PMID- 11914319 TI - Commentary: the polio model--does it apply for polio? PMID- 11914320 TI - Climatic, reservoir and occupational variables and the transmission of haemorrhagic fever with renal syndrome in China. AB - BACKGROUND: Being a zoonosis, the transmission of haemorrhagic fever with renal syndrome (HFRS) is influenced by climatic, reservoir and occupational factors. This paper has quantified the incidence and potential risk factors of HFRS in Yingshang County, a low-lying epidemic focus of the disease in China. METHODS: Correlation and regression analyses were conducted among climatic variables, the density of mice, autumn crop production and annual incidence of HFRS during the autumn-winter seasons in the County over the period 1980-1996. RESULTS: Rainfall (r = -0.63, P = 0.009), the density of mice (r = 0.90, P = 0.000) and autumn crop production (r = 0.67, P = 0.01) were statistically correlated with the incidence of HFRS. Multiple regression analysis indicated that these factors are potential predictors for HFRS transmission. CONCLUSIONS: Rainfall, the density of mice and autumn crop production could be used as predictors of HFRS transmission in low lying epidemic foci. PMID- 11914321 TI - Risk factors for trachoma in Mali. AB - OBJECTIVES: Prior to commencing a campaign to eliminate blinding trachoma in Mali, a national disease prevalence survey was conducted from March 1996 to June 1997. The prevalence of trachoma was estimated and potential risk factors were studied. METHODS: In each of Mali's seven regions (excluding the capital Bamako), a sample of 30 clusters was taken from the general population, in accordance with the principle of probability proportional to the size of the community. All children under 10 years of age were examined. The simplified clinical coding system proposed by the World Health Organization was used. The position of each village was established and subsequently related to the nearest meteorological station. Socioeconomic and environmental information was collected at both village and household level. The mother or caretaker of each child was questioned about availability and use of water for washing the child. At the time of examination, facial cleanliness and the presence of flies on the face were noted. RESULTS: A total of 15,187 children under 10 years of age were examined. The prevalence of active trachoma (follicular [TF] or intense trachoma [TI]) was 34.9% (95% CI : 32.3-37.6) and the prevalence of TI was 4.2% (95% CI : 3.5-5.0). Aridity/environmental dryness appears to be a risk factor influencing the current geographical distribution of trachoma. Small villages had considerably higher trachoma prevalence than their larger neighbours. The proximity of a medical centre and the existence of social organizations such as a women's association were associated with lower levels of trachoma. Crowded living conditions increased the risk. Using a monetary marker of wealth, we observed a linear inverse relation between wealth and trachoma prevalence. The presence of a dirty face was strongly associated with trachoma (odds ratio [OR] = 3.67) as was the presence of flies on the child's face (OR = 3.62). Trachoma prevalence increased with distance to a water source. Disease prevalence decreased with a higher frequency of both face washing and bathing. CONCLUSIONS: Of all the risk factors examined, facial cleanliness had the strongest association with the prevalence of trachoma. This was followed by the presence of flies on the child's face. Both face washing and bathing showed beneficial effects. Socioeconomic factors such as wealth were significantly explanatory. It is likely that hygiene education and fly control by environmental improvement could have a very significant impact on the prevalence of trachoma in Mali. PMID- 11914322 TI - Risk factors of invasive cervical cancer in Mali. AB - BACKGROUND: Cervical cancer is the most common cancer in women in Mali and the second commonest cause of cancer mortality. METHODS: As part of an international effort to evaluate the role of human papillomavirus (HPV) in the aetiology of cervical cancer, we conducted a hospital-based case-control study in three medical centres in Bamako during 1994-1995. A total of 82 cases (invasive cervical cancer patients) and 97 controls matched to the cases for age were included. Information on risk factors was collected through personal interview. Serum antibodies to HPV 16, 18 and 31 virus like particles (VLP) were detected using ELISA assays. Polymerase chain reaction was used to detect HPV DNA in frozen biopsies of cases. RESULTS: Human papillomavirus 6, 18, 31 VLP were detected in 60.4% of cases and 45.4% of controls (P = 0.03). Overall, HPV DNA was identified in 96.9% of the cervical cancer cases. Risk factors for cervical cancer were parity >10 versus <5 children ([odds ratio] OR = 4.8, 95% CI : 1.5 14.7), never having practised vaginal douching (OR = 17.6, 95% CI : 4.2-74.7), re using home-made feminine napkins (OR = 45.9, 95% CI : 8.8-238.7) and having a husband with more than two wives (OR = 5.3, 95% CI : 1.3-21.3). CONCLUSIONS: These data provide further evidence on the role of HPV in cervical cancer and show that high parity and poor genital hygiene conditions were the main co factors for cervical cancer in this population with prevalent HPV infection. PMID- 11914323 TI - Brain cancer and occupational exposure to magnetic fields among men: results from a Canadian population-based case-control study. AB - BACKGROUND: The relationship between occupational exposure to magnetic fields and brain cancer in men was investigated using population-based case-control data collected in eight Canadian provinces. Emphasis was placed on examining the variations in risk across different histological types. METHODS: A list of occupations was compiled for 543 cases and 543 controls that were individually matched by age. Occupations were categorized according to their average magnetic field exposure through blinded expert review (<0.3, 0.3-<0.6, and > or = 0.6 microT). In total, 133 cases (14%) and 123 controls (12%) were estimated to have at least one occupation whereby magnetic field exposures exceeded 0.3 microT. Odds ratios (OR) were generated using conditional logistic regression, and were adjusted for suspected occupational risk factors for brain cancer. RESULTS: A non significantly increased risk of brain cancer was observed among men who had ever held a job with an average magnetic field exposure >0.6 microT relative to those with exposures <0.3 microT (OR = 1.33, 95% CI : 0.75-2.36). A more pronounced risk was observed among men diagnosed with glioblastoma multiforme (OR = 5.36, 95% CI : 1.16-24.78). Moreover, a cumulative time weighted index score of magnetic field exposure was significantly related to glioblastoma multiforme (P = 0.02). In contrast, magnetic field exposures were not associated with astrocytoma or other brain cancers. CONCLUSIONS: Our findings support the hypothesis that occupational magnetic field exposure increases the risk of glioblastoma multiforme. PMID- 11914324 TI - Rural-to-urban migration and cardiovascular disease risk factors in young Guatemalan adults. AB - BACKGROUND: Migration to cities may increase cardiovascular disease risk factors in developing countries. We examined rural and urban individuals who were born in the same villages and shared similar childhood experiences. METHODS: Blood lipids and glucose, blood pressure, anthropometry, body composition, physical activity, and food, tobacco and alcohol consumption were examined in 161 men and 193 women, 19-29 years old, living in their village of birth (76 commuted to work in Guatemala City), and in 76 men and 43 women living in the city. RESULTS: Rural and urban women had similar prevalence of overweight (28%), elevated body fat (29.8 +/- 6.1%) and low physical activity (83%). Compared to rural men, more urban men were sedentary (79 versus 27%), and they had higher body fat (15.3 +/- 5.3% versus 13.3 +/- 5.7%), serum cholesterol (4.27 +/- 0.75 versus 3.90 +/- 0.70 mmol/l [165 +/- 29 versus 151 +/- 27 mg/dl]), low density lipoprotein [LDL] cholesterol (2.66 +/- 0.72 versus 2.30 +/- 0.62 mmol/l [103 +/- 28 versus 89 +/- 24 mg/dl]) and total cholesterol/high density lipoprotein [HDL]-cholesterol ratio (4.6 +/- 1.0 versus 4.1 +/- 0.9). Commuters showed intermediate values. Women had higher serum cholesterol (4.43 +/- 0.80 mmol/l [171 +/- 31 mg/dl]) than men in rural and urban areas. Urban residents ate/drank more saturated fats, red meat and sweetened beverages, and less legumes. CONCLUSIONS: High proportions of young Guatemalan women were overweight and sedentary. Migration to a city increased sedentarism and undesirable eating habits among men and women; men became fatter and their lipid profile worsened. Public health actions must address the prevention of emerging chronic diseases in countries still burdened by undernutrition and infections. PMID- 11914325 TI - Inverse relationship between urinary markers of animal protein intake and blood pressure in Chinese: results from the WHO Cardiovascular Diseases and Alimentary Comparison (CARDIAC) Study. AB - BACKGROUND: This issue of the relationship between animal protein intake and blood pressure (BP) is unsolved. We examined the associations between urinary 3 methylhistidine (3MH) excretion (a biological marker of animal protein intake) and BP in 11 Chinese population samples (Urumqi, Altai, Lhasa, Tulufan, Hetian, Guiyang, Guangzhou, Shanghai, Beijing, Shijiazhuang and Taipei). METHODS: This was a multi-centre cross-sectional study. In each centre, 100 men and 100 women aged 48-56 years were selected randomly from the general population. 3 methylhistidine in 24-hour (24-h) urine collections was measured by an Amino Acid Analyzer (Hitachi 835, Ibaragi, Japan). The total study sample included 966 men and 1025 women. Subjects who failed to collect complete 24-h urine samples were excluded in data analyses regarding associations between 3MH and BP. RESULTS: The results showed that: (1) for within-centre analyses of individuals, the 3MH and 3MH to creatinine ratios (3MH:cre) were significantly and negatively associated with BP and hypertension. These associations remained significant after adjustment for age, sex, sodium to potassium ratio, body mass index, calcium and magnesium. The pooled regression coefficients (SE) of systolic blood pressure (SBP) and diastolic blood pressure (DBP) on 3MH were -0.020 (0.01) and -0.018 (0.01), and of SBP and DBP on 3MH:cre were -0.022 (0.01) and -0.016 (0.01), respectively. Subjects with lower 3MH excretion had higher relative risks of hypertension than those who had higher 3MH excretion. (2) In cross-centre analyses, mean SBP and DBP were significantly and negatively associated with the mean 3MH:cre across the 11 population samples (R(2) = 0.56, P < 0.01). CONCLUSION: The results provide strong evidence that animal protein intake is associated inversely with BP in Chinese populations. PMID- 11914326 TI - Diabetes in a Caribbean population: epidemiological profile and implications. AB - OBJECTIVE: To examine the distribution and impact of diabetes, glycaemic status, and related factors, in a predominantly black adult Caribbean population. METHODS: The study included 4709 people, or 84% of a simple random sample of Barbadian-born citizens aged 40-84 years, examined between 1988 and 1992 and re assessed 4 years later. Diabetes was evaluated according to physician-diagnosis and glycosylated haemoglobin (GHb). Associations were assessed by logistic regression analyses, cumulative mortality by product-limit methods and death-rate ratios by Cox proportional hazards regression. RESULTS: Among the 4314 black participants, the prevalence of known diabetes, predominantly type 2, was 9.1% at 40-49 years of age and increased to 24.0% at 70-79 years. The overall prevalence was 17.5%, while it was 12.5% in mixed (black/ white; n = 184) and 6.0% in white/other participants (n = 133), only 0.3% had younger-onset. Additionally, 2% had GHb >10% (>2 SD over the mean) without diabetes history. Sulphonylureas were the most frequent treatment, while insulin use was infrequent. In black participants, diabetes was positively associated with age (OR = 1.03 per year; 95% CI : 1.02-1.04), diabetes family history (OR = 2.85, 95% CI : 2.39-3.40), hypertension (OR = 1.71, 95% CI : 1.42-2.05), obesity (BMI > or = 25 kg/m(2); OR = 1.74, 95% CI : 1.44-2.10), and high waist-hip ratio (WHR > or = 0.92; OR = 1.29, 95% CI : 1.09-1.53). Ocular co-morbidities were increased among people with diabetes, as was 4-year-mortality (death rate ratio = 1.42, 95% CI : 1.10-1.83). There was a 9% increase in mortality for each 1% increase in GHb (death rate ratio = 1.09, 95% CI : 1.04-1.15). CONCLUSIONS: A markedly high prevalence of diabetes existed in the adult black population, affecting almost one in five people and increasing morbidity and mortality. Prevention strategies are urgently needed to reduce the adverse implications of diabetes in this and similar populations. PMID- 11914327 TI - Distribution of blood pressure, body mass index and smoking habits in the urban population of Dar es Salaam, Tanzania, and associations with socioeconomic status. AB - OBJECTIVE: To estimate the distribution of blood pressure (BP), body mass index (BMI), smoking habits and their associations with socioeconomic status (SES) in an urban population in early epidemiological transition. METHODS: Cross-sectional survey of the entire population aged 25-64 years in five branches of Dar es Salaam (Tanzania) through visits to all homes in the study area. Blood pressure was based on the mean of the second and third readings with an automated device. Socioeconomic status was estimated with indicators of education, occupation and wealth. RESULTS: In all 9254 people were examined. Age-adjusted prevalence (%) among men/ women aged 35-64 years was 27.1/30.2 for BP > or = 140/90 mmHg or antihypertensive medication, 13.1/17.7 for BP > or = 160/95 mmHg or antihypertensive medication, 28.0/27.4 for BMI of 25.0-29.9 kg/m(2), 6.9/17.4 for BMI > or = 30 kg/m(2), and 22.0/2.6 for smoking (> or = 1 cigarette per day). Prevalence of categories of drinking frequency and history of diabetes are also reported. After adjustment for covariates, SES was associated inversely with BP and smoking and directly with BMI. Body mass index was associated positively with BP (1.01 and 0.91 mmHg systolic BP per 1 kg/m(2) BMI in men and women, respectively) and inversely with smoking (-1.14 kg/m(2) in male smokers versus non-smokers). Hypertension treatment rates were low, particularly in people of low SES. CONCLUSIONS: High prevalence of several cardiovascular risk factors in the urban population of a low-income country stresses the need for early public health interventions and adaptation of the health care infrastructure to meet the emerging challenge of cardiovascular disease. The direct SES-BMI association may drive increasing BMI and BP while the population becomes more affluent. PMID- 11914328 TI - Psychological distress as a risk factor for coronary heart disease in the Whitehall II Study. AB - BACKGROUND: Psychiatric disorder and psychological distress are increasingly recognized as risk factors for coronary heart disease (CHD). Elucidation of the mechanisms of these associations has implications for prevention. This study aims to confirm the association between psychological distress and CHD and examine if it could be explained by other factors such as health behaviours, social isolation and low control at work. METHODS: A prospective occupational cohort study of London-based civil service employees (Whitehall II Study) with baseline data collected from 1985-1988 with a 5-year follow-up. The participants were male and female middle-aged civil servants working in 20 Government Departments; 73% of eligible employees attended baseline screening. Psychological distress measured by the General Health Questionnaire (GHQ) at baseline was used to predict incidence of self-reported CHD and possible and probable electrocardiographic (ECG) abnormalities during follow-up. RESULTS: In men, baseline psychological distress was associated with an increased incidence of overall self-reported CHD (odds ratios [OR] = 1.83, 95% CI : 1.5-2.3) and ECG abnormalities (OR = 1.51, 95% CI : 1.1-2.1), after adjustment for age, employment grade and length of follow-up. In women, baseline psychological distress was also associated with an increased incidence of CHD (OR = 1.60, 95% CI : 1.2-2.1), but not with ECG abnormalities. Adjustment for health behaviours, marital status, social networks and work characteristics reduced the risks for incident CHD by 12% in men and by 10% in women; for ECG abnormalities these adjustments increased the risk in men by 16% and had little effect in women. CONCLUSIONS: The experience of psychological distress confers increased risk of CHD in men that is not explained by health behaviours, social isolation or work characteristics. The increased risk of CHD associated with psychological distress is not consistently demonstrated in women. PMID- 11914329 TI - Commentary: psychological distress: a matter of hearts and minds. PMID- 11914330 TI - Socioeconomic inequalities in all-cause and specific-cause mortality in Australia: 1985-1987 and 1995-1997. PMID- 11914333 TI - The world is ready to turn back the AIDS epidemic. PMID- 11914334 TI - AIDS epidemic faster in Eastern Europe than in rest of world. PMID- 11914335 TI - How Spiroplasma might swim. PMID- 11914336 TI - Thanks, Charley. PMID- 11914337 TI - Inferring genome trees by using a filter to eliminate phylogenetically discordant sequences and a distance matrix based on mean normalized BLASTP scores. AB - Darwin's paradigm holds that the diversity of present-day organisms has arisen via a process of genetic descent with modification, as on a bifurcating tree. Evidence is accumulating that genes are sometimes transferred not along lineages but rather across lineages. To the extent that this is so, Darwin's paradigm can apply only imperfectly to genomes, potentially complicating or perhaps undermining attempts to reconstruct historical relationships among genomes (i.e., a genome tree). Whether most genes in a genome have arisen via treelike (vertical) descent or by lateral transfer across lineages can be tested if enough complete genome sequences are used. We define a phylogenetically discordant sequence (PDS) as an open reading frame (ORF) that exhibits patterns of similarity relationships statistically distinguishable from those of most other ORFs in the same genome. PDSs represent between 6.0 and 16.8% (mean, 10.8%) of the analyzable ORFs in the genomes of 28 bacteria, eight archaea, and one eukaryote (Saccharomyces cerevisiae). In this study we developed and assessed a distance-based approach, based on mean pairwise sequence similarity, for generating genome trees. Exclusion of PDSs improved bootstrap support for basal nodes but altered few topological features, indicating that there is little systematic bias among PDSs. Many but not all features of the genome tree from which PDSs were excluded are consistent with the 16S rRNA tree. PMID- 11914338 TI - Anaerobic respiration using Fe(3+), S(0), and H(2) in the chemolithoautotrophic bacterium Acidithiobacillus ferrooxidans. AB - The chemolithoautotrophic bacterium Acidithiobacillus ferrooxidans has been known as an aerobe that respires on iron and sulfur. Here we show that the bacterium could chemolithoautotrophically grow not only on H(2)/O(2) under aerobic conditions but also on H(2)/Fe(3+), H(2)/S(0), or S(0)/Fe(3+) under anaerobic conditions. Anaerobic respiration using Fe(3+) or S(0) as an electron acceptor and H(2) or S(0) as an electron donor serves as a primary energy source of the bacterium. Anaerobic respiration based on reduction of Fe(3+) induced the bacterium to synthesize significant amounts of a c-type cytochrome that was purified as an acid-stable and soluble 28-kDa monomer. The purified cytochrome in the oxidized form was reduced in the presence of the crude extract, and the reduced cytochrome was reoxidized by Fe(3+). Respiration based on reduction of Fe(3+) coupled to oxidation of a c-type cytochrome may be involved in the primary mechanism of energy production in the bacterium on anaerobic iron respiration. PMID- 11914340 TI - Characterization of the Campylobacter jejuni heptosyltransferase II gene, waaF, provides genetic evidence that extracellular polysaccharide is lipid A core independent. AB - Campylobacter jejuni produces both lipooligosaccharide (LOS) and a higher molecular-weight polysaccharide that is believed to form a capsule. The role of these surface polysaccharides in C. jejuni-mediated enteric disease is unclear; however, epitopes associated with the LOS are linked to the development of neurological complications. In Escherichia coli and Salmonella enterica serovar Typhimurium the waaF gene encodes a heptosyltransferase, which catalyzes the transfer of the second L-glycero-D-manno-heptose residue to the core oligosaccharide moiety of lipopolysaccharide (LPS), and mutation of waaF results in a truncated core oligosaccharide. In this report we confirm experimentally that C. jejuni gene Cj1148 encodes the heptosyltransferase II enzyme, WaaF. The Campylobacter waaF gene complements an S. enterica serovar Typhimurium waaF mutation and restores the ability to produce full-sized lipopolysaccharide. To examine the role of WaaF in C. jejuni, waaF mutants were constructed in strains NCTC 11168 and NCTC 11828. Loss of heptosyltransferase activity resulted in the production of a truncated core oligosaccharide, failure to bind specific ligands, and loss of serum reactive GM(1), asialo-GM(1), and GM(2) ganglioside epitopes. The mutation of waaF did not affect the higher-molecular-weight polysaccharide supporting the production of a LOS-independent capsular polysaccharide by C. jejuni. The exact structural basis for the truncation of the core oligosaccharide was verified by comparative chemical analysis. The NCTC 11168 core oligosaccharide differs from that known for HS:2 strain CCUG 10936 in possessing an extra terminal disaccharide of galactose-beta(1,3) N-acetylgalactosamine. In comparison, the waaF mutant possessed a truncated molecule consistent with that observed with waaF mutants in other bacterial species. PMID- 11914339 TI - Xis protein binding to the left arm stimulates excision of conjugative transposon Tn916. AB - Tn916 and related conjugative transposons are clinically significant vectors for the transfer of antibiotic resistance among human pathogens, and they excise from their donor organisms using the transposon-encoded integrase ((Tn916)Int) and excisionase ((Tn916)Xis) proteins. In this study, we have investigated the role of the (Tn916)Xis protein in stimulating excisive recombination. The functional relevance of (Tn916)Xis binding sites on the arms of the transposon has been assessed in vivo using a transposon excision assay. Our results indicate that in Escherichia coli the stimulatory effect of the (Tn916)Xis protein is mediated by sequence-specific binding to either of its two binding sites on the left arm of the transposon. These sites lie in between the core and arm sites recognized by (Tn916)Int, suggesting that the (Tn916)Xis protein enhances excision in a manner similar to the excisionase protein of bacteriophage lambda, serving an architectural role in the stabilization of protein-nucleic acid structures required for strand synapsis. However, our finding that excision in E. coli is significantly enhanced by the host factor HU, but does not depend on the integration host factor or the factor for inversion stimulation, defines clear mechanistic differences between Tn916 and bacteriophage lambda recombination. PMID- 11914341 TI - Properties of the glucose-6-phosphate transporter from Chlamydia pneumoniae (HPTcp) and the glucose-6-phosphate sensor from Escherichia coli (UhpC). AB - The amino acid sequence of the proposed glucose-6-phosphate (Glc6P) transporter from Chlamydia pneumoniae (HPTcp; hexose phosphate transporter [Chlamydia pneumoniae]) exhibits a higher degree of similarity to the Escherichia coli Glc6P sensor (UhpC) than to the E. coli Glc6P transporter (UhpT). Overexpression of His UhpC in a UhpT-deficient E. coli strain revealed that the sensor protein is also able to transport Glc6P and exhibits an apparent K(m) ((Glc6P)) of 25 microM, whereas His-HPTcp exhibits an apparent K(m)( (Glc6P)) of 98 microM. His-HPTcp showed a four-times-lower specific activity than His-UhpT but a 56-times-higher specific activity than His-UhpC. Like His-UhpT and His-UhpC, the carrier His HPTcp performs a sugar-phosphate/inorganic-phosphate antiporter mode of transport. Surprisingly, while physiological concentrations of inorganic phosphate competitively inhibited transport mediated by the E. coli proteins His UhpT and His-UhpC, transport mediated by His-HPTcp was not inhibited. Interestingly, C(3)-organophosphates stimulated His-HPTcp activity but not His UhpT- or His-UhpC-catalyzed Glc6P transport. In contrast to His-UhpC, the His HPTcp protein does not act as a Glc6P sensor in the uhp regulon. PMID- 11914342 TI - Enterococcus faecalis acetoacetyl-coenzyme A thiolase/3-hydroxy-3-methylglutaryl coenzyme A reductase, a dual-function protein of isopentenyl diphosphate biosynthesis. AB - Many bacteria employ the nonmevalonate pathway for synthesis of isopentenyl diphosphate, the monomer unit for isoprenoid biosynthesis. However, gram-positive cocci exclusively use the mevalonate pathway, which is essential for their growth (E. I. Wilding et al., J. Bacteriol. 182:4319-4327, 2000). Enzymes of the mevalonate pathway are thus potential targets for drug intervention. Uniquely, the enterococci possess a single open reading frame, mvaE, that appears to encode two enzymes of the mevalonate pathway, acetoacetyl-coenzyme A thiolase and 3 hydroxy-3-methylglutaryl-coenzyme A (HMG-CoA) reductase. Western blotting revealed that the mvaE gene product is a single polypeptide in Enterococcus faecalis, Enterococcus faecium, and Enterococcus hirae. The mvaE gene was cloned from E. faecalis and was expressed with an N-terminal His tag in Escherichia coli. The gene product was then purified by nickel affinity chromatography. As predicted, the 86.5-kDa mvaE gene product catalyzed both the acetoacetyl-CoA thiolase and HMG-CoA reductase reactions. Temperature optima, DeltaH(a) and K(m) values, and pH optima were determined for both activities. Kinetic studies of acetoacetyl-CoA thiolase implicated a ping-pong mechanism. CoA acted as an inhibitor competitive with acetyl-CoA. A millimolar K(i) for a statin drug confirmed that E. faecalis HMG-CoA reductase is a class II enzyme. The oxidoreductant was NADP(H). A role for an active-site histidine during the first redox step of the HMG-CoA, reductase reaction was suggested by the ability of diethylpyrocarbonate to block formation of mevalonate from HMG-CoA, but not from mevaldehyde. Sequence comparisons with other HMG-CoA reductases suggest that the essential active-site histidine is His756. The mvaE gene product represents the first example of an HMG-CoA reductase fused to another enzyme. PMID- 11914343 TI - Population genetic structure of Legionella pneumophila inferred from RNA polymerase gene (rpoB) and DotA gene (dotA) sequences. AB - The population structure of Legionella pneumophila was studied by using partial RNA polymerase gene (rpoB) and DotA gene (dotA) sequences. Trees inferred from rpoB sequences showed that two subspecies of L. pneumophila, Legionella pneumophila subsp. pneumophila and Legionella pneumophila subsp. fraseri, were clearly separated genetically. In both rpoB and dotA trees, 79 Korean isolates used in this study constituted six clonal populations, four of which (designated subgroups P-I to P-IV) were identified in L. pneumophila subsp. pneumophila and two of which (designated subgroups F-I and F-II) were identified in L. pneumophila subsp. fraseri. Although the relationships among subgroups were not identical, such subgrouping was congruent between the rpoB and dotA trees. Type strains of several serogroups did not belong to any subgroup, presumably because isolates similar to these strains were not present among our local sample of the population. There was evidence that horizontal gene transfer or recombination had occurred within L. pneumophila. Contrary to the phylogeny from rpoB and the taxonomic context, subgroups P-III and P-IV of L. pneumophila subsp. pneumophila proved to be closely related to those of L. pneumophila subsp. fraseri or showed a distinct clustering in the dotA tree. It can be inferred that dotA of subgroups P-III and P-IV has been transferred horizontally from other subspecies. The diverse distribution of serogroup 1 strains through the gene trees suggests that surface antigen-coding genes that determine serogroup can be exchanged. Thus, it can be inferred that genetic recombination has been important in the evolution of L. pneumophila. PMID- 11914344 TI - Mutations in the 16S rRNA genes of Helicobacter pylori mediate resistance to tetracycline. AB - Low-cost and rescue treatments for Helicobacter pylori infections involve combinations of several drugs including tetracycline. Resistance to tetracycline has recently emerged in H. pylori. The 16S rRNA gene sequences of two tetracycline-resistant clinical isolates (MIC = 64 microg/ml) were determined and compared to the consensus H. pylori 16S rRNA sequence. One isolate had four nucleotide substitutions, and the other had four substitutions and two deletions. Natural transformation with the 16S rRNA genes from the resistant organisms conferred tetracycline resistance on susceptible strains. 16S rRNA genes containing the individual mutations were constructed and tested for the ability to confer resistance. Only the 16S rRNA gene containing the triple mutation, AGA965-967TTC, was able to confer tetracycline resistance on H. pylori 26695. The MICs of tetracycline for the transformed strains were equivalent to those for the original clinical isolates. The two original isolates were also metronidazole resistant, but this trait was not linked to the tetracycline resistance phenotype. Serial passage of several H. pylori strains on increasing concentrations of tetracycline yielded mutants with only a very modest increase in tetracycline resistance to a MIC of 4 to 8 microg/ml. These mutants all had a deletion of G942 in the 16S rRNA genes. The mutations in the 16S rRNA are clearly responsible for tetracycline resistance in H. pylori. PMID- 11914345 TI - Further evidence that a cell wall precursor [C(55)-MurNAc-(peptide)-GlcNAc] serves as an acceptor in a sorting reaction. AB - Previous studies suggested that a Gly-containing branch of cell wall precursor [C(55)-MurNAc-(peptide)-GlcNAc], which is often referred to as lipid II, might serve as a nucleophilic acceptor in sortase-catalyzed anchoring of surface proteins in Staphylococcus aureus. To test this hypothesis, we first simplified the procedure for in vitro biosynthesis of Gly-containing lipid II by using branched UDP-MurNAc-hexapeptide isolated from the cytoplasm of Streptomyces spp. Second, we designed a thin-layer chromatography-based assay in which the mobility of branched but not linear lipid II is shifted in the presence of both sortase and LPSTG-containing peptide. These results and those of additional experiments presented in this study further suggest that lipid II indeed serves as a natural substrate in a sorting reaction. PMID- 11914346 TI - Bacillus subtilis 168 contains two differentially regulated genes encoding L asparaginase. AB - Expression of the two Bacillus subtilis genes encoding L-asparaginase is controlled by independent regulatory factors. The ansZ gene (formerly yccC) was shown by mutational analysis to encode a functional L-asparaginase, the expression of which is activated during nitrogen-limited growth by the TnrA transcription factor. Gel mobility shift and DNase I footprinting experiments indicate that TnrA regulates ansZ expression by binding to a DNA site located upstream of the ansZ promoter. The expression of the ansA gene, which encodes the second L-asparaginase, was found to be induced by asparagine. The ansA repressor, AnsR, was shown to negatively regulate its own expression. PMID- 11914347 TI - Use of the Caulobacter crescentus genome sequence to develop a method for systematic genetic mapping. AB - The functional analysis of sequenced genomes will be facilitated by the development of tools for the rapid mapping of mutations. We have developed a systematic approach to genetic mapping in Caulobacter crescentus that is based on bacteriophage-mediated transduction of strategically placed antibiotic resistance markers. The genomic DNA sequence was used to identify sites distributed evenly around the chromosome at which plasmids could be nondisruptively integrated. DNA fragments from these sites were amplified by PCR and cloned into a kanamycin resistant (Kan(r)) suicide vector. Delivery of these plasmids into C. crescentus resulted in integration via homologous recombination. A set of 41 strains containing Kan(r) markers at 100-kb intervals was thereby generated. These strains serve as donors for generalized transduction using bacteriophage phiCr30, which can transduce at least 120 kb of DNA. Transductants are selected with kanamycin and screened for loss of the mutant phenotype to assess linkage between the marker and the site of the mutation. The dependence of cotransduction frequency on sequence distance was evaluated using several markers and mutant strains. With these data as a standard, previously unmapped mutations were readily localized to DNA sequence intervals equivalent to less than 1% of the genome. Candidate genes within the interval were then examined further by subcloning and complementation analysis. Mutations resulting in sensitivity to ampicillin, in nutritional auxotrophies, or temperature-sensitive growth were mapped. This approach to genetic mapping should be applicable to other bacteria with sequenced genomes for which generalized transducing phage are available. PMID- 11914348 TI - Effects of pyrazinamide on fatty acid synthesis by whole mycobacterial cells and purified fatty acid synthase I. AB - The effects of low extracellular pH and intracellular accumulation of weak organic acids were compared with respect to fatty acid synthesis by whole cells of Mycobacterium tuberculosis and Mycobacterium smegmatis. The profile of fatty acids synthesized during exposure to benzoic, nicotinic, or pyrazinoic acids, as well as that observed during intracellular hydrolysis of the corresponding amides, was not a direct consequence of modulation of fatty acid synthesis by these compounds but reflected the response to inorganic acid stress. Analysis of fatty acid synthesis in crude mycobacterial cell extracts demonstrated that pyrazinoic acid failed to directly modulate the fatty acid synthase activity catalyzed by fatty acid synthase I (FAS-I). However, fatty acid synthesis was irreversibly inhibited by 5-chloro-pyrazinamide in a time-dependent fashion. Moreover, we demonstrate that pyrazinoic acid does not inhibit purified mycobacterial FAS-I, suggesting that this enzyme is not the immediate target of pyrazinamide. PMID- 11914349 TI - Functional and mutational analysis of conjugative transfer region 1 (Tra1) from the IncHI1 plasmid R27. AB - The conjugative transfer region 1 (Tra1) of the IncHI1 plasmid R27 was subjected to DNA sequence analysis, mutagenesis, genetic complementation, and an H-pilus specific phage assay. Analysis of the nucleotide sequence indicated that the Tra1 region contains genes coding for mating pair formation (Mpf) and DNA transfer replication (Dtr) and a coupling protein. Insertional disruptions of 9 of the 14 open reading frames (ORFs) in the Tra1 region resulted in a transfer-deficient phenotype. Conjugative transfer was restored for each transfer mutant by genetic complementation. An intergenic region between traH and trhR was cloned and mobilized by R27, indicating the presence of an origin of transfer (oriT). The five ORFs immediately downstream of the oriT region are involved in H-pilus production, as determined by an H-pilus-specific phage assay. Three of these ORFs encode proteins homologous to Mpf proteins from IncF plasmids. Upstream of the oriT region are four ORFs required for plasmid transfer but not H-pilus production. TraI contains sequence motifs that are characteristic of relaxases from the IncP lineage but share no overall homology to known relaxases. TraJ contains both an Arc repressor motif and a leucine zipper motif. A putative coupling protein, TraG, shares a low level of homology to the TraG family of coupling proteins and contains motifs that are important for DNA transfer. This analysis indicates that the Mpf components of R27 share a common lineage with those of the IncF transfer system, whereas the relaxase of R27 is ancestrally related to that of the IncP transfer system. PMID- 11914350 TI - Differential recognition of surface proteins in Streptococcus pyogenes by two sortase gene homologs. AB - The interaction of Streptococcus pyogenes (group A streptococcus [GAS]) with its human host requires several surface proteins. In this study, we isolated mutations in a gene required for the surface localization of protein F by transposon mutagenesis of the M6 strain JRS4. This gene (srtA) encodes a protein homologous to Staphylococcus aureus sortase, which covalently links proteins containing an LPXTG motif to the cell wall. The GAS srtA mutant was defective in anchoring the LPXTG-containing proteins M6, protein F, ScpA, and GRAB to the cell surface. This phenotype was complemented when a wild-type srtA gene was provided in trans. The surface localization of T6, however, was unaffected by the srtA mutation. The M1 genome sequence contains a second open reading frame with a motif characteristic of sortase proteins. Inactivation of this gene (designated srtB) in strain JRS4 affected the surface localization of T6 but not M6, protein F, ScpA, or GRAB. This phenotype was complemented by srtB in trans. An srtA probe hybridized with DNA from all GAS strains tested (M types 1, 3, 4, 5, 6, 18, 22, and 50 and nontypeable strain 64/14) and from streptococcal groups C and G, while srtB hybridized with DNA from only a few GAS strains. We conclude that srtA and srtB encode sortase enzymes required for anchoring different subsets of proteins to the cell wall. It seems likely that the multiple sortase homologs in the genomes of other gram-positive bacteria have a similar substrate-specific role. PMID- 11914351 TI - Expression, autoregulation, and DNA binding properties of the Mycobacterium tuberculosis TrcR response regulator. AB - The TrcRS two-component system of Mycobacterium tuberculosis is comprised of the TrcS histidine kinase and the TrcR response regulator, which is homologous to the OmpR class of DNA binding response regulators. Reverse transcription-PCRs with total RNA showed that the trcR and trcS two-component system genes are transcribed in broth-grown M. tuberculosis. Analysis of the trcR and trcS genes using various SCOTS (selective capture of transcribed sequences) probes also confirmed that these genes are expressed in broth-grown cultures and after 18 h of M. tuberculosis growth in cultured human primary macrophages. To determine if the TrcR response regulator is autoregulated, a trcR-lacZ fusion plasmid and a TrcR expression plasmid were cotransformed into Escherichia coli. Upon induction of the TrcR protein, there was a >500-fold increase in beta-galactosidase activity from the trcR-lacZ fusion, indicating that TrcR is involved in transcriptional autoactivation. Gel mobility shift assays with the trcR promoter and TrcR established that the response regulator was autoregulating via direct binding. By use of a delimiting series of overlapping trcR PCR fragments in gel mobility shift assays with TrcR, an AT-rich region of the trcR promoter was shown to be essential for TrcR binding. Additionally, this AT-rich sequence was protected by TrcR in DNase I protection assays. To further analyze the role of the AT-rich region in TrcR autoregulation, the trcR promoter was mutated and analyzed in lacZ transcriptional fusions in the presence of TrcR. Alteration of the AT-rich sequence in the trcR promoter resulted in the loss of trcR transcriptional activation in the presence of TrcR. This report indicates that the M. tuberculosis TrcR response regulator activates its own expression by interacting with the AT-rich sequence of the trcR promoter. PMID- 11914352 TI - Origin binding activity of the Mycobacterial plasmid pAL5000 replication protein RepB is stimulated through interactions with host factors and coupled expression of repA. AB - The minimal replication region of the mycobacterial plasmid pAL5000 encompasses the replication origin (ori) and two tandemly organized replication genes, repA and repB, the functions of which are not clearly known. It was observed that when the repA and repB genes were expressed in Escherichia coli, a strong ori binding activity was generated in the host cells. Inactivation of repB led to a complete loss of activity, whereas inactivation of repA had a partial effect, indicating that while repB plays an important role in the process, its activity is stimulated through coexpression of repA. However, this stimulatory effect could be demonstrated only when expression of repA and that of repB were coupled. At a relatively high concentration (1,000 nM), the purified RepB protein was found to form an ori complex with low specificity, which was sensitive to high salt concentrations and challenge by a nonspecific competitor. In contrast, the complex formed by an extract of repA-repB-expressing cells was highly specific and was resistant to both types of challenges. At a 10-fold-lower concentration, RepB did not exhibit ori binding activity, but it could nevertheless form a salt resistant ori complex in vitro, provided that host factors were present. Antibody supershift experiments indicated that RepB is a key component of the specific complex formed by extracts prepared from E. coli cells expressing the repA and repB genes and also from mycobacterial cells harboring pAL5000-derived vectors. The results indicate that in vivo RepB interacts with host factors and forms an ori complex, but this activity is maximal only when there is coupled expression of repA. PMID- 11914353 TI - Role for vitamin B(12) in light induction of gene expression in the bacterium Myxococcus xanthus. AB - A light-inducible promoter (P(B)) drives the carB operon (carotenoid genes) of the bacterium Myxococcus xanthus. A gene encoding a regulator of carotenoid biosynthesis was identified by studying mutant strains carrying a transcriptional fusion to P(B) and deletions in three candidate genes. Our results prove that the identified gene, named carA, codes for a repressor of the P(B) promoter in the dark. They also show that the carA gene product does not participate in the light activation of two other promoters connected with carotenoid synthesis or its regulation in M. xanthus. CarA is a novel protein consisting of a DNA-binding domain of the family of MerR helix-turn-helix transcriptional regulators, directly joined to a cobalamin-binding domain. In support of this, we report here that the presence of vitamin B(12) or some other cobalamin derivatives is absolutely required for activation of the P(B) promoter by light. PMID- 11914354 TI - The extracellular transport signal of the Vibrio cholerae endochitinase (ChiA) is a structural motif located between amino acids 75 and 555. AB - ChiA, an 88-kDa endochitinase encoded by the chiA gene of the gram-negative enteropathogen Vibrio cholerae, is secreted via the eps-encoded main terminal branch of the general secretory pathway (GSP), a mechanism which also transports cholera toxin. To localize the extracellular transport signal of ChiA that initiates transport of the protein through the GSP, a chimera comprised of ChiA fused at the N terminus with the maltose-binding protein (MalE) of Escherichia coli and fused at the C terminus with a 13-amino-acid epitope tag (E-tag) was expressed in strain 569B(chiA::Kan(r)), a chiA-deficient but secretion-competent mutant of V. cholerae. Fractionation studies revealed that blockage of the natural N terminus and C terminus of ChiA did not prevent secretion of the MalE ChiA-E-tag chimera. To locate the amino acid sequences which encoded the transport signal, a series of truncations of ChiA were engineered. Secretion of the mutant polypeptides was curtailed only when ChiA was deleted from the N terminus beyond amino acid position 75 or from the C terminus beyond amino acid 555. A mutant ChiA comprised of only those amino acids was secreted by wild-type V. cholerae but not by an epsD mutant, establishing that amino acids 75 to 555 independently harbored sufficient structural information to promote secretion by the GSP of V. cholerae. Cys77 and Cys537, two cysteines located just within the termini of ChiA(75-555), were not required for secretion, indicating that those residues were not essential for maintaining the functional activity of the ChiA extracellular transport signal. PMID- 11914355 TI - Gene transfer between Salmonella enterica serovar Typhimurium inside epithelial cells. AB - Virulence and antibiotic resistance genes transfer between bacteria by bacterial conjugation. Conjugation also mediates gene transfer from bacteria to eukaryotic organisms, including yeast and human cells. Predicting when and where genes transfer by conjugation could enhance our understanding of the risks involved in the release of genetically modified organisms, including those being developed for use as vaccines. We report here that Salmonella enterica serovar Typhimurium conjugated inside cultured human cells. The DNA transfer from donor to recipient bacteria was proportional to the probability that the two types of bacteria occupied the same cell, which was dependent on viable and invasive bacteria and on plasmid tra genes. Based on the high frequencies of gene transfer between bacteria inside human cells, we suggest that such gene transfers occur in situ. The implications of gene transfer between bacteria inside human cells, particularly in the context of antibiotic resistance, are discussed. PMID- 11914356 TI - Roles of the C-terminal end of SecY in protein translocation and viability of Escherichia coli. AB - SecY, a central component of the membrane-embedded sector of protein translocase, contains six cytosolic domains. Here, we examined the importance of the C terminal cytosolic region of SecY by systematically shortening the C-terminal end and examining the functional consequences of these mutations in vivo and in vitro. It was indicated that the C-terminal five residues are dispensable without any appreciable functional defects in SecY. Mutants missing the C-terminal six to seven residues were partially compromised, especially at low temperature or in the absence of SecG. In vitro analyses indicated that the initial phase of the translocation reaction, in which the signal sequence region of the preprotein is inserted into the membrane, was affected by the lack of the C-terminal residues. SecA binding was normal, but SecA insertion in response to ATP and a preprotein was impaired. It is suggested that the C-terminal SecY residues are required for SecA-dependent translocation initiation. PMID- 11914357 TI - Distinct roles of P(II)-like signal transmitter proteins and amtB in regulation of nif gene expression, nitrogenase activity, and posttranslational modification of NifH in Azoarcus sp. strain BH72. AB - P(II)-like signal transmitter proteins, found in Bacteria, Archaea, and plants, are known to mediate control of carbon and nitrogen assimilation. They indirectly regulate the activity of key metabolic enzymes and transcription factors by protein-protein interactions with signal transduction proteins. Many Proteobacteria harbor two paralogous P(II)-like proteins, GlnB and GlnK, whereas a novel third P(II) paralogue (GlnY) was recently identified in Azoarcus sp. strain BH72, a diazotrophic endophyte of grasses. In the present study, evidence was obtained that the P(II)-like proteins have distinct roles in mediating nitrogen and oxygen control of nif gene transcription and nitrogenase activity. Full repression of nif gene transcription in the presence of a combined nitrogen source or high oxygen concentrations was observed in wild-type and glnB and glnK knockout mutants, revealing that GlnB and GlnK can complement each other in mediating the repression. In contrast, in a glnBK double mutant strain in the presence of only GlnY, nif gene transcription was still detectable, albeit at a lower level, on nitrate or 20% oxygen. As another level of control, nitrogenase activity was regulated by at least three types of mechanisms in strain BH72: covalent modification of dinitrogenase reductase (NifH), probably by ADP ribosylation, and two other, unknown means. Functional inactivation upon ammonium addition (switch-off) required the putative high-affinity ammonium transporter AmtB and GlnK, but not GlnB or GlnY. Functional inactivation in response to anaerobiosis did not depend on AmtB, GlnK, or GlnB. In contrast, covalent modification of NifH required both GlnB and GlnK and AmtB as response to ammonium addition, whereas it required either GlnB or GlnK and not AmtB when cells were shifted to anaerobiosis. In a glnBK double mutant expressing only GlnY, NifH modification was completely abolished, further revealing functional differences between the three P(II) paralogues. PMID- 11914359 TI - Determinants of the C-terminal domain of the Escherichia coli RNA polymerase alpha subunit important for transcription at class I cyclic AMP receptor protein dependent promoters. AB - Alanine scanning of the Escherichia coli RNA polymerase alpha subunit C-terminal domain (alphaCTD) was used to identify amino acid side chains important for class I cyclic AMP receptor protein (CRP)-dependent transcription. Key residues were investigated further in vivo and in vitro. Substitutions in three regions of alphaCTD affected class I CRP-dependent transcription from the CC(-61.5) promoter and/or the lacP1 promoter. These regions are (i) the 287 determinant, previously shown to contact CRP during class II CRP-dependent transcription; (ii) the 265 determinant, previously shown to be important for alphaCTD-DNA interactions, including those required for class II CRP-dependent transcription; and (iii) the 261 determinant. We conclude that CRP contacts the same target in alphaCTD, the 287 determinant, at class I and class II CRP-dependent promoters. We also conclude that the relative contributions of individual residues within the 265 determinant depend on promoter sequence, and we discuss explanations for effects of substitutions in the 261 determinant. PMID- 11914360 TI - Global regulation by gidA in Pseudomonas syringae. AB - Analysis of two virulence mutants of Pseudomonas syringae B728a revealed that the Tn 5 sites of insertion were within the gidA open reading frame (ORF). These mutations were pleiotropic, affecting diverse phenotypic traits, such as lipodepsipeptide (syringomycin and syringopeptin) antibiotic production, swarming, presence of fluorescent pigment, and virulence. Site-specific recombination of a disrupted gidA gene into the chromosome resulted in the same phenotypic pattern as transposon insertion. Mutant phenotypes were restored by the gidA ORF on a plasmid. The salA gene, a copy number suppressor of the syringomycin-deficient phenotype in gacS and gacA mutants, was also found to suppress the antibiotic-negative phenotypes of gidA mutants, suggesting that gidA might play some role in salA regulation. Reporter studies with chromosomal salA lacZ translational fusions confirmed that salA reporter expression decreased approximately fivefold in a gidA mutant background, with a concurrent decrease in the expression of the syringomycin biosynthetic reporter fusion syrB-lacZ. Wild type levels of reporter expression were restored by supplying an intact gidA gene on a plasmid. Often described as being involved in cell division, more recent evidence suggests a role for gidA in moderating translational fidelity, suggesting a mechanism by which global regulation might occur. The gidA gene is essentially universal in the domains Bacteria and Eucarya but has no counterparts in Archaea, probably reflecting specific differences in the translational machinery between the former and latter domains. PMID- 11914358 TI - Evolutionary analysis by whole-genome comparisons. AB - A total of 37 complete genome sequences of bacteria, archaea, and eukaryotes were compared. The percentage of orthologous genes of each species contained within any of the other 36 genomes was established. In addition, the mean identity of the orthologs was calculated. Several conclusions result: (i) a greater absolute number of orthologs of a given species is found in larger species than in smaller ones; (ii) a greater percentage of the orthologous genes of smaller genomes is contained in other species than is the case for larger genomes, which corresponds to a larger proportion of essential genes; (iii) before species can be specifically related to one another in terms of gene content, it is first necessary to correct for the size of the genome; (iv) eukaryotes have a significantly smaller percentage of bacterial orthologs after correction for genome size, which is consistent with their placement in a separate domain; (v) the archaebacteria are specifically related to one another but are not significantly different in gene content from the bacteria as a whole; (vi) determination of the mean identity of all orthologs (involving hundreds of gene comparisons per genome pair) reduces the impact of errors in misidentification of orthologs and to misalignments, and thus it is far more reliable than single gene comparisons; (vii) however, there is a maximum amount of change in protein sequences of 37% mean identity, which limits the use of percentage sequence identity to the lower taxa, a result which should also be true for single gene comparisons of both proteins and rRNA; (viii) most of the species that appear to be specifically related based upon gene content also appear to be specifically related based upon the mean identity of orthologs; (ix) the genes of a majority of species considered in this study have diverged too much to allow the construction of all-encompassing evolutionary trees. However, we have shown that eight species of gram-negative bacteria, six species of gram-positive bacteria, and eight species of archaebacteria are specifically related in terms of gene content, mean identity of orthologs, or both. PMID- 11914361 TI - AniA regulates reserve polymer accumulation and global protein expression in Rhizobium etli. AB - Previously, it was reported that the oxidative capacity and ability to grow on carbon sources such as pyruvate and glucose were severely diminished in the Rhizobium etli phaC::OmegaSm(r)/Sp(r) mutant CAR1, which is unable to synthesize poly-beta-hydroxybutyric acid (PHB) (M. A. Cevallos, S. Encarnacion, A. Leija, Y. Mora, and J. Mora, J. Bacteriol. 178:1646-1654, 1996). By random Tn5 mutagenesis of the phaC strain, we isolated the mutants VEM57 and VEM58, both of which contained single Tn5 insertions and had recovered the ability to grow on pyruvate or glucose. Nucleotide sequencing of the region surrounding the Tn5 insertions showed that they had interrupted an open reading frame designated aniA based on its high deduced amino acid sequence identity to the aniA gene product of Sinorhizobium meliloti. R. etli aniA was located adjacent to and divergently transcribed from genes encoding the PHB biosynthetic enzymes beta-ketothiolase (PhaA) and acetoacetyl coenzyme A reductase (PhaB). An aniA::Tn5 mutant (VEM5854) was constructed and found to synthesize only 40% of the wild type level of PHB. Both VEM58 and VEM5854 produced significantly more extracellular polysaccharide than the wild type. Organic acid excretion and levels of intracellular reduced nucleotides were lowered to wild-type levels in VEM58 and VEM5854, in contrast to those of strain CAR1, which were significantly elevated. Proteome analysis of VEM58 showed a drastic alteration of protein expression, including the absence of a protein identified as PhaB. We propose that the aniA gene product plays an important role in directing carbon flow in R. etli. PMID- 11914362 TI - Effect of aniA (carbon flux regulator) and PhaC (poly-beta-hydroxybutyrate synthase) mutations on pyruvate metabolism in Rhizobium etli. AB - The Rhizobium etli poly-beta-hydroxybutyrate synthase (PhaC) mutant SAM100 grows poorly with pyruvate as the carbon source. The inactivation of aniA, encoding a global carbon flux regulator, in SAM100 restores growth of the resulting double mutant (VEM58) on pyruvate. Pyruvate carboxylase (PYC) activity, pyc gene transcription, and holoenzyme content, which were low in SAM100, were restored in strain VEM58. The genetically engineered overexpression of PYC in SAM100 also allowed its growth on pyruvate. The possible relation between AniA, pyc transcription, and reduced-nucleotide levels is discussed. PMID- 11914363 TI - The crystal structure of Zn(II)-free Treponema pallidum TroA, a periplasmic metal binding protein, reveals a closed conformation. AB - We previously demonstrated that Treponema pallidum TroA is a periplasmic metal binding protein (MBP) with a distinctive alpha-helical backbone. To better understand the mechanisms of metal binding and release by TroA, we determined the crystal structure of the apoprotein at a resolution of 2.5 A and compared it to that of the Zn(II)-bound form (Protein Data Bank accession code 1toa). apo-TroA shows a conformation even more closed than that of its Zn(II)-bound counterpart due to a 4 degrees tilt of the C-terminal domain (residues 190 through 308) about an axis parallel to the poorly flexible backbone helix. This domain tilting pushes two loops (residues 248 through 253 and 277 through 286) towards the metal binding site by more than 1 A, resulting in an unfavorable interaction of I251 with D66. To avoid this contact, D66 shifts towards H68, one of the four Zn(II) coordinating residues. The approach of this negative charge coincides with the flipping of the imidazole side chain of H68, resulting in the formation of a new hydrogen bond. The conformational change of H68, along with a slight rearrangement of D279, a C-terminal domain Zn(II)-coordinating residue, distorts the metal-binding site geometry, presumably causing the release of the bound metal ion. Ligand binding and release by TroA, and presumably by other members of the MBP cluster, differs from the "Venus flytrap" mechanism utilized by bacterial nonmetal solute-binding receptors. PMID- 11914364 TI - Type 4 pilus biogenesis and type II-mediated protein secretion by Vibrio cholerae occur independently of the TonB-facilitated proton motive force. AB - In Vibrio cholerae, elaboration of toxin-coregulated pilus and protein secretion by the extracellular protein secretion apparatus occurred in the absence of both TonB systems. In contrast, the cognate putative ATPases were required for each process and could not substitute for each other. PMID- 11914365 TI - A MecA paralog, YpbH, binds ClpC, affecting both competence and sporulation. AB - ComK, the master regulator of competence, is degraded by the general stress related protease ClpCP but must be targeted to this protease by binding to the adapter protein MecA. The genome of Bacillus subtilis contains a paralog of mecA, ypbH. We show in the present study that YpbH, like MecA, binds ClpC and that its elimination or overproduction affects competence and sporulation. PMID- 11914366 TI - Prediction of gene function in methylthioadenosine recycling from regulatory signals. AB - The S-box transcription termination control system, first identified in Bacillus subtilis, is used for regulation of gene expression in response to methionine availability. The presence of the S-box motif provided the first indication that the ykrTS and ykrWXYZ genes could play a role in recycling of 5' methylthioadenosine, a by-product of polyamine biosynthesis that can be converted to methionine. In this study we demonstrate a role for the ykrTS and ykrWXYZ gene products in this pathway. PMID- 11914367 TI - EvgA of the two-component signal transduction system modulates production of the yhiUV multidrug transporter in Escherichia coli. AB - Overexpression of the EvgA regulator of the two-component signal transduction system was previously found to modulate multidrug resistance of Escherichia coli by increasing efflux of drugs (K. Nishino and A. Yamaguchi, J. Bacteriol. 183:1455-1458, 2001). Here we present data showing that EvgA contributes to multidrug resistance through increased expression of the multidrug transporter yhiUV gene. PMID- 11914368 TI - TetR family member psrA directly binds the Pseudomonas rpoS and psrA promoters. AB - We have previously described a Pseudomonas gene, psrA, which enhances transcription of the rpoS sigma factor gene at stationary phase. We present molecular data which demonstrate that in Pseudomonas putida PsrA binds specifically to the rpoS and psrA promoters in DNA regions having similar palindromic sequences, C/GAAAC N(2-4) GTTTG/C, where N is any nucleotide. The position of the initiation of transcription was determined for both promoters, and PsrA binds from positions -59 to -35 in the rpoS promoter and from -18 to +20 in the psrA promoter with respect to the +1 transcription site. Expression studies with a psrA-lacZ transcriptional fusion in wild-type and psrA::Tn5 knockout mutants revealed that psrA was under additional control in response to growth phase. A model for the role of PsrA in the regulation of rpoS and psrA is presented. PMID- 11914369 TI - The WNT7b promoter is regulated by TTF-1, GATA6, and Foxa2 in lung epithelium. AB - In this study, we find that WNT7b is the only member of the WNT family of autocrine/paracrine signaling molecules whose expression in the lung is restricted to the airway epithelium during embryonic development. To study the transcriptional mechanisms that underlie this restricted pattern of WNT7b expression, we isolated the proximal 1.0-kb mouse WNT7b promoter and mapped the transcriptional start sites. Transfection of the lung epithelial cell line MLE 15, which expresses WNT7b, shows that the 1.0-kb mouse WNT7b promoter is highly active in lung epithelial cells. This region of the WNT7b promoter contains several DNA binding sites for the important lung-restricted transcription factors TTF-1, GATA6, and Foxa2. Electrophoretic mobility shift assays showed that TTF-1, GATA6, and Foxa2 can bind to a specific subset of their consensus DNA binding sites within the WNT7b promoter. Using cotransfection assays, we demonstrate that TTF-1, GATA6, and Foxa2 can trans-activate the WNT7b promoter in NIH-3T3 cells. Truncation of GATA6 or Foxa2 binding sites reduced the ability of these transcriptional regulators to trans-activate the WNT7b promoter. Finally, the minimal 118-bp region of the mouse WNT7b promoter containing only TTF-1 binding sites was synergistically activated by TTF-1 and GATA6, and we show that TTF-1 and GATA6 physically interact in vivo. Together, these results suggest that WNT7b gene expression in the lung epithelium is regulated in a combinatorial fashion by TTF-1, GATA6, and Foxa2. PMID- 11914371 TI - The crystal structure of Mycobacterium tuberculosis alkylhydroperoxidase AhpD, a potential target for antitubercular drug design. AB - The resistance of Mycobacterium tuberculosis to isoniazid is commonly linked to inactivation of a catalase-peroxidase, KatG, that converts isoniazid to its biologically active form. Loss of KatG is associated with elevated expression of the alkylhydroperoxidases AhpC and AhpD. AhpD has no sequence identity with AhpC or other proteins but has alkylhydroperoxidase activity and possibly additional physiological activities. The alkylhydroperoxidase activity, in the absence of KatG, provides an important antioxidant defense. We have determined the M. tuberculosis AhpD structure to a resolution of 1.9 A. The protein is a trimer in a symmetrical cloverleaf arrangement. Each subunit exhibits a new all-helical protein fold in which the two catalytic sulfhydryl groups, Cys-130 and Cys-133, are located near a central cavity in the trimer. The structure supports a mechanism for the alkylhydroperoxidase activity in which Cys-133 is deprotonated by a distant glutamic acid via the relay action of His-137 and a water molecule. The cysteine then reacts with the peroxide to give a sulfenic acid that subsequently forms a disulfide bond with Cys-130. The crystal structure of AhpD identifies a new protein fold relevant to members of this protein family in other organisms. The structural details constitute a potential platform for the design of inhibitors of potential utility as antitubercular agents and suggest that AhpD may have disulfide exchange properties of importance in other areas of M. tuberculosis biology. PMID- 11914370 TI - Structural and conformational analysis of the oxidase to dehydrogenase conversion of xanthine oxidoreductase. AB - Xanthine oxidoreductase (XOR) is a 300-kDa homodimer that can exist as an NAD+ dependent dehydrogenase (XD) or as an O2-dependent oxidase (XO) depending on the oxidation state of its cysteine thiols. Both XD and XO undergo limited cleavage by chymotrypsin and trypsin. Trypsin selectively cleaved both enzyme forms at Lys184, while chymotrypsin cleaved XD primarily at Met181 but cleaved XO at Met181 and at Phe560. Chymotrypsin, but not trypsin, cleavage also prevented the reductive conversion of XO to XD; thus the region surrounding Phe560 appears to be important in the interconversion of the two forms. Size exclusion chromatography showed that disulfide bond formation reduced the hydrodynamic volume of the enzyme, and two-dimensional gel electrophoresis of chymotrypsin digested XO showed significant, disulfide bond-mediated, conformational heterogeneity in the N-terminal third of the enzyme but no evidence of disulfide bonds between the N-terminal and C-terminal regions or between XOR subunits. These results indicate that intrasubunit disulfide bond formation leads to a global conformational change in XOR that results in the exposure of the region surrounding Phe560. Conformational changes within this region in turn appear to play a critical role in the interconversion between the XD and XO forms of the enzyme. PMID- 11914372 TI - Induction of neurite extension and survival in pheochromocytoma cells by the Rit GTPase. AB - The Rit, Rin, and Ric proteins comprise a distinct and evolutionarily conserved subfamily of the Ras-like small G-proteins. Although these proteins share the majority of core effector domain residues with Ras, recent studies suggest that Rit uses novel effector pathways to regulate NIH3T3 cell proliferation and transformation, while the functions of Rin and Ric remain largely unknown. Since we demonstrate that Rit is expressed in neurons, we investigated the role of Rit signaling in promoting the differentiation and survival of pheochromocytoma cells. In this study, we show that expression of constitutively active Rit (RitL79) in PC6 cells results in neuronal differentiation, characterized by the elaboration of an extensive network of neurite-like processes that are morphologically distinct from those mediated by the expression of oncogenic Ras. Although activated Rit fails to stimulate mitogen-activated protein kinase/extracellular-signal-regulated kinase (MAPK/ERK) signaling pathways in COS cells, RitL79 induced the phosphorylation of ERK1/2 in PC6 cells. We also find that Rit-mediated effects on neurite outgrowth can be blocked by co-expression of dominant-negative mutants of C-Raf1 or mitogen-activated protein kinase kinase 1 (MEK1). Moreover, expression of dominant-negative Rit is sufficient to inhibit NGF-induced neurite outgrowth. Expression of active Rit inhibits growth factor withdrawal mediated apoptosis of PC6 cells, but does not induce phosphorylation of Akt/protein kinase B, suggesting that survival does not utilize the phosphatidylinositol 3-kinase (PI3K)/Akt pathway. Instead, pharmacological inhibitors of MEK block Rit-stimulated cell survival. Taken together, these studies suggest that Rit represents a distinct regulatory protein, capable of mediating differentiation and cell survival in PC6 cells using a MEK-dependent signaling pathway to achieve its effects. PMID- 11914373 TI - Assembly of photosystem I. I. Inactivation of the rubA gene encoding a membrane associated rubredoxin in the cyanobacterium Synechococcus sp. PCC 7002 causes a loss of photosystem I activity. AB - A 4.4-kb HindIII fragment, encoding an unusual rubredoxin (denoted RubA), a homolog of the Synechocystis sp. PCC 6803 gene slr2034 and Arabidopsis thaliana HCF136, and the psbEFLJ operon, was cloned from the cyanobacterium Synechococcus sp. PCC 7002. Inactivation of the slr2034 homolog produced a mutant with no detectable phenotype and wild-type photosystem (PS) II levels. Inactivation of the rubA gene of Synechococcus sp. PCC 7002 produced a mutant unable to grow photoautotrophically. RubA and PS I electron transport activity were completely absent in the mutant, although PS II activity was approximately 80% of the wild type level. RubA contains a domain of approximately 50 amino acids with very high similarity to the rubredoxins of anaerobic bacteria and archaea, but it also contains a region of about 50 amino acids that is predicted to form a flexible hinge and a transmembrane alpha-helix at its C terminus. Overproduction of the water-soluble rubredoxin domain in Escherichia coli led to a product with the absorption and EPR spectra of typical rubredoxins. RubA was present in thylakoid but not plasma membranes of cyanobacteria and in chloroplast thylakoids isolated from spinach and Chlamydomonas reinhardtii. Fractionation studies suggest that RubA might transiently associate with PS I monomers, but no evidence for an association with PS I trimers or PS II was observed. PS I levels were significantly lower than in the wild type ( approximately 40%), but trimeric PS I complexes could be isolated from the rubA mutant. These PS I complexes completely lacked the stromal subunits PsaC, PsaD, and PsaE but contained all membrane intrinsic subunits. The three missing proteins could be detected immunologically in whole cells, but their levels were greatly reduced, and degradation products were also detected. Our results indicate that RubA plays a specific role in the biogenesis of PS I. PMID- 11914374 TI - Assembly of photosystem I. II. Rubredoxin is required for the in vivo assembly of F(X) in Synechococcus sp. PCC 7002 as shown by optical and EPR spectroscopy. AB - The rubA gene was insertionally inactivated in Synechococcus sp. PCC 7002, and the properties of photosystem I complexes were characterized spectroscopically. X band EPR spectroscopy at low temperature shows that the three terminal iron sulfur clusters, F(X), F(A), and F(B), are missing in whole cells, thylakoids, and photosystem (PS) I complexes of the rubA mutant. The flash-induced decay kinetics of both P700(+) in the visible and A(1)- in the near-UV show that charge recombination occurs between P700(+) and A(1)- in both thylakoids and PS I complexes. The spin-polarized EPR signal at room temperature from PS I complexes also indicates that forward electron transfer does not occur beyond A(1). In agreement, the spin-polarized X-band EPR spectrum of P700(+) A(1)- at low temperature shows that an electron cycle between A(1)- and P700(+) occurs in a much larger fraction of PS I complexes than in the wild-type, wherein a relatively large fraction of the electrons promoted are irreversibly transferred to [F(A)/F(B)]. The electron spin polarization pattern shows that the orientation of phylloquinone in the PS I complexes is identical to that of the wild type, and out-of-phase, spin-echo modulation spectroscopy shows the same P700(+) to A(1)- center-to-center distance in photosystem I complexes of wild type and the rubA mutant. In contrast to the loss of F(X), F(B), and F(A), the Rieske iron-sulfur protein and the non-heme iron in photosystem II are intact. It is proposed that rubredoxin is specifically required for the assembly of the F(X) iron-sulfur cluster but that F(X) is not required for the biosynthesis of trimeric P700-A(1) cores. Since the PsaC protein requires the presence of F(X) for binding, the absence of F(A) and F(B) may be an indirect result of the absence of F(X). PMID- 11914375 TI - Lactotetraosylceramide, a novel glycosphingolipid receptor for Helicobacter pylori, present in human gastric epithelium. AB - The binding of Helicobacter pylori to glycosphingolipids was examined by binding of (35)S-labeled bacteria to glycosphingolipids on thin-layer chromatograms. In addition to previously reported binding specificities, a selective binding to a non-acid tetraglycosylceramide of human meconium was found. This H. pylori binding glycosphingolipid was isolated and, on the basis of mass spectrometry, proton NMR spectroscopy, and degradation studies, were identified as Galbeta3GlcNAcbeta3Galbeta4Glcbeta1Cer (lactotetraosylceramide). When using non acid glycosphingolipid preparations from human gastric epithelial cells, an identical binding of H. pylori to the tetraglycosylceramide interval was obtained in one of seven samples. Evidence for the presence of lactotetraosylceramide in the binding-active interval was obtained by proton NMR spectroscopy of intact glycosphingolipids and by gas chromatography-electron ionization mass spectrometry of permethylated tetrasaccharides obtained by ceramide glycanase hydrolysis. The lactotetraosylceramide binding property was detected in 65 of 74 H. pylori isolates (88%). Binding of H. pylori to lactotetraosylceramide on thin layer chromatograms was inhibited by preincubation with lactotetraose but not with lactose. Removal of the terminal galactose of lactotetraosylceramide by galactosidase hydrolysis abolished the binding as did hydrazinolysis of the acetamido group of the N-acetylglucosamine. Therefore, Galbeta3GlcNAc is an essential part of the binding epitope. PMID- 11914376 TI - Absence of the I-10 protein segment mediates restricted dimerization of the cartilage-specific fibronectin isoform. AB - The cartilage-specific (V + C)(-) fibronectin isoform does not efficiently heterodimerize with other V-region splice variants of fibronectin. To understand better the structural elements that determine this restricted dimerization profile, a series of truncated fibronectin expression constructs with various internal deletions in the V, III-15, or I-10 segments were constructed and co transfected into COS-7 cells with either the V(+)C(+) or the (V + C)(-) isoform. SDS-PAGE and immunoblot analyses of the resulting conditioned media suggest that the I-10 segment must either be present in both monomeric subunits of fibronectin or absent from both subunits for efficient dimerization to occur. Further studies suggest that the I-10 segment specifically, not simply a balanced number of type I repeats at the carboxyl terminus of each monomeric subunit, plays an important role in determining different fibronectin dimerization patterns. Neither I-11 nor I-12 could be substituted for segment I-10 without significantly reducing the formation of heterodimers. Therefore, absence of segment I-10 explains why (V + C)(-) fibronectin is not found in heterodimeric configurations with other native V-region splice variants in cartilage. The unique dimerization pattern of (V + C)(-) fibronectin does not prevent matrix formation yet is consistent with this isoform having specialized properties in situ that are important for either the structural organization and biomechanical properties of cartilage or the regulation of a chondrocytic phenotype. PMID- 11914377 TI - Role of the G protein gamma subunit in beta gamma complex modulation of phospholipase Cbeta function. AB - The G protein betagamma complex regulates a wide range of effectors, including the phospholipase C isozymes (PLCbetas). Different domains on the beta subunit are known to contact phospholipase Cbeta and affect its regulation. In contrast, the role of the gamma subunit in Gbetagamma modulation of PLCbeta function is not known. Results here show that the gamma subunit C-terminal domain is involved in mediating Gbetagamma interactions with phospholipase Cbeta. Mutations were introduced to alter the position of the post-translational prenyl modification at the C terminus of the gamma subunit with reference to the beta subunit. These mutants were appropriately post-translationally modified with the geranylgeranyl moiety. A deletion that shortened the C-terminal domain, insertions that extended this domain, and a point mutation, F59A, that disrupted the interaction of this domain with the beta subunit were all affected in their ability to activate PLCbeta to varying degrees. All mutants, however, interacted equally effectively with the G(o)alpha subunit. The results indicate that the G protein gamma subunit plays a direct role in the modulation of effector function by the betagamma complex. PMID- 11914378 TI - Regulation of an activated S6 kinase 1 variant reveals a novel mammalian target of rapamycin phosphorylation site. AB - A critical step in S6 kinase 1 (S6K1) activation is Thr(229) phosphorylation in the activation loop by the phosphoinositide-dependent protein kinase (PDK1). Thr(229) phosphorylation requires prior phosphorylation of the Ser/Thr-Pro sites in the autoinhibitory domain and Thr(389) in the linker domain, consistent with PDK1 more effectively catalyzing Thr(229) phosphorylation in a variant harboring acidic residues in these positions (S6K1-E389D(3)E). S6K1-E389D(3)E has high basal activity and exhibits partial resistance to rapamycin and wortmannin, and its activity can be further augmented by mitogens, effects presumably mediated by Thr(229) phosphorylation. However, PDK1-induced Thr(229) phosphorylation is reported to be constitutive rather than phosphatidylinositide 3,4,5-trisphosphate dependent, suggesting that S6K1-E389D(3)E activity is mediated through a distinct site. Here we use phosphospecific antibodies to show that Thr(229) is fully phosphorylated in S6K1-E389D(3)E in the absence of mitogens and that regulation of S6K1-E389D(3)E activity by mitogens, rapamycin, or wortmannin parallels Ser(371) phosphorylation. Consistent with this observation, a dominant interfering allele of the mammalian target of rapamycin, mTOR, inhibits mitogen induced Ser(371) phosphorylation and activation of S6K1-E389D(3)E, whereas wild type mTOR stimulates both responses. Moreover, in vitro mTOR directly phosphorylates Ser(371), and this event modulates Thr(389) phosphorylation by mTOR, compatible with earlier in vivo findings. PMID- 11914379 TI - Novel CD8+ T cell antagonists based on beta 2-microglobulin. AB - The CD8 coreceptor of cytotoxic T lymphocytes binds to a conserved region of major histocompatibility complex class I molecules during recognition of peptide major histocompatibility complex (MHC) class I antigens on the surface of target cells. This event is central to the activation of cytotoxic T lymphocyte (CTL) effector functions. The contribution of the MHC complex class I light chain, beta(2)-microglobulin, to CD8alphaalpha binding is relatively small and is mediated mainly through the lysine residue at position 58. Despite this, using molecular modeling, we predict that its mutation should have a dramatic effect on CD8alphaalpha binding. The predictions are confirmed using surface plasmon resonance binding studies and human CTL activation assays. Surprisingly, the charge-reversing mutation, Lys(58) --> Glu, enhances beta(2)m-MHC class I heavy chain interactions. This mutation also significantly reduces CD8alphaalpha binding and is a potent antagonist of CTL activation. These results suggest a novel approach to CTL-specific therapeutic immunosuppression. PMID- 11914381 TI - Oocyte apoptosis during the transition from ovary-like tissue to testes during sex differentiation of juvenile zebrafish. AB - Large numbers of apoptotic early diplotene oocytes were observed during the transition from ovary-like undifferentiated gonadal tissue to testes during sex differentiation in presumptive males of the zebrafish (Danio rerio). The percentage of terminal-deoxynucleotidyl-transferase-mediated dUTP nick-end labelling (TUNEL)-positive apoptotic oocytes in the gonads of presumptive males was approximately eight- to 12-fold higher than in genetic all-females. By 29 days post-hatching, all oocytes had disappeared from the gonads of presumptive males. In these males, we also observed apoptotic somatic cells in the ovarian cavity between 23 and 35 days post-hatching. Therefore, the disappearance of oocytes and the decomposition of the ovarian cavity caused by apoptosis during sex differentiation were male-specific events. In genetic all-females, apoptosis in a proportion of early diplotene oocytes was found in the undifferentiated gonads at 15-19 days post-hatching, probably as a result of programmed oocyte loss during ovarian development. These findings suggest that oocyte apoptosis is the mechanism of testicular and ovarian differentiation in zebrafish. PMID- 11914382 TI - A comparative study of odorant binding protein genes: differential expression of the PBP1-GOBP2 gene cluster in Manduca sexta (Lepidoptera) and the organization of OBP genes in Drosophila melanogaster (Diptera). AB - Insects discriminate odors using sensory organs called olfactory sensilla, which display a wide range of phenotypes. Sensilla express ensembles of proteins, including odorant binding proteins (OBPs), olfactory receptors (ORs) and odor degrading enzymes (ODEs); odors are thought to be transported to ORs by OBPs and subsequently degraded by ODEs. These proteins belong to multigene families. The unique combinatorial expression of specific members of each of these gene families determines, in part, the phenotype of a sensillum and what odors it can detect. Furthermore, OBPs, ORs and ODEs are expressed in different cell types, suggesting the need for cell-cell communication to coordinate their expression. This report examines the OBP gene family. In Manduca sexta, the genes encoding PBP1Msex and GOBP2Msex are sequenced, shown to be adjacent to one another, and characterized together with OBP gene structures of other lepidoptera and Drosophila melanogaster. Expression of PBP1Msex, GOBP1Msex and GOBP2Msex is characterized in adult male and female antenna and in larval antenna and maxilla. The genomic organization of 25 D. melanogaster OBPs are characterized with respect to gene locus, gene cluster, amino acid sequence similarity, exon conservation and proximity to OR loci, and their sequences are compared with 14 M. sexta OBPs. Sensilla serve as portals of important behavioral information, and genes supporting sensilla function are presumably under significant evolutionary selective pressures. This study provides a basis for studying the evolution of the OBP gene family, the regulatory mechanisms governing the coordinated expression of OBPs, ORs and ODEs, and the processes that determine specific sensillum phenotypes. PMID- 11914384 TI - Cardiorespiratory responses of the common carp (Cyprinus carpio) to severe hypoxia at three acclimation temperatures. AB - In vivo measurements of the cardiovascular responses of anoxia-tolerant teleosts to severe prolonged hypoxia are limited. Here, we report the first direct measurements of cardiac output (Q), heart rate (f(H)) and stroke volume during prolonged severe hypoxia (<0.3 mg O(2) l(-1)) in common carp (Cyprinus carpio L.) that had been acclimated to 6, 10 and 15 degrees C. While routine Q and f(H) values varied with temperature under normoxic conditions (Q(10) values of 1.7 and 2.6, respectively), severe hypoxic exposure significantly depressed f(H) and Q to similar minimum values that were largely independent of acclimation temperature (Q(10) values of 1.2). In contrast, the duration of cardiac depression and the subsequent time period during which carp could tolerate severe hypoxia were inversely related to acclimation temperature (24 h at 6 degrees C, 6 h at 10 degrees C, and 2.5 h at 6 degrees C). Likewise, respiration rate during hypoxia showed a temperature dependence. An unusual finding was that cardiorespiratory status partially recovered during the latter stages of severe hypoxic exposure. We conclude that the cardiorespiratory responses to severe prolonged hypoxia in common carp involved a mixture of temperature-independent, temperature-dependent and time domain phases. PMID- 11914383 TI - NaCl and fluid secretion by the intestine of the teleost Fundulus heteroclitus: involvement of CFTR. AB - Sections of posterior intestine of the euryhaline killifish Fundulus heteroclitus adapted to sea water were stimulated by the calcium ionophore ionomycin (1 micromol l(-1)) in combination with agents to elevate intracellular cyclic AMP levels, 0.5 mmol l(-1) dibutyryl-cyclic AMP (db-cAMP) with 0.1 mmol l(-1) 3 isobutyl-1-methylxanthine (IBMX). Intestinal bag preparations from recently fed animals (but not from overnight unfed animals) changed from fluid absorption (+18.9+/-8.30 microl cm(-2) h(-1), N=8) in the untreated control period to net fluid secretion after stimulation (-7.43+/-1.30 microl cm(-2) h(-1), N=8, P<0.01; means +/- S.E.M.), indicative of the capacity of teleost intestine to undergo secretion. Posterior intestinal pieces mounted in vitro in Ussing-style membrane chambers showed net Cl(-) uptake (+2.245+/-0.633 microequiv cm(-2) h(-1), N=7) that turned to net secretion following stimulation by ionomycin + db-cAMP + IBMX (-3.809+/-1.22 microequiv cm(-2) h(-1), N=7, P<0.01). Mucosal application of the anion channel blocker 1 mmol l(-1) diphenylamine-2-carboxylate (DPC) after ionomycin + db-cAMP + IBMX treatment significantly reduced serosal-to-mucosal unidirectional Cl(-) flux (P<0.001), net Cl(-) flux (P<0.05), short-circuit current (I(sc), P<0.001) and tissue conductance (G(t), P<0.001), while 0.1 mmol l(-1) 4,4'-diisothiocyano-2,2'-stilbene-disulphonic acid (DIDS, a blocker of anion exchange) was without effect. Stimulation by db-cAMP + IBMX (no ionomycin) significantly increased unidirectional fluxes, I(sc) and G(t) but did not produce net Cl(-) secretion. Ionomycin alone produced a transient increase in I(sc) but had no effect on G(t) and caused no significant changes in unidirectional or net Cl(-) fluxes. Addition of db-cAMP + IBMX after ionomycin treatment produced net secretion of Cl(-) and large increases in unidirectional fluxes and G(t). Cystic fibrosis transmembrane conductance regulator (CFTR) was immunocytochemically localized with a monoclonal mouse antibody to the carboxy terminus and found to be present in the cytoplasm and basolateral membranes of all enterocytes and in the brush-border membrane of some cells, whereas NKCC immunofluorescence, demonstrating the presence of the Na(+)/K(+)/2Cl(-) cotransporter, was present in the cytoplasm and brush-border membrane. We conclude that the teleost intestine is capable of salt and fluid secretion only if intracellular Ca(2+) and cyclic AMP pathways are stimulated together and that this secretion appears to involve activation of CFTR ion channels in the apical membrane of a subpopulation of enterocytes. PMID- 11914385 TI - Peripheral oxygen transport in skeletal muscle of Antarctic and sub-Antarctic notothenioid fish. AB - Transcellular oxygen flux was modelled mathematically in the aerobic skeletal muscles of perciform fish species living at widely different temperatures (Antarctica, sub-Antarctica and the Mediterranean Sea). Using structural data derived from stereological analysis of electron micrographs, mean fibre P(O(2)) was calculated on the basis of temperature-corrected rates of mitochondrial respiration and oxygen diffusion. The mean muscle fibre diameter (MFD) among Antarctic notothenioids was in the range 17-61 microm and mitochondrial volume density, Vv(mit,f), was 0.27-0.53, but capillary-to-fibre ratio varied only between 1.2 and 1.5. For a mean capillary P(O(2)) of 6 kPa, the model predicted a mean tissue P(O(2)) in the range 0.7-5.8 kPa at the estimated maximum aerobic capacity (M(O(2)max)). The lowest levels of tissue oxygenation were found in the pectoral muscle fibres of the icefish Chaenocephalus aceratus, which lacks the respiratory pigments haemoglobin and myoglobin. Red-blooded notothenioids found in the sub-Antarctic had a similar muscle fine structure to those caught south of the Antarctic Convergence, with an MFD of 20-41 microm and Vv(mit,f) of 0.27 0.33, resulting in an estimated mean P(O(2)) of 4-5 kPa at M(O(2)max). Mean tissue P(O(2)) in the sub-Antarctic icefish Champsocephalus esox, with greater MFD and Vv(mit,f), 56 microm and 0.51, respectively, was calculated to exceed 1 kPa at winter temperatures (4 degrees C), although oxidative metabolism was predicted to be impaired at the summer maximum of 10 degrees C. At the high end of the thermal range, related perciform species from the Mediterranean had a negligible drop in intracellular P(O(2)) across their small-diameter fibres, to a minimum of 5.4 kPa, comparable with that predicted for Trematomus newnesi from the Antarctic (5.6 kPa) with a similar MFD. These data suggest that, within a single phylogenetic group, integrative structural adaptations potentially enable a similar degree of tissue oxygenation over a 20 degrees C range of environmental temperature in the red-blooded notothenioids, and that this is compromised by the lack of respiratory pigments in the icefishes. The mean capillary radius was 1.5 times greater in the two icefish than in the other notothenioids, and the model simulations indicate that the evolution of wide-bore capillaries is essential to maintain tissue oxygenation in the absence of respiratory pigments. PMID- 11914386 TI - Lipophorin-facilitated hydrocarbon uptake by oocytes in the German cockroach Blattella germanica (L.). AB - Large amounts of hydrocarbons accumulate during vitellogenesis in the developing basal oocytes of the German cockroach Blattella germanica (L.), and all ovarian hydrocarbons are deposited into an egg case (ootheca) during oviposition. Hydrocarbons are not synthesized by the ovaries, but are delivered by hemolymph lipoproteins and accumulate within the basal oocytes. A native B. germanica hydrocarbon, [(3)H]3,11-dimethylnonacosane, injected into adult females of various ages, was taken up by the ovaries in relation to oocyte growth. Ovarian uptake of the hydrocarbon was low in day 0-1 females, increased dramatically between days 3 and 6 and declined sharply through oviposition on day 8-9; ovarian uptake of the hydrocarbon was low during a 21-day pregnancy that followed. [1 (14)C]Propionate, which becomes incorporated into methyl-branched hydrocarbons, was injected into 5-day-old vitellogenic females to monitor the de novo biosynthesis of hydrocarbons and the time course of hydrocarbon deposition in the ovary. Propionate was rapidly incorporated into hydrocarbons within 4 h. Hydrocarbon uptake by the ovaries, however, was three times higher 24 h after injection than 4 h after injection, showing that hydrocarbons are slowly and continuously deposited in oocytes. This result was confirmed with topical application of [(3)H]3,11-dimethylnonacosane: ovarian uptake was three times higher after 24 h than after 4 h. In vitro incubations of sternites, which synthesize hydrocarbons, with [(14)C]propionate and ovaries, showed that both hemolymph and purified high-density lipophorin facilitated ovarian uptake of newly synthesized hydrocarbons; maximum uptake occurred with 10 % hemolymph or 1 mg ml(-1) high-density lipophorin. These results were confirmed with sternites treated with [(3)H]3,11-dimethylnonacosane and co-incubated with ovaries. This is the first report to show that copious amounts of maternal hydrocarbons are provisioned in oocytes and to demonstrate the existence of a hydrocarbon transport pathway involving hemolymph high-density lipophorin. PMID- 11914387 TI - Metabolic rate and respiratory gas-exchange patterns in tenebrionid beetles from the Negev Highlands, Israel. AB - This study correlates the pattern of external gas exchange with the diel activity of nine species of tenebrionid beetle from the Negev Desert, Israel. The study species are active throughout the summer months when daytime temperatures are high and no rain falls. There were no differences in standard metabolic rate, determined by flow-through respirometry, among the nine species. All the nocturnally active beetles exhibited a form of continuous respiration, whereas the two diurnally active and one crepuscular species exhibited a cyclic form of respiration referred to as the discontinuous gas-exchange cycle (DGC). The DGCs recorded have a long flutter period consisting of miniature ventilations, and 29 48 % of the total CO(2) output occurred during this period. In this study, the flutter period played an important role in the modulation of metabolic rate, in contrast to other studies in which the burst period has been shown to be important. We suggest that the long flutter period is important in reducing respiratory water loss in arid-dwelling arthropods. This study lends support to the hypothesis that discontinuous gas exchange is important in reducing respiratory water loss from beetles that need to minimise dessication because of the high water vapour pressure gradient they experience. If the use of underground burrows were responsible for the evolution of discontinuous gas exchange, then we would expect all nine tenebrionid species to use DGCs since both the nocturnally and diurnally active species bury in the sand during periods of inactivity. We conclude that the activity patterns of the beetles are more important than their habitat associations in designating the type of respiration used. PMID- 11914388 TI - Luminescence in ophiuroids (Echinodermata) does not share a common nervous control in all species. AB - Study of the control mechanisms of light emission in invertebrates shows the involvement of several neurotransmitters. In ophiuroids, only one species (Amphipholis squamata) has so far been characterized for luminescence control, which seems to be cholinergic, with an influence of several excitatory and inhibitory neuromodulators (amino acids, catecholamines, neuropeptides S1 and S2, purines). The aim of this work is to investigate the nature of control mechanisms of light emission in three luminous ophiuroid species, A. filiformis, O. aranea and O. californica, in order to see whether or not they share common mechanisms. Luminescence induced by general depolarisation of tissues using KCl (200 mmol l( 1)) shows different patterns, according to species. Only A. filiformis emits light in response to acetylcholine. In this species, the involvement of both muscarinic and nicotinic receptors is proposed, since atropine and tubocurarine (at 10(-3) mol l(-1)) inhibited 99 % and 71 %, respectively, of the light emitted. Study of the subtypes of cholinergic receptors involved in photogenesis revealed that several subtypes of muscarinic receptors might be involved. It was also clearly shown that ophiuroids did not share a common mechanism of nervous control of luminescence in all species. PMID- 11914389 TI - The use of landmarks and panoramic context in the performance of local vectors by navigating honeybees. AB - Bees seem to use landmarks to segment familiar routes. They can associate, with a landmark, a memory that encodes the direction and distance of the path segment between that landmark and the next. The expression of the memory results in the performance of a local vector matching the distance and direction of the path segment. The memories of path segments appear to be 'chained' together, so that the performance of one local vector is sometimes sufficient to elicit the subsequent local vector, even in the absence of the associated landmark. We have investigated the effect of visual panoramic context on the expression of local vectors. Bees were trained to fly along a narrow channel to collect sucrose from a feeder positioned partway along it. Panoramic context was provided by various types of patterning on the walls. The channel was partitioned into different segments using landmarks of two kinds: a boundary landmark that marked a change in the pattern on one or both side-walls of the channel, and an isolated landmark, consisting of a baffle through which the bee passed, for which the wall pattern was the same before as after. In tests, we removed the feeder and analysed the search distribution of the bees for various arrangements of landmarks. Altering the spatial relationship between landmarks has different consequences for the two types of landmark. If the final boundary landmark is shifted, the centre of the search distribution shifts by approximately the same amount. Changes in the position of an isolated landmark have a weaker effect. In the absence of the final context, the search is disrupted. We suggest that for local vectors to be expressed the surrounding panoramic context needs to be appropriate. A comparison of search patterns from two different training configurations of landmarks supports the hypothesis that local vector memories merely encode route segments and that global positional coordinates are not linked to landmark memories. PMID- 11914390 TI - Anoxia induces thermotolerance in the locust flight system. AB - Heat shock and anoxia are environmental stresses that are known to trigger similar cellular responses. In this study, we used the locust to examine stress cross-tolerance by investigating the consequences of a prior anoxic stress on the effects of a subsequent high-temperature stress. Anoxic stress and heat shock induced thermotolerance by increasing the ability of intact locusts to survive normally lethal temperatures. To determine whether induced thermotolerance observed in the intact animal was correlated with electrophysiological changes, we measured whole-cell K(+) currents and action potentials from locust neurons. K(+) currents recorded from thoracic neuron somata were reduced after anoxic stress and decreased with increases in temperature. Prior anoxic stress and heat shock increased the upper temperature limit for generation of an action potential during a subsequent heat stress. Although anoxia induced thermotolerance in the locust flight system, a prior heat shock did not protect locusts from a subsequent anoxic stress. To determine whether changes in bioenergetic status were implicated in whole-animal cross-tolerance, phosphagen levels and rates of mitochondrial respiration were assayed. Heat shock alone had no effect on bioenergetic status. Prior heat shock allowed rapid recovery after normally lethal heat stress but afforded no protection after a subsequent anoxic stress. Heat shock also afforded no protection against disruption of bioenergetic status after a subsequent exercise stress. These metabolite studies are consistent with the electrophysiological data that demonstrate that a prior exposure to anoxia can have protective effects against high-temperature stress but that heat shock does not induce tolerance to anoxia. PMID- 11914391 TI - Differing mechanisms of cold-induced changes in capillary supply in m. tibialis anterior of rats and hamsters. AB - The physiological, metabolic and anatomical adaptations of skeletal muscle to chronic cold exposure were investigated in Wistar rats (Rattus norvegicus), a species that defends core temperature, and Syrian hamsters (Mesocricetus auratus), which may adopt a lower set point under unfavourable conditions. Animals were exposed to a simulated onset of winter in an environmental chamber, progressively shortening photoperiod and reducing temperature from 12 h:12 h L:D and 22 degrees C to 1 h:23 h L:D and 5 degrees C over 4 weeks. The animals were left at 4 degrees C for a further 4 weeks to complete the process of cold acclimation. M. tibialis anterior from control (euthermic) and cold-acclimated animals of similar mass showed a significant hyperactivity-induced hypertrophy in the rat, but a small disuse atrophy in the hamster. Little evidence was found for interconversion among fibre types in skeletal muscle on cold-acclimation, and only modest differences were seen in activity of oxidative or glycolytic enzymes in either species. However, adjustments in Type II fibre size paralleled the muscle hypertrophy in rat and atrophy in hamster. Cold-induced angiogenesis was present in the rat, averaging a 28 % increase in capillary-to-fibre ratio (C:F) but, as this was balanced by fibre hypertrophy across the whole muscle, there was no change in capillary density (CD). In contrast, the C:F was similar in both groups of hamsters, whereas CD rose by 33 % in line with fibre atrophy. Within distinct regions of the m. tibialis anterior, there was a correlation between angiogenesis and fibre size in rats, in which oxygen diffusion distance increased, but not in hamsters, in which there was a reduced oxygen diffusion distance. Consequently, the change in C:F was greatest (39 %) in the glycolytic cortex region of the m. tibialis anterior in rats. We conclude that non hibernator and hibernator rodents improve peripheral oxygen transport following cold-acclimation by different mechanisms. In rats, an increase in fibre girth was accompanied by a true angiogenesis, while the improved apparent capillary supply in hamsters was due to smaller fibre diameters. These responses are consistent with the strategies of resisting and accommodating, respectively, an annual fall in environmental temperature. PMID- 11914392 TI - Effects of metamorphosis on the aquatic escape response of the two-lined salamander (Eurycea bislineata). AB - Although numerous studies have described the escape kinematics of fishes, little is known about the aquatic escape responses of salamanders. We compare the escape kinematics of larval and adult Eurycea bislineata, the two-lined salamander, to examine the effects of metamorphosis on aquatic escape performance. We hypothesize that shape changes associated with resorption of the larval tail fin at metamorphosis will affect aquatic locomotor performance. Escape responses were recorded using high-speed video, and the effects of life stage and total length on escape kinematics were analyzed statistically using analysis of covariance. Our results show that both larval and adult E. bislineata use a two-stage escape response (similar to the C-starts of fishes) that consists of a preparatory (stage 1) and a propulsive (stage 2) stroke. The duration of both kinematic stages and the distance traveled during stage 2 increased with total length. Both larval and adult E. bislineata had final escape trajectories that were directed away from the stimulus. The main kinematic difference between larvae and adults is that adults exhibit significantly greater maximum curvature during stage 1. Total escape duration and the distance traveled during stage 2 did not differ significantly between larvae and adults. Despite the significantly lower tail aspect ratio of adults, we found no significant decrease in the overall escape performance of adult E. bislineata. Our results suggest that adults may compensate for the decrease in tail aspect ratio by increasing their maximum curvature. These findings do not support the hypothesis that larvae exhibit better locomotor performance than adults as a result of stronger selective pressures on early life stages. PMID- 11914393 TI - Dual antennular chemosensory pathways mediate odor-associative learning and odor discrimination in the Caribbean spiny lobster Panulirus argus. AB - Chemosensory neurons in the antennular flagella of lobsters mediate long-range responses to chemicals. These neurons are part of two parallel chemosensory pathways with different peripheral and central components. Aesthetasc sensilla on the lateral flagella are innervated by chemosensory neurons that project to the olfactory lobes. A diversity of other 'non-aesthetasc' sensilla on both lateral and medial flagella are innervated by mechano- and chemosensory neurons, and most of these non-aesthetasc neurons project to the lateral antennular neuropils. We investigated the roles of these two pathways in odor-associative learning and odor discrimination by selectively removing either aesthetasc or non-aesthetasc sensilla from the spiny lobster Panulirus argus. Lobsters lacking both aesthetasc and non-aesthetasc antennular sensilla show very reduced or no odor-mediated searching behavior. We associatively conditioned lobsters using two paradigms: aversive conditioning with generalization testing (which reveals the similarity in the lobsters' perception of odorants) and discrimination conditioning (which reveals the lobsters' ability to discriminate odorants). Sham-control intact lobsters performed these tasks well, as did lobsters lacking either aesthetascs or non-aesthetasc setae. There was a strong but statistically non-significant trend that lobsters lacking either aesthetascs or non-aesthetasc setae generalized more between complex odor mixtures than did intact lobsters. After aversive conditioning with generalization testing, aesthetasc-ablated lobsters had more difficulty discriminating among the most closely related complex mixtures than did intact or non-aesthetasc-ablated lobsters. However, after discrimination conditioning, aesthetasc-ablated lobsters were as proficient as intact animals in discriminating highly similar mixtures. These results indicate overlap and redundancy in the function of these two chemosensory pathways in odor associative learning and odor discrimination, but these pathways also complement each other to enable better discrimination. This study presents the first evidence for a role of non-aesthetasc chemosensory neurons in complex odor mediated behaviors such as learning and discrimination. PMID- 11914394 TI - Cn11, the first example of a scorpion toxin that is a true blocker of Na(+) currents in crayfish neurons. AB - A novel crustacean toxin (Cn11) was isolated and characterized from the venom of the Mexican scorpion Centruroides noxius Hoffmann. It contains 63 amino acid residues and is stabilized by four disulphide bridges. It is lethal to crustaceans (Cambarellus montezumae), less toxic to insects (crickets) and non toxic to mammals (mice) at the doses assayed. In neurons isolated from the X organ-sinus gland system of the crayfish Procambarus clarkii, it blocks the Na(+) currents with an estimated K(m) of 320 nmol l(-1), without affecting the Ca(2+) and K(+) currents. The voltage-gated tetrodotoxin-sensitive Na(+) current was recorded from X organ neurons in culture 24 h after plating using the whole-cell clamp configuration. The Na(+) current was isolated by blocking Ca(2+) currents with Cd(2+) and Cs(+) and K(+) currents with tetraethylammonium and 4 aminopyridine. Under control conditions, the Na(+) currents were activated at -40 mV with a maximum amplitude at 0 mV. In the presence of 1 micromol l(-1) Cn11, the Na(+) current amplitude was reduced by 75 % without apparent modifications to the gating mechanism. These findings suggest that Cn11 selectively blocks a Na(+) channel. It is the first representative of a new group of scorpion toxins specific for this molecular target. PMID- 11914395 TI - An important first step: a standard curriculum for the neurology clerkship. PMID- 11914396 TI - ALADIN, but where's the genie? PMID- 11914397 TI - The neurology clerkship core curriculum. AB - Neurologic symptoms are common in all practice settings, and neurologic diseases comprise a large and increasing proportion of health care expenditures and global disease burden. Consequently, the training of all physicians should prepare them to recognize patients who may have neurologic disease, and to take the initial steps in evaluating and managing those patients. We present a core curriculum outlining the clinical neurology skills and knowledge necessary to achieve that degree of preparation. The curriculum emphasizes general principles and a systematic approach to patients with neurologic symptoms and signs. The ability to perform and interpret the neurologic examination is fundamental to that approach, so the curriculum delineates the essential components of the examination in three different clinical settings. The focus of the curriculum is on symptom-based rather than disease-based learning. The only specific diseases selected for inclusion are conditions that are common or require urgent management. This curriculum has been approved by the national organization of neurology clerkship directors and endorsed by the major national professional organizations of neurologists. It is intended as a template for planning a neurology clerkship and as a benchmark for evaluating existing clerkships. It should be especially helpful to clerkship directors, neurology chairs, deans of medical education, and members of external accreditation groups. PMID- 11914398 TI - Cerebral cavernous malformations: mutations in Krit1. AB - OBJECTIVE: To find mutations in the recently identified additional exons of the Krit1 gene that causes CCM1, a disease characterized by the formation of cerebral cavernous malformations (CCM). To determine the relative frequency with which Krit1 mutations cause CCM as well as recharacterize the mutations reported in the literature. METHODS: Twenty-seven families and 11 apparently sporadic individuals affected with CCM were screened for mutations in the Krit1 gene. The gene was screened by single stranded conformation polymorphism, and variants were sequenced. Familial segregation of the mutations was determined. RESULTS: In familial samples, two new mutations in the novel upstream exons and six additional mutations in the previously identified exons were identified. No mutation was found in any of the sporadic individuals. CONCLUSIONS: Results demonstrate that the frequency of mutations found in Krit1 is 47% in the families studied and the frequency may increase as more mutations are detected. Mutations are evenly distributed in the gene and do not seem to be limited to structural domains present in Krit1. This is in accordance with the model that Krit1 could be a tumor suppressor gene. PMID- 11914399 TI - Neuropsychological and psychiatric sequelae of pallidotomy for PD: clinical trial findings. AB - OBJECTIVE: To evaluate the neuropsychological and psychiatric sequelae of unilateral posterior pallidotomy for treatment of PD. METHODS: Patients with idiopathic PD completed baseline and 3- and 6-month assessments after random assignment to an immediate surgery (n = 17) or medical management (n = 16) group. RESULTS: Compared with the medical management group, the immediate surgery group with single lesions centered on the posterior internal pallidum showed superior naming and response inhibition, better verbal recall at 6 months, but greater distractibility, a tendency toward lower phonemic fluency, and a transient (3 months' only) semantic fluency deficit. The group with left lesions had more neuropsychological deficits than the group with right lesions or the medical management group, although these occurred mainly at 3 (but not 6) months. At 6 months, the patients with left lesions showed better verbal memory retention than the patients with right lesions. On most measures, the pattern of individual clinical change did not differ as a function of surgery or lesion laterality, with the exception of a higher frequency of decline in phonemic fluency in the patients with left lesions at 6 months. Although psychiatric status did not change overall, a history of depression tended to increase the risk of a depressive episode following surgery. CONCLUSIONS: Well-targeted, uncomplicated, unilateral pallidotomy does not produce overall neuropsychological or psychiatric change, although there are subtle changes on specific measures sensitive to frontal lobe function. PMID- 11914400 TI - Evidence for widespread movement-associated functional MRI changes in patients with PPMS. AB - BACKGROUND: Previous work has suggested that functional reorganization of cortical motor areas might have a role in limiting the motor deficits in patients with MS. OBJECTIVE: To test whether movement-associated cortical changes in MS might extend beyond the "classic" motor areas and involve sites for multimodal integration. METHODS: fMRI was used to assess patterns of brain activations associated with 3 different motor tasks in 30 right-handed patients with primary progressive MS (PPMS) and variable degrees of motor impairment, which were compared with those from 15 right-handed, sex- and age-matched control subjects. RESULTS: Compared with control subjects, patients with MS showed increased activation of brain regions within both traditional motor planning and execution regions (including the supplementary motor area and the cingulate motor area), the insula (a region implicated in sensory processing), and several multimodal cortical regions in the temporal, parietal, and occipital lobes. In patients, the extent of the fMRI activations was strongly correlated with MR lesion burden (r ranging from 0.70 to 0.86, p < 0.001). CONCLUSIONS: This study shows that movement-associated cortical activation in patients with PPMS is widely distributed and also involves multimodal "nonmotor" cortical networks. It also suggests that adaptive cortical reorganization might be one of the mechanisms limiting the clinical impact of MS in the progressive phases of the disease. PMID- 11914401 TI - Ciguatera fish poisoning: a double-blind randomized trial of mannitol therapy. AB - BACKGROUND: Ciguatera poisoning (CP) is worldwide the most common fish-borne illness and one of the most common forms of nonbacterial food poisoning. IV mannitol is considered the treatment of choice for CP but has not been evaluated in a double-blind randomized trial. METHODS: A prospective clinical study of 50 patients with CP on Rarotonga, Cook Islands, was conducted to better define the neurologic picture of CP and to study the effect of mannitol or normal saline under double-blind randomized conditions over a 24-hour period. RESULTS: The neurologic presentation of CP was that of a predominantly sensory, length dependent polyneuropathy, with preferential small-fiber involvement. Motor paresis, cranial nerve dysfunction, and CNS abnormalities were absent but for a rare mild transitory cerebellar syndrome. At 24 hours, 96% of mannitol-treated patients and 92% of normal saline-treated patients had some improvement of symptoms (p = 1.0), whereas 12% and 24% of patients in each group were asymptomatic (p = 0.46). By 24 hours, the prevalence of the various polyneuropathic symptoms and signs was reduced roughly by half in both groups. Discomfort or pain along the vein used for infusion was more frequent in the mannitol group (84%) than in the normal saline group (36%) (p = 0.0015). CONCLUSION: Mannitol was not superior to normal saline in relieving symptoms and signs of CP at 24 hours in this study population but had more side effects. These results do not support single-dose mannitol as standard treatment for CP. PMID- 11914402 TI - Association of homozygous 7048G7049 variant in the intron six of Nurr1 gene with Parkinson's disease. AB - OBJECTIVE: To determine whether the Nurr1 gene, which is critical for the development and maintenance of nigral dopaminergic neurons, is a risk factor associated with PD. BACKGROUND: The Nurrl gene is highly expressed in the dopaminergic neurons in the midbrain. Knockout of the gene results in agenesis of nigral dopaminergic neurons and heterozygous knockout mice increases 1-methyl-4 phenyl-1,2,3,6-tetrahydropyridine (MPTP)-induced neurotoxicity. METHODS: This study included 105 patients with familial PD (fPD) and 120 patients with sporadic PD (sPD) and 221 age-matched healthy control subjects. The polymorphisms and mutations of the Nurr1 gene in patients with PD were initially examined by heteroduplex analysis and sequencing analysis from PCR-amplified Nurr1 gene fragments. A polymorphism in the BseRI restriction site was identified, and a relatively large-scale analysis then was conducted by three independent investigators who were blinded to the clinical status of the subjects. RESULTS: A homozygous 7048G7049 polymorphism was found in intron 6 of the Nurr1 gene, which was significantly higher in fPD (10/105; 9.5%) and in sPD (5/120; 4.2%) compared with healthy control subjects (2/221; 0.9%). The mean age and the SD at onset of these homozygote patients with PD was 52 +/- 15 years for fPD and 46 +/- 7 years for sPD. The clinical features of these homozygote patients with PD did not differ from those of typical PD. CONCLUSIONS: The homozygote polymorphism of 7048G7049 in intron 6 of the Nurr1 gene is associated with typical PD. PMID- 11914403 TI - Migraine in the United States: epidemiology and patterns of health care use. AB - OBJECTIVE: To determine the prevalence and distribution of migraine in the United States as well as current patterns of health care use. METHODS: A random-digit dial, computer-assisted telephone interview (CATI) survey was conducted in Philadelphia County, PA, in 1998. The CATI identifies individuals with migraine (categories 1.1 and 1.2) as defined by the diagnostic criteria of the International Headache Society with high sensitivity (85%) and specificity (96%). Interviews were completed in 4,376 subjects to identify 568 with migraine. Those with 6 or more attacks per year (n = 410) were invited to participate in a follow up interview about health care utilization and family impact of migraine; 246 (60.0%) participated. RESULTS: The 1-year prevalence of migraine was 17.2% in females and 6.0% in males. Prevalence was highest between the ages of 30 and 49. Whereas 48% of migraine sufferers had seen a doctor for headache within the last year (current consulters), 31% had never done so in their lifetimes and 21% had not seen a doctor for headache for at least 1 year (lapsed consulters). Of current or lapsed consulters, 73% reported a physician-made diagnosis of migraine; treatments varied. Of all migraine sufferers, 49% were treated with over-the-counter medications only, 23% with prescription medication only, 23% with both, and 5% with no medications at all. CONCLUSION: Relative to prior cross sectional surveys, epidemiologic profiles for migraine have remained stable in the United States over the last decade. Self-reported rates of current medical consultation have more than doubled. Moderate increases were seen in the percentage of migraine sufferers who use prescription medications and in the likelihood of receiving a physician diagnosis of migraine. PMID- 11914404 TI - Expression of JC virus T-antigen in a patient with MS and glioblastoma multiforme. AB - OBJECTIVE: To investigate the presence of human polyomavirus JC virus genome and the expression of the viral oncoprotein T-antigen in neoplastic cells of a patient with MS and a glioblastoma multiforme. BACKGROUND: The postmortem examination of an immunocompetent patient with a neurologic disorder revealed the concurrence of MS plaques in the white matter of the brain and a glioblastoma multiforme in the region of the thalamus. METHODS AND RESULTS: PCR analysis of DNA from demyelinated plaques and the tumor area using primers derived from specific regions of the JC virus genome revealed the presence of viral DNA corresponding to the viral early and late genes. Further examination of the samples for the JC virus regulatory region identified the presence of sequences identical to JC virus Mad-4 and JC virus W1 viral isolates in the tumor and the demyelinated regions. Results from immunohistochemistry showed the detection of the viral early protein, T-antigen, and the cellular tumor suppressor protein, p53, in the nuclei of neoplastic cells. Interestingly, expression of T-antigen, but not p53, was observed in neurofilament-positive cells with neuronal morphology and in glial fibrillary acidic protein-positive astrocytes in the cortex juxtaposed to the MS plaques. Examination of viral late gene expression by immunohistochemistry showed no evidence for viral capsid proteins, thus ruling out productive replication of JC virus in the tumor and MS demyelinated plaques. CONCLUSIONS: These observations provide molecular and clinical evidence of the association of JC virus in the brain of a patient with concurrent glioblastoma multiforme and MS. PMID- 11914405 TI - Proton MR spectroscopic imaging predicts lesion progression on MRI in X-linked adrenoleukodystrophy. AB - BACKGROUND: The phenotypic expression of X-linked adrenoleukodystrophy (X-ALD) ranges from the rapidly progressive childhood cerebral form to the milder adrenomyeloneuropathy in adults. It is not possible to predict phenotype by mutation analysis or biochemical assays. Multislice proton MRS imaging (MRSI) has previously detected more extensive brain abnormalities in X-ALD than conventional MRI, which has been suggested to predict impending demyelination. However, the significance of these changes is unclear. OBJECTIVE: The purpose of this study was to determine the long-term sensitivity and specificity of MRSI for disease progression in X-ALD. METHODS: Twenty-five patients with X-ALD were investigated (average age, 15 years; range, 2-43 years) with MRI and proton MRSI at baseline and follow-up MRI over a mean period of 3.5 years. Eight patients had normal MRI findings at baseline and on follow-up (noncerebral group), 11 had abnormal MRI at baseline and no change on follow-up (cerebral nonprogressive group), and 6 had progressive MRI abnormalities (cerebral progressive group). On MRSI, voxels were analyzed in the normal MRI-appearing perilesional white matter, or in the corresponding area in the noncerebral group. RESULTS: The concentration ratio of N-acetylaspartate (NAA) to choline was the most sensitive indicator of disease progression. The average NAA/choline ratio was 5.99 for the noncerebral group, 5.75 for the cerebral nonprogressive group, and 3.74 for the cerebral progressive group (p = 0.002). At a cut-off point of 5.0, the NAA/choline ratio predicted disease progression in all patients with six cerebral progressive disease (sensitivity 100%). The specificity was 83%, the positive predictive value was 66%, and the negative predictive value was 100%. CONCLUSIONS: Multislice proton MRS imaging is able to identify impending or beginning degeneration in white matter that still appears normal on conventional MRI. Multislice proton MRSI may be a suitable technique for the prediction of lesion progression on MRI in X linked adrenoleukodystrophy. PMID- 11914406 TI - Increased familial risk of the psychotic phenotype of Alzheimer disease. AB - BACKGROUND: Psychotic symptoms in patients with AD (AD with psychosis [AD+P]) define a phenotype characterized by more rapid cognitive and functional decline and a liability to aggressive behaviors. OBJECTIVE: To determine if AD+P aggregates within families. METHODS: Case-control study of AD+P frequency in 461 siblings of 371 probands diagnosed with AD. All siblings were ascertained as part of a genetic investigation and also were diagnosed with AD. Statistical analysis used Generalized Estimating Equations to adjust for clustering within families. RESULTS: AD+P in probands was associated with a significantly increased risk for AD+P in family members (OR, 2.41; 95% CI 1.46-4.0; p = 0.0006). The correlation among siblings for AD+P status was modest: 0.16. CONCLUSION: AD+P demonstrates familial aggregation. Further studies are required to investigate a possible genetic basis of AD+P. PMID- 11914407 TI - Prevalence and clinical importance of sleep apnea in the first night after cerebral infarction. AB - OBJECTIVE: To determine the prevalence of sleep apnea (SA) during the first night after hemispheric ischemic stroke and its influence on clinical presentation, course, and functional outcome at 6 months. METHODS: The first night after cerebral infarction onset, 50 patients underwent polysomnography (PSG) followed by oximetry during the next 24 hours. Neurologic severity and early worsening were assessed by the Scandinavian Stroke Scale and outcome by the Barthel Index. Patients were evaluated on admission, on the third day, at discharge, and at 1, 3, and 6 months. RESULTS: There were 30 males and 20 females with a mean age of 66.8 +/- 9.5 years. Latency between stroke onset and PSG was 11.6 +/- 5.3 hours. Thirty-one (62%) subjects had SA (apnea-hypopnea index [AHI] > or = 10). Of these, 23 (46%) had an AHI > or =20 and 21 (42%) an AHI > or =25. Sleep-related stroke onset occurred in 24 (48%) patients and was predicted only by an AHI > or =25 on logistic regression analysis. SA was related to early neurologic worsening and oxyhemoglobin desaturations but not to sleep history before stroke onset, infarct topography and size, neurologic severity, or functional outcome. Early neurologic worsening was found in 15 (30%) patients, and logistic regression analysis identified SA and serum glucose as its independent predictors. CONCLUSIONS: SA is frequent during the first night after cerebral infarction (62%) and is associated with early neurologic worsening but not with functional outcome at 6 months. Cerebral infarction onset during sleep is associated with the presence of moderate to severe SA (AHI > or = 25). PMID- 11914408 TI - Familial periventricular heterotopia: missense and distal truncating mutations of the FLN1 gene. AB - OBJECTIVE: To examine the clinical and MRI associations in bilateral periventricular nodular heterotopia (BPNH) (MIM # 300049) in two families segregating a missense mutation and a C-terminal deletion of the filamin 1(FLN1) gene. BACKGROUND: Classical familial BPNH, an X-linked dominant disorder, has been associated with protein truncations or splicing mutations, which tend to cluster at the N-terminal of the FLN1 protein, causing severe predicted loss of the protein function. The clinical syndrome includes symmetrical contiguous nodular heterotopia lining the lateral ventricles, epilepsy, mild retardation to normal cognitive level in affected females, and prenatal lethality in hemizygous boys. METHODS: Clinical examination, cognitive testing, MRI, mutation analysis (direct sequencing, single-strand conformation polymorphism) in seven patients from two families with BPNH. RESULTS: In Family 1, harboring an A > T change in exon 2 (E82V), heterotopic nodules were few, asymmetric, and noncontiguous. Five boys born from affected females had died unexpectedly early in life. In Family 2, harboring an 8 base pair deletion in exon 47 (7627_7634del TGTGCCCC), heterotopic nodules were thick and contiguous. Affected females in both families showed normal to borderline IQ and epilepsy. CONCLUSION: Missense mutations and distal truncations consistent with partial loss of FLN1 function cause familial BPNH with the classical clinical phenotype including epilepsy and mild mental retardation, if any. However, missense mutations have milder anatomic consequences in affected females and are possibly compatible with live birth but short survival of boys. PMID- 11914409 TI - Early onset autosomal dominant dementia with ataxia, extrapyramidal features, and epilepsy. AB - OBJECTIVE: To perform a clinical and molecular study of a large autosomal dominant family with a complex neurologic syndrome that comprises early-onset dementia, extrapyramidal and cerebellar features, and epilepsy. BACKGROUND: Early onset forms of dementia often are caused by genetic factors. Mutations of three different genes-amyloid precursor protein (APP), presenilin 1 (PS-1), presenilin 2 (PS-2)-have been found in early-onset autosomal dominant forms of AD, of the human microtubule associated-protein tau gene (MAPT) in frontotemporal dementia and parkinsonism linked to chromosome 17 (FTDP-17), of the BRI gene in familial British dementia, of the PI12 gene in familial encephalopathy with neuroserpin inclusion bodies. Linkage to chromosome 3 has been found in familial nonspecific dementia (FND) and linkage to chromosome 20 has been found in Huntington disease (HD)-like neurodegenerative disease. Dementia may be a feature of other neurodegenerative diseases such as HD, dentatorubro-pallidoluysian atrophy (DRPLA), diseases caused by mutations of the prion protein gene (PRNP), spinocerebellar ataxias (SCA), and familial parkinsonism. METHODS: A southern Italian family with autosomal dominant dementia-plus was observed. The family includes 57 individuals in 5 generations (14 affected, 7 personally observed). The authors performed linkage analysis to APP, PS-1, PS-2, FTDP-17, BRI, PI12, FND, HD-like, SCA4, SCA5, SCA10, SCA11, SCA13, PARK1, PARK2, PARK3 loci; direct mutation analysis of HD, DRPLA, SCA1, SCA2, SCA3, SCA6, SCA7, SCA8, SCA12, and PRNP genes; and sequencing of the PRNP open reading frame. RESULTS: Linkage to the examined loci was excluded. All of the direct mutation analyses were negative excluding mutations in the examined genes. CONCLUSIONS: This family has a peculiar phenotype and molecular analyses excluded genes known to cause hereditary dementias. PMID- 11914410 TI - Selective reduction of N-acetylaspartate in medial temporal and parietal lobes in AD. AB - BACKGROUND: Both AD and normal aging cause brain atrophy, limiting the ability of MRI to distinguish between AD and age-related brain tissue loss. MRS imaging (MRSI) measures the neuronal marker N-acetylaspartate (NAA), which could help assess brain change in AD and aging. OBJECTIVES: To determine the effects of AD on concentrations of NAA, and choline- and creatine-containing compounds in different brain regions and to assess the extent NAA in combination with volume measurements by MRI improves discrimination between AD patients and cognitively normal subjects. METHODS: Fifty-six patients with AD (mean age: 75.6 +/- 8.0 years) and 54 cognitively normal subjects (mean age: 74.3 +/- 8.1 years) were studied using MRSI and MRI. RESULTS: NAA concentration was less in patients with AD compared with healthy subjects by 21% (p < 0.0001) in the medial temporal lobe and by 13% to 18% (p < 0.003) in parietal lobe gray matter (GM), but was not changed significantly in white matter and frontal lobe GM. In addition to lower NAA, AD patients had 29% smaller hippocampi and 11% less cortical GM than healthy subjects. Classification of AD and healthy subjects increased significantly from 89% accuracy using hippocampal volume alone to 95% accuracy using hippocampal volume and NAA together. CONCLUSION: In addition to brain atrophy, NAA reductions occur in regions that are predominantly impacted by AD pathology. PMID- 11914411 TI - The value of sleep deprivation as a diagnostic tool in adult sleepwalkers. AB - OBJECTIVE: Adult somnambulism can result in injury to the sleeper and to others. Attempts to induce sleepwalking episodes in the sleep laboratory have yielded mixed results. Having shown that sleepwalkers have lower slow-wave activity power than control subjects, the authors hypothesized that an enhanced pressure of the homeostatic process underlying sleep regulation could affect the disorder's characteristics even further. Therefore, the effects of 38 hours of sleep deprivation in adult sleepwalkers and control subjects were investigated. METHODS: Ten adult somnambulistic patients and 10 sex- and age-matched control subjects were studied in the sleep laboratory. After a screening night, participants were monitored during 1) one night of baseline recording, and 2) one recovery night in which subjects slept ad libitum immediately after the sleep deprivation protocol. Behavioral manifestations were assessed for frequency and complexity using a 3-point scale of increasing complexity. RESULTS: None of the control subjects had any behavioral manifestations on either of the two nights. Conversely, sleepwalkers showed a significant increase in the frequency and complexity of the somnambulistic episodes during the recovery night compared with baseline. Somnambulistic patients had a greater number of awakenings from slow wave sleep than control subjects on both nights, but there was no significant increase during the recovery night. CONCLUSION: Sleep deprivation can be an effective tool for inducing somnambulistic episodes in the laboratory, thereby facilitating the diagnosis of sleepwalking. PMID- 11914412 TI - Absence of linkage to 8q24 in a European family with familial adult myoclonic epilepsy (FAME). AB - BACKGROUND: Familial adult myoclonic epilepsy (FAME) is defined by autosomal dominant inheritance, adult onset of myoclonus of the extremities, infrequent epileptic seizures, nonprogressive course, abnormality of polyspikes and waves on examination by EEG and photosensitivity, giant somatosensory evoked potentials, enhancement of C reflex, and premyoclonus spike detected by means of the jerk locked averaging EEG method. These findings were also observed in patients with benign adult familial myoclonic epilepsy (BAFME) and patients with familial cortical tremor. FAME and BAFME have been described only in Japan. The genes responsible for FAME and BAFME were mapped in the same genetic interval in 8q22.3 q24.1 OBJECTIVE: To study clinical and genetic characteristics of a European family with FAME. METHODS: A four-generation European kindred presenting with FAME, including 18 members, is described. Clinical analysis was performed on 15 living subjects and electrophysiologic study on 5 patients. Linkage analysis was performed with fluorescent microsatellites encompassing the FAME/BAFME locus (8q23.3-q24.1). RESULTS: Ten living and three deceased relatives had the clinical characteristics of FAME. Mean age at onset of the 10 living patients was 41 years (range, 30-60 years). Eight of the 13 affected subjects had generalized tonic clonic seizures. Electrophysiologic studies confirmed the diagnosis of FAME in the five patients studied. The pattern of inheritance was consistent with an autosomal dominant inheritance. The locus responsible for FAME/BAFME was excluded. CONCLUSION: Observation of a European family extends the occurrence of familial adult myoclonic epilepsy to non-Japanese patients. Exclusion of linkage of this family to the locus for familial adult myoclonic epilepsy/benign adult familial myoclonic epilepsy established the genetic heterogeneity of this disorder. PMID- 11914413 TI - Normobaric hyperoxia reduces MRI diffusion abnormalities and infarct size in experimental stroke. AB - BACKGROUND: Hyperbaric oxygen therapy is considered an important stroke treatment strategy. BACKGROUND: To determine whether normobaric oxygen is neuroprotective, and, if so, what the therapeutic time window is. METHODS: Experiment 1-Serial diffusion- and perfusion-weighted MRI (DWI and PWI) was performed after middle cerebral artery filament occlusion (MCAO) in rats randomized to FiO(2) 30% (normoxia) or FiO(2) 100% (hyperoxia). Experiment 2-48-hour lesion volumes were analyzed in rats subjected to 2-hour MCAO and randomized to normoxia or hyperoxia starting 15, 30, or 45 minutes after MCAO and ending 15 minutes after reperfusion. RESULTS: Experiment 1-Lesion apparent diffusion coefficient (ADC) values were persistently low in normoxic animals. In hyperoxia-treated rats, ADC values in cortical border zones showed progressive recovery from 66 +/- 3% of contralateral before hyperoxia, to 104 +/- 20% at approximately 2 hours. Striatal ADC values showed early but ill-sustained improvement. ADC lesion volumes increased progressively in the normoxia group. In the hyperoxia group, ADC lesion volumes tended to decrease after starting hyperoxia; however, lesions later increased in size, and 2-hour lesion volumes were not significantly different from baseline. PWI showed stable right MCA hypoperfusion in all animals. Experiment 2-Hyperoxia within 30 minutes significantly reduced total and cortical lesion volumes at 48 hours after stroke. Striatal lesion volumes were significantly reduced in the hyperoxia-15 group. CONCLUSION: In rats subjected to transient stroke, 100% oxygen administered within 30 minutes salvages ischemic brain tissue, especially in the cerebral cortex. Reducing the time to treatment enhances the degree of neuroprotection. PMID- 11914414 TI - The mystery of the Doctor's son, or the riddle of West syndrome. AB - Although the eponym "West syndrome" is used widely for infantile spasms, the originators of the term and the time frame of its initial use are not well known. This article provides historical details about Dr. West, about his son who had infantile spasms, and about the circumstances leading to the coining of the term West syndrome. PMID- 11914415 TI - Cycad neurotoxins, consumption of flying foxes, and ALS-PDC disease in Guam. AB - The Chamorro people of Guam have been afflicted with a complex of neurodegenerative diseases (now known as ALS-PDC) with similarities to ALS, AD, and PD at a far higher rate than other populations throughout the world. Chamorro consumption of flying foxes may have generated sufficiently high cumulative doses of plant neurotoxins to result in ALS-PDC neuropathologies, since the flying foxes forage on neurotoxic cycad seeds. PMID- 11914416 TI - "Economy class" stroke syndrome? AB - The authors report three cases of ischemic stroke in young adults that occurred during or after an airplane flight. Workup was negative for any cause of stroke other than the presence of a patent foramen ovale (PFO). There is an increasing awareness of deep vein thrombosis and pulmonary embolism occurring in relation to long flights. Individuals with a PFO under these circumstances may be vulnerable to stroke from paradoxic embolism. "Economy class" stroke syndrome may be underdiagnosed and is an eminently preventable cause of stroke. PMID- 11914417 TI - Progressive bulbospinal amyotrophy in triple A syndrome with AAAS gene mutation. AB - Triple A (3A) syndrome, a rare autosomal recessive disorder, is characterized by adrenocorticotropic hormone-resistant adrenal insufficiency, achalasia of the cardia, alacrima, and variable autonomic and neurologic dysfunction. The gene responsible, AAAS, recently has been identified. We describe the neurologic phenotype of the first adult case of 3A syndrome presenting bulbospinal amyotrophy as the prominent sign in association with a homozygous nonsense mutation identified in the AAAS gene. PMID- 11914418 TI - CAG/CTG repeat expansions at the Huntington's disease-like 2 locus are rare in Huntington's disease patients. AB - The authors report a large series of patients with Huntington disease (HD)-like phenotype without CAG repeat expansions in the IT15 gene that were screened for the newly identified CAG/CTG expansion in the gene encoding junctophilin-3. Normal alleles in controls had from 8 to 28 repeats. A single patient of North African origin with typical HD carried an allele with 50 uninterrupted repeats, representing approximately 2% of the non-IT15 HD patients tested. Therefore, further genetic heterogeneity is expected in HD. PMID- 11914419 TI - The cerebellum may be directly involved in cognitive functions. AB - Neuropsychological deficits may occur in infratentorial strokes. Only minor cognitive disturbances are reported in unilateral anterior cerebellar lesions. Here, the authors describe a patient with bilateral anterior ponto-cerebellar ischemic lesions associated with major neuropsychological deficits. Cerebral PET and SPECT demonstrated no metabolic defect in supratentorial areas. PMID- 11914420 TI - Midbrain deafness with normal brainstem auditory evoked potentials. AB - The authors report two cases of patients with word deafness. The word deafness occurred after a head injury for the first patient and after an arterio venous malformation embolization for the second patient. MRI demonstrated bilateral lesions of the inferior colliculi but brainstem auditory-evoked potentials (BAEP) were within normal limits. These cases demonstrated that lesions involving the two inferior colliculi induced pure word deafness but do not affect BAEP. PMID- 11914421 TI - HLA-A2 homozygosity but not heterozygosity is associated with Alzheimer disease. AB - AD is associated with the A2 allele of the human leukocyte antigen (HLA). However, it is not currently known whether there is any difference between A2 homozygotes and A2 heterozygotes. The authors studied 458 patients with AD and found that A2 homozygotes had earlier onset of AD than either A2 heterozygotes (5.4 years, p = 0.002) or those without A2 (5.2 years, p = 0.003). The "recessive" nature of this association suggests that loss of function at the HLA A locus or a closely linked gene is associated with AD. PMID- 11914422 TI - fMRI/EEG in paroxysmal activity elicited by elimination of central vision and fixation. AB - Using functional MRI (fMRI) with concurrent EEG monitoring the authors studied cortical activation associated with epileptiform discharges in three patients with fixation-off sensitivity. The EEG paroxysmal activity elicited by eliminating central vision correlated significantly with an increased blood oxygen level-dependent signal in the extrastriate cortex (Brodmann areas 19 and 37). fMRI provides a unique opportunity for localizing precisely the cortical areas generating paroxysmal activity in patients with fixation-off sensitivity. PMID- 11914423 TI - Chronic inflammatory demyelinating polyneuropathy presenting with features of GBS. AB - The authors report five patients with inflammatory demyelinating polyneuropathy with a Guillain-Barre syndrome (GBS)-like onset and initial clinical features, but with persistent symptoms similar to chronic inflammatory demyelinating polyneuropathy (CIDP). Patients in the chronic phase improved with corticosteroid or IV immunoglobulin therapy. Patients with apparent GBS who show persistent symptoms may benefit from corticosteroids or other treatment that is beneficial in the management of CIDP. PMID- 11914424 TI - Spinocerebellar ataxia type 10 is rare in populations other than Mexicans. PMID- 11914425 TI - Multifocal myoclonus due to verapamil overdose. PMID- 11914426 TI - Isolated weakness of index finger due to small cortical infarction. PMID- 11914427 TI - Intraluminal carotid thrombus. PMID- 11914428 TI - Intention tremor, parkinsonism, and generalized brain atrophy in male carriers of fragile X. PMID- 11914429 TI - Ring chromosome 20 epilepsy syndrome in children: electroclinical features. PMID- 11914430 TI - CSF filtration is an effective treatment of Guillain-Barre syndrome: a randomized clinical trial. PMID- 11914431 TI - Accuracy of clinical criteria for AD in the Honolulu-Asia Aging Study, a population-based study. PMID- 11914432 TI - How many patients with psychogenic nonepileptic seizures also have epilepsy? PMID- 11914433 TI - Limb immobilization for the treatment of focal occupational dystonia. PMID- 11914435 TI - Covariation of psychosocial characteristics associated with cardiovascular disease: genetic and environmental influences. AB - OBJECTIVE: Three psychosocial characteristics associated with cardiovascular disease (CVD)-depression, hostility, and social support-tend to correlate with one another. However, the causes of each characteristic and why they tend to co occur are not completely understood. Therefore, the current study used a twin design to examine the relative contributions of genetic and environmental influences to the variation and covariation of these three psychosocial characteristics. METHODS: The sources of variation and covariation among the Beck Depression Inventory, the Cook-Medley Hostility Scale, and the Interpersonal Support Evaluation List were examined in a young adult community sample of 157 monozygotic and 75 dizygotic twin pairs. RESULTS: Phenotypic confirmatory factor analysis indicated that a single latent factor could account for their moderate intercorrelations. Twin analyses indicated that the Beck Depression Inventory and Interpersonal Support Evaluation List were each influenced by genetic and nonshared environmental factors, whereas the Cook-Medley Hostility Scale was influenced by familial (genetic and/or shared environmental) and nonshared environmental factors. Bivariate associations between these scales were largely determined by common genetic effects and, to a lesser degree, common nonshared environmental effects. Covariation among the three scales could be explained by a single common genetic factor and a common nonshared environmental factor. Environmental factors shared within families did not contribute to covariation among the psychosocial characteristics. CONCLUSIONS: The results challenge the conventional approach of examining these psychosocial variables as independent risk factors for cardiovascular disease and argue for the importance of investigating specific causes for their covariation. PMID- 11914437 TI - History of psycho-oncology: overcoming attitudinal and conceptual barriers. AB - The formal beginnings of psycho-oncology date to the mid-1970s, when the stigma making the word "cancer" unspeakable was diminished to the point that the diagnosis could be revealed and the feelings of patients about their illness could be explored for the first time. However, a second stigma has contributed to the late development of interest in the psychological dimensions of cancer: negative attitudes attached to mental illness and psychological problems, even in the context of medical illness. It is important to understand these historical underpinnings because they continue to color contemporary attitudes and beliefs about cancer and its psychiatric comorbidity and psychosocial problems. Over the last quarter of the past century, psycho-oncology became a subspecialty of oncology with its own body of knowledge contributing to cancer care. In the new millennium, a significant base of literature, training programs, and a broad research agenda have evolved with applications at all points on the cancer continuum: behavioral research in changing lifestyle and habits to reduce cancer risk; study of behaviors and attitudes to ensure early detection; study of psychological issues related to genetic risk and testing; symptom control (anxiety, depression, delirium, pain, and fatigue) during active treatment; management of psychological sequelae in cancer survivors; and management of the psychological aspects of palliative and end-of-life care. Links between psychological and physiological domains of relevance to cancer risk and survival are being actively explored through psychoneuroimmunology. Research in these areas will occupy the research agenda for the first quarter of the new century. At the start of the third millennium, psycho-oncology has come of age as one of the youngest subspecialties of oncology, as one of the most clearly defined subspecialties of consultation-liaison psychiatry, and as an example of the value of a broad multidisciplinary application of the behavioral and social sciences. PMID- 11914438 TI - Posttraumatic stress disorder revisited. AB - In this review we trace the history of and professional discussion on psychological traumatization due to "railroad spine syndrome," "shell-shock syndrome," and "war neuroses," as well as the more or less endemic "posttraumatic stress disorder" of today. Psychological trauma engenders longlasting consequences in the biological, intrapsychic, and social organization of individuals. Medical experts have reported a shift in attention from exogenous to endogenous and back to exogenous causes, as indicated by new diagnostic systems (DSM-IV and ICD-10). As far as the relevant literature is concerned, the medical profession demonstrates the same partial amnesia as their patient counterparts. The purpose of this review is to overcome this fragmented memory and thus reach a more integrated view of what constitutes psychological trauma by reviewing trauma related articles published in Psychosomatic Medicine. Moreover, we point out the direction in which research is desperately needed and ought to develop. PMID- 11914439 TI - Marked lability in urinary cortisol levels in subgroups of combat veterans with posttraumatic stress disorder during an intensive exposure treatment program. AB - OBJECTIVE: The objective of this study was to obtain longitudinal data on lability of cortisol levels in posttraumatic stress disorder (PTSD) because previous studies have largely been based on sampling at a single time point and have yielded varying results. METHODS: This study measured urinary cortisol levels at admission, midcourse, and discharge during a 90-day hospitalization period in male Vietnam combat veterans with PTSD (N = 51). RESULTS: Although there were no significant differences in the mean +/- SEM urinary cortisol levels between the admission (59.4 +/- 3.0 microg/d), midcourse (55.6 +/- 3.9 microg/d), and discharge (53.4 +/- 3.4 microg/d) values, marked lability of cortisol levels in individual patients was observed over time, with changes ranging from +93 to 58 microg/d from admission to midcourse. In addition, this hormonal lability defined discrete subgroups of patients on the basis of the longitudinal pattern of cortisol change during exposure treatment, and there were significant psychometric differences in the level of social functioning between these subgroups. CONCLUSIONS: The findings do not support the concept of either a static "hypocortisolism" or "hypercortisolism" in PTSD, but rather suggest a psychogenic basis for cortisol alterations in PTSD in relation to psychosocial stress and indicate a central regulatory dysfunction of the hypothalamic pituitary-adrenal axis characterized by a dynamic tendency to overreact in both upward and downward directions. The longitudinal findings fit with recent observations that cortisol elevations occur when acutely superimposed stressful conditions emotionally engage patients and overwhelm the usually dominating disengaging coping mechanisms associated with suppression of cortisol levels in PTSD. The findings emphasize the importance of longitudinal data in studies of the hypothalamic-pituitary-adrenal axis in PTSD. PMID- 11914440 TI - Relationship of clinic, ambulatory, and laboratory stress blood pressure to left ventricular mass in overweight men and women with high blood pressure. AB - OBJECTIVE: This study was designed to evaluate the relationship between left ventricular (LV) mass and blood pressure (BP) recorded in the following contexts: in the clinic, using standard auscultatory procedures, during a typical day using ambulatory BP monitoring, and in the laboratory environment during behavioral stress testing. METHODS: Ninety-seven men and women with clinic systolic blood pressure (SBP) of 130 to 180 mm Hg and/or diastolic blood pressure (DBP) of 85 to 110 mm Hg and mild to moderate obesity were included in the study. Laboratory stressors included the following tasks: Public Speaking; Anger Interview; Mirror Trace; and Cold Pressor. LV mass was measured using echocardiography and adjusted for body size by dividing by height(2.7) to yield LV mass index (LVMI). RESULTS: LVMI was positively correlated with clinic SBP (r = 0.24, p <.05), ambulatory SBP (r = 0.34, p <.01), and aggregated laboratory stress SBP (r = 0.28, p <.01). Of the individual stressors, only SBP responses to the Mirror Trace and Cold Pressor tasks were independently correlated with LVMI (r = 0.35 and 0.34, respectively, p values <.01). Hierarchical regression analyses revealed that laboratory stress SBP remained a significant predictor of LVMI, after controlling for BMI and clinic pressure. CONCLUSIONS: These findings suggest that cardiovascular responses to behavioral stress are associated with individual differences in LVMI in men and women with high blood pressure who are overweight. Laboratory studies of behavioral stress may help promote our understanding of the pathophysiology of LVH. PMID- 11914441 TI - The PHQ-15: validity of a new measure for evaluating the severity of somatic symptoms. AB - OBJECTIVE: Somatization is prevalent in primary care and is associated with substantial functional impairment and healthcare utilization. However, instruments for identifying and monitoring somatic symptoms are few in number and not widely used. Therefore, we examined the validity of a brief measure of the severity of somatic symptoms. METHODS: The Patient Health Questionnaire (PHQ) is a self-administered version of the PRIME-MD diagnostic instrument for common mental disorders. The PHQ-15 comprises 15 somatic symptoms from the PHQ, each symptom scored from 0 ("not bothered at all") to 2 ("bothered a lot"). The PHQ-15 was administered to 6000 patients in eight general internal medicine and family practice clinics and seven obstetrics-gynecology clinics. Outcomes included functional status as assessed by the 20-item Short-Form General Health Survey (SF 20), self-reported sick days and clinic visits, and symptom-related difficulty. RESULTS: As PHQ-15 somatic symptom severity increased, there was a substantial stepwise decrement in functional status on all six SF-20 subscales. Also, symptom related difficulty, sick days, and healthcare utilization increased. PHQ-15 scores of 5, 10, 15, represented cutoff points for low, medium, and high somatic symptom severity, respectively. Somatic and depressive symptom severity had differential effects on outcomes. Results were similar in the primary care and obstetrics-gynecology samples. CONCLUSIONS: The PHQ-15 is a brief, self administered questionnaire that may be useful in screening for somatization and in monitoring somatic symptom severity in clinical practice and research. PMID- 11914442 TI - Reduced primary antibody responses in a genetic animal model of depression. AB - OBJECTIVE: Clinical depression is associated with multiple abnormalities of immune function, including reduced virus-specific responses. This study tested the hypothesis that the Flinders Sensitive Line (FSL) rat, a promising genetic animal model of depression, would exhibit reductions in antigen-specific primary antibody responses to immunization. METHODS: FSL (N = 13) and control Flinders Resistant Line (FRL; N = 14) rats were immunized with the protein antigen keyhole limpet hemocyanin (KLH; 300 microg/kg), and KLH-specific immunoglobulin (Ig)M, IgG, IgG1, and IgG2a responses were measured before and 3, 5, 7, 11, and 14 days after immunization. In separate experiments, production of interferon-gamma (IFN gamma) by cells from naive and KLH-immunized animals was measured in vitro to determine whether strain differences in antibody production might be associated with differential production of this regulatory cytokine. RESULTS: KLH-specific production of IgM (p <.01) and IgG2a (p <.05) was significantly reduced in the FSL rats compared with the FRL controls. There were no strain differences in IgG or IgG1 production. Although IFN-gamma production between the two strains was similar in naive animals, cells from KLH-immunized FSL rats produced significantly less IFN-gamma when stimulated with KLH in vitro than cells from KLH-immunized FRL controls (p =.01). CONCLUSIONS: This study extends previous reports of altered immune function in the FSL rats to include reduced in vivo antigen-specific antibody responses. Moreover, diminished production of IFN-gamma by KLH-primed lymphocytes may contribute to lower antibody production in these animals. Collectively, these data suggest deficiencies in type 1 T-helper cell mediated immunity in the FSL rats. PMID- 11914443 TI - Hypercortisolemic depression is associated with increased intra-abdominal fat. AB - OBJECTIVE: Similar to patients with a metabolic syndrome, patients with major depression are at increased risk of developing cardiovascular disorders. Interestingly, both disorders share a specific endocrine syndrome that promotes the accumulation of visceral fat, which again is considered a marker of increased cardiovascular morbidity and mortality. METHODS: Intra-abdominal fat was measured in 22 postmenopausal depressed women and 23 age-matched healthy women by computer tomography at the level of lumbar vertebrae 1 (L1) and 4 (L4). Saliva was taken in patients and control subjects at 08:00 hours over a period of 7 drug-free days for the measurement of free cortisol. In patients only we performed an oral glucose tolerance test. RESULTS: Compared with control subjects, depressed patients with elevated free cortisol concentrations showed similar visceral fat depots at L1 (113.0 +/- 41.6 vs. 94.3 +/- 53.2 cm(2)). Hypercortisolemic depressed patients also showed greater fat depots in this area (74.5 +/- 55.5 cm(2), p =.04) than the normocortisolemic patients. However, a comparison of all patients with control subjects revealed no difference in fat accumulation at either L1 or L4. Finally, glucose concentrations during the glucose tolerance test were higher in hypercortisolemic than in normocortisolemic patients, whereas their insulin levels showed only a tendency toward being increased. CONCLUSIONS: Hypercortisolemic depressed patients suffer from resistance to insulin and increased visceral fat. The fact that hypercortisolemia reverses depression related fat loss, particularly in the visceral area, might partially explain why major depression can be considered a risk factor for cardiovascular disorders. PMID- 11914444 TI - A novel stress and coping workplace program reduces illness and healthcare utilization. AB - OBJECTIVE: The purpose of this study was to determine if a novel workplace stress management program, delivered either face-to-face or by self-help, would reduce illness and health services utilization among participants. METHODS: Five hundred one volunteers were randomly allocated to one of three groups: full intervention, which received assessment and personalized self-study feedback and was offered six face-to-face, small-group sessions; partial intervention, a self-help group that received assessment and personalized feedback by mail; and a wait-list control group. All participants completed questionnaires for stress, anxiety, and coping at the start of the study and 6 and 12 months later. Health reports were completed at 0, 3, 6, 9, and 12 months. A subsample of subjects who subscribed to a single health maintenance organization provided objectively recorded doctor visit data across the study year. RESULTS: All three groups reported significant improvement in their stress, anxiety, and coping across the year. Full intervention participants showed a more rapid reduction in negative responses to stress than did participants from the other groups. Full-intervention subjects also reported fewer days of illness than subjects in the other groups. Objectively measured physician visits showed a large (34%) reduction in healthcare utilization for full intervention subjects in the HMO subsample. CONCLUSIONS: These results indicated that a work-site program that focuses on stress, anxiety, and coping measurement along with small-group educational intervention can significantly reduce illness and healthcare utilization. PMID- 11914445 TI - Stress, coping, and hepatitis B antibody status. AB - OBJECTIVE: The present study investigated the association between exposure to stressful life events, coping style, and antibody status after hepatitis B vaccination. METHODS: Two hundred sixty medical school undergraduates, who had received the three-dose hepatitis B vaccine before recruitment to this study, completed questionnaires measuring exposure to stressful life events during the past year, customary coping strategies, and health behaviors. Antibodies against hepatitis B surface antigen were determined; levels <100 mIU/ml were deemed inadequate. RESULTS: Two participant cohorts were identified: those vaccinated within the last year and those vaccinated earlier. In the early vaccination cohort, participants with greater-than-average stress exposures had a more than two-fold increased risk of having an inadequate antibody titer. Coping by accepting the reality of stressful situations proved protective, whereas coping by substance use increased the risk of having an inadequate antibody count in this cohort. These associations remained significant after adjustment for possible mediators. Furthermore, the effects of stress and coping were largely independent of one another. Neither stress nor coping was significantly associated with antibody status in the recently vaccinated cohort. CONCLUSIONS: The present study confirms that the immune system is sensitive to variations in psychological factors. Stressful life events and coping strategy seem to have a continuing impact on hepatitis B antibody status. PMID- 11914447 TI - Reactivity and vulnerability to stress-associated risk for upper respiratory illness. AB - OBJECTIVE: We tested the hypothesis that the greater a person's laboratory stress elicited elevation in cortisol, the greater the life stress-related risk for upper respiratory infection (URI). We also tested the prediction that the greater the laboratory stress-elicited rise in natural killer cell (NK) cytotoxicity, the smaller the life stress-related URI risk. Finally, we explored whether sympathetic nervous system (SNS) and enumerative immune reactivities to laboratory stress moderate the relation between life stress and URI. METHODS: At baseline, 115 healthy subjects were administered a negative stressful life events checklist and were tested to assess their SNS (blood pressure, heart rate, and catecholamines), HPA (cortisol), and immune (NK cell cytotoxicity and lymphocyte subsets) reactivities to laboratory speech tasks administered 2 weeks apart. Responses were averaged across the two laboratory assessments to create reactivity scores. After these assessments were completed, participants were followed weekly for 12 consecutive weeks. At each follow-up they completed a measure of perceived stress experienced over the last week. They were also instructed to contact the study coordinator if they had a cold or flu at any time during follow-up. A health care worker verified reported illnesses. RESULTS: In a traditional prospective analysis, high cortisol reactors with high levels of life events had a greater incidence of verified URI than did high reactors with low levels of life events and low reactors irrespective of their life event scores. Using hierarchical linear modeling, CD8(+) number, Natural Killer (NK) cell number, and NK cell cytotoxicity, each interacted with weekly perceived stress levels in predicting concurrent occurrences of self-reported URIs. For these outcomes, low immune reactors were more likely to experience an URI during high stress than low stress weeks. High immune reactors did not exhibit differences in weekly URIs as a function of weekly stress level. The SNS reactivity markers did not moderate the association of stress and URI incidence in either analysis. CONCLUSIONS: Acute HPA and immune responses to laboratory stressors are markers of how vulnerable people are to the increased risk for URI associated with stressors in the natural environment. PMID- 11914446 TI - Defensiveness and individual response stereotypy in asthma. AB - OBJECTIVE: Previous literature has shown that the psychological trait of defensiveness is related to elevated sympathetic reactivity to stress and to several cardiac risk factors. The aim of this study was to examine whether these previous findings on defensiveness extend to an asthmatic population. METHODS: Defensiveness was measured by the Marlowe-Crowne Social Desirability Scale using a quartile split: high (upper 25%) and low (bottom 75%). Twenty-two defensive and 66 nondefensive participants with asthma were exposed to laboratory tasks (initial baseline rest period, reaction time task, and a shop accident film). RESULTS: During the tasks there was evidence of lower skin conductance levels and greater respiratory sinus arrhythmia amplitudes among defensive patients with asthma. After exposure to the tasks, defensive patients with asthma showed a decline on spirometry test measures compared with nondefensive asthmatic patients, who displayed an increase. CONCLUSIONS: These data confirm individual response stereotypy and suggest that defensiveness may be characterized by sympathetic hypoarousal and parasympathetic hyperarousal among patients with asthma. Future studies are needed to determine whether defensiveness is a risk factor for stress-induced bronchoconstriction. PMID- 11914448 TI - Low-dose dexamethasone suppression test in chronic fatigue syndrome and health. AB - OBJECTIVE: Subtle dysregulations of the hypothalamus-pituitary-adrenal axis in chronic fatigue syndrome have been described. The aim of this study was to examine the negative feedback regulations of the hypothalamus-pituitary-adrenal axis in chronic fatigue syndrome. METHODS: In 21 patients with chronic fatigue syndrome and 21 healthy control subjects, awakening and circadian salivary free cortisol profiles were assessed over 2 consecutive days and compared with awakening and circadian salivary free cortisol profiles after administration of 0.5 mg of dexamethasone at 11:00 PM the previous day. RESULTS: Patients with chronic fatigue syndrome had normal salivary free cortisol profiles but showed enhanced and prolonged suppression of salivary free cortisol after the administration of 0.5 mg of dexamethasone in comparison to the control subjects. CONCLUSIONS: Enhanced negative feedback of the hypothalamus-pituitary-adrenal axis could be a plausible explanation for the previously described alterations in hypothalamus-pituitary-adrenal axis functioning in chronic fatigue syndrome. Because similar changes have been described in stress-related disorders, a putative role of stress in the pathogenesis of the enhanced feedback is possible. PMID- 11914449 TI - Prognosis of chronic fatigue in a community-based sample. AB - OBJECTIVE: This study examined predictors of fatigue severity and predictors of continued chronic fatigue status at wave 2 follow-up within a random, community based sample of individuals previously evaluated in a wave 1 prevalence study of chronic fatigue and chronic fatigue syndrome that originally took place between 1995 and 1997. METHODS: Wave 1 data were from a larger community-based prevalence study of chronic fatigue syndrome. In the present study, a second wave of data were collected by randomly selecting a sample of participants from the wave 1 sample of 18,675 adults and readministering a telephone screening questionnaire designed to assess symptoms of chronic fatigue syndrome. RESULTS: Findings revealed that wave 1 fatigue severity was a predictor of fatigue severity at wave 2 in the overall sample of individuals with and without chronic fatigue. In the smaller sample of individuals with chronic fatigue, wave 1 fatigue severity, worsening of fatigue with physical exertion, and feeling worse for 24 hours or more after exercise significantly predicted continued chronic fatigue status (vs. improvement) at wave 2 follow-up. CONCLUSIONS: These findings underscore the prognostic validity of postexertional malaise in predicting long-term chronic fatigue and also highlight the importance of using population-based, representative random samples when attempting to identify long-term predictors of chronic fatigue at follow-up. PMID- 11914450 TI - Atypical sexual behavior during sleep. AB - OBJECTIVE: This article reports a case series of atypical sexual behavior during sleep, which is often harmful to patients or bed partners. METHODS: Eleven subjects underwent clinical evaluation of complaints of sleep-related atypical sexual behavior. Complaints included violent masturbation, sexual assaults, and continuous (and loud) sexual vocalizations during sleep. One case was a medical legal case. Sleep logs, clinical evaluations, sleep questionnaires, structured psychiatric interviews, polysomnography, actigraphy, home electroencephalographic monitoring during sleep, and clinical electroencephalographic monitoring while awake and asleep were used to determine clinical diagnoses. RESULTS: Atypical sexual behaviors during sleep were associated with feelings of guilt, shame, and depression. Because of these feelings, patients and bed partners often tolerated the abnormal behavior for long periods of time without seeking medical attention. The following pathologic sleep disorders were demonstrated on polysomnography: partial complex seizures, sleep-disordered breathing, stage 3 to 4 non-rapid eye movement (REM) sleep parasomnias, and REM sleep behavior disorder. These findings were concurrent with morning amnesia. CONCLUSIONS: The atypical behaviors were related to different syndromes despite the similarity of complaints from bed partners. In most cases the disturbing and often harmful symptoms were controlled when counseling was instituted and sleep disorders were treated. In some cases treatment of seizures or psychiatric disorders was also needed. Clonazepam with simultaneous psychotherapy was the most common successful treatment combination. The addition of antidepressant or antiepileptic medications was required in specific cases. PMID- 11914451 TI - Socioeconomic status and health: the role of sleep. AB - OBJECTIVE: Examine the role of sleep in the relationship between socioeconomic status (SES) and health. METHOD: Self-reported measures of income and education, sleep quantity and quality, and mental and physical health were obtained in a community sample of 1139 adults. RESULTS: More education was associated with higher income (p <.001), and higher income was associated with better physical health (p <.001) and psychological outcomes (p <.001). The effects of income on both mental and physical health were mediated by sleep quality (p values <.01), and sleep quantity was related to both measures of health (p values <.01) but to neither index of SES. CONCLUSION: Sleep quality may play a mediating role in translating SES into mental and physical well-being, and income seems to mediate the effect of education on sleep and, in turn, health. PMID- 11914452 TI - Critical incident exposure and sleep quality in police officers. AB - OBJECTIVE: Police officers face many stressors that may negatively impact sleep quality. This study compares subjective sleep quality in police officers with that in control subjects not involved in police or emergency services. We examined the effects of critical incident exposure (trauma exposure) and routine (nontraumatic) work environment stressors on sleep quality after controlling for the effects of work shift schedule. METHODS: Subjective sleep disturbances were measured by the Pittsburgh Sleep Quality Index in police officers (variable-shift workers, N = 551; stable day-shift workers, N = 182) and peer-nominated comparison subjects (variable-shift workers, N = 98; stable day-shift workers, N = 232). The main predictor variables were 1) duty-related critical incident exposure to on-line policing and 2) work environment stress related to routine administrative and organizational aspects of police work. RESULTS: Police officers on both variable and stable day shifts reported significantly worse sleep quality and less average sleep time than the two corresponding control groups. Within police officers, cumulative critical incident exposure was associated with nightmares but only weakly associated with poor global sleep quality. In contrast, the stress from officers' general work environment was strongly associated with poor global sleep quality. Sleep disturbances were strongly associated with posttraumatic stress symptoms and general psychopathology. CONCLUSIONS: A large percentage of police officers report disturbances in subjective sleep quality. Although the life-threatening aspects of police work are related to nightmares, the routine stressors of police service seem to most affect global sleep quality in these subjects. These findings may have implications for health and occupational performance. PMID- 11914453 TI - Somatosensory amplification and its relationship to heartbeat detection ability. AB - OBJECTIVE: Somatosensory amplification has been defined as the tendency to experience normal bodily sensations as intense, noxious, and disturbing. The present experiment investigated whether this tendency is due to heightened physiological sensitivity to bodily sensations. METHODS: The relationship between Somatosensory Amplification Scale (SSAS) scores and objective measures of the ability to detect bodily (ie, heartbeat) sensations derived from the Method of Constant Stimuli procedure was assessed. Although somatosensory amplification characterizes hypochondriacs, the relationship between somatosensory amplification and sensitivity to bodily sensations was examined in nonhypochondriacal, nonpatient participants in an effort to dissociate somatosensory amplification from other variables associated with hypochondriasis and/or patient status. RESULTS: Heartbeat detectors were found to exhibit significantly lower SSAS scores than nondetectors. CONCLUSION: This finding suggests that somatosensory amplification is not due to heightened sensitivity to bodily sensations. PMID- 11914454 TI - Patients with erythrophobia (fear of blushing) show abnormal autonomic regulation in mental stress conditions. AB - OBJECTIVE: The objective of this study was to analyze the autonomic functions of patients with erythrophobia. METHODS: Forty patients with a diagnosis of erythrophobia (female/male ratio 18/22) without any other organic lesions and 20 healthy volunteers (female/male ratio 10/10) were assessed. Clinical evaluation was performed using a modified version of semistructured interviews. Autonomic testing was performed by means of spectral analysis of heart rate and continuous blood pressure by sparse discrete Fourier transformation at rest and under mental stress. RESULTS: There were no significant difference between the two samples in age, sex distribution, BMI, resting systolic, or diastolic blood pressure, nor was there a difference in autonomic baseline functioning between the 40 patients with erythrophobia and the control subjects. On the other hand, patients with erythrophobia consistently showed higher pulse rates (88 +/- 20 vs. 78 +/- 9 bpm, p <.05), higher total heart rate power values (8.40 +/- 0.63 vs. 8.07 +/- 1.02 p <.05), higher midfrequency spectral values (7.38 +/- 0.66 vs. 7.02 +/- 1.18, p <.01), higher high-frequency spectral values (6.89 +/- 0.86 vs. 6.48 +/- 1.44, p <.05), and lower baroreceptor sensitivity (8.62 +/- 8.16 vs. 11.65 +/- 4.42, p <.005) than the healthy subjects. ANOVA showed a significant group interaction (p <.0001) between the samples. CONCLUSIONS: This study provides evidence for abnormal autonomic functioning in patients with erythrophobia when under mental stress. PMID- 11914456 TI - Declaration of the rights of older persons. PMID- 11914455 TI - In memoriam: Bernice L. Neugarten. PMID- 11914457 TI - Yali's question and the study of nursing homes as organizations. PMID- 11914459 TI - An exploration of job, organizational, and environmental factors associated with high and low nursing assistant turnover. AB - PURPOSE: This article examines factors that distinguish nursing facilities with very high and very low nursing assistant turnover rates from a middle referent group, exploring the possibility that high and low turnover are discrete phenomena with different antecedents. DESIGN AND METHODS: Data from a stratified sample of facilities in eight states, with directors of nursing as respondents (N = 288), were merged with facility-level indicators from the On-Line Survey Certification of Automated Records and county-level data from the Area Resource File. Multinominal logistic regression was used to identify factors associated with low (less than 6.6% in 6 months) and high (more than 64% in 6 months) turnover rates. RESULTS: With the exception of registered nurse turnover rate, low turnover and high turnover were not associated with the same factors. IMPLICATIONS: Future studies of facility turnover should avoid modeling turnover as a linear function of a single set of predictors in order to provide clearer recommendations for practice. PMID- 11914461 TI - Patterns in functioning among residents of an affordable assisted living housing facility. AB - PURPOSE: This is the second article reporting a project that (a) characterized the longitudinal functional patterns of residents in an affordable, assisted living housing (ALH) milieu for low-income people and (b) compared the ALH residents' functional patterns with those for similarly low-income, community dwelling people. DESIGN AND METHODS: Functioning was defined as the ability to perform activities of daily living and instrumental activities of daily living and respondents' competing risks such as death and institutionalization. The comparison samples were selected from the Health and Retirement Study (HRS) and the Asset and Health Dynamics Among the Oldest Old (AHEAD) study. The analyses involved description of functional patterns and multinomial logit models to assess the relative likelihood that study participants experienced each pattern of functioning and competing risks. RESULTS: About 42.4% of the ALH residents had stable high functioning, 32.9% deteriorated (e.g., had increased impairment, institutionalization, death), 3% improved in functioning, 6% exhibited moderate to severe impairment throughout the study, and the remainder returned to the community. Comparison of the ALH residents' functional patterns with those of the community-dwelling respondents indicated that the ALH residents' patterns generally were similar and in some ways better, adjusting for social-demographic factors and health; for example, the ALH residents were no different (statistically) in terms of improvement, decline, and death, but they were more likely to have stable high functioning. IMPLICATIONS: The results suggest that the affordable ALH milieu enabled low-income residents to maintain function or cope well with functional decline. The comparison with community-dwelling participants provides evidence that the affordable ALH milieu was comparatively beneficial. PMID- 11914460 TI - Mental health correlates of aggression in nursing home residents with dementia. AB - PURPOSE: Aggression continues to challenge caregivers of persons with dementia, and identification of foci for effective interventions is needed. The purpose of this study was to examine the influence of (a) the resident characteristics of depression, communication, and cognition and (b) behavior management strategies on aggression in a group of older nursing home residents (N = 405) with evidence of dementia. DESIGN AND METHODS: This cross-sectional correlational study examined the association of resident characteristics and behavior management strategies with bivariate aggression, physical aggression, and verbal aggression, using hierarchical regression. RESULTS: Main findings are that impaired communication is associated with all forms of aggression, depression with physical aggression, and disorientation with verbal aggression. A 3-month prior pattern of antipsychotic drug use was consistently associated with all forms of aggression and physical restraint use with physical aggression. Most of the explained variance was attributable to antipsychotic drug use. IMPLICATIONS: The separate explanatory models for physical and verbal aggression indicate that these may be unique entities with different foci for treatment. These results provide evidence that aggression persists despite antipsychotic drug use and that further mental health interventions might be targeted at compensating for impaired communication and the treatment of depression to improve the mental health of nursing home residents with dementia. PMID- 11914462 TI - Resident satisfaction and its components in residential aged care. AB - PURPOSE: The purpose of this study was to assess the direction and magnitude of the effects among the components of resident satisfaction in residential aged care and to examine if the relationships among satisfaction components vary according to facility type (i.e., nursing home and hostel). Briefly, a hostel is a low-care facility in which residents are more independent and receive personal but not nursing care. DESIGN AND METHODS: A cross-sectional survey design was adopted to collect the required information, and a stratified random sampling approach was used to select facilities. Structural equation modeling was used to examine relationships among satisfaction components in a sample of 394 nursing home and 752 hostel residents. RESULTS: The results indicate that satisfaction with staff care has a moderate and positive effect on all other aspects of resident satisfaction. The relationships among satisfaction components are different for nursing home and hostel residents. IMPLICATIONS: The findings lead to an improved understanding of the interrelationship among resident satisfaction components, which has important implications for improving quality outcomes (e.g., resident satisfaction) through appropriate intervention (e.g., enhancing staff care). PMID- 11914463 TI - Profile of home care aides, nursing home aides, and hospital aides: historical changes and data recommendations. AB - PURPOSE: To examine demographic characteristics and work conditions of home care aides, nursing home aides, and hospital aides in the late 1980s and late 1990s. DESIGN AND METHODS: This study replicated a previous study which examined the Current Population Survey (CPS) March supplement from 1987 to 1989. The present study examined CPS data from 1997 to 1999 both to obtain a profile of aides in the late 1990s and to compare 1990's work conditions and demographic characteristics with those of aides in the late 1980s. RESULTS: Whereas demographic characteristics and work conditions of hospital and nursing home aides show little change over the decade, home care aides today are generally younger, more educated, more likely to hold full-time positions, and more likely to have health benefits than their counterparts 10 years ago. Yet work conditions are still poorer for home care aides than for the other types of aides. IMPLICATIONS: Work conditions need further improvement, particularly for home care aides. More accurate data to describe paraprofessional care workers are also necessary to address the serious worker shortage problems. PMID- 11914464 TI - Health care consumption among elderly patients in california: a comprehensive 10 year evaluation of trends in hospitalization rates and charges. AB - PURPOSE: This report examines health care rates, charges, and patterns of consumption from a comprehensive California hospitalization data set covering 1986-1995. An improved understanding of current trends in health care consumption would facilitate the development of future resource allocation models. DESIGN AND METHODS: We obtained discharge and charge data from all licensed nonfederal hospitals in California between 1986 and 1995 relating to inpatient discharges of individuals aged 55 years and older. We used the direct method of standardization to adjust discharge statistics for differing age and gender case mixes, and we adjusted all charges to 1990 dollars for cost comparisons. RESULTS: Standardized to the 1990 population, annual discharge rates declined between 1986 and 1992, then leveled off to about 227 per 1,000 between 1993 and 1995. Rates of both discharges and charges for men consistently exceeded those for women, there being about a 5-year lag between female and male rates of discharge. The insurance payer mix shifted between 1986 and 1995, with dramatic declines in private insurance mirrored by increases in managed care. IMPLICATIONS: Hospital care consumption among the elderly people in California demonstrates a trend of increasing adjusted total charges despite declining hospitalization rates. Overall, individuals aged 55 years and older comprise 18% of the California population and incur 52% of discounted total charges. Private insurance has virtually disappeared, replaced by HMO/PHP/PPO organizations; still, charges to governmental sources (primarily Medicare and Medi-Cal) account for about 78% of total billings. Absolute numbers of Californians aged 55 and older are projected to increase 54% by 2010 and 226% by 2025 compared with 1995, engendering a dramatic increase in the financial burden of health care to this segment of the population. PMID- 11914465 TI - Continuing and ceasing leisure activities in later life: a longitudinal study. AB - PURPOSE: This study examined changes in leisure activities of older adults over an 8-year period, and associated sociodemographic and health characteristics. DESIGN AND METHODS: Data were from a longitudinal study conducted in Winnipeg, Manitoba, Canada; 380 respondents were interviewed in-person in both 1985 and 1993. Changes in ten specific activities and the overall number of activities continued were examined. RESULTS: Theater/movies/spectator sports and travel were the activities least likely to be continued over the 8-year period; watching television and reading were most likely to be continued. Characteristics significantly related to changes in activities were age, gender, education, and self-rated health in 1985 as well as changes in marital status, self-rated health, and functional ability between 1985 and 1993, although no consistent pattern emerged. IMPLICATIONS: Leisure education is discussed as a means of introducing modifications to enhance older adults' participation in desired activities. Directions for future research are highlighted. PMID- 11914466 TI - Systems of social support in families who care for dependent African American elders. AB - PURPOSE: This study examined connections (linking, compensatory, or none) between three systems of social support (informal, church, and formal). Predictors of each system were also examined. DESIGN AND METHODS: A community sample of 187 caregivers who provided care to older African American participants in the Duke Established Populations for Epidemiological Studies of the Elderly was used. Multiple regression analyses were used to examine relationships between systems of social support as well as to determine predictors of each social support system. RESULTS: Findings provide evidence for linking connections between systems of support: Informal and formal supports were linked, and church support and formal support were linked. The relationship of need as positively related to social support was limited to the care recipient's number of instrumental activities of daily living limitations being associated with formal support. Level of education was also associated with use of formal support. IMPLICATIONS: Cohesive family networks and network size are important factors that help determine what support caregivers receive. African American caregivers may be at risk for negative health outcomes because they are less likely to use formal support as care recipients' activities of daily living limitations increase. PMID- 11914467 TI - Issues of race, ethnicity, and culture in caregiving research: a 20-year review (1980-2000). AB - PURPOSE: The goals of this review are to synthesize what is known about caregiving among diverse groups and to identify gaps in knowledge to guide future research on caregiving. The review focuses on conceptual and theoretical approaches, sampling strategies, measurement techniques, and similarities and differences found among groups and across studies. DESIGN AND METHODS: A narrative approach was used to review 59 articles published between 1980 and 2000, with particular attention to race, ethnicity, and/or culture. Primary sources of relevant articles included electronic databases, other review articles, and reference lists. RESULTS: Caregiving experiences and outcomes varied across racial and ethnic groups. However, the use of nontheoretical approaches, nonprobability samples, and inconsistent measures among studies has limited understanding of caregiving among diverse populations. IMPLICATIONS: It is important that conceptualization and theorizing broaden with increasing numbers of studies that include diverse groups. Future caregiving research needs to give more attention to such issues as acculturation, assimilation, and cultural values, beliefs, and norms, while also improving theoretical and methodological rigor. PMID- 11914468 TI - Exercise prescribing: computer application in older adults. AB - PURPOSE: The purpose of this study was to determine if older adults are capable and willing to interact with a computerized exercise promotion interface and to determine to what extent they accept computer-generated exercise recommendations. DESIGN AND METHODS: Time and requests for assistance were recorded while 34 college-educated volunteers, equal numbers of men and women, ranging in age from 60 to 87, interfaced with a health promotion tool. The computerized exercise promotion tool's ease of use and the acceptability of the exercise recommendations made were rated by the participants. RESULTS: On average, completion of the items on the computer took 33 min and each participant made 3 requests for assistance, of which only 22% were mouse related. The system's ease of use and the exercise prescription acceptability ratings were high and independent of prior experience with computers. IMPLICATIONS: User friendliness of computerized health promotion tools will determine if, and how, health providers integrate these new technologies into daily practice. The participants in the study were able to complete the computerized items within a reasonable amount of time and with minimal assistance from the provider. These data support the potential of interactive technology in health promotion among the expanding older population. PMID- 11914470 TI - C-cell hyperplasia and medullary thyroid microcarcinoma. AB - Since the discovery of the thyroid C-cell, considerable progress has been made regarding its origin, function, and pathology. In this article an attempt is made to summarize and update our knowledge about physiologic or reactive C-cell hyperplasia, neoplastic C-cell hyperplasia (medullary carcinoma in situ), and medullary microcarcinoma. Seldom recognized preoperatively, physiologic C-cell hyperplasia is associated with inflammatory, metabolic, and neoplastic thyroid disorders as well as with hypercalcemia. However, the pathogenesis is still unclear. Although physiologic C-cell hyperplasia may progress to medullary carcinoma, the full malignant potential is unknown. Problems related to the definition of physiologic C-cell hyperplasia are discussed. Immunohistochemistry and quantitative analysis are required for the diagnosis. By contrast, C-cell hyperplasia associated with MEN II syndromes or familial medullary carcinoma can be diagnosed preoperatively in asymptomatic children or adolescents by the detection of germline mutations of the RET protooncogene. Morphologic and genetic abnormalities support the idea that C-cells in the familial form of C-cell hyperplasia are neoplastic and can be recognized with conventional stains. Therefore, the number of C-cells is irrelevant for the diagnosis. Medullary microcarcinoma is a neoplasm that measures < 1 cm. The sporadic variant is usually an incidental microscopic finding, whereas the familial form can be diagnosed by genetic testing. Its morphologic features and biologic behavior differ from those of larger medullary carcinomas. The frequency of medullary microcarcinoma will probably increase with the use of genetic testing. PMID- 11914471 TI - Classic and recent special stains used in differential diagnosis of endocrine tumors. AB - During the twentieth century, stains for endocrine cells and tumors were developed from empiric cytologic procedures aimed at modern cytochemical methods. Before the 1970s, endocrine stains were mainly based on silver reaction, although other reactions were also proposed; however, the chemical basis of most of these reactions is still unclear. The development of fluorescence procedures for detecting biogenic amines at the cellular level provided information about endocrine cell function of normal cells and related tumors. However, the application of immunocytochemical reactions brought greater and more definitive insights. Several immunocytochemical markers are now available. Some are specific for a definite cell type, while others detect endocrine differentiation in general. Some of these "pan-endocrine" markers are highly specific, and others are highly sensitive but less specific. They all play a definite role in diagnostic pathology. The use of molecular procedures to detect specific mRNA or genetic mutations of diagnostic interest in endocrine pathology should complement immunophenotyping, especially in some problematic fields, such as that of "poorly differentiated" tumors. PMID- 11914472 TI - The differential diagnosis of lesions involving the sella turcica. AB - The sella turcica and the surrounding area contain several different tissues varying in morphology and cytogenesis. Thus, it is not surprising that a large number of diverse lesions may arise in the sellar region. The most frequent abnormalities are the pituitary adenomas, which based on histology, immunocytochemistry, and transmission electron microscopy can be classified into several distinct entities. Pituitary adenomas originate in and consist of adenohypophyseal cells. They are usually slowly growing benign epithelial tumors, which may be associated with increased hormone secretion or may be endocrinologically nonfunctioning. Pituitary carcinomas also arise in adenohypophyseal cells. They are rare and can be diagnosed only when cerebrospinal and/or systemic metastases are documented. To illustrate the diversity, practical importance, and diagnostic difficulties, four cases were selected for presentation: lymphocytic hypophysitis, thyrotroph hyperplasia, growth hormone-producing pituitary adenoma with neuronal transformation, and composite tumor consisting of adenomatous periodic acid Schiff-positive as well as adrenocorticotropic hormone-immunoreactive adenohypophyseal cells and adrenocortical cells. The first two cases are important from a practical point of view because the proper diagnosis can easily be missed, and appropriate interpretation of the findings is essential to prognosis and treatment. The latter two cases are odd, unusual entities; their histogenesis is unresolved. Study of these and many other cases convinced us that careful and detailed morphologic investigation of lesions involving the sella turcica is of fundamental significance. Histology, immunocytochemistry, transmission electron microscopy, and, in some cases, molecular methods are essential to reach a correct diagnosis and to draw conclusions on histogenesis, growth potential, biologic behavior, prognosis, and therapeutic responsiveness. PMID- 11914474 TI - Differential diagnosis of pheochromocytomas and paragangliomas. AB - Paragangliomas are of two types, sympathetic and parasympathetic, depending on the type of paraganglion in which they arise. The term pheochromocytoma is reserved for tumors arising in the adrenal medulla. These tumors are usually fairly easy to diagnose. However, several areas are the subject of debate, including the identification of malignant potential, the diagnosis of medullary hyperplasia, and the recognition of composite tumors. Some histologic features can cause problems in differential diagnosis. Paragangliomas may have spindle cell morphology or contain pigment, requiring distinction from mesenchymal tumors and melanoma, respectively. Extensive degenerative change in pheochromocytomas may mimic adrenal cortical tumors. This short review addresses the diagnosis of pheochromocytomas and paragangliomas and discusses useful approaches in the aforementioned problem areas. PMID- 11914473 TI - Discerning malignancy in resected adrenocortical neoplasms. AB - Differential diagnosis between adenoma and carcinoma in resected human adrenocortical neoplasms may be one of the most problematic and difficult areas of surgical pathology practice. This is especially true in cases of relatively small adrenocortical tumors not associated with obvious signs of malignancy such as necrosis and/or hemorrhage. In addition, the numbers of these small adrenocortical neoplasms are increasing owing to the widespread application of sophisticated computed tomography and/or magnetic resonance imaging scans. No single parameter can be effective in this differential diagnosis of resected adrenocortical tumor. Histopathologic evaluation using a multivariate scoring system is considered most effective in discerning malignancy and biologic behavior of resected adrenocortical neoplasms. Molecular and cellular findings of adrenocortical carcinoma have been inconsistent except for the increased cell proliferation associated with adrenocortical malignancy. Therefore, an assessment of neoplastic cell proliferation using immunostains of cell cycle-associated nuclear antigen such as Ki-67 is the only useful auxiliary method of evaluating malignancy in resected adrenocortical neoplasms at present. PMID- 11914475 TI - Minimally invasive follicular thyroid carcinoma. AB - Infiltration of the capsule, vascular invasion, and/or neoplastic extension into the adjacent parenchyma are regarded as prerequisites for the diagnosis of follicular carcinoma. In modern practice, most of these tumors fall into the category of follicular carcinoma, minimally invasive (FCMI) characterized by evidence of limited capsular or vascular invasion with an excellent long-term prognosis and a good patient outcome. Notwithstanding the wide acceptance of the diagnostic criteria established by the World Health Organization for the classification of follicular carcinomas in particular, they have been difficult to apply and have led to a great deal of confusion. This confusion is compounded when applied to "low-grade" or "minimally invasive" follicular carcinoma because of the poor reproducibility of the classification and the variable results reported in the literature. Our surgical colleagues face a similar lack of a standardized treatment for low-grade follicular carcinomas, which leads to unnecessary surgical treatment. Standardization of histologic criteria is necessary to promote confidence and uniformity in the therapeutic approach of these tumors. We believe that a FCMI is defined as an encapsulated follicular tumor (not papillary), with only small to medium vessel invasion within or immediately adjacent to the tumor capsule and/or up to full-thickness capsular transgression without accompanying extension into the thyroid parenchyma with intervening fibrosis. By using these criteria, patients can be managed with conservative surgical excision to yield an excellent long-term patient outcome. PMID- 11914476 TI - Expression of cellular retinol- and retinoic acid-binding proteins in normal and pathologic human parathyroid glands. AB - We have previously reported data establishing the human parathyroid gland as a target organ for vitamin A. In the present study, we identified Ito-like cells in parathyroid glands, suggesting local stores of vitamin A. Furthermore, we used immunohistochemistry to investigate the expression of the cellular retinol binding protein type 1 and the cellular retinoic acid-binding protein type 1 (CRABP I) in histologically normal glands, in remnants of "normal" glandular tissue adjacent to adenoma, in adenomas, and in hyperplastic glands of chief cell type. All normal and abnormal glands displayed immunoreactivity to the two antibodies. CRABP I appeared in the cytoplasm, cell membranes, and nuclear membranes in normal glands, but only exceptionally in the nuclear membranes in abnormal glands. Since retinoic acid inhibits the secretion of parathyroid hormone and CRABP I is thought to play a key role in regulating the amount of retinoic acid available to interact with specific nuclear receptors, these data may suggest impaired transport of retinoic acid to cell nuclei, thus contributing to the development of hyperparathyroidism. PMID- 11914477 TI - Juvenile rheumatoid arthritis with amyloid goiter: report of a case with review of the literature. AB - Clinically significant enlargement of the thyroid owing to amyloid deposition is a rare occurrence. A 23-yr-old female, a case of juvenile rheumatoid arthritis, developed rapidly increasing thyromegaly during the course of her illness with complaints of dyspnea and dysphagia. Thyroid function tests were within normal limits. Fine-needle aspiration cytology proved inconclusive. Total thyroidectomy was done for symptomatic relief with a preoperative clinical impression of malignancy. Histopathologic findings were consistent with amyloid goiter. The findings of this case are presented, to emphasize the difficulties in making a definite preoperative diagnosis, along with a brief review of the literature. PMID- 11914479 TI - Structure of a Nudix protein from Pyrobaculum aerophilum reveals a dimer with two intersubunit beta-sheets. AB - Nudix proteins, formerly called MutT homolog proteins, are a large family of proteins that play an important role in reducing the accumulation of potentially toxic compounds inside the cell. They hydrolyze a wide variety of substrates that are mainly composed of a nucleoside diphosphate linked to some other moiety X and thus are called Nudix hydrolases. Here, the crystal structure of a Nudix hydrolase from the hyperthermophilic archaeon Pyrobaculum aerophilum is reported. The structure was determined by the single-wavelength anomalous scattering method with data collected at the peak anomalous wavelength of an iridium-derivatized crystal. It reveals an extensive dimer interface, with each subunit contributing two strands to the beta-sheet of the other subunit. Individual subunits consist of a mixed highly twisted and curved beta-sheet of 11 beta-strands and two alpha helices, forming an alpha-beta-alpha sandwich. The conserved Nudix box signature motif, which contains the essential catalytic residues, is located at the first alpha-helix and the beta-strand and loop preceding it. The unusually short connections between secondary-structural elements, together with the dimer form of the structure, are likely to contribute to the thermostability of the P. aerophilum Nudix protein. PMID- 11914478 TI - Histologic, immunohistochemical, and ultrastructural findings in a case of minocycline-associated "black thyroid". AB - We report on a 37-yr-old woman with known antemortem ingestion of minocyclin who died suddenly from a ruptured cerebral aneurysm. At autopsy, her thyroid gland, although not enlarged, was diffusely black, caused by the deposition of a melanin like pigment that stained positive with Schmorl's reagent. The pigment could be bleached with permanganate, and on examination by electron microscopy, it appeared to be deposited within the thyrocyte lysosomes. Additional immunostaining with many antibodies revealed an increase in vimentin staining in the follicular epithelium compared with normal control thyroid glands. Staining for cytoplasmic thyroglobulin was markedly reduced, despite normal thyroid indices performed on stored antemortem blood. Stainable ubiquitin in the follicular epithelium appeared reduced compared with control thyroid tissues. These immunohistochemical findings may reflect disruptions of lysosomal transport and function associated with the abnormal accumulation of pigment. This appears to be the only case of minocyclin-associated "black thyroid" in which extensive immunohistochemical investigations have been performed. PMID- 11914480 TI - Synchrotron white-beam X-ray topography of ribonuclease S crystals. AB - With careful experimental design, indexed synchrotron white-beam X-ray topographs of ribonuclease S crystals at ambient temperature could be recorded with a definition and contrast comparable to that of monochromatic beam topographs of other proteins reported in the literature. By excluding wavelengths longer than 1 A from the white beam with a filter, a radiation dose equivalent to that required to record about 18 topographs could be tolerated without appreciable radiation damage to the samples. Bragg angles of 0.5 degrees or less were required to select low-index harmonically pure reflections with high intensities and extinction lengths only several times the sample's thickness. The resulting X-ray topographs in some cases showed topographic detail and in others showed the even featureless background that has been considered characteristic of a protein crystal of low mosaicity. The ribonuclease S crystals were well ordered single crystals of a quality comparable to other protein crystals that have been studied by X-ray topography. PMID- 11914481 TI - Structure of the photoactive yellow protein reconstituted with caffeic acid at 1.16 A resolution. AB - A structural study is described of the photoactive yellow protein (PYP) reconstituted with the chromophore derivative 3,4-dihydroxycinnamic acid. The crystal structure of PYP reconstituted with this chromophore at 1.16 A resolution is reported in space group P6(5). This is the first high-resolution structure of a photoreceptor containing a modified chromophore. The introduction of an extra hydroxyl group in the native chromophore (i.e. p-coumaric acid) appears to perturb the structure of the hybrid yellow protein only slightly. The chromophore is bound by the protein in two different conformations, separated by a rotation of 180 degrees of the catechol ring. In combination with available spectroscopic data, it is concluded that the caffeic acid chromophore binds to the protein in a strained conformation, which leads to a faster ejection from the chromophore binding pocket upon pB formation. PMID- 11914482 TI - A structural double-mutant cycle: estimating the strength of a buried salt bridge in barnase. AB - Double-mutant cycles are widely used in the field of protein engineering to measure intermolecular and intramolecular interactions. Ideally, there should be no structural rearrangement of the protein on making the two single mutations and the double mutation within the cycle. However, structural pertubation on mutation does not preclude the use of this method, providing the sum of the changes in the single mutants equals the change in the double mutant. In this way, the energy associated with any structural rearrangement cancels in the double-mutant cycle. Previously, the contribution of a buried salt bridge between Arg69 and Asp93 in barnase to the stability of the folded protein has been determined by double mutant cycle analysis. In order to determine whether the measured interaction of 14.0 kJ mol(-1) represents the true interaction energy, the crystal structure of each mutant within the double-mutant cycle was solved. Although mutation results in structural shifts, the majority of those in the single mutants are also found in the double mutant; their energetic effects in the double-mutant cycle are therefore cancelled. This study highlights the robust nature of the double-mutant cycle analysis. PMID- 11914483 TI - Structure of the 1:1 netropsin-decamer d(CCIICICCII)2 complex with a single bound netropsin. AB - The crystal structure of the 1:1 complex of netropsin and the B-DNA decamer d(CCIICICCII)(2) has been elucidated and refined to an R factor of 19.6% and an R(free) of 24.7% using 1790 reflections in the resolution range 8-2.4 A. The complex crystallizes in space group C2, with unit-cell parameters a = 62.40, b = 24.47, c = 36.31 A, beta = 110.09 degrees and one molecule of netropsin in the asymmetric unit; the rest of the minor groove is filled with six water molecules. The structure was solved by the molecular-replacement method using the DNA model d(CCCCCIIIII)(2) from the 2:1 netropsin complex by removing both bound netropsins (Chen et al., 1998). Surprisingly, only one netropsin molecule is found to bind to the present decamer, covering residues 2-6 at the upper stream of the duplex. The positively charged guanidinium head is hydrogen bonded through N1H(2) to the O(2) of cytosine 2 and through N10H(2) to N(3) of inosine 6. The three amide N-H groups of the peptides face the minor groove and form three sets of bifurcated hydrogen bonds with the base atoms. The central part of the drug (C3-N8) is nearly conjugated. The preference of the cytosine carbonyl O2 atoms over the inosine N3 atoms in hydrogen bonding is seen. The drug-bound region has more uniform twists, roll angles, propeller twists and minor-groove widths compared with the water-bound region. PMID- 11914485 TI - Specificity of TRAP-RNA interactions: crystal structures of two complexes with different RNA sequences. AB - The trp RNA-binding attenuation protein (TRAP) regulates expression of the tryptophan biosynthetic genes in bacilli by binding to the leader region of the nascent trp operon mRNA. When activated by binding tryptophan, the 11-subunit circular TRAP molecule binds to a target sequence consisting of 11 (G/U)AG repeats, separated by two or three variable 'spacer' nucleotides. Reported here are two crystal structures of TRAP bound to RNAs containing 11 GAG repeats separated by UU and CC spacer nucleotides, determined at 1.75 and 2.50 A resolution, respectively. These show the spacer regions of the RNA molecules to be highly flexible, making no direct hydrogen-bonding contacts with the protein. Comparison of these structures with the previous structure of TRAP bound to (GAGAU)(10)GAG RNA, in which the spacer nucleotides stack with each other close to the protein surface, shows that the RNA can adopt different conformations depending on the sequence of the spacer regions. This gives insight into the structural basis of the specificity of TRAP and into the mechanism of binding. PMID- 11914484 TI - Cryophotolysis of caged compounds: a technique for trapping intermediate states in protein crystals. AB - Caged compounds in combination with protein crystallography represent a valuable tool in studies of enzyme reaction intermediates. To date, photochemical triggering of reactions has been performed close to room temperature. Synchronous reaction initiation has only been achieved with enzymes of relatively slow turnover (<0.1 s(-1)) and caged compounds of high quantum yield. Here X-ray crystallography and microspectrophotometry were used to provide evidence that (nitrophenyl)ethyl (NPE) ester bonds can be photolyzed by UV light at cryotemperatures. NPE-caged ATP in flash-cooled crystals of Mycobacterium tuberculosis thymidylate kinase was photolyzed successfully at 100-150 K as assessed by the structural observation of ATP-dependent enzymatic conversion of TMP to TDP after temporarily warming the crystals to room temperature. A new method is proposed in which cryo-photolysis combined with temperature-controlled protein crystallography can be used to trap reaction intermediates even in some of the fastest enzymes and/or when only compounds of low quantum yield are available. Raising the temperature after cryophotolysis may allow a transition barrier to be passed and an intermediate to accumulate in the crystal. A comparable method has only been used so far with proteins displaying endogenous photosensitivity. The approach described here opens the way to studying the reaction mechanisms of a much larger number of crystalline enzymes. Furthermore, it is shown that X-ray-induced radiolysis of caged compounds occurs if high intensity synchrotron beamlines are used. This caveat should be taken into account when deriving data-collection protocols. It could also be used potentially as a way to trigger reactions. PMID- 11914486 TI - The structure of human recombinant aldose reductase complexed with the potent inhibitor zenarestat. AB - The crystal structure of the complex of human recombinant aldose reductase (AR) with zenarestat, one of its potent inhibitors, has been solved at 2.5 A resolution. Zenarestat fits neatly in the hydrophobic active site and induces unique and dramatic conformational changes. For example, the benzene ring of zenarestat occupies a gap in the side chains of Leu300 and Trp111 that interact directly and forms a CH-pi interaction in the native holoenzyme. As a result, the benzene ring of the inhibitor and these side chains form a CH-pi-pi interaction. Such structural information is key to understanding the mode of action of this class of inhibitors and for rational design of better therapeutics. PMID- 11914487 TI - Structure of an RNase-related protein from Calystegia sepium. AB - The structure of a catalytically inactive RNase-related protein from Calystegia sepium (CalsepRRP) has been resolved by protein crystallography at a resolution of 2.05 A and an R factor of 20.74%. Although the protein is completely devoid of ribonuclease activity, it adopts the typical alpha + beta structure of non-base specific RNases. Analysis of the structure revealed that two amino-acid substitutions in the 'active' P1 site, in combination with the less hydrophobic/aromatic character of the B1 base-recognition site and a completely disrupted B2 base-recognition site, might account for this complete lack of activity. PMID- 11914488 TI - X-ray crystallographic analyses of symmetrical allosteric effectors of hemoglobin: compounds designed to link primary and secondary binding sites. AB - The rational design and X-ray crystallographic analyses of two symmetrical allosteric effectors of hemoglobin (Hb) are reported. Compound design was directed by the previously solved co-crystal structure of one of the most potent allosteric effectors of Hb, 2-[4-[(3,5-dichlorophenylcarbamoyl)-methyl]-phenoxy] 2-methylpropionic acid (RSR4), which revealed two distinct binding sites for this compound in the Hb central water cavity. The primary binding site has been observed for all compounds of this structural class, which stabilize deoxy Hb by engaging in inter-dimer contacts with three of the four protein subunits. Interactions at the secondary binding site of RSR4 occur primarily between the beta(1) and beta(2) subunits and serve to further constrain the deoxy state. Based on these observations, it was hypothesized that compounds with the ability to simultaneously span and link both of these sites would possess increased potency, but at a lower molar concentration than RSR4. Two symmetrical compounds were designed and synthesized based on this hypothesis. The symmetrical effector approach was taken to minimize the number of compound orientations needed to successfully bind at either of the distinct allosteric sites. X-ray crystallographic analyses of these two effectors in complex with Hb revealed that they successfully spanned the RSR4 primary and secondary binding sites. However, the designed compounds interacted with the secondary binding site in such a way that intra-dimer, as opposed to inter-dimer, interactions were generated. In agreement with these observations, in vitro evaluation of the symmetrical effectors in Hb solution indicated that neither compound possessed the potency of RSR4. A detailed analysis of symmetrical effector-Hb contacts and comparisons with the binding contacts of RSR4 are discussed. PMID- 11914489 TI - Comparative analysis of space-grown and earth-grown crystals of an aminoacyl-tRNA synthetase: space-grown crystals are more useful for structural determination. AB - Protein crystallization under microgravity aims at benefiting from the quasi absence of convection and sedimentation to favor well ordered crystal nucleation and growth. The dimeric multidomain enzyme aspartyl-tRNA synthetase from Thermus thermophilus has been crystallized within dialysis reactors of the Advanced Protein Crystallization Facility in the laboratory on earth and under microgravity aboard the US Space Shuttle. A strictly comparative crystallographic analysis reveals that the crystals grown in space are superior in every respect to control crystals prepared in otherwise identical conditions on earth. They diffract X-rays more intensely and have a lower mosaicity, facilitating the process of protein structure determination. Indeed, the electron-density map calculated from diffraction data of space-grown crystals contains considerably more detail. The resulting three-dimensional structure model at 2.0 A resolution is more accurate than that produced in parallel using the data originating from earth-grown crystals. The major differences between the structures, including the better defined amino-acid side chains and the higher order of bound water molecules, are emphasized. PMID- 11914490 TI - Structure of an endoglucanase from termite, Nasutitermes takasagoensis. AB - Contrary to conventional wisdom, it has been shown recently that termites do not necessarily depend on symbiotic bacteria to process cellulose. They secrete their own cellulases, mainly endo-beta-1,4-glucanase and beta-1,4-glucosidase. Here, the first structure of an endogenous endoglucanase from the higher termite Nasutitermes takasagoensis (NtEgl) is reported at 1.40 A resolution. NtEgl has the general folding of an (alpha/alpha)(6) barrel, which is a common folding pattern for glycosyl hydrolase family 9. Three-dimensional structural analysis shows that the conserved Glu412 is the catalytic acid/base residue and the conserved Asp54 or Asp57 is the base. The enzyme has a Ca(2+)-binding site near its substrate-binding cleft. Comparison between the structure of the Ca(2+)-free enzyme produced by reducing the pH of the soaked crystal from 5.6 (the pH of optimum enzyme activity) to 2.5 with that of the Ca(2+)-bound enzyme did not show significant differences in the locations of the C(alpha) atoms. The main differences are in the conformation of the residue side chains ligating the Ca(2+) ion. The overall structure of NtEgl at pH 6.5 is similar to that at pH 5.6. The major change observed was in the conformation of the side chain of the catalytic acid/base Glu412, which rotates from a hydrophobic cavity to a relatively hydrophilic environment. This side-chain displacement may decrease the enzyme activity at higher pH. PMID- 11914491 TI - Determination of the structure of an endoglucanase from Aspergillus niger and its mode of inhibition by palladium chloride. AB - The fungus Aspergillus niger is a main source of industrial cellulase. beta-1,4 Endoglucanase is the major component of cellulase from A. niger. In spite of widespread applications, little is known about the structure of this enzyme. Here, the structure of beta-1,4-endoglucanase from A. niger (EglA) was determined at 2.1 A resolution. Although there is a low sequence identity between EglA and CelB2, another member of family 12, the three-dimensional structures of their core regions are quite similar. The structural differences are mostly found in the loop regions, where CelB2 has an extra beta-sheet (beta-sheet C) at the non reducing end of the binding cleft of the native enzyme. Incubation of EglA with PdCl(2) irreversibly inhibits the EglA activity. Structural studies of the enzyme palladium complex show that three Pd(2+) ions bind to each EglA molecule. One of the Pd(2+) ions forms a coordinate covalent bond with Met118 S(delta) and the nucleophilic Glu116 O(epsilon1) at the active site of the enzyme. The other two Pd(2+) ions bind on the surface of the protein. Binding of Pd(2+) ions to EglA does not change the general conformation of the backbone of the protein significantly. Based on this structural study, one can conclude that the palladium ion directly binds to and blocks the active site of EglA and thus inactivates the enzyme. PMID- 11914492 TI - Crystallization and preliminary X-ray study of an N-terminal fragment of rat liver ribosomal P2 protein. AB - Ribosomal P proteins have been shown to be involved in the binding of elongation factors and participate in factor-dependent GTP hydrolysis. The P proteins form the pentamer (P1/P2)(2)-P0 constituting the lateral flexible stalk of the 60S ribosomal subunit. The highly soluble domain (1-65) of rat liver P2 has been overexpressed in Escherichia coli as an N-terminal poly-His-tagged protein and crystallized. To reduce nucleation and improve crystal morphology and diffraction power, the crystals were grown in a gel matrix and an oil barrier was added between the reservoir and the drop to reduce the rate of vapour diffusion. This dramatically reduced the nucleation in the drops and yielded diffraction-quality crystals. Data were collected to 2.4 A resolution at beamline ID 14-1, ESRF. The crystals belong to the orthorhombic space group P2(1)2(1)2, with unit-cell parameters a = 37.7, b = 96.7, c = 135.0 A. PMID- 11914493 TI - Crystallization and preliminary X-ray analysis of the complex of porcine pancreatic elastase and a hybrid squash inhibitor. AB - A hybrid inhibitor consisting of the scaffold of a squash-type inhibitor and a specific inhibitory peptide optimized from the third domain of ovomucoid inhibitor from turkey against porcine pancreatic elastase was synthesized by peptide synthesis. The complex formed by this hybrid inhibitor and the porcine pancreatic elastase was crystallized using the hanging-drop method with citrate in the crystallization solution. The space group was determined to be P2(1)2(1)2(1), with unit-cell parameters a = 56.33, b = 56.44, c = 72.76 A. A complete X-ray diffraction data set was collected under cryogenic conditions to 1.8 A. PMID- 11914494 TI - Purification, crystallization and preliminary X-ray crystallographic analysis of agkaggregin, a C-type lectin-like protein from Agkistrodon acutus venom. AB - A snake-venom C-type lectin-like protein, agkaggregin, has been isolated from Agkistrodon acutus venom. Agkaggregin has an apparent molecular mass of about 28 kDa and consists of two different types of subunits, an alpha-subunit (approximately 15 kDa) and a beta-subunit (approximately 14 kDa). Agkaggregin has the ability to induce platelet aggregation at concentrations of the order of nanomoles. The agkaggregin crystals grew for nearly a year by hanging-drop vapour diffusion and belong to the I222 space group, with unit-cell parameters a = 64.75, b = 74.21, c = 133.24 A. One asymmetric unit contains one alphabeta heterodimer, corresponding to a volume-to-mass ratio of 2.795 A(3) Da(-1). Agkaggregin may exist in two association forms: an alphabeta heterodimer and a dimer of alphabeta heterodimers that associates during the long process of crystallization. PMID- 11914495 TI - Crystallization and preliminary X-ray crystallographic studies of a complex between the Lactococcus lactis Fpg DNA-repair enzyme and an abasic site containing DNA. AB - For protein-DNA complex crystallization, the choice of the DNA fragment is crucial. With the aim of crystallizing the 31 kDa Fpg DNA-repair enzyme bound to DNA, oligonucleotide duplexes varying in length, sequence, end type and nature of the specific DNA target site were used. Crystals of several protein-DNA combinations grew from solutions containing both polyethylene glycol and salt. This systematic crystallization screening followed by optimization of the crystallization conditions by microseeding led to crystals of Fpg bound to a 13 base-pair duplex DNA carrying the 1,3-propanediol abasic site analogue which are suitable for crystallographic analysis. Complete native data sets have been collected to 2.1 A resolution. PMID- 11914496 TI - Expression, purification and preliminary crystallographic studies of alpha amylase isozyme 1 from barley seeds. AB - The germinating barley seed contains two major alpha-amylase isozyme families, AMY1 and AMY2, involved in starch degradation to provide energy used by the plant embryo for growth. Many years of difficulty in growing three-dimensional crystals of natural AMY1 have now been overcome by a nonapeptide truncation of the enzyme C-terminus. The truncated enzyme was overexpressed in Pichia pastoris, purified and crystallized by the hanging-drop vapour-diffusion method using polyethylene glycol 8000 as precipitant and 2-propanol as an additive. Crystals belong to the orthorhombic space group P2(1)2(1)2, with unit-cell parameters a = 88.36, b = 72.82, c = 61.74 A and one molecule per asymmetric unit. PMID- 11914497 TI - Crystallization and preliminary X-ray diffraction analysis of fatty-acid hydroxylase cytochrome P450BSbeta from Bacillus subtilis. AB - Cytochrome P450 isolated from Bacillus subtilis (P450BSbeta; MW 48 kDa) catalyzes the hydroxylation of long-chain fatty acids at the alpha and beta positions using H(2)O(2) as an oxidant. Crystals of the substrate-free form of P450BSbeta belonging to the trigonal space group P3(2)21 or P3(1)21 were obtained by the sitting-drop vapour-diffusion method using a precipitate solution consisting of 10%(w/v) PEG 4000 and 50 mM MES pH 6.8. Another crystal form, belonging to the rhombohedral space group R3 or R32, was obtained from precipitate solution consisting of 10% PEG 4000, 0.15 mM magnesium acetate and 50 mM MES pH 6.5 in the presence of 2 mM myristic acid (substrate). Using synchrotron radiation, both P450BSbeta crystals diffracted to 2.5 A resolution. Bijvoet and dispersive anomalous difference Patterson maps show a clear peak corresponding to the haem iron. PMID- 11914498 TI - Crystallization and characterization of the dehydrogenase domain from rat peroxisomal multifunctional enzyme type 1. AB - Peroxisomal multifunctional enzyme type 1 from rat (perMFE-1) is a monomeric multidomain protein shown to have 2-enoyl-CoA hydratase/Delta(3)-Delta(2)-enoyl CoA isomerase and (3S)-hydroxyacyl-CoA dehydrogenase domains followed by a C terminal extension of 130 amino acids with unknown function apart from being a carrier of the peroxisomal targeting signal type 1. The truncated perMFE-1 without the N-terminal hydratase/isomerase domain (perMFE-1DH; residues 260-722) was overexpressed as an enzymatically active recombinant protein, purified and characterized. Using (3S)-hydroxydecanoyl-CoA as a substrate, the specific enzymatic activity of perMFE-1DH was determined to be 2.2 micromol min(-1) mg( 1), comparable with that of perMFE-1 purified from rat liver (2.8 micromol min( 1) mg(-1)). The protein was crystallized in the apo form by the hanging-drop method and a complete data set to 2.45 A resolution was collected using a rotating-anode X-ray source. The crystals have primitive tetragonal symmetry, with unit-cell parameters a = b = 125.9, c = 60.2 A. PMID- 11914499 TI - Purification, crystallization and preliminary X-ray diffraction studies of a Ca2+ binding protein, human S100P. AB - S100P, a Ca(2+)-binding protein, is a member of the S100 family. Its presence is associated with the development of prostate cancer, but its cellular function is not known. Recombinant human S100P has been expressed and purified in bacterial cells and crystals of human S100P in the calcium-bound state have been grown using the vapour-diffusion technique with PEG 4000 as precipitant. Diffraction data have been obtained to a resolution of 2.0 A from a single frozen S100P crystal which belongs to the space group P4(1)2(1)2, with unit-cell parameters a = b = 60.8, c = 47.6 A. PMID- 11914500 TI - Crystallization and preliminary X-ray diffraction analysis of two pH-dependent forms of a di-haem cytochrome c peroxidase from Pseudomonas nautica. AB - Two crystal forms of cytochrome c peroxidase from Pseudomonas nautica were obtained, one at pH 4.0 using sodium citrate as precipitant and another at pH 5.3 using ammonium phosphate and sodium citrate as precipitants. The two forms belong to different space groups P3(1)21 (pH 4.0) and P6(4)22 (pH 5.3), with unit-cell parameters a = b = 114.5, c = 90.7 A and a = b = 151.0, c = 155.9 A, respectively. Several complete data sets were collected using synchrotron radiation at ESRF and Cu K(alpha) X-ray radiation from a rotating-anode generator. These results will contribute to clarifying the haem transitions occurring during peroxidatic reaction and the required electron-transfer processes and to elucidating the catalytic mechanism of the enzyme and the role of calcium in the activation process. PMID- 11914501 TI - Protein engineering for crystallization of the GTPase Sar1 that regulates ER vesicle budding. AB - Sar1 is an important and unique GTPase that regulates vesicle budding from the ER membrane. An effort to crystallize full-length hamster Sar1 was unsuccessful owing to the aggregation of Sar1 in solution as indicated by dynamic light scattering measurements. It was presumed that a patch of hydrophobic residues in the N-terminal region of Sar1 was responsible for the aggregation. To attempt to improve protein crystallizability, the N-terminal residues of Sar1 were progressively truncated and the solution behavior of the resulting Sar1 variants was monitored by dynamic light scattering. Truncation of the first nine residues from the N-terminus led to a Sar1 variant that is monodisperse in solution. This well behaved Sar1 variant yielded crystals in just a few days that were ultimately refined to diffraction quality. PMID- 11914502 TI - Crystallization and preliminary crystallographic analysis of a novel cytochrome P450 from Mycobacterium tuberculosis. AB - The product of the Rv2276 gene of Mycobacterium tuberculosis is a cytochrome P450 (P450 MT2, CYP121) which has been shown to bind tightly to a range of azole-based antifungal drugs (e.g. miconazole, clotrimazole). These drugs are potent inhibitors of mycobacterial growth, suggesting that P450 MT2 (CYP121) may be a potential drug target. The enzyme has been overexpressed in Escherichia coli and crystallized by the hanging-drop method. Crystals of P450 MT2 (CYP121) belong to the hexagonal space group P6(1)22 or P6(5)22, with unit-cell parameters a = b = 78.3, c = 265.6 A. Native data have been collected to 1.6 A resolution and Hg derivative data to 2.5 A resolution using a synchrotron-radiation source. PMID- 11914503 TI - Crystallization and preliminary X-ray analysis of the selenate reductase from Thauera selenatis. AB - Selenate reductase from Thauera selenatis was crystallized using ammonium sulfate as a precipitant. Crystals of selenate reductase belong to the space group C2, with unit-cell parameters a = 116.9, b = 67.5, c = 186.7 A, beta = 90 degrees. Native data to 2.1 A resolution have been collected and a heavy-atom derivative has been identified following soaking of the crystals in a solution of trimethyl lead acetate. PMID- 11914504 TI - Crystallization and preliminary X-ray characterization of a thermostable pectate lyase from Thermotoga maritima. AB - Pectate lyase is an enzyme involved in the degradation of the pectate portion of the primary plant cell wall. A recombinant pectate lyase from Thermotoga maritima where three of the four cysteine residues have been mutated (C132I, C156N, C194L) has been crystallized. Crystals of the same morphology and trigonal space group R3 with similar unit-cell parameters were obtained under two different conditions. The first, 0.3 M (NH(4))H(2)PO(4) pH 4.2, gave crystals with a maximum size of 0.4 x 0.2 x 0.2 mm in one week that diffracted to a resolution of 1.87 A and had unit-cell parameters a = b = 80.6, c = 148.8 A. The second, 0.1 M sodium acetate, 6%(w/v) PEG 4000 pH 6.5, gave the same size crystals in two weeks that diffracted to a resolution of 2.1 A and had unit-cell parameters a = b = 80.0, c = 150.1 A. PMID- 11914505 TI - Purification and crystallization of the yeast elongation factor eEF2. AB - Crystals of the Saccharomyces cerevisiae elongation factor 2 (eEF2) in complex with GDP were obtained with the vapour-diffusion technique after rapid purification from industrial yeast. The crystals diffract to 2.85 A and belong to the space group P2(1)2(1)2(1). A yeast strain expressing a functional histidine tagged eEF2 as the only form of the protein further allows facilitated purification of the factor for both structural and functional studies. PMID- 11914506 TI - Purification, crystallization and preliminary X-ray diffraction of Cys103Ala acyl coenzyme A: isopenicillin N acyltransferase from Penicillium chrysogenum. AB - Penicillins and cephalosporins are an efficacious group of antibiotics produced by fungi such as Penicillium chrysogenum and Acremonium chrysogenum. The last step in their biosynthesis is catalyzed by acyl coenzyme A:isopenicillin N transferase (AT). This enzyme is produced as a single-chain proenzyme, which is activated by autocatalytic cleavage of the Gly102-Cys103 peptide bond, resulting in a heterodimeric protein with subunits of 11 and 29 kDa. The Cys103Ala mutant of the proenzyme, which does not undergo this cleavage, was purified and crystallized. Diffraction-quality crystals of the mutant and an L-SeMet substituted mutant were obtained by vapour diffusion against solutions containing (NH(4))(2)SO(4), NaCl and HEPES-NaOH pH 7.5. The crystals belong to the monoclinic space group C2, with unit-cell parameters a = 231.36, b = 68.27, c = 151.31 A and beta = 129.56 degrees. They diffract to 2.8 A resolution with X-rays from a rotating-anode generator. PMID- 11914507 TI - Structural plasticity in the eight-helix fold of a trematode haemoglobin. AB - The three-dimensional structure of recombinant haemoglobin from the trematode Paramphistomum epiclitum, displaying the highest oxygen affinity so far observed for (non)vertebrate haemoglobins, has previously been determined at 1.17 A resolution (orthorhombic space group P2(1)2(1)2(1)). In the present communication, the three-dimensional structure of wild-type P. epiclitum haemoglobin is reported at 1.85 A resolution in a monoclinic crystal form (R factor = 16.1%, R(free) = 22.0%). Comparison of P. epiclitum (recombinant versus wild-type ferric Hb) structures in the two crystal forms shows structural differences in the haem proximal and distal sites which have not been reported for other known haemoglobin structures previously. PMID- 11914508 TI - Crystallization and preliminary crystallographic studies of Streptococcus pyogenes cysteine protease precursor. AB - Streptococcal protease precursor, secreted by the human pathogen Streptococcus pyogenes, becomes activated to a cysteine protease. The precursor and the mature enzyme appear to contribute to S. pyogenes virulence. The precursor protein was crystallized in the form of very thin flexible flakes. X-ray diffraction data were collected to 3.15 A resolution at 100 K using synchrotron radiation. The crystals are monoclinic, space group P2(1), with unit-cell parameters a = 41.6, b = 136.0, c = 156.7 A, beta = 95.7 degrees, and contain four copies of the protein in the asymmetric unit. PMID- 11914509 TI - Linear regression calibration: theoretical framework and empirical results in EPIC, Germany. AB - Large scale dietary assessment instruments are usually based on the food frequency technique and have therefore to be tailored to the involved populations with respect to mode of application and inquired food items. In multicenter studies with different populations, the direct comparability of dietary data is therefore a challenge because each local dietary assessment tool might have its specific measurement error. Thus, for risk analysis the direct use of dietary measurements across centers requires a common reference. For example, in the European prospective cohort study EPIC (European Prospective Investigation into Cancer and Nutrition) a 24-hour recall was chosen to serve as such a reference instrument which was based on a highly standardized computer-assisted interview (EPIC-SOFT). The 24-hour recall was applied to a representative subset of EPIC participants in all centers. The theoretical framework of combining multicenter dietary information was previously published in several papers and is called linear regression calibration. It is based on a linear regression of the food frequency questionnaire to the reference. The regression coefficients describe the absolute and proportional scaling bias of the questionnaire with the 24-hour recall taken as reference. This article describes the statistical basis of the calibration approach and presents first empirical results of its application to fruit, cereals and meat consumption in EPIC Germany represented by the two EPIC centers, Heidelberg and Potsdam. It was found that fruit could be measured well by the questionnaire in both centers (lambdacirc; = 0.98 (males) and lambdacirc; = 0.95 (females) in Heidelberg, and lambdacirc; = 0.86 (males) and lambdacirc; = 0.7 (females) in Potsdam), cereals less (lambdacirc; = 0.53 (males) and lambdacirc; = 0.4 (females) in Heidelberg, and lambdacirc; = 0.53 (males) and lambdacirc; = 0.44 (females) in Potsdam), and that the assessment of meat (lambdacirc; = 0.72 (males) and lambdacirc; = 0.65 (females) in Heidelberg, and lambdacirc; = 0.49 (males) and lambdacirc; = 0.42 (females) in Potsdam) has a center-specific bias. The application of the calibration approach to the questionnaire data will change the ranking of the two centers following the data of the reference instrument, and not well-measured food items will exhibit considerably less variation compared to the original data. We conclude that calibration is a necessary step in multicenter studies. However, this exercise shows that the current statistical framework is not yet sufficiently developed for a broad application. PMID- 11914510 TI - Effect of heat-treated proteins on selected parameters of the biotransformation system in the rat. AB - The intake of heat-damaged proteins from food causes various effects, like the loss of essential amino acids and a reduced protein digestibility. There is also an influence on gastrointestinal microorganisms and different digestion enzymes. Until now, very little is known about the influence of heat-treated proteins on the enzymes of the biotransformation system. In the present study, the influence of protein-bound L-lysino-D,L-alanine, N(epsilon)-fructoselysine, and N(epsilon) carboxymethyllysine (CML) on selected enzymes of the biotransformation in liver, kidney, and intestinal mucosa of male Wistar rats was examined. The contents of cytochrome P-450 and cytochrome b(5) and the activity of NADPH-cytochrome c reductase served as indicators of phase I biotransformation. The influence on phase II biotransformation was shown by the content of glutathione and the glutathione S-transferase activity. The results showed that treatment with heat damaged proteins mainly affected phase II biotransformation enzymes with CML, yielding the strongest effect. The activity of glutathione S-transferase in the kidney was 86% higher in animals treated with diets containing 4,930 mg.kg(-1) protein-bound CML than in animals of the control group which received a diet without any detectable CML. In addition, a higher level of glutathione was found in the kidneys of animals fed on diets containing CML. The glutathione S transferase activity was 64% higher in the intestinal mucosa of animals fed on protein-bound N(epsilon)-fructoselysine (2,700 mg.kg(-1)). The glutathione S transferase activity was higher (p >0.05) in the intestinal mucosa of animals fed on protein-bound L-lysino-D,L-alanine (2,582 and 12,474 mg.kg(-1)). In conclusion, ingestion of heat-treated proteins led to an activation of the enzymes of phase II biotransformation. Whether or not the released pure compounds or the degradation products of the test proteins are responsible for the altered enzyme activities remains to be evaluated. PMID- 11914511 TI - Effect of either gamma-tocotrienol or a tocotrienol mixture on the plasma lipid profile in hamsters. AB - BACKGROUND/AIMS: Tocotrienols has been shown to inhibit the 3-hydroxy-3 methylglutaryl-coenzyme A (HMG-CoA) reductase activity; however, the published animal and human studies yield conflicting results. We investigated the effects of a 4-week dietary supplement of either gamma-tocotrienol (86% gamma-T3) or a mixture of tocotrienols (29.5% alpha-T3, 3.3% beta-T3, 41.4% gamma-T3, 0.1% delta T3: mix-T3) on the plasma lipid profile in hamsters receiving a high fat diet. METHODS: The hamsters were randomized into 7 groups: no treatment, 16 mg/day/kg BW simvastatin, 23, 58, 263 mg/day/kg BW gamma-tocotrienol, and 39 or 263 mg/day/kg BW for the mixture of tocotrienols. Plasma lipid levels were measured after 2 and 4 weeks of treatment. RESULTS: In all groups treated with tocotrienol total cholesterol levels were decreased, ranging from 7 to 23% after 2 weeks of treatment and from 7 to 15% after 4 weeks. Low-density lipoprotein plasma levels changed accordingly: a decline of 6-37% after 2 weeks and of 12-32% at the end of the study was observed. After 4 weeks of treatment, total cholesterol and low density lipoprotein plasma levels were significantly reduced in the 263 mg/day/kg BW mixed tocotrienols and the 58 mg/day/kg BW and 263 mg/day/kg BW gamma tocotrienol groups when compared to the no treatment group. Plasma triglycerides and high-density lipoprotein levels did not change significantly. CONCLUSION: This study provides further evidence that tocotrienols lower total cholesterol and low density lipoprotein plasma levels in hamsters and that gamma-tocotrienol is a more potent agent than a mixture of tocotrienols. PMID- 11914512 TI - Responses to dietary macronutrient distribution of overweight rats under restricted feeding. AB - Changes in protein and amino acid balance after energy-restricted feeding have scarcely been studied, although it has been suggested that protein utilization may depend on the macronutrient composition of the restricted diet. The aim of this study is to investigate the influence of dietary fat quality and content, during an energy-restricted regime, on body composition and protein metabolism. Therefore, diet-induced overweight rats were divided into three dietary groups: one group was fed on a control diet ad libitum (control) and the other two groups were fed energy-restricted diets (about 60% of control group), which provided a standard amount (20%) of fat (SFR), based on olive oil, or a high amount (60%) of saturated fat (HFR), based on coconut oil. Measurements of body weight, body composition, serum biochemical parameters and the assessment of the hepatic and muscular protein response were performed. Similar results were found comparing weight losses and serum parameters in both deprived groups, although the high-fat restricted rats (HFR group) showed a greater reduction in the subcutaneous fat depot and of total body fat. After both energy-restricted treatments, the serum amino acid concentration was reduced while the urinary amino acid excretion increased. Muscle and liver protein metabolism was affected by energy restriction, which produced a decrease in protein synthesis capacity (RNA content) in both tissues and a higher muscle proteolysis (catepsin activity), more marked in the SFR group, while no changes were found in liver protein breakdown. Hepatic glycogen and glycogenic amino acid were also altered, reaching significant differences in the HFR group. Thus, dietary macronutrient composition during energy restriction seems to be involved in the metabolic adaptative response. PMID- 11914513 TI - Postprandial endothelial impairment and reduced glutathione levels in postmenopausal women. AB - BACKGROUND/AIM: Postmenopausal age is characterized by a higher risk for coronary heart disease (CHD) and postprandial lipemia is strictly related with the evidence of CHD. The aim of the study was to clarify the vascular effects of postprandial state in postmenopausal women. METHODS: Ten postmenopausal women (mean age 57 +/- 8 years) without vascular risk factors and history of cardiovascular disease underwent an oral fat load test. Endothelial function, expressed as brachial flow-mediated vasodilation (FMV), lipid parameters and reduced glutathione (GSH) were evaluated at baseline and 2, 4 and 6 h after the load. RESULTS: FMV showed a significant decrease at the 2nd hour (2.3 +/- 2.6%, vs. baseline 7.7 +/- 2.8%, p < 0.05) and overlapping to the basal value after 4 h. Triglycerides increased postprandially at the 2nd and 4th hour (1.6 +/- 0.6 micromol/l, 1.8 +/- 0.5 micromol/l vs. baseline 0.9 +/- 0.4 micromol/l, p < 0.05), decreasing thereafter. GSH decreased at the 2nd hour of the postprandial phase (5.1 +/- 1.9 micromol/l vs. baseline 8.4 +/- 1.9 micromol/l, p < 0.05), normalizing successively. At the univariate analysis a negative correlation was found between FMV and triglyceride changes (r = -0.37, p < 0.05) and a positive one between FMV and GSH modifications (r = 0.40, p < 0.05). CONCLUSION: These data demonstrated that postprandial lipemia transiently impairs endothelial reactivity by an oxidative burden, partly dependent to triglyceride increase. PMID- 11914514 TI - Chronic ethanol feeding affects intestinal mucus lipid composition and glycosylation in rats. AB - Effect of chronic ethanol feeding on mucus lipid composition and glycosylation was investigated in rat intestine. Ethanol feeding to rats daily for 40 days enhanced cholesterol, phospholipids, triglyceride and cerebroside contents of intestinal mucus. Ethanol feeding exhibited a differential effect on various sugar contents of the mucus. Total hexose content was increased (42%) whereas hexosamines, sialic acid and fucose contents were reduced (p < 0.001). Results on incorporation of labeled sugars into mucus revealed an enhanced (p < 0.01) galactosylation and reduced (p < 0.01) glucosamination of glycoproteins. Separation of glycoproteins on SDS-PAGE showed that proteins of molecular weight >200, 118, 65 and 55 kD were affected. These were highly galactosylated and less glucosaminated in ethanol-fed animals compared to controls. Thus, ethanol feeding markedly alters the lipid composition and glycosylation process of intestinal mucus in rats. These alterations may be of pathological significance in view of the role of mucus in protection and adhesion of microorganisms. PMID- 11914515 TI - Myocardial and hepatic free carnitine concentrations in pups of diabetic female rats. AB - BACKGROUND/AIMS: Adequate carnitine levels are required for normal fatty acid and energy metabolism in heart muscle. It is well known that streptozotocin-induced diabetic rats develop myocardial carnitine deficiency and that carnitine therapy may be beneficial to the diabetic heart. Infants of diabetic mothers (IDM) are known to be at risk for developing a hypertrophic type of cardiomyopathy. In the present investigation, we examined the free carnitine concentration from cardiac and hepatic tissue in pups of streptozotocin-induced diabetic female rats. We also assessed the effect of maternal L-carnitine supplementation on the free carnitine concentration in pups of diabetic rats. METHOD: Three groups, each consisting of 4 Wistar albino female rats, were studied, Group 1 (untreated diabetic; n = 4) and group 2 (L-carnitine-treated diabetic; n = 4) rats were given streptozotocin (60 mg/kg) by intraperitoneal injection; group 3 were controls. During pregnancy, L-carnitine was given at a dose of 150 mg/kg by intraperitoneal injection once a day for 14 days. Cesarean section was carried out, and 113 newborn rats (group 1 n = 36; group 2 n = 38; group 3 n = 39) were obtained from all the pregnant rats. RESULTS: The free carnitine concentration in myocardial tissue was significantly decreased in the female diabetic rats (p < 0.001). However, the free carnitine concentration from hepatic tissue in diabetic female rats was similar to that in controls. In pups of group 1 diabetic rats, a significantly decreased free carnitine concentration was found in both myocardial and hepatic tissue compared to group 2 and controls. The free carnitine content from myocardial and hepatic tissue was significantly elevated in the maternal L carnitine-supplemented group when compared to group 1 and control pups (p < 0.001 and p < 0.05, respectively). CONCLUSIONS: The present study has demonstrated that the free carnitine concentration from myocardial and hepatic tissue is significantly reduced in pups of streptozotocin-induced diabetic female rats. This study has also shown that administration of maternal L-carnitine improves the carnitine level in pups of diabetic rats. A decreased myocardial carnitine concentration may be partly responsible for the development of cardiomyopathy in IDM. PMID- 11914517 TI - Cochlear transduction: from models to molecules and back again. AB - The strides made over the last few years towards understanding many details of cochlear function still leave a number of issues unresolved. Integrating the information from molecular, genetic and, increasingly, genomic sources requires models that provide close matching between data and theory. For both theoretical and experimental reasons, the difficult area in cochlear physiology has been to understand how sensory transduction operates at the basal end of the mammalian cochlea. The identification of candidate motor proteins in outer hair cells (OHCs) draws attention to the question of whether we understand cochlear tuning. Nevertheless, the association of the cloned motor protein 'prestin' with an anion transporter superfamily provides clues about the molecular nature of the OHC motor in the basolateral membrane, the utilisation of chloride in hair cells and the long-term stability of small basal turn cochlear hair cells. PMID- 11914518 TI - Prestin, the motor protein of outer hair cells. AB - Prestin is a gene recently cloned from mammalian cochlear outer hair cells (OHC) using a single cell type, outer minus inner hair cell, specific suppressive subtractive hybridization procedure. The localization and gene expression profile of the prestin protein fits the pattern of OHC's development of electromotility. When prestin is abundantly expressed in normally nonmotile kidney cells, nonlinear capacitance and motility that are normally only seen in OHCs can be recorded. Furthermore, both nonlinear capacitance and motility can be reduced by salicylate, a well-known inhibitor of electromotility. These data suggest that prestin is the motor protein of OHCs. Amino acid sequence and gene structure analysis indicate that prestin is the fifth member of a newly discovered anion transport family (SLC26) that includes PDS, DRA and DTDST, which are chloride iodide transporters, Cl-/HCO3- exchangers or sulfate transporters. Prestin shares overall structure similarity with this anion transporter family. Recently, intracellular anions (chloride or bicarbonate) were found to be essential for OHC electromotility and prestin's function. PMID- 11914519 TI - Mechanical transduction in outer hair cells. AB - The outer hair cells are responsible for the exquisite sensitivity, frequency selectivity and dynamic range of the cochlea. These cells are part of a mechanical feedback system involving the basilar membrane and tectorial membrane. Transverse displacement of the basilar membrane results in relative motion between the tectorial membrane and the reticular lamina, causing deflection of the stereocilia and modulation of the open probability of their transduction channels. The resulting current causes a change of membrane potential, which in turn produces mechanical force, that is fed back into the motion of the basilar membrane. Experiments were conducted to address mechanical transduction mechanisms in both the stereocilia and the basolateral cell membrane, as well as modes of coupling of the outer hair cell force to the organ of Corti. PMID- 11914520 TI - Non-linear aspects of outer hair cell transduction and the temporary threshold shifts after acoustic trauma. AB - The 200-Hz cochlear microphonic potential (CM) and the compound action potential (CAP) of the auditory nerve evoked by tone-bursts were recorded in the basal turn of the cochlea of anaesthetised guinea pigs, before and after exposure to traumatic high-frequency tones that produce a temporary threshold shift (TTS) in this cochlear region. The drop in CM and the TTS were highly correlated, suggesting that it is the disruption of the outer hair cells generating the CM that causes the TTS. The previously measured rise in endocochlear potential and drop in organ of Corti K+ levels suggest that the TTS is due to a temporary closure of outer hair cell mechanoelectrical transduction (MET) channels, which produces a drop in the mechanical sensitivity of the organ of Corti, due to disruption of the active process provided by outer hair cells. The time course of the onset and recovery of TTS is consistent with a kinetic folding and refolding of MET channels over a time course of hours and days. Mathematical modelling of this putative channel folding suggests that TTS recovery may be accelerated by the presentation of additional sounds during the recovery period. We present electrophysiological data (CM and CAP measurements) showing that this accelerated recovery occurs. Using two-tone complexes (phase-locked 5- and 10-kHz traumatic tones, and 10-kHz traumatic tones with 25-Hz bias tones), we also show that the mechanisms producing TTS are non-linear and asymmetric, and that the greatest 'trauma' occurs when the hair bundles of outer hair cells are deflected away from the basal body of these cells (i.e. in the direction normally causing hyperpolarisation of the cell membrane potential). PMID- 11914522 TI - Exploring the living cochlea using confocal microscopy. AB - To obtain a more integrated view of the cellular behaviour of the cochlea it is essential to observe not only wider regions of the exposed turns but also to visualize structures below the reticular lamina. Using confocal microscopy and in vitro preparations of guinea pig and mouse inner ears, cellular structures within the intact organ of Corti can be visualized at high resolution. The approach thus offers a means to investigate detailed cellular events, e.g. structural reorganization following acoustic overstimulation. Confocal microscope images, although sharper than images acquired using regular light microscopy, are still subject to problems related to light scattering within the optical system and low signal-to-noise ratio. Significant image restoration can, however, be obtained by applying a combination of wavelet denoising techniques and deconvolution algorithms. Future work will focus both on more dynamical cellular events and on new in vivo models where the inner ear is visualized at a better functional state. PMID- 11914521 TI - In vivo micromechanical measurements of the organ of Corti in the basal cochlear turn. AB - Cochlear mechanical measurements of organ of Corti motion are generally accomplished in the apical or basal turn as in vivo or in vitro studies. In the apex it is possible to observe and measure tectorial membrane vibration as well as vibrations of structures such as the reticular lamina or the basilar membrane (BM). However, compared to the basal turn, cochlear amplification and nonlinearity are not strong in the apex. Basal turn studies have typically been limited to point location measurements of the BM but improved technology for laser interferometry is now making possible the spatial mapping of BM motion. The 'complexity' of BM motion in the radial direction (particularly the phase variation) is important to new models of cochlear wave amplification. In future work it may be possible to learn about vibration of structures within the organ of Corti. PMID- 11914523 TI - Mechanoelectrical and voltage-gated ion channels in mammalian vestibular hair cells. AB - Mammalian vestibular afferents respond robustly to head movements at low frequencies and provide input to reflexes that control eye, head and body position. Vestibular organs have distinctive regions and hair cells: Type II cells receive bouton afferent endings and type I cells receive large calyx afferent endings. In the rodent utricle, type II cells are broadly tuned to frequencies between 10 and 30 Hz. Other recent data suggest that otolith organs function in this frequency range, which is higher than previously imagined. Some of the tuning derives from adaptation of the transducer current, which is best fitted with a double exponential decay with time constants of approximately 4 and 40 ms. Further tuning is provided by basolateral conductances, principally outwardly rectifying, voltage-gated K+ conductances. The kinetics of the K+ currents tend to vary with location in the sensory epithelium and therefore may contribute to regional variation in afferent physiology. Type I hair cells have a large, negatively activating K+ conductance, g(K,L), that confers a very low input resistance and therefore attenuates the receptor potential. This may reduce nonlinearity in the receptor potential, a possibly useful feature for the motor reflexes served by the vestibular system. On the other hand, the small receptor potentials together with unusually negative resting potentials are hard to reconcile with calcium-mediated quantal transmission. This problem may be overcome by factors that inhibit g(K,L)'s activation at resting potential. Also, the calyx may support nonquantal transmission. PMID- 11914524 TI - Roles of fibroblast growth factors in the inner ear. AB - The basic biology of the fibroblast growth factor (FGF) receptors and their splice variants is first reviewed, followed by a review of the known roles of FGFs in the inner ear. They include induction of the otocyst by FGF19, followed by FGF3 in further development of the otocyst. In later development, FGF3 or FGF10 acting on FGF receptor 2b is likely to be involved in development of the walls of the cochlear spaces, while FGF receptor 3 is involved in differentiation of the pillar cells of the organ of Corti. FGF1 and FGF2 act as trophic factors for the developing cochlear nerve fibres. PMID- 11914525 TI - The synaptic physiology of cochlear hair cells. AB - Mechanosensory hair cells of the vertebrate inner ear are so-called 'short' receptors that communicate to the central nervous system by way of chemical synapses with afferent neurons. In turn, hair cells are the targets of olivocochlear fibers that carry efferent inhibitory feedback from the brain. These synaptic activities contribute to, or modulate the hair cell's receptor potentials through the gating of associated ion channels. Thus for example, voltage-gated calcium channels open to trigger vesicle fusion and release of transmitter by entry of extracellular calcium. The inward calcium current also depolarizes the membrane and could lead to generation of 'all-or-none' action potentials. However, regenerative depolarization is prevented in most hair cells by prominent voltage-gated potassium conductances that rapidly repolarize the membrane. The magnitude and speed of these delayed potassium conductances determine the size and shape of the resulting receptor potential, and subsequent transmitter release, produced by sound. Efferent feedback is provided by the release of acetylcholine (ACh) from olivocochlear nerve fibers onto outer hair cells in the mammalian cochlea. The hair cell's ACh receptors are ligand-gated cation channels related to the nicotinic receptors of nerve and muscle. Calcium influx through the ACh receptors activates nearby calcium-gated potassium channels, resulting in hyperpolarization and inhibition of the hair cell. Calcium influx during efferent inhibition is regulated by a 'synaptic cistern' that also may act as a calcium store that is triggered by ACh under some conditions. PMID- 11914526 TI - Vesicle targeting in hair cells. AB - The mammalian hair cell has two distinct plasma membrane domains separated by tight junctions, the apical domain which contains the stereocilia and the basolateral domain which contains the presynaptic region. Little is known concerning the mechanisms that regulate vesicle trafficking to these two domains. Using SNAP 25 and syntaxin as baits, we carried out a yeast two-hybrid screen of the organ of Corti. We identified a novel syntaxin interacting protein, ocsyn, that is enriched in inner hair cells and concentrated at the apical pole. Our results are consistent with ocsyn playing a role in vesicle trafficking to the apical membrane of the hair cell. PMID- 11914528 TI - Purinergic regulation of sound transduction and auditory neurotransmission. AB - In the cochlea, extracellular ATP influences the endocochlear potential, micromechanics, and neurotransmission via P2 receptors. Evidence for this arises from studies demonstrating widespread expression of ATP-gated ion channels (assembled from P2X receptor subunits) and G protein-coupled receptors (P2Y receptors). P2X2 receptor subunits are localized to the luminal membranes of epithelial cells and hair cells lining scala media. These ion channels provide a shunt pathway for K+ ion egress. Thus, when noise exposure elevates ATP levels in this cochlear compartment, the K+ conductance through P2X receptors reduces the endocochlear potential. ATP-mediated K+ efflux from scala media is complemented by a P2Y receptor G protein-coupled pathway that provides coincident reduction of K+ transport into scala media from the stria vascularis when autocrine or paracrine ATP signalling is invoked. This purinergic signalling likely provides a basis for a reactive homoeostatic regulatory mechanism limiting cochlear sensitivity under stressor conditions. Elevation of ATP in the perilymphatic compartment under such conditions is also likely to invoke purinergic receptor mediated changes in supporting cell micromechanics, mediated by Ca2+ influx and gating of Ca2+ stores. Independent of these humoral actions, ATP can be classified as a putative auditory neurotransmitter based on the localization of P2X receptors at the spiral ganglion neuron-hair cell synapse, and functional verification of ATP-gated currents in spiral ganglion neurons in situ. Expression of P2X receptors by type II spiral ganglion neurons supports a role for ATP as a transmitter encoding the dynamic state of the cochlear amplifier. PMID- 11914527 TI - The inner hair cell synaptic complex: physiology, pharmacology and new therapeutic strategies. AB - Within the cochlea, the sensory inner hair cells (IHCs), which transduce mechanical displacement of the basilar membrane into neural activity, release glutamate to act on postsynaptic receptor channels located on dendrites of primary auditory neurons. In turn the activity of the postsynaptic auditory dendrites is modulated by a variety of lateral efferent neurotransmitters. This presentation reviews the most recent findings obtained at the IHC synaptic complex with an original technique, namely coupling auditory nerve single unit recordings with multibarrel intracochlear perfusions. Two types of results are emphasized: (1) in physiological conditions, the activity of auditory nerve fibers involves AMPA, but not kainate or NMDA receptors, and (2) this activity is tonically modulated by dopamine, one of the lateral efferent neurotransmitters. With the increasing knowledge of molecular mechanisms involved at the first synaptic complex in the cochlea, it is now possible to envisage local treatments for spiral ganglion neurons. These treatments, available experimentally, may be used in the near future: either to protect spiral ganglion neurons against excitotoxic injury (traumatic and/or ischemic sudden deafness), or to prevent excitotoxic-induced hyperexcitability (probably the starting point of most posttraumatic tinnitus), or to delay neuronal death (neural presbycusis). PMID- 11914529 TI - A role for purinergic receptors at the inner hair cell-afferent synapse? AB - Previously published evidence is reviewed for a functional role of ATP and its receptors at the chemical synapse between the mammalian inner hair cell and the primary afferent dendrites of the VIIIth nerve. New findings are also presented, both from gross cochlear potentials and single neurone recordings during intracochlear perfusion of ATP-receptor antagonists and agonists. Both the previous and present results are consistent with the notion that endogenous ATP may act to regulate the excitability of the primary afferent dendrite, possibly acting on P2X2 receptors. PMID- 11914530 TI - Cerebrovascular brainstem diseases with isolated cranial nerve palsies. AB - There is a significant number of individual patients with cranial nerve palsies as the sole manifestation of MRI- and, less frequently, CT-documented small brainstem infarctions or hemorrhages. The 3rd and 6th nerves are most commonly involved and, less frequently, the 4th, 5th, 7th, and 8th nerves. An intra-axial basis for such lesions may be underestimated if the diagnosis is based solely on MRI. The electrophysiologic abnormalities indicating brainstem lesions may be independent of MRI-documented morphological lesions. This paper reviews the literature on cerebrovascular brainstem diseases manifesting as isolated cranial nerve palsies. It supports the concept that small pontine and mesencephalic infarctions are the main cause of non-traumatic cranial nerve palsies in the middle-aged and elderly population. Microvascular infarction of the respective extra-axial cranial nerve segments seem to be less important. PMID- 11914531 TI - Antiphospholipid antibodies in the acute phase of cerebral ischaemia in young adults: a descriptive study of 139 patients. AB - A total of 139 young stroke patients were consecutively examined and tested for antiphospholipid antibodies (APLA) to evaluate the role of these antibodies in cerebral ischaemia before the age of 45. APLA were found in 28.8% of patients. Two factors, hypertriglyceridaemia and alcohol abuse, were significantly more frequent in patients with a positive APLA test. The demographic characteristics, other risk factors, history of prior thrombotic events and distribution of aetiopathogenic types of cerebral ischaemia were not different in patients with or without APLA. Laboratory assays for APLA were highly positive for only two patients, who both had autoimmune diseases. These results suggest that with the exception of a clinical context of antiphospholipid syndrome or other autoimmune diseases, the usefulness of this diagnostic tool in the management of cerebral ischaemia remains limited. PMID- 11914532 TI - Thrombolytic therapy in acute ischaemic stroke. AB - BACKGROUND: The aim of this study was to assess the feasibility, clinical effect and safety of intravenous thrombolysis with tissue plasminogen activator in patients with acute ischaemic stroke treated in an acute stroke unit. METHODS: All patients admitted within 3 h after an acute ischaemic stroke were considered for thrombolysis. Twenty-four patients were treated. RESULTS: Ten patients demonstrated early clinical improvement compatible with a positive effect of thrombolysis. Five patients demonstrated a substantial but slow clinical improvement with an uncertain relationship to thrombolysis. Nine patients did not improve. One patient developed an intracerebral haematoma and 2 developed a haemorrhagic infarction without clinical deterioration. Five patients (21%) died within the first 3 months. At follow-up after 6 months, 10 patients (42%) had achieved independence [modified Rankin Scale (mRS) 0-2], 9 (33%) had an unfavourable outcome (mRS 3-5) and 5 patients (21%) had died. None of these 5 patients died due to a treatment complication. CONCLUSIONS: This study in a small patient population suggests that thrombolysis may be administered relatively safely in an acute stroke unit without intensive care facilities. The clinical effect and safety were similar to those which have been found in large randomised studies and clinical series. PMID- 11914533 TI - Sonographic characterisation of radiation-induced carotid artery stenosis. AB - BACKGROUND AND PURPOSE: To study the distribution, extent and sonographic characterisation of radiation-induced carotid artery stenosis in nasopharyngeal carcinoma (NPC) patients. METHODS: The distribution of plaques, the extent of stenosis, and the sonographic characterisation of the plaque at maximum stenosis were recorded in 71 NPC patients. The results were compared with the ultrasound results of a control group of 142 patients presenting with symptoms of cerebrovascular disease or carotid bruit. RESULTS: NPC patients had a higher incidence of carotid stenosis (77 vs. 50.7%). The common carotid arteries were most commonly affected by radiation-induced stenosis (93/142 vs. 37/284 in the control group), whereas the carotid bulb was the most commonly affected (56/284) site in the control group. Significantly more NPC patients had moderate-to-severe stenosis (21/71 vs. 27/142). Analysis of the sonographic appearance of radiation induced and atherosclerotic plaques showed more diffuse involvement in the post radiation group. Non-calcified plaques and intraplaque hypoechoic foci were also more frequent in the post-radiation group. CONCLUSIONS: Radiation-induced carotid stenosis is more diffuse in distribution, is associated with more severe luminal stenosis and has different sonographic plaque characterisation compared with carotid stenosis without radiation exposure. PMID- 11914534 TI - Transoesophageal echocardiography in patients without arterial and major cardiac sources of embolism: difference between stroke subtypes. AB - We studied the records of 175 consecutive patients referred to our neurologic ward between January 1994 and February 2000 with a diagnosis of ischaemic cerebrovascular disease (ICVD) (stroke or transient ischaemic attack - TIA) who underwent transoesophageal echocardiography (TEE). We excluded patients with large vessel disease, high-risk embolic cardiopathies and other rare causes of stroke. According to clinical and neuroimaging findings, patients were divided into two groups. The lacunar (LAC) group (69/175 (39.4%)) and the nonlacunar (N LAC) one (106/175 (60.6%)). The control population consisted of 78 consecutive patients, referred to the echocardiography laboratory for TEE without history of ICVD and known heart disorders. Patent foramen ovale (PFO) frequency was significantly higher in case patients than in control subjects (55/175 (31.4%) vs. 13/78 (16.6%); p = 0.02). Among case patients, PFO was more prevalent in the N-LAC group than in the LAC one (43/106 (40.6%) vs. 12/69 (17.4%); p = 0.0005). A large degree of shunt occurred in 53.5% of N-LAC patients and in 16.7% of LAC ones (p = 0.04). Atrial septal aneurysm (ASA) was detected in 12% of case patients and 1.3% of control subjects (p = 0.003) and was more frequent in the N LAC group than in the LAC one (16 vs. 5.8%; p = 0.05). Mitral prolapse (MP) was present in 6/175 (3.4%) ICVD patients (vs. 1/78 among controls) in most cases associated with myxomatous valve redundancy. Aortic arch atheromas (AA) were detected in 12% of ICVD patients and in 10.2% of controls. The frequency was 9.4% in N-LAC and 15.9 in LAC. No complicated AA (plaque thickness >4 mm, ulcerated atheroma, superimposed thrombus) were detected. After multivariate analysis, PFO (OR = 3.8; 95% CI = 2.7-7.9) and ASA (OR = 8.01; 95% CI = 3.0-16.1) appeared to be independent predictors of ICVD. PFO (OR = 2.24; 95% CI = 1.24-4.92) was also independently associated with N-LAC stroke subtype and its importance was even higher in younger patients. Our study provides further evidence that TEE is a helpful diagnostic tool in stroke patients without arterial and major cardiac sources of embolism. However, its utility differs according to type and localization of the ischaemic lesion being more relevant in patient with N-LAC infarctions. PMID- 11914535 TI - Validity of the aphasia item from the Scandinavian Stroke Scale. AB - We studied the validity of the aphasia item of a widely used stroke scale - the Scandinavian Stroke Scale (SSS) - in discriminating between aphasia and normal language function in 33 stroke patients of an acute stroke unit. They were assessed by a nurse using the aphasia item from the SSS and by a speech and language therapist carrying out a full evaluation of the language function. The latter served as the 'gold standard'. The agreement between the nurses' and the speech and language therapist's scoring was good (weighted kappa = 0.74, 95% CI 0.51-0.97), and the sensitivity and specificity of the SSS aphasia item were also satisfactory. However, the predictive value of a positive test was as low as 0.55 (95% CI 0.23-0.83), indicating nearly every second of the positives being false positive. Using the aphasia score of the SSS as a diagnostic aid for aphasia after stroke results in a high rate of false positives and inflates the prevalence figures for aphasia in epidemiological studies of stroke. PMID- 11914536 TI - Altered food preference after cortical infarction: Korean style. AB - BACKGROUND: Altered food preference or preoccupation with certain food after stroke has been rarely described in the literature. METHODS: We report four Korean patients who developed altered food preference secondary to unilateral cortical infarction. RESULTS: Two patients showed preoccupation with meat such as Pulgogi or Kalbi (roast beef flavored with ingredients), which had not been their preference prior to the occurrence of stroke. All the patients became intolerant to the smell and taste of mackerel, and two disfavored hot Kimchi (cabbage salted with hot pepper). Quantitative taste assessment done in the patients revealed decreased taste perception in a variable pattern along with dysgeusia. Imaging studies showed that the region commonly involved was the frontal opercular area corresponding to the taste cortex. CONCLUSION: It is suggested that strokes involving the taste cortex may produce altered food preference associated with decreased taste sensation. PMID- 11914537 TI - Neurological outcome and quality of life after stroke due to vertebral artery dissection. AB - BACKGROUND: Vertebral artery dissection is a well-recognized cause of posterior circulation stroke for which there is relatively little information on long-term outcomes. Quality of life (QOL) is an important patient-centred outcome measure. METHODS: Stroke due to vertebral artery dissection was conservatively defined by neuroimaging documentation. Thirty sequential cases were identified based on a retrospective database and chart review with prospective follow-up. Surviving patients completed the Short Form-36 (SF-36) and the Stroke-Specific Quality of Life (SSQOL) scales and were subsequently examined neurologically and scored on the National Institutes of Health Stroke Scale (NIHSS). Comparisons were made between outcome on the stroke scale and QOL scales and between outcome on the SF 36 and the Canadian population. RESULTS: There was discordance between outcomes recorded on a standard stroke scale and QOL measures with more patients scoring poorly on QOL measures. QOL was low in one third of the survivors. Overall QOL was significantly lower than the general population. CONCLUSIONS: Stroke due to vertebral artery dissection results in poorer outcomes on patient-centred QOL measures than on a standard stroke scale. PMID- 11914538 TI - Inflammatory leukocyte infiltration in focal cerebral ischemia: unrelated to infarct size. AB - OBJECTIVE: An inflammatory host response in the ischemically injured brain is well documented. However, its pathophysiological relevance is uncertain. We investigated whether inflammatory leukocyte response in the ischemic brain alters infarct size. METHODS: The cellular inflammatory response to cerebral ischemia in Wistar-derived rats induced by the transient occlusion of the middle cerebral artery with a thread was pharmacologically upmodulated by lipopolysaccharide (LPS) or downmodulated by continuous infusion of carboxylated sialyl Lewis(x) (sLex). The effects of such experimental modulation of focal cerebral leukocyte recruitment on the extent of the resulting infarction were assessed. RESULTS: Compared to control treatments, LPS strongly enhanced (540.5 +/- 504.8 vs. 94.6 +/- 60.6, p < 0.01) and sLex decreased (32.8 +/- 29.1 vs. 97.0 +/- 49.7, p < 0.05) the numbers of neutrophils at the investigated sites in cerebral ischemia. Unexpectedly, despite such marked experimental modulation of leukocyte infiltration in the ischemic brain, the extent of the resulting cerebral infarction (percent of total hemisphere) remained unchanged under these different conditions (54.5 +/- 10.8 vs. 53.0 +/- 19.1, n.s. and 50.3 +/- 18.0 vs. 57.2 +/- 10.0, n.s., respectively). CONCLUSIONS: The striking dissociation between the massively altered inflammatory leukocyte infiltration in the ischemic brain and the unchanged infarct outcome indicates that intracerebral inflammatory leukocyte recruitment is not a major pathogenic factor in the development of ischemic tissue damage. PMID- 11914539 TI - Blood pressure in acute stroke. The Copenhagen Stroke Study. AB - This study examines blood pressure (BP) and independent factors related to BP in the acute phase of stroke. The study is part of the community-based Copenhagen Stroke Study. In a multivariate regression model we analyzed the impact of clinical and medical factors on admission BP. BP declined with increasing time from stroke onset with a total of 8/4 mm Hg. Independent factors related to diastolic BP were ischemic heart disease (-3.9 mm Hg), male gender (2.2 mm Hg), known hypertension prior to stroke (8.6 mm Hg), and primary hemorrhage (9.7 mm Hg). Independent factors related to systolic BP were age (3.6 mm Hg/10-year increase), atrial fibrillation (-7.2 mm Hg), ischemic heart disease (-6.0 mm Hg), intracerebral hemorrhage (13.3 mm Hg), and known hypertension prior to stroke (16.3 mm Hg). No independent relations were seen between BP and diabetes, claudication, previous stroke, smoking, daily alcohol consumption, initial stroke severity and lesion size. The increase in BP in the acute phase of stroke is a uniform response to the ischemic event per se. BP is not related to stroke severity. Several factors are independently related to the BP level in acute stroke. The clinical significance of this is yet to be tested, but these factors may contribute to the seemingly complex relation between BP and outcome. PMID- 11914540 TI - Stroke following chiropractic manipulation. Report of 3 cases and review of the literature. AB - We present 3 cases of stroke due to arterial dissection following chiropractic manipulation: (1) a 31-year-old woman with left vertebral dissection developed a large cerebellar infarct, (2) a 64-year-old man developed a left parietal infarct due to left carotid dissection and (3) a 51-year-old man developed right Horner's syndrome, fluctuating dysarthria, left facial droop, and left arm weakness due to right carotid dissection. Imaging studies and the literature are reviewed. PMID- 11914541 TI - Extensive cerebral venous thrombosis associated with moderate hyperhomocyst(e)inemia and successfully treated with thrombolysis. AB - Cerebral venous thrombosis (CVT) is a relatively rare condition caused by multiple etiologies, usually secondary to hypercoagulable states. The optimal treatment is controversial, especially in the presence of hemorrhagic infarcts. We describe an unusual patient with extensive CVT associated with moderate hyperhomocyst(e)inemia who was aggressively treated with chemical and mechanical thrombolysis with an excellent outcome. PMID- 11914542 TI - Facio-oculo-palatal myoclonus due to pontine cavernous angioma. PMID- 11914543 TI - Intracellular cysteines of the cystic fibrosis transmembrane conductance regulator (CFTR) modulate channel gating. AB - The cystic fibrosis transmembrane conductance regulator (CFTR), a member of the ATP-binding cassette superfamily, is a cAMP-activated chloride channel. CFTR contains two transmembrane domains (TMDs), two nucleotide-binding domains (NBDs), and a regulatory (R) domain. We found that whole-cell CFTR-dependent Cl- currents in Xenopus laevis oocytes were sensitive to HgCl(2), suggesting that modification of endogenous cysteines alters channel activity. To understand better this phenomenon, site-directed mutagenesis was employed to generate both individual cysteine replacements and a version of the molecule with no cysteines in the hydrophobic sector. Each mutant displayed a forskolin/IBMX-activated Cl(-) conductance similar to wild type, indicating that none of the cysteines located within the TMDs is essential. Subsequent single-channel analysis of inside-out patches excised from HEK293 cells expressing either cysteine-less or wild-type CFTR showed that intracellular application of a membrane impermeant sulphydryl reagent, p-chloromercuribenzosulfonate (PCMBS), significantly reduced open probability without affecting ion selectivity or conductance. The cysteine-less molecule also acquired a voltage-dependent sensitivity to extracellular PCMBS not observed in the wild type, perhaps due to a more flexible conformation that allowed PCMBS access to the intracellular surface. Together, these experiments suggest that endogenous intracellular cysteines, located primarily within the NBDs and/or R domain, influence channel gating. PMID- 11914544 TI - Calmodulin-mediated regulation of bicarbonate and lactate transports in rat jejunum. AB - BACKGROUND/AIMS: Ca(2+)/CaM is known to modulate the activity of several transport systems and its regulation can be accomplished either directly or via the involvement of specific protein kinases. Aim of this study was to investigate the possible role of Ca(2+)/CaM on bicarbonate and lactate transports in rat jejunal enterocyte. METHODS: Enzymatic assays in isolated plasma membranes were performed. Moreover membrane vesicles, transiently opened and resealed, were loaded with Ca(2+) and calmodulin, both in the absence and in the presence of ATP, and were used after that to perform uptake studies. RESULTS: Enzymatic assays gave evidence for the presence of Ca(2+)/CaM-dependent protein kinase II (CaMKII) in plasma membranes from rat jejunum. However, uptake experiments suggest that Ca(2+)/CaM, and not CaMKII, inhibits both basolateral Cl(-)/HCO(3)( ) exchange and H(+)-lactate symport, whilst HCO(3)(-) and Cl(-) conductances are unaffected. Neither Ca(2+)/CaM nor CaMKII seem to regulate brush border Na(+)/H(+) exchanger activity. CONCLUSION: These data are consistent with a Ca(2+)/CaM-mediated reduction of bicarbonate and lactate exit from jejunal enterocyte. PMID- 11914545 TI - Simvastatin inhibits malignant transformation following expression of the Ha-ras oncogene in NIH 3T3 fibroblasts. AB - In previous studies we have shown that the expression of the transforming Ha-ras oncogene in NIH 3T3 fibroblasts stimulates cellular calcium entry, which triggers oscillatory calcium induced calcium release from internal stores. The intracellular calcium oscillations lead to cytoskeletal remodeling by actin stress fiber depolymerization and activation of the Na(+)/H(+) exchanger thus mediating cell swelling and intracellular alkalosis, both important mitogenic signals. This is evidenced by abrogation of Ha-ras induced growth factor independent cell proliferation by interference with any of these events, i.e. by inhibition of cellular calcium entry or inhibition of the Na(+)/H(+) exchanger. As shown in this study, simvastatin, an inhibitor of 3-hydroxy-3-methylglutaryl coenzyme A (HMG-CoA) reductase, the key enzyme for cholesterol biosynthesis, is able to prevent these events following the expression of the transforming Ha-ras oncogene. We show, that simvastatin inhibits farnesylation dependent membrane translocation of a CAAX motive bearing yellow fluorescent protein and suppresses Ha-ras stimulated cellular calcium influx, which can be identified as capacitative calcium entry. In addition simvastatin is able to block regulatory volume decrease channels and to suppress the cytoskeletal remodeling, intracellular alkalinization, increase in cell volume and growth factor independent cell proliferation induced by the oncogene. Thus simvastatin is able to prevent crucial cellular events following expression of the transforming Ha ras oncogene. PMID- 11914546 TI - Activation of human somatostatin receptor 2 promotes apoptosis through a mechanism that is independent from induction of p53. AB - The ability of both somatostatin (SS) and its stable analogues to inhibit cell growth depends on the stimulation of specific membrane receptors (SSTR1-5), which belong to the G protein-coupled receptor family. Accumulating evidence suggests that the SSTR2 plays a major role in mediating cell cycle arrest, and it is also clear that SHP-1, a cytoplasmic phosphotyrosine phosphatase (PTP), is an essential component of the SSTR2-mediated cytostatic effect. In contrast, the possibility that SSTR2 activation may also lead to increased apoptosis is still beyond debate, despite SHP-1 activation is also able to promote cell death in several cell types. In the present work we have investigated the ability of SSTR2 to induce apoptosis in HL-60 cells. We have found that HL-60 cells uniquely express the SSTR2 subtype, and that stimulation of SSTR2 with the SS analogue SMS 201-995 results in an increased cell death. In all, these findings demonstrate that activation of SSTR2 promotes apoptosis in HL-60 cells. Moreover, in contrast with the proapoptotic mechanism previously reported for SSTR3, cell death induced by activation of SSTR2 is independent from accumulation of p53. PMID- 11914547 TI - Low-flow ischemia and hypoxia stimulate apoptosis in perfused hearts independently of reperfusion. AB - Post-ischemic reperfusion leads to apoptosis-linked loss of myocytes in cultured cells and in vivo. We tested the hypothesis that apoptosis develops without reperfusion in Langendorff-perfused hearts exposed to either low-flow ischemia (LFI) or hypoxia (H). Rat hearts were perfused with amino-acid-enriched Krebs Henseleit buffer and exposed for 6 h to LFI (flow=2 ml/min, PO(2)=500+/-50mmHg, mean+/-SD), H (10ml/min, 120+/-15mmHg), or control conditions (C, 10ml/min, 500+/ 50mmHg). At selected times, DNA-fragmentation was measured by agarose-gel electrophoresis and in situ TUNEL assay. After 6 h, the ratio (TUNEL positive)/(total nuclei) was 0.620+/-0.027, 0.615+/-0.005, 0.404+/-0.021 in LFI, H and C, respectively. The ratio was 0.813+/-0.021 in hearts exposed to 90 min global no-flow ischemia and reperfused (5 h). To assess the role of membrane diffusible factors, separate experiments were performed recirculating the medium and exposing hearts to LFI or H as above. The degree of apoptosis was the same in both the recirculating and non-recirculating modes. Thus, apoptosis develops by similar extents and in a time-dependent fashion in crystalloid-perfused rat hearts during LFI or H at the same oxygen shortage (flow.PO(2)), even without the reperfusion. PMID- 11914548 TI - Expression of the serine/threonine kinase hSGK1 in chronic viral hepatitis. AB - The human serine/threonine kinase hSGK1 is expressed ubiquitously with highest transcript levels in pancreas and liver. This study has been performed to determine the hSGK1 distribution in normal liver and its putative role in fibrosing liver disease. HSGK1-localization was determined by in situ hybridization, regulation of hSGK1-transcription by Northern blotting, fibronectin synthesis and hSGK1 phosphorylation by Western blotting. In normal liver hSGK1 was mainly transcribed by Kupffer cells. In liver tissue from patients with chronic viral hepatitis, hSGK1 transcript levels were excessively high in numerous activated Kupffer cells and inflammatory cells localized within fibrous septum formations. HSGK1 transcripts were also detected in activated hepatic stellate cells. Accordingly, Western blotting revealed that tissue from fibrotic liver expresses excessive hSGK1 protein as compared to normal liver. TGF beta1 (2 ng/ml) increases hSGK1 transcription in both human U937 macro-phages and HepG2 hepatoma cells. H(2)O(2) (0.3 mM) activated hSGK1 and increased fibronectin formation in HepG2 cells overexpressing hSGK1 but not in HepG2 cells expressing the inactive mutant hSGK1(K127R). In conclusion hSGK1 is upregulated by TGF-beta1 during hepatitis and may contribute to enhanced matrix formation during fibrosing liver disease. PMID- 11914549 TI - Detection of microembolic signals in patients with neuropsychiatric lupus erythematosus. AB - The pathogenesis of central nervous system involvement in systemic lupus erythematosus (SLE) is not completely understood. In this study, we investigated the association of microembolic signals (MES) with a variety of neuropsychiatric SLE manifestations and compared our results with those from SLE patients without neuropsychiatric lupus and normal controls. Fifty-three patients with SLE (45 females and 8 males), all fulfilling the revised classification criteria for SLE, and 50 control subjects (44 females and 6 males) were enrolled in this study. All SLE patients were assessed by neuropsychological examination, including various neuropsychiatric tests. Twenty-five patients with SLE were found to have at least one of the neuropsychiatric syndromes defined by The American College of Rheumatology. The mean MES count in patients with neuropsychiatric lupus was significantly higher than those without (5.4 +/- 1.1 vs. 0.3 +/- 0.8/h; p < 0.005). We found a positive correlation between higher mean MES counts and the presence of neuropsychiatric syndromes in SLE. The mean MES count in the whole group of SLE patients was also significantly higher than that in healthy controls. The mean MES count of SLE patients with antiphospholipid (aPL) antibody positivity was significantly higher than those without aPL antibodies (3.6 +/- 1.6 vs. 0.8 +/- 0.1/h; p < 0.005). In conclusion, the association of MES with neuropsychiatric lupus may support the possible contribution of MES to the complex pathophysiology of this syndrome. More importantly, detection of MES on transcranial Doppler monitoring might suggest a high risk of involvement of the central nervous system in SLE, and could be used as a diagnostic tool. PMID- 11914550 TI - Dystypia: isolated typing impairment without aphasia, apraxia or visuospatial impairment. AB - We report a 60-year-old right-handed Japanese man who showed an isolated persistent typing impairment without aphasia, agraphia, apraxia or any other neuropsychological deficit. We coined the term 'dystypia' for this peculiar neuropsychological manifestation. The symptom was caused by an infarction in the left frontal lobe involving the foot of the second frontal convolution and the frontal operculum. The patient's typing impairment was not attributable to a disturbance of the linguistic process, since he had no aphasia or agraphia. The impairment was not attributable to the impairment of the motor execution process either, since he had no apraxia. Thus, his typing impairment was deduced to be based on a disturbance of the intermediate process where the linguistic phonological information is converted into the corresponding performance. We hypothesized that there is a specific process for typing which branches from the motor programming process presented in neurolinguistic models. The foot of the left second frontal convolution and the operculum may play an important role in the manifestation of 'dystypia'. PMID- 11914551 TI - Clinical aspects of the AIDS dementia complex in relation to histopathological and immunohistochemical variables. AB - To correlate cerebral histopathological and immunohistochemical changes in the neuroclinical features of the AIDS dementia complex (ADC), autopsy results of 28 ADC patients were related, in a retrospective analysis, to scores on a standardised neurological examination performed at neurologic onset. From a histopathological point of view, the cases were classified as follows: 9 cases of HIV leucoencephalopathy (HIVL; diffuse myelin damage and rare microglial nodules), 7 cases of HIV encephalitis (HIVE; several microglial nodules and no myelin damage) and 12 cases of mixed HIVL and HIVE (HIVL-E). The groups differed significantly with respect to symptoms and CD4 count at neurologic onset, survival and neurological impairment. Immunohistochemically, the interstitial component (p24-positive cells scattered singly within the white matter) was significantly more prevalent in HIVL, and the micronodular component (p24 positive cells confined within microglial nodules) in HIVE. Neurological damage was worse in cases with a high prevalence of interstitial component or a low prevalence of micronodular component. HIVE, HIVL and HIVL-E are distinct clinical forms of ADC. Neurological impairment is related to white matter damage. PMID- 11914552 TI - Is the right amygdala involved in visuospatial memory? Evidence from MRI volumetric measures. AB - Quantitative MRI measurement of hippocampal sclerosis in patients suffering from temporal lobe epilepsy (TLE) have, as yet, failed to evidence any correlation between the right hippocampus and visuospatial memory. In this report, word learning and design learning tasks were carried out as well as MRI volumetric measurements of the hippocampus and amygdala in order to verify possible modality specific correlations between function and structure. Delayed recall indices in our memory tasks provided significant results. Visuoverbal ratios differed between right and left TLE groups, as did laterality indices of hippocampal and amygdalar volumes. Furthermore, correlations were found between the left hippocampal volume and verbal memory, and between the right amygdala and visuospatial memory. We suggest that the difficulty encountered in establishing a correlation between right temporal structures and visuospatial memory could come both from the type of test employed and the structures considered. PMID- 11914553 TI - Severe personality changes after unilateral left paramedian thalamic infarct. AB - Personality changes are not uncommon after paramedian thalamic infarction, but usually bilateral or relatively large lesions, often complicated by other neurological or neuropsychological deficits, are present. 'Pure' cases of unilateral lesions are extremely rare. We report that a right-handed, 48-year-old man, who was hypertensive and diabetic but had no prior psychiatric history, developed severe personality changes and a frontal-like syndrome after recovery from acute-onset impairment of consciousness at the age of 43. Other neurological and neuropsychological disturbances, especially verbal and visual amnesia, were unremarkable. MRI showed a very small infarct in the left paramedian area of the thalamus, mainly involving the dorsomedial nucleus. PMID- 11914554 TI - Menopause and menarche in patients with primary blepharospasm: an exploratory case-control study. AB - We studied the relationships between blepharospasm (BSP) and menopause/menarche in female patients with primary BSP (n = 83) and age-matched healthy (n = 83) and disease controls (n = 83). BSP patients and matched controls had comparable age at menopause, and there was no correlation between age at menopause and age at BSP onset. Thus, menopause probably exerts no significant influence on the age dependent development of BSP. BSP cases tended to have a later menarche than either group of controls. The association was independent of age, disease duration and education level. Because the higher the age at menarche, the higher the age at BSP onset, later menarche was unlikely to be a risk factor for BSP. Rather, the two conditions may share pathophysiologic mechanisms, for example minor abnormality of neurotransmitter systems controlling both the motor system and the maturation of the hypothalamic-pituitary-gonadal axis responsible for the onset of puberty. PMID- 11914555 TI - High occurrence rate of glaucoma among patients with Alzheimer's disease. AB - The purpose of the current study was to reveal the occurrence rate of glaucoma among patients with Alzheimer's disease (AD). All 112 patients of four nursing homes in Upper Bavaria, Germany, who met the diagnostic criteria of probable AD, were incorporated into the study. Visual field defects and/or optic disc cupping compatible with the diagnosis of glaucoma were found in 29 out of 112 patients with AD (25.9%). When compared to a control group (5.2%) and to the prevalence of glaucoma in western countries revealed in a number of glaucoma surveys (2.6 4.7%), patients with AD may have a significantly increased occurrence rate of glaucoma. In addition, ocular hypertension with normal visual fields and normal optic nerve heads was not found in patients with AD. The prevalence of ocular hypertension in the control group was 7.8% and parallels previous surveys. Therefore, we assume that the optic nerve seems to be less resistant to elevated intraocular pressure levels in AD patients. PMID- 11914556 TI - MRI morphometric study and correlation with cognitive functions in young adults shunted for congenital hydrocephalus related to spina bifida. AB - We studied the morphometric MRI findings and their correlation with cognitive functions in a population of 10 young adults shunted for congenital hydrocephalus related to spina bifida. Morphometric MRI analysis included measurement of the ventricular dilatation index, frontal and occipital parenchymal thickness and the size of the corpus callosum. The neuropsychological status was evaluated, notably to look for a discrepancy between verbal and performance skills, a finding which has previously been described in hydrocephalic children. We also investigated whether there was a correlation between cognitive function and cerebral morphometric indexes. In each case, MRI demonstrated the structural changes associated with the Chiari II malformation. The size of the lateral ventricles varied, ranging from important dilatation to small ventricles. Six patients had only partial development of the corpus callosum. All patients had a normal global IQ. In our population of young adults, we did not observe any discrepancy between verbal and visuospatial performances as has been described in children with hydrocephalus. We found no relationship between cognitive function and ventricle dilatation or parenchymal thickness or between the size of the corpus callosum and callosal transfer. PMID- 11914557 TI - Focal neurological deficits following spontaneous thrombosis of unruptured giant aneurysms. AB - BACKGROUND: Giant aneurysms (GAs) must be regarded as dynamic lesions with respect to their growth and intraluminal thrombus formation, occasionally giving rise to thromboembolic events. Little is known about spontaneous thrombosis of unruptured GAs leading to focal neurological deficits which can mimic arteriosclerotic, epileptic or neoplastic disease. METHODS: Three patients (2 men, 27 and 68 years old, and 1 woman, 32 years old) presented with progressive neurological deficits. Neuroradiological studies showed unruptured GAs [two of the middle cerebral artery (MCA), one of the anterior communicating artery] displaying spontaneous near complete intraluminal thrombosis. In one of the cases with a giant MCA aneurysm, acute thrombosis was followed by infarction of the corresponding territory. RESULTS: Partial excision of the intraluminal thrombus was performed in the anterior communicating artery lesion. Wrapping followed by an extracranial-intracranial bypass and stepwise trapping successfully excluded one MCA lesion from circulation. The other MCA aneurysm was treated conservatively. All three patients showed full recovery neurologically after treatment. CONCLUSION: The reported clinical history of three patients who developed rapid focal progressive neurological deterioration following acute spontaneous thrombosis can be attributed to the acute swelling of the aneurysmal mass. Their pattern of consecutive rapid clinical improvement represents a previously undescribed clinical course of unruptured GAs. PMID- 11914558 TI - Paroxysmal focal dystonia in neuro-Behcet by a small ipsilateral thalamic lesion. PMID- 11914559 TI - Primary central nervous system lymphoma with Waldenstrom's macroglobulinaemia. PMID- 11914560 TI - Cervical spinal cord infarction simulating myocardial infarction. PMID- 11914561 TI - Cystic dilation of Virchow-Robin spaces in the midbrain. PMID- 11914562 TI - Early amniocentesis and congenital foot deformities. AB - OBJECTIVE: Several studies have shown an increased risk of congenital foot derformities after early amniocentesis. These studies have comprised amniocenteses performed before 13 completed gestational weeks. In this study, the risk of foot deformities after amniocentesis performed at 12-14 completed gestational weeks was determined. METHODS: 3,469 genetic amniocenteses in singleton pregnancies performed before 15 completed gestational weeks were studied. The intention was to perform the amniocenteses at 12-14 weeks of gestation, but 32 amniocenteses were performed at the gestational age 11 weeks + 5 days or 11 weeks + 6 days. The pregnancies were followed up with regard to fetal loss and leakage of amniotic fluid. After birth, newborns with a diagnosis of foot deformity were identified from the Swedish Medical Birth Registry. The observed number of foot deformities was then compared with the expected number which was calculated stratified for delivery hospital, year of birth, and maternal age. RESULTS: The observed number of foot deformities was significantly higher than the expected: exact odds ratio 1.74 (exact 95% confidence interval 1.06-2.69). The rate of spontaneous abortions after the procedure was 1.8%, and the rate of leakage of amniotic fluid was 1.9%. There was a significant trend for all complications to decrease with increasing gestational age at amniocentesis. CONCLUSION: Women undergoing amniocentesis at 11+5 to 14+6 gestational weeks have an increased risk of giving birth to a child with a congenital foot deformity. PMID- 11914563 TI - First case of trisomy 13 plus mosaic trisomy 1q. AB - OBJECTIVES: In a case with severe sonographic abnormalities followed by missed abortion in week 14 + 5 days, cytogenetic analysis was performed on placental tissue. A mosaic karyotype 47,XY,+13,add(1)(q44)[3]/47,XY, +13[9]/46,XY[36] was detected. The purpose of the present study was to characterize the additional material on chromosome 1q. METHODS: According to GTG banding, the additional material on chromosome 1 was most probably chromosome 1 material. Thus, multicolor banding analysis using a chromosome 1 specific probe set was done to precisely describe the rearranged chromosome 1. RESULTS: Molecular cytogenetic approaches revealed that the derivative chromosome 1 was der(1)(1p36.3- >1q44::1q12-->1q44). CONCLUSIONS: This is the first description of a case with a trisomy 13 plus a partial trisomy 1q presenting with clinical signs of both aberrations. Moreover, the multicolor banding technique is suited to resolve complex karyotypes in the prenatal diagnosis, i.e., chromosome preparations of fibroblasts. PMID- 11914564 TI - Second-trimester maternal pregnancy-associated plasma protein a and inhibin a levels in fetal trisomies. AB - OBJECTIVE: The aim of this study was to determine whether fetal trisomy is associated with altered levels of second-trimester maternal pregnancy-associated plasma protein A (PAPP-A) and inhibin A. METHODS: Maternal serum PAPP-A and inhibin A concentrations were measured at 15-17 weeks of gestation in 14 singleton pregnancies with fetal trisomy and in 56 matched pregnant controls. RESULTS: PAPP-A levels in the trisomy group were significantly lower than in controls. The inhibin A level with fetal trisomy 21 was slightly higher than the control group, but levels were not different between trisomies 18 and 13 and controls. CONCLUSION: Fetal trisomies 21, 18, and 13 are associated with a reduction in second-trimester maternal PAPP-A levels; trisomies 18 and 13 are not associated with increased inhibin A levels, unlike trisomy 21. PMID- 11914565 TI - Influence of volume preloading on uteroplacental and fetal circulation during spinal anaesthesia for caesarean section in uncomplicated singleton pregnancies. AB - OBJECTIVE: Effects of volume preloading during spinal anaesthesia for elective caesarean section on maternal blood pressure, feto-maternal circulation and fetal outcome. PATIENTS AND METHODS: In a pilot study a randomised trial was performed in 22 healthy women with uncomplicated, singleton pregnancies at 36-40 weeks of gestation undergoing elective caesarean section under spinal anaesthesia. In the low volume group (group A) patients received 150 ml of crystalloid solution for preloading, in the high volume group (group B) they were given 15 ml/kg of crystalloid solution for preloading before the initiation of spinal anaesthesia. Maternal blood pressure was monitored intermittently. Hypotension was defined as a decrease in systolic pressure to less than 80% of the baseline value. The Doppler flow evaluation consisted of measurements from the uterine artery at the placental site, fetal umbilical artery and fetal middle cerebral artery. Pulsatility indices were derived before and after fluid preloading, and when spinal anaesthesia was established. The neonatal outcome was assessed by Apgar scores, arterial acid base status and neurologic and adaptive capacity scores (NACS). RESULTS: The incidence of maternal hypotension in both groups was 45.5% (n = 10); 3 cases occurred in group A compared to 7 cases in group B (n.s.). There was no evidence that the high dose volume is useful in preventing maternal hypotension. The pulsatility indices of uterine arteries, umbilical arteries and middle cerebral arteries were not altered. Statistical analysis showed no changes in neonatal outcome concerning umbilical arterial pH, Apgar score and NACS (n.s.) between groups A and B. CONCLUSIONS: Our preliminary results suggest that high dose crystalloid volume preloading has no preventive function in the avoidance of maternal hypotension in healthy parturients undergoing elective caesarean section under spinal anaesthesia, and shows no harmful effects on neonatal outcome as long as maternal hypotension is corrected immediately. However, the statistical significance may reflect the small sample size, and larger series are needed before changing the current management. PMID- 11914566 TI - Does amniotic fluid alpha-fetoprotein have diagnostic or prognostic value at the time of second midtrimester genetic amniocentesis? AB - In order to assess the usefulness of amniotic fluid alpha-fetoprotein (AFP) levels at the time of midtrimester genetic amniocentesis, 4,430 cases were retrospectively studied to compare the high, normal or low AFP values with the karyotype characteristics and fetal anatomy using ultrasound (US) scanning and confirmed by postnatal evaluation or necroscopy in the case of termination of pregnancy. All the cases presenting malformations were correctly diagnosed by US examinations. AFP levels over the 2nd standard deviation (SD) were found in 112 cases (2.52%) and below the 2nd SD in 11 cases (0.24%). The characteristics of these cases are presented and discussed. According to our results, it is concluded that routine assessment of AFP at the time of midtrimester genetic amniocentesis, if coupled with optimal US scanning, is no longer justified. PMID- 11914567 TI - Arthrogryposis multiplex congenita and cerebellopontine ischemic lesions in sibs: recurrence of prenatal disruptive brain lesions with different patterns of expression? AB - Arthrogryposis multiplex congenita (AMC) is a heterogeneous group of disorders in which prolonged decrease or absence of fetal movements results in a series of deformational anomalies. The rate of recurrence ranges from 25% in some recessive forms of myogenic arthrogryposis or of primary anterior horn cell loss, to less than 1% in anoxic-ischaemic damage. Cerebral clastic processes are considered as sporadic. We report on a non-consanguineous family in which the first child was affected by AMC and the following pregnancy was terminated because cerebellum hypoplasia was suspected at ultrasound and confirmed by fetal magnetic resonance imaging. Post-mortem findings demonstrated pontocerebellar ischaemic-haemorrhagic injuries. The occurrence of these neurologic abnormalities in the same family suggests a common mechanism, which might correspond to a same genetic defect with different patterns of expression. This is the first prenatal report suggesting that an 'ischaemic' process, usually recognised as sporadic could in fact be due to an inherited abnormality. Careful prenatal follow-up of third-trimester fetal brain development may be required in pregnant women with a family history of AMC. PMID- 11914568 TI - Systemic diffusion including germ cells after plasmidic in utero gene transfer in the rat. AB - OBJECTIVES: The integration of a proviral plasmid into the host genome could be a good approach for fetal somatic gene transfer. The goal of this study was to assess the integration and transcription of a proviral marker gene injected intraperitoneally into rat fetuses as well as the risks of maternal contamination and germ-line transmission. METHODS: On day 17 post-coitus, each fetus was injected intraperitoneally with 10 microg plasmid DNA through the uterine wall. Twenty-one days after spontaneous delivery, integration and transcription of the plasmid in gonad, gut, liver, spleen, lung and brain tissue from 10 pups were determined by PCR and RT-PCR. RESULTS: 14 of 60 organs exhibited integration of the plasmid. Four samples of gut (40%), 3 samples of liver and spleen (30%), 2 samples of brain (20%) and no sample of lung were transfected. Two testicular samples were transfected and study of F1 rats from 2 brothers of one of the positive rats revealed transgenic pups from 1 of these 2 animals. No transfection of maternal tissues was detected. CONCLUSION: Integration and transcription of a marker gene injected intraperitoneally into rat fetuses appear efficient, especially in intraperitoneal organs. The risk of maternal contamination appears very low when using a naked DNA plasmid injected directly into fetuses. However, germ-line contamination can occasionally occur even with injection late during pregnancy, suggesting further studies are necessary to assess this risk in direct gene transfer experiments. PMID- 11914569 TI - Intervillous and spiral artery flows in normal pregnancies between 5 and 10 weeks of amenorrhea using color Doppler ultrasonography. AB - OBJECTIVE: The aim of our study was to investigate early placental circulation development and spiral and uterine artery flows in normal 1st-trimester pregnancies. METHODS: A prospective study of intervillous and spiral artery flows in 49 normal pregnancies (5-10 weeks of amenorrhea) was performed. Transvaginal color and pulsed-wave Doppler techniques (6-MHz probe) were used as routine ultrasound scanning modalities before pregnancy termination for psychosocial reasons. RESULTS: In all pregnancies, between 5 and 10 weeks, continuous nonpulsatile intervillous flow (mean V(max) 3.55 cm/s) and spiral artery flow (mean peak systolic velocity 16.2 cm/s, mean diastolic index D/S 0.49 +/- 0.089) were detected. In the 47 women in whom the uterine artery flow was measured, the mean peak systolic velocity was 67.5 cm/s, the mean diastolic index was 0.12, and bilateral notching was observed. CONCLUSIONS: In all cases, between 5 and 10 weeks, slow and continuous nonpulsatile intrachorionic flow could be detected, whereas pulsatile flow was detected in spiral arteries. The true nature of this early intervillous circulation remains to be determined. PMID- 11914570 TI - Soluble intercellular adhesion molecule 1 in umbilical cord serum: potential for the diagnosis of neonatal infections. AB - OBJECTIVE: To evaluate the diagnostic relevance to neonatal infections of the soluble intercellular adhesion molecule 1 (sICAM-1) cord serum level. METHODS: The case-control study included 66 term newborn infants with and without risk factors for neonatal infections. Cord blood serum determinations of white blood cell count, C-reactive protein, fibrinogen, and sICAM-1 were systematically performed associated with bacterial cultures from placenta, ears, and gastric fluids. RESULTS: 6 of 33 infants (18.2%) with risk factors were infected, and 13 (39.4%) were colonized. Two infants included in the group without infection risk factors (n = 33) were colonized. No difference in sICAM-1 cord serum levels was found according to the presence of premature rupture of membrane, fetal tachycardia >160 bpm, meconial amniotic fluid, and duration of labour >10 h. No difference in sICAM-1 was noted between infected and non-infected infants. The cord serum levels of sICAM-1 were significantly higher in infants after forceps extraction (p = 0.01). A correlation was observed between sICAM-1 and C-reactive protein cord serum levels (p = 0.004, r = 0.371) and between sICAM-1 level and neutrophil count (p = 0.01, r = 0.489). CONCLUSIONS: Our results suggest that cord serum sICAM-1 determinations have no diagnostic relevance to neonatal infection. The increase of sICAM-1 cord serum levels in infants after forceps extraction suggests its potential to evaluate cerebral trauma or hypoxia. PMID- 11914571 TI - Prenatal diagnosis of fetal complete atrioventricular block with QT prolongation and alternating ventricular pacemakers using multi-channel magnetocardiography and current-arrow maps. AB - We report a case of fetal complete atrioventricular block (CAVB) with QT prolongation and alternating ventricular pacemakers diagnosed by magentocardiography (MCG). Fetal bradyarrhythmia of around 60 bpm was detected at 30 weeks of gestation. Ultrasonography revealed fetal CAVB without structural cardiac anomalies. MCG was recorded at 30 weeks of gestation using a 64-channel superconducting quantum interference device (MC-6400, Hitachi, Japan). The averaged MCG revealed QT prolongation, and the current-arrow maps generated by MCG revealed alternating ventricular pacemakers. The diagnosis was confirmed by electrocardiogram after birth. A permanent pacemaker was implanted during the early neonatal period, and the infant was healthy at 6 months of follow-up. MCG may be a useful diagnostic tool for fetal CAVB with QT prolongation and alternating ventricular pacemakers. PMID- 11914572 TI - Effect of OB/GYN residents' fatigue and training level on the accuracy of fetal weight estimation. AB - OBJECTIVE: To determine the effect of Ob/Gyn residents' fatigue and training level on the accuracy of their clinical and ultrasonographical estimation of fetal weight (EFW). METHODS: In this study, clinical and ultrasonographical EFWs were performed by various residents. Actual birth weight, gravidity, parity, gestational age, body mass index, presence or absence of diabetes and hypertensive diseases, presentation and amniotic fluid index were recorded. All EFWs were divided into 3 groups according to the hour they were performed. All residents were divided into 4 groups according to their training level. The accuracy of EFW as compared with actual birth weight was then analyzed according to the shift and to the residents' seniority by using the ANOVA test. Multivariate analysis was performed to evaluate the factors that significantly and independently affected the weight evaluation. RESULTS: Statistically significant differences were found between the clinical EFW and the birth weight among the working shifts for birth weights of 2,500 g and more (p = 0.032 and p = 0.035). For clinical EFW, night shifts were the most inaccurate (9.27, 8.05 and 9.78% of error for day, evening and night shift, respectively; p = 0.03). The accuracy of ultrasonographical EFW was not affected by the residents' fatigue level. The residents' training level did not alter the accuracy of either clinical or sonographical EFW. The accuracy of clinical EFW was affected independently by the work shift (p = 0.01), whereas no factor was found to independently effect the accuracy of ultrasonographic EFW. CONCLUSIONS: Ob/Gyn residents' fatigue affects the accuracy of clinical but not ultrasonographical EFWs. Residents' training level does not alter either the clinical or sonographical EFW. PMID- 11914573 TI - Alagille syndrome: prenatal diagnosis and pregnancy outcome. AB - The Alagille syndrome (AGS) is a multisystem autosomal dominant condition. In this case report, we describe a pregnant woman with this unusual disorder, in whom serial fetal sonography revealed severe pulmonary stenosis and progressively severe intrauterine growth retardation, suggesting that the fetus also had AGS, a diagnosis which was confirmed postnatally. In this report, the potential complications for pregnancy, labor and delivery when both mother and fetus are affected with AGS are described. PMID- 11914574 TI - Pathogenesis of acardiac twinning: clues from an almost acardiac twin. AB - OBJECTIVE: To search for clues to the pathogenesis of acardiac twinning. METHODS: We examined a case of monoamniotic twins in which twin A's only sonographic abnormality was a dilated, tortuous ductus venosus. Twin B also had this abnormality as well as multiple other anomalies that included enormous hydrops and a severely hypoplastic heart. Following termination of pregnancy, autopsy was performed. RESULTS: Postmortem examination of the placenta confirmed monochorionic, monoamniotic placentation with two adjacent trivascular cords. Autopsy confirmed the sonographic findings of enormous hydrops in twin B with a severely malformed, almost nonexistent heart. In addition, the liver was small and was represented by a cyst-like structure with thin rims of congested parenchyma surrounding large vascular spaces. CONCLUSION: We believe the sequence of events in this case was early twin-to-twin transfusion resulting in a dysfunctional heart in twin B. This enabled a twin reversal arterial perfusion sequence with further deterioration of twin B's heart and extreme congestion of deoxygenated blood exiting the heart into the inferior vena cava and ductus venosus. This case supports the concept that circulatory reversal in the face of an initially functioning heart may lead to congestion, tissue hypoxia and secondary organ atrophy. PMID- 11914575 TI - Linear insertion of atrioventricular valves without septal defect: a new anatomical landmark for Down's syndrome? AB - Our objective was to explore whether minor anatomical abnormalities of the septal insertion of tricuspid and mitral valves could be a feature of trisomy 21 in fetuses with an otherwise normal heart. Postmortem examinations were performed in 41 fetuses affected by Down's syndrome and in 52 controls. Adjoining the standard postmortem procedure, an apex-to-base section of the crux of the heart was made on a plane corresponding to the sonographic four-chamber view. This allowed gross and histological examination of the hinge points of tricuspid and mitral leaflets, showing the usual apical displacement of the tricuspid valve in all controls. Of 41 fetuses affected by Down's syndrome, 18 had a structural heart defect. Of the 23 Down syndrome fetuses without a patent heart defect, 16 (i.e., 69% of those considered as having 'normal hearts') had nevertheless a linear insertion of atrioventricular valves at autopsy. Prospective clinical studies are required to evaluate if these postmortem findings can be transposed to the clinical setting of 2nd-trimester sonographic screening. PMID- 11914576 TI - HIV vaccine development in the nonhuman primate model of AIDS. AB - Development of a prophylactic human immunodeficiency virus type 1 (HIV-1) vaccine is a leading priority in biomedical research. Much of this work has been done with the nonhuman primate model of AIDS. In a historical context, vaccine studies, which use this model, are summarized and discussed. PMID- 11914577 TI - Effects of prolactin-releasing peptide on tuberoinfundibular dopaminergic neuronal activity and prolactin secretion in estrogen-treated female rats. AB - Both systemic and central effects of a newly discovered prolactin (PRL)-releasing factor (PRF), prolactin-releasing peptide (PrRP), were determined in this study. Systemic injection of PrRP (1 and 10 microg/rat, i.v.) stimulated PRL secretion in ovariectomized, estrogen-treated rats similar to the effect of another PRF, thyrotropin-releasing hormone (TRH). Pretreatment with a dopamine D2 receptor antagonist, sulpiride (1 microg/rat, i.v.), potentiated the stimulatory effect of both PrRP and TRH on PRL secretion. Using the double-labeling immunohistochemical method, PrRP-immunoreactive terminals were found in close contact with tyrosine hydroxylase-immunoreactive neurons in the hypothalamic arcuate nucleus. Central administration of PrRP (0.1-1,000 ng/rat, i.c.v.) stimulated tuberoinfundibular but not nigrostriatal dopaminergic neuronal activity in 15 min. Levels of 3,4 dihydroxyphenylacetic acid (DOPAC) in the median eminence and striatum were used as indices for tuberoinfundibular dopaminergic (TIDA) and nigrostriatal dopaminergic neuronal activities, respectively. The serum PRL level, however, was not significantly changed. Similar treatment with TRH (10 ng/rat, i.c.v.) stimulated and inhibited TIDA neuronal activity and serum PRL, respectively, at 30 min. In summary, PrRP may play a role in both the central and peripheral control of PRL secretion. PMID- 11914578 TI - Tetrahydrobiopterin improves vascular endothelial function in ovariectomized rats. AB - The goal of the present study is to investigate the role of tetrahydrobiopterin (BH4) in the vascular response in ovariectomized rats. Rats were randomly assigned to two groups: (1) sham group: sham-operated female rats, and (2) Ovx group: rats were ovariectomized. Our results have shown that the plasma 17 beta estradiol levels in the Ovx group at the end of the experiment were significantly lower than in the sham group. Vasoreactivity assessed with intact aortic rings indicated that the phenylephrine-induced vasocontractile response to aortic rings from the Ovx group was greater than that of the sham group. In contrast, the vasodilator responses to acetylcholine and L-arginine (L-Arg) in the sham group were significantly greater than in the Ovx group. Differences in vasoreactivity in denuded aorta between the two groups were not noted. Moreover, exogenous BH4 significantly restored L-Arg-induced vasodilator responses in the Ovx group. However, this improvement effect was not found in the sham group. In addition, there were significant increases in superoxide anion production in aortic tissue and significant decreases in plasma nitric oxide levels in the Ovx group. Furthermore, BH4 contents in the aorta in the Ovx group were significantly decreased compared with the sham group. In conclusion, the present study demonstrates that the impairment of vascular reactivity was found in the ovariectomized rats. The possible mechanism of this defect may have resulted from the deficiency of available BH4. Thus, this study may provide a novel therapeutic strategy for the treatment of postmenopausal cardiovascular disorders. PMID- 11914579 TI - Co-administration of dopamine D1 and D2 agonists additively decreases daily food intake, body weight and hypothalamic neuropeptide Y level in rats. AB - This study investigated whether co-administration of dopamine D1 and D2 agonists might additively inhibit the feeding effect and whether this effect was mediated by the action on hypothalamic neuropeptide Y (NPY). The D1 agonist SKF 38393 (SKF) and D2 agonists apomorphine (APO) or quinpirole (QNP) were administered, alone or in combination, to examine this possibility. In single administration, decreases of daily food intake were observed only in rats treated twice a day with a higher dose of SKF, APO or QNP. However, combined administration of D1 and D2 agonists, with each agent at a dose that alone did not induce anorexia in one daily treatment, exerted a significant effect. These results reveal that co activation of D1 and D2 receptors can additively reduce daily food intake and body weight. The same treatment also decreased the level of hypothalamic NPY 24 h post-treatment. These results suggest an additive effect during combined activation of D1 and D2 receptor subtypes to decrease food intake and body weight that are mediated by the action of hypothalamic NPY. Similar to the effects seen in healthy rats, combined D1/D2 administration was also effective in the reduction of food intake in diabetic rats, revealing the efficiency of D1/D2 agonist in the improvement of hyperphasia in diabetic animals. PMID- 11914580 TI - Bcl-2 upregulation by HIV-1 Tat during infection of primary human macrophages in culture. AB - The ability of cells of the human monocyte/macrophage lineage to host HIV-1 replication while resisting cell death is believed to significantly contribute to their ability to serve as a reservoir for viral replication in the host. Although macrophages are generally resistant to apoptosis, interruption of anti-apoptotic pathways can render them susceptible to apoptosis. Here we report that HIV-1(BAL )infection of primary human monocyte-derived macrophages (MDM) upregulates the mRNA and protein levels of the anti-apoptic gene, Bcl-2. Furthermore, this upregulation can be quantitatively mimicked by treating MDM with soluble HIV-1 Tat-86 protein. These results suggest that in infecting cells of the monocyte/macrophage lineage, HIV-1 may be benefiting from additional protection against apoptosis caused by specific upregulation of cellular anti-apoptotic genes. PMID- 11914581 TI - A comparison of cholinesterase activity after intravenous, oral or dermal administration of methyl parathion. AB - Time-dependent changes in blood cholinesterase activity caused by single intravenous, oral or dermal administration of methyl parathion to adult female rats were defined. Intravenous and oral administration of 2.5 mg/kg methyl parathion resulted in rapid (<60 min) decreases in cholinesterase activity which recovered fully in vivo within 30-48 h. In contrast, spontaneous reactivation of cholinesterase in vitro was complete within 6 h at 37 degrees C. Dermal administration of methyl parathion caused dose-dependent inhibition of cholinesterase activity which developed slowly (> or =6 h) and was prolonged (> or =48 h). Time- and route-dependent effects of methyl parathion on cholinesterase activity in brain and other tissues generally paralleled its effects on activity in blood. In conclusion, pharmacodynamics of methyl parathion differ substantially with route of exposure. Recovery of cholinesterase in vivo after intravenous or oral exposure may partially reflect spontaneous reactivation and suggests a rapid clearance of methyl parathion or its active metabolite methyl paraoxon. The more gradual and prolonged inhibition of cholinesterase caused by dermal administration is consistent with disposition of methyl parathion at a site from which it or methyl paraoxon is only slowly distributed. Thus, dermal exposure to methyl parathion may pose the greatest risk for long term adverse effects. PMID- 11914582 TI - Chronic exercise increases both inducible and endothelial nitric oxide synthase gene expression in endothelial cells of rat aorta. AB - Chronic exercise upregulates endothelial nitric oxide synthase (eNOS) gene expression. Whether the expression of inducible nitric oxide synthase (iNOS) is affected by exercise is unknown. We therefore investigated the effects of chronic exercise on iNOS and eNOS expression in isolated rat aortic endothelial and smooth muscle cells separately. Five-week-old male Wistar rats were randomly divided into control and exercise groups. After 10 weeks of running training, animals were sacrificed under ether anesthesia. The standard curve quantitative competitive reverse transcriptase-polymerase chain reaction method was used to quantify NOS mRNA expression in isolated endothelial/smooth muscle cells. To evaluate the functional role of iNOS, we examined phenylephrine-induced vascular responses with or without pretreatment with aminoguanidine. We found that (1) expression of iNOS and eNOS mRNA in endothelial cells was increased by chronic exercise and (2) chronic exercise blunted phenylephrine-induced vascular responses, probably by increasing NO release via iNOS. Our results show that chronic exercise increases both iNOS and eNOS gene expression in endothelium. These alterations may be partially responsible for the change in vascular response after exercise. PMID- 11914583 TI - Functional role of extracellular signal-regulated kinase activation and c-Jun induction in phorbol ester-induced promoter activation of human 12(S) lipoxygenase gene. AB - The functional role of mitogen-activated protein kinase (MAPK) signaling and c Jun induction in phorbol 12-myristate 13-acetate (PMA)-induced human 12(S) lipoxygenase gene expression was studied in human epidermoid carcinoma A431 cells. Among the family of MAPK, PMA only increased the activity of extracellular signal-regulated kinase (ERK). Treatment of cells with PD98059, which is an inhibitor of mitogen-activated protein kinase kinase (MEK), decreased the PMA induced expression of 12(S)-lipoxygenase. Transfection of cells with Ras, Raf and ERK2 dominant negative mutants inhibited the PMA-induced promoter activation of the 12(S)-lipoxygenase gene in all cases. PMA-induced expression of c-Jun was inhibited by pretreatment with PD98059. Following treatment with PMA, the interaction between c-Jun and simian virus 40 promoter factor 1 (Sp1) in cells increased with time. Enhancement of binding between the c-Jun-Sp1 complex and the Sp1 oligonucleotide was observed in cells treated with PMA, suggesting the possible interaction of c-Jun-Sp1 with GC-rich binding sites in the gene promoter. These results indicate that PMA treatment induced ERK activation mainly through the Raf-MEK-ERK signaling pathway following induction of c-Jun expression, and the formation of the c-Jun-Sp1 complex. Finally, PMA activated the promoter activity of the 12(S)-lipoxygenase gene in cells overexpressing protein kinase C (PKC)delta but not PKCalpha, indicating that PKCdelta played the functional role in mediating the gene activation of 12(S)-lipoxygenase induced by PMA. PMID- 11914584 TI - Molecular epidemiology of hepatitis B viral serotypes and genotypes in taiwan. AB - Subtypes of hepatitis B virus (HBV) have specific geographic distributions and can serve as epidemiological markers. The relationship of HBV serotypes and genotypes in Taiwan and their correlation with the domiciles of origin in 122 patients with chronic HBV infection were investigated. The serotype of HBV was determined by comparing the surface gene encoding amino acids 22-148 of the major surface protein with published sequences. Genotyping of HBV was performed by polymerase chain reaction-restriction fragment length polymorphism. Serotype adw accounted for 70% (85/122) of all HBVs, with the remaining belonging to serotype adr. All adr HBVs were genotype C, regardless of the patient's domicile. Of the 85 adw HBVs, 69 (81%) were genotype B, 10 (12%) were genotype C, 5 (6%) were genotype F and only 1 (1%) was genotype A. In the 31 patients originating from mainland China, the prevalence of adr/genotype C was higher than in the 91 Taiwanese patients (15/31 vs. 22/91; p < 0.05). The distribution of the HBV serotypes and genotypes was not significantly different between 17 patients born in Taiwan (6 adw/genotype B, 2 adw/genotype C, 1 adw/genotype F and 8 adr/genotype C) and 14 patients born in mainland China (5 adw/genotype B, 2 adw/genotype C and 7 adr/genotype C). Our results indicate that in Taiwan, most HBVs of serotype adw are genotype B, and all HBVs of serotype adr are genotype C. Patients with origins in mainland China have a higher proportion of serotype adr/genotype C infection. PMID- 11914585 TI - Target integration by a chimeric Sp1 zinc finger domain-Moloney murine leukemia virus integrase in vivo. AB - A specificity protein 1 (Sp1) zinc finger domain containing two tandem zinc fingers was fused to the C terminus of the integrase (IN) protein of the Moloney murine leukemia virus (MuLV). The integrity of the MuLV IN was completely preserved, since the fusion was conducted at the last amino acid residue of the protein. The vector pMIN-Sp1, which carried the fused MuLV IN-Sp1 zinc finger domain gene, was cotransfected with a wild-type MuLV vector pMLV-K to NIH/3T3 cells. A nonradioactive reverse transcriptase assay was performed on culture supernatants collected from the cotransfected cells to confirm the production of recombinant viruses. The expression of the fusion protein and the integration of the MuLV genome by the fusion protein were confirmed by a Northern and then a Southern hybridization analysis on the total RNA or genomic DNA extracted from cells infected by viruses collected from the supernatants of the cotransfected cells. Regions of the host chromosome that were selected by the fusion protein as the integration targets were sequenced using the TOPO(TM) cloning method on a series of PCR products generated with a nested set of primers. The percentage of positive clones screened that contained the DNA-binding sequence of the fused Sp1 zinc finger domain was around 13% (5 out of 39 clones). It was found that the Sp1 DNA-binding sequence was only present in regions that were proximal to one of the long terminal repeats of the integrated viral genome, suggesting that the fusion protein could select a target sequence for integration. The host flanking sequences determined for all the positive clones were also used as queries to perform a BLAST search on the GenBank mouse EST entries. Although matching scores for sequences of some of the clones computed were more significant than others, it was difficult to judge whether or not the integration in these clones had been targeted to some gene sequences. Most of the integration sites might exist in the introns, since we found that the probability of the gene sequences containing an Sp1 DNA-binding site was low. PMID- 11914586 TI - Suppressive action of orexin A on pulsatile luteinizing hormone secretion is potentiated by a low dose of estrogen in ovariectomized rats. AB - Orexins are hypothalamic neuropeptides which stimulate luteinizing hormone (LH) secretion in estrogen- and progesterone-treated ovariectomized (OVX) rats and suppress it in OVX rats not treated with estrogen, suggesting a modulation by estrogen of the response to orexins. We examined the effects of orexin A on pulsatile LH secretion in OVX rats treated with a very small dose of estrogen so as to maintain the pulsatile secretion of LH. The estrogen treatment was done 24 h before the blood sampling by subcutaneously implanting a silicone tube (id = 1.5 mm, od = 2.5 mm, length = 25 mm) containing 17beta-estradiol (E(2)) dissolved in sesame oil at 20 microg/ml. In OVX rats treated with sesame oil as a control, the intracerebroventricular (icv) injection of orexin A (0.3 nmol, dissolved in 3 microl artificial cerebrospinal fluid) had no significant effect on the parameters of pulsatile LH secretion, i.e., pulse frequency and pulse amplitude, although it caused a small but statistically significant decrease in overall mean LH concentrations within 1 h. In OVX rats treated with E(2), the icv injection of orexin A significantly suppressed the pulsatile LH secretion; the frequency decreased for more than 2 h, inducing a rapid decline in overall mean LH concentrations. In view of the finding that a much higher dose of orexin A suppresses pulsatile LH secretion in OVX rats not treated with E(2), we suggest that the suppressive action of orexin A on pulsatile LH secretion is potentiated by estrogen. PMID- 11914587 TI - Timing of initiation of the preovulatory luteinizing hormone surge and its relationship with the circadian cortisol rhythm in the human. AB - The relationship between the hypothalamo-pituitary-gonadal (HPG) axis and the hypothalamo-pituitary-adrenal (HPA) axis has been well documented in the rat. In most cases, a negative coupling was observed and an inhibitory effect of the HPA axis upon the HPG was shown. In the female rat, a marked circadian rhythm of corticosterone plasma values is observed during each day of the estrous cycle, with maximal values around 08:00 p.m. The preovulatory luteinizing hormone (LH) surge also occurs at 08:00 p.m. on the day of proestrus. Here we measured circadian variations of plasma cortisol in humans in relation with the time of initiation of the preovulatory LH surge. Blood samples were taken at 08:00 a.m., 12:00 a.m., 04:00 p.m., 08:00 p.m., 12:00 p.m., and 04:00 a.m. from 19 subjects for 4 consecutive days, once 17beta-estradiol (E(2)) values reached 125 pg/ml (days 7-10 of the menstrual cycle). Serum E(2) and LH determinations were performed by microparticle enzyme immunoassays. Serum progesterone and plasma cortisol determinations were made using RIA methods. For plasma cortisol values, a marked circadian rhythm, with 2- to 3-fold higher values during the morning than during the afternoon, was almost identical before, during and after the LH surge. However, values were generally higher during the follicular phase than during the luteal phase. Maximum cortisol values occurred between 04:00 and 08:00 a.m. and minimal cortisol values between 04:00 and 08:00 p.m. Initiation of the LH surge (50% over the mean of previous values) occurred at 04:00 a.m. (20% of the cases) or at 08:00 a.m. (80% of the cases). There was a strong coupling between the onset of the surge and the acrophase of the cortisol circadian rhythm: maximal cortisol plasma values were seen at 04:00 a.m. when the LH preovulatory surge started at 04:00 a.m. and 08:00 a.m. when it started at 08:00 a.m. The present results show that the positive coupling documented in the female rat between the HPA and the HPG axis at the time of preovulatory LH surge is also present during the menstrual cycle in the human. PMID- 11914588 TI - Pituitary adenylate cyclase activating polypeptide and neuropeptide Y regulation of gonadotropin subunit gene expression in tilapia: role of PKC, PKA and ERK. AB - There is ample information on the hypophysiotropic function of pituitary adenylate cyclase-activating polypeptide (PACAP) and neuropeptide Y (NPY) in fish as in mammals, although evidence as to their direct effects on gonadotropic cells is scarce. We have previously reported that NPY and PACAP38 augment gonadotropin releasing hormone (GnRH)-induced expression of glycoprotein alpha (alpha) subunit gene in the teleost fish, tilapia. The aim of the present study was to elucidate possible direct effects of these peptides on gonadotropin subunit gene expression in culture of tilapia pituitary cells, as well as the transduction pathways involved. Both NPY and PACAP38 (0.001-10 nM) increased the level of phosphorylated extracellular signal-regulated kinase (pERK) dose-dependently, reaching a peak at 0.1 and 0.01 nM, respectively. Inhibition of protein kinase C (PKC) by GF109203X (GF; 0.01-10 nM) suppressed NPY-stimulated pERK levels and its effect on alpha and luteinizing hormone (LH) beta subunit mRNA levels. However, NPY had no effect on follicle stimulating hormone (FSH) beta mRNA levels. NPY elevated alpha, LHbeta mRNA and pERK levels were also attenuated by inhibition of protein kinase A (PKA) with H89 (0.01-10 nM). Exposure of the cells to the MAPK kinase (MEK) inhibitor (PD98059; PD 10, 25 and 50 microM) completely blocked NPY induced ERK activity. In addition, this inhibitor abated the alpha and LHbeta mRNA responses to NPY. Similar experiments conducted to elucidate PACAP38 signaling revealed that PACAP38 (0.01 nM) elevated all three-gonadotropin subunit gene expression via both PKC-ERK and PKA-ERK cascades. It is suggested that both NPY and PACAP38 act directly on gonadotropes to elevate gonadotropin subunit gene expression. Whereas the expression of alpha and LHbeta subunit genes is regulated by both NPY and PACAP, the effect on the FSHbeta transcript is elicited only by PACAP38. NPY and PACAP38 stimulatory actions are mediated via protein kinase C (PKC) and protein kinase A (PKA), converging at the MEK-ERK cascade. These findings represent one of the fine tuning levels that differentially regulates gonadotropin subunit gene expression. PMID- 11914589 TI - Infusion of alpha-2-adrenergic agents into the paraventricular and arcuate nuclei of the hypothalamus in the Siberian hamster: opposing effects on basal prolactin. AB - Although the alpha(2)-adrenergic receptor subtype has been shown to have a significant influence on circulating levels of prolactin (PRL), its exact role remains unclear. A multitude of studies have demonstrated that blockade of the alpha(2)-receptor can either elevate or decrease circulating levels of PRL. Alpha(2)-receptor-mediated control of both stimulatory and inhibitory arms of the PRL regulatory system may explain this discrepancy. Activation of the alpha(2) receptor has been shown to inhibit the activity of its target cell, and therefore antagonism of the alpha(2)-receptor within a stimulatory component (e.g., paraventricular nucleus (PVN) of the hypothalamus) would theoretically have the opposite effect that it would have within an inhibitory component (arcuate nucleus of the hypothalamus). Thus, the purpose of this study was to investigate the functional role of the alpha(2)-adrenergic receptor in modulating circulating levels of PRL both at the level of the PVN and arcuate using reverse microdialysis of alpha(2)-adrenergic agents coupled with serial blood sampling in the male Siberian hamster. Male hamsters were fitted with a jugular cannula for serial blood sampling, and an indwelling microdialysis probe for intrahypothalamic drug administration between 08:00 and 10:00 h. Blood samples were collected every hour for 5 h (12:00-17:00 h). During the third sampling period, atipamezole (alpha(2)-antagonist) or medetomidine (alpha(2)-agonist) at one of three doses were infused into the PVN or the arcuate to assess effects on basal PRL. At the level of the PVN, infusion of atipamezole initiated an increase in basal PRL in a dose-dependent fashion, whereas infusion of medetomidine induced a significant decline in basal PRL in a dose-dependent fashion. In the arcuate, only the highest dose of atipamezole had an effect on PRL, and this was in the opposite direction from that seen in the PVN. Infusion of medetomidine did not have a significant effect on basal PRL levels; however, a trend toward a significant elevation was observed for the highest dose. These results suggest that the alpha(2)-receptor subtype may have opposite effects on circulating levels of PRL within the PVN and arcuate regions, and may explain why antagonism of the alpha(2)-receptor has been shown to initiate both surges and declines in basal levels of PRL. PMID- 11914590 TI - Regulation of prolactin secretion by dopamine at the level of the hypothalamus in the turkey. AB - Avian prolactin (PRL) secretion is regulated by vasoactive intestinal peptide (VIP) neurons residing in the infundibular nuclear complex (INF) of the hypothalamus. This VIPergic activity is modulated by stimulatory dopaminergic inputs. Dynorphin, serotonin (5-HT), dopamine (DA) and VIP all appear to stimulate PRL secretion along a hypothalamic pathway, expressing kappa opioid, serotonergic, dopaminergic and VIPergic receptors in succession, with the VIPergic system as the final mediator. Electrical stimulation (ES) within the turkey hypothalamus at the level of the medial preoptic area (POA), the ventromedial hypothalamic nucleus (VMN), the INF or the median eminence (ME) results in the release of PRL. When the selective D(1) DA receptor antagonist SCH 23390 HCl was infused intraventricularly at the rate of 10 nmol/min, ES in the POA or VMN was unable to increase PRL levels, while ES in the INF and ME did increase PRL to the same level as that of controls. These results were interpreted to suggest that the D(1) DA receptors involved in PRL release lie caudally to the VMN and dorsally to the INF. Bilateral microinjections (50 ng) of the D(1) DA receptor agonist SKF-38393 HCl into the POA or VMN failed to produce any increase in PRL, while similar microinjections in the INF increased PRL significantly within 15 min. Bilateral microinjections of the D(1) DA antagonist (50 ng) into the INF blocked the rise in PRL associated with ES in the POA. Bilateral microinjections of a D(2) DA antagonist (50 ng) into the INF failed to block PRL secretion induced by ES in the POA. Tract tracing, using double-label immunocytochemistry, revealed the presence of a monosynaptic dopaminergic pathway projecting from the POA to the INF. These data imply that the only hypothalamic D(1) DA receptors involved in the regulation of avian PRL secretion are those residing within the INF in the same region as the VIP neurons known to be involved in PRL secretion. PMID- 11914591 TI - Estradiol stimulates gene expression of norepinephrine biosynthetic enzymes in rat locus coeruleus. AB - Gender-specific differences in susceptibility to a number of disorders related to catecholaminergic systems, including depression and hypertension, have been postulated to be mediated, at least in part, by estrogens. In this study, we examined if estrogens may regulate gene expression of norepinephrine biosynthetic enzymes. Administration of five injections of 15 or 40 microg/kg estradiol benzoate to ovariectomized (OVX) female rats elicited a dose-dependent elevation in mRNA levels of tyrosine hydroxylase (TH) in locus coeruleus, to as great as 3 fold over control. Dopamine beta-hydroxylase (DBH) mRNA levels were also similarly increased. To examine the mechanism, PC12 cells were cotransfected with luciferase reporter constructs under control of DBH or TH promoters [pDBH/Luc( 2,236/+21) or pTH/Luc(-272/+27 or -773/+27)] with an expression vector for estradiol receptor alpha. The cells were treated with 17beta-estradiol (E(2)) for 12-36 h. E(2) triggered a several fold increase in luciferase activity under control of the DBH promoter in a dose-dependent fashion. Omission of estrogen receptor alpha or addition of the estrogen receptor antagonist ICI 182,780 prevented the DBH promoter-driven increase in luciferase. When E(2) was given with 0.2 mM CPT-cAMP, reporter activity with pDBH/Luc(-2,236/+21) was increased greater than with either treatment alone. In contrast, addition of E(2) to cells transfected with pTH/Luc(-272/+27) elicited no change in basal luciferase activity nor in the response to 0.2 mM CPT-cAMP. These findings are the first to reveal that estrogen can stimulate DBH gene expression. Differing mechanisms may underlie the regulation of TH and DBH gene expression by estrogens. PMID- 11914592 TI - Circadian rhythms of dopamine and dihydroxyphenyl acetic acid in the mouse striatum: effects of pinealectomy and of melatonin treatment. AB - The existence of dopamine (DA)-melatonin (aMT) relationships is well documented in several brain areas of the mammalian central nervous system such as the retina and hypothalamus or the nigrostriatal system. For instance, aMT tempers 1 methyl 4-phenyl-1,2,3,6-tetrahydropyridine-induced nigrostriatal damage in C57BL/6 mice. In this mouse strain however, rhythmic production of aMT and its possible interaction with striatal DA is still unclear. In the present work we investigated circadian variations in pineal production of aMT and striatal DA levels in C57BL/6 mice. Effects of pinealectomy and aMT administration were also assessed. Intact, pinealectomized and pinealectomized + aMT-treated mice and their respective control groups were sacrificed at six different times during the 24-hour period. In control animals, aMT displayed a circadian rhythm with a narrow peak at midnight. The peak of aMT coincided with the nadir of the DA rhythm present in the striatum. Shortly after the decrease of DA levels, an increase in 3,4-dihydroxyphenylacetic acid (DOPAC), the main DA metabolite, was observed. The rhythmic changes of DA and DOPAC levels in the striatum were blunted by pinealectomy, whereas administration of aMT (0.1-10 mg/kg) during 6 days to pinealectomized mice restored the rhythms in a dose-dependent manner. Striatal levels of 3-methoxytyramine and homovanillic acid did not change during the 24-hour cycle. The serotonergic system, assessed by the determination of 5 hydroxytryptamine and 5-hydroxyindole-3-acetic acid concentration in striatum, did not show significant time-dependent changes in control animals and was not affected by pinealectomy or aMT treatment. These data substantiate the existence of a link between pineal function, melatonin secretion and DA circadian rhythm in the mouse striatum. PMID- 11914593 TI - Effectiveness of the MSC cold cap system in the prevention of chemotherapy induced alopecia. AB - OBJECTIVE: To study the effectiveness of the MSC cold cap system to prevent chemotherapy-induced alopecia. METHODS: The system was applied in 83 cancer patients (mean age 49.8 years) undergoing chemotherapy with alopecia-causing agents. Seven patients did not tolerate the system. Seventy-six patients were evaluable for assessment; 26 received anthracycline (group A), 33 taxane (group T), 5 anthracycline plus taxane (group AT), 7 intravenous etoposide (group E) and 5 ifosfamide with or without other alopecia-causing drugs (group I). In group A, 18 patients received conventional (subgroup Ac) and 8 high doses (subgroup Ah). In group T, 8 patients received docetaxel (subgroup D) and 25 paclitaxel (subgroup P). Alopecia grade 0-1 (Dean's system) was considered as treatment success. RESULTS: Grade 0-1 alopecia was achieved in 49/76 (64.5%) patients: group T 23/33 (69.6%), subgroup P 16/25 (64%) and subgroup D 7/8 (87.5%); group A 18/26 (69.2%), subgroup Ac 16/18 (88.8%) and subgroup Ah 2/8 (25%); group AT 1/5 (20%); group E 6/7 (85.7%), and group I 1/5 (20%). CONCLUSIONS: The MSC cold cap system is effective in preventing alopecia from anthracycline, etoposide or taxane but not from anthracycline-taxane combinations or ifosfamide-containing regimens. PMID- 11914594 TI - Phase I study of vinorelbine and carboplatin combination in patients with taxane and anthracycline pretreated advanced breast cancer. AB - AIM: To define the maximum tolerated dose (MTD) and the dose-limiting toxicities (DLTs) of the carboplatin-vinorelbine combination in pretreated patients with advanced breast cancer. PATIENTS AND METHODS: Patients with histologically confirmed metastatic breast cancer relapsing or progressing after prior taxane and anthracycline containing chemotherapy were enrolled. Cohorts of 3-6 patients were treated at successive dose levels (DLs) with escalated doses of carboplatin [range, area under the curve (AUC) 4-6] on day 1 and vinorelbine (range, 20-35 mg/m(2)) on days 1 + 8 recycled every 28 days. RESULTS: Twenty-seven patients with a median age of 58 years and performance status (WHO) of 0-2 were treated at 6 DLs. All patients were assessable for toxicity and 20 for response. DLT was reached at carboplatin 6 AUC and vinorelbine 35 mg/m(2), and therefore, this was considered as the MTD. Prophylactic G-CSF administration could not allow further dose escalation. The recommended dose for further phase II testing was defined at carboplatin 6 AUC on day 1 and vinorelbine 30 mg/m(2) on days 1 and 8. Among 98 administered treatment cycles 41 (42%) and 7 (7%) were complicated with grades 3 and 4 neutropenia and thrombocytopenia, respectively. Nonhematologic toxicities included grade 2 peripheral neuropathy in 3 cycles and grades 2 and 3 fatigue in 32 (32%). CONCLUSION: The present study determined the feasibility of the combination of carboplatin at AUC 6 (day 1) and vinorelbine at 30 mg/m(2) (days 1 and 8 ) without G-CSF support in patients with taxane and anthracycline pretreated advanced breast cancer. Phase II studies at these doses should follow in order to determine the activity of the regimen. PMID- 11914595 TI - Phase I study of gemcitabine and liposomal doxorubicin in relapsed ovarian cancer. AB - Twenty-three patients were enrolled in a phase I study conducted to determine the maximum tolerated doses (MTD) of combined liposomal doxorubicin (CAE) and gemcitabine (GEM) in relapsed ovarian cancer patients. A total of 82 courses are evaluable, with a median number of three cycles administered per patient (range 2 8). GEM was administered on days 1 and 8 by 30-min intravenous infusion immediately after CAE given by 60-min intravenous infusion on day 1; cycles were repeated every 21 days. The starting doses were CAE 20 mg/m(2) and GEM 600 mg/m(2). Following dose levels were 20/800; 20/1,000; 30/800; 30/1,000; 35/800, and 35/1,000 for CAE and GEM, respectively. The MTD was reached at dose level 5, with febrile neutropenia and thrombocytopenia as dose-limiting toxicities. After the MTD, granulocyte-colony stimulating factor was administered in 15% of cycles. Non-hematological toxicity was mild and manageable. All patients are so far evaluable for response. Among them, 5 partial responses (21.7%; 95% confidence interval, CI: 4.9-38.5), 5 disease stabilizations (21.7%, 95% CI: 4.9-38.5) and 13 progressions (56.6%, 95% CI: 36.4-76.8) have been registered. These results warrant further research in a phase II study. PMID- 11914596 TI - Are the effects of tamoxifen on the serum lipid profile modified by apolipoprotein E phenotypes? AB - BACKGROUND: Tamoxifen has favorable effects on the serum lipid profile. It has been suggested that the apolipoprotein (Apo) E phenotype can influence serum lipid parameters; the ApoE allele 4 (ApoE4) is associated with higher total and low-density lipoprotein (LDL) cholesterol levels. The ApoE phenotype also affects lipid responses to diets or treatment with statins. However, the effect of tamoxifen on the lipid profile in different ApoE phenotypes is unknown. PATIENTS AND METHODS: In the present study, we evaluated the effects of tamoxifen on the serum lipid profile in 11 ApoE4-positive postmenopausal women with breast cancer (phenotypes 3/4 and 4/4) compared with 33 ApoE4-negative women (phenotypes 3/2 and 3/3). Serum lipid parameters [high-density (HDL), LDL and total cholesterol, triglycerides, ApoAI, ApoB and lipoprotein (a)] were measured after an overnight fast before treatment and after 3 and 12 months. ApoE isoforms were determined by isoelectric focusing of delipidated very-low-density lipoproteins (VLDL). RESULTS: During the follow-up period, serum levels of total and LDL cholesterol and ApoB decreased significantly in both groups, but no significant differences were found. Concentrations of serum HDL cholesterol were not significantly different between both groups. However, serum ApoAI levels increased significantly in ApoE4-negative subjects (p = 0.00005), but no significant changes in ApoE4-positive women were observed. Serum triglyceride levels increased by 23.2% (p < 0.05) in ApoE4-positive patients, but they did not change significantly in ApoE4-negative patients. The LDL/HDL cholesterol ratio decreased similarly in the two groups, but the ApoAI/ApoB ratio, which may be a better predictor of cardiovascular events, significantly changed in the ApoE4-negative subjects. Finally, the median level of Lp(a) decreased by 43.4% in the ApoE4 negative patients, whereas it did not change significantly in the ApoE4-positive group. CONCLUSION: In postmenopausal Greek women with breast cancer, the levels of Lp(a) and triglycerides and the ApoAI/ApoB ratio respond more favorably to tamoxifen treatment in ApoE4-negative than in ApoE4-positive patients. PMID- 11914598 TI - Flow-cytometric determination of tumor cells in lymph nodes. AB - In solid tumors, metastasis occurs through the dissemination of tumor cells in the bloodstream and the lymphatic system. In particular, lymph node infiltration gives useful prognostic information and represents one of the most important factors for selecting the type of clinical treatment in disease management. Furthermore, the analysis of lymph node infiltration has become important for identifying patients with breast cancer or malignant melanoma who may be candidates for regional lymph node dissection. Tumor cells in lymph nodes are currently identified in tissue sections using morphological and immunohistochemical analyses, but these approaches are time-consuming, and micrometastases may escape detection. The aim of the present study was to define the potential of a flow cytometric (FCM) determination based on cell size and autofluorescence to shorten the time required for lymph node analysis. The sensitivity of the FCM approach, defined on mixtures of tumor cells from established cell lines and peripheral blood lymphocytes (PBL(s)) at different concentrations, was 1 tumor cell/1,000 PBL(s). FCM analysis was performed on 89 lymph nodes, 29 from breast, 41 from lung and 19 from colon cancer patients. Agreement between FCM and morphological results, used as gold standard, was observed in 83% of the cases, and there was a 90% sensitivity to the FCM approach for each tumor type. Disagreement was observed for 15 lymph nodes and was due, in the majority of cases (80%), to FCM-positive and morphologically negative results. A large number of patients and a more accurate pathological examination of consecutive histological sections of lymph nodes are needed to further evaluate the validity of the FCM approach. PMID- 11914597 TI - Correlation of vascular endothelial growth factor-C expression with tumor infiltrating dendritic cells in gastric cancer. AB - It has been reported that dendritic cells (DCs) play a crucial role in the host's immune defense against tumors, and there is an inverse correlation between the density of DCs and the expression of vascular endothelial growth factor (VEGF). However, the relationship between the expression of VEGF-C and DC infiltration remained unclear. In the present study, we investigated whether the expression of VEGF-C correlated with the number of DCs in vivo. We immunohistochemically analyzed gastric carcinoma tissue in this study. The survival curves show that the prognosis for patients with a low density of DCs was significantly poorer than that for patients with high DC density (p < 0.01). Further, the survival curves according to the VEGF-C status showed that the survival rate in patients with low VEGF-C expression was higher than that in patients with higher expression (p < 0.01). There was a significant negative correlation between the density of DCs and the expression of VEGF-C (r = -0.26, p < 0.05). We suggest that VEGF-C produced by cancer causes DCs to become dysfunctional. This may be one of the ways that cancers evade immunosurveillance. PMID- 11914599 TI - ARHI/NOEY2 inactivation may be important in breast tumor pathogenesis. AB - Allelic loss frequently occurs on the short arm of chromosome 1 in human breast carcinoma, suggesting that the ARHI/NOEY2 gene, an imprinted putative tumor suppressor gene, is involved in the pathogenesis of the tumor entity. To clarify the clinical importance of ARHI/NOEY2 mRNA in breast cancer, we studied whether ARHI/NOEY2 inactivation might contribute to tumors arising in the breast. An ARHI/NOEY2 message was detected by real-time PCR analysis in all noncancerous breast tissues, but was not detected in 2 of 26 breast cancer tissue samples. In 10 of 26 breast cancer tissue samples ARHI/NOEY2 mRNA was substantially reduced. ARHI/NOEY2 expression was lost or markedly reduced in 12 of 26 (46.15%) breast cancer tissue samples. In summary, we conclude that decreased ARHI/NOEY2 mRNA expression may play an important role in the pathogenesis of breast cancer. PMID- 11914600 TI - Prognostic impact of multiple allelic losses on metastatic recurrence in hepatocellular carcinoma after curative resection. AB - Loss of heterozygosity (LOH) on chromosomes 13q, 16q and 17p has been associated with the progression of hepatocellular carcinoma (HCC). To investigate the prognostic impact of such LOH, we examined the metastasis-free survival of curatively resected HCC cases, in whom these LOHs were analyzed. Among the 49 HCC patients examined, the frequency of LOHs was 28% on 13q, 33% on 16q and 40% on 17p. The patients were followed up for metastatic recurrence after surgery and for analysis of the relationship between chromosomal changes and patients' metastasis-free survival. Univariate survival analysis showed the presence of LOH on 16q, 17p and the number of chromosomes with LOH were significantly and negatively associated with metastasis-free survival, indicating that patients with LOH on multiple chromosomes had a poorer prognosis after surgery than those with LOH on a single chromosome or no LOH. Multivariate Cox survival analysis identified the presence of LOH on 16q and the number of chromosomes with LOH as the most significant independent negatively predictive factors for metastasis free survival. These findings indicate that accumulation of chromosomal changes is associated with metastatic behavior, and that LOH on 16q was the most useful prognostic indicator for metastasis after curative resection of HCC. PMID- 11914601 TI - Overexpression of beta 1,4N-acetylgalactosaminyl- transferase mRNA as a molecular marker for various types of cancers. AB - OBJECTIVE: To determine GalNAcT mRNA expression in human carcinoma cell lines and primary tumor tissues. Assessment of the potential use of GalNAcT mRNA as a molecular marker for detection of metastatic cancer cells in the peripheral blood of patients with hepatocellular carcinoma. METHODS/RESULTS: We investigated GalNAcT mRNA expression in various human cancer cell lines and primary cancer tissues using RT-PCR assay for GalNAcT mRNA. The expression of GalNAcT mRNA was detected in 25 of 26 cancer cell lines tested and in the majority of primary tumors from different organs: 8 of 10 colon cancers, 9 of 9 breast cancers, 11 of 12 esophageal cancers, 14 of 14 gastric cancers, 4 of 18 pancreatic cancers, 6 of 12 biliary tract cancers, 17 of 18 hepatocellular carcinomas and 13 of 14 lung cancers. Semi-quantitative analysis with duplex RT-PCR showed that the amount of the GalNAcT mRNA was enhanced in cancer tissues as compared to the surrounding cancer-free tissues. Blood specimens of 5 of 14 patients with hepatocellular carcinoma were positive for GalNAcT mRNA, all of whom developed recurrent disease in less than 24 months. Peripheral blood samples of 30 normal subjects were negative for GalNAcT mRNA. CONCLUSION: Our results suggest that the RT-PCR assay for GalNAcT mRNA could be a potentially useful molecular marker for detecting cancer dissemination in blood circulation of patients with malignancy. PMID- 11914602 TI - Clinical significance of vascular endothelial growth factor C expression and angiogenesis at the deepest invasive site of advanced colorectal carcinoma. AB - AIM: Vascular endothelial growth factor C (VEGF-C) is known to be associated with the development of the lymphatic vascular system. The aim of this study was to elucidate the clinical significance of VEGF-C expression and microvessel density (MVD) at the deepest invasive site in advanced colorectal carcinoma (CRC). METHODS: 152 patients who had undergone surgical resection for advanced CRC entered this study. VEGF-C expression was examined immunohistochemically with anti-VEGF-C polyclonal antibody C-20. Tumor MVD was determined immunohistochemically with anti-CD34 antibody. VEGF-C expression was defined as positive if distinct staining of the cytoplasm was observed in at least 10% of tumor cells at the deepest invasive site, central portion and superficial part of the tumor. MVD was estimated by averaging the count of three x400 fields in the most vascular area at the deepest invasive site. RESULTS: VEGF-C expression was detected in 71 of 152 lesions (46.7%) at the deepest invasive site. VEGF-C expression correlated significantly with poorer histologic grade, depth of invasion, lymphatic invasion, lymph node metastasis, venous invasion, liver metastasis and Duke's stage. At the central portion and superficial part, there were no significant differences between VEGF-C expression and clinicopathological findings. VEGF-C expression at the deepest invasive site also correlated significantly with MVD. In cases with curative surgery, patients with VEGF-C expression at the deepest invasive site had a significantly poorer prognosis than those without VEGF-C expression. Furthermore, prognosis for patients with both VEGF-C expression and high MVD at the deepest invasive site was significantly poorer than that of patients without VEGF-C expression and with low MVD. Multivariate analysis with logistic regression for 5-year survival in patients with curative surgery showed that lymph node metastasis and VEGF-C expression were significant risk factors. CONCLUSION: VEGF-C expression at the deepest site of tumor invasion can be a useful predictor of poor prognosis in advanced CRC and show a close relation to angiogenesis. PMID- 11914604 TI - p53 protein accumulation in multiple oesophageal squamous cell carcinoma: relationship to risk factors. AB - To clarify mechanisms involved in the carcinogenesis of multiple oesophageal squamous cell carcinoma, the expression of p53 protein in 46 lesions surgically excised from 13 Japanese patients was investigated immunohistochemically and the relation of p53 protein accumulation to the patient's history of alcohol consumption and cigarette smoking was analyzed. p53 protein accumulation was observed in 13 main lesions, that is in 6 (85.7%) of 7 subjects with a history of heavy drinking and smoking, but only in 1 (16.7%) of 6 with no such history (Fisher's exact test, p = 0.025). As regards the 46 lesions, p53 protein accumulation was evident in 22 (88.0%) of 25 lesions of the high-risk patients, but in 7 (33.3%) of 21 lesions of the other subjects (Fisher's exact test, p < 0.001). p53 protein accumulation was similarly recognized in all oesophageal lesions in 5 of 7 high-risk patients. Thus, use of both alcohol and cigarettes is clearly associated with a high frequency of p53 protein accumulation in multiple oesophageal squamous cell carcinoma present at the same time. These findings are considered to support the concept of field carcinogenesis of the oesophagus. PMID- 11914603 TI - Prognostic significance of the thymidylate biosynthetic enzymes in human colorectal tumors. AB - The aim of the present study was to determine the relative intratumoral activity of two pyrimidine biosynthetic enzymes, i.e. thymidylate synthase (TS) and thymidine kinase (TK), in human colorectal cancers to compare their possible relationship with demographic and pathologic characteristics of the patients and their tumors, and moreover to evaluate their predictive significance regarding 5 fluorouracil (5-FU) sensitivity and the overall survival of patients, respectively. TS and TK levels were significantly increased in the tumor compared to peritumoral tissue. However, no significant relationship between TS/TK activity and demographic features of the patients or pathologic characteristics of their tumors could be demonstrated. Kaplan-Meier analysis showed that the overall survival of patients with low TS activity was significantly longer (p < 0.012) compared to those with high TS activity. Such a difference could not be demonstrated between patients with high or low TK activity; however, combined evaluation of the two parameters proved that TK may contribute to the more precise assessment of disease prognosis, and it may further influence treatment decisions, i.e. the selection of patients for adjuvant therapy with 5-FU and folinic acid. Multivariate analysis showed that among the variables tested, beside Dukes' stage, TS and TK activities were significant prognostic factors for the overall survival of colorectal cancer patients. PMID- 11914605 TI - Establishment of a new cell line (TAYA) of clear cell adenocarcinoma of the ovary and its radiosensitivity. AB - A new cell line (TAYA) was established from ascites of a patient with clear cell adenocarcinoma of the ovary, and the sensitivity of the cell line to various anticancer drugs and radiation was investigated. The 50% growth inhibitory concentrations (IC50) (nM) of various anticancer drugs were: cisplatin >10,000, paclitaxel 80, SN-38 (irinotecan metabolite) 460, and gemcitabine >10,000, showing that the cells had a low sensitivity to these anticancer drugs. In contrast, the 50% growth inhibitory dose (ID50) of radiation was 1.8 Gy, showing that the cells were highly sensitive to radiation. Thus, radiotherapy may be effective for treatment of clear cell adenocarcinoma of the ovary. PMID- 11914606 TI - Choroidal vasculature changes in spontaneously hypertensive rats - transmission electron microscopy and scanning electron microscopy with casts. AB - PURPOSE: The purpose of the present study was to elucidate the specific morphological changes of the choroidal vasculature in long-term hypertension. METHODS: The choroidal vessels of spontaneously hypertensive (SHR) rats were examined with corrosion casts/scanning electron microscope (SEM) and the transmission electron microscope (TEM). RESULTS: In 18-month-old SHRs, corrosion casts and SEM showed tortuosity, caliber irregularity and generalized narrowing of the choroidal arteries. The draining venules were prominently decreased in number. The ampullae of the vortex veins were narrow, but arteriovenous anastomoses, crossing defects and obstruction were not seen. The choriocapillaris was elongated and engorged. TEM of the choroidal vessels showed hypertrophy of smooth muscle cells and irregular thickening of the basement membrane and narrow lumen of the choriocapillaris. The RPE and Bruch's membrane were intact. CONCLUSION: The choroidal vascular bed showed extensive morphological changes. This study revealed that hypertensive choroidopathy has specific features such as tortuosity, caliber irregularity, generalized narrowing of arteries, decreased number of draining venules and elongated and engorged choriocapillaris. These findings indicate that hypertensive choroidopathy is as important as retinopathy. PMID- 11914607 TI - Mucin-like glycoprotein associated with photoreceptor cells in the postnatal developing rat retina. AB - PURPOSE: To examine the developmental expression of mucin-like glycoprotein associated with photoreceptor cells (MLGAPC), and to confirm that MLGAPC with a relative mass of 120 and 135 kD correspond to the cone and rod types, respectively. METHODS: We isolated the cDNA of the core protein of rat MLGAPC, prepared an anti-rat MLGAPC core protein antibody and then examined the expression of the MLGAPC core protein in postnatal developing rat retinas by means of Western blotting and immunohistochemistry. RESULTS: On postnatal early days when cone photoreceptor cells were thought to be developing, MLGAPC was detected as a single band of 120 kD and was found to be positive in the apical portion of the neuroblastic layer. After postnatal day 7, when rod photoreceptor cells were developing, MLGAPC was detected as a minor band of 120 kD and an additional major band of 135 kD and was present throughout the differentiated photoreceptor and outer plexiform layers. CONCLUSIONS: The present findings revealed the developmental expression of MLGAPC and confirmed that MLGAPC of 120 and of 135 kD correspond to the cone and rod types, respectively. This demonstration is advantageous for studying the physiological roles of MLGAPC in the photoreceptor and outer plexiform layers, since the cone- and rod-associated glycoproteins should be investigated separately. PMID- 11914608 TI - Further characterization of human mucin-like glycoprotein associated with photoreceptor cells by its introduction into Y79 retinoblastoma cells. AB - PURPOSE: To understand the function of a mucin- like glycoprotein associated with photoreceptor cells (MLGAPC), MLGAPC-transfected Y79 retinoblastoma cells were analyzed biochemically and morphologically. METHODS: The cDNA for the core protein of human MLGAPC was isolated, cloned into a mammalian expression vector, pCAGGSneo, and then transfected into Y79 retinoblastoma cells. The cells and the medium were analyzed by means of Western blotting, and the morphology of the transfectants and parental cells was compared. RESULTS: Western blot analysis of the culture medium revealed that the transfectants secreted MLGAPC into the medium. Lectin blot analysis of MLGAPC in the medium showed that it had a binding site for Maackia amurensis lectin II. No morphological difference was detected between the transfectants and parental cells. CONCLUSIONS: As expected from the deduced amino acid sequence of MLGAPC, it was secreted into the medium. The secreted MLGAPC was found to carry sialoglycans (rod type). The expression of MLGAPC had no effect on the adhesion or morphology of the cultured cells, which suggests that its interaction with other components may be required for these effects. PMID- 11914609 TI - Apoptosis of human retina and retinal pigment cells induced by human cytomegalovirus infection. AB - Human cytomegalovirus (HCMV) retinitis is the most common ocular opportunistic infection in immunocompromised patients and AIDS. It often leads to blindness if left untreated. The question as to how HCMV infection causes retinal pathogenesis and visual destruction in AIDS patients remains unresolved. To answer the question, by using terminal deoxynucleotidyl transferase-mediated deoxyuridine triphosphate nick end labeling assay, we detected the significant signals of apoptotic cells at the same sites in the HCMV-infected retina of AIDS patients as compared to AIDS patients without HCMV retinitis. In vitro study also revealed apoptosis induced by HCMV infection in human retinal pigment epithelium cells, mediated by activation of caspase 3 and poly(ADP-ribose) polymerase pathway. These results strongly suggest the fundamental role of HCMV-induced apoptosis in mediating cell death in infected human retina and retinal pigment epithelium cells to make severe visual impairment. PMID- 11914610 TI - Multifocal electroretinogram with a multiflash stimulation technique in open angle glaucoma. AB - PURPOSE: To analyze a multiflash multifocal electroretinogram in 20 patients with open-angle glaucoma (OAG). METHODS: The stimulation sequence consisted of a binary m-sequence (L(max) 200 cd/m(2), L(min) <1 cd/m(2)). Each m-sequence stimulus was followed by three global flashes (luminance: 400 cd/m(2)) at an interval of 26 ms. RESULTS: The presence of a response to the three global flashes indicated an adaptive effect of the response to the preceding m-sequence stimulus. In the nasal retinal response average the relative amplitude contribution of the response to the second global flash in relation to the other two global flash responses was outside the range of normal (10th-90th percentile) in 10 of 20 OAG patients. CONCLUSIONS: The changes in the relative contribution of the response to the second global flash seem indicative of impaired adaptive effects presumably due to inner retinal damage. PMID- 11914611 TI - Effects of isopropyl unoprostone, latanoprost, and prostaglandin E(2) on acute rise of aqueous flare in pigmented rabbits. AB - PURPOSE: To evaluate the effect of isopropyl unoprostone, latanoprost, and prostaglandin E(2) (PGE(2)) on aqueous flare elevation. METHODS: Isopropyl unoprostone (0.12%) or latanoprost (0.005%) was topically instilled. Transcorneal diffusion of PGE(2), 25 microg/ml, using a glass cylinder, was achieved in pigmented rabbits. Aqueous flare was measured with a laser flare cell meter. RESULTS: Topical instillation of isopropyl unoprostone induced aqueous flare elevation in rabbit eyes. Also, topical isopropyl unoprostone additionally induced aqueous flare elevation in eyes with transcorneal diffusion of PGE(2). Latanoprost did not induce flare elevation. CONCLUSION: Isopropyl unoprostone induced aqueous flare elevation in rabbits, and latanoprost did not produce aqueous flare elevation. PMID- 11914612 TI - Ocular volume change per cardiac cycle. AB - Ocular pneumoplethysmograph (OPG) tests were performed in 499 employees of an industrial firm. The ocular volume change (OVC), in microliters per cardiac cycle, was calculated from each of OPG tests. In the 195 females, the OVC was 294 +/- 81 microl. In the 304 males, the OVC was 269 +/- 86 microl. The female-male OVC difference was significant, p = 0.001. PMID- 11914614 TI - Age-related macular degeneration: comments on the articles by E.L. Altschuler and R.A. Weale. PMID- 11914613 TI - In vitro toxicity of rivastigmine and donepezil in cells of epithelial origin. AB - Neurospecific acetylcholinesterase inhibitors have been shown to lower intraocular pressure (IOP) in normal rabbits and might have additional neuroprotective effects. This study was set out to explore and compare the toxicity of two selective acetylcholinesterase inhibitors, rivastigmine and donepezil, on two standardized cell lines of epithelial origin. Chang and Vero cells were incubated with various concentrations of rivastigmine or donepezil. Acute toxicity (4 h) was assessed by monitoring the permeability of cells to propidium iodide. Chronic toxicity (7 days) was determined by monitoring the effect of the two drugs on esterase activity and cell proliferation. The viability of cells was also assessed morphologically by microscopic inspection. Signs of acute toxicity became manifest at a rivastigmine concentration of 50 mg/ml in both Chang and Vero cells. Indications of chronic toxicity became obvious at concentrations of as low as 1 x 10(-5) mg/ml. In contrast, degenerative morphological changes became manifest only at a concentration of as high as 1 mg/ml. In donepezil-treated cells, acute toxicity was not observed in the concentration range tested, whereas chronic toxicity was detected at 1 x 10( 1) mg/ml in both Chang and Vero cells, a concentration at which degenerative morphological changes became evident as well. In contrast to rivastigmine, donepezil elicited no signs of acute or chronic toxicity in either Chang or Vero cells at the IOP-lowering concentration of 1 x 10(-4) mg/ml. At this dose, the drug is therefore unlikely to evoke deleterious effects on ocular surface tissues in the clinical setting. PMID- 11914615 TI - Antisilicone antibodies in patients with silicone implants for retinal detachment surgery. PMID- 11914618 TI - Solid variant of mammary adenoid cystic carcinoma with basaloid features: a study of nine cases. AB - Adenoid cystic carcinoma of the breast is a rare neoplasm that represents <1% of breast carcinomas. The tumors are histologically indistinguishable from examples in other sites, and they have a generally favorable prognosis. Several studies have investigated the possible correlation between histologic grade in adenoid cystic carcinoma (largely determined by cytology and growth pattern) and prognosis. Some earlier reports concluded that a solid variant of mammary adenoid cystic carcinoma had a more aggressive clinical course, but others did not confirm this impression. This report describes nine patients with a solid variant of mammary adenoid cystic carcinoma that has a striking basaloid appearance. All were women ranging in age from 37 to 83 years. A solitary mass was evident in all patients. Tumor size was 1.1-15 cm (mean 3.7 cm). The tumors exhibited a predominantly solid architecture comprised of basaloid appearing cells with moderate to marked nuclear atypia. Five tumors had >5 mitotic figures per 10 high power microscopic fields. Intercalated ducts were found in all tumors, being well formed in six and poorly formed in three. Immunohistochemical stains for cytokeratins, basement membranes, and vimentin were consistently positive. Surgery was performed in all cases consisting of excision in seven and mastectomy in two. Axillary lymph node metastases were found in two of six axillary dissections and four had negative lymph nodes. The lymph nodes were not examined in three patients. Follow-up information was available for seven patients. Six women had no evidence of disease after follow-up of 2-88 months (mean 32 months), one patient died of unknown causes, and one patient was lost to follow-up. It is concluded that the solid variant of mammary carcinoma with basaloid features is a histologically distinct tumor that is capable of axillary metastases. Long-term follow-up of a larger series of cases will be needed to determine whether the prognosis of these patients differs significantly from that of women with conventional adenoid cystic carcinoma. Presently, these patients are candidates for axillary staging by sentinel lymph node mapping or low axillary dissection if there is no clinical evidence of axillary metastases. Systemic adjuvant treatment would be prudent when axillary nodal metastases are present. Breast-conserving surgery with radiation is an option if negative margins can be achieved because this appears to be a unicentric form of carcinoma. PMID- 11914617 TI - An immunohistochemical analysis of endometrial stromal and smooth muscle tumors of the uterus: a study of 54 cases emphasizing the importance of using a panel because of overlap in immunoreactivity for individual antibodies. AB - The expression of desmin, h-caldesmon, calponin, CD10, CD34, CD99, inhibin, and keratin (AE1/3-Cam 5.2) was studied in 10 conventional leiomyomas, 9 highly cellular leiomyomas, 9 epithelioid smooth muscle tumors, 9 leiomyosarcomas, 10 endometrial stromal tumors (4 with smooth muscle metaplasia), and 7 uterine tumors resembling ovarian sex cord tumors (UTROSCTs). c-kit expression was tested in 10 endometrial stromal tumors, 7 UTROSCTs, and 9 leiomyosarcomas. Desmin was positive in almost all smooth muscle tumors except those of epithelioid type, which were positive in only about half of the cases. It also stained areas of smooth muscle differentiation in endometrial stromal tumors and five of seven UTROSCTs. h-caldesmon was positive in almost all nonepithelioid smooth muscle tumors and in areas of smooth muscle differentiation in endometrial stromal tumors; it was positive in only about half of the epithelioid smooth muscle tumors and negative in all UTROSCTs. Calponin was positive in most tumor types. CD10 was positive in nine of 10 endometrial stromal tumors and five of seven UTROSCTs, although very focally in the latter group. It was also expressed, however, in almost all leiomyosarcomas, almost 50% of highly cellular leiomyomas, and rarely in the other smooth muscle tumors. CD34 was negative in the tested tumors with rare exceptions. CD99 and inhibin were positive in four of seven and one of seven UTROSCTs. Keratin positivity was found in most (five of seven) UTROSCTs and occasionally in smooth muscle tumors (seven of 37). c-kit was negative in all endometrial stromal tumors, UTROSCTs, and leiomyosarcomas. The major conclusions of this study are as follows: 1) Pure endometrial stromal tumors are usually desmin negative. 2) In contrast to some previous studies, CD10 expression was often seen in smooth muscle tumors, including most leiomyosarcomas and almost half of highly cellular leiomyomas. As a result, a panel of CD10, h caldesmon, and desmin should be used and will distinguish endometrial stromal tumors from highly cellular leiomyomas in most cases. 3) In contrast to a previous study, no significant differences in immunoreactivity were seen between h-caldesmon and desmin in tumors with smooth muscle differentiation. 4) The absence of h-caldesmon in UTROSCTs helps separate them from epithelioid smooth muscle tumors. 5) UTROSCTs may express epithelial, stromal, and smooth muscle markers, suggesting divergent differentiation. 6) Our study shows less frequent inhibin expression in the sex cord-like elements of the UTROSCTs than in other studies. 7) c-kit may help distinguish metastatic endometrial stromal tumors of the uterus (c-kit negative) from gastrointestinal stromal tumors (c-kit positive). 8) CD34, CD99, and keratin have no or minimal role in this area, but keratin positivity in smooth muscle tumors should not lead to their confusion with epithelial tumors. PMID- 11914619 TI - Histopathologic types of benign breast lesions and the risk of breast cancer: case-control study. AB - The hypothetical multistep model of carcinogenesis indicates that breast cancer develops via a series of intermediate hyperplastic lesions through in situ to invasive carcinoma. To identify the risk inherent within the different morphologic lesions, we have analyzed the data from 674 benign biopsy specimens comprising 120 cases who subsequently developed breast cancer and 382 controls (matched for age and date of biopsy) spanning a period up to 20 years of follow up (mean 66.95 months). In this series we have confirmed an increased risk associated with certain types of benign breast lesions. Atypical lobular hyperplasia was the most significant risk factor for breast cancer with more unfavorable outcome in patients <50 years of age (p = 0.003) and a relative risk (RR) of 4.55 (confidence interval [CI] 1.77-11.69). Hyperplasia of usual type showed an RR of 1.53 (CL 1.10-2.13) with a statistically worse probability of survival (cancer-free time) for patients >50 years. For atypical ductal hyperplasia the RR was 2.03 (CI 0.80-1.39). Blunt duct adenosis was significantly more common in cases progressing to breast cancer compared with controls, showing an RR of 2.08 (CI 1.12-2.85). We describe in detail the criteria of morphologic changes observed in blunt duct adenosis and define, for the first time, the level of risk associated with each of its six subtypes. Improved knowledge of breast carcinogenesis will provide insight for defining high-risk groups thus resulting in improved screening and management regimens. PMID- 11914620 TI - Perineural invasion and MIB-1 positivity in addition to Gleason score are significant preoperative predictors of progression after radical retropubic prostatectomy for prostate cancer. AB - We assessed the use of clinical stage, serum prostate specific antigen, DNA ploidy, proliferation, and traditional histologic findings from the biopsy to predict prostate cancer progression after radical retropubic prostatectomy. Between 1995 and 1998, 454 consecutive patients with cancer on biopsy were treated by radical retropubic prostatectomy. Preoperative serum prostate specific antigen, clinical stage, Gleason score, percentage of cores and surface area positive for cancer, perineural invasion, and DNA ploidy and MIB-1 immunostain quantitation by image analysis were evaluated in a multivariate Cox proportional hazards regression model to predict cancer progression. Cancer progression was defined as a postoperative serum prostate specific antigen level of > or = 0.4 ng/mL, local recurrence, or systemic progression. Mean follow-up was 3.4 years (range 17 days to 5.8 years). Cancer progression was observed in 73 patients with a mean time to progression of 2.1 years (range 33 days to 5.1 years). Gleason score (p <0.001), MIB-1 cancer proliferation (p = 0.008), and perineural invasion (p = 0.008) were significantly associated with progression. Patients with cancer Gleason scores of 7 and >7 had a 2.5-fold and nearly 4-fold increased risk, respectively, of cancer progression compared with patients with cancer Gleason scores of < or = 6. Patients with perineural invasion at biopsy were twice as likely to progress compared with patients without perineural invasion. Each 1 unit increase in MIB-1 on the natural logarithmic scale increased the risk of cancer progression by 64%. Cancer progression models that include serum prostate specific antigen and clinical stage may require revision to incorporate perineural invasion and MIB-1 proliferative activity in addition to Gleason score. PMID- 11914621 TI - Endometrial stromal sarcoma metastatic to the lung: a detailed analysis of 16 patients. AB - Pulmonary metastases of endometrial stromal sarcoma (ESS) are uncommon and can pose diagnostic problems. We reviewed lung specimens from 16 patients with metastatic ESS. Patients were 31-77 years of age at the time of lung biopsy. Uterine ESSs were diagnosed an average of 9.8 years before lung biopsy in 11 patients. Uterine ESSs were originally called smooth muscle tumors in three additional patients. Thirteen patients were evaluated for new pulmonary nodules, seven of whom were asymptomatic. Nodules were multiple in 14 and solitary in four, ranging from 1.0 to 8.0 cm in greatest dimension. One patient died of metastatic disease; 14 were alive and seven of these were without disease (mean follow-up 4.1 years). Diagnostic considerations in 12 consultation cases included ESS, sclerosing hemangioma, carcinoid tumor, lymphangioleiomyomatosis, endometriosis, hemangiopericytoma, and lymphoma. Tumors were well circumscribed and usually solid, composed of plump spindle cells arranged in short fascicles. Two tumors were predominantly cystic. Sex cord-like stromal differentiation was identified in three. Neoplastic cells stained for vimentin (93%), estrogen and progesterone receptor (100%), smooth muscle actin (57%), desmin (50%), and keratin (46%). Metastatic ESS should be included in the differential diagnosis of nonepithelial neoplasms in women. PMID- 11914622 TI - Hypopigmented mycosis fungoides: frequent expression of a CD8+ T-cell phenotype. AB - Hypopigmented mycosis fungoides (MF) is a form of cutaneous T-cell lymphoma in which hypopigmentation occurs in the absence of classic lesions of MF. Hypopigmented MF predominantly affects people with dark complexions. The natural history of this variant of cutaneous T-cell lymphoma is similar to that of conventional MF, although the disease onset is usually in childhood or adolescence. In a retrospective study we evaluated the clinical, histopathologic, immunohistochemical, and molecular characteristics of hypopigmented MF in 15 patients. Similar to other reports, the disease onset occurred in childhood and adolescence in most of the cases. The survival rate was comparable with that of classic MF. We did not observe progression to systemic disease or lymph node involvement. Histopathologically hypopigmented lesions were indistinguishable from hyperpigmented or erythematous patches. On immunohistochemical analysis a predominantly CD8+ infiltrate was detected in the majority of cases (nine of 15 patients). To determine whether epidermotropic CD8+ T cells represent the malignant T-cell clone or whether these cells are innocent, tumor-infiltrating T lymphocytes, we performed microdissection of epidermotropic CD8+ T cells and analyzed T-cell receptor-gamma chain gene for rearrangements. The epidermotropic CD8+ T lymphocytes showed clonal T-cell receptor gene rearrangement and therefore represented the malignant T-cell clone. We conclude that hypopigmented MF tends to occur in young people and that it belongs to the group of CD8+ cutaneous T cell lymphomas in the majority of cases. PMID- 11914623 TI - Oncocytic adenocarcinoma of the stomach: parietal cell carcinoma. AB - We report 10 cases of an unusual type of gastric adenocarcinoma that occurred in elderly patients 58-81 years of age. Histologically, the tumors were well to moderately differentiated tubular adenocarcinomas with very eosinophilic, finely granular cytoplasm. Immunohistochemical stains for antimitochondrial antibody were strongly positive. Ultrastructurally, the tumor cells had numerous mitochondria in their cytoplasm and occasional intracytoplasmic lumina with associated long microvilli. These histologic and ultrastructural features are similar to those of parietal cells in normal gastric fundic mucosa, but immunohistochemical staining of the tumors using four different antiparietal cell antibodies (anti-H(+)-K(+)-adenosine triphosphatase antibodies) was negative in all cases. Therefore, we think that these tumors were not parietal cell carcinomas but could be termed oncocytic adenocarcinomas, or adenocarcinomas with oncocytic differentiation. Previously reported cases of parietal cell carcinoma have been said to have a favorable prognosis, but it will be necessary to study a larger number of cases to determine the prognosis of oncocytic adenocarcinoma. PMID- 11914624 TI - The mucin profile of noninvasive and invasive mucinous cystic neoplasms of the pancreas. AB - Recently, it was shown that ductal adenocarcinomas and intraductal papillary mucinous neoplasms of the pancreas differ in their expression of the mucin markers MUC1 and MUC2 while both tumors express MUC5AC. It is not known whether mucinous cystic neoplasms of the pancreas have their own mucin profile. To clarify this issue, 22 mucinous cystic neoplasms were examined immunohistologically for their expression of MUC1, MUC2, MUC5AC, and MUC6 and also for the protein products of the tumor suppressor genes p53 and DPC4 and the mismatch repair genes. Noninvasive mucinous cystic neoplasms, regardless of the degree of cellular atypia, were all positive for MUC5AC and negative for MUC1, with the exception of the cyst-lining epithelium of a single case with eosinophilic cytology (case no. 16). Only in cases with an invasive component was MUC1 expression observed. MUC2 expression was restricted to goblet cells scattered within the epithelium of the mucinous cystic neoplasms and was often accompanied by endocrine cells, a further indication of intestinal differentiation. DPC4 expression was maintained in all tumors, except for three invasive carcinomas. p53 nuclear reactivity was found in one borderline tumor and four invasive mucinous cystic carcinomas. The results suggest that the epithelium of noninvasive mucinous cystic neoplasms does not differ in its expression of MUC5AC from ductal adenocarcinomas, intraductal papillary-mucinous neoplasms, and metaplastic pancreatic duct epithelium. The fact that noninvasive mucinous cystic neoplasms lack MUC1 expression (except for an eosinophilic variant) but express it when they become invasive might be used as a marker indicating the step of progression from noninvasiveness to invasiveness. PMID- 11914625 TI - Evaluation of molecular markers in low-grade diffuse astrocytomas: loss of p16 and retinoblastoma protein expression is associated with short survival. AB - Diffuse astrocytomas have a median survival of 6-8 years. However, in a minority of cases that are histologically low grade, progression is rapid, leading to death within 2 years. Loss of p16, retinoblastoma protein, and deleted-in-colon carcinoma protein expression, and monosomy of chromosome 10 have been shown to occur in malignant astrocytic tumors. We have investigated the prognostic value of expression of these markers, using techniques applicable in many histopathology laboratories, in diffuse astrocytomas that are histologically low grade. Paraffin sections from 71 diffuse, supratentorial, low-grade astrocytomas, from patients with at least 8-year survival data, were immunostained with antibodies to p16, deleted-in-colon-carcinoma protein, p53, Ki67, and retinoblastoma protein. In situ hybridization with a digoxigenin-labeled probe to chromosome 10 was used to assess chromosomal loss. In most cases there was immunostaining of virtually all tumor cell nuclei with antibodies to p16 and retinoblastoma protein. Three of the 68 tumors in which assessment of p16 was possible included discrete foci with lack of detectable immunoreactivity in tumor cells. The three patients concerned had a significantly shortened median survival (1.1 years vs 4.4 years in those without loss of p16; p <0.01). In six of the 61 cases where assessment of retinoblastoma protein was possible, <70% of tumor cell nuclei showed immunoreactivity. These six patients had a shorter survival (4.0 years) than had the remaining patients (5.4 years), although this difference was not statistically significant. The tumor from one of these patients included areas where only 36% of tumor cells showed retinoblastoma protein immunoreactivity, and this patient survived only 1.5 years. Tumors showing loss of both p16 and retinoblastoma were not seen. p53 and deleted-in-colon-carcinoma protein expression was highly variable and did not correlate with survival. Tumors with monosomy for chromosome 10 were not identified. Both polyploidy and the Ki67 labeling index were significantly associated with the p53 labeling index but not with survival. Focal loss of p16 or retinoblastoma protein is demonstrable in approximately 5% and 10% of diffuse low-grade diffuse astrocytomas, respectively. Tumors with focal loss of immunoreactivity for these proteins are associated with shorter survival than those without, suggesting that immunohistochemistry for p16 and retinoblastoma protein may be a useful adjunct to other methods for assessing the prognosis of astrocytomas. PMID- 11914626 TI - Immunophenotype of pleomorphic xanthoastrocytoma. AB - Pleomorphic xanthoastrocytoma (PXA) is an uncommon tumor, often seizure associated and occurring in the temporal lobe of young adults. Although its cells are considered astrocytic in nature, recent studies suggest the presence of neuronal differentiation and a possible relationship to glioneuronal neoplasms. We immunostained 40 cases of PXA, including two composite PXA-gangliogliomas (PXA GG), with a panel of glial (glial fibrillary acidic protein, S-100 protein) and neuronal markers (class III beta-tubulin, synaptophysin, neurofilament proteins, MAP2, and chromogranin A). Conventional PXAs demonstrated immunoreactivity for glial fibrillary acidic protein (100% of cases), S-100 protein (100%), class III beta-tubulin (73%), synaptophysin (38%), NF proteins (18 and 8%), and MAP2 (8%). Chromogranin A stain was absent in all conventional PXA cases. Neoplastic ganglion cells in both PXA-GGs stained with class III beta-tubulin, synaptophysin, and chromogranin A. Ultrastructural studies, performed in nine cases, demonstrated neuronal features including microtubules, dense core granules, and/or clear vesicles largely limited to cell processes (two PXAs) and in the cytoplasm (PXA component of one PXA-GG). Although the essential nature of PXA is clearly and uniformly glial, the significance of the limited neuronal differentiation is unclear, as it is the relationship between conventional PXA and PXA-GG. We found no evidence that the former is a precursor of the latter. PMID- 11914627 TI - C-kit expression in pediatric solid tumors: a comparative immunohistochemical study. AB - The stem cell factor/c-kit tyrosine kinase receptor pathway has been shown to be important for tumor growth and progression in several cancers, including mast cell diseases, gastrointestinal stromal tumor, acute myeloid leukemia, small cell lung carcinoma, and Ewing sarcoma. Studies using the oral agent STI-571 (Gleevec, Novartis), an inhibitor of the tyrosine kinases bcr-abl, c-kit, and PDGFR, have shown significant responses in patients with chronic myelogenous leukemia and gastrointestinal stromal tumor. With the aim of identifying additional groups of tumors that may use the stem cell factor/c-kit pathway and secondarily may be responsive to STI-571 treatment, this study surveyed 151 primary tumors from patients treated at St. Jude Children's Research Hospital for immunohistochemical expression of c-kit. Formalin-fixed, paraffin-embedded sections were stained with rabbit polyclonal anti-human c-kit (CD117, Dako) using standard avidin-biotin peroxidase complex technique, antigen retrieval, and an automated stainer. Strong, diffuse staining for c-kit was seen in a proportion of synovial sarcomas, osteosarcomas, and Ewing sarcomas. Strong, diffuse staining was less common in neuroblastomas, Wilms' tumors, and rhabdomyosarcomas and was negative in alveolar soft part sarcomas and desmoplastic small round cell tumors. Tumors with strong, diffuse staining for c-kit in a pattern similar to gastrointestinal stromal tumor may represent suitable targets for new therapeutic agents. PMID- 11914628 TI - Expression of KIT (CD117) in angiomyolipoma. AB - Angiomyolipoma is a unique mesenchymal tumor postulated to arise from perivascular epithelioid cells. Immunohistochemical studies have shown that angiomyolipomas express the melanocytic markers HMB-45, MART-1 (Melan A), microphthalmia transcription factor, and tyrosinase, in addition to smooth muscle actin. KIT (CD117) is a transmembrane growth factor receptor expressed in cells of melanocytic and a variety of other cell lineages. To date, KIT immunoreactivity has not been systematically studied in angiomyolipoma. In this study we immunohistochemically analyzed a series of 21 angiomyolipomas (15 hepatic, six renal) with KIT. All were KIT positive: 14 of 21 (67%) with 3+ staining (>50% of tumor cells), 4 of 21 (19%) with 2+ staining (25-50% of tumor cells), and 3 of 21 (14%) with 1+ staining (<25% of tumor cells). In comparison, the percent of angiomyolipomas showing 3+ staining with HMB-45 was 62% and with Melan A was 52%. Positive KIT staining was detected in the epithelioid, spindle, and intermediate small round cells. Most cases showed diffuse cytoplasmic positivity. Strong perinuclear staining was present in the vacuolated clear epithelioid cells. There was focal KIT staining of fat cells. KIT was not detected in the endothelial cells lining blood vessels within the tumor. KIT may be a useful ancillary marker for the diagnosis of angiomyolipoma. Angiomyolipoma should be included in the differential diagnosis of KIT-positive tumors. PMID- 11914629 TI - Acute radiation colitis in patients treated with short-term preoperative radiotherapy for rectal cancer. AB - The histopathologic features of acute radiation-induced colitis in humans have been described in occasional, >20-year-old studies, but they have not been analyzed in detail. We characterize such findings in 34 patients with rectal cancer who underwent surgery a few days after preoperative irradiation with 25 Gy given over 5-7 days, and we compare the results to the histopathologic features detected in 18 patients treated by a conventional preoperative irradiation protocol consisting of 45 Gy during 5 weeks followed by surgery after a time interval of at least 3 weeks. Short-term preoperative irradiation therapy generally induced severe mucosal inflammation characterized by increased cellularity of the lamina propria, prominent eosinophilic infiltrates, crypt disarray, surface and crypt epithelial damage, nuclear abnormalities, and presence of apoptotic bodies in the crypt epithelium. These histopathologic features were absent or detected only occasionally in the patient group treated according to the long-term preoperative irradiation protocol. Despite acute severe inflammation, none of the patients treated by short-term irradiation developed perioperative complications. These observations indicate that acute radiation colitis may remain clinically silent and resolve spontaneously within a few weeks after irradiation. Given the widening acceptance of short-term preoperative irradiation protocols for rectal cancer, pathologists should be aware of the rather characteristic histologic findings of acute radiation colitis and avoid unnecessary concern of clinicians. The differential diagnosis includes infectious colitis, collagenous and ischemic colitis, nonsteroidal anti inflammatory drug-associated colitis, and chronic idiopathic inflammatory bowel disease. PMID- 11914630 TI - Endometrial polyps with atypical (bizarre) stromal cells. AB - Atypical stromal cells of the lower female gynecologic tract have been specifically described in the vagina, vulva, and cervix, predominantly in the context of polyps. Rare cases of atypical stromal cells have been documented in the endometrium. We report a series of 15 examples of atypical stromal cells in the endometrium: 13 in endometrial polyps and two within endometrial stroma in curettage/biopsy specimens unassociated with polyps. The patients ranged in age from 45 to 82 years. Immunohistochemical studies were performed to aid in the identification of the origin of these atypical cells. The differential diagnoses included adenosarcoma, endometrial stromal sarcoma, and, less likely, malignant mesodermal mixed tumor (MMMT/carcinosarcoma). Similar to atypical stromal cells reported in other gynecologic sites, such cells discovered in the endometrium also appear to have a benign clinical course after complete excision or polypectomy (follow-up ranging from 1 month to 44 months). Accurate recognition of this lesion is essential to avoid unnecessary surgical overtreatment. Because of their rarity, limited available data, and lack of significant long-term follow up, continued clinical monitoring of these patients would be prudent. PMID- 11914631 TI - Molecular pathogenesis of pulmonary carcinosarcoma as determined by microdissection-based allelotyping. AB - Pulmonary carcinosarcoma is a rare, biphasic tumor composed of malignant epithelial and mesenchymal elements. Its histogenesis is controversial in light of the presence of divergent cell lineages and the clonal nature of malignancy. To address these issues, we performed an extensive comparative genotypic analysis using microdissection to secure representative mesenchymal and epithelial components from each of six cases of pulmonary carcinosarcoma. Loss of heterozygosity was analyzed with a panel of 12 polymorphic microsatellite markers designed to indicate allelic loss and situated in proximity to known tumor suppressor genes located on 1p, 3p, 5q, 9p, 10q, and 17p. In accordance with the relatively greater biologic aggressiveness of this tumor type, both the epithelial and mesenchymal components showed extensive allelic loss, most notably for 3p, 5q, and 17p. More importantly, we found overall equivalent patterns of acquired allelic loss between the two components on an individual case basis, strongly supporting the monoclonal origin of these neoplasms. Minor differences in the allelic fingerprint between the two cell lineages could be explained by progressive accumulation of allelic loss alterations that appear to occur more frequently in the mesenchymal component of the tumor. The data support the efficacy of microdissection-based allelic fingerprinting to delineate the relationship between different morphologic components of a single neoplasm. PMID- 11914632 TI - Carcinoid tumor of the esophagus: a clinicopathologic study of four cases. AB - Several case reports have emphasized that esophageal carcinoid tumors are associated with a poor prognosis. To expand our knowledge about the pathology and biologic behavior of these rare tumors, we reviewed the clinicopathologic and immunohistochemical findings of four cases of primary esophageal carcinoid. The age of the patients ranged from 48 to 82 years (mean 63 years; median 61 years). The lower segment of the esophagus was involved in two cases and the mid segment was involved in one case. The sizes of the tumors ranged from 0.3 cm to 3.5 cm. Two tumors were confined to the lamina propria and two invaded into the muscular wall. Two tumors appeared polypoid, whereas the remaining two were incidental findings and associated with adenocarcinoma arising in a background of Barrett esophagus. The adenocarcinoma was superficially invasive in one case, whereas it penetrated the muscular wall in the other. All four carcinoid tumors were immunoreactive with chromogranin and synaptophysin. There was focal expression of serotonin in two cases, glucagon in one case, and pancreatic polypeptide in one case. Endocrine cell hyperplasia was noted in both the Barrett esophagus and the invasive adenocarcinoma. One patient died secondary to postoperative pneumonia. Three patients are alive and disease free at 1, 6, and 23 years status post therapy. None of the patients had metastatic disease. These findings show that esophageal carcinoids are associated with a favorable prognosis. They arise in two settings: (1) a single large polypoid tumor or (2) an incidental finding and in association with adenocarcinoma arising in the background of Barrett esophagus. The presence of endocrine cell hyperplasia in the Barrett mucosa and the adenocarcinoma supports the hypothesis that these lesions arise from a common stem cell. PMID- 11914633 TI - Malignant transformation of renal angiomyolipoma: a case report. AB - We report a case of renal angiomyolipoma (AML) with malignant transformation. A 28-year-old woman developed large bilateral renal masses 5 months before admission to our hospital. She was diagnosed with tuberous sclerosis when she was 4 years old. Total nephrectomy of the left kidney was performed, but she died during the operation. Although the focal region of the resected tumor had the appearance of a classic AML, most of the lesion showed a diffuse proliferation of atypical epithelioid cells resembling that in renal cell carcinoma. The epithelioid cells had extremely pleomorphic and hyperchromatic nuclei with frequent mitotic figures, including atypical forms. Immunohistochemical analysis revealed that the atypical epithelioid cells and the typical AML lesions were both positive for HMB-45 but that the former were negative for epithelial and myogenic markers. The smooth muscle cells and thick-walled vessels were focally positive for muscle-specific actins. Furthermore, the atypical epithelioid cells were immunoreactive for p53, whereas the foci of the typical AML were negative. Examination of the microdissected paraffin-embedded tissues revealed p53 mutations in the malignant epithelioid areas in AML but not in the renal parenchyma or typical AML areas. In this case it is proposed that p53 mutation may play an important role in malignant transformation of renal AML. PMID- 11914634 TI - Interdigitating dendritic cell sarcoma of the spleen: report of a case with a review of the literature. AB - Interdigitating dendritic cell sarcoma is an extremely rare neoplasm that mainly occurs in the lymph nodes. We report a case of interdigitating dendritic cell sarcoma arising from the spleen, a previously unreported site for interdigitating dendritic cell sarcoma. An 87-year-old woman, visiting Ashigara Hospital with complaints of palpitation and dyspnea, was found to have pancytopenia and low proteinemia. Abdominal ultrasonography and CT scanning demonstrated severe splenomegaly with heterogeneous enhancement. She received a splenectomy under the clinical diagnosis of a splenic tumor. Grossly, the spleen was markedly enlarged, with confluent massive nodules. Microscopically, the normal architecture was effaced with diffuse proliferation of large pleomorphic cells arrayed in a somewhat sheet-like pattern. Erythrophagocytosis was commonly observed. Immunohistochemical studies showed that the tumor cells were positive for S-100 protein, fascin, vimentin, and CD68, but uniformly negative for CD45, B- and T cell markers, CD1a, CD30, complement receptors, CD34, Factor VIII, HMB-45, and lysozyme. Ultrastructurally, the tumor cells possessed complex interdigitating cytoplasmic dendritic processes. Birbeck granules were absent. Based on these findings, the present case was diagnosed as interdigitating dendritic cell sarcoma. The patient died of multiple liver metastases 3 months postoperatively. PMID- 11914635 TI - Simple and complex hyperplastic papillary proliferations of the endometrium. PMID- 11914636 TI - Histone deacetylase inhibitors in cancer treatment. AB - Histone deacetylase (HDAC) inhibitors are emerging as an exciting new class of potential anticancer agents for the treatment of solid and hematological malignancies. In recent years, an increasing number of structurally diverse HDAC inhibitors have been identified that inhibit proliferation and induce differentiation and/or apoptosis of tumor cells in culture and in animal models. HDAC inhibition causes acetylated nuclear histones to accumulate in both tumor and normal tissues, providing a surrogate marker for the biological activity of HDAC inhibitors in vivo. The effects of HDAC inhibitors on gene expression are highly selective, leading to transcriptional activation of certain genes such as the cyclin-dependent kinase inhibitor p21WAF1/CIP1 but repression of others. HDAC inhibition not only results in acetylation of histones but also transcription factors such as p53, GATA-1 and estrogen receptor-alpha. The functional significance of acetylation of non-histone proteins and the precise mechanisms whereby HDAC inhibitors induce tumor cell growth arrest, differentiation and/or apoptosis are currently the focus of intensive research. Several HDAC inhibitors have shown impressive antitumor activity in vivo with remarkably little toxicity in preclinical studies and are currently in phase I clinical trial. The focus of this review is the development and clinical application of HDAC inhibitors for the treatment of cancer. PMID- 11914637 TI - In vitro and in vivo characterization of XR11576, a novel, orally active, dual inhibitor of topoisomerase I and II. AB - XR11576, a novel phenazine, was developed as an inhibitor of both topoisomerase I and II. This study characterized the ability of XR11576 to inhibit both enzymes, and determined its in vitro and in vivo antitumor efficacy against a number of murine and human tumor models. XR11576 was a potent inhibitor of purified topoisomerase I and IIalpha, and exhibited similar potency for both enzymes. The compound stabilized enzyme-DNA cleavable complexes indicating that it acted as a topoisomerase poison. The DNA cleavage patterns obtained with XR11576 were different from those induced by camptothecin and etoposide, which are topoisomerase I and II poisons, respectively. XR11576 demonstrated potent cytotoxic activity against a variety of human and murine tumor cell lines (IC50=6 47 nM). Its activity profile was comparable to or better than that of many widely used anticancer drugs. Moreover, XR11576 was unaffected by multidrug resistance (MDR) mediated by overexpression of either P-glycoprotein or MDR-associated protein, or by down-regulation of topoisomerase II. The latter property supports the dual inhibitory mechanism of action of the compound. XR11576 exhibited a similar pharmacokinetic profile in mice and rats after either i.v. or p.o. administration. In vivo XR11576 showed marked efficacy against a number of tumors including sensitive (H69/P) and multidrug-resistant (H69/LX4) small cell lung cancer and the relatively refractory MC26 and HT29 colon carcinomas following i.v. and p.o. administration. The efficacy of XR11576 was at least comparable to that of TAS-103, originally proposed as a dual inhibitor of topoisomerase I and II. These results suggest that XR11576 is a promising new antitumor agent with oral and i.v. activity, and warrants further development. PMID- 11914638 TI - Nucleoside transport inhibitors, dipyridamole and p-nitrobenzylthioinosine, selectively potentiate the antitumor activity of NB1011. AB - NB1011, a novel anticancer agent, targets tumor cells expressing high levels of thymidylate synthase (TS). NB1011 is converted intracellularly to bromovinyldeoxyuridine monophosphate (BVdUMP) which competes with the natural substrate, deoxyuridine monophosphate, for binding to TS. Unlike inhibitors, NB1011 becomes a reversible substrate for TS catalysis. Thus, TS retains activity and converts BVdUMP into cytotoxic product(s). In vitro cytotoxicity studies demonstrate NB1011's preferential activity against tumor cells expressing elevated TS protein levels. Additionally, NB1011 has antitumor activity in vivo. To identify drugs which interact synergistically with NB1011, we screened 13 combinations of chemotherapeutic agents with NB1011 in human tumor and normal cells. Dipyridamole and p-nitrobenzylthioinosine (NBMPR), potent inhibitors of equilibrative nucleoside transport, synergized with NB1011 selectively against 5 fluorouracil (5-FU)-resistant H630R10 colon carcinoma cells [combination index (CI)=0.75 and 0.35] and Tomudex-resistant MCF7TDX breast carcinoma cells (CI=0.51 and 0.57), both TS overexpressing cell lines. These agents produced no synergy with NB1011 in Det551 and CCD18co normal cells (CI > 1.1) lacking TS overexpression. Dipyridamole potentiated NB1011's cytotoxicity in medium lacking nucleosides and bases, suggesting a non-salvage-dependent mechanism. We demonstrate that nucleoside transport inhibitors, dipyridamole and NBMPR, show promise for clinically efficacious combination with NB1011. PMID- 11914639 TI - Growth inhibition of human melanoma tumor cells by the combination of sodium phenylacetate (NaPA) and substituted dextrans and one NaPA-dextran conjugate. AB - We have studied the cytostatic effects of sodium phenylacetate (NaPA) in association with several substituted dextrans on human tumor melanoma 1205LU cells. We show that NaPA alone inhibits the growth of these cells (IC50 = 3.9 mM) while a weak inhibitory effect appears at a concentration of 37 microM (10 microg/ml) for a dextran methyl carboxylate benzylamide (LS17-DMCB). The precursors of LS17-DMCB [T40 Dextran and carboxymethyl dextran (LS17-DMC)] did not affect the growth of 1205LU cells. To potentiate the inhibitory activity of NaPA at low concentrations (below 5.6 mM), we have tested NaPA and LS17-DMCB in physical mixture (association) or linked together covalently (this conjugate is termed 'LS17-NaPaC'). We have observed an increase of the 1205LU cell growth inhibition effect with NaPA in association (IC50 1.8 mM). For a concentration of 5 mM of NaPA (free in the case of association or linked in the case of conjugate), the association with dextran derivative exhibits a 4.6-fold higher efficacy than with NaPA alone (9 versus 41% surviving fraction), while the conjugate is 1.3-fold smaller (52% growth inhibition). By performing isobologram analysis of the IC50 data, we have shown a synergistic effect for a particular molar ratio of NaPA and LS17-DMCB (NaPA:LS17-DMCB = 0.35). PMID- 11914640 TI - Efficient induction of apoptosis by ONYX-015 adenovirus in human colon cancer cell lines regardless of p53 status. AB - The ONYX-015 virus is a mutated adenovirus that in theory selectively replicates and induces cytolysis in tumor cells lacking functional p53. The present study investigated whether ONYX-015 viral infection alone or in combination with conventional chemotherapeutic agents could significantly increase apoptosis in human colon cancer cell lines, regardless of p53 status, compared to untreated cells. A pair of colon cancer cell lines that differ only in their p53 status (RKO with wild-type p53 and RKOp53 with deficient p53) was tested. Two chemotherapeutic agents, 5-fluorouracil (5-FU) and CPT-11, were tested in combination with ONYX-015. Final concentrations of these agents corresponded to peak plasma levels achievable in patients. ONYX-015 concentration was 10 p.f.u./cell. In RKO and RKOp53 cell lines, ONYX-015 viral infection alone or in combination with 5-FU or CPT-11 induced a significant increase in apoptosis compared to chemotherapeutic agents alone, regardless of p53 status. Moreover, the combination of ONYX-015 and chemotherapeutics induced more apoptosis than chemotherapeutics alone in the two colon cancer cell lines independently of their p53 status. We conclude that ONYX-015 virus infection alone or in combination with 5-FU or CPT-11 induced apoptosis in human colon cancer cell lines, independently of p53 status. PMID- 11914641 TI - Recombinant interleukin-2 treatment decreases P-glycoprotein activity and paclitaxel metabolism in mice. AB - Recombinant rIL-2 was reported to be able to decrease P-glycoprotein (P-gp) expression in cultured cells from human colon carcinoma. P-gp is considered an important factor in the control of Taxol efflux from tumor cells. Based on the premise that Taxol pharmacokinetic parameters could be modified as a result of diminished P-gp expression induced by recombinant interleukin (rIL)-2 and that this might elicit an interaction between the two drugs, we evaluated the pharmacokinetics of a novel strategy combining i.p. immunotherapy with rIL-2 and a cytotoxic agent, Taxol. Mice were allocated to two groups treated with rIL-2 (15 microg x 2/day from day 1 to 4) then Taxol (10 mg/kg i.p. day 5) or Taxol (10 mg/kg i.p.) alone (control group). The Taxol + rIL-2 combination provoked the development of ascites, presumably due to the presence of Cremophor EL in the Taxol preparation. Paclitaxel was measured in plasma and ascites by HPLC with UV detection. Paclitaxel pharmacokinetics were strongly modified by rIL-2 pretreatment. Compared to that observed in control mice, the apparent volume of distribution increased dramatically (Vd/F = 18.2 versus 4.1 l/kg) and the apparent plasma clearance decreased (Cl/F = 1.12 versus 1.66 l/h/kg). P-gp expression was determined in the liver, lung, intestine, brain and kidney in the two groups by immunodetection with the C219 anti-P-gp monoclonal antibody. A significant decrease in P-gp expression was observed in the intestine and in the brain in the rIL-2-pretreated mice as compared to controls. To study the functionality of P-gp, we compared digoxin (a model P-gp substrate) pharmacokinetics before and after pretreatment with rIL-2 (10 microg x 2/day from day 1 to 4), after a single 1 microg oral dose of digoxin used to quantify P-gp activity. Results showed a decrease in oral digoxin clearance after rIL-2 pretreatment indicating modified P-gp activity. We conclude that rIL-2 pretreatment is able to decrease P-gp activity and paclitaxel metabolism in vivo. This is the first study to demonstrate a decrease in P-gp activity and expression in organs such as the brain in vivo. A novel strategy combining immunotherapy with rIL-2 and a cytotoxic agent could potentially improve clinical results, particularly in brain cancer. PMID- 11914642 TI - Peroxisome proliferator-activated receptor gamma augments tumor necrosis factor family-induced apoptosis in hepatocellular carcinoma. AB - Proliferator-activated receptor gamma (PPARgamma) is a nuclear receptor, which mainly associates with adipogenesis, but also appears to facilitate cell differentiation or apoptosis in certain malignant cells. This apoptosis induction by PPARgamma is increased by co-stimulation with tumor necrosis factor (TNF) alpha-related apoptosis-inducing ligand (TRAIL), a member of the TNF family. In this study, we investigated the effect of PPARgamma on Fas-mediated apoptosis in hepatocellular carcinoma (HCC) cell lines. PPARgamma was expressed on all seven HCC cell lines and located in their nuclei. 15-Deoxy-Delta-12,14-prostaglandin J2 (15d- PGJ2), a PPARgamma ligand, inhibited cellular proliferation in HepG2, SK Hep1 or HLE cells, unlike pioglitazone, another PPARgamma ligand, which did not have a significant influence on proliferation of these cells. However, 15d-PGJ2 facilitated Fas-mediated HCC apoptosis that could not be induced by Fas alone. These results suggest that PPARgamma can augment TNF-family-induced apoptosis. PMID- 11914643 TI - Combined analysis of two phase II trials in patients with primary and advanced breast cancer with epidoxorubicin and docetaxel+granulocyte colony stimulating factor. AB - Anthracyclines and taxanes are to date the most active cytotoxic agents in the treatment of breast cancer, and a combination of these is therefore considered to result in the highest response rates in the neoadjuvant, as well as in palliative treatment. These two phase II studies aimed to evaluate the feasibility, toxicity and activity of a cytostatic regimen combining epidoxorubicin and docetaxel in outpatient patients suffering from breast cancer. In total, 104 consecutive patients were enrolled in these prospective clinical trials. The chemotherapeutic regimen consisted of epidoxorubicin [75 mg/m2 body surface area (BSA)] and docetaxel (75 mg/m2 BSA) on day 1 accompanied by the administration of granulocyte colony stimulating factor on days 3-10, repeated every 3 weeks (ED+G). Sixty-six patients received ED+G as neoadjuvant and 38 patients as palliative treatment, respectively. Patients received a total of 566 cycles (median: 6 cycles, range: 2-11 cycles) of this therapeutic regimen. Outpatient ED+G was well tolerated. A major response to preoperative ED+G could be demonstrated in 54 of 66 patients (82%) and in 22 of 38 palliative treated patients (58%). We conclude that outpatient ED+G is safe in the neoadjuvant and palliative treatment of patients suffering from breast cancer by showing a favorable side effect and activity profile. Thus, this regimen can be considered for further clinical trials. PMID- 11914644 TI - Implication of thymidylate synthase in the outcome of patients with invasive ductal carcinoma of the pancreas and efficacy of adjuvant chemotherapy using 5 fluorouracil or its derivatives. AB - Thymidine synthase (TS) is a key enzyme in the synthesis of pyrimidine in the de novo pathway of DNA synthesis and a major target of 5-fluorouracil (5-FU), but the implications of TS regarding human pancreatic cancer have not been reported. We assessed the expression of TS in invasive ductal carcinoma (IDC) of the pancreas by immunostaining and evaluated its clinicopathological significance, especially its implications regarding the efficacy of chemotherapy with 5-FU or its derivatives. The expression of TS in the nuclei of pancreatic cancer cells in 72 primary lesions of resectable IDC and 30 distant metastases of unresectable IDC was examined by immunostaining using anti-TS polyclonal antibody and immunoreactivity was classified into three categories: negative (-), low (+) and high (2+). High TS immunoreactivity was detected in 43% (31 of 72) of the primary lesions of the resectable IDCs and in 47% (18 of 38) of the metastatic lesions of the unresectable IDCs. The high TS in primary lesions showed a significantly inverse correlation with the level of nodal involvement. High TS immunoreactivity had a significant influence on the outcome of patients with resectable IDC and the rate of survival of the high TS immunoreactivity group was significantly higher than that of the negative or low reactivity groups, although high TS immunoreactivity did not have a significant influence on survival of the patients with unresectable IDC. The implications of TS immunoreactivity regarding the efficacy of 5-FU-based adjuvant chemotherapy (ACT) was also assessed. The high TS immunoreactivity group showed significantly better survival in both the patients who received ACT and those who were treated by surgery alone, in the resectable IDC among patients with resectable IDC. In cases of unresectable IDC, there were no differences in survival between the high and low TS groups among the patients who received ACT and those who were treated by surgery. In conclusion, high TS immunoreactivity was found to be cogent in predicting the prognosis of patients with pancreatic IDC, but its implications regarding the efficacy of 5-FU-based ACT are still unclear. PMID- 11914645 TI - Topoisomerase I levels in white blood cells of patients with ovarian cancer treated with paclitaxel-cisplatin-topotecan in a phase I study. AB - Topotecan stabilizes the topoisomerase I (Topo I) cleavable complex. We measured Topo I levels in white blood cells of patients with ovarian cancer treated with topotecan. Topotecan was given i.v. daily x 5 q 3 weeks in combination with paclitaxel (1 day before topotecan) and cisplatin (just prior topotecan). Our aim was to correlate Topo I levels to pharmacokinetics and toxicity. Topo I levels were determined using Western blotting and were expressed relative to the Topo I level present in 10 microg cell lysate of the human IGROV1 ovarian cancer cell line. We found no correlation between Topo I levels and (non-)hematological toxicity. Topo I levels after the fifth topotecan infusion were significantly negatively correlated with the AUC of topotecan (R = -0.64, p = 0.026), in contrast with Topo I levels prior to (R = -0.25, p = 0.4) and after (R = -0.30, p = 0.3) the first topotecan infusion. Topo I levels after the fifth topotecan infusion (48 +/- 27%, mean +/- SD) were higher than Topo I levels prior to and after the first topotecan infusion (3.0 +/- 4.7 and 2.7 +/- 3.6%, respectively) (p = 0.001). In conclusion, we detected a significant inverse correlation between Topo I level and topotecan AUC at day 5, and we found increasing Topo I levels during a daily x 5 schedule of treatment with topotecan. PMID- 11914647 TI - Population pharmacokinetics of high-dose etoposide in children receiving different conditioning regimens. AB - Pharmacokinetics after high-dose (HD) etoposide (Eto) (40 mg/kg i.v. once as 4-h infusion, one patient 20 mg/kg i.v. as 4-h infusion, for 3 consecutive days) were studied in 31 children and young adults (age 0.8-23.7 years, median: 8.0 years) undergoing bone marrow transplantation after different conditioning regimens. Blood samples were collected until 97 h after the end of infusion. The population analysis of the first part of data (112 samples/21 patients, well documented) served to establish the pharmacokinetic model. The same data combined with the second part of data (50 samples/10 patients, 'intention to treat') then served to calculate the final population model. Data were best described by a three compartment model with t1/2alpha = 0.28 h +/- 3.2%, t1/2beta = 3.6 h +/- 16.9% and t1/2gamma = 44.2 h +/- 56.5%, respectively (mean(geom) +/- CV(geom)). Clearance (CL) was 15.5 ml/min/m2 +/- 30.6% (mean(geom) +/- CV(geom)) and thus at the lower range of data reported in the literature. The fraction of unbound Eto (fu) was 7.0% (4.3-11.9%) [median (range)], with high intra-individual variability. An increase in f(u) with increasing total Eto was observed. The question of a principally lower Eto CL in children, as compared to adults, after HD treatment remains open. PMID- 11914646 TI - Antitumor activity of protein kinase C inhibitors and cisplatin in human head and neck squamous cell carcinoma lines. AB - Protein kinase C (PKC) plays a pivotal role in signal transduction involved in the control of cell proliferation, differentiation and apoptosis. Interference with such signaling pathways may result in altered tumor cell response to antineoplastic drugs. We investigated the effects of two selective PKC inhibitors as single agents and in combination with cisplatin in cell lines derived from squamous cell carcinomas of the head and neck (SCCHN). Safingol (Saf) is directed against the regulatory domain, whereas chelerythrine (Che) interacts with the catalytic domain of PKC. In six SCCHN cell lines (UM-SCC 11B, 14A, 14C and 22B, 8029NA, and a 5-fold cisplatin-resistant subline 8029DDP). PKC activities ranged between 1 and 158 IU/1 x 10(7) cells, and they were inversely proportional to the amount of cellular epidermal growth factor receptor. Using the colorimetric MTT assay, PKC inhibitors Saf and Che showed comparable dose-dependent growth inhibition. The 50% inhibitory concentrations (IC50) were between 3.8-8.6 microM for Saf and 8.5-13.6 microM for Che with no relationship to PKC activity or cisplatin sensitivity of the respective cell lines. Combinations of cisplatin (IC50 = 0.4-5.8 microg/ml) and either PKC inhibitor (5 microM Saf, 10 microM Che) led to a significant decrease of cisplatin IC50 values in most cell lines. However, comparison with theoretical additive dose-response curves showed additive rather than synergistic effects for both PKC inhibitors. PMID- 11914648 TI - NOR-1: a nitric oxide releasing agent for calibrating low levels of nitric oxide by the chemiluminescence method. AB - Nitric oxide (NO) is formed in small amounts in vivo and is rapidly oxidized by interacting with oxygen, making measurement of its level difficult. The chemiluminescence assay is the most widely used method for detecting NO and is extremely sensitive to very small amounts of NO. However, it is difficult to prepare small amounts of NO to be used as a standard for NO analysis. NOR-1, a derivative of NOR-3, is a newly discovered NO donor with rapid NO-releasing activity. We assessed the dynamics of NO release and decomposition using NOR-1. Our results demonstrate that NOR-1 is stable in dimethylsulfoxide (DMSO) and is able to dilute at lower concentration (to picomolar levels) by DMSO without decomposition. NOR-1 released persistently 1.4 more excess of NO with 15 min of incubation. There was a linear relationship between the concentration of NOR-1 and that of NO released from NOR-1 (r=0.997) These findings suggest that NOR-1 is a useful reagent for the calibration of lower NO detection. PMID- 11914649 TI - Prospective cross-sectional study of haemostatic factors in patients with and without coronary artery disease. AB - The role of haemostatic factors for arterial thrombosis, especially the prevalence of activated protein C (APC) resistance in patients with coronary artery disease (CAD), is controversial. Between November 1996 and August 1997, 665 patients were analyzed. Diagnosis of CAD was confirmed by coronary angiography, exclusion of CAD was accepted in the presence of negative stress testing or a negative coronary angiography. CAD was present in 370 (56%) and excluded in 295 (44%) patients. Patients with CAD were older (64 +/- 9.2 versus 57.7 +/- 16 years; P A mutation in the factor V gene. AB - A point mutation, 1691 G --> A in the coagulation factor V gene results in an Arg506 --> Gln amino acid substitution in the factor V molecule. This mutation, defined as factor VLEIDEN, results in activated protein C (APC) resistance and is the most common genetic risk factor for familial thrombophilia. A new mini sequencing method using matrix-assisted laser desorption/ionization time-of flight (MALDI-TOF) mass spectrometry was developed for the screening of the 1691G --> A substitution in factor V. In this method, a fragment of genomic DNA containing the 1691st base is first amplified, followed by mini-sequencing in the presence of dGTP and ddATP, ddCTP, and ddTTP. In this manner, the primer is extended by one base from one allele and two bases from the other allele. The extended products are analyzed using MALDI-TOF mass spectrometry. The base at position 1691 is identified based on the number of nucleotides added. We have used this method to genotype 16 APC-resistant patients previously identified by conventional methods and 11 normal control samples. The genotypes of all samples were correctly identified. This method is accurate, fast, and potentially allows for simultaneous multiplex genotyping of a number of mutation sites associated with thrombophilia and clot formation. PMID- 11914654 TI - Novel anticoagulant activity of polybrene: inhibition of monocytic tissue factor hypercoagulation following bacterial endotoxin induction. AB - The enhanced extrinsic coagulation in response to inflammation could contribute to disseminated intravascular coagulation, often manifesting cardiovascular complications. The complex mechanism remains unclear. Nor is the effective anticoagulation well established. The search for arresting hypercoagulation is of antithrombotic relevance. The ability of polybrene (PB) to inhibit tissue factor (TF)-initiated extrinsic blood coagulation was demonstrated at the protein and cellular levels as well as in human plasma samples. In a single-stage clotting assay, PB dose-dependently offset bacterial endotoxin (lipopolysaccharide) induced monocytic TF (mTF) hypercoagulation and inhibited rabbit brain thromboplastin (rbTF) procoagulation. Consistent with these findings, the significantly prolonged prothrombin time indicated the depressed extrinsic coagulation by PB. However, PB showed no effect on thrombin time. We dissected the extrinsic pathway to further determine the inhibitory site(s) of PB. A two stage chromogenic assay monitoring S-2288 hydrolysis showed that PB readily blocked mTF-dependent or rbTF-dependent FVII activation, which was verified by the diminished activated factor VII (FVIIa) formation derived from the proteolytic cleavage of its zymogen factor VII on Western blotting analyses. PB had no effect on FVIIa and activated factor X amidolytic activity. Nor was the dissected TF/FVIIa-catalyzed factor X activation affected. In conclusion, the preferential downregulation of factor VII activation was responsible for the depressed extrinsic coagulation. PB could present a novel anticoagulant antagonizing the extrinsic hypercoagulation for the prevention of thrombotic complication following sepsis and inflammations. PMID- 11914656 TI - Morphological and biochemical evidence of blood vessel damage and fibrinogenolysis triggered by brown spider venom. AB - The venom of the brown spider is remarkable because it causes dermonecrotic injury, hemorrhagic problems, hemolysis, platelet aggregation and renal failure. The mechanism by which the venom causes hemorrhagic disorders is poorly understood. Rabbits intradermally exposed to the venom showed a local hemorrhage starting 1 h after inoculation and reaching maximum activity between 2 and 3 days. Biopsies examined by light and transmission electron microscopy showed subendothelial blebs, vacuoles and endothelial cell membrane degeneration in blood vessels, plasma exudation into connective tissue, and fibrin and thrombus formation within blood vessels. Loxosceles intermedia venom incubated with fibrinogen partially degrades Aalpha and Bbeta chains of intact fibrinogen, and significantly cleaves all Aalpha, Bbeta and gamma chains when they were separated or when fibrinogen is denatured by boiling. Proteolytic kinetic studies showed that the Aalpha chain is more susceptible to venom hydrolysis than the Bbeta chain. The fibrinogenolysis is blocked by ethylenediamine tetraacetic acid and 1,10-phenanthroline, but not by other protease inhibitors. Human plasma incubated with the venom had coagulation parameters such as prothrombin time, activated partial thromboplastin time and thrombin time increased. Through molecular sieve chromatography, we isolated a venom toxin of 30 kDa with fibrinogenolytic activity. We propose that the local and systemic hemorrhagic disorders evoked in loxoscelism are consequences of direct venom fibrinogenolysis together with cytotoxicity to subendothelial structures and endothelial cells in blood vessels. PMID- 11914655 TI - Evaluation of the blood coagulation mechanism and platelet aggregation in individuals with mechanical or biological heart prostheses. AB - Oral anticoagulants have been widely employed to decrease thrombotic risk by reducing the levels of vitamin K-dependent clotting factors. Paradoxically, the use of oral anticoagulants also decreases the levels of natural anticoagulants (protein C and protein S), which favors the hypercoagulability state. Increased platelet activation has been reported in patients undergoing warfarin treatment. These findings have raised questions about the antagonistic effect of oral anticoagulants and their implications for hemostatic balance. The aim of this study is to determine the relationship between warfarin dosage and prothrombin time [International Normalized Ratio (INR)], platelet aggregation, vitamin K dependent clotting factors, and protein C and protein S. Blood samples from 27 patients were analyzed, seven with mechanical prostheses and 20 with biological prostheses, and 27 controls. Multiple regression analysis showed that factor II most significantly determines the INR. Results showed that the INR, clotting factors, and protein C and protein S activity did not correlate with warfarin dosage, highlighting the need for accurate laboratory monitoring of those undergoing anticoagulant therapy. PMID- 11914657 TI - Fibrinogen Saint-Germain I: a case of the heterozygous Aalpha GLY 12 --> VAL fibrinogen variant. AB - A fibrinogen variant was suspected based on the results of routine coagulation tests in a 2-year-old asymptomatic child. Coagulation studies showed marked prolongation of both the thrombin and reptilase times, and discrepancy was noted between the level of plasma fibrinogen as measured by a kinetic versus immunological determination. Family studies revealed that the father beared the same abnormality. Studies of purified fibrinogen revealed an impaired release of both fibrinopeptides by thrombin. Fibrin monomer polymerization and fibrin stabilization were normal. DNA sequencing revealed a heterozygous G --> T point mutation in exon 2 of the gene coding for the Aalpha chain, which substituted a Gly for Val at position 12. Although the mutation is the same as in fibrinogen Rouen, fibrinogen Saint-Germain I shows a different fibrinopeptide release pattern and a mild factor V deficiency. PMID- 11914658 TI - An evaluation of screening tests for defects in the protein C pathway: commercial kits lack sensitivity and specificity. AB - A comparative evaluation of four commercial coagulation test kits for screening the protein C pathway kits was performed at two centres. The tests were Acticlot V-OUT (V-OUT), the PCA test (PCA), the GradiThrom PCP test (PCP) and ProC Global (ProC). Reference ranges were established in 40 normal subjects and, with the exception of V-OUT and ProC, significant differences were observed between males and females. Consequently, sex-specific normal cut-off values (fifth percentile) were used that led to greatly improved sensitivity when compared with the manufacturers' recommended cut-off values. Plasma from patients with factor V Leiden (n = 23), congenital protein S deficiency (n = 19), congenital protein C deficiency (n = 11), lupus anticoagulant (n = 17) and thrombophilia with no demonstrable protein C pathway defect (n = 20) were tested. All kits showed 100% sensitivity to factor V Leiden, but sensitivity was variable for protein C deficiency (27-73%), and poor for protein S deficiency (29-35%). The lupus anticoagulant affected all kits to some degree, with 29-35% giving values below the fifth percentile of normal, whereas all kits gave 1/20 unexpected abnormal results in the thrombophilia group, with the same sample accounting for the abnormal results in three of the four kits. Overall sensitivity and specificity, respectively, for defects in the protein C pathway were: V-OUT, 60 and 91%; PCA, 70 and 86%; PCP 75 and 94%; and ProC, 66 and 88%. We conclude that all four kits lack the sensitivity and specificity required for routine laboratory screening for defects in the protein C pathway and cannot replace assays for the individual proteins of this system. PMID- 11914659 TI - Increased soluble intercellular adhesion molecule-1, breast cancer and the acute phase response. AB - Abnormal levels of soluble intercellular adhesion molecule-1 (sICAM-1), C reactive protein (CRP) and von Willebrand factor (vWf) are commonly found in breast and other cancers. However, the relationship between these molecules is unclear as raised sICAM-1 and vWf may simply reflect an acute phase response. We tested the hypothesis that raised sICAM-1 and vWf are related to the acute phase response by measuring both markers by enzyme-linked immunosorbent assay, and CRP by dry chemistry, in 40 women with breast cancer (cases) and 40 age-matched controls. All three markers were raised in the cases compared with the controls [sICAM-1 (mean +/- SD), 281 +/- 70 versus 230 +/- 40 ng/ml; vWf (mean +/- SD), 139 +/- 33 versus 106 +/- 16 IU/dl; CRP (median), 7 (interquartile range, 5-11) versus 5 (4-6) mg/dl; all P < 0.01]. However, sICAM-1 correlated strongly with CRP (r = 0.67, P < 0.001) in the cases but there was no significant relationship between vWf and CRP (r = -0.15, P = 0.351). There were no significant correlations in the controls. We conclude that raised sICAM-1 in breast cancer, like the acute phase response, reflects inflammation, and that raised vWf seems likely to be independent of the acute phase response. PMID- 11914660 TI - Hyperhomocysteinemia is common in patients with antiphospholipid syndrome and may contribute to expression of major thrombotic events. AB - Homocysteine plasma levels were measured by high-performance liquid chromatography in 52 patients with primary antiphospholipid syndrome (APS). Elevated homocysteine concentrations (>or= 15 micromol/l) were found in 16/52 (30.8%) APS patients. Elevated homocysteine levels were found to be associated with an increased risk for a major thromboembolic event (50 versus 16.7%, P = 0.014). Hyperhomocysteinemia is a common finding in APS patients, and may contribute to severity of thrombotic tendency observed in this syndrome. PMID- 11914661 TI - The utility of Tc-99m dextran in the diagnosis and identification of melanoma metastases responsible for protein-losing enteropathy. AB - Protein-losing enteropathy is an uncommon syndrome of excessive loss of protein via the gastrointestinal mucosa. Tc-99m dextran is a tracer ideally suited for diagnosis and localization. The authors report a case of melanoma mestastases to the small bowel that were causing protein-losing enteropathy. These were diagnosed and localized using Tc-99m dextran, leading to a curative resection. PMID- 11914662 TI - The efficacy of sestamibi parathyroid scintigraphy for directing surgical approaches based on modified interpretation criteria. AB - PURPOSE: With the increasing use of targeted parathyroidectomy, the accuracy of sestamibi parathyroid scintigraphy (SPS) in determining the best surgical approach has become more clinically relevant than its sensitivity for detecting all abnormal glands. The reported accuracy of SPS does not represent the true efficacy of SPS for directing targeted parathyroidectomy, because many authors reported accuracy on the basis of abnormal glands. The authors assessed the efficacy of SPS based on modified interpretation criteria to determine specifically whether SPS influences the surgeon's choice of a targeted versus a conventional approach to parathyroidectomy. METHODS: The authors reviewed the SPS studies (both dual-isotope subtraction and Tc-99m sestamibi dual-phase techniques) performed in 80 patients with primary hyperparathyroidism. All patients had surgery with histologic diagnoses, intraoperative PTH assays, and clinical follow-up. RESULTS: Of 75 patients with solitary adenomas, 9 SPS studies were negative, equivocal, or showed two or more foci, and 66 studies showed a solitary focus on the correct side. Of these 66 studies, 63 showed a lesion in the correct quadrant (e.g., superior or inferior) and 3 predicted the correct side only but a wrong quadrant. The positive-predictive value of SPS was 100% for correctly identifying the side of the adenoma (thus correctly directing unilateral surgery) and 95.5% for correctly identifying the quadrant of the solitary adenoma. CONCLUSION: The positive-predictive value of SPS for directing the surgical approach (but not for detecting individual lesions) that reveals a single focus is very high based on our modified interpretation criteria. PMID- 11914663 TI - Presurgical localization of abnormal parathyroid glands using a single injection of Tc-99m sestamibi: comparison of high-resolution parallel-hole and pinhole collimators, and interobserver and intraobserver variation. AB - PURPOSE: Scintigraphy using Tc-99m sestamibi is sensitive in localizing abnormal parathyroid glands in patients with hyperparathyroid disease (HPT). Various methods have been described to increase the sensitivity of the single-tracer technique, such as SPECT, factor analysis of dynamic structures, and use of a pinhole collimator, but often the gain in sensitivity is accompanied by a loss of specificity. PURPOSE: In this study, the authors compared the sensitivity and specificity of side localization of the diseased gland(s) using the double-phase single-tracer method performed with and without the addition of a pinhole collimator in the early and late phases of imaging. The combined high-resolution parallel-hole and pinhole collimator imaging protocol was further validated by investigation of interobserver and intraobserver variation. MATERIALS AND METHODS: Forty-seven patients with primary HPT and 16 patients with secondary HPT examined from 1996 to 1999 with the Tc-99m sestamibi double-phase technique and who had subsequent surgery formed the basis of the study. Their histologic and follow-up data were also factored into this analysis. Tc-99m sestamibi (750 to 900 MBq; 18.9 to 24.3 mCi) was injected. Ten-minute neck and mediastinum images acquiring 1,000 K counts were obtained with the high-resolution parallel-hole collimator, and a neck image containing 500 K counts was obtained with the pinhole collimator. Two to three hours later, the same parallel-hole and pinhole collimator images were obtained that had the same acquisition time as the early images. Two observers who were nuclear medicine specialists independently viewed all the parallel-hole scintigrams and afterward all parallel-hole and pinhole scintigrams two times. RESULTS: Thirty-eight (81%) of the patients with primary HPT had a single adenoma. The sensitivity and specificity for the correct side of localization were 54% and 89%, respectively, using the high-resolution parallel hole collimator, and 88% and 77%, respectively, with the addition of the pinhole collimator in all patients with primary HPT. In patients with secondary HPT, the sensitivity and specificity for localization of the correct side were 58% and 100%, respectively, using the high-resolution parallel-hole collimator, and 85% and 100%, respectively, with the addition of the pinhole collimator. The interobserver agreement was acceptable, with an overall agreement of 84% and a kappa value of 0.67. The intraobserver agreement was even better, with an overall agreement of 88% and 90% and kappa values of 0.76 and 0.79 for the two observers. CONCLUSIONS: Sensitivity is increased considerably when the pinhole collimator is added to the imaging protocol of parathyroid scintigraphy in patients with primary or secondary HPT. A loss of specificity occurred only in patients with primary HPT. The precision of the combined approach is very high. PMID- 11914664 TI - Increased prognostic value of combined myocardial perfusion SPECT imaging and the quantification of lung Tl-201 uptake. AB - PURPOSE: The authors wanted to identify those patients assessed by exercise SPECT in whom the quantification of lung Tl-201 uptake helps to evaluate disease prognosis. METHODS: One hundred forty-nine patients (114 men, 35 women; 74 after myocardial infarction [MI]; mean age, 54 +/- 9 years) underwent exercise Tl-201 SPECT. The SPECT patterns were divided into normal (n = 45), fixed defects (n = 29), and inducible ischemia (n = 75). Anterior planar imaging was performed before SPECT acquisition to calculate the lung-to-heart ratio (L:H). RESULTS: During an average follow-up of 20 +/- 9 months, eight patients had died of cardiac causes and 13 patients experienced nonfatal MIs. Among the 45 patients with normal perfusion, no cardiac event was observed and the L:H ratio was not helpful for risk stratification. In 29 patients with fixed defects, four cardiac deaths occurred (all in patients with L:H ratios >0.5; annual event rate, 21.1% for L:H ratios >0.5 compared with 0% for L:H ratios <0.5; chi-square = 4.07, P < 0.05). Among the 75 patients with ischemia, 4 died and 13 had nonfatal MIs (annual event rate, 15.4% for L:H ratios >0.5 compared with 13% for L:H ratios <0.5; P = NS). CONCLUSIONS: These findings suggest a benign prognosis in patients with normal SPECT (regardless of the L:H ratio). Conversely, all patients with ischemia are at high risk for future cardiac events. Quantification of the Tl-201 lung uptake seems to be valuable in evaluations of disease prognosis, especially in patients with fixed defects. PMID- 11914665 TI - Massive hemangioma or lymphedema? A case with a diagnostic dilemma. AB - The authors present an unusual case of massive enlargement of the right lower extremity that was treated as lymphedema secondary to filarial infection. A multiple-method approach beginning with scintigraphy led to the diagnosis of massive hemangioma with probable associated lymphatic proliferation. Abnormal local proliferation of vascular and lymphatic tissue is a developmental anomaly that is difficult to diagnose or characterize, even with histopathologic data. Individual imaging methods may also yield nonspecific findings. Blood-pool scintigraphy, therefore, has a definitive role to play in assessing such complex lesions. PMID- 11914667 TI - Multiple hepatic adenomas: Tc-99m RBC liver SPECT findings with pathologic correlation. AB - A 24-year-old man with multiple hepatic masses incidentally detected by ultrasonography underwent Tc-99m RBC liver SPECT. SPECT images showed increased blood pooling mimicking hemangiomas in two of four masses or activity similar to normal liver in the others. The patient had a 3-year history of steroid therapy for rheumatoid arthritis. Computed tomography and magnetic resonance imaging showed multiple liver lesions. Angiography revealed three hypervascular and one hypovascular mass. Microscopic examination of the enucleated specimens disclosed hepatic adenomas with many dilated sinusoids in the masses showing blood pooling and fatty change, and adenomas without dilated sinusoids in the masses not showing blood pooling. This case report shows that the appearance of hepatic adenomas can vary on Tc-99m RBC liver SPECT, depending on whether dilated sinusoid and hepatic adenomas show blood pooling. This is one of the few examples of false-positive findings that can be mistaken for hemangioma. PMID- 11914666 TI - Decreased thyroid uptake of Tc-99m pertechnetate in patients with advanced-stage Sjogren syndrome: evaluation using salivary gland scintigraphy. AB - PURPOSE: The authors assessed the uptake of Tc-99m pertechnetate in the thyroid using salivary gland scintigraphy in patients with Sjogren syndrome and in healthy controls. MATERIALS AND METHODS: Salivary gland scintigraphy and a labial biopsy were performed in 73 patients with Sjogren syndrome. Based on the labial biopsy findings, 32 patients with a histopathologic grade of 1 or 2 were regarded as having early-stage Sjogren syndrome and 41 patients with a grade of 3 or 4 were regarded as having an advanced stage. After the administration of 370 MBq (10 mCi) Tc-99m pertechnetate, dynamic salivary gland scintigraphy was performed for 50 minutes. Lemon juice was used to stimulate the salivary glands, and the thyroid gland was included in the imaging area. Scintigraphy was also performed in an age- and sex-matched control group of 25 healthy persons. The thyroid uptake ratio was calculated for the scintigraphic images and compared among the three groups: healthy controls, patients with early-stage Sjogren syndrome, and those with advanced-stage Sjogren syndrome. RESULTS: When compared with the control group, the thyroid uptake ratio of the early-stage Sjogren syndrome group was not significantly different, whereas that of the advanced-stage group was significantly lower. CONCLUSIONS: Thyroid uptake of Tc-99m pertechnetate was less in patients with advanced-stage Sjogren syndrome than in patients with early stage Sjogren syndrome or in healthy controls. Measuring the thyroid uptake of Tc 99m pertechnetate using salivary gland scintigraphy is an easy and useful method for assessing thyroid disorders in Sjogren syndrome and thus should be performed routinely. PMID- 11914668 TI - Tumor localization and image registration of F-18 FDG coincidence detection scans with computed tomographic scans. AB - PURPOSE: The aim of this study was to determine the feasibility of registering routine clinical F-18 fluorodeoxyglucose (FDG) coincidence detection (CD) scans with computed tomographic (CT) scans for radiation treatment planning and case management. METHODS: F-18 FDG CD and chest CT scans, performed in 10 randomly selected patients with confirmed or possible adenocarcinoma of the lung, were evaluated. The quality of the matches was verified by comparisons of the center to-center distance between a region of interest (ROI) manually drawn on the CT slice and warped onto the CD slice with an ROI drawn manually directly on the CD slice. In addition, the overlap between the two ROIs was calculated. RESULTS: All 10 F-18 FDG CD and CT scans were registered with good superimposition of soft tissue density on increased radionuclide activity. The center-to-center distance between the ROIs ranged from 0.29 mm to 8.08 mm, with an average center-to-center distance of 3.89 mm +/- 2.42 mm (0.69 pixels +/- 0.34 pixels). The ROI overlap ranged from 77% to 99%, with an average of 90% +/- 5.6%. CONCLUSIONS: Although the use of F-18 FDG CD shows great promise for the identification of tumors, it shares the same drawbacks as those associated with radiolabeled monoclonal antibody SPECT and ligand-based positron emission tomographic scans in that anatomic markers are limited. This study shows that image registration is feasible and may improve the clinical relevance of CD images. PMID- 11914669 TI - Incidental finding of upper extremity deep venous thrombosis on skeletal scintigraphy. PMID- 11914670 TI - Chest tube insertion as a potential source of false-positive FDG-positron emission tomographic results. PMID- 11914672 TI - Vesicoureteral reflux causing misinterpretation of a Tc-99m DTPA scan. PMID- 11914671 TI - Urinary diarrhea detected by Tc-99m DTPA scintigraphy in a 3-year-old girl. PMID- 11914673 TI - Parastomal hemorrhage demonstrated on a red blood cell-labeled gastrointestinal bleeding site study. PMID- 11914674 TI - SPECT demonstrating lymphatic drainage from truncal melanoma to para-aortic lymph nodes. PMID- 11914675 TI - Tc-99m MDP and Tc-99m MIBI in the assessment of spondyloarthritis presenting as bone metastasis before treatment with infliximab. PMID- 11914676 TI - Unusual bilateral symmetrical osteolytic metastases visualized by bone scintigraphy. PMID- 11914677 TI - A large pheochromocytoma seen on dynamic bone scintigraphy. PMID- 11914678 TI - Harness-shaped distribution in pediatric Tc-99m tetrofosmin scintigraphy. PMID- 11914679 TI - Parathyroid adenoma visualization on octreotide scintigraphy. PMID- 11914680 TI - Accumulation of Tc-99m tetrofosmin in multiple myeloma. PMID- 11914681 TI - Hepatic abscess detected on a three-phase bone scan. PMID- 11914682 TI - A hypophyseal metastasis of follicular thyroid carcinoma. PMID- 11914683 TI - Bone imaging of sternal metastasis from hepatocellular carcinoma with disease progression. PMID- 11914684 TI - Current readings in nuclear medicine. PMID- 11914686 TI - Preterm labour. AB - Studies in the past year have clarified the roles of inflammatory mediators in preterm labour. Exploration of possible genetic predisposition is just beginning. Ultrasound measurement of cervical length has the potential to predict women at risk of preterm delivery several weeks before it occurs. Biochemical testing such as fetal fibronectin can possibly increase its predictive value and differentiate true preterm labour from more innocent preterm contractions. The use of antibiotics for preterm premature rupture of membranes has been clarified with the ORACLE I trial, which shows health benefits for the neonate with the use of erythromycin, whereas antibiotics do not seem to play a beneficial role in spontaneous preterm labour without evidence of clinical infection. There have been further studies suggesting that agents other than beta-agonists are preferable for acute tocolysis and that repeated doses of corticosteroids should be used with caution. PMID- 11914687 TI - Bacterial vaginosis in pregnancy. AB - Bacterial vaginosis is associated with adverse pregnancy outcomes across all gestational ages. It is linked to first and second trimester fetal loss, chorioamnionitis, preterm delivery, low-birthweight infants and maternal/neonatal infectious morbidity. Infants who survive preterm birth are at an increased risk of subsequent neurodevelopmental delays and handicap, more so if there was underlying choriomnionitis. The exact mechanisms and pathways through which bacterial vaginosis exerts these adverse effects are incompletely understood. Not surprisingly, intervention studies in bacterial vaginosis-positive pregnant women have shown conflicting results, both in women at high and low risk of preterm birth. A much better understanding of the pathobiology of bacterial vaginosis in pregnancy is required to focus the designs of intervention studies on the disturbed cellular and biochemical pathways. Such studies may address the benefits of the treatment of bacterial vaginosis before conception and during early pregnancy, to determine whether treatment in populations of pregnant women may be beneficial. PMID- 11914688 TI - Hypertensive diseases and eclampsia. AB - Worldwide, pre-eclampsia and eclampsia contribute to the death of a pregnant woman every 3 min. In the UK in recent decades, hypertensive disorders of pregnancy have remained one of the leading causes of both maternal and perinatal morbidity and mortality. The management of pregnancies complicated by hypertension has not significantly altered for many years, possibly as a result of little progress being made in our understanding of the condition. New insights, however, have recently been gained into the pathophysiology of pre eclampsia. These have yet to be translated into new interventions or to make any impact on clinical management of these pregnancies. This review will therefore focus on recent advances relating to research into the aetiology and pathogenesis of pre-eclampsia, but will conclude with a brief update on current therapeutic strategies. PMID- 11914689 TI - Intrauterine growth restriction. AB - Pathophysiological processes underlying intrauterine growth restriction are very complex and poorly understood. Growth restricted fetuses are at risk of hypoxia and, therefore, an early diagnosis of intrauterine growth restriction is important for initiation of fetal surveillance. Application of a three dimensional ultrasound method for estimation of fetal weight promises better precision. Use of conditional standard deviation scores of fetal biometric variables has been suggested for improved individualized evaluation of intrauterine growth. Application of umbilical artery Doppler velocimetry in the clinical management of growth-restricted fetuses after 32 weeks of gestation leads to decreased perinatal mortality and lower rates of obstetric interventions. Evaluation of fetal state before 32 weeks is difficult and should include Doppler examination of placental circulation and several fetal arterial and venous vessel beds. In addition, recordings of short-term variability of fetal heart rate and biophysical profile have been suggested for fetal surveillance. Important new data on the time sequence of Doppler changes in various vessels of compromised very preterm growth restricted fetuses have been presented, which will enable the establishment of clinical management protocols for evaluation in prospective randomized studies. PMID- 11914691 TI - Smoking in pregnancy. AB - This update discusses the evidence to link maternal smoking during pregnancy with adverse pregnancy outcome, and also the potential long-term effects on health of in-utero exposure to tobacco smoke. Smoking cessation strategies will also be considered. PMID- 11914690 TI - Cardiac disease in pregnancy. AB - Congenital heart disease in pregnancy is increasingly common because of the advances in surgery and medical therapy which have taken place over the last 30 years, which means that more affected women are surviving into the reproductive age. Antenatal counselling needs to be tailored to the specific lesion, with pulmonary hypertension and cyanotic disease presenting a risk of maternal mortality of up to 50%. The use of anticoagulants in women with artificial valves presents a particular challenge, heparin being safer for the baby and warfarin for the mother. Peripartum cardiomyopathy and Marfan's syndrome may be less dangerous than once thought. The risk of congenital heart disease in the fetus is increased, from twice to 20-fold, depending on the nature of the mother's lesion. Care throughout pregnancy and in the puerperium should be multidisciplinary and include cardiologists, obstetricians and midwives with experience of such cases, preferably in a tertiary centre. PMID- 11914692 TI - Recent advances in neonatology. AB - This update focuses on recent neonatal research of potential interest to obstetricians. Accurate information on outcomes for infants born at the edges of viability is critical to informing management decisions. New research, population based, gives guidance on short-term survival and long-term neurodevelopmental outcome. Recent research has also highlighted the variation in attitudes to end of-life decision-making and important influences on this that cross different boundaries. Although research is dominated by issues related to prematurity, some important developments relevant to term infants will be covered. There is accumulating evidence that challenges the traditional approach of using 100% O2 in resuscitation. For infants suffering intrapartum asphyxia there are new approaches to neuroprotection actively being explored. Therapeutic interventions such as extracorporeal membrane oxygenation and inhaled nitric oxide, available for some time, are having their place in neonatal intensive care more clearly defined. PMID- 11914694 TI - Developments in laboratory techniques for prenatal diagnosis. AB - Ongoing trends in prenatal diagnosis aim at early, rapid, and ideally noninvasive diagnosis as well as at the improvement of risk-screening for aneuploidy. Interphase-fluorescence in situ hybridization and quantitative fluorescence polymerase chain reaction are efficient tools for the rapid exclusion of selected aneuploidies in addition to the established direct preparation of chromosomes from chorionic villi. Interphase fluorescence in situ hybridization has also made possible the diagnosis of selected chromosome abnormalities in single cells (e.g. in preimplantation genetic diagnosis) or noninvasive diagnosis. More complex multicolor fluorescence in situ hybridization approaches are currently being evaluated. Single cell polymerase chain reaction is the key technique for the molecular diagnosis of a growing number of monogenic conditions before implantation or, still more experimental, in fetal cells retrieved from the maternal circulation. New sources for noninvasive diagnosis came into play such as fetal DNA or cell nuclei in maternal plasma. The combination of biochemical parameters in the maternal serum, namely free beta-human chorionic gonadotropin with pregnancy associated plasma protein A and sonographic markers, has already dramatically increased the sensitivity of risk screening in the first trimester of pregnancy. PMID- 11914695 TI - Prenatal diagnosis for multiple pregnancies. AB - In multiple pregnancies, first trimester ultrasound is crucial to diagnose chorionicity, to detect major structural defects, and to screen for chromosomal abnormalities based on nuchal translucency measurement. The efficacy of nuchal translucency measurement screening in twins might be improved when combined with first trimester maternal serum screening. In twins as in singletons, the risk of fetal loss attendant to chorionic villi sampling and to amniocentesis are similar. When an invasive procedure is indicated in twins, chorionic villi sampling has, over amniocentesis, the advantage of allowing selective termination to be performed in the first trimester, when the procedure related risk of miscarriage is minimal. It has the disadvantage of leading to ambiguous results in up to 2% of cases. While chorionic villi sampling is the choice technique in pregnancies at very high risk, amniocentesis is still indicated in cases at more moderate risk. In monochorionic pregnancies, selective termination can now be performed using a variety of techniques including bipolar or monopolar cord coagulation, and, in acardiac twins, alcohol ablation. However, selective termination remains more hazardous in monochorionic than in dichorionic pregnancies. The outcome of the twin-to-twin transfusion syndrome has been substantially improved by laser photocoagulation of placental shunts and by amniodrainage, but randomized trials are needed to establish the optimal therapeutic strategy, and further pathophysiologic research might result in new treatments. PMID- 11914696 TI - Prediction of fetal anaemia. AB - The perinatal outcome of the anaemic fetus has dramatically improved over the last 20 years, as a result of early recognition of the problem and treatment by intrauterine transfusion. Traditionally assessment of the anaemic fetus relied on obstetric history and maternal antibody titre, which proved to be inadequate tests to accurately predict fetal condition. More recently, invasive testing with techniques such as amniocentesis and cordocentesis have allowed a more accurate evaluation of the degree of anaemia, while at the same time enabling transfusion to take place. Such techniques are not without danger, with perinatal loss and fetomaternal haemorrhage being significant risks. The clinical community has therefore sought to find accurate, non-invasive methods for assessing the degree of fetal anaemia, thereby reducing the number of unnecessary invasive procedures, while at the same time providing more precise data on the quantity and timing of the transfusion. Recent publications focusing on the diagnosis and management (including plasmapheresis, immunoglobulins and intrauterine transfusion) of fetal anaemia will be discussed. PMID- 11914697 TI - Prenatal evaluation for fetal surgery. AB - The selection of fetuses that may benefit with in utero surgery is being developed. Noninvasive and invasive prenatal diagnosis techniques are utilized to try and gain as much knowledge about the fetus so that the appropriate counseling of parents can be undertaken. The most common invasive techniques are amniocentesis and chorionic villus sampling for fetal karyotyping and genetic diagnosis. Noninvasive techniques include ultrasound (2D and 3D), fetal echocardiography and magnetic resonance imaging. Additional techniques such as specific Doppler evaluation of vascular components, new techniques to look at fetal electrocardiograms and the use of computer tomography are also considered. The most common conditions for which in utero fetal surgery is also being considered are twin to twin transfusion syndrome, myelomeningocele, sacrococcygeal teratoma, cystic adenomatoid malformation of the lung with fetal hydrops and other monochorionic twin abnormalities (severe discordant birth defects or twin reversal arterial perfusion sequences). Ongoing evaluation of the sensitivity, specificity, positive and negative predicted values of these evaluation tools is required so that appropriate selection of fetuses for the surgery can be made. PMID- 11914698 TI - Cervical screening for preterm delivery. AB - The ability of sonographic cervical length screening to detect those at risk of spontaneous preterm delivery has been extensively explored over the past few years. This applies both to high-risk and low-risk groups. Cervical length measurement appears to be superior to biochemical, microbiological or hormonal methods of screening. The screening technique has been standardized, but the cervical length for intervention and the timing and nature of intervention have not been defined. Cervical cerclage appears to be of use to prevent or arrest the progress of cervical dilation in high-risk cases, but the management of the screen-positive low-risk case has yet to be determined. Future management may be stratified according to actual cervical length, and prospective randomized trials of treatment for the short cervix are needed. PMID- 11914699 TI - Prolonged pregnancy. AB - Prolonged pregnancy is defined as any pregnancy that lasts 294 days or more. It is now well recognized that prolonged pregnancy is associated with an increased risk of perinatal mortality and morbidity. It is these complications of pregnancy that have led obstetricians to adopt a policy of induction of labour before the onset of the post-term period. The induction of labour between 41 and 42 weeks is, however, a very crude strategy for reducing term and post-term stillbirth rates. Although the risk of fetal death is increased after 42 weeks, many more fetuses die in utero between 37 and 42 weeks than die in the post-term period. It appears that smaller term fetuses run a greater risk than their larger counterparts, and that current methods of antepartum assessment of the term fetus are still inadequate. It behoves us as obstetricians to improve our capabilities in identifying the compromised fetus at term. This review puts into perspective the most recent publications and highlights areas requiring further study. PMID- 11914700 TI - Anatomy and examination of the eyelids. PMID- 11914701 TI - Ptosis in infants and children. PMID- 11914702 TI - Ptosis in young adults. PMID- 11914703 TI - Upper and lower eyelid retraction. PMID- 11914704 TI - Ptosis in the elderly patient. PMID- 11914705 TI - Distichiasis, trichiasis, and entropion: advances in management. PMID- 11914706 TI - Dermatochalasis. PMID- 11914707 TI - Facial nerve paralysis. PMID- 11914708 TI - Benign essential blepharospasm. PMID- 11914709 TI - Traumatic eyelid injuries. PMID- 11914710 TI - Benign eyelid tumors and skin diseases. PMID- 11914711 TI - Advances in the management of malignant tumors of the eyelid and conjunctiva: the role of sentinel lymph node biopsy. PMID- 11914713 TI - Activists threaten British neuroscience. PMID- 11914714 TI - A proposed mechanism of ALS fails the test in vivo. PMID- 11914715 TI - Flies put the buzz back into long-term-potentiation. PMID- 11914716 TI - A broad view of glutamate spillover. PMID- 11914717 TI - A new window on sound. PMID- 11914718 TI - Color visions in the brain. PMID- 11914719 TI - Protein kinase Mzeta is necessary and sufficient for LTP maintenance. PMID- 11914721 TI - Retinal ganglion cell synchronization by fixational eye movements improves feature estimation. AB - Image movements relative to the retina are essential for the visual perception of stationary objects during fixation. Here we have measured fixational eye and head movements of the turtle, and determined their effects on the activity of retinal ganglion cells by simulating the movements on the isolated retina. We show that ganglion cells respond mainly to components of periodic eye movement that have amplitudes of roughly the diameter of a photoreceptor. Drift or small head movements have little effect. Driven cells that are located along contrast borders are synchronized, which reliably signals a preceding movement. In an artificial neural network, the estimation of spatial frequencies for various square wave gratings improves when timelocked to this synchronization. This could potentially improve stimulus feature estimation by the brain. PMID- 11914720 TI - Memory enhancement and formation by atypical PKM activity in Drosophila melanogaster. AB - Synaptic stimulation activates signal transduction pathways, producing persistently active protein kinases. PKMzeta is a truncated, persistently active isoform of atypical protein kinase C-zeta (aPKCzeta), which lacks the N-terminal pseudosubstrate regulatory domain. Using a Pavlovian olfactory learning task in Drosophila, we found that induction of the mouse aPKMzeta (MaPKMzeta) transgene enhanced memory. The enhancement required persistent kinase activity and was temporally specific, with optimal induction at 30 minutes after training. Induction also enhanced memory after massed training and corrected the memory defect of radish mutants, but did not improve memory produced by spaced training. The 'M' isoform of the Drosophila homolog of MaPKCzeta (DaPKM) was present and active in fly heads. Chelerythrine, an inhibitor of PKMzeta, and the induction of a dominant-negative MaPKMzeta transgene inhibited memory without affecting learning. Finally, induction of DaPKM after training also enhanced memory. These results show that atypical PKM is sufficient to enhance memory in Drosophila and suggest that it is necessary for normal memory maintenance. PMID- 11914722 TI - Reorganization of human cortical maps caused by inherited photoreceptor abnormalities. AB - We describe a compelling demonstration of large-scale developmental reorganization in the human visual pathways. The developmental reorganization was observed in rod monochromats, a rare group of congenitally colorblind individuals who virtually lack cone photoreceptor function. Normal controls had a cortical region, spanning several square centimeters, that responded to signals initiated in the all-cone foveola but was inactive under rod viewing conditions; in rod monochromats this cortical region responded powerfully to rod-initiated signals. The measurements trace a causal pathway that begins with a genetic anomaly that directly influences sensory cells and ultimately results in a substantial central reorganization. PMID- 11914723 TI - Functional magnetic resonance imaging of synesthesia: activation of V4/V8 by spoken words. AB - In 'colored-hearing' synesthesia, individuals report color experiences when they hear spoken words. If the synesthetic color experience resembles that of normal color perception, one would predict activation of parts of the visual system specialized for such perception, namely the human 'color center', referred to as either V4 or V8. Using functional magnetic resonance imaging (fMRI), we here locate the region activated by speech in synesthetes to area V4/V8 in the left hemisphere, and demonstrate overlap with V4/V8 activation in normal controls in response to color. No activity was detected in areas V1 or V2, suggesting that activity in primary visual cortex is not necessary for such experience. Control subjects showed no activity in V4/V8 when imagining colors in response to spoken words, despite overtraining on word-color associations similar to those spontaneously reported by synesthetes. PMID- 11914724 TI - Callosotomy patients exhibit temporal uncoupling during continuous bimanual movements. AB - Rhythmic bimanual movements are highly constrained in the temporal domain, with the gestures of the two hands tightly synchronized. Previous studies have implicated a subcortical locus for temporal coupling based on the observation that these constraints persist in callosotomy patients. We now report that such coupling is restricted to movements entailing a discrete event (such as a movement onset). Three callosotomy patients exhibited a striking lack of temporal coupling during continuous movements, with the two hands oscillating at non identical frequencies. We propose a subcortical locus of temporal coupling for movements involving discrete events. In contrast, synchronization between the hands during continuous movements depends on interhemispheric transmission across the corpus callosum. PMID- 11914725 TI - Advanced online publication. PMID- 11914726 TI - Powering mitochondrial protein import. PMID- 11914727 TI - Effector regulation in a monomeric enzyme. PMID- 11914728 TI - How calpain is activated by calcium. PMID- 11914729 TI - Single-handed recognition of a sorting traffic motif by the GGA proteins. PMID- 11914730 TI - Picture story. Does Dengue virus fuse using beta-barrels? PMID- 11914731 TI - Max Ferdinand Perutz OM FRS. PMID- 11914735 TI - Mice with targeted gene disruptions or gene insertions for diabetes research: problems, pitfalls, and potential solutions. AB - The mouse has been a favoured organism for molecular manipulation in studies seeking to establish the genetic bases and pathophysiologic mechanisms underlying both Type I (insulin-dependent) and Type II (non-insulin-dependent) diabetes mellitus. Gene targeting and transgenesis are the two powerful molecular technologies used in these endeavours. Interpretation of results generated from such studies, either entailing an altered phenotype or the absence of a phenotypic change, is not always simple. This review focuses on certain complications inherent in the methodologies, and outlines steps that can be taken to distinguish effects of the genetic manipulation from unexpected contributions from the genetic background. PMID- 11914736 TI - Regulation of insulin gene transcription. AB - The mammalian insulin gene is exclusively expressed in the beta cells of the endocrine pancreas. Two decades of intensive physiological and biochemical studies have led to the identification of regulatory sequence motifs along the insulin promoter and to the isolation of transcription factors which interact to activate gene transcription. The majority of the islet-restricted (BETA2, PDX-1, RIP3b1-Act/C1) and ubiquitous (E2A, HEB) insulin-binding proteins have been characterized. Transcriptional regulation results not only from specific combinations of these activators through DNA-protein and protein-protein interactions, but also from their relative nuclear concentrations, generating a cooperativity and transcriptional synergism unique to the insulin gene. Their DNA binding activity and their transactivating potency can be modified in response to nutrients (glucose, NEFA) or hormonal stimuli (insulin, leptin, glucagon like peptide-1, growth hormone, prolactin) through kinase-dependent signalling pathways (PI3-K, p38MAPK, PKA, CaMK) modulating their affinities for DNA and/or for each other. From the overview of the research presented, it is clear that much more study is required to fully comprehend the mechanisms involved in the regulated-expression of the insulin gene in the beta cell to prevent its impairment in diabetes. PMID- 11914737 TI - Concentrations of proinsulin like molecules predict coronary heart disease risk independently of insulin: prospective data from the Caerphilly Study. AB - AIMS/HYPOTHESIS: Higher concentrations of insulin correlate with several coronary heart disease (CHD) risk factors and have been shown to predict incident CHD in several studies, leading to hypotheses concerning the proatherogenic properties of insulin. However, in cross-sectional studies, relationships of concentrations of the insulin precursor molecules, proinsulin and des 31, 32 proinsulin, relate as strongly, or more strongly, to levels of risk factors and to (prevalent) CHD. METHODS: We investigated the relationship between concentrations of insulin, measured with a specific assay, and of proinsulin-like molecules, and risk factors in 1181 non-diabetic men 50-64 years old during Phase II of the Caerphilly Study. We also related concentrations of these molecules to incident CHD during the 10-14 years follow-up. RESULTS: The relationship between concentrations of insulin, of proinsulin and of des 31, 32 proinsulin and BMI ( r = 0.36-0.45), triglyceride (r = 0.25-0.31), high density lipoprotein- (HDL-) cholesterol (r = -0.17 to -0.21), systolic ( r = 0.05-0.11) and diastolic blood pressure ( r = 0.11-0.15) were similarly close, those with risk factors being somewhat and similarly reduced after adjustment for BMI. The correlation between insulin and of proinsulin-like molecules and those plasminogen activator inhibitor-1 (PAI-1) antigen was also similar (r = 0.28-0.29). There was a negative correlation between concentrations of proinsulin-like molecules - but not insulin - and birth weight. Insulin concentrations correlated positively with height ( r = 0.12). In logistic regression models, concentrations of proinsulin like molecules, but not insulin, predicted incident of CHD over a follow-up of 10 14 years (insulin - standardised odds ratio (SOR) 1.30 (95 %-CI) 0.91, 1.85), p = 0.15; des 31, 32 proinsulin - SOR 1.38 (95 %-CI 1.02, 1.85), p = 0.034; sum of proinsulin-like molecules - SOR 1.54 (95 %-CI 1.07, 2.20), p = 0.019 after adjusting for age and BMI. The predictive ability of these molecules was reduced by around one third after adjustment for standard risk factors and concentrations of tryglyceride and HDL-cholesterol, and by about half after further adjustment for PAI-1 concentrations. CONCLUSION/INTERPRETATION: We conclude that concentrations of proinsulin-like molecules provide a better way to predict the incidence of CHD than those of insulin. However, the lack of biological evidence for a causative relationship suggests an association through a common antecedent, and this antecedent is not likely to be intrauterine growth retardation. PMID- 11914738 TI - Increased end-stage diabetic nephropathy in Indo-Asian immigrants living in the Netherlands. AB - AIMS/HYPOTHESIS: We aimed to investigate the risk of end-stage diabetic nephropathy due to Type II (non-insulin-dependent) diabetes mellitus in Indo Asian immigrants from Surinam. METHODS: A demographically based case-control study was carried out in Surinamese Indo-Asian immigrants and Dutch Caucasian subjects. All patients with end-stage diabetic nephropathy who had started dialysis between 1990 and 1998 were identified through a national registry of all patients entering a renal replacement program in the Netherlands. The general population of native Dutch and Surinamese Indo-Asians were considered the control subjects. RESULTS: Among Indo-Asian immigrants, the age adjusted relative risk of end-stage diabetic nephropathy was 38 (95 % CI 16 to 91) compared with the native Dutch population. The duration of diabetes until the start of dialysis treatment was similar in both ethnic groups, about 17 years. CONCLUSION/INTERPRETATION: The Indo-Asian subjects had a nearly 40-fold increase in the risk for end-stage diabetic nephropathy due to Type II diabetes, compared with the native Dutch population. This was higher than expected on the basis of the eightfold higher prevalence of diabetes in the Indo-Asian population. The similar duration of diabetes until the start of dialysis treatment in both ethnic groups supports the hypothesis of a higher incidence of diabetic nephropathy in the Indo-Asian diabetic population. Early and frequent screening for diabetes and microalbuminuria is recommended in Indo-Asian subjects. PMID- 11914739 TI - Effects of size at birth and childhood growth on the insulin resistance syndrome in elderly individuals. AB - AIMS/HYPOTHESIS: A study of 7086 men and women born in Helsinki, Finland, has shown that the development of Type II (non-insulin-dependent) diabetes mellitus is associated with low birth weight followed by accelerated gain in height and weight during childhood and with high maternal BMI but the processes which underlie these associations are largely not known. METHODS: We carried out standard oral glucose tolerance tests, and measured plasma insulin and proinsulin, serum lipid concentrations and blood pressure in 474 patients from the Helsinki cohort. RESULTS: We used four indices of insulin resistance: fasting and 2-h plasma insulin, and fasting proinsulin and 32-33 split proinsulin concentrations. These were associated with small body size at birth and during childhood, rapid growth in height and low maternal BMI. CONCLUSION/INTERPRETATION: Insulin resistance and Type II diabetes share common associations with retarded fetal growth and accelerated growth during childhood. They are dissimilar, however, in that insulin resistance is associated with thinness in childhood and low maternal BMI, while Type II diabetes is associated with high BMI in childhood and high maternal BMI. PMID- 11914740 TI - Components of the insulin resistance syndrome in seven-year-old children: relations with birth weight and the polyunsaturated fatty acid content of umbilical cord plasma phospholipids. AB - AIMS/HYPOTHESIS: According to the foetal origins hypothesis, there is a relation between foetal nutrition and adult glucose intolerance. In adults, insulin resistance has been associated with dietary polyunsaturated fatty acids. We examined whether the availability of polyunsaturated fatty acids during foetal life, as indicated by the fatty acid composition of cord blood samples, relates to childhood body composition and glycaemic control. METHODS: Fatty acid concentrations in umbilical cord blood were determined by gas-liquid chromatography in a birth-cohort of infants. When the children were seven years old, fasting glucose, insulin, proinsulin, and leptin levels were measured in 259 of these children, and relations with cord plasma fatty acid concentrations were studied. RESULTS: Cord plasma phospholipid gamma-linolenic acid and dihomo- gamma linolenic acid concentrations were negatively related to insulin concentrations and calculated insulin resistance (homeostasis model assessment) at seven years of age. The gamma-linolenic acid concentrations were also negatively related to body fatness and proinsulin and leptin concentrations at seven years of age. No association was found for other polyunsaturated fatty acid concentrations at birth. Adjusted for age, sex, current weight, and gestational age, a lower birth weight related to higher values of insulin resistance variables. The highest insulin concentrations were found in children with a low birth weight and a low gamma-linolenic acid concentration at birth. The relations between gamma linolenic acid concentration at birth and fasting insulin and calculated insulin resistance remained statistically significant when adjusted for birth weight. CONCLUSION/INTERPRETATION: These findings indicate that foetal availability or metabolism of gamma-linolenic acid could be involved in the early origins of insulin resistance. PMID- 11914741 TI - Prolonged glucose infusion into conscious rats inhibits early steps in insulin signalling and induces translocation of GLUT4 and protein kinase C in skeletal muscle. AB - AIMS/HYPOTHESIS: Previous studies on diabetic patients have shown that hyperglycaemia increases glucose uptake in an apparently insulin-independent manner. However, the molecular mechanism has not been clarified. METHODS: We studied rats receiving continuous glucose infusion to address this question. In this animal model, rats accommodate systemic glucose oversupply and rapidly develop insulin resistance. RESULTS: Glucose infusion increased both plasma glucose and insulin concentrations to peak after one day. In spite of continuous glucose infusion normoglycaemia was reached after 5 days while insulin concentrations remained higher. Focusing our studies in day 2 (hyperglycaemia/hyperinsulinaemia) and day 5 (normoglycaemia/hyperinsulinaemia) we found, particularly in day 5, that the early steps of the insulin signalling cascade in skeletal muscle of glucose-infused rats were not more activated when compared to control animals as assessed by a comparable phosphorylation of the insulin receptor, IRS-1 and PKB and by an unaltered IRS-1-associated Ptd(Ins) 3' kinase activity. Continuous glucose infusion induced GLUT4 protein expression and translocation to the plasma membrane while neither expression nor translocation of GLUT1 was affected. Translocation of PKC- betaI, - betaII (> threefold) and alpha, -theta (to a lesser extent) to the plasma membrane was significantly induced after 2 days but not after 5 days of glucose infusion when normoglycaemia was reached. CONCLUSIONS/INTERPRETATION: Our data support the hypothesis that continuous glucose infusion induces translocation of GLUT4 while the early steps of the insulin signalling cascade were not increased. These effects could be mediated by activation of PKC. PMID- 11914742 TI - Substituting dietary saturated fat with polyunsaturated fat changes abdominal fat distribution and improves insulin sensitivity. AB - AIMS/HYPOTHESIS: British dietary recommendations are to decrease total fat intake to less than 30 % of daily energy intake and saturated fat to less than 10 %. In practice, it is difficult for people to make these changes. It may be easier to encourage people to switch from a diet rich in saturated fatty acids to one rich in polyunsaturated fatty acids. METHODS: A total of 17 subjects - six people with Type II (non-insulin-dependent) diabetes mellitus, six non-obese and five obese people without diabetes - were randomised to spend two 5-week periods on a diet rich in saturated or in polyunsaturated fatty acids, in a crossover design. At the start of the study and after each dietary period, we assessed abdominal fat distribution using magnetic resonance imaging, insulin sensitivity using hyperinsulinaemic-euglycaemic clamps and fasting lipid parameters. RESULTS: Dietary compliance, assessed by weekly 3-day dietary records and measurement of biochemical markers, was good. Energy and fat intake appeared to be reduced on the diet rich in polyunsaturated fatty acids although body weights did not change. Insulin sensitivity and plasma low density lipoprotein cholesterol concentrations improved with the diet rich in polyunsaturated fatty acids compared with the diet rich in saturated fatty acids. There was also a decrease in abdominal subcutaneous fat area. CONCLUSION/INTERPRETATION: If this result is confirmed in longer-term studies, this dietary manipulation would be more readily achieved by the general population than the current recommendations and could result in considerable improvement in insulin sensitivity, reducing the risk of developing Type II diabetes. PMID- 11914743 TI - Insulin sensitivity is impaired in heterozygous carriers of lipoprotein lipase deficiency. AB - AIMS/HYPOTHESIS: Several studies have investigated the lipoprotein phenotype in heterozygous carriers of a defective lipoprotein lipase allele. We studied whether heterozygosity for lipoprotein lipase deficiency also affects glucose metabolism beyond its effect on plasma lipids. METHODS: To address this question 85 heterozygous carriers of either a missense mutation (Gly188Glu) or a splice site mutation (C-->A in position -3 at the acceptor splice site of intron 6) in the LPL gene which both result in a catalytically inactive product were compared with 108 unaffected subjects from the same families. RESULTS: Carriers for one of these mutations had higher fasting insulin levels but only a trend towards increased fasting blood glucose concentrations could be detected. HOMA index values were significantly higher in carriers than in non-carriers. Furthermore, in carriers, a significantly higher BMI and a trend towards higher systolic and diastolic blood pressure were observed. Carriers also had significantly higher fasting triglycerides, lower HDL cholesterol, and lipoprotein lipase particles of smaller size, confirming previous reports. Among carriers, subjects with one rare allele of the SstI polymorphism in the apo CIII gene had significantly higher plasma triglyceride levels than those with two common SstI alleles. This difference could not be observed in non-carriers of a mutant lipoprotein-lipase allele. The mean intima media thickness of the carotid arteries was slightly, but not significantly higher in carriers when compared with non-carriers. CONCLUSION/INTERPRETATION: This study shows that carrier status of one defective lipoprotein-lipase allele is associated with impaired insulin sensitivity, an atherogenic lipoprotein profile and other characteristics of the metabolic syndrome, which are risk factors for atherosclerotic vascular disease. A higher incidence of atherosclerotic vascular disease, however, could not be firmly established in carriers of this study population. PMID- 11914744 TI - IFN gamma/TNF alpha synergism in MHC class II induction: effect of nicotinamide on MHC class II expression but not on islet-cell apoptosis. AB - AIMS/HYPOTHESIS: Interferon-gamma (IFN gamma) and TNFalpha synergistically induce pancreatic beta-cell apoptosis. Apart from their direct effect, we studied the possible indirect immunological role of IFNgamma/TNFalpha synergism on pancreatic beta-cell death by investigating MHC class II induction by cytokines. The effect of nicotinamide on the cytokine-induced MHC class II expression and pancreatic beta-cell death was also studied. METHODS: Immunocytochemistry, flow cytometry and RNase protection assay were used to study MHC class II expression. Immunoblotting was done to study downstream signals of IFN gamma. The effects of nicotinamide on islet-cell apoptosis and diabetes mellitus were examined using MTT assay and adoptive transfer model. RESULTS: IFN gamma alone induced MHC class II expression on a small number of insulinoma cells. TNFalpha alone did not induce MHC class II expression, but enhanced IFN gamma-induced MHC class II expression. MHC class II expression by cytokine(s) was due to the induction of class II transactivator (CIITA). Nicotinamide reduced MHC class II expression by cytokine(s) but did not protect insulinoma-cell apoptosis by IFN gamma and TNFalpha in combination or protect against the development of diabetes mellitus after adoptive transfer of diabetogenic lymphocytes. CONCLUSION/INTERPRETATION: IFN gamma and TNFalpha synergistically induced MHC class II expression on insulinoma cells through the induction of CIITA; nicotinamide reduced the expression of cytokine-induced MHC class II expression on insulinoma cells through its effect on CIITA expression; and the preventive effect of nicotimamide on Type I (insulin-dependent) diabetes mellitus is probably due to its effect of MHC class II expression rather than that on islet cell apoptosis. PMID- 11914745 TI - Endocrine pancreas development is altered in foetuses from rats previously showing intra-uterine growth retardation in response to malnutrition. AB - AIMS/HYPOTHESIS: We have shown that perinatal malnutrition decreases beta-cell mass at birth and impairs the adaptation of the endocrine pancreas to a subsequent pregnancy. The aim of this study is to investigate the impact of this maternal inadaptation on the development of endocrine pancreas in foetuses. METHODS: Female rats malnourished during their perinatal life and showing intra uterine growth retardation at birth were mated at 8 months of age. The development of the endocrine pancreas was studied at embryonic days 14, 17 and 20 in their foetuses by immunohistochemistry and morphometrical measurements on pancreatic sections. RESULTS: At embryonic day 20, both alpha and beta-cell fractions were decreased in foetuses from IUGR dams. Beta-cell mass was reduced (197 +/- 27 microg, vs 281 +/- 40 microg in control, p < 0.01) and so were insulin content and islet number per cm(2), as in the first generation foetuses. At embryonic day 14, the number of cells expressing only insulin was decreased by half in foetuses from intra-uterine growth retardation dams. At embryonic day 17, 50 % of the homeodomain protein Pdx-1 cell population expressed insulin but all the insulin cells expressed Pdx-1 in both groups; in foetuses from intra-uterine growth retardation dams the number of epithelial cells expressing Pdx-1 was decreased (415 +/- 40 cells/ mm(2) vs 481 +/- 28 cells/mm(2) in control foetuses, p < 0.05) and the mesenchymal fraction in the pancreas was increased by 36 % ( p < 0.05). CONCLUSION/INTERPRETATION: Early malnutrition decreases beta-cell mass in the first generation of offspring and impairs the subsequent beta-cell adaptation to pregnancy. The beta-cell alteration is also present in the next generation and involves a decreased expansion of the epithelial population expressing Pdx-1. PMID- 11914746 TI - Long-acting calcium channel blocker benidipine suppresses expression of angiogenic growth factors and prevents cardiac remodelling in a Type II diabetic rat model. AB - AIMS/HYPOTHESIS: Calcium channel blockers, widely used for the treatment of hypertension and angina, could prevent cardiovascular complications in patients with diabetes. They can improve cardiac remodelling in animal models of a variety of cardiovascular diseases. Here, we examined the therapeutic effect of benidipine, a long-acting calcium channel blocker, on cardiac remodelling in Otsuka-Long-Evans-Tokushima-Fatty (OLETF) rats, a Type II (non-insulin-dependent) diabetes mellitus model. METHODS: The methods for morphometric analysis included double staining for coronary capillaries, dye-binding staining for collagen content and Masson's trichrome staining for perivascular fibrosis. Immunohistochemical and in situ hybridization techniques were used for detecting protein and mRNA expressions for vascular endothelial growth factors (VEGF), basic fibroblast growth factors (bFGF) and TGF-beta(1), endothelial nitric oxide synthase (eNOS), and anti- and pro-apoptotic markers. RESULTS: OLETF rats showed an increased coronary capillary density, a reduced venular capillary proportion, an increased cardiac collagen content and prominent cardiac perivascular fibrosis. In OLETF rat hearts, significant increases in vascular expressions for VEGF, bFGF and TGF- beta(1) were found. Furthermore, the apoptosis signalling pathways, involving eNOS and apoptotic markers, were markedly altered, and coronary endothelial cell apoptosis was lower. These alterations with the exception of eNOS expression were significantly blocked by benidipine treatment. CONCLUSION/INTERPRETATION: The suppressive effect of benidipine on overproduction of angiogenic growth factors could prevent cardiac angiogenesis and fibrosis, resulting in an improvement of cardiac remodelling in diabetes. As VEGF and bFGF potently block endothelial cell apoptosis execution, physiological apoptosis revived by benidipine treatment could also contribute to coronary vessel regression. PMID- 11914747 TI - Pericyte adhesion is impaired on extracellular matrix produced by endothelial cells in high hexose concentrations. AB - AIMS/HYPOTHESIS: Thickening of the basement membrane and selective loss of pericytes are early events in diabetic retinopathy. We aimed at checking whether pericyte interaction with extracellular matrix produced by endothelial cells is influenced by the hexose concentrations in which endothelial cells are cultured. METHODS: Conditioned extracellular matrixes were obtained by growing human umbilical vein endothelial cells in media containing 28 mmol/l hexoses (D glucose, D-galactose, L-glucose), which undergo different intracellular processing, before and after adding the inhibitors of protein glycation thiamine or aminoguanidine. Having removed the endothelium, bovine retinal pericytes were grown on such matrixes and, in separate experiments, on laminin, fibronectin or type IV collagen. Pericyte adhesion was determined by cell counts 18 h after seeding. RESULTS: Reduced adhesion was observed on matrixes produced in high D glucose, high D-galactose and high L-glucose. Both thiamine and aminoguanidine restored impaired pericyte adhesion when added to high D-glucose and high D galactose, but not L-glucose. Laminin, fibronectin and type IV collagen did not consistently modify pericyte adhesion. CONCLUSIONS/INTERPRETATIONS: Pericyte adhesion is impaired on extracellular matrix produced by endothelium in high hexose concentrations. This could result from excess protein glycation, corrected by aminoguanidine and thiamine, rather than altered glycoprotein composition. PMID- 11914748 TI - Coenzyme Q(10) improves endothelial dysfunction of the brachial artery in Type II diabetes mellitus. AB - AIM/HYPOTHESIS: We assessed whether dietary supplementation with coenzyme Q(10) improves endothelial function of the brachial artery in patients with Type II (non-insulin-dependent) diabetes mellitus and dyslipidaemia. METHODS: A total of 40 patients with Type II diabetes and dyslipidaemia were randomized to receive 200 mg of coenzyme Q(10) or placebo orally for 12 weeks. Endothelium-dependent and independent function of the brachial artery was measured as flow-mediated dilatation and glyceryl-trinitrate-mediated dilatation, respectively. A computerized system was used to quantitate vessel diameter changes before and after intervention. Arterial function was compared with 18 non-diabetic subjects. Oxidative stress was assessed by measuring plasma F(2)-isoprostane concentrations, and plasma antioxidant status by oxygen radical absorbance capacity. RESULTS: The diabetic patients had impaired flow-mediated dilation [3.8 % (SEM 0.5) vs 6.4 % (SEM 1.0), p = 0.016], but preserved glyceryl-trinitrate mediated dilation, of the brachial artery compared with non-diabetic subjects. Flow-mediated dilation of the brachial artery increased by 1.6 % (SEM 0.3) with coenzyme Q(10) and decreased by -0.4 % (SEM 0.5) with placebo (p = 0.005); there were no group differences in the changes in pre-stimulatory arterial diameter, post-ischaemic hyperaemia or glyceryl-trinitrate-mediated dilation response. Coenzyme Q(10) treatment resulted in a threefold increase in plasma coenzyme Q(10) (p < 0.001) but did not alter plasma F(2)-isoprostanes, oxygen radical absorbance capacity, lipid concentrations, glycaemic control or blood pressure. CONCLUSION/INTERPRETATION: Coenzyme Q(10) supplementation improves endothelial function of conduit arteries of the peripheral circulation in dyslipidaemic patients with Type II diabetes. The mechanism could involve increased endothelial release and/or activity of nitric oxide due to improvement in vascular oxidative stress, an effect that might not be reflected by changes in plasma F(2) isoprostane concentrations. PMID- 11914750 TI - Taq I polymorphism of the vitamin D receptor and risk of severe diabetic retinopathy. AB - AIMS/HYPOTHESIS: Vitamin D, a molecule with antiproliferative, antiangiogenic, antioxidant and immunosuppressive effects, could play a role in the pathogenesis of severe diabetic retinopathy. We examined whether Taq I polymorphism of the vitamin D receptor is involved in the development of severe diabetic retinopathy. METHODS: 200 unrelated C-peptide-negative French Type I diabetic patients were randomly selected (male:female, 103:97, age 44.4 +/- 12.4 years, diabetes duration: 27.7 +/- 10.0 years, BMI: 24.3 +/- 3.4 kg/m(2), HbA(1c): 8.6 +/- 1.3 %). The Taq I site was analysed by PCR followed by digestion with Taq I enzyme. Diabetic retinopathy was assessed by retinal angiography and classified as presence (n = 101) or absence (n = 99) of severe (preproliferative or proliferative) diabetic retinopathy. RESULTS: Frequency of wild-type genotype TT was lower in patients with severe diabetic retinopathy (n = 27) when compared with control subjects (n = 42, OR = 0.5, p = 0.028). Allele frequencies were not different between patients (T: n = 112 and t: n = 90) and control subjects (T: n = 128, and t: n = 70, p = 0.075). Global chi(2) (df = 2): p = 0.064. In subjects with diabetes duration of more than 25 years, TT was lower in severe diabetic retinopathy (n = 14) than control subjects (n = 18, OR = 0.3, p = 0.01). Allele frequencies were different between patients (T: n = 68 and t: n = 66) and control subjects (T: n = 52, OR = 0.5, and t: n = 26, OR = 1.9, p = 0.034). Global chi(2) (df = 2): p = 0.024. In subjects with HbA(1c) over 9 %, Tt was higher in patients (n = 28) than control subjects (n = 15, OR = 3.1, p = 0.019). Allele frequencies were not different between patients (T: n = 52 and t: n = 38) and control subjects (T: n = 57, and t: n = 29, p = 0.31). Global chi(2) (df = 2): p = 0.035. CONCLUSION/INTERPRETATION: In French Type I (insulin-dependent) diabetic patients, we demonstrate an association between TT form (VDR) and low risk for severe diabetic retinopathy, especially in patients with long duration, and between Tt variant and high risk for severe diabetic retinopathy in subjects with poor glycaemic control. PMID- 11914749 TI - The genetic abnormality in the beta cell determines the response to an oral glucose load. AB - AIMS/HYPOTHESIS: We assessed how the role of genes genetic causation in causing maturity-onset diabetes of the young (MODY) alters the response to an oral glucose tolerance test (OGTT). METHODS: We studied OGTT in 362 MODY subjects, from seven European centres; 245 had glucokinase gene mutations and 117 had Hepatocyte Nuclear Factor -1 alpha ( HNF-1alpha) gene mutations. RESULTS: BMI and age were similar in the genetically defined groups. Fasting plasma glucose (FPG) was less than 5.5 mmol/l in 2 % glucokinase subjects and 46 % HNF-1 alpha subjects ( p < 0.0001). Glucokinase subjects had a higher FPG than HNF-1 alpha subjects ([means +/- SD] 6.8 +/- 0.8 vs 6.0 +/- 1.9 mmol/l, p < 0.0001), a lower 2-h value (8.9 +/- 2.3 vs 11.2 +/- 5.2 mmol/l, p < 0.0001) and a lower OGTT increment (2-h - fasting) (2.1 +/- 2.3 vs 5.2 +/- 3.9 mmol/l, p < 0.0001). The relative proportions classified as diabetic depended on whether fasting (38 % vs 22 %, glucokinase vs HNF-1 alpha) or 2-h values (19 % vs 44 %) were used. Fasting and 2-h glucose values were not correlated in the glucokinase subjects ( r = 0.047, p = 0.65) but were strongly correlated in HNF-1 alpha subjects ( r = 0.8, p < 0.001). Insulin concentrations were higher in the glucokinase subjects throughout the OGTT. CONCLUSION/INTERPRETATION: The genetic cause of the beta cell defect results in clear differences in both the fasting glucose and the response to an oral glucose load and this can help diagnostic genetic testing in MODY. OGTT results reflect not only the degree of hyperglycaemia but also the underlying cause. PMID- 11914752 TI - Self-assembled "nanocubicle" as a carrier for peroral insulin delivery. AB - AIMS/HYPOTHESIS: A patient with (insulin-dependent) diabetes mellitus receives at least one subcutaneous insulin injection a day to maintain low serum glucose concentrations. Since patients' compliance with such dosage regimens is too low, the development of an oral formula is clearly attractive. We present the development of a liquid formula that can be easily dispersed in water to produce particles named "nanocubicles" which efficiently encapsulate insulin. METHODS: Fasted streptozotocin-induced diabetic rats were administered orally with particles encapsulating insulin, and particles without insulin or soluble insulin in water. Groups of rats were also injected soluble insulin in PBS for control. Blood glucose concentration and insulin concentration were measured 1, 2, 3, 4 and 6 h after the administration of the insulin formulas. RESULTS: In vitro experiments show that the particles can be taken up by the Caco-2 cells at a high ratio. The serum glucose concentration was controlled for more than 6 h after oral insulin administration but returned to the basal concentration in 3 h when 1 IU/kg of insulin was injected intravenously. CONCLUSION/INTERPRETATION: Our biocompatible and stable oral insulin formulation is easy to prepare and produces reproducible hypoglycaemic effects, therefore we anticipate clinical acceptance and utilization of this form of insulin therapy. PMID- 11914751 TI - A retroviral long terminal repeat adjacent to the HLA DQB1 gene (DQ-LTR13) modifies Type I diabetes susceptibility on high risk DQ haplotypes. AB - AIMS/HYPOTHESIS: HLA-DQ genes, located in the human leukocyte antigen region on chromosome 6 p, are the main inherited factors predisposing to Type I (insulin dependent) diabetes mellitus. Endogenous retroviral long-terminal repeats are integrated at several sites within this region, one of which is known to enhance susceptibility for Type I diabetes. We examined another LTR within the HLA-region as an additional genetic risk marker. METHODS: We investigated the segregation of one long-terminal repeat (DQ-LTR13), located 1.3 kb upstream of HLA DQB1 with different HLA-DQ haplotypes, and its transmission to patients. A total of 284 Caucasian families (203 German and 81 Belgian) with at least one diabetic offspring were genotyped for DQA1, DQB1 and DQ-LTR13. RESULTS: DQ8/LTR13(+) was preferentially transmitted (139 transmitted vs 28 not transmitted; P(TDT) = 1.67 x 10(-14)) whereas no deviation from expected transmission frequencies was observed for DQ8/LTR13(-) (20 transmitted vs 17 not transmitted; P(TDT) = 1.00). DQ8/LTR13(+) alleles conferred a significantly higher risk for Type I diabetes than DQ8/LTR13(-) alleles (p chi(2) = 2.58 x 10(-14)). This difference remained significant even after DRB1 subtyping (p chi(2) = 0.02). Also, there was a significant difference when comparing the transmission of DQ2/LTR13(+) and DQ2/LTR13(-) alleles (p chi(2) = 0.01), the latter conferring an increased risk. The transmission of DQ-LTR13(+) haplotypes did not show any differences regarding paternal, maternal or gender-related stratification. However, DQ8/LTR13(-) was significantly more often transmitted from mothers (p chi(2) = 0.01) and to female patients (p chi(2) = 0.04). CONCLUSION/INTERPRETATION: We conclude that DQ-LTR13 marks additional genetic risk for Type I diabetes on predisposing DRB1(*)0401- DQ8 and DQ2 haplotypes and will help to further define susceptibility in this gene region. PMID- 11914753 TI - HLA class II alleles and haplotypes in Icelandic Type I diabetic patients: comparison of Icelandic and Norwegian populations. PMID- 11914754 TI - IL-6 gene polymorphism -174G/C does not contribute substantially to hyperlipidaemia and Type II diabetes mellitus in Japanese men. PMID- 11914755 TI - Glucokinase gene mutations are not a common cause of permanent neonatal diabetes in France. PMID- 11914756 TI - Detecting glucose intolerance after gestational diabetes: inadequacy of fasting glucose alone and risk associated with gestational diabetes and second trimester waist-hip ratio. PMID- 11914757 TI - Incidence of childhood Type I diabetes mellitus is increasing at all ages in Germany. PMID- 11914760 TI - Report of the 8th EASD-DOIT Study Group Workshop "Decision Support in Diabetes Care" Stirling Royal Infirmary, Stirling, UK, 7-8 September 2001. PMID- 11914761 TI - Thermal shrinkage of the anterior cruciate ligament: any good? PMID- 11914762 TI - Tightening elongated ACL grafts by application of bipolar electromagnetic energy (ligament shrinkage). AB - In ACL transplants showing elongation with functional deficit but no interruption of structure (Marburg Arthroscopic Score type III) there is the possibility of reinsertion, augmentation, or revision ACL reconstruction to stabilize the joint. Bipolar electromagnetic energy can be used to induce the tightening of elongated ACL transplants. We examined 14 patients with a secondary instability after ACL transplantation (mean 23.6 months). During arthroscopy a type III transplant without signs of nonisometric position was found. The elongated transplants were treated by bipolar application probe. All knees where protected by an unlimited orthosis over a period of 12 weeks. Postoperative follow-up took 9.4 months. There was a significant improvement in subjective discomforts (35.9 to 88.5 points on the Lysholm score), and the Rollimeter test showed a reduction of the ventral tibial translation (13.1 to 2.6 mm). There was a correlation between the degree of chondral damage and a Lysholm score but not between subjective discomforts and the degree of instability. Our findings confirm that elongated ACL transplants can be tightened by applying bipolar electromagnetic energy during rearthroscopy, thus avoiding revision ACL reconstruction in some cases. PMID- 11914763 TI - Patellar tendon anterior cruciate ligament reconstruction with conical press-fit femoral fixation: 5-year results in athletes population. AB - Bone-patellar tendon autograft is probably the most widely used graft for ACL reconstruction. Several methods for graft fixation have been described. To avoid intra-articular hardware we adopt biological fixation with a femoral conical press-fit fixation. A prospective study was performed on 40 consecutive active athletes who underwent ACL reconstruction with this technique by the same surgeon between November 1994 and September 1995 (mean follow-up 46 months, range 36-62). Results were evaluated by an independent examiner using radiography, computed tomography, subjective and objective evaluation, and isokinetic and functional strength tests. Assessment using the IKDC knee scoring revealed 85% of the patients with a normal or nearly normal knee joint; Tegner's score was 7.5 preoperatively and 6.0 postoperatively, with 60% of the athletes returning to the preinjury sport and level. No patients had instability, with 90% having less than 3 mm side-to-side difference on computerized analysis. The isokinetic test showed mild quadriceps deficit at 3 and 6 months, with no deficit at final follow-up; four patients complained of anterior knee pain and had a positive kneeling test. We found no graft dislocation. All cases showed radiological evidence of graft integration at 3 months time. Long-term results support this technique asx a simple, cost-effective, and reliable alternative for patellar tendon fixation in ACL reconstruction. PMID- 11914764 TI - A prospective evaluation of tunnel enlargement in anterior cruciate ligament reconstruction with hamstrings: extracortical versus anatomical fixation. AB - Changes in the femoral and tibial bone tunnel were studied prospectively after arthroscopic ACL reconstruction with quadruple hamstring autograft. To determine whether tunnel enlargement can be decreased by fixing the graft close to the joint line having a stiffer fixation construct we compared "anatomical" (one absorbable interference screw femorally, and bicortical fixation with two absorbable interference screws tibially) and extracortical fixation techniques (Endobutton femorally, and two no. 6 Ethibond sutures over a suture washer tibially). Over a 2-year period we evaluated 60 patients clinically (IKDC scale, Cincinnati Knee Score, KT-1000) and radiographically (confirmed by MRI). The operated knee was radiographed immediately postoperatively and 6 and 24 months postoperatively. The femoral and tibial bone tunnel diameter was measured on anteroposterior and lateral images, and the tunnel area was calculated and compared to the initial area calculated from the perioperative drill size. In the "anatomical" group the immediately postoperative bone tunnel area was 75% larger than the initial tunnel area, after 6 months it was increased another 31%, and between 6 and 24 months it remained basically unchanged. In the "extracortical" group there was no significant enlargement immediately postoperatively, but after 6 months it was 65% larger than the initial area of drill and graft size, and between 6 and 24 months it decreased to 47%. There was no correlation between the amount of tunnel enlargement and clinical scores or KT-1000 measurement. Arthroscopic ACL reconstruction with quadruple hamstring autograft is associated with bone tunnel enlargement. Using a purely extracortical fixation technique thus significantly increased the tibial and femoral tunnel area during the first 6 postoperative months, while it decreased slightly thereafter. The insertion of large interference screws apparently not only compresses the graft in the bone tunnel but also significantly enlarges the bone tunnel itself. The immediate enlargement at the time of the operation is followed by a reduced further enlargement at 6 months and then stabilization. Tunnel widening did not influence clinical outcome over a 2-year period. PMID- 11914765 TI - Posterolateral aspect and stability of the knee joint. I. Anatomy and function of the popliteus muscle-tendon unit: an anatomical and biomechanical study. AB - This study examined ten human adult cadaveric knees to dissect the popliteus muscle-tendon unit (PMTU), including its numerous attachments to other posterior and posterolateral structures, and to determine the effect of tensioning the PMTU on the internal and external rotation, total rotational arcs, and neutral tibial rotation in full extension and 30 degrees, 60 degrees, and 90 degrees of knee flexion. The junction between the popliteus tendon and the fibular head commonly described as the popliteofibular ligament became lax in internal and tense in external tibial rotation. The internal and external rotational arcs increased gradually between extension and 90 degrees of flexion. Tensioning the PMTU resulted in a statistically highly significant internal tibial rotation with decreased internal and increased rotational arcs. The anatomical findings and functional data indicate that the PMTU is an important structure maintaining dorsolateral stability, stabilizing the lateral meniscus, and balancing the neutral tibial rotation. PMID- 11914766 TI - Posterolateral aspect and stability of the knee joint. II. Posterolateral instability and effect of isolated and combined posterolateral reconstruction on knee stability: a biomechanical study. AB - This study evaluated the correlation between the number of transected posterolateral structures (PLS) and the grade of posterolateral rotational instability, determined the effect of the popliteus muscle-tendon unit on the tibial rotation, and examined the effect of an isolated posterior cruciate ligament (PCL) and combined PCL-PLS reconstruction on knee stability. Sectioning the popliteofibular and lateral collateral ligaments both caused an increase in tibial external rotation. Cutting the PT resulted in a statistically highly significant excessive external rotation and externally shifted neutral position of the tibia over the full range of motion. Tensioning the popliteus muscle tendon unit led to a statistically highly significant internally shifted neutral tibial rotation and a decreased internal and an increased external rotation without affecting the total rotational arcs. The isolated PCL reconstruction did not affect the external rotation, whereas the combined PCL-PLS reconstruction reset the knee to nearly physiological laxity patterns. PMID- 11914767 TI - Bone bruise in the acutely injured knee. AB - We used MRI to study the lesions in a consecutive group of 64 patients with an acute trauma of the knee and normal plain radiography during the winter season. Bone bruise was present in 35 of the patients, and these were referred to subsequent MRI 4 and 12 months later. After 4 months bone bruise was still present in 69% of the patients and after 12 months in 12%. Soft tissue lesions, for example, lesion in the meniscus, ACL rupture, or MCL lesion was present in 94% of the patients with bone bruise. Interobserver agreement varied from good to excellent. Soft tissue lesions are found in almost all patients with bone bruise, and these generally resolve 4-12 months after the injury. In the patients with no bone bruise the number of ligament injuries was lower. PMID- 11914768 TI - The tibial attachment of the anterior cruciate ligament in children and adolescents: analysis of magnetic resonance imaging. AB - Recent studies have demonstrated that skeletally immature athletes with an ACL injury may require surgical reconstruction if they return to high-demand sports. This study used MRI to compare the anatomy of the ACL in skeletally immature and adult subjects. Measurements were recorded in the sagittal plane for the anterior posterior dimension of the proximal tibia, and the anterior, center, and posterior limits of the ACL, and the roof inclination angle of the femur. These values were compared to established reference values for adult knee anatomy. In skeletally immature women ( n=7) the ACL anterior limit, center and posterior limit, and roof inclination angle were 28%, 46%, 63%, and 38 degrees, respectively, compared to 28%, 44%, 60%, and 35 degrees in adult women. In skeletally immature men ( n=15) the ACL anterior limit, center, posterior limit, and roof inclination angle were 27%, 43%, 59%, and 40 degrees, respectively, compared to 28%, 44%, 59%, and 37 degrees in adult men. In the younger subjects the overall dimensions of the proximal tibia were smaller than that in adults, but the anatomical landmarks for the ACL were proportional. If ACL reconstruction is performed in skeletally immature subjects, the smaller dimensions of the tibia need to be considered, and the use of anatomical landmarks is an important factor in graft placement PMID- 11914770 TI - Extensive subcutaneous emphysema following arthroscopy: a case report. AB - We report an unusual presentation of delayed subcutaneous emphysema of the thigh following arthroscopy with a fluid medium, which was managed conservatively. This rare presentation must be recognised by all orthopaedic surgeons practising arthroscopic surgery. PMID- 11914771 TI - Anterior labroligamentous periosteal sleeve avulsion lesion at the superior glenoid labrum. AB - An unusual type of anterior labroligamentous periosteal sleeve avulsion (ALPSA) lesion in the superior, instead of the usual inferior, part of the anterior glenoid labrum is presented in a 33-year-old man who sustained a traumatic dislocation of his left shoulder. Magnetic resonance imaging raised the possibility of an ALPSA lesion and later arthroscopy confirmed the diagnosis. Arthroscopic repair of the lesion was performed after the labral avulsion was lateralized. PMID- 11914769 TI - Joint degeneration following meniscal allograft transplantation in a canine model: mechanical properties and semiquantitative histology of articular cartilage. AB - This study examined the hypothesis that meniscal allograft transplantation serves a "chondroprotective" role and prevents the histological and biomechanical changes of the articular cartilage following meniscectomy. Skeletally mature mongrel dogs underwent total medial meniscectomy and received either a fresh meniscal allograft ( n=10) or no further treatment ( n=10). Semiquantitative histology and biomechanical analysis of the femoral articular cartilage was used to assess cartilage pathology 12 weeks following surgery. Histological analysis showed significant changes in cartilage structure that did not differ between the meniscectomy and allograft transplantation groups. Similarly, the tensile modulus of the surface zone cartilage was significantly lower than that in unoperated controls following either meniscectomy or allograft transplantation. A significant correlation was observed between the biomechanical and histological changes, suggesting that degenerative changes in cartilage structure and mechanical function are interrelated. Our findings do not support the hypothesis that meniscal allograft transplantation provides chondroprotection of the femoral condyle and also suggest that it does not lead to increased degenerative changes. PMID- 11914772 TI - Which joint position puts the axillary nerve at lowest risk when performing arthroscopic capsular release in patients with adhesive capsulitis of the shoulder? AB - The success of arthroscopic capsular release of the glenohumeral joint depends on complete incision of the inferior capsule. This study determined the distance between capsule and the axillary nerve in different joint positions. In 14 human shoulder specimens the anterior joint capsule and axillary nerve were dissected, and the anterior joint capsule was incised between the 1 and 5 o'clock positions. The shortest distance between the insertion of the inferior capsule and the axillary nerve was measured at the glenoid and humeral insertions in abduction, adduction, internal, and external rotation. The axillary nerve is surrounded from soft connective tissue and is closer to the humeral than to the glenoidal attachment of the joint capsule. During abduction and external rotation the nerve stays in its position while the glenohumeral capsule tightens, which increases the distance between the two structures. This results in the following distances: to the glenoidal/humeral capsule insertion: in adduction and neutral rotation, 21.2+/-4.2/14.2+/-2.6 mm; in abduction and neutral rotation, 24.0+/-4.9/15.0+/ 5.0 mm; in abduction and internal rotation, 21.1+/-6.6/14.6+/-3.7 mm; and in abduction and external rotation, 24.9+/-3.8/16.4+/-4.4 mm. Thus, when performing arthroscopic capsular release the incision of the glenohumeral joint capsule should be undertaken at the glenoidal insertion in the abducted and externally rotated shoulder. PMID- 11914773 TI - Endoscopy, assisted percutaneous repair of the Achilles tendon ruptures: a cadaveric and clinical study. AB - Endoscopy-assisted percutaneous repair of Achilles tendon ruptures were investigated in a cadaveric and clinical study. Sixteen above-knee fresh amputation specimens in which different types of Achilles tendon ruptures were created were repaired percutaneously with the visualization of the tendon ends by endoscopy. Neither malalignment nor damage to the neurovascular structures was observed. Eleven patients were treated in this way. No reruptures, wound problems, or neurovascular injury were observed. All patients returned to daily activities 10-11 weeks after the repair. This technique seems to overcome certain problems of conservative, surgical, or percutaneous repair of the Achilles tendon ruptures. PMID- 11914775 TI - Subacute oral toxicity study of ethynylestradiol and bisphenol A, based on the draft protocol for the "Enhanced OECD Test Guideline no. 407". AB - We performed a 28-day repeated-dose toxicity study of ethynylestradiol (EE) and bisphenol A (BPA) based on the draft protocol of the "Enhanced OECD Test Guideline no. 407", and assessed the sensitivity of a list of parameters for detecting endocrine-related effects of endocrine disruption. Doses of EE at 0, 10, 50 or 200 microg/kg per day, or BPA at 0, 40, 200 or 1000 mg/kg per day were orally administered to Sprague-Dawley rats. The highest dose of BPA was decreased to 600 mg/kg per day from the second week of administration because a male rat given 1000 mg/kg BPA had died within 1 week with toxic clinical signs. In the assay using EE, the decrease of prostate, seminal vesicle and pituitary weights, increase of the testis weight, atrophic changes of the prostate, seminal vesicle and mammary gland, and degenerative changes in the testes were detected in male rats in the 50 and/or 200 microg/kg groups. In females of the 200 microg/kg group, decrease of the ovary weight, increase of the uterine weight, atrophy of the ovary, hypertrophy or squamous metaplasia of the uterine epithelial cells and mucification in the vagina were observed. Furthermore, diestrous, estrous or the unknown stage was prolonged in the 50 and 200 microg/kg groups of rats. Endocrine mediated effects of EE were not detected in general observations, hematology, serum biochemistry, or hormonal or spermatological examinations. In the assay using BPA, the diestrous stages were prolonged at the highest dose, but changes related to endocrine effects were not detected in other examinations. Thus, among the parameters tested, the weight of endocrine-linked organs and their histopathological assessment and estrous cycle stage allowed the detection of the endocrine-related effect of EE, whereas the estrous cycle stage was only a useful parameter to detect the effect of BPA. PMID- 11914776 TI - Differential induction of CYP1A1 in duodenum, liver and kidney of rats after oral intake of soil containing polycyclic aromatic hydrocarbons. AB - We have analyzed the induction of the cytochrome P450 enzyme CYP1A1 as a biomarker of effect in duodenum, liver and kidney of rats after oral intake of contaminated soil particles. The soil samples originated from industrial sites and were contaminated with polycyclic aromatic hydrocarbons (PAH) to variable extents, ranging from 60 to 4700 mg PAH/kg soil. Soil samples were administered for one week as a mixture with commercial rodent diets. After exposure, microsomes of several organs were prepared and analyzed for CYP1A1, enzymatically and by Western blots. All contaminated soils led to induction of CYP1A1 in duodenal mucosa cells, regardless of their extent of contamination, showing that relevant doses were mobilized in the gastrointestinal tract and adsorbed. Subsequent distribution of non-metabolized compounds is indicated by induction of CYP1A1 in the liver. However, some samples did not lead to a response in the liver, due to their quantitative and qualitative contaminant composition. In accordance with previous results, there is a sigmoidal dose-response relationship between induction of hepatic CYP1A1 levels and the soil contamination with higher condensates of PAH. In contrast, the response in the duodenum appeared to be hyperbolic and correlated well with the amounts of total PAH. Highly contaminated soil, being nearly devoid of higher condensates of PAH, led to pronounced induction in the duodenum but failed to induce CYP1A1 in the liver. Successful passage of contaminants through the intestinal barrier and the liver compartment is shown by increased CYP1A1 expression in the kidney. Compared with enzyme levels induced in the liver, those of the kidney are much lower and amount to only about 1/20 of the liver values for soils with high induction potential. Hence, oral PAH intake leads to differential induction patterns of CYP1A1 in duodenum, liver and kidney of rats. The observations raise questions concerning the role of the primary duodenal PAH metabolism in preventing contaminant dependent hazardous effects, and of the significance of differential CYP1A1 expressions for carcinogenic processes in several tissues. PMID- 11914777 TI - Covalent binding of nitrogen mustards to the cysteine-34 residue in human serum albumin. AB - Covalent binding of various clinically important nitrogen mustards to the cysteine-34 residue of human serum albumin, in vitro and in vivo, is demonstrated. A rapid method for detection of these adducts is presented, based on liquid chromatography-tandem mass spectrometry analysis of the adducted tripeptide Cys*-Pro-Phe after digestion of the protein with Pronase. PMID- 11914778 TI - Does the anaerobic formation of hydroxyl radicals by paraquat monocation radicals and hydrogen peroxide require the presence of transition metals? AB - This in vitro study investigated the formation of hydroxyl radicals (*OH) under anaerobic conditions through the direct reaction between paraquat radicals (PQ(+)*) and hydrogen peroxide (H(2)O(2)) by quantitative UV-VIS and electron spin resonance (ESR) spectroscopy. PQ(+)* was formed by paraquat reduction using either sodium dithionite or the xanthine/xanthine oxidase reaction as electron donors. The anaerobic formation of PQ(+)* was quantified both by measuring light absorption at 605 nm or by ESR techniques respectively, using either the absorption coefficient or ultramarine as a stable spin standard. Detection of *OH took place with aid of the spin trap 5-diethoxyphosphoryl-5-methyl-1-pyrroline- N oxide (DEPMPO). Generation or addition of H(2)O(2) to PQ(+)* eliminates the 35 line ESR signal of PQ(+)* and subsequently generates the 8-line ESR signal of the DEPMPO-OH adduct. The elimination of PQ(+)* as well as the formation of OH-DEPMPO adduct was not influenced by 1.0 mM deferoxamine, indicating that iron or other transition metals are, at least under anoxic conditions, not necessarily involved in the generation of the most aggressive reactive oxygen species *OH. PMID- 11914779 TI - Effect of immobilization, cold and cold-restraint stress on liver monooxygenase activity and lipid peroxidation of influenza virus-infected mice. AB - The present study provides a direct experimental evidence that the combination of influenza A/Aichi/2/68 (H3N2) infection with different models of "oxidative stress", such as immobilization, cold and cold-restraint, is associated with graduated oxidative disturbances in the liver of mice, despite the absence of virus and inflammation in this tissue. It was found that experimental influenza virus infection is accompanied with a significant increase of lipid peroxidation products, a decrease of natural antioxidants (vitamin E, glutathione) and cytochrome P-450, an inhibition of cytochrome c reductase and liver monooxygenases (analgin- N-demethylase and amidopyrine- N-demethylase). Immobilization and cold stress, applied separately or in combination (cold restraint), did not influence significantly any of the analysed parameters compared to those of the control group of non-infected mice. Preliminary exposure of mice to immobilization or cold stress and subsequent inoculation of influenza virus resulted in a significant increase of lipid peroxidation products and a significant decrease of vitamin E and reduced glutathione, compared with levels in control (non-infected) animals. Compared to influenza virus-infected and non stressed animals, the changes in all these parameters were negligible. Immobilization or cold stress, applied in combination with influenza virus infection, partially prevented the suppressive effect of influenza virus on cytochrome P-450 and liver monooxygenases. A tendency towards normalization of these parameters to the control levels was observed. However, after application of cold-restraint plus influenza virus infection, the level of cytochrome P-450 and activity of cytochrome c reductase stayed markedly lower than in infected and non-stressed animals. The activities of liver monooxygenases were slightly increased compared with those of infected and non-stressed animals, but stayed relatively low compared to control (non-infected) mice. Combination of cold restraint and influenza virus infection resulted in a greater synergistic increase of lipid peroxidation products and a greater synergistic decrease of vitamin E and reduced glutathione compared to controls, as well as to influenza virus-infected and non-stressed animals. PMID- 11914780 TI - Differential effects of olfactory toxicants on olfactory regeneration. AB - The aim was to study the long-term response in the olfactory mucosa of NMRI mice after exposure to the olfactory toxicants dichlobenil (a herbicide) or methimazole (an antithyroid drug). Three and six months after exposure to dichlobenil (2x or 1 x 25 mg/kg i.p.), the dorsomedial part of the olfactory region showed a respiratory metaplasia with abundant invaginations and a fibrotic lamina propria. In contrast, 3 months after exposure to a toxic dose of methimazole (2 x 50 mg/kg i.p.), the olfactory neuroepithelium and lamina propria had been restored. To study the regenerative events, we used an antibody derived against growth-associated protein 43 (GAP-43), which stains immature neurons. To study epithelial differentiation and horizontal basal cells (HBCs) we used an antibody derived against some cytokeratins. Two weeks after methimazole treatment, there was a marked increase of GAP-43-stained cells in the whole olfactory region, which correlated with the observed regeneration at that time. Two weeks after dichlobenil treatment, the damaged atypical epithelium in the olfactory region showed a distinct keratin staining of basal and columnar cells whereas GAP-43-stained cells were not found. Despite a transient increase of GAP 43-stained cells in the border zone between damaged and undamaged olfactory mucosa, an expansion of a normal neuroepithelium into the damaged olfactory region was not detected in the dichlobenil-treated mice. An intact lamina propria is suggested as a prerequisite for repopulation of the neuroepithelium after toxicant-induced injury. PMID- 11914782 TI - Topical treatment with povidone iodine reduces nitrogen mustard-induced skin collagenolytic activity. AB - Recently we have shown that post-exposure treatment with povidone iodine (PI) protects against nitrogen and sulfur mustard-induced skin lesions. Since proteolytic activity is involved in skin damage caused by chemical irritants, we have studied the effect of iodine on mechlorethamine (HN2)-induced skin collagenolytic activities in the haired guinea pig model. The matrix metalloproteinase-9 (MMP-9) activity increased by 30, 46, 12 and 23% after 3, 24, 48 and 72 h of HN2 exposure, respectively, whereas the MMP-2 was elevated by 8, 65, 8 and 30%, respectively. Topical treatment with PI at 15 and 120 min after HN2 exposure decreased the MMP-9 activity by 67% and 60%, respectively, when skin was analyzed 3 h after exposure. The same trend was observed in the MMP-2 and MMP 1 activities after PI treatment. A stronger effect of PI treatment 15 min following exposure was observed in skin analyzed 24 h after exposure, i.e. a decrease of 83% and 88% in MMP-9 and MMP-2 activities, respectively. Similar findings were observed with an interval of 120 min between HN2 exposure and PI treatment. A much weaker effect was observed on MMP-1 activity. A similar trend of PI-induced reduction in the three types of collagenase activity was found in skin analyzed 48 and 72 h after exposure. Reduced collagenolytic activity may serve as one of the mechanisms by which iodine protects the skin against chemical insult. PMID- 11914781 TI - Induction of oxidative stress in rat epididymal sperm after exposure to 2,3,7,8 tetrachlorodibenzo-p-dioxin. AB - The ability of 2,3,7,8-tetrachlorodibenzo- p-dioxin (TCDD) to induce oxidative stress in various tissues of animals has been reported. The nature and mechanism of action of TCDD on the antioxidant system of sperm has not been studied. In the present study we have sought to investigate whether TCDD induces oxidative stress in the epididymal sperm of rats. Subchronic doses of TCDD (1, 10, and 100 ng/kg body weight per day) were administered orally to male Wistar strain rats for 45 days. After 24 h of the last treatment the rats were killed using diethyl ether. The epididymides were removed and cleared from the adhering tissues. Epididymal sperm were collected by cutting the epididymides into small pieces in Ham's F12 medium, and counted using a hemocytometer. The epididymal sperm counts in the TCDD-treated groups decreased in a dose-dependent manner from the control value of 8.2+/-0.14 x 10(8) to 5.31+/-0.15 x 10(8). Since a positive correlation (r=0.95; n=24) was observed between sperm count and DNA content of the epididymal sperm, DNA content was routinely used as an indicator of sperm count, and the results were expressed in terms of both protein and DNA. There was a significant decline in the activities of superoxide dismutase (40+/-2.17 to 27.1+/-0.76/mg protein and 32.41 to 18.07+/-0.76/mg DNA), catalase (2.49+/-0.13 to 2.03+/ 0.05/mg protein and 2.01+/-0.05 to 1.35+/-0.05/mg DNA), glutathione reductase (71.2+/-3.87 to 48+/-1.79/mg protein and 57.58+/-1.52 to 31.94/mg DNA) and glutathione peroxidase (22.4+/-1.43 to 16.9+/-1.57/mg protein and 18.08+/-0.61 to 11.38+/-1.22/mg DNA) while there were increases in the levels of hydrogen peroxide (20.8+/-1.96 to 55.3+/-0.88/ mg protein and 16.18+/-1.88 to 36.87+/ 0.88/ mg DNA) and lipid peroxidation (2.17+/-0.2 to 6.08/mg protein and 1.75+/ 0.12 to 4.05+/-0.12/mg DNA) in the epididymal sperm. The results suggest that graded doses of TCDD elicit depletion of antioxidant defense system in sperm, indicating TCDD-induced oxidative stress in the epididymal sperm. In conclusion, the adverse effect on male reproduction in TCDD-treated rats may be due to the induction of oxidative stress in sperm. PMID- 11914783 TI - Genotoxicity study of a new tetraalkylammonium derivative of 6-methyluracil (agent No. 547). AB - Agent No. 547 (1,3-bis[omega-(diethyl-ortho-nitrobenzylammonio)-pentyl]-6 methyluracil dibromide), a newly synthesized inhibitor of mammalian-specific acetyltcholinesterase (EC 3.1.1.7) was investigated for genotoxicity using the DNA-repair test, Ames test and in vivo micronucleus test with mouse peripheral blood erythrocytes. Agent No. 547 did not cause significant changes in growth of repair-deficient Escherichia coli tester strains. The compound was non-mutagenic in Salmonella typhimurium strains TA98 and TA100 with and without rat microsomal activation mixture. However, we observed a marked increase in number of His(+) revertants for both tester strains in preincubation assays. The results obtained in the micronucleus test indicate that agent No. 547 possesses significant clastogenic activity. At the high dose tested (0.5 mg/kg), the compound induced a seven-fold increase in the number of micronuclei over the spontaneous background 48 h after treatment. The results suggest that further work should be promoted to identify the metabolic pathways involved in genotoxicity of agent No. 547 in mammalian cells and to evaluate the real risk of its exposure. PMID- 11914784 TI - Generation of the quick phase of horizontal vestibular nystagmus. AB - This paper reviews evidence concerning the anatomy and physiology of brainstem neurons causing the quick phase of horizontal vestibular nystagmus in the cat. Available anatomical and physiological evidence is used to show for the first time that presently known neural connections constitute a network which results in the generation of the quick phase during maintained horizontal angular accelerations. PMID- 11914785 TI - Adaptation to destabilizing dynamics by means of muscle cocontraction. AB - Adaptive control of wrist mechanics was investigated by means of destabilizing dynamics created by a torque motor. Subjects performed a 20 degrees movement to a 3 degrees target under the constraint that no motion should occur outside of the target zone once 800 ms had elapsed from movement onset. This constraint served as the minimum acceptable level of postural stability. The ability of subjects to modify their muscle activation patterns in order to successfully achieve this stability was investigated by creating three types of destabilizing dynamics with markedly different features: negative stiffness, negative damping, and square wave vibration. Subjects performed sets of trials with the first type of destabilizing dynamics and were then required to adapt to the second and third. The adaptive response was quantified in terms of the rms electromyographic (EMG) activity recorded during various phases of the task. Surface EMG activity was recorded from three muscles contributing to wrist flexion and three muscles contributing to wrist extension. With negative stiffness, a significant compensatory increase in cocontraction of wrist flexor and extensor muscles was observed for slow movements, but there was little change in the muscle activity for rapid movements. With negative damping, muscle cocontraction was elevated to stabilize rapid movements, declining only gradually after the target was reached. For slow movements, cocontraction occurred only when negative damping was high. The response to square-wave vibration (10 Hz, +/-0.5 Nm), beginning at movement onset, was similar to that of negative damping, in that it resulted in elevated cocontraction. However, because the vibration persisted after the target was reached, there was no subsequent decrease in muscle activity. When the frequency was reduced to 5.5 Hz, but with the same torque impulse, cocontraction increased. This is consistent with greater mechanical instability. In summary, agonist antagonist cocontraction was adapted to the stability of the task. This generally resulted in less of a change in muscle activity during the movement phase, when the task was performed quickly compared with slowly. On the other hand, the change in muscle activity during stabilization depended more on the nature of the instability than the movement speed. PMID- 11914786 TI - Alterations in transport path differentially affect temporal and spatial movement parameters. AB - Many studies have examined the coordination of reach-to-grasp movements. However, there is debate regarding the mechanism of coordination between the transport and grasp components. The current study investigated the stability of temporal and spatial measures for reaches in which transport path was altered early or late in the reaching action. Transport alteration was accomplished by placing an obstacle either 10 cm (near) or 20 cm (far) from the hand starting position. Obstacle location affected the formation of transport path such that maximum wrist elevation coincided with the location of the obstacle. Kinematic analyses revealed that reaches over the near obstacle significantly prolonged transport time and time to maximum velocity compared with reaches over the far obstacle. A similar pattern of results was observed for the grasp component; reaches over the near obstacle resulted in a prolongation of grip duration, time to maximum aperture, and time to maximum opening and closing velocity. Grip closing velocity was decreased in the obstacle conditions. These results confirm findings from earlier studies that have shown that changes in the transport component affect grasp formation. A spatial and temporal analysis of grasp opening and closing was also performed. Grasp closing time varied significantly between conditions, while closing distance or the distance traveled by the wrist after maximum aperture remained essentially constant across conditions. These results suggest that the central nervous system may be using a spatial controller to coordinate prehensile components. PMID- 11914787 TI - Hemispheric asymmetry in visuospatial attention assessed with transcranial magnetic stimulation. AB - Transcranial magnetic stimulation (TMS) was used to study visuospatial attention processing in ten healthy volunteers. In a forced choice recognition task the subjects were confronted with two symbols simultaneously presented during 120 ms at random positions, one in the left and the other in the right visual field. The subject had to identify the presented pattern out of four possible combinations and to press the corresponding response key within 2 s. Double-pulse TMS (dTMS) with a 100-ms interstimulus interval (ISI) and an intensity of 80% of the stimulator output (corresponding to 110-120% of the motor threshold) was applied by a non-focal coil over the right or left posterior parietal cortex (PPC, corresponding to P3/P4 of the international 10-20 system) at different time intervals after onset of the visual stimulus (starting at 120 ms, 270 ms and 520 ms). Double-pulse TMS over the right PPC starting at 270 ms led to a significant increase in percentage of errors in the contralateral, left visual field (median: 23% with TMS vs 13% without TMS, P=0.0025). TMS applied earlier or later showed no effect. Furthermore, no significant increase in contra- or ipsilateral percentage of errors was found when the left parietal cortex was stimulated with the same timing. These data indicate that: (1) parietal influence on visuospatial attention is mainly controlled by the right lobe since the same stimulation over the left parietal cortex had no significant effect, and (2) there is a vulnerable time window to disturb this cortical process, since dTMS had a significant effect on the percentage of errors in the contralateral visual hemifield only when applied 270 ms after visual stimulus presentation. PMID- 11914788 TI - Cortical excitability and motor task in man: an investigation of the wrist extensor motor area. AB - Task-related changes in the corticospinal excitation of the right extensor carpi radialis (ECR) muscle were investigated in 16 healthy human subjects. The subjects were asked to perform a tonic isometric wrist extension or to clench their hand around a manipulandum, thereby coactivating the antagonistic wrist muscles. At matched levels of background EMG in the ECR muscle, transcranial magnetic stimulation (TMS) was applied through a figure-of-eight coil at 20-30 sites spaced 1 cm apart over the hand area of the left motor cortex. The cortical maps of the representation of the ECR muscle constructed in this way did not change between the two motor tasks. Nevertheless, for all investigated cortical sites TMS evoked a smaller motor evoked potential (MEP) in the ECR muscles during hand clenching than during wrist extension. A similar decrease in the short latency peak in the poststimulus time histogram (PSTH) of single ECR motor units to TMS during hand clenching was found in seven subjects (number of motor units = 35). In contrast, short-latency peaks in the PSTH evoked by electrical stimulation of the motor cortex had a similar size during the two tasks (number of motor units = 9; two subjects). Already the initial 0.5-1.0 ms of the short latency peak evoked by TMS was depressed during hand clenching, which suggests that decreased excitability of corticospinal cells with monosynaptic projections onto ECR motor units was involved. This decreased excitability was not explained by increased intracortical inhibition, which was found to be of a similar size during hand clenching and wrist extension. The task-related changes in the efficiency of the motor cortex output are discussed in relation to the function of the wrist antagonist muscles in handling and gripping tasks. PMID- 11914789 TI - Dorsal and ventral visual stream contributions to perception-action interactions during pointing. AB - The Ebbinghaus illusion, in which a central circle surrounded by large circles appears to be smaller than a central circle surrounded by small circles, affects the speed of pointing movements. When the central circle appears to be big, pointing movements directed towards it are faster than when the central circle appears to be small. This effect could be due to an interaction between ventral stream processing associated with determining relative object size and dorsal stream processing associated with sensorimotor output. Alternatively, the dorsal stream alone could mediate the effect via the transformation of object shape representations into motor output within the parietal lobe. Finally, ventral stream processing could be integrated into motor output through projections to the prefrontal cortex and subsequently to the motor areas of the cortex, thus bypassing the dorsal stream. These three alternatives were tested by disrupting either the ventral or dorsal stream processing using transcranial magnetic stimulation (TMS) while subjects made pointing movements as quickly and accurately as possible to the central target circles within the Ebbinghaus illusion display. The relative changes in reaction time, movement speed, and movement accuracy for small versus large appearing target circles were compared when TMS was delivered over each site as well as at a control site (SMA). The results showed that TMS over either the dorsal or ventral stream but not the SMA reduced the influence of the illusion on the pointing movement speed but did not affect reaction time or movement accuracy. A second control experiment was completed in which TMS was delivered during pointing movements to target circles of physically different sizes that were not surrounded by either large or small circles. This allowed us to determined whether the effect we observed in the main experiment was due specifically to the relative size information contained within the illusory display and the effect this has on the preparation of pointing responses or to an influence on basic perceptual and sensorimotor processes occurring within the ventral and dorsal streams, respectively. The results showed that the affect on pointing movement speed was still present with dorsal but not ventral stream stimulation. Taken together, this evidence suggests that the ventral stream contributes to pointing movements based on relative object size information via its projections to the prefrontal areas and not necessarily through interactions with the dorsal stream. PMID- 11914790 TI - Imagined and actual arm movements have similar durations when performed under different conditions of direction and mass. AB - Several experiments have suggested that similar physiological substrates are involved in movement execution and motor imagery, and that the same laws of movement control apply to both processes. Using a mental chronometry paradigm, we examined the effects of movement direction and added mass on the duration of actual and imagined movements. Six subjects executed or imagined arm movements in the sagittal and horizontal plane, in three different loading conditions: without added mass, and with an added mass of 1 and 1.5 kg. The duration of both actual and imagined movements was measured by an electronic stopwatch. The actual movements were significantly increased in duration as a function of mass, for both movement directions. However, direction per se had no effect on duration. The duration of imagined movements was very similar to that of actual movements whatever the subject and mass and direction condition. These results show that both inertial and gravitational constraints are accurately incorporated in the timing of the motor imagery process, which appears therefore to be functionally very close to the process of planning and performing the actual movement. PMID- 11914791 TI - Assessing the effect of posture change on tactile inhibition-of-return. AB - If a peripheral target follows an ipsilateral cue with a stimulus-onset asynchrony (SOA) of 300 ms or more, its detection is delayed compared to a contralateral-cue condition. This phenomena, known as inhibition-of-return (IOR), affects responses to visual, auditory, and tactile stimuli, and is thought to provide an index of exogenous shifts of spatial attention. The present study investigated whether tactile IOR occurs in a somatotopic vs an allocentric frame of reference. In experiment 1, tactile cue and target stimuli were presented to the index and middle fingers of either hand, with the hands positioned in an uncrossed posture (SOA 500 or 1,000 ms). Speeded target detection responses were slowest for targets presented from the cued finger, and were also slower for targets presented to the adjacent finger on the cued hand than to either finger on the uncued hand. The same pattern of results was also reported when the index and middle fingers of the two hands were interleaved on the midline (experiment 2), suggesting that the gradient of tactile IOR surrounding a cued body site is modulated by the somatotopic rather than by the allocentric distance between cue and target. PMID- 11914792 TI - Vestibular control of sympathetic activity. An otolith-sympathetic reflex in humans. AB - It has been proposed that a vestibular reflex originating in the otolith organs and other body graviceptors modulates sympathetic activity during changes in posture with regard to gravity. To test this hypothesis, we selectively stimulated otolith and body graviceptors sinusoidally along different head axes in the coronal plane with off-vertical axis rotation (OVAR) and recorded sympathetic efferent activity in the peroneal nerve (muscle sympathetic nerve activity, MSNA), blood pressure, heart rate, and respiratory rate. All parameters were entrained during OVAR at the frequency of rotation, with MSNA increasing in nose-up positions during forward linear acceleration and decreasing when nose down. MSNA was correlated closely with blood pressure when subjects were within +/-90 degrees of nose-down positions with a delay of 1.4 s, the normal latency of baroreflex-driven changes in MSNA. Thus, in the nose-down position, MSNA was probably driven by baroreflex afferents. In contrast, when subjects were within +/-45 degrees of the nose-up position, i.e., when positive linear acceleration was maximal along the naso-ocipital axis, MSNA was closely related to gravitational acceleration at a latency of 0.4 s. This delay is too short for MSNA changes to be mediated by the baroreflex, but it is compatible with the delay of a response originating in the vestibular system. We postulate that a vestibulosympathetic reflex, probably originating mainly in the otolith organs, contributes to blood pressure maintenance during forward linear acceleration. Because of its short latency, this reflex may be one of the earliest mechanisms to sustain blood pressure upon standing. PMID- 11914793 TI - Remapping of place cell firing patterns after maze rotations. AB - When place cells are recorded from rats running on an elevated T-maze inside a curtained enclosure that contains distinct, experimenter selected stimuli, rotations of the maze plus stimuli cause equal rotations of firing fields. Here, we examined the effects of conflicting rotations of a T-maze relative to a laboratory frame that contained a large number of fixed stimuli in the environment and asked whether positional firing patterns stayed in register with the maze or the room cues or were modified in some more complex way. After maze rotations of 90 degrees, 180 degrees or 270 degrees, firing fields were stable in the laboratory frame and thus shifted to a different maze arm. In contrast, rotations of 45 degrees or -45 degrees resulted in dramatic changes of positional firing patterns regardless of their initial position on the maze. Crucially, even cells whose fields were initially on the central platform underwent major firing pattern alterations although the view of the environment from the platform was unchanged by such rotations. Finally, we found that altering the visual appearance by removing without rotation one or two maze arms did not alter firing fields on the remaining part of the maze. Thus, the "remappings" caused by 45 degrees rotations could result from the disturbed relationship between all arms and the room cues or from the changes in the possible paths the animal can take in the environment. Taken together, our results provide an example of combinatorial coding by the hippocampus, in which the place cell representation of the environment was seen to be modified as a unit and not piecewise according to locally available stimuli. PMID- 11914794 TI - Redundant target effect and intersensory facilitation from visual-tactile interactions in simple reaction time. AB - In a simple reaction time (RT) task, normal observers responded faster to simultaneous visual and tactile stimuli than to single visual or tactile stimuli. RT to simultaneous visual and tactile stimuli was also faster than RT to simultaneous dual visual or tactile stimuli. The advantage for RT to combined visual-tactile stimuli over RT to the other types of stimulation could be accounted for by intersensory neural facilitation rather than by probability summation. The direction of gaze (and presumably of visual attention) to space regions near to or far from the site of tactile stimulation had no effect on tactile RT. However, RT to single or dual tactile stimuli was fastest when observers could see the sites of tactile stimulation on their hands both directly and through a mirror at the same time. All these effects can be ascribed to the convergence of tactile and visual inputs onto neural centers which contain flexible multimodal representations of body parts. PMID- 11914795 TI - Cortical activation associated with midtrial change of instruction in a saccade task. AB - The appearance of a visual stimulus in the peripheral visual field can elicit different saccade responses depending on prior instruction. This flexibility is commonly attributed to differences in motor set. Little is known about how the brain switches between one saccade response and another. To investigate the neural processes associated with switches between saccade motor sets, we recorded event-related potentials (ERPs) in 13 subjects, in three tasks that required subjects to generate prosaccades to a visual stimulus on 75% of the trials. On 25% of the trials, the color of the fixation point (FP) changed 300 ms prior to stimulus presentation. In the "ANTI" task, the change of the FP was the instruction to generate an antisaccade; in the "NOGO" task, subjects were instructed to maintain fixation; and in the "PRO" task, subjects were instructed to generate a prosaccade. The switch in motor set from prosaccades to antisaccades in the ANTI task and the cancellation of the prosaccade motor set in the NOGO task modulated frontal and frontocentral channels. Futhermore, the ANTI task but not the NOGO task was associated with differences at central and parietal channels compared with the PRO task. We hypothesize that the frontal activation in the ANTI and NOGO task reflects inhibition and task-switching processes, whereas the parietal activation reflects the preparation of this area for the sensorimotor transformation process that is necessary for the generation of an antisaccade. PMID- 11914796 TI - Histochemical identification of cortical areas in the auditory region of the human brain. AB - Despite numerous studies stretching over the last 100 years there is still no general agreement on the number of auditory areas in the human cortex or even how to define them by histological methods. Full definition of these areas will require a combination of functional and histological methods but, by using six complementary histological methods, of which most have been used in the monkey, we provide a clearer description of these areas. The primary auditory area was located on the posteromedial two-thirds of the first transverse temporal (Heschl's) gyrus and was distinguished by a dense band of cytochrome oxidase activity in layer IV and the base of layer III, as well as a relatively thick, pale layer V and VI. Layers V and VI together made up 40% of the cortical thickness. Acetylcholinesterase (AChE)-containing pyramidal cells were sparsely distributed within the primary auditory area. The anterolateral third of Heschl's gyrus did not have a clear band of high cytochrome oxidase activity but contained a moderately high density of AChE-containing pyramidal cells and thus appeared to be part of the auditory belt. Within Heschl's sulcus there was a third area, which had a band of high cytochrome oxidase activity and bands of high parvalbumin immunoreactivity and AChE activity in layer IV. This area appeared to be part of the auditory core. Thus the use of staining methods for cytochrome oxidase, AChE and parvalbumin provided additional information which allowed a clearer definition of auditory areas than Nissl or myelin staining alone. Our results suggest that there are two core areas surrounded by at least six belt areas in the human auditory region. PMID- 11914797 TI - Visual object memory and memory-guided saccades rely on shared mental representations. AB - We investigated the organisation of working memory processes by examining how a secondary memory task interferes with the accuracy of memory-guided saccades. A target was flashed at a random location, followed by a Kanji character. Subjects had to remember the location of the target and the Kanji character, and then they had to make a saccade towards the remembered target location. A second Kanji character was displayed and the subject had to decide if it was same or different. The performance of seven non-Kanji reader's were compared with six fluent Kanji readers in the task. Memorisation of Kanji characters was found to interfere with the accuracy of memory-guided saccades made by non-Kanji, but not by Kanji readers. These findings directly contradict accounts of working memory function which propose that spatial and visual object memory are functionally discrete. PMID- 11914798 TI - Contribution of geometry and joint stiffness to mechanical stability of the human arm. AB - This study investigates the ability to maintain a stable position of the hand when confronted with environmental instability. Subjects were required to hold the hand inside a 0.4-mm square while counteracting the destabilizing effect of a force field, which pushed the hand away from a line in the horizontal plane. The endpoint stiffness of the relaxed arm proved to be a reliable predictor of stability. Subjects were most successful in stabilizing hand position when the direction of the force field was aligned with the direction of greatest endpoint stiffness. They were least successful when the force field was aligned in the orthogonal direction, the direction of least endpoint stiffness. Subjects increased their endpoint stiffness as the strength of the force field was increased, but when the force field was in the direction of least stiffness they eventually failed in stabilizing the hand at the highest force field strength. In contrast, they were as successful in stabilizing the hand at the highest force field strength as at the lowest, when the force field was aligned with the direction of greatest stiffness. With the elbow flexed, endpoint stiffness of the relaxed arm becomes more uniform than with the elbow extended. This was reflected in subjects' performance, which improved considerably compared to the extended elbow posture, as force field strength was increased in the direction of lesser stiffness. The results indicate that posture was more effective than joint stiffness in stabilizing hand position. PMID- 11914799 TI - Does the motor control system use multiple models and context switching to cope with a variable environment? AB - Studies of arm movements have shown that subjects learn to compensate predictable mechanical perturbations by developing a representation of the relation between the state of motion of the arm and the perturbing forces. Here, we tested the hypothesis that subjects construct internal representations of two different force fields and switch between them when presented with an alternating sequence of these fields. Our results do not support this hypothesis. Subjects performed reaching movements in four sessions over 4 days. On the 1st day the robotic manipulandum perturbed the movement by perpendicular force that alternated its direction after each movement. Subjects were unable to construct the two underlying models and switch between them. On the 2nd day only one field was applied and well learned. On the 3rd day only the other field was applied and well learned. Then the experiment of the 1st day was repeated on the 4th day. Even after this extensive training subjects showed no signs of improved performance with alternating fields. This result combined with previous studies suggests that the central nervous system has a strong tendency to employ a single internal model when dealing with a sequence of perturbations. PMID- 11914800 TI - Directional control of reaching is preserved following mild/moderate stroke and stochastically constrained following severe stroke. AB - Recent evidence suggests that brain injury can impair the ability to independently activate shoulder and elbow muscles. We hypothesized that if muscle activation patterns are constrained, then brain-injured subjects should not be able to accurately grade initial hand movement direction during reaching toward a broad range of target directions. To test this hypothesis, we measured hand trajectories during reaching in three-space by 16 hemiparetic stroke subjects to an array of 75 targets distributed throughout the workspace. Contrary to our hypothesis, we found that the ability to grade movement direction was largely preserved following mild and moderate stroke. However, the most severely impaired subjects exhibited a degradation of directional control consistent with a loss of independent muscle control. Initial and final hand movement directions for these subjects were grouped roughly in two opposing directions, in a plane parallel with the coronal plane of the body, rather than distributed across the normal range. Selection between the two movement directions appeared partially random, in that subjects initiated over 50% of movements in the direction generally opposite the intended target, for targets to one side of the body. These results suggest that individuals with severe stroke are constrained to use only two gross, stereotypical muscle coactivation patterns for reaching control, and that selection between these patterns is stochastically influenced as the actual direction of motion is not strictly predictable given the desired direction. PMID- 11914802 TI - Semiquantitative dynamic computed tomography to predict response to anti-platelet therapy in acute cerebral infarction. AB - We investigated whether dynamic computed tomography (CT) in patients with acute cerebral infarction could identify patients likely to respond to anti-platelet therapy. Seventy patients underwent semiquantitative dynamic CT within 6 h as well as cerebral angiography. All then received anti-platelet therapy with a thromboxane A2 synthetase inhibitor. Peak value (pv) and time-to-peak (tp) (time density curves) for the Sylvian fissure were extracted from dynamic CT data and standardizing interpatient data, two indices, PV/TP index and TP index, were prepared following a standard semiquantitative manner. Both PV/TP index and TP index were effective in discriminating between 48 responders (modified Rankin scale (mRS): 0 to 2) and 22 non-responders (mRS: 3 to 5, or death: 6; both P<0.0001). High PV/TP index (>or=0.8) was a strong indicator of favorable response. Most of these patients maintained regional cerebral blood flow (rCBF) via anterograde flow or collaterals, with a TP index 1.1) and non-compensated rCBF. Intermediate PV/TP values could not predict outcome. Dynamic CT prior to therapy can identify patients with acute cerebral infarction who are treatable with anti-platelet therapy alone. PMID- 11914801 TI - Virtual endoscopy combined with intraoperative neuronavigation for planning of endoscopic surgery in patients with occlusive hydrocephalus and intracranial cysts. AB - We assessed the clinical value of MR ventriculoscopy (virtual endoscopy, VE) combined with image-guided frameless stereotaxy for endoscopic surgery of occlusive hydrocephalus and intracranial cysts. VE was obtained in 20 patients with hydrocephalus and three with intracranial cysts. All surgical operations were endoscopic. The path of the rigid endoscope to the target point was planned using neuronavigation. VE was carried out along the same trajectory retrospectively in 20 cases and prospectively in three. The results were analysed for demonstration of anatomical landmarks and structures at risk. VE was successful in all patients. Possible obstacles to endoscopic access to the lamina terminalis and the basal cisterns and structures at risk, such as the basilar artery, were clearly shown in relation to the direction of the endoscope. However, the floor of the third ventricle and septum pellucidum were not clearly seen and possible abnormalities could therefore not be appreciated. VE can provide realistic simulation of endoscopic third ventriculostomy and cystostomy. The appropriate trepanation point and trajectory of the endoscope can be assessed with regard to the size of the foramen of Monro and the position of vulnerable structures. This simulated trajectory can be adapted to the field of operation by image-guided neuronavigation. This regime may potentially reduce the risk of damage to intracranial structures. PMID- 11914803 TI - Serial analysis of the apparent diffusion coefficient time course in human stroke. AB - Acute cerebral ischemic injury can be rapidly detected on diffusion-weighted images. The apparent diffusion coefficient (ADC) depends on the stage of cytotoxic edema and water content in the infarcted parenchyma. The purpose of this study is to determine the time course of ADC during the first days of ischemic stroke. These data should make it possible to distinguish between multiple stroke and a single progressive infarction. Eight patients with clinically diagnosed acute cerebral ischemia were examined by diffusion-weighted MRI from 2 to 20 h after onset of symptoms. Daily control scans were performed for up to 10 days. ADC values were analyzed from 55 MRI studies. Furthermore, ADC was measured in the tissue which showed a hyperintense signal at the first examination and in the contralateral tissue. White and gray matter were analyzed separately. Data were expressed as the ratio ADC (rADC) of lesion to control region of interest. All patients showed a uniform reduction in rADC from the first hours of stroke and decreasing to the 3rd day. The rADC increased again from the 4th day up to the point of pseudo-normalization on day 9. The gray matter showed a slightly faster increase than the white matter. rADC shows significant changes in the first days after stroke, following a rather uniform time course. Together with T2-weighted MRI this makes it possible to differentiate between hyperacute, acute, and chronic stroke. Furthermore, the age of an ischemia can be determined and multiple strokes can be distinguished from a single progressive stroke. PMID- 11914804 TI - Serial investigation of perfusion disturbances and vasogenic oedema in hypertensive encephalopathy by diffusion and perfusion weighted imaging. AB - Serial MRI including diffusion and perfusion imaging was performed in a patient with hypertensive encephalopathy. At admission, the patient was disorientated and presented with seizures and cortical blindness. Perfusion imaging showed a marked reduction in blood volume and flow, with corresponding vasogenic oedema in the occipital, posterior temporal, and, to a lesser extent, frontal lobes. The clinical symptoms disappeared rapidly following treatment, whereas the disturbed circulation pattern and vasogenic oedema resolved more slowly. A complete normalisation was seen after 1 year. PMID- 11914805 TI - Transient decrease of water diffusion in Wernicke's encephalopathy. AB - We report water diffusion abnormalities in periventricular areas in a patient with Wernicke's encephalopathy. The reduction in diffusion disappeared after 2 weeks of treatment with intravenous thiamine. We suggest that the restricted mobility of cerebral water is related to inflammatory lesions. PMID- 11914806 TI - Early brain atrophy in HIV infection: a radiological-stereological study. AB - Patients with the acquired immunodeficiency syndrome (AIDS) frequently develop neurological abnormalities, and atrophy has been reported in neuropathological studies of brains from patients with AIDS. Our aim was to see whether increased ventricular volume, indicating central cerebral atrophy develops at an early stage of HIV-infection, before patients are severely immunodeficient. We also wished to follow ventricular size over time and to investigate whether enlargement would be more marked in patients, who became clinically demented. We studied 32 HIV-seropositive men who had several CT studies. At the time of the first CT the patients were divided into two groups, an immunocompetent group (IC) with CD4 cell count >400 and an immunodeficient group (ID) with CD4 cell count <400. These were compared with a control group of 44 seronegative men. Subsequent CT studies were used to measure changes in ventricular size with time. Ventricular volume was estimated with an unbiased stereological method previously applied to CT. Mean (geometric) ventricular volume was 15 ml (6-54 ml) in the controls, 23 ml (10-72 ml) in the IC group and 27 ml (8-80 ml) in the ID group. The increase in size in both patient groups was statistically significant compared with controls: P=0.005 (IC) and 9 x 10(-5)(ID). Regression analysis of the follow-up CT studies showed a significant increase in volume in the HIV group (r=0.74, P=0.002), but not in the AIDS group (r=0.41, P<0.1). PMID- 11914807 TI - High signal of the striatum in sporadic Creutzfeldt-Jakob disease: sequential change on T2-weighted MRI. AB - The object of this study is to describe the sequential change of high signal of the striatum on T2-weighted MRI in sporadic Creutzfeldt-Jakob disease (CJD). Three cases of autopsy-proven sporadic CJD and a total of 18 serial MR images are included in this study. The degree of high signal of the striatum on T2-weighted MRI was evaluated by two neuroradiologists and divided into four grades by mutual agreement. Initial MRI of all three cases showed a slightly high signal of the bilateral striatum, and the conspicuity of the high signal became more prominent as the disease progressed. In each case the pathological change of striatum and globus pallidus was compared with the high signal on the last MR image. PMID- 11914808 TI - Perimesencephalic nonaneurysmal subarachnoid haemorrhage and variations in the veins. AB - Perimesencephalic nonaneurysmal subarachnoid haemorrhage (PN-SAH) is characterised by accumulation of blood around the midbrain, normal angiographic findings and an excellent prognosis. The etiology of PN-SAH has not yet been determined. Therefore we decided to compare the venograms of patients with PN-SAH with those of patients with aneurysmal SAH (A-SAH) in order to examine the relationship between PN-SAH and venous structures. We retrospectively studied 6 patients with PN-SAH and 102 cases of angiographically evaluated A-SAH during the past 12 years by reviewing their venograms for possible abnormalities in venous structures, particularly in the basal vein of Rosenthal (BVR). More abnormalities in venous structures were found in the patients with PN-SAH than in those with A SAH. Most of the BVR in the patients with PN-SAH appeared to drain into various dural sinuses instead of the galenic system. The relationship between PN-SAH and abnormalities in venous structures was determined. PMID- 11914809 TI - Apparent diffusion coefficient measurements within intracranial epidermoid cysts in six patients. AB - The purpose was to determine whether a strong decrease in apparent diffusion coefficient (ADC) within epidermoid cysts (ECs) is actually responsible for their bright signal intensity on diffusion-weighted (DW) trace images. We studied six patients with surgically proven ECs in whom ADC calculation from T2-weighted DW EPI-SE data were performed within the ECs and within the deep white matter and cerebrospinal fluid (CSF) as references. All ECs displayed highest signal intensity on the DW trace images. ADC values ranged from 1,280 to 807 x 10(-6) mm(2)/s within cysts (with a mean value of 1,070), from 849 to 698 x 10(-6) mm(2)/s within white matter (with a mean value of 764) and from 3,370 to 2,980 x 10(-6) mm(2)/s within CSF (with a mean value of 3,185). ECs exhibited slightly higher ADC values than white matter, and not the strongly decreased ones which would have been expected if diffusion-weighting were the prominent mechanism for bright signal intensity of the ECs on DW images. However, the EC ADC values are much lower than those of the CSF. Other mechanisms must therefore be involved, i.e. the T2 shine-through effect. Reduced ADC is not the only explanation of the EC bright signal intensity on the DW trace images. PMID- 11914810 TI - Ganglioglioma causing acute uncal herniation. AB - Gangliogliomas are rare but mostly benign tumors of the central nervous system. Early surgical resection is recommended to prevent seizures and further growth. Therefore, an uncal herniation caused by a ganglioglioma is a very rare event. We report such a case, where a patient with an initially small lesion was lost to follow-up. PMID- 11914811 TI - Neuroradiological findings in two cases of isolated amyloidoma of the central nervous system. AB - Neuroradiological findings in two cases of isolated cerebral amyloidoma are reported. Case 1: A 15-year-old male admitted because of left jacksonian seizure. Case 2: A 31-year-old man admitted because of temporal seizures. In both patients, CT scans showed spontaneously hyperdense and enhancing nodules. On MRI, lesions displayed signals isointense with surrounding edema on T1-weighted sequences and mildly hypointense on T2-weighted sequences if compared with edema; peripheral contrast enhancement after gadolinium administration was present. Histopathological examination of both specimens revealed the presence of acellular eosinophilic hyalinized material consistent with amyloid, forming confluent nodular masses in the white and gray matter. PMID- 11914812 TI - Primary Sjogren's syndrome initially manifested by optic neuritis: MRI findings. AB - We herein describe the MRI findings in a patient clinically diagnosed with primary Sjogren's syndrome (SjS) initially manifested by retrobulbar optic neuritis. A 63-year-old woman suddenly had left ocular pain and progressive visual disturbance. MR T2-weighted images revealed hyperintensity in the left optic nerve, with swelling. Contrast-enhanced T1-weighted images showed no abnormal enhancement. Follow-up MRI 6 months after admission revealed no significant changes in the affected optic nerve. To our knowledge, optic neuritis as a complication of SjS has been reported in ten patients [1, 2, 3, 4, 5, 6] and MRI findings in only one of them [6]. We thought MR images were useful for visualizing optic nerve involvement in SjS and observing its course. PMID- 11914813 TI - Venous angioma adjacent to the root entry zone of the trigeminal nerve: implications for management of trigeminal neuralgia. AB - Detection of a venous angioma at the root entry zone is important for surgical planning, so that the neurosurgeon will be aware that both veins and arteries may require microvascular decompression. In selected cases, alternative treatment may be indicated to avoid the potential surgical complication of a venous infarct. Trigeminal neuralgia typically occurs in the middle-aged to elderly population, usually the result of compression of the trigeminal nerve at its root entry zone by an ectatic, aging artery or, less commonly, a regional vein [1, 2, 3]. When associated with a venous angioma at the root entry zone, trigeminal neuralgia usually presents at a younger age [4, 5, 6]. We review the imaging examinations and clinical data of five patients with trigeminal neuralgia who had a venous angioma adjacent to the root entry zone of the trigeminal nerve, and discuss how the imaging findings affected their management. PMID- 11914814 TI - Potential role of the anterior spinal artery in preventing propagation of thrombus in a therapeutically occluded vertebral artery: angiographic studies before and after endovascular treatment. AB - Therapeutic occlusion of the vertebral artery (VA) is one of the treatments for unclippable aneurysms and other lesions, although controversy still surrounds the appropriate site for occlusion to attain selective thrombosis of the lesion while avoiding ischaemic complications. The lower two-thirds of the lateral medulla are supplied by perforating branches of both the VA and the posterior inferior cerebellar artery (PICA). However, in patients without a PICA or in whom the origin of the PICA is low (at or below the foramen magnum), the VA is usually the only source of perforating vessels. We retrospectively studied the results of VA occlusion on such anatomically high-risk patients, and propose a safer procedure. Five high-risk patients underwent therapeutic occlusion of the VA for dissecting aneurysms or arteriovenous fistula. A lateral medullary syndrome developed due to propagation of thrombus after the procedure in two patients in whom angiography did not demonstrate the anterior spinal artery (ASA) within the stump of the VA. Ischaemic signs did not develop in the other three patients, in whom the ASA was visible, and retrograde flow was observed proximal to the origin of the ASA. This suggests that the ASA may play a role in preventing propagation of thrombus in the VA distal to the site of occlusion and supply blood to its perforating arteries in high-risk patients. Angiographic assessment of the ASA may be useful for predicting the likelihood of the lateral medullary syndrome developing with therapeutic occlusion of the VA in patients without a PICA or with one whose origin is low. PMID- 11914816 TI - A high-resolution MRI study of linear growth of the human fetal skull base. AB - The skull base, otherwise referred to as the basicranium or cranial base, plays a key role in the process of skull development, providing both support for the brain and an architectural component of the craniofacial complex. Consequently, the fetal skull base has been the focus of numerous studies employing various methods, including sectioning, plain radiography and CT. This paper investigates high-resolution (hr) MRI as an alternative method for looking at and quantifying the fetal skull base. The evaluation tests two basic hypotheses drawn from previous studies. These suggest that the anterior segment of the midline skull base grows more rapidly than the posterior segment and that the width of the posterior cranial fossa increases disproportionately in relation to its length. I imaged 42 formalin preserved human fetuses from museum collections with hrMRI. The T2-weighted image voxels were significantly smaller than those acquired with conventional clinical MRI. Landmarks of the fetal skull base were identified on reformatted axial and sagittal images. Bivariate plots revealed that the growth rate of the anterior skull base is almost twice that of the posterior skull base and that increases in the width of the posterior cranial fossa exceed those in its length. These findings confirm those of previous investigations and show that hrMRI offers a way forward in noninvasive quantification of fetal morphology. PMID- 11914815 TI - Thromboembolic events after endovascular treatment of unruptured intracranial aneurysms in two patients with antiphospholipid-antibody syndrome. AB - Antiphospholipid antibodies (APAs) are circulating immunoglobulins associated with a hypercoagulable state. The antiphospholipid syndrome combines APAs and clinical manifestations, including arterial or venous thromboses and/or recurrent spontaneous fetal loss. The main risk incurred by endovascular treatment of intracranial aneurysms is the occurrence of thromboembolic events. We report two cases of patients with antiphospholipid syndrome who developed thromboembolic complications after the endovascular treatment of unruptured intracranial aneurysms. PMID- 11914880 TI - FDG PET in the management of lymphoma: a clinical perspective. PMID- 11914881 TI - Feasibility, safety and image quality of cardiac FDG studies during hyperinsulinaemic-euglycaemic clamping. AB - Fluorine-18 fluorodeoxyglucose (FDG) imaging for the assessment of myocardial viability has become an integral part of the diagnostic and prognostic work-up of patients with ischaemic cardiomyopathy. To ensure good image quality, in particular in patients with diabetes mellitus, hyperinsulinaemic-euglycaemic clamping has been proposed. In this study we evaluated the safety and the image quality of cardiac FDG imaging during clamping in a large group of patients, including a subgroup with diabetes mellitus. The incidence of viability (on both a segment and a patient basis) was also determined for patients with and without diabetes mellitus. The safety and image quality of cardiac FDG studies during clamping were evaluated in 131 patients, including 19 with diabetes mellitus. Image quality was assessed visually and quantitatively using heart-to-lung (H/L), heart-to-liver (H/Li) and myocardium-to-background (M/B) ratios. Blood samples were drawn at baseline and at the time of FDG injection to determine levels of glucose, free fatty acids and insulin. The metabolic circumstances were optimal for FDG imaging: high insulin levels, low free fatty acid levels and glucose levels in the normal range (levels of substrates were comparable between patients with and patients without diabetes mellitus). No serious side-effects occurred in any patient. Image quality (assessed visually) was good in all patients. The quantitative parameters of image quality (H/L, H/Li and M/B) were comparable between patients with and patients without diabetes mellitus. The incidence of viability was high: 38% of patients without and 58% of patients with diabetes mellitus had substantial viability despite contractile dysfunction. It is concluded that cardiac FDG imaging during clamping is safe and provides excellent image quality, including in patients with diabetes mellitus. The incidence of viability is high, in particular in patients with diabetes mellitus. PMID- 11914882 TI - Early prediction of regional functional recovery in reperfused myocardium using single-injection resting quantitative electrocardiographic gated SPET. AB - By evaluating concordant or discordant perfusion and systolic wall thickening patterns, resting quantitative electrocardiographic (ECG) gated single-photon emission tomography (SPET) can identify various myocardial pathological conditions with different functional recovery after revascularisation therapy. However, no data are available on the ability of this methodology to predict regional functional recovery after primary percutaneous transluminal coronary angioplasty (PTCA). This study evaluated whether single-injection ECG gated SPET imaging performed at rest with 99mTc-tetrofosmin early after successful PTCA can predict recovery of regional wall motion. ECG gated SPET was performed 3 days and 3 weeks after successful PTCA in 26 patients. Regional functional parameters were automatically calculated with a 20-segment model on the day 3 image, and segments with perfusion/thickening mismatch were defined as showing preserved perfusion (>55% uptake on the end-diastolic image: mean-standard deviation of the normal value) without systolic wall thickening (mean-standard deviation of the normal value). On the third day, the regional wall motion score of 37 mismatched segments (3.8+/-2.1) was significantly lower than that of 41 matched normal segments (6.0+/-2.9), but was significantly higher than that of 108 matched abnormal segments (1.4+/-1.9, both P<0.01). At 3 weeks after acute MI, the regional wall motion score of mismatched segments (6.4+/-3.9) improved to the level of matched normal segments (7.1+/-3.0) and was significantly higher than that of matched abnormal segments (2.5+/-3.0, P<0.01). Absolute change in the regional wall motion score (3 days to 3 weeks) of mismatched segments (2.6+/-3.5) was significantly greater than that in the regional wall motion score of matched normal segments and matched abnormal segments (1.1+/-1.3 and 1.2+/-2.6, respectively, both P<0.05). Twenty-seven of 37 segments (73%) with perfusion/thickening mismatch showed significant improvement in regional wall motion, whereas improvement in regional wall motion was observed in 22 of 108 segments (20%) with matched abnormal segments and 6 of 41 segments (15%) with matched normal segments. Segments with perfusion/thickening mismatch had a significantly higher incidence of regional functional improvement than did matched abnormal or matched normal segments (chi2=42.3, P<0.01). Thus, by estimating both perfusion and wall thickening, single-injection resting ECG gated SPET imaging with 99mTc-tetrofosmin early after primary PTCA can predict recovery of regional wall motion after successful reperfusion. PMID- 11914883 TI - Reconstruction of gated myocardial perfusion SPET incorporating temporal information during iterative reconstruction. AB - Reconstruction of gated single-photon emission tomography (gSPET) is intrinsically a four-dimensional (4D) problem. In practice, the time frames are reconstructed independently as a sequence of frame-by-frame reconstructions. This approach is not optimal since the strong signal correlations among the individual time frames are not exploited. In this study we propose a simple but efficient algorithm to improve the image quality of myocardial perfusion gSPET by incorporating the cyclic temporal information within the reconstruction using Fourier filtering. The gSPET images were reconstructed using the Ordered Subsets Expectation Maximisation (OSEM) algorithm employing six iterations with eight subsets. Temporal filtering was applied either before (PreOSEM) or after image reconstruction (PostOSEM) or was incorporated within the OSEM algorithm (OSEM4D). The effect of temporal filtering was compared with conventional frame-by-frame OSEM using clinical data. Image quality was evaluated by estimating the systematic and statistical error. The results indicated that temporal filtering introduces a small (<1%) systematic error, while the statistical error was reduced from 15.0%+/-3.1% when conventional frame-by-frame OSEM was applied to 12.6%+/-2.7%, 12.0%+/-2.5% and 9.3%+/-2.4% when PreOSEM, PostOSEM and OSEM4D were used, respectively. It is concluded that temporal filtering incorporated within OSEM reconstruction dramatically reduces noise in gated SPET myocardial images. PMID- 11914884 TI - The usefulness of hepatobiliary scintigraphy in the diagnosis of complications after adult-to-adult living donor liver transplantation. AB - Living donor liver transplantation has become an accepted procedure to overcome the shortage of adult donor organs. The aim of this study was to evaluate the usefulness of hepatobiliary scintigraphy in the diagnosis of complications after adult-to-adult living donor liver transplantation. We analysed 82 hepatobiliary scintigraphy studies performed using technetium-99m DISIDA in 60 adult patients (44 males, 16 females) who had been transplanted with a living donor's hepatic lobe (right lobe, 32; left lobe, 28). Indications for hepatobiliary scintigraphy were abnormal symptoms and/or liver function tests ( n=54) or suspected bile leak or biloma ( n=28). Median interval between transplantation and scintigraphy was 69 days (9 days to 23 months). Scintigraphic findings were classified into hepatic parenchymal dysfunction, total biliary obstruction, segmental biliary obstruction, bile leak and normal graft. Scintigraphic findings were confirmed by liver biopsy in 17 cases, and by radiological and clinical follow-up in 65 cases. There were 29 events relating to biliary complications (six total biliary obstructions, eight segmental biliary obstructions and 15 bile leaks) and 19 relating to non-biliary complications (15 cases of rejection, two of infection and two of vascular compromise) in 38 patients. Hepatobiliary scintigraphy provided the correct diagnosis in all eight segmental and five of six total biliary obstructions, and in all 15 cases of bile leak. Of the 19 non-biliary complications, 16 showed parenchymal dysfunction regardless of the aetiology and three showed total biliary obstruction on scintigraphy. All but three of 34 normally functioning grafts were normal on scintigraphy. The diagnostic sensitivity and specificity of scintigraphy for biliary obstruction in the 54 patients with abnormal symptoms or liver function tests were 93% (100% for segmental, 83% for total) and 88% (35/40), respectively. The sensitivity and specificity were each 100% (15/15, 13/13) for bile leak in the 28 patients with suspected bile leak or biloma. Hepatobiliary scintigraphy is an accurate diagnostic modality in the evaluation of biliary complications after adult-to adult living donor liver transplantation, although it has limitations as a means of differential diagnosis of non-biliary complications. PMID- 11914885 TI - Dose selection for radioiodine therapy of borderline hyperthyroid patients with multifocal and disseminated autonomy on the basis of 99mTc-pertechnetate thyroid uptake. AB - The aim of this study was to optimise radioiodine therapy of diffuse and nodular toxic goitre by calculation of the radiation dose delivered to the thyroid on the basis of the pretreatment technetium-99m pertechnetate thyroid uptake under thyrotropin suppression (TcTU(s)). The TcTU(s) value serves as a substitute for the non-suppressible iodine turnover and the functional autonomous mass. Marinelli's formula was used to calculate tissue absorbed doses of 150 Gy, 200 Gy, 250 Gy and 300 Gy to the thyroids of 438 patients with multifocal and disseminated autonomy. The mean age of patients was 70+/-9 years, and the mean thyroid volume was 54+/-26 ml. Two hundred and sixty-one of the patients had at least one documented previous episode of overt hyperthyroidism. Tissue absorbed doses were adapted to the pretreatment TcTU(s): 150 Gy for a TcTU(s) of 1.5% 2.49%, 200 Gy for a TcTU(s) of 2.5%-3.49%, 250 Gy for a TcTU(s) of 3.5%-4.49% and 300 Gy for a TcTU(s) of > or =4.5%. Normalisation of TcTU(s) and thyrotropin (TSH), thyroid volume reduction and frequency of hypothyroidism and recurrent hyperthyroidism were evaluated 1 year after a single radioiodine therapy. The presented dose strategy resulted in normalisation of TcTU(s) in 96% and an increase in TSH to the normal range in 92%. Recurrent hyperthyroidism was observed in only five patients. Thyroid volume decreased from 54+/-26 before treatment to 34+/-20 ml, a mean reduction of 37%. The frequency of hypothyroidism, at 0.9%, was encouragingly low. Dose selection in accordance with pretreatment TcTU(s) can be recommended for elimination of functional autonomous tissue with a single radioiodine therapy in patients of advanced age with enlarged thyroid glands and relevant autonomous masses who are at risk of developing iodine-induced hyperthyroidism. PMID- 11914886 TI - Local injection of the 90Y-labelled peptidic vector DOTATOC to control gliomas of WHO grades II and III: an extended pilot study. AB - We have previously presented preliminary observations on targeting somatostatin receptor-positive malignant gliomas of all grades by local injection of the radiolabelled peptidic vector 90Y-DOTATOC. We now report on our more thorough clinical experience with this novel compound, focussing on low-grade and anaplastic gliomas. Small peptidic vectors have the potential to target invisible infiltrative disease within normal surrounding brain tissue, thereby opening a window of opportunity for early intervention. Five progressive gliomas of WHO grades II and III and five extensively debulked low-grade gliomas were treated with varying fractions of 90Y-DOTATOC. The vectors were locally injected into the resection cavity or into solid tumour. The activity per single injection ranged from 555 to 1,875 MBq, and the cumulative activity from 555 to 7,030 MBq, according to tumour volumes and eloquence of the affected brain area, yielding dose estimates from 76+/-15 to 312+/-62 Gy. Response was assessed by the clinical status, by steroid dependence and, every 4-6 months, by magnetic resonance imaging and fluorine-18 fluorodeoxyglucose positron emission tomography. In the five progressive gliomas, lasting responses were obtained for at least 13-45 months without the need for steroids. Radiopeptide brachytherapy had been the only modality applied to counter tumour progression. Interestingly, we observed the slow transformation of a solid, primarily inoperable anaplastic astrocytoma into a resectable multi-cystic lesion 2 years after radiopeptide brachytherapy. Based on these observations, we also assessed the feasibility of local radiotherapy following extensive debulking, which was well tolerated. Targeted beta-particle irradiation based on diffusible small peptidic vectors appears to be a promising modality for the treatment of malignant gliomas. PMID- 11914887 TI - Strontium-89 (Metastron) and the bisphosphonate olpadronate reduce the incidence of spinal cord compression in patients with hormone-refractory prostate cancer metastatic to the skeleton. AB - Spinal cord compression (SCC) is a devastating complication of metastatic cancer. We investigated the potential beneficial effect of two palliative therapies- strontium-89 (Metastron) and the nitrogen-containing bisphosphonate olpadronate- on the incidence of SCC in hormone-refractory prostate cancer (HRPC) metastatic to the skeleton. We retrospectively studied 415 patients with histologically proven prostate cancer who underwent bone scintigraphy at the time of diagnosis and were followed up at the Leiden University Medical Center between 1990 and 1999. Medical or surgical castration was undertaken in 172 patients with evidence for skeletal metastases. Within 2 years, 147 of these patients (85%) developed HRPC associated with severe progressive bone pain. Palliative treatment was given to 131 patients in the form of local radiotherapy ( n=10), 89Sr ( n=46) or intravenous olpadronate ( n=66), with ( n=57) or without ( n=9) maintenance oral olpadronate. Nine patients received both 89Sr and olpadronate at various intervals. Sixteen patients who did not receive any of these treatments were used as historical controls. There was no significant difference in baseline characteristics between treatment modalities. The incidence of SCC was 17% in the whole group, and highest in controls receiving no palliation (50%). None of the patients treated with local radiotherapy, only 4% of patients receiving 89Sr and 21% of patients given olpadronate developed this complication. Our findings suggest a significant reduction in SCC in patients with symptomatic HRPC metastatic to the skeleton who receive palliative therapies. Local radiotherapy completely prevents the incidence of SCC, 89Sr leads to an important decrease in this complication and olpadronate induces a significant, albeit smaller decrease in the incidence of SCC. The use of these agents opens new avenues in the difficult management of patients with advanced prostate cancer who are most at risk of developing SCC. PMID- 11914888 TI - Unlabelled iododeoxyuridine increases the rate of uptake of [125I]iododeoxyuridine in human xenografted glioblastomas. AB - 5-Iodo-2'-deoxyuridine (IdUrd), a thymidine (TdR) analogue, can be radiolabelled with iodine-125, an Auger radiation emitter, to provoke double-strand breaks once incorporated into DNA of cancer cells. We have previously shown that co incubation of [125I]IdUrd with unlabelled IdUrd provided an additive cytotoxicity in two human glioblastoma cell lines. This observation was unexpectedly correlated with an increase in the rate of DNA incorporation of [125I]IdUrd. Here, we further evaluated the effects of unlabelled IdUrd on the uptake of [125I]IdUrd in vitro and in vivo in mice xenografted with three human glioblastoma lines. The results showed that, in these three glioblastoma lines, unlabelled IdUrd increased the rate of uptake of [125I]IdUrd in vitro by 2- to 4.4-fold and in vivo by 1.5- to 2.8-fold. The rate of uptake of [125I]IdUrd in normal rapidly dividing tissues was also increased by 1.3- to 2.8-fold. TdR completely blocked [125I]IdUrd uptake in tumours and tissues. Analogues of IdUrd, such as deoxyuridine and 5-iodo-1,3-dimethyuracil, did not reproduce the effect of IdUrd on the uptake of [125I]IdUrd, suggesting that it is not related to protection against [125I]IdUrd degradation. It is concluded that combined administration of unlabelled IdUrd may improve the use of radiolabelled IdUrd for cancer diagnosis or therapy. PMID- 11914889 TI - Evaluation of high-risk melanoma: comparison of [18F]FDG PET and high-dose 67Ga SPET. AB - Recently the potential of whole-body positron emission tomography scanning using 18F-fluorodeoxyglucose (FDG PET) has led to renewed interest in the use of functional imaging for the detection of occult metastatic melanoma. This study compared dedicated FDG PET with high-dose gallium-67 imaging incorporating whole body scanning and comprehensive single-photon emission tomography (SPET) in 122 cases (121 patients) in which the two scans were performed <6 weeks apart. All patients were at high clinical risk of occult metastatic disease and 49 (40%) had abnormality suggestive of metastatic disease by at least one functional imaging technique. Discrepant scan findings were followed up to determine which technique more accurately reflected disease status. There were 23/122 (19%; 95% CI: 12% 26%) cases with discordant scan results in respect of either the presence of melanoma (11 cases) or the extent of disease (12 cases). PET correctly identified more disease than 67Ga SPET in 14 cases (including three incidental primary tumours) and was true negative in three further patients with abnormal 67Ga SPET. There were six patients with true positive 67Ga SPET in whom FDG PET was false negative (one small cutaneous deposit, one residual axillary node rated equivocal on FDG PET due to postoperative changes, one adrenal metastasis inseparable from renal activity on FDG PET and three cases in which sites missed on FDG PET were seen on 67Ga SPET. Thus, FDG PET provided incremental diagnostic information compared with 67Ga SPET in 17/23 patients, while 67Ga SPET provided incremental information compared with PET in 6/23 cases ( P=0.035). Based on Australian Medicare reimbursement levels, the net cost per patient with clinical management benefit of replacing 67Ga SPET with FDG PET was estimated to be less than EUR 1,750. These results suggest that FDG PET provides incremental and clinically important information in around 10% of patients at a low incremental cost which, combined with greater patient convenience and lower radiation dosimetry, make FDG PET the functional imaging technique of choice for evaluation of suspected metastatic melanoma. PMID- 11914890 TI - 18F-FDG hybrid PET in patients with suspected spondylitis. AB - This study investigated the value of fluorine-18 2'-deoxy-2-fluoro- D-glucose (FDG) imaging with a double-headed gamma camera operated in coincidence (hybrid PET) detection mode in patients with suspected spondylitis. Comparison was made with conventional nuclear medicine imaging modalities and magnetic resonance imaging (MRI). Sixteen patients with suspected spondylitis (nine male, seven female, mean age 59 years) prospectively underwent FDG hybrid PET (296 MBq) and MRI. For intra-individual comparison, the patients were also imaged with technetium-99m methylene diphosphonate (MDP) (555 MBq) ( n=13) and/or gallium-67 citrate (185 MBq) ( n=11). For FDG hybrid PET, two or three transverse scans were performed. Ratios of infected (target) to non-infected (background) (T/B) vertebral bodies were calculated. MR images were obtained of the region of interest. Patients found positive for spondylitis with MRI and/or FDG hybrid PET underwent surgical intervention and histological grading of the individual infected foci. Twelve out of 16 patients were found to be positive for spondylitis. Independent of the grade of infection and the location in the spine, all known infected vertebrae ( n=23, 9 thoracic, 12 lumbar, 2 sacral) were detected by FDG hybrid PET. T/B ratios higher than 1.45+/-0.05 (at 1 h p.i.) were indicative of infectious disease, whereas ratios below this value were found in cases of degenerative change. FDG hybrid PET was superior to MRI in patients who had a history of surgery and suffered from a high-grade infection in combination with paravertebral abscess formation ( n=2; further computed tomography was needed) and in those with low-grade spondylitis ( n=2, no oedema) or discitis ( n=2, mild oedema). False-positive 67Ga citrate images ( n=5: 2 spondylodiscitis, 1 aortitis, 1 pleuritis, 1 pulmonary tuberculosis) and 99mTc-MDP SPET ( n=4: 1 osteoporosis, 2 spondylodiscitis, 1 fracture) were equally well detected by FDG hybrid PET and MRI. No diagnostic problems were seen in the other patients ( n=5). In this study, FDG hybrid PET was superior to MRI, 67Ga citrate and (99m)Tc MDP, especially in patients with low-grade spondylitis (as compared with MRI), adjacent soft tissue infections (as compared with 67Ga citrate) and advanced bone degeneration (as compared with 99mTc-MDP). PMID- 11914891 TI - Whole-body PET with FDG for the diagnosis of recurrent gastric cancer. AB - This retrospective study was designed to assess the accuracy of fluorine-18 fluorodeoxyglucose positron emission tomography (FDG-PET) in diagnosing recurrence of gastric cancer. Thirty-three patients who had received surgical treatment for gastric cancer with curative intent and who had subsequently undergone FDG-PET for suspected recurrence were retrieved from the PET database. All patients were reviewed with full knowledge of prior conventional diagnostic work-up. Results were compared with a gold standard, consisting of histological confirmation or radiological and clinical follow-up. The gold standard established disease recurrence in 20/33 patients (prevalence 61%). Sensitivity and specificity of FDG-PET for the diagnosis of recurrence were 70% (14/20) and 69% (9/13), respectively. Positive and negative predictive values were 78% (14/18) and 60% (9/15), respectively. Of the six false-negative cases, all had intra-abdominal lesions (three had generalised abdominal metastases, one liver metastasis, one local recurrence and one ovarian metastasis). In the subgroup with previous signet cell differentiation of the primary tumour ( n=13, disease prevalence 62%), sensitivity was 62% (5/8) and specificity, 60% (3/5). Survival analysis for the entire patient group using Kaplan-Meier statistics yielded a longer survival in the PET-negative group (mean+/-SD, 21.9+/-19.0 months) than in the PET-positive group (mean+/-SD, 9.2+/-8.2 months) ( P=0.01). In the patient group with proven recurrence ( n=20), the mean survival for the PET-negative group was 18.5 (+/-12.5) months, as compared with 6.9 (+/-6.5) months for the PET positive group ( P=0.05). Because of its poor sensitivity and low negative predictive value, FDG-PET is not suited for screening purposes in the follow-up of treated gastric cancer. However, FDG-PET appears to provide important additional information concerning the prognosis of recurrent gastric cancer. PMID- 11914892 TI - Infecton is not specific for bacterial osteo-articular infective pathology. AB - The aim of this study was to re-examine, by retrospective analysis of our case material, the specificity and sensitivity of technetium-99m ciprofloxacin scan in discriminating between infection and other conditions. (99m)Tc-ciprofloxacin scintigraphy was performed in 71 patients: 30 patients referred for suspicion of osteomyelitis (OM) or septic arthritis (SA) (group 1) and 41 controls (group 2). Imaging was performed at 4 h post injection and, when possible, at 8 or 24 h post injection. Tracer uptake was visually assessed in different joint groups, and in the sites suspicious for infection. Several soft tissue sites were also evaluated. In the group referred for osteo-articular infection, we found a lower specificity (54.5%) than has previously been reported in the literature. Evaluation of tracer uptake at late imaging did not improve discrimination between sterile and non-sterile inflammation. Additionally, articular uptake was seen in many control patients. Infecton uptake in growth cartilage, thyroid gland, vascular pool, lungs, liver and intestines is discussed. PMID- 11914893 TI - Scintigraphic follow-up of the effects of therapy with hydroxyurea on splenic function in patients with sickle cell disease. AB - Patients with sickle cell disease (SCD) may develop functional asplenia as a chronic complication, secondary to repeated episodes of polymerisation of haemoglobin S. It is known that increased plasma concentrations of fetal haemoglobin (HbF) reduce the polymerisation of haemoglobin S. Hydroxyurea is a chemotherapeutic agent capable of increasing HbF levels in the red blood cells and its use has recently been proposed in the treatment of SCD. The objective of this study was to evaluate the effects of long-term therapy with hydroxyurea on recovery of splenic function. Twenty-one patients (aged 3-22 years; 14 with SS haemoglobinopathy, 7 with Sbeta(0) haemoglobinopathy) were studied with liver/spleen scintigraphy before and after 6 and 12 months of treatment. All studies were submitted to visual inspection and semi-quantitative analyses using spleen/liver ratios. Imaging prior to treatment demonstrated functional asplenia in nine SS patients and one Sbeta(0) patient and impaired splenic function in five SS patients and six Sbeta(0) patients. After treatment, splenic function improved in ten patients, remained unchanged in eight and worsened in three. Using liver/spleen imaging, it was possible to demonstrate that hydroxyurea is capable of improving splenic function in some SCD patients. Improvement is not always possible and frequently does not lead to a normal splenic function even after 1 year of treatment. PMID- 11914894 TI - FDG-PET as a "metabolic biopsy" tool in non-lung lesions with indeterminate biopsy. AB - The differentiation of benign versus malignant disease in a lesion identified on conventional imaging is a commonly encountered problem. Attempted biopsy is often unsuccessful or falsely reassuring and may lead to the patient being sent for more invasive and potentially morbid investigations. Having previously identified the value of fluorine-18 fluorodeoxyglucose positron emission tomography (FDG PET) in this circumstance in patients with lung lesions, our current aim was to investigate the role of FDG-PET in helping to identify more accurately those patients with malignant lesions outside the lung. FDG-PET scanning was performed in 50 patients; most had undergone unsuccessful biopsy of a lesion outside the lung, while in a smaller number no attempt at biopsy had been made as it had been considered too dangerous. Follow-up was by histology or, if this was unavailable, by clinical progress to death or a minimum of 12 months post scan. Visual and quantitative analysis was performed. On visual analysis, the positive and negative predictive values were 89% and 100%, respectively. On quantitative (SUV>2.5) analysis, positive and negative predictive values were 93% and 86%, respectively. A negative FDG-PET study in these circumstances virtually excludes malignancy and allows the patient to be reassured. A positive scan encourages the clinician to pursue further biopsy to confirm a histological diagnosis. FDG-PET therefore assists in deciding which patients need to undergo further investigation. PMID- 11914897 TI - Nuclear cardiology in the UK: activity and practice 1997. AB - A questionnaire was sent to 251 nuclear medicine centres asking for details of nuclear medicine activity, and nuclear cardiology activity and practice in 1997. One hundred and seventy-one (68%) centres replied. Nuclear medicine activity was estimated at 11.8 studies/1,000 population/year, and 9.5% of these studies were within cardiology (1.12 studies/1,000/year). Myocardial perfusion imaging (MPI) studies accounted for 77% and radionuclide ventriculography (RNV) for 22% of all nuclear cardiology. On a national basis this represents activity levels of 0.86 and 0.25 studies/1,000/year for MPI and RNV, respectively. Of the 171 responding centres, 102 (60%) performed MPI studies and 81 (79%) of these reported that activity was increasing. However, MPI activity was unevenly distributed between hospitals. Two centres accounted for 13% of total MPI; others had far lower activity rates, and 51/102 (50%) centres performed less than 200 MPI studies/year. Comparison with previous surveys showed that nuclear medicine activity had almost doubled since 1990 (it was 6.0 studies/1,000 population in 1990, 9.3 studies/1,000 in 1994 and 11.8 studies/1,000 in 1997). Over the same period, nuclear cardiology activity had also risen, the greatest increase being seen for the last 3 years (it was 0.7 studies/1,000 population in 1990, 0.82 studies/1,000 in 1994 and 1.12 studies/1,000 in 1997). Despite these encouraging figures, MPI activity for 1997 remained well below that recommended by the British Cardiac Society in 1994 (2.6 studies/1,000/year) as adequate to serve the needs of patients with cardiac disease in the UK; it was also below the European average activity for the same year (2.2 studies/1,000/year). The anticipated increased workload for nuclear cardiology is encouraging despite the wide and varied practice of nuclear cardiology around the UK. The nuclear medicine community now needs to address the issues that will prevent it keeping up with demand, such as restricted camera time, excessive waiting lists and outdated equipment, but also to standardise acquisition and reporting techniques so that all studies, wherever performed, will be of a uniformly high standard. PMID- 11914895 TI - Imaging of low-grade bone infection with a technetium-99m labelled monoclonal anti-NCA-90 Fab' fragment in patients with previous joint surgery. AB - Low-grade bone infection represents a serious clinical problem. Diagnostic options are often insufficient, yet the therapeutic implications of proven disease are important, especially in patients with prosthetic joint replacement. Technetium-99m labelled monoclonal anti-NCA-90 granulocyte antibody Fab' fragment (MN3 Fab') has been shown to be useful in bone and joint infection, but there are no data specifically referring to low-grade bone infection. We therefore analysed 38 scans in 30 consecutive patients (age range, 30-85 years; median age, 62 years) referred for suspected low-grade bone infection. There were 17 patients (21 scans) with total hip arthroplasty (THA), six with total knee arthroplasty (TKA), three who had undergone hip or knee surgery for trauma and five (seven scans) with resected hips and no endoprostheses (Girdlestone situations); one of these five patients had been investigated before with THA in situ and another prior to surgery for low-grade coxitis. There were no patients with rheumatoid arthritis as the underlying disease. Results were verified by means of bacteriological cultures, histopathological findings and/or follow-up and compared with the respective Zimmerli scores, which were used for clinical assessment of inflammatory activity. In one patient, the final diagnosis could not be established. One, 5 and 24 h after intravenous injection of up to 1.1 GBq of MN3 Fab', whole-body and planar scans were performed using a dual-head gamma camera. Scans were analysed visually and semiquantitatively adopting an arbitrary score ranging from 0 to 3. There were 13 true positive, 14 true negative and 10 false positive outcomes, yielding an overall sensitivity of 100%, an overall specificity of 58%, an accuracy of 73% and positive and negative predictive values of 57% and 100%, respectively. In patients with THA or TKA, accuracy was 81% and 80%, respectively, while it dropped to 43% in patients with Girdlestone situations owing to a high proportion of false positive findings (4/7) in this subgroup. Scintigraphic score was 1 in all of the false positive and in 11/13 true positive findings. The two remaining true positive findings displayed scintigraphic scores of 2 and 3, respectively. Scintigraphic and Zimmerli scores were loosely correlated (Spearman rho=0.38, P<0.05). Infection was excluded in 22/24 investigations with Zimmerli scores of <6. In this group, there were 13 scintigraphically true negative, nine false positive outcomes, and just two true positive outcomes. In 11/12 investigations with Zimmerli scores of 6 or 7, infection was verified and scintigraphic outcome was accordingly true positive, while the remaining patient was true negative. In conclusion, MN3 Fab' scintigraphy proved to be highly sensitive but not specific in diagnosing low grade infections of the hip and knee regions in patients with previous joint surgery. The method seems reliable in excluding but not in proving the presence of infection. MN3 Fab' scintigraphy should not be applied in patients with Girdlestone situations. Assessment of infection using the Zimmerli score was more reliable than MN3 Fab' scintigraphy in this group of patients without rheumatoid arthritis as the underlying disease. Considering results from the literature concerning leucocyte scintigraphy, MN3 Fab' scintigraphy may be clinically useful in evaluating low-grade bone infection in THA and TKA patients with Zimmerli scores above 5 and concomitant rheumatoid arthritis or other inflammatory diseases. PMID- 11914898 TI - Image registration: an essential tool for nuclear medicine. AB - There is increasing interest in being able to automatically register medical images from either the same or different modalities. Registered images are proving useful in a range of applications, not only providing more correlative information to aid in diagnosis, but also assisting with the planning and monitoring of therapy, both surgery and radiotherapy. The practising nuclear medicine specialist is faced with a dilemma in choosing an appropriate method since the literature in the field is extensive, with conflicting evidence as to what methods are optimal. Although most barriers to implementing registration in routine practice have been removed, there remains a lack of commercial, validated software. The alternative is to install a dual-modality instrument. The objective of this review is to present a general overview of medical image registration with emphasis on the application and issues relevant to nuclear medicine. PMID- 11914899 TI - Metabolic imaging of a solitary pulmonary nodule. PMID- 11914903 TI - Effects of four Chinese herbal extracts on drug-sensitive and multidrug-resistant small-cell lung carcinoma cells. AB - PURPOSE: We examined the pharmacology, cell biology and molecular biology of small-cell lung carcinoma cells treated with four extracts of Chinese herbal medicines. Many cancer patients take these medicines, but their effects at the cellular level are largely unknown. We were especially interested in the effects on drug-resistant cells, as resistance is a significant clinical problem in lung cancer. METHODS: Drug-sensitive (H69), multidrug-resistant (H69VP) and normal lung epithelial cells (BEAS-2) were exposed to extracts from two plants used in Chinese herbal medicine for lung cancer: Glycorrhiza glabra (GLYC) and Olenandria diffusa (OLEN), and to extracts of two commercially available combinations of Chinese herbal medicines, SPES (15 herbs) and PC-SPES (8 herbs). Cytotoxicity was measured in terms of cell growth inhibition (IC(50)). The kinetics of DNA fragmentation after exposure to the herbal extracts was measured by BudR labeling followed by ELISA. Apoptosis was measured by the TUNEL assay followed by flow cytometry. Expression of apoptosis- and cell cycle-related genes was measured by reverse transcription of mRNA followed by filter hybridization to arrays of probes and detection by chemiluminescence. RESULTS: In each case, the four herbal extracts were equally cytotoxic to H69 and H69VP and less cytotoxic to BEAS-2. All four extracts induced DNA fragmentation in the lung carcinoma cells. The kinetics showed DNA fragments released to the medium (an indication of necrosis) in GLYC-exposed cultures, but inside the cells (an indication of apoptosis) in OLEN-, SPES- and PC-SPES-exposed cultures. TUNEL analysis confirmed that exposure to the latter three extracts, but not to GLYC, led to apoptosis. Compared to untreated and GLYC-treated cells, H69 and H69VP cells treated with OLEN, SPES and PC-SPES showed elevated expression of a number of genes involved in the apoptotic cascade, similar to cells treated with etoposide and vincristine. CONCLUSION: The Chinese herbal medicine extracts OLEN, SPES and PC-SPES are cytotoxic to both drug-resistant and drug-sensitive lung cancer cells, show some tumor cell specificity compared to their effect on normal cells, and are proapoptotic as measured by DNA breaks and gene expression. The reaction of the tumor cells to these extracts was similar to their reaction to conventional chemotherapeutic drugs. PMID- 11914904 TI - Dihydropyrimidine dehydrogenase expression in preoperative biopsy and surgically resected specimens of gastric carcinoma. AB - PURPOSE: 5-Fluorouracil (5-FU) is most commonly used to treat patients with gastrointestinal neoplasms, including gastric carcinoma. Dihydroxypyrimidine dehydrogenase (DPD), an enzyme involved in metabolism of 5-FU, is a key factor determining the sensitivity of the tumors to 5-FU. Particularly when preoperative chemotherapy based on 5-FU is attempted, it is critical to know in advance how much DPD the tumor expresses. We investigated preoperative biopsy and surgically resected specimens of gastric carcinomas immunohistochemically for the expression of DPD. METHODS: The study group comprised 55 advanced gastric carcinoma patients who had undergone surgery. Sections of the biopsy and resected specimens were immunostained with anti-DPD polyclonal antibody. DPD immunoreactivity was classified into four groups based on staining intensity expressed as DPD score. We compared the DPD scores of the biopsy specimens with those of the resected specimens. The possible associations between DPD score and survival rate or clinicopathological parameters, including prognostic factors, were also analyzed. RESULTS: The DPD scores of the biopsy specimens correlated with those of the resected specimens (kappa=0.456). In agreement with previous reports, the DPD scores of the gastric carcinomas were not associated with prognosis or with any clinicopathological factor. CONCLUSIONS: It is considered that immunohistochemical analysis of DPD expression in gastric carcinoma using biopsied tissue is a technically feasible method to assess the expression of DPD in the tumor prior to surgical resection. PMID- 11914905 TI - Effects of paclitaxel, cyclophosphamide, ifosfamide, tamoxifen and cyclosporine on the metabolism of methoxymorpholinodoxorubicin in human liver microsomes. AB - The effects of paclitaxel, cyclosporine, cyclophosphamide, ifosfamide and tamoxifen on the metabolism of methoxymorpholinodoxorubicin (MMDx), a novel anticancer agent, were investigated using human liver microsomes. Paclitaxel, tamoxifen and cyclosporine dramatically inhibited MMDx metabolism, whereas ifosfamide had only a slight effect at high concentrations (200-300 microM) and cyclophosphamide had no effect. The inhibition was dependent on the concentrations of both MMDx and the coincubated drug. Thus, with 1 microM MMDx, paclitaxel (5 microM), tamoxifen (1 microM) and cyclosporine (1 microM) decreased the metabolic rate of MMDx by 36%, 53% and 62%, respectively. At higher concentrations (10, 5 and 5 microM, respectively, with paclitaxel, tamoxifen and cyclosporine) the inhibition was 52%, 91% and 91%, respectively. These three drugs preferentially inhibited the formation of three metabolites (M2, M3 and M6) among eight metabolites produced in liver microsomes. The inhibitory concentrations of paclitaxel, tamoxifen and cyclosporine on MMDx metabolism were in the range of those observed in patients upon administration of these drugs, which are known to be CYP3A4 substrates. These findings suggest that CYP3A4 drug substrates and MMDx in combination must be used with caution, particularly in view of the fact that MMDx is considered as a prodrug whose activation is entirely dependent upon metabolic transformation by CYP3A4. PMID- 11914906 TI - XK469, a novel antitumor agent, inhibits signaling by the MEK/MAPK signaling pathway. AB - PURPOSE: XK469 (NSC 697887) is a novel antitumor agent with broad activity against a variety of tumors including drug-resistant tumors. Previous studies have indicated that XK469 is an antiproliferative agent with a low cytotoxic effect in human H116 tumor cells. In this study, we sought to determine the signaling pathways involved in mediating its antiproliferative activity. METHODS: The antiproliferative activity of XK469 was tested using human U-937 leukemia cells in culture. XK469-induced cell cycle arrest was determined using flow cytometric analysis. Phosphorylation/activation of MEK and MAPK was analyzed using immunoblot analyses with specific antibodies against p-MEK and p-MAPK. RESULTS: Cell cycle analysis revealed that XK469 arrested U-937 cells at the G(2)/M phase. Compared with conventional anticancer agents, XK469 showed very low, if any, cytotoxic or proapoptotic effect against U-937 cells. In contrast, treatment of U-937 cells with vinblastine, doxorubicin and m-AMSA resulted in extensive cell death through apoptotic pathways. XK469, but not other agents, potently inhibited the phosphorylation/activation of MEK in U-937 cells cultured in serum-containing medium. XK469 was also able to block the activation of MEK by serum addition in starved U-937 cells. Exposure of cells to XK469 for 1 h was sufficient to inhibit the activation of MEK and its downstream kinase, MAPK. The antiproliferative response to XK469 was correlated with a steady accumulation of cyclins B1 and A, which appeared to be a direct result of G(2)/M arrest. CONCLUSIONS: Our findings suggest that the antiproliferative effect of XK469 is mediated by inhibiting the MEK/MAPK signaling pathways in U-937 human leukemia cells. PMID- 11914907 TI - Population pharmacokinetics of the novel anticancer agent KRN7000. AB - PURPOSE: KRN7000 is a novel anticancer agent, acting through stimulation of the immune system. The first clinical trial with this agent, which included pharmacokinetic studies, has recently been completed. The aim of the study presented here was to develop a population pharmacokinetic model for KRN7000. METHODS: Plasma concentration-time data were gathered from 24 patients enrolled in a phase I trial in which KRN7000 was administered as a weekly slow injection at doses ranging from 50 to 4800 microg/m(2). These data were used to build a pharmacokinetic model using the nonlinear mixed-effect modeling (NONMEM) program. The model was validated by performance of 200 bootstraps. RESULTS: A three compartment model with interindividual variability on the central and two peripheral volumes of distribution (V1, V2 and V3) and on clearance (CL) adequately described the data. The final estimates were: V1 2.34 l, V2 2.61 l, V3 2.13 l, and CL 0.130 l/h. Of 24 covariates tested, including both demographic and pathophysiological factors, none showed a significant relationship with the pharmacokinetic parameters obtained. The bootstrap analysis provided parameter estimates within approximately 15% of the original estimates, indicating stability of the model. CONCLUSION: The pharmacokinetic behavior of KRN7000 in the clinical trial could be described by a three-compartment model. Hence, KRN7000 demonstrates linear pharmacokinetics over the investigated dose range. The pharmacokinetics of KRN7000 are not influenced by patient demographic or pathophysiological characteristics. PMID- 11914908 TI - Alterations in intestinal permeability following the intensified polydrug chemotherapy IFADIC (ifosfamide, Adriamycin, dacarbazine). AB - PURPOSE: The aim of this study was to investigate the severity and time-course of alterations in gastroduodenal and intestinal permeability in relation to nausea/emesis following administration of the highly emetogenic polydrug regimen IFADIC (ifosfamide, Adriamycin, dacarbazine) using a differential lactulose/mannitol absorption (SLM) test. We also assessed the ease of administration and patients' tolerance of the SLM test. METHODS: The SLM test was performed in seven patients with soft tissue sarcomas on days 1, 3 and 14 of cycle I and cycle III of chemotherapy; seven healthy volunteers served as controls. The degree of correlation between the clinical grade of nausea/emesis according to WHO criteria and gastroduodenal permeability, expressed in terms of urinary sucrose excretion, and intestinal permeability, expressed in terms of the permeability index (urinary lactulose to mannitol permeability ratio), was also assessed. RESULTS: The permeability index values were significantly different (P < or =0.01) on days 1, 3 and 14 during both cycles of chemotherapy. The median permeability index on day 3 was higher (P < =0.01) in patients with nausea/emesis than in those without symptoms. Additionally, the permeability index when nausea was present (day 3) was higher (P < or =0.01) than when nausea/emesis was absent (days 1 and 14). In 59% of patients the increased permeability index on day 3 was accompanied by nausea/emesis of WHO grade 3. Gastroduodenal permeability did not alter consistently following chemotherapy. CONCLUSIONS: Our study confirms an acute, transient increase in intestinal permeability following the polydrug regimen IFADIC, accompanied by nausea/emesis of WHO grade 3 in the majority of patients. Normal intestinal permeability was achieved on day 14 in all patients, thus allowing intensified 2-weekly treatment administration. The SLM test may be recommended as a feasible test for the objective assessment of alterations in intestinal permeability following chemotherapy administration. PMID- 11914909 TI - Lack of activity of stealth liposomal doxorubicin in the treatment of patients with anthracycline-resistant breast cancer. AB - PURPOSE: We conducted a single-institution phase II clinical trial to determine the objective response rate, duration of response, time to progression, and overall survival in patients with anthracycline-resistant breast cancer treated with Doxil. PATIENTS AND METHODS: Patients with metastatic breast cancer were eligible if they had disease progression while receiving an anthracycline containing regimen or developed evidence of metastatic disease during or within 6 months after the last cycle of an anthracycline-containing adjuvant regimen. Prior treatment with liposomal doxorubicin was not allowed. Patients received a dose of 50 mg/m(2) infused every 4 weeks via a peripheral vein or central line. Doxil was administered for a total of six cycles or until disease progression. RESULTS: We treated 11 patients with stage IV breast cancer of whom two had never received chemotherapy for their metastatic disease. Most had a performance status of 1 and had visceral involvement as their dominant site of disease. All patients had received prior therapy with doxorubicin. No clinical evidence of congestive heart failure or cardiac toxicity was observed. The most common toxicities were nonhematologic and were mostly grade 1/2. These included fatigue, nausea, vomiting, and stomatitis. Significant myelosuppression was only observed in one patient. No complete or partial response was observed. There were two patients who had a minimal response and two other patients who had evidence of stable disease. CONCLUSION: Doxil was well tolerated with minimal toxicity. However, the lack of antitumor activity in anthracycline-resistant breast cancer patients indicates that further evaluation in this patient population is not warranted. PMID- 11914910 TI - Low dose--high dose: what is the right dose? Pharmacokinetic modeling of etoposide. AB - PURPOSE: Some clinical studies on etoposide (Eto) have shown marked schedule dependency of the effect (starting at about 1 mg/l) and toxicity (over about 10 mg/l) whereas other studies have not confirmed these results. What are the conclusions we can draw from these inconsistent results when developing new low dose (LD) and high-dose (HD) Eto schedules? METHODS: A pharmacokinetic model for Eto based on individual pharmacokinetic data was used to simulate different LD (450 mg/m(2)) and HD (1800 mg/m(2)) schedules. The duration of exposure and the AUC of relevant concentration ranges (>1 mg/l, 1-10 mg/l, >10 mg/l) as well as peak levels were calculated in relation to the standard low dose (150 mg/m(2) over 2 h daily for 3 days). RESULTS: The fourfold dose increase from the LD to the HD schedule was associated with a complementary increase in total AUC. However, variations in infusion time for the HD schedule were associated with large differences in AUC distribution and drug exposure with constant total AUC. Short infusions (0.5 h and 4 h) resulted in extreme peak levels (factors of 17.6+/-1.5 and 8.7+/-0.4 compared to the standard LD schedule) and an AUC >10 mg/l (factors of 17.5+/-4.9 and 17.2+/-4.8, and 83+/-2.4% and 82+/-2.4% of the corresponding total AUC), and the 96-h infusion yielded a long duration of exposure to concentrations >10 mg/l (factor 7.9+/-2.6), whereas continuous i.v. infusion over 55+/-11 days was associated with a multiple increase in the duration of exposure to "standard" drug concentrations (1 mg/l ). CONCLUSIONS: According to evidence against schedule dependency, only target dose and pharmacokinetic variability would be appropriate rationales for Eto dosing. However, arguing for schedule dependency, simulated pharmacokinetic profiles for new Eto schedules over a wide range support that Eto-containing regimens should be designed on the basis of clear pharmacokinetic hypotheses of target levels, exposure times and AUC distributions, to allow subsequent development of pharmacokinetic/pharmacodynamic modeling. PMID- 11914911 TI - Cellular levels of aldehyde dehydrogenases (ALDH1A1 and ALDH3A1) as predictors of therapeutic responses to cyclophosphamide-based chemotherapy of breast cancer: a retrospective study. Rational individualization of oxazaphosphorine-based cancer chemotherapeutic regimens. AB - PURPOSE: In preclinical models, established molecular determinants of cellular sensitivity to cyclophosphamide, long a mainstay of chemotherapeutic regimens used to treat breast cancers, include the aldehyde dehydrogenases that catalyze the detoxification of this agent, namely, ALDH1A1 and ALDH3A1. As judged by bulk quantification of relevant catalytic activities, as well as of relevant proteins (ELISAs), tissue levels of these enzymes vary widely in primary and metastatic breast malignancies. Thus, interindividual variation in the activity of either of these enzymes in breast cancers could contribute to the wide variation in clinical responses obtained when such regimens are used to treat these malignancies. Direct evidence for this notion was sought in the present investigation. METHODS: Cellular levels of ALDH1A1 and ALDH3A1 in 171 repository human breast tumor (122 primary and 49 metastatic) samples were semiquantified using immunocytochemical staining. Clinical responses were retrieved from the archived medical records of each of 48 metastatic breast cancer sample donors, 26 of whom had been treated with a cyclophosphamide-based chemotherapeutic regimen subsequent to tumor sampling and 22 of whom had not. The premise that cellular levels of ALDH1A1 and/or ALDH3A1 predict clinical responses to cyclophosphamide based chemotherapeutic regimens was submitted to statistical analysis. RESULTS: Confirming an earlier report, ALDH1A1 and ALDH3A1 levels varied widely in both primary and metastatic breast tumor cells. When measurably present, each of the enzymes appeared to be evenly distributed throughout a given tumor cell population. Retrospective analysis indicated that cellular levels of ALDH1A1, but not those of ALDH3A1, were (1) significantly higher in metastatic tumor cells that had survived exposure to cyclophosphamide than in those that had not been exposed to this drug, and (2) significantly higher in metastatic tumors that did not respond (tumor size did not decrease or even increased) to subsequent treatment with cyclophosphamide-based chemotherapeutic regimens than in those that did respond (tumor size decreased) to such regimens. The therapeutic outcome of cyclophosphamide-based chemotherapy corresponded to cellular ALDH1A1 levels in 77% of cases. The frequencies of false-positives (cyclophosphamide-based chemotherapy not effective when a low level of ALDH1A1 predicted it would be) and false-negatives (cyclophosphamide-based chemotherapy effective when a high level of ALDH1A1 predicted it would not be) were 0.00 and 0.43, respectively. Thus, partial or complete responses to cyclophosphamide-based chemotherapy occurred 2.3 times more often when the ALDH1A1 level was low than when it was high. CONCLUSIONS: Given (1) the wide range of ALDH1A1 levels observed in malignant breast tissues, (2) that ALDH1A1 levels in primary breast tumor tissue, as well as those in normal breast tissue, directly reflect ALDH1A1 levels in metastatic breast tumor cells derived therefrom, and (3) the findings reported here, measurement of ALDH1A1 levels in primary breast malignancies and/or normal breast tissue prior to the initiation of chemotherapy is likely to be of value in predicting the therapeutic potential, or lack of potential, of cyclophosphamide and other oxazaphosphorines, e.g. ifosfamide, in the treatment of primary, as well as metastatic, breast cancer, thus providing a rational basis for the design of individualized therapeutic regimens for this disease. Failure to observe the expected inverse relationship between clinical responses to cyclophosphamide based chemotherapeutic regimens and ALDH3A1 levels was probably because even the highest breast tumor tissue ALDH3A1 level thus far reported appears to be below the threshold level at which ALDH3A1-catalyzed detoxification of oxazaphosphorines becomes pharmacologically meaningful. However, ALDH3A1 levels in certain other malignancies, e.g. those of the alimentary tract and lung, may be of a sufficient magnitude in that regard. PMID- 11914912 TI - P-glycoprotein inhibition by the multidrug resistance-reversing agent MS-209 enhances bioavailability and antitumor efficacy of orally administered paclitaxel. AB - PURPOSE: Recent studies in humans and mice have demonstrated that intestinal P glycoprotein plays a causative role in the limited absorption of orally administered paclitaxel. Multidrug resistance (MDR)-reversing agents, such as cyclosporin A and PSC 833, are known to increase the systemic exposure to orally administered paclitaxel by enhancing absorption in the intestinal tract and decreasing elimination via the biliary tract. In this study, we demonstrated that coadministration of the MDR-reversing agent MS-209, which is known to inhibit P glycoprotein function by direct interaction, improved the bioavailability of orally administered paclitaxel and consequently enhanced its antitumor activity. METHODS: The pharmacokinetics of paclitaxel were examined by measuring [(3)H]paclitaxel in plasma drawn from rats and mice given the drug with or without MS-209. The influence of MS-209 on the intestinal transport of [(3)H]paclitaxel was studied using a human colorectal cancer cell line, Caco-2. The in vivo efficacy of orally administered paclitaxel in combination with MS-209 was further evaluated in B16 melanoma-bearing mice. RESULTS: The plasma concentration of [(3)H]paclitaxel following oral administration was significantly increased by coadministration of MS-209 at 100 mg/kg in both rats and mice. In rats, the AUC of [(3)H]paclitaxel following oral administration was strikingly increased (1.9-fold) by coadministration of MS-209, whereas the AUC of [(3)H]paclitaxel following i.v. injection was slightly increased (1.3-fold) by MS 209. The increase in apparent bioavailability of oral paclitaxel due to MS-209 was 1.4-fold. To demonstrate this enhancing action in vitro, we studied the influence of MS-209 on the transport of [(3)H]paclitaxel using Caco-2 cells, which is a well-known model of intestinal efflux. The transport of [(3)H]paclitaxel across the Caco-2 monolayer was markedly inhibited in the presence of MS-209, and the apparent K(i)of MS-209 for the active transport of [(3)H]paclitaxel was 0.4 microM. Moreover, paclitaxel administered orally at 100 mg/kg per day with MS-209 at 100 mg/kg per day showed significant antitumor activity in B16 melanoma-bearing mice, whereas paclitaxel administered orally alone at the same dose showed no antitumor activity. These results suggest that the coadministration of MS-209 improved low systemic exposure to paclitaxel through inhibition of P-glycoprotein, which is involved in drug excretion via the intestinal tract, resulting in a clear antitumor activity of paclitaxel administered orally. CONCLUSION: The present study suggests that coadministration of MS-209 may be a useful way to improve the bioavailability of drugs not suitable for oral administration due to elimination via the intestinal tract. PMID- 11914913 TI - Determinants of the cytotoxicity of irinotecan in two human colorectal tumor cell lines. AB - PURPOSE: Irinotecan is a drug of the camptothecin family that has proven activity in advanced colon cancer, with about 20% responses in untreated as well as in 5 fluorouracil-resistant tumors. Irinotecan is considered as a prodrug which needs to be activated to SN-38 by carboxylesterases to become able to interact with its target, topoisomerase I. The work reported here intended to identify the determinants of the cytotoxicity of irinotecan in two human colorectal tumor cell lines, LoVo and HT-29, at the level of the target of the drug and at the level of the availability of the active metabolite to the target. RESULTS: The cytotoxicity of irinotecan and SN-38 markedly differed in the two cell lines: irinotecan IC(50) values were 15.8 microM for LoVo cells and 5.17 microM for HT 29 cells; SN-38 IC(50) values were 8.25 n M for LoVo cells and 4.50 n M for HT-29 cells. Topoisomerase I expression (at the mRNA and the protein levels) and catalytic activity were similar in the two cell lines. Irinotecan induced similar amounts of cleavable complexes at its IC(50) in both cell lines. SN-38 induced a concentration-dependent formation of cleavable complexes, which was not significantly different in the two cell lines. Expression of the carboxylesterase CES1 was higher in HT-29 than in LoVo cells. Expression of the carboxylesterase gene CES2 was comparable in the two cell lines and much higher than CES1 gene expression. Carboxylesterase activity was extremely low using p nitrophenylacetate as a substrate (1.45 and 1.84 pmol/min per mg proteins) and could not even be detected using irinotecan as a substrate. Cell accumulation of irinotecan was markedly different, reaching consistently higher levels in HT-29 cells than in LoVo cells. CONCLUSIONS: Our results indicate that (1) the cytotoxicity of irinotecan was likely due to the drug itself and not to its metabolite SN-38, and (2) that irinotecan uptake was more predictive of its cytotoxicity than topoisomerase I availability and activity in these two cell lines. PMID- 11914914 TI - Biliary transport of irinotecan and metabolites in normal and P-glycoprotein deficient mice. AB - PURPOSE: The extensive and unpredictable biliary excretion of CPT-11 and its metabolites, SN-38 and SN-38 glucuronide (SN-38G) may contribute to the wide interpatient variability reported in the disposition and gastrointestinal toxicity of CPT-11. We studied the role of P-glycoprotein (P-gp) in in vivo biliary excretion of CPT-11, SN-38 and SN-38G in mice lacking mdr1-type P-gp [ mdr1a/1b(-/-)] in the presence of the multidrug resistance (MDR) reversal agent, PSC833. METHODS: Wild-type (Wt) and mdr1a/1b(-/-) mice ( n=3 or 4) were treated intragastrically with PSC833 (50 mg/kg) or vehicle 2 h prior to i.v. CPT-11 dosing (10 mg/kg), and bile samples were collected. RESULTS AND CONCLUSIONS: P-gp was found to play an important role in CPT-11 biliary excretion, as there was a significant (40%, P<0.05) decrease in its biliary recovery in 90 min in mdr1a/1b( /-) mice (6.6+/-0.6% dose) compared with Wt mice (11+/-1.2%). This also implied a major role of other undetermined non-P-gp-mediated mechanism(s) for hepatic transport of CPT-11, which was inhibited by PSC833 (1.8+/-0.8% with PSC833, 6.6+/ 0.6% without PSC833) in mdr1a/1b(-/-) mice. SN-38 and SN-38G biliary transport was unchanged in mice lacking P-gp after vehicle treatment, indicating a lack of P-gp mediation in their transport. PSC833 significantly reduced (56-89%) SN-38 and SN-38G biliary transport in Wt and mdr1a/1b(-/-) mice, suggesting that PSC833 may be a candidate to modulate biliary excretion of SN-38 with potential use in reducing CPT-11 toxicity. PMID- 11914915 TI - A phase II trial of methotrexate-human serum albumin (MTX-HSA) in patients with metastatic renal cell carcinoma who progressed under immunotherapy. AB - INTRODUCTION: Renal cell carcinoma (RCC) has a poor prognosis when metastasized to distant sites, although immunotherapy may offer a prolongation of survival in selected patient groups. Unfortunately, no treatment options remain when immunotherapy fails. In this phase IIa trial the tolerability and efficacy of the antifolate drug methotrexate-human serum albumin (MTX-HSA) were evaluated in patients with metastatic RCC who progressed after first-line immunotherapy. PATIENTS AND METHODS: A total of 17 patients started treatment, and 14 (12 men, 2 women) were evaluable for response according to the phase IIa Gehan design. Patients had had prior tumor nephrectomy, were in relatively good general condition, had no impairment of renal, liver or bone marrow function, and had progressive metastatic disease after treatment with interferon-alpha (IFN-alpha) with or without cis-retinoic acid (EORTC protocols 30951 and 30947). MTX-HSA was given once a week intravenously on an outpatient basis at a dose of 50 mg/m(2). The treatment interval was prolonged in those patients who had not yet recovered from previous toxicities. RESULTS: Toxicity was manageable, relatively mild to moderate and reversible in most cases. Grade 2/3 mucositis (10/17) and grade 3 elevated transaminase levels (4/17) were most frequent, and in only one patient was a grade 4 thrombocytopenia reported. Of three inevaluable patients, one discontinued treatment due to drug-related toxicities. The mean administration interval was 12.1 days, and 7 of 14 evaluable patients had treatment intervals of 1 or 2 weeks. No objective responses were seen, although eight patients had stable disease (stabilization >2 months) for up to 8 months (median 121 days). CONCLUSION: MTX-HSA was generally well tolerated and can be given on an outpatient basis, but no objective responses were seen in patients with metastatic RCC who had progressed after previous immunotherapy. PMID- 11914916 TI - Ultrasensitive pharmacological characterisation of the voltage-gated potassium channel K(V)1.3 studied by single-molecule fluorescence microscopy. AB - The determination of pharmacologically relevant constants is crucial in order to understand the effects of compounds interacting with various membrane receptors. In this report we study a venom component of the Central American scorpion Centruroides limbatus, a short peptide termed hongotoxin(1) (HgTX(1)), which specifically binds to the voltage-gated potassium channel K(V)1.3 at a molecular stoichiometry of 1:1. A toxin analogue (HgTX(1)-A19C) was subjected to fluorescence labelling studies with Cy5. Utilising an ultrasensitive microscopic method (single-dye tracing; SDT) we were able to directly visualise HgTX(1)-A19C Cy5 binding to the voltage-gated potassium channel K(V)1.3 on Jurkat cells at the single molecule level. For the first time, this approach allowed the determination of both the dissociation constant (K(D)) and the off-rate (k(off)) of HgTX(1)-A19C-Cy5 on living cells. In order to validate this novel approach, the data obtained with SDT were correlated to radioligand binding studies performed under identical conditions using a radioiodinated HgTX(1) analogue. PMID- 11914917 TI - Expression and induction of the stress protein alpha-B-crystallin in vascular endothelial cells. AB - Stress-induced development of enhanced tolerance against various kinds of stresses has been observed in vascular endothelial cells as well as in several other cell types. Stress proteins are thought to play a key role in the development of stress tolerance. In this study we show that endothelial cells of various sources contain the major stress protein of the eye lens, alphaB crystallin. In the mouse myocardial microvascular cell line, MyEnd, alphaB crystallin as well as the heat shock proteins HSP 70i and HSP 25 display a low constitutive expression but can be significantly upregulated by sodium arsenite stress. Osmotic stress also resulted in strong upregulation of alphaB-crystallin and HSP 70i but not of HSP 25. Both osmotic and arsenite stress resulted in significant stress tolerance of MyEnd cells against glucose deprivation as assayed by lactate dehydrogenase release and overall cellular morphology. Development of stress tolerance without induction of HSP 25 indicates that HSP 25 is not essential for the protective effect. MyEnd cells from alphaB-crystallin-/- mice displayed a similar degree of stress tolerance showing that alphaB crystallin is dispensable for protection of cells against energy depletion. The functional role of alphaB-crystallin in endothelial cells needs to be further elucidated. In our experiments HSP 70i turned out to be the only potential candidate of the stress proteins assayed to be involved in the development of tolerance against energy depletion. PMID- 11914918 TI - Immunohistochemical localization of acyl-CoA hydrolase/thioesterase multigene family members to rat epithelia. AB - Acyl-CoA hydrolases cleave acyl-CoA thioesters to free fatty acids and coenzyme A. The potency of these enzymes may serve to modulate cellular levels of acyl CoAs to affect various cellular functions, including lipid metabolism. In this study, we investigated the tissue distribution of this multigene family of enzymes, focusing on cytosolic (CTE-I) and mitochondrial acyl-CoA thioesterases (MTE-I) in adult rats, using an anti-CTE-I antibody which recognizes both the isoforms. Western blotting detected them mainly in organs closely related to fatty acid oxidation, of which kidney contained the highest levels of both enzymes. Immunohistochemistry localized the enzymes primarily in the proximal tubules, where a large energy demand is expected and fatty acids represent a major fuel, correlating well with the intrarenal distribution of peroxisomal beta oxidation. In situ hybridization suggested colocalization of CTE-I and MTE-I in the kidney. The immunoreactivity was also found in various epithelial tissues in the body, including Harderian gland and sebaceous gland. These results demonstrated the distribution of CTE-I and MTE-I in a wide variety of rat tissues, primarily characterized by an epithelial localization, being consistent with their involvement in fatty acid metabolism. PMID- 11914920 TI - Protamine-1 and -2 mRNA in round spermatids is associated with RNA-binding proteins. AB - RNA-binding proteins in round spermatids have previously been assigned to the coding sequence of Prm1- and Prm2-mRNA. To further characterize this protein-RNA interaction, prior to cDNA synthesis, microdissected cell profiles were digested with different proteases exhibiting a specific cleavage site followed by both conventional and real-time quantitative PCR. Best results were obtained with proteinase K and A followed by factor Xa protease, genenase I, and proteases V8. While enterokinase revealed PCR signals solely for Prm2, no amplification signal was obtained using chymotrypsin. These data suggest a protein segment rich in basic amino acids to be important for the binding to Prm1- and Prm2-mRNA. The fact that phenanthroline treatment instead of protease digestion also resulted in amplification signals suggests the involvement of zinc-finger-like protein-RNA interactions. Employing different primer pairs, RNA-binding proteins were shown to be localized at the 5' end of Prm1- and Prm2-mRNA. Since protein-RNA interactions are a common principle of posttranscriptional regulation of gene expression, the combination of microdissection, protease digestion, and real-time quantitative PCR provides a suitable tool for its investigation in a cell type specific manner. Furthermore, the presence of RNA-binding proteins within the coding sequence of mRNAs demands proteinase K treatment prior to cDNA synthesis, a compelling necessity for the study of gene expression. PMID- 11914919 TI - Expression of the Thomsen-Friedenreich antigen and of its putative carrier protein mucin 1 in the human placenta and in trophoblast cells in vitro. AB - The Thomsen-Friedenreich (TF) antigen (or, more precisely, epitope Galbeta1 3GalNAcalpha-O-) has been known for a long time as a carcinoma-associated antigen. In normal tissues the occurrence of TF antigen is restricted to a few immunologically privileged areas. Here we report on the identification of the TF epitope and its putative carrier protein mucin 1 (MUC1) in human placental tissue, on isolated trophoblast cells in vitro and on trophoblast tumour cell lines BeWo and Jeg3. Cryosections of placental and decidual tissues of the first, second and third trimester were double stained with monoclonal antibodies directed against the TF epitope (IgM) and against MUC1 (IgG). In the first trimester of pregnancy we found strong expression of TF antigen and MUC1 at the apical side of the syncytiotrophoblast directed towards the maternal blood. This expression was consistent in the second trimester of pregnancy, and to a lesser degree in the third trimester. In addition, we found positive staining for TF antigen and MUC1 on extravillous trophoblast cells in the decidua during the first and second trimester of pregnancy. Trophoblast tumour cells of the cell line BeWo, which form a syncytium in vitro, were also positive for TF antigen and MUC1, whereas Jeg3 cells, which are unable to form a syncytium, expressed only MUC1. Freshly isolated trophoblast cells from first trimester placentas showed strong staining for MUC1; however, only a few of these cells (less than 1%) were positive for TF antigen, and might consist of digested fragments of the syncytium. In summary, TF antigen and MUC1 are expressed by the syncytiotrophoblast at the feto-maternal interface and by extravillous trophoblast cells invading the decidua, whereas villous cytotrophoblast cells in situ as well as freshly isolated trophoblast cells from first trimester placentas only express MUC1 but not TF antigen. PMID- 11914921 TI - Tissue localization of TGFalpha and apoptosis are inversely related in colorectal tumors. AB - Expression of TGFalpha and the EGF receptor was studied in relation to apoptosis in human colorectal mucosa and premalignant and malignant tumors. In normal mucosa the proteins colocalized both in the proliferation compartment and at the luminal pole of the crypts in cells committed to undergo apoptosis. While staining for the EGF receptor was increased in premalignant and malignant lesions, TGFalpha was undetectable in aberrant crypt foci as well as large areas of adenomas. Incidence of apoptosis (AI) was high in these areas ranging from 8.83-24.59. Adenomas did, however, contain islands of high TGFalpha expression where AI was decreased to a range of 0.76-4.00 (decreased at P=0.0027). In carcinomas TGFalpha expression was increased above both normal and adenoma levels corresponding to the decrease in apoptosis in the malignant tumors. Tissue localization of TGFalpha and AI were still inversely related ( P=0.022), but interpatient variability was much larger than for adenomas. The data indicate that TGFalpha is the main survival factor in premalignant tumor cells of the colon, while additional factors moderate its effect in carcinomas. This suggests the possibility of targeting the EGF receptor pathway not only for treatment but also for the reversal of adenoma growth and the prevention of malignant colorectal tumors. PMID- 11914923 TI - Morphological phenotyping of enteric neurons using neurofilament immunohistochemistry renders chemical phenotyping more precise in porcine ileum. AB - The aim of the study was: (1) to test the suitability of neurofilament (NF) immunohistochemistry for representing the shapes of morphologically defined neuron types in the pig ileum myenteric plexus, (2) to estimate the proportions of these neuron types as related to the whole myenteric neuron population and (3) to demonstrate the usefulness of a refined morphological classification of enteric neurons on the paradigm of calcitonin gene-related peptide (CGRP) immunoreactive neurons. So far, immunoreactivity for this peptide was supposed to be present in the pig enteric nervous system only in type II neurons. Ileal whole mounts of two pigs were stained with the cuprolinic blue (CB) method and, thereafter, incubated with an antibody pool against NF proteins (70, 160 and 210 kDa), visualised with a fluorochrome-tagged secondary antibody. The structural representation of morphologically defined myenteric neuron types typical for pig ileum (Stach I, II, IV, V and VI) was equivalent to their silver impregnated image, as demonstrated in previous studies. Counts of CB-stained neurons revealed between 2,526 and 2,662 neurons per square centimetre in one pig and between 2,027 and 2,763 in the other. As related to these total neuron numbers, the proportions of type I neurons were 1.7% and 1.5%, of type II neurons 7.2% and 7.9%, of type IV neurons 1.9% and 2.4%, of type V neurons 1.1% and 1.5%, and of type VI neurons 1.3% each. These values are generally comparable with those estimated earlier on silver impregnated material. Double labelling for NF and CGRP indicated that CGRP-immunoreactive smooth contoured neurons with long processes could be subdivided into two distinct morphological neuron types, i.e. type II and type V. We conclude that NF immunohistochemistry is an appropriate tool for representation of morphologically defined enteric neuron types in the pig. Combination of this technique with immunohistochemistry for neuroactive substances may be useful for making both morphological and chemical classification schemes mutually more precise. PMID- 11914922 TI - Nuclear envelope-limited chromatin sheets are part of mitotic death. AB - Nuclear envelope-limited chromatin sheets (ELCS) are enigmatic membranous structures of uncertain function. This study describes the induction of ELCS in p53 mutated Burkitt's lymphoma cell lines after treatment with irradiation or the microtubule inhibitor, SK&F 96365. Both treatments evoked similar mitotic death, involving metaphase arrest followed by extensive endopolyploidisation and delayed apoptosis, although the kinetics were different. We found that induction of ELCS and nuclear segmentation correlated with the amount and kinetics of M-phase arrest, mitosis restitution and delayed apoptosis of endopolyploid cells. In metaphases undergoing restitution, ELCS are seen participating in the restoration of the nuclear envelope, mediating the attachment of peripheral chromatin to it. In interphase cells, ELCS join nuclear segments, ectopically linking and fusing with heterochromatin regions. In cells with segmented nuclei, continued DNA replication was observed, along with activation and redistribution of Ku70, suggestive of non-homologous DNA end-joining. Induction of ELCS also parallels the induction of cytoplasmic stacked membrane structures, such as confronting cisternae and annulate lamellae, which participate in the turnover and degeneration of ELCS. The results suggest that arrest at a spindle checkpoint and the uncoupling of mitosis from DNA replication lead to the emergence of ELCS in the resulting endopolyploid cells. PMID- 11914924 TI - Modulation of hyaluronan in the migratory pathway of mouse primordial germ cells. AB - In the present report we followed the distribution of hyaluronan during the phases of separation, migration, and colonization of the primordial germ cell migratory process. Hyaluronan was detected by the use of two cytochemical methods: (1) ruthenium hexammine trichloride (RHT) associated with enzymatic treatment with hyaluronate lyase and (2) a binding specific probe for hyaluronan. After RHT treatment the proteoglycans and/or glycosaminoglycans were observed as a meshwork formed by electron-dense granules connected by thin filaments. After enzymatic digestion, no filaments could be detected in the migratory pathway. Quantitative analysis showed a close correlation between cell migration and the concentration of RHT-positive filaments. It was also shown that high amounts of hyaluronan were expressed in the separation phase and migration phases whereas during the colonization phase the amount of hyaluronan was clearly diminished. This study showed that the presence of primordial germ cells in each compartment of the migratory pathway was always accompanied by a high expression of hyaluronan. These results indicate that hyaluronan is an important molecule in the migratory process, providing the primordial germ cells with a hydrated environment that facilitates their movement toward the genital ridges. PMID- 11914925 TI - Interactions between Zn and Cu in LEC rats, an animal model of Wilson's disease. AB - The effect of oral Zn treatment was studied in the liver and kidneys of 26 male Long-Evans Cinnamon (LEC) rats (mutant animals, 5 weeks old) in relation to both the interaction between Zn and Cu and the localisation and concentration of metallothionein (MT). Rats receiving 80 mg zinc acetate daily by gavage and control rats receiving no treatment were killed after 1 or 2 weeks. By immunohistochemical and analytical chemical techniques we revealed that treated rats had higher levels of MT in the hepatic and renal cells compared to untreated ones. Tissue Zn concentrations were significantly higher in treated rats compared to untreated whereas Cu concentrations decreased in the liver and kidneys as indicated by analytical chemical analyses. MT levels also decreased with treatment period. A histochemical procedure, obtained using autofluorescence of Cu-metallothioneins, confirms these findings: after 2 weeks, the signal decreased in both the liver and kidney sections. This gives a greater understanding of the mechanism of Cu metabolism in the two tissues considered. These results suggest that Zn acts both to compete for absorption on the luminal side of the intestinal epithelium and to induce the synthesis of MT. PMID- 11914928 TI - The MEN 1 syndrome: the actual role of genetic testing on the timing and extent of surgery. PMID- 11914926 TI - RANK ligand, RANK, and OPG expression in type II collagen-induced arthritis mouse. AB - Rheumatoid arthritis (RA) is a systemic disorder characterized by synovial inflammation and subsequent destruction and deformity of synovial joints. The articular lesions start with synovitis, focal erosion of unmineralized cartilage, and then culminate in the destruction of subarticular bone by pannus tissue. Periarticular osteopenia and systemic osteoporosis follow as late complications of RA. Osteoclasts, specialized cells that resorb bone, play a central role in developing these osteolytic lesions. To elucidate the mechanism of osteoclastogenesis and bone destruction in autoimmune arthritis, we investigated the expression of RANK ligand (RANKL), RANK, and osteoprotegerin (OPG) mRNA in a mouse type II collagen-induced arthritis (CIA) model by in situ hybridization. The results indicated that most of the TRAP-positive mono- and multinucleated cells in the inflamed and proliferating synovium and in the pannus were RANK positive authentic osteoclasts and their precursors. In the inflamed synovium and pannus of the mouse CIA model, synovial fibroblastic cells around these RANK positive cells were strongly positive for RANKL. Moreover, RANKL-positive osteoblasts on the endosteal bone surface, at a distance from the affected synovial joints, increased significantly in the mouse CIA model prior to periarticular osteopenia and systemic osteoporosis. These data indicated that the RANKL-RANK system plays an important role for osteoclastogenesis in both local and systemic osteolytic lesions in autoimmune arthritis, and can therefore be a good target for therapeutic intervention. PMID- 11914929 TI - Clinical and molecular diagnosis of multiple endocrine neoplasia type 1. AB - Multiple endocrine neoplasia type 1 (MEN1) is a classic hereditary tumor syndrome characterized by a genetic predisposition to develop a variety of neuroendocrine neoplasias and hormone excess syndromes. The disease is caused by inactivating mutations of the MEN1 tumor suppressor gene, detectable in >95% of MEN1 families. The distinction of MEN1-associated tumors from sporadic neuroendocrine neoplasias is clinically important for providing optimal surgical and medical therapy, appropriate clinical follow-up, and counsel for affected patients and their families. Since MEN1 gene analysis became available in 1997, new diagnostic approaches have evolved in clinical management, to be reviewed in this article. Genetic screening of MEN1 families will allow definition of individual disease risk at a preclinical stage, thus helping to allocate medical resources and treatment as individually needed. These new diagnostic approaches are expected to reduce MEN1-associated morbidity and mortality, health care expenses, and psychological disease burden. PMID- 11914930 TI - Primary hyperparathyroidism in MEN 1--how radical should surgery be? AB - Primary hyperparathyroidism is the most common manifestation of MEN 1 syndrome. The management of these patients is complex due to the underlying disease process, which predisposes patients to persistent and recurrent disease. The surgical treatment of patients with MEN 1 and hyperparathyroidism can therefore be considered to be palliative in nature. The basic principles of surgery include (1) obtaining and maintaining normocalcaemia for the longest time possible, avoiding persistent/recurrent hypercalcaemia, (2) avoiding surgically induced hypocalcaemia, and (3) facilitating future surgery for recurrent disease. Two approaches have been described as the best practice for patients with hyperparathyroidism in MEN 1: subtotal parathyroidectomy, leaving a remnant of no more than 60 mg in the neck, and total parathyroidectomy with immediate autotransplantation of 10-20 1 mm(3) pieces of parathyroid tissue. Both approaches should be combined with efforts to exclude supernumerary glands and ectopic parathyroid tissue by including resection of fatty tissue from the central neck compartment and thymectomy in all patients. Cryopreservation of parathyroid tissue should be performed whenever facilities are available. In patients with persistent or recurrent disease, an attempt to obtain total elimination of cervical parathyroid tissue is justified, combined with cryopreservation of parathyroid tissue. As radical as surgery is for hyperparathyroidism in MEN 1, the surgeon must take steps to avoid permanent hypoparathyroidism, which in young patients may be worse than the disease itself. PMID- 11914931 TI - Timing and extent of surgery in symptomatic and asymptomatic neuroendocrine tumors of the pancreas in MEN 1. AB - Pancreaticoduodenal tumors develop in a majority of patients with multiple endocrine neoplasia type 1 (MEN 1) and have a pronounced effect on life expectancy as the principal cause of disease related death. Previous discussion of therapy has focused mainly on syndromes of hormone excess and especially the management of MEN 1 associated Zollinger-Ellison syndrome (ZES). The syndromes of hormone excess, however, may be late features of the endocrinopathy and, when developed, indicate presence of metastases in more than one-third of patients. Recent possibilities for genetic diagnosis have emphasized requirements of prophylactic operation for prevention of malignant development. We recommend screening with biochemical markers and endoscopic ultrasound for early detection, and strong efforts of operative tumor removal before metastases have occurred. Surgery is generally recommended in patients with or without hormonal syndromes in the absence of spread hepatic metastases. Operative procedures include enucleation of tumors in the head of the pancreas, excision of duodenal gastrinomas together with clearance of lymph gland metastases, and as prophylaxis against tumor recurrence combination with distal 80% subtotal pancreatic resection. More extensive surgical tumor reduction is believed to reduce the risks for malignant progression of the pancreaticoduodenal tumors, but this requires further evaluation in MEN 1. PMID- 11914932 TI - NSC-631570 (Ukrain) in the palliative treatment of pancreatic cancer. Results of a phase II trial. AB - BACKGROUND: NSC-631570 (Ukrain) is a semisynthetic compound of thiophosphoric acid and the alkaloid chelidonine from the plant Chelidonium majus. It has been used in complementary herbal medicine for more than 20 years for the treatment of benign and malignant tumors. PATIENTS/METHODS: Between August 1999 and June 2001, 90 patients with histologically proven unresectable pancreatic cancer were randomized in a monocentric, controlled, randomized study. Patients in arm A received 1000 mg gemcitabine/m2, those in arm B received 20 mg NSC-631570, and those in arm C received 1000 mg gemcitabine/m2 followed by 20 mg NSC-631570 weekly. End point of the study was overall survival. RESULTS: In all three arms therapy was well tolerated and toxicity was moderate. At the first re-evaluation in arm A 32%, in arm B 75%, and in arm C 82% showed no change or partial remission according to WHO criteria (arm A versus arm B: P<0.01, arm A versus arm C: P<0.001). Median survival according to Kaplan-Meier analysis was in arm A 5.2 months, in arm B 7.9 months, and in arm C 10.4 months (arm A versus arm B: P<0.01, arm A versus arm C: P<0.01). Actuarial survival rates after 6 months were 26%, 65% and 74% in arms A B and C, respectively (arm A versus arm B: P<0.05, arm A versus arm C P<0.01). CONCLUSION: We could show that in unresectable advanced pancreatic cancer, NSC-631570 alone and in combination with gemcitabine nearly doubled the median survival times in patients suffering from advanced pancreatic cancer. PMID- 11914933 TI - Efficacy of oral UFT as adjuvant chemotherapy to curative resection of colorectal cancer: multicenter prospective randomized trial. AB - BACKGROUND: The purpose of this study is to evaluate the efficacy of postoperative adjuvant chemotherapy using uracil and tegafur (UFT) for colorectal cancer. METHODS: In a multicenter trial among 43 institutions for patients who underwent curative resection of Dukes' B or C colorectal cancer, a surgery alone group (control group) and a treatment group (UFT group) to which UFT was administered at 400 mg/day for 2 years following surgery were compared. A total of 320 patients were registered between March 1991 and April 1994, and 289 of these patients were analyzed as a full-analysis set. RESULTS: The 5-year disease free survival rate was 75.7% in the UFT group and 60.1% in the control group, respectively, and the stratified log-rank test showed the statistical significance ( P=0.0081). This difference was marked in rectal cancer ( P=0.0016) and, in particular, the local recurrence was reduced. No significant difference was observed in the 5-year survival rate. The incidence of adverse reactions on administration of UFT was low, and there was no serious adverse reaction. CONCLUSION: It is suggested that the consecutive administration of UFT at 400 mg/day was an effective and highly safe therapeutic method as postoperative adjuvant chemotherapy for rectal cancer. PMID- 11914934 TI - A laparoscopic approach for the treatment of lower third esophageal diverticula. AB - Diverticula in the middle and lower thirds of the esophagus are a rare disorder, frequently associated with esophageal motility disturbances. For symptomatic patients transthoracic diverticulectomy with or without Heller's myotomy is recommended. Here we report two cases of symptomatic esophageal diverticula which were both treated successfully by a laparoscopic transhiatal approach. PMID- 11914935 TI - Catalytic activity of phospholipase A2 in blood plasma and pancreatic tissue perfusion in early experimental acute pancreatitis. AB - BACKGROUND: To investigate the sequence of changes in the catalytic activity of phospholipase A2 in plasma and pancreatic tissue perfusion and oxygenation in mild and severe acute pancreatitis in pigs. METHODS: Twenty-four pigs were randomized into the groups of severe acute pancreatitis, mild acute pancreatitis, and controls. The pancreatic duct of eight anesthetized and mechanically ventilated pigs was cannulated, and taurocholic acid was infused into the pancreatic duct to induce severe acute pancreatitis. Eight animals received intraductal saline and developed mild acute pancreatitis. Eight pigs were cannulated only and served as controls. RESULTS: Central hemodynamics, arterial blood gases, and acid-base balance were stable throughout the study period in all three groups. Pancreatic tissue oxygenation decreased in pigs with severe acute pancreatitis and increased in animals with mild acute pancreatitis. The catalytic activity of phospholipase A2 in plasma remained stable, and there was no difference between the groups. Similarly, C-reactive protein values remained within the normal range during the study period in all groups. CONCLUSION: Plasma phospholipase A2 levels do not react to the changes in pancreatic tissue perfusion in the early phase of mild and severe acute pancreatitis. PMID- 11914936 TI - Abdominal wall and foot reconstruction after extensive desmoid tumor resection with free tissue transfer. AB - BACKGROUND: Desmoid tumors are rare connective tissue tumors, also referred to as deep, aggressive fibromatosis. Although histologically benign, they show an invasive growth behavior and have a high local recurrence rate. METHODS: The treatment of choice is surgical resection with wide negative margins, while the use of radiotherapy remains controversial. Wide resection of greater desmoid tumors may result in considerable defects and functional impairment. Few papers discuss different options for defect coverage after desmoid tumor resection. Two cases of extensive desmoid tumors, one at the trunk, one at the foot, both with compromised wound margins due to multiple previous surgeries, are presented. To achieve a stable and functional soft tissue cover, the defects were treated with microvascular soft tissue transfer (one free latissimus dorsi, one free radial forearm flap). RESULTS: Both flaps healed uneventfully and patients regained full function of the abdominal wall and foot, respectively. CONCLUSIONS: The presented cases demonstrate the efficacy of free flap coverage as an ultimate therapeutic option in selected cases of critical defects after extra-abdominal desmoid tumor resection. Furthermore, free flaps provide a well vascularized ground for adjuvant radiotherapy. PMID- 11914937 TI - Embryonic development in the primitive bilaterian Neochildia fusca: normal morphogenesis and isolation of POU genes Brn-1 and Brn-3. AB - Neochildia fusca is a member of the taxon Acoela, a group of flatworms that, according to some recent molecular phylogenetic analyses, are distinct from other flatworms and constitute a basal branch with a sister taxon relationship to the rest of the Bilateria. In this paper, we analyze early neural development in this species and report the sequence and expression of two Pit-Oct-Unc (POU) genes, NeocBrn-1 and NeocBrn-3. Homologs of these highly conserved genes play a role in neural fate determination in vertebrates, Drosophila and Caenorhabditis elegans. Acoels, including Neochildia, have a unique invariant pattern of early cleavage called duet spiral cleavage. In subsequent cell divisions descendants of the first three micromere duets form an outer layer of epidermal and neural progenitors surrounding the meso/endoderm progenitors, which are themselves descended from the macromere duet 4A, B and the micromere duet 4a, b. Organ formation begins at mid-embryonic stages with the epidermal primordium adopting a ciliated epithelial shape. Sub-epidermally, a bilaterally symmetric brain primordium can be seen at the anterior pole. Laterally and posteriorly, myoblasts form a thin layer underneath the epidermis. In late embryos and juveniles of Neochildia, the brain is formed by a 3-4 cell-diameter-thick layer of neurons forming a cortex surrounding a neuropile that is relatively free of cell bodies. A highly regular "orthogonal" array of muscle fibers penetrates the brain. We have isolated and partially sequenced homologs of the vertebrate Brn-1 and Brn-3 genes, which we call NeocBrn-1 and NeocBrn-3, respectively. These sequences contain and span portions of the POU-specific domain and a homeodomain, and are sequence similar to their respective homologs in vertebrates and Drosophila. RT PCR reveals that NeocBrn-1 and NeocBrn-3 are expressed from mid-embryonic to adult stages. Whole-mount in situ hybridization shows expression of both genes in distinct subsets of nerve cells in juvenile and adult worms. NeocBrn-1 also appears in a subset of intra-epidermal gland cells. These observations are an initial step towards reconstructing the neural development of a key group of bilaterians, the Acoela. These flatworms, by virtue of their distinct morphology, development and phylogenetically basal placement, are likely to provide key insights into the interpretation of the evolution of metazoan neural architecture. PMID- 11914938 TI - HSP90 function is required for morphogenesis in ascidian and echinoid embryos. AB - Treatment of embryos of the ascidians Boltenia villosa and Cnemidocarpa finmarkiensis and the sea urchin Strongylocentrotus purpuratus with the anti HSP90 drugs geldanamycin and radicicol caused morphogenetic arrest. All embryonic stages during which obvious morphogenesis was observed were sensitive to treatment, including formation of the sea urchin blastular epithelium. Arrested embryos were viable for many hours to days post-treatment, indicating a low general toxicity of these drugs. Morphogenetic movements including gastrulation and migration (but not ingression) of sea urchin primary and secondary mesenchyme cells were arrested 8-10 h after treatment began. Cell division and developmentally regulated expression of some genes continued after morphogenesis was arrested. Anti-HSP90 drugs cause selective inactivation or degradation of proteins with which the protein chaperone HSP90 interacts. Therefore, morphogenetic arrest subsequent to the disruption of HSP90 function may result from the reduction in concentration, or activity, of client proteins required for morphogenetic movements of cells. The use of these drugs may provide a means to identify novel activities or proteins involved in morphogenesis. PMID- 11914939 TI - Expression cloning in ascidians: isolation of a novel member of the asctacin protease family. AB - The small genome size and gene number of ascidians makes them an ideal model system in which to screen for conserved genes that regulate the development of chordates. Expression cloning has proven to be an effective strategy for isolating genes that play a role in embryogenesis. We have taken advantage of the large size and ease of manipulation of Xenopus embryos for use as an assay system to screen for developmental regulatory genes from the ascidian Ciona intestinalis. Many invertebrate genes have been shown to function in vertebrates, providing us with precedent for our cross-species analysis. The first clone isolated from this screen is an astacin class metalloprotease. This ascidian astacin, named no va, causes a gastrulation defect in Xenopus. In C. intestinalis, no va is expressed both maternally and zygotically. The zygotic expression is seen in the mesenchyme of gastrula and neurula staged embryos. PMID- 11914940 TI - macho-1-Related genes in Ciona embryos. AB - In ascidians, maternal factor(s) localized in the myoplasm of the egg are essential for specification and subsequent differentiation of larval muscle cells. The macho-1 gene of Halocynthia roretzi encodes a zinc-finger protein: the gene is only expressed maternally, the resultant maternal mRNA is localized to the myoplasm, and the gene function is essential for muscle cell differentiation. Here we have characterized macho-1 homologues, Ci-macho1 of Ciona intestinalis and Cs-macho1 of Ciona savignyi. Interestingly, we found that the Ciona macho-1 genes are expressed both maternally and zygotically: their maternal transcript is localized to the myoplasm while their zygotic expression is seen after neurulation in cells of the central nervous system. Functional suppression of Cs macho1 with morpholino antisense oligonucleotide resulted in inhibition of the initiation of zygotic expression of a muscle-specific actin gene. We propose a possible evolutionary scenario in which an ancestral Zic-related gene gave rise to both the macho-1-like muscle determinant gene as well as neuronal Zic genes. PMID- 11914941 TI - Green fluorescent protein as a convenient and versatile marker for studies on functional genomics in Drosophila. AB - Gene function can be deduced from lack or gain of activity. For the manipulation of gene doses or activity in Drosophila, a set of P-based vectors was constructed containing green fluorescent protein as marker. pBLUEi, pGREENi and pYELLi were designed for large insert transformation. Mosaicism was generated in vivo with pFlipG which is also ideal for targeted gene disruption. Tissue-specific gene silencing in vivo was performed with the vector set pHIBS and pUdsGFP. pHIBS allows easy cloning and shuttling of double-headed constructs. With pUdsGFP, double stranded RNA can be produced in defined patterns and the area of interference simultaneously visualized by green fluorescence. We demonstrate nearly complete silencing of a ubiquitously expressed gene in a tissue-specific manner. PMID- 11914942 TI - Induction of gametogenesis in the basal cnidarian Nematostella vectensis(Anthozoa). AB - A protocol was established to reproducibly induce spawning in the basal cnidarian Nematostella vectensis (Anthozoa). We found that a combination of feeding regime, dark-light cycle and temperature shift synergistically induced gametogenesis in adult polyps. Females lay between 100-600 eggs. This procedure led reproducibly to the production of thousands of eggs over the course of more than 1 year in weekly cycles. Gametes are released in a time window of about 2 h resulting in predictable and fairly synchronized development. We also present a method for in vitro fertilization allowing manipulation of early embryos. These methods as well as the simple culture conditions could provide important prerequisites for the use of Nematostella as a model system for the development of a basal Metazoa. PMID- 11914943 TI - Mbx, a novel mouse homeobox gene. AB - The homeobox gene Mbxis thought to play important roles in the development of eyes and tectum in zebrafish. We isolated mouse Mbx cDNA and analyzed its expression pattern during early mouse embryogenesis. Expression is predominantly restricted to the midbrain region at E9.5. At subsequent stages of development, Mbx transcripts were also found in the forebrain in addition to midbrain. Thus, the Mbx gene provides a useful molecular marker for early mouse midbrain development and may play a critical role in brain development. PMID- 11914944 TI - [Diagnosis and therapy of stapes fractures and luxations]. AB - BACKGROUND: Injuries of the ossicular chain have various etiologies. Skull traumas from blows to the temporal, parietal or occipital region with or without fracture of the temporal bone are the main causes of the ossicular injury; other modes of injury are rare. It depends on the type of trauma which kind of injury occurs. Mainly subluxations, luxations and fractures are to be observed. The incus is the ossicle which is the most often injured. Mostly, only one ossicle is injured, rarely two or all of them. PATIENTS: From January 1997 to June 1999 we operated 19 patients with traumatic lesions of the membrane and the middle ear. In 4 cases the reason for the operation was an injury of the stapes. We observed 2 cases with broken stapes crura, 1 case with an impression of the stapes into the vestibulum with perilymphatic fistula and 1 case with subluxation of the stapes in the direction of the promontorium. By presenting 4 cases we want to point out the diagnostic possibilities and the operative management of such rare injuries. In all cases we applied the transmeatal approach for the operation of the middle ear. In 2 cases it was possible to place the stapes back in their anatomic position in order to re-establish the ossicular chain. In the other 2 cases we had to use middle ear prostheses which were interpositioned between the manubrium of the malleus and the footplate of the stapes. RESULTS: In all cases the immediate post-operative hearing results and the long-term hearing results after 10 to 28 months were satisfying. We did not notice any post-operative complications, especially no vestibular symptoms. One patient's tinnitus was the same as before the operation. CONCLUSIONS: The diagnosis of a stapes injury is despite the use of sophisticated technology, especially the CT-scan of the temporal bone, often very difficult. A sensitive anamnesis seems to be very important. It depends on the kind of injury which operative technique is to be applied in order to re-establish the ossicular chain. In our opinion, it is useful to take - if it is possible - the autolog stapes for the surgical re establishment of transmission. PMID- 11914945 TI - [Morphological changes of nasal and duodenal mucosa in patients with cystic fibrosis]. AB - BACKGROUND: Cystic fibrosis (CF) is an inherited multisystemic disorder that results in generalized dysfunction of exocrine glands. In patients with cystic fibrosis dyscrinia with affection of exocrine glands function is a main problem of the upper and lower respiratory tract. In addition to chronic obstructive pulmonary disease, chronic sinusitis, nasal polyposis and hypertrophy of inferior turbinates with nasal airway obstruction are typical signs. To understand pathophysiological mechanisms in CF and to correlate morphological findings with clinical symptoms, investigations of nasal mucosa are important. METHODS: Tissue samples of inferior turbinates were taken during nasal surgery from 7 children, ranging from 3 to 11 years of age between September 1998 and May 2000. Histological sections were cut followed by a light- and electron microscopical examination (EM 902 A Zeiss). Additionally, specimens of duodenal mucosa were investigated. RESULTS: In comparison with sections of normal nasal mucosa the lamina propria mucosae shows different morphological changes. Under a thick layer of respiratory epithelium with a high portion of goblet cells and particulary vacuoles there is an edematous subepithelial area. The capillary layer is reduced and the seromucous glands show an atypical morphological structure with widely mucous cells and cystic dilatation. On an ultrastructural level the glandular cells show atypical and inhomogeneous glandular droplets in the supranuclear cell portion. A viscous secretion was detectable at the glandular lumen. The nucleus contains dispersed chromatin as a sign of increased activity and the structures of Golgi apparatus were obviously detectable. CONCLUSIONS: In respective literature studies on the different morphological changes on light- and electron microscopical level in CF-associated rhinopathies are rare. This histological study demonstrated various morphological changes of nasal mucosa and shows a correlation between the glandular dysfunction and the typical symptoms in CF. Additionally a comparison with ultrastructural findings of CF-enteropathies is proposed. These findings could help to look at new aspects in the pathophysiology for patients with CF. PMID- 11914946 TI - [Long term results in adenoidcystic carcinoma]. AB - BACKGROUND: Due to the discreet initial symptoms and the locally aggressive infiltration with perineural spread the adenoidcystic carcinoma (ACC) presents a special diagnostical and therapeutical challenge. PATIENTS: : In a retrospective study the forms of 54 patients were analysed, whose average age was 55.5 (24 - 77) years. RESULTS: The sex ratio showed a slight female preponderance with 57 % to 43 %. The major salivary glands were affected in 26 cases. The exact distribution of the ACC was: parotid gland (n = 18), submandibular gland (n = 8), oral cavity (n = 10), paranasal sinuses (n = 11), nasopharynx (n = 4) and larynx (n = 3). The most common symptoms were a tumor-related swelling and pain which persisted for a duration of several months after final diagnosis could be established. Facial palsy was observed in 4 patients. Histological examination revealed a tubular subtype in 4 cases, in 28 cases a cribriforme and in 12 cases a solid subtype. No definitive differentiation was possible in 10 specimens. The skull base was infiltrated in 16 patients. Except one patient all 54 underwent surgical therapy. Postoperative radiotherapy was additionally given in 25 cases which was combined with a chemotherapy in 6 patients. Nevertheless ACC recurred in 60 % of our patients. Lymphnode metastases were observed in 13 patients after a latency of 3.3 years in average, but predominantly pulmonary metastases as distant spread developed in 18 patients after 5.8 years significantly later. The overall survival rate was 84.38 % after 2 years, 75.90 % after 5 years, 50.49 % after 10 years and 20.11 % after 20 years. Male sex, infiltration of the skull base and histological evidence of perineural and perivascular spread proved to be statistically significant factors for an unfavourable prognosis. CONCLUSIONS: Due to the uncommon biological behaviour with a slow growth on the one hand side and an aggressive local invasion on the other hand side the ACC can be regarded as a challenging malignant disease for the clinician whose adequate therapy does not allow any standardized regime. The tendency for recurrence even after a period clinically free of symptoms makes a life long follow-up mandatory. PMID- 11914947 TI - [Brachytherapy with (192)Iridium in the treatment of recurrent nasopharyngeal carcinoma]. AB - BACKGROUND: Local recurrence of nasopharyngeal carcinomas can be treated in different ways. One option is re-irradiation e. g. as stereotactically guided convergent beam. An operative approach is possible in small recurrent tumours. Brachytherapy is a good alternative because of the steep dose gradient in the pre irradiated area. METHODS: First step is the creation of a wide approach to the nasopharynx. This leads to a tumour mass reduction and gives space for the applicator. The radioactive substance will be brought into contact with the tumour by an afterloading procedure. The advantage of this therapy is the possibility to protect the surrounding tissue whereas the tumour receives a relatively high dosage. Between 2/90 and 12/96 10 men and 3 women were treated according to this protocol. RESULTS: The median age was 56 years (37- 66 years), the average follow-up period was 49,7 months (8 -123 months). 2 non keratinising and 11 undifferentiated carcinomas were treated. 5 of 13 patients were still alive at the end of the follow-up period. The 5 year over all survival rate was 46 %. 3 patients are free of disease for 5 years or longer. The patients were treated 2 to 6 times with 7 Gy in 5 mm depth. CONCLUSIONS: The results show that good palliation can be achieved by the applied method but larger studies are required to give a definite statement. PMID- 11914949 TI - [The tracheostomy-sphincter procedure - case report of a modified tracheostomy]. AB - BACKGROUND: For an adequate speech production after tracheostomy patients are dependant on a temporary closure of the stoma. This closure can either be done manually or by the use of various tracheostoma valves. The tracheostomy-sphincter procedure, which is introduced in this case, allows the voluntary constriction of the stoma. METHODS: This case describes the treatment of a paediatric patient with laryngeal stenosis. During tracheostomy formation, an artificial muscle sphincter was created, using the sternohyoid muscle (strap muscle). RESULTS: Postoperative contractile movements were possible, which were improved by special exercises. After 3 months the complete closure of the tracheostoma was achieved. CONCLUSIONS: The establishment of a tracheostomy-sphincter can be carried out during tracheostomy and allows the patient to speak without the use of a device. Tracheobronchial secretions can be cleared via the pharynx by coughing. PMID- 11914948 TI - [Chronic myoepithelial sialadenitis - symptomatology, clinical signs, differential diagnostics]. AB - In the differential diagnosis of mass lesions of the salivary glands, myoepithelial sialadenitis (MESA), i. e. benign lymphoepithelial lesion, carries particular importance because of its association with Sjoegren's syndrome and development of malignant lymphoma. In the present study, epidemiology and clinical findings were analysed in relation to presence of MESA, Sjoegren's syndrome and lymphoma development. MATERIAL AND METHOD: 67 patients, histopathologically classified by the salivary gland registry, were analysed retrospectively in regard to their clinical presentation, especially in regard to the chronical process of inflammation as present in MESA. RESULTS: MESA primarily affects women in the 5th and 6th decade and regularly the parotid gland; in 44.8 % of the cases, there is multiple organ presentation. Xerostomy (38.5 %) is usually present (in 88,9 % of all cases) before or at clinical onset of gland inflammation, whereas xerophthalmy (28.4 %) did not show such a correlation. In general, rheumatic diseases (23.9 %) precede the gland-symptoms in 77.8 % of the patients. In 31.3 % of the cases a Sjoegren's syndrome was present. 26.9 % of the patients developed a malignant Non Hodgkin's Lymphoma (88.9 % of the MALT-type). CONCLUSION: The most important clinical relevance of MESA lies in the higher probability to develop malignant lymphoma; this requires adequate staging procedures and proper histopathological examination of sialogenic and nodal masses, especially over the course of the disease. PMID- 11914950 TI - [The monthly interesting case No. 52. Plexiform ameloblastoma of the maxillary sinus]. PMID- 11914951 TI - [State of the Art. Diagnostic imaging of paranasal sinus diseases]. AB - The following article describes a diagnostic concept of paranasal sinus imaging, based on more than 10 years close collaboration of the authors. CT and MR are the primary imaging modalities for investigating paranasal sinus lesions. Conventional X-ray images add little information due to insufficient sensitivity. Angiography is restricted to those patients who require preoperative tumor embolization or emergency treatment of intractable epistaxis. After thorough medical treatment of inflammatory paranasal sinus disease, CT becomes the method of choice to clearly depict preoperative osseous anatomy and to distinguish between benign and malignant intraosseous lesions. In cases where trauma is involved, CT easily identifies anterior skull base fractures. Moreover, CT will be the first step to localize early or late CSF rhinorrhea. MR, with its ability to distinguish between different soft tissues, compliments CT. This holds particularly true for inflammatory or neoplastic disease close to the skull base, orbital apex and cavernous sinus. The signal intensity in T(1) and T(2) weighted images provides additional information with respect to cellularity and vascularization of lesions, which has important diagnostic and therapeutic implications. PMID- 11914952 TI - Antioxidant and photoprotective activity of a lipophilic extract containing neolignans from Krameria triandra roots. AB - The antioxidant/photoprotective potential of a standardized Krameria triandra (KT) root extract (15% neolignans) has been evaluated in different cell models, rat erythrocytes and human keratinocytes cell lines, exposed to chemical (cumene hydroperoxide, CuOOH) and physical (UVB radiation) free radical inducers. The extract was significantly more active (IC50 0.28 +/- 0.04 microg/ml) than the typical chain-breaking antioxidant alpha-tocopherol (IC50 = 6.37 +/- 0.41 microg/ml) in inhibiting the CuOOH-induced hemolysis in rat blood cells. The KT constituent 2-(2,4-dihydroxyphenyl)-5-(E)-propenylbenzofuran, was the most active (IC50 = 0.03 +/- 0.005 microg/ml), followed by eupomatenoid 6 (IC50 = 0.29 +/- 0.06 microg/ml) and conocarpan (IC50 = 0.77 +/- 0.08 microg/ml). The same order of potency was observed in red blood cells exposed to UVB irradiation in continuo, with IC50 values 0.78 +/- 0.08 microg/ml for KT extract, 0.18 +/- 0.02 microg/ml for 2-(2,4-dihydroxyphenyl)-5-(E)-propenylbenzofuran, 0.95 +/- 0.11 microg/ml for eupomatenoid 6, and 3.8 +/- 0.39 microg/ml for conocarpan. In cultured human keratinocytes exposed to UVB radiation (50 mJ/cm2), KT extract (2.5-20 microg/ml) significantly and dose-dependently restrained the loss in cell viability and the intracellular oxidative damage: glutathione (GSH) depletion and the rise in dichlorofluorescein (DCF), marker of peroxide accumulation, were suppressed by 20 microg/ml KT and in parallel cell morphology maintained. The cytoprotective effect of the extract was confirmed in a more severe model of cell damage: exposure of keratinocytes to higher UVB doses (300 mJ/cm2), which induce a 50% cell death. In keratinocyte cultures supplemented with 10 microg/ml, cell viability was almost completely preserved and more efficiently than with (-) epigallocatechin 3-gallate and green tea. The results of this study indicate the potential use of Rhatany extracts, standardized in neolignans, as topical antioxidants/radical scavengers against skin photodamage. PMID- 11914953 TI - Mitochondrial destabilisation and caspase-3 activation are involved in the apoptosis of Jurkat cells induced by gaudichaudione A, a cytotoxic xanthone. AB - Gaudichaudione A, a cytotoxic xanthone obtained from Garcinia gaudichaudii, displayed strong growth inhibitory effects against Jurkat human leukemic cells, parental murine leukemic P388 and P388/DOX-resistant cell lines, and exhibited similar cytotoxicity against P388/DOX as well as P388, but was less toxic toward normal human Chang liver cells. A variety of apoptotic events induced by gaudichaudione A at the mitochondrial and nuclear levels has been characterized. It appears that gaudichaudione A induced a collapse of the mitochondrial transmembrane potential. Caspase-3 was activated and induced internucleosomal DNA fragmentation and externalization of PS residues. The apoptotic features were also observed in ultrastructural studies. PMID- 11914954 TI - Cytotoxic activity of sesquiterpenoids from Atractylodes ovata on leukemia cell lines. AB - The rhizome of Atractylodes ovata (Bai Zhu in Chinese) is a widely used traditional Chinese herb in Taiwan as a tonic agent. In this paper, four sesquiterpenoids, namely atractylon, and atractylenolides I, II, and III, were isolated from the n-hexane extract of A. ovata and were evaluated for cytotoxic effects in vitro. Atractylon significantly inhibited the growth of human leukemia cell line HL-60 and mouse leukemia cell line P-388, and showed low cytotoxicity against primary cultures of normal human peripheral blood mononuclear cells at 15 microg/ml for 12 h. Atractylon had a dose-dependent antiproliferative effect on the two tumor cell lines. In accordance with DNA fragment increases and PARP protein decreases, atractylon at 15 microg/ml for 6 h induced apoptosis in HL-60 cells. Moreover, atractylon inhibited the viability of P-388 cells and induced apoptosis after 15 microg/ml treatment for 12 h in an in vitro assay. However, atractylenolide I at 30 microg/ml for 12 h also induced apoptosis in HL-60 and P 388 cells, but atractylenolides II and III showed no significant inhibition effects on tumor cell growth. As the above results suggested, atractylon and atractylenolide I were the major cytotoxic principle constituents of A. ovata on leukemia cell lines. PMID- 11914955 TI - Evaluation of the antiviral activity against Junin virus of macrocyclic trichothecenes produced by the hypocrealean epibiont of Baccharis coridifolia. AB - Four macrocyclic trichothecenes, roridin A, roridin E, verrucarin A and verrucarin J, produced by the hypocrealean epibiont of Baccharis coridifolia, were evaluated for their inhibitory activity against the arenavirus Junin (JUNV), the etiological agent of Argentine hemorrhagic fever. The trichothecenes achieved a dose-dependent inhibition of JUNV multiplication at concentrations not affecting cell viability. The 50 % inhibitory concentration (IC50) values determined by a virus yield inhibition assay were in the range 1.2 - 4.9 ng/ml. The most active compound was verrucarin J which reduced JUNV yield more than 2 log units and had a similar effect against the arenavirus Tacaribe. The trichothecenes lacked virucidal effects on JUNV virions. From time of addition and removal experiments, it can be concluded that verrucarin J inhibited a late stage in the replicative cycle of JUNV, after 5 h of adsorption. PMID- 11914956 TI - Muscle relaxing activity of Hyssopus officinalis essential oil on isolated intestinal preparations. AB - The muscle relaxing activity of the essential oil of Hyssopus officinalis L. (Lamiaceae) and some of its main components (isopinocamphone, limonene and beta pinene) was studied on isolated preparations of guinea-pig and rabbit intestine. The essential oil and isopinocamphone inhibited the acetylcholine- and BaCl2 induced contractions in guinea-pig ileum in a concentration-dependent manner (IC50 42.4 microg/ml and 61.9 microg/ml to acetylcholine; 48.3 microg/ml and 70.4 microg/ml to BaCl2) whereas limonene or beta-pinene left tissue contraction unchanged. In guinea-pig ileum H. officinalis essential oil also blocked the contractions induced by CaCl2. In isolated rabbit jejunum the essential oil reduced the amplitude of spontaneous movements and decreased the basal tone; neither haemoglobin, methylene blue, N(omega)-nitro-L-arginine methyl ester (L NAME) or propranolol blocked the myorelaxant effect. PMID- 11914957 TI - Schisandrin B protects against tacrine- and bis(7)-tacrine-induced hepatotoxicity and enhances cognitive function in mice. AB - Intragastric administration (100-200 micromol/kg) of tacrine (THA) or bis(7)-THA could cause an acute and dose-dependent increase in plasma alanine aminotransferases activity in mice at 6 h after the drug administration. The increase in plasma enzyme activity was associated with an increase in hepatic malondialdehyde level, an indirect index of oxidative tissue damage. Pretreating mice with schisandrin B (Sch B), an active dibenzocyclooctadiene derivative isolated from the fruit of Schisandra chinensis, at a daily dose of 0.125-0.5 mmol/kg for 3 days protected against the THA/bis(7)-THA induced hepatic oxidative damage in a dose-dependent manner. Sch B treatment (0.025-0.5 mmol/kg/day x 5) also enhanced the passive avoidance-response in mice as assessed by the step through task experiment. The ensemble of results suggests that Sch B may be useful for reducing the potential hepatotoxicity of THA/bis(7)-THA in anti Alzheimer's therapy. PMID- 11914958 TI - Platycodin D and D3 increase airway mucin release in vivo and in vitro in rats and hamsters. AB - The root of Platycodon grandiflorum has been widely used for the treatment of various chronic inflammatory diseases including airway disease in oriental medicine. The root extract of the plant has been known to be effective in the expectoration of sputum or mucus, thereby improving airway respiratory function and preventing secondary airway inflammation. In this study, we investigated the effect of platycodin D and D3, the saponin components that are anti-inflammatory components in Platycodon grandiflorum. Platycodin D and D3 increased mucin release from rat and hamster tracheal surface epithelial cell culture and also from intact rat trachea upon nebulization. The effect of platycodin D3 was stronger than that of ATP, a potent mucin secretagogue and also of ambroxole, a mucolytic drug. The results from the present study suggest that platycodin D and D3 are useful as expectorant agents in the treatment of various airway diseases. PMID- 11914959 TI - Local variation in the content of angiotensin II and arginine vasopressin receptor antagonistic terpenoids in the rhizomes of Alisma orientale. AB - The aqueous extract of the Chinese crude drug Alismatis rhizoma, a dried rhizome of Alisma orientale Juzepczuk, exhibited in vitro inhibitory activities on angiotensin II and arginine vasopressin binding to their receptors. A fractionation study on the extract clarified that the known terpenoidal constituents; i.e., alisols A and B, 23-O-acetylalisol B, and alismol, were responsible for the activities. Furthermore, investigation of commercial samples of the crude drug demonstrated striking differences in their activities dependent on their locality of origin due to a difference in the amounts of these active principles. Therefore, the content of these principles could be utilized as a criteria of quality of the crude drug Alismatis rhizoma. PMID- 11914960 TI - An arabinogalactan-protein from cell culture of Malva sylvestris. AB - An arabinogalactan-protein (AGP) from suspension culture medium of Malva sylvestris was isolated by precipitation with beta-glucosyl Yariv reagent, followed by gel-permeation chromatography. It revealed characteristic features of AGPs: a high amount of polysaccharide with a ratio of galactose to arabinose of 1.9 : 1, some uronic acids, and a small protein moiety with the main amino acids serine, alanine and hydroxyproline. The molecular weight was estimated to be 1.3 x 10(6) Da. Linkage analyses showed that the AGP is composed of a highly branched core polysaccharide of 3-, 6-, and 3,6-linked Galp residues with terminal Araf, GlcAp and Galp. Partial acid hydrolysis resulted in loss of Araf residues at the periphery of the molecule and heavily reduced its reactivity with beta-glucosyl Yariv antigen. PMID- 11914961 TI - Structural features of an immunostimulating and antioxidant acidic polysaccharide from the seeds of Cuscuta chinensis. AB - Three crude polysaccharide fractions, named CS-A, CS-B and CS-C, were prepared from the seeds of Cuscuta chinensis by hot water extraction and diluted alkali extraction subsequently, then EtOH precipitation, and tested for lymphocyte proliferation activity. CS-A showed a stimulating effect on concanavalin A or lipopolysaccharide induced mitogenic activity of lymphocytes. An acidic polysaccharide (CS-A-3beta) was purified from CS-A by anion exchange and gel filtration chromatography. The polysaccharide showed potent stimulating effects on lymphocyte proliferation and antibody production, but did not significantly influence the serum IgG level. Its structural features were elucidated by methylation analysis, partial acid hydrolysis, 1D and 2D NMR and ESI-mass spectroscopy. The data obtained indicated that CS-A-3beta had a backbone consisting of alpha-D-1,4-linked GalpA residues and beta-L-1,2-linked Rhap residues with branches at C-4 of Rhap residues and C-3 of GalpA residues, composed of arabinogalactan and glucobiose. This structure is typical for a pectic polysaccharide of the rhamnogalacturonan type. In addition, the effect of CS-A, CS-B, CS-C and CS-A-3beta on hydrogen peroxide induced cell lesion in rat pheochromocytoma line PC 12 was investigated. The results indicated that, besides its immunostimulating activity, CS-A-3beta had a protective effect against free radical-induced cell toxicity. PMID- 11914962 TI - New macrocyclic diterpenoids from Euphorbia esula. AB - The structures of two new macrocyclic jatrophane diterpenoid esters from the whole herb of Euphorbia esula, were established as 11,14-epoxy 3beta,5alpha,7beta,8alpha,9alpha,15beta-hexaacetoxy-12-oxo-13alphaH-jatropha 6(17)-ene (1) and 1alpha,3beta-diacetoxy-5alpha,7beta-dibenzoyloxy-9,14-dioxo 11beta,12alpha-epoxy-2alpha,8alpha,15beta-trihydroxy-13betaH-jatropha-6(17)-ene (2) by a combination of 1D- and 2D-NMR techniques as well as UV, IR and mass spectral data. Bioassay evaluation of all isolates against the human tumor cell lines (B16, KB, SMMC and BGC) indicated that ester 2 was cytotoxic to B16 with the IC50 value being 1.81 microg/ml. In addition, the irritant activity assay indicated that both diterpenoids were inactive (ID(24)50 > 100 microg/ear). PMID- 11914963 TI - Cytotoxic diterpenoids from Euphorbia pekinensis. AB - A new diterpenoid, named euphpekinensin, along with three known diterpenoids, was isolated from the roots of Euphorbia pekinensis for the first time and the structures were elucidated by spectral analysis. The 2D-NMR techniques such as 1H 1H COSY, HMQC, HMBC and NOESY spectra were mainly applied to determine the structure of the new diterpenoid. The four diterpenoids showed cytotoxic activity against human KB cells in vitro. PMID- 11914964 TI - Taxuyunnanines S-V, new taxoids from Taxus yunnanensis. AB - Four new 11(15-->1)-abeotaxoids with an opened oxetane ring system, taxuyunnanines S, T, U and V (1 - 4) were isolated from the bark of Taxus yunnanensis. Taxuyunnanine S (1), is the most hydroxylated abeotaxoid discovered to date from Taxus species. The structures were determined by spectroscopic means including 1D and 2D NMR experiments. PMID- 11914965 TI - Bisxanthones from Hypericum japonicum: inhibitors of PAF-induced hypotension. AB - The CHCl3-soluble part (HJ) of the MeOH extract from Hypericum japonicum afforded new bisxanthones, jacarelhyperols A (1) and B (2). The structures were elucidated by spectral methods. These compounds and HJ showed significant inhibitory effects against PAF-induced hypotension by an in vivo evaluation method, which can reveal the pharmacological action of the test material. PMID- 11914966 TI - Benzophenanthridine alkaloids from Corydalis flabellata. AB - Four new alkaloids, characterized as 6-(2-hydroxyethyl)-5,6-dihydrosanguinarine (1), 6-acetonyl-5,6-dihydrosanguinarine (2), N-methyl-2,3,7,8-tetramethoxy-5,6 dihydrobenzophenanthridine-6-ethanoic acid (3), N-methyl-2,3,7,8-tetramethoxy-6 oxo-5,6-dihydrobenzophenanthridine (4), together with oxosanguinarine (5), spallidamine (6), 6-acetonyl-5,6-dihydrochelerythrine (7), 6-oxochelerythrine (8) and sanguidimerine (9) were isolated from the roots of Corydalis flabellata. PMID- 11914968 TI - Evaluation of the anti-inflammatory effect of baicalein on dextran sulfate sodium induced colitis in mice. AB - The anti-inflammatory effect of three flavonoids from the root of Scutellaria baicalensis (baicalein, baicalin and wogonin) was evaluated in a murine model of acute experimental colitis induced by dextran sulfate sodium (DSS). Baicalein, but not baicalin or wogonin, given orally at 20 mg/kg for ten days, ameliorates all the considered inflammatory symptoms of the induced colitis, such as body weight loss, blood haemoglobin content, rectal bleeding and other histological and biochemical parameters. The effect of baicalein was similar to that of sulfasalazine, the reference drug given at 50 mg/kg. PMID- 11914967 TI - Effect of Mahonia aquifolium active compounds on interleukin-8 production in the human monocytic cell line THP-1. AB - The effect of the crude extract and of two alkaloid fractions prepared from Mahonia aquifolium on interleukin-8 (IL-8) production in the lipopolysaccharide (LPS)-stimulated human monocytic cell line THP-1 was studied. The production of IL-8 by cells stimulated with 20 ng/ml LPS after 48 h treatment with 20 microg/ml crude extract was inhibited by about 30 %. LPS-stimulated cells treated with 0.1 microg/ml bisbenzylisoquinoline alkaloid fraction exhibited a 40% inhibition of IL-8 production in comparison with control cells. The fraction of protoberberine alkaloids had no significant inhibitory activity. Weak or no inhibition of IL-8 production was found after treatment with 0.5 microg/ml of protoberberine alkaloids berberine and jatrorrhizine and with berbamine from the group of BBI alkaloids. In contrast, the production was inhibited after treatment with BBI alkaloids baluchistine (about 20%) and aromoline (up to 30%). PMID- 11914969 TI - Deoxypodophyllotoxin; the cytotoxic and antiangiogenic component from Pulsatilla koreana. AB - The petroleum ether fraction of Pulsatilla koreana (Ranunculaceae) was found to have an inhibitory effect on the tube-like formation of human umbilical venous endothelial (HUVE) cells and strong cytotoxic activity against five tumor cell lines. The active component isolated from the fraction was deoxypodophyllotoxin (DPT). The cytotoxic activity against the tumor cells comprising the A549, SK-OV 3, SK-MEL-2, HCT15, and B16F10 cell lines, expressed as ED50, ranged from 6 to 18 ng/ml. 3 ng/ml of DPT, a concentration considerably below the cytotoxic concentration, completely inhibited the tube-like formation of HUVE cells. Furthermore, DPT exhibited an inhibition ratio of 60 % on BDF1 mice bearing Lewis lung carcinoma cells. The inhibitory effect on the tube-like formation was suggested to play an important role in antitumor activity. PMID- 11914970 TI - Croblongifolin, a new anticancer clerodane from Croton oblongifolius. AB - A new furoclerodane, croblongifolin, together with one known clerodane, crovatin and one known labdane, nidorellol, were isolated from the stem bark of Croton oblongifolius. Structures were established based on spectroscopic and X-ray analysis. Croblongifolin showed a significant cytotoxicity against various human tumor cell lines including HEP-G2, SW620, CHAGO, KATO3 and BT474. PMID- 11914971 TI - Antimicrobial activity of 9-O-acyl- and 9-O-alkylberberrubine derivatives. AB - For the structure-activity relationship study on berberrubine derivatives, a series of compounds bearing 9-O-acyl- and 9-O-alkyl-substituents were synthesized and tested for antimicrobial activity against Gram-positive, Gram-negative bacteria and fungi. Octanoyl, decanoyl, lauroyl derivatives among the acyl analogs and hexyl, heptyl, octyl, nonyl, decyl, undecyl derivatives among the alkyl analogs showed strong antimicrobial activity against Gram-positive bacteria and fungi. As a whole, alkyl analogs were more active than acyl analogs for antimicrobial activity. Synthesized derivatives had no activity on Gram-negative bacteria. Too short or too long substituents decreased activity. These results suggest that the presence of lipophilic substituents with moderate sizes might be crucial for the optimal antimicrobial activity. PMID- 11914972 TI - Antispasmodic and antimicrobial diterpenic acids from Viguiera hypargyrea roots. AB - Two spasmolytic diterpene acids, ent-beyer-15-en-19-oic acid and ent-kaur-16-en 19-oic acid, have been isolated from the roots of Viguiera hypargyrea by bioassay guided fractionation. Beyerenoic acid was also active against Staphylococcus aureus, Enterococcus feacalis and Candida albicans. PMID- 11914973 TI - Antinociceptive constituents of Auricularia polytricha. PMID- 11914974 TI - Four highly oxygenated isopimarane-type diterpenes of Orthosiphon stamineus. AB - Four highly oxygenated isopimarane-type diterpenes, named orthosiphols O, P and Q and nororthosiphonolide A, have been isolated from the aerial parts of Orthosiphon stamineus from Myanmar, together with three known diterpenes, orthosiphols D and E and orthosiphonone A. Their structures were determined on the basis of extensive spectral analysis. All the isolated compounds displayed mild antiproliferative activities against highly liver metastatic colon 26-L5 carcinoma and human HT-1080 fibrosarcoma cell lines. PMID- 11914975 TI - Opioid receptor blockade eliminates mood effects of aerobic gymnastics. AB - The contribution of opioid receptors to the mood effects of aerobic gymnastics was tested by oral administration of naltrexone (25 or 50 mg) in 12 healthy women (aged 22 - 30 years). The opioid receptor blockade eliminated decreased anxiety, negative affect level and depressiveness as well as increased positive affect level, as found in a placebo trial after a 50 min session of aerobic gymnastics. 50 mg of naltrexone was more effective than 25 mg. Administration of 50 mg of naltrexone without exercise did not cause any significant changes in anxiety, positive and negative affect levels or depressiveness. PMID- 11914976 TI - Short-term effects of prolonged strenuous endurance exercise on the level of haematocrit in amateur cyclists. AB - Knowledge is sparse about the extent of potential dehydration due to prolonged strenuous cycling and its haematological acute effects on the haematocrit (Hct) in study populations credibly not taking any kind of doping. With increasing training load levels of Hct and haemoglobin (Hb) decrease in both amateurs and professionals as a long-term consequence due to expanded plasma volume (PV). On a short-term basis, however, counteracting dehydration potentially brought about by endurance exercise may cause a rise in Hct bringing competitive cyclists into conflict with the current condition regulations and Hct cut-off of 50 % set by the International Cycling Union (UCI) in its fight against erythropoietin (rhEPO) doping. On the other hand adequate and sufficient fluid substitution being substantial for a successful endurance performance should prevent any pronounced Hct rises. To study the haematological acute effects of prolonged strenuous cycling we measured Hct, Hb, red blood cell (RBC) count and plasma protein in a reliably 'clean' population of 38 well-trained male amateur cyclists before, immediately after and one day after an extraordinary ultramarathon. The pre-race levels of Hct, Hb and RBC count were placed in the lower range of normal distribution and well below the Hct cut-off limit of the UCI. Immediately post exercise the mean levels of Hct, Hb, RBC count and protein remained unchanged. One day after race, however, all four parameters significantly dropped by 3 %, 6.7 %, 6.5 %, 9.9 % respectively (p < 0.001), indicating marked post-exercise PV expansion. The calculated percentage increase in PV was 11.9 %. No evidence for coexisting exercise-induced haemolysis was found. Our study shows that in "clean, rhEPO-free" amateur cyclists who involve in strenuous marathon cycling the haematological short-term effects of extraordinary marathon cycling consist in considerable PV expansion making Hct values fall on the following day. The findings - gained from amateurs though - suggest that despite all its disadvantages the UCI Hct cut-off represents an appropriate means to discourage from excessive rhEPO doping at least as long as the available direct methods for detecting this kind of misuse are not yet applied by the international sports federations. PMID- 11914977 TI - Cardiorespiratory responses and blood lactate during an experimental run-cycle transition in duathletes. AB - The aim of this study was to determine the effects of a prior run on the cardiorespiratory responses measured during a subsequent cycle segment. Twelve duathletes underwent three successive laboratory trials at an interval of one week: 1) an incremental cycle test, 2) 20 min of running followed by 20 min of cycling (RC), and 3) 20 min of control cycling (C) at the same intensity as the cycling segment of RC. Ventilatory data were collected every minute using a breath-by-breath automated system. Blood samples were collected to measure venous blood lactate concentration, [La], at rest, after the running and cycling segments of RC and after C. The results showed that the C segment of RC had significantly higher VE, VE/VO2, f and HR than C alone and significantly lower VT (p < 0.05) than C alone. Moreover, steady state during C of RC was reached at the 2nd min for VO2, VE, VCO2, VE/VO2, VE/VCO2, and VdT; at the 4th min for R and HR, and at the 5th min for f. The C of RC induced a significant increase in [La] in comparison with C alone. We concluded that the first minute of cycling after running during an RC trial induced specific metabolic and cardiorespiratory responses. PMID- 11914978 TI - Growth status and estimated growth rate of young distance runners. AB - A mixed-longitudinal design was used to evaluate the growth status (size attained) and estimated growth rate of competitive young distance runners. Twenty seven male and 27 female distance runners were enrolled in the study based on race performance times. Ages ranged from 8.0 to 15.1 years at initial visit. Twenty males and 16 females were followed at approximate annual intervals for 4 or 5 years. Stature (cm) and body mass (kg) were measured on each occasion and annual growth velocities were estimated. Stature and body mass were plotted relative to age- and sex-specific U.S. reference data, while estimated velocities of stature (cm/yr) and body mass (kg/yr) were plotted relative to age- and sex specific percentiles from the Zurich Longitudinal Study. Differences in growth pattern between the runners and the reference values were compared statistically with a growth curve estimation regression model. Mean statures of male and female distance runners approximate the reference medians, while mean body masses are slightly below reference medians. Mean stature and body mass also maintain a constant position relative to the reference values in both sexes over time. Estimated velocities for stature and body mass are, on average, similar to age- and sex-specific medians for Swiss youth. Slope values indicate no significant difference in growth patterns between young distance runners and reference values. The findings are consistent with other data indicating that intensive endurance training during childhood and adolescence does not influence size attained and rate of growth in stature and body mass. PMID- 11914980 TI - Long-term vigorous training in young adulthood and later physical activity as predictors of hypertension in middle-aged and older men. AB - 500 and 69 male former elite athletes and 319 male controls completed a health questionnaire in 1985 and in 1995. Register data on the subjects were also collected. Subjects were aged 65 years or less and had no history of hypertension in 1985, and they had been healthy at the age of 20 years. The athletes were grouped into endurance and mixed sports (n = 386), and power sports (n = 183). The cumulative 10-year incidence of hypertension up to 1995 was significantly lower in the endurance and mixed sports group (23.6 %) compared to the power sports group (33.3 %) or the control group (32.0 %). The difference between the endurance and mixed sports group and the two other groups was still significant after adjustment for age, but not after further adjustment for body mass index, alcohol consumption, and later physical activity. However, the trend of reduced risk remained. In conclusion, a history of being an elite athlete in endurance or mixed sports predicts a lower risk of hypertension in working age men, while a history of being an elite athlete in power sports appears to confer no benefit. Later physical activity was also associated with lower risk. PMID- 11914979 TI - Alactic anaerobic performance in subjects with sickle cell trait and hemoglobin AA. AB - The aim of the present study was to assess the performance of subjects with sickle cell trait (SCT) during brief and explosive exercise involving mainly anaerobic metabolism. One hundred and ninety-six black subjects underwent SCT screening, which revealed the presence of 16 subjects with SCT and 180 subjects with normal hemoglobin (HbAA). All subjects performed four tests: 1) a 100-m sprint, 2) a long-jump, 3) a Leger-Boucher shuttle test and 4) a jump-and-reach test. A control group (n = 18) selected from the 180 subjects with HbAA was matched according to the sex, age, weight and height of the SCT subjects (SCTs). The performances of the SCTs (n = 16) were compared with those of the control group. The performances were similar between the SCTs and control group for the sprint test, long-jump and the Leger-Boucher shuttle test. There was, however, a significant difference for the jump-and-reach test between the two groups: the SCTs (i. e., males plus females, and males and females considered separately) reached a significantly greater height (p < 0.05) than the matched subjects of the control group (63.7 +/- 3.6 vs. 58.6 +/- 3.1 cm, 72.3 +/- 3.9 vs. 67.1 +/- 2.4 cm and 52.7 +/- 3.2 vs. 45.3 +/- 2.0 cm for SCTs versus non-SCTs, for the group, the males and the females, respectively). The results of the present study suggest that the performance of brief and explosive exercise may be enhanced by HbS. PMID- 11914981 TI - Quantitative effects of respiration on venous return during single knee extension flexion. AB - Respiration and the muscle pump play major roles in increasing venous return. However, the relative contribution of each of these factors remains unclear. The present study investigates the quantitative effects of interaction between respiration and the muscle pump on femoral venous blood flow (FVBF) during a single voluntary knee extension-flexion (KEF) using duplex-Doppler ultrasound. During various respiration modes, which consisted of arrested respiration, normal respiration and deep respiration (inspiration or expiration), eight subjects performed a supine one-legged voluntary KEF. FVBF was measured during respiration only (Protocol A) and during KEF synchronized with respiration (Protocol B). The difference between FVBF values obtained in Protocol B and Protocol A was defined as DeltaFVBF. When KEF was synchronized with normal or deep respiration, FVBF with inspiration was significantly lower than that with expiration. However, DeltaFVBF was significantly higher with inspiration than with expiration during deep respiration but was not significant during normal respiration. Furthermore, DeltaFVBF was significantly higher at both normal and deep respiration than at arrested respiration. The effects upon the venous return during KEF differed between inspiration and expiration. The present findings indicate that during a single supine KEF, respiration might promote venous return to a range of 1.5- to 2.3-fold DeltaFVBF during arrested respiration. PMID- 11914982 TI - Relationships between aerobic energy cost, performance and kinematic parameters in roller ski skating. AB - The aims of this study were to test the hypotheses that 1) the aerobic energy cost of roller ski skating (CS) is significantly related to level of performance and 2) a significant part of inter-individual differences of CS can be explained by kinematic parameters of skating locomotion. Oxygen uptake, kinematics of the knee and ankle joint, EMG of the vastus lateralis (VL) and gastrocnemius lateralis (GL) muscles, and roller ski velocity were recorded in 13 skiers who roller skied at 19.0 +/- 0.1 km x h(-1). CS was found to be 2.51 +/- 0.35 J x kg( 1) x m(-1) and significantly correlated with the skiers' level of performance (r = 0.61; p < 0.05). Significant relationships were found between CS and 1) knee angular amplitude (r = 0.75; p < 0.01) during the concentric phase, 2) VL average EMG during the concentric phase (r = 0.72; p < 0.01) and 3) VL and GL average EMG during the eccentric phase (p < 0.05). The results of this study showed that a significant part of performance could be explained by the aerobic energy cost of locomotion in skating. It can also be suggested that differences in upper/lower body utilization and/or in mechanical efficiency may explain the differences in CS. PMID- 11914983 TI - Simple measurements in assessing muscle performance after an ACL reconstruction. AB - The purpose of this study was to evaluate the validity of the simple measurements of the muscle performance of the lower extremity, one legged hop testing for distance and the measurement of the circumference of the thigh 15 cm proximal to the joint line, by comparing them to the isokinetic strength testing 5 to 9 years after an ACL reconstruction with a bone-patellar tendon-bone (BTB) autograft. The measurements were performed on 86 patients on average 7 years after the surgery. The clinical evaluation was performed using the standard knee ligament evaluation form of the International Knee Documentation Committee (IKDC) and the Lysholm and the Marshall knee scores. At seven years, the isokinetic mean strength deficit of knee extension at the 60 degrees per second was 10 % in the operated limb as compared to the contralateral limb (NS). In the knee flexion, the differences were even smaller and statistically not significant either. The strength deficit of the knee extension at all knee angle velocities (p < 0.005), and flexion at 60 degrees per second (p < 0.05), correlated to the one legged hop testing, so that the patients with the greatest strength deficit also had the worst outcome in the one legged hop test. Also, the correlation between thigh atrophy and the deficit in the isokinetic strength test was significant in knee extension at all knee angle velocities (p < 0.001), and in knee flexion at 180 degrees per second (p < 0.005). In the final evaluation of the IKDC, 21 patients were rated as "normal", 51 as "nearly normal", 13 as "abnormal", and one as "severely abnormal". The mean of the Lysholm score was 83, classified as good, and that of Marshall score 43, classified as good. In conclusion, the one legged hop testing, as a functional muscle strength testing after an anterior cruciate ligament reconstruction, seems to correlate well with the isokinetic strength testing of the knee, especially in the knee extension. Because the hop testing can be easily performed and without extra equipment, we recommend its use for the evaluation of the functional muscle performance after an anterior cruciate ligament reconstruction. The measurement of the thigh atrophy is also easy to perform, and should be used beside the one legged hop testing, especially if the isokinetic strength testing is not available. PMID- 11914984 TI - Ultrasound-guided percutaneous biopsies of the semitendinosus muscle following ACL reconstruction--a methodological description. AB - PURPOSE: To evaluate whether the ultrasound-guided biopsy technique could be used to analyse the histochemical profile of m. semitendinosus following ACL reconstruction with autologous semitendinosus tendon graft. MATERIAL AND METHODS: Sixteen patients were included in the study. During ACL surgery, specimens from m. semitendinosus were obtained via open biopsy. Ultrasound-guided biopsies from m. semitendinosus were performed 7 - 11 months postoperatively from operated leg (n = 16) and non-operated leg (n = 8). RESULTS: Two biopsy samples were pooled and formed a specimen of approximately 150 fibres. For fibre type composition, there was correlation between open biopsy and ultrasound-guided biopsy data, (P = 0.003, r = 0.89). The r-value for mean fibre area was 0.72 (P = 0.06). The methodological error for fibre type 1 % and mean fibre area was 9 %. Postoperatively, no significant differences were found in fibre type composition or fibre type areas between the operated and non-operated leg. CONCLUSION: Ultrasound-guided biopsy is an easy and safe technique that can be used for scientific studies and in clinical practice where biopsy from small muscles or from special location in muscles, such as in ruptures, scars or others is needed. PMID- 11914985 TI - Hyperthermia at 434 MHz in the treatment of overuse sport tendinopathies: a randomised controlled clinical trial. AB - The objective of this randomized study was to compare a thermotherapy system, hyperthermia at 434 MHz and conventional ultrasound in the treatment of overuse sports tendinopathies. The study group consisted of 44 athletes, 33 males and 11 females (age 26 +/- 4.56 years) affected by tendinopathies at lower extremities (patellar or achilles tendons). After elucidation of the kind of trial, 22 patients were randomly assigned to hyperthermia and 22 to ultrasound. The patients received after a pain measurement and ultrasound scanning 12 treatments, 3 times a week for 4 weeks. The same standardized examination was done at the end of treatment and 1 month after the end of treatment. The assessor physician was unaware of the treatment allocation. The patients were asked to rate the ultimate outcome on the base of pain resolution and return to sports activity. Both groups had a significant decrease of symptoms (P<0.001). Hyperthermia, however, demonstrated better effects on the reduction of VAS score and on the subjective overall satisfaction (77 %) of excellent and good results in comparison to the 33 % of ultrasound. In patients with chronic overuse tendinopathies hyperthermia at 434 MHz showed encouraging results, with short-term clinical improvement, safety and no side effects. PMID- 11914986 TI - Work-related acute injuries from mandatory fitness training in the Swedish Police Force. AB - Acute injuries in the Swedish Police Force from on-duty fitness training were selected retrospectively from the Information System of Occupational Injuries (ISA) at the National Board of Occupational Safety and Health and, if having caused a sick-leave exceeding 2 weeks, to the Labour Market Insurance (AMF Insurance). The latter included injuries from 1995 only. During the seven-year period 1992 to 1998, 920 injuries (80 % in males) from fitness training involving police officers were reported to the ISA-register. The total incidence was 1.6 for policemen per 10 000 hours of exposure and 2.2 for policewomen, which is 1.4 times higher than in men. Around 50 % of the injuries occurred in team and contact sports, with a slightly higher percentage for males, 54 % versus 49 % in females. The percentage of injuries from self-defense training was twice as high as in women than in men, 29 % versus 15 %. In 1995, 42 of the 72 injuries in males and 6 of the 21 injuries in females caused more than 14 days of sick-leave and were announced to the Occupational No Fault Liability Insurance. The major part, 32 of 48 injuries, came from team or contact sports (mainly floorball and soccer). Six policemen incurred injuries that were classified with a degree of disability ranging from 2 to 5 %. The total cost for medical treatment and production loss for the 48 injuries was Euro 248 448 and 99 336, respectively. Team and contact sports accounted for 89 % of the costs and 77 % of the production loss through sick-leave. PMID- 11914987 TI - The efficacy of acetic acid for glycogen repletion in rat skeletal muscle after exercise. AB - We examined the effect of acetic acid, the main component of vinegar, on glycogen repletion by using swimming-exercised rats. Rats were trained for 7 days by swimming. After an overnight fast, they were subjected to a 2-hr swimming exercise. Immediately afterward, they were given by gavage 2 ml of one of the following solutions: 30 % glucose only or 30 % glucose with 0.4 % acetic acid. Rats were sacrificed by decapitation before, immediately after exercise and 2 hours after the feeding. Exercise significantly decreased soleus and gastrocnemius glycogen content, and feeding significantly increased liver, soleus and gastrocnemius glycogen content. In soleus muscle, acetate feeding significantly increased glycogen content and the ratio of glycogen synthase in the I form (means +/- SEM: 4.04 +/- 0.41 mg/g-tissue and 47.0 +/- 0.7 %, respectively) in contrast to no acetate feeding (3.04 +/- 0.29 mg/g-tissue and 38.1 +/- 3.4 %, respectively). Thus, these findings suggest that the feeding of glucose with acetic acid can more speedily accelerate glycogen repletion in skeletal muscle than can glucose only. PMID- 11914988 TI - Naive and memory T cell subsets are differentially mobilized during physical stress. AB - This study examined the naive and memory phenotypic profiles of CD4+ and CD8hi T cells that were mobilized to the peripheral circulation during a combination of aerobic exercise and heat stress, determining expression of the adhesion molecules CD62L and CD11a on the recruited cells. Twelve recreationally active males (age 27.1 +/- 5.3 yr, height 1.77 +/- 0.08 m, mass 76.9 +/- 12.0 kg, VO2peak 43.9 +/- 6.7 mL x kg(-1) x min(-1)) completed a 40 min bout of cycle ergometry at 65 % of VO2peak while immersed to mid-chest in a water bath at 39 degrees C. Venous blood samples were collected before (T0), during (T40) and 30 min after (T70) exposure to combined exercise and heat stress. Specimens were analyzed by three-colour flow cytometry for CD4+ and CD8hi T cell expression of CD45RO, CD11a and CD62L. Some 80 % of the CD4+ T cells that were mobilized were of the CD45RO memory phenotype, with the numbers of CD11alo and CD62L+ cells increasing more than those of CD11ahi and CD62L- cells. For the CD8hi cells, there was a more equal recruitment of CD45RO- naive (43 %) and CD45RO+ memory (57 %) cells. The majority (84 %) of recruited CD8+ cells were CD11ahi; there was a trend to predominance of CD62L- cells (57 %) for the memory subset, but with almost equal recruitment of CD62L+/- for the naive subset. We conclude that the exercise + heat stress induced trend to an increase in CD4+ T cells is linked in some way to memory phenotype; it cannot be explained simply by a high density expression of CD11a and lack of the lymph node homing receptor (CD62L). Furthermore, although mobilization of CD8hi T cells is not linked to memory phenotype, a high density expression of CD11a and a lack of the lymph node homing receptor are important determinants of CD8hi T cell mobilization. PMID- 11914990 TI - Evidence for probable sexual transmission of the hepatitis g virus. AB - A cross-sectional epidemiology study evaluated the role of sexual activity and sexually transmitted diseases (STDs) in the transmission of hepatitis G virus (HGV/GBV-C) and other hepatitis virus infections in 944 subjects. There was a statistically significant higher prevalence of HGV/GBV-C, hepatitis B virus, and hepatitis C virus exposure in the STD clinic group (i.e., subjects who were currently seeking treatment for an STD) compared with the group who never had received treatment for an STD. In a comparison of the subjects with an STD versus those without an STD, the prevalence of HGV/GBV-C was 11.3% versus 4.9%, on the basis of polymerase chain reaction (PCR) results alone, and 36.6% versus 8.8%, when results of PCR and enzyme-linked immunosorbent assay were combined. Sexual activity and, possibly, the presence of an STD increases the risk of HGB/GBV-C transmission. PMID- 11914989 TI - Genomewide linkage analysis of quantitative spirometric phenotypes in severe early-onset chronic obstructive pulmonary disease. AB - Chronic obstructive pulmonary disease (COPD) is a common, complex disease associated with substantial morbidity and mortality. COPD is defined by irreversible airflow obstruction; airflow obstruction is typically determined by reductions in quantitative spirometric indices, including forced expiratory volume at 1 s (FEV(1)) and the ratio of FEV(1) to forced vital capacity (FVC). To identify genetic determinants of quantitative spirometric phenotypes, an autosomal 10-cM genomewide scan of short tandem repeat (STR) polymorphic markers was performed in 72 pedigrees (585 individuals) ascertained through probands with severe early-onset COPD. Multipoint variance-component linkage analysis (using SOLAR) was performed for quantitative phenotypes, including FEV(1), FVC, and FEV(1)/FVC. In the initial genomewide scan, significant evidence for linkage to FEV(1)/FVC was demonstrated on chromosome 2q (LOD score 4.12 at 222 cM). Suggestive evidence was found for linkage to FEV(1)/FVC on chromosomes 1 (LOD score 1.92 at 120 cM) and 17 (LOD score 2.03 at 67 cM) and to FVC on chromosome 1 (LOD score 2.05 at 13 cM). The highest LOD score for FEV(1) in the initial genomewide scan was 1.53, on chromosome 12, at 36 cM. After inclusion of 12 additional STR markers on chromosome 12p, which had been previously genotyped in this population, suggestive evidence for linkage of FEV(1) (LOD score 2.43 at 37 cM) to this region was demonstrated. These observations provide both significant evidence for an early-onset COPD-susceptibility locus on chromosome 2 and suggestive evidence for linkage of spirometry-related phenotypes to several other genomic regions. The significant linkage of FEV(1)/FVC to chromosome 2q could reflect one or more genes influencing the development of airflow obstruction or dysanapsis. PMID- 11914991 TI - A Cluster of cases of Mycobacterium szulgai keratitis that occurred after laser assisted in situ keratomileusis. AB - Laser-assisted in situ keratomileusis (LASIK) is a recently developed ophthalmic procedure. When 2 patients developed keratitis caused by Mycobacterium szulgai after they underwent LASIK surgery, we conducted a retrospective cohort study of all LASIK procedures performed at Scott & White Clinic (Temple, Texas) during a 4.5-month period. Seven patients had compatible symptoms and signs, 5 of whom had confirmed M. szulgai keratitis. Five cases occurred among 30 procedures performed by doctor A, and there were no cases among 62 procedures performed by doctor B (approximate relative risk, 12.0; 95% confidence interval, 1.6-679.0; P=.0029). Doctor A had chilled syringes of saline solution in ice for intraoperative lavage the only factor that differentiated the procedures of the 2 surgeons. Cultures of samples from the source ice machine's drain identified M. szulgai; the strain was identical to isolates recovered from all confirmed cases and differed from 4 standard M. szulgai strains, as determined by pulsed-field gel electrophoresis. Intraoperative contamination from ice water apparently led to M. szulgai keratitis in these patients. PMID- 11914992 TI - Pseudomonas aeruginosa ventilator-associated pneumonia: comparison of episodes due to piperacillin-resistant versus piperacillin-susceptible organisms. AB - We sought to determine the epidemiological characteristics of patients in an intensive care unit (ICU) who developed ventilator-associated pneumonia (VAP) caused by piperacillin-resistant Pseudomonas aeruginosa (PRPA; n=34) or piperacillin-susceptible P. aeruginosa (PSPA; n=101). According to univariate analysis, the factors associated with the development of PRPA VAP were presence of an underlying fatal medical condition, immunocompromised status, longer previous hospital stay, less-severe illness at the time of ICU admission, duration of mechanical ventilation before onset of VAP, number of classes of antibiotic received, and previous exposure to imipenem or fluoroquinolone. Multivariate logistic regression analysis identified the following significant independent factors: presence of an underlying fatal medical condition (odds ratio [OR], 5.6), previous fluoroquinolone use (OR, 4.6), and initial disease severity (OR, 0.8). We concluded that the clinical characteristics of patients who develop PRPA VAP differ from those of patients who develop PSPA VAP. Restricted fluoroquinolone use is the sole independent risk factor for PRPA VAP that is open to medical intervention. PMID- 11914993 TI - Antiviral susceptibility of Herpes simplex viruses and its clinical correlates: a single center's experience. AB - The in vitro susceptibility to acyclovir of 204 herpes simplex virus isolates from 165 immunocompromised patients treated at our hospital was determined by the cytopathic effect reduction assay. Approximately 95% of herpes simplex virus 1 and 73% of herpes simplex virus 2 isolates were inhibited by acyclovir at concentrations of <2 microgram/mL. From 8 patients (5%), an isolate with low susceptibility to acyclovir (50% inhibitory dose, >3 microgram/mL) was recovered. Medical records of 83 patients were reviewed. Lesions resolved in most of the patients, independent of treatment. Treatment failures were not always associated with isolation of an in vitro-resistant virus. On the contrary, when a virus with low susceptibility to acyclovir was isolated, resolution of the lesion was the rule. In 9 of 10 patients with subsequent recurrent episodes of disease, the susceptibility of the viruses isolated was similar to that of the first episode. Routine susceptibility testing in our geographic area is not encouraged because of the low incidence of acyclovir-resistant herpes simplex viruses. PMID- 11914994 TI - Prevalence and predictors of trimethoprim-sulfamethoxazole resistance among uropathogenic Escherichia coli isolates in Michigan. AB - Resistance among uropathogenic Escherichia coli to trimethoprim-sulfamethoxazole (TMP-SMX) has increased. Risk factors for resistance and the impact on clinical failure have been poorly described. We performed a retrospective cohort study of women with acute uncomplicated cystitis seen at a university health center and at primary care clinics in southeastern Michigan from 1992 to 1999. The prevalence of TMP-SMX resistance increased from 8.1% to 15.8% (P=.01). Women who had taken TMP-SMX recently were >16 times as likely as women who had not taken antibiotics recently to be infected with an isolate resistant to this agent; those who had taken any other antibiotic were more than twice as likely to be infected with a resistant isolate. Women infected with a TMP-SMX-resistant isolate who were treated with TMP-SMX were >17 times as likely to have treatment failure. Recent antibiotic use is a risk factor for infection with a TMP-SMX-resistant isolate; patients who are infected with a TMP-SMX-resistant isolate and who are treated with this agent are at a higher risk for clinical failure. PMID- 11914995 TI - Immunoglobulin a as a serological marker for the (silent) circulation of poliovirus in an inactivated poliovirus-vaccinated population. AB - Poliovirus-specific immunoglobulin A (IgA) is detected after infection with wild type virus or vaccination with live attenuated oral poliovirus (OPV) but not after vaccination with inactivated poliovirus (IPV). We examined whether the presence of IgA in serum can be used as a marker for poliovirus circulation in IPV-vaccinated populations in The Netherlands. In seronegative persons challenged with OPV, the sensitivity of this marker was 76%-86%. Results from a serosurvey showed a high seroprevalence (63%-73%) of IgA in the population born before vaccination was introduced in The Netherlands, which reflects natural exposure. The start of the vaccination program in 1957 corresponded to a reduction in the IgA seroprevalence in both vaccinated (2.1%-4.5%) and nonvaccinated groups (8.3% 11.7%). The presence of IgA-positive persons in the population could largely be explained by the occurrence of episodes of proven poliovirus circulation. We propose to use the detection of poliovirus-specific IgA as a tool to monitor virus circulation in IPV-vaccinated and nonvaccinated populations, to aid the poliovirus eradication process. PMID- 11914996 TI - A study evaluating the efficacy, safety, and tolerability of ertapenem versus ceftriaxone for the treatment of community-acquired pneumonia in adults. AB - In a double-blind, multicenter trial, 502 patients hospitalized with community acquired pneumonia were randomized to receive therapy with either ertapenem or ceftriaxone (for each, 1 g given intravenously once daily). After a minimum of 3 days, therapy could be switched to oral amoxicillin-clavulanate. The median duration of intravenously administered therapy for the 383 clinically evaluable patients was 4 days for both treatment groups; 345 patients (90.1%) had their treatment switched to orally administered therapy. Of the clinically evaluable patients, 168 (92.3%) in the ertapenem group and 183 (91.0%) in the ceftriaxone group had a favorable clinical response. Streptococcus pneumoniae was the most commonly isolated pathogen, and high cure rates were observed both for penicillin susceptible and -nonsusceptible infections in the ertapenem group (28 [87.5%] of 32 patients versus 17 [100%] of 17 patients, respectively). Both treatment regimens were generally well tolerated; the most common drug-related adverse events reported were diarrhea (2.9% versus 2.7%) and nausea (0.8% versus 2.0%) in the ertapenem and ceftriaxone groups, respectively. These results suggest that ertapenem and ceftriaxone therapy have similar efficacy and safety in hospitalized patients with community-acquired pneumonia. PMID- 11914997 TI - Clinical trials of immunomodulatory therapies in severe sepsis and septic shock. AB - Sepsis remains one of the leading causes of mortality in critically ill patients. Increased insight into the complexities of this disease process has resulted in the targeting of various aspects of the inflammatory response as offering potential therapeutic benefits. There have been encouraging results in the past few years. Some of the tested agents have been shown to improve mortality rates in large randomized controlled trials involving patients with severe sepsis. In this article, we discuss the positive and negative results of trials in this field; some of the possible reasons for the negative results are examined, and directions for the future are suggested. PMID- 11914998 TI - Definitions of cytomegalovirus infection and disease in transplant recipients. AB - Cytomegalovirus (CMV) infection and disease are important causes of morbidity and mortality among transplant recipients. For the purpose of developing consistent reporting of CMV in clinical trials, definitions of CMV infection and disease were developed and published. This study seeks to update the definitions of CMV on the basis of recent developments in diagnostic techniques, as well as to add to these definitions the concept of indirect effects caused by CMV. PMID- 11914999 TI - Opportunistic infections in patients with and patients without Acquired Immunodeficiency Syndrome. AB - In the next decade, longer survival of patients with cancer and more-aggressive therapies applied to common conditions, such as asthma and rheumatoid arthritis, will result in a larger population with significant immune system defects. Many in this population will be at risk for opportunistic infections, which are familiar to doctors who have treated people infected with human immunodeficiency virus (HIV). However, the epidemiology, presentation, and outcome of these infections in patients with an immune system defect, other than that caused by HIV infection, may be different than those encountered in patients with acquired immunodeficiency syndrome. Reviewed are 4 common opportunistic infections: Pneumocystis carinii pneumonia, cryptococcosis, atypical mycobacterial infection, and cytomegalovirus infection. Emphasized are the important differences among these groups at risk. PMID- 11915000 TI - Human immunodeficiency virus superinfection and recombination: current state of knowledge and potential clinical consequences. AB - Superinfection with multiple strains or subtypes of the human and simian immunodeficiency viruses has been documented. Recent increases in the prevalences of both unprotected anal intercourse and sexually transmitted diseases among men who have sex with men indicate that these men continue to practice unsafe sex and, therefore, are at risk for superinfection with the human immunodeficiency virus (HIV). Recurrent exposure to HIV among seropositive individuals who engage in high-risk behaviors can have serious consequences, because superinfection is a necessary first step for viral recombination to occur. Recombination may produce more virulent viruses, drug-resistant viruses, or viruses with altered cell tropism. Additionally, recombinant viruses and superinfection can accelerate disease progression and increase the likelihood of sexual transmission by increasing virus load in the blood and genital tract. The extent of superinfection and recombination in persons living with HIV is unknown. The implications of HIV superinfection and the generation of recombinant viruses are discussed. PMID- 11915001 TI - The consistency of adherence to antiretroviral therapy predicts biologic outcomes for human immunodeficiency virus-infected persons in clinical trials. AB - We prospectively studied long-term antiretroviral adherence patterns and their impact on biologic outcomes for human immunodeficiency virus (HIV)-infected participants in 2 randomized, multicenter clinical trials. For the period from baseline to month 12 of the study, participants who reported adherence levels of 100%, 80%-99%, and 0%-79% had plasma HIV RNA levels that decreased by 2.77, 2.33, and 0.67 log(10) copies/mL, respectively (P<.001), whereas their CD4 counts increased by 179, 159, and 53 cells/mm(3), respectively (P<.001). Adherence predicted nondetectable HIV RNA levels (<50 copies/mL) at 12 months of follow-up (P<.001). The HIV RNA level was nondetectable in 72% of participants who reported 100% adherence at all 4 follow-up visits, compared with 66%, 41%, 35%, and 13% of participants who reported 100% adherence at 3, 2, 1, or 0 follow-up visits, respectively (P<.001). Nonwhite race was associated with poorer adherence (P<.001), and older age was associated with better adherence (P<.001). PMID- 11915003 TI - Association of adherence to Mycobacterium avium complex prophylaxis and antiretroviral therapy with clinical outcomes in Acquired Immunodeficiency Syndrome. AB - High-level adherence to regimens of combination antiretroviral therapy and prophylactic medications for opportunistic infections (OIs) is crucial to their success. However, little is known about the association between adherence to these life-sustaining therapies and such clinical outcomes as progression of acquired immune deficiency syndrome (AIDS) or development of OIs. We assessed adherence to regimens of antiretroviral and Mycobacterium avium complex (MAC) prophylactic medications in 643 patients enrolled in a trial of MAC prophylaxis. By week 56 of the study follow-up, 42% of the patients reported nonadherence to MAC prophylaxis, whereas one-quarter of the patients reported nonadherence to potent antiretroviral regimens. Nonadherence to both MAC prophylaxis and antiretroviral therapy was associated with higher human immunodeficiency virus (HIV) type 1 RNA levels and a significant increase in the risk of developing an AIDS-defining complication or death. Predictors of nonadherence are presented. These results underscore the clinical significance of adherence to HIV therapy and may be helpful in designing interventions to optimize the management of HIV by improving adherence. PMID- 11915002 TI - Indinavir-associated interstitial nephritis and urothelial inflammation: clinical and cytologic findings. AB - The objective of the present study was to characterize the genitourinary syndromes that accompany indinavir-associated pyuria. Of 23 indinavir-treated patients with persistent pyuria, 4 had isolated interstitial nephritis, 10 had both interstitial nephritis and urothelial inflammation, 7 had isolated urothelial inflammation, and 2 had pyuria with nonspecific urinary tract inflammation. A total of 21 patients had multinucleated histiocytes identified by cytologic testing of urine specimens. Urine abnormalities resolved in all 20 patients who stopped receiving indinavir therapy. Pyuria continued in the 3 patients who continued receiving indinavir. Six patients had elevated serum creatinine levels, which returned to baseline levels when indinavir was discontinued. In conclusion, indinavir-associated pyuria was frequently associated with evidence of interstitial nephritis and/or urothelial inflammation, multinucleated histiocytes were commonly present in urine specimens, and cessation of indinavir therapy was associated with prompt resolution of urine abnormalities. PMID- 11915004 TI - Abacavir hypersensitivity reaction. AB - A hypersensitivity reaction occurs in association with initiation of abacavir therapy as part of combination antiretroviral therapy in approximately 3.7% of patients. The reaction is possibly the result of a combination of altered drug metabolism and immune dysfunction, which is poorly understood. White patients appear to be at higher risk and patients of African descent at lower risk of abacavir hypersensitivity. Clinical management involves supportive measures and discontinuation of abacavir therapy. Rechallenge with abacavir in a hypersensitive patient should be avoided because it might precipitate a life threatening reaction. PMID- 11915005 TI - Absence of opioid withdrawal symptoms in patients receiving methadone and the protease inhibitor lopinavir-ritonavir. AB - A study was designed to determine the interactions, both clinical and pharmacokinetic, between methadone and lopinavir-ritonavir. Results demonstrated a 36% reduction in the methadone area under the plasma concentration-time curve after the introduction of lopinavir-ritonavir, with no coincident symptoms of opioid withdrawal and no requirement for methadone dose adjustment. PMID- 11915006 TI - Emergence of multiple drug-resistant human cytomegalovirus variants in 2 patients with human immunodeficiency virus infection unresponsive to highly active antiretroviral therapy. AB - In 2 patients infected with human immunodeficiency virus (HIV), highly active antiretroviral therapy was unable to suppress HIV replication as a result of the emergence of drug-resistant HIV variants. Human cytomegalovirus (HCMV) retinitis developed in both patients, and an unusually complex mixture of drug-resistant HCMV variants was detected in both patients. PMID- 11915007 TI - Changes in the plasma lipid profile as a potential predictor of clinical outcome in dengue hemorrhagic fever. AB - In 50 consecutive children admitted to the intensive care unit with the clinical diagnosis of dengue hemorrhagic fever (DHF)/dengue shock syndrome (grade III or IV), 20 patients with mild DHF (grade I or II), and 20 healthy control patients, the plasma lipid profile was measured. Levels of total plasma cholesterol, high density lipoprotein, and low-density lipoprotein were significantly decreased in patients with the severest cases, compared with patients with mild DHF and healthy controls. Changes in the plasma lipid profile differentiate between patients with different stages of DHF disease severity and could be used as a potential predictor for clinical outcome. PMID- 11915008 TI - Patients with suspected herpes simplex encephalitis: rethinking an initial negative polymerase chain reaction result. AB - A statewide encephalitis diagnostic project of the California State Department of Health Services found that herpes simplex virus 1 DNA may not be detectable by molecular methods early in the clinical course of herpes simplex encephalitis. PMID- 11915009 TI - The effects of statins on mortality rates among bacteremic patients. PMID- 11915011 TI - Contamination of hospital water with Aspergillus fumigatus and other molds. PMID- 11915012 TI - Caspofungin in combination with itraconazole for the treatment of invasive aspergillosis in humans. PMID- 11915013 TI - Toxoplasmosis-associated deaths among Human Immunodeficiency Virus-infected persons in the United States, 1992-1998. PMID- 11915014 TI - Shoulder Girdle Syndrome revealing primary HIV infection. PMID- 11915015 TI - Prevention of travel-associated tuberculosis in the high-risk traveler. PMID- 11915016 TI - Drug-resistant Streptococcus pneumoniae in a day care population. PMID- 11915018 TI - Liver has rhythm. PMID- 11915019 TI - Nonalcoholic fatty liver disease: an agenda for clinical research. PMID- 11915020 TI - Obesity-related fatty liver is unchanged in mice deficient for mitochondrial uncoupling protein 2. AB - Nonalcoholic fatty liver disease (NAFLD), a prevalent condition associated with obesity, has the potential of evolving into end-stage liver disease. The biochemical mechanisms that define the progression of NAFLD are not well known, but reactive oxygen species (ROS) have been implicated in this process. Uncoupling protein (UCP) 2 is a mitochondrial inner-membrane protein that mediates proton leak, uncouples adenosine triphosphate (ATP) synthesis, and negatively regulates ROS production. UCP2 expression is increased in various animal models of NAFLD. Up-regulation of UCP2 may compromise cellular ATP levels and worsen liver damage, or it may be protective by ROS reduction in NAFLD. This study aimed to obtain a definitive answer as to whether increased UCP2 expression contributes to NAFLD. UCP2-/- mice were exposed to obesity by crossbreeding with ob/ob mice and by long-term high-fat feeding to study the effect of UCP2 deficiency on the outcome of NAFLD. Steatohepatitis score of crossbred mice (ob/ob/ko) was similar to that of ob/ob mice at 25 weeks. No compensatory increase was observed in the expression of UCP5 in ob/ob/ko livers. To unmask the effects of absent leptin and its potential proinflammatory actions, steatosis was also induced in UCP2-/- mice by a high-fat diet continued for 6 months. Serum alanine aminotransferase (ALT) levels remained normal, and the steatohepatitis score in UCP2-/- mice was the same as in wild-type controls. We conclude that increased expression of UCP2 in the livers of mice with genetically or diet induced obesity exerts neither protective nor deleterious effects on the severity of fatty liver disease. PMID- 11915022 TI - NF-kappaB inhibition sensitizes hepatocytes to TNF-induced apoptosis through a sustained activation of JNK and c-Jun. AB - Hepatocyte resistance to tumor necrosis factor alpha (TNF)-induced apoptosis is dependent on activation of the transcription factor nuclear factor kappaB (NF kappaB). To determine the mechanism by which NF-kappaB protects against TNF toxicity, the effect of NF-kappaB inactivation on the proapoptotic c-Jun NH(2) terminal kinase (JNK) signaling pathway was examined in the rat hepatocyte cell line RALA255-10G. Adenovirus-mediated NF-kappaB inactivation led to a prolonged activation of JNK and increased activating protein-1 (AP-1) transcriptional activity in response to TNF treatment. Inhibition of the function of the JNK substrate and AP-1 subunit c-Jun blocked cell death from NF-kappaB inactivation and TNF as determined by measures of cell survival, numbers of apoptotic and necrotic cells, and DNA hypoploidy. Inhibition of c-Jun function blocked mitochondrial cytochrome c release and activation of caspase-3 and -7. NF-kappaB therefore blocks the TNF death pathway through down-regulation of JNK and c Jun/AP-1. In conclusion, sustained JNK activation that occurs in the absence of NF-kappaB initiates apoptosis through a c-Jun-dependent induction of the mitochondrial death pathway. PMID- 11915021 TI - Leptin in hepatic fibrosis: evidence for increased collagen production in stellate cells and lean littermates of ob/ob mice. AB - Leptin is a 16-kd hormone that mediates a range of metabolic effects by using a transduction pathway from the long form of the leptin receptor, OB-R(L,) through Janus kinase-signal transducer and activator of transcription (Jak-Stat) signaling components. Leptin is produced by hepatic stellate cells (HSCs) but only following their "activation." Because activation of stellate cells is a central event in the fibrotic response to liver injury, we hypothesized that leptin may directly stimulate fibrogenesis in activated stellate cells via OB R(L). We analyzed leptin receptors and their signaling partners in a stellate cell line (HSC-T6) as well as in primary stellate cell isolates. We also examined the effect of leptin on stellate cell expression of alpha(2)(I) collagen messenger RNA (mRNA) levels by ribonuclease protection analysis (RPA). Finally, we examined the role of leptin in in vivo fibrogenesis by inducing a wounding response in ob/ob mice, which lack functional leptin. HSC-T6 and culture activated stellate cells expressed OB-R(L). Scatchard analysis verified specific binding of leptin to HSCs, with an association constant (K(d)) equal to 660 +/- 5.8 pmol/L. Exposure of HSCs to leptin resulted in significant increases in alpha(2)(I) collagen mRNA expression. Transient transfection with a promoter reporter construct showed a 3-fold increase in alpha(2)(I) collagen transgene activity. Leptin stimulated activation of Stat3 in activated HSCs. Finally, lean animals, but not ob/ob littermates, had significant fibrosis as assessed by picrosirius red staining and abundant alpha-smooth muscle actin staining. In conclusion, these results indicate that leptin is profibrogenic in activated HSCs and can signal via the Jak-Stat pathway. Up-regulation of leptin signaling in liver injury could contribute to enhanced fibrogenesis, particularly in states in which leptin levels are high. PMID- 11915023 TI - Inhibition of the MAPK and PI3K pathways enhances UDCA-induced apoptosis in primary rodent hepatocytes. AB - The mechanisms by which bile acids induce apoptosis in hepatocytes and the signaling pathways involved in the control of cell death are not understood fully. Here, we examined the impact of mitogen-activated protein kinase (MAPK) and phosphatidyl inositol 3-kinase (PI3K) signaling on the survival of primary hepatocytes exposed to bile acids. Treatment of hepatocytes with deoxycholic acid (DCA), chenodeoxycholic acid (CDCA) or ursodeoxycholic acid (UDCA) caused sustained MAPK activation that was dependent on activation of the epidermal growth factor receptor (EGFR). Activation of MAPK was partially blocked by inhibitors of PI3K. Inhibition of DCA-, CDCA-, and UDCA-stimulated MAPK activation resulted in approximately 20%, approximately 35%, and approximately 55% apoptosis, respectively. The potentiation of DCA- and CDCA-induced apoptosis by MEK1/2 inhibitors correlated with cleavage of procaspase 3, which was blocked by inhibitors of caspase 8 (ile-Glu-Thr-Asp-p-nitroanilide [IETD]) and caspase 3 (DEVD). In contrast, the potentiation of UDCA-induced apoptosis weakly correlated with procaspase 3 cleavage, yet this effect was also blocked by IETD and DEVD. Incubation of hepatocytes with the serine protease inhibitor AEBSF reduced the death response of cells treated with UDCA and MEK1/2 inhibitor to that observed for DCA and MEK1/2 inhibitor. The apoptotic response was FAS receptor- and neutral sphingomyelinase-dependent and independent of FAS ligand expression, and neither chelation of intracellular and extracellular Ca(2+) nor down-regulation of PKC expression altered the apoptotic effects of bile acids. In conclusion, bile acid apoptosis is dependent on the production of ceramide and is counteracted by activation of the MAPK and PI3K pathways. PMID- 11915024 TI - Maintaining HNF6 expression prevents AdHNF3beta-mediated decrease in hepatic levels of Glut-2 and glycogen. AB - The hepatocyte nuclear factor 3 (HNF-3) proteins are members of the Forkhead Box (Fox) family of transcription factors that play important roles in regulating expression of genes involved in cellular proliferation, differentiation, and metabolic homeostasis. In previous studies we increased liver expression of HNF 3beta by using either transgenic mice (transthyretin HNF-3beta) or recombinant adenovirus infection (AdHNF3beta), and observed diminished hepatic levels of glycogen, and glucose transporter 2 (Glut-2), as well as the HNF-6, HNF-3, HNF 1alpha, HNF-4alpha, and C/EBPalpha transcription factors. We conducted the present study to determine whether maintaining HNF-6 protein expression during AdHNF3beta infection prevents reduction of hepatic levels of glycogen and the earlier-mentioned genes. Here, we show that AdHNF3beta- and AdHNF6-infected mouse liver displayed increased hepatic levels of glycogen, Glut-2, HNF-3gamma, HNF 1alpha, and HNF-4alpha at 2 and 3 days postinfection (PI). Furthermore, restoration of hepatic glycogen levels after AdHNF3beta and AdHNF6 coinfection was associated with increased Glut-2 expression. AdHNF6 infection alone caused a 2-fold increase in hepatic Glut-2 levels, suggesting that HNF 6 stimulates in vivo transcription of the Glut-2 gene. DNA binding assays showed that only recombinant HNF-6 protein, but not the HNF-3 proteins, binds to the mouse -185 to -144 bp Glut-2 promoter sequences. Cotransfection assays in human hepatoma (HepG2) cells with either HNF-3 or HNF-6 expression vectors show that only HNF-6 provided significant transcriptional activation of the Glut-2 promoter. In conclusion, these studies show that the hepatic Glut-2 promoter is a direct target for HNF-6 transcriptional activation. PMID- 11915025 TI - Bone marrow transplantation in mice leads to a minor population of hepatocytes that can be selectively amplified in vivo. AB - Cell-based therapy may some day be a therapeutic alternative to liver transplantation. Recent observations indicating that hematopoietic stem cells can differentiate into hepatocytes have opened new therapeutic prospects. However, the clinical relevance of this phenomenon is unknown. We have previously developed a strategy based on the protective effect of Bcl-2 against Fas-mediated apoptosis to selectively amplify a small number of hepatocytes in vivo. We now show that this approach can be used to amplify a minor population of bone marrow derived hepatocytes. Normal mice were transplanted with unfractionated bone marrow cells from transgenic animals expressing Bcl-2 under the control of a liver-specific promoter. Recipients were then submitted to weekly injections of the anti-Fas antibody, Jo2. Upon sacrifice, the liver of the recipients showed bone marrow-derived clusters of mature hepatocytes expressing Bcl-2, which showed that the hepatocyte progeny of a genetically modified bone marrow can be selectively expanded in vivo. In contrast, no Bcl-2 expression could be detected without the selective pressure of Jo2, suggesting that differentiation of bone marrow cells into mature hepatocytes is very inefficient under physiologic conditions. We conclude that a selection strategy will be required to achieve a therapeutic level of liver repopulation with bone marrow-derived hepatocytes. PMID- 11915026 TI - The resistance of P. acnes--primed interferon gamma-deficient mice to low-dose lipopolysaccharide-induced acute liver injury. AB - Endotoxin has been identified as a principal mediator of sepsis, often with resulting multiple organ failure. Although interferon gamma (IFN-gamma) has a central role in controlling bacterial infection through the activation of macrophages and T lymphocytes, it can also enhance the harmful effects of the inflammatory response. To examine the role of IFN-gamma in lipopolysaccharide (LPS)-induced injury, we administered LPS (20 or 800 microg/mouse) alone or as low-dose LPS (20 microg/mouse) 7 days after heat-killed Propionibacterium acnes (P. acnes) injection into wild-type C57BL/6 (B6) mice or IFN-gamma-deficient (GKO) mice (B6 background). Although low-dose (20 microg) LPS alone had no effect on survival, the administration of 800 microg LPS alone resulted in 100% mortality in both B6 and GKO mice without significant hepatic mononuclear cellular infiltration or differences in elevated plasma tumor necrosis factor alpha (TNF-alpha), interleukin 6 (IL-6), and IL-12 levels. In contrast, mortality after low-dose (20 microg) LPS challenge in P. acnes-primed B6 mice was 100%, but 0% in GKO mice. In vivo plasma cytokine (IFN-gamma, TNF-alpha, IL-6, and IL-12) levels and in vitro cytokine production by hepatic mononuclear cells were significantly higher in B6 mice compared with GKO mice. Associated hepatic mononuclear cellular infiltration, multifocal liver necrosis, hepatomegaly, and splenomegaly were found in B6 mice, but not in GKO mice. Finally, the anti inflammatory NK1.1+CD4+ cell proportion of hepatic infiltrating mononuclear cell numbers 7 days after P. acnes administration was significantly reduced in B6 compared with GKO mice, whereas the proportion of inflammatory NK1.1+CD4- cells was increased. In conclusion, these data suggest that IFN-gamma mediates P. acnes primed low-dose LPS injury through the hepatic infiltration of mononuclear cells and the subsequent elevation of inflammatory cytokines after LPS challenge, whereas the lethal effects of high-dose LPS alone does not depend on the presence of IFN-gamma. PMID- 11915027 TI - Ex vivo exposure to carbon monoxide prevents hepatic ischemia/reperfusion injury through p38 MAP kinase pathway. AB - A direct role of carbon monoxide (CO), an effector-signaling molecule during heme oxygenase-1 (HO-1) catalysis of heme, in the protection against hepatic ischemia/reperfusion (I/R) injury needs to be established. This study was designed to determine the effects and downstream mechanisms of CO on cold I/R injury in a clinically relevant isolated perfusion rat liver model. After 24 hours of cold storage, rat livers perfused ex vivo for 2 hours with blood supplemented with CO (300 parts per million) showed significantly decreased portal venous resistance and increased bile production, as compared with control livers perfused with blood devoid of CO. These beneficial effects correlated with improved liver function (serum glutamic oxaloacetic transaminase levels) and diminished histological features of hepatocyte injury (Banff's scores). The CO mediated cytoprotective effects were nitric oxide synthase- and cyclic guanine monophosphate-independent, but p38 mitogen-activated protein kinase (MAPK) dependent. Moreover, adjunctive use of zinc protoporphyrin, a competitive HO-1 inhibitor, has shown that exogenous CO could fully substitute for endogenous HO-1 in preventing hepatic I/R insult. This study performed in a clinically relevant ex vivo cold ischemia model is the first to provide the evidence that HO-1 mediated cytoprotection against hepatic I/R injury depends on the generation of, and can be substituted by, exogenous CO. The p38 MAPK signaling pathway represents the key downstream mechanism by which CO prevents the I/R insult. In conclusion, regimens that employ exogenous CO should be revisited, as they may have potential applications in preventing/mitigating I/R injury, and thus expanding the liver donor pool for clinical transplantation. PMID- 11915028 TI - The in vivo apoptotic effect of interferon alfa-2b on rat preneoplastic liver involves Bax protein. AB - To determine whether interferon alfa (IFN-alpha) prevents in vivo oncogenesis in very-early-stage cancer cells, we evaluated the action of IFN-alpha2b over preneoplastic foci in rats. Animals were divided into 6 groups: subjected to a 2 phase model (diethylnitrosamine [DEN] plus 2-acetylaminofluorene [2-AAF]) of preneoplasia development (group 1), treated with IFN-alpha2b during the 2 phases (group 2), only during initiation with DEN (group 3), only during administration of 2-AAF (group 4), subjected only to an initiation stage (group 5), and treated with IFN-alpha2b during this period (group 6). The numbers of placental form of rat glutathione S-transferase (rGST-P)-positive foci per liver and the foci as percentage of liver were significantly reduced in groups 2, 3, and 6 but not in group 4. Rats treated with IFN-alpha2b showed a higher apoptotic index (AI) in altered hepatic foci (AHF). Levels of p53 and Bax protein in liver lysates were significantly increased in those animals. Similarly, levels of antiapoptotic proteins Bcl-2 and Bcl-x(L) in mitochondrial fraction were decreased. Finally, increased levels of Bax protein were localized in the mitochondria of rats that received IFN-alpha2b, at least during the DEN phase (groups 2, 3, and 6), whereas mitochondrial Bax expression was not increased in group 4. In conclusion, the preneoplastic hepatocytes in rats that received IFN-alpha2b during the initiation stage undergo programmed cell death as a primary result of a significant increase in the amount and translocation to the mitochondria of Bax protein. PMID- 11915029 TI - Synergistic effect of basic fibroblast growth factor and vascular endothelial growth factor in murine hepatocellular carcinoma. AB - The growth of any solid tumor depends on angiogenesis. Among the known angiogenic factors, basic fibroblast growth factor (bFGF) and vascular endothelial growth factor (VEGF), are potent and representative factors involved in tumor development. It has been reported that bFGF and VEGF showed a synergistic effect in both in vitro and in vivo angiogenesis. However, the interaction of these factors on tumor development and angiogenesis, including hepatocellular carcinoma (HCC), has not yet been elucidated. In this study, we examined the combined effect of bFGF and VEGF overexpression by means of a combination of a retroviral tetracycline (tet)-regulated (Retro-Tet) gene expression system, which can manipulate the gene expression in vivo by providing tet in the drinking water, and a conventional plasmid gene expression system. In an allograft study, bFGF and VEGF overexpression synergistically increased tumor growth and angiogenesis in the murine HCC cells. This synergistic effect also was found in established tumors. VEGF messenger RNA (mRNA) expression in the tumor was increased 3.1-fold by bFGF-overexpression, and the bFGF-induced tumor development was significantly attenuated by treatment with KDR/Flk-1 neutralizing monoclonal antibody. In conclusion, these results suggest that bFGF synergistically augments VEGF mediated HCC development and angiogenesis at least partly by induction of VEGF through KDR/Flk-1. PMID- 11915030 TI - The MDR phenotype is associated with the expression of COX-2 and iNOS in a human hepatocellular carcinoma cell line. AB - The presence of multiple drug resistance (MDR1) and angiogenic phenotypes negatively affect patients' prognosis with cancer even when treated with drugs that are not transported by the MDR1 gene product. It is possible to suggest a link between the MDR1 and angiogenic phenotypes. Because prostaglandins (PGs) and nitric oxide (NO) have been proposed to be involved in angiogenesis in vivo, the production of PGs and NO and the behavior of inducible NO synthase (iNOS), cyclooxygenase 1 (COX-1), and inducible cyclooxygenase (COX-2) were studied in parental drug-sensitive (P5) liver cancer cell lines and in P5-derived MDR1 cells P1(0.5). Immunohistochemical evaluation, Northern and Western blot analysis of COX-2 and iNOS, and assessment of cell proliferation were performed in basal conditions and after the exposure to stimulants or to specific inhibitors of COX 2 and iNOS. The messenger RNA and protein levels of COX-2 and iNOS were in basal conditions higher in P1(0.5) cells than the parental P5 cells. The exposure to lipopolysaccharide (LPS) or epidermal growth factor (EGF) determined an increase of PG and NO production in both cell lines and this increase was strongly reduced by COX-2 inhibitors such as celecoxib (CLX) and nimesulide (NIME). The inhibition of NO production by COX-2 inhibitors suggests cross-talk between COX-2 and iNOS pathways. CLX and NIME also inhibited cell proliferation, but only in MDR1 cells. A specific inhibitor of iNOS, N(6)-(1-iminoethyl)-L-lysine, had only a mild effect on cell proliferation in both cell lines. In conclusion, these data support the hypothesis that the MDR1 and angiogenic phenotypes are linked to each other in human liver cancer cell lines. PMID- 11915031 TI - alpha-Fetoprotein mRNA in the circulation as a predictor of postsurgical recurrence of hepatocellular carcinoma: a prospective study. AB - alpha-fetoprotein (AFP) messenger RNA (mRNA) has been proposed as a marker of hepatocellular carcinoma (HCC) cells disseminated into the circulation, but its clinical significance remains controversial. We prospectively assessed the prognostic value of AFP mRNA in patients undergoing curative hepatic resection for HCC. Peripheral blood samples were taken from 87 patients before and after surgery to determine the presence of AFP mRNA by use of a reverse-transcription polymerase chain reaction. A primary endpoint was recurrence-free interval. AFP mRNA was detectable preoperatively in 31 patients (36%) and postoperatively in 30 patients (34%). With a median follow-up period of 28 months (range, 3-41 months), HCC recurred in 46 patients (53%). Among 4 groups separated according to preoperative and postoperative AFP mRNA status, patients with consistent positivity of AFP mRNA showed the highest recurrence rate (85%) and trend to distant or multiple recurrence. The recurrence-free interval was significantly shorter in patients with postoperative positivity of AFP mRNA than in those without (53% [95% CI, 36-71] vs. 88% [95% CI, 79-96] at 1 year, 37% [95% CI, 17 57] vs. 60% [95% CI, 46-75] at 2 years; P =.014), whereas the preoperative positivity of AFP mRNA provided no significance (P =.100). Cox's proportional hazards model identified the postoperative positivity of AFP mRNA as an independent prognostic factor for HCC recurrence (relative risk, 2.33; 95% CI, 1.26-4.34; P =.007). In conclusion, postsurgical recurrence of HCC can be predicted by detecting AFP mRNA-expressing cells in peripheral blood. PMID- 11915032 TI - Tie2 vascular endothelial receptor expression and function in hepatocellular carcinoma. AB - Hepatocellular carcinoma (HCC) is generally characterized as a hypervascular tumor of rapid growth. We have previously reported that angiopoietin (Ang), a ligand for Tie2 vascular endothelial-specific receptor tyrosine kinase, may play a role in the progression of human HCC (J Clin Invest 1999;103:341-345) and matrix proteinase expression (Cancer Res 2001;61:2145-2153). However, the role of Tie2 receptor in hepatic oncogenesis is unknown. The Tie2 receptor protein was overexpressed in the neovascular endothelium of 31 of 39 (80%) human HCC tumors by immunohistochemical analysis with significant correlation to cell dedifferentiation and tumor size (P <.05). In vitro expression of a dominant negative construct, containing a soluble Tie2 ectodomain (sTie2), led to Ang protein interaction, inhibition of endogenous Tie2 phosphorylation in vascular endothelial cells and matrix metalloproteinase 9 (MMP-9) suppression. In conclusion, tumorigenicity with neovascularization was suppressed by in vivo gene transfer and sTie2 expression in a murine HCC model, suggesting a possible role for Tie2 expression in the induction of HCC neovascularization and disease progression. Inhibition of the Ang/Tie2 signal transduction cascade is a promising approach for tumor treatment. PMID- 11915033 TI - Lower risk for alcohol-induced cirrhosis in wine drinkers. AB - Although there is a well-known relationship between total alcohol intake and future risk for cirrhosis, other factors such as the type of alcohol consumed are sparsely studied. The aim of this study was to assess the effects of wine compared with other types of alcoholic beverages on risk for alcohol-induced cirrhosis. In 3 prospective studies, 30,630 participants from the Copenhagen area were followed-up for a total observation time of 417,325 person-years. Information on weekly intake of beer, wine, and spirits, and sex, age, body mass index, smoking habits, and education was obtained from questionnaires. The primary outcome measures were first admission or death, with alcohol-induced cirrhosis obtained from death certificates and from the National Hospital Discharge Register. Data were analyzed by means of multiplicative Poisson regression models. We confirmed the increasing risk for cirrhosis with increasing alcohol intake. Individuals who drank more than 5 drinks per day had a relative risk of 14 to 20 for developing cirrhosis compared with non- or light drinkers. However, compared with individuals who drank no wine (relative risk set at 1.0), individuals drinking 16% to 30% wine of their total intake had a relative risk of 0.4 (95% confidence limits, 0.3-0.6) and those drinking 51% or more of wine had a relative risk of 0.3 (95% confidence limits, 0.2-0.5) for developing cirrhosis. In conclusion, the results suggest that a high intake of all 3 types of alcohol conveys an increased risk for cirrhosis, but wine drinkers are at a lower risk than beer and spirits drinkers. PMID- 11915034 TI - Acute versus chronic alcohol consumption in acetaminophen-induced hepatotoxicity. AB - The aim of this study was to determine by multivariate analysis how alcohol and other factors affect the clinical course and outcome in patients with acetaminophen (paracetamol) poisoning. A total of 645 consecutive patients admitted from 1994 to 2000 with single-dose acetaminophen poisoning were studied, giving special attention to alcohol history, time between overdose and intravenous N-acetylcysteine (NAC) treatment ("time to NAC"), and other data available at the time of admittance. Up until 72 hours after ingestion, time to NAC was the single most important independent risk factor. With a time to NAC less than 12 hours, the mortality rate was 0.42% (95% CI, 0.05-2.7). When time to NAC exceeded 12, 24, and 48 hours, the mortality rate increased to 6.1%, 13%, and 19%, respectively. Chronic alcohol abuse was an independent risk factor of mortality (odds ratio [OR], 3.52; 95% CI, 1.78-6.97). Acute alcohol ingestion was an independent protective factor regarding mortality in alcoholic patients (OR, 0.08; 95% CI, 0.01-0.66) but not in nonalcoholic patients (OR, 0.21; 95% CI, 0.03 1.67). Patient age and quantity of acetaminophen were independent risk factors. In conclusion, time to NAC was confirmed as the major risk factor in acetaminophen-induced hepatotoxicity and mortality. Chronic alcohol abuse was an independent risk factor that could be counteracted by concomitant acute alcohol ingestion. We suggest that patients with chronic alcoholism and suspected acetaminophen poisoning due to an increased risk of developing hepatotoxicity should be treated with NAC regardless of risk estimation. PMID- 11915035 TI - Polymorphism of the N-acetyltransferase 2 gene as a susceptibility risk factor for antituberculosis drug-induced hepatitis. AB - Antituberculosis drug-induced hepatitis is one of the most prevalent drug-induced liver injuries. Isoniazid is the major drug incriminated in this hepatotoxicity. Isoniazid is mainly metabolized to hepatotoxic intermediates by N acetyltransferase (NAT). However, the association of polymorphic NAT acetylator status and antituberculosis drug-induced hepatitis is debatable. To determine whether acetylator status is a risk factor for antituberculosis drug-induced hepatitis, we genotyped NAT2 in 224 incident tuberculosis patients who received antituberculosis treatment. Antituberculosis drug-induced hepatitis was diagnosed based on a positive isoniazid rechallenge test and exclusion of viral hepatitis. Acetylator status was determined by genotyping NAT2 in patients using a polymerase chain reaction with restriction fragment length polymorphism. Univariate analysis and logistic regression analysis were used to evaluate the risk factors of isoniazid-induced hepatitis. Thirty-three patients (14.7%) were diagnosed with antituberculosis drug-induced hepatitis. Slow acetylators had a higher risk of hepatotoxicity than rapid acetylators (26.4% vs. 11.1%, P =.013). Among patients with hepatotoxicity, slow acetylators had significantly higher serum aminotransferase levels than rapid acetylators. Logistic regression showed that slow-acetylator status (odds ratio [OR], 3.66; 95% CI, 1.58-8.49; P =.003) and age (OR, 1.09; 95% CI, 1.04-1.14; P <.001) were the only 2 independent risk factors for antituberculosis drug-induced hepatitis. In conclusion, slow acetylator status of NAT2 is a significant susceptibility risk factor for antituberculosis drug-induced hepatitis. Additionally, slow acetylators are prone to develop more severe hepatotoxicity than rapid acetylators. Regular monitoring of serum aminotransferase levels is mandatory in patients receiving antituberculosis treatment, especially in slow acetylators. PMID- 11915036 TI - Sustained remission after corticosteroid therapy for type 1 autoimmune hepatitis: a retrospective analysis. AB - Autoimmune hepatitis commonly relapses after corticosteroid therapy, and long term management strategies have been proposed based on the premise that repeated relapses after drug withdrawal are inevitable. Our goal was to determine the frequency that remission can be sustained after its induction and termination of therapy. A total of 107 patients who had entered remission on conventional regimens were assessed for sustained remission after initial treatment and after relapse and re-treatment. Re-treatment strategies included conventional regimens and long-term maintenance schedules. Twenty-two patients (21%) achieved a sustained remission after initial treatment, and 24 of 85 patients who relapsed and were re-treated (28%) had a similar outcome. The probability of a sustained remission was 47% after 10 years of follow-up. Patients who sustained remission after initial therapy were distinguished only by a lower serum gamma-globulin level at entry. Conventional re-treatment schedules after relapse were able to induce a sustained remission more commonly then long-term maintenance schedules (59% vs. 12%, P =.00002). In conclusion, patients who respond to initial corticosteroid therapy can achieve a sustained remission after treatment withdrawal or after relapse and re-treatment. All patients are candidates for this outcome, and withdrawal of medication, even during maintenance schedules, is necessary to assess its likelihood. PMID- 11915037 TI - Apolipoprotein synthesis in nonalcoholic steatohepatitis. AB - The pathophysiology of hepatic steatosis, a prerequisite of nonalcoholic fatty liver disease, is poorly understood. Because very-low-density lipoprotein (VLDL) formation is the chief route of hepatic lipid export, we hypothesized that the synthesis of apoB-100, a rate-determining step in hepatic VLDL formation, may be altered in patients with nonalcoholic steatohepatitis (NASH). This study evaluated the relative synthesis rates of apolipoprotein B-100 (apoB-100) in patients with NASH and in lean and body mass index (BMI)-matched (obese) controls without NASH. A primed continuous infusion of L-[1-(13)C] leucine was used to measure the absolute synthesis rates (ASR) of apoB-100 and fibrinogen in 7 patients with NASH and compared them with 7 lean and 7 obese (BMI-matched) controls without NASH. The ASRs of fibrinogen and albumin also were measured. The mean ASR of apoB-100 in patients with NASH was lower (31.5 +/- 3.4 mg/kg/d) than that of obese (115.2 +/- 7.2 mg/kg/d, P <.001) and lean controls (82.4 +/- 4.1 mg/kg/d, P =.002). In contrast, the mean ASR of fibrinogen was greater in subjects with NASH than in both control groups. These data indicate that NASH is associated with markedly altered hepatic synthesis of apoB-100. The finding that albumin synthesis was not similarly decreased in patients with NASH shows that the attenuation of apoB-100 synthesis is not on the basis of globally impaired hepatic protein synthesis. In conclusion, because apoB-100 synthesis is a rate determining step in hepatocyte lipid export, decreased synthesis of this protein may be an important factor in the development of hepatic steatosis, a prerequisite for NASH. PMID- 11915038 TI - Hemolysis and bilirubin conjugation in association with UDP glucuronosyltransferase 1A1 promoter polymorphism. AB - Hemolysis may contribute to hyperbilirubinemia in Gilbert's syndrome. The authors examined blood carboxyhemoglobin corrected for inspired CO (COHbc) to index heme catabolism and serum conjugated bilirubin fractions to reflect bilirubin conjugation. Both parameters were related to UDP-glucuronosyltransferase 1A1 (UGT) promoter polymorphism, associated with Gilbert's syndrome, in term male newborns. COHbc was expressed as percentage of total hemoglobin, and total conjugated bilirubin (TCB) value as a percentage of serum total bilirubin (STB), (TCB/STB[%]). A production/conjugation index, COHbc/(TCB/STB[%]), represented bilirubin production divided by conjugation. UGT promoter genotype was designated according to the number of promoter TA insertions in each allele: 6/6, homozygous normal; 6/7, heterozygous; 7/7, homozygous variant. STB and COHbc values were higher in the 7/7 subgroup than the other counterparts (P <.01). The COHbc/(TCB/STB[%]) was higher in the 7/7 than either the 6/6 or 6/7 subsets (1.93 [1.31-2.88] vs. 0.85 [0.51-1.72] and 0.84 [0.53-1.87], respectively; P <.01). In conclusion, 7/7 UGT promoter polymorphism was associated with increased blood COHbc values (unexpected finding) as well as diminished serum total conjugated bilirubin ratios (expected finding). The increased hemolysis may contribute to the pathogenesis of increased STB values seen in Gilbert's syndrome, and exacerbate neonatal hyperbilirubinemia associated with the promoter polymorphism. PMID- 11915039 TI - Uroporphyria in mice: thresholds for hepatic CYP1A2 and iron. AB - In mice treated with 5-aminolevulinic acid (ALA) and polyhalogenated aromatic compounds, the levels of both hepatic cytochrome P450 (CYP)1A2 and iron-which can be quite different among inbred strains-are critical in causing experimental uroporphyria. Here we investigate the development of uroporphyria as a function of CYP1A2 and iron levels in the liver of mice having a common C57BL/6 genetic background. We compared Cyp1a2(-/-) knockout mice, Cyp1a2(+/-) heterozygotes, Cyp1a2(+/+) wild type, and Cyp1a2(+/+) mice pretreated with a low dose of 3,3',4,4',5-pentachlorobiphenyl (PCB126) (4 microg/kg). Cyp1a2(+/-) mice contain about 60% of the hepatic CYP1A2 content of Cyp1a2(+/+) mice, and the PCB126 pretreated Cyp1a2(+/+) mice have about twice the wild-type levels of CYP1A2. ALA- and iron-treated Cyp1a2(+/+) mice are known to accumulate hepatic uroporphyrin; this accumulation was increased 7-fold by pretreatment with the low dose of PCB126. ALA- and iron-treated Cyp1a2(+/-) heterozygote mice accumulated no uroporphyrin in 4 weeks, but by 8 weeks accumulated significant amounts of uroporphyrin. As previously reported, the ALA- and iron-treated Cyp1a2(-/-) knockout mouse has no CYP1A2 and exhibits no detectable uroporphyrin accumulation. Iron dose-response curves in ALA- and PCB126-treated Cyp1a2(+/+) mice showed that hepatic iron levels greater than 850 microg/g liver were required to produce significant uroporphyrin accumulation in the liver. Other measures of hepatic effects of iron (iron-response element-binding protein [IRP] iron response element [IRE] binding activity and accumulation of protoporphyrin from ALA) decreased when the level of iron was considerably lower than 850 microg/g liver. At low iron doses, accumulation of iron was principally in Kupffer cells, whereas at the higher doses (required to stimulate uroporphyrin accumulation), more iron was found in parenchymal cells. We conclude that small changes in hepatic CYP1A2 levels can dramatically affect uroporphyria in C57BL/6 mice, providing the animals have been sufficiently loaded with iron; these data might be clinically relevant to acquired (sporadic) porphyria cutanea tarda, because humans show greater than 60-fold genetic differences in hepatic basal CYP1A2. PMID- 11915040 TI - Hepatitis B e antigen in sera from individuals infected with hepatitis B virus of genotype G. AB - Hepatitis B virus (HBV) genotype G (HBV/G) was detected in sera from four individuals by polymerase chain reaction with hemi-nested primers deduced from an insertion of 36 nt in the core gene that is specific for this genotype. Despite two stop codons in the precore region characteristic of HBV/G, all patients were positive for hepatitis B e antigen (HBeAg) in serum. When 10 HBV clones were propagated from one patient, and sequenced within precore region and a section of the core gene, 6 clones were HBV/G while 2 were genotype A (HBV/A); a recombination between HBV/G and HBV/A occurred in the remaining 2 clones. Mixed infection of HBV/G and HBV/A, as well as the recombination, was demonstrated in the sequence of preS1 and preS2 regions also. Coinfection with HBV/G and HBV/A was demonstrated in the other three patients, and their recombination in two patients. Ten HBV clones were propagated from one patient at two time points separated by 1 year. Clones of HBV/A, HBV/G and their recombination were found in 9 : 1 : 0 when the patient was positive for HBeAg, while the proportion shifted to 0 : 8 : 2 after the patient seroconverted to anti-HBe. In conclusion, HBV/G is frequently found as a coinfection with HBV/A. This coinfection would explain the presence of HBeAg in individuals infected with HBV/G. Along with seroconversion to anti-HBe, HBV/G would be selected accompanied by the recombination with HBV/A. Further studies should be performed to confirm these findings. PMID- 11915042 TI - Interaction of hepatitis C virus core protein with retinoid X receptor alpha modulates its transcriptional activity. AB - Hepatic steatosis and hepatocellular carcinoma (HCC) are common and serious features of hepatitis C virus (HCV) infection, and the core protein has been shown to play distinct roles in the pathogenesis. Here we report the direct interaction of HCV core protein with retinoid X receptor alpha (RXRalpha), a transcriptional regulator that controls many aspects of cell proliferation, differentiation, and lipid metabolism. The core protein binds to the DNA-binding domain of RXRalpha, leading to increase the DNA binding of RXRalpha to its responsive element. In addition, RXRalpha is activated in cells expressing the core protein as well as in the livers of the core-transgenic mice that would develop hepatic steatosis and HCC later in their lives. Using promoter genes of cellular retinol binding protein II (CRBPII) and acyl-CoA oxidase as reporters, we also show that the expression of the core protein enhances the transcriptional activity regulated by the RXRalpha homodimer as well as by the heterodimer with peroxisome proliferator activated receptor alpha. Furthermore, expression of the CRBPII gene is also up-regulated in the livers of HCV core-transgenic mice. In conclusion, these results suggest that modulation of RXRalpha-controlled gene expression via interaction with the core protein contributes to the pathogenesis of HCV infection. PMID- 11915041 TI - Viral kinetics in genotype 1 chronic hepatitis C patients during therapy with 2 different doses of peginterferon alfa-2b plus ribavirin. AB - Pegylated interferon (peginterferon) alfa-2b plus ribavirin achieves a higher sustained response rate in patients with genotype 1 chronic hepatitis C virus (HCV) than standard combination therapy. This study evaluated HCV kinetics throughout therapy with 2 doses of peginterferon alfa-2b and ribavirin in 55 patients. Twenty-eight patients were randomized to receive a high once-weekly dose of peginterferon alfa-2b (3 microg/kg for 1 week, 1.5 microg/kg for 3 weeks, and 1.0 microg/kg for 44 weeks), and 27 patients were randomized to receive a low dose (0.5 microg/kg) for 48 weeks. Both groups also received 800 mg ribavirin daily. Mean baseline HCV RNA load, measured by reverse-transcription polymerase chain reaction, was similar in both groups (5.32 +/- 0.86 log vs. 5.15 +/- 1.04 log). The 3-microg/kg dose of peginterferon alfa-2b inhibited HCV RNA more significantly than the 0.5-microg/kg dose during the first 48 hours (2.08 +/- 0.93 log vs. 1.09 +/- 0.80 log; P <.001) and both increased at 72 hours (0.54 +/- 0.73 log vs. 0.03 +/- 0.36 log; P = not significant [NS]), but the high dose showed a greater reduction at the end of the week (1.07 +/- 0.99 log vs. 0.72 +/- 0.73 log). Both doses showed a progressive, slower viral decrease throughout therapy; however, HCV RNA became undetectable faster and in more patients with the high dose (22% vs. 7% at week 4, 56% vs. 44% at week 12, 69% vs. 63% at week 24, and 71% vs. 61.5% at the end of therapy). In conclusion, peginterferon alfa 2b/ribavirin produces an initial rapid decline in HCV RNA levels, followed by a slower, progressive decrease, similar to the biphasic kinetic profile of standard combination therapy. Higher doses of peginterferon alfa-2b also accelerate viral clearance. PMID- 11915043 TI - Peripheral blood count abnormalities among patients with hepatitis C in the United States. AB - An estimated 2.7 million people in the United States are infected with the hepatitis C virus (HCV), yet the influence of HCV infection on the peripheral blood count remains unknown. To investigate the prevalence of low peripheral blood counts among HCV-infected adults in the United States general population, we analyzed data collected in the third National Health and Nutrition Examination Survey (NHANES III). The study population consisted of 16,196 individuals age 18 or older who had peripheral blood counts and data on HCV infection. The lowest fifth percentile of each component of the peripheral blood was designated a priori as being low. HCV infection was assessed by antibody reactivity. HCV antibody-positive individuals were 3-fold more likely to have low neutrophil counts (HCV positive, 9% vs. HCV negative, 3%, P <.0001) and 2.6-fold more likely to have low platelet counts (HCV positive, 13% vs. HCV negative, 5%, P <.0001) independent of other evaluated factors. HCV infection was observed in more than 20% of persons with neutrophil counts below 1.0 x 10(9)/L or platelet counts less than 100 x 10(9)/L. No association was detected between anti-HCV status and anemia or other peripheral blood cell components. In conclusion, HCV-infected persons in the general population of the United States are more likely to have low neutrophil and platelet counts, and HCV testing should be considered for persons with unexplained neutrophil counts below 1.0 x 10(9)/L or platelet counts less than 100 x 10(9)/L. Alternate causes of anemia should be considered for HCV infected persons with low red blood cell counts. PMID- 11915044 TI - SEN virus: response to interferon alfa and influence on the severity and treatment response of coexistent hepatitis C. AB - The SEN virus (SENV) is a recently identified single-stranded, circular DNA virus. A strong association between 2 SENV variants (SENV-D and SENV-H) and transfusion-associated non-A-to-E hepatitis has been reported. To clarify the effect of SENV infection on coexisting chronic hepatitis C and the effect of interferon alfa (IFN-alpha) therapy on SENV replication, SENV DNA was quantitated by polymerase chain reaction in serum samples from 186 patients with chronic hepatitis C. Thirty-nine of 186 (21%) patients with chronic hepatitis C were positive for SENV DNA. There were no differences in the clinical, virologic and histologic features between patients with and without SENV infection. Eighteen of 102 patients with chronic hepatitis C who received IFN-alpha were positive for SENV DNA. The sustained response rate for hepatitis C virus (HCV) clearance after IFN-alpha treatment did not differ significantly between patients with SENV (28%) and without SENV infection (39%). SENV DNA levels decreased during therapy in 15 of 16 patients, and 11 of the 16 patients (69%) had a sustained loss of SENV DNA in response to IFN-alpha. In coinfected patients, SENV responses to IFN-alpha were significantly better in those who failed to clear HCV RNA than in those who lost HCV RNA (P =.013). In conclusion, SENV infection was frequently found in patients with chronic hepatitis C. SENV infection had no apparent influence on the severity of HCV-related liver disease or the HCV response to IFN-alpha. SENV was sensitive to IFN-alpha therapy and the majority of patients had a sustained virologic response. PMID- 11915045 TI - The allocation of livers for transplantation: a problem of Titanic consideration. PMID- 11915046 TI - Inflammation in response to hepatocellular apoptosis. PMID- 11915047 TI - Viral kinetics of hepatitis C: new insights and remaining limitations. PMID- 11915048 TI - Ursodeoxycholic acid cytoprotection: dancing with death receptors and survival pathways. PMID- 11915049 TI - High prevalence of HCV infection in patients with B-cell non-Hodgkin's lymphoma: comparison with birth cohort- and sex-matched blood donors in a Japanese population. PMID- 11915050 TI - Tissue typing for HFE mutations. PMID- 11915051 TI - Incidence of interferon alfa-induced depression in patients with chronic hepatitis C. PMID- 11915052 TI - Chronic hepatitis B guidelines: east versus west. PMID- 11915054 TI - There is nothin' like a Dame. PMID- 11915055 TI - Acute pain services revisited--good from far, far from good? PMID- 11915056 TI - Exploring the epidural trail. PMID- 11915057 TI - A survey of acute pain services in Germany and a discussion of international survey data. AB - BACKGROUND AND OBJECTIVES: Acute pain services (APS) have been introduced worldwide to improve postoperative pain management. However, data are lacking on organization and quality of these services in Germany. METHODS: A questionnaire on postoperative pain management was mailed to 773 departments of anesthesiology. Practice was compared between hospitals with and without APS and between services that did or did not meet basic quality criteria (QC): personnel assigned for APS, policies during nights and weekends, written protocols of pain management, and regular assessment and documentation of pain scores at least once a day. RESULTS: With 446 responding hospitals, 161 operated an APS. Large hospitals organized an APS more frequently than smaller hospitals (P <.001). Staff being assigned for the APS comprised anesthesiologists in 68.3% and nurses in 23.6% of the departments. In total, only half of the APS complied with basic QC. Regular assessment and documentation of pain scores were the QC most often missing (46.6%). Patient-controlled analgesia (PCA) was performed in 93.8%, 74.1%, and 69.8% of the departments providing APS with QC, APS without QC, and no APS. Epidural analgesia (EA) was available in nearly all departments (96.9%). A total of 75.6% of the departments performed EA on general wards (with QC: 93.8%, without QC: 82.7%, and no APS: 68.4%, P <.001). Frequency of PCA and EA use was highest in hospitals providing an APS with QC. CONCLUSIONS: As 50% of German APS do not comply with QC, we believe structured policies, as well as quality assurance measures, should be emphasized in the future. A comparison to international survey data showed that APS, in general, often do not consider QC. PMID- 11915058 TI - Epidural analgesia and postoperative lipid metabolism: stable isotope studies during a fasted/fed state. AB - BACKGROUND AND OBJECTIVES: Although previous studies have reported an inhibitory effect of epidural block and glucose feeding on plasma concentrations of glycerol and free fatty acids (FFA), it remains unclear how epidural analgesia modifies the postoperative production and uptake of lipid metabolites. This can be achieved by determining the rate of lipolysis during a feeding state with dextrose. METHODS: Twelve patients with or without postoperative epidural analgesia were studied 48 hours after surgery. They underwent a 6-hour stable isotope infusion study using [1,1,2,3,3,-(2)H(5)] glycerol; 3 hours of fasting, and 3 hours of dextrose infusion (4 mg/kg/min). The rate of glycerol appearance (R(a) glycerol) i.e., rate of lipolysis, and plasma concentrations of glycerol, FFA, glucose, lactate, insulin, glucagon, and cortisol were measured during the fasted and the fed states. RESULTS: The rates of lipolysis were similar in both groups during the fasted state and were not modified by dextrose infusion. In contrast, plasma concentrations of glycerol and FFA were decreased significantly during the fed state (P <.01). Glycerol clearance (ratio between R(a) glycerol and plasma glycerol concentration) increased significantly in both groups (P <.05) with feeding. Similarly, plasma concentrations of glucose and insulin increased significantly following feeding with dextrose in both groups. CONCLUSIONS: The elevated rates of lipolysis associated with surgery cannot be suppressed by either epidural analgesia or dextrose feeding implying that the sustained stress response continues in the postoperative period and is the most important factor responsible for the increased release of glycerol. PMID- 11915059 TI - The effect of single-injection femoral nerve block on rehabilitation and length of hospital stay after total knee replacement. AB - BACKGROUND AND OBJECTIVES: Continuous-infusion femoral nerve block (FNB) improves analgesia and rehabilitation after total knee replacement. In this study, we investigated the efficacy of single-injection FNB to achieve similar results. METHODS: A total of 30 patients were prospectively and randomly assigned to receive 40-mL injections of either 0.25% bupivacaine (group B) or saline (group S) after total knee replacement. Blinded observers evaluated the patients for postoperative pain, morphine consumption, ambulating distances, and maximal knee flexion; pain was scored on the visual analog scale (VAS). RESULTS: Compared with group S patients, group B patients had significantly lower VAS pain scores (P <.01 in the postoperative anesthesia care unit, P <.05 on the day after surgery); group B patients also showed significantly lower total morphine use (P <.05) and a lower incidence of morphine-related side effects. Significantly more group B than group S patients could ambulate on the day after surgery (93% v 46%, P <.05), and mean ambulatory distance was significantly better for group B than group S patients at discharge (166 +/- 37 v 117 +/- 24 feet, P <.01). Knee flexion was significantly better for group B than group S patients on the second day after surgery (70 degrees v 60 degrees, P <.01), but the between-group difference was no longer statistically significant at discharge. Mean length of acute hospitalization was significantly shorter for group B (3 days; range, 3 to 5 days) than group S patients (4 days; range, 3 to 6 days, P <.05). CONCLUSIONS: Single-injection FNB provided effective analgesia, facilitated early ambulation, and reduced the length of acute hospitalization in patients undergoing total knee replacement. PMID- 11915060 TI - The efficacy of continuous intrabursal infusion with morphine and bupivacaine for postoperative analgesia after subacromial arthroscopy. AB - BACKGROUND AND OBJECTIVES: The aim of this study was to determine whether an intrabursal morphine and bupivacaine mixed infusion provides useful analgesia for prolonged pain relief after subacromial arthroscopic operation. METHODS: A continuous intrabursal infusion catheter was inserted at the conclusion of the subacromial arthroscopic operation that was performed under general anesthesia. In a prospective, double-blind, randomized trial, 60 patients were divided into 2 groups: an anesthetic group received 5 mL of mixed 0.5% bupivacaine, 2 mg of morphine, 0.05 mL of 1/1,000 epinephrine as a bolus, and a solution of 40 mL of a 0.5% bupivacaine and 8 mg of morphine mixture that was used as a maintenance dose at a constant rate of 0.5 mL/h. This was done by means of a continuous infusion pump (0.5 mL hourly). A saline group (n = 30) received continuous saline infusion. Two patients were eliminated from the study because of catheter leakage or malfunction in the saline group. The intensity of the pain was evaluated preoperatively and postoperatively for 3 days by a graded visual analog scale (score from 0 to 10) for night pain, pain on motion, sleep disturbance, lying on painful shoulder, and the amounts of supplemental analgesics. RESULTS: Pain was decreased on the first and second postoperative day, and there was less sleep disturbance for 3 days postoperatively in the anesthetic group. There was no difference in pain caused by movement postoperatively. In the anesthetic group, lesser amounts of analgesics were used in the first 48 hours postoperatively. CONCLUSIONS: The continuous intrabursal infusion method resulted in a decreased perception of rest pain and reduced supplemental analgesics requirement for 2 days postoperatively. PMID- 11915061 TI - Distribution of solution in the epidural space: examination by cryomicrotome section. AB - BACKGROUND AND OBJECTIVES: The routes of distribution and barriers to flow of solutions in the epidural space are incompletely determined. This study examined macroscopic details of epidural injectate spread in postmortem humans by cryomicrotome imaging. METHODS: Soon after death, 3 nonembalmed adult human subjects were injected with ink through epidural catheters inserted by standard techniques. Following freezing, microtome sectioning was performed to reveal anatomic features down to 100 microm. To control for effects of death, an adult baboon was injected during general anesthesia and subsequently examined in the same fashion. RESULTS: Injected ink was readily evident and showed spread as rivulets through numerous small channels rather than as a unified advancing front. The fascia that extends laterally from the posterior longitudinal ligament is an important barrier restricting solution flow. Solution preferentially traveled along the nerve root sheath through the intervertebral foramen. CONCLUSIONS: Distribution of solution in the epidural space is nonuniform. Rather than a uniform advancing front, spread is directed among paths between structures according to pressures by which they are compressed. No structural barriers block flow through the intervertebral foramina or spinal canal other than the fascia of the posterior longitudinal ligament. PMID- 11915062 TI - Comparison of racemic bupivacaine, ropivacaine, and levo-bupivacaine for pediatric caudal anesthesia: effects on postoperative analgesia and motor block. AB - BACKGROUND AND OBJECTIVES: To compare ropivacaine, levo-bupivacaine, and racemic bupivacaine for caudal blockade in children. METHODS: Using a prospective observer blinded design, 60 sevoflurane anesthetized children (1 to 7 years) undergoing minor subumbilical surgery, were randomized to receive a caudal block (1 mL/kg) with either ropivacaine 0.2%, racemic bupivacaine 0.25%, or levo bupivacaine 0.25%. Postoperative analgesia (number of patients needing supplemental analgesia as defined by an objective pain score [OPS] score of > or = 5; time to first analgesic demand) during the first 24 postoperative hours was chosen as the primary end-point. Early postoperative motor block (3-point scale) was assessed as a secondary end-point. RESULTS: All blocks were judged to be clinically successful based on the presence of adequate intraoperative and early postoperative analgesia. An OPS score > or = 5 was found in 5/20 patients in each study group. No difference regarding the time to first analgesic demand was found between the study groups. The use of ropivacaine (P =.02), but not levo bupivacaine (P =.18), was found to be associated with less motor block during the first postoperative hour compared with racemic bupivacaine. CONCLUSION: All 3 investigated local anesthetics were found to be clinically comparable despite the slight reduction of early postoperative motor block associated with the use of ropivacaine. PMID- 11915063 TI - Buprenorphine added to the local anesthetic for axillary brachial plexus block prolongs postoperative analgesia. AB - BACKGROUND AND OBJECTIVES: Buprenorphine added to local anesthetic solutions for supraclavicular block was found to triple postoperative analgesia duration in a previous study when compared with local anesthetic block alone. That study, however, did not control for potentially confounding factors, such as the possibility that buprenorphine was affecting analgesia through intramuscular absorption or via a spinal mechanism. To specifically delineate the role of buprenorphine in peripherally mediated opioid analgesia, the present study controlled for these 2 factors. METHODS: Sixty American Society of Anesthesiologists (ASA) P.S. I and II, consenting adults for upper extremity surgery, were prospectively assigned randomly in double-blind fashion to 1 of 3 groups. Group I received local anesthetic (1% mepivacaine, 0.2% tetracaine, epinephrine 1:200,000), 40 mL, plus buprenorphine, 0.3 mg, for axillary block, and intramuscular (IM) saline. Group II received local anesthetic-only axillary block, and IM buprenorphine 0.3 mg. Group III received local anesthetic-only axillary block and IM saline. Postoperative pain onset and intensity were compared, as was analgesic medication use. RESULTS: The mean duration of postoperative analgesia was 22.3 hours in Group I; 12.5 hours in group II, and 6.6 hours in group III. Differences between groups I and II were statistically significant (P =.0012). Differences both between groups I and III and II and III were also statistically significant (P <.001). CONCLUSIONS: Buprenorphine-local anesthetic axillary perivascular brachial plexus block provided postoperative analgesia lasting 3 times longer than local anesthetic block alone and twice as long as buprenorphine given by IM injection plus local anesthetic-only block. This supports the concept of peripherally mediated opioid analgesia by buprenorphine. PMID- 11915064 TI - Continuous subgluteus sciatic nerve block after orthopedic foot and ankle surgery: comparison of two infusion techniques. AB - BACKGROUND AND OBJECTIVE: To compare continuous infusion or a patient-controlled technique for postoperative analgesia after foot surgery, using a new subgluteus approach for continuous sciatic nerve block. METHODS: Fifty healthy patients, undergoing orthopedic foot surgery, received a continuous sciatic nerve block using a new subgluteus approach. All blocks were placed with the aid of a nerve stimulator using a 10-cm, 18-gauge insulated Tuohy needle. After either plantar flexion or dorsiflexion of the operated foot was elicited at < or = 0.5 mA, 20 mL of 0.75% ropivacaine was injected incrementally using repeated aspiration tests, then followed by the introduction of a 20-gauge epidural catheter. Postoperatively, 0.2% ropivacaine was infused with either a 10 mL/h continuous infusion (group Continuous, n = 25) or with a 5 mL/h basal rate with 5 mL bolus every 60 minutes (group patient-controlled analgesia [PCA], n = 25). Intraoperative analgesic supplementation, as well as postoperative pain relief, morphine consumption, incidence of complication, and patient satisfaction were recorded by an observer unaware of group assignment. RESULTS: The sciatic catheter was successfully placed in all patients. Intravenous fentanyl supplementation (dose range, 50 to 150 microg) was required in 4 patients in each group, but no patient required general anesthesia. Catheter dislocation was reported in 2 patients (4%). The quality of pain relief was good in both groups, and none experienced complications. Nine patients of the Continuous group (37%) and 7 patients of the PCA group (29%) required rescue morphine analgesia because of pain in the femoral dermatomes (P =.76). Ropivacaine consumption was 240 mL in the Continuous group (range, 200 to 240 mL) and 140 mL in the PCA group (range, 120 to 290 mL) (P =.0005). Patient acceptance was good in both groups. CONCLUSIONS: The continuous subgluteus sciatic nerve block represents an easy and reliable option for postoperative analgesia after foot surgery; using a patient controlled rather than a continuous infusion technique reduces the consumption of local anesthetic solution without affecting the quality of pain relief. PMID- 11915065 TI - Use of a charged lidocaine derivative, tonicaine, for prolonged infiltration anesthesia. AB - BACKGROUND AND OBJECTIVES: We tested the hypothesis that the duration of cutaneous anesthesia elicited by the permanently charged compound N-phenylethyl lidocaine (tonicaine) would be longer than that elicited by its parent structure, lidocaine, and that it would be less affected by epinephrine (epi), after subcutaneous injection in rats, as a model for infiltration anesthesia. METHODS: Subcutaneous injections were performed on the shaved dorsal skin of rats with either tonicaine or lidocaine (0.1% or 0.5%, n = 8 in each group) with and without epi (1:200,000). Inhibition of the cutaneous trunci muscle reflex was quantitatively evaluated by a blinded observer by the number of times pinpricks failed to elicit the nocifensive motor response out of a total of 6 pinpricks applied to the injected area. RESULTS: Duration of complete nociceptive blockade in the 0.5% tonicaine and lidocaine groups was 619 +/- 47 and 58 +/- 2 minutes, respectively; duration of full recovery in these groups was 1,106 +/- 19 and 86 +/- 3 minutes, respectively. Epi increased the duration of complete block in the 0.5% tonicaine and lidocaine groups to 750 +/- 13 and 97 +/- 11 minutes, respectively, and the duration of full recovery to 1,185 +/- 13 and 172 +/- 6 minutes, respectively. Skin toxicity was seen only in the 0.5% tonicaine with epi group (3 of 8 rats). CONCLUSIONS: Tonicaine is a substantially longer lasting local anesthetic with a delayed onset of action compared with lidocaine and may be useful in situations where long duration of infiltration block is desirable. PMID- 11915066 TI - Airway regional anesthesia for awake fiberoptic intubation. PMID- 11915067 TI - The anesthesiologist's guide to personal digital assistants. AB - BACKGROUND AND OBJECTIVES: Personal digital assistants (PDAs) are being used more frequently by anesthesiologists. PDAs are pocket-sized electronic devices that, at their most basic level, store information. Basic features include calendar, phone directory, memo pad, and checklist. More advanced features are increasingly common and blur the distinction between PDAs and personal computers (PCs). This article reviews the main options for PDAs, purchasing tips, and software applications, especially as they apply to anesthesiologists. PMID- 11915068 TI - Analgesia administration attenuates surgery-induced tumor promotion. PMID- 11915069 TI - Ultrasound imaging of the thoracic epidural space. AB - BACKGROUND AND OBJECTIVES: In thoracic epidural anesthesia, the "loss of resistance" technique is the standard technique for the identification of the epidural space (EDS), the feedback to the operator is often solely tactile. Our aim was to establish ultrasonography for the prepuncture demonstration of the anatomic structures surrounding the thoracic EDS and to evaluate its precision and imaging quality. METHODS: We examined 20 volunteers. In each participant, the extradural space and the neighboring anatomic landmarks in the intervertebral space Th 5-6 were identified using 2 imaging techniques: magnetic resonance imaging (MRI) and ultrasonography. We compared corresponding images regarding distance measurements and the visibility of anatomic landmarks. RESULTS: The capacity of ultrasound imaging (US) to depict the thoracic EDS was limited. Due to the better overview, MR images were easier to interpret. However, US proved to be of better value than MRI in the depiction of the dura mater. All important landmarks for the puncture of the thoracic EDS could be identified with both techniques. The overall correlation was satisfactory. US depicted the different structures of the thoracic EDS with an acceptable precision (confidence interval, 4.6 to 8.7 mm). CONCLUSIONS: US showed good correlation with MRI, which is a standard imaging technique for the depiction of the spine. We anticipate that prepuncture ultrasonography may facilitate thoracic epidural anesthesia by needle placement. PMID- 11915070 TI - Estimating with confidence the risk of rare adverse events, including those with observed rates of zero. AB - Omission of a confidence interval (CI) associated with the risk of a serious complication can lead to inaccurate interpretation of risk data. The calculation of a CI for a risk or a single proportion typically uses the familiar Gaussian (normal) approximation. However, when the risk is small, "exact" methods or other special techniques should be used to avoid overshooting (risks that include values outside of [0,1]) and zero width interval degeneration. Computer programs and simple equations are available to construct CIs reasonably accurately. In the special case in which the complication has not occurred, the risk estimated with 95% confidence is no worse than 3/n, where n is the number of trials. PMID- 11915071 TI - Chronic pain with beneficial response to electroconvulsive therapy and regional cerebral blood flow changes assessed by single photon emission computed tomography. AB - BACKGROUND: Recent neuroimaging studies suggested that chronic neuropathic pain may be largely sustained by a complex neuronal network involving the thalamus. Although recent studies have demonstrated the efficacy of electroconvulsive therapy (ECT) in the treatment of a variety of types of chronic neuropathic pain, the effects of ECT on regional cerebral blood flow (rCBF) have not been studied. OBJECTIVES AND METHODS: We present a 50-year-old female postsurgical chronic pain patient whose pain had failed to respond to standard pain treatment, but was resolved by ECT. To investigate the potential role of rCBF in ECT's analgesic effect, we measured significant changes in the rCBF in the thalamus before and after a course of bilateral ECT using technetium-99m ethyl cysteinate dimer (99mTc-ECD) single photon emission computed tomography (SPECT). RESULTS: 99mTc ECD SPECT showed a significant bilateral decrease in the thalamus on the side of the pain, and this decreased rCBF in the thalamus increased after ECT. CONCLUSIONS: The results from the SPECT suggest that ECT increases abnormally decreased thalamus activity in chronic neuropathic pain. PMID- 11915072 TI - Foraminal injection of a painful sacral nerve root using an epidural catheter: case report. AB - BACKGROUND AND OBJECTIVES: Nerve root pain has been treated with steroid injections since the 1970s. We will describe a novel method for nerve root injection using a directed epidural catheter. CASE REPORT: A case is described where catheter placement accurately reproduced symptoms which were then relieved with local anesthetic and steroid. CONCLUSION: This is a novel approach to treatment of painful nerve roots with a catheter. PMID- 11915073 TI - Patient-controlled epidural analgesia for labor and delivery in a parturient with chronic inflammatory demyelinating polyneuropathy. AB - OBJECTIVE: The anesthetic management of labor and delivery in patients with any form of chronic inflammatory demyelinating polyneuropathy (CIDP) is not well defined. Using patient-controlled epidural analgesia (PCEA), or epidural analgesia, in such a rare clinical situation has not been previously reported. CASE REPORT: A 32-year-old, gravida 3, para 2, woman with a 2(1/2) year history of CIDP was admitted for labor and delivery at 38 weeks of pregnancy. At the time she presented for labor analgesia, she complained of bilateral hand and foot weakness and tingling. PCEA with 0.1% ropivacaine and fentanyl 2 microg/mL was used for labor analgesia. The patient was pain free during labor and delivery and had an uneventful postpartum course. CONCLUSION: PCEA had no apparent detrimental affect on the patient's disease and may be a reasonable option for patients with CIDP presenting for labor and delivery. PMID- 11915074 TI - Vertebral osteomyelitis and psoas abscess occurring after obstetric epidural anesthesia. AB - BACKGROUND AND OBJECTIVES: Back pain and infectious complications occasionally occur after epidural anesthesia in obstetrics, and accurate diagnosis can be difficult. We report a patient who developed low back pain soon after obstetric epidural anesthesia and was diagnosed 6 months later with lumbar vertebral osteomyelitis, discitis, and a psoas abscess. CASE REPORT: A 34-year-old woman developed persistent low back pain after receiving epidural anesthesia for labor analgesia and cesarean delivery. After 6 months, a diagnosis of lumbar vertebral osteomyelitis, discitis, and psoas abscess was made, and surgery was performed. Because of the temporal and anatomical relationships between epidural catheterization and the development of symptoms, the preceding epidural anesthesia was initially suspected as a potential cause. However, because the posterior spinal elements were unaffected and the infectious agent was subsequently identified as tuberculous, the cause was eventually determined as unlikely to be related to the epidural procedure. CONCLUSION: Investigation of severe back pain after epidural anesthesia should include consideration of infectious causes, such as vertebral osteomyelitis and discitis, which may not be causally related to the epidural catheterization itself. PMID- 11915075 TI - Preemptive multimodal analgesia for anterior cruciate ligament surgery. PMID- 11915079 TI - How close is close enough--how close is safe enough. PMID- 11915077 TI - Combined lumbar and sacral plexus block for the management of long-standing hip pain. PMID- 11915081 TI - If you can chart, you can write. PMID- 11915082 TI - Surfing the eWave. PMID- 11915083 TI - Serving God through CNE. PMID- 11915084 TI - Public speaking. A tool God can use. PMID- 11915085 TI - A mentoring relationship. Two-nurses grow in faith & knowledge. PMID- 11915086 TI - A Christian model of health promotion. PMID- 11915087 TI - Josh's time. PMID- 11915088 TI - Wide shoulders of care. PMID- 11915090 TI - She saw Jesus. Discovering the living word. PMID- 11915089 TI - The touch of a nurse friend. PMID- 11915091 TI - Telehealth: an opportunity for gerontological nursing practice. PMID- 11915092 TI - How accurate are chronic wound assessments using interactive video technology? AB - This project examined the accuracy of chronic wound assessments made using an interactive, video telecommunications system (Teledoc 5000, NEC America, Inc., Irving, TX) by comparing a nurse expert's in-person wound assessments with wound assessments made from taped Teledoc sessions. Wound assessments determined the absence or presence of nine wound characteristics instrumental in guiding treatment (e.g., tunneling, undermining, granulation tissue, necrotic tissue, epithelial tissue, purulent exudate, erythema, edema, induration). A sample of 13 paired wound observations was analyzed. The accuracy of the Teledoc technology was examined by calculating the amount of agreement between the in-person assessments and the taped Teledoc assessments for each of the nine characteristics. Agreement for eight of the nine wound characteristic exceeded 75%, suggesting this telehealth medium does not alter wound assessment data, which are essential in guiding treatment decisions. In addition to connecting the remotely based nurse with nursing expertise to improve patient care, telehealth technology seemed to increase the remotely-based nurses' knowledge of wound assessment and treatment as well. PMID- 11915093 TI - Assessing elderly patients with congestive heart failure via in-home interactive telecommunication. AB - The efficacy of telemedicine technology was tested for equivalence of nursing assessment with chronic congestive heart failure (CHF) home care patients (N = 28). The equivalence of nurses' physical assessment findings was estimated using an innovative two-way, telemedicine audiovisual system. Nurses were randomly assigned to a method of client assessment: on-site (real time) or telemedicine (monitor time). Each assessment was performed within 10 minutes of each other. Assessment variables compared were auscultation of lung sounds, heart sounds, rate and rhythm, blood pressure, weight, edema, respiratory effort, and client's face, lip, and nail color. Eighteen physiological parameters were analyzed, using either the Wilcoxon signed ranks test or the McNemar test. Results indicate few significant differences between the assessments of the real time and monitor time nurses. The monitor nurse was more likely to claim abnormality than the real nurse was when assessing the color of nails (p = .048). The real nurse picked up ankle edema (p = .024), pedal edema (p = .099), and inspiratory wheeze (p = .01) more frequently than did the monitor nurse. Kappa coefficients to determine the extent of agreement between nurse's assessments were significant. Nurses' comments were favorable, but they recommended altering the interview to elicit symptoms not easily observed by the monitor nurse such as diaphoresis. Exit interviews of the elderly patients revealed a favorable reaction to using the telemedicine monitor, citing a quick connection to a nurse and response to their concerns and questions. Both nurses and patients reported the need to have real nurse home visits along with telemedicine visits. PMID- 11915094 TI - A nursing application of telecommunications. Measurement of satisfaction for patients and providers. AB - Consumer and provider satisfaction is key to the continued use and expansion of telehealth technology. This pilot study compared satisfaction of providers and patients with wound consultations done in person with those done via real-time interactive video technology. Eleven telehealth consultations with a nurse expert were immediately followed by an in-person consultation with a second nurse expert. Satisfaction questionnaires were administered to patients, referring nurses, and the consultant nurse expert following both the in-person consultation and the telehealth consultation. The referring nurses (100%) were satisfied with both the telehealth and in-person consultations, noting the ability to provide better care for their patients. The patients (55%) were "very satisfied" with the telehealth consultations versus 40% satisfied with the in-person consultations. Difficulty in hearing for the patients was equal in both groups, which resulted in changes in the consultation process. The patients' difficulty in seeing the telehealth consultant was addressed through larger screens and strategic positioning to provide easier viewing for the patient and providers. The telehealth nurse consultant was satisfied overall but had some difficulty communicating. This pilot study helped provide useful information for both the telehealth and in-person consultations. PMID- 11915095 TI - Evaluating the cost of one telehealth application connecting an acute and long term care setting. AB - This article describes a study of the costs of a pilot telemedicine chronic wound consultation clinic. Cost minimization analysis is the technique used to examine the costs of the clinic. The components of cost analysis include the fixed costs of personnel and equipment and the indirect costs of circuit and line charges. Cost avoidance is also examined. Cost avoidance evaluates what costs were avoided by the use of the telemedicine clinic. Additionally, the cost perspectives of the consulting agency, the referring agency, and the patient are examined. The average cost of a chronic wound consultation was $136.16 (acute care perspective). Costs of a traditional face-to-face consultation, if the residents were transported to the acute care facility would be $246.28. Fifteen telehealth consultations per month were used to determine per consultation costs for line charges and depreciation/maintenance costs. In this pilot study, a cost savings was realized and patients benefited. Increased volume will help to offset the cost of the equipment depreciation and maintenance and make telehealth chronic wound consultations more cost effective. PMID- 11915096 TI - Determinants of preventive services utilization in rural older women. AB - Research describing preventive services utilization and determinants of preventive services utilization for rural older women is scant. The purpose of this study was to: (a) describe the prevalence of preventive services utilization in compliance with the U.S. Preventive Services Task Force (USPSTF) recommendations for screening, counseling, and immunizations among rural older women in four Nebraska counties and (b) determine the extent to which personal influences (e.g., demographics, definition of health, perceived health status) and contextual influences (e.g., access to care, sources of health information, provider recommendations) explain preventive services utilization among those women. A sample of 102 community-dwelling rural women age 65 and older participated in the study. The prevalence of receipt of USPSTF-recommended preventive services utilization among these rural women was alarmingly low. Multiple regression analyses revealed that provider recommendations, access to care, sources of health information, and perceived health status were significantly associated with preventive services utilization. These findings provide information that is relevant in designing interventions to increase preventive services utilization among rural older women. PMID- 11915097 TI - Helping older adults at home: public policy and home care. PMID- 11915098 TI - A new room for Aunt Ruth. PMID- 11915099 TI - Staffing ratios and research. PMID- 11915100 TI - Formal caregiving in home care. PMID- 11915101 TI - Telehealth. PMID- 11915102 TI - Circle of Christian caring. A model for parish nursing practice. PMID- 11915103 TI - What's a parish nurse to do? Congregational expectations. PMID- 11915104 TI - Church health fairs. Partying with a purpose. PMID- 11915105 TI - Breaking the cycle of school violence. How can parish nurses help? PMID- 11915106 TI - Spiritual assessment. Comparing the tools. PMID- 11915107 TI - The power of presence. PMID- 11915108 TI - Parish nursing: firing the imagination. PMID- 11915109 TI - Paulette Holly. Haitian nurse, deaconess, exemplar. PMID- 11915110 TI - Conversations with God. Keeping a prayer journal. PMID- 11915111 TI - To stop or not. Good Samaritan 2001. PMID- 11915112 TI - Speak carefully. PMID- 11915113 TI - Parish nursing. Jesus people style. PMID- 11915114 TI - Inner City Health Center. Dispensing help & hope. PMID- 11915115 TI - Aging, ethnicity, social support. A review--part 1. AB - The increasing diversity of the population of older adults is reflective of the worldwide demographic trends. These trends present nurses with special challenges in assessing and meeting the needs of ethnic elderly individuals. Nurses must be willing and capable of assessing the social support networks and beliefs of older adults from any ethnic group to plan and implement culturally sensitive care. The success of this care will be closely related to the extent to which nurses recognize and include existing support networks and individual preferences. PMID- 11915116 TI - Relocation among ethnic elders. A review--part 2. AB - Relocation to more supportive housing is a potentially stressful life event for older adults. Under the best of circumstances, the older adult will be relatively healthy and able and willing to participate in relocation decisions. However, this is not always possible. Nurses have a critical role to play in helping families identify the most appropriate housing alternative, helping plan the relocation, and helping older adults adjust to their new homes. Relocation is particularly stressful for ethnic elders who may not understand the housing options available, or may fear prejudicial treatment and exclusion in the various settings. Nurses need to take time for careful assessment of the older adults' preferences, and to provide information needed to help them make relocation decisions within the context of their existing support networks. PMID- 11915117 TI - Exploration of spirituality & health among diverse rural elderly individuals. PMID- 11915118 TI - The 20th century. Looking back at the ambiance of aging from the perspective of age-specific journals and periodicals. AB - The word "aging" is fraught with nuance, both positive and negative. The purpose of this article is to describe the results of a preliminary exploration of the ambiance of the concept of aging in the United States as seen from the perspective of periodical literature from the past century. A convenience sample of professional journals and lay periodicals concerned with aging and the issues of aging was selected as the focus for this investigation. Specifically, volume one, number one of the selected journals was examined with regard to the date of the first publication, the table of contents, and the first issue editorial or articles in the first volume describing the motivation for the introduction of the journal. Findings indicated that as the century progressed and the study of aging took form, the focus shifted from how to manage the problems of aging to: (a) how to promote healthier young adult lifestyles to increase the chances of a healthy old age, (b) how to continue to maintain older adults' health and therefore, increase the life expectancy for those people older than age 65, and (c) how older adults contribute to the well-being of society. The longevity of the human species, as well as the increasing size of the aging population is a new phenomenon, the ramifications of which are difficult to predict. However, as the emotional tone of the journals in the latter quarter of the century gradually became more positive, it was apparent that consideration was being given to the idea that this phenomenon could be a positive force in the collective growth of humanity. PMID- 11915119 TI - Teaching elderly individuals on computer use. PMID- 11915120 TI - Nurses who care for survivors of the Holocaust. Is special education necessary? PMID- 11915121 TI - Safety vests: helping patients who wander to find their way home. PMID- 11915122 TI - Detection of depression in the cognitively intact older adult protocol. PMID- 11915123 TI - Births, marriages, divorces, and deaths: provisional data for July 2001. PMID- 11915124 TI - Intermittent malaria treatment as a component of the EPI (Expanded Programme on Immunization) schedule in Africa. PMID- 11915126 TI - Marion Milner, mysticism and psychoanalysis. AB - It is unusual to combine mysticism and psychoanalysis. Marion Milner, however, achieved precisely this. Through her self-analysis and analytic work with children and adults--and using as an illustration her own and others' imaginative ideas, paintings, doodles, drawings and pictures--she drew attention to the potential for health and creativity of undoing the obstacles to mystical experience of oneness with what is beyond or other than the self, which she sometimes called God, the unconscious or the id. This article seeks to explain and highlight this aspect of her contribution to, and continuing importance for, psychoanalytic theory and practice--particularly that associated with Winnicott- through detailing her early life and diary-keeping experiments, some of her psychoanalytic case histories during and after the Second World War, her work as an artist, ending with her travels and her involvement during the 1980s and 1990s with the Squiggle Foundation and British Association of Art Therapists. PMID- 11915127 TI - Non-discursive expressive elements and their role in the construction of meaning in the analytical situation. A clinical discussion. AB - The author discusses the role of non-discursive expressive elements in the construction of the analytical situation, using three examples to illustrate the problems with which he is concerned. His claim is that the issue in question necessarily involves the subject of affects, and he proceeds to discuss the difficulties associated with this subject. In addition, he considers the contributions of Green and Imbasciati, and Kleinian developments of this theme, including also the contributions of Bion--in particular the latter's theories concerning thought, in which emotion comes to assume an essential place in the origin of thinking. The author resumes the discussion by taking up his clinical examples, using them to put forward the view that non-discursive expressive elements may well play a decisive role in the construction of meaning in the analytical situation. He suggests also that the meaning of an emotional experience may be thought of as a construction contributed to by a number of symbolic forms which both interfere with and interact with the symbolic system of language. Following examination of his third example, the author reflects on 'musicality', a notion sometimes referred to informally in clinical data in connection with the 'emotional climate' of the session. He proposes that the complex problem of meaning in music be extended to cover the construction of meaning in the psychoanalytic setting, and in so doing returns to ideas put forward by Suzanne Langer. His underlying view here is that essential elements of the musical phenomenon and essential elements of particular forms of emotional life give rise to the same emotional matrices--perhaps to what Meltzer calls 'musical deep grammar'. Finally, the author considers various symbolic forms that contribute to the particular configuration of analytical situations, suggesting that the mental condition of 'free-floating attention' requires the broad availability to the analyst's mind of a multiplicity of symbolic forms, his conversion of these into new expressions of meaning, and the possibility of their verbal communication by him to the patient. PMID- 11915128 TI - 'Never twice without thrice'. An outline for the understanding of traumatic neurosis. AB - Following the review of Freud's work on trauma (1916-17, 1920, 1926, 1933, 1939), this article proposes to view the traumatic scene as a screen which acts as an active process in the case of many victims who, like Mrs D, break down a few days after being assaulted. After a first attempt at denial, the ego is submerged with repressed content. Repetition is seen as a means of abreacting the trauma by seeking to bind the enormous quantity of excitations that cannot find discharge. Repetition, because of its compulsive quality, fails. The victim remains in a state of helplessness against which she resorts to denial, splitting and projection (Freud, 1920, 1926). This strategy is seen, however, as maintaining the victim in a state of helplessness. Thus, I suggest that, overwhelmed, the ego finally resorts to a multi-levelled system of defence. On a first level, fixation to the trauma, like in screen-memories, gives way to a first displacement. On a second level, the adoption of an identity built upon being a victim contributes to the strength of this displacement. On a third level, identification with the aggressor consolidates this defensive strategy by giving meaning to the hostility awakened by the sexual assault. PMID- 11915129 TI - Wittgenstein's personality and his relations with Freud's thought. AB - In this contribution the author examines the connections between Wittgenstein's personality and his attitude to Freud's psychoanalytic theories in the light of biographies of the philosopher, published exchanges of letters between him and his sisters, his 'secret' diaries from the time of the First World War, his diaries from the nineteen-thirties and the writings in which he discusses Freud and psychoanalysis. The paper quotes liberally from all these sources. Following an account of Wittgenstein's cultural and family background in Vienna and his subsequent peripatetic life, hypotheses are presented concerning his personality, sexuality and 'internal' theology, together with some ideas about his relationship with his family (in particular, his parents and sisters) and his critique of Freud's theories, with particular reference to dreams and their interpretation. Wittgenstein emerges as a highly original philosopher who is, however, emotionally disturbed and restless. His personality is found to have narcissistic aspects that moulded his behaviour and thought, and the author contends that his mental suffering caused him to apply psychological and psychoanalytic categories to his philosophy. PMID- 11915131 TI - Presidential address. 42nd Congress of the International Psychoanalytical Association. PMID- 11915130 TI - A terminating case. PMID- 11915132 TI - Creating the psychoanalytical process incorporating three panel reports: opening the process, being in the process and closing the process. PMID- 11915133 TI - Psychoanalysis and allied therapies: what does a psychoanalyst do when he/she thinks or says he/she is doing psychotherapy? PMID- 11915134 TI - Neurosciences and psychoanalysis. PMID- 11915135 TI - The discourse and the psychoanalytic method. PMID- 11915136 TI - Changing psychoanalytic psychotherapy into psychoanalysis. PMID- 11915137 TI - Changing nature of family structure: developmental theories under challenge. PMID- 11915138 TI - In-patient treatment of substance abuse. PMID- 11915139 TI - New perspectives from an overview of the congress. PMID- 11915140 TI - Colour and music: voices of the unconscious. PMID- 11915141 TI - Reply to Norman and Thomson-Salo. PMID- 11915142 TI - A final response to Dr Flink. PMID- 11915143 TI - Saving the International Review? PMID- 11915144 TI - The rebirth of the idols: the Freudian unconscious and the Nietzschean unconscious. PMID- 11915145 TI - Psychoanalysis without Descartes. PMID- 11915146 TI - William Gillespie (1905-2001). PMID- 11915147 TI - Remembering a depressive primary object: memory in the dialogue between psychoanalysis and cognitive science. AB - Memory has always been a central issue in psychoanalytic theory and practice. Recent developments in the cognitive and neural sciences suggest that traditional notions of memory based on stored structures which are also often underlying psychoanalytic thinking cannot account for a number of fundamental phenomena and thus need to be revised. We suggest that memory be conceived as a) a theoretical construct explaining current behaviour by reference to events that have happened in the past. b) Memory is not to be conceived as stored structures but as a function of the whole organism, as a complex, dynamic, recategorizing and interactive process, which is always 'embodied'. c) Memory always has a subjective and an objective side. The subjective side is given by the individual's history, the objective side by the neural patterns generated by the sensory motor interactions with the environment. This implies that both 'narrative' (subjective) and 'historical' (objective) truth have to be taken into account achieving stable psychic change as is illustrated by extensive clinical materials taken from a psychoanalysis with a psychogenic sterile borderline patient. PMID- 11915148 TI - The relationship between mothering in infancy, childhood experience and adult mental health: results of the Brody prospective longitudinal study from birth to age 30. AB - This longitudinal study has followed seventy-six individuals from birth to age 30 using films of the mother-child interaction, psychoanalytically informed interviews of parents and children, and psychodiagnostic testing to assess how the quality of mothering a child receives in the first year of life contributes to his/her subsequent emotional well-being. The thirty-year follow-up of the now adult participants found that those who had received more effective care in infancy in terms of maternal empathy, consistency, control, thoughtfulness, affection and management of aggression had higher-level psychological defence mechanisms as adults than children receiving less effective nurturing--suggesting a process in which the children internalised their mothers' own defence mechanisms. Other measures at 30 years (Global Functioning, Erikson psychosocial attainment, mental representation of security of attachment to parents and presence or absence of a psychiatric diagnosis) did not achieve statistical significance. On the other hand, trauma after infancy and before age 18 provided the strongest correlation with adult outcome: consistent with the theory of the effect of cumulative trauma on psychic functioning, children experiencing two or more adverse circumstances had significantly lower levels of global functioning as adults than those spared multiple traumas. With case examples, the findings illustrate how the effect of pre-verbal experience attenuates over time, and how later influences overlie early life in the course of psychological structuralisation. PMID- 11915149 TI - 'The crisis in psychoanalysis': what crisis are we talking about? AB - In this paper the author argues that the so-called crisis in psychoanalysis, often blamed on various external factors, is in fact an internal crisis brought about by intrinsic incongruities between the explicit intention of its educational model, which aspires to educate and train in a professional and scientific discipline, and its organisational structure, locally and internationally inextricable. Its isolated basic units of ecumenical control--its traditional 'societies/institutes of psychoanalysis'--implicitly and explicitly co-impose the monastic transmission of a preponderantly doctrinaire education and clinical practice. The dysfunctional elongation of our historical organisational syncretism continues to force us, one century later, to amalgamate in and superimpose on an endogamous supraordaining system (i.e. our International Psychoanalytic Association) prerogatives and functions so contradictory that they are ordinarily considered irreconcilable, such as: education and scientific research (tasks of universities); societal and political (tasks of an ordinary society of professionals and technicians); and 'as if' certification and accreditation (tasks of external, local, interinstitutional collegiate bodies and independent, multirepresentative, national coalitions and consortia, as well as of local governmental legislation). The pervasive collective regressive phenomena derived from this homogamous and secluding organisational and educational syncretism has had a retardatory impact along all the hierarchy of our institutional activities. The future of psychoanalysis as a science and a clinical discipline must be nothing other than one of evolution and transformation. The survival of its legendary psychoanalytic institutes and societies, as well as its local and international organisation, with its inherited but now untenable syncretism, is that which is being questioned (that is, psychoanalysis as a 'movement' and a 'cause'). PMID- 11915150 TI - Psychoanalysis and infertility. Myths and realities. AB - Psychoanalysis, long interested in infertility, and a valuable treatment for men and women suffering with this affliction, has also helped to create and support a myth of psychogenic infertility. Multiple causes of infertility exist across the physiological-psychological spectrum. There is no simple psychodynamic causality. Advances in assisted reproductive technologies provide treatments that create emotional stress and outpace psychological preparedness of patients and analysts. This paper is based on the experience of a unique study group in Boston. An analytic case illustrates some of the ways analysis can be a treatment of choice for people using assisted reproduction. In fact, analysis offers a unique opportunity to elaborate fully the complex realities and dilemmas faced by people and their therapists throughout the infertility experience. More generally, this study of the concept of psychogenic infertility explores a valuable role for psychoanalysis in the treatment of medical conditions. PMID- 11915151 TI - Acute confusion: advancing clinical nursing science through research collaboration. PMID- 11915152 TI - Delirium in elderly patients: an overview of the state of the science. AB - Delirium is a common and potentially preventable and reversible cause of functional disability, morbidity, mortality, and increased health care use among elderly individuals. Much has been learned about delirium in the past decade. Highlighted in this article are recent advances in the diagnosis of delirium, delirium in long-term care, use of health care resources, outcomes of delirium, etiologies, and interventions to prevent and treat delirium. Suggestions for future research also are proposed. PMID- 11915153 TI - Acute confusion/delirium protocol. AB - This abbreviated version of the Acute Confusion/Delirium Research-Based protocol provides clinical guidelines for the assessment and management of acute confusion/delirium in the elderly individual. A screening and ongoing surveillance program that is based on identified risk factors is recommended to prevent or minimize episodes of acute confusion in this age group. This protocol is part of a series of protocols developed to help clinicians use the best evidence available in the care of older adults. PMID- 11915155 TI - Studying acute confusion in long-term care: clinical investigation or secondary data analysis using the minimum data set? AB - Clinical investigations of acute confusion have largely been initiated in the acute care setting, where no uniform patient assessment exists. No reliable estimates of the prevalence of acute confusion in long-term care (LTC) residents have been reported. Delirium indicators are present in the nursing home Minimum Data Set (MDS), suggesting that MDS assessments could be used to facilitate studies of acute confusion in LTC. Methods to study acute confusion in LTC are discussed, with an emphasis on the advantages and disadvantages of using secondary analysis of MDS assessments as one research strategy. PMID- 11915154 TI - Training of acute confusion resource nurses. Knowledge, perceived confidence, and role. AB - Most nurses function as generalists; however, some function in "expert" roles based on informal training as Resource Nurses. Training usually focuses on assessment and management of a specific problem, with the goal of creating a readily available "expert" for every unit. The primary activity of the Resource Nurse is to provide expert care, education, and consultation for patients, families, and staff. The Iowa-Veterans Affairs Nursing Research Consortium (IVANRC) addressed the need to manage acutely confused/delirious clients by training staff nurse volunteers (N = 129) from all units of the four Iowa Veterans Affairs facilities to act as unit-based acute confusion Resource Nurses (ACRNs). A day-long workshop included didactic content addressing etiology and presentation of acute confusion (AC), use of the IVANRC protocol to assess for AC, and basic information on treatment and management of AC. The nurses also participated in an efficacy-based experiential learning program on AC assessment that involved demonstrating assessment of AC and role enactment practice exercises in which ACRNs practiced the assessment. A test of knowledge of AC and perceived level of confidence in assessing acutely confused patients was administered before and after completion of the program. Paired t tests comparing pre- and posttest scores showed that knowledge and confidence significantly increased for the nurses as a result of their participation in the educational program. Eighteen months later, a second program was conducted to update current ACRNs and train additional RNs to enact this role. Pre- and posttest scores were obtained, with paired t tests showing a significant increase in knowledge for the participants. Twenty-four (49%) of the second program attendees had attended the first program. These participants had significantly higher scores on the second program pretest than those participants who had not attended the previous program, indicating a retention of knowledge from the first program. PMID- 11915156 TI - Acute confusion in terminally ill hospitalized patients. AB - Knowledge about acute confusion (AC) has grown rapidly during the past decade, but very few studies have focused specifically on AC episodes associated with the end of life. Although experienced oncology clinicians accept that AC is common near the end of life, little is known about the frequency, nature, course, and timing of AC during this critical stage of life in patients with terminal cancer. Data suggest patients with advanced cancer have reversible causes of delirium, where appropriate treatment can result in improved outcomes. The data for this article are drawn from a larger study investigating the incidence, prevalence, behaviors, and outcomes of AC in acutely ill medical patients. The diagnosis of AC was ascertained using the NEECHAM Confusion Scale. Of the 117 participants included in the larger study, 16 developed delirium (cumulative incidence estimate, 14%) and 10 died within 1 year of the index hospitalization. These 10 cases were categorized in two groups: those with a cancer-related diagnosis (n = 6) and those without cancer (n = 4). To further describe the nature of AC near the end of life, two case studies are presented. Because all previous studies were conducted using samples consisting of patients with cancer, it is unknown whether the findings reported in previous studies hold for other terminal illnesses, such as chronic obstructive pulmonary disease or heart failure. The data presented in this article suggest there are differences in baseline vulnerability (e.g., cognitive status) and the timing of AC in relation to death. These differences need to be explored in a larger sample of individuals both with and without a diagnosis of cancer. The severity and course of AC in the terminally ill population needs to be described to gain a better understanding of end-of-life AC phenomenology (e.g., signs, patterns, subtypes). Armed with this information, health care providers will then be able to develop and test AC specific treatments of patients, as well as counsel and support family members of patients experiencing AC. PMID- 11915157 TI - Achieving restraint-free care of acutely confused older adults. AB - Restraint-free care has emerged as an indicator of quality care for older adults in all settings. The most difficult challenges to achieving this goal are care of hospitalized older adults who are functionally dependent and cognitively impaired. The purpose of this article is to report findings from a descriptive study of restrained hip fracture patients, and discuss approaches to achieving restraint-free care. Rate of restraint use was 33.2% among hospitalized hip fracture patients during an 11-year period in 20 metropolitan teaching hospitals. Restrained patients were older men who resided in nursing homes prior to hospitalization. Clinically, restrained patients had a diagnosis of dementia, were noted to be confused or disoriented by nursing staff, and were dependent in activities of daily living. An individualized approach to care is the best method to avoid use of physical restraints for patients with acute confusion and cognitive impairment. PMID- 11915158 TI - A surprising moment. PMID- 11915159 TI - Gerontological nursing research: challenges for the new millennium. PMID- 11915160 TI - Leapfrog Group jumps at chance to give consumers health care info. AB - If you wanted to buy a home appliance, you could find more information than if you needed to choose the best hospital--until now. The Leapfrog Group has just released its first comparative hospital data for more than 200 hospitals nationwide. PMID- 11915161 TI - NRMI (National Registry of Myocardial Infarction) data impacts care on many different levels. PMID- 11915163 TI - A schedule change brings peace to rural Montana OR. PMID- 11915164 TI - See an MD in 30 minutes or your money back. PMID- 11915165 TI - Surgical management of breast cancer: today and tomorrow. AB - The surgeon continues to play a key role in the initial local/regional management of breast cancer. Breast conserving therapy (BCT) utilizing segmental mastectomy has been developed as an alternative to total mastectomy to improve cosmetic and psychosocial outcomes while retaining oncologic benefits. Developments in the surgical treatment of breast cancer have changed the way breast cancer is diagnosed and treated, and will likely continue to evolve in the future. This review highlights aspects of breast cancer treatment in which the surgical approach has evolved or is evolving, and comments on how the surgeon may participate in breast cancer care in the future. PMID- 11915166 TI - Practical dosimetry of 131I in patients with thyroid carcinoma. AB - Radioiodine treatments of patients with well-differentiated thyroid carcinoma have generally been safe and beneficial. Safety can be ensured while efficacy is increased through practical methods of dosimetry that measure body retention of 131I. Prescriptions for therapeutic 131I can be decreased when the retention level is high and increased when the level is low. Assays of serum free T4 will alert the physician to possible increased radiation to blood and bone marrow, and appreciable concentrations of free T4 are indications to reduce the therapeutic 131I. Carcinomas > or = 1 cm in diameter that are not visible on diagnostic scintigraphy are unlikely to respond to the commonly prescribed mCi of 131I. Biologic responses to commonly prescribed levels of therapeutic 131I, as seen in toxic changes of normal tissues and in indices of tumor size, will be the final dosimeters. With lower levels of prescribed diagnostic 131I, stunning should not impair dosimetry. Thus, readily obtained measurements make dosimetry a practical method for improving carcinoma therapy with 131I. PMID- 11915167 TI - Radiation dosimetry for radionuclide therapy in a nonmyeloablative strategy. AB - Radionuclide therapy extends the usefulness of radiation from localized disease of multifocal disease by combining radionuclides with disease-seeking drugs, such as antibodies or custom-designed synthetic agents. Like conventional radiotherapy, the effectiveness of targeted radionuclides is ultimately limited by the amount of undesired radiation given to a critical, dose-limiting normal tissue, most often the bone marrow. Because radionuclide therapy relies on biological delivery of radiation, its optimization and characterization are necessarily different than for conventional radiation therapy. However, the principals of radiobiology and of absorbed radiation dose remain important for predicting radiation effects. Fortunately, most radionuclides emit gamma rays that allow the measurement of isotope concentrations in both tumor and normal tissues in the body. By administering a small "test dose" of the intended therapeutic drug, the clinician can predict the radiation dose distribution in the patient. This can serve as a basis to predict therapy effectiveness, optimize drug selection, and select the appropriate drug dose, in order to provide the safest, most effective treatment for each patient. Although treatment planning for individual patients based upon tracer radiation dosimetry is an attractive concept and opportunity, practical considerations may dictate simpler solutions under some circumstances. There is agreement that radiation dosimetry (radiation absorbed dose distribution, cGy) should be utilized to establish the safety of a specific radionuclide drug during drug development, but it is less generally accepted that absorbed radiation dose should be used to determine the dose of radionuclide (radioactivity, GBq) to be administered to a specific patient (i.e., radiation dose-based therapy). However, radiation dosimetry can always be utilized as a tool for developing drugs, assessing clinical results, and establishing the safety of a specific radionuclide drug. Bone marrow dosimetry continues to be a "work in progress." Blood-derived and/or body-derived marrow dosimetry may be acceptable under specific conditions but clearly do not account for marrow and skeletal targeting of radionuclide. Marrow dosimetry can be expected to improve significantly but no method for marrow dosimetry seems likely to account for decreased bone marrow reserve. PMID- 11915168 TI - Antitumor immune responses derived from transgenic expression of CD40 ligand in myeloma cells. AB - Tumor cells engineered to express immunogenes have been used for cancer vaccines to induce the antitumor immunity and study the antitumor immune mechanisms derived from the immunogene expression. In the present study, we engineered a mouse myeloma cell line J558 with a cloned CD40 ligand (CD40L) gene. We demonstrated that (i) the engineered J558/CD40L tumor cells expressing the CD40 ligand molecule lost their tumorigenicity in syngeneic mice, and (ii) the inoculation of J558/CD40L tumor cells further lead to the protective immunity against wild-type J558 tumors. In animal studies using T-cell subset depleted mice, we further showed that the primary rejection of J558/CD40L tumors did not require T cells, but was mainly mediated by NK cells, whereas the effector phase of the protective immunity is mediated by CD8+ T cells. In addition, our data, for the first time, showed that the inoculation of engineered J558/CD40L tumor cells is able to stimulate stronger activation of dendritic cells with enhanced expression of B7-1 and ICAM-1 molecules than the wild-type J558 tumor cells Taken together, we demonstrated the antitumor effect of engineered J558/CD40L tumor cells that is mediated by the activation of the host dendritic cells in vivo. Our data indicate that the introduction of co-stimulatory CD40 ligand molecule will be useful as a new strategy of immunogene therapy against tumors. PMID- 11915169 TI - Dosimetry in a myeloablative setting. AB - In clinical therapy trials using high dosages of systemically administered radioactivity to treat cancer, myeloablation may occur. This is either an effect of the circulating radioactivity labeled to antibodies exposing the bone marrow to radiation, or it may occur because malignant cells in the bone marrow are targeted. Bone marrow cells may be targeted through antigens expressed on cells in the bone marrow or because radioactivity is targeted to the skeleton. Assessment of radiation absorbed dose to the marrow may be useful for dose escalation or individualized patient treatment planning. With successful preservation of marrow function with autologous marrow or peripheral blood stem cell transplantation, other normal organs may also receive sufficient radiation to show toxicity. Accurate dose estimates to these organs is important for the design of future studies in order to minimize or avoid toxicity. This paper reviews internally administered high dose radiation therapy studies, and examines the radiation absorbed dose estimates reported from these studies. PMID- 11915170 TI - Mitogen therapy for biological warfare/terrorist attacks and viral hemorrhagic fever control. AB - Ken Alibek was for 17 years a leader in Biopreparat, the Soviet Union's top secret agency involved in developing and stockpiling the most lethal bacteria, viruses, and toxins in the history of mankind before he defected with his family to the United States in 1992. Very contrite when he discovered he had been misled to believe that his efforts had been essential to the survival of his homeland, Alibek has become active sounding an alarm about, among other things, thousands of unemployed Russian scientists who have been seeking survival by selling their destructive expertise to rouge states and bioterrorists. Working full time in devising protective measures that might help control the damaging effects of terrorist attacks, Alibek has placed strong emphasis on stimulating nonspecific immunities of victims mainly with interleukins and other cytokines. A more productive alternative would be giving mitogens such as PHA and PWM to reinforce vaccine and antibiotic actions, at the same time stimulating protective immune, myelopoietic, and lymphopoietic responses. A key objective would be to find an effective management for the dreaded viral hemorrhagic fevers. Using Ebola infection as an experimental model, Yang et al. have shown that PHA can block both the viral secretions that inhibit neutrophil immune responses and the viral transmembrane glycoprotein that facilitates damage of the human endothelial cells responsible for the lethal hemorrhagic manifestations. Normal serum glycoproteins have in the past been clearly shown to inhibit the functions of PHA, thereby increasing dosage requirements. Extrapolation of this interaction with serum glycoproteins suggests that PHA given intravenously in adequate dosage should readily be able to block the deleterious Ebola virus glycoprotein effects. Data in an extensive classification of the hemorrhagic fever viruses recently presented by Barry make it possible to predict that mitogen therapy should be effective for virtually all of the disorders included. Therapeutic trials should best start with intravenous administration of PHA since this is the mitogen about which most is known and the only one given to humans, although the nonagglutinating advantages of fraction i.v. of PHA should be evaluated as a replacement. Functioning in a different mode, PWM has the advantage of much greater potency, and can be given either intravenously or orally, since these appear to be equally effective routes of administration. The best means of properly integrating the use of these mitogens needs to be determined. PMID- 11915171 TI - Malignant melanoma of nasal cavity: case report and review of the literature. PMID- 11915172 TI - The experimental research of R-phycoerythrin subunits on cancer treatment: a new photosensitizer in PDT. AB - The mouse tumor cell S180 and human liver carcinoma cell SMC 7721 cells were first treated with R-PE and its subunits (alpha, beta, gamma subunits), then irradiated with Argon laser (496 nm, 28.8 J/cm2). Survival rate was measured by MTT method. In order to compare the phototoxicity in normal cells, the mouse marrow cells were treated with photofrin II and beta-subunit, irradiated with 45 J/cm2 of light; survival rate was also measured by MTT method. The result showed that R-PE subunits had better PDT effect on s180 cells than R-PE and lower phototoxicity in marrow cells than photofrin II. Flow cytometric analysis showed that PDT results in a growth inhibition and a G0-G1 cell cycle arrest in SMC 7721 cells. The tumor cells inhibited by PDT in vivo were morphologically observed by TEM, the tumor cell death was due to the occlusion of tumor blood vessels and inducement of cell programmed death in nuclei. Therefore, with the advantage in special fluorescence activity, low molecular weight, good light absorbent character and weak phototoxicity, R-PE subunit is an attractive option for improving the selectivity of PDT. PMID- 11915173 TI - Peripheral blood immunological parameters for use as markers of pre-invasive to invasive colorectal cancer. AB - In cancer, the extent to which the disease has spread is probably the most important factor in determining patient prognosis. Hence practical and non invasive methods are needed to identify disease stage. In a previous paper we showed how diagnostic and prognostic indices for disease progression could be defined by evaluating parameters in peripheral blood. The aim of this study was to identify further serum parameters that could be used. Serum levels of interferon (IFN) gamma, interleukin (IL)4, IL8, IL7, IL1 beta, tumor necrosis factor (TNF) alpha, granulocyte macrophage-colony stimulating factor (GM-CSF), soluble (s) IL2 receptor (R) and sIL6R were studied but only levels of IL4, sIL2R, IL8 and IL7 were found to be significant and would therefore be of use in defining diagnostic and prognostic indices for disease progression. In further detail, our results indicate that when serum levels of sIL2R < 522 U/ml, IL4 < 159 pg/ml and IL8 > 339 pg/ml there is a 95% probability that the disease is in stage I or II where there is no infiltration of lymph nodes; when serum levels of sIL2R > or = 522 Ug/ml, 159 pg/ml < or = IL4 < or = 319 pg/ml, and IL7 < 54 pg/ml, there is a 95% probability that the disease is in stage III and the tumor has invaded the lymph nodes; when the serum levels of IL4 > or = 431 pg/ml and IL7 > or = 54 pg/ml, there is a 95% probability that the disease is in stage IV and there is metastasis. PMID- 11915174 TI - Irradiated cells from autologous tumor cell lines as patient-specific vaccine therapy in 125 patients with metastatic cancer: induction of delayed-type hypersensitivity to autologous tumor is associated with improved survival. AB - OBJECTIVE: We established short-term cultures of pure tumor cells for use as autologous tumor cell vaccines in an effort to study the effects of patients specific immunotherapy. PATIENTS AND METHODS: Surgically resected fresh tumor was obtained from patients with metastatic cancer. Successful tumor cell lines (5 x 10(7)) were expanded to 10(8) cells, irradiated, and cryopreserved for clinical use. Following a baseline test of delayed-type hypersensitivity (DTH) to an i.d. injection of 10(6) irradiated autologous tumor cells, patients received 3 weekly s.c. injections of 10(7) cells, had a repeat DTH test at week-4, then received monthly vaccinations for 5 months. A positive DTH test was defined as > or = 10 mm induration; survival was determined from the first DTH test. RESULTS: Short term cell lines were successfully established for 299/695 patients (43%). Vaccines were prepared for 231 patients, 142 of whom were treated, and 125 had a baseline DTH test recorded. Median follow up at the time of analysis was greater than 5 years. There was no difference in survival for any of the following: gender, age > 50 years, melanoma histology, anergy to common recall antigens or baseline DTH test result. Only 17 patients had a positive DTH at baseline (14%), but DTH converted from negative to positive in 31/80 (39%) of those who were tested, and in 31/108 (29%) of all patients (intent-to-convert analysis). For the 48 patients who were DTH-positive at entry, or converted to DTH-positive, the median survival was 30.5 months and 5-year survival 41% compared to 11.4 months and 9% 5-year survival for 77 patients whose DTH was never positive (P2 = 0.003). However, survival was even better for patients whose DTH test converted to positive compared to patients who were DTH-positive at baseline (median 37.5 vs 11.9 mos, P2 = 0.066). CONCLUSION: This patient-specific, cell culture-derived, autologous tumor cell vaccine induced anti-tumor immune reactivity that was associated with improved survival in patients with advanced cancer. PMID- 11915175 TI - Options for radionuclide therapy: from fixed activity to patient-specific treatment planning. AB - The therapeutic use of radioisotopes in medicine as unsealed sources has a long history dating back to the 1930s. The established and continuing objectives are to provide radiation dose to the target tissue at the desired cytotoxic level while avoiding or minimizing toxic effects. Selected radionuclide therapy protocols including 32P for polycythemia vera, 131I for Graves' disease, and 131I for postsurgical ablation of thyroid remnants in the management of differentiated thyroid cancer are presented for historical review with the focus on protocols for administering the radiopharmaceuticals and the role played by dosimetry. The discussion also includes consideration of complications and the assessment of outcome for these diseases. The vista for radionuclide therapy today is reviewed along with the options for determining the administered activity. Patient specific dosimetry encompasses a number of levels ranging from basic measurement of relevant biokinetic parameters and use of standard models to calculate (and extrapolate) radiation dose to sophisticated three-dimensional techniques employing fusion of physiologic and high-resolution anatomic images coupled with advanced 3-D voxel patient representation and Monte Carlo techniques for use in radiation dose calculation. The role of patient specific dosimetry in clinical trials (Phase I, II, III trials) along with its utility in treatment planning, follow-up evaluation, and elucidation of dose-response relationships is discussed. The challenge ahead for those who advocate patient specific dosimetry is to assemble the outcome data and perform the analysis to support this contention. PMID- 11915176 TI - Clinical trial design and scoring of radionuclide therapy endpoints: normal organ toxicity and tumor response. AB - Like other cancer therapy agents under development, radionuclide therapies are usually evaluated in a progressive series of clinical trials after basic science, human cell culture and animal model studies. Toxicities during these trials are graded using common scoring systems that are in widespread use such as the Common Toxicity Criteria from the National Cancer Institute. Information on normal tissue toxicity from radionuclides is more limited than that from external beam radiation and is more variable. Variability is likely due to many biologic factors as well as less precise dose quantitation than those used in external beam radiation practice. As expected based on known radiobiologic effects, tolerance to radionuclide therapy appears to exceed that from high dose rate external beam radiation in most organs. Although the correlation between reported dose estimates and toxicity has progressively and substantially improved over the past two decades, further progress is needed to establish optimal toxicity predictive relationships. Continued refinement of dosimetry techniques and standardization is expected to increase the accuracy and comparability of radiation dose reports between institutions as well as improve dose/response correlation. PMID- 11915177 TI - Stress proteins in cardiovascular research. AB - This review article is dealing with the most important aspects of stress proteins expression in cardiovascular research focused mainly on myocardial ischaemia and reperfusion. The major role of heat shock protein 70 kDa in endogenous route of myocardial protection is widely discussed in the context of possible contribution of other stress proteins. On the basis of numeral classical and recent published scientific data the problem of cellular response to stress and its possible clinical implication are presented. PMID- 11915178 TI - A special report I. Prion protein (Prp)--amyloid plaques in the transmissible spongiform encephalopathies (TSEs) or prion disease revisited. AB - We present a retrospective analysis of PrP-amyloid plaques encountered in CJD and GSS. In human TSEs (kuru, CJD and GSS) several PrP-immunopositive plaques and plaque-like deposits were detected. In kuru, plaques were typical "kuru" plaques- stellate structures deposited mostly in the granular and Purkinje cell layer of the cerebellum. Many smaller or larger clusters were visible but, in contrast to GSS, they never have merged together to form multicentric plaques. In all cases of GSS, plaques were located in the granular and Purkinje cell layer and in the molecular layer. There were many different forms of plaques: from kuru plaques (unicentric stellate plaques) to clusters of unicentric plaques which by merging eventually formed "multicentric plaques". The latter are the hallmark of this disease. By electron microscopy several types of amyloid plaques, which corresponded to those seen by PrP immunohistochemistry were observed. The first type, unicentric kuru plaque consisted of stellate arrangements (stars or cores) of amyloid bundles emanating from a densely interwoven center. Amyloid stars were surrounded by astrocytic processes and invaded by microglial cells but dystrophic neurites were only rarely seen. In contrast multicentric plaques were often surrounded by dystrophic neurites. The rarest type of plaque, were neuritic plaques. In 263K and 22C-H scrapie-infected hamster brains, on the light microscopy of the semi-thin (1 micron) sections, discrete PrP-immunopositive plaques were observed in the subependymal region but not in the deep brain neuroparenchyma. These plaques were not discernible by routine HE staining. Ultrastructurally, plaques were recognized as areas of low electron density containing haphazardly-oriented fibrils and not as stellate compact structures typical of plaques in human cases of CJD and GSS. These plaques were located beneath the basal border of the ependymal cells and adjacent blood vessels. Occasional dystrophic neurites containing electron-dense inclusion bodies were seen within the plaque perimeter, which always remained PrP-negative. PMID- 11915179 TI - The expression of vimentin in HL-60 cells induced with etoposide using immunofluorescence and immunogold methods. AB - HL-60 leukemic cells were treated with 6 different doses of etoposide for 72 hours. Changes in the distribution of vimentin were found to be dependent on the concentration of etoposide. As compared with control cells there were distinct changes in cells incubated with 100 and 200 microM/L of the drug. The size of cells treated with 100 microM/L and especially with 200 microM/L increased, but the number of cells decreased. In control cells and those treated with 0.02, 0.2 and 2 microM/L etoposide, vimentin was seen rather as a ring often with the increased concentration near one pole of the cells. Cells at 20 microM/L etoposide showed the same staining pattern but more brighter cells were found. The addition of 100 microM/L and 200 microM/L etoposide to cells resulted in diffusely distributed fluorescence staining, which often appeared as a quite dense network around the nucleus. Immunogold labelling was observed in cells treated with all doses of etoposide and control cells. Labelling was localized in the nucleus but also in the cytoplasm but rather in the area of the nucleus. PMID- 11915180 TI - Karyometric features differentiate early invasive cervical squamous cell carcinoma from preinvasive carcinoma. AB - The aim of the study was to investigate whether karyometric features reflect phenotypic change from preinvasive to early invasive squamous cell carcinoma of the cervix uteri. The material for the study was obtained from 12 patients diagnosed in the Chair of Pathomorphology. Each specimen contained both preinvasive and microinvasive lesions. The system of image acquisition and analysis consisted of a microscope coupled with a CCD camera and a PC computer. The software based on AnalySIS 3.0 system (Soft Imaging Systems GmbH) automatically detected cell nuclei and measured the preselected parameters. In each case 250 cell nuceli from both preinvasive and invasive components were analyzed. Significant differences in basic karyometric parameters were found both within individual cases and especially between groups studied. The present findings confirm the presence of early cytological changes at the onset of invasion. From the practical viewpoint the results suggest that karyometric features measured by image analysis might be used as a diagnostic tool, especially in cases where the diagnosis of microinvasion is difficult. PMID- 11915182 TI - Evaluation of the proliferative activity as an adjuvant helpful tool in the diagnosis of primary gastric lymphoma in gastrobiopsy. AB - Gastrobiopsy specimens obtained from 15 patients, in whom gastroscopy examination pointed to gastric cancer and primary gastric lymphoma was later diagnosed in surgical material after gastrectomy, were immunostained using the anti MIB-1 antibody to assess the proliferative index. A control study was carried out on representative surgical specimens from these patients and on 10 gastrobiopsy specimens from patients in whom histopathology revealed moderate/severe gastritis. The proliferative activity was assessed with respect to the type of lymphoma (primary gastric marginal zone B-cell lymphoma of mucosa-associated lymphoid tissue MALT type vs. diffuse large B-cell lymphoma) and stage of the lymphoma, and we compared the proliferative indices between inflammatory infiltrate and lymphoma cells. There were significant differences in the proliferative indices between inflammatory infiltrate and lymphoma cells, with immunohistochemical reactivity for MIB-1 antigen being visible even in the case of markedly mechanically damaged (crushed) gastrobiopsy material and accompanying necrosis. There were also differences in the proliferative indices between the groups of lymphomas varying in stage, although without statistical significance. There were no statistical differences in the rate of proliferation between lymphomas varying in "malignancy". The present results argue for the auxiliary role of the assessment of proliferative activity in gastrobiopsy material facilitating differentiation between inflammatory infiltrate and neoplastic infiltrate, especially in cases where mechanical damage or necrosis do not allow for precise histopathological evaluation. PMID- 11915181 TI - Fhit protein expression in endometrial cancers: no correlation with histological grade. AB - The genes specifically involved in endometrial cancers have not yet been discovered. The FHIT gene, a tumour suppressor located at 3p14.2, is altered in many human tumours, including those derived from the female genital tract. We have thus investigated the status of Fhit protein expression in endometrial carcinomas (EC), and its association with histological grade of malignancy in order to determine if Fhit expression is inactivated in EC and if so, whether it is inactivated during initiation or progression. Recent studies have reported that alteration in the FHIT locus detected by DNA and RNA analysis is well correlated with loss of Fhit protein expression in tumours. Thus, we characterised Fhit protein expression as an indication of FHIT gene status in 35 cases of EC of different histological grade (G1: 13 cases; G2: 14 cases; G3: 8 cases). In our group of cancers, Fhit protein expression was absent or reduced in 37% (13/35) of EC. The first 13 cases, judged as G1, showed Fhit deficiency in approximately 38.5% of cases (5/13). For G2 and G3 tumours these numbers were similar and accounted for approximately 35.7% (5/14) and approximately 37.5% (3/8), respectively. No statistical difference was found for Fhit expression among the various groups of tumours, which allowed us to conclude that morphological grade does not seem to be an important factor. Our results suggest that Fhit inactivation is an early event in carcinogenesis of the endometrium. As this observation is contrary to some already published reports, another independent study with larger amounts of material is necessary to determine this issue definitely. PMID- 11915184 TI - New educational methods in cytopathology: a transnational training program in cervical cytology--CYTOTRAIN CD-ROM. AB - Among the reasons for the low effectiveness of preventive measures for early detection of cancers and preinvasive states of the uterine cervix there is insufficient training of people performing cytological screening. In order to decrease the rate of false negative diagnosis it is necessary to improve the organisation of permanent professional training of cytopathologists and cytotechnicians. PMID- 11915183 TI - Quantitative analysis of interstitial mast cells in lupus and non-lupus membranous glomerulopathy. AB - Eleven renal biopsy specimens from patients with lupus membranous glomerulopathy (LMGN) and 16 from patients with primary (nonlupus) membranous glomerulopathy (NLMGN) for whom light, electron microscopy and immunofluorescence microscopy, and full clinical data were available were examined quantitatively. As a control 10 biopsy specimens of the kidneys removed because of trauma were used. Morphometric investigations were performed by means of a computer image analysis system to evaluate whether mast cells have a role in tubulointerstitial fibrosis in lupus and nonlupus membranous glomerulopathy and to examine the relationship between mast cells and interstitial alpha-smooth muscle actin (alpha-SMA) expression as well as interstitial infiltrates. The morphometric study revealed that the mean values of interstitial tryptase positive cells, expression of alpha SMA, interstitial volume, CD68+, CD45RB+, CD43+ and CD20+ cells were significantly increased in LMGN as compared with NLMGN. In both LMGN and NLMGN groups there were significant positive correlations between interstitial tryptase positive cells and interstitial expression of alpha-SMA, interstitial volume, serum creatinine as well as CD68+ cells. The present data suggest that in cases of membranous glomerulopathy with a large number of interstitial mast cells systemic lupus erythematosus should be taken into consideration, even if this aetiology was not clinically suggested at the time of biopsy. Additionally, in both LMGN and NLMGN significant positive correlations between interstitial mast cell count and relative interstitial volume support the role of these cells in the development of interstitial fibrosis, however this relationship needs further investigations. PMID- 11915185 TI - Giant cell myocarditis--a case report. AB - The authors described a case of giant cell myocarditis treated by heart transplantation. PMID- 11915186 TI - A technical note on microsatellite DNA instability studied in archival paraffin embedded tissues. AB - The authors present brief overview of problems associated with analysis of microsatellite sequences in DNA from archival paraffin-embedded tissue. This methodology can serve several diagnostic and research purposes provided appropriate precautions are taken to extract DNA of acceptable quality with small admixture of contaminants. Also PCR amplification step requires some modifications compared to routine reactions involving DNA from fresh or freshly frozen tissues. PMID- 11915188 TI - [Transnasal esophagogastroduodenoscopy?]. AB - INTRODUCTION: The most proper examination for the morphological changes of the upper gastrointestinal tract is endoscopy. The standard oro-gastric route is carried out under conscious sedation, but the drugs used might have cardiopulmonary side effects. Furthermore there are a few conditions when this routine endoscopy is technically impossible. AIMS: Could these failures be avoided with an endoscope led through the nose? Are there advantages of the use of ultrathin endoscopes for transnasal oesophago-gastro-duodenoscopy? PATIENTS/METHODS: Own experiences with transnasal therapeutic endoscopies on patients with cancer in the head and neck region compared with data from the literature. RESULTS: Based on literature and on own experiences, transnasal oesophago-gastro-duodenoscopy may be advised in cases with trismus, stenosis of the upper aero-digestive tract, oropharyngeal dysphagia, cricopharyngeal achalasia, in several neurosurgical conditions and on severely ill ventilated patients. I.v. premedication is not always necessary for transnasal oesophago gastro-duodenoscopy done with ultrathin endoscopes. CONCLUSIONS: The oesophagus might be accessed also via the nose, but it must be emphasized that transnasal oesophago-gastro-duodenoscopy is only advised when the standard way of upper gastrointestinal tract endoscopy is impossible. PMID- 11915189 TI - [Tumor-like tracheal and lung diseases in biopsy samples]. AB - Tumor like miscellaneous lung diseases are rare conditions. They can be confused with benign and malignant tumors. We describe the clinicopathological features some of this conditions i.e.: two cases of osteoplastic tracheopathy, one inflammatory tracheal polyp, two cases of tumor mimicking foreign body aspiration, and one case of tracheobronchial amyloidosis. PMID- 11915187 TI - [History of atrial fibrillation]. AB - The authors review the history of atrial fibrillation, the most frequent clinically observed cardiac arrhythmia. A French "clinicopathologist", Jean Baptist de Senac (1693-1770), was the first who assumed a correlation between "rebellious palpitation" and a stenosis of the mitral valve. From an analysis of simultaneously recorded arterial and venous pressure curves, the Scottish Sir James Mackenzie (1853-1925) demonstrated that a praesystolic a wave can not be seen on the jugular phlebogram during "pulsus irregularis perpetuus". The first human ECG depicting atrial fibrillation was published by Willem Einthoven (1860 1927) in 1906. The proof of a direct connection between absolute arrhythmia and auricular fibrillation was established by two Viennese physicians, Rothberger and Winterberg. The major discoveries relating to the pathomechanism and the clinical features of atrial fibrillation in the 20th century stemmed from the scientific activities of Karel Frederik Wenckebach, Sir Thomas Lewis, Gordon Moe and Maurits Allessie. PMID- 11915190 TI - [Heterotaxy syndrome, analysis of 13 cases and review of the literature]. AB - INTRODUCTION: Heterotaxy syndrome (Ivemark syndrome, or asplenia-polysplenia syndrome) is a heterogeneous group of disease with disturbed body symmetry and malposition of internal organs. Heterotaxy syndrome is caused by the disturbance of the left/right axis in the early embryonic period. AIM OF THE STUDY: The most frequency of heterotaxy syndrome's concomitant anomalies during a five year period in own fetopathology material. MATERIALS AND METHODS: Data of fetopathologic examination of 13 fetuses suffering from prenatally diagnosed heterotaxy syndrome. RESULTS: Situs ambiguus was detected in 9 cases out of 13. In the remaining 4 cases situs inversus totalis was diagnosed. The most frequent and important associated malformation included congenital heart disease was AV channel (10/13) and great vessel anomaly (10/13). CONCLUSION: In cases with prenatally detected complex cardiac anomalies (especially AV channel cases) heterotaxy anomaly must be taken into consideration, with main consequences in prenatal counselling. PMID- 11915191 TI - [Veresaev, the Russian physician writer]. PMID- 11915192 TI - [Removal of a bottle from the rectum. 1865]. PMID- 11915193 TI - Treatment of congenitally missing maxillary lateral incisors: orthodontics, bone grafts, and osseointegrated implants. PMID- 11915194 TI - Shirley Tilghman, PhD. PMID- 11915195 TI - Tobacco dependence treatment services in New Jersey. PMID- 11915196 TI - Mosquito Smith. PMID- 11915197 TI - Office-based claims--a growing concern. PMID- 11915198 TI - Joint negotiations legislation. PMID- 11915199 TI - Utah healthcare system watches over Olympians and spectators. AB - For the first time in Olympic history, a single organization was tapped to provide medical services when Intermountain Health Care (IHC), Salt Lake City, was named for the 2002 Olympic and Paralympic Games in Utah. IHC, a charitable, community owned, integrated, not-for-profit healthcare system serving patients in Utah and Idaho, spent four years developing and implementing a plan to deliver medical services to both Olympians and spectators. Nearly 350 IHC doctors, nurses, physical therapists and other professionals donated their services for the Olympics without compensation as part of their not-for-profit mission. In addition, about 1,000 IHC employees applied to be general volunteers during the games. PMID- 11915200 TI - Making the most of a wall calendar. Kingston Regional Health creates three for the price of one. AB - In a few short weeks, Kingston Regional Health, Kingston, N.Y., produced three versions of its 2002 wall calendar, one representing the health system itself, another its Women's Health Services subsidiary and the third, its Lifeline personal response and support service. PMID- 11915201 TI - You don't have to be an artist. Foundation for Hospital Art provides projects that work. AB - Wolfson's Children's Hospital in Jacksonville, Fla., and Cardinal Glennon Children's Hospital in St. Louis, were participants in PaintFest, a program by the Foundation for Hospital Art to provide colorful, soothing artwork for hospitals. More than 500 hospitals have participated since the Foundation began in 1984. Seasoned veterans express amazement at the overwhelming reaction by news media. PMID- 11915202 TI - A new kind of hospital magazine. Holy Name Hospital reaches Bergen County coffee tables. AB - Holy Name Hospital, Bergen County, N.J., mails its new magazine to 135,000 upscale homes, hoping to cause some defection from the heavy competition. The new publication retains the popular HealthWatch newsletter name, wrapping it in a slick package of consumer articles and ads. PMID- 11915203 TI - Cooley building opens in Houston. Demonstrates value of fully integrated marketing communications. AB - The Texas Heart Institute at St. Luke's Episcopal HospiTal in Houston dedicated its new 10-story Denton A. Cooley Building in January. The structure opened with a fanfare, thanks to a well-integrated marketing communications program. PMID- 11915204 TI - Web site helps families cope with childhood illnesses. Children's Hospital of Philadelphia undergoes Internet expansion. AB - The Children's Hospital of Philadelphia has received accolades for its revised Web site, which provides extensive communications channels for patients and their parents. Also, its intranet streamlines internal communications. PMID- 11915206 TI - Price survey. Suture price increases on the way. PMID- 11915205 TI - New employee strategies pay off. Identity crisis at Jeanes Hospital gives way to a new day. AB - A new spirit is evident at Jeanes Hospital in Philadelphia, where morale had slumped following the combination of the 128-year old Quaker-based institution with the Temple University Health System. The key is a new CEO with a passion for open communication, honest messaging and consistent follow-up. PMID- 11915207 TI - Ascension, Consorta craft new contract relationship. PMID- 11915208 TI - Hospitals to increase spending on membranes. PMID- 11915209 TI - Comeg turns repair calls into new product sales. PMID- 11915210 TI - No code not equal to no care. PMID- 11915211 TI - Becoming more culturally sensitive. PMID- 11915212 TI - Billing Medicare. Are you ready? PMID- 11915213 TI - Right on the money. Stimulate cash flow with software that suits your practice. PMID- 11915214 TI - Price point testing. How do we really know what to charge? PMID- 11915216 TI - BIOLASE receives first-ever FDA clearance for complete laser root canal therapy. PMID- 11915215 TI - FDA approves first biologic treatment for sepsis. PMID- 11915217 TI - ReD hot hygiene, Part 4. H is for helpful. PMID- 11915218 TI - Using flowable composite as a base for amalgam restorations. PMID- 11915220 TI - How to make exquisite cosmetic dentures a profitable part of your practice. PMID- 11915219 TI - Correction of an aesthetic problem related to excessive display of gingiva. PMID- 11915221 TI - Full-mouth reconstruction with increased vertical dimension. Case report. PMID- 11915222 TI - Another look at root canal obturation. PMID- 11915223 TI - Differential diagnosis of severe recurrent oral ulceration. PMID- 11915224 TI - The Class III posterior composite restoration. AB - This conservative method of restoring proximal carious lesions in posterior teeth is not offered as an "alternative" to more familiar methods. Rather, in the author's view, anyone who embraces the philosophy of "minimal invasion" and respects the preservation of sound tooth structure should first consider a class III composite resin restoration for aesthetic treatment of proximal caries in posterior teeth before using any other more aggressive or complex cavity design. In the practice of conservative dentistry, the author feels that the conventional class II approach is the less desirable alternative. PMID- 11915225 TI - Designing the dental office laboratory. PMID- 11915226 TI - Lifestyle modifications and the management of hypertension. PMID- 11915227 TI - Recession-proof your practice. PMID- 11915228 TI - Leadership. Half the battle. AB - A study of private and public sector managers, including 2,000 from the NHS, revealed consensus on the qualities of an effective leader. These included: concern for others, approachability, intellectual ability, communication skills, charisma and the ability to unite people. Integrity was seen as particularly important by public sector managers. Public sector managers were rated significantly higher by their staff than those in the private sector. The results point to the need for caution in introducing private sector managers to the NHS. PMID- 11915229 TI - Primary care. Take it from here. AB - Putting six advice workers into general practices for three hours a week over 10 months resulted in 500,000 Pounds being identified in benefits for patients. Many referrals came from GPs, but patients referred themselves in a fifth of cases. Patients welcomed the service and found it convenient. The co-operation of reception staff was vital. PMID- 11915230 TI - Long-term care. The best of both worlds. AB - An NHS/housing association venture has produced good accommodation for 30 elderly people in need of long-term care. The housing organisation takes responsibility for support services such as laundry, cleaning, security and catering. The unit remains part of the NHS. Patients appreciate the environment and staff turnover is low. PMID- 11915232 TI - Noise attenuation characteristics of a foam type earplug measured using a dummy head. AB - The attenuation characteristics of the earplug using a dummy head (KEMAR; Knowles Electronics Manikin for Acoustic Research) were measured, when it was exposed to pure tones or broadband noises at three different sound pressure levels (SPLs) of 70, 80 and 90 dB SPL in the 'wide frequency range from 250 to 20 k Hz. We ascertained the satisfactory capacity of noise attenuation except for from 10 k to 14 k Hz. The noise attenuation level increased with frequency until 2 k Hz, became almost constant until 8 k Hz, then decreased at around 10 k to 14 k Hz, and recovered in the range from 16 k to 20 k Hz. The frequency characteristics were identical among three different sound pressure levels and almost identical between pure tones and broadband noises. The change of frequency characteristics affected by the depth of the earplug that was inserted into the artificial ear canal of the KEMAR was also investigated. Measured sound pressure level by the builetin microphone in the KEMAR revealed a region in which the measured sound pressure levels were relatively higher around 10 k Hz. The frequency of the region moved higher when the earplug was inserted more deeply. Therefore it was considered that this phenomenon was related to the resonance frequency of the cylinder-like cavity made between the end section of the earplug and the diaphragm of the built-in microphone. PMID- 11915231 TI - Effect of endocrine disrupters on immune responses in vitro. AB - It has been reported that endocrine disrupters (EDs) have several biological activities, especially on endocrine systems. We studied the effect of EDs such as diethyl stilbesterol (DES), 4,4'-isopropylidene diphenol (BPA), bis (2 ethylhexyl) phthalate (EHP) and p-nonylphenol (NP) on murine spleen cells in vitro. DES, BPA, EHP and NP enhanced concanavalin A- and lipopolysaccharide induced proliferative responses of spleen cells. The activity was most effective at the concentration of 10(-7) M. These EDs stimulated both T cells and B cells. They also enhanced cytokine productions such as interleukin (IL)-4 and interferon gamma, and the total immunoglobulin G production of spleen cells. Furthermore, these EDs enhanced cytokine productions such as tumor necrosis factor and IL-1 by macrophages. These results suggest that EDs have a modulating activity on immune responses. PMID- 11915233 TI - An overview of occupational health and safety in Australia. AB - Australia is a developed country with a high standard of living, small population and large land area. Manufacturing is currently the largest economic contributor, although mining and agriculture are also significant industries. There are around 10 million employees in total, with retail trade and manufacturing being the largest employers. Manufacturing currently has the highest incidence of workplace injury, although around 5% of all Australian workers suffer from some kind of occupational disease or injury every year. Occupational Health and Safety (OHS) legislation is individually managed and enforced by the 8 states and territories. Training and registration for OHS professionals varies between the speciaities and usually requires a combination of academic qualifications and workplace experience. Non-medical personnel constitute a large proportion of OHS professionals in Australia. PMID- 11915234 TI - [Chemokines in health and diseases]. AB - A large number of lymphocytes are recruited to the sites of inflammation where they form an essential component for the response to infection, injury, autoimmune disorders, allergy, tumor invasion, atherosclerosis and so on. The recruitment of leukocytes into tissue is regulated by a sequence of interactions between the circulating leukocytes and the endothelial cells. Leukocyte integrins play a pivotal role in leukocyte adhesion to endothelial cells. During the process, the activation of integrins by chemokines is essential for integrin mediated adhesion in which a signal transduced to the leukocyte converts the functionally inactive integrin to an active adhesive configuration. Such a regulation is most emphasized in inflammatory processes, in which leukocytes migrate from circulation into the tissue. The concept, concerning chemokines in diseases and health, would be an enormous help in clarifying pathological processes of inflammation as well as the new pharmacological approaches to more specifically control particular inflammation. PMID- 11915235 TI - [Gait analysis of idiopathic normal pressure hydrocephalus]. AB - Normal pressure hydrocephalus (NPH) is a clinical syndrome associated with dementia, gait disturbance and urinary incontinence. Gait disturbance is usually the initial sign and most important symptom, but its objective evaluation has not been established. We analyzed the gait of an idiopathic NPH before and after ventricular shunting with the gait analysis system. Before shunting, the stride was short and irregular, and the truncal movement was unsteady. Three-dimensional patterns of angular relationships between 3 joints, namely the ankle, knee and hip were small and irregular. The vector profile of floor reaction force showed a monophasic pattern with absence of the peak at toe-off. After shunting, the step enlarged and the truncal movement was steady. The three-dimensional patterns of angular relationships between the 3 joints were nearly normalized. The vector profile of floor reaction force showed an appearance of the peak at toe-off, which formed a biphasic pattern, similar to the pattern of a normal person. The gait analysis is a useful method to evaluate gait disturbance in idiopathic NPH. PMID- 11915236 TI - [Two cases of Swyer-James syndrome]. AB - Two cases of Swyer-James syndrome are reported. Case 1; A 25-year-old man was admitted to our hospital to be treated for Mycobacterium avium infection. Chest X ray film revealed hyperlucency in the right lower lung. High-resolution CT showed a low attenuation area and bronchiectasis in the right lower lobe. He had had two episodes of pneumonia in his childhood. 3D CT scan showed a narrowing of right lower pulmonary arteries. Case 2; A 65-year-old woman was admitted to our hospital with dyspnea on effort. Chest X-ray film revealed hyperlucency in the right lung. Chest CT scan on inspiration and expiration detected air-trapping, which is characteristic of this syndrome. In both cases, the volume of the hyper lucent lung was normal. From these clinical findings, we diagnosed these two cases as Swyer-James syndrome and in this paper described the clinical features and treatment. PMID- 11915237 TI - [Four cases of non-clostridial gas gangrene with diabetes mellitus]. AB - We report four cases of non-clostridial gas gangrene. All cases were associated with diabetes mellitus as the underlying disease. Case 1: a 60-year-old male developed an ulcerative lesion on the dorsum of his left foot. Peptostreptococcus asaccharolyticus, Citrobacter freundii and Staphyrococcus epidermidis were identified in culture from odoriferous pus. Case 2: a 81-year-old female developed a lesion on her vulva spreading to the right lower abdomen. Bacteroides bivius, Peptostreptococcus asaccharolyticus and Streptococcus faecalis were identified in culture of the odoriferous pus. Case 3: a 80-year-old male developed a swollen area with ulcer on the right foot. Bacteroides fragiris, Enterococcus faecalis, Proteus mirabilis, Enterococcus avium and Enterococcus faecalis were identified by culture. Case 4: a 52-year-old female developed swelling of her left groin. Enterococcus faecalis and Streptococcus anginosus were identified in culture from the odoriferous pus. In all patients, a radiological examination revealed the presence of subcutaneous gas in the lesion. Prognosis of non-clostridial gas gangrene is usually poor. These four patients, however, all survived. Once an infectious sign is seen in the diabetic patient, it is important to discover a gas figure by using the radiological examination (plain film or computed tomography). Earlier diagnosis and debridement are the most important for a better prognosis. Because workers with diabetes mellitus are now increasing in number, occupational physicians should always keep in mind that a serious infectious disease like non-clostridial gas gangrene can develop even from minor accidental trauma, and they should control the working environment in the workplace where accidents often happen. PMID- 11915240 TI - [Report from the 16th UOEH Meeting of Gastrointestinal Image Diagnosis]. PMID- 11915239 TI - [Acute effects of methyl bromide gas in rats and mice--studies on survival time]. AB - Wistar male rats and ddY male mice were exposed to 500-10,000 ppm of methyl bromide gas up to 8 hours, and the survival time and weights of brain, lung, liver, kidneys, spleen and tests were measured. The survival time decreased with the gas concentration almost exponentially. In general, rats had a longer survival time than mice under the same exposure concentration. Difference of survival time between rats and mice was remarkable at low concentrations but small at high concentrations. Spleen weight decreased significantly and kidney weights tended to increase at 2000 ppm or greater exposure concentrations. Difference between wet and dry lung weight in the exposure group was significantly greater than that in the control group, especially for high concentrations, which suggested pulmonary edema or bronchopneumonia. PMID- 11915238 TI - [Are employees who have retired due to occupational maladjustment adaptable to new workplaces?--Character traits as the individual factors]. AB - Determining factors for adaptation to a new workplace in light of 3 cases that have taken early retirement due to occupational maladjustment are discussed. The character traits of these 3 cases are regarded as individual factors because of their conspicuousness. In regard to adaptation to new workplaces, two of my cases were unable to adapt in their previous workplaces because of their avoidant personality. They were also unable to adjust to their present workplaces. When the worker's character traits tend to cause occupational maladjustment, appropriate approaches including counseling and occupational adjustment should be recommended to the worker. PMID- 11915241 TI - A gift of song. PMID- 11915242 TI - Spiritual care. Safe, appropriate, ethical. PMID- 11915243 TI - How can we pray for nursing? PMID- 11915244 TI - Dark night or spiritual peace? Lessons from a 16th century mystic. PMID- 11915245 TI - Elizabeth's little black box. PMID- 11915246 TI - Is spiritual care ethical? PMID- 11915247 TI - No DNR. A time to walk? PMID- 11915248 TI - Ministry in ICU. PMID- 11915249 TI - Nutu's leg. PMID- 11915250 TI - Summer in the metro jail. PMID- 11915251 TI - Why I love nursing: directed steps. PMID- 11915252 TI - Embracing shalom. Moving into fullness of life. PMID- 11915253 TI - Journey of a lamb. Walking through dementia together. PMID- 11915254 TI - Subjective responses to caregiving for a spouse with dementia. AB - Family members, typically spouses, serve as the primary caregivers for individuals with dementia. While it is recognized that caregivers experience both positive and negative subjective responses to caregiving, relatively little research has been done on the relationship between these types of responses. The objectives of the study were to examine the relationships between and among spouse caregivers' positive and negative subjective responses to caregiving, and increase understanding of the experience of being a spouse caregiver for a person with dementia. Fifty spouse caregivers completed quantitative measures of positive subjective responses (Positive Aspects of Caregiving and Caregiver Competence) and negative subjective responses (Relational Deprivation, Role Captivity, and Loss of Self). Additionally, participants were interviewed concerning their caregiving experiences. Positive Aspects of Caregiving and Caregiving Competence were significantly related to each other (p < .01), as were the three measures of negative subjective responses (p < .01). No significant correlations were found between any measures of a positive and a negative subjective response, suggesting the two types of responses are independent. Duration of caregiving was significantly related to Positive Aspects of Caregiving (p < .05), Caregiver Competence (p < .05), and Relational Deprivation (p < .01). Qualitative interview data revealed that participants simultaneously experienced caregiving as self-affirming, while also enduring losses and difficulties resulting from their caregiving role. Integrating the positive and negative aspects of the whole of caregiving is important to understand the caregiving experience and to design interventions to support caregivers. PMID- 11915255 TI - Modeling nursing scientific inquiry. The University of Iowa Hospitals and Clinics Research Utilization Residency Program for Geriatric Nurse Practitioners and Gerontological Clinical Nurse Specialists. AB - The application of research findings to achieve nursing outcomes sensitive to the effects of nursing care remains a challenge for the nursing profession. The Research Utilization Nursing Residency Program is a project implemented by the University of Iowa Gerontological Nursing Interventions Research Center. For a discharge planning coordinator serving a largely geriatric patient population, the residency experience provided support to apply scientific nursing knowledge to outcomes. Resources from both clinical and academic settings are discussed within the context of the resident's experience. PMID- 11915256 TI - Benefits of a nurse-managed wellness program. A senior center model. Using community-based sites for older adult intervention and self-care activities may promote an ability to maintain an independent lifestyle. AB - Recent health initiatives have encouraged the promotion of wellness and self-care among the elderly population. This research describes the effectiveness of a nurse-managed wellness program located at a multi-purpose senior center. The wellness clinic was created to provide a broad range of screening services, individual counseling services, health educational programs, and management services for chronic diseases common among older adults. Outcome measures focusing for utilization of health services, healthy behaviors, emotional well being, and mental and physical health variables gathered from 111 older adults demonstrated significant program success. Respondents who used the wellness clinic more frequently, and in particular, those who consulted regularly with the clinical nurse, were more likely to increase their healthy behavior and subsequent health knowledge. As a result of participation in the wellness programs, respondents reported a greater psychological comfort and a more confident feeling concerning the ability to maintain an independent lifestyle. PMID- 11915258 TI - Lore K. Wright, PhD, RN, CS, FAAN 1938 to 2001. PMID- 11915257 TI - Pain in nursing home residents. Comparison of residents' self-report and nursing assistants' perceptions. Incongruencies exist in resident and caregiver reports of pain; therefore, pain management education is needed to prevent suffering. AB - Physical pain is a significant problem for many older adults, and as many as 83% of nursing home residents reportedly have pain. Unrelieved pain has consequences for elderly individuals' physical and mental health, rehabilitation, and quality of life. Evidence suggests, however, that pain is underdetected and poorly managed among older adults. This may be due, in part, to lack of congruence between patients' and caregivers' perceptions of pain. Thus, the purpose of this study was to investigate: the prevalence, location, and intensity of residents' self-rated and nursing assistants' (NA) rated pain; the congruence between residents' and NAs' ratings of pain; and resident-based and NA-based correlates of congruent and incongruent pain ratings. Participants in this study were 45 nursing home resident-NA dyads. The results indicated 49% of residents stated they experienced pain in the past week, but NAs reported that 36% of residents experienced pain during the same time interval. There was no significant association between residents' self-ratings and NAs' ratings of pain. Of the 45 paired ratings, residents and NAs were congruent in 37.7% of cases and incongruent in 62.2% of cases. Incongruent ratings included both underdetection (37.8%) and overreporting (24.4%) by the NAs. Only residents' self-rated affect (e.g., depression, well-being) was significantly associated with whether their pain was congruently assessed, underdetected, or overreported. Depression was highest in those for whom pain was not perceived by the NAs and well-being was highest in those residents who denied pain but for whom NAs reported pain. Caregiver characteristics (e.g., age, education, work experience) were not significantly associated with pain congruence outcomes. These findings illustrate the complexities of assessing pain in older adults, and the need to include nursing assistants (NAs) in educational programs focusing on managing pain in elderly nursing home residents. PMID- 11915259 TI - The week of crying at work. PMID- 11915260 TI - Measuring quality of care with an inpatient elderly population. The geriatric resource nurse model. AB - Nurses provide health services to an increasing number of older adults in acute care settings. Acute care nurses are committed to giving patients the highest quality care while recognizing the importance of delivering care in a cost effective manner. In this study, a unit-based, nurse-centered geriatric program is evaluated. The program is designed to enhance the knowledge and skill of staff nurses in providing care to elderly patients. Both quantitative and qualitative methods are used to assess geriatric resource nurses' (GRNs) influence on quality and cost outcomes of the elderly participants. Patients age 65 years and older were randomly selected from two general medical units of a major academic tertiary care center in the southeastern United States. Data were collected during an 18-month period in 1996 and 1997. A total of 129 participants provided data for quantitative analysis. A subset of 34 participants (17 from the unit where GRNs were on staff and 17 from a control unit) was interviewed about their experience during hospitalization. This information was analyzed for common themes and trends using appropriate qualitative techniques. Demographic variables and common measures of illness severity and complexity showed comparable patient populations on the two units. However, results of quantitative analyses indicated significant differences between groups on admission for several of the health status measures. Participants on the unit without GRNs were found to have more problems with pain, incontinence, and mobility. Administrative measures showed the number of patients readmitted to the hospital within 31 days of discharge and the length of stay associated with this initial readmission were significantly lower on the unit with GRNs. The use of vest-type physical restraints was also less frequent on this unit. Elderly patients in both groups indicated they have special needs related to normal aging changes and chronic illnesses, resulting in higher levels of fragility and decreased energy reserves. They identified specific functional areas for which help was needed. These include assistance with bathing, eating, sleeping, mobility, and elimination. Fewer participants on the intervention unit reported decline in activities of daily living (ADL) function during hospitalization than did control participants. Participants in both groups stressed the importance of nurses' demonstrating understanding and caring when working with older individuals. PMID- 11915261 TI - Mary Starke Harper. "I love doing the impossible". PMID- 11915262 TI - Research-based protocol. Management of constipation. PMID- 11915263 TI - MSHAKE. A tool for measuring staff knowledge related to geriatric mental health. AB - This article describes the development of a knowledge-screening instrument used with staff caring for elderly patients with neuropsychiatric disability. A 25 item tool, referred to as the Mary Starke Harper Aging Knowledge Exam (MSHAKE), was developed to verify knowledge competencies of 171 employees at a geropsychiatric center in the Southeastern United States. The MSHAKE assesses basic geriatric mental health knowledge in an educationally diverse health care work force. A variety of descriptive and inferential statistics provide preliminary evidence supporting the use of the MSHAKE as an effective measure of staff members' essential knowledge of aging and neuropsychiatric disorders. This tool has practical application in the clinical setting as part of a competency verification system or in-service evaluation. PMID- 11915265 TI - Nursing home placement. Factors affecting caregivers' decisions to place family members with dementia. AB - Nurses who work with older adults with dementia in the community are aware of how difficult it can be for family members to make placement-related decisions, and need research-based information to help families make these decisions. This article presents the results of a qualitative research study intended to explore the factors influencing caregivers' decision-making process related to placing an elderly family member with dementia in a nursing home. Using the study findings, the author suggests a theory which helps to explain some of the decision-making factors which appear to influence caregivers' abilities to make placement-related decisions. The nursing implications of this theory, and recommendations for future research, are presented. PMID- 11915264 TI - Long-term care for older adults. The role of the family. AB - Family members are involved in all aspects of a frail or ill older adult's care before, during, and after admission into a long-term care facility. However, their role within the facility is not always clearly defined. This article addresses issues regarding facility policies and staff attitudes toward family involvement, particularly whether the family member is seen as an integral part of the care system or peripheral to the care system. An emphasis is placed on the use of open communication to prevent misunderstandings and disagreements between staff and family and to promote good quality care and acceptable quality of life for the resident. PMID- 11915266 TI - Aging is like farming. PMID- 11915267 TI - Half-heimer's disease? PMID- 11915268 TI - Quality in end-of-life care. PMID- 11915269 TI - Easy rider wheelchair biking. A nursing-recreation therapy clinical trial for the treatment of depression. AB - Depression is a common condition among long-term care residents with limited treatment options available. There are few nonpharmacological interventions available to this population. This study examined the use of a prescribed, therapeutic recreation-nursing intervention, wheelchair biking, for treatment of symptoms of depression in older adults in a long-term care setting. A classical experimental design was used and was guided by the Roy Adaptation Model. Forty residents were pretested for depression and randomly assigned to two groups. A 2 week trial of biking therapy was provided to the treatment group. All participants were posttested. Findings indicated there was a statistically significant improvement in depression scores for the treatment group and no significant change for the control group. This study contributes to the body of knowledge of nursing regarding options for the treatment of depression in older adults, and is an encouraging indicator that psychosocial interventions may be effective in reducing depression. PMID- 11915270 TI - Development of an intervention to cope with depression following myocardial infarction. A nurse-facilitated cardiac support group. PMID- 11915271 TI - Clinical implications of respiratory system changes in aging. AB - Nurses must consider the many age-related respiratory system changes when assessing and managing respiratory-related symptoms of older individuals. This article reviews upper and lower respiratory tract changes and provides tips for clinical management of the older person. Pertinent respiratory symptom assessment scales are described. Smoking cessation and other health promotion counseling is discussed. PMID- 11915273 TI - Everybody needs a confidant. PMID- 11915272 TI - Patterns of telephone use among nursing home residents. AB - There is convincing evidence that nursing home residents who have more visitors fare better than those who have fewer visitors. However, as many as one third of all individuals living in nursing homes have virtually no visitors. The purpose of this study was to address this concern by examining telephone use in older adults living in nursing homes, and evaluating the potential of telephone communications as a means of social support for this at-risk group. Using a recording device activated each time the receiver was picked up, the research team "listened in" on the telephone conversations of three nursing home residents (aged 76, 79, and 92) for a period of 1 week. The transcripts revealed 56 minutes of actual conversation during the week (10, 21, and 25 minutes, respectively), most often with family or longtime friends living out of town. Each resident laughed aloud more than once per minute. The telephone conversations provided the residents with vivid glimpses of life outside the walls of their facility and appeared to help them stay connected with their family and friends. The findings support further study of the telephone as a way to sustain authentic social support in long-term care populations. PMID- 11915274 TI - Alone in a crowd. A study of social networks in home health and assisted living. AB - This study demonstrated the need for intervention in social network development by both clients of home health and residents of assisted living facilities. With the well-known connection between social networks and health, it is vital that nurses working with these populations be aware of the potential for isolation, screen for it, and, if necessary, develop interventions in the care planning process to address the lack of social networks. PMID- 11915275 TI - How the road to fitness led me to a "fountain of youth". PMID- 11915276 TI - Therapeutic recreation in the nursing home. Reinventing a good thing. AB - The purpose of this article is to increase awareness of therapeutic recreation as a treatment option for nursing home residents. It addresses recent regulations and documentation changes that have occurred under the prospective payment system, and summarizes efficacy research documenting the possible health outcomes of recreation therapy. PMID- 11915277 TI - [Studies on quality and safety control of drugs for human use from transgenic animals/clone animals]. AB - Recently the pharmaceuticals, which were produced using transgenic animals, have been developed, and will be submitted for registration in nearly future in Japan as well as in USA and EU. In addition, clone animals are also thought to be useful for the productions of the pharmaceuticals. This study has been, therefore, undertaken to establish the technical requirement for registration of the pharmaceuticals. They should be evaluated from the following standpoints: 1) Transgene construct and its characterization; 2) Creation and characterization of the transgenic founder animal; 3) Establishment of a reliable and continuous source of transgenic founder animals; 4) Generation and selection of the production animals; 5) Maintenance of transgenic animals; 6) Recovery and purification of products from transgenic animals; 7) Characterization of products; 8) Process validation/evaluation and in-process test; 9) Specification of products; 10) Stability of products; 11) Preclinical safety evaluation and clinical evaluation. Cloning technology by nuclear transfer of a transformed somatic cell has been already applied to the creation of the transgenic founder animal for the production of pharmaceuticals. The pharmaceuticals produced using the clone animals could be evaluated from almost the same standpoints. However, the flexible evaluation will be also needed depending on the development of the technology. PMID- 11915278 TI - [Ergocalciferol Reference Standard (Control 001) of National Institute of Health Sciences]. AB - The raw material of ergocalciferol was examined for the preparation of "Ergocalciferol Reference Standard (Control 001)". Analytical data obtained were: melting point, 114.8 degrees C; UV and infrared spectra, the same as those of JP Ergocalciferol Reference Standard (Control 971); specific absorbance, E1ca1% = 471(265 nm); optical rotation, [alpha]D20 = +102.4 degrees; thin-layer chromatography, no impurities were detected until 100 micrograms; high performance liquid chromatography (HPLC), total amount of impurities estimated to be less than 0.1%. Based on the above results, the raw material was authorized as the Japanese Pharmacopoeia Ergocalciferol Reference Standard (Control 001). PMID- 11915279 TI - [Alprostadil Reference Standard (Control 001) of National Institute of Health Sciences]. AB - The raw material of Alprostadil was examined for the preparation of "Alprostadil Reference Standard (Control 001)". Analytical data obtained were: IR spectrum, same as that of the Alprostadil Reference Standard (Control 923); thin-layer chromatography, no impurities were detected until 20 micrograms; high-performance liquid chromatography (HPLC), total amount of impurities estimated to be less than 0.2%. Based on the above results, the raw material was authorized as the Japanese Pharmacopoeia Alprostadil Reference Standard (Control 001). PMID- 11915280 TI - [Cholecalciferol Reference Standard (Control 001) of National Institute of Health Sciences]. AB - The raw material of cholecalciferol was examined for the preparation of "Cholecalciferol Reference Standard (Control 001)". Analytical data obtained were: melting point, 83.2 degrees C; UV and infrared spectra, the same as those of JP Cholecalciferol Reference Standard (Control 971), respectively; specific absorbance at 265 nm, E1ca1% = 478; optical rotation, [alpha]D20 = +108.6 degrees; thin-layer chromatography, no impurities were detected until 100 micrograms; high-performance liquid chromatography (HPLC), total amount of impurities estimated to be less than 0.05%. Based on the above results, the raw material was authorized as the Japanese Pharmacopoeia Cholecalciferol Reference Standard (Control 001). PMID- 11915281 TI - [The Endotoxin 100 Reference Standard of the National Institute of Health Sciences (the Japanese Pharmacopoeia Endotoxin 100 Reference Standard) (Control 0101)]. AB - To establish the second lot (Control 0101) of the Endotoxin 100 Reference Standard of the National Institute of Health Sciences (the Japanese Pharmacopoeia Endotoxin 100 Reference Standard), a candidate standard (CS) was prepared and then evaluated. The potency of the CS was assayed against USP Endotoxin Reference Standard (Lot G-1) and defined as containing approximately 100 endotoxin units (EU) per vial by a collaborative study in which 5 laboratories participated. Based on the results, the CS was authorized to be the second lot of the Endotoxin 100 Reference Standard containing 100 EU of endotoxin per vial. PMID- 11915283 TI - [Japanese guidelines on research use of human materials and disease information]. AB - Demand on the use of human materials and personal disease information for research and development comes on the stage of action. International competition in biomedical and pharmaceutical research pushed the government and related societies about to release guidelines on the issue to ensure the protection of direct and indirect research participants. To work on human materials and information now requires ethical and scientific review by the research ethical committee and the guidelines set the stage of reviewing process. We should aware of the overall view of situation around and of the guidelines before planning the experiments and analyses on human materials and personal information. In this review we summarized the related guidelines and put comments on them to help researchers to understand the situation. PMID- 11915282 TI - [Hypersensitivity about environmental chemicals--mainly about food allergy]. AB - The hypersensitivity of environmental chemicals and natural products has been reviewed. Among environmental chemicals, small molecular weight molecules work as hapten and cause immediate-type and delayed-type hypersensitivity. Among natural products, relatively lower molecular weight protein or glycoprotein (MW 10,000 70,000 kDa) work as allergen and cause mainly immediate-type hypersensitivity. In recent years, amino acid sequence of important natural allergens have been determined, and three-dimensional structure and IgE epitopes of some of these allergens have also been determined. The characteristics of both inhalation and food allergens have been summarized. As for food allergens, the stability of these proteins in simulated gastric fluid(SGF) was one of the most important characteristics. In the last parts, the approach to the assessment of allergenic potential of genetically modified foods has been summarized. PMID- 11915284 TI - [Teratogenicity study of N-methylphenyl-N'-methylphenyl-p-phenylenediamine (MMPD) in rats by oral administration]. AB - Teratogenicity of N-methylphenyl-N'-methylphenyl-p-phenylenediamine (MMPD), a rubber antioxidant, was examined in Wistar rats. MMPD was given to pregnant rats by gavage once a day from day 6 through day 15 of pregnancy at doses of 0, 2, 4 and 8 mg/kg/day. The pregnant rats were sacrificed on day 20 of pregnancy and their fetuses were examined. MMPD was not maternal toxicity at any dose. MMPD did not affect growth or morphological development of fetuses. MMPD caused early embryonic death at 8 mg/kg. It was concluded from these results that the no observed-adverse-effect level was 2 mg/kg/day for rat fetuses, and 8 mg/kg/day or higher for pregnant rats. The teratogenicity of MMPD was inconclusive due to its embryo-fetal lethality. PMID- 11915285 TI - Isoquinoline alkaloid production by transformed cultures of Papaver somniferum. AB - Three clones of transformed cultures of opium poppy (Papaver somniferum L.) were established by infection with Agrobacterium rhizogenes MAFF 03-01724. MAFF clone 1 being capable of forming somatic embryos was selected and its growth and isoquinoline alkaloid production was investigated. The illumination, temperature and nutrient medium composition greatly affected growth, cell morphology and alkaloid accumulation. The MAFF clone 1 cultured in Root Culture medium in the dark at 22 degrees C accumulated a high quantity of sanguinarine (652 micrograms/g dry weight) though the growth was poor (4.4 fold as fresh weight basis after 2 months of culture). The MAFF clone 1 cultured in a quarter macro salt strength Woody Plant medium under 14 h/day light at 22 degrees C developed into plantlets and accumulated significant quantity of codeine (648 micrograms/g dry wt) together with papaverine, noscapine, and sanguinarine. This clone was applied to a rotating drum fermenter (2 L working volume), and ca. 0.3 mg codeine and 0.06 mg sanguinarine were obtained after 4 weeks of culture. One quarter of the codeine produced was found in the culture medium. PMID- 11915286 TI - [Rapid quantitation of follistatin by surface plasmon resonance (SPR) immunoassay]. AB - A simple, rapid, and accurate assay using surface plasmon resonance (SPR) apparatus with anti-follistatin antibody (SPR immunoassay) has been developed for the quantitation of recombinant follistatin. This assay can be performed with a direct injection of conditioned medium; results were obtained within 10 min. The quantitation component of this assay was precise and accurate with a limit of quantitation of 62.5 ng/ml in Ham's F12 medium containing 2% fetal bovine serum. These results demonstrate that SPR immunoassay is a powerful technique for several researches, especially for screening of gene transfectant and monitoring of protein production. PMID- 11915287 TI - Change in the particle size distribution of poly (L-lactide) wear debris by gamma ray irradiation. AB - It has been known that the wear debris causes failure of implant prostheses. In this study, the convenient wear test of poly(L-lactide) (PLLA) was established and the particle size of PLLA wear debris was analyzed using the Coulter counter. Then, the changes in the particle size distribution of PLLA wear debris by gamma ray irradiation were observed dose-dependently at the dose of 10, 25 and 50 kGy. With the increasing irradiation dose, the particle size distribution of PLLA wear debris shifted toward the smaller diameter size, and the mean diameter of PLLA wear debris significantly decreased. In addition, the tensile strength and the molecular weight of irradiated PLLA were also decreased by increasing the irradiation dose. The lowering of the molecular weight by gamma-irradiation resultingly caused the decreases in tensile strength of irradiated PLLA and the particle size of the wear debris derived from irradiated PLLA. PMID- 11915288 TI - [Study on evaluating methods for the quality control of glycoprotein products. (III)--Erythropoietin products. Part 3]. AB - We reported previously that peptide mapping using high-performance liquid chromatography with electrospray ionization mass spectrometry (LC/MS) and liquid chromatography with tandem mass spectrometry (LC/MS/MS) are useful for determination of the glycosylation sites, carbohydrate structure, and site specific carbohydrate heterogeneity of glycoproteins. Here, with intention to enhance the sensitivity and shorten the time-span of analysis to characterize glycoproteins, especially biotechnological products with carbohydrate moieties, we studied the introduction of HPLC with a microbore column to LC/MS with recombinant erythropoietin (rh-EPO). In addition, we evaluated the ability of LC/MS/MS precursor-ion scanning to make identification of glycopeptides and facilitate the analysis of carbohydrate moieties. We found that the peptide mapping with microbore HPLC is highly sensitive and more rapid than the previous method, and the precursor-ion scanning is helpful for identifying glycopeptides. Our results indicate that these methods are very useful for characterization and quality control of the carbohydrate moieties of biotechnological products. PMID- 11915289 TI - [Estimated production by the official inspection of tar colors (including aluminum lakes) in fiscal year 2000]. AB - There were 176 official inspections of tar colors and their lakes in fiscal year 2000, and 175 samples were qualified. The quantity of tar colors that passed inspection in Japan in fiscal year 2000 reached 137.5 tons. Tar color production is estimated by month and by manufacturer. The food tar color produced in the largest quantity was Food Yellow No. 4, accounting for 43.4% during this period. PMID- 11915291 TI - [Betamethasone Sodium Phosphate Reference Standard (Control 001) of National Institute of Health Sciences]. AB - The raw material of betamethasone sodium phosphate was examined for the preparation of the "Betamethasone Sodium Phosphate Reference Standard (Control 001)". The analytical data obtained were: melting point, 207.2 degrees C; pH, 8.1; optical rotation, [alpha]D20 = +104.4 degrees; UV spectrum, lambda max of 242 nm and specific absorbance in water at 242 nm = 272.9; IR spectrum, specific absorptions at 3386.9, 1721.7, 1663.3, 1620.4, 1605.0, 1094.3, 985.8, 889.8 cm-1; free phosphoric acid, 0.3%; thin-layer chromatography, one impurity was detected until 200 micrograms; high-performance liquid chromatography, total amount of impurities estimated to be less than 0.5%; water, 9.2%. Based on the above results, the raw material was authorized as the Betamethasone Sodium Phosphate Reference Standard (Control 001) of the National Institute of Health Sciences. PMID- 11915290 TI - [Studies on rejected food yellow no. 5 (sunset yellow FCF) aluminum lake]. AB - One out of two sunset yellow FCF aluminum lakes (Y-5Als) did not comply with the specifications in JSFA-VII in the official inspection of tar colors in fiscal year 2000. A sub-spot was detected in the paper chromatography test. This rejected sample was analyzed by HPLC for the subsidiary color, raw materials and intermEdiates in Y-5. The sub-spot was identified as sulfanilic acid azo R salt color, and its content was estimated at 4.5% as the content of Y-5 in Y-5Al being 100.0%. PMID- 11915292 TI - [Hydrocortisone Sodium Phosphate Reference Standard (Control 001) of National Institute of Health Sciences]. AB - The raw material of hydrocortisone sodium phosphate was examined for the preparation of the "Hydrocortisone Sodium Phosphate Reference Standard (Control 001)". The analytical data obtained were: pH, 8.3: optical rotation, [alpha]D20 = +126.2 degrees; UV spectrum, lambda max of 248 nm and specific absorbance in water at 248 nm = 338.6; IR spectrum, same as that of the Hydrocortisone Sodium Phosphate Reference Standard (Control 891); free phosphoric acid, 0.2%; free hydrocortisone, 0.01%; thin-layer chromatography, no impurity was detected until 200 micrograms; high-performance liquid chromatography, total amount of impurities estimated to be less than 0.2%; residual solvent, 0.0% (acetone) and 0.02% (ethanol); loss on drying, 1.5%. Based on the above results, the raw material was authorized as the Hydrocortisone Sodium Phosphate Reference Standard (Control 001) of the National Institute of Health Sciences. PMID- 11915293 TI - [Beclometasone Dipropionate Reference Standard (Control 011) of National Institute of Health Sciences]. AB - The raw material of beclometasone dipropionate was examined for the preparation of the "Beclometasone Dipropionate Reference Standard (Control 011)". The analytical data obtained were: melting point, 208.8 degrees C; optical rotation, [alpha]D20 = +91.7 degrees; IR spectrum, same as that of the Beclometasone Dipropionate Reference Standard (Control 865); thin-layer chromatography, one impurity was detected until 40 micrograms; high-performance liquid chromatography, total amount of impurities estimated to be less than 0.5%; loss on drying, 0.6%. Based on the above results, the raw material was authorized as the Beclometasone Dipropionate Reference Standard (Control 011) of the National Institute of Health Sciences. PMID- 11915294 TI - [Dexamethasone Sodium Phosphate Reference Standard (Control 001) of National Institute of Health Sciences]. AB - The raw material for dexamethasone sodium phosphate was examined for the preparation of the "Dexamethasone Sodium Phosphate Reference Standard (Control 001)". The analytical data obtained were: pH, 8.0; optical rotation, [alpha]D20 = +79.6 degrees; UV spectrum, lambda max of 242 nm and specific absorbance in water at 242 nm = 313.6; IR spectrum, same as that of the Dexamethasone Sodium Phosphate Reference Standard (Control 893); free phosphoric acid, 0.06%; free dexamethasone, 0.07%; thin-layer chromatography, no impurities were detected until 100 micrograms; high-performance liquid chromatography, total amount of impurities estimated to be less than 0.2%; residual solvent, 4.3% (ethanol); water, 7.3%. Based on the above results, the raw material was authorized as the Dexamethasone Sodium Phosphate Reference Standard (Control 001) of the National Institute of Health Sciences. PMID- 11915295 TI - [Glycyrrhizinic Acid Reference Standard (Control 001) of National Institute of Health Sciences]. AB - The raw material of glycyrrhizinic acid was examined for preparation of the "Glycyrrhizinic Acid Reference Standard". The analytical data obtained were: UV spectrum: lambda max, 251 nm; and specific absorbance (E1ca1%) in ethanol at 251 nm, 146; IR spectrum, specific absorptions at 1714, 1655, 1215, and 1170 cm-1; and the spectrum of raw material was consistent with that of Standard (Control 991). Also, thin-layer chromatography, no impurity was detected; high-performance liquid chromatography, several impurities were detected. The amount of each impurity was estimated at less than 0.2% and total amount of impurities was less than 0.4%. Based on the above results, the candidate material was authorized as the Glycyrrhizinic Acid Reference Standard (Control 001) of the National Institute of Health Sciences. PMID- 11915296 TI - [Berberine Hydrochloride Reference Standard (Control 001) of National Institute of Health Sciences]. AB - The raw material of Berberine Hydrochloride was examined for preparation of the "Berberine Hydrochloride Reference Standard". The analytical data obtained were: UV spectrum: lambda max, 420, 345, 263 and 228 nm and specific absorbance (E1ca1%) in ethanol at each lambda max, 155, 724, 796 and 820, respectively; IR spectrum, specific absorptions at 2844, 1635, 1569, and 1506 cm-1; and the spectrum of raw material was consistent with that of Standard (Control 941). Also, thin-layer chromatography, an impurity was detected; high-performance liquid chromatography, several impurities were detected. The amount of each impurity was estimated at less than 0.1% and the total amount of impurities was less than 0.2%. Based on the above results, the candidate material was authorized as the Berberine Hydrochloride Reference Standard (Control 001) of the National Institute of Health Sciences. PMID- 11915297 TI - Measurement of pulpal blood flow in dogs with nonradioactive colored microspheres. AB - The viability of dental pulp depends largely on regional blood flow, as in other organs. Measurement of absolute pulpal blood flow (PBF) and comparisons with blood flow in other organs allow the prediction of microvascular regulation in dental pulp. In previous studies, PBF was measured in dogs mainly with radioactive microspheres. However, this established technique is inaccessible to many investigators due to concerns over radiation safety and radioactive waste. To overcome these limitations, a new method has been introduced that involves the use of nonradioactive colored microspheres for measuring regional blood flow in the myocardium and in other organs in animals. However, no previous studies have investigated the use of this method to measure PBF in dogs. We attempted to determine whether blood flow in dental pulp, which comprises a small amount of the total tissue in dogs, could be measured using this technique by comparing the measured values with those for regional myocardial blood flow. Mean blood flow values were between 0.148 and 0.182 ml/min/g for dental pulp at four different sites and about 1.0 ml/min/g in regional myocardium. These values are comparable to those previously reported using radioactive microspheres. As nonradioactive colored microspheres safety permitted measurement of absolute PBF in dogs, this technique appears to be useful for research into microvascular blood flow in dental pulp. PMID- 11915298 TI - Allelic imbalance on the long arm of chromosome 21 in human oral squamous cell carcinoma: relationship between allelic imbalances (LOH and MSI) and clinicopathologic features. AB - Frequent allelic imbalances, including loss of heterozygosity (LOH) and microsatellite instability (MSI), have been found on the long arm of chromosome 21 (21q) in several types of human cancer. This study was designed to identify the tumor suppressor locus (or loci) associated with oral squamous cell carcinoma (SCC) on 21q. In order to understand the details of genetic alterations on chromosome 21, we performed polymerase chain reaction analysis of microsatellite polymorphisms corresponding to ten loci on this chromosome. We examined forty primary tumor tissues, forty corresponding normal tissues, and seven lymph node metastatic tissues. We identified novel tumor suppressor loci in this region in primary oral SCCs. To further determine the role of 21q deletions in oral cavity carcinogenesis, forty oral SCCs were examined for allelic imbalances (LOH or MSI) at 21q using ten microsatellite markers. Among these forty patients, twenty-six (65%) showed LOH at one or more loci. Deletion mapping of these tumors revealed four discrete, commonly deleted regions on the chromosome arm. Furthermore, we detected MSI in seventeen of those tested cases (42.5%). We compared our results with the clinicopathologic features. A number of sites displaying LOH at 21q could be detected in early stage lesions, and the frequencies of LOH tended to be higher in later clinical stages, but no statistical correlation was observed. Our results strongly suggest that allelic imbalances on 21q are involved in the development of oral SCC and that at least four different putative tumor suppressor genes contributing to the pathogenesis of this disease are present on 21q. Furthermore, allelic loss on 21q appears to be a useful indicator for evaluating the malignancy and prognosis of oral SCC, because the LOH of recurrent cases was more frequent than that of non-recurrent ones. PMID- 11915299 TI - Characteristics of newly formed bone during guided bone regeneration: observations by immunohistochemistry and confocal laser scanning microscopy. AB - The purpose of this study was to investigate the characteristics of new bone formation during guided bone regeneration (GBR) using immunohistochemistry and confocal laser scanning microscopy. e-PTFE membranes were applied to defects created in the tibiae of rats, and some animals were sacrificed 6, 8, or 10 days later. Serial paraffin sections were cut, stained with H-E, and examined to analyze the ratio of new bone formation. Immunohistochemical staining with a monoclonal antibody specific for PCNA was used to evaluate the proliferating activity. In other experimental rats, calcein was injected at 6, 8, and 10 days after the surgery, and the animals were sacrificed 48 hr after injection. Their tibiae were removed, and Villanueva bone staining was performed before observation using confocal laser scanning microscopy to investigate the mineralization of new bones. The bone occupation ratio increased day by day, but the experimental groups had significantly higher ratios than control groups (without membrane) at each of the time periods. However, PCNA positive cells decreased over time in all groups, and there were no significant differences among the groups. Mineralization occurred more rapidly in the experimental groups than in the control groups. These results suggest that GBR accelerates the migration of osteogenic cells, the formation of new bone, and mineralization in the defect created by the e-PTFE membrane. PMID- 11915300 TI - Decay in prepulse facilitation of calcium channel currents by Gi/o-protein attenuation in hamster submandibular ganglion neurons, but not Gq/11. AB - The calcium ion influx through voltage-dependent calcium channels (VDCCs) has a vital role in the control of neurotransmitter release and membrane excitability. Prepulse facilitation is a phenomenon in which a strong depolarizing pulse induces a form of the VDCCs that exhibits an increased opening probability in response to a given test potential; this persists for several seconds after repolarization. It has been reported that prepulse facilitation occurs via dissociation of the guanosine triphosphate (GTP)-binding proteins (G-proteins) from the VDCCs and that recovery from facilitation involves rebinding of the G proteins. The heterotrimeric G-proteins act as switches that regulate information processing circuits connecting cell surface G-protein-coupled-receptors to a variety of effectors. In this study, we have studied the characterization of G protein subtypes in prepulse facilitation of VDCCs currents (Ica) in hamster submandibular ganglion (SMG) neurons, using whole-cell patch clamp recordings. Under control conditions, with GTP (0.1 mM) in the recording pipette, the rate of prepulse facilitation was 19.0 +/- 1.9% (n = 13). Intracellular dialysis with GDP beta-S (0.1 mM), G-protein blocker, and pretreatment of neurons with N ethylmaleimide (NEM) (100 microM for 2 min), Gi/o blocker, attenuated the rate of prepulse facilitation. Intracellular dialysis of anti-Gq/11-antibody did not alter it. These results suggest that prepulse facilitation of VDCCs is due to Gi/o-types of G-protein, but not to the Gq/11-type, in SMG neurons. PMID- 11915301 TI - Comparison of the activity of four chitosan derivatives in reducing initial adherence of oral bacteria onto tooth surfaces. AB - We examined the effects of four kinds of chitosan derivatives on initial adherence of oral bacteria onto human anterior teeth surfaces. The buccal surfaces of anterior teeth were used as the experimental surfaces. They were divided into five rectangle areas with outer dimensions of about 2 mm x 4 mm. After applying two ml of a sample solution onto the tooth surfaces, an examiner wiped each rectangle area with a sterilized plastic swab one, three and six hours later. Then we measured bacterial counts in sterilized swabs with mitis salivarius agar. We found that the order of magnitude of the inhibitory effect on the adherence of oral bacteria was low molecular chitosan > phosphorylated chitosan > amorphous chitosan > carboxymethyl chitosan. The solution containing 0.5% low molecular chitosan depressed the bacterial adherence to the same extent as a 50 ppm chlorhexidine digluconate solution for three hours, and 0.1% phosphorylated chitosan also exhibited an inhibitory effect in bacterial adherence for one hour. Amorphous chitosan had a moderate inhibitory effect, but no clear inhibitory activity was found with 0.1% carboxymethyl chitosan. These results suggest that low molecular chitosan and phosphorylated chitosan have the potential to effectively inhibit the initial adherence of oral bacteria onto human tooth surfaces. PMID- 11915302 TI - Effect of rinsing with phosphorylated chitosan on four-day plaque regrowth. AB - This clinical investigation examined the effect of phosphorylated chitosan rinsing on plaque development and on the buffering capacity of plaque suspension. Three male adult subjects participated in the trial that was designed as a single blind study. Participants refrained from mechanical oral hygiene procedures during a four-day study and rinsed three times a day with 20 ml of test solutions. A wash-out period of three days was instituted between the placebo and phosphorylated chitosan rinsing period. Clinical evaluation and plaque sampling were performed at the end of each test period. We disclosed plaque accumulations on the buccal upper front teeth with a two-tone disclosing agent to distinguish between newly formed plaque and old plaque. After taking color slides, we then used a computerized image analysis. Tooth areas covered by plaque on the color slides were digitized and expressed as percentages of the tooth area. The buffering capacity of the collected plaque fluid was determined by using a beta titrator. A mouth rinse containing 0.5% phosphorylated chitosan significantly reduced both newly formed plaque areas (red disclosed; p < 0.001) and old plaque areas (blue disclosed; p < 0.01) compared to a placebo rinsing. However there was no significant difference in the plaque buffering capacity (p > 0.05) between the mouth rinse containing 0.5% phosphorylated chitosan and placebo. These findings might suggest that mouth rinse containing phosphorylated chitosan would be effective in reducing plaque formation and have a slight ability to enhance plaque buffering capacity. PMID- 11915303 TI - [Alteration of neuronal activities following repeated administration of stimulants]. AB - It has been well-known that abuse of psychostimulants such as amphetamine and methamphetamine (MAP) induces behavioral sensitization (reverse tolerance) to MAP, resulting in psychotic effects such as hallucinatory-delusional state. Animals treated with MAP repeatedly also show the behavioral sensitization to MAP. This paper focuses on the pathogenesis and mechanism underlying sensitization to MAP after repeated treatment with MAP. MAP is known to release dopamine (DA), noradrenalin (NA) and serotonin (5-HT), and bind with the same sites on DA-, NA- and 5-HT-transporters as do these monoamines, thereby inhibiting re-uptake of these substances. As a result, these monoamines accumulate in the synaptic areas unnerved by the monoamine systems. An increase in the monoamines also occurs in the dendritic areas of DA, NA and 5-HT cells probably by a mechanism similar to those in the presynaptic terminals of monoamines. Releases and syntheses of DA, NA and 5-HT are inhibited by the monoamine per se via their autoreceptors such as D2, alpha 2 and 5-HT1A receptors, respectively. It is noteworthy that repeated MAP treatment results in the reduction of DA transporters, and such a decrease in transporters has been also found in MAP abusers by PET studies, suggesting a decrease in DA transporters is related with the appearance of reverse tolerance. Repeated MAP administration induces immediate early gene such as c-fos, c-jun and arc, and the increase in arc is inhibited by D1 and NMDA antagonists, suggesting an important role of such genes in inducing reverse tolerance. In electrophysiological studies using anesthetized rats treated with MAP repeatedly, hyposensitivities and hypersensitivities to DA and MAP have been found in nucleus accumbens receiving dopaminergic input from ventral tegmental area, 24-30 h and 5 days after the final administration of MAP, respectively, although the sensitivities recovered to the normal level 10 days after the treatment. The hypersensitivities were probably mediated via D1/D2 receptors. Thus, the hypersensitivities of nucleus accumbens neurons to DA and MAP are actually completed after repeated treatment of MAP. Therefore, it is of great interest to elucidate the molecular mechanism responsible for the DA receptor hypersensitivity. PMID- 11915304 TI - Prevalence of solvent inhalation among junior high school students in Japan and their background lifestyle: results of Chiba prefecture survey 1994. AB - To estimate the lifetime prevalence of solvent abuse among general junior high school students in Chiba prefecture and to understand their background lifestyle, in 1994 the author surveyed 6,795 students enrolled in 15 junior high schools. The lifetime prevalence of solvent abuse was 1.9% in males, 1.1% in females and 1.5% of the total. This implied that the lifetime prevalence of solvent abuse was decreasing year-on-year in males, had recovered to the 1990 level in total, but had stagnated between the 1990 level and the 1992 level in females. The regularity of daily life was more significantly disturbed in Lifetime Users than in Non-Users. School life and family life were significantly less relaxed for Lifetime Users than for Non-Users. Cigarette smoking and some types of alcohol drinking were strongly related to solvent abuse. More Lifetime Users than Non Users were aware of harmful effects of solvent abuse. The author suggests that developing practical educative measures to promote behavioral change, as well as increasing the frequency with which the entire family eats dinner together, is necessary. PMID- 11915305 TI - [Drinking behavior of female nursing students and stress factors related problem drinking]. AB - The survey was done to clarify how the behavior of drinking alcoholic beverages of female nursing students relates to the stress they feel in their school life. The questionnaires were sent to 337 female students of two nursing schools located in metropolitan area. Students were asked to reply the questions of Adolescent Alcohol Involvement Scale (AAIS), and other questions related to their smoking habits and stress felt in their school life. Alcohol misusers who scored 42 points or more on AAIS were 13.7% of the valid respondents and non-drinkers and those who scored 19 and under on the scale were 4.1%. Those who drink more than over per week accounted for 12.0%, and those who have more than 3 drinks at one time accounted for 61.5%. Those who experienced alcohol drinking during childhood replied that this was encouraged by their parents. Those parents were found to be very generous about children's drinking at home later. The analysis of stress factors in students' life in relation to drinking behavior revealed that the students with scores on AAIS above 42 tended to show less eagerness in studying, and that clinical practice and report writing did not give than much stress. PMID- 11915306 TI - [Clinical study of BRON-L syrup (cough suppressant) abuse]. AB - In 1980s, abuse and dependence of BRON-W syrup (cough suppressant), which contains methylephedrine, dihydrocodeine, chlorpheniramine and caffeine, were prevalent in Japan. Pharmacological and clinical studies suggest that methylephedrine and dihydrocodeine cause dependence. Although BRON-L syrup, newly modified cough suppressant contains only chlorpheniramine and caffeine, there still are abuse and dependence of this drug. In this report, three cases of BRON L syrup abuse are demonstrated. All cases started using BRON-L syrup in the late teens in their peer groups, and dropped out from school. Case 1 misused only BRON L syrup, but case 2 and 3 were multi-drug abusers (case 2: amphetamine, cocaine, and marijuana, case 3: solvent, alcohol, bromovalerylurea), and had kept in tough with the peer groups. Case 2 and 3 hospitalized more than 2 times. Withdrawal symptoms, such as headache, insomnia, and irritability were mild and improved in a few weeks after drug use was stopped. These findings suggest that 1) psychosocial backgrounds of these cases are in common with those of BRON-W syrup abusers, but 2) the clinical course and prognosis of multi-drug abusers are different from the BRON single abuser, 3) chlorpheniramine and caffeine possibly cause dependence, 4) abusers are likely to choose BRON brand although two main dependence-producing constituents are removed from it now. Therefore, prevention and care of BRON-L abusers requires both psychosocial and pharmacological aspects. PMID- 11915307 TI - [Significance of 99mTc-labeled perfusion agents in the simultaneous assessment of myocardial perfusion and cardiac function]. AB - Simultaneous assessment of left ventricular myocardial perfusion and systolic function was accomplished by utilizing ECG-gated myocardial perfusion SPECT. This development in nuclear cardiology will be attributed to the recent advances in new 99mTc-labeled perfusion agents, multi-detector SPECT system and software for automatic edge-detection of the left ventricle. In this article, we described about the clinical utilities of this method in detecting "hibernating myocardium," severe coronary artery disease patients with exercise-induced LV dysfunction, in predicting functional recovery after reperfusion therapy for acute myocardial infarction patients and in diagnosing patients with right heart diseases. PMID- 11915308 TI - [Estimation and optimization of the use of standard arterial input function for split-dose administration of N-isopropyl-p[123I]iodoamphetamine]. AB - Use of a standard arterial input function and calibrating it by a single blood sample or a continuous arterial blood sample has been researched for a repeat CBF assessment with split-dose administration of N-isopropyl-p[123I]iodoamphetamine (IMP). METHODS: The study population consisted of 5 normal volunteers and 5 patients with cerebrovascular disease. IMP was injected twice (111 MBq/2 ml each) into the anti-cubital vein at a constant infusion speed for 1 min. The arterial input function was monitored during the study including a continuous measurement of radioactivity concentration of both the whole-blood and the octanol-soluble component (Real-Input Function, RIF). Standard input function was determined, and was calibrated either by a single blood sample or a continuous blood sample to estimate the Estimated-Input Function (EIF). Area-Under-the Curve (AUC) was then compared between RIF and EIF. RESULTS: In case EIF was estimated with a single blood sample, the minimum error of estimated AUC was obtained when calibrated at 7 minutes after either the 1st or 2nd injections. Deviation of AUC for [0, 30] was +/- 6.6%, and +/- 5.0%, respectively. If calibrated with a continuous blood sample, the minimum error of AUC with the continuous blood sampling period of 10 min for [0, 30] and [30, 60] was +/- 5.3% and +/- 4.0%, respectively. CONCLUSIONS: AUC of EIF with either a single or continuous blood sampling appeared to have reasonably small errors, suggesting the validity of the use of standardized input function in the split-dose IMP SPECT. PMID- 11915309 TI - [Assessment of left ventricular function by thallium-201 quantitative gated cardiac SPECT]. AB - PURPOSE: Present study was designed to evaluate the accuracy of the measurement of left ventricular volume by quantitative gated SPECT (QGS) software using 201T1 and the effect of cutoff frequency of Butterworth prereconstruction filter on the calculation of volume. METHODS: The RH-2 type cardiac phantom and 20 patients with ischemic heart disease were studied. Left ventricular end-diastolic volume (EDV), end-systolic volume (ESV) and ejection fraction (EF) were calculated by the QGS software using the various frequency of Butterworth filter. These parameters were evaluated by Simpson's method using left ventriculography (LVG). RESULTS: The volume of the phantom calculated by QGS was under-estimated by 14%. In the clinical study, EDV and ESV measured by QGS were smaller than those obtained from LVG by 10%. When the cutoff frequency of Butterworth filter was 0.43 cycles/cm, the values measured by QGS were best correlated with those by LVG (EDV: r = 0.80, p < 0.001; ESV: r = 0.86, p < 0.001; EF: r = 0.80, p < 0.001). CONCLUSION: These data suggest that 201Tl quantitative gated cardiac SPECT can estimate myocardial ischemia and left ventricular function simultaneously. PMID- 11915310 TI - [No 123I-BMIPP accumulation in the myocardium and type I CD36 deficiency in a patient with acute subendocardial infarction: a case report]. AB - This patient was a 70-year-old man had acute subendocardial infarction in the inferior wall. 123I-BMIPP myocardial scintigraphy showed no accumulation in the myocardium. 123I-MIBG myocardial scintigraphy on the early and delay images and 99mTc-tetrofosmin myocardial scintigraphy at rest showed slightly decreased accumulation of the tracer in the apical region and in middle inferior wall of the left ventricle, indicating subendocardial infracted area. In the examinations of CD36 in platelets and monocytes, the patient had negative CD36 in platelets and monocytes, and type I CD36 deficiency was diagnosed. We supposed that no 123I BMIPP accumulation may be related closely to type I CD36 deficiency. PMID- 11915311 TI - [Multi-center study for the evaluation of clinical usefulness of attenuation and scatter correction on 201Tl myocardial SPECT]. AB - The aim of this study was to evaluate the clinical usefulness of attenuation and scatter correction (AC, SC) on a 201Tl myocardial single-photon emission computed tomography (201Tl SPECT) as a multi-center trial. With a dual-detecter and a triple-detector SPECT systems with a 99mTc transmission source, simultaneous transmission/emission tomography (TCT/ECT) was performed on 38 patients with angiographically coronary heart disease (CHD) and 26 patients without evidence of CHD. Stress and delayed attenuation and scatter corrected images (SAC) and uncorrected images (NC) were reconstructed. On NC images of normal cases, influence of attenuation was greater in male than female. In comparison of 201Tl distribution between male and female, significant decrease in 201Tl activity was observed in the inferoposterior wall in male and that was observed in the anterobasal wall of the left myocardium in female. Such a difference in 201Tl distribution between male and female disappeared on SAC images. On the diagnostic performance for the identification of CHD, SAC images demonstrated improved specificity and accuracy values in the right coronary arterial territory (RCA) with visual analysis statistically. Sensitivity value in the RCA was also improved, but it was not statistically significant. Sensitivity value in the left circumflex arterial territory (LCX) increased without decrease in specificity value on SAC images. In the left anterior descending arterial territory (LAD), sensitivity value increased on SAC images. Although specificity value decreased on SAC images in LAD territory, it was not statistically significant. The difference in 201Tl distribution between male and female is improved in normal cases by attenuation and scatter correction on 201Tl myocardial SPECT. Diagnostic performance of CHD is also improved by attenuation and scatter correction, especially in territories of which specificity in assessing the absence of disease have been suboptimal. In conclusion, attenuation and scatter correction on 201Tl myocardial SPECT is considered to be clinically useful. PMID- 11915312 TI - [Performance evaluation of dual-head coincidence gamma camera for positron imaging]. AB - AIM AND METHODS: We assessed the performance evaluation of a dual-head coincidence gamma camera (Millennium VG, General Electric, Milwaukee, USA) as a positron emission tomography (PET) scanner based on the measurement indicator for performance evaluation of PET device. RESULTS: The axial spatial resolution was 4.98 mm FWHM at the center of the field of view. The true high count-rate characteristic curve was relatively straight within the range 0-0.2 microCi/ml. The relative recovery coefficient was 0.1 with a diameter of 10 mm and 0.39 with a diameter of 20 mm. CONCLUSION: In this study, we had to make a some change in the protocols described in the measurement indicator for performance evaluation of PET devices. PMID- 11915313 TI - [Nuclear gastroenterology]. AB - Application of nuclear medicine for gastroenterology, especially the liver, portal circulation and alimentary tract was described. In the liver, radiocolloid scintigraphy is useful for the diagnosis and follow-up study of chronic liver diseases. 99mTc-GSA scintigraphy plays a role for the evaluation of hepatic functional reserve. 99mTc-PMT used for hepatobiliary imaging, is also useful for the diagnosis of hepatic tumor and extra-hepatic metastasis of hepatocellular carcinoma. In the study of portal circulation, various administration sites such as intrasplenic, rectal, oral, and intravenous have been reported. In the evaluation of motility function of alimentary tract, estimation of gastric emptying time is well known. Condensed image created from serial esophageal scintigrams using computer processing is also useful for the quantitative and qualitative evaluation of esophageal motility function and gastro-esophageal reflux. Abdominal scintigraphy with 99mTc-human serum albumin enables imaging diagnosis of protein-loosing gastroenteropathy. PMID- 11915314 TI - [Comparison of serum concentrations of trace elements in healthy free-living and hospitalized oldest-old]. AB - PURPOSE: The aim of this study was to examine differences in concentration of trace elements in serum between healthy free-living and hospitalized oldest-old people aged > or = 85 years. SUBJECTS AND METHODS: Subjects were 33 healthy free living oldest-olds (18 men, 15 women, aged 85-91 years, mean age: 87.3 +/- 1.7 years) selected at annual health examination and 40 hospitalized oldest-olds (14 men, 26 women, aged 85-92 years, mean age: 87.8 +/- 2.2 years). The serum concentrations of managanese (Mn), zinc (Zn), copper (Cu), iron (Fe), calcium (Ca), magnesium (Mg) and phosphorus (P) (microgram/ml) were measured using atomic absorption spectrophotometry and colorimetric method. Differences between groups for serum concentration of trace elements were assessed with the Wilcoxon rank sum test. Spearman's rank correlation coefficients were calculated with reference to clinical examination data in the two groups. RESULTS: 1) The concentrations of Ca, Mg, P and Zn were significantly higher in the healthy free-living than in the hospitalized, especially in the P and Zn case. The Cu/Zn ratio was significantly lower in the healthy free-living. 2) The Zn concentration was positively correlated with the Ca concentration, and the P concentration with the total cholesterol level in both groups. Zn was one of the elements demonstrating good correlations with other parameter. Especially in the healthy free-living, the Zn concentration was high positively associated with the Fe concentration. CONCLUSION: These results suggest that the serum concentrations of trace elements in the healthy free-living are higher than in the hospitalized oldest-old, especially in the P and Zn case. PMID- 11915315 TI - [Focus group investigation on expectations of elderly people from health care facilities and physicians]. AB - INTRODUCTION: While the elderly are intensive health care users, little is known about their expectations of various medical facilities and physicians. We investigated their views on advAntages and disadvantages of different medical settings. METHODS: We established four focus groups and surveyed elderly people living in a rural area accessible to local practitioner's offices, and community, regional, and university hospitals, The focus groups examined advantages and disadvantages of care from these facilities. A coding scheme was used to organize the data in thematic categories, In a mail survey, participants ranked the importance of specialty care; overall care; triage of patients; gentle personality; listening skills; and explanation skills, The data were analyzed for statistical differences between the various settings. RESULTS: Total of 41 elderly participated in focus groups, and 40 individuals (response rate 98%) returned statistical surveys. Advantages of care from a primary care doctor included continuity and physician accommodation of patients' preferences, while limited medical services and urgent access were disadvantages. Advantages of the community hospital included expanded medical facilities and having doctors easy to talk with, while disadvantages included lack of trust and frequent turnover. Advantages of larger facilities included extensive medical equipment, a broad range of departments and specialists, and more expert care. The survey revealed expectations for expert triage skills and familiarity with a doctor who is easy to talk with, particularly primary care doctor. DISCUSSION: These elderly subjects expect primary care and community doctors to be accessible and to exhibit excellent behavioral and communication skills, They expect more specialized and technical care in larger hospitals. PMID- 11915316 TI - [On the exacerbation of the systems of fetal Minamata disease patients an urgent proposal of the urgent follow up study for the victims by environmental health hazards]. PMID- 11915317 TI - [Appropriate outcome measures for evaluating change in activities of daily living of elderly residents]. AB - PURPOSE: To identify functional dimensions related to changes in activities of daily living (ADL) and appropriate outcome measures for evaluating change in ADL of residents in a health care home for the elderly. SUBJECTS AND METHODS: The subjects were 123 residents living for three months or more in a health care home for the elderly in Osaka Prefecture. ADL on admission and three months these after were evaluated using four standardized outcome measures: ADL criteria for disabled elderly people developed by the Ministry of Health and Welfare (ADL criteria), the Barthel Index (BI), the Functional Independence Measure (FIM), and the Office of Population Censuses and Surveys (OPCS) scale. ResponsivenesS was also determined with the same four measures. The subjects were divided into two groups according to the frequency of functional training: twice or more and once or less per week. Appropriateness of the outcome measures was determined by examining score distributions and floor and ceiling effects, and by correlating changes in scores with frequency of functional training. RESULTS: The floor effect of the FIM was the smallest among the four outcome measures. The average FIM score on admission was significantly higher than that three months these after (70.7 +/- 30.8 and 71.6 +/- 30.6, respectively P < .05). In terms of funcTional dimensions of the BI, FIM and OPCS measures, the average FIM scores for transfer, walking, and stairs on admission were significantly higher than three months after admission (4.6 +/- 1.8 and 4.8 +/- 1.7 for transfer, 3.8 +/- 2.3 and 4.0 +/- 2.2 for walking, 3.0 +/- 2.1 and 3.2 +/- 2.1 for stairs, respectively, P < .05). Total FIM scores for the same three dimensions were the most responsive among the four outcome measures. Furthermore, when improvement in ADL was evaluated in terms of changes in total scores for the three FIM functional dimensions, those who received functional training twice or more per week showed a significantly higher rate of improvement in ADL than those who received functional training less frequently. CONCLUSION: FIM was found to be the most appropriate ADL measurement for evaluating changes in ADL of elderly residents. Regarding functional dimensions, locomotion in terms of transfer, walking and stairs was the most responsive. These findings indicate that the FIM functional dimension of locomotion is the most suitable for evaluating changes in ADL of elderly residents. PMID- 11915318 TI - [Utility of the public health approach for the world population strategy]. AB - OBJECTIVE: The populations of developing countries have so dramatically increased over the past two centuries that it has tended to undermine sustained economic growth and development. The population growth rate in the Philippines, in particular, remains at 2.0%, one of the highest in the other southeastern Asian countries. This has exacerbated problems of economic disparity and deterioration in development so that the Philippine Government has declared it a political priority to address the population problem and for this purpose has asked the Japan International Cooperation Agency (JICA) for assistance. METHOD: Based on past experience with the world population issue, the International Conference on Population and Development in Cairo (ICPD) declared that the population strategy should be merged into socio-economic development strategies that aim to achieve basic human needs, such as, education, public health and gender equality. JICA assistance to improve maternal and child health status and promote community health initiatives in the Philippines is based on this new paradigm; the ICPD plan. OUTPUT AND CONCLUSION: JICA has supported a diverse range of public health activities, such as the establishment of a comprehensive maternal and child health care system, advocacy of reproductive health, performance of adolescence health education and assistance for participatory community action. The efforts were generally geared towards sustainable implementation of health activities in the community. The JICA project employed the public health approach that gained a global consensus at Cairo to challenge the problem of a continuously high rate of population growth. PMID- 11915319 TI - [Well-being and depression in residents of a provincial city 65 years old or older]. AB - OBJECTIVE: The objective of this study was to examine the relationship between feeling of well-being and clinical depression for a large number of residents in a provincial city in northern Tohoku. The research sought to clarify whether one's lifestyle might play a role in whether one suffered a state of clinical depression. METHODS: The study used survey methods to question every resident 65 years old or older living in the provincial city. The questionnaire was of self registering type and was sent out from October to December, 1998. The content of the survey followed the determination of independence in Activities of Daily Living (ADL), feeling of well-being, the Zung's Self-rating Depression Scale (SDS), and similar items. RESULTS: The results of the study indicated that the morale score (Feeling of well-being) for men was significantly higher at all ages when compared to scores for women, while the Zung index for men tended to be lower than that for women. Although the Zung index tended to rise with age in both sexes, only in women did the morale score tend to decine with increasing age. Comparing men with women in terms of the partial correlation coefficient between the morale score and Zung index, that for men was -0.5856 (P < 0.001) and for women was -0.6363 (P < 0.001). CONCLUSION: It is apparent that a state of depression is not normally suffered if an individual has a feeling of well-being. Moreover, important factors recognized for healthy aging include the general health status, activity level, and the economic situation. Various measures that must be taken to that end, involve fashIoning a way of life for senior citizens so that they form healthy habits, participate in social activities, and take part in sports. PMID- 11915320 TI - [Microdomain: history and perspective]. PMID- 11915321 TI - [Lipid raft domains in sphingolipid-deficient mutants]. PMID- 11915322 TI - [Microdomains and caveolin]. PMID- 11915323 TI - [Neural cell membrane rafts and signal transduction]. PMID- 11915324 TI - [Neural cell adhesion molecules in lipid microdomains]. PMID- 11915325 TI - [Pathological implications of GM1 ganglioside-bound amyloid beta-protein in Alzheimer's disease]. PMID- 11915326 TI - [Interactions between amyloid beta--protein and gangliosides]. PMID- 11915327 TI - [Cellular functions and signal transduction through glycosignaling domain: lactosylceramide mediated superoxide generation from neutrophils]. PMID- 11915328 TI - [GEM (glycosphingolipid enriched microdomain) and cancer]. PMID- 11915329 TI - [Biological significance of lactosylceramide branching and domain formation: glycosphingolipids govern gene expression]. PMID- 11915330 TI - [Association of Fc gamma receptor with low-density detergent-resistant membranes is important for crosslinking-dependent initiation of the tyrosine phosphorylation pathway and superoxide generation]. PMID- 11915331 TI - [T cell membrane raft: its structure and function]. PMID- 11915332 TI - [Biological function of sphingolipids involved in cytokinesis]. PMID- 11915333 TI - [Sphingolipid biosynthesis and membrane traffic]. PMID- 11915334 TI - [Biosynthesis of sphingolipids]. PMID- 11915336 TI - [Conditional knockout mouse of sphingolipid biosynthesis]. PMID- 11915335 TI - [Glia-derived L-Ser regulating survival development and lipid synthesis of developing central neurons]. PMID- 11915337 TI - [Cultured cell mutants defective in synthesis of membrane lipids: a key tool for studies on membrane lipid domains]. PMID- 11915338 TI - [Sphingosine 1-phosphate in yeast]. PMID- 11915339 TI - [Signal transduction of ceramide]. PMID- 11915340 TI - [Crosstalk between ceramide and phosphatidylinositol-3 kinase as a mechanism for cell death/survival determination]. PMID- 11915341 TI - [Involvement of p53 in ceramide signaling cascade]. PMID- 11915342 TI - [Molecular evolution of neutral ceramidase: signalling molecule and virulence factor]. PMID- 11915343 TI - [Localization of sphingomyelinase to Triton X-100-insoluble microdomain and its functional importance]. PMID- 11915344 TI - [Role of glucosylceramide synthase as negative regulator for ceramide]. PMID- 11915345 TI - [Current studies on a novel lipid mediator, sphingosine 1-phosphate, and its receptors]. PMID- 11915346 TI - [Distribution of sphingosine 1-phosphate in plasma lipoproteins and its role in the regulation of the vascular cell functions]. PMID- 11915347 TI - [Sphingosine 1-phosphate and vascular endothelial cells]. PMID- 11915348 TI - [Regulation of Rho family G proteins and cell motility by the Edg family of sphingosin 1-phosphate receptors]. PMID- 11915349 TI - [N-glycosylation of sphingosine 1-phosphate receptor, Edg-1, and its role on receptor internalization through membrane microdomain]. PMID- 11915350 TI - [Molecular diversity of sphingosine kinase]. PMID- 11915351 TI - [Sphingolipid probe]. PMID- 11915352 TI - [Proteins which recognize sphingomyelin]. PMID- 11915353 TI - [Chemical syntheses of sphingophospholipids and their analogues]. PMID- 11915354 TI - [Phage library]. PMID- 11915355 TI - [Construction of the functional glycolipid-cluster]. PMID- 11915356 TI - Effect of low calcium diet on the ultrastructure of the rat parathyroid gland. AB - Young female rats were fed with normal (1.18%) or low (0.05%) calcium diet for 3, 7, 15 or 30 days. The morphology of the parathyroid glands was studied together with serum calcium, parathyroid hormone (PTH), calcitonin and bone mineral density (BMD). As compared to the animals fed with the normal calcium diet, BMD of whole body of the rats fed with the low calcium diet was significantly decreased, whereas the serum PTH level was increased. The parathyroid glands in the rats fed with the low calcium diet were markedly enlarged. In the parathyroid chief cells of the rats fed with the low calcium diet, the Golgi complexes and the cisternae of the granular endoplasmic reticulum were well developed, while the large granules and large vacuolar bodies decreased. Some secretory granules located near the plasma membrane. A proportionally larger increase of the cytoplasm was estimated in the rats fed with the low calcium diet for three and seven days. Enlargement of the cytoplasm and rather frequent mitoses of the chief cells were observed in the rats fed with the low calcium diet for 15 and 30 days. These findings suggest that the rapid bone loss in young rats induced by the low calcium diet is essentially due to stimulated activity of the parathyroid gland. The stimulated gland may be a result of hypertrophy at the early stage and a combination of hypertrophy and hyperplasia at the later stage of calcium deficiency. PMID- 11915357 TI - A morphological and immunohistochemical study of the ultimobranchial body in the Japanese lizard and the snake. AB - A morphological and immunohistochemical study of the ultimobranchial body of reptiles Japanese lizard and snake was carried out. The ultimobranchial body of the Japanese lizard was located adjacent to the left arch of the aorta between the trachea and esophagus. It was found as a cluster or group of cells with no capsule. Grimelius' silver impregnation and lead-hematoxylin staining produced positive reactions in some of the clustered cells and follicular cells. The same reaction pattern was observed with anti-calcitonin using the PAP method. The PAP reactions were positive to antiserum against pig calcitonin, but negative to antiserum against synthesized human calcitonin. Furthermore, the PAP reactions were negative to antiserum against tyrosine hydroxylase. The immunofluorescent study of the snake ultimobranchial body revealed that most of the clustered cells and some of the follicular cells were calcitonin-immunoreactive but none was tyrosine hydroxylase-immunoreactive. Certain histological similarities exist between the Japanese lizard ultimobranchial body and snake ultimobranchial body, but the distribution of calcitonin-positive cells were slightly different. In the Japanese lizard, the positive cells were scattered between the foliicles and the number was small. However, most of the cells which formed the cluster in the ultimobranchial body of snake were positive. The findings suggest that the configuration of amino acid in the Japanese lizard calcitonin and snake calcitonin are similar to that of pig calcitonin, and the reptile and the birds is a boundary of the tyrosine hydroxylase existence. PMID- 11915358 TI - A case of the horseshoe kidney. AB - During cadaver dissection for student education in anatomy laboratory of Gifu University School of Medicine, a case of the horseshoe kidney was observed in a 63-year-old Japanese female cadaver. The kidney was fused at the lower poles of the original kidneys by the parenchymatous isthmus and showed a typical horseshoe shape. The hili on both sides opened towards the ventral direction, and the ureters descended in front of the isthmus and entered the bladder normally. The location of the kidney was lower than that of the normal kidney. Four surplus arteries entered the isthmus and the lower part of the kidney besides the normal right and left renal arteries. The incidence of the horseshoe kidney during the dissecting practice at Gifu University School of Medicine from 1971 to 1997 was estimated to be 0.36% (4 out of 1130 bodies). PMID- 11915359 TI - Morphology of the dorsal lingual papillae in the newborn panther and Asian black bear. AB - The dorsal lingual surfaces of a newborn panther (Panthera pardus) and two newborn asian black bears (Selenarctos thibetanus) were examined by scanning electron microscopy (SEM). The tongues of the panther and asian black bear were about 40 mm in length and about 20 mm in width. Filiform, fungiform and vallate papillae were found. The filiform papillae were distributed over the entire dorsal surface of the tongue. In the panther, the filiform papillae on margin of the lingual apex were divided into two shapes which were horny or club-shaped papillae. The filiform papillae on the midportion were larger than those on the lateral region in size. The fungiform papillae also were divided into two shapes which were hemispherical or club-shaped papillae. In the asian black bear, the filiform papillae on the margin of the lingual apex were larger than those on margin of the panther tongue. The vallate papillae in the animals of two species were located on both sides of the posterior end of the lingual body. Each papilla was surrounded by a groove and crescent pad. PMID- 11915360 TI - Alteration of apoptosis in cleft palate formation and ectomesenchymal stem cells influenced by retinoic acid. AB - It has been shown that apoptosis is involved in normal embryonic development. The aim of the present study was to elucidate the role and alteration of apoptosis in the pathogenesis of cleft palate induced by retinoic acid (RA) and the ectomesenchymal stem (EMS) cells influenced by RA. RA was administered by gavage to pregnant C57BL/6N strain mice in the experimental group, and the control group received oil alone. Pregnant mice were killed at set periods of time thereafter and histologically analyzed. EMS cells explanted from the palatal shelves of embryonic mice were cultured and characterized by immunohistochemistry, growth curves and population-doubling time. The alterations of apoptosis of EMS cells and developing palatal shelves influenced by RA were evaluated by the terminal deoxynucleotidyl transferase-mediated UTP-biotin nick end-labeling (TUNEL) method. RA-treated mice showed formation of cleft palates resulted from the small size of the palatal shelves and their failure to lift. TUNEL staining showed that the number of apoptotic mesenchymal cells in palatal shelves in the RA-treated mice was increased significantly when compared with the control group. The primary culture of EMS cells proceeded successfully. The population-doubling time of RA-treated cells was much longer compared with non-treated EMS cells. RA also dramatically increased the number of apoptotic cells in EMS cells in vitro. We concluded that EMS cells are the crucial cells in palate development. RA could inhibit the proliferation and induced the apoptosis of EMS cells. The inhibition of growth and excess apoptosis of EMS cells may contribute to the formation of cleft palate and other orofacial congenital malformations. PMID- 11915361 TI - A scanning and transmission electron microscopic study of contractile trabecules in the rat spleen. AB - The fine structures of contractile trabecules in the splenic red pulp of the rat were examined by electron microscopy to elucidate their participation in the active contraction of the spleen. Numerous fine thready trabecules were developed in the red pulp. They were enveloped with a cytoplasmic layer of reticular cells and consisted of elongated smooth muscle cells, fascicles of collagenous fibrils and elastic fibers. Their fibrous components in the capsular ends extended in a triangular form of fanribs into the fibrous tunica of the capsule. Smooth muscle cell-like interstitial cells (SIC) were situated in the interfibrous spaces. Flattened SICs were affixed with cytoplasmic processes to the elastic lamina. The trabeculocapsular junctions were represented on the elastic lamina by grouped or isolated circular patches with concentrically arranged triangular processes and were also observed on the capsular serosa by plaques with scarce microvilli of serosal cells. Smooth muscle cells of the fine trabecules were equipped on the cell surface with anchoring structures to extracellular fibrous elements as previously described by Gabella (1981). Close associations were also seen between the smooth muscle cells and elastic fibers which were terminated to the fascicles of collagenous fibers. Cell-to-cell connections were expressed by fibrous connections between spiny processes and a small number of puntate intermediate junctions and nexuses. Unmyelinated nerve fibers with adrenergic terminals were seen in the intercellular spaces. We propose that for the rat spleen, the fine trabecules in the red pulp are muscular contractiles which are responsible for the active contraction due to sympathetic stimuli and the administration of alpha 1-adrenoceptor agonists, while the elastic lamina in the capsule plays a role in the comprehensive contraction of the subcapsular vascular bed. PMID- 11915362 TI - Fenestrated epithelium of the pelvic recess wall on the lateral parenchymal tissue in the dog kidney. AB - The epithelium of the pelvic recess separating the urine and the lateral parenchym of the dog kidney was examined by scanning and transmission electron microscopy (SEM and TEM) to try and identify the epithelial structures permitting filtration of urine substances. In cross sections through of the renal hilus, recesses were found in the pelvic space extending along the internal surface of the lateral parenchym. Light microscopy (LM) showed the external wall of the recess facing the parenchymal tissue to be a thin squamous epithelium underlined with a layer of smooth muscle cells. The cytoplasm of epithelial cells observed by TEM was extremely thin in comparison to the epithelia on other parts of the pelvic wall. Their peripheral cytoplasm usually demonstrated interdigitations or showed overlapping of cytoplasm of neighboring cells in two or three laminae. Distinct junctional structures were observed but no distension of the intercellular space was seen. The lumenal cell surface was relatively smooth, but the peripheral cytoplasm was equipped with either angular or stumped processes and deep cavae. In addition to a small number of cell organellae, numerous vesicles and vacuoles of varying sizes were conspicuously observed. The vacuoles contained flucculent materials and were fenestrated by pores closed by a diaphragm at fused potions of their limiting membrane with the adluminal and abluminal plasma membrane. The openings of the vacuoles were also seen at the basal cytoplasm. Similar fenestrations also occurred between the neighboring vacuoles in chains or conglomerations. Attenuated epithelial cytoplasm in close association with urinary tubules was also fenestrated by uniformly sized pores closed by thin diaphragm with punctuate central thickening. The fenestrations through the cytoplasm and between the chained vacuoles may thus be structures which have yet to be previously identified, and this finding seems to indicate that the filtration of urine substances occurs across the epithelial lining of the pelvic wall. Layers of thin smooth muscle cells with mutual connections are seen to underline the epithelium. The innermost muscle cells penetrated the basal lamina and thus come in contact with epithelial cells. However, their significance could not be clearly determined. PMID- 11915363 TI - Ultrastructure of collagen fibers and distribution of extracellular matrix in the temporomandibular disk of the human fetus and adult. AB - We quantitatively examined the distribution of these differences in extracellular matrices (collagen types I, III, and fibronectin) and elastic fibers under confocal laser scanning microscopy and electron scanning microscopy in terms of their contribution to the mechanics of the TMJ during development and in adults. Elastic fibers were found in the anterior and posterior bands in adults aged 40 years, and a few elastic fibers in the anterior band of the disk in adults aged 80 to 90 years. The extracellular matrix contents of the TMJ disk are shown in various detected levels in the anterior, intermediate, posterior bands of TMJ disk. During development, collagen fibers are arranged in a complex fashion from 28 weeks' gestation. These ultrastructures of the embryonic TMJ are resembled to that of adults aged the 40s, however the difference in extracellular matrix distribution found in embryonic stages and adults. They might reflect the differences in function between mastication and sucking or the changes in shape and form as results of functional disorders of the TMJ. PMID- 11915364 TI - Kupffer cell-mediated cytotoxicity induced by lipopolysaccharide0111:B4 is greater in dogs than in rats and monkeys. AB - To clarify the mechanism of sensitivity to an endotoxin lipopolysaccharide LPS0111:B4, which causes severe liver injury in a variety of animals, we have developed an in vitro assay to measure Kupffer cell-mediated cytotoxicity in the human liver cell line, WRL68. This assay could detect the decrease in Kupffer cell activity induced by gadolinium chloride (GdCl3), which is an inhibitor in Kupffer cells. Among Kupffer cells derived from dogs, rats, and monkeys, LPS activated canine Kupffer cells exhibited remarkably high cytotoxicity against WRL68 cells. This species difference is correlated with a species difference in the lethality of LPS0111:B4. Additionally, the conditioned medium of LPS activated canine Kupffer cells was also cytotoxic to WRL68 cells. To identify the mediators of this cytotoxicity, we measured the accelerated release of interleukin-1 beta, and interleukin-6 from Kupffer cells on stimulation with LPS0111:B4. From the correlation of the response to LPS0111:B4, interleukin-1 beta and interleukin-6 are considered to be responsible for the canine Kupffer cell-mediated cytotoxicity of LPS0111:B4. PMID- 11915365 TI - Evaluation of mitochondrial function and membrane integrity by dual fluorescent staining for assessment of sperm status in rats. AB - Dual fluorescent staining (DFS) with calcein acetoxy methyl ester (CAM), which labels the cellular esterase activity that is a major component of energy metabolism in cellular mitochondria, and with ethidium homodimer-1 (EthD-1) was used to evaluate mitochondrial function and membrane integrity in rat spermatozoa. The spermatozoa stained by DFS could be classified into three different populations microscopically when excited at 490 nm after 60 min incubation. 1) Spermatozoa, which were stained with CAM alone and had maintained either mitochondrial function or membrane integrity, were identified as live during incubation. 2) Spermatozoa, which were stained with EthD-1 alone and had lost either mitochondrial function or membrane integrity, were identified as already dead at the beginning of incubation. 3) Spermatozoa, which were stained with both CAM and EthD-1 and had maintained mitochondrial function with membrane breached, were identified as having died during incubation. Two toxicological tests, an in vitro triton X-100 experiment and an in vivo nitrobenzene experiment, were done. All spermatozoa were immobilized and lost either mitochondrial function or membrane integrity by 1.0% triton X-100 treatment. Almost no motile sperm were found at 0.1% in the triton X-100 group and in the groups treated with 60 and 40 mg/kg/day of nitrobenzene, and these spermatozoa maintained their mitochondrial function but had their membrane breached. In conclusion, the DFS procedure, which uses CAM and EthD-1, can clearly and visually identify the population of viable and dead spermatozoa simultaneously by fluorescence microscopy in rats. This is a useful technique to characterize sperm status, which is determined by the mitochondrial function assessed by CAM and membrane integrity evaluated by EthD-1. PMID- 11915366 TI - Effects of perinatal exposure to flutamide on sex hormones and androgen-dependent organs in F1 male rats. AB - Flutamide, which has antiandrogenic properties, was administered to pregnant rats, and effects on male offspring were examined. Crj: CD (SD) IGS (SPF) females were administered flutamide (0.15, 0.6, 2.5, 10.0, 100 mg/kg, p.o.) from gestation Day 14 to post parturition Day 3. The number of pups, body weights, clinical features, anogenital distance (AGD), nipple retention, testicular descent, and urogenital malformation in F1 males were examined. Hormone measurement, necropsy and histopathological examination were carried out at post neonatal Day 4 (PND 4) and PND 60. Sperm analysis was also carried out at PND 60. Decrease in body weight was seen in the 100 mg/kg group and the AGD was decreased at 2.5 mg/kg and above. Retention of nipples, hypospadia, vaginal pouches, penis malformation, unilateral ectopic testis, and decrease of organ weights (prostate, seminal vesicles, levator ani muscle plus bulbocavernosus muscle, testis) were observed at 10 mg/kg and above. Testicular testosterone (T) was increased significantly with 100 mg/kg at PND 4 and tendencies for increase were observed in serum T, LH and FSH at 10 mg/kg and more at the same time point. In contrast, elevated levels of LH and FSH were seen with 100 mg/kg at PND 60. Histopathological examination revealed defects or hypoplastic changes of genital organs (> or = 10 mg/kg), squamous metaplasia (10 mg/kg) or mucification (100 mg/kg) of the urethral diverticulum epithelium and inflammation of genital organs (100 mg/kg). Though only undescended testes lacked spermatogenesis at 10 mg/kg, atrophic change of seminiferous tubules and azoospermia were observed in the 100 mg/kg group, despite testicular descent. Perinatal administration of flutamide affected F1 male rats at 2.5 mg/kg and above. In addition to urogenital malformation, 100 mg/kg flutamide caused high LH and FSH levels at PND 60. This study indicates that the most sensitive parameter is AGD, whereby reduction was observed at 2.5 mg/kg. A clear no-effect level (NOEL: 0.6 mg/kg) was obtained in this perinatal study of an antiandrogenic chemical. PMID- 11915367 TI - Chronic toxicity in monkeys with the thiazolidinedione antidiabetic agent troglitazone. AB - The antidiabetic agent troglitazone was given to groups of 4 cynomolgus monkeys per sex at 300, 600, or 1200 mg/kg daily by gavage for 52 weeks. A group of 4 monkeys per sex received vehicle alone and served as controls. Emesis and soft stool or diarrhea occurred sporadically in all troglitazone-treated groups, but did not compromise animal health. There were no effects on body weight or food consumption, or ophthalmologic, electrocardiographic, or echocardiographic parameters. Erythrocyte count, hemoglobin, and hematocrit decreased 8% to 16% in males at all doses and serum cholesterol decreased 30% to 46% in both sexes at all doses. Urinary ketones were increased in several animals at 600 and 1200 mg/kg. Absolute and relative liver weights increased at all doses in both sexes by 40% to 71%. The only microscopic change attributable to troglitazone treatment was minimal to mild bile duct hyperplasia in males at all doses and in females at 600 and 1200 mg/kg. No differences in systemic exposure were apparent between sexes. Over the dose range tested, AUC(0-24) values were 27 to 102 micrograms.hr/ml of troglitazone, 401 to 2060 micrograms.hr/ml of its sulfate conjugate, and 34 to 312 micrograms.hr/ml of its quinone metabolite. Therefore, oral administration of troglitazone to monkeys at 300, 600, and 1200 mg/kg for 52 weeks resulted in significant systemic exposure, with only minimal gastrointestinal, hematologic, and hepatic effects. PMID- 11915368 TI - Endocrinological assessment of toxic effects on the male reproductive system in rats treated with 5-fluorouracil for 2 or 4 weeks. AB - Endocrinological assessment of male reproductive toxicity was carried out in SD Slc male rats treated with 5-FU (0, 20, 30 mg/kg/day) orally for 2-week or 4-week term. Serum hormone levels including GnRH, FSH, LH, prolactin, total and free testosterone, inhibin B, pro-alpha C, and activin A were determined as well as histopathological examination of the reproductive organs. The 5-FU treated groups showed histopathological changes in the testis such as degeneration of seminiferous epithelium. An obvious decrease in serum testosterone level was observed with a reduced organ weight of the seminal vesicle and prostate. However, no significant changes were noted in serum LH or FSH levels, nor in the morphological examination of the Leydig cells. Decreased serum levels were noted in activin A and prolactin. An increased serum level was noted in GnRH and pro alpha C whose synthesis is regulated by FSH. Serum inhibin B levels showed a tendency toward decreasing with morphological change (vacuolation) in Sertoli cells. These results indicated that male reproductive toxicity induced by 5-FU would be augmented by decreased serum prolactin and testosterone levels as well as a decreased function of Sertoli cell, in addition to the direct cytotoxic effects on germ cells. It is suggested that these endocrinological changes related to male reproductive toxicities can be detected even in the 2-week treated study. PMID- 11915370 TI - [Two cases of retinal detachment following laser in Situ keratomileusis repaired by scleral buckling surgery]. AB - BACKGROUND: As laser in situ keratomileusis (LASIK) becomes the treatment of choice in the field of refractive surgery, postoperative rhegmatogenous retinal detachment has also begun to occur. However, since it has not been reported yet here in Japan, we report two cases of retinal detachment following LASIK with suggestions based on our experience. CASES: A 34-year-old male(Case 1) and a 26 year-old male(Case 2) suffered from retinal detachment one month and eight months after LASIK, respectively. Contributing retinal breaks were a horseshoe tear in Case 1 and atrophic holes in the lattice degeneration in Case 2. Each patient underwent partial scleral buckling with no intraoperative complication, resulting in successful retinal reattachment. Postoperatively, myopic changes developed and persisted one month in Case 1 and several months in Case 2. CONCLUSIONS: Great attention should be paid to the possibility of postoperative myopia after a repair of retinal detachment following LASIK. Thus, we stress the necessity of close cooperation between LASIK surgeons and vitreoretinal surgeons to settle the matter. PMID- 11915369 TI - Prevention by natural food anthocyanins, purple sweet potato color and red cabbage color, of 2-amino-1-methyl-6-phenylimidazo[4,5-b]pyridine (PhIP) associated colorectal carcinogenesis in rats initiated with 1,2 dimethylhydrazine. AB - The potential of purple sweet potato color (PSPC) and red cabbage color (RCC), natural anthocyanin food colors, to modify colorectal carcinogenesis was investigated in male F344/DuCrj rats, initially treated with 1,2 dimethylhydrazine (DMH) and receiving 2-amino-1-methyl-6-phenylimidazo[4,5 b]pyridine (PhIP) in the diet. After DMH initiation, PSPC and RCC were given at a dietary level of 5.0% in combination with 0.02% PhIP until week 36. No PSPC or RCC-treatment-related changes in clinical signs and body weight were found. Incidences and multiplicities of colorectal adenomas and carcinomas in rats initiated with DMH were clearly increased by PhIP. In contrast, lesion development was suppressed by RCC, or tended to be inhibited by PSPC administration. Furthermore, in the non-DMH initiation groups, induction of aberrant crypt foci (ACF) by PhIP was significantly decreased by RCC supplementation. The results thus demonstrate that while PhIP clearly exerts promoting effects on DMH-induced colorectal carcinogenesis, these can be reduced by 5.0% PSPC or 5.0% RCC in a diet under the present experimental conditions. PMID- 11915371 TI - [Light scattering observed on the surface of acrylic intraocular lenses ten years after implantation]. AB - PURPOSE: To report the long-term status of acrylic intra-ocular lenses. CASES: We describe two cases of increased light scattering on the surface of acrylic intraocular lenses ten years after implantation. The scattering was non symptomatic and visual acuity was normal. Biomicroscopy and a Scheimpflug photography system showed that the scattering apparently derived from the anterior surface of the lens, but no deposit or membranous structure was observed on the lens surface. CONCLUSION: Surface scattering from acrylic intra-ocular lenses is a new finding in long-term observations. A possible etiology may be structural changes of the internal polymer. PMID- 11915372 TI - [Experimental study of glistening in silicone intraocular lenses]. AB - PURPOSE: To assess the whether glistening(GP) occurs on silicon lenses. METHODS: ALLERGAN, SI-40NB and CANON STAAR, AQ110NV lenses(Alcon MA60BM control). The lenses were immersed in 50 degrees C physiological saline solution for 2 hours, and then immersed in 35 degrees C physiological saline solution. Changes in the lens were observed regularly with a slit lamp microscope until the 90th day. RESULTS: Numerous microvacuoles of 10 microns diameter occurred in the MA60BM and of several microns in the SI40NB. Some clouding was seen in the AQ110 NV. CONCLUSION: The presence of numerous microvacuoles in the SI40NB means there may be GP. The light clouding in the AQ110NV that was resolved with drying is thought to be due to particles in the material which cause discoloration. PMID- 11915373 TI - The World Trade Center disaster: a brief on-site report to the Nippon Ganka Gakkai Zasshi. PMID- 11915374 TI - [Ideal designs, materials and functions for intraocular lens]. PMID- 11915375 TI - [The concept of induced components in multifocal electroretinograms]. AB - PURPOSE: In the late period of a multifocal electroretinogram (mfERG) waveform, higher order-kernels copy themselves. These images are called projection components(or induced components), and contribute to the particular waveform of mfERG. I describe this feature, which the developer of the multifocal technique was fully aware of. METHOD: Inter-kernel comparisons were made of mfERGs, focusing on the characteristics and influence of induced components. RESULTS AND DISCUSSION: Even though induced components have never been described or discussed in detail in Japan, they are certainly one of the key elements in learning how the multifocal technique works and what the difference is between multifocal and conventional ERGs. PMID- 11915376 TI - [Changes in filtering bleb with laser treatment after non-penetrating trabeculectomy]. AB - PURPOSE: Ultrasound biomicroscopy(UBM) was performed on filtering blebs with laser treatment including laser gonioplasty(GP), laser iridotomy(LI), and YAG laser trabeculopuncture(YLT) after non-penetrating trabeculectomy(NPT). The filtering blebs were grouped into four types, L, H, E and F, and the spaces under the scleral flap were classified into three. La, S, and N. CASES: In case 1, an F type, an S bleb was not changed despite laser treatment 9 months after NPT. In case 2, YLT was performed twice to release the incarcerated iris. The filtering bleb was changed to the L type, an La bleb. In case 3, an F type, an S bleb was turned into an H type after GP, LI, and YLT. In case 4, GP effectively released peripheral anterior synechia formation and the filtering bleb was kept as an L type, La. CONCLUSION: Appropriate laser treatment may be effective in turning a flattened and localized bleb into a good filtering bleb. PMID- 11915377 TI - [Repair of canalicular lacerations using silicone tubes(80 cases)]. AB - PURPOSE: Long-term canalicular intubation is preferred for improving the prognosis of canalicular lacerations. We have improved the intubation step by step from methods I to V. The present paper describes the modifications that we previously made as well as the prognosis of each method. SUBJECTS AND METHODS: During the past 13 years, 80 cases of canalicular lacerations underwent the intubation with silicone tubes in our department. The methods used are designated by I (placed in the affected side), II(ring-form intubation into the lacrimal sac), III(loop-form intubation into the nasal cavity), IV(Fayet's tube with a punctal plug is placed in the affected side), and V(a KAWAI open tube with a punctal plug is placed in the affected side). RESULTS: Each of the canalicular intubations improved the prognosis: the ratio of patients showing improvement was 85.7% for method I, 94.7% for II, 97.3% for III, and 100% for IV and V. Methods IV and V caused fewer complications during long-term intubation. CONCLUSION: Method V that we developed permits tear drainage, and decreases epiphora. The punctal plug prevents the tubing from dropping out, and therefore permits longer placing of the canalicular tubing than does method II or III. Thus, our method V seems to improve the prognosis of canalicular laceration repairing. PMID- 11915379 TI - [A case of granulomatous panuveitis associated with multiple sclerosis]. AB - BACKGROUND: Uveitis is not often reported in patients with multiple sclerosis(MS) in Japan, but retinal vasculitis and pars planitis are common findings in uveitis patients with MS in the U.S. and European countries. CASE: A 50-year-old man developed bilateral granulomatous panuveitis 10 years after initial onset of MS. FINDINGS: Examination showed bilateral anterior uveitis with mutton fat keratic precipitate and retinal vasculitis for 360 degrees. The anterior uveitis responded well to topical corticosteroid but the retinal vasculitis did not. The possibility of sarcoidosis, tuberculosis, and other diseases associated with granulomatous uveitis were excluded by systemic examinations. CONCLUSION: We encountered a rare case of granulomatous panuveitis associated with MS. PMID- 11915380 TI - Ambulance services. Get your kit off. AB - An ambulance trust with a deficit of 1 m Pounds in 2000 and poor response times has balanced its books and exceeded response-time targets. A hierarchical, military culture has been changed into a more participative one. Union representatives now sit on the interviewing board for management jobs. These changes have allowed the trust to become more integrated with the NHS as a whole. PMID- 11915378 TI - [Clinical dose finding study of indocyanine green to detect choroidal neovascularization in patients with exudative age-related macular degeneration]. AB - PURPOSE: To evaluate the proper clinical dosage of indocyanine green(ICG) in angiography for detecting choroidal neovascularization(CNV) of exudative age related macular degeneration(AMD). SUBJECTS AND METHODS: Indocyanine green angiography (IA) was performed using a randomized crossover method with two different doses on two occasions. Ease of detection, side effects and clinical serum data were also evaluated. RESULTS: Among the 39 eyes, detection of CNV using 12.5 mg and 25 mg was most effective in 21 and 31 eyes respectively, showing a statistically significant difference. Slight vomiting was observed temporarily in one patient who had taken 25 mg. CONCLUSION: A dose of 25 mg is appropriate for detection of CNV of exudative AMD and this dosage raises no safety concerns. PMID- 11915382 TI - Open space. Falling on deaf ears. PMID- 11915383 TI - Training centre for health records technology and medical records department. PMID- 11915384 TI - Do not exceed the stated dose (a prescription for electronic medical records). PMID- 11915385 TI - A code of ethics for health informatics professionals. PMID- 11915387 TI - Geriatric pharmacotherapy: problems with adherence. AB - Because of their many health problems, older people are likely to receive a greater quantity and variety of medications. Every new drug added to a medication schedule will compound the risk of adverse reactions for the older population, who manage drugs less efficiently due to their altered body status, drug therapy becomes further complicated. To minimise the risks associated with drug therapy and ensure that medications do not create more problems than they solve, close supervision, education and adherence of drug usage in older adults are essential. PMID- 11915388 TI - 'The experience of community mental health case management provided from an acute in-patient psychiatric unit'. AB - In Australia, case management is the cornerstone of mental health service delivery for seriously ill clients living in the community. In this study, case management was provided from an acute, inpatient psychiatric unit; a model thought to be unique. Findings from this qualitative study explicated the experience of case management from client and case manager (CM) perspectives. They note the nature, purpose, processes and outcomes of case management within that context. Findings were positive, suggesting clients and CM's develop a therapeutic alliance through which interventions are implemented and which result in clients experiencing personal (re) integration and enhanced well-being. These findings are discussed and they suggest an alternative model of service delivery well regarded by both clients and CM's. PMID- 11915389 TI - Crown cementation and desensitization. PMID- 11915390 TI - Greening of the tooth-amalgam interface during extended 10% carbamide peroxide bleaching of tetracycline-stained teeth: a case report. AB - At-home bleaching with 10% carbamide peroxide in a custom-fitted tray has been shown to have some minor effects on certain brands of amalgam, pertaining to mercury release, but generally, effects on amalgam are not considered clinically significant. However, in this case report, a greening of the tooth structure in certain areas immediately adjacent to amalgam restorations in the maxillary and mandibular first molars occurred during tooth whitening. Other amalgam restorations in mandibular and maxillary second molars in the same mouth did not demonstrate any green discoloration of the teeth. Upon removal of the affected amalgam restorations, recurrent decay was present in the areas of tooth greening but not in other areas adjacent to the restoration. The teeth were restored with posterior composite restorations. Whether the green discoloration was a result of some loss of material from a particular brand of amalgam, indicating leakage, or indicative of original or recurrent tooth decay is unclear in this single-patient situation. Other patients in the same study did not demonstrate this occurrence. Dentists should be ready to replace amalgam restorations should this green discoloration in adjacent tooth structure occur during bleaching, in case decay is present. CLINICAL SIGNIFICANCE: The unusual discoloration cited suggests that amalgam restorations in potentially esthetic areas, including the lingual of anterior teeth, should be replaced prior to bleaching, to avoid the problem of difficult stain removal or translucency allowing restoration visibility following bleaching. PMID- 11915391 TI - Esthetic correction of gingival recession using a modified tunnel technique and an acellular dermal connective tissue allograft. AB - Esthetic correction of gingival recession is an important goal of periodontal therapy. This article describes a surgical technique that combines a modified tunnel technique and an acellular dermal connective tissue allograft. With the aid of vertical incisions, a tunnel is created under the buccal mucosa of the affected tooth. These incisions enable easy access for graft placement and create mobility for gingival coronal positioning. The use of an acellular dermal connective tissue allograft eliminates the need for a surgical palatal donor site. This minimizes postsurgical complications. CLINICAL SIGNIFICANCE: The combination of a modified tunnel technique and an acellular dermal connective tissue allograft permits esthetic root coverage in a manner that reduces postsurgical complications. PMID- 11915392 TI - Effect of bleaching agents on the hardness and morphology of enamel. AB - PURPOSE: The goal of the present laboratory study was to analyze the effect of two home bleaching agents (Opalescence 10%, Ultradent, and Hi-Lite II, Shofu) as well as the separate solutions of 10% carbamide peroxide primary components (3% hydrogen peroxide and 7% urea) on enamel microhardness and surface morphology. MATERIALS AND METHODS: Thirty human molars stored in water for no longer than 3 months were selected. Their facial and lingual surfaces were embedded in acrylic resin, and the specimens were polished with up to 600-grit sandpaper. The 30 molar sections were divided into five groups as per treatment modality (n = 6); group 1, 10% carbamide peroxide bleach with carbopol (Opalescence, Ultradent); group 2, oxygen-free gel bleach (Hydroxylite, Hi-Lite II, Shofu); group 3, 3% hydrogen peroxide with carbopol (Dermus Pharmacy, Florianopolis, SC, Brazil); group 4, 7% urea treatment (Dermus Pharmacy); and control, group 5, artificial saliva (Dermus Pharmacy). Prior to treatment, six indentations (Vickers) were made on each of the six surfaces in each group under a load of 100 g for 30 seconds. Bleaching procedure was conducted for 3 hours per day for 2 weeks, the control group excepted. Following each bleaching session, the specimens were washed with deionized water and kept in artificial saliva at 37 degrees C, this solution being changed daily. Pre- and post-treatment microhardness data were analyzed using a two-way analysis of variance. In a similar way, 30 other specimens underwent the same procedures. After the bleaching period, the samples were observed under the scanning electron microscope. RESULTS: Specimens submitted to bleaching treatment with 3% hydrogen peroxide gel as described in this study showed a significant reduction of surface microhardness (p < .0001). The other treatment modalities had no effect on surface microhardness. No morphologic alterations were observed on enamel submitted to bleaching gel of 10% carbamide peroxide (Opalescence), oxygen-free gel (Hi-Lite II, Shofu), or 7% urea solution compared with the control (artificial saliva). However, the specimens bleached for 3 hours per day for 2 weeks with 3% hydrogen peroxide gel presented areas of mild erosion. The effect, nevertheless, was not uniform, occurring with varying intensity on all samples of enamel bleached with 3% hydrogen peroxide. CLINICAL SIGNIFICANCE: Bleaching treatment conducted with two home bleaching agents (Opalescence 10%, Ultradent, and Hi-Lite II, Shofu) had no adverse effects on enamel microhardness or on surface morphology. Bleaching with 3% hydrogen peroxide can have a negative effect on enamel hardness and surface morphology when performed as described in this study. PMID- 11915393 TI - In vivo influence of residual moisture on microtensile bond strengths of one bottle adhesives. AB - PURPOSE: This study evaluated the microtensile bond strengths of three dentin adhesives applied on clinically moist dentin or on dentin that was dried with air for 5 seconds. The null hypothesis to test was that the level of residual moisture does not influence bond strengths when restorations are placed in vivo. MATERIALS AND METHODS: Twenty-four premolars scheduled to be extracted for orthodontic reasons from patients between the ages of 15 and 23 years were restored with one of the following adhesive systems followed by a mini hybrid composite resin: Excite (Ivoclar/Vivadent), an ethanol-based dentin adhesive; Prime & Bond NT (Dentsply/Caulk), an acetone-based dentin adhesive; and Single Bond (3M ESPE), an ethanol and water-based dentin adhesive. After extraction, the specimens were sectioned with a slow-speed diamond saw in two perpendicular directions to obtain sticks with a cross-section of 0.7 +/- 0.2 mm2. The specimens were attached to a Geraldeli device and fractured using a universal testing machine at a crosshead speed of 1 mm per minute. RESULTS: For each dentin adhesive, there were no statistical differences between means for dry dentin versus moist dentin. Single Bond and Prime & Bond NT ranked in the same statistical subset regardless of the moisture condition of the substrate. Both Excite, dry, and Excite, moist, resulted in statistically lower bond strengths than Single Bond, moist, but similar to those of Single Bond, dry, Prime & Bond NT, moist, and Prime & Bond NT, dry. CLINICAL SIGNIFICANCE: In this study, the level of residual moisture did not influence microtensile bond strengths. Clinically, the degree of moisture left on the dentin surface upon rinsing off the etching gel may not be as relevant as previously reported in laboratory studies. PMID- 11915395 TI - Shear bond strength of bis-acryl composite provisional material repaired with flowable composite. AB - BACKGROUND: Time and expense are considerable when refabrication of a bis-acryl composite provisional prosthesis is required, so an effective method of repair is desirable. Attempts at self-repair have been ineffective, and no reports of repair specific to bis-acryl composites were found in the literature. PURPOSE: To investigate the strength of a bis-acryl composite repair using flowable composite after two surface treatments and three storage conditions. MATERIALS AND METHODS: Thirty specimens of bis-acryl composite were made and divided into six groups. Half of the specimens received air abrasion only, and half received air abrasion followed by application of an intermediate bonding resin. A flowable composite was then applied to all specimens. Following three different storage conditions, the flowable composite was debonded using a universal testing machine in tensile mode. RESULTS: All specimens failed cohesively within the bis-acryl composite rather than at the repair interface. Surface treatment with air abrasion alone resulted in significantly higher shear bond strength values than when air abrasion and intermediate bonding resin were combined. Bond strength values were significantly higher in thermocycled specimens than in air-dried specimens. CONCLUSIONS: Use of air abrasion followed by application of flowable composite proved to be a successful technique for bis-acryl composite repair. CLINICAL SIGNIFICANCE: Repair of bis-acryl composite provisional material with a flowable composite is effective, expedient, and inexpensive. PMID- 11915394 TI - A computer-controlled anesthetic delivery system in a periodontal practice: patient satisfaction and acceptance. AB - BACKGROUND: Many dental patients are fearful about receiving intraoral injections of local anesthetic. In fact, many patients cite injections as their primary reason for avoiding dental treatment. In late 1997, however, a computerized local anesthetic delivery system was introduced to address the problems of pain and anxiety associated with these injections. The author initiated a year-long assessment study in which patients evaluated and rated injections administered with the computerized device as compared with previously administered standard injections. METHODS: One hundred and fifty randomly selected patients received local anesthesia delivered via the computer-controlled anesthetic delivery system. All patients had at some time received standard injections during treatment at the practice. Different types of injections were administered to all quadrants, enabling the researcher to qualify results according to injection type and location. Patient responses were recorded and evaluated to assess the levels of anxiety and pain associated with the computer-driven system. RESULTS: Overall, 71.4% of patients reported the experience to be superlative, positive, or somewhat positive. Results indicated that the system was highly preferred to traditional injection techniques, regardless of the arch and quadrant receiving the injection. Several factors may have contributed to this preference, including increased patient comfort, the nonthreatening appearance of the instrument, and the lack of residual facial numbness commonly associated with oral anesthetic injections. CLINICAL SIGNIFICANCE: By removing the discomfort associated with the dental injection, which is a common source of patient anxiety, use of the computer-controlled anesthetic delivery system may encourage patient acceptance of recommended periodontal treatment modalities. PMID- 11915396 TI - Endodontic technology offers the restorative dentist reliability. PMID- 11915397 TI - Porcelain veneers. PMID- 11915399 TI - Knowledge of the lymphatics of the head & neck is essential to the accurate examination of lymph nodes. PMID- 11915400 TI - [Prevalence of ulcus cruris in home care nursing. An epidemiological study in North Rhine-Westphalia]. AB - The aim of this study was to provide reliable data on prevalence of leg ulcers in home-care setting. A descriptive explorative cross-sectional design was used. In March 2000, a randomised sample of 520 community home nursing services in North Rhine Westphalia was sent a standardised questionnaire. Apart from questions about the type of service provider and its geographical population, demographic patient data and information about duration and cause of ulcer was collected. The findings based on the data gathered from 161 nursing home-care services caring for a total 12,156 patients. The overall prevalence was 2.68% (+/- 0.29%) for leg ulcers in home-care setting. The median age of the 326 affected patients was 77.5 years. Women are more than twice as often affected as men, which could be due to the longer life-expectancy for women. A major problem for home nursing services, however, is the large number of leg ulcers of unknown origin. This was the case in almost a quarter of the sample. Depending on the causal factors, compression therapy is of paramount importance and crucial to the successful outcome of the treatment of the leg ulcer. The markedly chronic nature of leg ulcers is again confirmed by this study. Currently, leg ulcers have an average duration of ten months. However, it is not unusual for affected patients to suffer for years or even decades. Almost 80% of patients had their ulcer dressed once a day by the nursing home-care service. PMID- 11915398 TI - The clinical effects of saline and aloe vera rinses on periodontal surgical sites. PMID- 11915402 TI - [Publishing in "Pflege" journal]. PMID- 11915403 TI - ["... but it is for many also a large step: from the brink of death to the celebration of life...". Routine care with new drug treatment possibilities for HIV positive and AIDS patients from the viewpoint of patients]. AB - New combination therapy provides a stabilization of health and a longer life expectancy for people living with HIV/AIDS who are able to deal with their drugs and the strict daily medical regime inflicted. This of course produces high expectations. Due to the success of the combination therapy, the psychosocial problems associated with it have been pushed to the background. How do people living with HIV/AIDS evaluate the changes that occurred since the introduction of the new combination therapy? Which changes affect their daily routine most? Is there a need for change in nursing care? In order to address these questions, this study attempts to grasp and analyse the changes and coping strategies of people that are treated with combination therapy at source. The results are discussed taking a perspective of positive change and support. The main aim of this study was to contribute to a better understanding of people living with HIV/AIDS that are treated with combination therapy and to improve their professional support. PMID- 11915401 TI - ["Nursing care visit: possibilities and limits". An empirical study in Bremen, Hamburg, Mecklenburg-Vorpommern, Lower Saxony and Schleswig-Holstein]. AB - This study examines the distribution, understanding and objectives of nursing visits as well as the implementation and the experience with nursing visits in 87 hospitals in North Germany (Bremen, Lower Saxony, Hamburg, Schleswig-Holstein and Mecklenburg-Western Pomerania). The survey was conducted using a questionnaire. It was found that nursing visits are implemented in 31% of the departments. By means of the nursing visit the patients are involved in the nursing process. The objectives of nursing visits are patient orientation. and the improvement of the nursing quality. The nursing visit is criticised because of both the level of organisation and the time spent on nursing visits as well as the lack of training, maintaining confidentiality and questions of data protection. The study contributes to a better understanding of nursing visits and a clearer definition of the term. It is the first empirical study of this type that has been conducted in Germany. PMID- 11915404 TI - NHS politics. Winging it. AB - At present, NHS managers are highly constrained, suffering excessive regulation and central control. More autonomy for trusts would mean fewer directives and less performance management. Giving trusts a new organisational form, such as a public interest company or foundation hospital, might be reinvigorating and would not involve further reorganisation. These new freedoms should be accompanied by new accountabilities, not solely to politicians but to independent NHS regulators, local communities and patients. Devolved power and greater patient choice could produce a more responsive NHS. Its potential needs to be explored through experimentation and evaluation. PMID- 11915405 TI - Primary care trusts. Ready, steady, stop. AB - A survey of primary care groups and trusts in South West region found most felt they had little autonomy. Most were level-2 PCGs, taking responsibility for managing a budget for commissioning services. About a third were still relying heavily on their health authority for commissioning. The results suggest that the proposal to allocate 75 per cent of NHS funds to PCTs by 2004 will be unrealistic in some areas. PMID- 11915406 TI - Life course transitions and natural disaster: marriage, birth, and divorce following Hurricane Hugo. AB - Change in marriage, birth, and divorce rates following Hurricane Hugo in 1989 were examined prospectively from 1975 to 1997 for all counties in South Carolina. Stress research and research on economic circumstances suggested that marriages and births would decline and divorces would increase in affected counties after the hurricane. Attachment theory suggested that marriages and births would increase and divorces would decline after the hurricane. Time-series analysis indicated that the year following the hurricane, marriage, birth, and divorce rates increased in the 24 counties declared disaster areas compared with the 22 other counties in the state. Taken together, the results suggested that a life threatening event motivated people to take significant action in their close relationships that altered their life course. PMID- 11915407 TI - Marital quality, coping with conflict, marital complaints, and affection in couples with a depressed wife. AB - This study compared three groups of women--outpatient depressed, inpatient depressed, and community control--and their husbands on a range of variables including marital functioning and styles of coping with conflict. Outpatient depressed couples reported greater marital distress and more destructive and less constructive tactics for resolving conflict than did community control couples. They also were more likely to have been previously married and to express regrets about having married their current husbands. There were smaller and less consistent differences for couples with inpatient depressed spouses, although inpatient couples with younger wives were similar to outpatient depressed couples. Both groups of depressed women and their husbands reported fewer expressions of affection and more complaints about the marriage than did control couples. Results are discussed in terms of interpersonal perspectives on depression. PMID- 11915408 TI - Couples coping with congestive heart failure: role and gender differences in psychological distress. AB - Psychological distress and marital quality were assessed with male (n = 128) and female (n = 49) congestive-heart-failure (CHF) patients and their spouses. Hopkins Symptom Check List--25 scores were in the distressed range for 57% of patients and 40% of spouses. This role difference was greater for men than for women, and a gender difference (more distress in women than men) was greater for spouses than for patients. The patient's distress, but not the spouse's, reflected the severity of the patient's illness, and distress for both partners correlated negatively with ratings of marital quality. Female-patient couples reported better relationship quality than male-patient couples, however, and a mediation analysis indicated that the gender difference in spouse distress could be explained by marital quality. Results highlight the contextual nature of CHF distress and suggest that role differences in distress vary by gender. PMID- 11915409 TI - Pathways to adversity in young adulthood among early childbearers. AB - Women who give birth as teens differ from those who delay childbearing before and after a birth. These preexisting differences may account for the adverse outcomes faced by early childbearers in young adulthood. This study tested whether a history of conduct disorder, low IQ and educational attainment, and low childhood socioeconomic status accounted for poor psychosocial adjustment at age 26 among early childbearers. Study members were 482 women in a birth cohort, 26% of whom had given birth by age 26 in 1999. Findings supported the hypothesis that individual and family background factors partially accounted for the adverse socioeconomic, mental health, and interpersonal outcomes faced by young mothers. However, early childbearing exacerbated the difficulties associated with these risk factors. PMID- 11915410 TI - Sibling conflict in middle childhood predicts children's adjustment in early adolescence. AB - Associations between sibling conflict in middle childhood and psychological adjustment in early adolescence were studied in a sample of 80 boys and 56 girls. Parents and children provided self-report data about family relationships and children's adjustment. Parents' hostility to children was assessed from videotaped interactions. Results showed that sibling conflict at Time 1 predicted increases in children's anxiety, depressed mood, and delinquent behavior 2 years later. Moreover, earlier sibling conflict at Time 1 accounted for unique variance in young adolescents' Time 2 anxiety, depressed mood, and delinquent behavior above and beyond the variance explained by earlier maternal hostility and marital conflict. Children's adjustment at Time 1 did not predict sibling conflict at Time 2. Results highlight the unique significance of the earlier sibling relationship for young adolescents' psychological adjustment. PMID- 11915411 TI - Parental management of adolescent peer relationships in context: the role of parenting style. AB - Three hundred 9th-graders and their best friends participated in this short-term longitudinal study. Confirmatory factor analysis of the Parental Management of Peers Inventory (N. Mounts, 2000) suggested that the 4-factor structure provided a good fit to the data. Significant differences in adolescents' perceptions of parental management of peers occurred across 4 parenting style groups. Structural equation modeling was used to examine the relation among 5 aspects of parental management of peers and adolescents' Time 1 drug use, friends' Time 1 drug use, and adolescents' Time 2 drug use. The parental management styles of monitoring, guiding, prohibiting, and supporting all had significant paths in the model, whereas neutrality was not significant. Multiple group comparisons were used to examine whether parenting style moderates the relation between parental management of peers and the drug use outcomes. Parenting style functioned as a moderator for 7 of the paths in the model. PMID- 11915412 TI - Predictors of self-esteem for Mexican American and European American youths: a reexamination of the influence of parenting. AB - Decades of research with European American middle-class families have found significant relations between parenting behavior and child self-esteem. Similar research with minority and low-income families is rare. The present study examined the relation between parenting practices and child self-esteem among 70 Mexican American and 161 European American youths. The analyses consisted of regressing child self-esteem on parenting practices (acceptance, rejection, inconsistent discipline, and hostile control), ethnicity, socioeconomic status (SES), and the interactions between ethnicity, SES, and parenting practices. Several main effects and interactions were significant; for each interaction, behavior of low-income or Mexican American parents had less influence on children's self-esteem than did similar behavior by middle-class or European American parents. PMID- 11915413 TI - Instruction by mothers and fathers during problem solving with their young children. AB - Mothers and fathers of 163 5-year-olds were observed interacting with their children in dyads on 2 separate occasions on a familiar and unfamiliar cognitive activity. Within- and between-family comparisons were conducted. Few differences in the instruction provided by mothers and fathers appeared, and those that did were on the unfamiliar task. On this task, instruction by mothers, within and across families, was more responsive to children's changing skill than was instruction by fathers. Directive and disapproving comments by parents were related to poorer posttest performance by the child. High level of instruction by both parents was related to better posttest performance than was low level of instruction by parents. Contributions of parents to their children's cognitive development are discussed. PMID- 11915414 TI - Child adjustment in joint-custody versus sole-custody arrangements: a meta analytic review. AB - The author meta-analyzed studies comparing child adjustment in joint physical or joint legal custody with sole-custody settings, including comparisons with paternal custody and intact families where possible. Children in joint physical or legal custody were better adjusted than children in sole-custody settings, but no different from those in intact families. More positive adjustment of joint custody children held for separate comparisons of general adjustment, family relationships, self-esteem, emotional and behavioral adjustment, and divorce specific adjustment. Joint-custody parents reported less current and past conflict than did sole-custody parents, but this did not explain the better adjustment of joint-custody children. The results are consistent with the hypothesis that joint custody can be advantageous for children in some cases, possibly by facilitating ongoing positive involvement with both parents. PMID- 11915415 TI - Chiropractic for midwives. An introduction. PMID- 11915416 TI - Self-hypnosis in midwifery. PMID- 11915417 TI - Tackling birth trauma with cranio-sacral therapy. PMID- 11915418 TI - Massage for childbirth and pregnancy--8 years on. PMID- 11915419 TI - Complementary therapies in midwifery. Establishing a database. PMID- 11915420 TI - Give dads a chance! Baby massage for fathers. PMID- 11915421 TI - Building and restoring confidence. PMID- 11915423 TI - The Guthrie test. Midwifery basics. PMID- 11915424 TI - What's it like to work in ... Zimbabwe. PMID- 11915422 TI - Optimal duration of exclusive breastfeeding. PMID- 11915425 TI - Action: promoting normal birth. PMID- 11915426 TI - Complementary therapies in the midwifery curriculum. PMID- 11915427 TI - ABO incompatibility. PMID- 11915428 TI - Epilepsy drugs advice. PMID- 11915429 TI - Home and hospital births equally safe. PMID- 11915430 TI - Cot death research. PMID- 11915431 TI - Language in ageing persons with Down syndrome. AB - Several cross-sectional studies and one longitudinal study were conducted on the language abilities of various cohorts of persons with Down syndrome aged between 14 and 50 years. No significant difference was observed on any of the receptive and productive morphosyntactic and lexical measures used, suggesting no marked change in the language of these persons from adolescence onto late adulthood. Repeated measures of cerebral metabolic rate (CMR) for fluorodeoxyglucose using a Positron Emission Tomography were made over a 4-year interval with 7 participants with Down syndrome aged between 37 and 49 years. A gradual decrease in global CMR for both cerebral hemispheres and for each participant was documented. It was particularly marked for 3 participants. However, no language deterioration could be associated with their marked lowering in CMR. PMID- 11915432 TI - Phonological awareness in children with Down syndrome. AB - Research in the area of phonological awareness has mainly focused on the nature of the relationship between reading ability and awareness of phonemes. However, a recent study of phonological awareness in children with Down syndrome questioned the existence of any necessary relationship (Cossu, Rossini & Marshall, 1993). This paper describes a study of phonological awareness in children with Down syndrome with varying levels of reading ability. The sample consisted of 10 male and 7 female children with Down syndrome (aged 9 years 2 months to 14 years 5 months). All children received a battery of tests which consisted of assessments of: 1) phonological awareness, 2) reading and spelling competence, 3) non-word reading and spelling ability, and 4) non-verbal measures. Children with Down syndrome demonstrated measurable levels of phonological awareness. Significant positive correlations were found between phonological awareness and: reading and spelling competence, ability to spell non-words and non-verbal measures. PMID- 11915433 TI - Improving the classroom listening skills of children with Down syndrome by using sound-field amplification. AB - Many children with Down syndrome have fluctuating conductive hearing losses further reducing their speech, language and academic development. It is within the school environment where access to auditory information is crucial that many children with Down syndrome are especially disadvantaged. Conductive hearing impairment which is often fluctuating and undetected reduces the child's ability to extract the important information from the auditory signal. Unfortunately, the design and acoustics of the classroom leads to problems in extracting the speech signal through reduced speech intensity due to the increased distance of the student from the teacher in addition to masking from excessive background noise. One potential solution is the use of sound-field amplification which provides a uniform amplification to the teacher's voice through the use of a microphone and loudspeakers. This investigation examined the efficacy of sound-field amplification for 4 children with Down syndrome. Measures of speech perception were taken with and without the sound-field system and found that the children perceived significantly more speech in all conditions where the sound-field system was used (p < .0001). Importantly, listening performance with the sound field system was not affected by reducing the signal-to-noise ratio through increasing the level of background noise. In summary, sound-field amplification provides improved access to the speech signal for children with Down syndrome and as a consequence leads to improved classroom success. PMID- 11915434 TI - Is there a relationship between zinc and the peculiar comorbidities of Down syndrome? AB - Zinc plays a central role in the immune system and has been found to be significantly reduced in people with Down syndrome. The effectiveness of zinc supplementation in people with Down syndrome has been reported with discordant results. A comparison was made between a range of clinical and biochemical variables and zinc levels in 120 individuals with Down syndrome. Two groups of participants, one with normal zinc levels and the second with low zinc levels, were compared on the following measures: growth hormone secretion, IgA and IgG antigliadin antibodies, presence of coeliac disease, T3, T4, fT3, fT4, TSH, hypothyroidism, hyperthyroidism, CD4/CD8 ratio, total immunoglobulins G and subclasses. No significant difference was found between the two groups, except for IgG4 which was, unexpectedly, significantly decreased in the group with normal zinc levels. In conclusion, an impairment of zinc blood level in individuals with Down syndrome does not necessarily impact on the organs and systems evaluated here. PMID- 11915435 TI - Oesophageal atresia and Down syndrome. AB - Infants with Down syndrome are known to have a high frequency of associated birth defects and some authors have suggested an association between Down syndrome and oesophageal atresia. We evaluated data from the Sicilian Registry of Congenital Malformations. Our finding of an incidence of 0.9% of oesophageal atresia in children with Down syndrome is more than 30 times higher than expected and more than reported in other studies where the association was present in 0.5% of cases. Our results confirm that the relationship between Down syndrome and oesophageal atresia is a non random association and the observation of this association in several populations with different genetic backgrounds allows us to conclude that a causal relationship may exist between Down syndrome and oesophageal atresia. PMID- 11915437 TI - Individuals with Down syndrome. PMID- 11915436 TI - Cancer incidence in persons with Down syndrome in Israel. AB - The purpose of this study was to assess the incidence rates of leukaemia and other malignancies in persons with Down syndrome in Israel. The target population consisted of all persons with Down syndrome in the period of 1948-1995 and the study population was divided into two subgroups: (1) Persons born in Israel between 1979-95 (registry group) and (2) Persons currently or past institutionalised, born before 1979 (institution group). The study population was linked to the Cancer Registry and cases that had been diagnosed through December 1995 were subsequently identified. The observed incidence rates were compared to expected rates in the general population. Standardised Incidence Ratios (SIR) and 95% confidence intervals were computed for each disease category. Analyses of results were performed separately for each subgroup of our study population. In the registry group seven cancer cases were observed as compared to 1.5 expected (SIR = 4.67 95% CI 1.9-9.6), all of which were leukaemia cases. For the institution group a total of 17 cancer cases were observed compared to 12.8 expected. These included four cases of leukaemia (SIR = 6.90 95% CI 1.9-17.7). An excess of gastric cancer in males, based on two cases (SIR = 11.9 95% CI 1.3 42.9) was also observed. The significant excess of leukaemia in the Down syndrome population in Israel is in accordance with other international studies. The excess of gastric cancer in males with Down syndrome, which has not been reported before, should be further explored. PMID- 11915438 TI - Suit arising from physician assistant's services raises supervision questions. Rockefeller v. Kaiser Foundation Health Plan of Georgia. PMID- 11915439 TI - Professional association's disciplinary action upheld. Austin v. American Association of Neurological Surgeons. PMID- 11915440 TI - Proposed legislation to provide health services during air flight. PMID- 11915441 TI - [Partial purification and use of Trypanosoma cruzi glycosidic fractions for Chagas disease diagnosis]. AB - Protein and glycoprotein fractions are used for Chagas' disease diagnosis, but they are degraded by parasite endogenous proteases. Therefore protease inhibitors are used for their conservation increasing the antigen cost. The possibility of using protease-resistant glycosidic fractions could solve this problem allowing to obtain stable antigens, with a high sensitivity at a lower cost. This proposal is reinforced by the existence of anti-galactosyl antibodies against T. cruzi oligosaccharic fractions in sera from chagasic patients. The aim of this work was to obtain T. cruzi glycosidic fractions and to evaluate their possible use for Chagas' disease serologic diagnosis. Total protein and glycoproteins from the four stages of T. cruzi were obtained. The glycosidic fractions were obtained by exhaustive proteolysis with Proteinase K. Protein, glycoprotein and glycosidic profiles were analysed by SDS-PAGE followed by staining proteins (Coomassie/silver) or glycoproteins and glycosidic fractions (APABGP). Antigenicity of the different fractions was determined by Western Blot and luminography, using control hyperimmune serum anti-epimastigote and a "pool" of chagasic human sera. Finally, the capacity of the glycosidic fractions to discriminate chagasic and non-chagasic human sera and the sensitivity of the fractions respect to control serum, were determined by ELISA. Main findings were: a) there are peptides, glycopeptides and glycosidic fractions that are common and specific to parasite stages; b) there are more antigenic glycoproteins in epimastigotes and metacyclics than in trypomastigotes and amastigotes; c) there is a correlation between the glycosidic fraction-pattern and the host type; d) glycosidic fractions can be used as antigens to discriminate, by ELISA, chagasic and non chagasic patients, but show lower titers with respect to total proteic and glycoprotein antigens. It is concluded that it is possible to develop a diagnostic kit for Chagas' disease employing glycosidic fractions, eliminating the disadvantages of the current kits. PMID- 11915442 TI - [Leaf anatomic studies in solanaceae of Venezuela. VI. Leaf anatomy of 10 species of Cestrum L]. AB - The leaf anatomy of Cestrum acuminatissimum Dunal, C. alternifolium (Jacq.) O.E. Schulz, C. glabrescens (C.V. Morton) Steyerm. et Maguire, C. imbricatum Rusby, C. Iatifolium Lam., C. neblinense D'Arcy et Benitez, C. ruizteranianum Benitez et D'Arcy, C. schulzianum Francey, C. tillettii Benitez et D'Arcy y C. tubulosum Sendtner, is described in order to value its diagnostic characters useful as source of data in the taxonomy of the Solanaceae. The material was obtained from specimens deposited in MY herbarium, coming from different geographical areas of Venezuela, and it was prepared according the classical methodology used in leaf anatomy studies. The results obtained have, besides the common features mentions for other species of the genus Cestrum, some differences in: types and density of trichomes, thickness and sinuousities of the cell wall, stomata position, as well as in the distribution of esclerenchymatous cells of the mesophyll, midvein petiole. PMID- 11915443 TI - [Abortion level in 5 species of Cestrum L. (Solanaceae)]. AB - We studied the fitness and abortion levels in five species of genus Cestrum L. (Solanaceae) present in a cloud forest of Parque Nacional Henri Pittier, Venezuela. The seed set is variable and the abortion is higher at flowers-fruits and ovule levels, while the S/O ratio is low. We discuss the possible causes of abortion and seed set in these species. PMID- 11915445 TI - [Influence of age and gender on plasma norepinephrine changes produced by orthostatic stress]. AB - The aim of this work was to determine the effect of aging and gender on plasma norepinephrine (NE) response to orthostatic stress. Fifty-six men and sixty women non-smokers, sedentary, non-obese, were studied at supine and standing positions. On the basis of the age of the subjects, they were divided into three groups of either men or women. Group A, young, range 17-34 years, Group B, middle-age, range 40-60 years, and Group C: senescent, range 61-91 years. Senescent subjects had the highest absolute supine values of NE and significant differences between women and men were found in groups B Men: 298 +/- 15 pg/mL; Women: 348 +/- 14 pg/mL), and C (M: 386 +/- 29 pg/mL; W: 473 +/- 19 pg/mL), but not between young (M: 246 +/- 21 pg/mL; W: 261 +/- 18 pg/mL). NE increased markedly in response to the orthostatic stress but the relative (percentage) increases were smaller in the older subjects perhaps related to their elevated baseline levels. The highest percent positive variation was observed in group A (M: 85%; W: 82%). The oldest group (C), had the lowest variation (M: 23%; W: 21%). The relative increases in NE responses were not different in men and women. The orthostatic-induced variation in NE was negatively related to aging in both men and women (M: r = 0.654; p < 0.0001; W: r = -0.557; p < 0.0001). It is concluded that orthostatic stress induced rise of blood NE is attenuated by age but does not appear to be dependent on gender. PMID- 11915444 TI - [Inventory of reptiles in 2 semi-arid zones from Northeastern of the Peninsula de Araya, Sucre State, Venezuela]. AB - The fauna of reptiles in two localities from Northeastern Peninsula de Araya (Guayacan and El Morahal), Sucre State, Venezuela, was evaluated. Both zones are characterized by a vegetation of thorny tropical mount type, and semiarid climate of scarce precipitations (less than 700 mm). Field trips were made between june 1997 and june 1998. The samples were collected both during day and night, with the aid of conventional accessories. The information was complemented with visual registrations and bibliography revision. A total of 21 species were captured and/or observed, distributed in 10 families belonging to 2 of the 3 orders present in Venezuela. The most important families from the point of view of the diversity of species, were the Gekkonidae (2.00 bits/species) for the lizards and the Colubridae (2.33 bits/species) among the snakes. It was also reported one species of tortoises and three of cinegetic interest, being Cnemidophorus lemniscatus, Ameiva bifrontata y Tropidurus hispidus the only species of constant presence during the study. PMID- 11915446 TI - [Nutritional evaluation of green plantain flour dehydrated soups. Starch in vitro digestibility]. AB - Previous works have shown that green plantain flour (GPF) contains a considerable amount of resistant with similar effects to dietary fiber. In order to diversify the use of this fruit the purpose of present study was to formulate and elaborate powered, dehydrated, cream type soups with green plantain flour flavored with vegetables (onion, coriander and leak) which increase the dietary fiber content of the preparation. Green plantain was peeled, cut in medium size pieces and submerged in 0.1% citric acid solution. The dehydration process was forced air drying (80 degrees C), followed by milling. The same procedure was applied to the flavoring vegetables. To obtain the cream type soups various formulations were tried containing 50-63% resistant starch, 11.7-12% dietary fiber 6.5-6.9% protein. The mineral content of the preparations is reported. Viscosity of 1:10 (w/v) soups was 630-670 cps. In vitro starch digestibility after 6 hours was 38% with porcine amylase, increasing to 48% if the enzyme was from bacterial origin, supporting previous results that suggest resistance to hydrolysis of green plantain (GP) starch granules. In conclusion this study diversifies the use of GP and suggests that dehydrated GPF soups due to their high dietary fiber, resistant starch content and to the slow starch hydrolysis may be used in special nutrition regimes. PMID- 11915447 TI - [New observations on the structure of the lingual filiform papillae epithelium of the toad Bufo marinus L.: Light and transmission electron microscopy study]. AB - There are several inconsistencies and confusions about the microscopic structure in the anurans dorsal, non-sensory, mucosal epithelium of the tongue. The aim of this work was to differentiate the structure of the epithelium covering filiform papillae from the glandular epithelium. For this, samples of the dorsal distal mucosa of Bufo marinus tongue were processed for light microscopy (LM) and transmission electron microscopy (TEM). For LM, stained 1 micron sections were used. The results showed that the filiform papillae were covered by a biestratified epithelium formed by a basal layer of dendritic cells and an upper surface layer consisting of numerous seromucous secretory cells, dendritic cells apical processes, and some goblet cell-like and ciliated cells. LM revealed that most filiform papillae showed the seromucous secretory cells in apparent resting state. These cells were characterized by a basal, round nucleus, pale cytoplasm with small, dispersed, stained granules in the supranuclear region. With TEM, seromucous secretory cells exhibited poorly developed RER, scattered medium electron dense secretory granules and microridges at the apical surface. Dendritic cells were seen as branched cells completely surrounding the basolateral surfaces of secretory cells. In addition, some dendritic cell processes have microvilli at their abutting endings. Goblet cell-like cells theca showed a continuous mesh of medium electron dense, partially fused, mucous granules and irregular microvilli-like processes at the apical end. Ciliated cells were mostly localized at the lateral wall of filiform papillae. Results suggest that at the filiform papillae, yuxtacrine or paracrine interactions mediated by physical contacts between dendritic and secretory cells may exist. PMID- 11915448 TI - [The phorophyte occupation patterns by vascular epiphyte species in the High Orinoco tropical humid forest, Amazonas State, Venezuela]. AB - By international agreement (Austria-Venezuela), an experimental area was selected for canopy ecology studies. A tower crane was installed that allows direct access to the canopy of 1.5 ha area of the forest. This is a quantitative study of the phorophyte occupation patterns by vascular epiphyte species in an Amazon tropical humid forest. The forest of Surumoni consists roughly of four arboreal strata, i.e., emergent (conspicuous), superior, middle and lower. The largest proportion of trees occurs in the lower diametric classes (smaller DBH), and the total distribution approaches an "inverted J" curve. The most abundant tree species in the three strata is Goupia glabra (Celastraceae); making this forest a case where a single arboreal species dominants. Mean of epiphyte individuals per occupied phorophyte is too variable, depending on phorophyte species and showing no correlation with occupied tree percentage. The species Eschweleira parviflora (Lecythidaceae) represented in the study area by a single individual of the superior stratum with high density of vascular epiphytes, indicates that this support species is particularly favorable for the establishment epiphytic flora. PMID- 11915449 TI - [Molecular and cell aspects of the cancer metastasis]. AB - The metastasis dissemination is a process which leads to a primary tumor cells to migrate, infiltrate host tissues and form a secondary tumor focus at distance. This clinic evolution is a consequence of the cancer natural history and is due to the appearance of new potentialities in tumoral cells, providing to a small quantity of them an invasive and metastatic capability. This dissemination is possible by many factors; among them the lack of cohesion in tumor tissue cells, forming new blood vessels, resistance to anoikis and posterior implantation of tumoral cells in a heterotypic site. The change of cohesion in cell systems, synthesis of proteolytic enzymes, cell motility, modification of signals induced by growth factors, or the substratum for environment adhesion, and lack immune recognition by the immunologic system, are the main contributors to the appearance of metastasis. A permissive ecosystem is necessary to implant tumoral cells in target organs such as: liver, lung and bone marrow. This special environment, which is more favorable to metastasis, supplies an stroma, adhesive systems, growth factors and neo-vascularization. It is important to point out that the development of a metastasis is preceded by some genetic changes which make possible the adaptation of malign cells to a new microenvironment. PMID- 11915450 TI - [Implants in the posterior maxilla. A case report]. PMID- 11915451 TI - [Clinical experiences with adhesive fixed, glass fiber reinforced inlay bridges. Evaluation of a prospective clinical study after 24 months]. PMID- 11915452 TI - Current state of cement fixation in THR. AB - The author surveys important landmarks in the development of total hip arthroplasty, with an accent on implant fixation using acrylic cement. He explains why he personally opted for hybrid prostheses, combining a cemented stem and a cementless socket, in patients over sixty years. Excellent cementless, sockets have been available for a long time; on the femoral side, the first steps were difficult, but several cementless, stems were subsequently developed, which provided excellent long term results. This historical evolution resulted in a very uneven use of cemented versus cementless stems from one country to another in Europe. Cemented implants have enjoyed a renewed popularity over the past few years as a result of several factors, including economical factors. The author discusses the conditions for optimal fixation of a cemented stem; these conditions are not always met satisfactorily, as a number of surgeons obviously stick to a crude cementing technique. The author describes the role of the stem geometry and surface finish, as well as the possible influence of a centralizer; he explains why, based on a correct analysis of the available data, discredit has been unduly cast on cemented stems made of titanium alloy. He insists on one important although often disregarded factor: the specific type of cement used, as better results have clearly been achieved with certain cements than with others. He insists on the necessity to take into account all the elements involved, in order to avoid making erroneous conclusions. He also insists on one very important variable, the quality of the surgical technique. Total hip arthroplasty is likely to make further progress in the future, although we are likely now in the asymptotic portion of an ascending curve. Further improvement in clinical results will result from improvement of currently existing systems and optimization of surgical technique, rather than from the continuous designing of new implants. PMID- 11915453 TI - Delayed post-traumatic osteonecrosis of a vertebral body (Kummell's disease). AB - Kummell's disease is a rare spinal disorder characterized as avascular necrosis of a vertebral body occurring in a delayed fashion after minor trauma. The disease derives its name from the German surgeon Hermann Kummell, who described a series of patients with the disorder in 1895. Kummell's disease is distinguished from typical osteoporotic compression fractures, by virtue of the fact that patients with Kummell's disease develop symptoms in delayed fashion. In addition, Kummell's disease exhibits different signal characteristics on magnetic resonance imaging when compared to osteoporotic compression fractures. In this review we discuss the pathophysiology, histopathology and treatment of this interesting disorder. PMID- 11915454 TI - Ligament reconstruction/tendon interposition arthroplasty for thumb basal joint osteoarthritis preliminary results of a prospective outcome study. AB - Ligament reconstruction/tendon interposition arthroplasty (LRTIA) is a common procedure for basal joint osteoarthritis of the thumb. We evaluated 31 operations in 30 patients, 28 females, 2 males with a mean age of 57 years. The mean follow up was 24 months (range 8 to 46 months). Pain reduction was 75% on average, with good preservation of motion and of first web space. Twenty-six patients were satisfied or very satisfied; five were not. The grip strength and key-pinch strength increased post-operatively, albeit not significantly. PMID- 11915455 TI - Hydroxyapatite-coated stems with metaphyseal and diaphyseal press-fit. Eleven year follow-up results. AB - We report the 11-year follow-up results of 52 unilateral primary hip arthroplasties performed with hydroxyapatite-coated stems. The femoral prosthesis used was a collarless titanium alloy implant, with proximal circumferential hydroxyapatite coating and increased distal thickness to fit the proximal diaphyseal region of the femur. Clinical evaluation was performed using the Merle d'Aubigne Hip Score. Anteroposterior and lateral radiographs were obtained and compared with previous postoperative films. Radiographic evaluation was carried out following Engh's criteria for uncemented implant fixation and using Livermore's method for measurement of polyethylene wear. At the end of the follow up period, excellent and good clinical results were recorded in 40 arthroplasties (77%). The incidence of thigh pain at one year was 32.7%, but it decreased to 4.2% after the first post-operative year. The 11-year survival rate was 92.3%. Seven arthroplasties were revised because of aseptic loosening of the cup in one case, aseptic loosening of the stem, in one case, septic loosening of the stem in one case, periprosthetic fracture in two cases and polyethylene wear in three cases. Forty-two (87.5%) of the nonrevised stems met the criteria for radiographic osseointegration. Cortical hypertrophy was observed around the mid part and tip of the stem in 22 patients of the series. This sign tends to be related to thigh pain (p < 0.1). Calcar osteolysis was present in 8 cases. There was only one case of distal femoral osteolysis. We found a strong and significant relationship between long-term wear rates and the occurrence of osteolysis (p < 0.001). We concluded that thigh pain is in relation to the distal diameter of the stems and significantly decreases after the first postoperative year. There was a low incidence of osteolysis in our series in comparison with other series of noncemented implants with 32-mm femoral heads and with similar follow-up. PMID- 11915456 TI - The effect of femoral stem size on failure rates in total hip replacement cemented with Boneloc. AB - We report the clinical and radiological outcome of 111 primary Taperloc hip arthroplasties implanted with Boneloc bone cement and the effect of stem size on survival. The mean follow-up was 5.3 years (range 3.2-6.6 years). The average age of the patients at operation was 73.4 years. Twenty-seven were male and 84 female. We defined clinical failures as those who had revision for aseptic loosening, or those symptomatic with pain and subsequent confirmation of radiographic loosening. The overall failure rate noted for this type of stem was 20.7%. Failure occurred on average at 3.3 years (0.3-6.3 years). However, when the data is broken down to different sizes of stem, the patients who had smaller Taperloc stems (7.5 mm or 10 mm) had a 27% failure rate whereas stems equal to, or greater than, 12.5 mm had a 12% failure rate over the same period (p < 0.05). As compared to other types of femoral stems used with Boneloc cement, the Taperloc stems survive better. Furthermore, larger stems survive even better. We suggest that these larger stems behave more like uncemented femoral stems. The finish on the stem is not a major contributor as has been suggested in the past. PMID- 11915457 TI - Pelvifemoral external fixation for the treatment of open fractures of the proximal femur caused by firearms. AB - Seventeen patients with open fractures of the upper third of the femur were treated using a pelvifemoral external fixation device. All of them had grade III open fractures resulting from high-velocity missile and explosive injuries with massive foreign body contamination. Sciatic nerve injury was present in five (29.4%); abdominal viscera and thoracic wall injuries were present in two patients (11.8%). There were no major arterial injuries. Full weight bearing was allowed after clinical and radiological bone healing (average 11.5 months). Chronic osteitis with fistula and sequestra developed in two (11.8%) patients. There were no nonunions and no refractures. Minor painless limitation of hip motion persisted in all patients. Upper-third femoral open fractures due to firearms are a unique type of open fractures. They are usually highly comminuted; therefore, stable fixation is difficult or impossible to achieve using external fixation with transfixation of the fracture site. On the other hand, the risk of infection is high following intramedullary nailing. Pelvifemoral external fixation allows adequate management of the soft tissue wounds, provides stable bone fixation and allows early patient mobilization. PMID- 11915459 TI - The reliability of the pre-operative classification of open tibial fractures in children a proposal for a new classification. AB - The purpose of this observational study was to evaluate the accuracy of Gustilo's classification of open tibial fractures in children. Pre- and post-debridement (using the parameters of Gustilo's classification) wound gradings in 27 children with a mean age of 10 years (3 to 15 years) who had sustained an open tibial fracture were compared. Pre-operative Polaroid photographs of the wound were taken of all these patients in the accident and emergency department. In every case, the fracture was treated with prophylactic intravenous antibiotic administration, wound debridement and lavage. Following wound exploration in the operating theater, the wound was classified using Gustilo's parameters again; this was different from the initial grading. We compared post-debridement classification according to Gustilo to the new classification which we propose. The latter classification is a peroperative assessment of the extent of soft tissue damage and it addresses bone stability. According to this classification, the majority of open tibial fractures were stable, requiring no skin graft or flap and had a good outcome. Only five patients were treated by initial external fixation of the tibia; the remainder were treated by cast immobilization. Wounds were treated as appropriate. The clinical outcome study included the assessment of wound and fracture healing and the incidence of complications. The mean period for follow-up was 8 months (6 to 24 months). There were no cases of nonunion or deep wound infection and the wounds healed in all these patients. We conclude that Gustilo's classification is not specific and does not reflect the extent of soft tissue and skeletal damage. Factors such as the degree of soft tissue damage and periosteal stripping that are noticed following wound debridement and velocity of injury are far more important than the wound size. Our proposed peroperative classification covers the extent of soft tissue injury and skeletal stability, thus predicting the outcome more than the Gustilo classification. PMID- 11915458 TI - Effect of dynamization in open interlocking nailing of femoral fractures. A prospective randomized comparative study of 50 cases with a 2-year follow-up. AB - This prospective randomized comparative study evaluated the role of dynamization of interlocking nails after open reduction and internal fixation of femoral shaft fractures. Fifty femoral shaft fractures were treated by open interlocking nailing and were statically locked. Twenty six of these 50 patients were randomly selected for dynamization and the other 24 were treated without dynamization. The patients were followed up for at least 2 years. The 26 cases which were dynamized went on to union between 13 and 28 weeks (average 19.2 weeks) with two poor results, including one nonunion. The cases that were not dynamized went on to union between 16 and 30 weeks (average 23.5 weeks) with two poor results, including one nail breakage. Though final results are comparable, the study suggests that dynamization after open interlocking nailing significantly shortens the mean time to union, though it does not significantly affect the union rate of the femoral shaft fractures. PMID- 11915461 TI - Modified shoelace technique for delayed primary closure of the thigh after acute compartment syndrome. AB - The authors report a case of acute compartment syndrome in the thigh in a 19-year old man with multiple injuries including fracture of the femoral diaphysis. Decompressive fasciotomy was performed emergently. Complete progressive closure of the wound without split-thickness skin grafting was achieved using a modified shoelace technique: sutures were run inside wide drains placed in contact with the muscles and were then tightened over the skin. These drains enlarged the contact area between sutures and muscles, preventing damage to muscles. PMID- 11915460 TI - [Triple arthrodesis in the management of acquired flatfoot deformity in the adult secondary to posterior tibial tendon dysfunction. A retrospective study of 20 cases]. AB - The authors have reviewed 20 cases of valgus flatfoot deformity in the adult, secondary to insufficiency of the tibialis posterior tendon, which were treated by triple arthrodesis from 1983 to 1998. The longitudinal arch was flat grade 3 in 18 cases and grade 2 in two. The rearfoot was in valgus in all cases. Dorsal flexion of the ankle was limited to 5 degrees with a short Achilles tendon in 10 cases Djian-Annonier's angle was 134 degrees on average. The talocalcaneal divergence was 32 degrees. Diffuse osteoarthritis was present preoperatively in 14 cases. Fusion of the arthrodesis was achieved in 18 cases. There were two nonunions: one affecting the talonavicular joint, and the other, the talonavicular and calcaneocuboid joint. Results evaluated according to Kitaoka's criteria were excellent in 35%, good in 35%, fair in 20% and poor in 10%. Xrays showed progression of osteoarthritis. These results were not as good as in Fortin and Waling's series but we did not use iliac grafts. There was no instance of talus necrosis. The nonunion rate was higher than in other series at the talonavicular level. In order to improve the results, we now believe that bone resection must be kept minimal and bone grafts should be used, in addition to internal fixation. Triple arthrodesis was found to give good result in the treatment of acquired flat foot deformity due to insufficiency of the tibialis posterior tendon. However on the long term it promotes degenerative changes in neighbouring joints. PMID- 11915462 TI - Sweet's syndrome associated with pigmented villonodular synovitis. AB - Sweet's syndrome (acute febrile neutrophilic dermatosis) was first described in 1964. The typical symptoms of Sweet's syndrome are high temperature, peripheral leucocytosis, painful cutaneous rashes (papules, plaques) and arthralgia. Sweet's syndrome has particularly been described in association with neoplastic, infectious and immunological diseases. The pathogenesis of Sweet's syndrome can be explained by a reaction to an antigenic structure with accumulation of immunological complexes and liberation of inflammatory mediators. For the first time we report on a patient with Sweet's syndrome and pigmented villonodular synovitis, which is believed to play the antigenic role in the Sweet syndrome. PMID- 11915463 TI - Supracondylar process of the humerus. AB - Four cases of supracondylar process of the humerus in three patients are presented. The main features of a supracondylar process as compared with an osteochondroma are reviewed. The three patients had pain and one had signs indicating median nerve compression. One had a supracondylar process together with an osteochondroma in the contralateral supracondylar region. One patient with a bilateral supracondylar process refused an operation. The other two patients underwent surgical treatment. The symptoms disappeared in the two patients who were operated. PMID- 11915464 TI - Angioleiomyoma of the knee. A case report. AB - The authors report a case of an angioleiomyoma at the anteromedial side of the knee mimicking a neurinoma of the infrapatellar branch of the saphenous nerve. Angioleiomyomas are benign soft tissue tumors with a predilection for the lower extremities in middle-aged females. Paroxysmal pain is triggered by pressure, cold and hormonal changes. Simple excision is usually curative. PMID- 11915465 TI - Abdominal hernia through a defect in the iliac bone after resection of a chondrosarcoma of the pelvis. AB - Chondrosarcoma is a malignant tumor with, generally, a better prognosis than osteogenic sarcoma; it usually calls for surgical treatment with a wide margin of resection, in view of its marked resistance to chemo- and radiotherapy, but this may be difficult or impossible in one of its most frequent locations, i.e. the pelvis. Herniation of abdominal contents through a surgical defect in the iliac bone is a rare complication, and is almost always associated with bone graft harvesting from the iliac crest. We present the case of a male patient operated for a chondrosarcoma of the ilium, with wide resection of the tumor. During follow-up in the outpatient department, the existence of a hernia with intestinal contents through the defect created in the ilium was noted. The patient has now been followed for 10 years, and there have been no clinical repercussions of the hernia. No recurrence of the tumoral lesion has been observed. We review the cases published and discuss the treatments recommended. PMID- 11915466 TI - Dislocation of the elbow with intra-articular entrapment of the medial epicondyle in adults. Report of two cases. AB - Intra-articular entrapment of the medial epicondyle with dislocation of the elbow is a common injury pattern in childhood. The occurrence of this injury is extremely rare in adults. We present two cases of dislocation of the elbow with intra-articular inclusion of the medial epicondyle in adult patients. One case was treated by closed reduction and the other required open reduction. A brief review of the scanty literature on the subject is also presented. The case reports highlight the need for early diagnosis and management of these atypical cases in adults. PMID- 11915468 TI - [Surgery for craniosynostosis: timing and technique]. AB - Craniosynostoses are a group of diseases, the presentation of which differs markedly on account of the cranial suture involved. Their impact is cosmetic, cerebral, and ophthalmologic. Syndromic craniosynostoses associate a more or less pronounced faciostenosis, which requires surgical correction as well, because of cosmetic, ophthalmologic or airway problems. Surgical treatment depends on the type of the craniosynostosis and the patient's age; ideally, the child should be operated between 3 and 12 months. This surgery requires a perfect collaboration between neurosurgeon, plastic surgeon, and anaesthesiologist. Surgical correction allows in large measures the preservation of intellect, sight, and body image. PMID- 11915467 TI - [Multicentric giant cell tumor of the upper arm]. AB - Multicentric giant cell tumors of bone are rare; they represent less than 1% of all giant cell tumors. We report the case of a 35-year-old right-handed man, who presented in 1985 with a giant cell tumor of the upper end of the right humerus. After failure of conservative treatment (curettage and bone grafting), resection arthrodesis of the shoulder with a free vascularized fibular autograft was performed. Three years later, the patient developed an osteolytic lesion of the lower end of the ipsilateral radius, involving the soft tissues and the wrist joint. He was treated with resection-arthrodesis of the wrist with a free vascularized fibular graft. The histologic study confirmed the diagnosis of giant cell tumor of similar grade. After 13 years follow-up, the patient had a satisfactory function of the limb and no recurrence of the tumor was noted. PMID- 11915469 TI - [Anesthetic management for craniosynostosis]. AB - Surgical procedures for correction of craniosynostosis are performed in young infants with a small blood volume and represent major surgery with extensive blood loss. An accurate determination and a precise restoration of blood losses represent the major concern for the anaesthetist during this surgery. The preoperative assessment of these patients is usually simple, except in the cases where the craniosynostosis is associated with other congenital malformations. The anaesthetist should keep in mind that intracranial hypertension may be associated with craniosysnostosis, which modify the anaesthetic management, especially the induction of anaesthesia. Even though the psychological impact of a craniosynostosis should be taken into consideration, surgery is most often indicated for functional considerations, therefore parents should be informed of the risks related to the procedure. During the postoperative period the major concerns are related to the possibility of a persistent bleeding, which usually decreases and disappears over the first 12 hours. PMID- 11915470 TI - [Perioperative management of blood loss during surgical treatment for craniosynostosis]. AB - Blood saving is the major challenge during the surgical repair of craniofacial deformities. Treated patients have a low reserve volume and the techniques available to lower homologous blood transfusions are limited or insufficiently evaluated in this particular case. The most important factor determining blood loss is the quality of the surgical haemostasis. Blood saving begins with early preoperative evaluation of the patient's bleeding risk, which is a function of the type of surgery, of the surgical technique, of the number of sutures involved, of the length of surgery, and of the patients age, weight and physical status. Elaborated blood saving techniques such as preoperative autologous blood donation, erythropoietin administration, normovolaemic haemodilution, and peroperative autologous blood saving and reinfusion have revealed disappointing where used alone. These techniques require a heavy setup and still need to be evaluated extensively. They should be used in selected cases such as in patients with a very high risk of bleeding or face to Jehovah Witnesses. Monitoring during surgery should include precise evaluation of blood losses and haematocrit measurements at regular intervals. The haematocrit threshold allowing homologous blood transfusion should be set at 21%, provided that any other source of autologous blood is exhausted. Postoperative monitoring should also include precise evaluation of blood losses and haematocrit measurements. The 21% threshold should remain the reference during that period. PMID- 11915471 TI - [Postoperative analgesia in pediatric neurosurgery]. AB - Pain management in paediatric neurosurgery must be a daily concern for surgeons and anaesthetists. Pain assessment in infants and small children is difficult to perform because of limitations associated with these patients. The association of safe and effective analgesics allows good pain relief together with good safety conditions. However, neuropathic pain, which may occur following neurosurgical procedures, will require further studies. PMID- 11915472 TI - [Traumatic head injury in children: physiopathology and clinical management]. AB - Traumatic brain injury (TBI) constitutes a major health and economic problem for developed countries, being one of the main causes of mortality and morbidity in children. In a busy traumatology center, a child will be admitted daily in the emergency department with head trauma injury. The anaesthesiologist must have a complete understanding of the pathophysiology and develop a practical knowledge of initial management of such patients. Traumatic brain injury may have intracranial and systemic effects that combine to give global cerebral ischaemia. Injury to the nervous system, irrespective of the primary injury, initiates a multitude of inflammatory cascades resulting in secondary brain injury. The consequence of these secondary brain injuries is most often as important, if not, more important than the primary injury. This period of brain inflammation can last up to three weeks and renders the brain more susceptible to the effects of systemic insults such as hypotension, hypoxia and or pyrexia. It has been shown in post-mortem examination of patients dying from severe traumatic brain injury that more than 91% had evidence of secondary ischaemic damage. These secondary injuries may be responsible for the clinical presentation of the "child who talk and die". The concept of "cerebral protection" has been extended to encompass the active treatment of secondary injury and the prevention of cerebral ischaemia. Initial care focuses on achieving oxygenation, airway control and treatment of arterial hypotension. PMID- 11915473 TI - [Severe head injuries in the young child: early management]. AB - The initial management of severely head-injured patients, including infants and children, is aimed at preventing and treating secondary brain damage, which mainly result from systemic insults (hypoxaemia, hypercarbia, arterial hypotension). Orotracheal intubation, followed by continuous sedation-analgesia, is mandatory when the Glasgow Coma Scale score (GCS) is less than or equal to 8 (crush induction is recommended). The goal of mechanical ventilation is to maintain normoxaemia and normocarbia. Moreover, the maintenance of an optimal cerebral perfusion pressure, usually 50 mmHg in infants, requires volume loading (isotonic fluids and colloids), and catecholamines if arterial hypotension persists. Intravenous mannitol is used only in case of life threatening intracranial hypertension, keeping in mind the potential for aggravating an hypovolaemia. Cerebral tomodensitometry is the most relevant imaging procedure for diagnosing surgical brain lesion. However, it should be noted, that severe head trauma is frequently associated with extra-cranial traumatic injuries, which may be responsible for (avoidable) deaths if the diagnosis is not made or delayed. Therefore, infants and small children presenting with severe head trauma should be considered as multiple injured and treated accordingly. Adequate initial management of severely head-injured children may participate to improved neurological outcome. PMID- 11915474 TI - [Multimodal monitoring of head injuries in children]. AB - As in the case of adults, there are three main goals in the monitoring of severe head trauma in children: to prevent or minimize the apparition of secondary lesions, to optimize treatment, to help make precise prognosis. The basic monitoring is composed of repeated clinical examinations, brain radiological imaging and control of vital parameters (blood pressure, temperature, PaO2 (SpO2), PaCO2 (FETCO2), haemoglobin, haematocrit. On the other hand, during specific brain monitoring, the brain perfusion (TCD, intracranial pressure), the electrical activity of the brain and sometimes the brain oxygenation (SvjO2) are controlled. The data obtained from the brain monitoring must always be interpreted carefully. A child with a severe head trauma, in ICU, always requires constant and competent medical attention. PMID- 11915475 TI - [Follow-up study of head-injured children]. AB - The authors reviewed in a retrospective study 61 cases of severely head injury children who where admitted in the rehabilitation centre to evaluate the outcome 1 and 5 years after the brain injury. Neurologic and neuropsychologic status of children was assessed ad admission, 1 and 5 years later. Duration on intubation and age at time of head trauma were the worst functional prognosis. PMID- 11915476 TI - [Clinical applications of stereotaxic methology]. AB - Cerebral stereotaxy is an old methodology allowing an accurate approach of a lesion or a function, in constant renewal with the introduction of computers and robotic. There is a natural complementarity with recent neuroradiological investigations and together, it is possible to reach cerebral deep-seated or functional structures with inocuity and fiability for diagnosis and/or therapy. Its application is very large and also influences neuronavigation procedures, current in conventional neurosurgery. Tumoral stereotaxy is commonly used and achieves a better adaptation of the therapeutical strategy according to the lesions' site and histological diagnosis. The development of functional stereotaxy is associated with the interest of the neurosurgical treatment of involuntary abnormal movements, without forgetting different aspects of surgery of chronic pain and intractable epilepsies. Moreover, the stereotactic methodology leads the concept of radiosurgery, which is in some indications a true alternative to open surgery (arteriovenous malformations, vestibular schwannoma, metastasis) under the control of accurate selection in a multidisciplinary approach. PMID- 11915477 TI - [Anesthetic particularities of stereotaxic neurosurgery]. AB - Functional neurosurgery procedures are long and specific. Cooperation of the patient may be necessary during surgery. The interference of anaesthetic agents with electrophysiological monitoring should be as little as possible. Local anaesthesia combined with intravenous sedation is often used, but general anaesthesia is more comfortable and secure. Since awakening during the procedure is generally planed, it has to be quick, reliable and of excellent quality. These requirements are fulfilled by the association of propofol by target-controlled infusion (TCI) and a continuous infusion of remifentanil. PMID- 11915478 TI - [Pediatric neuroanesthesia]. PMID- 11915479 TI - [The specificity of neurosurgical anesthesia for the child]. AB - Anaesthesia for paediatric neurosurgical procedures presents an interesting challenge to the anaesthesiologist. The child is not simply a small adult. At birth the central nervous system (CNS) development is incomplete and will not be mature until the end of the first year of life. Because of this delay in the maturation of the CNS, several specific pathophysiological and psychological differences ensue. Although one has little control on the child primary lesion, the selection of an anaesthetic technique designed to protect the perilesional area and the recognition of perioperative events and changes may well have a profound effect in the reduction or prevention of significant morbidity. Current neuroanaesthestic practice is based on the understanding of cerebral anatomy and physiology. Paediatric neuroanaesthesiologists must face the added challenge of the physiological differences between developing children and their adult counterparts. PMID- 11915480 TI - [Cerebrospinal fluid shunting: anesthetic particularities]. AB - The symptomatic treatment of hydrocephalus remains cerebrospinal fluid (CSF) drainage to an external reservoir (external CSF drainage) or to an internal cavity mainly the peritoneum or the right atrium via a unidirectional valve (internal CSF drainage) and finally by endoscopic ventriculocisternostomy. Local anaesthesia is adequate for external CSF drainage in adults and children above 10 years while general anaesthesia is required in all other cases. The main problems encountered in these patients are difficult intubation and full stomach associated with increased intracranial pressure. The anaesthetic approach should favour homeostasis. With the exception of ketamine and enflurane, the majority of anaesthetic drugs can be used. Anti-epileptic drug are mandatory. Antibioprophylaxis mainly against staphylococcus is systematic in internal CSF drainage. Rapid emergence from anaesthesia and extubation should be encouraged. Complications (infectious, mechanical and bleeding kinds) are frequent and are often the cause of reinterventions or revisions of the device, exposing the patients to iterative anaesthesia. Furthermore, patients with shunts are at risk of malfunction of the device when exposed to situations like pregnancy, magnetic resonance imaging, or laparoscopy. Under these circumstances, it is recommended to associate the neurosurgical team in the management of these patients and to verify that the shunt is working well before and after the procedure or event. PMID- 11915481 TI - [Infections of cerebrospinal fluid shunts in the child: prevention and treatment]. AB - Cerebrospinal fluid (CSF) shunting has developed into the mean method of treatment in children with hydrocephalus. Until the last decade, shunt infection was the most important cause of morbidity with a mean rate of 10%. Most of shunt infection (> 90%) are diagnosed within six months after surgery supporting a basic premise of direct contamination at the time of surgery. However, after applying stricter operative and perioperative protocols, some authors reported a dramatically decrease in the incidence of infectious complications. The overall annual risk of shunt infection in a paediatric neurosurgical unit is currently 1%. Risk factors are analysed with emphasis on the choice of preventive treatment. An outline of the protocol for shunt implantation is presented. There is no clearly defined role for prophylactic antibiotic medications in the prevention of shunt infection. The small sample sizes of prospective controlled clinical trials precluded sufficient statistical power. The conclusions of the meta-analyses are not sufficiently robust to resolve the controversy and it is not possible to make recommendations either for or against the use of prophylaxis in shunt surgery. The management of shunt infection is examined with emphasis on antibiotic therapy. PMID- 11915482 TI - [Anesthesia-resuscitation for intracranial expansive processes in children]. AB - The most frequent space-occupying cerebral lesions in children are brain tumors, mostly posterior fossa tumors and haematoma resulting from arteriovenous malformation rupture. They result in intracranial hypertension, directly or by compression of the cerebrospinal fluid pathway resulting in hydrocephalus. Their localization and compressive effects are responsible for specific neurological deficits and general problems. Posterior fossa lesions carry a high risk of obstructive hydrocephalus, cranial nerves palsy and brain stem compression, pituitary and chiasmatic tumors a risk of blindness, pituitary deficiency and diabetes insipidus, and cortical tumors a risk of motor deficit and epilepsy. All these parameters must be analyzed before choosing anaesthetic protocols, and surgical techniques. In the presence of life-threatening intracranial hypertension, emergency anaesthetic induction, tracheal intubation and ventilation are life-saving. The specific treatment consists in either hydrocephalus derivation, initial medical treatment with osmotherapy, or rarely surgical removal. In other situations, surgical process requires a highly deep, stable anaesthesia with perfect control of cerebral haemodynamics. Surgical positioning is complex for these long lasting procedures and carries specific risks. The most common is venous air embolism in the sitting position that must be prevented by the use of specific measures. In the postoperative period, the risk of neurological and general complications commands close surveillance, fast track extubation must be adapted on an individual basis. PMID- 11915483 TI - [Adult cranial injury in physical therapy: awakening from coma (short text)]. PMID- 11915484 TI - [Pseudoperiodic and paroxysmal electroencephalographic activities]. AB - Periodic and paroxysmal EEG patterns are frequent and easily recognized. They often correlate with particular neurological or systemic conditions. We propose a reclassification and critical examination of these periodic EEG patterns by reviewing the original work presented by J. Gaches in this journal some thirty years ago. Periodic EEG activity is still classified by its localisation- generalised or focal--and by its periodicity--long or short. Periodic long interval diffuse paroxysmal patterns are reported with rare pathologies such as SSPE, trypanosomiasis or intoxication with phencyclidine. Suppression bursts (SB) may be found in two main situations post-anoxic encephalopathies and drug-induced comas. Ohtahara syndrome is a very rare childhood epileptic syndrome with SB. Periodic short-interval diffuse pattern have been reported in Creutzfeldt-Jakob disease but also in toxic encephalopathies, such as lithium intoxication or as recently reported in acute Cefepime intoxication as well as with metabolic encephalopathies and rarely during AIDS. Periodic lateralized epileptiform discharges (PLEDs) have been classified according to their morphology and the associated condition (epileptic seizures, subtle status, "vascular" seizure). The principal etiology is cerebro-vascular disease and herpes encephalitis but it has also been reported in several other neurological diseases. PMID- 11915485 TI - Spike-wave discharge and the microstructure of sleep-wake continuum in idiopathic generalised epilepsy. AB - This review summarises all the evidences about the influence of different vigilance states on the occurrence of spike wave discharge (SWD) in idiopathic generalised epilepsy (IGE) patients. Numerous converging observations showed that full REM-sleep and alert wakefulness exert strong inhibition. A critical zone of vigilance which is a transitional state between waking and non-REM (NREM) sleep, and NREM sleep and REM sleep, has a promoting effect on the absence type spike wave discharge. Spike wave discharges are associated with phasic arousals without awakening and are attached to oscillation son the microstructural level of sleep, perpetuated by cyclic arousal events known as 'cyclic alternating pattern' (CAP), especially within the critical zone, but also along the whole sleep process. More specifically SWD seems to be attached to the 'A-phase' of CAP which is a reactive one and reflects synchronised NREM sleep EEG elements, like K-complexes, spindles and delta groups. The more slow wave elements are found in phase A--like in subtype A1--the more the coincidence with SWD occurs, and the more it is characterised by fast rhythms--as in subtype A2 and A3--the less the association with SWD could be observed. Since subtype A1 is associated with the first sleep cycle and with the descending branches of cycles, it is concluded that SWD appear in those dynamic moments of vigilance level oscillations which were characterised by strong sleep-like answers to arousal influences in high sleep pressure periods of sleep cyclicity. These data harmonize with another line of evidence suggesting that SWD represent the epileptic variant of the complex thalamocortical system function which is the substrate of NREM sleep EEG phenomena. In idiopathic generalised epilepsy there is a growing body of evidence that--as it was assumed by Gloor--spindles transform to SWD pattern. These data explain why those dynamic changes which evoke sleep responses are promoting for the occurrence of SWD. Adapting these data we offer a new interpretation to explain the strong activation effect of sleep deprivation in this kind of epilepsy. We assume that it is mainly due to the forced vigilance level oscillations, especially in morning, when elevated sleep pressure and circadian wake promoting forces, representing opposite tendencies, increase the amount of oscillations. PMID- 11915486 TI - Human alpha oscillations in wakefulness, drowsiness period, and REM sleep: different electroencephalographic phenomena within the alpha band. AB - Cortical oscillations in the range of alpha activity (8-13 Hz) are one of the fundamental electrophysiological phenomena of the human electroencephalogram (EEG). Evidence from quantitative EEG data has shown that their electrophysiological features, cortical generation mechanisms, and therefore, their functional correlates vary along the sleep-wake continuum. Specifically, spectral microstructure and EEG coherence levels between anterior and posterior cortical regions permit to differentiate among alpha activity spontaneously appearing in relaxed wakefulness with eyes closed, drowsiness period, and REM sleep, by reflecting distinct properties of neural networks involved in its cortical generation as well as a different interplay between cortical generators, respectively. Besides, the dissimilar spatiotemporal features of brain electrical microstates within the alpha range reveals a different geometry of active neural structures underlying each alpha variant or, simply, changes in the stability level of neural networks during each brain state. Studies reviewed in this paper support the hypothesis that two different alpha variants occur during human REM sleep: 'background responsive alpha activity', blocked over occipital regions when rapid eye movements are present, and 'REM-alpha bursts', non modulated by the alteration of tonic and phasic periods. Altogether, evidence suggests that electrophysiological features of human cortical oscillations in the alpha frequency range vary across different behavioural states, as well as within state, reflecting different cerebral phenomena with probably dissimilar functional meaning. PMID- 11915487 TI - Human brain potentials reveal similar processing of non-linguistic abstract structure and linguistic syntactic structure. AB - This research tested the hypothesis that shared or related neurophysiological processes are required for treating (a) non-linguistic abstract structure and (b) some aspects of linguistic syntactic structure. In language, one syntactic structure can be used to create an open class of sentences. We have previously proposed a relation between this generative aspect of syntactic structure and the abstract structure of non-linguistic sequences. For instance, the sequences 'ABCBAC' and 'DEFEDF' have different serial order or serial structure, but share the same abstract structure '123213'. Our recent studies of neuropsychology, simulation and ERPs argued that similar neurophysiological processes are involved in treating non-linguistic abstract structure and certain aspects of linguistic syntactic structure. The current research tests this hypothesis by examining the ERP profile evoked during the processing of non-linguistic sequences vs. sentences. Ten healthy subjects were trained to discriminate between syntactically correct and incorrect sequences and sentences presented visually on a video screen. During the subsequent ERP recording, subjects discriminated between correct and incorrect sequences and sentences presented visually on the screen. This discrimination task yielded, for anomalies in both the abstract and syntactic conditions, a late positivity around 550 ms with partially overlapping topography. These results support our hypothesis that shared or related neurophysiological processes are required for treating non-linguistic abstract structure and aspects of linguistic syntactic structure. However, they also suggest that the overlap between these two types of processing is not complete. PMID- 11915488 TI - Respective value of pudendal nerve terminal motor latency and anal sphincter electromyography in neurogenic fecal incontinence. AB - Fecal incontinence may be related to a neurogenic injury. Electrodiagnostic tests including pudendal nerve terminal motor latency (PNTML) and external anal sphincter electromyography (AEMG) have been proposed to reveal anal nerve damage. The aim of this study was to assess the respective value of PNTML and AEMG in the diagnosis of fecal incontinence. This study included 80 women (range 23-85 years) with fecal incontinence secondary to obstetrical and/or surgical trauma. They were evaluated by performing PNTML and AEMG. The electrophysiological results were compared and interpreted in the light of anorectal manometry (ARM) results. Electrodiagnostic test abnormalities were found in 64 of 80 patients (80%), including 28 patients with abnormal results for both tests and 36 patients with only one abnormal test. Overall, a neurogenic AEMG pattern was found in 64% of patients and a prolonged PNTML in 51%. No correlation was found between PNTML value and either AEMG grade or ARM parameters, while AEMG grade strongly correlated with squeeze pressure measured by ARM. This study showed that AEMG and PNTML did not give redundant information and allowed to explicit the mechanisms of neurogenic fecal incontinence. We found that AEMG was more sensitive and more closely related to the anal functional status (ARM parameters) than PNTML. These electrodiagnostic tests, particularly AEMG as performed in everyday practice, are useful in the assessment of neurogenic fecal incontinence. PMID- 11915489 TI - [Recent advances in childhood deafness]. PMID- 11915490 TI - [Study of psychological repercussions of 2 modes of treatment of adolescents with Crohn's disease]. AB - No study has yet compared the respective psychological impact of corticotherapy and enteral nutrition in the treatment of Crohn's disease, and especially, the psychological problems linked to the wearing of a nasogastric tube 24 hours a day. The goal of this study was to collect comparative information regarding the real-life experience and the feeling of these two treatments. PATIENTS AND METHODS: From September 1997 to February 1998 at the clinic of inflammatory bowel diseases of the hospital Necker-Enfants malades, 51 patients aged 12 to 18 (average 15) participated in this study. Thirty [15 on corticotherapy (CT); 15 on enteral nutrition (EN)] answered a questionnaire inspired by a similar Canadian questionnaire (Inflammatory Bowel Disease Questionnaire), and 21 passed Spielberger's anxiety tests, Beck's depression tests and a psychological interview. RESULTS: TREATMENT: According to the 30 questionnaires the appreciation of the therapeutic results was similar in the two treatments, the majority of patients respected their treatment (only one patient on EN ate secretly and two on CT stopped their corticotherapy). Of the 15 EN questionnaires: nine out of 15 patients responded well to the suspension of oral feeding, two were hungry, nine experienced cravings and ten avoided meals during their treatment. From a cosmetic point of view, six/14 (43%) found it difficult putting up with the nasogastric tube 24 hours a day, and eight/15 (53%) on CT found the facial swelling difficult to bear. According to the 21 psychological interviews, eight patients deemed EN efficient, while only four felt the same about CT. Of the 11 EN psychological interview, no adolescent patients were hungry, eight had cravings and nine avoided meals during their treatment, seven mentioned they felt different and seven described how EN had upset the family's routine. Nine (82%) talked about how difficult it was to put up with the nasogastric tube 24 hours a day from a cosmetic standpoint while eight/ten (80%) on CT found the facial swelling difficult to live with. Seven complained that they had been the victims of verbal abuse. Quality of life: According to the 30 questionnaires, eight/15 patients on EN missed an average of 15 days of school against five/15 patients on CT, ten patients judged that EN restricted their daily lives and nine mentioned the daily difficulties to wear a tube 24 hours a day. TESTS: Spielberger's test of anxiety revealed that on average, the 11 patients on EN who were interviewed suffered the first level of anxiety, while the ten patients on CT felt the second level. As for the Beck's depression test, the 11 patients on NE suffered the first level of depression on average, while those on CT were subject to the second level. CONCLUSION: This study demonstrated that EN was perceived as being more disruptive to patients daily lives than CT and defines the difficulties linked to the suspension of oral feeding and the wearing of a naso-gastric tube. The study also describes the difficulties involved in coping with the side effects of CT, one example being facial swelling which can be as unpleasant from a cosmetic point of view as wearing of a naso gastric tube. PMID- 11915491 TI - [Benefits of ipratropium bromide in the management of asthmatic crises in the emergency department]. AB - BACKGROUND: To determine if the addition of ipratropium bromide in the emergency department (ED) for the treatment of childhood asthma reduces rates of hospitalization and relapses for moderate and severe exacerbations. METHODS: Patients were given an oral corticosteroid treatment (2 mg/kg) and received every 20 minutes either three nebulizations with albuterol (0.15 mg/kg) and ipratropium bromide (250 micrograms) or six nebulizations with albuterol alone (control group). The primary end point was the need for hospitalization, additional nebulizations or a relapse during the following week. Secondary end point included the effect of age. RESULTS: One hundred and forty three children, two to 15 years old, were randomized to ipratropium or control groups and 121 were evaluated on day seven. As a whole, the control group was less often hospitalized or in relapse than those treated with three nebulizations of albuterol and ipratropium (17.5% vs 37.9%, p < 0.02). The ipratropium group reached the same result after three additional albuterol nebulizations. The benefit of anticholinergic therapy was observed for children less than six years of age who had a similar rate of success (73.5 vs 75.7%). CONCLUSION: The association of ipratropium bromide to the first three doses of the albuterol protocol for acute asthma did not act as well as six nebulizations of albuterol alone. The effect was age dependent and two to six years old children needed more attention. Nevertheless the hospitalization rate did not support the use of ipratropium compared with repeated albuterol nebulizations. PMID- 11915492 TI - [Hemophagocytic syndrome associated with neutropenia after chemotherapy]. AB - MATERIAL AND METHODS: This retrospective study reports 15 cases of hemophagocytic syndrome in children treated in our department during a eight-year period. RESULTS: Underlying diseases were acute lymphoblastic leukemia (n = 8) acute myeloblastic leukemia (n = 6) and Burkitt lymphoma (n = 1). Hemophagocytic syndrome was suspected after chemotherapy, in case of an unusual prolonged febrile neutropenia (n = 14) or isolated thrombocytopenia (n = 1). That fever was associated with cutaneous, pulmonary, hematologic, digestive and cardiac signs. Biological disorders included hypoprotidemia, hyponatremia, increased liver enzymes and fibrinopenia. Thrombocytopenia was observed in all patients and was associated with neutropenia for 14 of them. Diagnosis of hemophagocytic syndrome was always confirmed by bone marrow aspiration (infiltration with activated macrophages). Infection was documented in eight children. The treatment of hemophagocytic syndrome relied on steroids and resolution of symptoms occurred within three days of therapy. No recurrence of hemophagocytic syndrome was observed with a median follow up of two years and a half. CONCLUSION: Such complication should be suspected in cases of prolonged febrile neutropenia and/or thrombocytopenia, and confirmed by bone marrow aspiration. Indeed, steroid therapy is effective and chemotherapy can be then pursued. PMID- 11915493 TI - [Acquisition of secondary resistance after failure of a first treatment of Helicobacter pylori infection in children]. AB - AIMS: To assess the frequency of acquisition of secondary Helicobacter pylori resistant-strains after a first course of antimicrobial treatment. PATIENTS AND METHODS: A retrospective study was performed during the 1994-2000 period, in 15 girls and eight boys, mean age 10.9 +/- 4.8 years (1.4-17 years), with Helicobacter pylori gastritis (culture and antimicrobial susceptibility) presenting a failure of first course treatment, with during one week a proton pump inhibitor and amoxicillin together with either clarithromycin (n = 14) or metronidazole (n = 9). Two endoscopies were performed, the first at the time of diagnosis and the second after the failure of bacterial eradication demonstrated by a positive 13C urea breath test six weeks after the end of treatment. Antimicrobial susceptibility of all Helicobacter pylori strains was tested after each endoscopy and before starting a second course of the treatment. RESULTS: Comparison of antimicrobial susceptibility before and after the first course of treatment showed that Helicobacter pylori strains were all sensitive to amoxicillin, clarithromycin-resistant in eight children (34.7%) before treatment vs 12 (52.1%) after treatment, p = 0.42, ns, metronidazole-resistant in 13 (56.5%) vs 12 (52.1%), p = 0.80, ns, and both clarithromycin and metronidazole resistant in four (17.3%) vs seven (30.4%), p = 0.63, ns. Among the 14 children treated by a triple therapy including clarithromycin, three (21.4%) developed a secondary resistance to clarithromycin and in one metronidazole resistance was no more detected. Among the nine children treated with a triple therapy including metronidazole, none developed a secondary resistance to metronidazole and one developed a secondary resistance to clarithromycin. CONCLUSION: This study shows the absence of amoxicillin-resistant strains, a high initial clarithromycin resistant strains level (primary resistance), increasing after a first course of treatment, and for metronidazole a high initial level of resistance not influenced by treatment. Secondary clarithromycin-resistance of Helicobacter pylori strains following the first course of treatment could account for failure of bacterial eradication and suggests the importance of antimicrobial susceptibility. PMID- 11915495 TI - [Primary Gougerot-Sjogren syndrome in a 13-year-old girl]. AB - Sjogren's syndrome is uncommon in children, and occurs most often in association with autoimmune diseases (secondary Sjogren's syndrome). We describe the clinical and biological features of a 13-year-old girl with primary Sjogren's syndrome, revealed by recurrent parotitis. CASE REPORT: This adolescent girl was referred for investigation of multiple episodes of bilateral parotid swelling since age nine, without systemic symptoms. Examination was unremarkable except for enlarged and painless parotid glands. Laboratory investigations, measurement of saliva production, parotid sialography, labial salivary gland biopsy, revealed Sjogren's syndrome without associated disease. Hydroxychloroquine was prescribed with clinical improvement. CONCLUSION: Recurrent parotitis in children is an uncommon condition. The onset of parotid swelling at five years or over deserves screening for dysimmune disorders, sarcoidosis or Sjogren's syndrome. Diagnosis of Sjogren's syndrome is based on laboratory evidence of autoimmune disorders and minor salivary gland biopsy. PMID- 11915494 TI - [Acute abdominal pain in children at the Pediatric Hospital in Bangui (Central African Republic). Epidemiological, clinical, paraclinical, therapeutic and evolutive aspects]. AB - Acute abdominal pains in children in general, and in subsaharian Africa in particular, are among the most frequent causes of consultations. MATERIAL AND METHODS: The authors achieved a two-year retrospective study on acute abdominal pains in children, with the objectives of determining the frequency of acute abdominal pains, precising the epidemiological, clinical, therapeutic and evolutive aspects and different aspects of the treatment. The study concerned 312 cases. RESULTS: A predominance of the ages three to ten years was noted (67% of the cases). Most of the patients were related to low income parents. Half patients were submitted to antiparasitical treatment before arriving at the hospital and they were treated mainly by antihelminthiasis. The diagnosis were appendicitis (32.4%), typhoid perforations (9.9%), digestive forms of acute malaria (5.8%), strangulated herniae (4.8%), acute gastroenteritis (4.8%), acute pneumoniae (3.8%), urinary tract infections (3.5%), amibian liver abscess (1.9%), viral hepatitis (1.6%), Schonlein Henoch purpura (1.3%), occlusive syndrome (1.3%) and other medical causes (2.2%). In 22.8% of the cases, no cause was found. CONCLUSION: The ignorance of the seriousness signals, late recourses to hospitals structures and the limited financial means explain the high rate of mortality (8%), mainly due to appendicitis and typhoid perforations. PMID- 11915496 TI - [A case of atrial flutter after umbilical venous catheterization]. AB - Isolated atrial flutter is an extremely rare form of supraventricular tachycardia in the neonatal period. It may be initiated by central venous catheterization. CASE REPORT: A male infant was born at 35 weeks by cesarean section for placenta praevia. He was eutrophic. Apgar score was 10 at 1 and 5 minutes. He secondary developed a respiratory distress syndrome. He was then ventilated by nasal CPAP. Immediately after an umbilical venous catheterization, a tachycardia appeared without preexistent cardiac dysfunction. An intravenous dose of adenosine (Striadyne) showed a characteristic sawtooth pattern of P waves on inferior leads. The cardiac-US examination was normal. This atrial flutter was converted to normal sinus rhythm by transoesophageal pacing, without adjunction of antiarrhythmic drugs. The newborn was weaned from mechanical ventilation 48 hours later and discharged from hospital at seven days post natal age. His development and clinical examination were normal two months later. CONCLUSION: The isolated atrial flutter is rare in the neonate. It may be triggered by a venous catheterization. Transoesophageal atrial pacing is safe and effective for conversion. PMID- 11915497 TI - [Acute intestinal invagination revealing celiac disease in a 9-month-old infant]. AB - In most cases bowel intussusception is idiopathic in children. Indirect causes, such as celiac disease, are rare. CASE REPORT: A nine-month-old girl was admitted for a bowel intussusception diagnosed by ultrasonography, yet not confirmed by barium enema. The girl underwent a thorough check up, due to a loss of weight and denutrition symptoms which evidenced immunoglobulin A anti-gliadin, immunoglobulin G anti-gliadin, immunoglobulin A anti-endomysium and antireticulin positive antibodies. Celiac disease was confirmed by bowel biopsy, which revealed a subtotal villous atrophy. Evolution was favourable under a gluten free diet. COMMENTS: The description of a bowel intussusception associated with celiac disease is common. This observation appears to be the earliest case ever described: the patient was only nine months old. The initial procedure is routine: diagnosis by ultrasonography, therapeutic enema and surgery only in case of failure of medical treatment. Intussusception associated with celiac disease often presents in an atypical way: elementary forms, spontaneously resolvent and recidivious. The knowledge of this clinical set of symptoms leads to an early diagnosis of celiac disease. The gluten free diet prevents the recurrence of intussusception. PMID- 11915498 TI - [Staphylococcal toxic syndrome, atypical presentation of Kawasaki syndrome or staphylococcal skin syndrome?]. AB - CASE REPORT: A three-year-old girl was admitted for persistent fever, erythermatous rash with subsequent desquamation, stomatitis, cheleitis and cervical lymphadenopathy following development of a buttock abscess secondary to an insect bite. A TSS-positive Staphylococcus aureus strain was isolated from the abscess. COMMENTS: Both clinical and bacteriological features led to discuss a "toxic shock syndrome without shock", an atypical form of Kawasaki syndrome without thrombocytosis and coronary arteritis or a staphylococcal skin syndrome. An early treatment with antibiotics could have limited the toxin production explaining both symptomatology and favourable course of the disease. PMID- 11915499 TI - [Bronchopulmonary dysplasia and corticosteroid therapy]. AB - Inflammation plays a major role in the pathogenesis of bronchopulmonary dysplasia of preterm infants. Having a strong anti-inflammatory effect, corticosteroids have been proposed in the management of this disease. Antenatal steroids protect the newborn against its development. Postnatal systemic administration of steroids reduces the duration of mechanical ventilation and oxygen therapy, but is associated with short term and long term adverse effects. Early administration of dexamethasone (before 7 days of life) reduces the incidence and the severity of chronic lung disease at 28 days of life and 36 weeks of post-conceptional age. Inhaled steroids are associated with less adverse effects than dexamethasone administration, but they are also less effective. PMID- 11915500 TI - [Radiological case of the month]. PMID- 11915501 TI - [What's new in fetal medicine?]. AB - One of the major progress in fetal medicine in recent years is the increased sensitivity of sonographic screening for foetal malformations, due to technical improvement but also to a better training of professionals. Screening for chromosomal abnormalities is no longer based on maternal age alone. Second trimester maternal serum screening (MSS) is increasingly used: thus in 1997, 376,798 MSS tests were performed in France, yielding to the prenatal diagnosis of 391 cases of Down's syndrome. First trimester sonographic nuchal translucency measurement (NTM) is an effective screening method when performed under stringent conditions. Quality control however, is more difficult to implement on a large scale for NTM than for MSS. Performing screening tests sequentially carries a danger of generating an unnecessarily high number of amniocentesis, which may be obviated by a rational calculation of an individual's risk to carry an aneuploid baby. First trimester MSS is expected to become standard practice in the next years, probably in combination with NTM. Cytogenetics underwent substantial innovations recently, due to the ever-increasing use of molecular cytogenetics. FISH techniques allow: 1) precise analysis of unexpected structural chromosomal abnormalities diagnosed by routine amniocentesis, 2) rapid screening of the most common aneuploidies by amniocentesis when a fetal structural anomaly is detected by 3rd trimester ultrasound, 3) diagnosis of micro-deletions suspected by fetal ultrasound or post-mortem. Prenatal diagnosis by maternal blood sampling and fetal cells or DNA analysis is now part of routine clinical practice in selected cases, such as fetal sexing in families affected by an X linked disease. Thus one can select those pregnancies eligible to invasive prenatal diagnosis. Pre implantation diagnosis, which has not been legal in France until 1999 is now increasingly used as an alternative to first trimester diagnosis. As for fetal therapy, a major recent breakthrough is the prenatal management of twin to twin transfusion syndrome by either amnioreduction or laser coagulation of inter-twin vascular shunts. In addition, new pathophysiologic concepts involving the renin angiotestin system could lead to further therapeutic innovations. A European randomised trial is now being completed to establish the respective indications of drainage and Laser. All this underscores that fetal medicine is no longer solely a succession of dramatic technical breakthroughs, but is entered an era of large-scale diffusion that requires evidence based evaluation. PMID- 11915502 TI - [Adolescents and contraception. What should the pediatrician know? (Part 2)]. AB - Many adolescents are sexually active and reproductive health is an important aspect of adolescent medicine. However, pediatricians are often uncomfortable with the issues of sexuality and contraception, for which they have not been particularly trained. The general purpose of this article is to increase pediatricians' sense of competence with adolescents, particularly when having to deal with or counsel on such a sensitive issue as contraception. This second of two parts addresses confidentiality and prescription of oral contraceptives to minors, how to ask questions about sexual and reproductive life, best initial choice and eventual subsequent switch of contraceptive pill, acne and oral contraception, problems with mental handicap, and finally, what about boys. Pediatricians interested in the care of adolescents are strongly invited to keep developing their expertise in the field of sexuality, contraception and reproductive health. PMID- 11915503 TI - [Bidirectional cell traffic between mother and fetus: new insights in pediatric and adult disorders]. AB - There is a heavy traffic of cells and DNA through the placenta during pregnancy. The rate of fetal cells in the maternal blood is correlated with abnormalities, such as aneuploidy and pre-eclampsia. Studying and quantifying these cells could improve antenatal diagnosis techniques, especially for Down syndrome. Maternal fetal microchimerism is frequently observed in several auto-immune diseases in adulthood, such as systemic scleroderma. Studies suggest a rather allo-immune pathophysiology, involving maternal-fetal HLA compatibility. Microchimerism is also found in auto-immune diseases in children. Thus, the cells traffic offers new insights for antenatal diagnosis techniques and pathophysiology of auto immune diseases. PMID- 11915505 TI - [Kawasaki disease and dengue: a fortuitous association?]. PMID- 11915504 TI - [Congenital toxoplasmosis: prevention in the pregnant woman and management of the neonate]. AB - The management of a pregnant women or a child infected by Toxoplasma gondii rests on the screening of pregnant women at risk of infection. Treatment is prescribed if an infection occurs. A prenatal diagnosis (detection of T. gondii in amniotic fluid) may be performed, a positive result leading to a reinforcement of the treatment. After birth, the follow-up of the child is needed in order to prevent and detect the sequellae, mainly ocular. For all these steps, the biological methods used to diagnose T. gondii infection are of paramount importance. PMID- 11915506 TI - [Endoscopically discovered pancreatic heterotopia: intervention or not?]. PMID- 11915507 TI - [Prevention of community-acquired respiratory syncytial virus infections in premature infants: cost-benefit evaluation in the Finistere Department]. PMID- 11915508 TI - [Legg-Perthes-Calve disease and disorders of hemostasis]. PMID- 11915509 TI - [Neonatal detection of deafness by molecular genetics in maternity hospitals: report of a 1st year concerning families at risk]. PMID- 11915510 TI - Drug use benchmarks show surprising geographic variation. PMID- 11915511 TI - Study these variables to understand reinsurance needs. PMID- 11915512 TI - Simple formula shows impact of changes in RVU conversion factor. PMID- 11915513 TI - Poor alignment, unreliable data frustrate PHO in low managed care market. PMID- 11915514 TI - Know your costs? Check these benchmarks for comparison. PMID- 11915515 TI - [Functional alterations of astroglia on brain pathologies and their intracellular mechanisms]. AB - A phenotypic alteration of astroglia, "astroglial activation", is a common phenomenon observed on brain pathologies. The hypertrophy/hyperplasia of activated astroglia causes a glial scar, which prevents synaptic re-generation. In contrast, many neurotrophic substances are produced by the activated astroglia. Thus, the functional alteration of astroglia is important in tissue repair processes of the damaged CNS. Endothelins (ETs) are involved in the pathophysiological responses of the CNS. We found that injection of ETs into rat brain induced activated astroglia. A selective ETB-receptor antagonist attenuated the induction of activated astroglia. In cultured astroglia, ETs reproduce the functional alterations characterizing activated astroglia; i.e., increases in proliferation, morphological changes and stimulation of several gene transcriptions. ETs re-organized astroglial cytoskeletal actin through a small GTP-binding protein, rho, which may underlie the astroglial hypertrophy. Analysis of gene expression showed that transcriptions of neurotrophic factors (GDNF and BDNF) were stimulated by ETs. ETs stimulated astroglial proliferation by both adhesion-dependent and -independent mechanisms, where FAK and ERK plays key roles, respectively. These findings suggest important roles of ETs in the regulation of astroglial functions. PMID- 11915517 TI - [Functional analysis of calcineurin-mediated signalling pathway using fission yeast as a model system]. AB - Calcineurin (CN), a highly conserved Ca2+/calmodulin-regulated phosphatase, is a critical component of many calcium-regulated processes in mammalian cells, including T cell activation, cardiac hypertrophy, learning and memory. CN is specifically inhibited by the immunosuppressant drugs cyclosporin A and tacrolimus (FK506), and these drugs have served as valuable reagents in identifying the role of CN in a wide variety of cell types. CN may have additional functions in other cell types, and the loss of these functions may contribute to the side effects of these drugs, which include nephrotoxicity and neurotoxicity. A better understanding of the biological roles of CN in different cell types may promote the development of improved strategies for immunosuppression. We have been studying the CN signal transduction pathway in fission yeast because this system is amenable to genetics and has many advantages in terms of relevance to higher systems. Fission yeast has a single gene encoding the catalytic subunit of CN, ppb1+, that is essential for cytokinesis. We have shown that in fission yeast CN plays an essential role in maintaining chloride ion homeostasis and acts antagonistically with the Pmk1 MAP kinase pathway. We also carried out an isolation and a screening for several FK506-sensitive mutants in order to identify genes that share an essential function for viability with CN. Possible roles of these gene products in cellular functions in relation to calcineurin are discussed. PMID- 11915516 TI - [Molecular identification of LOX-1 and analysis of its pathophysiological role]. AB - Recent progress in the study of atherosclerosis revealed that the proatherogenic property of LDL is attributable to oxidized LDL. Macrophages recruited to vascular wall phagocytose oxidized LDL and transformed into foam cells, which is a hallmark of atheroma. Endothelial cells also binds oxidized LDL and changes its phenotype to the status of "endothelial dysfunction." We successfully cloned the endothelial receptor for oxidized LDL, designated LOX-1. LOX-1-mediated action of oxidized LDL induces the decrease in NO release and the increased expression of adhesion molecules, which are typical changes in endothelial dysfunction. The expression of LOX-1 is quite inducible. Proinflammatory cytokines, etc. induce the expression of LOX-1 in vitro; and proatherogenic conditions, e.g., hypertension, hyperlipidemia, and diabetes, induce the expression of LOX-1 in vivo. This manner of expression suggests the importance of LOX-1 in pathological settings. LOX-1 binds not only oxidized LDL, but also binds apoptotic cells and activated platelets through the interaction with anionic phospholipids. This property might bridge atherosclerosis and thrombosis. A novel system to detect LOX-1 ligand in plasma detected the increased level of LOX-1 ligand in hypercholesterolemic rabbits compared with normal ones. This system might be useful to predict the status of endothelial function and the risk of ischemic heart disease. PMID- 11915518 TI - [Culturing hippocampal neurons]. AB - The procedure for making a low density culture of hippocampal neurons has been elaborated by Goslin and Banker. The viability of hippocampal neurons, which are sparsely disseminated on the glass surface, is maintained by a separately cultured glial monolayer; the glial feeder layer is grown on the bottom surface of the dish, while those neurons, placed face down, are attached on the coverslips. This method is originaLly designed for the observation of the maturation, polarity and axogenesis of a single neuron. In addition, this method can be applied for a variety of other purposes: (1) to observe synaptogenesis, (2) to analyze synaptic function electrophysiologically, (3) to analyze receptor functions and signaling cascades pharmacologically, (4) to visualize a molecular dynamics by time-lapse analyses of GFP-tagged molecules, and (5) to observe ultrastructure by an electron microscope. Furthermore, these neurons are useful even in biochemical experiments because they are relatively uniform without glial contamination and highly enriched in synaptic components. PMID- 11915519 TI - [Hair growth effect of minoxidil]. AB - The length and size of hair are depend on the anagen term in its hair cycle. It has been reported that the some cell growth factors, such as VEGF, FGF-5S, IGF-1 and KGF, induce the proliferation of cells in the matrix, dermal papilla and dermal papillary vascular system and increase the amount of extra cellular matrix in dermal papilla and then maintain follicles in the anagen phase. On the other hand, negative factors, like FGF-5, thrombospondin, or still unknown ones, terminate the anagen phase. If the negative factors become dominant against cell proliferation factors according to fulfilling some time set by the biological clock for hair follicles, TGF beta induced in the matrix tissues evokes apoptosis of matrix cells and shifts the follicles from anagen to catagen. Androgenetic alopecia is caused by miniaturizing of hair follicles located in the frontal or crown part of scalp and are hereditarily more sensitive to androgen. In their hair cycles, the androgen shortens the anagen phase of follicles and shifts them to the catagen phase earlier than usual. The mode of action of hair growth effect of minoxidil is not completely elucidated, but the most plausible explanation proposed here is that minoxidil works as a sulfonylurea receptor (SUR) activator and prolongs the anagen phase of hair follicles in the following manner: minoxidil (1) induces cell growth factors such as VEGF, HGF, IGF-1 and potentiates HGF and IGF-1 actions by the activation of uncoupled SUR on the plasma membrane of dermal papilla cells, (2) inhibits of TGF beta induced apoptosis of hair matrix cells by opening the Kir 6.0 channel pore coupled with SUR on the mitochondrial inner membrane, and (3) dilates hair follicle arteries and increases blood flow in dermal papilla by opening the Kir 6.0 channel pore coupled with SUR on the plasma membrane of vascular smooth muscle cells. PMID- 11915520 TI - [Pharmacological and clinical properties of levocabastine hydrochloride (eye drop and nasal spray), a selective H1 antagonist]. AB - Levocabastine is a selective histamine H1-receptor antagonist exerting inhibitory effects on the release of chemical mediators from mast cells and on the chemotaxis of polymorphonuclear leukocytes and eosinophils. Both histamine and antigens induced conjunctivitis was inhibited by levocabastine in several allergy models. Levocabastine moderately inhibited histamine-release from guinea pig conjunctive induced by antigen-antibody reactions and prevented an increase in the vascular permeability of the conjunctive elicited by both histamine and antigen instillation. Symptoms of allergic rhinitis, which were induced by histamine, substance P and antigen, were also reduced by levocabastine. Levocabastine prevented an increase in the vascular permeability of nasal mucosa elicited by instillation of these three inducers. Furthermore, levocabastine has shown a large difference between the antiallergic dose and other non-specific pharmacological effective dose than that with other antiallergic drugs. The non specific pharmacological effect of levocabastine reveals only blepharoptosis. With these pharmacological effects and topical usage, levocabastine was shown to be useful for allergic conjunctive and rhinitis in both seasonal and perennial clinical use. PMID- 11915521 TI - [Physicochemical and pharmacological characteristic and clinical efficacy of an anti-irritable bowel syndrome agent, polycarbophil calcium (Polyful)]. AB - Irritable bowel syndrome (IBS) is a functional bowel disorder characterized by abdominal pain or discomfort and abnormal defecation. Polycarbophil calcium, a water-absorbing polymer, is expected to improve stool consistency. Polycarbophil calcium decalcified under the acidic condition and then absorbed 70 times its weight of water under the neutral condition. In in situ experiments using rat jejunum and colon, polycarbophil decreased water absorption by the intestine without affecting water secretion. Polycarbophil inhibited prostaglandin E2-, 5 hydroxy-L-tryptophan- and castor oil-induced diarrhea in mice or rats. Polycarbophil calcium also inhibited sennoside-induced diarrhea in dogs. Polycarbophil increased the weight of feces in naive or low-fiber diet feeding rats. In naive dogs, polycarbophil calcium increased stool frequency, stool weight and moisture. Polycarbophil was not absorbed from the gastrointestine, not metabolized and eliminated into feces in rats and dogs. Polycarbophil calcium did not affect the absorption of coadministered drugs in dogs. In the dose-finding clinical study for IBS, polycarbophil calcium was effective both in diarrhea and constipation. In the Phase III study, polycarbophil calcium was superior to trimebutine maleate in efficacy and equal in safety. Emesis/vomiting and thirst were observed, but episodes of diarrhea or constipation by excessive action were few. Polycarbophil calcium seems promising as an anti-IBS agent. PMID- 11915522 TI - Was it just Shelly? PMID- 11915523 TI - When few were women. PMID- 11915524 TI - The battle for balance. PMID- 11915525 TI - Health care for him. PMID- 11915526 TI - Barriers to disclosure of abuse among rural women. AB - The purposes of this study were to examine the prevalence of abuse (physical, emotional, and sexual) in women seeking care in rural medical clinics and WIC voucher pick-up sites, and to discover ways of improving the response of health care providers to violence. Data were collected in 8 medical clinics and 17 WIC supplemental food program sites in 9 counties of west central Minnesota during January and February 1997. Fifteen percent of respondents reported having had a discussion about abuse with a health care provider. Six of the 8 symptoms and injuries most associated with abuse indicate diminished emotional health. A series of barriers are identified as substantial obstacles to obtaining help and revealing abuse to health care providers. The most frequently reported barriers were self-reliance, reliance on God, and reliance on friends and family. These findings show that a large percentage of rural women experience abuse and that their health is adversely affected. The barriers to disclosure of abuse reported in this study illustrate the complexity of disclosing abusive relationships in rural and other settings. Low screening levels suggest that rural health care providers can develop additional opportunities to discuss abuse with their patients. PMID- 11915527 TI - Exploring the dual-physician marriage. AB - A substantial number of female physicians marry fellow doctors, yet little is known about these dual-physician couples. In an effort to identify these couples, surveys were sent to 1,695 female physician members of the Minnesota Medical Association. Women who were or had been married to a physician were asked to complete the survey. The majority of women responding (n = 203) were between the ages of 36 and 45 and had married during medical school. Only a small percentage (11.3%) were divorced, and medicine was reported to play a role in 69.6% of those separations. Questions were asked regarding work and family life, and job satisfaction levels. Despite many positive responses to the questionnaire, some problems existed in these marriages. Overall, however, responses indicated that the advantages of being married to another physician for outweigh the disadvantages. PMID- 11915528 TI - Gender-based harassment. PMID- 11915529 TI - The electronic medical record. Good-bye paper charts, hello better patient care. PMID- 11915530 TI - Surgery for gastroesophageal reflux disease. PMID- 11915531 TI - Toward a better mousetrap. PMID- 11915532 TI - First and foremost a surgeon. PMID- 11915533 TI - Improve, don't reject AMA. PMID- 11915534 TI - Trust the doctor's judgment. PMID- 11915535 TI - Stalking the no. 1 killer. Texas physicians attack cardiovascular disease on multiple fronts. PMID- 11915536 TI - Sick doctors. PMID- 11915537 TI - Point of no return. Resitution disappoints doctors, prompts aggressive insurers' reaction. PMID- 11915538 TI - Catch 22 renumbered. Texas physicians frustrated by clean claim requirements. PMID- 11915539 TI - Define 'enough'. TMA questions the need for new medical schools. PMID- 11915540 TI - A citywide evaluation of identification of risk factors for cardiovascular disease in emergency department patients complaining of chest pain. AB - The local division of the American Heart Association in Lubbock, Tex, conducted a cooperative study involving all three local hospitals to assess how effectively risk factors for cardiovascular disease are identified in patients presenting in the emergency departments with a complaint of chest pain. The charts of 250 consecutive patients with chest pain were reviewed for risk factors identified by the triage nurse and by the emergency department physician or the attending physician or both. Although the rate at which risk factors were identified was good, identification was neither complete nor comprehensive. Comparison with statistics for the general population showed that some risk factors were over reported, while others were markedly underreported. Each of the three specialties of health care professionals stressed different risk factors, and having all three involved markedly increased the level of identification. The data provide guidelines for improving risk factor identification, and the study led to the formation of strategic alliances among the different hospitals and health care professionals that should help improve secondary prevention of cardiovascular disease within the community. PMID- 11915541 TI - Recognizing and managing parafunction in the reconstruction and maintenance of the oral implant patient. AB - A primary goal in the reconstruction of the dental implant patient is to achieve the lowest stress possible on the abutments, both artificial and natural. Bidez and Misch use the formula [figure: see text] [formula: see text] to explain that the stress can be easily lowered by increasing "A," by using larger implants, or by using additional implants. Another equally important method of lowering the stress is to reduce and manage the force application (F). The majority of completed implant cases can well tolerate applied forces from mastication and swallowing, both of which are close to 50 pounds per square inch, but it is the destructive power from DCS that can easily range from 500 to 1,000 pounds per square inch with which this article is concerned. PMID- 11915542 TI - Fracture and fatigue behavior of shot-blasted titanium dental implants. AB - This investigation studies the effect of the shot-blasting treatment on the cyclic deformation behavior of a commercially pure titanium, with two microstructures: equiaxed and acicular. The fatigue tests were carried out in artificial saliva medium at 37 degrees C. Cyclic deformation tests have been carried out up to fracture, and the fatigue crack nucleation and propagation have been analyzed. The results show that the shot-blasting treatment improves the fatigue life in the microstructures studied, and that the equiaxed was better in mechanical properties than the acicular. The cause of this improvement in the mechanical properties is due to the compressive stress on the material surface for the shot-blasted specimens. Hardness tests were carried out to determine the value of these internal stresses. PMID- 11915543 TI - Vertical dimension of occlusion in implant dentistry: significance and approach. AB - Understanding the principles of occlusion as they relate to managing bite forces on implants is extremely important to maintain the longevity of dental implant prostheses. Specifically, altering vertical dimension of occlusion (VDO) scientifically and predictably is critical to the creation of proper tooth form and guidance when fabricating a full-arch implant-supported prosthesis. PMID- 11915544 TI - Evolution of the concept of angulated abutments in implant dentistry: 14-year clinical data. AB - This paper describes the evolution of the concept of selecting the abutment at first-stage surgery and presents clinical data accumulated over 14 years of the use of this concept with angulated abutments. The concept was developed for implants using internal and external hexed connections and has evolved in its use for implants using a Morse taper connection as a result of the considerable clinical advantages that this type of connection offers. A total of 3101 implants were restored using angled abutments ranging from 0 to 45 degrees and were observed over a period of up to 151 months. After an observation time of 120 months (10 years) after placement, the calculated 95% confidence interval of the mean survival estimation, according to Aalen et al, was 98.2% (+/- 0.7%). The magnitude of the angles did not influence the survival rate. The need to refine the implant position in terms of the depth to which it was placed and the angulation and rotational orientation, as well as the size and pivot point of the final abutment, were recognized as contributing to the harmonious emergence profile of the restoration. Good esthetic and functional outcomes were achieved by the use of conventional cement-retained restorations made possible by parallel and aligned abutments. PMID- 11915545 TI - Enhancement of primary stability of dental implants using cortical satellite implants. AB - PURPOSE: This study aims to assess the effect of satellite implants on the primary stability of dental implants placed in fresh extraction sites in vitro. METHOD: 34 titanium screw implants (3.75 mm x 10 mm; Bego, Bremen, Germany) were inserted in premolar- and molar-fresh extraction sites in domestic pig mandibles. Periotest (PT) values were assessed before and after insertion of one vestibular and one lingual 1.7-mm bone screw (Mondeal, Tuttlingen, Germany) as a satellite implant was connected to the implants with a 0.6-mm microplate welded to the implant abutment. RESULTS: The average PT values were 2.9 without satellite implants, -1.0 with one satellite implant, and -2.5 with two satellite implants during horizontal testing, and 3.0, 1.4, and 0.4, respectively, for vertical testing. CONCLUSION: Satellite implants increase the horizontal stability of implants in fresh extraction sites. Differences for horizontal PT assessment were significant on a 0.01 level of confidence. Implants in extraction sites may be loaded immediately, if vertical stabilization is provided by cortical bone and if horizontal PT values show sufficient stability after satellite implant insertion. PMID- 11915546 TI - Analysis of 435 screw-vent dental implants placed in 161 patients: software enhancement of clinical evaluation. AB - This paper demonstrates how a computer software program was utilized in a private practice to supplement the clinical evaluation of one implant system. Clinical data were entered into a computer database at the time of implant placement and up to 13 years for follow-up appointments. Data were divided into two groups and subjected to lifetable analyses. The focus group consisted of a machined titanium, screw-type implant with an internal abutment connection from one manufacturer. The residual database consisted of mixed implant designs with a variety of abutment connections and surfaces from several other manufacturers. Lifetable survival data between the two groups were generated. Cumulative survival rates from 0 to 13 years were 94.2% (n = 435) for the focus group and 90.1% (n = 2339) for the reference group. There were 25 implants lost in the focus group and 11 other implants were deemed "at risk." Survival results from other lifetable analyses are also presented for the two groups. Documentation of empirical clinical data in a computer software database over a period of time can help private practice clinicians better evaluate the dental implant systems used in their practices. PMID- 11915548 TI - European markets for dental implants 2001: executive summary. PMID- 11915547 TI - Risk management when operating in the posterior mandible. AB - A risk management protocol for the placement of implants in the posterior mandible is a sensible precaution for reducing the risk of nerve damage and the risk of possible litigation. This paper identifies the need for careful treatment planning and the use of relevant tests (e.g., the CT scan). A protocol is presented that advised the use of certain procedures to reduce the risk of nerve damage. These procedures consider the incision, the type of anesthetic, x-rays as you proceed, the type of implant, and use of anti-inflammatories. PMID- 11915550 TI - Simplified technique for immediate implant insertion into extraction sockets: report of technique and preliminary results. AB - A technique is presented for placement of implants at the time of tooth extraction in the precisely desired positions, regardless of extraction socket morphology. To date, 162 implants have been placed utilizing this technique. One was mobile at uncovery and removed. The other implants have been successfully in function, as defined by the Albrektsson Criteria, for up to 7 years, yielding a cumulative success rate of 99.4%. PMID- 11915549 TI - A concept for the treatment of various dental bone defects. AB - Untreated dental bone defects usually lead to resorption of alveolar bone. Filling these defects with bone substitute material prevents resorption of bone, preserves the alveolar ridge, and provides sufficient bone for immediate or subsequent implant placement. A variety of bone substitutes is available. They differ in origin, consistency, particle size, porosity, and resorption characteristics. We have treated almost 1000 bony defect sites in 267 patients with the bone regeneration material Cerasorb. Being resorbed simultaneously with the formation of new bone, it is completely replaced by the patient's own vital bone within 6 to 12 months. The representative cases described in this paper demonstrate the successful use of the pure-phase beta-tricalcium phosphate ceramic in the treatment of all dental bone defects. PMID- 11915551 TI - Diabetes mellitus and dental implants: a clinical study. AB - Diabetes mellitus is no longer considered to be a contraindication for implant supported prostheses, provided that the patient's blood sugar is under control, and that there is motivation for oral hygiene procedures. This paper presents the experiences of treating diabetic patients using implants with good success rates. PMID- 11915552 TI - Immediate placement in extraction sites followed by immediate loading: a pilot study and case presentation. AB - The purpose of the study was to compare the results of implants immediately loaded in edentulous sites with implants loaded immediately in extraction sites. Since December 1998, we selected a small group of patients for immediate or early loading. Seventy-five implants were placed in nine jaws of seven patients. Two of the patients received implants in both the maxilla and mandible. Of the 75 implants placed, 29 were placed in immediate extraction sites. Twenty-six of the 29 that were placed in immediate extraction sites were loaded in less than 3 weeks. Of the 75 implants placed, 62 were loaded early (less than 3 weeks). Two implants have been lost. The remaining 13 implants were buried and allowed to heal in the customary manner. None of the buried implants failed. One of the implants lost was in an extraction site and one was in a nonextraction site. Of the 33 implants that were placed in edentulous areas and immediately loaded, one was lost. This is compared with the 29 implants placed in extraction sites that were immediately loaded. We conclude that the success rates for implants immediately loaded in extraction sites and edentulous sites are comparable. PMID- 11915553 TI - [Molecular cancer therapy using tyrosine kinase inhibitors]. PMID- 11915554 TI - [Thermometry]. PMID- 11915555 TI - [Mechanisms of fever]. AB - Fever represents a normal and adaptative response developed in case of host aggression, notably by infectious agents, and is part of a large defense system, the acute phase response. Fever mediators are mainly derived from the host cells and are pyrogenous cytokines such as interleukin-1, tumour necrosis factor alpha, interleukin-6 or interferons which act at the hypothalamus level via prostaglandin E2. Although fever has been conserved through evolution, its beneficial effects in humans are not well-established. PMID- 11915556 TI - [Intermittent fever of infectious origin]. AB - Intermittent fever is rare during the course of infectious diseases but it represents a diagnostic and therapeutic challenge. The most frequent infectious causes of intermittent fever are focal bacterial infections, mainly infections localised to canals like urinary or biliary ducts or the colon and also infections of a foreign material. Other causes are less frequent, like infective endocarditis, tuberculosis, infections due to Yersinia enterocolitica or malaria, or exceptional like borreliosis, ratbite fever, chronic meningococcemia or chronic Epstein-Barr Virus infection. Careful anamnesis and clinical examination as well as a few simple complementary investigations, preferably performed during a febrile episode, are often sufficient to set the limits of possible further more complex investigations. PMID- 11915557 TI - [Intermittent fever of neoplastic origin]. AB - Cancer and hemopathy account from 4.5% to 14% of causes of intermittent fever (IF). The tumoral etiology is loss frequent in IF than continues fever. The man physiopathologic mechanism is represented by secretion of cytokines from tumour cells. IF may be the presenting symptom of cancer. This case diagnosis is usually made by cautions clinical evaluation and imaging. When cancer is known, IF can be related to infectious diseases or neoplastic evolution. PMID- 11915558 TI - [Familial Mediterranean fever]. AB - Familial mediterranean fever is a hereditary inflammatory disease, with autosomal recessive transmission, due to mutations in the MEFV gene. The MEFV gene, located on the short arm of chromosome 16, codes an anti-inflammatory protein, marenostrine or pyrin. The disease is characterised by paroxysmal bouts of fever with acute and painful serositis. Appearance of renal amyloidosis indicates severe prognosis. The disease appeared several thousands of years ago in an ancestor common to Sephardic Jews, Turks, Armenians and Arabs. The full clinical description, including renal complications and familial forms, was made by two French investigators and dates from the 1950s. That this description is relatively recent is due to the scarcity of medical treatment and the poor living conditions in the regions concerned, which also explains the occurrence of endemic diseases (in particular tuberculosis), the frequency of acute rheumatic fever, malaria and pyogenic infections. Prophylactic treatment by colchicine, suggested by Turkish authors and one American author, has been demonstrated to avoid not only inflammatory episodes but also the development of amyloidosis. PMID- 11915559 TI - [Hereditary intermittent fevers, other than familial Mediterranean fevers]. AB - Familial Mediterranean fever is no more the sole hereditary disease characterized by recurrent inflammatory attacks. Three other main entities have now been defined, both at clinical and genetic levels: a dominant disease due to mutations of one of the tumour necrosis factor receptor, called TRAPS for tumour necrosis factor receptor associated periodic syndrome, the hyper-immuno-globulinaemia D and periodic fever syndrome (HIDS), which is a metabolic disorder and the Muckle Wells syndrome. A thorough diagnosis of these diseases is crucial for appropriate management and treatment. PMID- 11915560 TI - [Symptomatic intermittent fever of inflammatory diseases]. AB - Intermittent fever has a wide variety of causes such as infectious, cancers, or inflammatory disease. Intermittent fever is sometimes a diagnostic challenge when fever appears as the first and isolated sign of the disease. Adult onset Still disease and juvenile chronic arthritis are mainly the most common cause of intermittent inflammatory fever. Some frequent diseases gives intermittent fever in few cases like ankylosing spondylitis, pulmonary embolism, sarcoidosis or Crohn's disease. Some rare inflammatory disease gives typical intermittent fever like genetic periodic fever. Other rare diseases give sometimes intermittent fever like vasculitis, polychondritis, Castleman disease, etc. Drug fever and factitious fever are other classical causes of intermittent fever. Diagnosis of inflammatory intermittent fever is frequently based on the clinical course but some biological tests and computerized tomographic scans are worthwhile tools. Follow-up of undiagnosed cases is needed. PMID- 11915561 TI - [Management of intermittent fever in the adult]. AB - Episodic fever is a difficult diagnostic challenge, more enigmatic than prolonged fever of unknown origin. Patients are younger, and have longer history of fever before admission. Despite the use of extensive diagnostic procedures 50% of the cases remain undiagnosed. A meticulous physical examination and history remains an important tool for registering potentially diagnostic clues useable in the diagnostic process. In the absence of clues, a staged diagnostic protocol can be used to search elements contributing to the diagnosis (laboratory investigations including chest X-ray and ultrasonography of upper abdomen, imaging techniques, endoscopies and histologic investigations. When diagnosis is not established and patient deteriorate, empiric therapeutic trial is started to presumptive diagnoses. If not, patient must be carefully followed as an out patient basis and symptomatically treated, periodic reassessment will in time reveal the development of overt disease. PMID- 11915562 TI - [Management of intermittent fever in the child]. AB - Recurrent fevers in children are common, mainly due to viral (particularly in day care centers), or to bacterial (urinary tract upper and lower respiratory) infections. The diagnosis of recurrent hereditary fever is now possible on the basis of clinical features, biochemical and genetic tests. Familial Mediterranean Fever (FMF) remains the most frequent disorder of this group, which includes now three other entities: TNF receptor associated periodic syndrome (TRAPS), the hyperIgD syndrome(HIDS) and the Muckle-Wells syndrome. PMID- 11915563 TI - [Networks of care or of health]. PMID- 11915564 TI - [Premature birth and fetal growth retardation. Risk factors and prevention]. PMID- 11915565 TI - [Acute diarrhea and dehydration in infants and children]. PMID- 11915566 TI - [Digestive hemorrhage]. PMID- 11915567 TI - [Mitral valve insufficiency]. PMID- 11915568 TI - [Prostate tumors]. PMID- 11915570 TI - [Nontraumatic spinal cord compression and cauda equina syndrome]. PMID- 11915569 TI - [Acute lymphoblastic leukemia]. PMID- 11915571 TI - The health risks of long haul flying. PMID- 11915572 TI - Compensation for asbestos-related lung cancer. PMID- 11915573 TI - Fact or fiction? Verbal and behavioural clues to detect deception. PMID- 11915574 TI - Railway disasters. PMID- 11915575 TI - Recent research into dealing with the problem of malingering. PMID- 11915576 TI - The impact of different doses of medroxyprogesterone acetate on mood symptoms in sequential hormonal therapy. AB - The aim of this study was to compare adverse mood effects of two different doses of medroxyprogesterone acetate (MPA) during postmenopausal hormone replacement therapy (HRT) in women with and without a history of premenstrual syndrome (PMS). The study was designed as a randomized double-blind cross-over study and included 36 postmenopausal women at three health care areas in northern Sweden. The women received 2 mg estradiol continuously during five 28-day cycles and 10 mg or 20 mg MPA sequentially for 12 days during each cycle. The main outcome measures were mood and physical symptoms noted on a daily rating scale. We found that physical symptoms did not differ between 10 and 20 mg MPA. Both women with a history of PMS and women without responded with more negative mood symptoms with the lower dose of MPA. In women with previous PMS the higher dose of MPA enhanced positive mood symptoms. With respect to mood and physical symptoms, the aim to lower MPA doses in HRT is unwarranted. PMID- 11915577 TI - High concentrations of CA 125 in uterine flushings: influence of cause of infertility and menstrual cycle day. AB - Uterine flushings were obtained under transvaginal ultrasonographic control from 132 women presenting for investigation and treatment of infertility. Levels of CA 125 were measured by radioimmunoassay and results expressed in relation to the total protein concentration of the same flushings. CA 125 was detected in uterine fluid at levels higher than those previously reported in peripheral blood. Uterine fluid CA 125 concentrations varied throughout the menstrual cycle, being highest in the mid-follicular phase (days 6 to 10). Uterine fluid CA 125 concentrations may reflect endometrial secretion of this protein more directly than serum levels. CA 125 concentrations did not vary according to the cause of infertility but further work in larger numbers of women is required. PMID- 11915578 TI - Twin pregnancy using recombinant gonadotropins in a woman with hypogonadotropic hypogonadism. AB - In women with hypogonadotropic hypogonadism both follicle-stimulating hormone (FSH) and luteinizing hormone (LH) are required to induce optimal follicular growth and steroidogenesis. The development of molecular genetic technology has led to the availability of recombinant FSH and LH for the induction of follicular growth and ovulation. We describe a first case of a twin pregnancy in a 36-year old patient presenting with primary hypogonadotropic amenorrhea and empty sella syndrome and treated with recombinant FSH and LH. This therapy led to the maturation of two follicles, both of which were fertilized. A twin pregnancy ensued and two normal infants were delivered. PMID- 11915579 TI - Endocrine function in HIV-infected women. AB - Infection with the human immunodeficiency virus (HIV) results in a chronic systemic illness with multi-organ involvement, severe immunosuppression and profound cachexia. It has had a major impact on women's health. Endocrine abnormalities may contribute to the clinical presentation and therefore appropriate treatment would theoretically improve the patient's condition. This pilot study was undertaken to assess the endocrine status in a group of HIV seropositive women with the view to developing recommendations for future investigations. Thirteen women were recruited from a clinic for HIV-infected patients. All women had a comprehensive general and gynecological examination. Basal endocrine status was assessed and combined pituitary testing with gonadotropin-releasing hormone, thyrotropin-releasing hormone, growth hormone releasing hormone and corticotropin-releasing hormone was performed. None of the participating women presented with gynecological complaints or had symptoms suggestive of an endocrinopathy. On questioning, seven women complained of menstrual abnormalities. Three had a body mass index of less than 20 kg/m2. Genital tract infections were common. Endocrine assessment demonstrated abnormalities of the pituitary-adrenal, pituitary-thyroid and pituitary-ovarian axes in seven women. One woman had panhypopituitarism. In six of the seven affected women CD4 counts were below 200 cells/mm3. Alterations in endocrine function were observed in seven of the women tested. While routine endocrine testing may not be indicated in all HIV-seropositive women, we should be aware of possible subtle presentations of endocrine abnormalities which may require treatment, especially in stress situations. PMID- 11915580 TI - A comparison of glyceryl trinitrate with diclofenac for the treatment of primary dysmenorrhea: an open, randomized, cross-over trial. AB - Primary dysmenorrhea is a syndrome characterized by painful uterine contractility caused by a hypersecretion of endometrial prostaglandins; non-steroidal anti inflammatory drugs are the first choice for its treatment. However, in vivo and in vitro studies have demonstrated that myometrial cells are also targets of the relaxant effects of nitric oxide (NO). The aim of the present study was to determine the efficacy of glyceryl trinitrate (GTN), an NO donor, in the resolution of primary dysmenorrhea in comparison with diclofenac (DCF). A total of 24 patients with the diagnosis of severe primary dysmenorrhea were studied during two consecutive menstrual cycles. In an open, cross-over, controlled design, patients were randomized to receive either DCF per os or GTN patches the first days of menses, when menstrual cramps became unendurable. In the subsequent cycle the other treatment was used. Patients received up to 3 doses/day of 50 mg DCF or 2.5 mg/24 h transdermal GTN for the first 3 days of the cycle, according to their needs. The participants recorded menstrual symptoms and possible side effects at different times (0, 30, 60, 120 minutes) after the first dose of medication on the first day of the cycle, with both drugs. The difference in pain intensity score (DPI) was the main outcome variable. Both treatments significantly reduced DPI by the 30th minute (GTN, -12.8 +/- 17.9; DCF, -18.9 +/- 16.6). However, DCF continued to be effective in reducing pelvic pain for two hours, whereas GTN scores remained more or less stable after 30 min and significantly higher than those for DFC (after one hour: GTN, -12.8 +/- 17.9; DFC, -18.9 +/- 16.6 and after two hours: GTN, -23.7 +/- 20.5; DFC, -59.7 +/- 17.9, p = 0.0001). Low back pain was also relieved by both drugs. Headache was significantly increased by GTN but not by DCF. Eight patients stopped using GTN because headache--attributed to its use--became intolerable. These findings indicate that GTN has a reduced efficacy and tolerability by comparison with DCF in the treatment of primary dysmenorrhea. PMID- 11915581 TI - Body fat distribution, insulin sensitivity, ovarian dysfunction and serum lipoproteins in patients with polycystic ovary syndrome. AB - Polycystic ovary syndrome (PCOS) is characterized by various endocrine and metabolic abnormalities, whose mutual associations and symptoms are still not clear. In the present study, fifteen PCOS patients and fifteen controls, matched for age and body weight, were investigated. Endocrine profiles were evaluated by the nafarelin and the adrenocorticotropin (ACTH) test. Insulin sensitivity was determined by an intravenous insulin tolerance test. Patients showed a significant predominance of abdominal adiposity [waist-to-hip ratio (WHR), 0.86 +/- 0.05 vs. 0.79 +/- 0.04] with markedly higher fasting insulin levels (+75%) and reduced insulin sensitivity (-37%). Fasting insulin, testosterone and free androgen index were positively correlated with the body mass index (BMI). In contrast, insulin sensitivity and BMI were inversely correlated in patients only. In the nafarelin test increases of 17-OH-progesterone and androstenedione were higher in patients and positively correlated with fasting insulin levels. Lipoprotein profiles showed trends towards higher triglycerides, lower HDL cholesterol and a preponderance of small, dense LDL in patients. In PCOS higher triglycerides and lower HDL cholesterol were correlated with insulin sensitivity. It is concluded that PCOS patients show metabolic abnormalities combined with a more adroid type of adiposity when compared to cyclic controls of similar BMI. PMID- 11915582 TI - Surgical or medical treatment of polycystic ovary syndrome: a cost-benefit analysis. AB - With the availability of laparoscopic ovarian cautery, there has been a resurgence in interest in the surgical treatment of clomiphene citrate-resistant polycystic ovary syndrome (PCOS). Comparison of ovulation and pregnancy rates has found no difference in success rates between ovarian cautery and gonadotropin ovulation induction for such women. We have therefore compared the cost of laparoscopic ovarian cautery with that of a typical cycle of gonadotropin ovulation induction, and also found that there is little difference. Because of the potential advantages of ovarian cautery, we recommend this surgery as the next line of treatment if clomiphene citrate fails to induce ovulation in PCOS patients, before gonadotropins are introduced. PMID- 11915583 TI - Effects of gonadotropin-releasing hormone analog treatment on skin condition. AB - The skin is a target organ of estrogens. Thus, theoretically, a hypoestrogenic state induced by gonadotropin-releasing hormone analog (GnRHa) treatment may have effects on skin condition. The aim of this study was to evaluate skin condition during GnRHa treatment. Sixteen premenopausal women undergoing GnRHa treatment for 16 weeks, as a presurgical treatment for uterine leiomyomas, were studied. Measurement of serum estradiol levels and epidermal hydration, and evaluation of subjective findings on skin condition using a questionnaire, were performed every 4 weeks during the treatment period. Serum estradiol levels were significantly suppressed at 4 weeks of treatment, and remained low afterwards. Epidermal hydration measured by corneometer did not show any significant difference at any time point examined, compared with that before treatment. No particular subjective findings relating to the skin (dryness, wrinkling, roughness, pigmentation, itching, formication, reaction to cosmetics) were reported during treatment, whereas complaints about hot flushes and sweating were notable. The results of this preliminary study support the notion that GnRHa treatment for 16 weeks is unassociated with apparent changes in skin condition. PMID- 11915584 TI - New alternative treatment in hirsutism: bicalutamide 25 mg/day. AB - The efficacy of low-dose bicalutamide (25 mg/day) in the treatment of hirsutism was investigated in this study. Hirsutism score was determined, according to a modified Ferriman-Gallwey scoring system, in 42 women with hirsutism. Each patient received 25 mg/day bicalutamide. Before therapy, multiscreen blood chemistry, hormonal analysis, and complete blood counts were performed. These parameters and hirsutism scores were repeated at 3 and 6 months during therapy. The paired Student's t-test was used to compare repeated values. Clinical improvement in the degree of hirsutism was observed in all patients by the same author. The modified Ferriman-Gallwey scores decreased from a mean of 22.0 +/- 5.1 to 8.6 +/- 3.5 (p < 0.0001). The reduction in hirsutism scores was 41.2 +/- 11.4% at 3 months and 61.6 +/- 11.1% at 6 months. In conclusion, bicalutamide at 25 mg/day is an effective drug in the treatment of patients with hirsutism. PMID- 11915585 TI - Effect of hormone replacement therapy on insulin secretion and insulin sensitivity in postmenopausal diabetic women. AB - The aim of the present study was to evaluate the effect of three different combinations of hormone replacement therapy (HRT) on insulin secretion, peripheral insulin sensitivity, serum lipid levels and parameters of oxidative stress. Seven type II diabetic women of mean age 55.4 +/- 4.7 years, who had been menopausal for an average of 5 years, were enrolled in the study. Phases of insulin secretion--first (FPIS) and second (SPIS)--and the area under the curve (AUC) for insulin secretion were studied during an intravenous glucose tolerance test (IVGTT). Insulin sensitivity was determined using the manual euglycemic hyperinsulinemic clamp technique. Three different HRT combinations were applied consecutively for 3-month periods: estradiol valerate plus cyproterone acetate (Climen); transdermal 17 beta-estradiol (System TTS 50) plus dydrogesterone (Duphaston) 10 mg daily for 10 days a month; oral 17 beta-estradiol plus dydrogesterone (Femoston) for 14 days a month. A group of nine women with normal glucose tolerance (according to World Health Organization criteria during a 75-g oral glucose tolerance test (OGTT)), of mean age 50.1 +/- 8.2 years and mean body mass index 24.60 +/- 2.01 kg/m2, were also studied, and served as a control group. Insulin secretion improved significantly after Climen: FPIS increased by 16% and SPIS by 44%. Insulin sensitivity increased by 50% after Systen TTS 50 + Duphaston; fasting hyperinsulinemia was normalized and total antioxidant capacity of the serum (TAOCS) was significantly raised (p < 0.01). Femoston led to an increase in insulin sensitivity (by 23%) and in TAOCS (p < 0.05), while fasting hyperinsulinemia remained unchanged. HRT should be prescribed in type II diabetic postmenopausal women because of its favorable effect on existing pathophysiological defects. Cyproterone acetate should be preferred in cases with a predominant beta-cell insulin secretion defect, while dydrogesterone in combination with a transdermal estrogen should be recommended in cases with leading insulin resistance. PMID- 11915586 TI - Short-term human chorionic gonadotropin-induced testosterone rise does not modify leptin levels in eugonadal men. AB - The aim of this study was to monitor serum leptin concentrations after altering the levels of testosterone, by intramuscular administration of human chorionic gonadotropin (hCG), in eugonadal men. A 7-day monitoring of hCG, testosterone and leptin levels was performed after intramuscular administration of a dose of 5000 IU hCG in these men. Thirty fertile men aged 23-38 years were studied. In addition, 30 women aged 18-34 years with normal ovulatory cycles were studied, to verify reports of sexual dimorphism in serum leptin levels. These 60 individuals were divided into four groups, according to their sex and body mass index (BMI) values. In men, blood samples were collected at 09.00, after an overnight fast, for the determination of hCG, testosterone and leptin levels, and, immediately thereafter, a dose of 5000 IU hCG was administered intramuscularly. Further blood samples were collected at 24-h intervals for a period of 7 days for determination of the same hormones. In women, blood samples were collected only once, at 09.00, after an overnight fast between the 3rd and the 6th day of the menstrual cycle, for determination of serum estradiol and leptin levels. Our results showed that the mean value of leptin in thin men and women was significantly lower than that in obese men and women, respectively. The mean value of leptin in thin women was significantly higher than that in obese men. Serum leptin concentrations decreased significantly, 168 h after short-term hCG administration. There was a significant positive correlation between BMI values and serum leptin concentrations, in both men and women. Our results support the view that hCG administration in eugonadal men does not influence serum leptin levels. Moreover, a short-term increase of serum testosterone levels, after one dose of hCG, is not sufficient to affect and modify leptin secretion mechanisms in vivo. PMID- 11915587 TI - The renin-aldosterone system and drospirenone. PMID- 11915588 TI - Effects of estrogen-progestin and raloxifene therapy on nitric oxide, prostacyclin and endothelin-1 synthesis. AB - This randomized double-blind study was conducted to investigate the effects of 17 beta-estradiol plus norethisterone acetate, and raloxifene, on nitric oxide (NO), prostacyclin (PGI2) and endothelin-1 (ET-1) serum levels in postmenopausal women. Treatment was initiated after a 28-50 day placebo period. Fourteen women were treated daily with 17 beta-estradiol 2 mg plus norethisterone acetate 1 mg (E2 + NETA), and 14 with raloxifene HCl 60 mg for a period of 6 months. Serum NO, PGI2 and ET-1 levels were estimated at baseline, after placebo, and at months 3 and 6. E2 + NETA decreased NO levels significantly, while raloxifene did not cause any appreciable change. Both regimens decreased PGI2 levels and ET-1 levels significantly. Finally, E2 + NETA and raloxifene increased the NO/ET-1 ratio by 61.4% and 81.1%, respectively. In conclusion, both regimens may exert a cardio protective effect by decreasing ET-1 levels and increasing the NO/ET-1 ratio. In contrast, both regimens had a negative influence on PGI2 levels. PMID- 11915589 TI - Forensic DNA data banks: considerations for the health sector. PMID- 11915590 TI - HIV serostatus disclosure: legal, scientific and ethical considerations. PMID- 11915591 TI - Patents & public health: international trade obligations and domestic policy development. PMID- 11915592 TI - Bridging the gaps in chronic illness care for children. PMID- 11915593 TI - Simple screen proves highly accurate in identifying children with special needs. AB - Research shows that the relatively small group of children with special health care needs accounts for more than 80% of all child-related health care costs, and there is plenty of evidence that the care of these children is a far cry from optimal. See what steps are being taken to give these children the attention they need, and check out a simple new tool designed to identify children with special health care needs. PMID- 11915594 TI - Colonoscopy benchmarks suggest less invasive tack not always most productive. AB - A new study of colonoscopy calls into question the conventional wisdom about its use versus that of sigmoidoscopy, and provides a host of crucial benchmarks for the nation's second most commonly performed outpatient procedure. PMID- 11915595 TI - Individualize seniors' drug therapy to reduce risk of osteoporosis fractures. AB - New evidence suggests hormone replacement therapy (HRT) may not be the best option for the prevention and treatment of osteoporosis after all, leaving clinicians to reconsider their recommendations to patients with or at risk of the devastating disease. PMID- 11915596 TI - Convert physician gripes into health care process changes. AB - Most health care organizations don't solicit complaints from physicians because they think they hear enough criticism already. Those that recognize physicians as the "purchasers" of health care are in a terrific position to increase provider satisfaction and improve overall operations, says a California consultant who specializes in performance improvement. PMID- 11915597 TI - The gentle art of touch. PMID- 11915598 TI - Journey into another realm. Seeking answers in energy medicine. PMID- 11915599 TI - Pharmacognosy. What nurses need to know about herbal remedies. PMID- 11915600 TI - Henry meets his match. PMID- 11915601 TI - Building confidence in spiritual care. Strategies & outcomes. PMID- 11915602 TI - Tristam Engelhardt's Christian bioethics. Interview by Susan A. Salladay. PMID- 11915603 TI - Biblical alternatives. PMID- 11915604 TI - When tears turn to joy. PMID- 11915605 TI - Night of terror. Reflections on the Taiwan earthquake. PMID- 11915606 TI - No nameless faces. PMID- 11915607 TI - A Christian psychiatric unit. The freedom to believe. PMID- 11915608 TI - When pills don't work. PMID- 11915609 TI - What is homeopathy? PMID- 11915610 TI - Making sense of complementary & alternative therapies. PMID- 11915611 TI - [Perioperative cortical stimulation of language fields under local anesthesia in preparation to excision of tumors of the dominant hemisphere]. AB - OBJECTIVES: The aim of this study is to demonstrate the accuracy of direct cortical stimulation of language areas preparatory to the removal of infiltrating tumors of dominant hemisphere. MATERIAL AND METHODS: From June 1998 to March 2000 we included in our study 15 patients, aged from 30 to 75 years, harboring gliomas (14 high grades and 1 low grade) close to language-specific cortex. All patients had slight inaugural phasic troubles. They underwent craniotomy under local anesthesia for cortical stimulation language mapping, in conjunction with electrocorticography to identify the after-discharge threshold. Stimulation mapping covered the entire macroscopically tumor involved area, extending up to 3 cm away from the margins of the lesion, without searching to identify systematically the language areas. Therefore, the lesion was removed as completely as possible, respecting a security margin of at least 1 cm from the recognized language sites. RESULTS: We identified from 1 to 6 language sites for 14 of our patients, in different locations. A radical removal was achieved for all high grade gliomas, while the low grade was only partially removed. A patient died on the 16th postoperative day from pulmonary embolism; 2 patients deteriorated from peri- or immediate postoperative complications; 9 showed a transient neurological worsening that receded by the 2nd postoperative month, while 3 did not present any postoperative aggravation. At a follow-up of 1 month to 2 years (mean 8 months), 6 patients died and the 9 survivors maintain a good life quality. CONCLUSIONS: Cortical stimulation for language mapping is an accurate technique that allows the surgical morbidity of lesions in proximity to language areas to decrease and become comparable to the surgical morbidity for lesions in non eloquent areas. PMID- 11915612 TI - [Functional recuperation after resection of gliomas infiltrating primary somatosensory fields. Study of perioperative electric stimulation]. AB - BACKGROUND: With the aim to achieve the most complete resection in cases of parietal low-grade glioma involving the primary somatosensory area (S I), the authors describe a series with resection of S I using an intraoperative brain mapping. The immediate and delayed postoperative functional results are analyzed. PATIENTS AND METHODS: Six patients were operated on for a low-grade glioma involving S I, revealed by seizures with a normal clinical examination. All patients underwent awake surgery under local anesthesia, to allow realization of an intraoperative sensorimotor brain mapping using direct electrical stimulations. RESULTS: The primary motor area (M I) and S I were detected in all procedures. A part of infiltrated S I was removed in spite of a sensitive response of this area during stimulations (sensory face in 2 cases, upper limb area in 5 cases, and lower limb area in 2 cases). All patients presented an immediate postoperative sensory deficit, with recovery in the 6 cases within 3 months. Five resections were total and one subtotal. CONCLUSIONS: In our experience, the primary somatosensory area could be resected in case of tumoral infiltration, without inducing a permanent neurological deficit. This may be due to brain plasticity mechanisms, with recruitment of loco-regional and controlateral eloquent regions, and also the learning of new compensatory strategies. Such a phenomenon can lead to perform more extensive resections in cases of infiltrative gliomas in the somatosensory region. PMID- 11915613 TI - [Adenoid cystic carcinomas invading the skull base. Apropos of 4 cases and review of the literature]. AB - Head and neck adenoid cystic carcinomas may invade the adjacent skull base by bone lysis and/or by perinervous and perivascular spread within the skull base foramina. Neurosurgical decision making is not well defined regarding the extent of intracranial tumor component removal, as neurosurgical expertise is limited for this peculiar type of tumors. The issue is to decide whether a radical supposedly locally curative surgery should be attempted, or if a large non disfigurating surgery is mandatory, keeping in mind the frequency of local recurrences and of distant metastases. Over a 13-year period, four adenoid cystic carcinomas invading the skull base were operated on at our institution: two tumors originated in the parotid gland, one in the sphenoid sinus, and one in the ethmoid sinus. Surgical removal was total in one case, subtotal in three cases. Post-operative irradiation was delivered in the four patients (two neutron irradiation, two conventional). One patient with advanced metastatic disease was submitted to chemotherapy. Three patients died from local tumor progression and distant metastases within three years after the intracranial tumor extension has been diagnosed. The patient with an ethmoid tumor is still alive seven years after surgery without any evidence of local tumor progression nor distant metastases. Surgery remains the gold standard treatment for adenoid cystic carcinomas invading the skull base. However, in our opinion a large tumor removal, without or with bone osteotomies, but without sacrifice of cranial nerves, cavernous sinus, internal carotid artery, and of the orbit allows patient survival with an acceptable comfort and absence of psychological distress due to disfigurating surgery nor surgically induced neurological functional deficit. Post-operative irradiation may sometimes stabilize locally the lesions. The place of chemotherapy has, yet, to be determined. PMID- 11915614 TI - [Medullary compression by Burkitt lymphoma. Analysis of 7 cases]. AB - OBJECTIVES: Burkitt's lymphoma is one of the possible cause of spinal cord compression. We defined the surgical approach in this rare localization of Burkitt's lymphoma. PATIENTS AND METHODS: We report 7 cases of spinal cord compression secondary to Burkitt's lymphoma treated at the Yopougon University Hospital from October 1990 to October 1994. RESULTS: Mean age of the patients was 15.4 years, range 8 to 28 years. Five of the 7 patients were children. Surgery was both a diagnostic and therapeutic emergency in this rare but serious localization of Burkitt's disease, which remains highly chemosensitive. CONCLUSION: Surgical management is warranted if Burkitt's lymphoma is revealed by spinal cord compression or if there is a primary spinal epidural localization. PMID- 11915615 TI - [Functional recuperation following lesions of the primary somatosensory fields. Study of compensatory mechanisms]. AB - During a long time, lesions of the primary somatosensory area (S I) were considered as generating severe and definitive neurological sequelae. However, recent reports described a recovery following stroke and resective surgeries (for epilepsy or tumor). On the basis of these observations, of experimentations in animals, and of studies of the (re)organization of eloquent areas in man using new methods of functional mapping, brain plasticity phenomena underlying the recovery were analyzed. They seems to implicate the recruitment of local areas (sensory redundancies within S I), regional areas (primary motor area, secondary somatosensory area, posterior parietal cortex, insula), controlateral eloquent regions, and even learning of new compensatory strategies. Such data could allow us to extend surgical indications of resection of lesions located within S I. An improvement of our knowledge of these functional reshaping phenomena (unmasking of latent networks and/or participation of eloquent homologous and/or sprouting) and then their prediction remains mandatory, with the goal to optimize the surgical preplanning in functional regions, taking account of the individual dynamic spatiotemporal cortical organization. This better knowledge seems to be obtained by studying the correlations between pre- and post-operative functional neuroimaging (fMRI, MEG) data, and the intraoperative functional mapping results. PMID- 11915616 TI - [Burkitt lymphoma of the cavernous sinus. Apropos of a case]. AB - Cavernous sinus lymphoma is a rare occurrence. We report a case of a 37-year-old woman who was admitted for an oculo-motor paresis and retro-orbital pain. CT scan and MRI disclosed a cavernous sinus tumor. A surgical biopsy was performed through a pterional approach and the histological diagnosis was a solitary primary Burkitt's lymphoma. Aggressive chemotherapy led to a complete and asymptomatic remission after a follow-up of 9 years. The mechanism of the primary localization in the cavernous sinus is discussed. PMID- 11915617 TI - [Tuberculous abscess of the cerebellum. Apropos of a case]. AB - A 22-year-old Asian man developed intracranial hypertension with a 38 degrees C fever. Two months earlier, he had undergone surgery and medical treatment for tuberculous otomastoiditis and pulmonary tuberculosis. The CT scan revealed multiple tuberculous abscesses of the cerebellum. Histological, microbiological and biological proof of diagnosis was obtained. We advocate surgical treatment of intracranial tuberculous abscesses associated with antituberculosis chemotherapy for 18 months. The earlier the treatment, the better the prognosis. This pathology must be kept in mind when treating patients from countries with a high endemic rate of tuberculosis and suffering from immunodeficiency syndrome. PMID- 11915618 TI - [Ossification of ligamentum flavum unmasked by acute paraplegia]. AB - A 30-year-old black man presented sudden-onset paraplegia during a foot-ball match, after a movement of hyperextension of the trunk. Moreover, the patient exhibited an hypoesthesia below the T11 level, with sphincter disturbances. The MRI and the CT-scan showed a stenosis of the spinal canal related to an ossification of hypertrophied ligamenta flava from T10 to T12. Intramedullary abnormal signals on MRI images were compatible with a spinal cord hemorrhage. A laminectomy with removal of abnormal ligamenta flava was carried out, and their endochondral ossification was confirmed by pathological examination. Two months later, the patient was able to walk alone and exhibited a mild spasticity associated to sensory disturbances of lower limbs. Ossification of ligamenta flava is usually observed in Japanese patients, sometimes in Caucasians, more rarely in black people. Its mechanism is unclear except when associated with metabolic or endocrine diseases. The patients usually present with clinical features of chronic spinal cord compression. Our case seems to be the first one disclosed by an acute spinal cord injury on ossified ligamenta flava. In this patient, because of remaining adjacent ossified ligamenta flava and the development on postoperative MRI of an intramedullary cavity, a long-term clinical and radiological follow-up is particularly necessary. PMID- 11915619 TI - [Potential neurosurgical damage of rubber bullets. Analysis of 2 pediatric cases]. AB - We present 2 cases of craniocerebral injuries in children caused by rubber bullets. The potential severity of these projectiles is discussed. The first case was an eleven-year-old boy who died a few days after having been injured by a rubber bullet during a riot. He presented with a bi-hemispheric fronto-parietal craniocerebral injury with massive intraventricular hemorrhage. The second case was a fourteen-year-old boy also injured by a similar rubber bullet with an important left fronto-temporal cerebral contusion. These projectiles were composed of a metal cylinder (length: 1.7 cm, diameter: 1.7 cm) covered by a 2 mm layer of rubber. The term "rubber bullet" could give the impression that these projectiles are harmless. But, based on these observations and on the literature, these rubber bullets (like their predecessors) can induce severe, or even fatal craniocerebral injuries. PMID- 11915620 TI - Modification of electrochemically deposited apatite using supercritical water. AB - Supercritical water was used as a modification method of electrochemically deposited apatite on pure titanium. The apatites were coated on a commercially pure titanium plate using a hydrothermal-electrochemical method. A constant direct current at 12.5 mA/cm2 was loaded for 1 hr at 25, 60, 100, 150 and 200 degrees C in an electrolyte containing calcium and phosphate ions. The deposited apatite on the titanium substrate was stored in supercritical water at 450 degrees C under 45 MPa for 8 hr. With this treatment, the crystallinity of the apatites increased, sharp edges of the deposited apatites were rounded off, and the bonding strength of the titanium substrate to the deposited apatites significantly increased. On the other hand, weight loss in 0.01 N HCl decreased and the weight gain rate in a simulated body fluid also decreased with this treatment. It is suggested that the modification using supercritical water improved the mechanical strength of the deposited apatite, but worsened its bioactivity. PMID- 11915621 TI - Fabrication of a functionally graded dental composite resin post and core by laser lithography and finite element analysis of its stress relaxation effect on tooth root. AB - Laser lithography was applied for Computer Aided Design and Computer Aided Manufacturing (CAD/CAM) fabrication of dental prostheses made of composite resin. First, the conditions to obtain the optimum resolution for photo-curing were determined, and then a composite resin full crown was fabricated by laser lithography. Second, a functionally graded composite resin post and core which had gradient elasticity in the post, was manufactured by the gradual change in the filler contents of the composite resin. Finally, stress analysis of the functionally graded post and core was performed by a two-dimensional finite element method. This demonstrated the effects of reducing the stress concentration around the apex of the post. PMID- 11915622 TI - Effect of experimental fluoride-releasing tooth separator on acid resistance of human enamel in vitro. AB - This study aimed to investigate the fluoride-releasing ability of an experimental tooth separator consisting of polyurethane elastomer with tin fluoride and its effect on the acid resistance of human enamel. The tooth separator was set around an enamel slab and stored in de-ionized water for 10 days. The daily concentration of fluoride in the de-ionized water was measured. Then the enamel surface was artificially decalcified by a lactic acid buffer solution (pH 4.5) for 96 hours. The mineral density at the surface layer of the enamel was measured to evaluate the acid resistance. The fluoride release increased with the amount of fluoride in the separator, but decreased with the immersion time. Both the enamel area contacting with the separator and its surrounding area showed lower mineral loss and lesion depth compared with the controls (P < 0.05). It is suggested that the experimental tooth separator would release enough fluoride and improve the acid resistance of the enamel surface layer. PMID- 11915623 TI - Effects of Sn, Ga, and In additives on properties of Ag-Pd-Au-Cu alloy for ultra low fusing ceramics. AB - Nine 35% Ag-30% Pd-20% Au-15% Cu alloys containing 2, 4 and 6 mass% of Sn, Ga or In as an additive metal were experimentally prepared to investigate the effects of different additives and their content on the physical and mechanical properties as well as the bond with a ultra-low fusing ceramic. Both the different additives and their content or either of these two factors significantly influenced most of the evaluated properties except for the area fraction of the retained ceramic. Based on the evaluated properties three experimental alloys (2% Sn-added alloy, 4% Ga-added alloy and 2% In-added alloy) can be recommended as a suitable alloy for ceramic-metal restorations using ultra low fusing ceramics. PMID- 11915624 TI - Corrosion behavior of pure titanium and titanium alloys in fluoride-containing solutions. AB - The effects of fluoride concentrations and pH on the corrosion behavior of pure titanium, Ti-6Al-4V, Ti-6Al-7Nb alloys and a new Ti alloy adding palladium, which is expected to promote a repassivation of Ti were examined by anodic polarization and corrosion potential measurements. The amount of dissolved Ti was analyzed by inductively coupled plasma mass spectroscopy. The surface of the specimen was analyzed by X-ray photoelectron spectroscopy before and after the measurement. Pure Ti, Ti-6Al-4V and Ti-6Al-7Nb alloys were easily corroded even in a low fluoride concentration in an acidic environment. The corrosion resistance of Ti 0.2Pd alloy was greater than those of pure Ti, Ti-6Al-4V and Ti-6Al-7Nb alloys in the wide range of pH and fluoride concentrations. The high corrosion resistance of Ti-0.2Pd alloy was caused by the surface enrichment of Pd promoting a repassivation of Ti. The Ti-0.2Pd alloy is expected to be useful as a new Ti alloy with high corrosion resistance in dental use. PMID- 11915625 TI - Fracture aspects of resin-dentin bonding in non-trimming microtensile test. AB - Comparative studies on resin-dentin bond strength and failure mode were performed between the conventional tensile test and the microtensile test with non-trimming small specimens, 1 x 1 mm in cross-section, for two brands of dentin bonding systems. The fracture surface of the conventional large specimen showed a catastrophic cohesive failure in dentin at its center and a lesser adhesive failure, suggesting that the whole failure was due to the development of some major cracks. The non-trimming microtensile test showed significantly larger average bond strength with markedly larger standard deviation and significantly larger fraction of adhesive failure than the conventional test. Some small specimens were extremely strong and some were weak according to the heterogeneous distribution of tight bonding and defective or deficient bonding over the whole dentin surface. These results suggest that the non-trimming microtensile test may potentially provide more realistic aspects of resin-dentin bonding than the conventional bulk specimen. PMID- 11915627 TI - Experimental ammonia-free phosphate-bonded investments using Mg(H2PO4)2. AB - In previous study, we found that Mg(H2PO4)2 instead of NH4H2PO4 was available as a binder material for phosphate-bonded investments and possibly could be used to develop the phosphate-bonded investment without ammonia gas release. The purpose of the present study was to develop the experimental ammonia-free phosphate bonded investments by investigating suitable refractories. Mg(H2PO4)2.nH2O and MgO were prepared as a binder. Cristobalite and quartz were selected as refractories. The power ratio of MgO/Mg(H2PO4)2.nH2O was set constant at 1.2 according to our previous findings. Fundamental properties of dental investment such as strength, manipulation and expansion were evaluated. Using cristobalite as the refractory material, further investigations were performed. The refractory/binder ratio was definitely effective. The increase of this ratio led to low mold strength and large mold expansion. The present findings suggested that C5 was desirable for dental investment. PMID- 11915626 TI - Radiation effects of carbon ions and gamma ray on UDMA based dental resin. AB - The radiation effects on the mechanical and physical properties of photo polymerized UDMA resin without filler was investigated by various mechanical tests and spectroscopic measurements. The radiation sources were carbon ion (12C ion) and gamma ray (gamma-ray). With 640 Gy of 12C ion radiation, Vickers hardness increased by about 40%, the degree of abrasion decreased by 30%, and the flexural strength increased by 20%. With the same dose of gamma-ray radiation, only Vickers hardness increased by 19%. The spectra taken by the Fourier Transform Infrared Spectrometer (FT-IR), Raman spectroscopy, and Fluorescence spectrophotometer showed little change in the peak configuration and background intensities. The relative degree of conversion (DC) of carbon double bonds by radiation to the state of non-radiated samples were estimated by FT-IR. Spectroscopic results were indicative for the formation of cross-linking between carbon chains. Cross-linking of carbon molecules induced by radiation might be one of the reasons for the improved mechanical properties of UDMA resin. PMID- 11915628 TI - Characterization of powder components of commercial bone cements. AB - Acrylic bone cements have been used in orthopedic surgery without detailed information on their basic characteristics, especially on their powder components. In this study, the powder components of seven bone cements available on the market in Japan were characterized for morphology, polymer structure and molecular weight, content of residual monomer and benzoyl peroxide (BPO), and thermal properties using scanning electron microscopy, nuclear magnetic resonance spectroscopy, size exclusion chromatography, high performance liquid chromatography, and differential scanning calorimetry, respectively. Considerable differences between the seven bone cements were found in polymer structure and molecular weight, and especially in BPO content and in the morphology of the polymer particles such as shape, size and distribution. It was found that the BPO content was not always in agreement with the value given by the manufacturers on the package. PMID- 11915629 TI - Change in the bisphenol A content in a polycarbonate orthodontic bracket and its leaching characteristics in water. AB - The change in the bisphenol A (BPA) content in a polycarbonate (PC) orthodontic bracket and its leaching characteristics were studied by immersing the bracket in water because BPA has been the matter of controversy in dentistry due to its estrogenic potential. PC brackets were placed in water at 37 degrees C and 60 degrees C and the BPA content in the bracket and the amount of BPA released into the water were analyzed at appropriate intervals by high performance liquid chromatography. The BPA content increased in the water with time and was 3.8-fold after 12 months at 37 degrees C and 12.4-fold after 14 weeks at 60 degrees C compared with the virgin value. The rate of BPA release also increased with time. The results suggested that BPA was released from the bracket time-dependently in the oral cavity. However, it was assessed that little or no estrogenic effect due to the released BPA is expected in the human body. PMID- 11915631 TI - Taking advantage of the information age. PMID- 11915630 TI - Efficacy of dentin bonding to cervical defects. AB - The bonding efficacy of sclerotic dentin was determined by measuring the polymerization contraction gap width of a commercial light-activated resin composite in a cervical defect and by measuring the micro-Vicker's hardness at the dentin adhesive surface; morphological characteristics were observed using a scanning electron microscope in extracted human incisors and premolars. Contraction gap formation was completely prevented when the cavity wall was primed with 35 vol% glyceryl mono-methacrylate solution after 0.5 mol/L EDTA conditioning. The contraction gap width was significantly decreased when the resin composite was filled into the sclerotic dentin cavity even when priming was omitted. These results suggest that the sclerotic dentin, which is frequently observed in cervical defects, should be preserved as a substrate because it exhibits an effect of dentin priming and is suitable for bonding. PMID- 11915632 TI - Site development in periodontal therapy--alveolar bone augmentation as an adjunct to endosseous implant placement. AB - This article reviews some of the current concepts of site development in periodontal therapy as well as hard tissue augmentation as an adjunct to endosseous implant placement. This article reviews the historical perspectives of guided bone regeneration and addresses the management and development of extraction sites, inadequate ridge widths, and sinus augmentation before implant placement. Selected cases are presented to illustrate these clinical situations. In addition, key concepts for the success of bone augmentation and implant dentistry are discussed. PMID- 11915633 TI - Caries detection using laser fluorescence. PMID- 11915634 TI - A pilot clinical evaluation of a new injection denture base system. AB - The purpose of this pilot study was to determine if clinically acceptable complete denture prostheses can be consistently fabricated with a new injection system. In the first part of this investigation, 14 patients requiring complete denture fabrication had one prosthesis made with the new injection system and one made with the compression-pack technique. Polish, porosity, and esthetics were evaluated by one of three investigators at the placement visit. The percentage of the intaglio surface in contact with the mucosa before adjustment was also measured. One week after placement, the patients were questioned regarding the prostheses' stability, the presence of any undesirable taste, and overall satisfaction. The second portion of the study involved materials property testing of flexural strength and Izod toughness of conventionally packed and injection packed specimens processed by the fast cycle or the long cycle. In every case, acceptable results were obtained for the resin's polishability, its lack of porosity, esthetics, stability, lack of unpleasant taste, and overall patient satisfaction with the prostheses. No difference in clinical adaptation was observed. These results indicated that no clinical difference was detectable between the conventionally packed and injection-packed denture bases by the investigators or patients in this study. Physical property testing indicates that the differences between groups may be more affected by the polymerization cycle used than the way that the resin was placed in the mold space. PMID- 11915635 TI - Juvenile dermatomyositis: literature review and report of a case. AB - Dermatomyositis is a rare disease of unknown origin, which affects both children and adults. In the juvenile form, dermatomyositis is a multisystem disease, characterized by myositis; an erythematous rash over the bridge of the nose, around the eyes, and on the trunk and limbs; vasculitis; and dilatation of the capillaries in the nail beds and gum margin. Late development of calcinosis is seen in approximately two-thirds of the patients. This case describes a 5-year old dental patient diagnosed with dermatomyositis. This article includes the clinical manifestations of dermatomyositis in the body and in the oral cavity as well as the considerations that should be taken when treating these patients. PMID- 11915637 TI - Composite resin abutment carriers for implant-retained prostheses. PMID- 11915636 TI - Immediate loading of dental implants in the edentulous mandible: a preliminary case report from an international prospective multicenter study. AB - The ability to predictably achieve long-term osseointegration in patients with compromised anatomical resources has been demonstrated numerous times in modern oral implantology. Recently, clinical attention has focused on new methods of reducing treatment time. One-stage surgical procedures and immediate loading of implants at the time of placement are two techniques that have demonstrated promising clinical results. A prospective clinical study of immediately splinting and loading a new, one-stage implant is currently in progress in the United States and France. An overview of the implant design and presentation of one case study from the University of Pittsburgh demonstrates how this promising technique is performed. PMID- 11915638 TI - Evaluating the efficacy of 2-octyl cyanoacrylate bioadhesive for treatment of oral ulcerations. AB - Studies were recently completed that evaluated a novel bioadhesive device for use in treating oral ulcerations. This article reviews the research protocols and results of these studies on the bioadhesive's safety, ability to decrease immediate and long-term pain, effects on ulcer healing, and equivalence to a predicate device in providing pain relief for oral ulcers. PMID- 11915640 TI - Recurrent oral ulcers--an overview. AB - Recurrent oral ulcers (ROUs) are the most common oral mucosal disease. The etiology of ROUs is complex. The factors include mechanical trauma, genetics, stress, smoking, and viral and bacterial infections. Treatment modalities depend on the differential diagnosis of ROUs and could consist of antimicrobial agents, anti-inflammatory agents, immunomodulators, or over-the-counter medications. New therapy available in the form of a coating polymer, Colgate ORABASE Soothe.N.Seal, is clinically proven to provide rapid relief and healing of ROUs. PMID- 11915639 TI - Clinical treatment evaluations of a new topical oral medication. AB - Clinical symptoms of viral infections, such as herpes labialis (cold sores) and oral ulcers (canker sores), are common in adulthood, affecting approximately 19% of adults aged 25 to 44 years. For some patients, lesions last longer, are more painful, and are more difficult to treat, requiring medications that allow the affected area to heal but do little to alleviate pain. Conventionally, pain caused by oral ulcerations has been addressed with either topical analgesics or systemic pain medications. However, a recently introduced over-the-counter topical medication (ORABASE Soothe.N.Seal) has been shown to be an effective pain relieving adjunct to healing therapies. This article reviews the characteristics of painful oral ulcerations and details the research findings of a clinical trial of the use of this new medication. PMID- 11915641 TI - Cyanoacrylate medical adhesives--a new era Colgate ORABASE Soothe.N.Seal Liquid Protectant for canker sore relief. AB - Cyanoacrylate, a synthetic adhesive, is a fast polymerizable liquid monomer. Serendipity led to the discovery of cyanoacrylate adhesives in 1951. Today, a specific cyanoacrylate monomer, 2-octyl cyanoacrylate, is being used in a topical medical adhesive formulation. The only over-the-counter cyanoacrylate-based product cleared by the Food and Drug Administration is Colgate ORABASE Soothe.N. Seal Liquid Protectant. Upon application, this liquid monomer formulation polymerizes instantly into a thin, flexible polymer film that adheres tenaciously to mucosal tissue. This polymer film creates a mechanical barrier that provides immediate and long-term pain relief of oral ulcerations and irritations, and maintains a natural healing environment for the area to heal. PMID- 11915642 TI - Magnification and the alternatives for microdentistry. AB - The evolution of microdental treatments has necessitated that dentists improve the visual acuity of the operating field in an effort to diagnose disease earlier and to treat those areas with greater precision. Alternatives in magnification exist for all practitioners from introductory magnification systems, such as single-lens loupes, to high-powered dental operating microscopes. An opportunity exists for every practitioner to enhance their vision through magnification and illumination. PMID- 11915643 TI - Understanding the applications of microdentistry. AB - Microdentistry is a science-based tool of operative dentistry, and is an alternative to traditional dentistry. The structures and substructures of teeth have been generally misunderstood. Once these structures are understood, the goal of the microdentist is to identify unsound tooth structure, which is a threat to the long-term stability of the tooth, and then neutralize or remove those structures with minimal disruption of the surrounding sound tooth structure. Minimal disruption of sound tooth structure provides the additional patient benefit of reducing or eliminating the pain associated with traditional dentistry. This article reviews some of the newly discovered structures in human teeth and discusses methods of microdentistry used to preserve these structures. PMID- 11915645 TI - Air abrasion in an era of declining caries. AB - As with all therapeutic and diagnostic technologies (both new and old), diagnostic tools must be applied in the context of the diseases they affect. This article examines the evidence for the use of air abrasion as a diagnostic tool in a time of declining dental caries. This examination is accomplished by first being clear about what evidence should be acceptable to clinicians to make any diagnostic or therapeutic changes in their practice. It then examines the evidence as it relates to air abrasion and the diagnosis of dental caries. PMID- 11915644 TI - The use of air abrasion and glass ionomer cements in microdentistry. AB - Microdentistry principles are based on the early control of and intervention in the effects of the caries process. Once a decision to treat a lesion has been made, a further understanding of the biomechanical functioning of the tooth in relation to anatomical structures is required. Preservation of these structures requires the development of skills to allow the preparation of minimally invasive cavity designs. This may involve learning to use magnification, air abrasion, caries detection dye, or a diagnostic laser. Autocure glass ionomer cements complement minimally invasive cavity designs. The preservation of healthy tooth structure using noninvasive treatment techniques or minimally invasive cavity designs is now one of the primary goals of modern operative dentistry. PMID- 11915646 TI - The role of pit-and-fissure discoloration in caries assessment. AB - Occlusal dental caries often is not apparent using traditional diagnostic techniques. The clinical significance of pit-and-fissure discoloration in the absence of dietary and habit substances was examined. The study included 462 extracted teeth (216 with obvious pit-and-fissure discoloration, 216 with subtle discoloration, and 30 with no discoloration); 130 clinical teeth with varying degrees of pit-and-fissure discoloration; and 159 teeth in young adults, which had sealants placed > or = 10 years. All teeth studied were excavated very conservatively using air abrasion and/or the uniquely small H1 004 carbide bur. Presence of caries and its depth and extent were recorded, photographed, and measured with a custom made calibrated probe. Of the 721 teeth with discolored pits and fissures studied (432 extracted teeth and 289 clinical teeth), 660 (92%) had two or more of the four clinical criteria used to define dental caries in this study. Sixty-four percent of the lesions were > 2 mm in depth and 27% were > 3 mm in depth. Of the 159 teeth sealed for > or = 10 years, 47 (92%) were carious, and 26% had large, deep carious lesions penetrating > 3 mm. These data indicate: more effective methods are needed to diagnose pit-and-fissure caries and presence of pit-and-fissure discoloration in the absence of substances causing extrinsic staining should be a strong warning for clinicians to examine carefully for dental caries. PMID- 11915647 TI - Evolution of dental ceramics. AB - A porcelain system has been developed that combines dental-ceramic engineering and fiber-optic technology. This system involves cutting-edge technology that results in optical properties that emulate nature closer than ever before. The enhanced opalescence and fluorescence, combined with optical clarity, results in color-reactive porcelain that looks natural and esthetic in any light, and reacts to light in the same manner as the natural dentition. PMID- 11915648 TI - Have impression materials made the clinician better? PMID- 11915649 TI - Can a denture be esthetic dentistry? PMID- 11915650 TI - Dentin bonding: what is the state of the art? AB - Currently, two approaches are available for bonding resin-based materials to dentin--total-etch and self-etch. The total-etch approach, using either a three step or a two-step method, has been available for several years. Clinical studies have demonstrated excellent success rates for total-etch adhesives, but many clinicians report problems with postoperative sensitivity. The self-etch approach, which is relatively new to the US market, reduces the incidence of postoperative sensitivity. However, the long-term clinical performance of self etch materials--particularly those that use a single solution to etch, prime, and bond--is not yet proven. This article provides an overview of the current options for bonding resin to dentin. PMID- 11915651 TI - 4-META as an acidic monomer in an all-in-one experimental adhesive. AB - This article describes a clinical study of an experimental, all-in-one, self etching priming enamel and dentin adhesive. This new compound is a hydroxyethylmethacrylate (HEMA)-free resin mixture of urethane dimethacrylate (UDMA) and 4-methacryloxyethyl trimellitate anhydride (4-META) dissolved in acetone and water. The pH of the adhesive solution is 2.2, which classifies this material as moderately strong. Bond strengths to enamel and dentin are approximately 20 MPa. The marginal inspection of bonded resin composite restorations in 3.5-mm wide, cylindrical, butt-joint dentin cavities reveals gap free intact margins in 6 of 8 cases. Scanning electron microscopy analysis of the bonding interface to enamel shows a shallow microretentive etching pattern, and dentin is hybridized to a depth of approximately 1 micron. The fracture morphology analyzed on debonded shear strength specimens illustrates cohesive failure in resin close to the enamel and dentin bonding interfaces. PMID- 11915653 TI - New directions for posts in restoring endodontically treated teeth. PMID- 11915652 TI - Nociceptive trigeminal inhibition--tension suppression system: a method of preventing migraine and tension headaches. AB - Dentists and physicians see countless patients suffering from various types of headaches. Various modes of therapy are used in an attempt to treat these patients. As a result of this study, it appears that a common factor to migraine and tension-type headaches may be chronic clenching. If this is the case, then dentists may be able to treat headache patients more effectively than previously suspected through the use of a dental appliance. PMID- 11915654 TI - Evaluating a dental patient for local anesthesia allergy. AB - The determination of particular allergens with respect to local anesthetics may present difficulty within the realm of clinical dentistry. A case is presented that dramatizes the difficulties in determining the specific allergen after a patient undergoing several episodes of restorative dentistry with several varying regimens of local anesthesia repeatedly reacted with skin rashes and pruritus approximately 36 hours after treatment. The patient was a 76-year-old man with a complex medical history. A challenge procedure performed with a commercial formulation of local anesthesia resulted in a positive delayed hypersensitivity reaction. An additional challenge procedure with cardiac lidocaine resulted in a negative challenge. A latex-induced delayed-type hypersensitivity reaction was suspected but unproven because the patient declined further allergy testing. Therefore, the conclusion was that the allergen was an unknown substance within the commercial local anesthetic formulation. PMID- 11915655 TI - Processed composite resin--a versatile restorative material. PMID- 11915656 TI - The functionally fixed restoration: a third modality of treatment. AB - Most dental practitioners as well as their patients prefer to have fixed rather than removable restorations. However, there are many clinical situations that prohibit the use of the fixed modality. These clinical cases can vary from simply not having the proper number of healthy teeth and/or implants to the esthetically challenging cases of high smile lines and severe loss of alveolar support. The approach of using a traditional removable prosthesis in these situations has always been met with severe compromises. The functionally fixed restoration is a third modality of treatment that can solve many of the problems of the removable restoration and at the same time provide the same comfort and success of the fixed prosthesis. PMID- 11915657 TI - Alternative implant impression techniques. PMID- 11915658 TI - Incorporating locally delivered periodontal treatment into your practice. PMID- 11915659 TI - Take control of your ceramic restorations in 2002. PMID- 11915660 TI - [Tiedemann's useful idiots]. PMID- 11915661 TI - [From waiting lists to patient shortage?]. PMID- 11915662 TI - [The epicrisis in an extended consumer perspective]. PMID- 11915663 TI - [Intensive chemotherapy in Burkitt's lymphoma and aggressive non-Hodgkin's lymphoma]. AB - BACKGROUND: Clinical studies over the last 20 years using more intensive cytostatic regimens show improved results in children and adolescents with aggressive non-Hodgkin's lymphoma and in adult patients specifically with Burkitt's lymphoma. MATERIAL AND METHODS: We present a retrospective analysis of the use of the Berlin-Frankfurt-Munster (BFM) regimen for patients older than 15 years from three Norwegian university hospitals during the 1992-99 period. RESULTS: Survival data for 24 patients 15-69 years old with Burkitt's lymphoma/B cell acute lymphoblastic leukaemia (B-ALL) show an estimated overall five year survival of 70% (75% for Burkitt's lymphoma only). Eight of ten adolescent patients 15-20 years old with other aggressive lymphomas were alive and disease free at last follow-up. All nine patients given the regimen after failure of prior therapy died of lymphoma within six years. INTERPRETATION: The BFM regimen yields impressive results as the primary treatment of adolescent and adult patients with Burkitt's lymphomas/B-ALL. PMID- 11915664 TI - [Use of the B-Lynch suture technique in postpartum hemorrhage]. AB - BACKGROUND: Serious postpartum haemorrhage occurs in approximately 1.4% of all deliveries in Norway and may result in hysterectomy. Stepwise devascularisation and hysterectomy are technically difficult and time consuming. A simple suture technique, B-Lynch suture, has recently been introduced, by which atonic postpartum haemorrhage may be controlled and hysterectomy avoided. MATERIAL, METHODS AND RESULTS: We have used the B-Lynch technique in five patients with serious postpartum uterine atonia and haemorrhage. In all patients serious bleeding and hysterectomy were avoided. Three of the patients are described in detail, one with failed induction of labour resulting in Caesarean section, one with placenta previa, and one with prolonged normal vaginal delivery. INTERPRETATION: If performed early, the B-Lynch suture technique is less mutilating to the woman; it may prevent excessive blood loss and is probably beneficial for the patient's future fertility. It is easy to perform in Caesarean sections and can be adopted in vaginal deliveries. PMID- 11915665 TI - [Paroxysmal nocturnal hemoglobinuria--a rare disease with many faces]. AB - BACKGROUND: Paroxysmal nocturnal haemoglobinuria (PNH) is a rare acquired clonal disorder of haematopoietic stem cells causing intravascular haemolysis, haemoglobinuria and occasionally severe venous thrombotic complications. PNH is associated with severe aplastic anaemia. MATERIAL AND METHODS: A total of nine patients referred to and followed at Rikshospitalet University Hospital over a period of more than 20 years were evaluated and compared with other patient materials. RESULTS: All patients are alive with a median observation time of 46 months (7-258 months). One patient has only limited clinical signs of PNH six years after diagnosis, one patient underwent allogenic bone marrow transplantation with an HLA-identical sibling and is healthy and free of disease nine and a half years posttransplant. Three patients have experienced severe thrombotic episodes, two with development of Budd-Chiari's syndrome despite anticoagulant treatment. Five patients including these two patients are being treated with anticoagulant therapy. Two patients probably having PNH have gone through pregnancies with normal delivery of healthy children. INTERPRETATION: These nine cases illustrate the many different aspects of this rare disease, including a relatively favorable prognosis with respect to survival. PMID- 11915667 TI - [Molecular genetic diagnosis of Charcot-Marie-Tooth disease and hereditary neuropathy with liability to pressure palsies]. AB - Charcot-Marie-Tooth (CMT) disease is the most commonly inherited disorder of the peripheral nervous system; in Norway, the estimated prevalence is approximately one in 2,500. CMT has been classified into a demyelinating form (CMT1) and an axonal form (CMT2). Around 70-80% of CMT1 cases are caused by a dominantly inherited 1.5 Mb duplication at 17p11.2-12 (CMT1A), encompassing the peripheral myelin protein 22 (PMP22) gene. In contrast, hereditary neuropathy with liability to pressure palsies (HNPP) is caused by the reciprocal deletion of the same 1.5 Mb region. We recently developed a method by real-time quantitative polymerase chain reaction (PCR) for detecting CMT1A duplication and HNPP deletion. Real-time quantitative PCR is very sensitive for identifying PMP22 gene copy number in CMT1A duplication and HNPP deletion. We discuss molecular genetic testing of CMT1A duplication and HNPP deletion. Real-time quantitative PCR should find broad application in clinical and research settings, not only in CMT1A duplication and HNPP deletion, but also in other diseases involving gene copy number or gene expression. PMID- 11915666 TI - [Dopa-responsive dystonia--a hereditary dystonia easy to treat]. AB - BACKGROUND: Dopa-responsive dystonia is a genetically determined disorder with early onset. The dystonia usually manifests as a disturbance of gait with fatigue and may be confused with spasticity. The diagnosis is based on clinical recognition and response to l-dopa, which is usually complete and long lasting. The most common genetic defect involves the gene for GTP cyclohydroxylase I. MATERIAL AND METHODS: We describe a Norwegian family in which three generations are affected. RESULTS: All those affected had gait disturbance from childhood; the disturbance became worse during the day and after exercise. Clinical examination revealed reduced fine motor skills and brisk tendon reflexes. Dystonic posturing of one or both legs could be seen during walking. All patients were treated with l-dopa with excellent effect. INTERPRETATION: Though uncommon, this disorder is an important differential diagnosis in children with gait disturbance, particularly in those suspected as having spastic paraparesis. PMID- 11915668 TI - [Patient referral and patient volume in a regional hospital]. AB - BACKGROUND: A general trend in health care is the increasing discrepancy between resources available and the volume of patients treated. Few studies have looked at a reduction in patient referral as a possible explanation for this situation. MATERIAL AND METHODS: The volume of hospital admissions and outpatient care 1996 through 2000 at a large regional hospital was related to the number of referrals over the same period. RESULTS: There was an overall increase in patient admissions as well as in outpatient care during the study period. However, while the number of new referrals seen as outpatients dropped by 1.2%, the number of patients in for controls increased by 27.1%. While some departments had an increase in both admissions and new outpatients, other departments had a dramatic decrease in new patient contacts but total patient volume was maintained by a comparable increase in control consultations. At the same time, there was a 15.1% overall drop in new patient referrals (new referrals for admissions down 44.4%, new referrals for outpatient care down 11.4%), in some departments even more. INTERPRETATION: The study shows that patient demand for hospital care is not unlimited and should also be considered when more resources allocated to hospitals do not result in increased patient volume. PMID- 11915669 TI - [Should patients automatically get copies of their medical reports?]. AB - BACKGROUND: Since March 1997, the Department of Neurosurgery at Tromso University Hospital has sent a copy of their medical report to all in-patients after discharge. MATERIAL AND METHODS: We performed telephone interviews with 100 patients in order to obtain their views on getting a copy of the medical report after discharge. RESULTS: All patients were satisfied with getting the report. They found the reports to be informative and a good supplement to the verbal information given at discharge. No-one, not even the seriously ill patients, had been frightened by the medical information in the reports. INTERPRETATION: Sending a copy of the medical report to all in-patients after discharge increases confidence between patient and doctor and improves patient satisfaction. PMID- 11915670 TI - [Ethical aspects of xenotransplantation]. AB - The motivation for research on animals as a source of cells, tissues and organs for human transplantation (xenotransplantation) is the scarcity of suitable donors for patients with end-stage organ failure. Use of animals in both experimental and clinical settings raise considerable ethical concerns, especially when non-human primates are used. The pig may represent an acceptable source animal for xenotransplantation and is today regarded as the animal of choice. Xenotransplantation seems defensible from an ethical perspective, but should not be performed in humans until preclinical animal-to-animal xenotransplantations have a predictable rate of success and the risk to society at large is negligible. For the individual patient and his or her next of kin involved, issues of autonomy and proper consent procedures must be given high priority. Safeguards must be established, for the patient and for society at large, in relation to health hazards from infectious organisms, particularly retroviruses. Public monitoring of research on and clinical use of xenotransplantation is mandatory. PMID- 11915671 TI - [Drug therapy of irritable bowel syndrome]. PMID- 11915672 TI - [BE smokeFREE--evaluation of an intervention program for smoke-free schools]. AB - BACKGROUND: We present an evaluation of school-based intervention aiming at smoke free schools (BE smokeFREE). The intervention programme was developed by the Norwegian Cancer Society. MATERIAL AND METHODS: A large-scale field experiment (initial n = 4,215) was carried out among lower secondary school students, starting in 8th (age 15) grade during the school year 1994/95. Schools were allocated to four groups: Control (A), a model intervention which included involvement of parents as well as courses for teachers (B), intervention identical to B but with no courses for teachers (C), and intervention identical to B but with no involvement of parents (D). Questionnaire surveys were administered four times (baseline and three follow-up surveys). A total of 19 school hours were used for the intervention, which took place in grades 8, 9, and 10. RESULTS: Changes in smoking frequency (daily, weekly, more seldom, non smoker) and number of cigarettes smoked per week were used as end-point measures. Changes in smoking habits were markedly more favourable among students in model intervention schools than among students in control schools. INTERPRETATION: Carefully planned school-based intervention contributes to reduced smoking among adolescents. PMID- 11915673 TI - [BE smokeFREE--from trials to national implementation]. AB - BACKGROUND: After small-scale trials, BE smokeFREE, a tobacco education programme covering the three years of lower secondary school was offered to all Norwegian schools in 1997. We have studied the effects of the programme, comparing them with results from the pilot project (1994-97). MATERIAL AND METHODS: A representative sample of 3,691 students filled in a questionnaire on their smoking habits and attitude to smoking (response rate 75%). Students enrolled in the BE smokeFREE programme were compared with a group of non-enrolled students, and with a third group that reported not knowing whether they were enrolled or not. Information on implementation of the programme was obtained from the first cohort of teachers using the programme who were followed up over three years. The response rate varied from 62% to 65%. RESULTS: From 1997 to 2000, the proportion of Norwegian lower secondary classes enrolled increased from 42% to 62%. Over this period, a total of 110,000 students were enrolled in the programme, receiving on average four periods more of tobacco-related information than students outside the programme. Compared to other tobacco education programmes, BE smokeFREE has been shown to have a significant effect on maintenance of a non smoking lifestyle, number of cigarettes smoked, experiments with chewed tobacco, expectations as to future smoking status and smoke-free behaviour in the school area. For 8th and 9th grade, maintenance of smoke-free status seems to be the same as in the pilot project; for the 10th grade, the results seem to be better. INTERPRETATION: The large-scale implementation of BE smokeFREE has been successful and should be continued. PMID- 11915674 TI - [Smoking is associated with increased risk of ectopic pregnancy--a population based study]. AB - BACKGROUND: There has been a world-wide increase in the incidence of ectopic pregnancies over the last decades. In Norway about 1,500 women are hospitalized every year because of this potentially lethal condition. Several studies have shown that smoking is one of several risk factors for ectopic pregnancies. We wanted to examine whether such an association could be established in a large Norwegian material. MATERIAL AND METHODS: We analysed data from a cross-sectional questionnaire-based study of 9,237 Norwegian women aged 35-49 years. 61.6% responded. RESULTS: 67.4% of the women were present or previous smokers. 301 (3.3%) had experienced an ectopic pregnancy. We found that smokers had a relative risk of 1.5 (95% CI 1.1-1.9) for ectopic pregnancy compared to non-smokers. INTERPRETATION: Our results support other studies. Identification of risk factors is important in the prevention of ectopic pregnancies. Smoking is a risk factor for ectopic pregnancy that can be reduced through counselling. PMID- 11915675 TI - [Dialogue and reflections on difficult questions in health care services]. PMID- 11915676 TI - [Selective serotonin reuptake inhibitors--damages not documented?]. PMID- 11915677 TI - [On bioterrorism]. PMID- 11915678 TI - [The air ambulance service in Bodo]. PMID- 11915679 TI - The rise in bomb threats: how should hospital security respond? PMID- 11915681 TI - Closing the training gap in hospital security. AB - While many healthcare facilities have highly trained security officers, some still relegate security to staff in other full-time positions--maintence, engineering, admissions--and don't provide those individuals with adequate training to perform the security functions they've been assigned. So claims says Linda Glasson, hospital security consultant. Glasson, past president of the International Association for Healthcare Security and Safety (IAHSS), noticed a gap in security training doing consulting work for healthcare facilities. In some instances, inadequately trained staff expressed concern about their ability to perform security functions. Her findings are supported by other hospital security leaders who spell out in this report what adequate training of security and non security personnel should consist of. PMID- 11915682 TI - Making employees the first line of defense in reducing hospital theft. PMID- 11915680 TI - One hospital's experience: improving ER security also extends to patient floors, community. PMID- 11915683 TI - [Myelinated nerve fibers associated with juxtapapillary haemorrhages]. AB - The myelinated nerve fibers (MNF) are o congenital anomaly characterized by the extension of the myelinisation process anterior from the lamina cribosa. The incidence of myelinated nerve fibers is 0.3-0.6% in patients and 0.54% in eyes from autopsies. Although these represent benign lesion, rarely are associated with retinal abnormalities: vascular abnormality (retinal telangiectasis, cranial facial lesions, coloboma of the iris, keratoconus, myopia/strabismus, amblyopia). Myelinated nerve fibers asSociated with juxtapapillary haemorrhages was not reported in the available literature. We describe the clinical findings and diagnostic particularities of myelinated retinal nerve fibers associated with juxtapapillary haemorrhages in a personal case. An 40-year-old woman was referred for unilateral papillary abnormalities. The diagnosis was myelinated retinal nerve fibers as white striated patches with feathery edges, associated with two juxtapapillary haemorrhages. The clinical and pathogenic features are discussed. Based on the association between the myelinated nerve fibers, the juxtapapillary haemorrhages there is a possible pathogenic correlation between these elements. The action of the myelinated nerve fibers can be explained by a mechanical compression, with the disruption of the retinal artery, a structural vulnerable artery. PMID- 11915684 TI - [High-altitude retinopathy]. AB - High-altitude retinopathy is a very rare ocular disease in our country, which can occur isolately or as a part of high-altitude illness. This paper presents the case of a patient with high-altitude illness and the diagnosis and treatment problems of this case. PMID- 11915685 TI - [Rare association--luetic interstitial keratitis and sarcoidosis granulomatous uveitis. Case report]. AB - The goal of this article is to present a clinical case, being particular by the existence of keratitis at a patient who was diagnosed after histological exam of the ocular contents with sarcoidosis granulomatous uveitis. We try to review some aspects characteristic to sarcoidosis, disease which returns in our attention due to the high frequency of uveitis and that's why it deserves a special place in etiology of uveitis. PMID- 11915686 TI - [Ocular signs in Rubinstein-Taybi's syndrome]. AB - Rubinstein-Taybi syndrome, known also as "Broad Thumb-Hallux syndrome", was first recognized by Rubinstein and Taybi in 1963 and is characterized by somatic and ophthalmologic signs. Most commonly somatic changes are: broad thumbs and hallux, craniofacial dysmorphism, growth and psychomotor retardation. Most frequently reported eye anomalies are: antimongoloid slant of the palpebral fissures, strabismus, congenital obstruction of the lacrimal excretory system, colobomas of the iris and of the optic nerve head, ametropia. PMID- 11915687 TI - [The Faden operation in strabismus surgery]. AB - PURPOSE: Is to evaluate the indications of Faden operation and the postoperative results. MATERIAL AND METHOD: We studied a number of 32 patients with convergent strabismus operated in the Universitary Eye Hospital from Cluj, during 1998-2000 at whom we performed Faden operation with conventional surgery on the medial rectus. The anAestesia was with ketamine i.v. RESULTS: The majority of patients where females (62.50%) with ages between 0-14 years. We performed surgery in cases of convergent strabismus with spasm of fixation. The postoperative results were promising; in 23 cases we obtained ortHophory, in 8 cases a remaining deviation angle (5-10 D P), and in 1 case a deviation angle above 5-10 D P. CONCLUSIONS: 1. Faden operations is indicated to correct the spatical component of the deviation angle. 2. The postoperative good results are depending on the aSsociations of Faden operation with conventional surgery. 3. Under or above correction of the strabismus is linked with a deficiency of the conventional surgery associated. PMID- 11915688 TI - [Papilloedema and POEMS syndrome]. AB - POEMS syndrome stands for the first letter of the words: polyneuropathy (P), organomegaly (O), endocrinopathy (E), monoclonal component (M), changes of the tegument (S-skin). In 33-84% of the cases, chronic bilateral papilloedema is associated with the syndrome. The disease is a multisystem disorder, a plasma cell dyscrasia being present, often associated with Castleman disease. Inflammatory cytokines, endothelial vascular growth factor and human herpes virus 8 are thought to play an important role in pathogenesis. This summary is about a 16 year old female patient, where the diagnosis of POEMS syndrome was elaborated based on the systematization of the clinical symptomatology, developed over a long period of time. Based on the latest data from literature, peculiarities of diagnosis, pathogenesis and therapy of the disease are being discussed. PMID- 11915689 TI - [Contraeditorial]. PMID- 11915690 TI - [Bilateral retinal venous thrombosis after 8 years in a young patient with congenital coagulopathy]. AB - The issue discusses the case of a young male patient (35 years old), with bilateral retinal vein thrombosis. The onset of the ocular disease was eight years ago, in the left eye, with decreased visual acuity and complicated with neovascular glaucoma; with an adequate treatment, anatomical and functional evolution was good. A new retinal vein thrombosis, this time in the right eye, make us to concentrate our efforts on the etiology of the disease. Thus, we found a blood hyperviscosity syndrome, caused by a congenital deficiency of protein "S" and resistance at the activated protein "C", which may produce recurrent venous thrombosis. PMID- 11915691 TI - [Current treatment of endophthalmitis]. AB - This paper presents a few theoretical considerations about the causes and treatment of endophthalmitis, about the high role of endophthalmitis vitrectomy. We discuss the evolution and treatment in 3 different clinical cases. PMID- 11915692 TI - [Optical coherence tomography in surgical management of macular diseases]. AB - Optical coherence tomography is a new imagistic technique recently introduced in ophthalmology. It is useful for diagnosis of macular diseases and it is extremely important in the management of those surgical ones. The most suitable diseases for postoperative evaluation by optical coherence tomography are: macular holes, epi and subretinal membranes, macular edema of diverse etiology and severe proliferative diabetic retinopathy. PMID- 11915693 TI - [Electrical changes in the middle retinal layer in diabetic retinopathy]. AB - An electroretinographic study was undertaken on 27 diabetic patients in order to catch the electric changes arose in the mid retinal layer under ischaemic conditions. Significant diminution of the amplitude of the b wave and less significant variation of its implicit time were observed during the progression of the diabetic retinopathy. An unexplained phenomenon--the amplification of the b wave--took place in certain cases of background retinopathy and was emphasised after adaptation of retina to darkness. PMID- 11915694 TI - [Orthoptic treatment efficiency in convergence insufficiency treatment]. AB - We studied a group of 162 patients(89 females, 73 males), with ages between 15-30 years, who complained of blurred vision at near work. 98 patients(60.4%) were diagnosed with convergence insufficiency (C.I.), the rest of 64 patients(39.6%) had: low refractive errors, heterophoria and intermittent heterotropia. Patients with convergence insufficiency were divided in 3 groups: group 1(34 patients- 34.6%) were treated with orthoptic exercises and near point exercises at home, group 2 (34 patients--34.6%) were treated with only near point exercises at home and control group 3 (30 patients--30.8%) without treatment. The result of the treatment of C.I. was good at 25 patients(73.5%) of group 1, at 8 patients(23.5%) of group 2 while in group 3 at only one patient the symptoms disappeared. PMID- 11915695 TI - [High intraocular pressure in patients with hypothyroidism--evolution correlating with serum TSH levels]. PMID- 11915696 TI - [Evolution of ocular infection in diabetes mellitus patients]. AB - PAPER AIM: The paper presents the infectious complications at patients with diabetes mellitus, and the evolution of these complications with the type of diabetes and the treatment with orale hypoglycemic agents or insulin. MATERIAL AND METHOD: The survey was fulfilled in the Clinic of Ophthalmology of Craiova and in the Metabolic Diseases clinic, during a period of 10 years. RESULTS: We surveyed 497 hospitalized patients, and we noticed different infectious complications: kerato-endothelitis in 9 cases, herpetic keratitis in 11 cases, anterior uveitis in 16 cases, corneal ulcus in 24 cases, total uveitis in 6 cases. In 17% of the cases, the patients were not previously aware about the diabetes affection. CONCLUSIONS: The very severe infectious complications of the eye in diabetes, determine the doubling of the ocular and general treatment, for normalisation of the blood glucose and a good evolution of the diabetes. PMID- 11915697 TI - [Pain in Ophthalmology]. AB - The pain, one of the most common and unpleasant symptoms of human experience, is a psychophysiological phenomenon made of physical (elementary) pain and psychological pain. The ocular pain may result from stimulation of trigeminal nerve fibres anywhere within the eye, the surrounding tissues, the deep orbit, and the base of the middle and anterior cranial fossae. An ocular pain which is accompanied by another ocular signs can easily suggest a diagnosis for ophthalmologist, but a deep pain which involves the frontal bone and the deep orbit often asks an interdisciplinary consultation. PMID- 11915698 TI - [Choroidal neovascularization]. AB - Choroidal neovascularization (CNV) may occur in any ocular condition affecting the integrity of the choriocapillaris-Bruch's membrane-retinal pigment epithelium outer retinal layer complex. CNV in the macula is a major cause of severe central visual loss. A variety of clinical symptoms suggest the diagnosis of CNV. Fluorescein and indocyanine green angiography play a very important role in the detection and precise localization of CNV in relation to the centre of the foveal avascular zone. In the present, CNV benefits by many treatment possibilities. The goal of treatment is specifically to decrease the risk of visual acuity loss. PMID- 11915700 TI - [Diagnostic differentiation between two syndromes of progressive myoclonus epilepsy]. AB - The study aimed to differentiate, according to clinical and electroencephalographical criteria, between the most frequent types of myoclonus epilepsy--Unverricht-Lundborg disease (ULD) and Lafora disease (LD). Two patients with ULD and two with LD, aged 13-16 years, have been examined. In all cases, the diagnosis of myoclonus epilepsy has been verified by using molecular genetic analysis. The differential diagnostic criteria have been ascertained for ULD and LD: the earlier age-of-onset in ULD comparing to LD (8.8 and 12.5 years, respectively); tonic clonic paroxysms in ULD and partial ones with visual aura in LD; a presence of nonepileptical subcortical myoclonus, according to EEG data (visual monitoring); negative myoclonus emerging in manifested stage of LD; appearing of regional epileptiform activity for posterior regions on EEG in LD; more rapid progressiveness of extra pyramidal disturbances with organic dementia outcome in myoclonus LD. PMID- 11915699 TI - [Eye disease in painters--Camille Pissaro]. AB - Some vision disorders to painters have influenced their work. Camille Pissarro the dean of impressionism, presented from forty-eight years dacryocystitis on his left eye and ipsilateral conjunctive chemosis temporally. In that time the treatment consisted in incision and drainage of the lacrymal sac, and cauterization. Due this chronic disease he was forced to paint inside, behind closed windows, in order to avoid dust and wind. This situation influenced the subjects of his painting. His painting reflects large crowds Paris avenues and buildings. This is characteristic for his wonderful works. PMID- 11915701 TI - [Multifactorial nature of Chiari type I malformation]. AB - The aim of the study was to analyze genetic aspects of Chiari type I malformation (CMI) using clinical neurological examination and magnetic resonance imaging data on the craniovertebral region in 13,500 outpatients, 364 inpatients with CMI and 91 first-degree relatives of 47 probands with CMI. Higher CMI frequency (3.9%) in the outpatients with neurological signs, and CMI clinical and morphologic polymorphism have been discovered, the women being mostly affected and the men displaying more severe clinical picture of the disease. In the group of patients with the most severe CMI form, higher parent's endogamy marriage frequency was observed. The CMI clinical and morphologic features in the probands and their relatives were identical and the frequency of the CMI presence in the first degree relatives depended on the proband's pathology severity. The results obtained support the evidence for CMI multifactorial nature. PMID- 11915702 TI - [Obsessive-phobic disorder typology in schizophrenia]. AB - The study aims at clinical characteristics of obsessive-phobic disorders (OPD) in schizophrenia with regard to their modification in main disease development and interaction with basic schizophrenia symptoms (positive and negative). Eighteen patients, mean age 28.2 years, were examined. An affinity degree of schizophrenia to OPD was found to be associated with endogenous progressive process. In slowly progressive schizophrenia, an affinity is selective, concerning to one of the obsessive phobic syndrome component (phobic component proper or avoidance behavior) and realizing in either positive, or negative manifestation of schizophrenia. Two obsessive-phobic syndrome types were determined: OPD of the 1st type--panic and generalized anxiety disorder (52 patients) are modified because of overlapping by chronic endogenous anxiety, obsessive-phobic symptoms being not represented in structure of negative changes (asthenia defect); in OPD of the 2nd type (agora and social phobias, 80 patients) an overlapping of psychopathological symptoms related to avoidance behavior and negative changes (psychopathic-like defect "verschroben" type) was found. Phobic syndrome component does not tend to psychopathological transformation and undergoes reverse changes. In typical cases of progressive schizophrenia has no affinity. But exclusion is--the OPD of the 3rd type (obsessive-compulsive disorders with a fear of external threat and massive defense rituals, 49 patients). In this case, total affinity to schizophrenia is realized in overlapping of both obsessive syndrome components with positive and negative manifestation of schizophrenia. PMID- 11915703 TI - [Contribution of genetics to medicine]. PMID- 11915704 TI - [Psychotic disorders of sexual and erotic content in different diseases]. AB - The results of long-term (1970-2000) clinical-psychopathological study of 251 patients with different forms of schizophrenia (161 cases), manic-depressive illness (19) and exogenous organic mental disorders (71) are summarized. The dynamics and different manifestations of psychotic disorders of sexual and erotic content described. These disorders were appeared mostly by hallucinations and delusions of various content. Similarity and difference of sexual and erotic symptoms and syndromes for different mental diseases have been determined. The psychoses studied were found to manifest more often in persons with different types of abnormal personality and in those with family history positive for mental disorders as well as in the presence of exogenous pathogenic factors. PMID- 11915706 TI - [Diagnostic significance of sleep EEG in children with febrile seizures]. AB - To obtain effective criteria for febrile seizures (FS) differential diagnosis and prognosis, clinical examination and EEG recorded during wakefulness and daytime sleep were studied in 75 FS children and 15 controls aged from 6 months to 6 years. According to duration, frequency and complications of FS, all patients were divided into three groups: "simple" FS (n = 35), "complex" FS (n = 25) and "afebrile seizures" (AFS), i.e. those who developed non-febrile seizures after febrile ones (n = 15). In waking state EEG, epileptic activity was found in only 60% of AFS patients. During sleep stages I-III, in 84% of the patients with complex FS as well as in all AFS patients generalized discharges of (poly) spike waves and (or) focal spikes were detected. Paroxysmal changes, atypical scalp distribution, vertex-potentials and sleep spindles generalization as well as insertion of spikes into arousal-rhythms were determined as additional criteria for unfavorable FS course. In the patients with main features of sympathetic adrenal reaction during febrile paroxysms, complicated FS course was not found. EEG examination during sleep in children with FS increases an efficacy of differential disease diagnosis. Epileptic discharge detection, along with exposure of paroxysmal transformation of physiological sleep EEG transients, that may be earlier sign of epileptic dysfunction, are recommended for proper prevention and treatment of FS. PMID- 11915705 TI - [Intravenous spondylo-infusion in the treatment of lumbar osteochondrosis]. AB - Using the intraosseous infusion technique, 87 patients with neurological symptoms of osteochondritis of lumbosacral intervertebral disks (study group) were treated. Sixty-six patients were treated by traditional drugs (control group). The study and control groups have been matched by age (mean 45 years) and symptoms--reflex-muscular, spondilosis radicular, Charcot's syndrome. An estimation of treatment efficiency was carried out by registration of somatosensory evoked potentials and ultrasonic dopplerography of the inferior vein cava and its branches as well as by measuring of intraosseous pressure in an acantha before and after infusion of a solution. The diskogenic neurological frustrations regressed more promptly and completely in the patients, to whom the intraosseous infusion technique has been applied. The positive correlation between clinical recovery and a decrease of a latency of somatosensory evoked potentials peaks is revealed. The intraosseous pressure in an acantha after a course intraosseous infusion was reduced. The directions for applying intraosseous infusion technique are developed. PMID- 11915707 TI - [The application of parametric statistical methods for non-numerical data in psychiatry]. AB - Impossibility the application of arithmetical (algebraic) calculations and Student's t-criterion for score valued analysis is discussed. The method adequate for score valued data is suggested, being applied, as an example, to Hamilton depression scale and to estimation of scientific publication quality. PMID- 11915708 TI - [Clinical polymorphism, genetic heterogeneity and primary myopathy pathogenesis problems]. AB - The peculiarities of clinic picture of primary myopathies with general gene defects, in particular differences in disease debut, dystrophy severity and degree of diverse muscle group's involvement in the dystrophic process, were analyzed. An attention was drawn specially to the cases of similar and identical mutations, including family cases, which gave rise to dramatically different pathological phenotypes. This information, along with the Becker muscular dystrophy cases (with deletions of DMD gene), describing the patients, aged 55 to 60 years, who maintained a certain movement activity, allow to revise an absolutely severe prognosis of primary myopathies and to stimulate primary myopathy pathogenesis studies. Hypothesis of "second" factors in primary myopathy's pathogenesis is discussed. PMID- 11915709 TI - [A role of social fostering for rehabilitation of mental patients in nursing homes]. AB - A number of younger disabled persons, aged 18-41 years, who are staying in social care units--psychoneurological nursing homes of the Russian Federation Ministry for Labor and Social Development--are reported to be 42%. A rehabilitation effect rise for these patients and their integration with community on the basis of communicative skill and professional training development proves to be important. A Kurov (Moscow region) psychoneurological nursing home activity is suggested as an example for partial resolution of the problem in the form of "home visits" for the most adapted patients and they stay for the period from 1-2 days to 3 months in the rural families--those of relatives, friends, nursing home staff--for supporting, training and work. A development of different daily living skills (buying, public transport using, etc) was higher in those, staying in the families, as compared to the control group. The problem for fostering law renewing and improvement is discussed. PMID- 11915710 TI - [Psychiatric service in general hospital]. AB - The article summarizes the 3-year (1998-2000) consulting and treatment experience of a psychotherapeutic unit in one Moscow general hospital. Psychiatrists realized emergent and planned consultations of the mentally-ill patients in all hospital departments, conducted, in addition to general treatment, psychopharmacotherapy and psychotherapy or directed them, on demand, to psychiatric hospitals. For the 3-year period, the psychiatrists have consulted 4685 somatic patients with comorbid mental diseases (5% of the total amount of the patients admitted to the hospital at that period), 1360 patients (29%) being affected with psychotic disorders. One hundred eighty two patients were diagnosed to have schizophrenia, 784--arteriosclerosis and old-age psychosis and dementia, 45--reactive psychosis, 16--epilepsy (disphoria and psychotic episodes), 67- mental retardation with inadequate behavior, 266--delirium of alcoholic and drug addiction genesis. In some cases, usage of psychotropics, in combination with psychotherapy, has resulted in recovery or considerable psychic state improvement in 9.1% of the patients, improvement in 52% ones, insignificant improvement in 36%. In 2.9% of the cases, the effect of treatment has not been achieved. PMID- 11915711 TI - [The treatment of Parkinson disease]. PMID- 11915712 TI - [Modern views on etiology and pathogenesis of herpes infections of central nervous system]. PMID- 11915713 TI - Professionals and post-hospital care for older people. AB - Hospital elderly care teams in England assess whether patients need assistance, such as community or residential care, upon discharge from hospital. Does the type of multidisciplinary team influence post-hospital care decisions? The aim of this study was to identify which factors predict the services that older people receive upon discharge from hospital. Three multidisciplinary teams were compared where different professionals took the lead in co-ordinating the care assessment process. Data were collected in a case note review of patients (n = 456) aged 75 years and over: patient characteristics, referral patterns, and the types of service received in the month after leaving hospital. Multivariate statistical analysis (controlling for patient characteristics) showed different predictors for different post-hospital services. The nurse-led team was least likely to refer patients for care assessment and these patients received the least post hospital services. The occupational therapy-led team arranged the most occupational therapy services and equipment and their patients received the most home care. The social work-led team referred the most patients for care assessment and their patients received the greatest range but not the greatest amount of services. These results suggest that multidisciplinary team configurations influence post-hospital services for older patients, reflecting professional preoccupations as much as patient care needs. PMID- 11915714 TI - Role overlap between occupational therapy and physiotherapy during in-patient stroke rehabilitation: an exploratory study. AB - The concept of role overlap between occupational therapy and physiotherapy has been the subject of debate for at least three decades. Stroke rehabilitation is an area where role overlap between occupational therapists and physiotherapists occurs. This article reports an exploratory study carried out with nine physiotherapists and nine occupational therapists working in a variety of in patient stroke rehabilitation settings. Analysis of qualitative data collected through semi-structured interviews revealed that the majority of the participants recognised the existence of role overlap as inevitable within collaborative health care and felt it was of benefit to patients. However, it appeared that the concept was also perceived as a challenge to role security by many when considered from a professional perspective. Acceptance of role overlap depended upon the extent to which it occurred in the particular setting. Generic therapy was seen as an extreme form of overlap and regarded as an undesirable progression by most participants. The main strategy used to challenge this development was to emphasise professional uniqueness through role delineation. However, this strategy was found to be weak in the context of increasing demands for collaboration at a policy level. PMID- 11915716 TI - Ten years on. PMID- 11915715 TI - The London training ward: an innovative interprofessional learning initiative. AB - This paper reports the findings from an evaluation of a pilot interprofessional training ward project for pre-qualification medical, nursing, occupational therapy and physiotherapy students. This initiative required sustained collaboration from staff based in two National Health Service (NHS) trusts and four schools in three universities. The ward was based on a model of interprofessional education developed in Sweden, but adapted in the light of this experience and also to meet the needs and aspirations of the training ward stakeholders in London. The training ward was evaluated using a multi-method design. Data were collected from all participants involved in this pilot: students, facilitators and patients. The findings from the evaluation are presented and discussed. This paper pays particular attention to the collaborative experiences of the students, staff and institutions involved in this initiative. In addition, 1-year follow-up data collected from the students who had, by then, qualified as clinical practitioners are reported. The paper presents conclusions from the evaluation, and comments on the training ward's strengths, limitations and future development. PMID- 11915717 TI - Using collage art work as a common medium for communication in interprofessional workshops. AB - This paper explores the notion that interprofessional working is often hampered by the lack of a common language set and the variety of language sets used across the caring professions. The use of art as a common ground communication tool is presented with particular reference to the medium of magazine picture collage work. The paper describes how the use of this medium can enhance the understanding of individuals in interprofessional study days and workshops, where particular issues can be presented and explored by the participants, resulting in the increased awareness of the different perceptions of shared issues. In this way the use of magazine picture collage is advocated as a communication tool aimed to build bridges across the range of language sets used by different professionals. PMID- 11915718 TI - Perceptions regarding the use of physical restraints with elderly persons: comparison of Israeli health care nurses and social workers. AB - In view of the difficulty involved in decision-making regarding the use or removal of physical restraints and the recent pattern encouraging the use of interdisciplinary teams for elder care issues, the present study compared the perceptions of Israeli nurses and social workers in health care settings regarding the use of physical restraints. Data were collected from a convenience sample of 50 nurses and 69 social workers working in long-term and acute care settings. The findings indicated that participants in all professions attributed moderate to low importance towards the use of physical restraints. Social workers' perceptions were similar to those of nurses in psychiatric hospitals and slightly more favourable to the use of physical restraints than those of nurses in nursing homes. Patients' safety (as reflected in the scores of the items related to protecting an older person from falling and protecting an older person from pulling out a catheter) was the most important reason for using physical restraints for both groups. Increased attention should be given to the role of social workers as team members in the process of decision-making regarding the use or removal of physical restraints, especially as mediators between the elderly person, family members and staff members. PMID- 11915719 TI - A portrait of clinical psychology. AB - The purpose of this paper is to provide an introduction to the profession of clinical psychology. We provide a definition and overview of the core features of the profession, followed by a description of entry requirements and education. We go on to describe the dominant models which have driven the training and practice of clinical psychologists through the latter half of the 20th century to the present day. We conclude with a look to the future of the profession and a brief consideration of opportunities and threats. Throughout we have attempted to pick out aspects of the profession that are relevant or of interest, irrespective of national differences in professional training and practice, or variations in the development of clinical psychology in particular nation states. When generalisations would be too broad to be useful or are otherwise difficult, we have provided specific information about the profession as it has developed in the UK or USA. The references should provide a rich source of further information for the interested reader. PMID- 11915720 TI - Interprofessionality in health and social care: the Achilles' heel of partnership? AB - Partnership working is now a central plank of public policy in the UK, especially in the field of health and social care. However, much of the policy thrust has been at the level of interorganisational working rather than at the level of interprofessional partnerships. The empirical and theoretical literature is largely sceptical about the feasibility of effective joint working between separate but related professionals--the 'pessimistic tradition'. Based upon an empirical study of general practitioners, community nurses and social workers in northern England, this article challenges such a tradition and proposes an 'optimistic hypothesis' for further investigation. PMID- 11915721 TI - Delivering health and social care: changing roles, responsibilities and relationships. PMID- 11915722 TI - Systematic reviews of evaluations of interprofessional education: results and work in progress. PMID- 11915723 TI - Meeting highlights--International Consensus Panel on the treatment of primary breast cancer. AB - The 7th International Conference on Adjuvant Therapy of Primary Breast Cancer, with several thousand delegates from 70 countries, was held in February 2001 in St. Gallen, Switzerland. Its consensus recommendations were summarized in the Sept. 15 issue of the Journal of Clinical Oncology (Vol. 19, No. 18, 2001: pp 3817-3827). The panel conference developed guidelines advising that all patients with endocrine-responsive tumors receive anti-hormonal therapy, including those at minimal risk of recurrence (defined as a 10% risk at 10 years). The panel further lowered the threshold defining endocrine-responsive disease to tumors containing as few as 1% of cells that stain positive for steroid hormone receptors. Treatment of primary breast cancer should depend on the hormonal status of tumors. The last set of guidelines, issued in 1998, supported a more limited use of endocrine-based therapy, but recent research has convinced the oncology community otherwise. Postmenopausal women have the greatest prevalence of hormone-positive tumors, and more than one-third of breast cancers in women of child-bearing age are estrogen dependent. The panel also highlighted several other important advances, including: the importance of factoring patient preferences into treatment decisions; the value of sentinel lymph node biopsy in avoiding extensive surgery; and incorporation of new agents and improved outcome in the treatment of breast cancer. PMID- 11915724 TI - [Present and future in the surgical treatment of pancreatic cancer]. AB - Owing to the recent advances in surgical techniques and postoperative care, the operative indication for adenocarcinoma of the pancreas has been widened in Japan. In our institution, we have extended lymphatic and connective tissue clearance during surgical resection of this cancer. As a result, the 5-year survival rate has increased from 8% to 25% due to a significant decrease in the incidence of locoregional recurrence. The prognostic benefit gained by this procedure is seen mainly in those patients with stage III cancer or in whom no positive nodes were detected beyond the pancreatic head region. Thus, our extended surgery seems to have been useful due to eradicating the microinvasion in the peripancreatic connective tissues, rather than dissecting the positive lymph nodes. More advanced cancers should be treated by effectively combining chemo- and/or radiation-therapies with surgery. In order to cure the patients more easily with less-invasive surgery in the future, we should develop both an early diagnosis system and sensitive examinations of micrometastases or microinvasions. PMID- 11915725 TI - [Current status of adjuvant therapy for pancreatic cancer]. AB - Pancreatic cancer is considered to be one of the malignancies most resistant to therapy. It is characterized by early local invasion and distant spread. Therefore, resection with curative intent is limited to a very small proportion of patients. Even in these selected patients, long-term survival remains very poor because of liver and local recurrence. Therefore, control of occult liver metastasis and local residual tumor with perioperative radiotherapy and chemotherapy may provide some palliative benefits, and should have some impact on overall survival. However, none of the studies to date are considered definitive. Japanese pancreatic surgeons have developed a number of adjuvant therapies which theoretically could be good enough to prolong long term survival, however, they have not been tested in randomized controlled trials. Planning co-operative studies on this important issue in pancreatic cancer therapy is urgently needed. PMID- 11915726 TI - [Radiotherapy and chemotherapy for unresectable pancreatic cancer]. AB - Despite many advances in the diagnosis of pancreatic cancer (PC), only a small minority of patients are candidates for surgical resection with curative intent. Moreover, even for these selected patients, the prognosis remains unfavorable because of postoperative recurrence. Accordingly, to improve the prognosis of PC, there is an urgent need to develop effective non-surgical treatments for this disease. Radiotherapy (RT) alone does not have a significant clinical impact in PC, but when combined with chemotherapy (CHT), it provides a survival benefit in locally unresectable PC. However, to establish optimal methods of combining RT with CHT, newer approaches such as specialized RT techniques and new CHT agents are actively being pursued. At present CHT for PC has only limited value in clinical practice due to the lack of any individual agent with a truly high level of activity. The strategies for effective CHT in PC include biochemical modulation of anticancer agents, multi-agent CHT, and new agents with different mechanisms of action. Among these strategies, the identification of more effective new agents is a high priority in developing a truly effective CHT for PC. The novel nucleoside analog gemcitabine has the potential to produce better results than those achieved with 5-FU, and has been accepted as first-line CHT for patients with advanced PC. Moreover, increased understanding of the biology of PC should facilitate the development of entirely novel treatment options. We must continue to actively search for more effective non-surgical treatments for PC. PMID- 11915727 TI - [A new regional arterial infusion chemotherapy for patients with advanced pancreatic cancer]. AB - Various arterial infusion chemotherapies have been tried for the purpose of local control of advanced pancreatic carcinoma. However, these treatments were not effective against the primary lesion because of its special anatomical position and the complex hemodynamics, although they were effective against the liver metastases. Therefore, the vascular supply distribution was altered by superselective embolization to control the primary legion in the pancreas, after transcatheter peripancreatic arterial embolization toward the primary site. Furthermore, bilateral (hepatic and splenic) arterial infusion chemotherapy was applied to both the primary site and liver metastasis. As a result, the response rate was 73.9%, with a mean survival period 18.26 +/- 10.06 months. We believe that the current chemotherapy was an effective treatment for unresectable pancreatic cancer since it was possible to treat patients with little harm to their quality of life. PMID- 11915728 TI - [Immunotherapy for unresectable pancreatic cancer]. AB - To treat unresectable pancreatic cancer, we have performed cell therapies using lymphokine-activated killer cell, cytotoxic T lymphocytes (CTLs) induced by mixed lymphocyte culture (MLC) with autologous tumor cells, and CTL recognizing MUC1 induced by MLC with an allogeneic pancreatic tumor cells line, YPK-1. CTL therapy was effective in some cases. We also performed cell therapy using MUC1 peptide pulsed dendritic cells (MUC1-DCs) and MUC1-CTLs. This therapy was effective in one of three cases so far. Cancer peptide vaccine therapy trials for unresectable pancreatic cancer are also ongoing. Cell therapy and peptide vaccine therapy may be promising approaches for unresectable pancreatic cancer. PMID- 11915729 TI - [Gene therapy for pancreatic cancer]. AB - In order to develop a new therapeutic intervention for pancreatic cancer, we have examined the effect of gene therapy for pancreatic disease. The transfection of the gene for UPRT, a 5-FU-converting enzyme, resulted in a significant change in the sensitivity of pancreatic cancer cells against 5-FU, resulting in the decrease of the tumor volume disseminated in the abdominal cavity of mice. Although the production levels of vascular endothelial growth factor (VEGF) in pancreatic cancer cell lines are different, anti-angiogenesis gene therapy using a soluble form of VEGF receptor (flt-1) has been demonstrated to be a promising strategy for pancreatic cancer. The transfection efficacy is the crucial point for the success of gene therapy; therefore, it is necessary to develop a vector system for solid tumors. It has been revealed that replication-competent adenoviruses are not only a strong weapon themselves, but are also useful carriers of genes possessing anti-tumor activities as virus vectors specific to tumors without normal p53 function or intact Rb pathway. Determining whether these experimental results are universally true will require clinical trials in the future. PMID- 11915730 TI - [Phase II study of combined Vinorelbine (Navelbine) plus cisplatin for advanced non-small cell lung cancer (an interim report)]. AB - A phase II study of a combination therapy with vinorelbine (Navelbine) plus cisplatin was performed using 27 patients with advanced non-small cell lung cancer, to evaluate the efficacy and safety of this therapy. Patients were treated, at 3-week intervals, with two or more courses of a regimen consisting of vinorelbine 25 mg/m2 (days 1, 8) and cisplatin 80 mg/m2 (day 1). The response rate was 48.1% (13/27) (95% confidence interval: 28.7-68.1%). The median number of treatment courses was 2.6. The major toxicity was hematological: 74% of patients had neutropenia of grade 3 or higher that subsided rapidly upon administration of a granulocyte-colony stimulating factor. Non-hematological toxicities were mild, of grade 2 or less: the main symptom was nausea/vomiting (62%), although constipation, eruption, anorexia, peripheral nerve disorders, diarrhea, fever, and other conditions were also observed. In conclusion, the high response rate and good tolerance to this combination therapy warrants further study. PMID- 11915731 TI - [Correlation between clinical pathophysiologic factors and expression of orotate phosphoribosyl transferase (OPRT), thymidylate synthase (TS), and dihydropyrimidine dehydrogenase (DPD) in colorectal cancer]. AB - We measured the activity of orotate phosphoribosyl transferase (OPRT), the amount of thymidylate synthase (TS) enzyme, and the activity of dihydropyrimidine dehydrogenase (DPD) for individual tissue types in order to study the contribution of these substances to the effects of the pyrimidine fluoride anticancer drug 5-fluorouracil (5-FU). We also studied the correlation between these 3 enzymes and clinical pathophysiologic characteristics (age, sex, extent of tumor invasion, extent of metastasis to the lymph nodes, lymphatic invasion and the venous invasion of the colorectal wall). Sixty-eight patients with colorectal carcinoma who had undergone surgical resection in our department were studied. There was a significant (p < 0.01) elevation of OPRT activity in the tumor tissue compared with regions of normal tissue. OPRT activity levels in the tumor tissue were lowest in patients with mucinous carcinoma while TS enzyme levels showed the highest activity in tumor tissue in poorly differentiated adenocarcinoma. DPD also showed high activity levels in tumor tissue in poorly differentiated adenocarcinoma and mucinous carcinoma. It is possible that the expression of enzymes with respect to the antitumor effects of 5-FU is a factor contributing to the poor prognosis for patients with poorly differentiated adenocarcinoma and mucinous carcinoma. In the present study of clinical pathophysiologic characteristics, we found that metastasis to the lymph nodes was associated with a significant reduction in the OPRT tumor/normal (T/N) ratio. Our results indicate that it may be possible to predict lymphatic metastasis by determining the T/N ratio for OPRT before surgery. PMID- 11915732 TI - [Combination chemotherapy with nedaplatin (CDGP) and 5-FU for oral cancer]. AB - Chemotherapy using CDGP plus 5-FU was evaluated in patients with oral cancer. The subjects were patients with squamous cell carcinoma of the oral cavity who had not received any therapy, comprising 7 patients with carcinoma of the tongue, 2 with buccal carcinoma, 2 with maxillary gingival carcinoma, and 1 with carcinoma of the oral floor. There were 4 patients in Stage II, 3 patients in Stage III and 5 patients in Stage IV. Patients with a PS < or = 1, WBC > or = 4,000/mm3, Hb > or = 10 g/dl, platelet count > or = 10 x 10/mm3, and normal liver, kidney, and heart function at baseline were selected for this study. In all patients, 5-FU was administered at a dose of 600 mg/m2/day for 5 days (day 1 to day 5) by continuous infusion, for a total dose of 3,000 mg/m2. CDGP was administered on day 1 at a dose of 80 mg/m2 in 8 patients and at 100 mg/m2 in 4 patients. This treatment was one course of therapy, and patients received 1 or 2 courses. Of 12 patients who were evaluable, there were 9 partial responses and 3 no changes, for a major response rate of 75%. Toxicities experienced by patients were mild (grade 2 or lower) gastrointestinal disorders (including nausea/vomiting) and renal impairment, while grade 3 leukopenia and thrombocytopenia developed in 1 patient each and grade 4 thrombocytopenia occurred in another patient. Thus, patients receiving CDGP + 5-FU therapy should be closely monitored for hematologic toxicity. Since CDGP + 5-FU therapy achieved a good response rate (75%) in the treatment of squamous cell carcinoma of the oral cavity, we plan to use this therapy in the future and assess its benefit in a larger number of patients. PMID- 11915733 TI - [Successful treatment of advanced recurent breast cancer using DMpC therapy as maintenance therapy]. AB - A 65-year-old female who underwent radical mastectomy on 28 April 1988 felt swelling and numbness of her left lower jaw. As a result of bone biopsy and scintigraphy, multiple bone metastasis was diagnosed. After 6 cycles of CMF therapy, the swelling and numbness of the left lower jaw were reduced, but atelectasis of the left lung upper lobe appeared. After 10 cycles of paclitaxel therapy, atelectasis and bone metastasis were reduced. Daily oral chemoendocrine combination therapy, DMpC therapy (5'-DFUR 800 mg/day + CPA 100 mg/day + MPA 800 mg/day) was continued for 12 months. No recurrent signs and serious side effects were observed during DMpC therapy. PMID- 11915734 TI - [A case of recurrent lung cancer successfully treated using ambulatory chemotherapy with vinorelbine]. AB - The patient was a 64-year-old woman. She had previously suffered from stage IA lung adenocarcinoma and undergone a right upper lobe resection. After 7 years, she complained of coughing and right chest pain. Right pleural effusion and tumor mass shadow/lymph node swelling on the surgical margin of a right bronchus were found. A tumor marker (CEA) was also at a high level. Combination chemotherapy with cisplatin (CDDP) and docetaxel (TXT) failed to produce an antitumor effect. The treatment was changed to ambulatory chemotherapy with vinorelbine (VNB) at 20 mg/m2 weekly. This produced tumor reduction and disappearance of the pleural effusion, and normalized the CEA level. No adverse reactions except leukopenia of grade 2 occurred. VNB alone seemed to successfully exhibit an antitumor effect. As VNB is easy to administer, it will be applicable in ambulatory chemotherapy administered with consideration of the patients quality of life. PMID- 11915735 TI - [Chemotherapy for small-cell lung cancer (SCLC) patients with renal failure]. AB - We administered chemotherapy in three cases of small-cell lung cancer (SCLC) with renal failure under different situations. Hemodialysis (HD) was used in 2 out of the 3 cases. Case 1 was complicated by acute renal failure from extensive bilateral tumor invasion. After chemotherapy (CBDCA + ETP) under HD, renal metastases regressed and renal function improved, although the final response was PD. In case 2, HD had been introduced for diabetic nephropathy. After 2 cycles of chemotherapy (CBDCA + ETP) under HD, the patient attained a PR. Case 3 is an example of paraneoplastic nephrotic syndrome with renal failure. Chemotherapy including CBDCA or CDDP was performed and the QOL of the patient improved. Pro GRP and serum creatinine changed in parallel during the clinical course of 6 admissions. In conclusion, individualized therapy is necessary to increase survival time of SCLC patients with renal failure. Although chemotherapy is useful, further study is needed for the selection of suitable chemotherapeutic regimens, optimal dosage of each drug and the timing of HD. PMID- 11915736 TI - [An elderly patient with advanced non-small-cell lung cancer that responded remarkably to combination chemotherapy of vinorelbine and gemcitabine]. AB - A 73-year-old man was admitted to our hospital because of hemoptysis in November 1999. A chest CT revealed a mass shadow in the right upper lobe and enlarged hilar and pretracheal lymph nodes. Bone scintigraphy showed a bone metastasis in right middle rib. He was treated with vinorelbine (25 mg/m2 days 1,8) and gemcitabine (800 mg/m2 days 1,8) every 3 weeks from December 1999. After 4 courses of chemotherapy, the primary tumor and hilar lymph node were remarkably reduced in size, and his hemoptysis disappeared and body weight increased. It is suggested that combination chemotherapy with vinorelbine and gemcitabine is effective for elderly patients with non-small-cell lung cancer with good performance status. PMID- 11915737 TI - [A patient with advanced gastric cancer in the gastric tube whose QOL was improved by TS-1]. AB - We present the case of a 72-year-old man with gastric tube cancer accompanied by multiple liver metastases, after esophagectomy for esophageal cancer, whose quality of life (QOL) was improved with a small dosage of TS-1. The patient's high serum AFP level suggested alpha-fetoprotein-producing gastric cancer. He was treated with half the standard dose of TS-1, because the patient's poor general condition necessitated chemotherapy with low toxicity and high efficacy. The daily dose was 40 mg for the first three courses and 50 mg for the last two. Each treatment course consisted of a four-week administration followed by two drug free weeks. The patient received five courses of chemotherapy at our outpatient clinic before his death from re-progression of liver metastasis. No serious side effect except temporary stomatitis was observed. A decrease in tumor markers, alpha-fetoprotein and carcinoembryonic antigen, was obtained after 4 weeks. After 2 cycles, computed tomography and endoscopy examinations showed regression of the primary tumor and liver metastases, and tumor markers were decreased remarkably. The patient's QOL improved gradually after the treatment. His performance status before the chemotherapy was 3, and improved to 1 after two cycles. The small dosage of TS-1 was effective without any adverse effects, and improved the patient's QOL, for 6 months. PMID- 11915738 TI - [A gastric cancer patient with marked para-aortic lymph node metastasis surviving more than 10 years after aggressive operation and repeated EAP chemotherapy]. AB - The high incidence of side effects for EAP (etoposide, adriamycin, cisplatin) combination chemotherapy led to the recent decline in its use. However, we report herein the long-term disease-free survival of a woman following postoperative EAP therapy. A 57-year-old woman was referred to our hospital because of general malaise. X-ray and endoscopic examination revealed a Borrmann type 3 gastric cancer. Preoperative computed tomography and ultrasonography revealed multiple para-aortic lymph node swellings. The patient simultaneously underwent subtotal gastrectomy and splenectomy, and complete para-aortic lymph node dissection. Histopathological tests revealed that the tumor was a poorly differentiated adenocarcinoma with 35 metastatic para-aortic lymph nodes. The patient was treated with 2 cycles of EAP therapy. After discharge, swelling in one para aortic lymph node was detected. Following three subsequent cycles of EAP therapy, the swollen lymph node disappeared and the patient has remained disease free for 10 years. This case illustrates that aggressive surgery followed by repeated courses of EAP therapy can produce excellent clinical outcomes. PMID- 11915739 TI - [Complete response in a case of simultaneous esophageal and gastric cancer treated by combined radiotherapy and chemotherapy of TS-1 and CDDP]. AB - A 75-year-old male patient was hospitalized for examination of abdominal trouble. Endoscopic examination simultaneously revealed 0-I pl + II c esophageal cancer and type 3 gastric cancer. Endoscopic treatment is impossible to resect the lesion completely. It is difficult for the patient to undergo an operation because he has suffered from a cardiac infarction and pulmonary trouble. Thus, gastric cancer was treated by chemotherapy and esophageal cancer by concurrent chemoradiotherapy with chemotherapy used for gastric cancer. Chemotherapy consisted of CDDP and TS-1 every 4 weeks. TS-1 (100 mg/day) was administered on days 1 to 21, and CDDP (70 mg/m2) was infused for 24 hours on day 8. Radiotherapy (5 days/week) at 2 Gy/day was concurrently started from the beginning of chemotherapy. At day 8 in the 2nd course of chemotherapy, leucocytopenia of grade 2 and appetite loss of grade 3 (NCI-CTC) were seen. Chemoradiotherapy was then suspended for one week. After recovery from toxicity, treatment was continued for a week. A total of 64 Gy was administered. After treatment, endoscopic examination with biopsy revealed the disappearance of gastric and esophageal cancer. PMID- 11915740 TI - [A case of hepatic metastasis from rectal carcinoma successfully treated with pharmacokinetic modulating chemotherapy (PMC)-CPT-11 therapy]. AB - We performed PMC-CPT-11 therapy (modified pharmacokinetic modulating chemotherapy plus irinotecan, or modified PMC) in a case of sigmoid colon cancer with local invasion and multiple hepatic metastases. This regimen combined PMC therapy which includes Hartmann's operation, simple hysterectomy and postoperative 5 fluorouracil (5-FU), with intravenous irinotecan, or CPT-11. The multiple hepatic metastatic lesions disappeared after surgery and no local recurrence has been found since. These results indicate that PMC-CPT-11 (modified PMC) therapy could be an effective regimen for cases of progressive colon cancer in the future. PMID- 11915741 TI - [Combined preoperative therapy for oral cancer with nedaplatin and radiation]. AB - We performed preoperative combined therapy using nedaplatin (CDGP) and radiation in 12 patients with squamous cell carcinoma originating from the oral cavity and maxillary sinus, and examined for any adverse events that may have occurred during this therapeutic regimen. Regarding the irradiation, external irradiation utilizing a 6 MV linac (linear accelerator) at a dose of 2.0 Gy/day was performed 5 times a week, with the target total radiation dose set at 40 Gy. In addition, CDGP was intravenously administered 30 minutes before irradiation at a dose of 5 mg/m2/day. Mucositis was observed in all 12 subjects, however, the severity was observed to be grade 1-2 with no major differences in comparison to the patients given standard radiation monotherapy. Two subjects developed grade 3 leucopenia and were thus given granulocyte colony stimulating factor (G-CSF). In addition, grade 2 and grade 3 thrombocytopenia were both observed in one subject each. The subject with grade 3 thrombocytopenia required a platelet transfusion during surgery. No marked changes in serum creatinine levels were noted. These findings are therefore considered to provide evidence supporting the safety of this combination therapy. PMID- 11915742 TI - [A case of subacute transient cerebral dysfunction in a osteosarcoma patient following high-dose methotrexate]. AB - Subacute transient cerebral dysfunction following high-dose methotrexate occurred during neo-adjuvant chemotherapy for an eighteen-year-old male with osteosarcoma in his right femur. The variety of symptoms including hemiparesis and hesitancy of speech occurred 8 days after an administration of high-dose methotrexate (10 g/m2). Evaluations including CT scan of the brain, hemogram and blood chemistry revealed no abnormal findings. The patient found it difficult to sit, speak and eat, but was not confused. He improved in a few days without any specific treatment or residual sequelae. This syndrome was transient and did not recur when he had two additional HD-MTX treatments. The cause of this syndrome remains unknown and does not seem to be predictable. It is, thus, necessary for oncologists to take due care with regard to this syndrome in HD-MTX chemotherapy. PMID- 11915743 TI - [Rituximab, a chimeric anti-CD20 monoclonal antibody, in the treatment of B-cell lymphoma]. AB - In September 2001, rituximab, a chimeric mouse-human anti-CD20 monoclonal antibody, was approved for the treatment of B-cell lymphoma by the Japanese government. Rituximab is the first monoclonal antibody that was approved for the treatment of malignant neoplasms by the U.S. Food and Drug Administration (FDA). Several clinical trials of rituximab conducted in USA, Europe and Japan revealed its promising therapeutic efficacy for B-cell lymphoma. Its minimal myelotoxicity allows rituximab to be combined with full doses of anticancer agents. Ongoing clinical trials will define the future role of rituximab in the treatment of B cell lymphoma. PMID- 11915744 TI - [Tumor marker in ovarian cancer]. AB - The current role of tumor markers in the management of ovarian cancer is reviewed. The most useful tumor marker in epithelial ovarian cancer is still the antigen CA125. However, the level of this marker is modified by peritoneal irritation from endometriosis or inflammatory disease. Furthermore, the level is not elevated in nearly half of patients with the stage I disease, suggesting a limited value as a screening marker. In CA125 positive cases, the marker determination is a sensitive indicator in the early diagnosis of progressive disease in ovarian cancer. The lack of an effective second-line regimen, however, limits the value of the antigen as monitoring marker. Expectations for the new tumor markers lysophosphatidic acid and inhibin are also briefly discussed. PMID- 11915745 TI - [Hygienic control of contingencies associated with local fire events aboard piloted orbital stations]. AB - Based on the multiyear experience of hygienic control of the orbital stations, there are three causes of contingent air pollution: leakage of service system pipeline with chemical working bodies, spills of reagents used in biochemical and technological experiments, and air pollution by products of thermal oxidative degradation of nonmetallic materials. The authors describe their experience in successful elimination and prevention of hazardous consequences of fire aboard the space stations. Analysis of air pollution dynamics after actual fire events in the space station modules showed that it approximates a two-chamber air flow model with two phases of pollutant equilibration: first a rapid fall of concentration in the module on fire till air in the modules gets completely mixed, and then slow elimination of harmful pollutant from air. Identified were markers,--propylene and methylmetacrilate,--reacting to first signs of fire. Carbonic acid nitriles were recognized as toxic products of destruction. The positive effect of intermodular ventilation on dilution of toxic products of thermal oxidative destruction of materials was demonstrated. Satisfactory agreement of calculated and factual pollutant concentrations on board Mir permitted conclusion on applicability of the theoretical model to prediction of levels of air pollution due to contingency. PMID- 11915746 TI - [Development of food rations for humans in extreme environments]. AB - Adequacy of food rations to the human body demands was assessed in three series of investigations with 73 volunteers. The first series (n = 31) was arranged in various climatic-geographic areas (the Arctic, desert, high altitude, and the moderate climate of Russia). Each test continued for 12 days. Energy value of the food ration made up 3560 kilocalories. The second and third series (n = 12 and n = 30, respectively) were performed in a pressurized module with simulation of some extreme factors of space flight. Energy value of the food ration balanced in the content of staple nutrients varied between 2800 and 3500 kcal. Results of physical-hygienic studies suggest adequacy of the proposed food rations to volunteers' physiological demands for nutrients and energy in various extreme conditions. PMID- 11915747 TI - [G-tolerance of female cosmonauts during descent in space flights of 8 up to 169 days in duration]. AB - Investigations into g-tolerance of female cosmonauts during descent in space flights of varying duration are topical for the present day, as there are little data on this issue. Tolerance of +Gz-loads during descent was analyzed based on the data about 4 female cosmonauts in 5 space flights. The space flights were conventionally divided into short- (8-16 days) and long-term (169 days). In two space flights (16 and 169-d long), tubeless anti-g suit Centaur was warn during descent. In these space flights, g-tolerance of females was quite satisfactory advocating for the possibility for women to fly to space without any constraints. When the anti-g suit was not used, female physiological systems were stressed heavier than male. The spacesuit smoothed away this difference. Our results evidence a distinct positive effect of wearing the anti-g suit by female cosmonauts during descent as it reduces stress to their physiological systems. PMID- 11915748 TI - [Mechanisms reducing orthostatic stability of humans in long-term space flight]. AB - Investigations conducted in and following space flights showed consistent decreases in cosmonauts' orthostatic stability due to the effects of long-term microgravity. Degree of this orthostatic disturbance depends on many factors prime of which is adequacy of countermeasures and least consequential is length of space flight, personal features, and others. Shifts in initial circulatory status and establishment of a new level of circulation and functioning of the other body systems in microgravity appear to play the central role in loss of orthostatic stability. The paper contains an in-depth analysis of processes occurring in human body during long-term stay in microgravity impacting orthostatic stability post flight. PMID- 11915749 TI - [A tracking function of human eye in microgravity and during readaptation to earth's gravity]. AB - The paper summarizes results of electro-oculography of all ways of visual tracking: fixative eye movements (saccades), smooth pursuit of linearly, pendulum like and circularly moving point stimuli, pursuit of vertically moving foveoretinal optokinetic stimuli, and presents values of thresholds and amplification coefficients of the optokinetic nystagmus during tracking of linear movement of foveoretinal optokinetic stimuli. Investigations were performed aboard the Salyut and Mir space stations with participation of 31 cosmonauts of whom 27 made long-term (76 up to 438 day) and 4 made short-term (7 to 9 day) missions. It was shown that in space flight the saccadic structure within the tracking reaction does not change; yet, corrective movements (additional microsaccades to achieve tracking) appeared in 47% of observations at the onset and in 76% of observations on months 3 to 6 of space flight. After landing, the structure of vertical saccades was found altered in half the cosmonauts. No matter in or after flight, reverse nystagmus was present along with the gaze nystagmus during static saccades in 22% (7 cosmonauts) of the observations. Amplitude of tracking vertically, diagonally or circularly moving stimuli was significantly reduced as period on mission increased. Early in flight (40% of the cosmonauts) and shortly afterwards (21% of the cosmonauts) the structure of smooth tracking reaction was totally broken up, that is eye followed stimulus with micro- or macrosaccades. The structure of smooth eye tracking recovered on flight days 6-8 and on postflight days 3-4. However, in 46% of the cosmonauts on long-term missions the structure of smooth eye tracking was noted to be disturbed periodically, i.e. smooth tracking was replaced by saccadic. PMID- 11915750 TI - [The model of radiation shielding of the service module of the International space station]. AB - Compared and contrasted were models of radiation shielding of habitable compartments of the basal Mir module that had been used to calculate crew absorbed doses from space radiation. Developed was a model of the ISS Service module radiation shielding. It was stated that there is a good agreement between experimental shielding function and the one calculated from this model. PMID- 11915751 TI - [Phase structure of the ejection period in the human cardiac function before and after 3-day water immersion]. AB - Impedance cardiography was applied to 15 normal male volunteers before and after 3-d water immersion. Determined were parameters of central circulation and dependence of the ejection period on length of the cardiac cycle. Though the ejection period over the whole range of cardiac cycle lengths was shorter than "due" values determined as pre-immersion baseline, cardiac output after immersion was equal to and even slightly above baseline value. According to experimental results, this seems to be linked with little if any dependence of the fast ejection phase, hemodynamically most effective, on heart rate and volume loading which remained same as in the baseline period during all post-immersion tests. In contrast, the reduced ejection period, a variable of the ventricular blood evacuation, is materially dependent on heart rate and volume loading predetermining at large absolute and relative shortening of the ejection period after immersion. PMID- 11915752 TI - [Estimation of the radiation risk of determined effects of human exposure in space]. AB - Subject of the paper is possibility to estimate radiation risk of determined consequences of exposure to solar space rays as a probability of violation of established dose limits. Analysis of specifies of spacecrew exposure to solar space rays in a long-term, particularly interplanetary mission suggests that immediate introduction of the principle in the radiation health policy can result in serious errors, mainly exaggeration, in determination of radiation risk. Proposed are approaches to radiation risk estimation with consideration of the specific of human exposure in space. PMID- 11915753 TI - [The mechanism of inhibition of small intestine contractions following irritation of rabbit's vagus nerve]. AB - Explored was mechanism of inhibition of small intestine contractions due to weak irritation (0.5-2.5 V, 1-30 Hz, 0.3-1.0 ms) of the peripheral segment of rabbit's right vagus nerve. It was shown that the inhibitory effect can be blocked by benzohexamethonium, arphonade, atropine, rhausedyl, and a combination of propranolol and dehydroergotoxine. Inhibition is triggered by catecholamines released by sympathetic endings in consequence of activation of presynaptic M cholinoreceptors on their surface. Presynaptic M-cholinoreceptors are activated by acetylcholine released by the endings of postganglionary parasympathetic nerve fibers. PMID- 11915754 TI - [Activities of certain enzymes in blood of the Pleurodeles waltl newt]. AB - The goal of the study was to evaluate the physiological and biochemical status of Pleurodeles waltli (urodele amphibian) by monitoring enzymatic activity in blood plasma and/or lood cell components. The following enzymes were chosen: glutamate dehydrogenase (GDH), aspartate and alanine aminotransferases (GOT and GPT), superoxide dismutase, catalase, isocitrate dehydrogenase, and glucose-6-phosphate dehydrogenase. With the exception of GDH, GOT and GPT, enzymatic activity was noticeably higher in blood of females as compared to males. Reflecting destructive processes in organism, under normal conditions levels of GOT and GPT activity in plasma are very much equal in females and males. Differences in activities of the other enzymes were proportional to levels of steroid hormones in blood plasma of animals. PMID- 11915755 TI - [Effects of elevated atmospheric ethylene on the development of regenerative organs of short-stem wheat cultivars]. AB - Laboratory experiments were performed to study cytoembryologic development of regenerative organs of wheat cult. 20-1-Shortstem and cult. Apogee in atmosphere with elevated concentration of ethylene. Development of wheat regenerative organs at a concentration of air ethylene of 1 mg/m3 resulted in peculiar changes in floret morphogenesis and partial or complete male barrenness. Cult. Apogee was found to be least vulnerable and, therefore, appears to be suitable for cultivation in greenhouses mounted in pressurized habitats including space station modules. Vulnerability of male reproductive organs in short-stem wheat to ethylene correlates convincingly with the sign of dwarfishness demonstrated by hybrid forms of one of praparents. PMID- 11915756 TI - [Chronic subdural hematoma. Introduction and results of a survey by the French Society of Neurosurgery]. PMID- 11915757 TI - [Chronic subdural hematoma: historical studies]. AB - The first description of a chronic subdural hematoma was made in 1658 by J.J. Wepfer, followed in 1761 by Morgagni. A possible case was described by Honore de Balzac in 1840 including its traumatic origin and surgical treatment. Virchow, in 1857, denied a traumatic origin, and gave the name of "pachymeningitis hemorrhagica interna" to this pathology which he explained by inflammatory processes. The traumatic etiology of chronic subdural hematoma was recognized in the XXth century, especially by Trotter in 1914. Pathophysiology was considered later on in the XXth century. PMID- 11915758 TI - [Physiopathology of chronic subdural hematoma]. AB - Coagulation, fibrinolysis and kinin-kallicrein system disorders could explain chronic subdural hematoma pathophysiology. The marked decrease in antithrombin III in the hematoma suggest that the decreased levels of clotting factors were caused by excessive coagulation resulting in clotting factors consumption. The decrease in alpha 2-antiplasmin with the increase of fibrin degradation products suggest that hyperfibrinolytic activity of the subdural hematoma. Coagulation and fibrinolysis disorders could explain repetitive hemorrhages from the outer membrane, which cause progressive enlargement of the hematoma. Kinin-kallicrein system disorders raise the hemorrhage by its action on the outer membrane. Nowadays, the earliest osmotic theory should reasonably be abandoned. PMID- 11915759 TI - [Clinical forms and prognostic factors of chronic subdural hematoma in the adult]. AB - The most common clinical signs for chronic subdural hematoma in adults are: motor weakness, increased intracranial pressure, confusion and loss of consciousness. This pathology is more frequent in the elderly over 65 years, the diagnosis also being more difficult in this case. Some misleading clinical presentations may delay the decision to perform a CT scan to assess the diagnosis. Preoperative headaches and isodensity in CT-scan are features of good prognosis. Chronic alcoholism and postoperative pneumocephalus are related with poor prognosis. PMID- 11915760 TI - [Chronic subdural hematoma: diagnostic imaging studies]. AB - CT-scan is able to provide the diagnosis of chronic subdural hematoma in more than 90% of the cases. It usually shows a peri-cerebral fluid collection along the convexity, with a convex outer border, and an irregular concave inner border. The density of the collection depends on the age of the hematoma. The main difficulties, in term of diagnosis, result from bilateral isodense chronic subdural hematoma, and differential diagnosis between hematoma, hygroma, and empyema. Some rare localisations can sometimes be seen (posterior fossa, skull base...). A double density with a sedimentation level, or heterogeneity of the hematoma, can sometimes be seen too. Chronic subdural hematoma is demonstrated by MRI in almost 100% of the cases. The intensity of the collection, in T1 and T2 sequences, depends on the age of the hematoma. PMID- 11915761 TI - [Surgical treatment of chronic subdural hematoma in the adult. Review of the literature]. AB - There are various methods to treat chronic subdural hematomas in adults. "Open craniotomy" (craniotomy-membranectomy, burr-hole craniotomy) and "closed craniotomy" (twist-drill craniotomy, subdural tapping) are the most frequent technics, but endoscopy and shunts are also possible. No difference in results can be identified from a review of the literature. Different factors of recurrence are analysed. PMID- 11915762 TI - [Percutaneous treatment of chronic subdural hematoma by twist drill and continuous drainage. Retrospective study of 65 cases]. AB - Twist-drill trephination associated with continuous closed drainage was first proposed forty years ago for treating chronic subdural hematomas. We describe the original percutaneous technique and report our experience in a retrospective study of 65 consecutive patients who have been operated using this method. PMID- 11915763 TI - [Complications of chronic subdural hematoma in the adult]. AB - Chronic subdural hematoma is subject to post-operative fatal and non-fatal complications in 5 to 10% of the cases. Mortality ranges from 0 to 8%, depending on the preoperative clinical status. There is an average recurrence in 8% of the cases, chiefly linked to the absence of drainage. Empyema occurs in 2% of patients, especially when the drain is left in place more than 3 days. In most of the series, long-term epilepsy is a rare complication and patients do not require antiepileptic drugs. The lack of cortical reexpansion, postoperative intracerebral hematoma and tension hydrocephalus are, among others, complications occurring after surgery. Finally, 10% of the patients will have a permanent neurological impairment. PMID- 11915764 TI - [Surgery of idiopathic carpal tunnel syndrome. Comparative study of open techniques and endoscopic techniques (December 2000)]. PMID- 11915765 TI - [Ruptured intracranial aneurysms: endovascular occlusion versus microsurgical exclusion (June 2000)]. PMID- 11915766 TI - Epilepsy Genetics: The 21st Century. Los Angeles, California, USA. December 3, 2000. Proceedings. PMID- 11915768 TI - Guidelines for laparoscopic and conventional surgical treatment of morbid obesity. PMID- 11915767 TI - Proceedings of the 1st International Conference on Colonic Spirochaetal Infections in Animals and Humans. Ekenas, Sweden, April 2-3, 2000. PMID- 11915769 TI - Development of cutaneous sarcoidosis in a patient with chronic hepatitis C treated with interferon alpha 2b. AB - BACKGROUND: Sarcoidosis is a multisystemic granulomatous disorder of unknown etiology that most commonly affects young adults. A probable induction of sarcoidosis by interferons (IFN) has been published. To this date, few cases of cutaneous sarcoidosis inpatients with chronic hepatitis C under interferon treatment have been reported. OBJECTIVE: We describe a 50-year-old woman with chronic hepatitis C who developed lesions of cutaneous sarcoidosis three months after IFN treatment. CONCLUSIONS: The possible role of INF therapy in the development of cutaneous sarcoidosis in a patient with chronic hepatitis C should be considered. PMID- 11915770 TI - [Durability of conventional crowns, bridges and partial prostheses]. PMID- 11915771 TI - [The life span of amalgam restorations]. PMID- 11915772 TI - [Durability of adhesive bridges: a question of survival?]. PMID- 11915773 TI - [Type of restoration in the molars of dentists]. PMID- 11915774 TI - [The activity package of the dental assistant]. AB - The aim of this study was to assess the package of activities that is actually carried out by the chairside assistant, and the activities that should be dropped or added to the package, according to both assistants and dentists. A representative sample of 1034 dentists and their assistants received a questionnaire. Results showed large differences between assistants in kind and number of activities actually carried out. Part of the group of assistants as well as part of the group of dentists expressed their wish to extend the package of activities. PMID- 11915775 TI - [Proceedings of the annual meeting of the American Thoracic Society. San Francisco, California, USA. 18-23 May 2001]. PMID- 11915776 TI - Comparison of carbohydrate and milk-based beverages on muscle damage and glycogen following exercise. AB - This study examined effects of carbohydrate (CHO), milk-based carbohydrate protein (CHO-PRO), or placebo (P) beverages on glycogen resynthesis, muscle damage, inflammation, and muscle function following eccentric resistance exercise. Untrained males performed a cycling exercise to reduce muscle glycogen 12 hours prior to performance of 100 eccentric quadriceps contractions at 120% of 1-RM (day 1) and drank CHO (n = 8), CHO-PRO (n = 9; 5 kcal/kg), or P (n = 9) immediately and 2 hours post-exercise. At 3 hours post-eccentric exercise, serum insulin was four times higher for CHO-PRO and CHO than P (p < .05). Serum creatine kinase (CK) increased for all groups in the 6 hours post-eccentric exercise (p < .01), with the increase tending to be lowest for CHO-PRO (p < .08) during this period. Glycogen was low post-exercise (33+/-3.7 mmol/kg ww), increased 225% at 24 hours, and tripled by 72 hours, with no group differences. The eccentric exercise increased muscle protein breakdown as indicated by urinary 3-methylhistidine and increased IL-6 with no effect of beverage. Quadriceps isokinetic peak torque was depressed similarly for all groups by 24% 24 hours post-exercise and remained 21% lower at 72 hours (p < .01). In summary, there were no influences of any post-exercise beverage on muscle glycogen replacement, inflammation, or muscle function. PMID- 11915777 TI - Three days fast in sportsmen decreases physical work capacity but not strength or perception-reaction time. AB - This study investigates, in young nonobese healthy athletes, the consequences of a 3-day fast coupled, or not, to enhanced physical activity. Eight male subjects, aged 21+/-2 years, fasted for 3 days on two separate occasions, 4 weeks apart. On the first occasion, subjects continued their daily training activities. On the second occasion, a daily physical exercise program was added to these activities. Subjects were evaluated before and after 24 hours and 72 hours of fasting. Evaluation consisted of body composition, basal respiratory exchange ratio, plasma metabolic parameters, perception-reaction time (both simple and discriminant), hand grip strength, and physical work capacity at 170 beats per minute (PWC170). Fasting determined significant reductions in body weight, body fat, and muscle mass. These reductions were not affected by enhanced physical activity. Basal respiratory exchange ratio decreased with fasting but was not influenced by increased training activities. Fasting determined a significant decrease in blood glucose levels, while plasma proteins, urea, uric acid, and free fatty acids increased. Perception-reaction time and hand grip strength were unmodified during fasting. By contrast, PWC170 was significantly and progressively reduced during fasting, and this decrease was not reversed by an increase in training activities. PMID- 11915778 TI - Carbohydrate-loading during the follicular phase of the menstrual cycle: effects on muscle glycogen and exercise performance. AB - The effects of employing a high-carbohydrate diet (carbohydrate-loading) to increase glycogen storage in skeletal muscle are not well established in female athletes. On 4 occasions--2 familiarization trials and 2 experimental trials--6 well-trained female subjects completed 6 x 15-min continuous intervals of cycling (12 min at 72% VO2max, 1 min at maximal effort, and 2 min at 50% VO2max), followed by a time trial 15 min later. The women consumed their habitual diets (HD; 6-7 g carbohydrate/kg lean body mass) for 3 days after the second familiarization trial and before the first experimental trial. During the 3 days following the first experimental trial, the subjects consumed a high-carbohydrate diet (CD; 9-10 g carbohydrate/kg lean body mass) prior to the second experimental trial. Mean (+/-SEM) pre-exercise muscle glycogen concentrations were greater after CD versus HD (171.9+/-8.7 vs. 131.4+/-10.3 mmol/kg wet weight, P < 0.003). Although 4 of the 6 subjects improved their time-trial performance after CD, mean performance for the time trial was not significantly different between diets (HD: 763.9+/-35.6 s; CD: 752.9+/-30.1 s). Thus, female cyclists can increase their muscle glycogen stores after a carbohydrate-loading diet during the follicular phase of the menstrual cycle, but we found no compelling evidence of a dietary effect on performance of a cycling time trial performed after 90 min of moderate intensity exercise. PMID- 11915779 TI - Short-term beta-hydroxy-beta-methylbutyrate supplementation does not reduce symptoms of eccentric muscle damage. AB - PURPOSE: We examined the effects of short-term beta-hydroxy-beta-methylbutyrate (HMB) supplementation on symptoms of muscle damage following an acute bout of eccentric exercise. METHODS: Non-resistance trained subjects were randomly assigned to a HMB supplement group (HMB, 40mg/kg bodyweight/day, n = 8) or placebo group (CON, n = 9). Supplementation commenced 6 days prior to a bout of 24 maximal isokinetic eccentric contractions of the elbow flexors and continued throughout post-testing. Muscle soreness, upper arm girth, and torque measures were assessed pre-exercise, 15 min post-exercise, and 1, 2, 3, 4, 7, and 10 days post-exercise. RESULTS: No pre-test differences between HMB and CON groups were identified, and both performed a similar amount of eccentric work during the main eccentric exercise bout (p > .05). HMB supplementation had no effect on swelling, muscle soreness, or torque following the damaging eccentric exercise bout (p > .05). CONCLUSION: Compared to a placebo condition, short-term supplementation with 40mg/kg bodyweight/day of HMB had no beneficial effect on a range of symptoms associated with eccentric muscle damage. If HMB can produce an ergogenic response, a longer preexercise supplementation period may be necessary. PMID- 11915780 TI - Testosterone responses after resistance exercise in women: influence of regional fat distribution. AB - Regional fat distribution (RFD) has been associated with metabolic derangements in populations with obesity. For example, upper body fat patterning is associated with higher levels of free testosterone (FT) and lower levels of sex-hormone binding globulin (SHBG). We sought to determine the extent to which this relationship was true in a healthy (i.e., non-obese) female population and whether RFD influenced androgen responses to resistance exercise. This study examined the effects of RFD on total testosterone (TT), FT, and SHBG responses to an acute resistance exercise test (ARET) among 47 women (22+/-3 years; 165+/-6 cm; 62+/-8 kg; 25+/-5%BF; 23+/-3 BMI). RFD was characterized by 3 separate indices: waist-to-hip ratio (WHR), ratio of upper arm fat to mid-thigh fat assessed with magnetic resonance imaging (MRI ratio), and ratio of subscapular to triceps ratio (SB/TRi ratio). Skinfolds were measured for the triceps, chest, subscapular, mid-axillary, suprailaic, abdomen, and thigh regions. The ARET consisted of 6 sets of 10 RM squats separated by 2-min rest periods. Blood was obtained pre- and post- ARET. TT, FT, and SHBG concentrations were determined by radioimmunoassay. Subjects were divided into tertiles from the indices of RFD, and statistical analyses were performed by an ANOVA with repeated measures (RFD and exercise as main effects). Significant (p < or = .05) increases following the AHRET were observed for TT (approximately 25%), FT (approximately 25%), and SHBG (4%). With multiple regression analysis, anthropometric measures significantly predicted pre- concentrations of FT, post-concentrations of TT, and pre concentrations of SHBG. The SB/TRi and MRI ratios but not the WHR, were discriminant for hormonal concentrations among the tertiles. In young, healthy women, resistance exercise can induce transient increases in testosterone, and anthropometric markers of adiposity correlate with testosterone concentrations. PMID- 11915781 TI - Prolonged vitamin C supplementation and recovery from demanding exercise. AB - The aim of the present study was to investigate whether 2 weeks of vitamin C supplementation affects recovery from an unaccustomed bout of exercise. Sixteen male subjects were allocated to either a placebo (P; n = 8) or vitamin C group (VC; n = 8). The VC group consumed 200 mg of ascorbic acid twice a day, whereas the P group consumed identical capsules containing 200 mg of lactose. Subjects performed a prolonged (90-min) intermittent shuttle-running test 14 days after supplementation began. Post-exercise serum creatine kinase activities and myoglobin concentrations were unaffected by supplementation. However, vitamin C supplementation had modest beneficial effects on muscle soreness, muscle function, and plasma concentrations of malondialdehyde. Furthermore, although plasma interleukin-6 increased immediately after exercise in both groups, values in the VC group were lower than in the P group 2 hours after exercise (p < .05). These results suggest that prolonged vitamin C supplementation has some modest beneficial effects on recovery from unaccustomed exercise. PMID- 11915782 TI - Osteoporosis knowledge, attitudes, and behaviors of female collegiate athletes. AB - Female athletes often engage in harmful dietary and weight control practices that can impair bone health and hinder performance. To promote related positive health behavior practices, nutrition educators may be more effective if they understand the osteoporosis knowledge, attitudes, and behaviors among female athletes. A questionnaire including items related to osteoporosis and dietary calcium knowledge, attitudes, and behavioral practices was administered to 114 female collegiate athletes (19.6+/-1.4 years). Self-reported intakes of dairy product consumption were also obtained; subjects were asked how many times per week they drank milk and ate cheese, yogurt, and ice cream. The mean score for osteoporosis knowledge was 7.1+/-1.9 (out of 10 items). The mean score for favorable responses to attitude items was 2.1+/-0.8 (out of 3 items). Correct responses to dietary calcium knowledge items were 2.2+/-0.7 (out of 3 items). On average, subjects consumed 2.4+/-1.6 servings of dairy products per day; 31% of subjects consumed the recommended 3 or more servings per day. Osteoporosis knowledge, osteoporosis attitudes, and dietary calcium knowledge were not correlated (p > .05) with dairy product intake. Because of the importance of achieving a high peak bone mass to prevent osteoporosis, our data suggest that further research is needed regarding other factors that might influence dairy product intake among female athletes. PMID- 11915783 TI - Pre-exercise carbohydrate status and immune responses to prolonged cycling: I. Effect on neutrophil degranulation. AB - Ingesting carbohydrate (CHO) beverages during heavy exercise is associated with smaller shifts in numbers of circulating neutrophils and attenuated changes in neutrophil functional responses. The influence of dietary CHO availability on these responses has not been determined. Therefore, the present study investigated the influence of pre-exercise CHO status on circulating neutrophil and lipopolysaccharide (LPS)-stimulated neutrophil degranulation responses to prolonged cycling. Twelve trained male cyclists performed a glycogen-lowering bout of cycling and were randomly assigned to follow a diet ensuring either greater than 70% (HIGH) or less than 10% (LOW) of daily energy intake from CHO for the next 3 days. On day 4, subjects performed an exercise test that comprised cycling for 1 hour at 60% Wmax immediately followed by a time-trial (TT) ensuring an energy expenditure equivalent to cycling for 30 min at 80% Wmax. Subjects repeated the protocol after 7 days, this time following the second diet. The order of the trials was counterbalanced. At TT completion, the HIGH compared with the LOW trial was associated with higher plasma glucose concentration, lower plasma cortisol concentration, and lower circulating neutrophil count. LPS stimulated neutrophil degranulation per cell fell similarly on both trials. These findings suggest that pre-exercise CHO status influences neutrophil trafficking but not function in response to prolonged cycling. PMID- 11915785 TI - Developing a sports nutrition website: an interview with Michelle Minehan. Interview by Louise Burke. PMID- 11915784 TI - Pre-exercise carbohydrate status and immune responses to prolonged cycling: II. Effect on plasma cytokine concentration. AB - Ingesting carbohydrate (CHO) beverages during heavy exercise is associated with smaller changes in the plasma concentrations of several cytokines. The influence of dietary CHO availability on these responses has not been determined. Therefore, the present study investigated the influence of pre-exercise CHO status on plasma interleukin (IL)-6, IL-10, and IL-1 receptor antagonist (IL-1ra) responses to prolonged cycling. Seven trained male cyclists performed a glycogen lowering bout of cycling and were randomly assigned to follow a diet ensuring either greater than 70% (HIGH) or less than 10% (LOW) of daily energy intake from CHO for the next 3 days. On day 4 subjects performed an exercise test that comprised cycling for 1 hour at 60% Wmax immediately followed by a time-trial (TT) ensuring an energy expenditure equivalent to cycling for 30 min at 80% Wmax. Subjects repeated the protocol after 7 days, this time following the second diet. The order of the trials was counterbalanced. At 1 and 2 hours post-TT, plasma concentrations of IL-6 and IL-10 were 2-fold greater on the LOW trial than on the HIGH trial, and peak plasma concentrations of IL-1ra were 9-fold greater on the LOW trial than on the HIGH trial. These findings suggest that pre-exercise CHO status can influence the plasma cytokine response to prolonged cycling. PMID- 11915786 TI - Prophylactic intravenous bolus ephedrine for elective Caesarean section under spinal anaesthesia. AB - BACKGROUND AND OBJECTIVE: To evaluate the efficacy and optimal dose of prophylactic intravenous ephedrine for the prevention of maternal hypotension associated with spinal anaesthesia for Caesarean section. METHODS: After patients had received an intravenous preload of 0.5 L of lactated Ringer's solution, spinal anaesthesia was administered in the sitting position with hyperbaric bupivacaine 2.5 mL 0.5% combined with 25 microg fentanyl. A total of 68 patients were randomized to receive a simultaneous 2 mL bolus intravenously of either 0.9% saline (Group C, n = 20), ephedrine 6 mg (Group E-6, n = 24), or ephedrine 12 mg (Group E-12, n = 22). Further rescue boluses of ephedrine 6 mg were given if systolic arterial pressure fell to below 90 mmHg, greater than 30% below baseline, or if symptoms suggestive of hypotension were reported. RESULTS: There was a significantly higher incidence of hypotension in Group C (60% patients) compared to Group E-12 (27%), but not in Group E-6 (50%). The 95% Confidence Interval for the difference in proportions between Groups C and E-12 was 6-60%, P < 0.05. Fewer rescue boluses of ephedrine were required in Group E-12 compared with Group C (1.8 +/- 1.2 vs. 3.3 +/- 2.1, P < 0.05). There were no significant differences in the incidence of maternal nausea or vomiting, or of neonatal acidaemia between groups. CONCLUSION: A prophylactic bolus of ephedrine 12 mg intravenously given at the time of intrathecal block, plus rescue boluses, leads to a lower incidence of hypotension following spinal anaesthesia for elective Caesarean section compared to intravenous rescue boluses alone. PMID- 11915787 TI - Bronchial obstruction treated by fibreoptic-guided suction. PMID- 11915788 TI - Review: moderately early postnatal corticosteroids reduce chronic lung disease and mortality in preterm infants, but increase complications. PMID- 11915789 TI - Review: self help, media-based therapy is effective for behaviour problems in children. PMID- 11915790 TI - Review: family and parenting interventions reduce subsequent arrests and length of time in institutions in youths with conduct disorder and delinquency. PMID- 11915791 TI - Review: albumin administration is not associated with excess mortality in acutely ill patients. PMID- 11915792 TI - Review: double bag or Y-set systems reduce peritonitis in patients on continuous ambulatory peritoneal dialysis. PMID- 11915793 TI - Total contact casts were better than removable cast walkers or half shoes for healing diabetic neuropathic foot ulcers. PMID- 11915794 TI - Listening to music during ambulatory ophthalmic surgery reduced blood pressure, heart rate, and perceived stress. PMID- 11915795 TI - An audiotaped information programme after coronary artery bypass surgery improved physical functioning in women and psychological distress in men. PMID- 11915797 TI - An outpatient geriatric evaluation and management programme was more effective than usual care in preventing functional decline in high risk older adults. PMID- 11915796 TI - Review: specialised multidisciplinary follow up reduces hospital admissions but not mortality in patients with heart failure. PMID- 11915798 TI - Review: pharmacological and non-pharmacological interventions improve outcomes in patients with dementia and their caregivers. PMID- 11915799 TI - Training family care givers of people with dementia to think in a more clinical manner decreased depression and the sense of burden. PMID- 11915800 TI - A home based, nurse delivered exercise programme reduced falls and serious injuries in people >/= 80 years of age. PMID- 11915801 TI - Review: hip protectors reduce hip fractures after falls in elderly people living in institutions or supported home environments. PMID- 11915803 TI - Progression of disease in HIV infected children slowed after the first year of life. PMID- 11915802 TI - Preoperative pillow placement under the injured extremity had better analgesic effects than skin traction for hip fracture. PMID- 11915804 TI - Interactive continuing education workshops or conferences can improve professional practice and patient outcomes. PMID- 11915805 TI - Low income mothers of overweight children had personal and environmental challenges in preventing and managing obesity. PMID- 11915806 TI - Parents' perceptions of obtaining a diagnosis of childhood cancer can include experiences of disputes and delays. PMID- 11915808 TI - Patient taciturnity in health counselling was understood in terms of 4 participation frames. PMID- 11915807 TI - Emergency department care of women who were abused was driven by the prevailing practice pattern of efficient patient processing. PMID- 11915809 TI - Attitudes about organ transplantation reflected different conceptions of the body. PMID- 11915810 TI - Spouses who placed partners in care homes experienced emotional reactions to separation and made efforts to maintain their relationship. PMID- 11915811 TI - Print and online versions of Evidence-Based Nursing: innovative teaching tools for nurse educators. PMID- 11915812 TI - Review: early postnatal corticosteroids reduce chronic lung disease in preterm infants, but increase complications. PMID- 11915814 TI - Risperidone was effective for aggression in adolescents with disruptive behaviour disorders and below average intelligence. PMID- 11915813 TI - Lithium is more effective than placebo for preventing all relapses in patients with bipolar but not unipolar disorder. PMID- 11915815 TI - Life skills training and booster sessions plus drinking reduction treatment was effective in heavily drinking women. PMID- 11915816 TI - Review: eye movement desensitization and reprocessing is not better than exposure therapies for anxiety or trauma. PMID- 11915817 TI - Brief psychodynamic interpersonal therapy after deliberate self poisoning reduced suicidal ideation and deliberate self harm. PMID- 11915818 TI - Review: placebo is better than no treatment for subjective continuous outcomes and for treatment of pain. PMID- 11915820 TI - Review: iron treatment does not improve psychomotor development and cognitive function at 30 days in children with iron deficiency anaemia. PMID- 11915819 TI - Paroxetine but not imipramine improved depression related outcomes in adolescents with DSM-IV major depression. PMID- 11915821 TI - Olanzapine was more effective than lorazepam at 2 hours but not at 24 hours in bipolar mania with acute agitation. PMID- 11915822 TI - Ongoing contact for 2 years after discharge was associated with greater survival in patients at risk of post crisis suicide. PMID- 11915823 TI - Once a day Concerta methylphenidate was equivalent to 3 times daily methylphenidate in children with ADHD. PMID- 11915825 TI - Repeated doses of porcine secretin did not improve symptoms, language, or cognitive functioning in children with autism or autism spectrum disorder. PMID- 11915824 TI - Quintuple the daily maintenance dose of buprenorphine given every 5 days was associated with opioid withdrawal symptoms. PMID- 11915826 TI - Cognitive treatment reduced symptoms and improved functioning in bipolar disorder. PMID- 11915827 TI - St John's wort was not better than placebo for reducing depression scores. PMID- 11915828 TI - Paroxetine and sertraline delayed ejaculation in heterosexual men with lifelong rapid ejaculation. PMID- 11915829 TI - Review: most laboratory tests do not add to the diagnostic accuracy of clinical criteria for dementia. PMID- 11915830 TI - A risk assessment system based on preoperative factors was fairly specific in predicting postoperative delirium risk in patients having major elective surgery. PMID- 11915831 TI - A 4 stage model described how clients and their relatives experienced the process of developing schizophrenia. PMID- 11915832 TI - Parents' experience of having children with a significant developmental disability. PMID- 11915833 TI - Drug users' experiences of injecting included descriptions of needle fixation. PMID- 11915835 TI - Caregivers described how an Alzheimer's disease respite programme gave them time to attend to their own needs. PMID- 11915834 TI - General practitioners often used emotional responses for "referrals away" and intellectual decision making for "referral to" in patients with mild depression and/or anxiety. PMID- 11915836 TI - A geriatric evaluation and management programme prevented functional decline and reduced depression in high risk older adults. PMID- 11915837 TI - Proceedings of the 4th Joint Meeting of Italian-Hungarian Pancreatologists. Capri, Italy, September 30, 2000. PMID- 11915838 TI - Cystoid macular edema in the era of ocular hypotensive lipids. AB - PURPOSE: To review cystoid macular edema in relationship with ocular hypotensive lipids and to present recommendations for use in eyes at increased risk of cystoid macular edema. DESIGN: Focused literature review and authors' clinical experience. METHODS: Cystoid macular edema in association with cataract surgery, epinephrine therapy, and ocular hypotensive lipid therapy is reviewed. RESULTS: Cystoid macular edema may be associated with a variety of therapeutic interventions, predominantly in eyes at increased risk with open or absent posterior lens capsule and ocular inflammation. Based primarily on experience with latanoprost, the association between ocular hypotensive lipids and cystoid macular edema, even in high-risk eyes, although present, appears to be low. CONCLUSIONS: Considering the low incidence of cystoid macular edema associated with latanoprost therapy, the authors feel that after maximal tolerable medical therapy and with proper precautions, the use of an ocular hypotensive lipid is an acceptable alternative to filtration surgery in high-risk eyes. PMID- 11915839 TI - Insights on Infection and Immunity: A Festschrift in Honor of Jack Remington. Palo Alto, California, USA, 19 January 2001. PMID- 11915840 TI - Thinking about women's health: the case for gender sensitivity. PMID- 11915841 TI - Abstracts from the 13th EUROSON Congress, 33rd BMUS Annual Meeting. Edinburgh, Scotland, 11-14 December 2001. PMID- 11915842 TI - Re: Feild TS, et al. Why leaves turn red in autumn. The role of anthocyanins in senescing leaves of red-osier dogwood. [2001] Plant Physiol 127:566-574. PMID- 11915843 TI - Abscisic acid: a universal signalling factor? PMID- 11915844 TI - The role of patients' meta-preferences in the design and evaluation of decision support systems. AB - The arrival of new analysis-based decision technologies will necessitate a profound rethinking both of the nature of the patient-doctor relationship and of the way aids and support systems designed to improve decision-making within that relationship are designed and evaluated. One-dimensional typologies of the traditional 'paternalist/shared/informed' sort do not provide the complexity called for by the heterogeneity of patient's 'meta-preferences' regarding their relationship with a doctor on the one hand and regarding the analytical level of judgement and decision-making on the other. A multidimensional matrix embodying this distinction is proposed as a framework of the minimal complexity required for the design and evaluation of the full range of decision aids and decision modes. Essentially aids should be conceived of and evaluated cell-specifically and the search for universally satisfactory decision support systems abandoned. 'shared' and'informed' are best interpreted as attributes which may or not be in line with a patient's meta-preferences. Future research should focus on the higher level goal of better decision-making, a goal that will need to respect and reflect these meta-preferences. PMID- 11915845 TI - Whose priorities count? Comparison of community-identified health problems and Burden-of-Disease-assessed health priorities in a district in Uganda. AB - OBJECTIVES: The aim of the study was to compare health problems as defined quantitatively by the Burden of Disease study to those defined by the community. The secondary aim was to explore the potential for using qualitative participatory methodologies as tools for planing and priority setting. DESIGN: Interviews and group discussions with a purposely sampled set of community members (n = 51) and community leaders(n = 6). The Nominal group technique, as well as in-depth interviews, were used to identify major health problems - as perceived by the community. Epidemiological data on the major health problems were derived from the national Burden of Disease study. RESULTS: Community perceived health problems were similar to those identified by the burden of disease study. Reasons given for the ranking included prevalence, fatality, social and cultural stigma. Social stigma and cultural values were not considered in the burden of disease studies. However, socially stigmatized diseases were considered to be more serious compared to non-stigmatised conditions, in spite of their low prevalence. Poverty and lack of knowledge were the perceived major causes of ill-health in the community. CONCLUSIONS: Qualitative approaches like the nominal group technique may be useful in eliciting community values that could supplement quantitative information like that elicited by the Burden of Disease study. Such a mixed approach would capture both epidemiologicaly assessed and community felt needs in the priority setting process. PMID- 11915846 TI - Chromatin and transcription: where do we go from here. PMID- 11915847 TI - Cardiovascular reactivity in Black South-African males of different age groups: the influence of urbanization. AB - OBJECTIVE: Blacks in an urban environment seem to be more vulnerable to excessive increases in blood pressure during daily life events. This greater cardiovascular reactivity during acute stress situations in urban Blacks may lead to the development of hypertension in their later lives. Because Blacks in South Africa are involved in a massive process of urbanization, which may lead to chronic diseases of lifestyle like hypertension and diabetes, this study was undertaken to compare the cardiovascular reactivity patterns of urbanized and rural Black males in the North-West province of South Africa. DESIGN: Two hundred twenty three Black males of different age groups, Group 1: younger than 25 years of age; Group 2: between 25 and 44 years of age; and Group 3: 45 years of age and older. Participants were randomly selected from rural and urbanized settlements. METHODS: After resting blood pressure was recorded with a Finapres apparatus, and cardiac output, stroke volume, heart rate, and total peripheral vascular resistance had been obtained, an acute laboratory stressor (hand dynamometer exercise) was applied and the above measurements repeated. RESULTS: The SBP, DBP, and MAP increased 28% and 42% in both the rural and the urbanized groups. There was a shift from a central reactivity pattern (increased cardiac output) in the young rural group (< 25 years) to a peripheral reactivity pattern (increased total peripheral resistance) in the > or = 45 years old urbanized males after applying the stressor. CONCLUSIONS: Cardiovascular reactivity differs between Black males from a rural area compared to urbanized Black males. The urbanized males > or = 45 years of age were at a higher risk of developing cardiovascular disease because their total peripheral resistance reactivity increased the most during application of the stressor. PMID- 11915848 TI - Treatment of heart failure in patients with diabetes: clinical update. AB - Heart failure is an important problem for the growing number of persons afflicted with diabetes. Not only is HF more prevalent than in the non-diabetic population but also it carries a graver prognosis. Diabetic cardiomyopathy adds to the already increased risks for developing HF with hypertension and coronary heart disease, leading to systolic and diastolic dysfunction. As LV function deteriorates, increases in RAS and SNS activation may lead to progressive HF and premature death. The greater prevalence and increased mortality risk in diabetic patients supports the role of aggressive HF diagnosis and management, even in asymptomatic patients. Treatment strategies already established for HF have been shown to be effective in the diabetic population, including ACE inhibitors and beta-blockers. There is evidence that glycemic control is also important in improving clinical outcomes. The adverse effects of beta-blockade on carbohydrate and lipid metabolism and vasoconstriction may be circumvented by the use of carvedilol. PMID- 11915849 TI - Prevalence of lower extremity arterial disease defined by the ankle-brachial index among American Indians: the Inter-Tribal Heart Project. AB - OBJECTIVES: This study examines the prevalence of lower extremity arterial disease (LEAD) and its association with cardiovascular disease (CVD) risk factors. METHODS: Linear and logistic regression were used to analyze cross sectional data from 1333 Chippewa and Menominee Indians. RESULTS: Approximately 6.4% of participants (6.9% of men, 6.1% of women) had LEAD. Among women, LEAD was associated with lower high-density lipoprotein cholesterol; higher levels of diastolic blood pressure, creatinine, and triglycerides; and current smoking, diabetes, microalbuminuria, prior myocardial infarction, and stroke (P < or = .05). Among men, LEAD was associated with higher levels of creatinine (P < or = .05). CONCLUSIONS: Use of the ankle-brachial index to assess LEAD is a noninvasive and inexpensive measurement that can be used by clinicians to identify persons at increased risk of developing CVD, and may provide opportunities to prevent CVD. PMID- 11915851 TI - Convergent maternal care strategies in ungulates and macropods. AB - Mammals show extensive interspecific variation in the form of maternal care. Among ungulates, there is a dichotomy between species in which offspring follow the mother ("following" strategy) versus species in which offspring remain concealed ("hiding" strategy). Here we reveal that the same dichotomy exists among macropods (kangaroos, wallabies and allies). We test three traditional adaptive explanations and one new life history hypothesis, and find very similar patterns among both ungulates and macropods. The three traditional explanations that we tested were that a "following" strategy is associated with (1) open habitat, (2) large mothers, and (3) gregariousness. Our new life-history hypothesis is that a "following strategy" is associated with delayed weaning, and thus with the "slow" end of the slow-fast mammalian life-history continuum, because offspring devote resources to locomotion rather than rapid growth. Our comparative test strongly supports the habitat structure hypothesis and provides some support for this new delayed weaning hypothesis for both ungulates and macropods. We propose that sedentary young in closed habitats benefit energetically by having milk brought to them. In open habitats, predation pressure will select against hiding. Followers will suffer slower growth to independence. Taken together, therefore, our results provide the first quantitative evidence that macropods and ungulates are convergent with respect to interspecific variation in maternal care strategy. In both clades, differences between species in the form of parental care are due to a similar interaction between habitat, social behavior, and life history. PMID- 11915850 TI - Serologic evidence of ehrlichiosis among humans and wild animals in The Netherlands. AB - The seroprevalence of antibodies directed against granulocytic and monocytic Ehrlichia was determined by use of human granulocytic ehrlichiosis agent and Ehrlichia chaffeensis as surrogate antigens. Seven hundred twenty-one serum samples were collected between 1992 and 1999 from febrile patients with unresolved aetiology (n=108), patients suspected of having Lyme disease (n=174), forestry workers (n=154) and healthy controls (n=54) as well as from wild deer (n=96), hares (n=60), wild boar (n=15) and red foxes (n=60). Reactive antibodies against granulocytic Ehrlichia were detected in 4% of febrile patients with unresolved aetiology and in 4% of patients suspected of having Lyme disease. Among the forestry workers, 1% tested positive for antibodies against granulocytic Ehrlichia, whereas all the healthy controls were negative. Antibody reaction against monocytic Ehrlichia was detected in only 2% of the febrile patients. Granulocytic Ehrlichia and monocytic Ehrlichia-reactive serum antibodies were detected in 22% and 3% of the deer samples, respectively, and in 2% of the hares. In wild boars and in red foxes, only serum antibodies reactive against monocytic Ehrlichia were detected in 13% and 7%, respectively. The demonstration of the presence of both granulocytic and monocytic Ehrlichia reactive serum antibodies among humans and wild animals in The Netherlands indicates that patients suspected of having Lyme disease and febrile patients with unresolved aetiology should be tested for the presence of granulocytic and monocytic Ehrlichia antibodies or by polymerase chain reaction. Furthermore, granulocytic Ehrlichia are most prevalent in humans and animals in The Netherlands. PMID- 11915852 TI - The evolution and maintenance of monoecy and dioecy in Sagittaria latifolia (Alismataceae). AB - Plant species rarely exhibit both monoecious and dioecious sexual systems. This limits opportunities to investigate the consequences of combined versus separate sex function on mating patterns and genetic variation and the analysis of factors responsible for the evolution and maintenance of the two sexual systems. Populations of the North American clonal aquatic Sagittaria latifolia are usually either monoecious or dioecious and often grow in close geographic proximity. We investigated mating patterns, genetic structure, and relationships between the two sexual systems using allozyme variation in populations from southern Ontario, Canada. As predicted, selfing rates in monoecious populations (n = 6, mean = 0.41) were significantly higher than in dioecious populations (n = 6, mean = 0.11). Moreover, marker-based estimates of inbreeding depression (delta) indicated strong selection against inbred offspring in both monoecious (mean delta = 0.83) and dioecious (mean delta = 0.84) populations. However, the difference in selfing rate between the sexual systems was not reflected in contrasting levels of genetic variation. Our surveys of 12 loci in 15 monoecious and 11 dioecious populations revealed no significant differences in the proportion of polymorphic loci (P), number of alleles per locus (A), or observed and expected heterozygosity (H(o) and H(e), respectively). Strong inbreeding depression favoring survival of outcrossed offspring may act to maintain similar levels of diversity between monoecious and dioecious populations. Despite geographical overlap between the two sexual systems in southern Ontario, a dendrogram of genetic relationships indicated two distinct clusters of populations largely corresponding to monoecious and dioecious populations. Reproductive isolation between monoecious and dioecious populations appears to be governed, in part, by observed differences in habitat and life history. We suggest that selfing and inbreeding depression in monoecious populations are important in the transition from monoecy to dioecy and that the maintenance of distinct sexual systems in S. latifolia is governed by interactions between ecology, life history, and mating. PMID- 11915853 TI - Population and colony genetic structure of the primitive termite Mastotermes darwiniensis. AB - The termite Mastotermes darwiniensis is the sole extant member of its family and occupies the basal position in the phylogeny of the eusocial order Isoptera. In this study, we investigated the micro- and macrogeographic genetic structure of M. darwiniensis in its native range in Australia. A total of 1591 workers were sampled from 136 infested trees in 24 locales. Each locale was separated by 2-350 km, and these locales were found within two broader geographic regions approximately 1500 km apart. The multilocus genotypes of all termites were assayed at six polymorphic microsatellite loci. The genetic data indicated that colonies typically fed on multiple trees within locales and extended over linear distances of up to 320 m. Single colonies were frequently headed by multiple reproductives. Workers were highly related (r = 0.40) and substantially inbred (f = 0.10). Thus, M. darwiniensis colonies are characterized by the input of alleles from multiple reproductives, which sometimes engage in consanguineous matings. Our analyses of population genetic structure above the level of the colony indicated that locales and regions were significantly differentiated (theta(locale) = 0.50, theta(region) = 0.37). Moreover, locales showed a pattern of genetic isolation by distance within regions. Thus, M. darwiniensis populations display restricted gene flow over moderate geographic distances. We suggest that the genetic patterns displayed by M. darwiniensis result primarily from selective pressures acting to maintain high relatedness among colonymates while allowing colonies to grow rapidly and dominate local habitats. PMID- 11915855 TI - The International Menopause Society menopause-related terminology definitions. PMID- 11915854 TI - Comparison of Alora estradiol matrix transdermal delivery system with oral conjugated equine estrogen therapy in relieving menopausal symptoms. Alora Study Group. AB - OBJECTIVE: To compare the efficacy of two strengths of an estradiol matrix transdermal delivery system with daily oral doses of conjugated equine estrogens in reducing the frequency of moderate-to-severe hot flushes in postmenopausal women. DESIGN: The design of the study provided for the following treatment regimens: an estradiol transdermal delivery system (Alora 0.05 or 0.1 mg/day) administered twice weekly or oral doses of conjugated equine estrogens (CEE 0.625 or 1.25 mg) administered daily were given to 321 highly symptomatic postmenopausal women for 12 weeks following a randomized, parallel-group, double blind, double-dummy design. RESULTS: Results indicate no statistically significant differences at any time point in mean frequency or mean percentage reduction in frequency of moderate-to-severe hot flushes between patients given Alora 0.1 mg/day and those receiving CEE 1.25 mg/day. Similarly, no significant differences were observed at any time in mean frequency of moderate-to-severe hot flushes between the Alora 0.05 mg/day and CEE 0.625 mg/day groups, although the group receiving CEE 0.625 mg/day exhibited a statistically greater percentage reduction than the Alora 0.05 mg/day group at weeks 3, 4 and 8. By week 12, these two treatments were statistically indistinguishable. There were no serious or unexpected adverse events with the two transdermal systems and local skin tolerability was excellent. Other estrogenic effects such as restoration of vaginal cytology, breast tenderness and unexpected vaginal bleeding were comparable between transdermal and oral administration groups except for a lower incidence of bleeding in those women receiving the lower dose transdermal regimen. PMID- 11915856 TI - Psychometric properties of the Chinese version of the Family Awareness Scale. AB - The author examined the reliability and validity of the Chinese Family Awareness Scale (C-FAS) in 2 studies. In Study 1, data from 361 adolescents (a) showed that the C-FAS was temporally stable and internally consistent and (b) supported the concurrent and construct validities of the scale. In Study 2, data from clinical (N = 281) and nonclinical (N = 451) groups of adolescents showed that the C-FAS scores discriminated between the 2 groups. C-FAS scores were also correlated significantly with other measures of family functioning and psychological well being. Results of the 2 studies suggest that the C-FAS displayed generally good psychometric properties in different samples of Chinese adolescents. PMID- 11915857 TI - Acceptability of personal hormone monitoring for contraception: longitudinal and contextual variables. AB - A new contraceptive technology may advance the science of family planning but may do little to affect health if potential users do not deem it an acceptable method. The authors conducted an acceptability study of a newly developed contraceptive method--personal hormone monitoring. A sample of 480 English volunteers present at the 6th month of a 13-month longitudinal study completed surveys regarding their attitudes toward a personal hormone monitor for the purpose of contraception. The authors used the participants' responses to determine (a) the extent to which the participants accepted the monitor, (b) how their ratings of acceptability changed over time, (c) the extent to which contextual variables predicted changes in acceptability over time, and (d) whether those contextual variables predicted final acceptability of the monitor. Results suggested that no single method of family planning is best for everyone and specified the people for whom personal hormone monitoring may be most suitable. PMID- 11915859 TI - Abstracts of the American Urological Association Annual Meeting. May 25-30, 2002. Orlando, Florida, USA. PMID- 11915858 TI - The performance of four different corneal topographers on normal human corneas and its impact on orthokeratology lens fitting. AB - PURPOSE: To evaluate the performances of Humphrey Atlas 991, Orbscan II, Dicon CT200, Medmont E300 on young Chinese adults. METHODS: Three sets of corneal topography measurements were obtained from each topographer from 22 subjects-two sets by the same examiner and one set by another examiner on the same day. RESULTS: There were no significant within-examiner and between-examiner differences for any of the parameters tested for each topographer. However, only the repeatability and reproducibility (of apical radius[Ro], eccentricity, and elevation) of the Humphrey and Medmont were good. There was no statistically significant between-topographer difference in R(o), but significant differences in eccentricity and elevation values were found. The number of repeated readings that should be taken for a precision of 2 microm (elevation) were 12 for the Humphrey and 2 for the Medmont. CONCLUSIONS: The performance of both the Humphrey and the Medmont was very good. R(o) and eccentricity values of different topographers cannot be used interchangeably, but the agreement in elevation values was good for these topographers. The number of repeated readings required for maximum precision varies with the topographer used, and they are not interchangeable. PMID- 11915860 TI - SCID mouse models to study human cancer pathogenesis and approaches to therapy: potential, limitations, and future directions. AB - The successful engraftment of human tumors and human immunocompetent cells into severe combined immunodeficient (SCID) mice has led to the generation of a wide array of different experimental designs that have proven useful in studying the cell biology of human cancer, and for evaluating novel therapeutic approaches to the treatment of cancer. In this review five of the most frequently used embodiments of the SCID model are presented. The goals of this review are to discuss how each model has been utilized to study human cancer and its response to many different novel therapies, to provide an assessment of the strengths and limitations of each model, and to outline future directions with a focus on what is needed to overcome some of the current limitations and pitfalls of the SCID models. PMID- 11915861 TI - Two patients with microscopic polyangiitis and unusual pulmonary manifestation. AB - We encountered two patients with microscopic polyangiitis (MPA) associated with unusual pulmonary manifestations. The first patient was a 45-year-old man who had worked in amine for 3 years when he was young. On admission, chest X-rays showed long-standing silicosis and a new patchy infiltration. The second patient was a 52-year-old female. On admission, chest X-rays showed bilateral patchy infiltrations. Since then, variable patterns of patchy infiltration have waxed and waned repeatedly. The renal biopsy revealed that both patients had glomerulonephritis associated with small vessel vasculitis but with few or no immune deposits. There was neither granulomatous inflammation nor eosinophilic infiltration. Myeloperoxidase-antineutrophil cytoplasmic antibody (ANCA) was positive in both patients. After treatment with glucocorticoids and cyclophosphamide, radiological findings were minimal and stable. These two cases show that patients with MPA have a wide spectrum of radiological findings. PMID- 11915862 TI - Pulmonary nocardiosis in a patient treated with corticosteroid therapy. AB - We report a case of pulmonary nocardiosis in a 69-year-old man with rheumatoid arthritis who was receiving corticosteroid treatment. The patient received prednisolone for rheumatoid arthritis and antibiotics for his fever and pneumonia in another hospital, but the response to the therapy was poor. After admission to our hospital, he improved following treatment with imipenem/cilastatin for Nocardia asteroides. Pulmonary nocardiosis is difficult to diagnose and should be considered in the differential diagnosis, especially in an immunocompromised host. PMID- 11915863 TI - Voluntary health insurance in the European Union: a critical assessment. AB - The authors examine the role and nature of the market for voluntary health insurance in the European Union and review the impact of public policy, at both the national and E.U. levels, on the development of this market in recent years. The conceptual framework, based on a model of industrial analysis, allows a wide range of policy questions regarding market structure, conduct, and performance. By analyzing these three aspects of the market for voluntary health insurance, the authors are also able to raise questions about the equity and efficiency of voluntary health insurance as a means of funding health care in the European Union. The analysis suggests that the market for voluntary health insurance in the European Union suffers from significant information failures that seriously limit its potential for competition or efficiency and also reduce equity. Substantial deregulation of the E.U. market for voluntary health insurance has stripped regulatory bodies of their power to protect consumers and poses interesting challenges for national regulators, particularly if the market is to expand in the future. In a deregulated environment, it is questionable whether this method of funding health care will encourage a more efficient and equitable allocation of resources. PMID- 11915865 TI - Cumulative combined subject and author index, volumes 1-6, 1985-2001. PMID- 11915864 TI - Ruminal supplementation of direct-fed microbials on diurnal pH variation and in situ digestion in dairy cattle. AB - To evaluate the effect of direct-fed microbial (DFM) concentration on diurnal rumen pH profiles and in situ digestibilities, nine ruminally cannulated cows in early lactation were fed treatments consisting of DFM (Enterococcus faecium, Lactobacillus plantarum, and Saccharomyces cerevisiae) at a level of a) 1 x 10(5) cfu/ml of rumen fluid (10(5)), b) 1 x 10(6) cfu/ml of rumen fluid (10(6)), and c) 1 x 10(7) cfu/ml rumen fluid (10(7)). Treatments were directly administered via rumen cannula once daily. Cows were fitted with pH probes in their cannula and connected to dataloggers, which monitor pH hourly. The experimental period was 21 d: 7-d adjustment, 14-d for pH, and in situ measurements. Cows fed 10(5) were able to sustain a higher nadir pH than were cows fed 10(6) or 10(7). Cows fed 10(5) had a higher digestion rate of high moisture ear corn (HMEC) dry matter. Corn silage digestion was higher for cows fed 10(5) and 10(6) compared with those receiving 10(7). There were no carryover effects of treatment associated with rumen pH when switching from one treatment regimen to the next. Results from this study demonstrate that incorporation of a specific level of DFM aids in reducing diurnal ruminal acidity. PMID- 11915866 TI - Protocol for assessing brain function in fish and the effectiveness of methods used to stun and kill them. AB - A method for the evaluation of brain function in fish has been developed which is based on assessments of self-initiated behaviours, responses to stimulation, and reflexes. These assessments were validated in several freshwater and marine species and applied to evaluate the brain function of fish while they were anaesthetised and while they were being killed by a variety of methods. The results of these investigations were compared with published neurophysiological observations and it is concluded that some of the tests can be used to identify, with reasonable confidence, the state of awareness, and thus the capacity to experience suffering of several species of fish. PMID- 11915867 TI - Proceedings of the Physiological Society Scientific Meeting held at University of York, 17-19 December 2001. PMID- 11915874 TI - The Royal College of Ophthalmologists cataract surgery guidelines: what can patients see with their operated eye during cataract surgery? PMID- 11915873 TI - A sampling and analytical methodology for dental trace element analysis. AB - The role of trace elements in human health and environmental pollution has developed into an extensive field of research. This study describes a sampling and analytical strategy to determine the trace element content of primary (deciduous) teeth and to assess their use in environmental health and nutrition studies. Exfoliated and extracted primary teeth were collected from 21 Ugandan and 27 UK children. The crown and root of the teeth were separated and the former digested and analysed for several elements by inductively coupled plasma mass spectrometry. The influence of country, tooth type, age and gender were statistically investigated in addition to within-person variation. A principal components analysis (PCA) was used to treat the data in a multivariate fashion and facilitated the moderation of outliers. The results demonstrated that country of origin has an important influence on the elemental composition of teeth and that tooth type should be controlled in these types of studies. Given such a restriction, the age and gender of the donor should have no effect and do not need to be controlled. In addition, where country of domicile, age and gender were controlled, the concentrations of most elements within a single tooth type were representative of an individual and therefore may be indicative of health status. PMID- 11915875 TI - Pterygium excision and conjunctival mini-autograft. PMID- 11915876 TI - Non-trachomatous corneal opacities in the Gambia--aetiology and visual burden. AB - AIMS: National blindness surveys conducted in the Gambia in 1986 and 1996 showed an increase in blindness and visual impairment from non-trachomatous opacity. This study aimed to investigate the aetiology of these opacities and to assess the resulting visual burden. METHODS: A population-based, randomised blindness survey was conducted in the Gambia in 1996. Patients with visual impairment or blindness were examined by an ophthalmologist with a slit lamp. Causes of corneal opacity were determined as accurately as possible by clinical history and examination. RESULTS: A total of 154 patients with non trachomatous corneal opacity were examined of whom 39 had bilateral opacities and 115, unilateral. Causes included corneal infection, measles/vitamin A deficiency, harmful traditional practices and trauma (unilateral scarring). Overall, corneal pathology alone was responsible for bilateral visual impairment or blindness in 19 (12%) patients and unilateral visual impairment or blindness in 88 (57%) patients. Those patients with bilateral visual impairment or blindness (mean age 59, SD) were older (P= 0.003) than others (mean age 44, SD = 20). The use of harmful traditional eye practices was associated with bilateral corneal blindness or visual impairment (RR = 2.63, 95% CI 1.11-6.21, P = 0.04). Although none of the corneal scars reported here were attributed to trachoma, in patients over the age of 45, the prevalence of trachomatous conjunctival scarring in this group was 38.8% compared to 19.4% of the whole nationwide sample. DISCUSSION: Strategies for the prevention (including the quest for cheaper anti-microbial drugs and co operation with traditional healers) and surgical treatment of these corneal opacities are discussed. PMID- 11915877 TI - Tumor Prevention and Genetics. Abstracts of the 2nd International Conference and 7th Annual Meeting of the International Society of Cancer Chemoprevention. February 2002, St Gallen, Switzerland. PMID- 11915878 TI - European Academy of Andrology membership list 2001, Andrology Centers. PMID- 11915879 TI - Acute, oedematous Mycobacterium ulcerans infection in a farmer from far north Queensland. PMID- 11915880 TI - Comparison of the microbial composition of voice prosthesis biofilms from patients requiring frequent versus infrequent replacement. AB - This study was performed to establish a possible difference in biofilm composition in patients who require frequent versus infrequent prosthesis replacement. Only Groningen button voice prostheses that were removed because of increased airflow resistance or leakage of food or liquids through the prosthesis were considered for this study. These prostheses were selected from a total of 692 failed voice prostheses over a 2-year evaluation period. The failed voice prostheses were subdivided into a short-lifetime group, corresponding to an implantation period of less than 4 months (20 voice prostheses), and an extended lifetime group, corresponding to an implantation period of greater than 9 months (18 voice prostheses). The biofilm was removed from the valve sides of the prostheses. The bacterial strain Rothia dentocariosa and the yeast strains Candida albicans I and Candida tropicalis were the predominant strains isolated from the biofilms on the voice prostheses in the short-lifetime group, whereas in the extended-lifetime group, R dentocariosa was found with a fourfold lower isolation frequency and C albicans I was found with a twofold lower isolation frequency. Candida tropicalis was absent from the extended-lifetime group. PMID- 11915882 TI - Physiological effects of gravitation. AB - The physiological effects of weightlessness are discussed, as they were measured and reported by the Russian cosmonauts. The possibilities of adaptation to space environment by training and the mechanisms of compensation and vicarization (adaptive substitution) of the analyzers and processes of the biological entity are considered. The authors present recommendations for cosmonaut training for distant and prolonged flights. PMID- 11915881 TI - Longitudinal and cross-sectional phenotype analysis in a new, large Dutch DFNA2/KCNQ4 family. AB - We analyzed hearing thresholds, speech recognition scores, and vestibular responses in 32 affected persons in a large family with DFNA2/KCNQ4-related hearing impairment caused by a W276S missense mutation. Linear regression analysis of individual longitudinal data revealed significant threshold progression (1 dB/y) and offset (at age zero). The mean offset thresholds were 5, 21, 40, 39, 31, and 51 dB hearing level (HL) at 0.25, 0.5, 1, 2, 4, and 8 kHz, respectively. Cross-sectional analysis of last-visit thresholds against age produced less-steep slopes and higher offset thresholds. Nonlinear regression analysis of last-visit phoneme recognition scores against age in 25 cases showed that speech recognition did not deteriorate before the third decade. A hyperactive vestibuloocular reflex was found in 3 of 11 cases: 2 persons were especially susceptible to motion sickness. Persons with this KCNQ4 mutation showed congenital, progressive high-frequency impairment without substantial loss of speech recognition during the first decades of life. PMID- 11915883 TI - Simulation of organismic morphology and behavior by synthetic poly-alpha-amino acids. AB - Experiments imitating spontaneous geothermal occurrences have yielded most of the amino acids found in protein. All of the amino acids found in protein are simultaneously condensed, by heating in a range of appropriate conditions, to polymers which have many of the properties of proteins. These properties include molecular weights of many thousand, digestibility by proteolytic enzymes, and catalytic activities. One of the other properties is the tendency to form structured units; these units have many of the attributes of biocells. The processes indicated, and others, comprise a conceptual continuum which, according to accumulated information, must have occurred under the conditions existing in regions of the primitive Earth. PMID- 11915885 TI - Microbial contaminants in the interiors of spacecraft components. AB - The hazard from microbiological contamination in the interiors of components of planetary-impacting spacecraft has been generally recognized, but techniques for experimentally evaluating the significance of this hazard have been unavailable. A goal in sterilizing early Mars-impacting payloads is the elimination of the possibility that a microbial stowaway will cause the extraterrestrial-life detection techniques falsely to indicate existence of Martian life, particularly if none exists. Another goal is the prevention of changes in Martian ecology. Several promising life-detection techniques were evaluated in an experimental study of methods to detect microbial populations inside spacecraft-type materials after pulverization or being dissolved. The type of material inspected has a greater effect on the sensitivity of the detection method than any other parameter studied. Through culturing abraded particles, a bacillus inoculum of 100 spores per milliliter can be detected in solid propellants. Fifty percent of an inoculum can be recovered from some solids soluble in nontoxic solvents. For many of the solids studied, however, the culturing techniques used were unable to detect inoculums of 10(6) spores per milliliter of solid. The techniques other than culturing did not discriminate between living and dead cells. As with products of the food and pharmaceutical industries, assurance of sterility in the final product never rests in a negative sterility test on a copy of the product. A negative sterility test is certainly most meaningful for a product for which a very sensitive sterility test has been developed. When a sensitive final-product sterility test is not available, assurance of sterility depends more heavily on other information such as the manufacturing history of the product. PMID- 11915884 TI - The possibility of life in outer space. AB - Experiments were carried out dealing with the effect of extreme factors on microorganisms. Methods for microbiological analysis of meteorites were developed. The effect of temperature, from -20 degrees C to +15 degrees C, on trypsine activity was studied. It was shown that at about -3 degrees C a definite intramolecular reorganization occurs in the enzyme, leading to decreased activation energy. Previously reported resistance of microorganisms to high vacuum was confirmed. New species were tested at l0(-8)-10(-9) mm Hg. It was shown that very thin metal and metal oxide films, as well as films from other substances, may fully protect microorganisms from the hazardous effect of UV rays. By using an artificial climate, camera forms of microorganisms were selected which have the highest resistance to the whole complex of Martian climatic factors. Pigmented and spore-forming microorganisms resist UV up to a dose of 4.04 x 10(8) erg/cm2. There are microorganisms on Earth which show some activity in soil at a relative humidity of 0.096-0.196%. Preliminary data permit one to assume the possibility of existence of Earth-like forms on Mars. The majority of meteorites are accessible to Earth microorganisms. While choosing a meteorite for microbiological analysis it is necessary to consider the soil and climatic factors of the country where the latter has been found. PMID- 11915886 TI - Survival and growth of potential microbial contaminants in severe environments. AB - Studies conducted in our laboratory have established that a number of potential soil microbes could survive a simulated Martian environment. In view of the uncertainty of the Martian environment and the importance of noncontamination of extraterrestrial bodies, studies were performed with common soil microbes to determine the minimal environment necessary to produce a complete growth cycle. The effects of diurnal temperature cycling (+25 degrees C to -65 degrees C) and of limiting concentrations of moisture and oxygen on spore germination, vegetative growth and sporulation of Bacillus cereus and B. subtilis have been determined. The results indicated that diurnally temperature-cycled heat-shocked spores of B. cereus in the simulated Martian atmosphere: 1) survived when the moisture concentration was < or = 4%; 2) germinated but became nonviable when the moisture concentration was > 4 < 8%; 3) germinated with subsequent vegetative growth when a) the moisture concentration was > or = 8%, and b) the concentration was > or = 6% and the partial pressure of oxygen was 15 mm; 4) sporulated when the moisture concentration was > or = 6% and the partial pressure of oxygen was 15 mm. Similar data for heat-shocked spores of B. subtilis in the simulated Martian atmosphere, diurnally temperature-cycled, indicated that the spores: 1) survived when the moisture concentration was > or = 6%; 2) germinated but became nonviable when the moisture concentration was > 1 < 4%; 3) germinated with subsequent vegetative growth when the moisture concentration was > or = 8%; 4) sporulated when the moisture concentration was > or = 6% and the partial pressure of oxygen was > or = 10 mm. Preliminary studies with B. cereus and B. subtilis spores produced in the simulated Martian environment and reintroduced into this environment indicated that vegetative cell growth and sporulation were normal. These data are discussed with regard to the probability of contamination of extraterrestrial bodies. PMID- 11915887 TI - On the thresholds of gravitational force perception by plants. AB - Changes in growth and development of plants in "weightless" environments is a focus of several biosatellite projects. Implicit in experimental designs is the consideration that the force vectors from vibration and vehicular spin are near or below the thresholds of perception. Thresholds of 10(-3) x g have been assigned based upon results from centrifugal stimulation of organs essentially fixed with respect to earth gravity. A value of 2 x 10(-5) x g was derived from vibrational (discontinuous) acceleration of a clinostat wire on which seedlings were mounted. We have developed apparatus that will enable the imposition upon seedlings of a continuous centrifugal force of constant sign concomitant with the nullification of the directional component of earth gravity. Recent experiments evaluating the responses of Avena roots and shoots in such apparatus indicate a threshold on the order of 10(-3)-10(-2) x g. These values are supported by results of experiments where organs were grown on clinostats whose rotational axes were at various tilt angles with respect to earth gravity; this technique enables minute increments of longitudinal gravitational stimulation as a vector function of the tilt angle, concomitant with compensation for earth gravity. We will discuss the possibility that assigned thresholds of micro-g magnitude are derived as a consequence of undefined forces that are artifacts of apparatus function. PMID- 11915888 TI - A review of evidence for biological material in meteorites. AB - The first carbonaceous chondrite fell in France in 1806, and in the years following analyses for organic matter were made which showed this material to be similar to material of biological origin. But the analyses were not conclusive. Within the last few years, additional work has been done which has proved to be very interesting and somewhat controversial. The composition of the inorganic fraction of these objects is very similar to that of other meteorites, showing that no sorting by sedimentary processes has occurred. For this reason, students of meteorites have believed that the material could not be biological. However, the soluble constituents are similar to those to be expected in a primitive ocean of the earth. Some of the assumed fossils have been shown to be contaminants but others appear to be indigenous. After mineral matters are removed from them they still show evidence of containing carbonaceous material. Micro-paleontologists have also reported material which they believe to be similar to terrestrial fossils. Fatty acids, porphyrins, and nucleic acid bases have been reported and a small optical activity as well. The immense difficulty that students of meteorites have in understanding this situation may be alleviated if it should be proved that some of our stone meteorites, including the carbonaceous ones, are coming from the moon. There seems to be some possibility that this could be true. PMID- 11915889 TI - Paraffinic hydrocarbons in the Orgueil, Murray, Mokoia and other meteorites. AB - Small samples of the Orgueil (Wiik Type I), Murray (Wiik Type II), Mokoia (Wiik Type III) and other meteorites (Alais, Ivuna, Al Rais, Bells, Cold Bokkeveld, Mighei, Nogoya, Santa Cruz, Grosnaja, Kaba, Lance, Ornans, Chainpur, Karoonda, Warrenton, and St. Caprais) were extracted with organic solvents, fractionated by silica gel chromatography, and analyzed by gas chromatography and by a new gas chromatographic-mass spectrometric technique. Only the high molecular weight hydrocarbons eluted by n-heptane were analyzed. The following observations were made: All meteorites contain paraffinic hydrocarbons in varying amounts and with a certain distribution. Normal and isomeric alkanes from Cl5 to about C25 with a maximum at C19-C21 were found in the Orgueil, Murray and Mokoia meteorites. Pristane and phytane, two saturated polyisoprenoid hydrocarbons, known to be degradation products of chlorophyll, were found in all meteorites except the Ornans, although in several (Type I and Type III, except Mokoia) the amounts were very small. No predominance of odd over even carbon-number hydrocarbons was observed except in the C22-C27 normal alkane range of some samples of the Orgueil. The concentration of alkanes in certain samples of the Orgueil was found to be about one order of magnitude lower than in others indicating a significant heterogeneity in the distribution of hydrocarbons. Practically all the meteorites of Type II and some of Type III (Mokoia) gave an essentially identical or very similar hydrocarbon chromatographic pattern, indicating a common origin for the hydrocarbons. The distribution of paraffinic hydrocarbons, including pristane and phytane, in some of these meteorites was found similar to that of microfossil bearing pre-Cambrian rocks 2 x 10(9) years old or older. The Orguiel was similar to the Gunflint chert and Type II meteorites were similar to the Soudan rock. It remains to be seen whether these hydrocarbons were formed on the earth-moon system as suggested by Urey, on the asteroids, in the solar nebula, or whether they may be the result of a unique terrestrial contamination process. PMID- 11915890 TI - Hormone replacement therapy and cardiovascular disease: a statement for healthcare professionals from the American Heart Association. PMID- 11915905 TI - Welfare reform and substance abuse: innovative state strategies. AB - This issue brief highlights key facts about the impact of substance abuse on welfare reform and recipients of Temporary Assistance to Needy Families, or TANF. After outlining some of the data on the incidence of substance abuse as well as its costs and treatment, it concludes by describing innovative state welfare programs attempting to lower barriers to employment and self-sufficiency. PMID- 11915907 TI - Exercise and aging. Proceedings of the 11th International Conference on the Biochemistry of Exercise. Little Rock, Arkansas, USA, June 2000. PMID- 11915906 TI - Pharmacy benefits: new concepts in plan design. AB - This issue brief examines changes to prescription drug benefit coverage in large employer plans and implications for a Medicare prescription drug benefit. The brief discusses reasons behind employer benefit plan redesign and recent coverage trends, as well as potential paths to modernize benefits. Information is based on a literature review and conversations and interviews with employers, benefit consultants, and pharmacy benefit management executives. PMID- 11915908 TI - Pathophysiology of body composition and metabolic abnormalities in HIV-infection: therapeutic implications. AB - Changes in body composition and metabolism have been a central feature of HIV infection from the outset--initially, as the wasting syndrome and, more recently, as metabolic and body fat redistribution syndromes associated with antiretroviral (ARV) therapy. Here, advances in physiologic and biochemical understanding of these conditions are reviewed. First, the pathophysiology of wasting in HIV-1 infection is discussed, focusing on the failure of nutrients to increase lean tissue ("anabolic block") and the role of hypogonadism. Results of anabolic interventions, including recombinant growth hormone, testosterone, and progressive resistance exercise, are presented. Next, ARV-associated disorders are reviewed, including lipoatrophy and an hypothesized "mitochondrial toxicity." The possibility of establishing pathogenesis in vivo in humans, by direct measurement of mitochondrial DNA synthesis and adipocyte proliferation, is discussed. In summary, important advances have occurred toward the goal of explaining body composition and metabolic abnormalities associated with HIV disease. PMID- 11915909 TI - Effects of resistance training on the rate of muscle protein synthesis in frail elderly people. AB - Advancing age is associated with a reduction in skeletal muscle protein, muscle strength, muscle quality, and chemical modifications that may impair protein function. Sarcopenia has been coupled with physical disability, frailty, and a loss of independent function. Using stable isotope tracer methodologies and mass spectrometric detection, we observed: (a) 76-92-year-old physically frail and 62 74-year-old middle-age adults have lower mixed muscle protein synthetic rates than 20-32-year-old men and women; (b) 2 weeks and 3 months of weightlifting exercise increased the synthetic rate of myosin heavy chain (MHC) and mixed muscle proteins to a similar magnitude in frail, middle-age, and young women and men; (c) Serum myostatin-immunoreactive protein levels were elevated in physically frail women and were inversely correlated with lean mass. This suggests that the protein synthetic machinery adapts rapidly to increased contractile activity and that the adaptive response(s) are maintained even in frail elders. PMID- 11915910 TI - Muscle protein metabolism and the sarcopenia of aging. AB - Loss of muscle mass, strength, and oxidative capacity accompanies normal aging in humans. The mechanisms responsible for these changes remain to be clearly defined. Muscle protein mass and function depend on protein turnover. Synthesis rate of the major muscle contractile protein, myosin heavy chain (MHC), and transcript levels of fast MHC isoforms decrease in association with strength reductions, while mitochondrial protein synthesis rate declines in parallel with activities of mitochondrial enzymes and maximal oxidative capacity (VO2max). Resistance exercise training increases the synthesis rate of MHC and transcript levels of the slow MHC isoform in older humans, along with increasing muscle strength. The relationship between the synthesis of muscle proteins, and muscle size and function, with aging and exercise training are discussed in this review. PMID- 11915911 TI - Cellular and molecular mechanisms for the bone response to mechanical loading. AB - To define the cellular and molecular mechanisms for the osteogenic response of bone to increased loading, several key steps must be defined: sensing of the mechanical signal by cells in bone, transduction of the mechanical signal to a biochemical one, and transmission of that biochemical signal to effector cells. Osteocytes are likely to serve as sensors of loading, probably via interstitial fluid flow produced during loading. Evidence is presented for the role of integrins, the cell's actin cytoskeleton, G proteins, and various intracellular signaling pathways in transducing that mechanical signal to a biochemical one. Nitric oxide, prostaglandins, and insulin-like growth factors all play important roles in these pathways. There is growing evidence for modulation of these mechanotransduction steps by endocrine factors, particularly parathyroid hormone and estrogen. The efficiency of this process is also impaired in the aged animal, yet what remains undefined is at what step mechanotransduction is affected. PMID- 11915912 TI - Aging and the osteogenic response to mechanical loading. AB - The osteogenic response to mechanical stress is blunted with aging. It has been postulated that this decline in responsiveness is related to (a) a limited ability to engender the strain necessary to reach the bone modeling threshold, due to decreased muscle mass and strength, and/or (b) a decline in certain hormones or growth factors that may interact with mechanical signals to change the sensitivity of bone cells to strain. There is reason to believe that both of these factors contribute to the reduced ability to increase bone mass through exercise with advancing age. Weight-bearing endurance exercise and resistance exercise have both been found to increase bone mass in older women and men. However, exercise training studies involving older individuals have generally resulted in increased bone mineral density only when the exercise is quite vigorous. There is also evidence that the osteogenic response to mechanical loading is enhanced by estrogens. Whether age-related changes in other factors (e.g., other hormones, growth factors, cytokines) also contribute to the reduced responsiveness of the aged skeleton to mechanical loading remains to be investigated. PMID- 11915913 TI - Translational control of protein synthesis: implications for understanding changes in skeletal muscle mass. AB - Gain or loss of skeletal muscle mass is due largely to the establishment of an imbalance between rates of protein synthesis and degradation. A key determinant of the rate of protein synthesis is translation initiation, a process regulated in part through binding of initiator methionyl-tRNA (met-tRNAi) and messenger RNA (mRNA) to a 40S ribosomal subunit. Either the met-tRNAi or mRNA binding step can become limiting for protein synthesis. Furthermore, the mRNA binding step can modulate translation of specific mRNAs with or without changes in the overall rate of protein synthesis. This report highlights molecular mechanisms involved in mediating control of the mRNA binding step in translation initiation. Particular attention is given to the effect of exercise on this step and to how the branched-chain amino acid leucine stimulates muscle protein synthesis after exercise. Potential mechanisms for exercise-induced increase in muscle mass are discussed. PMID- 11915914 TI - Protein metabolism and age: influence of insulin and resistance exercise. AB - Skeletal muscle proteins are constantly being synthesized and degraded, and the net balance between synthesis and degradation determines the resultant muscle mass. Biochemical pathways that control protein synthesis are complex, and the following must be considered: gene transcription, mRNA splicing, and transport to the cytoplasm; specific amino acyl-tRNA, messenger (mRNA), ribosomal (rRNA) availability; amino acid availability within the cell; the hormonal milieu; rates of mRNA translation; packaging in vesicles for some types of proteins; and post translational processing such as glycation and phosphorylation/dephosphorylation. Each of these processes is responsive to the need for greater or lesser production of new proteins, and many states such as sepsis, uncontrolled diabetes, prolonged bed-rest, aging, chronic alcohol treatment, and starvation cause marked reductions in rates of skeletal muscle protein synthesis. In contrast, acute and chronic resistance exercise cause elevations in rates of muscle protein synthesis above rates found in non-diseased rested organisms, which are normally fed. Resistance exercise may be unique in this capacity. This chapter focuses on studies that have used exercise to elucidate mechanisms that explain elevations in rates of protein synthesis. Very few studies have investigated the effects of aging on these mechanisms; however, the literature that is available is reviewed. PMID- 11915915 TI - Contractile properties of aging skeletal muscle. PMID- 11915916 TI - Control of muscle protein breakdown: effects of activity and nutritional states. AB - We propose that there is a link between muscle protein synthesis and breakdown that is regulated, in part, through maintenance of the free intracellular pool of essential amino acids. For example, we propose that muscle protein breakdown is paradoxically elevated in the anabolic state following resistance exercise in part because the even greater stimulation of synthesis would otherwise deplete this pool. Thus, factors regulating muscle protein breakdown must be evaluated in the context of the prevailing rate of muscle protein synthesis. Further, the direct effect of factors on breakdown may depend on the physiological state. For example, local hyperinsulinemia suppresses accelerated muscle protein breakdown after exercise, but not normal resting breakdown. Thus, factors regulating muscle protein breakdown in human subjects are complex and interactive. PMID- 11915917 TI - Control of muscle protein synthesis as a result of contractile activity and amino acid availability: implications for protein requirements. AB - The major anabolic influences on muscle are feeding and contractile activity. As a result of feeding, anabolism occurs chiefly by increases in protein synthesis with minor changes in protein breakdown. Insulin has a permissive role in increasing synthesis, but the availability of amino acids is crucial for net anabolism. We have investigated the role of amino acids in stimulating muscle protein synthesis, the synergy between exercise and amino acid availability, and some of the signaling elements involved. The results suggest that muscle is acutely sensitive to amino acids, that exercise probably increases the anabolic effects of amino acids by a separate pathway, and that for this reason it is unlikely that accustomed physical exercise increases protein requirements. PMID- 11915918 TI - mTOR-dependent control of skeletal muscle protein synthesis. AB - Muscle mass is influenced by many factors including genetically programmed changes, hormonal state, level of activity, and disease processes. Ultimately, whether or not a muscle hypertrophies or atrophies is determined by a simple relationship between the rates of protein synthesis and degradation. When synthesis exceeds degradation, the muscle hypertrophies, and vice versa. In contrast to this simple relationship, the processes that control muscle protein synthesis and degradation are complex. Recently, significant progress has been made in understanding the biochemical mechanisms that control the rate of translation initiation, which is generally the limiting phase in protein synthesis. PMID- 11915919 TI - Cellular adaptations to endurance exercise: master athletes. PMID- 11915920 TI - The aging cardiovascular system: changes in autonomic function at rest and in response to exercise. AB - Tonic vagal modulation of cardiac period (R-R interval) decreases with advancing age, but is greater in middle-aged and older adults who habitually perform aerobic exercise compared with their sedentary peers. Cardiovagal baroreflex sensitivity also declines markedly with age in sedentary adults but only 50% as much in regularly exercising adults. In previously sedentary middle-aged and older adults, a 3-month program of moderate aerobic exercise increases cardiovagal baroreflex sensitivity by 25%. Tonic (basal) sympathetic nervous system (SNS) activity increases with advancing age in both sedentary and habitually exercising adults. Despite this, SNS beta-adrenergic support of energy metabolism (resting metabolic rate--RMR) declines with age in sedentary individuals. However, SNS beta-adrenergic support of RMR is maintained with age in endurance exercise-trained adults and therefore is much greater in middle-aged and older individuals who exercise regularly compared with their sedentary peers. Thus, regular aerobic (endurance) exercise modulates selective age-associated impairments in autonomic nervous system-physiological function. PMID- 11915921 TI - Master athletes. AB - Over the past 3 decades, there has been a continued increase in the number of "older" participants in sporting events such as running, swimming, cycling, rowing, and weightlifting. Some master athletes come from a background with years of training and competition experience, while others have only begun to compete as they approach middle-aged and older. The majority of what we currently know about master athletes and aging has been gained from both cross-sectional and longitudinal testing and re-testing master athletes and recreational athletes. The focus of this paper is on the physiological profile of athletes and individuals performing regular exercise training. Physiological data from elite and non-elite, recreational, sedentary, and senior athletes clearly indicate that human skeletal muscle has a high degree of plasticity that is maintained late into life. Muscle fiber protein expression and single muscle fiber contractile properties are greatly influenced by exercise training. It appears that skeletal muscle can quickly adapt to accommodate a wide range of functionality to meet the demands (or lack of demands) placed upon it. PMID- 11915922 TI - Energy expenditure and aging: effects of physical activity. AB - Aging is associated with a decline in daily energy expenditure that is disproportionately greater than the decline in daily energy intake. Collectively, these events can create a "positive" energy balance, secondary gains in central and total body fat, and a subsequently higher risk of morbidity and mortality. Participation in regular physical activity is a logical strategy to attenuate the decline in energy expenditure with aging, as physical activity can comprise between 10-50% of an older person's daily energy expenditure. Understanding the influence of regular physical activity on energy expenditure with advancing age is clinically relevant, particularly since estimates predict that nearly 25% of the population will be > or = 65 years of age by the year 2030. This brief review will focus on the current state of aging, energy expenditure, and physical activity literature. Topics to be addressed include: (a) measurement of physical activity in older adults; (b) aging and physical inactivity; and (c) influence of regular aerobic exercise on resting metabolic rate (RMR), thermic effect of food (TEF), and non-exercising physical activity. PMID- 11915923 TI - Effects of activity on growth factor expression. AB - For some time, it has been appreciated that muscle mass is regulated locally as well as systemically. We have cloned the cDNA of two isoforms of IGF-1, which are derived from the IGF-1 gene by alternate splicing. The expression of one of these was only detectable after mechanical stimulation. For this reason, this has been called mechano growth factor (MGF). The MGF is not glycosylated, is smaller, and has a shorter half-life in the unbound state than the systemic liver type IGF-1. As the result of a reading frame shift the MGF peptide also has a different C terminal sequence and thus has different binding protein/receptor affinities. Another splice variant (muscle L.IGF-I) is expressed in muscle during rest but is also upregulated by exercise. The latter is similar to the systemic liver type IGF-1. The evidence suggests that MGF has a high potency for inducing local protein synthesis and preventing apoptosis and therefore has an important role in local tissue repair and remodeling. Our physiological experiments show that stretch and particularly stretch combined with electrical stimulation, rather than stimulation alone are important in inducing MGF expression. The mechanotransduction mechanism is believed to involve the muscle cytoskeleton. During aging, the production of growth hormone and IGF-1 by the liver declines markedly. The discovery of MGF and muscle IGF-1 provides a link between physical activity and gene expression. This underlines the need for the elderly to remain active as the locally produced growth factors supplement the circulating IGF-1 levels. PMID- 11915924 TI - Physiology of aging. AB - Although physiological deterioration occurs with advancing adult age, the interpretation of cross-sectional and longitudinal studies of age-changes in physiological processes are often complicated by confounders unrelated to aging. Age-associated disease is a major cause of physiological deterioration. To avoid this factor, only subjects free of discernible disease are used in many studies of what is referred to as "normal" aging. However, it is important to recognize that age-associated disease is not only a common occurrence but also an integral part of aging. Increasingly, gene-environment interactions are recognized as playing a major role in age-associated physiological deterioration, and there is great individual variation in both the rate of aging and the occurrence of age associated physiological deterioration. PMID- 11915925 TI - Effects of aging on regulation of muscle contraction at the motor unit, muscle cell, and molecular levels. AB - Rodent motor units, muscle fibers, and motor proteins undergo significant aging related changes. Such changes include spatial organization and physiological properties of fast- and slow-twitch single motor units, regulation of contractile speed and force generation capacity at the muscle fiber level, and altered functional properties of the motor protein myosin. In addition to specific changes, there also appears to be a "disorganization" of the coordinated expression of contractile, sarcoplasmic reticular, and mitochondrial protein isoforms in aging skeletal muscle. This is suggested to have a strong impact on aging-related impairments in muscle function in addition to the changes in specific muscle proteins. PMID- 11915926 TI - Adaptive potentials of skeletal muscle in young and aging rats. AB - We compared responses of the fast extensor digitorum longus (EDL) and tibialis anterior (TA) muscles in young (15-week) and aging (101-week) male Brown Norwegian rats to 50 days of chronic low-frequency stimulation (CLFS, 10 Hz, 10 hours/day). After 50 days of CLFS, the EDL muscles of the young (22-week) and aging (108-week) rats displayed similar increases in type IIA fibers, relative concentration of myosin heavy chain MHCIIa, elevations in mitochondrial citrate synthase and 3-hydroxyacyl-CoA dehydrogenase activities, and similar decreases in glycolytic enzyme activities (glyceraldehydephosphate dehydrogenase, lactate dehydrogenase). TA muscle in young rats contained a few cytochrome c oxidase negative (COX-) type I fibers. Their number was approximately 2-fold elevated by CLFS. Conversely, aging muscle, which contained a slightly higher amount of COX- fibers than young TA muscle, responded to CLFS with a significant decrease in COX fibers. The appearance of small COX-positive type I fibers in stimulated aging muscle indicated that regenerating type I fibers "diluted" the COX-deficient fiber population. PMID- 11915927 TI - The molecular responses of skeletal muscle satellite cells to continuous expression of IGF-1: implications for the rescue of induced muscular atrophy in aged rats. AB - Approximately 50% of humans older than 85 years have physical frailty due to weak skeletal muscles. This indicates a need for determining mechanisms to combat this problem. A critical cellular factor for postnatal muscle growth is a population of myogenic precursor cells called satellite cells. Given the complex process of sarcopenia, it has been postulated that, at some point in this process, a limited satellite cell proliferation potential could become rate-limiting to the regrowth of old muscles. It is conceivable that if satellite cell proliferative capacity can be maintained or enhanced with advanced age, sarcopenia could potentially be delayed or prevented. Therefore, the purposes of this paper are to describe whether IGF-I can prevent muscular atrophy induced by repeated cycles of hindlimb immobilization, increase the in vitro proliferation in satellite cells from these muscles and, if so, the molecular mechanisms by which IGF-I mediates this increased proliferation. Our results provide evidence that IGF-I can enhance aged muscle regrowth possibly through increased satellite cell proliferation. The results also suggest that IGF-I enhances satellite cell proliferation by decreasing the cell cycle inhibitor, p27Kip1, through the PI3'-K/Akt pathway. These data provide molecular evidence for IGF-I's rescue effect upon aging associated skeletal muscle atrophy. PMID- 11915928 TI - Influence of aging and exercise on endocrine function. PMID- 11915929 TI - Overview of glucose metabolism and aging. PMID- 11915930 TI - Mechanisms of muscle insulin resistance in obese individuals. AB - We previously reported that insulin resistance in skeletal muscle of obese individuals was associated with decreases in insulin signal transduction and tyrosine kinase activity of the insulin receptor. Herein is reviewed the recently published data supporting the hypothesis that protein kinase C (PKC) phosphorylates the insulin receptor on serine/threonine residues to decrease tyrosine kinase activity and cause insulin resistance. Treatment of insulin receptors from obese subjects with alkaline phosphatase restored tyrosine kinase activity, suggesting that the reduced activity was a result of hyperphosphorylation of the receptor. Incubating human muscle fiber strips with PKC inhibitors restored insulin action in muscle of obese patients, while activating PKC with a phorbol ester caused insulin resistance in muscle from lean control patients. The beta isoform of PKC was elevated in muscle of obese, insulin-resistant patients. These data are consistent with the hypothesis that elevated PKC activity may cause insulin resistance by phosphorylating the insulin receptor to decrease tyrosine kinase activity. PMID- 11915931 TI - Regulation of muscle glucose transport during exercise. AB - In the present short review some factors affecting glucose utilization during exercise in skeletal muscle will be briefly described. Special focus will be put on the glucose transport step across the sarcolemma. Glucose transporters (GLUT4) are expressed at a surprisingly similar level in the different muscle fiber types in human skeletal muscle in contrast to findings in the rat. When working at the same absolute work load muscle glucose transport is decreased in trained compared with untrained muscle in part due to a decrease in GLUT4 translocation to the sarcolemma in trained muscle. However, when trained and untrained muscle are stressed severely by a workload taxing 100% of their peak oxygen uptake in a glycogen-depleted state, then glucose uptake is larger in trained than in untrained muscle and correlates with muscle GLUT4 content. Finally, the possible role of the AMP-activated protein kinase (AMPK) in regulating glucose uptake during exercise is discussed. It is indicated that at present no experiments definitively link activation of AMPK to activation of muscle glucose transport during exercise. PMID- 11915932 TI - Insulin action, training, and aging. PMID- 11915933 TI - Effect of aging on glucose and lipid metabolism during endurance exercise. AB - Endurance exercise increases the use of endogenous fuels to provide energy for working muscles. Elderly subjects oxidize more glucose and less fat during moderate intensity exercise. This shift in substrate use is presumably caused by age-related changes in skeletal muscle, including decreased skeletal muscle respiratory capacity, because adipose tissue lipolysis and plasma fatty acid availability are not rate limiting. Endurance training in elderly subjects increases muscle respiratory capacity, decreases glucose production and oxidation, and increases fat oxidation thereby correcting or compensating for the alterations in substrate oxidation associated with aging. PMID- 11915934 TI - Molecular characteristics of aged muscle reflect an altered ability to respond to exercise. AB - Studies have been performed in humans to identify changes in gene expression that may account for the relatively weak and variable response of aged muscle to resistance exercise. The gene expression profile of skeletal muscle from elderly (62-75 years old) compared to younger (20-30 years old) men demonstrated elevated expression of genes typical of a stress or damage response. The expression of the majority of these genes was unaffected by a single bout of high-intensity resistance exercise in elderly subjects but was altered acutely by exercise in younger subjects so as to approach the pre-exercise levels observed in older subjects. The inability of muscle from elderly subjects to respond to resistance exercise was also apparent in the expression of inflammatory response genes, which increased within 24 hours of the exercise bout only in younger subjects. Othergenes with potentially important roles in the adaptation of muscle to exercise, showed a similar or even more robust response in older compared to younger subjects. Taken together, these results may help to explain the variable hypertrophic response of muscle from older individuals to resistance training. PMID- 11915935 TI - Skeletal muscle fatty acid transport and transporters. AB - While it has long been assumed that long chain fatty acids (LCFA) can freely diffuse across the plasma membrane, recent work has shown that LCFA uptake also involves a protein-mediated mechanism. Three putative LCFA transporters have been identified (FABPpm, FATP, and FAT/CD36), and all are expressed in rodent and human muscles. In a new model system (giant vesicles), we have demonstrated that (a) LCFA transport rates are scaled with the oxidative capacity of heart and muscle, (b) only FABPpm and FAT/CD36, but not FATP1, correlate with vesicular LCFA transport, and (c) LCFA transport can be increased by increasing (1) the FAT/CD36 protein of muscle (chronic adaptation) or (2) via the translocation of FAT/CD36 from an intracellular pool to the plasma membrane during muscle contraction (acute adaptation). PMID- 11915936 TI - Plasma triglyceride metabolism in humans and rats during aging and physical inactivity. AB - Physical activity often declines with age because of a reduction in the spontaneous activities of daily living and because of less intense exercise. In controlled studies of young rats, it was shown that physical activities associated with walking and standing were especially important for maintaining a high level of lipoprotein lipase (LPL) activity in postural skeletal muscles (slow-twitch oxidative muscles). More intense contractions during run training were important for a high LPL activity in the fast-twitch glycolytic muscles. Aging also causes a fiber type-specific decrease of skeletal muscle LPL activity and LPL protein in weight-bearing skeletal muscles (and no aging effect in glycolytic muscles). Thus, contractile inactivity may be a significant factor causing sub-optimal triglyceride metabolism in skeletal muscles during both unloading in young animals and aging. Measurements of plasma LPL activity, plasma triglyceride (TG) clearance rates, postprandial hypertriglyceridemia after oral fat tolerance tests, and fasting TG levels were generally indicative of reduced plasma TG metabolism during middle or old age. In contrast, older endurance trained individuals had a favorable blood lipid profile compared to age-matched or young controls, even when the controls were not overweight. Therefore, the poor TG metabolism that is frequently associated with aging may be caused by some of the same processes that lower skeletal muscle LPL activity of young sedentary individuals. PMID- 11915937 TI - Factor Xa--a promising target for drug development. PMID- 11915938 TI - Sterol carrier protein-2: structure reveals function. AB - The multiple actions of sterol carrier protein-2 (SCP-2) in intracellular lipid circulation and metabolism originate from its gene and protein structure. The SCP x/pro-SCP-2 gene is a fusion gene with separate initiation sites coding for 15 kDa pro-SCP-2 (no enzyme activity) and 58-kDa SCP-x (a 3-ketoacyl CoA thiolase). Both proteins share identical cDNA and amino acid sequences for 13-kDa SCP-2 at their C-termini. Cellular 13-kDa SCP-2 derives from complete, posttranslational cleavage of the 15-kDa pro-SCP-2 and from partial posttranslational cleavage of 58-kDa SCP-x. Putative physiological functions of SCP-2 have been proposed on the basis of enhancement of intermembrane lipid transfer (e.g., cholesterol, phospholipid) and activation of enzymes involved in fatty acyl CoA transacylation (cholesterol esters, phosphatidic acid) in vitro, in transfected cells, and in genetically manipulated animals. At least four important SCP-2 structural domains have been identified and related to specific functions. First, the 46-kDa N terminal presequence present in 58-kDa SCP-x is a 3-ketoacyl-CoA thiolase specific for branched-chain acyl CoAs. Second, the N-terminal 20 amino acid presequence in 15-kDa pro-SCP-2 dramatically modulates the secondary and tertiary structure of SCP-2 as well as potentiating its intracellular targeting coded by the C-terminal peroxisomal targeting sequence. Third, the N-terminal 32 amino acids form an amphipathic a-helical region, one face of which represents a membrane-binding domain. Positively charged amino acid residues in one face of the amphipathic helices allow SCP-2 to bind to membrane surfaces containing anionic phospholipids. Fourth, the hydrophobic faces of the N-terminal amphipathic a helices along with beta strands 4, 5, and helix D form a ligand binding cavity able to accommodate multiple types of lipids (e. g., fatty acids, fatty acyl CoAs, cholesterol, phospholipids, isoprenoids). Two-dimensional 1H-15N heteronuclear single quantum coherence spectra of both apo-SCP-2 and of the 1:1 oleate-SCP-2 complex, obtained at pH 6.7, demonstrated the homogenous formation of holo-SCP-2. While comparison of the apo- and holoprotein amide fingerprints revealed about 60% of the resonances remaining essentially unchanged, 12 assigned amide residues underwent significant chemical-shift changes upon oleic acid binding. These residues were localized in three regions: the juncture of helices A and B, the mid-section of the beta sheet, and the interface formed by the region of beta strands 4, 5, and helix D. Circular dichroism also showed that these chemical-shift changes, upon oleic acid binding, did not alter the secondary structure of SCP-2. The nuclear magnetic resonance chemical shift difference data, along with mapping of the nearby hydrophobic residues, showed the oleic acid-binding site to be comprised of a pocket created by the face of the beta sheet, helices A and B on one end, and residues associated with beta strands 4, 5, and helix D at the other end of the binding cavity. Furthermore, the hydrophobic nature of the previously ill-defined C-terminus suggested that these 20 amino acids may form a 'hydrophobic cap' which closes around the oleic acid upon binding. Thus, understanding the structural domains of the SCP-x/pro SCP-2 gene and its respective posttranslationally processed proteins has provided new insights into their functions in intracellular targeting and metabolism of lipids. PMID- 11915939 TI - Evidence of undiscovered cell regulatory mechanisms: phosphoproteins and protein kinases in mitochondria. AB - The finding that mitochondria contain substrates for protein kinases lead to the discovery that protein kinases are located in the mitochondria of certain tissues and species. These include pyruvate dyhydrogenase kinase, branched-chain alpha ketoacid dehydrogenase kinase, protein kinase A, protein kinase Cdelta, stress activated kinase and A-Raf as well as unidentified kinases. Recent evidence suggests that mitochondrial protein kinases may be involved in physiological processes such as apoptosis and steroidogenesis. Additionally, the novel finding of low-molecular-weight GTP-binding proteins in mitochondria suggests the possibility that these may interact with mitochondrial protein kinases to regulate the activity of mitochondrial effector proteins. The fact that there are components of cellular regulatory systems in mitochondria indicates the exciting possibility of undiscovered systems regulating mitochondrial physiology. PMID- 11915940 TI - The murine complement regulator Crry: new insights into the immunobiology of complement regulation. AB - Complement has an important role in inflammation and in the normal function of the immune system. Activated complement fragments have the capacity to bind and damage self-tissues. Cells from vertebrates express on their surface regulators of complement activation that protect them from the deleterious effects of cell bound complement fragments. Abnormalities in these regulators of complement activation may participate in the pathogenesis of autoimmune diseases and inflammatory disorders. Murine Crry is one of these regulators that inhibits the activation of the third component of complement and protects self-tissues from complement-mediated damage. Experimental work on Crry has increased our understanding of the immunobiology of complement regulation and the potential role of complement and complement inhibitors in the development and treatment of human diseases. PMID- 11915942 TI - DNA methylation and the regulation of gene transcription. AB - The regulation of gene transcription is not simply dependent on the presence or absence of DNA-binding transcription factors that turn genes on or off, but also involves processes determining the ability of transcription factors to gain access to and bind their target DNA. Methylation of DNA cytosine bases leads to the inaccessibility of DNA regulatory elements to their transcription factors by a number of mechanisms. Our understanding of DNA methylation has advanced rapidly in recent years with the identification of an increasingly large number of novel proteins involved in this process. These include methylcytosine-binding proteins as well as additional members of the DNA methyltransferase family. The creation of mice with targeted deletions in a number of genes involved in DNA methylation has further elucidated the functions of many of these proteins. The characterization of complexes that contain proteins known to be involved in DNA methylation has led to the identification of additional proteins, especially those involved in histone deacetylation, indicating that DNA methylation and histone deacetylation very likely act in a synergistic fashion to regulate gene transcription. Finally, the implication of DNA methylation in tumorigenesis and the realization that some congenital diseases are caused by deficiency of proteins involved in DNA methylation has confirmed the importance of this process in regulating gene expression. PMID- 11915941 TI - Immunotherapy against antigenic tumors: a game with a lot of players. AB - Our understanding of how immune responses are generated and regulated drives the design of possible immunotherapies for cancer patients. For that reason, we first describe briefly the actual immunological theories and their common perspectives about cancer vaccine development. Second, we describe cancer vaccines that are able to induce tumor-specific immune responses in cancer patients. However, these responses are not always followed by tumor rejection. At the end of the review, we discuss two possible reasons that might explain this dichotomy of cancer immunology. First, the immune response generated, although detectable, may not be quantitatively sufficient to reject the tumor. Second, the tumor microenvironment may modulate tumor cell susceptibility to the systemic immune response induced by the immunization. Finally, we discuss what, in our opinion, might be the best way to improve cancer vaccine strategies and how the relationship between the tumor and its surroundings might be studied in more details. PMID- 11915943 TI - Pathogenesis of Plasmodium falciparum malaria: the roles of parasite adhesion and antigenic variation. AB - Malaria results in up to 2.5 million deaths annually, with young children and pregnant women at greatest risk. The great majority of severe disease is caused by Plasmodium falciparum. A characteristic feature of infection with Pfalciparum is the accumulation or sequestration of parasite-infected red blood cells (RBCs) in various organs, such as the brain, lung and placenta, and together with other factors is important in the pathogenesis of severe forms of malaria. Sequestration results from adhesive interactions between parasite-derived proteins expressed on the surface of infected RBCs and a number of host molecules on the surface of endothelial cells, placental cells and uninfected RBCs. Some receptors for parasite adhesion have been implicated in particular malaria syndromes, such as intercellular adhesion molecule 1 in cerebral malaria and chondroitin sulfate A and hyaluronic acid in placental infection. The principal parasite ligand and antigen on the RBC surface, P falciparum erythrocyte membrane protein 1 encoded by a multigene family termed var, is clonally variant, enabling evasion of specific immune responses. An understanding of these host-parasite interactions in the context of clinical disease and immunity may reveal potential targets to prevent or treat severe forms of malaria. PMID- 11915944 TI - Role of bombesin-related peptides in the mediation or integration of the stress response. AB - In addition to the relatively well established role of corticotropin-releasing hormone (CRH) and arginine-vasopressin (AVP) in the mediation of the stress response, there is reason to believe that bombesin-like peptides (BN-LPs) may also contribute to the mediation or integration of these responses and thus might be considered as putative 'stress peptides'. This review provides evidence supporting this contention by showing that (i) BN-LPs are present at brain sites known to be activated by stressors, (ii) stressor exposure alters utilization of BN-related peptides, (iii) exogenous BN administration mimics the endocrine, autonomic and/or behavioral effects elicited by stressors, and (iv) antagonism of BN action attenuates the behavioral and/or neurochemical effects of stressors or of exogenously administered peptide. The evidence presented also suggests that BN LPs mediate their stress-relevant effects through activation of CRH and/or AVP neurons. Several hypothetical mechanisms for such peptidergic interactions are discussed as to the implications of considering BN-LPs as 'stress peptides'. PMID- 11915945 TI - Pepsinogens, progastricsins, and prochymosins: structure, function, evolution, and development. AB - Five types of zymogens of pepsins, gastric digestive proteinases, are known: pepsinogens A, B, and F, progastricsin, and prochymosin. The amino acid and/or nucleotide sequences of more than 50 pepsinogens other than pepsinogen B have been determined to date. Phylogenetic analyses based on these sequences indicate that progastricsin diverged first followed by prochymosin, and that pepsinogens A and F are most closely related. Tertiary structures, clarified by X-ray crystallography, are commonly bilobal with a large active-site cleft between the lobes. Two aspartates in the center of the cleft, Asp32 and Asp215, function as catalytic residues, and thus pepsinogens are classified as aspartic proteinases. Conversion of pepsinogens to pepsins proceeds autocatalytically at acidic pH by two different pathways, a one-step pathway to release the intact activation segment directly, and a stepwise pathway through a pseudo-pepsin(s). The active site cleft is large enough to accommodate at least seven residues of a substrate, thus forming S4 through S'3 subsites. Hydrophobic and aromatic amino acids are preferred at the P1 and P'1 positions. Interactions at additional subsites are important in some cases, for example with cleavage of kappa-casein by chymosin. Two potent naturally occurring inhibitors are known: pepstatin, a pentapeptide from Streptomyces, and a unique proteinous inhibitor from Ascaris. Pepsinogen genes comprise nine exons and may be multiple, especially for pepsinogen A. The latter and progastricsin predominate in adult animals, while pepsinogen F and prochymosin are the main forms in the fetus/infant. The switching of gene expression from fetal/infant to adult-type pepsinogens during postnatal development is noteworthy, being regulated by several factors, including steroid hormones. PMID- 11915946 TI - Modulation by flavonoids of cell multidrug resistance mediated by P-glycoprotein and related ABC transporters. AB - Cancer cell resistance to chemotherapy is often mediated by overexpression of P glycoprotein, a plasma membrane ABC (ATP-binding cassette) transporter which extrudes cytotoxic drugs at the expense of ATP hydrolysis. P-glycoprotein (ABCB1, according to the human gene nomenclature committee) consists of two homologous halves each containing a transmembrane domain (TMD) involved in drug binding and efflux, and a cytosolic nucleotide-binding domain (NBD) involved in ATP binding and hydrolysis, with an overall (TMD-NBD)2 domain topology. Homologous ABC multidrug transporters, from the same ABCB family, are found in many species such as Plasmodiumfalciparum and Leishmania spp. protozoa, where they induce resistance to antiparasitic drugs. In yeasts, some ABC transporters involved in resistance to fungicides, such as Saccharomyces cerevisiae Pdr5p and Snq2p, display a different (NBD-TMD)2 domain topology and are classified in another family, ABCG. Much effort has been spent to modulate multidrug resistance in the different species by using specific inhibitors, but generally with little success due to additional cellular targets and/or extrusion of the potential inhibitors. This review shows that due to similarities in function and maybe in three dimensional organization of the different transporters, common potential modulators have been found. An in vitro 'rational screening' was performed among the large flavonoid family using a four-step procedure: (i) direct binding to purified recombinant cytosolic NBD and/or full-length transporter, (ii) inhibition of ATP hydrolysis and energy-dependent drug interaction with transporter-enriched membranes, (iii) inhibition of cell transporter activity monitored by flow cytometry and (iv) chemosensitization of cell growth. The results indicate that prenylated flavonoids bind with high affinity, and strongly inhibit drug interaction and nucleotide hydrolysis. As such, they constitute promising potential modulators of multidrug resistance. PMID- 11915948 TI - The prolyl oligopeptidase family. AB - A group of serine peptidases, the prolyl oligopeptidase family, cannot hydrolyze peptides containing more than about 30 residues. This group is unrelated to the classical trypsin and subtilisin families, and includes dipeptidyl peptidase IV, acylaminoacyl peptidase and oligopeptidase B, in addition to the prototype prolyl oligopeptidase. The recent crystal structure determination of prolyl oligopeptidase (80 kDa) has shown that the enzyme contains a peptidase domain with an alpha/beta hydrolase fold, and its catalytic triad is covered by the central tunnel of an unusual seven-bladed beta-propeller. This domain operates as a gating filter, excluding large, structured peptides from the active site. The binding mode of substrates and the catalytic mechanism differ from that of the classical serine peptidases in several features. The members of the family are important targets of drug design. Prolyl oligopeptidase is involved in amnesia, depression and blood pressure control, dipeptidyl peptidase IV in type 2 diabetes and oligopeptidase B in trypanosomiasis. PMID- 11915949 TI - The coagulation of insect hemolymph. AB - In contrast to both vertebrates and non-insect arthropods, little is known about the coagulation of hemolymph (hemostasis) in insects. We discuss the integration of the hemostatic response with other branches of the insect immune system. We also describe the present stage in the characterization of both soluble and cellular factors that contribute to hemostasis in insects. The factors of the well-characterized clotting cascades of vertebrates, primitive chelicerates and crustaceans are used to assess the implications of sequencing the whole Drosophila genome for searching candidate genes involved in hemostasis. Some striking similarities between blood clotting in vertebrates and the reaction of insect cells involved in hemolymph coagulation have implications for a phylogenetic comparison of hemostasis between divergent animal classes. PMID- 11915950 TI - Double-strand breaks and translocations in cancer. AB - The correct repair of double-strand breaks (DSBs) is essential for the genomic integrity of a cell, as inappropriate repair can lead to chromosomal rearrangements such as translocations. In many hematologic cancers and sarcomas, translocations are the etiological factor in tumorigenesis, resulting in either the deregulation of a proto-oncogene or the expression of a fusion protein with transforming properties. Mammalian cells are able to repair DSBs by pathways involving homologous recombination and nonhomologous end-joining. The analysis of translocation breakpoints in a number of cancers and the development of model translocation systems are beginning to shed light on specific DSB repair pathway(s) responsible for the improper repair of broken chromosomes. PMID- 11915951 TI - Assessment of selenium effects in lotic ecosystems. AB - The selenium literature has grown substantially in recent years to encompass new information in a variety of areas. Correspondingly, several different approaches to establishing a new water quality criterion for selenium have been proposed since establishment of the national water quality criterion in 1987. Diverging viewpoints and interpretations of the selenium literature have lead to opposing perspectives on issues such as establishing a national criterion based on a sediment-based model, using hydrologic units to set criteria for stream reaches, and applying lentic-derived effects to lotic environments. This Commentary presents information on the lotic verse lentic controversy. Recently, an article was published that concluded that no adverse effects were occurring in a cutthroat trout population in a coldwater river with elevated selenium concentrations (C. J. Kennedy, L. E. McDonald, R. Loveridge, and M. M. Strosher, 2000, Arch. Environ. Contam. Toxicol. 39, 46-52). This article has added to the controversy rather than provided further insight into selenium toxicology. Information, or rather missing information, in the article has been critically reviewed and problems in the interpretations are discussed. PMID- 11915952 TI - Dioxin body burdens in operation ranch hand veterans: promotion blocking and cancer causation. AB - The cancer incidence data from the Operation Ranch Hand Study are plotted as a function of seven dioxin body burden classifications from zero through modest levels above background. The three significant cancer incidence peaks and three significant cancer incident troughs observed are used to test a model which specifies dioxin activity as both a promoter blocker and a cancer causation agent. There is a significant, 50% reduction in total cancers other than skin at the highest dioxin body burden level compared to background. There is a significant reduction in prostate cancers in Black veterans, but not in White, as a function of increasing dioxin body burden. PMID- 11915953 TI - In vitro development of parthenogenetic eggs: a fast ecotoxicity test with Daphnia magna? AB - In the present study, the authors investigated and defined development stages of Daphnia magna eggs that could be potential endpoints for sublethal toxicity tests with in vitro parthenogenetic egg cultures. Such an "egg test" could be a cost effective alternative to the classic 21-day test with D. magna. Three main stages of embryonic development were considered: release of the egg external membrane, release of the internal membrane, and separation of the caudal spine. The first embryonic stage was attained approximately 30 h after transfer of eggs from ovaries to the brood chamber (considered as time zero), the second stage at 48 h, and the third stage at 68 h. Embryonic development was considered completed with the caudal spine separation. Thereafter, juveniles were able to swim in the water column. Egg mortality, duration of each egg stage, egg diameter, and egg abnormalities were investigated as potential endpoints. In vitro tests were carried out with several toxicants (DBS, 3,4-DCA, cadmium, and copper) and with acid mine drainage, sensitivity generally being higher than with the accepted chronic 21-day test with D. magna. PMID- 11915947 TI - Recent advances in the genetics of schizophrenia. AB - The genetic etiology of schizophrenia, a common and debilitating psychiatric disorder, is supported by a wealth of data. Review of the current findings suggests that considerable progress has been made in recent years, with a number of chromosomal regions consistently implicated by linkage analysis. Three groups have shown linkage to 1q21-22 using similar models, with HLOD scores of 6.5, 3.2, and 2.4. Other replicated loci include 13q32 that has been implicated by two independent groups with significant HLOD scores (4.42) or NPL values (4.18), and 5pl4.1-13.1, 5q21-33, 8p2l-22, and 10p11-15, each of which have been reported as suggestive by at least three separate groups. Different studies have also replicated evidence for a modest number of candidate genes that were not ascertained through linkage. Of these, the greatest support exists for the DRD3 (3q13.3), HTR2A (13q14.2), and CHRNA7 (15q13-q14) genes. The refinement of phenotypes, the use of endophenotypes, reduction of heterogeneity, and extensive genetic mapping have all contributed to this progress. The rapid expansion of information from the human genome project will likely further accelerate this progress and assist in the discovery of susceptibility genes for schizophrenia. A greater understanding of disease mechanisms and the application of pharmacogenetics should also lead to improvements in therapeutic interventions. PMID- 11915954 TI - Toxic effects of benomyl on the ultrastructure during spermatogenesis of the earthworm Eisenia fetida. AB - The present study has investigated the toxic effect of benomyl on the ultrastructure of the male reproductive system and spermatozoa of the earthworm Eiseniafetida in a laboratory experiment. Three different concentrations of benomyl (8.3, 56, 112 mg/kg dry soil) were applied for one week. These applications caused abnormalities in ultrastructure of the cytophore, the spermatogonia, spermatids, and spermatozoa. The alterations include uncomplete forms of acrosomes, nuclear distortion, and disruption of microtubules. These micromorphological changes should be included in a model for predicting environmental hazards. PMID- 11915955 TI - Effect of dust on the viability of Vibrio fischeri in the Microtox Test. AB - The standard Microtox test involving the bioluminescent bacterium Vibrio fischeri is a frequently used ecotoxicological bioassay whose EC50, values have been correlated to acute toxicity parameters of vertebrates, to irritancy measures, and to cytotoxicity indices. The aims were to explore the dependence of light output on viable cell number, with the latter estimated with the naked eye using a colorimetric tetrazolium salt method, the effects of dust on the bioluminescence and cell viability, how the viability of the cells is affected after spills, and how spills can be sampled. The lower limit of the linear dynamic range of the light-emitting bacterium was first defined to be 3.7 x 10(7) cells/ mL, compared with 37 x 10(7) cells/mL in the Microtox assay. The effects of dust were then explored in the working range by the method of standard additions by adding 5-, 10-, and 20-mg amounts of Standard Reference Material Urban Dust 1649a. This simulated dust samples collected by a cordless vacuum technique involving a filter cassette. A mass of 20 mg dust totally inhibited the Microtox test at all times (5, 15, and 30 min). Masses of 5 and 10 mg dust lowered the luminescence significantly by 20 and 64%, respectively, after 30 min. However, the viability test was totally inhibited by 5 mg of dust. A spectrophotometric modification of the viability test using a wavelength of 508 nm was developed that was twice as sensitive as the naked eye test, and was as sensitive as the Microtox test. Mechanical shock involved with spilling and sampling bacterial reagent on hard surfaces killed the luminescent bacteria as shown by inhibition of luminescence. The optimum filter cassette for Microtox reagent collection was a 25-mm 1.00-microm PTFE filter in a 25-mm Delrin holder operated at 4.0 L/min, with a Tygon sampling probe. PMID- 11915956 TI - The effect of chromium on the hemoglobin concentration of Limnodrilus hoffmeisteri (Oligochaeta: Tubificidae). AB - The purpose of this study was to determine the toxic effects of polluted sediments, mainly chromium, the El Niagara reservoir (Aguascalientes, Mexico) on a benthic oligochaete species. Acute toxicity tests with hexavalent chromium in an artificial sediment-water system resulted in 24-, 48-, and 96-h LC50 values of 49.53, 22.81, and 5.11 mg available chromium/kg dry sediment, respectivley, in Limnodrilus hoffmeisteri. The uptake of chromium by tubificids from artificial and polluted reservoir sediments was found to increase with metal concentration in sediments and exposure time. The increase was higher in experiments with artificial sediment. Cr concentration in worms was related to hemoglobin content, which decreased significantly when Cr concentrations were above 1.0 microg/g dry weight. Bioavailable chromium in the El Niagara reservoir sediments may be an important factor limiting the benthic species in this ecosystem. PMID- 11915957 TI - Assessment of tolerant sunfish populations (Lepomis sp.) inhabiting selenium laden coal ash effluents. 1. Hematological and population level assessment. AB - Sunfish were collected from coal ash effluent-receiving streams and Ohio River watershed reference sites to assess the effects of exposure to low-level selenium concentrations. Selenium, copper, and arsenic concentrations were statistically higher in tissue samples from exposed fish than in reference fish. Leukopenia, lymphocytosis, and neutropenia were evident in exposed fish and were indicative of metal exposure and effect. White blood cell counts and percent lymphocyte values were significantly correlated with liver selenium concentrations. Plasma protein levels were significantly lower in exposed fish than in fish from the Ohio River, indicating that exposed fish may have been nutritionally stressed. Condition factors for fish from the ash pond-receiving streams were the same as, or lower than, those of fish from the reference sites. There was no evidence that the growth rate of fish in the receiving streams differed from that of fish in the reference streams. Despite liver selenium concentrations which exceeded reported toxicity thresholds and evidence of significant hematological changes, there were no significant differences in fish condition factors, liver-somatic indices, or length-weight regressions related to selenium. PMID- 11915958 TI - Assessment of tolerant sunfish populations (Lepomis sp.) inhabiting selenium laden coal ash effluents. 2. Tissue biochemistry evaluation. AB - Sunfish were collected from a fly ash pond-receiving stream and an Ohio River reference site to assess biochemical responses to coal ash effluent exposure. Selenium levels in sunfish from the receiving stream were higher than toxic thresholds associated with adverse population effects and reproductive impairment. Tissue biochemistry was found to be indicative of metal exposure and effect, but varied widely. Liver glycogen was positively correlated with increased liver metal levels, indicating no adverse effect upon stored carbohydrate levels. Lipid levels decreased with increasing metals, indicating possible nutritional stress. Protein levels increased with increasing metal levels, possibly due to the synthesis of proteins to sequester the metals. ATPase, dUTPase, and alkaline phosphatase activity generally decreased with exposure to ash pond metals, but remained within normal physiological ranges. Fish condition factors and liver somatic indices were correlated with liver lipid levels, dUTPase activity, and gill ATPase and alkaline phosphatase activity. Exposure to coal ash effluents produced biochemical markers of exposure that were associated with fish condition indicators; however, the indices themselves were not significantly affected by effluent exposure. PMID- 11915959 TI - Assessment of tolerant sunfish populations (Lepomis sp.) inhabiting selenium laden coal ash effluents. 3. Serum chemistry and fish health indicators. AB - Sunfish were collected from fly ash discharge-receiving streams to assess the possible effects of exposure to elevated selenium. Concentrations of selenium, copper, and arsenic were statistically higher in fish tissue (liver) samples from effluent-exposed fish than in reference fish. Several biomarkers were indicative of metal exposure and effect. Plasma protein levels and cholesterol levels were significantly lower in exposed fish, indicating nutritional stress. Ion levels (i.e., K) increased with exposure to ash pond metals, indicating possible gill damage. Fish from the receiving streams also had increased serum glucose and osmolality indicating possible acute stress due to sampling. Fish health assessments revealed a lower incidence of fin erosion, kidney discoloration, urolithiasis or nephrocalcinosis, liver discoloration, and parasites in exposed fish and a higher incidence of skin, eye, and gill aberrations. Condition factors of exposed fish were correlated with biomarker response and were the same as or lower than those of reference fish, but not related to selenium levels. Although several serum biochemical indicators differed between the ash pond-receiving stream and reference sites, pollutant exposure was apparently not sufficient to cause functional damage to critical organ systems. PMID- 11915960 TI - Can David and Goliath get along? Federal land in local places. AB - It is often unclear what the role of a local jurisdiction is with regard to land use management on nearby federal properties. Yet federal lands clearly impact nearby local communities. The US Department of Energy (DOE), with over 100 sites across the United States with varying degrees of environmental contamination, may be in a very difficult position with regard to relationships with local government about land use. Yet few, if any, studies have examined DOE land use issues. This study asks: (1) In general, how do local planners feel about federal government relationships with them? (2) Do local planners feel differently about the DOE than they do about other federal agencies? (3) What reasons explain any differences observed in answer to the second question? To answer these questions, local planners were interviewed from communities adjacent to non-DOE federal properties, and their responses compared to those of planners located near DOE facilities in the same regions. Findings showed that compared to other federal agencies that own land in the same regions, the DOE is relatively poorer at actively involving local officials in land use decisions at its sites. Primary reasons are the historic legacy of a culture of secrecy, focus on mission, and especially the lack of experience, training, or mandates in local planning cooperation. Findings also suggest that this attitude is markedly stronger in areas west of the Rocky Mountains. Recommendations for improved federal-local communications include the development of a vision for local government involvement that is supported by top levels of management and filtered effectively to the site level. PMID- 11915961 TI - Coping with change: the small experiment as a strategic approach to environmental sustainability. AB - The patterns of living, planning, and resource utilization as practiced in Western countries for decades are not sustainable; change of some sort is essential. But what changes are appropriate, and, perhaps more important, will people accept them? The small experiment framework provides a strategy for meeting the challenge of change. By encouraging participation, limiting the scale of initial change, and incorporating some aspects of the familiar, many of the difficulties that make change so problematic can be mitigated or even eliminated. An exploration, from a psychological perspective, of the characteristic difficulties surrounding potential change provides the context for a discussion of the components of the small experiment and an analysis of how these elements address these characteristic challenges. A comparison to adaptive management is drawn, and several concrete examples illustrate how the strategy has been used successfully to address a variety of environmental problems. PMID- 11915962 TI - Decline in endangered species as an indication of anthropic pressures: the case of European mink Mustela lutreola Western population. AB - Populations of threatened species, especially predators at the top of the food chain, may be affected by anthropic pressures. The endangered western population of European mink Mustela lutreola has shown a large decline over 50% of its natural range. M. lutreola disappeared from northwestern France between 1984 and 1997, and the decline was associated with an increase in mustelid trapping, changes in watercourse quality, and habitat modifications due to agricultural practices. The pattern of decline showed a fragmentation restricting the minks into very small areas. Trapping was the first known cause of mortality. Although feral American mink Mustela vison may compete with autochthonous carnivores, M. lutreola had disappeared from streams before the introduction of the American species, suggesting that competitive interactions were not responsible. Furthermore, American mink has never been found or has remained rare in 62.4% of the area from which M. lutreola has disappeared. During the past 25 years, permanent grassland surfaces were reduced by 40%, whereas fodder culture increased by 470%, causing considerable habitat changes. Furthermore, 55.7% of water courses were classified as being of bad quality or polluted. Therefore, our data suggests that a conjunction of intensive trapping, alterations in water quality and habitat modification was critical for the European mink's decline. Although there are difficulties in ascribing specific cause to distribution changes in a top predator, this decline can be regarded as an indication for anthropic pressures on natural habitats. PMID- 11915963 TI - Modeling urban growth effects on surface runoff with the integration of remote sensing and GIS. AB - A methodology is developed to relate urban growth studies to distributed hydrological modeling using an integrated approach of remote sensing and GIS. This linkage is possible because both studies share land-use and land-cover data. Landsat Thematic Mapper data are utilized to detect urban land-cover changes. GIS analyses are then conducted to examine the changing spatial patterns of urban growth. The integration of remote sensing and GIS is applied to automate the estimation of surface runoff based on the Soil Conservation Service model. Impacts of urban growth on surface runoff and the rainfall-runoff relationship are examined by linking the two modeling results with spatial analysis techniques. This methodology is applied to the Zhujiang Delta of southern China, where dramatic urban growth has occurred over the past two decades, and the rampant urban growth has created severe problems in water resources management. The results revealed a notably uneven spatial pattern of urban growth and an increase of 8.10 mm in annual runoff depth during the 1989-1997 period. An area that experienced more urban growth had a greater potential for increasing annual surface runoff. Highly urbanized areas were more prone to flooding. Urbanization lowered potential maximum storage, and thus increased runoff coefficient values. PMID- 11915964 TI - Sedimentary causes and management of two principal environmental problems in the lower Yellow River. AB - Flood and water shortage are two of the leading environmental problems around the world, and among the causes of the problems is sedimentation. The Yellow River brought disastrous floods in its lower reaches in Chinese history. Today, although floods caused by the river are still a formidable hazard hanging over China, it cannot provide the lower reaches with enough usable water. The ineradicable flood hazard and newly emerged water shortage problems of the river are proved to be closely associated with its immense sediment load. The over loaded flow of the river can quickly fill the reservoirs and unceasingly raise the riverbed, attenuating the capacity of reservoirs to suppress floods and provide more water for dry seasons and of river channels to convey floods. Also, the high sediment content pollutes the water and reduces the volume of usable water. In virtue of the intimate linkage between these problems and the formidable sediment load in the river, the solution to these problems should be based on sedimentation management. After reviewing the defects and merits of management measures implemented and proposed, a management scenario composed of multiple measures are recommended. Beside of persistent soil conservation to reduce the huge sediment load, more reservoirs to check sediment and regulate river flows, approaches to alleviating riverbed accretion, interbasin water transfer to mitigate water deficiency, and so on, an emphasis should be laid on use of muddy flows in order to scatter the sediment in a vast area, which was a natural process but has been interrupted by construction of embankments. PMID- 11915965 TI - Design and performance of a channel reconstruction project in a coastal California gravel-bed stream. AB - A 0.9 km-reach of Uvas Creek, California, was reconstructed as a sinuous, meandering channel in November 1995. In February 1996, this new channel washed out. We reviewed project documents to determine the basis for the project design and conducted our own historical geomorphological study to understand the processes operating in the catchment and project reach. The project was designed using a popular stream classification system, based on which the designers assumed that a "C4" channel (a meandering gravel-bed channel) would be stable at the site. Our historical geomorphological analysis showed that the reach had been braided historically, typical of streams draining the Franciscan Formation in the California Coast Ranges, with episodic flows and high sand and gravel transport. After the project washed out, Uvas Creek reestablished an irregular, braided sand and-gravel channel, although the channel here was narrower than it had been historically, probably due to such factors as incision caused by gravel mining. Our study casts doubt on several assumptions common in many stream restoration projects: that channel stability is always an appropriate goal; that channel forms are determined by flows with return periods of about 1.5 years; that a channel classification system is an easy, appropriate basis for channel design; and that a new channel form can be imposed without addressing the processes that determine channel form. PMID- 11915966 TI - Demographic analysis of tree colonization in a 20-year-old right-of-way. AB - Past tree colonization dynamics of a powerline-right-of-way (ROW) corridor in the Haut-Saint-Laurent region of Quebec was studied based on the present age distribution of its tree populations. This colonization study spans 20 years, from 1977 (ROW clearance) to 1996. The sampled quadrats were classified into six vegetation types. Tree colonization dynamics were interpreted in each type, and three distinct patterns were identified. (1) Communities adapted to acidic conditions were heavily colonized by Acer rubrum, at least for the last 12 years. (2) Communities adapted to mesic or to hydric conditions were more intensely colonized in the period 1985-1987 than in the following 9 years; this past success in tree colonization may have been caused by herbicide treatments, which could have facilitated tree establishment by damaging the herbaceous and shrub vegetation. (3) Cattail, vine-raspberry, and reed-dominated communities contained few tree individuals, with almost all trees establishing between 1979 and 1990; those three vegetation types appear as the most resistant to tree invasion in the ROW studied. This study supports the need for an integrated approach in ROW vegetation management, in which the selection of vegetation treatment methods would depend on the tree colonization dynamics in each vegetation type. Minimizing disturbances inflicted on ROW herbaceous and shrub covers should be the central strategy because disturbances jeopardize natural resistance to future tree invasion, except in communities adapted to acidic conditions where the existing vegetation does not prevent invasion by A. rubrum. Many trees are surviving the successive cutting operations by producing new sprouts each time, particularly in communities adapted to mesic and hydric conditions. In these cases, mechanical cutting should be replaced by a one-time stump-killing operation, to avoid repeated and unsuccessful treatments of the same individuals over time. PMID- 11915967 TI - Changing farmers' land management practices in the hills of Nepal. AB - This paper sheds light on changing farmers' land management practices in two mountain watersheds, with and without extemal assistance, in the western hills of Nepal. Information used in the analysis were obtained through a survey of 300 households, group discussion, key informant interviews, and field observation conducted during April-September 1999. Confronted with ever-decreasing landholding size due to a steadily growing population and scarcity of nonfarming employment opportunities, farmers in both watersheds have increasingly adopted assorted types of structural and biological measures to control soil erosion, landslides, gully expansion, and soil nutrient loss to maintain or even enhance land productivity. Adoption of guly control measures, construction of the retention walls, alley cropping, use of vegetative measures for landslide control, mulching, and use of green manure and chemical fertilizers are found significantly high in the project area due to the provision of technical and financial support, whereas composting is found significantly high in the nonproject area. Different from the traditionally held beliefs, population pressure on a finite land resource has brought positive change in land management. However, the experience from both watersheds indicates that there is limit to the extent that resource poor farmers can respond to land degradation without any extemal assistance. Required is the arrangement for appropriate polices and support services and facilities enabling farmers to adopt locationally suitable and economically attractive land management technologies. PMID- 11915968 TI - The concept of habitat diversity between and within ecosystems applied to river side-arm restoration. AB - Since returning an ecosystem to its pristine state may not be realistic in every situation, the concept of habitat diversity is proposed to help decision-makers in defining realistic restoration objectives. In order to maintain habitat diversity and enhance the long-term success of restoration, process-oriented projects should be preferred to species-oriented ones. Because the hydrogeomorphological processes that influence biodiversity operate at different spatiotemporal scales, three scales are considered: river sectors, floodplain waterbodies, and mesohabitats within each waterbody. Based on a bibliographical review, three major driving forces are proposed for incorporation into the design of restoration projects: (1) flow velocity and flood disturbances, (2) hydrological connectivity, and (3) water supply. On the sector scale, increased habitat diversity between waterbodies can be achieved by combining various intensities of these driving forces. On the waterbody scale, increased habitat diversity within the ecosystem can be achieved by varying water depth, velocity, and substrate. The concept is applied to a Rhjne River sector (France) where three terrestrialized side arms will be restored. Two were designed to be flood scoured, one having an additional supply of groundwater, the other being connected to the river at both ends. The third cannot be scoured by floods because of upstream construction and would be supplied by river backflow through a downstream connection. Habitat diversity within the ecosystem is exemplified on one side arm through the design of a sinuous pathway combined with variation of water depth, wetted width, and substrate grain size. Self-colonization of the side arms is expected owing to the restoration of connectivity to upstream sources of potential colonizers. PMID- 11915969 TI - Managing urban trees and their soil envelopes in a contiguously developed city environment. AB - Urban Hong Kong is covered by high building, road, and population densities. Its urban morphology is inherently not conducive to extensive or high-grade greening. Recent renewal of old areas has squeezed out some limited interstitial plantable space, although in new development areas modest spaces have been earmarked for greenery. The study aims at evaluating the major constraints to urban trees and their companion urban soil envelopes and at providing specific recommendations to improve tree management in the city. The analysis covers the above-ground confinements that dampen tree performance, the less tangible but rather difficult institutional restrictions that impose a somewhat unnecessary lid on tree planting, the multiplicity of players and stakeholders involved in urban-tree management that militates against coordination and cooperation, the widespread occupation of underground space by utility lines often to the exclusion of trees, and the extremely poor quality of urban soils that are often used without amelioration to support tree growth. The management recommendations furnish practical suggestions and hints to improve the short- and long-term welfare of trees in terms of quality, quantity, and spatial distribution. The conclusion enumerates some concrete measures for consideration by decision-makers to upgrade the city's greenery to close the gap between science and policy. PMID- 11915970 TI - Evaluating microbial indicators of environmental condition in Oregon rivers. AB - Traditional bacterial indicators used in public health to assess water quality and the Biolog system were evaluated to compare their response to biological, chemical, and physical habitat indicators of stream condition both within the state of Oregon and among ecoregion aggregates (Coast Range, Willamette Valley, Cascades, and eastern Oregon). Forty-three randomly selected Oregon river sites were sampled during the summer in 1997 and 1998. The public health indicators included heterotrophic plate counts (HPC), total coliforms (TC), fecal coliforms (FC) and Escherichia coli (EC). Statewide, HPC correlated strongly with physical habitat (elevation, riparian complexity, % canopy presence, and indices of agriculture, pavement, road, pasture, and total disturbance) and chemistry (pH, dissolved O2, specific conductance, acid-neutralizing capacity, dissolved organic carbon, total N, total P, SiO2, and SO4). FC and EC were significantly correlated generally with the river chemistry indicators. TC bacteria significantly correlated with riparian complexity, road disturbance, dissolved O2, and SiO2 and FC. Analyzing the sites by ecoregion, eastern Oregon was characterized by high HPC, FC, EC, nutrient loads, and indices of human disturbance, whereas the Cascades ecoregion had correspondingly low counts of these indicators. The Coast Range and Willamette Valley presented inconsistent indicator patterns that are more difficult to characterize. Attempts to distinguish between ecoregions with the Biolog system were not successful, nor did a statistical pattern emerge between the first five principle components and the other environmental indicators. Our research suggests that some traditional public health microbial indicators may be useful in measuring the environmental condition of lotic systems. PMID- 11915971 TI - Application of cost-effectiveness analysis to multiple products: a practical guide. AB - The appropriate interpretation of cost-effectiveness results and its use in treatment decision making have been debated vigorously. In this report we have summarized the decision rules described in the health economics literature to determine which intervention should be chosen from among multiple treatment options for a given disease, using a graphic framework. The implications of different means of expressing budget constraints on treatment choice are examined. PMID- 11915972 TI - Diagnostic testing or empirical neuraminidase inhibitor therapy for patients with influenza-like illness: what a difference a day makes. AB - OBJECTIVE: To determine the clinical and economic trade-offs of available diagnostic and treatment options for patients with suspected influenza infection. DESIGN: Decision analytic model. PATIENTS: A simulated cohort of adults with influenza-like illness of less than 48 hours' duration. INTERVENTIONS: Patients received (1) no testing or treatment; (2) rapid testing for influenza, with neuraminidase inhibitors (NIs) prescribed for individuals who test positive; or (3) empirical NI therapy. A validated prediction rule to determine the probability of influenza infection based on patient signs and symptoms is available. The monetary value attributable to the incremental benefits of NI therapy must be quantified during the clinician visit. OUTCOME: Incremental cost per case of influenza treated with NIs. RESULTS: The decision whether to treat, test, or prescribe NIs empirically for patients presenting with influenza-like illness of less than 48 hours' duration depends on the probability of influenza and the value of the benefits of NI therapy. If the incremental value of NI therapy exceeds $200 and the probability of influenza is greater than 35%, empiric NI therapy is the preferred option. The test option has a very narrow band of preference due to suboptimal test characteristics and cost. CONCLUSION: The decision whether to prescribe NIs based on clinical criteria or restrict NI use to patients with a confirmed laboratory diagnosis of influenza depends on the likelihood of infection, the accuracy and cost of the diagnostic test, and the benefits attributable to NI therapy. Clinicians must consider these factors when determining cost-effective use of NIs. PMID- 11915973 TI - Health-related quality of life in early rheumatoid arthritis: impact of disease and treatment response. AB - OBJECTIVE: To document the burden of early rheumatoid arthritis (RA) on health related quality of life (HQL) and compare changes in HQL across 2 treatments. STUDY DESIGN: Analysis of HQL scores among patients enrolled in a multicenter, double-blind, randomized control trial of early RA treatment. PATIENTS AND METHODS: A total of 424 patients with early RA were randomized to 1 of 2 treatment groups: etanercept or methotrexate. Patients were treated and followed for 52 weeks. Health-related quality of life was assessed before and throughout treatment using the Medical Outcomes Study Short Form 36 Health Survey (SF-36) and the Health Assessment Questionnaire (HAQ). The HQL burden of RA was established by comparing SF-36 scale scores to general US population norms. The impact of treatment on HQL was determined by comparing scores on both SF-36 and HAQ scales. RESULTS: Before treatment, RA patients showed significant decrements in scores on all SF-36 scales and summary measures in comparison with age- and sex-matched general US population norms, multivariate analysis of variance (MANOVA) F(8,2815) = 204.6, P < .0001. After 52 weeks of treatment, 7 of 8 SF-36 scales and the physical summary measure remained significantly below the general US population norm, MANOVA F(8,2815) = 41.9, P < .0001. Patients randomized to etanercept showed significantly better HQL improvement earlier in treatment than patients randomized to methotrexate on the SF-36 physical summary, MANOVA F(10,4230) = 6.1, P< .0001, the SF-36 arthritis-specific health index, MANOVA F(10,4230) = 8.5, P < .0001, and the HAQ, MANOVA F(10,4230) = 14.7, P < .0001. At 52 weeks, there were no significant differences between treatment groups. CONCLUSIONS: Rheumatoid arthritis places tremendous disease burden on patients' HQL. Successful treatment of early RA improved HQL. Etanercept showed a rapid HQL response. PMID- 11915974 TI - Changes in related drug class utilization after market withdrawal of cisapride. AB - BACKGROUND: Recent Food and Drug Administration-mandated and company-initiated withdrawals of drug products from the marketplace have had an impact on utilization in related drug classes. OBJECTIVE: To investigate the impact of withdrawal of the prokinetic agent cisapride (Propulsid) on utilization of other gastrointestinal (GI) agents. STUDY DESIGN: A longitudinal, retrospective study using electronic prescription data from a state-funded geriatric prescription benefit program. PATIENTS AND METHODS: Prescription claims for 2644 patients using cisapride between January 10, 2000, and October 1, 2000, were analyzed with respect to points in time at which (1) prospective drug utilization review edits were implemented denying reimbursement of cisapride because of drug interactions, (2) the manufacturer announced its intent to cease production, and (3) the agent was withdrawn from the market. Prevalence of use, claims volume, and expenditures were compared for cisapride, proton pump inhibitors, histamine-2 receptor antagonists, and the prokinetic agent metoclopramide during these periods. RESULTS: Use of cisapride decreased precipitately even before implementation of a "medical exception only" reimbursement policy. After the change in policy, metoclopramide use increased, although this increase was not proportional to cisapride's decline. Although total GI expenditures declined within the cisapride cohort, this change had little impact on overall program GI expenditures. CONCLUSIONS: The loss of cisapride did not significantly affect program-wide costs for GI drugs. However, the withdrawal of cisapride appears to have resulted in increased use of metoclopramide, a medication with a more serious adverse effect profile than cisapride. Further study is needed to evaluate the long-term clinical impact of such therapy changes. PMID- 11915975 TI - Development and maintenance of a community-based hepatitis C registry. AB - OBJECTIVE: To develop a model for community-population- or health system-based registries of all patients with diagnosed hepatitis C, to facilitate clinical care and epidemiologic studies. STUDY DESIGN: Geographically defined, population based cohort study. METHODS: Registry subjects were identified using January 1, 1990, to December 31, 1999, data from the Rochester Epidemiology Project (REP), which lists all diagnoses for Olmsted County residents recorded by clinicians during visits to Olmsted County medical providers. We supplemented diagnostic data with information from laboratory databases that record all hepatitis C testing in Olmsted County. All diagnoses based on the REP and laboratory databases were confirmed by medical record review. Proposed data elements to be included in a hepatitis C registry were identified and defined, and data collection methodology was tested. RESULTS: A total of 355 subjects (62% male) were identified in the total community population of 130,000. Both the diagnostic summary database (n = 309, 87%) and the laboratory database (n = 46, 133%) were important in the identification of subjects for the registry. Nine additional subjects with diagnostic or laboratory evidence of hepatitis C refused the legislatively mandated (Minnesota statute) medical records research authorization and could not be included in the registry. Most desired data elements were available in the medical records. CONCLUSIONS: Both medical visit diagnostic summaries (administrative or billing data) and laboratory databases are required to identify subjects with physician-based diagnoses of hepatitis C. Few patients refused the authorization required for inclusion in a research registry. PMID- 11915976 TI - Value for money in health and medical care services. PMID- 11915977 TI - Ventricular pacing in the presence of tricuspid valve disease. AB - Pacemaker implantation following tricuspid valve surgery remains challenging, but recent developments in lead technology have significantly improved the options for ventricular pacing. In the presence of significant tricuspid regurgitation, steroid-eluting active-fixation leads should be used routinely, and in patients with a tricuspid prosthesis, the use of a dedicated cardiac venous pacing system is likely to be the best option. PMID- 11915978 TI - Conduction properties of the crista terminalis and its influence on the right atrial activation sequence in patients with typical atrial flutter. AB - The conduction properties of the crista terminalis (CT) and its influence on the right atrial activation sequence were analyzed in 14 patients with typical atrial flutter (AF). Atrial mapping was performed with 35 points of the right atrium during typical AF and during atrial pacing performed after linear ablation of inferior vena cava-tricuspid annulus (IVC-TA) isthmus. Atrial pacing was delivered from the septal isthmus at cycle lengths of 600 ms and the tachycardia cycle length (TCL). The right atrial activation sequence and the conduction interval (CI) from the septal to lateral portion of the IVC-TA isthmus were analyzed. During AF, the conduction block line (CBL) (detected by the appearance of double potentials along the CT and craniocaudal activation on the side anterior to CT) was observed along the CT in all patients. The TCL and CI during AF were 254 +/- 19 and 207 +/- 14 ms, respectively. During pacing at a cycle length of 600 ms, the CBL was observed along the CT in four patients, however, a short-circuiting activation across the CT was observed in the remaining ten patients. The CI during pacing at 600 ms was 134 +/- 38 ms, shorter than that during AF (P < .0001). During pacing at the TCL, the CBL was observed along the CT in all patients. The presence of the CBL along the CT prevented a short circuiting activation across the CT and resulted in the same right atrial activation as observed during AF. With the formation of the CBL, the CI significantly increased to 206 +/- 17 ms and was not different from that during AF. These data suggest that the conduction block along the CT is functional. It was presumed that presence of conduction block at the CT has some relevance to the initiation of typical AF though it was not confirmed. PMID- 11915979 TI - Alternans in QRS amplitude during ventricular tachycardia. AB - Although the value of T wave alternans as an index of electrical instability has been extensively investigated, little is known about QRS alternans during VT. Intracardiac electrograms of 111 episodes of spontaneous monomorphic regular VT retrieved from implantable defibrillators in 25 patients were retrospectively selected. Three beat series, representing the total amplitudes and amplitudes from baseline to summit and from baseline to lower point of 16 or 32 successive QRS complexes before deliverance of electrical therapy were generated for each episode. Spectral analysis was then performed using the fast Fourier transform. VT was considered as alternans if the magnitude of the spectral power at the 0.5 cycle/beat frequency was greater than the mean +/- 3 SD of the noise in at least one of the three spectral curves. QRS alternans was present in 23 (20%) of 111 episodes and in 9 (36%) of 25 patients. Alternans was not related to the VT cycle length, QRS duration, QRS amplitude, signal amplification, nor to clinical variables. Alternans was more frequently detected in unipolar configuration and when a higher number of complexes was included in analysis. Failure of antitachycardia pacing was more frequent in case of alternans VT (50% vs 75% success in nonalternans VT, P = 0.05). Spontaneous termination before deliverance of therapy occurred in 16 non-alternans VT but never in alternans episodes (P = 0.02). Alternans in QRS amplitude is a relatively common finding during VT and could be associated with failure of antitachycardia pacing and lack of spontaneous termination. Lower efficacy of electrical therapies in case of QRS alternans must be confirmed in a way to improve the effectiveness of antitachycardia pacing. PMID- 11915980 TI - Adjustment of maximum automatic sensitivity (automatic gain control) reduces inappropriate therapies in patients with implantable cardioverter defibrillators. AB - Patients with ICDs might experience oversensing associated with inappropriate shock therapy when paced at low bradycardia rates or if they have a low intrinsic rhythm. The amplifier gain of automatic gain control is maximal at long RR intervals and might lead to oversensing of myopotentials. The hypothesis was that an individual adjustment of the sensitivity floor could reduce inappropriate ICD therapies. Fourteen patients implanted with the VENTAK MINI I/II and III in whom oversensing and/or inappropriate shocks had been documented were included in this evaluation based on stored episodes. Sensitivity was modified in all ICDs by means of noninvasive software downloading that allowed reprogramming of the maximum sensitivity from 0.14 mV in two steps to 0.27 mV ("Normal," "Less," "Least"). Provocation testing (deep inspiration, sit-ups, and abdominal pressing) was conducted at "Nominal" settings (0.14 mV) while the intracardiac electrogram with annotated event marker was running continuously and was repeated at 0.22 mV (Less) and 0.27 mV (Least). All patients evaluated had documented spontaneous episodes due to oversensing before the sensitivity floor was reprogrammed. During provocation testing at Nominal settings, oversensing could be documented in 9 of (64.3%) 14 patients. Provocation testing was repeated after the sensitivity selection was reprogrammed and oversensing could not be provoked in any of the 14 implanted devices. Sensitivity was reprogrammed in all 14 patients to a higher value (5 patients Less, 9 patients Least). After sensitivity adjustment, VF was induced in all patients to verify appropriate arrhythmia detection and termination. Sensitivity reprogramming in patients with documented oversensing eliminates the incidence of inappropriate shock therapy without compromising the ability to detect VF appropriately. PMID- 11915981 TI - How to test mode switching in pacemakers implanted in patients: the MOST study. AB - Optimal management of atrial arrhythmias with dual chamber pacemakers requires proper performance of automatic mode switching (AMS). The aim of this study was to develop a reliable technique to test the AMS function by using an external electronic device capable of mimicking the occurrence of supraventricular arrhythmias (Supraventricular Arrhythmia Simulator [SAS]). The SAS delivers low voltage pulse trains (200 mV, 20 ms) through two skin electrodes. Each pulse train lasts 15 seconds and starts synchronously with a pacing pulse of the implanted pacemaker to avoid interference from the operator. The pulse train rate is set at 350, 250, and 160 beats/min to simulate AF, atrial flutter, and atrial tachycardia (AT), respectively. Thirty-five patients implanted with Vitatron pacemakers, whose AMS system has been previously validated, were enrolled. Atrial and ventricular sensing were programmed in unipolar mode at 0.5 mV and in bipolar mode at > 2 mV, respectively. All pulses from the SAS were detected by the atrial channel at an amplitude ranging from 1 to 3 mV. The test proved to be safe and reliable at rest and during exercise. AMS occurred immediately at onset or at offset of atrial arrhythmias, and no adverse interference on pacemaker function was seen from the SAS. In conclusion, the described technique and the SAS are safe and reliable for patient and pacemaker function and can be proposed as a useful method to verify proper performance of AMS function irrespective of the type of implanted devices. PMID- 11915982 TI - Magnetocardiographic and magnetic resonance imaging for noninvasive localization of ventricular arrhythmia origin in a model of nonischemic cardiomyopathy. AB - Ventricular arrhythmia may in myocardial failure arise as a consequence of remodeling related to hypertrophy and dilatation. In surgically repaired tetralogy of Fallot (TOF), which carries a substantial risk for ventricular arrhythmias and sudden death, the situation is even more complex and several potential arrhythmia mechanisms exist. The authors wanted to test a completely noninvasive localization technique, magnetocardiography (MCG) and magnetic resonance imaging (MRI), to define the origin of ventricular ectopic beats (VEBs) in this model of nonischemic cardiomyopathy. The study included 84 patients with surgically repaired TOF, all 11 subjects with VEBs Lown grade > or = 2 on a 24 hour Holter were included. From 37-channel MCG registrations, the underlying current dipole was computed for the VEBs, and for anatomic correlation MRIs were produced. Eight patients had VEBs of altogether 11 morphologies during the MCG recording. The RVOT was the origin of 6 VEBs, while 4 originated from the RV free wall, and 1 VEB could not be localized. Applying this completely noninvasive technique, it was possible to define different origins of RV ectopies in a complex heart model. Thus, VEBs originated from the RVOT, suggesting a relation to surgical scars and from the nonoperated parts of the RV, supporting that ventricular remodeling might be of equal importance for arrhythmogenesis in this model of cardiomyopathy. This technique can assist in answering the important question of whether there is any difference in prognostic information depending on the origin and related mechanism of VEBs in this and other high risk conditions. PMID- 11915983 TI - Characterization of a long linear thermolesion in rabbit atria by high density mapping. AB - A major problem in radiofrequency ablation is to evaluate if linear atrial lesions are transmural and continuous. Characteristics of the atrial electrograms recorded from these lesions might be useful to determine the completeness of linear atrial lesions. In seven isolated perfused rabbit atria, a long transmural linear lesion was made with a hot needle (perfused with water of 80 degrees C) (width 2 +/- 0.2 mm). The lesion extended from the orifice of the superior caval vein to the A V ring. High density mapping (240 electrodes, 7.5 x 7.5 mm) of the right atrium was performed before and after the lesion. Complete bidirectional conduction block was confirmed by pacing close at either side of the lesion. No change in conduction or electrogram characteristics occurred outside the lesion. From the center of the lesion, low amplitude double potentials were recorded. The conduction delay around the lesion and the width of the double potentials showed a high correlation (R2 = 0.99) and were both dependent of the site of pacing. From the boundaries of the lesion towards its center, the amplitude and slope of the unipolar electrograms decreased exponentially by 72 +/- 5 and 85 +/- 3%. From the decay of these electrotonic potentials a space constant (lambda of 0.79 +/- 0.04 mm) could be calculated for the lesion. Unipolar electrograms recorded from a complete and transmural linear atrial lesion are electrotonic in nature. Their characteristics could be used to evaluate the width and depth of the lesion. PMID- 11915985 TI - Variability of Holter electrocardiographic findings in patients fulfilling the noninvasive MADIT criteria. Multicenter Automatic Defibrillator Implantation Trial. AB - In the MADIT study, a selected group of postinfarction patients with asymptomatic nonsustained ventricular tachycardia (NSVT) has been shown to benefit from prophylactic ICD treatment. The present study analyzed the variability of NSVT in a patient population fulfilling the non-invasive MADIT criteria. Three consecutive Holter ECGs were performed in weekly intervals in 68 postinfarction patients with an LVEF < or = 0.35. Patients with NSVT underwent programmed ventricular stimulation (PVS); patients were implanted with an ICD if sustained VT or VF was inducible. If NSVT was found in at least two recordings, the arrhythmia was defined as reproducible. In 28 (41%) of the 68 patients, NSVT was found in at least one recording. Seventeen patients revealed NSVT in the first, the remaining 11 in the second registration; no patient had NSVT only in the third Holter. Of the patients with NSVT, 50% had only one, 39% had two, and 11% had three positive recordings. Thus, reproducible NSVT was found in only 50% of the patients with NSVT. Predictors for reproducibility were LVEF > 0.27, NYHA Class I, absence of digitalis therapy, and > 2 NSVT per 24-hour period. Reproducible NSVT was not associated with risk factors such as elevated mean heart rate, reduced heart rate variability, late potentials, or inducibility of sustained VT during PVS. During 17 +/- 9 months of follow-up, seven (10%) patients experienced arrhythmic events: two without and five with previously documented NSVT. In the latter patients, first occurrence of NSVT was consistently in the first Holter; only two of them had reproducible NSVT. In postinfarction patients, the risk factor NSVT exhibits marked spontaneous variability, especially in those with a low number of NSVT per 24-hour period, LVEF < 0.27 or NYHA III, which limits its clinical value as a selection criterion for PVS. Reproducibility of NSVT itself does not seem to be an independent risk factor. PMID- 11915984 TI - Ventricular coupling of electrical and mechanical dyssynchronization in heart failure patients. AB - We studied the relationships of electrical and mechanical synchronization in patients with heart failure (CHF) and various degree of ventricular conduction delays. Ninety-two CHF patients (60 +/- 13 years old, LVEF < 45%), NYHA II-III IV, and 35 age-matched control subjects were studied with angioscintigraphic phase analysis. We measured ejection fractions (LVEF, RVEF) and calculated the total activation time for the left (TtLV) and right ventricle (TtRV), and the synchronization time between right and left ventricle (TRVLV), and between LV apex and base (Tab). Patients were divided into three groups according to QRS duration: group 1 < 120 ms (n = 28), group 2 < 150 ms (n = 23), group 3 > or = 150 ms (n = 41). In group 1: LVEF = 31.1 +/- 10.9%, RVEF = 30.1 +/- 12.6%, TtLV = 204 +/- 70 ms, TtRV = 183 +/- 61 ms, TRVLV = 7 +/- 33 ms, Tab = 29 +/- 23 ms. In group 2, these were: 27.8 +/- 9.1%, 27.8 +/- 8.8%, 227 +/- 95 ms, 248 +/- 137 ms, 35 +/- 42 ms*, and 39 +/- 53 ms respectively. In group 3: LVEF = 20.5 +/- 9.5%t, RVEF = 28.4 +/- 16.1%, TtLV = 304 +/- 155 mst, TtRV = 234 +/- 106 mst, TRVLV = 64 +/- 42 mst, and Tab = 67 +/- 48 ms*, all P < 0.001 versus controls *P < 0.05 versus G1, tP < or = 0.01 versus G1. A significant relation links QRS to both inter- and intraventricular asynchrony (TRVLV: r = 0.65; TtLL: r = 0.70, Tab: r = 0.60), and to LV function (r = 0.72); while LVEF relates more closely to intraventricular asynchrony: TtLV (r = 0.52), TtLL (r = 0.67), than to interventricular asynchrony: TRVLV (r = 0.48); P < 0.01, P < or = 0.001. In CHF patients, electromechanical and contractile alterations are coupled; regional activation may be an early parameter allowing the detection of ventricular dyssynchronization. PMID- 11915986 TI - Rapid recovery of cardiac function after catheter ablation of persistent junctional reciprocating tachycardia in children. AB - Four children (age range 3 weeks to 15 years) with tachycardia induced cardiomyopathy due to permanent junctional reciprocating tachycardia (PJRT) underwent successful catheter ablation of the tachyarrhythmia circuit without any complications during or after the procedure. In all four patients, the left ventricular fractional shortening improved to normal within 2 weeks of the procedure. These results argue for a more aggressive approach to the therapy of PJRT when associated with tachycardiomyopathy at a young age. PMID- 11915987 TI - Optimal atrioventricular delay setting determined by QT sensor of implanted DDDR pacemaker. AB - QT interval (QTI) may change when cardiac function is improved by optimizing the AV delay. QTI is used as the sensor for rate responsive pacemakers. Evoked (e)QTI is measured as the time duration from the ventricular pace-pulse to the T sense point, which is the steepest point of the intracardiac T wave. The relationship between AV delay and eQTI and cardiac function was studied in 13 patients (74.2 +/- 9.3 [SD] years old) with an implanted QT-driven DDDR pacemaker. A special pacemaker software module was downloaded into the pacemaker memory for eQTI data logging. AV delay was set at 100, 120, 150, 180, 210, and 240 ms. Cardiac output (CO) was measured by continuous Doppler echocardiography. eQTI was 343.3 +/- 22.4, 345.1 +/- 22.5, and 343.4 +/- 23.2 ms (P < 0.01, repeated ANOVA) and CO was 4.2 +/- 0.8, 4.6 +/- 0.8, and 4.2 +/- 0.8 L/min (P < 0.0001, repeated ANOVA) when AV delay was set at the AV delay shortened by one step (AV[-]) and prolonged by one step (AV[+]) from the AV delay at which QT interval was maximum (AV[max]) in seven patients, in whom the peak AV delay at which the eQTI was maximal could be identified. eQTI decreased from 341.1 +/- 20.9 to 339.4 +/- 21.1 ms (P < 0.0001) and CO decreased from 4.4 +/- 1.4 to 4.1 +/- 1.3 L/min (P < 0.005) when AV delay was prolonged from AV(max) to AV(+) in all patients. eQTI decreased from 345.1 +/ 22.5 to 343.3 +/- 22.4 ms (P < 0.0005) and CO decreased from 4.6 +/- 0.8 to 4.2 +/- 0.8 L/min (P < 0.05) when AV delay was shortened from AV(max) to AV(-) in seven patients. Thus, CO was maximal when AV delay was set at the AV delay at which eQTI was maximal. In conclusion, the optimal AV delay can be predicted from the eQTI sensed by an implanted pacemaker, and automatic setting of the optimal AV delay can be achieved by the QT sensor of an implanted pacemaker. PMID- 11915989 TI - Narrow QRS tachycardia that accelerates spontaneously. What is the explanation? PMID- 11915988 TI - The challenge of endocardial right ventricular pacing in patients with a tricuspid annuloplasty ring and severe tricuspid regurgitation. AB - On occasion, patients with a tricuspid annuloplasty ring may require permanent cardiac pacing. Although it is technically possible to pass a ventricular transvenous lead through a tricuspid valve with an annuloplasty ring, the procedure is complicated by considerable chamber enlargement and mechanical distortion of the tricuspid valve often with severe residual tricuspid regurgitation. Over a 25-month period, transvenous ventricular lead placement following insertion of a tricuspid annuloplasty ring was successfully performed in five patients (three women). The patient mean age was 66 years (range 55-77 years). Four cases had slow atrial fibrillation and another paroxysmal atrial fibrillation requiring His-bundle ablation. Two patients had mitral valve replacement and two aortic and mitral valve replacements. All patients had residual severe to torrential tricuspid regurgitation. Seven ventricular steroid eluting screw-in leads were used. Single leads were used in three cases, whereas in two others, two ventricular leads were attached to a dual chamber pulse generator. Although technically difficult, ventricular lead placement was successful using standard guidewires with broad curvatures. Satisfactory acute and follow-up stimulation thresholds and sensing were obtained with the only complication being an intraoperative lead dislodgement, prompting a second ventricular lead. Successful transvenous lead placement across a tricuspid annuloplasty ring is possible. PMID- 11915990 TI - The imprecision in heart rate correction may lead to artificial observations of drug induced QT interval changes. AB - Because of the known limitations of the Bazett and other heart rate correction formulas, it has been proposed that studies of drug induced QT interval changes should use several different heart rate correction formulas and that the consistency of findings by a majority of such formulas should be considered as valid. The aim of this article was to show that such an approach is inappropriate. Using the database of the EMIAT trial, data of QT and RR intervals were taken from electrocardiograms of the first postrandomization visit of 1,402 patients. Of these, 309 were on amiodarone and beta-blockers, 395 on amiodarone and off beta-blockers, 318 on beta-blockers and off amiodarone, and 380 off amiodarone and off beta-blockers. An investigation of drug induced QT interval changes was modeled by evaluating the corrected QT (QTc) interval differences between patients on and off amiodarone, and on and off beta-blockers. A set of 31 previously published heart rate correction formulas was used. In addition to calculating the QTc difference between on and off drug for each formula, the success of heart rate correction was judged by computing correlation coefficients between QTc and RR intervals (ideally corrected QTc values should be independent of heart rate). The difference between on and off drug QT intervals was also evaluated by logarithmic regression models between uncorrected QT and RR intervals in data taken from patients on and off treatment. The QTc interval prolongation on amiodarone was confirmed by all heart rate correction formulas but the extent of the prolongation differed from formula to formula and ranged from 13.6 to 30.9 ms. Of the 31 formulas, 3 reported QTc interval shortening on beta-blockers (up to -11.8 ms) and 28 reported QTc interval prolongation (up to +16.8 ms). The distribution of the results provided by the different formulas suggested that beta-blocker treatment led to a QTc interval prolongation by approximately 7 ms (e.g., +7.4 ms by the Fridericia formula, P = 0.002). The on treatment QTc changes obtained by different formulas were closely correlated to their correction success. Formulas that provided QTc intervals almost independent of the RR intervals estimated approximately 20 ms QTc prolongation on amiodarone and no QTc change on beta-blockers. QT/RR regression analysis confirmed that while amiodarone led to substantial QT prolongation, there was no change of QT interval on beta-blockers beyond the change in heart rate. The study showed that the concept of "majority voting" by different heart rate correction formulas is inappropriate and may lead to erroneous conclusions. PMID- 11915991 TI - Restored atrial excitability after late recanalization in a patient with atrial standstill and acute myocardial infarction. AB - Atrial standstill is electrophysiologically characterized by the loss of spontaneous excitation in atrial muscle and the inability to cause action potential firing upon electrical stimulation. Clinical diagnosis of transient standstill of the right atrium was made in a patient with acute occlusion of the right coronary artery and acute renal failure. Percutaneous coronary intervention, performed 5 days after the onset, restored the coronary blood flow and resulted in full recovery of electrical activity and regular sinus rhythm. PMID- 11915992 TI - Runaway pulse generator malfunction resulting from undetected battery depletion. AB - Runaway pacemaker is an uncommon, potentially lethal circuit malfunction characterized by sudden onset of erratic pacing at rapid nonphysiological rates. Two patients with a single chamber pacemaker (Medtronic ST 8331 and 8419) presented with episodic dizziness. ECG revealed recurrent decrescendo amplitude episodes of runaway stimuli at 2,400 and 2,600 ppm, approximately 3 seconds in duration, separated by pacing at 62.5 and 65 ppm, respectively. Fortunately the runaway stimuli were subthreshold and did not result in capture of the ventricle. Emergency pulse generator replacement was uneventful. Both leads were normal and both pulse generators had low battery voltages at 1.488 and 1.78 V, respectively. PMID- 11915993 TI - An operative case of inferior vena cava stenosis due to fibrosis around permanent pacemaker leads. AB - A 56-year-old woman presented with general fatigue, dyspnea on exercise, and hepatomegaly subsequent to secondary implantation of a pacemaker. On admission, cardiac echo examination showed tricuspid valve regurgitation due to a migrated looped lead. At surgery, there was evidence of severe stenosis at both orifices of the superior and inferior vena cavae due to fibrous tissue around the leads. We removed the fibrous tissue, pacing leads, and generator. New leads were anchored onto the epicardium and a generator was inserted under the rectus muscle. The postoperative course was satisfactory and the symptoms disappeared. PMID- 11915994 TI - Severe mitral regurgitation with right ventricular pacing, successfully treated with left ventricular pacing. AB - A case of severe mitral regurgitation with refractory heart failure, after atrioventricular junction ablation and pacemaker implant, was solved with left ventricular pacing. Mitral regurgitation was related to a change in segmental left ventricular motion during right ventricular pacing. PMID- 11915995 TI - Magnesium deficiency during lactation as a precipitant of ventricular tachyarrhythmias. AB - A young ICD recipient with a history of syncope and idiopathic polymorphic ventricular tachycardia/ventricular fibrillation presented after an ICD discharge. She had delivered her first child 8 days prior to the event and she had been lactating. Numerous short runs of polymorphic ventricular tachycardia/ventricular fibrillation resolved with aggressive replacement of magnesium and elimination of breast-feeding. PMID- 11915996 TI - Alternate pacing sites for patients with tricuspid valve prostheses. AB - The objective of this study was to pace via a coronary vein to avoid interfering with the tricuspid valve prosthesis function. Pacing leads were inserted into the posterior cardiac vein in a 68-year-old woman (patient 1), and in the great cardiac vein and the right auricle in a 32-year-old woman (patient 2). In patient 1 the stimulation threshold was 1.8 V at implant and stabilized at 3.0 V at the 24-month follow-up. In patient 2 the ventricular pacing threshold was 1.2 V at implant and stabilized at 0.7 V at the 24-month follow-up. The cardiac output at rest increased 43% during atrioventricular synchronous pacing compared to ventricular pacing. Long-term stable ventricular pacing via the coronary venous system was obtained. PMID- 11915998 TI - Utility of right precordial leads at higher intercostal space positions to diagnose Brugada syndrome. AB - We encountered a patient with Brugada syndrome whose body surface ECG revealed that the specific ST-segment elevation was found only at higher positions of leads V1 and V2, above the third intercostal space. Only these right precordial leads at higher intercostal space positions were useful to identify Brugada syndrome in this case. PMID- 11915997 TI - Implantation of a single chamber pacemaker in patients with triple mechanical valve prosthesis: utilization of coronary sinus distal branches to stimulate the left ventricle. AB - The implantation of a permanent pacemaker and its relative leads is still a challenge in patients with mechanical tricuspid valve prosthesis. The implant is usually performed epicardially or during valve replacement to avoid any damage to the valve and/or early damages to the lead. We describe the case of a patient with a tricuspid valve prosthesis implanted with a permanent single-chamber pacemaker using an endocardial lead positioned in a distal branch of the coronary sinus to stimulate the left ventricle. PMID- 11915999 TI - Pacing in the middle cardiac vein in a patient with tricuspid prosthesis. AB - A man with acute endocarditis developed complete heart block several days after the tricuspid and aortic valve replacement. Several weeks after implantation, his epimyocardial pacing leads developed a high threshold and failed to capture the ventricle at the maximal pulse width and output of the pacemaker. An angled-tip lead was placed in the middle cardiac vein for ventricular pacing. The pacing and sensing thresholds of this lead were within the expected range during follow-up. Therefore, in patients with prosthetic tricuspid valve, pacing in the middle cardiac vein should be considered before open-chest placement of the epimyocardial lead. PMID- 11916000 TI - Georges Weiss' fundamental law of electrostimulation is 100 years old. AB - In 1901, Georges Weiss published a voluminous paper that was the result of the charge by the Commission International du Parc-aux-Prince to investigate whether there are measures to make mutually comparable and to classify the different devices that physiologists used for nerve and muscle stimulation. Georges Weiss was born August 26, 1859 in Bischweiler (Alsace, France). He trained as an engineer in Paris and afterward began his medical training and received his medical doctorate in 1889. In the same year he was appointed "Professeur Agrege, " and "Preparateur" at the Department of Medical Physics of the Medical Faculty at Paris. He made many contributions to physiology, but his main field of interest was electrophysiology. At the end of the nineteenth century the measuring capabilities for electrical stimulation pulses were limited and stimulation theories were based more on speculation than on measurements. Weiss found a fascinating method to produce short-lasting pulses of defined amplitude and duration. He constructed bridges by conducting threads within a circuit that were then destroyed by an air rifle bullet driven by liquid carbonic acid to produce short-lasting pulses. To investigate double pulses, the measuring system was expanded in the same manner, now with four bridging threads. The experiments were carried out with remarkable accuracy. The results included: (1) the threshold quantity that is the voltage-time-product, is a linear function of the pulse duration; (2) there is always a minimum of the delivered energy dependent on pulse duration; (3) pulse shape plays no role in electrostimulation. The physiologists were not so impressed by the Weiss report as it did not really meet the requirements as expressed by the title. There was no measuring technology available at that time to measure the quantity of devices with short-lasting pulses in the millisecond range. No wonder that the importance of the findings was not really perceived by the scientific community. It can be concluded that if there are statements in the literature contradicting one of the above three Weiss theorems, one can infer that the investigation is questionable. PMID- 11916002 TI - The revised NASPE/BPEG generic code for antibradycardia, adaptive-rate, and multisite pacing. North American Society of Pacing and Electrophysiology/British Pacing and Electrophysiology Group. AB - In light of evolving pacemaker technology and increasing interest in multisite pacing, the Committee on the Development of Position Statements (CDPS) of the North American Society of Pacing and Electrophysiology (NASPE) created an ad hoc Pacemaker Mode Code Task Force in April 2001 under the chairmanship of David L. Hayes, MD, for the purpose of bringing the NASPE/British Pacing and Electrophysiology Group (BPEG) Generic Pacemaker Code (NBG Code) up to date. The task force, whose members are the authors of this article, designed a revised NBG Code in which three major issues were taken into account. First, it was recognized that all modern pacemakerpulse generators are capable of extensive bidirectional communication with an external programming device, making them "communicating" pulse generators as defined by the current (1987) NBG Code. Second, it was decided that a means of providing basic information regarding the location of multisite pacing would be a useful ingredient of the NBG Code. Third, in view of the extensive antibradycardia pacing capabilities common in modern implantable cardioverter defibrillators and the availability of the NASPE/BPEG Defibrillator Code (NBD Code), it was considered unnecessary for the NBG Code to address the presence or absence of antitachycardia features. The resulting updated version of the NBG Code as described herein was endorsed by the BPEG on September 20, 2001 and adopted by the NASPE Board of Trustees on October 18, 2001. The structure of the revised NBG Code differs from that of the previous version in two respects alone: Position IV specifies only the presence or absence of rate modulation, and Position V specifies only the location or absence of multisite pacing (i.e., biatrial or biventricular pacing with at least two stimulation sites in each case) more than one stimulation site in any single cardiac chamber, or any combination of these. The revised NBG Code is deliberately configured to avoid confusion with earlier mode codes, and it is the authors' hope that it will serve as an enhanced resource for communication among those engaged in every phase of the multidisciplinary practice of cardiac rhythm management. PMID- 11916001 TI - New antiarrhythmic drugs for the treatment of atrial fibrillation. PMID- 11916003 TI - Molecular analysis of mutations and polymorphisms of the Lewis secretor type alpha(1,2)-fucosyltransferase gene reveals that Taiwan aborigines are of Austronesian derivation. AB - The origins of the Taiwan aborigines have not been fully resolved. Anthropological and linguistic studies have indicated that their ancestry is mainly Austronesian or Malayopolynesian. Some polymorphisms and mutations in the secretor type alpha(1,2)-fucosyltransferase (FUT2) gene are found in Taiwan aborigines. In this study, we analyzed the frequency of eight mutations and one polymorphism of the FUT2 gene in Taiwan aborigines and other ethnic groups in order to explore the origins of these groups. The results showed that the C302T, G428A, and fusion gene mutations were specific for Thai, Caucasians, and Japanese, respectively. The A385T mutation was specific for Asians including Taiwan aborigines. The genetic frequencies of C571T were much higher in Taiwan aborigines (1.96% to 20.4%), Filipinos (13.2%), and Indonesians (3.30%) as compared with Thai (0.57%), Chinese (0.65% to 1.12%), Japanese (0%), and Caucasians (0%). The frequencies of the G849A mutation were also higher in Taiwan aborigines (0.38% to 21.57%), Filipinos (6.80%), and Indonesians (1.48%) than in Thai (0.94%), Chinese (0-0.37%), Japanese (0%), or Caucasians (0%). Deletion of a 3-bp region (nt 688 to nt 690) was found only in Filipinos (0.85%), Indonesians (0.74%), and three tribes (0.42% to 2.70%) of Taiwan aborigines, but not in other populations. The C628T mutation was found only in Taiwanese Han, Thai, and two tribes of Taiwan aborigines. The genetic frequency of the C357T polymorphism was much higher in Asians than in Caucasians. The genetic analysis confirms that the origins of Taiwan aborigines are Austronesian and that they are closely related to the Filipino and Indonesian populations. We suggest that mutations or polymorphisms of the FUT2 gene are very good markers for investigating population genetics. PMID- 11916004 TI - Identification of mutations in the arginine vasopressin receptor 2 gene causing nephrogenic diabetes insipidus in Chinese patients. AB - Congenital nephrogenic diabetes insipidus (NDI) is, in most instances, a rare X linked recessive renal disorder (MIM 304800) characterized by the clinical symptoms of polyuria, polydipsia, and dehydration. The X-linked NDI is associated with mutations of the arginine vasopressin receptor type 2 (AVPR2) gene, which results in resistance to the antidiuretic action of arginine vasopressin (AVP) in the renal tubules and collecting ducts. Identification of mutations in the AVPR2 gene can facilitate early diagnosis of NDI, which can prevent serious complications such as growth retardation and mental retardation. We analyzed three unrelated Chinese NDI families and identified three mutations: R106C, F287L, and R337X. In addition, an A/G polymorphism at cDNA nucleotide position 927 (codon 309L) was identified. A functional expression assay of the R106C and F287L mutants in COS-7 cells revealed that both mutants show significant dysfunction and accumulate intracellular cyclic adenosine monophosphate in response to AVP hormone stimulation. These results facilitate the diagnosis of NDI at the molecular level in the Chinese population, and provide insight into the molecular pathology of NDI. PMID- 11916005 TI - Thirteen single-nucleotide polymorphisms (SNPs) in the alcohol dehydrogenase 4 (ADH4) gene locus. AB - The human alcohol dehydrogenase 4 (ADH4) gene encodes the class II ADH4 pi subunit, which contributes to the metabolization of a wide variety of substrates, including ethanol, retinol, other aliphatic alcohols, hydroxysteroids, and lipid peroxidation products. Here we report the results of systematic screening for single-nucleotide polymorphisms (SNPs) in the ADH4 gene by means of direct sequencing combined with a polymerase chain reaction method. A total of 16 genetic variations including 13 SNPs were found; 4 in the 5' flanking region, 4 in the 5' untranslated region, and 8 within introns. No variation was found in coding, 3' untranslated, or 3' flanking regions. Eight of the 13 SNPs were not reported in the NCBI dbSNP database or any previous publications. Our SNP map presented here should provide tools to evaluate the role of ADH4 in complex genetic diseases and a variety of pharmacogenetic effects. PMID- 11916006 TI - Nonaka myopathy is caused by mutations in the UDP-N-acetylglucosamine-2 epimerase/N-acetylmannosamine kinase gene (GNE). AB - This is the first report on mutations of the UDP-N-acetylglucosamine-2 epimerase/N-acetylmannosamine kinase gene (GNE) in Nonaka myopathy or distal myopathy with rimmed vacuoles (OMIM 605820), an autosomal recessive neuromuscular disorder. Sequence and haplotype analyses of GNE in two siblings with Nonaka myopathy from a Japanese family revealed that both patients were compound heterozygotes for a C-->T transition (A460V) in exon 8 and a G-->C transition (V572L) in exon 10. Their parents and a normal elder brother were all carriers for one or the other of the mutations. GNE mutations are known to cause two other disorders: sialuria (OMIM #269921) and autosomal recessive inclusion body myopathy (IBM2, OMIM #600737). Mutations associated with sialuria are located in the epimerase domain, and those associated with IBM2 are in the epimerase or the kinase domain or both, whereas the mutations we observed in the Nonaka myopathy patients were located in the sugar kinase domain of the gene. Thus, Nonaka myopathy is the third disease known to be caused by GNE mutations. PMID- 11916007 TI - Eight novel mutations and functional impairments of the LDL receptor in familial hypercholesterolemia in the north of Japan. AB - In the course of investigations of familial coronary artery disease in Hokkaido, the northland of Japan, we identified 13 families affected by familial hypercholesterolemia. Among them, we identified eight novel mutations of the low density lipoprotein (LDL) receptor gene, four of which caused frameshifts: (1) a 7-bp deletion at nucleotide (nt) 578-584 (codon 172-174, exon 4); (2) a 14-bp insertion at 682nt (codon 207-208, exon 4); (3) a 49-bp deletion at nt 943-991 (codon 294-310, exon 7); and (4) a one-base insertion of C to a stretch of C3 at nucleotides 1687-1689 or codon 542. The others included (5) a T-to-C transition at nt 1072 causing substitution of Cys for Arg at codon 337 (C337R, exon 8); (6) a splice-site G-to-T substitution in intron 11; (7) a splice-site G-to-C substitution in intron 11; and (8) a G-to-T transition at nt 1731 causing substitution of Trp for Cys at codon 556 (W556C, exon 12). To disclose the functional consequences of novel mutations, we characterized each of these mutations by two assays in peripheral lymphocytes, i.e., uptake of fluorescently labeled LDL by LDL receptors, and measurement of cell surface-bound LDL receptor protein using specific monoclonal antibody against LDL receptor. PMID- 11916008 TI - Functional polymorphism in the promoter region of the gelatinase B gene in relation to coronary artery disease and restenosis after percutaneous coronary intervention. AB - The matrix metalloproteinases appear to play an important role in the development and progression of atherosclerotic lesions. We studied the C-1562T polymorphism of the gelatinase B promoter in relation to coronary artery disease and restenosis after a percutaneous coronary intervention (PCI) in Koreans. To determine the frequency of the C-1562T allele, we examined 63 patients with coronary artery disease who underwent both PCI and 6-month follow-up coronary angiograms (CAGs), and 67 control patients with a normal CAG with respect to their clinical data and genotype. Frequencies of the C/C homozygotes and the non C/C heterozygotes and homozygotes (C/T and T/T) were 94% and 6% in the normal CAG group, and 76.2% and 23.8% in the patient group, respectively. This gave a relative risk of 0.203 (95% CI: 0.063-0.651, P = 0.005) for coronary artery disease when the C/C genotype was compared with the non-C/C genotype. In the patient groups, the allele frequencies of the C/C and non-C/C were 80% and 20% in the nonrestenotic subgroup, and 71.4% and 28.6% in the restenotic subgroup (P = 0.554). No T/T homozygote was found in any of the groups. We conclude that C/C homozygosity is a potential genetic protective factor for coronary artery disease in Koreans. PMID- 11916009 TI - Distinctive distribution of AIM1 polymorphism among major human populations with different skin color. AB - The genetic background for human skin color has been a major topic in human genetics; however, its molecular basis is still unclear. The gene for the AIM-1 protein (AIM1) was recently found to be responsible for the body color of medaka fish. In the search for the genes controlling human skin color variations, we have investigated genetic polymorphisms of this gene, and we have found a single nucleotide polymorphism that has clear association with major human populations in terms of skin color. PMID- 11916010 TI - A new haplogroup pattern displayed in Fujian Han in China. AB - Human Y-chromosomal binary polymorphisms have been considered to preserve the paternal genetic legacy and provide evidence on human evolution and the genetic relationships among and demographic history of different populations. To reveal the genetic origin and immigration of the Fujian Han, 13 binary markers on the Y chromosome were used to screen Fujian Han by allele-specific polymerase chain reaction. The results indicated that the M9G marker was highly prevalent (96.20%), suggesting a significant genetic drift. In addition, M122C frequency was only 22.78%, and M45A and M103T were default. The distinctive haplogroup frequencies (H1, H5, and H6/7/8) imply that the haplogroup pattern is a relatively ancestral and interim type. PMID- 11916011 TI - Evidence for an association between plasma platelet-activating factor acetylhydrolase deficiency and increased risk of childhood atopic asthma. AB - Platelet-activating factor (PAF), which has been implicated in the pathophysiology of inflammation in asthma, is degraded and inactivated by PAF acetylhydrolase (PAFAH). Approximately 4% of the Japanese population lacks plasma PAFAH due to a loss-of-function variant (Val279Phe) in the PAFAH gene. Although lack of PAFAH activity is thought to be a risk factor for asthma, there are conflicting findings concerning association between the Val279Phe variant and asthma. In this study, we conducted transmission disequilibrium tests of 118 Japanese parent-child trios identified through mite-sensitive atopic asthmatic children. A case-control study was also carried out. The Phe279/Phe279 genotype was found more frequently in children with atopic asthma (13%) than in their parents (6%) or in controls (4%). Results of the genotypic transmission test were significant, and the Phe279/Phe279 genotype was transmitted preferentially to asthmatic children. Our data support an association between deficiency in PAFAH activity and atopic asthma. PMID- 11916012 TI - Enantiomer analysis of a new street drug, 3,4-methylenedioxy-N-methyl-butanamine, in rat urine. AB - A new street drug, 3,4-methylenedioxy-N-methyl-butanamine (MBDB), has been found in Japan recently. The stereoisomer monitoring and the urinary excretion kinetics are not determined in biological fluids even though abused MBDB is a racemic form [enantiomer ratio (-/+) = 1.00]. The present studies were done by high performance liquid chromatography (HPLC) equipped with a chiral activity column at 40 degrees C using urine specimens from five Wistar rats. Urine samples were collected over six time intervals after a single oral administration of racemic MBDB (30 mg/kg). Unchanged MBDB and 3,4-methylenedioxybutanamine (BDB), an N demethylated metabolite, were found in the rats' urine. Each enantiomer of MBDB and BDB was monitored (peak resolution > 1.00) by HPLC analysis within 30 min. For both MBDB and BDB, the (+)-isomers were excreted a little more than the (-) isomers. The stereoselective disposition of BDB was more remarkable than that of MBDB and was observed in the urine throughout the study (p < 0.05). The urinary excretion of MBDB showed significant difference between the two enantiomers from 4 to 20 h (p < 0.05). The amount of MBDB excreted up to 24 h was 34.7+/-2.8% of the administered dose: 17.6+/-1.4% for (+)-isomer and 17.1+/-1.5% for (-)-isomer. The amount of BDB was 4.9+/-1.0%; 2.9+/-0.6% for (+)-isomer and 2.0+/-0.4% for ( )-isomer. The enantiomer ratio (-/+) of MBDB and BDB was 1.00 or a little smaller. The ratio (-/+) of MBDB changed from 1.00+/-0.02 to 0.88+/-0.09 by 24 h, and that of BDB from 0.68+/-0.03 to 0.78+/-0.02. The ratio (-/+) for MBDB and BDB accumulated up to 24 h was 0.97+/-0.01 and 0.70+/-0.06, respectively, and the total ratio (-/+) of the two substances was 0.93+/-0.02 (p < 0.05). These findings suggested that the stereoselective disposition of racemic MBDB was different from that of 3,4-dimethylenedioxyamphetamine and 3,4 dimethylenedioxymethamphetamine and was similar to that of methamphetamine. PMID- 11916013 TI - Prednisone concentrations in human hair. PMID- 11916014 TI - Toxicological analysis of chlorhexidine in human serum using HPLC on a polymer coated ODS column. AB - A simple and reliable high-performance liquid chromatographic (HPLC) method for analyzing chlorhexidine in human serum was developed. After the addition of an internal standard, levomepromazine, 0.2 mL serum was deproteinized with 10% perchloric acid. The acidic supernatant was neutralized with 1M potassium carbonate solution, and the insoluble salt was removed by centrifugation. An aliquot of the supernatant was applied to HPLC with UV detection (260 nm). HPLC separation was achieved on a polymer-coated ODS column equilibrated with acetonitrile/water containing 0.05% trifluoroacetic acid, 0.05% heptafluorobutyric acid, and 0.1% triethylamine (40:60, v/v). The calibration curve was linear in the concentration range from 0.05 to 50.0 microg/mL, and the lower limit of detection was 0.05 microg/mL. The accuracy and precision of the method were evaluated at concentrations of 0.5 microg/mL and 5.0 microg/mL. The coefficients of variation ranged from 4.0 to 4.5%. The concentration of chlorhexidine in the serum of a patient who died after a suspected intravenous injection of chlorhexidine gluconate was determined. PMID- 11916015 TI - Blood concentration of milnacipran in a case of a fatal automobile accident. AB - A fatal automobile accident involving milnacipran and ethyl alcohol is reported. The drug was identified and quantitated in blood by high-performance liquid chromatography-diode-array detection. The concentration of milnacipran in peripheral blood was 3.15 microg/mL exceeded 10.5 times the therapeutic concentration. The blood ethyl alcohol level was 1.3 g/L. PMID- 11916016 TI - In vivo metabolism of 4-bromo-2,5-dimethoxyphenethylamine (2C-B) in the rat: identification of urinary metabolites. AB - The in vivo metabolism of 4-bromo-2,5-dimethoxyphenethylamine (2C-B), a ring substituted psychoactive phenethylamine, in the rat was studied. Male Wistar rats were administered 10 mg/kg of 2C-B hydrochloride orally, and 24 h urine fractions were collected. After enzymatic hydrolysis of the urine samples, the metabolites were extracted by liquid-liquid extraction and analyzed by gas chromatography mass spectrometry. 2-(4-Bromo-2,5-dimethoxyphenyl)-ethanol, 4-bromo-2,5 dimethoxyphenylacetic acid, 2-(2-hydroxy-4-bromo-5-methoxyphenyl)-ethylamine, 2 (2-methoxy-4-bromo-5-hydroxyphenyl)-ethylamine, 1-acetoamino-2-(2-hydroxy-4-bromo 5-methoxyphenyl)-ethane, and 1-acetoamino-2-(2-methoxy-4-bromo-5-hydroxyphenyl) ethane were identified as 2C-B metabolites. These findings suggest that at least two metabolic pathways for 2C-B are operative in rats. The first pathway leads to the corresponding aldehyde metabolite by deamination, which is subsequently reduced or oxidized, to give the corresponding alcohol and carboxylic acid metabolites. The second pathway leads to the corresponding 2-O-desmethyl or 5-O desmethyl metabolites in which the amino group is subsequently acetylated. PMID- 11916017 TI - Determination of methemoglobin and total hemoglobin in toxicological studies by derivative spectrophotometry. AB - A method for the determination of methemoglobin in the presence of other hemoglobin subforms (i.e., oxy-, deoxy-, and carboxyhemoglobin) by use of derivative spectrophotometry is proposed. The method, which uses the first derivative of the spectrum at 645 nm, is straightforward and expeditious, so it is of special interest to forensic toxicology laboratories as it allows the simultaneous determination of the methemoglobin saturation percentage and the hemoglobin concentration. This facilitates interpretation of the results and provides a better understanding of the significance of methemoglobin saturation in specific cases. Based on an analysis of interferences, the presence of other hemoglobin subforms or of endogenous components of plasma does not detract in any way from the performance of the method. PMID- 11916018 TI - Influence of Helix pomatia enzyme preparations on the oxidative conversion of some clostebol acetate metabolites in urine. AB - Clostebol acetate (4-chloro-testosterone acetate) is an anabolic steroid used for fattening purposes in cattle breeding. To safeguard public health, its use has been prohibited by the European Commission since 1986. Screening for its urinary metabolites is therefore an important tool for the control of possible violations. Because those metabolites appear conjugated to glucuronic acid or sulfate, deconjugation prior to analysis is necessary. This work describes the variability in results seen with the use of various commercial preparations of Helix pomatia (SHP) for enzymatic hydrolysis of the conjugates. A simultaneous oxidative side reaction was observed, converting metabolites with a 3-OH-4-ene structure into a 3-oxo-4-ene structure. This was not observed when samples were incubated without enzyme or in the presence of heat-inactivated SHP. GC-MS analysis revealed oxidation of some metabolites of clostebol acetate, 4-chloro-4 androsten-3alpha-ol-17-one and 4-chloro-4-androsten-3alpha,17beta-diol, changing them into other metabolites, 4-chloro-4-androsten-3,1 7-dione and clostebol (4 chloro-testosterone), respectively. Based on the difference in cross-reactivities of the antibodies for these metabolites, comparative analysis in enzyme immunoassay, following enzymatic hydrolysis, confirmed this transformation. This oxidative conversion phenomenon could be of great importance when considering the choice or target analytes for screening bovine urine. PMID- 11916019 TI - Detection of morphine in blood and urine samples from horses administered poppy seeds and morphine sulfate orally. AB - The objective of this study was to determine if the administration of poppy seeds to horses would result in detectable concentrations of morphine in urine and blood samples, as has been shown to occur in humans. In this study blood and urine samples were collected following administration of poppy seeds and morphine sulfate orally to four horses. Urine samples were subjected to enzyme-linked immunosorbent assay (ELISA) for the presence of morphine. All urine samples testing positive by ELISA, as well as plasma samples collected after administration of the 10-g doses of poppy seeds, were analyzed by gas chromatography-mass spectrometry for the presence of morphine. Morphine was detectable in the plasma samples for at least 4 h after administration of 10 g of poppy seeds. Morphine was detectable in urine samples for up to 24 h after administration of 10 g, 5 g, and 1 g of poppy seeds and 426.7 microg of morphine as morphine sulfate. The results of this study indicate that horses that consume or are administered poppy seeds may have detectable concentrations of morphine in their urine and plasma for hours after administration. PMID- 11916020 TI - Simultaneous determination of eleven benzodiazepine hypnotics and eleven relevant metabolites in urine by column-switching liquid chromatography-mass spectrometry. AB - An automated column-switching liquid chromatographic-atmospheric pressure chemical ionization-mass spectrometric (LC-APCI-MS) method for the simultaneous determination of frequently encountered benzodiazepine hypnotics and their relevant metabolites in urine has been established. A column-switching online solid-phase extraction technique was employed to enhance sensitivity and eliminate tedious sample pretreatment. The combination of an N-vinylacetamide containing hydrophilic polymer online extraction column and a C18 semi-micro LC column provided the successful separations and MS determinations of the 11 benzodiazepines and 11 relevant metabolites tested in this study. The detection limits ranged 2-10 ng/mL and 10-50 ng/mL in the SIM and full-scan modes, respectively, using 50 microL of urine. The calibration curves were linear up to 500 ng/mL for all these analytes in the SIM mode. The present method provided successful forensic applications for the proof of benzodiazepine administration. PMID- 11916022 TI - Making the most of practice abilities--the 2002 VPMA conference. Veterinary Practice Management Association. PMID- 11916021 TI - Multiresidue determination of endosulfan and metabolic derivatives in human adipose tissue using automated liquid chromatographic cleanup and gas chromatographic analysis. AB - A multiresidue method based on normal-phase liquid chromatography (LC) cleanup and gas chromatography-electron capture detection-mass selective detection (GC ECD-MSD) analysis has been developed for the determination of endosulfan and its main metabolic derivatives in human adipose tissues. Analytes were extracted by dissolving the fat samples in n-hexane, and the hexanic extracts were directly injected onto the silicagel column of the automated LC cleanup system. Purified LC extracts were analyzed by GC-ECD or GC-MSD, without any solvent exchanges or preconcentration steps. The high efficiency of the high-performance liquid chromatographic cleanup for the elimination of fats allowed to reach detection limits for all analytes at low nanograms-per-gram concentration levels. The optimized overall analytical procedure was applied to 18 selected human mammary adipose and abdominal fat tissue samples. p,p'-DDE, hexachlorobenzene, and beta HCH were the most frequently detected compounds, and residues of endosulfan sulfate and -ether were also found in several samples. All findings were confirmed by an additional GC-MS-MS analysis of the LC sample extracts. PMID- 11916023 TI - Survey of the stocking densities at which sheep are transported commercially in the United Kingdom. AB - A total of 6578 sheep carried in 74 vehicles to one slaughter plant were monitored, and stocking densities, in terms of m2 floor area available per 100 kg liveweight, were estimated from the dimensions of the vehicle pens and estimates of liveweight based on chest girth measurements. The observed stocking densities ranged from 0.29 to 2.00 m2/100 kg liveweight, and the average density was 0.65 m2/100 kg. One per cent of the sheep were carried at estimated stocking densities of less than 0.3 m2/100 kg, 37 per cent at densities between 0.3 and 0.6 m2/100 kg and 57 per cent at densities between 0.6 and 0.9 m2/100 kg. Over 30 per cent of the animals in the survey were transported at densities higher than the working recommendations made by the Farm Animal Welfare Council. PMID- 11916024 TI - Analysis of the variations in clinical signs shown by 254 cases of equine headshaking. AB - A national survey of headshaking in 254 horses was undertaken to describe the clinical signs of the condition as observed by horse owners. Principal component analysis was used to determine the underlying structure of 11 signs and the criteria by which the affected horses could be most effectively differentiated; the analysis suggested five components with a variance greater than one which together explained over 60 per cent of the total variance. Other analyses of the data indicated that headshaking could develop at any age and that twice as many males were affected as females; 64 per cent of the horses shook their heads seasonally and geldings were more likely than mares to be seasonally affected. Seasonal headshaking tended to be significantly worse on sunny days but improved on rainy days, windy days, at night and indoors. PMID- 11916025 TI - Serum concentrations of acute phase proteins in dogs with leishmaniasis. AB - The concentrations of haptoglobin, C-reactive protein and ceruloplasmin were measured in symptomatic and asymptomatic dogs naturally infected by Leishmania infantum, and in healthy uninfected dogs to determine the potential value of these proteins for the diagnosis and prognosis of leishmaniasis. The concentrations of the acute phase proteins were significantly higher in the dogs with leishmaniasis than in the control dogs, and the concentration of C-reactive protein was significantly higher in the symptomatic dogs than in the asymptomatic dogs. There were no correlations between the acute phase proteins and the gamma globulins, the albumin/globulin ratio or the titre of anti-leishmanial antibodies. PMID- 11916026 TI - Usefulness of systemic inflammatory response syndrome criteria as an index for prognosis judgement. PMID- 11916027 TI - Interlocking nail repair of a fractured femur in a turkey. PMID- 11916028 TI - Leptospirosis in Turkey. PMID- 11916029 TI - Granulomatous orchitis associated with Histoplasma-like organisms in porcupines (Erethizon dorsatum). PMID- 11916031 TI - Oral lesions in cattle. PMID- 11916030 TI - NDFAS and herd health plans. National Dairy Farm Assured Scheme. PMID- 11916032 TI - Reproduction of PMWS with a 1993 Swedish isolate of PCV-2. PMID- 11916033 TI - Bare marketing. PMID- 11916034 TI - CDNA microarray assessment for ozone-stressed Arabidopsis thaliana. AB - Various detrimental factors in the environment damage plants, resulting in growth inhibition or withering. However, it is not easy to identify causal factors by visually inspecting the damaged plants. Therefore, we have developed a sensitive and reliable method for plant diagnosis, based on measuring changes in expression of a set of genes in a DNA microarray. With this method, we have been able to detect and discriminate between plants stressed by ozone, drought, or wounding. PMID- 11916035 TI - Assessing the environmental fate of chemicals of emerging concern: a case study of the polybrominated diphenyl ethers. AB - It is suggested that assessments of chemicals of emerging concern can be rationally structured around a multistage process in which fate and risk are evaluated with increasing accuracy as new data become available. An initial tentative and approximate assessment of fate and risk can identify key data gaps and justify and direct further investigations, which progressively improve the reliability of the assessment. This approach is demonstrated for a class of chemicals, the polybrominated diphenyl ethers (PBDEs), which is of increasing concern, but about which there is presently a lack of comprehensive data on properties, sources, fate and effects. Specifically, 20 PBDE congeners are investigated using the suggested approach and research needs are identified. PMID- 11916036 TI - Time series study of concentrations of SO4(2-) and H+ in precipitation and soil waters in Norway. AB - Along with a steady reduction of acid inputs during 14 years of intensive forest monitoring in Norway, the influence of acid deposition upon soil water acidity is gradually reduced in favour of other and internal sources of H+ and sulphate, in particular from processes in the upper soil layer. We used statistical analyses in two steps for precipitation, throughfall and soil water at 5, 15 and 40 cm depths. Firstly, we employed time series analyses to model the temporal variation as a long-term linear trend and a monthly variation, and by this filtered out residual, weekly variation. Secondly, we used the parameter estimates and the residuals from this to show that the long term, the monthly and the weekly variation in one layer were correlated to similar temporal variation in the above, adjacent layer. This was strongly evident for throughfall correlated to precipitation, but much weaker for soil water. Continued acidification in soil water on many plots suggests that the combined effects of anthropogenic and natural acid inputs exceed in places the buffering capacity of the soil. PMID- 11916037 TI - Organochlorine pesticide contamination in grassland-nesting passerines that breed in North America. AB - Organochlorine pesticides and metabolites were measured in grassland-nesting passerines that breed in North America. We also examined testes of male birds for abnormalities that may have resulted from pesticide exposure. Forty-four of 99 individuals contained one or more organochlorine pesticides above the detection limit, representing nine of 10 species. The most prevalent compound detected was p,p'-DDE (minimum-maximum levels: 7.55-285.85 ng/g, carcass concentration). Insectivorous birds had significantly higher levels of p,p'-DDE than both omnivores and granivores. Birds that frequented moist grassland habitats had significantly higher levels of p,p'-DDE than those that frequented drier grassland habitats. No evidence of feminization was observed in any of the testes analyzed, however, other endpoint effects of contamination (e.g. hormone levels and immunological parameters) should be investigated in future studies. PMID- 11916038 TI - Phytotoxicity of dredged sediment from urban canal as land application. AB - Phytotoxicity of dredged sediment from Hangzhou section of the Grand Canal as land application was evaluated by pakchoi (Brassica chinensis L.) germination tests and pot experiments. Germination rates of pakchoi in the dredged sediment and in sediment-applied soils were both significantly higher than that in the soil controls, while the germination rate between the sediment-applied soils was no significant difference. In pot experiments, plant height and biomass were increased by the dredged sediment application rate in the rate of lower than 540 t ha(-1), but decreased when the application rate was over this rate. Concentrations of Zn and Cu in pakchoi were linearly increased with the increasing of the application rate of the dredged sediment. Both plant height and biomass of pakchoi in sediment-treated red soil were higher than that in sediment treated paddy soil, regardless the application rate. The results suggest that plant biomass of pakchoi may be used as an indicator of the phytotoxicity of the dredged sediment. It also showed that red soil is more suitable to accept the dredged sediment than paddy soil, and 270 t ha(-1) is a safe application rate both in red soil and paddy soil. PMID- 11916039 TI - Wild rats as monitors of environmental lead contamination in the urban area of Milan, Italy. AB - Twenty-one wild brown rats (Rattus norvegicus) were collected from four areas of Milan (Italy). Main organs were harvested from each rat, fixed in 10% neutral buffer formalin, and embedded in paraffin. Organ sections were stained with hematoxylin and eosin, kidney sections were also stained with the Ziehl-Neelsen method. Graphite furnace atomic absorption spectrophotometry was performed on all kidney samples. The presence of inclusion bodies and karyocytomegaly was scored. Six adult rats out of 21 (28.6%) showed kariocytomegaly and intranuclear inclusion bodies of the renal proximal tubules, characteristic of lead toxicity. The nuclear inclusion bodies were always Ziehl-Neelsen positive. The six rats which showed the characteristic histological alterations ascribed to lead, had lead concentrations statistically higher than the three control rats (4 microg/g wet wt. versus 0.09 microg/g wet wt., P<0.001) and than the captured wild rats. Intranuclear inclusion bodies were found in rats with kidney lead concentrations greater than 0.6 microg/g wet weight. There were significant differences between the four areas of capture. The incidence of intranuclear inclusion bodies and karyocytomegaly in proximal tubular cells was higher in rats with high lead levels detected by spectrophotometry. The present results indicate a renal lead concentration of 0.6 microg/g wet weight as a threshold to detect histologic kidney changes due to lead absorption. Results demonstrate that the examination of adult free-living rats may be considered an appropriate method to obtain a rapid, sensitive, easy, cost effective estimate of lead contamination in urban areas. Animal models allow us to evaluate potential human health risk associated with environmental contamination. PMID- 11916040 TI - Speciation and origin of particulate copper in runoff water from a Mediterranean vineyard catchment. AB - Fungicide treatments have led to large copper contents of the topsoils of most vineyards. This paper examines the contamination of surface waters by copper in a Mediterranean wine-growing catchment. Its aims were to characterise the forms of copper associated with suspended matter during a heavy autumn storm event and to identify which soils contribute the most to the copper exports. A mixing model involving three reservoirs, corresponding to three soil-landscape units (plateau, terraces and footslope-depression system) and two tracers (reducible iron content and dolomite/calcite ratio) was used to estimate the contribution of each reservoir to erosion during a storm flow. The average copper concentration of the suspended matter was 245 mg kg(-1), of which 1% was exchangeable, 4% acid soluble, 10% oxidizable, 23% reducible and 63% residual. The soils of the plateau of the catchment (chromic luvisols and haplic calcisols-FAO soil classification) were the source of 42% of copper exports but represented only 27% of the total catchment area. PMID- 11916041 TI - Investigation of unrecognized former secondary lead smelting sites: confirmation by historical sources and elemental ratios in soil. AB - Twelve suspected former secondary lead smelting sites were investigated. Ten of the sites were confirmed to be former secondary lead smelters or lead works by historical fire insurance maps and contemporaneous metal industry trade directories. At eight sites sampled, the ratio Sb:Pb was closer to ratios from 10 known lead smelting sites than were the As:Pb and Cd:Pb ratios. Data from the 10 known lead smelting sites showed that the Sb:Pb ratio is most characteristic of secondary lead smelting sites. This is because the primary alloy smelted at such sites is antimonial lead. Lead contamination at the eight sites investigated here can be attributed At least in part to the former smelters because of the association between Sb and Pb. PMID- 11916042 TI - Preliminary background ozone concentrations in the mountain and coastal areas of Bulgaria. AB - Urban and non-urban rural ozone (O3) concentrations are high in Bulgaria and often exceed the European AOT40 ecosystem as well as the AOT60 human health standards. This paper presents preliminary estimates to establish background, non urban O3 concentrations for the southern region of Bulgaria. Ozone concentrations from three distinctly different sites are presented: a mountain site influenced by mountain-valley wind flow; a coastal site influenced by sea-breeze wind flow; and a 1700-m mountain peak site without 'local' wind flow characteristics. The latter offers the best estimate of 46-50 ppb for a background O3 level. The highest non-urban hourly value, 118 ppb, was measured at the mountain-valley site. PMID- 11916043 TI - Sulphur migration in the soil-plant system contaminated by deposits from nickel industry: a field manipulation. AB - Sulphur migration in the soil-plant system was studied in a field experiment carried out in the spruce-pine forest 70 km SE from the 'Severonickel' smelting plant on the Kola Peninsula. In the experiment, dry particle deposits collected from filters of smokestacks in different departments of 'Severonickel' were applied on the study plots and on gravity lysimeters in quantities simulating estimated actual annual emissions in the area within 2-5 km from the smelter. In the present paper, the effects of two types of deposits consisting mostly of water-soluble sulphates of copper and nickel are described ("Cu-contaminant": Cu 55%, Ni-3.8%, SO4-25%; "Ni-contaminant": Ni-20%, Cu-5%, SO4--65%). Soil, plant tissue and penetration water were analysed during 3 years after the treatment. Extractable sulphur distribution in the soil-plant system showed downward movement after 1 year and upward movement after 3 years. The trend was more pronounced in the site treated with Ni-contaminant. PMID- 11916044 TI - Toxic effects of endocrine disrupters on freshwater sponges: common developmental abnormalities. AB - Endocrine disrupters are of substantial concern, in large part because effects of these compounds on the growth and development of many aquatic organisms are unknown. We examined toxic effects of three substances (ethylbenzene, nonylphenol, and bisphenol A), that are known to be hormonally active in many animals, on growth and development of two species of freshwater sponge. A common developmental abnormality was observed when sponges were treated with each of these compounds. The three compounds also caused significant reductions in growth rates. Lower concentrations resulted in malformed water vascular systems in several replicates. The utility of freshwater sponge bioassays is discussed as it relates to understanding possible mechanisms of action of endocrine disrupters on aquatic invertebrates. PMID- 11916045 TI - Kinetic study and modeling on photocatalytic degradation of para-chlorobenzoate at different light intensities. AB - In order to clarify the dependence of apparent adsorption constant Ks in the Langmuir-Hinshelwood (L-H) model on the incident light intensity, photocatalytic degradation kinetic characteristics were experimentally investigated at different light intensities using para-chlorobenzoate (4-CBA) as a model compound. In all cases 4-CBA initial degradation rates showed dependence on 4-CBA initial concentration, which can be described by the L-H model. However, the adsorption constant Ks and rate constant kr obviously varied with light intensity. To account for the experimental finding, slight modification of the classic kinetic model developed by Turchi and Ollis was tentatively proposed by assuming insufficiency of H2O/OH- available for photo-activated holes' scavenging. Such a kinetic model predicts that both k(r) and Ks(-1) are linearly proportional to the square root of the intensity in a rather large intensity range. The validity of the modified model was proved by fitting it to the experimental data. PMID- 11916046 TI - Extractable soil heavy metals following the cessation of biosolids application to agricultural soil. AB - Changes in soil heavy metal extractability following the cessation of biosolids applications were studied in a long-term field experiment. Two anaerobically digested biosolids from wastewater treatment plants in Madrid (Sur and Viveros) were applied to cropland from 1983 to 1990. Soil samples were collected in the 1st, 5th and 9th year after the last biosolids application. Soil pH did not vary significantly after biosolids applications. Organic matter and total heavy metals (Zn, Pb, Cd, Ni, Cr and Cu) concentrations initially increased but then declined over time, mostly after the first 5 years following biosolids application. Metal extracted with DPTA increased in Sur treatments during the 1st year and diminished thereafter. However, in Viveros treatments, heavy metals extracted increased during the 1st year, declined in 1995, and showed a slight increase in 1999. These changes in heavy metal extractability were widely observed in the percentage of extractable metal recovery (EMR). The differences observed in the pattern of the two sources of biosolids applied could be due to the different rates of decomposition of their organic matter. PMID- 11916047 TI - Plasma ALP activity and blood PCV value changes in chick fetuses due to exposure of the egg to different xenobiotics. AB - The effects of two polycyclic aromatic hydrocarbon (PAH) compounds (benz(k)fluoranthene and fluoranthene) and two heavy metals (cadmium and lead) were studied on the alkaline phosphatase (ALP) enzyme activity and hematocrit (PCV) values in chick fetuses. Eggs were exposed to one of the four compounds before incubation, by applying two exposure methods: injection (into the air cell) or immersion. Fluoranthene, cadmium and lead injection significantly decreased ALP activity compared to the control, while none of the compounds caused significant reduction in ALP activity in the immersion groups; however, a decreasing tendency was seen in these groups as well. Hematocrit values were increased after benz(k)fluoranthene immersion, fluoranthene immersion and cadmium injection. However, the doses in this study are relatively low (compared to other ecotoxicological studies in birds), the alteration in ALP enzyme activity and PCV values was apparent in each treatment group and is indicative of the high sensitivity of chick fetuses. PMID- 11916048 TI - Tolerance and metabolism of phenol and chloroderivatives by hairy root cultures of Daucus carota L. AB - Hairy root cultures are shown to be suitable experimental systems to screen higher plants for tolerance to various inorganic and organic pollutants, and for determining the role of the root matrix in the uptake and further metabolism of contaminants. A number of clones were obtained by infection of carrot tissues with Agrobacterium rhizogenes and two (the fastest and the slowest growing root clones) were chosen for further experimentation. Both clones showed a similar degree of tolerance towards phenol and its chlorinated derivatives, i.e. the growth of root biomass was maintained in concentrations of phenol equivalent to 1000 micromol/l, whilst the chlorophenols were tolerated only at concentrations 20 times lower (50 micromol/l). Transformed carrot roots were able to remove more than 90% of the exogenous phenolic compounds from the culture medium within 120 h after treatment. Metabolism of these compounds occurred in the root tissue and was accompanied by an increase in peroxidase activity. PMID- 11916049 TI - The spatial variability of nitrogen and phosphorus concentration in a sand aquifer influenced by onsite sewage treatment and disposal systems: a case study on St. George Island, Florida. AB - Groundwater from a shallow freshwater lens on St. George Island, a barrier island located in the Panhandle of Florida, eventually discharges into Apalachicola Bay or the Gulf of Mexico. Nutrient concentrations in groundwaters were monitored downfield from three onsite sewage treatment and disposal systems (OSTDS) on the island. Estimates of natural groundwater nutrient concentrations were obtained from an adjacent uninhabited island. Silicate, which was significantly higher in the imported drinking water relative to the surficial aquifer on St. George Island (12.2+/-1.9 mg Si l(-1) and 2.9+/-0.2 mg Si l(-1), respectively), was used as a natural conservative tracer. Our observations showed that nitrogen concentrations were attenuated to a greater extent than that of phosphorus relative to the conservative tracer. At the current setback distance (23 m), both nitrogen and phosphate concentrations are still elevated above natural levels by as much as 2 and 7 times, respectively. Increasing the setback distance to 50 m and raising the drainfields 1 m above the ground surface could reduce nutrient levels to natural concentrations (1.1+/-0.1 mg N l(-1), 0.20+/-0.02 mg P l(-1)). PMID- 11916050 TI - Consumption of biogenic nitric oxide in hydrated soil. AB - An experimental study was conducted in order to determine the relationship of nitric oxide (NO) consumption to water-filled pore space in soil. A test system that included the capability to blend gases, test soil samples, and analyze off gases was used to conduct the study. The experimental set consisted of three replicates at five different levels of soil water content and three different levels of soil nitrogen in a sandy loam soil: unamended soil, soil fertilized at 56.2 kg N per ha (50 lb N acre(-1)), and soil fertilized at 112.3 kg N per ha (100 lb N acre(-1)). The average NO consumption rates were 7.1x10(-13) g-NO cm( 3) soil, 3.5x10(-11) g-NO cm(-3) soil, and 1.5x10(-10) g-NO cm(-3) soil, respectively. PMID- 11916051 TI - Determination and analysis of sorption of 137Cs to soils in Hong Kong reservoir. AB - Soils from a local reservoir had been analyzed for their physical and chemical properties, distribution coefficients Kd and sorption rates for 137Cs. It was found that soils in Hong Kong were relatively low in organic matter and more acidic than those in some other countries. In sorption experiments, the rates of decrease in water activity concentration were initially very high, higher for larger mass of soils added, and then slowed down. The larger the mass, the shorter was the time for equilibrium. Three isotherms had been obtained by varying one of the three parameters (initial water activity concentration, slurry ratio and particle size) at a time. For constant initial water activity concentration and random sizes, percent sorption increased with slurry ratio. For soils of constant mass and random sizes. percent sorption decreased with increasing initial water activity concentration. For constant initial water activity concentration and slurry ratio, percent sorption increased with decreasing size of soil particles. A simple logistic model was developed to explain some of the findings. PMID- 11916052 TI - Biological and ecophysiological reactions of white wall rocket (Diplotaxis erucoides L.) grown on sewage sludge compost. AB - We studied the effects of sewage sludge compost on white wall rocket (Diplotaxis erucoides L.) compared with mineral fertilization and control (without any fertilizer) in a greenhouse experiment. The plants grown on the compost-amended soil showed a different growth dynamic: a significant delay in flowering and a bigger root system. Both the compost and the fertilization treatments increased biomass and seed yield. Heavy metal (Cu, Cd, Zn, Ni) distribution within the plant was in the following order: roots > leaves > stems, except for zinc which was homogeneously distributed. The balance of mineral nutrition was not affected by treatments. Zinc was the trace element which was most taken up. Unlike many species of Brassicaceae, white wall rocket is not a hyperaccumulator. Although sewage sludge compost improved plant growth, delay in flowering shows that it is necessary to take precautions when spreading sewage sludge in natural areas. PMID- 11916053 TI - What is your diagnosis? Malignant nerve sheath tumour. PMID- 11916054 TI - Association between anal furunculosis and colitis in the dog: preliminary observations. AB - Treatment of anal furunculosis in dogs is often unsatisfactory and may be associated with significant recurrence and complications. This may be compounded by the simultaneous presence of colitis in affected animals. Clinical signs associated with colitis and anal furunculosis may be similar, including faecal tenesmus, dyschezia and haematochezia. To examine the incidence of concurrent anal furunculosis and colitis, colonic biopsies were collected from 18 dogs referred for treatment of anal furunculosis. Nine dogs (50 per cent) had a histopathological diagnosis of colitis. Clinical signs more indicative of colitis than anal furunculosis (increased frequency of defecation, mucus in faeces and diarrhoea) were not observed more frequently in dogs with confirmed colitis compared with those with furunculosis alone. Therefore, while an association between colitis and anal furunculosis may exist, clinical signs alone cannot be used as an indicator of the presence of colitis in cases of anal furunculosis. The authors recommend that colonic biopsies should be undertaken in all dogs presented with anal furunculosis. Whether specific treatment of colitis in dogs with histopathological evidence of colitis improves the outcome of treatment for anal furunculosis awaits further study. PMID- 11916055 TI - Stereotactic CT-guided brain biopsy in the dog. AB - This study evaluated the accuracy of a new stereotactic CT-guided brain biopsy (SCTGBB) device on 23 client-owned dogs which presented with a brain lesion. Biopsy of the lesion was achieved in 95 per cent of cases. The target tissue was not sampled in one dog. Complications were observed in six dogs. Two dogs with highly vascularised brainstem tumours died after SCTGBB. Minor complications (slight variation in the neurological status) were observed in a further four cases. A diagnosis was reached in 16 dogs after cytological examination and in 21 dogs after histological evaluation. SCTGBB is an accurate diagnostic method for the diagnosis of brain lesions. PMID- 11916056 TI - Syncope associated with paroxysmal atrioventricular block an ventricular standstill in a cat. AB - A four-and-a-half-year-old neutered male shorthair cat was presented with a three week history of episodic fainting. Twenty-four hour electrocardiographic (Holter) recording revealed frequent prolonged episodes of complete atrioventricular (AV) block with ventricular standstill. The fainting episodes coincided with the longest periods of ventricular inactivity. A permanent transjugular pacemaker was placed to prevent further life-threatening episodes of syncope. Three months after discharge, the patient was re-presented with sudden onset dyspnoea associated with chylothorax and electrocardiography showed a third degree (complete) AV block. The pacemaker was reprogrammed in view of the third degree AV block and the chylous effusion was successfully treated by repeated thoracocentesis and long-term diuresis. However, the patient's general condition deteriorated progressively and the cat was euthanased five and a half months after implantation of the pacemaker. PMID- 11916057 TI - Pyogranulomatous skin disease and cellulitis in a cat caused by Rhodococcus equi. AB - This report describes a case of Rhodococcus equi infection causing pyogranulomatous skin disease and cellulitis in a two-year-old female domestic shorthaired cat. The case differed from previously reported cases in cats in its clinical presentation and in the locations of the lesions, which were similar to those seen in horses. The presence of an intracellular organism was confirmed by cytology and on histopathology. The aetiological diagnosis was confirmed by routine biochemical tests specific for R. equi on a pure isolate obtained from a biopsy specimen. The report also reviews the literature of the documented feline cases and discusses the common pitfalls in the diagnosis of such infections. PMID- 11916058 TI - Uterine carcinoma in a 10-month-old golden retriever. AB - A 10-month-old golden retriever was presented for investigation of reduced appetite, occasional vomiting and general dullness. Abdominal radiography demonstrated a large mid-abdominal soft tissue mass. Exploratory laparotomy identified a uterine mass, which was removed surgically. Histopathology confirmed uterine adenocarcinoma. Adjunctive chemotherapy with epirubicin was performed. The dog remained clinically normal without evidence of metastatic disease 24 months after surgery. Canine uterine adenocarcinoma is an extremely rare tumour and, as far as the authors are aware, this is the youngest reported case to date. PMID- 11916061 TI - Final steps to becoming a BSAVA standard practice. PMID- 11916059 TI - Retrieval of a patent ductus arteriosus coil following embolisation to the right subclavian artery. AB - A two-month-old female Tibetan terrier was re-presented with an incomplete closure of the patent ductus arteriosus. Following a second attempt to close the shunt by coil embolisation, arterial embolisation occurred. The successful removal of the embolised coil is described. PMID- 11916062 TI - New tool for the management of canine diabetes? PMID- 11916063 TI - Radionuclide and radiation protection data handbook 2nd edition (2002). AB - This handbook is a reference source of radionuclide and radiation protection information. Its purpose is to provide users of radionuclides in medicine, research and industry with consolidated and appropriate information and data to handle and transport radioactive substances safely. It is mainly intended for users in low and intermediate activity laboratories. Individual data sheets are provided for a wide range of commonly used radionuclides (144 in total). These radionuclides are classified into five different groups as a function of risk level, represented by colours red, orange, yellow, green and blue, in descending order of risk. PMID- 11916064 TI - Platelets: a new target for gene therapy of haemophilia B. PMID- 11916065 TI - P-selectin and PSGL-1: exploiting connections between inflammation and venous thrombosis. PMID- 11916066 TI - Expression of coagulation factor IX in a haematopoietic cell line. AB - We have developed a gene therapy project for haemophilia B which aims to express factor IX (FIX) in haematopoietic lineage. Haematopoietic stem cells and subsequent megakaryocyte-derived cells represent the target cells of this approach. Our speculation is that platelets can deliver the coagulation factor at the site of injury, and subsequently correct the haemostasis defect. In order to direct FIX expression in cells from the megakaryocytic lineage, we designed a FIX cassette where the FIX cDNA was placed under the control of the tissue-specific glycoprotein IIb (GPIIb) promoter. In stably transfected HEL cells, FIX production was higher when driven by the GPIIb promoter compared to the CMV promoter. Using a cassette containing both the GPIIb promoter and a truncated FIX intron 1, FIX synthesis was dramatically increased in HEL cells. Northern blot analysis demonstrated an increase in FIX mRNA amounts, which paralleled with an increase of FIX antigen in the culture supernatants. Using a one-stage clotting assay and an activation by FXIa and FVIIa/TF, the HEL-derived recombinant FIX was shown to be a biologically active protein. This recombinant protein exhibited a 60-kDa molecular mass and was more heterogeneous than plasma immunopurified FIX (Mononine). The molecular mass difference could be partly explained by a different glycosylation pattern. The GPIIb promoter appears therefore to be a very attractive sequence to specifically direct FIX production in the megakaryocytic compartment of hematopoietic cells. These data also demonstrate that hematopoietic cells may represent potential target cells in an approach to gene therapy of haemophilia B. PMID- 11916067 TI - New and effective treatment of experimentally induced venous thrombosis with anti inflammatory rPSGL-Ig. AB - BACKGROUND: P-selectin antagonism decreases thrombosis and inflammation in animal models of venous thrombosis (VT) prophylaxis. This study defines results using a P-selectin receptor antagonist for VT treatment. METHODS: Eight juvenile baboons underwent 6 h of iliofemoral venous stasis to produce an occlusive VT. Two days later, animals were treated for 14 days with rPSGL-Ig, 4 mg/kg (n3), LMWH (n2) or saline (n3) and treatment continued weekly (rPSGL-Ig) or daily (LMWH, saline). The animals were examined and sacrificed 14 days after treatment initiation (n4) or on day 90 (n4). RESULTS: Percent spontaneous vein reopening revealed a significant increase (p <0.05) in the proximal iliac vein in rPSGL-Ig and LMWH animals compared to controls (62%, 70% vs 8%), without differences in inflammation. No anticoagulation, thrombocytopenia, or wound complications were found in rPSGL-Ig animals. At 90 days, recanalization with iliac vein valve competence was found in treated animals. CONCLUSIONS: rPSGL-Ig successfully treated established VT without anticoagulation. PMID- 11916068 TI - Radionuclide synovectomy for hemophilic arthropathy: a comprehensive review of safety and efficacy and recommendation for a standardized treatment protocol. PMID- 11916069 TI - The mechanism of action of angiostatin: can you teach an old dog new tricks? AB - What is angiostatin? In 1994, Folkman and colleagues published a landmark paper describing anti-tumor effects in mice with a purified fragment of plasminogen they named angiostatin (1). Although many papers have been published describing activities of cryptic polypeptides derived from plasminogen fragments, this was the first report which associated plasminogen kringles 1-4 as a suppressor of metastasis development. This review will describe what is known about the mechanism of action of angiostatin from the current literature. PMID- 11916070 TI - Relationship of coagulation test abnormalities to tumour burden and postoperative DVT in resected colorectal cancer. AB - In a prospective study, coagulation test results were compared in 137 patients with colorectal cancer (CRC) and 39 subjects with benign colorectal diseases. Prothrombin fragment 1+2 (F1+2), thrombin-antithrombin complex (TAT), and soluble fibrin (SF) were measured in plasma before and after surgery. CRC patients presented with significantly higher values of F1+2 and TAT than controls. Patients with localised CRC had elevated values of F1+2 and TAT, whereas patients with advanced CRC also had elevated SF values. TAT and SF levels correlated with tumour spread, and normal values virtually excluded advanced cancer. Postoperative deep venous thrombosis (DVT) was diagnosed by phlebography in 20% of the CRC patients. Preoperative values of the markers did not predict postoperative DVT, but postoperative values did. PMID- 11916071 TI - Factor XIII-circulating levels and the Val34Leu polymorphism in the healthy male relatives of patients with severe coronary artery disease. AB - We aimed to investigate whether increased levels of FXIII and/or a low prevalence of the protective Leu allele (of the Val34Leu FXIII polymorphism) occur in relatives of patients with severe CAD. 185 healthy male relatives aged 65 or less were recruited from 125 patients with multi-vessel CAD and compared to 185 healthy, age-matched controls. The relatives and controls were similar in terms of clinical parameters. FXIII B-subunit levels were elevated in relatives, 1.11 microg/mL (1.08-1.14), compared with controls, 1.00 microg/mL (0.97-1.04), P <0.0001 but FXIII A2B2 levels did not differ between the groups. There was a strong correlation between FXIII B-subunit and the insulin resistance syndrome, however, adjusted B-subunit levels remained significantly higher in relatives. There was no difference in genotype frequency at the FXIII Val34Leu polymorphism between relatives and controls. FXIII B-subunit levels are elevated in the relatives of CAD patients and this is independent of other cardiovascular risk factors. PMID- 11916072 TI - Does inflammatory proteolytic activity contribute to the increased factor IX activation peptide in men at high risk of coronary heart disease? A preliminary study. AB - In the Second Northwick Park Heart Study, the activation peptides of factor IX (FIXpep) and factor X (FXpep) were measured in 1261 middle-aged men by double antibody radioimmunoassay. During follow-up 147 men who had a first coronary heart disease (CHD) event were found to have had an increased FIXpep (p = 0.003) and a reduced FXpep (p = 0.05) at baseline compared with those remaining CHD-free (controls). Plasma FIXpep and FXpep were positively associated, but the rate of rise in FIXpep with increasing FXpep was higher in cases than controls (p for interaction = 0.01). In a sample of 87 controls, FIXpep was positively and independently related to the concentrations of a polymorphonuclear-specific fibrinogen degradation product (p = 0.036) and FXpep (p = 0.004), but in larger samples no statistically significant associations were found either with C reactive protein or with fibrinogen concentration. The findings suggested that the increased FIXpep in men at high CHD-risk may have been partly due to the generation of factor IX inactivation peptides by inflammatory proteolysis and their recognition together with true FIXpep in the radioimmunoassay. Direct evidence for this hypothesis requires development of assays for human elastase specific factor IX inactivation peptides. PMID- 11916073 TI - Fibrinogen, viscosity and white blood cell count predict myocardial, but not cerebral infarction: evidence from the Caerphilly and Speedwell cohort. AB - Fibrinogen, plasma viscocity, and the white blood cell count predict ischaemic heart disease, but there is less certainty for their predictive power for ischaemic stroke. Studying stroke and ischaemic heart disease in the same cohort prospectively allows comparison of predictive strengths. The Caerphilly and Speedwell cohorts consist of a population sample of 4,860 men aged 45-59 years at recruitment who had baseline measurements of fibrinogen, plasma viscosity, and white blood cell counts. After 15-19 years of follow-up, men in the two cohorts experienced 312 ischaemic strokes and 557 ischaemic heart disease events. Mean fibrinogen, plasma viscosity and white blood cell counts differed significantly after adjustment for confounding factors between men with and without ischaemic heart disease, 0.25 g/l (95% CIs 0.1 8-0.32); 0.036 cp (95% CIs 0.027-0.044); 0.67 x 10(9)/l (95% CIs 0.50-0.84) respectively. The same measurements showed no significant differences after adjustment for the same confounding factors for men with and without ischaemic stroke, 0.05 g/l (95% CIs -0.04-0.14); 0.008 cp (95% CIs -0.003-0.019); 0.16 x 10(9)/l (95% CIs -0.06-0.38) respectively. PMID- 11916074 TI - Antibodies to factor XII are distinct from antibodies to prothrombin in patients with the anti-phospholipid syndrome. AB - Patients with the anti-phospholipid syndrome (APS) have antiphospholipid antibodies (aPA) which are often targeted towards phospholipid binding proteins such as beta2-glycoprotein I and prothrombin. Antibodies to factor XII (FXIIabs) have also been identified in some patients with APS. Factor XII (FXII) is a member of the kringle family of proteins which include plasminogen and prothrombin. Antibodies to prothrombin have been associated with myocardial infarction and have been shown to cross react with plasminogen. Sixteen patients with APS and FXIIabs were investigated for the presence of antibodies to prothrombin, by enzyme linked immunosorbent assay in a calcium (Ca++) independent assay. All sixteen showed different antibody binding patterns than those observed for antibodies to FXII. Eight patients were further investigated using surface plasmon resonance (SPR) for antibody binding to covalently bound FXII and to covalently bound prothrombin in both Ca++ dependent and independent systems. Of three patients demonstrating antibody binding to FXII by SPR, none demonstrated antibody binding to prothrombin in a Ca++ independent system with one demonstrating antibody binding to prothrombin that was Ca++ dependent. Of five patients who did not bind FXII by SPR, one demonstrated antibody binding to prothrombin in a Ca++ independent system while two demonstrated antibody binding to prothrombin in a Ca++ dependent system. Antibodies to FXII in patients with APS appear to be distinct from antibodies to prothrombin. PMID- 11916075 TI - Recombinant factor IX recovery and inhibitor safety: a Canadian post-licensure surveillance study. AB - As part of the Canadian post-licensure surveillance on the safety of recombinant factor IX (rFIX. BeneFIX), factor IX recovery and inhibitor development were studied. The recovery following rFIX infusion in 126 patients (mean = 0.77, median 0.72, range 0.36-1.85, 95% CI of mean 0.74-0.81, expressed as FIX activity increase in U/dL per IU FIX concentrate/kg body weight infused) was significantly lower than that following the last plasma-derived factor IX (pdFIX) infusion in 74 patients (mean 1.05, median 1.00, range 0.37-2.29, 95% CI of mean 0.99-0.97). The recovery for rFIX for patients aged < or =15 years (n = 41, mean recovery 0.64) and that for patients aged >15 years (n = 85, mean 0.84) was each significantly lower than that for pdFIX (aged < or =15 years: n = 21, mean recovery 0.91; aged >15 years: n = 53, mean recovery 1.10). For both rFIX and pdFIX concentrates, the recovery was lower in patients < or =15 years of age compared to those > 15 years of age. Similar data and conclusions were obtained on 66 patients with paired recovery data from rFIX and pdFIX. Overall, our data are similar to those obtained in formal clinical trials. Two of 244 patients treated with rFIX for up to 5 years have developed de novo inhibitors associated with anaphylaxis, an incidence that is similar to that reported for pdFIX. No other serious adverse events, including thrombotic episodes, were reported. To the best of our knowledge, this is the first formal report of recovery and inhibitor formation on rFIX in a peer-reviewed manuscript form. PMID- 11916076 TI - The utility of activated partial thromboplastin time (aPTT) clot waveform analysis in the investigation of hemophilia A patients with very low levels of factor VIII activity (FVIII:C). AB - The lower detection limit of the conventional one-stage aPTT based clotting assay for determining FVIII:C levels is generally 1.0-2.0 IU/dl. Consequently, it has been impossible to study the clinical significance of levels of FVIII:C less than 1.0 IU/dl. Using a photo-optical automated coagulation analyzer, the Organon Teknika MDA II, we have performed qualitative and quantitative aPTT waveform analysis and measured FVIII:C levels by automated one-stage aPTT clotting assay in 36 severely affected Hemophilia A patients. Qualitative waveform analysis showed clear evidence of individual differences in the waveform profile suggesting differing coagulant activity from patient to patient. The FVIII:C level was less than 0.2 IU/dl in 23 cases and levels of FVIII:C between 0.2 and 1.0 IU/dl could be discriminated in 13 patients. The FVIII:C level in these patients was closely correlated with the minimum value of the second derivative of the aPTT waveform (Min2). This is a measure of the acceleration of change in optical transmission at the initiation of coagulation. Furthermore, the correlation of the PMID- 11916077 TI - Procoagulant activity of T lymphoblastoid cells due to exposure of negatively charged phospholipid. AB - We have characterised the Procoagulant activity (PCA) of six well-established cell lines by assays of tissue factor (TF), thrombin and FXa generation, flow cytometry using Annexin V, and by binding studies with Factors VIII and IXa. The monocytic (THP-1 & U937) and promyelocytic (NB4) cells expressed high concentrations of TF antigen and activity, whereas TF in the lymphocytic cells (Molt 4, Jurkat & Nalm 6) was very low or absent. However the T-lymphoblastoid cells (Molt 4 & Jurkat) promoted the generation of large amounts of thrombin despite their low TF content, and these cells were also the most active in supporting Factor Xa generation. Molt 4 cells bound Factors VIII and IXa with high capacity and their activity was inhibited by Annexin V. These results indicate that the PCA of T-lymphoblastoid lines is due to expression of negatively charged phospholipids. Flow cytometry studies showed Annexin V binding to the major population of nonapoptotic Molt 4 cells and the PCA of Molt 4 was not increased when apoptosis was induced by staurosporine, indicating that PCA is independent of apoptotic status. PMID- 11916078 TI - Enhancing the anticoagulant potency of soluble tissue factor mutants by increasing their affinity to factor VIIa. AB - Complexation of factor VIIa (FVIIa) and tissue factor (TF) initiates the extrinsic pathway of blood coagulation. Inappropriate triggering of this pathway has been linked to thrombotic disorders. We have previously shown that a mutant form ofsoluble tissue factor (sTF, residues 1-219) having Lys165 and Lys166 replaced with alanine residues (hTFAA) gave an antithromboticeffect in a rabbit model of arterial thrombosis. Here we have improved the potency of hTFAA by incorporating amino acid substitutions thatincrease the affinity of sTF for FVIIa. Phage display has been used toselect consensus sequences at two FVIIa contact regions on sTF, segments 44-50 and 130-140, that individually showed modestly increased (approximately 2-fold) FVIIa-affinity. These consensus sequences have been combined with other previously selected mutations to generate a series of variants (hTFAA-2, hTFAA-3, and hTFAA-4) having 4 to 10-fold increased FVIIa-binding affinity. Surface plasmon resonance measurementsindicate that the increased affinity results mostly from an increase in the association rate constant. All of these variants displayed increased inhibitor potency relative to hTFAA in an assay of Factor X activation catalyzed by the complex of FVIIa with membrane TF. In addition, these affinity improved hTFAA variants are more potent anticoagulants and have increased antithrombotic activity relative to hTFAA in an ex vivo flow chamber model of thrombosis. The potency trend parallels increases in FVIIa-binding affinity; however, the absolute fold increases in potencies are greater than increases in binding affinity, consistent with kinetic studies of the FVIIa-binding event. Through incorporation of 10 amino acid substitutions (hTFAA-3) we have increased the antithrombotic activity of hTFAA by 20-fold. PMID- 11916079 TI - Human factor VIII inhibitor alloantibodies with a C2 epitope inhibit factor Xa catalyzed factor VIII activation: a new anti-factor VIII inhibitory mechanism. AB - Factor VIII (FVIII) inhibitor alloantibodies react with the A2, C2, or A3-CI domains of FVIII and inactivate FVIII activity. We recently demonstrated that an anti-C2 monoclonal antibody with a Val2248-Gly2285 epitope, inhibited factor Xa (FXa)-catalyzed FVIII activation, and that a FXa binding site for FVIII was located within residues Thr2253-Gln2270. In this study, we investigated whether anti-C2 alloantibodies inhibit FXa-catalyzed FVIII activation. Anti-C2 alloantibodies from four patients inhibited FVIII activation by FXa in one-stage clotting assay. Furthermore, analysis by SDS-PAGE showed that all alloantibodies inhibited FVIII proteolytic cleavage by FXa independently of phospholipid. To confirm direct inhibition of FVIII and FXa interaction, we examined the effect of alloantibodies on FVIII binding to anhydro-FXa, a catalytically inactive FXa, in ELISA. All alloantibodies and C2-affinity purified F(ab)'2 preparations inhibited FVIII binding to anhydro-FXa dose-dependently. Our results revealed a new inhibitory mechanism of FVIII, mediated by inhibition of FXa in the presence of anti-C2 alloantibodies. PMID- 11916080 TI - A comparative multi-laboratory assessment of three factor VIII/von Willebrand factor concentrates. AB - Five expert laboratories have participated in a cross-laboratory study to co evaluate and compare three commercial Factor VIII/von Willebrand factor (VWF) concentrates. A total of nine factor concentrate lots were evaluated, comprising AHF (High Purity) (AHF HP; x3), Biostate (x3) and Humate/Haemate (x3). All laboratories blind tested for FVIII: C, VWF: Ag and VWF: CB, four tested for VWF: RCo, and one performed VWF: Multimers. The study yielded inter-laboratory CVs for VWF: Ag and FVIII:C around 10-15%, and for VWF:CB and VWF:RCo around 20%, significantly lower than those of previous multi-laboratory surveys. All three lots of AHF HP contained in the vicinity of 25 U/ml FVIII:C, around 60-75 U/ml of VWF:Ag, but only 30-45 U/ml of VWF:CB and 40-50 U/ml of VWF:RCo (thus, CB/Ag ratio around 0.5-0.6 and RCo/Ag ratio around 0.6-0.7). Study determined that FVIII: C and VWF: RCo levels were similar to manufacturer assigned levels. Some loss of the high molecular weight (HMW) multimers was observed, together with an intense low molecular weight (LMW) VWF band consistent with some reduction or proteolysis of HMW VWF. All three lots of Humate/Haemate contained in the vicinity of 23-32 U/ml of FVIII:C, 70-105 U/ml of VWF: Ag, 50-90 U/ml of VWF: CB and VWF: RCo (i.e. CB/Ag ratio around 0.6-0.9 and RCo/Ag ratio around 0.6-1.1). Study-determined FVIII: C and VWF: RCo levels were similar to manufacturer assigned levels. The LMW multimer band seen with AHF HP was also observed with Humate/Haemate. All three lots of Biostate contained in the vicinity of 40-55 U/ml of FVIII:C, 105-170 U/ml of VWF:Ag, 90-150 U/ml of VWF:CB, and 90-135 U/ml of VWF:RCo (i.e. CB/Ag and RCo/Ag ratios around 0.7-1.0). Study-determined FVIII:C levels were similar to manufacturer-assigned levels. The LMW multimer band seen with AHF HP was not observed with Biostate. The defined pattern of increasing CB/ Ag from AHF HP to Humate/Haemate and Biostate was consistently observed in study data from each of the five laboratories. In conclusion, study findings indicate some differences in the retention of functional/ HMW VWF between factor concentrates, and this is expected to have significant implications in terms of clinical efficacy for therapy in VWD. PMID- 11916081 TI - Factor VII activation, apolipoprotein A-I and reverse cholesterol transport: possible relevance for postprandial lipaemia. AB - Postprandial lipaemia is associated with activation of factor VII (FVII) and efflux of cholesterol from tissues to nascent plasma high density lipoproteins (HDL) containing apolipoprotein A-I (apo A-I). To determine whether FVII activation and cholesterol efflux occur together in other situations, the responses to intravenous infusion of HDL-like apo A-I/phosphatidylcholine discs were measured in 10 healthy men. Disc infusion (40 mg apo A-I/kg body weight) over 4 h was followed by increases in HDL cholesteryl ester and plasma apo A-I (p <0.0001). Significant activation of FVII was apparent during infusion in fasting subjects (p = 0.03), activated FVII averaging 123% of baseline value by 12 h (p <0.0001). Plasma thrombin-antithrombin (TAT) complex increased to 156% of baseline level by 12 h (p >0.05) but individual responses differed considerably. Peak TAT post-infusion was associated inversely with peak HDL triglyceride concentration (p = 0.004). The coagulation responses to disc-infusion may be due to transfer of phosphatidylserine to cell surfaces during cholesterol efflux. PMID- 11916082 TI - Plasminogen has a broad extrahepatic distribution. AB - Plasmin is the major enzyme that dissolves fibrin in the vasculature and the predominant source of its zymogen, plasminogen, is liver. However, plasmin has a broad substrate spectrum and, if present in other tissues, may perform additional functions. We tested the hypothesis that plasminogen is expressed broadly extrahepatically. A sensitive and specific isotopic quantitative RT-PCR assay was developed to detect plasminogen mRNA from total RNA isolated from C57BL/6J mice tissues. Plasminogen mRNA was detected in adrenal, kidney, brain, testis, heart, lung, uterus, spleen, thymus and gut. Of these tissues, adrenal had the highest plasminogen mRNA content. In situ hybridization was utilized to localize plasminogen mRNA expressing cell types. Besides hepatocytes, positive cells were identified in both adrenal and kidney medullae and cortexes. Plasminogen mRNA expression was detected in cerebral, hippocampal and cerebellar neurons. Plasminogen mRNA was detected in cells in early stages of spermatogenesis in testis, present in the cortex and medulla of the thymus and in splenic white and red pulps. Our results suggest that the plasminogen gene is expressed broadly in extrahepatic tissues. Thus, tissues separated by local anatomic barriers as well as tissues accessible to circulating plasminogen have the capacity to provide local sources of plasminogen. PMID- 11916083 TI - Immunoglobulin G from patients with antiphospholipid syndrome impairs the fibrin dissolution with plasmin. AB - Immunoglobulin G (IgG) isolated from normal human blood plasma stabilizes the structure of perfused crosslinked fibrin and prolongs the time for its dissolution with plasmin, when the fibrin surface is exposed to 500 s(-1) shear rate flow. The IgG from patients suffering in antiphospholipid syndrome with thrombotic complications exerts even stronger antifibrinolytic effect. A patient, whose IgG does not affect the fibrin dissolution with plasmin, displays a bleeding tendency. The shear stress-induced disassembly of the fibrin clots containing IgGs with antifibrinolytic potency occurs at a much more advanced stage of fibrin digestion, as evidenced by the electrophoretic pattern of the ureatreated samples. The antifibrinolytic effects are also produced under static conditions and these are caused by the variable portion of the IgG molecules (fragment Fab), whereas the constant part (fragment Fc) has no inhibitory effect. The IgGs with antifibrinolytic properties do not affect directly the plasmin activity in amidolytic assay, but the IgGs from APS patients obliterate the competition of the fibrin and the peptidyl-p-nitroanilide for the protease in the same assay system suggesting interference of the IgGs with the plasmin action on the fibrin substrate. Thus, the correlation of the clinical symptoms with the effect of the isolated IgG on the dissolution of perfused fibrin clots supports a physiological and a pathological role of IgG in the fibrinolytic process related to the variability of the cross-reactions of immunoglobulins with fibrin, fibrin degradation products or fibrin-plasmin complexes. PMID- 11916084 TI - Proline-rich tyrosine kinase 2 and focal adhesion kinase are involved in different phases of platelet activation by vWF. AB - Stimulation of human platelets with von Willebrand factor (vWF) induces the rapid tyrosine phosphorylation of several proteins, but very little is known on the tyrosine kinases involved in this process. In the present work, we investigated and compared the activation of two related tyrosine kinases expressed in platelets: the proline-rich tyrosine kinase 2 (Pyk2) and the focal adhesion kinase (FAK). Both kinases were tyrosine phosphorylated upon vWF interaction with glycoprotein Ib-IX-V complex, but with different mechanisms. Tyrosine phosphorylation of FAK was totally dependent on thromboxane A2 production, and was inhibited by the integrin alphaIIbeta3 antagonist RGDS peptide. Moreover, chelation of intracellular calcium or inhibition of protein kinase C (PKC) totally blocked vWF-induced tyrosine phosphorylation of FAK, indicating that this event is downstream phospholipase A2 and phospholipase C activation. By contrast, tyrosine phosphorylation of Pyk2 was only partially reduced by aspirin and RGDS, and was not affected by either calcium chelation or PKC inhibition, suggesting that activation of this kinase does not require phospholipase-mediated signalling. Both FAK and Pyk2 translocated to the cytoskeleton upon vWF stimulation of human platelets by a mechanism depending on agonist-induced actin polymerisation. Prevention of cytoskeletal relocation of Pyk2 and FAK by cytochalasin D totally blocked vWF-induced tyrosine phosphorylation of both kinases. Finally, phosphorylation of Pyk2 induced by vWF, but not by thrombin, was inhibited by piceatannol, suggesting that this kinase lies downstream Syk. These results demonstrate that both Pyk2 and FAK are involved in platelet stimulation by vWF, but indicate that only Pyk2 may play a role in the early signal transduction events activated by ligand binding to glycoprotein Ib-IX-V. PMID- 11916085 TI - Hydroxychloroquine reverses platelet activation induced by human IgG antiphospholipid antibodies. AB - Prothrombotic properties of antiphospholipid (aPL) antibodies may be explained in part by their ability to enhance the activation of platelets pre-treated with low doses of ADP or thrombin. The antimalarial drug hydroxychloroquine (HQ) has been used successfully in prevention of postoperative thrombosis and in treatment of patients with SLE or APS. In one study, administration of HQ reversed the thrombogenic properties of aPL in mice. However, the mechanism of action of HQ in preventing thrombosis is not clearly understood. In order to explore this further, the effects of HQ on activation of platelets by aPL in the presence of a thrombin agonist was studied. The changes in the expression of GPIIb/IIIa (CD41a) and GPIIIa (CD61) on platelet membrane by flow cytometry were used as indicators of platelet activation. Citrated whole blood from a healthy donor was treated at room temperature with suboptimal doses of a thrombin agonist receptor peptide (TRAP) and affinity-purified aPL antibodies, in the presence and in the absence of hydroxychloroquine (1 mM). TRAP increased the expression of GPIIb/IIIa and GPIIIa on platelet surface. The treatment of the platelets with the six aPL antibodies in the presence of 12 nMol/ml TRAP further increased the expression of GPIIb/IIIa by 42.3+/-12.3% and the expression of GPIIIa was further incremented by 46.8+/-13.5%. The effects of aPL and TRAP on expression of platelet surface markers of activation was completely abrogated by HQ in a dose-dependent fashion and was effective at concentrations of HQ as low as 25 microg/ml (0.0125 mM). This suggests at least one possible mechanism by which HQ may prevent thrombosis. This may have important implications in treatment of thrombosis in APS patients. PMID- 11916086 TI - 15 deoxy delta12,14 PGJ2 induces procoagulant activity in cultured human endothelial cells. AB - 15 deoxy delta12,14 PGJ2 (15d-PGJ2), a high affinity ligand of peroxisome proliferator-activated receptor gamma (PPARgamma) has been proposed to act as a negative feedback regulator of the inflammatory response. We investigated the effect of 15d-PGJ2 on the anticoagulant property of endothelial cells. 15d-PGJ2 stimulated a moderate but sustained increase in tissue factor (TF) activity in HUVECs and EA.hy926 cells while causing a partial loss of thrombomodulin (TM) activity. When cells were co-treated with 15d-PGJ2 and TNF-alpha, the subsequent elevation of TF activity was synergistically increased over that of cells treated with TNF-alpha alone and the decline of TF activity after 24 h was less marked than TNF-alpha alone. The induction of TF by 15d-PGJ2 alone and in combination with TNF-alpha was reduced in the presence of PD 98059, suggesting the participation of the MEK/ERK pathway. The thiazolidinedione PPARgamma agonist ciglitazone had no effect on TF levels but reduced the expression of endothelial protein C receptor. The ability of 15d-PGJ2 to enhance a procoagulant phenotype arising from TNF-alpha suggests a pro-inflammatory role for the prostaglandin. PMID- 11916087 TI - Adipocyte hypertrophy in stromelysin-3 deficient mice with nutritionally induced obesity. AB - Several matrix metalloproteinases (MMPs), including the stromelysins MMP-3 and MMP-11, are expressed in adipose tissue. To investigate a potential role of MMP 11 (stromelysin-3) in adipose tissue development, five-week-old male wild-type mice (MMP-11+/+) or mice with deficiency of MMP-11 (MMP-11-/-) were fed a high fat diet (HFD, 42% fat) for 15 weeks. Haematologic parameters, including white and red blood cells, platelets, haemoglobin and haematocrit, and metabolic parameters including glucose, triglycerides and total cholesterol were not different for both genotypes. At the time of sacrifice, the body weight of the MMP-11-/- mice was higher than that of the MMP-11+/+ mice (36+/-1.4 g versus 29+/ 0.9 g, p = 0.0002). The weight of the isolated subcutaneous (SC) and gonadal (GON) fat deposits was also higher in MMP-11-/- mice (620+/-150 mg versus 280+/ 28 mg for SC fat, and 970+/-180 mg versus 430+/-62 mg, p < 0.05, for GON fat). Adipocytes in MMP-11-/- adipose tissue were hypertrophic as compared to MMP-11+/+ adipocytes (volume of 57+/-12 x 10(3) microm3 versus 31+/-2.4 x 10(3) microm3 for SC fat, and 100+/-18 x 10(3) microm3 versus 57+/-7.6 x 10(3) microm3 for GON fat; both p < 0.06). In nutritionally induced obesity models in mice a potential role of the fibrinolytic system was suggested in adipocyte hypertrophy. The hypertrophy observed in this model is, however, not related to changes in fibrinolytic parameters, as suggested by our finding that levels of t-PA, u-PA and PAI-1 antigen as well as t-PA and u-PA activity were not different in SC or GON adipose tissue extracts of both genotypes. As the main biological function of MMP-11 remains unknown, it is not clear whether the adipocyte hypertrophy in MMP 11-/- adipose tissue is directly related to the deficiency or to other pathways affected by MMP-11. PMID- 11916088 TI - Enoxaparin sodium improves pregnancy outcome in aspirin-resistant antiphospholipid/antiprotein antibody syndromes. PMID- 11916089 TI - Enoxaparin and unfractionated heparin in acute coronary syndromes without ST segment elevation. PMID- 11916090 TI - Catecholamines, via beta-adrenoceptors, increase intracellular concentrations of 3',5'-cyclic guanosine monophosphate (cGMP) through nitric oxide in human platelets. PMID- 11916091 TI - The Arg200Trp mutation in the human tissue factor gene. PMID- 11916092 TI - Successful surgical hemostasis in patients with von Willebrand disease following infusion of KoateDVI. PMID- 11916093 TI - Detection of transfused platelets in a patient with Glanzmann thrombasthenia. PMID- 11916094 TI - The differences in thrombolytic effects of administrated recombinant t-PA between Japanese and Caucasians. PMID- 11916095 TI - Method comparison of flow cytometric assay of platelet microparticles and changes of platelet microparticles during cancer chemotherapy. PMID- 11916096 TI - Rebuttal to: von Willebrand factor-cleaving protease in childhood diarrhoea associated haemolytic uraemic syndrome. PMID- 11916097 TI - Thrombin generation in a reconstituted system: a comment. PMID- 11916099 TI - Sibutramine in clinical practice. AB - The role of sibutramine as an effective adjunct to diet and exercise in weight management programmes has been proven in clinical trials. Although the reality of managing obesity in clinical practice is very different from that of a controlled clinical trial, there is good evidence that sibutramine is an easy-to-use and highly effective additional therapy when used in everyday practice to manage obesity and its complications. It is possible to predict which patients will respond well to sibutramine treatment, and to modify programmes and support to ensure the best outcome for patients, whatever their clinical circumstances. PMID- 11916100 TI - Developing an action plan for obesity education. AB - Obesity education has long been neglected. Medical training continues to devote little time to education on obesity and its comorbidities, and practising physicians often feel ill equipped and poorly supported when dealing with overweight and obese patients. A European action plan for obesity education is required, to act at all levels--government, education providers and trainers, media, health professionals and patients-to improve understanding of obesity as a chronic but manageable medical condition. PMID- 11916101 TI - When, for whom and how to use sibutramine? AB - Sibutramine is the first of a new generation of weight management drugs, and offers long-term control of weight when used as an adjunct to diet and exercise. This paper considers the criteria set down by the European Union's regulatory agency for its use and how this relates to the information now available on its efficacy, side-effect profile and benefits in different patient groups. The data presented relate to 18-65y olds; its use in other age groups has not been established. The EU licence permits continuous treatment for periods of up to 1 y; the STORM study has now led the US Food and Drug Administration to extend clearance to 2 y. The addition of sibutramine to a regimen of diet, exercise and lifestyle modification results in 3-5 times more patients responding to a weight reduction programme, ie achieving more than 5% weight maintenance. Sibutramine is indicated for use in obese patients (BMI > 30 kg/m2) and also overweight patients with a BMI > 27 kg/m2 who have additional obesity-related risk factors such as type 2 diabetes or dyslipidaemia. PMID- 11916102 TI - Sibutramine in overweight/obese hypertensive patients. AB - Obesity and hypertension are closely associated risk factors for cardiovascular disease. An increase in body mass index is commonly associated with an increase in blood pressure and weight reduction is recommended as the principal intervention in the obese hypertensive patient. Sibutramine therapy, as part of a programme of diet and exercise, can help achieve marked weight reduction and improve metabolic and other cardiovascular risk factors. In hypertensive patients, weight loss induced by sibutramine has also been found to reduce blood pressure. Studies have shown that in the obese, well-controlled hypertensive patient, sibutramine is a safe and well-tolerated therapy offering the many clinical benefits associated with weight reduction in this high-risk population. PMID- 11916103 TI - The importance of obesity in diabetes and its treatment with sibutramine. AB - Weight gain is a known risk factor for the development of type 2 diabetes and even modest weight reduction can reduce the risk of developing diabetes, so controlling body weight is an important public health goal in the fight against diabetes and its comorbidities. Weight reduction is also a cornerstone of diabetes management, improving glycaemic control and reducing other risk factors associated with this disease. Pharmacotherapies such as sibutramine contribute to the management of type 2 diabetes in overweight and obese patients. PMID- 11916104 TI - Obesity through the ages of man. AB - From before birth and onwards there is the potential for obesity. Many of the predisposing factors can be prevented or minimised through educational and preventative strategies that support healthier diet and lifestyles, but the need for active treatment and management of obesity has never been greater. Childhood and adolescence are crucial times for prevention, and treatment in middle age confers important health and quality of life benefits. As obesity becomes more prevalent and the population ages, new and inventive strategies are required to tackle this ubiquitous health problem also in older age groups. PMID- 11916105 TI - The role of sibutramine in weight management--towards a blueprint for a sibutramine weight management system. AB - Sibutramine offers a new opportunity to provide a comprehensive weight reduction programme for many overweight and obese patients. A methodical and team approach to managing patients on sibutramine plus diet and exercise should hold the key to the best use of this adjunctive therapy in real clinical practice. This paper provides an overview of how to care for patients who are in need of weight management, and shows that simple measures and well organised, good clinical practice are all that is required to achieve clinically important weight reduction and longer term care. PMID- 11916106 TI - How does sibutramine work? AB - Sibutramine offers three types of benefit in weight management: by enhancing weight loss, by improving weight maintenance and by reducing the comorbidities of obesity. The clinical effects of sibutramine are explained through its known mode of action as a serotonin (5-HT) and noradrenaline reuptake inhibitor (SNRI). This dual mechanism of action results in two synergistic physiological effects--a reduction in energy intake and an increase in energy expenditure, which combine to promote and maintain weight loss. PMID- 11916107 TI - Modulation of calcium balance in tilapia larvae (Oreochromis mossambicus) acclimated to low-calcium environments. AB - This study examined how developing fish larvae regulate their Ca2+ balance for acclimation to low ambient Ca2+. Calcium balance in newly hatched larvae was examined individually. Developing larvae not only increased Ca2+ influx but also decreased Ca2+ efflux when they were acclimated to low-Ca2+ environments. After acclimation for 8 days, the influx and efflux of the low-Ca2+ (0.02 mM) group were about 106% and 43%, respectively, compared to those of the high-Ca2+ (1.0 mM) group. Sensitivity and response to low-Ca2+ environments are age-dependent. Upon acute exposure to low Ca2+. newly hatched (H0) larvae increased both Ca2+ influx (from 24% to 67% of high-Ca2+) and net uptake (from 5% to 69%) within 64 h, while 3-day-posthatching (H3) larvae managed to reach the levels of the control within 38 h. Declining Ca2+ efflux in H3 larvae occurred 14 h after exposure, much faster than those in H0 larvae (38 h). It is suggested that modulation of Ca2+-balance mechanisms in developing larvae is dependent upon the levels of Ca2+ in the larval body. PMID- 11916108 TI - Oxygen consumption and force development in turtle and trout cardiac muscle during acidosis and high extracellular potassium. AB - Relative to species such as rainbow trout, freshwater turtle shows a high tolerance to challenges involving acidosis and increases in extracellular K+. Therefore, the effects of acidosis or high K+ on twitch force and oxygen consumption were examined in ventricular ring preparations from these two species. The oxygen consumption associated with force development was estimated by net oxygen consumption (oxygen consumption during twitch force development minus that during rest). For turtle, elevation of CO2 from 2% (pH 7.7) to 12% (pH 6.9) in the gas equilibrating the muscle bath decreased twitch force by 20% without any effects on oxygen consumption. Decreasing pH from 7.7 to 6.9 with 22 mM lactic acid had similar effects. For trout, CO2-induced acidosis decreased twitch force by approximately 60%. Furthermore, force development became energetically less efficient as it fell disproportionately more than net oxygen consumption. This was not observed for lactic acidosis. For trout but not for turtle, acidosis resulted in an increase in oxygen consumption during rest. An increase in extracellular K+ from 2.5 mM to 10 mM depressed force and oxygen consumption proportionately for both species. Adrenaline (10 microM) increased twitch force for both species and oxygen consumption for trout; it attenuated the effects of high extracellular K+. Neither adrenaline nor high K+ influenced the ratio of force to net oxygen consumption. As opposed to high extracellular K+, acidosis appears to increase the energetic cost of contractility, particularly for the trout heart. PMID- 11916109 TI - Ionoregulatory development and the effect of chronic silver exposure on growth, survival, and sublethal indicators of toxicity in early life stages of rainbow trout (Oncorhynchus mykiss). AB - Rainbow trout embryos and larvae were continuously exposed, in a flow-through system, to 0, 0.1 microg/l (measured=0.098 +/- 0.002 microg/l) or 1.0 microg/l (measured=0.853+/-0.022 microg/l) total silver (as AgNO3) in moderately hard water (120 mg CaCO3/l, 0.70 mM Cl, 1.3 mg/l dissolved organic matter and 13.7 +/- 0.1 degrees C) from fertilization to I week post-hatch. The objectives of the study were to investigate the effects of chronic silver exposure on mortality, time to hatch and growth, and on sublethal physiological indicators of toxicity. Exposure to 1.0 microg/l total silver resulted in a small, but statistically significant, increase in mortality (16%) relative to controls (12%) but interestingly, resulted in an increased rate of growth (as indicated by larval weight, length and extractable protein) and ionoregulatory development over the duration of this study. Whole body unidirectional Na uptake (J(in)Na+) increased with silver exposure concentration (both 0.1 microg/l and 1.0 microg/l total silver) just prior to and following hatch, with up to a three-fold elevation in J(in)Na+ in the 1.0 microg/l treatment relative to controls. Qualitatively similar changes in whole body Na+,K-ATPase activity (per mg protein or per whole embryo or larvae) also occurred over this period. By 1 week post-hatch, there were no differences in J(in)Na among treatments and Na+,K+-ATPase activity levels in silver exposed groups were significantly reduced relative to controls. Within 2 days following hatch, there was an elevation in whole larval ammonia levels, while cortisol levels were elevated at 1 week post-hatch in the 1.0 microg/l treatment relative to controls. Ionoregulatory disturbance and elevations in both cortisol and ammonia have also been observed during acute silver exposure in adult rainbow trout, indicating that chronic and acute mechanisms of toxicity may be similar. PMID- 11916110 TI - The inhibition of ice nucleators by insect antifreeze proteins is enhanced by glycerol and citrate. AB - Antifreeze proteins depress the freezing point of water while not affecting the melting point, producing a characteristic difference in freezing and melting points termed thermal hysteresis. Larvae of the beetle Dendroides canadensis accumulate potent antifreeze proteins (DAFPs) in their hemolymph and gut, but to achieve high levels of thermal hysteresis requires enhancers, such as glycerol. DAFPs have previously been shown to inhibit the activity of bacterial and hemolymph protein ice nucleators, however, the effect was not large and therefore the effectiveness of the DAFPs in promoting supercooling of the larvae in winter was doubtful. However, this study demonstrates that DAFPs, in combination with the thermal hysteresis enhancers glycerol (1 M) or citrate (0.5 M), eliminated the activity of hemolymph protein ice nucleators and Pseudomonas syringae ice nucleating active bacteria, and lowered the supercooling points (nucleation temperatures) of aqueous solutions containing these ice nucleators to those of water or buffer alone. This shows that the DAFPs, along with glycerol, play a critical role in promoting hemolymph supercooling in overwintering D. canadensis. Also, DAFPs in combination with enhancers may be useful in applications which require inhibition of ice nucleators. PMID- 11916111 TI - The role of the symbiotic fungus in the digestive metabolism of two species of fungus-growing ants. AB - Leaf-cutting ants live in an obligatory symbiosis with a fungus which they grow on fresh leaves harvested by workers. This study attempts to clarify the respective role of ants and fungus in the degradation of plant material, in order to highlight the evolutionary basis of this mutualistic association. The symbiotic system of two ant species, Acromyrmex subterraneus subterraneus and Acromyrmex crassispinus, was investigated. To identify the digestive carbohydrases, a total of 19 specific and synthetic plant material substrates were tested on workers from different castes (major and minor), larvae and fungus. Extracts of A. subterraneus and A. crassispinus workers showed high enzymatic activity particularly on starch, maltose, sucrose and alpha-1,4 glucoside. Larvae degraded starch, sucrose, maltose but also laminarin, and all the detected activities were higher than those found for workers. The symbiotic fungus of A. subterraneus was mostly active on laminarin, xylan and cellulose, while the symbiotic fungus of A. crassispinus was mostly active on laminarin, starch, maltose and sucrose. The enzymatic activities of ants and fungus belonging to the same symbiotic system tended not to overlap, suggesting that the association is highly evolved and of an ancient origin. PMID- 11916112 TI - Effect of aestivation on muscle characteristics and locomotor performance in the green-striped burrowing frog, Cyclorana alboguttata. AB - The Green-striped burrowing frog, Cyclorana alboguttata survives extended drought periods by burrowing underground and aestivating. These frogs remain immobile within cocoons of shed skin and mucus during aestivation and emerge from their burrows upon heavy rains to feed and reproduce. Extended periods of immobilisation in mammals typically result in muscle atrophy and a decrease in muscle performance. We examined the effect of aestivation and hence prolonged immobilisation, on skeletal muscle mass, in vitro muscle performance, and locomotor performance in C. alboguttata. Frogs were aestivated in soil for 3 months and were compared with control animals that remained active, were fed, and had a continual supply of water. Compared to the controls, the wet mass of the gastrocnemius, sartorius, gracilus major, semimembranosus, peroneus, extensor cruris, tibialis posticus and tibialis anticus longus of aestivators remained unchanged indicating no muscle atrophy. The in-vitro performance characteristics of the gastrocnemius muscle were maintained and burst swimming speed was unaffected, requiring no recovery from the extended period of immobilisation associated with aestivation. This preservation of muscle size, contractile condition and locomotor performance through aestivation enables C. alboguttata to compress their life history into unpredictable windows of opportunity, whenever heavy rains occur. PMID- 11916113 TI - Postnatal development and thermoregulation in the precocial European hare (Lepus europaeus). AB - We investigated postnatal development of captive juvenile European hares (Lepus europaeus) with respect to growth, food intake and thermoregulation. Leverets increased their body weight on average 8.5-fold within 35 days, with a peak in relative growth rate on day 8. Although leverets started to take up solid food in the 2nd week of lactation, milk was the main energy source. Leverets of larger litters (two or three young) had less energy supply via milk than those of smaller litters (one young), and digested more energy in form of solid food. Weaning weight was positively correlated with body weight at birth and decreased with increasing litter size. Metabolic rates were largely determined by ambient temperature (Ta) and the increasing body weights. Leverets were able to maintain normothermic body temperatures from the Ist day of life during cold exposure down to Ta -8 degrees C. In their 1st week of life leverets showed reduced rates of heat loss in the cold, possibly by using peripheral vasoconstriction. Our results suggest that the precocial development of leverets is characterised by high maintenance costs due to rapid growth and thermoregulation, which may lead to negative energy balances when food supply is inadequate and T(a)s are low. We hypothesise that these physiological characteristics have played an important role in the recent decline of European hare populations. PMID- 11916114 TI - Origin and evolution of arthropod hemocyanins and related proteins. AB - Arthropod hemocyanins are large, multimeric, (n x 6) copper-containing proteins that deliver oxygen in the haemolymph of many chelicerate, crustacean, myriapod, and also possibly some insect species. The arthropod hemocyanins belong to a large protein superfamily that also includes the arthropod phenoloxidases, certain crustacean and insect storage proteins (pseudo-hemocyanins and hexamerins), and the insect hexamerin receptors. Here I summarise the present knowledge of the origin, functional adaptations, and evolution of these proteins. Arthropod and mollusc hemocyanins are, if at all, only distantly related. As early as in the arthropod stem line, the hemocyanins emerged from a phenoloxidase like enzyme. The evolution of distinct hemocyanin subunits, as well as the formation of multi-hexamers occurred independently within the arthropod subphyla. Hemocyanin subunit evolution is strikingly different in the Chelicerata, Myriapoda and Crustacea. Hemocyanins individually gave rise to two distinct copper-less storage proteins, the insect hexamerins and the crustacean pseudo hemocyanins (cryptocyanins). The receptor responsible for the uptake of hexamerin by the larval fat body of the insects emerged from a hexamerin-precursor. Molecular phylogenetic analyses show a close relationship of the crustacean and insect proteins, providing strong support for a pancrustacean taxon, while structural data suggest a myriapod-chelicerate clade. PMID- 11916115 TI - How hydrogeology can save the world. PMID- 11916116 TI - Using high hydraulic conductivity nodes to simulate seepage lakes. AB - In a typical ground water flow model, lakes are represented by specified head nodes requiring that lake levels be known a priori. To remove this limitation, previous researchers assigned high hydraulic conductivity (K) values to nodes that represent a lake, under the assumption that the simulated head at the nodes in the high-K zone accurately reflects lake level. The solution should also produce a constant water level across the lake. We developed a model of a simple hypothetical ground water/lake system to test whether solutions using high-K lake nodes are sensitive to the value of K selected to represent the lake. Results show that the larger the contrast between the K of the aquifer and the K of the lake nodes, the smaller the error tolerance required for the solution to converge. For our test problem, a contrast of three orders of magnitude produced a head difference across the lake of 0.005 m under a regional gradient of the order of 10(-3) m/m, while a contrast of four orders of magnitude produced a head difference of 0.001 m. The high-K method was then used to simulate lake levels in Pretty Lake, Wisconsin. Results for both the hypothetical system and the application to Pretty Lake compared favorably with results using a lake package developed for MODFLOW (Merritt and Konikow 2000). While our results demonstrate that the high-K method accurately simulates lake levels, this method has more cumbersome postprocessing and longer run times than the same problem simulated using the lake package. PMID- 11916117 TI - Ground water flow analysis of a mid-Atlantic outer coastal plain watershed, Virginia, U.S.A. AB - Models for ground water flow (MODFLOW) and particle tracking (MODPATH) were used to determine ground water flow patterns, principal ground water discharge and recharge zones, and estimates of ground water travel times in an unconfined ground water system of an outer coastal plain watershed on the Delmarva Peninsula, Virginia. By coupling recharge and discharge zones within the watershed, flowpath analysis can provide a method to locate and implement specific management strategies within a watershed to reduce ground water nitrogen loading to surface water. A monitoring well network was installed in Eyreville Creek watershed, a first-order creek, to determine hydraulic conductivities and spatial and temporal variations in hydraulic heads for use in model calibration. Ground water flow patterns indicated the convergence of flow along the four surface water features of the watershed; primary discharge areas were in the nontidal portions of the watershed. Ground water recharge zones corresponded to the surface water features with minimal development of a regional ground water system. Predicted ground water velocities varied between < 0.01 to 0.24 m/day, with elevated values associated with discharge areas and areas of convergence along surface water features. Some ground water residence times exceeded 100 years, although average residence times ranged between 16 and 21 years; approximately 95% of the ground water resource would reflect land use activities within the last 50 years. PMID- 11916118 TI - Improving a regional model using reduced complexity and parameter estimation. AB - The availability of powerful desktop computers and graphical user interfaces for ground water flow models makes possible the construction of ever more complex models. A proposed copper-zinc sulfide mine in northern Wisconsin offers a unique case in which the same hydrologic system has been modeled using a variety of techniques covering a wide range of sophistication and complexity. Early in the permitting process, simple numerical models were used to evaluate the necessary amount of water to be pumped from the mine, reductions in streamflow, and the drawdowns in the regional aquifer. More complex models have subsequently been used in an attempt to refine the predictions. Even after so much modeling effort, questions regarding the accuracy and reliability of the predictions remain. We have performed a new analysis of the proposed mine using the two-dimensional analytic element code GFLOW coupled with the nonlinear parameter estimation code UCODE. The new model is parsimonious, containing fewer than 10 parameters, and covers a region several times larger in areal extent than any of the previous models. The model demonstrates the suitability of analytic element codes for use with parameter estimation codes. The simplified model results are similar to the more complex models; predicted mine inflows and UCODE-derived 95% confidence intervals are consistent with the previous predictions. More important, the large areal extent of the model allowed us to examine hydrological features not included in the previous models, resulting in new insights about the effects that far-field boundary conditions can have on near-field model calibration and parameterization. In this case, the addition of surface water runoff into a lake in the headwaters of a stream while holding recharge constant moved a regional ground watershed divide and resulted in some of the added water being captured by the adjoining basin. Finally, a simple analytical solution was used to clarify the GFLOW model's prediction that, for a model that is properly calibrated for heads, regional drawdowns are relatively unaffected by the choice of aquifer properties, but that mine inflows are strongly affected. Paradoxically, by reducing model complexity, we have increased the understanding gained from the modeling effort. PMID- 11916119 TI - Estimate of recharge of a rising water table in semiarid Niger from 3H and 14C modeling. AB - A hydrodynamic survey carried out in semiarid southwest Niger revealed an increase in the unconfined ground water reserves of approximately 10% over the last 50 years due to the clearing of native vegetation. Isotopic samplings (3H, 18O, 2H for water and 14C, 13C for the dissolved inorganic carbon) were performed on about 3500 km2 of this silty aquifer to characterize recharge. Stable isotope analyses confirmed the indirect recharge process that had already been shown by hydrodynamic surveys and suggested the tracers are exclusively of atmospheric origin. An analytical model that takes into account the long-term rise in the water table was used to interpret 3H and 14C contents in ground water. The natural, preclearing median annual renewal rate (i.e., recharge as a fraction of the saturated aquifer volume) lies between 0.04% and 0.06%. For representative characteristics of the aquifer (30 m of saturated thickness, porosity between 10% and 25%), this implies a recharge of between 1 and 5 mm/year, which is much lower than the estimates of 20 to 50 mm/year for recent years, obtained using hydrological and hydrodynamic methods and the same aquifer parameters. Our study, therefore, reveals that land clearing in semiarid Niger increased ground water recharge by about one order of magnitude. PMID- 11916121 TI - Using direct current resistivity sounding and geostatistics to aid in hydrogeological studies in the Choshuichi alluvial fan, Taiwan. AB - Ground water reservoirs in the Choshuichi alluvial fan, central western Taiwan, were investigated using direct-current (DC) resistivity soundings at 190 locations, combined with hydrogeological measurements from 37 wells. In addition, attempts were made to calculate aquifer transmissivity from both surface DC resistivity measurements and geostatistically derived predictions of aquifer properties. DC resistivity sounding data are highly correlated to the hydraulic parameters in the Choshuichi alluvial fan. By estimating the spatial distribution of hydraulic conductivity from the kriged well data and the cokriged thickness of the correlative aquifer from both resistivity sounding data and well information, the transmissivity of the aquifer at each location can be obtained from the product of kriged hydraulic conductivity and computed thickness of the geoelectric layer. Thus, the spatial variation of the transmissivities in the study area is obtained. Our work is more comparable to Ahmed et al. (1988) than to the work of Niwas and Singhal (1981). The first "constraint" from Niwas and Singhal's work is a result of their use of linear regression. The geostatistical approach taken here (and by Ahmed et al. [1988]) is a natural improvement on the linear regression approach. PMID- 11916122 TI - Back diffusion of chlorinated solvent contaminants from a natural aquitard to a remediated aquifer under well-controlled field conditions: predictions and measurements. AB - Vertical profiles of tetrachloroethene (or perchloroethylene, PCE) and trichloroethene (TCE) were used to validate a diffusion process in a natural aquitard at Dover Air Force Base, Delaware. PCE and TCE distributions in the aquitard underlying an unconfined aquifer were sampled from core tubes obtained at four times over the course of a 35-month field investigation within "test cells" that were isolated from the surrounding ground water by means of grout sealed steel sheetpile barriers (Mackay et al. 2000). For the final 23 months of this period, boundary conditions at the aquifer/aquitard interface were such that a "back diffusion" of contaminants from the aquitard was induced. Modeling predictions of concentration changes were made on the basis of the earliest coring results and an assumption of sorption-retarded diffusion and using laboratory information about sorption and diffusion characteristics of the media. The predictive modeling was complicated by the fact that "initial" and "final" PCE and TCE distributions in the aquitard were measured at different (albeit proximate) coring locations, such that results reflect spatial variations in aquitard characteristics. This problem was solved by means of an inverse interpretation that involved spatial "translation" of observed profiles on the basis of the laboratory characterizations and assuming a common aquifer-side contaminant history. Predictions indicated substantial change in PCE and TCE concentrations within the upper aquitard (near the aquifer/aquitard interface) and the development of a back-diffusion profile up into the aquifer. Modeling also predicted comparatively minor profile changes in the deeper aquitard, and especially in the deep layer where sorption was strongest. All of these predicted effects were observed in the coring results. Although not exact, the agreement between predictions and observations was sufficiently good to justify the basic tenets of the diffusion model and to support a conclusion that major processes of advection and/or transformation were unimportant within the 35-month time scale of this work. PMID- 11916120 TI - More realistic soil cleanup standards with dual-equilibrium desorption. AB - The desorption of contaminants from soils/sediments is one of the most important processes controlling contaminant transport and environmental risks. None of the currently adopted desorption models can accurately quantify desorption at relatively low concentrations; these models often overestimate the desorption and thus the risks of hydrophobic organic chemicals, such as benzene and chlorinated solvents. In reality, desorption is generally found to be biphasic, with two soil phase compartments. A new dual-equilibrium desorption (DED) model has been developed to account for the biphasic desorption. This model has been tested using a wide range of laboratory and field data and has been used to explain key observations related to underground storage tank plumes. The DED model relates the amount of a chemical sorbed to the aqueous concentration, with simple parameters including octanol-water partition coefficient, solubility, and fractional organic carbon; thus, it is the only biphasic model, to date, that is based on readily available parameters. The DED model can be easily incorporated into standard risk and transport models. According to this model, many regulatory standards of soils and sediments could be increased without increasing the risks. PMID- 11916123 TI - Kriging water levels with a regional-linear and point-logarithmic drift. AB - Ground water levels measured in the vicinity of pumping wells are kriged using a regional-linear and point-logarithmic drift, the latter derived from the approximation to the Theis equation for drawdown in response to a pumping well. Kriging is widely used throughout the hydrogeologic discipline, most commonly as the preferred method for constructing gridded hydrogeologic datasets suitable for contouring. Residuals arising from using the most common (linear) drift to krige water levels in the vicinity of extraction wells often indicate large local departures from the linear drift, which correlate with areas of drawdown. The combined regional-linear and point-logarithmic drift accounts for these drawdowns using a logarithmic approximation for the curvature of the potentiometric surface. The drift model approximates the principal physical processes that govern ground water flow and ultimately govern the autocorrelation of ground water elevation data. This approach produces maps of contoured water levels that more realistically represent physical conditions and allow for improved interpretation of measured water-level data by including features and information known to be present. Additional benefits include an improved estimate of the regional (background) hydraulic gradient and generation of an approximately flow conserved grid suitable for two-dimensional particle tracking. PMID- 11916124 TI - Generating MODFLOW grids from boundary representation solid models. AB - Complex stratigraphy can be difficult to simulate in MODFLOW models. MODFLOW uses a structured grid that requires that each grid layer be continuous throughout the model domain. This makes it difficult to explicitly represent common features such as pinchouts and embedded seams in a MODFLOW model. In this paper, we describe a method for automatically generating MODFLOW-compatible grids from boundary-representation solid models. Solid models are data structures developed originally for computer-aided design applications that define the geometry of three-dimensional objects. Solid models can be used to represent arbitrarily complex stratigraphy. The elevations defined by the solids are then extracted from the solids in a manner that preserves the continuous-layer requirement imposed by MODFLOW. Two basic approaches are described: The first method adjusts the MODFLOW grid dimensions (layer elevations) to fit the solid model boundaries, and the second method creates a regular MODFLOW grid and adjusts the material properties to match the changes in stratigraphy. One of the main benefits of using solid models to define stratigraphy for MODFLOW models is that it provides a grid-independent definition of the layer elevations that can be used to immediately re-create the MODFLOW grid geometry after any change to the grid resolution. PMID- 11916125 TI - Nonparametric method for transmissivity distributions along boreholes. AB - The transmissivities of individual fractures along a borehole are difficult to obtain unless each fracture is tested. To estimate a fracture-transmissivity distribution from section transmissivities, a method was developed based on fixed interval-length transmissivities and the corresponding number of fractures for each interval. The method is nonparametric and iterative, and the fractures are viewed as two-dimensional features, in which the total transmissivity of a borehole is equal to the sum of individual fracture transmissivities. Initially, a linear a priori assumption of the transmissivity distribution is made, and from this a so-called mean transmissivity function is derived. Subsequently, the mean transmissivity of the Nj fractures within a section, j, of the borehole is estimated, and the same value of the mean transmissivity function represents Nj possible fracture transmissivities from the initial distribution. This is repeated for each borehole section, and, eventually, all fracture transmissivities are sorted to give the next iteration's transmissivity distribution and the corresponding mean transmissivity function. Finally, the distributions converge, yielding a possible fracture-transmissivity distribution. The method was verified for a synthetic data sample and then tested on a sample from a borehole at the Aspo Hard Rock Laboratory, Sweden. For the synthetic data, the method gave a distribution that was fairly close to the original one; for the Aspo data, 15% of the fractures had a transmissivity larger than the measurement limit (1 x 10(-9) m2/sec), and these transmissivities follow a log-normal distribution. PMID- 11916126 TI - The history of hydrology at Stanford University. PMID- 11916127 TI - Mutant laboratory mice with abnormalities in pigmentation: annotated tables. AB - Mammalian pigment cell research has recently entered a phase of significantly increased activity due largely to the exploitation of the many mutant mouse stocks that are coming on stream. Numerous transgenic, targeted mutagenesis (so called 'knockouts'), conditional (so-called 'gene switch') and spontaneous mutant mice develop abnormal coat color phenotypes. The number of mice that exhibit such abnormalities is increasing exponentially as genetic engineering methods become routine. Since defined abnormalities in such mutant mice provide important clues to the as yet often poorly understood functional roles of many gene products, this overview includes a corresponding, annotated table of mutant mice with pigmentation alterations. These range from early developmental defects via a large array of coat color abnormalities to a melanoma metastasis model. This overview should provide helpful pointers to investigators who are looking for mouse models to explore or to compare functional activities of genes of interest and for comparing coat color phenotypes of spontaneous or genetically engineered mouse mutants with novel ones. Secondly, this review includes a table of mouse models of specific human diseases with genetically defined pigmentation abnormalities. In summary, this annotated table should serve as a useful reference for anyone interested in the molecular controls of pigmentation. PMID- 11916129 TI - CD35 expression on peripheral blood granulocytes of patients with atopic dermatitis. AB - We examined CD35 expression on granulocytes from 45 patients with atopic dermatitis (AD) (male, 21, female, 24) and 25 age and sex-matched controls (male, 15; female, 10). There was no significant difference in the peripheral blood neutrophil count between AD patients and controls, whereas the eosinophil count in AD patients was significantly higher than that of controls. The percentages of CD35 positive eosinophils and neutrophils were determined by using two-color flow cytometric analysis. As regards eosinophils, we found CD35 expression from AD patients to be lower than that of controls (P < 0.05), but there was no correlation between the CD35 expression and disease severity. In contrast, the CD35 expression on neutrophils from AD patients was much lower than that of controls (P < 0.005). Furthermore, CD35 expression on neutrophils of severe AD group was significantly higher than that of the mild AD group (P < 0.05). This suggests that the CD35 expression of neutrophils but not eosinophils reflects disease severity in AD patients and the CD35 expression on neutrophils in AD patients may associate with susceptibility to bacterial infection. PMID- 11916128 TI - Histamine-induced IL-6 and IL-8 production are differentially modulated by IFN gamma and IL-4 in human keratinocytes. AB - It is known that large amounts of histamine are stored in mast cells located in the superficial dermis of the skin and can be released upon appropriate stimulation. However, the effects of histamine on keratinocyte function have not been well characterized. We therefore examined the capacity of histamine to modulate the production of interleukin (IL)-6 and IL-8 by keratinocytes. We found that histamine significantly augmented the production of IL-6 and IL-8 in a dose- and time-dependent manner. The enhancing effects of histamine were completely inhibited by a potent H1 receptor (H1R) antagonist, emedastine difumarate. Pyrilamine (a much weaker H1R antagonist) and cimetidine (an H2R antagonist) only partially inhibited the enhancing effects of histamine. The histamine-induced up regulation of IL-6 and IL-8 production, however, was completely abrogated by a combination of pyrilamine and cimetidine. The IL-6 production was significantly enhanced by interferon (IFN)-gamma. Interestingly, IFN-gamma and IL-4 both significantly augmented the histamine-induced IL-6 production. On the other hand, the production of IL-8 was inhibited by IFN-gamma, and IFN-gamma and IL-4 both completely abrogated the histamine-induced IL-8 production. These results suggest that the histamine-induced IL-6 production and IL-8 production are differentially regulated by IFN-gamma and IL-4. Histamine may be an important modulator of cytokine production in epidermal milieu. PMID- 11916131 TI - Expression of 36-kDa microfibril-associated glycoprotein (MAGP-36) in human keratinocytes and its localization in skin. AB - Microfibril-associated glycoprotein-36 (MAGP-36) is a recently isolated elastin binding protein and considered to be a member of microfibril-associated glycoproteins (MAGPs). We studied the expression of MAGP-36 in cultured normal human keratinocytes and its localization in the skin. MAGP-36 was found to be expressed in cultured human keratinocytes by Western blot and RT-PCR assays. The levels of MAGP-36 (polypeptide and mRNA) and the number of MAGP-36-producing keratinocytes were greatly increased during Ca(2+)-induced differentiation of keratinocytes. Immunohistochemical studies demonstrated that MAGP-36 colocalized with elastic fibers and formed candelabra like-fibers in the superficial dermis of normal skin. In the elderly skin of sun-exposed region, immunoreactivity of MAGP-36 in the superficial dermis disappeared. In the lesional skin of pseudoxanthoma elasticum which is an elastin-related disorder, immunoreactivity of MAGP-36 was found in the accumulation of disintegrated elastic fibers. The results show that MAGP-36 is a component of elastic fibers in the dermis and co operates with elastin in normal and diseased conditions. PMID- 11916130 TI - Cultured human dermal papilla cells secrete a chemotactic factor for melanocytes. AB - Large numbers of pigmented melanocytes are located in human hair follicles, predominantly around the dermal papillae, and the number of melanocytes and the melanogenic activity of the hair follicles are closely related to the hair cycle. We found that cultured human dermal papilla cells secreted a melanocyte chemoattractant into the medium. Skin fibroblasts also showed weak chemoattraction of melanocytes, while skin keratinocytes and melanocytes did not. Since this chemotactic activity was heat-and protease-sensitive and was present in the relatively high molecular weight fraction (130-200 kDa), it may be due to extracellular matrix (ECM) that proteins secreted from the cultured dermal papilla cells. This chemotactic signal between dermal papillae and melanocytes may control the localization and migration of hair melanocytes in vivo. PMID- 11916132 TI - Cicatricial pemphigoid differs from bullous pemphigoid and pemphigoid gestationis regarding the fine specificity of autoantibodies to the BP180 NC16A domain. AB - Bullous pemphigoid (BP), pemphigoid (herpes) gestationis (PG), cicatricial pemphigoid (CP), and lichen planus pemphigoides (LPP) are autoimmune subepidermal bullous diseases that are characterized by circulating autoantibodies to the transmembrane hemidesmosomal protein BP180/type XVII collagen. Previous studies demonstrated that the majority of patients with BP, PG, and LPP show antibodies to an immunodominant, membrane-proximal non-collagenous domain (NC16A) on the extracellular portion of BP180. By the use of non-overlapping peptides of the NC16A domain, we previously demonstrated that autoantibodies from BP and PG patients mainly react with epitopes clustered within the N-terminus of this immunodominant site of BP180; antibodies from patients with LPP also recognized the C-terminal portion of NC16A. However, some of these results had been obtained indirectly by preadsorption studies. The aim of the present study was to analyze the fine specificity of IgG autoantibodies to NC16A in sera from patients with CP and to compare their reactivity with antibodies from BP, PG, and LPP patients using a series of new overlapping fragments covering the entire NC16A domain. We confirm that BP and PG sera mainly react with N-terminal epitopes of NC16A, whereas sera from patients with LPP also bind to C-terminal portions, of this domain. Interestingly, out of ten patients with CP, the sera of seven reacted with NC16A; within NC16A, these sera bound to both C-terminal fragments and an N terminal epitope right next to the cell membrane. Our data demonstrate a heterogeneous binding pattern of autoantibodies to BP180 NC16A in patients with CP. PMID- 11916133 TI - The effect of antibiotics on the production of superantigen from Staphylococcus aureus isolated from atopic dermatitis. AB - Staphylococcus aureus (S. aureus) often colonizes on the skin of patients with atopic dermatitis. It is known that superantigens which are staphylococcal enterotoxins can activate T cells without processing by antigen presenting cells. It has been suggested that activated T cells release various cytokines which may exacerbate or prolong the cutaneous inflammation associated with atopic dermatitis. Reduction of bacterial colonization from skin lesions has been reported to be effective in the treatment of atopic dermatitis. Therefore, antimicrobial therapy using antibiotics may be a treatment option for atopic dermatitis in selected patients. We examined the effect of antibiotics on the production of superantigen from S. aureus in vitro to determine the action mechanism of antibiotics in the treatment of atopic dermatitis. It was found that antibiotics with inhibitory effect on protein synthesis can suppress the production of superantigen. On the other hand, the superantigen production was not suppressed by antibiotics having either the inhibitory effect on cell wall synthesis or on nucleic acid synthesis. Levels of the suppressive effect on superantigen production by S. aureus varied with strains tested in this study. Moreover, we demonstrated that replication of DNA coding of superantigen produced by S. aureus was suppressed only by roxithromycin (ROX), which is a new macrolide. This finding suggests that ROX may have an effect at the gene level. These results suggested that the suppressive effects of antimicrobial agents that act as inhibitors of protein synthesis on superantigen production from S. aureus may be useful in the treatment of atopic dermatitis. PMID- 11916134 TI - Is the photoprotective effect of vitamin E based on its antioxidative capacity? AB - Vitamin E (alpha-tocopherol) is widely used in the prevention of ultraviolet (UV) induced skin damages. Mostly, the photoprotective effect of vitamin E is attributed to its radical scavenger capacity. We discuss here other possible explanations for its preventive potential. PMID- 11916136 TI - Kinetics of inactivation of Ulva pertusa Kjellm alkaline phosphatase by ethylenediaminetetraacetic acid disodium. AB - Ulva pertusa Kjellm alkaline phosphatase (EC 3.3.3.1) is a metalloenzyme, the active site of which contains a tight cluster of two zinc ions and one magnesium ion. The kinetic theory described by Tsou of the substrate reaction during irreversible inhibition of enzyme activity has been employed to study the kinetics of the course of inactivation of the enzyme by EDTA. The kinetics of the substrate reaction at different concentrations of the substrate p-nitrophenyl phosphate (PNPP) and inactivator EDTA indicated a complexing mechanism for inactivation by, and substrate competition with, EDTA at the active site. The inactivation kinetics are single phasic, showing that the initial formation of an enzyme-EDTA complex is a relative rapid reaction, following by a slow inactivation step that probably involves a conformational change of the enzyme. The presence of Zn2+ apparently stabilizes an active-site conformation required for enzyme activity. PMID- 11916135 TI - Non-linear slow-binding inhibition of Aerococcus viridans lactate oxidase by Cibacron Blue 3GA. AB - Lactate oxidase (LOD) was purified from cells of Aerococcus viridans by phase partitioning in Triton X-114 (TX-114), ammonium sulphate fractionation and FPLC ion exchange chromatography. The purification achieved from a crude extract of A. viridans was 32-fold with a 60% recovery of activity. The isolated enzyme was a true FMN-containing LOD in tetrameric form with a subunit molecular weight of 48,000. The KM for L-lactate was 175 microM, a 6-fold less value than described in the literature. Among the inhibitors tested, Cibacron Blue 3GA showed the lowest Ki. At low concentrations, Cibacron Blue 3GA behaved as a dye-, pH- and time-dependent inhibitor. A Dixon plot of the steady-state rate showed the time dependent inhibition to be non-linear, contrary to that described for other slow binding inhibitors. A model to explain this phenomenon was proposed. The model implies the binding of Cibacron Blue 3GA to the isomerised form of the initial enzyme-inhibition complex (E'I). PMID- 11916137 TI - Quantitative structure-activity relationships (QSARs) within series of inhibitors for mammalian cytochromes P450 (CYPs). AB - The results of quantitative structure-activity relationship (QSAR) studies on series of P450 inhibitors are reported. Cytochrome P450 families CYP1, CYP2 and CYP51 have been investigated for QSAR analysis, including those of CYP2 subfamilies: CYP2A, CYP2B, CYP2C, CYP2D and CYP2E. The accumulated evidence indicates different structural descriptors being involved, depending on the P450 enzyme concerned, although compound lipophilicity in the form of either logP or logD(7.4) appears to represent a common factor in some cases. This is thought to represent desolvation of the P450 active site, although quadratic expressions in lipophilicity tend to suggest that membrane transport is important, especially for CYP2B and CYP2E isoforms. In general, there is close agreement (R = 0.95 0.99) between experimental pKi values and those calculated via QSAR analysis. PMID- 11916138 TI - Quantitative structure-activity relationship study of novel alpha1a-selective adrenoceptor antagonists. AB - Two series of compounds were recently reported as novel alpha1a-selective adrenoceptor antagonists. In the first series, a dihydropyrimidone moiety is attached to a 4-phenyl piperidine containing side chain, while in the second, it is linked to a 4-substituted phenyl piperazine containing side chain. These compounds having potential for the treatment of benign prostatic hyperplasia, a urological disorder in the older age male population, were subjected to a quantitative structure-activity relationship study. The analysis has helped to ascertain the role of different substituents in explaining the observed binding potencies of these analogues. In the first category of compounds, three sites R1, R2, and X were varied and from the quantitative structure-activity relationship, it emerged that X- and R1-substituents having respectively, the high values of field and resonance effects may lead to more potent alpha1a-antagonists. The substituent of R2, being either CH3 or C2H5, does not add to improve the activity and thus the site, at present, becomes redundant. This site may, however, be explored for some additional substituents in future. In the second series of compounds, the phenyl ring, linked to a piperazine moiety at the end of a side chain, was substituted with various groups onto different positions. From derived significant correlations, it appeared the less polar and/or bulky substituents at the meta- and para-positions and a more hydrophobic substituent at the para position are advantageous. PMID- 11916139 TI - A new class of potent reversible inhibitors of metallo-proteinases: C-terminal thiol-peptides as zinc-coordinating ligands. AB - A number of substrate analogous peptides containing a phosphoramidate, phosphonate ester, hydroxamate, carboxylate or sulfhydryl group are known to be inhibitors of thermolysin and other metalloproteinases. According to the specificity, most of the inhibitors mimic the prime site of the active center. Hitherto, peptidyl derivatives with a thiol group at the C-terminus have not been described. We have synthesized the protected cysteamides Ac-Ala-Ala-CA-SH and Z Aa1-Aa2-CA-SH (Aa1: Ala, Pro; Aa2: Ala, Leu). The binding of these thiol peptide inhibitors to the metalloproteinases is characterized first by the coordination of the thiolate group of the inhibitor to the catalytic zinc ion and second by the subsite interaction of the peptide ligand in the active site of the enzyme. All peptide derivatives were competitive inhibitors of the zinc metalloproteinase thermolysin. The strongest inhibition was found with Z-Pro-Leu-CA-SH (Ki = 30 microM). Substitution of the N-protecting benzyloxycarbonyl residue towards the acetyl group in the peptide inhibitor, the inhibition constant decreased about 25 times. PMID- 11916140 TI - Carbonic anhydrase activity modulators: synthesis of inhibitors and activators incorporating 2-substituted-thiazol-4-yl-methyl scaffolds. AB - A small series of 2-[4-(4-substituted-phenylsulfonyl)phenyl]-4 chloromethylthiazoles has been used as a scaffold for the preparation of carbonic anhydrase (CA) inhibitors and activators. For obtaining CA inhibitors, zinc binding functions of the sulfamide and sulfamate type have been introduced into the molecules of these compounds, by reaction of the chloromethyl derivatives with sodium sulfamide/sodium sulfamate. For obtaining CA activators, the primary amino function has been introduced in these molecules by means of the Gabriel syntheses. The new sulfamide/sulfamates were effective CA II and CA IV inhibitors, but showed no inhibitory activity against isozyme I. The new amines on the other hand were much more effective CA I, II and IV activators compared to histamine, the lead compound used for their synthesis. PMID- 11916142 TI - Probing the active site of pea seedlings amine oxidase with optical antipodes of sedamine alkaloids. AB - Interactions of pea seedlings amine oxidase (PSAO, EC 1.4.3.6) with sedamine derivatives were studied. All compounds exhibited a competitive inhibition with the inhibition constants in the range 0.03-1.0 mM. The inhibition effect increased in the order allosedamine < sedamine << norallosedamine < norsedamine. The nor-derivatives are about five-fold stronger inhibitors and the allo-isomers are slightly weaker inhibitors than the others. Interestingly, the (-) diastereomers of the studied sedamines were considerably stronger inhibitors than the (+)-antipodes. Absorption spectroscopy was used to differentiate between two known groups of competitive inhibitors of PSAO. A representative of substrate analogues, 1,5-diamino-3-pentanone, bleached the spectrum of the TPQ cofactor producing a very stable intermediate of the enzyme catalytic cycle that was only slowly converted to the product. On the other hand, the alkaloids did not perturb the spectrum of TPQ so they may interact with some other residue near the active site. PMID- 11916141 TI - Inhibition of glutamate carboxypeptidase II by phosphonamidothionate derivatives of glutamic acid. AB - A limited series of N-thiophosphonyl-glutamates were found to be inhibitors of the prostate-specific membrane antigen (PSMA) form of glutamate carboxypeptidase II. Comparative inhibitory profiles of an analogous O-thiophosphonyl-2 hydroxyglutarate revealed that the amido-linkage of the N-thiophosphonyl glutamate provides a significant enhancement of inhibitory potency presumably due to significant hydrogen-bonding interactions with acceptor groups in the active site of PSMA resulting in tighter binding. An analogous N-phosphonyl-glutamate exhibited significantly greater inhibitory potency than the parent N thiophosphonyl-glutamate indicating that the sulfur ligand of the N thiophosphonyl-glutamates is responsible for less favorable active-site interactions than oxygen, potentially due to steric crowding from the longer P-S bond or as a result of active-site metal substitution of Co(II) for Zn(II) arising from assay conditions. PMID- 11916143 TI - Isomerase activity of the C-terminal fructose-6-phosphate binding domain of glucosamine-6-phosphate synthase from Escherichia coli. AB - The isomerase activity of the C-terminal fructose-6P binding domain (residues 241 608) of glucosamine-6-phosphate synthase from Escherichia coli has been studied. The equilibrium constant of the C-terminal domain k(eq) ([glucose-6P]/[fructose-6 P]) = 5.0. A non-competitive product inhibition of the isomerase activity by the reaction product glucose-6-P has been detected. The existence of more than one binding and reaction sites for the substrate fructose-6P on the molecule of glucosamine-6-phosphate synthase can be expected. The fructose-6P binding domain possibly includes a regulatory site, different from the catalytic center of the enzyme. PMID- 11916144 TI - Inhibition of succinate-cytochrome C reductase by a ferromacrocyclic complex. AB - Succinate-cytochrome c reductase (SCR) from mouse liver was inhibited strongly and reversibly by an iron (II) macrocyclic complex 3. The inhibition was observed for the enzyme toward the reduction of both 2,6-dichlorophenol indophenol (DCIP) and cytochrome c (cyt c). The inhibition was a mixed type and noncompetitive with respect to the reduction of DCIP and cyt c, respectively. Values of the inhibition constant ranged from 6.6 to 8.3 microM. The IC50 for the complex 3 was found to be 16.6 +/- 0.8 and 12.1 +/- 0.5 microM for the enzyme toward DCIP and cyt c, respectively. The reduced form of complex 3 also exhibited enzyme inhibition but to a less extent. Complex 3, at a lower level, equal to 25% of its LD50 showed about 50% inhibition of the enzyme through in vivo dose-dependent effect. These findings suggested that the structure of the equatorial benzoquinoid macrocyclic ligand of the Fe(II) complex is involved in the enzyme inhibition. PMID- 11916145 TI - Progress curves analysis as an alternative for exploration of activation inhibition phenomena in cholinesterases. AB - The kinetic behaviour of insect acetylcholinesterases deviates from the Michaelis Menten pattern. These deviations are known as activation or inhibition at various substrate concentrations and can be more or less observable depending on mutations around the active site of the enzyme. Most kinetic studies on these enzymes still rely on initial rate measurements. It is demonstrated here that according to this method one of the deviations can be overlooked. We attempt to point out that in such cases a detailed step-by-step progress curves analysis is successful. The study is focused on two different methods of analysing progress curves: (i) the first one is based on an integrated initial rate equation which can sufficiently fit truncated progress curves under corresponding conditions; and (ii) the other one precludes the algebraic formulae, but uses numerical integration for searching a non analytical solution of ordinary differential equations describing a kinetic model. All methods are tested on three different acetylcholinesterase mutants from Drosophila melanogaster. The results indicate that kinetic parameters for the E107K mutant with highly expressive activation and inhibition can be well evaluated applying any analysis method. It is quite different for E107W and E107Y mutants where latent activation is present, but discovered only using one or the other progress curves analysis methods. PMID- 11916146 TI - Enantioselectivity of some 1-[(benzofuran-2-yl) phenylmethyl] imidazoles as aromatase (P450AROM) inhibitors. AB - The enantioselectivity ratio ((+)-:(-)-forms) of three substituted 1-[(benzofuran 2-yl) phenylmethyl] imidazoles as inhibitors of aromatase (P450AROM) was 2.16, 12.3 and 1.0 for the 4-methyl-, 4-fluoro- and 4-chloro-substituted compounds, respectively. The (+/-)-compounds were all >1000 times more potent than (+/-) aminoglutethimide (IC50 = 12 x 10(3) nM). High potency (5.3-65.0 nM) for all the enantiomers studied is unusual since activity usually resides in one form for chiral inhibitors of P450AROM. The 4-methyl derivative was fitted into the model [Furet, P., Batzl, C., Bhatnager, A.S., Francotte, E., Rihs, G. and Lang, M. (1993) J. Med. Chem. 36, pp. 1393-1400] for binding of S-(-)-fadrazole to the active site and the (R)- and (S)- forms both gave a good fitting pattern with (S) (-)-fadrazole so accounting for their close activity. Docking of both forms into the active site model for P450AROM [Laughton, C.A., Zvelebil, M.J.J.M. and Neidel, S. (1993) J. Steroid Biochem. Mol. Biol. 44, pp. 399-407], using the orientation of (S)-(-)-fadrazole, gave similar strong binding along the position of the C and D rings of the steroid substrate and in the hydrophobic cavity below the A/B rings. The site was probed for group size accommodation using the less potent 4-phenyl analogue (IC50(+/-) = 242 nM): the (S)-form showed restricted access to the region under the A ring due to the extended bulk of the biphenyl group. PMID- 11916147 TI - Evaluation of 7-hydroxy-flavones as inhibitors of oestrone and oestradiol biosynthesis. AB - A series of 4-aryl substituted 7-hydroxy-flavones were prepared using the three step Baker-Venkataraman synthesis in good overall yields. The flavones were all evaluated in vitro for inhibitory activity against aromatase (P450AROM, CYP19), using human placental microsomes, and for inhibitory activity against 17beta hydroxysteroid dehydrogenase type 1 (17beta-HSD-1) using human placental cytosol. The phenyl, 4-fluoro-phenyl and 4-bromo-phenyl derivatives displayed moderate inhibitory activity against P450AROM (IC50 17.2, 13.5 and 10.1 microM, respectively), none of the flavones, including the standard genistein, displayed any inhibitory activity against 17beta-HSD type 1 at 100 microM concentration. PMID- 11916148 TI - Antimycobacterial activity of 3,4-dichlorophenyl-ureas, N,N-diphenyl-ureas and related derivatives. AB - Substituted urea derivatives were prepared by reacting 3,4-dichlorophenyl isocyanate with amino acids, dipeptides, histamine or dicyandiamide among others, or from N,N-diphenyl-carbamoyl chloride and amino acids, dipeptides, or histamine. Other derivatives were obtained by reaction of PABA or PAS with arylsulfonyl halides. Some of the new compounds showed appreciable activity as antimycobacterial agents against Mycobacterium tuberculosis H37Rv, producing an inhibition of growth in the range of 80-89%, at a concentration of 6.25 microM. Some derivatives of this series might constitute interesting lead molecules for designing novel types of drugs effective against M. tuberculosis, a re-emerging pathogen both in the developed and under-developed countries. PMID- 11916149 TI - Phospholipase A2 from Trypanosoma brucei gambiense and Trypanosoma brucei brucei: inhibition by organotins. AB - Activity and kinetics of phospholipase A2 (PLA2) from Trypanosoma brucei gambiense (Wellcome strain) and Trypanosoma brucei brucei (GUTat 3.1) were examined using two different fluorescent substrates. The activity in the supernatants of sonicated parasites was Ca2+-independent, strongly stimulated by Triton X-100 with optimum activity at 37 degrees C and pH 6.5-8.5. To encourage a possible interaction between the parasite enzyme and organotin compounds, fatty acid derivatives of dibutyltin dichloride were synthesized and evaluated as potential inhibitors of PLA2. The enzyme from the two-trypanosome species differ with respect to kinetic parameters and are noncompetitively inhibited by the organotin compounds. The Michaelis constant (KM) for PLA2 from T. b. brucei is 63.87 and 30.90 microM while for T. b. gambiense it is 119.64 and 32.91 microM for the substrates 1,2-bis-(1-pyrenebutanoyl)-sn-glycero-3-phosphocholine (PBGPC) and 2-(12-(7-nitrobenz-2-oxa-1,3-diazol-4-yl)amino)dodecanoyl-1-hexadecanoyl-sn glycero-3-phosphocholine (NBDC12-HPC), respectively. PMID- 11916150 TI - Allosteric inhibition of rat liver and kidney arginase by copper and mercury ions. AB - Two isozyme forms of arginase are found in the rat. All arginases are metalloenzymes which require manganese for activity. Many arginases are activated by cobalt and nickel ions and inhibited by heavy metal ions. The purpose of this study was to compare the effect of other heavy metal ions on the rat liver isozyme (arginase I) and the rat kidney isozyme (arginase II). The activation and inhibition of arginase I and II by metal ions were different. However, both isozymes were strongly inhibited by cupric and mercuric ions. The inhibition of arginase I by cupric and mercuric ions was increased greatly by preincubation of the enzyme with the metal ions. However, preincubation of arginase II by cupric and mercuric ions had little effect on the inhibition of the enzyme. Under certain conditions the kinetics of the inhibition of both arginases I and II by cupric and mercuric ions was nonlinear allosteric. PMID- 11916151 TI - Protective effects of suprofen and its methyl ester against inactivation of rabbit kidney carbonyl reductase by phenylglyoxal. AB - Suprofen (SP) was little reduced by rabbit kidney carbonyl reductase, whereas its methyl ester (SPM) was an efficient substrate of the enzyme. To account for the differential catalytic activities for SP and SPM, the protective effects of these compounds against the inactivation of the enzyme by phenylglyoxal (PGO) were compared. Since the carboxyl group of SP is negatively charged and one essential arginine residue is known to be located in the NADPH-binding site of the enzyme, the protection of SP against the inactivation of the enzyme by PGO is expected to be more effective than that of SPM lacking a carboxyl group. However, the protective effects of SP and SPM were very similar. These results suggest that in spite of evidence for the binding of SP to the coenzyme-binding site, the carboxyl group of SP fails to interact with one essential arginine residue located in the site. PMID- 11916152 TI - Modification of human erythrocyte pyruvate kinase by an active site-directed reagent: bromopyruvate. AB - Human erythrocyte pyruvate kinase was modified with bromopyruvate and the kinetic behavior of the modified enzyme was investigated. When the enzyme was modified with bromopyruvate in the absence of adenosine-5'-diphosphate, phosphoenolpyruvate or fructose-1,6-diphosphate the inactivation followed a pseudo first-order kinetics. The inactivation rate constant, ks, was 1.84 +/- 0.15 min(-1). Kd of the bromopyruvate-enzyme complex was 0.14 +/- 0.03 mM. The presence of adenosine-5'-diphosphate, phosphoenolpyruvate or fructose-1,6 diphosphate in the modification medium or the presence of fructose-1,6 diphosphate in the assay medium resulted in deviation of the inactivation kinetics from pseudo first-order. Phosphoenolpyruvate was better than adenosine 5'-diphosphate for protection against bromopyruvate modification whereas fructose 1,6-diphosphate was ineffective. The modified enzyme showed negative cooperativity in the presence of fructose-1,6-diphosphate whereas in the absence of it no activity was detected. PMID- 11916153 TI - The advanced adenoma as the primary target of screening. AB - The advanced adenoma bridges benign and malignant states and may be the most valid neoplastic surrogate marker for present and future colorectal cancer risk. We define the advanced adenoma as an adenoma with significant villous features (>25%), size of 1.0 cm or more, high-grade dysplasia, or early invasive cancer. Prevention studies should demonstrate a high efficacy in reducing the number of advanced adenomas. We should use the advanced adenoma in the evaluation of new screening technology, nutritional interventions, and chemoprevention agents because the advanced adenoma is a more desirable target for screening efficacy than is the more uncommon but life-threatening cancer stage or the more common but early, less significant small adenoma stage. PMID- 11916154 TI - Fecal occult blood test screening for colorectal cancer. AB - In summary, high-quality scientific studies indicate that the use of the FOBT for colorectal cancer screening has a number of important advantages. The test is capable of detecting most early colorectal cancers and many advanced adenomas. It has been shown in randomized, controlled trials to reduce substantially colorectal cancer mortality and incidence. The FOBT is feasible, widely available, and acceptable to most individuals. It has a low up-front cost and is highly cost-effective. Combining annual FOBT with periodic flexible sigmoidoscopy seems to be an especially effective screening option. Limitations of FOBT screening include its low sensitivity for polyps, especially smaller ones. Some of the trials report a relatively low sensitivity for detecting cancers located in the distal colon. The test has a relatively low specificity, so there are many false-positive screens; and for it to be most effective, repetitive screening is necessary. Balancing these advantages and disadvantages, the evidence-based screening guidelines have concluded that FOBT screening has a major role to play in colorectal cancer control and a program of annual FOBT plus flexible sigmoidoscopy every 5 years is a preferred option for screening the asymptomatic, average-risk population for colorectal cancer. Short of doing direct colonoscopy screening for the entire at-risk population, the FOBT currently is the best available method of identifying asymptomatic, average-risk people most likely to benefit from colonoscopy. PMID- 11916155 TI - Familial and inherited colorectal cancer: endoscopic screening and surveillance. AB - Familial risk of colorectal cancer is very common. The high-risk inherited syndromes are well described and much is known about the genetics and the effectiveness of registration, endoscopic surveillance, and appropriate intervention in these patients. The inherited syndromes, however, are extremely rare. There is a large group of patients in our population who can benefit from risk stratification based on the number of their relatives with colon cancer or adenomas and the age at which those relatives developed neoplasm. The GI endoscopist has a vital role in recommending and providing colonoscopic screening for this large group of patients. PMID- 11916156 TI - Colorectal cancer surveillance for patients with inflammatory bowel disease. AB - Although the cumulative prevalence of colorectal cancer among patients with ulcerative colitis is similar to that among patients with sporadic colorectal cancer, the younger age of ulcerative colitis patients with cancer accounts for the age-specific relative risk. Approximately half of ulcerative colitis patients with colorectal cancer die from metastatic disease. Pancolitis of long duration and coexistent primary sclerosing cholangitis are strong risk factors for cancer that should prompt entry into cancer surveillance programs. When done appropriately, cancer surveillance in patients with inflammatory bowel disease can be very effective, and at a reasonable cost. PMID- 11916157 TI - Barriers to screening for colorectal cancer. AB - Rapidly growing interest in colon cancer screening is a crucial first step to identifying and reducing many of the barriers that impede population screening for this common disease. Promoting screening demands health care policy change to increase the percentage of Americans with insurance coverage that includes a colon cancer screening benefit. A systematic approach to screening with invitations that come from a clinician are likely to be the most effective way to prompt more individuals to be screened. Awareness campaigns and patient educational aids, including decision tools, implemented in multiple sites, such as worksites, community centers, health care systems, and physician offices, increase the percent of eligible Americans who understand their personal risk, the need for screening, and the options available to them. PMID- 11916158 TI - Medicolegal implications of colon cancer screening. AB - The treatment of colon cancer is a significant source of malpractice liability. Physicians are justifiably concerned about malpractice exposure as personal injury attorneys investigate standard and novel malpractice theories and claims. With the general acceptance of the importance of screening for colon cancer, screening for colon cancer is now defined as the standard of care. This has opened up a previously neglected area of malpractice liability. Physicians need to understand the sources of malpractice risk and risk-management strategies related to these sources to reduce their exposure to liability suits in this area. This article outlines these sources evolving from the tort of negligence, including the duty to provide care, practicing below the standards of care, the cause of the harm, and the actual establishment of harm. The concept of vicarious liability and its relationship to the tort of negligence also are discussed. This presentation is developed within the context of a risk-management approach to assist physicians in developing a preventive approach to malpractice liability. PMID- 11916159 TI - Flexible sigmoidoscopy: an important screening option for average-risk individuals. AB - Colorectal cancer screening techniques should be effective, acceptable to patients, affordable, widely available, and safe. For average-risk adults aged more than 50 years who do not have significant colorectal symptoms, significant family history, or significant predisposing conditions, flexible sigmoidoscopy is an important option for reducing the risk for colorectal cancer, meeting all criteria for an effective and feasible screening modality. This article discusses evidence supporting flexible sigmoidoscopy, practical issues in implementation, and current controversies. PMID- 11916160 TI - Clinical significance of small polyps found during screening with flexible sigmoidoscopy. AB - In deciding how to interpret the significance and management of small distal adenomatous polyps found on FS, one must first decide on the goal of a screening program. If the goal is maximal reduction of CRC risk, regardless of cost, there is little argument that screening colonoscopy is the most effective approach. Unfortunately cost and cost-effectiveness are important considerations when administering a screening program with a fixed budget. Although comparing the cost-effectiveness of different strategies is beyond the scope of this article, rigorous comparisons by other authors have suggested that a sigmoidoscopy-based approach is more cost-effective than a colonoscopy-based approach. The most cost effective approach may change, however, if the frequency of screening and surveillance can be reduced without significantly impacting effectiveness. Other authors using assumptions including low compliance rates for regular FOBT or FS have determined that colonoscopy every 10 years is the most cost-effective approach. Multiple studies support the recommendation that villous polyps regardless of size and adenomatous polyps greater than 1 cm found on FS are important markers for the presence of advanced polyps and cancer in the proximal colon. These patients should undergo colonoscopy. If one assumes that a sigmoidoscopy-based approach is reasonable, and accepts that such an approach always misses a small number of proximal lesions, how should one manage patients with a small adenomatous polyp on FS? In aggregate, the studies discussed previously suggest that patients with no distal polyps, distal hyperplastic polyps, or a single small distal tubular adenoma have a similar and low risk of advanced proximal adenomas of the colon. There are some studies, however, that do not support this. With the exception of the study by Read et al, these studies included patients at elevated risk of CRC because of a family history, or inclusion of patients with positive FOBT (or not tested). The study by Read et al also included patients with distal villous adenomas in their low-risk group. Because a sigmoidoscopy-based strategy typically excludes patients at elevated risk, these results may not be applicable to low-risk patients undergoing sigmoidoscopy. Given these caveats, what can one conclude about the predictive value of a small tubular adenoma found on FS? These studies suggest that the risk of proximal advanced polyps is similar or slightly increased in patients with a distal adenoma than those with a negative FS. The risk of finding an advanced adenoma seems to be 0% to 4% regardless of the findings of no polyps, hyperplastic polyps, or small tubular adenomatous polyps on FS in low-risk patients. A small portion of patients with hyperplastic polyps found on FS have advanced proximal adenomas. If a hyperplastic polyp on FS is not an indication for colonoscopy and the risk of proximal advanced adenomas is similar in patients with only a small distal adenoma, it is inconsistent to recommend colonoscopy for a small distal tubular adenoma and not a hyperplastic polyp. Based on the studies of asymptomatic patients with no family history and negative FOBT, the authors believe it is reasonable to defer colonoscopy if no polyp, a hyperplastic, or a small tubular adenoma is found at sigmoidoscopy in low-risk patients. If the patient or physician is unwilling to accept a small (0% to 4%) chance of missing an advanced proximal lesion, then a sigmoidoscopy-based approach (regardless of the threshold to go on to colonoscopy) is not appropriate. Screening FS remains an effective examination to screen for CRC in asymptomatic patients. There is no question that colonoscopy clearly detects more lesions than FS. It remains to be seen if the increase in costs and risks justifies the improved detection rate of colonic polyps. Given manpower issues that face us today, and examining the question from a population perspective, reserving colonoscopy for only those patients with an advanced distal polyp on FS gives the biggest yield. PMID- 11916161 TI - Rationale for combining different screening strategies. AB - It is generally accepted that screening programs should be quite safe, and that the benefits should substantially outweigh the harms. As Cochrane and Holland stated: We believe that there is an ethical difference between everyday medical practice and screening. If a patient asks a medical practitioner for help, the doctor does the best he can. He is not responsible for defects in medical knowledge. If, however, the practitioner initiates screening procedures he is in a very different situation. He should, in our view, have conclusive evidence that screening alters the natural history of disease in a significant proportion of those screened. If this is so, one should recommend the combination over either test alone only if there is sufficient evidence that the combination is more effective and no more dangerous. There is a difference of opinion over whether the evidence, which is certainly not strong, is nevertheless sufficient. This poses a dilemma. Many expert groups prefer that screening for colorectal cancer be done with both FOBT and sigmoidoscopy rather than either alone. Yet, the strength of the evidence for additional effectiveness, and information on the magnitude of that effect if it is present, is substantially less than for the individual tests. This being the case, the author believes that it is premature to advocate the combination over either test alone, especially when the most pressing national priority in colorectal cancer screening is to get a large proportion of the adult population to be screened at all. PMID- 11916162 TI - Rationale for colonoscopy screening and estimated effectiveness in clinical practice. AB - Colonoscopy screening has the highest anticipated level of effectiveness of the available colorectal cancer screening techniques. Its long-term cost effectiveness is also comparable with or superior to other modalities. Evidence for the expected reduction in colorectal cancer incidence and mortality varies with colonoscopy screening from 50% to 90%, for reasons that are not fully understood. Maintaining a high standard of performance is critical with regard to achieving the highest level of effectiveness possible. PMID- 11916163 TI - Technical assessment of direct colonoscopy screening: procedural success, safety, and feasibility. AB - Colonoscopy, when performed by adequately trained physicians, is a safe and effective procedure for colorectal cancer screening. To realize the benefits of colonoscopic screening of the general population for colorectal cancer, it is imperative that physicians performing this procedure receive appropriate training to maintain the highest standards of patient care. PMID- 11916164 TI - Does virtual colonoscopy have a major role in population-based screening? AB - In summary, technical advances in the performance of VC are occurring at a very rapid pace. These technical improvements will undoubtedly improve the polyp detection rate and reduce false-positive and false-negative examinations. The concept of VC is clearly attractive and the general public is enamored with everything that has an association with virtual reality. As other articles in this issue have revealed, there are many techniques in development to help stratify patients at risk for colon cancer. As we begin to focus our health care resources on those at highest risk, the less need there is for inexpensive, broadly based screening techniques. Clearly, those patients at high risk for having polyps are better served by colonoscopy because of its therapeutic potential. That being said, in the view of this author, if a virtual preparation can be achieved and the cost of VC can be kept relatively low, then this technique will become part of our mainstream clinical practice. If an immaculate colon preparation must be performed and if the costs reflect standard abdominal and pelvic CT rather than a special reduced cost for VC, then it is doubtful that there will be any significant impact from this technology PMID- 11916165 TI - Cost-effectiveness of screening the average-risk population for colorectal cancer. AB - This article reviews several of the recent models addressing the cost effectiveness of colorectal cancer screening in the average-risk individual (Table 1). How can clinicians and policy makers use this information for decision making regarding colorectal cancer screening? The cost-effectiveness ratios reported by themselves do not identify cost-effective practices. They must be placed in a decision context that is expressed in one of two forms. In the first form, an explicit threshold or maximum amount that a policy maker is willing to spend is stated (e.g., $40,000 per LY gained, as has been quoted as an acceptable amount for a prevention program). In the second form of decision context, a list of medical practices and their associated cost-effectiveness ratios, also known as a league table (Table 2) is used as a basis for comparison with the practice under evaluation (e.g., colorectal cancer screening). The practice with the lowest cost-effectiveness ratio is the most cost-effective practice on the list. Practices with lower cost-effectiveness ratios are considered cost-effective compared with those with higher ratios. Table 2 lists incremental cost effectiveness ratios for common medical practices. The models discussed in this article suggested that colorectal cancer screening using annual FOBT, flexible sigmoidoscopy at 3 or 5 years, the combination of FOBT and flexible sigmoidoscopy, barium enema, colonoscopy, and even virtual colonoscopy had incremental cost-effectiveness ratios ranging from $6300 to $92,900 per LY saved with most of the cost-effectiveness ratio ranging from $10,000 to $40,000 per LY saved. These ratios are similar to the cost of another widely accepted practice, breast cancer screening with annual mammography in women age 50 and older ($22,000 per LY gained). Colorectal cancer screening with any of the modalities discussed is considered less cost-effective than screening for hemochromatosis, which has an incremental cost-effectiveness ratio of $3665 per LY saved. Based on these ratios, however, screening for colorectal cancer is considered cost effective compared with cervical cancer screening in women age 20 and older with pap smear every 3 years, which has an incremental cost-effectiveness ratio of $250,000 per LY gained. The clinician can use these incremental cost effectiveness ratios to evaluate the risks and benefits of alternative practices for the individual, and the policy maker with a limited health care budget can use these ratios to set priorities for funding based on the costs and the expected gains in life expectancy for colorectal cancer screening and for alternative health care programs. PMID- 11916166 TI - A novel CD28 mRNA variant and simultaneous presence of various CD28 mRNA isoforms in human T lymphocytes. AB - The primary transcript of the human CD28 gene in T lymphocytes, encoding for a costimulatory molecule, is known to undergo alternative splicing, and different small sets of variant isoforms have been reported. This report presents the novel simultaneous presence of eight different mRNA isoforms, all observed together in normal human T cells; this is an interesting finding in the study of CD28 mRNA structural variants. A similar pattern was found in a total of four individuals. In addition, we also report the occurence and sequence of a new CD28 mRNA isoform, one of the above eight, which is a novel variant generated by the use of a new combination of splice donor and acceptor sites. PMID- 11916167 TI - Heterogeneity of HLA-DRB1*04 and its associated haplotypes in the North Indian population. AB - HLA-DR4 has been implicated in several diseases including rheumatoid arthritis (RA) and type I diabetes, the strength of associations being ethnically variable. Unusually high level of heterogeneity in DR4-DQB1 haplotypes has been reported in the Indian population. The present study is an attempt to determine the genetic diversity of the HLA-DR4 allelic family and its associated DQA1-DQBI haplotypic combinations in the healthy North Indian population. Using PCR-SSP and PCR-SSOP techniques, nine subtypes of DR4 were encountered of which DRB1*0403 was the most predominant allele (34.8%) followed by *0404 (27%), *0401 (14.6%), and *0405 (11%). No examples of *0402, *0409, *0411, *0413-*0417, and *0419-23 were encountered, although a few other subtypes, *0410 (three examples), *0406 and *0418 (two examples each), and *0407, *0408, and *0412 (single example each) occurred infrequently in a cohort of 85 HLA-DR4 positive samples studied. Most of these subtypes occurred in combination with DQA1*03-DQB1*0302 (69.5%). DRB1*0403 and *0404 exhibited maximum heterogeneity of DQB1 combinations. Haplotype data revealed the presence of 15 different DR4-DQ haplotypes, four of which were found to be "unique" to Asian Indians, not reported in any other population. These results help to explain the observed variability in DR4 associations in autoimmune diseases in Asian Indians and provide support for scientific and historical documentation of extensive admixture in the Indian subcontinent. PMID- 11916168 TI - MICA is a target for complement-dependent cytotoxicity with mouse monoclonal antibodies and human alloantibodies. AB - The highly polymorphic major histocompatibility class I related chain A (MICA) gene encodes glycoproteins that have been shown to be expressed in epithelial cells, endothelial cells, keratinocytes, monocytes, and tumor cells. In previous experiments, we have studied MICA antigens using rabbit sera obtained by immunization with MICA peptides. We also found that several transplant recipients had specific antibodies against MICA in an ELISA assay with recombinant of MICA (r-MICA). In the present work we produced monoclonal antibodies by immunization of mice with recombinant MICA*008. Based on the different patterns of reactivity observed in ELISA, Western blot, and flow cytometry, mAbs 1.9C2, 2.4F5, 1.7AD, and 2.3D4 only reacted with denatured MICA and mAb 1.7A8 and 3.2H3 reacted also with native MICA as illustrated by flow cytometry with live cells. These monoclonal antibodies were postulated to bind to different sites of the MICA molecule. In order to investigate whether MICA expressed on the cell surface is able to mediate cell killing, antibody absorption, flow cytometry and complement dependent cytotoxicity (CDC) were performed. We found that mouse monoclonal antibody 3.2H3 was able to kill 70% of HeLa cells. Absorption of a patient serum with pooled human platelets to remove antibodies against class I HLA resulted in a small shift of fluorescence and reduced killing from 100% to 70-75%. Absorption with the platelets and r-MICA produced a remarkable reduction in fluorescence staining and virtually reduced complement-dependent killing to the level of the negative controls. The results suggested that MICA alloantigens may be more immunogenic than could have been previously suspected. PMID- 11916169 TI - Relative and absolute HLA-DQA1-DQB1 linked risk for developing type I diabetes before 40 years of age in the Belgian population: implications for future prevention studies. AB - HLA-DQ genotyping remains the cornerstone of genetic risk stratification in type I diabetes prediction and prevention studies. We developed a genetic screening strategy for predisposition to type I diabetes in the Belgian population based upon HLA-DQA1-DQB1 typing and taking into account the age at clinical onset. A group of 1866 autoantibody-positive type I patients below age 40 years recruited by the Belgian Diabetes Registry and a group of 750 control subjects were DQA1 DQB1 genotyped. In the total study population 16 different DQA1-DQB1 haplotypes were revealed, allowing the stratification of 81 genotypes in ten different genotype groups. Apart from the highest risk DQA1*-DQB1* genotype 0301-0302/0501 0201 (odds ratio 21; absolute risk 6%), three other genotype groups conferred a highly significant disease risk (p < 10(-6)). Altogether, these susceptibility genotypes were carried by 9% of the control subjects versus 60% of the patients diagnosed before age 40 years and up to 70% of those under age 5 years. All other genotypes were protective, neutral, infrequent or associated with a moderate protection or susceptibility. A strong, although not absolute protection was conferred by DQB1*0602-positive haplotypes (odds ratio = 0.03). This study in a large cohort of autoantibody-positive patients shows that a DQA1-DQB1-based genotyping strategy allows the identification of a subgroup representing less than 10% of the Belgian population but harbouring the majority of future type I patients arising in childhood or early adulthood. Future prediction and prevention studies should take into account the age dependency of this HLA-DQ associated risk. PMID- 11916170 TI - TCRBV CDR3 diversity of CD4+ and CD8+ T-lymphocytes in HIV-infected individuals. AB - TCRBV CDR3 repertoire diversity was analyzed in a cross-sectional study of HIV infected individuals by CDR3 fingerprinting/spectratyping and single strand conformation polymorphism (SSCP). Most TCRBV families were detected in CD4+ cells of HIV-infected patients with CD4 counts ranging from 35 to 1103. In patients with CD4 counts >500, CD4+ TCRBV CDR3 fingerprinting profiles contained subtle variations with generally gaussian-distributed sizes. Lower CD4 counts coincided with more fragmented TCRBV CDR3 repertoires, containing dominant bands and bands missing from the CDR3 profiles. The CD8+ population of the same patients exhibited skewed CDR3 profiles of the majority of TCR BV families at CD4 counts >500. Irregularity of CD8+ CDR3 size distribution was most profound at low CD4 counts and suggested domination of the CD8+ TCRBV repertoire by a limited number of clones. Skewed patterns of CDR3 diversity probably reflect (oligo)clonal expansion of particular CD4+ and CD8+ cell populations during chronic infection with HIV. In addition, irregular CDR3 profiles of CD4+ and CD8+ at low CD4 counts suggest diminished TCR repertoire diversity, which may contribute to immunodeficiency. PMID- 11916171 TI - Analysis of TAP2 polymorphisms in Finnish individuals with type I diabetes. AB - Type I diabetes mellitus is an immune-mediated disease that is known to be associated and linked with genes in the human leukocyte antigen (HLA) region on chromome 6. Functionally, HLA class I antigen presentation may be deranged in type I diabetes. The TAP1 and TAP2 transporters, which mediate the translocation of antigenic peptides into the endoplasmic reticulum and whose genes are located in the HLA class II region, are potential candidates for conferrring predisposition to type I diabetes. Five known coding region variants (codons 379, 565, 651, 665, and 687) as well as three new polymorphisms of TAP2, one silent (codon 604) and two intronic (nucleotide positions 49,270 and 49,471), were typed in a cohort of 146 well-characterized Finnish individuals with type I diabetes and 90 control subjects. Absolute linkage disequilibrium was apparent for the polymorphisms at codons 604, 665, and 687 as well as the two downstream intronic polymorphisms in a 613-bp region of the 3' portion of TAP2; the polymorphism at codon 651, which is also present within this region, was excluded from this linkage. The codon 651 polymorphism defines the allele TAP2F, the frequency of which in HLA-DR4+ diabetic subjects was 5.4 times that in DR4+ controls (27 vs. 5%, p = 0.002, p(c) = 0.01). These data are consistent with the existence of susceptibility haplotypes for type I diabetes in the Finnish population consisting of DRB1*04 (*0401 and *0404), DQ8, and TAP2F. PMID- 11916172 TI - Association of the CTLA4 3' untranslated region polymorphism with the susceptibility to rheumatoid arthritis. AB - Cytotoxic T-lymphocyte antigen 4 (CTLA4) gene polymorphism located in the 3' untranslated region (UTR) was investigated in 141 Spanish patients (38 men and 103 women) with rheumatoid arthritis (RA) and in 194 ethnically-matched healthy controls. Twenty alleles having different numbers of (AT) repeats (from 7 to 32) were found in this population. (AT)7 and (AT)16 were the most frequent alleles, and accounted for almost two-thirds of the allelic frequency in the control population. Consequently, alleles were assigned as L (large: 16 or more AT repeats) or S (short: less than 16 AT repeats). When the L/S distribution in patients and controls were compared, an increase of L alleles was observed among patients (49.9% vs. 39.7%; p = 0.02; p(c) = 0.04, odds ratio [OR] = 1.46; 95% confidence interval [CI], 1.06-2.01). Hence, the frequency of S alleles was decreased among patients (51.1% vs. 60.3%; p = 0.02; p(c) = 0.04; OR = 0.69; 95%CI, 0.50-0.95). Moreover, a statistically significant decrease in the frequency of S/S individuals was observed among RA patients (27.7% versus 40.7%; p = 0.01; p(c) = 0.03; OR = 0.56; 95%CI, 0.34-0.91). These differences were irrespective of the HLA "shared epitope" (SE) status, and were observed similarly among SE+ as well as among SE- patients. After combining these data with other reported previously by us, from studies of CTLA4 49 (A/G) and -318 (C/T) polymorphisms, we conclude that the strongest association between CTLA4 gene polymorphisms and RA susceptibility occurs with the 3' UTR polymorphism. PMID- 11916173 TI - Allogeneic MHC class I ligands and their role in positive and negative regulation of human cytotoxic effector cells. AB - The allogeneic mixed lymphocyte culture (MLC) has served as an important experimental system for elucidating the cellular and molecular basis of human lymphocyte responses. Complex mixtures of lymphocytes are stimulated by disparate alloantigens, inducing cellular activation and generating a cytokine milieu that is an excellent breeding ground for the proliferation and differentiation of many distinct lymphocyte subsets. Cloning of individual lymphocytes following alloactivation has allowed various cytotoxic lymphocytes to be isolated and characterized with respect to phenotype and specificity. These analyses have revealed that all types of cytotoxic effector cells are regulated by interactions with MHC-peptide ligands, however, the consequences of these interactions can result in opposite functional outcomes. In this review we summarize how allogeneic MHC class I-peptide ligands positively or negatively regulate the activities of four distinct groups of cytotoxic lymphocytes and how this information might be transferred into clinical use. PMID- 11916174 TI - Influenza and current guidelines for its control. PMID- 11916176 TI - Virulence factors of Escherichia coli isolated from female reproductive tract infections and neonatal sepsis. AB - OBJECTIVE: The presence of enterobacteria such as Escherichia coli in the vagina of normal women is not synonymous with infection. However, vaginal E. coli may also cause symptomatic infections. We examined bacterial virulence properties that may promote symptomatic female reproductive tract infections (RTI) and neonatal sepsis. METHODS: E. coli isolated as the causative agent from cases of vaginitis (n = 50), tubo-ovarian abscess (n = 45) and neonatal sepsis (n = 45) was examined for selected phenotypic and genetic virulence properties. Results were compared with the frequency of the same properties among fecal E. coli not associated with disease. RESULTS: A significantly greater proportion of infection E. coli exhibited D-mannose resistant hemagglutination compared with fecal E. coli (p < 0.01). This adherence phenotype was associated with the presence of P fimbriae (pap) genes which were also significantly more prevalent among isolates from all three infection sites (p < 0.01). The majority of pap+ isolates contained the papG3 allele (Class II) regardless of infection type. Increased frequency of Type IC genes among vaginitis and abscess isolates was also noted. No significant differences in frequency of other bacterial adherence genes, fim, sfa, uca (gaf or dra were observed. E. coli associated with vaginitis was significantly more likely to be hemolytic (Hly+) than were fecal isolates (p < 0.05). The Hly+ phenotype was also more prevalent among tubo-ovarian abscess and neonatal sepsis isolates (p < 0.08). CONCLUSIONS: E. coli isolated from female RTI and neonatal sepses possess unique properties that may enhance their virulence. These properties are similar to those associated with other E. coli extra-intestinal infections, indicating that strategies such as vaccination or bacterial interference that may be developed against urinary tract infections (UTI) and other E. coli extra-intestinal infections may also prevent selected female RTI. PMID- 11916175 TI - A randomized trial of azithromycin versus amoxicillin for the treatment of Chlamydia trachomatis in pregnancy. AB - OBJECTIVE: To compare the compliance, side effects and efficacy of amoxicillin and azithromycin for the treatment of Chlamydia trachomatis infection in pregnancy. METHODS: This is a randomized single-blind trial of women diagnosed with C. trachomatis before 33 weeks gestation. Women were randomlyassigned either 500 mg amoxicillin orally three times per dayfor 7 days or a single dose of 1 g azithromycin orally. Patients were interviewed by telephone approximately 3-7 days following therapy to assess compliance and side effects. Test of cure was performed at a follow-up visit 4-6 weeks following completion of therapy. RESULTS: Thirty-nine patients were randomized with 19 receiving amoxicillin and 20 receiving azithromycin. There were no differences in baseline data between the two groups, and there were no statistically significant differences in side effects, compliance or efficacy. In the amoxicillin group 84% of women took all pills, while 100% completed the single 1 g dose of azithromycin. Side effects were common in both groups (38% overall), with 40% of the azithromycin group reporting moderate to severe gastrointestinal side effects compared to 17% in the amoxicillin group (p = 0.11). Of patients who returned for follow-up test of cure, 3 of 15 (20%) in the amoxicillin group were positive compared with 1 of 19 (5%) in the azithromycin group (p = 0.3). CONCLUSIONS: Side effects of therapy for C. trachomatis in pregnancy are common. Amoxicillin was slightly better tolerated than azithromycin. Compliance and cure rates with both regimens was high. PMID- 11916177 TI - Performance characteristics of putative tests for subclinical chorioamnionitis. AB - OBJECTIVE: To evaluate amniotic fluid glucose, matrix metalloproteinase (MMP)-9, interleukin (IL)-6, and IL-12 for diagnosing subclinical chorioamnionitis in women with preterm labor. METHODS: Forty-four women in preterm labor at 22-35 weeks gestation with suspected subclinical chorioamnionitis underwent amniocentesis. Amniotic fluid analysis included Gram stain, culture, and determination of glucose, MMP-9, IL-6, and IL-12 concentrations. Median values of these analytes were compared using the Mann-Whitney U test. Sensitivity, specificity, and positive and negative predictive values were calculated for tests using a positive amniotic fluid culture or delivery within 24 hours as the key outcome variables. RESULTS: Amniotic fluid concentrations of glucose, MMP-9, and IL-6 correlated closely with positive culture or delivery within 24 hours. IL 12 concentrations did not correlate with either a positive culture or delivery within 24 hours. CONCLUSIONS: Amniotic fluid glucose, MMP-9, and IL-6 reliably predict microbial invasion of the amniotic cavity or imminent delivery. IL- 12 values did not correlate with amniotic fluid culture results or imminent delivery. PMID- 11916179 TI - Differentiation between women with vulvovaginal symptoms who are positive or negative for Candida species by culture. AB - OBJECTIVE: To investigate whether clinical criteria could differentiate between women with vulvovaginitis who were culture positive or negative for vaginal Candida species. METHODS: Vulvovaginal specimens were obtained from 501 women with a vaginal discharge and/or pruritus. Clinical information and wet mount microscopy findings were obtained. All specimens were sent to a central laboratory for species identification. RESULTS: A positive culture for Candida species was obtained from 364 (72.7%) of the specimens. C. albicans was identified in 86.4% of the positive cultures, followed by C. glabrata in 4.5%, C parapsilosis in 3.9%, C. tropicalis in 2.7% and other Candida species in 1.4%. Women with a positive Candida culture had an increased utilization of oral contraceptives (26.1% vs. 16.8%, p = 0.02) and antibiotics (8.2% vs. 0.7%, p = 0.001), and were more likely to be pregnant (9.1% vs. 3.6%, p = 0.04) than the culture-negative women. Dyspareunia was more frequent in women without Candida (38.0% vs. 28.3%, p = 0.03) while vaginal erythema (p = 0.01) was more common in women with a positive Candida culture. CONCLUSIONS: Although quantitative differences were observed, the presence of vaginal Candida vulvovaginitis cannot be definitively identified by clinical criteria. PMID- 11916178 TI - Prevalence of human immunodeficiency virus, hepatitis B and hepatitis C virus antibodies and hepatitis B antigen among commercial sex workers in Japan. AB - OBJECTIVE: To investigate the prevalence of antibodies to human immunodeficiency virus (HIV), hepatitis B virus (HBV) and hepatitis C virus (HCV), and of hepatitis B surface (HBs) antigen in commercial sex workers (CSW) who attended a sexually transmitted disease (STD) clinic in Tokyo. METHODS: Surveys were conducted on 308 CSW and 384 control subjects for HIV antibody or 241 control subjects for HBs antibody and antigen and HCV antibody. RESULTS: HIV antibody was not detected in either CSW or control subjects. The positive rates for HBs antigen and antibody were 0.6 and 23.4%, respectively, in the CSW group, and 0.4 and 71.8% in the control group. The HCV antibody positive rate was 3.2% in the CSW group and 0.4% in the control group. CONCLUSION: A statistically significant difference between the two groups was observed only in HCV antibody positive rate. STD checkup for CSW alone is inadequate--STD health education and screening for the general public are also required. PMID- 11916180 TI - Impaired whole-blood polymorphonuclear leukocyte migration as a possible predictive marker for infections in preterm premature rupture of membranes. AB - OBJECTIVES: Steroids, used in preterm premature rupture of membranes (pPROM), to reduce the risk of morbidity and mortality of the preterm neonate, impair the maternal polymorphonuclear leukocyte (PMN)-based immune system. In spite of combination with antibiotics, prenatal and postnatal bacterial infections of mother and child are frequent. This pilot study focuses on the influence of steroids in pPROM on maternal PMN functional capacity and subsequent infections. METHODS: After opting for expectant management, eight women with pPROM and no signs of infection were treated by steroids (betamethasone 5.7 mg, i.m. every 24 hours, for three days) and antibiotic therapy with either amoxicillin and clavulanic acid, piperacillin or ampicillin i.v. up to delivery. The conventional inflammation parameters of PMN blood count and C-reactive protein (CRP) were measured daily in parallel with PMN migratory capacity towards N-formyl-methionyl leucyl-phenylalanine stimulation and under blank conditions, estimated by a whole blood membrane filter assay. RESULTS: In all patients PMN migration decreased during the application of steroids. Three patients showed a decrease in PMN migration below critical values and in spite of antibiotic prophylaxis acute pyelonephritis developed 2-6 days later. PMN count and CRP were not predictive of maternal infection. CONCLUSION: Reduced PMN function, caused by steroid treatment in pPROM, is suggested to be a reason for serious bacterial infections in spite of antibiotic prophylaxis. PMN migration reflects individual PMN defensive capacity. PMID- 11916181 TI - A randomized controlled trial of interleukin-1 receptor antagonist in a rabbit model of ascending infection in pregnancy. AB - OBJECTIVE: To determine whether treatment with interleukin-1 receptor antagonist (IL1-ra) would affect amniotic fluid concentrations of tumor necrosis factor alpha (TNF-alpha) and prostaglandins or clinical or microbiological outcomes in a model of ascending bacterial infection in pregnancy. METHODS: Timed pregnant New Zealand white rabbits at 70% of gestation underwent endoscopic inoculation of the cervices with 10(6) - 10(7) cfu Escherichia coli. Animals were randomly assigned in a blinded manner to a 5-h intravenous infusion of human IL1-ra (10 mg/kg) or placebo beginning 1-2 h after inoculation. Blood was drawn from the does for assay of serum IL1-ra concentration before inoculation, at mid-infusion, after the infusion ended and at necropsy. At necropsy, temperature and cultures were taken, and aspirated amniotic fluid was pooled for assays of TNF-aalpha, prostaglandin E2 (PGE2) and ILI-ra. RESULTS: Serum IL1-ra concentrations rose to a mean of 2 microg/ml at mid-infusion and fell markedly after the infusion to concentrations barely detectable at necropsy. Between the two groups, there were no significant differences in the rates of fever or positive cultures or in amniotic fluid concentrations of PGE2 or TNF-alpha. One unique finding was the demonstration that administration of human IL1-ra to the does resulted in measurable concentrations of human IL1-ra in the amniotic fluid. CONCLUSIONS: Treatment with an intravenous infusion of human IL1-ra after cervical inoculation with E. coli did not affect clinical or microbiological outcomes or amniotic fluid concentrations of TNF-alpha or PGE2. This experiment providesthefirstdemonstration of passage of human IL1-ra from the maternal bloodstream to the amniotic fluid. PMID- 11916182 TI - The inhibitory effect of clindamycin on Lactobacillus in vitro. AB - OBJECTIVE: To evaluate the in vitro effect of varying concentrations of clindamycin on Lactobocillus spp. METHODS: Concentrations of clindamycin ranging from 1.95-20,000 microg/ml were studied for their effect on the growth of six strains of Lactobacillus. RESULTS: Clindamycin concentrations between 1.95-31.25 microg/ml had no statistically significant effect on growth of lactobacilli (p > 0.05). Concentrations 125 and 250 microg/ml had a bacteriostatic effect. The mean minimum inhibitory concentration (MIC) for studied Lactobacillus strains was determined as 1,000 microg/ml. CONCLUSION: High concentrations of clindamycin achieved in the vagina by intravaginal application might be inhibitory for Lactobacillus. PMID- 11916183 TI - Candida lusitaniae as an unusual cause of recurrent vaginitis and its successful treatment with intravaginal boric acid. AB - Increasing use of short-course antifungal therapies in patients with recurrent vulvovaginitis may enable the emergence of less-common, more resistant yeast strains as vaginal pathogens. We report the case of a patient with chronically symptomatic and repeatedly treated vaginal candidiasis whose infection was attributable to Candida lusitaniae, a previously unreported cause of candidal vaginitis. PMID- 11916184 TI - Urinalysis and urinary tract infection: update for clinicians. AB - Dysuria is a common presenting complaint of women and urinalysis is a valuable tool in the initial evaluation of this presentation. Clinicians need to be aware that pyuria is the best determinate of bacteriuria requiring therapy and that values significant for infection differ depending on the method of analysis. A hemocytometer yields a value of > or = 10 WBC/mm3 significant for bacteriuria, while manual microscopy studies show > or = 8 WBC/high-power field reliably predicts a positive urine culture. In cases of uncomplicated symptomatic urinary tract infection, a positive value for nitrites and leukocyte esterase by urine dipstick can be treated without the need for a urine culture. Automated urinalysis used widely in large volume laboratories provides more sensitive detection of leukocytes and bacteria in the urine. With automated microscopy, a value of > 2 WBC/hpf is significant pyuria indicative of inflammation of the urinary tract. In complicated cases such as pregnancy, recurrent infection or renal involvement, further evaluation is necessary including manual microscopy and urine culture with sensitivities. PMID- 11916185 TI - Transjugular intrahepatic portosystemic shunt versus endoscopic therapy: randomized trials for secondary prophylaxis of variceal bleeding: an updated meta analysis. PMID- 11916186 TI - In vitro studies indicate that acid catalysed generation of N-nitrosocompounds from dietary nitrate will be maximal at the gastro-oesophageal junction and cardia. AB - BACKGROUND: Dietary nitrate increases saliva nitrite levels and swallowed saliva is the main source of nitrite entering the acidic stomach. In acidic gastric juice, this nitrite can generate potentially carcinogenic N-nitrosocompounds. However, ascorbic acid secreted by the gastric mucosa can prevent nitrosation by converting the nitrite to nitric oxide. METHODS: To study the potential for N nitrosocompound formation in a model simulating salivary nitrite entering the acidic stomach and the ability of ascorbic acid to inhibit the process. Concentrations of ascorbic acid, total vitamin C, nitrite, nitrosomorpholine, oxygen and nitric oxide were monitored during the experiments. RESULTS: The delivery of nitrite into HCl containing thiocyanate resulted in nitrosation of morpholine, with the rate of nitrosation being greatest at pH 2.5. Under anaerobic conditions, ascorbic acid converted the nitrite to nitric oxide and prevented nitrosation. However, in the presence of dissolved air, the ascorbic acid was ineffective at preventing nitrosation. This was due to the nitric oxide combining with oxygen to reform nitrite and this recycling of nitrite depleting the available ascorbic acid. Further studies indicated that the rate of consumption of ascorbic acid by nitrite added to natural human gastric juice (pH 1.5) was extremely rapid with 200 micromol/l nitrite consumed 500 micromol/l ascorbic acid within 10 s. CONCLUSIONS: The rapid consumption of ascorbic acid in acidic gastric juice by nitrite in swallowed saliva indicates that the potential for acid nitrosation will be maximal at the GO junction and cardia where nitrite first encounters acidic gastric juice. The high incidence of mutagenesis and neoplasia at this anatomical location may be due to acid nitrosation arising from dietary nitrate. PMID- 11916187 TI - Prevalence of subtypes of intestinal metaplasia in the general population and in patients with autoimmune chronic atrophic gastritis. AB - BACKGROUND: Gastric intestinal metaplasia (IM) is seen mostly in association with chronic gastritis, induced either by Helicobacter pylori infection or autoimmune mechanisms. IM can be categorized into three subtypes, where type III is associated with gastric carcinoma of intestinal type. METHODS: Gastric biopsies from 475 subjects randomly selected from the general population and from 27 patients with autoimmune gastritis associated with pernicious anaemia were used. The criteria of Filipe & Jass were applied using different histochemical techniques in combination with haematoxylin and eosin stained material. RESULTS: Twenty-three percent (109/475) of the subjects from the general population and 88% (24/27) in the group with autoimmune gastritis had IM. Type III IM occurred in 4% in both populations. Type III IM was located in the antrum in 90% in the general population. In the group with autoimmune gastritis, only one patient had type III IM, which was located in the corpus. CONCLUSIONS: This study reveals for the first time the prevalence and distribution of subtypes of IM in a general population from the Western world. The comparatively high prevalence of type III IM in the general population (4%) indicates that its role as a precursor of gastric carcinoma may have been overemphasized. A similar prevalence of type III IM in patients with autoimmune gastritis may be considered low and suggests that mechanisms for gastric carcinogenesis other than the atrophy-metaplasia-dysplasia sequence could also operate in this condition. PMID- 11916188 TI - Differential susceptibility of inbred mouse strains to Helicobacter pylori infection. AB - BACKGROUND: Host factors play an important part in the pathogenesis of Helicobacter pylori-associated disease. The aim of this study was to screen various inbred strains of mice for genetic differences in susceptibility to H. pylori infection. METHODS: Mice of strains BALB/cJ, C.B-17-Prkdc(scid), C3H/HeJ, C3H/HeN, C57BL/6J, C57BL/6J-I110(tm/Cgn), DBA/2J, and FVB/N were inoculated intragastrically with H. pylori SS1. At 1, 4 and 6 months after inoculation, mice were necropsied, and bacterial cultures and histologic studies of the stomachs were performed. RESULTS: Significant differences in the level of colonization by H. pylori were observed among inbred strains at each time of infection. These differences were most distinct at 4 months after inoculation with highest levels in strains C3H/HeJ and C3H/HeN and lowest in strains FVB/N, C57BL/6J and C57BL/6J I110(tm/Cgn). Infected mice revealed a mild increase in inflammatory cells compared with controls at 1 and 4 months, but not at 6 months after inoculation. The host strain effect on gastric disease was fairly mild, with two exceptions. Firstly, infected I110(tm/Cgn) mice developed a severe, hyperplastic gastritis, indicating that interleukin-10 is an important regulator of the inflammatory response to H. pylori. Secondly, infected C3H/HeN mice had a propensity to develop lymphoid aggregates in the gastric mucosa. CONCLUSIONS: The strain differences described here will be useful for the design of genetic mapping studies in mice to elucidate the genes controlling gastric infection by H. pylori. Our results further show that genetically altered mice are a valuable tool for identifying candidate genes possibly contributing to susceptibility to H. pylori infection. PMID- 11916189 TI - Gastrin and Helicobacter pylori in low-grade MALT lymphoma patients. AB - BACKGROUND: This study of patients with Helicobacter pylori infection and low grade MALT lymphoma aimed to investigate: 1) the effect of H. pylori eradication therapy on the serum gastrin level, 2) whether changes of the serum gastrin level after therapy could predict the prognosis of patients with this tumour, and 3) the relationship between the gastric H. pylori load, the serum gastrin level and the status of MALT lymphoma. METHODS: Thirteen patients with documented low-grade MALT lymphoma and H. pylori infection were enrolled and received H. pylori eradication therapy as the sole initial treatment. The presence of H. pylori, the serum gastrin level, the endoscopic findings, the pathologic features of the biopsies and resected specimens, and the endoscopic ultrasonography findings were evaluated before and after therapy. Follow-up was carried out every 3-6 months. RESULTS: H. pylori eradication was eventually achieved in all 13 patients. The pretreatment fasting serum gastrin level decreased from 177.1 +/- 107.4 pg/ml to 129.2 +/- 78.1, 96.4 +/- 66.6 and 80.1 +/- 42.7 pg/ml after 0-3, 3-6 and 6-9 months, respectively (all P < 0.05). Successful eradication of H. pylori was followed by a decrease of the fasting serum gastrin level and complete regression of initial low-grade MALT lymphoma was observed in all patients. However, two patients subsequently developed recurrent high-grade MALT lymphoma or high-grade lymphoma. In one of them, the serum gastrin level rose again above the pretreatment value. In the other, however, the fasting gastrin level fell throughout the study period. The median fasting serum gastrin level before H. pylori eradication therapy was higher in the patients with tumours of the gastric body (203.4 +/- 108.9 pg/ml) than in those with tumours of the antrum and angulus (89.3 +/- 28.0 pg/ml) (P = 0.06). CONCLUSIONS: Hypergastrinaemia may be associated with an increased risk of gastric MALT lymphoma. PMID- 11916190 TI - Role of anti-parietal cell antibody in Helicobacter pylori-associated atrophic gastritis: evaluation in a country of high prevalence of atrophic gastritis. AB - BACKGROUND: Helicobacter pylori plays an important part in the progression of atrophic gastritis; however, markers for predicting the progression of atrophic gastritis remain unidentified. We investigated the relation between the degree of atrophic gastritis and the amount of anti-parietal cell antibodies (APCAs) present. METHODS: In 219 Japanese patients, APCA was investigated by enzyme linked immunosorbent assay (ELISA) and by Western blotting. The grade of corpus atrophy was estimated by histology and serum pepsinogen levels. Serum levels of pepsinogen were evaluated by radioimmunoassay. RESULTS: Helicobacter pylori infection did not affect the APCA levels determined by ELISA. Long-term administration of proton-pump inhibitors and H. pylori eradication did not influence the levels of APCAs. However, in H. pylori-positive patients, the levels of APCA determined by ELISA were statistically higher in patients with severe atrophy than in those with mild atrophy as determined histologically (0.67+/-0.48 versus 0.45+/-0.40; A492, mean+/-s, P=0.01) and serologically by pepsinogen levels (0.66+/-0.51 versus 0.44+/-0.40. P=0.002). The levels of pepsinogen I/II ratio were correlated with APCA levels only in the H. pylori positive group. Western blotting showed that major antigen was identical with the beta-subunit of H+,K+-ATPase. CONCLUSION: APCA plays an important part in the progression of corpus atrophy after H. pylori infection. PMID- 11916191 TI - Are anxiety and depression related to gastrointestinal symptoms in the general population? AB - BACKGROUND: In clinical studies there is a strong relationship between gastrointestinal symptoms, anxiety and depression. The results may be biased, however, since anxiety and depression will influence the decision to consult a doctor. The aim of this study was to investigate the relationship between these symptoms in the population. METHODS: In the Health Study of Nord-Trondelag County of Norway (HUNT) a questionnaire concerning physical and mental health, demographic and life-style factors was sent to all inhabitants aged 20 years and above (a total of 94,197 persons). Valid questionnaires were returned by 62,651 persons (66.5%). Presence of nausea, heartburn, diarrhoea and constipation during the last year was self-reported. Anxiety disorders and depression were based on self-ratings of the Hospital Anxiety and Depression Scale (HADS). RESULTS: 48% of the population reported one or more of the four gastrointestinal symptoms. Based on the HADS ratings, 15.3% of the population had an anxiety disorder and 10.4% a depression. Anxiety disorder was most strongly associated with nausea (OR 3.42). Anxiety was also associated with heartburn, diarrhoea and constipation, but weaker than with nausea. Depression was less strongly associated with the four gastrointestinal symptoms. Demographic factors, life-style factors and extra gastrointestinal complaints could not explain the effect of anxiety disorders and depression on these gastrointestinal symptoms. CONCLUSIONS: In this population study there was a strong relationship between gastrointestinal symptoms, anxiety disorders and depression. These findings suggest that mental disorders in patients with gastrointestinal symptoms are not merely a consequence of selection bias in patient materials but connected to the symptoms themselves. PMID- 11916192 TI - Effects of calcium deficiency and calcium supplementation on gastrectomy-induced osteopenia in the young male rat. AB - BACKGROUND: Surgical removal of the stomach (gastrectomy, Gx) induces osteopenia. In this study we compared the osteopenic effect of Gx with that induced by calcium (Ca) deficiency. METHODS: Young male rats were subjected to Gx and/or low Ca diet (-Ca). A group of Gx rats received standard diet + oral Ca supplementation (+Ca). The rats were killed at various times after the operation/start of treatment (longest time 12 weeks). After 8 weeks on low Ca diet, the blood Ca2+ concentration was lowered slightly in both Sham-operated and Gx rats. The calvariae were subjected to transillumination analysis and quantitative histomorphometry. Also the tibiae were subjected to histomorphometry. RESULTS: Transillumination of the calvariae revealed extensive bone loss in the rats that had been subjected to Gx and/or low Ca diet. Gx + Ca induced the same bone loss as Gx alone. These observations were later confirmed in quantitative terms by histomorphometry (Sham-Ca 56%, Gx 35%, Gx + Ca 32%, Gx - Ca 58% less bone area than in Sham). The osteopenia induced by Gx + low Ca diet seetned more rapid in onset than that induced by Gx or low Ca diet alone. Tibiae from Gx rats and rats given a low Ca diet displayed a reduced trabecular bone volume (Sham-Ca 27% remaining, Gx 36%, Gx + Ca 44%, Gx - Ca 17%) and reduced trabecular number (Sham-Ca 44% remaining, Gx 41%, Gx + Ca 56%, Gx - Ca 33%). The trabecular thickness was reduced in the Gx rats and Gx - Ca rats (Gx 78% remaining, Gx - Ca 63%) but not in Sham-operated rats receiving a low Ca-diet (95% remaining). CONCLUSION: Although the pattern of osteopenia was qualitatively quite similar in Gx rats and Ca-deficient rats, in quantitative terms the low Ca diet was more detrimental to bone than Gx. Ca deficiency induced a similar degree of osteopenia in both Sham and Gx rats. Ca supplementation failed to prevent the Gx-induced osteopenia. PMID- 11916193 TI - Inflammatory bowel disease evaluated by low-field magnetic resonance imaging. Comparison with endoscopy, 99mTc-HMPAO leucocyte scintigraphy, conventional radiography and surgery. AB - BACKGROUND: To evaluate the use of low-field magnetic resonance imaging (MRI) in active inflammatory bowel disease (IBD). METHODS: MRI was executed in a consecutive cohort of 28 patients with Crohn disease (CD) and in 17 with ulcerative colitis (UC) prior to glucocorticoid treatment (1 mg prednisolone orally/kg body weight/day). MRI was repeated after 2-3 weeks (22 CD, 12 UC), and again after treatment completion or prior to surgery (18 CD, 6 UC). Five bowel segments were evaluated separately. MRIs were blindly evaluated by two observers, and findings compared with 39 leucocyte scintigraphies, 38 endoscopies, 15 double contrast barium enemas, 66 small-bowel radiographic examinations and surgery in 23 patients. RESULTS: In CD, blinded evaluation revealed a kappa (kappa) of 0.84 (95% confidence interval (CI) 0.78-0.91). In UC, kappa was 0.66 (95% CI 0.55 0.78). Agreements regarding disease extension between MRI and other modalities in CD were found in 345 bowel segments out of 391 (88.2%) at risk, and in UC in 209/235 (88.9%). Colonic disease activity gradings by radiography and endoscopy correlated significantly with T2-signal intensity (SI(T2)) and increments in T1 signal intensity (%SI(T1)) in both diseases. Significant correlations between MRI indices of disease activity and CDAI in CD (MRI-SI(T2): P <0.0001: MRI% SI(T1): P=0.0008) and the Powell-Tuck index in UC (MRI% SI(T1): P=0.008) were found. CONCLUSIONS: With low interobserver variation and high concordance of findings with other examinations. low-field MRI seems a valuable modality in active IBD. In addition, MRI expressions of disease activity correlate to clinical, radiographic and endoscopic disease activity. PMID- 11916194 TI - Interleukin-1beta and tumor necrosis factor-alpha induce chemokine and matrix metalloproteinase gene expression in human colonic subepithelial myofibroblasts. AB - BACKGROUND: Colonic subepithelial myofibroblasts may play a role in the inflammatory responses and in extracellular matrix (ECM) metabolism. In this study, we investigated the effects of interleukin (IL)-1beta and tumor necrosis factor (TNF)-alpha on chemokine (IL-8 and monocyte chemoattractant protein (MCP) 1) and ECM turnover (proliferation of subepithelial myofibroblasts, and secretion of ECM and matrix metalloproteinases (MMPs)) in colonic subepithelial myofibroblasts. METHODS: Human colonic subepithelial myofibroblasts were isolated using the method described by Mahida et al. Chemokine and MMP expressions were determined by ELISA and Northern blotting. Nuclear factor (NF)-kappaB and NF-IL6 DNA binding activities were evaluated by electrophoretic gel mobility shift assays (EMSA). RESULTS: IL-1beta and TNF-alpha did not affect the proliferation of subepithelial myofibroblasts, but stimulated the secretion of types I and IV collagens weakly. Unstimulated subepithelial myofibroblasts secreted a large amount of MMP-2, but a small amount of IL-8, MCP-1 and MMP-1. IL-1beta and TNF alpha both induced a dose- and time-dependent increase in IL-8, MCP-1 and MMP-1 secretion, and weakly stimulated MMP-2 secretion. IL-1beta and TNF-alpha both rapidly evoked NF-kappaB DNA-binding activity. The inhibition of NF-kappaB activation markedly blocked both IL-1beta- and TNF-alpha-induced IL-8 and MCP-1 mRNA expression, but did not affect MMP-1 mRNA expression. CONCLUSIONS: These observations indicate that chemokine secretion and ECM metabolism are collectively regulated by the proinflammatory cytokines, IL-1beta and TNF-alpha, in colonic subepithelial myofibroblasts. Thus, colonic subepithelial myofibroblasts may play an important role in the pathophysiology of inflammation in the colon. PMID- 11916195 TI - Effects of topical ropivacaine on eicosanoids and neurotransmitters in the rectum of patients with distal ulcerative colitis. AB - BACKGROUND: Topical administration of lidocaine has been suggested to have beneficial clinical effects in patients with active ulcerative colitis, but the mechanism of action, if any, remains obscure. As local anaesthetics may exert anti-inflammatory actions through their inhibition of nervous reflexes, we have studied the local effects of a single rectal dose of ropivacaine gel on rectal concentrations of eicosanoids and neurotransmittors in patients with relapsing ulcerative colitis. METHODS: In a randomized, double-blind, placebo-controlled study, concentrations of leukotriene B4, thromboxane B2 and prostaglandin E2 in rectal dialysates and concentrations of substance P, neurokinin A, somatostatin, vasoactive intestinal polypeptide and calcitonin gene-related peptide in rectal biopsies from 19 patients with active, distally located, ulcerative colitis were measured before and after rectal administration of a 200-mg dose of ropivacaine- or placebo-gel by use of radioimmunoassays. For comparison with normal conditions, concentrations of neuropeptides were measured in another 19 patients with relapsing ulcerative colitis and 14 controls with non-inflamed colon. RESULTS: No significant changes in concentrations of eicosanoids or neuropeptides were observed after ropivacaine or placebo administration. Baseline concentrations of all neuropeptides, except somatostatin, were significantly lower in active ulcerative colitis than in controls with non-inflamed colon. CONCLUSIONS: These findings reveal no evidence of anti-inflammatory actions by ropivacaine in active ulcerative colitis and thus provide no rationale for topical treatment with local anaesthetics. PMID- 11916196 TI - Mast cells involvement in the inflammation and fibrosis development of the TNBS induced rat model of colitis. AB - BACKGROUND: Mast cells have been implicated in chronic inflammatory conditions resulting in fibrosis, such as Crohn disease. However, a link between inflammation, fibrosis and mast cells has not been demonstrated in human or animal intestinal diseases. This work was undertaken to analyze whether mast cells play a role in inflammation and fibrosis in the TNBS-induced rat colitis. METHODS: Rats were rectally instilled 2,4,6,-trinitrobenzene sulfonic acid in ethanol, and immediately or 4 days later injected daily i.p. with nedocromil sodium, a mast cell stabilizer, compound 48/80, a mast cell activator, or saline. Rats were sacrificed 5 days post-TNBS, or on day 21. Intestinal inflammation and fibrosis were assessed by gross and histopathological evaluation. Colonic mast cell numbers (toluidine blue) and collagen (type I mRNA expression) were evaluated. Mast cell sonicate was added to rat colon fibroblasts. Fibroblast proliferation (3H-thymidine), collagen synthesis (3H-proline) and contractile activity (tridimensional collagen lattice contraction) were then assessed. RESULTS: Nedocromil reduced inflammation and fibrosis possibly by decreasing mast cell numbers and activation and consequent collagen production. Compound 48/80 slightly enhanced the severity of the disease by activating mast cells. Mast cells increased fibroblast proliferation, collagen production and contractile activity. CONCLUSIONS: Mast cells are involved in the gastrointestinal tract inflammation and fibrosis of the TNBS-colitis rats. PMID- 11916197 TI - Transjugular intrahepatic portosystemic shunting is not superior to endoscopic variceal band ligation for prevention of variceal rebleeding in cirrhotic patients: a randomized, controlled trial. AB - BACKGROUND: The aim of the present study was to compare the transjugular intrahepatic portosystemic shunt (TIPS) with variceal band ligation (VBL) in the prophylaxis of variceal rebleeding in patients with cirrhosis of the liver. METHODS: Fifty-four cirrhotic patients (21 Child-Pugh class A, 27 class B, 6 class C) were randomized to TIPS (n = 28) or VBL (n = 26) within 2 months after control of esophageal variceal hemorrhage. Statistical analysis was performed on the intention-to-treat principle. RESULTS: Mean follow-up was 2 years. Mortality risk at 1 and 2 years of follow-up was 7.8% +/- 5.3% and 19.9% +/- 8.8% in the TIPS group and 16.5% +/- 7.6% and 16.5% +/- 7.6% in the VBL group, respectively (n.s.); actuarial probability of remaining free from rebleeding was 83.7% +/- 77.4% and 71.4% +/- 10.4% in the TIPS group and 83.9% +/- 7.3% and 78.1% +/- 8.8% in the VBL group at 1 and 2 years, respectively (n.s.). Hepatic encephalopathy within 1 month after randomization was observed in 2 patients in the TIPS group and in 1 in the VBL group. CONCLUSION: TIPS is not superior to VBL in the prevention of variceal rebleeding. Furthermore, similar mortality rates in patients treated with TIPS or VBL negate TIPS as the preferred strategy for prevention of variceal rebleeding. PMID- 11916198 TI - Effect of interferon-alpha induction therapy on genotype 2b/3a and low viral load hepatitis C virus infection. A randomized multicentre study. AB - BACKGROUND: Interferon monotherapy for chronic hepatitis C virus (HCV) infection leads to sustained viral eradication in a minority of patients. However, in selected groups of patients, sustained virological response is observed in as many as 50% of patients. High initial interferon dose (induction therapy) has been reported to increase the initial response rate. We have studied the effect of interferon induction therapy in patients infected with HCV genotype 2b/3a, low viral load and no cirrhosis. METHODS: A total of 71 treatment-naive HCV RNA positive patients with biopsy-confirmed chronic hepatitis, with genotype 2b or 3a, viral load < or = 3 million copies per ml and no cirrhosis were randomized to receive either standard interferon therapy (3 MIU interferon-alpha-2a thrice weekly) for 26 weeks or 6 MIU interferon-alpha-2a daily for 4 weeks (induction group) followed by the standard dose (3 MIU thrice weekly) for 22 weeks. Those with persistent HCV RNA at 4 weeks stopped treatment. Patients were monitored for HCV RNA during and following treatment, and data were interpreted according to intention-to-treat analysis. RESULTS: Viral clearance occurred more rapidly (after 4 weeks) in the induction group (33/36 = 92%) compared to the standard interferon group (21/35 = 60%) (P = 0.01). Among the initial responders, 23/33 (induction group) compared to 16/21 (standard group) were persistently HCV RNA negative at the end of treatment. At 52 weeks (6 months' follow-up), 22/36 (61%) (induction group) compared to 10/35 (29%) (standard group) were HCV RNA-negative. Among initial responders, 22/33 (induction group) and 10/21 (standard group) achieved a sustained virological response. Among end-of-treatment responders, 22/24 (induction group) and 10/16 (standard group) were HCV RNA-negative at 6 months' follow-up (P = 0.013). CONCLUSIONS: In patients infected with HCV genotype 2b/3a, low viral load and without cirrhosis, IFN induction therapy increases the initial viral clearance and reduces the risk of relapse in end-of treatment responders. A sustained virological response was achieved in 61% of the patients receiving IFN induction therapy. PMID- 11916199 TI - Survival benefit of cirrhotic patients with hepatocellular carcinoma treated by percutaneous ethanol injection as a salvage therapy. AB - BACKGROUND: The therapeutic strategy for cirrhotic patients with hepatocellular carcinoma (HCC) who cannot tolerate surgery or transcatheter arterial chemoembolization (TACE) is uncertain. The safety and efficacy of percutaneous ethanol injection (PEI) as a salvage therapy in such patients is not clear. METHODS: A total of 63 (49 men) HCC patients (mean age 67 +/- 11 years), for whom surgery or TACE was not indicated because of the coexistence of various medical conditions, were enrolled and prospectively studied. Fifty-six (89%) were treated with PEI and 7 were treated with conservative measures. The outcome and the factors that may affect survival were evaluated. RESULTS: During a mean follow-up period of 16 +/- 9 months, 17 (30%) of the patients treated with PEI and 5 (71%) of those treated with conservative measures died (P = 0.045). A total of 16 patient-related and tumor-related variables that may influence the outcome were analyzed. Survival analysis showed that female gender, small (< or = 3 cm) solitary tumor and PEI were associated with a better prognosis (P < 0.05). When using the Cox proportional hazard model, PEI was the only significant independent factor predicting survival (relative risk: 0.3, 95% confidence interval: 0.11 0.86, P = 0.024). The 1- and 2-year survival rates were 85% and 65% for patients treated with PEI compared to 57% and 29% for conservative measures (P = 0.016). CONCLUSIONS: PEI may be a treatment option for cirrhotic patients who have HCC and coexisting contraindications that preclude surgery and TACE. Careful pre treatment patient selection may effectively prolong the survival. PMID- 11916200 TI - Alpha1-antitrypsin genotypes in patients with chronic pancreatitis. AB - BACKGROUND: An association between alpha1-antitrypsin deficiency and chronic pancreatitis (CP) has been reported in several case reports and two systematic studies. However, conflicting results have been shown in other studies of patients with CP. All previous studies were performed by phenotyping or by measurement of serum concentrations of alpha1-antitrypsin. The aim of this study was to investigate the relationship between alpha1-antitrypsin deficiency and CP by genetic analysis. METHODS: Ninety-six unrelated children and adolescents with idiopathic or hereditary CP and 185 healthy controls were enrolled. DNA was extracted from peripheral blood leukocytes and the exons 5 and 7 of the alpha1 antitrypsin gene were amplified by polymerase chain reaction using mutagenic forward primers introducing a Taq I restriction site. Genotyping of the S allele and the Z allele was performed by restriction fragment length polymorphism analysis using Taq I. RESULTS: Seven out of 96 patients (7.3%) with CP were heterozygous for an alpha1-antitrypsin deficiency allele (4 for the S allele and 3 for the Z allele). No patient was homozygous or compound heterozygous for these alleles. Twenty out of 185 control individuals (10.8%) were heterozygous for the S or Z allele (PiS: 12 controls; PiZ: 8 controls). No significant differences were found between the allele frequency in patients and the control individuals (P > 0.1). CONCLUSIONS: Alpha1-antitrypsin deficiency is not related to the pathogenesis of idiopathic or hereditary CP. PMID- 11916201 TI - Relevance of variants in serum antiproteinases for the course of chronic pancreatitis. AB - BACKGROUND: Mutations of the pancreatic serine protease inhibitor, Kazal type 1 (SPINK1), the cationic trypsinogen (PRSS1) and the cystic fibrosis transmembrane conductance regulator (CFTR) were reported to be genetic risk factors of chronic pancreatitis (CP). The aim of this study was to determine the role of genetic variants of the main serum antiproteinases alpha-1-antitrypsin (AAT) and alpha-2 macroglobulin (A2M) for the course of chronic pancreatitis. METHODS: 124 patients with non-alcoholic chronic pancreatitis (with PRSS1 or SPINK1 mutations or idiopathic pancreatitis) and 64 healthy controls were investigated for the AAT mutations PiS and PiZ, and the PiM determining variants R101H, V213A, E376D. In 101 subjects, the 'bait region' of A2M was sequenced. A pentanucleotide deletion in the bait region of A2M was analysed in 147 chronic pancreatitis (CP) patients and 87 controls. RESULTS: The lowest prevalences of V213A and E376D were found in PRSS1 patients, whereas an increased rate of these mutations was present in the SPINK1 group, and the highest prevalence was found in patients with idiopathic pancreatitis. The prevalence of PiM variants was higher in patients with early onset CP than in late onset (P < 0.05 for E376D). The coding region of the bait region of A2M was of wild type in all investigated subjects. The A2M pentanucleotide deletion showed a homogenous distribution in patients and controls. CONCLUSIONS: Our study suggests a moderating, but not predominant, role of AAT variants in the course of chronic non-alcoholic pancreatitis. PMID- 11916202 TI - Fatal hepatic failure after emergence of the hepatitis B virus mutant during lamivudine therapy in a patient with liver cirrhosis. AB - Lamivudine therapy for chronic hepatitis and decompensated liver cirrhosis related to the hepatitis B virus (HBV) resulted in improvement of liver function and inhibition of viral replication. Despite emergence of the HBV mutant, e antigen seroconversion and improvement of liver function may be achieved with continuation of lamivudine therapy. Although hepatic decompensation has been reported in a few cases after the emergence of lamivudine-resistant mutants, fatal cases of non-transplant patients have only rarely been reported in the literature. Here, we describe a patient with HBV-related liver cirrhosis who died after a breakthrough infection with a lamivudine-resistant mutant. Hepatic failure and mortality developed after flare-up of severe hepatitis after 13 months of lamivudine treatment. Emergence of the HBV mutant with substitution of isoleucine for leucine at residue 426 (L4261) in combination with isoleucine for methionine at residue 550 (M5501) was observed at 10 and 13 months of treatment. PMID- 11916203 TI - p53 protein expression in epithelial high-grade dysplasia from the stomach. PMID- 11916204 TI - Monitoring azathioprine metabolite levels and thiopurine methyl transferase (TPMT) activity in children with inflammatory bowel disease. PMID- 11916205 TI - The damaging effect of confirming feedback on the relation between eyewitness certainty and identification accuracy. AB - The authors investigated eyewitnesses' retrospective certainty (see G. L. Wells & A. L. Bradfield, 1999). The authors hypothesized that extemal influence from the lineup administrator would damage the certainty-accuracy relation by inflating the retrospective certainty of inaccurate eyewitnesses more than that of accurate eyewitnesses (N = 245). Two variables were manipulated: eyewitness accuracy (through the presence or absence of the culprit in the lineup) and feedback (confirming vs. control). Confirming feedback inflated retrospective certainty more for inaccurate eyewitnesses than for accurate eyewitnesses, significantly reducing the certainty-accuracy relation (from r = .58 in the control condition to r = .37 in the confirming feedback condition). Double-blind testing is recommended for lineups to prevent these external influences on eyewitnesses. PMID- 11916206 TI - The effects of jury size, evidence complexity, and note taking on jury process and performance in a civil trial. AB - A total of 567 jury-eligible men and women who were assigned to 6- or 12-person juries saw a videotaped civil trial that contained either I or 4 plaintiffs. Half the juries took notes, whereas the remainder did not. Six-person juries that did not take notes awarded multiple plaintiffs the highest amounts of compensation. Six-person juries also gave the highest punitive damages when they did not take notes and judged multiple plaintiffs. The punitive awards of 6-person juries were highly variable compared with 12-person juries. Multiple plaintiffs also increased the unpredictability of jury punitive awards. Twelve-person juries deliberated longer, recalled more probative information, and relied less than 6 person juries on evaluative statements and nonprobative evidence. Limitations and implications are discussed. PMID- 11916207 TI - Organizational citizenship behavior and workplace deviance: the role of affect and cognitions. AB - To investigate the role of affect and cognitions in predicting organizational citizenship behavior (OCB) and workplace deviance behavior (WDB), data were collected from 149 registered nurses and their coworkers. Job affect was associated more strongly than were job cognitions with OCB directed at individuals, whereas job cognitions correlated more strongly than did job affect with OCB directed at the organization. With respect to WDB, job cognitions played a more important role in prediction when job affect was represented by 2 general mood variables (positive and negative affect). When discrete emotions were used to represent job affect, however, job affect played as important a role as job cognition variables, strongly suggesting the importance of considering discrete emotions in job affect research. PMID- 11916208 TI - Assessing the construct validity of the job descriptive index: a review and meta analysis. AB - The construct validity of the Job Descriptive Index (JDI) was investigated by using a meta-analysis to summarize previous empirical studies that examined antecedents, correlates, and consequences of job satisfaction. In total, 79 unique correlates with a combined total of 1,863 correlations were associated with the JDI subdimensions. The construct validity of the JDI was supported by (a) acceptable estimates of internal consistency and test-retest reliability, (b) results that conform to a nomological network of job satisfaction relationships, and (c) demonstrated convergent and discriminant validity. Contrasting results with previous meta-analytic findings offered further support for the JDI's construct validity. Limitations of the JDI and suggestions for future research are discussed. PMID- 11916209 TI - Modifying supervisory practices to improve subunit safety: a leadership-based intervention model. AB - This article presents a leadership-based intervention model designed to modify supervisory monitoring and rewarding of subordinates' safety performance. Line supervisors received weekly feedback based on repeated episodic interviews with subordinates conceming the cumulative frequency of their safety-oriented interactions. This information identified the priority of safety over competing goals such as speed or schedules. Section managers received the same information and used it to communicate (high) safety priority. They also were trained to conduct episodic interviews to provide intermittent feedback after intervention, tuming safety priority into an explicit performance goal. Safety-oriented interaction increased significantly in the experimental groups but remained unchanged in the control groups. This change in safety-oriented interaction was accompanied by significant (and stable) changes in minor-injury rate, earplug use, and safety climate scores during the postintervention period. PMID- 11916210 TI - The interactive effects of conscientiousness and agreeableness on job performance. AB - The authors hypothesized that the relationship between conscientiousness and job performance would be stronger for persons high in agreeableness than for those low in agreeableness. Results of hierarchical moderated regression analyses for 7 independent samples of employees across diverse occupations provided support for the hypothesis in 5 of the samples. In samples supporting the hypothesis, among the highly conscientious workers, those low in agreeableness were found to receive lower ratings of job performance than workers high in agreeableness. One explanation for lack of an interaction between conscientiousness and agreeableness in the other 2 samples is that those jobs were not characterized by frequent, cooperative interactions with others. Overall, the results show that highly conscientious workers who lack interpersonal sensitivity may be ineffective, particularly in jobs requiring cooperative interchange with others. PMID- 11916211 TI - Why are structured interviews so rarely used in personnel selection? AB - By adopting the theory of planned behavior, this study tried to predict human resources managers' (N = 79) intentions toward unstructured and structured interview techniques. Managers evaluated case descriptions of both techniques and were interviewed about their own practices. The data revealed stronger intentions toward unstructured interviewing than toward structured interviewing, which was consistent with their own practices in selecting staff, which appeared to be rather unstructured. l. Ajzen's (1991) theory appeared to be a useful framework for predicting managers' intentions. In particular, attitudes and subjective norms were predictive of intentions to engage in either method. Only intentions toward the unstructured case were related to managers' actual behavior. PMID- 11916212 TI - Effects of procedure and outcome accountability on interview validity. AB - This study tested effects of holding interviewers accountable for either the procedure they follow to make interview judgments (procedure accountability) or the accuracy of their judgments (outcome accountability) on interview validity. Undergraduates (N = 338) simulated employment interviewers in an experiment that crossed 2 levels of procedure accountability with 2 levels of outcome accountability. They watched videotapes of 60 managers answering an interview question and rated the managers on leadership potential. The dependent variable was the correlation between their interview judgments and supervisory ratings of the managers' actual leadership performance on the job. Results showed that procedure accountability increased interview validity and outcome accountability lowered it. Participants' apparent attentiveness fully mediated effects of procedure accountability on interview validity. PMID- 11916213 TI - The impact of cross-training on team effectiveness. AB - The authors examined the role of cross-training in developing shared team interaction mental models, coordination, and performance in a 2-experiment study using computer simulation methodology (for Experiment 1, N = 45 teams; for Experiment 2, N = 49 teams). Similar findings emerged across the 2 experiments. First, cross-training enhanced the development of shared team-interaction models. Second, coordination mediated the relationship between shared mental models and team performance. However, there was some inconsistency in the findings concerning the depth of cross-training necessary for improving shared mental models. Results are discussed in terms of the impact of different levels of cross training on team effectiveness. PMID- 11916215 TI - Personality and job performance: test of the mediating effects of motivation among sales representatives. AB - Research shows consistent relations between personality and job performance. In this study the authors develop and test a model of job performance that examines the mediating effects of cognitive-motivational work orientations on the relationships between personality traits and performance in a sales job (N = 164). Covariance structural analyses revealed proximal motivational variables to be influential mechanisms through which distal personality traits affect job performance. Specifically, striving for status and accomplishment mediate the effects of Extraversion and Conscientiousness on ratings of sales performance. Although Agreeableness was related to striving for communion, neither Agreeableness nor communion striving was related to success in this sales job. The importance of the proposed motivational orientations model is discussed. PMID- 11916214 TI - From acclaim to blame: evidence of a person sensitivity decision bias. AB - In a series of studies, the authors established empirical support for a general decision-making bias that they termed a person sensitvity bias. Specifically, a person sensitivity bias consists of a person positivity bias (D. O. Sears, 1983) under positive performance conditions and a person negativity bias under negative performance conditions. The authors conducted the first empirical studies of a direct comparison between individuals and objects performing the same task under both positive and negative performance conditions. Two additional studies tested the boundaries of the sensitivity bias within negatively framed decision dilemmas. The results are discussed in terms of their relevance toward a more comprehensive theory of person-object evaluation differences. PMID- 11916216 TI - The nature and dimensionality of organizational citizenship behavior: a critical review and meta-analysis. AB - This article reviews the literature on organizational citizenship behavior (OCB) and its dimensions as proposed by D. W. Organ (1988) and other scholars. Although it is assumed that the behavioral dimensions of OCB are distinct from one another, past research has not assessed this assumption beyond factor analysis. Using meta-analysis, the authors demonstrate that there are strong relationships among most of the dimensions and that the dimensions have equivalent relationships with the predictors (job satisfaction, organizational commitment, fairness, trait conscientiousness, and leader support) most often considered by OCB scholars. Implications of these results are discussed with respect to how the OCB construct should be conceptualized and measured in the future. PMID- 11916217 TI - The relative importance of task, citizenship, and counterproductive performance to global ratings of job performance: a policy-capturing approach. AB - A review of research on job performance suggests 3 broad components: task, citizenship, and counterproductive performance. This study examined the relative importance of each component to ratings of overall performance by using an experimental policy-capturing design. Managers in 5 jobs read hypothetical profiles describing employees' task, citizenship, and counterproductive performance and provided global ratings of performance. Within-subjects regression analyses indicated that the weights given to the 3 performance components varied across raters. Hierarchical cluster analyses indicated that raters' policies could be grouped into 3 homogeneous clusters: (a) task performance weighted highest, (b) counterproductive performance weighted highest, and (c) equal and large weights given to task and counterproductive performance. Hierarchical linear modeling indicated that demographic variables were not related to raters' weights. PMID- 11916218 TI - Disentangling the effects of voice: the incremental roles of opportunity, behavior, and instrumentality in predicting procedural fairness. AB - Many voice studies have failed to distinguish among voice opportunity, perceived voice opportunity, voice behavior, and voice instrumentality. Thus, the authors sought to clarify the roles of each in determining procedural fairness perceptions. Controlling for the effect of voice opportunity, each of the 3 remaining constructs was hypothesized to predict fairness. Furthermore, voice instrumentality was hypothesized to moderate the effect of voice behavior on fairness. Undergraduates (N = 102; 81 for some analyses) participated in an orientation-week design simulation in which voice opportunity was manipulated. The results indicated significant incremental effects of perceived voice opportunity and the predicted Voice Instrumentality x Voice Behavior interaction. Fairness was lowest for individuals who were denied voice opportunity, perceived less voice opportunity, and provided high levels of noninstrumental voice behavior. PMID- 11916219 TI - Cognitions and behavior in asymmetric social dilemmas: a comparison of two cultures. AB - This study investigated whether cognitions and behavior in an asymmetric social dilemma can be predicted by national culture. Results indicated that, as predicted, groups of decision makers from Japan--a collectivist, hierarchical culture-were more cooperative, expected others to be more cooperative, and were more likely to adopt an equal allocation distribution rule to resolve the dilemma than were groups of decision makers from the United States-an individualist, egalitarian culture. An opportunity for communication had a greater impact on expectations of others' behavior in groups of U.S. decision makers than in groups of Japanese decision makers. PMID- 11916220 TI - Comparing meta-analytic moderator estimation techniques under realistic conditions. AB - One of the most problematic issues in contemporary meta-analysis is the estimation and interpretation of moderating effects. Monte Carlo analyses are developed in this article that compare bivariate correlations, ordinary least squares and weighted least squares (WLS) multiple regression, and hierarchical subgroup (HS) analysis for assessing the influence of continuous moderators under conditions of multicollinearity and skewed distribution of study sample sizes (heteroscedasticity). The results show that only WLS is largely unaffected by multicollinearity and heteroscedasticity, whereas the other techniques are substantially weakened. Of note, HS, one of the most popular methods, typically provides the most inaccurate results, whereas WLS, one of the least popular methods, typically provides the most accurate results. PMID- 11916221 TI - Proceedings of the First International Conference on Recent Advances and Future Directions in Endocrine Therapy for Breast Cancer: summary consensus statement. PMID- 11916222 TI - Estrogen receptor: current understanding of its activation and modulation. AB - Breast cancer development and progression are directly related to the effects of the female hormone estrogen. The nuclear receptor for estrogen (ER) functions as a transcription factor controlling estrogen-regulated genes. Receptor conformation on ligand binding, its interaction with various coregulators, and response elements in the promoter region of target genes all contribute to the net estrogenic effects in a cell. ER is an important diagnostic and therapeutic target in breast cancer. Various polypeptide growth factors and their membrane receptors also contribute to breast cancer development and progression. Pathways mediating cell survival, cell proliferation, and response to stress not only generate signals through various protein kinase pathways to enhance cell survival and proliferation, but these pathways also interact with ERs. Kinases in the growth factor cascade can phosphorylate and activate ER, and ER in turn activates and augments signaling through the growth factor pathways. Signaling through the growth factor pathways may contribute to hormonal resistance states by ligand independent activation of ER. Targeting growth factor pathways, in addition to ER, is a developing strategy that hypothetically may represent optimal therapy by preventing the development of resistance to endocrine therapy. PMID- 11916223 TI - Aromatase inhibition and inactivation. AB - Aromatase is the key enzyme in the synthesis of estrogens and mediates the conversion of androstenedione and testosterone to estrone and estradiol. Because of the importance of estrogen in stimulating breast cancers, the inhibition of estrogen synthesis is a logical approach to treatment. Aromatase is an excellent target for inhibition, because it is the last step in steroid biosynthesis, and, therefore, there are no important downstream enzymes to be affected. In addition, although aromatase is a P-450 enzyme and shares common features with other enzymes in this class, such as liver metabolizing enzymes and steroidogenic enzymes, it has unique features in the aromatizing reaction, features that are amenable to the development of selective inhibition. The approach we took to develop the first aromatase inhibitors was to design substrate analogues based on the structure of androstenedione. Some of these inhibitors, such as 4 hydroxyandrostenedione [4-OHA (later known as formestane)], also cause enzyme inactivation. Instead of being released at the end of the reaction, the substrate analogue remains bound. Therefore, the inhibitor is not required to be present at all times to maintain inhibition, and it has high enzyme specificity. Subsequently, other investigators have taken a different approach to developing compounds based on inhibitors of P-450 enzymes. High selectivity has been achieved with some of these reversible inhibitors. We have developed a unique animal model with human tumors to compare the antitumor efficacy of antiestrogens and aromatase inhibitors and to optimize their use in sequence and combination as a guide for future clinical trials. PMID- 11916224 TI - Prospects for combining hormonal and nonhormonal growth factor inhibition. AB - In patients with estrogen receptor (ER)-negative disease or ER+ hormone-resistant disease, the dominant influence on tumor cell growth is growth factors, e.g., epidermal growth factor (EGF), heregulins, and insulin-like growth factors acting through specific receptor tyrosine kinases at the cell surface. This superfamily of ligand-activated growth factor receptors triggers cascades of biochemical signals that influence tumor cell motility, invasiveness, angiogenesis, and survival, as well as proliferation. In breast tumors, expression of epidermal growth factor receptor (EGFR) and/or erbB2 is associated with poor prognosis; the therapeutic utility of blocking these receptors has been established using trastuzumab (Herceptin), a monoclonal antibody that blocks erbB2 signaling. An alternative therapeutic approach is offered by small molecule inhibitors of EGFR TK, exemplified by ZD1839 (Iressa), a potent and selective EGFR-TK inhibitor. Resistance to tamoxifen is associated with up-regulation of the EGFR-TK pathway and mitogen-activated protein kinase activity is substantially increased in tamoxifen-resistant MCF-7 cells. ZD1839 treatment of tamoxifen-resistant MCF-7 cells blocks mitogen-activated protein kinase activity. Furthermore, treatment of wild-type MCF-7 cells with tamoxifen and ZD1839 prevents development of tamoxifen resistance. These data support the potential clinical utility of ZD1839 in tamoxifen-resistant breast cancer and suggest the possibility of preventing resistance by the early use of combination ZD1839 with antiestrogenic agents such as tamoxifen or ICI 182,780. PMID- 11916225 TI - The role of tamoxifen and aromatase inhibitors/inactivators in postmenopausal patients. AB - The traditional hormonal cascade of the 1970s and 1980s used tamoxifen followed by megestrol acetate and subsequently by aminoglutethimide. In the 1990s, however, three trials of third-generation aromatase inhibitors (AIs) compared with megestrol acetate and two trials of third-generation AIs compared with aminoglutethimide showed improved efficacy and decreased toxicity for the newer AIs. Thus, the hormonal cascade changed in the late 1990s, to one in which tamoxifen, followed by a third-generation AI, followed by megestrol acetate, seemed more suitable. Now, however, several trials comparing anastrozole, letrozole, and exemestane to tamoxifen as first-line hormonal agents for metastatic breast cancer have shown that these drugs are at least equivalent and perhaps superior to tamoxifen in that setting in terms of response rate and time to progression. Results from 1021 patients randomized to receive anastrozole versus tamoxifen showed a slightly improved overall response rate (RR; 29% versus 26%), slightly improved clinical benefit (CB; 57% versus 52%), and a significantly improved time to progression (TTP; 8.5 months versus 7.0 months) in favor of anastrozole. In 907 women randomized to treatment with letrozole versus tamoxifen, significantly improved RR (30% versus 20%), CB (49% versus 38%), and TTP (9.4 months versus 6 months) have all been shown for those treated with letrozole. In addition, a randomized Phase II trial of 121 patients has shown nonsignificant benefits in favor of exemestane (RR 41% versus 14%; CB 56% versus 42%; TTP not available). To date, none of these trials has demonstrated any overall survival benefit. Additional follow-up in regard to survival in the trial of tamoxifen versus letrozole and an expanded Phase III trial of tamoxifen versus exemestane are ongoing. PMID- 11916226 TI - Are differences in the available aromatase inhibitors and inactivators significant? AB - Aromatase inhibitors are endocrine agents with a different mode of action than tamoxifen against breast tumors. In postmenopausal women, estrogen concentrations are maintained primarily via aromatase, a cytochrome P-450 enzyme that acts at the final step in the estrogen synthesis pathway. The first clinically available aromatase inhibitor, aminoglutethimide, was introduced for the second-line treatment of advanced breast cancer in the late 1970s. Despite proven efficacy in this setting, its widespread use was limited by its overall toxicity and its lack of selectivity for the aromatase enzyme. This led to the search for novel, more effective, and less toxic aromatase inhibitors. As a result, several aromatase inhibitors with a high degree of selectivity for aromatase and improved tolerability have become clinically available for the treatment of postmenopausal women with advanced breast cancer: (a) anastrozole; (b) letrozole; (c) fadrozole; (d) formestane; and (e) exemestane. Of these, formestane and exemestane are steroidal nonreversible aromatase inhibitors, also known as aromatase inactivators, whereas fadrozole, anastrozole, and letrozole are nonsteroidal reversible aromatase inhibitors. These agents differ in pharmacokinetics, selectivity, and potency, although all are more selective than aminoglutethimide. Some differences in adverse effect profile are also noticeable between and within these two classes of agents. The clinical significance of these differences is not yet evident but may well prove to be relevant in the long-term adjunctive setting. PMID- 11916227 TI - Preliminary experience with pure antiestrogens. AB - Endocrine therapy plays an important role in the management of all stages of breast cancer, and the development of effective endocrine therapies has focused on modifying the response of hormone-sensitive tumors to estrogen. The most widely used agents are the nonsteroidal antiestrogens or selective estrogen receptor (ER) modulators, the class to which tamoxifen, the standard in terms of antiestrogen therapy, belongs. Tamoxifen is effective in both adjuvant and first line settings and is now used as prophylactic therapy in high-risk individuals. However, the partial agonist activity of tamoxifen on the uterus, coupled with "tumor flare" and the development of tamoxifen resistance, has limited its therapeutic utility. Attempts to eliminate the partial agonist activity of tamoxifen and increase its potency have led to the development of fulvestrant (Faslodex), the first in a new class of pure antiestrogens, the ER down regulators. Fulvestrant has a high affinity for the ER compared with tamoxifen but has none of the agonist activities of tamoxifen. This new agent is showing promising clinical activity in the treatment of advanced breast cancer after tamoxifen therapy. Fulvestrant's lack of ER agonist activity may provide a longer duration of response compared with tamoxifen, as it did in a preclinical setting. Fulvestrant has demonstrated that it is at least as effective as the third generation aromatase inhibitor anastrozole in patients whose disease has relapsed or recurred on prior endocrine therapy and is currently being evaluated in Phase III trials versus tamoxifen for the first-line therapy of advanced breast cancer. Future clinical studies will evaluate fulvestrant in adjuvant and neoadjuvant settings, together with its optimal sequencing in relation to tamoxifen and other endocrine therapies. PMID- 11916228 TI - Endocrine manipulation in advanced breast cancer: recent advances with SERM therapies. AB - Tamoxifen is one of the most effective treatments for breast cancer through its ability to antagonize estrogen-dependent growth by binding estrogen receptors (ERs) and inhibiting breast epithelial cell proliferation. However, tamoxifen has estrogenic agonist effects in other tissues such as bone and endometrium because of liganded ER-activating target genes in these different cell types. Several novel antiestrogen compounds have been developed that are also selective ER modulators (SERMs) but that have a reduced agonist profile on breast and gynecological tissues. These SERMs offer the potential for enhanced efficacy and reduced toxicity compared with tamoxifen. In advanced breast cancer clinical data exist for three first-generation SERMs (toremifene, droloxifene, idoxifene), which are related to the triphenylethylene structure of tamoxifen. In Phase II trials in a total of 263 patients resistant to tamoxifen, the median objective response rate to these SERMs was only 5% (range, 0-15%), with stable disease for > or =6 months in an additional 18% (range, 9-23%). As first-line therapy for advanced breast cancer, the median response rate was 31% (range, 20-68%) with a median time to progression of 7 months. Randomized Phase III trials for toremifene and idoxifene in more than 1500 patients showed no significant difference compared with tamoxifen. Fewer clinical data exist for the structurally distinct second- and third-generation SERMs (raloxifene, arzoxifene, EM-800, and ERA-923), although a similarly low median response rate of 6% (range, 0-14%) was seen in Phase II trials in tamoxifen-resistant patients. It remains unclear whether any clinical advantage exists for second- and third-generation SERMs over tamoxifen as first-line therapy. With the emergence of potent aromatase inhibitors (AIs) that are superior to tamoxifen, the clinical questions in advanced disease have shifted to which antiestrogen (including SERMs) may be effective following failure of AIs, and whether any merit exists for combined AI/SERM therapy. The main advantage for SERM therapy probably remains in early stage-disease (adjuvant therapy or prevention), in which the estrogenic effects on bone and reduced gynecological side effects may prove more beneficial than either tamoxifen or AI. The issue is whether the current clinical data for SERMs in advanced breast cancer are sufficiently strong to encourage that further development. PMID- 11916229 TI - Neoadjuvant endocrine therapy for breast cancer: medical perspectives. AB - The indolent nature of estrogen-dependent breast cancer is the most important obstacle for development of new adjuvant endocrine treatments. Clinical trials require thousands of study participants and at least a decade of clinical investigation. How can we be sure that a new endocrine agent warrants this extraordinary level of investment? Traditionally, we have relied on advanced breast cancer trials to determine which drugs are suitable for adjuvant studies. However, with endocrine agents the high incidence of resistance in metastatic breast cancer may mask important advances in efficacy. Recent clinical results with the aromatase inhibitor letrozole suggest that neoadjuvant endocrine therapy is a highly informative additional approach to consider when planning adjuvant studies. In this report, new neoadjuvant endocrine therapy study designs are discussed that address the following issues: (a) the scientific opportunities afforded by gene microarray studies and other genetic technologies to investigate the molecular basis of estrogen-dependent breast cancer; (b) studies that address critical drug development questions as a prelude to adjuvant studies; and (c) the conduct of randomized trials that compare neoadjuvant chemotherapy with neoadjuvant aromatase inhibitor therapy to establish a place for neoadjuvant endocrine therapy in routine clinical practice. PMID- 11916230 TI - Current status of adjuvant endocrine therapy for breast cancer. AB - Adjuvant endocrine therapy is a vitally important modality for treatment of women with resected hormone-sensitive breast cancer as indicated by the presence of estrogen and/or progesterone receptors in the tumor. It is not recommended in those patients whose tumors lack these hormone receptors. Tamoxifen for 5 years has been shown to reduce the risk of recurrence and death irrespective of menopausal status or use of adjuvant chemotherapy. In postmenopausal women, the major research emphasis relates to the use of third-generation aromatase inhibitors in sequence with, instead of, and in combination with tamoxifen. In premenopausal women, ovarian ablation/suppression has demonstrated efficacy comparable with some chemotherapy regimens, but its role in women receiving chemotherapy remains to be determined. PMID- 11916232 TI - Future use of selective estrogen receptor modulators and aromatase inhibitors. AB - Selective estrogen receptor modulators (SERMs) may act as estrogens or antiestrogens depending on the cell and tissue targets. The triphenylethylene SERMs are represented by tamoxifen and toremifene and a new agent with a novel carboxylic acid side chain (GW5638). Because of isomerization in the triphenylethylene molecule, "fixed ring" SERMs were introduced. The major one in development is the benzothiophene arzoxiphene (LY353381), which is now in Phase III clinical trial versus tamoxifen. A fourth group of SERMs is based on the estrogen molecule and comprises the so-called "pure" antiestrogen ICI 182,780 (fulvestrant, Faslodex) and a new oral analogue just entering trials, SR16234. The steroidal aromatase inhibitors [AIs (40H androstenedione 'and exemestane)] inactivate aromatase, whereas the triazole AIs (anastrozole and letrozole) inhibit the enzyme via the heme prosthetic group. Thus, there are two groups of AIs that show relative non-cross-resistance in advanced breast cancer and four groups of SERMs that also show a high degree of non-cross-resistance. With six different treatments and six or more clinical situations (prevention, neoadjuvant, adjuvant, and first- and second-line treatments for advanced disease) in which they may be used, the possible combinations of treatment are enormous. At present, we have few clinical pointers to optimal sequence of treatments. Now that most of the appropriate comparative trials have been performed, it may be the time to initiate novel approaches. These include alternating and sequential treatments, preferably with treatments changed before overt progression occurs. PMID- 11916231 TI - Preliminary data from ongoing adjuvant aromatase inhibitor trials. AB - With recent results showing letrozole and anastrozole to be superior to tamoxifen as initial therapy for advanced disease, the aromatase inhibitors are poised to establish their place in the adjuvant therapy of postmenopausal receptor-positive breast cancer. A review of the rationale, design, and preliminary results of the ongoing adjuvant trials that include aromatase inhibitors will be presented, along with the ongoing or planned substudies. Two strategies employing aromatase inhibitors after tamoxifen are being evaluated. The MA.17 international intergroup trial is randomizing postmenopausal patients who are disease-free after 5 years of adjuvant tamoxifen to an additional 5 years of letrozole or placebo. In a similar design, the National Surgical Adjuvant Breast and Bowel Project (NSABP) B33 trial is randomizing this patient population to 2 years of exemestane or placebo after the standard 5 years of adjuvant tamoxifen. The second approach under study is the use of both aromatase inhibitor and tamoxifen in sequence within the first 5 postoperative years. The International Cancer Collaboration Group (ICCG) trial is comparing 2 years of exemestane after 3 years of tamoxifen to a standard 5-year course of tamoxifen. Similarly, the ARNO trial is comparing 5 years of tamoxifen versus 2 years of tamoxifen followed by 3 years of anastrozole. In a four-arm study Breast International Group/Femara-Tamoxifen (BIG/FEMTA) conducted by the BIG, one arm contains letrozole given for 3 years after 2 years of tamoxifen. Several trials are investigating the role of anastrozole, letrozole, or exemestane as a 5-year adjuvant therapy to replace the standard 5 years of tamoxifen. Only the Arimidex and Tamoxifen, Alone or in Combination (ATAC) trial is testing a 5-year combination of tamoxifen plus an aromatase inhibitor in this setting. Companion studies of effects on end-organs other than the breast are ongoing in a number of these trials. Aromatase inhibitors are poised to alter the treatment paradigm of breast cancer and hopefully improve outcome for a substantial number of patients. PMID- 11916233 TI - Follow-up of the breast cancer prevention trial and the future of breast cancer prevention efforts. AB - Women who are at increased risk for developing breast cancer can be identified using quantitative risk assessment models that provide valid estimates of risk. The Breast Cancer Prevention Trial (BCPT, P-1) demonstrated that tamoxifen can reduce the incidence of both invasive and noninvasive breast cancer as well as of bone fractures in women at increased risk. These benefits accrue at the expense of increased risk of endometrial cancer, thromboses, cataracts, and possibly diminished quality of life in postmenopausal women. All premenopausal women with a 5-year risk of developing invasive breast cancer greater than 1.67% derive net benefit from using tamoxifen to reduce the risk. Subset analyses of older postmenopausal women taking raloxifene for the treatment of osteoporosis indicate reduction of breast cancer incidence by more than 70%. These findings led the National Surgical Adjuvant Breast and Bowel Project (NSABP) to design and launch the STAR trial (P-2, the Study of Tamoxifen and Raloxifene). Eligible women are at least 35 years of age and postmenopausal, and they must have either lobular carcinoma in situ (LCIS) or a 5-year risk of invasive breast cancer of at least 1.67% as determined by the Gail model [M. H. Gail et al., J. Natl. Cancer Inst. (Bethesda), 81: 1879-1886, 1989]. Subjects are randomly assigned to receive either tamoxifen 20 mg or raloxifene 60 mg daily in a double-blind, double-dummy design. The trial is designed to recruit a total of 22,000 postmenopausal women and is powered to demonstrate superior efficacy of either agent or their equivalence in reducing the incidence of primary breast cancer. Additional end points will include the incidence of cardiovascular events and bone fractures. Thromboembolic events and endometrial cancer are the predicted toxicities. Ancillary studies of cognitive function will also be performed. Raloxifene should not be used for the reduction of breast cancer risk outside the context of the STAR trial. PMID- 11916234 TI - Benign breast disease and breast cancer risk: potential role for antiestrogens. AB - Evidence from clinical follow-up studies has indicated that there is a relationship between the presence of histologically proven benign breast disease and breast cancer risk, and that the risk varies according to the histological category of benign breast disease and hormonal status. The risk associated with these histological factors appears to be equal in both breasts, suggesting that these factors are best considered markers of generalized increased breast cancer risk rather than direct precursor lesions. A number of interesting observations gleaned from the Nashville Study, the Breast Cancer Detection Demonstration Project and the Nurses' Health Study, among other studies, provide evidence of an interaction between these histological risk factors and estrogen in determining the level of breast cancer risk. In the National Surgical Adjuvant Breast Project P-1 trial, tamoxifen was associated with an 86% reduction in breast cancer risk among a small subset of women with biopsy-proven atypical hyperplasia. Taken together, these observations strongly suggest that there is an interaction between estrogen and histological factors in determining breast cancer risk, and that it may be possible to reduce the risk associated with these histological risk factors using antiestrogen therapy. PMID- 11916235 TI - The potential for aromatase inhibition in breast cancer prevention. AB - Although SERMs are currently being evaluated and are approved for breast cancer prevention in several countries, aromatase inhibitors and inactivators may represent interesting options in this setting. The encouraging results revealing these drugs to be superior to conventional therapy in metastatic breast cancer confirm their therapy efficacy and suggest that they may also have a role in adjuvant therapy and even for breast cancer prevention. Secondly, whereas the bulk of "high-risk" breast cancer patients with confirmed founder mutations in the BRCA1 or BRCA2 genes develop their cancers earlier in life (during the premenopausal period), 75-80% of all breast cancers, in general, develop in postmenopausal women. Thus, in considering prevention of breast cancer in moderate-risk groups, strategies for prevention in postmenopausal women may play an important role. Also, among high-risk patients who have not developed breast cancer by the time of the menopause, aromatase inhibition could be a feasible option. Considering the potential hazards of long-term use of SERMs, switching to an aromatase inhibitor or inactivator in this setting may be beneficial. Finally, the observation that postmenopausal estrogen levels are related to subsequent risk of breast cancer in the general population underlines the potential for estrogen suppression as a preventive strategy. Results from ongoing studies examining the toxicity of aromatase inhibitors and inactivators in postmenopausal women will set the stage for future trials that explore them as preventive treatment options. PMID- 11916236 TI - Cross-talk among estrogen receptor, epidermal growth factor, and insulin-like growth factor signaling in breast cancer. AB - Since the cloning of the estrogen receptor alpha (ERalpha) and subsequent identification of a second distinct form of ER, termed ERbeta, a large volume of research has begun to define the molecular mechanisms of ER action. However, although great progress has been made, ER action is still poorly understood. It is expected that a better understanding of ER action may lead to novel strategies and targets for breast cancer prevention and treatment. One of the early-realized functions of the ER was regulation of growth factor signaling, but the degree of interaction between these two mitogenic signaling pathways could not have been anticipated. Recent evidence suggests that the ER and the growth-factor-signaling pathways intersect and directly interact at every level of signal transduction. The resulting synergism between ER and growth factors has been documented both in normal breast development and, importantly, in breast cancer progression and antiestrogen resistance. In this review, we will highlight our current understanding of the molecular mechanisms of cross-talk between ER and growth factor-signaling pathways. PMID- 11916237 TI - Inhibition of erbB receptor (HER) tyrosine kinases as a strategy to abrogate antiestrogen resistance in human breast cancer. AB - It has been proposed that binding of ligand to the estrogen receptor (ER) releases its association with transcriptional corepressors, allowing the ER to recruit coactivators, which possess histone acetylase activity, and induce transcription of gene promoters containing estrogen response elements. It has also been proposed that the antiestrogen tamoxifen recruits transcriptional corepressors to the AF-2 region of the hormone-binding domain of the ER, thus blocking ER-mediated transcription. The ER cross-talks with a number of mitogenic signaling pathways and second messengers, like the epidermal growth factor receptor, the insulin-like growth factor-I receptor, mitogen-activated protein (MAP) kinase, phosphatidylinositol-3 kinase/Akt, dopamine, and cyclic AMP. Some of these molecules may: (a) support ligand-independent ER transcription; (b) increase the association of ER with coactivators of transcription; and/or (c) reduce the antiestrogen-induced association of ER with corepressors. These events either alone or in combination may result in hormone independence and/or antiestrogen resistance. We have examined whether signaling by HER2/neu (erbB-2) receptor tyrosine kinase, which can induce antiestrogen resistance, can also disrupt the tamoxifen-induced interaction of ER with transcriptional corepressors. Notably, tamoxifen-induced association of ER with the transcriptional corepressors N-CoR or SMRT was reduced in HER2-overexpressing breast tumor cells but not in cells with low HER2 levels. Small molecule inhibitors of the HER2 kinase or MAP extracellular signal-regulated kinase 1/2 or dominant-negative MAP extracellular signal-regulated kinase 1/2 constructs restored the inhibitory effect of tamoxifen on both ER-mediated transcription and tumor cell proliferation. Treatment with both tamoxifen and the small molecule HER1/2 kinase inhibitor AG1478 reduced mitogen-activated protein kinase activity and markedly reduced growth of established MCF-7/HER2 xenografts in athymic nude mice. Similar results have been obtained with ZD1839 ("Iressa"), an epidermal growth factor receptor (HER1) tyrosine kinase inhibitor. Taken together, these data suggest that exogenous inhibitors of the HER-signaling network and other mitogenic pathways can abrogate or delay the emergence of antiestrogen resistance, thus providing an evaluable therapeutic strategy in human breast carcinoma. PMID- 11916238 TI - Antitumor effects on human melanoma xenografts of an amplicon vector transducing the herpes thymidine kinase gene followed by ganciclovir. AB - Herpes simplex virus type-1 (HSV-1) has been demonstrated as a potentially useful gene delivery vector for gene therapy due to its high efficiency of in vivo transduction. The helper virus-dependent, HSV- 1 amplicon vectors were developed for easier operation and their larger capacity. In this study, the herpes simplex virus type-1 thymidine kinase (HSVtk) gene was cloned into the pHE700 amplicon vector to make an HE7tk vector and used for in vivo gene delivery. Human melanoma xenografts were established in athymic nude mice. Tumors were injected directly with HE7tk vector alone, HE7tk vector followed by ganciclovir (GCV), or a pHE700 amplicon vector carrying a green fluorescent protein (HE7GFP) gene followed by GCV. Efficient HSVtk transgene expression was found in the tumor 3 days after injection. Animals transduced with HE7tk followed by GCV had minimal tumor growth (P < .01 ). Animals that received either HE7tk vector without GCV or HE7GFP vector with GCV had some reduction in tumor growth compared to animals that were injected with buffer only. These data indicate that replication-defective HSV-1 amplicon vectors can be used effectively to deliver transgenes into solid tumors in vivo. PMID- 11916239 TI - Enhanced antitumor effect of EGF R-targeted p21WAF-1 and GM-CSF gene transfer in the established murine hepatoma by peritumoral injection. AB - One of the major obstacles in current cancer gene therapy is the lack of a gene delivery system with high efficiency and targetability. In this paper, a nonviral gene delivery system GE7, which was designed to target EGF receptor (EGF R) overexpressed on the surface of cancer cel Is through an EGF R-binding oligopeptide (GE7), was used for in vivo gene therapy in a murine subcutaneous hepatoma model. It was demonstrated that the GE7 system could target the reporter gene beta-gal to EGF R-expressing hepatoma cells with high efficiency after in vitro transfection and in vivo peritumoral injection. To improve the therapeutic effect elicited by single gene transfer, human cyclin-dependent kinase inhibitor gene p21WAF-1 and murine cytokine gene GM-CSFwere used simultaneously in peritumoral injection of the GE7/DNA polyplex. The results showed that combined gene transfer of p21WAF-1 and GM-CSF could inhibit the growth of pre-established tumor more effectively and prolong the survival time of hepatoma-bearing mice more significantly than the transfer of a single gene. Apoptosis in the tumor tissues were found when injected with the p21WAF1-DNA polyplex. Prominent inflammatory infiltration was observed in the tumor tissue transfected with the GM-CSF DNA polyplex. Our data demonstrate that the GE7 system-mediated, EGF R targeted cotransfer of p21WAF-1 and GM-CSF genes exhibit more potent antitumor effect by inducing tumor cell apoptosis and inflammatory responses. PMID- 11916240 TI - Secretion of interleukin-10 from murine colon carcinoma cells suppresses systemic antitumor immunity and impairs protective immunity induced against the tumors. AB - Interleukin-10 (IL-10) is a T helper type 2 (Th2) cytokine that suppresses Th1 mediated, cell-mediated immune responses and reciprocally enhances antibody mediated responses. Previous studies, however, demonstrated that forced expression of the IL-10 gene in tumor cells could unexpectedly produce antitumor effects. We then examined whether tumor-derived IL-10 could modulate systemic immune responses. Murine colon carcinoma (Colon 26) cells that were retrovirally transduced with the murine IL-10 gene (Colon 26/IL-10) were inoculated in syngeneic immunocompetent or T cell-defective nude mice. Growth of Colon 26/IL-10 tumors was augmented in immunocompetent and, to less extent, in nude mice compared with that of wild-type tumors developed in respective mice. Growth of wild-type tumors was accelerated to the same level as that of Colon 26/IL-10 tumors when wild type and Colon 26/IL-10 cells were respectively inoculated in different flanks of the same immunocompetent mice. This enhanced growth of wild type tumors was not observed in nude mice. Immunocompetent mice that had rejected IL-2- or IL-12-secreting Colon 26 cells developed protective immunity and became completely resistant to wild-type Colon 26 cells subsequently challenged. However, some of the mice that had rejected IL-2 or IL-12 producers developed Colon 26/IL-10 tumors inoculated thereafter. The present study showed that production of IL-10 from tumor cells impaired T cell- and non-T cell-mediated systemic antitumor immunity in hosts. PMID- 11916241 TI - Suppression of murine mammary carcinoma growth and metastasis by HSVtk/GCV gene therapy using in vivo electroporation. AB - The effectiveness of electroporation as a means of gene transfection, both in vitro and in vivo, was tested using the herpes simplex virus 1 thymidine kinase (HSVtk) gene in combination with ganciclovir (GCV) administration as therapy against murine mammary cancer. Approximately 80% of BJMC3879 metastatic mammary carcinoma cells, derived from MMTV-infected BALB/c mice, died as a result of HSVtk/GCV treatment 72 hours after the transfection; decreased DNA synthesis was also seen. Mammary tumors induced by inoculation of syngeneic mice with BJMC3879 cells were subsequently treated by direct injection of vector containing HSVtk (pHSVtk) alone, empty vector or saline alone twice a week. After each injection, the tumors were subjected to in vivo electroporation. Mice treated with pHSVtk or saline were intraperitoneally injected with GCV at 40 mg/kg five times a week. Significantly reduced tumor volumes were observed for the pHSVtk+GCV group in experimental week 2 and thereafter throughout the 2-month study. DNA synthesis was significantly decreased as well in the pHSVtk+GCV group compared with all other groups. Furthermore, metastasis to lymph nodes and lungs was significantly suppressed by HSVtk/GCV treatment. Expression of HSVtk in the tumors was confirmed by RT-PCR. Macrophage accumulations were frequently observed in the peripheries of necrotic regions in HSVtk/GCV-treated tumors, where levels of apoptosis were significantly higher than those observed in other groups. We therefore conclude that in vivo electroporation can result in efficient gene transfer and that the HSVtk/GCV prodrug system strongly suppresses tumor growth and metastases in this model. PMID- 11916242 TI - Aerosol delivery of PEI-p53 complexes inhibits B16-F10 lung metastases through regulation of angiogenesis. AB - Inhibition of pulmonary metastases poses a difficult clinical challenge for current therapeutic regimens. We have developed an aerosol system utilizing a cationic polymer, polyethyleneimine (PEI), for topical gene delivery to the lungs as a novel approach for treatment of lung cancer. Using a B16-F10 murine melanoma model in C57BL/6 mice, we previously demonstrated that aerosol delivery of PEI p53 DNA resulted in highly significant reductions in the tumor burden (P < .001) in treated animals, and also lead to about 50% increase in the mean length of survival of the mice-bearing B16-F10 lung tumors. The mechanisms of this antitumor effect of p53 are investigated in this report. Here, we demonstrate that the p53 transfection leads to an up-regulation of the antiangiogenic factor thrombospondin-1 (TSP-1) in the lung tissue and the serum of the mice. Furthermore, there is a down-regulation of vascular endothelial growth factor (VEGF) in the lung tissue and serum of the B16-F10 tumor-bearing mice treated with PEI-p53 DNA complexes, compared with untreated tumor-bearing animals. In addition, staining for von Willebrand factor (vWF), a marker for the angiogenic blood vessels, revealed that p53 treatment leads to a decrease in the angiogenic phenotype of the B16-F10 tumors. Immunohistochemistry for transgene expression reveals that the PEI-p53 aerosol complexes transfect mainly the epithelial cells lining the airways, with diffuse transfection in the alveolar lining cells, as well as, the tumor foci in the lung tissue. There was also some evidence of apoptosis in the lung tumor foci of animals treated with p53. The data suggest that aerosol delivery of PEI-p53 complexes leads to inhibition of B16-F10 lung metastases, in part by suppression of angiogenesis. PMID- 11916243 TI - A role for MAP kinase in the antitumor activity of a nucleoside analog. AB - Nucleoside analogs (NAs) have been used extensively in both antitumor and antiviral therapies. Their general mechanism of action has been postulated to result from incorporation into DNA, leading to disruption of DNA synthesis and DNA polymerase inhibition. To further explore the antitumor mechanisms of NAs we have evaluated ganciclovir (GCV), an NA antiviral agent, in herpes simplex virus thymidine kinase (HSV-TK) gene-modified tumor cells. This system allows specific evaluation of the antitumor effects of NAs because the antitumor effect is directly related to the phosphorylation of the prodrug GCV by the HSV-TK enzyme in the gene-modified tumor cells. We demonstrated that GCV incorporates into DNA and inhibits DNA polymerase, as has been observed in HSV-infected cells and with other antitumor NAs in tumor cells. A novel observation is that GCV activates MAP kinase within 1 hour of GCV exposure. This activation directly correlates with cytotoxicity, because inhibition of the MAP kinase extracellular regulated kinase (Erk) by PD98059, reversed GCV-mediated cytotoxicity. This effect appears to be specific to the Erk pathway, because inhibition of the p38 kinase with SB203580 had no effect on cytotoxicity. Further, GCV does not act as a DNA-damaging agent or activate general DNA-repair mechanisms, but does produce a number of metabolic disruptions, including a reversible decrease in NAD levels. These effects appear to be downstream of the earlier activation of Erk in this system, which may be a novel mechanism of action for GCV cytotoxicity in HSV-TK gene-modified tumor cells, and thus, needs to be further evaluated as the mechanism of tumor cell killing by other antitumor NAs. PMID- 11916245 TI - Adenoviral delivery of A-FOS, an AP-1 dominant negative, selectively inhibits drug resistance in two human cancer cell lines. AB - Activator protein-1 (AP-1) transcription factor has been linked to chemotherapeutic resistance. To assess the clinical efficacy of AP-1 inhibition toward reversing drug resistance, we have developed an adenovirus expressing a dominant negative that inhibits AP-1 DNA binding, namedAdA-FOS. We examined the consequence of AdA-FOS infection on two paired human cancer cell lines, each pair consisting of a parental cell and the drug- resistant derivative. The first pair of cells is the parental human ovarian cancer cell line A2780 and the cisplatin resistant A2780/CP70 cell line. The second pair of cells is the parental epidermal carcinoma cell line KB8 and the multidrug-resistant (mdr) KB85 cell line. Because of an association of up-regulated AP-1 activity with their drug resistance, these cell lines were considered good targets of AdA-FOS therapy. Following infection of the drug-sensitive and drug-resistant cells, we observed a significant decrease in cell viability of KB85 and A2780/CP70 cells at drug doses normally not lethal to the cell. The parental cell lines, A2780 and KB8 cells, were not similarly affected by AdA-FOS. This decrease in viability was specific to AdA-FOS as an adenovirus control (Advector) did not reverse drug resistance. Although the efficiency of AdA- FOS in therapy would need to be further analyzed with other cisplatin-resistant and mdr cell lines, these results suggest that AP 1 is a therapeutic molecular target and that inhibition of AP-1 DNA binding may be of clinical value in treating chemotherapeutic resistance. PMID- 11916244 TI - Local tumor irradiation augments the antitumor effect of cytokine-producing autologous cancer cell vaccines in a murine glioma model. AB - The combined therapeutic effect of cytokine-producing cancer cell vaccines and local radiotherapy was studied in a mouse glioma 261 (GI261) brain tumor model. Brain tumor-bearing mice were treated with cytokine (IL -4, IL-6, IL-7, GM-CSF, TNF-alpha, LIF, LT) producing vaccines made by in vitro transduction of GI261 cells with the corresponding adenoviral vectors. Vaccines producing either IL-4 or GM-CSF cured 20-40% of mice. The antitumor effect strongly depended on the secreted cytokine level. Vaccination therapy induced specific activation of cytotoxic T lymphocytes measured by cell-mediated cytotoxicity assay. Brain tumors were heavily infiltrated by CD4+ lymphocytes after treatment with IL-4- or GM-CSF-secreting cells. GM-CSF vaccination induced moderate CD8+ infiltration, as well. Depleting either CD4+ or CD8+ lymphocyte subsets abolished the anticancer effect of GM-CSF-expressing cells. Strong synergism was observed by combining cytokine vaccination (GM-CSF, IL-4, IL-12) with local tumor irradiation: about 80 100% of the glioma-bearing mice was cured. The high efficiency of combined treatment was maintained even under suboptimal conditions when neither of the modalities cured any of the mice alone. This suggests that vaccination therapy might open a new potential in the clinical treatment of high-grade gliomas when applied as adjuvant to existing treatment modalities. PMID- 11916246 TI - Purging of chronic myelogenous leukemia cells by retrovirally expressed anti-bcr abl ribozymes with specific cellular compartmentalization. AB - In patients with chronic myelogenous leukemia (CML), abnormal expansion of myeloid cells is maintained by expression of the p210(bcr-abl) fusion protein. Thus, this protein and its mRNA represent primary targets to inhibit proliferation of these cells. Here we describe the properties of a ribozyme against the bcr-abl mRNA, expressed as a fusion transcript with the human U1 small nuclear RNA or the adenovirus VA1 RNA and delivered to the cells through retroviral vectors. These fusion ribozymes are specifically localized in the nucleus or in the cytoplasm, respectively. Transduction of 32D-LG7 myeloid cells, whose growth is IL-3 independent thanks to deregulated bcr-abl expression, imposed strong negative selective pressure on cell growth and induced restoration of an IL-3-dependent phenotype. Although expressed at a level similar to that of the U1-fusion ribozyme, the cytoplasmic VA1 ribozyme was a more powerful inhibitor of p210(bcr-abl) gene expression. In cells transduced with the vector expressing this ribozyme, the levels of the bcr-abl transcript were reduced up to 10(4)-fold, the p210(bcr-abl) protein became undetectable, and the cells underwent massive apoptosis when cultured in the absence of IL-3. Transduction of primary hematopoietic cells obtained from bone marrow of patients with CML resulted in remarkable reduction of bcr-abl mRNA levels, starting a few days after transduction. These results show the feasibility and efficacy of vector expressed anti-bcr-abl ribozymes for purging of CML cells. PMID- 11916247 TI - Purified herpes simplex virus thymidine kinase retroviral particles: III. Characterization of bystander killing mechanisms in transfected tumor cells. AB - An important consequence of the suicide gene therapeutic paradigm is the phenomenon of bystander cell killing, the death of adjacent tumor cells not transduced with the thymidine kinase (TK) gene from herpes simplex virus (HSV) after treatment with the antiviral drug, ganciclovir (GCV). Evidence from quantitative in vitro assays of glioma cell lines suggest that both murine and human gliomas are similar in expressing high sensitivity to the bystander effect. In five of six glial tumors examined, the presence of only 5% of HSV-TK expressing transduced cells in the culture resulted in >90% tumor cell death/stasis after addition of GCV. Several lines of evidence support gap junction intercellular communication (GJIC) as important in the bystander effect. In vitro metabolic assays, performed with GCV in the medium, indicated that more tumor burden was reduced when culture conditions supported cell-cell contact of parental and HSV-TK-transduced cells. Additionally, a double dye transfer assay showed that cell communication through the gap junction is greatest for glioma, less for melanoma, and much less for colorectal carcinoma cell lines. In vitro metabolic assays with mixtures of TK+/TK- homologous tumor cells confirmed that glioma cell lines were more susceptible to bystander killing than melanomas. Assays with chimeric tumor mixtures of TK+/TK - cells showed that the level of the bystander killing obtained was characteristic of the TK-bystander cells. The in vitro findings were confirmed in vivo with GCV-treated homologous and chimeric tumors composed of TK+/TK- cells. Day 21 mean tumor volumes (MTVs) indicated the growths obtained were characteristic of the bystander activity reflective of the nontransduced cell population. Furthermore, nontransduced, high-GJIC cells in a chimeric tumor mass appeared to effectively bridge between transduced tumor cells and poorly communicating nontransduced cells. Finally, the importance of a gap junction protein, such as connexin-43, in facilitating the bystander effect was demonstrated with the HT29 low-GJIC cell line. When the TK-nontransduced cell population expressed connexin-43, a better bystander kill was achieved compared to the parental counterpart. PMID- 11916248 TI - In situ adenoviral interleukin 12 gene transfer confers potent and long-lasting cytotoxic immunity in glioma. AB - Interleukin 12 (IL-12) isa cytokine that promotesan antitumor Th1-type pattern of differentiation in mature naiveT cells. Despite its therapeutic success in multiple animal models of cancer, the utility of systemically administered recombinant cytokine has been limited by its toxicity. This has encouraged the development of local IL-12 delivery systems through gene transfer. To determine the effect of local adenoviral delivery of IL- 12 on glioma immunogenicity, mice bearing GL-26 gliomas in the right corpus striatum were treated with direct intratumoral administration of AdmIL-12, AdLacZ, or normal saline. Survival was significantly prolonged in AdmIL-12-treated animals and immunohistochemistry demonstrated robust CD4+ and CD8+ T-cell infiltration in these mice compared to the two control groups. Glioma-infiltrating T lymphocytes from mice that received AdmIL-12 also demonstrated relatively increased, albeit statistically nonsignificant tumor killing. Based on IL-12's known ability to enhance Th1-type cytotoxic antitumor immune responses, we postulate our findings to be a result of localized induction of tumor immunity. PMID- 11916249 TI - Lipofection of early passages of cell cultures derived from murine adenocarcinomas: in vitro and ex vivo testing of the thymidine kinase/ganciclovir system. AB - Early passages of cultured cells derived from four spontaneous Balb/c murine adenocarcinomas were used to explore the feasibility of a nonviral HSVtk-based suicide gene therapy system. After lipofection with pCMVtk, the transiently HSVtk expressing P07 (lung), M3, M05, and M38 (mammary gland) cells were, respectively, about 130-, 30-, 120-, and 170-fold more sensitive to ganciclovir (GCV) in vitro than their respective controls. Eighty percent of Balb/c mice subcutaneously inoculated with ex vivo pCMVtk-lipofected P07 cells, followed by intraperitoneal GCV injection for 7 days, displayed a complete inhibition of tumor growth for over 70 days. Control animals started to display tumors 13 days after inoculation. We present evidence showing that early passages of cultured tumor cells can efficiently express lipofected genes and that they are sensitive to the lipoplex-mediated HSVtk/GCV system. PMID- 11916250 TI - Flow-through polymerase chain reactions in chip thermocyclers. AB - The miniaturization of analytical devices by micromachining technology is destined to have a major impact on medical and bioanalytical fields. To meet the current demands for rapid DNA amplification, various instruments and innovative technologies have been introduced by several groups in recent years. The development of the devices was extended in different directions and adapted to corresponding applications. In this review the development of a variety of devices and components for performing DNA amplification as well as the comparison of batch-process thermocyclers with reaction chambers and flow-through devices for different purposes are discussed. The main attention is turned to a flow device concept for thermocycling using microfabricated elements for local heat flow management, for which simulations and considerations for further improvement regarding design, material choice and applied technology were performed. The present review article mainly discusses and compares thermocycling devices for rapid thermocycling made of silicon or of silicon and glass with a short excursion to the possibility of plastic chip devices. In order to perform polymerase chain reactions (PCRs) in the microreactors, special attention must be paid to the conditions of the internal surfaces. For microchips, surface effects are generally pronounced because the surface to volume ratio increases upon miniaturization. Solutions for solving this problem are presented. We propose an overview of layouts for batch-process thermocyclers with different parallelization of reaction chambers and also of different designs of continuous flow thermocycling chips, paying particular attention to the parameters which influence the efficiency of such chip devices. Finally we point out some recent issues for applications in the field of clinical diagnostics. PMID- 11916251 TI - Sampling and monitoring in bioprocessing using microtechniques. AB - In this review we describe aspects of interactions between bioreactors and analytical systems including microsystems. Principles of bioprocess monitoring are summarized, before we focus on the miniaturization of sampling systems guaranteeing bioprocess sterility and providing analytical systems with a liquid sample. The application of negative dielectrophoresis as a new principle for cell retention in a sampling system is described followed by theoretical aspects and results. Properties of micromachined silicon membranes as filters for sampling systems and for biosensor protection are discussed. PMID- 11916252 TI - Nanotiterplates for combinatorial chemistry. AB - Miniaturization has grown to be a driving force in modern chemical and biochemical laboratories. Combinatorial explosion demands for new pathways for the synthesis and screening of new substances which can act as leads in drug discovery. Highly parallelized automata that can handle the smallest amounts of substances are needed. However, the development is not always straightforward since new problems also arise in miniaturization, e.g. increasing importance of surface properties of utilized devices and evaporation of liquids. This paper reports on recent developments on the field of miniaturized reaction vessels called nanotiterplates. A survey on fabrication technologies as well as applications of nanotiterplates is given. Special emphasis is given to results of the development of an automaton for miniaturized synthesis and screening. Besides the mere fabrication of nanotiterplates with integrated microsieve bottom membranes, examples of applications in chemical synthesis and bio-assays are given. Further topics are the characterization and specific adaption of surface properties and investigations on the evaporation of solvents and measures for prevention. PMID- 11916253 TI - Microfluidic components for protein characterization. AB - The use of microfluidic components to create an analytical toolbox for the very rapidly growing field of proteomics is described. This toolbox provides novel generic analytical solutions that are highly adaptable for analysis of various biomolecules, ranging from high to low abundant. The components are fabricated using silicon micromachining and consist of a microchip immobilised enzyme reactor (microIMER), a piezoelectric microdispenser and high-density nanovial target plates. This microtechnology based platform interfaces matrix-assisted laser desorption/ionisation time-of-flight mass spectrometry (MALDI TOF-MS) to a wide range of upstream sample handling and/or analytical techniques. Examples of applications such as rapid on-line digestion (12 s) and sample preparation of proteins, interfacing to capillary liquid chromatography (100 attomol sensitivity), and in-vial chemistry on femtomol amounts of sample are presented. PMID- 11916254 TI - What is a microreactor? It is a miniature version of the traditional, large-scale reactor that most people are familiar with. PMID- 11916255 TI - Polymer based micro-reactors. AB - In this paper, we describe the fabrication technologies necessary for the production of polymer-based micro-fluidic devices. These technologies include hot embossing as a micro-structuring method as well as so-called back-end processes to complete the micro-devices. Applications such as capillary electrophoresis, micro-mixers and nanowell plates are presented. PMID- 11916256 TI - Characterization of the region encompassing the human lysyl oxidase locus. AB - A 46,823 bp region of human chromosome 5q23.1 encompassing the seven-exon lysyl oxidase gene was characterized at the primary sequence level. Approximately 17.4% of this region is comprised of repetitive elements. The gene colocalizes with microsatellite marker D5S467. It is flanked by two candidate nuclear matrix association regions (MARs). The 5' MAR centered at position 12,500 is of the AT rich and curved DNA class. This is followed by a large CpG island containing fifty-seven putative regulatory elements which extend from just upstream of exon 1 to intron 2. The larger 3' MAR, spans position 35,050-39,750 and is characterized by a TG-rich kinked structure that also contains a topoisomerase II binding site. Based on these results model of the transcriptional regulation of the lysy/oxidase gene is presented. PMID- 11916257 TI - A 6940 bp DNA fragment from Desulfovibrio gigas contains genes coding for lipoproteins, universal stress response and transcriptional regulator protein homologues. AB - The nucleotide sequence of a 6940 bp DNA fragment from Desulfovibrio gigas, containing seven ORFs was determined. ORF-1 encodes a probable lipoprotein having high similarities with lytic transglycosylases. ORF-2 encodes a polypeptide that does not show homologies with proteins deposited in the database, but it contains the consensus pattern of class II aminoacyl-tRNA synthetases. The putative protein encoded by ORF-3 possesses high similarities with universal stress response proteins from the UspA family. Northern blot analysis of ORF-3 shows that it is constitutively and abundantly expressed. ORF-4 encodes a probable helix-turn-helix-containing DNA-binding protein, given the presence of the helix turn-helix motif, characteristic of this class of proteins. Its N-terminal region has high identity with its counterparts from proteins belonging to the RRF2 family. Northern blot analysis shows that ORF-4 is transcribed as a single mRNA in contrast to its orthologue from Desulfovibrio vulgaris. ORF-5 encodes a putative fusion protein as its N- and C-termini show significant homologies with molybdenum formylmethanofuran dehydrogenase and molybdopterin biosynthesis proteins, respectively. ORF-7 encodes a prokaryotic lipoprotein having homologies with multidrug efflux and DNA damage-inducible proteins, and it is constitutively and abundantly expressed. PMID- 11916258 TI - Comparison of the coding sequence of the platelet-activating factor receptor gene in three species. AB - The actions of platelet-activating factor (PAF, 1-O-alkyl-2-acetyl-sn-glycero-3 phosphocholine) are mediated through the PAF receptor (PAFr), which is a member of G-protein coupled superfamily of receptors. Our laboratory has data showing PAF has a role(s) in reproduction in domestic animals. Porcine, bovine and caprine PAFr genes cloned in BAC vectors were sequenced. Each PAFr coding sequence (cds) in these three species is 1029 nucleotides long and contains no intervening sequences. The deduced amino acid sequences (AAS) appear to contain seven putative transmembrane domains with an extracellular N-terminus in each species. There is a common glycosylation site at the fourth asparagine residue of N-terminus. In the tail of each deduced amino acid sequence, five to six serines and five threonine residues could act as phosphorylation sites, which play an important role in rapid receptor desensitization. The degree of homology of the three species is from 89 to 96% in nucleotide sequences (NtS), and 87-96% in identities (I) and 94-97% in positives (P) in amino acid sequences (AAS). The degree of homology with human, guinea pig, mouse and rat is 84-87, 82-88 and 83 88% in NtS, 77-84 (I) or 85-90 (P), 77-84 (I) or 85-90 (P) and 75-83 (I) or 87 90% (P) in AAS for caprine, bovine and pig, respectively. Southern blotting results suggested that the PAFr gene exists as a single copy in the genome of pig. PMID- 11916259 TI - cDNA cloning of proglucagon from the stomach and pancreas of the dog. AB - In human and rat, tissue-specific proteolytic processing of identical proglucagon precursors yield tissue-specific proglucagon-derived peptides. In contrast, in many non-mammalian vertebrates alternative mRNA splicing yields different proglucagon precursors in different tissues. Thus alternative mRNA splicing, in part, limits the choices of proglucagon-derived peptides that can be produced by proteolytic processing. Stomach proglucagon mRNAs from the rainbow trout and Xenopus laevis were found not to encode the proglucagon-derived peptide glucagon like peptide 2 (GLP-2). To determine if the absence of GLP-2 was a general feature of stomach proglucagons we isolated and characterized proglucagon cDNAs from the stomach and the pancreas of the dog, a mammal that expresses the proglucagon gene in the stomach. A major proglucagon transcript of about 1100 bases and a minor transcript of about 800 bases were identified in both stomach and pancreas. The coding sequences of both the stomach and pancreatic proglucagon transcripts were identical. Therefore, tissue-specific proteolytic processing, and not alternative mRNA splicing, must regulate the production of tissue specific proglucagon-derived peptides from the stomach of the dog. PMID- 11916260 TI - Molecular analysis of the 5' region of human ribosomal transcription factor UBF. AB - Upstream binding factor, UBF, is a nucleolar autoantigen involved in the transcription of ribosomal DNA genes. Previously, human genomic clones served to demonstrate that an alternative pre-mRNA splicing of a single gene is used to form UBF1 and UBF2. Here, to complete characterizing the 5'end genomic organization of this nucleolar transcription factor, lambda clones containing the human UBF gene were isolated from a human placenta genomic library using a hamster UBF cDNA as a probe. An additional PCR product was isolated from HeLa genomic DNA to cover the first translated 60 nt of the gene containing the ATG initiation codon. We have also determined the transcription start site of the gene by primer extension analysis at nt 188 upstream from the start ATG codon. It served first, to identify an untranslated initial exon on the UBF gene covering the first 121 nt of human UBF cDNA, and also to establish the sequence of the proximal promoter. The human UBF promoter lacks a TATA and CAAT boxes but contains multiple binding sites for SP1, AP1, AP2, TFIID, NF-1 and a single site for NFAT-1. Consequently we have defined the first five exons of the human UBF gene covering 7.5kb. The complete gene now consists of 20 exons with intervening sequences and spans approximately 15kb of DNA. PMID- 11916261 TI - Relationship of transcription of Drosophila melanogaster gene CG11327 and the gene for a thrombospondin homologue (DTSP). AB - Because of the close proximity of D. melanogaster gene CG11327 and the gene for a thrombospondin homologue (DTSP), the relationship between transcription of the two genes has been investigated. As part of this study, the cDNA sequence of gene CG11327 was determined, and structural features of the encoded protein were characterized. The CG11327 open reading frame specifies a polypeptide of 475 amino acid residues, molecular weight 53,666. The highly acidic protein has an aspartic acid + glutamic acid content of 23.2%. Weak similarities to other proteins suggest that it may function as a transcription factor or structural protein of the cell cycle. The existence of overlapping transcripts was found to be a significant feature of transcription of gene CG11327 and the DTSP gene. The 3' end of the DTSP gene overlaps the 5' end of gene CG11327 by at least 177bp, and the region of overlap includes exon 13 and the 3' UTR of the DTSP gene. PMID- 11916262 TI - Homology study of two polyhydroxyalkanoate (PHA) synthases from Pseudomonas aureofaciens. AB - Recently, we have cloned and analyzed two polyhydroxyalkanoate (PHA) synthase genes (phaC1 and phaC2 in the pha cluster) from Pseudomonas aureofaciens. In this report, the deduced amino acid (AA) sequences of PHA synthase 1 and PHA synthase 2 from P. aureofaciens are compared with those from three other bacterial strains (Pseudomonas sp. 61-3, P. oleovorans and P. aeruginosa) containing the homologous pha cluster. The level of homology of either PHA synthase 1 or PHA synthase 2 was high with each enzyme from these three bacterial strains. Furthermore, multialignment of PHA synthase AA sequences implied that both enzymes of PHA synthase 1 and PHA synthase 2 were highly conserved in the four strains including P. aureofaciens. PMID- 11916263 TI - Molecular cloning and sequence analysis of C57BL/6 mouse contrapsin cDNA. AB - Contrapsin is a member of the serpin superfamily and inhibits trypsin much more strongly than alpha1-antiproteinase. Mouse and rat contrapsins, however, have similarity in sequence to human alpha1- antichymotrypsin. In order to test the hypothesis that reactive site regions of contrapsin family evolved under strong selective pressure, cDNA sequence of C57BL/6 mouse contrapsin was determined and compared with that of ICR mouse. The cDNA sequence of C57BL/6 mouse contrapsin was found to contain an open reading frame encoding polypeptide consisting of 418 amino acid residues. The work reported in this paper shows that the reactive site is not hypervariable as compared with the rest of molecule. PMID- 11916264 TI - Characterization of a processed pseudogene of human psiHSP40 on chromosome 2q32. AB - A pseudogene for the human Hsp40 gene has been characterized (psiHSP40). The pseudogene sequence shows 90% similarity to the human Hsp40 mRNA at the nucleotide level. No introns were found in the region corresponding to the human Hsp40 cDNA, and two direct repeats flank this same region. Because of these features, the pseudogene can be classified as a processed pseudogene. PsiHSP40 was assigned to chromosome 2q32 by in situ hybridization. This is the first report of a pseudogene for a member of the DnaJ (Hsp40) family protein gene. PMID- 11916265 TI - Relationship between the sialic acid content of low-density lipoprotein (LDL) and autoantibodies to oxidized LDL in the plasma of healthy subjects and patients with atherosclerosis. AB - To determine whether the sialic acid (SA) content of the low-density lipoprotein (LDL) is related to the plasma concentration of autoantibodies to oxidized LDL (oxLDL), we measured the SA content of LDL and the concentrations of oxLDL and autoantibodies to oxLDL in plasma of 20 apparently healthy subjects and 20 patients with advanced coronary atherosclerosis. In the healthy subjects the SA content of LDL correlated positively with plasma concentration of autoantibodies to oxLDL. In agreement with the literature the decreased SA content of LDL was associated with an increased fraction of oxLDL; a decreased fraction of oxLDL was associated with an increased plasma concentration of autoantibodies to oxLDL. In the patients the SA content of LDL and plasma concentrations of oxLDL and autoantibodies to oxLDL were not related. We conclude that the SA content of LDL correlates positively with plasma concentration of autoantibodies to oxLDL in healthy subjects. However, this association may vary depending on the stage of atherogenesis. Although our results suggest dependence of LDL SA content on the clearance of oxidatively modified (desialylated and oxidized) LDL from blood by autoantibodies to oxLDL, the mechanisms regulating the SA content of LDL await further studies. PMID- 11916266 TI - Growing significance of myeloperoxidase in non-infectious diseases. AB - Myeloperoxidase (MPO) is a glycoprotein released by activated polymorphonuclear neutrophils, which takes part in the defense of the organism through production of hypochlorous acid (HOCl), a potent oxidant. Since the discovery of MPO deficiency, initially regarded as rare and restricted to patients suffering from severe infections, MPO has attracted clinical attention. The development of new technologies allowing screening for this defect has permitted new advances in the comprehension of underlying mechanisms. Apart from its implications for host defense, the expression of MPO restricted to myeloid precursors makes MPO mRNA a good marker of acute myeloid leukemia. In addition, during the last few years, involvement of MPO has been described in numerous diseases such as atherosclerosis, lung cancer, Alzheimer's disease and multiple sclerosis. Both strong oxidative activity and MPO genetic polymorphism have been involved. This review summarizes the broad range of diseases involving MPO and points out the possible use of this protein as a new clinical marker and a future therapeutic target. PMID- 11916267 TI - The presence of heparin-platelet factor 4 antibodies as a marker of hypercoagulability during hemodialysis. AB - We have assessed the prevalence of heparin-platelet factor 4 antibodies in a group of 100 patients on chronic intermittent hemodialysis with repeated exposure to heparin in order to establish the clinical significance of an immunological response to heparin. Heparin-induced platelet reactivity was studied in a subgroup of 86 patients. Routine laboratory parameters such as plasma C-reactive protein, albumin and immunoglobulins were included. Clinical endpoints were thrombosis in blood access, change of blood access, other thromboembolic events, bleeding complications, falling platelet count, and ischemic vascular disease or death within 1 year of blood sampling. Blood access thrombosis was a frequent complication (26%), often leading to change of dialysis blood access (21%). Antibodies were detected in 3-6% of hemodialysis patients and antibody levels correlated to blood access thrombosis and change of access (p<0.05). An unexpected finding was that of increased heparin-induced platelet reactivity in 36% of the control group of 24 uremic patients prior to dialysis or exposure to heparin. The findings suggest that antibodies to heparin-platelet factor 4 may contribute to hypercoagulability during hemodialysis, leading to thrombotic complications in affected patients. PMID- 11916268 TI - The n-3 and n-6 polyunsaturated fatty acid composition of plasma phospholipids in pregnant women and their infants. relationship with maternal linoleic acid intake. AB - The availability of long-chain polyunsaturated fatty acids during infancy has been related to neonatal growth and development. Fatty acid concentration at birth is an important predictor of postnatal level. The primary aim of this study was to provide a description of the distribution of n-3 and n-6 polyunsaturated fatty acids in the plasma phospholipid fraction of pregnant women remaining on a Western-style diet and their neonates. The plasma phospholipid polyunsaturated fatty acid composition was determined by gas-liquid chromatography in 889 mother infant pairs. Blood samples were taken during the first, second and third trimester of pregnancy, at delivery, and from the umbilical vein at birth. Mean (+/- SD) fatty acid concentrations are reported in mg/l and as percentage of total fatty acids (% wt/wt). In addition, the 10th, 25th, 50th, 75th and 90th percentiles are given. The distribution of docosahexaenoic acid (22:6n-3) and arachidonic acid (20:4n-6) concentrations in umbilical plasma phospholipids is also reported as a function of gestational age and maternal linoleic acid intake during pregnancy. This data can be be used as a reference for future studies and may aid in identifying term infants with a relatively low long-chain polyene status at birth. PMID- 11916269 TI - Quantification of sirolimus by liquid chromatography-tandem mass spectrometry using on-line solid-phase extraction. AB - Quantification of the new immunosuppressant sirolimus (syn. rapamycin; Rapamune) in whole blood by chromatography is essential for its clinical use since no immunoassay is available although monitoring is mandatory. Here we report on a rapid and convenient liquid chromatography (LC-tandem mass spectrometry method and describe our practical experience with its routine use. Whole blood samples were hemolyzed and deproteinized using an equal volume (150 microl) of a mixture of methanol/zinc sulfate solution containing the internal standard desmethoxy rapamycin. After centrifugation, the clear supernatants were submitted to an on line solid-phase extraction procedure using the polymeric Waters Oasis HLB material, with elution of the extracts onto the analytical column in the back flush mode by column switching. For analytical chromatography a RP-C18 column was used with 90/10 methanol/2 mM ammonium acetate as the mobile phase. A 1:10 split was used for the transfer to the mass spectrometer, a Micromass Quattro LC-tandem mass spectrometry system equipped with a Z-spray source and used in the positive electrospray ionization mode. The following transitions were recorded: sirolimus, 931>864 m/z, and desmethoxy-rapamycin (I.S.), 901>834 m/z. The analytical running time was 5 min, including on-line extraction. The method has a linear calibration curve (r>0.99; range 1.6-50 microg/l) and is rugged and precise with monthly CVs <7% at a sirolimus concentration of 13.1 microg/l in routine use; the instrumentation proved to be reliable and required minimal maintenance. Clin Chem PMID- 11916270 TI - The effects of affinity-purified anti-DNA antibodies from patients with systemic lupus erythematosus on the fluorescent antinuclear antibody assay using HEp-2 cells. AB - The aim of this study was to clarify the effects of anti-dsDNA antibodies on the titer and the nuclear staining pattern(s) in a fluorescent antinuclear antibody (FANA) assay using HEp-2 cells. Anti-dsDNA derived from 14 patients with systemic lupus erythematosus (SLE) was individually affinity-purified. The anti-dsDNA titer of the purified anti-dsDNA solution was measured by radioimmunoassay (RIA) or by enzyme-linked immunosorbent assay (ELISA). In the FANA assay, the anti dsDNA solution was diluted in a stepwise manner and its titer was expressed by the endpoint dilution. The nuclear staining pattern in the anti-dsDNA solution was examined at the 1:5 and 1:20 dilutions and at the endpoint dilution. The anti dsDNA titers of the affinity-purified anti-dsDNA solution were high enough (13 to 126 IU/ml) to be measured by RIA. However, the antinuclear antibody (ANA) titers of this solution were relatively low: 1:20 to 1:320. In the study of nuclear staining the peripheral pattern was observed in nine of the 14 cases at a 1:5 dilution. However, at the endpoint dilution, all cases exhibited the homogeneous pattern. These findings indicate that in the FANA assay using HEp-2 cells, 1) although serum samples show high anti-dsDNA titers by RIA or by ELISA, the antibodies' direct contribution to ANA titers is limited, and 2) when samples reveal a homogeneous staining pattern at the endpoint dilution, this suggests the presence of anti-dsDNA. PMID- 11916271 TI - Gender differences in C-reactive protein concentrations-confirmation with two sensitive methods. AB - Two new automated, highly sensitive methods for the measurement of C-reactive protein (CRP) were compared using blood samples from active and retired health care workers (116 males, 114 females). The regression equation was y=1.115x 0.0267 forthe Kamiya K assay immunoturbidimetric method (x) vs. the Behring BNII nephelometric method (y) with medians (IQR) of 0.156 (0.081 to 0.276) and 0.150 (0.060 to 0.240) mg/dl, respectively. Significant differences in hsCRP values between males and females were observed (p=0.013 and p=0.0018 for Behring and Kamiya methods, respectively). This gender difference in CRP levels was confirmed by analysis of serum samples from 1,114 individuals (639 males and 475 females) obtained through Mayo Medical Laboratories reference services, which represents participants from across the United States. The gender difference, which was confirmed (p<0.0001), was independent of age differences as assessed by multiple regression analysis. The observed gender differences have important implications for the establishment of cut-off points for cardiovascular risk stratification. PMID- 11916272 TI - Serum cholinesterase activity in patients with burns. AB - Serum cholinesterase activities were measured in 300 patients with thermal injuries. The samples were drawn immediately upon admission and thereafter daily until the time of the patients' discharge or death. According to the burn severity a characteristic decrease was noted during the first days. The decline during the first 24 hours as well as its value (measured 24 hours after admission) was found to be correlated with the total body surface area burned. In all patients the decrease in cholinesterase activity persisted for days, but in patients with inhalation trauma this decrease was significantly greater than in patients without lung injury. It seems that serum cholinesterase activity reflects not only the presence of an inhalation trauma. The cholinesterase measurement seems to be also a good method to observe the course of illness after a burn injury. Our data suggest a correlation between cholinesterase activity and morbidity. The inhalation trauma aggravates the thermal trauma. PMID- 11916273 TI - Serum lipids and apolipoproteins in patients with psoriasis. AB - Psoriasis is characterized by defects in the normal cycle of epidermal development that lead to epidermal hyperproliferation, altered maturation of skin cells, vascular changes and inflammation. Also, psoriasis has been associated with an abnormal plasma lipid metabolism. Changes in plasma lipid and lipoprotein composition in patients with psoriasis may be the reason for the increased risk of atherosclerosis in these patients. We determined serum concentrations of lipids, lipoproteins and apolipoprotein Al and B (apo A1 and apo B) in 72 patients with psoriasis and 30 age matched controls. Serum lipoprotein (a) (Lp(a)), apo A1 and apo B were measured by immunoprecipitation assays, and the lipids and other biochemical parameters by enzymatic methods. Serum Lp(a) and triglyceride (TG) were significantly higher in patients with psoriasis than in healthy control subjects (p<0.01 for both). Apo B was also found to be higher in the patient group, but the difference was not significant. The levels of total cholesterol (TC), high-density lipoprotein cholesterol (HDL-C), low-density lipoprotein cholesterol (LDL-C) and apo A1 did not differ significantly from those of the controls. These observations imply that serum Lp(a) and TG concentrations may play a role as risk factors for atherosclerotic disease in patients with psoriasis. PMID- 11916274 TI - Reference intervals for a complete blood count determined on different automated haematology analysers: Abx Pentra 120 Retic, Coulter Gen-S, Sysmex SE 9500, Abbott Cell Dyn 4000 and Bayer Advia 120. AB - We processed 317 samples from healthy adult volunteers for a complete blood count, including leukocyte differentials and reticulocyte parameters, through five new-generation haematology analysers: Abx Pentra 120 Retic, Coulter Gen-S, Sysmex SE 9500, Abbott Cell Dyn 4000 and Bayer Advia 120. From these data non parametric 2.5-97.5 percentile reference intervals were calculated for all parameters on all analysers. Some differences were found compared with previously reported reference intervals. Reference intervals for platelet parameters and reticulocytes agreed with these usually accepted. For red blood cell parameters, including haemoglobin and haematocrit, and white blood cell count, including absolute white blood cell differentials, our calculated reference intervals were in agreement with less frequently cited earlier reports, but were lower compared to the usually accepted reference intervals. PMID- 11916275 TI - Science, alchemy and light: paintings by Joseph Wright of Derby. AB - This article considers two paintings by the English painter, Joseph Wright of Derby (1734-97), 'An Experiment on a Bird in the Air Pump' and 'The Alchemist in Search of the Philosopher's Stone' as painterly reflections on the early industrial culture. Particularly interesting in the "Experiment on a Bird" is the broad spectrum of spectator's reactions to the experiment; this can be related to the contemporary debate on the contextualization of science. PMID- 11916276 TI - Approved IFCC reference method for the measurement of HbA1c in human blood. AB - HbA1C is the stable glucose adduct to the N-terminal group of the beta-chain of HbA0. The measurement of HbA1c in human blood is most important for the long-term control of the glycaemic state in diabetic patients. Because there was no internationally agreed reference method the IFCC Working Group on HbA1c Standardization developed a reference method which is here described. In a first step haemoglobin is cleaved into peptides by the enzyme endoproteinase Glu-C, and in a second step the glycated and non-glycated N-terminal hexapeptides of the beta-chain obtained are separated and quantified by HPLC and electrospray ionisation mass spectrometry or in a two-dimensional approach using HPLC and capillary electrophoresis with UV-detection. Both principles give identical results. HbA1c is measured as ratio between the glycated and non-glycated hexapeptides. Calibrators consisting of mixtures of highly purified HbA1c and HbA0 are used. The analytical performance of the reference method has been evaluated by an international network of reference laboratories comprising laboratories from Europe, Japan and the USA. The intercomparison studies of the network showed excellent results with intra-laboratory CVs of 0.5 to 2% and inter laboratory CVs of 1.4 to 2.3%. Possible interferences have been carefully investigated. Due to the higher specificity of the reference method the results are lower than those generated with most of the present commercial methods which currently are calibrated with unspecific designated comparison methods. The new reference method has been approved by the member societies of the International Federation of Clinical Chemistry and Laboratory Medicine and will be the basis for the future uniform standardization of HbA1c routine assays worldwide. PMID- 11916277 TI - Serum paraoxonase activity and the extent of lipid peroxidation are not affected by increased levels of human apolipoprotein A-I: studies in transgenic mice. AB - The present study analyzed the effect of increased concentrations of human apolipoprotein (apo) A-I in transgenic mice serum on paraoxonase activity and on lipid peroxidation. In the transgenic mice serum, in comparison to control (non transgenic) C57BL/6 mice, we found high concentrations of human apoA-I and high density lipoprotein (HDL)-cholesterol, but serum lipid peroxidation (basal and free radical-induced) and serum paraoxonase activity were similar in the two mouse groups. Comparing the individual results, no significant correlation was found between free radical-induced serum lipid peroxidation and apoA-I concentrations. Serum paraoxonase activity also did not correlate with serum concentrations of human apoA-I. However, a significant inverse relationship (R2=0.75) was observed between the individual values of paraoxonase activity and free radical-induced lipid peroxidation in both mouse groups. Direct analysis of the effect of pure human apoA-I and paraoxonase (using the specific paraoxonase inhibitor PD-92770) on lipid peroxidation also revealed that paraoxonase, but not apoA-I, protects serum lipids from oxidation. We thus conclude that the increased human apoA-I concentration in the mouse serum neither affect serum paraoxonase activity, nor protects against lipid peroxidation, whereas paraoxonase significantly inhibits serum lipid peroxidation. PMID- 11916278 TI - Characterization of sex hormone-binding globulin isoforms in hypothyroid women. AB - Liver sex hormone-binding globulin (SHBG) biosynthesis is regulated by triiodothyronine (T3). This regulation is responsible for increased serum SHBG concentrations in hyperthyroid states. However, in hypothyroidism, normal SHBG levels are frequently found. To understand this we have characterized circulating SHBG isoforms according to their sialic acid content, which determines its half life, in euthyroid and hypothyroid women. Six euthyroid (aged 56 +/- 8 years) and five hypothyroid women (51 +/- 13 years) were studied. Their body mass index (BMI) range was 20-25. Hypothyroidism diagnosis was based on clinical findings, elevated basal thyrotropin (TSH) and decreased T3 and thyroxine (T4) values. Total SHBG was measured by radioimmunoassay (RIA) and SHBG isoforms were isolated using preparative isoelectrofocusing. For comparisons, two-tailed t test was applied. No statistical difference was found between the total SHBG levels of hypothyroid and euthyroid postmenopausal women. Three groups of SHBG isoforms were isolated in the euthyroid group: S(I): pl: 5.0-5.2: 20% +/- 4%, S(II) : pl 5.2-5.4: 50% +/- 3% and S(III): pl 5.4-5.6: 29% +/- 4%. In hypothyroid patients, although the three groups of isoforms were isolated in the same pH range, S(I) and S(II) proportions were different (p < 0.001) when compared to normal women: S(I): 34% +/- 4%, S(II): 33% +/- 9.9% and S(III): 29% +/- 5.7%. These results show that hypothyroid patients have a higher proportion of more acidic SHBG isoforms. This variation may explain the normal levels of serum SHBG observed in hypothyroidism. PMID- 11916279 TI - A GC box in the human sodium iodide symporter gene promoter is essential for full activity. AB - We previously reported that the human sodium iodide symporter (hNIS) 5'-flanking region between -603 and -415 is essential for full expression. In this study, we further localized sequences within this region required for the basal expression of the hNIS promoter and identified a functional GC box. Activity of the hNIS promoter was assessed by transient transfection of luciferase reporter gene constructs with progressive 5' deletions into the human papillary thyroid cancer cell line BHP 2-7. Deletion from -603 to -535 enhanced promoter activity, further deletion to -469 decreased promoter activity, and deletion to -415 nearly abolished promoter activity. The DNA sequence within this critical 55 bp region from -469 to -415 contains a GC box. Introduction of mutations into the GC box of the deletion constructs -603, -535 and -469 decreased promoter activity in both thyroid cells (BHP 2-7 and rat thyroid cell FRTL-5) and nonthyroid cells (human prostate cancer cell LNCaP-2 and human breast cancer cell MCF-7). The magnitude of reduction for the -603 mutation construct was significantly greater than that for the -469 mutation construct in thyroid cells compared to non-thyroid cells. In vitro transcription using nuclear extracts isolated from HeLa cells was reduced from DNA templates with the GC box mutation and nearly abolished from templates with 5' deletion to -415. Identification of proteins interacting with the GC box was performed by gel retardation assays with or without Sp1 specific antibodies. Sp1 and an "Sp1-like" protein bound to the wild-type GC box sequence but not to the GC box mutant. In summary, the GC box is a positive regulatory element in the hNIS basal promoter. PMID- 11916280 TI - Evidence for a simplified view of autoantibody interactions with the thyrotropin receptor. AB - Recently, many exceptions have been reported that undermine the concept that the epitopes for thyroid-stimulating autoantibodies (TSAb) and thyrotropin-blocking autoantibodies (TBAb) lie within the N-terminal and C-terminal portions of the thyrotropin receptor (TSHR) ectodomain, respectively. Here we have used a new approach to examine the issue by attempting to neutralize autoantibodies with the purified, N-terminal 289 amino acids of the TSHR ectodomain (TSHR-289), essentially the A subunit. Immunoglobulin G (IgG) with TSAb activity from 10 patients with Graves' disease was preincubated with or without purified active or inactive TSHR-289. Active, but not inactive, TSHR-289 completely neutralized TSAb activity in all sera. We then tested the ability of active TSHR-289 to neutralize TBAb activity in two rare hypothyroid patients with pure TBAb activity lacking agonist activity. IgG from both patients was extremely potent in the TBAb assay. Unlike with TSAb, preincubation of the IgG with a large excess of active TSHR-289 (20 microg/mL) revealed a remarkable divergence. TBAb activity from one patient was totally neutralized whereas in the other patient TBAb activity was totally unaffected. In conclusion, using a new approach, we confirm that the major portion of the TSAb epitope in the 418 amino acid ectodomain lies upstream of residue 289 (within the A subunit). In contrast, TBAb that cause hyperthyroidism can be more heterogeneous, with epitopes that lie largely upstream (A subunit) or downstream (B subunit) of residue 289. PMID- 11916281 TI - Dosimetrically determined doses of radioiodine for the treatment of metastatic thyroid carcinoma. AB - In the absence of definitive studies relating radioiodine dose to outcomes, selection of a dose of radioiodine to treat metastatic thyroid carcinoma is problematic, and several approaches have been used. These include empiric fixed doses and doses used on dosimetric approaches specific for each patient. This paper is a review of the rationale and technique for dosimetrically-determined doses of radioiodine for the treatment of metastatic thyroid carcinoma. This review (1) discusses the alternatives for selection of a dose, (2) discusses the two major approaches for determining radioiodine doses dosimetrically, (3) briefly reviews several modifications of these approaches, (4) reviews the literature regarding the results, (5) discusses the side effects of these different approaches, and (6) concludes with recommendations for patient management and future research. This review does not address use of dosimetrically-determined doses of radioiodine for the initial ablation of thyroid tissue postoperatively. PMID- 11916282 TI - The effect of methimazole on cure rates after radioiodine treatment for Graves' hyperthyroidism: a randomized clinical trial. AB - Forty-two newly diagnosed patients with Graves' hyperthyroidism were randomly assigned to receive 131I therapy after pretreatment with methimazole (21) or beta blocker alone (21) and prospectively evaluated, to determine possible effects of methimazole on 131I treatment outcome. After randomization, 8 patients were excluded from the study (5 from pretreatment group and 3 from nonpretreatment group). Radioactive iodine (baseline dose 15 mCi, adjusted for goiter size and/or 131I uptake) was administered after pretreatment with methimazole (30 mg initial dose for at least 2 months and stopped 6 days before treatment) and beta-blocker or pretreatment with beta-blocker alone (atenolol 50-100 mg/d). All but one patient in each group became hypothyroid. A similar length of time was required by both groups to achieve hypothyroidism (112 days, [95% confidence interval [CI] = 28 to 196 days) in the pretreated group and 106 days, [95% CI = 45 to 167 days] in nonpretreated patients). Free thyroxine (T4) normalized 44 +/- 39 days after therapy in the nonpretreated group and 35 +/- 30 days in the pretreated group (p = 0.57) and decreased to subnormal levels 80 +/- 70 days in nonpretreated and 65 +/- 32 days in pretreated patients (p = 0.46). We conclude that pretreating patients with methimazole before radioactive iodine therapy does not interfere with the final outcome. Similar cure rates and time required to achieve hypothyroidism after radioiodine were observed when patients were pretreated with methimazole compared to nonpretreated patients. PMID- 11916283 TI - Temporal trends for thyroid carcinoma in Australia: an increasing incidence of papillary thyroid carcinoma (1982-1997). AB - The incidence of papillary thyroid carcinoma (PTC) has increased in many countries over the past 60 years. In Tasmania (an island state of the Australian Commonwealth with a well-documented history of iodine deficiency, subsequent supplementation and more recently a return to deficiency), there has been a fourfold rise in PTC incidence over the past two decades. The aim of this study was to evaluate thyroid carcinoma (TC) incidence trends in Australia, with particular reference to the roles of iodine nutrition, ionizing radiation, and ascertainment bias. Data from Australia's regional cancer registries were used to determine national TC incidence and mortality trends for the period 1982-1997. There were 9,053 new diagnoses of TC. Papillary, follicular, medullary, anaplastic, and "other diagnoses," accounted for 65.8%, 17.8%, 4.6%, 1.3%, and 10.5% of registered cases, respectively. TC incidence rates increased by 6.7% per year for females and 4.4% per year for males between 1982-1997 (p < 0.001). The increase was primarily because of a 10.7% per year and 8.3% per year rise in PTC incidence for females and males respectively (p < 0.001). The increase in PTC incidence was most significant for the population residing on Australia's eastern seaboard. The greatest relative and absolute increase in PTC (24.7% per year) occurred in Tasmania. These findings are consistent with a true increase in underlying PTC incidence rates. A link between current incidence trends and past iodine deficiency/radioiodine exposure during the 1950s and early 1960s is possible. PMID- 11916284 TI - Assessment of thyroid function in two hundred patients with beta-thalassemia major. AB - Despite improved hematologic care, multiendocrine dysfunction is a common complication of homozygous transfusion-dependent beta-thalassemia. In this study our goal was to estimate the prevalence of thyroid dysfunction in a large homogenous group of thalassemic patients. Two hundred patients with beta thalassemia major (100 males and 100 females; mean age, 23.2 +/- 6.7 years; age range 11-43 years), regularly transfused and desferioxamine chelated, were randomly selected from a pool of approximately 800 patients with beta-thalassemia followed in our department. Thyroid function and iron-load status were evaluated by measurements of free thyroxine (FT4), free triiodothyronine (FT3), thyrotropin (TSH), and serum ferritin levels. Of the subgroup of patients who proved to have normal thyroid hormone values, 26 (12 males, 14 females; mean age, 23.6 +/- 6.8 years; age range, 15-36 years) were randomly selected and underwent a standard TRH stimulation test. Thyroid dysfunction was defined as follows: overt hypothyroidism: low FT4 and/or FT3, increased TSH levels; subclinical hypothyroidism: normal FT4, FT3, increased TSH levels; exaggerated TSH response: normal FT4, FT3, normal basal TSH, deltaTSH > or = 21 microIU/mL (TSH levels measured prior and 30 minutes after intravenous TRH administration). Normal thyroid hormone values were found in 167 (83.5%) of the 200 patients studied. Eight (4%) of the remaining patients had overt hypothyroidisim, and 25 (12.5%) had subclinical hypothyroidism. Exaggerated TSH response to TRH was revealed in 7 of the 26 patients with normal hormone values tested (26.9%). Antithyroglobulin and anti-thyroid peroxidase (TPO) antibody titers were negative in 191 patients (95.5%). Mean ferritin levels in hypothyroid and euthyroid patients were 2707.66 +/- 1990.5 mg/L and 2902.9 +/- 1997.3 mg/L, respectively, (p = 0.61), indicating no correlation between ferritin levels and thyroid functional status. Mean ferritin levels in the patients who responded normally to TRH stimulation and in those who overresponded, were 2,586 +/- 1791 mg/L and 3,228 +/- 2473 mg/L, respectively (p = 0.46; NS). Thyroid failure is a rather rare endocrine complication in patients with beta-thalassemic from Greece. In our series, no case of central hypothyroidism was observed. No correlation was found between thyroid functional status and ferritin plasma levels. Approximately 1 of 5 beta thalassemic patients with normal thyroid hormone values showed an exaggerated TSH response to TRH test. It is to be investigated how many of these patients will establish overt or subclinical hypothyroidism in the future. PMID- 11916286 TI - Thyroid cancer in Vaud, Switzerland: an update. AB - We reviewed the descriptive epidemiology of thyroid cancer using data from the Cancer Registry of the Canton of Vaud, Switzerland, a consistently well surveilled population with relatively high rates of the disease, between 1974 and 1998, on the basis of a total of 596 registered cases. Overall thyroid cancer incidence tended to increase moderately in both genders over the 25-year period considered, to reach rates of 5.4 and 2.0 per 100,000 (world standard) in females and males, respectively. There were also changes in histologic classification, with some increases in papillary neoplasms and corresponding decreases in other and unclassified ones. Ten-year relative survival rates for cases diagnosed in 1988-1993 were 94% for papillary cancer in females and 69% in males, and 59% for follicular cancer in females. Corresponding figures were 12% for undifferentiated, and 31% for other and unspecified neoplasms in both sexes combined. Multivariate analysis confirmed the unfavorable influence of male gender (hazard ratio [HR] = 1.9), elderly age (HR = 16.5 for age > or = 65 vs. < 45 years) and undifferentiated histotype (HR = 3.5 vs. papillary) on the long term prognosis of thyroid cancer, and showed no consistent evidence of appreciably improved prognosis over more recent calendar periods. PMID- 11916285 TI - Implication of 2-18fluor-2-deoxyglucose positron emission tomography in the follow-up of Hurthle cell thyroid cancer. AB - The aim of this retrospective study was to investigate the value of positron emission tomography (PET) with 2-18fluor-2-deoxy-glucose (FDG) in the follow-up of Hurthle cell thyroid cancer (HTC), a rare variant of thyroid malignancies. FDG PET studies were performed in 17 patients with HTC. In subgroup A (n = 13) PET was initiated because of an elevated thyroglobulin (Tg) level whereas in subgroup B (n = 4) the study was performed to evaluate suspect findings of morphologic imaging while Tg remained undetectable. Pathologically increased FDG uptake was found in all patients of subgroup A. In 10 studies, PET results were proven as true-positive either by surgery or by morphologic imaging. One study was false positive. Final evaluation was not possible in two cases. In subgroup B, PET was true-negative in three and false-positive in one patient. For the detection of recurrent HTC by means of FDG-PET a meta-analysis including data of a multicenter study revealed an overall sensitivity of 92%, a specificity of 80%, a positive predictive value of 92%, and a negative predictive value of 80% while the accuracy was 89%. This study supports the efficiency of FDG-PET in the follow-up of HTC. PMID- 11916287 TI - Diagnostic fine-needle aspiration cytology and immunocytochemistry analysis of a primary thyroid lymphoma presenting as an anatomic emergency. AB - The case of a 66-year-old woman with rapidly progressive respiratory distress caused by a massive anterior neck mass with tracheal compression is presented. Within 24 hours, fine-needle aspiration cytology (FNAC) and immunocytochemistry provided a diagnosis of high-grade B-cell lymphoma and the opportunity to institute a chemotherapeutic regimen resulting in a rapid volume reduction and airway expansion. One year after combined modality treatment the patient was in complete remission (with an estimated thyroid volume of 4 cm3). This case report illustrates the advantages of FNAC and immunocytochemistry in the diagnosis of thyroid lymphoma. PMID- 11916288 TI - Acute suppurative thyroiditis caused by an infected piriform sinus fistula with thyrotoxicosis. AB - We report herein an unusual case of thyrotoxicosis caused by acute suppurative thyroiditis (AST) infected through a piriform sinus fistula (PSF). A 28-year-old man presented with pain over the thyroid gland and elevated serum thyroid hormone levels, a picture similar to subacute thyroiditis. A fine-needle aspiration biopsy from the left lobe showed neutrophil infiltration, and culture from the aspirate grew anaerobic peptostreptococcus. A neck computed tomography (CT) scan showed an abscess in the thyroid gland, and barium swallow revealed the presence of PSF. Appropriate antibiotic treatment ameliorated his symptoms of infection, followed by normalization of thyroid function. Three months later, he underwent fistulectomy and partial left lobectomy. The end of the PSF track was found in the left thyroid lobe. Thus infection of the thyroid gland through the infected PSF was likely the cause of supprative thyroiditis. The unusual clinical features of AST in this patient include the presence of severe thyrotoxicosis, relatively late onset (28-years-old) of infection despite the presence of congenital PSF, and the lack of acute inflammatory signs on the overlying skin of the thyroid gland. It is important to recognize this type of AST, since fistulectomy is required to prevent recurrent AST. PMID- 11916290 TI - MHC class II expression in Graves' disease. PMID- 11916289 TI - Thymus gland uptake of radioactive iodine. PMID- 11916291 TI - Making thyroid simple--avoid hyper and hypo! PMID- 11916292 TI - Tie-2 and angiopoietin-1 expression in human thyroid tumors. AB - Tie-2 is an endothelial cell-specific receptor tyrosine kinase involved in vascular maturation and remodeling. Although its expression is considered to be restricted to vascular endothelial cells and hematopoietic progenitors, our immunohistochemical and in situ hybridization studies showed that Tie-2 and its ligand, angiopoietin (Ang)-l were expressed not only in benign and malignant human thyroid tumor cells but also in hyperplastic regions of adenomatous goiter. To confirm the expression in these tissues further, we used a laser capture microdissection system to isolate epithelial tumor cells from tissue specimens selectively, and demonstrated the expression of Tie-2 and Ang-1 mRNAs in tumor cells by RT-PCR analysis. Furthermore, Tie-2 and Ang-1 mRNAs and proteins were also detected in rat thyroid cell lines, FRTL-5 and PCCL-3. Our results suggest that Ang-1/Tie-2 signaling may be involved in the proliferation of thyroid epithelial cells. PMID- 11916293 TI - Distance perception across spatial discontinuities. AB - We investigated the use of nested contact relations in perceiving the relative distance of locations on discontinuous surfaces. Observers viewed computer generated displays under monocular static conditions and adjusted a marker to match the perceived distance of a cube. The marker and cube were raised above the ground by two different platforms separated by a gap. The relative heights and distances of the platforms were varied. We found the following: (1) When spatially discontinuous surfaces are coplanar, locations of objects resting on these surfaces appear to be compared directly, bypassing relations with the underlying ground plane. (2) Spatial displacement between the platforms produces a bias, in the direction of the displacement, in the perceived relative locations of objects resting on the platforms. This suggests that local spatial relations between objects and their platforms are only partially integrated with more global spatial relations between the discontinuous surfaces of the platforms. PMID- 11916294 TI - Identification of scanned and static tactile patterns. AB - Most of the studies in which the interactions between target and nontarget spatial patterns have been examined have tested patterns that are generated statically. Static patterns are those in which all the elements of the pattern are presented at the same time and at a fixed location on the skin; however, most tactile information comes to the skin by means of patterns' being scanned across the surface of the skin. In the present study, the interactions between target and nontarget patterns were measured for patterns generated in both the static and the scanned modes. Nontarget patterns often interfere with the perception of target patterns. Using patterns generated in the static mode, previous studies have identified two factors that produce interference in pattern identification: response competition and masking. Masking, in turn, appears to be the result of temporal integration of the target and nontarget patterns, as well as the displacement of target features. In the present study, these factors were examined for patterns generated in both static and scanned modes. Regardless of the mode in which the patterns were generated, similar functions were obtained relating identification performance to the temporal separation between the target and the nontarget patterns. Although statically generated patterns are more easily identified than scanned patterns, particularly at brief durations, mechanisms such as response competition, temporal integration, and the displacement of target features appear to be factors that affect scanned patterns to nearly the same degree as static patterns. PMID- 11916296 TI - Perception of vocal effort and distance from the speaker on the basis of vowel utterances. AB - The sound pressure level of vowels reflects several nonlinguistic and linguistic factors: distance from the speaker, vocal effort, and vowel quality. Increased vocal effort also involves the emphasis of higher frequency components and increases in F0 and F1. This should allow listeners to distinguish it from decreased distance, which does not have these additional effects. It is shown that listeners succeed in doing so on the basis of single vowels if phonated, but not if whispered, and that they compensate for most of the between-vowel variation in level. The results obtained when listeners had to estimate vocal effort as well as distance suggest that an analysis of an utterance takes place at an early stage in auditory processing, before memories of episodes are stored. PMID- 11916295 TI - Timing relations in speech and the identification of voice-onset times: a stable perceptual boundary for voicing categories across speaking rates. AB - This study shows that the ratio of voice onset time (VOT) to syllable duration for /t/ and /d/ presents distributions with a stable boundary across speaking rates and that this boundary constitutes a perceptual criterion by which listeners judge the category affiliation of VOT. In Experiment 1, best-fit regression lines for VOT ratios of intervocalic /t/ and /d/ against speaking rate had zero slopes, and there was an inferable boundary between the distributions. In Experiment 2, listeners' identifications of syllable-initial stops conformed to this boundary ratio. In Experiment 3, VOT was held constant, while VOT ratios were altered by modifying the duration of the following vowel. As VOT ratios exceeded the boundary estimated from the data of Experiment 1, listeners' identifications shifted from /d/ to /t/. Timing relations in speech production can determine the identification of voicing categories across speaking rates. PMID- 11916297 TI - Binocular summation of chromatic changes as measured by visual reaction time. AB - We determined visual reaction times to monocular and binocular changes in the luminance of isochromatic stimuli and to monocular and binocular changes in the color of isoluminant stimuli. Two isoluminant color changes were tested: chromatic variations along the red-green axis of Boynton's (1986) two-stage color vision model and chromatic variations along the yellow-blue axis of the same model. The results indicate a greater degree of binocular summation for luminance change than for color change. This result was largely independent of the motor component of reaction time. PMID- 11916298 TI - Motor activation with and without inhibition: evidence for a threshold mechanism in motor control. AB - Masked primes presented prior to a target can trigger response activation processes that may later be subject to inhibition. Evidence for response inhibition has previously been obtained with primes presented at fixation, but not with primes presented in the periphery of the visual field. It is argued that this central-peripheral asymmetry reflects a threshold mechanism in motor control. Foveal masked primes give rise to stronger perceptual representations than do peripheral primes, resulting in stronger response activations. Strong response activations are actively inhibited, whereas weaker activations remain below a hypothetical inhibition threshold. Evidence in favor of this hypothesis is obtained in four experiments that manipulated the perceptual strength of foveal and peripheral primes. Experiments 1 and 2 demonstrate that when the perceptual strength of peripheral primes is gradually increased by delaying mask onset, positive prime-target compatibility effects (reflecting the absence of response inhibition) turn into negative effects (indicating the presence of response inhibition). Experiments 3 and 4 demonstrate that when the perceptual strength of foveal primes is gradually decreased by degradation, negative compatibility effects turn into positive effects. The results are discussed in terms of a simple functional model of early motor control. PMID- 11916299 TI - Trajectory forms as a source of information about events. AB - The ability to use trajectory forms as visual information about events was tested. A trajectory form is defined as the variation in velocity along a path of motion. In Experiment 1, we tested the ability to detect trajectory form differences between simulations of a freely swinging pendulum and a hand-moved pendulum. The trajectory form of the freely swinging pendulum was symmetric around the mid-point, whereas the hand moved was not. In Experiment 2, we isolated trajectory form information by varying the amplitudes of events while holding their periods constant. Straight path versions of the harmonic events from Experiment 1 were tested. In Experiment 3, we tested sensitivity to symmetrical peakening or flattening of trajectory forms. Participants detected small differences in all three experiments. In Experiment 4, we tested the ability to identify specific events based only on differences in trajectory forms. Participants were able to identify four different events. We investigated properties of trajectory forms that might potentially be detected and used as information, and we found that the curvature yielded good results. PMID- 11916300 TI - Perceptual latencies to discriminate surface orientation in stereopsis. AB - The difference in sensitivity to stereoscopic surfaces oriented horizontally or vertically (the stereoscopic orientation anisotropy) can be redescribed as a difference in sensitivity to shear or compression transformations that relate the binocular images. The present experiment was designed to test this by dissociating the image transformation from the orientation of the surface. Surfaces were presented in isolation or in the presence of a surrounding frame that formed step and gradient discontinuities in the disparity field. Without discontinuities, observers required considerably more time to discriminate between surfaces differing in compression than between those differing in shear, irrespective of surface orientation. Disparity discontinuities facilitated the perception of the disparity gradients; minimum stimulus durations were reduced by over an order of magnitude when the reference frame was present. These results support the hypothesis that the disparity field is decomposed into different primitives during the recovery of depth and surface structure. PMID- 11916301 TI - Object-based attention: sensory modulation or priority setting? AB - The detection of an invalidly cued target is faster when it appears within a cued object than when it appears in an uncued object equally distant from the cued location; this is a manifestation of object based attention. Five experiments are reported in which it was investigated whether early sensory enhancement (in which attention "spreads" within an attended object but stops at its borders) or a later attentional prioritization mechanism best accounts for these effects. In Experiments 1-4, subjects identified a centrally located target with a buttonpress while attempting to ignore flanking distractors that were mapped to either a compatible or an incompatible response. The flankers appeared either within the object occupied by the target or in a different object but at the same distance from the target. The well-known effect of distance between the target and the flankers on the magnitude of the compatibility effect was replicated. However, whether the target and the flankers were in the same or different objects had no effect on the magnitude of the compatibility effect. In Experiment 5, when attention could not be narrowly focused in advance, object-based modulation of the flanker effect was observed. These results suggest that object based selection may reflect an object-specific attentional prioritization strategy, rather than object-based attentional modulation of an early sensory representation. PMID- 11916302 TI - Object-based selection: the role of attentional shifts. AB - The objective of this paper was to investigate under what conditions object-based effects are observed. Recently, Watson and Kramer (1999) used a divided-attention task and showed that unless top-down factors induce a bias toward selection at a higher level, object-based effects are obtained when same-object targets belong to the same uniformly connected (single-UC) region, but not when they belong to different single-UC regions grouped into a higher order object (grouped-UC regions). We refine this claim by proposing that a critical factor in determining whether or not object-based effects with grouped-UC regions are observed is the need to shift attention. The results of four experiments support this hypothesis. Stimuli and displays were similar to those used by Egly, Driver, and Rafal (1994). Subjects had to make size judgments. Using different paradigms, we obtained object-based effects when the task required shifts of attention (spatial cuing, same vs. different judgment with asynchronous target onsets), but not when attention remained either broadly distributed (same vs. different judgment with simultaneous targets) or tightly focused (response competition paradigm). PMID- 11916303 TI - Modulations of the processing of line discontinuities under selective attention conditions? AB - We examined whether the processing of discontinuities involved in figure-ground segmentation, like line ends, can be modulated under selective attention conditions. Subjects decided whether a gap in collinear or parallel lines was located to the right or left. Two stimuli were displayed in immediate succession. When the gaps were on the same side, reaction times (RTs) for the second stimulus increased when collinear lines followed parallel lines, or the reverse, but only when the two stimuli shared the same orientation and location. The effect did not depend on the global form of the stimuli or on the relative orientation of the gaps. A frame drawn around collinear elements affected the results, suggesting a crucial role of the "amodal" orthogonal lines produced when line ends are aligned. Including several gaps in the first stimulus also eliminated RT variations. By contrast, RT variations remained stable across several experimental blocks and were significant for interstimulus intervals from 50 to 600 msec between the two stimuli. These results are interpreted in terms of a modulation of the processing of line ends or the production of amodal lines, arising when attention is selectively drawn to a gap. PMID- 11916304 TI - Decision strategies in the ABX (matching-to-sample) psychophysical task. AB - The ABX psychophysical task is very useful when the dimensions along which the stimulus varies cannot be readily described. However, there are a few problems associated with its use. First, the task has often been used with slight procedural variations, which hinders the extraction of a bias-free index of performance. Second, the most common index of sensitivity derived from this task is the ubiquitous proportion correct. Third, some researchers undervalue the importance that the decision strategy adopted by the observer has on estimates of sensitivity. We describe a standard methodology for the ABX task and outline models of performance based on two decision strategies. These models can be used to extract the detection-theoretic index of performance, d', from ABX data. The results of three ABX discrimination experiments, in which circle size, number value, and tone amplitude are used, are reported. Analysis of the decision spaces derived from observer responses indicates that these observers adopted a suboptimal difference decision strategy, even after they had received considerable practice with feedback. This suggests that, given no prior knowledge of the nature of the decision strategy adopted by an observer, the difference model of ABX discrimination could be a good choice when observer performance is estimated. PMID- 11916305 TI - Effective tumor immunotherapy: start the engine, release the brakes, step on the gas pedal,...and get ready to face autoimmunity. AB - Cellular immune responses can destroy cancer cells, achieving the cure of experimental malignancies. An expanding wealth of knowledge on the molecular basis of how to prime and amplify a T cell response has fueled a number of strategies successful at treating established tumors (rather than merely preventing tumor grafting). The most efficacious approaches operate at different stages, including: 1) priming the immune response using tumor antigen-expressing dendritic cells or tumor cells transfected with genes that render them immunogenic, 2) sustaining and amplifying immunity using agonistic monoclonal antibodies against costimulatory molecules or immune-potentiating cytokines, and 3) eliminating mechanisms that self-regulate the strength of the immune response, such as inhibitory receptors or regulatory T cells. A rational combination of such approaches holds great hope for cumulative and synergistic effects, but there is also evidence that they can open the flood-gates for unwanted inflammatory reactions. The next decade can be envisioned as the time when the first reproducibly efficacious combination regimes for cancer immunotherapy will become available and widely used in the clinic, as clinicians learn the best strategies and try to harness their potentially damaging effects. PMID- 11916306 TI - Caspases--their role in apoptosis and other physiological processes as revealed by knock-out studies. AB - Caspases are crucial mediators of apoptosis, a form of physiological cell death. Their activation is carefully controlled by a philogenetically conserved death program, which is indispensable for the homeostasis and development of higher organisms. Dysregulation of apoptosis contributes to the pathogenesis of many human diseases. As effectors of the apoptotic machinery, caspases are considered potential therapeutic targets. In vitro studies have demonstrated the requirement of caspase activity for both the triggering phase as well as the execution of apoptosis, thus providing a molecular base for the fine-tuning of this process by pharmacological agents. The precise roles of the individual caspases in vivo and their functional relation to each other have been best demonstrated in genetically modified animals. The generation of single caspase-deficient mice have confirmed most of the data obtained in vitro and exposed some new aspects previously undetected in the cell culture system. Interestingly, inactivation of many caspases revealed not only their expected participation in apoptotic events as well as in the maturation of cytokines, but also provided hints about the role of at least some caspases in cell differentiation and stimulatory responses. In this review we will discuss what these studies have unveiled about the role of individual caspases in development, apoptosis, and inflammation, with particular focus on their role beyond the apoptotic process. PMID- 11916307 TI - Emerging concepts in the molecular pathogenesis of systemic lupus erythematosus. AB - Systemic lupus erythematosus is a prototypic autoimmune disease that afflicts predominantly women during their child-bearing age. The disease is characterized by the production of autoantibodies and immune complexes in association with a diverse array of clinical manifestations. Investigation into the etiopathogenesis has been directed at identifying the genes that provide susceptibility to the disease, the complex cellular and cytokine aberrations and the biochemical abnormalities that are responsible for them. Understanding the immune cell signaling and gene transcription abnormalities will help us tailor new strategies for efficient biotherapy of the disease. PMID- 11916308 TI - Interferon tau and its immunobiological role in ruminant reproduction. AB - Interferon tau (IFN-tau) is an key cytokine in maintaining pregnancy in ruminants. It is produced by the ruminant conceptus around the time of implantation. IFN-tau belongs to the type I interferon family but, unlike the other members of this group, it is not virus inducible and its expression is temporal and restricted to the trophoblast cells of the ruminant conceptus. The main target of the paracrine action of this cytokine is the endometrium. It changes the prostaglandin metabolism and secretory function of the cells by upregulating the secretion of several proteins. It also presents immunomodulatory action towards leukocytes by changing their proliferative responses and cytokine production. This cytokine activity in reproductive biology and immunology has been intensively explored for the last ten years. It has been regarded as a potential tool in improving the performance and biotechnological processes in ruminant reproduction. Additionally, its high antiviral potency and low cytotoxicity in comparison with IFN-alpha has placed this cytokine in the group of possible therapeutics in human and animal medicine. PMID- 11916309 TI - The role of the innate and adaptive immune responses in Acanthamoeba keratitis. AB - Infections of the corneal surface are an important cause of blindness. Protozoal, viral, bacterial, and helminthic infections of the cornea account for up to 9 million cases of corneal blindness. Free-living amoebae of the genus Acanthamoeba produce a progressive infection of the cornea called Acanthamoeba keratitis. Disease is usually transmitted by Acanthamoeba trophozoites bound to soft contact lenses. Infection of the cornea is initiated when the parasite binds to the corneal epithelial surface. Recrudescence can occur and suggests that the adaptive immune response is not aroused by corneal Acanthamoeba infections. Systemic immunization with Acanthamoeba antigens elicits robust Th1 cell-mediated immunity and serum IgG antibody, yet fails to prevent the development of Acanthamoeba keratitis. However, immunization via mucosal surfaces induces anti Acanthamoeba IgA antibodies in the tears and provides solid protection against the development of Acanthamoeba keratitis. Unlike other immune effector mechanisms that rely on cytolysis, inflammation, release of toxic molecules, or the induction of host cell death, the adaptive immune apparatus prevents Acanthamoeba infections of the cornea by simply preventing the attachment of the parasite to the epithelial surface. The beauty of this mechanism lies in its exquisite simplicity and efficacy. PMID- 11916310 TI - Thermal glycation of proteins by D-glucose and D-fructose. AB - The dry thermal glycation method was used to conjugate D-glucose and D-fructose with bovine serum albumin. Reactions were conducted at 50-104 degrees C for 30 min. Depending on temperature, different levels of substitution were achieved. In the case of D-glucose, average substitution levels of up to 53 mol glucose/mol bovine serum albumin were obtained. D-fructose turned out to be less reactive than D-glucose. The pH of the reaction mixture was also found to affect the efficiency of the glycation reaction. The levels of substitution were estimated using matrix-assisted laser desorption/ionisation time-of-flight mass spectrometry and SDS-polyacrylamide gel electrophoresis. Glycated molecules, and a process of glycation itself are implicated in diabetes complications. Hypothetically bovine spongiform encephelopathy (BSE) was also derived from glycation. PMID- 11916312 TI - Comparison of leukocyte populations from bronchoalveolar lavage and induced sputum in the evaluation of cellular composition and nitric oxide production in patients with bronchial asthma. AB - Bronchoalveolar lavage (BAL) or induced sputum (IS) techniques may provide leukocytes for the evaluation of airway inflammatory response in bronchial asthma. The aim of the present study was to compare features of leukocyte populations obtained by the two different methods regarding the cell types and their activity in patients with bronchial asthma. The nitric oxide (NO) level released from the cells was measured as a marker of their activity. Pulmonary leukocytes were obtained from the BAL and IS of 11 asthmatic patients in stable condition at the time of the study. The BAL and IS leukocyte populations varied in cell count and NO production. Macrophages were the predominant leukocyte population in BAL (median (Me) = 83.0%, range 67.9-88.4%), whereas sputum sediments were found to consist mainly of neutrophils (Me = 55.7%, range 29.0 64.9%). The IS leukocytes released much more NO (p = 0.0022) than the BAL leukocytes. In spite of these quantitative differences, a similar pattern of NO production was observed in BAL and in IS cells. Both BAL and IS leukocyte populations produced almost the same amounts of NO before and after lipopolysaccharide stimulation (p = 0.9063, p = 0.4801, respectively). Furthermore, a slight positive correlation Spearman's rank (RS) = 0.5578, p = 0.0594) was noticed between the neutrophil percentages and NO levels produced by BAL cells, whereas in IS a statistically significant correlation between the percentage of neutrophils and the levels of NO (RS = 0.6643, p = 0.0184) was observed. In conclusion, the BAL and IS leukocyte populations are different in cell type, their size and activity. Depending on the asthma severity and the type of cells needed in a study, either BAL or IS specimens may be chosen as a source of pulmonary leukocytes. The use of IS as a noninvasive technique is supposed to be potential value particularly in the study of the airway inflammatory response mediated mainly by neutrophils, i.e. during and/or after exacerbation of the disease. Based on our results, a possible contribution of neutrophils in the production of NO in the airways of asthmatic patients can be proposed apart from other cells such as macrophages. PMID- 11916311 TI - Glycoforms of six serum glycoproteins in a patient with congenital disorder of glycosylation type I. AB - In this paper the occurrence and relative content of defectively glycosylated serum glycoforms in transferrin (Tf), alpha1-acid glycoprotein (AGP), haptoglobin (Hp), alpha1-antitrypsin (alpha1-AT), alpha2-macroglobulin (alpha2-MG) and ceruloplasmin (Cpl) in the serum of a patient with congenital disorder of glycosylation type I are reported. Blood samples were taken when the patient was 14 years old and then after a one-year interval. The patterns of glycoforms in both samples were compared. In 4 out of 6 examined glycoproteins, glycoforms lacking one and two oligosaccharide chains occurred. "Underglycosylated" glycoforms of alpha2-MG and Cpl were not clearly detectable. Tf was shown to be affected with this defect to a higher extent than other glycoproteins, containing only 30% properly glycosylated molecules and also as much as 30% of the molecules lacking two glycan units. In Hp and alpha1-AT the proportions of properly and defectively glycosylated forms were similar. This properly glycosylated form comprised 47% of the Hp and 51-55% of the alpha1-AT molecules. As in AGP and Tf, about 30% the of molecules lacked one glycan unit. Twenty-one percent of the Hp molecules were devoid of two glycans, and this amount slightly increased in the course of the year. In alpha1-AT, 19 and 17% of the molecules lacked two glycans in both samples, respectively. Only in AGP we did find a substantial difference between the two blood samples. In the course of the year, the amount of the form lacking 2-chains decreased from 12 to 3%, resulting in a simultaneous increase in the forms lacking one chain and the properly glycosylated. Our work also indicates, that applying a simple method of biochemical analysis such as SDS PAGE/Western-blotting could be helpful in preliminary diagnosis and could improve the identification of congenital disorders of glycosylation. PMID- 11916313 TI - Modelling the phenylalanine blood level response during treatment of phenylketonuria. AB - A vast body of phenylketonuria (PKU) patient monitoring data is deposited in clinical files and, after having served the actual needs, has remained there largely unused. We propose a kinetic model that will allow continued analysis of such data for further elucidation of the patient's metabolic phenotype and phenylalanine (Phe) disposal characteristics. Our PKU model of a single compartment with the input of alimentary Phe and two outputs--(1) first-order Phe conversion to tyrosine and acidic metabolites, and (2) zero-order Phe usage for net protein synthesis--has been developed with the graphics-oriented ModelMaker (then Cherwell Scientific Ltd, Oxford, UK) software package. The corresponding differential and integrated rate equations are presented to enable transfer of the model to equation-oriented simulation packages. The model offers a possible explanation for discrepancies in some genotype-phenotype data. PMID- 11916314 TI - Treatment and outcome of Taiwanese patients with 6-pyruvoyltetrahydropterin synthase gene mutations. AB - Ten cases of tetrahydrobiopterin (BH4) deficiency were identified in 1,337,490 newborns screened in a Chinese population in Taiwan. The high incidence of BH4 deficiency in the Taiwanese population may be explained by a founder effect, since all of the patients revealed 6-pyruvoyltetrahydropterin synthase gene mutations, and grouping N52S and P87S mutations together constituted 88.9% of the disease alleles. BH4 supplementation with restriction of high-protein foods gave control of plasma phenylalanine within normal range, and levodopa itself prevented seizure. However, the average intelligence quotient (IQ) score of these patients was only 76 +/- 14 (56-98). Statistically, the age of starting medication, including 5-hydroxytryptophan (5-HTP), was inversely correlated to IQ scores of these patients. We suggest the combination of BH4, levodopa and 5-HTP as the standard protocol to commence the treatment of BH4 deficiency as early as possible, although prenatal brain damage could have existed. PMID- 11916315 TI - Outcome of tyrosinaemia type III. AB - Tyrosinaemia type III is a rare disorder caused by a deficiency of 4 hydroxyphenylpyruvate dioxygenase, the second enzyme in the catabolic pathway of tyrosine. The majority of the nine previously reported patients have presented with neurological symptoms after the neonatal period, while others detected by neonatal screening have been asymptomatic. All have had normal liver and renal function and none has skin or eye abnormalities. A further four patients with tyrosinaemia type III are described. It is not clear whether a strict low tyrosine diet alters the natural history of tyrosinaemia type III, although there remains a suspicion that treatment may be important, at least in infancy. PMID- 11916316 TI - Impact of new mutations in the methylenetetrahydrofolate reductase gene assessed on biochemical phenotypes: a familial study. AB - Methylenetetrahydrofolate reductase (MTHFR) deficiency was identified in two out of four children born from nonconsanguineous parents. One of the affected children exhibited some clinical findings suggesting cystathionine beta-synthase deficiency; MTHFR activity was extremely reduced. In addition, hyperhomocysteinaemia, hypomethioninaemia, low total folate, especially methylfolate in red blood cells, and a reduced methylfolate/total folate ratio were found. Two mutations not yet reported, one on exon 1 of the gene changing an arginine to stop codon and one other on exon 9 changing an arginine to tryptophan were identified in both children in the compound heterozygous state associated with a common polymorphism, 1298A>C, also in the heterozygous state. The mother, homozygous for the mutation on exon 9 and for the polymorphism 1298A>C on exon 7, was clinically and biochemically normal, with normal folate status, mainly methylfolate levels in red blood cells, although MTHFR activity was moderately decreased. The father, heterozygous for the transition arginine to stop codon and for the common polymorphism 677C>T on exon 4, exhibited major biochemical abnormalities, hyperhomocysteinaemia and low methylfolate levels in red blood cells, but was clinically normal. The unaffected children had a biochemical pattern close to that of their mother and were heterozygous for the mutation on exon 9 and also for the two common polymorphisms, 677C>T and 1298A>C. In the affected children, some biochemical abnormalities, including folate status, especially methylfolate levels, were improved with treatment combining methyltetrahydrofolic acid, hydroxocobalamin, pyridoxine and betaine; however, homocysteine concentrations remained high and methionine concentrations were lowered. The father was treated with folic acid, which partially improved biochemical abnormalities. The impact of these mutations is discussed. PMID- 11916317 TI - Prolidase deficiency diagnosed by 1H NMR spectroscopy of urine. AB - Three urine samples from two prolidase-deficient patients were analysed using 1H NMR spectroscopy. One-dimensional 1H NMR spectra showed a characteristic pattern of overlapping resonances of the proline and hydroxyproline protons of the imidodipeptides. The model compounds Ala-Pro, Gly-Pro, Phe-Pro, Leu-Pro, Val-Pro, Gly-Hyp and Pro-Hyp were measured as well. The non-proline resonances of Val-Pro, Ala-Pro and Gly-Pro could be assigned in the urine spectra. These resonances could then be used for quantification of the corresponding imidodipeptids. The presence of Leu-Pro in the patients' urine was demonstrated by the results of COSY experiments. However, this imidodipeptide could not be quantified owing to overlap of the resonaces in the one-dimensional 1H NMR spectrum of the patients' urine. Phe-Pro, Pro-Hyp and Gly-Hyp could not be assigned in the spectrum of the patient's urine. The characteristic resonances in the urine from a prolidase deficient patient, i.e. Ala-Pro, Val-Pro, Gly-Pro, and resonances of the (hydroxy)proline part of the imidodipeptides can be used to diagnose this disease. PMID- 11916318 TI - Decreasing serum VLCFA levels in ageing X-ALD female carriers. AB - Adrenoleukodystrophy is an X-linked severe demyelinating disease with pathognomonic accumulation of saturated very long-chain fatty acids (VLCFA) in tissues and body fluids in affected males. The identification of women heterozygotes is also based on increased serum VLCFA concentrations. We describe the results of measuring serum VLCFA concentrations in 59 females of various ages with heightened risk of carrier status. In female carriers aged 22-50 years we found serum VLCFA concentrations in a range characteristic of heterozygotes; VLCFA levels were normal in female carriers aged 55-64 years. In women aged 37-50 years in whom repeat studies of VLCFA concentration were performed after 5 years, a reduction in VLCFA was observed. The results we obtained point to a reduction of serum VLCFA concentrations as X-ALD heterozygotes age. PMID- 11916319 TI - Hyperinsulinaemic hypoglycaemia--leading symptom in a patient with congenital disorder of glycosylation Ia (phosphomannomutase deficiency). AB - A male infant is described who presented with persistent hyperinsulinaemic hypoglycaemia, responding to diazoxide treatment. However, this therapy was discontinued because of seizures as a consequence of disturbed water and electrolyte balance. Glucose homeostasis could only be maintained by subtotal pancreatectomy, which was performed at 3 8/12 years of age. He developed a severe thrombosis, whereon a congenital disorder of glycosylation (CDG) was suspected. An abnormal transferrin isoelectric focusing pattern was found and the diagnosis of CDG Ia was confirmed by enzyme and molecular genetic analysis. This is the first patient with phosphomannomutase deficiency (McKusick 601785) described presenting with severe hyperinsulinaemic hypoglycaemia. PMID- 11916320 TI - Continuous glucose monitoring in children with glycogen storage disease type I. AB - Glycogen storage disease type I (GSD I) is characterized by impaired production of glucose from glycogenolysis and gluconeogenesis resulting in severe fasting hypoglycaemia. The aim of the present study was to examine the efficacy of a continuous subcutaneous glucose monitoring system (CGMS MiniMed), to determine the magnitude and significance of hypoglycaemia in GSD I and to evaluate the efficacy of its dietary treatment. Four children with GSD I were studied over a 72-h period. Results indicated that the values recorded with continuous subcutaneous glucose monitoring were highly correlated with paired blood glucose values measured by glucometer. Significant periods of asymptomatic hypoglycaemia were noted, especially during night-time. The study suggests that repeated continuous subcutaneous glucose monitoring may serve as a useful tool for the assessment of the long-term management of GSD I patients. PMID- 11916321 TI - Therapeutic trial in the first three Asian cases of ethylmalonic encephalopathy: response to riboflavin. AB - Three Korean girls with ethylmalonic encephalopathy, the first Asian cases, were identified. In all three cases, we observed slight improvement in motor functions, cognitive behaviours and chronic mucoid diarrhoea after treatment with riboflavin and/or coenzyme Q10 treatment. The precise pathogenesis of ethylmalonic encephalopathy has not been fully elucidated, but riboflavin treatment may be helpful. PMID- 11916322 TI - Severe hypoglycaemia in a patient with glycogen storage disease type III induced by infectious mononucleosis. AB - A 10-month-old girl with glycogen storage disease type III developed recurrent severe hypoglycaemia induced by infectious mononucleosis. Severe metabolic damage probably reflected a rapid breakdown of liver cells induced by the viral infection. PMID- 11916323 TI - Familial hypomagnesaemia with secondary hypocalcaemia: a new case that indicates autosomal recessive inheritance. AB - Male and female siblings, now aged 18 and 23 years respectively, with familial hypomagnesaemia and secondary hypocalcaemia provide further evidence that this is an autosomal recessive disorder and not X-linked as originally thought. PMID- 11916324 TI - Selective screening for inborn errors of metabolism: the primary care-based model in rural Crete. PMID- 11916325 TI - Glucose-6-phosphatase gene mutations in Turkish patients with glycogen storage disease type Ia. PMID- 11916327 TI - Liver transplantation in a case of argininaemia. PMID- 11916326 TI - Clinical and molecular findings in four new patients harbouring the mtDNA 8993T>C mutation. PMID- 11916328 TI - Diagnostic conceptualization of autism among Indian psychiatrists, psychologists, and pediatricians. AB - Diagnostic criteria for autism and background characteristics used by 937 Indian psychiatrists, psychologists and pediatricians were examined. Participants were asked to rate 18 behaviors as necessary for a diagnosis of autism, helpful but not necessary, or not helpful in a diagnosis of autism, and were asked to provide other information about their experiences with autism. Professionals' experience with diagnosing cases did not vary by profession and, in general, the three professions agreed about the characteristics most necessary for a diagnosis. However, within-group differences were found on the agreement over the usefulness of individual characteristics and amount of experience diagnosing cases as autistic. Comparisons with DSM-III and DSM-IV criteria suggest that Indian professionals may adhere to these systems. Conclusions about diagnosis in a cultural context and areas for future research are suggested. PMID- 11916329 TI - Coping by redefinition: cognitive appraisals in mothers of children with autism and children without autism. AB - To test a model of how mothers cope with the stresses of raising a child with autism (Tunali & Power, 1993), mothers of children with and without autism were interviewed. As predicted, mothers of autistic children: (1) placed less emphasis on career success and were more likely to believe that mothers of young children should not work outside of the home; (2) spent more leisure time with their extended family; (3) placed less emphasis on others' opinions of their child's behavior; (4) placed more emphasis on spousal support and parental roles in their discussions of marriage; (5) had more difficulty understanding their child's behavior; and (6) showed a marginally significant difference in their tolerance of ambiguity. Moreover, mothers of children with autism who showed these characteristics had the greatest life satisfaction overall. PMID- 11916330 TI - A descriptive study of hyperlexia in a clinically referred sample of children with developmental delays. AB - In this study, we evaluated the incidence of hyperlexia in a clinically referred sample of 80 children with developmental delays. Based on hypotheses previously formulated in the literature, the study investigated the frequency of hyperlexia among boys and girls, the incidence of hyperlexia in children with Pervasive Developmental Disorders (PDD)-spectrum compared with non-PDD diagnoses, the range of IQ and of various cognitive skills in children with and without hyperlexia, and the developmental outcomes of children with and without hyperlexia. The results revealed no significant differences in the frequency of hyperlexia in girls compared with boys. However, the frequency of hyperlexia was significantly elevated among children with PDD compared with children with non-PDD diagnoses. The range of IQ and other cognitive skills and the developmental outcomes of children with hyperlexia were comparable to those of children without hyperlexia. PMID- 11916331 TI - Molecular and cytogenetic analyses on Brazilian youths with pervasive developmental disorders. AB - The Pervasive Developmental Disorders (PDDs) constitute a group of behavioral and neurobiological impairment conditions whose main features are delayed communicative and cognitive development. Genetic factors are reportedly associated with PDDs and particular genetic abnormalities are frequently found in specific diagnostic subgroups such as the autism spectrum disorders. This study evaluated cytogenetic and molecular parameters in 30 youths with autism or other PDDs. The fragile X syndrome was the most common genetic abnormality detected, presented by 1 patient with autism and 1 patient with PPD not-otherwise specified (PPD-NOS). One girl with PDD-NOS was found to have tetrasomy for the 15q11-q13 region, and one patient with autism exhibited in 2/100 metaphases an inv(7)(p35q36), thus suggesting a mosaicism 46,XX/46,XX,inv(7)(p15q36) or representing a coincidental finding. The high frequency of chromosomopathies support the hypothesis that PDDs may develop as a consequence to chromosomal abnormalities and justify the cytogenetic and molecular assessment in all patients with PDDs for establishment of diagnosis. PMID- 11916332 TI - Random number generation in autism. AB - This study explored the ability of individuals with autism to generate a unique series of digits. Fourteen low-functioning individuals with autism, 14 intellectually disabled individuals, and 14 postgraduate university students generated a series of pseudo-random digits. Individuals with autism were more likely to repeat previous digits than were either of the control groups. The normal control group, however, was less likely to attempt cycling through all digits before repeating. Accordingly, low-functioning individuals with autism may exhibit a short-fall in response inhibition. This finding supports the executive dysfunction theory of autism. PMID- 11916333 TI - Asperger's disorder and mathematicians of genius. PMID- 11916334 TI - Legitimacy of comparing fragile X with autism questioned. PMID- 11916335 TI - Rulebound and nonrulebound tasks. PMID- 11916336 TI - Predicting outcome in autism. PMID- 11916337 TI - The risk of writing: putting your thoughts "on the line". PMID- 11916338 TI - Outcomes, structure, and process. PMID- 11916339 TI - Rural-urban partnering in continuing education. AB - BACKGROUND: One urban center provided an Advanced Cardiac Life Support (ACLS) course to a rural hospital using a minimum of on-site instructors. METHOD: Interactive television (ITV) conferencing was used to educate health care providers in a hospital more than 100 miles away. RESULTS: Nurses and physicians from a small rural hospital in northern Nevada were able to "attend" a full-scale ACLS course without having to leave their hospital, resulting in minimal cost to the agency. CONCLUSION: Rural and urban hospitals can partner to provide high quality, cost-effective education to nurses and other health care providers through the use of ITV conferencing. PMID- 11916340 TI - Publication syndicates: in support of nursing scholarship. AB - BACKGROUND: Within the university sector, one's corporate worth is, to a great extent, measured by research output, including refereed publications. Currently, only 7% of nurse academics publish each year. If nurses are to be competitive in the university arena and close the research-practice gap, they must be encouraged to publish. METHOD: This article examines publication rates within nursing, explores the role publication syndicates can play in supporting manuscript development, and offers a case study on the development of a publication syndicate within a School of Nursing at Griffith University, Australia. RESULTS: Syndicate members increased their publication rates two-fold, engaged in additional collaborative ventures, and demonstrated a renewed interest in writing for publication. DISCUSSION: Case study results confirmed that publication syndicates can decrease manuscript development time, increase the quality of work, influence productivity, and support collaborative faculty activities. PMID- 11916341 TI - The learning needs of nurses experiencing job change. AB - This study explored the perceived knowledge needs of nurses who experienced job change because of health care restructuring and whether different types of job change influenced learning needs. A questionnaire was distributed to nurses in two teaching hospitals. Results indicated that nurses perceived they needed new knowledge and upgraded clinical skills when experiencing job change. Their learning needs, however, differed with the type of job change experienced. Additionally, nurses felt their orientation only moderately prepared them for their new jobs. The findings suggest orientation programs need to be tailored to address the learning needs of nurses experiencing job change. PMID- 11916342 TI - Reflections on the changing learning needs of nurses: a challenge for nursing continuing educators. AB - Assessing the needs of learners is an essential prerequisite to planning effective nursing continuing education activities. Recorded differences over a 5 year period of gathering such assessments at one university-based continuing education division have relevance for nursing continuing educators and nursing leaders. A shift to more personal psychological interests of nurses may be reflective of the stressors of a changing system of health care delivery during this same time period. Nursing continuing educators are challenged to assist nurses to adapt successfully to these changes in the health care system and in the profession of nursing. PMID- 11916343 TI - Enhancing metacognition through the reflective use of self-regulated learning strategies. AB - BACKGROUND: An important concern of nursing practice and education is the difficulty new graduates experience while making the transition from graduate nurse to practicing nurse. METHOD: Using a comparative descriptive design, self regulated learning strategies were used to enhance metacognitive critical thinking abilities as 32 new graduate nurses reflected during 8-week preceptorship programs. RESULTS: Verbal protocol analysis revealed the majority of noun referents as metacognitive with thinking nouns increasing in rank from Week 1 to Week 8, present tense verbs were used most frequently with lower-level thinking phrases. Common themes in the narrative were knowledge observation, thinking strategies, judgments of self-improvement, judgments of competence, judgments of resources, self-reactions, and self-correction strategies. CONCLUSIONS: New graduate nurses have unique circumstances to overcome in making a transition to the workplace, and having self-regulatory skills would enable this process. The data suggest nursing education and practice consider self regulated learning prompts with new graduates to promote thinking strategies. PMID- 11916344 TI - New choices for continuing education: a statewide survey of the practices and preferences of nurse practitioners. AB - Technological innovations in the past decade have made possible several promising new modes for delivering continuing education (CE). Offering a wide variety of educational approaches is necessary to satisfy the different learning needs and preferences of program participants. Continuing education planners need to assess the preferences and practices of Advanced Practitioners of Nursing (APNs) when choosing the modes they will offer for delivering CE programs. A survey was conducted with the entire population of licensed APNs in Nevada to assess practices, preferences, and barriers to use of various CE delivery modes. In person conferences and live satellite conferences were the most frequently used methods. The top three preferences, in rank order, were in-person conference, print-based self-study, and interactive video conference. Live satellite conference was the least preferred method of earning CE credits. Computer-based modes of CE delivery, which include the Internet and CD-ROM, were among the least used. Findings from this study provide useful information for planners of CE programs for APNs. Data acquired in this study also address the dearth of information related to computer use by APNs for obtaining CE. PMID- 11916345 TI - Gastric cancer in sub-Saharan Africa. AB - Gastric cancer has a variable but generally low prevalence in black populations of sub-Saharan Africa, despite a high prevalence of Helicobacter pylori (the 'African enigma'). Evidence from Soweto indicates that the host response to H. pylori may be protective against a virulent organism and that, in most people, H. pylori does not lead to more serious sequelae. This suggests that there may be host protective/inhibitory factors present, which prevent the progression of H. pylori-induced chronic active gastritis to cancer. PMID- 11916346 TI - Carditis, intestinal metaplasia and adenocarcinoma of oesophagogastric junction. AB - Barrett's oesophagus is a precancerous condition in which the normal squamous epithelium is replaced by intestinal metaplasia (IM). IM can then progress through increasingly severe dysplasia to oesophageal adenocarcinoma (EAC). In the gastric cardia the normal gastric mucosa, when inflamed (carditis), can be replaced by IM and can then progress to gastric adenocarcinoma (GAC). The same histopathological sequence can take place on either side of the oesophagogastric junction. Since the location of that junction can be uncertain this can result in confused diagnosis between EAC and GAC. In this review, the diagnostic criteria, incidence and risk factors for Barrett's oesophagus and carditis are discussed, together with the factors determining the risk of progression to adenocarcinoma of the oesophagus or cardia. The risk factors include familial/genetic, environmental and dietary characteristics. Finally, these risk factors are discussed within the context of cancer prevention. PMID- 11916347 TI - The Malmo Diet and Cancer Study: representativity, cancer incidence and mortality in participants and non-participants. AB - In order to investigate potential selection bias in population-based cohort studies, participants (n = 28098) and non-participants (n = 40807) in the Malmo Diet and Cancer Study (MDCS) were compared with regard to cancer incidence and mortality. MDCS participants were also compared with participants in a mailed health survey with regard to subjective health, socio-demographic characteristics and lifestyle. Cancer incidence prior to recruitment was lower in non participants, Cox proportional hazards analysis yielded a relative risk (RR) with a 95% confidence interval of 0.95 (0.90-1.00), compared with participants. During recruitment, cancer incidence was higher in non-participants, RR: 1.08 (1.01 1.17). Mortality was higher in non-participants both during, 3.55 (3.13-4.03), and following the recruitment period, 2.21 (2.03-2.41). The proportion reporting good health was higher in the MDCS than in the mailed health survey (where 74.6% participated), but the socio-demographic structure was similar. We conclude that mortality is higher in non-participants than in participants during recruitment and follow-up. It is also suggested that non-participants may have a lower cancer incidence prior to recruitment but a higher incidence during the recruitment period. PMID- 11916348 TI - IQ (2-amino-3-methylimidazo[4,5-f]quinoline)-induced aberrant crypt foci and colorectal tumour development in rats fed two different carbohydrate diets. AB - In most aberrant crypt foci (ACF) and colorectal tumour studies, chemical carcinogens not normally found in food have been used as initiators. In the present study the food-related compound, IQ (2-amino-3-methylimidazo[4,5 f]quinoline), has been used. A diet high in refined carbohydrates has been associated with enhanced development of ACF and colorectal cancer in humans. The present study was designed as an integrated part of our earlier published ACF study and follows the animals until tumour development. The aim of the study was to investigate (1) the effect of a refined carbohydrate-rich diet on the development of IQ-induced ACF over time and (2) possible correlation between early and late ACF and/or colorectal tumour development. The study showed that a feeding regimen with continuous doses of 0.03% IQ in the diet for 14 weeks, followed by 32 weeks without IQ was able to induce tumours in the rat colon, liver, skin and Zymbal gland. The data demonstrate that a sucrose-rich diet enhance ACF development. A correlation between the outcome of early and late ACF was seen. However, as the tumour incidence of this study was very low it was not possible to obtain a meaningful correlation between ACF development and colorectal tumour incidence. PMID- 11916349 TI - In vitro and in vivo (SCID mice) effects of phytosterols on the growth and dissemination of human prostate cancer PC-3 cells. AB - The dietary effect of phytosterols (PS) versus cholesterol on the growth and metastasis of the PC-3 human prostate cancer cells in SCID mice was studied. Also, their direct effect on the growth and migration of these cells in vitro was analysed. In the in vivo experiment, SCID mice were fed a diet containing 2% of either PS mixture or cholesterol plus 0.2% cholic acid and implanted with 2 x 10(6) tumour cells per mouse. Tumour growth was monitored for 8 weeks post inoculation. Animals fed the PS diet had tumours 40-43% smaller than those fed the cholesterol diet. Furthermore, the number of mice with lymph node and lung metastasis was almost one-half that of the cholesterol-fed group. In the in vitro studies, both beta-sitosterol and campesterol inhibited the growth of PC-3 cells by 70% and 14%, respectively, while cholesterol supplementation increased the growth by 18% when compared with controls. PS inhibited the invasion of PC-3 cells into Matrigel-coated membranes by 78% while cholesterol increased it by 43% as compared with the cells in the control media. Migration of tumour cells through 8 microm pore membranes was reduced by 60-93% when the PC-3 cells were in PS media, as compared with a 67% increase after cholesterol supplementation. PS supplementation reduced the binding of PC-3 cells to laminin by 15-38% and fibronectin by 23% while cholesterol increased binding to type IV collagen by 36%. It was concluded that PS indirectly (in vivo as a dietary supplement) and directly (in tissue culture media) inhibited the growth and metastasis of PC-3 cells. beta-Sitosterol was more effective than campesterol in offering this protection in most of the parameters studied. PMID- 11916350 TI - Low-risk diet for colorectal cancer in Italy. AB - An innovative approach was used to define a low-risk diet for colorectal cancer from a multicentric case-control study of 1953 incident cases and 4154 hospital controls from Italy. A logistic regression model was fitted on the reported intake of five macronutrients, and the estimated coefficients were used to compute a diet-related logistic risk score (LRS). The mean of LRS within risk decile ranged from 0.89 to 1.86. Total energy intake and absolute consumption of each macronutrient increased with increasing LRS. In relative terms, however, starch intake showed an almost threefold increase across subsequent score levels, while a decline was observed for unsaturated fat, sugar and protein. Saturated fat consumption remained fairly stable in relative terms. When food groups were considered, bread and cereals dishes, cakes and desserts and refined sugar were positively associated, while the consumption of vegetables, fruit, fish, poultry and olive oils was inversely associated with LRS. PMID- 11916351 TI - Ionizing radiation, cellular telephones and the risk for brain tumours. AB - A case-control study on brain tumours included 233 patients aged 20-80 years and alive at the study time. They had histopathologically verified brain tumour and lived in the Uppsala-Orebro region (1994-1996) or the Stockholm region (1995 1996). Two matched controls to each case were selected from the Swedish Population Register. Two hundred and nine cases (90%) and 425 controls (91%) answered the questionnaire. Results are presented for the whole study group, as given here, and for malignant and benign tumours separately. For workers in the chemical industry the odds ratio (OR) was 4.10, 95% confidence interval (95% CI) 1.25-13.4 and laboratory workers OR 3.21, 95% CI 1.16-8.85. Radiotherapy of the head and neck region gave OR 3.61, 95% CI 0.65-19.9. Medical diagnostic X-ray of the same area yielded OR 1.64, 95% CI 1.04-2.58. Work as a physician gave OR 6.00, 95% CI 0.62-57.7. All three cases had worked with fluoroscopy. Ipsilateral (same side) use of a cellular telephone increased the risk of tumours in the temporal, temporoparietal and occipital areas, with OR 2.42, 95% CI 0.97-6.05 (i.e. the anatomical areas with highest exposure to microwaves from a mobile phone). PMID- 11916352 TI - Epidemiology and aetiological factors of male breast cancer: a ten years retrospective study in eastern Turkey. AB - The aim of this study was to assess the epidemiological and aetiological factors of male breast carcinoma in eastern Turkey. For this purpose we evaluated breast carcinoma patients admitted to our regional hospital from 1990 to 2000. A total of 196 patients were admitted during that time, 11 of whom were male (5%). The average age at presentation was 60.7 +/- 7.5. Infiltrating ductal carcinoma was the most frequent histopathological type; lobular carcinoma was detected in only one of our cases. Right-sided male breast carcinoma was seen in 7 of 11 cases, left-sided in four cases. We detected gynaecomastia in two patients. Other factors were excessive alcohol consumption for 35 years in one patient, family history in one patient and exposure to electromagnetic fields (EMFs) and light at night in four patients. We demonstrated no risk factor in the other three cases. Of the patients in our study, the youngest was 45 years old--the patient with post-pubertal gynaecomastia. The overall rate of male breast carcinoma seen among people who had worked for the Turkish Institution of Electricity in eastern Turkey was 0.3%. In our study we demonstrated a close relation between exposure to EMFs and light at night and male breast carcinoma in eastern Turkey. We also supposed that not only exposure to EMFs but also the duration of the exposure could affect the risk of development of male breast carcinoma. PMID- 11916353 TI - Christmas in science. PMID- 11916354 TI - Breast cancer and PCBs: true or false association? PMID- 11916355 TI - Hypotension after first-dose ACE inhibitor administration in heart failure- should doctors stop worrying? PMID- 11916357 TI - Prior calcium channel blockade and short-term survival following acute myocardial infarction. AB - There is concern over the safety of calcium channel blockers (CCBs) in acute coronary disease. We sought to determine if patients taking calcium channel blockers (CCBs) at the time of admission with acute myocardial infarction (AMI) had a higher case-fatality compared with those taking beta-blockers or neither medication. Clinical and drug treatment variables at the time of hospital admission predictive of survival at 28 days were examined in a community-based registry of patients aged under 65 years admitted to hospital for suspected AMI in Perth, Australia, between 1984 and 1993. Among 7766 patients, 1291 (16.6%) were taking a CCB and 1259 (16.2%) a betablocker alone at hospital admission. Patients taking CCBs had a worse clinical profile than those taking a beta blocker alone or neither drug (control group), and a higher unadjusted 28-day mortality (17.6% versus 9.3% and 11.1% respectively, both P < 0.001). There was no significant heterogeneity with respect to mortality between nifedipine, diltiazem, or verapamil when used alone, or with a beta-blocker. After adjustment for factors predictive of death at 28 days, patients taking a CCB were found not to have an excess chance of death compared with the control group (odds ratio [OR] 1.06, 95% confidence interval [CI]; 0.87, 1.30), whereas those taking a beta blocker alone had a lower odds of death (OR 0.75, 95% CI; 0.59, 0.94). These results indicate that established calcium channel blockade is not associated with an excess risk of death following AMI once other differences between patients are taken into account, but neither does it have the survival advantage seen with prior beta-blocker therapy. PMID- 11916356 TI - Heart function challenged with beta-receptor agonism or antagonism in a heart failure model. AB - We have shown that chronic treatment with carteolol, a non-selective beta adrenergic receptor antagonist, improved left ventricular (LV) function and survival in an avian model of dilated cardiomyopathy (DCM). The aim of the present study was to compare ex vivo heart function with and without beta-agonist and antagonist challenge. We investigated whether intracoronary infusion of a beta-blocker, carteolol or beta-agonist, isoproterenol decreased contractility. In the DCM group, isoproterenol resulted in a significantly greater increase in heart rate (71% vs. 28% compared to control hearts). To investigate the mechanism for the increase in heart rate, we exposed spontaneously beating neonatal cardiomyocytes to serum immunoglobulin (IgG) isolated from DCM animals. Serum IgG resulted in a significant increase in spontaneous beating rate in neonatal rat cardiomyocytes that was blocked by pre-treatment with a beta-blocker. Carteolol challenge did not significantly change heart rate but did significantly increase LV peak pressure in DCM hearts (62%) while coronary artery flow remained unchanged (2.7+/-0.1 vs 2.7+/-0.5 ml/min/g). These results show that 1) beta receptor stimulation results in a greater tachycardic response in DCM animals, and 2) carteolol challenge improves myocardial contractility in hearts from DCM animals independent of heart rate or changes in coronary artery flow. PMID- 11916358 TI - Effect of diltiazem on cardiac function assessed by echocardiography and neurohumoral factors after reperfused myocardial infarction without congestive heart failure. AB - The purpose of this study was to examine the effect of diltiazem on cardiac function and neurohumoral factors (BNP, epinephrine, norepinephrine) after reperfused myocardial infarction without congestive heart failure (Killip class I). On the first day after myocardial infarction following reperfusion therapy patients were randomly assigned to diltiazem treatment (group 1, n=33) or no treatment (group 2, n=39). We then performed echocardiographic examinations on the patients and measured heart rate, mean blood pressure and neurohormones (BNP, epinephrine and norepinephrine). Follow-up evaluations of echocardiography were performed at 4 and 12 weeks and of neurohormones at 1 and 4 weeks after acute myocardial infarction. The highest peaks of plasma BNP, epinephrine, and norepinephrine levels were observed before treatment and decreased with time in both groups. After 4 weeks the level of plasma BNP in the diltiazem treatment group was lower than in the no treatment group [55+/-3 pg/mL vs 85+/-5 pg/mL (P < 0.05)]. Other neurohormones did not differ between groups. Fractional shortening (FS) and ejection fraction (EF)improved after myocardial infarction in both groups, but significantly more in the diltiazem group (P < 0.05) after 12 weeks of treatment. Changes in BNP correlated significantly with changes in left ventricular end systolic volumes, FS and EF. In this study, diltiazem significantly improved systolic function and reduced the level of plasma BNP after myocardial infarction, which suggest that diltiazem may have a beneficial effect on myocardial infarction without congestive heart failure. PMID- 11916359 TI - A comparative study of the first dose hypotensive effects of captopril and perindopril in patients with heart failure. AB - Angiotensin converting enzyme (ACE) inhibitors reduce morbidity and mortality in patients with heart failure and are a first-line therapy for chronic heart failure. However, the first-dose may be associated with asymptomatic or symptomatic hypotension. In previous small series with different ACE inhibitors, different blood pressure responses have been reported. We defined hypotension as a fall in mean blood pressure > or = 20 mm Hg and an absolute value of systolic blood pressure < or = 90 mm Hg and diastolic blood pressure <60 mm Hg. We studied the evolution of mean, systolic and diastolic blood pressure after initiation of perindopril and captopril treatments in a multicentre, double-blind, randomised, comparative, prospective study. One hundred seventy-six patients, mean age 64.9+/ 12.1 years, 116 men, with symptomatic heart failure, NYHA class II-IV, and a left ventricular ejection fraction <40%, were randomised to receive a single dose of captopril 6.25 mg (n = 85) or perindopril 2 mg, (n = 91). Systolic and diastolic blood pressure were recorded with Dinamap every 15 minutes during a baseline period of 2 hours, every 30 minutes from 2 to 7 hours and at 8 hours after the drug administration. Baseline characteristics of both groups were similar (demography, heart failure aetiology, NYHA class and blood pressure). Throughout the study there were 23 asymptomatic episodes of hypotension in the captopril group and 6 in the perindopril group (p = 0.039). One patient in the captopril group had symptomatic episodes. Mean blood pressure falls were significantly higher in the captopril versus perindopril group at 60 minutes (-4.6 mm Hg vs+0.7 mm Hg; p=0.004), 75 minutes (-4.4 mm Hg vs -1.1 mm Hg; p = 0.042), and 180 minutes (-3.4 mm Hg vs +0.0 mm Hg; p= 0.042). When elderly patients (> or =70 years) were considered the same pattern of response was found. In summary, first dose hypotension is not negligible on initiation of therapy with ACE inhibitors in heart failure patients with low ejection fraction. Perindopril results in significantly less reduction in blood pressure and a lower incidence of symptomatic or asymptomatic hypotensive episodes and allows a safer start of therapy than captopril in heart failure patients. PMID- 11916360 TI - Apoptosis in cardiac disease--what is it--how does it occur. AB - This review will present a summary of a description of apoptotic pathways in the heart, followed by ways to measure it and the experimental and clinical evidence for the role of apoptosis in cardiac disease. An evaluation of the effectiveness of pharmacological and other therapeutic interventions in the prevention of apoptosis in the context of cardiac disease will also be presented. PMID- 11916361 TI - Apoptosis in heart failure: a real problem? AB - Progressive left ventricular (LV) dysfunction is a characteristic feature of the failing heart. The mechanism or mechanisms that drive this progression are not known. In recent years, we advanced a working hypothesis that progressive LV dysfunction in heart failure results, in part, from ongoing loss of cardiomyocytes. More recently evidence emerge based on studies in animals with heart failure and in explanted failed human hearts that ongoing cardiomyocyte death through apoptosis occurs in heart failure, a finding that supports the original hypothesis. While these findings created considerable enthusiasm, some skepticism remains even today as to whether cardiomyocyte apoptosis plays an important role in the progression of heart failure. The evidence garnered over the past few years, when considered in aggregate, does favors apoptosis as a key culprit in the progression of the heart failure. Nonetheless, additional key studies are needed to determine if direct inhibition of apoptosis with specific pharmacologic probes prevents progressive LV dysfunction in heart failure. Only then can one fully appreciate the importance of cardiomyocyte apoptosis in the pathophysiology of heart failure. PMID- 11916362 TI - Neurohormones, cytokines and programmed cell death in heart failure: a new paradigm for the remodeling heart. PMID- 11916363 TI - Apoptosis in cardiovascular remodeling--effect of medication. AB - In the last decade, apoptosis has emerged as a key determinant of target organ damage in cardiovascular diseases. The suggestion that increased cardiomyocyte apoptosis participates in the etiology of heart failure probably contributed to the negative view of the prevalence of apoptosis in the field of cardiovascular diseases. However, we and others have shown that up-regulation of apoptosis in certain cardiovascular cells may contribute to the beneficial action of antihypertensive drugs on target-organ structure. As an explanation for this apparent discrepancy, the same stimulus, e.g. angiotensin II, can induce apoptosis or stimulate cell growth in different cell types (e.g., cardiomyocytes and fibroblasts, respectively). Using the angiotensin pathway as a paradigm, this review proposes an integrative view of cell growth and cell death regulation in cardiovascular cells in order to illustrate how cell-specific responses to the same stimulus may in part explain the patterns of cell population dynamics during the development and treatment of target organ damage in hypertension. PMID- 11916364 TI - Apoptosis--new opportunities for novel therapeutics for heart diseases. AB - Apoptosis as defined by contemporary science describes a form of cell death that involves discrete genetic and molecular programs, de novo protein expression and unique cellular phenotype. Evidence for the existence of apoptosis in the human heart has been reported in various cardiac diseases, including ischemic and non ischemic heart failure, myocardial infarction and arrhythmias. Among the most potent stimuli that elicit cardiomyocyte apoptosis are: oxygen radicals (including NO), cytokines, (e.g., TNFalpha, FAS) neurohormonal factors (angiotension II), cardiotoxic drugs (e.g., doxorubicin) and mechanical, stretch situations. Several complex signal transduction pathways have been implicated in execution of cardiomyocyte apoptosis. Most prominent are: 1) Tyrosine kinase receptors (TRK) induced signaling involving stress or mitogen activated protein kinases (SAPK/MARK) and sphingolipids metabolites (ceramide); 2) G-protein coupled receptor (GPCR) signaling (Galphai, Galphaq) and 3) NF(K) B activation. Apoptosis of cardiac myocytes may contribute to progressive pump-failure, arrhythmias and cardiac remodeling. The recognition of diverse molecular targets associated with cardiomyocyte apoptosis provide new opportunities for pharmacologic manipulation, that may lead to discovery and development of therapeutic strategies for treatment of heart failure, arrhythmias and myocardial infarction. PMID- 11916365 TI - Apoptosis in cardiac disease: from basics to clinics--an editorial commentary. PMID- 11916366 TI - The use of simvastatin in analbuminaemia. AB - Congenital analbuminaemia, a rare disorder associated with defective albumin synthesis, is characterised by hyperlipidaemia. Administration of 3-hydroxy-3 methylglutaryl coenzyme A reductase (HMGRI) to analbuminaemic rats have demonstrated no significant effect on plasma lipids, however no published information regarding HMGRI treatment could be found in human subjects. The efficacy, safety and tolerability of Simvastatin was thus investigated in 2 South African patients with analbuminaemia, a 21 year old Caucasian male (H-B) and a 61 year old black male (A-K). In the case of A-K, the lipid profile responded predictably but H-B responded less that expected from general experience with Simvastatin. Both subjects, however, experienced a three- to five-fold increase in creatine kinase. The use of HMGRI's should thus be used cautiously in these patients and it may be advisable to reserve treatment for secondary prevention. PMID- 11916367 TI - Optimizing the prescription of statins after an acute coronary syndrome: the influence of coronary angioplasty and total cholesterol levels. PMID- 11916369 TI - Urine toxicology samples in cocaine treatment trials: how many need to be tested? AB - How frequently should urine samples be collected and analyzed to accurately measure drug use in clinical trials of cocaine abuse treatments? Previous research suggests that analyzing one of three weekly urine toxicology samples in an opiate-related trial may be sufficient. To empirically address this question in the field of cocaine research, we examined the weekly variation in the cocaine metabolite benzoylecgonine (BE) concentration between pairs of weekly urine samples from a clinical trial of a treatment for cocaine dependence. Twice weekly urine samples from 71 subjects collected over eleven weeks were assayed for quantitative BE levels. Agreement between pairs of samples was estimated for both quantitative and qualitative measures of BE. Results indicated substantial intra week variation with correlations never exceeding .50 and approximately 20% disagreement among samples using cutoff values in place of quantitative levels. Both samples, however, supported similar conclusions about group-level behavior. PMID- 11916370 TI - Smokeless tobacco use among addiction patients: a brief report. AB - INTRODUCTION: The prevalence and treatment outcome of smokeless tobacco (ST) use among hospitalized addiction patients is unclear. METHODS: Using initially a retrospective and then a prospective study design, data were collected about patients who described "other tobacco use." The data examined included demographics, medical disorders, psychiatric conditions, nicotine treatment and outcome. RESULTS: The data from the two samples were consistent. All subjects were Caucasian males, mean age 38.5 years, with daily ST use for at least one year. Most had alcohol dependence. The prevalence rate was 3% retrospectively and 7% prospectively. Interventions included nicotine gum and patches. All subjects abstained from ST use during hospitalization but follow-up data were limited. CONCLUSION: These observations are consistent with reports of ST use being more common in males in the general population and its association with alcohol dependence. Further research is needed into effective long-term treatment for ST use among addiction or psychiatric patients. PMID- 11916368 TI - Bupropion treatment for cocaine abuse and adult attention-deficit/hyperactivity disorder. AB - There are few published studies assessing the efficacy of pharmacologic treatments for attention-deficit hyperactivity disorder (ADHD) among substance abusers seeking treatment. Eleven patients who met DSM-IV diagnostic criteria for cocaine dependence and adult ADHD were entered into a 12-week single-blind trial of divided daily doses of bupropion (BPR). All patients received weekly individual standardized relapse prevention therapy. Treatment compliance and retention were good. Patients reported significant reductions in attention difficulties, hyperactivity and impulsivity. Self-reported cocaine use, cocaine craving, and cocaine positive toxicologies, also decreased significantly. In a previously published trial, 12 patients who met similar diagnostic criteria for adult ADHD and cocaine dependence were entered into a 12-week trial of divided daily doses of sustained-release methylphenidate (MPH). Improvements observed on BPR were similar to, and did not differ from those previously observed with MPH. These preliminary data suggest that BPR may be as effective as sustained-release MPH, when combined with relapse prevention therapy, for cocaine abusers with adult ADHD. However, a future study directly comparing BPR to MPH in a double blind placebo-controlled trial is needed. PMID- 11916371 TI - To smoke or not to smoke: impact on disability, quality of life, and illicit drug use in baseline polydrug users. AB - Does tobacco use in polydrug users relate to future daily functioning (SF-36) and disability? To answer this question, community-living polydrug users (n = 254) were interviewed at baseline, 1-year and 2-year follow-up. Measures included: smoking status and self-reported disability at each assessment, and SF-36 measures collected at the final assessment. Urine samples permitted validation of reported drug use status. Results revealed that baseline disability rates were high but fell nearly 50% over two years. Disabilities named were similar to those reported in the general population. Change in smoking status was associated with decreased disability and improvements in general health and vitality. Respondents reporting disability reported lower daily functioning (SF-36). Stable everyday smoking was strongly associated with increased probability of positive urine tests for illicit drug use. Illicit drug use did not affect SF-36 ratings. Findings suggest that tobacco use by polydrug users is related to disability rates, to illicit drug use and to variations in daily functioning. PMID- 11916372 TI - Divalproex sodium (Depakote) for alcohol withdrawal and relapse prevention. AB - This pilot study evaluates the safety and efficacy of divalproex sodium (Depakote) for alcohol withdrawal and relapse prevention. Sixteen patients in moderate alcohol withdrawal were randomized to receive a standard benzodiazepine detoxification, depakote detoxification, or depakote detox plus maintenance. Symptom reduction occurred more rapidly and consistently in the depakote treated patients than in the benzodiazepine control group, and at six-week follow up a greater percentage of patients in the depakote maintenance group were completely abstinent than either detox-only group. There were no significant differences in sociodemographic or drinking data amongst the three cohort samples at baseline. Our findings suggest that the anticonvulsant divalproex sodium (Depakote) may be a safe and efficacious alternative to benzodiazepines for the treatment of alcohol withdrawal. It may be an advantageous alternative for outpatient detoxification, as it has no abuse potential, pharmacologic synergy with alcohol, or substantial cognitive or psychomotor side effects. PMID- 11916373 TI - Hospital Use of Ethanol Survey (HUES): preliminary results. AB - Little information exists about alcohol use within health facilities. We sought to determine alcohol use and control in acute-care hospitals by mailing a questionnaire to a convenience sample of Pharmacy Directors of 24 hospitals in two regions. Of 23 responders, in-patient alcohol was dispensed by 21 (91%) within the last 5 years. Of these 21, both beverage and intravenous alcohol were dispensed by 13 (62%), only beverage alcohol by seven (33%), and only intravenous alcohol by one (5%). No institutional policies regarding alcohol dispensing existed in 16 (70%) hospitals. Alcohol was frequently used as a patient courtesy (14/20, 70%), and to prevent withdrawal (7/20, 35%). All pharmacies procured intravenous alcohol in a formal process, but 60% (12/20) obtained beverage alcohol informally. Alcohol is widely dispensed with few guidelines in this sample of acute-care hospitals. Additional research on therapeutic efficacy, consequences, and institutional oversight of alcohol in hospitals is needed. PMID- 11916374 TI - Improved psychological status in a modified therapeutic community for homeless MICA men. AB - An adaptation of the drug-free therapeutic community (TC) model to homeless men with comorbid mental illness and chemical addiction (MICA) was evaluated with respect to change in psychological status over the course of a six-month residential treatment. Psychological status was assessed by: the Symptom Checklist-90-R (SCL90-R), Beck Depression Inventory (BDI), Shortened Manifest Anxiety Scale (SMAS), and Tennessee Self-Concept Scale (TSCS). A total of 52 out of an original study cohort of 124 residents were followed in longitudinal analyses to treatment midpoint, with a subset of 34 assessed through treatment completion. Significant, widespread psychological improvements were found during both the first and second half of treatment; it would appear that distress reduction was ongoing throughout treatment, with intrapersonal preceding interpersonal relief. The premise of applying a socially-based treatment to this population is discussed in light of these findings. PMID- 11916375 TI - Utilization of preventive health services by HIV-seronegative injection drug users. AB - Little is known about the use of preventive health services by HIV-seronegative IDUs. This study examines screening and preventive health services use by two populations of HIV-seronegative IDUs recruited from a needle exchange program and methadone program. We found high rates of HIV and tuberculin skin testing in the past year as well as tetanus vaccination among both groups of IDUs. Hepatitis B and C testing was significantly higher in the methadone group than the needle exchange group. Syphilis testing and pneumovax administration was disappointingly low in both groups. Given the burden of disease among IDUs, our findings suggest the need for a distinct set of testing and vaccination guidelines for the HIV seronegative IDU population. PMID- 11916376 TI - The genome of bacteriophage phiKZ of Pseudomonas aeruginosa. AB - Bacteriophage phiKZ is a giant virus that efficiently infects Pseudomonas aeruginosa strains pathogenic to human and, therefore, it is attractive for phage therapy. We present here the complete phiKZ genome sequence and a preliminary analysis of its genome structure. The 280,334 bp genome is a linear, circularly permutated and terminally redundant, A+T-rich double-stranded DNA molecule. The phiKZ DNA has no detectable sequence homology to other viruses and microorganisms, and it does not contain NotI, PstI, SacI, SmaI, XhoI, and XmaIII endonuclease restriction sites. The genome has 306 open reading frames (ORFs) varying in size from 50 to 2237 amino acid residues. According to the orientation of transcription, ORFs are apparently organized into clusters and most have a clockwise direction. The phiKZ genome also encodes six tRNAs specific for Met (AUG), Asn (AAC), Asp (GAC), Leu (TTA), Thr (ACA), and Pro (CCA). A putative promoter sequence containing a TATATTAC block was identified. Most potential stem loop transcription terminators contain the tetranucleotide UUCG loops. Some genes may be assigned as phage-encoded RNA polymerase subunits. Only 59 phiKZ gene products exhibit similarity to proteins of known function from a diversity of organisms. Most of these conserved gene products, such as dihydrofolate reductase, ribonucleoside diphosphate reductase, thymidylate synthase, thymidylate kinase, and deoxycytidine triphosphate deaminase are involved in nucleotide metabolism. However, no virus-encoded DNA polymerase, DNA replication associated proteins, or single-stranded DNA-binding protein were found based on amino acid homology, and they may therefore be strongly divergent from known homologous proteins. Fifteen phiKZ gene products show homology to proteins of pathogenic organisms, including Mycobacterium tuberculosis, Haemophilus influenzae, Listeria sp., Rickettsia prowazakeri, and Vibrio cholerae that must be considered before using this phage as a therapeutic agent. The phiKZ coat contains at least 40 polypeptides, and several proteins are cleaved during virus assembly in a way similar to phage T4. Eleven phiKZ-encoded polypeptides are related to proteins of other bacteriphages that infect a variety of hosts. Among these are four gene products that contain a putative intron-encoded endonuclease harboring the H-N-H motif common to many double-stranded DNA phages. These observations provide evidence that phages infecting diverse hosts have had access to a common genetic pool. However, limited homology on the DNA and protein levels indicates that bacteriophage phiKZ represents an evolutionary distinctive branch of the Myoviridae family. PMID- 11916377 TI - Pre-steady-state and stopped-flow fluorescence analysis of Escherichia coli ribonuclease III: insights into mechanism and conformational changes associated with binding and catalysis. AB - To better understand substrate recognition and catalysis by RNase III, we examined steady-state and pre-steady-state reaction kinetics, and changes in intrinsic enzyme fluorescence. The multiple turnover cleavage of a model RNA substrate shows a pre-steady-state burst of product formation followed by a slower phase, indicating that the steady-state reaction rate is not limited by substrate cleavage. RNase III catalyzed hydrolysis is slower at low pH, permitting the use of pre-steady-state kinetics to measure the dissociation constant for formation of the enzyme-substrate complex (K(d)=5.4(+/-0.6) nM), and the rate constant for phosphodiester bond cleavage (k(c)=1.160(+/-0.001) min(-1), pH 5.4). Isotope incorporation analysis shows that a single solvent oxygen atom is incorporated into the 5' phosphate of the RNA product, which demonstrates that the cleavage step is irreversible. Analysis of the pH dependence of the single turnover rate constant, k(c), fits best to a model for two or more titratable groups with pK(a) of ca 5.6, suggesting a role for conserved acidic residues in catalysis. Additionally, we find that k(c) is dependent on the pK(a) value of the hydrated divalent metal ion included in the reaction, providing evidence for participation of a metal ion hydroxide in catalysis, potentially in developing the nucleophile for the hydrolysis reaction. In order to assess whether conformational changes also contribute to the enzyme mechanism, we monitored intrinsic tryptophan fluorescence. During a single round of binding and cleavage by the enzyme we detect a biphasic change in fluorescence. The rate of the initial increase in fluorescence was dependent on substrate concentration yielding a second-order rate constant of 1.0(+/-0.1)x10(8) M(-1) s(-1), while the rate constant of the second phase was concentration independent (6.4(+/-0.8) s( 1); pH 7.3). These data, together with the unique dependence of each phase on divalent metal ion identity and pH, support the hypothesis that the two fluorescence transitions, which we attribute to conformational changes, correlate with substrate binding and catalysis. PMID- 11916378 TI - Classification and evolution of P-loop GTPases and related ATPases. AB - Sequences and available structures were compared for all the widely distributed representatives of the P-loop GTPases and GTPase-related proteins with the aim of constructing an evolutionary classification for this superclass of proteins and reconstructing the principal events in their evolution. The GTPase superclass can be divided into two large classes, each of which has a unique set of sequence and structural signatures (synapomorphies). The first class, designated TRAFAC (after translation factors) includes enzymes involved in translation (initiation, elongation, and release factors), signal transduction (in particular, the extended Ras-like family), cell motility, and intracellular transport. The second class, designated SIMIBI (after signal recognition particle, MinD, and BioD), consists of signal recognition particle (SRP) GTPases, the assemblage of MinD like ATPases, which are involved in protein localization, chromosome partitioning, and membrane transport, and a group of metabolic enzymes with kinase or related phosphate transferase activity. These two classes together contain over 20 distinct families that are further subdivided into 57 subfamilies (ancient lineages) on the basis of conserved sequence motifs, shared structural features, and domain architectures. Ten subfamilies show a universal phyletic distribution compatible with presence in the last universal common ancestor of the extant life forms (LUCA). These include four translation factors, two OBG like GTPases, the YawG/YlqF-like GTPases (these two subfamilies also consist of predicted translation factors), the two signal-recognition-associated GTPases, and the MRP subfamily of MinD-like ATPases. The distribution of nucleotide specificity among the proteins of the GTPase superclass indicates that the common ancestor of the entire superclass was a GTPase and that a secondary switch to ATPase activity has occurred on several independent occasions during evolution. The functions of most GTPases that are traceable to LUCA are associated with translation. However, in contrast to other superclasses of P-loop NTPases (RecA F1/F0, AAA+, helicases, ABC), GTPases do not participate in NTP-dependent nucleic acid unwinding and reorganizing activities. Hence, we hypothesize that the ancestral GTPase was an enzyme with a generic regulatory role in translation, with subsequent diversification resulting in acquisition of diverse functions in transport, protein trafficking, and signaling. In addition to the classification of previously known families of GTPases and related ATPases, we introduce several previously undetected families and describe new functional predictions. PMID- 11916379 TI - The intracellular antibody capture technology (IACT): towards a consensus sequence for intracellular antibodies. AB - We describe the application of an intracellular antibody capture technology (IACT) as a generic in vivo selection procedure for isolating intracellular antibodies or ICAbs. IACT was applied to the de novo selection of functional ICAbs against the microtubule-associated protein TAU, found in neurofibrillary lesions of Alzheimer's disease brains. A panel of 17 different ICAbs was created which bind TAU inside cells and the epitopes recognized by the selected ICAbs have been determined by an in vivo epitope mapping procedure. Finally, sequence analysis showed that the IACT-derived ICAbs are characterized by a common signature of conserved amino acid residues, suggesting that the IACT naturally selects a sort of "captured consensus sequence" for intracellular antibodies. The development of IACT, together with the possibility of scaling up in a high throughput and automated format, makes IACT a new enabling tool for target validation in functional genomics and global proteomics. PMID- 11916381 TI - The crystallographic structure of brome mosaic virus. AB - The structure of brome mosaic virus (BMV), the type member of the bromoviridae family, has been determined from a single rhombohedral crystal by X-ray diffraction, and refined to an R value of 0.237 for data in the range 3.4-40.0 A. The structure, which represents the native, compact form at pH 5.2 in the presence of 0.1 M Mg(2+), was solved by molecular replacement using the model of cowpea chlorotic mottle virus (CCMV), which BMV closely resembles. The BMV model contains amino acid residues 41-189 for the pentameric capsid A subunits, and residues 25-189 and 1-189 for the B and C subunits, respectively, which compose the hexameric capsomeres. In the model there are two Mg ions and one molecule of polyethylene glycol (PEG). The first 25 amino acid residues of the C subunit are modeled as polyalanine. The coat protein has the canonical "jellyroll" beta barrel topology with extended amino-terminal polypeptides as seen in other icosahedral plant viruses. Mass spectrometry shows that in native BMV virions, a significant fraction of the amino-terminal peptides are apparently cleaved. No recognizable nucleic acid residue is visible in the electron density maps except at low resolution where it appears to exhibit a layered arrangement in the virion interior. It is juxtaposed closely with the interior surface of the capsid but does not interpenetrate. The protein subunits forming hexameric capsomeres, and particularly dimers, appear to interact extensively, but the subunits otherwise contact one another sparsely about the 5-fold and quasi 3-fold axes. Thus, the virion appears to be an assembly of loosely associated hexameric capsomeres, which may be the basis for the swelling and dissociation that occurs at neutral pH and elevated salt concentration. A Mg ion is observed to lie exactly on the quasi-3-fold axis and is closely coordinated by side-chains of three quasi symmetry-related residues glutamates 84, with possible participation of side chains from threonines 145, and asparagines 148. A presumptive Mg(2+) is also present on the 5-fold axis where there is a concentration of negatively charged side-chains, but the precise coordination is unclear. In both cases these cations appear to be essential for maintenance of virion stability. Density that is contiguous with the viral interior is present on the 3-fold axis at the center of the hexameric capsomere, where there is a pore of about 6 A diameter. The density cannot be attributed to cations and it was modeled as a PEG molecule. PMID- 11916380 TI - Intracellular antibody capture technology: application to selection of intracellular antibodies recognising the BCR-ABL oncogenic protein. AB - The expression of antibodies inside cells to ablate protein function has the potential for disease therapy and for target validation in functional genomics. However, due to inefficient expression or folding, only a few antibodies or antibody fragments, usually as single-chain Fv antibody fragments (scFv), bind their antigens in an intracellular environment. We have established a genetic selection technology (intracellular antibody capture, IAC) to facilitate the isolation of functional intracellular scFv from a diverse repertoire. This approach comprises an in vitro library screen with scFv-expressing bacteriophage, employing bacterially expressed antigen, followed by a yeast in vivo antibody antigen interaction screen of the sub-library of in vitro scFv antigen-binders. Accordingly, we have isolated panels of scFv that bind intracellularly to the BCR or the ABL parts of the BCR-ABL oncogenic protein. Sequence analysis of the intracellular antibody scFv panels revealed a sequence conservation indicating an intracellular antibody consensus for both VH and VL, which could form the basis for the de novo synthesis of intracellular antibody libraries to be used with intracellular antibody-capture technology. PMID- 11916382 TI - Structure of an extended-spectrum class A beta-lactamase from Proteus vulgaris K1. AB - The structure of a chromosomal extended-spectrum beta-lactamase (ESBL) having the ability to hydrolyze cephalosporins including cefuroxime and ceftazidime has been determined by X-ray crystallography to 1.75 A resolution. The species-specific class A beta-lactamase from Proteus vulgaris K1 was crystallized at pH 6.25 and its structure solved by molecular replacement. Refinement of the model resulted in crystallographic R and R(free) of 16.9 % and 19.3 %, respectively. The folding of the K1 enzyme is broadly similar to that of non-ESBL TEM-type beta-lactamases (2 A rmsd for C(alpha)) and differs by only 0.35 A for all atoms of six conserved residues in the catalytic site. Other residues promoting extended-spectrum activity in K1 include the side-chains of atypical residues Ser237 and Lys276. These side-chains are linked by two water molecules, one of which lies in the position normally filled by the guanidinium group of Arg244, present in most non ESBL enzymes but absent from K1. The ammonium group of Lys276, ca 3.5 A from the virtual Arg244 guanidinium position, may interact with polar R2 substitutents on the dihydrothiazene ring of cephalosporins. PMID- 11916383 TI - Crystallographic and functional studies of a modified form of eosinophil-derived neurotoxin (EDN) with novel biological activities. AB - The crystal structure of a post-translationally modified form of eosinophil derived neurotoxin (EDN) with four extra residues on its N terminus ((-4)EDN) has been solved and refined at atomic resolution (1 A). Two of the extra residues can be placed unambiguously, while the density corresponding to two others is poor. The modified N terminus appears to influence the position of the catalytically important His129, possibly explaining the diminished catalytic activity of this variant. However, (-4)EDN has been shown to be cytotoxic to a Kaposi's sarcoma tumor cell line and other endothelial cell lines. Analysis of the structure and function suggests that the reason for cytotoxicity is most likely due to cellular recognition by the N-terminal extension, since the intrinsic activity of the enzyme is not sufficient for cytotoxicity and the N-terminal extension does not affect the conformation of EDN. PMID- 11916384 TI - Solution structure and dynamics of Crh, the Bacillus subtilis catabolite repression HPr. AB - The solution structure and dynamics of the Bacillus subtilis HPr-like protein, Crh, have been investigated using NMR spectroscopy. Crh exhibits high sequence identity (45 %) to the histidine-containing protein (HPr), a phospho-carrier protein of the phosphoenolpyruvate (PEP):carbohydrate phosphotransferase system, but contains no catalytic His15, the site of PEP-dependent phosphorylation in HPr. Crh also forms a mixture of monomers and dimers in solution whereas HPr is known to be monomeric. Complete backbone and side-chain assignments were obtained for the monomeric form, and 60 % of the dimer backbone resonances; allowing the identification of the Crh dimer interface from chemical-shift mapping. The conformation of Crh was determined to a precision of 0.46(+/-0.06) A for the backbone atoms, and 1.01(+/-0.08) A for the heavy atoms. The monomer structure is similar to that of known HPr 2.67(+/-0.22) A (C(alpha) rmsd), but has a few notable differences, including a change in the orientation of one of the helices (B), and a two-residue shift in beta-sheet pairing of the N-terminal strand with the beta4 strand. This shift results in a shortening of the surface loop present in HPr and consequently provides a flatter surface in the region of dimerisation contact, which may be related to the different oligomeric nature of these two proteins. A binding site of phospho-serine(P-Ser)-Crh with catabolite control protein A (CcpA) is proposed on the basis of highly conserved surface side-chains between Crh and HPr. This binding site is consistent with the model of a dimer dimer interaction between P-Ser-Crh and CcpA. (15)N relaxation measured in the monomeric form also identified differential local mobility in the helix B which is located in the vicinity of this site. PMID- 11916385 TI - Solution structure of protein SRP19 of Archaeoglobus fulgidus signal recognition particle. AB - Protein SRP19 is an essential RNA-binding component of the signal recognition particle (SRP) in Archaea and Eucarya. A three-dimensional solution structure of the 104 residue SRP19 from the hyperthermophilic archaeon Archaeoglobus fulgidus, designated as Af19, was determined by NMR spectroscopy. Af19 contains three beta strands, two alpha-helical regions, arranged in a betaalphabetabetaalpha topology, a 3(10) helix, and a disordered C-terminal tail. This fold is similar to the betaalphabetabetaalphabeta RNP motif present in numerous other RNA-binding proteins, which engage their cognate RNAs using conserved sequence motifs present within beta-strands 1 and 3. Mutagenesis studies of human SRP19, however, reveal the major contact sites with SRP RNA reside within loops 1, 3, and 4. These contacts were verified by the crystal structure of human SRP19 complexed to SRP RNA helix 6 reported subsequent to the submission of the manuscript. The crystal structure also reveals that, unlike canonical RNP motifs, SRP19 does not engage specific RNA bases through conserved sequence motifs present within beta-strands 1 and 3. Instead, SRP19 uses residues both within and flanking beta-strand 1 to stabilize the complex through direct and indirect contacts to the phosphate backbone of the tetraloop, leaving the bases of the tetraloop exposed. This, coupled with the fact that SRP19 appears relatively rigid and undergoes only minor changes in structure upon RNA binding, may underlie the molecular basis by which SRP19 functions to initiate SRP assembly. PMID- 11916387 TI - Controlling the work function of indium tin oxide: differentiating dipolar from local surface effects. AB - Indium tin oxide (ITO) reacts with tetra(tert-butoxy)tin to give surface bound alkoxytin species. Ligand exchange occurs in these surface bound species by reaction with substituted phenols. The speciation of surface phenoxides was measured in ultrahigh vacuum by X-ray photoelectron spectroscopy, as was the work function for the surface modified ITO. It is shown that the molecular dipole moment of the phenol correlates strongly with the measured ITO work function change; no such correlation exists between the acidity of the phenol and this measured change in work function. Results are consistent therefore with an electrostatic model and not with one involving electronegativities of the ligated phenoxide oxygens. PMID- 11916386 TI - Structure of POIA1, a homologous protein to the propeptide of subtilisin: implication for protein foldability and the function as an intramolecular chaperone. AB - Solution structure of POIA1 (Pleurotus ostreatus proteinase A inhibitor 1), which functions as an intramolecular chaperone and as an inhibitor to subtilisin, was determined. By making use of the fact that POIA1 is the only structured protein that shows homology to the propeptide of subtilisin, which is unstructured by itself, foldability of this protein was elucidated. It became clear that the evolutionarily conserved residues play two important roles, one for the maintenance of its own structure, and the other for the interaction with subtilisin. Structural softness and mutational tolerance contained in the POIA1 structure makes it an ideal material for designing a foldable protein. PMID- 11916388 TI - Al-free Sn-Beta zeolite as a catalyst for the selective reduction of carbonyl compounds (Meerwein-Ponndorf-Verley reaction). AB - The catalytic activity of Sn-Beta zeolite in the Meerwein-Ponndorf-Verley (MPV) reduction of carbonyl compounds with secondary alcohols was performed with quantitative yields to the corresponding alcohol. This heterogeneous catalyst exhibits activities and selectivities not observed before with other Me-zeolites. PMID- 11916389 TI - Direct evidence for base-mediated decomposition of alkyl hydroperoxides (ROOH) in the gas phase. AB - The reaction of F(-) with CH(3)OOH has been studied in the gas phase using a tandem flowing afterglow-selected ion flow tube apparatus. The reaction is rapid (k = 1.23 x 10(-9) cm(3) s(-1), 49% efficiency), and formation of HO(-) + CH(2)O + HF is the major reaction channel observed (85%). Isotopic labeling, reactions of F(-) with larger alkyl hydroperoxides, and computational studies demonstrate that the major product ion, HO(-), is formed via a concerted elimination mechanism that appears to be general to all alkyl hydroperoxides possessing an alpha-hydrogen. This mechanism represents a base-mediated decomposition of alkyl hydroperoxides in the gas phase that may have important implications for solution and biochemical reactions. The reverse reaction, CH(3)OO(-) + HF is also efficient (k = 2.43 x 10(-9) cm(3) s(-1)). The major product ensemble HO(-) + CH(2)O + HF (81%) is identical to that of the forward reaction, and represents a novel neutral-catalyzed decomposition of the anion. PMID- 11916390 TI - Unexpected role of O-2 "protecting" groups of glycosyl donors in mediating regioselective glycosidation. AB - Glycosidation of several vicinal diols reveals that exquisite regioselectivity can be achieved by using 2-O-benzoyl n-pentenyl glycoside donors and/or their cyclic 1,2-ortho ester counterparts. The regioselective preferences for both are the same, although ratios and yields may differ. In stark contrast, glycosidation of the diols with the corresponding 2-O-benzylated donors gives poor, if any, regioselectivity. PMID- 11916391 TI - Triple-quantum magic angle spinning (27)Al NMR of aluminum hydroxides. AB - We show that (27)Al triple-quantum magic angle spinning (3Q-MAS) experiments alleviate the second-order quadrupolar broadening to reveal the structure building units of nonequivalent aluminum octahedra in the most extensively studied aluminum hydroxides, namely, gibbsite, bayerite, and boehmite. Further, aided by ab initio calculations of the electric field gradient tensors, the 3Q MAS/MAS results are shown to lead to the assignment of (27)Al isotropic resonances to the aluminum positions in their X-ray-determined structures. The present work paves the way for future studies on various structurally transformed materials derived from these basic aluminum hydroxides. PMID- 11916392 TI - Intermediacy of an N-heterocyclic carbene complex in the catalytic C-H activation of a substituted benzimidazole. AB - An N-heterocyclic carbene complex was found to be the active catalyst in the Rh(I)-catalyzed intramolecular coupling of an alkenyl group to a C-H bond of a substituted benzimidazole. Kinetic studies demonstrated that the catalytic cyclization is zero-order in substrate and first-order in catalyst. Furthermore, DFT calculations with a model system suggest that the rate-limiting step involves insertion of the alkenyl double bond into the rhodium-carbene bond. PMID- 11916393 TI - (13)C[(13)C] 2D NMR: a novel strategy for the study of paramagnetic proteins with slow electronic relaxation rates. AB - Oxidized human [2Fe-2S] ferredoxin has a notably slow electronic relaxation rate, which precludes the observation of signals from nuclei near the iron-sulfur cluster by conventional 2D or 3D methods that utilize proton detection. We have demonstrated the utility of (13)C[(13)C]CT-COSY in identifying connectivity information from fast relaxing carbon nuclei near the paramagnetic center, including those from residues that ligate the metal center. PMID- 11916394 TI - "True" iron(V) and iron(VI) porphyrins: a first theoretical exploration. AB - We present here a first theoretical characterization of iron(V) (S = (3)/(2)) and iron(VI) (S = 0) porphyrin intermediates. The Fe(V) calculations exhibit exceptionally narrow convergence radii and we believe that for this reason they have long eluded researchers working on high-valent iron intermediates. The Fe(V) N(nitrido) bond distance in the DFT(PW91/TZP) optimized geometry of Fe(V)(P)(N) is 1.722 A, comparable to and slightly longer than the Fe(IV)-O bond distance of 1.684 A in Fe(IV)(P)(O) and the Fe(IV)-N(imido) bond distance of 1.698 A in Fe(IV)(P)(NH). In contrast, the Fe(VI)-N(nitrido) bond distances in [Fe(VI)(P)(N)](+) (S = 0) and Fe(VI)(P)(N)(F) (S = 0) are dramatically shorter, 1.508 and 1.533 A, respectively, consistent with the formal triple bond character of the Fe(VI)-N(nitrido) bond. The nitrido ligand appears to be uniquely capable of stabilizing a "true" Fe(V) center, in the sense defined in the paper. All three unpaired electrons in Fe(V)(P)(N) are completely localized on the Fe(V) N(nitrido) axis, with the Fe and N gross atomic spin populations being 1.579 and 1.550, respectively. In contrast, an axial ligand set consisting of an oxide and a fluoride do not stabilize an Fe(V) ground state but favor an electronic structure best described as an Fe(IV)-oxo porphyrin pi-cation radical. PMID- 11916395 TI - Ab initio direct dynamics study of cyclopropyl radical ring-opening. AB - Quasiclassical direct dynamics simulations, at the CASSCF(3,3)/6-31G(d) level of theory, are used to study the stereochemistry of the electrocyclic ring-opening reaction of the cyclopropyl radical. The trajectories are initiated at the reaction's transition state (TS), with their initial conditions sampled from the TS's 174 degrees C Boltzmann distribution. Intrinsic reaction coordinate calculations predict the overall reaction to have disrotatory stereochemistry. Though this is the preferred initial reaction stereochemistry in the trajectories, 43% of the trajectories follow the conrotatory path. Four unique trajectory types are observed during 200 fs dynamics of the product allyl radical. Intramolecular vibrational energy redistribution and internal rotation are incomplete on this time scale, and a statistical distribution of the allyl isomers is not observed. PMID- 11916396 TI - Engineering an obligate domain-swapped dimer of cyanovirin-N with enhanced anti HIV activity. AB - The anti-HIV cyanobacterial protein cyanovirin-N can undergo domain swapping to form an intertwined dimer. The dimeric form is stable at low pH and millimolar concentrations. By deleting an amino acid from the hinge linker about which domain swapping occurs, we have constructed an obligate domain-swapped dimer of cyanovirin-N that represents a new tetravalent carbohydrate binding protein that is stable over a large range of pH values. This obligate dimer displays enhanced anti-HIV activity relative to the wild-type cyanovirin-N monomer with an observed 3.5-fold decrease in IC(50) (9nM for the dimer vs 32 nM for the monomer) for inhibition of HIV-1 envelope-mediated cell fusion and, when expressed in Escherichia coli, can be rapidly obtained in >98% purity in a single chromatographic step. PMID- 11916397 TI - A trinickel dipyridylamido complex with metal-metal bonding interaction: prelude to polynickel molecular wires and devices? AB - The molecule Ni(3)(dpa)(4)Cl(2) (1) can be oxidized by AgPF(6) to give crystalline Ni(3)(dpa)(4)(PF(6))(3) (2) (dpa is the anion of di(2-pyridyl)amine). This reversible oxidation occurs at a potential of 0.908 V vs Ag/AgCl electrochemically. The X-ray structure of 2 shows that the oxidation causes a major structural change (even though it is reversible), namely, a contraction of the Ni-Ni distances from ca. 2.43 A to 2.284[1] A. In addition, the electronic structure changes so that from four unpaired electrons in 1 there is only one in 2. From these remarkable results, it is inferred that while 1, and all higher homologues with 5, 7, 9,... nickel atoms are poor electronic conductors, the cations obtainable from them may be much better ones. This in turn means that by controlling the oxidation state electrochemically, these molecules may be able to function as nanoscale diodes. PMID- 11916398 TI - Two-dimensional (13)C-(13)C correlation spectroscopy with magic angle spinning and dynamic nuclear polarization. AB - The sensitivity of solid-state NMR experiments can be enhanced with dynamic nuclear polarization (DNP), a technique that transfers the high Boltzmann polarization of unpaired electrons to nuclei. Signal enhancements of up to 23 have been obtained for magic angle spinning (MAS) experiments at 5 T and 85-90 K using a custom-designed high-power gyrotron. The extended stability of MAS/DNP experiments at low temperature is demonstrated with (1)H-driven (13)C spin diffusion experiments on the amino acid proline. These (13)C-(13)C chemical shift correlation spectra are the first two-dimensional MAS/DNP experiments performed at high field (>1.4 T). PMID- 11916399 TI - Formation and structure of a novel enediyne-RNA base covalent adduct. AB - The structure of an unusual covalent adduct formed by thiol-activated neocarzinostatin chromophore (NCS-chrom) and a RNA-DNA hybrid having an overhang of four unpaired residues at the 3'-end of the RNA strand has been elucidated by MS and NMR spectroscopic analyses. Unlike previously characterized adducts formed by NCS-chrom on the sugar residue of the DNA target, this adduct has been found to be on one of the uracil bases in the RNA overhang. Covalent linkage is between C-6 of the post-activated NCS-chrom and C-5 of the uracil. A novel mechanism involving adduction of the NCS-chrom C-6 radical, generated by 2-mercaptoethanol activation, to C-5 of the uracil at the U9 position of the RNA 11-mer, oxidation by dioxygen, reduction by the thiol, and subsequent dehydration is proposed for adduct formation. PMID- 11916400 TI - Single flexible hydrophobic polyelectrolyte molecules adsorbed on solid substrate: transition between a stretched chain, necklace-like conformation and a globule. AB - Single flexible polyelectrolyte molecules of poly(2-vinylpyridine) undergo conformational transition from a stretched wormlike coil to a necklace-like globule, and to a compact globule depending on pH and ionic strength in aqueous solution in good agreement with recent theoretical reports. AFM investigations allow the visualization of details of the chain conformation on mica and the extraction of quantitative statistics of molecular dimensions. PMID- 11916402 TI - Built to order: molecular tinkertoys from the [Re(6)(mu(3)-Se)(8)](2+) clusters. AB - Ligand substitution of [Re(6)(mu(3)-Se)(8)(PEt(3))(5)(CH(3)CN)](SbF(6))(2) (1) with pyridyl-based ligands, 2,4,6-tri-4-pyridyl-1,3,5-triazine (L1) and 5,10,15,20-tetra(4-pyridyl)-21H,23H-porphine (L2), produced respectively the star shaped tricluster (T1) and tetracluster (T2) arrays, wherein three (T1) and four (T2) units of the [Re(6)(mu(3)-Se)(8)](2+) core-containing clusters are interconnected by the corresponding bridging ligands. These novel supramolecular assemblies were characterized by a combination of NMR ((1)H and (31)P) spectroscopy, ESI-MS, and microanalysis. The molecular and solid-state structures of T1 have also been established by single-crystal X-ray diffraction. PMID- 11916401 TI - Aqueous aldol catalysis by a nicotine metabolite. AB - Nornicotine, an endogenous tobacco alkaloid and minor nicotine metabolite, can catalyze aldol reactions at physiological pH. Catalysis appears to be due to a covalent enamine mechanism, an unprecedented reaction with small organic molecule catalysts in aqueous buffer. Kinetic parameters for nornicotine as well as other related alkaloids were measured and demonstrate that both the pyrrolidine and pyridine rings are critical for optimal catalysis. Substrate compatibility of this catalyst and its implications in vivo are discussed. PMID- 11916403 TI - Ring expansion via olefin metathesis. AB - Various macrocycles were prepared in one step by a novel ring-expansion method using olefin metathesis. PMID- 11916404 TI - TpPt(IV)Me(H)(2) forms a sigma-CH(4) complex that is kinetically resistant to methane liberation. AB - Complex 1 undergoes H/D scrambling in methanol without concomitant liberation of either methane or dihydrogen (k(H)/k(D) = 0.76, 55 degrees C). The measured isotope effect was proposed to directly relate to the initial reductive coupling step in reductive elimination reactions. PMID- 11916405 TI - Design of novel antibiotics that bind to the ribosomal acyltransfer site. AB - The structure of neamine bound to the A site of the bacterial ribosomal RNA was used in the design of novel aminoglycosides. The design took into account stereo and electronic contributions to interactions between RNA and aminoglycosides, as well as a random search of 273 000 compounds from the Cambridge structural database and the National Cancer Institute 3-D database that would fit in the ribosomal aminoglycoside-binding pocket. A total of seven compounds were designed and subsequently synthesized, with the expectation that they would bind to the A site RNA. Indeed, all synthetic compounds were found to bind to the target RNA comparably to the parent antibiotic neamine, with dissociation constants in the lower micromolar range. The synthetic compounds were evaluated for antibacterial activity against a set of important pathogenic bacteria. These designer antibiotics showed considerably enhanced antibacterial activities against these pathogens, including organisms that hyperexpressed resistance enzymes to aminoglycosides. Furthermore, analyses of four of the synthetic compounds with two important purified resistance enzymes for aminoglycosides indicated that the compounds were very poor substrates; hence the activity of these synthetic antibiotics does not appear to be compromised by the existing resistance mechanisms, as supported by both in vivo and in vitro experiments. The design principles disclosed herein hold the promise of the generation of a large series of designer antibiotics uncompromised by the existing mechanisms of resistance. PMID- 11916406 TI - Characterization of an RNA active site: interactions between a Diels-Alderase ribozyme and its substrates and products. AB - Ribozymes have recently been shown to catalyze the stereoselective formation of carbon-carbon bonds between small organic molecules. The interactions of these Diels-Alderase ribozymes with their substrates and products have now been elucidated by chemical substitution analysis by using 44 different, systematically varied analogues. RNA-diene interaction is governed by stacking interactions, while hydrogen bonding and metal ion coordination appear to be less important. The diene has to be an anthracene derivative, and substituents at defined positions are permitted, thereby shedding light on the geometry of the binding site. The dienophile must be a five-membered maleimidyl ring with an unsubstituted reactive double bond, and a hydrophobic side chain makes a major contribution to RNA binding. The ribozyme distinguishes between different enantiomers of chiral substrates and accelerates cycloadditions with both enantio and diastereoselectivity. The stereochemistry of the reaction is controlled by RNA-diene interactions. The RNA interacts strongly and stereoselectively with the cycloaddition products, requiring several structural features to be present. Taken together, the results highlight the intricacy of ribozyme active sites which can control chemical reaction pathways based on minute differences in substrate stereochemistry and substitution pattern. PMID- 11916407 TI - Total synthesis and initial structure-function analysis of the potent V-ATPase inhibitors salicylihalamide A and related compounds. AB - Salicylihalamide A is the first member of a growing class of macrocyclic salicylate natural products that induce a variety of interesting phenotypes in cultured mammalian cells. Salicylihalamide A was reported to be a unique and highly differential cytotoxin and a potent inhibitor of the mammalian vacuolar (H(+))-ATPase. The total synthesis of both enantiomers of salicylihalamide A, a revision of the absolute configuration assigned to the natural product, and extensive structure-function studies with synthetic salicylihalamide variants are reported. These studies were possible only due to a highly efficient synthetic strategy that features (1) a remarkably E-selective ring-closing olefin metathesis to construct the 12-membered benzolactone skeleton 29, (2) a mild stereocontrolled elaboration to E-alkenyl isocyanate 41, and (3) addition of carbon, oxygen, and sulfur nucleophiles to isocyanate 41 to obtain salicylihalamide A and congeners. We demonstrate for the first time that salicylihalamide A is a potent inhibitor of fully purified reconstituted V-ATPase from bovine brain, and have identified several similarly potent side chain modified derivatives, including salicylihalamide dimers 43-45. In combination, these studies have laid the foundation for ongoing studies aimed at a comprehensive understanding of salicylihalamide's mode-of-action, of potential relevance to the development of lead compounds for the treatment of osteoporosis and cancer. PMID- 11916408 TI - Substituted 1,8-naphthyridin-2(1H)-ones are superior to thymine in the recognition of adenine in duplex as well as triplex structures. AB - The synthesis and evaluation of a series of novel nucleobases based on substituted 1,8-naphthyridin-2(1H)-ones are reported. The nucleobases were designed to meet the requirements for incorporation into peptide nucleic acids (PNAs) and were evaluated as part of PNA duplex and triplex nucleic acid recognition systems. Of the various nucleobases tested, only the 7-chloro-1,8 naphthyridin-2(1H)-one (7-Cl-bT) nucleobase led to consistently increased affinity in all recognition systems, duplex (Watson-Crick) as well as triplex (Hoogsteen). For multiply modified systems, the increase in thermal stability per modification was dependent on the sequence context, ranging from 2.0 degrees C (in separate positions) to 3.5 degrees C (in adjacent positions) in PNA-DNA duplexes and from 1.2 degrees C (in separate positions) to 3.2 degrees C (in adjacent positions) in PNA-RNA duplexes. Singly mismatched oligonucleotide targets were employed to demonstrate uncompromised sequence discrimination. When part of multiply modified triplex (Hoogsteen) recognition systems, the 7-Cl-bT unit gave rise to increases in the thermal stability ranging from 2.7 to 3.5 degrees C when incorporated into separated and adjacent positions, respectively. Our results furthermore indicate that the duplex stabilization is predominantly enthalpic and therefore most likely not a consequence of single-strand preorganization. Finally, and most surprisingly, we find no direct correlation between the end-stacking efficiency of this type of nucleobase and its helix stabilization when involved in Watson-Crick base pairing within a helix. PMID- 11916409 TI - Synthesis and characterization of oligodeoxynucleotides containing formamidopyrimidine lesions and nonhydrolyzable analogues. AB - Oligodeoxynucleotides containing formamidopyrimidine lesions and C-nucleoside analogues at defined sites were prepared by solid-phase synthesis and in some cases enzymatic ligation. Formamidopyrimidine lesions were introduced as dinucleotides to prevent rearrangement to their pyranose isomers. Oligodeoxynucleotides containing single diastereomers of C-nucleoside analogues of Fapy.dA were introduced by using the respective phosphoramidites. The formamidopyrimidine lesions reduce the T(M) of dodecamers relative to their unmodified nucleotide counterparts when opposite the nucleotide proper base pairing partner. However, duplexes containing Fapy.dG-dA mispairs melt significantly higher than those comprised of dG-dA. All duplexes containing Fapy.dA-dX or its C-nucleoside analogue melt lower than the respective complexes containing dA-dX. Studies of the alkaline lability of oligodeoxynucleotides containing formamidopyrimidine lesions indicate that Fapy.dA is readily identified as an alkali-labile lesion with use of piperidine (1.0 M, 90 degrees C, 20 min), but Fapy.dG is less easily identified in this manner. PMID- 11916410 TI - Identification of a protein-promoting vibration in the reaction catalyzed by horse liver alcohol dehydrogenase. AB - In this article we present computational studies of horse liver alcohol dehydrogenase (HLADH). The computations identify a rate-promoting vibration that is symmetrically coupled to the reaction coordinate. In HLADH a bulky amino acid (Val203) is positioned at the face of the nicotinamide adenine dinucleotide (NAD(+)) cofactor distal to alcohol substrate to restrict the separation of reactants and control the stereochemistry. Molecular dynamics simulations were performed on the dimeric HLADH, including the NAD cofactor, the substrate, and the crystallographic waters, for three different configurations, reactants, products, and transition state. From the spectral density for the substrate-NAD relative motion, and that for the NAD-Val203 relative motion, we find that the two motions are in resonance. By computing the associated spectrum, we find that the reaction coordinate is coupled with the substrate-NAD motion, and from the fact that the coupling vanishes at or near the transition state (demonstrated by the disappearance of strong features in the spectral density), we conclude that the substrate-NAD motion plays the role of a promoting vibration symmetrically coupled to the reaction coordinate. PMID- 11916411 TI - Exploring the hydroxylation-dehydrogenation connection: novel catalytic activity of castor stearoyl-ACP Delta(9) desaturase. AB - The novel product profile obtained by incubating chiral fluorinated substrate analogues with castor stearoyl-ACP Delta(9) desaturase has been rationalized through a series of labeling studies. It was found that the introduction of the Z double bond between C-9 and C-10 of the parent substrate occurs with pro-R enantioselectivity--a result that accounts for the observed stereochemistry of oxidation products derived from (9R)- and (9S)-9-fluorostearoyl-ACP. Oxidation of (9R)-9-fluorostearoyl-ACP occurs via at least two rapidly interchanging substrate conformations in the active site as detected by reaction pathway branching induced by deuteration at C-10 and C-11. Hydroxylation and desaturation of this substrate share the same site of initial oxidative attack. PMID- 11916412 TI - Cation-pi interaction in model alpha-helical peptides. AB - Cation-pi interactions are increasingly recognized as important in chemistry and biology. Here we investigate the cation-pi interaction by determining its effect on the helicity of model peptides using a combination of CD and NMR spectroscopy. The data show that a single Trp/Arg interaction on the surface of a peptide can make a significant net favorable free energy contribution to helix stability if the two residues are positioned with appropriate spacing and orientation. The solvent-exposed Trp-->Arg (i, i + 4) interaction in helices can contribute -0.4 kcal/mol to the helix stability, while no free energy gain is detected if the two residues have the reversed orientation, Arg-->Trp (i, i + 4). The derived free energy is consistent with other experimental results studied in proteins or model peptides on cation-pi interactions. However in the same system the postulated Phe/Arg (i, i + 4) cation-pi interaction provides no net free energy to helix stability. Thus the Trp-->Arg interaction is stronger than Phe-->Arg. The cation pi interactions are not sensitive to the screening effect by adding neutral salt as indicated by salt titration. Our results are in qualitative agreement with theoretical calculations emphasizing that cation-pi interactions can contribute significantly to protein stability with the order Trp > Phe. However, our and other experimental values are significantly smaller than estimates from theoretical calculations. PMID- 11916413 TI - Highly anti-selective asymmetric aldol reactions using chiral zirconium catalysts. Improvement of activities, structure of the novel zirconium complexes, and effect of a small amount of water for the preparation of the catalysts. AB - Catalytic asymmetric aldol reactions of silyl enol ethers with aldehydes (Mukaiyama aldol reactions) have been performed using novel chiral zirconium catalysts. The reactions proceeded in high yields under mild conditions, and anti adducts were obtained in high diastereo- and enantioselectivities. The catalysts were first prepared from zirconium(IV) tert-butoxide (Zr(O(t)Bu)(4)), (R)-3,3' diiodo-1,1'-binaphthalene-2,2'-diol ((R)-3,3'-I(2)BINOL), a primary alcohol, and a small amount of water. It was revealed that the primary alcohol played an important role in completing the catalytic cycle and that a small amount of water was essential for obtaining high selectivities. Moreover, activities of the chiral zirconium catalysts were enhanced by using new ligands, (R)-3,3'-I(2)-6,6' X(2)BINOL (X = Br, I, C(2)F(5)), and it has been shown that even aldol reactions of less reactive substrates proceeded smoothly using the novel zirconium catalysts. Finally, NMR studies of these catalysts were performed, which suggested that the catalyst would form a dimeric structure and that the water affected the catalyst formation. PMID- 11916414 TI - Activation of acyl phosphate monoesters by lanthanide ions: enhanced reactivity of benzoyl methyl phosphate. AB - Acyl phosphate monoesters are intermediates in many biochemical acylation reactions, such as those involving aminoacyl adenylates. Benzoyl methyl phosphate, a typical acyl phosphate monoester, is slowly hydrolyzed in neutral solutions but reacts rapidly with amines. Since biochemical processes of acyl phosphate monoesters involve accelerated reactions with oxygen-centered nucleophiles, we sought catalysts for hydrolysis and methanolysis of benzoyl methyl phosphate to mimic the biochemical outcome. Lanthanide ions are particularly effective catalysts, accelerating reactions much more than comparable levels of magnesium ion. Detailed kinetic analysis of the hydrolysis reactions reveals formation of a 1:1 complex, followed by rapid reaction with a nucleophile. The hydroxide-dependent hydrolysis rate in the europium complex is about 10(5) times that of free substrate with hydroxide. A mechanism that accounts for the data and observed behavior involves bidentate coordination of the metal ion by the acyl phosphate through phosphate and carbonyl oxygens, lowering the energy of the tetrahedral addition intermediate and the associated transition states. The dependence of the metal ion catalyzed process on the concentration of hydroxide ion is consistent with coordinated hydroxide acting as a nucleophile. The reaction of benzoyl methyl phosphate with methanol to form methyl benzoate and methyl phosphate is 30 000 times more rapid in the presence of 0.0001 M lanthanum triflate (in the absence of the metal ion k(obs) = 2.1 x 10(-7) s(-1), at 25 degrees C). Thus, the combination of acyl phosphate esters and lanthanide salts appears to be a promising method for biomimetic acylation of hydroxyl groups. PMID- 11916415 TI - Ligand-modulated stereo- and regioselective tandem addition reactions of rhenium bound naphthalene. AB - A series of complexes of the form TpRe(CO)(L)(eta(2)-naphthalene) (Tp = hydridotris(pyrazolyl)borate) undergoes tandem electrophile/nucleophile addition reactions with a high degree of regiocontrol depending on the auxiliary ligand, L. When L = PMe(3), the reaction of the eta(2)-naphthalene complex with triflic acid followed by a silyl ketene acetal favors the 1,4-addition product, whereas when L = pyridine, N,N-dimethylaminopyridine, N-methylimidazole, or NH(3) the 1,2 addition product is favored. These reactions proceed with excellent stereocontrol: both electrophile (H(+), D(+)) and nucleophile (silyl ketene acetal) add anti to the face of metal coordination, and a single coordination diastereomer can be isolated for each reaction. One-electron oxidation of the Re complex affords the corresponding free dihydronaphthalene in good yield. PMID- 11916416 TI - Formation of cyclodimeric (sp(2)-C(1))-bridged Cp/-oxido ("CpC(1)O"M(IV)X(2)) group 4 metal Ziegler-Natta catalyst systems--how important is the "constrained geometry" effect? AB - Deprotonation of sodium acetylcyclopentadienide (11) was achieved by treatment with LDA in THF to generate the dianion equivalent [Cp-C(=CH(2))-O](2-)(12). Transmetalation with Cl(2)Ti(NMe(2))(2) gave ([Cp-C(=CH(2))-O]Ti(NMe(2))(2))(2) (17); treatment of 12 with Cl(2)Zr(NEt(2))(2)(THF)(2) furnished (([Cp-C(=CH(2)) O]Zr(NEt(2))(2))(2) (18). Cryoscopy in benzene revealed a dimeric structure of 18 in solution. Complex 18 was characterized further by an X-ray crystal structure analysis and by DFT calculations. The two zirconium centers of 18 are connected by means of two symmetry-equivalent eta(5):kappaO[Cp-C(=CH(2))-O] ligands. The ligand backbone shows no specific steric constraints, different from the formally related "constrained geometry" systems such as [Cp-SiMe(2)-NCMe(3)]Zr(NMe(2))(2) (1b). Nevertheless, upon treatment with MAO the CpCO group 4 metal complex system (18) generates an active homogeneous Ziegler-Natta catalyst for effective ethene/1-octene copolymerization, with up to 20% 1-octene having become incorporated in the resulting copolymer at 90 degrees C. PMID- 11916417 TI - Living polymerization of ethylene catalyzed by titanium complexes having fluorine containing phenoxy-imine chelate ligands. AB - Seven titanium complexes bearing fluorine-containing phenoxy-imine chelate ligands, TiCl(2)[eta(2)-1-[C(H)=NR]-2-O-3-(t)Bu-C(6)H(3)](2) [R = 2,3,4,5,6 pentafluorophenyl (1), R = 2,4,6-trifluorophenyl (2), R = 2,6-difluorophenyl (3), R = 2-fluorophenyl (4), R = 3,4,5-trifluorophenyl (5), R = 3,5-difluorophenyl (6), R = 4-fluorophenyl (7)], were synthesized from the lithium salt of the requisite ligand and TiCl(4) in good yields (22%-76%). X-ray analysis revealed that the complexes 1 and 3 adopt a distorted octahedral structure in which the two phenoxy oxygens are situated in the trans-position while the two imine nitrogens and the two chlorine atoms are located cis to one another, the same spatial disposition as that for the corresponding nonfluorinated complex. Although the Ti-O, Ti-N, and Ti-Cl bond distances for complexes 1 and 3 are very similar to those for the nonfluorinated complex, the bond angles between the ligands (e.g., O-Ti-O, N-Ti-N, and Cl-Ti-Cl) and the Ti-N-C-C torsion angles involving the phenyl on the imine nitrogen are different from those for the nonfluorinated complex, as a result of the introduction of fluorine atoms. Complex 1/methylalumoxane (MAO) catalyst system promoted living ethylene polymerization to produce high molecular weight polyethylenes (M(n) > 400 000) with extremely narrow polydispersities (M(w)/M(n) < 1.20). Very high activities (TOF > 20 000 min(-1) atm(-1)) were observed that are comparable to those of Cp(2)ZrCl(2)/MAO at high polymerization temperatures (25, 50 degrees C). Complexes 2-4, which have a fluorine atom adjacent to the imine nitrogen, behaved as living ethylene polymerization catalysts at 50 degrees C, whereas complexes 5 7, possessing no fluorine adjacent to the imine nitrogen, produced polyethylenes having M(w)/M(n) values of ca. 2 with beta-hydrogen transfer as the main termination pathway. These results together with DFT calculations suggested that the presence of a fluorine atom adjacent to the imine nitrogen is a requirement for the high-temperature living polymerization, and the fluorine of the active species for ethylene polymerization interacts with a beta-hydrogen of a polymer chain, resulting in the prevention of beta-hydrogen transfer. This catalyst system was used for the synthesis of a number of unique block copolymers such as polyethylene-b-poly(ethylene-co-propylene) diblock copolymer and polyethylene-b poly(ethylene-co-propylene)-b-syndiotactic polypropylene triblock copolymer from ethylene and propylene. PMID- 11916418 TI - A first transition series pseudotetrahedral oxynitride anion: synthesis and characterization of Ba(2)VO(3)N. AB - Ammonolysis of reactive oxide precursors affords the vanadium(V) phase Ba(2)VO(3)N that is shown by X-ray and neutron powder diffraction and Raman spectroscopy to contain pseudotetrahedral VO(3)N(4-) anions. This is the first example of such species for the first transition series metals. PMID- 11916419 TI - Nearly monodisperse and shape-controlled CdSe nanocrystals via alternative routes: nucleation and growth. AB - The nucleation and growth of colloidal CdSe nanocrystals with a variety of elongated shapes were explored in detail. The critical size nuclei for the system were magic sized nanoclusters, which possessed a sharp and dominated absorption peak at 349 nm. The formation of the unique magic sized nuclei in a broad monomer concentration range was not expected by the classic nucleation theory. We propose that this was a result of the extremely high chemical potential environment, that is, very high monomer concentrations in the solution, required for the growth of those elongated nanocrystals. The shape, size, and size/shape distributions of the resulting nanocrystals were all determined by two related factors, the magic sized nuclei and the concentration of the remaining monomers after the initial nucleation stage. Without any size sorting, nearly monodisperse CdSe quantum structures with different shapes were reproducibly synthesized by using the alternative cadmium precursors, cadmium-phosphonic acid complexes. A reasonably large excess of the cadmium precursor, which is less reactive than the Se precursor, was found beneficial for the system to reach the desired balance between nucleation and growth. The shape evolution and growth kinetics of these elongated nanocrystals were consistent with the diffusion-controlled model proposed previously. The branched nanocrystals had to grow at very high monomer concentrations because the multiple growth centers at the end of each branch must be fed with a very high diffusion flux to keep all branches in the 1D-growth mode. The rice-shaped nanocrystals were found as special products of the 3D growth stage. The growth of the nanocrystals in the 1D-growth stage was proven to be not unidirectional after the length of the nanocrystals reached a certain threshold. Experimental results indicate that coordinating solvents and two ligands with distinguishable coordinating abilities are both not intrinsic requirements for the growth of elongated CdSe nanocrystals. PMID- 11916420 TI - Enhancement of twisting power in the chiral nematic phase by introducing achiral banana-shaped molecules. AB - Achiral banana-shaped molecules with dodecyloxy tail groups, P-12-O-PIMB, N-12-O PIMB, and S-12-O-PIMB, have exhibited unusual smectic phases which possess chiral and helical structures. In this work, we mixed these banana-shaped molecules with the chiral molecule forming a chiral nematic liquid crystal and found an exclusive effect of the achiral dopant that the twisting power of the chiral nematic phase in the mixtures is significantly increased with the increase of the content of achiral banana-shaped molecules. This characteristic effect in the chiral nematic field seems to offer the rational evidence for the twist conformation of such banana-shaped molecules, since the chirality should be included intrinsically within each chain. The asymmetric twist conformation in the ester linkage group connecting the central core with the side wings is likely to be the origin of enhanced twisting power. PMID- 11916421 TI - Naked clusters of 56 tin atoms in the solid state. AB - Isolated, gigantic tin clusters of 56 atoms are discovered in the ternary compound Ba(16)Na(204)Sn(310) (cubic, F(-)43m, Z = 1, a = 25.2041(8) A) made by direct fusion of the elements at 800 degrees C. The cluster, made of four face fused pentagonal dodecahedra, has 36 pentagonal faces and 90 edges, and resembles a concave fullerene "dented" at four places. It is made of three- and four-bonded tin atoms and is "stuffed" with four barium cations, [Ba(4)@Sn(56)](36-). This is the largest main group naked cluster in the solid state besides the fullerenes. Also occurring in the structure are two other isolated clusters of tin, Sn(16-n) (n = 0, 1, 2, 3, or 4) and Sn(8). PMID- 11916422 TI - Intercalators. 1. Nature of stacking interactions between intercalators (ethidium, daunomycin, ellipticine, and 4',6-diaminide-2-phenylindole) and DNA base pairs. Ab initio quantum chemical, density functional theory, and empirical potential study. AB - Properties of isolated intercalators (ethidium (E), daunomycin (D), ellipticine (EL), and 4,6'-diaminide-2-phenylindole (DAPI)) and their stacking interactions with adenine...thymine (AT) and guanine...cytosine (GC) nucleic acid base pairs were investigated by means of a nonempirical correlated ab initio method. All intercalators exhibit large charge delocalization, and none of them (including the DAPI dication) exhibits a site with dominant charge. All intercalators have large polarizability and are good electron acceptors, while base pairs are good electron donors. MP2/6-31G*(0.25) stabilization energies of intercalator...base pair complexes are large (E...AT, 22.4 kcal/mol; D...GC, 17.8 kcal/mol; EL...GC, 18.2 kcal/mol; DAPI...GC, 21.1 kcal/mol) and are well reproduced by modified AMBER potential (van der Waals radii of intercalator atoms are enlarged and their energy depths are increased). Standard AMBER potential underestimates binding, especially for DAPI-containing complexes. Because the DAPI dication is the best electron acceptor (among all intercalators studied), this difference is explained by the importance of the charge-transfer term, which is not included in the AMBER potential. For the neutral EL molecule, the standard AMBER force field provides correct results. The Hartree-Fock and DFT/B3LYP methods, not covering the dispersion energy, fail completely to reveal any energy minimum at the potential energy curve of the E...AT complex, and these methods thus cannot be recommended for a study of intercalation process. On the other hand, an approximate version of the DFT method, which was extended to cover London dispersion energy, yields for all complexes very good stabilization energies that are well comparable with referenced ab initio data. Besides the vertical dependence of the interaction, an energy twist dependence of the interaction energy was also investigated by a reference correlated ab initio method and empirical potentials. It is concluded that, despite the cationic (E +1, D +1, DAPI +2) or polar (EL) character of the intercalators investigated, it is the dispersion energy which predominantly contributes to the stability of intercalator...base pair complexes. Any procedure which does not cover dispersion energy is thus not suitable for studying the process of intercalation. PMID- 11916423 TI - Dynamics of alkane hydroxylation at the non-heme diiron center in methane monooxygenase. AB - Semiclassical molecular dynamics simulations have been combined with quantum chemistry calculations to provide detailed modeling of the methane and ethane hydroxylation reactions catalyzed by the hydroxylase enzymes of the soluble methane monooxygenase system. The experimental distribution of enantiomeric alcohols in the reaction of ethanes made chiral by the use of hydrogen isotopes is quantitatively reproduced and explained. The reaction dynamics involve a mixture of concerted and bound radical trajectories, and we characterize each of these reactive channels in detail. Diffusion of the bound radical intermediate at the active site core determines the global rate constant. The results also provide a qualitative rationale for the lack of ring-opened products derived from certain radical clock substrate probes and for the relative rate constants and kinetic isotope effects exhibited by a variety of substrates. PMID- 11916424 TI - Heme distortions in sperm-whale carbonmonoxy myoglobin: correlations between rotational strengths and heme distortions in MD-generated structures. AB - We have investigated the effects of heme rotational isomerism in sperm-whale carbonmonoxymyoglobin using computational techniques. Several molecular dynamics simulations have been performed for the two rotational isomers A and B, which are related by a 180 degrees rotation around the alpha-gamma axis of the heme, of sperm-whale carbonmonoxy myoglobin in water. Both neutron diffraction and NMR structures were used as starting structures. In the absence of an experimental structure, the structure of isomer B was generated by rotating the heme in the structure of isomer A. Distortions of the heme from planarity were characterized by normal coordinate structural decomposition and by the angle of twist of the pyrrole rings from the heme plane. The heme distortions of the neutron diffraction structure were conserved in the MD trajectories, but in the NMR-based trajectories, where the heme distortions are less well defined, they differ from the original heme deformations. The protein matrix induced similar distortions on the hemes in orientations A and B. Our results suggest that the binding site prefers a particular macrocycle conformation, and a 180 degrees rotation of the heme does not significantly alter the protein's preference for this conformation. The intrinsic rotational strengths of the two Soret transitions, separated according to their polarization in the heme plane, show strong correlations with the ruffling deformation and the average twist angle of the pyrrole rings. The total rotational strength, which includes contributions from the chromophores in the protein, shows a weaker correlation with heme distortions. PMID- 11916425 TI - Detecting the adsorption of dye molecules in homogeneous poly(propylene imine) dendrimer monolayers by surface plasmon resonance sensor. AB - In this work, we studied guest-host interactions between various dye molecules and the fifth-generation poly(propylene imine) (PPI-5) dendrimers in aqueous solutions using a surface plasmon resonance (SPR) sensor. The effect of the properties of guest and host molecules (e.g., charge and shape) and media (e.g., pH and ion strength) on affinity between guest and host molecules was investigated. Based on an immobilized homogeneous monolayer of PPI-5 dendrimer tethered to carboxyl-terminal self-assembled monolayers, the adsorption behavior of a group of dye molecules in PPI-5 was obtained. Results show that the strong affinity of PPI-5 to Rose Bengal and erythrosine B is attributed to the good match in charge and shape between the cavities of the dendrimer and the dye molecules. Maximum adsorption around a pH value of 7 was observed. The kinetic behaviors of different dye molecules in dendrimers were also studied. A fundamental understanding of guest-host interactions in dendrimers will guide the design of new-generation sensors and drug delivery carriers. PMID- 11916426 TI - Breaking bonds by mechanical stress: when do electrons decide for the other side? AB - Using first-principles molecular dynamics, we have simulated reactions that can be induced by mechanical stress in a polymer. We have stretched a small piece of poly(ethylene glycol) (PEG) in water at finite temperature. Both the molecule and the solvent were described quantum mechanically on an equal level. The formation of ions was observed, which corresponds to a heterolytic bond cleavage. We were able to monitor the motion of the electrons during the reactions. Our simulations show that the electron transfer and the breaking of the bond occur almost simultaneously and that both processes are initiated by the approach of a solvent molecule toward the destabilized bond. PMID- 11916427 TI - Intra- and intermolecular topological properties of amino acids: a comparative study of experimental and theoretical results. AB - The charge densities rho(r) of the six amino acids L-Asn.H(2)O, DL-Glu.H(2)O, DL Lys.HCl, DL-Pro.H(2)O, DL-Ser, and DL-Val were determined from high-resolution X ray diffraction experiments at 100 K using synchrotron radiation and area detection (CCD) techniques. Bond topological parameters derived from these densities and from those of six additional amino acids published earlier are compared to each other and to the results of ab initio calculations. Experimental and theoretical properties for each chemically equivalent bond are in a fair agreement, and their variances are of similar magnitude. A noticeable outlier is the positive curvature of the density at the bond critical point, for which no correlation between the experimental and theoretical values can be established. The location of nonbonded valence shell charge concentrations derived from the crystalline densities scatter in a wider range than those obtained for the isolated molecules. PMID- 11916428 TI - The laser-induced blue state of bacteriorhodopsin: mechanistic and color regulatory roles of protein-protein interactions, protein-lipid interactions, and metal ions. AB - In this paper we characterize the mechanistic roles of the crystalline purple membrane (PM) lattice, the earliest bacteriorhodopsin (BR) photocycle intermediates, and divalent cations in the conversion of PM to laser-induced blue membrane (LIBM; lambda(max)= 605 nm) upon irradiation with intense 532 nm pulses by contrasting the photoconversion of PM with that of monomeric BR solubilized in reduced Triton X-100 detergent. Monomeric BR forms a previously unreported colorless monomer photoproduct which lacks a chromophore band in the visible region but manifests a new band centered near 360 nm similar to the 360 nm band in LIBM. The 360 nm band in both LIBM and colorless monomer originates from a Schiff base-reduced retinyl chromophore which remains covalently linked to bacterioopsin. Both the PM-->LIBM and monomer-->colorless monomer photoconversions are mediated by similar biphotonic mechanisms, indicating that the photochemistry is localized within single BR monomers and is not influenced by BR-BR interactions. The excessively large two-photon absorptivities (> or =10(6) cm(4) s molecule(-1) photon(-1)) of these photoconversions, the temporal and spectral characteristics of pulses which generate LIBM in high yield, and an action spectrum for the PM-->LIBM photoconversion all indicate that the PM-->LIBM and Mon-->CMon photoconversions are both mediated by a sequential biphotonic mechanism in which is the intermediate which absorbs the second photon. The purple-->blue color change results from subsequent conformational perturbations of the PM lattice which induce the removal of Ca(2+) and Mg(2+) ions from the PM surface. PMID- 11916429 TI - NMR conformational analysis of antide, a potent antagonist of the gonadotropin releasing hormone. AB - Antide is a decapeptide [(N-Ac-D-Nal(1)-D-Cpa(2)-D-Pal(3)-Ser(4)-Lys(Nic)(5)-D Lys(Nic)(6)-Leu(7)-Ilys(8)-Pro(9)-D-Ala(10)-NH(2)] that acts in vivo as an antagonist of GnRH (gonadotropin-releasing hormone). The conformational behavior of antide has been studied in water, TFE, DMF, and DMSO solutions by means of 2D NMR spectroscopy and molecular dynamics calculations. Antide adopts in aqueous solution a delta-shaped backbone conformation, which is characterized by an irregular turn around residues D-Pal(3)-Ser(4) and by the close spatial proximity of the side chains belonging to D-Nal(1) and Ilys(8) (as many as 17 NOE peaks were detected between these side chains). The side-chain protons of Ilys(8) (especially the H(gamma) ones) present remarkably upfield shifted resonances, because of ring current effects induced by the naphthyl moiety. The upfield shifted resonances of the Ilys(8) H(gamma) hydrogen atoms are strictly characteristic of the water delta-shaped conformation and can be considered as structure markers. The observation of ring current shifted Ilys(8) H(gamma) resonances under different conditions (temperature, pH, solvent) indicates a remarkable stability of the water delta-shaped conformation. Such a conformation is at least partially disrupted in solvent mixtures containing high percentages of organic solvents. TFE can induce a well-defined conformation, which is characterized by an S-shaped backbone conformation. In DMF and DMSO solution, the molecule is basically endowed with a random coil conformation and high fluxionality. Antide fulfills the conformational requirements that are known to play a crucial role in receptor recognition, namely (i) the presence of a turn in the backbone and (ii) the all-trans nature of peptide bonds. In addition, the structural rigidity of antide likely adds a further contribution to the receptor binding affinity. PMID- 11916430 TI - Investigation on the nature of the adsorption sites of pyrrole in alkali exchanged zeolite y by nuclear magnetic resonance in combination with infrared spectroscopy. AB - Multinuclear solid-state NMR and infrared spectroscopy have been applied to investigate the host-guest interactions and the nature of the adsorption sites of pyrrole on alkali-exchanged zeolites Y (LiNaY, NaY, KNaY, and CsNaY). The presence of pyrrole provokes changes in the MAS NMR spectra of (23)Na, (7)Li, and (133)Cs to a degree dependent upon the amount adsorbed. The decrease in the quadrupolar coupling constant for (23)Na as well as the shift for (7)Li and (133)Cs signals are attributed to the interaction of the cation with the pyrrole ring system. The adsorption of pyrrole induces the displacement of cations located at SI' and SII sites toward the supercage to bind the guest molecules. In this way, the distribution of the cations at nonframework sites depends on the amount of adsorbate in the zeolite. At low loadings, pyrrole molecules bind preferentially to more electropositive cation in partially exchanged zeolites Y. Quantitative analysis by (1)H NMR shows that the cation-pyrrole complexes formed possess a stoichiometry of 1:1. The origin of the basic site heterogeneity, evidenced by the presence of several components in the -NH infrared stretching band, is investigated assuming that the heterocycle of pyrrole interacts with cations at SII sites in the supercage and the -NH group forms a hydrogen bond with a basic oxygen atom placed in the framework six-member ring. Making use of the information derived from NMR, it is concluded that the main source of basic site heterogeneity comes from the number of aluminum atoms in the six-member rings of the SII site where the alkaline cation is located. PMID- 11916431 TI - Possible strategies toward the elusive tetraaminodisilene. AB - In this paper we predict, using quantum mechanical calculations, which diaminosilylenes would dimerize to produce strongly bound tetraaminodisilenes, which so far have proven to be elusive. The central idea is that diaminosilylenes with a small singlet-triplet energy difference would dimerize to strongly bonded disilenes. Calculations at the B3LYP/6-311++G(3df,2p)//MP2/6-31G(d) level of theory showed that the energy difference between the singlet and the triplet states (DeltaE(ST)) of diaminosilylenes (R(2)N)(2)Si: (1) strongly depends on (i) the twist angle varphi between the SiN(2) and the R(2)N planes and (ii) the NSiN bond angle alpha at the divalent silicon. DeltaE(ST) decreases with increased twisting (larger varphi) and with widening of alpha. DeltaE(ST) is reduced from 70.7 kcal mol(-1) for planar (H(2)N)(2)Si: (1a) to DeltaE(ST) = 21.7 kcal mol(-1) when varphi is held at 90 degrees. Likewise, the bicyclic diaminosilylenes 1,4 diaza-7-silabicyclo[2.2.1]hepta-7-ylidene and 1,5-diaza-9-silabicyclo[3.3.1]nona 9-ylidene (4a,b), with the nitrogens in the bridgehead positions (varphi = 90 degrees), have DeltaE(ST) values of 45.1 and 38.3 kcal mol(-1), respectively. When dimerized, these silylenes form strongly bonded disilenes 5 (E(dim) = -32.2 kcal mol(-1) (4a) and -41.3 kcal mol(-1) (4b)) with Si=Si bond lengths of 2.239 A (4a) and 2.278 A (4b) (MP2/6-31G(d)//MP2/6-31G(d)). These theoretical predictions pave the way for the synthesis of the first strongly bonded tetraaminodisilene. Due to the steric requirements, also silyl substitution at nitrogen has a significant effect on DeltaE(ST) and [(H(3)Si)(2)N](2)Si: (1d) is predicted to form a stable Si=Si bonded dimer (E(dim)= -24.1 kcal mol(-1)). However, the larger size of the Me(3)Si substituent prevents the formation of a Si=Si bonded dimer of [(Me(3)Si)(2)N](2)Si: (1e). PMID- 11916432 TI - Photogeneration and quenching of reactive oxygen species by urocanic acid. AB - Urocanic acid, UCA, is characterized by two electronic transitions in the UV-B (280-320 nm) which comprise its broad absorption spectrum and give rise to wavelength-dependent isomerization quantum yields. The absorption spectrum of UCA extends into the UV-A (320-400 nm). Given the UV-A component of sunlight is significantly greater than the UV-B component it is hypothesized even weak UV-A photochemistry of UCA could be important for in vivo responses to UV radiation. Degenerate pump-probe experiments performed on t-UCA at several wavelengths in the UV-A reveal an excited-state absorption that undergoes a rapid, approximately 1 ps decay. Photoacoustic experiments performed on both the cis and trans isomers reveal the formation of a long-lived intermediate following UV-A excitation. The efficiency and action spectra for this latter photoactive process are presented and are similar for both isomers of UCA. Cholesterol hydroperoxide assays designed to investigate the nature of the UV-A photoreactivity of t-UCA confirm the production of reactive oxygen species. The bimolecular rate constant for the quenching of singlet oxygen by t-UCA is determined to be 3.5 x 10(6) M(-1) s(-1). Taking into consideration recent theoretical calculations and jet expansion studies of the electronic structure of gas-phase t-UCA, a model is proposed to explain the isomerization and photoreactivity of t-UCA in solution over the UV-A region. PMID- 11916433 TI - The search for bishomoaromatic semibullvalenes and barbaralanes: computational evidence of their identification by UV/Vis and IR spectroscopy and prediction of the existence of a blue bishomoaromatic semibullvalene. AB - Time-dependent B3LYP/6-31G calculations have been performed at the optimized C(2) or C(2v) geometries of several substituted semibullvalenes (1(deloc)) and barbaralanes (2(deloc)), to compare the computed vertical electronic excitation energies with the temperature-dependent, long-wavelength absorptions that have been observed in the UV/vis spectra of some of these compounds by Quast and co workers. The excellent agreement between the calculated vertical excitation energies and the observed values of lambda(max) provides strong support for the identification of the bishomoaromatic species 1(deloc) and 2(deloc) as the source of these absorptions. Furthermore, the CN stretching frequencies, computed for the C(2) geometry of 1,5-dimethyl-2,6-dicyano-4,8-diphenylsemibullvalene (1f(deloc)), fit the low-frequency absorptions seen in the IR spectrum of 1f, thus furnishing independent evidence that bishomoaromatic geometries of semibullvalenes have, in fact, been observed spectroscopically. B3LYP/6-31G calculations predict that 2,6-dicyano-4,8-diphenylsemibullvalene 1c has a C(2) equilibrium geometry (1c(deloc)) and that the long-wavelength UV/vis absorption (lambda(max) = 585 nm) and CN stretching frequencies (2192 and 2194 cm(-1)) computed for 1c(deloc) should serve to identify this bishomoaromatic semibullvalene when it is synthesized. PMID- 11916435 TI - Evaluation of native left ventricular function during mechanical circulatory support: theoretical basis and clinical limitations. AB - Left ventricular function on patients with heart disease is now evaluated by echocardiography, but these dimensional changes are erroneous in the patient supported by left ventricular assist device because of mechanical unloading for the failing heart. Left ventricular end-systolic pressure-volume relationship provides theoretically most reliable left ventricular contractility. Recently, some patients have weaned from the device because of unexpected recovery of myocardial contractility. But it is very important to evaluate the left ventricular function just before the weaning, and to predict the longevity of the recovered function to keep the good quality of life. Current clinical situation in the patients with ventricular assist device, and theoretical limitations to evaluate the recovering myocardium are discussed. PMID- 11916434 TI - Horseradish peroxidase catalyzed nitric oxide formation from hydroxyurea. AB - Hydroxyurea represents an approved treatment for sickle cell anemia and a number of cancers. Chemiluminescence and electron paramagnetic resonance spectroscopic studies show horseradish peroxidase catalyzes the formation of nitric oxide from hydroxyurea in the presence of hydrogen peroxide. Gas chromatographic headspace analysis and infrared spectroscopy also reveal the production of nitrous oxide in this reaction, which provides evidence for nitroxyl, the one-electron reduced form of nitric oxide. These reactions also generate carbon dioxide, ammonia, nitrite, and nitrate. None of these products form within 1 h in the absence of hydrogen peroxide or horseradish peroxidase. Electron paramagnetic resonance spectroscopy and trapping studies show the intermediacy of a nitroxide radical and a C-nitroso species during this reaction. Absorption spectroscopy indicates that both compounds I and II of horseradish peroxidase act as one-electron oxidants of hydroxyurea. Nitroxyl, generated from Angeli's salt, reacts with ferric horseradish peroxidase to produce a ferrous horseradish peroxidase-nitric oxide complex. Electron paramagnetic resonance experiments with a nitric oxide specific trap reveal that horseradish peroxidase is capable of oxidizing nitroxyl to nitric oxide. A mechanistic model that includes the observed nitroxide radical and C-nitroso compound intermediates has been forwarded to explain the observed product distribution. These studies suggest that direct nitric oxide producing reactions of hydroxyurea and peroxidases may contribute to the overall pharmacological properties of this drug. PMID- 11916436 TI - Preclinical evaluation of a new hollow fiber silicone membrane oxygenator for pediatric cardiopulmonary bypass: ex-vivo study. AB - Based on the results of many experimental models, a hollow fiber silicone membrane oxygenator applicable for long-term extracorporeal membrane oxygenation (ECMO) was developed. For further high performance and antithrombogenicity, this preclinical model was modified, and a new improved oxygenator was successfully developed. In addition to ECMO application, the superior biocompatibility of silicone must be advantageous for pediatric cardiopulmonary bypass (CPB). An ex vivo short-term durability test for pediatric CPB was performed using a healthy miniature calf for six hours. Venous blood was drained from the left jugular vein of a calf, passed through the oxygenator and infused into the left carotid artery using a Gyro C1E3 centrifugal pump. For six hours, the O2 and CO2 gas transfer rates were maintained around 90 and 80 ml/min at a blood flow rate of 2 L/min and V/Q=3, respectively. The plasma free hemoglobin was maintained around 5 mg/dl. These data suggest that this newly improved oxygenator has superior efficiency, less blood trauma, and may be suitable for not only long-term ECMO but also pediatric CPB usage. PMID- 11916437 TI - Cytokine responses to myocardial revascularization on cardiopulmonary bypass: intermittent crossclamping versus blood cardioplegic arrest. AB - BACKGROUND: The inflammatory responses to the different myocardial protection approaches have not been thoroughly investigated. We sought to study the cytokine responses to cardiopulmonary bypass (CPB) using the intermittent aortic crossclamping with ventricular fibrillation (IAC) versus blood cardioplegic arrest (BC) techniques. METHODS: Perioperative plasma levels of tumor necrosis factor a (TNF-alpha), interleukins (IL) 6, 8, 10, and cardiac troponin-I (cTnI) were measured serially before surgery, at the end of surgery, and 2, 24, and 48 hours after elective coronary revascularization in 31 patients (IAC: n=15; BC: n=16). RESULTS: Demographics, preoperative status, and number of grafts (IAC: 2.7 +/- 0.6; BC: 3.0 +/- 0.4) were similar between groups. No major complications occurred in either group. The total ischemic time and duration of CPB were shorter in group IAC (17 +/- 5 and 58 +/- 10 min vs 45 +/- 14 and 81 +/- 21 min; both p<0.01). Although the intergroup difference in postoperative cTnI levels was not statistically significant, the release of both TNF-alpha and IL-8 were higher in group IAC than in group BC. However, IL-6 and IL-10 levels were lower after surgery in group IAC. CONCLUSION: Despite the duration of ischemia and CPB being shorter, intermittent aortic crossclamping is associated with an enhanced pro inflammatory but a reduced anti-inflammatory response compared to the cardioplegic arrest technique. Its clinical relevance needs to be further defined. PMID- 11916438 TI - Reoperation for coronary artery disease: devised management for reoperation including digital subtraction angiography. AB - Between December 1990 and August 1999, a consecutive series of 494 patients underwent coronary artery bypass grafting, while 19 (3.8%) patients underwent isolated reoperative coronary bypass grafting. The mean age of 16 males and 3 females who underwent reoperation was 63.4 +/- 6.4 years. The mean interval from the initial operation to reoperation was 50.7 +/- 61.0 months (range 3 days to 16 years). There were neither operative nor in-hospital deaths. Postoperative graft patency was 96.9% in all the bypass grafts and 96.7% in the arterial grafts. Despite the small number of reoperations, the outcomes were favorable due to our devised management: the patients who had received coronary artery bypass grafting underwent intra-aortic digital subtraction angiography 7 days postoperatively to evaluate the early graft patency, which subsequently avoided reoperation 1 to 3 months after the initial operation when the pericardial adhesion was tight; in the primary operation, internal thoracic artery grafts were covered with thymus, and the pericardium was closed for an easy sternal re-entry; and in the reoperation, stenotic grafts were dissected and ligated after aortic cross clamping, all sides of the heart were dissected to obtain a good operative field and to prepare for any cardiac accident, arterial grafts were used to avoid re reoperation, and aprotinin was useful in reducing perioperative bleeding. The present findings suggest that intra-aortic digital subtraction angiography, appropriate preparation for follow-up surgery at the primary operation, and complete revascularization using arterial grafts at reoperation appear to have contributed to the satisfactory outcome in coronary reoperation. PMID- 11916439 TI - Ultrastructural alterations in acquired aortic and mitral valve disease as revealed by scanning and transmission electron microscopical investigations. AB - Scanning and transmission electron microscopical investigations were carried out on explanted human aortic and mitral valves to study the prevalence of hyperplastic and degenerative lesions in acquired valvular dysfunction. Biopsies were taken from 67 aortic and 23 mitral valves. All of the valves examined showed degenerative lesions including a loose binding of the endothelial cells, a partial denudation of the endothelial cover and areas of fibrous hyperplasia surrounded by calcium deposits. Additionally, the formation of various excrescences was detected by means of scanning electron microscopy. Of all excrescences identified, 90% were localized at the free margin of the leaflet, 3% in the subnodular region and 7% in the nodule of Arantius. The ratio of filiform to lamellar forms of hyperplastic lesions was approximately 80% in most of the samples examined. The results presented demonstrate the complex ultrastructural features of surgically explanted human valves showing both degenerative and hyperplastic lesions in the same valve. PMID- 11916440 TI - Retrograde continuous warm blood cardioplegia reduces oxidative stress during coronary artery bypass grafting. AB - Myocardial oxidative stress during retrograde continuous blood cardioplegia (RCBC) was evaluated in 22 patients undergoing elective aortocoronary bypass surgery. The patients were divided into two groups: Group C (n=11) received cold RCBC, and Group W (n=11) received warm RCBC. Myocardial oxidative stress was assessed by measuring the release of oxidized glutathione (GSSG), malondialdehyde (MDA), and myeloperoxidase (MPO) in the coronary sinus plasma before aortic clamping, at 1, 5, and 10 minutes after unclamping. Both the hemodynamic recovery and the creatine kinase MB (CKMB) activity were measured perioperatively until 24 hours after unclamping. In Group C, a significant coronary sinus release of GSSG was found in the early reperfusion period in comparison to Group W. No significant difference in the release of MDA nor MPO was noted in the two groups. The recoveries in the left and right ventricular functions, and the peak CK-MB activity were similar in both groups. In conclusion, warm blood cardioplegia is thus considered to protect the myocardium from ischemia-reperfusion injury better than cold blood cardioplegia under retrograde continuous perfusion. PMID- 11916441 TI - The long-term follow-up results of elective surgical treatment for abdominal aortic aneurysms. AB - This study was aimed at evaluating early and long-term follow-up results of surgical reconstruction of infrarenal abdominal aortic aneurysms (AAA). A consecutive series of 392 patients who underwent elective abdominal aortic repair from 1974 to 2000 was reviewed retrospectively. The mean age was 69.8 years (range 34-90), with 329 males and 63 females. The hospital mortality rate was 3.8% (15/392). Of the 203 patients that died during the follow-up period, 28% (56/203) were due to atherosclerotic diseases and 25% (51/203) were malignancies. The Patients whom underwent AAA surgery associated with ischemic heart disease had a 5-, 10-, and 15-year survival rate of 62%, 30%, and 9%, respectively. On the other hand, a survival rate of those not associated with ischemic heart disease were 71%, 38%, and 16%. The patients associated with aortoiliac occlusive disease in AAA surgery had a 5-, 10-, and 15-year survival rate of 51%, 11%, and 0%; those without aortoiliac occlusive disease had a survival rate of 72%, 43%, and 18%, respectively. There were statistically significant differences between the ischemic heart disease and the non-ischemic heart disease, the aortoiliac occlusive disease and the non-aortoiliac occlusive disease in long-term survival rates respectively. These findings demonstrate that AAA patients associated with ischemic heart disease or aortoiliac occlusive disease are at a higher risk than those with AAA alone. Therefore, AAA patients with aortoiliac occlusive disease and or ischemic heart disease should be managed more intensively before, during and after the operation. PMID- 11916442 TI - An intrapulmonary chondromatous hamartoma penetrating the visceral pleura: report of a case. AB - A 65-year-old woman had been followed up for a hamartoma-like mass in the right upper lobe of the lung since 1995. A follow-up CT scan showed an increase in the size of the lesion and a new lesion next to the old mass, which was highly suspected to be lung cancer. A right upper lobectomy was thus performed in May 2000. The hamartoma penetrated the visceral pleura, and adhered to the mediastinal pleura. Such a growing hamartoma with pleural invasion has so far rarely been previously reported. PMID- 11916443 TI - Three-dimensional CT of thoracic outlet syndrome: report of three cases. AB - Recently, three-dimensional (3-D) imaging has become useful for the diagnosis and treatment of several diseases as computed tomography (CT) imaging technology has advanced. We used three dimensional CT for determination of the narrowing of the thoracic outlet (TO) in three patients with thoracic outlet syndrome (TOS). 3D-CT images are effective for visually identifying the relationships of the structures and planning the operation in TOS. PMID- 11916444 TI - Coronary artery bypass grafting in an adult case with Kawasaki disease. AB - Surgical revascularization for coronary artery lesions secondary to Kawasaki disease has been rarely reported in adult patients. We reported an adult case with few coronary risk factors but with multiple coronary artery aneurysms and obstructive lesions presumably secondary to Kawasaki disease who underwent coronary artery bypass grafting (CABG) with multiple arterial grafts. The postoperative course was uneventful. Because coronary artery sequelae of Kawasaki disease can be a cause of ischemic heart disease even in adults, heightened awareness of this possibility is required for young adults with coronary lesions but without coronary risk factors. PMID- 11916445 TI - Aortic valve replacement in a woman with osteogenesis imperfecta. AB - The mortality rate in cardiac surgery patients with heritable generalized connective tissue disorders, such as Marfan s syndrome and osteogenesis imperfecta, is high due to tissue friability. We describe a successful open heart surgery for repair of aortic regurgitation in a woman with osteogenesis imperfecta (OI). Although tissue friability caused no problems during surgery in this case, it should be kept in mind when operating on patients with OI. PMID- 11916446 TI - Serous fluid leakage without seroma after aortobifemora bypass operation. AB - Perigraft seroma and serous fluid leakage is an uncommon clinicopathologic entity. Serous leaks and/or seroma formation from dacron grafts have been reported in peripheral arterial anastomoses. We present a case of serous fluid leakage without seroma which developed after aorto-bifemoral and femoro-popliteal bypass. PMID- 11916447 TI - Aorto-coronary bypass via the oblique sinus. AB - When the radial artery is used as a conduit for the postero-lateral branches, simple aorto-coronary bypass is sometimes impossible in a hypertrophic heart because of its limited length. Passing the graft through the oblique sinus, and making the proximal anastomosis on the right side of the ascending aorta, the postero-lateral branches can be revascularized with a short graft even for a dilated heart. PMID- 11916448 TI - Growth and production of cholesterol oxidase by alginate-immobilized cells of Rhodococcus equi No. 23. AB - Rhodococcus equi No. 23 was immobilized in calcium alginate. No detrimental effect on the viability of the test organism was observed during the immobilization procedure. Approx. 98% of the cell population originally present in the alginate solution were immobilized in the gel beads. When the cells of an equal volume of the culture, obtained respectively at exponential phase (12 h preculture), late-exponential phase (20 h preculture) or stationary phase (36 h preculture) were immobilized, the gel beads prepared with the stationary-phase culture were found to contain the highest cell population [about 10(8) colony forming units (CFU)/g of beads]. In addition, gel beads, prepared with late exponential-phase culture, exhibited the highest production of cholesterol oxidase (CholOx) after 48 h of incubation. Increasing the bead mass from 3.5 to 14.0 g/100 ml of medium increased CholOx production. However, further increasing the bead mass resulted in a reduction of CholOx production. Furthermore, on the basis of a similar initial cell population, the alginate-immobilized cells of R. equi No. 23 produced a significantly higher amount of CholOx (P<0.05) than did the free cells. PMID- 11916449 TI - A high-stringency proteomics concept aimed for generation of antibodies specific for cDNA-encoded proteins. AB - A novel dual bacterial expression system, designed for high-throughput generation of antibodies specific for cDNA-encoded proteins, is presented. The concept involves parallel expression of cDNA-encoded proteins, in two vector systems, as fusions with two different tags, both enabling single-step affinity purification under denaturing conditions. One of the fusion tags includes a portion with documented immunopotentiating effect to stimulate antibody production, and the generated fusion proteins are used to elicit antibodies. The second fusion protein is used in an immobilized form as an affinity ligand to enrich, from the generated antisera, antibodies with selective reactivity to the cDNA-encoded part. To evaluate the system, five cDNA clones from a mouse testis cDNA library were expressed, and antibodies to these cDNA-encoded proteins were generated, enriched and used in blotting procedures to determine expression patterns for the native proteins corresponding to the cDNAs. The five antibody preparations showed specific reactivity to the corresponding recombinant cDNA-encoded proteins, and three of the five antibodies gave specific staining in Western-blot screening of various cell types and tissue homogenates. When the same five cDNAs were processed and analysed using a single-vector method, antibodies with a more non specific staining were generated. We thus conclude that the presented dual-vector method offers a highly stringent strategy for generation of monospecific polyclonal antibodies. PMID- 11916450 TI - One-step affinity purification of fetuin from fetal bovine serum. AB - Fetuin is a plasma glycoprotein widely distributed in mammals. It has been used as a model protein for structural analyses and investigations into the biological properties of glycoproteins. A convenient one-step procedure for biospecific isolation of fetuin from fetal bovine serum was developed on the basis of wheatgerm agglutinin (WGA) affinity separation. Two different porous supports, a silica-based material and a polymer-based material, were used for the immobilization of WGA. The prepared WGA adsorbents were characterized and process parameters of the affinity separation of fetuin were investigated and optimized. WGA was immobilized on silica and polymer supports with coupling yields of 99.6 and 99.4% respectively and amounts of coupled ligand of 7.9 and 9.2 mg of WGA/ml respectively. It has been shown that the specific capacities for fetuin were 5.1 mg/ml on WGA-silica, 1.8 mg/ml on WGA-polymer and 4.1 mg/ml on WGA-agarose. All three adsorbents proved to be suitable for the biospecific separation of fetuin. The polymer-based WGA adsorbent was successfully applied in the purification of fetuin from fetal bovine serum in a one-step separation process. The identity and purity of the isolated product was verified by SDS/PAGE. Under optimized conditions up to 21.6 mg of fetuin could be isolated from 1 ml of serum. The procedure described was designed to be easily scaled-up for the production of fetuin. PMID- 11916451 TI - Genetic design for facilitated production and recovery of recombinant proteins in Escherichia coli. AB - Genetic strategies have been used for more than two decades to improve bacterial bioprocesses and to simplify recovery procedures. Such strategies include the design of efficient expression vectors and the improvement of bacterial production strains in different ways, e.g. by deletion of protease genes or engineering for overexpression of rare-codon tRNAs, foldases or chaperones. Gene multimerization is another such principle that has proved beneficial to improve production yields. Genetic strategies have furthermore been exploited to facilitate recovery processes by adapting the product for a particular purification principle. In this area, affinity fusions have been commonly used, but other principles, such as modified isoelectric point (pI) or hydrophobic properties have also been successfully investigated. A recent drastic step forward in the use of gene technology to improve recovery processes for recombinant proteins is the introduction of combinatorial protein engineering to generate tailor-made product-specific affinity ligands. This strategy, which allows efficient recovery of a recombinant protein in its native form, is likely to be increasingly used also in industrial-scale bioprocesses, since novel protein ligands have been described that can be sanitized using common industrial cleaning-in-place procedures. The examples presented in this review make it evident that genetic strategies will be of utmost importance in the future for facilitating production and recovery of recombinant proteins. PMID- 11916452 TI - Immobilization of an L-aminoacylase-producing strain of Aspergillus oryzae into gelatin pellets and its application in the resolution of D,L-methionine. AB - The conditions for immobilization of an l-aminoacylase-producing strain of Aspergillus oryzae in gelatin and the enzymic characteristics of the immobilized pellets were studied. The optimal concentrations of gelatin, glutaraldehyde and ethyldiamine and time of immobilization were determined. Scanning electron micrographs reveal the cross-linked structure differences between the native and immobilized pellets. Optimum pH and temperature of the native and immobilized pellets were determined. Effects of ionic strength and substrate concentration on relative activity of the native and immobilized pellets were investigated in detail. The immobilized pellets were more stable over broader temperature and pH ranges. In addition, the immobilized pellets showed stable activity under operational and storage conditions. The immobilized pellets lost about 20% of their initial activity after five cycles of reuse. The results reported in this paper show the potential for using the immobilized A. oryzae pellets to resolve d,l-methionine. PMID- 11916453 TI - A simple fractionation protocol for, and a comprehensive study of the molecular properties of, two major endopolygalacturonases from Aspergillus niger. AB - A comprehensive study on purification and characterization of the two endopolygalacturonases from Aspergillus niger, PG II and PG IV, accounting for 70% of the total polygalacturonase activity, is reported. These enzymes were purified to homogeneity using ion-exchange chromatography and gel filtration. The enzymes had specific activities of 982 and 3750 units/mg, and their molecular masses were 61 and 38 kDa, respectively. The pH optimum of PG II was pH 3.8-4.3 and for PG IV it was between pH 3 and 4.6, and the temperature optima also differed for the enzymes. The enzymes preferred pectic acid as a substrate, cleaving it at random, leading to the release of oligogalacturonides as products. The K(m) values of the two enzymes were found to be 0.12 and 0.72% respectively. The enzymes were rich in hydrophilic amino acids and relatively low in the sulphur-containing amino acids. Both enzymes were rich in beta-structure and differed in their tertiary folding. The tryptophan residues were in a hydrophobic environment. The enzymes differed in their thermal stability; the midpoint of thermal inactivation, T(m), of the two enzymes was found to be 43 degrees C for PG II and 46 degrees C for PG IV. PMID- 11916454 TI - Cultivation of Acetobacter xylinum for bacterial cellulose production in a modified airlift reactor. AB - Acetobacter xylinum for bacterial cellulose production was cultivated in a modified airlift reactor. Better results were obtained from the modified reactor than from a conventional bubble column. After 72 h of cultivation, the final concentration of bacterial cellulose was 7.72 g/l and the productivity was 0.107 g/l per h in the modified airlift reactor. The concentration of bacterial cellulose was about three times higher than that produced in the conventional bubble column. Moreover, the bacterial cellulose produced using the modified reactor formed a unique elliptical pellet (the average diameter was 10 mm), which is different from the fibrous form produced using the stirred-tank reactor. The modified airlift reactor with the suspended bacterial cellulose in pellet form had a higher volumetric oxygen-transfer coefficient and mixing capability than that with bacterial cellulose in fibrous form. The dissolved oxygen in the modified airlift reactor could be maintained above 35% throughout the cultivation. PMID- 11916455 TI - Production of non-alcoholic beer using free and immobilized cells of Saccharomyces cerevisiae deficient in the tricarboxylic acid cycle. AB - Production of non-alcoholic beer using Saccharomyces cerevisiae has been studied. Non-recombinant mutant strains with a defect in the synthesis of tricarboxylic acid-cycle enzymes were used and applied in both free and pectate-immobilized form, using both batch and packed-bed continuous systems. After fermentation, basic parameters of the beer produced by five mutant strains were compared with a standard strain of brewing yeast. Results showed that the beer prepared by mutant yeast cells was characterized by lower levels of total alcohols, with ethanol concentrations between 0.07 and 0.31% (w/w). The organic acids produced, especially lactic acid, in concentrations up to 1.38 g x l(-1) had a strong protective effect on the microbial stability of the final product and thus the usual addition of lactic acid could be omitted. Application of the yeast mutants appears to be a good alternative to the classical methods for the production of non-alcoholic beer. PMID- 11916456 TI - An adventure in biotechnology: the development of haemophilia A therapeutics -- from whole-blood transfusion to recombinant DNA to gene therapy. AB - The past decade has seen an explosion in the number of therapeutic proteins available for a wide spectrum of diseases. Some of these proteins are obtained from human plasma. Examples of these therapeutic proteins are albumin, intravenous immunoglobulins and prothrombin complex concentrates. The majority of new therapeutic proteins are, however, derived via recombinant DNA technology. There are other examples where the first therapeutic preparation was a crude preparation derived from plasma or tissue and where subsequent development has resulted in a recombinant form of the therapeutic protein. This article focuses on the development of therapeutics for the treatment of haemophilia A (deficiency of Factor VIII activity). The progression from crude plasma fractions to monoclonal-purified preparations to the more recent development of therapeutic concentrates via recombinant DNA technology is described in some detail. Finally, the current status of gene therapy for haemophilia A is evaluated. Both technical issues as well as market forces are described, as both have had significant impact on the product-development process. PMID- 11916457 TI - Purification and characterization of alkaline protease from Alcaligenes faecalis. AB - Extracellular alkaline protease from the alkalophilic bacterium Alcaligenes faecalis was purified by a combination of ion-exchange and size-exclusion chromatographic methods, and its properties were examined. The purified enzyme had a specific activity of 563.8 micromol of tyrosine/min per mg of protein and gave a single band on native PAGE and SDS/PAGE with a molecular mass of 67 kDa. Gelatin zymogram also revealed one clear zone of proteolytic activity which corresponded to the band obtained with native PAGE and SDS/PAGE. The enzyme had an optimal pH of 9.0 and exhibited its highest activity at 55 degrees C. The enzyme activity was inhibited by PMSF, suggesting the presence of serine residues at the active site. The enzyme had a K(m) of 1.66 mg/ml and a V(max) of 526 units/min per mg of protein with casein as the substrate. PMID- 11916458 TI - Preventing infection from reusable medical equipment: a systematic review. AB - BACKGROUND: In 2000, the World Health Organization (WHO) had eight sets of conflicting recommendations for decontaminating medical equipment. We conducted a systematic review of observational studies to assist WHO in reconciling the various guidelines. This paper summarises the methods developed and illustrates the results for three procedures--alcohol, bleach and povidone iodine. METHODS: We developed a Medline search strategy and applied inclusion criteria specifying the decontamination procedures of interest and an outcome of microbial destruction for a set of marker organisms. We developed protocols to assess the quality of studies and categorised them according to the reliability of the methods used. Through an iterative process we identified best practice for the decontamination methods and key additional factors required to ensure their effectiveness. We identified 88 published papers for inclusion, describing 135 separate studies of decontamination. RESULTS: For disinfection with alcohol, best practice was identified from 23 studies as an exposure to 70-80% ethanol or isopropanol for at least 5 minutes. Bleach was effective for sterilization at a concentration of 5000 ppm for 5 minutes and for disinfection at 1000 ppm for 10 minutes (33 studies). Povidone iodine was only partially effective for disinfection at a concentration of 1% for 15 minutes (15 studies). CONCLUSIONS: Our findings provide an evidence base for WHO guidelines on decontaminating medical equipment. The results support the recommended use of bleach and show that alcohol could be used more widely than current guidelines suggest, provided best practice is followed. The effectiveness of povidone iodine is uncertain. PMID- 11916460 TI - An ounce (or kilogram) of prevention. PMID- 11916461 TI - Insulin resistance: concepts, controversies, and the role of nutrition. AB - Insulin resistance is a prevalent condition, in which insulin loses its normal physiological action. Since people were first classified as insulin resistant over 60 years ago, one of the main discoveries has been that insulin resistance clusters with other risk factors such as obesity, elevated triglycerides, and low high-density lipoprotein cholesterol, increasing cardiovascular disease risk. Although insulin resistance appears to manifest first in the periphery and then in the liver, other sites, such as the brain and the pancreatic &b.beta-cell, may play pathogenic roles. Factors contributing to insulin resistance at these sites include perturbations in free fatty acids, glucose, and hormone-signalling, some of which may be linked to various genetic polymorphisms. Appropriate nutritional treatment for insulin resistance is controversial. Two main approaches are drawn from diabetes recommendations: i) a high-carbohydrate, low-fat, high-fibre diet emphasizing low glycemic-index foods and ii) sharing calories between monounsaturated fat and complex carbohydrate at the expense of saturated fat. Recent interest in insulin resistance has prompted the development of new guidelines. Promising data have also emerged, showing that a high-carbohydrate, high-fibre, low-fat diet plus exercise programs maintained through intensive counselling can decrease diabetes risk by over 40%. Additional research is required to confirm the sustainability of this approach and sort out the determinants of insulin resistance so that more effective nutritional interventions will result. PMID- 11916463 TI - Parents' and pupils' causal attributions for difficult classroom behaviour. AB - BACKGROUND: Studies of causal attributions for difficult pupil behaviour in schools have tended to focus on the perspectives of teachers. AIMS: The present study examines the causal attributions made by parents for difficult behaviour in classrooms. It reveals the structure of these attributions and serves as a direct comparison to a previous study of pupils' attributions (Miller, Ferguson, & Byrne, 2000) and, more indirectly, with the teacher studies. SAMPLE: The participants were 144 parents (106 mothers and 38 fathers) whose children all attended the same inner city school. METHOD: A questionnaire was constructed following three initial small group interviews with parents to identify the wide range of factors they viewed as potential causes of difficult classroom behaviour. This was then administered to all the parents of children attending the same inner-city primary school. RESULTS: Factor analysis indicated that parents' attributions for misbehaviour were best represented by three factors: (1) 'fairness of teachers' actions', (2) 'pupil vulnerability to peer influences and adverse family circumstances', and (3) 'differentiation of classroom demands and expectations'. While there were no differences between parents in terms of gender or ethnicity, they did see the first two factors as more significant contributors to pupil misbehaviour than the third. CONCLUSION: There is both agreement and conflict between parents' attributions and those made by pupils and by teachers. The implementation of policy initiatives, such as home-school agreements and Pastoral Support Programmes, risk exacerbating tensions and difficulties if these differences in attributions are not acknowledged and acted upon. PMID- 11916462 TI - Key process and organization indicators in the dietetic management of dyslipidemia in Canada. AB - Diet interventions for dyslipidemia can produce clinically relevant changes in lipoprotein levels. To determine whether current nutrition counselling practices are consistent with such interventions, we studied aspects of Canadian dietitians' practice. Respondents to a self-administered mail survey (n=350) described practice for three groups of clients: those without and those with cardiovascular disease counselled through ambulatory care, and those with cardiovascular disease who were hospitalized. The process-of-care factors assessed were time spent in initial and follow-up sessions, diet, anthropometry, blood lipids, physical activity, and social and genetic factors. Organization factors assessed included availability of medical history and laboratory data, and perceived support for counselling services. Initial individual interview times averaged one hour, with 49% to 57% of respondents offering scheduled follow up services versus passive or no follow-up services. Overall, counselling practices were consistent with efficacious interventions, but there was wide variation. This was particularly evident in ambulatory care, where higher percentages of clients received follow-up care when respondents reported multidisciplinary group practice; better access to the medical history, and more frequent assessment of measured body weight, client social support, and laboratory data during follow-up care (all p < 0.01). Health care effectiveness may be improved through changes in the process and organization of services. PMID- 11916464 TI - Assessing orientations to learning to teach. AB - BACKGROUND: An important purpose of teacher education is that student teachers develop and change their existing knowledge on learning and teaching. Research on how student teachers variously engage in this process is scarce. In a previous study of 30 student teachers, we identified five different orientations to learning to teach. AIMS: Our aim was to extend the results of the previous study by developing an instrument to assess orientations to learning to teach at a larger scale. The development and psychometric properties of the instrument are discussed. The results with respect to how student teachers learn are compared to the results of the qualitative study. SAMPLE: Participants in this study were 169 secondary student teachers from three institutes which had all adopted an initial in-service model of learning to teach. METHODS: On the basis of extensive qualitative study, a questionnaire was developed to assess individual differences in learning to teach. Factor-, reliability-, and nonparametric scalability analyses were performed to identify reliable scales. Cluster analysis was used to identify groups of students with similar orientations to learning to teach. RESULTS: Eight scales covering cognitive, regulative and affective aspects of student teachers' learning were identified. Cluster analysis indicates that the instrument discriminates well between student teachers. Four of the five previously found patterns were found again. CONCLUSIONS: The four orientations found in relatively uniform learning environments indicate that student teachers need differential support in their learning. Although the instrument measures individual differences in a reliable way, it is somewhat one-sided in the sense that items representing constructive ways of learning dominate. New items forming a broader range of scales should be created. PMID- 11916465 TI - Creating a scale to measure motivation to achieve academically: linking attitudes and behaviours using Rasch measurement. AB - BACKGROUND: Motivation to Achieve Academically has been used in many educational and other studies in many countries and the large majority has not used an interval level scale based on a good theoretical model in which the items are linked to behaviour. AIMS: One, to create an interval level, unidimensional scale of Motivation, with attitude items linked to behaviour items, based on a conceptual model of Motivation, involving Striving for Excellence (Standards, Goals, Tasks, Effort, Values and Ability), Desire to Learn (Interest, Learning from Others and Responsibility for Learning), and Rewards (Extrinsic, Intrinsic and Social). Two, to analyse its psychometric properties using the Extended Logistic Model of Rasch (Andrich, 1988a, 1988b; Rasch, 1980/1960). Three, to investigate the structure and meaning of the scale. SAMPLE: The convenience person sample was 239 first-year students selected for three special entry programmes at an Australian university. The stem-item sample was initially 45, reduced to 24, that fitted the measurement model to form a valid and reliable scale. METHOD: Based on recent literature, a conceptual model of Motivation was devised and items written in line with the model. Data were collected by self report questionnaire and analysed with the computer program Rasch Unidimensional Measurement Models (RUMM) (Andrich, Lyne, Sheridan, & Luo, 1998). A scale was created in which the Motivation measures were calibrated on the same scale as the item 'difficulties'. RESULTS: Twenty-four Motivation items fitted the model and were 'easier' than their corresponding behaviour items, as conceptualised. They formed an excellent scale in which the proportion of observed variance considered true was 0.93. Items from all aspects of the Motivation model named in the aims above, except Ability and Extrinsic Rewards, fitted the measurement model. CONCLUSION: The Rasch model and the RUMM computing program were very useful in creating a unidimensional, interval level scale of Motivation to achieve academically, with good psychometric properties. PMID- 11916466 TI - Relationship among achievement goal orientations and multidimensional situational motivation in physical education. AB - BACKGROUND: Contemporary research suggests that task and ego achievement goal orientations affect students' intrinsic motivation in physical education. This research has assessed intrinsic motivation as a unidimensional contruct, however, which is inconsistent with the more contemporary postulates of self-determination theory (Deci & Ryan, 1985, 1991) which states that intrinsic motivation is only one type of motivation. To date, research has not addressed whether different types of motivation at the situational level are influenced by the proneness to adopt task or ego involvement. AIMS: To examine the relationship between achievement goal orientations and multidimensional situational motivation in PE. SAMPLE: Middle school children (182 male, 136 female; M age = 13.2 years). METHOD: Responded to questionnaires assessing their dispositional goal orientation (POSQ; Roberts, Treasure, & Balague, 1998) and situational motivation (SIMS; Guay, Vallerand, & Blanchard, 2000) in PE. RESULTS: Task orientation was found to be positively associated with more self-determined types of situational motivation. Ego orientation was weakly related to less self-determined motivation. An extreme group split was conducted to create four goal groups and goal profile analyses conducted. A significant MANOVA was followed by univariate analyses, post hoc comparisons, and calculated effect sizes, which revealed that groups high in task orientation reported more motivationally adaptive responses than groups low in task orientation. CONCLUSIONS: The results suggest that a high level of task orientation singularly or in combination with ego orientation fosters self-determined situational motivation in the context of PE. PMID- 11916467 TI - Knowledge and attitudes about school bullying in trainee teachers. AB - BACKGROUND: Recent research has highlighted the issue of bullying in schools. However, little attention has been paid to the knowledge and beliefs of trainee teachers, who will hold a key position in reducing the problem of bullying in schools. AIMS: To determine (i) trainee teachers' beliefs about some aspects of school bullying; (ii) their recommended strategies for pupils to cope; (iii) their confidence in dealing with bullying issues as future teachers; (iv) the value of specific aspects of training; (v) the importance for these of sex, and primary/secondary training. SAMPLE: This comprised 270 students enrolled on either a one-year postgraduate Certificate in Education (PGCE) course (N =197) or a four-year degree in education (BEd) course (N =73). Age ranged from 18-51 years (mean=28 years). METHODS: Questionnaire completed and returned within a 2-week period. RESULTS: Bullying was seen as an important issue. Some aspects of trainee teachers' knowledge of bullying were accurate, others less so. Telling teachers, and parents, were the strategies most highly recommended to pupils. The great majority were in favour of teacher training courses incorporating information regarding ways of combating bullying. However, they had less confidence about their ability to deal with bullying. Respondents expressed more confidence when dealing with victims rather than bullies and working with the parents of victims rather than the parents of bullies. CONCLUSION: It is important that teacher training programmes incorporate information about school bullying, and are designed to take account of existing knowledge, and areas in which trainee teachers may feel less confident. PMID- 11916468 TI - Cognitive assessment of dyslexic students in higher education. AB - BACKGROUND: Previous studies have shown that the phonological deficits that characterise dyslexia persist into adulthood. There is a growing number of dyslexic students in higher education for whom sensitive diagnostic tests of their reading and reading related difficulties are required. AIMS: The main aim of this study was to compare the cognitive skills of dyslexic students with those of their non-dyslexic peers, and to ascertain the impact of cognitive difficulties on their study skills. A second aim was to produce guidelines for the assessment of dyslexia in higher education. SAMPLE: The performance of 23 dyslexic students was compared with that of a comparison group of 50 students from the same university who did not report a history of reading difficulty. METHOD: Participants completed standardised tests of IQ, reading, spelling and arithmetic and tests tapping phonological processing, verbal fluency and speed of processing. Their performance on a set of study-related tasks including proof reading and precis writing was also assessed and they completed the Brown ADD scales. RESULTS: Although dyslexic students did not differ in general cognitive ability from controls, they had deficits in reading and reading related phonological processes. Discriminant function analyses indicated that dyslexia in adulthood can be confirmed with 95% accuracy using only four tests: spelling, nonword reading, digit span and writing speed. CONCLUSIONS: The study highlighted the difficulties of dyslexic adults. The persisting difficulties of dyslexic students that affect their study skills need to be recognised by HE institutions so that appropriate support programmes can be put in place. PMID- 11916469 TI - Subthalamic-pallidal interactions are critical in determining normal and abnormal functioning of the basal ganglia. AB - The subthalamic nucleus (STN) and external globus pallidus (GP) form a recurrent excitatory-inhibitory interaction within the basal ganglia. Through a computational model of these interactions we show that, under the influence of appropriate external input, the two nuclei can be switched between states of high and low activity or can generate oscillations consisting of bursts of high frequency activity repeated at a low rate. It is further demonstrated from the model that the generation of the repetitive bursting behaviour is favoured by increased inhibition of the GP, which is a condition indicated in Parkinson's disease. Paradoxically, increased striatal inhibition of the GP is predicted to cause an increase rather than a decrease in its mean firing rate. These behaviours, arising from a biologically inspired computational model of the STN GP interaction, have important consequences for basal ganglia function and dysfunction. PMID- 11916470 TI - Life-history theory, fertility and reproductive success in humans. AB - According to life-history theory, any organism that maximizes fitness will face a trade-off between female fertility and offspring survivorship. This trade-off has been demonstrated in a variety of species, but explicit tests in humans have found a positive linear relationship between fitness and fertility. The failure to demonstrate a maximum beyond which additional births cease to enhance fitness is potentially at odds with the view that human fertility behaviour is currently adaptive. Here we report, to our knowledge, the first clear evidence for the predicted nonlinear relationship between female fertility and reproductive success in a human population, the Dogon of Mali, West Africa. The predicted maximum reproductive success of 4.1+/-0.3 surviving offspring was attained at a fertility of 10.5 births. Eighty-three per cent of the women achieved a lifetime fertility level (7-13 births) for which the predicted mean reproductive success was within the confidence limits (3.4 to 4.8) for reproductive success at the optimal fertility level. Child mortality, rather than fertility, was the primary determinant of fitness. Since the Dogon people are farmers, our results do not support the assumptions that: (i) contemporary foragers behave more adaptively than agriculturalists, and (ii) that adaptive fertility behaviour ceased with the Neolithic revolution some 9000 years ago. We also present a new method that avoids common biases in measures of reproductive success. PMID- 11916471 TI - Short-term studies underestimate 30-generation changes in a butterfly metapopulation. AB - Most studies of rare and endangered species are based on work carried out within one generation, or over one to a few generations of the study organism. We report the results of a study that spans 30 generations (years) of the entire natural range of a butterfly race that is endemic to 35 km(2) of north Wales, UK. Short term studies (surveys in single years and dynamics over 4 years) of this system led to the prediction that the regional distribution would be quite stable, and that colonization and extinction dynamics would be relatively unimportant. However, a longer-term study revealed unexpectedly high levels of population turnover (local extinction and colonization), affecting 18 out of the 20 patches that were occupied at any time during the period. Modelling the system (using the 'incidence function model' (IFM) for metapopulations) also showed higher levels of colonization and extinction with increasing duration of the study. The longer term dynamics observed in this system can be compared, at a metapopulation level, with the increased levels of variation observed with increasing time that have been observed in single populations. Long-term changes may arise from local changes in the environment that make individual patches more or less suitable for the butterfly, or from unusual colonization or extinction events that take metapopulations into alternative states. One implication is that metapopulation and population viability analyses based on studies that cover only a few animal or plant generations may underestimate extinction threats. PMID- 11916472 TI - The ecology of extinction: molluscan feeding and faunal turnover in the Caribbean Neogene. AB - Molluscan faunal turnover in the Plio-Pleistocene of the tropical western Atlantic has been attributed to drops in temperature or primary productivity, but these competing hypotheses have not been assessed ecologically. To test these alternatives, we compiled data on changing molluscan life habits and trophic composition over 12 million years derived from 463 newly made collections from the southwestern Caribbean. Shelf ecosystems have altered markedly in trophic structure since the Late Pliocene. Predatory gastropods and suspension-feeding bivalves declined significantly in abundance, but not in diversity, and reef dwellers became common. By contrast, all other ecological life habits remained remarkably stable. Food-web changes strongly support the hypothesis that declining regional nutrient supply had an increasing impact on regional macroecology, culminating in a faunal turnover. PMID- 11916473 TI - Determinants of asynchronous processing in vision. AB - When a stimulus oscillates in both colour and direction of motion, changes in colour must lag behind those in direction if they are to be seen as concurrent. It has been argued that this lag is the consequence of asynchronous visual processing, with colour being processed more rapidly than motion. This proposal is contentious: it has been criticized on the basis that the time-course of cortical activity may not correlate directly with that of perceptual experience. Here, we demonstrate that the extent of the apparent asynchrony can vary according to the prevailing stimulus conditions. The apparent asynchrony is greatest if the stimulus is composed of opponent directions of motion and is reduced if the angular difference between the directions is reduced. This pattern of results suggests that asynchronous neural activity arises, in part, as a consequence of differential levels of inhibition within relatively independent cortical structures. PMID- 11916474 TI - The evolution of risky behaviour in the presence of a sexually transmitted disease. AB - Sexually transmitted diseases (STDs) are widespread in nature, often sterilizing their hosts or causing other pathogenic effects. Despite this, there is a widespread occurrence of behaviours that are likely to increase the risk to an individual of contracting an STD. Here, we examine the evolution of behaviours such as promiscuity or mate choice that increase the risk of contracting an STD, but also provide a fitness benefit. As might be expected, the balance between risk and fitness benefit defines the optimal strategy, but this relationship is not straightforward. In particular, we often predict the coexistence of highly risky and highly risk-averse individuals. Surprisingly, very safe strategists that only suffer a small cost will tend to coexist with highly risky strategists rather than outcompete them as might have been expected. Rather than selecting for monogamy or for reduced mate choice, therefore, the presence of an STD may often lead to variability in either promiscuity or mate choice. PMID- 11916475 TI - Mutator dynamics in fluctuating environments. AB - Populations with high mutation rates (mutator clones) are being found in increasing numbers of species, and a clear link is being established between the presence of mutator clones and drug resistance. Mutator clones exist despite the fact that in a constant environment most mutations are deleterious, with the spontaneous mutation rate generally held at a low value. This implies that mutator clones have an important role in the adaptation of organisms to changing environments. Our study examines how mutator dynamics vary according to the frequency of environmental fluctuations. Although recent studies have considered a single environmental switch, here we investigate mutator dynamics in a regularly varying environment, seeking to mimic conditions present, for example, under certain drug or pesticide regimes. Our model provides four significant new insights. First, the results demonstrate that mutators are most prevalent under intermediate rates of environmental change. When the environment oscillates more rapidly, mutators are unable to provide sufficient adaptability to keep pace with the frequent changes in selection pressure and, instead, a population of 'environmental generalists' dominates. Second, our findings reveal that mutator dynamics may be complex, exhibiting limit cycles and chaos. Third, we demonstrate that when each beneficial mutation provides a greater gain in fitness, mutators achieve higher densities in more rapidly fluctuating environments. Fourth, we find that mutators of intermediate strength reach higher densities than very weak or strong mutators. PMID- 11916476 TI - Species boundaries, populations and colour morphs in the coral reef three-spot damselfish (Dascyllus trimaculatus) species complex. AB - Coloration patterns of tropical reef fishes is commonly used for taxonomic purposes, yet few studies have focused on the relationship between species boundaries and coloration types. The three-spot damselfish (Dascyllus trimaculatus) species complex comprises four species that vary both in geographical ranges and colour patterns making them an ideal model to study these relationships. We analysed the mitochondrial control region of 122 individuals from all four species collected from 13 localities. Individuals from two species (Dascyllus albisella and D. strasburgi) grouped into monophyletic clades, while the two other species (D. trimaculatus and D. auripinnis) were found to be paraphyletic. Coloration patterns were therefore not found to be good predictors of genetic isolation. In contrast, geographical origin was always consistent with the observed genetic pattern. PMID- 11916477 TI - Sperm mobility: mechanisms of fertilizing efficiency, genetic variation and phenotypic relationship with male status in the domestic fowl, Gallus gallus domesticus. AB - When females are sexually promiscuous, sexual selection continues after insemination through sperm competition and cryptic female choice, and male traits conveying an advantage in competitive fertilization are selected for. Although individual male and ejaculate traits are known to influence paternity in a competitive scenario, multiple mechanisms co-occur and interact to determine paternity. The way in which different traits interact with each other and the mechanisms through which their heritability is maintained despite selection remain unresolved. In the promiscuous fowl, paternity is determined by the number of sperm inseminated into a female, which is mediated by male social dominance, and by the quality of the sperm inseminated, measured as sperm mobility. Here we show that: (i) the number of sperm inseminated determines how many sperm reach the female sperm-storage sites, and that sperm mobility mediates the fertilizing efficiency of inseminated sperm, mainly by determining the rate at which sperm are released from the female storage sites, (ii) like social status, sperm mobility is heritable, and (iii) subdominant males are significantly more likely to have higher sperm mobility than dominant males. This study indicates that although the functions of social status and sperm mobility are highly interdependent, the lack of phenotypic integration of these traits may maintain the variability of male fitness and heritability of fertilizing efficiency. PMID- 11916478 TI - An analysis of dinosaurian biogeography: evidence for the existence of vicariance and dispersal patterns caused by geological events. AB - As the supercontinent Pangaea fragmented during the Mesozoic era, dinosaur faunas were divided into isolated populations living on separate continents. It has been predicted, therefore, that dinosaur distributions should display a branching ('vicariance') pattern that corresponds with the sequence and timing of continental break-up. Several recent studies, however, minimize the importance of plate tectonics and instead suggest that dispersal and regional extinction were the main controls on dinosaur biogeography. Here, in order to test the vicariance hypothesis, we apply a cladistic biogeographical method to a large dataset on dinosaur relationships and distributions. We also introduce a methodological refinement termed 'time-slicing', which is shown to be a key step in the detection of ancient biogeographical patterns. These analyses reveal biogeographical patterns that closely correlate with palaeogeography. The results provide the first statistically robust evidence that, from Middle Jurassic to mid Cretaceous times, tectonic events had a major role in determining where and when particular dinosaur groups flourished. The fact that evolutionary trees for extinct organisms preserve such distribution patterns opens up a new and fruitful direction for palaeobiogeographical research. PMID- 11916479 TI - Deleterious Wolbachia in the ant Formica truncorum. AB - Wolbachia is a maternally inherited bacterium that may manipulate the reproduction of its arthropod hosts. In insects, it is known to lead to inviable matings, cause asexual reproduction or kill male offspring, all to its own benefit, but to the detriment of its host. In social Hymenoptera, Wolbachia occurs widely, but little is known about its fitness effects. We report on a Wolbachia infection in the wood ant Formica truncorum, and evaluate whether it influences reproductive patterns. All 33 colonies of the study population were infected, suggesting that Wolbachia infection is at, or close to, fixation. Interestingly, in colonies with fewer infected workers, significantly more sexuals are produced, indicating that Wolbachia has deleterious effects in this species. In addition, adult workers are shown to have significantly lower infection rates (45%) than worker pupae (87%) or virgin queens (94%), suggesting that workers lose their infection over life. Clearance of Wolbachia infection has, to our knowledge, never been shown in any other natural system, but we argue that it may, in this case, represent an adaptive strategy to reduce colony load. The cause of fixation requires further study, but our data strongly suggest that Wolbachia has no influence on the sex ratio in this species. PMID- 11916480 TI - Pollinator-mediated selection on flower-tube length in a hawkmoth-pollinated Gladiolus (Iridaceae). AB - Darwin's mechanistic model whereby selection favours plants with flower tubes that exceed the tongue length of the primary pollinator, was tested using unmanipulated plants of the hawkmoth-pollinated South African iris, Gladiolus longicollis. The study population was characterized by exceptionally large phenotypic variation in flower-tube length (range 56-129 mm). Directional selection on tube length was revealed by a significant positive relationship between this trait and both fruit and seed set. Selection was attributed to the effect of tube length on pollen receipt, as supplemental hand pollinations showed fruit and seed set in the population to be pollen limited. Indirect selection on tube length may also occur through the correlation of this trait with inflorescence height, although direct selection on the latter trait was significant only for seed set. The main pollinators at the study site were individuals of the large hawkmoth Agrius convolvuli that had tongue lengths of 85 135 mm. Other hawkmoths had tongues that were much too short to reach the nectar in G. longicollis flowers and seldom carried pollen of G. longicollis. Flowers with tubes shorter than the tongues of A. convolvuli are apparently not effectively pollinated because stigmas do not contact the moth's head effectively. This study demonstrates that selection may occur among plants with natural phenotypic variation in flower-tube length, and supports Darwin's model of pollinator-mediated selection. PMID- 11916481 TI - Evolution of density- and patch-size-dependent dispersal rates. AB - Based on a marginal value approach, we derive a nonlinear expression for evolutionarily stable (ES) dispersal rates in a metapopulation with global dispersal. For the general case of density-dependent population growth, our analysis shows that individual dispersal rates should decrease with patch capacity and-beyond a certain threshold-increase with population density. We performed a number of spatially explicit, individual-based simulation experiments to test these predictions and to explore further the relevance of variation in the rate of population increase, density dependence, environmental fluctuations and dispersal mortality on the evolution of dispersal rates. They confirm the predictions of our analytical approach. In addition, they show that dispersal rates in metapopulations mostly depend on dispersal mortality and inter-patch variation in population density. The latter is dominantly driven by environmental fluctuations and the rate of population increase. These conclusions are not altered by the introduction of neighbourhood dispersal. With patch capacities in the order of 100 individuals, kin competition seems to be of negligible importance for ES dispersal rates except when overall dispersal rates are low. PMID- 11916482 TI - Cyclic variation in seasonal recruitment and the evolution of the seasonal decline in Ural owl clutch size. AB - Plastic life-history traits can be viewed as adaptive responses to environmental conditions, described by a reaction norm. In birds, the decline in clutch size with advancing laying date has been viewed as a reaction norm in response to the parent's own (somatic or local environmental) condition and the seasonal decline in its offspring's reproductive value. Theory predicts that differences in the seasonal recruitment are mirrored in the seasonal decrease in clutch size. We tested this prediction in the Ural owl. The owl's main prey, voles, show a cycle of low, increase and peak phases. Recruitment probability had a humped distribution in both increase and peak phases. Average recruitment probability was two to three times higher in the increase phase and declined faster in the latter part of the season when compared with the peak phase. Clutch size decreased twice as steep in the peak (0.1 eggs day-1) as in the increase phase (0.05 eggs day-1). This result appears to refute theoretical predictions of seasonal clutch size declines. However, a re-examination of current theory shows that the predictions of modelling are less robust to details of seasonal condition accumulation in birds than originally thought. The observed pattern can be predicted, assuming specifically shaped seasonal increases in condition across individuals. PMID- 11916483 TI - Critical issues in gene therapy for neurologic disease. AB - Gene therapy for the nervous system is a newly emerging field with special issues related to modes of delivery, potential toxicity, and realistic expectations for treatment of this vital and highly complex tissue. This review focuses on the potential for gene delivery to the brain, as well as possible risks and benefits of these procedures. This includes discussion of appropriate vectors, such as adeno-associated virus, lentivirus, gutless adenovirus, and herpes simplex virus hybrid amplicons, and cell vehicles, such as neuroprogenitor cells. Routes of delivery for focal and global diseases are enumerated, including use of migratory cells, facilitation of vascular delivery across the blood-brain barrier, cerebrospinal fluid delivery, and convection injection. Attention is given to examples of diseases falling into different etiologic types: metabolic deficiency states, including Canavan disease and lysosomal storage disorders; and degenerative conditions, including Parkinson's disease and other neurodegenerative conditions. PMID- 11916484 TI - Dopaminergic cell loss induced by human A30P alpha-synuclein gene transfer to the rat substantia nigra. AB - Somatic cell gene transfer was used to express a mutant form of alpha-synuclein (alpha-syn) that is associated with Parkinson's disease (PD) in the rat substantia nigra (SN), a brain region that, in humans, degenerates during PD. DNA encoding the A30P mutant of human alpha-syn linked to familial PD was incorporated into an adeno-associated virus vector, which was injected into the adult rat midbrain. The cytomegalovirus/chicken beta-actin promoter was used to drive transgene expression. Over a 1-year time course, this treatment produced three significant features relevant to PD: (1) accumulation of alpha-syn in SN neuron perikarya, (2) Lewy-like dystrophic neurites in the SN and the striatum, and (3) a 53% loss of SN dopamine neurons. However, motor dysfunction was not found in either rotational or rotating rod testing. The lack of behavioral deficits, despite the significant cell loss, may reflect pathogenesis similar to that of PD, where greater than 50% losses occur before motor behavior is affected. PMID- 11916485 TI - Effective gene transfer to human melanomas via integrin-targeted adenoviral vectors. AB - The utility of recombinant adenoviral vectors (Adv) for gene therapy is limited by their low transduction efficiency and lack of specificity for target cells. The low transduction efficiency is often recognized as due to deficiency of the primary adenoviral receptor, the coxsackievirus-adenovirus receptor (CAR). In this paper, studies of CAR levels on human melanoma cell lines confirmed that low transduction efficiency was closely related to deficiency of the adenoviral receptor. To achieve CAR-independent gene transfer via Adv, we modified viral tropism via genetic alteration of the adenovirus type 5 (Ad5) fiber protein. Insertion of an Arg-Gly-Asp (RGD)-containing peptide in the HI loop of the fiber knob domain allowed the virus to use an alternative receptor, the integrin receptor, during the cell entry process. With this modified vector (Adv-F/RGD) transduction was increased 5- to 96-fold relative to a vector containing wild type fiber (Adv-F/wt) in five human melanoma cells expressing integrins of the alpha(v)beta(3), alpha(v)beta(5) class, which are recognized by the RGD peptide motif. In contrast, no significant difference in transduction efficiency between Adv-F/RGD and Adv-F/wt was observed in 293 cells, which show high-level expression of CAR. In this study, we attempted to apply Adv-F/RGD for gene therapy for malignant melanoma. At the same multiplicity of infection, melanoma cells infected with Adv-F/RGD carrying human interleukin 2 (AxCAhIL2-F/RGD) produced a higher level of cytokine than cells infected with AxCAhIL2-F/wt. Treatment by intratumoral injection of AxCAhIL2-F/RGD was more effective than intratumoral injection of AxCAhIL2-F/wt in regressing tumors in a melanoma xenograft model. These data suggest that integrin-targeted adenoviral vectors may be a powerful tool in gene therapy for CAR-deficient melanomas. PMID- 11916486 TI - Replication-selective herpes simplex virus type 1 mutant therapy of cervical cancer is enhanced by low-dose radiation. AB - Herpes simplex virus type 1 (HSV-1)-based oncolytic treatment is a promising therapeutic approach for malignancy. Recombinant strains of HSV-1 containing mutations in the ICP 34.5 protein have been shown to replicate preferentially in rapidly proliferating malignant cells, resulting in a direct cytolytic effect. We assessed the efficacy of multimutated HSV-1 strains on human cervical cancer, and then used these viruses in combination with radiation therapy, the standard treatment for cervical cancer. The HSV-1 mutants 4009, 7020, 3616, and G207 induced significant lysis of three established human cervical cancer cell lines in vitro in a dose-dependent manner. G207 intratumoral treatment of established subcutaneous C33a tumors in severe combined immunodeficient (SCID) mice significantly reduced tumor burden by 50%. Weekly and triweekly treatments improved efficacy and inhibited flank tumor growth in an administration frequency dependent manner without toxicity. Combination therapy of a low dose of radiation (1.5 or 3 Gy) and replication-selective HSV mutants infection exhibited increased antitumor effects against cervical cancer cells in vitro. The in vivo effect of G207 combined with low-dose radiation was studied in Me180 xenografts in athymic mice. Treatment of established Me180 tumor nodules with 3 Gy followed by intratumoral G207 administration greatly improved efficacy, resulting in 42% complete eradication of tumor. In conclusion, single and multiple intratumoral injections of G207 significantly reduced tumor burden in xenogeneic models of cervical cancer, and the addition of low-dose radiation further potentiated the effect. These results suggest that replication-selective HSV-1 mutants may be potent oncolytic agents for the treatment of cervical cancer. PMID- 11916487 TI - Reovirus oncolysis of human breast cancer. AB - We have previously shown that human reovirus replication is restricted to cells with an activated Ras pathway, and that reovirus could be used as an effective oncolytic agent against human glioblastoma xenografts. This study examines in more detail the feasibility of reovirus as a therapeutic for breast cancer, a subset of cancer in which direct activating mutations in the ras proto-oncogene are rare, and yet where unregulated stimulation of Ras signaling pathways is important in the pathogenesis of the disease. We demonstrate herein the efficient lysis of breast tumor-derived cell lines by the virus, whereas normal breast cells resist infection in vitro. In vivo studies of reovirus breast cancer therapy reveal that viral administration could cause tumor regression in an MDA MB-435S mammary fat pad model in severe combined immunodeficient mice. Reovirus could also effect regression of tumors remote from the injection site in an MDA MB-468 bilateral tumor model, raising the possibility of systemic therapy of breast cancer by the oncolytic agent. Finally, the ability of reovirus to act against primary breast tumor samples not propagated as cell lines was evaluated; we found that reovirus could indeed replicate in ex vivo surgical specimens. Overall, reovirus shows promise as a potential breast cancer therapeutic. PMID- 11916488 TI - Role of hepatocyte direct hyperplasia in lentivirus-mediated liver transduction in vivo. AB - Lentiviral vectors have been used for gene transfer into the liver, but the ability of these vectors to efficiently transduce quiescent hepatocytes remains controversial. Regardless, lentivirus-mediated gene transfer is greatly enhanced when delivered during hepatocellular cycling. For this reason, the present study was designed to determine the role of hepatocyte proliferation in the enhancement of lentiviral transduction by using three different modes of liver regeneration: (1) compensatory regeneration stimulated by two-thirds partial hepatectomy, (2) direct hyperplasia after intragastric administration of the primary mitogen 1,4 bis[2-(3,5-dichloropyridyloxy)] benzene (TCPOBOP), and (3) a combination of modes 1 and 2. Vesicular stomatitis virus glycoprotein (VSV-G)-pseudotyped lentiviral vector expressing beta-galactosidase was administered to mice via the peripheral circulation after a regeneration stimulus. Gene transfer as measured by 5-bromo-4 chloro-3-indolyl-beta-D-galactoside (X-Gal) staining showed 30-fold higher levels of liver transduction in groups 1 and 2 as compared with the non-liver manipulated control group (p < 0.005). The combination of TCPOBOP and partial hepatectomy (group 3) resulted in an ~80-fold increase in transduction efficiency compared with the control animals. The enhanced transduction was consistent with higher levels of hepatocellular proliferation observed in animals that received both treatments compared with either single treatment alone. Importantly, the hepatocytes were the predominant cell type transduced, although transgene expression was observed in a low number of nonparenchymal cells regardless of which liver stimulus was received. Biodistribution studies confirmed that most of the gene transfer was limited to the liver and spleen. Taken together, this study suggests that disease-induced cellular proliferation in the liver will enhance the utility of this vector in treating diseases such as viral hepatitis, liver cirrhosis, and cancer. PMID- 11916489 TI - Highly efficient and specific gene transfer to Purkinje cells in vivo using a herpes simplex virus I amplicon. AB - The transduction of cerebellar neurons in vivo with herpes simplex virus 1 (HSV 1) amplicon carrying the lacZ gene has been investigated after injection of the vector in the cerebellar cortex, ventricles, and inferior olive of adult rats. Injection into the cerebellar cortex resulted in transduction of Purkinje cells near the needle tract and injection into the ventricles yielded no transduced neurons. In contrast, high transduction efficiency was achieved by vector injection into the inferior olive, resulting in one of three positive Purkinje cells all over the ipsilateral and contralateral cerebellar hemispheres. Because neurons in the deep cerebellar nuclei are also transduced, we suggest that the vector is delivered from the inferior olive to the cerebellar nuclei and then to Purkinje cells by retrograde axonal transport. Expression of the lacZ gene within Purkinje cells was surprisingly persistent and was maintained at the same level for at least 40 days. Importantly, no signs of either toxicity or inflammation were observed in the cerebellum after vector injection, except for the borders of the needle tract where some reactive astrocytes were detected. Indeed, motor coordination of treated animals was entirely normal, as assessed by the rota-rod test. These results demonstrate that HSV-1 amplicon vectors can effect safe and stable transgene expression in Purkinje cells in vivo, raising the possibility of using these vectors for long-term gene therapy of human cerebellar disorders. PMID- 11916490 TI - Clinical protocol. Liposomal gene therapy with the herpes simplex thymidine kinase gene/ganciclovir system for the treatment of glioblastoma multiforme. PMID- 11916492 TI - Increasing prevalence of HSV-2 points to need for more effective prevention strategies. PMID- 11916494 TI - Condom use and the prevention of genital herpes acquisition. AB - Genital herpes establishes a lifelong infection associated with significant morbidity and mortality. In contrast to other sexually transmitted infections (STIs), herpes simplex virus type 2 (HSV-2) may be transmitted to sexual partners many years after initial infection and at times when the source partner may be asymptomatic. Consequently, HSV-2 has become one of the most common STIs worldwide. Current therapies suppress but do not eradicate infection. While many studies have examined the degree of protection offered through the use of condoms against HSV-2 acquisition, findings have been either difficult to interpret or inconsistent. However, the body of evidence supports the efficacy of condoms in preventing HSV-2 infection among women. More data are required for HSV-2 prevention in men. The infrequent use of condoms during pregnancy, coupled with the high incidence of complications associated with HSV-2 acquisition at this time, warrants efforts to improve condom use among male partners of pregnant women at risk of HSV-2 infection. PMID- 11916495 TI - The clinical management of recurrent genital herpes: current issues and future prospects. AB - This article reports the proceedings of an expert panel discussion on current clinical management practices for the treatment of recurrent genital herpes. The panel reviewed the effectiveness of primary and specialist care settings in the UK and USA and identified the principal clinical needs of patients with recurrent genital herpes. The ideal alternative to daily suppressive nucleoside analogue therapy is a treatment with long-term impact on the natural history and prognosis of recurrent genital herpes. The potential of resiquimod, an immune response modifier, to resolve this unmet need was examined. Resiquimod is reported to delay the onset of recurrent genital herpes symptoms in patients in a Phase II clinical trial. While awaiting clinical confirmation of this new development in immunotherapy, the panel concluded with two recommendations: a reassessment of recurrent genital herpes management in terms of physician education of patients, and the promotion of a positive patient-physician relationship in the approach to treatment. PMID- 11916496 TI - Opinion article: cytomegalovirus is a risk factor in atherogenesis. AB - Whether or not infectious agents are involved in the pathogenesis of atherosclerosis has been a matter of discussion for the past 2 decades. Although there is no definite proof of a causal role of human cytomegalovirus in atherogenesis, a body of knowledge supports the concept that this virus is involved in the development of atherosclerotic lesions. This review assesses the most important data published in support of this hypothesis. PMID- 11916493 TI - Microbicides for preventing transmission of genital herpes. AB - Available technologies for preventing sexual transmission of genital herpes infection are limited. This article focuses on the ongoing development of a new technology, topical microbicides, for preventing sexually transmitted infection. Recent data evaluating detergent-based spermicides as potential microbicides are reviewed. The first generation of broad-spectrum, non-detergent microbicides that are currently in clinical development, including the sulphated polymer-based inhibitors and acid buffers, are discussed. Finally, the potential of monoclonal antibodies as an example of a specific microbicide in late pre-clinical development is considered. PMID- 11916497 TI - Case study: type-specific HSV serology and the correct diagnosis of first-episode genital herpes during pregnancy. AB - It is now known that the physical presentation of genital herpes simplex (HSV) infection can be misleading in making the diagnosis of genital herpes. An incorrect diagnosis can be particularly damaging in pregnancy where it may result in extended exposure of the fetus to antiviral agents, an inappropriate route and timing of delivery and a significant increase in fetal exposure to HSV during labour and delivery. Case 1 describes a 32-year-old woman at 30 weeks in her first pregnancy who had the appearance and clinical course typically ascribed to primary genital HSV infection. In contrast, Case 2, a 24-year-old woman at 34 weeks' gestation, had the physical appearance of a recurrent episode. Type specific serological testing revealed that what Case 1 was actually experiencing was the first symptomatic reactivation of genital herpes, whereas Case 2 had a true primary genital HSV-2 infection that was accompanied by minimal symptoms. Had serology testing not been available, Case 1 would probably have delivered unnecessarily by Caesarean section, and Case 2 would have been managed as a recurrent infection and allowed to deliver vaginally with potentially disastrous results. These cases illustrate the usefulness of a type-specific serology in diagnosing genital herpes in pregnant women. PMID- 11916498 TI - Comprehensive molecular cytogenetic investigation of chromosomal abnormalities in human medulloblastoma cell lines and xenograft. AB - Cell lines and xenografts derived from medulloblastomas are useful tools to investigate the chromosomal changes in these tumors. Here we used G-banding, fluorescence in situ hybridization (FISH), spectral karyotyping (SKY), and comparative genomic hybridization to study 4 medulloblastoma cell lines and 1 xenograft. Cell line D-425 Med had a relatively simple karyotype, with a terminal deletion of 10q and amplification of MYC in double-minutes (dmins). FISH demonstrated that an apparent isochromosome (17q) by routine karyotyping was actually an unbalanced translocation between 2 copies of chromosome 17. Cell line D-556 Med also had a simple near-diploid stemline with an unbalanced 1;13 translocation resulting in a gain of 1q, an isochromosome (17q), and dmins. These findings were initially described using routine G-banded preparations, and FISH showed that the dmins were an amplification of MYC and the i(17q) was an isodicentric 17q chromosome. The other finding was confirmed by FISH, SKY, and comparative genomic hybridization. Cell lines D-721 Med and D-581 Med had complex karyotypic patterns that could be completely characterized only when FISH and SKY were used. Xenograft D-690 Med also had a complex pattern that FISH and SKY were helpful in completely elucidating. Interestingly, balanced reciprocal translocations were seen as well as complicated unbalanced translocations and marker chromosomes. Comparative genomic hybridization demonstrated only a deletion of 10q22-10q24, supporting the idea that despite the complexity of the chromosomal rearrangements, minimal alterations in the overall chromosomal content had occurred. This study demonstrates that routine cytogenetic preparations are adequate to describe chromosomal abnormalities in occasional medulloblastoma samples, but a broader spectrum of molecular cytogenetic methods is required to completely analyze most of these tumor samples. PMID- 11916499 TI - A chromosomal region 7p11.2 transcript map: its development and application to the study of EGFR amplicons in glioblastoma. AB - Cumulative information available about the organization of amplified chromosomal regions in human tumors suggests that the amplification repeat units, or amplicons, can be of a simple or complex nature. For the former, amplified regions generally retain their native chromosomal configuration and involve a single amplification target sequence. For complex amplicons, amplified DNAs usually undergo substantial reorganization relative to the normal chromosomal regions from which they evolve, and the regions subject to amplification may contain multiple target sequences. Previous efforts to characterize the 7p11.2 epidermal growth factor receptor ) amplicon in glioblastoma have relied primarily on the use of markers positioned by linkage analysis and/or radiation hybrid mapping, both of which are known to have the potential for being inaccurate when attempting to order loci over relatively short (<1 Mb) chromosomal regions. Due to the limited resolution of genetic maps that have been established through the use of these approaches, we have constructed a 2-Mb bacterial and P1-derived artificial chromosome (BAC-PAC) contig for the EGFR region and have applied markers positioned on its associated physical map to the analysis of 7p11.2 amplifications in a series of glioblastomas. Our data indicate that EGFR is the sole amplification target within the mapped region, although there are several additional 7p11.2 genes that can be coamplified and overexpressed with EGFR. Furthermore, these results are consistent with EGFR amplicons retaining the same organization as the native chromosome 7p11.2 region from which they are derived. PMID- 11916500 TI - Histone acetylation may suppress human glioma cell proliferation when p21 WAF/Cip1 and gelsolin are induced. AB - Histone deacetylase inhibitors that increase histone acetylation on transformed cells are being investigated as unique anticancer drugs. The aim of this investigation was to evaluate an antiproliferative activity of the histone deacetylase inhibitors sodium butyrate (NaBT) and trichostatin A on 5 glioma cell lines, T98G, A172, U-87 MG, U-118 MG, and U-373 MG, with the examination of the altered expressions in p21 and gelsolin genes. Treatment with 5-mM NaBT and 40 ng/ml trichostatin A for 48 h caused more than a 50% growth inhibition in 5 cell lines as measured by cell proliferation assays. An increase in histone acetylation was confirmed in each cell line. After treatment with 5 mM NaBT, T98G, A172, and U118 cells undergo apoptosis as indicated by DNA ladder formation. Treatment with NaBT and trichostatin A also decreased DNA synthesis as examined by the fluorescence-activated cell sorting analysis in T98G and U87 cells. In addition to the suppression of cell growth, the up regulation of p21 and gelsolin expression was observed after treatment with NaBT, especially in T98G cells. Maximum expression of p21 and gelsolin was observed within 24 h after treatment. Results from our in vitro studies indicate that the treatment of human glioma cells with one of the histone deacetylase inhibitors suppresses cell growth with decreasing DNA synthesis and stimulates apoptosis, and that associated molecular mechanisms responsible for these effects include increased histone acetylation as well as enhanced expression of p21 and gelsolin. PMID- 11916501 TI - Phase II study of irinotecan (CPT-11) in children with high-risk malignant brain tumors: the Duke experience. AB - A phase II study of irinotecan (CPT-11) was conducted at Duke University Medical Center, Durham, NC, to evaluate the activity of this agent in children with high risk malignant brain tumors. A total of 22 children were enrolled in this study, including 13 with histologically verified recurrent malignant brain tumors (glioblastoma multiforme [GBM] 4, anaplastic astrocytoma 1, ependymoma 5, and medulloblastoma/primitive neuroectodermal tumor 3), 5 with recurrent diffuse pontine glioma, and 4 with newly diagnosed GBM. All patients with recurrent tumor had prior chemotherapy and/or irradiation. Each course of CPT-11 consisted of 125 mg/m ( 2 ) per week given i.v. for 4 weeks followed by a 2-week rest period. Patients with recurrent tumors received therapy until disease progression or unacceptable toxicity. Patients with newly diagnosed tumors initially received 3 cycles of treatment to assess tumor response and then were allowed radiotherapy at physician's choice; patients who demonstrated a response to CPT-11 prior to radiotherapy were allowed to continue the drug after radiation until disease progression or unacceptable toxicity. A 25% to 50% dose reduction was made for grade III-IV toxicity. Responses were assessed after every course by gadolinium enhanced MRI of the brain and spine. Twenty-two patients received a median of 2 courses of CPT-11 (range, 1-16). Responses were seen in 4 of 9 patients with GBM or anaplastic astrocytoma (44%; 95% confidence interval, 11%-82%) (complete response in 2 patients with recurrent GBM lasting 9 months and 48+ months; partial response in one patient with a newly diagnosed midbrain GBM lasting 18 months prior to radiotherapy; and partial response lasting 11 months in 1 patient with recurrent anaplastic astrocytoma), 1 of 5 patients with recurrent ependymoma (partial response initially followed by stable disease lasting 11 months), and none of 5 patients with recurrent diffuse pontine glioma. Two of 3 patients with medulloblastoma/primitive neuroectodermal tumor had stable disease for 9 and 13 months. Toxicity was mainly myelosuppression, with 12 of 22 patients (50%) suffering grade II-IV neutropenia. Seven patients required dose reduction secondary to neutropenia. CPT-11, given in this schedule, appears to be active in children with malignant glioma, medulloblastoma, and ependymoma with acceptable toxicity. Ongoing studies will demonstrate if activity of CPT-11 can be enhanced when combined with alkylating agents, including carmustine and temozolomide. PMID- 11916502 TI - A phase II trial of thymidine and carboplatin for recurrent malignant glioma: a North American Brain Tumor Consortium Study. AB - This phase II study in recurrent high-grade glioma evaluated the response rate, toxicities, and time to treatment failure of high-dose carboplatin modulated by a 24-h infusion of thymidine (75 g/m(2)). The trial was based on preclinical data and a prior phase I study ( J. Clin. Oncol. 17, 2922-2931, 1999); a phase II recurrent high-grade glioma study was initiated in July of 1998. Thymidine was given over 24 h; carboplatin was given over 20 min at hour 20 of the thymidine infusion. The starting dose of carboplatin had a value of 7 for the area under the curve (AUC), with allowance for dose escalation of 1 AUC unit per cycle if grade 2 toxicity was observed. Treatment cycles were repeated every 4 weeks. Accrual as of September 1999 was 45 patients [4 were unevaluable]: 76% with glioblastoma multiforme (GBM), 20% with anaplastic oligodendroglioma, 2% with mixed type, and 2% with anaplastic astrocytoma. Most patients had prior chemotherapy (78%). As observed in the earlier phase I study (in which carboplatin pharmacokinetics were unaltered by thymidine or antiseizure medications), thymidine was myeloprotective, resulting in a minimal need for dose reduction for patients having a >2 grade toxicity (in only 4% of the courses of treatment). Of 101 total courses, the number of courses (at the AUCs) was 3 (5), 4 (6), 58 (7), 20 (8), 11 (9), and 5 (10). Grade 3 nonhematologic toxicities included headache (4%), altered consciousness (3%), fatigue (1%), and nausea (3%). Responses included 2 partial (1 oligodendroglioma, 1 GBM; 5%); 3 minor (1 anaplastic astrocytoma, 2 GBM; 7.3%); 6 stable disease (14.6%); and 30 progressive disease (73.2%). For GBM patients, median survival was 23 weeks (with a 95% confidence interval of 20 to 50 weeks), and progression-free survival was 8 weeks (with a 95% confidence interval of 7-16 weeks). These results in GBM were comparable to other phase II GBM trials and thus do not represent a therapeutic advance in the treatment of GBM. Taken collectively, however, results are consistent with continued investigation of thymidine in combination with chemotherapeutic agents for high-grade glioma and other malignant diseases. The significant myeloprotection afforded by thymidine may have particular relevance to polychemotherapeutic regimens. PMID- 11916503 TI - MDM2 overexpression is associated with short survival in adults with medulloblastoma. AB - In adult medulloblastoma, postoperative radiotherapy is significantly effective in prolonging time to recurrence and survival time; however, the response of individual cases to radiotherapy, that is the total survival, is different. Apoptosis is an important cellular response to radiation. It can be hypothesized that the individual radiosensitivity of medulloblastomas depends on the individual capability to undergo apoptosis. p53 protein is involved in the apoptotic response to ionizing radiation; loss of function of p53 can be the consequence not only of TP53 mutations, but also of amplification and/or overexpression of the MDM2 gene. We have analyzed cerebellar medulloblastomas from 51 adults (>16 years of age) for MDM2 gene amplification (by differential polymerase chain reaction assay), TP53 gene mutation (by polymerase chain reaction single-strand conformation polymorphism analysis of exons 5-8), and immunohistochemical expression of p53 (clone DO1) and MDM2 (clone IF2). The results have been evaluated in relation to age, tumor location, classic or desmoplastic type, MIB-1 labeling index, and total survival. No tumor had MDM2 amplification. Ten tumors had MDM2 positive tumor cells. One case had a mutated TP53 gene; 16/51 cases had intense p53 immunostaining. Only 2 MDM2 protein positive tumors were also p53-positive. Both subgroups of MDM2 - and p53-positive tumors had a significantly shorter postoperative survival. In conclusion, the overexpression of MDM2 protein and the accumulation of wild-type p53 are unrelated in adult medulloblastoma; they may result in a reduced apoptotic response after radiotherapy and contribute to a shortened survival. Also, MDM2 amplification and TP53 gene mutation are rare events in medulloblastomas of adults. PMID- 11916504 TI - Metastatic papillary craniopharyngioma: case study and study of tumor angiogenesis. AB - We report a case of suprasellar papillary craniopharyngioma metastatic to the temporoparietal region 2 years after its initial resection. The literature documents examples of craniopharyngioma recurrences along the surgical tract, as well as remote ipsi- and contralateral metastases via cerebrospinal fluid seeding. Ours is the second report of a craniopharyngioma of papillary type to exhibit metastatic behavior. The tumor spread opposite the side of craniotomy. Although a rare occurrence, it confirms the limited capacity of histologically benign craniopharyngiomas to undergo meningeal seeding, likely the result of surgical manipulation. Immunohistochemical demonstration of increased microvascular density and vascular endothelial growth factor expression, as well as a high vascular endothelial growth receptor (VEGFR2) signal by in situ hybridization, suggests that tumor vascularity facilitated angiogenesis and may have been involved in the establishment and growth of the metastatic deposit. PMID- 11916505 TI - Intractable vomiting from glioblastoma metastatic to the fourth ventricle: three case studies. AB - Dissemination of malignant glioma to the fourth ventricle with metastatic deposits and intractable vomiting is rare. Leptomeningeal extension of malignant glioma is an uncommon condition that has been reported in patients with end-stage disease and is usually unresponsive to any treatment modality. We describe 3 patients with progressing recurrent glioblastoma multiforme in whom leptomeningeal invasion manifested itself as intractable vomiting due to tumor metastases in the floor of the fourth ventricle. All patients received additional radiation therapy focused to the posterior fossa, with complete resolution of vomiting occurring within 10 days after irradiation. The remission of symptoms in these patients persisted until their death 3-4 months after the repeat radiation therapy. These reports indicate that additional focused radiation should be considered because of its significant therapeutic effect in alleviating intractable nausea and vomiting in patients with glioma metastasized to the posterior fossa. PMID- 11916506 TI - Consensus Conference on Brain Tumor Definition for registration. November 10, 2000. AB - The Consensus Conference on Brain Tumor Definition was facilitated by the Central Brain Tumor Registry of the United States and held on November 10, 2000, in Chicago, Illinois, to reach multidisciplinary agreement on a standard definition of brain tumors for collecting and comparing data in the U.S. The Brain Tumor Working Group, convened in 1998 to determine the status of brain tumor collection in the U.S., outlined 4 recommendations of which the first 2 guided the discussion for the Consensus Conference: (1) standardization of a definition of primary brain tumors that is based on site alone, rather than on site and behavior, and that can be used by surveillance organizations in collecting these tumors; and (2) development of a reporting scheme that can be used for comparing estimates of primary brain tumors across registries. Consensus was reached on the collection of all primary brain tumor histologies found and reported in the brain or CNS ICD-O site codes (C70.0-C72.9 and C75.1-C75.3), including those coded benign and uncertain as well as those coded malignant. In addition, a comprehensive listing of histologies occurring in the brain and CNS, based on the CBTRUS grouping scheme, was formulated to provide a template for reporting in accordance with the second recommendation of the Brain Tumor Working Group. With consensus achieved on the first 2 recommendations, the stage is set to move forward in estimating additional resources necessary for the collection of these tumors, including funding, training for cancer registrars, identifying quality control measures, and developing computerized edit checks, as outlined in the last 2 recommendations of the Brain Tumor Working Group. PMID- 11916507 TI - Non-linear dynamics models characterizing long-term virological data from AIDS clinical trials. AB - Human immunodeficiency virus (HIV) dynamics represent a complicated variant of the text-book case of non-linear dynamics: predator-prey interaction. The interaction can be described as naturally reproducing T-cells (prey) hunted and killed by virus (predator). Virus reproduce and increase in number as a consequence of successful predation; this is countered by the production of T cells and the reaction of the immune system. Multi-drug anti-HIV therapy attempts to alter the natural dynamics of the predator-prey interaction by decreasing the reproductive capability of the virus and hence predation. These dynamics are further complicated by varying compliance to treatment and insurgence of resistance to treatment. When following the temporal progression of viral load in plasma during therapy one observes a short-term (1-12 weeks) decrease in viral load. In the long-term (more than 12 weeks from the beginning of therapy) the reduction in viral load is either sustained, or it is followed by a rebound, oscillations and a new (generally lower than at the beginning of therapy) viral load level. Biomathematicians have investigated these dynamics by means of simulations. However the estimation of the parameters associated with the dynamics from real data has been mostly limited to the case of simplified, in particular linearized, models. Linearized model can only describe the short-term changes of viral load during therapy and can only predict (apparent) suppression. In this paper we put forward relatively simple models to characterize long-term virus dynamics which can incorporate different factors associated with resurgence: (Fl) the intrinsic non-linear HIV-1 dynamics, (F2) drug exposure and in particular compliance to treatment, and (F3) insurgence of resistant HIV-1 strains. The main goal is to obtain models which are mathematically identifiable given only measurements of viral load, while retaining the most crucial features of HIV dynamics. For the purpose of illustration we demonstrate an application of the models using real AIDS clinical trial data involving patients treated with a combination of anti-retroviral agents using a model which incorporates compliance data. PMID- 11916508 TI - Retention of antigen on follicular dendritic cells and B lymphocytes through complement-mediated multivalent ligand-receptor interactions: theory and application to HIV treatment. AB - In HIV-infected patients, large quantities of HIV are associated with follicular dendritic cells (FDCs) in lymphoid tissue. During antiretroviral therapy, most of this virus disappears after six months of treatment, suggesting that FDC associated virus has little influence on the eventual outcome of long-term therapy. However, a recent theoretical study using a stochastic model for the interaction of HIV with FDCs indicated that some virus may be retained on FDCs for years, where it can potentially reignite infection if treatment is interrupted. In that study, an approximate expression was used to estimate the time an individual virion remains on FDCs during therapy. Here, we determine the conditions under which this approximation is valid, and we develop expressions for the time a virion spends in any bound state and for the effect of rebinding on retention. We find that rebinding, which is influenced by diffusion, may play a major role in retention of HIV on FDCs. We also consider the possibility that HIV is retained on B cells during therapy, which like FDCs also interact with HIV. We find that virus associated with B cells is unlikely to persist during therapy. PMID- 11916509 TI - A transduction model of the Meissner corpuscle. AB - I propose a transduction model of the Meissner corpuscle that integrates ideas put forth by Freeman and Johnson and results obtained by Looft. The principal development in the present model is its specification that RA receptor potentials are updated as a linear function of stimulus velocity above baseline; the model thus readily accommodates non-sinusoidal input. It also incorporates modifications to Freeman and Johnson's model proposed by Slavik and Bell, namely a period of refractoriness lasting 1 ms followed by a period of hyperexcitability lasting 13.5 ms. The model is applied to various psychophysical and physiological situations: psychophysical threshold vs. frequency, RA afferent impulse rates vs. intensity, impulse regularity vs. frequency, phase retardation vs. frequency, and responses to non-repeating noise and to complex stimuli. Model output closely matches psychophysical and neurophysiological data. The proposed model thus reliably predicts RA afferent responses to arbitrary stimuli and may facilitate the development of theories relating psychophysical phenomena to their underlying neural representations. PMID- 11916511 TI - Statistical and algorithmic methods for fluctuation analysis with SALVADOR as an implementation. AB - This paper aims at removing certain long-standing impediments to more effective and widespread use of fluctuation analysis. The paper presents a method of constructing confidence intervals for mutation rates using data from fluctuation experiments. The method was inspired by a rediscovery of a little-known, not fully developed method of Lea and Coulson; substantial modifications have been made both to enhance computational efficiency and to widen the scope of the original method's applicability. A computer package named SALVADOR is presented that can be used for Monte Carlo simulation, for point and interval estimation of mutation rates, and for exploration of various hypotheses spawned by the directed mutation controversy. In addition to the maximum likelihood method, methods of considerable historical interest are also examined and included in SALVADOR to help the reader compare and assess some of the most popular methods for estimating mutation rates. PMID- 11916510 TI - Estimating an oncogenetic tree when false negatives and positives are present. AB - Human solid tumors are believed to be caused by a sequence of genetic abnormalities arising in the tumor cells. The understanding of these sequences is extremely important for improving cancer treatment. Models for the occurrence of the abnormalities include linear structure and a recently proposed tree-based structure. In this paper we extend the pure oncogenetic tree model by introducing false positive and false negative observations. We state conditions sufficient for the reconstruction of the generating tree. As an example we analyze a comparative genomic hybridization data set and show that addition of the error model significantly improves the ability of the model to describe the data. PMID- 11916513 TI - Erythroleukemia: a comparison between the previous FAB approach and the WHO classification. AB - Erythroleukemia is, within FAB classification, a proliferation of erythroblasts superior to 50% and of myeloblasts superior to 30%. The new WHO classification abolishes the frontier between RAEB-t with 20% and leukemia with 30% of blasts. AML6 variant is a new entity characterized by the proliferation of immature erythroblasts and the absence of non-erythroid blast cells. We analyzed 16 erythroleukemia, 5 RAEB-t and 2 AML6 variants to clarify their relationship. We suggest on survival, karyotype and cytologic characteristics that secondary erythroleukemia are the same entity as RAEB-t, confirming the WHO classification and that amongst de novo erythroleukemia, there is 'AML6 variant' with pure erythroid lineage proliferation. PMID- 11916512 TI - Mathematical modeling of myoglobin facilitated transport of oxygen in devices containing myoglobin-expressing cells. AB - Low pO(2) is perhaps the most significant factor in artificial pancreas failure. In these environments, not only is the beta cell production of insulin reduced, but the cell death rate is also significantly higher. Mathematical models are developed to test the feasibility of facilitated oxygen transport in enhancing O(2) flux to genetically engineered cells in a bioartificial device such as a pancreas. For this device, it is proposed that beta cells be genetically engineered to express myoglobin throughout the cell. In addition, the significance of including myoglobin throughout the alginate matrix present to provide immuno-protection for the transplanted cells is considered. The mathematical analysis predicts that myoglobin facilitated oxygen transport has the potential of increasing the oxygen concentration at the centre of a cluster of cells (islet) with an effective radius of 100 microm by 50%. These theoretical models for myoglobin facilitated oxygen transport with homogeneous Michaelis Menten consumption also indicate that including myoglobin in the alginate gel would beneficially improve the flux of oxygen to the transplanted cells. PMID- 11916514 TI - Minimal residual disease in Brazilian children with acute lymphoid leukemia: comparison of three detection methods by PCR. AB - The minimal residual disease (MRD) detection by the polymerase chain reaction (PCR) in children with acute lymphoblastic leukemia has been pointed to be an adverse prognostic factor. Detection methods based on this technique using clone specific primers are cumbersome and time consuming. The detection of monoclonal gene rearrangements of gamma T-cell receptors (TCRgamma) is a simpler although less sensitive method. In the present study, we analyzed the presence of MRD during four different phases of treatment (week 4; 3-6, 12-24 months, and end of treatment) in 34 Brazilian children with lymphoid leukemia by three detection methods based on the PCR technique: (1) using consensus primers for the detection of a clonal population for TCRgamma; (2) clone-specific primers for the junctional region of TCRgamma; and (3) a semi-nested reaction with an initial cycle with consensus primers followed by a second cycle with clone-specific primers. MRD presence was associated with a shorter event-free survival and was the major independent prognostic factor in most of the phases analyzed. The use of consensus primers for the detection of TCRgamma clonality, although less sensitive, proved to be a simpler, faster and less costly method whose positivity was associated with more than 90% relapse rates during all phases analyzed. PMID- 11916515 TI - Identification of two atypical PML-RAR(alpha) transcripts in two patients with acute promyelocytic leukemia. AB - We identified two patients with atypical PML-RAR(alpha) rearrangements, 53 and 13 base pairs longer than the typical bcr1 transcript. Sequence analysis revealed a new PML breakpoint at the end of exon 7a in patient 1, and a PML exon 6 breakpoint in patient 2, with an insertion of 35 nucleotides of RAR(alpha) intron 2. Patient 1 did not express RAR(alpha)-PML and patient 2 showed the RAR(alpha) PML transcript, which corresponded to the typical bcr1. These results emphasize on the relevance of the correct identification of atypical PML-RAR(alpha) rearrangements because of the potential implications in leukemogenesis, in the response to treatment, and for the correct monitoring of minimal residual disease. PMID- 11916516 TI - CD80 antigen expression as a predictor of ex vivo chemosensitivity in chronic lymphocytic leukemia. AB - We investigated the correlation between expression of 31 surface membrane antigens and chemosensitivity of peripheral blood mononuclear cells from 36 patients with CLL. The sensitivity of CLL cells to nine drugs (2' chlorodeoxyadenosine, cisplatin, chlorambucil, cyclosporin A, doxorubicin, fludarabine, prednisolone, verapamil and vincristine) and two types of irradiation (gamma and UV-irradiation) was determined from dose-response curves of 4-day cultures ex vivo. The results indicated that the CLL cases responding to purine analogs (2'-chlorodeoxyadenosine and fludarabine) can be identified according to CD80 expression: all resistant cases had low or negative CD80 expression. No other correlations were revealed. CD80 may be a surrogate chemosensitivity marker for purine analogs. PMID- 11916517 TI - Non-Hodgkin's lymphoma in patients 70 years of age or older: factors associated with survival. AB - The incidence of non-Hodgkin's lymphoma (NHL) has increased dramatically especially in persons over 60 years of age. We reviewed our experience in patients with NHL who were 70 years of age or older and seen between 1992 and 1998 in an effort to find any unique risk factors in this age group. There were 64 patients. Twenty-five had indolent disease and 38 had aggressive disease. The majority of patients with indolent lymphoma presented with nodal disease and were more likely to have bone marrow involvement. In contrast, 71% of patients with aggressive lymphoma presented with extra-nodal disease. Five of seven (71%) patients with indolent and 22 of 29 (76%) with aggressive NHL who received an Adriamycin containing regimen as first line therapy achieved a complete response. The International Prognostic Index (IPI) was a strong predictor of both survival (P<001) and response (P=007) for the group as a whole. The IPI accurately predicted the survival of patients with aggressive NHL (P=026). This was not the case with indolent lymphomas. This study suggests that elderly patients with NHL are more likely to have aggressive disease, a diffuse pathology and an extra nodal presentation. PMID- 11916518 TI - C/EBP-beta, C/EBP-delta, PU.1, AML1 genes: mutational analysis in 381 samples of hematopoietic and solid malignancies. AB - Mutations of transcription factors are associated with the pathogenesis of cancer. Genomic DNA from 381 cancers and cell lines representing leukemias, lymphomas and a variety of solid tumors were examined for mutations of genes coding for the C/EBP-beta, C/EBP-alpha, PU.1, and AML1 transcription factors using single strand conformation polymorphism (SSCP) and direct DNA sequencing. Mutation of C/EBP-beta (a chronic myelogenous leukemia cell line, Kcl22) and C/EBP-delta (a Burkitt's lymphoma cell line, Raji) were found. Interestingly, the sample with a C/EBP-beta alterations had two missense (P236L and G252A) and two silent mutations in a highly conserved region of the gene. The C/EBP-delta alteration in Raji was a missense mutation (A177G). These findings suggest that mutations of the C/EBP-beta, C/EBP-delta, PU.1, and AML1 rarely contribute to the development of hematopoietic or solid cancers. PMID- 11916519 TI - Mutations in human cancer: how close should we look? PMID- 11916520 TI - Long-term follow-up of intensive ara-C-based chemotherapy followed by bone marrow transplantation for adult acute lymphoblastic leukemia: impact of induction Ara-C dose and post-remission therapy. AB - We report single institution outcome of brief, intensive ara-C-based chemotherapy using bone marrow transplantation as primary intensification for untreated adult patients with acute lymphoblastic leukemia (ALL). Overall disease-free and overall survival were inferior to those reported with prolonged chemotherapy modeled on pediatric protocols. Survival and disease-free survival were superior for patients receiving allogeneic BMT compared with chemopurged autologous transplant or maintenance chemotherapy (patients ineligible for or declining BMT). In multivariate analysis, non-L2-FAB, higher ara-C dose, absence of CNS disease, non-Ph1+ karyotype, allogeneic BMT, T cell phenotype, and younger age were associated with improved disease-free survival. Autologous BMT was not superior to chemotherapy, and appears unlikely to provide adequate curative treatment for most adult ALL patients if not followed by maintenance. PMID- 11916521 TI - The role of high-dose cytarabine in induction therapy for adult ALL. PMID- 11916522 TI - Purging in autologous hematopoietic stem cell transplantation using adenosine triphosphate (ATP) and 4-hydroperoxycyclophosphamide (4-HC). AB - In this study, we show potent in vitro purging induced by adenosine triphosphate (ATP) for leukemic cells. The treatment of murine L1210 leukemic cells with 2mM of ATP in vitro for 3h was able to reduce the number of leukemic clonogenic cells by about one order of magnitude presumably by changing the permeability of the leukemic cell membrane. Furthermore, the incubation of L1210 cells with ATP (2mM) and low dose 4-hydroperoxycyclophosphamide (4-HC, 2 microg/ml) for 3h resulted in at least a four-log reduction of clonogenic L1210 cells. Only a slight degree of toxicity to pluripotent hematopoietic stem cells (CFU-S) was observed in both treatment protocols. To determine the efficacy of pharmacological purging by ATP, we designed a murine system to mimic the conditions expected in the clinical setting of autologous transplantation using simulated partial remission marrow (SPRM) which was prepared by mixing normal marrow cells and L1210 cells at a ratio of 9:1. After the SPRM cells were incubated in vitro at a concentration of 1 x 10(6)/ml with both ATP (2mM) and low dose 4-HC (2 microg/ml) for 3h, 5 x 10(4) of the cells were then injected into lethally irradiated 9 weeks male BDF1 mice. All the mice given untreated-SPRM died of leukemia by day 27, whereas none of the recipients transplanted treated-grafts had died by day 70, thus suggesting that the combination use of ATP and 4-HC may be a potentially effective way to purge leukemic cells in autologous stem cell transplantation. The mechanism of the selective killing of leukemic cells is assumed that 4-HC is effectively incorporated into leukemic cells by increasing the permeability of the cell membrane by ATP. Taken together, this simple and rapid procedure is able to purge leukemic cells from autologous bone marrow grafts. PMID- 11916523 TI - Classification of cutaneous mastocytosis: a modified consensus proposal. PMID- 11916525 TI - Real-time RT-PCR for the determination of topoisomerase II mRNA level in leukaemic cells. AB - We developed a real-time RT-PCR assay for the quantification of topoisomerase II (topo II) mRNA level. It was applied on peripheral leukaemic cells from 23 patients with acute myelogenous leukaemia (AML) and 23 with chronic lymphocytic leukaemia (CLL). RNA template dilutions from 0.25 to 25ng per reaction were used as standard curves for topo IIalpha, beta and the internal control 18S rRNA. About 57% (26/46) and 26% (12/46) of the specimens had detectable topo IIbeta and alpha mRNA, respectively. The correlation between these two factors was rho=0.7 and P=0.0001. No relationship between topo IIalpha or beta mRNA level and response to chemotherapy was found in AML patients (n=19 assessable for response). Our method is rapid and convenient for quantification of topo IIalpha and beta mRNA levels, and could be suitable for investigation in a larger population. PMID- 11916526 TI - Valproic acid induces apoptosis in human leukemia cells by stimulating both caspase-dependent and -independent apoptotic signaling pathways. AB - We investigated the effects of valproic acid (VPA) on the growth and survival of human leukemia cell lines. VPA induced cell death in all of the nine cell lines tested in a dose dependent manner. VPA-treatment induced apoptotic changes in MV411 cells including DNA fragmentation, phosphatidylserine externalization, cytochrome c release from mitochondria, and activation of caspases-3, -8, and -9. A caspase inhibitor, zVAD-FMK, inhibited the DNA fragmentation induced by VPA but not cell death. These findings suggest that VPA exerts an anti-leukemic effect by both caspase-dependent and -independent apoptotic signaling pathways. PMID- 11916527 TI - Comparison of DR5 and Fas expression levels relative to the chemosensitivity of acute lymphoblastic leukemia cell lines. AB - The relationship between p53 gene status and the expression of DR5 and Fas was evaluated as a function of sensitivity of 11 acute lymphoblastic leukemia cell lines to adriamycin, etoposide, vincristine, methotrexate and dexamethasone. There was up to a 37-fold increase in expression of DR5 following treatment with ADR or VP-16 only in cells with wt p53. A direct correlation was observed between enhanced DR5 expression and sensitivity to ADR and VP-16. There was no induction of DR5 following treatment with VCR, MTX or DEX. There was up to a 51-fold increase in the median level of expression of Fas following treatment with ADR and VP-16, and unlike DR5 this occurred in cells with either wild-type or mutant p53. Nevertheless, a direct correlation was observed between Fas expression and drug-sensitivity. Conversely, there was only a two-fold increase in expression of Fas after exposure to VCR, MTX and DEX. These findings suggest that DR5 mediates sensitivity to ADR and VP-16 in a p53-dependent manner, whereas, Fas appears to mediate sensitivity to these two drugs independent of p53 status. DR5 and Fas do not appear to play a major role as determinants of chemosensitivity to VCR, MTX and DEX. PMID- 11916528 TI - Expression of Fc(epsilon)-receptors by human acute myelogenous leukemia (AML) blasts: studies of high- and low- (CD23) affinity receptor expression and the effects of IgE-mediated receptor ligation on functional AML blast characteristics. AB - Acute myelogenous leukemia (AML) blasts derived from 20 patients were examined for expression of high- (Fc(epsilon)RI) and low-affinity (Fc(epsilon)RII, CD23) IgE Fc(epsilon)-receptors. Fc(epsilon)RI expression was not detected for any patient. In contrast, expression of CD23 (at least 15% of the blasts stained positive) was detected for 6 out of the 20 patients. Acute lymphoblastic leukemia (ALL) blasts derived from 12 patients did not express CD23 (<1% positive cells for all patients). The functional effects of Fc(epsilon)R-receptor ligation were also examined for 20 patients, including the five patients with highest CD23 expression (30-55% positive cells) and five patients with verified low CD23 expression (0.010 ng/ml, was measured in only six samples from 3 patients. After completion of chemotherapy, 7 out of the 38 patients had left ventricular dysfunction (LV dysfunction). Only 1 of these 7 patients had an elevated cTnT level. 2 other patients with elevated cTnT levels did not develop LV dysfunction until 2 and 7 months after the cTnT measurement. Our data show that the measurement of cTnT within 24 h after administration of chemotherapy does not have a high sensitivity for the identification of patients with subsequent subclinical cardiotoxicity. PMID- 11916552 TI - A computer program for period analysis of cancer patient survival. AB - Monitoring of long-term survival rates, which is now routinely performed by many cancer registries throughout the world, should be as up-to-date as possible. A few years ago, a new method of survival analysis, denoted period analysis, has been proposed which provides more up-to-date estimates of long-term survival rates than traditional survival analysis by exclusively reflecting the survival experience of patients within a recent calendar period. However, application of this method has so far been hindered by the lack of pertinent computer programs. In this paper, we present a simple and easy-to-use computer program (SAS macro) that enables one to carry out period analysis (as well as conventional analysis) of both absolute and relative survival rates with the type of data commonly available in population-based cancer registries. We illustrate application of the program with examples from the nationwide Finnish Cancer Registry. PMID- 11916554 TI - Transition in cancer patterns among Turks residing in Germany. AB - Cancer mortality among the 2.1 million Turks residing in Germany is assumed to change from a pattern typical for a developing country towards one of an industrialised country. To test this hypothesis, we compared age-standardised cancer mortality rates among Turkish residents and (West) Germans using death registration data. In addition, we assessed proportional cancer incidence ratios among Turkish cases (n=144) in a German population-based cancer registry. All cancer mortality 1992-1997 (per 100000) was 34.8 (n=4192) among Turkish men (Germans: 72.3) and 21.5 (n=1862) among Turkish women (Germans: 52.4). Over time, gastric and lung cancer mortality increased among Turkish men, as did breast cancer mortality among Turkish women. The proportional cancer incidence (PCIR) for stomach cancer among men was 2.9 (95% Confidence Interval (CI): 1.7-4.8), and that for breast cancer among women was 0.7 (95% CI: 0.4-1.1). Turks had an increased proportional incidence ratio for non-Hodgkin's lymphoma. Our findings partly support a transition of cancer patterns among Turks in Germany. PMID- 11916553 TI - Contacts with animals and humans as risk factors for adult brain tumours. An international case-control study. AB - While numerous studies have addressed the possible role of farming and related exposures as risk factors for brain tumours in adults, few of them have examined the potential effect of exposure to farm animals or pets. In an international multicentre case-control study, we investigated whether residence on a farm, contact with animals, or working in occupations with a high degree of potential contact with animals or humans were associated with brain tumours. Using a common questionnaire, 1177 cases of glioma, 330 with meningioma and 2478 controls from eight centres were interviewed about the exposures and, in particular, about their contacts with nine species of animals: dairy cattle, beef cattle, pigs, horses, sheep, goats, poultry, dogs and cats. Living or working on a farm was not a risk factor, for either glioma or meningioma. Except in some centres, there was no relationship between having contacts with farm animals or pets and the risk of brain tumour, for either type of tumour or either sex. In relation to seven industrial groups involving frequent human and/or animal contacts, no association was apparent for either glioma or meningioma. In relation to 25 occupational groups with potential frequent contact with humans and/or animals, for glioma there was a reduced risk for biological technicians (Odds Ratio (OR)/=0, P=0.01), and general farm workers (OR=0.66, 95% Confidence Interval (CI): 0.5-0.9). For meningioma, there was an increased risk for cooks (OR=2.0; CI: 1.2-3.4). With some exceptions, these results indicate no association between either the type of brain tumour and contacts with animals, or with occupations that include a high level of contact with animals or a high level of contact with humans. PMID- 11916555 TI - Overexpression of pyrimidine nucleoside phosphorylase enhances the sensitivity to 5'-deoxy-5-fluorouridine in tumour cells in vitro and in vivo. AB - 5-Fluorouracil (5-FU) and 5'-deoxy-5-fluorouridine (5'-DFUR), a prodrug of 5-FU, are representative of the chemotherapeutic agents for colorectal adenocarcinomas. Pyrimidine nucleoside phosphorylase (PyNPase) catalyses the conversion of 5'-DFUR to 5-FU, the activated form. Murine adenocarcinoma CT26 cells were transfected with human PyNPase cDNA. The engineered transfectants producing PyNPase augmented the response to 5'-DFUR in vitro and in vivo. Animals were administered by means of intraperitoneal (i.p.) injection, and not orally, in order to obtain a better efficiency of absorption. The tumours of the transfected cells nearly all disappeared, even following treatment with quite a small amount of the anticancer agent. The animals injected with the tranfected cells were protected against subsequent challenge with the parental tumour cell line. These findings demonstrate that PyNPase gene transfection increases the sensitivity to 5'-DFUR, and thereby decreases the toxicity of the agent. PMID- 11916556 TI - Radiobiological characteristics of solid tumours depending on the p53 status of the tumour cells, with emphasis on the response of intratumour quiescent cells. AB - Human head and neck squamous cell carcinoma cells transfected with mutant TP53 (SAS/mTP53) or with a neo vector as a control (SAS/neo) were inoculated subcutaneously (s.c.) into both hind legs of Balb/cA nude mice. Mice bearing tumours received 5-bromo-2'-deoxyuridine (BrdU) continuously to label all proliferating (P) cells in the tumours. The mice then received gamma-ray irradiation. Another group of mice received a series of test doses of gamma-rays while alive or after tumour clamping to obtain hypoxic fractions (HFs) in the tumours. Right after irradiation, the tumour cells were isolated and incubated with a cytokinesis blocker. The micronucleus (MN) frequency in the cells without BrdU labelling (=quiescent (Q) cells) was determined using immunofluorescence staining for BrdU. Meanwhile, 6 h after irradiation, tumour cell suspensions obtained in the same manner were used for determining the frequency of apoptosis in the Q cells. The MN frequency and apoptosis frequency in total (P+Q) tumour cells were determined from the tumours that were not pretreated with BrdU. In total cell populations, SAS/mTP53 cells were more radioresistant than SAS/neo cells in clonogenic survival. Q tumour cells exhibited a significantly lower apoptosis and MN frequency, probably due to their much larger HF, than total cells. In both total and Q cell fractions, SAS/mTP53 cells were less susceptible to apoptosis and more susceptible to micronucleation than SAS/neo cells. Obviously, TP53 status had the potential to influence the radiosensitivity of not only the total cells, but also the Q cells. However, irrespective of the TP53 status, significant differences in radiosensitivity between total and Q tumour cells were consistently observed. From the viewpoint of tumour control as a whole, including intratumour Q tumour cell control, a treatment modality for enhancing the Q cell response has to be considered. PMID- 11916557 TI - High frequency of LOH, MSI and abnormal expression of FHIT in gastric cancer. AB - The FHIT gene is a putative tumour suppressor gene. In this study, we analysed a set of 50 gastric tumours for alterations of FHIT, and found 38 of 45 tumours (84%) exhibiting loss of heterozygosity (LOH) within the FHIT gene. We used both nested Reverse Transcriptase Polymerase Chain Reaction (RT-PCR) and single step RT-PCR to analyse the FHIT transcripts and found 34 of 39 (87%) tumours and seven of the 11 (64%) corresponding non-cancerous tissues showed low or aberrant expression of FHIT mRNA and the appearance of the aberrant FHIT transcripts depended on the conditions of the RT-PCR. In these aberrant transcripts, frequent deletions and/or insertions were detected by direct sequencing. All breakpoints for deletions and insertions were at splicing sites. All insertions came from the adjacent introns, whose appearance was completely in accordance with the 'GU-AG' rule for pre-mRNA splicing. It may be suggested that an alternative splicing mechanism functions in the formation of these aberrant transcripts. The fragile nature of FRA3B within the FHIT gene could be responsible for the formation of the aberrant mRNA. Negative or reduced Fhit expression was detected in 39 of 50 tumours (78%). Moreover, an association was found between abnormal Fhit expression and positive node status (P=0.012). Thirteen of 48 tumours (27%) displayed microsatellite instability (MSI), among which 10 tumours also showed MSI within the FHIT gene. Furthermore, we detected an association between MSI and negative node status (P=0.02). We conclude that the abnormalities of FHIT, presumably associated with the unstable nature of FRA3B within the FHIT gene, are involved in the carcinogenesis of gastric cancer, and lack of mismatch repair (MMR) could possibly promote its alteration in a subset of gastric tumours. PMID- 11916559 TI - Is there a bioethicist in your company? Should there be? PMID- 11916560 TI - Receptor sponge soaks up cancer cells. PMID- 11916561 TI - Computer chip that could restore sight. PMID- 11916562 TI - Gene therapy to manipulate the immune system? PMID- 11916565 TI - Sir Chris Evans divulges his experiences as an entrepreneur. Interview by Rebecca N. Lawrence. PMID- 11916566 TI - The NR2B subunit of glutamate receptors as a potential target for relieving chronic pain: prospects and concerns. PMID- 11916567 TI - Three-dimensional information is not essential for drug discovery. PMID- 11916568 TI - Human tissue in target identification and drug discovery. PMID- 11916571 TI - Toxicoproteomics -- a new preclinical tool. AB - The publication of the human genome has presented the scientific community with an unprecedented amount of genetic information with the potential to revolutionize the drug discovery process. This information could be used to identify novel drug targets and disease markers or could aid in the development of personalized medicines. The realization that genetic changes must ultimately influence protein function has pushed the field of proteomics further into the limelight. In this review the applications of proteomics to the field of toxicology will be discussed. It is anticipated that, in the future, toxicologists will apply a range of genomic and proteomic techniques to address issues in toxicity. PMID- 11916572 TI - Validation of targets and drug candidates in an engineered three-dimensional cardiac tissue model. AB - High-throughput target discovery confronts the biopharmaceutical industry with a plethora of target candidates. The validation of these candidates in disease specific animal models often lacks the required throughput. Here, we discuss perspectives and limitations of a novel engineered three-dimensional cardiac tissue, which enables the influence of gene and drug intervention to be monitored on a cellular and molecular level under physiological conditions in sufficient throughput. The model is an extremely helpful filter to prioritize multiple development candidates before moving a project into large animal models with higher predictivity. PMID- 11916573 TI - Gene delivery using temperature-responsive polymeric carriers. AB - Synthetic temperature-responsive polymers can be applied as gene carrier systems. For successful gene therapies, efficient and safe vectors are essential because they deliver genes to target cells and aid gene expression of therapeutic peptides. Vector systems that can control gene expression are favorable, especially for genes whose therapeutic effects are considerably dependent on quantity, site, duration and timing of their expression. Strategy and clinical feasibility of the temperature-responsive vector system are discussed with an example of gene expression enhancement by temperature stimuli. PMID- 11916578 TI - Myocardial integrated backscatter: the renaissance of an old parameter? PMID- 11916579 TI - A new millennium without blind pericardiocentesis? PMID- 11916580 TI - Seeing the invisible: a short history of cardiac ultrasound. PMID- 11916581 TI - Can stress echocardiography compete with perfusion scintigraphy in the detection of coronary artery disease and cardiac risk assessment? AB - AIMS: The aim of this review was to define the place of stress echocardiography in the context of perfusion scintigraphy for the detection of coronary artery disease (CAD) and the assessment of cardiac risk. Stress echocardiography has the benefits of widespread availability, relatively low cost, portability, absence of radiation, and the determination of the ischaemic threshold. However, the echocardiographic windows are variable, sometimes with poor echogenicity, and interpretation is subjective and requires an adequate learning period. METHODS AND RESULTS: Diagnostic and prognostic comparisons were focused on studies comparing stress (exercise, dobutamine, adenosine or dipyridamole) echocardiography and perfusion scintigraphy in the same patients. These direct diagnostic comparisons (22 studies for a total of 1380 patients) show that stress echocardiography may be somewhat less sensitive in detecting and localizing mild CAD (in particular when vasodilators are used), but is more specific than perfusion scintigraphy. The direct prognostic comparisons (five studies for a total of 805 patients) show that stress echocardiography and perfusion scintigraphy have comparable prognostic value. CONCLUSIONS: At this moment, stress echocardiography already seems very competitive with perfusion scintigraphy. In the near future, improvement in endocardial border detection and quantitation of wall motion analysis are expected to improve the value of stress echocardiography still further. PMID- 11916582 TI - Cost-effectiveness of stress echocardiography for assessment of coronary artery disease: what we know and what we need to know. AB - AIMS: This review explores the balance between the incremental data supplied by stress echocardiography and its cost. This technique is now established as an accurate tool for the diagnosis of coronary artery disease and myocardial viability, but the current medical-economic environment mandates careful consideration of the impact of the results on patient outcomes. METHODS AND RESULTS: The tools for assessment of cost and efficiency are reviewed. Two considerations are explored for controlling costs; avoidance of testing in those who will derive limited incremental data, and appropriate selection of stress echocardiography rather than other stress techniques. The balance between diagnostic accuracy and cost is explored in observational studies and computer models. Finally, the prognostic implications of testing are evaluated and data from the stress testing literature are explored to show that more effective patient selection for interventions may justify greater expenditure on testing. CONCLUSIONS: The appropriate selection of patients for testing has a significant evidence base. The cost of identifying coronary disease has been examined in a number of studies and facilitates the selection between testing modalities. Prognostic data are increasingly available, and further work is needed to combine this with cost analysis in order to show that stress echocardiography, like other stress modalities, may be used to guide therapy and thereby improve cost effective outcomes. PMID- 11916583 TI - Pulsed doppler myocardial imaging. A new approach to regional longitudinal function? PMID- 11916584 TI - The search for perfection is the enemy of progress. PMID- 11916585 TI - Double orifice mitral valve with flail leaflet: a transoesophageal echocardiographic examination. AB - We describe a case of double orifice mitral valve and a flail leaflet in a 54 year-old man. This rare congenital abnormality was disclosed through the discovery of a murmur. The transoesophageal approach clearly showed two approximately equal orifices with multiple papillary muscles. Colour Doppler echocardiography showed a moderate mitral regurgitation due to the prolapse of the posterior leaflet of the anteromedial orifice. No other abnormality was associated. Transoesophageal echocardiography is useful to analyse as well the anatomy as the functional aspect of the mitral apparatus in this particular case of congenital disease. PMID- 11916586 TI - Echocardiographic features of left atrial dissection. AB - AIMS: To study left atrial dissection, a rare complication of mitral valve replacement. METHODS AND RESULTS: From our hospital database of 5497 transoesophageal echocardiograms, we analysed 524 echocardiograms performed on 478 patients with mitral valve prosthesis. We found four patients (0.84%) with left atrial dissection diagnosed by transoesophageal echocardiography that visualized the left atrial dissection: in three patients the diagnosis was confirmed intraoperatively. Three patients had previously had replacements of the mitral valve. Left atrial dissection was a severe complication: one patient died and the two patients successfully operated on had paraprosthetic regurgitation. CONCLUSION: Transoesophageal echocardiography is the first choice for diagnosis of left atrial dissection, a rare complication of mitral valve replacement with an acute/subacute clinical course. Previous mitral valve replacement seems to be the main risk factor to develop left atrial dissection. PMID- 11916587 TI - Infected thrombi in the right atrium related to central venous catheters: an ongoing challenge. PMID- 11916588 TI - Quantifying cardiac flow with freehand 3D scan. PMID- 11916590 TI - Application of stress echocardiography to the evaluation of non-coronary heart disease. AB - Stress echocardiography is accepted as a routine test in patients with coronary artery disease for the identification of myocardial ischaemia, the diagnosis of viable myocardium and assessment of prognosis. This review addresses the application of the test to non-coronary heart diseases. Some caution should be applied to the use of stress echocardiography for the detection of ischaemia in patients with valvular heart disease. However, examination of haemodynamics and left ventricular function during exercise is worthwhile in the assessment of symptomatic patients with apparently mild or moderate valvular disease, as well as being useful for the detection of subclinical left ventricular dysfunction. In the future, this tool may be more widely applied to the identification of other 'subclinical' conditions, including pulmonary hypertension, anthracycline cardiotoxicity and valvular heart diseases, although the benefit of early diagnosis of these conditions remains to be established. PMID- 11916589 TI - Regional strain and strain rate measurements by cardiac ultrasound: principles, implementation and limitations. AB - The non-invasive quantification of regional myocardial function is an important goal in clinical cardiology. Myocardial thickening/thinning indices is one method of attempting to define regional myocardial function. A new ultrasonic method of quantifying regional deformation has been introduced based on the principles of 'strain' and 'strain rate' imaging. These new imaging modes introduce concepts derived from mechanical engineering which most echocardiographers are not familiar with. In order to maximally exploit these new techniques, an understanding of what they measure is indispensable. This paper will define each of these modalities in terms of physical principles and will give an introduction to the principles of data acquisition and processing required to implement ultrasonic strain and strain rate imaging. In addition, the current status of development of the technique and its limitations will be discussed, together with examples of potential clinical applications. PMID- 11916591 TI - Dobutamine stress echocardiography identifies anthracycline cardiotoxicity. AB - BACKGROUND: Anthracyclines are effective anti-cancer agents, but their therapeutic value is limited by their myocardial toxicity. We assessed the physiological responses of stress echocardiography at low doses of dobutamine (DSE) in patients treated with anthracycline. METHODS AND RESULTS: In a prospective study, 28 patients were studied before and 1 month after the end of chemotherapy. All patients had normal ejection fraction (EF) at rest before therapy and the mean dose of anthracycline was 212+/-15 mg/m(2). Echocardiographic Doppler studies were performed before and during dobutamine infusion (5 and 10 microg/kg per min). Rest echocardiography demonstrated a significant decrease of EF between the two examinations in ejection fraction (67+/-3% vs. 61+/-3%, P<0.001). The increase of the EF during dobutamine infusion was higher after chemotherapy compared to the initial examination (19+/-3% vs. 29+/-3%: P<.05). No difference in EF was observed at 10 microg/kg per min between before and after chemotherapy. In contrast, at rest no difference in diastolic parameters was observed between the two examinations. Moreover, a significant decrease of the peak E and the ratio E/A was observed during dobutamine infusion after chemotherapy (93+/-4 cm/s vs. 79+/-5 cm/s and 1.3+/-0.1 vs. 1.0+/-0.1, respectively;P<0.05). CONCLUSION: Stress echocardiography may prove to be a sensitive technique and useful non-invasive approach for evaluating subclinical anthracycline cardiotoxicity. PMID- 11916592 TI - The effect of pacing-induced heart rate variation on longitudinal and circumferential regional myocardial function after acute beta-blockade--a cardiac ultrasound study. AB - AIMS: To evaluate the effect of acute beta-blockade in combination with differing heart rates on longitudinal and circumferential regional myocardial function using Doppler myocardial imaging and two-dimensional-echocardiography. METHODS AND RESULTS: In seven pigs the following echocardiographic indices were measured at baseline, after beta-blockade both without and with atrial pacing: wall thickening fraction, fractional shortening, myocardial peak systolic velocity, transmyocardial velocity gradient and systolic velocity time integral of the posterolateral wall in short-axis view; mitral valve plane excursion, myocardial peak systolic velocity and systolic velocity time integral of the posterolateral wall in an apical five-chamber view. Peak systolic velocities and velocity gradients decreased significantly following acute beta-blockade but no further decay occurred at high heart rate due to pacing. The velocity time integrals and mitral valve plane excursion showed a tendency to decrease following beta blockade but only after pacing were they significantly reduced. The wall thickening fraction and fractional shortening showed a significant reduction after beta-blockade but no further decay after pacing. CONCLUSION: Changes in systolic velocities and velocity gradients were independent of heart rate reduction under high dosage beta-blockade, whereas wall thickening fraction, mitral valve plane excursion and velocity time integrals changed due to pacing. PMID- 11916593 TI - Changes in left ventricular filling pattern during dobutamine stress Doppler echocardiography. AB - AIMS: To study the left ventricular filling pattern during stress Doppler echocardiography and its utility in the detection of myocardial ischaemia. METHODS AND RESULTS: Fifty-seven consecutive patients underwent dobutamine stress echocardiography. The left ventricular filling pattern (E-wave velocity; A-wave velocity; E/A ratio; E-wave deceleration time) was analysed at baseline and at maximum heart rate reached. The percentage increase in these parameters from baseline to peak heart rate was also determined. Myocardial ischaemia (regional contractility worsening) was induced in 19 cases (ischaemic group) but not in 38 cases (non-ischaemic group). There were no differences between both groups at baseline. E-wave deceleration time decreased in the non-ischaemic group (197+/-63 vs. 167+/-65 ms, P=0.01) and increased in the ischaemic group (203+/-42 vs. 315+/ 135 ms, P<0.0001). A percentage increase in E-wave deceleration time of >30% showed a positive predictive value of 93% and a negative predictive value of 86% for detecting ischaemia, and in the multivariate analysis it was the only Doppler parameter (P<0.0001) that predicted the induction of ischaemia. CONCLUSION: We conclude that myocardial ischaemia provokes an increase in E-wave deceleration time. Analysis of left ventricular filling could help in the identification of those cases which are positive for ischaemia. PMID- 11916594 TI - Measurement of volumetric mitral and aortic blood flow based on a new freehand three-dimensional colour flow imaging method. An in vivo validation. AB - AIMS: To validate a new three-dimensional (3D) colour flow method used to calculate cardiac output (CO) in aortic and mitral blood flow. METHODS: The transducer was freely tilted transthoracically using a magnetic locating device recording its spatial position. Raw digital ultrasound data were recorded in healthy subjects during 10-20 heartbeats at a high frame rate ranging from 41 to 66 frames/s and analysed off-line with no loss in temporal resolution. Blood flow velocities aligned with the ultrasound beam were integrated across a moving spherical surface to calculate volumetric flow. RESULTS: The range of agreement between the 3D mitral and 3D aortic method was 0.04+/-1.32 l/min (mean+/-2 standard deviations). The range of agreement between 3D aortic flow and the two dimensional (2D) pulsed wave Doppler method (2DPW) in the left ventricular outflow tract (LVOT) was 0.7+/-1.7 l/min, while the range of agreement between 3D mitral flow and the 2DPW method was 0.88+/-1.64 l/min. CONCLUSION: The 3D methods agreed well. The 3D volumetric flow overestimated the 2DPW method, as expected, and the range of agreement was wide. The common pitfalls in pulsed wave ultrasound methods to calculate CO were avoided, as the 3D method was angle independent, no assumptions about the velocity profile were made, and a moving sample surface was applied. The acquisition of data was fast and easy and high temporal resolution was achieved. PMID- 11916595 TI - Rapid quantification of left ventricular function and mass using transoesophageal three-dimensional echocardiography: validation of a method that uses long-axis cutplanes. AB - AIMS: Despite its proven superiority compared to conventional echocardiographic techniques, three-dimensional (3D) echocardiography has not gained widespread acceptance in clinical medicine for the quantification of left ventricular volumes, function and mass. This is mainly due to the large, time-consuming process of data analysis. We sought to validate a new method that enables the accurate quantification of the left ventricle in a clinically acceptable short period of time. METHODS AND RESULTS: Left ventricular volumes, ejection fraction and mass were determined in 44 patients using 3D echocardiography. The 3D echocardiographic data sets were analysed: (i) using the conventional 'summation of slices' algorithm (slice thickness 5 and 10mm), which is based on the analysis of the 3D reconstructed left ventricle in short-axis cross-sections; and (ii) using the new method which is based on the analysis of the 3D reconstructed left ventricle in long-axis images. In each patient measurements were repeated using 3, 6, 7, 8, 9, 12 and 15 long-axis images. For all volumetric measurements there was a continuous reduction of measurement variability using increasing numbers of long-axis images. The use of more than nine long-axis images for volumes, and eight long-axis images for ejection fraction and mass, did not result in a further reduction of variability. The analysis time for volumes and masses averaged less than 5 min for the long-axis method using nine component images, compared to 20-43 min for the short-axis method. CONCLUSION: 3D echocardiography combined with a novel method based on the analysis of long-axis cross-section allows accurate quantification of left ventricular volumes, function and mass in a clinically acceptable short period of time. In the future, the combination of a real-time 3D echocardiographic acquisition technique with this analysis method should have important implications for the introduction of 3D echocardiography in clinical practice. PMID- 11916596 TI - Right atrial mass related to indwelling central venous catheters in patients undergoing dialysis. AB - Central venous catheters are used for the administration of fluids, drugs, blood products, total parenteral nutrition and for haemodynamic measurements. In patients with renal failure planned for dialysis, indwelling central venous catheters are used prior to forming an arterio-venous shunt. Infected right atrial thrombus is a rare phenomenon in adults and particularly unusual in patients undergoing dialysis. We describe two patients, undergoing dialysis for short periods, with indwelling central venous catheters and a right atrial mass, suspected for infected right atrial thrombus, detected by transoesophageal echocardiography. PMID- 11916597 TI - Alcohol ablation for mid-ventricular obstruction: straying from the path of righteousness? PMID- 11916598 TI - Left ventricular volume analysis during dobutamine stress echocardiography: a trend towards quantification. PMID- 11916599 TI - The balance between speed and efficacy in stress echocardiography: is earlier use of atropine the answer? PMID- 11916600 TI - Contrast echocardiography for myocardial perfusion imaging using intravenous agents: progress and promises. AB - AIMS: This article is a convenient overview to assist the interested echocardiographist towards acquiring his own experience in the field of myocardial perfusion imaging using intravenous contrast agents. This goal is now pursued in many centres, since contrast echo holds the advantages of cardiac ultrasound (non-invasiveness, high spatial and temporal resolution, wide availability, use of non-ionizing radiation), and because a variety of transpulmonary agents-together with a spectrum of imaging modalities-are becoming available. METHODS AND RESULTS: Many technical considerations need to be addressed for optimal myocardial perfusion imaging: characteristics of the contrast medium (air-filled or perfluorocarbon filled and/or encapsulated agents), modality of administration (bolus injection or continuous infusion) and interaction between microbubbles and ultrasound (dependency on power output). Moreover, intermittent harmonic imaging, intermittent harmonic power Doppler, pulse inversion and amplitude modulation imaging have all been developed to enhance microbubble detection over myocardial tissue. These new acquisition modalities also yield specific artifacts impacting on myocardial perfusion assessment. Finally, acute myocardial infarction and chronic ischaemic heart disease (at baseline and during stress) are the most studied clinical models for perfusion imaging with contrast echo, and are reviewed in this article. CONCLUSION: Perfusion imaging with intravenous contrast agents has never been as close to widespread clinical use as it is today, but many methodological issues remain unsettled before the wish of the contrast echocardiographist comes true: that is, a cheap, user-friendly and widely available technology that would disclose new information in echocardiography. PMID- 11916601 TI - Direct computation of multiple 3D flow convergence isovelocity surfaces from digital 3D reconstruction of colour Doppler data of the flow convergence region: an in vitro study with differently shaped orifices. AB - AIMS: Evaluation of the accuracy of direct computation of multiple three dimensional (3D) flow convergence (FC) isovelocities by using digital reconstruction of colour Doppler data of the flow convergence region. METHODS AND RESULTS: We used a conventional ultrasound system (ATL HDI 3000) connected to a computer workstation via Ethernet link. The digital 3D datasets were directly transferred to a Silicon Graphic Imaging Octane workstation for later measurement. We generated nine pulsatile flows (20-60 ml/beat), with peak flow rates (67-180 ml/s), through three orifices (circular, rectangular and triangular, S=0.24 cm(2)). The 3D reconstructions of FC surface areas from multi threshold velocities, including aliasing velocities were analysed to quantify the peak flow rate. For all orifices, linear regression demonstrated excellent correlation between the 3D calculated and electromagnetic flow meter recorded data. While there was a high correlation for 3D computation of flow rate from the single best Nyquist of 24 cm/s (r=0.97-0.98, SEE=7 .75-12.58 ml/s), the ability to average three threshold velocities (15, 18 and 24 cm/s) yielded an improved correlation (r=0.98-0.99, SEE=5.70-7.73 ml/s). CONCLUSIONS: Direct computation of multiple 3D FC isovelocities from digital reconstruction of colour Doppler data of the FC region provides the potential to accurately quantify the complex asymmetric spatial flow events at any selected velocity. PMID- 11916602 TI - Velocity profiles in mitral blood flow based on three-dimensional freehand colour flow imaging acquired at high frame rate. AB - AIMS: To describe the mitral blood flow velocity distribution, we applied a freehand dynamic three-dimensional (3D) colour flow method using a moving sample surface that followed the mitral apparatus during diastole. METHODS: Nineteen healthy volunteers were studied. The ultrasound data were captured from 10-20 heartbeats at high frame rate (mean 46 frames/s) while freely tilting the transducer in an apical position. A magnetic position sensor system recorded the spatial position and orientation of the probe. Blood flow velocities were integrated across a spherical surface. In volumetric blood flow measurements this would yield angle independence of the Doppler beam. Raw digital data were analysed off-line with no loss of temporal resolution. RESULTS: The ratio of the maximum velocity time integral (VTI) to the mean VTI was mean 1.3 (range 1.1 1.6). At the time of peak flow the ratio of the maximum to the mean velocity was mean 1.5 (range 1.2-2.6). CONCLUSION: The blood flow velocity profile was non uniform. By using a single sample volume in Doppler measurements of the maximum VTI errors ranging from 10 to 60% may be introduced in calculations of stroke volumes. PMID- 11916603 TI - Prospective study of early atropine use in dobutamine stress echocardiography. AB - AIMS: Dobutamine stress echocardiography is a time-consuming test, often requiring atropine at the end of the protocol to achieve target heart rate (HR). We examined whether earlier administration of atropine in appropriate patients would shorten test time and increase the likelihood of achieving peak HR. METHODS: Two hundred and seventy consecutive patients were randomized prospectively to conventional or early atropine protocols. Of these, 120 patients with an inadequate HR response [mid-30 microg/kg/min HR<100 (age <50) or <90 (age >50); or mid-40 microg/kg/min stage HR<120 (age <50) or <110 (age >50)] were included in the analysis. The remaining patients were used in a model to define which patients are likely to require atropine. RESULTS: The 61 patients receiving early-atropine had decreased test-time relative to the 59 not receiving early atropine (17:05 vs. 18:24 min:sec, P=0.014) accompanied by a 10% reduction in total dobutamine dose (P=0.008). Their HR at end of 40 microg/kg/min was 123+/-18 vs. 105+/-17 respectively, P<0.0001. Only 7% of the early-atropine group failed to reach target HR vs. 15% not receiving early-atropine. By multivariate analysis, age (P<0.0001), HR at end of 30 microg/kg/min stage (P<0.0001), beta blocker use (P=0.009) and baseline HR (P=0.04) were predictors of need for atropine. CONCLUSION: Giving atropine early in appropriate patients can reduce test times without an increase in side effects. Our model enables accurate prediction of these patients. PMID- 11916604 TI - Left ventricular volume analysis for the detection of coronary artery disease during dobutamine stress echocardiography in patients undergoing vascular surgery. AB - AIMS: The purpose of the study was to prospectively evaluate the effectiveness of left ventricular volume changes analysis as compared to wall motion assessment for detecting coronary artery disease during dobutamine stress echocardiography in patients undergoing elective vascular surgery. METHODS AND RESULTS: Left ventricular volumes, measured by using the ellipsoid biplane method combining the apical four- and two-chamber echocardiographic views, and classical wall motion score were determined at rest and peak stress (dobutamine infusion 5-40 microg/kg/min+/- atropine 0.25-1mg) in 68 consecutive patients. A positive test was defined as a decrease of less than 15% in left ventricular end-diastolic or end-systolic volume at peak stress for volume analysis and as an increase in score between rest and peak stress in one or more segments for wall motion assessment. Stress test was not analysable in five patients. Coronary angiography revealed significant coronary artery disease (coronary stenosis >or=70%) in 28/63 (44%) patients: one-vessel in 15, two- or three-vessel disease in 13. Overall sensitivity and specificity for coronary artery disease detection were 56% and 97% with left ventricular volume analysis, as compared to 64% and 89% with wall motion assessment. For patients with two- or three-vessel disease, sensitivity and specificity of volume analysis reached 92%. CONCLUSION: The present data suggest that left ventricular volume change analysis during dobutamine stress echocardiography could be a reliable method for the detection of extensive coronary artery disease for patients undergoing vascular surgery. PMID- 11916605 TI - Sensitivity of the Doppler rate of pressure rise to changes in the inotropic state: an experimental comparison with invasively obtained dP/dt. AB - AIMS: To investigate the sensitivity of the rate of pressure rise obtained by Doppler to changes in the inotropic state by comparing it to simultaneous invasive measurements of dP/dt under different conditions of contractility. METHODS AND RESULTS: Mitral regurgitation was provoked in five pigs, and simultaneous measurements of dP/dt and the Doppler-estimated rate of pressure rise were made with a micro-manometer and with continuous-wave Doppler. Changes in the inotropic state were induced by drug infusion and by ischaemia. One hundred and twenty-seven simultaneous measurements were made with a correlation coefficient between the Doppler-estimated rate of pressure rise and dP/dt of 0.85 (P<0.001). Sensitivity to inotropic changes was estimated as the percentage change of each parameter in each condition of contractility, and showed that the Doppler-estimated rate of pressure rise had better sensitivity than dP/dt. CONCLUSION: The sensitivity of Doppler-estimated rate of pressure rise to changes in the inotropic state is greater than that of dP/dt. The correlation between the rate of pressure rise obtained by Doppler and dP/dt is maintained even in extreme conditions of contractility. Therefore, the rate of pressure rise can be considered a good parameter to assess linear changes of contractility. PMID- 11916606 TI - Percutaneous septal ablation for hypertrophic cardiomyopathy and mid-ventricular obstruction. AB - AIMS: Percutaneous transluminal septal myocardial ablation by alcohol-induced septal branch occlusion is a new treatment option in symptomatic patients with hypertrophic cardiomyopathy and subaortic, SAM-associated obstruction. We report on a patient with mid-ventricular obstruction and echocardiographic-guided reduction of septal hypertrophy. METHODS AND RESULTS: A 52-year-old woman with NYHA class III and recurrent exercise-induced syncope suffered from hypertrophic cardiomyopathy with mid-ventricular obstruction. She had a systolic gradient of 71 mmHg at rest and 153 mmHg post-extrasystole, and diastolic inflow gradient of 20 mmHg. Echo-guided percutaneous transluminal septal myocardial ablation with occlusion of the fourth septal branch resulted in acute reduction and final elimination of systolic, as well as diastolic resting and provocable gradients. Complications were not seen. At 3 months' follow-up the patient was asymptomatic and without further syncopes. CONCLUSIONS: Echocardiographic-guided percutaneous transluminal septal myocardial ablation is able to reduce gradients in hypertrophic cardiomyopathy and mid-ventricular obstruction with consecutive improvement of symptoms. PMID- 11916608 TI - A closed interatrial septum aneurysm, filled with blood, mimicking a tumour in the right atrium. AB - We report the case of a 70-year-old woman with rheumatic mitral stenosis and a transient ischaemic attack. Transoesophageal echocardiography revealed a cystic mass in the right atrium, hanging to the interatrial septum by a pedicle, not circulating. The mass was heterogeneous and suggested a tumour (myxoma) or a thrombus. Surgical resection showed it was an interatrial septal aneurysm, closed on itself, filled with blood. The usual causes of cardiac tumours and pathogeny of large interatrial aneurysms are discussed. PMID- 11916607 TI - Involvement of right ventricle in left ventricular hypertrophic cardiomyopathy: analysis by pulsed Doppler tissue imaging. AB - AIMS: This study uses pulsed Doppler tissue imaging to analyse right ventricular myocardial function and its interaction with left ventricle in hypertrophic cardiomyopathy involving ventricular septum. METHODS AND RESULTS: Thirty-four patients with septal hypertrophic cardiomyopathy and 30 normal subjects, comparable for sex, age, body mass index and heart rate, underwent complete standard Doppler echocardiography and pulsed Doppler tissue imaging of both posterior septum and right ventricular free wall, calculating myocardial velocities and both systolic and diastolic time intervals. Except for peak velocity A, the other Doppler tricuspid inflow measurements were significantly impaired in hypertrophic cardiomyopathy, without changes of tricuspid annular systolic excursion. Right ventricular Doppler tissue imaging showed longer right ventricular myocardial relaxation time in hypertrophic cardiomyopathy than in controls (P<0.00001), without a significant difference from other myocardial diastolic and systolic measurements. In the overall population, Doppler measurements of right and left ventricular inflow were not significantly associated, while (with the exception of myocardial deceleration time) all the other myocardial systolic and diastolic measurements derived by tissue imaging were directly related to the homologous septal myocardial indexes. In addition, a significant inverse relation was found between septal wall thickness and myocardial relaxation index (right-left myocardial relaxation time/right ventricular relaxation time x 100). CONCLUSIONS: This study shows the usefulness of pulsed Doppler tissue imaging to detect impairment of right ventricular myocardial function and to provide evidence about ventricular interaction in forms of hypertrophic cardiomyopathy which involve interventricular septum. PMID- 11916609 TI - Non-bacterial thrombotic endocarditis. AB - A 63-year-old man presented with cachexia and confusion. He was found to have culture-negative endocarditis affecting his aortic valve. Despite treatment with broad-spectrum antibiotics and extensive investigation for an underlying cause, he suffered a large cerebral infarct and died. At post-mortem he was found to have non-bacterial thrombotic endocarditis and a metastatic signet-ring carcinoma. PMID- 11916611 TI - Graves' ophthalmopathy: a preventable disease? AB - Most patients with Graves' disease have some degree of ocular involvement, but only 3-5% of them develop severe ophthalmopathy (1). The reasons why only such a minority of patients with Graves' disease have severe expression of the ophthalmopathy remain to be elucidated. One possible explanation is that non severe ophthalmopathy and severe ophthalmopathy are two different disorders with different genetic backgrounds; alternatively, they might be part of a spectrum of different conditions ranging from absent ocular involvement to most severe ophthalmopathy. In this case, external variables (i.e. environmental factors) must contribute to the nature of the expression of the disease. How important are they? How far can our intervention on environmental factors go towards reducing the risk of progression of the ophthalmopathy? In other words, to which extent, if any, is Graves' ophthalmopathy preventable? The aim of this mini-review is to address the above issues. PMID- 11916612 TI - T-box and isolated ACTH deficiency. PMID- 11916613 TI - The role of IGF-I in the development of cardiovascular disease in type 2 diabetes mellitus: is prevention possible? AB - The incidence of peripheral, cerebro- and cardiovascular disease (CVD) in patients with type 2 diabetes mellitus is approximately twice as high as in the non-diabetic population. Conventional cardiovascular risk factors such as plasma lipids, lipoproteins and hypertension only partially explain this excessive risk of developing atherosclerosis and CVD. Meta-analysis of studies performed in non diabetic populations indicates that the risk of CVD increases continuously with glucose levels above 4.2 mmol/l. The glucose hypothesis suggests that treatment which normalizes glucose levels prevents or delays the long-term complications of diabetes mellitus. However, the outcome of the UK Prospective Diabetes Study demonstrates that glucose control does not completely prevent CVD.In healthy subjects, serum IGF-I levels peak in early adulthood, after which they gradually decrease with increasing age. Several observations suggest that there is a premature and progressive age-related decline in serum IGF-I bioactivity in type 2 diabetics, which eventually results in a (relative) IGF-I deficiency. In type 2 diabetics, close relationships have been demonstrated between glycaemic control and serum IGF-I levels, with worse control being associated with lower IGF-I levels. Several studies (in non-diabetics) suggest that lowered circulating IGF-I levels account for a poor outcome of CVD. We previously observed in a population based study that a genetically determined lowered IGF-I expression increases the risk of myocardial infarction with type 2 diabetes. This genetic approach overcomes the problem that cross-sectional studies cannot distinguish whether changes in IGF-I levels are a cause or a consequence of a disease. IGF-I is an important metabolic regulatory hormone. In addition, IGF-I suppresses myocardial apoptosis and improves myocardial function in various models of experimental cardiomyopathy. Compared with other growth factors, the 'survival' effect of IGF I on myocardium seems rather unique.Therefore, we hypothesize that the premature and progressive decline in serum IGF-I bioactivity in ageing patients with type 2 diabetics is an important pathophysiological abnormality. It contributes not only to elevated glucose and lipid levels, but also to the progression and the poor outcome of CVD. If this hypothesis is proven to be right, treatment with IGF-I as an adjunct to insulin offers great potential and might not only improve metabolic control but also reduce the incidence and prevalence of CVD in type 2 diabetes patients. However, there is as yet no experimental evidence that long-term (replacement) treatment with IGF-I prevents, delays or reduces CVD in type 2 diabetes patients. Clinical trials are necessary to prove that long-term IGF-I treatment, preferably in the form of a better-tolerated IGF-I/IGF-binding protein 3 complex, improves the overall cardiovascular risk in type 2 diabetes. PMID- 11916614 TI - Markers of potential coeliac disease in patients with Hashimoto's thyroiditis. AB - OBJECTIVE: Coeliac disease (CD) is associated with autoimmune thyroid disease. Gluten sensitivity represents a spectrum, with at one end cases with severe gluten-dependent enteropathy, and at the other subjects with minor signs of deranged mucosal immune response. The aim of this paper was to look for signs of minor small bowel injury and immunohistochemical markers of gluten sensitivity in a group of patients with Hashimoto's disease. SUBJECTS AND METHODS: Fourteen patients with Hashimoto's thyroiditis without serological evidence of CD underwent immunohistochemical analysis of jejunal biopsies. RESULTS: In 6/14 cases (43%) an increased density of gammadelta T cell receptor bearing intra epithelial lymphocytes was found. In 6/14 (43%) signs of mucosal T cell activation (presence of interleukin 2 (IL2) receptor (CD25) on lamina propria T cells and/or expression of human lymphocyte antigen (HLA)-DR molecules on crypt epithelial cells) were noted. In 4 out of 6 such cases, HLA haplotypes were described in association with CD. CONCLUSION: A significant proportion of patients with Hashimoto's thyroiditis present signs of 'potential' CD and of activated mucosal T cell immunity. The gluten dependence of such findings remains to be ascertained. PMID- 11916615 TI - Two novel variants in the thyroxine-binding globulin (TBG) gene behind the diagnosis of TBG deficiency. AB - OBJECTIVE: Search for germline mutations in the thyroxine-binding globulin (TBG) gene of two unrelated Portuguese females of Caucasian origin in whom the diagnosis of TBG deficiency was suspected because of suppressed TSH despite marginally low total thyroxine and tri-iodothyronine. DESIGN AND METHODS: Screening for germline mutations was conducted by non-radioactive PCR-SSCP analysis. The variants documented by this approach were characterized by sequencing. Moreover, in order to define whether they were mutations or polymorphisms we looked for the same variants analysing 100 alleles at random. To achieve this goal we used, alternatively, restriction analysis and the minisequencing method with an automated capillary electrophoresis system and fluorescent-labelled dideoxynucleotides. RESULTS AND CONCLUSIONS: Two novel variants, one in each patient, were identified. One, involved codon 23 (TCA- >TAA) and the other, codon 223 (CAA-->TAA). Analysis of 50 DNA samples, randomly chosen, revealed that all were homozygous for the wild variant at codon 23. One of them was heterozygous for the variant CAA-->TAA at codon 223. This sample was found to correspond to a Caucasian female in whom serum TBG proved to be not detected. Since both variants identified result in stop codons likely to induce truncated TBG proteins, they are probably responsible for the TBG phenotype observed in the individuals studied. PMID- 11916616 TI - Two novel missense mutations in the thyroid peroxidase gene, R665W and G771R, result in a localization defect and cause congenital hypothyroidism. AB - OBJECTIVE: Thyroid peroxidase (TPO) deficiency is one of the causes of thyroid dyshormonogenesis, because TPO plays a key role in thyroid hormone biosynthesis. To determine the frequency and pattern of TPO abnormalities, we have been screening TPO genes of patients with congenital goitrous hypothyroidism. SUBJECTS AND METHODS: TPO genes of a patient with congenital goitrous hypothyroidism and her parents were directly sequenced, and two novel missense mutations (R665W and G771R) were found. The former was derived from her father and the latter from her mother. R665 and G771 were well conserved in the peroxidase superfamily. When mRNAs containing each of the mutations were transfected into CHO-K1 cells, each cell showed faint TPO enzyme activity. However, immunofluorescence and immunoelectron microscopic analyses revealed that neither of the mutated TPOs reached the plasma membrane. CONCLUSIONS: Two novel missense mutations in the TPO gene were found. TPO proteins encoded by these mutated alleles showed abnormal cellular localization; namely, localization on the plasma membrane was disturbed. The loss of plasma membrane localization in mutated TPOs brought about the iodide organification defect, which was diagnosed as congenital hypothyroidism. PMID- 11916617 TI - Craniofacial and brain abnormalities in Laron syndrome (primary growth hormone insensitivity). AB - OBJECTIVE: To investigate abnormalities in the craniofacial structures and in the brain in patients with Laron syndrome. DESIGN: Eleven patients with classical Laron syndrome, nine untreated adults aged 36-68 years and two children aged 4 and 9 years (the latter treated by IGF-I), were studied. METHODS: Magnetic resonance images of the brain were obtained in all the patients. One patient also underwent computed tomography. The maximal diameter of the maxillary and frontal sinuses was measured and compared with reference values, the size of the sphenoid sinus was evaluated in relation to the sella, and the mastoids were evaluated qualitatively (small or normal). The brain was evaluated for congenital anomalies and parenchymal lesions. RESULTS: In the adult untreated patients, the paranasal sinuses and mastoids were small; in six patients, the bone marrow in the base of the skull was not mature. The diploe of the calvaria was thin. On computed tomography in one adult patient, the sutures were still open. A minimal or mild degree of diffuse brain parenchymal loss was seen in ten patients. One patient demonstrated a lacunar infarct and another periventricular high signals on T2 weighted images. Two patients had cerebellar atrophy. CONCLUSIONS: The present study has demonstrated the important role IGF-I plays in the development of the brain and bony structures of the cranium. PMID- 11916619 TI - A randomised controlled clinical trial of antibiotic impregnation of testosterone pellet implants to reduce extrusion rate. AB - OBJECTIVE: Testosterone pellet implantation is a safe, effective and convenient form of depot androgen replacement, with extrusion of pellets following about 10% of procedures the most frequent adverse effect. This study aimed to determine whether extrusion rate could be reduced by antibiotic impregnation of pellets immediately prior to implantation. DESIGN: Prospective, randomised, parallel group, open-label study design in a single centre. One hundred and eighty-six androgen-deficient men (400 implantation procedures) were randomised into either a group who had their pellets soaked for approximately 2 min in gentamicin solution prior to implantation, or a control group who had the standard implantation procedure. METHODS: Extrusion, infection and/or bruising were evaluated prospectively by self-report from the participants, and retrospectively at subsequent implantation. Other variables (site, shaving, skin preparation, operator, pellet batch, bruising) were collected at implantation time. RESULTS: The extrusion rate was 20% lower (odds ratio=0.80, 95% confidence interval (CI) 0.40-1.62) but not statistically different between the two groups (extrusion rate 23/205 (11.2%) for the control group vs 18/195 (9.2%) for the antibiotic-soak group, P=0.42). One operator experienced more total (P=0.0002) and infection related (P=0.0008) extrusions and marginally more bruising (P=0.06) than other operators. The operator effect did not appear to be explained by differences in experience or implantation style. There was a 4.6-fold excess (95% CI 1.6-18.6) of multiple (19 vs 4 expected) over single (22 vs 10 expected) and no (359 vs 386 expected) extrusions. Extrusion was not related to batch number (P=0.15), location (P=0.15), shaving (P=0.32), old or new site (P=0.59), or the presence of suppuration or not (P=0.42); however, povidone-iodine skin disinfectant had statistically fewer extrusions than mixed alcohol solution. CONCLUSIONS: Antibiotic impregnation prior to implantation does not significantly decrease testosterone pellet extrusion rate. An operator effect, not due to experience or procedural style, an excess of multiple extrusions and disinfectant effects were confirmed. Neither location, nor preparation of the site, nor pellet batch, influences extrusion rate. PMID- 11916618 TI - Testosterone substitution of hypogonadal men prevents the age-dependent increases in body mass index, body fat and leptin seen in healthy ageing men: results of a cross-sectional study. AB - INTRODUCTION: In healthy men, body weight and total fat content increase with advancing age, while serum testosterone levels decrease. In order to elucidate whether a causal relationship between these phenomena exists, we investigated the influence of testosterone or human chorionic gonadotrophin substitution on body mass index (BMI), total fat mass and serum leptin in testosterone-treated and untreated hypogonadal patients in comparison with ageing eugonadal men. METHODS: In a cross-sectional study, the inter-relationships of body weight, total fat mass, serum sex hormones and leptin were analysed in untreated hypogonadal men (n=24; age 19-65 years), treated hypogonadal men (n=61; age 20-67 years) and healthy eugonadal men (n=60; age 24-78 years). Total fat mass was assessed by bioimpedance measurement. Univariate and multiple linear regression analysis was used to detect possible differences. RESULTS: In eugonadal men, serum testosterone levels decreased with advancing age (correlation coefficients: r= 0.71; P<0.0001), while BMI (r=0.39; P=0.002), total fat content (r=0.51; P<0.0001) and leptin (r=0.48; P<0.0001) increased significantly. In untreated hypogonadal patients, an increase in BMI (r=0.50; P=0.013) and total fat mass (r=0.41; P=0.044) was also observed with advancing age. However, in substituted hypogonadal patients, no age-dependent change in BMI (r=0.067; P=0.606), body fat content (r=-0.083; P=0.522), serum testosterone (r=-0,071; P=0.59) or serum leptin (r=-0.23; P=0.176) was found. CONCLUSION: Since testosterone-substituted older hypogonadal men show BMI and fat mass similar to those of younger eugonadal men and since non-treated hypogonadal men are similar to normal ageing men, testosterone appears to be an important factor contributing to these changes. Thus ageing men should benefit from testosterone substitution as far as body composition is concerned. PMID- 11916620 TI - Mutational analysis of the autoimmune regulator (AIRE) gene in sporadic autoimmune Addison's disease can reveal patients with unidentified autoimmune polyendocrine syndrome type I. AB - OBJECTIVE: To investigate whether patients with Addison's disease and polyendocrine syndromes have undiagnosed autoimmune polyendocrine syndrome type I (APS I). MATERIALS AND METHODS: Forty patients with clinical manifestations resembling APS I and with autoantibodies typical of this condition were screened for Norwegian autoimmune regulator (AIRE) gene mutations. RESULTS: A 30-year old man who had developed Addison' s disease at the age of 12, but had no other components of APS I, was homozygous for the 1094-1106 deletion mutation in exon 8 of the AIRE gene, the most common mutation found in Norway. CONCLUSIONS: APS I patients with milder and atypical phenotypes are difficult to diagnose on clinical grounds. Autoantibody analysis and mutational analysis of AIRE may therefore be helpful modalities for identifying these individuals. PMID- 11916621 TI - Primary aldosteronism in normokalemic patients with adrenal incidentalomas. AB - OBJECTIVE: Since primary aldosteronism has been reported in asymptomatic incidental adrenal masses (adrenal incidentalomas, AI), the aim of our study was to detect primary aldosteronism in normokalemic patients with AI and to verify whether a raised plasma aldosterone (ALD)/plasma renin activity (PRA) ratio may be useful for diagnosis. DESIGN: One-hundred and twenty-five normokalemic patients with solid AI (90 hypertensives and 35 normotensives) and 82 essential hypertensives (EH) were studied. Upright ALD and PRA determination was performed in all cases while patients with abnormal ALD/PRA ratios were submitted to confirmatory tests (saline infusion and captopril tests) for diagnosis of primary aldosteronism. METHODS: ALD and PRA were measured by specific radioimmunoassays. RESULTS: PRA values in AI hypertensives (1.05+/-0.13 ng/ml/h) were lower than in AI normotensives (1.14+/-0.14 ng/ml/h, P<0.05) and in EH (1.68+/-0.15 ng/ml/h, P<0.0001). The ALD/PRA ratio in AI hypertensives (46.4+/-5.1) was higher than in AI normotensives (30.7+/-5.8, P<0.03) and in EH (33.2+/-3.5). Four patients with EH and 2 AI normotensive patients had elevated ALD/PRA ratios but normal responses to the suppressive tests, thus excluding diagnosis of primary aldosteronism. Eight patients with AI and hypertension had a high ALD/PRA ratio, and 7 of these were further studied: in 5 patients diagnosis of primary aldosteronism was well-established by dynamic tests, adrenal vein sampling or by surgery. CONCLUSIONS: Primary aldosteronism in normokalemic patients with incidentally discovered adrenal masses was detected in 4 of all cases and in at least 5.5% of those with hypertension. Consequently, these patients, particularly if hypertensive, need to be routinely studied to exclude this hormonal disease. Evaluation of the ALD/PRA ratio seems to be a simple and reliable test for diagnosis. PMID- 11916622 TI - Bone mineral density and quantitative ultrasound in adults with cystic fibrosis. AB - OBJECTIVE: With increasing life span osteoporosis becomes a more recognized problem in patients with cystic fibrosis (CF). The aim of this cross-sectional study in 75 adult patients with CF (mean age 25.3 years) was to assess the prevalence of low bone mineral density (BMD) by dual-energy x-ray absorptiometry (DEXA) and, for the first time, by quantitative ultrasound (QUS), and to identify predicting factors. DESIGN AND METHODS: Bone status was assessed at the lumbar spine (L2-L4) and the femoral neck by DEXA, and at the calcaneus by QUS (stiffness index). These data were correlated with a variety of clinical and anthropomorphic variables. Biochemical markers of bone turnover such as osteocalcin, bone-specific alkaline phosphatase, crosslinks in urine, 25-hydroxy vitamin D (25-OH vitamin D), parathyroid hormone, calcium and free testosterone were determined by standard assays. RESULTS: The mean BMD T score (+/-s.e.m.) was -1.4+/-0.17 at the lumbar spine, and -0.54+/-0.16 at the femoral neck. The mean T score of the calcaneal stiffness index was -0.83+/-0.19. Based on a lumbar spine T score <-2.5 by DEXA, 27% of the patients had osteoporosis. Multiple regression analysis showed that the forced expiratory volume in one second (FEV1) and the use of oral glucocorticoids were independent predictors of low lumbar spine BMD, whereas body mass index (BMI) and the use of oral glucocorticoids were independent predictors of low femoral neck BMD. The stiffness index correlated moderately with BMD (0.49-0.62, P<0.0001). QUS had a sensitivity and specificity of only 57% and 89% respectively for diagnosing 'osteoporosis' (based on a femoral neck T score <-2.5 by DEXA). Positive and negative predictive values were 36% and 95% respectively. CONCLUSIONS: Low BMD is frequent in adults with CF and is most strongly correlated with disease severity (BMI, FEV1) and the use of glucocorticoids. Calcaneal QUS might help to screen out patients with a normal BMD, but sensitivity and specificity were not sufficiently high to replace DEXA in these patients. PMID- 11916624 TI - Lack of association between peroxisome proliferator-activated receptor-gamma-2 gene variants and the occurrence of coronary heart disease in patients with diabetes mellitus. AB - OBJECTIVE: Recent evidence indicates that peroxisome proliferator-activated receptor-gamma (PPARgamma) is expressed at high levels in foam cells of atherosclerotic lesions, that PPARgamma agonists may directly modulate vessel wall function and that mutations in the PPARgamma-2 gene are associated with a reduced risk of coronary artery disease. METHODS: We investigated whether known variants in the PPARgamma-2 gene are associated with the occurrence of coronary heart disease (CHD) in 365 patients with type 2 diabetes, prospectively characterised for the presence or absence of CHD. The Pro115Gln, Pro12Ala, Pro467Leu, Val290Met mutations and two polymorphisms C478T and C161T of the PPARgamma-2 gene were examined using PCR, denaturing gradient gel electrophoresis and direct sequencing. RESULTS: The distribution of the Pro12Ala, Ala12Ala, C161T and T161T variants was not significantly different between patients with and without CHD, independent of the gender. The Pro12Ala (P=0.011) and the Ala12Ala (P=0.006) variant were associated with a higher body mass index (BMI) compared with the Pro12Pro genotype. A multiple logistic regression analysis introducing the typical risk factors for CHD (age, sex, hypertension, smoking, BMI >26 kg/m2, elevated low density lipoprotein cholesterol and haemoglobin A1c >7%) identified age >60, male gender, hypertension and a higher BMI, but not the PPARgamma-2 variants, as significant risk factors for CHD in our study groups. CONCLUSION: The PPARgamma-2 genotype was not associated with an increased or reduced risk of the occurrence of CHD and can therefore not be regarded as an independent risk factor for CHD in patients with diabetes mellitus. PMID- 11916623 TI - Associations between body mass, leptin, IGF-I and circulating adrenal androgens in children with obesity and premature adrenarche. AB - OBJECTIVE: To explain why adrenal androgens rise with increasing adiposity during childhood, the role of body mass index (BMI), leptin and IGF-I was studied. We also tested whether these parameters contribute to inducing premature adrenarche (PA). DESIGN: In a cross-sectional study, 26 prepubertal obese children were compared with a group of 26 prepubertal children of normal weight, and 30 children under observation for PA were compared with 30 healthy children, matched for gender, bone age and BMI. METHODS: Relative contributions of BMI standard deviation scores (SDS) and height SDS, as well as unbound leptin and IGF-I, to the levels of androgens, dehydroepiandrosterone sulfate (DHEAS) and Delta4 androstenedione (AD) were investigated by means of stepwise regression models. Logarithms of all hormones were standardised for age using residuals of a simple regression analysis, labelled by the suffix '(res)'. RESULTS: In the obese children, height SDS, IGF-I(res,) DHEAS(res) (all P<0.05), leptin(res) (P<0.01), and AD(res) (P=0.07) were higher than in the controls, and covariates were correlated with each other (leptin(res) versus BMI SDS r=0.71, IGF-I(res) versus height SDS r=0.61). In the stepwise regression analysis of control and obese children, BMI SDS explained 26% and leptin(res) explained 12% of the variability of DHEAS(res), but this percentage remained at 26% when both variables were simultaneously introduced into the model. In contrast, IGF-I(res) and BMI SDS alone each accounted for 15% of the variability of AD, and their joint influence accumulated to explain 28% of the variability of AD(res). In PA, neither BMI SDS nor leptin(res) were correlated with the increased androgens. CONCLUSION: Before the onset of gonadal activity in obese and control children, DHEAS levels, to some extent, are explained by BMI and leptin, while IGF-I in addition to BMI in part accounts for AD levels. Enhanced adrenal androgen secretion in children with PA, however, may be explained by parameters other than leptin or BMI. PMID- 11916625 TI - Association studies between the HSD11B2 gene (encoding human 11beta hydroxysteroid dehydrogenase type 2), type 1 diabetes mellitus and diabetic nephropathy. AB - OBJECTIVE: Mutations in the HSD11B2 gene (encoding human 11beta-hydroxysteroid dehydrogenase type 2) explain the syndrome of apparent mineralocorticoid excess where cortisol acts as a mineralocorticoid. A microsatellite marker within the HSD11B2 gene associates with salt sensitivity and hypertension--phenotypes characterising diabetic nephropathy. Here, we evaluate the HSD11B2 gene as a susceptibility locus for diabetic nephropathy. DESIGN: 150 patients with type 1 diabetes and nephropathy (DN), 145 patients with type 1 diabetes with a long duration of non-nephropathy (LDNN) and 151 normal controls were studied. METHODS: We determined allele frequencies for the (CA)n repeat marker within intron I of the HSD11B2 gene. Duration of type 1 diabetes, diabetic status and renal function were recorded. RESULTS: 11 alleles (138-158) for the marker were observed. Allele 152 was significantly increased in controls compared with LDNN (70.5% vs 57.6%, P(c)<0.05 where P(c) is the P value corrected for multiple comparisons) but no difference was observed between DN and LDNN subjects. Allele 154 was significantly increased in the LDNN compared with the DN subjects (15.9% vs 7.0%, P(c)<0.01) but no difference was observed between DN and controls. A greater proportion of subjects carried at least 1 allele <152 in DN compared with control subjects (47.3% vs 28.5%, P(c)<0.01), but no difference was observed in LDNN compared with control and DN subjects. CONCLUSIONS: Weak associations are reported between the HSD11B2 gene, type 1 diabetes mellitus and nephropathy. The increased frequency of HSD11B2 short alleles in the diabetic groups may reflect reduced renal 11beta-hydroxysteroid dehydrogenase type 2 (11beta-HSD2) activity and may, in part, explain the enhanced salt sensitivity observed in patients with type 1 diabetes. PMID- 11916627 TI - Regulation and expression of a renin-angiotensin system in human pancreas and pancreatic endocrine tumours. AB - OBJECTIVE: Evidence exists for the presence of a renin-angiotensin system (RAS) in the pancreas. The aims of this study were to prove the presence of an intrinsic RAS in the human pancreas and to analyse the role of such an RAS in pancreatic endocrine tumours (PETs). METHODS: Gene expression of key RAS components (angiotensinogen and angiotensin II receptors, namely AT1 and AT2) was investigated in human pancreas and in PETs by semi-quantitative RT-PCR and immunohistochemistry. RESULTS: Expression of mRNAs of RAS components was found in human pancreas and in PETs. Data from semi-quantitative RT-PCR analysis demonstrated an increase in the mRNA expression of angiotensinogen and AT2 receptor in PETs when compared with that in normal pancreas. By immunohistochemistry, angiotensinogen protein was predominantly localized in the pancreatic islets while AT1 receptor protein was in the pancreatic ducts. CONCLUSIONS: The data support the notion of the existence of an intrinsic RAS in the human pancreas. It also indicates, for the first time, that such a local pancreatic RAS is subject to regulation by PETs and its significant change may have pathophysiological relevance in patients with PETs. PMID- 11916626 TI - Proinflammatory cytokines inhibit the expression and function of human type I 5' deiodinase in HepG2 hepatocarcinoma cells. AB - OBJECTIVE: The sick euthyroid syndrome in critically ill patients without primary disease of the thyroid gland is characterised by low serum total triiodothyronine (T3), normal to elevated thyroxine (T4), elevated reverse T3 (rT3) and normal TSH levels. The aim of this work was to clarify if impaired T4 and rT3 5' deiodination is an underlying mechanism. DESIGN AND METHODS: We analysed the effect of the human recombinant proinflammatory cytokines interleukin (IL)-6 and IL-1beta, tumour necrosis factor-alpha (TNF-alpha) and interferon-gamma (IFN gamma) on human type I 5'-iodothyronine deiodinase (5'DI) enzyme activity in the human hepatocarcinoma cell line HepG2, i.e. in a homologous human system. Furthermore, we analysed transcriptional effects of the cytokines by transient transfection assays using the luciferase or chloramphenicol acetyltransferase (CAT) reporter genes under the control of 1480 nucleotides of the human 5'DI promoter. RESULTS: IL-6 at 500 pg/ml and TNF-alpha at 25 ng/ml had no significant effect, whereas 100 ng/ml IFN-gamma or 10 ng/ml IL-1beta reduced 5'DI enzyme activity to 77.9 and 59.5% of control values. IFN-gamma did not alter, IL-6 and TNF-alpha moderately decreased (in the case of IL-6 only in the CAT system), and IL-1beta (0.01-10 ng/ml) dose-dependently inhibited 5'DI promoter activity to a minimum of 38.1%. CONCLUSION: IL-1beta inhibited both 5'DI enzyme and promoter activity and, thus, may exert its effect on thyroid hormone metabolism at least partially through direct inhibition of hepatic 5'DI gene transcription. PMID- 11916628 TI - Effect of different growth hormone (GH) mutants on the regulation of GH-receptor gene transcription in a human hepatoma cell line. AB - OBJECTIVE: G to A transition at position 6664 of the growth hormone (GH-1) gene results in the substitution of Arg183 by His (R183H) in the GH protein and causes a new form of autosomal dominant isolated GH deficiency (IGHD type II). The aim of this study was to assess the bioactivity of this R183H mutant GH in comparison with both other GH variants and the 22-kDa GH in terms of GH-receptor gene regulation. DESIGN AND METHODS: The regulation of the GH-receptor gene (GH receptor/GH binding protein, GHR/GHBP) transcription following the addition of variable concentrations (0, 12.5, 25, 50 and 500 ng/ml) of R183H mutant GH was studied in a human hepatoma cell line (HuH7) cultured in a serum-free hormonally defined medium. In addition, identical experiments were performed using either recombinant human GH (22-kDa GH) as a positive control or two GH-receptor antagonists (R77C mutant GH and pegvisomant (B-2036-PEG)) as negative controls. GHR/GHBP mRNA expression was quantitatively assessed by RT-PCR amplification after 0, 1, 3 and 6 h incubation. RESULTS: Following the addition of R183H mutant GH, GHR/GHBP mRNA changed at a similar rate to that seen in experiments where 22 kDa GH was added, indicating equal bioactivity. At all times and concentrations studied, the addition of R77C mutant GH, however, resulted in a significantly lower increase (P<0.001) of GHR/GHBP mRNA concentration compared with that caused by the addition of either 22-kDa GH or R183H mutant GH. Furthermore, in additional experiments, pegvisomant resulted in an absolute block of GHR/GHBP mRNA expression identical to that seen in control experiments where no 22-kDa GH was added at all. CONCLUSIONS: These data indicate that the R183H mutant GH, although causing an autosomal dominant form of IGHD has an identical effect on GHR/GHBP transcription as its wild-type, the 22-kDa GH. This implies that the IGHD caused by the R183H heterozygous mutation of the GH-1 gene is mainly due to a block of its regulated GH secretion. In addition, the R77C-GH variant and pegvisomant have an antagonistic effect at the level of GHR/GHBP transcription. All these data were confirmed by run-on experiments. In addition, these data highlight, as far as the GH variants are concerned, that a mutational alteration within the GH-1 gene might cause short stature also on the basis of an altered secretory pathway. This fact has to be taken into consideration when growth retardation is clinically diagnosed and studied at the molecular level. Secretory pathways and, therefore, cell-biological mechanisms are of importance and have to be considered in future not only at the scientific but also at the clinical level. PMID- 11916629 TI - Aromatase and breast cancer: W39R, an inactive protein. AB - BACKGROUND: Aromatase (CYP19) catalyzes the conversion of androgens into estrogens. It is in particular involved in development, reproduction and breast cancer. One of its polymorphisms, W39R localized in the N-terminal region of CYP19, significantly decreases breast cancer risk among Japanese women and was chosen for this study. In this work, we studied the structure-function relationships between W39R polymorphism and CYP19 enzyme activity. OBJECTIVE: To examine the kinetic properties of the mutant W39R recombinant protein in transfected human cells devoid of steroidogenic activity. METHODS: Expression vectors for the wild-type or the mutated R39 aromatase were transiently transfected into E293 human embryonal kidney cells. The conversions of androstenedione to estrone and of testosterone and nortestosterone to 17beta estradiol were assayed by RIA. Expression of recombinant cDNAs was analyzed by semi-quantitative RT-PCR and immunoblotting. RESULTS: W39R recombinant protein was devoid of aromatase activity whatever the substrate used. This absence of activity was not due to the lack of expression of the recombinant enzyme since the mRNA and protein were detected. CONCLUSION: Our present in vitro study shows that the R39 mutant is unable to synthesize estrogens. This work provides a novel observation, being consistent with the fact that Japanese women with the variant allele (arg) have significantly lower risk of developing a breast tumor. PMID- 11916630 TI - Binding of heparin to human thyroglobulin (Tg) involves multiple binding sites including a region corresponding to a binding site of rat Tg. AB - OBJECTIVE: Binding of thyroglobulin (Tg) to heparin allows efficient Tg interaction with its endocytic receptor, megalin. Rat Tg (rTg) binds to heparin using an exposed carboxyl terminal region (RELPSRRLKRPLPVK, Arg2489-Lys2503) rich in positively charged residues which is, however, not entirely conserved in human Tg (hTg) (Arg2489-Glu2503, REPPARALKRSLWVE). Here, we investigated whether and how this difference affects binding of heparin. DESIGN: To compare binding of heparin to rTg and hTg. To investigate the role of the sequence 2489-2503 using a peptide-based approach. METHODS: Binding of biotin-labeled heparin to rTg, hTg and to Tg peptides was measured in solid phase assays. RESULTS: Heparin bound to rTg with moderately high affinity (K(d): 34.2 nmol/l, K(i): 37.6 nmol/l) and to hTg with lower affinity (K(d): 118 nmol/l, K(i): 480 nmol/l) and to a lower extent. Binding was dose-dependent and saturable, and was reduced by several specific competitors (Tg itself, unlabeled heparin, lactoferrin). Heparin bound to synthetic peptides corresponding to the rat (rTgP) and to the human (hTgP) Tg sequence 2489-2503. Heparin bound to rTgP to a greater extent and with greater affinity than to hTgP. An antibody against hTgP reduced binding of heparin to intact hTg by 30%, suggesting that in hTg this region is, in part, involved in heparin binding, but also that other regions account for most of the binding. Starting from the sequence of rTgP, we designed 6 synthetic 'mutant' peptides by replacing one amino acid residue of rTgP with the corresponding residue of the sequence of hTgP. Heparin bound to 5 of 6 mutant peptides to a lower extent and with lower affinity than to rTgP. CONCLUSIONS: In spite of a reduced binding ability of the sequence 2489-2503, hTg binds to heparin, in part, using alternative, as yet unidentified, binding sites. Substitution of both positive and neutral residues within the sequence 2489-2503 reduced heparin-binding, suggesting that not only charge, but also sequence and/or conformation, may account for the heparin-binding ability of this region of Tg. PMID- 11916631 TI - Can drugs help you lose weight? PMID- 11916632 TI - Hormones and heart disease--down, not out. PMID- 11916633 TI - Is high-normal blood pressure really normal? PMID- 11916635 TI - Heart lines. Glimpsing the future for elderly heart attack survivors. PMID- 11916634 TI - Heart lines. Taking aim at high homocysteine. PMID- 11916636 TI - Ask the Doctor. Someone I work with has lost a lot of weight with a high-protein diet, and he sure seems to enjoy eating all the things my doctor tells me to stay away from! Should I give this diet a try? PMID- 11916638 TI - Probiotics: using bacteria to improve health. PMID- 11916639 TI - Nutrition. Are you getting too much vitamin A? PMID- 11916640 TI - Aging. Getting forgetful: is it dementia? PMID- 11916641 TI - By the way, doctor. As a regular reader of your newsletter, I have a pretty good idea of what a healthy person's cholesterol and triglyceride levels should be. But last year, I had a couple of tests, and the results were completely different. According to the first, my triglycerides were 285, my HDL 31, and my LDL 109. Eight months later, the same measurements came in at 175, 44, and 109. What could cause such a big swing? Is it something I should be worried about? PMID- 11916642 TI - By the way, doctor. I've always used a styptic pencil to stop the bleeding after I cut myself shaving. Recently, I tried it on a small sore on my face that was a little wet and looked infected. It healed up beautifully. Do you think it's safe to use a styptic pencil for that purpose? PMID- 11916643 TI - Depression in children--Part II. PMID- 11916644 TI - Borderline personality: new recommendations. PMID- 11916645 TI - Suicides are not a model. PMID- 11916646 TI - Antidepressants for the heart. PMID- 11916647 TI - Schizophrenia gene discovery. PMID- 11916648 TI - Wine drinking and health. PMID- 11916649 TI - Feeling dizzy. PMID- 11916650 TI - Clinical trials for prostate cancer. PMID- 11916651 TI - Medical memo. Exercise keeps arteries young. PMID- 11916652 TI - On call. My 78-year-old father-in-law may need a bypass to treat his heart problem. My wife read about a new keyhole-type operation. What is it, and would it be better for Dad? PMID- 11916653 TI - Does excess vitamin A cause hip fracture? PMID- 11916655 TI - When anxiety is overwhelming. PMID- 11916654 TI - HRT and Dry Eyes. PMID- 11916656 TI - The genetics of lactose intolerance. PMID- 11916657 TI - Massage is more than an indulgence. PMID- 11916658 TI - By the way, doctor. A few months ago, I was surprised to read that a new study has raised a question about the value of screening mammograms. Is that true? I thought the importance of having such mammograms was well-established. I'm 52 and have been getting annual mammograms for 10 years. Should I stop? PMID- 11916659 TI - The family-3 glycoside hydrolases: from housekeeping functions to host-microbe interactions. PMID- 11916660 TI - Salt stress proteins induced in Listeria monocytogenes. AB - The ability of Listeria monocytogenes to tolerate salt stress is of particular importance, as this pathogen is often exposed to such environments during both food processing and food preservation. In order to understand the survival mechanisms of L. monocytogenes, an initial approach using two-dimensional polyacrylamide gel electrophoresis was performed to analyze the pattern of protein synthesis in response to salt stress. Of 400 to 500 visible proteins, the synthesis of 40 proteins (P < 0.05) was repressed or induced at a higher rate during salt stress. Some of the proteins were identified on the basis of mass spectrometry or N-terminal sequence analysis and database searching. Twelve proteins showing high induction after salt stress were similar to general stress proteins (Ctc and DnaK), transporters (GbuA and mannose-specific phosphotransferase system enzyme IIAB), and general metabolism proteins (alanine dehydrogenase, CcpA, CysK, EF-Tu, Gap, GuaB, PdhA, and PdhD). PMID- 11916661 TI - Application of amplified fragment length polymorphism fingerprinting for taxonomy and identification of the soft rot bacteria Erwinia carotovora and Erwinia chrysanthemi. AB - The soft rot bacteria Erwinia carotovora and Erwinia chrysanthemi are important pathogens of potato and other crops. However, the taxonomy of these pathogens, particularly at subspecies level, is unclear. An investigation using amplified fragment length polymorphism (AFLP) fingerprinting was undertaken to determine the taxonomic relationships within this group based on their genetic relatedness. Following cluster analysis on the similarity matrices derived from the AFLP gels, four clusters (clusters 1 to 4) resulted. Cluster 1 contained Erwinia carotovora subsp. carotovora (subclusters 1a and 1b) and Erwinia carotovora subsp. odorifera (subcluster 1c) strains, while cluster 2 contained Erwinia carotovora subsp. atroseptica (subcluster 2a) and Erwinia carotovora subsp. betavasculorum (subcluster 2b) strains. Clusters 3 and 4 contained Erwinia carotovora subsp. wasabiae and E. chrysanthemi strains, respectively. While E. carotovora subsp. carotovora and E. chrysanthemi showed a high level of molecular diversity (23 to 38% mean similarity), E. carotovora subsp. odorifera, E. carotovora subsp. betavasculorum, E. carotovora subsp. atroseptica, and E. carotovora subsp. wasabiae showed considerably less (56 to 76% mean similarity), which may reflect their limited geographical distributions and/or host ranges. The species- and subspecies-specific banding profiles generated from the AFLPs allowed rapid identification of unknown isolates and the potential for future development of diagnostics. AFLP fingerprinting was also found to be more differentiating than other techniques for typing the soft rot erwinias and was applicable to all strain types, including different serogroups. PMID- 11916662 TI - Role of Bacillus thuringiensis Cry1 delta endotoxin binding in determining potency during lepidopteran larval development. AB - Five economically important crop pests, Manduca sexta, Pieris brassicae, Mamestra brassicae, Spodoptera exigua, and Agrotis ipsilon, were tested at two stages of larval development for susceptibility to Bacillus thuringiensis toxins Cry1Ac, Cry1Ca, Cry1J, and Cry1Ba. Bioassay results for M. sexta showed that resistance to all four Cry toxins increased from the neonate stage to the third-instar stage; the increase in resistance was most dramatic for Cry1Ac, the potency of which decreased 37-fold. More subtle increases in resistance during larval development were seen in M. brassicae for Cry1Ca and in P. brassicae for Cry1Ac and Cry1J. By contrast, the sensitivity of S. exigua did not change during development. At both larval stages, A. ipsilon was resistant to all four toxins. Because aminopeptidase N (APN) is a putative Cry1 toxin binding protein, APN activity was measured in neonate and third-instar brush border membrane vesicles (BBMV). With the exception of S. exigua, APN activity was found to be significantly lower in neonates than in third-instar larvae and thus inversely correlated with increased resistance during larval development. The binding characteristics of iodinated Cry1 toxins were determined for neonate and third instar BBMV. In M. sexta, the increased resistance to Cry1Ac and Cry1Ba during larval development was positively correlated with fewer binding sites in third instar BBMV than in neonate BBMV. The other species-instar-toxin combinations did not reveal positive correlations between potency and binding characteristics. The correlation between binding and potency was inconsistent for the species-instar toxin combinations used in this study, reaffirming the complex mode of action of Cry1 toxins. PMID- 11916663 TI - Activity and diversity of sulfate-reducing bacteria in a petroleum hydrocarbon contaminated aquifer. AB - Microbial sulfate reduction is an important metabolic activity in petroleum hydrocarbon (PHC)-contaminated aquifers. We quantified carbon source-enhanced microbial SO(4)(2-) reduction in a PHC-contaminated aquifer by using single-well push-pull tests and related the consumption of sulfate and added carbon sources to the presence of certain genera of sulfate-reducing bacteria (SRB). We also used molecular methods to assess suspended SRB diversity. In four consecutive tests, we injected anoxic test solutions (1,000 liters) containing bromide as a conservative tracer, sulfate, and either propionate, butyrate, lactate, or acetate as reactants into an existing monitoring well. After an initial incubation period, 1,000 liters of test solution-groundwater mixture was extracted from the same well. Average total test duration was 71 h. We measured concentrations of bromide, sulfate, and carbon sources in native groundwater as well as in injection and extraction phase samples and characterized the SRB population by using fluorescence in situ hybridization (FISH) and denaturing gradient gel electrophoresis (DGGE). Enhanced sulfate reduction concomitant with carbon source degradation was observed in all tests. Computed first-order rate coefficients ranged from 0.19 to 0.32 day(-1) for sulfate reduction and from 0.13 to 0.60 day(-1) for carbon source degradation. Sulfur isotope fractionation in unconsumed sulfate indicated that sulfate reduction was microbially mediated. Enhancement of sulfate reduction due to carbon source additions in all tests and variability of rate coefficients suggested the presence of specific SRB genera and a high diversity of SRB. We confirmed this by using FISH and DGGE. A large fraction of suspended bacteria hybridized with SRB-targeting probes SRB385 plus SRB385-Db (11 to 24% of total cells). FISH results showed that the activity of these bacteria was enhanced by addition of sulfate and carbon sources during push pull tests. However, DGGE profiles indicated that the bacterial community structure of the dominant species did not change during the tests. Thus, the combination of push-pull tests with molecular methods provided valuable insights into microbial processes, activities, and diversity in the sulfate-reducing zone of a PHC-contaminated aquifer. PMID- 11916664 TI - Uric acid is a genuine metabolite of Penicillium cyclopium and stimulates the expression of alkaloid biosynthesis in this fungus. AB - On searching for endogenous, low-molecular-weight effectors of benzodiazepine alkaloid biosynthesis in Penicillium cyclopium uric acid was isolated from ethanolic or autoclaved mycelial extracts of this fungus. The isolation was based on a three-step high-pressure liquid chromatography procedure guided by a microplate bioassay, and uric acid was identified by mass spectrometry and the uricase reaction. Conidiospore suspensions that were treated with this compound during the early phase of outgrowth developed emerged cultures with an enhanced rate of alkaloid production. Uric acid treatment did not increase the in vitro measurable activity of the rate-limiting biosynthetic enzyme, cyclopeptine synthetase. However, these cultures displayed a reduced rate of uptake of the alkaloid precursor L-phenylalanine into the vacuoles of the hyphal cells as assayed in situ. It is suggested that the depressed capacity of vacuolar uptake caused by the contact of outgrowing spores with uric acid liberated from hyphal cells results in an enhanced availability of the precursor L-phenylalanine in the cytoplasm and thus accounts at least in part for the increase in alkaloid production. PMID- 11916665 TI - Induction, isolation, and characterization of two laccases from the white rot basidiomycete Coriolopsis rigida. AB - Previous work has shown that the white rot fungus Coriolopsis rigida degraded wheat straw lignin and both the aliphatic and aromatic fractions of crude oil from contaminated soils. To better understand these processes, we studied the enzymatic composition of the ligninolytic system of this fungus. Since laccase was the sole ligninolytic enzyme found, we paid attention to the oxidative capabilities of this enzyme that would allow its participation in the mentioned degradative processes. We purified two laccase isoenzymes to electrophoretic homogeneity from copper-induced cultures. Both enzymes are monomeric proteins, with the same molecular mass (66 kDa), isoelectric point (3.9), N-linked carbohydrate content (9%), pH optima of 3.0 on 2,6-dimethoxyphenol (DMP) and 2.5 on 2,2'-azinobis(3-ethylbenzothiazoline-6-sulfonic acid) (ABTS), absorption spectrum, and N-terminal amino acid sequence. They oxidized 4-anisidine and numerous phenolic compounds, including methoxyphenols, hydroquinones, and lignin derived aldehydes and acids. Phenol red, an unusual substrate of laccase due to its high redox potential, was also oxidized. The highest enzyme affinity and efficiency were obtained with ABTS and, among phenolic compounds, with 2,6 dimethoxyhydroquinone (DBQH(2)). The presence of ABTS in the laccase reaction expanded the substrate range of C. rigida laccases to nonphenolic compounds and that of MBQH(2) extended the reactions catalyzed by these enzymes to the production of H(2)O(2), the oxidation of Mn(2+), the reduction of Fe(3+), and the generation of hydroxyl radicals. These results confirm the participation of laccase in the production of oxygen free radicals, suggesting novel uses of this enzyme in degradative processes. PMID- 11916666 TI - Cloning of rel from Listeria monocytogenes as an osmotolerance involvement gene. AB - Transposon insertional mutants of Listeria monocytogenes were constructed to identify genes involved in osmotolerance, and one mutant that showed reduced growth under high osmotic pressure was obtained. The cloned gene from the transposon insertion site of the mutant, named rel, was 2,214 bp in length and had very high homology to relA of Bacillus subtilis, which encodes guanosine tetraphosphate (ppGpp) and guanosine pentaphosphate (pppGpp) [collectively designated (p)ppGpp] synthetase during stringent response. The mutant showed a deficiency in (p)ppGpp accumulation. In the parental strain, the amount of intracellular (p)ppGpp was not increased after an osmotic upshift but was slightly decreased compared with the level before the upward shift. The reduced osmotolerance of the mutant was restored to a level almost equal to that of the parent strain when the chromosomal region that included rel of L. monocytogenes was introduced into the mutant. After exposure to methyl glucoside, the rel mutant accumulated (p)ppGpp at a higher level than the basal level and partially restored the ability to grow in NaCl-supplemented brain heart infusion broth. The mutant was found to grow in chemically defined minimal medium supplemented with glycine betaine or carnitine, so-called compatible solutes, and 4% NaCl. Our results suggest that the appropriate intracellular concentration of (p)ppGpp is essential for full osmotolerance in L. monocytogenes and that its mechanism is different from that for the accumulation of compatible solutes. PMID- 11916667 TI - Analysis of bacteria contaminating ultrapure water in industrial systems. AB - Bacterial populations inhabiting ultrapure water (UPW) systems were investigated. The analyzed UPW systems included pilot scale, bench scale, and full size UPW plants employed in the semiconductor and other industries. Bacteria present in the polishing loop of the UPW systems were enumerated by both plate counts and epifluorescence microscopy. Assessment of bacterial presence in UPW by epifluorescence microscopy (cyanotolyl tetrazolium chloride [CTC] and DAPI [4',6' diamidino-2-phenylindole] staining) showed significantly higher numbers (10 to 100 times more bacterial cells were detected) than that determined by plate counts. A considerable proportion of the bacteria present in UPW (50 to 90%) were cells that did not give a positive signal with CTC stain. Bacteria isolated from the UPW systems were mostly gram negative, and several groups seem to be indigenous for all of the UPW production systems studied. These included Ralstonia pickettii, Bradyrhizobium sp., Pseudomonas saccharophilia, and Stenotrophomonas strains. These bacteria constituted a significant part of the total number of isolated strains (>or=20%). Two sets of primers specific to R. pickettii and Bradyrhizobium sp. were designed and successfully used for the detection of the corresponding bacteria in the concentrated UPW samples. Unexpectedly, nifH gene sequences were found in Bradyrhizobium sp. and some P. saccharophilia strains isolated from UPW. The widespread use of nitrogen gas in UPW plants may be associated with the presence of nitrogen-fixing genes in these bacteria. PMID- 11916668 TI - EglC, a new endoglucanase from Aspergillus niger with major activity towards xyloglucan. AB - A novel gene, eglC, encoding an endoglucanase, was cloned from Aspergillus niger. Transcription of eglC is regulated by XlnR, a transcriptional activator that controls the degradation of polysaccharides in plant cell walls. EglC is an 858 amino-acid protein and contains a conserved C-terminal cellulose-binding domain. EglC can be classified in glycoside hydrolase family 74. No homology to any of the endoglucanases from Trichoderma reesei was found. In the plant cell wall xyloglucan is closely linked to cellulose fibrils. We hypothesize that the EglC cellulose-binding domain anchors the enzyme to the cellulose chains while it is cleaving the xyloglucan backbone. By this action it may contribute to the degradation of the plant cell wall structure together with other enzymes, including hemicellulases and cellulases. EglC is most active towards xyloglucan and therefore is functionally different from the other two endoglucanases from A. niger, EglA and EglB, which exhibit the greatest activity towards beta-glucan. Although the mode of action of EglC is not known, this enzyme represents a new enzyme function involved in plant cell wall polysaccharide degradation by A. niger. PMID- 11916669 TI - The phenolic hydroxyl group of carvacrol is essential for action against the food borne pathogen Bacillus cereus. AB - The natural antimicrobial compound carvacrol shows a high preference for hydrophobic phases. The partition coefficients of carvacrol in both octanol-water and liposome-buffer phases were determined (3.64 and 3.26, respectively). Addition of carvacrol to a liposomal suspension resulted in an expansion of the liposomal membrane. Maximum expansion was observed after the addition of 0.50 micromol of carvacrol/mg of L-alpha-phosphatidylethanolamine. Cymene, a biological precursor of carvacrol which lacks a hydroxyl group, was found to have a higher preference for liposomal membranes, thereby causing more expansion. The effect of cymene on the membrane potential was less pronounced than the effect of carvacrol. The pH gradient and ATP pools were not affected by cymene. Measurement of the antimicrobial activities of compounds similar to carvacrol (e.g., thymol, cymene, menthol, and carvacrol methyl ester) showed that the hydroxyl group of this compound and the presence of a system of delocalized electrons are important for the antimicrobial activity of carvacrol. Based on this study, we hypothesize that carvacrol destabilizes the cytoplasmic membrane and, in addition, acts as a proton exchanger, thereby reducing the pH gradient across the cytoplasmic membrane. The resulting collapse of the proton motive force and depletion of the ATP pool eventually lead to cell death. PMID- 11916670 TI - Comparison of extracellular enzyme activities and community composition of attached and free-living bacteria in porous medium columns. AB - Free-living and surface-associated microbial communities in sand-packed columns perfused with groundwater were compared by examination of compositional and functional characteristics. The composition of the microbial communities was assessed by bulk DNA extraction, PCR amplification of 16S ribosomal DNA fragments, separation of these fragments by denaturing gradient gel electrophoresis, and sequence analysis. Community function was assessed by measurement of beta-glucosidase and aminopeptidase extracellular enzyme activities. Free-living populations in the aqueous phase exhibited a greater diversity of phylotypes than populations associated with the solid phase. The attached bacterial community displayed significantly greater beta-glucosidase and aminopeptidase enzyme activities per volume of porous medium than those of the free-living community. On a per-cell basis, the attached community had a significantly higher cell-specific aminopeptidase enzyme activity (1.07 x 10(-7) nmol cell(-1) h(-1)) than the free-living community (5.02 x 10(-8) nmol cell(-1) h(-1)). Conversely, the free-living community had a significantly higher cell specific beta-glucosidase activity (1.92 x 10(-6) nmol cell(-1) h(-1)) than the surface-associated community (6.08 x 10(-7) nmol cell(-1) h(-1)). The compositional and functional differences observed between these two communities may reflect different roles for these distinct but interacting communities in the decomposition of natural organic matter or biodegradation of xenobiotics in aquifers. PMID- 11916671 TI - Phylogenetic diversity of marine cyanophage isolates and natural virus communities as revealed by sequences of viral capsid assembly protein gene g20. AB - In order to characterize the genetic diversity and phylogenetic affiliations of marine cyanophage isolates and natural cyanophage assemblages, oligonucleotide primers CPS1 and CPS8 were designed to specifically amplify ca. 592-bp fragments of the gene for viral capsid assembly protein g20. Phylogenetic analysis of isolated cyanophages revealed that the marine cyanophages were highly diverse yet more closely related to each other than to enteric coliphage T4. Genetically related marine cyanophage isolates were widely distributed without significant geographic segregation (i.e., no correlation between genetic variation and geographic distance). Cloning and sequencing analysis of six natural virus concentrates from estuarine and oligotrophic offshore environments revealed nine phylogenetic groups in a total of 114 different g20 homologs, with up to six clusters and 29 genotypes encountered in a single sample. The composition and structure of natural cyanophage communities in the estuary and open-ocean samples were different from each other, with unique phylogenetic clusters found for each environment. Changes in clonal diversity were also observed from the surface waters to the deep chlorophyll maximum layer in the open ocean. Only three clusters contained known cyanophage isolates, while the identities of the other six clusters remain unknown. Whether or not these unidentified groups are composed of bacteriophages that infect different Synechococcus groups or other closely related cyanobacteria remains to be determined. The high genetic diversity of marine cyanophage assemblages revealed by the g20 sequences suggests that marine viruses can potentially play important roles in regulating microbial genetic diversity. PMID- 11916672 TI - Temporal changes in archaeal diversity and chemistry in a mid-ocean ridge subseafloor habitat. AB - The temporal variation in archaeal diversity in vent fluids from a midocean ridge subseafloor habitat was examined using PCR-amplified 16S rRNA gene sequence analysis and most-probable-number (MPN) cultivation techniques targeting hyperthermophiles. To determine how variations in temperature and chemical characteristics of subseafloor fluids affect the microbial communities, we performed molecular phylogenetic and chemical analyses on diffuse-flow vent fluids from one site shortly after a volcanic eruption in 1998 and again in 1999 and 2000. The archaeal population was divided into particle-attached (>3-microm diameter cells) and free-living fractions to test the hypothesis that subseafloor microorganisms associated with active hydrothermal systems are adapted for a lifestyle that involves attachment to solid surfaces and formation of biofilms. To delineate between entrained seawater archaea and the indigenous subseafloor microbial community, a background seawater sample was also examined and found to consist only of Group I Crenarchaeota and Group II Euryarchaeota, both of which were also present in vent fluids. The indigenous subseafloor archaeal community consisted of clones related to both mesophilic and hyperthermophilic Methanococcales, as well as many uncultured Euryarchaeota, some of which have been identified in other vent environments. The particle-attached fraction consistently showed greater diversity than the free-living fraction. The fluid and MPN counts indicate that while culturable hyperthermophiles represent less than 1% of the total microbial community, the subseafloor at new eruption sites does support a hyperthermophilic microbial community. The temperature and chemical indicators of the degree of subseafloor mixing appear to be the most important environmental parameters affecting community diversity, and it is apparent that decreasing fluid temperatures correlated with increased entrainment of seawater, decreased concentrations of hydrothermal chemical species, and increased incidence of seawater archaeal sequences. PMID- 11916674 TI - Reduced oxidative pentose phosphate pathway flux in recombinant xylose-utilizing Saccharomyces cerevisiae strains improves the ethanol yield from xylose. AB - In recombinant, xylose-fermenting Saccharomyces cerevisiae, about 30% of the consumed xylose is converted to xylitol. Xylitol production results from a cofactor imbalance, since xylose reductase uses both NADPH and NADH, while xylitol dehydrogenase uses only NAD(+). In this study we increased the ethanol yield and decreased the xylitol yield by lowering the flux through the NADPH producing pentose phosphate pathway. The pentose phosphate pathway was blocked either by disruption of the GND1 gene, one of the isogenes of 6-phosphogluconate dehydrogenase, or by disruption of the ZWF1 gene, which encodes glucose 6 phosphate dehydrogenase. Decreasing the phosphoglucose isomerase activity by 90% also lowered the pentose phosphate pathway flux. These modifications all resulted in lower xylitol yield and higher ethanol yield than in the control strains. TMB3255, carrying a disruption of ZWF1, gave the highest ethanol yield (0.41 g g( 1)) and the lowest xylitol yield (0.05 g g(-1)) reported for a xylose-fermenting recombinant S. cerevisiae strain, but also an 84% lower xylose consumption rate. The low xylose fermentation rate is probably due to limited NADPH-mediated xylose reduction. Metabolic flux modeling of TMB3255 confirmed that the NADPH-producing pentose phosphate pathway was blocked and that xylose reduction was mediated only by NADH, leading to a lower rate of xylose consumption. These results indicate that xylitol production is strongly connected to the flux through the oxidative part of the pentose phosphate pathway. PMID- 11916673 TI - Molecular method to assess the diversity of Burkholderia species in environmental samples. AB - In spite of the importance of many members of the genus Burkholderia in the soil microbial community, no direct method to assess the diversity of this genus has been developed so far. The aim of this work was the development of soil DNA-based PCR-denaturing gradient gel electrophoresis (DGGE), a powerful tool for studying the diversity of microbial communities, for detection and analysis of the Burkholderia diversity in soil samples. Primers specific for the genus Burkholderia were developed based on the 16S rRNA gene sequence and were evaluated in PCRs performed with genomic DNAs from Burkholderia and non Burkholderia species as the templates. The primer system used exhibited good specificity and sensitivity for the majority of established species of the genus Burkholderia. DGGE analyses of the PCR products obtained showed that there were sufficient differences in migration behavior to distinguish the majority of the 14 Burkholderia species tested. Sequence analysis of amplicons generated with soil DNA exclusively revealed sequences affiliated with sequences of Burkholderia species, demonstrating that the PCR-DGGE method is suitable for studying the diversity of this genus in natural settings. A PCR-DGGE analysis of the Burkholderia communities in two grassland plots revealed differences in diversity mainly between bulk and rhizosphere soil samples; the communities in the latter samples produced more complex patterns. PMID- 11916675 TI - Determination of subunit composition of Clostridium cellulovorans cellulosomes that degrade plant cell walls. AB - Clostridium cellulovorans produces a cellulase enzyme complex (cellulosome). In this study, we isolated two plant cell wall-degrading cellulosomal fractions from culture supernatant of C. cellulovorans and determined their subunit compositions and enzymatic activities. One of the cellulosomal fractions showed fourfold higher plant cell wall-degrading activity than the other. Both cellulosomal fractions contained the same nine subunits (the scaffolding protein CbpA, endoglucanases EngE and EngK, cellobiohydrolase ExgS, xylanase XynA, mannanase ManA, and three unknown proteins), although the relative amounts of the subunits differed. Since only cellobiose was released from plant cell walls by the cellulosomal fractions, cellobiohydrolases were considered to be key enzymes for plant cell wall degradation. PMID- 11916676 TI - Influence of medium buffering capacity on inhibition of Saccharomyces cerevisiae growth by acetic and lactic acids. AB - Acetic acid (167 mM) and lactic acid (548 mM) completely inhibited growth of Saccharomyces cerevisiae both in minimal medium and in media which contained supplements, such as yeast extract, corn steep powder, or a mixture of amino acids. However, the yeast grew when the pH of the medium containing acetic acid or lactic acid was adjusted to 4.5, even though the medium still contained the undissociated form of either acid at a concentration of 102 mM. The results indicated that the buffer pair formed when the pH was adjusted to 4.5 stabilized the pH of the medium by sequestering protons and by lessening the negative impact of the pH drop on yeast growth, and it also decreased the difference between the extracellular and intracellular pH values (Delta(pH)), the driving force for the intracellular accumulation of acid. Increasing the undissociated acetic acid concentration at pH 4.5 to 163 mM by raising the concentration of the total acid to 267 mM did not increase inhibition. It is suggested that this may be the direct result of decreased acidification of the cytosol because of the intracellular buffering by the buffer pair formed from the acid already accumulated. At a concentration of 102 mM undissociated acetic acid, the yeast grew to higher cell density at pH 3.0 than at pH 4.5, suggesting that it is the total concentration of acetic acid (104 mM at pH 3.0 and 167 mM at pH 4.5) that determines the extent of growth inhibition, not the concentration of undissociated acid alone. PMID- 11916678 TI - Comparison of fecal coliform agar and violet red bile lactose agar for fecal coliform enumeration in foods. AB - A 24-h direct plating method for fecal coliform enumeration with a resuscitation step (preincubation for 2 h at 37 +/- 1 degrees C and transfer to 44 +/- 1 degrees C for 22 h) using fecal coliform agar (FCA) was compared with the 24-h standardized violet red bile lactose agar (VRBL) method. FCA and VRBL have equivalent specificities and sensitivities, except for lactose-positive non-fecal coliforms such as Hafnia alvei, which could form typical colonies on FCA and VRBL. Recovery of cold-stressed Escherichia coli in mashed potatoes on FCA was about 1 log unit lower than that with VRBL. When the FCA method was compared with standard VRBL for enumeration of fecal coliforms, based on counting carried out on 170 different food samples, results were not significantly different (P > 0.05). Based on 203 typical identified colonies selected as found on VRBL and FCA, the latter medium appears to allow the enumeration of more true fecal coliforms and has higher performance in certain ways (specificity, sensitivity, and negative and positive predictive values) than VRBL. Most colonies clearly identified on both media were E. coli and H. alvei, a non-fecal coliform. Therefore, the replacement of fecal coliform enumeration by E. coli enumeration to estimate food sanitary quality should be recommended. PMID- 11916677 TI - The ybiT gene of Erwinia chrysanthemi codes for a putative ABC transporter and is involved in competitiveness against endophytic bacteria during infection. AB - We investigated the role in bacterial infection of a putative ABC transporter, designated ybiT, of Erwinia chrysanthemi AC4150. The deduced sequence of this gene showed amino acid sequence similarity with other putative ABC transporters of gram-negative bacteria, such as Escherichia coli and Pseudomonas aeruginosa, as well as structural similarity with proteins of Streptomyces spp. involved in resistance to macrolide antibiotics. The gene contiguous to ybiT, designated as pab (putative antibiotic biosynthesis) showed sequence similarity with Pseudomonas and Streptomyces genes involved in the biosynthesis of antibiotics. A ybiT mutant (BT117) was constructed by marker exchange. It retained full virulence in potato tubers and chicory leaves, but it showed reduced ability to compete in planta against the wild-type strain or against selected saprophytic bacteria. These results indicate that the ybiT gene plays a role in the in planta fitness of the bacteria. PMID- 11916679 TI - Purification and characterization of thermostable endo-1,5-alpha-L-arabinase from a strain of Bacillus thermodenitrificans. AB - A strain of a thermophilic bacterium, tentatively designated Bacillus thermodenitrificans TS-3, with arabinan-degrading activity was isolated. It produced an endo-arabinase (ABN) (EC 3.2.1.99) and two arabinofuranosidases (EC 3.2.1.55) extracellularly when grown at 60 degrees C on a medium containing sugar beet arabinan. The ABN (tentatively called an ABN-TS) was purified 7,417-fold by anion-exchange, hydrophobic, size exclusion, and hydroxyapatite chromatographies. The molecular mass of ABN-TS was 35 kDa as determined by sodium dodecyl sulfate polyacrylamide gel electrophoresis, and the isoelectric point was pH 4.5. The enzyme was observed to be more thermostable than known ABNs; it had a half-life of 4 h at 75 degrees C. The enzyme had optimal activity at 70 degrees C and pH 6.0. The enzyme had apparent K(m) values of 8.5 and 45 mg/ml and apparent V(max) values of 1.6 and 1.1 mmol/min/mg of protein against debranched arabinan (alpha 1,5-arabinan) and arabinan, respectively. The enzyme had no pectin-releasing activity (protopectinase activity) from sugar beet protopectin, differing from an ABN (protopectinase-C) from mesophilic Bacillus subtilis IFO 3134. The pattern of degradation of debranched arabinan by ABN-TS indicated that the enzyme was an endo-acting enzyme and the main end products were arabinobiose and arabinose. The results of preliminary experiments indicated that the culture filtrate of strain TS-3 is suitable for L-arabinose production from sugar beet pulp at high temperature. PMID- 11916680 TI - Proline reverses the abnormal phenotypes of Colletotrichum trifolii associated with expression of endogenous constitutively active Ras. AB - Colletotrichum trifolii is the causative organism of alfalfa anthracnose. We previously cloned and characterized the small prototypical G protein, Ras, of C. trifolii, which is involved in the signaling pathways that mediate interaction between the pathogen and its host. Transformants expressing constitutively active forms of Ras have growth medium-dependent phenotypes. In nutrient-rich media (e.g., yeast extract and peptone), the phenotype of the transformants was indistinguishable from that of the wild type. However, during nutrient starvation, the transformants lose polarity, have distended hyphae, and fail to sporulate and produce appressoria. Since peptone caused the phenotype to revert, amino acids were tested singly and in combination to identify the responsible amino acid(s). We found that 1.6 mM proline in the medium reverses the constitutively active Ras phenotype. PMID- 11916681 TI - Effects of temperature, water activity, and syrup film composition on the growth of Wallemia sebi: development and assessment of a model predicting growth lags in syrup agar and crystalline sugar. AB - We investigated the effects of temperature, water activity (a(w)), and syrup film composition on the CFU growth of Wallemia sebi in crystalline sugar. At a high a(w) (0.82) at both high (20 degrees C) and low (10 degrees C) temperatures, the CFU growth of W. sebi in both white and extrawhite sugar could be described using a modified Gompertz model. At a low a(w) (0.76), however, the modified Gompertz model could not be fitted to the CFU data obtained with the two sugars due to long CFU growth lags and low maximum specific CFU growth rates of W. sebi at 20 degrees C and due to the fact that growth did not occur at 10 degrees C. At an a(w) of 0.82, regardless of the temperature, the carrying capacity (i.e., the cell concentration at t = infinity) of extrawhite sugar was lower than that of white sugar. Together with the fact that the syrup film of extrawhite sugar contained less amino-nitrogen relative to other macronutrients than the syrup film of white sugar, these results suggest that CFU growth of W. sebi in extrawhite sugar may be nitrogen limited. We developed a secondary growth model which is able to predict colony growth lags of W. sebi on syrup agar as a function of temperature and a(w). The ability of this model to predict CFU growth lags of W. sebi in crystalline sugar was assessed. PMID- 11916682 TI - Bacillus stearothermophilus neopullulanase selective hydrolysis of amylose to maltose in the presence of amylopectin. AB - The specificity of Bacillus stearothermophilus TRS40 neopullulanase toward amylose and amylopectin was analyzed. Although this neopullulanase completely hydrolyzed amylose to produce maltose as the main product, it scarcely hydrolyzed amylopectin. The molecular mass of amylopectin was decreased by only one order of magnitude, from approximately 10(8) to 10(7) Da. Furthermore, this neopullulanase selectively hydrolyzed amylose when starch was used as a substrate. This phenomenon, efficient hydrolysis of amylose but not amylopectin, was also observed with cyclomaltodextrinase from alkaliphilic Bacillus sp. strain A2-5a and maltogenic amylase from Bacillus licheniformis ATCC 27811. These three enzymes hydrolyzed cyclomaltodextrins and amylose much faster than pullulan. Other amylolytic enzymes, such as bacterial saccharifying alpha-amylase, bacterial liquefying alpha-amylase, beta-amylase, and neopullulanase from Bacillus megaterium, did not exhibit this distinct substrate specificity at all, i.e., the preference of amylose to amylopectin. PMID- 11916683 TI - Photosynthetic apparatus in Roseateles depolymerans 61A is transcriptionally induced by carbon limitation. AB - Production of a photosynthetic apparatus in Roseateles depolymerans 61A, a recently discovered freshwater beta-Proteobacterium showing characteristics of aerobic phototrophic bacteria, was observed when the cells were subjected to a sudden decrease in carbon sources (e.g., when cells grown with 0.1 to 0.4% Casamino Acids were diluted or transferred into medium containing or=0.2% O(2)), and was reduced in the presence of light. Transcription of the R. depolymerans puf operon is considered to be controlled by changes in carbon nutrients in addition to oxygen tension and light intensity. PMID- 11916684 TI - Microbial diversity of a heavily polluted microbial mat and its community changes following degradation of petroleum compounds. AB - We studied the microbial diversity of benthic cyanobacterial mats inhabiting a heavily polluted site in a coastal stream (Wadi Gaza) and monitored the microbial community response induced by exposure to and degradation of four model petroleum compounds in the laboratory. Phormidium- and Oscillatoria-like cyanobacterial morphotypes were dominant in the field. Bacteria belonging to different groups, mainly the Cytophaga-Flavobacterium-Bacteriodes group, the gamma and beta subclasses of the class Proteobacteria, and the green nonsulfur bacteria, were also detected. In slurry experiments, these communities efficiently degraded phenanthrene and dibenzothiophene completely in 7 days both in the light and in the dark. n-Octadecane and pristane were degraded to 25 and 34% of their original levels, respectively, within 7 days, but there was no further degradation until 40 days. Both cyanobacterial and bacterial communities exhibited noticeable changes concomitant with degradation of the compounds. The populations enriched by exposure to petroleum compounds included a cyanobacterium affiliated phylogenetically with Halomicronema. Bacteria enriched both in the light and in the dark, but not bacteria enriched in any of the controls, belonged to the newly described Holophaga-Geothrix-Acidobacterium phylum. In addition, another bacterial population, found to be a member of green nonsulfur bacteria, was detected only in the bacteria treated in the light. All or some of the populations may play a significant role in metabolizing the petroleum compounds. We concluded that the microbial mats from Wadi Gaza are rich in microorganisms with high biodegradative potential. PMID- 11916685 TI - Specific adhesion to cellulose and hydrolysis of organophosphate nerve agents by a genetically engineered Escherichia coli strain with a surface-expressed cellulose-binding domain and organophosphorus hydrolase. AB - A genetically engineered Escherichia coli cell expressing both organophosphorus hydrolase (OPH) and a cellulose-binding domain (CBD) on the cell surface was constructed, enabling the simultaneous hydrolysis of organophosphate nerve agents and immobilization via specific adsorption to cellulose. OPH was displayed on the cell surface by use of the truncated ice nucleation protein (INPNC) fusion system, while the CBD was surface anchored by the Lpp-OmpA fusion system. Production of both INPNC-OPH and Lpp-OmpA-CBD fusion proteins was verified by immunoblotting, and the surface localization of OPH and the CBD was confirmed by immunofluorescence microscopy. Whole-cell immobilization with the surface anchored CBD was very specific, forming essentially a monolayer of cells on different supports, as shown by electron micrographs. Optimal levels of OPH activity and binding affinity to cellulose supports were achieved by investigating expression under different induction levels. Immobilized cells degraded paraoxon rapidly at an initial rate of 0.65 mM/min/g of cells (dry weight) and retained almost 100% efficiency over a period of 45 days. Owing to its superior degradation capacity and affinity to cellulose, this immobilized cell system should be an attractive alternative for large-scale detoxification of organophosphate nerve agents. PMID- 11916686 TI - NAD(P)H:flavin mononucleotide oxidoreductase inactivation during 2,4,6 trinitrotoluene reduction. AB - Bacteria readily transform 2,4,6-trinitrotoluene (TNT), a contaminant frequently found at military bases and munitions production facilities, by reduction of the nitro group substituents. In this work, the kinetics of nitroreduction were investigated by using a model nitroreductase, NAD(P)H:flavin mononucleotide (FMN) oxidoreductase. Under mediation by NAD(P)H:FMN oxidoreductase, TNT rapidly reacted with NADH to form 2-hydroxylamino-4,6-dinitrotoluene and 4-hydroxylamino 2,6-dinitrotoluene, whereas 2-amino-4,6-dinitrotoluene and 4-amino-2,6 dinitrotoluene were not produced. Progressive loss of activity was observed during TNT reduction, indicating inactivation of the enzyme during transformation. It is likely that a nitrosodinitrotoluene intermediate reacted with the NAD(P)H:FMN oxidoreductase, leading to enzyme inactivation. A half maximum constant with respect to NADH, K(N), of 394 microM was measured, indicating possible NADH limitation under typical cellular conditions. A mathematical model that describes the inactivation process and NADH limitation provided a good fit to TNT reduction profiles. This work represents the first step in developing a comprehensive enzyme level understanding of nitroarene biotransformation. PMID- 11916687 TI - Identification of Listeria monocytogenes genes expressed in response to growth at low temperature. AB - Listeria monocytogenes is a food-borne bacterial pathogen that is able to grow at refrigeration temperatures. To investigate microbial gene expression associated with cold acclimation, we used a differential cDNA cloning procedure known as selective capture of transcribed sequences (SCOTS) to identify bacterial RNAs that were expressed at elevated levels in bacteria grown at 10 degrees C compared to those grown at 37 degrees C. A total of 24 different cDNA clones corresponding to open reading frames in the L. monocytogenes strain EGD-e genome were obtained by SCOTS. These included cDNAs for L. monocytogenes genes involved in previously described cold-adaptive responses (flaA and flp), regulatory adaptive responses (rpoN, lhkA, yycJ, bglG, adaB, and psr), general microbial stress responses (groEL, clpP, clpB, flp, and trxB), amino acid metabolism (hisJ, trpG, cysS, and aroA), cell surface alterations (fbp, psr, and flaA), and degradative metabolism (eutB, celD, and mleA). Four additional cDNAs were obtained corresponding to genes potentially unique to L. monocytogenes and showing no significant similarity to any other previously described genes. Northern blot analyses confirmed increased steady-state levels of RNA for all members of a subset of genes examined during growth at a low temperature. These results indicated that L. monocytogenes acclimation to growth at 10 degrees C likely involves amino acid starvation, oxidative stress, aberrant protein synthesis, cell surface remodeling, alterations in degradative metabolism, and induction of global regulatory responses. PMID- 11916688 TI - Microheterogeneity in 16S ribosomal DNA-defined bacterial populations from a stratified planktonic environment is related to temporal changes and to ecological adaptations. AB - Temporal changes of the bacterioplankton from a meromictic lake (Lake Vilar, Banyoles, Spain) were analyzed with four culture-independent techniques: epifluorescence microscopy, PCR-denaturing gradient gel electrophoresis (DGGE) fingerprinting, fluorescence in situ whole-cell hybridization and flow cytometry sorting. Microscopically, blooms of one cyanobacterium (Synechococcus sp.-like), one green sulfur bacterium (Chlorobium phaeobacteroides-like), and one purple sulfur bacterium (Thiocystis minor-like) were observed at different depths and times. DGGE retrieved these populations and, additionally, populations related to the Cytophaga-Flavobacterium-Bacteroides phylum as predominant community members. The analyses of partial 16S ribosomal DNA sequences from the DGGE fingerprints (550 bp analyzed) revealed higher genetic diversity than expected from microscopic observation for most of these groups. Thus, the sequences of two Synechococcus spp. (both had a similarity of 97% to Synechococcus sp. strain PCC6307 in 16S rRNA), two Thiocystis spp. (similarities to Thiocystis minor of 93 and 94%, respectively), and three Cytophaga spp. (similarities to Cytophaga fermentans of 88 and 89% and to Cytophaga sp. of 93%, respectively) were obtained. The two populations of Synechococcus exhibited different pigment compositions and temporal distributions and their 16S rRNA sequences were 97.3% similar. The two Thiocystis populations differed neither in pigment composition nor in morphology, but their 16S rRNA sequences were only 92.3% similar and they also showed different distributions over time. Finally, two of the Cytophaga spp. showed 96.2% similarity between the 16S rRNA sequences, but one of them was found to be mostly attached to particles and only in winter. Thus, the identity of the main populations changed over time, but the function of the microbial guilds was maintained. Our data showed that temporal shifts in the identity of the predominant population is a new explanation for the environmental 16S rRNA microdiversity retrieved from microbial assemblages and support the hypothesis that clusters of closely related 16S rRNA environmental sequences may actually represent numerous closely related, yet ecologically distinct, populations. PMID- 11916690 TI - Carbon isotope fractionation during aerobic biodegradation of trichloroethene by Burkholderia cepacia G4: a tool to map degradation mechanisms. AB - The strain Burkholderia cepacia G4 aerobically mineralized trichloroethene (TCE) to CO(2) over a time period of approximately 20 h. Three biodegradation experiments were conducted with different bacterial optical densities at 540 nm (OD(540)s) in order to test whether isotope fractionation was consistent. The resulting TCE degradation was 93, 83.8, and 57.2% (i.e., 7.0, 16.2, and 42.8% TCE remaining) at OD(540)s of 2.0, 1.1, and 0.6, respectively. ODs also correlated linearly with zero-order degradation rates (1.99, 1.11, and 0.64 micromol h(-1)). While initial nonequilibrium mass losses of TCE produced only minor carbon isotope shifts (expressed in per mille delta(13)C(VPDB)), they were 57.2, 39.6, and 17.0 per thousand between the initial and final TCE levels for the three experiments, in decreasing order of their OD(540)s. Despite these strong isotope shifts, we found a largely uniform isotope fractionation. The latter is expressed with a Rayleigh enrichment factor, epsilon, and was -18.2 when all experiments were grouped to a common point of 42.8% TCE remaining. Although, decreases of epsilon to -20.7 were observed near complete degradation, our enrichment factors were significantly more negative than those reported for anaerobic dehalogenation of TCE. This indicates typical isotope fractionation for specific enzymatic mechanisms that can help to differentiate between degradation pathways. PMID- 11916689 TI - Effects of growth mode and pyruvate carboxylase on succinic acid production by metabolically engineered strains of Escherichia coli. AB - Escherichia coli NZN111, which lacks activities for pyruvate-formate lyase and lactate dehydrogenase, and AFP111, a derivative which contains an additional mutation in ptsG (a gene encoding an enzyme of the glucose phophotransferase system), accumulate significant levels of succinic acid (succinate) under anaerobic conditions. Plasmid pTrc99A-pyc, which expresses the Rhizobium etli pyruvate carboxylase enzyme, was introduced into both strains. We compared growth, substrate consumption, product formation, and activities of seven key enzymes (acetate kinase, fumarate reductase, glucokinase, isocitrate dehydrogenase, isocitrate lyase, phosphoenolpyruvate carboxylase, and pyruvate carboxylase) from glucose for NZN111, NZN111/pTrc99A-pyc, AFP111, and AFP111/pTrc99A-pyc under both exclusively anaerobic and dual-phase conditions (an aerobic growth phase followed by an anaerobic production phase). The highest succinate mass yield was attained with AFP111/pTrc99A-pyc under dual-phase conditions with low pyruvate carboxylase activity. Dual-phase conditions led to significant isocitrate lyase activity in both NZN111 and AFP111, while under exclusively anaerobic conditions, an absence of isocitrate lyase activity resulted in significant pyruvate accumulation. Enzyme assays indicated that under dual-phase conditions, carbon flows not only through the reductive arm of the tricarboxylic acid cycle for succinate generation but also through the glyoxylate shunt and thus provides the cells with metabolic flexibility in the formation of succinate. Significant glucokinase activity in AFP111 compared to NZN111 similarly permits increased metabolic flexibility of AFP111. The differences between the strains and the benefit of pyruvate carboxylase under both exclusively anaerobic and dual-phase conditions are discussed in light of the cellular constraint for a redox balance. PMID- 11916691 TI - Use of Fe(III) as an electron acceptor to recover previously uncultured hyperthermophiles: isolation and characterization of Geothermobacterium ferrireducens gen. nov., sp. nov. AB - It has recently been recognized that the ability to use Fe(III) as a terminal electron acceptor is a highly conserved characteristic in hyperthermophilic microorganisms. This suggests that it may be possible to recover as-yet uncultured hyperthermophiles in pure culture if Fe(III) is used as an electron acceptor. As part of a study of the microbial diversity of the Obsidian Pool area in Yellowstone National Park, Wyo., hot sediment samples were used as the inoculum for enrichment cultures in media containing hydrogen as the sole electron donor and poorly crystalline Fe(III) oxide as the electron acceptor. A pure culture was recovered on solidified, Fe(III) oxide medium. The isolate, designated FW-1a, is a hyperthermophilic anaerobe that grows exclusively by coupling hydrogen oxidation to the reduction of poorly crystalline Fe(III) oxide. Organic carbon is not required for growth. Magnetite is the end product of Fe(III) oxide reduction under the culture conditions evaluated. The cells are rod shaped, about 0.5 microm by 1.0 to 1.2 microm, and motile and have a single flagellum. Strain FW-1a grows at circumneutral pH, at freshwater salinities, and at temperatures of between 65 and 100 degrees C with an optimum of 85 to 90 degrees C. To our knowledge this is the highest temperature optimum of any organism in the Bacteria. Analysis of the 16S ribosomal DNA (rDNA) sequence of strain FW-1a places it within the Bacteria, most closely related to abundant but uncultured microorganisms whose 16S rDNA sequences have been previously recovered from Obsidian Pool and a terrestrial hot spring in Iceland. While previous studies inferred that the uncultured microorganisms with these 16S rDNA sequences were sulfate-reducing organisms, the physiology of the strain FW-1a, which does not reduce sulfate, indicates that these organisms are just as likely to be Fe(III) reducers. These results further demonstrate that Fe(III) may be helpful for recovering as-yet-uncultured microorganisms from hydrothermal environments and illustrate that caution must be used in inferring the physiological characteristics of at least some thermophilic microorganisms solely from 16S rDNA sequences. Based on both its 16S rDNA sequence and physiological characteristics, strain FW-1a represents a new genus among the Bacteria. The name Geothermobacterium ferrireducens gen. nov., sp. nov., is proposed (ATCC BAA-426). PMID- 11916692 TI - Profiles of airborne fungi in buildings and outdoor environments in the United States. AB - We examined 12,026 fungal air samples (9,619 indoor samples and 2,407 outdoor samples) from 1,717 buildings located across the United States; these samples were collected during indoor air quality investigations performed from 1996 to 1998. For all buildings, both indoor and outdoor air samples were collected with an Andersen N6 sampler. The culturable airborne fungal concentrations in indoor air were lower than those in outdoor air. The fungal levels were highest in the fall and summer and lowest in the winter and spring. Geographically, the highest fungal levels were found in the Southwest, Far West, and Southeast. The most common culturable airborne fungi, both indoors and outdoors and in all seasons and regions, were Cladosporium, Penicillium, nonsporulating fungi, and Aspergillus. Stachybotrys chartarum was identified in the indoor air in 6% of the buildings studied and in the outdoor air of 1% of the buildings studied. This study provides industrial hygienists, allergists, and other public health practitioners with comparative information on common culturable airborne fungi in the United States. This is the largest study of airborne indoor and outdoor fungal species and concentrations conducted with a standardized protocol to date. PMID- 11916693 TI - Identification of quorum-quenching N-acyl homoserine lactonases from Bacillus species. AB - A range of gram-negative bacterial species use N-acyl homoserine lactone (AHL) molecules as quorum-sensing signals to regulate different biological functions, including production of virulence factors. AHL is also known as an autoinducer. An autoinducer inactivation gene, aiiA, coding for an AHL lactonase, was cloned from a bacterial isolate, Bacillus sp. strain 240B1. Here we report identification of more than 20 bacterial isolates capable of enzymatic inactivation of AHLs from different sources. Eight isolates showing strong AHL inactivating enzyme activity were selected for a preliminary taxonomic analysis. Morphological phenotypes and 16S ribosomal DNA sequence analysis indicated that these isolates probably belong to the species Bacillus thuringiensis. Enzymatic analysis with known Bacillus strains confirmed that all of the strains of B. thuringiensis and the closely related species B. cereus and B. mycoides tested produced AHL-inactivating enzymes but B. fusiformis and B. sphaericus strains did not. Nine genes coding for AHL inactivation were cloned either by functional cloning or by a PCR procedure from selected bacterial isolates and strains. Sequence comparison of the gene products and motif analysis showed that the gene products belong to the same family of AHL lactonases. PMID- 11916695 TI - Cyanophycin production in a phycoerythrin-containing marine synechococcus strain of unusual phylogenetic affinity. AB - Thirty-two strains of phycoerythrin-containing marine picocyanobacteria were screened for the capacity to produce cyanophycin, a nitrogen storage compound synthesized by some, but not all, cyanobacteria. We found that one of these strains, Synechococcus sp. strain G2.1 from the Arabian Sea, was able to synthesize cyanophycin. The cyanophycin extracted from the cells was composed of roughly equimolar amounts of arginine and aspartate (29 and 35 mol%, respectively), as well as a small amount of glutamate (15 mol%). Phylogenetic analysis, based on partial 16S ribosomal DNA (rDNA) sequence data, showed that Synechococcus sp. strain G2.1 formed a well-supported clade with several strains of filamentous cyanobacteria. It was not closely related to several other well studied marine picocyanobacteria, including Synechococcus strains PCC7002, WH7805, and WH8018 and Prochlorococcus sp. strain MIT9312. This is the first report of cyanophycin production in a phycoerythrin-containing strain of marine or halotolerant Synechococcus, and its discovery highlights the diversity of this ecologically important functional group. PMID- 11916694 TI - Intracellular carbon fluxes in riboflavin-producing Bacillus subtilis during growth on two-carbon substrate mixtures. AB - Metabolic responses to cofeeding of different carbon substrates in carbon-limited chemostat cultures were investigated with riboflavin-producing Bacillus subtilis. Relative to the carbon content (or energy content) of the substrates, the biomass yield was lower in all cofeeding experiments than with glucose alone. The riboflavin yield, in contrast, was significantly increased in the acetoin- and gluconate-cofed cultures. In these two scenarios, unusually high intracellular ATP-to-ADP ratios correlated with improved riboflavin yields. Nuclear magnetic resonance spectra recorded with amino acids obtained from biosynthetically directed fractional (13)C labeling experiments were used in an isotope isomer balancing framework to estimate intracellular carbon fluxes. The glycolysis-to pentose phosphate (PP) pathway split ratio was almost invariant at about 80% in all experiments, a result that was particularly surprising for the cosubstrate gluconate, which feeds directly into the PP pathway. The in vivo activities of the tricarboxylic acid cycle, in contrast, varied more than twofold. The malic enzyme was active with acetate, gluconate, or acetoin cofeeding but not with citrate cofeeding or with glucose alone. The in vivo activity of the gluconeogenic phosphoenolpyruvate carboxykinase was found to be relatively high in all experiments, with the sole exception of the gluconate-cofed culture. PMID- 11916696 TI - Contribution of Lactococcus lactis cell envelope proteinase specificity to peptide accumulation and bitterness in reduced-fat Cheddar cheese. AB - Bitterness is a flavor defect in Cheddar cheese that limits consumer acceptance, and specificity of the Lactococcus lactis extracellular proteinase (lactocepin) is widely believed to be a key factor in the development of bitter cheese. To better define the contribution of this enzyme to bitterness, we investigated peptide accumulation and bitterness in 50% reduced-fat Cheddar cheese manufactured with single isogenic strains of Lactococcus lactis as the only starter. Four isogens were developed for the study; one was lactocepin negative, and the others produced a lactocepin with group a, e, or h specificity. Analysis of cheese aqueous extracts by reversed-phase high-pressure liquid chromatography confirmed that accumulation of alpha(S1)-casein (f 1-23)-derived peptides f 1-9, f 1-13, f 1-16, and f 1-17 in cheese was directly influenced by lactocepin specificity. Trained sensory panelists demonstrated that Cheddar cheese made with isogenic starters that produced group a, e, or h lactocepin was significantly more bitter than cheese made with a proteinase-negative isogen and that propensity for bitterness was highest in cells that produced group h lactocepin. These results confirm the role of starter proteinase in bitterness and suggest that the propensity of some industrial strains for production of the bitter flavor defect in cheese could be altered by proteinase gene exchange or gene replacement. PMID- 11916697 TI - Development, validation, and application of PCR primers for detection of tetracycline efflux genes of gram-negative bacteria. AB - Phylogenetic analysis of tetracycline resistance genes, which confer resistance due to the efflux of tetracycline from the cell catalyzed by drug:H(+) antiport and share a common structure with 12 transmembrane segments (12-TMS), suggested the monophyletic origin of these genes. With a high degree of confidence, this tet subcluster unifies 11 genes encoding tet efflux pumps and includes tet(A), tet(B), tet(C), tet(D), tet(E), tet(G), tet(H), tet(J), tet(Y), tet(Z), and tet(30). Phylogeny-aided alignments were used to design a set of PCR primers for detection, retrieval, and sequence analysis of the corresponding gene fragments from a variety of bacterial and environmental sources. After rigorous validation with the characterized control tet templates, this primer set was used to determine the genotype of the corresponding tetracycline resistance genes in total DNA of swine feed and feces and in the lagoons and groundwater underlying two large swine production facilities known to be impacted by waste seepage. The compounded tet fingerprint of animal feed was found to be tetCDEHZ, while the corresponding fingerprint of total intestinal microbiota was tetBCGHYZ. Interestingly, the tet fingerprints in geographically distant waste lagoons were identical (tetBCEHYZ) and were similar to the fecal fingerprint at the third location mentioned above. Despite the sporadic detection of chlortetracycline in waste lagoons, no auxiliary diversity of tet genes in comparison with the fecal diversity could be detected, suggesting that the tet pool is generated mainly in the gut of tetracycline-fed animals, with a negligible contribution from selection imposed by tetracycline that is released into the environment. The tet efflux genes were found to be percolating into the underlying groundwater and could be detected as far as 250 m downstream from the lagoons. With yet another family of tet genes, this study confirmed our earlier findings that the antibiotic resistance gene pool generated in animal production systems may be mobile and persistent in the environment with the potential to enter the food chain. PMID- 11916698 TI - Responses of Listeria monocytogenes to acid stress and glucose availability revealed by a novel combination of fluorescence microscopy and microelectrode ion selective techniques. AB - Fluorescence ratio imaging microscopy and microelectrode ion flux estimation techniques were combined to study mechanisms of pH homeostasis in Listeria monocytogenes subjected to acid stress at different levels of glucose availability. This novel combination provided a unique opportunity to measure changes in H(+) at either side of the bacterial membrane in real time and therefore to evaluate the rate of H(+) flux across the bacterial plasma membrane and its contribution to bacterial pH homeostasis. Responses were assessed at external pHs (pH(o)) between 3.0 and 6.0 for three levels of glucose (0, 1, and 10 mM) in the medium. Both the intracellular pH (pH(i)) and net H(+) fluxes were affected by the glucose concentration in the medium, with the highest absolute values corresponding to the highest glucose concentration. In the presence of glucose, the pH(i) remained above 7.0 within a pH(o) range of 4 to 6 and decreased below pH(o) 4. Above pH(o) 4, H(+) extrusion increased correspondingly, with the maximum value at pH(o) 5.5, and below pH(o) 4, a net H(+) influx was observed. Without glucose in the medium, the pH(i) decreased, and a net H(+) influx was observed below pH(o) 5.5. A high correlation (R = 0.75 to 0.92) between the pH(i) and net H(+) flux changes is reported, indicating that the two processes are complementary. The results obtained support other reports indicating that membrane transport processes are the main contributors to the process of pH(i) homeostasis in L. monocytogenes subjected to acid stress. PMID- 11916699 TI - Cloning and characterization of linR, involved in regulation of the downstream pathway for gamma-hexachlorocyclohexane degradation in Sphingomonas paucimobilis UT26. AB - In Sphingomonas paucimobilis UT26, LinD and LinE activities, which are responsible for the degradation of gamma-hexachlorocyclohexane, are inducibly expressed in the presence of their substrates, 2,5-dichlorohydroquinone (2,5 DCHQ) and chlorohydroquinone (CHQ). The nucleotide sequence of the 1-kb upstream region of the linE gene was determined, and an open reading frame (ORF) was found in divergent orientation from linE. Because the putative protein product of the ORF showed similarity to the LysR-type transcriptional regulator (LTTR) family, we named it linR. The fragment containing the putative LTTR recognition sequence (a palindromic TN(11)A sequence), which exists immediately upstream of linE, was ligated with the reporter gene lacZ and was inserted into the plasmid expressing LinR under the control of the lac promoter. When the resultant plasmid was introduced into Escherichia coli, the LacZ activity rose in the presence of 2,5 DCHQ and CHQ. RNA slot blot analysis for the total RNAs of UT26 and UT102, which has an insertional mutation in linR, revealed that the expression of the linD and linE genes was induced in the presence of 2,5-DCHQ, CHQ, and hydroquinone in UT26 but not in UT102. These results indicated that the linR gene is directly involved in the inducible expression of the linD and linE genes. PMID- 11916700 TI - Method for spiking soil samples with organic compounds. AB - We examined the harmful side effects on indigenous soil microorganisms of two organic solvents, acetone and dichloromethane, that are normally used for spiking of soil with polycyclic aromatic hydrocarbons for experimental purposes. The solvents were applied in two contamination protocols to either the whole soil sample or 25% of the soil volume, which was subsequently mixed with 75% untreated soil. For dichloromethane, we included a third protocol, which involved application to 80% of the soil volume with or without phenanthrene and introduction of Pseudomonas fluorescens VKI171 SJ132 genetically tagged with luxAB::Tn5. For both solvents, application to the whole sample resulted in severe side effects on both indigenous protozoa and bacteria. Application of dichloromethane to the whole soil volume immediately reduced the number of protozoa to below the detection limit. In one of the soils, the protozoan population was able to recover to the initial level within 2 weeks, in terms of numbers of protozoa; protozoan diversity, however, remained low. In soil spiked with dichloromethane with or without phenanthrene, the introduced P. fluorescens VKI171 SJ132 was able to grow to a density 1,000-fold higher than in control soil, probably due mainly to release of predation from indigenous protozoa. In order to minimize solvent effects on indigenous soil microorganisms when spiking native soil samples with compounds having a low water solubility, we propose a common protocol in which the contaminant dissolved in acetone is added to 25% of the soil sample, followed by evaporation of the solvent and mixing with the remaining 75% of the soil sample. PMID- 11916701 TI - Genotyping Cryptosporidium parvum with an hsp70 single-nucleotide polymorphism microarray. AB - We investigated the application of an oligonucleotide microarray to (i) specifically detect Cryptosporidium spp., (ii) differentiate between closely related C. parvum isolates and Cryptosporidium species, and (iii) differentiate between principle genotypes known to infect humans. A microarray of 68 capture probes targeting seven single-nucleotide polymorphisms (SNPs) within a 190-bp region of the hsp70 gene of Cryptosporidium parvum was constructed. Labeled hsp70 targets were generated by PCR with biotin- or Cy3-labeled primers. Hybridization conditions were optimized for hybridization time, temperature, and salt concentration. Two genotype I C. parvum isolates (TU502 and UG502), two C. parvum genotype II isolates (Iowa and GCH1), and DNAs from 22 non-Cryptosporidium sp. organisms were used to test method specificity. Only DNAs from C. parvum isolates produced labeled amplicons that could be hybridized to and detected on the array. Hybridization patterns between genotypes were visually distinct, but identification of SNPs required statistical analysis of the signal intensity data. The results indicated that correct mismatch discrimination could be achieved for all seven SNPs for the UG502 isolate, five of seven SNPs for the TU502 isolate, and six of seven SNPs for both the Iowa and GCH1 isolates. Even without perfect mismatch discrimination, the microarray method unambiguously distinguished between genotype I and genotype II isolates and demonstrated the potential to differentiate between other isolates and species on a single microarray. This method may provide a powerful new tool for water utilities and public health officials for assessing point and nonpoint source contamination of water supplies. PMID- 11916702 TI - Metabolic engineering of the morphology of Aspergillus oryzae by altering chitin synthesis. AB - Morphology and alpha-amylase production during submerged cultivation were examined in a wild-type strain (A1560) and in strains of Aspergillus oryzae in which chitin synthase B (chsB) and chitin synthesis myosin A (csmA) have been disrupted (ChsB/G and CM101). In a flowthrough cell, the growth of submerged hyphal elements was studied online, making it possible to examine the growth kinetics of the three strains. The average tip extension rates of the CM101 and ChsB/G strains were 25 and 88% lower, respectively, than that of the wild type. The branching intensity in the CM101 strain was 25% lower than that in the wild type, whereas that in the ChsB/G strain was 188% higher. During batch cultivation, inseparable clumps were formed in the wild-type strain, while no or fewer large inseparable clumps existed in the cultivations of the ChsB/G and CM101 strains. The alpha-amylase productivity was not significantly different in the three strains. A strain in which the transcription of chsB could be controlled by the nitrogen source-regulated promoter niiA (NiiA1) was examined during chemostat cultivation, and it was found that the branching intensity could be regulated by regulating the promoter, signifying an important role for chsB in branching. However, the pattern of branching responded very slowly to the change in transcription, and increased branching did not affect alpha-amylase productivity. alpha-Amylase residing in the cell wall was stained by immunofluorescence, and the relationship between tip number and enzyme secretion is discussed. PMID- 11916703 TI - An exported rhodanese-like protein is induced during growth of Acidithiobacillus ferrooxidans in metal sulfides and different sulfur compounds. AB - By proteomic analysis we found a 21-kDa protein (P21) from Acidithiobacillus ferrooxidans ATCC 19859 whose synthesis was greatly increased by growth of the bacteria in pyrite, thiosulfate, elemental sulfur, CuS, and ZnS and was almost completely repressed by growth in ferrous iron. After we determined the N terminal amino acid sequence of P21, we used the available preliminary genomic sequence of A. ferrooxidans ATCC 23270 to isolate the DNA region containing the p21 gene. The nucleotide sequence of this DNA fragment contained a putative open reading frame (ORF) coding for a 23-kDa protein. This difference in size was due to the presence of a putative signal peptide in the ORF coding for P21. When p21 was cloned and overexpressed in Escherichia coli, the signal peptide was removed, resulting in a mature protein with a molecular mass of 21 kDa and a calculated isoelectric point of 9.18. P21 exhibited 27% identity and 42% similarity to the Deinococcus radiodurans thiosulfate-sulfur transferase (rhodanese; EC 2.8.1.1) and similar values in relation to other rhodaneses, conserving structural domains and an active site with a cysteine, both characteristic of this family of proteins. However, the purified recombinant P21 protein did not show rhodanese activity. Unlike cytoplasmic rhodaneses, P21 was located in the periphery of A. ferrooxidans cells, as determined by immunocytochemical analysis, and was regulated depending on the oxidizable substrate. The genomic context around gene p21 contained other ORFs corresponding to proteins such as thioredoxins and sulfate-thiosulfate binding proteins, clearly suggesting the involvement of P21 in inorganic sulfur metabolism in A. ferrooxidans. PMID- 11916704 TI - Quantification in soil and the rhizosphere of the nematophagous fungus Verticillium chlamydosporium by competitive PCR and comparison with selective plating. AB - A competitive PCR (cPCR) assay was developed to quantify the nematophagous fungus Verticillium chlamydosporium in soil. A gamma-irradiated soil was seeded with different numbers of chlamydospores from V. chlamydosporium isolate 10, and samples were obtained at time intervals of up to 8 weeks. Samples were analyzed by cPCR and by plating onto a semiselective medium. The results suggested that saprophytic V. chlamydosporium growth did occur in soil and that the two methods detected different phases of growth. The first stage of growth, DNA replication, was demonstrated by the rapid increase in cPCR estimates, and the presumed carrying capacity (PCC) of the soil was reached after only 1 week of incubation. The second stage, an increase in fungal propagules presumably due to cell division, sporulation, and hyphal fragmentation, was indicated by a less rapid increase in CFU, and 3 weeks was required to reach the PCC. Experiments with field soil revealed that saprophytic fungal growth was limited, presumably due to competition from the indigenous soil microflora, and that the PCR results were less variable than the equivalent plate count results. In addition, the limit of detection of V. chlamydosporium in field soil was lower than that in gamma irradiated soil, suggesting that there was a background population of the fungus in the field, although the level was below the limit of detection. Tomatoes were infected with the root knot nematode (RKN) or the potato cyst nematode (PCN) along with a PCN-derived isolate of the fungus (V. chlamydosporium isolate Jersey). Increases in fungal growth were observed in the rhizosphere of PCN infested plants but not in the rhizosphere of RKN-infested plants after 14 weeks using cPCR. In this paper we describe for the first time PCR-based quantification of a fungal biological control agent for nematodes in soil and the rhizosphere, and we provide evidence for nematode host specificity that is highly relevant to the biological control efficacy of this fungus. PMID- 11916705 TI - Comparison of soil bacterial communities in rhizospheres of three plant species and the interspaces in an arid grassland. AB - Soil bacteria are important contributors to primary productivity and nutrient cycling in arid land ecosystems, and their populations may be greatly affected by changes in environmental conditions. In parallel studies, the composition of the total bacterial community and of members of the Acidobacterium division were assessed in arid grassland soils using terminal restriction fragment length polymorphism (TRF, also known as T-RFLP) analysis of 16S rRNA genes amplified from soil DNA. Bacterial communities associated with the rhizospheres of the native bunchgrasses Stipa hymenoides and Hilaria jamesii, the invading annual grass Bromus tectorum, and the interspaces colonized by cyanobacterial soil crusts were compared at three depths. When used in a replicated field-scale study, TRF analysis was useful for identifying broad-scale, consistent differences in the bacterial communities in different soil locations, over the natural microscale heterogeneity of the soil. The compositions of the total bacterial community and Acidobacterium division in the soil crust interspaces were significantly different from those of the plant rhizospheres. Major differences were also observed in the rhizospheres of the three plant species and were most apparent with analysis of the Acidobacterium division. The total bacterial community and the Acidobacterium division bacteria were affected by soil depth in both the interspaces and plant rhizospheres. This study provides a baseline for monitoring bacterial community structure and dynamics with changes in plant cover and environmental conditions in the arid grasslands. PMID- 11916706 TI - Staphylococcus aureus growth boundaries: moving towards mechanistic predictive models based on solute-specific effects. AB - The formulation of shelf-stable intermediate-moisture products is a critical food safety issue. Therefore, knowing the precise boundary for the growth-no-growth interface of Staphylococcus aureus is necessary for food safety risk assessment. This study was designed to examine the effects of various humectants and to produce growth boundary models as tools for risk assessment. The molecular mobility and the effects of various physical properties of humectants, such as their glass transition temperatures, their membrane permeability, and their ionic and nonionic properties, on S. aureus growth were investigated. The effects of relative humidity (RH; 84 to 95%, adjusted by sucrose plus fructose, glycerol, or NaCl), initial pH (4.5 to 7.0, adjusted by HCl), and potassium sorbate concentration (0 or 1,000 ppm) on the growth of S. aureus were determined. Growth was monitored by turbidity over a 24-week period. Toxin production was determined by enterotoxin assay. The 1,792 data points generated were analyzed by LIFEREG procedures (SAS Institute, Inc., Cary, N.C.), which showed that all parameters studied significantly affected the growth responses of S. aureus. Differences were observed in the growth-no-growth boundary when different humectants were used to achieve the desired RH values in both the absence and the presence of potassium sorbate. Sucrose plus fructose was most inhibitory at neutral pH values, while NaCl was most inhibitory at low pH values. The addition of potassium sorbate greatly increased the no-growth regions, particularly when pH was <6.0. Published kinetic growth and survival models were compared with boundary models developed in this work. The effects of solutes and differences in modeling approaches are discussed. PMID- 11916707 TI - Effect of growth conditions and staining procedure upon the subsurface transport and attachment behaviors of a groundwater protist. AB - The transport and attachment behaviors of Spumella guttula (Kent), a nanoflagellate (protist) found in contaminated and uncontaminated aquifer sediments in Cape Cod, Mass., were assessed in flowthrough and static columns and in a field injection-and-recovery transport experiment involving an array of multilevel samplers. Transport of S. guttula harvested from low-nutrient (10 mg of dissolved organic carbon per liter), slightly acidic, granular (porous) growth media was compared to earlier observations involving nanoflagellates grown in a traditional high-nutrient liquid broth. In contrast to the highly retarded (retardation factor of approximately 3) subsurface transport previously reported for S. guttula, the peak concentration of porous-medium-grown S. guttula traveled concomitantly with that of a conservative (bromide) tracer. About one-third of the porous-medium-grown nanoflagellates added to the aquifer were transported at least 2.8 m downgradient, compared to only approximately 2% of the broth-grown nanoflagellates. Flowthrough column studies revealed that a vital (hydroethidine [HE]) staining procedure resulted in considerably less attachment (more transport) of S. guttula in aquifer sediments than did a staining-and-fixation procedure involving 4',6'-diamidino-2-phenylindole (DAPI) and glutaraldehyde. The calculated collision efficiency (approximately 10(-2) for porous-medium-grown, DAPI-stained nanoflagellates) was comparable to that observed earlier for the indigenous community of unattached groundwater bacteria that serve as prey. The attachment of HE-labeled S. guttula onto aquifer sediment grains was independent of pH (over the range from pH 3 to 9) suggesting a primary attachment mechanism that may be fundamentally different from that of their prey bacteria, which exhibit sharp decreases in fractional attachment with increasing pH. The high degree of mobility of S. guttula in the aquifer sediments has important ecological implications for the protistan community within the temporally changing plume of organic contaminants in the Cape Cod aquifer. PMID- 11916708 TI - Diversity, dynamics, and activity of bacterial communities during production of an artisanal Sicilian cheese as evaluated by 16S rRNA analysis. AB - The diversity and dynamics of the microbial communities during the manufacturing of Ragusano cheese, an artisanal cheese produced in Sicily (Italy), were investigated by a combination of classical and culture-independent approaches. The latter included PCR, reverse transcriptase-PCR (RT-PCR), and denaturing gradient gel electrophoresis (DGGE) of 16S rRNA genes (rDNA). Bacterial and Lactobacillus group-specific primers were used to amplify the V6 to V8 and V1 to V3 regions of the 16S rRNA gene, respectively. DGGE profiles from samples taken during cheese production indicated dramatic shifts in the microbial community structure. Cloning and sequencing of rDNA amplicons revealed that mesophilic lactic acid bacteria (LAB), including species of Leuconostoc, Lactococcus lactis, and Macrococcus caseolyticus were dominant in the raw milk, while Streptococcus thermophilus prevailed during lactic fermentation. Other thermophilic LAB, especially Lactobacillus delbrueckii and Lactobacillus fermentum, also flourished during ripening. Comparison of the rRNA-derived patterns obtained by RT-PCR to the rDNA DGGE patterns indicated a substantially different degree of metabolic activity for the microbial groups detected. Identification of cultivated LAB isolates by phenotypic characterization and 16S rDNA analysis indicated a variety of species, reflecting to a large extent the results obtained from the 16S rDNA clone libraries, with the significant exception of the Lactobacillus delbrueckii species, which dominated in the ripening cheese but was not detected by cultivation. The present molecular approaches combined with culture can effectively describe the complex ecosystem of natural fermented dairy products, giving useful information for starter culture design and preservation of artisanal fermented food technology. PMID- 11916709 TI - Diversity of nitrite reductase (nirK and nirS) gene fragments in forested upland and wetland soils. AB - The genetic heterogeneity of nitrite reductase gene (nirK and nirS) fragments from denitrifying prokaryotes in forested upland and marsh soil was investigated using molecular methods. nirK gene fragments could be amplified from both soils, whereas nirS gene fragments could be amplified only from the marsh soil. PCR products were cloned and screened by restriction fragment length polymorphism (RFLP), and representative fragments were sequenced. The diversity of nirK clones was lower than the diversity of nirS clones. Among the 54 distinct nirK RFLP patterns identified in the two soils, only one pattern was found in both soils and in each soil two dominant groups comprised >35% of all clones. No dominance and few redundant patterns were seen among the nirS clones. Phylogenetic analysis of deduced amino acids grouped the nirK sequences into five major clusters, with one cluster encompassing most marsh clones and all upland clones. Only a few of the nirK clone sequences branched with those of known denitrifying bacteria. The nirS clones formed two major clusters with several subclusters, but all nirS clones showed less than 80% identity to nirS sequences from known denitrifying bacteria. Overall, the data indicated that the denitrifying communities in the two soils have many members and that the soils have a high richness of different nir genes, especially of the nirS gene, most of which have not yet been found in cultivated denitrifiers. PMID- 11916710 TI - Modeling yeast spoilage in cold-filled ready-to-drink beverages with Saccharomyces cerevisiae, Zygosaccharomyces bailii, and Candida lipolytica. AB - Mathematical models were developed to predict the probability of yeast spoilage of cold-filled ready-to-drink beverages as a function of beverage formulation. A Box-Behnken experimental design included five variables, each at three levels: pH (2.8, 3.3, and 3.8), titratable acidity (0.20, 0.40, and 0.60%), sugar content (8.0, 12.0, and 16.0 degrees Brix), sodium benzoate concentration (100, 225, and 350 ppm), and potassium sorbate concentration (100, 225, and 350 ppm). Duplicate samples were inoculated with a yeast cocktail (100 microl/50 ml) consisting of equal proportions of Saccharomyces cerevisiae, Zygosaccharomyces bailii, and Candida lipolytica (approximately 5.0 x 10(4) CFU/ml each). The inoculated samples were plated on malt extract agar after 0, 1, 2, 4, 6, and 8 weeks. Logistic regression was used to create the predictive models. The pH and sodium benzoate and potassium sorbate concentrations were found to be significant factors controlling the probability of yeast growth. Interaction terms for pH and each preservative were also significant in the predictive model. Neither the titratable acidity nor the sugar content of the model beverages was a significant predictor of yeast growth in the ranges tested. PMID- 11916711 TI - Engineering of phytase for improved activity at low pH. AB - For industrial applications in animal feed, a phytase of interest must be optimally active in the pH range prevalent in the digestive tract. Therefore, the present investigation describes approaches to rationally engineer the pH activity profiles of Aspergillus fumigatus and consensus phytases. Decreasing the negative surface charge of the A. fumigatus Q27L phytase mutant by glycinamidylation of the surface carboxy groups (of Asp and Glu residues) lowered the pH optimum by ca. 0.5 unit but also resulted in 70 to 75% inactivation of the enzyme. Alternatively, detailed inspection of amino acid sequence alignments and of experimentally determined or homology modeled three-dimensional structures led to the identification of active-site amino acids that were considered to correlate with the activity maxima at low pH of A. niger NRRL 3135 phytase, A. niger pH 2.5 acid phosphatase, and Peniophora lycii phytase. Site-directed mutagenesis confirmed that, in A. fumigatus wild-type phytase, replacement of Gly-277 and Tyr 282 with the corresponding residues of A. niger phytase (Lys and His, respectively) gives rise to a second pH optimum at 2.8 to 3.4. In addition, the K68A single mutation (in both A. fumigatus and consensus phytase backbones), as well as the S140Y D141G double mutation (in A. fumigatus phytase backbones), decreased the pH optima with phytic acid as substrate by 0.5 to 1.0 unit, with either no change or even a slight increase in maximum specific activity. These findings significantly extend our tools for rationally designing an optimal phytase for a given purpose. PMID- 11916712 TI - Physiological function of alcohol dehydrogenases and long-chain (C(30)) fatty acids in alcohol tolerance of Thermoanaerobacter ethanolicus. AB - A mutant strain (39E H8) of Thermoanaerobacter ethanolicus that displayed high (8% [vol/vol]) ethanol tolerance for growth was developed and characterized in comparison to the wild-type strain (39E), which lacks alcohol tolerance (<1.5% [vol/vol]). The mutant strain, unlike the wild type, lacked primary alcohol dehydrogenase and was able to increase the percentage of transmembrane fatty acids (i.e., long-chain C(30) fatty acids) in response to increasing levels of ethanol. The data support the hypothesis that primary alcohol dehydrogenase functions primarily in ethanol consumption, whereas secondary alcohol dehydrogenase functions in ethanol production. These results suggest that improved thermophilic ethanol fermentations at high alcohol levels can be developed by altering both cell membrane composition (e.g., increasing transmembrane fatty acids) and the metabolic machinery (e.g., altering primary alcohol dehydrogenase and lactate dehydrogenase activities). PMID- 11916713 TI - Towards growth of arbuscular mycorrhizal fungi independent of a plant host. AB - When surface-sterilized spores of the arbuscular mycorrhizal fungus (AMF) Glomus intraradices Sy167 were germinated on agar plates in the slightly modified minimum mineral medium described by G. Becard and J. A. Fortin (New Phytol. 108:211-218, 1988), slime-forming bacteria, identified as Paenibacillus validus, frequently grew up. These bacteria were able to support growth of the fungus on the agar plates. In the presence of P. validus, hyphae branched profusely and formed coiled structures. These were much more densely packed than the so-called arbuscule-like structures which are formed by AMF grown in coculture with carrot roots transformed with T-DNA from Agrobacterium rhizogenes. The presence of P. validus alone also enabled G. intraradices to form new spores, mainly at the densely packed hyphal coils. The new spores were not as abundant as and were smaller than those formed by AMF in the monoxenic culture with carrot root tissues, but they also contained lipid droplets and a large number of nuclei. In these experiments P. validus could not be replaced by bacteria such as Escherichia coli K-12 or Azospirillum brasilense Sp7. Although no conditions under which the daughter spores regerminate and colonize plants have been found yet, and no factor(s) from P. validus which stimulates fungal growth has been identified, the present findings might be a significant step forward toward growth of AMF independent of any plant host. PMID- 11916714 TI - Norwalk-like virus sequences in mineral waters: one-year monitoring of three brands. AB - In a recent study, RNA with nucleotide sequeces specific for "Norwalk-like viruses" (NLV) was detected in 11 different brands of European mineral waters. To clarify this finding, a 1-year monitoring study was conducted. Samples of three European brands of mineral water without gas were monitored weekly by reverse transcriptase PCR using generic and genogroup-specific oligonucleotides. Additional analyses were performed to investigate a possible correlation between NLV sequence contamination and mineral water lot numbers, the long-term stability (persistence) of NLV sequences in mineral water, and the level of contamination. NLV sequences were detected in 53 of 159 samples analyzed (33%) and belonged entirely to genogroup II. Although all NLV strains identified were closely related, three mineral water brand-specific clusters could be identified for both primer systems by sequencing. Analyses of second samples from lots previously shown to be positive for NLV sequences gave corresponding results in 45 of 53 cases (85%) (within a six-pack). NLV persistence was tested by analyzing 10 positive samples after 6 and 12 months of storage in darkness at room temperature. After 6 months, all samples remained positive; after 12 months, 9 of 10 samples were still positive for NLV sequences. No NLV sequences could be detected by analysis of 0.1-liter aliquots of 53 samples shown to be positive by testing of 1-liter volumes. Based on this fact and a test sensitivity of approximately 10 viral units, levels of contamination in positive mineral water samples were estimated to be in the range of 10 to 100 genomic equivalents per liter. PMID- 11916715 TI - Desulfovibrio sp. genes involved in the respiration of sulfate during metabolism of hydrogen and lactate. AB - To develop a better understanding of respiration by sulfate-reducing bacteria, we examined transcriptional control of respiratory genes during growth with lactate or hydrogen as an electron donor. RNA extracts of Desulfovibrio desulfuricans subsp. aestuarii were analyzed by using random arbitrarily primed PCR. RNA was reverse transcribed under low-stringency conditions with a set of random primers, and candidate cDNAs were cloned, sequenced, and characterized by BLAST analysis. Putative differentially expressed transcripts were confirmed by Northern blot analysis. Interestingly, dissimilatory bisulfite reductase was upregulated in the presence of hydrogen. To link these transcriptional changes to the physiology of sulfate-reducing bacteria, sulfide was measured during growth of several strains of Desulfovibrio on hydrogen or lactate, and this revealed that hydrogen-grown cells produced more sulfide per unit of cell mass than lactate-grown cells. Transcription of other redox proteins was characterized by Northern blotting to determine whether or not they were also transcribed to higher levels in hydrogen grown cells. Growth on lactate produced greater transcription of [NiFe] hydrogenase. H(2)-grown cells transcribed the adenylylsulfate reductase b subunit and HmcA to higher levels. The results we describe here provide new insight into the continuing debate over how Desulfovibrio species utilize redox components to generate membrane potential and to channel electrons to sulfate, the final electron acceptor. PMID- 11916716 TI - Spatially oscillating activity and microbial succession of mercury-reducing biofilms in a technical-scale bioremediation system. AB - Mercury-contaminated chemical wastewater of a mercury cell chloralkali plant was cleaned on site by a technical-scale bioremediation system. Microbial mercury reduction of soluble Hg(II) to precipitating Hg(0) decreased the mercury load of the wastewater during its flow through the bioremediation system by up to 99%. The system consisted of a packed-bed bioreactor, where most of the wastewater's mercury load was retained, and an activated carbon filter, where residual mercury was removed from the bioreactor effluent by both physical adsorption and biological reduction. In response to the oscillation of the mercury concentration in the bioreactor inflow, the zone of maximum mercury reduction oscillated regularly between the lower and the upper bioreactor horizons or the carbon filter. At low mercury concentrations, maximum mercury reduction occurred near the inflow at the bottom of the bioreactor. At high concentrations, the zone of maximum activity moved to the upper horizons. The composition of the bioreactor and carbon filter biofilms was investigated by 16S-23S ribosomal DNA intergenic spacer polymorphism analysis. Analysis of spatial biofilm variation showed an increasing microbial diversity along a gradient of decreasing mercury concentrations. Temporal analysis of the bioreactor community revealed a stable abundance of two prevalent strains and a succession of several invading mercury resistant strains which was driven by the selection pressure of high mercury concentrations. In the activated carbon filter, a lower selection pressure permitted a steady increase in diversity during 240 days of operation and the establishment of one mercury-sensitive invader. PMID- 11916717 TI - Shedding of Escherichia coli O157:H7 in dairy cattle housed in a confined environment following waterborne inoculation. AB - A study of Escherichia coli O157:H7 transmission and shedding was conducted with bull calves housed in individual pens within a confined environment. For comparative purposes, the numbers and duration of E. coli O157:H7 shedding in naturally infected calves were monitored after a single purchased calf (calf 156) tested positive prior to inoculation. During the next 8 days, the calves in adjacent pens and a pen directly across a walkway from calf 156 began to shed this serotype O157:H7 strain. Five of the eight calves in this room shed this O157:H7 strain at some time during the following 8 weeks. The numbers of E. coli O157:H7 isolates shed in these calves varied from 60 to 10(5) CFU/g of feces, and the duration of shedding ranged from 17 to >31 days. The genomic DNAs from isolates recovered from these calves were indistinguishable when compared by using XbaI digestion and pulsed-field gel electrophoresis. Inoculation of calves with 1 liter of water containing ca. 10(3) to 10(4) CFU of E. coli O157:H7/ml resulted in shedding in 10 of 12 calves (trial 1, 4 of 4 calves; trial 2, 6 of 8 calves). The inoculated calves shed the inoculation strain (FRIK 1275) as early as 24 h after administration. The duration of shedding varied from 18 to >43 days at levels from 10(2) to 10(6) CFU/g of feces. The numbers of doses necessary to initiate shedding varied among calves, and two calves in trial 2 never shed FRIK 1275 after four doses (ca. 10(6) CFU per dose). Results from this study confirm previous reports of animal-to-animal and waterborne dissemination of E. coli O157:H7 and highlight the need for an effective water treatment to reduce the spread of this pathogen in cattle. PMID- 11916718 TI - Yeast species associated with orange juice: evaluation of different identification methods. AB - Five different methods were used to identify yeast isolates from a variety of citrus juice sources. A total of 99 strains, including reference strains, were identified using a partial sequence of the 26S rRNA gene, restriction pattern analysis of the internal transcribed spacer region (5.8S-ITS), classical methodology, the RapID Yeast Plus system, and API 20C AUX. Twenty-three different species were identified representing 11 different genera. Distribution of the species was considerably different depending on the type of sample. Fourteen different species were identified from pasteurized single-strength orange juice that had been contaminated after pasteurization (PSOJ), while only six species were isolated from fresh-squeezed, unpasteurized orange juice (FSOJ). Among PSOJ isolates, Candida intermedia and Candida parapsilosis were the predominant species. Hanseniaspora occidentalis and Hanseniaspora uvarum represented up to 73% of total FSOJ isolates. Partial sequence of the 26S rRNA gene yielded the best results in terms of correct identification, followed by classical techniques and 5.8S-ITS analysis. The commercial identification kits RapID Yeast Plus system and API 20C AUX were able to correctly identify only 35 and 13% of the isolates, respectively. Six new 5.8S-ITS profiles were described, corresponding to Clavispora lusitaniae, Geotrichum citri-aurantii, H. occidentalis, H. vineae, Pichia fermentans, and Saccharomycopsis crataegensis. With the addition of these new profiles to the existing database, the use of 5.8S-ITS sequence became the best tool for rapid and accurate identification of yeast isolates from orange juice. PMID- 11916719 TI - Development and characterization of a green fluorescent protein-based bacterial biosensor for bioavailable toluene and related compounds. AB - A green fluorescent protein-based Pseudomonas fluorescens strain A506 biosensor was constructed and characterized for its potential to measure benzene, toluene, ethylbenzene, and related compounds in aqueous solutions. The biosensor is based on a plasmid carrying the toluene-benzene utilization (tbu) pathway transcriptional activator TbuT from Ralstonia pickettii PKO1 and a transcriptional fusion of its promoter PtbuA1 with a promoterless gfp gene on a broad-host-range promoter probe vector. TbuT was not limiting, since it was constitutively expressed by being fused to the neomycin phosphotransferase (nptII) promoter. The biosensor cells were readily induced, and fluorescence emission after induction periods of 3 h correlated well with toluene, benzene, ethylbenzene, and trichloroethylene concentrations. Our experiments using flow cytometry show that intermediate levels of gfp expression in response to toluene reflect uniform induction of cells. As the toluene concentration increases, the level of gfp expression per cell increases until saturation kinetics of the TbuT PtbuA1 system are observed. Each inducer had a unique minimum concentration that was necessary for induction, with K(app) values that ranged from 3.3 +/- 1.8 microM for toluene to 35.6 +/- 16.6 microM for trichloroethylene (means +/- standard errors of the means), and maximal fluorescence response. The fluorescence response was specific for alkyl-substituted benzene derivatives and branched alkenes (di- and trichloroethylene, 2-methyl-2-butene). The biosensor responded in an additive fashion to the presence of multiple inducers and was unaffected by the presence of compounds that were not inducers, such as those present in gasoline. Flow cytometry revealed that, in response to toxic concentrations of gasoline, there was a small uninduced population and another larger fully induced population whose levels of fluorescence corresponded to the amount of effectors present in the sample. These results demonstrate the potential for green fluorescent protein-based bacterial biosensors to measure environmental contaminants. PMID- 11916720 TI - Expanded genetic map of Gibberella moniliformis (Fusarium verticillioides). AB - Gibberella moniliformis (Fusarium verticillioides) is primarily a pathogen of maize, but it can also cause disease in other crop species. This pathogenicity, as well as the contamination of food- and feedstuffs with the fumonisin mycotoxins, results in economically significant losses to both farmers and food processors. The dissection of important biological characters in this fungus has been hampered by the lack of a uniformly dense genetic map. The existing restriction fragment length polymorphism-based map contains significant gaps, making it difficult to routinely locate biologically important genes, such as those involved in pathogenicity or mycotoxin production, with precision. We utilized amplified fragment length polymorphisms (AFLPs) to saturate the existing genetic map and added 486 AFLP markers to the approximately 150 markers on the existing map. The resulting map has an average marker interval of 3.9 map units and averages approximately 21 kb/map unit. The additional markers expanded the map from 1,452 to 2,188 map units distributed across 12 chromosomes. The maximum distance between adjacent markers is 29 map units. We identified AFLP markers less than 1 map unit from the mating type (MAT) locus and 2.5 map units from the spore killer (SK) locus; eight AFLP markers map within 8.5 units of the FUM1 (fumonisin biosynthetic) locus. The increased saturation of this map will facilitate further development of G. moniliformis as a model system for the genetic and population genetic studies of related, but less genetically tractable, plant pathogenic fungi. PMID- 11916721 TI - Purification and characterization of an arginine aminopeptidase from Lactobacillus sakei. AB - An arginine aminopeptidase (EC 3.4.11.6) that exclusively hydrolyzes basic amino acids from the amino (N) termini of peptide substrates has been purified from Lactobacillus sakei. The purification procedure consisted of ammonium sulfate fractionation and three chromatographic steps, which included hydrophobic interaction, gel filtration, and anion-exchange chromatography. This procedure resulted in a recovery rate of 4.2% and a 500-fold increase in specific activity. The aminopeptidase appeared to be a trimeric enzyme with a molecular mass of 180 kDa. The activity was optimal at pH 5.0 and 37 degrees C. The enzyme was inhibited by sulfhydryl group reagents and several divalent cations (Cu(2+), Hg(2+), and Zn(2+)) but was activated by reducing agents, metal-chelating agents, and sodium chloride. The enzyme showed a preference for arginine at the N termini of aminoacyl derivatives and peptides. The K(m) values for Arg-7-amido-4 methylcoumarin (AMC) and Lys-AMC were 15.9 and 26.0 microM, respectively. The nature of the amino acid residue at the C terminus of dipeptides has an effect on hydrolysis rates. The activity was maximal toward dipeptides with Arg, Lys, or Ala as the C-terminal residue. The properties of the purified enzyme, its potential function in the release of arginine, and its further metabolism are discussed because, as a whole, it could constitute a survival mechanism for L. sakei in the meat environment. PMID- 11916722 TI - Inducible gene expression by nonculturable bacteria in milk after pasteurization. AB - The viability of bacteria in milk after heat treatments was assessed by using three different viability indicators: (i) CFU on plate count agar, (ii) de novo expression of a gfp reporter gene, and (iii) membrane integrity based on propidium iodide exclusion. In commercially available pasteurized milk, direct viable counts, based on dye exclusion, were significantly (P < 0.05) higher than viable cell counts determined from CFU, suggesting that a significant subpopulation of cells in pasteurized milk are viable but nonculturable. Heating milk at 63.5 degrees C for 30 min resulted in a >4-log-unit reduction in the number of CFU of Escherichia coli and Pseudomonas putida that were marked with lac-inducible gfp. However, the reduction in the number of gfp-expressing cells of both organisms under the same conditions was <2.5 log units. These results demonstrate that a substantial portion of cells rendered incapable of forming colonies by heat treatment are metabolically active and are able to transcribe and translate genes de novo. PMID- 11916723 TI - Microbial diversity of hydrothermal sediments in the Guaymas Basin: evidence for anaerobic methanotrophic communities. AB - Microbial communities in hydrothermally active sediments of the Guaymas Basin (Gulf of California, Mexico) were studied by using 16S rRNA sequencing and carbon isotopic analysis of archaeal and bacterial lipids. The Guaymas sediments harbored uncultured euryarchaeota of two distinct phylogenetic lineages within the anaerobic methane oxidation 1 (ANME-1) group, ANME-1a and ANME-1b, and of the ANME-2c lineage within the Methanosarcinales, both previously assigned to the methanotrophic archaea. The archaeal lipids in the Guaymas Basin sediments included archaeol, diagnostic for nonthermophilic euryarchaeota, and sn-2 hydroxyarchaeol, with the latter compound being particularly abundant in cultured members of the Methanosarcinales. The concentrations of these compounds were among the highest observed so far in studies of methane seep environments. The delta-(13)C values of these lipids (delta-(13)C = -89 to -58 per thousand) indicate an origin from anaerobic methanotrophic archaea. This molecular-isotopic signature was found not only in samples that yielded predominantly ANME-2 clones but also in samples that yielded exclusively ANME-1 clones. ANME-1 archaea therefore remain strong candidates for mediation of the anaerobic oxidation of methane. Based on 16S rRNA data, the Guaymas sediments harbor phylogenetically diverse bacterial populations, which show considerable overlap with bacterial populations of geothermal habitats and natural or anthropogenic hydrocarbon-rich sites. Consistent with earlier observations, our combined evidence from bacterial phylogeny and molecular-isotopic data indicates an important role of some novel deeply branching bacteria in anaerobic methanotrophy. Anaerobic methane oxidation likely represents a significant and widely occurring process in the trophic ecology of methane-rich hydrothermal vents. This study stresses a high diversity among communities capable of anaerobic oxidation of methane. PMID- 11916724 TI - Statistical analysis of Pseudomonas aeruginosa biofilm development: impact of mutations in genes involved in twitching motility, cell-to-cell signaling, and stationary-phase sigma factor expression. AB - Four strains of Pseudomonas aeruginosa (wild type, Delta(pil)HIJK mutant, lasI mutant, and rpoS mutant) were genetically tagged with the green fluorescent protein, and the development of flow chamber-grown biofilms by each of them was investigated by confocal laser scanning microscopy. The structural developments of the biofilms were quantified by the computer program COMSTAT (A. Heydorn, A. T. Nielsen, M. Hentzer, C. Sternberg, M. Givskov, B. K. Ersboll, and S. Molin, Microbiology 146:2395-2407, 2000). Two structural key variables, average thickness and roughness, formed the basis for an analysis of variance model comprising the four P. aeruginosa strains, five time points (55, 98, 146, 242, and 314 h), and three independent rounds of biofilm experiments. The results showed that the wild type, the Delta(pil)HIJK mutant, and the rpoS mutant display conspicuously different types of temporal biofilm development, whereas the lasI mutant was indistinguishable from the wild type at all time points. The wild type and the lasI mutant formed uniform, densely packed biofilms. The rpoS mutant formed densely packed biofilms that were significantly thicker than those of the wild type, whereas the Delta(pil)HIJK mutant formed distinct microcolonies that were regularly spaced and almost uniform in size. The results are discussed in relation to the current model of P. aeruginosa biofilm development. PMID- 11916725 TI - The Awa1 gene is required for the foam-forming phenotype and cell surface hydrophobicity of sake yeast. AB - Sake, a traditional alcoholic beverage in Japan, is brewed with sake yeasts, which are classified as Saccharomyces cerevisiae. Almost all sake yeasts form a thick foam layer on sake mash during the fermentation process because of their cell surface hydrophobicity, which increases the cells' affinity for bubbles. To reduce the amount of foam, nonfoaming mutants were bred from foaming sake yeasts. Nonfoaming mutants have hydrophilic cell surfaces and no affinity for bubbles. We have cloned a gene from a foam-forming sake yeast that confers foaming ability to a nonfoaming mutant. This gene was named AWA1 and structures of the gene and its product were analyzed. The N- and C-terminal regions of Awa1p have the characteristic sequences of a glycosylphosphatidylinositol anchor protein. The entire protein is rich in serine and threonine residues and has a lot of repetitive sequences. These results suggest that Awa1p is localized in the cell wall. This was confirmed by immunofluorescence microscopy and Western blotting analysis using hemagglutinin-tagged Awa1p. Moreover, an awa1 disruptant of sake yeast was hydrophilic and showed a nonfoaming phenotype in sake mash. We conclude that Awa1p is a cell wall protein and is required for the foam-forming phenotype and the cell surface hydrophobicity of sake yeast. PMID- 11916726 TI - Bacterial cell surface display of organophosphorus hydrolase for selective screening of improved hydrolysis of organophosphate nerve agents. AB - Organophosphorus hydrolase (OPH) is a bacterial enzyme that has been shown to degrade a wide range of neurotoxic organophosphate nerve agents. However, the effectiveness of degradation varies dramatically, ranging from highly efficient with paraoxon to relatively slow with methyl parathion. Sequential cycles of DNA shuffling and screening were used to fine-tune and enhance the activity of OPH towards poorly degraded substrates. Because of the inaccessibility of these pesticides across the cell membrane, OPH variants were displayed on the surface of Escherichia coli using the truncated ice nucleation protein in order to isolate novel enzymes with truly improved substrate specificities. A solid-phase top agar method based on the detection of the yellow product p-nitrophenol was developed for the rapid prescreening of potential variants with improved hydrolysis of methyl parathion. Two rounds of DNA shuffling and screening were carried out, and several improved variants were isolated. One variant in particular, 22A11, hydrolyzes methyl parathion 25-fold faster than does the wild type. Because of the success that we achieved with directed evolution of OPH for improved hydrolysis of methyl parathion, we believe that we can easily extend this method in creating other OPH variants with improved activity against poorly degraded pesticides such as diazinon and chlorpyrifos and nerve agents such as sarin and soman. PMID- 11916727 TI - Rapid assessment of the physiological status of the polychlorinated biphenyl degrader Comamonas testosteroni TK102 by flow cytometry. AB - The viability of the polychlorinated biphenyl-degrading bacterium Comamonas testosteroni TK102 was assessed by flow cytometry (FCM) with the fluorogenic ester Calcein-AM (CAM) and the nucleic acid dye propidium iodide (PI). CAM stained live cells, whereas PI stained dead cells. When double staining with CAM and PI was performed, three physiological states, i.e., live (calcein positive, PI negative), dead (calcein negative, PI positive), and permeabilized (calcein positive, PI positive), were detected. To evaluate the reliability of this double staining method, suspensions of live and dead cells were mixed in various proportions and analyzed by FCM. The proportion of dead cells measured by FCM directly correlated with the proportion of dead cells in the sample (y = 0.9872 x + 0.18; R(2) = 0.9971). In addition, the proportion of live cells measured by FCM inversely correlated with the proportion of dead cells in the sample (y = -0.9776 x + 98.36; R(2) = 0.9962). The proportion of permeabilized cells was consistently less than 2%. These results indicate that FCM in combination with CAM and PI staining is rapid (Val) [DNA: C-to-T substitution at nucleo-tide 677 (677C-->T)] in methylenetetrahydrofolate reductase (NADPH) and the cofactor FAD (in relation to cardiovascular disease, migraines, and rages), the Pro187-->Ser (DNA: 609C-->T) mutation in NAD(P):quinone oxidoreductase 1 [NAD(P)H dehy-drogenase (quinone)] and FAD (in relation to cancer), the Ala44-->Gly (DNA: 131C-->G) mutation in glucose-6-phosphate 1-dehydrogenase and NADP (in relation to favism and hemolytic anemia), and the Glu487-->Lys mutation (present in one-half of Asians) in aldehyde dehydrogenase (NAD + ) and NAD (in relation to alcohol intolerance, Alzheimer disease, and cancer). PMID- 11916750 TI - Collateral damage in the battle against hypovitaminosis A? PMID- 11916751 TI - The dopamine mesocorticolimbic pathway is affected by deficiency in n-3 polyunsaturated fatty acids. AB - BACKGROUND: Several findings in humans support the hypothesis of links between n 3 polyunsaturated fatty acid (PUFA) status and psychiatric diseases. OBJECTIVE: The involvement of PUFAs in central nervous system function can be assessed with the use of dietary manipulation in animal models. We studied the effects of chronic dietary n-3 PUFA deficiency on mesocorticolimbic dopamine neurotransmission in rats. DESIGN: Using dual-probe microdialysis, we analyzed dopamine release under amphetamine stimulation simultaneously in the frontal cortex and the nucleus accumbens. The messenger RNA (mRNA) expression of vesicular monoamine transporter(2) and dopamine D(2) receptor was studied with the use of in situ hybridization. The protein expression of the synthesis limiting enzyme tyrosine 3-monooxygenase (tyrosine 3-hydroxylase) was studied with the use of immunocytochemistry. RESULTS: Dopamine release was significantly lower in both cerebral areas in n-3 PUFA-deficient rats than in control rats, but this effect was abolished in the frontal cortex and reversed in the nucleus accumbens by reserpine pretreatment, which depletes the dopamine vesicular storage pool. The mRNA expression of vesicular monoamine transporter(2) was lower in both cerebral areas in n-3 PUFA-deficient rats than in control rats, whereas the mRNA expression of D(2) receptor was lower in the frontal cortex and higher in the nucleus accumbens in n-3 PUFA-deficient rats than in control rats. Finally, tyrosine 3-monooxygenase immunoreactivity was higher in the ventral tegmental area in n-3 PUFA-deficient rats than in control rats. CONCLUSIONS: Our results suggest that the mesolimbic dopamine pathway is more active whereas the mesocortical pathway is less active in n-3 PUFA-deficient rats than in control rats. This provides new neurochemical evidence supporting the effects of n-3 PUFA deficiency on behavior. PMID- 11916752 TI - Increased plasma fatty acid concentrations after respiratory exacerbations are associated with elevated oxidative stress in cystic fibrosis patients. AB - BACKGROUND: Oxidative stress and depleted antioxidant defenses occur in stable cystic fibrosis patients. During acute infection, the balance between oxidants and antioxidants may be further disturbed. OBJECTIVE: We examined the oxidative stress during acute infection in cystic fibrosis patients by measuring 8-iso prostaglandin F(2 alpha) (8-iso-PGF(2 alpha)) and antioxidant defenses in relation to dietary intake, fatty acid status, immune function, and clinical status. DESIGN: Plasma concentrations of total 8-iso-PGF(2 alpha), vitamins E and C, beta-carotene, zinc, selenium, and copper; plasma fatty acid compositions; erythrocyte glutathione concentrations; glutathione peroxidase and superoxide dismutase activity; sputum glutathione and 8-iso-PGF(2 alpha) concentrations; lung function; clinical symptoms; and dietary intake were measured in 15 cystic fibrosis patients before and after 10-14 d of intravenous antibiotic treatment for a pulmonary exacerbation. RESULTS: After treatment, respiratory status improved (percentage of forced expiratory volume in 1 s: 60 +/- 6% at baseline compared with 74 +/- 7% after treatment, P = 0.01), quality of well-being improved (P = 0.001), and total plasma 8-iso-PGF(2 alpha) concentrations increased from 469 nmol/L at baseline (interquartile range: 373-554 nmol/L) to 565 nmol/L after treatment (interquartile range: 429-689 nmol/L; P = 0.008). Total energy, fat, carbohydrate, and protein intakes per kilogram body weight also increased; however, dietary antioxidant intake was unchanged. Plasma fatty acid concentrations increased after treatment, strongly correlating with plasma 8 iso-PGF(2 alpha) concentrations (r = 0.768, P = 0.001). There were no significant changes in white cell counts or plasma concentrations of vitamins E and C or beta carotene. Erythrocyte glutathione peroxidase activity was reduced after treatment, whereas there was no significant change in superoxide dismutase activity. CONCLUSIONS: Oxidative stress increased after treatment for pulmonary exacerbations and was strongly linked to increased concentrations of plasma fatty acids. Although intravenous antibiotic therapy and physiotherapy improved lung function within 10-14 d of treatment, the biochemical effects of oxidation continued further. Thus, antioxidant intervention during treatment for and recovery from acute infection in cystic fibrosis should be considered. PMID- 11916753 TI - Birth weight and body composition in young women: a prospective twin study. AB - BACKGROUND: The intrauterine environment may be critical for the development of obesity. Alternatively, the same genetic factors may influence both birth weight and adult body composition. OBJECTIVE: We evaluated the association between birth weight and adult body composition in female twins, which allowed us to control for maternal and genetic influences. DESIGN: Of 447 twin pairs randomly selected from the East Flanders Prospective Twin Survey, 238 pairs, aged 18-34 y, participated. Adult body mass, height, and lean body mass were measured, and the body mass index (BMI), waist-to-hip ratio, and sum of skinfold thicknesses were calculated. The twins were considered as individuals and pairs. RESULTS: When the twins were considered as individuals, twins who were heavier at birth were taller (3.3 cm/kg greater birth weight) and slightly heavier (1.13 kg/kg greater birth weight) as adults than were lighter twins. They also had more lean body mass and less subcutaneous and abdominal fat at birth. Pairwise comparison showed that for every level of intrapair birth weight difference (> or = 5%, > or = 10%, and > or = 15%), the twin who was heavier at birth was taller in adult life (0.8, 1.2, and 2.0 cm, respectively). When the intrapair birth weight difference exceeded 15%, the heavier twin was also heavier (3.1 +/- 6.08 kg) as an adult than her much lighter sister. CONCLUSION: Birth weight accounts for some of the differences in adult body composition between twins. PMID- 11916754 TI - Body mass index and waist circumference independently contribute to the prediction of nonabdominal, abdominal subcutaneous, and visceral fat. AB - BACKGROUND: It is unknown whether the ability of waist circumference (WC) to predict health risk beyond that predicted by body mass index (BMI) alone is explained in part by the ability of WC to identify those with elevated concentrations of total or abdominal fat. OBJECTIVE: We sought to determine whether BMI and WC independently contribute to the prediction of non-abdominal (total fat - abdominal fat), abdominal subcutaneous, and visceral fat. DESIGN: Fat distribution was measured by magnetic resonance imaging in 341 white men and women. Multiple regression analysis was performed to measure whether the combination of BMI and WC explained a greater variance in non-abdominal, abdominal subcutaneous, and visceral fat than did BMI or WC alone. These fat depots were also compared after a subdivision of the cohort into 3 BMI (normal, overweight, and class I obese) and 3 WC (low, intermediate, and high) categories according to the classification system used to identify associations between BMI, WC, and health risk. RESULTS: Independent of age and sex, the combination of BMI and WC explained a greater variance in non-abdominal, abdominal subcutaneous, and visceral fat than did either BMI or WC alone (P < 0.05). For non-abdominal and abdominal subcutaneous fat, BMI was the strongest correlate; thus, by adding BMI to WC, the variance accrued was greater than when WC was added to BMI. However, when WC was added to BMI, the added variance explained for visceral fat was greater than when BMI was added to WC. Furthermore, within each of the 3 BMI categories studied, an increase in the WC category was associated with an increase in visceral fat (P < 0.05). CONCLUSIONS: BMI and WC independently contribute to the prediction of non-abdominal, abdominal subcutaneous, and visceral fat in white men and women. These observations reinforce the importance of using both BMI and WC in clinical practice. PMID- 11916755 TI - Intakes of energy and macronutrients and the risk of benign prostatic hyperplasia. AB - BACKGROUND: Benign prostatic hyperplasia (BPH) is a common disease of older men. Although the etiology remains unclear, nutritional factors may have an effect on the disease. OBJECTIVE: Because the literature on the relations between macronutrient intakes and BPH risk is limited, we examined these relations among men in the Health Professionals Follow-up Study. DESIGN: We followed men aged 40 75 y from baseline in 1986 to 1994. Total BPH cases (n = 3523) comprised men who reported BPH surgery (n = 1589) or who did not undergo surgery but scored 15-35 points on the lower urinary tract symptom questionnaire of the American Urological Association (n = 1934); non-cases were men who scored < or = 7 points (n = 24388). Odds ratios (ORs) and 95% CIs were calculated by using multivariate logistic regression. RESULTS: The ORs rose with increasing total energy intake in a comparison of the highest and lowest quintiles for total BPH (OR: 1.29; 95% CI: 1.14, 1.45) and symptoms of BPH (1.43; 1.23, 1.66). Energy-adjusted total protein intake was positively associated with total BPH (1.18; 1.05, 1.33) and BPH surgery (1.26; 1.06, 1.49). Energy-adjusted total fat intake was not associated with risk of total BPH, but intakes of eicosapentaenoic, docosahexaenoic, and arachidonic acids were associated with a moderate increase in risk of total BPH. CONCLUSIONS: We observed modest direct associations between BPH and intakes of total energy, protein, and specific long-chain polyunsaturated fatty acids. Because eicosapentaenoic, docosahexaenoic, and arachidonic acids are highly unsaturated fatty acids, our findings support a possible role of oxidative stress in the etiology of BPH. PMID- 11916756 TI - Threonine requirement of healthy adults, derived with a 24-h indicator amino acid balance technique. AB - BACKGROUND: Because we question the validity of the 1985 FAO/ WHO/UNU upper requirement for threonine of 7 mg x kg(-1) x d(-1), we proposed a tentative mean requirement of 15 mg x kg(-1) x d(-1). OBJECTIVE: Our goal was to assess threonine adequacy at 3 test intakes and the consequences of a 6-d compared with a 13-d dietary adaptation phase. DESIGN: We used a 24-h indicator amino acid balance technique ([1-(13)C]leucine as indicator) to assess the threonine requirement. Fifteen healthy adults were randomly assigned to receive 7, 15, or 46 mg threonine x kg(-1) x d(-1) and were studied after 6 and 13 d of adaptation to the experimental diets. Diets were based on an L-amino acid mixture in which the threonine content was varied. At 1700 on days 6 and 13, a 24-h intravenous [(13)C]leucine tracer infusion protocol was begun to assess leucine oxidation and daily leucine balances. RESULTS: There was no detectable effect of duration of dietary adaptation in leucine oxidation or balance, but the 24-h leucine oxidation and balances differed significantly between the 7-mg intake and each of the 2 higher intakes (P < 0.05). The latter were not significantly different. The 24-h leucine oxidation rate decreased across threonine intakes (P < 0.01 for main effect of diet, independent of infusion day). Leucine oxidation was highly correlated (r = 0.80) between the 2 dietary adaptation phases across all test intakes. CONCLUSION: The 1985 FAO/WHO/UNU threonine recommendation is inadequate, and 15 mg x kg(-1) x d(-1) is sufficient to achieve mean indicator (leucine) amino acid balance. PMID- 11916757 TI - Energy expenditure in preadolescent African American and white boys and girls: the Baton Rouge Children's Study. AB - BACKGROUND: Low energy expenditure has been identified as a potential risk factor for body fat gain. OBJECTIVE: The objective was to determine the relations between race, sex, body fat, and energy expenditure. DESIGN: As part of the Baton Rouge Children's Study, energy expenditure was examined in 131 preadolescent African American and white girls and boys, further stratified as obese or lean. Total daily energy expenditure (TDEE) was measured by the doubly labeled water method. Resting metabolic rate (RMR) and the thermic effect of food were measured by indirect calorimetry. Fat-free mass and fat mass were measured by dual-energy X-ray absorptiometry. To account for differences in body size, energy expenditure variables were adjusted with the use of fat-free mass or fat-free mass and fat mass as covariates. RESULTS: The African American children had lower TDEE and RMR than did the white children. A lower level of energy expended in physical activity by the African American girls and a lower RMR in the African American boys accounted for the racial differences in TDEE. The white boys had a higher RMR than did the white girls. The girls had a lower TDEE and expended less energy in activity than did the boys. Energy expended in activity was lower in the obese children. CONCLUSIONS: The African American children expended less energy than did the white children. The obese children spent less time engaged in activity or engaged in lower-intensity activity. Obese children may maintain their obese state by spending less time in physical activity, but they do not have a reduced RMR or thermic effect of food. PMID- 11916758 TI - Racial differences in the relation between uncoupling protein genes and resting energy expenditure. AB - BACKGROUND: Lower resting energy expenditure (REE) in African American women may contribute to their obesity. The identification of uncoupling protein (UCP) genes has fueled a search for genes involved in energy metabolism in humans. OBJECTIVE: We examined variation in REE in relation to variation in UCP1, UCP2, and UCP3 in 141 women aged 18-21 y. DESIGN: Standard methods were used for REE measurements and genetic analysis. Body composition was determined with the use of dual-energy X-ray absorptiometry. Multivariate analysis was used to examine the effect of genotypes on REE and on fat mass in relation to other potentially confounding variables. RESULTS: REE was 295 kJ/d lower in African American women than in white women. No significant variation in REE was seen for UCP1, UCP2, and UCP3 (p 55; exon 3a; and exon 3b) variants after adjustment for other variables including smoking status. For the UCP3 exon 5 variant, REE was significantly (P = 0.019) lower in African American women with the CC genotype than in those with the TT genotype. In African American women, there was a significant trend (P = 0.012) toward lower REE and a weak but nonsignificant trend (P = 0.1) toward greater fat mass across the 3 genotypes (TT, CT, and CC). CONCLUSIONS: The significant and dose-dependent relation between lower REE and the C allele suggests that it may be a thrifty allele. The presence of this parsimonious energy metabolism in African American women, possibly linked to UCP3, may be implicated in their susceptibility to obesity. The absence of a UCP3 effect in white women is intriguing and needs to be explored to further understand possible interactions between UCP3 and other genes. PMID- 11916759 TI - A double-blind, placebo-controlled study of vitamin A and zinc supplementation in persons with tuberculosis in Indonesia: effects on clinical response and nutritional status. AB - BACKGROUND: The results of cross-sectional studies indicate that micronutrient deficiencies are common in patients with tuberculosis. No published data exist on the effect of vitamin A and zinc supplementation on antituberculosis treatment. OBJECTIVE: Our goal was to investigate whether vitamin A and zinc supplementation increases the efficacy of antituberculosis treatment with respect to clinical response and nutritional status. DESIGN: In this double-blind, placebo-controlled trial, patients with newly diagnosed tuberculosis were divided into 2 groups. One group (n = 40) received 1500 retinol equivalents (5000 IU) vitamin A (as retinyl acetate) and 15 mg Zn (as zinc sulfate) daily for 6 mo (micronutrient group). The second group (n = 40) received a placebo. Both groups received the same antituberculosis treatment recommended by the World Health Organization. Clinical examinations, assessments of micronutrient status, and anthropometric measurements were carried out before and after 2 and 6 mo of antituberculosis treatment. RESULTS: At baseline, 64% of patients had a body mass index (in kg/m(2)) < 18.5, 32% had plasma retinol concentrations < 0.70 micromol/L, and 30% had plasma zinc concentrations < 10.7 micromol/L. After antituberculosis treatment, plasma zinc concentrations were not significantly different between groups. Plasma retinol concentrations were significantly higher in the micronutrient group than in the placebo group after 6 mo (P < 0.05). Sputum conversion (P < 0.05) and resolution of X-ray lesion area (P < 0.01) occurred earlier in the micronutrient group. CONCLUSION: Vitamin A and zinc supplementation improves the effect of tuberculosis medication after 2 mo of antituberculosis treatment and results in earlier sputum smear conversion. PMID- 11916760 TI - Effects of alpha-tocopherol supplementation and continuous subcutaneous insulin infusion on oxidative stress in Korean patients with type 2 diabetes. AB - BACKGROUND: Most Koreans with type 2 diabetes are insulin deficient and insulin resistant. Continuous subcutaneous insulin infusion (CSII) provides a suitable amount of insulin to overcome insulin deficiency and achieve near-normal blood glucose concentrations. Our previous study showed, however, that CSII does not reduce oxidative stress even though it normalizes blood glucose concentrations. OBJECTIVE: The purpose of this study was to determine whether CSII plus alpha tocopherol supplementation for 2 mo would alter oxidative stress in Korean patients with type 2 diabetes. DESIGN: Ninety-eight subjects received CSII plus either 200 mg alpha-tocopherol/d (n = 48) or a placebo (n = 50) for 2 mo. The general characteristics (age, duration of diabetes, body mass index, and blood glucose concentrations) of the 2 groups were not significantly different. RESULTS: Fasting and postprandial blood glucose concentrations of all subjects were normalized after CSII. Fasting plasma insulin concentrations did not differ significantly between the 2 groups after CSII. Lipid peroxide concentrations in plasma and red blood cells decreased and alpha-tocopherol concentrations in plasma and red blood cells increased after alpha-tocopherol supplementation. However, these changes were not affected significantly by CSII. Plasma vitamin C concentrations increased significantly after CSII plus alpha-tocopherol supplementation. However, the activities of antioxidant enzymes in red blood cells did not change significantly after CSII plus alpha-tocopherol supplementation. CONCLUSION: alpha-Tocopherol supplementation was beneficial in decreasing blood lipid peroxide concentrations without altering antioxidant enzyme activities in Korean patients with type 2 diabetes treated with CSII. PMID- 11916761 TI - Marginal iron deficiency without anemia impairs aerobic adaptation among previously untrained women. AB - BACKGROUND: Iron deficiency without anemia has been shown to reduce both muscle tissue oxidative capacity and endurance in animals. However, the consequences of iron deficiency in humans remain unclear. OBJECTIVE: We investigated the effects of iron supplementation on adaptation to aerobic training among marginally iron depleted women. We hypothesized that iron supplementation for 6 wk would significantly improve iron status and maximal oxygen uptake (VO(2)max) after 4 wk of concurrent aerobic training. DESIGN: Forty-one untrained, iron-depleted, nonanemic women were randomly assigned to receive either 50 mg FeSO(4) or a placebo twice daily for 6 wk in a double-blind trial. All subjects trained on cycle ergometers 5 d/wk for 4 wk, beginning on week 3 of the study. RESULTS: Six weeks of iron supplementation significantly improved serum ferritin and serum transferrin receptor (sTfR) concentrations and transferrin saturation without affecting hemoglobin concentrations or hematocrit. Average VO(2)max and maximal respiratory exchange ratio improved in both the placebo and iron groups after training; however, the iron group experienced significantly greater improvements in VO(2)max. Both iron-status and fitness outcomes were analyzed after stratifying by baseline sTfR concentration (> and < or = 8.0 mg/L), which showed that the previously observed treatment effects were due to iron-status and fitness improvements among subjects with poor baseline iron status. CONCLUSIONS: Our findings strongly suggest that iron deficiency without anemia but with elevated sTfR status impairs aerobic adaptation among previously untrained women and that this can be corrected with iron supplementation. PMID- 11916763 TI - Green tea extract decreases muscle necrosis in mdx mice and protects against reactive oxygen species. AB - BACKGROUND: Duchenne muscular dystrophy is a severe X-linked congenital disorder characterized by lethal muscle wasting caused by the absence of the structural protein dystrophin. OBJECTIVE: Because generation of reactive oxygen species appears to play an important role in the pathogenesis of this disease, we tested whether antioxidant green tea extract could diminish muscle necrosis in the mdx mouse dystrophy model. DESIGN: A diet supplemented with 0.01% or 0.05% green tea extract was fed to dams and neonates for 4 wk beginning on the day of birth. Muscle necrosis and regeneration were determined in stained cryosections of soleus and elongator digitorum longus muscles. Radical scavenging by green tea extract was determined in differentiated cultured C2C12 cells treated with tert butylhydroperoxide, with the use of 2',7'-dichlorofluorescin diacetate as a radical detector. RESULTS: This feeding regimen significantly and dose dependently reduced necrosis in the fast-twitch muscle elongator digitorum longus but at the doses tested had no effect on the slow-twitch soleus muscle. Green tea extract concentration-dependently decreased oxidative stress induced by tert butylhydroperoxide treatment of cultured mouse C2C12 myotubes. The lower effective dose tested in mdx mice corresponds to approximately equal to 1.4 L (7 cups) green tea/d in humans. CONCLUSION: Green tea extract may improve muscle health by reducing or delaying necrosis in mdx mice by an antioxidant mechanism. PMID- 11916762 TI - Treatment of iron deficiency in goitrous children improves the efficacy of iodized salt in Cote d'Ivoire. AB - BACKGROUND: In many developing countries, children are at high risk of both goiter and iron deficiency anemia. Iron deficiency adversely affects thyroid metabolism and may reduce the efficacy of iodine prophylaxis in areas of endemic goiter. OBJECTIVE: The aim of this study was to determine whether iron supplementation in goitrous, iron-deficient children would improve their response to iodized salt. DESIGN: We conducted a randomized, double-blind, placebo controlled trial in 5-14-y-old children in Cote d'Ivoire. Goitrous, iron deficient children (n = 166) consuming iodized salt (10-30 mg I/kg salt at the household level) were supplemented with either iron (60 mg Fe/d, 4 d/wk for 16 wk) or placebo. At 0, 1, 6, 12, and 20 wk, we measured hemoglobin, serum ferritin, serum transferrin receptor, whole-blood zinc protoporphyrin, thyrotropin, thyroxine, urinary iodine, and thyroid gland volume (by ultrasonography). RESULTS: Hemoglobin and iron status at 20 wk were significantly better after iron treatment than after placebo (P < 0.05). At 20 wk, the mean reduction in thyroid size in the iron-treated group was nearly twice that in the placebo group (x +/- SD percentage change in thyroid volume from baseline: -22.8 +/- 10.7% compared with -12.7 +/- 10.1%; P < 0.01). At 20 wk, goiter prevalence was 43% in the iron-treated group compared with 62% in the placebo group (P < 0.02). There were no significant differences between groups in whole-blood thyrotropin or serum thyroxine at baseline or during the intervention. CONCLUSIONS: Iron supplementation improves the efficacy of iodized salt in goitrous children with iron deficiency. A high prevalence of iron deficiency among children in areas of endemic goiter may reduce the effectiveness of iodine prophylaxis. PMID- 11916765 TI - Weight-for-stature compared with body mass index-for-age growth charts for the United States from the Centers for Disease Control and Prevention. AB - BACKGROUND: The 2000 Centers for Disease Control and Prevention growth charts for the United States include population reference data for body mass index (BMI)-for age (ages 2-19 y) and weight-for-stature (from 77 to 121 cm). For younger children, either set of reference data could be used. OBJECTIVE: The objective of this study was to compare BMI-for-age with weight-for-stature. DESIGN: We used data for 4348 children (aged 2-5 y) from the third National Health and Nutrition Examination Survey. Weight-for-stature and BMI-for-age percentiles were calculated for each child. The 10th and 85th percentiles of weight-for-stature at selected ages were also reexpressed as BMI-for-age percentiles. RESULTS: More than 63% of children had lower weight-for-stature than BMI-for-age percentiles. Children were more likely to be classified as < or = 10th percentile by weight for-stature than by BMI-for-age, but less likely to be classified as > or = 85th percentile. Differences in classification by the 2 measures varied with age and stature and were greater for shorter children. The 10th and 85th percentiles of weight-for-stature corresponded to BMI-for-age percentiles from the 3rd to the 21st percentile and from the 74th to the 92nd percentile, respectively, depending on age and stature. CONCLUSIONS: Weight-for-stature is easier to use than BMI-for age. However, BMI-for-age captures changes in the weight-height relation with age and can be used continuously up to the age of 20 y. BMI-for-age is recommended in most situations. BMI-for-age and weight-for-stature will not give identical results and are not interchangeable. PMID- 11916764 TI - A randomized trial of supplementation with docosahexaenoic acid-rich tuna oil and its effects on the human milk cytokines interleukin 1 beta, interleukin 6, and tumor necrosis factor alpha. AB - BACKGROUND: Increased consumption of n-3 long-chain polyunsaturated fatty acids (PUFAs) has been recommended during pregnancy and lactation. The production of proinflammatory cytokines by human peripheral blood mononuclear cells (PBMCs) can be modified by dietary n-3 PUFAs. OBJECTIVE: We sought to determine whether dietary supplementation of lactating women with docosahexaenoic acid (DHA) can modulate the concentration of cytokines in the aqueous phase of human milk and the production of cytokines by human milk cells (HMCs) and PBMCs. DESIGN: In this double-blind, prospective, randomized trial, mothers of healthy full-term infants were asked to consume daily a nutritional supplement of 2000 mg oil containing either placebo (n = 40), 300 mg DHA + 70 mg eicosapentaenoic acid (EPA; n = 40), or 600 mg DHA + 140 mg EPA (n = 40). The fatty acid composition of plasma, PBMCs, milk, and HMCs from lactating mothers at 4 wk postpartum was measured by gas chromatography. The concentration of interleukin 6 and tumor necrosis factor alpha in milk and the production of interleukin 1beta, tumor necrosis factor alpha, and interleukin 6 by PBMCs and HMCs after stimulation with lipopolysaccharide was measured by enzyme-linked immunosorbent assay. RESULTS: At 4 wk postpartum, 82 mothers were still breast-feeding their infants. DHA increases in maternal plasma, PBMCs, milk, and HMCs were proportional to dietary DHA. There was no relation between tissue DHA status and cytokine concentrations. CONCLUSIONS: Consumption of < or = 600 mg DHA and 140 mg EPA/d for 4 wk increased n-3 PUFA concentrations in relevant tissues but did not cause perturbations in cytokine concentrations in human milk. PMID- 11916766 TI - Both lipid and protein intakes stimulate increased generation of reactive oxygen species by polymorphonuclear leukocytes and mononuclear cells. AB - BACKGROUND: It was recently shown that glucose challenge leads to increased generation of reactive oxygen species (ROS) by polymorphonuclear leukocytes (PMNs) and mononuclear cells (MNCs). OBJECTIVE: To further elucidate the relation between nutrition and ROS generation, we investigated the effect of lipid and protein challenges on ROS generation by leukocytes. DESIGN: After having fasted overnight, one group of healthy subjects consumed a carbohydrate- and protein free cream preparation (1257 kJ) and another group of healthy subjects consumed an equienergetic pure preparation of casein. Sequential blood samples were obtained after the intake of cream and casein. ROS were measured by chemiluminescence after stimulation by N-formyl-methionyl-leucinyl-phenylalanine. Lipid peroxidation was measured as thiobarbituric acid-reactive substances (TBARS) and alpha-tocopherol was measured by HPLC. RESULTS: ROS generation by MNCs and PMNs increased significantly 1, 2, and 3 h after cream intake and 1 h after protein intake. Cholesterol concentrations did not change significantly, whereas triacylglycerol concentrations increased significantly 2 h after cream intake. Total TBARS concentrations increased 1 h after cream intake and remained elevated 3 h after intake, but the increase was not significant when corrected for changes in triacylglycerol. After casein intake, total cholesterol, triacylglycerol, and TBARS concentrations did not change significantly. alpha Tocopherol concentrations did not change significantly after either cream or casein intake. CONCLUSIONS: Both fat and protein intakes stimulate ROS generation. The increase in ROS generation lasted 3 h after cream intake and 1 h after protein intake. Cream intake also caused a significant and prolonged increase in lipid peroxidation. These data are important because increased ROS generation and lipid peroxidation are key events in atherogenesis. PMID- 11916767 TI - Calcium intake influences the association of protein intake with rates of bone loss in elderly men and women. AB - BACKGROUND: There is currently no consensus on the effect of dietary protein intake on the skeleton, but there is some indication that low calcium intakes adversely influence the effect of dietary protein on fracture risk. OBJECTIVE: The objective of the present study was to determine whether supplemental calcium citrate malate and vitamin D influence any associations between protein intake and change in bone mineral density (BMD). DESIGN: Associations between protein intake and change in BMD were examined in 342 healthy men and women (aged > or = 65 y) who had completed a 3-y, randomized, placebo-controlled trial of calcium and vitamin D supplementation. Protein intake was assessed at the midpoint of the study with the use of a food-frequency questionnaire and BMD was assessed every 6 mo by dual-energy X-ray absorptiometry. RESULTS: The mean (+/-SD) protein intake of all subjects was 79.1 +/- 25.6 g/d and the mean total calcium intakes of the supplemented and placebo groups were 1346 +/- 358 and 871 +/- 413 mg/d, respectively. Higher protein intake was significantly associated with a favorable 3-y change in total-body BMD in the supplemented group (in a model containing terms for age, sex, weight, total energy intake, and dietary calcium intake) but not in the placebo group. The pattern of change in femoral neck BMD with increasing protein intake in the supplemented group was similar to that for the total body. CONCLUSION: Increasing protein intake may have a favorable effect on change in BMD in elderly subjects supplemented with calcium citrate malate and vitamin D. PMID- 11916768 TI - Long-chain polyunsaturated fatty acids and diabetes mellitus. PMID- 11916770 TI - Comments on the new dietary reference intake for vitamin E. PMID- 11916771 TI - A trial of labor for remifentanil. PMID- 11916772 TI - Segmental wall motion abnormalities during telerobotic totally endoscopic coronary artery bypass grafting. AB - In addition to single-lung ventilation (SLV), intrathoracic CO2 insufflation is mandatory for adequate exposure during totally endoscopic coronary artery bypass grafting. With transesophageal echocardiography, we investigated biventricular myocardial wall motion in 25 patients with isolated disease of the left anterior descending coronary artery who underwent totally endoscopic coronary artery bypass grafting with the "Da Vinci" robotic surgical system. At distinct time points during the operation, a cine loop of both ventricles was registered from a transgastric mid-short-axis view. Myocardial wall motion analysis was performed according to an established segmentation model of the left ventricle and to an established five-point scale for wall motion (1, normal; 5, dyskinesia). Significant alterations from preoperative baseline wall motion were visible in the septal, inferior, and anterior segments of the left ventricle at some time during the prebypass period, combined with a markedly decreased PaO2 under SLV and increased intrathoracic pressure. The same findings applied to the right ventricle; however, wall motion abnormalities were more pronounced here. After myocardial revascularization, weaning from cardiopulmonary bypass, CO2 deflation, and return to double-lung ventilation, myocardial wall motion recovered to baseline values. Clinically significant hemodynamic instability did not occur. The data suggest that robot-assisted coronary artery bypass grafting leads to significant prebypass alterations of biventricular segmental wall motion. On the basis of our data, it cannot be definitively stated whether the observed results were due to reduced oxygenation during SLV and thus "real" myocardial ischemia, intrathoracic CO2 insufflation with positive pressure leading to mechanical compromise of the heart, absolute or relative hypovolemia, or a combination of these factors. However, in this cohort, which consisted of patients with single vessel disease and good ventricular function, these changes were of limited clinical relevance. IMPLICATIONS: Segmental myocardial wall motion was evaluated with transesophageal echocardiography during robot-assisted totally endoscopic coronary artery bypass grafting. Significant biventricular segmental wall motion abnormalities occurred before cardiopulmonary bypass under single-lung ventilation and carbon dioxide insufflation. The changes in myocardial wall motion were of limited clinical relevance. PMID- 11916773 TI - Does indocyanine green accurately measure plasma volume early after cardiac surgery? AB - Potential overestimation of plasma volume (PV) determination by the conventional indocyanine green (ICG) dilution method (PV-ICG) can occur when generalized capillary protein leakage is present, because ICG binds to proteins. We recently reported that this overestimation can be recognized by simultaneous measurement of the initial distribution volume of glucose (IDVG). We examined whether overestimation of PV-ICG and further ICG-pulse dye densitometry-derived plasma volume (PV-PDD) can occur early after cardiac surgery by using the PV-ICG/IDVG ratio as an indicator. Possible overestimation was defined as a ratio higher than 0.45. Twenty-four consecutive postcardiac surgical patients were enrolled. PV ICG, PV-PDD, and IDVG were calculated simultaneously after admission to the intensive care unit and on the first postoperative day. The mean +/- SD PV ICG/IDVG ratio for 47 recordings was 0.38 +/- 0.05. Four had a PV-ICG/IDVG ratio higher than 0.45, and the highest was 0.48. The mean PV-PDD/IDVG ratio for a total of 47 recordings was 0.39 +/- 0.10. There were extremely high or low ratios observed in PV-PDD determinations, but they were not observed in PV-ICG determinations. Results suggest that most of the PV-ICG measurements are accurate, but inaccuracy of PV-PDD can occur early after cardiac surgery. IMPLICATIONS: Overestimation of indocyanine green-derived plasma volume can occur in the presence of generalized capillary protein leakage. This overestimation was examined early after cardiac surgery by using the simultaneous measurement of the initial distribution volume of glucose. We suggest that overestimation of the traditional dye dilution method is negligible, but apparent over- or underestimation of pulse dye densitometry-derived plasma volume cannot be negligible. PMID- 11916774 TI - The cardiac effects of intracoronary angiotensin II infusion. AB - Angiotensin II (Ang II) is a potent vasoconstrictor, which recently has been shown to also have significant inotropic effects. Previous results regarding the mechanisms of the acute inotropic effects of Ang II are not conclusive. We designed this study to investigate the local cardiac effects of intracoronary Ang II infusion in doses not affecting systemic circulation. Ang II (2.5-40 microg/h) was infused in the left coronary artery of Yorkshire pigs (n = 9) reaching calculated intracoronary Ang II concentrations of 842 +/- 310, 3342 +/- 1238, and 12448 +/- 4393 pg/mL, respectively. Cardiac systolic and diastolic function was evaluated by analysis of the left ventricular pressure-volume relationship. Coronary flow was measured by using a coronary sinus catheter and the retrograde thermodilution technique. No significant changes were seen in the systolic and diastolic function variables of heart rate, end-systolic elastance, preload recruitable stroke work, the time constant for isovolumetric relaxation, or in coronary vascular resistance and flow. The positive inotropic and chronotropic effects of Ang II seen in previous studies seem thus to be mediated via extracardiac actions of Ang II. Coronary vascular tone is not affected by local Ang II infusion in anesthetized pigs. IMPLICATIONS: The positive inotropic and chronotropic effects of angiotension II (Ang II) seen in previous studies seem to be mediated via extracardiac actions of Ang II. Coronary vascular tone is not affected by local Ang II infusion in anesthetized pigs. PMID- 11916775 TI - The feasibility of transesophageal echocardiograph-guided right and left ventricular oximetry in hemodynamically stable patients undergoing coronary artery bypass grafting. AB - There are no techniques available for continuous noninvasive measurement of the oxygen saturation of blood flowing through the heart. We assessed the feasibility and accuracy of transesophageal echocardiograph (TEE)-guided left ventricular (SpO2 LV) and right ventricular (SpO2 RV) oximetry. Twenty hemodynamically stable, well-oxygenated anesthetized patients (ASA physical status III, aged 51 75 yr) undergoing coronary artery bypass grafting were studied. A TEE probe was modified by attaching a single-use pediatric reflectance pulse oximeter just proximal to the ultrasound transducer. The TEE probe was directed toward the LV by using the transgastric mid-short axis view or toward the RV by using the transgastric RV inflow view, in random order. Readings were taken every 30 s for 10 min during a hemodynamically stable period of anesthesia. Simultaneous blood samples were taken from the radial artery and pulmonary artery to determine arterial oxygen saturation (SaO2) and mixed venous oxygen saturation (SvO2), respectively. During SpO2 LV readings, simultaneous finger pulse oximetry (SpO2 finger) was also recorded. SpO2 LV was feasible in 20 of 20 patients, and SpO2 RV was feasible in 19 of 20 patients. The mean +/- SD (range) oxygen saturation for each method was the following: SpO2 LV, 98.7% +/- 0.6% (97%-100%); SaO2, 98.7% +/ 0.6% (96.6%-99.4%); SpO2 finger, 98.1% +/- 1.2% (97%-100%); SpO2 RV, 73.9% +/- 4.7% (64%-85%); and SvO2, 74.5% +/- 4.4% (66.8%-82.6%). SpO2 LV agreed closely with SaO2 (mean difference, 0.072%). SpO2 RV agreed closely with SvO2 (mean difference, 0.65%). SpO2 LV agreed more closely with SaO2 than finger oximetry (mean difference, -0.072 vs -0.692). TEE-guided SpO2 LV and SpO2 RV are feasible in hemodynamically stable anesthetized patients and provide similar readings to arterial and mixed venous blood samples. The technique merits further investigation. IMPLICATIONS: Transesophageal echocardiograph-guided left and right ventricular oximetry is feasible in hemodynamically stable anesthetized patients and provides similar readings to arterial and mixed venous blood samples. PMID- 11916776 TI - The use of bovine hemoglobin glutamer-250 (Hemopure) in surgical patients: results of a multicenter, randomized, single-blinded trial. AB - Hemoglobin-based oxygen carrier-201 (HBOC-201, hemoglobin glutamer-250 [bovine], Hemopure; Biopure Corporation, Cambridge, MA) is polymerized hemoglobin of bovine origin being developed as an oxygen therapeutic. In this study, we evaluated the tolerability of a single intraoperative dose of HBOC-201 in surgical patients. In a single-blinded, multicenter study, 81 patients were randomized to receive either a single infusion of HBOC-201 (55 patients) or an equivalent volume of lactated Ringer's solution (26 patients). Forty-two patients originally assigned to the HBOC-201 group received the entire planned treatment of only one of the following doses: 0.6, 0.9, 1.2, 1.5, 2.0, or 2.5 g/kg of body weight. Thirteen of the 55 patients in the HBOC-201-assigned group did not reach the trigger point for transfusion administration, and they were not included in the analysis. We studied clinical outcomes and compared hematologic findings, blood chemistry values, and blood use in the two treatment groups. There were no patient deaths in this study. No pattern of clinically significant laboratory abnormalities could be attributed to exposure to HBOC-201. In the HBOC-201 group, 2 patients had a transient increased concentration of serum transaminases and 6 had transient skin discoloration. One patient in the HBOC-201 group had mast cell degranulation with hypotension. Postoperatively, methemoglobin plasma concentrations increased in the HBOC-201 group in a dose-dependent manner, reaching maximal values of 3.7% +/- 3.2% (average of all doses given) on postoperative day 3. There was no difference in the mean number of allogeneic blood units transfused in the 2 groups (3.3 +/- 1.8 and 3.7 +/- 4.1 for the lactated Ringer's solution and HBOC-201 groups, respectively) over the course of hospitalization. The intraoperative administration of HBOC-201, up to a maximum of 245 g, was generally well tolerated. There was no relationship between HBOC 201 use and the number of allogeneic blood units transfused over the entire hospitalization course. The administration of HBOC-201 was associated with a delayed (third postoperative day) dose-dependent increase in the plasma methemoglobin concentration. We conclude that the intraoperative use of HBOC-201 was generally well tolerated. IMPLICATIONS: The intraoperative use of hemoglobin glutamer-250 (bovine) (HBOC-201, Hemopure was generally well tolerated. The administration of HBOC-201 was associated with a delayed increase in the plasma methemoglobin concentrations. PMID- 11916777 TI - Preserved gastric tonometric variables in cardiac surgical patients administered intravenous perflubron emulsion. AB - Low gastric intramucosal pH (pHi) and an increased gastric-arterial PCO2 difference (CO2 gap) are markers of tissue hypoperfusion. Perfluorocarbons (PFCs) have a large oxygen-carrying capacity and release oxygen when encountering low tissue oxygen tension. Nine cardiac surgical patients instrumented for gastric tonometry were enrolled as part of a multicenter, randomized, single-blinded study of a PFC emulsion (perflubron emulsion [Oxygent]). Patients were randomized to receive PFC (n = 4) or placebo (n = 5) after intraoperative autologous blood harvesting by acute normovolemic hemodilution. At baseline there were no intergroup differences in tonometric-, hemodynamic-, or oxygen delivery-derived variables, e.g., Control group (pHi, 7.37 +/- 0.06; CO2 gap, 6.4 +/- 1.3 mm Hg) versus PFC group (pHi, 7.38 +/- 0.06; CO2 gap, 6.7 +/- 1.5 mm Hg). After acute normovolemic hemodilution, pHi was significantly lower (P < 0.01) in the Control group (7.22 +/- 0.25) than in the PFC group (7.44 +/- 0.25), and CO2 gap was significantly higher (P < 0.001) in the Control group (23.4 +/- 5.1 mm Hg) than in the PFC group (1.8 +/- 0.8 mm Hg). These differences in tonometric variables persisted during surgery. The PFC group showed a significantly (P < 0.007) shorter time to first bowel movement postoperatively (2.0 +/- 0.8 vs 5.4 +/- 1.6 days). Time to consumption of solid food was also shorter in the PFC group and almost achieved statistical significance (P = 0.056). IMPLICATIONS: This study suggests that the administration of perflubron emulsion prevents gastrointestinal tract ischemia in cardiac surgical patients and may preserve postoperative gastrointestinal tract function. PMID- 11916778 TI - The reduction of preoperative autologous blood donation for primary total hip or knee arthroplasty: the effect on subsequent transfusion rates. AB - We conducted this quality assurance observational study to examine the effects of a change in policy regarding preoperative autologous blood donation (PABD) and indications for perioperative blood transfusion in patients undergoing primary total hip or knee arthroplasty. Two successive time periods, each including 182 successive patients treated by the same medical team and with standardized anesthesia, were compared. The first study had the following standard transfusion policy: 3 U of PABD collected (n = 119) and liberal autologous transfusion (AT). The second study introduced a specific indication for PABD, on the basis of estimated red blood cell reserve and a life expectancy of more than 10 years; 2 U of PABD was collected (n = 81), and criteria were identical for AT and allogeneic transfusion. We mainly compared the incidence of AT; allogeneic and overall transfusions; the inclusion, admission, and discharge hematocrit values; and the wastage of PABD units. This novel policy increased the number of untransfused patients by a factor of 10 (5.5% vs 56.6%) (P < 0.0001), decreased the number of PABD patients by 30% with a 2.4-fold reduction in AT (30% vs 80%) (P < 0.0001), and did not change allogeneic requirements (13% vs 15%). Although fewer autologous units were collected (172 vs 426), the wastage was higher in Study 2 (46% vs 12%) (P < 0.0001). We conclude that incorporation of patients' individual factors improves the efficiency of transfusion for total hip and total knee arthroplasty surgeries. IMPLICATIONS: We compared two transfusion policies for primary total hip or knee arthroplasties: first, a standard preoperative autologous donation with a liberal autologous transfusion policy; and second, a more restrictive indication for autologous donation that was based on patients' individual factors, with identical criteria for autologous and allogeneic transfusion. We found that this change of policy reduced autologous donation and transfusion with no increase in allogeneic transfusion. PMID- 11916779 TI - The effect of methylene blue on the hemodynamic changes during ischemia reperfusion injury in orthotopic liver transplantation. AB - After graft reperfusion in orthotopic liver transplantation (OLT), ischemia reperfusion syndrome (IRS) is characterized by persistent hypotension with a low systemic vascular resistance. Methylene blue (MB) has been used as a vasopressor in sepsis and acute liver failure. We investigated the effect of MB on IRS during OLT. Thirty-six patients undergoing elective OLT were randomized to receive either a bolus of MB 1.5 mg/kg before graft reperfusion, or normal saline (placebo). We recorded hemodynamic variables, postoperative liver function tests, and time to hospital discharge. Blood samples were analyzed for arterial lactate concentration, cyclic 3',5'-monophosphate, and plasma nitrite/nitrate concentrations. The MB group had higher mean arterial pressure (P = 0.035), higher cardiac index (P = 0.04), and less epinephrine requirement (P = 0.02). There was no difference in systemic vascular resistance or central venous pressure. Serum lactate levels were lower at 1 h after reperfusion in MB patients, suggesting better tissue perfusion (P = 0.03). In the presence of MB, there was a reduction in cyclic 3',5'-monophosphate (P < 0.001), but not plasma nitrites. Postoperative liver function tests and time to hospital discharge were the same in both groups. MB attenuated the hemodynamic changes of IRS in OLT acting via guanylate cyclase inhibition. IMPLICATIONS: Methylene blue attenuates the hemodynamic changes of the ischemia reperfusion syndrome in liver transplantation, and this effect involves guanylate cyclase inhibition. PMID- 11916780 TI - The effects of intravenous almitrine on oxygenation and hemodynamics during one lung ventilation. AB - One-lung ventilation (OLV) induces an increase in pulmonary shunt sometimes associated with a decrease in PaO2 despite ventilation with 100% oxygen. PaO2 improvement has been reported in one-lung ventilated animals receiving IV almitrine, a pulmonary vasoconstrictor. We evaluated the ability of almitrine to prevent a decrease in PaO2 during OLV. Patients without pulmonary hypertension undergoing OLV for lung surgery were randomly assigned to receive either placebo (Group P, n = 8) or almitrine infusion at a rate of 8 microg x kg(-1) x min(-1) (Group A, n = 8) from the start of OLV. Gasometric and hemodynamic values were recorded with the patient in the lateral decubitus position during two-lung ventilation and at 10-min intervals during OLV over a 30-min period (OLV-10, OLV 20, OLV-30). Compared with the values found during two-lung ventilation (434 +/- 22 mm Hg in Group P and 426 +/- 23 mm Hg in Group A), PaO2 decreased at OLV-10 (305 +/- 46 mm Hg), OLV-20 (203 +/- 20 mm Hg), and OLV-30 (178 +/- 18 mm Hg) in Group P (P < 0.05) and at OLV-20 (354 +/- 25 mm Hg) and OLV-30 (325 +/- 17 mm Hg) in Group A (P < 0.05). PaO2 values differed between the groups at OLV-20 and OLV 30 (P < 0.05). Pulmonary artery pressure and cardiac output did not change. In conclusion, 8 microg x kg(-1) x min(-1) IV almitrine prevents and limits the OLV induced decrease in PaO2 without causing any hemodynamic modification. IMPLICATIONS: Eight microg x kg(-1) x min(-1) IV almitrine limits one-lung ventilation-induced decrease in PaO2 without causing any hemodynamic modification in patients without pulmonary hypertension. PMID- 11916781 TI - Terlipressin-ephedrine versus ephedrine to treat hypotension at the induction of anesthesia in patients chronically treated with angiotensin converting-enzyme inhibitors: a prospective, randomized, double-blinded, crossover study. AB - In patients chronically treated with angiotensin converting-enzyme inhibitors (ACEI), typically selected doses of ephedrine do not always restore arterial blood pressure when anesthesia-induced hypotension occurs. We postulated that the administration of terlipressin, an agonist of the vasopressin system, with ephedrine more effectively restores pressure in this setting than the administration of ephedrine alone. This prospective, randomized, cross-over, double-blinded study compared terlipressin combined with ephedrine (n = 19) with ephedrine alone (n = 21) in treating hypotension at the induction of anesthesia in 40 ACEI-treated patients undergoing hypotension (mean arterial blood pressure [MAP] <65 mm Hg or <30% of baseline value) after standardized anesthetic protocol (target-controlled IV anesthesia with propofol). Data are mean +/- SD. Patient characteristics, MAP, and heart rate before and after the induction of anesthesia during hypotensive episodes were not significantly different between the two groups. After the first bolus, MAP was significantly greater in the Terlipressin Ephedrine group (72 +/- 12 mm Hg versus 65 +/- 8 mm Hg, P < 0.05). The occurrence of a second hypotensive episode (5% versus 71%, P < 0.001), the duration (2 +/- 1 min versus 3 +/- 1 min, P < 0.01) of hypotensive episodes, and the median dose of ephedrine (3 versus 6 mg, P < 0.05) were significantly less in the Terlipressin Ephedrine group. In conclusion, terlipressin combined with ephedrine is more effective than ephedrine alone for treating anesthesia-induced hypotension in ACEI-treated patients. We conclude that this patient population with a partially blocked endogenous response to hypotension may be good candidates for successful use of a vasopressin analog to counteract intraoperative refractory hypotension. IMPLICATIONS: Vascular surgical patients chronically treated with drugs that inhibit the functioning of the renin-angiotensin system may experience hypotension unresponsive to conventional therapy. This double-blinded, cross-over study demonstrated that in these patients the use of a vasopressin analog, terlipressin given with ephedrine, was effective in reversing intraoperative systemic hypotension refractory to ephedrine. PMID- 11916782 TI - The role of mitochondrial and sarcolemmal K(ATP) channels in canine ethanol induced preconditioning in vivo. AB - Chronic consumption of small doses of ethanol protects myocardium from ischemic injury. We tested the hypothesis that mitochondrial and sarcolemmal adenosine triphosphate-dependent potassium (K(ATP)) channels mediate these beneficial effects. Dogs (n = 76) were fed with ethanol (1.5 g/kg) or water mixed with dry food bid for 6 or 12 wk, fasted overnight before experimentation, and instrumented for measurement of hemodynamics. Dogs received intracoronary saline (vehicle), 5-hydroxydecanoate (a mitochondrial K(ATP) channel antagonist; 6.75 mg/kg over 45 min), or HMR-1098 (a sarcolemmal K(ATP) channel antagonist; 45 microg/kg over 45 min) and were subjected to a 60 min coronary artery occlusion followed by 3 h of reperfusion. A final group of dogs was pretreated with ethanol and chow for 6 wk before occlusion and reperfusion. Myocardial infarct size and transmural coronary collateral blood flow were measured with triphenyltetrazolium chloride staining and radioactive microspheres, respectively. The area at risk of infarction was similar between groups. A 12-wk pretreatment with ethanol significantly reduced infarct size to 13% +/- 2% (mean +/- SEM; n = 8) of the area at risk compared with control experiments (25% +/- 2%; n = 8), but a 6-wk pretreatment did not (21% +/- 2%; n = 8). 5-hydroxydecanoate and HMR-1098 abolished the protective effects of 12-wk ethanol pretreatment (24% +/- 2% and 29% +/- 3%, respectively; n = 8 for each group) but had no effect in dogs that did not receive ethanol (22% +/- 2% and 23% +/- 4%, respectively; n = 8 for each group). No differences in hemodynamics or transmural coronary collateral blood flow were observed between the groups. The results indicate that mitochondrial and sarcolemmal K(ATP) channels mediate ethanol-induced preconditioning in dogs independent of alterations in systemic hemodynamics or coronary collateral blood flow. IMPLICATIONS: Mitochondrial and sarcolemmal K(ATP) channels mediate ethanol induced preconditioning independent of alterations in systemic hemodynamics or coronary collateral perfusion in vivo. PMID- 11916783 TI - Isoflurane inhibits neutrophil-endothelium interactions in the coronary circulation: lack of a role for adenosine triphosphate-sensitive potassium channels. AB - Isoflurane protects myocardium during ischemia-reperfusion via a mechanism involving the adenosine triphosphate-sensitive potassium channels. We tested the hypothesis that an inhibition of the neutrophil-endothelium interactions by isoflurane contributes to this effect. Polymorphonuclear neutrophils and coronary artery segments were obtained from 35 healthy dogs. Superoxide production by neutrophils, stimulated with platelet-activating factor (PAF; 1.0 microM), was measured spectrophotometrically. Adherence of PAF-activated neutrophils to the endothelium of coronary segments was assessed by direct counting of neutrophils labeled with fluorescent dye. Coronary artery rings were exposed to PAF-activated neutrophils, and, after washing and preconstriction with U46619, they were evaluated for vasorelaxation responses to acetylcholine (endothelium dependent) and sodium nitroprusside (endothelium independent). Measurements were performed in the absence and presence of isoflurane (1 and 2 minimum alveolar anesthetic concentration) both with and without glibenclamide (10 microM). Isoflurane inhibited superoxide production and adherence by neutrophils and abolished neutrophil-induced reductions in coronary vascular relaxation responses to acetylcholine. Glibenclamide did not alter the effects of isoflurane on neutrophils or coronary artery endothelium. In conclusion, isoflurane had an inhibitory action on neutrophil-endothelium interactions and neutrophil-mediated coronary endothelial dysfunction--effects that may be involved in its cardioprotective action in vivo. These inhibitory actions of isoflurane were not mediated by adenosine triphosphate-sensitive potassium channels. IMPLICATIONS: Isoflurane inhibited neutrophil-endothelium interactions and the inflammatory response in vitro via a pathway independent of the adenosine triphosphate sensitive potassium channels. This action could be involved in the cardioprotection by isoflurane observed in vivo. PMID- 11916784 TI - Prosthetic valve malfunction masked by intraoperative pressure measurements. AB - IMPLICATIONS: We describe a case in which intraoperative echocardiography recorded an abnormally high pressure gradient across a newly implanted mechanical heart valve. However, inserting pressure-transducing needles on each side of this prosthesis did not confirm the echocardiographic findings. The prosthesis was later confirmed to be malfunctioning and was replaced. PMID- 11916786 TI - The reduction in minimum alveolar concentration for tracheal extubation after clonidine premedication in children. AB - The effects of clonidine on minimum alveolar concentration for tracheal extubation (MAC-ex) have not been elucidated. Clonidine may lead to prolonged emergence from anesthesia. We investigated the effects of oral clonidine premedication on MAC-ex and examined the emergence properties of sevoflurane in children. Sixty ASA physical status I pediatric patients, aged from 2 to 9 yr, were randomly divided into one of three groups and received placebo, clonidine 2 microg/kg, or clonidine 4 microg/kg (n = 20 each) orally, 100 min before the induction of anesthesia. The induction of anesthesia, tracheal intubation, and maintenance of anesthesia were performed with sevoflurane in air and oxygen. MAC ex was defined according to the modification of Dixon's up-and-down method, with 0.25% as a step size. In addition, in the Control and 4 microg/kg groups, the time from tracheal extubation to spontaneous eye opening (eye-opening time) and the time from tracheal extubation to leaving the operating room (awakening time) were recorded. MAC-ex for sevoflurane (mean +/- SD) was 1.63% +/- 0.13%, 1.04% +/ 0.26%, and 0.66% +/- 0.09% respectively in the Control group, 2 microg/kg group, and 4 microg/kg group. Significant differences were observed among the three groups. The eye-opening times were 5.7 +/- 3.5 min in the Control group and 5.1 +/- 1.0 min in the 4 microg/kg group. The awakening times were 9.7 +/- 3.7 min in the Control group and 9.2 +/- 3.8 min in the 4 microg/kg group. No significant differences were observed among the groups. IMPLICATIONS: Oral clonidine premedication decreased MAC for tracheal extubation for sevoflurane dose dependency and did not prolong emergence from anesthesia. PMID- 11916785 TI - Postoperative pain relief in children undergoing tympanomastoid surgery: is a regional block better than opioids? AB - Peripheral nerve blocks of the surgical site can reduce the need for perioperative opioids thereby decreasing their unwanted adverse effects, such as postoperative nausea and vomiting. In this prospective, randomized, double blinded study, we examined the efficacy of a great auricular nerve (GAN) block compared with IV morphine sulfate in children undergoing tympanomastoid surgery. After the induction of general anesthesia, children were randomized to receive either a GAN block with 2 mL of 0.25% bupivacaine with epinephrine (1:200,000) and a sham IV injection of 2 mL of saline solution or a sham GAN block with 2 mL of saline solution with an IV injection of 0.1 mg/kg morphine sulfate diluted to 2 mL. Patients' objective pain scores were assessed by a blinded observer and the incidence of vomiting was recorded. The GAN-Block patients as a group required more pain rescue in the postanesthesia care unit; this difference was not statistically different from the IV-morphine group (P = 0.084). Nine GAN-Block patients never received opioid or other analgesics at any time in the first 24 h after surgery. The group that received the GAN block also had a less frequent incidence of vomiting requiring intervention (7 versus 19) during their entire hospitalization or at home (P = 0.027). The GAN-Block group also had more patients who never experienced vomiting (13 of 20 versus 5 of 20, P = 0.026). In this cohort, a peripheral nerve block decreased the overall incidence of postoperative vomiting thereby reducing associated costs. IMPLICATIONS: We prospectively compared the use of a great auricular nerve block versus IV morphine sulfate in a randomized double-blinded study in children undergoing tympanomastoid surgery. Analgesia was comparable between groups but nearly half the Block group never required additional analgesics and the number of vomiting events was nearly 66% less. PMID- 11916787 TI - A study of lidocaine iontophoresis for pediatric venipuncture. AB - In this randomized, double-blinded, placebo-controlled study, we evaluated the safety and efficacy of lidocaine iontophoresis for the prevention of pain associated with venipuncture in 59 children aged 6-17 yr. Children received either lidocaine HCl 2% with epinephrine 1:100,000 (Active) or the same formulation without lidocaine (Placebo) via a 20 mA/min iontophoretic treatment. Pain during venipuncture was assessed by the subject, parent, and nurse using a 100-mm visual analog scale. Median (interquartile range) visual analog scale scores were significantly lower in the Active versus Placebo groups: subject, 7.0 (2.0-20.8) versus 31.0 (12.0-48.0), P < 0.001; nurse, 5.0 (2.2-10.8) versus 24.0 (9.0-47.0), P < 0.001; and parent, 3.0 (0.8-7.2) versus 20.0 (4.5-43.0), P < 0.002, respectively. Similarly, higher median satisfaction scores were given to the Active versus Placebo group by the three evaluators. Of the 59 subjects completing the study, 10 subjects experienced a total of 12 adverse events that were all graded as mild. In conclusion, lidocaine iontophoresis is safe in children, reduces discomfort associated with venipuncture, and increases satisfaction when compared with the placebo. IMPLICATIONS: In this randomized, double-blinded, placebo-controlled study, we found that dermal anesthesia with lidocaine HCl 2% combined with epinephrine 1:100,000 administered via iontophoresis in children is achieved in 8.8 +/- 2.1 min, reduces discomfort associated with venipuncture, is safe, and increases satisfaction when compared with the placebo. PMID- 11916788 TI - Korean hand acupressure reduces postoperative nausea and vomiting after gynecological laparoscopic surgery. AB - To investigate the effectiveness of prophylactic Korean hand acupressure in the prevention of postoperative vomiting in women scheduled for minor gynecological laparoscopic surgery, we conducted a double-blinded, randomized, placebo controlled study. In one group (n = 40), acupressure was performed 30 min before the induction of anesthesia by using special acupressure seeds, which were fixed onto the Korean hand acupuncture point K-K9 and remained there for at least 24 h. The second group (n = 40) functioned as the Placebo group. The treatment groups did not differ with regard to demographics, surgical procedure, or anesthetic administered. In the Acupressure group, the incidence of nausea and vomiting was significantly less (40% and 22.5%) than in the Placebo group (70% and 50%). We conclude that Korean hand acupressure of the acupuncture point K-K9 is an effective method for reducing postoperative nausea and vomiting in women after minor gynecological laparoscopic surgery. IMPLICATIONS: This randomized study was performed to investigate the antiemetic effect of the Korean hand acupuncture point K-K9. Acupressure of K-K9 reduces the incidence of postoperative nausea and vomiting in female patients after minor gynecological laparoscopic surgery. PMID- 11916789 TI - A comparison of neuromuscular effects, tracheal intubating conditions, and reversibility of rapacuronium versus mivacurium in female patients. AB - IMPLICATIONS: Rapacuronium is a new, rapid-onset, short-duration, nondepolarizing neuromuscular blocking drug. We evaluated the intubating conditions at maximum block after the administration of rapacuronium or mivacurium in female patients undergoing laparoscopy. We also evaluated the neostigmine-induced reversibility of neuromuscular block after this single dose of rapacuronium or mivacurium. PMID- 11916791 TI - The interaction between propofol and clonidine for loss of consciousness. AB - Clonidine premedication reduces the intraoperative requirement for opioids and volatile anesthetics. Clonidine also reduces the induction dose of IV anesthetics. There is no information, however, regarding the effect of oral clonidine premedication on the propofol blood concentrations required for loss of consciousness, and the interaction between propofol and clonidine. We randomly administered target effect-site concentrations of propofol ranging from 0.5 to 5. 0 microg/mL by using computer-assisted target-controlled infusion to 3 groups of healthy male patients: Control (n = 35), 2.5 microg/kg Clonidine (n = 36), and 5.0 microg/kg Clonidine (n = 36) groups. Nothing was administered to the Control group. Clonidine (2.5 or 5.0 microg/kg) was administered orally 90 min before the induction of anesthesia in the Clonidine groups. After equilibration between the blood and effect-site for 15 min, a verbal command to open their eyes was given two times to the patients. Arterial blood samples for analysis of the serum propofol and clonidine concentrations were taken immediately before verbal commands were given. Measured serum propofol concentrations in equilibrium with the effect-site at which 50% of the patients did not respond to verbal commands (EC50 for loss of consciousness) were determined by logistic regression. The EC50 +/- SE values in the Control, 2.5 microg/kg Clonidine, and 5.0 microg/kg Clonidine groups were 2.67 +/- 0.18, 1.31 +/- 0.12, and 0.91 +/- 0.13 microg/mL, respectively. The EC50 in the 2.5 and 5.0 microg/kg clonidine groups was significantly smaller than that in the Control group (P < 0.001). The use of a response surface modeling analysis indicated that there was an additive interaction between measured arterial propofol and clonidine concentrations in relation to loss of consciousness. These results indicate that propofol and clonidine act additively for loss of consciousness. IMPLICATIONS: Oral clonidine 2.5 and 5.0 microg/kg premedication decreases the propofol concentration required for loss of consciousness. PMID- 11916790 TI - Neuromuscular pharmacology of TAAC3, a new nondepolarizing muscle relaxant with rapid onset and ultrashort duration of action. AB - We selected bis [N-(3,4-diacetoxybenzyl) tropanium-3alpha-yl] glutarate dibromide (TAAC3) from many new tropinyl diester derivatives to evaluate its neuromuscular blocking (NMB) and autonomic side effects on anesthetized rats, rabbits, guinea pigs, cats, pigs, dogs, and monkeys. NMB potency, onset, recovery index, and duration of action were determined. Comparisons of these pharmacologic variables were made between TAAC3 and rocuronium. In the cat, the degrees of train-of-four and tetanic fade, posttetanic potentiation, and pharmacologic antagonism were evaluated. For determination of the NMB maintenance dose, TAAC3 was also given to rabbits and pigs in the initial dose/maintenance infusion mode. Cardiac vagal block was evaluated in the rat, pig, cat, and guinea pig on the basis of the inhibition of the bradycardia to stimulation of the vagus nerve. Sympathetic ganglion block was studied on the superior cervical ganglion-nictitating membrane preparation of the cat. TAAC3 produced nondepolarizing NMB. Its NMB 90% effective doses ranged from 90 to 425 microg/kg, depending on the species. TAAC3 had a faster onset (0.8-1.0 min), shorter recovery index (0.6-1.1 min), and shorter duration of action (1.8-3.5 min) than rocuronium. It produced a slight cumulative effect on infusion, but not on repeated single-dose administration. Cardiac vagal block was present at doses exceeding the NMB 90% effective dose. In the cat and pig at equipotent NMB doses, the degree of cardiac vagal block was similar to that of rocuronium. There was no demonstrable sympathetic ganglion block in the cat. In view of its favorable NMB characteristics, TAAC3 is now undergoing detailed preclinical studies. IMPLICATIONS: We developed a new nondepolarizing muscle relaxant, TAAC3, and investigated it in several animal models. TAAC3 has shown a very rapid onset and an ultrashort duration of neuromuscular blocking action. A minor degree of cardiac vagal block was observed. TAAC3 is promising for further studies. PMID- 11916792 TI - The effects of propofol on neural and endothelial control of in situ rat mesenteric vascular smooth muscle transmembrane potentials. AB - We indirectly assessed the in vivo effect of propofol on sympathetic neural and endothelial control of vascular smooth muscle (VSM) tone in Sprague-Dawley rats by measurement of in situ responses of VSM transmembrane potential (E(m)) in intact, small mesenteric arteries and veins superfused with physiologic salt solution. Measurements were made before, during, and after propofol infusion (10 and 30 mg x kg(-1) x h(-1)) in sympathetically innervated and locally denervated vessels. Propofol's effect on E(m) response to superfusion with acetylcholine (ACh), in physiologic salt solution also containing NG-nitro-L-arginine-methyl ester and indomethacin, was determined in innervated vessels. At 30 mg x kg(-1) x h(-1), propofol caused greater arterial VSM hyperpolarization in innervated compared with denervated vessels (4.8 +/- 2.0 mV versus 2.8 +/- 1.5 mV, respectively). ACh hyperpolarized arterial, but not venous, VSM (e.g., 11.7 +/- 2.4 mV at 10(-4) M). ACh-induced hyperpolarization was eliminated by 30 mg x kg( 1) x h(-1) propofol. Assuming a close inverse correlation between magnitude of VSM E(m) and contractile force, these results suggest that propofol attenuates both sympathetic neural and nonneural regulation of VSM tone. They also suggest that propofol and ACh may act competitively in the second messenger cascade regulating VSM K+ channel activity in mesenteric resistance arteries. IMPLICATIONS: Vascular smooth muscle (VSM) contractile force responses to the IV anesthetic, propofol, were indirectly assessed by VSM membrane potential changes to clarify the mechanisms underlying attenuation of peripheral vascular control of arterial blood pressure. Results indicate that propofol-induced VSM membrane hyperpolarization and coupled reduction of VSM contractile force underlie such attenuation. PMID- 11916793 TI - The stability of a ketamine-morphine solution. AB - Recent advances in acute pain mechanisms and management have implicated the N methyl D-aspartate receptor-ion channel complex in the development of postoperative hyperalgesia and acute opioid tolerance. N-methyl D-aspartate receptor antagonists such as ketamine have been used increasingly in clinical studies in an effort to minimize acute postoperative pain and reduce opioid requirements. A mixture of ketamine and an opioid administered in the same solution and syringe would be a practical and useful technique for postoperative epidural analgesia, continuous IV infusion, or patient-controlled IV analgesia. We investigated the stability of a morphine sulfate and racemic ketamine solution in saline at pH 5.5-7.5 over a period of 4 days. Our study demonstrates that the ketamine-morphine mixture at a clinically relevant concentration seems to be stable at room temperature, at a wide range of pH values, for at least 4 days. IMPLICATIONS: Small-dose ketamine is used with increasing frequency in the acute postoperative setting as an adjunct to traditional opioid analgesics. We show that a racemic ketamine and morphine solution at a clinically relevant concentration seems to be stable at room temperature at a wide range of pH values for at least 4 days. PMID- 11916794 TI - Tramadol inhibits norepinephrine transporter function at desipramine-binding sites in cultured bovine adrenal medullary cells. AB - Tramadol is a widely used analgesic, but its mode of action is not well understood. To study the effects of tramadol on norepinephrine transporter (NET) function, we assayed the effect of tramadol on [3H]-norepinephrine ([3H]-NE) uptake and [3H]-desipramine binding to plasma membranes isolated from bovine adrenal medulla. We then characterized [14C]-tramadol binding in cultured bovine adrenal medullary cells. Tramadol inhibited the desipramine-sensitive uptake of [3H]-NE by the cells in a concentration-dependent manner (50% inhibitory concentration = 21.5 +/- 6.0 microM). Saturation analysis revealed that tramadol increased the apparent Michaelis constant of [3H]-NE uptake without changing the maximal velocity, indicating that inhibition occurred via competition for the NET (inhibition constant, K(i) = 13.7 microM). Tramadol inhibited the specific binding of [3H]-desipramine to plasma membranes. Scatchard analysis of [3H] desipramine binding revealed that tramadol increased the apparent dissociation constant (K(d)) for binding without altering maximal binding, indicating competitive inhibition (K(i) = 11.2 microM). The binding of [14C]-tramadol to the cells was specific and saturable, with a K(d) of 18.1 +/- 2.4 microM. These findings indicate that tramadol competitively inhibits NET function at desipramine-binding sites. IMPLICATIONS: Tramadol competitively inhibits norepinephrine transporter function at desipramine-binding sites in the adrenal medullary cells and probably the noradrenergic neurons of the descending inhibitory system. PMID- 11916795 TI - Motor-evoked potential facilitation during progressive cortical suppression by propofol. AB - We characterized the effects of various stimulation patterns on motor-evoked potentials (MEPs) elicited by repetitive transcranial magnetoelectric stimulation at different levels of cortical suppression by propofol. In 20 patients undergoing lumbar disk surgery, propofol target plasma concentrations (PTPCs) were increased incrementally by target plasma-level controlled infusion during the induction of anesthesia. MEPs were recorded from the muscles of the upper extremities after single, double, and quadruple magnetoelectric stimulation at 500, 200, and 100 Hz. The mean PTPC during loss of responsiveness to verbal instructions (CP50) was 3 microg/mL (CP(95), 5 microg/mL). At PTPCs <3 microg/mL, maximal MEP amplitudes were elicited by quadruple stimulation at 100 Hz. At PTPCs > or =3 microg/mL, four pulses at 200 Hz yielded peak MEP amplitudes. Therefore, quadruple magnetoelectric stimulation at 100 Hz yields peak myogenic responses in awake patients. With progressive cortical suppression resulting from PTPCs beyond 3 microg/mL, the most effective stimulation frequency shifts to 200 Hz. This may be explained by a differential dose-dependent action of propofol on GABAergic cortical interneurons, corresponding to the clinically observed state of vigilance. Recording of spinal cord evoked potentials will aid in further elucidation of the modulatory effects of general anesthesia on intracortical facilitation. IMPLICATIONS: We investigated the effect of different transcranial magnetoelectric stimulation paradigms on motor-evoked potentials during different levels of cortical suppression by propofol. The most effective stimulation frequency seems to change from 100 to 200 Hz during progressive propofol dose escalation, possibly because of specific interaction with cortical interneurons. PMID- 11916796 TI - Remifentanil in obstetric analgesia: a dose-finding study. AB - IV patient-controlled analgesia (PCA) with remifentanil is a new approach in systemic opioid analgesia during labor. We determined the minimum effective dose of IV remifentanil by increasing the PCA bolus from 0.2 microg/kg with 0.2 microg/kg increments during a 60-min study period until the analgesia was considered adequate by the parturient. Twenty healthy parturients with singleton pregnancies participated in the study during the first stage of labor. Remifentanil hydrochloride was given IV via PCA over 1 min with a lockout time of 1 min. The parturient started the PCA bolus at the first subjective sign of uterine contraction. All 17 patients who completed the study reached adequate pain relief. The median effective PCA bolus was 0.4 microg/kg and consumption was 0.066 microg x kg(-1) x min(-1), with wide individual variation (0.2-0.8 microg/kg and 0.027-0.207 microg x kg(-1) x min(-1), respectively). The pain scores were reduced by a median of 4.2 (25th-75th percentiles, 3.1-5.2; P < 0.001) on an 11-point numeric scale. Although there was a wide individual variation in the dose required, remifentanil seems effective for labor analgesia. However, maternal oxygen desaturation, sedation, and reduced fetal heart rate beat-to-beat variability were observed frequently. There was wide individual variation in the dose required for effective labor analgesia. Potentially serious side effects, which were observed frequently during remifentanil analgesia, may limit remifentanil's use in obstetrics. IMPLICATIONS: We determined the minimum effective dose of patient-controlled IV remifentanil for labor analgesia. There was wide individual variation in the dose required for effective labor analgesia. Potentially serious side effects, which were observed frequently during remifentanil analgesia, may limit its use in obstetrics. PMID- 11916797 TI - Prolonged intravenous remifentanil infusion for labor analgesia. AB - IMPLICATIONS: A 34-h remifentanil infusion was administered for labor analgesia in a patient with thrombocytopenia and renal insufficiency. Compared with other opioids, remifentanil may produce fewer cumulative effects during prolonged infusion because of its unique metabolism. PMID- 11916798 TI - A quantitative, systematic review of randomized controlled trials of ephedrine versus phenylephrine for the management of hypotension during spinal anesthesia for cesarean delivery. AB - This quantitative systematic review compared the efficacy and safety of ephedrine with phenylephrine for the prevention and treatment of hypotension during spinal anesthesia for cesarean delivery. Seven randomized controlled trials (n = 292) were identified after a systematic search of electronic databases (MEDLINE, EMBASE, The Cochrane Controlled Trials Registry), published articles, and contact with authors. Outcomes assessed were maternal hypotension, hypertension and bradycardia, and neonatal umbilical cord blood pH values and Apgar scores. For the management (prevention and treatment) of maternal hypotension, there was no difference between phenylephrine and ephedrine (relative risk [RR] of 1.00; 95% confidence interval [CI], 0.96-1.06). Maternal bradycardia was more likely to occur with phenylephrine than with ephedrine (RR of 4.79; 95% CI, 1.47-15.60). Women given phenylephrine had neonates with higher umbilical arterial pH values than those given ephedrine (weighted mean difference of 0.03; 95% CI, 0.02-0.04). There was no difference between the two vasopressors in the incidence of true fetal acidosis (umbilical arterial pH value of <7.2; RR of 0.78; 95% CI, 0.16 3.92) or Apgar score of <7 at 1 and 5 min. This systematic review does not support the traditional idea that ephedrine is the preferred choice for the management of maternal hypotension during spinal anesthesia for elective cesarean delivery in healthy, nonlaboring women. IMPLICATIONS: Phenylephrine and ephedrine to manage hypotension during spinal anesthesia for elective cesarean delivery were compared in this systematic review. Women given ephedrine had neonates with lower umbilical cord blood pH values compared with those given phenylephrine. However, no differences in the incidence of fetal acidosis (pH value of <7.2) or neonatal Apgar scores were found. PMID- 11916800 TI - How to schedule elective surgical cases into specific operating rooms to maximize the efficiency of use of operating room time. AB - We considered elective case scheduling at hospitals and surgical centers at which surgeons and patients choose the day of surgery, cases are not turned away, and anesthesia and nursing staffing are adjusted to maximize the efficiency of use of operating room (OR) time. We investigated scheduling a new case into an OR by using two patient-scheduling rules: Earliest Start Time or Latest Start Time. By using several scenarios, we showed that the use of Earliest Start Time is rational economically at such facilities. Specifically, it maximizes OR efficiency when a service has nearly filled its regularly scheduled hours of OR time. However, Latest Start Time will perform better at balancing workload among services' OR time. We then used historical case duration data from two facilities in computer simulations to investigate the effect of errors in predicting case durations on the performance of these two heuristics. The achievable incremental reduction in overtime by having perfect information on case duration versus using historical case durations was only a few minutes per OR. The differences between Earliest Start Time and Latest Start Time were also only a few minutes per OR. We conclude that for facilities at which the goals are, in order of importance, safety, patient and surgeon access to OR time, and then efficiency, few restrictions need to be placed on patient scheduling to achieve an efficient use of OR time. IMPLICATIONS: We showed how elective cases should be scheduled to maximize the efficiency of use of operating room time. The analysis applies to surgical suites at which surgeons and patients have access to operating room time every workday. PMID- 11916799 TI - A comparison of epidural infusions in the combined spinal/epidural technique for labor analgesia. AB - We compared the clinical effects of three epidural infusions initiated after subarachnoid medication was administered as part of the combined spinal/epidural technique for labor analgesia. Fifteen minutes after administering subarachnoid fentanyl 25 microg and 1 mL of bupivacaine 0.25%, and 5 min after an epidural test dose of 3 mL of bupivacaine 0.25%, women were randomized to receive an epidural infusion of saline, bupivacaine 0.125%, bupivacaine 0.0625%, or bupivacaine 0.04% with epinephrine 1:600,000. All epidural infusions were started at 10 mL/h, and all except the Saline Group also received fentanyl 2 microg/mL. The end point of the study was delivery or request for additional medication for analgesia. We found that time until request for additional analgesia was longest in women who received bupivacaine 0.125% (median duration, 300 min) versus saline (median duration, 118 min) (P = 0.0001) and was intermediate for bupivacaine 0.0625% and bupivacaine 0.04% (median duration, 162 and 180 min, respectively) (P = 0.0001 versus saline). Women who received bupivacaine 0.125% had the most motor block. We conclude that all the bupivacaine-based infusions we tested maintained the analgesia from subarachnoid medication longer than saline, with the longest duration, but the most motor block, from bupivacaine 0.125%. IMPLICATIONS: In this prospective, randomized, and double-blinded study we found that initiating an epidural infusion of bupivacaine 0.125% with fentanyl 2 microg/mL at 10 mL/h 15 min after subarachnoid fentanyl 25 microg with 1 mL of bupivacaine 0.25%, followed by an epidural test dose of 3 mL of bupivacaine 0.25%, maintained the analgesia for longer but with more motor block than with either bupivacaine 0.04% or bupivacaine 0.0625%. PMID- 11916801 TI - Halothane potentiation of hydrogen peroxide-induced inhibition of surfactant synthesis: the role of type II cell energy status. AB - Small concentrations of inhaled anesthetics can affect Type II cell surfactant production and exacerbate oxidant-mediated lung injury. We hypothesized that inhaled anesthetics augment oxidant-induced Type II pneumocyte dysfunction related to their different effects on cellular adenosine triphosphate (ATP) status. Freshly isolated Type II cells were exposed to different concentrations of hydrogen peroxide (H2O2) in the presence or absence of an in vitro halothane exposure. Cells exposed to 100 microM H2O2 alone demonstrated a 23% decrease in ATP levels and a 32% decrease in phosphatidylcholine (PC) synthesis compared with controls. Halothane alone decreased PC synthesis by only 12% and reduced ATP levels by 20%. However, when exposed to both halothane and H2O2 together, ATP levels decreased by 40%, and PC synthesis rates decreased by 51%. Pretreatment of cells with nicotinamide, an inhibitor of poly adenosine diphosphate ribose polymerase, completely prevented the ATP loss and PC synthesis decline caused by H2O2 alone, but it had no effect on the halothane-augmented portion of the cell injury. These data suggest that the ability of halothane to enhance oxidative damage may be related to its own specific effects on cell energetics that may not be amenable to the same treatments used to mitigate other cellular mechanisms of oxidative stress. IMPLICATIONS: A mediator of inflammation (hydrogen peroxide) and an inhaled anesthetic (halothane) interact to decrease cell energy and secretion of a substance (surfactant) required for healthy lung function from cells that line gas-exchange compartments. This interaction represents a possible mechanism by which inflammatory lung disease may become more severe intraoperatively. PMID- 11916802 TI - Intrathecal lidocaine prevents cardiovascular collapse and neurogenic pulmonary edema in a rat model of acute intracranial hypertension. AB - Sympathetic hyperactivity during sudden intracranial hypertension leads to cardiovascular instability, myocardial dysfunction, and neurogenic pulmonary edema. Because spinal anesthesia is associated with sympatholysis, we investigated the protective effects of intrathecal lidocaine in a rodent model. Halothane-anesthetized rats were given a 10-microL intrathecal injection of saline (n = 10) or lidocaine 1% (n = 6). A subdural balloon catheter was inflated for 60 s to produce intracranial hypertension. Hemodynamics were monitored, and hearts and lungs were harvested for histological examination. In Saline versus Lidocaine-Treated rats, peak mean arterial blood pressure during balloon inflation was 115 +/- 4 mm Hg versus 78 +/- 8 mm Hg (P < 0.05), mean arterial blood pressure 30 min after balloon deflation was 47 +/- 2 mm Hg versus 67 +/- 3 mm Hg (P < 0.05), and lung weight was 1.54 +/- 0.03 g versus 1.41 +/- 0.04 g (P < 0.05), respectively. Cardiac dysrhythmias and electrocardiographic changes were more frequent in the Saline-Treated group (P < 0.05). Saline-Treated rats had extensive, hemorrhagic pulmonary edema, whereas the Lidocaine-Treated rats had only patchy areas of lung abnormality. Histological changes in the myocardium were rare, and no difference was found between the two groups. We conclude that intrathecal lidocaine prevents cardiovascular collapse and neurogenic pulmonary edema in a rat model of acute intracranial hypertension. IMPLICATIONS: In a rat model of intracranial balloon inflation, intrathecal lidocaine prevented cardiovascular collapse and neurogenic pulmonary edema. Descending neural pathways are involved in the development of cardiopulmonary complications associated with acute intracranial hypertension. PMID- 11916803 TI - L-arginine and nitroglycerin restore hypercapnia-induced cerebral vasodilation in rabbits after its attenuation by ketamine. AB - Although it has been reported that ketamine attenuates hypercapnia-induced cerebral vasodilation, the mechanism remains unknown. Because nitric oxide is involved in cerebral CO2 reactivity, we studied the effects of L-arginine and nitroglycerin on ketamine-mediated attenuation of vascular responses to hypercapnia. Under pentobarbital anesthesia, 16 rabbits underwent closed cranial window preparation. Hypercapnic challenges were repeated after IV saline, ketamine (10 mg/kg, followed by 20 mg x kg(-1) x h(-1)), or ketamine plus either L-arginine (150 mg/kg, followed by 100 mg x kg(-1) x h(-1); n = 8) or nitroglycerin (5 microg x kg(-1) x min(-1) infusion; n = 8). Ketamine reduced hypercapnia-induced cerebral vasodilation (1.27%/mm Hg +/- 0.45%/mm Hg [saline] versus 0.82%/mm Hg +/- 0.53%/mm Hg [ketamine]: P < 0.05), but L-arginine restored reactivity (1.28%/mm Hg +/- 0.73%/mm Hg: P < 0.05 versus ketamine), as did nitroglycerin (1.14%/mm Hg +/- 0.73%/mm Hg [saline] versus 0.56%/mm Hg +/- 0.63%/mm Hg [ketamine]: P < 0.05, and 1.15%/mm Hg +/- 0.74%/mm Hg [ketamine plus nitroglycerin]: P < 0.05 versus ketamine). This indicates that ketamine attenuates cerebral CO2 reactivity, at least in part, via suppression of nitric oxide-cyclic guanosine monophosphate mechanisms in the cerebral vasculature. IMPLICATIONS: The attenuation of cerebral vasodilation to hypercapnia seen under ketamine anesthesia is reversed by L-arginine or nitroglycerin infusion. PMID- 11916804 TI - Blood patch therapy for spontaneous intracranial hypotension: safe performance after epidurography in an unconscious patient. AB - IMPLICATIONS: Epidurography was useful for identifying the epidural space and determining the likely spread of an epidural blood patch in an unconscious patient with spontaneous intracranial hypotension. PMID- 11916805 TI - The role of cyclooxygenase-1 and -2 in the rat formalin test. AB - Prostaglandins are thought to play an important role in nociceptive transmission at peripheral sites and in the spinal cord. Prostaglandins are produced by cyclooxygenase (COX), which catalyzes the conversion of arachidonic acid. Two forms of COX have been identified: COX-1, which is constitutively expressed, and COX-2, which is an inducible enzyme. To define the role of COX-1 in nociceptive transmission, we examined the effect of oral and intrathecal administration of 5 (4-chlorophenyl)-1-(4-methoxyphenyl)-3-trifluoromethylpyrazole (SC-560), a selective COX-1 inhibitor, on the rat formalin test and compared the effect of SC 560 with that of celecoxib, a COX-2 selective inhibitor, and indomethacin, a nonselective COX-1 and COX-2 inhibitor, on the rat formalin test. Oral and intrathecal administration of SC-560 had no effect on the agitation behavior in the rat formalin test. Oral and intrathecal administration of celecoxib and indomethacin depressed agitation behavior during the rat formalin test. These data suggest that prostaglandins synthesized by COX-1 are not involved in nociceptive transmission during the rat formalin test but that COX-2 does play an important role in the rat formalin test. IMPLICATIONS: Our data suggest that a COX-2 selective inhibitor, but not a COX-1 selective inhibitor, may produce a good analgesic effect on the inflammatory pain state in a clinical situation. PMID- 11916807 TI - The interaction between gabapentin and amitriptyline in the rat formalin test after systemic administration. AB - We examined the effects of systemically administered gabapentin on flinching and biting/licking behaviors produced by 2.5% formalin in the rat, compared these with those of amitriptyline, and determined the effects of combinations of gabapentin with amitriptyline. Gabapentin produced a dose-related inhibition of Phase 2, but not Phase 1, flinching and biting/licking behaviors. In contrast, amitriptyline produced an increase in Phase 2 flinching behaviors while simultaneously decreasing biting/licking behaviors. Fifty percent effective dose (ED50) values against biting/licking behaviors were 22.9 +/- 1.3 mg/kg and 8.5 +/ 1.3 mg/kg for gabapentin and amitriptyline, respectively. Combinations of increasing fractional increments of ED50 doses of gabapentin and amitriptyline produced an additive effect against biting/licking behaviors, as revealed by isobolographic analysis. These increments had no effect on flinching behaviors except at the ED25 + ED25 doses, at which flinching was increased, again revealing additivity between the two drugs. Flinching behaviors in rats do not reflect the analgesic properties of systemically administered amitriptyline observed in humans and may not be useful for predicting an effect of combinations of drugs with amitriptyline. Biting/licking behaviors do reflect analgesic properties for both drugs and may be more useful in this regard. IMPLICATIONS: By use of the rat formalin test, a model of persistent pain, we examined the effect of a combination of amitriptyline and gabapentin, which are used to treat chronic pain in humans. The drug combination produced additive analgesia against one outcome, but another outcome was more ambiguous. PMID- 11916806 TI - Spinal and peripheral mu opioids and the development of secondary tactile allodynia after thermal injury. AB - Local thermal injury to the paw leads to an increased sensitivity to a noxious stimulus applied to the site (primary thermal hyperalgesia) and an increased sensitivity to tactile stimuli in skin sites adjacent to the primary injury (secondary tactile allodynia; 2 degrees TA). We sought to define the peripheral and spinal actions of mu opioids in regulating 2 degrees TA. First, a mild thermal injury was induced on one heel, producing 2 degrees TA. This 2 degrees TA was blocked by pretreatment, but not posttreatment, with a topical mu-opioid agonist, loperamide (1.7%-5%). Second, 2 degrees TA was blocked by intrathecal morphine (0.1-10 microg) pre- and postinjury. 2 degrees TA reappeared when systemic naloxone was given before, but not after, injury in intrathecal morphine pretreated rats. Intrathecal remifentanil, a short-lasting mu-opioid agonist, infused periinjury (3 microg/min), did not block subsequent primary thermal hyperalgesia, but it produced a dose-dependent (0.3-3 microg/min) abolition of 2 degrees TA. Local tissue injury leads to 2 degrees TA by the activation of opiate sensitive afferents and the initiation of a cascade that persists in the absence of that initiating injury-induced stimulus. IMPLICATIONS: Sensitivity to touch observed in areas adjacent to injury is blocked by opioids applied before, but not after, injury. This suggests that injury-activated opioid-sensitive fibers are responsible for sensitization and reveals a cascade that is diminished by pretreatment but not posttreatment, providing a rationale for adequate analgesia before injury (surgery) has occurred. PMID- 11916808 TI - Perioperative small-dose S(+)-ketamine has no incremental beneficial effects on postoperative pain when standard-practice opioid infusions are used. AB - Several studies report that when small-dose racemic ketamine, an N-methyl-D aspartate receptor antagonist, is administered perioperatively, opioid consumption is reduced postoperatively. S(+)-ketamine has a higher affinity for the N-methyl-D-aspartate receptor and less-serious side effects than racemic ketamine. Thirty patients scheduled for elective arthroscopic anterior cruciate ligament repair were enrolled in this randomized, double-blinded clinical trial designed to determine the preemptive effect of S(+)-ketamine on postoperative analgesia requirements in a setting of clinically relevant perioperative analgesia. Total IV anesthesia was induced and maintained with remifentanil (0.125-1.0 microg x kg(-1) x min(-1)) and a propofol target-controlled infusion (target 2-4 microg/mL). The Ketamine group received a bolus of 0.5 mg/kg S(+) ketamine before incision, followed by a continuing infusion of 2 microg x kg(-1) x min(-1) until 2 h after emergence from anesthesia. The Control group received NaCl in the same sequence. After IV morphine provided pain relief down to < or =3 on a visual analog scale scored from 0 to 10, patients were connected to a patient-controlled analgesia device. There were no significant differences between the two groups in terms of total morphine consumption or VAS scores, either at rest or with movement. In our study, S(+)-ketamine did not contribute to postoperative pain reduction, possibly because of the clinically routine perioperative opioid analgesia. IMPLICATIONS: Small-dose S(+)-ketamine had no positive effect on postoperative analgesia when administered perioperatively for elective arthroscopic anterior cruciate ligament repair. Unlike investigations of the racemic mixture of ketamine, our study methods included timely standard practice perioperative opioid analgesia, which seems to make supplemental analgesia unnecessary. PMID- 11916810 TI - The origin of the spinal subdural space: ultrastructure findings. AB - Previous studies of samples from cranial meninges have created doubts about the existence of a virtual subdural space. We examined the ultrastructure of spinal meninges from three human cadavers immediately after death to see whether there is a virtual subdural space at this level. The arachnoid mater had two portions: a compact laminar portion covering the dural sac internal surface and a trabecular portion extending like a spider web around the pia mater. There was a cellular interface between the laminar arachnoid and the internal layer of the dura that we called the dura-arachnoid interface. There was no subdural space in those specimens where the dura mater was macroscopically in continuity with the arachnoid trabecules. In the specimens where the dura mater was separated from the arachnoid, we found fissures in between the neurothelial cells that extended throughout the interface. We hypothesize that the subdural space would have its origin within the dura-arachnoid interface when the neurothelial cells break up, creating in this way a real subdural space. IMPLICATIONS: The subdural space was not seen under transmission electron microscopy in samples of human spinal meninges where surgical manipulation was avoided. Scanning electron microscopy in other samples showed the presence of broken neurothelial cells giving up fissures that extended along the dura-arachnoid interface. These findings may explain the origin of a real subdural space. PMID- 11916809 TI - A double-blinded, randomized comparison of either 0.5% levobupivacaine or 0.5% ropivacaine for sciatic nerve block. AB - To compare intraoperative and postoperative clinical properties of levobupivacaine and ropivacaine for sciatic nerve block, 50 ASA physical status I and II patients undergoing hallux valgus repair received a femoral nerve block with 15 mL of 2% mepivacaine. They were then randomly allocated in a double blinded fashion to receive a sciatic nerve block with either 0.5% levobupivacaine (n = 25) or 0.5% ropivacaine (n = 25). An independent blinded observer evaluated the onset time of surgical anesthesia as well as the quality of the surgical block and postoperative analgesia. The median (range) onset time of surgical block at the sciatic nerve distribution was 30 min (5-60 min) with levobupivacaine and 15 min (5-60 min) with ropivacaine (P = 0.63). Four patients (two patients in each group) received a supplementary ankle block by the surgeon just before the beginning of surgery. All four patients also received IV fentanyl supplementation, but in three of them, propofol infusion was required to complete surgery (two in the Levobupivacaine group [8%] and one in the Ropivacaine group [4%]; P = 0.99). In six patients of the Levobupivacaine group (24%) and five patients of the Ropivacaine group (20%), IV fentanyl supplementation was required to complete surgery (P = 0.99). No differences in the time to recovery of sensory and motor function were observed between the two groups, whereas median (range) duration of postoperative analgesia was 16 h (8-24 h) with levobupivacaine and 16 h (8-24 h) with ropivacaine (P = 0.83). We conclude that 0.5% levobupivacaine and 0.5% ropivacaine provide comparable surgical anesthesia and postoperative analgesia. IMPLICATIONS: No studies have compared the clinical properties of levobupivacaine with those of ropivacaine when providing sciatic nerve block for hallux valgus repair. Results from this prospective, randomized, double-blinded study demonstrate that 20 mL of either 0.5% levobupivacaine or 0.5% ropivacaine provide comparable surgical block with prolonged postoperative analgesia. PMID- 11916811 TI - Postoperative analgesia with continuous sciatic nerve block after foot surgery: a prospective, randomized comparison between the popliteal and subgluteal approaches. AB - To compare the posterior popliteal and subgluteal continuous sciatic nerve block for anesthesia and acute postoperative pain management after foot surgery, 60 ASA physical status I and II patients undergoing elective orthopedic foot surgery were randomly assigned to either a Subgluteal group (n = 30) or Popliteal group (n = 30). Before surgery and after performing a femoral nerve block with 15 mL of 2% mepivacaine, we performed the sciatic nerve block with 20 mL of 0.75% ropivacaine using either a subgluteal or posterior popliteal approach, and the placement of a catheter came afterward. In the recovery room, the catheter was connected to a patient-controlled analgesia pump to infuse 0.2% ropivacaine (basal infusion rate of 5 mL/h, incremental bolus of 10 mL, and a lockout time of 60 min). There were no technical problems in catheter placement. Intraoperative efficacy of nerve block was similar in the two groups. Postoperative catheter displacement and occlusion were recorded in four patients in the Popliteal group and two patients in the Subgluteal group (P = 0.67). Both approaches provided similar postoperative analgesia. We conclude that the subgluteal approach is as effective and safe as the previously described posterior popliteal approach for continuous sciatic block and can be considered a useful alternative to anesthesia and acute postoperative analgesia after foot procedures. IMPLICATIONS: Comparing two different approaches for continuous sciatic nerve block after orthopedic foot surgery, this prospective, randomized study demonstrated that the subgluteal approach is as effective and safe as the previously described posterior popliteal approach, and can be considered a useful alternative to anesthesia and acute postoperative analgesia after foot procedures. PMID- 11916813 TI - Subdural block complicating spinal anesthesia? AB - IMPLICATIONS: Features suggestive of subdural block appeared after an apparently normal subarachnoid block. The long bevel of the reusable Quincke-type spinal needle may have contributed to the development of this complication. We propose that spinal needles should have a smaller bevel to minimize the possibility of such a complication. PMID- 11916812 TI - Continuous three-in-one block for postoperative pain after lower limb orthopedic surgery: where do the catheters go? AB - Continuous three-in-one block is widely used for postoperative analgesia after proximal lower limb surgery, but location of the catheter has not been well addressed in the literature. We prospectively studied, in 100 patients, the characteristics of catheter threading under the iliac fascia and the correlations between catheter tip location and effective sensory and motor blockade of the three principal nerves of the lumbar plexus. Postoperatively, in conscious patients, 16 to 20 cm of a catheter was placed in the fascial sheath after femoral nerve location with a nerve stimulator. Contrast media (3 mL Iopamidol 390) was injected, and the catheter tip was located by means of an anteroposterior pelvic radiograph. An equal-volume mixture of 0.5% bupivacaine/2% lidocaine with epinephrine (30 mL) was injected through the catheter. Patient and catheter-insertion characteristics were noted. Thirty minutes after injection, sensory blockade was evaluated in the cutaneous territories of the lateral femoral cutaneous, femoral, and obturator nerves, along with motor blockade of the last two nerves. Pain scores at 30 min were also recorded. Seven block failures were noted. The tip of the catheter reached the lumbar plexus (Group 1) in 23% of the patients and lay deep to the medial (Group 2) or lateral (Group 3) part of the fascia iliaca in 33% and 37% of the patients, respectively. Demographic data and catheter threading characteristics were comparable among the groups. A three-in-one block was noted in 91% of Group 1 patients, but in only 52% and 27% of Group 2 and 3 patients, respectively (P < 0.05). Comparing Group 2 and 3 patients, sensory block was achieved in respectively 100% and 94% for the femoral nerve, 52% and 94% for the lateral femoral cutaneous nerve (P < 0.05), and 82% and 27% for the obturator nerve (P < 0.05). Visual analog scale pain scores on movement were significantly lower in Group 1 patients (P < 0.05). We conclude that during a continuous three-in-one block, the threaded catheter rarely reached the lumbar plexus. The quality of sensory and motor blockade and initial pain relief depend on the location of the catheter tip under the fascia iliaca. IMPLICATIONS: The course of a continuous three-in-one block catheter is unpredictable. Only 23% of the catheters lie near the lumbar plexus. The success of sensory and motor blocks, as well as postoperative analgesia, depend on the position of the catheter under the fascia iliaca. PMID- 11916814 TI - Possible bupivacaine toxicity after intraarticular injection for postarthroscopic analgesia of the knee: implications of the surgical procedure. AB - IMPLICATIONS: We report a case of possible bupivacaine toxicity after intraarticular injection during knee arthroscopy. The importance of the specific type of surgical procedure performed during arthroscopy and its relationship to potential local anesthetic toxicity are highlighted. PMID- 11916815 TI - Calibrated pneumoperitoneal venting to prevent N2O accumulation in the CO2 pneumoperitoneum during laparoscopy with inhaled anesthesia: an experimental study in pigs. AB - Nitrous oxide (N2O) accumulates in the CO2 pneumoperitoneum during laparoscopy when N2O is used as an adjuvant for inhaled anesthesia. This may worsen the consequences of gas embolism and introduce a fire risk. In this study, we quantified the pneumoperitoneal gas venting necessary to prevent significant contamination by inhaled N2O. Four domestic pigs (26-30 kg) were anesthetized and ventilated with 66% N2O in oxygen. A CO2 pneumoperitoneum was insufflated and maintained at a pressure of 12 mm Hg. Each animal underwent three experimental conditions, in random sequence, for 70 min each: 1) no pneumoperitoneal leak, 2) leak of 2 L every 10 min (12 L/h), and 3) leak of 4 L every 10 min (24 L/h). Every 10 min, pneumoperitoneal gas samples were analyzed for fractions (FPn) of N2O and CO2. Without leaks, FPnN2O increased continually and reached 29.58% +/- 3.15% at 70 min. With leaks of 2 and 4 L every 10 min (12 and 24 L/h), FPnN2O reached a plateau of <10% after 30 min. We conclude that calibrated pneumoperitoneal venting of 12 or 24 L/h is enough to prevent the constitution of potentially dangerous pneumoperitoneal gas mixtures if venting is constant. IMPLICATIONS: External venting calibrated at four or eight initial pneumoperitoneal volumes per hour with compensation by fresh CO2 is sufficient to prevent nitrous oxide buildup of more than 10% in the pneumoperitoneum during laparoscopy with inhaled general anesthesia if venting is constant. PMID- 11916816 TI - The threshold and gain of thermoregulatory vasoconstriction differs during anesthesia in the dependent and upper arms in the lateral position. AB - Increased intraluminal pressure may help maintain vasodilation in a dependent arm even after hypothermia triggers centrally mediated thermoregulatory vasoconstriction. We therefore tested the hypotheses that the threshold (triggering core temperature) and gain (increase in vasoconstriction per degree centigrade) of cold-induced vasoconstriction is reduced in the dependent arm during anesthesia. Anesthesia was maintained with 0.4 minimum alveolar anesthetic concentration of desflurane in 10 volunteers in the left-lateral position. Mean skin temperature was reduced to 31 degrees C to decrease core body temperature. Fingertip blood flow in both arms was measured, as was core body temperature. The vasoconstriction threshold was slightly, but significantly, less in the dependent arm (36.2 degrees C +/- 0.3 degrees C, mean +/- SD) than in the upper arm (36.5 degrees C +/- 0.3 degrees C). However, the gain of vasoconstriction in the dependent arm was 2.3-fold greater than in the upper arm. Consequently, intense vasoconstriction (i.e., a fingertip blood flow of 0.15 mL/min) occurred at similar core temperatures. In the lateral position, the vasoconstriction threshold was reduced in the dependent arm; however, gain was also increased in the dependent arm. The thermoregulatory system may thus recognize that hydrostatic forces reduce the vasoconstriction threshold and may compensate by sufficiently augmenting gain. IMPLICATIONS: The threshold for cold-induced vasoconstriction is reduced in the dependent arm, but the gain of vasoconstriction is increased. Consequently, the core temperature triggering intense vasoconstriction was similar in each arm, suggesting that the thermoregulatory system compensates for the hydrostatic effects of the lateral position. PMID- 11916817 TI - Intubating laryngeal mask airway size selection: a randomized triple crossover study in paralyzed, anesthetized male and female adult patients. AB - We determined the optimal size of intubating laryngeal mask airway (ILM) for ventilation and blind tracheal intubation in men and women. We also determined the distance the tracheal tube needs to protrude beyond the distal aperture to ensure that the cuff is through the vocal cords. Fifty male and 50 female anesthetized, paralyzed patients (ASA physical status I or II, aged 18-80 yr) were studied. Three operators (A, B, and C) were involved for the purposes of blinding. The size 3, 4, or 5 ILM was inserted into each patient in random order by Operator A, and the quality of ventilation was scored (adequate, suboptimal, or failed) by Operator B. The fiberoptic position (correct, too shallow, or too deep) and the distance between the distal aperture and the vocal cords was determined by Operator B. A single attempt at blind intubation was made by Operator C. Operators B and C were blinded to the size of the ILM. Operator C was also blinded to the information recorded by Operator B. All ILMs were inserted into the laryngopharynx at the first attempt. For men and women, the ventilation score was smaller for the Size 3 than the Size 4 or 5 (all: P < 0.002). For men, correct positioning was less common with the Size 3 than the Size 4 or 5 (both: P < 0.02). For women, correct positioning was similar among sizes. For men, tracheal intubation was successful less frequently with the Size 3 (84%) than the Size 4 (100%) or 5 (98%) (both: P < or = 0.01). For women, tracheal intubation success was similar among sizes (Size 3, 4, and 5: 86%, 96%, and 92%, respectively). Intubation was always successful if the ILM was correctly positioned and always failed if it was too shallow or deep. In both male and female patients, the distance between the distal aperture and the vocal cords increased with increasing ILM size (all: P < 0.04) and patient height (P < 0.0001) and was always longer for men (all: P < 0.0001). The overall mean distance (95% confidence interval) that the tracheal tube needed to protrude was 10-12 cm (8-13 cm) in men and 8-11 cm (8-12 cm) in women. We conclude that for men, the Size 4 and 5 ILMs are better than the Size 3 for ventilation and blind intubation. For women, the Size 4 and 5 ILMs are better than the Size 3 for ventilation, but there is no difference among sizes for blind intubation. The length the tracheal tube must protrude from the distal aperture to ensure that the cuff is completely through the vocal cords is 8-13 cm, depending on ILM size, the tracheal tube size, and the sex and height of the patient. IMPLICATIONS: For men, the Size 4 and 5 intubating laryngeal mask airways are better than the Size 3 for ventilation and blind tracheal intubation. For women, the Size 4 and 5 are better than the Size 3 for ventilation, but there is no difference among sizes for blind intubation. The length the tracheal tube must protrude from the distal aperture of the intubating laryngeal mask airway to ensure that the cuff is completely through the vocal cords is 8-13 cm, depending on the size of the mask and tracheal tube and on the sex and height of the patient. PMID- 11916819 TI - Acute postoperative glaucoma after nonocular surgery remains a diagnostic challenge. AB - IMPLICATIONS: Anesthesia may acutely reveal angle-closure glaucoma. This complication is an ophthalmologic emergency. However, symptoms of acute glaucoma may be overlooked or misinterpreted in a sedated or comatose patient, and this may result in delayed treatment. Immediate diagnosis and appropriate treatment should be done to prevent visual loss. PMID- 11916818 TI - The differential effect of halothane and 1,2-dichlorohexafluorocyclobutane on in vitro muscle contractures of patients susceptible to malignant hyperthermia. AB - Malignant hyperthermia (MH) is an autosomal dominant, potentially fatal pharmacogenetic disorder of skeletal muscle. Approximately half of all known MH families show a linkage to the ryanodine receptor type 1 (RY1) gene. Although our knowledge of the diagnosis, genetics, and therapy of MH has improved, the exact pathogenesis and the role of volatile anesthetics as trigger substances for an MH crisis remain unknown. Compounds that do not obey the Meyer-Overton hypothesis (i.e., nonimmobilizers) are today an important part of research on anesthetic mechanisms. We designed this study to test the hypothesis that the nonimmobilizer 1,2-dichlorohexafluorocyclobutane (2N) compared with halothane has different effects on in vitro muscle contractures of muscle bundles from MH-susceptible (MHS) individuals. In vitro muscle contracture tests were performed with either halothane (approximately 660 microM, equivalent to approximately 4 minimum alveolar anesthetic concentration [MAC]) or 2N ( approximately 100 microM, equivalent to approximately 5 times predicted MAC). MAC is defined as the anesthetic concentration that prevents nocifensive movements after a surgical stimulus in 50% of subjects. In contrast to halothane, 2N caused only minimal muscle contractures in muscle bundles from six MHS patients (0.13 g [0.04-0.31 g] vs 1.95 g [1.60-4.70 g], median values and ranges; P = 0.004). Halothane and 2N differ in their effects on muscle contractures of MHS individuals, possibly because of a differing action on MH RY1. IMPLICATIONS: Using in vitro contracture tests, we showed that halothane and the nonimmobilizer 1,2 dichlorohexafluorocyclobutane differ in their effects on contractures of muscle bundles from individuals susceptible to malignant hyperthermia (MH) as a result of their differing action on MH ryanodine receptors. These findings render this receptor a possible molecular target for volatile anesthetic action. PMID- 11916820 TI - Esophageal insufflation with normal fiberoptic positioning of the ProSeal laryngeal mask airway. AB - IMPLICATIONS: Esophageal insufflation can occur simultaneously with venting from the drain tube during positive pressure ventilation with the ProSeal laryngeal mask airway. PMID- 11916821 TI - Reduction of pain on injection of propofol: a comparison of fentanyl with remifentanil. PMID- 11916822 TI - When is an airway not an airway? PMID- 11916823 TI - Searching the preferred anesthetic technique during one-lung-ventilation. PMID- 11916824 TI - Is 0.375% bupivacaine safe in caudal anesthesia in neonates and young infants? PMID- 11916825 TI - Hiccupping and regurgitation via the drain tube of the ProSeal laryngeal mask. PMID- 11916826 TI - Propofol formulation and pain on injection. PMID- 11916827 TI - Diagnosis of diastolic dysfunction: importance of spectral Doppler imaging. PMID- 11916828 TI - Warming blankets should not be placed over transdermal medications. PMID- 11916829 TI - Propofol-induced bronchoconstriction: asthma or allergy? PMID- 11916831 TI - Simulation of ultrasound backscattering by red cell aggregates: effect of shear rate and anisotropy. AB - Tissue characterization using ultrasound (US) scattering allows extraction of relevant cellular biophysical information noninvasively. Characterization of the level of red blood cell (RBC) aggregation is one of the proposed application. In the current paper, it is hypothesized that the microstructure of the RBCs is a main determinant of the US backscattered power. A simulation model was developed to study the effect of various RBC configurations on the backscattered power. It is an iterative dynamical model that considers the effect of the adhesive and repulsive forces between RBCs, and the effect of the flow. The method is shown to be efficient to model polydispersity in size, shape, and orientation of the aggregates due to the flow, and to relate these variations to the US backscattering properties. Three levels of aggregability at shear rates varying between 0.05 and 10 s(-1) were modeled at 40% hematocrit. The simulated backscattered power increased with a decrease in the shear rate or an increase in the RBC aggregability. Angular dependence of the backscattered power was observed. It is the first attempt to model the US power backscattered by RBC aggregates polydisperse in size and shape due to the shearing of the flow. PMID- 11916830 TI - Characterization of the azide-dependent bacteriorhodopsin-like photocycle of salinarum halorhodopsin. AB - The photocycle of salinarum halorhodopsin was investigated in the presence of azide. The azide binds to the halorhodopsin with 150 mM binding constant in the absence of chloride and with 250 mM binding constant in the presence of 1 M chloride. We demonstrate that the azide-binding site is different from that of chloride, and the influence of chloride on the binding constant is indirect. The analysis of the absorption kinetic signals indicates the existence of two parallel photocycles. One belongs to the 13-cis retinal containing protein and contains a single red shifted intermediate. The other photocycle, of the all trans retinal containing halorhodopsin, resembles the cycle of bacteriorhodopsin and contains a long-living M intermediate. With time-resolved spectroscopy, the spectra of intermediates were determined. Intermediates L, N, and O were not detected. The multiexponential rise and decay of the M intermediate could be explained by the introduction of the "spectrally silent" intermediates M1, M2, and HR', HR, respectively. The electric signal measurements revealed the existence of a component equivalent with a proton motion toward the extracellular side of the membrane, which appears during the M1 to M2 transition. The differences between the azide-dependent photocycle of salinarum halorhodopsin and pharaonis halorhodopsin are discussed. PMID- 11916832 TI - The role of dimerization in prion replication. AB - The central theme in prion diseases is the conformational transition of a cellular protein from a physiologic to a pathologic (so-called scrapie) state. Currently, two alternative models exist for the mechanism of this autocatalytic process; in the template assistance model the prion is assumed to be a monomer of the scrapie conformer, whereas in the nucleated polymerization model it is thought to be an amyloid rod. A recent variation on the latter assumes disulfide reshuffling as the mechanism of polymerization. The existence of stable dimers, let alone their mechanistic role, is not taken into account in either of these models. In this paper we review evidence supporting that the dimerization of either the normal or the scrapie state, or both, has a decisive role in prion replication. The contribution of redox changes, i.e., the temporary opening and possible rearrangement of the intramolecular disulfide bridge is also considered. We present a model including these features largely ignored so far and show that it adheres satisfactorily to the observed phenomenology of prion replication. PMID- 11916833 TI - Segregation of photosystems in thylakoid membranes as a critical phenomenon. AB - The distribution of the two photosystems, PSI and PSII, in grana and stroma lamellae of the chloroplast membranes is not uniform. PSII are mainly concentrated in grana and PSI in stroma thylakoids. The dynamics and factors controlling the spatial segregation of PSI and PSII are generally not well understood, and here we address the segregation of photosystems in thylakoid membranes by means of a molecular dynamics method. The lateral segregation of photosystems was studied assuming a model comprising a two-dimensional (in plane), two-component, many-body system with periodic boundary conditions and competing interactions between the photosystems in the thylakoid membrane. PSI and PSII are represented by particles with different values of negative charge. The pair interactions between particles include a screened Coulomb repulsive part and an exponentially decaying attractive part. The modeling results suggest a complicated phase behavior of the system, including quasi-crystalline phase of randomly distributed complexes of PSII and PSI at low ionic screening, well defined clustered state of segregated complexes at high screening, and in addition, an intermediate agglomerate phase where the photosystems tend to aggregate together without segregation. The calculations demonstrated that the ordering of photosystems within the membrane was the result of interplay between electrostatic and lipid-mediated interactions. At some values of the model parameters the segregation can be represented visually as well as by analyzing the correlation functions of the configuration. PMID- 11916834 TI - Simulations of zwitterionic and anionic phospholipid monolayers. AB - Results of atomistic molecular dynamics simulations of dipalmitoylphosphatidylcholine and dipalmitoylphosphatidylglycerol monolayers at the air/water interface are presented. Dipalmitoylphosphatidylcholine is zwitterionic and dipalmitoylphosphatidylglycerol is anionic at physiological pH. NaCl and CaCl2 water subphases are simulated. The simulations are carried out at different surface densities, and a simulation cell geometry is chosen that greatly facilitates the investigation of phospholipid monolayer properties. Ensemble average monolayer properties calculated from simulation are in agreement with experimental measurements. The dependence of the properties of the monolayers on the surface density, the type of the headgroup, and the ionic environment are explained in terms of atomistically detailed pair distribution functions and electron density profiles, demonstrating the strength of simulations in investigating complex, multicomponent systems of biological importance. PMID- 11916835 TI - Effective rate models for receptors distributed in a layer above a surface: application to cells and Biacore. AB - In the Biacore biosensor, a widely used tool for studying the kinetics of ligand/receptor binding, receptors are commonly localized to the sensor surface through attachment to polymers that extend from the surface to form a layer. The importance of the polymeric layer in analyzing data is controversial. The question of the effect of a binding layer also arises in the case of ligands interacting with binding sites distributed in the extracellular matrix of cells. To identify and quantify the effects of a binding layer on the estimation of association and dissociation rate constants, we derived effective rate coefficients. The expressions show that rate constants determined under the standard assumption that binding takes place on a two-dimensional surface underestimate the true reaction rate constants by a factor that depends on the ratio of the height of the layer to the mean free path of the ligand within the layer. We show that, for typical biological ligands, receptors, cells, and Biacore conditions, the binding layer will affect the interpretation of data only if transport of the ligand in the layer is slowed substantially--by one or two orders of magnitude--relative to transport outside the layer. From existing experiments and theory, it is not clear which Biacore experiments, if any, have transport within the dextran layer reduced to such an extent. We propose a method, based on the effective rate coefficients we have derived, for the experimental determination of ligand diffusion coefficients in a polymeric matrix. PMID- 11916837 TI - Electrostatics explains the shift in VDAC gating with salt activity gradient. AB - We have analyzed voltage-dependent anion-selective channel (VDAC) gating on the assumption that the states occupied by the channel are determined mainly by their electrostatic energy. The voltage dependence of VDAC gating both in the presence and in the absence of a salt activity gradient was explained just by invoking electrostatic interactions. A model describing this energy in the main VDAC states has been developed. On the basis of the model, we have considered how external factors cause the redistribution of the channels among their conformational states. We propose that there is a difference in the electrostatic interaction between the voltage sensor and fixed charge within the channel when the former is located in the cis side of membrane as opposed to the trans. This could be the main cause of the shift in the probability curve. The theory describes satisfactorily the experimental data (Zizi et al., Biophys. J. 1998. 75:704-713) and explains some peculiarities of VDAC gating. The asymmetry of the probability curve was related to the apparent location of the VDAC voltage sensor in the open state. By analyzing published experimental data, we concluded that this apparent location is influenced by the diffusion potential. Also discussed is the possibility that VDAC gating at high voltage may be better described by assuming that the mobile charge consists of two parts that have to overcome different energetic barriers in the channel-closing process. PMID- 11916836 TI - Echinocyte shapes: bending, stretching, and shear determine spicule shape and spacing. AB - We study the shapes of human red blood cells using continuum mechanics. In particular, we model the crenated, echinocytic shapes and show how they may arise from a competition between the bending energy of the plasma membrane and the stretching/shear elastic energies of the membrane skeleton. In contrast to earlier work, we calculate spicule shapes exactly by solving the equations of continuum mechanics subject to appropriate boundary conditions. A simple scaling analysis of this competition reveals an elastic length Lambda(el), which sets the length scale for the spicules and is, thus, related to the number of spicules experimentally observed on the fully developed echinocyte. PMID- 11916838 TI - Forces required of kinesin during processive transport through cytoplasm. AB - The purpose of this paper is to deduce whether the maximum force, steplike movement, and rate of ATP consumption of kinesin, as measured in buffer, are sufficient for the task of fast transport of vesicles in cells. Our results show that moving a 200-nm vesicle in viscoelastic COS7 cytoplasm, with the same steps as observed for kinesin-driven beads in buffer, required a maximum force of 16 pN and work per step of 1 +/- 0.7 ATP, if the drag force was assumed to decrease to zero between steps. In buffer, kinesin can develop a force of 6-7 pN while consuming 1 ATP/step, comparable to the required values. As an alternative to assuming that the force vanishes between steps, the measured COS7 viscoelasticity was extrapolated to zero frequency by a numerical fit. The force required to move the bead then exceeded 75 pN at all times and peaked briefly to 92 pN, well beyond the measured capabilities of a single kinesin in buffer. The work per step increased to 7 +/- 5 ATP, greatly exceeding the energy available to a single motor. PMID- 11916839 TI - Long time dynamics of Met-enkephalin: comparison of explicit and implicit solvent models. AB - Met-enkephalin is one of the smallest opiate peptides. Yet, its dynamical structure and receptor docking mechanism are still not well understood. The conformational dynamics of this neuron peptide in liquid water are studied here by using all-atom molecular dynamics (MD) and implicit water Langevin dynamics (LD) simulations with AMBER potential functions and the three-site transferable intermolecular potential (TIP3P) model for water. To achieve the same simulation length in physical time, the full MD simulations require 200 times as much CPU time as the implicit water LD simulations. The solvent hydrophobicity and dielectric behavior are treated in the implicit solvent LD simulations by using a macroscopic solvation potential, a single dielectric constant, and atomic friction coefficients computed using the accessible surface area method with the TIP3P model water viscosity as determined here from MD simulations for pure TIP3P water. Both the local and the global dynamics obtained from the implicit solvent LD simulations agree very well with those from the explicit solvent MD simulations. The simulations provide insights into the conformational restrictions that are associated with the bioactivity of the opiate peptide dermorphin for the delta-receptor. PMID- 11916840 TI - Binding and diffusion of CheR molecules within a cluster of membrane receptors. AB - Adaptation of the attractant response in Escherichia coli is attributable to the methylation of its transmembrane chemotactic receptors by the methyltransferase CheR. This protein contains two binding domains, one for the sites of methylation themselves and the other for a flexible tether at the C terminus of the receptor. We have explored the theoretical consequences of this binding geometry for a CheR molecule associated with a cluster of chemotactic receptors. Calculations show that the CheR molecule will bind with high net affinity to the receptor lattice, having a high probability of being attached by one or both of its domains at any instant of time. Because of the relatively low affinity of its individual domains and the close proximity of neighboring receptors, it is likely that when one domain unbinds it will reattach to the array before the other domain unbinds. Stochastic simulations show that the enzyme will move through the receptor cluster in a hand-over-hand fashion, like a gibbon swinging through the branches of a tree. We explore the possible consequences of this motion, which we term "molecular brachiation", for chemotactic adaptation and suggest that a similar mechanism may be operative in other large assemblies of protein molecules. PMID- 11916841 TI - Molecular dynamics simulation of dipalmitoylphosphatidylserine bilayer with Na+ counterions. AB - We performed a molecular dynamics simulation of dipalmitoylphosphatidylserine (DPPS) bilayer with Na+ counterions. We found that hydrogen bonding between the NH group and the phosphate group leads to a reduction in the area per headgroup when compared to the area in dipalmitoylphosphatidylcholine bilayer. The Na+ ions bind to the oxygen in the carboxyl group of serine, thus giving rise to a dipolar bilayer similar to dipalmitoylphosphatidylethanolamine bilayer. The results of the simulation show that counterions play a crucial role in determining the structural and electrostatic properties of DPPS bilayer. PMID- 11916842 TI - Measurement of the lateral diffusion of human MHC class I molecules on HeLa cells by fluorescence recovery after photobleaching using a phycoerythrin probe. AB - The mobility of cell surface MHC class I molecules on HeLa cells was measured by fluorescence recovery after photobleaching (FRAP). The probe used for these studies was the phycobiliprotein R-phycoerythrin coupled to Fab fragments of a monoclonal antibody specific for human monomorphic MHC class I molecules. It was found that the recovery curves could be equally well fitted by either a random diffusion model with an immobile component or by an anomalous diffusion model. In the latter case, the anomalous diffusion exponent was consistent with that previously determined by single-particle tracking (SPT) experiments using the same probe (P. R. Smith, I. E. G. Morrison, K. M. Wilson, N. Fernandez, and R. J. Cherry. 1999. Biophys. J. 76:3331-3344). The FRAP experiments, however, yielded a considerably higher value of D(0), the diffusion coefficient for a time interval of 1 s. To determine whether the results were probe dependent, FRAP measurements were also performed with the same monoclonal antibody labeled with Oregon Green. These experiments gave similar results to those obtained with the phycoerythrin probe. FRAP experiments with the lipid probe 5-N-(octadecanoyl) aminofluoroscein (ODAF) bound to HeLa cells gave typical results for lipid diffusion. Overall, our observations and analysis are consistent with anomalous diffusion of MHC class I diffusion on HeLa cells, but quantitative differences between FRAP and SPT data remain to be explained. PMID- 11916843 TI - Comparison of PSGL-1 microbead and neutrophil rolling: microvillus elongation stabilizes P-selectin bond clusters. AB - A cell-scaled microbead system was used to analyze the force-dependent kinetics of P-selectin adhesive bonds independent of micromechanical properties of the neutrophil's surface microvilli, an elastic structure on which P-selectin ligand glycoprotein-1 (PSGL-1) is localized. Microvillus extension has been hypothesized in contributing to the dynamic range of leukocyte rolling observed in vivo during inflammatory processes. To evaluate PSGL-1/P-selectin bond kinetics of microbeads and neutrophils, rolling and tethering on P-selectin-coated substrates were compared in a parallel-plate flow chamber. The dissociation rates for PSGL-1 microbeads on P-selectin were briefer than those of neutrophils for any wall shear stress, and increased more rapidly with increasing flow. The microvillus length necessary to reconcile dissociation constants of PSGL-1 microbeads and neutrophils on P-selectin was 0.21 microm at 0.4 dyn/cm2, and increased to 1.58 microm at 2 dyn/cm2. The apparent elastic spring constant of the microvillus ranged from 1340 to 152 pN/microm at 0.4 and 2.0 dyn/cm2 wall shear stress. Scanning electron micrographs of neutrophils rolling on P-selectin confirmed the existence of micrometer-scaled tethers. Fixation of neutrophils to abrogate microvillus elasticity resulted in rolling behavior similar to PSGL-1 microbeads. Our results suggest that microvillus extension during transient PSGL-1/P-selectin bonding may enhance the robustness of neutrophil rolling interactions. PMID- 11916844 TI - Initial stages of cell-matrix adhesion can be mediated and modulated by cell surface hyaluronan. AB - A conceptual temporal and spatial gap exists between the first encounter of a cell with an adhesive substrate and the advanced stages of focal adhesion formation. Although ample information is available on focal adhesions structure and function, the mechanism of the first interaction events and the nature of the molecules mediating them are largely unknown. In this paper we identify cell surface-associated hyaluronan as a mediator and modulator of the first steps of adhesion of A6 and other cells to conventional tissue culture substrates as well as to the surfaces of calcium-(R,R)-tartrate tetrahydrate crystals. Treatment of A6 cells with hyaluronidase suppresses their rapid interactions with these adhesive substrates, and incubation of either the hyaluronidase-treated cells or the substrate with hyaluronan restores cell adhesion. In contrast, excess hyaluronan on both the cells and the substrate strongly inhibits adhesion. We thus propose that cell-surface-associated hyaluronan can mediate and modulate cell-matrix adhesion at the very first encounter with the substrate. It may promote it through the establishment of exquisitely stereospecific chemical interactions or inhibit it by virtue of steric exclusion and/or electrostatic repulsion. PMID- 11916845 TI - Kinetics and mechanism of intercellular ice propagation in a micropatterned tissue construct. AB - Understanding the effects of cell-cell interaction on intracellular ice formation (IIF) is required to design optimized protocols for cryopreservation of tissue. To determine the effects of cell-cell interactions during tissue freezing, without confounding effects from uncontrolled factors (such as time in culture, cell geometry, and cell-substrate interactions), HepG2 cells were cultured in pairs on glass coverslips micropatterned with polyethylene glycol disilane, such that each cell interacted with exactly one adjacent cell. Assuming the cell pair to be a finite state system, being either in an unfrozen state (no ice in either cell), a singlet state (IIF in one cell only), or a doublet state (IIF in both cells), the kinetics of state transitions were theoretically modeled and cryomicroscopically measured. The rate of intercellular ice propagation, estimated from the measured singlet state probability, increased in the first 24 h of culture and remained steady thereafter. In cell pairs cultured for 24 h and treated with the gap junction blocker 18beta-glycyrrhetinic acid before freezing, the intercellular ice propagation rate was lower than in untreated controls (p < 0.001), but significantly greater than zero (p < 0.0001). These results suggest that gap junctions mediate some, but not all, mechanisms of ice propagation in tissue. PMID- 11916847 TI - Modeling of the pore domain of the GLUR1 channel: homology with K+ channel and binding of channel blockers. AB - Molecular models of the M2 segments of the GluR1 channel have been elaborated using a molecular mechanics approach. The models are based on the homology between pore-lining segments of AMPA receptor channels and the KcsA K+ channel and on cyclic H bonds at the Q/R site of the AMPA receptor channel. The N terminal region of an M2 segment of the channel is assumed, like that of the K+ channel, to adopt a helical conformation. Due to a deletion, the C-terminal end of the M2 segment of the AMPA receptor is more stretched than that of the K+ channel. As a result, only a single oxygen ring may be exposed to the AMPA receptor channel pore. Data on the block of AMPA receptor channels by dicationic adamantane derivatives have been used to select the most relevant model. The model with the oxygen of a Gly residue (position +2 from the Q/R site) exposed to the pore best fits the experimental data. This model also fits experimental data for another class of AMPA receptor antagonists, the polyamine amides. According to the model, the side-chains of the C-terminal residues are involved in intra receptor interactions that stabilize the structure of the channel rather than in interactions with ions in the pore. PMID- 11916846 TI - MYO1A (brush border myosin I) dynamics in the brush border of LLC-PK1-CL4 cells. AB - The kidney epithelial cell line, LLC-PK1-CL4 (CL4), forms a well ordered brush border (BB) on its apical surface. CL4 cells were used to examine the dynamics of MYO1A (M1A; formerly BB myosin I) within the BB using GFP-tagged MIA (GFP-M1A), MIA motor domain (GFP-MDIQ), and tail domain (GFP-Tail). GFP-beta-actin (GFP Actin) was used to assess actin dynamics within the BB. GFP-M1A, GFP-Tail, but not GFP-MDIQ localized to the BB, indicating that the tail is sufficient for apical targeting of M1A. GFP-Actin targeted to all the actin domains of the cell including the BB. Fluorescence recovery after photobleaching analysis revealed that GFP-M1A and GFP-Tail turnover in the BB is rapid, approximately 80% complete in <1 min. As expected for an actin-based motor, ATP depletion resulted in significant inhibition of GFP-M1A turnover yet had little effect on GFP-Tail exchange. Rapid turnover of GFP-M1A and GFP-Tail was not due to actin turnover as GFP-Actin turnover in the BB was much slower. These results indicate that the BB population of M1A turns over rapidly, while its head and tail domains interact transiently with the core actin and plasma membrane, respectively. This rapidly exchanging pool of M1A envelops an actin core bundle that, by comparison, is static in structure. PMID- 11916848 TI - Mechanism of inactivation gating of human T-type (low-voltage activated) calcium channels. AB - Recovery from inactivation of T-type Ca channels is slow and saturates at moderate hyperpolarizing voltage steps compared with Na channels. To explore this unique kinetic pattern we measured gating and ionic currents in two closely related isoforms of T-type Ca channels. Gating current recovers from inactivation much faster than ionic current, and recovery from inactivation is much more voltage dependent for gating current than for ionic current. There is a lag in the onset of gating current recovery at -80 mV, but no lag is discernible at -120 mV. The delay in recovery from inactivation of ionic current is much more evident at all voltages. The time constant for the decay of off gating current is very similar to the time constant of deactivation of open channels (ionic tail current), and both are strongly voltage dependent over a wide voltage range. Apparently, the development of inactivation has little influence on the initial deactivation step. These results suggest that movement of gating charge occurs for inactivated states very quickly. In contrast, the transitions from inactivated to available states are orders of magnitude slower, not voltage dependent, and are rate limiting for ionic recovery. These findings support a deactivation-first path for T-type Ca channel recovery from inactivation. We have integrated these concepts into an eight-state kinetic model, which can account for the major characteristics of T-type Ca channel inactivation. PMID- 11916850 TI - Subunit-selective contribution to channel gating of the M4 domain of the nicotinic receptor. AB - The muscle nicotinic receptor (AChR) is a pentamer of four different subunits, each of which contains four transmembrane domains (M1-M4). We recently showed that channel opening and closing rates of the AChR depend on a hydrogen bond involving a threonine at position 14' of the M4 domain in the alpha-subunit. To determine whether residues in equivalent positions in non-alpha-subunits contribute to channel gating, we mutated deltaT14', betaT14', and epsilonS14' and evaluated changes in the kinetics of acetylcholine-activated currents. The mutation epsilonS14'A profoundly slows the rate of channel closing, an effect opposite to that produced by mutation of alphaT14'. Unlike mutations of alphaT14', epsilonS14'A does not affect the rate of channel opening. Mutations in deltaT14' and betaT14' do not affect channel opening or closing kinetics, showing that conserved residues are not functionally equivalent in all subunits. Whereas alphaT14'A and epsilonS14'A subunits contribute additively to the closing rate, they contribute nonadditively to the opening rate. Substitution of residues preserving the hydrogen bonding ability at position 14' produce nearly normal gating kinetics. Thus, we identify subunit-specific contributions to channel gating of equivalent residues in M4 and elucidate the underlying mechanistic and structural bases. PMID- 11916849 TI - Protein kinase C phosphorylation of purified Na,K-ATPase: C-terminal phosphorylation sites at the alpha- and gamma-subunits close to the inner face of the plasma membrane. AB - The alpha-subunit of the Na,K-ATPase is phosphorylated at specific sites by protein kinases A and C. Phosphorylation by protein kinase C (PKC) is restricted to the N terminus and takes place to a low stoichiometry, except in rat. Here we show that the alpha-subunit of shark Na,K-ATPase can be phosphorylated by PKC at C-terminal sites to stoichiometric levels in the presence of detergents. Two novel phosphorylation sites are possible candidates for this PKC phosphorylation: Thr-938 in the M8/M9 loop located very close to the PKA site, and Ser-774, in the proximal part of the M5/M6 hairpin. Both sites are highly conserved in all known alpha-subunits, indicating a physiological role. A similar pattern of detergent mediated phosphorylation by PKC was found in pig kidney Na,K-ATPase alpha subunit. Interestingly, the kidney-specific gamma-subunit was phosphorylated by PKC in the presence of detergent. The close proximity of the novel PKC sites to the membrane suggests that targeting proteins to tether PKC into the membrane phase is important in controlling the in vivo phosphorylation of this novel class of membrane-adjacent PKC sites. It is suggested that in purified preparations where functional targeting may be impaired detergents are needed to expose the sites. PMID- 11916851 TI - Applying hidden Markov models to the analysis of single ion channel activity. AB - Hidden Markov models have recently been used to model single ion channel currents as recorded with the patch clamp technique from cell membranes. The estimation of hidden Markov models parameters using the forward-backward and Baum-Welch algorithms can be performed at signal to noise ratios that are too low for conventional single channel kinetic analysis; however, the application of these algorithms relies on the assumptions that the background noise be white and that the underlying state transitions occur at discrete times. To address these issues, we present an "H-noise" algorithm that accounts for correlated background noise and the randomness of sampling relative to transitions. We also discuss three issues that arise in the practical application of the algorithm in analyzing single channel data. First, we describe a digital inverse filter that removes the effects of the analog antialiasing filter and yields a sharp frequency roll-off. This enhances the performance while reducing the computational intensity of the algorithm. Second, the data may be contaminated with baseline drifts or deterministic interferences such as 60-Hz pickup. We propose an extension of previous results to consider baseline drift. Finally, we describe the extension of the algorithm to multiple data sets. PMID- 11916852 TI - Stoichiometry of the Cardiac Na+/Ca2+ exchanger NCX1.1 measured in transfected HEK cells. AB - The stoichiometry with which the Na+/Ca2+ exchanger, NCX1, binds and transports Na+ and Ca2+ has dramatic consequences for ionic homeostasis and cellular function of heart mycocytes and brain neurons, where the exchanger is highly expressed. Previous studies have examined this question using native NCX1 in its endogenous environment. We describe here whole-cell voltage clamp studies using recombinant rat heart NCX1.1 expressed heterologously in HEK-293 cells. This system provides the advantages of a high level of NCX1 protein expression, very low background ion transport levels, and excellent control over clamped voltage and ionic composition. Using ionic conditions that allowed bi-directional currents, voltage ramps were employed to determine the reversal potential for NCX1.1-mediated currents. Analysis of the relation between reversal potential and external [Na+] or [Ca2+], under a variety of intracellular conditions, yielded coupling ratios for Na+ of 1.9-2.3 ions per net charge and for Ca2+ of 0.45 +/- 0.03 ions per net charge. These data are consistent with a stoichiometry for the NCX1.1 protein of 4 Na+ to 1 Ca2+ to 2 charges moved per transport cycle. PMID- 11916854 TI - Ryanodine-induced structural alterations in the RyR channel suggested by neomycin block. AB - In Mead and Williams, (Biophys. J. 82:1953-1963, 2002) we have reported that neomycin is a potent partial blocker of single purified sheep cardiac SR calcium release channels. Neomycin is unusual in that it is capable of blocking when applied to either the cytosolic or the luminal face of the channel. Block at either aspect of the channel is both concentration- and voltage-dependent, but exhibits different blocking parameters. In this study we have investigated the actions of neomycin on ion handling in the ryanodine-modified channel. Neomycin is more effective at the cytosolic face, having a Kb(0) value of 534.9 +/- 35.17 nM compared with a Kb(0) value of 971.5 +/- 66.62 nM for the luminal face. The voltage dependence also differs at the two sites. Values of zdelta for cytosolic and luminal neomycin are 1.09 +/- 0.04 and -0.57 +/- 0.03, respectively. The interaction of neomycin with the ryanodine-modified channel differs notably from that in the unmodified channel. Voltage-dependent relief of block is not observed after ryanodine modification, and the luminal blocking characteristics are altered. This suggests that ryanodine induces changes at the luminal mouth of the channel and may confer increased rigidity to the channel protein. PMID- 11916853 TI - Block of the ryanodine receptor channel by neomycin is relieved at high holding potentials. AB - In this study we have investigated the actions of the aminoglycoside antibiotic neomycin on K+ conductance in the purified sheep cardiac sarcoplasmic reticulum (SR) calcium-release channel (RyR). Neomycin induces a concentration- and voltage dependent partial block from both the cytosolic and luminal faces of the channel. Blocking parameters for cytosolic and luminal block are markedly different. Neomycin has a greater affinity for the luminal site of interaction than the cytosolic site: zero-voltage dissociation constants (Kb(0)) are respectively 210.20 +/- 22.80 and 589.70 +/- 184.00 nM for luminal and cytosolic block. However, neomycin also exhibits voltage-dependent relief of block at holding potentials >+60 mV when applied to the cytosolic face and a similar phenomenon may occur with luminal neomycin at high negative holding potentials. These observations indicate that, under appropriate conditions, neomycin is capable of passing through the RyR channel. PMID- 11916855 TI - Reservoir boundaries in Brownian dynamics simulations of ion channels. AB - Brownian dynamics (BD) simulations provide a practical method for the calculation of ion channel conductance from a given structure. There has been much debate about the implementation of reservoir boundaries in BD simulations in recent years, with claims that the use of improper boundaries could have large effects on the calculated conductance values. Here we compare the simple stochastic boundary that we have been using in our BD simulations with the recently proposed grand canonical Monte Carlo method. We also compare different methods of creating transmembrane potentials. Our results confirm that the treatment of the reservoir boundaries is mostly irrelevant to the conductance properties of an ion channel as long as the reservoirs are large enough. PMID- 11916856 TI - Syringomycin E channel: a lipidic pore stabilized by lipopeptide? AB - Highly reproducible ion channels of the lipopeptide antibiotic syringomycin E demonstrate unprecedented involvement of the host bilayer lipids. We find that in addition to a pronounced influence of lipid species on the open-channel ionic conductance, the membrane lipids play a crucial role in channel gating. The effective gating charge, which characterizes sensitivity of the conformational equilibrium of the syringomycin E channels to the transmembrane voltage, is modified by the lipid charge and lipid dipolar moment. We show that the type of host lipid determines not only the absolute value but also the sign of the gating charge. With negatively charged bilayers, the gating charge sign inverts with increased salt concentration or decreased pH. We also demonstrate that the replacement of lamellar lipid by nonlamellar with the negative spontaneous curvature inhibits channel formation. These observations suggest that the asymmetric channel directly incorporates lipids. The charges and dipoles resulting from the structural inclusion of lipids are important determinants of the overall energetics that underlies channel gating. We conclude that the syringomycin E channel may serve as a biophysical model to link studies of ion channels with those of lipidic pores in membrane fusion. PMID- 11916857 TI - Functional characterization of the type 1 inositol 1,4,5-trisphosphate receptor coupling domain SII(+/-) splice variants and the Opisthotonos mutant form. AB - The type 1 inositol (1,4,5)-trisphosphate receptor (InsP3R1) plays a critical role in Ca2+ signaling in cells. Neuronal and nonneuronal isoforms of the InsP3R1 differ by alternative splicing in the coupling domain of the InsP3R1 (SII site). Deletion of 107 amino acids from the coupling domain of the InsP3R1 results in epileptic-like behaviors in opisthotonos (opt) spontaneous mouse mutant. Using Spodoptera frugiperda cells expression system, we compared single-channel behavior of recombinant InsP3R1-SII(+), InsP3R1-SII(-), and InsP3R1-opt channels in planar lipid bilayers. The main results of our study are: 1) the InsP3R1-SII( ) has a higher conductance (94 pS) and the InsP3R1-opt has a lower conductance (64 pS) than the InsP3R1-SII(+) (81 pS); 2) the bell-shaped Ca2+-dependence peaks at 200-300 nM Ca2+ for all three InsP3R1 isoforms; 3) the bell-shaped Ca2+ dependence is wider for the InsP3R1-SII(+) and narrower for the InsP3R1-SII(-) and InsP3R1-opt; 4) the apparent affinity for ATP is sixfold lower for the InsP3R1-SII(-) (1.4 mM) and 20-fold lower for the InsP3R1-opt (5.3 mM) than for the InsP3R1-SII(+) (0.24 mM); 5) the InsP3R1-SII(-) is approximately twofold more active than the InsP3R1-SII(+) in the absence of ATP. Obtained results provide novel information about the molecular determinants of the InsP3R1 function. PMID- 11916858 TI - Ca2+-activated K+ currents regulate odor adaptation by modulating spike encoding of olfactory receptor cells. AB - The olfactory system is thought to accomplish odor adaptation through the ciliary transduction machinery in olfactory receptor cells (ORCs). However, ORCs that have lost their cilia can exhibit spike frequency accommodation in which the action potential frequency decreases with time despite a steady depolarizing stimulus. This raises the possibility that somatic ionic channels in ORCs might serve for odor adaptation at the level of spike encoding, because spiking responses in ORCs encode the odor information. Here I investigate the adaptational mechanism at the somatic membrane using conventional and dynamic patch-clamp recording techniques, which enable the ciliary mechanism to be bypassed. A conditioning stimulus with an odorant-induced current markedly shifted the response range of action potentials induced by the same test stimulus to higher concentrations of the odorant, indicating odor adaptation. This effect was inhibited by charybdotoxin and iberiotoxin, Ca2+-activated K+ channel blockers, suggesting that somatic Ca2+-activated K+ currents regulate odor adaptation by modulating spike encoding. I conclude that not only the ciliary machinery but also the somatic membrane currents are crucial to odor adaptation. PMID- 11916860 TI - Fluctuation of surface charge in membrane pores. AB - Surface charge in track-etched polyethylene terephthalate (PET) membranes with narrow pores has been probed with a fluorescent cationic dye (3,3' diethyloxacarbocyanine iodide (diO-C2-(3))) using confocal microscopy. Staining of negatively charged PET membranes with diO-C2-(3) is a useful measure of surface charge for the following reasons: 1) the dye inhibits K(+) currents through the pores and reduces their selectivity for cations; 2) it inhibits [3H] choline+ transport and promotes 36Cl- transport across the membrane in a pH- and ionic-strength-dependent fashion; and 3) staining of pores by diO-C2-(3) is reduced by low pH and by the presence of divalent cations such as Ca2+ and Zn2+. Measurement of the time dependence of cyanine staining of pores shows fluctuations of fluorescence intensity that occur on the same time scale as do fluctuations of ionic current in such pores. These data support our earlier proposal that fluctuations in ionic current across pores in synthetic and biological membranes reflect fluctuations in the surface charge of the pore walls in addition to molecular changes in pore proteins. PMID- 11916859 TI - Hemichannel and junctional properties of connexin 50. AB - Lens fiber connexins, cx50 and cx46 (alpha3 and alpha8), belong to a small subset of connexins that can form functional hemichannels in nonjunctional membranes. Knockout of either cx50 or cx46 results in a cataract, so the properties of both connexins are likely essential for proper physiological functioning of the lens. Although portions of the sequences of these two connexins are nearly identical, their hemichannel properties are quite different. Cx50 hemichannels are much more sensitive to extracellular acidification than cx46 hemichannels and differ from cx46 hemichannels both in steady-state and kinetic properties. Comparison of the two branches of the cx50 hemichannel G-V curve with the junctional G-V curve suggests that cx50 gap junctions gate with positive relative polarity. The histidine-modifying reagent, diethyl pyrocarbonate, reversibly blocks cx50 hemichannel currents but not cx46 hemichannel currents. Because cx46 and cx50 have very similar amino acid sequences, one might expect that replacing the two histidines unique to the third transmembrane region of cx50 with the corresponding cx46 residues would produce mutants more closely resembling cx46. In fact this does not happen. Instead the mutant cx50H161N does not form detectable hemichannels but forms gap junctions indistinguishable from wild type. Cx50H176Q is oocyte lethal, and the double mutant, cx50H61N/H176Q, neither forms hemichannels nor kills oocytes. PMID- 11916861 TI - Segregation of saturated chain lipids in pulmonary surfactant films and bilayers. AB - The physical properties of organized system (bilayers and monolayers at the air water interface) composed of bovine lipid extract surfactant (BLES) were studied using correlated experimental techniques. 6-Dodecanoyl-2-dimethylamino naphthalene (LAURDAN)-labeled giant unilamelar vesicles (mean diameter approximately 30 microm) composed of BLES were observed at different temperatures using two-photon fluorescence microscopy. As the temperature was decreased, dark domains (gel-like) appeared at physiological temperature (37 degrees C) on the surface of BLES giant unilamelar vesicles. The LAURDAN two-photon fluorescent images show that the gel-like domains span the lipid bilayer. Quantitative analysis of the LAURDAN generalized polarization function suggests the presence of a gel/fluid phase coexistence between 37 degrees C to 20 degrees C with low compositional and energetic differences between the coexisting phases. Interestingly, the microscopic scenario of the phase coexistence observed below 20 degrees C shows different domain's shape compared with that observed between 37 degrees C to 20 degrees C, suggesting the coexistence of two ordered but differently organized lipid phases on the bilayer. Epifluorescence microscopy studies of BLES monomolecular films doped with small amounts of fluorescent lipids showed the appearance and growth of dark domains (liquid condensed) dispersed in a fluorescent phase (liquid expanded) with shapes and sizes similar to those observed in BLES giant unilamelar vesicles. Our study suggests that bovine surfactant lipids can organize into discrete phases in monolayers or bilayers with equivalent temperature dependencies and may occur at physiological temperatures and surface pressures equivalent to those at the lung interface. PMID- 11916862 TI - A computer simulation of functional group contributions to free energy in water and a DPPC lipid bilayer. AB - A series of all-atom molecular dynamics simulations has been performed to evaluate the contributions of various functional groups to the free energy of solvation in water and a dipalmitoylphospatidylcholine lipid bilayer membrane and to the free energies of solute transfer (Delta(DeltaG(o))X) from water into the ordered-chain interior of the bilayer. Free energies for mutations of the alpha-H atom in p-toluic acid to six different substituents (-CH3, -Cl, -OCH3, -CN, -OH, COOH) were calculated by a combined thermodynamic integration and perturbation method and compared to literature results from vapor pressure measurements, partition coefficients, and membrane transport experiments. Convergence of the calculated free energies was indicated by substantial declines in standard deviations for the calculated free energies with increased simulation length, by the independence of the ensemble-averaged Boltzmann factors to simulation length, and the weak dependence of hysteresis effects on simulation length over two different simulation lengths and starting from different initial configurations. Calculated values of Delta(DeltaG(o))X correlate linearly with corresponding values obtained from lipid bilayer transport experiments with a slope of 1.1 and from measurements of partition coefficients between water and hexadecane or decadiene, with slopes of 1.1 and 0.9, respectively. Van der Waals interactions between the functional group of interest and the acyl chains in the ordered chain region account for more than 95% of the overall potential energy of interaction. These results support the view that the ordered chain region within the bilayer interior is the barrier domain for transport and that solvation interactions within this region resemble those occurring in a nonpolar hydrocarbon. PMID- 11916864 TI - Agarose-dextran gels as synthetic analogs of glomerular basement membrane: water permeability. AB - Novel agarose-dextran hydrogels were synthesized and their suitability as experimental models of glomerular basement membrane was examined by measuring their Darcy (hydraulic) permeabilities (kappa). Immobilization of large dextran molecules in agarose was achieved by electron beam irradiation. Composite gels were made with agarose volume fractions (phi(a)) of 0.04 or 0.08 and dextran volume fractions (phi(d)) ranging from 0 to 0.02 (fiber volume/gel volume), using either of two dextran molecular weights (500 or 2000). At either agarose concentration and for either size of dextran, kappa decreased markedly as the amount of dextran was increased. Statistically significant deviations from the value of kappa for pure agarose were obtained for remarkably small volume fractions of dextran: phi(d) > or = 0.0003 for phi(a) = 0.04 and phi(d) > or = 0.001 for phi(a) = 0.08. The Darcy permeabilities were much more sensitive to phi(d) than to phi(a), and were as much as 26 times smaller than those of pure agarose. Although phi(d) was an important variable, dextran molecular weight was not. The effects of dextran addition on kappa were described fairly well using simple structural idealizations. At high agarose concentrations, the dextran chains behaved as fine fibers interspersed among coarse agarose fibrils, whereas, at low concentrations, the dextran molecules began to resemble spherical obstacles embedded in agarose gels. The ability to achieve physiologically relevant Darcy permeabilities with these materials (as low as 1.6 nm2) makes them an attractive experimental model for glomerular basement membrane and possibly other extracellular matrices. PMID- 11916863 TI - Conformational characterization of ceramides by nuclear magnetic resonance spectroscopy. AB - Ceramide (Cer) has been identified as an active lipid second messenger in the regulation of cell growth, differentiation, and apoptosis. Its analog, dihydroceramide, without the 4 to 5 trans double bond in the sphingoid backbone lacks these biological effects. To establish the conformational features that distinguish ceramide from its analogs, nuclear magnetic resonance spectral data were acquired for diluted samples of ceramides (C2- and C18-Cer), dihydroceramide (C16-DHCer), and deoxydihydroceramide (C18-DODHCer). Our results suggest that in both C2- and C18-Cer, an H-bond network is formed in which the amide proton NH is donated to the OH groups on carbons C1 and C3 of the sphingosine backbone. Two tightly bound water molecules appear to stabilize this network by participating in flip-flop interactions with the hydroxyl groups. In DHCer, the lack of the trans double bond leads to a conformational distortion of this H-bonding motif. Without the critical double bond, the degree with which water molecules stabilize the H bonds between the two OH groups of the sphingolipid is reduced. This structural alteration might preclude the participation of DHCer in signaling related interactions with cellular targets. PMID- 11916865 TI - Osmotic and curvature stress affect PEG-induced fusion of lipid vesicles but not mixing of their lipids. AB - Poly (ethylene glycol) (PEG) in the external environment of membrane vesicles creates osmotic imbalance that leads to mechanical stress in membranes and may induce local membrane curvature. To determine the relative importance of membrane stress and curvature in promoting fusion, we monitored contents mixing (CM) and lipid mixing (LM) between different sized vesicles under a variety of osmotic conditions. CM between highly curved vesicles (SUV, 26 nm diameter) was up to 10 times greater than between less curved vesicles (LUV, 120 nm diameter) after 5 min incubation at a low PEG concentration (<10 wt%), whereas LM was only approximately 30% higher. Cryo-electron microscopy showed that PEG at 10 wt% did not create high curvature contacts between membranes in LUV aggregates. A negative osmotic gradient (-300 mOs/kg, hypotonic inside) increased CM two- to threefold for both types of vesicles, but did not affect LM. A positive gradient (+220 mOs/kg, hypertonic inside) nearly eliminated CM and had no effect on LM. Hexadecane added to vesicles had no effect on LM but enhanced CM and reduced the inhibitory effect on CM of a positive osmotic gradient, but had little influence on results obtained under a negative osmotic gradient. We conclude that the ability of closely juxtaposed bilayers to form an initial intermediate ("stalk") as soon as they come into close contact was not influenced by osmotic stress or membrane curvature, although pore formation was critically dependent on these stresses. The results also suggest that hexadecane affects the same part of the fusion process as osmotic stress. We interpret this result to suggest that both a negative osmotic gradient and hexadecane reduce the unfavorable free energy of hydrophobic interstices associated with the intermediates of the fusion process. PMID- 11916866 TI - The depth of porphyrin in a membrane and the membrane's physical properties affect the photosensitizing efficiency. AB - Photosensitized biological processes, as applied in photodynamic therapy, are based on light-triggered generation of molecular singlet oxygen by a membrane residing sensitizer. Most of the sensitizers currently used are hydrophobic or amphiphilic porphyrins and their analogs. The possible activity of the short lived singlet oxygen is limited to the time it is diffusing in the membrane, before it emerges into the aqueous environment. In this paper we demonstrate the enhancement of the photosensitization process that is obtained by newly synthesized protoporphyrin derivatives, which insert their tetrapyrrole chromophore deeper into the lipid bilayer of liposomes. The insertion was measured by fluorescence quenching by iodide and the photosensitization efficiency was measured with 9,10-dimethylanthracene, a fluorescent chemical target for singlet oxygen. We also show that when the bilayer undergoes a melting phase transition, or when it is fluidized by benzyl alcohol, the sensitization efficiency decreases because of the enhanced diffusion of singlet oxygen. The addition of cholesterol or of dimyristoyl phosphatydilcholine to the bilayer moves the porphyrin deeper into the bilayer; however, the ensuing effect on the sensitization efficiency is different in these two cases. These results could possibly define an additional criterion for the choice and design of hydrophobic, membrane-bound photosensitizers. PMID- 11916867 TI - Structural characterization of weakly attached cross-bridges in the A*M*ATP state in permeabilized rabbit psoas muscle. AB - It is well established that in a skeletal muscle under relaxing conditions, cross bridges exist in a mixture of four weak binding states in equilibrium (A*M*ATP, A*M*ADP*P(i), M*ATP, and M*ADP*P(i)). It has been shown that these four weak binding states are in the pathway to force generation. In the past their structural, biochemical, and mechanical properties have been characterized as a group. However, it was shown that the myosin heads in the M*ATP state exhibited a disordered distribution along the thick filament, while in the M*ADP*P(i) state they were well ordered. It follows that the structures of the weakly attached states of A*M*ATP and A*M*ADP*P(i) could well be different. Individual structures of the two attached states could not be assigned because protocol for isolating the two states has not been available until recently. In the present study, muscle fibers are reacted with N-phenylmaleimide such that ATP hydrolysis is inhibited, i.e., the cross-bridge population under relaxing conditions is distributed only between the two states of M*ATP and A*M*ATP. Two-dimensional x ray diffraction was applied to determine the structural characteristics of the attached A*M*ATP state. Because the detached state of M*ATP is disordered and does not contribute to layer line intensities, changes as a result of increasing attachment in the A*M*ATP state are attributable to that state alone. The equilibrium toward the attached state was achieved by lowering the ionic strength. The results show that upon attachment, both the myosin and the first actin associated layer lines increased intensities, while the sixth actin layer line was not significantly affected. However, the intensities remain weak despite substantial attachment. The results, together with modeling (see J. Gu, S. Xu and L. C. Yu, 2002, Biophys. J. 82:2123-2133), suggest that there is a wide range of orientation of the attached A*M*ATP cross-bridges while the myosin heads maintain some degree of helical distribution on the thick filament, suggesting a high degree of flexibility in the actomyosin complex. Furthermore, the lack of sensitivity of the sixth actin layer line suggests that the binding site on actin differs from the putative site for rigor binding. The significance of the flexibility in the A*M*ATP complex in the process of force generation is discussed. PMID- 11916868 TI - A model of cross-bridge attachment to actin in the A*M*ATP state based on x-ray diffraction from permeabilized rabbit psoas muscle. AB - A model of cross-bridges binding to actin in the weak binding A*M*ATP state is presented. The modeling was based on the x-ray diffraction patterns from the relaxed skinned rabbit psoas muscle fibers where ATP hydrolysis was inhibited by N-phenylmaleimide treatment (S. Xu, J. Gu, G. Melvin, L. C. Yu. 2002. Biophys. J. 82:2111-2122). Calculations included both the myosin filaments and the actin filaments of the muscle cells, and the binding to actin was assumed to be single headed. To achieve a good fit, considerable flexibility in the orientation of the myosin head and the position of the S1-S2 junction is necessary, such that the myosin head can bind to a nearby actin whereas the tail end was kept in the proximity of the helical track of the myosin filament. Hence, the best-fit model shows that the head binds to actin in a wide range of orientations, and the tail end deviates substantially from its lattice position in the radial direction (approximately 60 A). Surprisingly, the best fit model reveals that the detached head, whose location thus far has remained undetected, seems to be located close to the surface of the myosin filament. Another significant requirement of the best-fit model is that the binding site on actin is near the N terminus of the actin subunit, a position distinct from the putative rigor-binding site. The results support the idea that the essential role played by the weak binding states M*ATP <--> A*M*ATP for force generation lies in its flexibility, because the probability of attachment is greatly increased, compared with the weak binding M*ADP*P(i) <--> A*M*ADP*P(i) states. PMID- 11916870 TI - Kinetic evidence for three photolyzable taxonomic conformational substates in oxymyoglobin. AB - The kinetics of oxygen geminate binding with the taxonomic substates of MbO2 are reported. The maximum entropy method was used to analyze the rebinding kinetics of MbCO and MbO2 monitored in the Soret. The resulting rate distributions were found to consist of a small number of overlapping bands. A global parametric fit of a series of rate distributions recorded at several temperatures was performed using a Gaussian basis set to resolve the individual enthalpy distributions P(H). This approach was first validated by showing that the well-documented taxonomic substates of MbCO could be recovered. The method was then applied to MbO2. Three taxonomic substates were identified at pH 4.8, whereas only two of them contribute to oxygen geminate rebinding at pH 7.0. These findings show that, similarly to MbCO, MbO2 also exists as three photolyzable and kinetically different taxonomic substates and suggest reconsidering the issue of the photolysis quantum yield of MbO2. PMID- 11916869 TI - The biochemical kinetics underlying actin movement generated by one and many skeletal muscle myosin molecules. AB - To better understand how skeletal muscle myosin molecules move actin filaments, we determine the motion-generating biochemistry of a single myosin molecule and study how it scales with the motion-generating biochemistry of an ensemble of myosin molecules. First, by measuring the effects of various ligands (ATP, ADP, and P(i)) on event lifetimes, tau(on), in a laser trap, we determine the biochemical kinetics underlying the stepwise movement of an actin filament generated by a single myosin molecule. Next, by measuring the effects of these same ligands on actin velocities, V, in an in vitro motility assay, we determine the biochemistry underlying the continuous movement of an actin filament generated by an ensemble of myosin molecules. The observed effects of P(i) on single molecule mechanochemistry indicate that motion generation by a single myosin molecule is closely associated with actin-induced P(i) dissociation. We obtain additional evidence for this relationship by measuring changes in single molecule mechanochemistry caused by a smooth muscle HMM mutation that results in a reduced P(i)-release rate. In contrast, we observe that motion generation by an ensemble of myosin molecules is limited by ATP-induced actin dissociation (i.e., V varies as 1/tau(on)) at low [ATP], but deviates from this relationship at high [ATP]. The single-molecule data uniquely provide a direct measure of the fundamental mechanochemistry of the actomyosin ATPase reaction under a minimal load and serve as a clear basis for a model of ensemble motility in which actin attached myosin molecules impose a load. PMID- 11916871 TI - Probing the proton channel and the retinal binding site of Natronobacterium pharaonis sensory rhodopsin II. AB - The sensory rhodopsin II from Natronobacterium pharaonis (NpSRII) was mutated to try to create functional properties characteristic of bacteriorhodopsin (BR), the proton pump from Halobacterium salinarum. Key residues from the cytoplasmic and extracellular proton transfer channel of BR as well as from the retinal binding site were chosen. The single site mutants L40T, F86D, P183E, and T204A did not display altered function as determined by the kinetics of their photocycles. However, the photocycle of each of the subsequent multisite mutations L40T/F86D, L40T/F86D/P183E, and L40T/F86D/P183E/T204A was quite different from that of the wild-type protein. The reprotonation of the Schiff base could be accelerated approximately 300- to 400-fold, to approximately two to three times faster than the corresponding reaction in BR. The greatest effect is observed for the quadruple mutant in which Thr-204 is replaced by Ala. This result indicates that mutations affecting conformational changes of the protein might be of decisive importance for the creation of BR-like functional properties. PMID- 11916872 TI - Interaction of proteins in solution from small-angle scattering: a perturbative approach. AB - In this work an improved methodology for studying interactions of proteins in solution by small-angle scattering is presented. Unlike the most common approach, where the protein-protein correlation functions g(ij)(r) are approximated by their zero-density limit (i.e., the Boltzmann factor), we propose a more accurate representation of g(ij)(r) that takes into account terms up to the first order in the density expansion of the mean-force potential. This improvement is expected to be particularly effective in the case of strong protein-protein interactions at intermediate concentrations. The method is applied to analyze small-angle x ray scattering data obtained as a function of the ionic strength (from 7 to 507 mM) from acidic solutions of beta-lactoglobulin at the fixed concentration of 10 gl(-1). The results are compared with those obtained using the zero-density approximation and show significant improvement, particularly in the more demanding case of low ionic strength. PMID- 11916874 TI - Absorption and CD spectroscopy and modeling of various LH2 complexes from purple bacteria. AB - The absorption (OD) and circular dichroism (CD) spectra of LH2 complexes from various purple bacteria have been measured and modeled. Based on the lineshapes of the spectra we can sort the LH2 complexes into two distinguishable groups: "acidophila"-like (type 1) and "molischianum"-like (type 2). Starting from the known geometric structures of Rhodopseudomonas (Rps.) acidophila and Rhodospirillum (Rsp.) molischianum we can model the OD and CD spectra of all species by just slightly varying some key parameters: the interaction strength, the energy difference of alpha- and beta-bound B850 bacteriochlorophylls (BChls), the orientation of the B800 and B850 BChls, and the (in)homogeneous broadening. Although the ring size can vary, the data are consistent with all the LH2 complexes having basically very similar structures. PMID- 11916873 TI - Deuterium/hydrogen exchange factors measured by solution nuclear magnetic resonance spectroscopy as indicators of the structure and topology of membrane proteins. AB - Deuterium/hydrogen exchange factors (chi) were measured for the backbone amide sites of the membrane-bound forms of the 50-residue fd coat protein and the 23 residue magainin2 peptide in lipid micelles by solution nuclear magnetic resonance spectroscopy. By combining kinetic and thermodynamic effects, deuterium/hydrogen exchange factors overcome the principal limitations encountered in the measurements of kinetic protection factors and thermodynamic fractionation factors for membrane proteins. The magnitudes of the exchange factors can be correlated with the structure and topology of membrane-associated polypeptides. In fd coat protein, residues in the transmembrane helix have exchange factors that are substantially smaller than those in the amphipathic surface helix or the loop connecting the two helices. For the amphipathic helical peptide, magainin2, the exchange factors of residues exposed to the solvent are appreciably larger than those that face the hydrocarbon portion of membrane bilayers. These examples demonstrate that deuterium/hydrogen exchange factors can be measured by solution NMR spectroscopy and used to identify residues in transmembrane helices as well as to determine the polarity of amphipathic helices in membrane proteins. PMID- 11916875 TI - Investigating structural changes induced by nucleotide binding to RecA using difference FTIR. AB - Nucleotide binding to RecA results in either the high-DNA affinity form (Adenosine 5'-triphosphate (ATP)-bound) or the more inactive protein conformation associated with a lower affinity for DNA (Adenosine 5'-diphosphate (ADP)-bound). Many of the key structural differences between the RecA-ATP and RecA-ADP bound forms have yet to be elucidated. We have used caged-nucleotides and difference FTIR in efforts to obtain a comprehensive understanding of the molecular changes induced by nucleotide binding to RecA. The photochemical release of nucleotides (ADP and ATP) from biologically inactive precursors was used to initiate nucleotide binding to RecA. Here we present ATP hydrolysis assays and fluorescence studies suggesting that the caged nucleotides do not interact with RecA before photochemical release. Furthermore, we now compare difference spectra obtained in H2O and D2O as our first attempt at identifying the origin of the vibrations influenced by nucleotide binding. The infrared data suggest that unique alpha-helical, beta structures, and side chain rearrangements are associated with the high- and low-DNA affinity forms of RecA. Difference spectra obtained over time isolate contributions arising from perturbations in the nucleotide phosphates and have provided further information about the protein structural changes involved in nucleotide binding and the allosteric regulation of RecA. PMID- 11916876 TI - Three-dimensional cellular deformation analysis with a two-photon magnetic manipulator workstation. AB - The ability to apply quantifiable mechanical stresses at the microscopic scale is critical for studying cellular responses to mechanical forces. This necessitates the use of force transducers that can apply precisely controlled forces to cells while monitoring the responses noninvasively. This paper describes the development of a micromanipulation workstation integrating two-photon, three dimensional imaging with a high-force, uniform-gradient magnetic manipulator. The uniform-gradient magnetic field applies nearly uniform forces to a large cell population, permitting statistical quantification of select molecular responses to mechanical stresses. The magnetic transducer design is capable of exerting over 200 pN of force on 4.5-microm-diameter paramagnetic particles and over 800 pN on 5.0-microm ferromagnetic particles. These forces vary within +/-10% over an area 500 x 500 microm2. The compatibility with the use of high numerical aperture (approximately 1.0) objectives is an integral part of the workstation design allowing submicron-resolution, three-dimensional, two-photon imaging. Three dimensional analyses of cellular deformation under localized mechanical strain are reported. These measurements indicate that the response of cells to large focal stresses may contain three-dimensional global deformations and show the suitability of this workstation to further studying cellular response to mechanical stresses. PMID- 11916877 TI - Stress response in Caenorhabditis elegans caused by optical tweezers: wavelength, power, and time dependence. AB - Optical tweezers have emerged as a powerful technique for micromanipulation of living cells. Although the technique often has been claimed to be nonintrusive, evidence has appeared that this is not always the case. This work presents evidence that near-infrared continuous-wave laser light from optical tweezers can produce stress in Caenorhabditis elegans. A transgenic strain of C. elegans, carrying an integrated heat-shock-responsive reporter gene, has been exposed to laser light under a variety of illumination conditions. It was found that gene expression was most often induced by light of 760 nm, and least by 810 nm. The stress response increased with laser power and irradiation time. At 810 nm, significant gene expression could be observed at 360 mW of illumination, which is more than one order of magnitude above that normally used in optical tweezers. In the 700-760-nm range, the results show that the stress response is caused by photochemical processes, whereas at 810 nm, it mainly has a photothermal origin. These results give further evidence that the 700-760-nm wavelength region is unsuitable for optical tweezers and suggest that work at 810 nm at normal laser powers does not cause stress at the cellular level. PMID- 11916878 TI - A two-photon view of an enzyme at work: Crotalus atrox venom PLA2 interaction with single-lipid and mixed-lipid giant unilamellar vesicles. AB - We describe the interaction of Crotalus atrox-secreted phospholipase A2 (sPLA2) with giant unilamellar vesicles (GUVs) composed of single and binary phospholipid mixtures visualized through two-photon excitation fluorescent microscopy. The GUV lipid compositions that we examined included 1-palmitoyl-2-oleoyl phosphatidylcholine, 1,2-dipalmitoyl-sn-glycero-3-phosphocholine (DPPC), and 1,2 dimyristoyl-sn-glycero-3-phosphocholine (DMPC) (above their gel-liquid crystal transition temperatures) and two well characterized lipid mixtures, 1,2 dimyristoyl-sn-glycero-3-phosphoethanolamine (DMPE):DMPC (7:3) and 1,2-dilauroyl sn-glycero-3-phosphocholine (DLPC)/1,2-diarachidoyl-sn-glycero-3-phosphocholine (DAPC) (1:1) equilibrated at their phase-coexistence temperature regime. The membrane fluorescence probes, 6-lauroyl-2-(dimethylamino) napthalene, 6-propionyl 2-(dimethylamino) naphthalene, and rhodamine-phosphatidylethanolamine, were used to assess the state of the membrane and specifically mark the phospholipid domains. Independent of their lipid composition, all GUVs were reduced in size as sPLA2-dependent lipid hydrolysis proceeded. The binding of sPLA2 was monitored using a fluorescein-sPLA2 conjugate. The sPLA2 was observed to associate with the entire surface of the liquid phase in the single phospholipid GUVs. In the mixed lipid GUV's, at temperatures promoting domain coexistence, a preferential binding of the enzyme to the liquid regions was also found. The lipid phase of the GUV protein binding region was verified by the introduction of 6-propionyl-2 (dimethylamino) naphthalene, which partitions quickly into the lipid fluid phase. Preferential hydrolysis of the liquid domains supported the conclusions based on the binding studies. sPLA2 hydrolyzes the liquid domains in the binary lipid mixtures DLPC:DAPC and DMPC:DMPE, indicating that the solid-phase packing of DAPC and DMPE interferes with sPLA2 binding, irrespective of the phospholipid headgroup. These studies emphasize the importance of lateral packing of the lipids in C. atrox sPLA2 enzymatic hydrolysis of a membrane surface. PMID- 11916879 TI - Analysis of kinetics using a hybrid maximum-entropy/nonlinear-least-squares method: application to protein folding. AB - A hybrid analysis that combines the maximum entropy method (MEM) with nonlinear least squares (NLS) fitting has been developed to interpret a general time dependent signal. Data that include processes of opposite sign and a slow baseline drift can be inverted to obtain both a continuous distribution of lifetimes and a sum of discrete exponentials. Fits by discrete exponentials are performed with initial parameters determined from the distribution of lifetimes obtained with the MEM. The regularization of the parameter space achieved by the MEM stabilizes the introduction of each successive exponential in the NLS fits. This hybrid approach is particularly useful when fitting by a large number of exponentials. Revision of the MEM "prior" based on features in the data can improve the lifetime distribution obtained. Standard errors in the mean are estimated automatically for raw data. The results presented for simulated data and for fluorescence measurements of protein folding illustrate the utility and accuracy of the hybrid algorithm. Analysis of the folding of dihydrofolate reductase reveals six kinetic processes, one more than previously reported. PMID- 11916880 TI - Cellular organization and substructure measured using angle-resolved low coherence interferometry. AB - We measure the organization and substructure of HT29 epithelial cells in a monolayer using angle-resolved low-coherence interferometry. This new technique probes cellular structure by measuring scattered light, as in flow cytometry, but offers an advantage in that the structure can be examined in situ, avoiding the need to disrupt the cell monolayer. We determine the size distribution of the cell nuclei by fitting measured light-scattering spectra to the predictions of Mie theory. In addition, we obtain information about the cellular organization and substructure by examining the spatial correlations within the monolayer. A remarkable finding is that the spatial correlations over small length scales take the form of an inverse power law, indicating the fractal nature of the packing of the subcellular structures. We also identify spatial correlations on a scale large compared with the size of a cell, indicating an overlying order within the monolayer. PMID- 11916882 TI - "The unfriendly games". PMID- 11916881 TI - A new deformation model of hard alpha-keratin fibers at the nanometer scale: implications for hard alpha-keratin intermediate filament mechanical properties. AB - The mechanical behavior of human hair fibers is determined by the interactions between keratin proteins structured into microfibrils (hard alpha-keratin intermediate filaments), a protein sulfur-rich matrix (intermediate filaments associated proteins), and water molecules. The structure of the microfibril matrix assembly has already been fully characterized using electron microscopy and small-angle x-ray scattering on unstressed fibers. However, these results give only a static image of this assembly. To observe and characterize the deformation of the microfibrils and of the matrix, we have carried out time resolved small-angle x-ray microdiffraction experiments on human hair fibers stretched at 45% relative humidity and in water. Three structural parameters were monitored and quantified: the 6.7-nm meridian arc, which is related to an axial separation between groups of molecules along the microfibrils, the microfibril's radius, and the packing distance between microfibrils. Using a surface lattice model of the microfibril, we have described its deformation as a combination of a sliding process and a molecular stretching process. The radial contraction of the matrix is also emphasized, reinforcing the hydrophilic gel nature hypothesis. PMID- 11916883 TI - Do drug cheats ever prosper? PMID- 11916884 TI - To review or not to review, that is the question. PMID- 11916885 TI - "Overuse"--an overused term? PMID- 11916886 TI - Management of the sprained ankle. PMID- 11916887 TI - Mass casualties and triage at a sporting event. AB - When a sports medicine doctor provides coverage for a sporting event with a large number of athletes and spectators, he or she should always be aware that the potential for a large number of injuries exists. In the event of a mass casualty incident that overwhelms the available medical resources, he or she may be the most qualified professional present to triage and organise patient care. Certain basic rules of triage in a disaster situation should be followed, the goal being to save as many lives as possible. Special circumstances, such as crush injuries, lightning strikes, and blast injuries, may affect the triage and initial care of injured patients. PMID- 11916888 TI - Methods, advantages, and limitations of body cooling for exercise performance. AB - Precooling studies confirm that increasing body heat is a limiting factor during exercise. However, it seems that precooling is only beneficial for endurance exercise of up to 30-40 minutes rather than intermittent or short duration exercise. PMID- 11916890 TI - Bioenergetic constraints on tactical decision making in middle distance running. AB - BACKGROUND: The highest velocity that a runner can sustain during middle distance races is defined by the intersection of the runner's individual velocity-time curve and the distance-time curve. The velocity-time curve is presumably fixed at the onset of a race; however, whereas the race distance is ostensibly fixed, the actual distance-time curve is not. That is, it is possible for a runner to run further than the race distance if he or she runs wide on bends in track races. In this instance, the point of intersection of the individual velocity-time curve and the distance-time curve will move downwards and to the right, reducing the best average velocity that can be sustained for the distance. METHODS: To illustrate this point, the race tactics used by the gold and silver medallists at 800 m and 5000 m in the Sydney Olympics were analysed. The paths taken by the runners were carefully tracked and the total distance they covered during the races and the average velocity they sustained over the distances they actually covered were calculated. RESULTS: In both the Olympic 800 m and 5000 m finals, for example, the winner was not the runner who ran at the highest average velocity in the race. Rather, the winners of these races were able to husband their metabolic resources to better effect by running closer to the actual race distance. CONCLUSIONS: Race results in middle distance running events are dependent not just on the energetic potential of the runners at the start of the race and their strategy for pace allocation, but also on the effect of their tactical approach to positioning on the total distance covered in the race. Middle distance runners should be conscious of minimising the distance covered in races if they wish to optimise their performance. PMID- 11916889 TI - A retrospective case-control analysis of 2002 running injuries. AB - OBJECTIVE: To provide an extensive and up to date database for specific running related injuries, across the sexes, as seen at a primary care sports medicine facility, and to assess the relative risk for individual injuries based on investigation of selected risk factors. METHODS: Patient data were recorded by doctors at the Allan McGavin Sports Medicine Centre over a two year period. They included assessment of anthropometric, training, and biomechanical information. A model was constructed (with odds ratios and their 95% confidence intervals) of possible contributing factors using a dependent variable of runners with a specific injury and comparing them with a control group of runners who experienced a different injury. Variables included in the model were: height, weight, body mass index, age, activity history, weekly activity, history of injury, and calibre of runner. RESULTS: Most of the study group were women (54%). Some injuries occurred with a significantly higher frequency in one sex. Being less than 34 years old was reported as a risk factor across the sexes for patellofemoral pain syndrome, and in men for iliotibial band friction syndrome, patellar tendinopathy, and tibial stress syndrome. Being active for less than 8.5 years was positively associated with injury in both sexes for tibial stress syndrome; and women with a body mass index less than 21 kg/m(2) were at a significantly higher risk for tibial stress fractures and spinal injuries. Patellofemoral pain syndrome was the most common injury, followed by iliotibial band friction syndrome, plantar fasciitis, meniscal injuries of the knee, and tibial stress syndrome. CONCLUSIONS: Although various risk factors were shown to be positively associated with a risk for, or protection from, specific injuries, future research should include a non-injured control group and a more precise measure of weekly running distance and running experience to validate these results. PMID- 11916891 TI - State anxiety responses to 60 minutes of cross training. AB - OBJECTIVES: Significant reductions in state anxiety following bouts of aerobic exercise have been consistently noted, whereas changes are generally absent after acute resistance training. However, the influence of a single exercise session involving both modes on state anxiety has not been examined. METHODS: To address this, state anxiety responses to 60 minutes of cross training were examined in 16 collegiate athletes (12 women, four men). Each subject completed two cross training exercise sessions (30 minutes of resistance training, 30 minutes of bicycle ergometry) in which the order of the exercises was reversed, with a minimum of one week between sessions. Each exercise mode was completed at about 70% of maximum. State anxiety (SAI-Y1) was assessed five minutes before, and 0, 10, and 60 minutes after exercise. RESULTS: Repeated measures analysis of variance showed a significant (p<0.05) main effect for time. However, the main effect for order and the order by time interaction were not significant. Post hoc analysis showed that state anxiety was reduced (p<0.05) from baseline (mean (SD) = 34.8 (7.9)) at 10 minutes (32.1 (7.5)) and 60 minutes (30.4 (5.9)) after exercise, but not at 0 minutes (33.8 (6.9)). CONCLUSIONS: The results indicate that combined sessions of aerobic and resistance exercise are associated with reductions in state anxiety, and that the order in which the exercise is completed does not influence this response. PMID- 11916892 TI - An ergonomic comparison of rowing machine designs: possible implications for safety. AB - OBJECTIVES: Ergometer training is a common cause of injuries in rowers. A randomised crossover study comparing two power head designs was carried out to examine ergonomic risk factors. METHODS: Six elite male rowers undertook 20 minute fatiguing rowing pieces with both fixed and floating power heads. A CODA MPX infrared telemetric motion analysis detector and the ergometer's interface were used to measure displacement, force, work performed, and power output. RESULTS: There was no significant difference in the total work performed, power per stroke, or metabolic load between the two ergometer designs. Fatigue was shown by a mean (SEM) fall of 9.7 (0.79) W/stroke (95% confidence interval (CI) 8.0 to 11.5) between minutes 8-10 and minutes 16-18 (p<0.001). The stroke length was 53 (13) mm (95% CI 18 to 89) longer with the fixed power head (p<0.02). With fatigue, the stroke with the fixed power head lengthened at the "catch" (beginning of the stroke) by 19.5 mm (p<0.01) and shortened at the finish of the stroke by 7.2 mm (p<0.05). No significant changes in stroke length were seen with the floating power head. The mean force per stroke was 12.1% (95% CI 3.0 to 21.2) (27.3 (8.0) N) higher with the power head fixed versus floating (p<0.02). CONCLUSIONS: It is postulated that longer stroke lengths and greater forces are risk factors for soft tissue injuries. Further research into whether floating power head rowing ergometers are associated with lower injury rates than fixed power head designs is now needed. PMID- 11916893 TI - Lactic acidosis, potassium, and the heart rate deflection point in professional road cyclists. AB - OBJECTIVE: To determine the influence of lactic acidosis, the Bohr effect, and exercise induced hyperkalaemia on the occurrence of the heart rate deflection point (HRDP) in elite (professional) cyclists. METHODS: Sixteen professional male road cyclists (mean (SD) age 26 (1) years) performed a ramp test on a cycle ergometer (workload increases of 5 W/12 s, averaging 25 W/min). Heart rate (HR), gas exchange parameters, and blood variables (lactate, pH, P(50) of the oxyhaemoglobin dissociation curve, and K(+)) were measured during the tests. RESULTS: A HRDP was shown in 56% of subjects at about 88% of their maximal HR (HRDP group; n = 9) but was linear in the rest (No-HRDP group; n = 7). In the HRDP group, the slope of the HR-workload regression line above the HRDP correlated inversely with levels of K(+) at the maximal power output (r = -0.67; p<0.05). CONCLUSIONS: The HRDP phenomenon is associated, at least partly, with exercise induced hyperkalaemia. PMID- 11916894 TI - Management of medical confidentiality in English professional football clubs: some ethical problems and issues. AB - OBJECTIVE: To examine the ways in which confidential matters are dealt with in the context of the relationship between the club doctor (or physiotherapist) and the player as patient in English professional football clubs. METHODS: Semistructured tape recorded interviews with 12 club doctors, 10 club physiotherapists, and 27 current and former players. A questionnaire was also sent to 90 club doctors; 58 were returned. RESULTS: There is among club doctors and physiotherapists no commonly held code of ethics governing how much and what kind of information about players may properly be passed on to managers; associated with this, there is considerable variation from one club to another in terms of the amount and kind of information passed on to managers. In some clubs, medical staff attempt to operate more or less on the basis of the rules governing confidentiality that apply in general practice, but in other clubs, medical staff are more ready to pass on personal information about players. In some situations, this raises serious ethical questions. CONCLUSIONS: Guidelines dealing with confidentiality in practitioner-patient relationships in medical practice have long been available and have recently been restated, specifically in relation to the practice of sports medicine, by the British Olympic Association, the British Medical Association, and the Football Association. This is a welcome first step. However, if the guidelines are to have an impact on practice, detailed consideration needs to be given to ensuring their effective implementation; if this is to be achieved, consideration also needs to be given to identifying those aspects of the culture and organisation of professional football clubs that may hinder the full and effective implementation of those guidelines. PMID- 11916895 TI - The quality of research in sports journals. AB - OBJECTIVE: To examine the evidence base of sports medicine research. METHODS: A sample of four major journals that present core research in sport and exercise medicine (British Journal of Sports Medicine, Medicine and Science in Sports and Exercise, Journal of Sports Medicine and Physical Fitness, and Physical Therapy) was examined using assessment criteria taken from the READER method. RESULTS: Randomised controlled trials comprised 10% or less of all original research articles. Observational/descriptive studies were the most commonly published study design. There was a highly significant difference (p<0.0001) in the contents of the four journals but when they were compared by categorising the better quality methods together (randomised control trial, case-control, and cohort studies), the difference was not significant (p = 0.09). CONCLUSIONS: The overall pattern of publication type seems remarkably stable over medical journals, indicating that the quality of sports medicine research is comparable to that in other specialities. PMID- 11916896 TI - Physical exercise or micronutrient supplementation for the wellbeing of the frail elderly? A randomised controlled trial. AB - OBJECTIVE: To examine the effects of 17 weeks of physical exercise and micronutrient supplementation on the psychological wellbeing of 139 independently living, frail, elderly subjects (inactive, body mass index < or =25 or experiencing weight loss). METHODS: Participants (mean (SD) age 78.5 (5.7)) were randomly assigned to: (a) comprehensive, moderate intensity, group exercise; (b) daily micronutrient enriched foods (25-100% recommended daily amount); (c) both; (d) neither. A social programme and identical regular foods were offered as attention control and placebo. RESULTS: At baseline, moderate to low but significant correlations were found between general wellbeing scores and physical fitness (r = 0.28), functional performance (r = 0.37), and blood concentrations of pyridoxine (r = 0.20), folate (r = 0.25), and vitamin D (r = 0.23) (all p values < or =0.02), but not with physical activity levels and other blood vitamin concentrations. General wellbeing score and self rated health were not responsive to 17 weeks of exercise or nutritional intervention. CONCLUSION: Psychological wellbeing in frail elderly people was not responsive to 17 weeks of intervention with exercise and/or micronutrient enriched foods. The moderate but significant correlations between wellbeing and physical fitness and several blood vitamin concentrations at baseline suggest that changes in wellbeing may occur after long term interventions. PMID- 11916897 TI - Effect of warming up on knee proprioception before sporting activity. AB - BACKGROUND: It is now generally accepted that the ligamentous structures of the knee not only act as mechanical restraints but also have a neurophysiological role in joint function and protection. A report that knee joint laxity increases with exercise raised the question as to whether there is any compensatory change in joint position appreciation. OBJECTIVE: To test whether there is a compensatory mechanism for increased ligamentous laxity during normal levels of activity. METHODS: Joint position appreciation was measured, using a previously reported technique, in the knees of sportsmen at rest and after warm up. RESULTS: Joint position appreciation was found to be significantly more sensitive after warm up (p = 0.005). CONCLUSIONS: These findings indicate that joint position appreciation within the knee accommodates physiological changes within the ligaments and muscles after exercise. PMID- 11916898 TI - Snow sports injuries in Scotland: a case-control study. AB - OBJECTIVES: To examine the incidence and patterns of snow sports injuries at the three largest commercial ski areas in Scotland and to identify factors associated with injury risk. METHODS: A prospective case-control study of all injured people at Cairngorm, Glenshee, and Nevis Range ski areas during the 1999-2000 winter season. Personal details, snow sports related variables, diagnosis, and treatment were recorded. Control data were collected at random from uninjured people at all three areas. Random counts were performed to analyse the composition of the on slope population. RESULTS: A total of 732 injuries were recorded in 674 people. Control data were collected from 336 people. The injury rate for the study was 3.7 injuries per 1000 skier days. Alpine skiers comprised 67% of the on slope population, snowboarders 26%, skiboarders 4%, and telemark skiers 2%. Lower limb injuries and sprains were the commonest injuries in alpine skiers and skiboarders. Snowboarders sustained more injuries to the upper limb and axial areas. Skiboarders and snowboarders had a higher incidence of fractures. After adjustment for other variables, three factors were all independently associated with injury: snowboarding (odds ratio (OR) 4.07, 95% confidence interval (CI) 1.65 to 10.08), alpine skiing (OR 3.82, CI 1.6 to 9.13), and age <16 years (OR 1.9, CI 1.14 to 3.17). More than five days of experience in the current season and at least one week of experience in total had a protective effect against injury. CONCLUSIONS: Despite a change in the composition of the alpine population at Scottish ski areas, the overall rate and pattern of injury are similar to those reported previously in comparable studies. Several factors are associated with an increased risk of injury and should be targeted in future injury prevention campaigns. PMID- 11916900 TI - Acute mountain sickness: an unexpected management problem. PMID- 11916899 TI - Sea level and acute responses to hypoxia: do they predict physiological responses and acute mountain sickness at altitude? AB - OBJECTIVES: To compare a range of physiological responses to acute sea level hypoxia at simulated altitudes with the same physiological responses and acute mountain sickness (AMS) scores measured at altitude (similar to the simulated altitudes) during a 17 day trek in the Himalayas. METHODS: Twenty men and women aged 18-54 years took part in the study. End tidal CO(2) tension (PETCO(2)) and saturated oxygen (SaO(2)) were measured using a capnograph. Observations made at sea level and some simulated altitudes were compared with those in the Himalayas. Pairwise correlations were used to examine the correlation between variables and regression, with 95% prediction intervals providing information on how well one variable could be predicted from another for a given subject. RESULTS: There was only a significant correlation for a few comparisons. The 95% prediction intervals for individual SaO(2) values at a range of simulated altitudes were fairly wide going from +/- 4% to +/- 5%. All of the correlations between laboratory and Himalayan PETCO(2) values were not statistically significant. There was a significant correlation for the relation between SaO(2)and PETCO(2) at sea level for the laboratory data (r=-0.51; p=0.05). For the Himalayan data, there were significant correlations at Namche Bazaar (3450 m; day 3) (r=-0.56; p=0.01) and Dingboche (4300 m; day 6) (r=-0.48; p=0.03). The correlation between SaO(2) and PETCO(2) and AMS was generally poor. CONCLUSIONS: The results indicate that limited information can be gained on a subject's response to altitude by assessing physiological variables at sea level and a range of simulated altitudes before the subject carries out a trek at altitude. PMID- 11916901 TI - Growth hormone and insulin-like growth factor I in a Sydney Olympic gold medallist. AB - An Italian athlete who won a gold medal at the Sydney Olympic Games was studied. She was accused of doping after the finding of high levels of plasma growth hormone (GH) before the Games. She was studied firstly under stressed and then under unstressed conditions. In the first study, GH was measured every 20 minutes for one hour; it was above the normal range in all blood samples, whereas insulin like growth factor I (IGF-I) was normal. In the second study, GH progressively returned to accepted normal levels; IGF-I was again normal. It was concluded that the normal range for GH in athletes must be reconsidered for doping purposes, because athletes are subject to stress and thus to wide variations in GH levels. PMID- 11916902 TI - Exertion induced rhabdomyolysis of the long head of the triceps. AB - The case is reported of bilateral rhabdomyolysis of the long head of the triceps following intensive exercise in a 30 year old male weightlifter. The diagnosis was based on myalgias localised to one muscle and raised levels of muscle enzymes. Magnetic resonance imaging helped to locate the site and extent of muscular involvement. Treatment consisted of complete rest and adequate intravenous perfusion to allow clearance of the clinical and biological abnormalities and prevent renal involvement. PMID- 11916903 TI - Simultaneous bilateral quadriceps tendon rupture while playing basketball. AB - Simultaneous bilateral quadriceps tendon rupture is an uncommon injury in healthy people and only a few cases have been reported in athletes. This is the first report of a patient with simultaneous bilateral quadriceps tendon rupture incurred while playing basketball. The injury was surgically repaired and the patient had a good functional outcome. PMID- 11916904 TI - Fructose amplifies counterregulatory responses to hypoglycemia in humans. AB - Glucokinase (GK) is required for cellular glucose sensing, although there is a paucity of data regarding its role in the counterregulatory response to hypoglycemia in humans. Because fructose has been shown to modulate GK activity, we examined the effects of an acute infusion of fructose on hypoglycemia counterregulation in seven lean nondiabetic subjects. Using stepped hypoglycemia clamp studies (5.0, 4.4, 3.9, and 3.3 mmol/l target plasma glucose steps, 50 min each), subjects were studied on two separate occasions, without (control) or with co-infusion of fructose (1.2 mg.kg(-1).min(-1)). Fructose induced a resetting of the glycemic thresholds for secretion of epinephrine (3.8 +/- 0.1 mmol/l) and glucagon (3.9 +/- 0.2 mmol/l) to higher plasma glucose concentrations (4.0 +/- 0.1 mmol/l [P = 0.006] and 4.1 +/- 0.1 mmol/l [P = 0.03], respectively). In addition, the magnitude of increase in epinephrine and glucagon concentrations was higher after administration of fructose (48 and 39%, respectively, P < 0.05 for both). The amplification of these hormonal responses was specific because plasma norepinephrine, growth hormone, and cortisol were comparable in both sets of studies. Endogenous glucose production, measured with [3-(3)H]glucose, increased by 47% (P < 0.05) in the fructose infusion studies compared with 14% (P = NS) in the control studies. In addition, glucose uptake was more suppressed with fructose infusion (by 33%, P < 0.05). In concert with these effects of fructose on glucose kinetics, average glucose infusion rate was markedly reduced in the fructose infusion studies during the 3.9-mmol/l glucose step (4.6 +/- 0.9 vs. 7.4 +/- 1.1 micromol.kg(-1).min(-1), respectively, P = 0.03) and during the 3.3-mmol/l glucose step (0.5 +/- 0.1 vs. 5.2 +/- 1.2 micromol.kg(-1).min(-1), respectively, P < 0.001), suggesting more potent glucose counterregulation and improved recovery from hypoglycemia with fructose infusion. We conclude that infusion of a catalytic dose of fructose amplifies the counterregulatory response to hypoglycemia by both increases in hormonal activation and augmentation of glucose counterregulation in humans. PMID- 11916905 TI - Peroxisome proliferator-activated receptor-alpha regulates fatty acid utilization in primary human skeletal muscle cells. AB - In humans, skeletal muscle is a major site of peroxisome proliferator-activated receptor-alpha (PPAR-alpha) expression, but its function in this tissue is unclear. We investigated the role of hPPAR-alpha in regulating muscle lipid utilization by studying the effects of a highly selective PPAR-alpha agonist, GW7647, on [(14)C]oleate metabolism and gene expression in primary human skeletal muscle cells. Robust induction of PPAR-alpha protein expression occurred during muscle cell differentiation and corresponded with differentiation-dependent increases in oleate oxidation. In mature myotubes, 48-h treatment with 10-1,000 nmol/l GW7647 increased oleate oxidation dose-dependently, up to threefold. Additionally, GW7647 decreased oleate esterification into myotube triacylglycerol (TAG), up to 45%. This effect was not abolished by etomoxir, a potent inhibitor of beta-oxidation, indicating that PPAR-alpha-mediated TAG depletion does not depend on reciprocal changes in fatty acid catabolism. Consistent with its metabolic actions, GW7647 induced mRNA expression of mitochondrial enzymes that promote fatty acid catabolism; carnitine palmityltransferase 1 and malonyl-CoA decarboxylase increased approximately 2-fold, whereas pyruvate dehydrogenase kinase 4 increased 45-fold. Expression of several genes that regulate glycerolipid synthesis was not changed by GW7647 treatment, implicating involvement of other targets to explain the TAG-depleting effect of the compound. These results demonstrate a role for hPPAR-alpha in regulating muscle lipid homeostasis. PMID- 11916906 TI - Thyroid hormone receptor interacting protein 3 (trip3) is a novel coactivator of hepatocyte nuclear factor-4alpha. AB - Mutations of the hepatocyte nuclear factor-4alpha (HNF-4alpha) gene are associated with a subtype of maturity-onset diabetes of the young (MODY1) that is characterized by impaired insulin secretion in response to a glucose load. HNF 4alpha, which is a transcription factor expressed in pancreatic beta-cells, plays an important role in regulating the expression of genes involved in glucose metabolism. Thus, cofactors that interact with HNF-4alpha and modify its transcriptional activity might also play an important role in regulating the metabolic pathways in pancreatic beta-cells, and the genes of such cofactors are plausible candidate genes for MODY. In the present study, we showed, using a yeast two-hybrid screening assay, that thyroid hormone receptor interacting protein 3 (Trip3) interacted with HNF-4alpha, and their interaction was confirmed by the glutathione S-transferase pull-down assay. Human Trip3 cDNA contained an open reading frame for a protein of 155 amino acids, and the gene was expressed in both pancreatic islets and MIN6 cells. Cotransfection experiments indicated that Trip3 could enhance (two- to threefold) the transcription activity of HNF 4alpha in COS-7 cells and MIN6 cells. These results suggest that Trip3 is a coactivator of HNF-4alpha. Mutation screening revealed that variation of the Trip3 gene is not a common cause of MODY/early-onset type 2 diabetes in Japanese individuals. Trip3 may play an important role in glucose metabolism by regulating the transcription activity of HNF-4alpha. PMID- 11916907 TI - Independent regulation of in vivo insulin action on glucose versus K(+) uptake by dietary fat and K(+) content. AB - Insulin stimulates both glucose and K(+) uptake, and high-fat feeding is known to decrease insulin-stimulated glucose uptake. The purpose of this study was to examine whether insulin's actions on glucose and K(+) uptake are similarly decreased by a high-fat diet. Wistar rats were fed a standard control (12.2% fat; n = 6) or high-fat (66.5% fat; n = 13) diet for 15 days. Because K(+) content was 1% in the control and 0.5% in the high-fat diet and because the rats ate less of the high-fat diet, we also compared the high-fat diet with 0.5% K(+) (HFD; n = 7) to a high-fat diet supplemented with 1.5% K(+) (HFD+K; n = 6). K(+) intake was matched between the control and HFD+K groups (246 +/- 8 vs. 224 +/- 2 mg/day), but was lower in the HFD group (78 +/- 10 mg/day; P < 0.05). Insulin-stimulated glucose and K(+) uptake were determined by hyperinsulinemic (5 mU.kg(-1).min(-1)) glucose and K(+) clamps. The HFD depressed both insulin-stimulated glucose uptake compared to the control (133 +/- 5 vs. 166 +/- 7 micromol.kg(-1).min(-1); P < 0.05) and K(+) uptake (5.5 +/- 0.9 vs. 8.9 +/- 1.0 micromol.kg(-1).min(-1); P < 0.05) compared to the control. However, insulin-stimulated K(+) uptake was unchanged in the HFD+K versus in the control group (10.0 +/- 0.6 vs. 8.9 +/- 1.0 micromol.kg(-1).min(-1); P > 0.05), whereas insulin-stimulated glucose uptake in the HFD+K group was decreased to a rate (137 +/- 9 micromol.kg(-1).min(-1)), similar to that of the HFD group. We concluded that the decrease in insulin stimulated K(+) uptake during high-fat feeding was a result of decreased K(+) intake, and that insulin's actions on glucose uptake and K(+) uptake are independently regulated by dietary fat and K(+) content, respectively. PMID- 11916908 TI - The diabetic phenotype is conserved in myotubes established from diabetic subjects: evidence for primary defects in glucose transport and glycogen synthase activity. AB - The most well-described defect in the pathophysiology of type 2 diabetes is reduced insulin-mediated glycogen synthesis in skeletal muscles. It is unclear whether this defect is primary or acquired secondary to dyslipidemia, hyperinsulinemia, or hyperglycemia. We determined the glycogen synthase (GS) activity; the content of glucose-6-phosphate, glucose, and glycogen; and the glucose transport in satellite cell cultures established from diabetic and control subjects. Myotubes were precultured in increasing insulin concentrations for 4 days and subsequently stimulated acutely by insulin. The present study shows that the basal glucose uptake as well as insulin-stimulated GS activity is reduced in satellite cell cultures established from patients with type 2 diabetes. Moreover, increasing insulin concentrations could compensate for the reduced GS activity to a certain extent, whereas chronic supraphysiological insulin concentrations induced insulin resistance in GS and glucose transport activity. Our data suggest that insulin resistance in patients with type 2 diabetes comprises at least two important defects under physiological insulin concentrations: a reduced glucose transport under basal conditions and a reduced GS activity under acute insulin stimulation, implicating a reduced glucose uptake in the fasting state and a diminished insulin-mediated storage of glucose as glycogen after a meal. PMID- 11916909 TI - Orally administered leucine enhances protein synthesis in skeletal muscle of diabetic rats in the absence of increases in 4E-BP1 or S6K1 phosphorylation. AB - In this study, food-deprived (18 h) control rats and rats with alloxan-induced diabetes were orally administered saline or the amino acid leucine to assess whether it regulates protein synthesis independently of a change in serum insulin concentrations. Immediately after leucine administration, diabetic rats were infused with insulin (0.0, 4.0, or 20 pmol small middle dot min(-1) small middle dot kg(-1)) for 1 h to examine the role of the hormone in the protein synthetic response to leucine. In control rats, leucine stimulated protein synthesis by 58% and increased phosphorylation of the translational repressor, eukaryotic initiation factor (eIF) 4E-binding protein (BP)-1, 4E-BP1, fivefold. Consequently, association of the mRNA cap-binding protein eukaryotic initiation factor (eIF)4E with 4E-BP1 was reduced to 50% of control values, and eIF4G*eIF4E complex assembly was increased 80%. Furthermore, leucine increased the phosphorylation of the 70-kDa ribosomal protein S6 (rp S6) and the ribosomal protein S6 kinase (S6K1). Diabetes attenuated protein synthesis compared with control rats. Nonetheless, in diabetic rats, leucine increased protein synthesis by 53% without concomitant changes in the phosphorylation of 4E-BP1 or S6K1. Skeletal muscle protein synthesis was stimulated in diabetic rats infused with insulin, but rates of synthesis remained less than values in nondiabetic controls that were administered leucine. Phosphorylation of 4E-BP1 and S6K1 was increased in diabetic rats infused with insulin in a dose-dependent manner, and the response was enhanced by leucine. The results suggest that leucine enhances protein synthesis in skeletal muscle through both insulin-dependent and independent mechanisms. The insulin-dependent mechanism is associated with increased phosphorylation of 4E-BP1 and S6K1. In contrast, the insulin independent effect on protein synthesis is mediated by an unknown mechanism. PMID- 11916910 TI - Indinavir induces acute and reversible peripheral insulin resistance in rats. AB - The use of HIV protease inhibitors (PIs) has been associated with several metabolic changes, including lipodystrophy, hyperlipidemia, and insulin resistance. The etiology of these adverse effects remains unknown. PIs have recently been found to cause acute and reversible inhibition of GLUT4 activity in vitro. To determine the acute in vivo effects of indinavir on whole-body glucose homeostasis, glucose tolerance tests were performed on PI-naive Wistar rats immediately after a single intravenous dose of indinavir. Glucose and insulin levels were significantly elevated in indinavir-treated versus control rats (P < 0.05) during the initial 30 min of the glucose tolerance test. Under euglycemic- hyperinsulinemic clamp conditions, indinavir treatment acutely reduced the glucose infusion rate required to maintain euglycemia by 18 and 49% at indinavir concentrations of 14 and 27 micromol/l, respectively. Muscle 2-deoxyglucose uptake was similarly reduced under these conditions. Restoration of insulin sensitivity was observed within 4 h after stopping the indinavir infusion. Indinavir did not alter the suppression of hepatic glucose output under hyperinsulinemic conditions. These data demonstrate that indinavir causes acute and reversible changes in whole-body glucose homeostasis in rats and support the contribution of GLUT4 inhibition to the development of insulin resistance in patients treated with PIs. PMID- 11916911 TI - Long-term treatment with the dipeptidyl peptidase IV inhibitor P32/98 causes sustained improvements in glucose tolerance, insulin sensitivity, hyperinsulinemia, and beta-cell glucose responsiveness in VDF (fa/fa) Zucker rats. AB - The incretins, glucose-dependent insulinotropic polypeptide (GIP) and glucagon like peptide 1 (GLP-1) are responsible for >50% of nutrient-stimulated insulin secretion. After being released into the circulation, GIP and GLP-1 are rapidly inactivated by the circulating enzyme dipeptidyl peptidase IV (DP IV). The use of DP IV inhibitors to enhance these insulinotropic hormonal axes has proven effective on an acute scale in both animals and humans; however, the long-term effects of these compounds have yet to be determined. Therefore, we carried out the following study: two groups of fa/fa Zucker rats (n = 6 each) were treated twice daily for 3 months with the DP IV inhibitor P32/98 (20 mg.kg(-1).day(-1), p.o.). Monthly oral glucose tolerance tests (OGTTs), performed after drug washout, revealed a progressive and sustained improvement in glucose tolerance in the treated animals. After 12 weeks of treatment, peak OGTT blood glucose values in the treated animals averaged 8.5 mmol/l less than in the controls (12.0 +/- 0.7 vs. 20.5 +/- 1.3 mmol/l, respectively). Concomitant insulin determinations showed an increased early-phase insulin response in the treated group (43% increase). Furthermore, in response to an 8.8 mmol/l glucose perfusion, pancreata from controls showed no increase in insulin secretion, whereas pancreata from treated animals exhibited a 3.2-fold rise in insulin secretion, indicating enhanced beta-cell glucose responsiveness. Also, both basal and insulin stimulated glucose uptake were increased in soleus muscle strips from the treated group (by 20 and 50%, respectively), providing direct evidence for an improvement in peripheral insulin sensitivity. In summary, long-term DP IV inhibitor treatment was shown to cause sustained improvements in glucose tolerance, insulinemia, beta-cell glucose responsiveness, and peripheral insulin sensitivity, novel effects that provide further support for the use of DP IV inhibitors in the treatment of diabetes. PMID- 11916912 TI - Impaired beta-cell function, incretin effect, and glucagon suppression in patients with type 1 diabetes who have normal fasting glucose. AB - We have recently described a novel phenotype in a group of subjects with type 1 diabetes that is manifested by glucose >11.1 mmol/l 120 min after an oral glucose load, but with normal fasting glucose levels. We now describe the metabolic characteristics of these subjects by comparing parameters of islet hormone secretion and glucose disposal in these subjects to age-matched nondiabetic control subjects. The patients with type 1 diabetes had fasting glucose, insulin, and glucagon values similar to those of control subjects. Additionally, the insulin secretory response to intravenous arginine at euglycemia was similar in the control and diabetic groups (264 +/- 33.5 and 193 +/- 61.3 pmol/l; P = 0.3). However, marked differences in beta-cell function were found in response to hyperglycemia. Specifically, the first-phase insulin response was lower in diabetic subjects (329.1 +/- 39.6 vs. 91.3 +/- 34.1 pmol/l; P < 0.001), as was the slope of glucose potentiation of the insulin response to arginine (102 +/- 18.7 vs. 30.2 +/- 6.1 pmol/l per mmol/l; P = 0.005) and the maximum insulin response to arginine (2,524 +/- 413 vs. 629 +/- 159 pmol/l; P = 0.001). Although plasma levels of glucagon-like peptide (GLP)-1 and gastric inhibitory peptide (GIP) did not differ between control and diabetic subjects, the incretin effect was lower in the diabetic patients (70.3 +/- 5.4 vs. 52.1 +/- 5.9%; P = 0.03). Finally, there was a lack of suppression of glucagon in the patients after both oral and intravenous glucose administration, which may have contributed to their postprandial hyperglycemia. Glucose effectiveness did not differ between patients and control subjects, nor did insulin sensitivity, although there was a tendency for the patients to be insulin resistant (9.18 +/- 1.59 vs. 5.22 +/- 1.17 pmol.( 1).min(-1); P = 0.08). These data characterize a novel group of subjects with type 1 diabetes manifested solely by hyperglycemia following an oral glucose load in whom islet function is normal at euglycemia, but who have marked defects in both alpha- and beta-cell secretion at hyperglycemia. This pattern of abnormalities may be characteristic of islet dysfunction early in the development of type 1 diabetes. PMID- 11916913 TI - Intraislet hyperinsulinemia prevents the glucagon response to hypoglycemia despite an intact autonomic response. AB - Because absence of the glucagon response to falling plasma glucose concentrations plays a key role in the pathogenesis of iatrogenic hypoglycemia in patients with insulin-deficient diabetes and the mechanism of this defect is unknown, and given evidence in experimental animals that a decrease in intraislet insulin is a signal to increased glucagon secretion, we examined the role of endogenous insulin in the physiological glucagon response to hypoglycemia. We tested the hypothesis that intraislet hyperinsulinemia prevents the glucagon response to hypoglycemia despite an intact autonomic-adrenomedullary, sympathetic neural, and parasympathetic neural-response and a low alpha-cell glucose concentration. Twelve healthy young adults were studied on three separate occasions. Insulin was infused in hourly steps in relatively low doses (1.5, 3.0, 4.5, and 6.0 pmol.kg( 1).min(-1)) from 60 through 300 min on all three occasions. Plasma glucose levels were clamped at euglycemia ( approximately 5.0 mmol/l, approximately 90 mg/dl) on one occasion and at hourly steps of approximately 4.7, 4.2, 3.6, and 3.0 mmol/l ( approximately 85, 75, 65, and 55 mg/dl) from 60 through 300 min on the other two occasions. On one of the latter occasions, the beta-cell secretagogue tolbutamide was infused in a dose of 1.0 g/h from 60 through 300 min. Hypoglycemia with tolbutamide infusion, compared with similar hypoglycemia alone, was associated with higher (P < 0.0001) C-peptide levels (final values of 1.0 +/- 0.2 vs. 0.1 +/ 0.0 nmol/l), higher (P < 0.0001) rates of insulin secretion (final values of 198 +/- 60 vs. 15 +/- 4 pmol/min), and higher (P < 0.0001) insulin levels (final values of 325 +/- 30 vs. 245 +/- 20 pmol/l) as expected. The glucagon response to hypoglycemia was prevented during tolbutamide infusion (P < 0.0001). Glucagon levels were 17 +/- 1 pmol/l at baseline on both occasions, 14 +/- 1 vs. 15 +/- 1 pmol/l, respectively, during the initial hyperinsulinemic euglycemia, and 15 +/- 1 vs. 22 +/- 2 pmol/l, respectively, during hypoglycemia with and without tolbutamide infusion. Autonomic-adrenomedullary (plasma epinephrine), sympathetic neural (plasma norepinephrine), and parasympathetic neural (plasma pancreatic polypeptide)-responses to hypoglycemia were not reduced during tolbutamide infusion. We conclude that intraislet hyperinsulinemia prevents the glucagon response to hypoglycemia despite an intact autonomic response and a low alpha cell glucose concentration. PMID- 11916914 TI - Activation of IRS-2-mediated signal transduction by IGF-1, but not TGF-alpha or EGF, augments pancreatic beta-cell proliferation. AB - Transforming growth factor (TGF)-alpha- and epidermal growth factor (EGF)-induced signal transduction was directly compared with that of glucose and insulin-like growth factor-1 (IGF-1) in INS-1 cells. TGF-alpha/EGF transiently (<20 min) induced phosphorylation of extracellular-regulated kinase (Erk)-1/2 (>20-fold), glycogen synthase kinase (GSK)-3 (>10-fold), and protein kinase B (PKB) (Ser(473) and Thr(308)), but did not increase [(3)H]thymidine incorporation. In contrast, phosphorylation of Erk1/2, GSK-3, and PKB in response to glucose and IGF-1 was more prolonged (>24 h) and, though not as robust as TGF-alpha/EGF, did increase beta-cell proliferation. Phosphorylation of p70(S6K) was also increased by IGF 1/glucose, but not by TGF-alpha/EGF, despite upstream PKB activation. It was found that IGF-1 induced phosphatidylinositol 3-kinase (PI3K) association with insulin receptor substrate (IRS)-1 and -2 in a glucose-dependent manner, whereas TGF-alpha/EGF did not. The importance of specific IRS-2-mediated signaling events was emphasized in that adenoviral-mediated overexpression of IRS-2 further increased glucose/IGF-1-induced beta-cell proliferation (more than twofold; P < 0.05) compared with control or adenoviral-mediated IRS-1 overexpressing INS-1 cells. Neither IRS-1 nor IRS-2 overexpression induced a beta-cell proliferative response to TGF-alpha/EGF. Thus, a prolonged activation of Erk1/2 and PI3K signaling pathways is important in committing a beta-cell to a mitogenic event, and it is likely that this sustained activation is instigated by signal transduction occurring specifically through IRS-2. PMID- 11916915 TI - Expression profiling of palmitate- and oleate-regulated genes provides novel insights into the effects of chronic lipid exposure on pancreatic beta-cell function. AB - Chronic lipid exposure is implicated in beta-cell dysfunction in type 2 diabetes. We therefore used oligonucleotide arrays to define global alterations in gene expression in MIN6 cells after 48-h pretreatment with oleate or palmitate. Altogether, 126 genes were altered > or =1.9-fold by palmitate, 62 by oleate, and 46 by both lipids. Importantly, nine of the palmitate-regulated genes are known to be correspondingly changed in models of type 2 diabetes. A tendency toward beta-cell de-differentiation was also apparent with palmitate: pyruvate carboxylase and mitochondrial glycerol 3-phosphate dehydrogenase were downregulated, whereas lactate dehydrogenase and fructose 1,6-bisphosphatases were induced. Increases in the latter (also seen with oleate), along with glucosamine-phosphate N-acetyl transferase, imply upregulation of the hexosamine biosynthesis pathway in palmitate-treated cells. However, palmitate also increased expression of calcyclin and 25-kDa synaptosomal-associated protein (SNAP25), which control distal secretory processes. Consistent with these findings, secretory responses to noncarbohydrate stimuli, especially palmitate itself, were upregulated in palmitate-treated cells (much less so with oleate). Indeed, glucose-stimulated secretion was slightly sensitized by chronic palmitate exposure but inhibited by oleate treatment, whereas both lipids enhanced basal secretion. Oleate and palmitate also induced expression of chemokines (MCP-1 and GRO1 oncogene) and genes of the acute phase response (serum amyloid A3). Increases in transcriptional modulators such as ATF3, CCAAT/enhancer binding protein-beta (C/EBPbeta), C/EBPdelta, and c-fos were also seen. The results highlight links between regulated gene expression and phenotypic alterations in palmitate versus oleate-pretreated beta-cells. PMID- 11916916 TI - Pulsatile insulin release from islets isolated from three subjects with type 2 diabetes. AB - Plasma insulin in healthy subjects shows regular oscillations, which are important for the hypoglycemic action of the hormone. In individuals with type 2 diabetes, these regular variations are altered, which has been implicated in the development of insulin resistance and hyperglycemia. The origin of the change is unknown, but derangement of the islet secretory pattern has been suggested as a contributing cause. In the present study, we show the dynamics of insulin release from individually perifused islets isolated from three subjects with type 2 diabetes. Insulin release at 3 mmol/l glucose was 10.5 +/- 4.5 pmol.g(-1).s(-1) and pulsatile (0.26 +/- 0.05 min(-1)). In islets from one subject, 11 mmol/l glucose transiently increased insulin release by augmentation of the insulin pulses without affecting the frequency. Addition of 1 mmol/l tolbutamide did not increase insulin release. In islets from the remaining subjects, insulin release was not affected by 11 mmol/l glucose. Tolbutamide transiently increased insulin release in islets from one subject. Insulin release from four normal subjects at 3 mmol/l glucose was 4.3 +/- 0.8 pmol.g(-1).s(-1) and pulsatile (0.23 +/- 0.03 min(-1)). At 11 mmol/l glucose, insulin release increased in islets from all subjects. Tolbutamide further increased insulin release in islets from two subjects. It is concluded that islets from the three individuals with type 2 diabetes release insulin in pulses. The impaired secretory response to glucose may be related to impaired metabolism before mitochondrial degradation of the sugar. PMID- 11916917 TI - Carbon monoxide protects pancreatic beta-cells from apoptosis and improves islet function/survival after transplantation. AB - Pancreatic islets transplanted to treat autoimmune type 1 diabetes often fail to function (primary nonfunction), likely because of islet beta-cell apoptosis. We show that carbon monoxide (CO), a product of heme oxygenase activity, protects beta-cells from apoptosis. Protection is mediated through guanylate cyclase activation, generation of cyclic GMP (cGMP), and activation of cGMP-dependent protein kinases. This antiapoptotic effect is still observed when beta-cells are exposed to CO for 1 h before the apoptotic stimulus. In a similar manner, mouse islets exposed to CO for just 2 h function significantly better after transplantation than islets not exposed to CO. These findings suggest a potential therapeutic application for CO in improving islet function/survival after transplantation in humans. PMID- 11916918 TI - Blood letting in high-ferritin type 2 diabetes: effects on insulin sensitivity and beta-cell function. AB - Iron-related insulin-resistance is improved by iron depletion or treatment with iron chelators. The aim of this study was to evaluate insulin sensitivity and insulin secretion after blood letting in patients who had high-ferritin type 2 diabetes and were randomized to blood letting (three phlebotomies [500 ml of blood] at 2-week intervals, group 1) or to observation (group 2). Insulin secretion and sensitivity were tested at baseline and 4 and 12 months thereafter. The two groups were matched for age, BMI, pharmacologic treatment, and chronic diabetic complications. All patients were negative for C282Y mutation of hereditary hemochromatosis. Baseline glycated hemoglobin (6.27 +/- 0.9% vs. 6.39 +/- 1.2%), insulin sensitivity (2.75 +/- 1.8 vs. 3.2 +/- 2.1 mg.dl(-1).min(-1)), and area under the curve for C-peptide (AUC(C-peptide); 38.7 +/- 11.6 vs. 37.6 +/ 14.1 ng.ml(-1).min(-1)) were not significantly different between the two groups of patients. Body weight, blood pressure, blood hematocrit levels, and drug treatment remained essentially unchanged during the study period. As expected, serum ferritin, transferrin saturation index, and blood hemoglobin decreased significantly at 4 months only in patients who received blood letting. In parallel to this changes, blood HbA(1c) decreased significantly only in group 1 subjects (mean differences, -0.61; 95% CI, -0.17 to -1.048; P = 0.01). AUC(C peptide) decreased by -10.2 +/- 6.3% after blood letting. In contrast, a 10.4 +/- 6.4% increase in AUC(C-peptide) was noted in group 2 subjects at 4 months (P = 0.032). At 12 months, AUC(C-peptide) returned to values not significantly different from baseline in the two groups of subjects. At 4 months, the change in insulin sensitivity from baseline was significantly different between the two groups (80.6 +/- 43.2% vs. -8.6 +/- 9.9% in groups 1 and 2, respectively, P = 0.049). At 12 months, the differences between the two groups were even more marked (55.5 +/- 24.8% vs. -26.8 +/- 9.9%; P = 0.005). When the analysis was restricted to those subjects who completed the follow-up until 12 months, results did not show differences compared with the changes observed at 4 months, except for insulin sensitivity. A statistically significant increase in insulin sensitivity was observed in the blood-letting group (from 2.30 +/- 1.81 to 3.08 +/- 2.55 mg.dl(-1).min(-1) at 4 months, to 3.16 +/- 1.85 mg.dl(-1).min(-1) at 12 months; P = 0,045) in contrast with group 2 subjects (from 3.24 +/- 1.9 to 3.26 +/- 2.05 mg.dl(-1).min(-1) at 4 months, to 2.31 +/- 1.35 mg.dl(-1).min(-1) at 12 months). In summary, blood letting led simultaneously to decreased blood HbA(1c) levels and to changes in insulin secretion and insulin resistance that were significantly different from those observed in a matched observational group of subjects with high-ferritin type 2 diabetes. The mechanisms for improvement in peripheral insulin sensitivity after blood letting should be investigated further. PMID- 11916919 TI - The concurrent accumulation of intra-abdominal and subcutaneous fat explains the association between insulin resistance and plasma leptin concentrations : distinct metabolic effects of two fat compartments. AB - Obesity is associated with insulin resistance, particularly when body fat has a central distribution. However, insulin resistance also frequently occurs in apparently lean individuals. It has been proposed that these lean insulin resistant individuals have greater amounts of body fat than lean insulin sensitive subjects. Alternatively, their body fat distribution may be different. Obesity is associated with elevated plasma leptin levels, but some studies have suggested that insulin sensitivity is an additional determinant of circulating leptin concentrations. To examine how body fat distribution contributes to insulin sensitivity and how these variables are related to leptin levels, we studied 174 individuals (73 men, 101 women), a priori classified as lean insulin sensitive (LIS, n = 56), lean insulin-resistant (LIR, n = 61), and obese insulin resistant (OIR, n = 57) based on their BMI and insulin sensitivity index (S(I)). Whereas the BMI of the two lean groups did not differ, the S(I) of the LIR subjects was less than half that of the LIS group. The subcutaneous and intra abdominal fat areas, determined by computed tomography, were 45 and 70% greater in the LIR subjects (P < 0.001) and 2.5- and 3-fold greater in the OIR group, as compared with the LIS group. Fasting plasma leptin levels were moderately increased in LIR subjects (10.8 +/- 7.1 vs. 8.1 +/- 6.4 ng/ml in LIS subjects; P < 0.001) and doubled in OIR subjects (21.9 +/- 15.5 ng/ml; P < 0.001). Because of the confounding effect of body fat, we examined the relationships between adiposity, insulin sensitivity, and leptin concentrations by multiple regression analysis. Intra-abdominal fat was the best variable predicting insulin sensitivity in both genders and explained 54% of the variance in S(I). This inverse relationship was nonlinear (r = -0.688). On the other hand, in both genders, fasting leptin levels were strongly associated with subcutaneous fat area (r = 0.760) but not with intra-abdominal fat. In line with these analyses, when LIS and LIR subjects were matched for subcutaneous fat area, age, and gender, they had similar leptin levels, whereas their intra-abdominal fat and insulin sensitivity remained different. Thus, accumulation of intra-abdominal fat correlates with insulin resistance, whereas subcutaneous fat deposition correlates with circulating leptin levels. We conclude that the concurrent increase in these two metabolically distinct fat compartments is a major explanation for the association between insulin resistance and elevated circulating leptin concentrations in lean and obese subjects. PMID- 11916920 TI - Leptin resistance during aging is independent of fat mass. AB - Increased fat mass, abdominal adiposity, and insulin resistance are typical findings in aging mammals and are frequently associated with leptin resistance and increased plasma leptin levels. To examine whether leptin's failure in aging is due to aging per se or to changes in body fat mass or distribution, we studied aging rats that underwent calorie restriction throughout their lives, maintaining their youthful body fat pattern and metabolic profile. Leptin's action was assessed by measuring its ability to regulate food intake, fat mass and its distribution, peripheral and hepatic insulin action, and its own gene expression in fat. Our results show that leptin's action is markedly diminished in aging rats, independently of their body fat pattern. Leptin's failure in this model suggests its causative role in the metabolic decline seen with aging. PMID- 11916921 TI - Assessment of skeletal muscle triglyceride content by (1)H nuclear magnetic resonance spectroscopy in lean and obese adolescents: relationships to insulin sensitivity, total body fat, and central adiposity. AB - The metabolism and composition of skeletal muscle tissue is of special interest because it is a primary site of insulin action and plays a key role in the pathogenesis of insulin resistance. Intramyocellular (IMCL) triglyceride stores are an accessible form of energy that may decrease skeletal muscle glucose utilization, thereby contributing to impaired glucose metabolism. Because of the invasive nature of muscle biopsies, there is limited, if any, information about intramuscular lipid stores in children. The development of (1)H nuclear magnetic resonance (NMR) spectroscopy provides a unique noninvasive alternative method that differentiates intracellular fat from intercellular fat in muscle tissue. The present study was performed to determine whether IMCL and extramyocellular (EMCL) lipid contents are increased early in the development of juvenile obesity and to explore the relationships between IMCL and EMCL to in vivo insulin sensitivity, independently of total body fat and central adiposity in obese and nonobese adolescents. Eight nonobese (BMI 21 kg/m(2), age 11-16 years) and 14 obese (BMI 35 +/- 1.5 kg/m(2), age 11-15 years) adolescents underwent 1) (1)H-NMR spectroscopy to noninvasively quantify IMCL and EMCL triglyceride content of the soleus muscle, 2) a 2-h euglycemic-hyperinsulinemic clamp (40 mU.m(-2).min(-1)) to assess insulin sensitivity, 3) a dual-energy X-ray absorptiometry scan to measure total percent body fat, and 4) magnetic resonance imaging to measure abdominal fat distribution. Both the IMCL and EMCL content of the soleus muscle were significantly greater in the obese adolescents than in the lean control subjects. A strong inverse correlation was found between IMCL and insulin sensitivity, which persisted and became even stronger after controlling for percent total body fat and abdominal subcutaneous fat mass (partial correlation r = -0.73, P < 0.01) but not when adjusting for visceral fat (r = - 0.54, P < 0.08). In obese adolescents, increase in total body fat and central adiposity were accompanied by higher IMCL and EMCL lipid stores. The striking relationships between both IMCL and EMCL with insulin sensitivity in childhood suggest that these findings are not a consequence of aging but occur early in the natural course of obesity. PMID- 11916922 TI - Specific inhibition of PTEN expression reverses hyperglycemia in diabetic mice. AB - Signaling through the phosphatidylinositol 3'-kinase (PI3K) pathway is crucial for metabolic responses to insulin, and defects in PI3K signaling have been demonstrated in type 2 diabetes. PTEN (MMAC1) is a lipid/protein phosphatase that can negatively regulate the PI3K pathway by dephosphorylating phosphatidylinositol (3,4,5)-triphosphate, but it is unclear whether PTEN is physiologically relevant to insulin signaling in vivo. We employed an antisense oligonucleotide (ASO) strategy in an effort to specifically inhibit the expression of PTEN. Transfection of cells in culture with ASO targeting PTEN reduced PTEN mRNA and protein levels and increased insulin-stimulated Akt phosphorylation in alpha-mouse liver-12 (AML12) cells. Systemic administration of PTEN ASO once a week in mice suppressed PTEN mRNA and protein expression in liver and fat by up to 90 and 75%, respectively, and normalized blood glucose concentrations in db/db and ob/ob mice. Inhibition of PTEN expression also dramatically reduced insulin concentrations in ob/ob mice, improved the performance of db/db mice during insulin tolerance tests, and increased Akt phosphorylation in liver in response to insulin. These results suggest that PTEN plays a significant role in regulating glucose metabolism in vivo by negatively regulating insulin signaling. PMID- 11916923 TI - Human obesity and type 2 diabetes are associated with alterations in SREBP1 isoform expression that are reproduced ex vivo by tumor necrosis factor-alpha. AB - Sterol regulatory element binding protein (SREBP)-1 is a transcription factor with important roles in the control of fatty acid metabolism and adipogenesis. Little information is available regarding the expression of this molecule in human health or disease. Exposure of isolated human adipocytes to insulin enhanced SREBP1 gene expression and promoted its proteolytic cleavage to the active form. Furthermore, 3 h of in vivo hyperinsulinemia also significantly increased SREBP1 gene expression in human skeletal muscle. Transcript levels of SREBP1c, the most abundant isoform in adipose tissue, were significantly decreased in the subcutaneous adipose tissue of obese normoglycemic and type 2 diabetic subjects compared with that of nonobese normoglycemic control subjects. In skeletal muscle, SREBP1 expression was significantly reduced in type 2 diabetic subjects but not in obese subjects. Within the diabetic group, the extent of SREBP1 suppression was inversely related to metabolic control and was normalized by 3 h of in vivo hyperinsulinemia. Exposure of isolated human adipocytes to tumor necrosis factor-alpha (TNF-alpha) produced a marked and specific decrease in the mRNA encoding the SREBP1c isoform and completely blocked the insulin-induced cleavage of SREBP1 protein. Thus, both the expression and proteolytic maturation of human SREBP1 are positively modulated by insulin. The specific reduction in the SREBP1c isoform seen in the adipose tissue of obese and type 2 diabetic subjects can be recapitulated ex vivo by TNF-alpha, suggesting a possible mechanism for this association. PMID- 11916924 TI - Novel genes regulated by the insulin sensitizer rosiglitazone during adipocyte differentiation. AB - Thiazolidinediones (TZDs) are a new class of compounds that improve insulin sensitivity in type 2 diabetic patients as well as in rodent models of this disease. These compounds act as ligands for a member of the nuclear hormone receptor superfamily, peroxisome proliferator-activated receptor-gamma (PPAR gamma), which is highly expressed in adipose tissue and, moreover, has been shown to play an important role in adipocyte differentiation. The strong correlation between the antidiabetic activity of TZDs and their ability to activate PPAR gamma suggests that PPAR-gamma, through downstream-regulated genes, mediates the effects of TZDs. In this report, we present the isolation and characterization of 81 genes, encoding proteins of known function, differentially expressed during TZD-stimulated differentiation of 3T3-L1 cells. By the use of different reverse- Northern blot techniques, the differential expression of 50 of these genes could be verified, and 21 genes were specifically regulated by a potent TZD during the course of adipocyte differentiation, whereas no effect of a PPAR-gamma antagonist could be observed in mature adipocytes. The differential expression of a large fraction of the isolated genes was also shown to occur in white adipose tissue of ob/ob mice treated with rosiglitazone; combined, our results suggest that an important effect of rosiglitazone in adipose tissue is based on activation of PPAR-gamma in preexisting preadipocytes found among the mature adipocytes, resulting in subsequent adipocyte differentiation. PMID- 11916925 TI - Insulin resistance, defective insulin receptor substrate 2-associated phosphatidylinositol-3' kinase activation, and impaired atypical protein kinase C (zeta/lambda) activation in myotubes from obese patients with impaired glucose tolerance. AB - Impaired glucose tolerance (IGT) is characterized by insulin resistance. Recently, defects in the insulin-signaling cascade have been implicated in the pathogenesis of insulin resistance. To study insulin signaling in IGT, we used human skeletal muscle cells in primary culture from patients with IGT and control subjects. In these cultured myotubes, we assessed insulin-induced 2-deoxyglucose uptake and early steps of the metabolic insulin-signaling cascade. Myotubes in culture from patients with IGT had insulin-induced glucose uptake that was roughly 30-50% less than that from control subjects. This insulin resistance was associated with impaired insulin receptor substrate (IRS)-2-associated phosphatidylinositol 3' (PI3) kinase activation and IRS-2 tyrosine phosphorylation as well as significantly decreased protein kinase C (PKC) zeta/lambda activation in response to insulin. IRS-1- associated PI3 kinase activation and insulin receptor autophosphorylation were comparable in the two groups. Protein expression levels for the insulin receptor, IRS-1, IRS-2, the p85 regulatory subunit of PI3 kinase, Akt, PKC-zeta/lambda, GLUT1, and GLUT4 were also similar in the two groups. In conclusion, myotubes from patients with IGT have impaired insulin-induced glucose uptake. This is associated with impaired IRS-2-associated PI3 kinase activation and PKC-zeta/lambda activation. Our results suggest that these defects may contribute to insulin resistance in IGT patients. PMID- 11916926 TI - Troglitazone downregulates delta-6 desaturase gene expression in human skeletal muscle cell cultures. AB - Delta-6 Desaturase, one of the rate-limiting enzymes, catalyzes the conversion of linoleic acid (C18:2 omega6) into gamma-linolenic acid (C18:3 omega6), arachidonic acid (C20:4 omega6), and further metabolites. Recently, it has been shown that human Delta-6 desaturase is expressed not only in liver but in a variety of human tissues, including muscle. Skeletal muscle is a major site of insulin action, and insulin sensitivity may be related to the fatty acid composition of muscle lipids. We examined the effects of troglitazone on the regulation of Delta-6 desaturase gene expression in human muscle cell cultures obtained from muscle biopsies (n = 15). Delta-6 Desaturase mRNA and peroxisome proliferator-activated receptor gamma2 (PPARgamma2) mRNA were quantified by two step RT-PCR, and the activity of the Delta-6 desaturase enzyme was estimated by gas chromatographic analysis of the omega 6-C18:3/C18:2 fatty acids ratio. In cells treated with 11.5 micromol troglitazone for 4 days, PPARgamma2 mRNA levels were significantly increased (301.0 +/- 51.5%, P < 0.05) and Delta-6 desaturase mRNA levels were significantly decreased (41.7 +/- 5.9%, P < 0.0005) compared with the untreated controls. In accordance with the decrease of Delta-6 desaturase mRNA, there was a significant decrease in the omega6-C18:3/C18:2 ratio down to 47.4 +/- 7.5% in cholesterol esters, 54.2 +/- 7.4% in phospholipids, 56.7 +/- 6.5% in nonesterified fatty acids, and 67.7 +/- 5.9% in triglycerides. The troglitazone-induced decrease in Delta-6 desaturase mRNA is associated with a change in the unsaturated fatty acid composition of the muscle cells. These results add new aspects to the known thiazolidinedione effects on lipid metabolism. PMID- 11916927 TI - Increased glucocorticoid receptor expression in human skeletal muscle cells may contribute to the pathogenesis of the metabolic syndrome. AB - Altered glucocorticoid hormone action may contribute to the etiology of the metabolic syndrome, but the molecular mechanisms are poorly defined. Tissue sensitivity to glucocorticoid is regulated by expression of the glucocorticoid receptor (GR)-alpha and 11beta-hydroxysteroid dehydrogenase type I (11beta-HSD1) mediated intracellular synthesis of active cortisol from inactive cortisone. We have analyzed GRalpha and 11beta-HSD1 expression in skeletal myoblasts from men (n = 14) with contrasting levels of insulin sensitivity (euglycemic clamp measurements of insulin-dependent glucose disposal rate), blood pressure, and adiposity. Positive associations were evident between myoblast expression of GRalpha under basal conditions and levels of insulin resistance (r(2) = 0.34, P < 0.05), BMI (r(2) = 0.49, P < 0.01), percent body fat (r(2) = 0.34, P < 0.02), and blood pressure (r(2) = 0.86, P < 0.001). Similar associations were evident when myoblasts were incubated with physiological levels of cortisol (P < 0.01 for all). Importantly, GRalpha expression was unaffected by variations in in vivo concentrations of insulin, IGF-1, or glucose concentrations. In common with the GR, 11beta-HSD1 expression in myoblasts incubated with physiological concentrations of cortisol in vitro was positively associated with levels of insulin resistance (r(2) = 0.68, P < 0.001), BMI (r(2) = 0.63, P < 0.005), and blood pressure (r(2) = 0.27, P < 0.05). Regulation of GRalpha and 11beta-HSD1 by cortisol was abolished by the GR antagonist RU38486. In summary, our data suggest that raised skeletal muscle cell expression of GRalpha and 11beta -HSD1-mediated regulation of intracellular cortisol may play a fundamental role in mechanisms contributing to the pathogenesis of the metabolic syndrome. PMID- 11916928 TI - Acute hyperglycemia induces nitrotyrosine formation and apoptosis in perfused heart from rat. AB - This study investigated coronary perfusion pressure, nitric oxide (NO) and superoxide production, nitrotyrosine (NT) formation, and cardiac cell apoptosis in isolated hearts perfused with high glucose concentration. Coronary perfusion pressure; NO and superoxide anion generation; immunostaining for NT, inducible NO synthase (iNOS), and the constitutive type of NO synthase (NOS) eNOS; iNOS and eNOS mRNA expression by Western blot and RT-PCR; and apoptosis of cardiac cells were studied in hearts perfused for 2 h with solutions containing D-glucose at a concentration of 11.1 mmol/l (control), D-glucose at the concentration of 33.3 mmol/l (high glucose), or D-glucose (33.3 mmol/l) plus glutathione (0.3 mmol/l). Perfusion of isolated hearts in conditions of high glucose concentration caused a significant increase of coronary perfusion pressure (P < 0.001) and an increase of both NO and superoxide generation. However, superoxide production was 300% higher than baseline, whereas NO production was 40% higher (P < 0.001 for both). This effect was accompanied by the formation of NT, and an increase of iNOS expression. eNOS remained unchanged. At the end of the experiments, cardiac cell apoptosis was evident in hearts perfused with high glucose. The effects of high glucose were significantly prevented by glutathione. This study demonstrates that high glucose for 2 h is enough to increase iNOS gene expression and NO release in working rat hearts. Upregulation of iNOS and raised NO generation are accompanied by a marked concomitant increase of superoxide production, a condition favoring the production of peroxynitrite, a powerful pro-oxidant that can mediate the toxic effects of high glucose on heart by itself and/or via the formation of nitrotyrosine, as suggested by the detection of cell apoptosis. PMID- 11916930 TI - Low production capacity of interleukin-10 associates with the metabolic syndrome and type 2 diabetes : the Leiden 85-Plus Study. AB - It has been suggested that the metabolic syndrome and type 2 diabetes are manifestations of the inflammatory host response. This host response is orchestrated by the production of pro- and anti-inflammatory cytokines that are under genetic control. We therefore hypothesized that a low production capacity of interleukin-10 (IL-10), a centrally operating cytokine with strong anti inflammatory properties, associates with the metabolic syndrome and type 2 diabetes in old age. In the current study, 599 inhabitants of the city of Leiden, age 85 years, were visited at their place of residence. The production capacity of the anti-inflammatory cytokine IL-10 was assessed in a whole-blood assay in which lipopolysaccharide was used as a stimulus. Serum concentrations of lipids, lipoproteins, glucose, and HbA(1c) were determined, and a history of type 2 diabetes was obtained. Serum concentrations of total cholesterol, LDL cholesterol, triglycerides, glucose, and HbA(1c) gradually decreased over strata representing higher IL-10 production capacity, whereas the concentration of HDL cholesterol gradually increased (all P < 0.01). The odds ratio for type 2 diabetes was 2.7 (95% confidence interval 1.5-4.9) when subjects with the lowest IL-10 production capacity were compared with those with the highest IL-10 production capacity. These findings showed that low IL-10 production capacity (i.e., a pro-inflammatory response) is associated with the metabolic syndrome and type 2 diabetes. PMID- 11916929 TI - A tailored therapy for the metabolic syndrome: the dual peroxisome proliferator activated receptor-alpha/gamma agonist LY465608 ameliorates insulin resistance and diabetic hyperglycemia while improving cardiovascular risk factors in preclinical models. AB - A novel nonthiazolidinedione dual peroxisome proliferator- activated receptor (PPAR)-alpha/gamma agonist, LY465608, was designed to address the major metabolic disturbances of type 2 diabetes. LY465608 altered PPAR-responsive genes in liver and fat of db/db mice and dose-dependently lowered plasma glucose in hyperglycemic male Zucker diabetic fatty (ZDF) rats, with an ED(50) for glucose normalization of 3.8 mg small middle dot kg(-1) small middle dot day(-1). Metabolic improvements were associated with enhanced insulin sensitivity, as demonstrated in female obese Zucker (fa/fa) rats using both oral glucose tolerance tests and hyperinsulinemic-euglycemic clamps. Further characterization of LY465608 revealed metabolic changes distinct from a selective PPAR-gamma agonist, which were presumably due to the concomitant PPAR-alpha agonism, lower respiratory quotient, and less fat accumulation, despite a similar impact on glycemia in male ZDF rats. In addition to these alterations in diabetic and insulin-resistant animals, LY465608 dose-dependently elevated HDL cholesterol and lowered plasma triglycerides in human apolipoprotein A-I transgenic mice, demonstrating that this compound significantly improves primary cardiovascular risk factors. Overall, these studies demonstrate that LY465608 beneficially impacts multiple facets of type 2 diabetes and associated cardiovascular risk, including those facets involved in the development of micro- and macrovascular complications, which are the major sources for morbidity and mortality in these patients. PMID- 11916931 TI - Modulation of adipose tissue expression of murine matrix metalloproteinases and their tissue inhibitors with obesity. AB - The potential role of the matrix metalloproteinase (MMP) system in the pathophysiology of the adipose tissue was investigated in a mouse model of nutritionally induced obesity. mRNA levels of 16 MMPs and 4 tissue inhibitors of MMPs (TIMPs) were measured by semiquantitative RT-PCR in adipose tissue isolated from mice maintained for 15 weeks on a standard or high-fat diet. In mice on standard diet, with the exception of MMP-8, all MMP and TIMP transcripts were detected in both gonadal and subcutaneous depots. In obese mice, the expression of MMP-3, -11, -12, -13, and -14 and TIMP-1 mRNAs was upregulated, whereas that of MMP-7, -9, -16, and -24 and TIMP-4 was downregulated. Most MMP and TIMP mRNAs were expressed at higher levels in stromal-vascular cells than in mature adipocytes. Analysis of adipose tissue by in situ fluorescent zymography revealed MMP-dependent proteolytic activities, demonstrating the presence of active MMPs in the intact tissue. In vitro conversion of adipogenic 3T3-F442A cells into mature adipocytes was associated with substantial modulations of MMP and TIMP expression. Moreover, this in vitro adipogenesis was reduced in the presence of a synthetic MMP inhibitor. Thus, the adipose tissue expresses a large array of MMPs and TIMPs, which modulate adipocyte differentiation. PMID- 11916932 TI - Decreased expression of heat shock protein 72 in skeletal muscle of patients with type 2 diabetes correlates with insulin resistance. AB - Oxidative stress has been ascribed a role in the pathogenesis of diabetes and its complications, and stress proteins have been shown to protect organisms in vitro and in vivo against oxidative stress. To study the putative role of one of the most abundant cytoprotective stress proteins, inducible cytoplasmic 72-kDa-mass heat shock protein (Hsp-72), in the pathogenesis of diabetes, we measured its mRNA concentration in muscle biopsies from six type 2 diabetic patients and six healthy control subjects (protocol 1) as well as in 12 twin pairs discordant for type 2 diabetes and 12 control subjects undergoing a euglycemic-hyperinsulinemic clamp in combination with indirect calorimetry (protocol 2). The amount of Hsp-72 mRNA in muscle was significantly lower in type 2 diabetic patients than in healthy control subjects (in protocol 1: 5.2 +/- 2.2 vs. 53 +/- 32 million copies of Hsp-72 mRNA/microg total RNA, n = 6, P = 0.0039; in protocol 2: 3.2 +/- 3.3 vs. 43 +/- 31 million copies of Hsp-72 mRNA/microg total RNA, n = 12, P = 0.0001). Hsp-72 mRNA levels were also markedly reduced in the nondiabetic co twins compared with healthy control subjects (5.8 +/- 5.0 vs. 43 +/- 31, n = 12, P = 0.0001), but they were also statistically significantly different from their diabetic co-twins when the difference between the pairs was compared (P = 0.0280). Heat shock protein mRNA content in muscle of examined patients correlated with the rate of glucose uptake and other measures of insulin stimulated carbohydrate and lipid metabolism. In conclusion, the finding of decreased levels of Hsp-72 mRNA in skeletal muscle of patients with type 2 diabetes and its relationship with insulin resistance raises the question of whether heat shock proteins are involved in the pathogenesis of skeletal muscle insulin resistance in type 2 diabetes. PMID- 11916933 TI - Thiazolidinedione treatment normalizes insulin resistance and ischemic injury in the zucker Fatty rat heart. AB - Obesity is associated with risk factors for cardiovascular disease, including insulin resistance, and can lead to cardiac hypertrophy and congestive heart failure. Here, we used the insulin-sensitizing agent rosiglitazone to investigate the cellular mechanisms linking insulin resistance in the obese Zucker rat heart with increased susceptibility to ischemic injury. Rats were treated for 7 or 14 days with 3 mg/kg per os rosiglitazone. Hearts were isolated and perfused before and during insulin stimulation or during 32 min low-flow ischemia at 0.3 ml small middle dot min(-1) small middle dot grams wet wt(-1) and reperfusion. D[2 (3)H]glucose was used as a tracer of glucose uptake, and phosphorus-31 nuclear magnetic resonance spectroscopy was used to follow energetics during ischemia. At 12 months of age, obese rat hearts were insulin resistant with decreased GLUT4 protein expression. During ischemia, glucose uptake was lower and depletion of ATP was greater in obese rat hearts, thereby significantly impairing recovery of contractile function during reperfusion. Rosiglitazone treatment normalized the insulin resistance and restored GLUT4 protein levels in obese rat hearts. Glucose uptake during ischemia was also normalized by rosiglitazone treatment, thereby preventing the greater loss of ATP and restoring recovery of contractile function to that of lean rat hearts. We conclude that rosiglitazone treatment, by normalizing glucose uptake, protected obese rat hearts from ischemic injury. PMID- 11916934 TI - Xanthine oxidase is involved in free radical production in type 1 diabetes: protection by allopurinol. AB - The aim of this work was to study the mechanism of free radical formation in type 1 diabetes and its possible prevention. We have found oxidation of blood glutathione and an increase in plasma lipoperoxide levels in both human type 1 diabetes and experimental diabetes. Peroxide production by mitochondria does not increase in diabetes. On the contrary, the activity of xanthine oxidase, a superoxide-generating enzyme, increases in liver and plasma of diabetic animals. The increase in plasma xanthine oxidase activity may be explained by the increase in the hepatic release of this enzyme, which is not due to nonspecific membrane damage: release of other hepatic enzymes, such as the amino transferases, does not increase in diabetes. Superoxide formation by aortic rings of rabbits increases significantly in diabetes. This is completely inhibited by allopurinol, an inhibitor of xanthine oxidase. Heparin, which releases xanthine oxidase from the vessel wall, also decreases superoxide formation by aortic rings of diabetic animals. Treatment with allopurinol decreases oxidative stress in type 1 diabetic patients: hemoglobin glycation, glutathione oxidation, and the increase in lipid peroxidation are prevented. These results may have clinical significance in the prevention of late-onset vascular complications of diabetes. PMID- 11916935 TI - Dose-dependent vasodilating effects of insulin on adenosine-stimulated myocardial blood flow. AB - In the peripheral vasculature, insulin induces time- and dose-dependent vasodilation. We have recently demonstrated that insulin potentiates adenosine stimulated myocardial blood flow. However, it is unknown whether insulin's effects on the coronary vasculature are dose dependent. In this study, we quantitated myocardial blood flow and adenosine-stimulated coronary flow (140 microg.kg(-1).min(-1) for 5 min) in 10 healthy men (age, 32 +/- 6 years; BMI, 24.1 +/- 1.8 kg/m(2)) using positron emission tomography and (15)O-labeled water. Hyperemic myocardial blood flow was measured in the basal state, during euglycemic physiological hyperinsulinemia (serum insulin approximately 65 mU/l) and during supraphysiological hyperinsulinemia (serum insulin approximately 460 mU/l). Basal myocardial blood flow was 0.84 +/- 0.17 ml.g(-1).min(-1). Physiological hyperinsulinemia increased the adenosine-stimulated flow by 20% (from 3.92 +/- 1.17 to 4.72 +/- 0.96 ml.g(- 1).min(-1); P < 0.05). Supraphysiological hyperinsulinemia further enhanced the adenosine-stimulated flow by 19% (to 5.61 +/- 1.03 ml.g(-1).min(-1); P < 0.05). These effects were not explained by changes in systemic hemodynamics, since coronary resistance decreased during each insulin infusion (P < 0.05). In addition, hyperemic myocardial blood flow responses during insulin stimulation were positively correlated with whole-body glucose uptake. The results demonstrate that insulin is able to enhance hyperemic myocardial blood flow in a dose-dependent manner in healthy subjects. These effects might contribute to the known beneficial dose dependent effects of insulin on myocardial ischemia. PMID- 11916936 TI - Elevated levels of acute-phase proteins and plasminogen activator inhibitor-1 predict the development of type 2 diabetes: the insulin resistance atherosclerosis study. AB - Elevated serum levels of acute-phase proteins, indicating chronic subclinical inflammation, have been associated with cardiovascular disease as well as the insulin resistance syndrome. Chronic inflammation may also be a risk factor for developing type 2 diabetes. We studied the concentrations of C-reactive protein (CRP), fibrinogen, and plasminogen activator inhibitor-1 (PAI-1) in 1,047 nondiabetic subjects in relation to incident diabetes within 5 years in the Insulin Resistance Atherosclerosis Study. Subjects with diabetes at follow-up (n = 144) had higher baseline levels of fibrinogen (mean +/- SD; 287.8 +/- 58.8 vs. 275.1 +/- 56.0 mg/dl; P = 0.013) as well as of CRP (median [interquartile range]; 2.40 [1.29, 5.87] vs. 1.67 mg/l [0.75, 3.41]; P = 0.0001) and PAI-1 (24 [15, 37.5] vs. 16 ng/ml [9, 27]; P = 0.0001) than nonconverters. The odds ratio (OR) of converting to diabetes was significantly increased with increasing baseline concentrations of the inflammatory markers. In contrast to PAI-1, the association of CRP and fibrinogen with incident diabetes was significantly attenuated after adjustment for body fat (BMI or waist circumference) or insulin sensitivity (S(I)), as assessed by a frequently sampled intravenous glucose tolerance test. In a logistic regression model that included age, sex, ethnicity, clinical center, smoking, BMI, S(I), physical activity, and family history of diabetes, PAI-1 still remained significantly related to incident type 2 diabetes (OR [95% CI] for 1 SD increase: 1.61 [1.20-2.16]; P = 0.002). Chronic inflammation emerges as a new risk factor for the development of type 2 diabetes; PAI-1 predicts type 2 diabetes independent of insulin resistance and other known risk factors for diabetes. PMID- 11916937 TI - Lipid infusion impairs physiologic insulin-mediated capillary recruitment and muscle glucose uptake in vivo. AB - Infusion of triglycerides and heparin causes insulin resistance in muscle. Because the vascular actions of insulin, particularly capillary recruitment, may contribute to the increase in glucose uptake by skeletal muscle, we investigated the effects of Intralipid/heparin infusion on the hemodynamic actions of insulin during clamp conditions. Saline or 10% Intralipid/heparin (33 U/ml) was infused into anesthetized rats at 20 microl/min for 6 h. At 4 h into the saline infusion, a 2-h hyperinsulinemic (3 mU. min(-1).kg(-1))-euglycemic clamp was conducted (Ins group). At 4 h into the lipid infusion, a 2-h saline control (Lip group) or 2-h hyperinsulinemic-euglycemic clamp (Lip + Ins group) was conducted. Arterial blood pressure, heart rate, femoral blood flow (FBF), hindleg vascular resistance, glucose infusion rate (GIR), hindleg glucose uptake (HGU), and muscle 2 deoxyglucose uptake (R'g) were measured. Capillary recruitment, as measured by metabolism of infused 1-methylxanthine (1-MX), was also assessed. When compared with either Lip or Lip + Ins, Ins had no effect on arterial blood pressure, heart rate, FBF, or vascular resistance but increased GIR, HGU, and R'g of soleus, plantaris, extensor digitorum longus, and gastrocnemius red muscles and hindlimb 1-MX metabolism. GIR, HGU, and R'g of soleus, plantaris, gastrocnemius red, and the combined muscles and 1-MX metabolism were less in Lip + Ins than in Ins rats. HGU correlated closely with hindleg capillary recruitment (r = 0.86, P < 0.001) but not total hindleg blood flow. In conclusion, acute elevation of plasma free fatty acids blocks insulin-mediated glucose uptake and capillary recruitment. PMID- 11916938 TI - Flux through the hexosamine pathway is a determinant of nuclear factor kappaB- dependent promoter activation. AB - The hexosamine pathway may mediate some of the toxic effects of glucose. We hypothesized that flux through this pathway might regulate the activity of nuclear factor kappaB (NF-kappaB)-dependent genes in mesangial cells (MCs). In MCs, RT-PCR revealed that high glucose (30 mmol/l) and glucosamine (1 mmol/l) increased mRNA levels for vascular cell adhesion molecule 1 (VCAM-1) and increased the activity of an NF-kappaB enhancer by 1.5- and 2-fold, respectively. Overexpression of glutamine:fructose-6-phosphate amidotransferase (GFAT), the rate-limiting enzyme for flux through the hexosamine pathway, led to a 2.2-fold increase in NF-kappaB enhancer activity; the combination of GFAT overexpression and high glucose increased activity 2.8-fold, and these increases were prevented by 40 micromol/l O-diazoacetyl-L-serine (azaserine) or 6-diazo-5-oxonorleucine. High glucose, glucosamine, and GFAT overexpression increased binding of MC nuclear proteins to NF-kappaB consensus sequences. Immunoblotting revealed that the p65 subunit of NF-kappaB was O-glycosylated in MC cultured in physiologic glucose and that significant enhancement occurred with high glucose and glucosamine. Both glucose and glucosamine dose-dependently increased human VCAM-1 promoter activity. In addition, GFAT overexpression activated the VCAM-1 promoter (2.25-fold), with further augmentation by high glucose and abrogation by inhibitors of GFAT, NF-kappaB, and O-glycosylation. Inactivation of the two NF kappaB sites in the VCAM-1 promoter abolished its response to high glucose, glucosamine, and GFAT overexpression. These results suggest that increased flux through the hexosamine pathway leads to NF-kappaB-dependent promoter activation in MCs. PMID- 11916939 TI - Increased urinary albumin excretion, endothelial dysfunction, and chronic low grade inflammation in type 2 diabetes: progressive, interrelated, and independently associated with risk of death. AB - In 328 type 2 diabetic patients followed for 9.0 years (mean), we investigated whether endothelial dysfunction and chronic inflammation (estimated from plasma markers) can explain the association between (micro)albuminuria and mortality. Of the patients, 113 died. Mortality was increased in patients with baseline microalbuminuria or macroalbuminuria (odds ratios as compared with normoalbuminuria, 1.78 [P < 0.05] and 2.86 [P < 0.01]) and in patients with soluble vascular cell adhesion molecule 1 in the third tertile and C-reactive protein in the second and third tertiles (odds ratios as compared with the first tertile, 2.05 [ P < 0.01], and 1.80 [P < 0.05] and 2.92 [ P < 0.01]). These associations were mutually independent. The mean yearly change in urinary albumin excretion was 9.4%; in von Willebrand factor, 8.1%; in tissue-type plasminogen activator, 2.8%; in soluble vascular cell adhesion molecule 1, 5.2%; in soluble E selectin, -2.3%; in C-reactive protein, 3.8%; and in fibrinogen, 2.3%. The longitudinal development of urinary albumin excretion was significantly and independently determined by baseline levels of and the longitudinal development of BMI, systolic blood pressure, serum creatinine, glycated hemoglobin and plasma von Willebrand factor (baseline only), soluble E-selectin (baseline only), tissue type plasminogen activator, C-reactive protein, and fibrinogen. The longitudinal developments of markers of endothelial function and inflammation were interrelated. In type 2 diabetes, increased urinary albumin excretion, endothelial dysfunction, and chronic inflammation are interrelated processes that develop in parallel, progress with time, and are strongly and independently associated with risk of death. PMID- 11916940 TI - Overexpression of the sarcoplasmic reticulum Ca(2+)-ATPase improves myocardial contractility in diabetic cardiomyopathy. AB - Diabetic cardiomyopathy is characterized by reduced cardiac contractility due to direct changes in heart muscle function independent of vascular disease. An important contributor to contractile dysfunction in the diabetic state is an impaired sarcoplasmic reticulum (SR) function, leading to disturbed intracellular calcium handling. We investigated whether overexpression of the SR calcium pump (SERCA2a) in transgenic mice could reduce the impact of diabetes on the development of cardiomyopathy. Diabetes was induced by streptozotocin injection (200 mg/kg), and left ventricular (LV) function was analyzed in isolated hearts 3 weeks later. In diabetic hearts systolic LV pressure was decreased by 15% and maximum speed of relaxation (-dP/dt) by 34%. Functional changes were also assessed in isolated papillary muscles. Active force was reduced by 61% and maximum speed of relaxation by 65% in the diabetic state. The contractile impairment was accompanied by a 30% decrease in SERCA2a protein in diabetic mice. We investigated whether increased SERCA2a expression in transgenic SERCA2a overexpressing mice could compensate for the diabetes-induced decrease in cardiac function. Under normal conditions, SERCA2a overexpressors show improved contractile performance relative to wild-type (WT) mice (-dP/dt: 3,169 vs. 2,559 mmHg/s, respectively). Measurement of LV function in hearts from diabetic SERCA2a mice revealed systolic and diastolic functions that were similar to WT control mice and markedly improved relative to diabetic WT mice (-dP/dt: 2,534 vs. 1,690 mmHg/s in diabetic SERCA2a vs. diabetic WT mice, respectively). Similarly, the contractile behavior of isolated papillary muscles from diabetic SERCA2a mice was not different from that of control mice. SERCA2a protein expression was higher (60%) in diabetic SERCA2a mice than WT diabetic mice. These results indicate that overexpression of SERCA2a can protect diabetic hearts from severe contractile dysfunction, presumably by improving the calcium sequestration of the SR. PMID- 11916942 TI - Homocysteine induces protein kinase C activation and stimulates c-Fos and lipoprotein lipase expression in macrophages. AB - Hyperhomocysteinemia is an independent risk factor for cardiovascular disease in human diabetes. Among the multiple factors that may account for the atherogenicity of homocysteine (Hcys) in patients with diabetes, macrophage (Mo) lipoprotein lipase (LPL) has unique features in that it is increased in human diabetes and acts as a proatherogenic factor in the arterial wall. In the present study, we determined the direct regulatory effect of Hcys on Mo LPL gene expression and secretion. Incubation of J774 Mo with Hcys increased, in a time- and dose-dependent manner, LPL mRNA expression and secretion. Induction of LPL gene expression was biphasic, peaking at 1 and 6 h. Whereas Hcys treatment increased protein kinase C (PKC) activity in Mo, pretreatment of Mo with PKC inhibitors totally suppressed Hcys-induced LPL mRNA expression. Hcys also increases the levels of c-fos mRNA in Mo and enhanced nuclear protein binding to the AP-1 sequence of the LPL gene promoter. Overall, these results demonstrate that Hcys stimulates Mo LPL at both the gene and protein levels and that Hcys induced LPL mRNA expression requires PKC activation. They also suggest a possible role of c-fos in the stimulatory effect of Hcys on Mo LPL mRNA expression. These observations suggest a new mechanism by which Hcys may exert its proatherogenic effects in human diabetes. PMID- 11916941 TI - Caspase activation in retinas of diabetic and galactosemic mice and diabetic patients. AB - Apoptosis of retinal capillary cells begins early in diabetes and likely contributes to the capillary obliteration that is an important feature of diabetic retinopathy. Caspases are proteolytic enzymes that are closely involved in the induction and execution phases of apoptosis, but their role in the development of diabetic retinopathy has not been studied previously. Our study focused on the measurement of activities of multiple caspases in retinas of mice at different durations of diabetes. Several caspases (including caspases-1, -2, 6, -8, and -9) were activated as early as 2 months of diabetes. The caspases activity pattern changed with increasing duration of disease, suggesting a slowly developing caspases cascade. Activities of executioner caspases (e.g., cas-6 and 3) became elevated after longer duration of diabetes, and the induction of cas-3 activity was associated with the duration of diabetes at which capillary cells begin to show evidence of undergoing apoptosis. Retinas from patients with type 2 diabetes likewise showed a significant increase in activities of cas-1, -3, -4, and -6. For comparison, retinal caspases were also measured in experimental galactosemia, another model that develops a diabetic-like retinopathy. The pattern of caspases activation differed between diabetes and galactosemia, but cas-1 activity became elevated soon after elevation of blood hexose concentration in both. Caspases offer new therapeutic targets to test the role of apoptosis in the development of diabetic retinopathy. PMID- 11916943 TI - Polymorphism in ecto-nucleotide pyrophosphatase/phosphodiesterase 1 gene (ENPP1/PC-1) and early development of advanced diabetic nephropathy in type 1 diabetes. AB - A polymorphism in the ecto-nucleotide pyrophosphatase/phosphodiesterase 1 gene (ENPP1) (previously known as PC-1), resulting in an amino acid change from lysine to glutamine at codon 121 (K121Q), is associated with insulin resistance. A small follow-up study of patients with type 1 diabetes and proteinuria found that renal function declines more rapidly in carriers of the Q variant than in noncarriers. To examine this finding further, we conducted a large case-control study and a family-based study. Genomic DNA was obtained from 659 patients: 307 with normal urinary albumin excretion despite diabetes duration of >15 years (control subjects) and 352 with advanced diabetic nephropathy, of whom 200 had persistent proteinuria and 152 had end-stage renal disease (ESRD). Individuals were genotyped for Q and K variants using a previously described protocol. The frequency of Q variant carriers was 21.5% in control subjects, 31.5% in subjects with proteinuria, and 32.2% in subjects with ESRD (P = 0.012). In a stratified analysis according to duration of diabetes, the risk of early-onset ESRD for carriers of the Q variant was 2.3 times that for noncarriers (95% CI, 1.2-4.6). The Q variant was not associated with late-onset ESRD. Similar findings were obtained in a family-based study. We conclude that carriers of the Q variant of ENPP1 are at increased risk for developing ESRD early in the course of type 1 diabetes. PMID- 11916945 TI - ATP-dependent K(+) channels contribute to local metabolic coronary vasodilation in experimental diabetes. AB - This study tested whether ATP-dependent K(+) channels (K(ATP) channels) are an important mechanism of functional coronary hyperemia in conscious, instrument implanted diabetic dogs. Data were collected at rest and during exercise before and after induction of diabetes with alloxan monohydrate (40-60 mg/kg intravenously). K(ATP) channels were inhibited with glibenclamide (1 mg/kg intravenously). In nondiabetic dogs, arterial plasma glucose concentration increased from 4.8 +/- 0.3 to 21.5 +/- 2.2 mmol/l 1 week after alloxan injection. In nondiabetic dogs, exercise increased myocardial oxygen consumption (MVO(2)) 3.4-fold, myocardial O(2) delivery 3.0-fold, and heart rate 2.4-fold. Coronary venous PO(2) decreased from 19.9 +/- 0.8 mmHg at rest to 14.8 +/- 0.8 mmHg during exercise. Diabetes significantly reduced myocardial O(2) delivery and lowered coronary venous PO(2) from 16.3 +/- 0.6 mmHg at rest to 13.1 +/- 0.9 mmHg during exercise. Glibenclamide did not alter the slope of the coronary venous PO(2) versus MVO(2) relationship in nondiabetic dogs. In diabetic dogs, however, glibenclamide further reduced myocardial O(2) delivery; coronary venous PO(2) fell to 9.0 +/- 1.0 mmHg during exercise, and the slope of the coronary venous PO(2) versus MVO(2) relationship steepened. These findings indicate that K(ATP) channels contribute to local metabolic coronary vasodilation in alloxan-induced diabetic dogs. PMID- 11916944 TI - Altered phosphodiesterase 3-mediated cAMP hydrolysis contributes to a hypermotile phenotype in obese JCR:LA-cp rat aortic vascular smooth muscle cells: implications for diabetes-associated cardiovascular disease. AB - Cardiovascular diseases represent a significant cause of morbidity and mortality in diabetes. Of the many animal models used in the study of non-insulin-dependent (type 2) diabetes, the JCR:LA-cp rat is unique in that it develops insulin resistance in the presence of obesity and manifests both peripheral and coronary vasculopathies. In this animal model, arterial vascular smooth muscle cells (VSMCs) from homozygous obese (cp/cp) rats, but not from age-matched healthy (+/+ or + /cp, collectively defined +/?) littermates, display an " activated" phenotype in vitro and in vivo and have an elevated level of cAMP phosphodiesterase (PDE) activity. In this report, we confirm that cp/cp rat aortic VSMCs have an elevated level of PDE3 activity and show that only particulate PDE3 (PDE3B) activity is elevated. In marked contrast to results obtained in + /? VSMCs, simultaneous activation of adenylyl cyclase and inhibition of PDE3 activity in cp/cp VSMCs synergistically increased cAMP. Although PDE3 inhibition did not potentiate the antimigratory effects of forskolin on +/? VSMCs, PDE3 inhibition did markedly potentiate the forskolin induced inhibition of migration of cp/cp-derived VSMCs. Although PDE3 activity was elevated in cp/cp rat aortic VSMCs, levels of expression of cytosolic PDE3 (PDE3A) and PDE3B in +/? and cp/cp VSMCs, as well as activation of these enzymes following activation of the cAMP-protein kinase A signaling cascade, were not different. Our data are consistent with an increased role for PDE3 in regulating cAMP-dependent signaling in cp/cp VSMCs and identify PDE3 as a cellular activity potentially responsible for the phenotype of cp/cp VSMCs. PMID- 11916947 TI - Early decrease of skin blood flow in response to locally applied pressure in diabetic subjects. AB - Pressure ulcers are common debilitating complications of diabetes that are caused by tissue ischemia. Skin blood flow in response to locally applied pressure might be impaired in diabetic patients because of the combined effects of a typically low skin temperature and alterations in microcirculatory function, and could be worsened by neuropathy. We measured skin blood flow by laser Doppler flowmetry over the internal anklebone in response to local pressure applied at 5.0 mmHg/min in three groups of diabetic patients (with clinical and subclinical neuropathy and without neuropathy) and in healthy matched control subjects at usual room temperature. Compared with in matched control subjects with comparable skin temperatures (29.3 +/- 0.4 vs. 28.7 +/- 0.4 degrees C), in diabetic patients the skin blood flow response to locally applied pressure was further impeded, even in those without neuropathy. Indeed, skin blood flow decreased significantly from baseline at much lower applied pressure (7.5 mmHg) in diabetic subjects, again even in those without neuropathy, than in control subjects (48.8 mmHg). The large difference between these pressures could partially explain diabetic patients' high risk of developing decubitus and plantar ulcers. PMID- 11916946 TI - A common promoter polymorphism in the hepatic lipase gene (LIPC-480C>T) is associated with an increase in coronary calcification in type 1 diabetes. AB - Type 1 diabetes is associated with coronary heart disease (CHD) and coronary artery calcification (CAC), a measure of subclinical CHD. The hepatic lipase gene promoter polymorphism (LIPC-480C>T) is a common variant affecting lipid metabolism. This study examined the relation between the LIPC-480C>T and CAC in type 1 diabetes. In the type 1 diabetic patients studied, 56% had CAC >0 Agatston units (AU). These subjects had a longer duration of diabetes (26.2 +/- 1.3 vs. 17.8 +/- 1.4 years; P < 0.001), lower HDL cholesterol levels (55.7 +/- 2.4 vs. 61.0 +/- 2.5 mg/dl; P = 0.05), higher triglyceride levels (101 +/- 17.3 vs. 66 +/ 7.6 mg/dl; P < 0.05), and higher diastolic blood pressure (79.7 +/- 1.0 vs. 76.0 +/- 1.4 mmHg; P < 0.05). The LIPC-480 T allele was more common in subjects with CAC (frequency = 0.31 +/- 0.05 vs. 0.14 +/- 0.04; P = 0.006). The proportion with CAC was 44% in LIPC-480CC subjects, 71% in heterozygotes, and 83% in LIPC-480TT subjects (P < 0.01). LIPC-480 T allele frequency increased as the amount of CAC increased (P = 0.007). LIPC-480 genotype was independently associated with the CAC (odds ratio = 2.90, 95% CI 1.22-6.92, P < 0.05) after adjusting for duration of diabetes, age, sex, diastolic blood pressure, HDL cholesterol, and triglyceride levels. In conclusion, the LIPC-480C>T polymorphism was associated with subclinical CHD in type 1 diabetes. This genetic variant may identify subjects in which early intervention to prevent CHD may be appropriate. PMID- 11916948 TI - Difference in ischemic regulation of vascular endothelial growth factor and pigment epithelium--derived factor in brown norway and sprague dawley rats contributing to different susceptibilities to retinal neovascularization. AB - The present study compared susceptibilities of Sprague Dawley (SD) and Brown Norway (BN) rats with ischemia-induced retinal neovascularization. An exposure to constant hyperoxia followed by normoxia induced significant retinal neovascularization in BN rats but not in SD rats, as demonstrated by fluorescein retinal angiography, measurement of avascular area, and count of preretinal vascular cells. These results indicate a rat strain difference in susceptibility to retinal neovascularization. To understand the molecular basis responsible for the strain difference, we have measured the levels of pigment epithelium-derived factor (PEDF), an angiogenic inhibitor, and vascular endothelial growth factor (VEGF), a major angiogenic stimulator in the retina. The hyperoxia-treated BN rats showed a significant reduction in retinal PEDF, but they showed a substantial increase of VEGF at both the protein and RNA levels, resulting in an increased VEGF-to-PEDF ratio. Hyperoxia-treated SD rats showed changes in PEDF and VEGF levels that were less in magnitude and of shorter duration than in BN rats. In age-matched normal BN and SD rats, however, there was no detectable difference in the basal VEGF-to-PEDF ratio between the strains. These observations support the idea that different regulation of angiogenic inhibitors and stimulators under ischemia are responsible for the differences in susceptibility to ischemia-induced retinal neovascularization in SD and BN rats. PMID- 11916949 TI - Abnormalities of glucose metabolism in patients with early renal failure. AB - Abnormalities of glucose metabolism and hyperinsulinemia have been demonstrated in patients with end-stage renal disease and may contribute to the development of atherosclerotic complications in these patients. In the present study, we investigated the stage of renal failure in which abnormalities of glucose metabolism develop and whether these abnormalities were associated with an increased prevalence of cardiovascular events in patients with early renal failure. In 321 untreated essential hypertensive patients, we assessed renal function by measuring 24-h creatinine clearance, urinary protein excretion, and microalbuminuria; we assessed cardiovascular status by clinical and laboratory tests; and we measured plasma glucose, insulin, and C-peptide levels at fasting and after a 75-g oral glucose load. To evaluate insulin sensitivity, a hyperinsulinemic-euglycemic clamp was performed in a subgroup of 104 patients. Patients with creatinine clearance < 30 ml.min(-1).1.73 m(-2), severe hypertension, BMI < 30 kg/m(2), and diabetes or family history of diabetes were excluded. Hypertensive patients were found to be hyperinsulinemic when compared with 92 matched normotensive subjects. Early renal failure (creatinine clearance < 90 ml.min(-1).1.73 m(-2)) caused by hypertensive nephrosclerosis was detected in 116 of 321 patients. Analysis of patients with varying degrees of renal function impairment demonstrated increased plasma glucose and insulin response to oral glucose load, decreased fasting glucose-to-insulin ratio, and reduced sensitivity to insulin only in those patients with creatinine clearance < 50 ml.min(-1).1.73 m(-2). Parameters of glucose metabolism were not correlated with creatinine clearance and microalbuminuria. Prevalence of atherosclerotic cardiovascular events was significantly related to reduction of creatinine clearance, but parameters of glucose metabolism were comparable in patients with and without evidence of atherosclerotic damage. Thus, in patients with hypertensive nephrosclerosis and early impairment of glomerular filtration, alterations of glucose metabolism become evident only when creatinine clearance is < 50 ml.min(-1).1.73 m(-2) and are not related to microalbuminuria and cardiovascular complications. PMID- 11916950 TI - Hepatic expression of microsomal triglyceride transfer protein and in vivo secretion of triglyceride-rich lipoproteins are increased in obese diabetic mice. AB - Secondary hyperlipidemia is a major cardiovascular risk factor in individuals with type 2 diabetes. Increased hepatic production of apolipoprotein B (apoB) containing lipoproteins contributes to the elevated plasma levels, but the mechanism is poorly understood. Recent results have established that microsomal triglyceride transfer protein (MTP) is rate limiting for the assembly and secretion of apoB-containing lipoproteins. To better understand the mechanism of type 2 diabetes-associated hyperlipidemia, we quantified hepatic MTP mRNA levels, hepatic microsomal triglyceride transfer activity, and in vivo triglyceride secretion from the liver in two diabetic mouse models. Obese diabetic (ob/ob) mice had 45% higher (P = 0.006) hepatic MTP mRNA levels, 54% higher (P < 0.0001) microsomal triglyceride transfer activity, and 70% higher (P < 0.0001) in vivo triglyceride secretion rates compared with ob/+ control mice. In contrast, in lean streptozotocin-treated diabetic mice, hepatic MTP mRNA levels were unchanged, whereas microsomal triglyceride transfer activity and in vivo triglyceride secretion rates were marginally decreased. These studies suggest that obesity-induced type 2 diabetes in mice confers increases in hepatic MTP expression and secretion of triglyceride-rich lipoproteins. High blood glucose and altered hepatic expression of sterol regulatory element binding protein genes play a minor role in this diabetic response. PMID- 11916951 TI - The second activating glucokinase mutation (A456V): implications for glucose homeostasis and diabetes therapy. AB - In this study, a second case of hyperinsulinemic hypoglycemia due to activation of glucokinase is reported. The 14-year-old proband had a history of neonatal hypoglycemia, treated with diazoxide. He was admitted with coma and convulsions due to nonketotic hypoglycemia. His BMI was 34 kg/m(2), and his fasting blood glucose ranged from 2.1 to 2.7 mmol/l, associated with inappropriately high serum levels of insulin, C-peptide, and proinsulin. An oral glucose tolerance test (OGTT) showed exaggerated responses of these peptides followed by profound hypoglycemia. Treatment with diazoxide and chlorothiazide was effective. His mother never had clinical hypoglycemic symptoms, even though her fasting blood glucose ranged from 2.9 to 3.5 mmol/l. Increases in serum insulin, C-peptide, and proinsulin in response to an OGTT suggested a lower threshold for glucose stimulated insulin release (GSIR). Screening for mutations in candidate genes revealed a heterozygous glucokinase mutation in exon 10, substituting valine for alanine at codon 456 (A456V) in the proband and his mother. The purified recombinant glutathionyl S-transferase fusion protein of the A456V glucokinase revealed a decreased glucose S(0.5) (the concentration of glucose needed to achieve the half-maximal rate of phosphorylation) from 8.04 (wild-type) to 2.53 mmol/l. The mutant's Hill coefficient was decreased, and its maximal specific activity k(cat) was increased. Mathematical modeling predicted a markedly lowered GSIR threshold of 1.5 mmol/l. The theoretical and practical implications are manifold and significant. PMID- 11916952 TI - Genome-wide search for type 2 diabetes in Japanese affected sib-pairs confirms susceptibility genes on 3q, 15q, and 20q and identifies two new candidate Loci on 7p and 11p. AB - The genetic background that predisposes the Japanese population to type 2 diabetes is largely unknown. Therefore, we conducted a 10-cM genome-wide scan for type 2 diabetes traits in the 359 affected individuals from 159 families, yielding 224 affected sib-pairs of Japanese origin. Nonparametric multipoint linkage analyses performed in the whole population showed one suggestive linked region on 11p13-p12 (maximum logarithm of odds score [MLS] 3.08, near Pax6) and seven potentially linked regions (MLS >1.17) at 1p36-p32, 2q34, 3q26-q28, 6p23, 7p22-p21, 15q13-q21, and 20q12-q13 (near the gene for hepatocyte nuclear factor 4alpha [HNF-4alpha]). Subset analyses according to maximal BMI and early age at diagnosis added suggestive evidence of linkage with type 2 diabetes at 7p22-p21 (MLS 3.51), 15q13-q21 (MLS 3.91), and 20q12-q13 (MLS 2.32). These results support previous indication for linkage found on chromosome 3q, 15q, and 20q in other populations and identifies two new potential loci on 7p and 11p that may confer genetic risk for type 2 diabetes in the Japanese population. PMID- 11916953 TI - Type 2 diabetes, APOE gene, and the risk for dementia and related pathologies: The Honolulu-Asia Aging Study. AB - Type 2 diabetes may be a risk factor for dementia, but the associated pathological mechanisms remains unclear. We evaluated the association of diabetes alone or combined with the apolipoprotein E (APOE) gene with incident dementia and neuropathological outcomes in a population-based cohort of 2,574 Japanese American men enrolled in the Honolulu-Asia Aging Study, including 216 subjects who underwent autopsy. Type 2 diabetes was ascertained by interview and direct glucose testing. Dementia was assessed in 1991 and 1994 by clinical examination and magnetic resonance imaging and was diagnosed according to international guidelines. Logistic regression was used to assess the RR of developing dementia, and log-linear regression was used to estimate the incident rate ratio (IRR) of neuropathological outcomes. Diabetes was associated with total dementia (RR 1.5 [95% CI 1.01-2.2]), Alzheimer's disease (AD; 1.8 [1.1-2.9]), and vascular dementia (VsD; 2.3 [1.1-5.0]). Individuals with both type 2 diabetes and the APOE epsilon4 allele had an RR of 5.5 (CI 2.2-13.7) for AD compared with those with neither risk factor. Participants with type 2 diabetes and the epsilon4 allele had a higher number of hippocampal neuritic plaques (IRR 3.0 [CI 1.2-7.3]) and neurofibrillary tangles in the cortex (IRR 3.5 [1.6-7.5]) and hippocampus (IRR 2.5 [1.5-3.7]), and they had a higher risk of cerebral amyloid angiopathy (RR 6.6, 1.5-29.6). Type 2 diabetes is a risk factor for AD and VsD. The association between diabetes and AD is particularly strong among carriers of the APOE epsilon4 allele. The neuropathological data are consistent with the clinical results. PMID- 11916954 TI - Increased proinsulin levels and decreased acute insulin response independently predict the incidence of type 2 diabetes in the insulin resistance atherosclerosis study. AB - Previous studies have indicated that beta-cell dysfunction predicts the development of diabetes, although it is unknown whether the use of combinations of insulin secretory measures further improves prediction. The Insulin Resistance Atherosclerosis Study is a prospective, multicenter, epidemiological study of the relationship between insulin sensitivity and the risk of diabetes and cardiovascular disease. At baseline, fasting concentrations of insulin, intact proinsulin (PI), and split PI were measured, and acute insulin response (AIR) was determined during a frequently sampled intravenous glucose tolerance test (FSIGTT). Subjects who were nondiabetic at baseline (n = 903) were reexamined after 5 years of follow-up; 148 had developed diabetes. In separate logistic regression models adjusted for age, sex, clinic, and ethnicity, 1 SD differences in measures of beta-cell dysfunction were associated with diabetes incidence (AIR: odds ratio [OR] 0.37, 95% CI 0.27-0.52; intact PI: OR 1.90, 95% CI 1.57 2.30; split PI: OR 1.94, 95% CI 1.63-2.31). After additional adjustment for BMI, impaired glucose tolerance, and insulin sensitivity, these measures continued to be significantly associated with risk of diabetes (all P < 0.0001). Furthermore, in models that included both PI and AIR, each was an independent predictor, and individuals who had combined low AIR and high PI experienced the highest diabetes risk. In conclusion, both low AIR and high PI independently predicted diabetes in a well-characterized multiethnic population. Although fasting PI is simpler to assess, determining AIR from an FSIGTT may further improve prediction. If pharmacological agents to prevent diabetes are proved to be efficacious in ongoing clinical trials, then it may be beneficial to perform FSIGTTs to identify better (for intensive intervention) prediabetic subjects who would ultimately require lifelong pharmacological therapy. PMID- 11916956 TI - Peroxisome proliferator-activated receptor-gamma coactivator-1 gene locus: associations with obesity indices in middle-aged women. AB - Peroxisome proliferator-activated receptor-gamma coactivator-1 (PPARGC1) is a transcriptional coactivator that has been implicated in the regulation of genes involved in energy metabolism. We studied associations of two polymorphisms identified in PPARGC1 transcripts with obesity indices in 591 middle-aged men and 467 middle-aged women of a cross-sectional Austrian population. Because neither polymorphic site was likely to be a functional site, we analyzed sex-specific associations of two loci haplotype combinations with obesity indices. Significant associations with BMI (P = 0.006), waist (P = 0.01) and hip circumference (P = 0.03), and total body fat (P = 0.005) and borderline significant associations with abdominal visceral and subcutaneous fat were observed in women but not men. In women, plasma triglycerides, HDL cholesterol, and glucose significantly differed by haplotype combinations, but these associations were not maintained after statistical consideration of BMI. The haplotype combination of the double variant allele with the double-wild-type allele was associated with the lowest obesity indices, whereas homozygosity for the double-variant allele was not discriminatory among haplotype combinations. These studies suggest functional differences of PPARGC1 haplotypes in human energy metabolism and support a role of PPARGC1 in obesity. PMID- 11916955 TI - Early differential defects of insulin secretion and action in 19-year-old caucasian men who had low birth weight. AB - Several studies have linked low birth weight (LBW) and type 2 diabetes. We investigated hepatic and peripheral insulin action including intracellular glucose metabolism in 40 19-year-old men (20 LBW, 20 matched control subjects), using the hyperinsulinemic-euglycemic clamp technique at two physiological insulin levels (10 and 40 mU/m(2) per min), indirect calorimetry, and [3 (3)H]glucose. Insulin secretion was examined during an oral and intravenous glucose tolerance test. Fasting p-glucose was higher in the LBW group (5.6 +/- 0.1 vs. 5.4 +/- 0.1; P < 0.05). Basal plasma glycerol concentrations were significantly lower in the LBW group. Insulin-stimulated glycolytic flux was significantly reduced, and suppression of endogenous glucose production was enhanced in the LBW group. Nevertheless, basal and insulin-stimulated rates of whole-body peripheral glucose disposal, glucose oxidation, lipid oxidation, exogenous glucose storage, and nonoxidative glucose metabolism were similar in the two groups. Insulin secretion was reduced by 30% in the LBW group, when expressed relative to insulin sensitivity (disposition index = insulin secretion x insulin action). We propose that reduced insulin-stimulated glycolysis precedes overt insulin resistance in LBW men. A lower insulin secretion may contribute to impaired glucose tolerance and ultimately lead to diabetes. PMID- 11916957 TI - Association studies of genetic variation in the WFS1 gene and type 2 diabetes in U.K. populations. AB - Mutations in the WFS1 gene cause beta-cell death, resulting in a monogenic form of diabetes known as Wolfram syndrome. The role of variation in WFS1 in type 2 diabetes susceptibility is not known. We sequenced the WFS1 gene in 29 type 2 diabetic probands and identified 12 coding variants. We used 152 parent-offspring trios to look for familial association; the R allele at residue 456 (P = 0.04) and the H allele at residue 611 (P = 0.05) as well as the R456-H611 haplotype (P = 0.032) were overtransmitted to affected offspring from heterozygous parents. In a further cohort of 327 type 2 diabetic subjects and 357 normoglycemic control subjects, the H611 allele and the R456-H611 haplotype were present in more type 2 diabetic subjects than control subjects (one-tailed P = 0.06 and P = 0.023, respectively). In a combined analysis, the H611 allele was present in 60% of all diabetes chromosomes and 55% of all control chromosomes (odds ratio [OR] 1.24 [95% CI 1.03-1.48], P = 0.02), and the R456-H611 haplotype was significantly more frequent in type 2 diabetic subjects than in control subjects (60 vs. 54%, OR 1.29 [95% CI 1.08-1.54], P = 0.0053). Our results provide the first evidence that variation in the WFS1 gene may influence susceptibility to type 2 diabetes. PMID- 11916958 TI - Molecular analysis of Berardinelli-Seip congenital lipodystrophy in Oman: evidence for multiple loci. AB - Congenital generalized lipodystrophy (CGL) is a rare disorder characterized by the absence of body fat and insulin resistance and accompanied by other features, including acanthosis nigricans, organomegaly, hyperandrogenism, and diabetes. We have examined case subjects from 11 families in Oman with CGL. All subjects were the progeny of consanguineous marriages; therefore, a homozygosity mapping strategy was used to investigate the reported loci, 11q13 and 9q34. Three subjects could be linked to 11q13, and mutations were found within the seipin gene. An additional eight subjects were linked to 9q34, but the locus was in a 9 cM interval with no known microsatellites, so further fine mapping was not possible. However, two sibships (four subjects) did not map to either locus, raising the possibility of more than two lipodystrophy loci within the Oman population. PMID- 11916959 TI - Macrophage/microglia-specific protein Iba1 enhances membrane ruffling and Rac activation via phospholipase C-gamma -dependent pathway. AB - Iba1 is a macrophage/microglia-specific calcium-binding protein that is involved in RacGTPase-dependent membrane ruffling and phagocytosis. In this study, we introduced Iba1 into Swiss 3T3 fibroblasts and demonstrated the enhancement of platelet-derived growth factor (PDGF)-induced membrane ruffling and chemotaxis. Wortmannin treatment did not completely suppressed this enhanced membrane ruffling in Iba1-expressing cells, whereas it did in Iba1-nonexpressing cells, suggesting that the enhancement is mediated through a phosphatidylinositol 3 kinase (PI3K)-independent signaling pathway. Porcine aorta endothelial cells transfected with expression constructs of Iba1 and PDGF receptor add-back mutants were used to analyze the signaling pathway responsible for the Iba1-induced enhancement of membrane ruffling. In the absence of Iba1 expression, PDGF did not induced membrane ruffling in cells expressing the Tyr-1021 receptor mutant, which is capable of activating phospholipase C-gamma (PLC-gamma) but not PI3K. In contrast, in the presence of Iba1 expression, membrane ruffling was formed in cells expressing the Tyr-1021 mutant. In addition, Rac was shown to be activated during membrane ruffling in cells expressing Iba1 and the Tyr-1021 mutant. Furthermore, dominant negative forms of PLC-gamma completely suppressed PDGF induced Iba1-dependent membrane ruffling and Rac activation. These results indicate the existence of a novel signaling pathway where PLC-gamma activates Rac in a manner dependent on Iba1. PMID- 11916960 TI - A function for phosphoinositide 3-kinase beta lipid products in coupling beta gamma to Ras activation in response to lysophosphatidic acid. AB - Although Gbetagamma is thought to mediate mitogen-activated protein kinase (MAPK) activation in response to G protein-coupled receptor stimulation, the mechanisms involved in this pathway have not been clearly defined. Phosphoinositide 3-kinase (PI3K) has been proposed as an early intermediate in this process, but its role has remained elusive. We have observed that dominant negative mutants of p110beta, but not of p110gamma, inhibited MAPK stimulation in response to lysophosphatidic acid (LPA). The role of p110beta was located upstream from Ras. To determine which of the lipid or protein kinase activities of p110beta were important for Ras activation, we produced a mutant p110beta lacking the lipid but not the protein kinase activity. This protein displayed a dominant negative activity similar to a kinase-dead mutant, indicating that p110beta lipid kinase activity was essentially involved in Ras activation. In agreement, overexpression of the lipid phosphatase PTEN was found to specifically inhibit Ras stimulation induced by LPA. In addition, we have observed that the PH domain-containing adapter protein Gab1, which is involved in p110beta activation during LPA stimulation, is also implicated in this pathway downstream of p110beta. Indeed, both membrane redistribution and phosphorylation of Gab1 were reduced in the presence of PI3K inhibitors or dominant negative p110beta. Downstream of Gab1, the tyrosine phosphatase SHP2 was found to mediate Ras activation in response to LPA and to be recruited through PI3K and Gab1, because transfection of Gab1 mutant deficient for SHP2 binding inhibited Ras activation without interfering with PI3K activation. We conclude that LPA-induced Ras activation is mediated by a p110beta/Gab1/SHP2 pathway. Moreover, we present data indicating that p110beta is effectively the target of betagamma in this pathway, suggesting that the p110beta/Gab1/SHP2 pathway provides a novel link between betagamma and Ras by integrating two early events of LPA signaling, i.e. Gbetagamma release and tyrosine kinase receptor transactivation. PMID- 11916961 TI - Astrocytes in culture produce anandamide and other acylethanolamides. AB - Anandamide (arachidonylethanolamide) is an endocannabinoid that belongs to the acylethanolamide lipid family. It is produced by neurons in a calcium-dependent manner and acts through cannabinoid CB1 receptors. Other members of the acylethanolamide lipid family are also produced by neurons and act through G protein-coupled receptors: homo-gamma-linolenylethanolamide (HEA) and docosatetraenylethanolamide (DEA) act through CB1 receptors, palmitylethanolamide (PEA) acts through CB2-like receptors, and oleylethanolamide (OEA) acts through receptors that have not yet been cloned. Although it is clear that anandamide and other acylethanolamides play a major role in neuronal signaling, whether astrocytes also produce these lipids is unknown. We developed a chemical ionization gas chromatography/mass spectrometry method that allows femtomole detection and quantification of anandamide and other acylethanolamides. Using this method, we unambiguously detected and quantified anandamide, HEA, DEA, PEA, and OEA in mouse astrocytes in culture. Stimulation of mouse astrocytes with ionomycin, a calcium ionophore, enhanced the production of anandamide, HEA, and DEA, whereas PEA and OEA levels were unchanged. Endothelin-1, a peptide known to act on astrocytes, enhanced the production of anandamide, without affecting the levels of other acylethanolamides. These results show that astrocytes produce anandamide, HEA, and DEA in a calcium-dependent manner and that anandamide biosynthesis can be selectively stimulated under physiologically relevant conditions. The relative levels of acylethanolamides in astrocytes from rat and human were different from the relative levels of acylethanolamides in mouse astrocytes, indicating that the production of these lipids differs between species. Because astrocytes are known to express CB1 receptors and inactivate endocannabinoids, our finding strongly suggests the existence of a functional endocannabinoid signaling system in these cells. PMID- 11916962 TI - Disruption of mitochondrial beta -oxidation of unsaturated fatty acids in the 3,2 trans-enoyl-CoA isomerase-deficient mouse. AB - Cellular energy metabolism is largely sustained by mitochondrial beta-oxidation of saturated and unsaturated fatty acids. To study the role of unsaturated fatty acids in cellular lipid and energy metabolism we generated a null allelic mouse, deficient in 3,2-trans-enoyl-CoA isomerase (ECI) (eci(-/-) mouse). ECI is the link in mitochondrial beta-oxidation of unsaturated and saturated fatty acids and essential for the complete degradation and for maximal energy yield. Mitochondrial beta-oxidation of unsaturated fatty acids is interrupted in eci(-/ )mice at the level of their respective 3-cis- or 3-trans-enoyl-CoA intermediates. Fasting eci(-/-) mice accumulate unsaturated fatty acyl groups in ester lipids and deposit large amounts of triglycerides in hepatocytes (steatosis). Gene expression studies revealed the induction of peroxisome proliferator-activated receptor activation in eci(-/-) mice together with peroxisomal beta- and microsomal omega-oxidation enzymes. Combined peroxisomal beta- and microsomal omega-oxidation of the 3-enoyl-CoA intermediates leads to a specific pattern of medium chain unsaturated dicarboxylic acids excreted in the urine in high concentration (dicarboxylic aciduria). The urinary dicarboxylate pattern is a reliable diagnostic marker of the ECI genetic defect. The eci(-/-) mouse might be a model of a yet undefined inborn mitochondrial beta-oxidation disorder lacking the enzyme link that channels the intermediates of unsaturated fatty acids into the beta-oxidation spiral of saturated fatty acids. PMID- 11916963 TI - Structure-based design of beta 1,4-galactosyltransferase I (beta 4Gal-T1) with equally efficient N-acetylgalactosaminyltransferase activity: point mutation broadens beta 4Gal-T1 donor specificity. AB - beta1,4-Galactosyltransferase I (Gal-T1) normally transfers Gal from UDP-Gal to GlcNAc in the presence of Mn(2+) ion. In the presence of alpha-lactalbumin (LA), the Gal acceptor specificity is altered from GlcNAc to Glc. Gal-T1 also transfers GalNAc from UDP-GalNAc to GlcNAc, but with only approximately 0.1% of Gal-T activity. To understand this low GalNAc-transferase activity, we have carried out the crystal structure analysis of the Gal-T1.LA complex with UDP-GalNAc at 2.1-A resolution. The crystal structure reveals that the UDP-GalNAc binding to Gal-T1 is similar to the binding of UDP-Gal to Gal-T1, except for an additional hydrogen bond formed between the N-acetyl group of GalNAc moiety with the Tyr-289 side chain hydroxyl group. Elimination of this additional hydrogen bond by mutating Tyr-289 residue to Leu, Ile, or Asn enhances the GalNAc-transferase activity. Although all three mutants exhibit enhanced GalNAc-transferase activity, the mutant Y289L exhibits GalNAc-transferase activity that is nearly 100% of its Gal T activity, even while completely retaining its Gal-T activity. The steady state kinetic analyses on the Leu-289 mutant indicate that the K(m) for GlcNAc has increased compared to the wild type. On the other hand, the catalytic constant (k(cat)) in the Gal-T reaction is comparable with the wild type, whereas it is 3 5-fold higher in the GalNAc-T reaction. Interestingly, in the presence of LA, these mutants also transfer GalNAc to Glc instead of to GlcNAc. The present study demonstrates that, in the Gal-T family, the Tyr-289/Phe-289 residue largely determines the sugar donor specificity. PMID- 11916965 TI - Reversible inactivation of the tumor suppressor PTEN by H2O2. AB - The tumor suppressor PTEN regulates cell migration, growth, and survival by removing the 3'-phosphate of phosphoinositides. Exposure of purified PTEN or of cells to H(2)O(2) resulted in inactivation of PTEN in a time- and H(2)O(2) concentration-dependent manner. Analysis of various cysteine mutants, including mass spectrometry of tryptic peptides, indicated that the essential Cys(124) residue in the active site of PTEN specifically forms a disulfide with Cys(71) during oxidation by H(2)O(2). The reduction of H(2)O(2)-oxidized PTEN in cells appears to be mediated predominantly by thioredoxin. Thus, thioredoxin was more efficient than glutaredoxin, glutathione, or a 14-kDa thioredoxin-like protein with regard to the reduction of oxidized PTEN in vitro. Thioredoxin co immunoprecipitated with PTEN from cell lysates; and incubation of cells with 2,4 dinitro-1-chlorobenzene (an inhibitor of thioredoxin reductase) delayed the reduction of oxidized PTEN, whereas incubation with buthioninesulfoximine (an inhibitor of glutathione biosynthesis) did not. These results suggest that the reversible inactivation of PTEN by H(2)O(2) might be important for the accumulation of 3'-phosphorylated phosphoinositides and that the uncontrolled generation of H(2)O(2) associated with certain pathological conditions might contribute to cell proliferation by inhibiting PTEN function. PMID- 11916964 TI - Reactive oxygen species differentially affect T cell receptor-signaling pathways. AB - Oxidative stress plays an important role in the induction of T lymphocyte hyporesponsiveness observed in several human pathologies including cancer, rheumatoid arthritis, leprosy, and AIDS. To investigate the molecular basis of oxidative stress-induced T cell hyporesponsiveness, we have developed an in vitro system in which T lymphocytes are rendered hyporesponsive by co-culture with oxygen radical-producing activated neutrophils. We have observed a direct correlation between the level of T cell hyporesponsiveness induced and the concentration of reactive oxygen species produced. Moreover, induction of T cell hyporesponsiveness is blocked by addition of N-acetyl cysteine, Mn(III)tetrakis(4 benzoic acid)porphyrin chloride, and catalase, confirming the critical role of oxidative stress in this system. The pattern of tyrosine-phosphorylated proteins was profoundly altered in hyporesponsive as compared with normal T cells. In hyporesponsive T cells, T cell receptor (TCR) ligation no longer induced phospholipase C-gamma1 activation and caused reduced Ca(2+) flux. In contrast, despite increased levels of ERK1/2 phosphorylation, TCR-dependent activation of mitogen-activated protein kinase ERK1/2 was unaltered in hyporesponsive T lymphocytes. A late TCR-signaling event such as caspase 3 activation was as well unaffected in hyporesponsive T lymphocytes. Our data indicate that TCR-signaling pathways are differentially affected by physiological levels of oxidative stress and would suggest that although "hyporesponsive" T cells have lost certain effector functions, they may have maintained or gained others. PMID- 11916966 TI - A novel tricomplex of BRCA1, Nmi, and c-Myc inhibits c-Myc-induced human telomerase reverse transcriptase gene (hTERT) promoter activity in breast cancer. AB - Germ-line mutations in BRCA1 predispose individuals to breast and ovarian cancers. We observed a novel endogenous association of BRCA1 with Nmi (N-Myc interacting protein) in breast cancer cells. Nmi was found to interact specifically with BRCA1, both in vitro and in vivo, by binding to two major domains in BRCA1, amino acid residues 298-683 and 1301-1863. Homodimerization of Nmi enhanced its association with BRCA1. Nmi functioned as an adaptor molecule to recruit c-Myc to a complex containing Nmi.c-Myc.BRCA1. Because c-Myc can activate transcription of the human telomerase reverse transcriptase gene (hTERT), we addressed the role of BRCA1 and Nmi in modulating c-Myc-induced hTERT promoter activity. Although Nmi or BRCA1 alone had no effect on c-Myc induced hTERT promoter activity, BRCA1 together with Nmi significantly inhibited this c-Myc induced hTERT promoter activity ( approximately 75% inhibition). Two mutated forms of BRCA1, a missense (A1708E) and a nonsense (Y1853X) that have been identified in familial breast cancers, associated with Nmi and c-Myc but failed to suppress c-Myc-induced hTERT promoter activity. These results demonstrate a novel pathogenic mechanism whereby mutations in BRCA1, via a novel transcription factor complex containing BRCA1, c-Myc, and Nmi, impair inhibition of c-Myc induced hTERT promoter activity, which allows sustained activation of telomerase, a key enzyme in carcinogenesis. PMID- 11916967 TI - Liganded androgen receptor interaction with beta-catenin: nuclear co-localization and modulation of transcriptional activity in neuronal cells. AB - A yeast two-hybrid assay was employed to identify androgen receptor (AR) protein partners in gonadotropin-releasing hormone neuronal cells. By using an AR deletion construct (AR-(Delta371-485)) as a bait, beta-catenin was identified as an AR-interacting protein from a gonadotropin-releasing hormone neuronal cell library. Immunolocalization of co-transfected AR and FLAG-beta-catenin demonstrated that FLAG-beta-catenin was predominantly cytoplasmic in the absence of androgen. In the presence of 5alpha-dihydrotestosterone, FLAG-beta-catenin completely co-localized to the nucleus with AR. This effect was specific to AR because liganded progesterone, glucocorticoid, or estrogen alpha receptors did not translocate FLAG-beta-catenin to the nucleus. Agonist-bound AR was required because the AR antagonists casodex and hydroxyflutamide failed to translocate beta-catenin. Time course experiments demonstrated that co-translocation occurred with similar kinetics. Nuclear co-localization was independent of the glycogen synthase kinase-3beta, p42/44 ERK mitogen-activated protein kinase, and phosphatidylinositol 3-kinase pathways because inhibitors of these pathways had no effect. Transcription assays demonstrated that liganded AR repressed beta catenin/T cell factor-responsive reporter gene activity. Conversely, co expression of beta-catenin/T cell factor repressed AR stimulation of AR responsive reporter gene activity. Our data suggest that liganded AR shuttles beta-catenin to the nucleus and that nuclear interaction of AR with beta-catenin may modulate transcriptional activity in androgen target tissues. PMID- 11916968 TI - The roles of ATF3 in liver dysfunction and the regulation of phosphoenolpyruvate carboxykinase gene expression. AB - Activating transcription factor 3 (ATF3), a member of the ATF/cAMP-responsive element-binding protein family of transcription factors, is a transcriptional repressor, and the expression of its corresponding gene, ATF3, is induced by many stress signals. In this report, we demonstrate that transgenic mice expressing ATF3 in the liver had symptoms of liver dysfunction such as high levels of serum bilirubin, alkaline phosphatase, alanine transaminase, aspartate transaminase, and bile acids. In addition, these mice had physiological responses consistent with hypoglycemia including a low insulin:glucagon ratio in the serum and reduced adipose tissue mass. Electrophoretic mobility shift assays indicated that ATF3 bound to the ATF/cAMP-responsvie element site derived from the promoter of the gene encoding the gluconeogenic enzyme phosphoenolpyruvate carboxykinase (PEPCK). Furthermore, transient transfection assays indicated that ATF3 repressed the activity of the PEPCK promoter. Taken together, our results are consistent with the model that the expression of ATF3 in the liver results in defects in glucose homeostasis by repressing gluconeogenesis. Because ATF3 is a stress-inducible gene, these mice may provide a model to investigate the molecular mechanisms of some stress-associated liver diseases. PMID- 11916969 TI - Human surfactant protein D (SP-D) binds Mycoplasma pneumoniae by high affinity interactions with lipids. AB - Increasing evidence now identifies surfactant protein D (SP-D) as an important element of the innate immune system of the lung. In this study, we examined the interactions of rat and human SP-D with the human pathogen, Mycoplasma pneumoniae. Rat and human SP-D bound the organism with high affinity in a reaction that required Ca(2+) and was inhibited by EGTA. Membranes derived from the organism bound the proteins in a similar manner, except the rat SP-D also exhibited a significant level of Ca(2+)-independent binding. Pretreatment of membranes with proteases did not alter the Ca(2+)-dependent SP-D binding of membranes by either protein. Mannose, glucose, maltose, and inositol, at millimolar concentrations, competed for human SP-D binding to the bacterial membrane. Lipids extracted from membranes and separated by two-dimensional thin layer chromatography bound human SP-D with high affinity in a Ca(2+)-dependent reaction. A tandem mutant of SP-D with E321Q and N323D substitutions, failed to bind M. pneumoniae lipids, directly implicating the carbohydrate recognition domain in the interaction. The interaction of rat and human SP-D with M. pneumoniae was unaffected by the presence of surfactant lipids and the hydrophobic surfactant proteins. These findings demonstrate that M. pneumoniae is likely to be recognized by SP-D in the alveolar environment and that primary determinants recognized on the organism are lipid components of the cell membrane. PMID- 11916970 TI - GNIP, a novel protein that binds and activates glycogenin, the self-glucosylating initiator of glycogen biosynthesis. AB - Glycogenin is a self-glucosylating protein involved in the initiation of glycogen biosynthesis. Self-glucosylation leads to the formation of an oligosaccharide chain, which, when long enough, supports the action of glycogen synthase to elongate it and form a mature glycogen molecule. To identify possible regulators of glycogenin, the yeast two-hybrid strategy was employed. By using rabbit skeletal muscle glycogenin as a bait, cDNAs encoding three different proteins were isolated from the human skeletal muscle cDNA library. Two of the cDNAs encoded glycogenin and glycogen synthase, respectively, proteins known to be interactors. The third cDNA encoded a polypeptide of unknown function and was designated GNIP (glycogenin interacting protein). Northern blot analysis revealed that GNIP mRNA is highly expressed in skeletal muscle. The gene for GNIP generates at least four isoforms by alternative splicing. The largest isoform GNIP1 contains, from NH(2)- to COOH-terminal, a RING finger, a B box, a putative coiled-coil region, and a B30.2-like motif. The previously identified protein TRIM7 (tripartite motif containing protein 7) is also derived from the GNIP gene and is composed of the RING finger, B box, and coiled-coil regions. The GNIP2 and GNIP3 isoforms consist of the coiled-coil region and B30.2-like domain. Physical interaction between GNIP2 and glycogenin was confirmed by co-immunoprecipitation, and in addition GNIP2 was shown to stimulate glycogenin self-glucosylation 3-4 fold. GNIPs may represent a novel participant in the initiation of glycogen synthesis. PMID- 11916971 TI - A novel intragenic sequence enhances initiator-dependent transcription in human embryonic kidney 293 cells. AB - In a variety of Drosophila TATA-less promoters, transcription is directed by initiator (Inr) sequences, which are faithfully and efficiently recognized only when flanked 3' by the downstream promoter element (DPE). This motif, which is conserved at approximately 30 bp from the RNA start site, is viewed as a downstream counterpart to the TATA box, and is recognized by the general transcription factor (TF) IID. By transient expression assays in human embryonic kidney 293 cells, we show that DE1 (distal element 1), a DNA motif located at residues +23 to +29, sustains faithful Inr-dependent transcription as efficiently as the DPE. Transcription significantly increased when DE1 and DPE sequences were adjacently placed on the same template. Results emerging from in vivo RNA analyses matched electrophoretic mobility shift assay data. In agarose electrophoretic mobility shift assays, retarded DNA-protein complexes resulting from the interaction of human holo-TFIID with either Inr(+)/DPE(+) or Inr(+)/DE1(+) promoters were formed at comparable levels, whereas binding of TFIID to both DE1 and DPE motifs was 2-fold increased. The strict requirement for spacing between the Inr and DPE was not observed for DE1, as locating the motif 4 bp away from the +1 site did not impair transcriptional enhancement. DE1 sequences may be common to many promoters and be overlooked because of their poor sequence homology. PMID- 11916972 TI - Formation of disulfide bridges by a single-chain Fv antibody in the reducing ectopic environment of the plant cytosol. AB - Disulfide bridge formation in the reducing environment of the cytosol is considered a rare event and is mostly linked to inactivation of protein activity. In this report the in vivo redox state of a single-chain Fv (scFv) antibody fragment in the plant cytosol was investigated. The scFv antibody fragment consists of the variable light and heavy chain domains from a mouse IgG antibody, which are connected by a flexible linker peptide. In each domain one disulfide bridge is present. The functionality of antibodies, which are normally secreted via the oxidizing environment of the endoplasmic reticulum, depends on the formation of intramolecular disulfide bridges. We demonstrate that a scFv can form intramolecular disulfide bridges and is functionally expressed in the cytosol of stably transformed plants. In addition, the formation of intermolecular disulfide bridges through a cysteine present in the linker peptide was observed. In contrast, transient expression in tobacco protoplasts resulted in a cytosolic scFv lacking disulfide bridges, which had a substantially reduced affinity for the antigen. This indicates that functionality rather than stability is determined by the presence of disulfide bridges in the in planta-expressed scFv antibody. The controversial observation of disulfide bond formation in the cytosol is discussed. PMID- 11916974 TI - Regulation of Hsp90 ATPase activity by the co-chaperone Cdc37p/p50cdc37. AB - In vivo activation of client proteins by Hsp90 depends on its ATPase-coupled conformational cycle and on interaction with a variety of co-chaperone proteins. For some client proteins the co-chaperone Sti1/Hop/p60 acts as a "scaffold," recruiting Hsp70 and the bound client to Hsp90 early in the cycle and suppressing ATP turnover by Hsp90 during the loading phase. Recruitment of protein kinase clients to the Hsp90 complex appears to involve a specialized co-chaperone, Cdc37p/p50(cdc37), whose binding to Hsp90 is mutually exclusive of Sti1/Hop/p60. We now show that Cdc37p/p50(cdc37), like Sti1/Hop/p60, also suppresses ATP turnover by Hsp90 supporting the idea that client protein loading to Hsp90 requires a "relaxed" ADP-bound conformation. Like Sti1/Hop/p60, Cdc37p/p50(cdc37) binds to Hsp90 as a dimer, and the suppressed ATPase activity of Hsp90 is restored when Cdc37p/p50(cdc37) is displaced by the immunophilin co-chaperone Cpr6/Cyp40. However, unlike Sti1/Hop/p60, which can displace geldanamycin upon binding to Hsp90, Cdc37p/p50(cdc37) forms a stable complex with geldanamycin bound Hsp90 and may be sequestered in geldanamycin-inhibited Hsp90 complexes in vivo. PMID- 11916973 TI - Localization of the ethylene receptor ETR1 to the endoplasmic reticulum of Arabidopsis. AB - The ethylene receptor ETR1 of Arabidopsis contains transmembrane domains responsible for ethylene binding and membrane localization. Sequence analysis does not provide information as to which membrane system of the plant cell ETR1 is localized. Examination by aqueous two-phase partitioning, sucrose density gradient centrifugation, and immunoelectron microscopy indicates that ETR1 is predominantly localized to the endoplasmic reticulum. Localization of ETR1 showed no change following a cycloheximide chase. Ethylene binding by ETR1 did not affect localization to the endoplasmic reticulum, based upon analysis of plants treated with the ethylene precursor 1-aminocyclopropane- 1-carboxylic acid and by examination of a mutant receptor that does not bind ethylene. Determinants within the amino-terminal half of ETR1 are sufficient for targeting to and retention at the endoplasmic reticulum. These data support a central role of the plant endoplasmic reticulum in hormone perception and signal transduction. PMID- 11916975 TI - The antiviral dynamin family member, MxA, tubulates lipids and localizes to the smooth endoplasmic reticulum. AB - Mx proteins are induced by type I interferon and inhibit a broad range of viruses by undefined mechanisms. They are included within the dynamin family of large GTPases, which are involved in vesicle trafficking and share common biophysical features. These properties include the propensity to self-assemble, an affinity for lipids, and the ability to tubulate membranes. In this report we establish that human MxA, despite sharing only 30% homology with conventional dynamin, possesses many of these properties. We demonstrate for the first time that MxA self-assembles into rings that tubulate lipids in vitro, and associates with a specific membrane compartment in cells, the smooth endoplasmic reticulum. PMID- 11916976 TI - Lateral dimerization of the E-cadherin extracellular domain is necessary but not sufficient for adhesive activity. AB - Cadherins are transmembrane glycoproteins involved in Ca(2+)-dependent cell-cell adhesion. Using L cells coexpressing E-cadherin constructs with different epitope tags, we examined the lateral dimerization of E-cadherin and its adhesive activity by co-immunoprecipitation and aggregation assays, respectively. Although the transmembrane domain is required for dimerization, tail-less constructs possessing the transmembrane domain of either N-cadherin or CD45 show dimerization and are active in aggregation assays. Two mutant constructs having either of two amino acid substitutions, W2A or substitutions that disrupt the recognition sequence for endoproteolytic enzymes involved in removal of the precursor segment, cannot form dimers and are inactive in aggregation. These monomeric proteins, like their wild-type dimerizing counterparts, retain their Ca(2+)-dependent resistance to trypsin digestion, suggesting that dimerization per se does not induce a large conformational change. Two other constructs, having either an amino acid substitution, D134A, or a C-terminal deletion of 70 amino acid residues, retain the ability to associate laterally but are inactive in aggregation assays. Staurosporine treatment of cells expressing the latter construct increases aggregation but does not increase the extent of lateral dimerization. Thus, lateral dimerization is necessary, but not sufficient for adhesive activity. PMID- 11916977 TI - Multiple regulatory roles of a novel Saccharomyces cerevisiae protein, encoded by YOL002c, in lipid and phosphate metabolism. AB - The yeast open reading frame YOL002c encodes a putative membrane protein. This protein is evolutionarily conserved across species, including humans, although the function of each of these proteins remains unknown. YOL002c is highly expressed in yeast cells that are grown in the presence of saturated fatty acids such as myristate. Furthermore, cells in which the YOL002c gene is disrupted grow poorly on this carbon source. These mutant cells are also resistant to the polyene antibiotic, nystatin. Gene chip analysis on yol002cDelta cells revealed that a variety of genes encoding proteins involved in fatty acid metabolism and in the phosphate signaling pathway are induced in this mutant strain. In addition, our studies demonstrated that in the disruption strain acid phosphatase activity is expressed constitutively, and the cells accumulate polyphosphate to much higher levels than wild-type cells. A homologous human protein is able to partially rescue these defects in phosphate metabolism. We propose that YOL002c encodes a Saccharomyces cerevisiae protein that plays a key role in metabolic pathways that regulate lipid and phosphate metabolism. PMID- 11916978 TI - Protein kinase C (PKC) delta regulates PKCalpha activity in a Syndecan-4 dependent manner. AB - The phosphorylation state of Ser(183) in the cytoplasmic tail of syndecan-4 determines the binding affinity of the cytoplasmic tail to phosphatidylinositol 4,5-bisphosphate (PIP(2)), the capacity of the tail to multimerize, and its ability to activate protein kinase C (PKC) alpha. We sought to identify the kinase responsible for this phosphorylation and to determine its downstream effects on PKCalpha activity and on endothelial cell function. Among several PKC isoenzymes tested, only PKCalpha and -delta were able to specifically phosphorylate Ser(183) in vitro. However, studies in cultured endothelial cells showed that the phosphorylation level of syndecan-4 was significantly reduced in endothelial cells expressing a dominant negative (DN) PKCdelta but not a DN PKCalpha mutant. Syndecan-4/PIP(2)-dependent PKCalpha activity was significantly increased in PKCdelta DN cells, while PKCdelta overexpression was accompanied by decreased PKCalpha activity. PKCdelta-overexpressing cells exhibited a significantly lower proliferation rate and an impaired tube formation in response to FGF2, which were mirrored by similar observations in PKCalpha DN endothelial cells. These findings suggest that PKCdelta is the kinase responsible for syndecan-4 phosphorylation, which, in turn, attenuates the cellular response to FGF2 by reducing PKCalpha activity. The reduced PKCalpha activity then leads to impaired endothelial cell function. We conclude that PKCdelta regulates PKCalpha activity in a syndecan-4-dependent manner. PMID- 11916979 TI - The intraflagellar transport protein, IFT88, is essential for vertebrate photoreceptor assembly and maintenance. AB - Approximately 10% of the photoreceptor outer segment (OS) is turned over each day, requiring large amounts of lipid and protein to be moved from the inner segment to the OS. Defects in intraphotoreceptor transport can lead to retinal degeneration and blindness. The transport mechanisms are unknown, but because the OS is a modified cilium, intraflagellar transport (IFT) is a candidate mechanism. IFT involves movement of large protein complexes along ciliary microtubules and is required for assembly and maintenance of cilia. We show that IFT particle proteins are localized to photoreceptor connecting cilia. We further find that mice with a mutation in the IFT particle protein gene, Tg737/IFT88, have abnormal OS development and retinal degeneration. Thus, IFT is important for assembly and maintenance of the vertebrate OS. PMID- 11916980 TI - Bloom's syndrome protein is required for correct relocalization of RAD50/MRE11/NBS1 complex after replication fork arrest. AB - Bloom's syndrome (BS) is a rare genetic disorder characterized by a broad range of symptoms and, most importantly, a predisposition to many types of cancers. Cells derived from patients with BS exhibit an elevated rate of somatic recombination and hypermutability, supporting a role for bleomycin (BLM) in the maintenance of genomic integrity. BLM is thought to participate in several DNA transactions, the failure of which could give raise to genomic instability, and to interact with many proteins involved in replication, recombination, and repair. In this study, we show that BLM function is specifically required to properly relocalize the RAD50/MRE11/NBS1 (RMN) complex at sites of replication arrest, but is not essential in the activation of BRCA1 either after stalled replication forks or gamma-rays. We also provide evidence that BLM is phosphorylated after replication arrest in an Ataxia and RAD3-related protein (ATR)-dependent manner and that phosphorylation is not required for subnuclear relocalization. Therefore, in ATR dominant negative mutant cells, the assembly of the RMN complex in nuclear foci after replication blockage is almost completely abolished. Together, these results suggest a relationship between BLM, ATR, and the RMN complex in the response to replication arrest, proposing a role for BLM protein and RMN complex in the resolution of stalled replication forks. PMID- 11916981 TI - Mammalian class E vps proteins recognize ubiquitin and act in the removal of endosomal protein-ubiquitin conjugates. AB - There is increasing evidence that ubiquitination of receptors provides an important endosomal sorting signal. Here we report that mammalian class E vacuolar protein-sorting (vps) proteins recognize ubiquitin. Both tumor susceptibility gene 101 (TSG101)/human VPS (hVPS)28 and hepatocyte growth factor receptor substrate (Hrs) cytosolic complexes bind ubiquitin-agarose. TSG101 and hVPS28 are localized to endosomes that contain internalized EGF receptor and label strongly for ubiquitinated proteins. Microinjection of anti-hVPS28 specifically retards EGF degradation and leads to endosomal accumulation of ubiquitin-protein conjugates. Likewise, depletion of TSG101 impairs EGF trafficking and causes dramatic relocalization of ubiquitin to endocytic compartments. Similar defects are found in cells overexpressing Hrs, further emphasizing the links between class E protein function, receptor trafficking, and endosomal ubiquitination. PMID- 11916984 TI - Is transdifferentiation in trouble? AB - Spectacular examples of transdifferentiation--such as brain cells turning to blood and blood to brain--have given way to sneaking suspicions about artifacts in culture, fusion, and clonality. Could cell fates be relatively fixed after all? PMID- 11916982 TI - A cycle of Vam7p release from and PtdIns 3-P-dependent rebinding to the yeast vacuole is required for homotypic vacuole fusion. AB - Vacuole fusion requires a coordinated cascade of priming, docking, and fusion. SNARE proteins have been implicated in the fusion itself, although their precise role in the cascade remains unclear. We now report that the vacuolar SNAP-23 homologue Vam7p is a mobile element of the SNARE complex, which moves from an initial association with the cis-SNARE complex via a soluble intermediate to the docking site. Soluble Vam7p is specifically recruited to vacuoles and can rescue a fusion reaction poisoned with antibodies to Vam7p. Both the recombinant Vam7p PX domain and a FYVE domain construct of human Hrs block the recruitment of Vam7p and vacuole fusion, demonstrating that phosphatidylinositol 3-phosphate is a primary receptor of Vam7p on vacuoles. We propose that the Vam7p cycle is linked to the availability of a lipid domain on yeast vacuoles, which is essential for coordinating the fusion reaction prior to and beyond docking. PMID- 11916985 TI - Comparative neuroethology of feeding control in molluscs. AB - Over the last 30 years, many laboratories have examined, in parallel, the feeding behaviour of gastropod molluscs and the properties of the nervous system that give rise to this behaviour. Equal attention to both behavioural and neurobiological issues has provided deep insight into the functioning of the nervous system in generating and controlling behaviour. The conclusions derived from studies on gastropod feeding are generally consistent with those from other systems, but often provide more detailed information on the behavioural function of a particular property of the nervous system. A review of the literature on gastropod feeding illustrates a number of important messages. (i) Many of the herbivorous gastropods display similarities in behaviour that are reflected in corresponding similarities in neural anatomy, pharmacology and physiology. By contrast, the same aspects of the behaviour of different carnivorous species are quite variable, possibly because of their specialised prey-capture techniques. Nonetheless, some aspects of the neural control of feeding are preserved. (ii) Feeding in all species is flexible, with the behaviour and the physiology adapting to changes in the current environment and internal state and as a result of past experience. Flexibility arises via processes that may take place at many neural sites, and much of the modulation underlying behavioural flexibility is understood at a systems and at a cellular level. (iii) Neurones seem to have specific functions that are consistent with their endogenous properties and their synaptic connections, suggesting that individual neurones code specific pieces of information (i.e. they are 'grandmother cells'). However, the properties of a neurone can be extremely complex and can be understood only in the context of the complete neural circuit and the behaviour that it controls. In systems that are orders of magnitude more complex, it would be impossible to understand the functional properties of an individual neurone, even if it also coded specific information. (iv) Systems such as gastropod feeding may provide a model for understanding the functional properties of more complex systems. PMID- 11916983 TI - Analysis of oxysterol binding protein homologue Kes1p function in regulation of Sec14p-dependent protein transport from the yeast Golgi complex. AB - Oxysterol binding proteins (OSBPs) comprise a large conserved family of proteins in eukaryotes. Their ubiquity notwithstanding, the functional activities of these proteins remain unknown. Kes1p, one of seven members of the yeast OSBP family, negatively regulates Golgi complex secretory functions that are dependent on the action of the major yeast phosphatidylinositol/phosphatidylcholine Sec14p. We now demonstrate that Kes1p is a peripheral membrane protein of the yeast Golgi complex, that localization to the Golgi complex is required for Kes1p function in vivo, and that targeting of Kes1p to the Golgi complex requires binding to a phosphoinositide pool generated via the action of the Pik1p, but not the Stt4p, PtdIns 4-kinase. Localization of Kes1p to yeast Golgi region also requires function of a conserved motif found in all members of the OSBP family. Finally, we present evidence to suggest that Kes1p may regulate adenosine diphosphate ribosylation factor (ARF) function in yeast, and that it may be through altered regulation of ARF that Kes1p interfaces with Sec14p in controlling Golgi region secretory function. PMID- 11916986 TI - K(+) transport in the mesonephric collecting duct system of the toad Bufo bufo: microelectrode recordings from isolated and perfused tubules. AB - We studied the mechanisms of K(+) transport in cells from isolated and perfused collecting tubules and ducts from the mesonephric kidney of the toad Bufo bufo. Cells were impaled with microelectrodes across the basal cell membrane. The basolateral membrane potential (V(bl)) depolarized upon change of bath [K(+)] from 3 to 20 mmol l(-1) demonstrating a large K(+) conductance in this membrane. In collecting tubules and collecting ducts a V(bl) of -66+/-2 mV and -74+/-4 mV depolarized by 30+/-2 mV and 36+/-3 mV, respectively (N=23; 15). The K(+) channel inhibitor Ba(2+) (1 mmol l(-1)) inhibited the basolateral K(+) conductance and depolarized a V(bl) of -64+/-4 mV by 30+/-6 mV (N=8). Luminal K(+) steps (3 to 20 mmol l(-1)) demonstrated a K(+) conductance in the apical cell membrane. In collecting tubules and collecting ducts a V(bl) of -70+/-3 mV and -73+/-3 mV depolarized by 11+/-3 mV and 16+/-3 mV, respectively (N=11; 11). This conductance could also be inhibited by Ba(2+), which depolarized a V(bl) of -71+/-5 mV by 9+/ 3 mV (N=5). The pump inhibitor ouabain (1 mmol l(-1)) depolarized V(bl), but addition of furosemide to bath solution did not affect V(bl). The [K(+)] in urine varied from 1.3 to 22.8 mmol l(-1). In conclusion, we propose that the collecting duct system of B. bufo secretes K(+) into the urine via luminal K(+) channels. PMID- 11916987 TI - Harmonic oscillatory orientation relative to the wind in nocturnal roosting flights of the swift Apus apus. AB - Swifts regularly spend the night flying at high altitude. From previous studies based on tracking radar observations, we know that they stay airborne during the night and prefer to orient themselves into the wind direction with an increased angular concentration with increasing wind speed. In this study, we investigated the orientation relative to the wind of individual swifts by frequency (discrete Fourier transform) and autocorrelation analysis based on time series (10s intervals) of the angle between the swifts' heading and the wind direction for radar trackings of long duration (9-60 min). The swifts often showed a significant harmonic oscillation of their heading direction relative to the wind, with a frequency mostly in the range 1-17 mHz, corresponding to cycle periods of 1-16 min. The swifts also sometimes performed circling flights at low wind speeds. Wind speed ranged from 1.3 to 14.8 m s(-1), and we expected to find different patterns of orientation at different wind speeds, assuming that the swifts adapt their orientation to avoid substantial displacement during their nocturnal flights. However, oscillatory orientation was found at all wind speeds with variable frequencies/periods that did not show any consistent relationship with wind speed. It remains to be shown whether cyclic heading changes are a regular feature of bird orientation. PMID- 11916988 TI - In vivo membrane trafficking role for an insect N-ethylmaleimide-sensitive factor which is developmentally regulated in endocrine cells. AB - The hexameric ATPase, N-ethylmaleimide-sensitive factor (NSF) is implicated in the release of neurotransmitters and in mediating fusion between intracellular membranes. Due to the conservation of proteins in constitutive and regulated membrane fusion reactions, NSF and its downstream targets have been predicted also to participate in fusion reactions underlying endocrine function, but there is little experimental evidence to support such a role for NSF in insect neuroendocrine secretion. Here we have characterized the NSF orthologue (MsNSF) from the endocrine model for development Manduca sexta. MsNSF is developmentally regulated in endocrine organs of the protocerebral complex. Enrichment of MsNSF in corpora cardiaca (CC) and not in corpora allata (CA) indicates that it might play a preferential role in releasing hormones produced in CC. Endocrine/paracrine cells of the enteric system in M. sexta exhibit selective MsNSF enrichment. Together the data point to a more selective participation of MsNSF in development of M. sexta by its involvement in a subset of factors, whereas other as-yet-unidentified homolog(s) might regulate secretion from CA and a large set of endocrine/paracrine cells. We further characterized the in vivo role of MsNSF by heterologous expression. In contrast to vertebrate NSF, MsNSF is functional in yeast membrane fusion in vivo. MsNSF rectifies defects in SEC18 (yeast NSF homologue) at nearly all discernible steps where Sec18p has been implicated in the biosynthetic route. This underscores the utility of our approach to delineate functional roles for proteins from systems that are not currently amenable to in vitro reconstitution. PMID- 11916989 TI - Visual pigments and oil droplets in diurnal lizards: a comparative study of Caribbean anoles. AB - We report microspectrophotometric (MSP) data for the visual pigments and oil droplets of 17 species of Caribbean anoline lizard known to live in differing photic habitats and having distinctly different dewlap colors. The outgroup Polychrus marmoratus was also examined to gain insight into the ancestral condition. Except for Anolis carolinensis, which is known to use vitamin A(2) as its visual pigment chromophore, all anoline species examined possessed at least four vitamin-A(1)-based visual pigments with maximum absorbance (lambda(max)) at 564, 495, 455 and 365 nm. To the previously reported visual pigments for A. carolinensis we add an ultraviolet-sensitive one with lambda(max) at 365 nm. Five common classes of oil droplet were measured, named according to apparent color and associated with specific cone classes - yellow and green in long-wavelength sensitive (LWS) cones, green only in medium-wavelength-sensitive (MWS) cones and colorless in short-wavelength-sensitive (SWS) and ultraviolet-sensitive (UVS) cones. MSP data showed that the colorless droplet in the SWS cone had significant absorption between 350 and 400 nm, while the colorless droplet in the UVS cone did not. The pattern for Polychrus marmoratus was identical to that for the anoles except for the presence of a previously undescribed visual cell with a rod like outer segment, a visual pigment with a lambda(max) of 497 nm and a colorless oil droplet like that in the UVS cones. These findings suggest that anoline visual pigments, as far as they determine visual system spectral sensitivity, are not necessarily adapted to the photic environment or to the color of significant visual targets (e.g. dewlaps). PMID- 11916990 TI - Kinematics of the buccal mass during swallowing based on magnetic resonance imaging in intact, behaving Aplysia californica. AB - A novel magnetic resonance imaging interface has been developed that makes it possible to image movements in intact, freely moving subjects. We have used this interface to image the internal structures of the feeding apparatus (i.e. the buccal mass) of the marine mollusc Aplysia californica. The temporal and spatial resolution of the resulting images is sufficient to describe the kinematics of specific muscles of the buccal mass and the internal movements of the main structures responsible for grasping food, the radula and the odontophore. These observations suggest that a previously undescribed feature on the anterior margin of the odontophore, a fluid-filled structure that we term the prow, may aid in opening the jaw lumen early in protraction. Radular closing during swallowing occurs near the peak of protraction as the radular stalk is pushed rapidly out of the odontophore. Retraction of the odontophore is enhanced by the closure of the lumen of the jaws on the elongated odontophore, causing the odontophore to rotate rapidly towards the esophagus. Radular opening occurs after the peak of retraction and without the active contraction of the protractor muscle 12 and is due, in part, to the movement of the radular stalk into the odontophore. The large variability between responses also suggests that the great flexibility of swallowing responses may be due to variability in neural control and in the biomechanics of the ingested food and to the inherent flexibility of the buccal mass. PMID- 11916991 TI - A model of scale effects in mammalian quadrupedal running. AB - Although the effects of body size on mammalian locomotion are well documented, the underlying mechanisms are not fully understood. Here, we present a computational model of the mechanics, control and energetics that unifies some well-known scale effects in running quadrupeds. The model consists of dynamic, physics-based simulations of six running mammals ranging in size from a chipmunk to a horse (0.115-676 kg). The 'virtual animals' are made up of rigid segments (head, trunk and four legs) linked by joints and are similar in morphology to particular species. In the model, each stance limb acts as a spring operating within a narrow range of stiffness, forward motion is powered and controlled by active hip and shoulder torques, and metabolic cost is predicted from the time course of supporting body weight. Model parameters that are important for stability (joint stiffnesses, limb-retraction times and target positions and velocities of the limbs) are selected such that (i) running kinematics (aerial height, forward speed and body pitch) is smooth and periodic and (ii) overall leg stiffness is in agreement with published data. Both trotting and galloping gaits are modeled, and comparisons across size are made at speeds that are physiologically similar among species. Model predictions are in agreement with data on vertical stiffness, limb angles, metabolic cost of transport, stride frequency, peak force and duty factor. This work supports the idea that a single, integrative model can predict important features of running across size by employing simple strategies to control overall leg stiffness. More broadly, the model provides a quantitative framework for testing hypotheses that relate limb control, stability and metabolic cost. PMID- 11916992 TI - Effects of temperature on sustained swimming performance and swimming kinematics of the chub mackerel Scomber japonicus. AB - The effects of a 6 degrees C difference in water temperature on maximum sustained swimming speed, swimming energetics and swimming kinematics were measured in the chub mackerel Scomber japonicus (Teleostei: Scombridae), a primarily coastal, pelagic predator that inhabits subtropical and temperate transition waters of the Atlantic, Pacific and Indian Oceans. New data for chub mackerel acclimated to 18 degrees C are compared with published data from our laboratory at 24 degrees C. Twelve individuals acclimated to each of two temperatures (15.6-26.3 cm fork length, FL, and 34-179 g at 18 degrees C; 14.0-24.7 cm FL and 26-156 g at 24 degrees C) swam at a range of speeds in a temperature-controlled Brett-type respirometer, at the respective acclimation temperature. At a given fish size, the maximum speed that S. japonicus was able to maintain for a 30-min period, while swimming steadily using slow, oxidative locomotor muscle (U(max,c)), was significantly greater at 24 than at 18 degrees C (52.5-97.5 cm s(-1) at 18 degrees C and 70-120 cm s(-1) at 24 degrees C). At a given speed and fish size, the rate of oxygen consumption (VO(2)) was significantly higher at 24 than at 18 degrees C because of a higher net cost of transport (1073-4617 J km(-1) kg(-1) at 18 degrees C and 2708-14895 J km(-1) kg(-1) at 24 degrees C). Standard metabolic rate, calculated by extrapolating the logO(2) versus swimming speed relationship to zero speed, did not vary significantly with temperature or fish mass (126.4+/ 67.2 mg O(2) h(-1) kg(-1) at 18 degrees C and 143.2+/-80.3 mg O(2) h(-1) kg(-1) at 24 degrees C; means +/- S.D., N=12). Swimming kinematics was quantified from high-speed (120 Hz) video recordings analyzed with a computerized, two dimensional motion-analysis system. At a given speed and fish size, there were no significant effects of temperature on tail-beat frequency, tail-beat amplitude or stride length, but propulsive wavelength increased significantly with temperature as a result of an increase in propulsive wave velocity. Thus, the main effects of temperature on chub mackerel swimming were increases in both U(max,c) and the net cost of swimming at 24 degrees C. Like other fishes, S. japonicus apparently must recruit more slow, oxidative muscle fibers to swim at a given sustainable speed at the lower temperature because of the reduced power output. Thus, the 24 degrees C mackerel reach a higher speed before they must recruit the fast, glycolytic fibers, thereby increasing U(max,c) at 24 degrees C. By quantifying in vivo the effects of temperature on the swimming performance of an ectothermic species that is closely related to the endothermic tunas, this study also provides evidence that maintaining the temperature of the slow, oxidative locomotor muscle at 6 degrees C or more above ambient water temperature in tunas should significantly increase sustainable swimming speeds, but also increase the energetic cost of swimming, unless cardiac output limits muscle performance. PMID- 11916993 TI - Effect of muscle temperature on rate of oxygen uptake during exercise in humans at different contraction frequencies. AB - The effect of elevated human muscle temperature on energy turnover was investigated during cycling exercise (at 85 % of (VO(2)max)) at a contraction frequency of 60 revs min(-1). Muscle temperature was passively elevated prior to exercise by immersion of the legs in a hot water bath (42 degrees C). During exercise at this low pedalling rate, total energy turnover was higher (P<0.05) when muscle temperature was elevated compared with normal temperature (70.4+/-3.7 versus 66.9+/-2.4 kJ min(-1), respectively). Estimated net mechanical efficiency was found to be lower when muscle temperature was elevated. A second experiment was conducted in which the effect of elevated human muscle temperature on energy turnover was investigated during cycling exercise (at 85 % of (VO(2)max)) at a contraction frequency of 120 revs min(-1). Under the conditions of a high pedalling frequency, an elevated muscle temperature resulted in a lower energy turnover (P<0.05) compared with the normal muscle temperature (64.9+/-3.7 versus 69.0+/-4.7 kJ min(-1), respectively). The estimated net mechanical efficiency was therefore higher when muscle temperature was elevated. We propose that, in these experiments, prior heating results in an inappropriately fast rate of cross bridge cycling when exercising at 60 revs min(-1), leading to an increased energy turnover and decreased efficiency. However, at the faster pedalling rate, the effect of heating the muscle shifts the efficiency/velocity relationship to the right so that cross-bridge detachment is more appropriately matched to the contraction velocity and, hence, energy turnover is reduced. PMID- 11916994 TI - Specialized olfactory receptor neurons mediating intra- and interspecific chemical communication in leafminer moths Eriocrania spp. (Lepidoptera: Eriocraniidae). AB - We performed a physiological and morphological characterization of sensilla auricillica in male Eriocrania semipurpurella moths. Each auricillic sensillum contained three olfactory receptor neurons. Responding neurons (87 of 139) could be grouped into five physiological types. Type 1 responded to (R,Z)-6-nonen-2-ol and type 2 to its enantiomer (S,Z)-6-nonen-2-ol, both of which are pheromone components of E. semipurpurella. Type 3 responded to both (R)-heptan-2-ol and (R,Z)-4-hepten-2-ol, which are pheromone components of the sympatric species E. cicatricella. Types 4 and 5 responded to the ketones (Z)-6-nonen-2-one and/or nonan-2-one, which are found in the pheromone glands of female E. semipurpurella. Field-trapping showed that type 3 receptor neurons mediate strongly antagonistic effects of (R)-heptan-2-ol and (R,Z)-4-hepten-2-ol on E. semipurpurella, while nonan-2-one should possibly be included as a synergist in the sex pheromone blend of this species. The attraction of E. cicatricella and E. sparrmannella to compounds mixed with the pheromone blend of E. semipurpurella shows that the pheromone components of E. semipurpurella have little or no antagonistic effects on these species. The morphology and physiology of eriocraniid pheromone sensilla are very similar to those found in the order Trichoptera (caddisflies), suggesting a homology between pheromone detection systems in the two sister orders Lepidoptera and Trichoptera. PMID- 11916995 TI - Modeling an electrosensory landscape: behavioral and morphological optimization in elasmobranch prey capture. AB - Most biological sensory systems benefit from multiple sensors. Elasmobranchs (sharks, skates and rays) possess an array of electroreceptive organs that facilitate prey location, mate location and navigation. Here, the perceived electrosensory landscape for an elasmobranch approaching prey is mathematically modeled. The voltages that develop simultaneously in dozens of separate sensing organs are calculated using electrodynamics. These voltages lead directly to firing rate modifications in the primary afferent nerves. The canals connecting the sense organs to an elasmobranch's surface exhibit great variation of location and orientation. Here, the voltages arising in the sense organs are found to depend strongly on the geometrical distribution of the corresponding canals. Two applications for the modeling technique are explored: an analysis of observed elasmobranch prey-capture behavior and an analysis of morphological optimization. For the former, results in specific predator-prey scenarios are compared with behavioral observations, supporting the approach algorithm suggested by A. Kalmijn. For the latter, electrosensory performance is contrasted for two geometrical models of multiple sense organs, a rounded head and a hammer-shaped head. PMID- 11916996 TI - Learning speed and contextual isolation in bumblebees. AB - Bumblebees will learn to approach one of a pair of patterns (a 45 degrees grating) and to avoid the other (a 135 degrees grating) to reach a feeder, and to do the opposite to reach their nest (approach a 135 degrees grating and avoid a 45 degrees grating). These two potentially competing visuo-motor associations are insulated from each other because they are set in different contexts. We investigated what training conditions allow the two sets of associations to be acquired without mutual interference. If the discrimination at the feeder has already been learnt, then the discrimination at the nest can be readily acquired without disrupting the bees' performance at the feeder. But, if the two are learnt simultaneously, there is mutual interference. Prior experience of the two contexts before the discriminations are learnt does not prevent interference. We conclude that visual patterns and contextual cues must already be associated with each other for a visuo-motor association to be isolated from the interfering effects of a competing association that is acquired in a separate context. This pattern of results was mimicked in a simple neural network with Hebbian synapses, in which local and contextual cues were bound together into a configural unit. PMID- 11916997 TI - Discontinuous gas-exchange in centipedes and its convergent evolution in tracheated arthropods. AB - We have examined the gas-exchange characteristics of five southern African centipede species from three orders. Two scolopendromorph species exhibit discontinuous gas-exchange cycles (DGCs) identical to those recorded for several insect and chelicerate species. Another scolopendromorph and a lithobiomorph species exhibit weak periodic patterns, and a scutigermorph species shows continuous gas exchange. A crucial component for DGCs in tracheated arthropods is the presence of occludible spiracles. However, on the basis of studies of temperate centipedes, most recent invertebrate biology texts hold the view that centipedes, as a group, cannot close their spiracles. Using flow-through normoxic and normoxic-anoxic-normoxic respirometry and electron microscopy, we conclusively demonstrate that at least one of the scolopendromorph species, Cormocephalus morsitans L., can close its spiracles fully, thus accounting for its DGCs. Homologies in spiracular structure and DGCs suggest that several other tracheated arthropod taxa probably have this ability too and that DGCs have evolved convergently at least four times in the Arthropoda. Spiracular closure and discontinuous gas-exchange cycles are probably more widespread in arthropods than has previously been suspected. PMID- 11916998 TI - Relationship between individual variation in morphological characters and swimming costs in brook charr (Salvelinus fontinalis) and yellow perch (Perca flavescens). AB - The objective of this study was to examine if individual variation in morphological characters is related to swimming costs in wild and domestic brook charr, and in wild yellow perch. Our results indicate that absolute swimming cost was higher in wild and domestic brook charr individuals having a stout body shape, and these individuals are therefore less efficient swimmers. These results are consistent with field observations that described relationships between individual variation in morphology and habitat use in salmonids. Further analyses indicated that standard metabolic rates were higher in individuals having a stout body shape, and that net swimming cost was not related to body shape. Accordingly, the higher swimming cost of stout individuals is probably an indirect consequence of an increase in standard metabolic rate. In wild yellow perch, absolute and net swimming costs were higher in individuals having a stout body shape and a low aspect caudal fin, and standard metabolic rate was not related to body shape. Therefore, in contrast to brook charr, individual variation in the swimming cost of yellow perch appears to be related to morphological characters that affect drag and thrust forces, which is consistent with previously published inter-specific observations. PMID- 11916999 TI - Variation in speed, gait characteristics and microhabitat use in lacertid lizards. AB - We quantified four gait characteristics (stride length, stride frequency, step length and floating distance) over a range of running speeds in 11 lacertid lizard species known to vary in maximal sprint speed and microhabitat use. For each species, we measured snout-vent length (SVL), body mass and hindlimb length. We tested which variables determine sprint speed, how each species modulates sprint speed and whether species occurring in different microhabitats differ in speed modulation strategy. In doing so, we aimed to test the assumption that sprint speed is correlated with hindlimb length through its effect on stride length. Variation in maximal sprint speed is determined by variation in both stride length and frequency, but those species that modulate their sprint speed mainly by altering stride length appear to attain the highest maximal speeds. At maximal sprint speed, long-limbed species take larger strides because of the positive effect of hindlimb length on step length and floating distance. However, when hindlimb length is statistically controlled for, mass has a negative effect on step length. None of the three morphological variables explained the interspecific variation in stride frequency at maximal sprint speed. Possibly, differences in physiological properties (e.g. muscle contraction speed) underlie the variation in stride frequency. The 11 species modulate their speed in different ways. Lacertids often seen in vertical microhabitats do not seem to be either pronounced stride length or frequency modulators. These species alter their speed by combining the two strategies. However, species occurring mostly in open and vegetated microhabitats are, respectively, stride length and stride frequency modulators. This difference in running style is substantiated by interspecific morphological differences. PMID- 11917000 TI - Neurodegenerative and neuroprotective effects of tumor Necrosis factor (TNF) in retinal ischemia: opposite roles of TNF receptor 1 and TNF receptor 2. AB - Tumor necrosis factor (TNF) is an important factor in various acute and chronic neurodegenerative disorders. In retinal ischemia, we show early, transient upregulation of TNF, TNF receptor 1 (TNF-R1), and TNF-R2 6 hr after reperfusion preceding neuronal cell loss. To assess the specific role of TNF and its receptors, we compared ischemia-reperfusion-induced retinal damage in mice deficient for TNF-R1, TNF-R2, or TNF by quantifying neuronal cell loss 8 d after the insult. Surprisingly, TNF deficiency did not affect overall cell loss, yet absence of TNF-R1 led to a strong reduction of neurodegeneration and lack of TNF R2 led to an enhancement of neurodegeneration, indicative of TNF-independent and TNF-dependent processes in the retina, with TNF-R1 augmenting neuronal death and TNF-R2 promoting neuroprotection. Western blot analyses of retinas revealed that reduction of neuronal cell loss in TNF-R1/ animals correlated with the presence of activated Akt/protein kinase B (PKB). Inhibition of the phosphatidylinositol 3 kinase signaling pathway reverted neuroprotection in TNF-R1-deficient mice, indicating an instrumental role of Akt/PKB in neuroprotection and TNF-R2 dependence of this pathway. Selective inhibition of TNF-R1 function may represent a new approach to reduce ischemia-induced neuronal damage, being potentially superior to strategies aimed at suppression of TNF activity in general. PMID- 11917001 TI - Alterations in the thickness of motor cortical subregions after motor-skill learning and exercise. AB - Behavioral manipulations such as housing in an enriched environment have been shown to increase brain weight and visual cortical thickness. The present study was designed to test whether skill learning or repetitive movements can alter the thickness of the motor cortex. One group of 6-mo-old Long-Evans female rats learned motor skills on an obstacle course that increased in difficulty over training and required balance and coordination. A second group ran voluntarily in exercise wheels attached to their home cage but had little opportunity for skill learning. The third group was handled daily but received no opportunity for learning or exercise. Each condition lasted 26-29 d. The skill-learning and exercise conditions had greater heart weight, and the exercise condition had greater adrenal gland weights than controls. The thickness of the motor cortex was measured in four coronal planes between -2.33 mm to -0.3 mm from bregma. Regions of interest that corresponded to published maps of forelimb and hind-limb representations were analyzed together. Rats in the skill-learning condition had significantly thicker medial cortical areas in the two anterior planes (-0.8 and 0.3 mm from bregma). These regions correspond to previously mapped hind-limb representations. The exercise group had greater thickness of the medial region at -0.8 mm from bregma. Cortical thickness in all conditions varied significantly along the medial to lateral axis. For both treatments, the effects were restricted to medial and anterior regions of interest rather than posterior or lateral regions of interest. The results indicate that robust exercise, in addition to skill learning, is capable of altering the thickness of the motor cortex, but that the effects are restricted rather than distributed within the regions studied. PMID- 11917002 TI - Mediodorsal thalamic lesions impair trace eyeblink conditioning in the rabbit. AB - Rabbits received lesions of the mediodorsal nucleus of the thalamus (MDN) or sham lesions and were subjected to classical eyeblink (EB) and heart rate (HR) conditioning. All animals received trace conditioning, with a.5-sec tone conditioned stimulus, a .5-sec trace period, and a 50-msec periorbital shock unconditioned stimulus. Animals with MDN lesions acquired the EB conditioned response (CR) more slowly than sham-lesioned animals. However, previous studies have shown that MDN damage does not affect delay conditioning using either .5-sec or 1-sec interstimulus intervals. The lesions had no significant effect on the HR CR. These results suggest that information processed by MDN and relayed to the prefrontal cortex is required for somatomotor response selection under nonoptimal learning conditions. PMID- 11917003 TI - The effects of dopamine D(1) receptor blockade in the prelimbic-infralimbic areas on behavioral flexibility. AB - This study examined the effects of a dopamine D(1) antagonist, SCH23390, infused into the prelimbic-infralimbic areas on the acquisition of a response and visual cue discrimination task, as well as a shift from a response to a visual-cue discrimination and vice versa. Each test was carried out in a cross-maze. The response discrimination required learning to always turn in the same direction (right or left) for a cereal reinforcement. The visual-cue discrimination required learning to always enter the arm with the visual cue. In experiment 1, rats were tested on the response discrimination task, followed by the visual-cue discrimination task. In experiment 2, the testing order was reversed. Bilateral infusions of SCH23390 (0.1 or 1 microg/0.5 microL) into the prelimbic-infralimbic areas did not impair acquisition of the response or visual-cue discrimination tasks. SCH23390 injections at 1 microg, but not 0.1 microg impaired performance when shifting from a response to a visual-cue discrimination, and vice versa. Analysis of the errors revealed that the deficit was due to perseveration of the previously learned strategy. These results suggest that activation of dopamine D(1) receptors in the prelimbic-infralimbic areas may be critical for the suppression of a previously relevant strategy and/or generating new strategies. PMID- 11917004 TI - Interaction between amount and pattern of training in the induction of intermediate- and long-term memory for sensitization in aplysia. AB - In Aplysia, three distinct phases of memory for sensitization can be dissociated based on their temporal and molecular features. A single training trial induces short-term memory (STM, lasting <30 min), whereas five trials delivered at 15-min intervals induces both intermediate-term memory (ITM, lasting >90 min) and long term memory (LTM, lasting >24 h). Here, we explore the interaction of amount and pattern of training in establishing ITM and LTM by examining memory for sensitization after different numbers of trials (each trial = one tail shock) and different patterns of training (massed vs. spaced). Under spaced training patterns, two trials produced STM exclusively, whereas four or five trials each produced both ITM and LTM. Three spaced trials failed to induce LTM but did produce an early decaying form of ITM (E-ITM) that was significantly shorter and weaker in magnitude than the late-decaying ITM (L-ITM) observed after four to five trials. In addition, E-ITM was induced after three trials with both massed and spaced patterns of training. However, L-ITM and LTM after four to five trials require spaced training: Four or five massed trials failed to induce LTM and produced only E-ITM. Collectively, our results indicate that in addition to three identified phases of memory for sensitization--STM, ITM, and LTM--a unique temporal profile of memory, E-ITM, is revealed by varying either the amount or pattern of training. PMID- 11917005 TI - Post-training cyclooxygenase-2 (COX-2) inhibition impairs memory consolidation. AB - Evidence indicates that prostanoids, such as prostaglandins, play a regulatory role in several forms of neural plasticity, including long-term potentiation, a cellular model for certain forms of learning and memory. In these experiments, the significance of the COX isoforms cyclooxygenase-1 (COX-1) and cyclooxygenase 2 (COX-2) in post-training memory processes was assessed. Adult male Long-Evans rats underwent an eight-trial (30-sec intertrial interval) training session on a hippocampus-dependent (hidden platform) or dorsal striatal-dependent (visible platform) tasks in a water maze. After the completion of training, rats received an intraperitoneal injection of the nonselective COX inhibitor indomethacin, the COX-1-specific inhibitor piroxicam, the COX-2-specific inhibitor N-[2 cyclohexyloxy-4-nitrophenyl]-methanesulfonamide (NS-398), vehicle (45% 2 hydroxypropyl-beta-cyclodextrin in distilled water), or saline. On a two-trial retention test session 24 h later, latency to mount the escape platform was used as a measure of memory. In the hidden platform task, the retention test escape latencies of rats administered indomethacin (5 and 10 mg/kg) or NS-398 (2 and 5 mg/kg) were significantly higher than those of vehicle-treated rats, indicating an impairment in retention. Injections of indomethacin or NS-398 that were delayed 2 h post-training had no effect on retention. Post-training indomethacin or NS-398 had no influence on retention of the visible platform version of the water maze at any of the doses administered. Furthermore, selective inhibition of COX-1 via post-training piroxicam administration had no effect on retention of either task. These findings indicate that COX-2 is a required biochemical component mediating the consolidation of hippocampal-dependent memory. PMID- 11917008 TI - A positively charged residue of phi29 DNA polymerase, highly conserved in DNA polymerases from families A and B, is involved in binding the incoming nucleotide. AB - Alignment of the protein sequence of DNA-dependent DNA polymerases has allowed the definition of a new motif, lying adjacent to motif B in the direction of the N-terminus and therefore named pre-motif B. Both motifs are located in the fingers subdomain, shown to rotate towards the active site to form a dNTP-binding pocket in several DNA polymerases in which a closed ternary complex pol:DNA:dNTP has been solved. The functional significance of pre-motif B has been studied by site-directed mutagenesis of phi29 DNA polymerase. The affinity for nucleotides of phi29 DNA polymerase mutant residues Ile364 and Lys371 was strongly affected in DNA- and terminal protein-primed reactions. Additionally, mutations in Ile364 affected the DNA-binding capacity of phi29 DNA polymerase. The results suggest that Lys371 of phi29 DNA polymerase, highly conserved among families A and B, interacts with the phosphate groups of the incoming nucleotide. On the other hand, the role of residue Ile364 seems to be structural, being important for both DNA and dNTP binding. Pre-motif B must therefore play an important role in binding the incoming nucleotide. Interestingly, the roles of Lys371 and Ile364 were also shown to be important in reactions without template, suggesting that phi29 DNA polymerase can achieve the closed conformation in the absence of a DNA template. PMID- 11917009 TI - Physical and functional interactions of the tumor suppressor protein p53 and DNA polymerase alpha-primase. AB - The wild-type form of p53 contains an intrinsic 3'-5'-exonuclease activity. As p53 forms a complex with DNA polymerase alpha-primase (pol-prim) in vivo this finding suggests that p53 might cooperate with pol-prim to stabilize the genetic information of living cells. To test this hypothesis, exonuclease-free DNA pol prim was expressed alone or together with p53 for purification. Pol-prim formed a complex with p53, which was purified by ion exchange and immunoaffinity chromatography from baculovirus-infected insect cells. The p53-containing pol prim fractions removed a 3'-unpaired nucleotide with a 1.5-2-fold higher rate than a paired nucleotide, whereas the four subunit pol-prim did not have any exonuclase activity. Therefore, only p53/pol-prim was able to elongate a primer template that contained a 3'-unpaired primer end in vitro. To achieve this, the 3'-5'-exonuclease activity of p53 excised the unpaired nucleotide at the 3'-end of the primer and created a paired 3'-end, which pol-prim was able to elongate. The exonuclease activity of p53 as well as the elongation of a primer with a mispaired 3'-end was inhibited specifically by the anti-p53 monoclonal antibodies PAb240 and PAb421. PMID- 11917010 TI - NMR structure of a parallel-stranded DNA duplex at atomic resolution. AB - DNA dodecamers have been designed with two cytosines on each end and intervening A and T stretches, such that the oligomers have fully complementary A:T base pairs when aligned in the parallel orientation. Spectroscopic (UV, CD and IR), NMR and molecular dynamics studies have shown that oligomers having the sequences d(CCATAATTTACC) and d(CCTATTAAATCC) form a parallel-stranded duplex when dissolved at 1:1 stoichiometry in aqueous solution. This is due to the C:C+ clamps on either end and extensive mismatches in the antiparallel orientation. The structure is stable at neutral and acidic pH. At higher temperatures, the duplex melts into single strands in a highly cooperative fashion. All adenine, cytosine and thymine nucleotides adopt the anti conformation with respect to the glycosidic bond. The A:T base pairs form reverse Watson-Crick base pairs. The duplex shows base stacking and NOEs between the base protons T(H6)/A(H8) and the sugar protons (H1'/H2'/H2") of the preceding nucleotide, as has been observed in antiparallel duplexes. However, no NOEs are observed between base protons H2/H6/H8 of sequential nucleotides, though such NOEs are observed between T(CH3) and A(H8). A three-dimensional structure of the parallel-stranded duplex at atomic resolution has been obtained using molecular dynamics simulations under NMR constraints. The simulated structures have torsional angles very similar to those found in B-DNA duplexes, but the base stacking and helicoid parameters are significantly different. PMID- 11917011 TI - Receptor-mediated endocytosis of phosphodiester oligonucleotides in the HepG2 cell line: evidence for non-conventional intracellular trafficking. AB - Having identified an oligonucleotide (ON) receptor in the HepG2 cell line, we have re-examined here the kinetics of ON uptake, subcellular distribution and intracellular localisation in these cells, at concentrations relevant for the study of a receptor-dependent process. Kinetic parameters of ON endocytosis were comparable with those of the receptor-mediated endocytosis tracer, transferrin (uptake equilibrium, saturation with concentration, specific competition and rapid efflux) and were clearly distinct from those of fluid-phase endocytosis. By analytical subcellular fractionation, particulate ON showed a bimodal distribution after 2 h of uptake, with a low-density peak superimposed on the distribution of endosomes, and a high-density peak overlapping lysosomes. After an overnight chase, only the high-density peak remained, but it could be dissociated from lysosomes, based on its refractoriness to displacement upon chloroquine-induced swelling. After 2 h of uptake at 300 nM ON-Alexa, a punctate pattern was resolved, by confocal microscopy, from those of transferrin, of a fluid-phase tracer, and of vital staining of lysosomes by LysoTracker. At 3 microM ON-Alexa, its pattern largely overlapped with the fluid-phase tracer and LysoTracker. Taken together, these data suggest that ON may be internalised at low concentrations by receptor-mediated endocytosis into unique endosomes, then to dense structures that are distinct from lysosomes. The nature of these two compartments and their significance for ON effect deserve further investigation. PMID- 11917006 TI - Comparative genomics and evolution of proteins involved in RNA metabolism. AB - RNA metabolism, broadly defined as the compendium of all processes that involve RNA, including transcription, processing and modification of transcripts, translation, RNA degradation and its regulation, is the central and most evolutionarily conserved part of cell physiology. A comprehensive, genome-wide census of all enzymatic and non-enzymatic protein domains involved in RNA metabolism was conducted by using sequence profile analysis and structural comparisons. Proteins related to RNA metabolism comprise from 3 to 11% of the complete protein repertoire in bacteria, archaea and eukaryotes, with the greatest fraction seen in parasitic bacteria with small genomes. Approximately one-half of protein domains involved in RNA metabolism are present in most, if not all, species from all three primary kingdoms and are traceable to the last universal common ancestor (LUCA). The principal features of LUCA's RNA metabolism system were reconstructed by parsimony-based evolutionary analysis of all relevant groups of orthologous proteins. This reconstruction shows that LUCA possessed not only the basal translation system, but also the principal forms of RNA modification, such as methylation, pseudouridylation and thiouridylation, as well as simple mechanisms for polyadenylation and RNA degradation. Some of these ancient domains form paralogous groups whose evolution can be traced back in time beyond LUCA, towards low-specificity proteins, which probably functioned as cofactors for ribozymes within the RNA world framework. The main lineage-specific innovations of RNA metabolism systems were identified. The most notable phase of innovation in RNA metabolism coincides with the advent of eukaryotes and was brought about by the merge of the archaeal and bacterial systems via mitochondrial endosymbiosis, but also involved emergence of several new, eukaryote-specific RNA-binding domains. Subsequent, vast expansions of these domains mark the origin of alternative splicing in animals and probably in plants. In addition to the reconstruction of the evolutionary history of RNA metabolism, this analysis produced numerous functional predictions, e.g. of previously undetected enzymes of RNA modification. PMID- 11917007 TI - Transcriptional silencing in Saccharomyces cerevisiae and Schizosaccharomyces pombe. AB - Transcriptional silencing is a heritable form of gene inactivation that involves the assembly of large regions of DNA into a specialized chromatin structure that inhibits transcription. This phenomenon is responsible for inhibiting transcription at silent mating-type loci, telomeres and rDNA repeats in both budding yeast Saccharomyces cerevisiae and fission yeast Schizosaccharomyces pombe, as well as at centromeres in fission yeast. Although transcriptional silencing in both S.cerevisiae and S.pombe involves modification of chromatin, no apparent amino acid sequence similarities have been reported between the proteins involved in establishment and maintenance of silent chromatin in these two distantly related yeasts. Silencing in S.cerevisiae is mediated by Sir2p containing complexes, whereas silencing in S.pombe is mediated primarily by Swi6 containing complexes. The Swi6 complexes of S.pombe contain proteins closely related to their counterparts in higher eukaryotes, but have no apparent orthologs in S.cerevisiae. Silencing proteins from both yeasts are also actively involved in other chromosome-related nuclear functions, including DNA repair and the regulation of chromatin structure. PMID- 11917012 TI - Requirements for utilization of CREB binding protein by hypersensitive site two of the beta-globin locus control region. AB - Strong transactivation of the beta-globin genes is conferred by the beta-globin locus control region (LCR), which consists of four erythroid-specific DNase I hypersensitive sites (HS1-HS4). HS2 has a powerful enhancer activity dependent upon tandem binding sites for the erythroid cell- and megakaryocyte-specific transcription factor NF-E2. An important co-activator-mediating transactivation by HS2 is the histone acetyltransferase (HAT) CREB binding protein (CBP). We showed previously that recruitment of a GAL4-CBP fusion protein to HS2 largely bypassed the requirement of the NF-E2 sites for transactivation. To determine whether GAL4-CBP recruitment is sufficient for transactivation, we assessed the importance of cis-elements within HS2. Docking of GAL4-CBP upstream of an Agamma globin promoter lacking HS2 only weakly activated the promoter, indicating that HS2 components are required for GAL4-CBP-mediated transactivation. Sequences upstream and downstream of the NF-E2 sites were required for maximal GAL4-CBP mediated transactivation, and HAT catalytic activity of GAL4-CBP was critical. No single factor-binding site was required for GAL4-CBP-mediated transactivation. However, deletion of two sites, a CACC site and an E-box, abolished transactivation in transient and stable transfection assays. These results suggest that NF-E2 recruits CBP as a critical step in transactivation, but additional components of HS2 are required to achieve maximal enhancer activity. PMID- 11917013 TI - Correlated alternative side chain conformations in the RNA-recognition motif of heterogeneous nuclear ribonucleoprotein A1. AB - The RNA-recognition motif (RRM) is a common and evolutionarily conserved RNA binding module. Crystallographic and solution structural studies have shown that RRMs adopt a compact alpha/beta structure, in which four antiparallel beta strands form the major RNA-binding surface. Conserved aromatic residues in the RRM are located on the surface of the beta-sheet and are important for RNA binding. To further our understanding of the structural basis of RRM-nucleic acid interaction, we carried out a high resolution analysis of UP1, the N-terminal, two-RRM domain of heterogeneous nuclear ribonucleoprotein A1 (hnRNP A1), whose structure was previously solved at 1.75-1.9 A resolution. The two RRMs of hnRNP A1 are closely related but have distinct functions in regulating alternative pre mRNA splice site selection. Our present 1.1 A resolution crystal structure reveals that two conserved solvent-exposed phenylalanines in the first RRM have alternative side chain conformations. These conformations are spatially correlated, as the individual amino acids cannot adopt each of the observed conformations independently. These phenylalanines are critical for nucleic acid binding and the observed alternative side chain conformations may serve as a mechanism for regulating nucleic acid binding by RRM-containing proteins. PMID- 11917014 TI - Rhodobacter sphaeroides LexA has dual activity: optimising and repressing recA gene transcription. AB - Transcription of the Rhodobacter sphaeroides recA promoter (P(recA)) is induced upon DNA damage in a lexA-dependent manner. In vivo experiments demonstrate that LexA protein represses and might also activate transcription of P(recA). Purified R.sphaeroides LexA protein specifically binds the SOS boxes located within the P(recA) region. In vitro transcription analysis, using Escherichia coli RNA polymerase (RNAP), indicated that the presence of LexA may stimulate and repress transcription of P(recA). EMSA and DNase I footprinting experiments show that LexA and RNAP can bind simultaneously to P(recA). At low LexA concentrations it enhances RNAP binding to P(recA), stimulates open complex formation and strand separation beyond the transcription start site. At high LexA concentrations, however, RNAP-promoted strand separation is not observed beyond the +5 region. LexA might repress transcription by interfering with the clearance process instead of blocking the access of RNAP to the promoter region. Based on these findings we propose that the R.sphaeroides LexA protein performs fine tuning of the SOS response, which might provide a physiological advantage by enhancing transcription of SOS genes and delaying full activation of the response. PMID- 11917015 TI - Circular permutation of DNA cytosine-N4 methyltransferases: in vivo coexistence in the BcnI system and in vitro probing by hybrid formation. AB - Sequence analysis of the BcnI restriction-modification system from Bacillus centrosporus revealed four open reading frames (bcnIC, bcnIR, bcnIB and bcnIA) that are arranged as two converging collinear pairs. One pair encodes a putative small regulatory protein, C.BcnI, and the restriction endonuclease R.BcnI. The other two gene products are the DNA cytosine-N4 methyltransferases M.BcnIA and M.BcnIB, which differ by circular permutation of conserved sequence motifs. The BcnI methyltransferases are isospecific on double-stranded DNA [methylation specificity CC(C/G)GG], but M.BcnIA can also methylate the target sites in single stranded DNA. Functional analysis shows that bcnIA is dispensable (bcnIB is capable of protecting the DNA against the in vivo activity of bcnIR); in contrast, no stable clones were obtained if bcnIB alone was deleted from the system. By analogy with the DpnII system, the second methylase M.BcnIA may play a role in the transformation proficiency of its gram-positive host. The interchangeability of homologous elements in the beta class of cytosine-N4 methylases was probed by hybrid formation between M.BcnIB and its closest homolog M.Cfr9I (CCCGGG) employing a novel semi-random strategy combined with selection for catalytic activity. The fusion points in the active hybrids mapped in a narrow region located between sequence motifs X and I. Our data illustrate that recombination of two related sequences by circular permutation may serve as an evolutionary mechanism for creating new specificities of amino MTases. PMID- 11917016 TI - Multiple thiol-anchor capped DNA-gold nanoparticle conjugates. AB - We report the synthesis of a novel trithiol-capped oligodeoxyribonucleotide and gold nanoparticle conjugates prepared from it. These DNA-gold nanoparticle conjugates exhibit substantially higher stability than analogs prepared from monothiol and cyclic disulfide-capped oligodeoxyribonucleotides, but comparable hybridization properties. A quantitative analysis of their stability under a range of conditions is provided. Significantly, this novel trithiol oligodeoxyribonucleotide can be used to stabilize particles >30 nm in diameter, which are essential for many diagnostic applications. PMID- 11917018 TI - An efficient algorithm for large-scale detection of protein families. AB - Detection of protein families in large databases is one of the principal research objectives in structural and functional genomics. Protein family classification can significantly contribute to the delineation of functional diversity of homologous proteins, the prediction of function based on domain architecture or the presence of sequence motifs as well as comparative genomics, providing valuable evolutionary insights. We present a novel approach called TRIBE-MCL for rapid and accurate clustering of protein sequences into families. The method relies on the Markov cluster (MCL) algorithm for the assignment of proteins into families based on precomputed sequence similarity information. This novel approach does not suffer from the problems that normally hinder other protein sequence clustering algorithms, such as the presence of multi-domain proteins, promiscuous domains and fragmented proteins. The method has been rigorously tested and validated on a number of very large databases, including SwissProt, InterPro, SCOP and the draft human genome. Our results indicate that the method is ideally suited to the rapid and accurate detection of protein families on a large scale. The method has been used to detect and categorise protein families within the draft human genome and the resulting families have been used to annotate a large proportion of human proteins. PMID- 11917017 TI - Factors governing loss and rescue of DNA binding upon single and double mutations in the p53 core domain. AB - The mutation of R273-->H in the p53 core domain (p53-CD) is one of the most common mutations found in human cancers. Although the 273H p53-CD retains the wild-type conformation and stability, it lacks sequence-specific DNA binding, a transactivation function and growth suppression. However, mutating T284-->R in the 273H p53-CD restores the DNA binding affinity, and transactivation and tumour suppressor functions. Since X-ray/NMR structures of DNA-free or DNA-bound mutant p53-CD molecules are unavailable, the factors governing the loss and rescue of sequence-specific DNA binding in the 273H and 273H+284R p53-CD, respectively, are unclear. Hence, we have carried out molecular dynamics (MD) simulations of the wild-type, single mutant and double mutant p53-CD, free and DNA bound, in the presence of explicit water molecules. Based on the MD structures, the DNA-binding free energy of each p53 molecule has been computed and decomposed into component energies and contributions from the interface residues. The wild-type and mutant p53-CD MD structures were found to be consistent with the antibody-binding, X-ray and NMR data. The predicted DNA binding affinity and specificity of both mutant p53-CDs were also in accord with experimental data. The non-detectable DNA binding of the 273H p53-CD is due mainly to the disruption of a hydrogen-bonding network involving R273, D281 and R280, leading to a loss of major groove binding by R280 and K120. The restoration of DNA binding affinity and specificity of the 273H+284R p53-CD is due mainly to the introduction of another DNA-binding site at position 284, leading to a recovery of major groove binding by R280 and K120. The important role of water molecules and the DNA major groove conformation as well as implications for structure-based linker rescue of the 273H p53-CD DNA-binding affinity are discussed. PMID- 11917019 TI - A novel DNA polymerase homologous to Escherichia coli DNA polymerase I from a higher plant, rice (Oryza sativa L.). AB - A novel DNA polymerase, designated as OsPolI-like, has been identified from the higher plant, rice (Oryza sativa L. cv. Nipponbare). The OsPolI-like cDNA was 3765 bp in length, and the open reading frame encoded a predicted product of 977 amino acid residues with a molecular weight of 100 kDa. The OsPolI-like gene has been mapped to chromosome 8 and contains 12 exons and 11 introns. The encoded protein showed a high degree of sequence and structural homology to Escherichia coli pol I protein, but differed from DNA polymerase gamma and theta. The DNA polymerase domain of OsPolI-like showed DNA polymerase activity. Subcellular fractionation analysis suggested that the protein is localized in the plastid. Northern and western blotting, and in situ hybridization analyses demonstrated preferential expression of OsPolI-like in meristematic tissues such as shoot apical meristem, root apical meristem, leaf primordia and the marginal meristem. Interestingly, no expression was detected in mature leaves, although they have a high chloroplast content. These properties indicated that OsPolI-like is a novel plant DNA polymerase. The function of OsPolI-like is discussed in relation to plastid maturation. PMID- 11917020 TI - Restart of DNA replication in Gram-positive bacteria: functional characterisation of the Bacillus subtilis PriA initiator. AB - The PriA protein was identified in Escherichia coli as a factor involved in the replication of extrachromosomal elements such as bacteriophage phiX174 and plasmid pBR322. Recent data show that PriA plays an important role in chromosomal replication, by promoting reassembly of the replication machinery during reinitiation of inactivated forks. A gene encoding a product 32% identical to the E.coli PriA protein has been identified in Bacillus subtilis. To characterise this protein, designated PriA(Bs), we constructed priA(Bs) mutants. These mutants are poorly viable, filamentous and sensitive to rich medium and UV irradiation. Replication of pAMbeta1-type plasmids, which is initiated through the formation of a D-loop structure, and the activity of the primosome assembly site ssiA of plasmid pAMbeta1 are strongly affected in the mutants. The purified PriA(Bs) protein binds preferentially to the active strand of ssiA, even in the presence of B.subtilis SSB protein (SSB(Bs)). PriA(Bs) also binds stably and specifically to an artificial D-loop structure in vitro. These data show that PriA(Bs) recognises two specific substrates, ssiA and D-loops, and suggest that it triggers primosome assembly on them. PriA(Bs) also displays a single-stranded DNA dependent ATPase activity, which is reduced in the presence of SSB(Bs), unless the ssiA sequence is present on the ssDNA substrate. Finally, PriA(Bs) is shown to be an active helicase. Altogether, these results demonstrate a clear functional identity between PriA(Ec) and PriA(Bs). However, PriA(Bs) does not complement an E.coli priA null mutant strain. This host specificity may be due to the divergence between the proteins composing the E.coli and B.subtilis PriA dependent primosomes. PMID- 11917021 TI - Efficient trans-cleavage by the Schistosoma mansoni SMalpha1 hammerhead ribozyme in the extreme thermophile Thermus thermophilus. AB - The catalytic hammerhead structure has been found in association with repetitive DNA from several animals, including salamanders, crickets and schistosomes, and functions to process in cis the long multimer transcripts into monomer RNA in vivo. The cellular role of these repetitive elements and their transcripts is unknown. Moreover, none of these natural hammerheads have been shown to trans cleave a host mRNA in vivo. We analyzed the cis- and trans-cleavage properties of the hammerhead ribozyme associated with the SMalpha DNA family from the human parasite Schistosoma mansoni. The efficiency of trans-cleavage of a target RNA in vitro was affected mainly by both the temperature-dependent chemical step and the ribozyme-product dissociation step. The optimal temperature for trans-cleavage was 70 degrees C. This result was confirmed when both the SMalpha1 ribozyme and the target RNA were expressed in the extreme thermophile Thermus thermophilus. Moreover, SMalpha1 RNA showed a remarkable thermostability, equal or superior to that of the most stable RNAs in this species, suggesting that SMalpha1 RNA has been selected for stability. Computer analysis predicts that the monomer and multimer transcripts fold into highly compact secondary structures, which may explain their exceptional stability in vivo. PMID- 11917022 TI - Immunostimulatory properties of phosphorothioate CpG DNA containing both 3'-5'- and 2'-5'-internucleotide linkages. AB - Synthetic oligodeoxyribonucleotides containing CpG-dinucleotides (CpG DNA) in specific sequence contexts activate the vertebrate immune system. We have examined the effect of 3'-deoxy-2'-5'-ribonucleoside (3'-deoxynucleoside) incorporation into CpG DNA on the immunostimulatory activity. Incorporation of 3' deoxynucleosides results in the formation of 2'-5'-internucleotide linkages in an otherwise 3'-5'-linked CpG DNA. In studies, both in vitro and in vivo, CpG DNA containing unnatural 3'-deoxynucleoside either within the CpG-dinucleotide or adjacent to the CpG-dinucleotide failed to induce immunostimulatory activity, suggesting that the modification was not recognized by the receptors. Incorporation of the same modification distal to the CpG-dinucleotide in the 5' flanking sequence potentiated the immunostimulatory activity of the CpG DNA. The same modification when incorporated in the 3'-flanking sequence had an insignificant effect on immunostimulatory activity of CpG DNA. Interestingly, substitution of a 3'-deoxynucleoside in the 5'-flanking sequence distal to the CpG-dinucleotide resulted in increased IL-6 and IL-10 secretion with similar levels of IL-12 compared with parent CpG DNA. The incorporation of the same modification in the 3'-flanking sequence resulted in lower IL-6 and IL-10 secretion with similar levels of IL-12 compared with parent CpG DNA. These results suggest that site-specific incorporation of 3'-deoxynucleotides in CpG DNA modulates immunostimulatory properties. PMID- 11917023 TI - SmpB functions in various steps of trans-translation. AB - tmRNA has a dual function as a tRNA and an mRNA to facilitate trans-translation, in which a ribosome can switch between translation of a truncated mRNA and the tmRNA's tag sequence. SmpB is a tmRNA binding protein that has been identified to be essential for trans-translation in vivo. To further study the function of SmpB, an S30 fraction from an Escherichia coli strain, in which the set of genes for SmpB and tmRNA has been deleted from the genome, and His-tagged SmpB active in trans-translation were prepared. The SmpB-depleted S30 fraction had an ability to facilitate poly(U)-dependent tag-peptide synthesis in vitro when purified His tagged SmpB was exogenously added together with tmRNA, although SmpB was not required for in vitro poly(U)-dependent poly(Phe) synthesis. It was also found that depletion of SmpB leads to a decrease in the level of tmRNA in the cell. In addition, SmpB considerably enhanced the aminoacylation of tmRNA by alanyl-tRNA synthetase in vitro. The aminoacylation enhancement by SmpB, the binding of SmpB to tmRNA and the effect of depletion of SmpB on the expression level of tmRNA in the cell were all affected by some mutations in the tRNA-like domain which cause a defect in ribosome binding leading to a trans-translation deficiency. These results demonstrate that, via binding to the tRNA-like domain of tmRNA, SmpB plays various roles: rescuing the tmRNA molecule from degradation in the cell, enhancing the aminoacylation of tmRNA and mediating the binding of tmRNA to ribosome. PMID- 11917024 TI - Response of human REV1 to different DNA damage: preferential dCMP insertion opposite the lesion. AB - REV1 functions in the DNA polymerase zeta mutagenesis pathway. To help understand the role of REV1 in lesion bypass, we have examined activities of purified human REV1 opposite various template bases and several different DNA lesions. Lacking a 3'-->5' proofreading exonuclease activity, purified human REV1 exhibited a DNA polymerase activity on a repeating template G sequence, but catalyzed nucleotide insertion with 6-fold lower efficiency opposite a template A and 19-27-fold lower efficiency opposite a template T or C. Furthermore, dCMP insertion was greatly preferred regardless of the specific template base. Human REV1 inserted a dCMP efficiently opposite a template 8-oxoguanine, (+)-trans-anti-benzo[a]pyrene-N2 dG, (-)-trans-anti-benzo[a]pyrene-N2-dG and 1,N6-ethenoadenine adducts, very inefficiently opposite an acetylaminofluorene-adducted guanine, but was unresponsive to a template TT dimer or TT (6-4) photoproduct. Surprisingly, the REV1 specificity of nucleotide insertion was very similar in response to different DNA lesions with greatly preferred C insertion and least frequent A insertion. By combining the dCMP insertion activity of human REV1 with the extension synthesis activity of human polymerase kappa, bypass of the trans-anti benzo[a]pyrene-N2-dG adducts and the 1,N6-ethenoadenine lesion was achieved by the two-polymerase two-step mechanism. These results suggest that human REV1 is a specialized DNA polymerase that may contribute to dCMP insertion opposite many types of DNA damage during lesion bypass. PMID- 11917025 TI - Preparation of partially 2H/13C-labelled RNA for NMR studies. Stereo-specific deuteration of the H5" in nucleotides. AB - An effective in vitro enzymatic synthesis is described for the production of nucleoside triphosphates (NTPs) which are stereo-specifically deuterated on the H5" position with high selectivity (>98%), and which can have a variety of different labels (13C, 15N, 2H) in other positions. The NTPs can subsequently be employed in the enzymatic synthesis of RNAs using T7 polymerase from a DNA template. The stereo-specific deuteration of the H5" immediately provides the stereo-specific assignment of H5' resonances in NMR spectra, giving access to important structural parameters. Stereo-chemical H-exchange was used to convert commercially available 1,2,3,4,5,6,6-2H-1,2,3,4,5,6-13C-D-glucose (d7-13C6-D glucose) into [1,2,3,4,5,6(R)-2H-1,2,3,4,5,6-13C]-D-glucose (d6-13C6-D-glucose). [1',3',4',5"-2H-1',2',3',4',5'-13C]GTP (d4-13C5-GTP) was then produced from d6 13C6-D-glucose and guanine base via in vitro enzymatic synthesis employing enzymes from the pentose-phosphate, nucleotide biosynthesis and salvage pathways. The overall yield was approximately 60 mg NTP per 1 g glucose, comparable with the yield of NTPs isolated from Escherichia coli grown on enriched media. The d4 13C5-GTP, together with in vitro synthesised d5-UTP, d5-CTP and non-labelled ATP, were used in the synthesis of a 31 nt RNA derived from the primer binding site of hepatitis B virus genomic RNA. (13C,1H) hetero-nuclear multiple-quantum spectra of the specifically deuterated sample and of a non-deuterated uniformly 13C/15N labelled sample demonstrates the reduced spectral crowding and line width narrowing compared with 13C-labelled non-deuterated RNA. PMID- 11917026 TI - Sequence requirements for terminators and antiterminators in the T box transcription antitermination system: disparity between conservation and functional requirements. AB - The T box transcription termination control system is used in Gram-positive bacteria to regulate expression of aminoacyl-tRNA synthetase and other amino acid related genes. Readthrough of a transcriptional terminator located in the leader region of the target gene is dependent on a specific interaction between the nascent leader transcript and the cognate uncharged tRNA. This interaction is required for formation of an antiterminator structure in the leader, which prevents formation of a competing transcriptional terminator stem-loop. The antiterminators and terminators of genes in this family are highly conserved in both secondary structure and primary sequence; the antiterminator contains the T box sequence, which is the most highly conserved leader element. These conserved features were investigated by phylogenetic and mutational analysis. Changes at highly conserved positions in the bulge and in the helix above the bulge reduced function, while alteration of other positions that were as much as 96% conserved did not have a major effect. The disparity between sequence conservation and function may be due to the requirement for maintaining base pairing in both the antiterminator and terminator structures. PMID- 11917027 TI - A dual reporter screening system identifies the amino acid at position 82 in Flp site-specific recombinase as a determinant for target specificity. AB - We have developed a dual reporter screen in Escherichia coli for identifying variants of the Flp site-specific recombinase that have acquired reactivity at an altered target site (mFRT). In one reporter, the lacZalpha gene segment is flanked by mFRTs in direct orientation. In the other, the red fluorescence protein (RFP) gene is flanked by the native FRTs. Hence, the color of a colony on an X-gal indicator plate indicates the recombination potential of the variant Flp protein expressed in it: blue if no recombination or only FRT recombination occurs, red if only mFRT recombination occurs and white if both FRT and mFRT recombinations occur. The scheme was validated by identification and in vivo characterization of Flp variants that show either relaxed specificity (active on FRT and mFRT) or moderately shifted specificity toward mFRT. We find that alteration of Lys-82 to Met, Thr, Arg or His enables the corresponding Flp variants to recombine FRT sites as well as altered FRT sites containing a substitution of G-C by C-G at position 1 of the Flp binding element (mFRT11). In contrast, wild-type Flp has no detectable activity on mFRT11. When Lys-82 is replaced by Tyr, the resulting Flp variant shows a small but reproducible preference for mFRT11 over FRT. However, this preference for mFRT11 is nearly lost when Tyr-82 is substituted by Phe. PMID- 11917028 TI - Post-transcriptional suppression of gene expression in Xenopus embryos by small interfering RNA. AB - Double-stranded RNA (dsRNA) induces gene-specific silencing in organisms from fungi to animals, a phenomenon known as RNA interference (RNAi). RNAi represents an evolutionarily conserved system to protect against aberrant expression of genes and a powerful tool for gene manipulation. Despite reports that RNAi can be induced in vertebrates, severe sequence-non-specific effects of long dsRNA have been documented in various systems. It has recently been shown in cultured mammalian cells that small interfering RNAs (siRNAs) of 21-23 nt can mediate RNAi but bypass the non-specific response induced by longer dsRNAs. However, the effectiveness of siRNAs has not been demonstrated in living vertebrates. In addition, the mechanism of siRNA suppression of gene expression in vertebrate cells remains to be elucidated. Here we show that synthetic 21 nt siRNAs can specifically inhibit the expression of exogenously introduced as well as endogenous genes in the embryos of Xenopus laevis. siRNAs significantly reduced the steady-state amount of both the mRNA and protein of the cognate gene target. Moreover, co-injection of siRNA with the target RNA transcript specifically suppressed the activity of the latter. Taken together, our findings establish siRNA-mediated post-transcriptional suppression of gene expression in Xenopus embryos. PMID- 11917029 TI - The zinc ion in the HNH motif of the endonuclease domain of colicin E7 is not required for DNA binding but is essential for DNA hydrolysis. AB - The HNH motif was originally identified in the subfamily of HNH homing endonucleases, which initiate the process of the insertion of mobile genetic elements into specific sites. Several bacteria toxins, including colicin E7 (ColE7), also contain the 30 amino acid HNH motif in their nuclease domains. In this work, we found that the nuclease domain of ColE7 (nuclease-ColE7) purified from Escherichia coli contains a one-to-one stoichiometry of zinc ion and that this zinc-containing enzyme hydrolyzes DNA without externally added divalent metal ions. The apo-enzyme, in which the indigenous zinc ion was removed from nuclease-ColE7, had no DNase activity. Several divalent metal ions, including Ni2+, Mg2+, Co2+, Mn2+, Ca2+, Sr2+, Cu2+ and Zn2+, re-activated the DNase activity of the apo-enzyme to various degrees, however higher concentrations of zinc ion inhibited this DNase activity. Two charged residues located at positions close to the zinc-binding site were mutated to alanine. The single-site mutants, R538A and E542A, showed reduced DNase activity, whereas the double-point mutant, R538A + E542A, had no observable DNase activity. A gel retardation assay further demonstrated that the nuclease-ColE7 hydrolyzed DNA in the presence of zinc ions, but only bound to DNA in the absence of zinc ions. These results demonstrate that the zinc ion in the HNH motif of nuclease-ColE7 is not required for DNA binding, but is essential for DNA hydrolysis, suggesting that the zinc ion not only stabilizes the folding of the enzyme, but is also likely to be involved in DNA hydrolysis. PMID- 11917030 TI - dNTP versus NTP discrimination by phenylalanine 451 in duck hepatitis B virus P protein indicates a common structure of the dNTP-binding pocket with other reverse transcriptases. AB - Hepatitis B viruses, or hepadnaviruses, are small DNA-containing viruses that replicate through reverse transcription. Their prototype, HBV, causes severe liver disease in humans. The hepadnaviral P protein is an unusual reverse transcriptase (RT) that initiates DNA synthesis by host-factor-dependent protein priming on a specific RNA stem-loop template, epsilon, yielding a short DNA oligonucleotide covalently attached to the RT. This priming reaction can be reconstituted with in vitro-translated duck hepatitis B virus (DHBV) P protein. No direct structural data are available for any P protein. However, P proteins share a number of conserved motifs with other polymerases. Box A contains an invariant bulky residue recently shown to be crucial for dNTP versus NTP discrimination in RTs and some DNA polymerases; its equivalent in DHBV P protein would be phenylalanine 451 (F451). Four mutants, containing glycine (F451G), alanine (F451A), valine (F451V) and aspartate (F451D), were therefore analyzed for their ability to utilize dNTPs and NTPs in in vitro priming. Priming efficiencies with dNTPs decreased with decreasing side chain size but GTP utilization increased; the wild-type enzyme was inactive with GTP. In the context of complete DHBV genomes, all mutant proteins were competent for RNA encapsidation, indicating the absence of global structural alterations. Because the function of the discriminatory residue depends on its specific spatial disposition this strongly suggests a similar architecture for the P protein dNTP binding pocket as in other RTs. PMID- 11917031 TI - Induction of RNA interference in Caenorhabditis elegans by RNAs derived from plants exhibiting post-transcriptional gene silencing. AB - The term 'gene silencing' refers to transcriptional and post-transcriptional control of gene expression. Related processes are found across kingdoms in plants and animals. We intended to test whether particular RNA constituents of a silenced plant can induce silencing in an animal. We generated Nicotiana benthamiana lines that expressed green fluorescent protein (GFP) from a transgene. Plants in which GFP expression was spontaneously silenced showed siRNAs characteristic of post-transcriptional gene silencing (PTGS). RNA extracts prepared from silenced plants were injected into a GFP-expressing strain of Caenorhabditis elegans, where they induced RNA interference (RNAi). Extracts from non-silenced plants were inactive. This directly demonstrates a relationship and a mechanistic link between PTGS and RNAi. Controls confirmed that the silencing agent was an RNA. Size fractionation on denaturing gels revealed that an RNA of approximately 85 nt was most active in inducing silencing in the worm. Northern blot analysis of the region in question did not detect a prominent GFP-specific RNA of sense or antisense polarity, indicating that the RNA species which induced silencing was present only in low concentration or did not hybridize due to formation of an intramolecular double strand. In view of its high activity, it is possible that this agent is responsible for the systemic spread of silencing in plants and it might represent the aberrant RNA, a previously postulated inducer of silencing. PMID- 11917032 TI - The terminase subunits pUL56 and pUL89 of human cytomegalovirus are DNA metabolizing proteins with toroidal structure. AB - Herpesvirus DNA packaging involves binding and cleavage of DNA containing the specific DNA-packaging motifs. Here we report a first characterization of the terminase subunits pUL56 and pUL89 of human cytomegalovirus (HCMV). Both gene products were shown to have comparable nuclease activities in vitro. Under limiting protein concentrations the nuclease activity is enhanced by interaction of pUL56 and pUL89. High amounts of 2-bromo-5,6-dichloro-1-beta-D-ribofuranosyl benzimidazole partially inhibited the pUL89-associated nuclease activity. It was demonstrated that pUL56 is able to bind to nucleocapsids in vivo. Electron microscopy (EM) and image analysis of purified pUL56 revealed that the molecules occurred as a distinct ring-shaped structure with a pronounced cleft. EM analysis of purified pUL89 demonstrated that this protein is also a toroidal DNA metabolizing protein. Upon interaction of pUL56 with linearized DNA, the DNA remains uncut while the cutting event itself is mediated by pUL89. Using biochemical assays in conjunction with EM pUL56 was shown to (i) bind to DNA and (ii) associate with the capsid. In contrast to this, EM analysis implied that pUL89 is required to effect DNA cleavage. The data provide the first insights into the terminase-dependent viral DNA-packaging mechanism of HCMV. PMID- 11917034 TI - Methylome profiling of cancer cells by amplification of inter-methylated sites (AIMS). AB - Alterations of the DNA methylation pattern have been related to generalized chromosomal disruption and inactivation of multiple tumor suppressor genes in neoplasia. To screen for tumor-specific alterations and to make a global assessment of methylation status in cancer cells, we have modified the methylated CpG island amplification method to generate easily readable fingerprints representing the cell's DNA methylation profile. The method is based on the differential cleavage of isoschizomers with distinct methylation sensitivity. Specific adaptors are ligated to the methylated ends of the digested genomic DNA. The ligated sequences are amplified by PCR using adaptor- specific primers extended at the 3' end with two to four arbitrarily chosen nucleotidic residues to reduce the complexity of the product. Fingerprints consist of multiple anonymous bands, representing DNA sequences flanked by two methylated sites, which can be isolated and individually characterized. Hybridization of the whole product to metaphase chromosomes revealed that most bands originate from the isochore H3, which identifies the regions of the genome with the highest content of CpG islands and genes. Comparison of the fingerprints obtained from normal colon mucosa, colorectal carcinomas and cell lines revealed tumor-specific alterations that are putative recurrent markers of the disease and include tumor specific hypo- and hypermethylations. PMID- 11917033 TI - Structural analysis of conserved base pairs in protein-DNA complexes. AB - Understanding of protein-DNA interactions is crucial for prediction of DNA binding specificity of transcription factors and design of novel DNA-binding proteins. In this paper we develop a novel approach to analysis of protein-DNA interactions. We bring together two sources of information: (i) structures of protein-DNA complexes (PDB/NDB database) and (ii) experimentally obtained sites recognized by DNA-binding proteins. Sites are used to compute conservation (information content) of each base pair, which indicates relative importance of the base pair in specific recognition. The main result of this study is that conservation of base pairs in a site exhibits significant correlation with the number of contacts the base pairs have with the protein. In particular, base pairs that have more contacts with the protein are more conserved in evolution. As natural as it is, this result has never been reported before. We also observe that for most of the studied proteins, hydrogen bonds and hydrophobic interactions alone cannot explain the pattern of evolutionary conservation in the binding site suggesting cumulative contribution of different types of interactions to specific recognition. Implications for prediction of the DNA binding specificity are discussed. PMID- 11917035 TI - An in vivo selection system for homing endonuclease activity. AB - Homing endonucleases are enzymes that catalyze the highly sequence-specific cleavage of DNA. We have developed an in vivo selection in Escherichia coli that links cell survival with homing endonuclease-mediated DNA cleavage activity and sequence specificity. Using this selection, wild-type and mutant variants of three homing endonucleases were characterized without requiring protein purification and in vitro analysis. This selection system may facilitate the study of sequence-specific DNA cleaving enzymes, and selections based on this work may enable the evolution of homing endonucleases with novel activities or specificities. PMID- 11917036 TI - An assessment of Motorola CodeLink microarray performance for gene expression profiling applications. AB - DNA microarrays enable users to obtain information on differences in transcript abundance on a massively parallel scale. Recently, however, data analyses have revealed potential pitfalls related to image acquisition, variability and misclassifications in replicate measurements, cross-hybridization and sensitivity limitations. We have generated a series of analytical tools to address the manufacturing, detection and data analysis components of a microarray experiment. Together, we have used these tools to optimize performance in an expression profiling study. We demonstrate three significant advantages of the Motorola CodeLink platform: sensitivity of one copy per cell, coefficients of variation of 10% in the hybridization signals across slides and across target preparations, and specificity in distinguishing highly homologous sequences. Slides where oligonucleotide probes are spotted in 6-fold redundancy were used to demonstrate the effect of replication on data quality. Lastly, the differential expression ratios obtained with the CodeLink expression platform were validated against those obtained with quantitative reverse transcription-PCR assays for 54 genes. PMID- 11917037 TI - Multiplex Pyrosequencing. AB - We describe here the development of a new and simple single-tube multiplex Pyrosequencing assay. Genomic DNA or cDNA was employed to PCR amplify region(s) using biotinylated and normal primer(s). Subsequent to capture of PCR products on streptavidin-coated beads, single-stranded DNA separation and hybridization of multiple sequencing primers, Pyrosequencing was performed. The obtained pyrogram resulted in a unique pattern in which the intensity of the signal determined the number of incorporated nucleotide(s). Here, we demonstrate the use of this multiplex Pyrosequencing for single nucleotide polymorphisms genotyping and microbial typing. PMID- 11917039 TI - TspGWI, a thermophilic class-IIS restriction endonuclease from Thermus sp., recognizes novel asymmetric sequence 5'-ACGGA(N11/9)-3'. AB - A novel prototype class-IIS restriction endonuclease, TspGWI, was isolated from the thermophilic bacterium Thermus sp. GW. The recognition sequence and cleavage positions have been established: TspGWI recognizes the non-palindromic 5-bp sequence 5'-ACGGA-3' and cleaves the DNA 11 and 9 nt downstream in the top and bottom strand, respectively. In addition, an accompanying endonuclease, TspGWII, an isoschizomer of Pst I, was found in Thermus sp. GW cells. PMID- 11917038 TI - Glass bead purification of plasmid template DNA for high throughput sequencing of mammalian genomes. AB - To meet the new challenge of generating the draft sequences of mammalian genomes, we describe the development of a novel high throughput 96-well method for the purification of plasmid DNA template using size-fractionated, acid-washed glass beads. Unlike most previously described approaches, the current method has been designed and optimized to facilitate the direct binding of alcohol-precipitated plasmid DNA to glass beads from alkaline lysed bacterial cells containing the insoluble cellular aggregate material. Eliminating the tedious step of separating the cleared lysate significantly simplifies the method and improves throughput and reliability. During a 4 month period of 96-capillary DNA sequencing of the Rattus norvegicus genome at the Baylor College of Medicine Human Genome Sequencing Center, the average success rate and read length derived from >1 800 000 plasmid DNA templates prepared by the direct lysis/glass bead method were 82.2% and 516 bases, respectively. The cost of this direct lysis/glass bead method in September 2001 was approximately 10 cents per clone, which is a significant cost saving in high throughput genomic sequencing efforts. PMID- 11917040 TI - Mineralocorticoid receptor malfunction: further insights from rare forms of hypertension. PMID- 11917041 TI - Controversies concerning the importance of genetic polymorphism in IgA nephropathy. PMID- 11917042 TI - Performance liquid test as a cause for sudden deaths of dialysis patients: perfluorohydrocarbon, a previously unrecognized hazard for dialysis patients. PMID- 11917043 TI - New insights in the molecular mechanisms regulating peritoneal permeability. PMID- 11917044 TI - Preparation of the dialysis patient for transplantation. PMID- 11917045 TI - CMV prophylaxis: what is valid in 2002? PMID- 11917046 TI - Cortical scintigraphy and urinary tract infection in children. PMID- 11917047 TI - Nutritional status in dialysis patients: a European consensus. AB - BACKGROUND: Malnutrition is common in dialysis patients and closely related to morbidity and mortality. Therefore, assessment of nutritional status and nutritional management of dialysis patients play a central role in everyday nephrological practice. METHODS: Achieving a consensus on key points relating to pathogenesis, clinical assessment, and nutritional management of dialysis patients. RESULTS: The assessment of nutritional status should be based on clinical assessment and biochemical parameters, including history of weight loss, per cent standard weight, body mass index, muscle mass, subcutaneous fat mass, and plasma albumin, creatinine, bicarbonate and cholesterol. Co-morbid conditions should be assessed and C-reactive protein (CRP) measured--as a marker of inflammation--as there is a close relation between malnutrition, on one side, and co-morbid conditions and inflammation on the other. For a more detailed assessment, subjective global assessment of nutritional status is a well validated tool, and dual-energy X-ray absorptiometry (DEXA) is a useful method for routine assessment of lean body mass. Anthropometric methods are also useful. They are cheap and easy to apply, although less precise than DEXA. The recommended daily protein intake is at least 1.2 g/kg standard body weight and the energy intake 35 kcal/kg standard body weight (BW), in patients <60 years, and 30 kcal/kg standard BW in patients >60 years. The standard bicarbonate level should be at least 22 mmol/l. If CRP is >10 mg/l, it is important to seek and treat the underlying cause. Adequate dialysis (for haemodialysis: Kt/V >1.2) should be ensured and, although no definite evidence of the importance of dialysis water quality is available, the opinion of the authors is that the water quality should be high. The role of the biocompatibility of the dialysis membrane is still not clear. The dietitian plays a pivotal role in the nutritional care of dialysis patients, and patients should be provided with dietary counselling from the start of substitutive treatment in order to meet the recommended nutritional intakes. Dietary counselling can also play an important role in an integrated treatment of hyperphosphataemia, although most patients will also need phosphate binders if they have an adequate protein intake. CONCLUSION: Malnutrition assessment and treatment is a great challenge for nephrological care. Achieving evidence-based consensus can help in implementing the progress of knowledge in clinical practice. PMID- 11917048 TI - Quantitative gene expression of TGF-beta1, IL-10, TNF-alpha and Fas Ligand in renal cortex and medulla. AB - BACKGROUND: Inflammatory, fibrogenic or apoptotic processes in the kidney are regulated by intra- and intercellular mediators. Intrarenal upregulation of genes may precede structural changes by days and can be examined in extremely small amounts of tissue. With the advent of new quantitative PCR methods results of gene expression are available within few hours after kidney biopsies. METHODS: In order to establish reference values for intrarenal gene expression of TGF-beta1, IL-10, TNF-alpha and Fas Ligand in renal cortex and medulla, we analysed 28 histologically normal kidney samples available after tumour nephrectomy by quantitative real-time PCR. After reverse transcription of isolated RNA, cDNA aliquots were quantified for target genes using the threshold cycle (C(t)) method normalized for the house keeping gene GAPDH. RESULTS: Expression of target genes was lower in cortex as compared to medulla, but the differences were only significant for IL-10 (P=0.0125). TGF-beta1 was found with the highest gene expression about five PCR-cycles (delta C(t)) after GAPDH with markedly lower results for TNF-alpha (delta C(t) approximately 9), IL-10 (approximately 12) and Fas Ligand (approximately 14). CONCLUSIONS: These results are the first reported reference values for human intrarenal gene expression which should facilitate interpretation of data from native or transplant kidneys in future studies. PMID- 11917049 TI - Sphingosine 1-phosphate stimulates rat mesangial cell proliferation from outside the cells. AB - BACKGROUND: Proliferation of mesangial cells (MCs) is the initial step in glomerulonephritis, and platelet-derived mediators have been shown to play a significant role in this proliferation. Sphingosine 1-phosphate (S1P), one of the sphingolipids, is abundantly stored in platelets and is released upon stimulation. We examined the effects of S1P and related sphingolipids on the cell fate of cultured MCs in order to elucidate potential roles of these lipid mediators in glomerulonephritis. METHODS: Cell proliferation was evaluated by bromodeoxy uridine (BrdU) incorporation together with MTS assay. Apoptosis of MCs was evaluated by examining annexin V staining and typical morphological changes in nuclei. We also examined the metabolism of [(3)H]sphingosine in MCs in either the presence or absence of platelet-derived growth factor (PDGF). The expression of endothelial differentiation genes (edg), which are the cell surface receptors for S1P in MCs, was examined by RT-PCR. RESULTS: S1P, but not the other sphingolipids, stimulated MC proliferation. In contrast, dimethylsphingosine (DMS) induced apoptosis in the MCs. The amount of sphingosine (Sph) converted into S1P was small and was not affected by PDGF. This observation suggested that Sph kinase activity producing S1P from Sph was low in the MCs. Furthermore, expression of edg-1, -2 and -5 in MCs was confirmed by RT-PCR. CONCLUSIONS: Our observations suggest that S1P stimulates MC proliferation from outside the cells, and not as a second messenger for PDGF. The modulation of MC fate with sphingolipids may provide possible strategies for the treatment of glomerulonephritis. PMID- 11917050 TI - Long-term renal injury in ANCA-associated vasculitis: an analysis of 31 patients with follow-up biopsies. AB - BACKGROUND: We reported previously that in renal disease in relation to antineutrophil cytoplasm auto-antibodies (ANCA)-associated vasculitis, renal outcome correlates better with the percentage of normal glomeruli than with separate active lesions. This may imply that glomeruli, once affected by necrotizing and crescentic lesions, are irreversibly damaged. We quantified and evaluated the course of renal lesions in the present study. METHODS: We retrospectively analysed 31 patients with renal disease in relation to ANCA associated vasculitis, all treated with immunosuppressive drugs. In all patients, a renal biopsy was performed at diagnosis. A follow-up biopsy was performed in all patients on the indication of a suspected renal relapse, after a mean interval of 31 months. RESULTS: The mean percentage of normal glomeruli in the renal biopsy did not change over time (29% in the initial and 30% in the follow up biopsy). The mean percentage of glomeruli with crescents, however, significantly decreased from 57 to 30% (P<0.001). The percentage of glomerulosclerosis significantly increased from 12 to 39% (P<0.001). The data were independent of diagnosis, gender, age, time interval between the biopsies, and treatment. CONCLUSIONS: This is the first study to quantify glomerular changes between two time points in patients with renal vasculitis. Our results suggest that, on average, no new glomeruli are recruited into the active disease process. The sum of the percentage of crescentic and sclerotic glomeruli in the initial biopsies is larger than the percentage of sclerotic glomeruli in the follow-up biopsies. Thus, therapy seems not only to prevent normal glomeruli from being recruited into the active disease process for a certain time, but seems also to allow part of the active lesions to revert into a normal phenotype, although another part of the active lesions will be transformed to a chronic phenotype. PMID- 11917051 TI - Randomized controlled crossover study of the effect on proteinuria and blood pressure of adding an angiotensin II receptor antagonist to an angiotensin converting enzyme inhibitor in normotensive patients with chronic renal disease and proteinuria. AB - BACKGROUND: Proteinuria and hypertension have independent deleterious effects on the progression of chronic renal disease. The objectives of this study were to determine whether the addition of Candesartan, an angiotensin II receptor antagonist, would reduce proteinuria and blood pressure in normotensive patients with chronic renal disease already receiving an angiotensin converting enzyme inhibitor (ACEI). METHODS: This was an open randomized controlled crossover study conducted in a private consultant practice in Melbourne. Sixty patients, aged 23 75, who had chronic renal disease and stable proteinuria over 0.5 g in 24 h and were receiving an ACEI, were enrolled in the study. The patients were randomized to have 8 mg of Candesartan added in the first or second of two 12-week study periods. The primary end point was urine protein excretion, which was measured every 2 weeks for the 24-week period. Secondary end points included systolic and diastolic blood pressure, serum creatinine, urea and potassium levels. Candesartan was added against a background of standard care, which included other blood pressure lowering therapy. RESULTS: Lower urine protein excretion 2.4 vs 2.0 g in 24 h (P<0.04, difference 0.45, CI 0.01, 0.9) and lower levels of systolic blood pressure 134 vs 128 mmHg (P<0.001, difference 6.4, CI 3.2, 9.6) and diastolic blood pressure 82 vs 80 mmHg (P<0.008, difference 2.7, CI 0.7, 4.6) were observed when Candesartan, 8 mg, was added to a regimen, which included an ACEI. No rise in serum creatinine occurred but there was a significant rise in urea, during the Candesartan arm of the study, from 12.3 to 13.8 mmol/l (P<0.001). The addition of 8 mg of Candesartan in normotensive patients with chronic renal disease receiving ACEI appeared safe and was not accompanied by adverse effects apart from postural hypotension in three patients and a serum potassium level of 6.3 mmol/l in one. CONCLUSIONS: In a private consulting practice setting, the addition of 8 mg of Candesartan in normotensive patients with chronic renal disease and proteinuria receiving an ACEI reduced proteinuria and blood pressure. The combination of Candesartan and ACEI appeared safe in this setting and may offer additional protection in preventing progression in chronic renal disease. Although the reduction of proteinuria was small (0.45 g/24 h) this reflected in part a lack of response in diabetic nephropathy and in part a marked rise in proteinuria after ceasing Candesartan in patients who did not complete the Candesartan arm of the study. PMID- 11917052 TI - Impaired creatinine secretion after an intravenous creatinine load is an early characteristic of the nephropathy of sickle cell anaemia. AB - BACKGROUND: The capacity to increase the tubular secretion of creatinine (TS(cr)) after an intravenous creatinine load (stimulated TS(cr)) has been found to be impaired in subclinical reduction of renal mass. We decided to investigate if this response was impaired in sickle cell anaemia (SCA) patients before there was evidence of deterioration of renal function. METHODS: Studies were done in 16 patients with homozygous SCA who had normal or supranormal glomerular filtration rate (GFR) and in 20 normal controls of similar median age (23 years). The tubular stress test (TST) consisted of 30-min clearance periods ([(125)I]iothalamate and creatinine) done before (baseline) and after (three successive post-stimulation periods) the intravenous infusion of 88.4 micromol (10 mg) of creatinine per kg of body weight. RESULTS: Baseline studies showed that the SCA patients had higher GFR and lower serum creatinine concentration. After stimulation there were no changes in GFR. In contrast, creatinine clearance increased 2.3 times in normal but not in SCA patients (P<0.001) and the TS(cr) in the first post-stimulation period was 161+/-83 nmol/kg/min in SCA patients vs 286+/-93.2 in normal controls (P<0.001). The mean TS(cr) post-stimulation was also reduced in patients with SCA (123+/-52 nmol/kg/min vs 179+/-50 in normal controls, P<0.001). Since SCA patients had lower P(cr) values, separate analysis was made of post-load clearance periods in which P(cr) was comparable in patients and in normal controls (range 177-265 micromol/l or 2-3 mg/dl) and the reduction in TS(cr) was also present in SCA patients in these study periods. CONCLUSION: Patients with SCA have impaired response to the TST before there are reductions in glomerular filtration. Therefore, a reduction in the tubular secretory reserve capacity represents an early event in the nephropathy of this condition. PMID- 11917053 TI - Smoking and proteinuria impair vasodilatory response of intrarenal arteries to nitroglycerine in patients with type 2 diabetes mellitus. AB - BACKGROUND: Few studies have addressed the effect of vasodilatory stimuli on the intrarenal arterial system in type 2 diabetes mellitus (DM), and factors affecting its responsiveness. METHODS: One hundred twenty-four patients with type 2 DM without renal failure were enrolled, and 25 subjects served as controls. Using duplex Doppler sonography, resistive indices (RI) of interlobar arteries were measured before and after sublingual nitroglycerine (NTG) (0.3 mg) spray over a 10-min period. RESULTS: Per cent changes in RI (%DeltaRI) in the DM group were significantly less than in controls (P<0.05), as was the area over the %DeltaRI-time curve (AOC-%DeltaRI, total responsiveness to nitroglycerine) (P<0.05). In the DM group, significant negative correlations were found between AOC-%DeltaRI and age (r=-0.492, P<0.0001). AOC-%DeltaRI in DM patients with proteinuria was significantly lower than without it (P<0.003). AOC-%DeltaRI in smokers was also significantly lower than in nonsmokers (P<0.05). By multiple regression analysis of the DM group, AOC-%DeltaRI was found to be significantly and independently affected by age (beta=-0.394), smoking (beta=-0.211), and the presence of proteinuria (beta=-0.270; R(2)=0.354, P<0.0001). CONCLUSIONS: Diabetic patients have a lower level of responsiveness to NTG. Advanced age, smoking, and proteinuria significantly affect response to NTG in DM patients, suggesting that advanced intrarenal arteriosclerosis may be contributory. Smoking is suggested to be a risk factor for progression of diabetic nephropathy, likely contributing to poor responsiveness of the intrarenal arterial system to vasodilatory stimuli. PMID- 11917054 TI - Radiological placement of the AshSplit haemodialysis catheter: a prospective analysis of outcome and complications. AB - BACKGROUND: The AshSplit catheter has recently been introduced as an alternative permanent tunnelled haemodialysis catheter, combining ease of insertion with good long-term patency and flow rates. METHODS: Data were collected prospectively on all the long-term tunnelled haemodialysis (AshSplit) catheters inserted radiologically between January 1998 and March 1999. Information was obtained regarding the initial insertion, ongoing catheter function and re-intervention up to September 1999. RESULTS: A total of 118 catheters were inserted in 88 patients (50 male), median (range) age 64 (20-86) years. Ultrasound guidance was used routinely and the right internal jugular vein was used in 80 (68%) cases. Initial complications occurred in 14 (11.9%) cases, which included local haemorrhage, carotid artery puncture, and air embolism. Infection occurred in 34% of catheters (2.4/1000 catheter days). Line thrombosis was documented in 20% (1.2/1000 catheter days). Satisfactory mean urea reduction ratio (URR) of 63 was obtained for all catheters. There were 47 re-interventions, mainly for fibrin sheath stripping (34) and/or thrombectomy (25). Total catheter duration was 21600 days with a 1 month cumulative survival of 87% (Kaplan-Meier probability 85%). At the end of the study, 20 (17%) catheters were still functioning, 39 (33%) had been removed electively, and 22 (18%) patients had died with a functioning catheter in situ. Catheter infection was implicated in four deaths. CONCLUSIONS: Radiological insertion of the AshSplit catheter is well tolerated, providing reliable short- and long-term dialysis access. Radiology also has a role in maintaining patency. As with all tunnelled catheters, infection remains a problem. PMID- 11917055 TI - Parathyroid hormone-independent osteoclastic resorptive bone disease: a new variant of adynamic bone disease in haemodialysis patients. AB - BACKGROUND: Osteitis fibrosa cystica (OFC) caused by secondary hyperparathyroidism is the pre-eminent form of uraemic osteodystrophy. In recent years, however, new bone abnormalities have been described. Among them adynamic bone disease (ABD) has become a focus of growing interest. Marked suppression of dynamic bone measurements with normal or near-normal static bone-forming parameters are the hallmarks of this disorder. Depressed parathyroid hormone (PTH) levels, frequently evident in this entity, have been linked causally with low bone turnover. METHODS: We reviewed bone biopsy specimens from 96 patients with end-stage renal disease undergoing chronic haemodialysis. RESULTS: We found OFC in 50% of our patients, 20% had mixed bone disease, 24% showed bone morphology of ABD and a minority (6%) had osteomalacia, mostly due to aluminium accumulation. In the patients that were affected by ABD there was a distinct subgroup with bone morphology featuring a striking increase in osteoclast number and osteoclast surface, whereas the osteoid volume, osteoid thickness, osteoblast surface, tetracycline uptake and bone formation rates were diminished as in ordinary ABD. Similarly the PTH levels in this subgroup were low or undetectable. CONCLUSION: We describe patients undergoing chronic haemodialysis with static and dynamic bone forming parameters, indistinguishable from that of ABD, but differing from the classic ABD by the presence of increased osteoclastic bone resorption. The suppressed PTH levels in this subgroup suggests that factors other than PTH activate osteoclasts in some patients on chronic haemodialysis. Uraemic cytokines and/or toxic metabolites, including beta-microglobulin, may be involved in this disorder. The precise nature of this bone abnormality remains to be defined by further studies. PMID- 11917056 TI - Total parathyroidectomy with autotransplantation in renal hyperparathyroidism: low recurrence after intra-operative tissue selection. AB - BACKGROUND: Total parathyroidectomy with simultaneous autotransplantation (AT) is a well-established surgical modality in the treatment of severe drug-resistant renal hyperparathyroidism. In literature, the high rate of graft-dependent recurrence seems a serious disadvantage. This complication can possibly be avoided by parathyroid tissue selection prior to AT. METHODS: Total parathyroidectomy with simultaneous AT was performed in 37 patients on intermittent haemodialysis treatment. Parathyroid tissue with a low proliferative potential ('A-regions') was selected for AT intra-operatively with a stereomagnifier. The mean post-operative follow-up was 37+/-24 months. RESULTS: Plasma levels of intact parathyroid hormone decreased from 1211+/-541 to 69+/-32 pg/ml, calcium from 2.49+/-0.27 to 2.17+/-0.30 mmol/l, phosphorus from 2.28+/ 0.63 to 2.11+/-0.69 mmol/l, and total alkaline phosphatases from 272+/-210 to 117+/-70 U/l. Graft-dependent recurrent hyperparathyroidism occurred in one patient after 32 months and was cured by the selective removal of five enlarged autografts. CONCLUSIONS: Simply discriminating between diffuse and nodular hyperplastic parathyroid tissue appears to be inadequate. Intra-operative tissue selection with a stereomagnifier may facilitate the identification and AT of tissue with optimal functional characteristics and a low proliferative potential, thus minimizing the rate of recurrent hyperparathyroidism. PMID- 11917057 TI - Possible involvement of increased glycoxidation and lipid peroxidation of elastin in atherogenesis in haemodialysis patients. AB - BACKGROUND: Glycoxidation and lipid peroxidation products accumulate in collagen of various tissues in haemodialysis patients with end-stage renal disease (ESRD). The purpose of this study was to test the hypothesis that increased glycoxidation and lipid peroxidation of aortic elastin is implicated in the cardiovascular complications, particularly atherosclerosis, of chronic haemodialysis patients. METHODS: Post-mortem aortic samples were obtained from 16 deceased subjects, including chronic haemodialysis patients (group 1 n=6, age 64.7+/-11.4 years) and control subjects (group 2 n=10, age 61.1+/-10.4 years). The samples were divided into three vessel wall sites: atherosclerotic intima, lesion-free intima, and media. They were sequentially treated with 0.01 M phosphate-buffered saline, collagenase, and elastase to obtain three fractions, namely soluble (SF), collagen (CF), and elastin (EF) fractions, respectively. Using spectrophotofluorometry, the pentosidine- and malondialdehyde (MDA)-linked fluorescence of these fractions was measured at wavelengths 335/385 and 390/460 (excitation/emission), respectively. RESULTS: Samples from haemodialysis patients (group 1) exhibited a significant increase in both pentosidine- and MDA-linked fluorescence of EF in atherosclerotic intima, lesion-free intima, and media samples, compared with samples from control subjects (group 2). In group 1, the levels of pentosidine- and MDA-linked fluorescence of EF were highest in atherosclerotic intima among the three aortic sites. Interestingly, in both groups, the levels of pentosidine- and MDA-linked fluorescence of EF were significantly higher than those of CF in all aortic sites. There was a strong correlation between the levels of pentosidine- and MDA-linked fluorescence in CF and EF for all aortic sites. In group 1, the pentosidine- and MDA-linked fluorescence levels of EF correlated significantly with the duration of haemodialysis in lesion-free intima and media. CONCLUSIONS: Our study provides the first biochemical evidence for a close link between aortic elastin glycoxidation and lipid peroxidation. In addition, we demonstrated high levels of these products in the aortic elastin of haemodialysis patients with ESRD. Our findings support the hypothesis that modification of aortic elastin by glycoxidation and lipid peroxidation may contribute to the development of vascular complications, particularly atherosclerosis, in patients with end-stage renal failure. PMID- 11917058 TI - Serum hepatocyte growth factor is associated with viral hepatitis, cardiovascular disease, erythropoietin treatment, and type of heparin in haemodialysis patients. AB - BACKGROUND: Increased serum hepatocyte growth factor (HGF) level is a part of the counter-system against tissue damage and predicts mortality in maintenance haemodialysis (HD) patients. We studied which of the common co-morbid and clinical conditions, and surrogates of metabolic disorders or specific organ damage determine HGF levels in these subjects. METHODS: In 86 patients, pre dialysis serum HGF, soluble endothelial markers--such as thrombomodulin (TM), von Willebrand factor and plasminogen activator inhibitor-1--and hepatitis B and C markers were measured by ELISAs. Inflammatory reactants such as C-reactive protein (CRP), alpha(1)-antitrypsin, alpha(1) acid-glycoprotein, and immunoglobulin M and G were assayed by nephelometry, and lipoprotein(a) was determined by ELISA. Cardiovascular disease (CVD) was identified on a clinical basis. RESULTS: Serum HGF was directly associated with the presence of viral hepatitis, alanine aminotransferase and TM levels, time on HD, the presence of CVD, CRP and alpha(1)-antitrypsin levels, use of unfractionated heparin (UFH) (vs enoxaparin) during HD, dose of UFH, use of recombinant erythropoietin (rHuEpo) treatment, and Kt/V. In 36 patients not treated with rHuEpo, HGF directly correlated with haemoglobin, but not with endogenous Epo levels. There was no association between HGF and the other endothelial and inflammatory markers, gender, age, smoking, cause of renal failure, body mass index, normalized protein catabolic rate, dialysate buffers, dialysers, blood pressure, antihypertensive treatment, leukocyte and platelet counts, albumin, fibrinogen, lipoprotein(a), markers of iron and calcium-phosphorus metabolism, or metabolic acidosis. Positive viral hepatitis markers, prevalent CVD and rHuEpo treatment (in descending order of significance) were independent predictors of high HGF level. In another 20 HD patients, a 4-week course of rHuEpo treatment resulted in a significant 17% increase in circulating HGF levels. CONCLUSION: Serum HGF levels in HD patients are determined by inflammatory conditions such as viral hepatitis and CVD, increase in response to rHuEpo treatment, and may be influenced by type and dose of heparin used during HD procedures. PMID- 11917059 TI - Long-term impact of renal transplantation on carotid artery properties and on ventricular hypertrophy in end-stage renal failure patients. AB - BACKGROUND: The aim of this study was to examine prospectively the impact of renal transplantation on the morphological and functional characteristics of the carotid arteries and heart in a group of end-stage renal failure patients without overt cardiovascular disease, followed up for >3 years. METHODS: Twenty-two patients were evaluated 2-3 weeks after renal transplantation, and again 12 and 40 months post-transplant, using high resolution ultrasound imaging and echocardiography. RESULTS: Kidney and patient survival were 100% at the end of follow-up without any major cardiovascular events. After 40+/-1.2 months, carotid morphological parameters were normalized: carotid intima-media thickness fell from 788+/-24 to 676+/-32 microm (P<0.01) and the carotid wall/lumen ratio fell from 118+/-3 to 103+/-3 microm (P<0.01). Significant reduction of left ventricular (LV) posterior wall thickness (11.5+/-0.2 to 11.3+/-0.2 mm, P<0.05) and LV mass index (172+/-9 to 158+/-8 g/m(2), P<0.01) was already observed after 12+/-0.2 months. Further reduction of LV posterior wall thickness (10.4+/-0.3 mm, P<0.01) and of LV mass index (136+/-7 g/m(2), P<0.01) also occurred after 40+/ 1.2 months. However, carotid distensibility (19.5+/-2.1 vs 22+/-2.4, not significant (NS)) and LV compliance (early to atrial flow ratio: 1.2+/-0.1 vs 1.3+/-0.1, NS) remained abnormal, and normalization of the LV mass was attained by only 25% of the patients with LV hypertrophy on baseline. Multiple stepwise regression analysis showed that the rate of change of reduction of the intima media thickness was influenced by age (negative association, P<0.001) and was positively related to white race (P<0.05), female sex (P<0.01) and to the parallel reduction of maximum carotid diameter (P<0.001). Reduction of LV mass index over time was negatively related to the duration of dialysis treatment and to the parallel increase observed in body mass index and haematocrit, and was positively related to the simultaneous reduction of diastolic blood pressure (P<0.01 for all variables). CONCLUSIONS: Successful renal transplantation improves but does not cause complete regression of the cardiovascular alterations of end-stage renal disease. Only intima-media thickness was normalized by transplantation, whereas LVMI and carotid and ventricular distensibility remained abnormal. The results suggest that extended duration of dialysis, weight gain, high blood pressure and high haematocrit may adversely affect the rate of change of post-transplant cardiovascular hypertrophy. PMID- 11917060 TI - The deletion polymorphism of the ACE gene is not an independent risk factor for renal scarring in children with vesico-ureteric reflux. AB - BACKGROUND: The deletion (D) polymorphism of the gene encoding angiotensin-I converting enzyme has been implicated as a risk factor for progressive renal disease in several conditions. This study was designed to evaluate the association between homozygosity for the D allele and susceptibility to renal scarring in children with vesico-ureteric reflux (VUR). METHODS: Two-hundred-and six children with VUR (all grades) were recruited into the study. Patients were stratified into two groups according to the presence or absence of renal scarring. One-hundred-and-twelve patients (group 1) had evidence of renal scarring. Ninety-four children had no evidence of renal scarring (group 2). ACE genotypes were determined by polymerase chain reaction (PCR) amplification of genomic DNA samples. RESULTS: There was no association between the DD polymorphism and the presence of renal scarring. Genotype frequencies in group 1 were: II, 29; ID, 56; and DD, 27; and in group 2 were: II, 12; ID, 52; DD, 30 (P=0.21). Neither was there evidence supporting a 'dominant' D allele. There was no association between the DD genotype and the presence of proteinuria or reduced renal function (P>0.05). Hypertension was seen more frequently in those individuals with the DD genotype, compared with the other two genotypes (P=0.012). CONCLUSION: We cannot confirm previous reports that children with vesico-ureteric reflux who are homozygous for the deletion polymorphism of the ACE gene are more susceptible to renal scarring than heterozygotes and II homozygotes. PMID- 11917061 TI - Combined therapy of tacrolimus and corticosteroids in cyclosporin-resistant or dependent idiopathic focal glomerulosclerosis: a preliminary uncontrolled study with prospective follow-up. AB - BACKGROUND: Cyclosporin has improved the outcome for steroid-resistant patients with focal glomerulosclerosis, but there is a proportion of patients that are either cyclosporin-resistant or suffer relapses, needing long-term therapy to sustain the remission. In these cases, preliminary reports suggest that tacrolimus could be an alternative therapy, but to date the evidence is limited to small series of patients with no long-term follow-up. METHODS: In this study we analysed the efficacy and safety of a combined therapy of tacrolimus and steroids in 25 patients (mean serum creatinine= 1.24+/-0.49 mg/dl; mean proteinuria=10.2+/-9.5 g/day; mean serum albumin=2.4+/-0.58 g/dl) with idiopathic primary focal glomerulosclerosis and proven resistance to or dependence on cyclosporin A. RESULTS: After a 6 months trial of tacrolimus and steroids, proteinuria decreased in 17 patients (68%) (complete remission in 10 patients (40%), partial remission in two patients (8%) and a moderate reduction in proteinuria to levels <3 g/day was seen in five additional patients (20%)). The only predictor of response to tacrolimus was a previous response to cyclosporin and prednisone, either as a complete or partial remission (remission rate 75% vs 15.3; P=0.036). Mean time to remission was 112+/-24 days. After tacrolimus discontinuation, 13/17 patients (76%) relapsed and were treated with a second trial of tacrolimus for 1 year, achieving complete remission in five patients (38.4%), partial remission in four patients (30.7%) and reduction of proteinuria <3 g/day in four patients (30.7%). After 2 years of follow-up, 12 patients (48%) were on sustained remission. The main side effect was acute reversible nephrotoxicity (40%). Predictors of renal toxicity were age (P=0.037), baseline creatinine (P=0.046) and tacrolimus trough level (P=0.001). CONCLUSIONS: We conclude that combined therapy of tacrolimus and steroids induce sustained remission of proteinuria in a significant number of patients with idiopathic focal glomerulosclerosis whose disease was not controlled by the standard therapy of steroids and cyclosporin A. PMID- 11917062 TI - Glomerulonephritis caused by Actinobacillus actinomycetemcomitans mimicking c ANCA-positive vasculitis. PMID- 11917063 TI - Simultaneous relapse of Graves' disease and minimal change glomerular disease. PMID- 11917064 TI - Nephrotic syndrome in a patient with renal amyloidosis due to polyangiitis overlap syndrome. PMID- 11917065 TI - Nephrotic syndrome in early pregnancy--is renal biopsy always necessary? PMID- 11917066 TI - Recreational drug abuse in a dialysis patient. PMID- 11917067 TI - Hypercalcaemia-induced renal failure-a mystery. PMID- 11917068 TI - An unusual presentation of renal failure. PMID- 11917069 TI - Fibrate-induced increase in serum urea and creatinine levels. PMID- 11917070 TI - The mother of an Alport's syndrome patient: a safe kidney donor? PMID- 11917071 TI - Successful treatment of factor H-related haemolytic uraemic syndrome. PMID- 11917072 TI - Renal failure associated with a specific inhibitor of BCR-ABL tyrosine kinase, STI 571. PMID- 11917073 TI - Renal artery thrombosis with acute renal failure after withdrawal of angiotensin converting enzyme inhibitor: a case report. PMID- 11917074 TI - Campylobacter fetus bacteraemia in a renal graft recipient. PMID- 11917075 TI - Evidence-based or stock market-driven nephrology? PMID- 11917078 TI - Identification and characterization of two phage-type RNA polymerase cDNAs in the moss Physcomitrella patens: implication of recent evolution of nuclear-encoded RNA polymerase of plastids in plants. AB - We isolated two cDNAs for the genes PpRPOT1 and PpRPOT2 that encode phage-type RNA polymerases (RPOTs) from Physcomitrella patens. Transcriptional activity of the encoded proteins was demonstrated by an in vitro transcription assay. Transiently expressed RPOT green fluorescent protein fusion proteins were both targeted to mitochondria. These results suggest that both PpRPOT1 and PpRPOT2 proteins function as mitochondrial RNA polymerases. Detailed phylogenetic analysis using neighbor-joining and maximum-likelihood methods with both DNA and protein sequences indicated that the two genes of P. patens form a sister group to all flowering plant genes. This suggests that the gene duplication leading to the production of plastid-type isozymes occurred after the separation of vascular plant lineage from bryophyte lineage. We therefore suggest that the generation of nuclear-encoded RNA polymerase of chloroplast is a rather recent event during the evolution of land plants. PMID- 11917079 TI - Occurrence and localization of apocarotenoids in arbuscular mycorrhizal plant roots. AB - The core structure of the yellow pigment from arbuscular mycorrhizal (AM) maize roots contains the apocarotenoids mycorradicin (an acyclic C14 polyene) and blumenol C cellobioside (a C13 cyclohexenone diglucoside). The pigment seems to be a mixture of different esterification products of these apocarotenoids. It is insoluble in water and accumulates as hydrophobic droplets in the vacuoles of root cortical cells. Screening 58 species from 36 different plant families, we detected mycorradicin in mycorrhizal roots of all Liliopsida analyzed and of a considerable number of Rosopsida, but also species were found in which mycorradicin was undetectable in mycorrhizal roots. Kinetic experiments and microscopic analyses indicate that accumulation of the yellow pigment is correlated with the concomitant degradation of arbuscules and the extensive plastid network covering these haustorium-like fungal structures. The role of the apocarotenoids in mycorrhizal roots is still unknown. The potential C40 carotenoid precursors, however, are more likely to be of functional importance in the development and functioning of arbuscules. PMID- 11917080 TI - Induced expression of a temperature-sensitive leucine-rich repeat receptor-like protein kinase gene by hypersensitive cell death and wounding in tobacco plant carrying the N resistance gene. AB - A gene encoding a receptor-like protein kinase was isolated as the gene induced in the early period of N gene-dependent hypersensitive cell death in tobacco leaves. The kinase domain expressed as a glutathione S-transferase fusion protein was capable of autophosphorylation, indicating that this gene encodes an active protein kinase. A high level of the transcript accumulated before necrotic lesion formation in tobacco mosaic virus (TMV)-inoculated tobacco leaves carrying the N gene but it was low in a tobacco cultivar lacking the N gene. A small but reproducible increase in the transcript was found 1-2 h after a temperature shift from 30 degrees C to 20 degrees C even in healthy leaves, suggesting the gene expression is temperature sensitive. The gene was named WRK for wound-induced receptor-like protein kinase, because the transcript increased to a maximum within 15-30 min of wounding. In suspension cultured tobacco cells, an increase in the transcript was found 15 min after transfer to a new medium, but it was suppressed under high osmotic pressures. The wound-induced WRK accumulation was enhanced by cycloheximide treatment, but not by known defense signal compounds (salicylic acid, jasmonic acid, 1-aminocyclopropan-1-carboxylic acid and abscisic acid) and some plant hormones. Thus, WRK is a wound-inducible and temperature sensitive protein kinase gene induced before hypersensitive cell death probably through unknown signaling pathways. PMID- 11917082 TI - Expression of the AtGH3a gene, an Arabidopsis homologue of the soybean GH3 gene, is regulated by phytochrome B. AB - Light is one of the most crucial environmental cues for plants. Phytochrome, one of the major photoreceptors of plants, regulates expression of many genes. We screened for Arabidopsis promoter trap lines that exhibited light-dependent reporter gene expression. Molecular analysis of one such line revealed that a reporter gene was inserted near an Arabidopsis homologue of the soybean GH3 gene, AtGH3a. We performed RNA gel blot analysis to further characterize the response of the AtGH3a gene to light. In response to the end-of-day far-red light treatment, the expression increased substantially. Analysis of the phyB-deficient mutant indicated that this light response is under the control of phytochrome B. The expression was also induced by exogenous auxin. Furthermore, the light response was substantially reduced in the auxin-related axr2 mutant. Taken together, it is suggested that phytochrome B regulates the expression of genes by altering the levels of auxin. PMID- 11917081 TI - Activation of the promoters of Arabidopsis genes for the branched-chain alpha keto acid dehydrogenase complex in transgenic tobacco BY-2 cells under sugar starvation. AB - Sugar starvation exerted by sub-10 mM levels of sucrose on Arabidopsis T87 suspension-cultured cells triggered marked accumulation of the transcripts of genes for E1beta and E2 subunit of the branched-chain alpha-keto acid dehydrogenase complex. Similar levels of sugar starvation increased the luciferase activity in transgenic tobacco BY-2 lines expressing the Arabidopsis E1beta- or E2-promoter-luciferase fusion gene. These results indicate that sugar levels tightly regulate the E1beta and E2 promoter activity in the heterologous plant system. We further showed in the transgenic tobacco BY-2 lines that sugar starvation-induced activation of the E1beta and E2 promoters was prevented by K 252a, an inhibitor of Ser/Thr protein kinase, and was enhanced by okadaic acid, an inhibitor of protein phosphatases. By contrast, the cauliflower mosaic virus 35S promoter activity in sugar-starved BY-2 cells was not significantly affected by K-252a and only slightly enhanced by okadaic acid. Taken together, we propose that transcriptional activation of genes for the branched-chain alpha-keto acid dehydrogenase complex and its modulation by specific protein kinases/phosphatases are of critical importance in branched-chain amino acid catabolism in plant cells under sugar starvation. PMID- 11917083 TI - Nitric oxide production mediated by nitrate reductase in the green alga Chlamydomonas reinhardtii: an alternative NO production pathway in photosynthetic organisms. AB - Biological activity of nitric oxide (NO) production was investigated in the unicellular green alga Chlamydomonas reinhardtii. An NO specific electrode detected a rapid increase in signal when nitrite (NO(2)(-)) was added into a suspension of C. reinhardtii intact cells in the dark. The addition of KCN or the NO quencher bovine hemoglobin completely abolished the signal, verifying that the nitrite-dependent increase in signal is due to enzymatic NO production. L arginine, the substrate for NO synthase, did not induce detectable NO production and the NOS inhibitor N(omega)-nitro-L-arginine showed no inhibitory effect on the nitrite-dependent production of NO. Illuminating cells showed a significant suppressive effect on NO production. When the photosynthetic electron transport inhibitor 3-(3,4-dichlorophenyl)-1,1-dimethylurea was present in the suspension, C. reinhardtii cells produced NO after the addition of nitrite even under illumination. Kinetic and microscopic observations, using the intracellular fluorescent NO probe 4,5-diaminofluorescein-2 diacetate, both demonstrated that NO was produced within the cells in response to the addition of nitrite. The Chlamydomonas mutant cc-2929, which lacks nitrate reductase (NR) activity, did not display any of the responses observed in the wild-type cells. The results presented here provide direct in vivo evidence to confirm that NR is involved in the nitrite-dependent NO production in the green alga. PMID- 11917084 TI - Formation of corymb-like inflorescences due to delay in bolting and flower development in the corymbosa2 mutant of Arabidopsis. AB - Among the wild-type ecotypes of Arabidopsis thaliana whose shape of inflorescence is categorized as raceme, the ecotype Landsberg harboring the erecta (er) mutation shows a corymb-like inflorescence, namely, a compact inflorescence with a flattened arrangement of flower buds at the tip. The fact that the ER gene encodes a receptor-like protein kinase implies the presence of a signaling cascade responsible for the inflorescence morphology of flowering plants. We report here the characterization of another mutant with a corymb-like inflorescence, named corymbosa2 (crm2), and the isolation of the CRM2 gene. While the er mutation causes a severe reduction in the length of pedicels, the crm2 mutation results in a significant delay in the initiation of internode elongation and in the development of flowers, despite having little effect on the timing of floral induction. Consequently, the number of flower buds is apparently increased at the tip of crm2 inflorescence. The crm2 er double mutant shows an additive phenotype. These results suggest that CRM2 and ER may act in different ways to generate wild-type inflorescence. The CRM2 gene was isolated by positional cloning and appears to encode a polypeptide with no significant homology to known sequences. PMID- 11917085 TI - Temporal and spatial expression pattern of the OSVP1 and OSEM genes during seed development in rice. AB - The spatial and temporal expression patterns of the rice VP1 (OSVP1) gene, as well as the OSEM gene which it controls, were studied during seed development by in situ hybridization and immuno-localization techniques. The expression of OSVP1 could be detected in embryos as early as 2-3 d after pollination (DAP) and thereafter became preferentially localized to shoot, radicle and vascular tissues during the embryo development at both the mRNA and protein levels. In the aleurone layers, OSVP1 mRNA and protein were detected after 6 DAP. OSEM mRNA was detectable after 6 DAP in the embryo and aleurone tissue. The spatial distribution within the embryo of OSEM mRNA and OSVP1 mRNA/protein was very similar after 6 DAP. Transgenic rice carrying a beta-glucuronidase (GUS) gene transcribed from a chimeric promoter consisting of the CaMV 35S minimal promoter (-46) and the 55-bp promoter fragment of OSEM, minimally required for ABA and VP1 regulation, also exhibited a spatial pattern of GUS expression similar to that of OSEM and OSVP1. These results suggest that (OS)VP1 is a major determinant not only of the seed specificity but also of the spatial pattern of OSEM expression in the developing seed. PMID- 11917086 TI - Isolation of MADS-box genes from sweet potato (Ipomoea batatas (L.) Lam.) expressed specifically in vegetative tissues. AB - New MADS-domain genes, IbMADS3 and IbMADS4, were isolated from pigmented and tuber-forming root tissue in sweet potato (Ipomoea batatas L.). Both genes were expressed preferentially in vegetative tissues, especially root tissues; white fibrous roots, pigmented roots, and developing tuberous roots. On sequence alignment, these genes fell into the STMADS group composed of SVP, STMADS11, STMADS16 and AGL24, which share high sequence similarity, similar expression patterns and similar function. Transcripts of these two genes in roots were found in the vascular cambium region. This particular expression pattern of these genes may lead to a higher proliferative potential of vegetative tissues, and may facilitate tuber initiation in sweet potato. These genes may lead to important information on the morphogenesis of vegetative structures. PMID- 11917087 TI - Involvement of the VEP1 gene in vascular strand development in Arabidopsis thaliana. AB - A dominant mutant line characterized by abnormal leaf venation pattern was isolated from a transgenic Arabidopsis plant pool that was generated with Agrobacterium culture harboring an Arabidopsis antisense cDNA library. In the mutant line, the phenotype was due to antisense suppression of a gene we named VEP1 (Vein Patterning). The predicted amino acid sequence of the gene contained a motif related to the mammalian death domain that is found in the apoptotic machinery. Reduced expression of the VEP1 gene resulted in the reduced complexity of the venation pattern of the cotyledons and foliar leaves, which was mainly due to the reduced number of the minor veins and their incomplete connection. The analysis of mutant embryos indicated that the phenotype was originated, at least in part, from a defect in the procambium patterning. In the mutant, the stem and root were thinner than those in wild type. This phenotype was associated with reduced vascular development. The promoter activity of the VEP1 gene was detected preferentially in the vascular regions. We propose that the death domain containing protein VEP1 functions as a positive element required for vascular strand development in Arabidopsis thaliana. PMID- 11917088 TI - Distribution and characterization of peroxisomes in Arabidopsis by visualization with GFP: dynamic morphology and actin-dependent movement. AB - Peroxisomes were visualized in living cells of various tissues in transgenic Arabidopsis by green fluorescent protein (GFP) through the addition of the peroxisomal targeting signal 1 (PTS1) or PTS2. The observation using confocal laser scanning microscopy revealed that the GFP fluorescence signals were detected as spherical spots in all cells of two kinds of transgenic plants. Immunoelectron microscopic analysis using antibodies against the peroxisomal marker protein, catalase, showed the presence of GFP in peroxisomes, confirming that GFP was correctly transported into peroxisomes by PTS1 or PTS2 pathways. It has been also revealed that peroxisomes are motile organelles whose movement might be caused by cytoplasmic flow. The movement of peroxisomes was more prominent in root cells than that in leaves, and divided into two categories: a relatively slow, random, vibrational movement and a rapid movement. Treatment with anti-actin and anti-tubulin drugs revealed that actin filaments involve in the rapid movement of peroxisomes. Moreover, abnormal large peroxisomes are present as clusters at the onset of germination, and these clusters disappear in a few days. Interestingly, tubular peroxisomes were also observed in the hypocotyl. These findings indicate that the shape, size, number and movement of peroxisomes in living cells are dynamic and changeable rather than uniform. PMID- 11917089 TI - Diurnal change of cucumber CPD photolyase gene (CsPHR) expression and its physiological role in growth under UV-B irradiation. AB - Complementary DNA encoding a putative cyclobutane pyrimidine dimer (CPD)-specific DNA photolyase (CPD photolyase) was isolated from cucumber leaves. The deduced amino acid sequence of the cDNA exhibited high similarity to that of the Arabidopsis CPD photolyase. Transformation with the cDNA restored the impaired photorepair activity of an Escherichia coli mutant, indicating that this cDNA encodes a functional cucumber CPD photolyase (CsPHR). The level of CsPHR transcripts estimated by quantitative RT-PCR as well as the CPD photorepair activity in the cucumber first leaves showed diurnal changes, peaking at 09 : 00 and 12 : 00, respectively. Supplemental UV-B irradiation in the middle of the light period had little effect on the growth of the first leaves, while the supplemental irradiation in the early morning or late afternoon strongly retarded the leaf growth. These results suggest that the diurnal change in CPD photorepair activity, which is presumably regulated by the transcript level of CsPHR, may play an important role in minimizing the growth inhibition due to UV-B irradiation. PMID- 11917090 TI - Identification and characterization of a gene encoding drought-inducible protein localizing in the bundle sheath cell of sugarcane. AB - We have identified a drought-inducible gene, designated as SoDip22, in sugarcane leaves. The cDNA encoded a hydrophilic protein with a calculated molecular mass of 15.9 kDa and the amino acid sequence was similar to that of ABA, stress and ripening-inducible protein from various plant species. ABA or mannitol-treatment of the detached leaves also induced SoDip22 expression. Stepwise homogenization of the stressed leaves showed that SoDip22 is localized in bundle sheath cells. These results suggest that SoDip22 functions to adapt to drought stress in the bundle sheath cell, and that the signaling pathway for the induction is, at least in a part, mediated by ABA. PMID- 11917091 TI - Function and genetics of dystrophin and dystrophin-related proteins in muscle. AB - The X-linked muscle-wasting disease Duchenne muscular dystrophy is caused by mutations in the gene encoding dystrophin. There is currently no effective treatment for the disease; however, the complex molecular pathology of this disorder is now being unravelled. Dystrophin is located at the muscle sarcolemma in a membrane-spanning protein complex that connects the cytoskeleton to the basal lamina. Mutations in many components of the dystrophin protein complex cause other forms of autosomally inherited muscular dystrophy, indicating the importance of this complex in normal muscle function. Although the precise function of dystrophin is unknown, the lack of protein causes membrane destabilization and the activation of multiple pathophysiological processes, many of which converge on alterations in intracellular calcium handling. Dystrophin is also the prototype of a family of dystrophin-related proteins, many of which are found in muscle. This family includes utrophin and alpha-dystrobrevin, which are involved in the maintenance of the neuromuscular junction architecture and in muscle homeostasis. New insights into the pathophysiology of dystrophic muscle, the identification of compensating proteins, and the discovery of new binding partners are paving the way for novel therapeutic strategies to treat this fatal muscle disease. This review discusses the role of the dystrophin complex and protein family in muscle and describes the physiological processes that are affected in Duchenne muscular dystrophy. PMID- 11917092 TI - Annexins: from structure to function. AB - Annexins are Ca2+ and phospholipid binding proteins forming an evolutionary conserved multigene family with members of the family being expressed throughout animal and plant kingdoms. Structurally, annexins are characterized by a highly alpha-helical and tightly packed protein core domain considered to represent a Ca2+-regulated membrane binding module. Many of the annexin cores have been crystallized, and their molecular structures reveal interesting features that include the architecture of the annexin-type Ca2+ binding sites and a central hydrophilic pore proposed to function as a Ca2+ channel. In addition to the conserved core, all annexins contain a second principal domain. This domain, which NH2-terminally precedes the core, is unique for a given member of the family and most likely specifies individual annexin properties in vivo. Cellular and animal knock-out models as well as dominant-negative mutants have recently been established for a number of annexins, and the effects of such manipulations are strikingly different for different members of the family. At least for some annexins, it appears that they participate in the regulation of membrane organization and membrane traffic and the regulation of ion (Ca2+) currents across membranes or Ca2+ concentrations within cells. Although annexins lack signal sequences for secretion, some members of the family have also been identified extracellularly where they can act as receptors for serum proteases on the endothelium as well as inhibitors of neutrophil migration and blood coagulation. Finally, deregulations in annexin expression and activity have been correlated with human diseases, e.g., in acute promyelocytic leukemia and the antiphospholipid antibody syndrome, and the term annexinopathies has been coined. PMID- 11917093 TI - The ubiquitin-proteasome proteolytic pathway: destruction for the sake of construction. AB - Between the 1960s and 1980s, most life scientists focused their attention on studies of nucleic acids and the translation of the coded information. Protein degradation was a neglected area, considered to be a nonspecific, dead-end process. Although it was known that proteins do turn over, the large extent and high specificity of the process, whereby distinct proteins have half-lives that range from a few minutes to several days, was not appreciated. The discovery of the lysosome by Christian de Duve did not significantly change this view, because it became clear that this organelle is involved mostly in the degradation of extracellular proteins, and their proteases cannot be substrate specific. The discovery of the complex cascade of the ubiquitin pathway revolutionized the field. It is clear now that degradation of cellular proteins is a highly complex, temporally controlled, and tightly regulated process that plays major roles in a variety of basic pathways during cell life and death as well as in health and disease. With the multitude of substrates targeted and the myriad processes involved, it is not surprising that aberrations in the pathway are implicated in the pathogenesis of many diseases, certain malignancies, and neurodegeneration among them. Degradation of a protein via the ubiquitin/proteasome pathway involves two successive steps: 1) conjugation of multiple ubiquitin moieties to the substrate and 2) degradation of the tagged protein by the downstream 26S proteasome complex. Despite intensive research, the unknown still exceeds what we currently know on intracellular protein degradation, and major key questions have remained unsolved. Among these are the modes of specific and timed recognition for the degradation of the many substrates and the mechanisms that underlie aberrations in the system that lead to pathogenesis of diseases. PMID- 11917094 TI - TRP channel proteins and signal transduction. AB - TRP channel proteins constitute a large and diverse family of proteins that are expressed in many tissues and cell types. This family was designated TRP because of a spontaneously occurring Drosophila mutant lacking TRP that responded to a continuous light with a transient receptor potential (hence TRP). In addition to responses to light, TRPs mediate responses to nerve growth factor, pheromones, olfaction, mechanical, chemical, temperature, pH, osmolarity, vasorelaxation of blood vessels, and metabolic stress. Furthermore, mutations in several members of TRP-related channel proteins are responsible for several diseases, such as several tumors and neurodegenerative disorders. TRP-related channel proteins are found in a variety of organisms, tissues, and cell types, including nonexcitable, smooth muscle, and neuronal cells. The large functional diversity of TRPs is also reflected in their diverse permeability to ions, although, in general, they are classified as nonselective cationic channels. The molecular domains that are conserved in all members of the TRP family constitute parts of the transmembrane domains and in most members also the ankyrin-like repeats at the NH2 terminal of the protein and a "TRP domain" at the COOH terminal, which is a highly conserved 25-amino acid stretch with still unknown function. All of the above features suggest that members of the TRP family are "special assignment" channels, which are recruited to diverse signaling pathways. The channels' roles and characteristics such as gating mechanism, regulation, and permeability are determined by evolution according to the specific functional requirements. PMID- 11917095 TI - Thyroid-stimulating hormone and thyroid-stimulating hormone receptor structure function relationships. AB - This review focuses on recent advances in the structure-function relationships of thyroid-stimulating hormone (TSH) and its receptor. TSH is a member of the glycoprotein hormone family constituting a subset of the cystine-knot growth factor superfamily. TSH is produced by the pituitary thyrotrophs and released to the circulation in a pulsatile manner. It stimulates thyroid functions using specific membrane TSH receptor (TSHR) that belongs to the superfamily of G protein-coupled receptors (GPCRs). New insights into the structure-function relationships of TSH permitted better understanding of the role of specific protein and carbohydrate domains in the synthesis, bioactivity, and clearance of this hormone. Recent progress in studies on TSHR as well as studies on the other GPCRs provided new clues regarding the molecular mechanisms of receptor activation. Such advances are a result of extensive site-directed mutagenesis, peptide and antibody approaches, detailed sequence analyses, and molecular modeling as well as studies on naturally occurring gain- and loss-of-function mutations. This review integrates expanding information on TSH and TSHR structure function relationships and summarizes current concepts on ligand-dependent and independent TSHR activation. Special emphasis has been placed on TSH domains involved in receptor recognition, constitutive activity of TSHR, new insights into the evolution of TSH bioactivity, and the development of high-affinity TSH analogs. Such structural, physiological, pathophysiological, evolutionary, and therapeutic implications of TSH-TSHR structure-function studies are frequently discussed in relation to concomitant progress made in studies on gonadotropins and their receptors. PMID- 11917096 TI - Molecular structure and physiological function of chloride channels. AB - Cl- channels reside both in the plasma membrane and in intracellular organelles. Their functions range from ion homeostasis to cell volume regulation, transepithelial transport, and regulation of electrical excitability. Their physiological roles are impressively illustrated by various inherited diseases and knock-out mouse models. Thus the loss of distinct Cl- channels leads to an impairment of transepithelial transport in cystic fibrosis and Bartter's syndrome, to increased muscle excitability in myotonia congenita, to reduced endosomal acidification and impaired endocytosis in Dent's disease, and to impaired extracellular acidification by osteoclasts and osteopetrosis. The disruption of several Cl- channels in mice results in blindness. Several classes of Cl- channels have not yet been identified at the molecular level. Three molecularly distinct Cl- channel families (CLC, CFTR, and ligand-gated GABA and glycine receptors) are well established. Mutagenesis and functional studies have yielded considerable insights into their structure and function. Recently, the detailed structure of bacterial CLC proteins was determined by X-ray analysis of three-dimensional crystals. Nonetheless, they are less well understood than cation channels and show remarkably different biophysical and structural properties. Other gene families (CLIC or CLCA) were also reported to encode Cl- channels but are less well characterized. This review focuses on molecularly identified Cl- channels and their physiological roles. PMID- 11917097 TI - Molecules, muscles, and machines: universal performance characteristics of motors. AB - Animal- and human-made motors vary widely in size and shape, are constructed of vastly different materials, use different mechanisms, and produce an enormous range of mass-specific power. Despite these differences, there is remarkable consistency in the maximum net force produced by broad classes of animal- and human-made motors. Motors that use force production to accomplish steady translational motion of a load (myosin, kinesin, dynein, and RNA polymerase molecules, muscle cells, whole muscles, winches, linear actuators, and rockets) have maximal force outputs that scale as the two-thirds power of mass, i.e., with cross-sectional area. Motors that use cyclical motion to generate force and are more subject to multiaxial stress and vibration have maximal force outputs that scale as a single isometric function of motor mass with mass-specific net force output averaging 57 N x kg(-1) (SD = 14). Examples of this class of motors includes flying birds, bats, and insects, swimming fish, various taxa of running animals, piston engines, electric motors, and all types of jets. Dependence of force production and stress resistance on cross-sectional area is well known, but the isometric scaling and common upper limit of mass-specific force production by cyclical motion motors has not been recognized previously and is not explained by an existing body of theory. Remarkably, this finding indicates that most of the motors used by humans and animals for transportation have a common upper limit of mass-specific net force output that is independent of materials and mechanisms. PMID- 11917098 TI - A small RNA regulates the expression of genes involved in iron metabolism in Escherichia coli. AB - A small RNA, RyhB, was found as part of a genomewide search for novel small RNAs in Escherichia coli. The RyhB 90-nt RNA down-regulates a set of iron-storage and iron-using proteins when iron is limiting; it is itself negatively regulated by the ferric uptake repressor protein, Fur (Ferric uptake regulator). RyhB RNA levels are inversely correlated with mRNA levels for the sdhCDAB operon, encoding succinate dehydrogenase, as well as five other genes previously shown to be positively regulated by Fur by an unknown mechanism. These include two other genes encoding enzymes in the tricarboxylic acid cycle, acnA and fumA, two ferritin genes, ftnA and bfr, and a gene for superoxide dismutase, sodB. Fur positive regulation of all these genes is fully reversed in an ryhB mutant. Our results explain the previously observed inability of fur mutants to grow on succinate. RyhB requires the RNA-binding protein, Hfq, for activity. Sequences within RyhB are complementary to regions within each of the target genes, suggesting that RyhB acts as an antisense RNA. In sdhCDAB, the complementary region is at the end of the first gene of the sdhCDAB operon; full-length sdhCDAB message disappears and a truncated message, equivalent in size to the region upstream of the complementarity, is detected when RyhB is expressed. RyhB provides a mechanism for the cell to down-regulate iron-storage proteins and nonessential iron-containing proteins when iron is limiting, thus modulating intracellular iron usage to supplement mechanisms for iron uptake directly regulated by Fur. PMID- 11917099 TI - Paranodal junction formation and spermatogenesis require sulfoglycolipids. AB - Mammalian sulfoglycolipids comprise two major members, sulfatide (HSO3-3 galactosylceramide) and seminolipid (HSO3-3-monogalactosylalkylacylglycerol). Sulfatide is a major lipid component of the myelin sheath and serves as the epitope for the well known oligodendrocyte-marker antibody O4. Seminolipid is synthesized in spermatocytes and maintained in the subsequent germ cell stages. Both sulfoglycolipids can be synthesized in vitro by using the isolated cerebroside sulfotransferase. To investigate the physiological role of sulfoglycolipids and to determine whether sulfatide and seminolipid are biosynthesized in vivo by a single sulfotransferase, Cst-null mice were generated by gene targeting. Cst(-/-) mice lacked sulfatide in brain and seminolipid in testis, proving that a single gene copy is responsible for their biosynthesis. Cst(-/-) mice were born healthy, but began to display hindlimb weakness by 6 weeks of age and subsequently showed a pronounced tremor and progressive ataxia. Although compact myelin was preserved, Cst(-/-) mice displayed abnormalities in paranodal junctions. On the other hand, Cst(-/-) males were sterile because of a block in spermatogenesis before the first meiotic division, whereas females were able to breed. These data show a critical role for sulfoglycolipids in myelin function and spermatogenesis. PMID- 11917101 TI - Rate heterogeneity among lineages of tracheophytes: integration of molecular and fossil data and evidence for molecular living fossils. AB - Many efforts to date evolutionary divergences by using a molecular clock have yielded age estimates that are grossly inconsistent with the paleontological evidence. Such discrepancies often are attributed to the inadequacy of the fossil record, but many potential sources of error can affect molecular-based estimates. In this study, we minimize the potential error caused by inaccurate topology and uncertain calibration times by using a well-supported tree, multiple genes, and multiple well-substantiated dates to explore the correspondence between the fossil record and molecular-based age estimates for major clades of tracheophytes. Age estimates varied because of gene effects, codon position, lineage effects, method of inferring branch lengths, and whether or not rate constancy was assumed. However, even methods designed to ameliorate the effects of rate heterogeneity among lineages could not accommodate the substantially slower rates observed in Marattia + Angiopteris and in the tree ferns. Both of these clades of ferns have undergone dramatic decelerations in their rates of molecular evolution and are "molecular living fossils," consistent with their relative morphological stasis for the past 165-200 million years. Similar discrepancies between the fossil record and molecular-based age estimates noted in other studies may also be explained in part by violations of rate constancy among lineages. PMID- 11917102 TI - Examination of hypotheses for the Permo-Triassic boundary extinction by carbon cycle modeling. AB - The biological extinction that occurred at the Permian-Triassic boundary represents the most extensive loss of species of any known event of the past 550 million years. There have been a wide variety of explanations offered for this extinction. In the present paper, a number of the more popular recent hypotheses are evaluated in terms of predictions that they make, or that they imply, concerning the global carbon cycle. For this purpose, a mass balance model is used that calculates atmospheric CO2 and oceanic delta13C as a function of time. Hypotheses considered include: (i) the release of massive amounts of CO2 from the ocean to the atmosphere resulting in mass poisoning; (ii) the release of large amounts of CO2 from volcanic degassing; (iii) the release of methane stored in methane hydrates; (iv) the decomposition and oxidation of dead organisms to CO2 after sudden mass mortality; and (v) the long-term reorganization of the global carbon cycle. The modeling indicates that measured short-term changes in delta13C at the boundary are best explained by methane release with mass mortality and volcanic degassing contributing in secondary roles. None of the processes result in excessively high levels of atmospheric CO2 if they occurred on time scales of more than about 1,000 years. The idea of poisoning by high levels of atmospheric CO2 depends on the absence of subthermocline calcium carbonate deposition during the latest Permian. The most far-reaching effect was found to be reorganization of the carbon cycle with major sedimentary burial of organic matter shifting from the land to the sea, resulting in less burial overall, decreased atmospheric O2, and higher atmospheric CO2 for the entire Triassic Period. PMID- 11917100 TI - Endothelial cells derived from human embryonic stem cells. AB - Human embryonic stem cells have the potential to differentiate into various cell types and, thus, may be useful as a source of cells for transplantation or tissue engineering. We describe here the differentiation steps of human embryonic stem cells into endothelial cells forming vascular-like structures. The human embryonic-derived endothelial cells were isolated by using platelet endothelial cell-adhesion molecule-1 (PECAM1) antibodies, their behavior was characterized in vitro and in vivo, and their potential in tissue engineering was examined. We show that the isolated embryonic PECAM1+ cells, grown in culture, display characteristics similar to vessel endothelium. The cells express endothelial cell markers in a pattern similar to human umbilical vein endothelial cells, their junctions are correctly organized, and they have high metabolism of acetylated low-density lipoprotein. In addition, the cells are able to differentiate and form tube-like structures when cultured on matrigel. In vivo, when transplanted into SCID mice, the cells appeared to form microvessels containing mouse blood cells. With further studies, these cells could provide a source of human endothelial cells that could be beneficial for potential applications such as engineering new blood vessels, endothelial cell transplantation into the heart for myocardial regeneration, and induction of angiogenesis for treatment of regional ischemia. PMID- 11917103 TI - Vav3 is regulated during the cell cycle and effects cell division. AB - Vav3 is a member of the family of guanine nucleotide exchange factors implicated in the regulation of Rho GTPases. Although the exact in vivo function of Vav3 is unknown, evidence from several studies indicates a role distinct from Vav2 or Vav1. Here we report that the expression of Vav3 is regulated during the cell cycle. Strikingly, Vav3 was transiently up-regulated in HeLa cells during mitosis, whereas enforced expression of Vav3 perturbed cytokinesis and led to the appearance of multinucleated cells. These effects of Vav3 were RhoA-dependent, required phosphorylation of the regulatory tyrosine 173, but were not enhanced by N-terminal truncations. Thus, this report establishes that expression of Vav3 is strictly regulated in a cell cycle-dependent manner and implicates Vav3 in the control of cytokinesis. PMID- 11917104 TI - In vitro cartilage formation by human adult stem cells from bone marrow stroma defines the sequence of cellular and molecular events during chondrogenesis. AB - One approach to resolving the complexities of chondrogenesis is to examine simplified systems in vitro. We analyzed cartilage differentiation by human adult stem cells from bone marrow stroma. Marrow stromal cells were cultured as micromass pellets for 21 days in serum-free medium containing transforming growth factor (TGF)-beta3, dexamethasone, and bone morphogenetic protein (BMP)-6. Assays for pulse-labeled [3H]DNA and for total DNA indicated that there was little proliferation and a progressive loss of cells in the pellets. There were continuous increases in mRNAs for cartilage matrix (proteoglycans and COL2, -9, 10, and -11), receptors [fibroblast growth factor 2 (FGFR2) and parathyroid hormone-related peptide receptor (PTHrP-R)], and transcription factors (SOX5, -6, and -9) as demonstrated by histochemical and microarray assays. Reverse transcription-PCR assays for 11 mRNAs confirmed the microarray data. SOX4, vascular endothelial growth factor (VEGF), and matrix metalloproteinase 14 (MMP14) increased at day 1 and decreased thereafter, suggesting roles early in chondrogenesis. Also, forkhead, CD10, and MMP13 increased up to day 7 and decreased thereafter, suggesting roles in an intermediate stage of chondrogenesis. In addition, two collagens (COL3A1 and COL16A1), a signaling molecule (WNT11), a homeobox homolog (BAPX1), a receptor (IL-1R1), an IGFs modulator (IGFBP5), and a mettaloproteinase (MMP16) increased progressively up to about day 14, suggesting roles later in chondrogenesis. Our results indicate that the simplicity of the system makes it possible to define in detail the cellular and molecular events during chondrogenesis. PMID- 11917105 TI - Reversal of motor impairments in parkinsonian rats by continuous intrastriatal delivery of L-dopa using rAAV-mediated gene transfer. AB - Intrastriatal delivery of the tyrosine hydroxylase gene by viral vectors is being explored as a tool for local delivery of L-dopa in animals with lesions of the nigrostriatal pathway. The functional effects reported using this approach have been disappointing, probably because the striatal L-dopa levels attained have been too low. In the present study, we have defined a critical threshold level of L-dopa, 1.5 pmol/mg of tissue, that has to be reached to induce any significant functional effects. Using new generation high-titer recombinant adeno-associated virus vectors, we show that levels of striatal L-dopa production exceeding this threshold can be obtained provided that tyrosine hydroxylase is coexpressed with the cofactor synthetic enzyme, GTP-cyclohydrolase-1. After striatal transduction with this combination of vectors, substantial functional improvement in both drug induced and spontaneous behavior was observed in rats with either complete or partial 6-hydroxydopamine lesions of the nigrostriatal pathway. However, complete reversal of motor deficits occurred only in animals in which part of the striatal dopamine innervation was left intact. Spared nigrostriatal fibers thus may convert L-dopa to dopamine and store and release dopamine in a more physiologically relevant manner in the denervated striatum to mediate better striatal output-dependent motor function. We conclude that intrastriatal L-dopa delivery may be a viable strategy for treatment and control of adverse side effects associated with oral L-dopa therapy such as on-off fluctuations and drug induced dyskinesias in patients with Parkinson's disease. PMID- 11917106 TI - Identification of a protein essential for a major pathway used by human cells to avoid UV- induced DNA damage. AB - When DNA replication stalls at a fork-blocking lesion, cells use damage tolerance pathways to continue replication. One pathway, "translesion synthesis," involves specialized DNA polymerases that can use damaged DNA as a template. Translesion synthesis can result in mutations (i.e., can be error-prone), but it can also be error-free. An alternative pathway has been hypothesized (sometimes called "damage avoidance"), by which cells make temporary use of an undamaged copy of the blocked sequence as a template, i.e., the newly synthesized daughter strand of the sister duplex or the allelic copy. This pathway is error-free. Evidence of the use of the daughter strand of the sister duplex as a template in intact mammalian cells has not been available heretofore. To determine whether hMms2, a ubiquitin-conjugating enzyme-like protein, plays a critical role in such damage avoidance, a human fibroblast cell strain in which both error-prone translesion synthesis and error-free damage avoidance can be detected and quantified simultaneously, and several derivative strains in which expression of hMms2 protein had been eliminated or greatly decreased, were compared for their ability to avoid translesion synthesis past UV(254nm)-induced DNA photoproducts. Loss of hMms2 protein eliminated the ability of the latter strains to use an allelic copy of a target gene for damage avoidance, i.e., to produce a wild-type gene from two nonfunctional allelic copies of that gene. Molecular analysis of the wild-type gene showed that this process involves gene conversion unassociated with crossing over. That the loss of hMms2 also eliminated use of the daughter strand of the sister duplex as a template for damage avoidance could be inferred from the fact that the frequency of mutations induced by UV in the single copy HPRT gene of the derivative strains was significantly higher than that observed in the parental strain. These data indicate that hMMS2 is essential for human cells to carry out damage avoidance by using either type of homolog, and that damage avoidance and translesion synthesis are alternative pathways for tolerating fork-blocking photoproducts. PMID- 11917107 TI - Stimulation of bone formation and prevention of bone loss by prostaglandin E EP4 receptor activation. AB - Bone remodeling, comprising resorption of existing bone and de novo bone formation, is required for the maintenance of a constant bone mass. Prostaglandin (PG)E2 promotes both bone resorption and bone formation. By infusing PGE2 to mice lacking each of four PGE receptor (EP) subtypes, we have identified EP4 as the receptor that mediates bone formation in response to this agent. Consistently, bone formation was induced in wild-type mice by infusion of an EP4-selective agonist and not agonists specific for other EP subtypes. In culture of bone marrow cells from wild-type mice, PGE2 induced expression of core-binding factor alpha1 (Runx2/Cbfa1) and enhanced formation of mineralized nodules, both of which were absent in the culture of cells from EP4-deficient mice. Furthermore, administration of the EP4 agonist restored bone mass and strength normally lost in rats subjected to ovariectomy or immobilization. Histomorphometric analysis revealed that the EP4 agonist induced significant increases in the volume of cancellous bone, osteoid formation, and the number of osteoblasts in the affected bone of immobilized rats, indicating that activation of EP4 induces de novo bone formation. In addition, osteoclasts were found on the increased bone surface at a density comparable to that found in the bone of control animals. These results suggest that activation of EP4 induces bone remodeling in vivo and that EP4 selective drugs may be beneficial in humans with osteoporosis. PMID- 11917108 TI - Genome sequence and comparative microarray analysis of serotype M18 group A Streptococcus strains associated with acute rheumatic fever outbreaks. AB - Acute rheumatic fever (ARF), a sequelae of group A Streptococcus (GAS) infection, is the most common cause of preventable childhood heart disease worldwide. The molecular basis of ARF and the subsequent rheumatic heart disease are poorly understood. Serotype M18 GAS strains have been associated for decades with ARF outbreaks in the U.S. As a first step toward gaining new insight into ARF pathogenesis, we sequenced the genome of strain MGAS8232, a serotype M18 organism isolated from a patient with ARF. The genome is a circular chromosome of 1,895,017 bp, and it shares 1.7 Mb of closely related genetic material with strain SF370 (a sequenced serotype M1 strain). Strain MGAS8232 has 178 ORFs absent in SF370. Phages, phage-like elements, and insertion sequences are the major sources of variation between the genomes. The genomes of strain MGAS8232 and SF370 encode many of the same proven or putative virulence factors. Importantly, strain MGAS8232 has genes encoding many additional secreted proteins involved in human-GAS interactions, including streptococcal pyrogenic exotoxin A (scarlet fever toxin) and two uncharacterized pyrogenic exotoxin homologues, all phage-associated. DNA microarray analysis of 36 serotype M18 strains from diverse localities showed that most regions of variation were phages or phage-like elements. Two epidemics of ARF occurring 12 years apart in Salt Lake City, UT, were caused by serotype M18 strains that were genetically identical, or nearly so. Our analysis provides a critical foundation for accelerated research into ARF pathogenesis and a molecular framework to study the plasticity of GAS genomes. PMID- 11917109 TI - Circadian 5-HT production regulated by adrenergic signaling. AB - Using on-line microdialysis, we have characterized in vivo dynamics of pineal 5 hydroxytryptamine (5-HT; serotonin) release. Daily pineal 5-HT output is triphasic: (i) 5-HT levels are constant and high during the day; (ii) early in the night, there is a novel sharp rise in 5-HT synthesis and release, which precedes the nocturnal rise in melatonin synthesis; and (iii) late in the night, levels are low. This triphasic 5-HT production persists in constant darkness and is influenced strongly by intrusion of light at night. We demonstrate that both diurnal 5-HT synthesis and 5-HT release are activated by sympathetic innervation from the superior cervical ganglion and show that these processes are controlled by distinct receptors. The increase in 5-HT synthesis is controlled by beta adrenergic receptors, whereas the increase in 5-HT release is mediated by alpha adrenergic signaling. On the other hand, the marked decrease in 5-HT content and release late at night is a passive process, influenced by the extent of melatonin synthesis. In the absence of melatonin synthesis, the late-night decline in 5-HT release is prevented, reaching levels roughly twice as high as that of the day value. In summary, our results demonstrate that 5-HT levels display marked circadian rhythms that depend on adrenergic signaling. PMID- 11917112 TI - Anion recognition as a method for templating pseudorotaxane formation. AB - Two thread-shaped cations, pyridinium nicotinamide and imidazolium, as their chloride and hexafluorophosphate salts, were studied with regards to complexation with hydrogen-bond-donating acyclic and macrocyclic ligands. In the latter case, the cations form pseudorotaxanes templated by the chloride anion but not hexafluorophosphate. This formation is a function of the coupling of ion-pairing between the cation and chloride anion and subsequent recognition of the anion by the macrocyclic diamide, which provides the driving force for interpenetration. We propose that this anion template principle is a general method for the construction of pseudorotaxanes and could be applied to other cationic threads, anions, and macrocyclic species. PMID- 11917110 TI - Naive T cells proliferate strongly in neonatal mice in response to self peptide/self-MHC complexes. AB - Adult naive T cells, which are at rest in normal conditions, proliferate strongly when transferred to lymphopenic hosts. In neonates, the first mature thymocytes to migrate to the periphery reach a compartment devoid of preexisting T cells. We have extensively analyzed the proliferation rate and phenotype of peripheral T cells from normal C57BL/6 and T cell antigen receptor transgenic mice as a function of age. We show that, like adult naive T cells transferred to lymphopenic mice, neonatal naive T cells proliferate strongly. By using bone marrow transfer and thymic-graft models, we demonstrate that the proliferation of the first thymic emigrants reaching the periphery requires T cell antigen receptor-self-peptide/self-MHC interactions and is regulated by the size of the peripheral T cell pool. PMID- 11917113 TI - Discrete and polymeric self-assembled dendrimers: hydrogen bond-mediated assembly with high stability and high fidelity. AB - Hydrogen bond-mediated self-assembly is a powerful strategy for creating nanoscale structures. However, little is known about the fidelity of assembly processes that must occur when similar and potentially competing hydrogen-bonding motifs are present. Furthermore, there is a continuing need for new modules and strategies that can amplify the relatively weak strength of a hydrogen bond to give more stable assemblies. Herein we report quantitative complexation studies on a ureidodeazapterin-based module revealing an unprecedented stability for dimers of its self-complementary acceptor-acceptor-donor-donor (AADD) array. Linking two such units together with a semirigid spacer that carries a first-, second-, or third-generation Frechet-type dendron affords a ditopic structure programmed to self assemble. The specific structure that is formed depends both on the size of the dendron and the solvent, but all of the assemblies have exceptionally high stability. The largest discrete nanoscale assembly is a hexamer with a molecular mass of about 17.8 kDa. It is stabilized by 30 hydrogen bonds, including six AADD x DDAA contacts. The hexamer forms and is indefinitely stable in the presence of a hexamer containing six ADD x DAA hydrogen-bonding arrays. PMID- 11917114 TI - Virus-assisted loading of polymer nanocontainer. AB - We present a DNA-containing polymeric nanocontainer using the self-assembled superstructure of amphiphilic block copolymers in aqueous solutions. To demonstrate that DNA translocation is possible across a completely synthetic block copolymer membrane, we have used a phage transfection strategy as a DNA transfer model system. For this purpose the bacterial channel forming protein LamB was reconstituted in ABA-triblock copolymer vesicles. The outer membrane protein LamB is a specific transporter for maltodextrins but also serves as a receptor for lambda phage to trigger the ejection of lambda phage DNA. We demonstrate that the functionality of the LamB protein is fully preserved despite the artificial surrounding. This leads to a type of polymeric vehicle for DNA that could be useful for gene therapy. PMID- 11917115 TI - Inclusion of methylviologen in cucurbit[7]uril. AB - The inclusion behavior of methylviologen (N,N'-dimethyl-4,4'-bipyridinium, MV) dication in cucurbit[7]uril (CB[7]) has been studied by using various spectroscopic and electrochemical methods. The inclusion complex of MV dication in CB[7] is stable thermodynamically and kinetically. The electrochemical study reveals that unlike beta-cyclodextrin, CB[7] prefers the charged species, MV dication (MV(2+)), and cation radical (MV(+)*) to the fully reduced neutral (MV(0) species as guests. Dimerization of MV(+)* is suppressed effectively by forming a stable complex with CB[7] in aqueous solution as confirmed by spectroelectrochemical experiments. Furthermore, the first redox process (MV(2+)/MV(+)*) of the MV(2+)-CB[7] complex occurs predominantly via the direct electron transfer pathway, whereas the second redox process (MV(+)*/MV(0)) occurs via both the direct and indirect pathway because of the low affinity of the fully reduced species MV(0) to CB[7]. PMID- 11917116 TI - Suppression of genome instability by redundant S-phase checkpoint pathways in Saccharomyces cerevisiae. AB - Cancer cells show increased genome rearrangements, although it is unclear what defects cause these rearrangements. Previous studies have implicated the Saccharomyces cerevisiae replication checkpoint in the suppression of spontaneous genome rearrangements. In the present study, low doses of methyl methane sulfonate that activate the intra-S checkpoint but not the G1 or G2 DNA damage checkpoints were found to cause increased accumulation of genome rearrangements in both wild-type strains and to an even greater extent in strains containing mutations causing defects in the intra-S checkpoint. The rearrangements were primarily translocations or events resulting in deletion of a portion of a chromosome arm along with the addition of a new telomere. Combinations of mutations causing individual defects in the RAD24 or SGS1 branches of the intra-S checkpoint or the replication checkpoint showed synergistic interactions with regard to the spontaneous genome instability rate. PDS1 and the RAD50-MRE11-XRS2 complex were found to be important members of all the S-phase checkpoints in suppressing genome instability, whereas RAD53 only seemed to play a role in the intra-S checkpoints. Combinations of mutations that seem to result in inactivation of the S-phase checkpoints and critical effectors resulted in as much as 12,000-14,000-fold increases in the genome instability rate. These data support the view that spontaneous genome rearrangements result from DNA replication errors and indicate that there is a high degree of redundancy among the checkpoints that act in S phase to suppress such genome instability. PMID- 11917117 TI - A physiologic signaling role for the gamma -secretase-derived intracellular fragment of APP. AB - Presenilins mediate an unusual intramembranous proteolytic activity known as gamma-secretase, two substrates of which are the Notch receptor (Notch) and the beta-amyloid precursor protein (APP). Gamma-secretase-mediated cleavage of APP, like that of Notch, yields an intracellular fragment [APP intracellular domain (AICD)] that forms a transcriptively active complex. We now demonstrate a functional role for AICD in regulating phosphoinositide-mediated calcium signaling. Genetic ablation of the presenilins or pharmacological inhibition of gamma-secretase activity (and thereby AICD production) attenuated calcium signaling in a dose-dependent and reversible manner through a mechanism involving the modulation of endoplasmic reticulum calcium stores. Cells lacking APP (and hence AICD) exhibited similar calcium signaling deficits, and-notably-these disturbances could be reversed by transfection with APP constructs containing an intact AICD, but not by constructs lacking this domain. Our findings indicate that the AICD regulates phosphoinositide-mediated calcium signaling through a gamma-secretase-dependent signaling pathway, suggesting that the intramembranous proteolysis of APP may play a signaling role analogous to that of Notch. PMID- 11917118 TI - Early expression of triggering receptors and regulatory role of 2B4 in human natural killer cell precursors undergoing in vitro differentiation. AB - In this study we analyzed the progression of cell surface receptor expression during the in vitro-induced human natural killer (NK) cell maturation from CD34(+) Lin(-) cell precursors. NKp46 and NKp30, two major triggering receptors that play a central role in natural cytotoxicity, were expressed before the HLA class I-specific inhibitory receptors. Moreover, their appearance at the cell surface correlated with the acquisition of cytolytic activity by developing NK cells. Although the early expression of triggering receptors may provide activating signals required for inducing further cell differentiation, it may also affect the self-tolerance of developing NK cells. Our data show that a fail safe mechanism preventing killing of normal autologous cells may be provided by the 2B4 surface molecule, which, at early stages of NK cell differentiation, functions as an inhibitory rather than as an activating receptor. PMID- 11917119 TI - Desensitization of mu-opioid receptor-evoked potassium currents: initiation at the receptor, expression at the effector. AB - Many G protein-coupled receptor-mediated responses desensitize within minutes. Sustained stimulation of mu-opioid receptors (MORs), which primarily signal through G(i/o) proteins, leads to activation and subsequent desensitization of G protein-coupled inwardly rectifying potassium (GIRK) currents. We observed that in neurons of the locus coeruleus, which express among the highest levels of MORs in the brain, the degree of desensitization depended on the intensity of receptor stimulation, indicating that the process is initiated at the receptor. Interestingly, while GIRK-mediated postsynaptic inhibition substantially desensitized within 15 min, presynaptic inhibition of afferent transmission, which involves other effector systems, remained constant, suggesting that the postsynaptic desensitization we observed is expressed at the effector. We show that desensitized GIRK currents can gradually be reactivated by additional G protein signals of increasing intensity and present evidence that desensitization is a G protein-mediated process. Finally, desensitization of MOR-induced GIRK currents had heterologous effects on responses mediated by other G protein coupled receptors converging onto the same population of GIRK channels. Taken together, our results provide evidence for a form of desensitization mediated by a slowly developing G protein-dependent pathway, initiated at the MORs and leading to competitive inhibition of GIRK channel activation. This implies that MORs exert a bidirectional action on GIRK channels. PMID- 11917120 TI - Aquaporin 1 regulates GTP-induced rapid gating of water in secretory vesicles. AB - The swelling of secretory vesicles has been implicated in exocytosis, but the underlying mechanism of vesicle swelling remains largely unknown. Zymogen granules (ZGs), the membrane-bound secretory vesicles in exocrine pancreas, swell in response to GTP mediated by a G(alpha)i3 protein. Evidence is presented here that the water channel aquaporin-1 (AQP1) is present in the ZG membrane and participates in rapid GTP-induced vesicular water gating and swelling. Isolated ZGs exhibit low basal water permeability. However, exposure of granules to GTP results in a marked potentiation of water entry. Treatment of ZGs with the known water channel inhibitor Hg2+ is accompanied by a reversible loss in both the basal and GTP-stimulatable water entry and vesicle swelling. Introduction of AQP1 specific antibody raised against the carboxyl-terminal domain of AQP1 blocks GTP stimulable swelling of vesicles. Our results demonstrate that AQP1 associated at the ZG membrane is involved in basal as well as GTP-induced rapid gating of water in ZGs of the exocrine pancreas. PMID- 11917121 TI - Mutation of melanosome protein RAB38 in chocolate mice. AB - Mutations of genes needed for melanocyte function can result in oculocutaneous albinism. Examination of similarities in human gene expression patterns by using microarray analysis reveals that RAB38, a small GTP binding protein, demonstrates a similar expression profile to melanocytic genes. Comparative genomic analysis localizes human RAB38 to the mouse chocolate (cht) locus. A G146T mutation occurs in the conserved GTP binding domain of RAB38 in cht mice. Rab38(cht)/Rab38(cht) mice exhibit a brown coat similar in color to mice with a mutation in tyrosinase related protein 1 (Tyrp1), a mouse model for oculocutaneous albinism. The targeting of TYRP1 protein to the melanosome is impaired in Rab38(cht)/Rab38(cht) melanocytes. These observations, and the fact that green fluorescent protein tagged RAB38 colocalizes with end-stage melanosomes in wild-type melanocytes, suggest that RAB38 plays a role in the sorting of TYRP1. This study demonstrates the utility of expression profile analysis to identify mammalian disease genes. PMID- 11917122 TI - Lineage infidelity in myeloid cells with TCR gene rearrangement: a latent developmental potential of proT cells revealed by ectopic cytokine receptor signaling. AB - The most immature lymphoid-committed progenitors in both the bone marrow (common lymphoid progenitor) and thymus (proT1) maintain a latent granulocyte/macrophage (G/M) differentiation potential that can be initiated by signals emanating from exogenously expressed IL-2 receptors. In this study, we investigate at which developmental stage thymocytes lose this G/M differentiation potential. We demonstrate that the next maturational stage after proT1 cells (proT2), but not preT (TN3) cells, can convert cell fate from lymphoid to myeloid in response to ectopic IL-2 receptor signaling in human IL-2Rbeta transgenic mice. It is significant that approximately 10% of clonogenic G/M colonies derived from proT cells of IL-2Rbeta transgenic mice have DJ rearrangement specifically at the Dbeta1 but not Dbeta2 segment in the TCRbeta locus. No TCR gene rearrangement is observed in G/M cells from nontransgenic mice, suggesting that the G/M cells we observe in this system were truly lymphoid-committed before stimulation with IL 2. In addition, Dbeta1 and Dbeta2 DJ rearrangement of the TCRbeta gene may be differentially regulated and thus serve as markers for distinct proT cell maturational stages. PMID- 11917123 TI - The procaspase-8 isoform, procaspase-8L, recruited to the BAP31 complex at the endoplasmic reticulum. AB - BAP31 is an integral protein of the endoplasmic reticulum membrane and a substrate of caspase-8. Here, we describe the procaspase-8 isoform, procaspase 8L, which is ubiquitously expressed and selectively recruited to the BAP31 complex in response to apoptotic signaling by E1A. Procaspase-8L is characterized by the N-terminal extension (Nex) domain, which extends procaspase-8/a at the N terminus and is required for selective association of procaspase-8L with the BAP31 complex. Gene deletion identified BAP31 and related BAP29 as required for processing of procaspase-8L in response to E1A, by a FADD-independent mechanism that was blocked by BCL-2. Further, Bap29,31 deletion, as well as a Nex-domain dominant-negative mutant, curtailed the activation of downstream caspases (IETDase and DEVDase) and cell death in response to E1A. Preferential recruitment of procaspase-8L by the BAP31 complex at the endoplasmic reticulum suggests an additional pathway for regulating initiator caspase-8 during apoptosis. PMID- 11917124 TI - Leukotriene A4 hydrolase: selective abrogation of leukotriene B4 formation by mutation of aspartic acid 375. AB - Leukotriene A4 (LTA4, 5S-trans-5,6-oxido-7,9-trans-11,14-cis-eicosatetraenoic acid) hydrolase (LTA4H)/aminopeptidase is a bifunctional zinc metalloenzyme that catalyzes the final and rate-limiting step in the biosynthesis of leukotriene B4 (LTB4, 5S,12R-dihydroxy-6,14-cis-8,10-trans-eicosatetraenoic acid), a classical chemoattractant and immune modulating lipid mediator. Two chemical features are key to the bioactivity of LTB4, namely, the chirality of the 12R-hydroxyl group and the cis-trans-trans geometry of the conjugated triene structure. From the crystal structure of LTA4H, a hydrophilic patch composed of Gln-134, Tyr-267, and Asp-375 was identified in a narrow and otherwise hydrophobic pocket, believed to bind LTA4. In addition, Asp-375 belongs to peptide K21, a previously characterized 21-residue active site-peptide to which LTA4 binds during suicide inactivation. In the present report we used site-directed mutagenesis and x-ray crystallography to show that Asp-375, but none of the other candidate residues, is specifically required for the epoxide hydrolase activity of LTA4H. Thus, mutation of Asp-375 leads to a selective loss of the enzyme's ability to generate LTB4 whereas the aminopeptidase activity is preserved. We propose that Asp-375, possibly assisted by Gln-134, acts as a critical determinant for the stereoselective introduction of the 12R-hydroxyl group and thus the biological activity of LTB4. PMID- 11917125 TI - Human receptors for sweet and umami taste. AB - The three members of the T1R class of taste-specific G protein-coupled receptors have been hypothesized to function in combination as heterodimeric sweet taste receptors. Here we show that human T1R2/T1R3 recognizes diverse natural and synthetic sweeteners. In contrast, human T1R1/T1R3 responds to the umami taste stimulus l-glutamate, and this response is enhanced by 5'-ribonucleotides, a hallmark of umami taste. The ligand specificities of rat T1R2/T1R3 and T1R1/T1R3 correspond to those of their human counterparts. These findings implicate the T1Rs in umami taste and suggest that sweet and umami taste receptors share a common subunit. PMID- 11917126 TI - Function of quaking in myelination: regulation of alternative splicing. AB - Proteomic diversity is frequently achieved by alternative RNA-splicing events that can be fine-tuned in tissue-specific and developmentally regulated ways. Understanding this type of genetic regulation is compelling because of the extensive complexity of alternative splicing found in the nervous system. quaking (qk), one of the classical mouse dysmyelination mutants, is defective for the expression of myelin-associated glycoprotein (MAG), and the misregulation of MAG pre-mRNA alternative splicing is implicated as a causal factor. The qk locus encodes several RNA-binding proteins with heterogeneous nuclear ribonucleoprotein K-type homology, a characteristic of several known alternative splicing regulators. Here we test the nuclear-localized qk isoform (QKI-5) for its ability to regulate alternative splicing of MAG pre-mRNA in transient coexpression assays. QKI-5 exhibits properties of a negative regulator of MAG exon 12 alternative splicing. An intronic sequence element required for the repressive function and binding of QKI-5 is also identified. Direct evidence for irregularities in alternative splicing of MAG and other myelin protein transcripts in the qk mouse is demonstrated. PMID- 11917127 TI - Elucidating the biosynthesis of 2-carboxyarabinitol 1-phosphate through reduced expression of chloroplastic fructose 1,6-bisphosphate phosphatase and radiotracer studies with 14CO2. AB - 2-carboxyarabinitol 1-phosphate limits photosynthetic CO2 assimilation at low light because it is a potent, naturally occurring inhibitor of ribulose 1,5 bisphosphate carboxylase/oxygenase. Evidence is presented that this inhibitor is derived from chloroplastic fructose 1,6-bisphosphate. First, transgenic plants containing decreased amounts of chloroplastic fructose 1,6-bisphosphate phosphatase contained increased amounts of fructose 1,6-bisphosphate and 2 carboxyarabinitol 1-phosphate and greatly increased amounts of the putative intermediates hamamelose and 2-carboxyarabinitol, which in some cases were as abundant as sucrose. Second, French bean leaves in the light were shown to incorporate 14C from 14CO2 sequentially into fructose 1,6-bisphosphate, hamamelose bisphosphate, hamamelose monophosphate, hamamelose, and 2 carboxyarabinitol. As shown previously, 14C assimilated by photosynthesis was also incorporated into 2-carboxyarabinitol 1-phosphate during subsequent darkness. PMID- 11917128 TI - Assembly of the neutrophil respiratory burst oxidase: a direct interaction between p67PHOX and cytochrome b558 II. AB - Activation of the phagocyte NADPH oxidase complex requires assembly of the cytosolic factors p47PHOX, p67PHOX, p40PHOX, and Rac with the membrane-bound cytochrome b558. We recently established a direct interaction between p67PHOX and cytochrome b558. In the present study, we show that removal of the C-terminal domain of p67PHOX increased its binding to cytochrome b558. Whereas phosphorylated p40PHOX alone did not bind to cytochrome b558, phosphorylated p47PHOX did, and, moreover, it allowed the binding of p40PHOX to the cytochrome. Furthermore, both increased the binding of p67PHOX) to the cytochrome. Phosphorylated p47PHOX thus appears to increase the binding of p67(PHOX) to cytochrome b558 by serving as an adapter, bringing p67PHOX into proximity with cytochrome b558, whereas phosphorylated p40(PHOX) may increase the binding by inducing a conformational change that allows p67PHOX to interact fully with cytochrome b558. PMID- 11917129 TI - Inhibition of APC-mediated proteolysis by the meiosis-specific protein kinase Ime2. AB - Proteolysis triggered by the anaphase-promoting complex (APC) is needed for sister chromatid separation and the exit from mitosis. APC is a ubiquitin ligase whose activity is tightly controlled during the cell cycle. To identify factors involved in the regulation of APC-mediated proteolysis, a Saccharomyces cerevisiae GAL-cDNA library was screened for genes whose overexpression prevented degradation of an APC target protein, the mitotic cyclin Clb2. Genes encoding G1, S, and mitotic cyclins were identified, consistent with previous data showing that the cyclin-dependent kinase Cdk1 associated with different cyclins is a key factor for inhibiting APC(Cdh1) activity from late-G1 phase until mitosis. In addition, the meiosis-specific protein kinase Ime2 was identified as a negative regulator of APC-mediated proteolysis. Ectopic expression of IME2 in G1 arrested cells inhibited the degradation of mitotic cyclins and of other APC substrates. IME2 expression resulted in the phosphorylation of Cdh1 in G1 cells, indicating that Ime2 and Cdk1 regulate APC(Cdh1) in a similar manner. The expression of IME2 in cycling cells inhibited bud formation and caused cells to arrest in mitosis. We show further that Ime2 itself is an unstable protein whose proteolysis occurs independently of the APC and SCF (Skp1/Cdc53/F-box) ubiquitin ligases. Our findings suggest that Ime2 represents an unstable, meiosis-specific regulator of APC(Cdh1). PMID- 11917130 TI - Chair-boat transitions in single polysaccharide molecules observed with force ramp AFM. AB - Under a stretching force, the sugar ring of polysaccharide molecules switches from the chair to the boat-like or inverted chair conformation. This conformational change can be observed by stretching single polysaccharide molecules with an atomic force microscope. In those early experiments, the molecules were stretched at a constant rate while the resulting force changed over wide ranges. However, because the rings undergo force-dependent transitions, an experimental arrangement where the force is the free variable introduces an undesirable level of complexity in the results. Here we demonstrate the use of force-ramp atomic force microscopy to capture the conformational changes in single polysaccharide molecules. Force-ramp atomic force microscopy readily captures the ring transitions under conditions where the entropic elasticity of the molecule is separated from its conformational transitions, enabling a quantitative analysis of the data with a simple two-state model. This analysis directly provides the physico-chemical characteristics of the ring transitions such as the width of the energy barrier, the relative energy of the conformers, and their enthalpic elasticity. Our experiments enhance the ability of single molecule force spectroscopy to make high-resolution measurements of the conformations of single polysaccharide molecules under a stretching force, making an important addition to polysaccharide spectroscopy. PMID- 11917131 TI - MYC overexpression imposes a nonimmunogenic phenotype on Epstein-Barr virus infected B cells. AB - Lymphoblastoid cell lines, generated by immortalization of normal B cells by Epstein-Barr virus (EBV) in vitro, have strong antigen-presenting capacity, are sensitive to EBV-specific cytotoxic T cells, and are highly allostimulatory in mixed lymphocyte culture. By contrast, EBV-positive Burkitt lymphoma (BL) cells are poor antigen presenters, are not recognized by EBV-specific cytotoxic T cells, and are poorly allostimulatory, which raises the question of whether immunological pressure exerted during BL pathogenesis in vivo has selected for a 'nonimmunogenic' tumor phenotype. The present work addresses this question by examining the immunogenicity/antigenicity of cell lines, generated by conversion of a conditionally immortalized lymphoblastoid cell line to permanent growth independent of EBV-latent proteins by introduction of a constitutively active or tetracycline-regulated c-myc gene (A1 and P493-6 cells, respectively). Compared with its parental lymphoblastoid cell line, A1 cells showed many of the features of the nonimmunogenic BL phenotype, namely poor allostimulatory activity, poor antigen-presenting function associated with impaired proteasomal activity, down regulation of peptide transporter, reduced HLA class I expression, and an inability to present endogenously expressed EBV-latent proteins to cytotoxic T cells. P493-6 cells, when grown in the presence of estrogen with the exogenous c myc gene switched off, were strongly immunogenic. The cells had lost their immunogenic potential, however, when grown on a c-myc-driven proliferation program in the absence of estrogen. Deregulation of c-myc, a step central to the development of uncontrolled BL cell growth in vivo, can thus impose a nonimmunogenic phenotype on proliferating human B cells in the absence of any immune pressure. PMID- 11917133 TI - Magnetic nanodots from atomic Fe: can it be done? AB - Laser focusing of Fe atoms offers the possibility of creating separate magnetic structures on a scale of 10 nm with exact periodicity. This can be done by using the parabolic minima of the potential generated by a standing light wave as focusing lenses. To achieve the desired 10-nm resolution, we need to suppress chromatic and spherical aberrations, as well as prevent structure broadening caused by the divergence of the incoming beam. Chromatic aberrations are suppressed by the development of a supersonic Fe beam source with speed ratio S = 11 +/- 1. This beam has an intensity of 3 x 10(15) atoms sr(-1) s(-1). The spherical aberrations of the standing light wave will be suppressed by aperturing with beam masks containing 100-nm slits at 744-nm intervals. The beam divergence can be reduced by application of laser cooling to reduce the transverse velocity. We have constructed a laser system capable of delivering over 500 mW of laser light at 372 nm, the wavelength of the (5)D(4) --> (5)F(5) atomic transition of (56)Fe we intend to use for laser cooling. Application of polarization spectroscopy to a hollow cathode discharge results in a locking system holding the laser continuously within 2 MHz of the desired frequency. PMID- 11917132 TI - DnaK dependence of mutant ethanol oxidoreductases evolved for aerobic function and protective role of the chaperone against protein oxidative damage in Escherichia coli. AB - The adhE gene of Escherichia coli encodes a multifunctional ethanol oxidoreductase (AdhE) that catalyzes successive reductions of acetyl-CoA to acetaldehyde and then to ethanol reversibly at the expense of NADH. Mutant JE52, serially selected for acquired and improved ability to grow aerobically on ethanol, synthesized an AdhE(A267T/E568K) with two amino acid substitutions that sequentially conferred improved catalytic properties and stability. Here we show that the aerobic growth ability on ethanol depends also on protection of the mutant AdhE against metal-catalyzed oxidation by the chaperone DnaK (a member of the Hsp70 family). No DnaK protection of the enzyme is evident during anaerobic growth on glucose. Synthesis of DnaK also protected E. coli from H2O2 killing under conditions when functional AdhE is not required. Our results therefore suggest that, in addition to the known role of protecting cells against heat stress, DnaK also protects numerous kinds of proteins from oxidative damage. PMID- 11917134 TI - A role for survivin in chemoresistance of endothelial cells mediated by VEGF. AB - Although standard anticancer chemotherapeutic drugs have been designed to inhibit the survival or growth of rapidly dividing tumor cells, it is possible to enhance the efficacy of such drugs by targeting the proliferating host endothelial cells (ECs) of the tumor vasculature. A theoretical advantage of this strategy lies in the possibility of circumventing, or significantly delaying, acquired drug resistance driven by the genetic instability of tumor cells. Here, we show that both vascular endothelial growth factor (VEGF) and basic fibroblast growth factor significantly reduce the pro-apoptotic potency of chemotherapy on both micro- and macrovascular ECs. This cytoprotection to drug toxicity was found to be phosphatidylinositol 3-kinase-dependent and could be recapitulated in the absence of VEGF by overexpressing the dominant-active form of the serine/threonine kinase protein kinase B/Akt. Downstream of phosphatidylinositol 3-kinase, we also show that survivin plays a pivotal role in VEGF-mediated EC protection by preserving the microtubule network. In this respect, its induction effectively protects ECs against chemotherapeutic damage, whereas overexpression of its dominant interfering mutant (C84A) abrogates the protective effects of VEGF. Accordingly, the potency of VEGF as a chemoprotectant was more pronounced with drugs that interfere with microtubule dynamics than those that damage DNA. These studies implicate a role for survivin up-regulation as a novel mechanism of EC drug "resistance" and support the notion that angiogenic factors that induce the expression of survivin may act to shield tumor ECs from the apoptotic effects of chemotherapy. Thus, exploiting chemotherapeutic drugs as antiangiogenics is likely to be compromised by the high concentrations of proangiogenic survival/growth factors present in the tumor microenvironment; targeting EC survival pathways should improve the antiangiogenic efficacy of antineoplastic agents, particularly microtubule-inhibitor drugs. PMID- 11917135 TI - Transcriptional activation of NtcA-dependent promoters of Synechococcus sp. PCC 7942 by 2-oxoglutarate in vitro. AB - The transcription factor NtcA is a global regulator of nitrogen homeostasis in cyanobacteria. It thus positively regulates the expression of genes related to nitrogen assimilation such as glnA (which encodes glutamine synthetase) and ntcA itself in response to nitrogen shortage or depletion. The binding of NtcA to the glnA and ntcA promoters of Synechococcus sp. PCC 7942 in vitro now has been shown to be enhanced by 2-oxoglutarate. In vitro analysis of gene transcription also revealed that the interaction of NtcA with its promoter element was not sufficient for activation of transcription, and 2-oxoglutarate was required for transcriptional initiation by NtcA. Given that the intracellular concentration of 2-oxoglutarate is inversely related to nitrogen availability, it is proposed that this metabolite functions as a signaling molecule that transmits information on cellular nitrogen status to NtcA and thereby regulates the transcription of genes related to nitrogen assimilation in cyanobacteria. PMID- 11917136 TI - Keratin binding to 14-3-3 proteins modulates keratin filaments and hepatocyte mitotic progression. AB - Keratin polypeptides 8 and 18 (K8/18) are the major intermediate filament proteins of simple-type epithelia. K18 Ser-33 phosphorylation regulates its binding to 14-3-3 proteins during mitosis. We studied the significance of keratin binding to 14-3-3 in transgenic mice that overexpress wild-type or Ser-33-->Ala (S33A) K18. In S33A but not wild-type K18-overexpressing mice, pancreatic acinar cell keratin filaments retracted from the basal nuclear region and became apically concentrated. In contrast, K18 S33A had a minimal effect on hepatocyte keratin filament organization. Partial hepatectomy of K18-S33A-overexpressing mice did not affect liver regeneration but caused limited mitotic arrest, accumulation of abnormal mitotic figures, dramatic fragmentation of hepatocyte keratin filaments, with retention of a speckled 14-3-3zeta mitotic cell nuclear staining pattern that usually becomes diffuse during mitosis. Hence, K18 Ser-33 phosphorylation regulates keratin filament organization in simple-type epithelia in vivo. Keratin binding to 14-3-3 may partially modulate hepatocyte mitotic progression, in association with nuclear redistribution of 14-3-3 proteins during mitosis. PMID- 11917137 TI - The homeobox gene BREVIPEDICELLUS is a key regulator of inflorescence architecture in Arabidopsis. AB - Flowering plants display a remarkable range of inflorescence architecture, and pedicel characteristics are one of the key contributors to this diversity. However, very little is known about the genes or the pathways that regulate pedicel development. The brevipedicellus (bp) mutant of Arabidopsis thaliana displays a unique phenotype with defects in pedicel development causing downward pointing flowers and a compact inflorescence architecture. Cloning and molecular analysis of two independent mutant alleles revealed that BP encodes the homeodomain protein KNAT1, a member of the KNOX family. bp-1 is a null allele with deletion of the entire locus, whereas bp-2 has a point mutation that is predicted to result in a truncated protein. In both bp alleles, the pedicels and internodes were compact because of fewer cell divisions; in addition, defects in epidermal and cortical cell differentiation and elongation were found in the affected regions. The downward-pointing pedicels were produced by an asymmetric effect of the bp mutation on the abaxial vs. adaxial sides. Cell differentiation, elongation, and growth were affected more severely on the abaxial than adaxial side, causing the change in the pedicel growth angle. In addition, bp plants displayed defects in cell differentiation and radial growth of the style. Our results show that BP plays a key regulatory role in defining important aspects of the growth and cell differentiation of the inflorescence stem, pedicel, and style in Arabidopsis. PMID- 11917138 TI - Complementary advantageous substitutions in the evolution of an antiviral RNase of higher primates. AB - An improved understanding of the evolution of gene function at the molecular level may provide significant insights into the origin of biological novelty and adaptation. With the approach of ancestral protein reconstruction, we here address the question of how a dramatically enhanced ribonucleolytic activity and the related antiviral activity evolved in a recently duplicated ribonuclease (eosinophil-derived neurotoxin) gene of higher primates. We show that the mother gene of the duplicated genes had already possessed a weak antiviral activity before duplication. After duplication, substitutions at two interacting sites (Arg-64-->Ser and Thr-132-->Arg) resulted in a 13-fold enhancement of the ribonucleolytic activity of eosinophil-derived neurotoxin. These substitutions are also necessary for the potent antiviral activity, with contributions from additional amino acid changes at interacting sites. Our observation that a change in eosinophil-derived neurotoxin function occurs only when both interacting sites are altered indicates the importance of complementary substitutions in protein evolution. Thus, neutral substitutions are not simply "noises" in protein evolution, as many have thought. They may play constructive roles by setting the intramolecular microenvironment for further complementary advantageous substitutions, which can lead to improved or altered function. Overall, our study illustrates the power of the "paleomolecular biochemistry" approach in delineating the complex interplays of amino acid substitutions in evolution and in identifying the molecular basis of biological innovation. PMID- 11917139 TI - Helicogenicity of solvents in the conformational equilibrium of oligo(m-phenylene ethynylene)s: implications for foldamer research. AB - A (R)-binaphthol tethered bis-hexameric oligo(m-phenylene ethynylene) foldamer was examined in 30 solvents to correlate the unfolded-folded conformational equilibrium to bulk solvent properties and specific solvent-chain interactions. The oligomer is soluble in a variety of solvents of intermediate polarity, with the majority of these solvents being helicogenic. The amphiphilic nature of the chain allows the solvophobic backbone to be solubilized in a wide range of solvents through the polar triethylene glycol side chains. As demonstrated through UV and CD spectroscopic experiments, the helical conformation is increasingly stabilized with increasing solvent polarity in the absence of specific solvent-chain interactions. Surprisingly, very few solvents are capable of fully denaturing the helix, indicating the strength of the solvophobic driving forces in this cooperative system. The folding reaction for this amphiphilic oligomer can be described as a compromise in solubility properties, where chains collapse intramolecularly into helical conformations to minimize solvent-backbone contacts while maintaining favorable solvent-side chain interactions for solvation. In terms of mimicking the properties of biomacromolecules, foldamers using solvophobic driving forces must be tempered with functionalities that promote solubility of the folded state while at the same time allowing access to the unfolded state through the use of denaturants. PMID- 11917140 TI - Cell membrane GM1 ganglioside is a functional coreceptor for fibroblast growth factor 2. AB - Free gangliosides bind fibroblast growth factor 2 (FGF2), thus preventing cell interaction and biological activity of the growth factor in endothelial cells. Here we investigated the role of cell-associated gangliosides in mediating the biological activity of FGF2. Treatment of endothelial cells of different origin with the ganglioside biosynthesis inhibitors fumonisin B1, D-threo-1-phenyl-2 decanoylamino-3-morpholino-1-propanol or D-1-threo-1-phenyl-2-hexa-decanoylamino 3-pyrrolidino-1-propanol-HCl, impairs their capacity to proliferate when exposed to FGF2. Also, the mitogenic activity of FGF2 is inhibited by the GM1-binding cholera toxin B subunit (CTB). Conversely, overloading of endothelial GM 7373 cell membranes with exogenous GM1 causes a 10-fold increase of the mitogenic potency of FGF2. 125I-FGF2 binds to cell membrane GM1 (K(d) = 3 nM) in complex ganglioside/heparan sulfate-deficient Chinese hamster ovary (CHO)-K1-pgsA745 cell mutants that were overloaded with exogenous GM1. Moreover, FGF2 competes with FITC-CTB for the binding to cell membrane GM1 in different CHO cell lines independently of their capacity to express heparan sulfate proteoglycans. Conversely, CTB inhibits cell proliferation triggered by FGF2 in CHO cells overexpressing the tyrosine kinase FGF receptor 1. Finally, GM1-overloading confers to FGF receptor 1-transfected, complex ganglioside-deficient CHO-K1 cell mutants the capacity to proliferate when stimulated by FGF2. This proliferation is inhibited by CTB. Cell proliferation triggered by serum or by phorbol 12 myristate 13-acetate is instead independent of the cell membrane ganglioside milieu. In conclusion, cell membrane GM1 binds FGF2 and is required for the mitogenic activity of the growth factor. Our data indicate that cell-associated gangliosides may act as functional FGF2 co-receptors in different cell types. PMID- 11917143 TI - Use of covariance analysis for the prediction of structural domain boundaries from multiple protein sequence alignments. AB - Current methods for identification of domains within protein sequences require either structural information or the identification of homologous domain sequences in different sequence contexts. Knowledge of structural domain boundaries is important for fold recognition experiments and structural determination by X-ray crystallography or nuclear magnetic resonance spectroscopy using the divide-and-conquer approach. Here, a new and conceptually simple method for the identification of structural domain boundaries in multiple protein sequence alignments is presented. Analysis of covariance at positions within the alignment is first used to predict 3D contacts. By the nature of the domain as an independent folding unit, inter-domain predicted contacts are fewer than intra domain predicted contacts. By analysing all possible domain boundaries and constructing a smoothed profile of predicted contact density (PCD), true structural domain boundaries are predicted as local profile minima associated with low PCD. A training data set is constructed from 52 non-homologous two domain protein sequences of known 3D structure and used to determine optimal parameters for the profile analysis. The alignments in the training data set contained 48 +/- 17 (mean +/- SD) sequences and lengths of 257 +/- 121 residues. Of the 47 alignments yielding predictions, 35% of true domain boundaries are predicted to within 15 amino acids by the local profile minimum with the lowest profile value. Including predictions from the second- and third-lowest local minima increases the correct domain boundary coverage to 60%, whereas the lowest five local minima cover 79% of correct domain boundaries. Through further profile analysis, criteria are presented which reliably identify subsets of more accurate predictions. Retrospective analysis of CASP3 targets shows predictions of sufficient accuracy to enable dramatically improved fold recognition results. Finally, a prediction is made for geminivirus AL1 protein which is in full agreement with biochemical data, yielding a plausible, novel threading result. PMID- 11917141 TI - The accessory subunit of DNA polymerase gamma is essential for mitochondrial DNA maintenance and development in Drosophila melanogaster. AB - DNA polymerase gamma, Pol gamma, is the key replicative enzyme in animal mitochondria. The Drosophila enzyme is a heterodimer comprising catalytic and accessory subunits of 125 kDa and 35 kDa, respectively. Both subunits have been cloned and characterized in a variety of model systems, and genetic mutants of the catalytic subunit were first identified in Drosophila, as chemically induced mutations that disrupt larval behavior (tamas). Mutations in the gene encoding the accessory subunit have not yet been described in any organism. Here, we report the consequences of null mutations upon mitochondrial DNA (mtDNA) replication and morphology, cell proliferation, and organismal viability. Mutations in the accessory subunit cause lethality during early pupation, concomitant with loss of mtDNA and mitochondrial mass, and reduced cell proliferation in the central nervous system. Surprisingly, the function of the central nervous system and muscle, as assessed in a locomotion assay, are only marginally affected. This finding is in contrast to our previous findings that disruption in the function of the catalytic subunit causes severe reduction in larval locomotion. We discuss our results in the context of current hypotheses for the function of the accessory subunit in mtDNA replication. PMID- 11917144 TI - A Fourier analysis of symmetry in protein structure. AB - The score matrix from a structure comparison program (SAP) was used to search for repeated structures using a Fourier analysis. When tested with artificial data, a simple Fourier transform of the smoothed matrix provided a clear signal of the repeat periodicity that could be used to extract the repeating units with the SAP program. The strength of the Fourier signal was calibrated against the signal from model proteins. The most useful of these was the novel random-walk approach employed to generate realistic 'fake' structures. On the basis of these it was possible to conclude that only a small proportion of protein structures have an unexpected degree of symmetry. Artificially generated 'ideal' folds provided an upper limit on the strength of signal that could be expected from a 'perfectly' repeating compact structure. Unexpectedly, some of the very regular beta propellor folds attained the same strength but the majority of symmetric structures lay below this region. When native proteins were ranked by the power of their spectrum a wide variety of fold types were seen to score highly. In the betaalpha class, these included the globular betaalpha proteins and the more repetitive leucine-rich betaalpha folds. In the all-beta class; beta-propellors, beta-prisms and beta-helices were found as well as the more globular gamma crystalin domains. When this ranked list was filtered to remove proteins that contained detectable internal sequence similarity (using the program REPRO), the list became exclusively composed of just globular betaalpha class proteins and in the top 50 re-ranked proteins, only a single 4-fold propellor structure remained. PMID- 11917142 TI - Genetic evidence for Shc requirement in TCR-induced c-Rel nuclear translocation and IL-2 expression. AB - Shc, a prototypic adapter molecule, has been implicated in T cell receptor (TCR) signal transduction, but its role has not been identified clearly. Here we report that Shc is essential for TCR-induced IL-2 production but is dispensable for CD69 or CD25 expression. Engagement of TCR in mutant Jurkat T cells lacking Shc fails to produce IL-2 because of impaired mitogen-activated protein kinase activation. Activation of c-Rel, a transcription factor essential for IL-2 expression, was impaired also. In contrast, activation of nuclear factor of activated T cell and expression of CD69/CD25 were comparable between the mutant and wild-type Jurkat cells. These defects were rescued by expression of exogenous Shc. Activation of c Rel using the estrogen receptor fusion protein restored the activation of the IL 2 promoter in an estrogen-dependent manner. These results show that Shc plays an essential role in the TCR-induced activation of c-Rel and the IL-2 promoter. PMID- 11917146 TI - In silico identification, structure prediction and phylogenetic analysis of the 2'-O-ribose (cap 1) methyltransferase domain in the large structural protein of ssRNA negative-strand viruses. AB - The Escherichia coli RrmJ gene product has recently been shown to be the 23S rRNA:U2552 specific 2'-O-ribose methyltransferase (MTase) (RrmJ). Its structure has been solved and refined to 1.5 A resolution, demonstrating conservation of the three-dimensional fold and key catalytic side chains with the vaccinia virus VP39 protein, which functions as an mRNA 5'm(7)G-cap-N-specific 2'-O-ribose MTase. Using the amino acid sequence of RrmJ as an initial probe in an iterative search of sequence databases, we identified a homologous domain in the sequence of the L protein of non-segmented, negative-sense, single-stranded RNA viruses. The plausibility of the prediction was confirmed by homology modeling and checking whether important residues at substrate/ligand-binding sites were conserved. The predicted structural compatibility and the conservation of the active site between the novel putative MTase domain and genuine 2'-O-ribose MTases, together with the available results of biochemical studies, strongly suggest that this domain is a 5'm(7)G-cap-N-specific 2'-O-ribose MTase (i.e. the cap 1 MTase). Evolutionary relationships between these proteins are also discussed. PMID- 11917145 TI - Aromatic-aromatic interactions in and around alpha-helices. AB - To understand the role of aromatic-aromatic interactions in imparting specificity to the folding process, the geometries of four aromatic residues with different sequence spacing, located in alpha-helices or five residues from helical ends, interacting with each other have been elucidated. The geometry is found to depend on the sequence difference. Specific interactions (C-H...pi and N-H...pi) which result from this geometry may cause a given pair of residues (such as Phe-His) with a particular sequence difference to occur more than expected. The most conspicuous residue in an aromatic pair in the context of helix stability is His, which is found at the last (C1) position or the two positions (Ncap and Ccap) immediately flanking the helix. An alpha-helix and a contiguous 3(10)-helix or two helices separated by a non-helical residue can have interacting aromatic pairs, the geometry of interaction and the relative orientation between the helices being rather fixed. Short helices can also have interacting residues from either side. PMID- 11917147 TI - A molecular dynamics study of an L-type calcium channel model. AB - In this work, we propose a molecular model of the L-type calcium channel pore from the human cardiac alpha1 subunit. Four glutamic acid residues, the EEEE locus, located at highly conserved P loops (also called SS1-SS2 segments) of the alpha1 subunit, molecularly express the calcium channel selectivity. The proposed alpha-helix structure for the SS1 segment, analyzed through molecular dynamics simulations in aqueous-phase, was validated by the plotting of Ramachandran diagrams for the averaged structures and by the analysis of i and i + 4 helical hydrogen bonding between the amino acid residues. The results of the simulation of the calcium channel model with one and two Ca2+ ions at the binding site are in accordance with mutation studies which suggest that the EEEE locus in the L type calcium channel must form a single high-affinity binding site. These results suggest that the Ca2+ permeation through the channel would be derived from competition between two ions for the only high-affinity binding site. Furthermore, the experimentally observed blocking of the Na+ flux at micromolar Ca2+ concentrations, probably due to the occupancy of the single high-affinity binding site for one Ca2+, was also reproduced by our model. PMID- 11917148 TI - Development of selectivity of alpha1-antitrypsin variant by mutagenesis in its reactive site loop against proprotein convertase. A crucial role of the P4 arginine in PACE4 inhibition. AB - PACE4, furin and PC6 are Ca2+-dependent serine endoproteases that belong to the subtilisin-like proprotein convertase (SPC) family. Recent reports have supported the involvement of these enzymes in processing of growth/differentiation factors, viral replication, activation of bacterial toxins and tumorigenesis, indicating that these enzymes are a fascinating target for therapeutic agents. In this work, we evaluated the sensitivity and selectivity of three rat alpha1-antitrypsin variants which contained RVPR352, AVRR352 and RVRR352, respectively, within their reactive site loop using both inhibition of enzyme activity toward a fluorogenic substrate in vitro and formation of a SDS-stable protease/inhibitor complex ex vivo. The RVPR variant showed relatively broad selectivity, whereas the AVRR and RVRR variants were more selective than the RVPR variant. The AVRR variant inhibited furin and PC6 but not PACE4. This selectivity was further confirmed by complex formation and inhibition of pro-complement C3 processing. On the other hand, although the RVRR variant inhibited both PACE4 and furin effectively, it needed a 600-fold higher concentration than the RVPR variant to inhibit PC6 in vitro. These inhibitors will be useful tools in helping us to understand the roles of PACE4, furin and PC6. PMID- 11917149 TI - Alteration of the specificity of the cofactor-binding pocket of Corynebacterium 2,5-diketo-D-gluconic acid reductase A. AB - The NADPH-dependent 2,5-diketo-D-gluconic acid (2,5-DKG) reductase enzyme is a required component in some novel biosynthetic vitamin C production processes. This enzyme catalyzes the conversion of 2,5-DKG to 2-keto-L-gulonic acid, which is an immediate precursor to L-ascorbic acid. Forty unique site-directed mutations were made at five residues in the cofactor-binding pocket of 2,5-DKG reductase A in an attempt to improve its ability to use NADH as a cofactor. NADH is more stable, less expensive and more prevalent in the cell than is NADPH. To the best of our knowledge, this is the first focused attempt to alter the cofactor specificity of a member of the aldo-keto reductase superfamily by engineering improved activity with NADH into the enzyme. Activity of the mutants with NADH or NADPH was assayed using activity-stained native polyacrylamide gels. Eight of the mutants at three different sites were identified as having improved activity with NADH. These mutants were purified and subjected to a kinetic characterization with NADH as a cofactor. The best mutant obtained, R238H, produced an almost 7-fold improvement in catalysis with NADH compared with the wild-type enzyme. Surprisingly, most of this catalytic improvement appeared to be due to an improvement in the apparent kcat for the reaction rather than a large improvement in the affinity of the enzyme for NADH. PMID- 11917150 TI - Engineering of multiple arginines into the Ser/Thr surface of Trichoderma reesei endo-1,4-beta-xylanase II increases the thermotolerance and shifts the pH optimum towards alkaline pH. AB - We studied the effects of increase in the number of surface arginines on the enzyme activity and stability of Trichoderma reesei endo-1,4-beta-xylanase II. The number of arginines was increased in two mutant series. The first set contained six arginines on different sides of the protein surface. These arginines had no significant effect on the thermostability. However, the optimal pH region became narrower. Another series of five arginines was engineered into the 'Ser/Thr surface', formed of part of the double-layered beta-sheet located on one side of the 'right-hand-like' xylanase. These mutations shifted the activity profile to the alkaline region by approximately 0.5-1.0 pH units. In addition, the arginines on the Ser/Thr surface increased the enzyme activity at high temperature, although the enzyme stability in the absence of substrate decreased significantly at 50-55 degrees C. In the presence of the substrate, the thermostability increased 4-5-fold at 60-65 degrees C. Thus, the substrate neutralized the destabilizing effect of Ser/Thr surface arginines and revealed a stabilizing effect of the same mutations. The stabilizing effect of arginines at high temperatures was seen clearly only when five arginines were introduced into the Ser/Thr surface. PMID- 11917151 TI - Modifying the chain-length selectivity of the lipase from Burkholderia cepacia KWI-56 through in vitro combinatorial mutagenesis in the substrate-binding site. AB - The mature lipase of Burkholderia cepacia KWI-56 was synthesized in an enzymatically active form using an in vitro Escherichia coli S30 coupled transcription/translation system by expressing the mature lipase gene (rlip) in the presence of its specific activator. To investigate the substrate specificity of the lipase comprehensively, a large number of mutant lipases were constructed and analyzed in a high throughput manner by combining overlapping PCR and in vitro protein synthesis. In this paper, Phe119 and Leu167, which are located in the acyl portion of the substrate-binding pocket of the lipase of B.cepacia KWI 56, were substituted with six hydrophobic amino acid residues by the in vitro combinatorial mutagenesis. The wild-type and 35 mutant genes amplified by PCR were directly used as templates for the in vitro transcription/translation. The acyl chain-length selectivity of the in vitro expressed lipases against p nitrophenyl butyrate, p-nitrophenyl caprylate and p-nitrophenyl palmitate, was compared by their relative hydrolysis rates. Two mutant lipases, L167V and F119A/L167M, which showed a significant shift in substrate selectivity were further expressed in vivo and refolded in vitro. It was found that L167V raised its preference for the short-chain ester, whereas F119A/L167M improved its selectivity for the long-chain ester. PMID- 11917152 TI - Gene expression response to misfolded protein as a screen for soluble recombinant protein. AB - Proper protein folding is key to producing recombinant proteins for structure determination. We have examined the effect of misfolded recombinant protein on gene expression in Escherichia coli. Comparison of expression patterns indicates a unique set of genes responding to translational misfolding. The response is in part analogous to heat shock and suggests a translational component to the regulation. We have further utilized the expression information to generate reporters responsive to protein misfolding. These reporters were used to identify properly folded recombinant proteins and to create soluble domains of insoluble proteins for structural studies. PMID- 11917154 TI - Visualizing cellular phosphoinositide pools with GFP-fused protein-modules. AB - Inositol phospholipids are well known for their pivotal role in calcium signaling as precursors of important second messengers generated in response to various stimuli. However, over the last 10 years, inositides have also emerged as universal signaling components present in virtually every membrane of eukaryotic cells. These lipids are locally produced and degraded by the numerous inositide kinase and phosphatase enzymes, to control the recruitment and activity of protein signaling complexes in specific membrane compartments. The spatial and temporal constraints imposed on changes in cellular inositides pose new challenges in finding experimental techniques through which such changes can be examined. Taking advantage of the protein domains selected by evolution to recognize cellular phosphoinositides, we have created fluorescent molecules by fusing these domains to the improved version of green fluorescent protein (EGFP); the distribution of these fusion proteins can be followed within live cells, thereby reporting on changes in phosphoinositides. Although this technique is one of the few that provide information on phosphoinositide dynamics in live cells with subcellular resolution and has rapidly gained popularity, it also has limitations that need to be taken into account when interpreting the data. Here, we summarize our experience in designing and using these constructs and review our position concerning the interpretation of the data obtained by this technique. PMID- 11917155 TI - Confocal imaging of subcellular Ca2+ concentrations using a dual-excitation ratiometric indicator based on green fluorescent protein. AB - Dual-excitation ratiometric dyes are excited alternately at two different wavelengths, but the emission is collected at a single fixed wavelength. Therefore, the pair of intensity measurements must be collected sequentially. Ratiometric-pericam is a fluorescent Ca(2+) indicator based on a chimeric fusion protein of circularly permuted green fluorescent protein and calmodulin. Upon binding to calcium, its excitation peak shifts from 415 nm to 494 nm. Ca(2+) imaging using ratiometric-pericam was thought to be inadequate to follow very fast Ca(2+) dynamics or Ca(2+) changes in highly motile cell samples; however, we describe a technique that allows high spatial and time resolution of images acquired with ratiometric-pericam. To obtain confocal images of Ca(2+) using ratiometric-pericam, we established a system in which two laser beams (excitation 408 nm and 488 nm) are alternated on every scanning line under the control of two acousto-optic tunable filters. This system increases the rate at which ratio measurements are done to 200 Hz, and provides confocal images at 1 to 10 Hz depending on the image size. The ratio images are free from noise caused by the fluctuation of laser power, because the system is equipped with a violet laser diode (408 nm) and a diode-pumped solid-state laser (488 nm), both of which are stable. We visualized the dynamic propagation of Ca(2+) waves from the cytosol to the nucleus and changes in Ca(2+) concentrations in motile mitochondria of HeLa cells. We demonstrate that this new confocal imaging system expands the range of potential applications of ratiometric-pericam and other dual-excitation ratiometric indicators. PMID- 11917156 TI - Mouse models of human disease: lessons learned and promises to come. PMID- 11917157 TI - Mouse models for disorders of mitochondrial fatty acid beta-oxidation. AB - Mitochondrial beta-oxidation of fatty acids is vital for energy production in periods of fasting and other metabolic stress. Human patients have been identified with inherited disorders of mitochondrial beta-oxidation of fatty acids with enzyme deficiencies identified at many of the steps in this pathway. Although these patients exhibit a range of disease processes, Reye-like illness (hypoketotic-hypoglycemia, hyperammonemia and fatty liver) and cardiomyopathy are common findings. There have been several mouse models developed to aid in the study of these disease conditions. The characterized mouse models include inherited deficiencies of very long-chain acyl-CoA dehydrogenase, long-chain acyl CoA dehydrogenase, short-chain acyl-CoA dehydrogenase, mitochondrial trifunctional protein-alpha, and medium-/short-chain hydroxyacyl-CoA dehydrogenase. Mouse mutants developed, but presently incompletely characterized as models, include carnitine palmitoyltransferase-1a and medium-chain acyl-CoA dehydrogenase deficiencies. In general, the mouse models of disorders of mitochondrial fatty acid beta-oxidation have shown clinical signs that include Reye-like syndrome and cardiomyopathy, and many are cold intolerant. It is expected that these mouse models will provide vital contributions in understanding the mechanisms of disease pathogenesis of fatty acid oxidation disorders and the development of appropriate treatments and supportive care. PMID- 11917158 TI - The mousetrap: what we can learn when the mouse model does not mimic the human disease. AB - In recent years, mouse models for human metabolic diseases have become commonplace because the information gained from in vivo study of biochemical pathways is invaluable, and many metabolic diseases are relatively easy to recreate in mice through gene knockout technology in embryonic stem cells. In certain cases, however, the knockout mice may reproduce only some of the human disease phenotype, may be more severely affected than human cases, or may have no clinical phenotype at all. Under these circumstances, the disease pathology can become more complex, causing the researcher to evaluate basic differences in mouse and human biology as well as questions of genetic background, alternate pathways, and possible gene interactions. This review is a brief analysis of gene knockout models for Lesch-Nyhan syndrome, Lowe syndrome, X-linked adrenoleukodystrophy, Fabry disease, galactosemia, glycogen storage disease type II, metachromatic leukodystrophy, and Tay-Sachs disease, which produce a biochemical model of disease but often do not reproduce clinical symptoms. These mice may be useful for studying the biochemical and physiological pathways in which certain metabolites function toward embryonic and fetal development, as well as specific functions in various organs, and they may provide an inexpensive and useful model system for development of new therapeutic techniques. PMID- 11917159 TI - Complexities of cancer research: mouse genetic models. AB - Cancer susceptibility is a complex interaction of an individual's genetic composition and environmental exposures. Huge strides have been made in understanding cancer over the past 100 yr, from recognition of cancer as a genetic disease, to identification of specific carcinogens, isolation of oncogenes, and recognition of tumor suppressors. A tremendous amount of knowledge has accumulated about the etiology of cancer. Cancer genetics has played a significant role in these discoveries. Analysis of high-risk familial cancers has led to the discovery of new tumor suppressor genes and important cancer pathways. These families, however, represent only a small fraction of cancer in the general population. Most cancer is instead probably the result of an intricate interaction of polymorphic susceptibility genes with the sea of environmental exposures that humans experience. Although the central cadre of cancer genes is known, little is understood about the peripheral genes that likely comprise the polymorphic susceptibility loci. The challenge for cancer genetics is therefore to move forward from the mendelian genetics of the rare familial cancer syndromes into the field of quantitative trait loci, susceptibility factors, and modifier genes. By identifying the genes that modulate an individual's susceptibility to cancer after an environmental exposure, researchers will be able to gain important insights into human biology, cancer prevention, and cancer treatment. This article summarizes the current state of quantitative trait genetic analysis and the tools, both proven and theoretical, that may be used to unravel one of the great challenges in cancer genetics. PMID- 11917160 TI - Mouse models of Alzheimer's disease: a quest for plaques and tangles. AB - Many genetically altered mice have been designed to help understand the role of specific gene mutations in the pathogenesis of Alzheimer's disease (AD) based on the realization that specific mutations in the genes for amyloid precursor protein--the presenilins and tau--are associated with early-onset familial AD or, in the case of tau mutations, other neurodegenerative diseases with neurofibrillary tangles. However, attempts to reproduce the neuropathology of AD in the mouse have been frustrating. Transgenic designs emphasizing amyloid precursor protein produced mice that develop amyloid plaques, but neurodegeneration and neurofibrillary tangles failed to form. Strategies emphasizing tau resulted in increased phosphorylation of tau and tangle formation, although amyloid plaques were absent. Nevertheless, crossing transgenic animals expressing mutated tau and amyloid precursor protein has produced a mouse that closely recapitulates the neuropathology of AD. A review of the various murine models, their role in understanding the pathogenesis of AD and their use in testing therapeutic regimens, is provided. PMID- 11917161 TI - Welfare issues of genetically modified animals. AB - Genetically engineered animals have opened new frontiers in the study of physiology and disease processes. Mutant animals offer more accurate disease models and increased precision for pathogenesis and treatment studies. Their use offers hope for improved therapy to patients with conditions that currently have poor or ineffective treatments. These advantages have fostered an increase in studies using mice in recent years, a development viewed with alarm by those who oppose the use of animals in research. Scientists point out that the mice are replacing more sentient species, such as nonhuman primates, and are increasing the quality of research being conducted. They assert that study of genetically engineered animals will eventually permit decreases in numbers of animals used in research. Nevertheless, the increase in use of genetically altered animals presents many challenges in reviewing protocols and providing care. Identification and resolution of any welfare problems is a responsibility that is shared by institutional animal care and use committee, veterinary, animal care, and research staffs. To identify potential welfare concerns, a database such as TBASE () can be searched to learn what has been reported for established mutant lines. In addition, newly created lines should be monitored by a surveillance system and have phenotype assessment to identify the effects of altering the genome. Methods of ensuring welfare can include treatment of conditions produced, restriction of gene expression to tissues of interest or to certain time periods, and establishment of endpoints for removing animals from a study before problems appear. PMID- 11917162 TI - The future of animal research. PMID- 11917163 TI - Cutaneous T-cell lymphoproliferative disorders: approach for the surgical pathologist: recent advances and clarification of confused issues. AB - Cutaneous T-cell lymphoproliferative disorders (CTCLs) remain a subject of confusion and controversy. In this review, the authors discuss diagnostic criteria and classification, including the role of immunohistochemistry and gene rearrangement studies. In addition, cutaneous T-cell pseudolymphomas, the current status of parapsoriasis and other premalignant syndromes, and the clinicopathological variants of mycosis fungoides are discussed. CD30-positive lymphoproliferative disorders and a number of rare variants of CTCL including granulamatous slack skin, subcutaneous (panniculitic) T-cell lymphoma, gamma delta cutaneous lymphoma, NK/NK-like T-cell lymphoma, and primary cutaneous CD8 positive epidermotropic cytotoxic T-cell lymphoma are also considered. PMID- 11917164 TI - Differential diagnosis in nephropathology: an immunofluorescence-driven approach. AB - Evaluation of a renal biopsy can be intimidating, because observational data from histologic stains, direct immunofluorescence (IF), and electron microscopy must be integrated with clinical and laboratory data to arrive at the correct diagnosis. Numerous diagnostic categories usually merit consideration. The typical strategy to diagnose renal disease is heavily weighted toward analysis of the histologic pattern of glomerular injury. This focus on histology is understandable; histologic evaluation was the first analytic probe available and has driven the formulation of classification schemes. Evaluation of IF before review of the light microscopy facilitates diagnostic evaluation, because the possibilities are immediately reduced. This simplifies the crucial final step clinical and histologic correlation. In this review, I discuss the information content of IF and provide an algorithmic approach to the major renal diseases based on an IF-driven dissection of the differential diagnosis. PMID- 11917165 TI - Morphologic and molecular prognostic markers in prostate cancer. AB - In this review, a series of traditional morphology-based measurements, relatively well-documented ancillary biomarkers, and emerging molecular assays are evaluated for their relative ability to predict prognosis in prostate cancer. Prognostic factors that have achieved widespread use and are classified as category I by the College of American Pathologists' Solid Tumor Prognostic Factor Consensus Conference are compared with newer tests that are beginning to be used in clinical practice (category II) and emerging molecular-based assays that have yet to be widely validated in the published literature or in clinical trials (category III). PMID- 11917166 TI - Dismantling the germinal center: comparing the processes of transformation, regression, and fragmentation of the lymphoid follicle. AB - The pathologic and immunologic features of the formation of the germinal center have been extensively studied. The process of dissolution of the germinal center is not as well understood. Different patterns of germinal-center breakdown are commonly encountered in diagnostic lymph node biopsy specimens and frequently present difficulties in diagnosis. The current immunologic understanding of germinal-center dynamics is reviewed and correlated with the histologic and immunophenotypic features of three broad classes of germinal center dissolution, namely progressive transformation, regression, and follicle fragmentation. The author suggests that these different patterns represent alternate responses to antigenic stimulation. The relationships of progressive transformation to Hodgkin's disease and of follicular regression to Castleman's disease are discussed. PMID- 11917168 TI - Applying robotic assistance to laparoscopic cholecystectomy. PMID- 11917170 TI - Laser-assisted in situ keratomileusis complications in diabetes mellitus. AB - PURPOSE: To report the incidence of complications and the refractive results in patients with diabetes mellitus treated with laser-assisted in situ keratomileusis (LASIK). METHODS: A retrospective review of the charts, focusing on 6-month postoperative data, was performed on 30 eyes from patients with diabetes and 150 age- and gender-matched control eyes operated on during the same period. The incidence of complications and postoperative refractive results were compared. The data analyzed include UCVA, spherical equivalent, astigmatism power, astigmatism axis, and vector astigmatism change. RESULTS: Diabetic eyes treated with LASIK had an overall complication rate of 47% compared with the control population complication incidence of 6.9% (p< 0.01). The most frequent complications occurring in the diabetic population are punctate epithelial erosions and persistent epithelial defects. Spherical correction change was -4.64 diopters (D) for diabetic eyes and -4.98 D for control eyes (p= 0.49). Mean spherical equivalent change was -4.69 D for diabetic eyes and -4.75 D for control eyes (p= 0.9). Mean change in uncorrected visual acuity (LogMAR) was 1.5 for diabetic eyes and 1.65 for control eyes (p= 0.18). Mean astigmatism magnitude change was 0.31 in diabetic eyes and 0.57 in control eyes (p= 0.12). Mean vector corrected astigmatism change was 0.97 for diabetic eyes and 1.12 for control eyes (p= 0.31). Mean vector-corrected astigmatism axis for patients with diabetes was 18.17 for diabetic eyes and 6.20 for control eyes (p= 0.30). CONCLUSION: Patients with diabetes who undergo LASIK are at a significantly higher risk of developing postoperative epithelial complications. In addition, this study revealed poorer refractive results in the eyes of patients with diabetes treated with LASIK. PMID- 11917171 TI - Quantitative descriptors of corneal topography that influence soft toric contact lens fitting. AB - PURPOSE: Empirical soft toric contact lens fitting based on manifest refraction and keratometry often presents unanticipated fitting and power errors upon initial lens dispensing. However, corneal topography may provide features that influence soft toric lens performance, flexure, and back vertex power in situ, which may assist in improved fitting guidelines. In this study, quantitative topographic descriptors were generated and analyzed as potential variables in predicting soft toric fitting success. METHODS: One hundred five eyes of 54 patients were empirically fit with back surface toric, prism ballasted, soft contact lenses after videokeratography was performed with the EyeSys 2000 (v. 4.0) or Humphrey Atlas (v. A6) instrument. Custom software was written to generate 54 separate quantitative descriptors of shape and astigmatism from the raw data files. A logistic regression was used to determine which variables significantly contributed to a successful or failed fit. RESULTS: Two types of empirical fitting failures were identified: loose fit (n = 15) and power errors (n = 17). The following variables were associated with a fitting failure: flat simulated keratometry (SimKf2b) within the central 3 mm zone, steep simulated keratometry (SimKs2b) within the central 3 mm zone, a difference between central and peripheral flat meridian axis (DIFFAXIS), and a difference between central and peripheral astigmatism (DIFFASTIG). For fitting failures caused by power errors, a larger steep SimKs2b (p< 0.01) and smaller DIFFAXIS (p< 0.05) were associated with a failed fit. For failures caused by physical fit of a selected base curve, a smaller DIFFAXIS (p< 0.05), larger steep SimKs2b (p< 0.05), and larger DIFFASTIG (p< 0.01) were associated with a failed fit. CONCLUSIONS: Novel quantitative descriptors of corneal shape and toricity derived from topography are associated with empirical soft toric contact lens fitting failures. Future algorithms or recommendations for improved soft toric lens selection may be derived from such indices to develop a predictive model for successful soft toric lens fitting using corneal topography data. PMID- 11917172 TI - Fourier series harmonic analysis of corneal topography following suture removal after penetrating keratoplasty. AB - PURPOSE: To evaluate quantitatively the influence of suture removal after penetrating keratoplasty on corneal refraction including irregular astigmatism. METHODS: Subjects were 42 eyes of 39 patients with clear corneal grafts who underwent suture removal after penetrating keratoplasty. Videokeratography data taken before and after suture removal were decomposed using Fourier harmonic analysis into spherical power, regular astigmatism, and irregular astigmatism (asymmetry and higher order irregularity). RESULTS: On average, spherical power significantly increased after suture removal (p < 0.0001, paired t test), and regular astigmatism did not change significantly (p = 0.247). Suture removal significantly decreased asymmetry (p < 0.0001) and higher order irregularity (p < 0.0001). CONCLUSIONS: Suture removal after penetrating keratoplasty induces myopic shift and reduction in irregular astigmatism of the cornea. PMID- 11917173 TI - The stamler lid splint: a new short-term technique for achieving therapeutic ptosis. AB - PURPOSE: To provide an introduction to a new device, the Stamler lid splint (SLS), a disposable appliance designed to create a temporary total therapeutic ptosis. We also describe the indications for its use and analyze its limitations. METHODS: The utility of the SLS in effecting complete ptosis was examined in 33 patients. Outcome parameters examined were mean duration of (induced) ptosis, success (and failure) rate, and the incidence of side effects. RESULTS: The most common indications for using the SLS were lid closure abnormalities (n = 6) and epithelial defects after keratoplasty (n = 5). The induced ptosis lasted for a mean of 3.3 days (range, 1.5-6). The success rate of the SLS in achieving a total ptosis was 90.9%. Failure was attributed to the presence of atypical lid or orbital anatomy. No patient developed a complication specifically related to the SLS itself. CONCLUSIONS: The SLS is an inexpensive, quick, and technically straightforward means of treating a variety of ocular surface disorders in compliant patients with normal lid and orbital anatomy. The SLS is best suited to patients who require short-term therapy only. PMID- 11917174 TI - In vivo confocal microscopy after herpes keratitis. AB - PURPOSE: To describe the confocal microscopic findings, with special reference to corneal subbasal nerves, after herpes simplex virus (HSV) keratitis. METHODS: In this study, 16 HSV eyes and 14 contralateral eyes of 16 patients, diagnosed with unilateral HSV keratitis 1-12 months earlier by the presence of dendritic corneal ulceration or microbiologic confirmation, were examined by in vivo confocal microscopy for evaluation of corneal morphology. RESULTS: Herpes simplex virus eyes: In 2 eyes the surface epithelial cells appeared large, and no abnormalities were observed in the basal epithelial cells. In 2 eyes subbasal nerve fiber bundles were completely absent, in 3 eyes there was a reduced number of long nerve fiber bundles, and in 11 eyes the subbasal nerve plexus appeared normal. In 10 corneas, highly reflective dendritic structures were found at the level of the basal epithelial cells. Frequently these structures were found in the vicinity of stromal fibrosis. Areas with increased abnormal extracellular matrix were found in 11 eyes. Stromal nerves were not visualized in all corneas, but appeared normal when observed. Contralateral eyes: No abnormalities were observed in the epithelium. All corneas presented with a normal subbasal nerve plexus, but in 2 eyes dendritic particles were observed. Three corneas presented with activated keratocytes and increased amounts of abnormal extracellular matrix. CONCLUSIONS: When visualized by confocal microscopy, the subbasal nerve plexus appears relatively unaffected in cases with resolved HSV keratitis. Unidentified dendritic structures, presumably Langerhans cells, are frequently seen at the level of the basal epithelium in corneas with a history of herpetic disease. PMID- 11917175 TI - VIP stimulation of cAMP production in corneal endothelial cells in tissue and organ cultures. AB - PURPOSE: To demonstrate that vasoactive intestinal peptide (VIP), an immunosuppressive factor found in the aqueous humor, is a modulator of the corneal endothelium (CE) stimulating its intracellular cAMP production. METHODS: Rabbit CE cells in cell culture and CE cells in cornea cup organ cultures established from bovine and human donor eyes were treated with VIP at varying concentrations (0, 10(-11)-10(-6) mol/L) for a constant time (4 minutes) or varying times (1, 3.25, 10, 15 minutes) at a constant concentration (1 x 10(-6) mol/L). Intracellular cAMP was extracted and its concentrations were determined by radioimmunoassay. Agonists that are known to modulate the intracellular cAMP concentrations of target cells were allowed to react with cultured rabbit CE cells at 1 x 10(-6) mol/L for 4 minutes. RESULTS: Vasoactive intestinal peptide stimulated the intracellular cAMP production in CE cells in a dose- and time dependent manner. At concentrations lower than 10(-9) mol/L, VIP showed little effect. Treatment with 10(-8), 10(-7), and 10(-6) mol/L VIP for 4 minutes, however, increased the intracellular cAMP by 5.7-, 12.3-, and 9.5-fold, respectively, compared with the basal level in rabbit CE cell cultures, and by 19.5-, 38.7-, and 23.3-fold, respectively, in CE cells in bovine cornea cups. The effect of VIP was confirmed in two pairs of donor human corneas in which an average of 2.7-fold stimulation by 5 x 10(-7) mol/L was observed. Treatment of rabbit CE cells with 1 x 10(-6) mol//L VIP for 1 to 15 minutes elevated the intracellular cAMP level by six- to 69-fold. Among the agonists tested, alpha melanocyte-stimulating hormone and glucagon were not effective, whereas l isoproterenol and prostaglandin E1 were capable of stimulating the intracellular cAMP levels in rabbit CE cells. CONCLUSIONS: The current study demonstrated that VIP stimulated cAMP production in CE cells, similar to that shown previously in trabecular meshwork and nonpigmented ciliary epithelial cells. Tissues bathed in the aqueous humor are thus responsive to VIP modulation. PMID- 11917176 TI - Amniotic membrane graft for the management of scleral melting and corneal perforation in recalcitrant infectious scleral and corneoscleral ulcers. AB - PURPOSE: To study the efficacy and safety of using cryopreserved human amniotic membrane (AM) graft as a patch graft to reduce stromal melting and promote reepithelialization in extensive infectious scleral and corneoscleral ulcers. METHODS: Four cases of infectious scleral ulcers with persistent scleral melting and no sign of reepithelialization and three cases of corneoscleral ulcers with corneal perforation were studied. All patients had previously undergone pterygium excision, and infections were caused by Pseudomonas (n = 4), fungi (n = 2), and atypical Mycobacterium (n = 1). The area of limbus involved ranged from 3 to 9 (mean, 4.7) o'clock positions. Repeated debridements were performed, the causative microorganisms were identified, and the appropriate topical and systemic antibiotics were given to all patients before AM grafting. Postoperatively, the speed of reepithelialization, changes in the severity of scleral melting and inflammation, recurrence of infection, and visual acuity were documented. RESULTS: Melting and inflammation at the lesion site decreased after AM grafting. Reepithelialization of the scleral lesions was complete at an average 15.7 +/- 8.7 days (range, 5-31) postoperatively. Focal melting of the AM graft occurred in two cases, and in one case, it was necessary to perform further corneoscleral graft. No recurrent infection was encountered, but sterile abscess occurred in three cases that were located away from the original lesion. Useful vision above 20/400 was maintained in all patients at the end of follow-up. CONCLUSION: The AM graft is effective in promoting conjunctival reepithelialization and reducing scleral melting and inflammation and can be considered as an alternative biomaterial to improve wound healing in scleral and corneoscleral ulcerations. PMID- 11917177 TI - A study of glaucoma after penetrating keratoplasty. AB - PURPOSE: To identify the incidence and risk factors associated with the development of glaucoma after penetrating keratoplasty. METHODS: A retrospective study was carried out in 228 patients who underwent penetrating keratoplasty from January 1995 to January 2000 at the Cornea Section of the Ophthalmology Service at the School of Medicine, Federal University of Uberlandia MG, Brazil. Follow-up time ranged from 4 to 60 months (mean, 17.14 +/- 13.65). RESULTS: Two hundred twenty-eight patients undergoing penetrating keratoplasty were evaluated and 49 (21.5%) developed glaucoma. Risk factors for developing glaucoma were bullous keratopathy [relative risk (RR) = 2.1774), herpesvirus (RR = 1.8979), and trauma (RR = 1.0575). Mean intraocular pressure before transplantation was 15.26 +/- 5.37 mm Hg. Intraocular pressure after transplantation ranged from 22 to 58 mm Hg, with a mean value of 30.04 +/- 7.06 mm Hg. Thirty-six (73.5%) patients had medical treatment, 10 (20.4%) had medical and surgical treatment, and three (6.1%) had surgical treatment. Final intraocular pressure ranged from 4 to 34 mm Hg (mean, 16.79 +/- 5.36). CONCLUSION: The incidence of glaucoma after penetrating keratoplasty is high, and the risk factors include aphakic/pseudophakic bullous keratopathy, herpes simplex virus, and trauma. PMID- 11917178 TI - Topical corticosteroids of limited potency promote adenovirus replication in the Ad5/NZW rabbit ocular model. AB - PURPOSE: To determine the effect of topical therapy with several corticosteroids with limited potency on viral clearance in the adenovirus type 5 (Ad5) rabbit ocular model. METHODS: Sixty rabbits were inoculated in both eyes with Ad5. On the first day, the rabbits were equally divided into four topical treatment groups: 0.12% prednisolone acetate (PA), 0.1% fluorometholone (FM), 1% rimexolone (RMX), and control. Treatment was administered four times daily, in both eyes, for 3 days. All eyes were cultured for virus on days 0, 1, 3, 4, 5, 7, 9, 11, 14, 16, 18, and 21. RESULTS: Compared with the control group, treatment with PA, FM, and RMX significantly increased the number of Ad5 positive eye cultures from days 7-21. Fluorometholone and RMX prolonged the duration of Ad5 shedding, and FM increased the mean combined Ad5 titer from days 1-5 and 7-21. CONCLUSIONS: Treatment of an experimental ocular adenovirus infection with PA, FM, and RMX for 3 days significantly enhanced adenovirus replication compared with the control group. Short-term treatment of EKC with several commercially available topical corticosteroids with limited potency may offer symptomatic relief, but may also delay viral clearance and promote office and community epidemics. PMID- 11917179 TI - Effect of gefarnate on the ocular surface in squirrel monkeys. AB - PURPOSE: To investigate the ability of gefarnate (geranyl farnesylacetate) to stimulate goblet cell function in the primate eye after a mild alkali injury of the tarsal conjunctiva. METHODS: A bilateral injury was created on the conjunctival surface of the lower eye lid of squirrel monkeys by means of a 30 second application of a 4-mm diameter piece of filter paper wetted with 0.5% NaOH. Gefarnate drops (1%) were administered to one eye of each monkey and vehicle alone in the contralateral eye six times a day, 5 days a week for 4 weeks. Slit-lamp biomicroscopy, impression cytology staining of the ocular surface, fluorescein and rose bengal staining, and Western blot for mucin were performed before injury and weekly thereafter. Light microscopy was used to evaluate the lower conjunctiva. RESULTS: Topical application of gefarnate was not associated with any adverse ocular surface effects. Goblet cell repopulation after injury was significantly greater in the gefarnate-treated eyes compared with the vehicle-treated eyes. In the gefarnate-treated eyes, tear mucin content was significantly greater at 1 week after injury. Fluorescein staining was significantly reduced at 3 weeks after injury, and rose bengal staining was significantly reduced in the area of the wound at 2 weeks in the gefarnate treated eyes compared with the vehicle-treated eyes; at other times, conjunctival staining in the two groups of eyes was not significantly different. CONCLUSIONS: Gefarnate promotes goblet cell repopulation and increases mucin production after a conjunctival injury. No adverse affects of the treatment were found. Thus, this agent may be useful in conditions that diminish goblet cell function. PMID- 11917180 TI - Corneal endothelial damage after trabeculectomy with mitomycin C in two patients with glaucoma with cornea guttata. AB - PURPOSE: To report two patients with glaucoma who exhibited severe damage to the corneal endothelium after a trabeculectomy with mitomycin C (MMC). METHODS: This study includes clinical histories and specular microscopic pictures of the cases. RESULTS: Both patients were middle-aged women, underwent trabeculectomy with MMC, had moderate to severe cornea guttata preoperatively, and developed a shallow to flat anterior chamber, classified as grade 2 according to Spaeth early in the postoperative period. Stromal opacity caused by corneal edema associated with severe Descemet's membrane folds appeared within 2 to 5 days in both cases. The density of the corneal endothelium was decreased on specular microscopic examination. The severe corneal endothelial damage seen after the trabeculectomy with MMC was likely owing to a combination of the preexisting cornea guttata, the flat anterior chamber, and possibly the administration of MMC. CONCLUSION: Severe endothelial damage after trabeculectomy with MMC may occur in patients with glaucoma and associated cornea guttata. The use of tight sutures on the scleral flap or a modified operative method, nonpenetrating trabeculectomy, may be effective in preventing a shallow to flat anterior chamber postoperatively. PMID- 11917181 TI - Surgically induced necrotizing scleritis after pterygium excision and conjunctival autograft. AB - PURPOSE: To report a case of surgically induced necrotizing scleritis (SINS) after pterygium excision and conjunctival autograft. METHODS: A 55-year-old man presented 2 weeks after excision of primary pterygium with conjunctival autograft in the right eye with severe pain in that eye. He had undergone cataract surgery in that eye 8 months before. The graft was pale and white. The underlying sclera was white and avascular. There was marked inflammation adjacent to the site of surgery and graft. A scraping from the graft surface revealed no organisms in smears, and culture revealed no growth. The conjunctival graft was removed. Amniotic membrane transplantation was performed. After surgery, the amniotic membrane was avascular and pale. Systemic steroid therapy was initiated. RESULTS: Three days after initiation of systemic steroid therapy, the graft became vascularized. Over the next 10 days, the eye quietened, conjunctival and scleral inflammation lessened, and the graft was well accepted. CONCLUSIONS: Surgically induced necrotizing scleritis is a rare complication of pterygium excision with conjunctival autograft. A pale graft with underlying avascular sclera and surrounding inflammation should arouse suspicion of this complication. PMID- 11917182 TI - Differences in amyloid deposition in primary and recurrent corneal lattice dystrophy type 1. AB - PURPOSE: To report the histopathology of a case of recurrent corneal lattice dystrophy showing altered distribution of the corneal deposits in the recurrent disease compared with the original. METHODS: Clinical details and histopathology of the primary and repeat corneal grafts are reported. RESULTS: A woman originally presented at age 28 years with reduced visual acuity and classic corneal lattice lines in both corneas and underwent bilateral corneal grafts. Recurrent disease was detected 20 years later as anterior haze and various-sized subepithelial opacities but no stromal lattice lines. Histology of the original corneas demonstrated amyloid deposits throughout the corneal stroma, typical of corneal lattice dystrophy. In the repeat grafts, amyloid deposits were confined to the basement membrane region of the anterior cornea and were almost entirely absent from the stroma of the cornea. CONCLUSION: Recurrence of corneal lattice dystrophy is widely recognized to occur, but the pathology of the recurrent disease is not well documented in the literature. This case report highlights that there may be a difference in the distribution of the deposits when the disease recurs. We postulate that the reason for this difference may be that donor keratocytes survive long enough in the transplanted cornea to prevent build up of abnormal keratoepithelin, the product of the mutated gene in type 1 corneal lattice dystrophy. By contrast, the epithelium, being replaced by host epithelium shortly after grafting, is still producing abnormal protein. The differences in the pattern of deposits may have important clinical implications, particularly regarding treatment modalities in recurrent disease. PMID- 11917184 TI - Subepithelial corneal amyloid deposits in a case of congenital glaucoma: a case report. AB - PURPOSE: To report a rare association of subepithelial amyloid deposits in a long standing case of congenital glaucoma. METHODS: A 5-year-old girl was brought to the emergency service with complaints of bleeding from the left eye after injury to the eye with the door handle. Parents gave history of enlarged black portion of the eye since birth with increasing white opacity from the age of 6 months. On examination under general anesthesia, the left eye showed a limbal tear extending from 7 to 3.30 o'clock position and uveal prolapse. The right eye showed corneal edema, megalocornea, central white raised plaque, and high intraocular pressure. A diagnosis of bilateral congenital glaucoma with open globe injury, left eye, corneal scarring with degeneration, right eye was made and was advised enucleation. The enucleated eye was submitted for routine histologic examination. At a 3-year follow-up, there was an increase in the corneal scarring, with no significant improvement in vision. The child refused keratoplasty and was referred to low vision centre for visual rehabilitation. RESULTS: The enucleated eye was distorted and filled with blood. The cornea showed confluent pink homogenous wavy deposits in the subepithelial region. These deposits appeared brick red with Congo Red stain and showed apple green birefringence when viewed under polarized filters, confirming the amyloid nature of the deposits. There was loss of ganglion cell layer in the detached retina. CONCLUSION: The subepithelial amyloid deposits, presumably bilateral, in advanced case of congenital glaucoma were possibly secondary to the long-standing edema, scarring and vascularization. These secondary changes should be kept in mind as they could contribute to further deterioration in vision in advanced cases. PMID- 11917183 TI - Preseptal cellulitis caused by trichophyton (ringworm). AB - A 10-year-old boy with a past medical history significant for chicken pox at 7 years of age was referred to our eye center by an outside ophthalmologist for a 15-day history of worsening right-sided preseptal cellulitis. The patient reported photophobia, pruritus, and pain in the eyelid region. There appeared to be vesicular lesions on the eyelids. Empiric therapy with oral antibacterial and antiviral medications failed to resolve the preseptal cellulitis. Lid cultures revealed coagulase negative Staphylococcus, Streptococcal viridans, and a Trichophyton species of fungus. The infection was successfully treated with two courses of oral itraconazole. This is the first case of preseptal cellulitis caused by this dermatophyte that we could find in the literature. PMID- 11917185 TI - Case reports of three atypical infiltrative keratitis events with high DK soft contact lens wear. AB - PURPOSE: We report three atypical infiltrative keratitis events in patients that had successfully worn highly oxygen permeable (Dk) soft contact lenses on an extended wear (EW) schedule for at least 15 months. These cases highlight the role of patient/practitioner education, patient compliance, examination, and appropriate referrals that are required to reduce the risk of complications during lens wear. METHODS AND RESULTS: Each patient had been wearing high Dk soft lenses on an EW schedule with monthly replacement for at least 15 months and presented with severe pain, redness, and photophobia. The events were characterized by focal infiltrate(s) with an overlying epithelial defect in the superior periphery to mid-periphery of the cornea and extensive diffuse infiltration. CONCLUSIONS: The signs and symptoms of each case were more severe than is typically associated with contact lens-related infiltrative keratitis. Because of the slow progression of signs and relatively fast resolution, the cases were not found to be microbial keratitis. Although high Dk soft contact lenses overcome hypoxia-associated complications associated with extended wear, patients and practitioners must be vigilant as adverse events can still occur and in rare instances can be severe. Education and patient compliance are a crucial part of successful management of patients on EW schedules. Constant reinforcement is necessary to ensure patient compliance. PMID- 11917186 TI - Sutureless, posterior lamellar keratoplasty: a case report of a modified technique. AB - PURPOSE: To describe a technique for sutureless, posterior lamellar keratoplasty. METHODS: The procedure was performed for a case of pseudophakic bullous keratopathy. Through a 5.0-mm, self-sealing scleral tunnel incision, a stromal pocket was dissected across the cornea, just above Descemet's membrane. An 8.5-mm diameter posterior lamellar disc, consisting of posterior stroma, Descemet's membrane, and endothelium, was transplanted without suture fixation. RESULTS: One week after surgery, the best spectacle corrected visual acuity (BSCVA) was 0.8 (20/25), with S -1.5 and C -1.0 x 85 degrees. After 1 year, the posterior transplant was clear and in position, and the BSCVA was 0.8 with S -1.5 and C 1.75 x 80 degrees. Pachymetry measured 0.60 mm. Endothelial cell counts averaged 1390 cells/mm2. CONCLUSION: Sutureless, posterior lamellar keratoplasty may be an effective new surgical approach for managing corneal endothelial disorders. PMID- 11917188 TI - Keratoconus associated with cone-rod dystrophy: a case report. AB - A 31-year-old man with bilateral keratoconus associated with apical corneal scarring underwent uneventful penetrating keratoplasty in his left eye. Postoperatively his best-corrected visual acuity did not improve beyond 20/120. Examination of the fundus revealed features suggestive of bull's eye maculopathy. On performing an electroretinogram study of both eyes, grossly delayed implicit time with reduced amplitudes of the rod response and extinguished cone waveforms were noted, indicating cone-rod dystrophy. Color blindness was also noted on testing with Ishihara's pseudoisochromatic plates. Genetic counseling showed this to be an isolated defect with negative family history. Preoperative electrophysiologic studies may have a prognostic role in these cases. PMID- 11917187 TI - Ocular lesions arising after stings by hymenopteran insects. AB - PURPOSE: To describe the ocular lesions that occur after stings from hymenopteran insects. METHODS: We examined the ocular alterations in five patients who suffered ocular trauma from hymenopteran insect stings. RESULTS: In 4 cases where the insect was identified as a wasp, all the patients presented with persistent corneal decompensation and two presented with total cataract, requiring surgical treatment. In the case of trauma by a bee sting, the patient presented with corneal edema and an inflammatory reaction of the anterior chamber with total regression of these alterations after clinical treatment. CONCLUSION: Although a rare occurrence, ocular trauma caused by hymenopteran insects can result in severe ocular alterations in humans. PMID- 11917189 TI - Isolation of a vesicular virus belonging to the family rhabdoviridae from the aqueous humor of a patient with bilateral corneal endotheliitis. AB - PURPOSE: To report bilateral corneal endotheliitis caused by a vesicular virus (family Rhabdoviridae). METHODS: Case report of a 49-year-old man with a complaint of sudden onset of decreased vision in both eyes had diffuse corneal stromal edema with extensive folds in Descemet's membrane and was diagnosed as having bilateral viral endotheliitis. Virologic investigations were performed using aqueous humor from the right eye. RESULTS: An ether- and chloroform sensitive cytopathic agent was isolated in Vero and BHK-21 cell lines from the aqueous humor. It was identified as a vesicular virus belonging to the family Rhabdoviridae by electron microscopy. Neutralizing antibody was demonstrated at a titer greater than 1 in 4,096 dilutions in the convalescent serum. Neurologic complications included loss of hearing and postinfectious polyradiculopathy affecting both lower limbs. Best-corrected visual acuity was 20/120 OD and 20/20 OS. Six months later, he developed glaucoma in the right eye. Trabeculectomy with intraoperative application of 5-fluorouracil was performed. CONCLUSION: This is the first report of bilateral endotheliitis caused by a vesicular virus and confirmed by virus isolation from the aqueous humor of the affected eye. PMID- 11917190 TI - Response to "penetrating keratoplasty...". PMID- 11917191 TI - Utility of Etest in choosing appropriate agents to treat fungal keratitis. PMID- 11917193 TI - The prevalence of impaired left ventricular diastolic filling is related to the extent of coronary atherosclerosis in patients with stable coronary artery disease. AB - AIMS: The relation between abnormal left ventricular (LV) diastolic filling and the extent of coronary atherosclerosis per se has not been described. We aimed to investigate the prevalence of impaired LV diastolic filling in patients with stable coronary artery disease (CAD) and its relationship to the number and location of coronary lesions visualized at coronary angiography. METHODS AND RESULTS: In 170 consecutive patients with stable CAD and an abnormal coronary angiogram we assessed LV diastolic filling by Doppler evaluation of the transmitral early to atrial peak flow velocity (E/A) and the systolic to diastolic ratio of the pulmonary venous peak inflow to the left atrium (S/D). Abnormal diastolic filling was defined as E/A < or =0.75, or E/A >1.0 combined with S/D < or =1.0, and was present in 41% of the patients. In patients with one , two- and three-vessel disease the prevalence of impaired diastolic filling was 27, 30 and 49%, respectively (P = 0.026). In multiple logistic regression analysis diastolic filling was independently correlated with the number of stenotic coronary vessel areas. CONCLUSION: In patients with stable angiographically verified CAD, the prevalence of impaired diastolic filling was 41%. The prevalence increased with an increasing number of stenotic coronary artery areas independent of other variables tested, including prior myocardial infarction, LV systolic function and mitral regurgitation. PMID- 11917195 TI - Time course and extent of collateral channel recruitment during coronary angioplasty. AB - OBJECTIVES: To assess the extent and timing of recruitment of collateral channels during coronary angioplasty in patients without spontaneous collaterals at diagnostic angiography. SETTING: The extent of collateral channel recruitment during coronary angioplasty is variable and its contribution to myocardial protection is not well established. The functional significance of collaterals recruited during balloon occlusion remains in question. PATIENTS: Collateral channels were assessed in 16 patients by contralateral injection at 30, 60 and 90 s into each of four 90 s inflations and by a 0.014 " Doppler guide wire distal to the lesion. RESULTS: Angiographic collateral recruitment was evident in 11 out of 16 patients (71%), but in only four (24%) by intracoronary Doppler. Grade I collaterals were present in seven patients, grade II in three and grade III in two. Collaterals were evident angiographically by 30 s in 10 out of 11 patients, with no progressive recruitment during subsequent inflations. In the four patients with Doppler evidence of collateral flow there were no differences in any flow velocity parameters with successive inflations. There was no difference in either maximum ST segment shift or time to 2 mm ST segment elevation between successive inflations. CONCLUSIONS: Collateral channel recruitment is variable between patients and appears maximal early in the first inflation. The lack of incremental recruitment of collaterals together with low or absent evidence of flow by Doppler wire suggests that these channels do not make a major contribution to myocardial protection in this setting. PMID- 11917194 TI - Association of lipids, lipoproteins, apolipoproteins and paraoxonase enzyme activity with premature coronary artery disease. AB - BACKGROUND AND DESIGN: The association of serum apolipoprotein (apo) A-I and apo B concentrations and paraoxonase (PON) enzyme activity with angiographically determined coronary artery disease (CAD) was investigated in Iranian non-diabetic patients with premature CAD and control participants in a sex- and age-matched case-control study. METHODS: The study population consisted of 59 non-diabetic patients with premature CAD and 55 CAD control participants. Premature CAD was defined as the presence of angiographically proven coronary stenosis (> or =50% involvement) in men and women younger than 55 and 65 years, respectively. Apolipoprotein concentrations were measured by immunoturbidometric assay and paraoxonase/arylesterase activities by spectrophotometric assay of p nitrophenol/phenol production following addition of paraoxon/phenylacetate to serum. RESULTS: In CAD patients, increased concentrations of total cholesterol (215 +/- 43 compared with 193 +/- 43, P < 0.001), low-density lipoprotein cholesterol (137 +/- 46 compared with 116 +/- 39, P < 0.05) and apo B (102 +/- 24 compared with 84 +/- 17, P < 0.001) and a decreased ratio of apo A-I/apo B (1.7 +/- 0.4 compared with 2.0 +/- 0.6, P < 0.001) were observed compared to the control group. Other study variables were not significantly different between the two groups. On multiple logistic regression analysis, the only marker for discrimination between the CAD+ group and the CAD- control group was apo B level. CONCLUSIONS: In Iranian non-diabetic patients with premature CAD, the concentration of apo B is a better marker than traditional lipids in discriminating between CAD+ and CAD- patients. The lack of significant difference in PON activity between CAD patients and control participants supports the concept of interethnic variability in PON activity and gene polymorphism observed in other studies. PMID- 11917196 TI - Cardiocytic apoptosis and capillary endothelial swelling as morphological evidence of myocardial ischemia in ventricular biopsies from patients with angina and normal coronary arteriograms. AB - BACKGROUND: The cause of chest pain in patients with normal coronary arteriograms (CAG) remains poorly understood. METHODS AND RESULTS: Left ventricular endomyocardial biopsies from 11 anginal patients with normal CAG and normal left ventriculograms and from seven anginal patients with coronary stenosis were studied by light and electron microscopy. Biopsies from seven non-anginal patients (non-ischemic electrocardiogram abnormalities but no evident heart or systemic diseases) served as controls. In anginal patients with normal CAG, both cardiocytic diameter (17.2 +/- 5.5 microm) and interstitial space percentage area (37.6 +/- 14.9%) were significantly larger than those (13.7 +/- 0.9 microm, 14.9 +/- 2.9%) in control participants. Some cardiocytic nuclei (1.9% of 2000 randomly selected nuclei) exhibited DNA degradation by in-situ nick-end labeling. Electron microscopy revealed cardiocytic nuclei with distinct apoptotic ultrastructures (2.8% of 200 nuclei), phagocytic degradation of cardiocytic cytoplasm, and capillary endothelial swelling (7.1% of 200 capillary transverse sections). No significant infiltration of inflammatory cells was seen. In anginal patients with coronary narrowing (cardiocytic diameter, 16.8 +/- 1.1 microm; interstitial space, 20.1 +/- 5.8%; DNA degraded nuclei, 1.3%), there were however no apoptotic cardiocytic nuclei or cytoplasm and less capillary endothelial swelling (1.6%) in ultrastructure. CONCLUSIONS: In biopsies from anginal patients with normal CAG, the presence of cardiocytic hypertrophy and replacement fibrosis are both abnormal. Cardiocytic apoptosis and capillary endothelial swelling, found by others as characteristic of experimental myocardial reperfusion injury, are evident. This supports the possibility of myocardial transient ischemia and reperfusion injury in patients with angina and normal CAG. PMID- 11917197 TI - Characteristics and long-term outcome of patients with acute chest pain or other symptoms raising suspicion of acute myocardial infarction in relation to whether they were hospitalized or directly discharged from the emergency department. AB - AIM: To describe the characteristics and outcome of patients who came to the emergency department due to chest pain or other symptoms raising suspicion of acute myocardial infarction (AMI) in relation to whether they were hospitalized or directly discharged from the emergency department. METHODS: All patients arriving to the emergency department in one single hospital due to chest pain or other symptoms raising suspicion of AMI during a period of 21 months were followed for 10 years. RESULTS: In all, 5362 patients fulfilled the given criteria on 7157 occasions; 3381 (63%) were hospitalized and 1981 (37%) were directly discharged. Patients who were hospitalized were older and had a higher prevalence of previous cardiovascular diseases. The mortality during the subsequent 10 years was 52.1% among those hospitalized and 22.3% among those discharged (P < 0.0001). Risk indicators for death were similar in the two cohorts. However, many of these risk indicators including age, a history of myocardial infarction, angina pectoris, congestive heart failure, hypertension, initial degree of suspicion of AMI, a pathologic electrocardiogram on admission and a confirmed AMI as underlying etiology were more strongly associated with the prognosis among patients directly discharged than among those hospitalized. Ten (0.5%) of the patients who were directly discharged from the emergency department were found to have a diagnosis of confirmed or possible AMI, making up 1% of all patients given such a diagnosis. These patients had a 10-year mortality of 80.0% compared with 65.7% among patients with a confirmed or possible AMI who were hospitalized. CONCLUSION: Of patients who came to the emergency department with acute chest pain or other symptoms suggestive of AMI about a third were directly discharged. Their mortality during the subsequent 10 years was half that of patients hospitalized. Various risk indicators for death were more strongly associated with prognosis in the patients who were directly discharged from the emergency department compared to those hospitalized. Of all patients given a diagnosis of confirmed or possible AMI, 1% were discharged from the emergency department. Their long-term mortality was high, maybe even higher than among AMI patients hospitalized. PMID- 11917198 TI - The effect of type II diabetes mellitus on platelet aggregation in patients who underwent percutaneous transluminal coronary angioplasty. AB - BACKGROUND: In noninsulin-dependent type II diabetic (tIIDM) patients it was reported that ADP-induced platelet aggregation response was increased with decreased level of platelet guanylate cyclase. This study was therefore designed to examine the effects of tIIDM on collagen-induced, in-vitro platelet aggregation in percutaneous transluminal coronary angioplasty (PTCA) patients. METHODS: Twenty patients with tIIDM and 30 nondiabetic patients who had successful PTCA were included in the study. Platelet-rich plasma samples from the patients before and after PTCA were treated with in-vitro collagen and platelet aggregation waves were calculated via the turbidometric method of Born. The maximum amplitude (%) and the ratio of changes after PTCA in the study participants were measured by these waves and data were compared by student's t tests and nonparametric methods. The maximum amplitude of collagen-induced platelet aggregation before and after the procedure was also compared using variant analysis. RESULTS: The change in collagen-induced maximum amplitude of platelet aggregation in both wave 1 and wave 2 was significantly more (P < 0.001- P < 0.001) in the tIIDM group. The ratio of restenosis seen in the control coronary angiography made 6 months after intervention was found to be significantly more in the tIIDM group (P < 0.05). CONCLUSIONS: Collagen-induced platelet aggregation response was greater in patients with tIIDM than in nondiabetic patients. This makes us think that tIIDM patients could need more potent antiplatelet therapy before PTCA after the blood glucose levels have been regulated. PMID- 11917199 TI - Long-term prognostic value of dipyridamole echocardiography in vascular surgery: a large-scale multicenter study. AB - BACKGROUND: Late cardiac events after non-cardiac major vascular surgery are an important cause of morbidity and mortality. The aim of the present study was to assess the value of a preoperative dipyridamole echocardiography test (up to 0.84 mg/kg over 10 min) in predicting late cardiac events in survivors of major non cardiac vascular surgery. DESIGN: Large-scale, multicenter, prospective, observational study design. METHODS: Two hundred and seventy-six patients (mean age 66 +/- 9 years) were studied prior to vascular surgery by dipyridamole stress echocardiography in four different centres. All patients underwent preoperative clinical risk assessment according to the American Heart Association guidelines. All underwent dipyridamole stress echocardiography according to standard high dose protocol. RESULTS: No major complications occurred during dipyridamole stress echocardiography. Sixty-three patients (23%) had a positive test. Patients were followed up for a median of 20 months. Cardiac events occurred in 43 patients (16%): five deaths, 18 myocardial infarctions and 20 cases of unstable angina. The difference between wall-motion score index (WMSI) at rest and peak stress (delta WMSI), using multivariate analysis, was an independent predictor of late cardiac death. CONCLUSION: Dipyridamole stress echocardiography performed before major vascular surgery identifies patients at high risk for late cardiac events. Stress echocardiographic parameters outperformed clinical variables in the long-term risk stratification in this set of patients. PMID- 11917200 TI - Acute right ventricular infarction: clinical spectrum, results of reperfusion therapy and short-term prognosis. AB - BACKGROUND: The role of thrombolytic therapy (TT) and percutaneous coronary interventions (PCIs) in subgroups of patients with right ventricular infarction (RVI) has not been evaluated. METHODS AND RESULTS: We risk-stratified 302 patients with RVI into three subsets upon admission. Class A (n=197) comprised patients without right ventricular (RV) failure, Class B (n=69) with RV failure and Class C (n=36) with cardiogenic shock. All eligible patients in Class A or B received either PCI or TT. Patients in Class C eligible for reperfusion were treated with PCI. All patients were evaluated for in-hospital major adverse cardiac events and short-term mortality. There was a statistically significant difference in in-hospital mortality among the classes. Classes B and C were the strongest indicators of in-hospital mortality. By multivariate analysis TT or PCI did not reduce mortality in Classes A and B, but a clinically favorable trend in mortality reduction was documented: both methods decreased RV dysfunction in Class B (from 97% to 61% with TT and to 28% with PCI; P < 0.001) and PCI reduced the risk of mortality in Class C (89.5% compared with 58%; P < 0.05). CONCLUSIONS: Classification into types A, B or C allows the prediction of mortality. The use of TT or PCI suggests a clinical favorable trend in the reduction of mortality in Class A, either is beneficial in Class B for decreasing morbidity and PCI appears to be the most appropriate procedure for Class C since it reduced mortality. PMID- 11917201 TI - The effects of lipid-lowering therapy on low-density lipoprotein auto-antibodies: relationship with low-density lipoprotein oxidation and plasma total antioxidant status. AB - BACKGROUND: Oxidized low-density lipoprotein (Ox-LDL) is believed to play an important role in the progression of atherosclerosis. Oxidative modification of low-density lipoprotein (LDL) is a prerequisite for rapid accumulation of LDL in macrophages and for the formation of foam cells. Because of high antioxidant levels in plasma, LDL oxidation is suggested to occur mainly in the subendothelial space of the arterial wall, where there is the concomitant presence of large amounts of reactive oxygen species generated by endothelial cells and activated leukocytes. After Ox-LDL formation, antibodies against this form of LDL may occur. Auto-antibodies against Ox-LDL (AuAb-Ox-LDL) show directly in in-vivo LDL oxidation. Many studies have indicated that the amount of antibodies in serum is positively correlated to the rate of progression of atherosclerotic plaques. DESIGN AND METHODS: In this study the effect of lipid lowering therapy on the levels of AuAb-Ox-LDL in patients with dyslipidemia was determined using atorvastatin (10 mg/day), and the relationship between the antibodies and plasma total antioxidant status (TAS) and LDL oxidation capacity was also investigated. Serum levels of AuAb-Ox-LDL, lipids, lipoproteins, TAS and susceptibility of LDL to oxidation were determined using lag time in 44 patients with dyslipidemia (29 with hypercholesterolemia and 15 with mixed-type hyperlipidemia). RESULTS: After lipid-lowering therapy, serum levels of AuAb-Ox LDL were found to be significantly decreased, by 18.7%, while lag time and plasma TAS were increased (31.3% and 7.6% respectively) in patients with dyslipidemia. The percentage change in lag time was found to be negatively correlated to the percentage change in AuAb-Ox-LDL (r = -0.31, P < 0.05). The percentage change in lag time also showed a positive correlation with the percentage change in TAS (r = 0.58, P < 0.01). AuAb-Ox-LDL levels decreased by 21.7% in patients with hypercholesterolemia and by 12.6% in patients with mixed-type hyperlipidemia. Also AuAb-Ox-LDL levels in patients with hypercholesterolemia were higher than in those with mixed-type hyperlipidemia (367 +/- 294 compared with 300 +/- 176 mU/l). CONCLUSION: It was concluded that lipid-lowering therapy may contribute to the reduction in levels of AuAb-Ox-LDL and the increase in the antioxidant capacity of plasma LDL and TAS. It was also suggested that the measurement of antibodies against Ox-LDL during lipid-lowering therapy may be used as an important marker for representing in-vivo LDL oxidation and atherosclerotic processes. PMID- 11917202 TI - Vegetables, fruits, fibre and colorectal cancer. PMID- 11917203 TI - Primary malignant mucosal melanoma of the head and neck region: pooled analysis of 60 published cases from India and review of literature. AB - Malignant melanoma arising in the head and neck mucosa is a rare entity with incidence ranging from 2% to 10%. Because of the lack of data, the biological behaviour of these lesions still remains unpredictable and outcome dismal. We carried out a literature review for cases of mucosal melanoma of the head and neck reported from India and performed a pooled analysis on the available data. A total of 60 cases of head and neck melanomas were reported, of which 46 were in men. Palate and alveolus were the commonest sites. A total of 29 (48.3%) patients had regional node metastasis at presentation while five (12%) had distant metastasis. Three-year overall survival of 27.7% was observed. However, the disease-free survival rates at 3, 5 and 6 years were 39.4%, 39.4% and 13.1%, respectively. Metastasis at presentation and use of adjuvant radiotherapy were found to be the only significant predictors of survival. Malignant mucosal melanoma has aggressive biological behaviour and poor outcome. Radical surgery and adjuvant radiotherapy may provide a better local control and may help in improving survival. PMID- 11917204 TI - Tobacco smoking- and alcohol drinking-attributable cancer mortality in Germany. AB - The aim was to estimate the cancer mortality explained by tobacco smoking and alcohol risk drinking. The methods included (1) smoking rates from the largest population survey and alcohol drinking data from the German National Health Survey, and (2) vital statistics from Germany. The tobacco- and alcohol-related data were analysed using formulas for the estimation of attributable fractions. The results show that 19.7% of all malignant neoplasm deaths are explained by tobacco smoking alone, a further 0.2% by alcohol consumption alone and a further 5.6% by tobacco smoking and alcohol drinking. It is concluded that this overlap in the two substance use behaviours has to be taken into account when considering attributable mortality data. PMID- 11917205 TI - Prevention of N-methyl-N'-nitro-N-nitrosoguanidine and saturated sodium chloride induced gastric carcinogenesis in Wistar rats by lycopene. AB - We investigated "the "chemopreventive potential of lycopene against gastric carcinogenesis induced in male Wistar rats by N-methyl-N'-nitro-N nitrosoguanidine (MNNG) and saturated sodium chloride (S-NaCl). Administration of lycopene inhibited MNNG+S-NaCl-induced gastric carcinogenesis as revealed by the absence of carcinomas. Lipid peroxidation, reduced glutathione (GSH), glutathione peroxidase (GPx), glutathione S-transferase (GST) and glutathione reductase (GR) were used to monitor the chemopreventive potential of lycopene. The extent of lipid peroxidation was significantly lower, whereas GSH, GPx, GST and GR were markedly enhanced in the gastric mucosa of tumour-bearing animals. Our data suggest that lycopene may exert its inhibitory effects by modulating the oxidant and antioxidant status in the gastric mucosa. PMID- 11917206 TI - Active and passive smoking and the risk of stomach cancer, by subsite, in Canada. AB - This study assessed the influence of active and passive smoking on the risk of stomach cancer by subsite. Mailed questionnaires were used to obtain information on 1171 newly diagnosed histologically confirmed stomach cancer cases and 2207 population controls between 1994 and 1997 in eight Canadian provinces. Data were collected on socio-economic status, lifestyle and passive smoking status. Odds ratios (OR) and 95% confidence intervals (95% CI) were derived by logistic regression. Compared with those who had never smoked, there was strongly increased risk for ex- and current smokers among subjects with cardial stomach cancer. For men with cardial cancer, the adjusted ORs were 1.9 (95% CI 1.2-3.0) and 2.6 (95% CI 1.6-4.3) for ex-smokers and current smokers, respectively, with a similar pattern among women. Among men, the adjusted ORs were lower for subsites of stomach cancer other than cardia. These findings suggest that active and passive smoking may play an important role in the development of cardial stomach cancer. PMID- 11917207 TI - Protective effect of acetaminophen against colon cancer initiation effects of 3,2'-dimethyl-4-aminobiphenyl in rats. AB - A previous investigation demonstrated the anticarcinogenicity of acetaminophen (APAP) against colon carcinogenesis in rats induced by 3,2'-dimethyl-4 aminobiphenyl (DMAB). DMAB was selected as a structurally related surrogate for heterocyclic amines, formed during cooking of protein, which are believed to be involved in human colon cancer. The objective of the present study was to ascertain whether the early initiating effects of this colon carcinogen are inhibited by APAP. Six groups of male F344 rats were treated over a 6-week period as follows: (1) vehicle (corn oil) for 6 weeks; (2) APAP in the diet at 1000 ppm daily for 6 weeks; (3) 50 mg/kg DMAB by gavage once a week for the last 4 weeks; (4) 5 mg/kg DMAB as for (3); (5) 1000 ppm APAP for 6 weeks and 50 mg/kg DMAB for the last 4 weeks; and (6) 1000 ppm APAP and 5 mg/kg DMAB as for (5). Colonic tissue was within normal limits in the control and APAP groups. In the APAP only group, apical enterocytic hypertrophy and hyperaemia over the entire surface epithelium was present. In the high-dose DMAB group, in the lower third of the crypts, foci of enlarged glands with hypertrophic cells exhibiting karyomegaly and anisokaryosis (FHE) of 3+ degree of severity were evident in 100% of the animals. Also, there were increases in periglandular fibrocytes, matrix and mononuclear cells (PF). In the low-dose DMAB group both FHE and PF changes with the same degree of severity were reduced. In rats given the low dose of DMAB plus APAP, FHE and PF with the same degree of severity (3+) was absent. Both DMAB exposures increased significantly the replicating fraction of colonic enterocytes in an exposure-related fashion and the replicating fractions were significantly reduced by APAP. In 32P-postlabelling of colon, liver and urinary bladder DNA, high-dose DMAB produced 2-6 distinct dose-related spots reflecting DNA adducts. These spots were reduced or were no longer detectable in all three tissues when APAP was given 2 weeks before and during DMAB exposure. Using immunohistochemical detection of DMAB adducts in the colon, a dose-related colour intensity was present for both doses of DMAB. APAP reduced this by 94-fold. Thus, APAP produced a marked protective effect in colonic enterocytes against several parameters of neoplastic development by the carcinogen. PMID- 11917208 TI - Detection of nodular and superficial spreading melanoma with tumour thickness < or = 2.0 mm--an interview study. AB - Differences regarding signs and symptoms between nodular melanoma (NM) and superficial spreading melanoma (SSM) with a tumour thickness < or = 2.00 mm were studied. Reasons for seeking medical attention were also investigated. Semi structured interviews were conducted with 22 patients with NM diagnosed between 1994 and 1999 and 32 patients with SSM, matched to the NM patients with respect to age at diagnosis, gender and tumour thickness. NM were smaller in diameter, and 5 of 6 patients reporting a diameter <5 mm had NM. NM were more often new lesions than SSM, which more often had developed from pre-existing naevi. In most cases (61%), the melanoma was first detected by the patient and in 17% it was detected by the patient in combination with a family member. The most important reasons to seek medical attention were a change in the lesion or a symptom (65%), followed by encouragement from others to seek medical attention. Twenty-four patients (44%) had no symptoms of melanoma prior to diagnosis. Our results have important implications for prevention. It should be emphasized in public education that melanomas may have a diameter <6 mm and that family members have an important role in detection of melanoma and in motivating medical advice. PMID- 11917210 TI - Low serum prostate-specific antigen levels in elderly rural African men at very low risk of prostate cancer. PMID- 11917209 TI - Comparison of the chemopreventive potentials of melatonin and vitamin E plus selenium on 7,12-dimethylbenz[a]anthracene-induced inhibition of mouse liver antioxidant enzymes. AB - Chemoprevention is a rapidly growing area of oncology that is identifying agents with a potentially preventive role in cancer. In this study, it was our goal to compare the chemopreventive effects of vitamin E plus selenium, and melatonin. Forty female mice were divided into four equal groups. The first group served as control. The second group had i.p. injections of 7,12-dimethylbenz[a]anthracene (DMBA) (20 mg/kg body weight) in corn oil for 21 days. The third group had the same procedure of DMBA injections as the second group and received vitamin E + selenium (90 microg + 1.8 microg/day), simultaneously. The fourth group had DMBA injections and melatonin (4.2 mg/kg body weight), simultaneously. DMBA alone caused significant inhibition of hepatic glutathione peroxidase (GSHPx), catalase (CAT), and superoxide dismutase (SOD) in the second group. In the third group, vitamin E + selenium restored DMBA-induced GSHPx inhibition significantly whereas CAT and SOD inhibition remained essentially unchanged. In the fourth group, melatonin not only significantly decreased DMBA-induced GSHPx inhibition but also fully reversed CAT and SOD inhibitions caused by DMBA. We speculate that melatonin alone provides better chemoprevention against DMBA-induced oxidative stress than the vitamin E+selenium combination. PMID- 11917211 TI - Smelly science. PMID- 11917212 TI - Methodological aspects of investigating gene-nutrient interactions. PMID- 11917213 TI - Cooking methods, metabolic polymorphisms and colorectal cancer. PMID- 11917214 TI - Molecular markers of heterogeneity in colorectal cancers and adenomas. PMID- 11917215 TI - Studies on gene-nutrient interactions in the aetiology of colorectal neoplasia ('ECP-genuine' project): ethical and legal aspects. PMID- 11917216 TI - Sources of DNA for studying gene-nutrition interactions. PMID- 11917217 TI - Colorectal neoplasia and genetic polymorphisms associated with folate metabolism. PMID- 11917218 TI - Salpingitis, salpingoliths, and serous tumors of the ovaries: is there a connection? AB - We have observed luminal and mucosal calcifications frequently surrounded by a mantle of bland epithelium in the fallopian tubes ("salpingoliths") of women with serous tumors of the ovaries. These lesions resemble noninvasive peritoneal "implants" in women with advanced stage atypical proliferative serous tumors (APSTs) and micropapillary serous carcinomas (MPSCs). The presence of salpingitis and salpingoliths was prospectively evaluated in 358 women with a variety of nonneoplastic and neoplastic ovarian conditions and compared with 87 previously reported women with APSTs/MPSCs in an effort to determine whether these lesions were specifically associated with serous tumors. The frequency of chronic salpingitis among women without ovarian pathology was 27%, and the frequency of salpingoliths was 4%. Serous epithelial tumors (cystadenomas, APST/MPSC, and carcinomas) were significantly more often associated with chronic salpingitis (53%) and salpingoliths (32%) than all other cases with or without ovarian neoplasms (p<0.01). APSTs/MPSCs were associated with salpingoliths significantly more frequently than all other groups (p<0.001). For patients with APSTs/MPSCs, salpingoliths were found significantly more often in advanced stage (FIGO II and III) patients (51%) than stage I patients (24%) (p<0.01), but salpingitis, present in 60% of these patients, was not stage-dependent (p>0.05). Chronic salpingitis was identified in 66% of women with endometriosis, which was significantly more frequent than those with normal ovaries (27%) (p<0.001). In conclusion, fallopian tube abnormalities may be related to both the high frequency of infertility and the noninvasive peritoneal implants in women with APSTs/MPSCs. Whether the fallopian tubes with salpingoliths are the source of the peritoneal "implants," the recipient of implants, or are independent is unknown. In addition, the high frequency of salpingitis in women with endometriosis may be related to the mechanism of endometriosis-associated infertility. PMID- 11917219 TI - Eosinophils as a marker for invasion in vulvar squamous neoplastic lesions. AB - A study of eosinophils directed against vulvar neoplastic squamous epithelium was undertaken to determine whether there were thresholds per high-power field (hpf) or 10 hpf that were a marker for invasion. The presence of stromal and intraepithelial eosinophils in 33 cases of vulvar grade 3 squamous intraepithelial neoplasia (carcinoma in situ) (VIN 3) was compared with that in 38 cases of vulvar invasive carcinoma with any degree of invasion (ISC). In both incisional biopsy and excisional specimens, the presence of >3 eosinophils per high-power field (eos/hpf) and the presence of >or=5 eosinophils per 10 high power fields (eos/10 hpf) were both significantly associated with invasion, and the presence of >or=20 eos/hpf and/or >50 eos/10 hpf was limited to cases with invasion. The presence of eosinophils within the neoplastic squamous epithelium was also limited to cases with invasion. The author proposes: 1) eosinophil counts in vulvar incisional biopsy specimens of >3/hpf and/or >or=5/10 hpf warrant a note of caution that invasion may be present even when none is identified by conventional criteria; 2) eosinophil counts of >3/hpf and/or >or=5/10 hpf in excisional specimens should raise the suspicion of invasion in cases in which only VIN 3 is identified in the initial sections, and warrant additional sections and/or levels to search for invasion; 3) the above eosinophil counts provide supportive evidence for invasion in cases with equivocal invasion by conventional criteria; and 4) the presence of >or=20 eos/hpf and/or >50 eos/10 hpf, and the presence of intraepithelial eosinophils in conjunction with >3 eos/hpf and >or=5 eos/10 hpf is virtually diagnostic of invasion. PMID- 11917220 TI - Eosinophils as a marker for invasion in cervical squamous neoplastic lesions. AB - A study of eosinophils associated with cervical neoplastic squamous epithelium was undertaken to determine whether their presence is a marker for invasion. Forty cervical incisional biopsy specimens of high-grade squamous intraepithelial neoplasia (HSIL), 12 with an element of invasive carcinoma, and 2 of HSIL suspicious for invasion, and follow-up excisional specimens of 27 cases of HSIL and 6 of microinvasive and invasive carcinoma were reviewed. In both incisional biopsy and excisional specimens, the presence of >or=5 eosinophils/high-power fields (hpf) and >or=10 eosinophils/10 hpf were both highly significantly associated with invasion with a high degree of specificity and positive predictive value, whereas counts below these thresholds had a high negative predictive value. The authors propose: 1) eosinophil counts in cervical incisional biopsy specimens of >or=5/hpf and/or >or=10/10 hpf warrant a note of caution that invasion may be present even when none is identified in the specimen by conventional criteria; 2) eosinophil counts of >or=5/hpf and/or >or=10/10 hpf in excisional biopsy and hysterectomy specimens should raise the suspicion of invasion in cases in which only HSIL is identified in the initial sections, and warrant additional sections and/or levels to search for invasion; and 3) the above eosinophil counts may provide supportive evidence for invasion in cases with equivocal invasion by conventional criteria. PMID- 11917221 TI - Mitotic activity and apoptosis in endocervical glandular lesions. AB - To evaluate the significance of mitotic activity and apoptosis in the differential diagnosis of endocervical glandular lesions, we examined the frequency of mitoses and apoptosis in 89 endocervical glandular lesions from 78 patients, which consisted of benign reactive changes (7 cases), lobular or diffuse laminar endocervical glandular hyperplasia (4), microglandular hyperplasia (3), tunnel clusters (7), nabothian cysts (2), mesonephric remnants (3), tubal metaplasia (3), endocervical glandular dysplasias (including atypical tubal metaplasia) (EGD) (7), adenocarcinoma in situ (AIS) (31), microinvasive adenocarcinoma (7), frankly invasive adenocarcinoma (12), and minimal deviation adenocarcinoma (3). Mitotic index (MI; mitotic figures per 1000 cells) was significantly higher in AIS, microinvasive adenocarcinoma, and frankly invasive adenocarcinoma than any other lesions examined. Microinvasive adenocarcinoma showed the highest MI. Apoptosis was detected consistently and frequently in AIS, microinvasive adenocarcinoma, and frankly invasive adenocarcinoma. AIS showed the highest apoptotic index (AI; apoptoses per 1000 cells). Frequent apoptotic bodies and mitotic figures are a common feature of endocervical glandular malignancies (except for minimal deviation adenocarcinoma) and are an important feature that can facilitate their differentiation from benign and borderline lesions. High MI in microinvasive adenocarcinoma might aid the distinction of microinvasive adenocarcinoma from AIS. Although both MI and AI of EGD were between those of benign reactive changes and of AIS, MI and AI alone are not sufficient to differentiate EGD from benign reactive changes. MI and AI are not helpful in the differential diagnosis between minimal deviation adenocarcinoma and its benign mimics. PMID- 11917222 TI - Glassy cell carcinoma of the uterine cervix: histochemical, immunohistochemical, and molecular genetic observations. AB - Glassy cell carcinoma (GCC) of the uterine cervix is characterized by distinctive cytological features and an aggressive clinical course. Although this tumor has been usually considered a poorly differentiated variety of adenosquamous carcinoma (ASC), a clarification of the phenotype and histogenesis of GCC is still required. We examined three GCCs and four ASCs for histochemical and immunohistochemical phenotypes and molecular genetic status, comparing them with five nonkeratinizing squamous cell carcinomas and five endocervical-type mucinous adenocarcinomas. GCCs had a profile of cytokeratin expression similar to that of reserve cells or immature squamous cells of the cervix. In addition to squamous differentiation, GCCs sporadically produced intestinal-type mucin. HPV 18 was detected in two of three GCCs and two of four ASCs. GCCs may originate from multipotential stem or reserve cells that undergo early squamous differentiation. The presence of HPV 18 might stimulate biphasic squamous and glandular differentiation. PMID- 11917223 TI - Nodular histiocytic hyperplasia of the endometrium. AB - We describe the clinical and pathologic features of four cases of nodular histiocytic proliferation in the endometrium. We have been able to find only one brief reference to this lesion in the literature. The lesion in each case was a detached nodule composed of aggregates of histiocytes within a biopsy or curettage specimen. The constituent cells differed from foamy histiocytes of the endometrium in that they had either lobulated or ovoid, vesicular nuclei, distinctive cytoplasmic margins, and a moderate amount of amphophilic cytoplasm. Mitoses were frequent (up to 11 per 10 high-power fields) in one case but were absent in the remaining cases. On immunohistochemical staining, CD68 and lysozyme were strongly expressed in the cytoplasm. Neither estrogen receptor nor progesterone receptor was expressed in contrast to the background endometrium. The cells were also negative for S-100 and cytokeratin. Each patient's postcurettage course was uneventful. The cause of nodular histiocytic proliferation of the endometrium is currently unknown, although response to intracavitary debris has been suggested. The lesion should not be confused with a variety of reactive, inflammatory, or neoplastic conditions, such as xanthogranulomatous endometritis, malakoplakia, histiocytic granuloma, hormonal changes of the endometrial stroma, Langerhans' cell histiocytosis, morular metaplasia, extravillous trophoblast, or exaggerated placental site reaction. PMID- 11917224 TI - Expression of COX-2, Ki-67, cyclin D1, and P21 in endometrial endometrioid carcinomas. AB - COX-2, the isoform of cyclooxygenase inducible by cytokines, mitogens, and growth factors, appears to play an important role in inflammation and carcinogenesis. In the colon, COX-2 overexpression results in cell cycle alterations, and NSAIDs have proven effective in cancer chemoprevention. HNPCC (hereditary nonpolyposis colon cancer) is a clinically defined cancer susceptibility syndrome in which women are also at significantly increased risk for the development of endometrial carcinoma. The purpose of this study was to evaluate expression of COX-2 in benign and malignant endometrium in the context of other cell cycle and proliferation markers, including Ki-67, cyclin D1, and the cyclin-dependent kinase inhibitor, p21. Immunostains with COX-2, Ki-67, cyclin D1, and p21 antibodies were performed on formalin-fixed and paraffin-embedded tissue sections from 40 cases: 10 benign (5 atrophic and 5 proliferative) endometria, 6 hyperplasias (complex without atypia), and 24 endometrioid carcinomas (9 well, 4 moderately, and 11 poorly differentiated). Ki-67 was positive in all proliferative and neoplastic endometria. Cyclin D1 and p21 were both overexpressed in endometrial hyperplasia and endometrioid carcinomas. COX-2 was negative in the nonneoplastic endometrium, stained minimally in the well differentiated endometrioid carcinomas, and stained most strongly in the moderately and poorly differentiated endometrioid carcinomas. Because cyclin D1 may function as an oncogene, its effects may dominate the usual inhibitory effect of a rising p21. Alternatively, it has been shown that p21 can promote cell cycle function by stabilizing cell cycle complexes. The overexpression of COX-2 in poorly differentiated endometrioid carcinoma and lack of expression in hyperplasia and well-differentiated carcinoma suggests that in this form of cancer, COX-2 may play a role in tumor progression rather than tumor initiation. PMID- 11917226 TI - Tenascin expression in normal, hyperplastic, and neoplastic endometrium. AB - Tenascin (TN) is an extracellular matrix glycoprotein (ECM) that participates in embryogenesis and carcinogenesis. The aim of this study was to investigate immunohistochemically the expression of TN in the normal, hyperplastic, and neoplastic endometrium (endometrial adenocarcinoma). In the adenocarcinomas, the results were correlated with patient age, menopausal status, stage, grade, myometrial invasion, and vascular invasion. TN expression was studied in the following cases: proliferative endometrium (10 cases), early secretory endometrium (10), secretory endometrium (10), simple hyperplasia (15), complex hyperplasia (15), atypical hyperplasia (15), and endometrial adenocarcinomas (25). Staining of basal membranes and the cytoplasm of the stromal and epithelial cells was evaluated semiquantitatively. Positive staining was observed in the vascular and glandular basal membranes, stromal cells, and epithelial cells of proliferative, hyperplastic, and neoplastic endometrium. The difference in percentage of stained stromal cells between the neoplastic and the nonneoplastic (proliferative and hyperplastic) endometrium was significant (p<0.005). However, the percentage of stained epithelial cell area in hyperplasia was significantly higher than that of adenocarcinoma and functional endometrium (p<0.005). We conclude that TN is an extracellular matrix glycoprotein that plays a role in proliferation and possibly endometrial carcinogenesis. PMID- 11917225 TI - Dissociated expression of Bcl-2 and Ki-67 in endometrial lesions: diagnostic and histogenetic implications. AB - The objective of the present study was to analyze the expression of the proliferation marker, Ki-67, and the anti-apoptotic protein, bcl-2, in various endometrial lesions. Ki-67 and bcl-2 expressions were studied in 194 specimens of endometrial hyperplasia, polyps, carcinomas, and cyclic endometrium from a defined geographic area. Results were statistically analyzed with respect to marker expression, localization to the stromal or glandular component, and intraglandular topography. The lowest glandular Ki-67 expression was seen in secretory endometrium, in polyps, and in atypical hyperplasia. The Ki-67 score was significantly higher and less heterogeneous in endometrial carcinomas than in hyperplasia (p<0.001). Endometrial hyperplasia of all types was characterized by a markedly heterogeneous glandular expression of Ki-67. The glandular expression of bcl-2 was highest in proliferative endometrium and polyps. Bcl-2 expression was significantly lower in adenocarcinomas than in hyperplastic lesions (p=0.002). Ki-67 and bcl-2 expression showed a significant association in proliferative endometrium (p=0.003). Endometrial polyps demonstrated a unique pattern of very low expression of Ki-67 and high bcl-2 expression in both stroma and glands. Our findings indicate that an imbalance between proliferation and apoptosis may be an important factor in the development of different endometrial lesions, benign as well as malignant. The specific finding of inter- and intraglandular Ki-67 heterogeneity may be valuable as an adjunct to morphology in the differential diagnosis of endometrial hyperplasia. PMID- 11917227 TI - Immunohistochemical analysis of alphavbeta5 and alphavbeta6 integrins in the endometrium and endometriosis. AB - We performed a case-controlled study to study the immunoreactivity patterns and staining intensity of alphavbeta5 and alphavbeta6 integrins in ectopic and eutopic endometrium from women with endometriosis and compared them with those in normal control endometrium. Forty eutopic and ectopic samples from patients with endometriosis were compared with 12 control endometrial samples. Staining was evaluated using a computerized image analysis system by a blinded, independent observer. The immunoreactivity patterns for both alphavbeta5 and alphavbeta6 were very similar. In eutopic endometrium from women with or without endometriosis, the immunostaining intensity score for both integrins was stronger during the secretory than the proliferative phase. Immunoreactivity in endometriosis was greater than in eutopic endometrium. There were no significant menstrual cycle related staining differences in endometriosis. The percentage of blood vessels immunolabeled was greater in endometriosis than eutopic endometrium from endometriosis patients and greater in eutopic endometrium from these patients compared with control endometrium. Ectopic tissue from patients with stages I and II endometriosis showed significantly higher staining intensity than ectopic tissue from patients with stages III and IV disease. Determination of the specific functional consequences of the differences observed in alphavbeta5 and alphavbeta6 integrin immunostaining may provide an increased understanding regarding the pathogenesis of endometriosis. PMID- 11917228 TI - Extramedullary hematopoiesis in the endometrium. AB - Extramedullary hematopoiesis (EMH) in the endometrium is an extremely rare occurrence. Four of the eight previously reported cases were related to an underlying hematological disorder, although the remainder had no such relationship. We describe a case of endometrial EMH associated with retained products of conception after termination of pregnancy. Routinely and immunohistochemically stained slides revealed several collections of normoblasts and granulocytic precursors in the endometrium with synchronous chronic endometritis. Retained chorionic villi were also identified. The patient had no known history of a hematological disorder or systemic disease and no such abnormality was detected after detailed hematological work-up. Local effects of growth factors on circulating stem cells may play a pathogenetic role in this process, although an association with recent pregnancy in this case suggests that implantation of fetal hematopoietic elements from the fetus or yolk sac may be more likely. PMID- 11917229 TI - Leiomyosarcoma of the uterus with a florid intravascular component ("intravenous leiomyosarcomatosis"). AB - A leiomyosarcoma of the uterus in a 54-year-old woman exhibited striking involvement of large vessels of the myometrium and broad ligament on both gross and microscopic examination. The pattern of vascular involvement resembled that seen in intravenous leiomyomatosis. Imaging studies showed recurrent tumor within the inferior vena cava 3 months after hysterectomy. To our knowledge, this is the first reported such case in the literature, for which we propose the designation intravenous leiomyosarcomatosis of the uterus. PMID- 11917231 TI - Ceroid granuloma of the uterine cervix. AB - A case of ceroid granuloma of the uterine cervix in a 58-year-old woman is presented, the fourth such case in the literature. It was an incidental finding during a routine pelvic examination and appeared as an exophytic brownish lesion on the anterior wall of the uterine cervix. On histological examination it consisted of sheets of ceroid-rich histiocytes within the superficial cervical stroma. There was no obvious cause for the lesion. Ceroid granulomas of the female genital tract may be related to endometriosis, but the rarity of the cases precludes definite conclusions about the etiology and pathogenesis of this lesion. PMID- 11917230 TI - Co-existent carcinosarcoma and adenoid basal carcinoma of the uterine cervix and correlation with human papillomavirus infection. AB - A case of carcinosarcoma associated with an adenoid basal carcinoma of the uterine cervix in an 84-year-old woman is described. The tumor formed a pelvic mass, and total hysterectomy with bilateral salpingo-oophorectomy was performed. The tumor was 17 cm in maximal dimension, arose in the posterior wall of the uterine cervix, and had a solid and cystic, focally myxoid sectioned surface. The tumor was composed of keratinizing squamous cell carcinoma and a mostly nonspecific sarcomatous component with focal chondrosarcomatous differentiation. Multiple foci of adenoid basal carcinoma were observed in the adjacent cervical wall. The HPV L1 gene was detected by PCR in each of the carcinomatous and sarcomatous components of the carcinosarcoma and the adenoid basal carcinoma. To our knowledge, this is the fifth case of coexisting carcinosarcoma and adenoid basal carcinoma of the uterine cervix in the English language literature. PMID- 11917232 TI - Salivary gland-type basal cell adenocarcinoma of presumed bartholin's gland origin: a case report. AB - Salivary gland-type carcinomas arising in Bartholin's gland are rare neoplasms. We report the case of a 75-year-old female who presented with a vulvar tumor with morphological and immunophenotypical features identical to those of salivary gland basal cell adenocarcinomas. We believe that the tumor was most likely of Bartholin's gland origin, although no Bartholin's gland tissue was found adjacent to the neoplasm. This type of malignancy with myoepithelial differentiation has not been previously reported to arise in the Bartholin's gland. PMID- 11917233 TI - XII. Aurel Babes. PMID- 11917236 TI - Comparative analysis of HIV-1 viral load assays on subtype quantification: Bayer Versant HIV-1 RNA 3.0 versus Roche Amplicor HIV-1 Monitor version 1.5. AB - Quantification of HIV-1 subtypes is essential for appropriate clinical management. Whereas viral load assays were initially developed to accurately quantify subtype B, the recent worldwide spread of non-B subtypes and the introduction of treatment programs in regions with non-B subtypes have prompted adaptations of these assays. The Bayer Versant HIV-1 RNA 3.0 Assay (branched DNA [bDNA] 3.0) and the Roche Amplicor HIV-1 Monitor version 1.5 (Amplicor 1.5) assays are reported to quantify all subtypes in group M; however, evaluation of performance characteristics remains limited. In this study, we evaluated the accuracy and reliability of bDNA 3.0 and Amplicor 1.5 on multiple serially diluted viral isolates from HIV-1 group M, subtypes A through F. Testing was conducted on both assay systems in two independent laboratories. Comparative pansubtype quantification from regression analysis showed that quantification by bDNA 3.0 was approximately 0.3 log-fold lower than that by Amplicor 1.5. Comparative pansubtype accuracy analysis showed data points more closely distributed about their respective regression lines and thus showing greater reliability by bDNA 3.0 than by Amplicor 1.5. PMID- 11917235 TI - Transplacental genotoxicity of combined antiretroviral nucleoside analogue therapy in Erythrocebus patas monkeys. AB - Antiretroviral nucleoside analogue drugs are a major constituent of highly active antiretroviral therapy (HAART), the most advanced form of treatment for HIV-1 infection. Currently, HAART combinations that include zidovudine (ZDV) and lamivudine (3TC) are highly effective in preventing HIV-1 vertical transmission; most children are born with no evident adverse clinical effects. However, ZDV is a moderately strong transplacental carcinogen in mice, and potential long-term consequences of fetal exposure to most HAART combinations remain unknown. To model human transplacental ZDV and 3TC exposures, experiments were performed in Erythrocebus patas monkeys given human-equivalent drug exposure protocols. Pregnant monkeys were dosed with either no drug (n = 2), 40.0 mg ZDV/d (about 6 mg/kg body weight/d) for the last 50% (10 weeks) of gestation (n = 3), or with the same regimen of ZDV plus 24.0 mg 3TC/d (about 3.6 mg/kg body weight/d) for the last 20% (4 weeks) of gestation (n = 3). Multiple fetal organs were examined at term for DNA incorporation of ZDV and 3TC using two separate radioimmunoassays (RIAs). Values for ZDV-DNA incorporation were similar in fetuses exposed to ZDV alone and those exposed to ZDV plus 3TC. Values for 3TC-DNA in fetal organs were greater than or equal to values for ZDV-DNA, indicating that the total DNA damage sustained by fetuses exposed to both drugs was at least double that observed in fetuses exposed to ZDV alone. Telomere shortening, determined by Southern blot with a telomeric probe, was observed in most organs of the three animals exposed in utero to ZDV plus 3TC. No telomere shortening was evident in the unexposed fetuses, and occasional telomere shortening was found in fetuses exposed to ZDV alone. Overall, these studies demonstrate that monkey fetuses exposed in utero to the combination ZDV plus 3TC sustain a higher level of drug-DNA incorporation and show evidence of more telomere damage than monkey fetuses exposed to ZDV alone. PMID- 11917237 TI - Lack of hepatotoxicity associated with nonnucleoside reverse transcriptase inhibitors. AB - Nonnucleoside reverse transcriptase inhibitors (NNRTIs), particularly nevirapine, have been associated with hepatotoxicity. We performed a retrospective study to determine the incidence of NNRTI hepatotoxicity in a group of HIV-infected patients from a New York City practice. These patients are predominantly homosexual white males. We also analyzed the effect of coinfection with hepatitis B (HBV) or hepatitis C (HCV) virus. In total, 272 patients received NNRTIS: 40 (15%) received delavirdine, 91 (33%) received efavirenz, and 141 (52%) received nevirapine. Of the patients with known hepatitis status, 18 of 190 (9%) were coinfected with HBV, and 24 of 205 were coinfected (12%) with HCV. The overall rate of grade 3 to 4 elevations in aspartate aminotransferase (AST) or alanine aminotransferase (ALT) was 3 of 272 (1.1%) and did not differ significantly among the three NNRTIs. HBV or HCV was not associated with a significant increase in AST or ALT elevations. We conclude that NNRTIs are relatively free from hepatotoxicity in this population, despite the presence of coinfection with HBV or HCV. PMID- 11917239 TI - Placebo-controlled trial of cyclosporin-A in HIV-1 disease: implications for solid organ transplantation. AB - OBJECTIVE: Earlier open-label clinical trials have provided conflicting data on the effects of cyclosporin-A (CsA) on the clinical course and immune status of patients with HIV disease. With the prospects for wider use of CsA in the setting of solid organ transplantation in HIV-infected persons, data on the safety and immunologic activity of this agent are needed. We report here the results of a randomized, double-blind, placebo-controlled trial to assess the safety and immunologic activity of CsA administration in early HIV disease. METHODS: Twenty eight patients with confirmed HIV infection, CD4 cell counts greater than 500 x 106/L, and plasma HIV RNA >600 copies/mL were randomized to receive 2 mg/kg of CsA (Neoral) twice daily or identical placebo for 12 weeks. Subjects were stratified for the presence or absence of stable concomitant antiviral therapy. The primary end point was the effect of therapy on immune activation as assessed by the levels of soluble interleukin-2 receptors. Secondary end points included safety and effects of treatment on plasma HIV RNA, CD4 cell count, and other markers of immune activation and function. RESULTS: The low dose of CsA used in this study did not suppress immune activation or increase circulating CD4 cell counts. Delayed-type hypersensitivity responses were not affected; however, lymphocyte proliferative responses tended to decrease. CsA-treated patients experienced a small but significant rise in plasma HIV RNA levels. CONCLUSIONS: Low-dose CsA has no benefit in patients with stable early HIV disease, and its administration may be associated with an increase in plasma HIV RNA. The use of CsA in HIV-infected patients undergoing organ transplantation should be undertaken with caution PMID- 11917238 TI - Predictive value of immunologic and virologic markers after long or short duration of HIV-1 infection. AB - Laboratory markers that predict HIV-1 disease progression include plasma viral burden, CD4+ T-cell count, and CD38 expression on CD8 T cells. To better understand whether the predictive value of these markers is dependent on how long an individual has been infected, we analyzed data from the Multicenter AIDS Cohort Study early (median = 2.8 years) and late (median = 8.7 years) in the course of infection. Overall, we found that HIV RNA and CD38 levels were similarly predictive of AIDS early on compared with a relatively weaker CD4 cell count signal. Later in the course of infection, CD38 level remained the strongest predictive marker and CD4 cell count registered a marked increase in prognostic power. Among untreated individuals, there was little difference in prognosis (median time to AIDS) associated with given marker values regardless of infection duration. The prognosis given a specific viral load level tended to deteriorate late in the course of infection among those undergoing treatment with monotherapy or combination therapy, however. These data provide a unique historical look at the predictive value and prognostic significance of HIV-1 disease markers at different stages of infection in a large cohort, with direct relevance to current patients who are untreated or for whom treatment is ineffective. PMID- 11917240 TI - Therapeutic effects of nonnucleoside reverse transcriptase inhibitors on the central nervous system in HIV-1-infected patients. AB - Psychomotor slowing predicts the development of HIV-1-associated dementia, AIDS, and death independent of immune status. We retrospectively selected all patients who showed pathologic psychomotor slowing as a sign of central nervous system (CNS) dysfunction before the onset of therapy and who were then treated with nonnucleoside reverse-transcriptase inhibitors-either efavirenz (EFV) (n = 65 patients) or nevirapine (NVP) (n = 39 patients), each given in combination with two nucleoside analogues (NAs). Patients who were treated only with two NAs (n = 66) served as controls. Patients were observed for 6 months. Both EFV and NVP combinations improved CNS function as determined by electrophysiologic motor tests. The therapeutic effects of EFV and NVP did not depend on the type of NA added. Although results did not reach significance, NVP combinations were more effective than EFV combinations or therapy regimens with NAs alone in patients who were naive to all antiretroviral therapy. EFV and NVP combinations were equally effective in patients pretreated with highly active antiretroviral therapy, including protease inhibitors. PMID- 11917241 TI - Higher rate of toxicity with no increased efficacy when hydroxyurea is added to a regimen of stavudine plus didanosine and nevirapine in primary HIV infection. AB - Twenty-four subjects presenting at a single treatment center with primary HIV infection were enrolled in a pilot study aimed to establish the possible role of hydroxyurea in this setting. Study participants were randomly assigned to receive or not to receive hydroxyurea in addition to stavudine (d4T) plus didanosine (ddI) and nevirapine (NVP). Seventy-five percent of patients without hydroxyurea had plasma HIV RNA below 50 copies/mL at 48 weeks by both intention-to-treat (ITT) and on-treatment (OT) analysis in comparison with 50% (ITT) and 67% (OT) of patients with hydroxyurea (p >.1). A median increase of >200 cells/mm3 was observed from baseline to week 48 whether or not hydroxyurea was included in the regimen. Overall, in 12 patients treated with hydroxyurea, 33 adverse events were reported versus 19 reported for 12 patients who did not receive hydroxyurea (p <.05). Our results suggest that that adding hydroxyurea to a regimen of d4T plus ddI and NVP increases toxicity without improving the antiviral effect. PMID- 11917242 TI - High indinavir Cmin is associated with higher toxicity in patients on indinavir ritonavir 800/100 mg twice-daily regimen. AB - We retrospectively evaluated the incidence of side effects and treatment intervention according to indinavir trough concentration in 63 patients taking indinavir-ritonavir 800/100 mg twice daily. Median indinavir trough concentration was 1446 ng/mL at 800/100 mg twice daily associated with 60% of measured toxicity. Among patients with indinavir trough concentration >500 ng/mL, 46 of 49 had a dosage adjustment and 17 have had more than two dosage adjustments. The primary reason for dosage adjustment was toxicity in 69% (43 cases). Renal and cutaneous toxicity were predominant.After dosage adjustment, median indinavir trough concentration was 459 ng/mL, which was associated with 8% of toxicity. Trough concentrations >500 ng/mL were correlated with increased toxicity (p <.05) and more treatment intervention (p =.02). In conclusion, achievement of indinavir trough concentrations <500 ng/mL appears to be safe, and an optimal concentration range for indinavir trough concentration could be 150 to 500 ng/mL for an indinavir-ritonavir twice daily regimen. PMID- 11917243 TI - Trends in diseases reported on U.S. death certificates that mentioned HIV infection, 1987-1999. AB - To examine trends in the proportions of deaths with various diseases among deaths with HIV infection, we analyzed multiple-cause death certificate data for all deaths in the United States from 1987 through 1999. Disease proportions were adjusted to control for demographic changes. Deaths reported with HIV infection increased from 15,331 in 1987 to 47,977 in 1995 and then decreased to 16,061 in 1999. Among these reported deaths, new trends during the period from 1995 through 1999 included decreases in the proportions with cytomegalovirus disease (from 6.8% to 2.8%), wasting/cachexia (9.8% to 6.8%), and dementia/encephalopathy (6.3% to 3.9%) and increases in the proportions with septicemia/septic shock (from 9.2% to 13.4%) and diseases of the liver (4.9% to 11.6%), kidney (6.3% to 9.1%), and heart (4.2% to 6.9%). Continuations of pre-1995 trends included decreases in the proportions with nontuberculous mycobacteriosis (7.1% to 3.1%) and Kaposi sarcoma (5.3% to 2.6%). Advances in antiretroviral therapy probably caused deaths due to HIV infection to decrease after 1995. Consequently, the proportions of deaths with HIV that were caused by other conditions increased. Improved prophylaxis or treatment of some opportunistic infections could also have reduced the proportions of deaths with those diseases, whereas antiviral drug toxicity could have contributed to increases in the proportions with noninfectious organ diseases. PMID- 11917244 TI - Impact of highly active antiretroviral therapy on individual AIDS-defining illness incidence and survival in Australia. AB - OBJECTIVE: To determine the effect of highly active antiretroviral therapy (HAART) on incidence of initial AIDS-defining illnesses (ADIs) and survival after individual ADIs. METHODS: Australian AIDS notification data over the period 1993 to 2000 were examined. Analyses were based on all initial ADIs. To examine the impact of HAART, two periods of AIDS diagnosis were chosen: pre-HAART (1993-1995) and HAART (1996-2000). Comparisons between these two periods included proportion of individual ADIs, median CD4 lymphocyte counts at and survival following AIDS and individual ADIs. Median survival was based on Kaplan-Meier estimates, with examination of factors influencing survival in a Cox proportional hazards model. RESULTS: Over the period 1993 to 2000 in Australia, 5017 initial ADIs were diagnosed among 4351 AIDS cases. At AIDS diagnosis, changes from the pre-HAART (1993-1995) to HAART (1996-2000) periods included an increased proportion of Pneumocystis carinii pneumonia (PCP) (25.9% to 30.4%; p =.001), AIDS dementia complex (5.2% to 6.8%; p = 0.029), non-Hodgkin lymphoma (NHL) (4.4% to 6.3%; p =.005), and tuberculosis (0.5% to 2.7%; p <.0005). Median survival following AIDS increased from 19.6 months for AIDS cases diagnosed in 1993 to 1995 to 39.6 months for AIDS cases diagnosed in 1996 to 2000 (p <.0005). Median survival was stable for NHL (7.5-8.8 months; p =.26), but increased significantly for almost all other ADIs. CONCLUSIONS: An increased proportion of PCP relative to other ADIs suggests an increasing proportion of AIDS patients not receiving specific prophylaxis, presumably because of "late" HIV diagnosis. Survival following almost all ADIs has increased in the era of HAART, although the prognosis after NHL remains extremely poor. PMID- 11917245 TI - Older children and adolescents surviving with vertically acquired HIV infection. AB - This article describes the characteristics of children infected vertically with HIV surviving 10 years or more who were enrolled in the prospective European Collaborative Study. Thirty-four of 187 infected children were identified with a median age of 11.4 years (range, 10.1-15.9 years). Factors examined included clinical status, immunologic and virologic characteristics, type of antiretroviral therapy, and psychosocial characteristics. By 10 years of age, 6 (18%) children had progressed to Class A as determined by the system of the U.S. Centers for Disease Control and Prevention (CDC), 17 (52%) to class B, 7 (21%) to class C, and 3 (9%) had remained asymptomatic. At 73% (904 of 1234) of scheduled clinic visits, these children had no symptoms of HIV disease. Most children were in CDC immune categories 1 (18, 56%) or 2 (11, 34%) at their last visit. Three quarters (24 patients) were on combination therapy with three or more drugs, although 3 children had never received any antiretroviral therapy. Nineteen (56%) children were living with at least 1 parent and the mothers of 13 (38%) children had died. Most (77%) children had been told about their HIV infection. Children infected vertically with HIV who have survived their first 10 years are mainly free of serious symptoms. As they enter adolescence, additional services are needed including support with disclosure to others, therapy, and sexual health. PMID- 11917246 TI - The price of development: HIV infection in a semiurban community of Ghana. AB - OBJECTIVE: To describe the epidemiology of HIV infection in the Manya Krobo District, Ghana, and its potential link to the building of the Akosombo dam. METHODS: A questionnaire and a blood sample were collected among 1228 consecutive pregnant women seen at the prenatal clinics of the two major hospitals of the district. RESULTS: Overall, prevalence of HIV and of serologically confirmed syphilis were 14.9% and 0.7%, respectively. HIV infection was more prevalent among the Krobo ethnic group (137 of 742 [18.5%]) than among other ethnic groups (46 of 486 [9.5%]; p <.001). Two distinct patterns of HIV distribution were identified. Among the Krobos, HIV was common among all age groups, reached a plateau (21.9%) in the 30- to 34-year-old group and was associated strongly with having lived in Cote d'Ivoire and with having received only primary school education. Among the other ethnic groups, prevalence decreased with age, from 17.2% in the 13- to 19-year-old age group to 1.4% among women aged 35 years or older, and HIV infection was associated with having had first sexual intercourse before the age of 17 years. In logistic regression analysis, the independent risk factors for HIV infection were age, schooling, age at first sexual intercourse; having lived in Cote d'Ivoire; age and schooling showed significant interactions with ethnic group. CONCLUSIONS: The high HIV prevalence documented in this part of Ghana seems to be, to some extent, a consequence of construction of the Akosombo dam in the 1960s. The flooding of the land, the failures of the resettlement program and the ensuing poverty prompted economically driven migration, specially to Cote d'Ivoire, where many migrants became infected with HIV. Local transmission followed. This illustrates that HIV can disseminate widely in a society where most men are circumcised and where genital ulcerative diseases are uncommon and should be an indication for less complacency about HIV control in West Africa. PMID- 11917247 TI - HIV-1 epidemic among female bar and hotel workers in northern Tanzania: risk factors and opportunities for prevention. AB - We conducted this study to determine the prevalence and risk factors for HIV-1 infection among women (N = 312) who were working in the bars and hotels in Moshi, a town in northern Tanzania. Study subjects were interviewed to obtain information about HIV-1 risk factors and examined to collect samples for the diagnosis of sexually transmitted diseases (STDs). The prevalence of HIV-1 was 26.3% (95% confidence interval [CI], 21.4%-31.2%). In multivariate analyses, the risk of HIV-1 increased with increasing age (p value, test for linear trend <.001) and the number of sexual partners during the last 5 years (p value, test for linear trend <.03). Other significant predictors were having a male partner with other sexual partners (Adjusted odds ratio [AOR], 1.92; 95% CI, 1.03-3.60), and consuming alcohol >2 days per week (AOR, 2.56; 95% CI, 1.12-5.88). The risk of HIV-1 was also significantly increased in women with bacterial vaginosis (AOR, 2.37; 95% CI, 1.09-5.13) and in study subjects with herpes simplex virus (HSV)-2 antibodies (AOR, 2.48; 95% CI, 1.24-4.98). These results indicate that women working in these settings were at increased risk of HIV-1. Programs aiming at promoting safer sexual practices and control of other STDs are urgently needed in this population. Such programs should address the underlying conditions that facilitate risk behaviors and create obstacles for these women who wish to protect themselves against HIV-1. PMID- 11917248 TI - Pleural and peritoneal lymphoma among people with AIDS in the United States. AB - OBJECTIVE: To describe the occurrence and characteristics of pleural and peritoneal lymphoma in a large cohort of persons with AIDS in 11 regions in the United States. METHODS: We used AIDS and cancer registries to identify cases of non-Hodgkin lymphoma (NHL) among 304,439 adults with AIDS. NHLs were categorized by site codes into pleural/peritoneal lymphoma and other NHLs. Data on age, sex, HIV exposure category, histology, history of Kaposi sarcoma (KS), CD4 counts, and survival were analyzed. RESULTS: Fourteen lymphomas were identified (four within the pleura, 10 in the peritoneum) representing 0.13% (95% confidence interval [CI], 0.05-0.20) of 10,510 cases of NHL. Those with pleural/peritoneal lymphoma were similar to those with other NHLs in age (median, 43 years), race (79% white, 7% black, 14% Hispanic), and HIV transmission category (86% homosexual men), but they tended to have a higher prevalence of prior KS (29% vs. 12%; p =.06). More cases of pleural/peritoneal lymphoma had immunoblastic histology than did other NHLs (43% vs. 22%; p =.06). CD4 counts for pleural/peritoneal lymphomas were also higher than for other NHLs (median 203 vs. 65 cells/mm3; p =.05), but post-NHL survival was similar (median 7.1 vs. 5.1 months, respectively; p =.32). CONCLUSIONS: Pleural and peritoneal lymphomas are a rare subtype of AIDS associated NHL, occurring with less severe immune deficiency than for other NHLs. The increased frequency among persons with prior KS suggests a common etiology, presumably infection with KS-associated herpesvirus, as found in primary effusion lymphoma. PMID- 11917249 TI - Two HIV-1 epidemics in Argentina: different genetic subtypes associated with different risk groups. AB - This study determined the risk behaviors and viral subtypes of HIV-1 found in 134 heterosexual HIV-seroprevalent maternity patients, 41 of their sexual partners (men who have sex with women [MSW]), and 95 homosexual men (men who have sex with men [MSM]) from Buenos Aires, Argentina. Peripheral blood mononuclear cells (PBMCs) were purified from blood and used for DNA extraction, amplification, and genotyping by the envelope heteroduplex mobility assay (env HMA). Most of the women had been infected by having sex with an already infected partner (84%), whereas most of the male partners had been infected via drug use (76%). Both the patients and their sexual partners were poorly educated, only 30% having completed secondary school. The MSM study subjects, however, were significantly better educated and had a lower prevalence of injecting drug use.Env HMA subtype F was found in 77% (103 of 134) of the maternity patients, with similar rates in their partners (73%). Most of the remaining samples were env subtype B. All but one of the couples was concordant in subtype. In the MSM risk group, however, only 10% were env HMA subtype F. Ninety percent of the MSM samples were subtype B. There are at least two independent epidemics of HIV-1 infection in Buenos Aires, Argentina. One, in heterosexual men and women, is dominated by env subtype F whereas the other, in homosexual men, is dominated by env subtype B, as determined by env HMA. PMID- 11917250 TI - Thymic size and muscle mass of HIV-infected asymptomatic children from Burkina Faso. PMID- 11917251 TI - Treatment of human skin with an extract of Fucus vesiculosus changes its thickness and mechanical properties. AB - Recently the researchers found that an extract of Fucus vesiculosus, which is a type of seaweed, promotes the contraction of fibroblast-populated collagen gels through increased expression of integrin molecules. In this study, they investigated the effects of topical application of an aqueous extract of this alga on the thickness and the mechanical properties of human skin. A gel formulation that included 1% of the extract was applied topically to human cheek skin twice daily for five weeks. A significant decrease in skin thickness measured by B-mode ultrasound was elicited, as was a significant improvement in elasticity measured with a Cutometer as compared with controls. In cheek skin, the thickness normally increases and the elasticity usually decreases with age. These results suggest that the Fucus vesiculosus extract possesses anti-aging activities and should be useful for a variety of cosmetics. PMID- 11917252 TI - Prediction of sun protection factors by calculation of transmissions with a calibrated step film model. AB - Measurements of in vitro sun protection factors (SPFs) are a common way of assessing sunscreen formulations at the stage of screening. The aim of the present investigation is to provide an alternative tool for the estimation of SPF values using a calculation based on the UV spectroscopic properties of the individual UV absorbers. As with in vitro measurements, the crucial step is to work out realistic values of transmissions of UV light through a film of the sunscreen formulation in the important spectral range between 290 and 400 nm. Once these transmissions are given, the SPF can be calculated. Since the human skin is an inhomogeneous substrate, a step film model for the calculation of such transmissions had been proposed by J.J. O'Neill. The step film geometry in this model is a function of two parameters that characterize the fraction of the thin and thick parts of the film and their difference in thickness. The transmissions and therefore the SPF are sensitive functions of the step film parameters. In order to use the model for the prediction of realistic SPF values, the step film parameters are calibrated using three sunscreen standard formulations with well known in vivo SPF. A satisfactory correlation of in vivo SPF values and SPF values calculated with the calibrated step film model using an additional 36 different sunscreen formulations (in vivo SPF values between 3 and 36) is demonstrated. PMID- 11917253 TI - The extract of the flowers of Prunus persica, a new cosmetic ingredient, protects against solar ultraviolet-induced skin damage in vivo. AB - The flowers of Prunus persica Batsch have been used for skin disorders in East Asia from ancient times. In this investigation, the ethanol extract from this plant material was prepared and several major constituents were isolated. In addition, the protective effects of the extract were evaluated against solar ultraviolet (UV)-induced skin damage using in vivo animal models of UVB-induced erythema in guinea pigs and ear edema in ICR mice. From the extract, four kaempferol glycoside derivatives were successfully isolated and their contents were measured with HPLC. Among the derivatives isolated, the content of multiflorin B was highest (3.3%, w/w). The P. persica extract clearly inhibited UVB-induced erythema formation dose dependently when topically applied (IC(50) = 0.5 mg/cm(2)). It also inhibited UVB-induced ear edema (49% inhibition at 3.0 mg/ear). Moreover, multiflorin B inhibited UVB-induced erythema formation (80% inhibition at 0.3 mg/cm(2)), indicating that this compound is one of the active principles of the extract. All these results suggest that P. persica extract may be useful for protection against UVB-induced skin damage when topically applied. PMID- 11917254 TI - Influence of vehicles on the phase transitions of model sebum. AB - It is hypothesized that vehicles that are miscible with sebum may concentrate drugs in the sebaceous follicle. This is important for the treatment of diseases like alopecia and acne. The main objective of the study was to identify different vehicles that affect the thermal behavior of sebum using differential scanning calorimetry (DSC). For this purpose, a model sebum mixture was prepared based on a composition reported in the literature. The test vehicle was added in a concentration of 15% of the weight of the sebum. Small portions of the above mixture were put in a pre-weighed DSC pan. These were run from -50 to 100 degrees C at 5 degrees C/minute. In the model sebum, four distinct transitions were observed: Mp-1 and Mp-2 occurred below 0 degrees C while Mp-3 and Mp-4 occurred above 30 degrees C. Vehicles that affected Mp-3 and Mp-4 were considered for further analysis. From the vehicles tested it was found that the hydrophobic materials were more effective in lowering Mp-3, while the ones that affected Mp-4 did not show any particular trend. Some of the vehicles tested are known skin permeation enhancers, and it is proposed that they interact with sebum and increase permeation by the follicular route. It was found that DSC may be used to identify vehicles that are miscible with sebum and that may deliver drugs preferentially to the sebaceous follicle. PMID- 11917255 TI - Validation of an analytical procedure for the determination of oxidative hair dyes in cosmetic formulations. AB - A high range and variety of cosmetic formulations that contain oxidative hair dyes and matrix-forming compounds have been industrially developed over recent years and are now available on the international market. Member states of the European Union are responsible for conducting analyses of cosmetic products as deemed necessary by law and European regulation enforcement. Therefore, inspection authorities as well as the cosmetics trade and industry need validated analytical methods for the identification, characterization, and/or quality control of specific active ingredients or formulations with the aim of implementing the European Union Cosmetic Directives (76/768/ECC, 95/17/EC). In this frame, we validated a candidate reference method that enables the identification and quantification of hair dye-forming compounds. This method consists of a separation by RP-HPLC coupled with a DAD after a liquid-liquid extraction procedure for separating matrix components from the dye-forming compounds. The validation of the method includes common criteria such as the repeatability of the analysis and the establishment of figures of merit, as well as statistical evaluations and quality assurance in order to follow the recommendations of the Eurachem guide for analytical measurements. PMID- 11917257 TI - Sleep disordered breathing and COPD. AB - This issue of Sleep and Breathing presents a section on sleep in COPD, a widespread disease consuming many health resources which is often diagnosed so late that little chance of reversibility remains. The early detection of the warning clinical signs can include sleep studies, mainly in the presence of arterial carbon dioxide levels higher than expected from pulmonary function tests. Two of the articles deal with hypercapnia and nocturnal hypoventilation in COPD; the third underlines the impact of tobacco smoking on snoring and on oxygen availability to tissues, showing a poor reliability of pulse oximetry in subjects with heavy smoking habits. PMID- 11917258 TI - Long-term effects of treatment with nasal continuous positive airway pressure on lung function in patients with overlap syndrome. AB - We assessed the effects of chronic nasal continuous positive airway pressure (CPAP) therapy on lung function in a series of unselected patients with overlap syndrome, and we determined whether there were differences in the response induced by CPAP between hypercapnic (PaCO2 > or =45 mm Hg) and eucapnic patients with overlap syndrome. The study population included 55 unselected patients (48 men, mean age of 58.5 +/- 10.5 years) with a concurrent diagnosis of chronic obstructive pulmonary disease (COPD) and obstructive sleep apnea-hypopnea syndrome (OSAHS) who had been referred to the Department of Pulmonology of our hospital over 2 consecutive years and in whom work-up studies resulted in the prescription of nasal CPAP therapy. An apnea-hypopnea index (AHI) greater than or equal to 10 in the cardiorespiratory polygraphy was required for the diagnosis of OSAHS. A forced expiratory volume in one second (FEV1) less than 80% and FEV1 forced vital capacity (FVC) ratio less than 70% of the reference values were required for the diagnosis of COPD. Control lung function studies and arterial blood gas measurements were performed at 6 and 18 months of CPAP therapy. These patients with overlap syndrome accounted for 28.5% of all patients with OSAHS treated with CPAP during the study period. The mean AHI was 37.3 +/- 26.1 and the mean CPAP level 7.3 +/- 1.3 cm H2O. Thirty-three patients were hypercapnic (PaCO2 > or = 45 mm Hg) and 22 eucapnic. The hypercapnic group had higher AHI value (44.3 +/- 26.9) than the eucapnic group (28.6 +/- 21.9) (P < 0.05). After 6 months of CPAP therapy, there were statistically significant increases in PaO2, FEV1, and FVC, accompanied by significant decreases in PaCO2, serum bicarbonate levels, and alveolar-arterial oxygen difference. Response of overlap syndrome patients to CPAP therapy was superior in the hypercapnic group, particularly in relation to improvement of arterial blood gases. However, statistically significant differences in all parameters for the comparison between 6 and 18 months were not recorded. PMID- 11917259 TI - Hypercapnia in overlap syndrome: possible determinant factors. AB - We retrospectively evaluated data from 213 consecutive patients; 152 were affected by obstructive sleep apnea (OSA), 29 had OSA associated with chronic obstructive pulmonary disease (COPD), also known as overlap syndrome, and 32 had COPD. Patients with obesity-hypoventilation syndrome were not included. The aims of the study were to evaluate the anthropometric, pulmonary, and polysomnographic characteristics of patients affected by overlap syndrome compared to "simple" OSA and to COPD subjects and to analyze the determinants of hypercapnia in overlap syndrome. In the comparison between overlap and OSA patients, the overlap group had a significantly higher PaCO2 (44.59 vs. 39.22 mm Hg; p < 0.01), in the presence of a similar AHI (40.46 vs. 41.59/h). Comparing overlap to COPD patients, overlap showed a significantly higher PaCO2 value (44.59 vs. 39.63 mm Hg; p < 0.005) and had significantly less severe obstructive impairment (FEV 162.93 vs. 47.31%; FEV1/FVC ratio 66.71 vs. 59.25%; p < 0.005). Anthropometric, pulmonary function, and polysomnographic data did not differ between normo- and hypercapnic overlap patients. The best model (stepwise multiple regression analysis) for predicting PaCO2 in overlap patients showed r2 value 0.65: PaO2 contributed to 38%, FEV1 to 15%, and weight to 12%. In conclusion, the occurrence of hypercapnia in overlap patients is only partially explained by the combination of overweight and reduced respiratory function, supporting the hypothesis of a multifactorial genesis. PMID- 11917260 TI - Cigarette smoking behavior and respiratory alterations during sleep in a healthy population. AB - BACKGROUND AND STUDY OBJECTIVE: The importance of tobacco smoking in the origin of sleep ventilatory abnormalities is disputed. The purpose of our study was to evaluate the influence of cigarette smoking behavior on the sleep respiratory alterations in a healthy population. DESIGN AND METHODS: We studied 38 healthy volunteers (21 M, 17 F; age 42 +/- 12 years; BMI 23.7 +/- 3.6 kg/m(2)) who were divided into two matched groups: current tobacco smokers (n = 18; over 10 pack years) and nonsmokers (n = 20). All individuals underwent a single nocturnal domiciliary polygraphic study (Polygraphics CNS, Minneapolis, Minnesota). Apnea (AI), apnea-hypopnea (AHI), and desaturation (DI) indexes were defined according to conventional criteria. A nocturnal hypoxia index (NHI) was calculated as an index of the magnitude and duration of oxyhemoglobin desaturation during sleep. The mean transcutaneous oxygen saturation (SpO(2)) of the first 60 seconds of oxymetric registration (subject supine and awake) was considered basal SpO(2). Venous carboxyhemoglobin (COHb) levels were measured (CO-Oximeter AVL-912, Basel, Switzerland) in all individuals before (22:00 h) and after (10:00 h) sleep. A correction factor of 0.9 x COHb was applied to the basal SpO(2) values to calculate the corrected basal SpO(2) (SpO(2 corr)). RESULTS: AI, AHI, and DI were not significantly different between smokers and nonsmokers. The smokers have significantly higher NHI than nonsmokers [median (25th percentile-75th percentile): 5.3 (0-39.7) vs. 0.5 (0-1.7); p = 0.017]. There were significant correlations (P < 0.05) in smokers between NHI and pack-years index, between NHI and COHb levels, and between current smoking intensity and COHb levels. As expected, smokers had higher COHb levels at 10:00 as well as at 22:00 hours. The SpO(2 corr) was significantly lower (p < 0.001) among smokers than nonsmokers (88.9 +/- 3.3% vs. 94.7 +/- 1.3%). In multiple regression analyses, AHI and DI showed a significant correlation (p = 0.02 and p = 0.05, respectively) with habitual snoring, and NHI with pack-years and BMI (p = 0.02 and p = 0.04, respectively). CONCLUSIONS: Cigarette smoking does not seem to be associated with increased apneic activity during sleep. However, it is associated with a decrease in nocturnal oxygen saturation. PMID- 11917261 TI - Nocturnal desaturations in COPD: still an open question? AB - The relevance of sleep oxyhemoglobin desaturations in COPD patients is still a matter of debate. The best definition for clinical outcomes is the occurrence of episodes lasting at least 5 minutes and reaching SaO2 levels less than or equal to 85%. Focusing on longer and milder episodes (at least 30% of the night with SaO2 < 90%) did not provide clinical relevance. One possible consequence of desaturation episodes could be a worsening of pulmonary hemodynamics, but O2 dips could just be a marker of a mechanical derangement associated with a worse prognosis. Data confirming the association of these episodes with the clinical type of COPD and with inflammatory mediator modulation are still lacking. PMID- 11917262 TI - On "Snoring in the Ancient World". PMID- 11917263 TI - Snoring in the ancient world. AB - Published in Sudhoffs Archiv fur Geschichte der Medizin und Naturwissenschaften, 1941. PMID- 11917264 TI - Sleep medicine before and after Dickens. AB - Sleep disordered breathing has come to public awareness in the last 10 to 15 years. However, the roots of medical knowledge about sleep disordered breathing reach back several hundred years farther. Greek and Roman publications reported heavy snoring and breathing pauses as diseases that impaired social life. In our day, popular comic strip figures like "Obelix" remind us that the mysterious noise of heavy snoring has been nerve-racking to people since ancient times. PMID- 11917265 TI - Association of sleep and academic performance. AB - Poor school performance by adolescent students has been attributed in part to insufficient sleep. It is recognized that a number of factors lead to diminished total sleep time and chief among these are early school start times and sleep phase delay in adolescence. Political initiatives are gaining momentum across the United States to require later school start times with the intent of increasing total sleep time and consequently improving school performance. Later school start times come with significant costs and impact other activities of families and communities. The decision to implement later school start times cannot be made lightly and deserves support of well-performed research on the impact of these changes. A study evaluating the association of academic performance and total sleep time was performed in middle school and high school students in a suburban Maryland school system. Preliminary results of this study show no correlation of total sleep time with academic performance. Before mandating costly changes in school schedules, it would be useful to perform further research to determine the effects of increasing sleep time on the behaviors of adolescent students. PMID- 11917267 TI - Technical considerations: CT and MR imaging in the postoperative orthopedic patient. AB - Cross-sectional imaging utilizing computed tomography (CT) and magnetic resonance (MR) imaging have become routine components in the imaging assessment of patients with musculoskeletal disease. Unfortunately, in the setting of a postoperative orthopedic patient with associated orthopedic metallic instrumentation, these imaging techniques are prone to artifacts resulting in image quality degradation. An understanding of the physical basis of such metal-related artifacts, and their appearance on CT and MR imaging, has led investigators to the implementation of a series of techniques and modifications to imaging protocols to decrease CT and MR imaging artifacts in the vicinity of metallic instrumentation. Utilizing such modified imaging techniques, consistent, improved CT and MR image quality may be achieved in imaging of the postoperative orthopedic patient. PMID- 11917268 TI - MR imaging of postoperative ligaments of the knee. AB - This article reviews the magnetic resonance (MR) appearance of the ligaments of the knee following surgical repair or reconstruction. The various operative techniques currently used to repair the anterior cruciate ligament, posterior cruciate ligament, and the medial collateral ligament are discussed followed by a description of the normal postoperative MR appearance of both the repair and donor sites. Finally, the MR appearance of various complications at both repair and donor sites is described. PMID- 11917269 TI - MR imaging of the postoperative meniscus of the knee. AB - Surgery for tears of the menisci of the knee has evolved greatly over the last century and is now an extremely common procedure. Meniscal repair and conservative therapy are the preferred methods of treatment, but most symptomatic torn menisci do not meet the requirements for these options and necessitate partial meniscectomy. After surgery, patients often have recurrent pain and need additional diagnosis. Conventional magnetic resonance imaging (MRI) in the postoperative patient is very helpful at diagnosing extrameniscal pathology. It has lower sensitivity for detecting retears after partial meniscectomy, however, as it relies primarily on demonstration of fluid entering the meniscal retear on T2-weighted images. Similarly, conventional MRI is less accurate after meniscal repair, in which the repair site usually maintains altered signal for years. MR arthrography has higher sensitivity for detecting retears in menisci and is the procedure of choice in many situations. PMID- 11917270 TI - Spiral CT arthrography of the postoperative knee. AB - Computed tomography (CT) has gained multiplanar capability and submillimeter spatial resolution due to the development of spiral acquisition mode and multidetector row technology. Multidetector spiral CT arthrography of the knee is a valuable imaging modality for the assessment of lesions of the meniscus, anterior cruciate ligament (ACL), and articular cartilage. This article presents the value of spiral CT arthrography in the assessment of the postoperative knee with emphasis on the postoperative meniscus, articular cartilage, and ACL graft. This technique may be proposed as an alternative to magnetic resonance arthrography. PMID- 11917271 TI - MR imaging after surgery for musculoskeletal neoplasm. AB - Magnetic resonance (MR) imaging is the current radiologic method of choice for both detecting recurrent musculoskeletal neoplasm after surgical resection and defining its anatomic extent within soft tissue and bone. Various factors (such as the presence of surgical hardware and postsurgical seromas, hematomas, edema, scarring, and anatomic distortion) complicate the interpretation of postoperative MR imaging in these patients. By optimizing the MR imaging protocol, integrating relevant clinical and pathologic information (such as the date and extent of the most recent surgery and the histologic type and grade of the original tumor) during interpretation of the images, and being familiar with the typical manifestations of postsurgical changes and recurrent musculoskeletal tumors, the radiologist can maximize his ability to help guide patient management effectively. PMID- 11917272 TI - Musculoskeletal sonography of the postoperative orthopedic patient. AB - Musculoskeletal sonography has an important role in the evaluation of the postoperative orthopedic patient. One major advantage of sonography over magnetic resonance imaging and computed tomography is that artifact from metal, which is common after orthopedic surgery, is relatively limited. This allows a clear assessment of the soft tissues for abnormalities such as tendon tear, tenosynovitis, joint effusion, and soft tissue infection. Sonography also has a role in evaluation of recurrent soft tissue tumor, bone healing, and limb amputation. This article first discusses the basic technical aspects of musculoskeletal sonography and is followed by sonographic examples of normal anatomy. Common applications of musculoskeletal sonography in evaluation of the postoperative orthopedic patient are reviewed, including sonography after tendon surgery, assessment of soft tissues adjacent to orthopedic hardware, recurrent musculoskeletal soft tissue tumors, sonography of bone healing, and evaluation of limb amputation. Guidelines and pitfalls of these applications are reviewed. PMID- 11917273 TI - MR imaging of joint arthroplasty. AB - Imaging evaluation of the painful total joint arthroplasty has, in the past, been limited to conventional radiographs, arthrography, and nuclear scintigraphy. All three modalities provide some clinical information regarding the status of the prosthesis, adjacent bone, and surrounding soft tissues but are hampered by poor specificity, particularly nuclear scintigraphy. Magnetic resonance imaging, using optimized pulse sequences to reduce the susceptibility artifact generated by the arthroplasty, can provide clinically relevant information regarding the prosthesis as well as the surrounding bone, soft tissues, and neurovascular structures. PMID- 11917275 TI - Proceedings from the Kimmel Cancer Center Symposium, The 1st Investigators' Congress on Radioprotection. June 7-10, 2001, St Croix, US Virgin Islands. PMID- 11917276 TI - The evolution and application of toxicity criteria. AB - The assessment and reporting of toxicity plays a central role in oncology. The foundation of toxicity reporting is the toxicity criteria system. Multiple systems have been developed and have evolved substantially since first introduced more than 20 years ago. The wide adoption of a standardized criteria system can facilitate comparison between institutions and clinical trials. The Common Toxicity Criteria (CTC) system was revised and expanded substantially in 1997 and now includes acute radiation criteria as part of multimodality grading. In contrast to the progress made in standardizing acute effects, the current use of multiple late effects grading systems by different groups hinders comparability of clinical trials, impedes the development of toxicity interventions, and encumbers the proper recognition and reporting of late effects. Just as the revised CTC has provided a new common standard for the grading of acute effects, a similar effort is needed in the late effects area. The creation of a unified late effects system is under active development. PMID- 11917274 TI - A missense mutation in the LIM2 gene is associated with autosomal recessive presenile cataract in an inbred Iraqi Jewish family. AB - In an inbred Iraqi Jewish family, we have studied three siblings with presenile cataract first noticed between the ages of 20 and 51 years and segregating in an autosomal recessive mode. Using microsatellite repeat markers in close proximity to 25 genes and loci previously associated with congenital cataracts in humans and mice, we identified five markers on chromosome 19q that cosegregated with the disease. Sequencing of LIM2, one of two candidate genes in this region, revealed a homozygous T-->G change resulting in a phenylalanine-to-valine substitution at position 105 of the protein. To our knowledge, this constitutes the first report, in humans, of cataract formation associated with a mutation in LIM2. Studies of late-onset single-gene cataracts may provide insight into the pathogenesis of the more common age-related cataracts. PMID- 11917277 TI - Amifostine in simultaneous radiochemotherapy of advanced head and neck cancer. AB - The authors discuss the results of 3 studies of their group reflecting the possible role of amifostine in simultaneous radiochemotherapy (RCT) of advanced head and neck cancer. In a controlled phase II trial (1995 through 1996), 39 patients were included in this pilot investigation. A control group (n = 14) received simultaneous RCT of the head and neck region with an irradiation dose of 60 Gy and 2 cycles of carboplatin (700 mg/m(2) cumulative dose). Twenty-five patients received the same basic therapy and an additional 500-mg dose of amifostine before each chemotherapy. Amifostine was administered less than 45 minutes before the end of radiotherapy. The authors observed a dramatic reduction of typical radiotherapy-associated toxicities (mucositis, xerostomia, loss of taste, dysphagia). The hematologic side effects (leukocytopenia, anemia, thrombocytopenia) also were decreased significantly. The overall survival rate and locoregional control of both groups were comparable after 12 months. In a controlled intensification trial (1997 through 1999), the authors included 76 consecutive patients (69 men, 7 women) with pharyngeal cancer (oropharynx, n = 33; hypopharynx, n = 43). The tumors were characterized as unresectable and locally advanced without distant metastasis. All patients received a conventional radiotherapy (2-Gy single dose, daily fractionation) up to doses of 60 Gy and an additional 10 Gy as a boost in the tumor-infiltrated region. A dose of carboplatin, 70 mg/m(2), was given to a group of 45 patients on days 1 through 5 and 29 through 33 of radiotherapy (RCT arm). The resulting cumulative dose was 700 mg/m(2). A group of 31 patients (RCTintens arm) received the same dose of carboplatin on days 1 through 5, 22 through 26, and 43 through 47 or 1 through 5, 15 through 19, 29 through 33, and 43 through 47 of radiotherapy (cumulative dose 1.05 to 1.40 mg/m(2)). All patients received 500 mg of amifostine before each carboplatin administration. If the tumor volume was less than 20 cm(3), we observed an increased 1-year overall survival rate (91% v 71%) and time to progression (17 months v 10 months). If the tumor volume was greater than 20 cm(3), we observed comparable treatment results in both groups (1-year survival rate, 60% v 61%; time to progression, 13 months v 12 months). In a long-term follow-up investigation (1999 through 2000), 531 patients (89 women, 442 men) were analyzed according to their toxicities during regular follow-up investigations at our outpatient facility. All patients were treated by surgery or radio(chemo)therapy because of an advanced head and neck cancer. A total of 218 of 531 patients received the antineoplastic therapy without cytoprotection. An additional 313 patients received their RCT combined with amifostine administration before administration of the radiosensitizer. A significant influence of cytoprotection was registered in the following toxicities: xerostomia, fibrosis, loss of taste, and dysphagia. No impact was seen on the development of interstitial lymph edema and esophageal stenosis. Amifostine could be integrated in simultaneous radiochemotherapy of advanced head and neck cancer patients. The authors favor the administration of amifostine before chemotherapeutics alone. Selective cytoprotection could decrease the main acute toxicities (mucositis, xerostomia, dysphagia) as well as late side effects (xerostomia, loss of taste, fibrosis) of this form of combined treatment. The enhanced therapeutic index may be changed into a prognostic benefit for selected patients with unresectable tumors, if the volume is smaller than 20 cm(3). PMID- 11917278 TI - A prospective, nonrandomized study of the impact of amifostine on subsequent hypothyroidism in irradiated patients with head and neck cancers. AB - The daily administration of subcutaneous amifostine to patients with head and neck cancers before each radiation fraction may reduce the long-term incidence of hypothyroidism (HT) after radiotherapy to the neck and may provide an additional indication for amifostine (Ethyol) use. Cancers of the head and neck afflict 40,000 patients yearly in the United States, and radiotherapy plays a pivotal role in the management of at least half of these patients. Patients commonly are treated with radiation alone, combined surgery and radiation, or definitive chemoradiotherapy. Local control of disease is achieved in many patients who are at risk for late sequelae of treatment that may diminish quality of life. PMID- 11917279 TI - A phase II trial of subcutaneous amifostine and radiation therapy in patients with head and neck cancer. AB - This phase II trial was designed to verify that subcutaneous (SC) administration of Amifostine (Ethyol) protects against radiation therapy (RT)-induced xerostomia and ameliorates amifostine-related side effects (including nausea, vomiting, and hypotension). Patients receiving amifostine SC plus RT had a 56% incidence of acute xerostomia, comparable with previous phase III data with intravenous administration of amifostine. There was good tolerability, with cutaneous toxicity as the most significant side effect. These data suggest that amifostine SC provided comparable protection against RT-induced acute xerostomia as amifostine intravenously. PMID- 11917280 TI - Protection of salivary function by intensity-modulated radiation therapy in patients with head and neck cancer. AB - The degree of xerostomia has been reported to depend on the radiation dose and the salivary gland volume irradiated. Sparing salivary function can be achieved by reducing radiation dose to the salivary glands or using a radiation protector, such as amifostine (Ethyol). In this report, the author reviews clinical experiences in intensity-modulated radiation therapy (IMRT) for head and neck cancer. In experiences, the dosimetric advantage of IMRT did translate into significant reduction of late salivary toxicity in patients with oropharyngeal carcinoma. The author has found no adverse impact on tumor control and disease free survival in patients treated with IMRT. Further, when studying the dose response of parotid gland after irradiation, it was found that the stimulated saliva flow 6 months after IMRT treatment reduced at approximately 4% per Gy exponentially of the mean parotid dose. The authors also review existing clinical data on the combination of amifostine and radiation and the potential therapeutic gain in combining IMRT with amifostine. PMID- 11917281 TI - Radiation pneumonitis and early circulatory cytokine markers. AB - Radiation pneumonitis is a distinct clinical entity that differs from other pulmonary symptoms such as allergic pneumonitis, chemical pneumonitis, or pneumonia by various infectious agents. Recent research has supported the mechanism of cellular interaction between lung parenchymal cells and circulating immune cells mediated through a variety of cytokines including proinflammatory cytokines, chemokines, adhesion molecules, and profibrotic cytokines. Identifying reliable biomarkers for radiation pneumonitis will allow us to identify individuals at risk for pneumonitis before or during the early stage of therapy. Prospective blood sampling, scoring of respiratory symptoms, and chest imaging were conducted for patients receiving thoracic radiotherapy for malignancy. Serial plasma specimens were analyzed for circulating cytokine changes before, during, and up to 12 weeks after radiation. Radiation pneumonitis was diagnosed using National Cancer Institute (NCI) common toxicity criteria. Cytokine analysis was assayed for interleukin 1alpha (IL-1alpha), interleukin 6 (IL-6), monocyte chemotactic protein 1 (MCP-1), E-selectin, L-selectin, transforming growth factor beta1 (TGF-beta1), and basic fibroblast growth factor (bFGF) using enzyme linked immmunosorbant assay (ELISA). Twenty-four patients had clinical follow-up longer than 12 months after radiotherapy. Thirteen had symptomatic pneumonitis (NCI grade 2). The peak incidence of symptoms was between 6 and 13 weeks after radiotherapy. Six patients had only radiographic infiltrates (NCI grade 1). Five patients did not have clinical or radiographic pneumonitis. Both IL-1alpha and IL 6 levels were significantly higher before, during, and after radiotherapy for those who had pneumonitis. The pattern of changes of MCP-1, E-selectin, L selectin, TGF-beta1, and bFGF varied, but none of these cytokines correlated with radiation pneumonitis. Analysis of a panel of circulating cytokines with different putative functions in radiation pulmonary injury identified IL-1alpha and IL-6 as early circulating cytokine markers for radiation pneumonitis. PMID- 11917282 TI - Phase II: trial of twice weekly amifostine in patients with non-small cell lung cancer treated with chemoradiotherapy. AB - Twenty-four patients with non-small cell lung cancer received induction chemotherapy (paclitaxel, carboplatin) followed by concurrent thoracic irradiation (RT) and weekly paclitaxel. Acute esophagitis was scored weekly. Amifostine (AMI), 500 mg intravenously twice weekly, was added to the regimen in the second cohort of 12 patients. AMI was well tolerated. The incidence of grade 3 esophagitis was 18% in the initial 11 patients versus 9% in the AMI-treated patients. Mean esophagitis index (EI) was numerically lower in the AMI-treated patients than in the initial group (5.1 v 11.6, P =.14). The length of esophagus in the RT field was similar in both groups. Median survival time for all patients was 12.4 months. The EI, a novel measure of the severity and duration of acute esophagitis, may be reduced in lung cancer patients receiving AMI twice weekly with thoracic RT and paclitaxel. The effect of AMI was not caused by the shorter irradiated esophageal length. A phase III randomized trial is now open to assess the effect of AMI on esophagitis. PMID- 11917283 TI - Exploring the role of the radioprotector amifostine in locally advanced non-small cell lung cancer: Radiation Therapy Oncology Group trial 98-01. AB - The ultimate goal of any strategy in oncology is to enhance the therapeutic ratio, defined as tumor cell kill divided by normal tissue injury. Whereas most trials focus on intensifying therapy, this often increases toxicity so that there is no overall gain in the therapeutic ratio. Radiation Therapy Oncology Group (RTOG) trial 98-01, a phase III study testing the ability of a radioprotector (Amifostine [Ethyol, WR-2721]) to reduce the toxicity of an intensive chemoradiation regimen for locally advanced non-small cell lung cancer (NSCLC), is unique in that it addresses both "sides" of this equation. During the 1990s, thoracic radiotherapy (RT) combined with chemotherapy was accepted as a "new" gold standard for patients with good performance status locally advanced/inoperable NSCLC. This paradigm shift away from RT alone has raised several key questions: What is the optimal method of integrating chemotherapy and RT? Equally importantly, how can the toxicity of combined modality strategies be reduced? This article will review the background underlying RTOG 98-01, a trial exploring the potential role of amifostine in NSCLC. PMID- 11917285 TI - Radiotherapy or chemotherapy followed by radiotherapy with or without amifostine in locally advanced lung cancer. AB - Radiotherapy (RT) or radiochemotherapy is the treatment of choice for patients with medically or technically inoperable non-small cell lung cancer (NSCLC) localized to the primary site and regional lymph nodes. Radiation-induced damage has been recognized as a major complication in thoracic RT. The use of concurrent chemoradiation has been associated with an increase in acute and late toxicity. Amifostine (Ethyol) is an effective cytoprotective agent and also may have a role in protecting healthy lung tissue during radiation treatment. The purpose of these 2 clinical trials was to investigate whether daily pretreatment with amifostine could reduce the incidence of esophagitis, and acute and late lung toxicity without affecting the antitumor efficacy of the treatment. The first was a phase III randomized trial of 146 patients with locally advanced lung cancer. All patients received conventional RT to a total of 55 to 60 Gy, and they were assigned randomly to pretreatment with 340 mg/m(2) of amifostine (A). Acute and late toxicities were graded according to the Radiation Therapy Oncology Group (RTOG) grading system from grades 0 to 4. Ninety-seven patients were evaluated 2 months post-RT for the incidence of pneumonitis; 43% (23 of 53) of patients in the RT arm and 9% (9 of 44) in the A plus RT arm experienced grade > or = 2 pneumonitis (P <.001). Forty-nine percent (26 of 53) of patients in the RT arm and 16% (7/44) in the A plus RT arm showed changes that were representative of grade > or = 2 lung damage in the computed tomography (CT) scan. Fibrosis was present in 53% (19 of 36) of patients receiving RT versus 28% (9 of 32) in the A plus RT arm at 6 months (P < 0.05). The incidence of esophagitis grade > or = 2 during week 4 was 42% (31 of 73) in the RT arm versus 4% (3 of 73) in the A plus RT arm (P <.001). Among 97 patients evaluable for response 2 months after RT, complete or partial responses were present in 76% (40 of 53) of patients in the RT arm and 75% (33 of 44) in the A plus RT arm (P = 1.0). The second trial was a phase II randomized study of 45 patients with NSCLC. All patients had received platinum-based induction chemotherapy before being referred for conventional radiation treatment with or without A; a total dose of 55 to 60 Gy was administered at the primary site. Acute and late toxicities were evaluated and graded according the RTOG criteria from grades 0 to 4. Forty-five patients were evaluable for response 2 months after RT. Complete or partial responses were achieved in 78% (18 of 23) of patients in the RT arm and 82% (18 of 22) in the A plus RT arm (P =.278). By week 5, 74% (17 of 23) of patients in the RT group versus 36% (8 of 22) in the A plus RT group experienced grade > or = 2 esophagitis. (During the follow-up period, pulmonary toxicity was evaluated by CT scan.) Three months after RT, 65% (15/23) of patients in the RT group and 32% (7 of 22) in the A plus RT group presented with grade > or = 2 pneumonitis (P =.038). Amifostine reduces the incidence of acute and late radiation-induced toxicities. PMID- 11917284 TI - Randomized phase III study of chemoradiation with or without amifostine for patients with favorable performance status inoperable stage II-III non-small cell lung cancer: preliminary results. AB - A prospective randomized study was conducted to determine whether amifostine (Ethyol) reduces the rate of severe esophagitis and hematologic and pulmonary toxicity associated with chemoradiation or improves control of non-small cell lung cancer (NSCLC). Sixty patients with inoperable stage II or III NSCLC were treated with concurrent chemoradiotherapy. Both groups received thoracic radiation therapy (TRT) with 1.2 Gy/fraction, 2 fraction per day, 5 days per week for a total dose 69.6 Gy. All patients received oral etoposide (VP-16), 50 mg Bid, 30 minutes before TRT beginning day 1 for 10 days, repeated on day 29, and cisplatin 50 mg/m(2) intravenously on days 1, 8, 29, and 36. Patients in the study group received amifostine, 500 mg intravenously, twice weekly before chemoradiation (arm 1); patients in the control group received chemoradiation without amifostine (arm 2). Patient and tumor characteristics were distributed equally in both groups. Of the 60 patients enrolled, 53 were evaluable (27 in arm 1, 26 in arm 2) with a median follow-up of 6 months. Median survival times were 26 months for arm 1 and 15 months for arm 2, not statistically significantly different. Morphine intake to reduce severe esophagitis was significantly lower in arm 1 (2 of 27, 7.4%) than arm 2 (8 of 26, 31%; P =.03). Acute pneumonitis was significantly lower in arm 1 (1 of 27, 3.7%) than in arm 2 (6 of 26, 23%; P =.037). Hypotension (20 mm Hg decrease from baseline blood pressure) was significantly more frequent in arm 1 (19 of 27, 70%) than arm 2 (1 of 26, 3.8%; P =.0001). Only 1 patient discontinued treatment because of hypotension. These preliminary results showed that amifostine significantly reduced acute severe esophagitis and pneumonitis. Further observation is required to assess long-term efficacy. PMID- 11917286 TI - Phase II evaluation of amifostine as an esophageal mucosal protectant in the treatment of limited-stage small cell lung cancer with chemotherapy and twice daily radiation. AB - For limited-stage small cell lung cancer, twice-daily radiation with concurrent chemotherapy improves survival rate, but has dose-limiting esophageal toxicity. The authors studied 34 patients treated with amifostine in an attempt to decrease the incidence and grade of esophagitis. The results indicate that there was no reduction in toxicity, but the authors were able to maintain the high complete response rate that had been reported previously. These results differ from the use of amifostine in non-small cell lung cancer in which there is the observation of esophageal protection. PMID- 11917287 TI - Esophageal cancer and the esophagus: challenges and potential strategies for selective cytoprotection of the tumor-bearing organ during cancer treatment. AB - Esophageal cancer is treated optimally with a combined-modality approach according to most clinical investigators. Cytotoxic chemotherapy and ionizing radiotherapy, given in a concomitant schedule, has yielded superior survival rates compared with radiotherapy alone. However, mucosal toxicity from such treatment may compromise quality of life and may mandate an unscheduled break in therapy in some patients who do not respond readily to standard treatments such as antacids; combinations of viscous xylocaine, aluminum hydroxide-magnesium carbonate, and diphenhydramine hydrochloride; oral liquid morphine sulfate, hydrocodone bitartrate, or acetaminophen. Hence, a number of alternative strategies that are designed to either prevent or limit toxicity to normal tissues without diminishing the antitumor effect are being tested. These include the use of conformal radiotherapy treatment planning techniques, amifostine (Ethyol, WR-2721), gene therapy via intratumoral injection of manganese superoxide dismutase-plasmid/liposome, glutamine, melatonin, omega-3 polyunsaturated fatty acids, transforming growth factor, flavonoid compounds, probucol, and keratinizing growth factor. An ongoing phase 2 trial by the North Central Cancer Treatment Group (NCCTG) may help clarify a role for cytoprotectants in patients receiving combined-modality therapy for esophageal cancer. PMID- 11917288 TI - The potential role of amifostine in the treatment of carcinoma of the uterine cervix: a review. AB - Carcinoma of the uterine cervix affects approximately 13,000 women annually in the United States. Radiotherapy is a commonly applied therapy, either as definitive treatment or combined with surgery. Although highly effective, radiotherapy can be associated with significant complications. Amifostine has been shown to be an effective radioprotector in head and neck malignancies. Preliminary evidence suggests a role in cervical cancer. PMID- 11917289 TI - Trade-off to low-grade toxicity with conformal radiation therapy for prostate cancer on Radiation Therapy Oncology Group 9406. AB - The aim of this study was to evaluate and compare the rates of grade 2 or worse late effects in patients treated for prostate cancer on Radiation Therapy Oncology Group (RTOG) 9406. The authors previously have reported the results of patients treated on the first 2 dose levels of this study with respect to grade 3 or greater late toxicity. This analysis examines the incidence of grade 2 toxicity in this study. From August 1994 to September 1999, 424 patients were entered on this dose escalation trial of 3-dimensional conformal radiation therapy (3D CRT) for localized adenocarcinoma of the prostate at doses of 68.4 Gy (level I) and 73.8 Gy (level II). All radiation prescriptions were a minimum dose to a planning target volume. Patients were stratified according to clinical stage and risk of seminal vesicle invasion based on Gleason score and presenting prostate-specific antigen. Average time at risk after completion of therapy ranged from 33.1 to 40.1 months for patients treated at dose level I and 15.6 to 34.2 months for patients at dose level II. The frequency of late effects > or = grade 2 was compared with a similar group of patients treated on RTOG studies 7506 and 7706 with adjustments made for the interval from completion of therapy. The RTOG toxicity scoring scales for late effects were used. The rate of grade 3 or greater late toxicity continues to be low compared with RTOG historical controls. No grade 4 or 5 late complications were reported in any of the 406 evaluable patients during the period of observation. Interestingly, the incidence of grade 2 late toxicity was increased relative to historical controls in all groups and dose levels. In group 1, level I and group 3, level II, the increase in grade 2 complications was statistically significant; 16 complications were observed in group 1, level I when 9.2 were expected (P =.026) and 22 were observed in group 3, level II when 7.6 were expected (P <.0001). When examining all late effects > or = grade 2, there were no significant differences in the rate of late effects in both groups and both dose levels with the exception of group 1, level II. This, in combination with the statistically significant decrease in late effects > or = grade 3, suggests that in most circumstances there has been a shift of grade 3 complications to grade 2. In group 1, dose level II there was a statistically significant reduction in > or = grade 2 late effects, suggesting there was no shift from grade 3 to grade 2 in these patients. In this circumstance there may have been a global reduction in all complications or a shift to late effects less severe than grade 2. In group 2, dose level II there is a trend (P =.085) toward this same result. It is important to continue to examine late effects closely in patients treated on RTOG 9406. The primary objective of dose escalation without an increase rate of > or = grade 3 complications has been achieved. However, the reduction in grade 3 complications may have resulted in a higher incidence of grade 2 late effects. Because grade 2 late effects may have a significant impact on a patient's quality of life, it is important to reduce these complications as much as possible. Improved conformal treatment delivery with intensity-modulated radiation therapy or the use of radioprotective agents could be considered. Clinical trials should use quality-of life measures to determine that trade-offs between severity and rates of toxicity are acceptable to patients. PMID- 11917291 TI - Rationale for a phase I/II radiation dose-escalation study with concurrent amifostine (Ethyol) and infusional 5-FU chemotherapy for preoperative treatment of unresectable or locally recurrent rectal carcinoma. AB - Rectal adenocarcinoma almost invariably requires extirpative surgery to be cured. Although radiation therapy and concurrent chemotherapy often will elicit dramatic responses, the treatment morbidity limits the tolerable dose of radiation to about 45 to 50 Gy. Strategies to reduce procto/enteritis may permit higher doses of radiation to be delivered. This report reviews a recently activated radiation dose-escalation study that combines the radioprotector amifostine (Ethyol, WR 2721) with 3-dimensionally planned radiation therapy and concurrent infusional 5 fluorouracil chemotherapy. PMID- 11917290 TI - Intrarectal application of amifostine for the prevention of radiation-induced rectal injury. AB - Clinically symptomatic late injury to the rectal wall occurs in about one third of patients with prostate cancer treated with external beam irradiation. Reducing the physical dose to the anterior rectal wall without a similar reduction in the posterior peripheral zone is difficult because of the proximity of these structures. Based on our previous observations that intrarectal application of amifostine resulted in very high concentrations of amifostine and its active metabolite WR-1065 in the rectal wall of Copenhagen rats, the authors initiated a phase I clinical trial in 1998. Twenty-nine patients with localized prostate cancer were accrued. Eligibility criteria included histologically confirmed adenocarcinoma, a Karnofsky performance status of > or =70, and no pelvic lymphadenopathy or distant metastases. The total dose to the prostate was 70.2 Gy (20 patients) and 73.8 Gy (9 patients). Therapy was delivered using a 4-field axial technique and 3-dimensional conformal planning. Amifostine was administered intrarectally as an aqueous solution 30 minutes before irradiation on the first 15 days of therapy. Amifostine dose was escalated, in cohorts, from 500 mg to 2,500 mg. Toxicity was evaluated using the Radiation Therapy Oncology Group late morbidity scale. All patients completed therapy with no amifostine-related toxicity at any dose level. The application was feasible and well tolerated. With a median follow-up time of 21 months, 9 patients (33%) had rectal bleeding (8 grade 1, 1 grade 2). Four patients (14%) had symptoms suggestive of radiation injury, which proved to be secondary to nonrelated processes. These included preexisting nonspecific proctitis (1 patient), diverticular disease of the sigmoid colon, rectal polyp (1 patient), and ulcerative colitis (1 patient). Symptoms developed significantly more often in patients receiving 500 to 1,000 mg than in patients receiving 1,500 to 2,500 mg amifostine (7 of 14 [50%] versus 2 of 13 [15%]; P =.0325, 1-sided chi(2) test). Intrarectal application of amifostine is feasible and well tolerated. A complete lack of systemic toxicity obviates the need for close monitoring of patients during and after administration. Rectal symptomatology after external beam radiotherapy to the pelvis cannot be assumed to reflect late radiation damage, because it often is a manifestation of an unrelated pathologic process. The preliminary efficacy data are encouraging and suggest that intrarectal administration of amifostine may reduce radiation damage. Further clinical studies are warranted. PMID- 11917292 TI - Statistical issues in studies of toxicity modifiers. AB - The primary focus of oncology is the search for a cure. In pursuit of this goal, therapeutic regimens have increased in intensity, often causing increased normal tissue morbidity. Pharmacologic agents are being developed to ameliorate or prevent these toxicities. The stepwise investigation of these agents requires a different paradigm from standard drug development. Traditionally, phase I studies are designed to find the maximum tolerated dose of an experimental drug; however, in pharmacologic agents that aim to reduce toxicity, defining the maximum tolerated dose, may not be necessary. The focus of phase I studies in these agents is the establishment of the maximum effective dose. Phase II studies traditionally are designed to establish efficacy of an experimental regimen before advancing to a phase III study. To determine potential efficacy, predictable efficacy levels must be available in the literature for standard therapy. In this manner, the phase II study can be evaluated against the historical control to determine if a phase III study should be conducted. This generally is not possible in the context of toxicity modifiers. Because of variability in toxicity assessment, prior rates of toxicity may not be reliable when evaluated against a phase II study focusing on toxicity amelioration. Because the assessment of toxicity is subjective, retrospective comparisons may be biased and lead to the wrong conclusion. One method to correct for this bias is to conduct placebo-controlled, randomized phase II studies. Phase III studies of toxicity modifiers have toxicity amelioration or prevention as the primary objective. These studies should be designed to assess the subjective and objective components of toxicity, evaluate quality of life, and assess possible adverse effects on therapeutic outcomes. Design and analysis issues are discussed in this report. PMID- 11917293 TI - Preclinical modeling of improved amifostine (Ethyol) use in radiation therapy. AB - Amifostine (Ethyol) has been evaluated clinically as a radioprotective agent for the prevention of xerostomia and mucositis for patients receiving radiotherapy (RT). Currently, amifostine is approved for the prevention of xerostomia in head and neck cancer patients receiving RT when administered intravenously (IV) before RT. For the clinician, there would be several advantages to administering the drug subcutaneously and to being able to show its protective effects on mucositis. The authors have developed a rat RT model to examine the protective effects of amifostine after IV and subcutaneous (SC) administration in a mucositis model. Rats (5 per group) were given 200 mg/kg (human dose equivalent of approximately 1,300 mg/m(2)) of amifostine either IV or SC, and their head and neck regions were exposed to 15.3 Gy of gamma radiation 0.5, 2, 4, and 8 hours after amifostine administration. For 10 days after treatment, the oral cavities of the rats were examined for signs of mucositis. Mucosal erythema and mucosal edema were scored according to 0 through 5 and 0 through 2 scales, respectively, with the scores added to indicate overall mucositis. The average mucositis score for the untreated animals was 3.5. Rats were protected from mucositis up to 4 hours when given amifostine either IV or SC. Rats that received amifostine SC, but not IV, were protected from mucositis 8 hours after administration. Preliminary pharmacokinetic data have revealed slightly higher active metabolite (WR-1065) levels in the parotid gland and small intestine in the rats given amifostine SC compared with IV and equivalent levels in the plasma and kidney. The data showed that SC administration of amifostine gave radioprotection comparable to IV administration up to 4 hours before RT and may be more effective than IV administration at longer pretreatment intervals. PMID- 11917294 TI - Differential activation of nuclear transcription factor kappaB, gene expression, and proteins by amifostine's free thiol in human microvascular endothelial and glioma cells. AB - The effects of WR1065 (SH), the free thiol form of amifostine, on nuclear transcription factor kappaB (NFkappaB) activation, manganese superoxide dismutase (MnSOD) gene expression, and secretion of human vascular endothelial cell growth factor (hVEGF), basic fibroblast growth factor (bFGF), tumor necrosis factor alpha (TNF-alpha), vascular cell adhesion molecule-1 (VCAM-1), intercellular adhesion molecule-1 (ICAM-1), E-selectin, P-selectin, and interleukins IL-1alpha, IL-6, and IL-8 were investigated and compared in human microvascular endothelial (HMEC) and human glioma cells. WR1065 was evaluated at 2 concentrations, 4 mmol/L, ie, its most effective cytoprotective dose, and 40 micromol/L, a noncytoprotective but highly effective dose capable of preventing radiation and chemotherapeutic drug-induced mutations in exposed cells. A 30-minute exposure of HMEC and glioma cell lines U87 and U251 to WR1065 at either of the concentrations resulted in a marked activation of NFkappaB as determined by a gel shift assay, with the maximum effect observed between 30 minutes and 1 hour after treatment. Using a supershift assay, WR1065 exposure was observed to affect only the p50-p65 heterodimer, and not the homodimers or heterodimers containing p52 or c-Rel subunits of NFkappaB. WR1065 was also found to enhance MnSOD gene expression in both HMEC and glioma cells. Gene expression was enhanced 1.8-fold over control levels in HMEC over a period ranging from 12 to 24 hours after the time of maximum activation of NFkappaB. In contrast, MnSOD gene expression in U87 cells rose 3.5 times above control levels over this same period. WR1065 had no effect on the levels of adhesion molecules, cytokines, and growth factors secreted by cells exposed for up to 24 hours as measured by enzyme-linked immunosorbent assay. PMID- 11917295 TI - Behavior modification: a nursing approach for young children with atopic eczema. AB - Managing atopic eczema represents one of the most challenging aspects of dermatology nursing, especially in children. A main contributing factor to the difficulty in management is the unrelenting pruritus experienced by sufferers. A practice model for behavior modification can be an effective nursing strategy in the management of pruritus and scratch associated with atopic eczema. PMID- 11917296 TI - Creative stress-management techniques for self-renewal. AB - The daily stress and pressure of today's nurses are profound. As workloads escalate and staffing declines, nurses must be proactive and develop healthier lifestyles. Creative stress management techniques offer health-promoting strategies to manage stress in nurses' lives. PMID- 11917297 TI - What's your assessment? Erythrasma. PMID- 11917298 TI - Leading from the inside out. PMID- 11917299 TI - Nursing in the new millennium: challenges and opportunities. AB - The technological, social, political, and economic pressures in our world are placing inordinate strains on the health care system. Nurses, as the largest groups of workers in health care, are shaped by these changes in the system and also have considerable opportunity to shape the system itself. Meeting the challenges of the 21st century will require a significant dedication to learning new skills to be an active participant in our evolving health care system. PMID- 11917300 TI - Shingles update: common questions in caring for a patient with shingles. AB - Varicella zoster virus (VZV) is a herpes virus that can cause two distinct clinical diseases, chickenpox and shingles. Primary infection of varicella, often called chickenpox, results in a generalized eruption of a vesicular exanthematous rash which is usually seen in children and is highly contagious. This virus (VZV) can then become latent and later reactivate causing herpes zoster, commonly known as shingles. Shingles is usually a localized phenomenon often seen in adults and is usually less contagious. The following is a discussion of infection control questions most commonly asked regarding the care of a patient with shingles. PMID- 11917302 TI - Melanoma update. PMID- 11917301 TI - Wound assessment and evaluation. PMID- 11917303 TI - Acne news. PMID- 11917304 TI - Yellow eyelids heralding lymphoma. AB - Yellow eyelids are an uncommon finding but can be cosmetically disfiguring to a patient. There are a variety of causes of yellow eyelids. A case study as well as the differential diagnosis of yellow eyelids are presented. PMID- 11917306 TI - Skin rejuvenation with cool touch 1320 nm Nd:YAG laser: the nurse's role. AB - Nonablative laser remodeling with the Cool Touch system is a new skin rejuvenation procedure. Nurses play several valuable roles in educating and caring for patients undergoing this procedure including assisting with patient selection, preoperative preparation, intraoperative set up and delivery, postoperative care, and followup. PMID- 11917305 TI - Disposable baby wipes: efficacy and skin mildness. AB - The results of a series of four clinical studies demonstrated that disposable baby wipes were milder to the skin than use of a cotton washcloth and water, recognized as a "gold standard" for skin mildness. Importantly, the baby wipes caused no significant change from the baseline value in any of the skin parameters examined. This observation verified that the test wipes are minimally disruptive to the epidermal barrier and thus suitable for use on intact or compromised, irritated skin. PMID- 11917307 TI - The leader as synergist. PMID- 11917308 TI - What's your assessment? Psoriasis. PMID- 11917311 TI - Wound assessment and evaluation: documentation of pressure ulcers. PMID- 11917312 TI - Shaping the system and culture of health care. PMID- 11917313 TI - Treatments for common superficial fungal infections. AB - Fungal infection of keratinized tissue is caused by any of the dermatophyte species. The topical allylamines and benzylamines have been especially effective in treating these infections because of their in vitro fungicidal activity and short treatment duration. With the development of new oral azoles and allylamine antifungal agents, there has been a renewed interest in treating superficial skin fungal infections. The use of topical and oral antifungal agents in treating cutaneous fungal infections is examined. PMID- 11917314 TI - Estimating true burden of disease detected by screening tests of varying validity. AB - Timely and accurate information on disease load is essential for planning health programs. Unfortunately, complexity, cost and need of skilled personnel limit the use of screening tools of high validity in developing countries. The disease load estimated with tools of low validity differs considerably from true disease load, particularly for diseases of extreme levels of prevalence/incidence. A tool of 70% sensitivity and specificity may yield a prevalence/incidence rate of 34% (CI: 32.23-35.67%) for a disease whose true rate is only 10.0% (CI: 8.94-11.06%). We proposed a procedure to derive the true estimate in such cases, based on the concepts of sensitivity and specificity of a diagnostic/screening test. It is applied on two sets of real data--one pertaining to incidence rate of low birth weight (LBW) and the other to prevalence rate of obesity--where multiple screening tests of varying validity were used to estimate the magnitude. Different screening tests yielded widely varying incidence/prevalence rates of LBW/obesity. The prevalence/incidence rates derived by using the proposed estimation procedure are similar and close to the true estimate obtained by screening tests considered as gold standard. Further, sample size determined on the basis of the results of a tool of low validity may be either larger or smaller than the required sample size. Estimation of true disease load enables determination of correct sample size, thus improving the precision of the estimate and, in some instances, reducing the cost of investigation. PMID- 11917315 TI - Declining trend in routine UIP coverage. AB - Routine UIP coverage status in the state of West Bengal and three selected Municipal Corporation areas (Calcutta, Howrah and Siliguri) were studied during 1997-98 and 1998-99. Also, UIP coverage status in the 'high risk' areas of the State (areas which reported Polio cases during 1998) was studied during 1998-99. UIP coverage in the state of West Bengal was only 54.3% in 1997-98, which further declined to 48.1% in 1998-99. In the three urban areas, UIP coverage ranged between 57.3%-70.9% in 1997-98, which further declined to 29.6%-47.1% in 1998-99. Antigenwise coverage revealed very poor performance with DPT3, OPV3, and Measles in 1997-98 and further decline in 1998-99. Dropout rate was also very high. In 1998-99 drop-out rate ranged between 30.1% to 54.2% in different studied areas. Some other studies suggested that PPI activities, which are very visible and targetted programme, may adversely affect routine UIP services. There is urgent need for further probing to identify the reasons for such poor state of affairs, keeping PPI angle in mind and to initiate remedial measure urgently. PMID- 11917316 TI - Prevalence of anaemia among adolescent girls in rural area of District Meerut, U.P. AB - The prevalence of anaemia in 504 adolescent girls (10-18 yrs) representing 24 subcentre villages of Daurala block of Meerut was 34.5%. The prevalence of mild, moderate and severe anaemia among adolescent girls was 19.0%, 14.0% and 1.4% respectively. Majority (55.2%) were having mild anaemia and only 4.0% had severe anaemia. Anaemia was found to be significantly associated with educational status (P < 0.05), birth order (P < 0.05), awareness regarding anaemia (P < 0.05) and marital & obstetric status (P < 0.05) with no association with age, anthropometry and menarcheal age (P > 0.05). PMID- 11917318 TI - Adolescent health. PMID- 11917319 TI - Malariogenic situations in areas of Aiodhya hills of the district Purulia West Bengal and its present status. PMID- 11917317 TI - A study on awareness of AIDS among school students and teachers of higher secondary schools in north Calcutta. AB - Higher Secondary School students and their teachers were studied to assess the knowledge about AIDS and attitude towards AIDS patients. Only 13.5% senior school students and 16.2% teachers had clear knowledge regarding AIDS--its general aspects, transmission and prevention. Girls had higher and clear knowledge than boys. 45.8% of girls, 38.8% of boys students and 20.3% of teachers had positive attitudes towards nursing an AIDS case. It is suggested that schools have to device ways to open up more effective communication with students in relation to education on sex and AIDS. Training on AIDS should be emphasized on school teachers who on their turn can teach the students in a correct way about AIDS. PMID- 11917320 TI - Reproductive tract infection in women of reproductive age group in Sitapur/Shahjahanpur District of Uttar Pradesh. AB - CARE India with its UP Office has initiated demand generation and acceptability of services for reproductive health and birth spacing services at community level in two districts in the state of Uttar Pradesh. A total of 3,49,000 women of reproductive age will benefit from participation in program activities over the next five years. Out of total surveyed population of 3662, 798 women of reproductive age group were identified of whom 272 were found to be symptomatically positive in relation of STDs/RTIs giving the overall prevalence rate of 34%. Clinical as well as the microbiological examination was carried out on 193 of these 272 women in field conditions. Those found clinically and/or microbiologically positive were provided treatment and preventive advice. Total of 150 (77.7%) women were diagnosed on the basis of symptoms (syndromic approach) and 151 (78.2%) were diagnosed on the basis of clinical examinations. However, 137 (70.9%) were confirmed microbiologically positive. A large extent was found in the age group of 15 to 24 years. Confirmed cases were comparatively less among nulliparous women (63.6%) than multipara (73.2%) cases. 64.1% women were having watery discharge while 29.1% were having complaint of curd discharge. The discharge was offensive in only 1.9% cases while 4.9% reported mixed discharge. 54.4% women complaining of vaginal discharge were suffering from PID. The discharge were either bacterial alone (26.1%) or bacterial mixed with candidiasis (16.5%) or trichomoniasis (8.7%). Compared to syndromic approach, an improvement in the sensitivity (81.8%) and predictive accuracy (74.1%) was noted when cases were detected with the help of clinical examination. PMID- 11917321 TI - The missing millions: a sad commentary on gender discrimination. PMID- 11917322 TI - Kala-azar: a potentially eradicable disease as a public health challenge. PMID- 11917323 TI - Rural people and fine needle aspiration cytology (FNAC)--some findings in North Bengal Medical College, Darjeeling, West Bengal. AB - A hospital based interdepartmental collaborative study was carried out from 1st July to 30th September, 2000 on 269 rural people residing in six districts of the northern part of West Bengal and attending the Pathology Department in North Bengal Medical College for Fine Needle Aspiration Cytology (FNAC) after being referred from different clinical departments. The objectives of the study were to study some aspects of the outcomes of FNAC among rural people and to suggest for its wider application in rural community through Community Health Centre/Rural Hospital/Block Primary Health Centre. The results shows that Hindus (80.3%) are attending more in number than Muslims (14.13%), Christians (4.83%) and Buddhist (0.74%); 53.54% of the population are in 11-40 years age group. In benign conditions 76.95% are coming from up to 100 kms. of N.B.M.C; people in lower per capita income group of up to Rs.400/- per month are attending twice in number than those in the income groups of Rs.401/- and more per month. In malignancy however people do not think of distance or expenses due to seriousness of the diseases. In 14.5% cases FNAC remains inconclusive whereas in 85.5% cases it provides definite diagnoses. PMID- 11917324 TI - Effectiveness of training on infant feeding practices among community influencers in a rural area of west Bengal. AB - Total 34 Influencers were trained in a subcentre area of South 24-parganas district of West Bengal. Knowledge was imparted to community influencers on infant feeding practices through lecture, group discussion, question-answer session and hand-on-training by trained health workers. Pre-assessment was done before initiation of training. Repeat training was conducted at frequent intervals within a period of 3 months. Mean score of knowledge of influencers during pre-training assessment was 13.3 and improved thereafter-following training to 20.8 (1st assessment), 20.6 (2nd assessment), 23.7 (3rd assessment) and 25.2 (final-assessment). Repeat training had also desired impact. PMID- 11917325 TI - Tobacco: the addictive slow poison. PMID- 11917326 TI - Patterns of tobacco use amongst school teachers. AB - Indulgence in tobacco is a known health risk. The example of school teachers in this regard is likely to have far reaching influence on their pupils and the community. A cross sectional study among 257 teachers from 30 schools revealed that as high as 51.0% teachers consumed tobacco in one form or the other. Smoking was the most popular form of tobacco use (72%). Marked gender differences were noted with 73.9%. Male teachers hooked to tobacco habit in comparison to 13.9% of female teachers. Educational qualification had paradoxical effect on tobacco habit as fewer (20.0%) graduate teachers used tobacco in comparison to non graduate teachers (55.7%). However proportion of Post graduate teachers consuming tobacco was still higher (64.2%). Initiation to tobacco habit in majority began at age 21 years and beyond. Common reasons given for tobacco consumption were curiosity (37.9%), to be social (22.0%), enjoyment (21.2%), to relieve stress (8.17%) and improving performance (5.8%). 21.9% smokers consumed more than 20 cigarettes a day. More than four fifth teachers consciously avoided tobacco use in school premises. Nearly half (45.3%) of tobacco users reported reduction in tobacco use in preceding two years. By and large teachers (92.4%) were aware of harmful consequences of tobacco on health. But only in 29.6% it was a cause for concern. 71.2% respondents intended to give up tobacco habit. 33.7% smokers were aware of hazards of passive smoking to others. PMID- 11917327 TI - Impact of modified leprosy elimination campaign in a MDT pilot project district of India. AB - A study was carried out based upon the data available from National Leprosy Eradication Program of Purulia district in West Bengal. The result showed that the disease was in a declining trend up to the year 1998 and was inclining during the year 1998-1999 due to more case detection through MLEC-I, then followed by declining trend during the year 2000 as it was observed by MLEC-II. Single skin lesion rate was higher, MB rate was same and reduced deformity rate indicates early detection of cases due to better awareness of the community about the disease. PMID- 11917328 TI - Socio-economic factors associated with malaria in a tribal area of Orissa, India. AB - Study on the socio-economic factors and human behaviour in a cross-section of tribal communities in Sundargarh district, Orissa revealed that poor socioeconomic status and socio-cultural factors play important role in maintaining high degree of malaria transmission. Human behaviour such as location of hamlets, type of housing, sleeping habits, outdoor activities after dusk, poor knowledge about the disease and treatment seeking behaviour are of great significance as determinants of malaria transmission. All these factors need to be considered before planning community health programme. Estimation of economic loss due to malaria showed an average loss of 8.96 mandays per malaria patient with an average loss of 3.84 mandays to other family members. Mean total loss per malaria episode comes to Rs.334.91. The study showed that malaria is one of the major disease affecting the tribals to the greatest extent and putting a lot of burden on the economic upliftment of these communities. PMID- 11917329 TI - Monitoring and continuing education system of ICDS Programme in Hooghly District, West Bengal. AB - The functioning of "Monitoring and Continuing Education System" of ICDS Programme in Hooghly District was studied through record analysis and individual interviews of Anganwadi Workers (AWW). No Sector Adviser was entrusted the task of monitoring and continuing education activities in 15 out of 17 sectors, supervisors and Health Workers were present in 88.0% and 29.4% sector meetings respectively. The sector meetings were not usually held on fixed dates. Visits of Anganwadi centres (AWC) by Health Workers, Supervisors and Child Development Project Officers (CDPO) were very infrequent and no joint visit was made. Only 11.8% Anganwadi Workers were exposed to continuing education sessions. There was gross under reporting of pregnant mothers and live births. Need of all-round improvement of functioning of 'Monitoring and Continuing Education System' of ICDS Programme is well felt. PMID- 11917330 TI - Promoting health, changing behaviour--issues and challenges. PMID- 11917331 TI - Study of interspouse communication and adoption of family planning and immunization services in a rural block of Varanasi District. AB - Interspouse communication was studied in some pertinent areas which have an important bearing on day to day transactions. The level of such communication measured on a three-point scale was studied for its role on acceptance of family planning and immunization services. 200 currently married females residing with their husbands in a rural block of Varanasi, in the reproductive age group, with at least one child aged 1-3 years were selected and interviewed. Scores were ascribed for 12 selected items of conversation according to frequency of conversation on a three point scale based on which high, medium and low communicators were delineated. In this study high, medium and low communicators were found to be 14%, 40% and 45% respectively. Topics of importance which never featured in interspouse communication were menstrual problems (44%), when to have first child (82.5%) and birth spacing (48.5%). Interspouse communication was better in upper castes and joint families. Literacy status of both husband and wife and per capita income of the family revealed positive relationship with inter-spouse communication. Adoption and practice of family planning methods as well as full immunization coverage of the child in the family were observed to be higher among high and medium communicators as compared to low degree of communcators (p < 0.001). PMID- 11917333 TI - Risk factors of diarrhoea among flood victims: a controlled epidemiological study. AB - The concept and practice of 'disaster preparedness and response', instead of traditional casualty relief, is relatively new. Vulnerability analysis and health risks assessment of disaster prone communities are important prerequisites of meaningful preparedness and effective response against any calamity. In this community based study, the risk of diarrhoeal disease and its related epidemiological factors were analysed by collecting data from two selected flood prone block of Midnapur district of West Bengal. The information was compared with that of another population living in two non-flood prone blocks of the same district. The study showed that diarrhoeal disease was the commonest morbidity in flood prone population. Some behaviours, like use of pond water for utensil wash and kitchen purpose, hand washing after defecation without soap, improper hand washing before eating, open field defecation, storage of drinking water in wide mouth vessels etc. were found to be associated with high attack rate of diarrhoea, in both study and control population during flood season compared to pre-flood season. Attack rates were also significantly higher in flood prone population than that of population in non-flood prone area during the same season. Necessity of both community education for proper water use behaviour and personal hygiene along with ensuring safe water and sanitation facilities of flood affected communities were emphasized. PMID- 11917332 TI - Coverage of Maternal Care Services in the state of West Bengal. AB - Evaluation of the Coverage of Maternal Care Services was conducted during the year 1999 in the entire State of West Bengal, Urban poor localities of randomly selected three Municipal Corporation areas and 'high-risk' urban wards or villages of West Bengal that had reported AFP cases in 1998. It was observed that mothers who had three or more antenatal check-ups varied between 54% to 82% in different study areas. Tetanus Toxoid coverage varied between 83.5% to 93.4% being lowest in high-risk areas. Only a very small section of mothers (12.7% to 23.7%) consumed 100 or more Iron and Folic acid tablets during pregnancy. 'Danger Signs' of pregnancy were informed to only 21.1% to 38.2% of the pregnant mothers. Proportion of home deliveries varied between 16.7% (Calcutta) to as high as 72.7% (high-risk areas). Deliveries conducted by untrained personnel were 60.8% in high risk areas, 38.1% in State clusters and between 14.2% to 29.4% in the three urban areas. Vast majority of mothers (78.8% to 88.4%) received no post-natal check-up. PMID- 11917334 TI - Nutritional status of adolescent girls of a slum community of Varanasi. AB - In order to assess nutritional status of adolescent girls of a slum community of Varanasi and factors influencing them, this study was carried out on 70 girls belonging to the age group 13 to 18 years. The study subjects were selected from Sunderpur, an urban community of Varanasi, by adopting appropriate sampling methodology. The approach adopted for the study was a cross sectional one. The tools in the study were pre-designed and pre-tested schedule, weighing scale, steel anthoropometric rod and measuring tape. The techniques of the study included interview method, clinical examination and anthropometry. In all, 70.0% adolescent girls had BMI < 20%; 51.43% study subjects were suffering from Chronic Energy Deficiency (CED). Stunting (Height for age alpha 90%) was present in 10% of adolescent girls. Their average Weight, Height, MAC were 83.45%, 93.08% and 82.05% of the corresponding estimated reference values. Significant association of common parameters (viz., age, caste, income, type of family, working and literacy status) with nutritional status of study subjects was not observed in this study. However, lesser undernutrition in large families (> 6) indicated role of familial support in prevention of undernutrition in adolescent girls. PMID- 11917335 TI - Hepatitis B viral infection amongst hospital personnel in Calcutta. PMID- 11917337 TI - Using the National Guideline Clearinghouse. PMID- 11917336 TI - You've probably got guys like this in your shop. So you should know.... Why people worry at work. PMID- 11917338 TI - Internal and external consulting. Assisting clients with managing work, health, and psychosocial issues. AB - 1. Consulting is the art of influencing people at their request. It means reaching a conclusion of acceptable quality, maintaining an acceptable commitment to that conclusion, supporting the client to prevent or minimize any detrimental outcomes as a result of the conclusion, and being perceived as helpful and efficient by the client. 2. Occupational and environmental health nurses are involved in a variety of consulting roles along with different types of consulting relationships, including both internal and external consulting. 3. To be an effective consultant, occupational and environmental health nurses need specialized knowledge and skills. Effective consultants work hard at developing sensitivity and competency to help others, and leave them self sufficient when the project is complete. PMID- 11917339 TI - Effectiveness of installing overhead ceiling lifts. Reducing musculoskeletal injuries in an extended care hospital unit. AB - The effectiveness of replacing floor lifts with mechanical ceiling lifts was evaluated in the extended care unit of a British Columbia hospital. Sixty-five ceiling lifts were installed between April and August 1998. Injury data were abstracted from injury reports for all staff musculoskeletal injuries (MSI) occurring in the unit during a 3 year period prior to installation and a 1.5 year follow up period. Descriptive statistics were calculated for injuries pre- versus post-installation. Rates were calculated as number of injuries per 100,000 worked hours. Rates for three pre- and three post-installation intervals were compared using Poisson regression. The rate of MSI caused by lifting/transferring patients was significantly reduced (58% reduction, p = .011) after installation, but rates of all MSI and MSI caused by repositioning did not statistically decline (p > .05). Further follow up is necessary to determine whether or not ceiling lifts also can be effective for decreasing injuries related to repositioning patients on this unit. PMID- 11917340 TI - Implementing a resident lifting system in an extended care hospital. Demonstrating cost-benefit. AB - 1. Implemeting mechanical resident lifting equipment in an extended care facility produced a payback from direct savings alone within 4 years. Payback occurred more quickly when the effect of indirect savings or the trend to rising compensation costs was considered. 2. Combining the observations of the occupational health nurses related to staff well being with relevant cost-benefit data is useful in influencing decision makers and in securing funding for prevention measures. 3. Clear identification of a viewpoint is an important part of an economic evaluation and cost-benefit analysis. PMID- 11917341 TI - Mentoring. Career enhancement for occupational and environmental health nurses. AB - Mentoring is a career development and enhancement tool for occupational and environmental health nurses. A history of mentoring in the literature has provided the foundation for understanding mentoring, including the traditional and newer types of mentoring relationships. The benefits and barriers to mentoring for occupational and environmental health nurses include access and mentor-protege qualities. Using a multiple mentor approach in which mentoring relationships change with career development, placement, and needs, rather than a traditional approach where one mentoring relationship is established early in the career and lasts throughout the career, may be most beneficial to occupational and environmental health nurses. PMID- 11917342 TI - [Cross section NuclearMedicine]. PMID- 11917343 TI - Combined image interpretation of computed tomography and hybrid PET in head and neck cancer. AB - AIM: Evaluation of potential synergistic effects of combined image interpretation of FDG PET using a gamma camera modified for coincidence detection (hybrid PET) and computed tomography (CT) and comparison of the diagnostic accuracy of hybrid PET and dedicated PET in patients with head and neck cancer. METHODS: Forty-two patients with suspected primary or recurrent cancer were included. Twenty-four patients underwent dedicated PET in addition to attenuation-corrected hybrid PET using a one-day protocol. RESULTS: Sensitivity, specificity and accuracy for detection of primary or recurrent head and neck cancer were 74, 73, and 74% for hybrid PET, 52, 82, and 60% for CT and 77, 82, and 79% for combined reading. With the combination of CT and hybrid PET all cases of recurrent disease were detected. The largest tumour not detected was 1.7 cm in diameter. Sensitivity, specificity and accuracy for the detection of neck sides with lymph node metastases were 69, 88, and 85% for hybrid PET, 62, 88, and 84% for CT, 69, 99, and 94% for combined image interpretation. With combined interpretation four involved neck sides were missed including two cases of microscopic metastases. Hybrid PET revealed concordant results to dedicated PET in all patients with respect to the detection of primary or recurrent tumour and in 45 of 48 neck sides (94%) with the same number of false negative findings. CONCLUSION: The combination of functional information of hybrid PET and morphological information of CT by the simple approach of combined image interpretation improves the sensitivity for the detection of primary/recurrent head and neck cancer and increases the specificity of lymph node staging compared to CT alone. The accuracy of hybrid PET and dedicated PET was almost identical. PMID- 11917344 TI - [F-18-FDG hybrid camera PET in patients with postoperative fever]. AB - AIM: Evaluation of F-18-FDG-hybrid-camera-PET imaging in patients with undetermined postoperative fever (POF). METHODS: Prospective study of 18 patients (9 women, 9 men; age 23-85 years) suffering from POF with 2-fluoro-2' deoxyglucose (F-18-FDG) using a dual headed coincidence camera (DHCC). Surgery had been performed 5-94 days prior to our investigation. 13 of the 18 patients received antibiotic therapy during the time of evaluation. Ten (55%) had an infectious and eight (45%) a non infectious cause of fever. RESULTS: Increased F 18-FDG-uptake outside the surgical wound occurred in 13 regions (infection n = 11, malignancy n = 2). The sensitivity of F-18-FDG-hybrid-camera-PET in imaging infection in areas outside the surgical wound was 86% and the specificity 100%, respectively. Antibiotic therapy did not negatively influence the results of F-18 FDG-scanning. Increased F-18-FDG-uptake within the surgical wound was seen in 8 of 18 patients. The sensitivity of F-18-FDG-hybrid-camera-PET in imaging infection within the surgical wound was 100% and the specificity 56%, respectively. The interval between surgery and F-18-FDG-scanning was significantly shorter in patients with false positive results compared with patients showing true negative results (median 34 vs. 54 days; p = 0.038). CONCLUSION: In POF-Patients, F-18-FDG transaxial tomography performed with a F-18 FDG-hybrid-camera-PET is sensitive in the diagnosis of inflammation and malignant disease within and outside the surgical wound. Because of the accumulation of the tracer both in granulation tissue and infection, the specificity in detecting the focus of fever within the surgical wound is poor. PMID- 11917345 TI - [Methods and clinical applications in nuclear cardiology: a position statement]. AB - Nuclear cardiological procedures have paved the way for non-invasive diagnostics of various partial functions of the heart. Many of these functions cannot be visualised for diagnosis by any other method (e.g. innervation). These techniques supplement morphological diagnosis with regard to treatment planning and monitoring. Furthermore, they possess considerable prognostic relevance, an increasingly important issue in clinical medicine today, not least in view of the cost-benefit ratio. Our current understanding shows that effective, targeted nuclear cardiology diagnosis--in particular for high-risk patients--can contribute toward cost savings while improving the quality of diagnostic and therapeutic measures. In the future, nuclear cardiology will have to withstand mounting competition from other imaging techniques (magnetic resonance imaging, electron beam tomography, multislice computed tomography). The continuing development of these methods increasingly enables measurement of functional aspects of the heart. Nuclear radiology methods will probably develop in the direction of molecular imaging. PMID- 11917346 TI - [Tc-99m ciprofloxacin in clinically selected patients for peripheral osteomyelitis, spondylodiscitis and fever of unknown origin--preliminary results]. AB - AIM: Retrospective evaluation of Tc-99m ciprofloxacin (infection) scintigraphy consecutively performed in a series of patients clinically suspected for peripheral osteomyelitis (OM), spondylodiscitis (SD) and fever of unknown origin (FUO). METHODS: A total of 20 patients clinically suspected for OM (n = 12), SD (n = 3) and FUO (n = 5) were included in our retrospective analysis. The additional criterion was a positive 3-phase bone scan for OM, or a 2-phase bone scan in case of SD. Planar whole body scans and static acquisitions were performed 1 and 4 h after application of 370 MBq Tc-99m ciprofloxacin. In 10 patients with suspected OM, additional immunoscintigraphy using Tc-99m labelled monoclonal antibodies (Mab BW 250/183) was performed and the correlation of infection to bloodpool and antigranulocyte scintigraphy was analysed. RESULTS: OM: Bacterial infection was confirmed in 8 of 15 lesions. Infection demonstrated true positive (TP) results in 7 of 8, true negative (TN) results in 2 of 7, false positive (FP) results in 5 of 7 patients and one false negative (FN) result. A strong correlation could be demonstrated between T/NT ratios of infection and bloodpool Tc-99m medronate imaging (r = 0.84, 0.88) and between infection and BW 250/183 (r = 0.92, 0.90). Using a threshold of 2.0 for T/NT ratio, only TP results could be observed whereas a T/NT in the range of 1.0-2.0 could not discriminate between septic and aseptic inflammation. Concordant results with Mab BW 250/183 could only be observed in 5 of 10 patients (4 TP, 1 TN) by showing 4 FP and 1 FN lesions with IF. CONCLUSION: Non-specific uptake of infection can be observed in a variety of clinical situations with moderate uptake, by showing a strong correlation with blood-pool imaging. Nevertheless, intense uptake may be specific for septic inflammation. PMID- 11917347 TI - Improving the diagnosis of acute appendicitis with atypical findings by Tc-99m HMPAO leukocyte scan. AB - AIM: Even with careful observation, the overall false-positive rate of laparotomy remains 10-15% when acute appendicitis was suspected. Therefore, the clinical efficacy of Tc-99m HMPAO labeled leukocyte (TC-WBC) scan for the diagnosis of acute appendicitis in patients presenting with atypical clinical findings is assessed. PATIENTS AND METHODS: Eighty patients presenting with acute abdominal pain and possible acute appendicitis but atypical findings were included in this study. After intravenous injection of TC-WBC, serial anterior abdominal/pelvic images at 30, 60, 120 and 240 min with 800 k counts were obtained with a gamma camera. Any abnormal localization of radioactivity in the right lower quadrant of the abdomen, equal to or greater than bone marrow activity, was considered as a positive scan. RESULTS: 36 out of 49 patients showing positive TC-WBC scans received appendectomy. They all proved to have positive pathological findings. Five positive TC-WBC were not related to acute appendicitis, because of other pathological lesions. Eight patients were not operated and clinical follow-up after one month revealed no acute abdominal condition. Three of 31 patients with negative TC-WBC scans received appendectomy. They also presented positive pathological findings. The remaining 28 patients did not receive operations and revealed no evidence of appendicitis after at least one month of follow-up. The overall sensitivity, specificity, accuracy, positive and negative predictive values for TC-WBC scan to diagnose acute appendicitis were 92, 78, 86, 82, and 90%, respectively. CONCLUSION: TC-WBC scan provides a rapid and highly accurate method for the diagnosis of acute appendicitis in patients with equivocal clinical examination. It proved useful in reducing the false-positive rate of laparotomy and shortens the time necessary for clinical observation. PMID- 11917348 TI - Moyamoya syndrome: impaired hemodynamics on ECD SPECT after EEG controlled hyperventilation. AB - BACKGROUND AND PURPOSE: Ischemic symptoms in children with Moyamoya syndrome are typically provoked by hyperventilation (HV) and are accompanied by the "re-build up" phenomenon in EEG. The value of scintigraphic detection of HV-provoked perfusion deficits remains to be elucidated. PATIENTS AND METHODS: In seven children with Moyamoya syndrome regional cerebral blood flow was assessed by 99mTc-ethyl-cysteine-dimer (ECD) single photon emission computed tomography (SPECT) after HV and under baseline conditions to identify ischemia prone regions. RESULTS: Regional marked hypoperfusion after HV was found in all patients. Predominant perfusion deficits were detected in the frontal lobes. CONCLUSION: ECD SPECT is a potential tool for the preoperative evaluation of cerebral hemodynamics and for monitoring angiosurgical therapies in Moyamoya disease. PMID- 11917349 TI - [Body weight gain after radioiodine therapy in hyperthyroidism]. AB - AIM: Analysis and follow up of body weight after radioiodine therapy (RITh) of hyperthyroidism, since excessive weight gain is a common complaint among these patients. METHODS: Therapy and body weight related data of 100 consecutive RITh patients were retrospectively analysed from the time before up to three years after RITh. All patients suffered from hyperthyroidism (Graves' disease or autonomy), but were adjusted to euthyroid levels after RITh. Patients' data were compared to a control group of 48 euthyroid patients out of the same ambulance and during the same time scale. RESULTS: All patients (RITh and controls) gained weight over the time. There was no statistically significant difference in BMI development over three years between RITh-patients and controls (5.5% resp. 4.9% increase). In the first year after RITh, weight gain of the RITh patients was higher indeed, but lower in the follow up, resulting in the same range of weight gain after three years as the controls. Besides that women showed a slightly higher increase of BMI than men, and so did younger patients compared to elder as well as patients with overweight already before RITh. CONCLUSIONS: An initially distinct increase of body weight after RITh of hyperthyroidism is mainly a compensation of pretherapeutic weight loss due to hyperthyroidism. Presupposing adequate euthyroid adjustment of thyroid metabolism after therapy, RITh is not responsible for later weight gain and adipositas. PMID- 11917350 TI - [Aa DICOM based PACS for nuclear medicine]. AB - The installation of a Radiology Information System (RIS) connected to a Hospital Information System (HIS) and a Picture Archiving and Communications System (PACS) seems mandatory for a nuclear medicine department in order to guarantee a high patient throughput. With these systems a fast transmission of reports, images to the in- and out-patients' wards and private practitioners is realized. Therefore, since April 2000, at the department of nuclear medicine of the university of Wurzburg a completely DICOM based PACS has been implemented in addition to the RIS. With this system a DICOM based workflow is realized throughout the department of nuclear medicine for reporting and archiving. The PACS is connected to six gamma-cameras, a PET scanner, a bone densitometry system and an ultrasound device. The volume of image data archived per month is 4 GByte. Patient demographics are provided to the modalities via DICOM-Worklist. With these PACS components a department specific archive purely based on DICOM can be realized. During the installation process problems occurred mainly because of the complex DICOM standard for nuclear medicine. Related to that is the problem that most of the software implementations still contain bugs or are not adopted to the needs of a nuclear medicine department (particularly for PET). A communication software for the distribution of nuclear medicine reports and images based on techniques used for the worldwide web is currently tested. PMID- 11917351 TI - [Subcutaneous splenosis--demonstration with colloidal 99m-Tc-rhenium sulfide]. PMID- 11917352 TI - [12. Thyroid gland discussion. Therapy for hyper-and hypothyroidism]. PMID- 11917353 TI - [Therapy of thyroid cancer. Recombinant TSH]. PMID- 11917354 TI - [Multiple pregnancy and tocolysis]. PMID- 11917355 TI - [Safety of patient transport for in utero fetal indication]. PMID- 11917356 TI - [Prophylactic ambulatory treatment for premature labor (with the exception of premature labor with rupture of the membranes)]. PMID- 11917357 TI - [Diagnosis of premature labor]. PMID- 11917358 TI - [Treatment of premature labor]. PMID- 11917359 TI - [Obstetric ultrasound in danger]. PMID- 11917360 TI - [Indicators and evaluation tools for perinatal care networks]. AB - Since the regionalization of perinatal services and the laws on the safety of child-birth were announced on the 9th October 1998, conventions and rules establishing relationships between doctors in perinatal networks have gradually been set up. In order to assess the impact, functioning and effectiveness of this care organisation on the health of mothers an childs, a common study design is essential. Such a common protocol, with the same methodology, allows the comparison of observed results from one perinatal network to another. With this objective, four leading societies in the field, the French National College of Gynaecologists and Obstetricians (CNGOF), the National Federation of Neonatal Pediatricians (FNPN), the French Society of Perinatal Medicine (SFPM) and the Association of Computerised Medical Records in Perinatalogy, Obstetrics and Gynaecology (AUDIPOG) set up a workshop in order to elaborate a common text, which was adopted by the Administrative Councils of the four societies. This contractual text, co-operatively elaborated, is now introduced to the scientific community. It was prepared from scientific papers and international recommendations in the field of perinatal regionalization, leading to a protocol giving information on objectives, study population, study design and assessment criteria. For this, a list of questions was proposed, including the impact of perinatal network, its acceptability by doctors and families, its functioning and its effectiveness on the health of mother and child. The means of data collection were then presented, whether from institutional sources of perinatal data (completed from specific registers or studies), or from the information system France-Perinat, especially designed for the evaluation of the perinatal networks. PMID- 11917362 TI - [Influence of the type of breech presentation on delivery in selected primiparous women at term]. AB - OBJECTIVE: Study the effect of the type of breech presentation on the prognosis of delivery in selected primiparous women at term. PATIENTS AND METHODS: We reviewed retrospectively 146 breech presentations seen between January 1, 1993 and December 12, 1999 in our unit. We limited our study to primiparous mothers at term (> or = 37 weeks gestation) who had had a normal single fetus pregnancy and who had accepted a trial of labor. We analyzed the mode of labor termination, and maternal, fetal, ovular, obstetrical and neonatal features by type of breech presentation and by type of delivery. We collected data by manual search of the patients' registered files (AUDIPOG). The chi-squared and Fisher's tests were used as appropriate for statistical analysis. RESULTS: Spontaneous vaginal delivery was achieved in 10.3% of the cases and non-spontaneous vaginal delivery in 56.2%. Cesarean section during labor was required in 33.5%. There was no significant difference between frank breech presentation (27.4%) and incomplete breech presentation (72.6%). Maternal, fetal, ovular, obstetrical and neonatal features were not significantly different. CONCLUSIONS: Unlike data reported by others, our series did not demonstrate any significant feature predictive of obstetrical and neonatal prognosis irrespective of the type of breech presentation in selected primiparous women at term who had had a normal pregnancy. PMID- 11917361 TI - [GnRH antagonists in IVF]. AB - New GnRH antagonists are available in clinical practice. The different studies have confirmed the efficacy of these antagonists in preventing the LH surge. Two protocols have been described: in the multiple dose regimens, small doses of antagonist (0.25 mg) are injected starting on stimulation day 5 or 6 until hCG. In the single dose protocol, one injection of a larger dose (3 mg) is proposed in the late follicular phase. Local and general tolerance of the two compounds is very good. The results obtained with both regimens as compared with GnRH agonists in long protocols are showing a reduction in the stimulation length, in the consumption of gonadotrophins and in the incidence of the OHSS. The pregnancy rates are comparable in the good prognosis patients selected in the published studies. When the final tuning of these new protocols will be done, the advantages of GnRH antagonists in reducing the complications and side effects of ovarian stimulation will give to GnRH antagonists an important place in IVF. PMID- 11917363 TI - [Oxytocic use and duration of labor under peridural ambulatory analgesia]. AB - OBJECTIVE: Extradural analgesia is associated with an increase duration of labor and oxytocin use. MATERIAL AND METHODS: We conducted a study of walking during active labor under extradural analgesia among 41 women. This group, with uncomplicated pregnancies, was similar to associate with a usual-care group according to the following criteria: maternal age, parity, gestational age, Bishop's score at the admission, and labor entrance mode. Analgesia was performed by using combined spinal-epidural analgesia and patient controlled extradural analgesia. Fetal heart and uterine contractions were continuously monitored using Telemetry. Anesthetic and obstetrical parameters were noted every hour. The main criteria were length of labor and oxytocin consumption. Statistical analysis was performed using Student's t-test and Fischer's exact test. RESULTS: Duration of the 1st and 2nd stage of labor, and oxytocin consumption were not statistically different in the two groups, neither was mode of delivery. No neonates had an Apgar score less than 5 at one minute. Parturients walked about 2 h 30 for about 2,000 m. CONCLUSION: Obstetrical parameters and oxytocin consumption are not modified during walking extradural analgesia. No maternal or neonatal adverse effects were noted in our study. PMID- 11917364 TI - [Birthing centers: review of the literature]. AB - Birth Centers have been established in many industrialized countries but no facilities of this type are available in France. The aim of this review was to analyze the literature concerning existing Birth Centers in order to compare them with conventional hospitals in terms of efficacy. Nine studies were retained including one prospective study. All underlined an improvement in both maternal and perinatal morbidity and mortality in the low-risk pregnancy group. No conclusion can however be made concerning the harmlessness of establishing Birth Centers in the French healthcare system. The small number of cases in these studies, their heterogenicity and the fact that only one was prospective point out the importance of cautious interpretation. Estimation studies must be conducted in experimental sites before opening Birth Centers in France. PMID- 11917365 TI - [For or against birthing centers: a survey of practitioners in the Isere area]. AB - A questionnaire survey on the Birth Centers project was conducted among gynecology and obstetrics healthcare workers in the Isere area. The aim was to better understand the organization of the healthcare system operating around childbirth and to gather suggestions and criticisms concerning this project. Among the 451 people questioned, 42% answered, including 72.6% midwives and 21% physicians. The study was conducted in three phases: exploratory talks with practitioners, drafting the 37-iem questionnaire, and analysis of the answers around nine themes: health worker's information about Birth Centers, awareness of these structures, practice of birth preparation, advice given about practical aspects of Birth Centers, midwife skills, health workers satisfaction concerning the organization of the childbirth healthcare system, ways to improve management of birth in France, healthcare workers opinion about the project and social determinants. Healthcare workers were found to be poorly informed (only 20% were aware of Birth Centers). There was also a high rate of dissatisfaction concerning the current childbirth healthcare system (78%). Most of the responders (38 to 88%) agreed with the underlying principle of Birth Centers (no induction of delivery, early discharge with home care, better information about epidural anesthesia, choice of position during labor and childbirth...), but many discrepancies were found concerning discontinuous fetal monitoring during labor and lack of epidural anesthesia in Birth Centers. Eighty percent accepted the project, but 20% though that one or several physicians should be available in the Birth Center and 47% considered the project would not affect maternal and perinatal mortality/morbidity. This constitutes a bias concerning the health workers' approval of the project. As a result, before thinking about the installation of Birth Centers in France, it would be important to evaluate the project at experimental sites and provide sufficient information to gynecology and obstetrics healthcare workers. This information must concern midwife skills and the mission of birth Centers. PMID- 11917366 TI - [Value of toxoplasma serology at delivery in women seronegative during pregnancy]. AB - Monthly serological screening of non immune pregnant women, required by law in France, is sometimes interrupted before delivery. As a consequence, late maternal infections resulting in a high risk of vertical transmission with potentially severe late sequelae are not detected. PMID- 11917367 TI - [Uterine rupture in Senegal. Results of 2 surveys taken in 1992 and 1996]. AB - OBJECTIVES: National prospective descriptive study on dystocia were conducted in Senegal in 1992 and 1996. We examined more closely the data on uterine rupture to determine trends between these two surveys. METHODS: Data were collected for all patients undergoing an obstetrical intervention between January 1st and December 31st in 1992 and in 1996 in one of the referral maternity-obstetrical surgery units in each of the 10 regions in Senegal. RESULTS: The rate of uterine rupture was 1 per 51 interventions in 1992 (1.87%) versus 1 per 53 interventions in 1996 (1.94%). This dramatic event was recurrent in 7 of the 10 regions. The patients involved were aged 25-35 years (68% in 1992 versus 70% in 1996), multiparous or grand multiparous (52% in 1992 versus 70% in 1996), illiterate (90%), with poor follow-up (less than 3 prenatal consultations in 56% of the cases), and were generally evacuated to the unit (80% in 1992 versus 97% in 1996) without medical care (55%). Maternal mortality was high (28% in 1992 versus 12% in 1996). Infant mortality was also very high (98% in 1992 versus 86.6% in 1996). Conservative treatment of uterine rupture was used increasingly: the rate of conservative suture rose from 3.3% in 1992 to 22% in 1996. CONCLUSION: Prevention is a challenge for healthcare workers, public authorities and the population in general. Health policy should be directed towards forming general practitioners in obstetrical surgery. Emergency surgery kits should be made available to improve the quality of care in all maternity units throughout the country. PMID- 11917368 TI - [Prenatal diagnosis of exencephaly at 10 weeks gestation, confirmed at 13 weeks gestation]. AB - First trimester diagnosis of the exencephaly anencephaly sequence has been reported since the introduction of transvaginal sonography, the earliest being described at 10 weeks 5 days gestation. We report the discovery of exencephaly at 10 weeks gestation. The diagnosis was confirmed by repeat ultrasonography at 13 weeks. PMID- 11917369 TI - [Acute carbon monoxide poisoning during pregnancy: 2 cases with different neonatal outcome]. AB - We report two cases of moderate maternal poisoning during the third trimester. They underwent hyperbaric oxygen therapy at 2.5 atmospheres for 90 minutes and were delivered at term. In one case the newborn presented an antenatal ischemic cerebral lesion probably due to monoxide poisoning. Pathophysiology and treatment of such accidents are discussed. PMID- 11917370 TI - [Apropos of the article: "Acute fetal pain" by G. Boog (J Gynecol Obstet Reprod 2001, 30: 393-432)]. PMID- 11917371 TI - [Ovarian cysts and pregnancy]. AB - FREQUENCY AND HISTOLOGY: In the first trimester of pregnancy, ovarian cysts are often functional without complications. After 16 weeks of gestation, frequency of ovarian cyst is reported between 0.5 and 3.0%. Histological results are identical to those observed in non pregnant women between 15 and 44 years, essentially dermoid cysts and functional cysts. Frequency of ovarian cancer is reported between 1 out of 15,000 and 1 out of 32,000 pregnancies. A surgical intervention is necessary if suspect signs are observed at ultrasonographic examination. RISK OF COMPLICATIONS: Almost all unilocular cyst with a diameter < 5 cm and persistent in second and third trimester are not associated with complications and are regressive during pregnancy; in this cases, abstention seems warranted. Conclusions are similar for dermoid cysts with diameter < 6 cm and without malignant criteria but there are a few studies on theses topics. No prospective studies are available to assess the risk of cancer or complications for cyst with diameter > 6 cm without malignant criteria. MANAGEMENT: Evacuation by puncture is not well evaluated and is not recommended during pregnancy. If an intervention is decided, laparoscopy is warranted until 16-17 weeks. After 17 weeks, laparotomy is the most evaluated method. OBSTETRICAL CONSEQUENCES: They are uncommon and caesarean section is warranted only if a cyst in the pelvis will obstruct labour. During caesarean section, removal of a cyst should be performed. PMID- 11917373 TI - [Doppler ultrasonography in the diagnosis of ovarian cysts: indications, pertinence and diagnostic criteria]. AB - To discriminate ovarian lesions is of particular importance in gynecological practice. Two main problems need answers: discrimination of benign and malignant adnexal masses and choice of the appropriate surgical treatment if necessary. Nearly 2% of the adnexal masses are ovarian carcinomas or border line tumors. It is now well established that ultrasonography is the gold standard for ovarian cyst diagnosis. The purpose of this work was to review the literature and to establish, with the evidence based medicine model, which parameters and existing diagnostic models using ultrasound and Doppler performs best in the evaluation of adnexal masses. Transvaginal sonography has demonstrated considerable advantage over conventional transabdominal sonography. However, transparietal sonography is still useful in large tumors. It is no longer reasonable to subject all patients undergoing pelvic sonography to bladder distension. Functional ovarian cyst characterization seems easy using sonography and Doppler. In case of complication, discrimination of such functional cyst may be difficult but spontaneous regression confirms usually the expectative management. Dermoid cysts and endometriomas seem to be easier to discriminate from other adnexal masses. Papillary formations on the inside of the cyst wall and masses with a non hyperechoic solid component are the most statistically significant predictors of a malignant ovarian mass. Ultrasound and morphologic parameters have a sensitivity of 80% and a specificity of 93%, that make this exam the gold standard for ovarian masses diagnosis. Another parameter is important: experienced hands with subjective evaluation seems to be one of the best ultrasound method for adnexal masses discrimination. Scoring system help differentiate benign from malignant masses (sensitivity 90%, VPP 50%). Doppler flow measurement and assessment of tumor vascularity by doppler energy increase the confidence with which a correct diagnosis is made. Moreover, combined US techniques and a diagnostic algorithm perform significantly better than morphologic assessment, color doppler or CA125 measurement alone. Logistic regression and neural network models are good methods and may be useful for malignancy prediction but the improvement is small and the concordance with histology far from 100%. In front of a benign and maybe functional cyst, spontaneous resolution may be controlled by sonographic exam at 3 and 6 months. Three-dimensional ultrasound and power doppler, contrast enhanced sonography, and sonography during the laparoscopic procedure are not still validated. Every suspicious ovarian mass needs sonography by an expert which can first use all the techniques and the different parameters to discriminate benign and malignant tumors. Secondly, after control if necessary, he can propose the patient for appropriate surgical treatment. PMID- 11917372 TI - [Histology of benign borderline ovarian and subperitoneal dystrophic cysts ]. AB - Benign cysts of the ovary include a variety of histological types. We briefly describe here the different histological types of ovarian lesions with a cystic presentation. The cystic epithelial tumors of the ovary could be definitely benign or malignant, there also exist borderline cases which are particularly difficult to manage. Close collaboration between the surgeon and the pathologist is required to carefully define indications and the limitations of frozen-section diagnosis as well as good transfer of the operative specimen. PMID- 11917375 TI - [Medical and surgical treatment of functional ovarian cysts]. AB - Most of symptomatic functional ovarian cysts (FOC) are treated by surgery. Oral contraceptives and ultrasound-guided puncture are not more efficient than expectant management for treating asymptomatic FOC in non menopausal women. The probability of a cyst being functional or benign in asymptomatic postmenopausal women is assessed by transvaginal ultrasonography, color Doppler flow imaging and CA 125 serum level measurement. Conservative management of simple cysts (anechoic, diameter < 5 cm with normal Doppler and CA 125 serum level) is more reasonable and safer than surgical treatment. The use of high-dose combination oral contraceptives has a protective effect against FOC, whereas low-dose pills have little or no effect. However, low-dose oral contraceptives or progestogen only oral contraceptives do not increase the risk of FOC. In the case of recurrent or symptomatic FOC, the advantages of using higher dose formulations have not been proved. Expectant management is as effective as oral contraceptives for the resolution of FOC induced by ovarian stimulation. Tamoxifen treatment in premenopausal breast cancer patients is associated with ovarian overstimulation and FOC formation (> 35%). Co-treatment with Gn-RH agonist may be successful in this indication. Hormone replacement therapy may be associated with an increasing ovarian volume, but not with an increasing rate of FOC. The management of surgical-induced FOC (entrapped ovarian cyst, ovarian transposition) has never been assessed by controlled studies. PMID- 11917374 TI - [Tumor markers and ovarian cysts]. AB - A practical review of tumor markers, particularly CA125 is proposed. The questions are: what is the role of CA125 assay in cancer screening programs, can preoperative CA125 level predict the benign or malignant nature of an adnexal anomaly, can postoperative CA125 predict the quality of surgical resection of cancer of the ovary? PMID- 11917376 TI - [Role of ultrasound guided puncture in the management of ovarian cysts]. AB - Ultrasound guided aspiration of ovarian cysts should be performed in well defined indications and under precise and strict modalities. Preoperative evaluation should include vaginal sonography and serum levels of CA125 and CA 19/9. Ultrasound guided aspiration is allowed only in cases of sonographically benign cyst: unilateral unilocular homogenous anechogenic cyst less than 7 cm, and serum levels of CA125 and CA 19/9 less than 35 Ul/ml. The technique of ultrasound guided aspiration is similar to the one used for oocytes recovery in in vitro fertilization treatments. It is mandatory to perform a macroscopic, cytologic and biologic (with concentrations of estradiol, CA125 and CA 19/9) evaluation of cyst fluid. Patients should be told that in case of abnormalities of cyst fluid, a laparoscopy has to be performed immediately or as soon as possible, and that there is a risk of recurrence of the ovarian lesion. PMID- 11917377 TI - [Ovarian cyst: surgical indications and access]. AB - Laparoscopic treatment of adnexal masses is indicated when all criteria of a benign lesion are present: transvaginal ultrasound demonstrates a mass < 5 cm, with liquid or dermoid content, with less than 3 fine partitions (< 3 mm), a thin wall (< 3 mm), no vegetations, normal Doppler. Laparoscopy is also indicated for "benign" cysts measuring 5 to 10 cm if laparoscopy is feasible. Peroperative exploration is the rule. Since the benign or malignant nature of an ovarian mass cannot be determined macroscopically, precaution must be taken to avoid potential laparoscopic dissemination: use of an extraction pouch, instrument cleaning, cytotoxic agent (chlorexidine or povidone-iodine) for trocar tracts, prevention of gas leakage, 3-plane suture of trocar orifices measuring > 10 mm, short interval between laparoscopic diagnosis of cancer and onset of chemotherapy or complete surgery (1 week). In case of pre- or peroperatively suspected malignancy, cytology examination of the peritoneal fluid and careful peroperative exploration of the abdomen and pelvis with peritoneal biopsy as needed are required. Simple cystectomy or adnexectomy may be performed, depending on the age of the patient, while waiting for the final pathology report. Peroperative intraperitoneal rupture must be avoided, converting to laparotomy if needed. If several suspicious elements are found, median laparotomy is often recommended, particularly in case of suspected cancer with extra-ovarian involvement, or if there is a risk of peroperative rupture. Peroperative pathology of the adnexectomy specimen and peroperative exploration will depend on the operator's experience and the availability of pathology examination in the operating room. First line laparoscopy allows an analysis of the operability and choice of the most appropriate access. PMID- 11917378 TI - [Operative technique for assumed benign ovarian cysts]. AB - Assumed benign ovarian cysts are best treated by laparoscopy rather than laparotomy (NP1). A peritoneal sampling for cytology should always be performed first (NP5). Frozen section diagnosis for suspicion of malignancy requires specific conditions (NP5). Laparoscopic treatment of macroscopically functional cysts can not be limited to needle aspiration (NP5). There is not enough evidence to privilege cystectomy after needle aspiration over attempt to achieve enucleation of the cyst; however, uncontrolled rupture of the cyst is the risk associated with the latter procedure (NP5). Authors usually recommend extraction of the cyst through a bag (NP5). There is not enough evidence for or against ovarian suture. Benefits of adhesion barriers after ovarian cystectomy has to be proved. The majority of the authors perform a bilateral oophorectomy in case of unilateral postmenopausal cyst (NP5). As much parenchyma as possible should be preserved in case of large ovarian cysts. For women with small controlateral dermoid cysts, the validity of surgical abstention is not established. Conservative surgery is the rule in case of premenopausal mucinous cysts. PMID- 11917379 TI - [Management of endometriosis ovarian cysts]. AB - The efficacy of medical treatment as unique treatment for endometrioma is not demonstrated. Operative laparoscopic management is the gold standard for surgical treatment. There is no indication to prescribe preoperatively medical treatment before cystectomy. Post-operative administration of low-dose cyclic oral contraceptive does not significantly affect long-term recurrence rate of endometriosis after surgical treatment. In cases of infertility, management of endometriomas is controversial. Recurrent ovarian surgery is not recommended. PMID- 11917380 TI - [Epidemiology of apparently benign ovarian cysts]. AB - The incidence of ovarian cysts in the general population is difficult to estimate. Certain cysts are functional and generally not operated, except in case of complications. Others are organic cysts which are usually, but not always operated. Data is available only from surgical series or pathology reports. An analysis of the literature shows that for operated cysts, approximately 75% are organic, 25% are function and 1 to 4% of the supposed benign cysts are found to be malignant. In France, the PMSI database on public and private institutions is available on the net (www.le-pmsi.fr). The number of hospital stays for benign ovarian tumors or ovarian cysts was to the order of 45,000 in 1998 and 1999. Twenty-one percent of these patients were discharged without surgery. Most had a functional cyst of the ovary. This leaves 32,000 women who underwent surgery annually for an ovarian cyst. Nearly 7000 women were hospitalized for follicular or yellow body cysts, i.e. 15%, with an incidence of 29/100,000 women over the age of 15 years. The abstention rate for hospitalized women with a follicular cyst or a yellow body cyst was 30 to 40%. For endometrioma, there were 5218 hospitalizations in 1999 (2137 in the public hospitals and 3081 in private clinics) (11% of the hospitalizations). The rate of abstention was 11% (15% in the public hospitals and 8% in private clinics). Approximately 4100 women thus underwent surgery, including 13% for ovarian cysts. According to data in the literature, serous and mucinous cysts account for 20 to 30% of the operated cysts and dermoid cysts for 10 to 20%. The type of cyst cannot be identified with the international WHO classification used in the PMSI. PMID- 11917381 TI - [Management of ovarian cysts: consequences of ovarian surgery on fertility]. AB - Surgery remains an important tool for the management of ovarian cysts. The question is whether patients of reproductive age should be given conservative treatment or undergo unilateral oophorectomy according to infertility risk. The literature is too sparse to conclude, but there are two arguments favoring conservative treatment: i) unilateral ovariectomy may impair residual ovarian function, ii) reduction of ovarian follicular number appears to be associated with increased risk of trisomy 21 in offspring. PMID- 11917382 TI - [Ovarian cysts assumed benign in the peripubertal period (10 to 16 years)]. AB - Ovarian cyst is a rare disease in infancy and childhood. Functional cysts are the most common benign ovarian tumor observed during childhood. Ultrasound examination in children with abdominal pain has improve diagnostic accuracy with an associated increase in the frequency of functional cysts (60% of the cases). Diagnosis of organic cysts is often made late. Germ cell tumors, often dermoid cysts, occur more frequently than in adults. Ultrasonography and laparoscopy are important diagnostic tools for ovarian cysts in children. Serum CEA, alpha fetoprotein and beta HGC are routine tests for organic tumors. Laparoscopy is generally proposed for treatment except for small functional cysts with a transonic structure. Spontaneous involution is generally observed. PMID- 11917383 TI - Recruitment and retention. Slip slidin' away. PMID- 11917384 TI - Education. They've got a lot in common. PMID- 11917385 TI - Is there such a thing as work-related stress? PMID- 11917386 TI - Don't let companies milk profits from mothers. PMID- 11917387 TI - Safety first. PMID- 11917388 TI - A certain je ne sais quoi. PMID- 11917389 TI - A shock to the system. PMID- 11917390 TI - Front-runners. PMID- 11917391 TI - Nursing with dignity. Part 1: Judaism. PMID- 11917392 TI - Implementing clinical supervision. PMID- 11917393 TI - Reflecting on your expert practice. PMID- 11917394 TI - Rheumatology. 2. The diagnosis, assessment and management of complex rheumatic disease. PMID- 11917395 TI - Patient group directions. PMID- 11917396 TI - Prescribing catheters. PMID- 11917397 TI - Education and nurse prescribing. PMID- 11917398 TI - Candidiasis of the skin and acne. PMID- 11917399 TI - Do you speak nursing? PMID- 11917400 TI - Prime time for drugs. PMID- 11917401 TI - Safe home. PMID- 11917402 TI - Affairs of the heart. PMID- 11917403 TI - Let us spray. PMID- 11917404 TI - Bad old days. PMID- 11917405 TI - Patient information. 3. Healthy eating for a healthy heart. PMID- 11917406 TI - The organisation of death and dying in today's society. AB - BACKGROUND: This literature review explores service provision for dying people in the wider context of changes in the modern hospice movement, recent government reforms and, in particular, the role of nursing in influencing and improving service delivery. These government reforms are reviewed in the context of the care of the dying. The article then examines the place of death in modern society, and health professionals' views. The concept of the 'good death' is also explored and the satisfaction of patients, carers and health professionals discussed in relation to terminal care in the community. CONCLUSION: Nursing makes a valuable contribution to the care of dying patients and nurses must take the opportunities presented to them by forthcoming healthcare changes to become proactive in representing those they care for. PMID- 11917407 TI - Gaining the most from your tutor. AB - The transition to university education can prove a difficult one for nursing students, especially if they are unsure of how best to work with an academic tutor. Tutors' roles are not the same as those of school teachers or college nurse education teachers. As well as teach, they aid learning and the development of individual thought. That can pose problems for nurse students who want to get the best help from their tutor, yet are anxious that what they say or do might seem inadequate or stupid. Drawing on a five-year doctoral research programme into help-seeking among distance learners, this article describes the challenges associated with getting in touch with tutors, and recommends useful ways of building a constructive relationship. PMID- 11917408 TI - Diabetes in pregnancy. AB - A number of risk factors exist for pregnant women with diabetes and their babies. Collaborative care can ensure that women with pre-existing diabetes, and those who develop diabetes during pregnancy, receive appropriate and individualised care. PMID- 11917409 TI - Caesarean conference report: from Audit to Action. January 31, 2002. PMID- 11917410 TI - Developing professional leaders in midwifery. PMID- 11917412 TI - Herbal safety. PMID- 11917411 TI - Midwives with attitude. PMID- 11917413 TI - Partners or wreckers? PMID- 11917414 TI - Report on higher level of practice published. PMID- 11917415 TI - New framework for diabetes. PMID- 11917416 TI - Tommy's launches fight against toxoplasmosis. PMID- 11917417 TI - Employment relations advice and how to access it: helping us to help you. PMID- 11917418 TI - Antenatal care. PMID- 11917420 TI - Bournemouth 2002 conference forums: your shout. PMID- 11917419 TI - Regional Learning Networks: improving clinicians' confidence in informatics. PMID- 11917422 TI - The safety of herbal preparations. PMID- 11917424 TI - Certified registered nurse anesthetists. PMID- 11917425 TI - Proposed resolution concerning therapeutic marijuana. PMID- 11917426 TI - Our battle with infectious diseases. PMID- 11917427 TI - Allergic rhinitis. AB - Most nurses will come into contact with sufferers of allergic rhinitis, a highly prevalent disease. This Factfile examines diagnosis and management. PMID- 11917428 TI - Non-invasive positive pressure ventilation for COPD patients. AB - Patients with chronic obstructive pulmonary disease may experience acute exacerbations of their condition, leading to respiratory failure. Traditionally, patients may have been admitted to ITU for invasive mechanical ventilation but, with the introduction of non-invasive positive pressure ventilation by facial or nasal mask, more of these patients are being treated in general wards. PMID- 11917429 TI - Qualitative research on admission procedures in elderly care. AB - The admission process to hospital is vital in maintaining a patient's sense of independence and control of his or her situation, which in turn will help during the discharge procedure back to the community. This paper focuses on the qualitative research techniques used to uncover the experience of older patients and their carers during an admission to hospital. PMID- 11917430 TI - Immunotherapy. AB - Immunotherapy is effective and safe when carried out by properly trained and equipped health professionals on patients who have been carefully selected and screened. Access to these treatments is limited because, although allergy has a high profile with the public, this has yet to translate into adequate service availability. PMID- 11917431 TI - The correct use of pulse oximetry in measuring oxygen status. AB - Pulse oximetry is a valuable method of monitoring a patient's oxygenation levels and is an increasingly common test. It is important that any health-care professional conducting pulse oximetry is aware of the various factors that may influence the results obtained, ranging from incorrect use of the equipment to the effect of the patient's condition. PMID- 11917432 TI - Effective services for the care of patients with back pain. AB - For the majority of patients with back pain the traditional medical model of care is not effective. For patients without any serious underlying pathology a biopsychosocial model of care, comprising education, relaxation, exercise and analgesia, is now advocated, as practiced by multidisciplinary teams such as the Nottingham Back Team. PMID- 11917433 TI - Spinal cord stimulation for patients with chronic pain conditions. AB - Spinal cord stimulation involves the implantation of an electrical pulse generator that acts by blocking pain signals to the brain, which can be helpful in chronic pain conditions such as severe neuropathic pain. It is the role of the nurse specialist in pain management to educate and support patients undergoing this treatment. PMID- 11917435 TI - A diabetes education initiative for residential care home staff. AB - About one in eight elderly people living in residential care homes will be known to have diabetes. Lack of appropriate training for care home staff is an important barrier to improved care for this vulnerable and neglected group. A multidisciplinary, multi-agency group have begun to address this problem with a district-wide diabetes education programme for care home staff. PMID- 11917434 TI - Higher standards in diabetes care. PMID- 11917436 TI - Acupuncture and other alternative therapies in rheumatoid arthritis. AB - Increasing use of alternative therapies is being reported in many areas of health care, particularly in those specialties in which conventional medicine can offer no complete cures. This article examines what is known about the current use of alternative therapies, and acupuncture in particular, by patients with rheumatoid arthritis. PMID- 11917437 TI - Responding to the information needs of patients with cancer. AB - Cancer care has progressed from the days when doctors alone chose treatments for cancer patients and decided what information to give them. Patients are now more involved in their own care and information-giving has become a priority for nurses. However, achieving the right balance in information provision for individual patients is a challenge. PMID- 11917438 TI - [Pancreatoduodenectomy in digestive tract hemorrhage. A case report]. AB - OBJECTIVE: To describe the clinical presentation and treatment of one patient with a neuroendocrine pancreatic neoplasm and gastrointestinal hemorrhage. BACKGROUND: Neuroendocrine pancreatic tumors are relatively rare neoplasms. When the tumor does not have endocrine function, clinical manifestations are non specific and show symptoms when the tumor is quite large. Massive upper gastrointestinal hemorrhage is an infrequent clinical manifestation of these tumors. METHOD: A case of gastrointestinal hemorrhage, which required a pancreaticoduodenectomy to control bleeding of an ulcerated pancreatic head neoplasm, is presented. CONCLUSION: Pancreaticoduodenectomy is the procedure of choice to control duodenal massive bleeding secondary to an ulcerated neuroendocrine tumor of the pancreas. PMID- 11917439 TI - [CT diagnosis of appendicitis. Report of a case and review of the literature]. AB - Diagnosis of appendicitis has been historically supported in clinical findings. Imaging studies such as US and CT have been used mainly in cases where clinical diagnosis of appendicitis is uncertain. CT techniques have ranged between full abdominopelvic examinations after administration of oral and intravenous contrast media and unhenhanced scanning limited to the right bowel quadrant. We present the case report of an 18-year-old male patient with recurrent abdominal pain, with clinical suspicion of appendicitis; an abdominopelvic CT examination was performed that showed a suggestive image of appendicitis. This diagnosis was confirmed with the results of surgery and pathologic examination of the appendix. PMID- 11917440 TI - [Laparoscopic management of gastroesophageal reflux disease. Experience with 100 cases]. AB - OBJECTIVE: To evaluate the results of laparoscopic Nissen-Rossetti funduplication and to compare them with the results obtained in open surgery. DESIGN: Prospective, observational, longitudinal, pre and post-procedure. CENTERS: Beneficencia Espanola, Hospital Angeles, and Hospital Francisco Galindo Chavez, ISSSTE, in Torreon, Coahuila, Mexico. PATIENTS AND METHOD: From December 1992 to February 1999, 100 patients with surgical indications due to gastroesophageal reflux disease (GERD) prospectively underwent a laparoscopic Nissen-Rossetti procedure. A clinical and endoscopic follow up from 3 months to 9 years was performed in 87 cases. RESULTS: Symptomatic control was achieved in 98% (85/87) of the cases and remission of overall endoscopic esophagitis in 79% (69/87); excluding Barrett cases, esophagitis remission was observed in 93% (67/72) of the subjects. The following recurrences took place: two with G-II and two with G-III esophagitis, one requiring pyloroplasty due gastric stasis, and other patient with G-IV esophagitis, who has needed to continue with postoperative dilations. Of 16 cases with Barrett's esophagus, two-showed remission and one did not return control. Perioperative complications included gastric perforations (3), acute pulmonary edema during the immediate postoperative period (1), deep vein thrombosis (1), and late esophageal perforation (1). All were resolved satisfactorily. Surgical mortality was 0 in the 100 cases undergoing the procedure. Eighty-six percent of cases had a 24-h hospital stay. Early morbidity: dysphagia in 60 patients, early satiety in 91 cases, abdominal distention in 25 cases, all this symptomatology disappears during the subsequent 3 months. Persistent morbidity: flatulence in 60% of patients, difficulty for vomiting in 10% of cases. CONCLUSION: The laparoscopic procedure is as effective as the open method with the advantage of being minimally invasive. PMID- 11917441 TI - [Insertion of biliary endoprosthesis as ambulatory procedure]. AB - BACKGROUND: Because of possible complications, it has been common practice to admit to the hospital most if not all patients undergoing therapeutic ERCP. Therefore, little descriptive data exist on the safety of out-patient therapeutic ERCP for endobiliary stent placement. OBJECTIVE: To assure quality patient care and patient safety, we reviewed our experience with out-patient therapeutic ERCP for palliation of benign and malignant common bile duct obstruction by means of endobiliary stent insertion. PATIENTS-METHODS AND RESULTS: A retrospective review of all therapeutic ERCPs for palliation of benign and malignant common bile duct obstruction with endobiliary stents was performed from January 1, 1998, through December 31, 1999. One hundred forty therapeutic ERCPs were performed to place an endobiliary stent for benign and malignant common bile duct obstruction. Seventy two procedures were performed on out-patients, sixty-eight on in-patients. There was no significant difference between out-patient and in-patient groups with regard to age, gender, need for endoscopic sphincterotomy, and complication rate. In patients had one procedure-related complication; out patients had four. There was no procedure-related mortality in either group. CONCLUSIONS: Therapeutic ERCP for palliation of benign and malignant common bile duct obstruction with endobiliary stents can be safely and successfully performed on an out-patient basis for selected patients. This should result in substantial cost savings. PMID- 11917442 TI - [Transmesenteric intestinal plication for intestinal occlusion secondary to disseminated adhesions. A 12-year experience]. AB - OBJECTIVE: To report the experience with intestinal plication in patients with adhesive intestinal obstruction that was followed up to 12 years. BACKGROUND: To diminish the high recurrence rate of adhesive intestinal obstruction, there are surgical techniques of intestinal plication. In 1977 Blanco modified a pre existing transmesenteric technique that is the used in our institution. METHOD: We studied the medical records of 32 patients who underwent intestinal plication using the transmesenteric technique. The postoperative evaluation was based in recurrence and mortality. RESULTS: There were 32 patients, 56% women and 44% men, with a mean age of 50 years. All patients had a history of intraabdominal surgical procedures. The postoperative evaluation was satisfactory. The success rate was over 90%. The recurrence rate was 9.3% and we had no mortality. The mean follow-up was 3.5 years (median 3 [range 1-12] years). There were no significant differences between this technique and the Noble and Childs-Phillips plication techniques. CONCLUSIONS: This technique of intestinal plication is useful in the surgical management of patients with adhesive intestinal obstruction. PMID- 11917443 TI - [Frequency of gastric lymphoma at 6 hospitals in Mexico City]. AB - BACKGROUND: Gastric lymphoma has been traditionally considered a rare neoplasm that constitutes 1-5% of malignant gastric tumors. Two studies performed in Mexico in 1960 and 1966 found that only 1.9% and 1% of gastric neoplasms were lymphomas. Nevertheless, some studies made in the U.S. and in some European countries in recept decades have revealed an increase in the frequency of this neoplasm. A recent study made at two National Health Institutes in Mexico City (Instituto Nacional de Cancerologia and Instituto Nacional de la Nutricion) revealed a remarkable increase in the frequency of gastric lymphoma (9.3% and 10.3%, respectively) in recent years. AIM: To define whether there is an actual increase of lymphoma in our population and whether it includes other hospitals in Mexico City that provides attention to populations different from those who attend referral centers. MATERIALS AND METHODS: Six hospitals in Mexico City were selected, including two National Health Institutes (Instituto Nacional de la Nutricion and Instituto Nacional de Cancerologia), two private general hospitals used by patient with a high socioeconomic level (Hospital Espanol and Hospital Ingles), and two public general hospitals frequented by low-income patients (hospital Juarez and Hospital General de Mexico). In each case, the gastric lymphomas diagnosed in each participant hospitals in the last 5 years were registered. For comparative purpose, diagnosed cases of gastric adenocarcinoma during the same period were also registered. Other types of gastric neoplasms were excluded from the study because they formed a very heterogeneous group and represented a minimal proportion of malignant gastric tumors. Age and sex of each patient were included for all lymphomas. RESULTS: A total of 879 malignant gastric neoplasms were included in our study. The relative percentage for gastric lymphoma by institution in descendent order was Hospital Espanol 25.4%; Instituto Nacional de la Nutricion 13.7%, Hospital Ingles 11.5%, Hospital General de Mexico 8.5%, Instituto Nacional de Cancerologia 6%, and Hospital Juarez 6%. Mean general frequency taking into account the six hospitals was 9.1%. CONCLUSIONS: Frequency of gastric lymphomas in all analyzed institutions was higher than that reported in most series in the medical literature (1-5%) and that reported for the Mexican population in 1960 and 1966. The increase was most remarkable in hospitals attended by patients with high incomes (Hospital Espanol, Hospital Ingles), although the total number of neoplasms reported by these institutions was smaller than that reported by hospitals were by patients with lower incomes (Hospital Juarez, Instituto Nacional de Cancerologia). The reason for this increase is unknown, but one might speculate that some strains of Helicobacter pylori, nutritional factors, and ethnic differences could be involved. Both gastroenterologists and pathologists must recognize the increase of this neoplasm because unlike gastric adenocarcinoma, gastric lymphoma is a curable disease in a high percentage of cases. PMID- 11917444 TI - [Prevalence of Barrett's esophagus in patients with colorectal carcinoma]. AB - OBJECTIVE: To determine Barrett's esophagus (BO) prevalence in patients diagnosed with colorectal carcinoma (CC) and feasibility-associated factors in a 10 week period from March 1st to June 15, 1999. PLACE: Endoscopy Department, Oncology Hospital, Centro Medico Nacional Siglo XXI, IMSS. METHOD: A prospective, transversal, observational and comparative assay of 48 patients with histologically confirmed diagnosis of colorectal carcinoma in whom high endoscopy with biopsy harvest was performed; otherwise, colonoscopy was performed in eight patients with confirmed BO. RESULTS: Thirty men and 18 women were studied with an average age of 53 years of age. In 100%, colon cancer was determined as adenocarcinoma and location were rectal in 43.7%. Prevalence of BO was 22.9% in addition to 0.74 to 2% in the general population. This implies an odds ratio calculated at 36.43 (p = 0.00000) and 3.09 p = 0.00007), respectively, for risk to present BO concomitant with CC. There was no statistical significance in relation to age, smoking habit, and/or alcoholism. Among eight patients with BO without CCR, only one had a tubulovellous polypus (12.5%) identified. CONCLUSIONS: Although the results were not to conclusive to affirm the association of BA with CCR, the high prevalence found justifies the performance of routine endoscopy in CCR carrier patients with reflux symptoms. PMID- 11917445 TI - [Prevalence of adenomas and carcinomas of the colon. Results of the rectosigmoid exam]. AB - Colon adenomas are precursors for colon carcinoma. Their prevalence has been estimated at 20-30% in the general population and 40-60% in developed countries. They are classified as tubular, villous; or mixed, and as low- or high-risk lesions, which have any of the following characteristics: Being larger than 1 cm; having a villous component, or showing moderate to severe dysplasia. OBJECTIVE: To analyze the results of a series of colonoscopies, carried out from 1993 to 1999 in a private hospital in Mexico City, with regard to the following: Prevalence of colon adenomas and carcinoma prevalence in different age groups, distribution by gender; frequency of single vs synchronous lesions; presence or absence of distal and/or proximal lesions as markers of neoplasia and finally, prevalence of proximal lesions in the absence of distal lesions. METHODS: A retrospective analysis of a database of 701 colonoscopies, which the author carried out from January 1993 to December 1999. RESULTS: The prevalence of colon adenoma was 14.69%. The prevalence of high-risk adenomas or carcinoma was 6.7%. In 29.72% of the cases, there were concurrent lesions. These were more common in male patients. The probability of finding a proximal lesion where there was a distal adenoma was greater (17.80%) than in the absence of the latter (6.05%), but the probability of a high-risk lesion was similar whether there was a distal lesion or not (2.73 vs 2.86%). Ninety percent of high-risk proximal lesions had no distal adenoma. CONCLUSION: It was found that the prevalence of colon adenomas for the various age groups was lower than in the figures published for developed countries. Synchronic lesions were also less common. The results of this study support the use of colonoscopy as a screening test for adenomas and carcinoma of the colon, because the absence of distal lesions does not exclude the presence of high-risk neoplasia of the proximal colon. PMID- 11917447 TI - [Toxic megacolon secondary to pseudomembranous colitis]. AB - BACKGROUND: Toxic megacolon is a rare complication of pseudomembranous colitis. OBJECTIVE: The aim of this paper is to present a rare case of toxic megacolon secondary to pseudomembranous colitis. METHODS: The chart of a 75-year-old male, who developed a toxic megacolon secondary to pseudomembranous colitis no was reviewed. RESULTS: The clinical features and outcome of a 75-year-old male with pseudomembranous colitis are depicted. The main symptoms were no ever, abdominal distention, bloody and diarrhea; the man suddenly developed a toxic megacolon and taken was to surgery. Total colectomy with proximal rectal closure was performed. The patient died within 24 hours of the abdominal procedure. CONCLUSIONS: Toxic megacolon is a rare complication of pseudomembranous colitis. Its presence should be suspected when these patients develop no colonic dilatation with associated systemic toxicity. Aggressive surgical intervention is indicated. PMID- 11917446 TI - [Usefulness of gel lidocaine in esophageal manometry]. AB - BACKGROUND: Esophageal manometry (EM) is a functional study performed without sedation. The insertion of a catheter through nasal passages provokes the greatest discomfort as it is being performed. OBJECTIVE: To evaluate the usefulness of lidocaine jelly to diminish discomfort patient during EM and to ease the procedure. MATERIAL AND METHODS: Ninety patients sent to EM were randomly assigned to receive lidocaine jelly at 2% (n = 45) or chlorexidine gluconate (n = 45) as a lubricant for the catheter. A solid-state catheter was employed via the pull-through technique under topic anesthesia of the hypopharynx in all cases. Patients quantified nasal pain and nausea produced by analog visual scale (AVS, 0-10 cm). The physician quantified difficulty of insertion using the same method. An independent observer kept time records from the moment of insertion of the probe through the nasal passages until the location of all sensors in the stomach (time of insertion). RESULTS: No significant differences were found between the two groups in their evaluation of nasal pain (2.8 cm lidocaine group vs. 2.6 cm chlorhexidine group), and there was no difference found in the intensity of nausea (3.3 cm vs. 3.1, respectively). Ease of the procedure was similar (2.2 cm lidocaine group vs. 1.8 cm chlorhexidine group), as well as insertion time (120 vs. 111 sec, respectively). CONCLUSIONS: Lidocaine jelly at 2% neither diminishes the discomfort provoked by the insertion of an EM catheter, nor does it make the procedure any easier. PMID- 11917448 TI - [Primary melanoma of the esophagus. Unusual case with Barrett's esophagus]. AB - OBJECTIVE: To present a rare entity of difficult preoperative diagnosis. BACKGROUND: Primary malignant melanoma of the esophagus represents 0.1% of all malignant tumors in this organ. It has been described in association with esophageal melanocytosis and melanosis with a clinic behavior similar to other neoplasm in the esophagus, but is more aggressive and fatal. By endoscopy, it usually appears as a vegetant, non-obstructive lesion, with dark pigmentation in the melanotic variant, the sole evidence that suggests the diagnosis prior to microscopic examination. Clinical exclusion of metastasis melanoma as a possibility is obligatory. METHODS: We reviewed clinical, imagenologic, endoscopic, and anatomopathological aspects of an ulcerovegetant neoplasm on the esophago-gastric junction, surgically resected, in a 65 years-old man with progressive dysphagia, who was without tumoral relapse during 18 months after surgery. RESULTS: The tumoral mass was an anaplastic cell tumor with very scant melanic pigmentation and diffuses stain for HMB-45 antigen and S100 protein, considering it to be a primary malignant melanoma. The neighboring mucous membrane showed a typical Barrett's esophagus and melanocytic hyperplasia. CONCLUSIONS: 1) Endoscopy may suggest the possibility of esophagus melanoma when there is a vegetate, non-obstructive, pigmented tumor. 2) Immunohistochemistry confirms the entity if HMB-45 antigen and S100 protein are detected in the tumoral cells, while the negative results for keratin and leukocytic common antigen (LCA) eliminate carcinoma and lymphoma as possible diagnoses. 3) In our case, the presence of Barrett's esophagus is exceptional, because it has not been described previously in the literature reviewed. PMID- 11917449 TI - [Melanoma of the bile ducts. Report of a case and review of the literature]. AB - Primary malignant melanomas of the bile ducts are extremely rare, with only seven cases previously reported in the literature published in English. This report concerns a 67-year-old woman with a primary melanoma at the confluence of the hepatic ducts. She underwent resection and hepaticojejunostomy. Seventeen years later, she remains alive with no evidence of recurrence. A thorough review of the literature was done. Surgical resection can offer hope for long-term survival. PMID- 11917450 TI - [Obstructive esophageal membrane in recessive dystrophic epidermolysis bullosa]. AB - BACKGROUND: Dysphagia is a rare complication of esophageal webs. OBJECTIVE: To describe a patient with dystrophic epidermolysis bullosa who developed dysphagia produced by an esophageal web that simulated carcinoma. CASE REPORT: A 26 year old male with dystrophic epidermolysis bullosa developed severe dysphagia and subsequently aphagia secondary to an upper third esophageal obstruction. A (CT) scan showed a circumferential esophageal wall thickening of the affected area. The patient was submitted to surgical neck exploration with endoscopic assistance to identify the obstruction and a esophagotomy was carried on detecting a complete esophageal web that was excised. CONCLUSIONS: Esophageal web must be considered a cause of esophageal obstruction in patients with dystrophic epidermolysis bullosa. PMID- 11917451 TI - [Transjugular intrahepatic portosystemic shunt (TIPS). Thirteen-year experience]. AB - The transjugular intrahepatic portosystemic shunt (TIPS) is a non-selective portosystemic shunt that is created by using percutaneous endovascular techniques. TIPS has been employed for the last 13 years mainly for the management of hemorrhagic portal hypertension. The present review addresses important concepts including the accepted indications, contraindications, technical results, clinical results (bleeding control, management of refractory ascites, and other complications of portal hypertension), most important complications, and a brief overview of the most recent research in TIPS. PMID- 11917452 TI - [Colonic cancer in Mexico?]. PMID- 11917453 TI - Cutaneous manifestations of endocrine disorders. AB - Endocrine disorders such as diabetes or Cushing's syndrome often have cutaneous findings, including changes to the skin, hair, and nails. In this review, the major causes, clinical manifestations, laboratory workup, and treatment of the most important endocrine diseases are discussed. PMID- 11917454 TI - Sezary syndrome: a summary. AB - Sezary syndrome is the leukemic form of primary cutaneous T-cell lymphoma. It is an aggressive disease, with the lowest reported median survival of all cutaneous lymphomas. Patients with Sezary syndrome live with the awareness that they are suffering from an incurable disease. Having to cope daily with extensive skin care regimens, these patients can benefit tremendously from the expertise of dermatology nurses, who can teach them skin selfcare and who are aware of the psychologic impact of this disease. The symptoms, treatments, and emotional distress related to Sezary syndrome are summarized. PMID- 11917455 TI - Cutaneous malignant melanoma. AB - Melanoma is the deadliest form of skin cancer; 47,700 persons were expected to be diagnosed and 9,600 were expected to die from melanoma in 2000 (American Cancer Society, 2000). It is important for dermatology nurses to understand the epidemiology, risk factors, clinical presentation, diagnosis, treatment, prognosis, and prevention of cutaneous malignant melanoma. PMID- 11917457 TI - Wound assessment and evaluation. AB - The "Wound Assessment and Evaluation" series includes a short case presentation and differential diagnosis. It is followed by a discussion of the disease or condition and the rationale used in each step of the assessment. PMID- 11917458 TI - Pruritic erythematous, indurated plaques on the feet. AB - The "Clinical Snapshot" series provides a concise examination of a clinical presentation including history, treatment, patient education, and nursing measures. Using the format here, you are invited to submit your clinical snapshot to Dermatology Nursing. PMID- 11917456 TI - The WOC nurse: economic, quality of life, and legal benefits. AB - Patients who require treatment for complications stemming from skin and wound care conditions represent a significant and costly clinical problem in the health care arena. An innovative approach used to manage these patients is utilization and employment of the specialty trained wound, ostomy, and continence (WOC) nurse. A review of the literature supports the role of the WOC nurse in providing quality care, minimizing the expenditure of health care dollars, and overseeing the use of pressure ulcer and wound care guidelines so that litigious events may be avoided. PMID- 11917459 TI - What's your assessment? Dermatitis medicamentosa. AB - The "What's Your Assessment?" series includes a short case presentation and differential diagnosis. It is followed by a discussion of the disease or condition and the rationale used in each step of the assessment. PMID- 11917460 TI - Psoriasis update. PMID- 11917461 TI - Acne news. PMID- 11917462 TI - Skin cancer update. PMID- 11917463 TI - [History of liver function tests]. PMID- 11917465 TI - [Endoscopy]. PMID- 11917464 TI - [Imaging diagnosis]. PMID- 11917466 TI - [Laparoscopic surgery]. PMID- 11917467 TI - [Hepatic biopsy]. PMID- 11917468 TI - [Digestive system hormones and host control system]. PMID- 11917469 TI - [Helicobacter pylori]. PMID- 11917470 TI - [Bile acid and gallstone resolving agents]. PMID- 11917471 TI - [Hepatitis viruses]. PMID- 11917472 TI - [Autoimmune hepatitis]. PMID- 11917473 TI - [Transplantation of organs of the digestive system]. PMID- 11917474 TI - [Endoscopic diagnosis of cancer of the digestive system]. PMID- 11917475 TI - [Endoscopic treatment of cancers of the digestive system]. PMID- 11917476 TI - [Transcutaneous transhepatic cholangiography and bile drainage]. PMID- 11917477 TI - [Treatment of hepatic cancer by transcutaneous ethanol injection]. PMID- 11917478 TI - [Esophageal cancer]. PMID- 11917479 TI - [Peptic ulcer]. PMID- 11917480 TI - [Stomach cancer]. PMID- 11917482 TI - [Inflammatory bowel diseases]. PMID- 11917481 TI - [Colonic cancer]. PMID- 11917483 TI - [History of cholelithiasis]. PMID- 11917484 TI - [Biliary tract cancer]. PMID- 11917486 TI - [Hepatocellular carcinoma]. PMID- 11917485 TI - [Viral hepatitis and liver cirrhosis]. PMID- 11917487 TI - [Pancreatic cancer]. PMID- 11917488 TI - [Pancreatitis]. PMID- 11917489 TI - [Activities in the study of the digestive system: reflection on the 20th century and an overview of the coming century. A discussion]. PMID- 11917491 TI - [Therapeutic guideline reviews. Stomach cancer--the Japanese Society of Stomach Cancer]. PMID- 11917490 TI - [Guideline reviews: Helicobacter pylori infection]. PMID- 11917492 TI - [Guideline for therapy and diagnosis of gastric ulcer: an interim report by a research committee of the Department of Health, Labor, and Welfare]. PMID- 11917493 TI - [Guideline reviews: inflammatory bowel disease]. PMID- 11917494 TI - [Guideline reviews: Chronic pancreatitis]. PMID- 11917495 TI - [Frontotemporal dementia--the usefulness of surface anatomy scanning for evaluation of brain atrophy with MRI]. PMID- 11917496 TI - [Idiopathic thrombocytopenic purpura treated with splenic irradiation]. PMID- 11917497 TI - [A catecholamine-secreting glomus jugular tumor]. PMID- 11917498 TI - [Acute autonomic and sensory neuropathy successfully treated with intravenous immunoglobulin therapy]. PMID- 11917499 TI - [Primary pericardial B-cell lymphoma with complete atrioventricular block]. PMID- 11917500 TI - [Three cases of Takotsubo cardiomyopathy--reversible left ventricular dysfunction]. PMID- 11917501 TI - [A longer prognosis of B-CLL with mutation of IgVH gene]. PMID- 11917502 TI - [Two cases of rickettsiosis--the clinical characteristics of Japanese spotted fever and tsutsugamushi disease]. PMID- 11917503 TI - [Multiple myeloma light chain deposition with specific pathological change]. PMID- 11917504 TI - [POEMS syndrome with increased vascular endothelial growth factor (VEGF)]. PMID- 11917505 TI - Identification of rosmarinic acid as a novel antidepressive substance in the leaves of Perilla frutescens Britton var. acuta Kudo (Perillae Herba). AB - The leaves of Perilla frutescens Britton var. acuta Kudo (Perillae Herba) are found in some traditional oriental herbal medicines that are primarily used to treat affective disorders such as depression and anxiety. The aim of the present study was to identify the bioactive component in Perillae Herba that possesses antidepressive activity. The effects of a water extract of Perillae Herba and six fractions therefrom were evaluated in mice by use of the forced-swimming test. An oral administration of a water extract of Perillae Herba significantly reduced the duration of immobility. Moreover, 50% methanol extract of the water extract of Perillae Herba and its 30% methanol extract also reduced the duration of immobility. Three-dimensional high-performance liquid chromatography and FAB-MS and NMR spectral analysis clearly showed that the extracts with anti-immobility effects contained abundant rosmarinic acid. The oral and intraperitoneal administration of rosmarinic acid significantly reduced the duration of immobility. In contrast, a water extract from another species of Perillae Herba, which contains only low levels of rosmarinic acid, had no anti-immobility effect. These results suggest that rosmarinic acid may be the main component involved in the antidepressive effect of Perillae Herba in the forced-swimming test. Thus it may be a novel antidepressive substance. PMID- 11917506 TI - [Modulation of neuronal activities in the central nervous system via sigma receptors]. AB - Sigma receptors have recently been the target of drug development related to psychiatric disorders, including schizophrenia and depression, as well as cognitive enhancers. This paper focused on the sigma-receptor-mediated modulation of neuronal activity, especially the effects on aminergic neuron and hippocampal neuron activity. Dopaminergic neuron activities in the substantia nigra and ventral tegmental area (VTA) are variously modified by the systemic administration of sigma ligands. When applied with microiontophoresis, they are reported to increase dopaminergic neuron activity in the VTA. This activity may be involved in the psychotropic or antipsychotic effects of these ligands. Moreover, serotonergic neurons in the raphe nucleus and noradrenergic neurons in the locus coeruleus were activated by sigma ligands. These effects are probably related to the antidepressant activity of sigma receptor ligands. In the hippocampus, sigma ligands suppressed CA1 neuronal activity in vitro. The effects were suggested to be due to an increase in the threshold of action potential and decreased synaptic transmission efficacy. NMDA receptor function was modified in biphasic fashion related to doses of sigma ligands, that is, a lower dose facilitated the NMDA receptor functions, and a higher dose inhibited them. These effects on the hippocampal neurons may contribute to their neuroprotective and antiamnesic actions. Further studies are needed to elucidate the relation between the physiological function of sigma receptor and psychiatric diseases by the use of sigma receptor ligands and molecular techniques. PMID- 11917510 TI - Limited powers. PMID- 11917507 TI - [Glutamate hypothesis of schizophrenia and targets for new antipsychotic drugs]. AB - N-methyl-D-aspartate (NMDA) receptor antagonists such as phencyclidine (PCP) and ketamine have been known to cause schizophrenia-like psychosis (positive symptoms, negative symptoms, cognitive dysfunction) in humans. A dysfunction of glutamatergic neurotransmission may play an important role in the pathophysiology of schizophrenia. In this review, the glutamate hypothesis of schizophrenia, especially the mechanism of neurotoxicity of NMDA receptor antagonist in the posterior cingulate cortex and retrosplenial cortex of the brain, is summarized. Furthermore, the roles of the posterior cingulate cortex and the retrosplenial cortex in the pathophysiology of schizophrenia and Alzheimer's disease are also discussed. Moreover, the glycine site of the NMDA receptor, metabotropic glutamate receptor, AMPA receptor, and antioxidant glutathione as novel potential targets for the treatment of schizophrenia are discussed. PMID- 11917511 TI - Home comforts. PMID- 11917512 TI - Feeling the freeze. PMID- 11917513 TI - Non-pharmacological treatment of breathlessness. AB - BACKGROUND: Breathlessness is a common and distressing symptom in cancer patients, and medical interventions to relieve it can often cause some distress. Patients can be taught self-help measures, but they might not remember them later and nursing staff might not be aware of these measures and so cannot reinforce them. CONCLUSION: A poster has been designed to remind patients of non pharmacological methods of relieving dyspnoea and to educate staff and agency nurses on postural changes and breathing retraining. It is hoped that this will result in more holistic care for cancer patients with shortness of breath. PMID- 11917514 TI - Clinical effectiveness and evidence-based practice. AB - An essential component of clinical governance is clinical effectiveness, which is achieved through evidence-based practice. This has enormous implications for the way nurses work, and the qualitative approach often taken by nurse researchers can be difficult to incorporate into the new systems. Clinical governance should have benefits for patients and NHS staff alike. If the goals are to be achieved, a multidisciplinary approach will be needed, with greater encouragement being given to creative research methods. In addition, nurses will have to become more powerful inside healthcare organisations. PMID- 11917515 TI - Pulmonary hypertension. AB - Pulmonary hypertension (PH) is a rare blood vessel disorder of the lung, in which the pressure in the pulmonary artery rises above normal levels and can become life-threatening. The authors discuss the disease process, symptoms, investigations and patient management. PMID- 11917516 TI - Cultural shift. PMID- 11917517 TI - Transepithelial measurements of bicarbonate secretion in Calu-3 cells. PMID- 11917518 TI - Transepithelial impedance analysis of chloride secretion. PMID- 11917519 TI - Studies of the molecular basis for cystic fibrosis using purified reconstituted CFTR protein. PMID- 11917520 TI - Probing CFTR channel structure and function using the substituted-cysteine accessibility method. PMID- 11917521 TI - Methods for the study of intermolecular and intramolecular interactions regulating CFTR function. PMID- 11917522 TI - Fluorescent indicator methods to assay functional CFTR expression in cells. PMID- 11917523 TI - Immunolocalization of CFTR in intact tissue and cultured cells. PMID- 11917524 TI - cDNA microarrays for pharmacogenomic analysis of cystic fibrosis. PMID- 11917525 TI - Analysis of CFTR trafficking and polarization using green fluorescent protein and confocal microscopy. PMID- 11917526 TI - CFTR folding and maturation in cells. PMID- 11917527 TI - Isolation of CFTR. Chaperone complexes by co-immunoprecipitation. PMID- 11917529 TI - Manipulating the folding pathway of delta F508 CFTR using chemical chaperones. PMID- 11917528 TI - CFTR expression and ER-associated degradation in yeast. PMID- 11917530 TI - CFTR degradation and aggregation. PMID- 11917531 TI - In vitro reconstitution of CFTR biogenesis and degradation. PMID- 11917532 TI - CFTR mutation detection by multiplex heteroduplex (mHET) analysis on MDE gel. PMID- 11917533 TI - Natural animal models of human genetic diseases. PMID- 11917534 TI - In vitro CFTR folding assays. PMID- 11917535 TI - Analysis of CFTR endocytosis by cell surface biotinylation. PMID- 11917536 TI - CFTR regulation of ENaC. PMID- 11917537 TI - Yeast two-hybrid identification and analysis of protein interactions with CFTR. PMID- 11917538 TI - Biochemical assays for studying indirect interactions between CFTR and the cytoskeleton. PMID- 11917539 TI - CFTR-associated ATP transport and release. PMID- 11917540 TI - Inflammatory mediators in CF patients. PMID- 11917541 TI - Bacterial colonization and infection in the CF lung. PMID- 11917542 TI - Antimicrobial peptides and proteins in the CF airway. PMID- 11917543 TI - Bacterial-epithelial interactions. PMID- 11917544 TI - Thin-film measurements of airway surface liquid volume/composition and mucus transport rates in vitro. PMID- 11917545 TI - Electrophysiological approach to studying CFTR. PMID- 11917546 TI - Murine models of CF airway infection and inflammation. PMID- 11917547 TI - Analysis of lipid abnormalities in CF mice. PMID- 11917548 TI - Bioelectric measurement of CFTR function in mice. PMID- 11917549 TI - Xenograft model of the CF airway. PMID- 11917550 TI - Development of conditionally immortalized epithelial cell lines from CF and non CF mice. PMID- 11917551 TI - Technical approaches to analyze the in vivo ion composition of airway surface liquid. PMID- 11917552 TI - Design of gene therapy clinical trials in CF patients. PMID- 11917553 TI - Formulation of synthetic vectors for cystic fibrosis gene therapy. PMID- 11917554 TI - Adeno-associated viral vectors for CF gene therapy. PMID- 11917555 TI - Quantitative analysis of ATP-dependent gating of CFTR. PMID- 11917556 TI - CFTR regulation by phosphorylation. PMID- 11917557 TI - [Treatment of alcoholic hepatitis]. PMID- 11917558 TI - [Hepatorenal syndrome]. PMID- 11917559 TI - [New treatments of variceal hemorrhage]. PMID- 11917560 TI - [Treatment of hepatocarcinoma]. PMID- 11917561 TI - [Liver-assist non-biological artificial systems]. PMID- 11917562 TI - [Living donor liver transplantation in adults]. PMID- 11917563 TI - [Treatment of hepatic encephalopathy]. PMID- 11917564 TI - Code breaker. PMID- 11917565 TI - Changes provoke debate. The new rules make it clear that nurses are still nurses even after they have finished their shift. PMID- 11917566 TI - Complete turnaround. Tears, long trolley waits and a hostile local press characterised A&E in Bath. PMID- 11917568 TI - Caring for people with depression. AB - BACKGROUND: Mental health care has recently become a government priority, and depression is a very common mental health problem. Many people feel that the mode of treatment used is less important than the effective delivery of care and intensity of follow up. The authors of this study devised a questionnaire to elicit the views of a small sample of mental health professionals and service users on the quality of current services. CONCLUSION: Most respondents felt that there is room for improvement in mental health services. Perceptions of the needs of people with depression focused on care, rather than on specific interventions. The voluntary sector was rated highly in caring for people with depression. Greater awareness among professionals of self-help organisations was seen as beneficial. PMID- 11917569 TI - Legal and professional aspects of intravenous therapy. AB - The nurse's role in intravenous therapy can result in professional satisfaction, but also increased professional risk and liability. The author argues that nurses must have a clear understanding of the legal and professional aspects of intravenous therapy to maintain their professional integrity. PMID- 11917570 TI - Central venous catheters and central venous pressure. AB - Central venous catheters are increasingly being used in a variety of clinical areas outside critical care. Philip Woodrow examines the indications, measuring techniques and complications associated with central venous pressure monitoring. PMID- 11917572 TI - Cervicovaginal smear abnormalities in sexually active adolescents. Implications for management. AB - OBJECTIVE: To assess the prevalence and spectrum of Pap smear (PS) abnormalities in sexually active adolescents in comparison to adult women in order to determine whether management of adolescents should differ from that of adults. STUDY DESIGN: Five hundred twenty-four adolescents who had an initial PS at our institution from January to September 1997 were followed for 36 months with repeat PS and/or cervical biopsy. Initial PS results were compared with those of adult women. The chi 2 test was used to calculate the statistical significance of differences between the two groups. The qualified atypical squamous cells of undetermined significance (ASCUS) cytologic diagnosis in adolescents was correlated with follow-up data. RESULTS: The overall prevalence of squamous intraepithelial lesions (SILs) in adolescents was 29% as compared to 23% in adults. Almost all initial squamous lesions were ASCUS and low grade squamous intraepithelial lesion (LSIL); only one case of high grade squamous intraepithelial lesion (HSIL) was detected. On follow-up 18% and 2.4% of adolescents developed LSIL and HSIL, with a LSIL/HSIL ratio of 8/1 as compared to 5/1 in adults. The average time from initial PS to detection of HSIL was 20 months. All patients with HSIL except one had had one or more previous abnormal PSs. The positive predictive values (PPVs) for subsequent dysplasia for ASCUS favor reactive (ASCUS.R), ASCUS not otherwise specified (ASCUS.NOS) and ASCUS favor dysplasia (ASCUS.D) in adolescents were .13, .17 and .31, respectively. ASCUS.NOS (P = .01) and ASCUS.D (P = .007) were strong indicators of dysplasia as compared to ASCUS.R. CONCLUSION: PS abnormalities are more common in sexually active adolescents, with a significantly higher prevalence of LSIL over HSIL as compared to adult women. Given the natural history of HPV infection, we recommend follow-up with cytology rather than colposcopy/biopsy for adolescents with ASCUS and LSIL PSs. Qualification of ASCUS is useful in determining which adolescents are at the highest risk of cervical dysplasia. PMID- 11917573 TI - Effectiveness of mass screening for endometrial cancer. AB - OBJECTIVE: To investigate the effectiveness of mass screening for endometrial cancer using Endocyte (Laboratoire CCD, Paris, France) endometrial smears. STUDY DESIGN: The study subjects were consecutive patients with documented endometrial cancer diagnosed between January 1, 1989, and December 31, 1997, at 22 hospitals in Japan. One hundred twenty-six cases were detected by mass screening and 1,069 diagnosed in outpatient clinics. We compared the stage of cancer at diagnosis and survival rate of patients in the two groups. RESULTS: Early stage was significantly more frequent in the screening group (P < .001); stage I comprised 88.1% of the screening group as compared with 65.3% of the outpatient group. Well differentiated adenocarcinoma was significantly more frequent in the screening group (P < .01); grade 1 constituted 74.7% of the screening group as compared with 61.0% of the outpatient group. The five-year survival rate was significantly higher in the screening group than in the outpatient group (94.0% vs. 84.3%, P = .041). The crude hazard ratio (HR) of dying of endometrial cancer for the screening group as compared to the outpatient group was .47 (95% CI .22-.99, P = .048). HR became .96 (95% CI .45-2.08, P = .925) after adjustment for age, study area and cancer stage. CONCLUSION: The results suggest that an endometrial cancer screening program would lead to early detection and improved survival among women with endometrial cancer. PMID- 11917574 TI - Diagnosing endometrial carcinoma with cervical involvement by cervical cytology. AB - OBJECTIVE: To assess the relationship of a cervical cytologic diagnosis based on number, size and degeneration of malignant clusters and necrotic background to cervical involvement of endometrial carcinoma. STUDY DESIGN: Cervical smears of 53 women with endometrial carcinoma were evaluated for cervical involvement. The cytologic diagnosis was compared with actual involvement, and accuracy was calculated. Retrospectively, cytologic features, including number, size and degeneration of malignant clusters and necrotic background, were analyzed in involved and noninvolved cases. RESULTS: Cervical involvement was confirmed in 15 patients (28.3%). The number and size of malignant clusters in the involved cases were significantly larger than those in the noninvolved cases (P < .001 and < .01, respectively). The proportion of degenerated malignant cells and necrotic background in involved cases were significantly higher than those in noninvolved cases (P < .05). Cytologic diagnosis had a sensitivity and specificity of 62.5% and 86.8%, respectively. CONCLUSION: Cervical smears of involved cases revealed a large number and large size of malignant clusters. These findings support cytologic diagnosis based on number, size and degeneration of malignant cells and necrotic background. Cervical cytology is useful to exclude cervical involvement because of its high specificity and can help detect cervical involvement because of its moderately high sensitivity. PMID- 11917575 TI - Use of automated primary screening on liquid-based, thin-layer preparations. AB - OBJECTIVE: To evaluate the accuracy of the AutoPap System (TriPath Imaging, Inc., Burlington, North Carolina, U.S.A.) (TriPath) in screening AutoCyte PREP liquid based, thin-layer preparations by comparing the final cytologic diagnoses with instrument slide classification results. STUDY DESIGN: A total of 9,665 AutoCyte PREP thin-layer slides were first independently screened to establish a final cytologic diagnosis (reference diagnosis). The slides were then processed on the AutoPap System. Each slide successfully processed was reported into result categories. The generated report gave a ranking score for each slide designated for "review." Slides designated "no further review" (NFR) were also listed in the report. The reported results were then compared to the reference cytologic diagnoses. RESULTS: Of 9,665 slides initially submitted to the AutoPap, 8,688 (90.8%) were qualified for scanning, and 884 (9.2%) were definitely classified as process review or rerun and excluded from the study. Of high grade squamous intraepithelial lesions and greater (HSIL+), 85.2% were ranked in the first rank, 12.7% in the second, one (2.1%) in the third, none in the fourth and fifth and none in the NFR category. Of low grade squamous intraepithelial lesions, 47.4% were ranked in the first rank, 20.8% in the second, 10.6% in the third, 10.1% in the fourth, 5.3% in the fifth and 5.8% in NFR. Of atypical squamous cells of undetermined significance and atypical glandular cells of undetermined significance, 53.6% were ranked in the first rank, 22.5% in the second, 12.4% in the third, 5.4% in the fourth, 3.8% in the fifth and 2.3% in NFR. Considering a cutoff value at < or = 3rd rank, 84% of cervical abnormalities (RR 6.52, 95% CI 4.96-8.66) and 100% of HSIL+ were identified. CONCLUSION: The AutoPap demonstrates a high capability for detecting cervical abnormalities on AutoCyte PREP thin-layer slides. HSIL+ was associated with the highest instrument scores. PMID- 11917576 TI - Validation of AutoPap primary screening system sensitivity and high-risk performance. AB - OBJECTIVE: To confirm or deny the reported sensitivity of the AutoPap Primary Screening System (AutoPap) (TriPath Imaging Inc., Burlington, North Carolina, U.S.A) in a moderate-sized laboratory and to determine performance characteristics for the "clinically high risk" (CHR) patient population. STUDY DESIGN: Archives were searched for low and high grade squamous intraepithelial lesion (LSIL, HSIL), adenocarcinoma in situ (AIS) and cancer (Ca) with follow-up biopsies demonstrating a lesion of at least the reported Pap smear's severity. Smears fulfilling these criteria and a matched normal, control slide from the same day of preparation were subjected to evaluation on the AutoPap. Two hundred eighty-three smears from 254 patients were enrolled in the study, including 80 LSIL, 178 HSIL, 5 AIS and 20 Ca. Specific criteria established CHR status. Fisher's exact test was applied to determine AutoPap performance differences for non-CHR and CHR populations. RESULTS: AutoPap successfully classified as "Review" all cases as follows: 91.2% LSIL (73/80), 96.6% HSIL (172/178), 100% AIS (5/5), and 100% Ca (20/20). Fisher's exact P values, pLSIL = 1.00 and pHSIL+ = .411, confirmed statistically equivalent performance. CONCLUSION: The results confirmed the sensitivity data reported in the Food and Drug Administration-approved labeling of the AutoPap and indicated no statistically significant differences in performance characteristics in a CHR population when compared to patients without CHR status for all grades of abnormality examined. PMID- 11917578 TI - AgNORs as an early marker of sensitivity to radiotherapy in gynecologic cancer. AB - OBJECTIVE: To evaluate the changes induced in silver-stained nucleolar organizer regions (AgNORs) by the first fraction of a radiotherapy protocol for gynecologic cancer on exfoliated cytologic samples to predict the therapeutic success of the full protocol. STUDY DESIGN: Thirteen gynecologic cancer patients who were scheduled for radiotherapy were included in the study. Cell smears were taken from the affected area before and after the first fraction of a radiotherapy protocol and silver stained for AgNORs. AgNORs per nucleus were counted under a light microscope. Local disease control by the full radiotherapy protocol was assessed at one year by the Papanicolaou technique. RESULTS: Local success of radiotherapy was greater for lesions with higher pretreatment AgNOR counts and for lesions that underwent a greater percentage reduction in AgNOR counts after the first fraction. We correlated local success of the full radiotherapy protocol with a predictive index based on AgNOR counts obtained before and after the first fraction. CONCLUSION: A predictive index based on AgNOR counts can predict, as early as after the first fraction, the local control of disease by a full radiotherapy protocol. Knowledge of the probability of success long before the protocol is completed would allow reevaluation of therapeutic options. PMID- 11917579 TI - Adenomyoepithelioma of the breast. A review of three cases with reappraisal of the fine needle aspiration biopsy findings. AB - OBJECTIVE: To describe the fine needle aspiration cytology findings of adenomyoepithelioma of the breast, with histologic correlation. STUDY DESIGN: The author reviewed the cytologic findings of three cases of adenomyoepithelioma of the breast in the files of Pamela Youde Nethersole Eastern Hospital in a three year period, 1998-2000. The diagnosis was confirmed by histologic examination, immunohistochemical study and electron microscopy. RESULTS: All cases showed similar cytologic findings. The direct smears and cytospin preparations contained an obvious biphasic cell population. There were clusters and sheets of benign apocrine cells admixed with clumps of bland-looking oval to spindle cells. The apocrine cells contained larger, round nuclei; prominent solitary nucleoli; and ample eosinophilic, granular cytoplasm. In contrast, the spindle cells had oval nuclei, fine chromatin, inconspicuous nucleoli and scanty amphophilic cytoplasm. A small number of named nuclei and foamy macrophages was noted in the background. The characteristic stromal elements seen in fibroepithelial tumor of the breast were not found. The myoepithelial nature of the spindle cells was confirmed by immunohistochemical and ultrastructural studies. Subsequent simple mastectomy and wide local excision were performed on cases 1 and 2, respectively, revealing focal carcinomatous transformation in the adenomyoepitheliomas. The carcinoma cells, however, were not sampled in the initial cytologic specimens. CONCLUSION: The cytologic features of adenomyoepithelioma, though variable and subtle, are characteristic enough for diagnosis. Recognition of the peculiar combination of benign apocrine cells and clumps of nondescript spindle cells should alert the cytologist to this rare but distinct entity, which carries a propensity for malignant transformation. PMID- 11917577 TI - Performance of monolayered cervical smears in a gynecology outpatient setting in Kuwait. AB - OBJECTIVE: To compare ThinPrep (TP) Papanicolaou smears (Cytyc Corp., Box borough, Massachusetts, U.S.A.) with matching conventional Papanicolaou (CP) smears for specimen adequacy, cytologic quality, diagnostic accuracy and screening time. STUDY DESIGN: In this prospective study of 1,024 women a split sample, matched-pair design in favor of CP slides based on single-blind criteria was followed with a smear on a glass slide for CP and the remaining material collected in Preserv-Cyt solution (Cytyc) for a TP smear. A Cytobrush (Medscand, Hollywood, Florida, U.S.A.) was used for smear preparation for CP. TP smears were processed in ThinPrep 2000 (Cytyc). Smears were stained with Papanicolaou stain and were interpreted according to the Bethesda system. RESULTS: The number of satisfactory but limited (SBL) cases with TP were 77 (7.5%) as compared to 127 (12.4%) with the CP method. This reduction in SBL smears with the TP method and consequent increase in satisfactory smears were highly significant (P < .001) by McNemar's test. As regards unsatisfactory smears in discordant pairs, although the number of unsatisfactory smears was higher with TP (41 cases) as against CP (27 cases), the difference was not statistically significant (P < .05). The split sample method showed a high correlation between the CP and TP diagnoses. TP smears had a significant advantage over CP smears in the reduction in the number of ASCUS and AGUS cases (14 vs. 29) (P < .05) and increased the pickup rate of LSIL, 6 vs. 1. Time taken to screen the TP smears was half that of CP smears. No cases of LSIL or HSIL were missed on TP smears. CONCLUSION: The liquid-based processor significantly improved the adequacy and quality of smears, resulting in fewer recall cases for SBL smears, leading to more definitive diagnoses in atypical cases, increasing the pickup rate of LSILs and reducing the screening time. A machine handling multiple specimens automatically would decrease cost and be an asset to a cytopathology laboratory. PMID- 11917580 TI - Fine needle aspiration cytology of ductal breast carcinoma with neuroendocrine differentiation. Review of eight cases with histologic correlation. AB - OBJECTIVE: To describe the fine needle aspiration cytology findings of ductal breast carcinoma with neuroendocrine differentiation and correlate them with the histologic appearance. STUDY DESIGN: We reviewed the cytologic features of eight cases of ductal carcinoma with neuroendocrine differentiation in the files of Pamela Youde Nethersole Eastern Hospital during the three-year period 1998-2000. Immunohistochemical study for neuroendocrine markers was performed, with ultrastructural correlation. RESULTS: All cases showed similar cytologic features. The smears were of moderate to high cellularity with predominantly dispersed or loosely cohesive tumor cells. The carcinoma cells were mostly of low cytologic grade. They possessed round and relatively uniform, eccentric nuclei; finely stippled chromatin; sometimes small, distinct nucleoli; and discrete cell borders. Abundant eosinophilic and focally granular cytoplasm was a common finding. In some of the cases there was accentuation of staining in the paranuclear region; it correlated with aggregates of dense core neurosecretory granules seen ultrastructurally. Mucoid substance was seen in the background in some of the aspirates. Histologic examination of the tumors showed invasive ductal carcinoma with an organoid growth pattern and sometimes mucinous component. The neuroendocrine differentiation was confirmed immunohistochemically. CONCLUSION: Although this subtype of ductal carcinoma probably carries no significant prognostic value per se, it has distinct cytologic features, rendering preoperative diagnosis possible. Recognition of this entity is important in order to avoid the misdiagnosis of neuroendocrine tumor metastatic to the breast. PMID- 11917581 TI - Phyllodes tumor of the breast. A cytomorphologic approach based on evaluation of epithelial cluster architecture. AB - OBJECTIVE: To develop more useful criteria for differentiating benign phyllodes tumor (BPT) from fibroadenoma (FA) of the breast on fine needle aspiration cytology (FNAC), with a focus on architectural features of epithelial clusters. STUDY DESIGN: The smears of 18 cases obtained by preoperative FNAC, each with tissue-proven BPT, were compared to those of 25 cases of FA. Several features of epithelial clusters in both groups were studied, with histologic correlation. RESULTS: BPT exhibited large epithelial clusters longer than 1 mm, with a wavy or folded shape, that could be distinguished from the small or medium-sized clusters with tubular, blunt-branching or monolayered contours in FA. This appearance of BPT was equivalent to the characteristic phyllodes pattern of its histology. CONCLUSION: The size and shape of epithelial clusters are important diagnostic clues in addition to several findings that have been described as differentiating BPT from FA on FNAC. PMID- 11917582 TI - Correlation of immunocytochemical and immunohistochemical determination of estrogen and progesterone receptors in breast cancer. AB - OBJECTIVE: To evaluate prospectively the degree of correlation between immunocytochemical (ICC) and immunohistochemical (IHC) determination of estrogen (ER) and progesterone receptors (PR) in breast cancer. STUDY DESIGN: Fine needle aspiration cytology (FNAC) of 101 primary breast cancers were immunostained for ER and PR. They were compared with similar determinations in formalin-fixed paraffin sections of biopsies from the same patients. In cases of discrepancy, the histologic result was considered the gold standard. RESULTS: For ER a cytohistologic correlation of 94%, with a sensitivity of 96.1% and specificity of 86.9%, was found. For PR the cytohistologic correlation was 71.2%, with a sensitivity of 65.7% and specificity of 83.8%. CONCLUSION: ICC determination of hormone receptors in routinely fixed smears obtained by FNAC is a simple method that correlates adequately with the results of IHC determinations, especially for ER. PMID- 11917584 TI - Diagnosis of B-cell lymphoma. Utility of the polymerase chain reaction for detecting clonality from archival cytologic smears. AB - OBJECTIVE: To apply the polymerase chain reaction (PCR) to detect clonality for potentially helping to establish a definitive diagnosis of lymphoma in cytologic material. STUDY DESIGN: In this retrospective study, Papanicolaou-stained cytologic smears and formalin-fixed, paraffin-embedded tissues from 17 cases of B cell lymphoma were examined to investigate their clonality by a PCR technique using three different approaches (FR3, FR3A and FR2) for amplification of immunoglobulin heavy chain genes. Cytologic smears from 10 cases of nonneoplastic lymphoid tissues and T-cell lymphomas served as negative controls. RESULTS: Monoclonality was detected in 9 of 17 cases (53%) of B-cell lymphoma in cytologic smears as compared with 8 of 16 cases (50%) in tissue sections. Semi-nested PCRs (FR3A/FR2) were superior to the single PCR (FR3) in the detection rate (41% vs. 18%). Five of seven cases (71%) of marginal zone B-cell lymphomas showed monoclonality, whereas only 4 of 10 cases (40%) of diffuse large B-cell lymphomas did so. Monoclonality was demonstrated in none of the negative controls. CONCLUSIONS: Clonality detection in B-cell lymphomas by PCR using cytologic smears is specific and equal in sensitivity to that using formalin-fixed, paraffin-embedded tissues. The detection rate is especially excellent in marginal zone B-cell lymphoma, in which the cytologic diagnosis is particularly challenging. Combined seminested PCRs for FR3A and FR2 are advocated for a reliable assessment of clonality. PMID- 11917583 TI - Cytologic evaluation of low grade transitional cell carcinoma and instrument artifact in bladder washings. AB - OBJECTIVE: To identify key cytologic features for the separation of low grade transitional cell carcinomas (TCCs) from nonneoplastic lesions in bladder washings. STUDY DESIGN: The cytomorphologic features of 95 bladder washing specimens showing papillary fragments, which included 50 low grade TCCs and 45 nonneoplastic lesions, were reviewed retrospectively. RESULTS: Bladder washings from low grade TCCs showed papillary and irregular groups of cells with ragged borders, cytoplasmic homogeneity and subtle nuclear changes, such as increased nuclear/cytoplasmic ratio and irregular nuclear border. Bladder washings after instrumentation from nonneoplastic lesions of the bladder showed cellular specimens with cohesive, ball-shaped and papillary clusters with smooth borders lined with a denser-staining cytoplasmic collar. Reactive urothelial cells often displayed loose aggregates with irregular borders but no cytoplasmic collar. CONCLUSION: In bladder washing cytology, nuclear changes and cytoplasmic homogeneity play a major role in the diagnosis of carcinoma. PMID- 11917585 TI - Kimura's disease. Diagnosis by aspiration cytology. AB - OBJECTIVE: To characterize the cytologic features of Kimura's disease and distinguish the differentiating features from those of other conditions with similar cytologic findings. STUDY DESIGN: In eight cases of Kimura's disease the cytologic features were correlated with the histologic findings. RESULTS: The cytologic features included a polymorphous lymphoid population with an admixture of significant numbers of eosinophils, fragments of collagenous tissue, endothelial cells and occasional polykaryocytes. CONCLUSION: Various conditions, both benign and malignant, may mimic Kimura's disease clinically and on smears. These have to be ruled out before making a diagnosis of Kimura's disease. The cytologic features of Kimura's disease have to be interpreted in the appropriate clinical setting in order to make a correct preoperative diagnosis. PMID- 11917587 TI - Cytologic findings of atypical adenosis of the breast. A case report. AB - BACKGROUND: Atypical apocrine adenosis, a well-described histopathologic entity, can sometimes be misdiagnosed as carcinoma. Apocrine cells can also appear atypical in cytopathology and be mistaken for carcinoma. Occasional case reports describe false positive cases due to the presence of apocrine cells in a few cases of radial scars and atypical apocrine metaplasia and in a degenerated cyst. CASE: A 37-year-old female underwent ultrasound-guided fine needle aspiration of an ill-defined breast nodule. The aspirate showed clusters and single cells containing abundant granular to focally vacuolated cytoplasm; enlarged, pleomorphic nuclei with irregular nuclear membranes; granular chromatin; and prominent nucleoli. These cells were distinct from and larger than the surrounding ductal and myoepithelial cells. Excision showed a nodular area of atypical apocrine adenosis adjacent to previous biopsy changes, correlating with the cytologic findings. CONCLUSION: Atypical apocrine adenosis can mimic carcinoma in histopathology and cytopathology. One should be cautious when reviewing apocrine cells in cytology, given their atypical features, especially their single, dispersed nature. However, the presence of accompanying benign cellular elements supports a benign diagnosis. Surgical biopsy should be recommended based on the cytologic findings. PMID- 11917586 TI - Usefulness of light microscopy in lymph node fine needle aspiration biopsy. AB - OBJECTIVE: To evaluate light microscopic examination of lymph node fine needle aspiration biopsy (FNAB) in order to determine the indications for ancillary procedures and biopsy. STUDY DESIGN: Reports and smears from 693 consecutive lymph node FNABs were reviewed. Fifty-five cases were excluded because of inadequacy of the material, and another 26 were excluded because follow-up information was not available. RESULTS: Cytologically, 220 cases were diagnosed as positive for malignancy and 392 as negative. Global sensitivity was 94.1% and specificity 96.9%. Sensitivity was higher for nonlymphoid neoplasms (98.2%) than for lymphoproliferative disorders (82.8%). CONCLUSION: Lymph node FNAB is a cost effective procedure, and with adequate cytologic examination and follow-up, a large number of biopsies and time-consuming ancillary techniques can be avoided. PMID- 11917588 TI - Diagnosis of a vulvar granular cell tumor by fine needle aspiration biopsy. A case report. AB - BACKGROUND: Granular cell tumor (GCT) (granular cell "myoblastoma") is an uncommon neoplasm that may mimic carcinoma both clinically and morphologically. Fine needle aspiration diagnosis of vulvar GNT has been described on only one prior occasion. CASE: A 74-year-old, black female presented with a mass in the left labia. Fine needle aspiration biopsy revealed rare intact cells; abundant, granular, cytoplasmic fragments; and bland, ovoid, stripped nuclei. The intact cells were arranged in loose aggregates. Each sampling was exquisitely painful to the patient despite the use of local anesthesia. CONCLUSION: Cytologists should be aware of the distinctive clinical and morphologic appearance of GCT. The cytologic findings of vulvar, GCT are identical to those described at other body sites. Definitive diagnosis before extirpation permits definitive therapy. PMID- 11917589 TI - Unusual cases of metastases to the breast. A report of 17 cases diagnosed by fine needle aspiration. AB - BACKGROUND: Although nonmammary tumors metastatic to the breast are relatively uncommon, a correct diagnosis is essential to appropriate management. Radiologically these lesions are single, round, discrete lesions without the spiculations of primary malignancies. Fine needle aspiration biopsy (FNAB) may provide a definitive diagnosis, thereby sparing patients unnecessary surgery. CASES: Seventeen cases of nonmammary malignancies diagnosed by fine needle aspiration of the breast were identified in the cytopathology files at three different institutions from 1989 to 1999. Three of the cases are of particular interest, including a mucoepidermoid carcinoma of salivary gland origin and a small cell undifferentiated carcinoma of rectal origin, neither of which has been reported in the literature previously. The third case was a male with a breast mass that was originally thought to be primary based on clinical, cytologic and immunocytochemical features but subsequently was determined to be a metastasis from the lung. CONCLUSION: Virtually any malignancy may metastasize to the breast. FNAB is the best approach to the diagnosis of tumors that either clinically or radiographically are not typical of primary breast tumors. Extramammary neoplasms metastatic to the breast may be definitively diagnosed by FNAB, resulting in the most appropriate as well as cost-effective patient management. PMID- 11917590 TI - Cytodiagnosis of erythema nodosum leprosum. A case report. AB - BACKGROUND: The application of cytology in leprosy has been restricted to the evaluation of morphologic and bacterial indices by slit skin smears to facilitate diagnosis of cases according to the Ridley-Jopling scale. Isolated reports have now documented the use of fine needle aspiration cytology (FNAC) in the diagnosis of leprosy. CASE: A 45-year-old male presented with the abrupt onset of multiple nodular eruptions all over the body. The clinical diagnosis was Sweet's syndrome. FNAC showed numerous neutrophils in a background of foamy macrophages. Special stains revealed the presence of a large number of fragmented acid-fast bacilli in the smears. A diagnosis of erythema nodosum leprosum (ENL) was made on FNAC. CONCLUSION: The presence of neutrophils in a characteristic milieu of foamy macrophages is seen in lesions of ENL. Such a picture should prompt the cytologist to use a modified Ziehl-Neelsen stain to demonstrate acid-fast bacilli, as ENL can present as an acute episode in patients without a previous diagnosis of leprosy. PMID- 11917591 TI - Fine needle aspiration biopsy of a posttransplant lymphoproliferative disorder with pronounced plasmacytic differentiation presenting in the face. A case report. AB - BACKGROUND: Posttransplant lymphoproliferative disorders (PTLDs) occur in fewer than 2% of transplant patients. However, as a group, 54% of PTLD patients die of these diseases. Presentation as only skin/superficial soft tissue nodules is rare, with this the second such reported case, and this is the only fine needle aspiration biopsy (FNAB) of such a case as well as the only FNAB of a plasmacytoid monomorphous/monoclonal PTLD. CASE: A 48-year-old, white male, seven years status post kidney transplantation, presented with a 2.5-cm mass in the skin/soft tissue anterior to the right maxillary sinus. FNAB showed a moderately cellular smear composed of discohesive cells, many with the morphology of plasma cells and some with the morphology of large lymphocytes. Flow cytometry showed these cells to be a monoclonal B-cell population, and a diagnosis of monomorphous/monoclonal PTLD was made. The diagnosis was subsequently confirmed by histology. The patient ultimately died. CONCLUSION: The clinical course of the present patient was grave as compared with the course of the other reported patient. PMID- 11917592 TI - Fine needle aspiration diagnosis of lipoblastoma of the parotid region. A case report. AB - BACKGROUND: Lipoblastomas are rare tumors of embryonal fat that occur in infants and children. They are usually located in the extremities and trunk. Two cases in the parotid region have been described. A diagnosis on fine needle aspiration (FNA) specimens has been reported in six cases. CASE: Lipoblastoma of the parotid region occurred in a 6-year-old boy and was diagnosed by FNA. Cytology showed rare lipoblasts and hibernomalike cells in a myxoid background with spindle and stellate mesenchymal cells, mature adipose cells and plexiform capillaries. A 7.0 cm, well-circumscribed mass with lobulated adipose tissue and delicate fibrous bands was resected. Microscopically, it showed a lobulated myxoid stroma, many capillaries, mesenchymal cells, lipoblasts and mature adipose cells. CONCLUSION: Lipoblastoma has to be differentiated from myxoid and lipomatous soft tissue tumors, especially from myxoid liposarcoma, a malignancy that classically affects older individuals and shows pleomorphism, atypical lipoblasts and chromosome-12 translocation. A lipoblastoma diagnosis must be established only after careful consideration of all available clinical, radiologic, cytogenetic and morphologic data. PMID- 11917593 TI - Anaplastic large cell lymphoma of the mediastinum diagnosed by transbronchial scratch cytology. A case report. AB - BACKGROUND: Anaplastic large cell lymphoma (ALCL) is a subtype of non-Hodgkin's lymphoma characterized by CD30 antigen-positive, large neoplastic cells. We describe a case of ALCL suggested by cytologic examination of the tumor cells obtained from bronchial scratch preparations. CASE: A 26-year-old woman had had a dry cough since November 1996. Chest radiography in May 1997 revealed an abnormal shadow in the mediastinum extending to the pulmonary hilar region. The patient was hospitalized in June 1997. Computed tomography revealed a neoplastic lesion in the anterior mediastinum invading the right lung. Transbronchial scratch cytology revealed large, atypical lymphoid cells expressing CD30 and CD3 on immunocytochemical examination. A transcutaneous mediastinal biopsy was performed and a diagnosis of ALCL made. CONCLUSION: Differentiation from Hodgkin's disease was the most difficult point in this case. Detailed cytologic observation and CD3 positive immunocytology led to the correct diagnosis. The cell transfer technique of Sherman et al was very useful for immunocytologic staining. Thus, transbronchial scratch cytology was an especially valuable and effective procedure in this case. PMID- 11917594 TI - Plasmacytoid urothelial carcinoma of the bladder. A case report and the first description of urinary cytology. AB - BACKGROUND: Plasmacytoid carcinomas are rare variants, and there are only a few reported examples of plasmacytoid carcinoma of the urinary bladder in the English language literature. We now report another such case and the first in which there was examination of urinary cytology. CASE: A 79-year-old, Caucasian woman who presented with gross hematuria following a revascularization procedure on the right arm was found to have an extensive, diffuse carcinoma of the bladder. On biopsy, there were single, round and polygonal tumor cells with a striking plasmacytoid appearance infiltrating diffusely throughout the edematous lamina propria. Immunocytochemical stains confirmed an epithelial classification, and carcinoma in situ was demonstrated in the contiguous urothelium. Voided urine cytology before and after cystoscopy and biopsy demonstrated large, dyshesive tumor cells with plasmacytoid features. CONCLUSION: A case of plasmacytoid variant of urothelial carcinoma of the bladder is reported, with the first description of its urinary cytology. PMID- 11917595 TI - Extraskeletal myxoid chondrosarcoma of the chest wall masquerading as a breast tumor. A case report. AB - BACKGROUND: Extraskeletal myxoid chondrosarcoma is a rare tumor that usually occurs in the soft tissues of extremities. Cytologic features of chondrosarcoma arising from a rib and presenting as a breast mass were reported by Molyneux et al in 1995. However, to the best of our knowledge, the cytology of extraskeletal myxoid chondrosarcoma of the chest wall presenting as a breast mass has not been documented before. CASE: A 30-year-old female presented with left-sided chest pain and a hard lump in the breast of two months' duration. Fine needle aspiration cytology was done, and a possible diagnosis of mucinous carcinoma or mixed tumor of the breast was suggested, with advice to prepare a frozen section before undertaking a radical procedure. On imprint cytology and frozen section a diagnosis of extraskeletal myxoid chondrosarcoma was made and confirmed by histopathology and immunohistochemistry. CONCLUSION: It is rare for extraskeletal myxoid chondrosarcoma to occur in the chest wall. This problem is compounded by the fact that cells of extraskeletal myxoid chondrosarcoma which resemble epithelial cells, can very closely mimic some malignant breast tumors. PMID- 11917597 TI - Squash cytology of intradural myxopapillary ependymoma. PMID- 11917596 TI - Metastatic papillary thyroid carcinoma in a child treated for congenital heart disease: diagnosis by fine needle aspiration cytology. PMID- 11917598 TI - Aspiration cytology of malignant hemangiopericytoma metastatic to the lung in a patient with previous primary malignant tumors: differential diagnosis. PMID- 11917599 TI - Preoperative cytologic diagnosis of clear cell follicular neoplasm of the thyroid. PMID- 11917600 TI - Rare cytologic findings in medullary thyroid carcinoma. PMID- 11917601 TI - Could Warthin's tumor become metaplastic after fine needle aspiration? PMID- 11917602 TI - Non-small cell carcinoma of the lung metastatic to the lower female genital tract and mimicking glassy cell carcinoma. PMID- 11917603 TI - Immunoreactivity of p53, PSA and bcl-2 in amniotic fluid smears from the second trimester of pregnancy. PMID- 11917604 TI - Warthin's tumor: metaplastic changes after fine needle aspiration biopsy. PMID- 11917605 TI - Growing leaders. Preparing the workforce for the future. PMID- 11917606 TI - Resistance to change. Causes and solutions. PMID- 11917607 TI - Merging trusts. PMID- 11917608 TI - Error making and nursing. PMID- 11917609 TI - Rehabilitation. Part 3: Setting up a service. PMID- 11917610 TI - If all these nurses have been recruited why does it feel as if things are little better in their clinical environments? PMID- 11917611 TI - Assessing staff performance in a rapidly changing service. PMID- 11917612 TI - Minority report. PMID- 11917613 TI - Better care for patients. PMID- 11917614 TI - Interprofessional education. PMID- 11917615 TI - Providing out of hours urgent care. PMID- 11917616 TI - Older patients in need of social worker care in A&E. PMID- 11917617 TI - Ambulance responses at a disaster site. PMID- 11917618 TI - Continuing Professional Development. Anaphylactic shock. AB - The causes of anaphylactic shock and presenting features are not clearly understood. In addition, the treatment of people suffering from anaphylactic or anaphylactoid reactions has not always been effectively provided. This article aims to rectify some of the current areas of concern, especially highlighting the recent work carried out by the Resuscitation Council (UK) on this subject (Resuscitation Council UK 1999/2001). After reading this article you should be able to: Explain the circulation of the blood and changes that occur during an anaphylactic reaction. Summarise the antigen/antibody mechanism especially in relation to allergic reactions. Describe the difference between anaphylactic and anaphylactoid reactions. Recognise the presenting signs and symptoms of anaphylaxis and shock. Describe the assessment and appropriate intervention for a patient suffering from an anaphylactic reaction. Explain what prevention/support services are available to suffers. PMID- 11917619 TI - Better child facilities but specialty ENs still rare. PMID- 11917620 TI - A question of supply and demand. PMID- 11917621 TI - Elaine's story. PMID- 11917622 TI - Localized scleroderma or morphea? AB - Morphea is a frequently mild, benign, and self-limiting skin disease with a less than 1% reported chance of progressing to systemic scleroderma. Morphea is a sufficient and less terrifying name for these disorders than localized scleroderma. PMID- 11917623 TI - Sports dermatology. AB - Sports-related dermatoses are among the most common ailments affecting athletes, from the neophyte to the professional. The general types of skin conditions related to athletic activity include infections, aberrant growths, inflammatory reactions, traumatic conditions, and environment-related conditions. Knowledge of the skin disorders common to athletes and familiarity with the specific activities of each sport which place these athletes at greater risk are critical to making accurate clinical diagnoses, and educating athletes regarding treatment interventions and prevention. PMID- 11917624 TI - What's your assessment? Basal cell carcinoma. AB - The "What's Your Assessment?" series includes a short case presentation and differential diagnosis. It is followed by a discussion of the disease or condition and the rationale used in each step of the assessment. PMID- 11917625 TI - Herpes simplex. AB - The "Clinical Snapshot" series provides a concise examination of a clinical presentation including history, treatment, patient education, and nursing measures. Using the format here, you are invited to submit your "Clinical Snapshot" to Dermatology Nursing. PMID- 11917626 TI - Wound assessment and evaluation. AB - The "Wound Assessment and Evaluation" series includes a short case presentation and differential diagnosis. It is followed by a discussion of the disease or condition and the rationale used in each step of the assessment. PMID- 11917627 TI - Tattoos, piercings, and skin adornments. AB - Tattoos, body piercings, and other skin adornments have a long and fascinating history, and continue to be prominent today. It is important for dermatology nurses to understand and to educate their patients regarding the dermatologic implications of tattooing, body piecing, and scarification. PMID- 11917628 TI - No known link between sunscreen eye exposure and blindness. PMID- 11917629 TI - New registry will benefit patients with antiphospholipid syndrome. PMID- 11917630 TI - Breakthrough treatment for improving color of scars and uneven skin pigment. PMID- 11917631 TI - Laser therapy and microdermabrasion treat acne scars. PMID- 11917632 TI - [Huriez' law and medical imaging]. PMID- 11917633 TI - [Studies of cost-effectiveness and cost-usefulness in radiology]. AB - In the current context of significant increase of health care costs over the last decades, and in a system of global budget for health care, the concept of cost effectiveness is one of the leading elements in the political decision making process for a given strategy. Therefore, it is important for the physician to be able to understand and critically interpret cost-effectiveness and cost-utility analyses. This article tries to illustrate comprehensively some of their key concepts. The perspective and the time horizon of the study should be clearly specified. The cost-effectiveness ratio is a synthetic summary based on a micro costing approach in order to determine the true costs (numerator), and on an effectiveness (utility) assessment which should take into account the preferences of the community (denominator) in order to allow comparisons between interventions of different natures. Advances in the development of decision analysis softwares and in the standardization of the methodology of these studies have yielded considerable improvement in the reliability of their results. Several persisting methodological problems are the scope of current research, such as the discounting rate and the calculation of the minimal sample size required to reach a statistically significant threshold. PMID- 11917634 TI - [Decision analysis in radiology]. AB - Decision analysis is a technique which allows to clarify in an explicit, probabilistic and quantitative way the possible answers to a problem and to help the decision making process. The creation of algorithms, graphically displayed as decision trees in most cases, requires the introduction of quantitative information of two types: probabilities of the events that result from answering to the initial question, and utilities of the possible outcomes of these events. The choice of the optimal solution is based on the calculation of combinations of these data. Although the construction of models can be complex and time consuming, their practical use has been simplified by the ease and user friendliness of available softwares. To date, the applications of decision analysis in medicine have focused on cost-effectiveness studies and on the simulation of randomized trials. At the individual level, clinical applications of decision analysis could provide in a near future an interesting tool for the clinician involved in patient management. Even when doubts remain regarding the reliability of the data entered into the model, further sensitivity analysis provide a convenient way to test the validity of the drawn conclusions. This article hopefully offers a simplified and practical approach to basic decision analysis. PMID- 11917635 TI - [Imaging of the anterior abdominal wall: ultrasonography and CT features]. AB - US and CT have both become major tools in imaging of the anterior abdominal wall. The goal of this pictorial review is to illustrate the respective roles of US and CT in the evaluation of normal and abnormal anterior abdominal wall. PMID- 11917636 TI - [Imaging of the hip in healthy children in MRI: evaluation of the acetabular coverage]. AB - PURPOSE: To define with MR imaging a reference value for acetabular coverage on coronal and sagittal images for a population of healthy children. MATERIALS AND METHODS: 36 children (72 hips) where prospectively studied over 1 year. T1 weighted spin-echo images in both coronal and sagittal planes were obtained. Slice thickness was 4 mm. Measurement of acetabular coverage (AHI: Acetabular Head Index) was made in both planes with two different landmarks: bone and cartilage. Study of the evolution with age was made. RESULTS: The acetabular coverage is symmetrical and decreases with age. Its measurement is reproducible. CONCLUSION: This study shows that measurement of acetabular coverage (AHI) is easily obtained at MR imaging and is reproducible. We suggest 83 (mean 1SD) as the inferior limit for AHI based on cartilage measurements. We introduce sagittal values, never published. We believe that MR, by its ability to demonstrate the cartilaginous surfaces, will play a major role in the true understanding of the anatomical relationship between the femoral head and the acetabulum. PMID- 11917637 TI - [Publication rate of original papers orally presented at the Journees Francaises de Radiologie 1996]. AB - OBJECTIVE: The purpose of the present study was to determine the publication rate in Medline-indexed journals of papers originally presented at the Journees Francaises de Radiologie (JFR96). MATERIALS AND METHODS: Proceedings from the JFR 96 were reviewed by two observers in conference. A Medline search encompassing 1997-1999 was performed for the lead author of all abstracts of original papers. Publication year and the journal of publication were recorded. RESULTS: 456 oral presentations were analysed, 39 papers were subsequently published corresponding to a publication rate of 8.5%. 10 papers were published in Radiology, 4 in European Radiology and 4 in the Journal de Radiologie, the other 21 in other journals. Publication rate was significantly higher for two sessions: Research session (publication rate was 22%) and Bone and Joint session (publication rate was 16%). CONCLUSION: Less than 10% of abstracts presented at JFR 96 were subsequently published in Medline-indexed journals. This rate is lower than the publication rate of papers presented at other medical meetings. PMID- 11917638 TI - [Leiomyosarcoma and stromal tumor of the pancreas]. AB - A tumor of the head of pancreas was an incidental finding on US of two patients aged 52 and 61 years presenting with abdominal pain. The tumor was studied by computed tomography, MRI and endoscopic ultrasonography. Surgical biopsy of a liver lesion was performed in one case and partial duodenopancreatectomy was performed in the other case. Histological evaluation, including immunohistochemistry, showed leiomyosarcoma and stromal tumor respectively. Whatever the site of origin, most mesenchymal pancreatic tumors are hypervascular, heterogeneous and have a necrotic center. PMID- 11917639 TI - [Bilateral and spontaneous hemorrhage of the renal sinus. Report of a case]. AB - Renal sinus hematoma is a rare complication of anticoagulant therapy. Prognosis is usually excellent with spontaneous regression. The authors report a case of spontaneous bilateral renal sinus hematoma in a 59-year-old woman, treated by anticoagulants for a cardiac disease. Diagnosis and follow-up are made using computed tomography. PMID- 11917640 TI - [Plasmacytoma and AIDS: unusual duodenal localization]. AB - We report an uncommon presentation of plasmocytoma in an AIDS patient. AIDS is associated with an increased risk of neoplasms. The incidence of Plasma Cell tumors in HIV-positive patients is greater than in non-infected patients. Multiple factors contribute to B cell neoplasms development. PMID- 11917642 TI - [Revision of the law of December 20, 198: consensus platform of propositions from professional societies, agencies, and patient associations]. PMID- 11917641 TI - [Spontaneous resolution of asymptomatic false aneurysm of the pulmonary artery induced by Swan-Ganz catheter]. AB - A pulmonary artery pseudoaneurysm developed after placement of a Swan-Ganz catheter for liver surgery is described. This observation is unusual for two reasons. Pulmonary artery pseudoaneurysm was discovered on a simple post operative chest radiograph. Spontaneous thrombosis and radiographic regression were observed. PMID- 11917643 TI - [Quid? Hepatic hydatid cyst ruptured into the thorax]. PMID- 11917644 TI - [Quid? Bone hydatidosis]. PMID- 11917645 TI - [Imaging in gynecology]. PMID- 11917646 TI - [Imaging of the female pelvis in adolescence]. AB - The aim of this chapter is to familiarize the radiologists with the evolutive morphological features of the female genital apparatus, as it can be seen during the puberty, and to view the pathologies panel which may occur in ths period of life. This chapter is going to illustrate the morphological sonographic modifications from the childhood to the adolescence, emphasing the signs of pubertal maturation (uterus and ovaries shape, size and vascularization). Through clinical signs (delayed sexual maturation, primary or secondary amenorrhea, menstrual dysfunction, acute, cyclic, chronic pelvic pain and pelvic mass) the different pathologies are viewed, illustrating the important role of ultrasonography, but not an exclusive role. PMID- 11917647 TI - [Clinical case: questions--answers. Imaging of the female pelvis in adolescence]. PMID- 11917648 TI - [Imaging of gynecologic malformations]. AB - Female genital tract anomalies are common (1 to 2% of the female population), and may lead to multiple clinical manifestations: amenorrhea, infertility, spontaneous repeated miscarriage, pelvic pain, endometriosis. They are caused by intra-uterine insults between weeks 6 and 18 of gestation. They are classified according to their embryologic origin. Imaging relies essentially on ultrasound and MRI, and indications for hysterosalpingography are less common. Imaging must classify the malformation and detect complications in order to assess the fertility prognosis and treat complications. PMID- 11917649 TI - [Clinical case: questions--answers. Imaging of gynecologic malformations]. PMID- 11917650 TI - [Imaging of the endometrium]. AB - During genital activity, physiological and pathological modifications can be observed; Pre- and postmenopausal menometror-rhagia are the principle clinical signs of various endometrial pathologies: benign (polyp, atrophy or endometrial hypertrophy), malignant (cervical or endometrial carcinoma) or neighboring pathologies (myometrium or ovary). The value and methods of various imaging techniques (B-mode and Doppler abdominal and endovaginal ultrasonography, hysterosonography, computed tomography, MR imaging and hysteroscopy) are described together with symptomatological features permitting identification of the endometrial pathology. PMID- 11917651 TI - [Clinical case: questions--answers. Imaging of the endometrium]. PMID- 11917652 TI - [Myometrium diseases]. AB - Leiomyoma and adenomyosis are the most frequent myometrial disorders, followed by cystic and vascular disorders. After an update on the anatomy, physiology and histology of the myometrium, the authors will describe the normal sonographic and MRI aspects of the uterus. The pathology of leiomyoma will be recalled, and the various forms of myoma encountered during Doppler US, hysterography, hysterosonography, MRI and computed tomography will be described. The therapeutic use of pelvic arteriography with arterial embolization will be discussed. Adenomyosis is the second most frequent essential myometrial disorder, and is often associated with leiomyoma. After giving a histopathologic definition, the authors will examine in some detail the diagnostic value and limitations of transabdominal, pelvic and endovaginal sonography and MRI. Intramyometrial cystic disorders (cystic adenomyosis, myoma with cystic degeneration and vestigial cysts) and vascular disorders (intramyometrial and parametrial vascular malformations) are rare, but a sound knowledge is required to optimize their management, which is based on surgery and interventional vascular techniques. PMID- 11917653 TI - [Clinical case: questions--answers. Myometrium diseases]. PMID- 11917654 TI - [Imaging of the cervix and the vagina]. AB - US and MRI currently are the best imaging modalities to evaluate pathology of the uterine cervix and vagina. Carcinoma of the cervix is the most frequent indication for imaging. MRI allows preoperative staging of cervical carcinoma based on FIGO classification, and post treatment follow-up. Other uterine cervix diseases are less frequently imaged and include a wide range of entities that most frequently cause increased T2W signal at MR imaging. Pathology of the pelvic floor, vagina, vulva, and perineum also includes a wide range of entities that have seldom been described in the imaging literature. PMID- 11917655 TI - [Clinical case: questions--answers. Imaging of the cervix and the vagina]. PMID- 11917656 TI - [Tubal diseases: from diagnosis to intervention]. AB - Ninety years after it was first performed, hysterosalpingography remains one of the most important tests in helping an infertile couple decide between numerous treatment options. The application of angiographic techniques to hysterosalpingography has revolutionized diagnosis and treatment of proximal tubal obstruction. Results from worldwide centers have shown that nonsurgical catheter recanalization of proximally obstructed tubes can be accomplished in up to 90% of patients using standard techniques. Pregnancy incidence in a population who was recommended for tubal microsurgery or in vitro fertilization, but who underwent catheter tubal recanalization instead without any other therapy, was 58% at one year and all pregnancies were intrauterine. Fluoroscopic fallopian tube catheterization should be the first treatment for proximal tubal obstruction, with the more expensive and invasive procedures reserved for the small number of women who fail the radiologic procedure. PMID- 11917657 TI - [Clinical case: question--answers. Tubal disease: from diagnosis to intervention]. PMID- 11917658 TI - [Pelvic endometriosis]. AB - Endometriosis is a common disease that affects about 20% of women of childbearing age. At transabdominal and endovaginal ultrasound, in 90% of cases, ovarian endometriosis appears as a homogeneous lesion with low level echoes most often associated with clots. However this modality is insufficient to detect associated lesions. MRI is as accurate as ultrasound to detect ovarian endometriosis which most often appears with a signal higher than that of adipose tissue on T1W images and a bright signal area on fat suppressed images. The main advantage of this examination is not only to detect small ovarian implants but to also detect associated subperitoneal implants. These lesions involving the bladder as well as posterior structures such as the uterosacral ligaments, rectovaginal septum, and rectum appear as fibromuscular masses containing hyperintense signal areas on T1W images, or as purely fibrotic lesions that can be quite difficult to diagnose. Laparoscopy remains the procedure of choice to detect peritoneal implants. PMID- 11917659 TI - [Aspirin-ACE inhibitors interaction: myth or reality?]. PMID- 11917660 TI - [The crisis of contemporary medicine or Hippocrates' "second death"]. PMID- 11917661 TI - [Isolated iliac deep venous thrombosis. Study of 48 cases seen in 7 years among 18,297 echo-Doppler evaluations of the lower limbs]. AB - Isolated iliac venous thrombosis (IIVT) is uncommon. Duplex ultrasonography of the iliac vessels is not recommended and not generally performed. OBJECTIVE: The purpose of this study was to determine the frequency of IIVT in a hospital recruitment population and to identify characteristic features of onset which might be associated with this localization in order to better target explorations. MATERIAL AND METHODS: The study included 18,297 patients referred for Duplex-ultrasonographic exploration of possible deep vein thrombosis of the lower limbs between January 1st 1994 and December 31st 2000. Selection of isolated iliac thrombosis, defined as the absence of retrograde extension to the common femoral vein, was made from the digitalized data recorded daily. The following factors were tested: sex, age, absence of clinical signs in the lower limb, presence of pulmonary signs. The raw odds ratios were calculated followed by construction of a multivariate logistic regression model. The circumstances of onset were retrieved from the patient's medical files. RESULTS: Isolated iliac venous thrombosis was discovered in 48 patients, i.e. 0.26% (95% CI 0.19%-0.35%) in the recruitment population and 0.82% (95% CI 0.61%-1.09%) among the 5827 patients with thrombosis. The common iliac was involved predominantly (35 out of 48). The left side predominated in women compared with men (24/36 versus 4/12) (p = 0.04). Specifically female circumstances (oral contraceptives, peri-obstetrical period) always led to a left localization. For the other identified circumstances (cancer, inflammatory bowel disease, orthopedic surgery, pelvic trauma), there was no predominant side. Variables explaining the multivariate model were sex, age (less than or more than 35 years), suspected pulmonary embolism, and age interaction with suspected pulmonary embolism. For women, the risk of IIVT was twice as high as for men (OR = 1.97, 95% CI 1.02-3.81). Young age was also a risk factor for IIVT and increased with suspected pulmonary embolism. The odds ratio for subjects under 35 varied from 4.20 (95% CI 1.79-9.84) without suspected pulmonary embolism to 35.01 (95% CI 14.78-82.89) with suspected pulmonary embolism. CONCLUSION: The incidence of isolated iliac venous thrombosis is very low (0.26) in this hospital recruitment population but reached 9.40% in young women under 35 with suspected pulmonary embolism. These age and sex characteristics are also the principal circumstances for the development of these thrombotic events (pregnancy, post partum period, oral contraception). Under these circumstances, it is necessary to carefully explore the iliac vessels with duplex ultrasonography. PMID- 11917662 TI - [Endoluminal treatment of peripheral aneurysm with covered endoprosthesis]. AB - PURPOSE: To evaluate the feasibility and middle term patency results, for endovascular treatment of peripheral aneurysms, using covered stents. MATERIALS AND METHODS: Between December 1993 and may 2000 25 peripheral aneurysms on 24 patients, mean age 67.8 years (42-81), were treated with covered stents (Cragg EndoPro system or passenger, hemobahn, Word Medical). Aneurysm locations were: 17 iliac, 4 popliteal, 1 femoral, 1 subclavian, 1 carotid. The follow-up range from 1 to 75 months (mean 30.1 months). RESULTS: Successful aneurysm exclusion was achieved in all patients, while maintaining good lower extremity perfusion. All but one stent remained patent during the early follow-up period. At one year 94.4% stents were patent, at two years 90.2. CONCLUSION: Treatment of peripheral aneurysms with covered stents has a high rate of immediate procedural and clinical success. At middle term follow-up patency is encouraging except with popliteal procedure, however long term follow-up and larger series is warranted to assess the place of this procedure as an alternative to the surgery. PMID- 11917663 TI - [Revascularization of the lower limbs with extra-anatomic shunts. Report of 80 cases]. AB - The extra anatomical bypasses are those which do not follow the usual arterial pathway. The aim of this study is to define the indication of extra anatomical bypasses and to compare them with other revascularisation procedures. In this paper we report a retrospective study carried between January 1988 and December 1999 involving 80 patients (78 men and 2 women) who underwent extra anatomical bypasses for revascularisation of their lower limbs (83 bypasses). The mean age of the patients was 62 years. The indication for revascularisation was chronic arteritis of the lower limbs in 72 patients (90%), infected aneurysms in 6 cases (7.5%) and acute aortic dissection in 2 cases (2.5%). We performed an axillo unifemoral bypass in 23 cases, an axillo-bifemoral in 17, a crossover ilio femoral or femoro-femoral bypass in 41 cases, and 2 transobturator ilio popliteal bypass right and left in 1 case. Hospital mortality rate was 10%. Hospital morbidity was about 65%. Limb salvage was 78%. Sixty patients were regularly followed during a mean follow-up of 31 months. Late mortality was 10%. In the end of our study global limb salvage rate was 74%. IN CONCLUSION: extra-anatomical revascularization allows acceptable limb salvage rate along with lower operative risk even in poor state patients. PMID- 11917664 TI - [Arterial aneurysm in Behcet's disease. Report of 5 cases]. AB - Behcet's disease is a systemic vasculitis generally involving the venous system. Arterial manifestations, usually aneurysm or more rarely occlusion, are less common. We analysed 5 cases of Behcet's disease with arterial aneurysm complications. There were 4 men and 1 woman, mean age 34.6 years. Mean delay to arterial complications was 7.8 years after the first sign of the disease. Four patients showed evidence aneurysms of the pulmonary arteries. One patient developed an aneurysm of the aortic and iliac artery. Phlebitis was associated with arterial involvement in 4 patients. Combined corticosteroid and cyclophosphamide therapy enabled regression of pulmonary aneurysms in 2 patients. One patient was operated, outcome was favorable. Histology showed fragmentation of the media associated with vasculitis. Two patients with pulmonary aneurysms died of massive hemoptysis. PMID- 11917665 TI - [Cutaneous angiosarcoma of the breast after treatment with conservative surgery and radiotherapy of breast carcinoma]. AB - Secondary cutaneous angiosarcoma of the breast is uncommon. This tumor develops after conservative treatment of breast cancer. We present a case that developed seven years after conservative treatment with surgery and radiotherapy for breast cancer. Several factors may be implicated in the pathogenesis of this tumor (carcinogenic effect of radiotherapy, HHV8 infection), but the essential factor appears to be chronic edema. Prognosis is poor but initial wide surgical resection can achieve prolonged remission. Early diagnosis and treatment are the main prognostic factors. PMID- 11917666 TI - [Clinical and plethysmographic changes after phlebologic thermal care in chronic venous insufficiency of the lower limbs]. PMID- 11917667 TI - [Changes in VCAM-1 cell adhesion molecule after phlebologic thermal care in chronic venous insufficiency of the lower limbs]. PMID- 11917668 TI - Review and analysis of evaluation measures used in nutrition education intervention research. AB - The purpose of this review is to provide a summary of the kinds of evaluation measures used in 265 nutrition education intervention studies conducted between 1980 and 1999 and an analysis of psychometric issues arising from such a review. The data are summarized in terms of tables for interventions with each of six key population groups: preschool children, school-aged children, adults, pregnant women and breast-feeding promotion, older adults, and inservice preparation of professionals and paraprofessionals. Measures evaluating knowledge and skills or behavioral capabilities were most widely used in studies with preschool, school aged, and inservice populations (50%-85%) and less widely used in studies with the other groups, particularly breast-feeding promotion (5%). Measures of potential psychosocial mediators or correlates of behavior such as outcome expectancies, self-efficacy, or behavioral intention were used in 90% of behaviorally focused studies with school-aged children and in about 20% of studies with adults. Dietary intake measures were used in almost all studies, primarily food recalls, records, and quantitative food frequency questionnaires. Short frequency instruments involving only foods targeted in the intervention such as fruits and vegetables are increasingly being used. Measures of specific observable behaviors are also increasingly being used. Physiologic parameters were used in about 33% of behaviorally focused interventions with school-aged children and adults, 20% with older adults, and 65% with pregnant women and/or their infants. Criterion validity of newly developed intake instruments and content validity of instruments measuring mediating variables were reported in the majority (range 50%-90%) of studies. Reliability and stability of measures of mediating variables were reported in 50% to 75% of studies, with reliability coefficients mostly about .6 to .7. Two major conclusions from this review are that evaluation measures should be appropriate to the purpose, duration, and power of the intervention and that measures should have adequate validity and reliability in relation to both the outcomes and characteristics of the target audience. Major implications are that considerable preliminary work needs to be done before any intervention study to develop and test evaluation instruments so that they are appropriate and have adequate psychometric properties, and cognitive testing of published instruments with each new target audience is essential. We will then be better able to make judgments about the effectiveness of nutrition education. PMID- 11917669 TI - Cost-benefit analysis indicates the positive economic benefits of the Expanded Food and Nutrition Education Program related to chronic disease prevention. AB - OBJECTIVE: To provide an estimated cost-benefit ratio for the Expanded Food and Nutrition Education Program (EFNEP), based on potential prevention of diet related chronic diseases and conditions. DESIGN: A retrospective cost-benefit study using demographic, food/nutrient intake, and food-related behavioral data previously collected on program participants by trained paraprofessionals, before and after an intervention. Actual costs of implementing EFNEP for 1 year (1996) were also used. SUBJECTS/SETTINGS: 3100 female and male adults who had participated in the Virginia EFNEP during 1996. INTERVENTION: Prior participation in 6 to 12 food/nutrition education lessons with subsequent graduation from EFNEP. MAIN OUTCOME MEASURES: Cost-benefit ratios for EFNEP, based on original assumptions and subsequent sensitivity analyses. STATISTICAL ANALYSIS: Program implementation costs were compiled and compared with monetized benefits of disease prevention to produce benefit-to-cost ratios. Excel and SPSS computer programs were used to compute cost-benefit ratios based on standard procedures used in the field of economics. RESULTS: The initial benefit-to-cost ratio was $10.64/$1.00, with subsequent sensitivity analyses producing ratios ranging from $2.66/1.00 to $17.04/1.00. IMPLICATIONS: The results of EFNEP intervention translate into a positive cost-benefit based on potential prevention of diet related chronic diseases and conditions. Thus, EFNEP is a good use of federal tax dollars. PMID- 11917670 TI - Focus groups on cardiovascular health in rural women: implications for practice. AB - OBJECTIVE: To determine rural women's perceptions about cardiovascular disease (CVD) prevention and behavior change for cardiovascular health. DESIGN: A trained moderator and nutritionist (observer) led the groups. The discussion guide was developed from the Health Belief Model and Social Cognitive Theory. The data were analyzed inductively looking at patterns and themes that emerged from the data and deductively looking at relationships from the theoretical models. SUBJECTS: Thirty-four white women participated in six groups: three younger (20-40 years; n = 18) and three older (40-55 years; n = 16) groups. Women with no more than 13 years of education were recruited by social service providers, Extension agents, and church groups. RESULTS: Rural women were unaware of their personal CVD risks. Common themes included the overriding influence of family preferences and cultural food patterns on women's food choices and the lack of support for adoption of a heart-healthy diet. Self-efficacy for dietary change spanned the continuum from no confidence (younger women) to complete empowerment (older women). CONCLUSIONS: Efforts need to be directed toward providing education and skill building for CVD prevention based on a better understanding of women's cultural beliefs and life situations. All women believed that dietary choices were important for cardiovascular health; however, they lacked the skills for food selection and preparation. Family preference and support are key to the adoption and maintenance of a heart-healthy eating plan. For interventions, women preferred active learning (hands-on experiences) coupled with group classes for learning and support. Rural women lacked these resources or access to a nutritionist. PMID- 11917671 TI - Children's perceived benefits and barriers in relation to eating breakfast in schools with or without Universal School Breakfast. AB - OBJECTIVE: The purpose of this study was to identify and compare perceived benefits and barriers related to breakfast consumption and concerns about weight among children in schools with or without a Universal School Breakfast Program (USBP). DESIGN: Teacher-administered survey at the end of a 3-year pilot program. SUBJECTS/SETTINGS: Fourth-, fifth-, and sixth-grade students in six USBP pilot schools (n = 827) and four control schools (n = 615). Control and pilot sites were matched by geographic location and socioeconomic status of students. Response rates were > 70%. VARIABLES MEASURED: Perceptions of benefits and barriers related to breakfast consumption and weight-related concerns. STATISTICAL ANALYSES PERFORMED: Chi-square tests were used to assess statistical differences in categorical responses to survey items. RESULTS: The majority of students perceived that eating breakfast provides benefits of increased energy and ability to pay attention in school. Commonly held perceptions of barriers to eating breakfast were lack of time and not being hungry in the morning. Compared with children in non-USBP schools, those in the USBP schools were less likely to wish they were thinner, to go on a diet, or skip breakfast because it might make them fat and more likely to believe that eating breakfast will give them energy and help them pay attention. IMPLICATIONS: Based on the results of this study, nutrition educators may find it helpful to develop educational materials and programs based on the reciprocal determinism construct of Social Learning Theory to promote breakfast consumption. The focus should be on practical strategies to address barriers and encourage behavioral changes for both children and their parents. PMID- 11917672 TI - Nutrition education materials from the National Institutes of Health: development, review, and availability. AB - Many of the institutes, centers, and offices (ICOs) within the National Institutes of Health (NIH) develop and disseminate nutrition education materials for the general public. These materials provide information about the relationship of diet to health and about associations between diet and specific diseases. The materials, which are drafted by the NIH or contract science writers, go through pretesting (for literacy level and appropriateness for target audiences) and ICO clearance (for scientific accuracy). To further ensure scientific and technical accuracy and consistency with the Dietary Guidelines for Americans, the materials then go through a two-tiered governmental review system. The first review is through the Nutrition Education Subcommittee (NES) of the NIH Nutrition Coordinating Committee. The second review, which is required for federal nutrition education materials, is conducted jointly by the Department of Health and Human Services (DHHS) Nutrition Policy Board Committee on Dietary Guidance and by the US Department of Agriculture (USDA) Dietary Guidance Working Group. The review process helps ensure consistency in nutrition messages within the NIH ICOs and among government agencies. The pretesting, ICO clearance, NES review, and joint DHHS/USDA review result in materials for nutrition educators that are high in quality, low in cost or free, easily accessible, appropriate for the intended target audience, and consistent with the Dietary Guidelines for Americans. PMID- 11917673 TI - Feed Your Mind: an interactive nutrition evaluation for teenagers. PMID- 11917674 TI - Team nutrition: a collaborative approach. PMID- 11917675 TI - [Radiologic diagnosis of lunate necrosis]. AB - In the literature little is to be found concerning diagnosis and differential diagnosis of Kienbock's disease. Because of technical development in radiological imaging, we now possess very detailed pictures. However, there are many radiological findings within the lunate bone, which can be misinterpreted as Kienbock's disease. Pathological findings of the lunate bone are demonstrated, which are not Kienbock's disease. These findings were compared to different stages of Kienbock's disease and identified. The early stages of Kienbock's disease show changes especially in the MRI, which are very similar to acute bone bruise, ulnar impaction syndrome, synovialitis or gout. Vessel tubes or an intraosseous ganglion can be mistaken for Kienbock's disease. In the late stages of Kienbock's disease, it can be difficult to differentiate this from pseudarthrosis of the lunate bone. The differentiation between Kienbock's disease and other pathological findings is at times very difficult--even with MRI. However, sometimes diagnosis can only be verified by MRI. We therefore suggest that Kienbock's disease must be correctly diagnosed with help of the MRI before commencing with any therapeutic steps. PMID- 11917676 TI - [Treatment of difficult scaphoid pseudarthroses by bone transplantation and additional implantation of the second dorsal metacarpal artery using the Fernandez and Eggli technique--report of experiences]. AB - One of the procedures for the treatment of scaphoid-nonunion with loss of blood supply to the proximal fragment involves the technique described by Fernandez and Eggli (1995). After resection of the necrotic bone and inserting a corticocancellous bone graft from the iliac crest, the second dorsal intermetacarpal artery is implanted into the proximal fragment of the scaphoid. In our hospital, six patients underwent this procedure from May 1998 to June 2000 because of necrosis of the proximal pole or an unsuccessful previous operation. Union was achieved in five patients. In one case, nonunion persisted and the cystic area increased. The described technique is suitable for improvement of bone healing of the scaphoid in complicated cases. The advantages of this technique lie in the predictable anatomy of the second intermetacarpal artery and the straightforward dissection. PMID- 11917677 TI - [Possible applications of pedicled vascularized bone transplants of the distal radius]. AB - Reverse-flow pedicle vascularized bone grafts (VBGs) from the distal radius may be used for the surgical treatment of carpal fracture nonunions and avascular necrosis. Such grafts remain viable with quantifiable blood flow as demonstrated in a recent canine experiment. In this paper, the vascular anatomy of the distal radius is demonstrated and the surgical technique of VBG harvest is described. Our indications and results for treatment of scaphoid nonunion and Kienbock's disease are discussed and compared with the current literature. Fifteen patients with scaphoid nonunion and nine patients with Kienbock's disease stage IIIa were treated by reverse-flow pedicled VBG. Range of motion and grip strength were measured postoperatively and compared to the contralateral hand in all patients. An outcome questionnaire was used to measure patient's satisfaction and ADL's in the patients with Kienbock's disease. The radiologic diagnostic was performed with conventional X-rays, trispiral tomograms and MRI. In the scaphoid nonunion group, all patients were male, with an average age of 27.6 years. Five patients had prior attempts of autogenous bone grafting which failed. Six patients had a radiographic suggestion of proximal fragment avascular necrosis. All scaphoids healed. Time to union was 11.1 weeks on average. Average follow-up was 36.2 months. Pain relief, range of motion and grip strength were very promising. The treatment of Kienbock's disease was also promising in nine patients. Mean patient age was 31 years, and follow-up averaged 32 months. Six patients had complete pain relief, and three had occasional pain. Grip strength reached 86% of the contralateral side (a 25% improvement from preoperative values). Postoperative MRI demonstrated progressive revascularization with time. The results of treating scaphoid nonunions are promising. The use of vascularized bone graft facilitates rapid, reliable union of established scaphoid nonunion, even with proximal location and/or avascular changes. Clinical results in Kienbock's disease have been gratifying as well, with MRI evidence of gradual revascularization on follow up studies. PMID- 11917678 TI - [Dorsal V-ligament reconstruction in scapholunate instability]. AB - In dynamic and static scapholunate instability after trauma, the repair of the scapholunate ligament is important to avoid scapholunate advanced collapse of the wrist. Direct suture even of fresh-torn ligaments can be technically demanding and occasionally unreliable, thus reconstruction may require additional tissue beside the ligament. Eighteen patients suffering from dynamic (n = 10) and static (n = 8) scapholunate instability were treated by a dorsal ligament reconstruction six months after trauma. A clinical wrist score according to Cooney showed an average of 86 points (maximum 100) within a follow-up of nineteen months after surgery. X-ray films documented no significant loss of scapholunate reduction. Using local tissue only, this method is always possible, very reliable and easy to perform. PMID- 11917679 TI - [Functional results of medio-carpal partial arthrodesis with excision of the scaphoid]. AB - Midcarpal arthrodesis with excision of the scaphoid for the treatment of painful carpal collapse has been performed in our hospital since 1993. A clinical study was carried out to evaluate the results and determine special factors, which might influence the results. 26 out of 29 patients operated until 1999 were reexamined after an average follow-up of 27 months. The results were evaluated according to the Mayo-modified wrist score. The DASH score and the pain disability index (PDI) were calculated postoperatively. Standard X-rays of the wrist were analyzed for alteration of the radio-lunate joint space, the position of the lunate with respect to the radius and the correction of the carpal height as calculated by the Youm index. Carpal collapse was due to scapholunate pathology (SLAC) in 12 cases, long-standing scaphoid nonunion (SNAC) in ten cases, perilunate dislocation, which was only simply reduced, in three cases, and calcium pyrophosphate deposition disease in two cases. The Mayo-modified wrist score improved significantly from an average of 46 points before to 76 points after surgery. The DASH score postoperatively was 22, the PDI 13. All patients reported improvement of their situation after the operation, eight were completely free of pain. The average range of motion from extension to flexion was 64 degrees, which was almost identical to the average preoperative value. The average grip strength before surgery was 24 kg, after surgery 34 kg. There was a correlation between the position of the lunate to the radius in the lateral X-ray and the range of extension. If the lunate was positioned correctly, wrist extension was significantly better. The radiolunate joint space was maintained during the period of observation with only two exceptions. Apparent subchondral sclerosis was seen in most of the cases. It was not possible to restore carpal height completely. Some results after perilunate luxations and one case of calcium pyrophosphate deposition disease were unsatisfactory. Midcarpal arthrodesis with scaphoid excision is a reliable method for treating radioscaphoid arthrosis caused by carpal collapse. Correction of the hyperextended position of the lunate is important to obtain optimum results. In our hospital, a wrist arthrodesis is only rarely performed nowadays, except in the rheumatoid patient. PMID- 11917680 TI - [Is chronic, untreated scapho-trapezoid arthrosis after resection arthroplasty of the carpometacarpal joint clinically relevant?]. AB - Basal joint arthrosis and scapho-trapezio-trapezoid arthrosis (triscaphe arthrosis) are common degenerative diseases with proven surgical treatment. Besides the extirpation of the trapezium during the resection-suspension arthroplasty, we have performed additional fusion of the scaphoid and the trapezoid (ST arthrodesis) in patients with heavy arthrotic changes in both locations. But the combination of these two surgical procedures results in a prolonged rehabilitation and may lead to more complications, e.g. nonunion. Thus the question arises, whether there is any clinical relevance to an untreated scapho-trapezoid arthrosis after resection-suspension arthroplasty of the carpometacarpal joint of the thumb. From 1992 to 1998, we performed resection suspension arthroplasty of the carpometacarpal joint of the thumb without additional intervention at the scapho-trapezoid joint in 229 patients. In reviewing the perioperative X-rays of these patients, we found 55 cases with untreated arthrosis of the scapho-trapezoid joint. 36 of these patients were reviewed and re-examined. We used the wrist-score as well as new X-rays for our retrospective examination. The untreated arthrosis of the scapho-trapezoid joint does not lead to any reduction of wrist movement and only in some cases to a slight decrease of grip strength (compared with the other side). 23 of the 36 patients were free of pain. Two thirds of the reviewed patients (24 of 36) did not feel any loss of usability of the hand. Only four in 36 cases suffered a significant loss of function. The X-rays for this follow-up showed an increase in arthrotic changes of the scapho-trapezoid joint in comparison to the X-rays after the resection-suspension arthroplasty of the carpometacarpal joint of the thumb in one-third of the patients. In conclusion, in the majority of the patients there are no clinical symptoms of an untreated scapho-trapezoid arthrosis after resection-suspension arthroplasty of the carpometacarpal joint of the thumb. Individual subjective symptoms did exist, even with increased radio-morphologic changes, only in a few cases. We cannot recommend combining of the resection suspension arthroplasty of the carpometacarpal joint of the thumb with the risky, uncomfortable fusion of the scaphoid and the trapezoid. Presently, we treat very severe arthrotic changes in both joints with a double-resection arthroplasty. These are very rare cases. If there are any advantages in this procedure, those will need to be proven in another follow-up study. PMID- 11917681 TI - [Experiences with Eaton-Littler reconstruction for therapy of ligament instability of the thumb saddle joint]. AB - Painful instability of the thumb carpometacarpal joint is a rare diagnosis. Between October 1995 and August 1999, eight operations of ligament reconstruction for unstable thumb carpometacarpal joints were performed. We report on the follow up of these patients which took place at an average time of 12.27 months (range 5 through 36 months) after the operation. Painful hypermobility of the non arthrotic thumb carpometacarpal joint was the indication for performing the operation in all cases. By this operation, the joint capsule is reinforced in two planes: the existing palmar ligament of the carpometacarpal joint is reinforced while it provides ligament support for the radial aspect of the joint that is otherwise membranous. In our study, we compared the pinch strength, the overall strength of the hand, the motion of the thumb, and radiographic findings for the amount of subluxation before and after performing the operation. In addition, each result was investigated with the DASH-questionnaire (DASH = Disability of Arm--Shoulder--Hand). The results were good or excellent in six cases, satisfactory in one case, and poor in one case. Pinch strength increased from a preoperative average of 19.6 N to 34.3 N and the vigor increased from 84.8 N to 113.1 N. Subluxation on the radiographs could be reduced from an average of 6.0 mm to 3.5 mm. All operated patients were investigated using the DASH questionnaire. Herein the average score was 22.92 on a scale whereby 0 reflects minimum and 100 maximum disability. CONCLUSIONS: Our experience with the Eaton Littler procedure for stabilisation of the hypermobile painful thumb carpometacarpal joint were mostly positive. The objective results as well as the results of the evaluation of the DASH-questionnaire could confirm this impression. PMID- 11917682 TI - [Clinico-epidemiologic aspects of primary pneumonia in the general population]. AB - During 2000 we made certain research into prevalence of the patients with an acute pneumonia at rural as well as town inhabitants of West Herzegovina (FBiH) within the population of 29,000 inhabitants 236 cases (0.81%) of acute pneumonia with specific clinical-lab results and the x-ray of the lungs, has been diaponesed in general population. Mortality was 2%. For the differentiation of bacterial from atypical pneumonia it was of very importance CRP (reactance of acute phase which was raised from the first day of pneumonia attack), the concentration of leucocytes with-in differential blood count. The average values of erythrocute sedimentation at the first checkup of the patients i.e. at the beginning of bacterial and atypical pneumonia were almost the same. But after a week the values were divergent and a little big higher at the patients suffered from bacterial pneumonia. PMID- 11917683 TI - [Central nervous system neoplasms in clinical data from the Neurology Clinic KCU in Sarajevo 1990-1999]. AB - The Neoplasms are the second leading cause of the mortality of the adult according to WHO. The last decade is the decade of the increase of all the epidemiologic parameters of the neoplasms in general, so also the neoplasms of the central nervous system. The goal of the work was to realize the frequency of the appearance of the neoplasms of the CNS on the clinical material of the Neurologic clinic CC University of Sarajevo with the special accent on the influence of the war as the specific stressor and the factor sui generis at the epidemiologic parameters of the neoplasms of the CNS. The study is retrospective: it is comprehended the period from the 1st January 1990 till 31st December 1999 year. The patients have been analyzed according to years, according the kind of the neoplastic processes and according to the time periods. At the analyzed period was treated 10,329 patients per year, according to the kind of the neoplastic processes and according to the time periods we evidenced the mild decrease from 1991-1999 year (3.03%-2.44%). According to the sex we register 167 males (60%) and 116 females (40%). The most frequent occurrence is in the age period from 55 to 69 years (55% patients). Of the primary neoplastic processes we had in 178 patients (64), with metastases 105 patients (36%). We analyzed the time periods: the prewar period (1st January 1990-5th April 1992), the war period (6th April 1992 till 14th February 1955), and the postwar period (15th December 1995 till 31st December 1999 year). The primary neoplasms in the postwar period we had in 70 patients (40%), in the course og est 35 patients (20%) and after war 69 patients (40%). The metastatic processes are in the evident increase during the war and after the war: before the war 19 patients (18%), during the war 32 patients (31%) and after the war 54 patients (51%). In the collected material during the war and after the war was evidenced the increase of the primary neoplastic processes at the lungs and other organs which have methastized at the nervous system. On the basis of our examination we can conclude that the total clinical material was present the mild percentage decrease of the neoplasms in the relation to all other admitted patients. The primary neoplastic process have been equally present in the patient material before and after the war with certain decrease during the war (probably because of the war and the reduced diagnostics). It is noticed the significant increase of the metastatic processes of the nervous system during the war and after the war. We are of opinion that the increase of the metastatic processes during the war and after the war is more many fold conditioned: the conditions of life of people during the war, shellings, fright from death and wounding, mourning for the killed and the wounded members of the most narrow family, the weak and inadequate nutrition, the increased smoking of the cigarettes, decrease of the total immune forces of an organism and others. PMID- 11917684 TI - [Treatment of Parkinson's syndrome]. AB - This paper discuss other possible approaches to the treatment od Parkinson's disease and parkinsonism, particularly in the case of younger patients. There is no doubt that levodopa treatment has the major advance in the clinical management of parkinsonism patients. However, the artificially of the levodopa decreases after several years and motor complications appear. Precise diagnosis and estimated degree of disease are used before any treatment of parkinsonism. Antiholineregigs or amantadine or dopamine agonists such as pergolide and bromcriptone are highly affective and they can delay the need for the levodopa. PMID- 11917685 TI - [Surgical treatment of urinary stress incontinence with the tension-free vaginal tape]. AB - TVT is the modern operative technique for the treatment of female urinary stress incontinence which was invented by Ulmsten and his co-operators. Tension-free vaginal tape implies the implantation of a prolene tape around mid-urethra via a minimal vaginal incision. It is the minimal invasive procedure under local anaesthesia and ambulatory performed. The majority of the women to be discharged from the clinic the same day or the day after the procedure. Results of studies about tension-free vaginal tape operation presents high success rate (90%) without serious complications, specially long term urinary incontinence. There is no signs of rejection the tape. PMID- 11917686 TI - [Rehabilitation of the shoulder joint after acromioplasty in the subacromial pain syndrome]. AB - Diagnostic error possibility and non-adequate surgical treatment by shoulder injury lead to non-adequate rehabilitation program. Authors show the case with diagnostic error and discuss the reasons for incomplete functional answer after rehabilitation-program. It is given the recommendation for solving these problems. PMID- 11917687 TI - [To whom does the journal "Medicinski Arhiv" belong]. PMID- 11917688 TI - [Myxoma of the right atrium and left ventricle]. AB - In this case report has been shown 63 years old woman who was admitted on internal medicine ward because of chest pain. She said, she has had chest pain for one year, exercise not dependent, connected with shortage of breath and often with coughing mixed with blood. One month before she was discharged from Neurologic Clinic where she had been admitted because of cerebrovascular insult. Echocardiographic analysis shows loose myxoma of right atrium on the thin stalk connected with interatrial septum with very probable myxomatosis in the areas of medial thirdness interventricular septum and inferior wall of left ventriculum, and persistence minor thrombus in acinetic apex of left ventriculum. It was pointed out vera rare location of myxoma and echocardiography as method od choice in its diagnoses. PMID- 11917689 TI - [Obesity as a risk factor for the onset of type 2 diabetes mellitus]. AB - Today overweight is the biggest public health problem in the world. According to statistical data, its occurrence is the most frequent among the English 53%, than German 51%, USA 48%, Italian and Spanish 40%, in French 38%, Swedish 30% and Russian 34%. According to available data 0.3% of the BiH population is overweight. Overweight can be related to collection of fat depos--obesity, or enlarged muscle mass--muscle hypertrophy. Criteria for evaluating of overweight are BMI (Body Mass Index). Obesity is the remarkable risk factor for the human health and it performs the background for many diseases--cardiovascular, diabetes mellitus, respiratory diseases, malignant diseases of gastrointestinal tract, joint-bone system diseases etc. For the regulation of optimal body weight, energy balance between input of energy by the food and its consumption is needed. PMID- 11917690 TI - [Internal contamination with depleted uranium and health disorders]. AB - In this review we used the published data on depleted uranium (experimental and epidemiological) from the current literature. Depleted uranium is a toxic heavy metal that in high dose may cause poisoning and health effects as those caused by lead, mercury, and chromium. It is slightly radioactive. The aim of this review was to select, to arrange, to present references of scientific papers, and to summarise the data in order to give a comprehensive image of the results of toxicological studies on depleted uranium that have been done on animals (including carcinogenic activity). We have also used epidemiological posted study results related to occupational and environmental exposure to depleted uranium. The toxicity of uranium has been studied extensively. The results of the studies indicated primarily its chemical toxicity, particularly renal effects, but depleted uranium is not radiological hazard. Uranium is not metal determined to be carcinogenic (the International Agency of Research on Cancer). The military use of depleted uranium will give additional insight into the toxicology of depleted uranium. The present controversy over the radiological and chemical toxicity of depleted uranium used in the Gulf War requests further experimental and clinical investigations of its effects on the biosphere and human beings. PMID- 11917691 TI - [Hexetidine--an oral antiseptic]. AB - Hexetidine is very safe oral antiseptic with broad antibacterial and antifungal activity in vivo and in vitro. It has local-anesthetics, astringent and deodorant activity. Also, it has very strong antiplac effects. Resistention of microorganisms on hexetidine is short and transient. These characteristics give important therapeutic role in treatment of oral infections. PMID- 11917693 TI - [Variations in the position and point of origin of the vermiform appendix]. AB - There were investigated variations of appendix vermiformis in the place of origin and position. The investigations were carried out on 50 human preparations of adults of both sexes, unintentional choice. The position and relation between intestinum caecum and appendix vermiformis were determined by forensic medical and pathoanatomical autopsy. Place of origin of appendix vermiformis, from wall of intestinum caecum is determined by anatomical dissection. Intestinum caecum has a variable shape and it occurs in two forms: conical, which dominates (56%) and square (44%). It has constant position in fossa iliaca dextra in 100% of 50 investigated cases. Appendix vermiformis is fully variable organ as for position and the place of origin from the wall of intestinum caecum. Dominantly (52%) it has rising position, and two subtypes are present: retrocecal (more expressed 38%) related to the retrocolic subtype (14%). Very frequent position of appendix vermiformis is a falling one (32%), in which pelvic position is more frequent (26%) related to descendent position (6%). Appendix vermifirmis is located subcaecaly in 8% of total number of investigated cases, found in three subtypes. It is found out that ostium appendicis vermiformis has a variable position in the wall of intestinum caecum. It is predominantly placed in the middle of the lower pole of the intestinum caecum (58%), in medial wall it is present in 32%, of all investigated, and in the lateral wall in the least number case 10%. The results of these investigations point out how important is to know variable anatomies of appendix vermiformis, for the clinical image of acute appendicitis is undoubtedly caused by the variable anatomical relations. PMID- 11917692 TI - [Czech and Polish women as the first female physicians in Bosnia-Herzegovina]. AB - Bajerova, Keckova, Krajevska and Olsevska--female physicians pioneers which worked in B&H. Born in Czech Republic and Poland, they began her medical work far from B&H, and full affirmation of her job received in this area. PMID- 11917694 TI - [Permeability of the hematoencephalic barrier by proteins in diabetes mellitus]. AB - Patient with diabetes mellitus is prone develop vascular lesion of blood vessels of all kinds. Microangiopathy and atherosclerosis are progressive during the illness. Following the study of Stanley I. Rapport the at all scientist on blood brain barrier permeability change in experimental conditions, also with this study blood brain barrier change in the diabetes mellitus patients was observed. In patients with diabetes mellitus the level total proteins increased in the cerebrospinal fluid. PMID- 11917695 TI - The geography of mental health. PMID- 11917696 TI - [Geographic analysis: epidemiologic studies and resource allocation]. PMID- 11917697 TI - The 'three waves' of research in mental health geography: a review and critical commentary. AB - OBJECTIVE: To consider research conducted in the sub-field of mental health geography, concentrating on work published in English. METHODS: The paper offers an comprehensive, in-depth and critical reading of the relevant literature on mental health geography since the inception of this subfield of inquiry in the early-1970s. RESULTS: The paper identifies three 'waves' of research within work on mental health geography. It describes these 'waves' in detail, interprets certain strengths and weaknesses of the first two 'waves', which are well established, and provides suggestions about important questions to be addressed in a future third 'wave'. CONCLUSION: Much excellent research has so far been undertaken within mental health geography, but there is scope to increase the relevance of this research through widening the focus of research and by being prepared to connect research more directly to mental health policy and politics. PMID- 11917698 TI - Progress in the psychotherapy of mood disorders: studies from the Western Psychiatric Institute and Clinic. AB - During the last three decades, we have witnessed dramatic improvements in both the psychosocial and pharmacological treatments of affective disorders. Administered in concert with the new medications advances in pharmacology have produced, disorder-specific psychosocial treatments have further improved the prognosis and course of bipolar and unipolar disorder. METHODS: We review our research on unipolar and bipolar disorder and their treatment, in particular interpersonal psychotherapy (IPT) and modifications thereof. RESULTS: We provide empirical evidence that IPT is an efficacious acute and maintenance treatment for affective disorders. Our cumulative research and clinical experience suggest that interpersonal relations and circadian and social rhythms influence affective illnesses and that psychotherapy may aid in normalizing problems in these areas for patients with affective illnesses. CONCLUSIONS: Despite the excitement generated by the recent progress in research on mental disorders and their treatment, we are yet to fulfill the promise that the explosion of knowledge about targeted pharmacotherapies or psychotherapies would seem to offer. To move our field forward, we must continue to apply scientific rigor and thought to understanding the suitability of current nomenclatures, the impact of comorbid psychiatric and medical illnesses and symptoms on the manifestation and treatment of affective disorders, and the practicality of widespread utilization of new treatments. PMID- 11917699 TI - Psychiatric genetics: recent advances and clinical implications. AB - OBJECTIVE: To present an overview on current progress and future directions in psychiatric genetics. METHODS: The review of studies that have demonstrated a genetic contribution to a wide range of psychopathology using family, twin, adoption studies and exploration of the methods and limitations of molecular genetic studies. RESULTS: Single gene disorders has been the area that is most straightforward with striking advances in disorders such as Huntington Disease and early onset familial Alzheimer disease. Complex phenotypes such as schizophrenia and affective disorder have presented greater difficulties but late onset Alzheimer disease and dyslexia are examples where replicated molecular genetic findings suggest that gene identification is feasible even for multifactorial disorders. CONCLUSION: The combination of increasingly complete information on the genome together with accessibility to this on the internet provide the essential tools for the search for susceptibility genes. Another essential requirement in trying to identify genes of small effect is well characterized large scale collections of cases and this demands the interaction of epidemiological and clinical researchers. Advances in genomics will also allow tailoring of pharmaceuticals pointing at treatment response and side effects. Hopefully all this perspectives together, will improve our understanding of the neurobiological pathogenesis of diseases such as Schizophrenia, Depression and Bipolar disorder 'legitimizing' them in the public view. PMID- 11917700 TI - [Psychiatric residential facilities in Italy. Results of phase I of the PROGRES project]. AB - OBJECTIVE: The 'PROGRES' (PROGetto RESidenze' Residential Project) project has 3 aims: 1. To survey all Italian psychiatric Non-Hospital Residential Facilities (NHRF) (Phase 1); 2. To assess in detail 20% of the NHRFs and the patients living there (Phase 2); and 3. To carry on training programmes for the mental health workers of these facilities (Phase 3). We report here the results of Phase 1. METHODS: All NHRFs were surveyed using a structured interview administered to the manager of the facility. In some cases, this information was supplemented with data gathered from other mental health workers at the NHRFs. RESULTS: On May 31, 2000, there were in Italy 1,370 NHRFs with 17,138 beds, giving an average number of beds per facility of 12.5 and a rate of 2.98 beds per 10,000 inhabitants. This rate varies greatly between regions, with a ratio of 1:10 between the two regions with the lowest and the highest bed rates. Seventy-three percent of the NHRFs have a 24-hour staff coverage; more than 50% are directly managed by the NHS Departments of Mental Health and more than three-quarters of the NHRFs are directly funded by the NHS. In the course of 1999 38% of the NHRFs discharged no patients, and another 32% discharged fewer than 3 patients. In about half of the NHRFs the most common patient age group was 40-59 years. In the Italian NHRFs there are 11,240 full-time mental health workers, plus several thousand half-time staff. The average number of full-time mental health workers in each NHRF is 8.2. In 58% of the NHRFs the operational chief is a psychiatrist; some 40% of the overall number of mental health workers in the NHRFs have no specific psychiatric training. The total number of patients resident in the NHRFs is 15,943; among them, 58% have never been admitted to a mental hospital, 40% have been admitted and a small percentage (1.6%) has been previously admitted to a forensic mental hospital. The availability of NHRF beds is negatively associated with the availability of non-hospital facilities (e.g., community mental health centres, day-centres) and positively associated with the number of beds in private psychiatric facilities in each region. DISCUSSION: The current rate of NHRF beds in higher than the officially recommended national standard (2/10,000 inhabitants). However, there is a great variability between regions, which is related to the overall provision of different types of psychiatric facilities. Most NHRFs provide intensive care, and the continuum of residential facilities with different types of care, support, degree of autonomy, etc. recommended by several authors for the long-term treatment of severe patients with different disabilities, seem to be lacking. NHRFs have a very low patient turnover rate, and this may create problems in the future. CONCLUSIONS: The PROGRES project is the first national study ever carried out in Italy to evaluate a specific type of psychiatric facilities in the context of the new system of psychiatric care. When all the data will be available, it will be possible to assess in detail for the first time a national representative sample of severe, long-stay patients and the care they are receiving; on this basis, it will also be possible to recommend evidence-based policies aimed at improving the care of the severely mentally ill. PMID- 11917701 TI - [Monitoring the impact on international literature of scientific production of Italian researchers, in the disciplines of "psychiatry" and "psychology." The period of 1995-1999 compared with the period of 1985-1989]. AB - AIM: The aim of this paper is to present the results of a citation analysis concerned with the impact on the international literature of Italian researchers in psychiatry and psychology and of sub-fields, within these two disciplines, in 1995-1999 compared to 1985-1989. METHOD: We used a database prepared by the Institute for Scientific Information (ISI) that included all papers, with at least one Italian author, working in Italy, which appeared in two periods (1995 1999 and 1985-1989) in psychiatric or psychological journals listed in Current Contents. A citation analysis concerning the following years, up to 1999, was performed. RESULTS: In 1995-1999 the most cited researchers were those involved in clinical psychology, psychiatry and psychopharmacology research. Neuropsychology, which dominated the period 1985-1989, underwent a clear-cut decline. CONCLUSIONS: Citation analysis, concerning relatively short periods of time (five years), appears to be a very helpful tool for monitoring the development of scientific research in specific disciplines and the trend of the scientific production of researchers. PMID- 11917702 TI - The Bill for Rights. PMID- 11917703 TI - Baby Grace. PMID- 11917704 TI - Brave new birds. The use of 'animal integrity' in animal ethics. AB - Suppose "chicken" eggs could be produced by quasi-chickens--genetically engineered humps of living chicken-flesh that do nothing but lay eggs. Would there be anything amiss with that? Animal ethicists invoke the notion of animal integrity in order to give intellectual content to the intuition that there would be. On inspection, 'integrity' isn't everything its proponents want it to be. Yet there's enough in it to make reasoned argument possible. PMID- 11917705 TI - Sex selection: not obviously wrong. AB - Although sex selection calls for careful thought, it seems in many cases to be neither intrinsically objectionable nor likely to have bad consequences. PMID- 11917706 TI - Sex selection in the United Kingdom. AB - The British have taken a comprehensive approach to regulating reproductive medicine. A loophole in the current law leaves some cases of sex selection uncovered; if that loophole were closed, however, the law is robust enough to address the concerns about sex selection while permitting it in many cases. PMID- 11917707 TI - Bioethics and the rituals of media. AB - Popular media may make short shrift of complex ideas and moral deliberations, but it can also serve bioethics well. Bioethics should embrace the ritual function of the media in bringing issues to public attention and in reinforcing bioethics as a field. PMID- 11917708 TI - Why physicians? Reflections on The Netherlands' new euthanasia law. PMID- 11917709 TI - Cloning and a right to procreate. PMID- 11917710 TI - Making room for alternatives. PMID- 11917711 TI - Death is the remedy? PMID- 11917712 TI - [Should magnetic resonance cholangiopancreatography replace endoscopic retrograde cholangiopancreatography in the diagnostic algorithm of biliary-pancreatic diseases?]. PMID- 11917713 TI - [Comparative study between magnetic resonance cholangiopancreatography and endoscopic retrograde cholangiopancreatography in the diagnosis of the pancreatic and biliary obstruction]. AB - BACKGROUND: The endoscopic retrograde cholangiopancreatography and magnetic resonance cholangiopancreatography were methods used for diagnosis of the diseases in the biliary and pancreatic ducts. AIM: Compare the diagnostic concordance among, analyzing sensitivity, specificity, positive predictive value, negative predictive value and accuracy. PATIENTS/METHODS: There were 41 patients in the studied group, divided in two groups: I--without biliary or pancreatic tract obstruction, and II--with obstruction. Group II was further divided in A- obstruction due to lithiasis, and B--due to other causes. RESULTS: Concordance between the two methods was found in 67% in group I and 82% in group II. Sensitivity of endoscopic retrograde cholangiopancreatography was 94% and magnetic resonance cholangiopancreatography was 89%. Specificity of endoscopic retrograde cholangiopancreatography was 100% and magnetic resonance cholangiopancreatography was 67%. Magnetic resonance cholangiopancreatography positive predictive value was 93%, the negative predictive value was 50% and the accuracy was 85%. CONCLUSION: Both methods showed the same sensitivity. PMID- 11917714 TI - [Intraesophageal pressure during esophageal speech in laryngectomized patients rehabilitated or no rehabilitated for oral communication]. AB - BACKGROUND: After laryngectomy for treatment of pharyngeal/laryngeal carcinomas the patients may be rehabilitated, for oral communication, with the esophageal speech. AIM: To study the intra-esophageal pressure during the esophageal speech. PATIENTS AND METHODS: It was measured the intra-esophageal pressure in 25 laryngectomized patients aged 40 to 70 years (median 57 years), 10 rehabilitated with esophageal speech and 15 unable to do so. The manometric method with continuous perfusion was used. The esophageal pressures was measured 3 to 5 cm below the upper esophageal sphincter when the patients tried to speak the vowel "a". Sometimes the air swallowed went to the stomach, with a peristaltic or simultaneous contraction in the esophageal body. RESULTS: During the attempt of esophageal speech the intra-esophageal pressure was higher in patients able to have esophageal speech (26.4 +/- 10.1 mm Hg, mean +/- SD) than in patients unable to do so (13.7 +/- 7.2 mm Hg). The esophageal contraction after a swallow of air was also higher in patients with esophageal speech (45.3 +/- 8.6 mm Hg) than in patients unable to do so (33.8 +/- 13.1 mm Hg). CONCLUSION: Laryngectomized patients rehabilitated with esophageal speech has a higher intra-esophageal pressure during speech than patients unable to do so, what may be consequence of the capacity to retain air inside the esophagus. PMID- 11917715 TI - [Contribution of hepatic immunoglobulin A deposits to the diagnosis of alcoholic hepatopathy]. AB - BACKGROUND: Alcoholic hepatic disease is a severe and frequent disease and its diagnosis is not always an easy task. AIM: To assess the contribution of immunoglobulin A (IgA) in the hepatic sinusoids for diagnosis of alcoholic hepatopathy. PATIENTS AND METHODS: The presence of IgA was studied through direct immunofluorescence in 59 patients submitted to hepatic needle biopsy, indicated by clinical or in vitro changes suggestive of chronic hepatopathy. RESULTS: A significant deposition of IgA was found in alcoholic patients as compared to non alcoholic patients, with 76% sensitivity (95% CI: 54.5-89.8) and 73.5% specificity (95% CI: 55.3-86.5). In individuals who present only alcohol as the etiological agent of hepatopathy, compared with the subgroup of B or C virus carriers, the results were even more significant, with 85.7% sensitivity (95% CI: 56.2-97.5) and 89.5% specificity (95% CI: 65.5-98.2). CONCLUSION: The deposition of IgA in the hepatic sinusoids present sensitivity and specificity for the diagnosis of an alcohol-induced hepatic lesion. This resource can be particularly useful when conventional histology can not be define a specific cause for the change found. PMID- 11917716 TI - [Return to work of adults after liver transplantation]. AB - BACKGROUND: Orthotopic liver transplantation is an established therapy for patients with end-stage liver disease. In the last years more attention has been given for the improvement of the quality of life after liver transplantation, and the return to important life pursuits, such as employment, are important goals of this type of therapy. AIM: To assess the work's return rate in liver transplant recipients and the reasons for inability after liver transplant. PATIENTS AND METHODS: Forty-one adults (age > or = 18 years) who underwent liver transplantation at our institution, between September 1991 and June 1999, with a post-transplant survival > or = 9 months, good graft function and a regular clinical follow-up were enrolled for the study. A questionnaire that measured aspects of inactivity before and after liver transplantation had been applied. RESULTS: Thirty-one of 41 patients were unable to work due end-stage liver disease before liver transplantation. Return to work was observed in 28 of 41 patients (68%) in a mean time of 5.9 months after the surgical procedure. Eleven of 28 active patients after liver transplant (39%) contribute significantly to family income. The reasons for inactivity after liver transplantation were: early retirement (n = 5), unemployment (n = 4), and physical disability (n = 4). CONCLUSION: Successful liver transplantation not only improves the survival rate but allows patients to return to social function and workforce. PMID- 11917717 TI - [Assessment of gliadin in pharmaceutical products - important information to the orientation of celiac disease patients]. AB - BACKGROUND: Some drugs might contain gliadin which can be dangerous for celiac disease patients. OBJECTIVE: Detect gliadin in pharmaceutical products commonly used in Brazil. METHODS: We analyzed 78 pharmaceutical products selected aleatory from a list of 180 products most frequently sold at Brazilian community pharmacies. The analyzed samples were analgesics (n = 9), anthelmintics (n = 3), antacids (n = 8), antibiotics (n = 13), anticholesteremics (n = 1), anticonvulsants (n = 2), antidepressants (n = 2), antiemetics (n = 3), antihypertensives (n = 3), antihistaminics (n = 3), anti-inflammatories (n = 7), antipyretics (n = 2), bronchodilators (n = 1), laxatives (n = 1), oral contraceptives (n = 5) and vitamins (n = 10). The samples were analyzed by enzyme immunoassay based on monoclonal antibodies omega-gliadins, the elected technique according to the Codex Alimentarius Commission WHO/FAO. All samples were analyzed in duplicate. The sensitivity of this test is 4 mg of gliadin/100 g of product. RESULTS: Only one (1.3%) out of 78 pharmaceutical products contained detectable amounts of gliadin (5.5 mg/100 g). The active ingredient of this drug is ranitidine. According to the Codex Alimentarius Commission WHO/FAO the intake of 10 mg of gliadin/day should not be exceeded by celiac disease patients. Considering the amount of gliadin in each capsule of ranitidine, the ingested quantity would be lower than the maximum allowed for celiac patients. CONCLUSIONS: In this study gliadin was not detected in pharmaceutical products in harmful amount for celiac disease patients. PMID- 11917718 TI - [Study of renal glucose release in rabbits submitted to total functional hepatectomy and norepinephrine infusion]. AB - AIM: To study the possible endogenous sources of glucose in the absence of the liver (equivalent to the anhepatic period of liver transplantation). MATERIAL AND METHODS: A experimental model of total functional hepatectomy in anesthetized rabbits was developed. The aorta and the right renal vein were catheterised in order to collect blood samples to measure glucose contents. The animals were divided into two groups: group 1, 5 animals underwent only norepinephrine infusion; group 2, 15 animals underwent norepinephrine infusion and submitted to total functional hepatectomy. RESULTS: In group 2, before the hepatectomy, arterial glucose levels were higher than venous ones and after the liver removal, the venous levels became higher than the arterial ones. This pattern showed an inversion in the glycemic curves. In group 1 this pattern was not observed. CONCLUSION: The glycemic curves behavior observed in group 2 its not due to norepinephrine infusion, but represents renal glucose release after total functional hepatectomy. PMID- 11917719 TI - [Electromanometry of the upper esophageal sphincter before and after esophageal perfusion with hydrochloric acid 0,1N. Experimental study in dog]. AB - BACKGROUND: The responses of the upper esophageal sphincter to gastroesophageal reflux is controversial. OBJECTIVE: Study the effect of upper esophageal sphincter to the esophageal acid perfusion. METHODS: Thirty adult dogs of both sexes were studied, being submitted to esophageal electromanometry. The pull through technique and continuous infusion of the catheters with distilled water were employed. These exams allowed us to measure the pressure width (mm Hg) and the length (cm) of the upper esophageal sphincter in basal conditions (moment 1). After this first phase the animals were submitted to esophagic infusion, being then divided in three groups, according to the solution used in the infusion and the moment of the study, as follows: Group 1: esophagic infusion with distilled water and electromanometric studies accomplished 15 minutes (moment 2) and 30 minutes (moment 3) of the end of the infusion. Group 2: esophagic infusion with HCl 0.1 N and electromanometric studies accomplished 15 minutes after the end of the infusion (moment 2). Group 3: esophagic infusion with HCl 0.1 N and electromanometric studies accomplished 30 minutes after the end of the infusion (moment 3). RESULTS/CONCLUSIONS: This research was performed to evaluate the esophagic acidification influence on the upper esophageal sphincter. The observed results allowed us to conclude that the acidification of the esophagus did not cause any alteration on the pressure width and on the length of the upper esophageal sphincter. PMID- 11917720 TI - [Hereditary hemochromatosis. Case report and review of the literature]. AB - BACKGROUND: HFE-associated hemochromatosis is one of the most common inherited liver disease in Caucasian populations and refers to the association of increased iron stores with tissue damage (e.g., cirrhosis, diabetes, cardiomyopathy), which is progressive when diagnosis and treatment are delayed. AIMS: Description of a case of hereditary hemochromatosis in an asymptomatic 44-years-old patient, whose diagnosis was made through casual identification of abnormal iron markers. We will also present a brief review of the literature about the topic. CONCLUSIONS: Hereditary hemochromatosis is an ideal disease for primary prevention since the disease can be detected well before serious complications develop. This intervention has been shown to prevent the manifestations of the disease, altering its natural history. PMID- 11917723 TI - [A few practical ideas concerning the application of the law on voluntary pregnancy interruption]. AB - The law on voluntary pregnancy termination has been modified in France. Many questions are currently raised concerning its application. We provide a review discussing available elements needed to answer these questions. PMID- 11917721 TI - Triple therapy with clarithromycin, amoxicillin and omeprazole for Helicobacter pylori eradication in children and adolescents. AB - BACKGROUND: Helicobacter pylori infection presents high prevalence in developing countries, but there are few pediatric assays evaluating antimicrobial treatment. OBJECTIVE: The aim of this study was to investigate Helicobacter pylori eradication rate using a short regimen (7 and 10 days) of triple therapy with clarithromycin, amoxicillin and omeprazole. PATIENTS AND METHODS: Twenty-five Hp positive patients who presented severe epigastralgia, were submitted to antimicrobial treatment with amoxicillin (50 mg/kg/day--maximum dose 1 g bid), clarithromycin (30 mg/kg/day--maximum dose 500 mg bid) and omeprazole (0.6 mg/kg/day--maximum dose 20 mg bid) during 7 or 10 days. After 2 months, clinical symptoms were evaluated and gastric biopsies were taken to test Hp eradication. RESULTS: Overall eradication rate was achieved in 16/25 patients (64%--IC(95% = 45-83%), in 11/15 (73%--IC(95%) = 51-95%) patients who used 10 days therapy course and in 5/10 (50%--IC(95%) = 19-81%) who used 7 days therapy course. Eradication drugs were well accepted and adverse effects were reported in two patients (8%). CONCLUSIONS: This triple therapy regimen had moderate efficacy (64%). The data suggests that 10 days therapy course achieves better eradication rate (73%) than 7 days course (50%) to treat Hp infection in our population. PMID- 11917722 TI - [New drugs for the treatment of functional dyspepsia]. AB - BACKGROUND: Functional dyspepsia is defined by upper gastrointestinal symptoms without any evidence of structural abnormalities or organic disease. Current pharmacological treatment of functional dyspepsia is largely empirical and involves anti-secretory or prokinetic drugs. AIMS: To review recent advances in the understanding of the mechanisms involved in symptom production in functional dyspepsia, as well as the development of new drugs that may interfere with these mechanisms, which may lead to more rational and effective treatment of this clinical condition. METHOD: Systematic review of papers published in English for the last 10 years. RESULTS: New drugs that increase propulsive gastroduodenal motor activity include new benzamides similar to cisapride, CCK-A blockers, agonists of opiate receptors and motilin agonists similar to erythromycin. A number of agents, including sumatriptan and buspirone, stimulates serotonin receptors in the myoenteric plexuses and have been shown to increase gastric accommodation to a meal. Finally, a number of new drugs that either increase thresholds for visceral perception or modify sensations is currently under investigation. This includes agents of several groups, such as octreotide, loxiglumide, ondansetron and other serotonin blockers, fedotozine and tricyclic antidepressant at low doses. CONCLUSIONS: Although these new drugs may improve the pharmacological approach to the treatment of functional dyspepsia, there is a need for randomized, controlled trials to assess their efficacy. Moreover, difficulties related to the identification of the mechanisms underlying symptoms may limit the utilization of these new drugs. PMID- 11917724 TI - [Clinical evaluation of urinary incontinence in the woman]. AB - Clinical assessment of stress and urge incontinence in women is always necessary before making therapeutic decisions. During the full bladder examination stress incontinence triggered by cough may suggest cervicourethral hypermobility. Leakage during the Vasalva manoeuvre may reveal incontinence due to intrinsic sphincter deficiency. The pad test provides an objective quantitative measurement of the degree of incontinence. Symptom scores enable intra- and inter-individual comparisons. Psychosocial implications are studied with specific quality-of-life scales. Together, these tests improved therapeutic strategies for managing stress and urge incontinence, providing an objective evaluation of the different treatments and thus a better medico-economic approach to the problem. PMID- 11917725 TI - [Contribution of diagnostic ultrasonography in cases of repeated embryonic implantation failure in fertilization in vitro]. AB - INTRODUCTION: To assess ultrasonic data in the context of repeated failure of implantation in assisted reproduction medicine. MATERIALS AND METHODS: We reviewed ultrasonic data (uterine score at follicular and luteal phase). The same ultrasound exploration was repeated for two consecutive cycles to assess the changes in abnormalities observed and compare findings in 16 patients with repeated failure of implantation and in 14 controls. The controls were women who became pregnant within the three months following the exploration. We then introduced acetylsalicylic acid and prednisolone as first line treatment and nitroglycerin as second line treatment to measure the effects induced in 14 pathological uterine scores. RESULTS: The score obtained under hormonal treatment was representative of other cycles (r = 0.812, p < 0.0014). The score was significantly lower and the luteal uterine artery pulsatility index was significantly higher in the repeated failure group than in the control group (p < 0.0001 and p = 0.008 respectively). For the 14 patients, treatment improved the uterine score in 8 with acetylsalicylic acid and prednisolone and in 4 with nitroglycerin. Two patients did not respond to treatment. Concerning the effect of therapy, of 14 patients, 8 had an improved uterine score with A + P, and with T; 2 patients did not respond to any treatment. CONCLUSION: The uterine score and luteal artery pulsatility index may be valuable tools for patients with implantation failure who all do not respond to the same therapy. PMID- 11917726 TI - [Contribution of pelvic floor MRI in the morphological and functional analysis of pre and postoperative levator muscle in patients with genital prolapse]. AB - OBJECTIVES: This study was directed to evaluate the morphological and dynamic aspects of the pelvic floor muscles by the MRI in patients with gynecological prolapse before and after surgery. MATERIALS AND METHOD: MRI had been made before surgical repair in 13 patients with gynecological prolapse, in 9 of this group MRI had been made after the surgery and the group of control was formed by 4 healthy women. We had used morphologic sequences T2 (TSE) and T1 (SE) and fast sequences T2 (TSE) in different positions: at rest, straining and retention for describe and evaluate the anatomic modification of the levator ani and the changes observed after surgery. RESULTS: The MRI is able to identify the changes of the levator ani muscles: intrinsic degeneratives lesions, increase of the muscular laxity especially for the puborectalis muscle and the wider of the levator hiatus. After surgical repair, the levator hiatus width and the laxity of the puborectalis muscle are slightly modified during straining. CONCLUSION: There are lesions in the muscular structure of the pelvic floor in patients with gynecological prolapse. The MRI was able to analyse these lesions very well especially after the use of fast sequences. The MRI is the future for exhaustive and non invasive study of the static pelvic disorders. PMID- 11917728 TI - [Maternal mortality in France: frequency, trends and causes]. AB - Ten years ago the first epidemiological surveys on maternal mortality in France were carried out on the national level. In 1995, a National Committee of Medical Experts was created to conduct confidential inquiries into maternal deaths. It is thus useful to examine the general picture of maternal mortality in France drawn by the routinely and permanently collected data. These statistics are collected independently of the procedure adopted by the National Committee on confidential inquiries into maternal deaths. National death and cause-of-death registries have recorded maternal death rates for several years with data by age, area of residence, nationality, and direct or indirect obstetric causes. The low and underestimated rate of 8.5 maternal deaths per 100,000 live births recorded in 1989 increased regularly up to 1992. Currently the rates have been around 9 to 13 with no evidence of a declining trend. The larger urban area around Paris (Ile-de France) has shown a statistically significant higher rate over the last several years. Post-partum hemorrhage remains the leading cause of maternal death. Compared with other European countries, maternal mortality in France is in an average position, similar to Great Britain (12 per 100,000), but higher than in Scandinavian countries. The elevated mean age of mothers at delivery is one explanation for the lack of a decline in the rate of maternal deaths expected until 2005 although further actions should be implemented to attempt to lower the rate to that observed in Scandinavian countries. A pertinent classification of causes of maternal deaths allowing valid international comparisons would be useful for helping answer the questions raised by clinicians. PMID- 11917727 TI - [Winter supplementation in the 3rd trimester of pregnancy by a dose of 80,000 IU of vitamin D]. AB - A non-comparative study was conducted to examine the effects of 80,000 IU vitamin D given in a single dose to 59 pregnant women from northern or southern France between their 27th and 32nd week of gestation during the winter season. Serum levels of 25 hydroxy-vitamin D (25 OH D), intact Parathyroid Hormone (iPTH), calcium, phosphates, proteins were measured at the inclusion, at delivery (mother and arterial cord) and in the newborn between the 3rd and the 5th day of life. The mothers' sun exposure and their vitamin D dietary intakes were evaluated with scores at the inclusion and at delivery. Before vitamin D supplementation, 34% of the women had a 25 OH D concentration below 10 ng/ml and 32% had hypocalcemia. At delivery, only one woman had a low 25 OH D concentration, whereas 15% of the women showed hypocalcemia. No neonatal hypocalcemia was observed and no vitamin D overdose was recorded in this study. The mothers' vitamin D dietary intakes were quite high; the lack of sun exposure during last summer appeared as a major vitamin D deficiency risk. A single dose of 80,000 IU vitamin D, taken between the 27th and the 32nd amenorrhoea weeks in winter, seems to be a good compromise between efficacy and tolerance. PMID- 11917729 TI - [Occupational toxic exposure in the pregnant woman. 1: principles fo individual risk assessment ]. AB - CONTEXT: Many women of childbearing age are occupationally exposed to chemicals and concerned with the ensuing risk when pregnant. OBJECTIVES: To describe the principles of individual risk assessment to be applied in pregnant women or women wishing to become pregnant that are exposed to chemicals at the workplace. METHOD: Each request for risk assessment is based on a comprehensive review of the hazards of the handled products together with a thorough evaluation of the actual exposure at the workplace. A toxicological advice is then written to the gynecologist or the general practitioner in charge of the patient. When the exposure is estimated to be hazardous for the pregnancy, either total withdrawal, avoidance of certain activities or improvements of individual protective devices are recommended. The outcome of the pregnancy is systematically followed-up. CONCLUSION: An objective assessment of toxic risks in pregnant women exposed to chemicals at the workplace can be done. Thus, patients who must be withdrawn or benefit from improvements of their workstation can be selected. PMID- 11917730 TI - [Occupational toxic exposure and the pregnant woman. 2: results of a prospective study of 100 pregnancies]. AB - CONTEXT: Many women of childbearing age are occupationally exposed to chemicals and concerned with the ensuing risk when pregnant. OBJECTIVES: To present the results of a prospective follow-up study of 100 pregnant women and to discuss them after a brief overview of the published data on this topic. METHODS: Since January 1996 the Lyon Poison Center has been conducting a prospective follow-up of all request concerning pregnant women occupationally exposed to chemicals. A thorough evaluation of the hazards of the handled products and of the actual exposure at the workplace is done for each patient. A toxicological advice is given and the outcome of the pregnancy is followed-up. RESULTS: One hundred pregnant women were included between January 1996 and December 2000. Based on the nature of the handled products, two groups have been identified: the first included 73 women exposed to organic solvents and the second 27 women exposed to miscellaneous. When the exposure was considered potentially hazardous for the pregnancy, either withdrawal from the workstation (19 cases), avoidance of certain activities (9 cases) or improvement of individual protective measures (29 cases) was recommended. In 43% of the cases, the occupational exposure was not considered hazardous to the outcome of the pregnancy. No increase of adverse outcome was identified: 4 miscarriages and 96 living births were observed, with 2 major malformations and 1 minor malformation. CONCLUSION: Occupational exposure to chemicals was not found to affect adversely the outcome of these 100 pregnancies. PMID- 11917731 TI - [Nasal endometriosis: apropos of 1 case]. AB - The nasal mucosa is an exceptional localization for endometriosis. We report a case observed in a woman who developed nasal tumefaction associated with epistaxis that had followed a cyclic repetitive pattern since puberty. Surgical excision of the nasal nodule confirmed the diagnosis of endometriosis. Local and general evaluated identified other localizations. Outcome has been favorable at short and mid term. Endometriosis is a frequent condition although extra-pelvic localizations are uncommon. The nasal localization is highly exceptional. PMID- 11917732 TI - [Placental teratoma: differential diagnosis with fetal acardia]. AB - The acardiac fetus is a rare congenital malformation observed only in monozygotic twin pregnancies (1/35.000 pregnancies). Placental teratoma is extremely rare (less than 20 cases have been reported in the literature since 1925). A case of placental teratoma, misdiagnosed as fetus acardius amorphus, is presented with ultra-sound follow-up, delivery and outcome of the normal fetus. Pathological findings, histogenesis, differential diagnosis with the acardiac fetus and finally review of the literature will be discussed. PMID- 11917733 TI - Advances in fetal and neonatal surgery for gastrointestinal anomalies and disease. AB - The last decade has seen considerable improvement in the understanding and treatment of neonatal surgical disorders. Translation of basic molecular biology research to clinical practice has directly improved the understanding and treatment of a number of congenital, developmental disorders, such as Hirschsprung's disease and congenital hyperinsulinism. Miniaturized instruments and improved optics have permitted increased use of videoscopic and minimally invasive techniques to even the smallest infants. Continued improvements in prenatal imaging will permit enhanced understanding of the prenatal natural history of congenital structural disorders and the development of more specific therapies. Finally, rigorous clinical research tools have begun to be applied to rare pediatric surgical disorders with the use of organized multicenter trials. It is an exciting time for all involved in the care of neonates. PMID- 11917735 TI - Gastroesophageal reflux in the neonate. AB - Physiologic GER may be a maturational phenomenon, because infants outgrow this over time. Many aspects of GERD in neonates and young infants remain incompletely understood, however, particularly the pathophysiology and long-term problems in high-risk neonates. Diagnostic and therapeutic availability is vital in infants with GER and airway compromise, apnea events, or chronic lung disease. Although most infants improve with conservative therapy, there is a dire lack of ideal pharmacologic agents that work on all the mechanisms of GER with the least consequences. Studies that permit diagnosis not only of the disease but also of the causal mechanism, better feeding strategies, and prevention of morbidity from GERD will be beneficial. PMID- 11917734 TI - Pharmacology for the gastrointestinal tract. AB - A variety of drugs are used in the neonatal nursery for the management of feeding intolerance, gastroesophageal reflux, and acid-related disease. Although the pharmacokinetics of some of these drugs have been described in infants and older children, further data are needed, particularly for preterm infants. No data are available characterizing the disposition of the proton pump inhibitors, which will likely be used in infants with refractory, acid-related disease. Further data are also needed to characterize fully the pharmacodynamics, or efficacy, of many of the commonly used drugs. PMID- 11917736 TI - Update on the etiologies and management of neonatal cholestasis. AB - The early detection of cholestatic liver disease is one of the major challenges facing pediatricians when evaluating the jaundiced infant. Early recognition of liver disease greatly facilitates the care and outcome of infants, because several serious life-threatening disorders may have cholestasis as a major presenting sign of underlying neonatal liver disease. A key component of the work up is measurement of serum conjugated bilirubin levels, which if elevated should prompt the clinician to initiate a work-up to determine the cause of neonatal cholestasis. In general, if a patient is developing progressive jaundice soon after birth, is still jaundiced at 2 weeks of life, or develops jaundice within the first month of life, a work-up for neonatal cholestasis should begin. A number of previously undiagnosed causes of neonatal cholestasis are beginning to be assigned genetic and infectious etiologies, with significant implications for the work-up and management of cholestatic infants. PMID- 11917737 TI - Update on management and treatment of short gut. AB - Management of patients with short-bowel syndrome represents a formidable challenge. Aggressive treatment including nutritional care and anticipation of potential complications and rapid treatment of complications enhance outcome. New therapies offer the promise of significantly improving morbidity and mortality. Intestinal transplant is appropriate for infants who would otherwise die from liver disease, recurrent sepsis, or lack of venous access. PMID- 11917738 TI - Newborn intestinal circulation. Physiology and pathophysiology. AB - The physiologic characteristics of the newborn intestinal circulation are unique when compared with the adult condition. Most important, intestinal vascular resistance across newborn intestine is exceptionally low and this transient reduction is mediated by an increased constitutive and stimulated production of NO. The low vascular resistance characteristic of newborn intestine alters the capacity of this vasculature to respond to systemic circulatory perturbations, such as hypotension and arterial hypoxemia. The essential role of endothelial production of NO in maintaining newborn intestinal hemodynamics might be important in the pathogenesis of NEC, because endothelial dysfunction would limit, or possibly eliminate, NO production, leading to substantial intestinal ischemia. PMID- 11917739 TI - Update on host defense and immunonutrients. AB - Neonatal intensive care in the past three decades has provided exciting modalities for improving the survival of critically ill neonates. There remains a great need for improving the quality of life for these survivors. In this article, the role the developing GI tract and its microenvironment play in the well-being of the neonate has been emphasized. Future therapies based on manipulation of the GI tract and its microenvironment by functional foods, immunonutrients, or pharmacologic agents may have effects not only during the neonatal period, but also throughout the individual's entire life. PMID- 11917740 TI - Key nutrients and growth factors for the neonatal gastrointestinal tract. AB - The nutritional support of gastrointestinal growth and function is an important consideration in the clinical care of neonatal infants. In most health infants, the provision of either breast milk or formula seems to support normal intestinal mucosal growth, but the most significant advantages of breast milk may be for host defense or gut barrier-related functions that are involved in reducing infection. The specific effects of various milk-borne growth factors on key mucosal immune and barrier functions are likely to provide valuable new clues to the advantages of human milk. A substantial number of preterm, low-birth weight babies or those suffering from compromised intestinal function, however, often cannot tolerate oral feedings and instead receive TPN. The consequences of TPN on gastrointestinal function and how this contributes to morbidity of these infants warrants further study, with respect to both clinical and basic research questions. Although enteral nutrition seems to be a critical stimulus for intestinal function, the minimal amounts and composition of nutrients necessary to maintain specific intestinal functions remain to be established. The experimental tools exist to start defining the specific nutrient requirements for the infant gut and some of these nutrients are known (e.g., glutamate, glutamine, and threonine). Peptide growth factors and gut hormones clearly play a role in gut growth and in several ways mediate the trophic actions of enteral nutrition. Although a number of these growth factors are good candidates for therapeutic use, their clinical application in the management of gastrointestinal insufficiency and disease has been slow. The emergence of GLP-2 as a trophic peptide that seems to target the gut is a promising candidate on the horizon. PMID- 11917741 TI - Development of the enteric nervous system. AB - Development of the ENS requires the function of a diverse set of genes encoding transcription factors, signaling molecules, and their receptors. Mutations of these genes result in altered ENS function in animals and humans. In particular, such mutations have been shown to contribute to many cases of Hirschsprung's disease. Elucidation of the mechanisms of ENS development and function allow the development of new approaches to the diagnosis, therapy, and prevention of human disorders of gastrointestinal motility. PMID- 11917742 TI - The controversy over screening mammography. PMID- 11917743 TI - Images in medicine. "Dropped" gallstones. PMID- 11917744 TI - Achieving universal health care coverage in Rhode Island: where are the challenges? PMID- 11917745 TI - Smallpox: clinical and epidemiologic features. PMID- 11917746 TI - Sleep disorders and a physician's responsibility. PMID- 11917747 TI - "No autopsy--he's suffered enough.". PMID- 11917748 TI - Sleep disorders: common but often unrecognized medical problems. PMID- 11917749 TI - Sleep, fatigue, and medical training: an overview. PMID- 11917750 TI - Approach to the sleepy patient. PMID- 11917751 TI - Behavioral treatment for chronic insomnia. PMID- 11917752 TI - A review of the adult primary sleep parasomnias. PMID- 11917753 TI - Therapy of insomnia. PMID- 11917754 TI - Web medicine. PMID- 11917756 TI - How to achieve accurate spirometric measurements. PMID- 11917755 TI - Occupational asthma: essential to recognize, easy to miss. PMID- 11917757 TI - Cytokine-modifying therapies in immune disorders. PMID- 11917758 TI - The differential diagnosis and workup of pruritus. PMID- 11917759 TI - Occupational medicine: physician assistants can meet the need. PMID- 11917760 TI - Let's get serious about promoting PAs. PMID- 11917761 TI - A realistic take on tech. PMID- 11917762 TI - The benefits of bar coding. PMID- 11917763 TI - Hospital ratings here to stay. PMID- 11917764 TI - Serving the uninsured. PMID- 11917765 TI - Defined contribution. PCAs (personal care accounts) coming soon? PMID- 11917766 TI - Charity care. 'Not just a plaque'. PMID- 11917767 TI - Clinical technology. Getting rid of the hassles. PMID- 11917768 TI - Quality of care. Ratings could pay. PMID- 11917769 TI - The medical workplace. No abuse zone. PMID- 11917770 TI - Practical. Throw out the squeeky wheel? PMID- 11917771 TI - Emergency diversions. ED teamwork. PMID- 11917772 TI - 1Q[3a]. What will be the field's top ethical issue in five years? PMID- 11917773 TI - HIMSS report. No job vacancy. PMID- 11917774 TI - HIMSS report. CIOs: ready to lead? PMID- 11917775 TI - Procurement. Groceries or meds? PMID- 11917776 TI - Rule14. Leadership means more than taking charge. It means doing what's right. AB - Leadership is difficult to define, but retired Gen. Norman Schwarzkopf says it comes down to two essential rules: when placed in charge, take command; and do what's right. PMID- 11917777 TI - A multitude of strains add up to one big headache. Feeling the pressure? AB - A multitude of strains--most beyond the control of health care executives--are converging to create major headaches for hospitals and systems. Skyrocketing insurance premiums, surging drug spending, Medicaid cuts, a staffing crisis and huge hikes in liability insurance rates contribute to the unprecedented confluence of pressures. And there's more. PMID- 11917778 TI - Building boom. AB - After a period in which financial issues forced many hospitals to postpone needed construction projects, a building boom is under way at last. Aging facilities, many dating to the heyday of Hill-Burton funding after World War II, are ill equipped for the enormous changes in health care delivery, from new technology to the emphasis on outpatient treatment. Innovative financing strategies are helping to feed the trend. PMID- 11917779 TI - An investment in community. AB - Memorial Hospital and Health System of South Bend, Ind., is this year's Foster McGaw Prize winner. Since 1993, Memorial has invested $20 million in nearly 100 community programs, including a kids' health museum, a homeless center and senior activities. PMID- 11917780 TI - Charting health care's future. AB - This month's fold-out section compiles data illustrating the impact that aging baby boomers will have on health care in the coming years. The data is taken from the Digest of Health Care's Future, H&HN's 54-page chartbook of statistics and analysis. PMID- 11917781 TI - Promises to keep. American hospitals have a pact with patients that must be kept. PMID- 11917782 TI - Scabies and lice: review of the clinical features and management principles. AB - Health care providers frequently encounter human infestations of scabies and lice. When a person is identified as having either of these infestations, he/she should be treated. More importantly, their close contacts should be examined and treated if needed. It is vital for dermatology nurses to understand the epidemiology, transmission, symptoms, distribution, diagnosis, complications, and treatment for scabies and lice infestations. PMID- 11917783 TI - Hair loss: an overview. AB - Hair loss is a common problem in men and women. Correct diagnosis of hair disorders is complex and requires evaluation of clinical presentation, history, physical examination, and laboratory tests. Hair loss may be categorized as hair shaft abnormalities, permanent alopecia, or nonpermanent alopecia. Nonpermanent alopecia, the most common type, includes androgenetic alopecia, telogen effluvium, alopecia areata, and traction alopecia. The hallmark of this group is the possibility of complete regrowth with adequate treatment. PMID- 11917784 TI - The sun's damaging effects. AB - Many people find exposure to the sunlight enjoyable and relaxing. In fact, most people agree that a tan looks great. For years, sunlight has been viewed as beneficial to one's well-being both emotionally and physically (Ness, Frankel, Gunnell, & Smith, 1999). However, exposure to ultraviolet radiation has its drawbacks. Sunlight is responsible for wrinkling, blotching, drying, and leathering of skin. In fact, 90% of all skin cancers result from long-term exposure to ultraviolet radiation. It is important to be educated about the effects of ultraviolet radiation and learn to balance its beneficial effects with its negative effects. PMID- 11917785 TI - What's your assessment? Psoriasis with steroid atrophy. AB - The "What's Your Assessment?" series includes a short case presentation and differential diagnosis. It is followed by a discussion of the disease or condition and the rationale used in each step of the assessment. PMID- 11917786 TI - Wound assessment and evaluation. AB - The "Wound Assessment and Evaluation" series includes a short case presentation and differential diagnosis. It is followed by a discussion of the disease or condition and the rationale used in each step of the assessment. PMID- 11917787 TI - What are those white patches on my patient's skin? AB - Vitiligo is a skin disease that affects 1% of the population. Vitiliginous skin does not contain pigment-producing cells called melanocytes. This leaves the appearance of white patches on the skin, which can either be generalized or localized. It is important for dermatology nurses to understand the basic pathophysiology of vitiligo, the various treatment methods, how to protect vitiliginous skin, and the psychological impact of the disease. PMID- 11917788 TI - Scabies. AB - The "Clinical Snapshot" series provides a concise examination of a clinical presentation including history, treatment, patient education, and nursing measures. Using the format here, you are invited to submit your "Clinical Snapshot" to Dermatology Nursing. PMID- 11917789 TI - Tattooing a major route of hepatitis C infection. PMID- 11917790 TI - Environmental dangers pose a threat to children's skin. PMID- 11917791 TI - Most Americans are not concerned with contracting genital herpes: survey. PMID- 11917792 TI - [Macular translocation. Improved prognosis for age-related macular degeneration]. PMID- 11917793 TI - [Visual prognosis and patient selection]. AB - Choroidal neovascularization (CNV) is one of the leading causes of central vision loss. Laser photocoagulation and photodynamic therapy may reduce the risk of severe loss of visual acuity in subfoveal CNV secondary to age-related macular degeneration. However, the most favorable effect of these therapies is the slowing of the vision loss. Macular translocation surgery, removing the neurosensory retina away from the area of choroidal neovascularization to a healthier underlying tissue, may improve or stabilize the visual acuity. Several pilot studies of this surgery reported visual recovery in selected patients. Well designed, controlled randomized studies are needed to clarify the role of the macular translocation surgery in treatment of choroidal neovascularization. Given the severe complications of this surgery, it is of utmost importance to discuss the potential risks and advantages of this procedure with the patient. PMID- 11917794 TI - [Surgical procedures in the treatment of age-related macular degeneration]. AB - The surgical techniques for removal of a subretinal membrane associated with age related macular degeneration will be discussed and compared on the basis of published data. The so-called simple subretinal membrane extraction will be compared with pigment epithelium cell transplantation and with the various techniques for macular translocation. The pars plana gas injection with tissue plasminogen activator procedure will also be mentioned. Because a controlled trial with a sufficient number of patients and follow-up has not been carried out for any of these techniques, the results of published case series will be used for this review. PMID- 11917795 TI - [Sensory problems after macular translocation]. AB - Following macular translocation, spontaneous compensation of subjectively felt tilt can occur by central adaptation to the rotation of the retinal meridians. However, in cases of large angle rotations or under binocular conditions, efforts to counter-rotate the globe have to be made in order to achieve comfortable vision. For restoring binocular vision postoperatively, the fusionable residual angle of 7-8 degrees should not be exceeded. PMID- 11917796 TI - [Surgical eye muscle counter-rotation after macular translocation]. AB - Surgical counter-rotation after macular translocation is indicated when a non fusionable cyclodeviation exists by lack of suppression. Different surgical options exist to counter-rotate the globe after macular translocation. Primary angles of retinal rotation up to 40 degrees can be compensated for. PMID- 11917798 TI - [Results and complications after deep sclerectomy]. AB - BACKGROUND: As nonperforating glaucoma surgery, deep sclerectomy seems to offer the advantage of fewer complications than the classic trabeculectomy during the first weeks after surgery. PATIENTS AND METHODS: In this prospective study, 74 eyes of 56 patients received deep sclerectomy. The mean follow-up time was 9.5 +/ 5.8 months. Twelve eyes were treated intraoperatively with additional mitomycin C and 11 eyes had combined cataract procedures. The deep sclerectomies were performed without using material of high viscosity or a collagen implant. RESULTS: The mean preoperative pressure of 24.8 +/- 9 mmHg could be lowered to 16.1 +/- 5.9 mmHg (P < 0.0001). The number of glaucoma medications was reduced from 2.2 +/- 1.1 to 0.6 +/- 1.0 substances. Thirty-eight percent of the eyes needed glaucoma medication again. Complications included chorioidal detachment (n = 9), temporary hyphema (n = 6), and delayed pressure reduction (n = 2). CONCLUSIONS: Deep sclerectomy as nonpenetrating glaucoma surgery lowers the intraocular pressure as well as standard trabeculectomy. Its complication rate is very low during the early postoperative weeks. The number of patients who still need glaucoma medication seems to be higher than after trabeculectomy. PMID- 11917797 TI - [Macular translocation with 360 degree retinotomy in the treatment if exudative macular degeneration. Functional and angiographic results]. AB - BACKGROUND: During surgical extraction of choroidal neovascular membranes (CNV) in age-related macular degeneration (AMD), the defective foveal retinal pigment epithelium (RPE) is removed. Subsequent translocation of the foveal neural retina to adjacent healthy RPE should result in stabilization and possibly improvement of visual acuity. METHODS: A prospective case series was carried out using controlled surgery and examination protocols with examinations made at fixed intervals. The surgical procedures combine counterrotation of the globe, phacoemulsification and implantation of a posterior chamber lens, complete vitrectomy, induction of a total retinal detachment, 360 degrees anterior retinotomy, removal of the subfoveal neovascular complex, foveal translocation outside the RPE defect, reattachment of the retina using F6H8, peripheral laser retinopexy and temporary silicone oil tamponade. PATIENTS: Macular translocation surgery was performed on 100 patients between December 1997 and December 1999. All patients had experienced recent visual loss due to exudative AMD and of these, 26 patients had major macular subretinal hemorrhage, 39 patients had occult and 25 patients classic subfoveal choroidal neovascularization. The preoperative findings in the remaining patients included tears in the pigment epithelium (n = 4), polypoidal choroidal vasculopathy (n = 1), recurrent subfoveal CNV following laser therapy (n = 2) and deep retinal vascular anomalous complexes (n = 3). RESULTS: A total of 97 patients completed the 12-month examination. Visual acuity increased by 15 or more ETDRS chart letters in 24 patients, remained stable in 42 patients and deteriorated by 15 or more EDTRS chart letters in 34 patients 12 months postoperatively. The silicone oil tamponade was removed in 97 patients, in 10 patients, silicone oil had to be reinjected because of severe complications. A secondary procedure was necessary in 25 patients, primary PVR was observed in 9 eyes, secondary PVR developed in 10 eyes, a macular pucker in 5 eyes and a macular hole in 1 patient. Other postoperative complications included persistent hypotonia, macular edema, IOL dislocation, keratopathy and recurrent CNV (n = 3). CONCLUSIONS: Macular translocation is a technically demanding operation, which requires a considerable learning curve. Although the procedure has a high rate of surgical and postoperative complications, the functional and anatomical results appear to be promising for selected patients with subfoveal CNV secondary to AMD. PMID- 11917799 TI - [Diffuse lamellar keratitis after laser in situ keratomileusis. Clinical and confocal microscopy findings]. AB - BACKGROUND: Diffuse lamellar keratitis (DLK) is a sterile inflammation after lamellar corneal surgery. A clinical staging for DLK was recently set up. In this observational case series, the clinical and confocal microscopic findings of stages 1-3 are reported. METHODS: Six eyes of six patients (three eyes with DLK stage 1, two eyes with DLK stage 2, and one eye with DLK stage 3) were examined by slit lamp biomicroscopy and confocal microscopy. RESULTS: In all cases, confocal microscopy showed an infiltration of inflammatory cells into the anterior stroma and the flap interface. The number of cells varied between the eyes with DLK stage 1, and both stage 2 corneas had dense infiltrates. In the eye with stage 3 DLK, an aggregation of decayed cells, most likely granulocytes, was noticed clinically and by confocal microscopy. CONCLUSIONS: The DLK stages represent different clinical intensities of interface inflammation after LASIK. While stages 1 and 2 have a similar confocal microscopic appearance, stage 3 is a result of aggregation of a high amount of inflammatory cells. PMID- 11917800 TI - [Corneal neural paralysis after block excision including the 3 and 9 o'clock meridian]. AB - PURPOSE: Corneal innervation is mainly supported by the long posterior ciliary nerves. Anatomically, the long ciliary nerves run with the long ciliary arteries at 3 and 9 o'clock. The aim of this retrospective study was to find out if block excision of anterior uveal tumors or epithelial ingrowth located at the 3 or 9 o'clock position of the limbal circumference causes corneal neuroparalysis. PATIENTS AND METHODS: Between 1980 and May 1999, a total of 151 block excisions were performed in our department (92 block excisions because of anterior uveal tumors and 59 because of cystic epithelial ingrowth to the anterior chamber). In 27 patients, anterior uveal tumors or cystic epithelial ingrowth were located at the 3 or 9 o'clock position of the limbal circumference (14 patients with cystic epithelial ingrowth and 13 patients with anterior uveal tumors). Mean age of all patients was 54.4 +/- 15.7 years at the time of surgery. Mean diameter of the block excision was 10.7 +/- 4.5 mm for tumor-patients and 9.0 +/- 1.2 mm for patients with anterior chamber cysts. Mean follow-up time was 93.6 +/- 43 months. RESULTS: Only 1 of 27 patients exhibited a moderate neuroparalytic corneal ulcer during the follow-up time. In the remaining 26 patients, no signs of clinically relevant corneal neuroparalysis such as epithelial disorders or neuroparalytic ulcers were found. CONCLUSION: Block excision of tumors or cystic epithelial ingrowth located at the 3 or 9 o'clock position of the limbal circumference did not lead to severe neuroparalytic disorders of the host cornea. This may be an important factor in postoperative management of patients undergoing block excision and corneoscleral grafting. PMID- 11917801 TI - [Immunomodulation after keratoplasty by CTL4-Ig and anti-CD154 antibodies]. AB - BACKGROUND: The immunoreaction after corneal transplantation is caused by the T cell receptor interacting with the major histocompatibility complex (MHC) receptor of the antigen-presenting cell. The signal is amplified by the CD4 receptor and the costimulatory signal interactions of CD28-B7 and CD40-CD154. We investigated the influence of costimulatory signal blocking on corneal transplant survival in mice. METHODS: Seven groups of 6 BALB/c mice received an orthotopic corneal transplant from C3H mice differing in minor and major MHC and were postoperatively treated as follows: (1) 80 micrograms of CTLA4 fusion protein intraperitoneally (i.p.) for 6 days; (2) 50 microliters of PBS i.p. for 6 days; (3) 1 mg of Solu-Decortin H i.p. for 5 days + dexamethasone AT 0.1% for 35 days; (4) therapy (3) + 50 micrograms of CTLA4 fusion protein i.p. for 6 days; (5) CTLA4-Ig as in (1) + 15 micrograms of anti-CD154 subconjunctivally (s.c.) on days 0, 2, 4, 6, and 8; (6) CTLA4-Ig as in (1) + 25 micrograms of anti-CD154 s.c. for 9 days; and (7) 25 microliters of PBS s.c. for 9 days. RESULTS: All animals had an immunoreaction on the following days: (1) day 18 +/- 3.1; (2) day 13.6 +/- 1.6; (3) day 48 +/- 6.6; (4) day 65 +/- 41; (5) day 23.5 +/- 8.5; (6) day 16.2 +/ 3.6; (7) day 13.8 +/- 2.7. CONCLUSION: The significant prolongation of transplant survival achieved by corticosteroids alone (P < 0.001) is again significantly increased by combining them with CTLA4-Ig (P < 0.001). Specific immunotherapy combined with nonspecific steroid therapy may also improve clinical corneal transplantation results. Compared to the two control groups, CTLA4-Ig and anti-CD154 only influenced transplant survival at a low dosage (P < 0.001). PMID- 11917802 TI - [Present state of erbium laser phacoemulsification. Comparison with ultrasound phacoemulsification concerning energy and phaco-time at one clinic]. AB - BACKGROUND: Comparison of phaco time and phaco energy between ultrasound (USP) and erbium laser (ELP) phacoemulsification at the Eye Clinic Neubrandenburg. METHOD: One surgeon performed all operations (n = 503) employing only USP (Harmony, D.O.R.C., Groningen, Netherlands). Phaco time was measured and converted to energy. Another surgeon performed all operations (n = 503) using only ELP. The energy transfer and phaco time were read from the Phacolase (Asclepion-Meditec, Jena, Germany). Nucleus hardness distribution was comparable for both surgeons. RESULTS: Phaco times in USP/ELP were as follows: nucleus hardness 0: 0.7 s/7.5 s, nucleus hardness 1: 9.6 s/57.6 s, nucleus hardness 2: 23.7 s/106.9 s, nucleus hardness 3: 39.3 s/215.1 s, and nucleus hardness 4: 68.9 s/232.0 s. Energy in USP/ELP was as follows: nucleus hardness 0: 7.2 J/0.9 J, nucleus hardness 1: 96.7 J/23.5 J, nucleus hardness 2: 237.3 J/53.1 J, nucleus hardness 3: 392.5 J/149.1 J, and nucleus hardness 4: 688.6 J/176.6 J. CONCLUSION: ELP takes longer than USP, but the energy transferred to the eyes for emulsification was lower with the laser in comparison to ultrasound. PMID- 11917803 TI - [The Munich/San Diego/Iowa City Collaboration (MuSIC). MuSIC Report I: Design , characteristics of the collective and preliminary results]. AB - BACKGROUND: We have previously shown that histologically described microcirculation patterns (MCP) can be visualized with indocyanine green (ICG) angiography. We have designed a prospective study to evaluate the prognostic value of these angiographically imaged MCP in small choroidal melanocytic lesions. In this report we describe the design of the study, characterize the patient collective, and present the first results. PATIENTS AND METHODS: In this prospective nonrandomized observational study, unilateral choroidal melanocytic lesions with 1.5-5.5 mm maximum apical height are observed until growth is determined according to defined criteria. Variables are demographic parameters, subjective symptoms, subretinal fluid, location and dimension of tumor, hemorrhage, color, orange pigment, and MCP determined by ICG angiography: normal, straight, parallel without crosslinking, parallel with crosslinking, arcs without branching, arcs with branching, loop, and network. RESULTS: Seventy patients (22 males, 48 females; age: 33-88 years, median: 64 years) have been included up to now: 19 tumors showed growth so far (time to growth: 51-946 days, median: 127 days). The following parameters were statistically significantly correlated with time to tumor growth: flashes (p = 0.082), orange pigment (p = 0.012), subretinal fluid (p < 0.001), maximum basal tumor diameter (p = 0.001), maximum apical tumor height (p < 0.001), parallel with crosslinking (p < 0.001), arcs with branching (p = 0.006), loop (p < 0.001), and network (p < 0.001). Of these, complex MCP (parallel with crosslinking, arcs with branching, loop and/or network) showed the strongest correlation with time to tumor growth in a Cox regression model. Based on our data, the positive predictive value of imaging complex MCP (for growth within 12 months) is 78% and the negative predictive value is 98%. CONCLUSION: Our patient collective demonstrates comparable prognostic parameters for time to growth as described in the literature. In addition, the ICG angiographic detection of complex MCP is more strongly predictive of the time to growth than other clinically determinable factors. Thus, we recommend this examination for patients with small choroidal melanocytic lesions, if the patient is to be counseled regarding the likely biologic behavior of his tumor. PMID- 11917804 TI - [Complications following eyebrow piercing]. AB - BACKGROUND: Piercing as a new form of "body art" is becoming more frequent in western industrialised countries and has achieved a certain social acceptance. Materials used are mainly surgical steel and, less frequently due to higher costs, titanium, gold, silver and niobium. The current literature reports complications such as infections, swellings, allergies, wound healing problems, dental damage and hepatitis following piercing in oral and facial regions. PATIENT: A 22-year-old nurse underwent piercing of her right eyebrow 4 months previously. While piercing of her tongue and nipples had not caused any problems, she first complained of a local inflammation about 3 months after the piercing procedure, which healed spontaneously. Now she complained again about pain, pressure, redness and swelling of her right cheek and face. A dermatologist had recommended tea compresses. On her first visit she presented with a solid, movable, tender, cherry-size swelling of the lateral third of the right eyebrow, a redness of the eyelid and a large swelling of the cheek. Following a systemic antibiotic therapy she underwent surgery and the inflammatory tissue was removed. Histologically, muscle tissue and a piercing canal were visible as well as histiolymphocytotic infiltrates with epitheloid and solitary giant cells. CONCLUSIONS: With an increasing frequency of piercing, the number of complications in the field of ophthalmology is also likely to increase. Considerable costs may arise for the social health care system depending on the severity of the complications. PMID- 11917805 TI - [Bilateral vitreous opacities]. PMID- 11917806 TI - [Paravenous changes in the retinal pigment epithelium]. PMID- 11917807 TI - [Dyslexia. Bases of reading. Reading-writing disorder. Ocular reading disorder]. PMID- 11917808 TI - The global tuberculosis situation. Progress and problems in the 20th century, prospects for the 21st century. AB - Mycobacterium tuberculosis has been identified in prehistoric remains of humans. Despite references to TB by Hippocrates and Galen, humankind had limited understanding of and few tools to defend itself against TB until the later 19th century. Subsequently, landmark advances in the 20th century provided the means to control and prevent this disease. At the same time, epidemiological developments and fundamental problems related to human behavior, socioeconomic conditions, and political circumstances continue to thwart efforts to diminish the burden of suffering and death caused by TB. This article reviewed some of these issues including the global failure of TB control in the late 20th century, the worldwide emergence of drug-resistant TB, the extensive spread of HIV infection and its impact on TB incidence; and changing health care and political environments. The obstacles to TB control remain and will remain challenges in the coming years. Still, recent developments in immunology, biochemistry, and molecular biology suggest that new knowledge and tools are just around the corner. These will enhance the ability to conquer this microbe by the end of the current century. PMID- 11917809 TI - Tuberculosis in patients with HIV infection. AB - In the past 5 years, there have been significant advances in the understanding of the pathogenesis of TB in people infected with HIV and in the approach to diagnosis, treatment, and prevention in patients with HIV. Nucleic acid amplification tests and restriction fragment length polymorphism can contribute to the clinical management of TB patients. New guidelines are available for the treatment of active and latent TB infection in patients with HIV. PMID- 11917810 TI - The mycobacteriology laboratory and new diagnostic techniques. AB - Use of the most rapid and reliable laboratory tests for mycobacterial detection, identification, and susceptibility testing is important for TB control. In 1993, CDC experts made recommendations regarding optimal methods of mycobacterial testing (i.e., stains for AFB, culture, identification, and susceptibility testing of M. tuberculosis) and turnaround times for reporting results. Various technical advances have enhanced the diagnostic capability of the laboratory and/or improved laboratory efficiency since then. The commercial NAA tests for direct detection of MTBC have the greatest potential to impact patient care. To assist physicians, CDC experts have published recommendations concerning use of the NAA tests for management of patients with suspected TB, with emphasis on the MTD assay, which is approved for both AFB smear-positive and smear-negative specimens. With regard to mycobacterial culture, totally automated, nonradiometric systems are commercially available. For mycobacterial identification, various molecular techniques have been developed, but at present, they are used predominantly in research or large reference laboratories. Molecular tests also have proved useful for better understanding the epidemiology of TB and investigating episodes of suspected laboratory cross-contamination. With regard to mycobacterial susceptibility testing, use of the new automated culture systems for testing MTBC is under evaluation, but only one such system has been approved for this purpose. In addition, laboratory guidelines for susceptibility testing of MTBC and certain NTM have recently been published by the NCCLS. PMID- 11917811 TI - Beneficial impact of genome projects on tuberculosis control. AB - The M. tuberculosis genome project is a landmark achievement in the history of TB research. The DNA sequence has provided valuable insights, along with a few surprises, into the complete genetic complement of M. tuberculosis. This information has been used to gain a better understanding of isoniazid-induced alteration in gene expression. It also has been used to construct a genealogy tree of different BCG strains, besides identifying genes that may be responsible for the human-specificity of M. tuberculosis. The impact of this project is far reaching and in the next few years should yield innovative vaccines and therapeutic agents, besides aiding in the rapid and accurate diagnosis of TB. PMID- 11917812 TI - Host susceptibility factors in mycobacterial infection. Genetics and body morphotype. AB - Through identification and evaluation of mutations and polymorphisms in components of the IFN gamma response pathways, a better understanding of the mechanisms and risk factors influencing the development of mycobacterial disease is gained. This may lead the way for development of therapeutic and preventative strategies. Although conventional science has focused on identifying discrete mutations, greater awareness of the impact of subtle changes, both at the genetic (polymorphisms) and physical levels (body morphotype), may prove critical in the investigative process. There has been extraordinary progress in the understanding of mycobacterial susceptibility factors over the last few years. The recognition of characteristic phenotypes will lead to the identification of new genetic bases for disease. PMID- 11917813 TI - Newly described or emerging human species of nontuberculous mycobacteria. AB - The advent of molecular testing in the laboratory has brought about the recognition of multiple newly characterized mycobacterial species not previously recognizable with most standard techniques. Some of the species are nonpathogenic, but the majority may cause clinical disease. Each is likely to have its own biology, drug susceptibility pattern, and response to drug/surgical therapy. Thus, it is important to try to recognize these new species in the laboratory. A study of the phenotypic and genotypic characteristics of these new species also may help to elucidate the epidemiology and pathogenesis of these organisms. In addition, there are multiple emerging species of nontuberculous mycobacteria including M. ulcerans, M. haemophilum, M. xenopi, and M. malmoense. [table: see text] These species are being recognized increasingly as a cause of human disease and recovered within the laboratory. The clinician must learn about these new pathogens to recognize them clinically and assist the laboratory in their recovery. PMID- 11917814 TI - Nontuberculous mycobacteria in cystic fibrosis. AB - The incidence of NTM pulmonary infections increasingly is recognized in patients with CF. This may reflect the increasing longevity of this population with increased environmental exposure time, a high index of suspicion, and/or some as of yet unidentified predisposing factor(s). The most common species of NTM in CF is MAC, followed by M. abscessus. The authors recommend that adult patients with CF be screened for the presence of NTM pulmonary secretions on a regular basis. Positive cultures are likely to indicate disease if they are multiple or if a patient has clinical evidence of pulmonary disease exacerbation (increased cough, increased purulence of secretions, or systemic manifestations such as fever and weight loss) that is not responding to conventional antibiotic therapy. CF patients who do not respond to treatment for the usual organisms should be re evaluated for the presence of NTM and treated with a macrolide-containing regimen directed against the identified NTM if diagnostic criteria are met. Novel treatments with cytokines and intermittent dosing of antibiotics are currently under investigation. PMID- 11917815 TI - Diagnosing nontuberculous mycobacterial lung disease. A process in evolution. AB - Assessing the impact of the diagnosis if NTM lung disease on the patient and choosing appropriate therapy are separate considerations. Health care professionals have progressed from an era when patients had unrecognized, progressive, and untreated NTM disease to an era when NTM disease is diagnosed frequently but therapy is either unnecessary or possibly worse than the disease. Perhaps the 1990 ATS statement was correct. The problem is not diagnosing patients with NTM lung disease, the problem is deciding what to do with them after they are diagnosed. PMID- 11917816 TI - Tuberculosis control in a border state. Treatment of the foreign-born. AB - As the epidemiology of TB in the United States changes, with more foreign-born and fewer native-born residents developing the disease, treatment can be expected to become more complicated and expensive. PMID- 11917817 TI - Multidrug-resistant tuberculosis. AB - Multidrug-resistant TB is a growing public health problem. Although control of the multidrug-resistant TB epidemic has been achieved in New York City, strains of multidrug-resistant TB are found in nearly every state. Much of the world faces a growing problem with no immediate solution. The treatment habits and policies that have led to this problem persist. New drug development has been almost nonexistent. The current tremendous global interest offers hope, as does the Global Alliance for Tuberculosis Drug Development supported by the Rockefeller Foundation and the Bill and Melinda Gates Foundation. Their mission is to accelerate the discovery and development of new antituberculosis drugs and put them on the market within 10 years, at prices affordable in less-developed countries. As Dr. Reichman notes, "this action will not address the main underlying cause of almost all drug resistance, non-adherence of patients and doctors to recommended regimens." He calls for an equal commitment to improving the capacity of health care workers to use new and older agents correctly, so that they may continue to be effective in the future. Each year brings new, at risk immigrants to the United States from all regions of the globe. They bring all the TB problems of their countries of origin with them. Foreign-born people will have a significant impact on TB control efforts in the next decade and beyond. TB elimination programs will need to incorporate systems that can adequately address TB within the United States and support national TB programs in developing countries to help them develop the capacity to successfully manage their own TB problems. A review of the drug resistance patterns isolated from foreign-born people should be adequate to convince even skeptics of the need to support global TB programs. PMID- 11917818 TI - The perils of prescribing. PMID- 11917819 TI - Redbook and PSA testing. PMID- 11917820 TI - The redbook and prostate cancer. PMID- 11917821 TI - Problems of polypharmacy. AB - BACKGROUND: Polypharmacy is common in the management of many conditions. The aim of good prescribing should be to avoid unnecessary polypharmacy. OBJECTIVE: To describe how the World Health Organisation Guide to good prescribing provides a structure for improving prescribing and avoiding polypharmacy. DISCUSSION: Therapeutic goals should be individualized in order to use only those medicines that achieve desirable outcomes for the individual patient. Nonpharmacological approaches should be reviewed as these can result in better patient outcomes and can reduce the dose and the number of medications used. Patients should be monitored for not only the effects but also the adverse effects of prescribed medications. If an adverse reaction is found, the therapeutic approach should be reassessed and alternative approaches or medications triald rather than adding another medication to ameliorate the adverse effects of the first. PMID- 11917822 TI - Chemotherapy and radiotherapy. When to call it quits. AB - BACKGROUND: Patients diagnosed with cancer are often treated with chemotherapy and radiotherapy with curative intent. The transition from curative to palliative intent involves re-evaluation of treatment, and has to take into account the attitudes, beliefs and life aims of the patient. OBJECTIVE: To discuss the difficulties in determining when to cease chemotherapy and radiotherapy in patients with advanced cancer. DISCUSSION: The concept of treatment evaluation using a 'burden versus benefit' paradigm is discussed. Treatment aims must be in concordance with those of the patient, which are often couched in functional terms or linked to future significant life events. Chemotherapy and radiotherapy can offer patients in the palliative phase of cancer illness, benefits in terms of relief of symptoms and meaningful prolongation of life, and should be considered in appropriate circumstances. PMID- 11917823 TI - Antiretroviral medications and HIV. AB - BACKGROUND: Since 1996 the use of combination antiretroviral treatment including protease inhibitors has dramatically improved the prognosis for many people with HIV. However, antiretroviral medications are associated with significant side effects. OBJECTIVE: To outline some of the important side effects of antiretroviral medications and strategies for managing complications. DISCUSSION: Awareness of potential problems, careful prescribing behaviour and close monitoring can reduce the impact of many side effects of antiretroviral medication. Identification and management of side effects may also assist in improving adherence to treatment. PMID- 11917824 TI - A child with earache. Are antibiotics the best treatment? AB - BACKGROUND: Conventional management of acute otitis media, deduced from pathophysiology, embodies scant regard to analgesia, but concentrates on the microbiological cure by the use of antibiotics. OBJECTIVE: A commonly encountered case of uncomplicated acute otitis media in a child of three is presented. An evidence based approach to management is offered. RESULTS: The search for empirical evidence of patient relevant benefits and harms challenges the conventional approach. A Cochrane review of the literature on the use of antibiotics for acute otitis media shows that the benefits of antibiotic use (reduced pain in those children who go on to suffer pain beyond 24 hours) are offset by adverse events from the antibiotics themselves (gastrointestinal symptoms and rashes). There is insufficient information to be sure about rare complications of not using antibiotics as well using them, but it seems that in either case, catastrophe is very rare. DISCUSSION: Thinking in terms of a balance of harms and benefits would result in a decreased proportion of children prescribed antibiotics for acute otitis media. PMID- 11917825 TI - Influenza. Can we vanquish this foe? PMID- 11917826 TI - Flu facts. What is influenza? AB - Influenza, commonly called 'the flu', is an illness caused by the influenza virus. The virus is passed form person to person by sneezing or coughing. Typical symptoms of influenza include fever, cough, sore throat, fatigue, muscle aches, headaches, runny nose and watery eyes. PMID- 11917827 TI - An easy way to inject the knee joint. PMID- 11917828 TI - Taping. General principles. AB - This is the first in a series of three articles outlining taping techniques on a regional basis. Where taping has a role in the management of an injury, the articles also describe appropriate site specific techniques. Further, taping of the more common conditions encountered in general practice are described. PMID- 11917829 TI - Management of bedwetting. AB - BACKGROUND: Nocturnal enuresis (bedwetting) is a common inherited medical condition affecting both children and adults. It is not due to laziness. It can be distressing, embarrassing, humiliating and has a negative impact on self esteem and mood especially in children and young people over the age of 10 years. OBJECTIVE: To assist busy general practitioners develop a better understanding of bedwetting and the importance of its diagnosis and treatment. DISCUSSION: Spontaneous remission rates over the age of 10 years are low at 5% per year and approaches often tried at home such as waking to void, reward charts or fluid restriction are not effective. However, treatment with alarm systems or desmopressin for those who fail to respond to alarms is successful in over 90% of cases. PMID- 11917830 TI - Transcendental meditation, hypertension and heart disease. AB - BACKGROUND: Accumulating evidence that stress contributes to the pathogenesis and expression of coronary heart disease has led to the increasing use of stress reduction techniques in its prevention and treatment. The most widely used and tested technique is transcendental meditation. OBJECT: To describe transcendental meditation and review research on its use in the treatment and prevention of coronary heart disease. DISCUSSION: Transcendental meditation shows promise as a preventive and treatment method for coronary heart disease. Transcendental meditation is associated with decreased hypertension and atherosclerosis, improvements in patients with heart disease, decreased hospitalisation rates and improvements in other risk factors including decreased smoking and cholesterol. These findings cannot be generalised to all meditation and stress reduction techniques as each technique differs in its effects. Further research is needed to delineate the mechanisms involved and to verify preliminary findings concerning atherosclerosis and heart disease and the findings of short term hypertension studies. PMID- 11917831 TI - The difficult patient. AB - A large percentage of any general practitioner's time is taken up by a small number of patients, the 'difficult' patient. These patients often have chronic, painful problems, and are unresponsive to traditional medical methods. By understanding the concepts of the patient centred clinical method, and looking beyond the traditional biological model of disease, an approach to these patients can be developed that is simple and enduring. PMID- 11917832 TI - Doctor as patient. Part 2: The realities of life as a patient. PMID- 11917833 TI - Medical fees. Medibank and Medicare. AB - Because paying the doctor was often a financial burden, many ways were devised to help pay the doctor's bill. This article looks at how private medical insurance began to merge into a government funded medical practice. Recent experiences suggest that payment for general practitioners is about to undergo further change. PMID- 11917834 TI - General practice research. For universities only, or a normal part of everyday care? PMID- 11917835 TI - Primary care research networks for Australia? PMID- 11917836 TI - Communication across the divide. A trial of structured communication between general practice and emergency departments. AB - AIM: To evaluate the impact of structured form letters for general practitioner to emergency department (ED) communication. METHOD: STUDY POPULATION: one hundred and fifty-five GPs with practices in the Liverpool local government area in metropolitan Sydney and patients referred by them to ED at Liverpool over five months from June to October 1998. DESIGN: randomised control trial of GPs as unit of randomisation; intervention GPs were encouraged to follow a structured proforma for their written communication with the ED. Control GPs were left to usual referral procedures. The ED was encouraged to fax a brief report back to GPs using the form. Impact measures: the quality of the referral letters was evaluated using a checklist that included: reason for referral; examination finding; medical history; investigations; psychosocial history; allergies; drugs given in the surgery and present medication. Surveys were sent every month to GPs to assess communication from the ED and adverse events observed by GPs. RESULTS: Most letters from GPs to the ED contained information on reasons for referral, medical history and examination findings. Reasons for referral were present in 95% of the intervention group GPs' letters compared with 99% of those of the control group. Investigations were included with 27% and present medications in 37%. Letters from GPs in the intervention group were more likely to contain a psychosocial history than those in the control group (13% compared with 1%). Most GPs reported receiving a letter from the ED although this was rarely by fax; most were brought to them by the patient. Phone calls were received by about one in five GPs each month. Most GPs found both of these to be useful. There were no differences between communication received by GPs in the intervention and control groups. CONCLUSION: This study demonstrates that improvements to communication between GPs and EDs are difficult and may require a systemic change within general practice and the hospital. Electronic systems may allow the sort of reciprocal communication required to establish and sustain improvement. PMID- 11917837 TI - Research capacity building in general practice. The new Australian scene. AB - BACKGROUND: Primary care research needs strengthening. The Commonwealth Government Department of Health and Aged Care has recently funded the university departments of general practice and rural health to build research capacity in primary care. OBJECTIVE: To explore issues surrounding building primary care research capacity, as well as looking at barriers to research capacity building and ways of overcoming them. DISCUSSION: New funding provides many opportunities for increasing research capacity in primary health care areas. Different institutions will select those methods that are best suited to their skills and the requirements of their area. PMID- 11917838 TI - Research capacity in general practice. Opinions from the field. AB - BACKGROUND: The Primary Health Care Research Evaluation and Development (PHCRED) Strategy includes funding for universities to build research capacity. As part of its implementation, the University of NSW consulted widely with Divisions of General Practice, GPs and representatives from Community Health Services to determine current research capacity. OBJECTIVE: We summarise the results of this consultation, and outline implications for the PHCRED Strategy. DISCUSSION: Divisions reported low research capacity, and welcomed the opportunity to develop it further. GPs with an interest in research were particularly focused on clinical areas, and saw their role in providing patients and collecting data, rather than initiating and developing ideas. Workers in Community Health Services were keen to be involved. There was considerable support and enthusiasm for the concept of capacity building and the activities proposed, despite perceived low levels of research skills. The overall aim of the PHC-RED Strategy may need to be clarified to successfully build on this enthusiasm. PMID- 11917839 TI - [Investigation on root-knot diseases of medicinal plants in Yunnan]. AB - During 1994-1997, the damage levels about root-knot diseases on 52 medicinal plants were investigated and analyzed. 185 samples were collected from more than 20 counties or cities in Yunnan province. The results showed that 76 specimens were infected by root-knot nematode, which are Meloidogyne incognita (Kofoid and White 1919, Chitwood 1949), M. javanice (Trenb 1855, Chitwood 1949), M. arenaria (Neal 1889, Chitwood 1949) and M. halpa (Chitwood 1949). They infected 21 medicinal plants. The paper describes the damage levels and the distribution about these root-knot diseases of medicinal plants. PMID- 11917840 TI - [The species and the distribution of officinal nettles in Sichuan Province]. AB - OBJECTIVE: To study the resource of officinal nettles in Sichuan. METHODS: Investigating at the wild spots and consulting the relating specimens. RESULTS: It has been found that there are 11 spieces and subspieces of genus Urtica in Sichuan Province, of which 8 nettles are commonly used for treatment of rheumatism. CONCLUSION: This paper can provide scientific reference for further study of officinal nettles in Sichuan. PMID- 11917842 TI - [The identification of flos magnoliae by TLC]. AB - OBJECTIVE: To establish a TLC identification method for Flos Magnoliae. METHOD: Selecting optimum TLC conditions. RESULTS: A TLC identification method of Flos Magnoliae has been established. CONCLUSION: The method is simple, accurate and reliable. PMID- 11917841 TI - [Investigation on method speliality in identifying Fructus Crataegi with vitexin rhamnoside]. AB - The methanol extract of Fructus Crataegi was purified on macro-reticular resins(model DM130)-polyamide column. Vitexin rhamnoside(I) was identified from Fructus Crataegi by TLC on polyamide layer sheets. The false and the confused species of Fructus Crataegi and some traditional Chinese herbal medicines which contain vitexin were studied by the same method. The results indicated that compound I was special constitute of Fructus Crataegi and the identification method of Fructus Crataegi with I was special. PMID- 11917843 TI - [Studies on chemical constituents from the flower of Citrus aurantium]. AB - The chemical constituents from the flower of Citrus aurantium were studied. 11 compounds were isolated and identified including neohesperidin(I), synephrin(II), 5,8-epidioxyergosta-6,22-dien-3 beta-ol(III), adenosine(IV), asparagine(V), tyrosine(VI), valine(VII), isoleucine(VIII), alanine(IX),beta-sitosterol(X) and beta-daucosterol(XI). PMID- 11917844 TI - [Effects of pyracantha fortuneana extract on blood coagulation]. AB - OBJECTIVE: To study the effect of the Pyracantha fortuneana extract and its defferent polar fractions on blood coagulation. METHODS AND RESULTS: The Pyracantha fortuneana extract could noticeably shorten the coagulation time of glass slide in mice (P < 0.05) after oral administration, and also shorten the recalcification time and prothrombin time in vitro (P < 0.01, P < 0.05), while in three polar fractions of the extract, the effects of the chloroform phase that could shorten the coagulation time of glass slide in mice(P < 0.01) after oral administration, and also shorten the recalcification time and prothrombin time in vitro (P < 0.01) are more noticeable than that of the acetic ether phase (P < 0.01, P < 0.05). The petroleum ether phase has no above-mentioned effect (P > 0.05). CONCLUSION: The Pyracantha fortuneana extract can reduce the blood coagulation. Both the chloroform phase and the acetic ether phase of the extract have activity of blood coagulation, but the petroleum ether phase does not. PMID- 11917845 TI - [Study on apoptosis of human gastric adenocarcinoma cell SGC-7901 induced by aining in vitro and the cell cycle]. AB - OBJECTIVE: To study the inducing apoptosis effect of Aining on human gastric adenocarcinoma cell in vitro. METHODS: Agarose gel electrophoresis (AGE) and flow cytometry (FMC) techniques were adopted to study the inducing apoptosis effect of Ainging on the human gastric adenocarcinoma cell. RESULTS: The human gastric adenocarcinoma cell's DNA was digested into fragments which gave rise to a characteristic "DNA Ladder" on agarose gel electrophoresis, while the percentage of apoptotic cells was 43.5%. But the blank group had neither DNA ladder, nor apoptotic peak. Cell cycle analysis showed that the proliferation index was 33.7% after the human gastric adenocarcinoma cells treated with Aining, and there was no cell phase specialty. CONCLUSION: Aining can induce human gastric adenocarcinoma cell apoptosis in vitro. PMID- 11917846 TI - [Study on immunonodulating activities of enzymolyzed loach protein]. AB - The immunomodulating activities of enzymolyzed loach protein in mice were investigated by regular methods for immunology evaluation. The doses of enzymolyzed loach protein were 0.1, 0.2, 0.4 g.kg-1 bw, which were 5, 10, 20 times respectively as large as the recommended daily intake(0.02 g.kg-1 bw). After being successively fed with enzymolyzed loach protein at given doses for 30 days by ig respectively, the mice were detected by immunomodulation tests. The results indicated that enzymolyzed loach protein had significant modulating effects on all of test indexes and suggested it had apparent immunomodulating activities in mice. Based on the results described above, we speculate that the enzymolyzed loach protein may have immunomodulating functions and can enhance the immunity of body. PMID- 11917847 TI - [The antioxidative effect of procyanidins from pine bark in vitro]. AB - OBJECTIVE: To assess the antioxidative effect of procyanidins from pine bark on free radical damage. METHODS: Hemolysis, Malonaldehyde(MDA) level of mice liver homogenates, the conformation changes of irradiated plasmid PUC18 were used as indexes. RESULTS: Procyanidins could reduce the hemolysis degree of human RBC induced by H2O2 significantly(P < 0.01), reduce the MDA level of mice liver homogenate initiated by VitC/Fe2+ remarkably(P < 0.01), and reduce the degree of single-strain break of plamid PUC18 induced by 60Co gamma significantly(P < 0.01). CONCLUSION: Procyanidins had good antioxidative function. It can prevent RBC membrane, plasmid DNA suffering from oxygen free radical damage in vitro. PMID- 11917848 TI - [Pharmacodynamics study of suanzaoren decoction extracted by different technological process]. AB - OBJECTIVE: To select the optimal technological process for Suanzaoren decoction. METHOD: The effects of extracts from different technological processes on spontaneous motion and sleep time in mice induced by pentobarbital sodium (subthreshold dosage) were studied. RESULTS: Both the decoction and 95% alcohol extract of Suanzaoren possess sedative and hypnotic effects. CONCLUSION: Suanzaoren aqueous decoction was selected as the optimal technological process. PMID- 11917849 TI - [Protective effect of huyin decoction on acute hepatic injury induced by CCl4 in mice]. AB - OBJECTIVE: To observe the protective effect of Huyin decoction on acute hepatic injury induced by CCl4 in mice. METHODS: The model of acute hepatic injury induced by CCl4 was used. The levels of ALT and AST in serum were detected. And histopathology of liver was observed. RESULTS: Huyin decoction could decrease ALT and AST in serum and hepatic histopathological injury of the acute hepatic injury mice. CONCLUSION: Huyin decoction possessed protective effect on acute hepatic injury in mice. PMID- 11917850 TI - [Effects of cervical vertebra rehabilitation pill on antiinflammation and analgesia]. AB - OBJECTIVE: To study the effects of cervical vertebra rehabilitation pill on anti inflammation and analgesia. METHODS: Hot board and body distortion induced by acertic acid methods were applied in mice for analgesia experiment. Granuloma induced by cotton ball implantation tumefaction in feet induced by carrageenin in rats and auricle tumefaction induced by croton oil in mice were applied for anti flammation experiment. RESULTS: Cervical vertebra rehabilitation pill could significantly prolong the latent period of pain reaction induced by hot board in mice and markedly decrease the frequency of body distortion induced by acertic acid. Cervical vertebra rehabilitation pill also could significantly inhibit the tumefaction in feet induced by carrageenin, reduce the degree of tumefaction in auricle induced by croton oil in mice and inhibit the formation of granuloma induced by cotton ball implantation in subcutaneous part. CONCLUSION: Cervical vertebra rehabilitation pill has significant effects on anti-inflammation and analgesia. PMID- 11917851 TI - [Selection of optimal ultrasonic extraction process of Elaeagnus angustifolia L. by uniform design]. AB - The influences of ultrasonic frequency, ultrasonic time and solvent content on the extraction rate of fat oil in Elaeagnus angustifolia L. were studied. The optimum extraction condition was obtained. The results showed that the ultrasonic extraction method of fat oil in Elaeagnus angustifolia L. could save time, improve extraction rate and need not be hot compared with traditional extraction method. PMID- 11917852 TI - [Determination of isorhamnetine and quercetin in flavone hippophaes tablets by HPLC method]. AB - OBJECTIVE: A method was established for the determination of isorhamnetine and quercetin in flavone hippophaes tablets by HPLC. METHODS: The HPLC system consisted of Diamonsil C18 column (250 mm x 4.6 mm, 5 microns), mobile phase of methanol-0.02 mol/L phosphoric acid (54:46), column temperature at 40 degrees C, with detection at 368 nm. RESULTS: The linear range of isorhamnetine and quercetin were 6.5-32 micrograms/ml (r = 0.9996) and 2.5-12.4 micrograms/ml (r = 0.9991). The average recoveries were 99.4% and 99.8%, respectively. The within day precision RSD were 1.22% and 1.32% (n = 6), and the day-to-day precision RSD were 1.07% and 1.35% (n = 5), respectively. CONCLUSION: This method is simple and accurate. It is suitable for the preparation quality control. PMID- 11917853 TI - [Technical causes of false and inferior species appearing in Chinese medicinal materials markets]. AB - The result indicated that technical causes of false and inferior appearing in the Chinese medicinal materials markets consisted of two respects essentially. First, the men who violated the law and committed a crime used certain weak link existing in quantity analysis of country's medicinal standard. Second, it is difficult that basic units managing and using drug implement effective technical supervision. PMID- 11917854 TI - [Pulmonary chondromas and the Carney triad]. PMID- 11917855 TI - [Understanding of infection in the 21st century]. PMID- 11917856 TI - [Infection of a global scale]. PMID- 11917857 TI - [Immunological mechanism in man against infection]. PMID- 11917858 TI - [Bacterial acquisition of multiple drug resistance and changes in their pathogenicity]. PMID- 11917859 TI - [Infection in immunocompromised host and its management--with special reference to respiratory tract infections]. PMID- 11917860 TI - [Objective in the strategy for prophylactic vaccination]. PMID- 11917861 TI - [Direction in the development of antibacterial drugs in the age of genomic analysis]. PMID- 11917862 TI - [The roles of the infection control doctor (ICD) and infection control nurse (ICN)]. PMID- 11917863 TI - [Provirus and diseases]. PMID- 11917864 TI - [Chlamydia pneumoniae infection and arteriosclerosis]. PMID- 11917865 TI - [Autoimmune peripheral nerve diseases following infection]. PMID- 11917866 TI - [Versatility of EB virus infection]. PMID- 11917867 TI - [Progression of HBV and HCV hepatitis into the chronic stage and cancer]. PMID- 11917868 TI - [Chronic fatigue syndrome (CFS)]. PMID- 11917869 TI - [Airway biofilm disease and autoimmunity]. PMID- 11917870 TI - [Correlation among infection, immunity, and hypersensitivity: discussion]. PMID- 11917871 TI - [Treatment of Crohn disease--with special reference to anti-TNF antibody therapy]. PMID- 11917872 TI - [Organ resuscitation--resuscitation medicine]. PMID- 11917873 TI - [Patient care and ethics. VI. Expensive treatments and the very old patient]. AB - On the basis of a case report in which a 92-year-old patient requests treatment with dental implants, the question is raised whether there is a moral justification to deny the wishes of the patient. PMID- 11917874 TI - [Determining factors for retention of complete dentures]. AB - Patients with full dentures often experience some problems with their dentures, one of them is retention. The retention of a full denture is a result of many factors. Some factors are of a physical nature, others of a physiological nature. The dentist has to take these factors into account while modelling the borders, flanks and basis of the prosthesis. In obtaining the best shape of these borders, flanks and basis a special emphasis is put on the border and flank modelling and the individual impression. PMID- 11917875 TI - [Complete dentures in the Dutch population 1981-1992]. AB - In the period 1981-1992 the percentage of people with a full set of dentures dropped from 31.6 to 22.5 in the Netherlands (persons > or = 16 years). This is particularly due to the relatively strong decrease in the number of young people with full dentures. Regional differences are remarkable: in the northern provinces the percentage is relatively high; in the province of Utrecht it is relatively low. Recently regional differences appear to be (very) small among the young age groups. Many people with a full set of dentures use the same dentures for a long period: about 20% of them have had their current prostheses for 16-25 years, another 20% for over 25 years. Taking into account the total adult population, it appears that the socio-economic status (SES) negatively correlates with the probability of having a full set of dentures. This holds for all three SES indicators in the study: education, income and type of insurance. But among the young age groups this correlation is only valid for education. PMID- 11917876 TI - [Education on the subject of ethnic minorities]. AB - Taking into account the large number of patients belonging to ethnic minorities (in the four big cities they represent about 25% of the population) and the problems healthworkers often experience in treating these patients, it is desirable that future dentists and dental hygienists are prepared at their school to communicate with members of minority groups. Attention should be paid to the attitude of the student towards members of ethnic minorities as well as to the enlargement of knowledge (of demography, different patterns of diseases, social cultural background, identity problems, attitude towards healthworkers and more). The attention paid in the curriculum of dental schools (dental students and dental hygienists) however, is (nearly) nil, with the exception of the Amsterdam dental hygienist school. PMID- 11917877 TI - [Dental caries in children from ethnic groups in the Netherlands. A literature review]. AB - This article reviews the literature on child dental caries in various ethnic groups in the Netherlands. Material from fifteen studies showed generally the dental health of five- to eleven-year-old children of parents born in Turkey or Morocco to be far worse than that of children of Dutch or Surinamese parents. The ethnic groups distinguished, however, turned out not to be homogeneous with regard to dental health. Within each ethnic group the socio-economic status as well as the parental level of fluency in the Dutch language were important additional risk indicators for caries. Regular toothbrushing seemed to be the most important factor to diminish the caries risk. PMID- 11917878 TI - [Prevention of osteoporosis and pathological calcifications]. AB - Based on physiological data about the influence of magnesium compounds on the degree of saturation of blood plasma with calcium phosphates, we have hypothesized in 1988 that magnesium deficiency is the real cause of osteoporosis and pathological calcifications. The incidence of these diseases is among the elderly as high as 40%. These diseases are the reason that there is such a discrepancy between the morbidity and mortality curve for the elderly as a function of age. Meanwhile it has been proven that a. in all osteoporotic patients the intracellular magnesium content of the erythrocytes is lower than in controls, b. this content can be restored to normal by oral magnesium supplementation without increasing the serum magnesium level, c. by this supplementation the bone density in perimenopausal women increases and d. this supplementation keeps the blood plasma and probably also the other extracellular fluids undersaturated with octocalcium phosphate so that it is impossible for pathological calcifications to occur or to progress. The possible benefits for dentistry are the following: a. perhaps mandibular resorption is retarded or inhibited by oral magnesium supplementation and b. this treatment may also help to inhibit or diminish dental calculus formation in heavy calculus formers. Since both aspects are accessible to experimental clinical investigation, we will soon have the answer to these questions. PMID- 11917879 TI - [Dento-alveolar cleft and the lateral incisor]. AB - In the period of 1980 to 1990, 55 cleft, lip and palate patients, ranging in age from nine to twelve years, were treated. With 52% of them the lateral incisor on the cleft side was agenetic, in 24% normally developed and in 24% malformed. The malformed and the fissural teeth were removed during bone grafting of the dento alveolar cleft. In almost all cases a well-formed tooth arch could be obtained by orthodontic treatment. PMID- 11917880 TI - [Ethnicity and the 'TJZ' decision]. AB - The aim of this study was to assess differences in oral health and dental behaviour in a cohort of five-year-old children born in 1982 and a cohort of eleven-year-old children born in 1976, in different ethnic and socio-economic groups, in the period from 1987 to 1990. To this end a secondary analysis was performed on data collected in a study monitoring the oral health of sick fund insured youths in The Netherlands. Large differences in dental health and dental health behaviour were found between ethnic and socio-economic strata. Thus, in the 1982 cohort, the caries prevalence and incidence were highest in the stratum of children of Turkish or Moroccan mothers with a low or unknown education. In the 1976 cohort, the caries prevalence and incidence were highest in children of Dutch parents with a low or unknown education. It is concluded that both stratification variables were caries risk indicators. PMID- 11917881 TI - [Patient care and ethics. VII. Ethics education in the university dental curriculum in the Netherlands]. AB - Dentistry, which has developed independently alongside medicine for more than a hundred years now in the Netherlands, is rooted in a type of natural scientific and technological thinking that gained wide acceptance in the middle of the last century. This type of thinking has had far-reaching consequences for the whole field of medicine, including the independent development of dentistry. The social science side of dental training and the normative aspects of clinical practice have long been seriously neglected as a result of the dominance of the natural sciences. This article surveys development in medicine and dentistry since the middle of the last century. It then examines recent developments in medicine and dentistry and concludes by describing the current status of education in ethics in the dentistry curriculum. PMID- 11917882 TI - [Adhesion of composite to dentin. Mechanical and clinical results]. AB - Although an efficient bond of resin composites to enamel can be realized since quite a long time, reliable dentine bonding is nowadays still a clinical problem. After the failure of the dentine-etch technique, followed by the misfortunes of the chemical dentine adhesion technique, modern dentine adhesive systems are believed to function by a micromechanical attachment mechanism. Based on a morphological study of the resin-dentine interface, a broad selection of dentine adhesive systems was classified following their adhesion-strategy. In a second part, eight dentine adhesive systems were clinically tested in terms of retention. PMID- 11917883 TI - [Statistics for dentists (VIII). Student t-test]. AB - In order to test whether the mean scores of either two paired or independent samples deviate from one another, the Student t-test could be applied. In the paired samples case, the mean score of the differences between the matched scores is analyzed. In case of two independent samples, the difference between the two means is analyzed. In both cases the t-value is the difference divided by the error in difference, which, in combination with the degrees of freedom, results in the p-value. PMID- 11917884 TI - [Proceedings of 31st annual meeting of the Japanese Society of Neuropsychopharmacology. Hiroshima, Japan. October 4-5, 2001. Abstracts]. PMID- 11917885 TI - Consumption of food products by the elderly living in different environments of the Warsaw region, Poland. AB - The aim of the study was to compare qualitatively the intake of food groups by the elderly living in urban and rural areas, with regard to the place of living and sex. Fifty-sex subjects in Warsaw and 142 in villages (born in 1919-24) participated in the study. A 3-day record method was used to collect nutritional data. The subjects living in Warsaw consumed more often milk, meat, fish, fruit and their products, as well as fats and oils. Less than 50% of the elderly living in either rural or urban areas ate eggs and fish. Consumption of fruit and its products was also low in the villages (47%). Meat and meat products were consumed more often by elderly men, whereas milk, fruit and their products were eaten more often by women (a rural area only). Consumption of specific food items also differed between the two analyzed living areas; milk, yoghurt and cheese were eaten more often by the residents of Warsaw, however, the intake of cream was significantly higher in the villages. The intake of red meat and meat products was similar, but poultry was included into the diet of 24% of the elderly in the rural area, significantly less than in Warsaw (34%). It also appeared that butter and mixed fats were more common among the elderly in Warsaw,whereas lard, pork fat and margarine were more popular in the villages. Food choice in the rural area was of poorer quality. PMID- 11917886 TI - Does the source of support matter for different health outcomes? Findings from the Normative Aging Study. AB - OBJECTIVES: This study investigated the differential relationships between different types and sources of social support and physical and mental health. METHODS: Using data from the Normative Aging Study, 1,386 older men (median age = 62.7 years) were categorized into four groups separately for frequency of interaction with networks and perceived support. RESULTS: More than half the sample reported high levels of support from both sources. One-way ANOVAs revealed that those with high perceived support from both sources reported better physical health and fewer depressive symptoms than those with low support from both sources or high support from family alone. Similarly, those with high perceived support from both sources had lower levels of depressive symptoms than those with low support from both sources, but frequency of contact was unrelated to physical health. DISCUSSION: In general, those with high support from both family and friends reported the highest level of well-being. PMID- 11917887 TI - Re: "Demonstration of epidermal transfer from a polymer membrane using genetically marked porcine keratinocytes'. PMID- 11917888 TI - [Regional meeting of the Japanese Society of Otolaryngology. Japan. 2001. Abstracts]. PMID- 11917889 TI - [Runny nose, headache and fever in a football player--an allergy?]. PMID- 11917890 TI - [Odd side effects of an analgesic]. PMID- 11917891 TI - External canal cholesteatoma. PMID- 11917892 TI - Committee reports. Report of the Ad hoc Committee on the chemically dependent dentist. PMID- 11917894 TI - New star system for the NHS and style of handling poor performance--a significant step forward for improving the NHS. PMID- 11917895 TI - Older people brand government nursing plans--a broken promise. PMID- 11917896 TI - Disguising medication. PMID- 11917897 TI - PFI extended into primary care into poorest parts of England--new cash injection to improve GP surgeries. PMID- 11917898 TI - One million older people face brunt of care with little support from health or social services. PMID- 11917899 TI - Hospitals shine in latest cleanliness checks: WRVS invited on board to keep standards high. PMID- 11917900 TI - UKCC sets out options for the future of nursing education. PMID- 11917901 TI - Package to boost GP numbers and improve working lives in primary care. PMID- 11917902 TI - [Diagnosis of pulmonary artery embolism. part 2]. PMID- 11917903 TI - Salvaging the ischaemic penumbra: more than just reperfusion? AB - 1. The ischaemic penumbra is defined as a moderately hypoperfused region that retains structural integrity but has lost function. In animal models of ischaemic stroke, this region is prone to recurrent anoxic depolarization and will become infarcted if reperfusion does not occur. In the macaque model, an ischaemic penumbra has been identified for up to 3 h after ischaemic stroke onset, whereas in selected human patients it may exist for up to 48 h. 2. Although most definitions of the ischaemic penumbra stress a time-brain volume concept, few incorporate the idea that selective and delayed neuronal injury plays an important role. Thus, in addition to necrotic cell death caused by acute injury, it is important to also consider delayed death mediated by caspase-dependent and independent apoptotic pathways. 3. Salvage of penumbral tissue is possible if reperfusion (e.g. after thrombolysis) occurs. However, neurons within this salvaged region may be still at risk of further delayed neuronal injury. 4. In the present review, we aim to revisit the concept of the ischaemic penumbra and explore the role of selective and delayed neuronal injury in enlargement of the volume of infarction, as well as pathogenic mechanisms of white matter ischaemia. Both animal and human models of cerebral ischaemia imaged using magnetic resonance and positron emission tomography techniques will be discussed. PMID- 11917904 TI - Muscle strength and ageing: methodological aspects of isokinetic dynamometry and androgen administration. AB - 1. To evaluate interventions aiming to increase muscle strength in older men, it is necessary to use an objective and reproducible method to measure strength. The most reliable method to evaluate muscle strength is isokinetic peak torque (PT) measured by a dynamometer; however, raw PT varies with differences in body size and an optimal scaling for body physique to control confounding effects of body size is necessary to make the most valid comparisons. 2. The present study was designed to estimate the effects of age (part A) and androgen administration (part B) on muscle strength and to estimate reproducibility and evaluate various scaling methods in order to optimize comparisons of isokinetic PT measurements. 3. A single isokinetic exercise protocol was used to compare the muscle strength of 31 healthy men of two age groups (< 40 and > 60 years; part A) and change in strength due to administration of dihydrotestosterone (DHT; 70 mg/day) or placebo gel for 3 months in 35 healthy older (> or = 60 years) men (part B). 4. Muscle strength was assessed by a total of 16 PT measurements using a Cybex NORM dynamometer (Cybex, Ronkonkoma, NY, USA). Age-related differences in muscle strength were estimated by using PT evaluated as raw data or scaled by normalizing methods, including simple ratio (PT/weight), allometric PT (PT/weight(0.67)) and adjustment of PT by weight, height, body mass index and body surface area (BSA). The goodness-of-fit for various scaling methods was compared using the Akaike Information Criterion (AIC) as an objective measure of model-based entropy reduction. 5. The effects of DHT administration according to different scaling methods were estimated by eta-squared measure of effect size in treatment models. In part A, older men were weaker than younger men in five knee PT, consistently by all eight analysis models but not in shoulder PT. In part B, DHT treatment resulted in an increase one knee PT (dominant knee flexion at 120 degrees/s) with the difference consistent in all seven models. 6. The scaling model using BSA proved superior to other comparison models throughout both parts of the present study according to entropy minimization criteria (AIC) for goodness-of-fit of the model or eta-squares for treatment effect size. 7. We conclude that differences in muscle strength due to age or androgen administration in older men are restricted to a minority of lower limb contractions and that use of BSA scaling for PT values is considered the best scaling method for muscle strength comparisons in either cross-sectional or longitudinal studies. PMID- 11917905 TI - Pharmacokinetics of organic anions in rats with arterial calcinosis. AB - 1. Ageing induces calcium accumulation in the vascular system. The simplest experimental way of producing high degrees of arterial calcium overload is by administration of an overdose of vitamin D(3) to rats. The aim of the present study was to evaluate the pharmacokinetics of organic anions in rats with arterial calcinosis induced by an overdose of vitamin D(3). 2. We used bromosulfophthalein (BSP) and sulfanilamide (SA) as models of organic anions with preferential biliary and renal excretion, respectively. 3. Increases in the clearance and elimination rate constant of BSP were observed in treated rats. The clearance and the elimination rate constant for SA were also increased in rats with arterial calcinosis. 4. Variations in arterial hepatic blood flow, aspartate aminotransferase activity and liver calcium accumulation were not observed in treated rats. In contrast, treated rats had a lower renal blood flow and increased renal calcium levels. 5. In summary, rats with arterial calcinosis showed an increase in total body clearance of both BSP and SA, probably associated with modifications in their metabolism and/or in organ extraction. Alterations to hepatic and renal blood flow do not account for these phenomena. PMID- 11917906 TI - Enterogastric brake in rats with segmental bowel resection: role of capsaicin sensitive nerves. AB - 1. Unabsorbed nutrients in the distal gut inhibit upper gastrointestinal motility. 2. The aim of the present study was to investigate changes in gastric motility following segmental resections and to evaluate the role of capsaicin sensitive afferent neurons that may be responsible for these changes. 3. Wistar albino rats of both sexes (200-250 g) were used. Under aseptic conditions and anaesthesia (100 mg/kg ketamine), a baby-feeding tube was placed distal to the resection in either the ileum or caecum for intraluminal perfusion of saline or 20 lipid. In one group of rats, capsaicin was perfused (0.6) for afferent denervation. One group of rats underwent jejunal and ileal resections with end-to end anastomosis of the remaining segments and were cannulated distal to these anastomosis. Ten days after the surgery, the percentage gastric emptying of a solid meal was calculated. 4. Intra-ileal (18) and intracaecal (34) lipid perfusions delayed gastric emptying compared with groups perfused with saline (54 and 74, respectively; P< 0.001 and P< 0.01). The delay in gastric emptying by ileal perfusion was significantly greater than that following caecal perfusion (P< 0.05). With both resections, gastric emptying was delayed compared with sham groups (P< 0.05-0.01). Local administration of capsaicin abolished the inhibitory effect of lipid on gastric emptying in healthy intact rats and in the jejunal resection group, whereas a partial reversal was seen in the ileal-resection group. 5. In conclusion, the present data demonstrate that in both healthy intact rat groups and in short bowel groups, capsaicin-sensitive extrinsic neurons participate in brake mechanisms of the distal gut. PMID- 11917907 TI - The cellular origin of plasma cytokine expression after acute exercise. PMID- 11917908 TI - [Present status and future of vestibular function tests]. PMID- 11917909 TI - [14th spring meeting of the Japanese Society of Allergology. Chiba, Japan. March 21-23, 2002. Abstracts]. PMID- 11917910 TI - [Effect of seizure site and EEG anomalies on cognitive function in epilepsy in the child]. PMID- 11917912 TI - [Grmek, the "thunderbolt", "thunder-clapp" (as testimony)]. AB - The paper details how the author (who was a young physician) met, ... by a mere chance. Prof. Grmek a "Sorbonne", in November 1978, how he felt the "Lecons du Jeudi" at the Ecole Pratique des Hautes Etudes (Fourth Section : "Historical and Philological Sciences"). Grmek was both Croatian and French. Other testimonies are given coming from the disciple (who had Prof. Grmek as director of thesis at Paris I - Pantheon - Sorbonne, in 1986) and coming also from Prof. Vera Gavrilovic (Serbia) who met Grmek when he as in "Yugoslavia". Grmek's quotings and photographies, devoted to this exceptional searcher in the field of History of Medicine and History of Sciences, are reported and showed. PMID- 11917913 TI - [Grmek as scholarly editor and director]. PMID- 11917914 TI - [Professor Mirko Drazen Grmek and the French Society of the History of Medicine]. PMID- 11917915 TI - [Mirko Grmek, medical friend and professor]. AB - Some personal and professional memories about Mirko Grmek in Switzerland. PMID- 11917916 TI - [Testimonies and letters]. AB - Testimonies from Belgium, Israel, Italy, the Netherlands, and the French province. PMID- 11917918 TI - [A female anatomist of the enlightenment: Marie Marguerite Biheron (1719-1795)]. AB - Since a notice published in 1811 in the Biographie universelle of Michaud, it has been supposed that the famous "Mademoiselle Biheron" of the XVIIIth century, who made artificial anatomies and was especially admired by Diderot, had Marie Catherine as her Christian names. It was a mistake which would invalidate for almost two centuries all the researches on the biography of this anatomist and artist. No one even knew the date and the circumstances of her death. The author, having discovered her true names - Marie Marguerite -, realised many investigations on her and her family. He worked especially on notarian archives, both in Paris, where she was born, and in the north of the old Maine province (present Sarthe department), from which her father, who was a Parisian apothecary, originated. Also he recalls the essential part that this Marie Marguerite Biheron (1719-1795) played in the evolution of the ideas concerning the teaching and medical use of anatomical data. Her contribution in that field was one of those which prepared the important reforms which would take place at the end of the century, at the time of the Convention, conducting in particular to the development of modern surgery. PMID- 11917917 TI - [Development of the surgical school of Lorraine, 1872-1919]. AB - Shortly after the 1870-1871 war between France and Prussia, the Faculty of Medicine of Strasbourg was transferred to Nancy. Few works were dedicated to the surgeons of this new school of Medicine, most of them alsatian at its beginning. Yet the surgeons played an important role in the technical success and certainly management of the Faculty as A. Heydenreich and especially F. Gross, who were the most wellknown Deans in Nancy, during this period. So the new Faculty of Nancy trained many surgeons who took a big place in the Military health Service of the French Army during the 1914-1918 war which ended the first third of the story of this Surgery school. PMID- 11917919 TI - [Gerontology and endocrinology at the beginning of the 20th century: Dr. Voronoff (1866-1951)]. AB - Rejuvenation by monkey testicle transplanting was terribly fashionable in Paris, between years 1925-1930. The manual miracles done by a group of very well-known surgeons, in Paris, (especially Serge Voronoff and Dartigues) rooted in the head of a too credulous public the idea according to it was possible to be made scientifically younger by genital graftings. The "Etude sur la vieillesse et la rajeunissement par la greffe" (1926), written by Voronoff, provided the clinical, histological and iconographical proofs of a such rejuvenation. The paper analyses that was put forward as "proofs", situated in the scientific and cultural context of the time. A possible link between youth, sexual vigour and animality sends back to a mythic history of gerontology and endocrinology, two medical fields which however became scientific specialties in the beginning of the twentieth century. PMID- 11917920 TI - From the Centers for Disease Control and Prevention. Tularemia--United States, 1990-2000. PMID- 11917921 TI - From the Centers for Disease Control and Prevention. Congenital malaria as a results of Plasmodium malariae--North Carolina, 2000. PMID- 11917922 TI - From the Centers for Disease Control and Prevention. Use of assisted reproductive technology--United States, 1996 and 1998. PMID- 11917923 TI - JAMA patient page. Delivering a healthy infant. PMID- 11917924 TI - Houseboat-associated carbon monoxide poisonings on Lake Powell--Arizona and Utah, 2000. AB - During August 2000 at Lake Powell in the Glen Canyon National Recreation Area on the Arizona-Utah border, two brothers died of carbon monoxide (CO) poisoning as they swam near the stern of a houseboat while the onboard gasoline-powered generator was operating. As a result of these deaths, an investigation was initiated by the U.S. National Park Service (NPS) with assistance from the U.S. Department of the Interior, CDC's National Institute for Occupational Safety and Health, and the U.S. Coast Guard. In addition to investigating the deaths of the two brothers, the multiagency team evaluated visitor and worker boat-related CO exposures at Lake Powell. The study identified nine boat-related fatal CO poisonings since 1994 and approximately 100 nonfatal poisonings since 1990. This report describes the preliminary results of an ongoing investigation of watercraft-related CO poisonings on Lake Powell. PMID- 11917925 TI - Unpowered scooter-related injuries--United States, 1998-2000. AB - Injuries associated with unpowered scooters have increased dramatically since May 2000. These scooters are a new version of the foot-propelled scooters first popular during the 1950s. Most scooters are made of lightweight aluminum with small, low-friction wheels similar to those on in-line skates. They weigh <10 pounds and fold for easy portability and storage. Up to 5 million scooters are expected to be sold in 2000, an increase from virtually zero last year (Consumer Product Safety Commission [CPSC], unpublished data, 2000). This report summarizes the results of a descriptive analysis of scooter-related injuries during the past 34 months and provides recommendations to reduce these injuries. PMID- 11917926 TI - Human rabies--California, Georgia, Minnesota, New York, and Wisconsin, 2000. AB - On September 20, October 9, 10, 25, and November 1, 2000, persons who resided in California, New York, Georgia, Minnesota, and Wisconsin, respectively, died of rabies. This report summarizes the case investigations. PMID- 11917927 TI - Human rabies--Quebec, Canada, 2000. AB - On September 22, 2000, a 9-year-old boy awoke with a fever and complained of pain in his upper left arm. The pain persisted, and he developed insomnia and tremors in his arm and hand. He was admitted to a local hospital on September 27. That evening, he had mild dysphagia, pruritus of his upper chest and back, and a transient macular rash. On September 28, he developed tremors and myoclonic jerks in both arms, had become agitated, and had hydrophobia, aerophobia, dysarthria, and visual hallucinations. The next day hypersalivation was observed and the tremors and myoclonus had spread to his lower extremities. He became very anxious, indicated that he was suffocating, and underwent endotracheal intubation. A diagnosis of rabies was considered and he was transferred to a children's hospital. Laboratory findings were normal except a mildly elevated cerebral spinal fluid protein. An electroencephalogram form indicated no epileptiform activity. Head magnetic resonance imaging was normal. On September 29, the results of the rabies tests were positive, and rabies immune globulin and vaccine were administered to the patient. His neurologic and hemodynamic status deteriorated, and he died on October 6. PMID- 11917928 TI - Recommendations from meeting on strategies for improving global measles control, May 11-12, 2000. AB - During May 11-12, 2000, World Health Organization (WHO), United Nations Children's Fund (UNICEF), and CDC co-sponsored a technical working group meeting to review the status of global measles control and regional elimination efforts and to formulate recommendations to accelerate control activities, particularly in countries and regions with a high disease burden. PMID- 11917929 TI - Recent developments in health insurance, life insurance, and disability insurance case law. PMID- 11917930 TI - Recent developments in medicine and law. AB - This article addresses important developments in the field of medicine and law during the period of September 2000 through August 2001. It does not discuss every reported case or piece of legislation but surveys some of the more significant developments in the medicine and law arena. PMID- 11917931 TI - OPRR issues final findings in its investigation of university. PMID- 11917932 TI - State court says informed consent procedures were unacceptable. PMID- 11917933 TI - Institutional review boards and stem cell research. PMID- 11917934 TI - IRBs and privacy of health research data. PMID- 11917935 TI - New requirements for vulnerable subjects and IRB membership. PMID- 11917936 TI - New requirements for "responsible conduct of research" include human subjects area. PMID- 11917937 TI - Violation of compliance rules led to university terminations. PMID- 11917938 TI - Subject's parents claim university was not responsive to their requests. PMID- 11917939 TI - Subjects must have access to treatment and research records. PMID- 11917940 TI - Research regulations on subjects' "capacity to give consent" were unconstitutional. PMID- 11917941 TI - Plasma bcl-2 and nitric oxide: possible prognostic role in patients with metastatic breast cancer. AB - Interest in translational studies on breast cancer is presently devoted to identifying biological predictors of disease prognosis and response to treatment. In this study, we determined the plasma levels of bcl-2 and nitric oxide in 45 patients with metastatic breast cancer using an ELISA technique and correlated them with clinical and biological factors that may affect the outcome of disease. The results were as follows. The mean level of bcl-2 was 278.44 +/- 383.2 U/L compared with 64.42 +/- 14.4 U/L (p = 0.007) for controls. Levels of bcl-2 were higher in patients less than 50 yr old, premenopausals., GIII tumors, positive nodes, ER positive tumors (p = 0.6, 0.5, 0.9, 0.4, and 0.005, respectively). The site of metastatic disease and the number of metastatic sites did not show statistically significant influences over bcl-2 levels. Furthermore, there was a trend, although not significant, toward improvement of survival in patients with higher levels of bcl-2. The mean level of the nitric oxide (NO) was 297.12 +/- 220.54 microM compared with 13.91 +/- 1.1 microM for controls (p = 0.003). The levels were higher in patients over 50 yr, postmenopausal patients, those with visceral deposits, grade III tumors, positive lymph nodes, and those with disease free survival of less than 6 mo following primary treatment (p = 0.1, 0.2, 0.1, 0.09, 0.4, and 0.08 respectively). Furthermore, there was no correlation between NO levels and survival (r = 0.002). This study demonstrates a potential role for NO and bcl-2 as prognostic factors in patients with metastatic breast cancer. However, larger studies with more patients together with a comparison of serum levels (ELISA) and tissue levels (MOAb) are still required. PMID- 11917942 TI - Identification of a gene frequently mutated in prostate tumors. AB - Although prostate cancer is the second leading cause of cancer death for men in the United States, the genetics of tumor development are poorly understood. Several expressed sequence tagged genes (ESTs) that are expressed predominantly in the prostate have recently been identified, although their role in the development and maintenance of the prostate is unknown. Here, we demonstrate that the gene identified as UNIGENE cluster Hs. 104215, which codes for a message found predominantly in the prostate, may be important in tumor development. We name this gene PCan1 for Prostate Cancer gene 1. Northern blot experiments were performed using RNA isolated from tumor-derived cell lines and human prostate to determine the expression pattern of the gene. DNA sequencing was used to identify mutations that occurred in tumor tissue. By Northern blot analysis, this gene product was not detectable in LNCaP, DU 145, or PC-3 prostate cancer cell lines, although it was readily observed in RNA isolated from total prostate and from dissected central and peripheral regions of prostate. Sequence analysis of genomic DNA from LNCaP, DU 145, or PC-3 cells demonstrated a G/A polymorphism at position 193. Analysis of matched tumor-derived DNA and blood-derived DNA samples from 11 of 13 patients who had undergone a radical prostatectomy and who were homozygous for A in blood-derived DNA demonstrated mutation of position 193 in matched tumor samples resulting in G/A polymorphism. Sixteen additional patient samples were G/A polymorphic in both blood-derived DNA and tumor-derived DNA and two samples were GG in both blood-derived and tumor-derived DNA. Our results suggest that this gene may be a hot spot for mutation in prostate cancer, especially because our radiation hybrid mapping located this gene within a region identified in linkage mapping studies of affected families with prostate cancer. Loss of heterozygosity in prostate tumors has also been reported at the location of PCan1. Further studies to determine the functional role of this candidate tumor suppressor gene are warranted. PMID- 11917943 TI - Vinblastin-carboplatin for metastatic cutaneous melanoma as first-line chemotherapy and in dacarbazine failures: a single-center study. AB - First-line treatments of metastatic melanoma are usually decarbazine (DTIC) and/or alpha-interferon based, with response rates in the range of at most 20 30%. In this study, initiated, in fact, by a temporary DTIC shortage in the country, we have assessed the efficacy and toxicity of a vinblastine-carboplatin regimen for metastatic melanoma. The regimen was subsequently applied in two cohorts of patients: a chemotherapy-naive one and in DTIC failures (because the regimen was claimed non-cross-resistant). The regimen contained 6 mg/m2 vinblastine on d 1 and 450 mg/m2 carboplatin on d 1 for 3 wk. In the chemotherapy naive cohort, 50 patients were included, 29 males and 21 females, median age 54 yr (range: 33-68), performance status 0+1 for 26 patients and 2+3 for 24 patients. Forty-eight patients were evaluable for activity. The response was the following: complete response (CR), 1/48 (2%); partial response (PR), 13/48 (27%); stable disease (SD), 20/48 (42%); progressive disease (PD), 14/48 (29%). The overall response rate was 14/48 (29%). The median response duration was 7 mo (range: 3-14); the median time to progression was 4 mo (range: 2-14). Toxicity included granulocytopenia and thrombocytopenia grade IV in 3/50 patients and nausea grade II in 8/50 patients. In the DTIC-failures cohort, 58 patients were included, 38 males and 20 females, median age 51 yr (range: 20-65), performance status 0+1 for 25 patients and 2+3 for 33 patients. All 58 patients were evaluable for activity. The response was the following: CR 3/58 (5%), PR 4/58 (7%), SD 10/58 (17%), PD 41/58 (71%). The overall response rate was 7/58 (12%). The median response duration was 11 mo (range: 3-24); the median time to progression was 4 mo (range: 2-24). Toxicities included granulocytopenia grade IV in 4/58 patients and nausea grade II in 4/58 patients. Thus, despite the fact that the regimen achieved a response rate comparable to DTIC in a first-line setting, the lack of cross-resistance did not prevent it from being of limited activity in DTIC failures, although, even in this group, several long-lasting responses and stabilizations were noted. PMID- 11917944 TI - Kidney cancer: the Cytokine Working Group experience (1986-2001): part I. IL-2 based clinical trials. AB - The Cytokine Working Group (CWG) was initially established in 1986 as the Extramural IL-2/LAK Working Group. With funding from the National Cancer Institute (NCI), the CWG was mandated to confirming data regarding the efficacy of the high-dose interleukin-2 (IL2)/lymphokine-activated killer cell (LAK cell) regimen piloted at the NCI in the treatment of renal cell cancer. Since those initial studies, the CWG has conducted a series of clinical trials, often with correlative immunologic investigations, to evaluate combination immunotherapy in attempts to enhance the efficacy of IL-2 or to reduce toxicity. Subsequently, the CWG conducted trials to demonstrate the activity of lower-dose outpatient combination cytokine regimens to help determine their role in the armamentarium of treatment for metastatic renal cell cancer. This has culminated in a phase III randomized trial comparing the activity of high-dose IL-2 with the activity of outpatient IL-2 plus interferon-alpha. The CWG also has honed the management of both high-dose IL-2 and outpatient IL-2 regimens to make these safer in the hands of experienced clinicians. In addition, the CWG has produced a series of carefully conducted clinical trials of new cytokines, again attempting to define their clinical efficacy as anticancer agents. These include studies of IL-4, IL 6, and IL-12. Currently, the CWG is conducting studies with new approaches to IL 2 therapy, as well as planning trials with new agents for treatment of renal cell cancer. This review describes these efforts conducted over the past 15 yr. PMID- 11917946 TI - Extragonadal seminoma after renal transplantation and immunosuppression; treatment in the presence of renal dysfunction: a case report and literature review. AB - A 37-yr-old man who had undergone renal transplantation for end-stage renal failure presented with a large right pelvic mass obstructing the transplanted kidney. Initially, this was diagnosed as an anaplastic tumor while he had been on immunosuppressive treatment for kidney allograft rejection after transplantation. Despite difficulties of classic histopathology to reveal the origin of his tumor, FISH analysis revealed the presence of chromosome 12p abnormalities, strongly indicative of a germ-cell tumor-more likely seminoma-with extragonadal presentation. Because of renal dysfunction, he was treated with carboplatin (dose adjusted according to renal clearance) and etoposide, and when he experienced a rather atypical progression with bone metastases, he was treated with single agent paclitaxel, and died almost 13 mo after initial presentation. The case adds further to the existing small list of seminoma/GCTs developing in transplant recipients, points to the unusual presentation patterns and diagnostic histopathology challenges, and presents the difficulty in therapeutic options, as a result of frequent renal dysfunction and intercurrent immunosuppressive therapy. All of these issues together with an extensive literature review are discussed in detail. PMID- 11917945 TI - Kidney cancer: the Cytokine Working Group experience (1986-2001): part II. Management of IL-2 toxicity and studies with other cytokines. AB - The Cytokine Working Group (CWG) was initially established in 1986 as the Extramural IL-2/LAK Working Group. With funding from the National Cancer Institute (NCI), the CWG was mandated to confirming data regarding the efficacy of the high-dose interleukin-2 (IL2)/lymphokine-activated killer cell (LAK cell) regimen piloted at the NCI in the treatment of renal cell cancer. Since those initial studies, the CWG has conducted a series of clinical trials, often with correlative immunologic investigations, to evaluate combination immunotherapy in attempts to enhance the efficacy of IL-2 or to reduce toxicity. Subsequently, the CWG conducted trials to demonstrate the activity of lower-dose outpatient combination cytokine regimens to help determine their role in the armamentarium of treatment for metastatic renal cell cancer. This has culminated in a phase III randomized trial comparing the activity of high-dose IL-2 with the activity of outpatient IL-2 plus interferon-alpha. The CWG also has honed the management of both high-dose IL-2 and outpatient IL-2 regimens to make these safer in the hands of experienced clinicians. In addition, the CWG has produced a series of carefully conducted clinical trials of new cytokines, again attempting to define their clinical efficacy as anticancer agents. These include studies of IL-4, IL 6, and IL-12. Currently, the CWG is conducting studies with new approaches to IL 2 therapy, as well as planning trials with new agents for treatment of renal cell cancer. This review describes these efforts conducted over the past 15 yr. PMID- 11917947 TI - Busulfan-induced loss of Ph chromosome in chronic myeloid leukemia. AB - We report a 44-yr-old female with Philadelphia chromosome positive chronic myeloid leukemia who was initially treated with busulfan, and clinical remission has been achieved. After 4 yr, low-dose busulfan therapy was started again and induced bone marrow aplasia. The patient spontaneously has recovered from aplasia, and complete cytogenetic remission with loss of Ph+ chromosome in bone marrow has been achieved. However, reverse-transcription-polymerase chain reaction analysis showed presence of the bcr-abl transcript in bone marrow. PMID- 11917948 TI - GM-CSF at relatively high topic concentrations can significantly enhance the healing of surgically induced chronic wounds after radiotherapy. AB - Combinations of radiotherapy and surgery are often used in local cancer treatments. Preoperative radiotherapy may delay wound healing after surgery. Chronic wounds are debilitating conditions that require frequent medical attention. Two patients suffering from chronic and slowly healing wounds post surgery and preoperative radiotherapy are described. A significant acceleration of the healing by local injections with GM-CSF was demonstrated. PMID- 11917949 TI - Ca 19-9 in the monitoring of colorectal cancer after surgery. PMID- 11917950 TI - Fasciocutaneous free flaps for hypopharyngeal reconstruction. AB - Considerable controversy persists regarding the optimal technique for hypopharyngeal reconstruction. The ideal procedure should provide low mortality and morbidity, short hospitalization, a high success rate, few complications, and the greatest potential for neopharyngeal speech and deglutition. In this study, a variety of fasciocutaneous free flaps were used for reconstruction of the hypopharynx. Over a two-year period, fasciocutaneous flaps were used for reconstruction of pharyngoesophageal segments following total laryngopharyngectomies in 16 patients at The National Cancer Institute, Cairo University, Cairo, Egypt. Flap survival, recipient vessels used, and complications were examined. The ultimate functional and cosmetic outcomes of free flaps were compared. Of the 16 patients included in this study, nine were males, and seven were females. Free flaps used for reconstruction included the radial forearm (8), lateral arm (2), anterolateral thigh (3), and posterolateral thigh (3) flaps. Free flaps were successful in 15 patients. One patient had total flap loss. Salvage surgery was successful for one flap that developed venous congestion. Eleven patients received adjuvant radiation therapy. The commonly used recipient vessels were the small arteries of the neck and the external jugular vein. Five patients developed minor pharyngocutaneous fistulas that healed spontaneously. Six patients developed anastomotic line stricture. Donor site morbidity was more significant with the radial forearm flap, compared to other flaps. Fasciocutaneous free flaps have a definite place in pharyngoesophageal reconstruction. The flap should be selected with reference to the type of the defect and patient obesity; however, donor-site morbidity should also be considered. PMID- 11917951 TI - Sequential microsurgical flap reconstruction following purpura fulminans during infancy and childhood. AB - Purpura fulminans is a potentially lethal complication of meningococcal septicemia, characterized by progressive hemorrhagic skin lesions, which can result in extensive necrosis and mummification of all the extremities. With improving survival rates in infancy and childhood, plastic surgeons are challenged more often to provide sufficient and stable soft-tissue coverage. Usually, conservative methods, such as skin grafting or amputation, are favored by many pediatric surgeons, since further specialized departments and training are not required. Often secondary reconstructive procedures to improve soft tissue coverage have to be performed to achieve proper prosthetic fitting. Microsurgical techniques are used only in selected cases, after failure of other procedures for defect coverage. In two cases of post-acute purpura fulminans, two free flaps and three microsurgically dissected flaps were used as primary measures for defect coverage and preservation of stump length. Despite the presence of vasculitis, all flaps survived. In a third case, secondary reconstructive measures had to be performed 1 year after purpura fulminans due to insufficient soft-tissue coverage after lower leg amputation. This patient also had contractures on both hands and no grip function after complete finger loss. Several microsurgical procedures were performed to improve grip function and soft tissue coverage. The primary use of microsurgical techniques prevents lengthy secondary reconstructive measures. PMID- 11917952 TI - Sense and sensibility: breast reconstruction with innervated TRAM flaps. AB - During a 25-month period, ten patients underwent post-oncologic breast reconstruction with innervated TRAM flaps. All patients were noted to have a more rapid and full return of sensation by the modalities tested than has been reported among patients with non-innervated TRAM-flap reconstructions. No complications relating to the microneural surgery were noted. Three patients experienced complications common to flaps in general: specifically, these were skin-flap necrosis in the cutaneous portion of one flap, skin necrosis of the infraumbilical skin flap at the closure site, and abdominal bulging at the donor area. Given the minimal increase in operating time required for the completion of the innervated TRAM-flap reconstruction and the obvious benefits in terms of postoperative sensation, this seems potentially to be an important modification to what is already a well-established technique. PMID- 11917953 TI - Combined latissimus dorsi-thoracodorsal artery perforator free flap: the "razor flap". AB - Elevation of the skin island overlying the latissimus dorsi muscle, dissecting the dominant perforating vessel, permits independent positioning of the skin island in relation to the muscle. This combination of the thoracodorsal artery perforator (TAP) and the latissimus dorsi (LD) muscle flap expands the surface of the flap without increasing donor morbidity. PMID- 11917954 TI - Direct closure of radial forearm free-flap donor sites by double-opposing rhomboid transposition flaps: case report. AB - A basic disadvantage of the radial forearm flap is the removal of skin from a functionally important and cosmetically exposed region. The donor site is conventionally repaired by skin grafting. However, this method is associated with complications of graft take and a poor aesthetic appearance. In this report, the authors describe a simple technique for direct closure of a distal forearm flap donor defect, using double-opposing rhomboid transposition flaps. This method is based on the existence of an oblique skin laxity in the distal forearm from the ulnar to the radial side, allowing a double-opposing local flap design. In a 32 year-old female patient, a 6- X 4-cm longitudinally-oriented elliptical skin defect of a radial forearm free flap was succesfully closed directly with the described technique without any complications. This method is a useful alternative for primary closure of small- to medium-sized distal forearm defects. PMID- 11917955 TI - Late effects of TNF-alpha-induced inflammation on the microcirculation of cremaster muscle flaps under intravital microscopy. AB - In order to understand the microcirculatory changes and regulatory mechanisms governing passage of neutrophils from the vascular bed to the interstitial tissue during ischemia/reperfusion (I/R) injury, a key component of this injury, tumor necrosis factor-alpha (TNF-alpha)-induced inflammation was analyzed. Twenty-four Sprague-Dawley rats were divided into four groups, containing six animals in each. The effect of TNF-alpha-induced inflammation was studied at two different time points, early sequential and late. In the early-effect Groups 1 and 2, animals were given TNF-alpha and vehicle, respectively. Microcirculatory changes were recorded for 6 hr continuously. In the late-effect Groups 3 and 4, following TNF-alpha injection and vehicle, microcirculatory changes were measured 16 hr later. In the early-effect groups, the number of rolling and adhering leukocytes was increased immediately following TNF-alpha injection and remained elevated for the first 3 hr (p<0.05). The number of transmigrated leukocytes remained significantly increased throughout the first 6 hr (p<0.05) and returned to normal at 16 hr. In delayed-effect groups, a second peak in the number of rolling leukocytes was noted at 16 hr (p<0.05). The numbers of rolling and adhering lymphocytes, although remained at the baseline for the first 6 hr, was increased 2- and 1.5-fold at 16 hr, respectively (p<0.05). The number of perfused capillaries gradually decreased over time in the TNF-alpha-induced inflammation groups. A vasodilatory response was noted at the third and fourth order arterioles within the first 3 hr of measurement (p<0.05), but returned to normal afterward. The detrimental effects of TNF-alpha-induced inflammation during I/R injury could be prolonged up to 16 hr at the microcirculatory level of the muscle flaps. PMID- 11917957 TI - Effect of acute ischemic preconditioning on blood-flow response in the epigastric pedicled rat flap. AB - Research in the field of microvascular surgery has shown that ischemic preconditioning (repeated brief episodes of feeding artery occlusions followed by reperfusion) improves flap survival. The authors used a custom-designed clamping method and laser Doppler flowmetry to investigate changes in blood flow (BF) responses, either with acute ischemic preconditioning or without it. The animal model used was the partially elevated epigastric flap of adult Sprague-Dawley rats. In the preconditioned group (n=12), the flaps underwent preconditioning with three cycles of 10-min of feeding artery clamping, followed by 10 min of reperfusion, for a total preconditioning period of 1 hr. In the control group (n=12), the flaps were perfused without clamping for 1 hr. All the flaps underwent occlusion of the feeding artery for 15, 30, 60, 120 and 180 sec to observe the changes in the BF responses, 60 min and 150 min after flap elevation. To compare the responses between preconditoned and control groups the BF responses were analyzed during the overshoot period (i.e., BF being above the baseline after different feeding artery occlusion periods). Statistical analysis of the responses showed that the magnitude of increase in BF after clamp release (p<0.001), the duration of overshoot (p=0.014), and the amplitude of overshoot after clamp release (p=0.002) were statistically significantly greater in the preconditioned group than in the control group. The results suggest that vessels and their responses to change in perfusion pressure are involved in the multifactorial process of the ischemia-protective effect caused by acute ischemic preconditioning. As far as is known, this is the first report showing changes in flap vascular responses after acute ischemic preconditioning. PMID- 11917956 TI - Effect of a nitric oxide donor on microcirculation of acutely denervated skeletal muscle during reperfusion. AB - The authors have shown that exogenous nitric oxide (NO) protects innervated skeletal muscle against reperfusion injury. This study further evaluated the effects of exogenous NO donor on denervated skeletal muscle. Forty-eight denervated rat cremaster muscles underwent 3 hr of ischemia, followed by 90 min of reperfusion, and received systemic infusion of 100 nmol/min s-nitroso-n acetylcysteine (SNAC) or an equal amount of phosphate-buffered saline (PBS). Results showed that the average diameter in 10 to 20 microm arterioles was between 107 percent and 123 percent of baseline in the SNAC group, and between 55 percent and 84 percent in the PBS group during 90 min of reperfusion. These values in 21 to 40 microm and 41 to 70 microm arteries were between 100 percent and 110 percent in the SNAC group, and between 70 percent and 90 percent in the PBS group from 20 to 90 min of reperfusion. Compared to the PBS group, the SNAC group had a statistically significantly greater vessel diameter in both 10 to 20 microm (p<0.001) and 21 to 40 microm arterioles (p<0.01) during 90 min of reperfusion, and in 41 to 70 microm arteries (p<0.02) from 20 to 90 min of reperfusion. The overall blood flow of the muscle in the SNAC group increased from 37 percent of baseline at 10 min to 108 percent at 40 min of reperfusion, and remained above baseline thereafter. In contrast, this value in the PBS group was only between 27 percent and 68 percent of baseline during 90 min of reperfusion. The blood flow was statistically significantly (p<0.03) greater in the SNAC group than in the PBS group from 40 to 90 min of reperfusion. Among the conclusions were: (1) NO donor SNAC improves the microcirculation of denervated skeletal muscle during early reperfusion; and (2) this protection against reperfusion injury is independent of innervation in skeletal muscle. PMID- 11917958 TI - Rare variant of the intrasoleus musculocutaneous perforator: clinical considerations in raising a free peroneal osteocutaneous flap. PMID- 11917959 TI - Outcomes of surgical treatment of brachial plexus injuries using nerve grafting and nerve transfers. AB - Between 1993 and 1998, 32 male patients with brachial plexus injuries were surgically treated. Eighteen interfascicular grafting and 71 extraplexal neurotization procedures were performed separately or in combination. Donor nerves were the intercostals, spinal accessory, phrenic, contralateral C7, and cervical plexus, in order of frequency. Patients were followed for a minimum of 24 (average, 35) months. Biceps function was best following grafting the musculocutaneous nerve itself, or neurotization with the phrenic nerve (100 percent grade 4), followed by neurotization with the intercostals (89.5 percent grade 3 or more) and last, grafting the C5 root or upper trunk (grade 3 in one of three patients). Phrenic to suprascapular neurotization produced the best results of shoulder abduction (40 to 90 degrees), followed by combined neurotization of the spinal accessory to suprascapular and phrenic to axillary (20 to 90 degrees). Sensory recovery over the lateral forearm and palm varied from S2 to S3+, according to the method of reconstruction. PMID- 11917960 TI - 11 September 2001--and after. PMID- 11917962 TI - Globalization, migration and health. AB - The term 'globalization' describes the integration of economic systems through improved communication, but it also represents increased insecurity for those with few resources--particularly refugees. This article examines why people migrate, their numbers, constraints on their movement and their particular health care needs. Immigrants have much to contribute to their recipient countries, but at some loss to their homelands. Both economically and morally, more liberal immigration policies would be beneficial. Policies towards asylum seekers should not be more restrictive in the aftermath of 11 September 2001 and detention should be the exception rather than the rule. Globalization should be managed so as to improve people's lives throughout the world. PMID- 11917961 TI - Adolescents and political conflict in Northern Ireland. PMID- 11917963 TI - Psychosocial response to disaster: the attacks on the Stark and the Cole. AB - The terrorist attack on the USS Cole on 12 October 2000 was remarkably similar to the 1987 attack on the USS Stark. This article discusses the psychosocial consequences of the attacks on the families and crews of the ships and the community response of the Navy to the attacks, particularly that of the Navy Family Service Centers. The impact of the attacks is compared to the impact of natural and man-made disasters on communities while the impact on the crew is examined in light of combat psychiatry and post-traumatic stress disorder (PTSD). Events such as these are very likely to produce PTSD despite early intervention efforts. Following the attack on the Stark greater attention was given to the grief of family members than to the trauma of the crew, while the crew of the Cole has received longer-term psychiatric assistance than in previous similar episodes. PMID- 11917964 TI - Setting the scene: the new conflict environment and contemporary challenges for interventionists. AB - This article examines the characteristics of contemporary conflict and humanitarian environments and looks at the reasons that underpin their evolution. It studies the role of globalization, international economies, weapons proliferation, warlords, individual security guarantees and urbanization. The role of the international community is discussed and the main responsibilities and actors are defined. Future challenges for multinational military forces are examined and an overview given of the role of non-governmental organizations (NGOs), United Nations (UN) agencies, bilateral and multilateral donors, commercial companies and host governments. Lastly, recommendations are given on how the international community can close existing gaps that impede operational effectiveness, whilst still providing integrated responses to global needs. PMID- 11917965 TI - The case of Serbia/Yugoslavia: an analysis through spiral dynamics. AB - Yugoslavia and later Serbia have puzzled the world since the early 1990s. This article presents a study based on the principles of spiral dynamics that were used to analyze the transformation of South Africa. According to spiral dynamics, Yugoslavia under the joint influence of nationalists and retro-socialists regressed to the egocentric and exploitative level of psychosocial existence and, in spite of recent political changes, has maintained the same centre of gravity. Although its political structure has changed, nationalist and retro-socialist values remain dominant in the cultural domain and in the domain of self. Yugoslavia/Serbia desperately needs the new dynamic process of development. Priorities are the establishment of a hierarchy of authority, self-confrontation and development of widely based enthusiasm. These could lead Serbia to become responsible and mature. PMID- 11917966 TI - Pipelines and poppies. PMID- 11917968 TI - International Physicians for the Prevention of Nuclear War (IPPNW) International Medical Conference on Small Arms, Gun Violence and Injury: A call for action to reduce small arms deaths and injuries. PMID- 11917967 TI - Prevention of war is a precondition for prevention of nuclear war. PMID- 11917969 TI - Humour as a strategy in war. PMID- 11917970 TI - Thrombolysis for acute ischemic stroke: the problem of consent. AB - The physician who decides to administer a thrombolytic therapy to a patient with acute stroke, has to consider both the results of randomized controlled trials, a complex matter itself, and problems related to consent, with its deontological and legal aspects, in the difficult contest of the emergency situations and acute stroke care. Although the analysis of studies on thrombolysis in medical literature has been ample and may be redundant, the latter aspect, i.e. that of how to cope with the problem of consent, has never been treated adequately. The problem of consent is particularly important and delicate in Italy and in most countries of the European Union, where thrombolytic therapy can be used only off label or within experimental studies. This document aims to provide the tools needed to overcome the gap between the generic indication for thrombolytic therapy in acute stroke, coming from clinical studies, and its effective use. PMID- 11917971 TI - Transcranial Doppler in the diagnosis of cardiac patent foramen ovale. AB - Right-to-left shunt due to patent foramen ovale is a well-established risk factor for ischemic stroke, especially in young subjects and in patients with cryptogenic stroke. We report a detailed method for the diagnosis of a right-to left shunt by means of transcranial Doppler as established at a recent consensus conference, and review the literature on the indications for the test. PMID- 11917972 TI - Experience with a questionnaire administered by emergency medical service for pre hospital identification of patients with acute stroke. AB - We prospectively verified whether an ad-hoc questionnaire administered by phone supports pre-hospital suspicion of stroke in order to reduce the time before care is given. From June 1996 to May 1997, physicians of the Emergency Medical Service in the area of Bergamo, Italy asked all people calling for a patient with symptoms and signs suggesting a cerebral vascular injury to immediately answer some questions on common symptoms and signs of stroke. The medical records of the patients hospitalized at Ospedali Riuniti of Bergamo were reviewed at the end of the study by a single neurologist, skilled in stroke management and blinded to the questionnaires. Sensitivity and specificity, in addition to positive and negative predictive values, of single questions versus final diagnosis were assessed. Logistic regression analysis was also performed to identify those questions useful to suspect strokes. We collected 143 valid questionnaires, related to 63 men and 80 women, aged 34-99 years (mean, 71.8 years). The question concerning headache had the lowest sensitivity and specificity, respectively 57.1% and 36.5%, and the question concerning leg palsy had the highest sensitivity and specificity, respectively 82.0% and 52.4%. Multivariate analysis identified questions on facial and leg palsy as independent predictors of a final diagnosis of stroke. A few questions on motor deficits proposed by emergency medical service operators may be useful in the pre-hospital identification of stroke patients. Concordance of any questions versus final diagnosis of stroke was, however, far to be satisfying. Thus, our experience supports the need for an educational program to improve the efficiency of a pre-hospital diagnosis of stroke. PMID- 11917974 TI - Evolution of upper limb function in children with congenital hemiplegia. AB - Hand function deficits in hemiplegic children are a major cause of disability, but there is a lack of appropriate instruments for evaluating the evolution of this deficit over time and for verifying the efficacy of its treatment. We evaluated changes in upper limb function in relation to age and the course of individual rehabilitation treatment in 20 children (13 males and 7 females) who were first seen within the first four years of life and subsequently followed until a mean age of 13 years and four months (range, 11-17 years) in accordance with a diagnostic/rehabilitation program initiated in our division in 1989. All of the children were treated by us; those whose paretic upper limb functioned well were not treated in any specific or directed manner. The protocol involved a qualitative evaluation of the spontaneous use of the paretic hand and a quantitative evaluation of grip. Analysis of the results revealed an age-related global improvement over time, occurring within the first five years of life and more pronounced in terms of grip than spontaneous use. This finding makes our protocol more specific than those currently used because it more reliably establishes the real capacity to use the paretic hand in different situations of everyday life. The most important changes concerned the children with more impaired functional capacity, whereas the children who presented with good functional skill retained this capacity over time, thus confirming the initial decision not to treat them. PMID- 11917973 TI - Health-related quality of life in patients with myasthenia gravis and the relationship between patient-oriented assessment and conventional measurements. AB - We assessed the health-related quality of life (HRQoL) of patients with myasthenia gravis (MG) and correlated it with the physician's measurements of MG. Patients with MG were evaluated by means of (1) self-administered questionnaires, (2) clinical examination, (3) Osserman classification, (4) anti-AChR antibody, and (5) neurophysiology. Relationships between patient-oriented assessment and conventional clinical-neurophysiological and serological findings were evaluated. A total of 46 patients, inpatients and outpatients (mean age 50.7 years, range 11 77 years, 17 males, 29 females) with MG diagnosis were studied. The Osserman scale and clinical examination findings were significantly related to the physical aspects of HRQoL. Mental aspects of the quality of life were not progressively involved as muscle deficit progressed, but even in a mild clinical picture, the mental aspects were deteriorated. Patient-oriented measures proved that the patient's quality of life was impaired especially with regard to physical aspects. Our data demonstrated that clinical measurements are related to the HRQoL. The results may be useful in developing a disease-specific patient oriented tool. PMID- 11917975 TI - Developmental patterns of verbal and visuospatial spans. AB - This study presents developmental data for verbal and spatial memory tasks: Corsi's block-tapping test and Luria's verbal learning test. Norms have been collected from 275 primary and early secondary school children aged from 5 years, 4 months to 13 years, 6 months. Our results confirm a slow and constant improvement in performances over time, and the advantage of about 1.5 items of the verbal span over the spatial span supports the existence of developmental differences between separate memory systems. No significant sex difference was found even if a slight trend in verbal span favouring female subjects is present. PMID- 11917976 TI - Life-threatening intracranial hypotension after diagnostic lumbar puncture. AB - Intracranial hypotension syndrome as a complication of diagnostic lumbar puncture is a rarely observed entity. Intracranial hypotension syndrome is characterized by postural headache, neck pain/stiffness, blurred vision, nausea, vomiting, clouding of consciousness, dizziness and vertigo. The majority of cases resolve spontaneously with conservative treatment. Rarely, epidural blood patch is required. We report a 41-year-old man with multiple sclerosis, who developed intracranial hypotension syndrome after diagnostic lumbar puncture and who did not respond to conservative treatment. A subdural hematoma was subsequently found, when the patient showed considerable worsening of clinical conditions with life-threatening symptoms. Surgical evacuation of the subdural hematoma was not sufficient to improve significantly the patient's conditions, while complete symptoms remission was achieved 12 hours after epidural blood patch. We stress the need for epidural blood patch in any case of post-diagnostic lumbar puncture postural headache which does not resolve with conservative therapy. PMID- 11917977 TI - Paroxysmal dystonia with thalamic lesion in multiple sclerosis. AB - Paroxysmal dystonia (PD) is a usually painful, unilateral dystonic posture, precipitated by voluntary movement, tactile stimulation, startling noise or hyperventilation. We describe two cases of paroxysmal dystonia in multiple sclerosis, both with a critically localized lesion in the thalamus, contralateral to the paroxysmal symptoms. Only one other case of paroxysmal dystonia with a demyelinated lesion of the thalamus has been reported previously. PMID- 11917978 TI - Headache as a manifestation of fatal myocardial infarction. AB - Cardiac ischemia typically causes chest pain, variously radiating elsewhere. Convergence of cardiac nerve fibers on central pathways receiving somatic afferents from the head is likely to be responsible for the perception of cardiac ischemic pain as headache. A 47-year-old man was admitted to the emergency room of our hospital with a 2-hour history of occipital headache. Routine electrocardiography revealed monophasic ST-segment elevation in leads I and aVL and ST-segment depression in leads II, III, and aVF. During recording of lead VI, ventricular fibrillation occurred. Advanced life support was started immediately but failed to restore rhythm and cardiac function. Autopsy showed two-vessel disease with a ruptured plaque and total thrombotic occlusion of the proximal left anterior descending artery and 80% stenosis of the right coronary artery. In this patient, headache was the only symptom of myocardial ischemia. Anatomic convergence of cardiac nerve fibers on central pathways receiving somatic afferents from the head is likely to be responsible for the perception of cardiac ischemic pain as headache. Owing to the very rare occurrence of headache as a symptom of myocardial ischemia, diagnosis is difficult and requires a high degree of suspicion. PMID- 11917979 TI - Early thrombolysis in stroke due to basilar artery occlusion. AB - Basilar artery occlusion is usually associated with a poor prognosis. Nevertheless, intra-arterial thrombolysis has been shown to improve clinical outcome in selected cases. We report the case of a 29-year-old patient who suffered a severe ischemic stroke due to basilar artery occlusion and who was treated with intra-arterial thrombolysis within 3 h of symptoms onset; treatment was followed by an almost full functional recovery. The young age of the patient, short segment occlusion in the middle tract of the artery, good collateral supply, and early recanalization may account for the favorable prognosis. PMID- 11917980 TI - Lemierre's syndrome complicated by carotid thrombosis. AB - Lemierre's syndrome, also known as postanginal sepsis, is a rare condition that presents as an increasing sore throat due to acute pharyngitis or tonsillitis and progresses to sepsis, due to suppurative thrombophlebitis of the internal jugular vein. We present an atypical case of Lemierre's syndrome complicated by carotid thrombosis. The etiological factors and the diagnostic and therapeutic measures are discussed. PMID- 11917981 TI - Spinal dysraphism in an elderly patient. AB - Spinal dysraphisms are diagnosed more frequently at birth or in infancy. We report a spinal malformation compatible with lipomyeloschisis in an elderly patient presenting with symptoms and signs of myelopathy. Magnetic resonance imaging revealed an intraspinal mass continuous with a subcutaneous lipoma. Three dimensional computed tomography reconstructions better showed the spinal dysraphism; dermal sinus was also evident. Neuroimaging can define the precise diagnosis also in elderly patients presenting with myelopathy and can provide valuable structural details. PMID- 11917982 TI - Baclofen is effective in intractable hiccups induced by brainstem lesions. PMID- 11917983 TI - Post-lumbar puncture intracranial hypotension. PMID- 11917984 TI - Application of the multimedia urban model to compare the fate of SOCs in an urban and forested watershed. AB - A multimedia model has been developed to estimate the dynamics of semivolatile organic compounds (SOCs) in urban areas. The model is based on a Level III fugacity model of Mackay and consists of six compartments: air, surface water, sediment, soil, vegetation, and an organic film that coats impervious surfaces. The model was used to illustrate the effect of impervious surfaces in urban areas by parametrization for downtown, Toronto, Canada, and modification of the same area to simulate forested conditions. With illustrative emissions of PCB homologues to air, the model indicates that most chemicals are lost by advection, with the remainder undergoing air-to-surface (organic film or vegetation) transfer. Under urban conditions chemicals with Log[K(OA)] < 7.5 volatilize from the film into air where they are susceptible to advection and photolytic degradation. Chemicals with Log[K(OA)] > 7.5 are washed off the film to surface waters where they may undergo volatilization, advection, sedimentation, and degradation. Both loss mechanisms from the film increase the overall mobility of SOCs in the urban relative to the forested environment. In forested areas, vegetation more efficiently accumulates gas- and particle-phase SOCs and subsequently transfers them to surface soils, the greatest chemical reservoir, where they are relatively immobile. PMID- 11917985 TI - Evaluating a model of the historical behavior of two hexachlorocyclohexanes in the Baltic Sea environment. AB - The POPCYCLING-Baltic model, a non-steady-state multicompartmental mass balance model of long-term chemical fate in the Baltic Sea environment, is evaluated for its capability to simulate the behavior of alpha- and gamma-hexachlorocyclohexane isomers from 1970 to 2000. The model predicts HCH concentrations in air, seawater, marine sediments, and needles well within an order of magnitude, often within a factor of 2, and reproduces major features of the observed differences in space and time. The spatial and temporal patterns of HCHs in the Baltic Sea environment are found to be controlled mostly by chemical input, both through direct emissions within the drainage basin and through advective inflow from adjacent areas, but variable environmental conditions can significantly modify the spatial distribution patterns. Simulations with different boundary conditions, i.e., variable assumptions concerning advective atmospheric inflow, suggest that sources within the drainage basin alone are not capable of explaining the observed HCH levels and that significant transport into the region must occur. Deviations between predicted and measured HCH concentrations can often be explained by uncertain estimates of usage and advective import, illustrating the usefulness of the model for evaluating the reasonability of emission estimates and boundary conditions. PMID- 11917986 TI - Mass budgets, pathways, and equilibrium states of two hexachlorocyclohexanes in the Baltic Sea environment. AB - The POPCYCLING-Baltic model, a non-steady-state multicompartmental mass balance model of long-term chemical fate in the Baltic Sea environment, is used to derive a quantitative understanding of the behavior of alpha- and gamma hexachlorocyclohexane (HCH) from 1970 to 2000. The atmosphere is found to effectively distribute the HCHs within the Baltic Sea environment and beyond, resulting in relatively uniform concentrations in environmental compartments that do not directly receive emissions. This uniformity is the result of a large-scale redistribution of a relatively small fraction of the emitted HCHs from the agricultural systems in source areas to all other environmental compartments throughout the Baltic Sea region. The major fraction of the HCHs is degraded in the soils receiving the pesticide application. In areas where HCH-containing pesticides are used, HCHs evaporate from soils and water bodies and are advected away in the atmosphere. They are deposited to forests and water bodies when they reach remote regions. This redistribution is driven by the inclination of the HCHs to equalize their chemical potential within the environment, which is illustrated through the use of fugacity fractions. The model is believed to provide useful insight into the complex set of interactions that determine the overall fate of an environmental contaminant but which are inaccessible to measurements. PMID- 11917987 TI - Can green chemistry promote sustainable agriculture? PMID- 11917988 TI - Dissolution of nonuniformly distributed immiscible liquid: intermediate-scale experiments and mathematical modeling. AB - The purpose of this work is to examine the effect of nonuniform distributions of immiscible organic liquid on dissolution behavior, with a specific focus on the condition dependency of dissolution (i.e., mass transfer) rate coefficients associated with applying mathematical models of differing complexities to measured data. Dissolution experiments were conducted using intermediate-scale flow cells packed with sand in which well-characterized zones of residual trichloroethene (TCE) and 1,2-dichloroethane (DCA) saturation were emplaced. A dual-energy gamma radiation system was used for in-situ measurement of NAPL saturation. Aqueous concentrations of TCE and DCA measured in the flow-cell effluent were significantly less than solubility, due primarily to dilution associated with the nonuniform immiscible-liquid distribution and bypass flow effects associated with physical heterogeneity. A quantitative analysis of flow and transport was conducted using a three-dimensional mathematical model wherein immiscible-liquid distribution, permeability variability, and sampling effects were explicitly considered. Independent values for the initial dissolution rate coefficients were obtained from dissolution experiments conducted using homogeneously packed columns. The independent predictions obtained from the model provided good representations of NAPL dissolution behavior and of total TCE/DCA mass removed, signifying model robustness. This indicates that for the complex three-dimensional model, explicit consideration of the larger scale factors that influenced immiscible-liquid dissolution in the flow cells allowed the use of a dissolution rate coefficient that represents only local-scale mass transfer processes. Conversely, the use of simpler models that did not explicitly consider the nonuniform immiscible-liquid distribution required the use of dissolution rate coefficients that are approximately 3 orders of magnitude smaller than the values obtained from the column experiments. The rate coefficients associated with the simpler models represent composite or lumped coefficients that incorporate the effects of the larger scale dissolution processes associated with the nonuniform immiscible-liquid distribution, which are not explicitly represented in the simpler models, as well as local-scale mass transfer. These results demonstrate that local-scale dissolution rate coefficients, such as those obtained from column experiments, can be used in models to successfully predict dissolution and transport of immiscible-liquid constituents at larger scales when the larger scale factors influencing dissolution behavior are explicitly accounted for in the model. PMID- 11917989 TI - Lead analysis by anti-chelate fluorescence polarization immunoassay. AB - Lead concentrations were determined by a fluorescence polarization immunoassay (FPIA) method that uses polyclonal antibodies raised against the lead(II) chelate of ethylenediamine-N,N,N',N'-tetraacetic acid (EDTA). The technique is based on competition for a fixed concentration of antibody binding sites between Pb-EDTA, formed by treating the sample with excess EDTA, and a fixed concentration of a fluorescent analogue of the Pb-EDTA complex. The objective was to correlate results obtained by FPIA with those produced by conventional atomic spectroscopy analysis of soils, solid waste leachates (produced by the Toxicity Characteristic Leachate Procedure; TCLP), airborne dust, and drinking water. Linear regression analysis of FPIA results for 138 soil samples containing 0-3094 ppm Pb(II) by flame atomic absorption spectroscopy and 40 TCLP extracts containing 0-668 ppm Pb(II) by inductively coupled plasma atomic emission spectroscopy produced correlation coefficients (r2) of 0.96 and 0.93, respectively. Pilot studies of mineral acid extracts of airborne dust trapped on fiberglass filters and of two sources of drinking water demonstrated the feasibility of also measuring lead in these matrixes by FPIA. The limit of detection under conditions that minimized sample dilution was approximately 1 ppb, and cross reactivity with 15 nontarget metals was below 0.5% in all cases. The methods are simple to perform and are amenable to field testing and mobile laboratory use, allowing timely and cost effective characterization of suspected sources of lead contamination. PMID- 11917990 TI - Automated analysis of nanomolar concentrations of phosphate in natural waters with liquid waveguide. AB - Concentrations of phosphate in natural waters are often below the detection limits of conventional nutrient autoanalyzers, by either gas-segmented continuous flow analysis orflow injection analysis. A liquid waveguide capillary flow cell has been used to extend the sensitivity of a conventional autoanalyzer for the automated analysis of nanomolar concentrations of phosphate in natural waters. Total reflection of light can be achieved within the liquid core of the flow cell because the refractive index of a cell wall coated with Teflon 1600 is lower than that of water. This property allows the manufacturers to construct long liquid waveguide capillary flow cells in a helical, rather than a linear shape, with compact dimensions. A small sample volume is required because the internal volume of a 2-m long capillary flow cell is only approximately 0.5 cm3. Adaptation of this long flow cell to autoanalyzers significantly enhances the sensitivity of automated colorimetric analysis of phosphate with a molybdenum blue method, allowing for the accurate and precise determination of nanomolar concentrations of phosphate in natural waters. The advantages of this technique are a low detection limit (0.5 nM), a small sample volume (2 mL), high precision (2% at 10 nM levels), and automation for the rapid analysis of a large number of samples. PMID- 11917991 TI - Immobilizing humic acid in a sol-gel matrix: a new tool to study humic contaminants sorption interactions. AB - Humic substances originated from aquatic, soil, or sediment environments are mixtures of humic compounds with various characteristics. Sorption interactions with isolated, well defined humic fractions can be studied either in an aqueous phase ("dissolved humic substances"), or in a solid-phase, by coating mineral particles with the humic materials, or simply by working with humic acid particles (powder) at low pH to minimize dissolution. Each attitude, by definition, can be studied by different experimental techniques and has a different meaning for understanding natural environmental processes. In this study, a new tool for studying sorption interactions is presented. Sol-gel was used as an inert matrix to immobilize (entrap) various humic acids (HAs), and then used to study the interactions of several polycyclic aromatic hydrocarbons (PAHs) with the entrapped HA. Linear and nonlinear sorption coefficients were highly correlated with contaminant hydrophobicity. Sorption of pyrene to immobilized HA was in the order of soil HA > Aldrich HA approximately = peat HA. It was concluded that the entrapped HAs retained their original properties in the gel matrix and were accessible to the external contaminant through the pore network. Additionally, binding coefficients of pyreneto dissolved humic substances and to dissolved organic matter (DOM) were determined from the reduction in pyrene sorption to immobilized HA in the presence of dissolved humic material or DOM in solution. Binding coefficients of pyrene were in the order of the following: dissolved Aldrich HA > dissolved peat fulvic acid (FA) > DOM derived from mature compost > DOM derived from fresh compost. PMID- 11917992 TI - Comparison of copper speciation in coastal marine waters measured using analytical voltammetry and diffusion gradient in thin-film techniques. AB - The diffusion gradient in thin-film hydrogel (DGT) probe is a promising tool for metal speciation work. Based on a passive sampling principle, it provides the potential for large data sets in complex regimes. DGT probes were deployed in waters characterized independently using competitive ligand exchange-adsorptive cathodic stripping voltammetry (CLE-ACSV). The CLE-ACSV used benzoyl acetone as the competitive ligand in discrete water samples collected during the deployment of the DGT probes. The DGT probes used a 15% polyacrylamide/0.4% bis-acrylamide cross-linker hydrogel and a Na-form of Chelex-100 to complex metal that fluxed into the probe through the hydrogel. Probes were deployed in locations characterized by the degree of pollution impact: the relatively pristine Vineyard Sound, MA, [Cu]total approximately 6 nM, small seasonally active harbors on Cape Cod, MA, [Cu]total = 12-64 nM, as well as a large polluted estuary, the Elizabeth River, VA, [Cu]total = 44-58 nM, and a large polluted port, San Diego Harbor, CA, [Cu]total = 23-103 nM. This is the first study where DGT probes have been compared with an independent speciation technique in marine systems and used to establish the diffusion coefficient of Cu-complexing ligands in situ. Results showed that the probes produced highly precise data sets, with substantial differences in copper accumulation between contaminated and pristine waters. Comparison of DGT results with CLE-CSV indicate that at least 10-35% of the organically complexed copper derived by CLE-ACSV measurements was DGT-labile. Diffusion coefficients (corrected to 25 degrees C) of organically complexed DGT labile Cu through the hydrogel ranged from 0.77 x 10(-6) cm2 s(-1) in Vineyard Sound to 2.16 x 10(-6) cm2 s(-1) in the Elizabeth River estuary. Accumulation rates of copper were substantially higher in contaminated waters than in pristine waters, suggesting that the probes in their current form may be useful as tracking tools to detect episodic sources of contamination. PMID- 11917993 TI - Optimization of method for detecting and characterizing NOM by HPLC-size exclusion chromatography with UV and on-line DOC detection. AB - High-performance liquid chromatography (HPLC)-size exclusion chromatography (SEC) with ultraviolet absorbance (UVA) and on-line dissolved organic carbon (DOC) detectors has been adapted and optimized under various conditions. An enhanced HPSEC-UVA system employing a modified commercially available DOC detector provides a better understanding of the qualitative and quantitative natural organic matter (NOM) properties in water samples by detecting aromatic and nonaromatic fractions of NOM as a function of molecular weight (MW). The most critical merit of this system is that the DOC detector is readily available and widely used. With only a few modifications, a commercially available TOC analyzer served as a DOC detector, integrated with the HPSEC to measure DOC along with UVA, and provided a specific UVA (SUVA) chromatograph that is useful information for drinking water plant design and operation. Without preconcentration, samples can be analyzed with a small amount of sample, with a DOC detection limit as low as 0.1 mg/L (as DOC). PMID- 11917994 TI - A case study to detect the leakage of underground pressureless cement sewage water pipe using GPR, electrical, and chemical data. AB - The exploration and determination of leakage of underground pressureless nonmetallic pipes is difficult to deal with. A comprehensive method combining Ground Penetrating Rader (GPR), electric potential survey and geochemical survey is introduced in the leakage detection of an underground pressureless nonmetallic sewage pipe in this paper. Theoretically, in the influencing scope of a leakage spot, the obvious changes of the electromagnetic properties and the physical chemical properties of the underground media will be reflected as anomalies in GPR and electrical survey plots. The advantages of GPR and electrical survey are fast and accurate in detection of anomaly scope. In-situ analysis of the geophysical surveys can guide the geochemical survey. Then water and soil sampling and analyzing can be the evidence for judging the anomaly is caused by pipe leakage or not. On the basis of previous tests and practical surveys, the GPR waveforms, electric potential curves, contour maps, and chemical survey results are all classified into three types according to the extent or indexes of anomalies in orderto find out the leakage spots. When three survey methods all show their anomalies as type I in an anomalous spot, this spot is suspected as the most possible leakage location. Otherwise, it will be down grade suspected point. The suspect leakage spots should be confirmed by referring the site conditions because some anomalies are caused other factors. The excavation afterward proved that the method for determining the suspected location by anomaly type is effective and economic. Comprehensive method of GRP, electric potential survey, and geochemical survey is one of the effective methods in the leakage detection of underground nonmetallic pressureless pipe with its advantages of being fast and accurate. PMID- 11917995 TI - Adsorption characteristics of SO2 on activated carbon prepared from coconut shell with potassium hydroxide activation. AB - The adsorption characteristics of SO2 were studied with KOH-impregnated granular activated carbon (K-IAC). To confirm selective SO2 adsorptivity of K-IAC using a fixed bed adsorption column, experiments were conducted on the effects of KOH and of linear velocity, temperature, and concentration. In addition, changes in features before and after adsorption were observed by utilizing FTIR, XRD, ToF SIMS, and AES/SAM, examining the surface chemistry. K-IAC adsorbed 13.2 times more SO2 than did general activated carbon (GAC). The amount of SO2 adsorbed increased as linear velocity and concentration increased and as temperature decreased. At lower temperature, the dominant reaction between KOH and SO2 produces K2-SO3 and H2O. Any H2O remaining on the surface is converted into H2SO4 as SO2 and O2 are introduced. Then, the KOH and SO2 reaction produces K2SO4 and H2O. The surface characterization results proved that adsorption occurred through chemical reaction between KOH and SO2. The SO2 adsorbed K-IAC exists in the form of stable oxide crystal, K2SO3 and K2SO4, due to potassium. The basic feature given to the surface of activated carbon by KOH impregnation was confirmed to be acting as the main factor in enhancing SO2 adsorptivity. PMID- 11917996 TI - Evaluating remedial alternatives for an acid mine drainage stream: application of a reactive transport model. AB - A reactive transport model based on one-dimensional transport and equilibrium chemistry is applied to synoptic data from an acid mine drainage stream. Model inputs include streamflow estimates based on tracer dilution, inflow chemistry based on synoptic sampling, and equilibrium constants describing acid/base, complexation, precipitation/dissolution, and sorption reactions. The dominant features of observed spatial profiles in pH and metal concentration are reproduced along the 3.5-km study reach by simulating the precipitation of Fe(III) and Al solid phases and the sorption of Cu, As, and Pb onto freshly precipitated iron(III) oxides. Given this quantitative description of existing conditions, additional simulations are conducted to estimate the streamwater quality that could result from two hypothetical remediation plans. Both remediation plans involve the addition of CaCO3 to raise the pH of a small, acidic inflow from approximately 2.4 to approximately 7.0. This pH increase results in a reduced metal load that is routed downstream by the reactive transport model, thereby providing an estimate of post-remediation water quality. The first remediation plan assumes a closed system wherein inflow Fe(II) is not oxidized by the treatment system; under the second remediation plan, an open system is assumed, and Fe(II) is oxidized within the treatment system. Both plans increase instream pH and substantially reduce total and dissolved concentrations of Al, As, Cu, and Fe(II+III) at the terminus of the study reach. Dissolved Pb concentrations are reduced by approximately 18% under the first remediation plan due to sorption onto iron(III) oxides within the treatment system and stream channel. In contrast, iron(III) oxides are limiting under the second remediation plan, and removal of dissolved Pb occurs primarily within the treatment system. This limitation results in an increase in dissolved Pb concentrations over existing conditions as additional downstream sources of Pb are not attenuated by sorption. PMID- 11917997 TI - Removal of sulfur-organic polar micropollutants in a membrane bioreactor treating industrial wastewater. AB - While membrane bioreactors (MBR) have proven their large potential to remove bulk organic matter from municipal as well as industrial wastewater, their suitability to remove poorly degradable polar wastewater contaminants is yet unknown. However, this is an important aspect for the achievable effluent quality and in terms of wastewater reuse. We have analyzed two classes of polar sulfur-organic compounds, naphthalene sulfonates and benzothiazoles, by liquid chromatography electrospray ionization-tandem mass spectrometry (LC-ESI-MS) over a period of 3 weeks in the influent and effluent of a full-scale MBR with external ultrafiltration that treats tannery wastewater. While naphthalene monosulfonates were completely removed, total naphthalene disulfonate removal was limited to about 40%, and total benzothiazoles concentration decreased for 87%. Quantitative as well as qualitative data did not indicate an adaptation to or a more complete removal of these polar aromatic compounds by the MBR as compared to literature data on conventional activated sludge treatment. While quality improvements in receiving waters for bulk organic matter are documented and the same can be anticipated for apolar particle-associated contaminants, these data provide no indication that MBR will improve the removal of polar poorly biodegradable organic pollutants. PMID- 11917998 TI - Optimal design of zero-water discharge rinsing systems. AB - This paper is about zero liquid discharge in processes that use water for rinsing. Emphasis was given to those systems that contaminate process water with valuable process liquor and compounds. The approach involved the synthesis of optimal rinsing and recycling networks (RRN) that had a priori excluded water discharge. The total annualized costs of the RRN were minimized by the use of a mixed-integer nonlinear program (MINLP). This MINLP was based on a hyperstructure of the RRN and contained eight counterflow rinsing stages and three regenerator units: electrodialysis, reverse osmosis, and ion exchange columns. A "large-scale nickel plating process" case study showed that by means of zero-water discharge and optimized rinsing the total waste could be reduced by 90.4% at a revenue of $448,000/yr. Furthermore, with the optimized RRN, the rinsing performance can be improved significantly at a low-cost increase. In all the cases, the amount of valuable compounds reclaimed was above 99%. PMID- 11917999 TI - Mathematical model for meso- and thermophilic anaerobic sewage sludge digestion. AB - A mathematical model is developed to describe the dynamic behavior of mesophilic (35 +/- 5 degrees C) and thermophilic digestion (55 +/- 5 degrees C). Special emphasis is given to acetotrophic methanogenesis and propionate degradation, as the steps that determine the stability of anaerobic digestion, as well as to hydrolysis rate, which determines the degradation efficiency of particulate degradable organic carbon. Within the range of 6-20 (mesophilic) and 2-8 d (thermophilic) hydraulic retention time (HRT), the observed maximum growth rates for acetotrophic methanogens are 0.33 and 1.3 d(-1), respectively, with a 15% decay rate. Temperature and pH dependence as well as ammonia inhibition of acetate and propionate conversion are determined and included in the model, which allows us to simulate the effect of protein- and nitrogen-rich waste addition and the consequences of temporarily increased free ammonia at high pH. No inhibition of hydrogen conversion was observed in the same free ammonia range. The pH optimum is between 6.6 and 7.3. Acetotrophic methanogenesis is strongly inhibited below pH 6.2, whereas above pH 7.4 it can be inhibited by free ammonia. For digesters fed with ordinary municipal sewage sludge, free ammonia inhibition of acetate conversion leads to an increase in acetate at about 35 and 140 mg of N/L for mesophilic (HRT = 20 d) and thermophilic (HRT = 6 d) conditions, respectively. The hydrolysis rate constant is 0.25 and 0.4 d(-1) respectively for these two conditions. The model is validated with load variation experiments in laboratory and full-scale digesters for step and shock loads. PMID- 11918000 TI - Products of pertechnetate radiolysis in highly alkaline solution: structure of TcO2 x xH2O. AB - The chemistry of technetium in certain high-level nuclear waste (HLW) tanks at the Hanford Site complicates the treatment and vitrification of HLW. A major problem is the presence, in certain tanks, of unidentified, lower-valent technetium species, which are difficult to remove from the waste by current separation processes. Radiolytic reduction of TcO4- in alkaline solutions containing selected organic compounds, approximating the conditions in HLW, was investigated to determine the classes of compounds that can be formed under these conditions. Insoluble TcO2 x xH2O is the primary radiolysis product with the majority of organic compounds investigated, including citrate, dibutyl phosphate, and aminopolycarboxylates. X-ray absorption fine structure (XAFS) measurements show that TcO2 x xH2O has a one-dimensional chain structure consisting of edge sharing TcO6 octahedra with bridging oxide and trans water ligands. When diols, such as ethylene glycol, are present, only soluble, Tc(IV) alkoxide compounds are produced. The XAFS and UV-visible spectra of these compounds provide evidence for a binuclear structure similar to (H2EDTA)2Tc2(mu-O)2. The properties of the Tc(IV) alkoxide complexes were determined and are consistent with those observed for the soluble, lower-valent technetium complexes that complicate the treatment of HLW at the Hanford site. PMID- 11918001 TI - Quantitative assessment of solid waste treatment systems in the industrial ecology perspective by exergy analysis. AB - Solid waste treatment options (recycling, incineration, and landfilling; the two latter processes both with co-generation of heat and electricity) have been studied for cardboard, newspaper, polyethylene, poly(ethylene terephthalate), polypropylene, polystyrene, and poly(vinyl chloride) waste. The conversion processes have been analyzed in terms of the second law of thermodynamics. The analysis allows calculating the exergy (useful energy) embodied in conversion products that can be obtained from the required inputs for the treatment processes. Taking into account the waste materials and the resources to convert them, it proved that recycling is the most efficient option for polyethylene with an efficiency of 62.5% versus 43.6% for incineration and 0.9% for landfilling. Next, waste treatment has been put into the broader perspective of industrial ecology. Exergetic efficiencies of industrial metabolic options have been calculated. Here resources for manufacturing and converting solid products have been considered. Furthermore, selection of one type of conversion excludes the generation of other potential conversion products. Therefore, it has to be taken into account that these latter products still have to be produced starting from virgin resources. Recycling proved to be the most efficient strategy: the ratio eta between exergy embodied in all delivered products on one hand, and all exergy withdrawn from the ecosphere or from waste materials on the other hand, is the highest. For polyethylene, eta proved to be 0.568, whereas eta is 0.503 and 0.329 for incineration and landfilling, respectively. On the other hand, if R the ratio between exergy of delivered products on one hand and exergy of virgin materials on the other hand is calculated, the differences between the industrial metabolic options are larger. Recycling polyethylene showed a ratio R of 0.936, whereas ratios of 0.772 and 0.531 were found for incineration and landfilling, respectively. It has been shown that the exergy concept allows a quantitative comparison of different industrial metabolic options, contributing to a better assessment of sustainability of technology with respect to resource management. PMID- 11918002 TI - Adsorption and pressure swing desorption of NOx in Na-Y zeolite: experiments and modeling. AB - Pressure swing NOx adsorption-desorption cycles were performed in the temperature range 200-350 degrees C using a fixed adsorbent bed of compressed Na-Y pellets and using a honeycomb coated with Na-Y powder. The experiments were performed using a synthetic gas mixture mimicking exhaust from a lean burn internal combustion engine. Na-Y zeolite coadsorbs NO and NO2 as N2O3, which in the regeneration were displaced by competitively adsorbed water molecules from a hydrated air stream. The performance of the fixed bed in these NOx adsorption and displacement desorption processes were modeled with a one-dimensional model. The kinetic and thermodynamic parameters from the fixed bed model were implemented in a model for the operation of the monolith. The experimental adsorption and desorption NOx concentration profiles in the monolith were reasonably well reproduced by the model. The water content of the flushing stream and the stripping gas flow rate are key process parameters. Technically, both parameters can be optimized in a valveless system with rotating honeycomb adsorbent comprising a NOx adsorption, a water injection and a NOx evacuation section. PMID- 11918003 TI - Freshwater bivalve shells as archival indicators of metal pollution from a copper uranium mine in tropical northern Australia. AB - Freshwater bivalves (Velesunio angasi) were sampled in 1996 from the Finniss River in tropical northern Australia at 10 sites a priori exposed and nonexposed to acid rock drainage (ARD), containing elevated metal concentrations, from the rehabilitated Rum Jungle copper-uranium mine. Secondary ion mass spectrometry (SIMS) was used to measure Cu, Mn, Zn, U, Ni, Co, Pb, and Fe/Ca ratios across the annual shell laminations of the longest-lived bivalves found at each site, with the aim of evaluating the ability of the shells to archive measured annual metal inputs and their temporal patterns. At sites not contaminated by ARD, relatively constant and similar (baseline) SIMS signals were found for all metals in the shell laminations of V. angasi, dating as far back as 1965. At sites contaminated by ARD, relatively constant, but variably elevated, SIMS signals were evident for Cu, Mn, Zn, Ni, and Co in the shell, which extended back to the end of rehabilitation (1986) only. Since rehabilitation, the temporal patterns of Cu, Zn, and Mn observed in the shells at the most contaminated sites reflected those of the measured annual dissolved loads in the surface waters. The average concentrations of Cu, Mn, Zn, Ni, and Co in the shells decreased (3-13-fold) with increasing distance downstream of the mine site, until concentrations characteristic of the noncontaminated sites were reached. This geographic pattern of decline in pollution signal in the shell with increasing distance downstream of the pollution input is consistent with the pattern established for water and sediment chemistry. Overall, the SIMS results support the proposition that the shells of V. angasi can be used as archival indicators of metal pollution in surface waters of the Finniss River over their lifetime. PMID- 11918004 TI - Emission of polycyclic aromatic hydrocarbons, toxicity, and mutagenicity from domestic cooking using sawdust briquettes, wood, and kerosene. AB - Smoke samples, in both gas and particulate matter (PM) phases, of the three domestic stoves were collected using U.S. EPA modified method 5 and were analyzed for 17 PAH (HPLC-UV), acute toxicity (Microtox test), and mutagenicity (Amestest). The gas phase of smoke contributed > or = 95% of 17 PAH, > or = 96% of toxicity, and > or = 60% of mutagenicity. The highest emission factor of 17 PAH was from sawdust briquettes (260 mg/kg), but the highest emission of 11 genotoxic PAH was from kerosene (28 mg/kg). PM samples of kerosene smoke were not toxic. The total toxicity emission factor was the highest from sawdust, followed by kerosene and wood fuel. Smoke samples from the kerosene stove were not mutagenic. TA98 indicated the presence of both direct and indirect mutagenic activities in PM samples of sawdust and wood fuel but only direct mutagenic activities in the gas phase. TA100 detected only direct mutagenic activities in both PM and gas-phase samples. The higher mutagenicity emission factor was from wood fuel, 12 x 10(6) revertants/kg (TA100-S9) and 3.5 x 10(6) (TA98-S9), and lower from sawdust, 2.9 x 10(6) (TA100-S9) and 2.8 x 10(6) (TA98-S9). The low burning rate and high efficiency of a kerosene stove have resulted in the lowest PAH, toxicity, and mutagenicity emissions from daily cooking activities. The bioassays produced toxicity and mutagenicity results in correspondence with the PAH content of samples. The tests could be used for a quick assessment of potential health risks. PMID- 11918005 TI - Photochemical changes in cyanide speciation in drainage from a precious metal ore heap. AB - In drainage from an inactive ore heap at a former gold mine, the speciation of cyanide and the concentrations of several metals were found to follow diurnal cycles. Concentrations of the hexacyanoferrate complex, iron, manganese, and ammonium were higher at night than during the day, whereas weak-acid-dissociable cyanide, silver, gold, copper, nitrite, and pH displayed the reverse behavior. The changes in cyanide speciation, iron, and trace metals can be explained by photodissociation of iron and cobalt cyanocomplexes as the solutions emerged from the heap into sunlight-exposed channels. At midday, environmentally significant concentrations of free cyanide were produced in a matter of minutes, causing trace copper, silver, and gold to be mobilized as cyanocomplexes from solids. Whether rapid photodissociation is a general phenomenon common to other sites will be important to determine in reaching a general understanding of the environmental risks posed by routine or accidental water discharges from precious metal mining facilities. PMID- 11918006 TI - Gas-phase organics in environmental tobacco smoke. 1. Effects of smoking rate, ventilation, and furnishing level on emission factors. AB - We measured the emissions of 26 gas-phase organic compounds in environmental tobacco smoke (ETS) using a model room that simulates realistic conditions in residences and offices. Exposure-relevant emission factors (EREFs), which include the effects of sorption and re-emission over a 24-h period, were calculated by mass balance from measured compound concentrations and chamber ventilation rates in a 50-m3 room constructed and furnished with typical materials. Experiments were conducted at three smoking rates (5, 10, and 20 cigarettes day(-1)), three ventilation rates (0.3, 0.6, and 2 h(-1)), and three furnishing levels (wallboard with aluminum flooring, wallboard with carpet, and full furnishings). Smoking rate did not affect EREFs, suggesting that sorption was linearly related to gas phase concentration. Furnishing level and ventilation rate in the model room had little effect on EREFs of several ETS compounds including 1,3-butadiene, acrolein, acrylonitrile, benzene, toluene, and styrene. However, sorptive losses at low ventilation with full furnishings reduced EREFs for the ETS tracers nicotine and 3-ethenylpyridine by as much as 90 and 65% as compared to high ventilation, wallboard/aluminum experiments. Likewise, sorptive losses were 40 70% for phenol, cresols, naphthalene, and methylnaphthalenes. Sorption persisted for many compounds; for example, almost all of the sorbed nicotine and most of the sorbed cresol remained sorbed 3 days after smoking. EREFs can be used in models and with ETS tracer-based methods to refine and improve estimates of exposures to ETS constituents. PMID- 11918007 TI - Records of change in salt marshes: a radiochronological study of three Westerschelde (SW Netherlands) marshes. AB - Three salt marshes on a 50-km transect along the north bank of the Westerschelde Estuary were investigated to determine whether salt marshes in the estuary had responded to shipping channel modifications in recent decades. Marsh accretion rates were estimated mainly from 137Cs profiles with further evidence from 241Am because changes in both rate of deposition and nature of the accreting material precluded use of standard 210Pb(excess) dating models. The 137Cs profiles usually show peaks corresponding to atmospheric deposition from the 1963 fallout maximum and sometimes from the Chernobyl accident, although intervening enhanced 137Cs activities derived from the nuclear reprocessing marine discharges of Sellafield and La Hague are clearly discernible. In all three marshes (Ritthem at the mouth of the estuary and Zuidgors and Waarde at 20 and 45 km upstream), a marked, near coincident change in the rate of accumulation and in the grain size of material deposited occurred around 1980. This may be related to a combination of channel deepening and straightening operations undertaken in the mid-1970s and/or natural changes in winter wave climate. PMID- 11918008 TI - Temporal trends of organochlorine pesticides in the Canadian Arctic atmosphere. AB - Temperature normalization (TN), multiple linear regression (MLR), and digital filtration (DF) were used to analyze the temporal trends of an atmospheric dataset on organochlorine pesticides (OCs) collected at the Canadian high arctic site of Alert, Nunavut. Details of these techniques have been presented before (Environ. Sci. Technol. 2001, 35, 1303-1311). Both the TN and DF methods revealed that the majority of OC pesticides declined over the 5 years of study, except endosulfan I and several of the pesticide metabolites, including dieldrin and p,p'-DDE. In comparison to studies conducted in the Great Lakes, atmospheric levels in the Arctic were less dependent on temperature, although seasonal variations were apparent. Generally, levels in the winter were lower than during the rest of the year. A notable exception was p,p'-DDE. Many compounds also showed a second minimum in concentrations during June/July and possible explanations are presented to account for this. The estimated first order half lives for the decline in OC concentrations were generally found to be comparable or slightly longer than those obtained at temperate locations, with the exception of alpha-HCH, which displayed a much longer half-life in the Arctic (approximately 17 yrs). Sporadic increases in heptachlor as well as increases in the ratio of trans- to cis-chlordane suggest episodic input of chlordanes between 1995 and 1997, especially during the winter. PMID- 11918009 TI - Ozonation of chrysene: evaluation of byproduct mixtures and identification of toxic constituent. AB - The effects of chrysene and the ozonated byproducts on in vitro gap junctional intercellular communication (GJIC) were evaluated using the scrape loading/dye transfer (SL/DT) technique. A 1 mM solution of chrysene was ozonated at dosages of 1.75, 3, 4.25, and 5 mol O3/mol chrysene (Chr). The early ozonation mixture, 1.75 mol O3/mol Chr, exhibited greater inhibition to GJIC than chrysene and irreversible damage to cells leading to cell death. To determine the compounds potentially responsible for the increase in toxicity, the byproducts formed upon treatment with 1.44 mol O3/mol Chr were separated into 14 fractions using RP HPLC. The major compounds identified in the fractions were 2-(2'-formyl) phenyl-1 naphthaldehyde, 2-(2'formyl) phenyl-1-naphthoic acid, and 2-2-carboxyphenyl-1 naphthoic acid. 2-(2'-Formyl) phenyl-1-naphthaldehyde was determined to be the compound causing GJIC inhibition in sample fractions and byproduct mixtures. PMID- 11918010 TI - Effects of dietary methylmercury on reproduction of fathead minnows. AB - We examined effects of dietary methylmercury (MeHg) on reproduction of fathead minnows (Pimephales promelas). Juvenile fish were fed one of four diets until sexual maturity (phase 1): a control diet (0.06 microg Hg g(-1) dry weight) and three diets contaminated with MeHg at 0.88 (low), 4.11 (medium), and 8.46 microg Hg g(-1) dry weight (high). At sexual maturity, male and female fish were paired, again fed one of the four diets, and allowed to reproduce (phase 2). To assess effects of MeHg during gametogenesis, some fish were fed diets during phase 2 that differed from those during phase 1. Spawning success of pairs fed the same diet during phases 1 and 2 was 75% for controls and 46%, 50%, and 36% for the low , medium-, and high-MeHg treatments, respectively. Spawning success of pairs fed a contaminated diet during phase 1 and a control diet during phase 2 was 63%, 40%, and 14% for the low-, medium-, and high-MeHg treatments, respectively, whereas exposure to dietary MeHg only during phase 2 did not reduce spawning success. Dietary MeHg delayed spawning, and days to spawning was positively correlated with concentration of total mercury in the carcasses of test fish. MeHg reduced the instantaneous rate of reproduction of fish fed the same diets during phases 1 and 2. Both the gonadosomatic index and reproductive effort of female fish were inversely correlated with mercury in carcasses, whereas developmental and hatching success of embryos, 7-d survival, and 7-d growth of larvae were unrelated to mercury concentrations in parental fish or their diets. MeHg decreased reproduction of adult fathead minnows at dietary concentrations encountered by predatory fishes in aquatic systems with MeHg-contaminated food webs, implying that exposed fish populations could be adversely affected by this widespread contaminant. PMID- 11918011 TI - Chlorination of phenols: kinetics and formation of chloroform. AB - The kinetics of chlorination of several phenolic compounds and the corresponding formation of chloroform were investigated at room temperature. For the chlorination of phenolic compounds, second-order kinetics was observed, first order in chlorine, and first-order in the phenolic compound. The rate constants of the reactions of HOCl with phenol and phenolate anion and the rate constant of the acid-catalyzed reaction were determined in the pH range 1-11. The second order rate constants for the reaction HOCl + phenol varied between 0.02 and 0.52 M(-1) s(-1), for the reaction HOCl and phenolate between 8.46 x 10(1) and 2.71 x 10(4) M(-1) s(-1). The rate constant for the acid-catalyzed reaction varied between 0.37 M(-2) s(-1) to 6.4 x 10(3) M(-2) s(-1). Hammett-type correlations were obtained for the reaction of HOCl with phenolate (log(k) = 4.15-3.00 x sigma sigma) and the acid-catalyzed reaction of HOCl with phenol (log(k) = 2.37-4.26 x sigma sigma). The formation of chloroform could be interpreted with a second order model, first-order in chlorine, and first-order in chloroform precursors. The corresponding rate constants varied between k > 100 M(-1) s(-1) for resorcinol to 0.026 M(-1) s(-1) for p-nitrophenol at pH 8.0. It was found that the rate-limiting step of chloroform formation is the chlorination of the chlorinated ketones. Yields of chloroform formation depend on the type and position of the substituents and varied between 2 and 95% based on the concentration of the phenol. PMID- 11918012 TI - Relative dominance of physical versus chemical effects on the transport of adhesion-deficient bacteria in intact cores from South Oyster, Virginia. AB - Bacterial transport experiments were conducted using intact sediment cores collected from sites on the Delmarva Peninsula near South Oyster, VA, to delineate the relative importance of physical and chemical heterogeneity in controlling transport of an adhesion-deficient bacterial strain. Electron microscopy revealed that the sediments consisted of quartz and feldspar with a variable amount of clay and iron and aluminum hydroxide coatings on the grains. A nonmotile, gram-negative indigenous groundwater strain, designated as Comamonas sp. DA001, was injected into the cores along with a conservative tracer bromide (Br). DA001 cells were 1.2 x 0.6 microm in size with a hydrophilic surface and a slightly negative surface charge. Bacterial breakthrough preceded that of Br. This differential advection phenomenon can be accounted for by reduction of the effective porosity for the bacteria relative to Br. The distribution of cells remaining in the core as determined by scintillation counting and phosphor imaging techniques was variable, ranging from nearly uniform concentrations throughout the core to exponentially decreasing concentrations with distance from the point of injection. The fraction of bacterial retention in the core was positively correlated with the abundance of the metal hydroxides and negatively correlated with grain size. Because grain size was inversely correlated with the abundance of the metal hydroxide coatings, it was necessary to separate the effects of grain size and mineralogy. The fraction of the bacterial retention accounting for the effect of grain size, the collision efficiency, exhibited no correlation with the abundance of the metal hydroxides, indicating that the bacterial retention was primarily controlled by grain size. Reasons for the lack of influence of mineralogy on bacterial transport include (i) the slightly negatively charged bacterial surfaces; (ii) an insufficient heterogeneity of sediment surface properties; and (iii) the masking of the positive charge of the metal hydroxide surfaces by adsorbed organic carbon (up to 1180 ppm). This study demonstrates that the laboratory-based bacterial transport experiments are effective in delineating physical versus chemical controlling factors and provide an important link to field-based bacterial transport studies. PMID- 11918013 TI - PBDE information overlooked? PMID- 11918014 TI - Industry should assist TMDL development. PMID- 11918015 TI - Reduction of hexavalent chromium by H2O2 in acidic solutions. AB - The rates of the reduction of Cr(VI) with H2O2 were measured in NaCl solutions as a function of pH (1.5-4.8), temperature (5-40 degrees C), and ionic strength (I = 0.01-2 M) in the presence of an excess of reductant. The rate of Cr(VI) reduction is described by the general expression -d[Cr(VI)]/dt = k2[Cr(VI)](m)[H2O2](n)[H+](z), where m = 1 and n and z are two interdependent variables. The value of n is a function of pH between 2 and 4 (n = (3 x 10(a))/(1 + 10(a)), where a = -0.25 - 0.58pH + 0.26pH2) leveling off at pH < 2 (where n approximately = 1) and pH > 4 (where n approximately = 3). The rates of Cr(VI) reduction are acid-catalyzed, and the kinetic order z varies from about 1.8-0.5 with increasing H2O2 concentration, according to the equation z = 1.85 - 350.1H2O2 (M) which is valid for [H2O2] < 0.004 M. The values of k2 (M(-(n+z)) min(-1)) are given by k2 = k/[H+](z) = k1/[H2O2](n)[H+](z), where k is the overall rate constant (M(-n) min(-1)) and k, is the pseudo-first-order rate constant (min(-1)). The values of k in the pH range 2-4 have been fitted to the equation log k = 2.14pH - 2.81 with sigma = +/- 0.18. The values of k2 are dependent on pH as well. Most of the results with H2O2 < 3 mM are described by log k2 = 2.87pH - 0.55 with sigma = +/- 0.54. Experimental results suggest that the reduction of Cr(VI) to Cr(III) is controlled by the formation of Cr(V) intermediates. Values of k2 and k calculated from the above equations can be used to evaluate the rates of the reaction in acidic solutions under a wide range of experimental conditions, because the rates are independent of ionic strength, temperature, major ions, and micromolar levels of trace metals (Cu2+, Ni2+, Pb2+). The application of this rate law to environmental conditions suggests that this reaction may have a role in acidic solutions (aerosols and fog droplets) in the presence of high micromolar concentrations of H2O2. PMID- 11918016 TI - Poorly functioning sewage treatment plants? PMID- 11918017 TI - Species kinetics and heterogeneous reactivity of dissolved Cu in natural freshwaters. AB - Using high specific activity 64Cu2+ as radiotracer, the distribution kinetics among Cu species were established in natural organic-rich freshwaters under steady-state conditions, i.e., with minimal disturbance of existing equilibria. Study sites with contrasting suspended particulate matter (SPM) characteristics were investigated. Our analytical protocol allowed the differentiation between the following Cu species: SPM associated Cu, dissolved reactive (free and labile) Cu, and organically complexed Cu. The data obtained were successfully evaluated by compartmental analysis, which showed the importance of organically complexed Cu in freshwaters, and the dominant role of the interactions between organically complexed Cu and SPM in a SPM-rich water. The kinetic 54Cu measurements indicated that the attainment of equilibrium between dissolved reactive and organically complexed Cu took ca. <1-2 h, and 4-15 h for the interaction between dissolved organically complexed and SPM associated Cu. The kinetic study was augmented by voltammetric measurements of the dissolved (stable) Cu equilibrium speciation conditions in the natural waters. These measurements showed that the waters contained very low cupric ion concentrations (10(-12)-10(-15) M), with more than 99.9% of the dissolved Cu complexed by strong organic ligands (conditional stability constants: 10(13.4)-10(15.4)). PMID- 11918018 TI - Recovery of streams from episodic acidification in northern Sweden. AB - Between 1990 and 1999, SO4(2-) deposition in northern Sweden decreased by over 50%. To determine if a corresponding amelioration of stream acidity has occurred, we analyzed trends in anthropogenically driven episodic acidification in five streams during the same time period, using the Boreal Dilution Model (BDM) (Bishop, K. H.; Laudon, H.; Kohler, S. Water Resour. Res. 2000, 36, 1873-1884). Although there was no significant change in the annual average streamwater chemistry, the anthropogenically driven episodic acidification associated with spring flood runoff decreased by between 40% and 80%. A strong correlation between winter SO4(2-) deposition and the anthropogenic component of episodic acidification in these five streams suggests that future reductions of acid deposition will further improve the spring flood acidification situation in northern Sweden. These results argue that reduced emissions of acid precursors have generated significant improvements in the surface water chemistry during episodes associated with spring runoff in northern Sweden. PMID- 11918019 TI - Correlation of poly(methylene)-rich amorphous aliphatic domains in humic substances with sorption of a nonpolar organic contaminant phenanthrene. AB - The structural makeup of natural organic matter plays a major role in regulating its capacity to retain nonionic hydrophobic organic compounds (HOCs). We used a model HOC--phenanthrene--to investigate the correlations between sorption capacity, specifically the modified Freundlich coefficient (K'f), and compositional data of humic acids, humins, and a peat obtained from quantitative 13C solid-state NMR spectroscopy. A positive correlation between K'f and the weight fraction of amorphous poly(methylene) in the sorbents was observed. In contrast, the correlation between phenanthrene sorption capacity and aromaticity or polarity indices of the sorbents was insignificant. The nonpolar aliphatic carbon fraction, excluding poly(methylene), was only partially correlated with K'f. Detailed NMR analyses of the sorbents using 1H inversion-recovery experiments showed that 10-nm diameter domains of branched nonpolar aliphatic groups, which account for 20-50% of all nonpolar aliphatic segments and may be associated with the poly(methylene), were responsible for the partial correlation. The correlation between K'f and the amorphous nonpolar aliphatic domains including amorphous poly(methylene) was strong. The rubbery, relatively low-density, and amorphous nonpolar aliphatic domains can be expected to offer an excellent environment for the sorption of phenanthrene by partitioning. These observations suggest that the domains of amorphous poly(methylene) and branched nonpolar aliphatics, which make up 2-9 wt % of the organic fraction in our samples, may serve as good descriptors for the potential of natural organic matter to retain HOCs in the natural environments. PMID- 11918020 TI - U(VI) adsorption to heterogeneous subsurface media: application of a surface complexation model. AB - The pH-dependent adsorption of U(VI) onto three heterogeneous, subsurface media from the Department of Energy (DOE) Oak Ridge Reservation, Savannah River Site, and Hanford Reservation was investigated. The three materials contained significant quantities of iron and manganese oxides with nearly identical extractable iron oxide contents (25.3-25.8 g/kg). A model independently developed for the adsorption of U(VI) to synthetic ferrihydrite (Waite, T. D.; Davis, J. A.; Payne, T. E.; Waychunas, G. A.; Xu, N. Geochim. Cosmochim. Acta 1994, 58, 5465-5478) was able to predict the major features of the pH-dependent U(VI) adsorption to the materials under the assumption that all the dithionite-citrate bicarbonate extractable iron oxide was present as ferrihydrite. Further experiments with the Oak Ridge soil as a function of carbonate and U(VI) concentration indicated that the model could predict pH-dependent U(VI) adsorption to within a root mean square error of 0.163-0.408, even under conditions outside of those for which the model was developed. These results indicate that this model could be used as a first approximation in predicting U(VI) adsorption and transport in the subsurface. U(VI) adsorption also decreased at pH >10, even in the absence of carbonate, which is of potential importance to U(VI) mobility in extreme environments present in the subsurface at some DOE facilities. The pH-dependent adsorption of U(VI) was fundamentally different in systems with a constant CO2 partial pressure as compared to a constant total carbonate concentration. Experiments at constant CO2 partial pressure may not be representative of the conditions present in the subsurface, and a constant carbonate concentration does not always result in decreased U(VI) adsorption at higher pH values. PMID- 11918021 TI - U.S. food agencies sponsor dioxin study. PMID- 11918022 TI - Antimony speciation in alkaline sulfide solutions: role of zerovalent sulfur. AB - Antimony is subject to a lower drinking water standard than arsenic, its notorious group 15 cohort in the periodic table. Both elements often co-vary in nature and are fairly soluble under reducing, alkaline conditions. Of the two, much less is known about the environmental chemistry of Sb. Measurements of Sb solubility in sulfidic solutions equilibrated with stibnite (Sb2S3) and orthorhombic sulfur reveal the existence of two new complexes that may control Sb behavior in many reducing environments. Formation reactions and stability constants (23 +/- 2 degrees C) are HS- + S(s) + Sb2S3(s) <==> HSb2S5-, log K = 1.47 +/- 0.17; and HS- + 2S(s) + Sb2S3(s) <==> Sb2S6(2-) + H+, log K = -9.55 +/- 0.07. The first complex is a mixed-valence Sb(III,V) complex; the second is an Sb(V) complex. Their stability in sulfidic solutions may explain previously puzzling evidence of Sb(V) in natural anoxic environments. Owing to these complexes, zerovalent S can enhance stibnite solubility up to 3 orders of magnitude. In neutral-to-alkaline, reducing environments, less than 7 microM HS- will transform O-coordinated, electrically neutral Sb(OH)3o to predominantly anionic S-coordinated complexes. This transformation could diminish the adsorption of Sb to negatively charged mineral surfaces, lowering retardation factors in anoxic aquifers. PMID- 11918023 TI - UV-irradiated DNA matrixes selectively bind endocrine disruptors with a planar structure. AB - DNA has unique chemical properties and is readily purified from salmon milts and shellfish gonads. DNA has few commercial uses and is generally discarded as an industrial waste. Recently, we prepared water-insoluble and nuclease-resistant DNA-films by UV irradiation. The DNA-films removed DNA-intercalating compounds. Here, we immobilized double-stranded DNA onto porous glass beads by UV irradiation and prepared DNA-immobilized glass bead columns. The DNA-immobilized columns effectively accumulated more DNA-intercalating materials than the DNA films. The DNA-immobilized columns bound endocrine disruptors with a planar structure, such as dioxin-derivatives, polychlorobiphenyl (PCB)-derivatives, and benzo[a]pyrene. Bisphenol A and diethylstilbestrol, which lack a planar structure, did not bind to the DNA-immobilized columns. These results suggest that DNA-immobilized glass bead columns could selectively remove chemical compounds with a planar structure by intercalation into the double-stranded DNA. DNA-immobilized glass beads and DNA-films prepared by UV irradiation have the potential to serve as useful biomaterials for medical, engineering, and environmental applications. PMID- 11918024 TI - Are national water quality assessments threatened? PMID- 11918025 TI - Evidence for nonlinear binding of PAHs to dissolved humic acids. AB - Binding of pyrene, fluoranthene, and phenanthrene to dissolved humic acids (HA) was determined by the fluorescence quenching (FQ) and complexation-flocculation (CF) methods. Determinations by the CF method, using varying contaminant concentrations and a constant HA concentration, yielded nonlinear Freundlich-type isotherms (n = 0.65-0.84). Experiments using both the CF and the FQ methods with varying HA concentrations and a constant contaminant concentration yielded curved "Stern-Volmer"-type plots that also indicate nonlinear binding. These findings suggest that linear partitioning or site complexation in the presence of excess available sites cannot fully describe the interactions of hydrophobic compounds with dissolved humic material. Site-specific hydrophobic interactions at limited interior or external molecular surfaces may be considered. PMID- 11918026 TI - Mold growth linked to airtight building designs. PMID- 11918027 TI - Arsenic accumulation and metabolism in rice (Oryza sativa L.). AB - The use of arsenic (As) contaminated groundwater for irrigation of crops has resulted in elevated concentrations of arsenic in agricultural soils in Bangladesh, West Bengal (India), and elsewhere. Paddy rice (Oryza sativa L.) is the main agricultural crop grown in the arsenic-affected areas of Bangladesh. There is, therefore, concern regarding accumulation of arsenic in rice grown those soils. A greenhouse study was conducted to examine the effects of arsenic contaminated irrigation water on the growth of rice and uptake and speciation of arsenic. Treatments of the greenhouse experiment consisted of two phosphate doses and seven different arsenate concentrations ranging from 0 to 8 mg of As L(-1) applied regularly throughout the 170-day post-transplantation growing period until plants were ready for harvesting. Increasing the concentration of arsenate in irrigation water significantly decreased plant height, grain yield, the number of filled grains, grain weight, and root biomass, while the arsenic concentrations in root, straw, and rice husk increased significantly. Concentrations of arsenic in rice grain did not exceed the food hygiene concentration limit (1.0 mg of As kg(-1) dry weight). The concentrations of arsenic in rice straw (up to 91.8 mg kg(-1) for the highest As treatment) were of the same order of magnitude as root arsenic concentrations (up to 107.5 mg kg( 1)), suggesting that arsenic can be readily translocated to the shoot. While not covered by food hygiene regulations, rice straw is used as cattle feed in many countries including Bangladesh. The high arsenic concentrations may have the potential for adverse health effects on the cattle and an increase of arsenic exposure in humans via the plant-animal-human pathway. Arsenic concentrations in rice plant parts except husk were not affected by application of phosphate. As the concentration of arsenic in the rice grain was low, arsenic speciation was performed only on rice straw to predict the risk associated with feeding contaminated straw to the cattle. Speciation of arsenic in tissues (using HPLC ICP-MS) revealed that the predominant species present in straw was arsenate followed by arsenite and dimethylarsinic acid (DMAA). As DMAA is only present at low concentrations, it is unlikely this will greatly alter the toxicity of arsenic present in rice. PMID- 11918028 TI - Physicochemical aspects of lead bioaccumulation by Chlorella vulgaris. AB - The relationship between lead speciation and its bioaccumulation by the alga Chlorella vulgaris was studied in the absence and presence of nitrilotriacetic, iminodiacetic, malonic, and citric acids. Pb uptake fluxes were rigorously analyzed by considering the simultaneous effects of metal transport in the medium coupled with metal complex dissociation kinetics. Under the conditions examined here, lead biouptake by C. vulgaris was governed by the free lead ion activity. Potentially labile hydrophilic complexes such as lead citrate and lead malonate did not contribute to the internalization fluxes. Kinetic modeling of the mass transport, adsorption reactions, and internalization fluxes confirmed the rate limiting role of the internalization flux. Comparison of the internalization and diffusive fluxes revealed that even in the presence of a large excess of Pb complexes, the supply of free ion (Pb2+) was sufficient to account for the observed Pb uptake. Pb adsorption to the cell surface was described by Langmuir isotherm. A new method was proposed as a means to estimate the number of Pb occupied transport sites at steady state. The apparent stability constant for the interaction of Pb with transport sites was determined to be 10(5.5) M(-1) at pH 6. Low temperature decreased both the Pb uptake flux and the Pb adsorbed to the transport sites. Pb uptake in the presence of Ca was competitively inhibited, and the binding affinity constant for Ca and transport sites was estimated to be 10(4.67) M(-1) at pH 6. Results were discussed within the perspective of the free ion activity and biotic ligand models. PMID- 11918029 TI - Arsenic(III) oxidation and arsenic(V) adsorption reactions on synthetic birnessite. AB - The oxidation of arsenite (As(III)) by manganese oxide is an important reaction in both the natural cycling of As and the development of remediation technology for lowering the concentration of dissolved As(III) in drinking water. This study used both a conventional stirred reaction apparatus and extended X-ray absorption fine structure (EXAFS) spectroscopy to investigate the reactions of As(III) and As(V) with synthetic birnessite (MnO2). Stirred reactor experiments indicate that As(III) is oxidized by MnO2 followed by the adsorption of the As(V) reaction product on the MnO2 solid phase. The As(V)-Mn interatomic distance determined by EXAFS analysis for both As(III)- and As(V)-treated MnO2 was 3.22 A, giving evidence for the formation of As(V) adsorption complexes on MnO2 crystallite surfaces. The most likely As(V)-MnO2 complex is a bidentate binuclear corner sharing (bridged) complex occurring at MnO2 crystallite edges and interlayer domains. In the As(III)-treated MnO2 systems, reductive dissolution of the MnO2 solid during the oxidation of As(III) caused an increase in the adsorption of As(V) when compared with As(V)-treated MnO2. This suggested that As(III) oxidation caused a surface alteration, creating fresh reaction sites for As(V) on MnO2 surfaces. PMID- 11918031 TI - Greening organic chemistry. ((())). PMID- 11918030 TI - Uptake and efflux of Cd and Zn by the green mussel Perna viridis after metal preexposure. AB - Cadmium and zinc uptake from the dissolved phase, assimilation efficiency from the dietary phase, efflux rate constants, and body burden as well as clearance rate were measured in the green mussel Perna viridis with or without laboratory preexposure to Cd or Zn. Efflux rate constants and clearance rates were little affected by preexposure to either Cd or Zn. In contrast, the assimilation of Cd increased by 1.2-1.6 in mussels preexposed to Cd (subsequent Cd concentrations 10.2-25.9 microg(-1)) as compared to controls (0.19-0.39 microg g(-1)). This increase corresponded to an elevation in the proportion of Cd associated with the metallothionein-like proteins (MTLPs) in the mussels, suggesting that exposure to Cd and subsequent induction of MTLPs affected Cd accumulation. Exposure to Zn only resulted in elevated body concentrations following 7-d exposure to 250 microg L(-1), although Zn and Cd uptake from the dissolved phase were reduced by 24-47% by exposure to a lower concentration (100 microg L(-1)) for 7 and 21 d. Despite the lack of an increase in body Zn concentration, the subcellular distribution was altered such that the proportion of Zn associated with the metal rich granules increased. This study indicates the importance of the subcellular distribution of metals in affecting the biokinetics and thus the toxic effects of metals on aquatic animals. Cd preexposure has potential effects on its influx from the dietary phase, e.g., increasing the importance of dietary uptake and further increasing the body burdens. In contrast, preexposure to Zn has a negative effect on Cd and Zn influx from the dissolved phase, suggesting the mechanism of Zn regulation but also potentially reducing Cd uptake and body concentrations over the long-term exposure. Such effects may have implications for biomonitoring studies involving a single species that modifies physiological processes affecting metal uptake (and hence bioavailability). Caution is needed in extrapolating data to species not capable of making such changes, particularly for Cd, which is not regulated and for which the effects of an elevated body burden are most obvious. PMID- 11918032 TI - Medical errors and wrong-site surgery. PMID- 11918033 TI - Radiologic case study. Failed anterior cruciate ligament (ACL) reconstruction. PMID- 11918034 TI - Broken guide pin: a method of retrieval from within the hip joint. AB - Broken guide pin removal from within the hip joint can be accomplished using a combined method of drilling and suction. This technique also may be of value when a small loose body or foreign object needs to be retrieved from the hip joint. PMID- 11918035 TI - Hyperbaric oxygen. AB - This cutting edge article discusses the most frequent uses of hyperbaric oxygen for the orthopedic surgeon. Hyperbaric oxygen therapy is an adjunct to orthopedic interventions when healing problems are anticipated due to wound hypoxia or uncontrolled infection. PMID- 11918036 TI - Use of injectable calcium phosphate in the treatment of intra-articular distal radius fractures. AB - This study reviewed 19 patients with intra-articular fractures of the distal end ofthe radius treated with augmented external fixation and injectable calcium phosphate (Norian SRS; Norian Corp, Cupertino, Calif) between November 1997 and July 2000. Average patient age was 52 years, and average follow-up was 16.3 months. Patients were evaluated clinically and radiographically using the modified Mayo Wrist Score and Knirk and Jupiter's criteria, respectively. Overall results are comparable to those from other studies of intra-articular distal radius fractures in which other grafts have been used. PMID- 11918037 TI - Preoperative administration of epoetin alfa to total joint arthroplasty patients. AB - Epoetin alfa is indicated to reduce allogeneic transfusions in patients undergoing major operations. This study included 40 patients undergoing total joint arthroplasty who received 600 IU/kg doses of epoetin alfa 21, 14, and 7 days preoperatively. Statistically equal matches were obtained according to preoperative hemoglobin, operation, sex, and age. After controlling for autologous donation, the average hemoglobin level on postoperative day one in the treatment group (10.93 g/dL) was significantly higher than the matched patient group (9.86 g/dL). Likewise, treated patients were transfused with significantly less blood (0.68 units/patient) compared to the matched group (1.6 units/patient). In these patients, administration of epoetin alfa was associated with significantly higher perioperative hemoglobin levels and significantly fewer transfusions than matched counterparts. The use of epoetin alfa in this heterogeneous surgical population illustrates its success in a varied population. PMID- 11918038 TI - Posterior pelvic tilt in patients with decreased lumbar lordosis decreases acetabular femoral head covering. AB - The relationship between pelvic tilt and acetabular femoral head covering was determined in 38 patients with decreased lumbar lordosis. Posterior pelvic tilt and acetabular femoral head covering were measured on radiographs taken in both supine and standing positions. Previously reported normal values obtained from 96 patients were used as controls. Posterior pelvic tilt increased (P<.01) in patients with decreased lumbar lordosis and anterior acetabular femoral head covering decreased (P<.0001) compared with the controls. These differences were more remarkable in the standing position than in the supine position. Patients with decreased lumbar lordosis have decreased femoral head covering, especially in the standing position. PMID- 11918039 TI - Revision total knee arthroplasty using large distal femoral augments for severe metaphyseal bone deficiency: a preliminary study. AB - Managing severe structural femoral metaphyseal bone loss in revision total knee arthroplasty is a challenging problem facing the revision knee surgeon. This study assesses the use of large (30 mm) metal distal femoral augments to compensate for severe bone deficiencies. Hospital for Special Surgery scores, Knee Society scores, and range of motion improved after implantation of femoral components with 30-mm distal femoral augments. There was no radiographic evidence of loosening, and no implants had been revised at mean 37-month follow-up. This appears to be an acceptable technique based on the intermediate-term results. PMID- 11918040 TI - Bilateral proximal third fibular stress fractures in an adolescent female track athlete. AB - Proximal fibular stress fracture is an uncommon injury in athletes but has been frequently reported in the military population. Although the true incidence is unknown and its pathophysiology is not completely understood, proximal fibular stress fracture is believed to be a rare injury and awareness of it will help clinicians make the proper diagnosis and provide appropriate treatment. PMID- 11918041 TI - Squamous cell carcinoma arising from chronic osteomyelitis of the patella. PMID- 11918042 TI - The association between Charcot-Marie-Tooth disease and developmental dysplasia of the hip. PMID- 11918043 TI - Surgical treatment of symptomatic Bertolotti's syndrome in post-fusion patients. PMID- 11918044 TI - Arthroscopic anterior cruciate ligament reconstruction using patellar tendon autografts. PMID- 11918046 TI - TB in badgers and cattle: trying to find the answers. PMID- 11918045 TI - 2000 sees a rise in sales of antimicrobials used in food animals. PMID- 11918047 TI - Postal survey of ovine caseous lymphadenitis in the United Kingdom between 1990 and 1999. AB - Members of the Sheep Veterinary Society and three terminal sire breed societies (A, B and C) were surveyed to investigate the prevalence and characteristics of ovine caseous lymphadenitis in the UK. In total, 264 veterinary surgeons and 510 farmers responded. Eighteen per cent of the vets had seen at least one case of the disease, and 45 per cent of the farmers had seen abscesses in their sheep. Relatively few farmers had had the cause of the abscesses investigated, but 24 of the 32 flocks that had submitted samples to a laboratory had had the disease confirmed. The incidence of the disease and abscesses increased from 1990 to 1999. The disease was more likely to be investigated in certain breeds and when male sheep had abscesses. Risk factors for the disease included the presence of sheep of breed B in the flock, the presence of half-bred sheep, showing sheep, and the use of shearing contractors. Dipping sheep appeared to be protective. PMID- 11918048 TI - Serological prevalence of Mycoplasma bovis infection in suckling beef cattle in France. AB - The prevalence of Mycoplasma bovis infection in France was assessed by means of a serological survey of suckling beef cattle, using an ELISA. The survey included 824 randomly selected herds in eight French counties and a total of 32,197 animals more than one year old. In each county, the number of herds tested was determined statistically on the basis of the hypothesis that about 40 per cent of herds are infected. The proportion of herds containing at least one infected animal ranged from 28 to 90 per cent depending on the county. Among the 32,197 sera tested, the animal infection rate ranged between 2 per cent and 13 per cent. In infected herds, the average number of positive animals per herd was between 10 and 20 per cent, and the infection was unevenly distributed among the areas tested. PMID- 11918049 TI - Pyogranulomatous myocarditis due to Staphylococcus aureus septicaemia in two harbour porpoises (Phocoena phocoena). AB - Staphylococcus aureus septicaemia was diagnosed in a dead, stranded harbour porpoise from the German Baltic Sea and in a live harbour porpoise by-caught in inner Danish waters and taken into captivity. Lesions included pyogranulomatous myocarditis, necrotising suppurative bronchopneumonia, pyelonephritis, osteomyelitis and leptomeningitis, and abscesses in lymph nodes and skeletal muscles. The captive animal had fibrinous suppurative epicarditis and pyogranulomatous myocarditis with abscesses. In both animals the organism was suspected to have entered through skin lesions or via the respiratory tract. PMID- 11918050 TI - Outbreak of Marek's disease in a flock of turkeys in Scotland. PMID- 11918051 TI - Epileptiform convulsions in a cow with degeneration of the hippocampus. PMID- 11918052 TI - Use of ultrasonography in the diagnosis of hepatic spindle-cell sarcoma in a savannah monitor (Varanus exanthematicus). PMID- 11918053 TI - Testicular and epididymal weights in the domestic dog (Canis familiaris). PMID- 11918054 TI - Need for vigilance in the aftermath of FMD and CSF. PMID- 11918055 TI - Vets and the future of farming. PMID- 11918059 TI - Biologically active secondary metabolites from the ascomycete A111-95. 2. Structure elucidation. PMID- 11918058 TI - Isolation of a novel cyclic hexadepsipeptide pipalamycin from Streptomyces as an apoptosis-inducing agent. AB - The novel cyclic hexadepsipeptide named pipalamycin was isolated from a culture filtrate of Streptomyces sp. ML297-90F8 as an apoptosis-inducing agent. The antibiotic was found to be consisting of each one mole of alanine, N hydroxyalanine, glycine, N-acylated 3-hydroxyleucine, and two moles of piperazic acid. Pipalamycin induced apoptosis in apoptosis-resistant human pancreatic adenocarcinoma AsPC-1 cells at 0.3 microg/ml in 24 to approximately 48 hours. It also showed antibacterial activity on Gram-positive bacteria such as Staphylococcus aureus and Micrococcus luteus. Fermentation, isolation, structural elucidation and the biological activities of pipalamycin are described. PMID- 11918060 TI - A novel antibiotic CJ-17,572 from a fungus, Pezicula sp. AB - A new antibiotic, CJ-17,572 (I) was isolated from the fermentation broth of a fungus Pezicula sp. CL11877. The structure of I was determined to be a new equisetin derivative by spectroscopic analyses. The compound inhibits the growth of multi-drug resistant Staphylococcus aureus and Enterococcusfaecalis with IC50s of 10 and 20 microg/ml, respectively. PMID- 11918061 TI - CJ-21,058, a new SecA inhibitor isolated from a fungus. AB - A new equisetin derivative, CJ-21,058 (I) was isolated from the fermentation broth of an unidentified fungus CL47745. It shows antibacterial activity against Gram-positive multi-drug resistant bacteria by inhibiting ATP-dependent translocation of precursor proteins across a bacterial cell membrane. PMID- 11918062 TI - YM-181741, a novel benz[a]anthraquinone antibiotic with anti-Helicobacter pylori activity from Streptomyces sp. AB - A novel benz[a]anthraquinone, YM-181741, was isolated from the culture broth of actinomycete strain Q57219. The strain was identified as Streptomyces sp. by morphological and chemotaxonomic characterization. YM-181741 was purified from the culture supernatant by serial column chromatography. The structure of YM 181741 was determined by spectroscopic analysis including one- and two dimensional NMR experiments. YM-181741 showed selective anti-Helicobacterpylori activity with a MIC value of 0.2 microg/ml. PMID- 11918063 TI - Biologically active secondary metabolites from the ascomycete A111-95. 1. Production, isolation and biological activities. AB - Eight secondary metabolites were isolated from submerged cultures of the ascomycete A111-95 during a search for new nematicidal metabolites. (-) Galiellalactone (7) and compound 2 are metabolites previously obtained from cultures of Galiella rufa while the compounds 1, 3, 4, 5, 6 and 8 (3 and 4 were obtained as an unseparable mixture), were isolated as natural products for the first time. Compound 2, pregaliellalactone (5) and the mixture of 3 and 4 showed nematicidal activities towards Caenorhabditis elegans and Meloidogyne incognita. All compounds showed moderate or weak cytotoxic activities. PMID- 11918064 TI - Studies on the biosynthesis of epothilones: hydroxylation of Epo A and B to epothilones E and F. AB - When Sorangium cellulosum So ce90 is grown without XAD adsorber resin there is a steady state of epothilone A and B biosynthesis and hydroxylation of these products to epothilones E and F. This biotransformation at position C-21 of the thiazole ring is not restricted to producers of epothilones. It is carried out by a substrate induced monooxygenase. Epothilones E and F are further degraded by opening of the lactone ring by an esterase. Steps of degradation of different strains of S. cellulosum are compared. PMID- 11918065 TI - Biosynthesis of hibarimicins. I. 13C-labeling experiments. AB - Biosynthesis of hibarimicin was studied based on the feeding experiments with 13C labeled acetates. All carbons in the aglycon, except for the methoxy carbons, were derived from acetate. The carbon framework of the aglycon was proved to be constructed by dimerization of an intermediate which was biosynthesized via the decarboxylation and skeltal rearrangement starting from an undecaketide. The rearrangement was confirmed by detecting the long range (three-bond) coupling between two carbons in the difference spectra of selective 13C decoupled INADEQUATE of hibarimicin B labeled with sodium [1,2-13C2] acetate. PMID- 11918066 TI - Biosynthesis of hibarimicins. II. Elucidation of biosynthetic pathway by cosynthesis using blocked mutants. AB - The biosynthetic pathway of hibarimicin (HBM) was proposed on the basis of the experimental results obtained by using blocked mutants of Microbispora rosea subsp. hibaria TP-A0121, the HBM producer. In its biosynthesis, the oxidative coupling of the aromatic undecaketide unit generates a symmetrical aglycon HMP-Y1 (hibarimicin-mutant product Y1), which is oxidatively modified to hibarimicinone, the HBM aglycon. The following glycosylation of hibarimicinone gives rise to the HBM complex. We identified that HMP-Y1 prepared by methanolysis of HMP-Y6, a glycosylated metabolite from a blocked mutant, was the key intermediate: transformation of 13C-labeled HMP-Y1 to HBM B was confirmed by NMR measurements. Mutant strain produced another type of aglycon HMP-P1 in which the coupled polyketide units were intramolecularly bridged by the ether bond. This metabolite also arose by the spontaneous elimination of methanol molecule from hibarimicinone. PMID- 11918067 TI - CJ-15,696 and its analogs, new furopyridine antibiotics from the fungus Cladobotryum varium: fermentation, isolation, structural elucidation, biotransformation and antibacterial activities. AB - CJ-15,696 and 7 novel furopyridine antibiotics were isolated from the fungus Cladobotryum varium CL12284. Their structures were determined by X-ray crystallography and spectral analysis. Three biotransformed analogs were also prepared from CJ-15,696. CJ-15,696 showed moderate activity against various Gram positive bacteria including some drug resistant strains such as methicillin resistant Staphylococcus aureus (MRSA). PMID- 11918068 TI - Biosynthesis of hibarimicins. III. Structures of new hibarimicin-related metabolites produced by blocked mutants. AB - Structures of metabolites produced by blocked mutants of Microbispora rosea subsp. hibaria TP-A0121, hibarimicin-producer, were determined by spectroscopic analysis. HMP-Y6 is the dimer of the west half of hibarimicin B, the aglycon of which is the genuine biosynthetic intermediate. HMP-P1 is the shunt product arising from the release of a methanol molecule from hibarimicinone. HMP-P4, the glycoside of HMP-P1, is glycosylated with two amicetoses and two digitoxoses same as hibarimicin B. HMP-M1, M2, M3 and M4 are shunt products derived from the monomeric undecaketide intermediates. PMID- 11918069 TI - SNF4435C and D, novel immunosuppressants produced by a strain of Streptomyces spectabilis. III. Immunosuppressive efficacy. AB - Novel immunosuppressants, SNF4435C and D produced by a strain of Streptomyces spectabilis, were examined for their pharmacodynamical profiles. SNF4435C and D suppressed the responses of both murine splenocytes and human peripheral blood lymphocytes in the mixed lymphocyte reaction (MLR) with IC50 values of 0.5 microM and 0.2 microM, respectively. In the mouse MLR experiments, SNF4435C and D did not block the production of interleukin-2 (IL-2) and the compounds-induced suppression was not restored by the addition of exogeneous IL-2. In addition, the significant inhibitory action was still retained even when SNF4435C or D was added after 48 hours from the start of the culture. These results were distinct from the behaviors observed with FK-506. SNF4435C, the major component, suppressed mouse delayed type hypersensitivity (DTH) and prolonged rat skin allograft survival. PMID- 11918070 TI - Enhancement of fibrinolytic activity of U937 cells by malformin A1. AB - We have found that malformin A1, a cyclopentapeptide metabolite of Aspergillus niger, enhanced 2.0- to 3.2-fold the 125I-fibrin clot lysis when incubated at 1 to approximately 10 microM with both U937 cells and blood plasma, both of which were essential to the malformin A1 action. The effect was inhibited by epsilon aminocaproic acid and anti-urokinase serum, but not by anti-tissue-type plasminogen activator IgG, showing that the enhancement was mediated by urokinase catalyzed plasminogen activation. However, malformin A1 affected neither cellular urokinase activity nor cell-free reactions involved in the fibrinolytic pathway. Malformin-treated, washed cell had an increased capacity to degrade fibrin in the presence of plasma. These results suggest that malformin A1 enhances fibrinolytic activity by affecting cell-mediated response to initiate and/or propagate fibrinolytic activity. PMID- 11918071 TI - Antibiotic A10255 (thioplabin) enhances fibrin binding and activation of plasminogen. AB - Three thiopeptide metabolites that enhance fibrin binding of plasminogen were isolated from a culture of Streptomyces sp. R1401. A combination of spectroscopic analyses revealed that these compounds were identical with the antibiotic A10255B, E and G. These agents enhanced fibrin binding of plasminogen and plasminogen/urokinase-mediated fibirinolysis at concentrations of 5 to approximately 20 microM. A10255B reversibily increased urokinase-catalyzed activation of plasminogen by lowering Km, while the agent did not enhance urokinase activity when substrates other than plasminogen were used, indicating that the agent affects plasminogen to increase its affinity to urokinase. A smaller but significant increase in activation was also observed when conformationally relaxed plasminogen derivatives such as Lys-plasminogen and mini plasminogen were used. Two related thiopeptide antibiotics with a C-terminal amide had no effect on plasminogen activation, suggesting a role of the terminal carboxyl of the A10255 molecule in activity. PMID- 11918072 TI - Synthesis and activities of bactobolin derivatives having new functionality at C 3. AB - Some derivatives of bactobolin were prepared from bactobolin (1) by transformation of the dichloromethyl group at C-3 to the hydroxymethyl, carboxylic acid, methanesulfonyloxymethyl and aldehydeoxime groups. The derivatives proved to be less active than the parent antibiotic 1 against bacteria as well as cytotoxicity, indicating that the functionality at C-3 considerably influences the biological activity. PMID- 11918073 TI - Inhibition of recombinant sphingosine kinases by novel inhibitors of microbial origin, F-12509A and B-5354c. PMID- 11918074 TI - A comparative docking study and the design of potentially selective MMP inhibitors. AB - As part of a program aimed at the design and synthesis of constrained MMP inhibitors, a survey of the reported X-ray and NMR structures of MMP/inhibitor complexes was performed, revealing mutations of key amino acids at different subsites between MMPs. A comparative study of fully automated docking programs AutoDock and DOCK in closely approximating the X-ray crystal structures of ten selected MMP inhibitors was performed. AutoDock proved to be highly reliable, efficient and predictive for a set of inhibitors with less than six atom types. PMID- 11918075 TI - High-affinity interactions of ligands at recombinant guinea pig 5HT7 receptors. AB - The serotonin 5HT7 receptor has been implicated in numerous physiological and pathological processes from circadian rhythms to depression and schizophrenia. Clonal cell lines heterologously expressing recombinant receptors offer good models for understanding drug-receptor interactions and development of quantitative structure-activity relationships (QSAR). Comparative Molecular Field Analysis (CoMFA) is an important modern QSAR procedure that relates the steric and electrostatic fields of a set of aligned compounds to affinity. Here, we utilized CoMFA to predict affinity for a number of high-affinity ligands at the recombinant guinea pig 5HT7 receptor. Using R-lisuride as the template, a final CoMFA model was derived using procedures similar to those of our recent papers. The final cross-validated model accounted for >85% of the variance in the compound affinity data, while the final non-cross validated model accounted for >99% of the variance. Model evaluation was done using cross-validation methods with groups of 5 ligands. Twenty cross-validation runs yielded an average predictive r2(q2) of 0.779 +/- 0.015 (range: 0.669-0.867). Furthermore, 3D chemical database search queries derived from the model yielded hit lists of promising agents with high structural similarity to the template. Together, these results suggest a possible basis for high-affinity drug action at 5HT7 receptors. PMID- 11918077 TI - Functional group placement in protein binding sites: a comparison of GRID and MCSS. AB - One approach to combinatorial ligand design begins by determining optimal locations (i.e., local potential energy minima) for functional groups in the binding site of a target macromolecule. MCSS and GRID are two methods, based on significantly different algorithms, which are used for this purpose. A comparison of the two methods for the same functional groups is reported. Calculations were performed for nonpolar and polar functional groups in the internal hydrophobic pocket of the poliovirus capsid protein, and on the binding surface of the src SH3 domain. The two approaches are shown to agree qualitatively; i.e., the global characteristics of the functional group maps generated by MCSS and GRID are similar. However, there are significant differences in the relative interaction energies of the two sets of minima, a consequence of the different functional form used to evaluate polar interactions (electrostatics and hydrogen bonding) in the two methods. The single sphere representation used by GRID affords only positional information, supplemented by the identification of hydrogen bonding interactions. By contrast, the multi-atom representation of most MCSS groups yields in both positional and orientational information. The two methods are most similar for small functional groups, while for larger functional groups MCSS yields results consistent with GRID but superior in detail. These results are in accord with the somewhat different purposes for which the two methods were developed. GRID has been used mainly to introduce functionalities at specific positions in lead compounds, in which case the orientation is predetermined by the structure of the latter. The orientational information provided by MCSS is important for its use in the de novo design of large, multi-functional ligands, as well as for improving lead compounds. PMID- 11918076 TI - A genetic algorithm for structure-based de novo design. AB - Genetic algorithms have properties which make them attractive in de novo drug design. Like other de novo design programs, genetic algorithms require a method to reduce the enormous search space of possible compounds. Most often this is done using information from known ligands. We have developed the ADAPT program, a genetic algorithm which uses molecular interactions evaluated with docking calculations as a fitness function to reduce the search space. ADAPT does not require information about known ligands. The program takes an initial set of compounds and iteratively builds new compounds based on the fitness scores of the previous set of compounds. We describe the particulars of the ADAPT algorithm and its application to three well-studied target systems. We also show that the strategies of enhanced local sampling and re-introducing diversity to the compound population during the design cycle provide better results than conventional genetic algorithm protocols. PMID- 11918078 TI - Differential expression of an alpha-galactosyl-containing trisaccharide on high- and low-malignant murine sarcoma cells: identification and regulation. AB - Past studies have shown that carbohydrate residues reactive with the Griffonia simplicifolia isolectin B4 (GS I-B4) are present on the surface of highly malignant murine sarcoma cells but are lacking or expressed in much lower amounts on the surface of low-malignant cells isolated from the same parent tumors (Am J Pathol 111: 27; J Nat Cancer Inst 71: 1281). In the present study it is shown that an antibody which recognizes the trisaccharide Galalpha1-3Galbeta1-4GlcNAc- is reactive with the highly-malignant cells but is non-reactive with the low malignant cells. Further studies show that the high-malignant cells not only bind GS 1-B4 but also bind Evonymus europaea lectin (which like GS I-B4 recognizes terminal galactose in alpha1-3 linkage) and Erythina crystagalli lectin (which recognizes sub-terminal galactose in the beta1-4 linkage--e.g., Galbeta1 4GlcNAc). In contrast, the low malignant cells bind Erythina crystagalli lectin as efficiently as the high malignant cells but do not bind (or bind much smaller amounts of) either GS I-B4 or Evonymus europaea lectin. The present studies also show that there is no significant difference between high- and low-malignant cells in expression of alpha-galactosidase activity. In contrast, the high malignant cells express high levels of alpha-galactosyl transferase activity while this enzyme is virtually undetectable in low-malignant cells. Taken together, these studies indicate that differential expression of a single monosaccharide residue distinguishes high- and low-malignant murine sarcoma cells. These studies also identify a mechanism to account for surface carbohydrate differences between the high- and low-malignant cells. PMID- 11918079 TI - Differential experimental micrometastasis to lung, liver, and bone with lacZ tagged CWR22R prostate carcinoma cells. AB - LacZ-tagged human prostate carcinoma CWR22Rv1 cells metastasize spontaneously to lung, liver, and bone from subcutaneous primary tumors in athymic nude mice; these organs are 'natural' targets of metastasis for the human disease. To evaluate the mechanism(s) of metastasis to these organs, an experimental metastasis model was used by taking advantage of the ultrasensitive detection of lacZ. Within I h after tail vein injection, micrometastases were forming in lung, liver, bone, kidney, and brain with very different quantitative levels. The kinetics of loss of unstable micrometastases and retention of stable ones were also very different in these organs. After injecting suspensions of single cells, both whole-organ and serial-section staining for lacZ revealed considerable heterogeneity in cell number of individual lung micrometastases while micrometastases in liver contained only I or 2 cells. The size of individual bone micrometastases also suggested only 1 or 2 cells. Tumor cells could also be detected in the small blood vessels of the lung within minutes after injection. These studies indicate that lacZ-tagged CWR22Rv1 cells after tissue culturing contain subsets of cells capable of establishing transient micrometastases in lung, liver, and bone after direct injection into the animal's circulation. Moreover, the quantitative and qualitative properties of the micrometastases in the three organs differ significantly, suggesting different mechanisms for stabilization and fates of micrometastases in these organs. PMID- 11918080 TI - The regulation of prostate cancer cell adhesion to human bone marrow endothelial cell monolayers by androgen dihydrotestosterone and cytokines. AB - A previous study from our laboratory suggested that prostate cancer metastasis to bone may be mediated, in part, by preferential adhesion to human bone marrow endothelial (HBME) cells. Tumor cell adhesion to endothelial cells may be modulated by the effect of cytokines on cell adhesion molecules (CAMs). Tumor necrosis factor-alpha (TNF-alpha) regulates VCAM expression on the endothelium and this effect is enhanced by dihydrotestosterone (DHT). Transforming growth factor-beta (TGF-beta) stimulates the expression of alpha2beta1 integrin on PC-3 cells. The current study investigated the effects of the above cytokines and DHT (singularly and in various combinations) upon HBME and prostate cancer cell expression of VCAM, alpha2 integrin subunit, and beta1 integrin subunit by flow cytometry. We also monitored the effects of the above treatments on PC-3 cell adhesion to HBME monolayers. The data demonstrate that none of the treatments significantly altered the expression of selected CAMs on HBME cell and neoplastic prostate cell lines. The treatment of HBME monolayers with various combinations of cytokines and DHT prior to performing adhesion assays with PC-3 demonstrates that treatments containing TGF-beta reduced PC-3 cell adhesion to HBME monolayers by 32% or greater (P < 0.05). The reduction in PC-3 cell adhesion to TGF-beta treated HBME monolayers was dose dependent. Interestingly, LNCaP cells but not PC 3 cells treated with TGF-beta had a reduced ability to adhere to untreated HBME monolayers. These results suggest that TGF-beta may reduce tumor cell adhesion to bone marrow microvascular endothelium, in vivo. The biological significance of this observation is discussed. PMID- 11918081 TI - MMTV-associated transcription factor binding sites increase nm23-H1 metastasis suppressor gene expression in human breast carcinoma cell lines. AB - We hypothesize that elevation of nm23-HI metastasis suppressor gene expression in micrometastatic tumor cells may reduce their subsequent colonization and invasion, and induce differentiation, with a clinical benefit. This report presents the first analysis of the nm23-HI promoter to identify sites which can increase its transcription. Deletion mapping of a 2.1 kb nm23-H1 promoter fragment tethered to a reporter gene identified three regions involved in its differential expression levels among a panel of human breast carcinoma cell lines: a 195 bp NheI-XbaI fragment responsible for basal expression levels, a 248 bp AvrII-Nhel fragment which contributed to the elevated nm23-H1 expression observed in the high expressing cell lines, and a 544 bp AvrII fragment containing an inhibitory element. Examination of the 248 bp AvrII-NheI fragment revealed the unexpected presence of three transcription factor binding sites (MAF/Ets, CTF/NF1 half site and ACAAAG enhancer) previously identified in the MMTV-LTR, and in WAP and milk gene promoters, proposed to mediate mammary specific gene expression. Mutation of the three sites, individually or together, resulted in two-fold reductions in reporter gene expression. As controls, the same panel of mutations caused a different pattern of reporter gene expression in a non-mammary cell line, and mutation of another nearby site was without effect on nm23-HI. Our data identify a complex regulatory pattern for nm23-H1 transcription, and suggest that a mammary-specific cassette of transcription factors contribute to its elevated expression PMID- 11918082 TI - Synergistic effect of interleukin-2 and a vaccine of irradiated melanoma cells transfected to secrete staphylococcal enterotoxin A. AB - We have previously reported that immunization of mice with melanoma cells transfected to secrete the superantigen, Staphylococcal enterotoxin A (SEA), increased the production of antibodies to the B700 melanoma antigen, stimulated the production of endogenous interleukin 2 (IL-2), activated the expression of CD4, CD8 and CD25 T cell markers and enhanced NK cell activity. Now we show that immunization of mice with a vaccine of irradiated sea-transfected melanoma cells coupled with IL-2 therapy was even more effective in inhibiting the growth of primary melanoma tumors and the development of lung metastases than was the irradiated melanoma cell vaccine alone or IL-2 alone. The morphological and immunological effectiveness of the therapy was dose-dependent on IL-2. PMID- 11918083 TI - Stromal extracellular matrix reduces chemotherapy-induced apoptosis in colon cancer cell lines. AB - Several studies have shown that extracellular matrix reduces chemotherapeutic drugs-induced apoptosis in small cell lung cancer cells, myelomas and gliomas. We have investigated the protective effect of defined extracellular matrix components and of extracellular matrix from different cell types (fibroblasts, hepatocytes and intestinal epithelial cells) on the toxicity of three types of chemotherapeutic drugs on colon cancer cells. Human colon cancer cell lines LS174T and LiM6 were plated on plastic, on hepatocyte-derived ECM or on stromal ECM and in the presence of the antimetabolite 5-fluorouracil (5-FU). the topoisomerase I inhibitor camptothecin and the topoisomerase II inhibitor etoposide. We determined IC50 for the drugs for each of these culture conditions. We also determined the expression of the anti-apoptotic proteins bcl-2 and bcl-x (L) under these culture conditions. We found that stromal ECM protected LiM6 cells from the toxicity of etoposide and LS174T, but not LiM6 cells, from the toxicity of camptothecin. Collagen 1, fibronectin and fibroblast-derived ECM rendered LiM6 cells, but not LS174T, more sensitive to the harmful effect of 5 FU. Both colon cell lines had increased expression of anti-apoptotic proteins bcl 2 and bcl-x(L) when cultured on the various ECMs and with the drugs, but there was no correlation between a protective ECM effect and expression of the anti apoptotic proteins. Stromal-derived ECM may protect colon cancer cells from etoposide and camptothecin-induced apotosis, through a mechanism that is not bcl 2 or bcl-x(L) dependant. PMID- 11918084 TI - Carbohydrate expression profile of colorectal cancer cells is relevant to metastatic pattern and prognosis. AB - Carbohydrate expression of cancer cells is closely related to the metastatic nature of colorectal cancer. In the present study we investigated the relevance of carbohydrate expression profiles of colorectal cancer cells in the primary lesion to metastatic distribution patterns as well as prognosis in 134 cases. Carbohydrate expression was estimated by histochemistry with 17 kinds of lectins and 3 kinds of Lewis-related monoclonal antibodies (MAbs), and correlations between the staining and clinicopathological parameters were examined. The results showed that lymphatic invasion, lymph node metastasis, and peritoneal metastasis correlated with staining with lectins that bind galactose/N acetylgalactosamine residues (Gal/GalNAc) such as Maclura pomifera (MPA), Arachis hypogaea (PNA), Helix pomatia (HPA), and Vicia villosa (VVA). In contrast, hepatic metastasis correlated with staining with Anguilla anguilla lectin (AAA), anti-LewisX (LEX-2), anti-sialyl Lewisa (NS 19-9), and anti-sialyl-dimeric LewisX (FH-6) MAbs, all of which bind preferentially to fucosylated carbohydrate chains. The five-year survival rate of patients was related to the staining of cancers with MPA, HPA, FH-6 or NS19-9, and MPA- and FH-6 staining were independent prognostic factors. We conclude that carbohydrate expression profiles of cancer cells are relevant to the route of tumor cell dissemination, metastatic pattern as well as prognosis of colorectal cancer. PMID- 11918085 TI - Is reduced expression of mismatch repair genes MLH1 and MSH2 in patients with sporadic colorectal cancer related to their prognosis? AB - The majority of mutations in hereditary nonpolyposis colon carcinoma (HNPCC) patients affect the mismatch-repair genes (MMRG) MLHI and MSH2. In addition, mutations of these genes were found in about 15% of sporadic colorectal carcinomas which appear to be related to microsatellite instability (MSI). However, mutations in MMRG were not found in all MSI-positive carcinomas, but MMRG mutations may be relevant for the assessment of tumor characteristics and patients' prognosis. Therefore, we investigated the relationship between expression of MMRG, tumor biology and patients' survival. In 127 patients with sporadic colorectal carcinomas and a minimum of 5 years follow-up after curative surgery immunohistochemical detection of MLHI and MSH2 was analyzed semiquantitatively. Lost expression of MLHI has been found in tumor specimens from 10 patients, whereas MSH2 expression was missing in 5 patients. This reduced expression did not correlate with tumor stage, lymph node involvement, grading or tumor invasion into blood vessels. However, a significant correlation was found for lymphovascular invasion (P = 0.02) and localization within the colorectum (P = 0.003) in MLH1-negative carcinomas. In addition, although there was a clear tendency for longer overall survival (72 vs. 63 months) for patients with MLH1 negative carcinomas, significant differences for overall and recurrence-free survival were not seen. In conclusion of our results and a critical review of literature, the prognostic importance of the MMR genes in sporadic colorectal carcinomas remains controversial. PMID- 11918086 TI - An inhibitor of stress-activated MAP-kinases reduces invasion and MMP-2 expression of malignant melanoma cells. AB - A lot of parallels have been described between invasion of malignant tumor cells and leukocyte movement during inflammatory responses. Concerning these similarities, we investigated the function of cytokine-suppressive anti inflammatory drugs (CSAIDs), which act via inhibition of stress-activated MAP kinases, in regulation of expression of proteolytic enzymes and in vitro invasion of malignant melanoma cells. The p38MAPK inhibitor SB203580 reduced matrigel invasion of MeWo cells by 60%, while the MEK-1 inhibitor PD98059 did not have any effect on invasion. Active p38MAPK was detected in MeWo cells by immunoblotting and confocal microscopy. Cells showed a constitutive expression of matrix metalloproteinase (MMP)-2 as well as tissue inhibitor of metalloproteinases (TIMP)-1 and TIMP-2 mRNAs. Expression of MMP-1 or urokinase-type plasminogen activator (uPA) was not detected by Northern blot. Inhibition of p38MAPK by the specific inhibitor SB203580 resulted in downregulation of MMP-2 mRNA and protein levels as well as gelatinolytic activity, while expression levels of TIMP-1 and TIMP-2 mRNAs were not changed. The specific MEK-1 inhibitor PD98059 did not change expression of MMP-2 or TIMPs. Neither SB203580 nor PD98059 changed proliferation of cells. The results suggest that stress-activated protein kinases like p38MAPK are involved in regulation of expression of MMP-2 as well as in vitro invasion of malignant melanoma cells. Inhibitors of p38MAPK may be promising substances to interfere with a signaling cascade associated with invasion of malignant tumor cells. PMID- 11918087 TI - Monocyte chemoattractant protein-1 (MCP-1), secreted by bone marrow endothelial cells, induces chemoattraction of 5T multiple myeloma cells. AB - Homing of multiple myeloma (MM) cells to the bone marrow (BM) requires transendothelial migration. In the present work we tested whether monocyte chemoattractant protein-1 (MCP-1) and CCR2, the high affinity receptor for MCP-1, are involved in this process. Murine 5T2 and 5T33MM cell lines were selected as source of MM cells and STR4, 10 and 12 of BM endothelial cells (BMEC). RT-PCR demonstrated transcripts for MCP-1 in BMEC and ELISA the presence of MCP-1 protein in culture medium. RNase protection assay showed mRNA expression for CCR2, and FACS analysis the presence of CCR2 protein on the MM cells. EC conditioned medium induced chemoattraction of MM cells, a phenomenon inhibited by anti-MCP-1 antibodies. In conclusion, MM cells express CCR2 and are attracted by MCP-1 secreted by BMEC. We suggest that local MCP-1 production by BMEC is one of the mechanisms involved in homing of myeloma cells to the BM. PMID- 11918089 TI - Daily mortality and fine and ultrafine particles in Erfurt, Germany part I: role of particle number and particle mass. AB - Increases in morbidity and mortality have been observed consistently and coherently in association with ambient air pollution. A number of studies on short-term effects have identified ambient particles as a major pollutant in urban air. This study, conducted in Erfurt, Germany, investigated the association of mortality not only with ambient particles but also with gaseous pollutants and indicators of sources. Part I of this study concentrates on particles. Data were collected prospectively over a 3.5-year period from September 1995 to December 1998. Death certificates were obtained from the local authorities and aggregated to daily time series of total counts and counts for subgroups. In addition to standard data for particle mass with diameters < or = 2.5 microm (PM2.5)* or < or = 10 microm (PM10) from impactors, a mobile aerosol spectrometer (MAS) was used to obtain size-specific number and mass concentration data in six size classes between 0.01 microm and 2.5 microm. Particles smaller than 0.1 microm were labeled ultrafine particles (three size classes), and particles between 0.1 and 2.5 microm were termed fine particles (three size classes). Concentrations of the gases sulfur dioxide (SO2), nitrogen dioxide (NO2), and carbon monoxide (CO) were also measured. The daily average total number concentration was 18,000 particles/cm3 with 88% of particles below 0.1 pm and 58% below 0.03 microm in diameter. The average mass concentration (PM2.5) was 26 microg/m3; of this, 75% of particles were between 0.1 and 0.5 microm in diameter. Other average concentrations were 38 microg/m3 for PM10, 17 microg/m3 for SO2, 36 microg/m3 for NO2, and 600 microg/m3 for CO. Ambient air pollution demonstrated a strong seasonality with maximum concentrations in winter. Across the study period, fine particle mass decreased, whereas ultrafine particle number was unchanged. The proportion of ultrafine particles below 0.03 microm diameter increased compared with the proportion of other particles. During the study, concentrations of SO2 and CO also decreased, whereas the concentration of NO2 remained unchanged. The data were analyzed using Poisson regression techniques with generalized additive modeling (GAM) to allow nonparametric adjustment for the confounders. Both the best single-day lag and the overall association of multiple days fitted by a polynomial distributed lag model were used to assess the lag structure between air pollution and death. Mortality increased in association with level of ambient air pollution after adjustment for season, influenza epidemics, day of week, and weather. In the sensitivity analyses, the results proved stable against changes of the confounder model. We saw comparable associations for ultrafine and fine particles in a distributed lag model where the contribution of the previous 4 to 5 days was considered. Furthermore, the data suggest a somewhat more delayed association of ultrafine particles than of fine particles if single-day lags are considered. The associations tended to be stronger in winter than in summer and at ages below 70 years compared to ages above 70 years. Analysis of the prevalent diseases mentioned on death certificates revealed that the overall association for respiratory diseases was slightly stronger than for cardiovascular diseases. In two-pollutant models, associations of ultrafine and fine particles seemed to be largely independent of each other, and the risk was enhanced if both were considered at the same time. Furthermore, when the associations were summed for the six size classes between 0.01 and 2.5 microm, the overall association was clearly stronger than the associations of the individual size classes alone. Associations were observed for SO2, NO2, and CO with mortality despite low concentrations of these gases. These associations disappeared in two-pollutant models for NO2 and CO, but they remained stable for SO2. The persistence of the SO2 effect was interpreted as artifact, however, because the SO2 concentration was much below levels at which effects are usually expected. Furthermore, the results for SO2 were inconsistent with those from earlier studies conducted in Erfurt. We conclude that both fine particles (represented by particle mass) and ultrafine particles (represented by particle number) showed independent effects on mortality at ambient concentrations. Comparable associations for gaseous pollutants were interpreted as artifacts of collinearity with particles from the same sources. PMID- 11918088 TI - The expression of rho proteins decreases with human brain tumor progression: potential tumor markers. AB - Astrocytic tumors are the most common human brain tumors. Establishment of tumor grade is a key determinant both in the choice of a therapeutic approach and in the prognosis. The diagnosis of astrocytic tumors is currently determined following histopathological analysis. The identification of molecular markers would offer a complementary tool for characterizing tumors with respect to their clinical behavior. In this study we determined the expression levels of 3 small GTP binding proteins (RhoA, RhoB and Rac1), of their inhibitor RhoGDI and of caveolin-1 in 24 human astrocytic tumors of grades I to IV. Our results demonstrated that the expression of RhoA and RhoB decreased significantly in all brain tumors studied and was inversely related with tumor of grade II to IV malignancy. The amount of caveolin-1 immunodetected was not significantly different from normal brain samples while the Rac1 expression level was diminished in astrocytic tumors of grades III and IV. Our finding that RhoA and RhoB expression levels are correlated to tumor malignancy suggests that they may serve as novel and efficient diagnostic markers for astrocytic brain tumors of histological grade II to IV and complement currently applied histopathological analysis. PMID- 11918090 TI - Childhood obsessive-compulsive disorder in the NIMH MECA study: parent versus child identification of cases. Methods for the Epidemiology of Child and Adolescent Mental Disorders. AB - Abstract-Because as many as 50% of obsessive-compulsive disorder (OCD) cases have had onset by age 15, interest in its detection in childhood is strong. Clinical experience indicates that children often try to keep their OCD secret and that parental report may give marked underestimates. The authors examined the prevalence of childhood OCD in the NIMH Methods for the Epidemiology of Child and Adolescent Mental Disorders (MECA) Study, a four-site community survey which allowed comparison of both parent and child report of the child's OCD and related symptoms and disorders. OCD cases, based on structured interviews (DISC-2.3 with DSM-III-R criteria) with 1,285 caretaker-child pairs, were identified separately for parent and child (aged 9 through 17) informants from the MECA database. Cases were then examined for demographic characteristics, for obsessive-compulsive symptoms and other diagnoses reported in cases "missed" by one reporter, and for comorbid disorders. Of a total of 35 (2.7%) identified cases, four (0.3%) were identified by the parent and 32 (2.5%) were identified by the child, with only one overlapping case. In general, when OCD cases were "missed" by one reporter, that reporter did not substitute another disorder. These findings support clinical data that children with OCD often hide their illness and underscore the importance of child interviews for its detection. PMID- 11918091 TI - The heritability of common phobic fear: a twin study of a clinical sample. AB - The aim of this study was to investigate the genetic and environmental contribution to common phobic fears, and to relate the findings to contemporary theories about the etiology of common phobic fears. Self-reported common phobic fear was studied in a treatment sample of 23 monozygotic and 38 same-sex dizygotic twin pairs. Heritability of .47 was observed for common phobic fear of small animals and social fear, and a heritability of .30 in common agoraphobic fear. For common fear of nature phenomena and situational fear, the heritability was 0. The finding that common nature and situational fears were solely caused by environmental factors is in support of learning theory, whereas results for animal, social, and other common phobic fear are in support of an integrative theory of biological preparedness, learning history, and a cognitive style of fearful expectation. PMID- 11918092 TI - Selective attention for hyperventilatory sensations in panic disorder. AB - According to cognitive theories, panic patients are assumed to display selective attention for feared bodily sensations. To date there has only been indirect evidence for this based on performance on reaction time tasks such as the modified Stroop task and the dot probe detection task. In the present study a more direct attempt was made to investigate whether panic patients show selective attention for feared sensations, in particular hyperventilatory sensations. A group of patients with panic disorder and a group of normal controls were compared on their performance on a reaction time task in which tones of different length had to be discriminated. During the task subjects were asked either to hyperventilate, to overbreathe under isocapnic conditions, or to make a neutral movement. No evidence for selective attention for hyperventilatory sensations was found. The article also discusses whether lack of finding interference during hyperventilation might be due to methodological issues. PMID- 11918093 TI - Adaptation as a sensorial profile in trait anxiety: a study with virtual reality. AB - Twenty-four volunteers were recruited on the basis of their trait anxiety scores (low trait anxiety [LTA] and high trait anxiety [HTA]) as assessed by the State Trait Anxiety Inventory. Adaptation to conflicting visual-vestibular stimulation (VVS) was used to study integration of space-related multi-sensory information in trait anxiety. First, vestibular perception was assessed by rotating the blindfolded subjects about the vertical axis (horizontal plane rotations) on a remote-controlled mobile robot. The subjects were asked to indicate the perceived rotation by use of an angular pointer. Subjects were then immersed into the center of a visual virtual square room by means of a head-mounted display. They were asked to control the robot with a joystick in order to perform 90 degrees rotations in the virtual room. However, a gain of 0.5 was introduced between visual scene and robot rotation so that the subjects were submitted to a conflict situation in which the 90 degrees rotational visual input was concurrent with a 180 degrees vestibular input. After 45 min of training with the virtual reality display, subjects were tested again in total darkness in order to determine whether their vestibular system had been reset by the conflicting visual signals. We found significant differences in adaptation to VVS between HTA and LTA groups as well as between males and females. Subjects of the HTA group demonstrated larger adaptation than that of the LTA group. Males also showed a greater level of adaptation compared to females. Our results suggest greater visual dependence in HTA subjects. This might be important for understanding the mechanisms underlying pathological anxiety and particularly agoraphobia. PMID- 11918094 TI - Overlapping and distinctive features of hypochondriasis and obsessive-compulsive disorder. AB - The spectrum of obsessive-compulsive disorders has received a great deal of theoretical attention, but there has been relatively little associated empirical research. The purpose of this study was to compare three groups of patients; those diagnosed with hypochondriasis (HC, a proposed spectrum condition), obsessive-compulsive disorder (OCD) and those with both OCD and HC (OCD/HC). The results show that patients with HC scored highest on a measure of overvalued ideas, and that the HC and HC/OCD groups scored significantly higher on measures of panic and agoraphobic cognitions. The groups also differed significantly for symptoms associated with compulsions. The patient groups were not different for measures of obsessions, depression, and anxiety. The results provide partial support for inclusion of HC in the spectrum of obsessive-compulsive disorders, but also provide indirect support for the association between HC and panic disorder. These results are interpreted in light of distinguishing characteristics among obsessive-compulsive spectrum conditions. PMID- 11918095 TI - The impact of anxiety sensitivity, bodily expectations, and cultural beliefs on menstrual symptom reporting: a test of the menstrual reactivity hypothesis. AB - According to the menstrual reactivity hypothesis, certain women report more severe as well as a greater number of menstrual symptoms due to accurate reports of physical symptoms and expectations (e.g., cultural beliefs, sex roles, bodily sensations). To test this hypothesis and to further examine the role that anxiety sensitivity plays in menstrual symptom reporting, women varying in levels of anxiety sensitivity completed measures of sex role socialization, menstrual attitudes, bodily preoccupations, affect, and fear of illness. To assess psychophysiological reactivity and the applicability of response styles theory to individuals varying in anxiety sensitivity, skin conductance was measured as participants engaged in a rumination or distraction task. In addition, participants completed a modified Stroop task consisting of anxiety, menstrual, and neutral words followed by a surprise recognition task. Retrospectively and prospectively, women high in anxiety sensitivity consistently reported more severe menstrual symptoms. High anxiety sensitivity women also reported preoccupation with bodily sensations and more negative attitudes toward illness, but did not differ from low anxiety sensitivity women on measures of menstrual attitudes or sex role socialization. While engaging in a rumination task, high anxiety sensitivity women exhibited more frequent skin conductance responses and greater skin conductance response magnitude than low anxiety sensitivity women. In addition, after the rumination task, high anxiety sensitivity women recognized more anxiety-related words from the Stroop task. Menstrual cycle phase had less of an impact than anxiety sensitivity level on the current results. Implications of these results for the menstrual reactivity hypothesis and rumination are discussed. PMID- 11918096 TI - Theory innocent or theory smart? PMID- 11918097 TI - The social construction of disability in schizophrenia. AB - Chronicity and disability can accompany a diagnosis of schizophrenia, but long term follow-up studies are demonstrating that these outcomes are not inevitable. There is growing awareness that characteristics of long-term illness previously understood to be part of the disorder can be partially constructed through a convergence of physical, psychological, and social processes. In this study, the potential for social construction of disability was explored through secondary analysis of qualitative data. Findings suggest that family, consumers, professionals, and society each have a role in shaping a person with schizophrenia's perceptions of his or her worth, competence, and place in society but that the individuals are not passive recipients of this input. PMID- 11918098 TI - Reconciling messages: the process of sexual talk for Latinas. AB - Young Latinas' experiences and meanings of sexuality were explored through 31 interviews, with grounded theory used to provide theoretical understanding of sexual talk. The women's level of comfort with sexual talk and their explicitness influenced the kind of sexual talk they engaged in with partners. The analysis of being in a romantic relationship revealed similar descriptions of processes within the accounts that led to the discovery of the core variable reconciling messages. This describes the Latinas' process of accepting messages that fit their value system, rejecting messages that theyfeel misrepresent their beliefs, and altering messages to accept their own sexuality. The findings expand Communication Boundary Management theory and have implications for interventions aimed at assisting women with safer sex topics. PMID- 11918099 TI - "You can't cure it so you have to endure it": the experience of adaptation to diabetic renal disease. AB - In this study, the experience of adaptation to diabetic renal disease was examined from a phenomenological perspective. Twenty patients attending a diabetic renal clinic were interviewed in depth. Through the use of a template analysis approach, a set of strong themes relating to changes in lifestyle was identified: changes in the nature of involvement with the medical system, coping strategies, and hopes, fears, and expectations. Almost all participants attempted to construct a "good adaptation" in the face of the uncertainties surrounding their condition by adopting a stoic and fatalistic stance. This is discussed in the context of the claim that contemporary society holds emotional self expression rather than stoical endurance to be the appropriate response to suffering. PMID- 11918100 TI - What is this thing called hormone replacement therapy? Discursive construction of medication in situated practice. AB - The use of hormone replacement therapy (HRT) involves complex decisions for mid aged women owing to controversy about the meaning of menopause and uncertainty regarding risks and benefits. Qualitative studies show that women can hold apparently contradictory beliefs, for example, both resisting and relying on medicalization. Focus group data (48 participants) and discourse analysis theorizing were used to investigate the complex discursive field available to women to construct HRTand to explain apparent contradictions. Interpretative repertoires identified in this study (threatening change, natural, biomedical, and drug) support previous findings. It is not contradictory to use different repertoires to achieve different discursive acts. The application of these findings to the development of decision tools that help women to arrive at individually appropriate decisions is discussed. PMID- 11918101 TI - Redefining life while forestalling death: living with an implantable cardioverter defibrillator after a sudden cardiac death experience. AB - Secondary analysis is a resourceful approach for making further use of existing data sets to answer questions not previously addressed or to expand on content not specifically examined in the original study. Using Heideggerian hermeneutics, the author analyzed transcripts of the stories of individuals with implantable cardioverter defibrillators (ICD) from three of her earlier phenomenological studies to obtain a description of the experience of living with an ICD after a sudden cardiac death experience. Three related themes emerged: (a) losing control: technology as lifesaving yet changing everything; (b) getting on with living: regaining control or conditional acceptance; and (c) creating a new vision: transformation or tenuous truce. The constitutive pattern was redefining life while forestalling death. PMID- 11918102 TI - Making the connection: seeking and receiving help by elderly African Americans. AB - The process of help seeking and help giving by older rural African Americans was studied using interviews, conversations, and participant observation of 41 informants, including 9 key informants. The author used an inductive analytical approach to examine the data, and a culturally constructed pattern of behavior was identified as help seeking. The circumstances under which, and language with which, help is sought and received was explained, and insight as to who is likely to receive help was provided. The elders believed that receiving help in late life is a reward for a good life. By understanding and following the rules of help seeking, formal helpers might more efficiently and effectively meet the needs of this vulnerable group. PMID- 11918103 TI - Making meaning: the creative component in qualitative research. AB - Findings in qualitative research are often wondrous and exciting, expounding new knowledge and perceptions previously unknown. Qualitative research requires the researcher to ponder and reflect on the data collected so as to find the meaning within. Helping researchers learn how to perform this step is not well discussed in the qualitative literature, yet this is one of the more crucial components of this type of research. In this article, the incubation, the meaning-making phase of qualitative research, is discussed in relation to the experiences of five researchers who have used traditional processes, models, metaphors, plays, pastiche, poetry, and quilt making and design to help them make meaning. PMID- 11918104 TI - Critical issues in interviewing people with traumatic brain injury. AB - People with cognitive impairments often cannot effectively recall and articulate experiences, feelings, and perceptions. Therefore, interviewing them can be fraught with pragmatic and methodological difficulties. Given this situation, the authors' experience in a study on the allocation of rehabilitative services to survivors of traumatic brain injury is discussed. Participants had physiological impairments affecting their ability to focus on interview questions. The authors discuss the unique challenges, including participants' recall of events, intolerance to stimuli, and image management and offer strategies for mitigating these concerns. The traditional view that reality is captured only in the subjective accounts of articulate people who can remember events in a precise and reflective manner is challenged. PMID- 11918105 TI - Talk that talk: storytelling and analysis rooted in African American oral tradition. AB - Stories are the foundation of qualitative research. However, the development of qualitative methods rooted in oral traditions remains largely unexplored by researchers. The contextual and historical influences on storytelling and storytaking are critical features of the African American oral tradition that are often ignored or minimized in qualitative research. Despite the complex and often contentious history of African Americans, their oral traditions have not been explored to reveal the depth of their lived experiences and the way those experiences inform their health concerns. The purpose of this article is twofold. First, dialogues about storytelling and storytaking are revisited and critiqued. Second, a comprehensive analytic process for gathering and interpreting stories rooted in African American oral tradition is outlined. PMID- 11918106 TI - Iron deficiency in autism and Asperger syndrome. AB - This research considers the prevalence of iron deficiency in children with autism and Asperger syndrome and examines whether this will influence guidelines and treatment. Retrospective analysis of the full blood count and, as far as available, serum ferritin measurements of 96 children (52 with autism and 44 with Asperger syndrome) was undertaken. Six of the autistic group were shown to have iron deficiency anaemia and, of the 23 autistic children who had serum ferritin measured, 12 were iron deficient. Only two of the Asperger group had iron deficiency anaemia and, of the 22 children who had their serum ferritin measured, only three were iron deficient. Iron deficiency, with or without anaemia, can impair cognition and affect and is associated with developmental slowing in infants and mood changes and poor concentration in children. This study showed a very high prevalence of iron deficiency in children with autism, which could potentially compromise further their communication and behavioural impairments. PMID- 11918107 TI - Harrdiness and social support as predictors of stress in mothers of typical children, children with autism, and children with mental retardation. AB - This study assessed the effects of social support and hardiness on the level of stress in mothers of typical children and children with developmental disabilities. One hundred and twenty mothers participated (40 mothers of children with autism, 40 mothers of children with mental retardation, and 40 mothers of typically developing children). Results indicated significant group differences in ratings of depression, anxiety, somatic complaints and burnout. Regression analyses were conducted to determine the best predictors of the dependent measures. Both hardiness and social support were predictive of successful adaptation. The relationships among hardiness, support and coping are discussed. PMID- 11918108 TI - The Scottish Centre for autism preschool treatment programme. II: The results of a controlled treatment outcome study. AB - This article evaluates the effectiveness of a developmentally based early intervention programme. Two groups of children were compared, a treatment group and a no-treatment control group. Standardized assessments were administered before and after the intervention period by an independent clinician. Pre treatment comparisons revealed that the control group had a significantly higher pre-treatment IQ; but the two groups were comparable for age, mental age, socioeconomic status and number of hours of non-experimental therapy. Results demonstrated that children in the treatment group improved significantly more than those in the control group on measures of joint attention, social interaction, imitation, daily living skills, motor skills and an adaptive behaviour composite. A measure of requesting behaviour fell short of statistical significance. The total stress index reduced for treatment group parents and increased for the control group parents (but not significantly). The results of the study are considered to support the efficacy of this treatment approach. PMID- 11918109 TI - Developing a diagnostic and intervention package for 2- to 3-year-olds with autism: outcomes of the frameworks for communication approach. AB - The aim of the research was to develop and evaluate a model of good practice which would make an explicit link between diagnosis and intervention, and so give parents a very clear rationale for the autism-specific yet individualized programme that they were carrying out. It employed an action research design, which essentially is responsive to participants, thus developing a user-friendly model of service. The programme was based on the developmental perspective that the pragmatics of language are the precursors of speech itself and enable both communication and relationship between child and parents. Since these are impaired in autism they should therefore be prioritized in early intervention. Ten children aged 1:10 to 2:9 at assessment, and with a diagnosis of autism, underwent an intervention based on home visits, modelling, workshops and written information, with parents as 'therapists' in naturally occurring situations. Within 18 months all children made substantial progress in social interaction and expressive communication, including gestural and verbal communication. PMID- 11918110 TI - Do children with autism learn to read more readily by computer assisted instruction or traditional book methods? A pilot study. AB - The study evaluates the progress of eight children aged 3-5 years with autism attending a specialist teaching unit in their development of reading skills in two conditions: computer instructed learning and book based learning. The authors developed a direct observation schedule to monitor autistic behaviours using computerized techniques. The children were matched by age, severity of autistic symptomatology and number of spoken words. They were initially randomly allocated to the computer or book condition and crossed over at 10 weeks. All of the children spent more time on task in the computer condition than in the book condition. By the end of the study after computer assisted learning, five of the eight children could reliably identify at least three words. It was found that children with autism spent more time on reading material when they accessed it through a computer and were less resistant to its use. PMID- 11918111 TI - The CAST (Childhood Asperger Syndrome Test): preliminary development of a UK screen for mainstream primary-school-age children. AB - The article describes a pilot and follow-up study of the preliminary development of a new tool to screen for Asperger syndrome (AS) and related social and communication conditions (the Childhood Asperger Syndrome Test, CAST) in children aged 4-11 years, in a non-clinical setting. In the pilot study, parents of 13 children with AS and of 37 typically developing children completed the CAST. There were significant differences between the AS and typical sample means. The pilot was used to establish preliminary cut-off scores for the CAST. In the main study, parents of 1150 primary-school-age children were sent the CAST, and 174 took part in the full data analysis. Results suggest that compared with other tools currently available, the CAST may be useful for identifying children at risk for AS and related conditions, in a mainstream non-clinical sample. Further research is ongoing. PMID- 11918112 TI - Human beings, animals and inanimate objects: what do people with autism like? AB - An experimental strategy based on the 'sorting by preference' approach was used to obtain information about the nature of the autistic syndrome. Twelve participants with autism (mean age 11:9 years), 12 with Down's syndrome (mean age 11:5 years) and 12 typically developing children (mean age 6:2 years) were matched on gender (M:F 9:3) and on verbal mental age. In a forced choice procedure they had to choose between: human beings or inanimate objects (relatedness condition); animals or inanimate objects (animate condition); drawings of a child handling a thing or of the same child in contact with another person (interpersonal relationship condition); pleasant or unpleasant situations without living beings (control condition). The performances of the groups differed only on the relatedness condition and on the interpersonal relationship condition. The results are discussed in the context of the social difficulties experienced by individuals with autism. PMID- 11918113 TI - Full-time equivalent (FTE) numbers. PMID- 11918114 TI - Promotion of hand hygiene: magic, hype, or scientific challenge? PMID- 11918115 TI - Limited impact of sustained simple feedback based on soap and paper towel consumption on the frequency of hand washing in an adult intensive care unit. AB - OBJECTIVE: To determine whether hand washing would increase with sustained feedback based on measurements of soap and paper towel consumption. DESIGN: Prospective trial with a nonequivalent control group. SETTING: Open multibed rooms in the Omaha Veterans Affairs Medical Center's Surgical Intensive Care Unit (SICU) and Medical Intensive Care Unit (MICU). SUBJECTS: Unit staff. INTERVENTION: Every weekday from May 26 through December 8, 1998, we recorded daytime soap and paper towel consumption, nurse staffing, and occupied beds in the SICU (intervention unit) and the MICU (control unit) and used these data to calculate estimated hand washing episodes (EHWEs), EHWEs per occupied bed per hour, and patient-to-nurse ratios. In addition, from May 26 through June 26 (baseline period) and from November 2 through December 8 (follow-up period), live observers stationed daily for random 4-hour intervals in the MICU and the SICU counted actual hand washing episodes (CHWEs). The intervention consisted of posting in the SICU, but not in the MICU, a graph showing the weekly EHWEs per occupied bed per hour for the preceding 5 weeks. RESULTS: Directly counted hand washing fell in the SICU from a baseline of 2.68+/-0.72 (mean +/- standard deviation) episodes per occupied bed per hour to 1.92+/-1.35 in the follow-up period. In the MICU, episodes fell from 2.58+/-0.95 (baseline) to 1.74+/-0.69. In the MICU, the withdrawal of live observers was associated with a decrease in estimated episodes from 1.36+/-0.49 at baseline to 1.01+/-0.36, with a return to 1.16+/-0.50 when the observers returned. In the SICU, a similar decrease did not persist throughout a period of feedback. Estimated hand washing correlated negatively with the patient-to-nurse ratio (r = -0.35 for the MICU, r = -0.46 for the SICU). CONCLUSIONS: Sustained feedback on hand washing failed to produce a sustained improvement. Live observers were associated with increased hand washing, even when they did not offer feedback. Hand washing decreased when the patient-to-nurse ratio increased. PMID- 11918117 TI - The role of nurse understaffing in nosocomial viral gastrointestinal infections on a general pediatrics ward. AB - OBJECTIVE: To examine the relationship between nurse staffing levels and the rate of nosocomial viral gastrointestinal infections (NVGIs) in a general pediatrics population. DESIGN: Retrospective descriptive study. SETTING: A general pediatrics ward at The Hospital for Sick Children in Toronto, Ontario, Canada, a 320-bed, tertiary-care pediatric institution. RESULTS: Forty-three NVGIs were detected in 37 patients of 2,929 admissions (1.3%). The monthly NVGI rate correlated significantly with the monthly night patient-to-nurse ratio (r = 0.56) and the monthly day patient-to-nurse ratio (r = 0.50). The nursing hours per patient-day during the preinfection period (PIP) were significantly lower than those during the nonpreinfection period (NPIP; 12.5 vs 13.0). There was no difference between the PIP and the NPIP day patient-to-nurse ratios (3.31 vs 3.32), but there was a significant difference between the PIP and the NPIP night patient-to-nurse ratios (3.26 vs 3.16). The incidence of NVGIs in the 72-hour period after any day when the nursing hours per patient-day were less than 10.5 was 6.39 infections per 1,000 patient-days, compared with 2.17 infections per 1,000 patient-days in periods with more than 10.5 nursing hours per patient-day (rate ratio, 2.94; 95% confidence interval, 2.16 to 4.01). CONCLUSION: Nurse understaffing contributed to an increased NVGI rate in our general pediatrics population, and should be assessed as a risk factor in outbreak investigations. PMID- 11918116 TI - How many nosocomial infections are associated with cross-transmission? A prospective cohort study in a surgical intensive care unit. AB - OBJECTIVE: To determine the percentage of cross-transmissions in an intensive care unit (ICU) with high nosocomial infection (NI) rates according to the data of the German Nosocomial Infection Surveillance System. SETTING: A 14-bed surgical ICU of a 1,300-bed, tertiary-care teaching hospital. METHOD: Prospective surveillance of NIs during a period of 9 months. If an NI was present, the isolates of the following indicator pathogens were stored and typed by species: Staphylococcus aureus, Enterococcus species, Escherichia coli, Pseudomonas aeruginosa, Acinetobacter baumannii, and Enterobacter species. Pulsed-field gel electrophoresis was performed for typing of S. aureus strains and arbitrarily primed polymerase chain reaction was applied for the other pathogens. The presence of two indistinguishable strains in two patients was considered as one episode of cross-transmission. RESULTS: Two hundred sixty-two patients were observed during a period of 2,444 patient-days; 96 NIs were identified in 59 patients and the overall incidence density of NI was 39.3 per 1,000 patient-days. For 104 isolates, it was possible to consider typing results. Altogether, 36 cross-transmissions have lead to NIs in other patients. That means at least 37.5% of all NIs identified were due to cross-transmissions. CONCLUSION: Because of the method of this study, the percentage of NIs due to cross-transmission identified for this ICU is an "at least number." In reality, the number of cross transmissions, and thus the number of avoidable infections, may have been even higher. However, it is difficult to assess whether the percentage of NIs due to cross-transmission determined for this ICU may be the crucial explanation for the relatively high infection rate in comparison to other surgical ICUs. PMID- 11918118 TI - Morbidity, mortality, and healthcare burden of nosocomial Clostridium difficile associated diarrhea in Canadian hospitals. AB - OBJECTIVE: To assess the healthcare burden, morbidity, and mortality of nosocomial Clostridium difficile-associated diarrhea (N-CDAD) in Canadian hospitals. DESIGN: Laboratory-based prevalence study. SETTING: Nineteen acute care Canadian hospitals belonging to the Canadian Hospital Epidemiology Committee surveillance program. PATIENTS: Hospitalized patients in the participating centers. METHODS: Laboratory-based surveillance was conducted for C. difficile toxin in stool among 19 Canadian hospitals from January to April 1997, for 6 continuous weeks or until 200 consecutive diarrhea stool samples had been tested at each site. Patients with N-CDAD had to fulfill the case definition. Data collected for each case included patient demographics, length of stay, extent of diarrhea, complications of CDAD, CDAD-related medical interventions, patient outcome, and details of death. RESULTS: We found that 371 (18%) of 2,062 tested patients had stools with positive results for C difficile toxin, of whom 269 (13%) met the case definition for nosocomial CDAD. Of these, 250 patients (93%) had CDAD during their hospitalization, and 19 (7%) were readmitted because of CDAD (average readmission stay, 13.6 days). Forty-one patients (15.2%) died, of whom 4 (1.5% of the total) were considered to have died directly or indirectly of N-CDAD. The following N-CDAD-related morbidity was noted: dehydration, 3%; hypokalemia, 2%; gastrointestinal hemorrhage requiring transfusion, 1%; bowel perforation, 0.4%; and secondary sepsis, 0.4%. The cost of N-CDAD readmissions alone was estimated to be a minimum of $128,200 (Canadian dollars) per year per facility. CONCLUSION: N-CDAD is a common and serious nosocomial infectious complication in Canada, is associated with substantial morbidity and mortality, and imposes an important financial burden on healthcare institutions. PMID- 11918119 TI - Preventing nosocomial transmission of pulmonary tuberculosis: when may isolation be discontinued for patients with suspected tuberculosis? AB - OBJECTIVE: The Centers for Disease Control and Prevention and the American Thoracic Society recommend obtaining cultures of at least three sputum specimens for acid-fast bacilli (AFB) from patients in whom tuberculosis (TB) is suspected. On the basis of this, most hospitals isolate patients with suspected TB for 3 days or more until three smear (not culture) results are negative. Our objective was to evaluate the predictive value and sensitivity of these smears. DESIGN: Observational study. SETTING: Four urban medical centers. METHODS: The posttest probability of TB given sequential negative AFB smears from 274 patients isolated for suspected TB and the sensitivity of sequential AFB smears from 209 patients with positive results on culture for pulmonary TB were measured. RESULTS: The posttest probabilities of having TB given one, two, and three negative AFB smears were low: 1.1% (3 of 265; 95% confidence interval [CI95], 0.23% to 3.27%), 0.4% (1 of 262; CI95 0% to 2.1%), and 0% (0 of 260; CI95, 0% to 1.4%), respectively. Among the 209 patients with positive results on culture for pulmonary TB, 169 (81%) had an expectorated sputum specimen sent, of which 91 (54%) were positive for AFB. Forty (24%) of the 169 patients had a second expectorated sputum specimen sent after the results of the first specimen were negative; only 6 (15%) of these had positive AFB smears. None of the 10 patients in whom the first two expectorated sputum samples yielded an AFB smear without an organism had a third AFB smear that was positive. CONCLUSION: Unless there is high clinical suspicion of pulmonary TB in a specific patient, the use of three AFB smears on expectorated sputa is a rational approach to discontinuing isolation for patients with suspected TB. PMID- 11918120 TI - The use of heteroduplex analysis of polymerase chain reaction products to support the possible transmission of Legionella pneumophila from a malfunctioning automobile air conditioner. AB - Air conditioner condensates have not been previously associated with cases of Legionnaires' disease. We report the possible transmission of Legionella pneumophila serogroup 1 from a malfunctioning automobile air conditioning system's leaking water onto the floorboard of a car driven for a long distance by the patient. Heteroduplex analysis of polymerase chain reaction products was used to help establish an epidemiologic link between the water specimen and the patient. PMID- 11918121 TI - Scissors: a potential source of nosocomial infection. AB - This point-prevalence survey of healthcare workers' scissors demonstrates that of 232 scissors sampled, 182 (78.4%) were colonized with bacteria. The scissors of nurses and those for communal use were most frequently contaminated. Cleaning of scissors occurred infrequently, but wiping scissors with an alcohol swab effectively disinfected them. The suspicion that scissors may harbor and potentially transmit microorganisms, including antibiotic-resistant bacteria, has been confirmed. PMID- 11918123 TI - A nursery outbreak of Staphylococcus aureus pyoderma originating from a nurse with paronychia. AB - An outbreak of methicillin-susceptible Staphylococcus aureus pyoderma occurred in the nursery of a tertiary-care referral center. All strains retrieved from the outbreak, including one from a nurse's infected finger, were typed by arbitrarily primed polymerase chain reaction and automated ribotyping. The results indicated that the spread of the outbreak was probably facilitated by contamination of the nurse with paronychia. PMID- 11918122 TI - Characteristics of a large cluster of vancomycin-resistant enterococci in an Australian hospital. AB - Investigation of a cluster of hospital inpatients colonized with vancomycin resistant enterococci (VRE) revealed a point-prevalence of 19.1%. The outbreak was controlled using established guidelines and additional strategies, including minimizing patient transfers and enforcing appropriate antibiotic prescribing. The incidence of VRE has remained low during the ensuing 30 months. PMID- 11918124 TI - Antimicrobial use prior to the acquisition of multiresistant bacteria. AB - We assessed whether patients who acquired methicillin-resistant Staphylococcus aureus (MRSA) had less exposure to antimicrobial agents than did those who acquired Enterobacteriaceae that produced extended-spectrum beta-lactamase (ESbetaL). In a 6-month, prospective study, ESbetaL carriers had received antimicrobial therapy more often than had MRSA carriers. Amoxicillin-clavulanic acid, fluoroquinolones, and third-generation cephalosporins, especially ceftazidime, had been prescribed more often for ESbetaL carriers than for MRSA carriers. PMID- 11918125 TI - Pneumonia in the long-term-care facility. AB - Pneumonia is a common infection among residents of long-term-care facilities (LTCFs), with an incidence of 1.2 episodes per 1,000 patient-days. This rate is believed to be six- to tenfold higher than the rate of pneumonia among elderly individuals living in the community. The risk factors for pneumonia among residents of LTCFs are profound disability, bedridden state, urinary incontinence, difficulty swallowing, malnutrition, tube feedings, contractures, and use of benzodiazepines and anticholinergic medications. An elevated respiratory rate is often an early clue to pneumonia in this group of patients. Staphylococcus aureus (including methicillin-resistant S. aureus) and aerobic gram-negative bacilli (including multidrug-resistant isolates) are more frequent causes of pneumonia in this setting than in the community. Criteria have been developed that help identify patients for treatment in their LTCFs. PMID- 11918126 TI - Chronic kidney disease: stating the managed care case for early treatment. Discussion and consensus of presentations of economic analyses, managed care organization case studies, and opportunities for intervention in a managed care setting. May 3-5, 2001 Chicago, Illinois. AB - Based on an analysis of data from the National Health and Nutrition Examination Survey, approximately 20 million people currently have some degree of decreased glomerular filtration rate or overt kidney disease. Despite numerous challenges to be overcome, managed care organizations and the nephrology community are seeking ways to collaborate in improving the quality of care of such patients. Using current data, as well as information about the cost of care, priority is being given to the establishment of early intervention programs to maximize the existing healthcare infrastructure and to improve the clinical outcomes of chronic kidney disease and end-stage renal disease. PMID- 11918127 TI - Evolution of all-cause mortality rates in the United States of America, anno 1900 1999. PMID- 11918129 TI - Effect of phosphodiesterase III inhibitor on plasma concentrations of endothelin 1 and tumour necrosis factor in patients with acute heart failure. AB - OBJECTIVE: Effects of loprinone hydrochloride on plasma ET-1 and TNF-alpha levels were assessed in patients with acute heart failure (AHF). METHODS AND RESULTS: Thirty patients with AHF were divided into 2 groups and treated with loprinone hydrochloride (0.3 pg/kg/min) (n = 15) or placebo (n = 15). Twenty healthy controls were also included. Plasma ET-1 and TNF-alpha were significantly higher in the 30 AHF patients than in the healthy controls. In AHF patients, loprinone hydrochloride lowered plasma ET-1 and TNF-alpha (p < 0.01). CONCLUSION: ET-1 and TNF-alpha may play pathophysiological roles in the progression of AHF. Loprinone hydrochloride is effective in reducing plasma ET-1 and TNF-alpha levels in AHF patients. PMID- 11918128 TI - Does exercise-induced severe ischaemia result in elevation of plasma troponin-T level in patients with chronic coronary artery disease? AB - OBJECTIVE: It has been reported that the loss of cell membrane integrity during severe reversible ischaemia results in elevation of plasma troponin T (TnT) in unstable angina. We investigated whether TnT is released into circulation during severe ischaemia (e.g. on treadmill exercise testing) in patients with chronic coronary artery disease (CAD). METHODS AND RESULTS: The study comprised 54 patients who had angiographically documented CAD (22 of them had a prior history of myocardial infarction) and 18 normal subjects. All cases underwent exercise thallium (Tl)-201 SPECT myocardial perfusion imaging. Blood samples for TnT analysis were obtained for each patient at pre-exercise, immediately after and 12 hours after exercise. SPECT images were divided into 20 segments. Patients with redistribution defects > or = 5 were considered to have severe ischaemia. Sixteen patients had severe ischaemia on SPECT images. The mean TnT levels of the patients with severe ischaemia at pre-exercise, immediately after and 12 hours after exercise were 0.009 +/- 0.008, 0.012 +/- 0.009 and 0.010 +/- 0.010 ng/ml, respectively. In normal subjects the mean TnT levels were 0.012 +/- 0.009, 0.010 +/- 0.009, 0.010 +/- 0.010 ng/ml, respectively. There was no significant difference neither between the two groups nor within each group with respect to pre-exercise and post-exercise TnT results. All TnT levels were within normal limits (< 0.1 ng/dl). Additionally, when the patients were grouped according to other SPECT variables (patients with reversible defects vs. fixed defects; patients with increased lung uptake of Tl-201 vs. without lung uptake; patients with transient left ventricle dilatation vs. those without) and angiographic findings (patients with multivessel disease vs. normal subjects), there were no significant differences with respect to pre-exercise and post-exercise TnT results. CONCLUSION: Severe ischaemia does not result in the elevation of the plasma TnT level in patients with chronic CAD. PMID- 11918130 TI - Ventricular tachycardia induced by a pacemaker lead. AB - The authors present a case of symptomatic non-sustained ventricular tachycardia induced by mechanical irritation of the outflow tract of the right ventricle by a loop in a permanent ventricular pacing lead. Reposition of the lead eliminated the rhythm disturbance. PMID- 11918131 TI - Papillary muscles identified by myocardial contrast echocardiography in preparation for percutaneous transluminal septal myocardial ablation. AB - In patients with hypertrophic obstructive cardiomyopathy (HOCM), percutaneous transluminal septal myocardial ablation (PTSMA) supplemented by myocardial contrast echocardiography (MCE) can provide a therapeutic benefit. We report a 33 year-old woman with HOCM in which MCE revealed a septal branch artery supplying the postero-medial papillary muscles. Thus, PTSMA was aborted because of the risk of acute mitral regurgitation secondary to papillary muscle dysfunction. PMID- 11918132 TI - Reversal of left ventricular hypertrophy with the ACE inhibitor moexipril in patients with essential hypertension. PMID- 11918133 TI - GPIIb/IIIa polymorphism in patients with myocardial infarction. PMID- 11918134 TI - Effect of blood pressure normalization on endothelial function evaluated by high resolution ultrasonography in patients with essential hypertension. PMID- 11918135 TI - Heart rate variability in patients after coronary artery bypass grafting: early and late effects of cardiac rehabilitation. PMID- 11918136 TI - Pulse wave velocity and the estimated risk of stroke and myocardial infarction. PMID- 11918137 TI - Echocardiography in patients with destructive aortic valve infective endocarditis. PMID- 11918138 TI - Transoesophageal Doppler echocardiography provided important parameters in patients with dilated cardiomyopathy during bisoprolol therapy. PMID- 11918139 TI - Relationship between carotid intima-media thickness, atherosclerosis risk factors and angiographic findings in patients with coronary artery disease. PMID- 11918140 TI - Baroreflex sensitivity: population values. PMID- 11918141 TI - Accurate accelerated 3-dimensional echocardiographic quantification of left ventricular function in unselected patients: a comparison with radionuclide angiography. PMID- 11918142 TI - Echocardiographic identification of resting perfusion defects using bolus administration of Levovist and power contrast imaging. PMID- 11918144 TI - Changes in regional myocardial velocities at an infarct zone before and after PTCA--a tissue Doppler imaging study. PMID- 11918143 TI - Preliminary experience with diagnostic application of resting intravenous myocardial contrast echocardiography in suspected myocardial infarction. PMID- 11918145 TI - Determination of left ventricular ejection fraction by gated 99mTc-sestamibi SPECT--correlation with coronary angiography. PMID- 11918146 TI - Coronary artery calcification and dietary cholesterol intake in Korean men. AB - OBJECTIVE: This study was performed to examine the relationship between dietary cholesterol intake and coronary artery calcification (CAC) score in healthy men. METHODS: Electron beam computed tomography (EBCT) was used to examine the CAC score in 135 Korean men aged 40-81 years who did not have clinical illness. Dietary cholesterol intake was assessed by a nutritionist using a semiquantitative food frequency method. Body mass index (BMI), serum lipid levels, cigarette use, alcohol intake, exercise, and a past history of cardiovascular disease were determined during interview and examination. RESULTS: The resultant median CAC score among those who experienced CAC was 22.5 (1-697) and average intakes of total fat and cholesterol were 22.4% (13.8-40.7) of total energy intake and 306.0 mg/day (84-1191). When the participants were classified into high (> or = 75 percentile) and low (< 75 percentile) CAC score groups, multiple logistic analysis showed that the cholesterol intake (per 10 mg/1000 kcal of energy) was significantly related to a high CAC score (OR 1.12: 95% CI 1.02-1.24), after adjustment for age, BMI, serum triglyceride level, past history of hypertension, past history of hyperlipidaemia, and energy intake. Also, when participants were classified into 2 groups (CAC score > or = 100 vs. < 100), cholesterol intake was found to be significantly related to CAC score. However, fatty acid intakes were not significantly related to the CAC score. CONCLUSION: These results suggest that in a population with a relatively low risk of coronary heart disease, higher cholesterol intake may increase the level of CAC. PMID- 11918147 TI - Seismocardiography--a non-invasive technique for estimating left ventricular function: preliminary results. PMID- 11918149 TI - Influence of programmed AV delay on left ventricular diastolic function in DDD paced hypertensive patients. PMID- 11918148 TI - Influence of left ventricular regional myocardial ischaemia on regional systolic and diastolic function in ischaemic heart disease patients. PMID- 11918150 TI - Quantitative assessment of mitral regurgitation: comparison of echocardiographic and angiographic methods. PMID- 11918151 TI - Dobutamine stress echocardiography in symptomatic patients with nondiagnostic or negative treadmill exercise test. PMID- 11918152 TI - Blood pressure variability in paced patients with sick sinus syndrome. PMID- 11918153 TI - Acoustic quantification assessment of left atrial area and function in patients with dilated nonischaemic cardiomyopathy with and without restriction in Doppler study. PMID- 11918154 TI - Role of contrast echocardiography in the improvement of left ventricular endocardial border visualization. PMID- 11918155 TI - Additional effects of irbesartan when compared to captopril in the prevention of post-infarction left ventricular dilatation. PMID- 11918156 TI - Assessment of stent patency using multi-slice spiral computed tomography: initial experience. PMID- 11918157 TI - Assessment of the patency of coronary artery bypass grafts using multi-slice spiral computed tomography: initial experience. PMID- 11918158 TI - QT interval dispersion in patients with insulin-dependent diabetes mellitus after acute myocardial infarction. PMID- 11918159 TI - Prognostic value of colour Doppler echocardiography in patients after myocardial infarction--2-year follow-up. PMID- 11918160 TI - Tissue Doppler echocardiography during low-dose dobutamine infusion in patients with ischaemic and idiopathic dilated cardiomyopathy. PMID- 11918161 TI - Evaluation of the effect of mitral prolapse on positive tilt test results in children with syncope of unknown origin. PMID- 11918162 TI - Heart rate variability in patients treated with percutaneous transluminal coronary angioplasty. PMID- 11918163 TI - Bradyarrhythmias, conduction disturbances and the need for permanent pacing after aortic valve replacement. PMID- 11918164 TI - Validation of two-dimensional echocardiography for quantifying left ventricular aneurysm: comparison with magnetic resonance imaging and evaluation during cardiac surgery. PMID- 11918165 TI - Comparative analysis of ambulatory self-measurement of blood pressure and automatic ambulatory blood pressure monitoring. PMID- 11918166 TI - Cardiopulmonary exercise capacity in adults with atrial septal defect. PMID- 11918167 TI - Resting ventilatory abnormalities in adults with atrial septal defect. PMID- 11918168 TI - Differences between right ventricular contractility in newborns with preload and afterload changes in selected congenital heart defects. PMID- 11918169 TI - Rapid resolution of ST-segment elevation predicts functional recovery in patients with acute myocardial infarction treated with primary percutaneous transluminal coronary angioplasty. PMID- 11918170 TI - Lung function and exercise tolerance in patients with heart failure. PMID- 11918171 TI - Prognostic assessment of patients with heart failure--based on the non-invasive procedures. PMID- 11918173 TI - Influence of low dobutamine dose on left ventricular diastolic flow parameters in subjects with low ejection fraction. PMID- 11918172 TI - Perfusion myocardial scintigraphy as a method of qualification and control for percutaneous transmyocardial laser revascularization in patients with end-stage coronary artery disease. PMID- 11918174 TI - Atrial cardiomyopathy as a consequence of atrial fibrillation. PMID- 11918175 TI - Congenital tumours and tumour-like lesions in domestic animals. 1. Cattle. A review. AB - The literature on congenital tumours and tumour-like lesions in calves was reviewed. Lesions were subdivided by their anatomical distribution and in addition also according to their histologic-pathogenetic nature. As a result of the latter method, four main groups were formed covering most of the lesions described so far: malignant lymphomas, mesotheliomas, hamartomas and embryonic tumours. Most lesions were of mesenchymal structure, carcinomas being extremely rare. Some findings may point to early genetic events, for instance twin calves both affected with malignant lymphoma and related calves with congenital facial neurofibromatosis. An external factor, asbestos, is suspected to play a role in the genesis of peritoneal mesotheliomatosis. The effects of congenital tumours on their hosts were often considerable: death by generalization (malignant lymphomas), ascites (mesotheliomas) or the growth of large abdominal tumours (nephroblastomas, mixed tumours). The latter two conditions often caused dystocia. PMID- 11918176 TI - Paratuberculosis with special reference to cattle. A review. AB - Paratuberculosis is a chronic, granulomatous enteritis caused by Mycobacterium avium subspecies paratuberculosis affecting domestic and wild ruminants. The symptoms of clinical paratuberculosis are chronic diarrhoea and progressive weight loss while subclinically infected animals mainly have decreased production. The infection is widespread throughout the world and causes substantial financial losses for the farming industry. One of the major obstacles in the control of this disease, is the difficulty of identifying subclinically infected animals. This review gives a summary of several aspects of paratuberculosis including clinical importance, pathology, immunology and properties of the infectious agent. Special emphasis will be on the available diagnostic methods, their use and limitations. PMID- 11918178 TI - Characterization of metal oxide surfaces and thin semiconductor films by inelastic electron tunneling spectroscopy. AB - Inelastic electron tunneling spectroscopy (IETS) is a unique surface and interface analytical technique using electron tunneling through a metal/insulator/metal tunneling junction at cryogenic temperatures. It gives the vibrational spectrum of a very thin (nm) insulator film and the adsorbed species on it. The high sensitivity, good resolution, and wide spectral range inherent in IETS enable us to analyze the surface and interface of the insulator in detail. The tunneling junction is a good model system for oxide catalysts, electronic devises, and solid state sensors. Information about the surfaces of alumina and magnesia, the adsorption states and chemical reactions of adsorbed species occurring on these oxides can be obtained through an analysis of the tunneling spectra. The structures and properties of evaporated thin semiconductor films can also be studied. In this review, the surface characterization of alumina and magnesia, the adsorption and surface reactions of organic acids, esters, amides, and nitryls on these oxides, and the characterization of thin evaporated films of Si, Ge, and the oxides are summarized. PMID- 11918177 TI - Clinical efficacy of florfenicol in the treatment of calf respiratory tract infections. AB - This paper reports on a study of the aetiology of calf pneumonia and the clinical efficacy of florfenicol, a new antibiotic in Turkey. Twenty-seven weaned and unweaned calves (13 males and 14 females) between 1 and 16 months of age brought to the clinics of Selcuk University, Faculty of Veterinary Science. Broncho alveolar lavage (BAL) fluid samples were taken from the animals diagnosed to have upper respiratory tract infection associated with bronchitis (N=2), bronchitis (N=5), bronchopneumonia (N=4), pneumonia (N=3), pleuropneumonia (N=11), bronchopneumonia plus pulmonary oedema (N=2) based on the results of the clinical and laboratory examinations. Then microbiological isolation and antibiotic culturing were performed. The animals were treated with 1 ml/15 kg (20 mg/kg) florfenicol (Nuflor, DIF) twice within 48 hours via intramuscular injection. At the end of the treatment, 23 of the weaned and unweaned calves were completely healed, 1 calf had died and 3 calves showed no healing. The results of BAL samples and microbiological examinations of the 3 calves that did not respond to the treatment indicated that these cases were affected by mixed infections of yeasts, fungi, and bacteria. Widespread pleuropneumonia was observed. According to the results of the microbiological examination of the BAL samples, Mannheimia (Pasteurella) haemolytica had the highest isolation rate (25%) compared with the other isolated bacteria, namely, Klebsiella pneumonia (20%), Actinomyces pyogenes (15%), beta-hemolytic streptococci. (10%), Staphylococcus spp. (5%), and E. coli (5%). The study also revealed fungi [Penicillum spp. (5%) and Aspergillus spp. (5%)] and two calves (10%) had a yeast infection.. We conclude that florfenicol has a high bacteriological and clinical efficacy (100% and 96% respectively) in the treatment of calf respiratory tract diseases. PMID- 11918179 TI - A tunable picosecond dye laser for use in dioxin analysis. AB - A distributed-feedback dye laser with a quenching cavity was designed and constructed for generating a tunable picosecond pulse with a narrow spectral linewidth. This nearly transform-limited pulse was succeedingly amplified by a triple-pass off-axis amplifier. The pulse duration and the spectral linewidth were 60 ps and 9.4 pm, respectively. The amplified pulse was frequency-doubled by second-harmonic generation, producing a 0.5-mJ pulse with no background emission. The potential advantage of this laser in the analysis of dioxin based on supersonic jet/resonance-enhanced multiphoton ionization/mass spectrometry is discussed. PMID- 11918180 TI - Achieving high time-resolution with a new flow-through type analyzer for total inorganic carbon in seawater. AB - A fully automated, continuous-flow-through type analyzer was developed to observe rapid changes in the concentration of total inorganic carbon (CT) in coastal zones. Seawater and an H3PO4 solution were fed into the analyzer's mixing coil by two high-precision valveless piston pumps. The CO2 was stripped from the seawater and moved into a carrier gas, using a newly developed continuous-flow-through CO2 extractor. A mass flow controller was used to assure a precise flow rate of the carrier gas. The CO2 concentration was then determined with a nondispersive infrared gas analyzer. This analyzer achieved a time-resolution of as good as 1 min. In field experiments on a shallow reef flat of Shiraho (Ishigaki Island, Southwest Japan), the analyzer detected short-term, yet extreme, variations in CT which manual sampling missed. Analytical values obtained by the analyzer on the boat were compared with those determined by potentiometric titration with a closed cell in a laboratory: CT(flow-through) = 0.980 x CT(titration) + 38.8 with r2 = 0.995 (n = 34; September 1998). PMID- 11918182 TI - Application of a surface plasmon resonance sensor to analyses of amine compounds with the use of a polymer film and an acid-base reaction. AB - A surface plasmon resonance (SPR) sensor was applied to analyses of some amine compounds (n-butylamine, isobutylamine, aniline, and N,N-dimethylaniline) by using a polymer film and an acid-base reaction in it. Poly(acrylamide) (PAA) was adopted as the polymer film and was immobilized on an Au film to prepare a sensor chip. Pivalic acid was entered into the PAA film as an acid. The PAA film with a thickness of 50 nm gave the highest sensitivity to the SPR sensor. Although water was better concerning the sensitivity for the SPR sensor as the solvent, ethanol was adopted because it dissolves well all of the amine compounds used. The Au film coated with the PAA film gave higher sensitivity for analyses of n butylamine and isobutylamine, and lower sensitivity for analyses of aniline and N,N-dimethylaniline than an Au film without the PAA film. The PAA film containing pivalic acid gave 4-5 orders of magnitude higher sensitivity to the SPR sensor for analyses of all the amine compounds due to the reaction between pivalic acid and these amine compounds. PMID- 11918181 TI - Determination of total mercury and methylmercury in human hair by graphite furnace atomic absorption spectrophotometry using 2,3-dimercaptopropane-1 sulfonate as a complexing agent. AB - For the determination of total mercury in hair, an amount (25.0 mg) of hair sample was digested with conc. HNO3 (400 microl) at 90 degrees C for 10 min in a 7-ml teflon microreaction vessel. After digestion, the pH of the acidic hair mixture was adjusted to 5.0-6.0 by NaOH and was then passed through a clean-up Sep-Pak C18 cartridge. To the eluate, 2,3-dimercaptopropane-1-sulfonate (DMPS) and sodium acetate buffer (pH = 6.0) were added to form a mercury-DMPS complex. This complex was preconcentrated on two Sep-Pak C18 cartridges in series, and each cartridge was eluted with methanol and adjusted to 2.00 ml. A portion (50 microl) was introduced into a graphite cuvette and then atomized according to a temperature program. The method detection limit (MDL, 3sigma) was 0.064 (microg g(-1)); the calibration graph was linear up to 7.52 microg g(-1). Good accuracies were obtained when testing two human hair certified reference materials (GBW 09101 and BCR-397). Six real samples were analyzed, and the recoveries were 95.8 98.2% with a relative standard deviation (RSD, n = 3) < 2.1%. For the determination of methylmercury (CH3Hg+), 25.0 mg of hair sample was extracted with 2.0 mol dm(-3) HCl (1.0 ml) by ultrasonicating for 1 h. The supernatant solution was used for CH3Hg+ analysis and the hair residue was used for the analysis of inorganic mercury (Hg2+). The MDL of CH3Hg+ was 0.068 microg g(-1); the calibration graph was linear up to 6.00 microg g(-1). Six real samples were analyzed, and the recoveries were 96.0-99.2% with RSD (n = 3) < 2.3%. The sum of the concentrations of CH3Hg+ and Hg2+ was very close to that of the total mercury measured with a relative error within 3.6%. The proposed method can be accurately applied to the measurement of CH3Hg+, Hg2+, and total mercury in hair samples. PMID- 11918183 TI - Characteristics of a surface plasmon resonance sensor combined with a poly(vinyl chloride) film-based ionophore technique for metal ion analyses. AB - The characteristics of a surface plasmon resonance (SPR) sensor prepared by coating a metal film evaporated on a prism with a polymer film containing tetra-n butyl thiuram disulfide (TBTDS) were studied. The differences in the sensitivity, selectivity, and detection limit for a Zn2+ ion of the SPR sensor were reported as a function of the thickness of the polymer film, the kind of a metal film, and the kind of a polymer film. The thinner was the polymer film, the higher was the sensitivity, and the lower was the detection limit. The Ag film gave to the SPR sensor higher sensitivity than the Au film. TBTDS contained in the poly(vinyl chloride) (PVC) film slightly improved the selectivity toward the Zn2+ ion. A non conditioned poly(methyl methacrylate) (PMMA) film containing TBTDS gave a lower detection limit of 1.0 x 10(-6) mol/l, which is similar to that obtained by using an ion selective electrode (ISE) method, than the PVC film. The PVC film, however, gave higher concentration resolution than the PMMA film. PMID- 11918184 TI - Uptake of transition metal ions using liposomes containing dicetylphosphate as a ligand. AB - The uptake of Cu2+ was investigated using various types of liposomes composed of phosphatidylcholine (PC), cholesterol (Chol) and dicethylphosphate (DCP). DCP played a role as a ligand for Cu2+. Multilamellar vesicles (MLVs) were more effective for the uptake of Cu2+ compared to unilamellar vesicles prepared by the extrusion technique. The uptake efficiency of MLVs for Cu2+ was dependent on the molar ratio of DCP in MLVs. The uptake percent of Cu2+ was 92% using MLVs having a PC:DCP:Chol molar ratio of 4:3:3; 95% of the total vesicle Cu2+ was bound to DCP of the outer membrane surface of the MLVs, and the remaining 5% of the total Cu2+ was distributed into the interior side of the MLVs. MLVs having a PC:DCP:Chol molar ratio of 4:3:3 were also effective as separation media for Mn2+, Co2+, Ni2+ and Zn2+. The uptake efficiency of the MLVs for the transition metal ions increased in the order Co2+ < Zn2+ < Ni2+ < Mn2+ < Cu2+. PMID- 11918185 TI - Characterization of oligomeric polypropyleneglycol acrylate by GC, SFC and MALDI TOF-MS. AB - Polypropyleneglycol acrylate (PGA), one of the typical acrylic oligomers manufactured industrially, was comprehensively characterized by gas chromatography (GC), supercritical fluid chromatography (SFC) and matrix-assisted laser desorption ionization time-of-flight mass spectrometry (MALDI-TOF-MS). The homologous series of polypropyleneglycol diacrylate (DA), polypropyleneglycol monoacrylate (MA), and unreacted polypropyleneglycol (PG) were observed as Na adducts in the MALDI-MS spectra of the PGA samples. The relative intensities of these peaks reflect the distributions of the homologues, although their accurate quantification was generally difficult because of change in the ionization efficiency depending on the chemical structure and the molecular weight of the species. On the other hand, the DA and the MA homologues were observed in the chromatograms obtained by SFC in a temperature-programming mode, while the PG homologues were not detected under the given SFC conditions using UV detection. Here, the determination of the degree of polymerization of each component in the chromatograms was accomplished through SFC fractionation for the corresponding peaks, followed again by MALDI-TOF-MS measurement. Furthermore, most of the components in the PGA samples were almost completely separated in the resulting gas chromatograms, and their unequivocal assignments were made also using the retention data on the gas chromatograms of the SFC fractions. As for the quantitative analysis, the relative abundances among DA, MA and PG for lower degrees of polymerization can be easily calculated based on the observed gas chromatograms, whereas the distribution of DA and MA can be estimated from the observed SFC data even for the relatively higher molecular weight fractions, which are generally difficult to determine accurately by GC because of their lower volatility. These results demonstrated that even the complex PGA samples were able to be characterized in detail by using GC, SFC and MALDI-TOF-MS complementarily. PMID- 11918186 TI - Simple and sensitive determination of citrinin in Monascus by GC-selected ion monitoring mass spectrometry. AB - A new method for the qualitative and quantitative analysis of citrinin in Monascus by gas-chromatography-selected ion monitoring (SIM) mass spectrometry has been developed. GC separation of citrinin in Monascus extract was achieved without the need for chemical derivatization, and could be detected as a single peak when the SIM mode selected 5 prominent fragmentations (m/z of 220, 205, 177, 105 and 91). The quantitative detection limit for citrinin was approximately 1 ppb. Finally, the GC-separated analyte from Monascus extract, at a retention time of 10.89 min, was examined by the method of pattern recognition by comparison with a citrinin standard. The results show that the 2 compounds had a 94% similarity when the SIM mode was used. PMID- 11918187 TI - Highly selective iodide membrane electrode based on a cerium salen. AB - A highly selective PVC membrane electrode based on a cerium-salen complex was prepared. The sensor displays an anti-Hofmeister selectivity sequence with a preference for iodide ion over many common organic and inorganic anions. The proposed electrode exhibits a near-Nernstian behavior over a wide concentration range (5.0 x 10(-2) - 8.0 x 10(-6) M) with a slope of 57.5 mV per decade, and a detection limit of 6.0 x 10(-6) M. The electrode has a very fast response time and can be used in the pH range of 3.0 - 1 1.0. It was applied, as an indicator electrode, in potentiometric titration of Ag+ ions. PMID- 11918188 TI - Application of dopamine as an electroactive ligand for the determination of aluminum in biological fluids. AB - Dopamine (3,4-dihydroxyphenylethylamine, DA) is applied as an electroactive chelant for indirect determination of aluminum (Al) in biological fluids. It is observed that the decrease of the differential pulse voltammetric (DPV) anodic peak current of DA is linear with the increase of Al concentration. Under optimum experimental conditions (pH 8.6, 2.0 x 10(-4) M DA, and 0.03 M NH4Ac-NH3 x H2O buffer solution), two linear ranges, 5.0 x 10(-8) - 4.0 x 10(-7) M and 4.0 x 10( 7) - 7.2 x 10(-6) M Al(III), are obtained. The detection limit of Al is 1.9 x 10( 8) M and the relative standard deviation for 4 x 10(-6) M Al(III) is 3.1% (N = 8). Many biologically active foreign species have been selected for interference. Excellent recoveries and accuracy have been obtained in the measurements of Al in biological samples such as synthetic renal dialysate, Ringer's solution, human whole blood, cerebrospinal fluid of demented patient, and urine of diabetic patient. The methodological principle that Al complexes with DA on the electroactive position result in the depression of electrochemical activities of DA has been verified by comparing both the electrochemical behaviors and the spectroscopic responses like UV-vis and Raman of DA in the presence and in the absence of Al. PMID- 11918189 TI - Perspectives of carbon paste electrodes in stripping potentiometry. AB - The testing of various types of carbon paste electrodes (CPEs) for their use in stripping potentiometry was carried out by means of specially proposed procedures and by using various model analyte systems. CPEs containing three different pasting liquids (paraffin oil, silicone oil, tricresyl phosphate) were tested as supports for mercury- and gold films, and as substrates for electrolytic, adsorptive, ion-pair forming, and extractive accumulation. Test measurements in organic solvent-containing media and studies of some irreversible electrochemical reactions were performed as well. Individual examples to be studied allowed one to formulate and outline some perspectives of CPEs in potentiometric stripping analysis and related constant current stripping analysis. PMID- 11918190 TI - Liquid-liquid extraction of palladium(II) from hydrobromic acid media by hexadecylpyridinium bromide. AB - A simple and rapid liquid-liquid extraction of palladium has been studied involving ion-pairing of bromocomplexes of palladium(II) with hexadecylpyridinium bromide (HDPB) dissolved in chloroform. The stoichiometry and distribution of (HDP)2PdBr4 between the aqueous and organic phase was investigated by spectrophotometric mole ratio method. The extraction efficiency of palladium(II) by HDPB was studied as a function of several variables: acid, salt, surfactant concentration and equilibrium time. The results showed that PdBr4(2-) extraction could be explained by assuming the formation of (HDP)2PdBr4 complexes in the aqueous solution and transfer to organic phase. The extraction was fast and the shaking time was only a few min. The average recovery of palladium(II) from an aqueous solution containing 10 microg/ml of analyte was 99% with an RSD% of 0.95. The percentage recovery of 0.2 microg/ml palladium(II) was 96%. PMID- 11918191 TI - Comparison of heating extraction procedures for Al, Ca, Mg, and Mn in tea samples. AB - A focused-microwave-assisted procedure was adopted for the extraction of Al, Ca, Mg, and Mn in tea leaves. The efficiency of extraction was evaluated using diluted acids and a water-soluble alkaline tertiary-amines solution. The extraction procedure was implemented in 5 min. A conventional hot-plate digestion procedure was employed for a comparison. A colorless digest was obtained for all tea samples and the total contents determined for each analyte were employed for estimating the efficiency of extraction. Tea infusions were also prepared. Subsequent determinations of Al, Ca, Mg, and Mn were carried out using flame atomic absorption spectrophotometry (FAAS) and inductively coupled plasma optical emission spectrometry (ICP-OES). In most cases, quantitative, or at least semi quantitative, extractions were attained for Ca, Mg, and Mn. Lower recoveries were attained to Al, which seems to be related to compounds that bind this element in the sample matrix. Large variations in the trace-element composition of teas were observed; these results are discussed with reference to both extraction media and type of tea. PMID- 11918192 TI - Synergic extraction of lanthanoids by mixtures of LIX 54 (high molecular weight beta-diketone) and bidentate neutral amines. AB - The synergic extraction of lanthanoids has been investigated using mixtures of high molecular weight beta-diketone, LIX 54 (HA; major component, 1-phenyl-3 isoheptyl-1,3-propanedion) and bidentate neutral ligands (S) in toluene. The distribution behavior of bidentate amines (2,2'-bipyridine (bpy), 1,10 phenanthroline (phen) and 2,9-dimethyl-1,10-phenanthroline (dmp)) was investigated and their related equilibrium constants were evaluated. The synergic effect is produced by the formation of such adduct complexes as MA3 x S. The extraction constants with LIX 54 alone and those in the presence of bidentate ligands were determined for lanthanoid-series elements. PMID- 11918194 TI - Catalytic determination of traces of silver(I) using the oxidation of Janus Green with peroxodisulfate. AB - A method for sensitive and selective determination of silver based on the catalytic effect of silver(I) ion on the oxidation of Janus Green by peroxodisulfate is described. o-Phenanthroline is used as an activator. The rate of the decrease in absorbance of Janus Green (at 615 nm) is proportional to the concentration of silver in the range of 0.3-4.0 ng mL(-1) and 4.0-500.0 ng mL( 1). The theoretical limit of detection was 0.25 ng mL(-1). The method is free from most interferences. The method was applied to the determination of silver in plants (the uptake of silver by plants), in photographic solutions, lake water and several synthetic samples. PMID- 11918193 TI - Determination of trace lead in chinese herbs by derivative flame atomic absorption spectrometry using an atom-trapping technique. AB - A simple and sensitive method is described for the determination of trace lead in Chinese herbs by derivative atom trapping flame atomic absorption spectrometry (D AT-FAAS) with a modified water-cooled quartz atom-trapping tube. The effects that influence the sensitivity of the derivative method, such as the trap position, the flame conditions and the collection time, were studied. The characteristics of the derivative atom trapping-atomic absorption signal and the linear nature of the working curve were investigated. The sensitivity of the derivative method is 2 or 3 orders of magnitude higher than that of FAAS, and the detection limit improved by 1 or 2 orders of magnitude. Satisfactory recoveries of 93.0-108.0% for lead were obtained by determining several Chinese herbs with a relative standard derivation range of 2.9 to 4.2%. PMID- 11918196 TI - Effects of polymerization and hydrolysis degrees of poly(vinyl alcohol) (PVA) on the efficiency of the PVA-solid substrate for room-temperature phosphorimetry. AB - Several kinds of poly(vinyl alcohols) (PVAs) having different degrees of polymerization and hydrolysis were tested as a material of a solid substrate for room-temperature phosphorimetry (RTP). Effects of these differences on the efficiency of the solid substrate were investigated. Completely hydrolyzed PVAs acquired a luminescence property in the grinding process of substrate preparation, but partially hydrolyzed PVAs did not acquire this property. When the completely hydrolyzed PVA substrates were prepared by drying their aqueous solutions, their luminescence property almost disappeared. However, very weak background emission remained on the surface of a completely dried substrate which had been treated with an analyte aqueous solution. This residual background affected the spectrum of the analyte, especially at low concentrations. Stability of the phosphorescence intensity with the passage of time was superior on the partially hydrolyzed PVAs than on the completely hydrolyzed PVAs. On the other hand, the RTP intensity and reproducibility were superior on the completely hydrolyzed PVAs. Practically, partially hydrolyzed PVAs were more suitable as a material of the substrate because of the stability of its RTP intensity and the weakness of its background emission. The linear dynamic range of the analytical curve for p-aminobenzoic acid on the substrate of partially hydrolyzed PVA having a degree of polymerization of 3,500 was 5-2,000 pmol/spot (20 microL) and its correlation coefficient was 0.963 for 30 data points. PMID- 11918195 TI - Spectrofluorometric resolution of closely overlapping drug mixtures using chemometrics methods. AB - A modified parallel factors analysis (PARAFAC) algorithm with a penalty diagonalization error (PDE), newly proposed by the present authors, was utilized to simultaneously resolve drug mixtures of propranolol (PRO), dipyridamole (DIP) and amiloride (AMI) without any loss of sensitivity. The analyses were performed in aqueous solution. The experimental results demonstrated that the profiles of the spectra and the concentrations could be accurately resolved using the PDE algorithm with a high sensitivity and stable repeatability. That is to say, the closely overlapping problem of the spectra could be easily solved. Furthermore, simultaneous determinations of three kinds of tablets, which contain PRO, AMI and DIP, respectively, were successfully performed with satisfactory results. PMID- 11918197 TI - Ion chromatographic determination of inorganic anions in environmental samples of Korea. PMID- 11918198 TI - Concentration on glass separatory funnel wall of anionic surfactants by ion association with methylene blue. PMID- 11918199 TI - On-line preconcentration and determination of traces of lead in river-water and seawater by flow injection-flame atomic absorption spectrometry and ICP-mass spectrometry. PMID- 11918200 TI - Spectrophotometric determination of nitrate in polluted water using a new coupling reagent. PMID- 11918201 TI - Preparation of chitosan derivatives containing methylthiocarbamoyl and phenylthiocarbamoyl groups and their selective adsorption of copper(II) over iron(III). PMID- 11918202 TI - Direct electrospray ionization mass spectroscopic measurement of micro-flow oil/water system. PMID- 11918203 TI - Crystal structure of methyl (8R,11R,14S)-8-[N-[(tert-butoxy)carbonyl]amino]11 (3,5-dichloro-4-hydroxyphenyl)-18,19-dibromo-5-iodo-4-hydroxy-13methyl-9,12-dioxo 10,13-diaza-2-oxatricyclo[14.2.2.1(3,7)]heneicosa-3,5,7(21), 16,18,19-hexaene-14 carboxylate. PMID- 11918204 TI - Crystal structure of o-(p-N,N-dimethylaminobenzoyl)benzoic acid. PMID- 11918205 TI - Crystal structure of 1,1,1-triphenyl-4-(10-benz[a]azulenyl)-3,4-buten-2-one. PMID- 11918207 TI - Crystal structure of N,N'-bis(4-hydroxysalicylidene)-1,2-phenylenediimine, methanol solvate. PMID- 11918206 TI - Crystal structure of [N,N'-bis(3,5-dinitrosalicylidene)-1,3-propanediaminato) bis(3,4-dimethylpyridine)]nickel(II)dioxane solvate. PMID- 11918208 TI - Mechanisms for lateral turns in lamprey in response to descending unilateral commands: a modeling study. AB - Straight locomotion in the lamprey is, at the segmental level, characterized by alternating bursts of motor activity with equal duration and spike frequency on the left and the right sides of the body. Lateral turns are characterized by three main changes in this pattern: (1) in the turn cycle, the spike frequency, burst duration, and burst proportion (burst duration/cycle duration) increase on the turning side; (2) the cycle duration increases in both the turn cycle and the succeeding cycle; and (3) in the cycle succeeding the turn cycle, the burst duration increases on the non-turning side (rebound). We investigated mechanisms for the generation of turns in single-segment models of the lamprey locomotor spinal network. Activation of crossing inhibitory neurons proved a sufficient mechanism to explain all three changes in the locomotor rhythm during a fictive turn. Increased activation of these cells inhibits the activity of the opposite side during the prolonged burst of the turn cycle, and slows down the locomotor rhythm. Secondly, this activation of the crossing inhibitory neurons is accompanied by an increased calcium influx into the cells. This gives a suppressed activity on the turning side and a contralateral rebound after the turn, through activation of calcium-dependent potassium channels. PMID- 11918209 TI - From view cells and place cells to cognitive map learning: processing stages of the hippocampal system. AB - The goal of this paper is to propose a model of the hippocampal system that reconciles the presence of neurons that look like "place cells" with the implication of the hippocampus (Hs) in other cognitive tasks (e.g., complex conditioning acquisition and memory tasks). In the proposed model, "place cells" or "view cells" are learned in the perirhinal and entorhinal cortex. The role of the Hs is not fundamentally dedicated to navigation or map building, the Hs is used to learn, store, and predict transitions between multimodal states. This transition prediction mechanism could be important for novelty detection but, above all, it is crucial to merge planning and sensory-motor functions in a single and coherent system. A neural architecture embedding this model has been successfully tested on an autonomous robot, during navigation and planning in an open environment. PMID- 11918210 TI - Understanding finger coordination through analysis of the structure of force variability. AB - Most common motor acts involve highly redundant effector systems. Understanding how such systems are controlled by the nervous system is a long-standing scientific challenge. Most proposals for solving this problem are based on the assumption that a particular solution, which optimizes additional constraints, is selected by the nervous system out of the many possible solutions. This study attempts to address this question in the context of coordinating individual finger forces to produce a controlled total force oscillation between 5% and 35% of each subject's maximum force of voluntary contraction, under two different combinations of four fingers. The structure of variability of individual finger forces was evaluated with respect to hypotheses that, at each instance in time, subjects attempt to: (1) stabilize the value of total force and (2) stabilize the total moment created by the fingers about the long axis passing through the forearm and midline of the hand. The results provide evidence that a range of goal-equivalent finger force combinations is generated to stabilize the values of total force and the total moment. The control of total force was specified explicitly by the task. However, it was stabilized only near the time of peak force. In contrast, the total moment was stabilized throughout most of the force cycle. The results lead to the suggestion that successful task performance is achieved, not by selecting a single optimal solution, but by discovering an appropriate control law that selectively stabilizes certain combinations of degrees of freedom relevant to the task while releasing from control other combinations. PMID- 11918211 TI - Modeling visuospatial perception in neglect patients. AB - The spatial distortion hypothesis is one of several theories that explain certain aspects of neglect in patients with right parietal lesions. To determine whether a distorted representation of space can account for the performance of neglect patients in different visuospatial tasks, we asked 26 neglect patients to: (1) bisect horizontal lines and (2) to compare the width of two horizontally aligned bars. A simple mathematical model compatible with the idea of a stationary distortion of represented space in egocentric coordinates explained the results of the line-bisection task. A second model that had basically the same structure and was compatible with the idea of a distorted egocentric representation based on a dynamic remapping of space approximated the size-comparison data. These results support the view that abnormalities observed in the line-bisection and size-comparison tasks are due to a distorted internal representation of the external world. Certain findings suggest that this distortion could be based on a dynamic mapping of space determined by the distribution of visuospatial attention. PMID- 11918212 TI - Linear combinations of nonlinear models for predicting human-machine interface forces. AB - This study presents a computational framework that capitalizes on known human neuromechanical characteristics during limb movements in order to predict human machine interactions. A parallel-distributed approach, the mixture of nonlinear models, fits the relationship between the measured kinematics and kinetics at the handle of a robot. Each element of the mixture represented the arm and its controller as a feedforward nonlinear model of inverse dynamics plus a linear approximation of musculotendonous impedance. We evaluated this approach with data from experiments where subjects held the handle of a planar manipulandum robot and attempted to make point-to-point reaching movements. We compared the performance to the more conventional approach of a constrained, nonlinear optimization of the parameters. The mixture of nonlinear models accounted for 79 +/- 11% (mean +/- SD) of the variance in measured force, and force errors were 0.73 +/- 0.20% of the maximum exerted force. Solutions were acquired in half the time with a significantly better fit. However, both approaches suffered equally from the simplifying assumptions, namely that the human neuromechanical system consisted of a feedforward controller coupled with linear impedances and a moving state equilibrium. Hence, predictability was best limited to the first half of the movement. The mixture of nonlinear models may be useful in human-machine tasks such as in telerobotics, fly-by-wire vehicles, robotic training, and rehabilitation. PMID- 11918213 TI - Two different pathways for the maintenance of trabecular bone in adult male mice. AB - Androgens may regulate the male skeleton either directly via activation of the androgen receptor (AR) or indirectly via aromatization of androgens into estrogen and, thereafter, via activation of estrogen receptors (ERs). There are two known estrogen receptors, ER-alpha and ER-beta. The aim of this study was to investigate the relative roles of ER-alpha, ER-beta, and AR in the maintenance of trabecular bone in male mice. Seven-month-old male mice, lacking ER-alpha (ERKO), ER-beta (BERKO), or both receptors (DERKO), were orchidectomized (orx) and treated for 3 weeks with 0.7 microg/mouse per day of 17beta-estradiol or vehicle. No reduction in trabecular bone mineral density (BMD) was seen in ERKO, BERKO, or DERKO mice before orx, showing that neither ER-a nor ER-beta is required for the maintenance of a normal trabecular BMD in male mice. After orx, there was a pronounced decrease in trabecular BMD, similar for all groups, resulting in equal levels of trabecular BMD in all genotypes. This reduction was reversed completely in wild-type (WT) and BERKO mice treated with estrogen, and no significant effect of estrogen was found in ERKO or DERKO mice. In summary, the trabecular bone is preserved both by a testicular factor, presumably testosterone acting via AR and by an estrogen-induced activation of ER-alpha. These results indicate that AR and ER-alpha are redundant in the maintenance of the trabecular bone in male mice. In contrast, ER-beta is of no importance for the regulation of trabecular bone in male mice. PMID- 11918214 TI - Sex steroids, mice, and men: when androgens and estrogens get very close to each other. PMID- 11918215 TI - Generation of a new congenic mouse strain to test the relationships among serum insulin-like growth factor I, bone mineral density, and skeletal morphology in vivo. AB - Insulin-like growth factor (IGF) I is a critical peptide for skeletal growth and consolidation. However, its regulation is complex and, in part, heritable. We previously indicated that changes in both serum and skeletal IGF-I were related to strain-specific differences in total femoral bone mineral density (BMD) in mice. In addition, we defined four quantitative trait loci (QTLs) that contribute to the heritable determinants of the serum IGF-I phenotype in F2 mice derived from progenitor crosses between C3H/HeJ (C3H; high total femoral BMD and high IGF I) and C57BL/6J (B6; low total femoral BMD and low IGF-I) strains. The strongest QTL, IGF-I serum level 1 (Igflsl-1; log10 of the odds ratio [LOD] score, approximately 9.0), is located on the middle portion of chromosome (Chr) 6. For this locus, C3H alleles are associated with a significant reduction in serum IGF I. To test the effect of this QTL in vivo, we generated a new congenic strain (B6.C3H-6T [6T]) by placing the Chr 6 QTL region (D6Mit93 to D6Mit150) from C3H onto the B6 background. We then compared serum and skeletal IGF-I levels, body weight, and several skeletal phenotypes from the N9 generation of 6T congenic mice against B6 control mice. Female 6T congenic mice had 11-21% lower serum IGF I levels at 6, 8, and 16 weeks of age compared with B6 (p < 0.05 for all). In males, serum IGF-I levels were similar in 6T congenics and B6 controls at 6 weeks and 8 weeks but were lower in 6T congenic mice at 16 weeks (p < 0.02). In vitro, there was a 40% reduction in secreted IGF-I in the conditioned media (CMs) from 6T calvaria osteoblasts compared with B6 cells (p < 0.01). Total femoral BMD as measured by peripheral quantitative computed tomography (pQCT) was lower in both 6T male (-4.8%, p < 0.01) and 6T female (-2.3%, p = 0.06) congenic mice. Geometric features of middiaphyseal cortical bone were reduced in 6T congenic mice compared with control mice. Femoral cancellous bone volume (BV) density and trabecular number (Tb.N) were 50% lower, whereas trabecular separation (Tb.Sp) was 90% higher in 8-week-old female 6T congenic mice compared with B6 control mice (p < 0.01 for all). Similarly, vertebral cancellous BV density and Tb.N were lower (-29% and -19%, respectively), whereas Tb.Sp was higher (+29%) in 16-week old female 6T congenic mice compared with B6 control mice (p < 0.001 for all). Histomorphometric evaluation of the proximal tibia indicated that 6T congenics had reduced BV fraction, labeled surface, and bone formation rates compared with B6 congenic mice. In summary, we have developed a new congenic mouse strain that confirms the Chr 6 QTL as a major genetic regulatory determinant for serum IGF-I. This locus also influences bone density and morphology, with more dramatic effects in cancellous bone than in cortical bone. PMID- 11918216 TI - Estrogen receptor isoform-specific induction of progesterone receptors in human osteoblasts. AB - Estrogen induction of progesterone receptor (PR) expression may be important to bone physiology because progesterone has been implicated in the control of bone formation and resorption. Although PR gene expression can be induced in osteoblasts by estrogen signaling through the estrogen receptor (ER) a isoform, it is unknown whether the ER-beta isoform is involved in this regulation. The effect of estrogen on PR expression was examined in human fetal osteoblast (hFOB) cell lines stably transfected with either ER-alpha or ER-beta. Estrogen treatment of hFOB/ER-a cells induced PR messenger RNA (mRNA) steady-state levels after 24 h and protein levels after 48 h, as established by competitive reverse transcriptase-polymerase chain reaction (RT-PCR) and Western blotting. Interestingly, no induction of PR expression was observed in the hFOB/ER-beta cells during this period. However, PR mRNA was induced progressively after 48 h of treatment with estrogen with maximum levels achieved at 12 days posttreatment. ER protein also was increased after 12 days of treatment. Both A and B isoforms of PR (PRA and PRB) were induced by estrogen in the hFOB/ER-a cells as well as much later in hFOB/ER-beta cells. The pure antiestrogen ICI 182,780 prevented PR induction by estrogen in both cell lines. An ER-beta-selective antagonist R, R tetrahydrochrysene (THC) abolished the induction of PR mRNA in hFOB/ER-beta but not in hFOB/ER-a cells, verifying that the response in the former cell line was ER-beta-mediated. Transient cotransfection of hFOB cells with ER-a or ER-beta together with either a human PRA or PRB promoter linked to a reporter plasmid revealed that although the PRB promoter was stimulated equally by estrogen activation of either ER isoform, PRA was activated preferentially by ER-alpha. Together, these results show that although estrogen can up-regulate endogenous PR gene expression in osteoblasts via both ER isoforms, ER-alpha is the predominant inducer. PMID- 11918217 TI - The mode of mechanical integrin stressing controls intracellular signaling in osteoblasts. AB - Following the idea that integrin receptors function as mechanotransducers, we applied defined physical forces to integrins in osteoblastic cells using a magnetic drag force device to show how cells sense different modes of physical forces. Application of mechanical stress to the beta1-integrin subunit revealed that cyclic forces of 1 Hz were more effective to stimulate the cellular calcium response than continuous load. Cyclic forces also induced an enhanced cytoskeletal anchorage of tyrosine-phosphorylated proteins and increased activation of the focal adhesion kinase (FAK) and mitogen activated protein (MAP) kinase. These events were dependent on an intact cytoskeleton and the presence of intracellular calcium. Analyses of the intracellular spatial organization of the calcium responses revealed that calcium signals originate in a restricted region in the vicinity of the stressed receptors, which indicates that cells are able to sense locally applied stress on the cell surface via integrins. The calcium signals can spread throughout the cell including the nucleus, which shows that calcium also is a candidate to transmit mechanically induced information into different cellular compartments. PMID- 11918218 TI - Possible involvement of IkappaB kinase 2 and MKK7 in osteoclastogenesis induced by receptor activator of nuclear factor kappaB ligand. AB - Recent studies have revealed the essential role of the receptor activator of nuclear factor kappaB (NF-kappaB) ligand (RANKL) in osteoclast differentiation and activation. Adenovirus vector could efficiently transduce genes into RAW264.7 cells, which differentiate into osteoclast-like multinucleated cells in the presence of RANKL. The role of NF-kappaB and c-jun N-terminal kinase (JNK) activation in RANKL-induced osteoclast differentiation was investigated using an adenovirus vector carrying the dominant negative 1kappaB kinase 2 gene (AxIKK2DN) or dominant negative MKK7 gene (AxMKK7DN). IKK2DN and MKK7DN overexpression in RAW cells specifically suppressed the NF-kappaB activation and JNK activation in response to RANKL, respectively, without affecting other signaling pathways. Either inhibition of NF-kappaB or JNK pathways dose-dependently inhibited osteoclast formation induced by RANKL. These results suggest that both NF-kappaB and JNK activation are independently required for osteoclast differentiation. PMID- 11918219 TI - Vitamin D hormone inhibits osteoclastogenesis in vivo by decreasing the pool of osteoclast precursors in bone marrow. AB - Previous observations that vitamin D hormone induces the expression of the receptor activator of nuclear factor kappaB (NF-kappaB) ligand (RANKL), thereby stimulating osteoclastogenesis in vitro, led to the widespread belief that 1alpha,25-dihydroxyvitamin D3 [1a,25(OH)2D3] is a bone-resorbing hormone. Here, we show that alfacalcidol, a prodrug metabolized to 1alpha,25(OH)2D3, suppresses bone resorption at pharmacologic doses that maintain normocalcemia in an ovariectomized (OVX) mouse model of osteoporosis. Treatment of OVX mice with pharmacologic doses of alfacalcidol does not increase RANKL expression, whereas toxic doses that cause hypercalcemia markedly reduce the expression of RANKL. When bone marrow (BM) cells from OVX mice were cultured with sufficient amounts of macrophage colony-stimulating factor (M-CSF) and RANKL, osteoclastogenic activity was higher than in sham mice. Marrow cultures from alfacalcidol- or estrogen-treated OVX mice showed significantly less osteoclastogenic potential compared with those from vehicle-treated OVX mice, suggesting that the pool of osteoclast progenitors in the marrow of vitamin D-treated mice as well as estrogen-treated mice was decreased. Frequency analysis showed that the number of osteoclast progenitors in bone marrow was increased by OVX and decreased by in vivo treatment with alfacalcidol or estrogen. We conclude that the pharmacologic action of active vitamin D in vivo is to decrease the pool of osteoclast progenitors in BM, thereby inhibiting bone resorption. Because of its unusual activity of maintaining bone formation while suppressing bone resorption, in contrast to estrogens that depress both processes, vitamin D hormone and its bone selective analogs may be useful for the management of osteoporosis. PMID- 11918220 TI - Down-regulation of osteoclast differentiation by daidzein via caspase 3. AB - Phytoestrogens are plant-derived compounds with estrogen-like activity. Phytoestrogen-rich diets may prevent postmenopausal osteoporosis and these molecules maintain bone mass in ovariectomized animals. We compared the effects of the isoflavone daidzein, which has no action on tyrosine kinases, and 17beta estradiol on the development and activity of osteoclasts in vitro. Nonadherent porcine bone marrow cells were cultured on dentine slices or on culture slides in the presence of 10-8 M of 1,25-dihydroxyvitamin D3 [1,25(OH)2D3], with or without 10(-8) M of daidzein, 10(-8) M of 17beta-estradiol for 9-11 days. Multinucleated tartrate-resistant acid phosphatase-positive (TRAP+) cells that resorbed bone (osteoclasts) developed in the presence of 1,25(OH)2D3. The number of osteoclasts formed in response to 1,25(OH)2D3 was reduced by 58 +/- 8% by daidzein and 52 +/- 5% by estrogen (p < 0.01); these effects were reversed by 10-6 M of ICI 182,780. The area resorbed by mature osteoclasts was reduced by 39 +/- 5% by daidzein and 42 +/- 6% by estradiol (p < 0.01). Both compounds also inhibited the 1,25(OH)2D3 induced differentiation of osteoclast progenitors (mononucleated TRAP+ cells), 53 +/- 8% by daidzein and 50 +/- 7% by estradiol (p < 0.05). Moreover, daidzein and estradiol promoted caspase-8 and caspase-3 cleavage and DNA fragmentation of monocytic bone marrow cells. Caspase-3 cleavage was reversed by 10-8 M of ICI 182,780. Both compounds up-regulated the expression of nuclear estrogen receptors ER-alpha and ER-beta. Thus, daidzein, at the same concentration as 17beta estradiol, inhibits osteoclast differentiation and activity. This may be caused by, at least in part, greater apoptosis of osteoclast progenitors mediated by ERs. PMID- 11918221 TI - Proteolysis involving matrix metalloproteinase 13 (collagenase-3) is required for chondrocyte differentiation that is associated with matrix mineralization. AB - Collagenases are involved in cartilage matrix resorption. Using bovine fetal chondrocytes isolated from physeal cartilages and separated into a distinct prehypertrophic subpopulation, we show that in serum-free culture they elaborate an extracellular matrix and differentiate into hypertrophic chondrocytes. This is characterized by expression of type X collagen and the transcription factor Cbfal and increased incorporation of 45Ca2+ in the extracellular matrix, which is associated with matrix calcification. Collagenase activity, attributable only to matrix metalloproteinase (MMP) 13 (collagenase-3), is up-regulated on differentiation. A nontoxic carboxylate inhibitor of MMP-13 prevents this differentiation; it suppresses expression of type X collagen, Cbfal, and MMP-13 and inhibits increased calcium incorporation in addition to inhibiting degradation of type II collagen in the extracellular matrix. General synthesis of matrix proteins is unaffected. These results suggest that proteolysis involving MMP-13 is required for chondrocyte differentiation that occurs as part of growth plate development and which is associated with matrix mineralization. PMID- 11918222 TI - Cartilage calcification studied by proton nuclear magnetic resonance microscopy. AB - A three-dimensional (3D) mineralizing culture system using hollow fiber bioreactors has been developed to study the early stages of endochondral ossification by proton nuclear magnetic resonance (NMR) microscopy. Chondrocytes harvested from the cephalic half of the sterna from 17-day-old chick embryos were terminally differentiated with 33 nM of retinoic acid for 1 week and mineralization was initiated by the addition of 1% beta-glycerophosphate to the culture medium. Histological sections taken after 6 weeks of development in culture confirmed calcification of the cartilage matrix formed in bioreactors. Calcium to phosphorus ratios (1.62-1.68) from X-ray microanalysis supported electron diffraction of thin tissue sections showing the presence of a poorly crystalline hydroxyapatite mineral phase in the cultures. After 4 weeks of culture, quantitative proton NMR images showed water proton magnetization transfer rate constants (km) were higher in premineralized cartilage compared with uncalcified cartilage, a result suggesting collagen enrichment of the matrix. Notably after 5 weeks mineral deposits formed in bioreactors principally in the collagen-enriched zones of the cartilage with increased km values. This caused marked reductions in water proton longitudinal (T1) and transverse (T2) relaxation times and water diffusion coefficients (D). These results support the hypothesis that mineralization proceeds in association with a collagen template. After 6 weeks of culture development, the water proton T2 values decreased by 13% and D increased by 7% in uncalcified areas, compared with the same regions of tissue examined 1 week earlier. These changes could be attributed to the formation of small mineral inclusions in the cartilage, possibly mediated by matrix vesicles, which may play an important role in cartilage calcification. In summary, NMR images acquired before and after the onset of mineralization of the same tissue provide unique insights into the matrix events leading to endochondral mineral formation. PMID- 11918223 TI - Resistance to unloading-induced three-dimensional bone loss in osteopontin deficient mice. AB - Recent development in three-dimensional (3D) imaging of cancellous bone has made possible true 3D quantification of trabecular architecture. This provides a significant improvement in the measures available to study and understand the mechanical functions of cancellous bone. We recently reported that the presence of osteopontin (OPN) was required for the effects of mechanical stress on bone as OPN-null (OPN-/-) mice showed neither enhancement of bone resorption nor suppression of bone formation when they were subjected to unloading by tail suspension. However, in this previous study, morphological analyses were limited to two-dimensional (2D) evaluation. Although bone structure is 3D and thus stress effect should be evaluated based on 3D parameters, no such 3D morphological features underlying the phenomenon have been known. To elucidate the role of OPN in mediating mechanical stress effect based on true quantitative examination of bone, we evaluated 3D trabecular structures of hindlimb bones of OPN-/- mice after tail suspension. Tail suspension significantly reduced 3D parameters of bone volume (BV/TV), trabecular number (Tb.N), trabecular thickness (Tb.Th), and anisotropy and increased 3D parameters on trabecular separation (Tb.Sp) in wild type mice. In contrast, these 3D parameters were not altered after tail suspension in OPN-/- mice. These data provided evidence that OPN is required for unloading-induced 3D bone loss. PMID- 11918224 TI - Transforming growth factor beta2 inhibits adipocyte differentiation induced by skeletal unloading in rat bone marrow stroma. AB - Skeletal unloading induced by hindlimb suspension in rats reduces bone formation and induces osteopenia, but its effect on adipogenesis is unknown. We assessed the effects of unloading and transforming growth factor (TGF) beta2 on bone marrow stromal cell adipocyte differentiation in relation with osteoblast differentiation. Skeletal unloading rapidly (4-7 days) decreased osteoblast transcription factor Runx2, osteocalcin (OC), and type I collagen messenger RNA (mRNA) levels and reduced bone formation in the long bone metaphysis. Conversely, unloading increased expression of the adipocyte transcription factor peroxisome proliferator-activated receptor gamma2 (PPARgamma2) at 4 days and increased expression of the adipocyte differentiation genes lipoprotein lipase (LPL) and aP2 in the bone marrow stroma at 7 days. Consistently, unloading increased the number and volume of adipocytes in the bone marrow stroma. Continuous (0-7 days) and late (4-7 days) treatments with TGF-beta2 corrected the abnormal expression of Cbfa1/Runx2, OC, and type I collagen mRNAs and normalized bone formation in unloaded metaphyseal bone. Moreover, both TGF-beta2 treatments decreased PPARy2 and C/EBPalpha mRNA levels at 4 days and normalized aP2 and LPL expression and adipocyte number and volume at 7 days. These results show that skeletal unloading increases adipocyte differentiation concomitantly with inhibition of osteoblast differentiation. These abnormalities are prevented and reversed by TGF-beta2, suggesting a role for TGF-beta in the control of adipogenic differentiation in the bone marrow stroma. PMID- 11918226 TI - Mechanical and architectural bone adaptation in early stage experimental osteoarthritis. AB - The purpose of this study was to quantify mechanical and architectural changes to knee joint periarticular subchondral cancellous bone in early stage experimental osteoarthritis (OA). Unilateral anterior cruciate ligament transection (ACLX) was performed on 10 dogs that were assigned randomly to two groups: 3 weeks or 12 weeks post-ACLX. Cylindrical bone cores excised from the medial condyle of the distal femur after death were scanned using high-resolution microcomputed tomography (muCT) and subsequently failed under unconstrained uniaxial compression. The apparent-level elastic modulus was less in the ACLX femur compared with the contralateral control, and the decrease was significant (-45%; p < 0.05) by 12-weeks post-ACLX. A finite element (FE) analysis based on muCT data simulated the uniaxial compression tests on a specimen-by-specimen basis to determine tissue modulus. No change in tissue modulus was detected, and a single tissue modulus of 5100 MPa (95% CI, +/- 600 MPa) explained the apparent-level modulus changes observed in the disease-related bone adaptation. The three dimensional (3D) connectivity was evaluated from the original muCT data to quantify architectural alterations in contrast to tissue alterations. Significantly increased connectivity (through plate perforations) occurred as early as 3 weeks post-ACLX and was as high as 127% by 12 weeks post-ACLX in the distal femur. These measured changes indicated that architectural adaptation predominated over tissue modulus changes affecting apparent-level elastic modulus in the early stage of experimental OA and suggests that to maintain normal cancellous bone after a traumatic injury, early intervention should focus on preventing the substantial architectural alterations. PMID- 11918225 TI - Tests of linkage and/or association of genes for vitamin D receptor, osteocalcin, and parathyroid hormone with bone mineral density. AB - Bone mineral density (BMD) is a major determinant of osteoporotic fractures (OFs). The heritability of BMD ranges from 50% to 90% in human populations. Extensive molecular genetic analyses have been performed through traditional linkage or association approaches to test and identify genes or genomic regions underlying BMD variation. The results, particularly those concerning the vitamin D receptor (VDR) gene, have been inconsistent and controversial. In this study, we simultaneously test linkage and/or association of the genes for VDR, osteocalcin (also known as bone Gla protein [BGP]), and parathyroid hormone (PTH) with BMD in 630 subjects from 53 human pedigrees. Each of these pedigrees was ascertained through a proband with an extreme BMD value at the hip or spine (Z score < or = -1.28). For the raw BMD values, adjusting for significant covariate effects of age, sex, and weight, we performed tests for linkage alone, association alone, and then both linkage and association. For the spine BMD, at the two markers (ApaI and FokI) inside the VDR gene we found evidence for linkage (p < 0.05) and for both linkage and association by the transmission disequilibrium test (TDT; p < 0.05); association was detected (p < 0.07) with regular statistical testing by analyses of variance (ANOVA). In addition, significant results were found for association alone (p < 0.05), linkage alone (p = 0.0005), and for linkage and association (p = 0.0019) for the intragenic marker HindIII of the BGP gene for the hip BMD. Through testing for association, linkage, and linkage and association simultaneously, our data support the VDR gene as a quantitative trait locus (QTL) underlying spine BMD variation and the BGP gene as a QTL underlying hip BMD variation. However, our data do not support the PTH gene as a QTL underlying hip or spine BMD variation. This is the first study in the broad field of bone genetics that tests candidate genes as QTLs for BMD by testing simultaneously for association alone, for linkage alone, and for association and linkage (via the TDT). PMID- 11918227 TI - Effect of Crohn's disease on bone metabolism in vitro: a role for interleukin-6. AB - Circulating proinflammatory cytokines may be involved in osteopenia associated with Crohn's disease (CD). Therefore, the effect of interleukin (IL)-6, IL-1beta, and tumor necrosis factor (TNF) a contained in Crohn's serum on bone formation was examined in a bone organ culture system. Initially, serum levels of IL-6, IL 1beta, and TNF-a were determined by ELISA in newly diagnosed, untreated children with CD and healthy age-matched controls. Serum IL-6 levels were significantly higher in patients with CD than in controls (23.9 +/- 2.8 pg/ml vs. 0.7 pg/ml +/- 0.2; p < 0.001), whereas IL-1beta and TNF-alpha serum levels were not. In the organ culture studies, 20-day-old fetal rat parietal bones were incubated for 96 h with CD or control serum, serum preincubated with a neutralizing antibody to each cytokine or a nonimmune immunoglobulin control, and with IL-6. Bone formation measured by assaying calcium content and dry weight was significantly decreased in bones exposed to Crohn's serum. Light microscopy of the bones treated with CD serum revealed a discontinuous, uneven mineralized bone matrix and disorganized osteoblasts with altered morphology. Incubation with an antibody that neutralized IL-6 activity prevented the change in osteoblast and bone morphology. TNF-a and IL-1beta antibodies had no apparent effects. Collagen synthesis and DNA content were not affected by CD serum. Also, addition of IL-6 to the culture medium decreased mineralization. These results suggest that IL-6 is a mediator of the effects of Crohn's serum on in vitro mineralization and may be a contributing factor to the osteopenia associated with CD. PMID- 11918228 TI - Can vitamin D supplementation reduce the risk of fracture in the elderly? A randomized controlled trial. AB - Randomized controlled trials have shown that a combination of vitamin D and calcium can prevent fragility fractures in the elderly. Whether this effect is attributed to the combination of vitamin D and calcium or to one of these nutrients alone is not known. We studied if an intervention with 10 microg of vitamin D3 per day could prevent hip fracture and other osteoporotic fractures in a double-blinded randomized controlled trial. Residents from 51 nursing homes were allocated randomly to receive 5 ml of ordinary cod liver oil (n = 569) or 5 ml of cod liver oil where vitamin D was removed (n = 575). During the study period of 2 years, fractures and deaths were registered, and the principal analysis was performed on the intention-to-treat basis. Biochemical markers were measured at baseline and after 1 year in a subsample. Forty-seven persons in the control group and 50 persons in the vitamin D group suffered a hip fracture. The corresponding figures for all nonvertebral fractures were 76 persons (control group) and 69 persons (vitamin D group). There was no difference in the incidence of hip fracture (p = 0.66, log-rank test), or in the incidence of all nonvertebral fractures (p = 0.60, log-rank test) in the vitamin D group compared with the control group. Compared with the control group, persons in the vitamin D group increased their serum 25-hydroxyvitamin D concentration with 22 nmol/liter (p = 0.001). In conclusion, we found that an intervention with 10 microg of vitamin D3 alone produced no fracture-preventing effect in a nursing home population of frail elderly people. PMID- 11918229 TI - Incidence of vertebral fracture in europe: results from the European Prospective Osteoporosis Study (EPOS). AB - Vertebral fracture is one of the major adverse clinical consequences of osteoporosis; however, there are few data concerning the incidence of vertebral fracture in population samples of men and women. The aim of this study was to determine the incidence of vertebral fracture in European men and women. A total of 14,011 men and women aged 50 years and over were recruited from population based registers in 29 European centers and had an interviewer-administered questionnaire and lateral spinal radiographs performed. The response rate for participation in the study was approximately 50%. Repeat spinal radiographs were performed a mean of 3.8 years following the baseline film. All films were evaluated morphometrically. The definition of a morphometric fracture was a vertebra in which there was evidence of a 20% (+4 mm) or more reduction in anterior, middle, or posterior vertebral height between films--plus the additional requirement that a vertebra satisfy criteria for a prevalent deformity (using the McCloskey-Kanis method) in the follow-up film. There were 3174 men, mean age 63.1 years, and 3,614 women, mean age 62.2 years, with paired duplicate spinal radiographs (48% of those originally recruited to the baseline survey). The age standardized incidence of morphometric fracture was 10.7/1,000 person years (pyr) in women and 5.7/1,000 pyr in men. The age-standardized incidence of vertebral fracture as assessed qualitatively by the radiologist was broadly similar-12.1/1,000 pyr and 6.8/1,000 pyr, respectively. The incidence increased markedly with age in both men and women. There was some evidence of geographic variation in fracture occurrence; rates were higher in Sweden than elsewhere in Europe. This is the first large population-based study to ascertain the incidence of vertebral fracture in men and women over 50 years of age across Europe. The data confirm the frequent occurrence of the disorder in men as well as in women and the rise in incidence with age. PMID- 11918230 TI - Gender differences in the genetic factors responsible for variation in bone density and ultrasound. AB - Although genetic factors are thought to explain a large proportion of the variation in bone density in women, few studies have been conducted in men. Therefore, it is unclear whether the individual differences in bone strength between men and women are a reflection of gender differences in the relative influence of genetic and environmental factors on bone density variance. The aim of this study was to determine if there were gender differences in the genetic components of variance for bone density and ultrasound. In addition, the study aimed to explore the hypothesis that there are unique gender-specific genetic determinants of these traits. Bone mineral density (BMD) of the hip, distal forearm, and lumbar spine were measured by dual-energy X-ray absorptiometry (DXA) as well as quantitative ultrasound (QUS) at the calcaneus in healthy female twin pairs (286 identical [MZ] and 265 nonidentical [DZ]), male twin pairs (72 MZ and 65 DZ), and 82 opposite-sex (OS) pairs aged between 18 and 80 years. For hip BMD, distal forearm, and QUS measurements, the differences between MZ correlations and like-sex DZ correlations were similar for both sexes, suggesting little difference in the component of total variance explained by genetic factors between male and female twin pairs. However, correlations between OS twin pairs were lower than that of like-sex twin pairs, suggesting the possibility of unique gender-specific genetic effects. At the forearm, model fitting suggested a small gender difference in the magnitude of genetic variance as well as the presence of a unique gender-specific genetic variance component. Hip, lumbar spine, and QUS measurements were better explained by models that assumed no gender differences in genetic variance between the sexes, but the study had insufficient power to detect small differences in the genetic components of variance. The results of this study suggest that the proportion of bone strength variance explained by genetic factors is similar for men and women. However, at some regions there is evidence to suggest a gender-specific genetic component to the overall genetic variance. PMID- 11918231 TI - Universal standardization of forearm bone densitometry. AB - As part of an effort to quantify device-dependent differences in forearm bone density, 101 women, aged 20-80 years (approximately 16 women in each age decade), were scanned on six forearm bone densitometers: the Aloka DCS-600EX, the Hologic QDR-4500A, the Lunar PIXI, the Norland pDEXA, the Osteometer DTX-200, and the Pronosco X-posure System. Regression statistics are reported for all similar regions of interest (ROIs). However, comparisons were confounded because of large differences in the ROI size and placement. The number of ROIs reported for a single scan by each device varied from 1 to 12. The correlation coefficients ranged from 0.7 < r < 0.97, with the highest correlation coefficients and lowest SEs for comparisons between the most similar ROIs. Standardized units of bone mineral density are derived for distal (sdBMD), mid-(smBMD), and proximal (spBMD) ROTs that allow for comparable mean bone densities to be derived for patient populations. Five phantoms were scanned and characterized on five of the devices and the precision and mean values were reported. These phantom values will aid in the in vitro cross-calibration between manufacturers to recreate the presented in vivo relationships. Care should be exercised when using these equations for cross calibrating patient databases or pooling clinical data from different devices because the least significant differences detectable from measurements taken on two different machines can be increased substantially. PMID- 11918232 TI - Is weight loss in obese premenopausal women associated with a decline in bone mass? PMID- 11918233 TI - Evaluation of the prognostic factors after thymoma resection. AB - We discuss the prognostic factors of thymoma clinicopathologically. Regarding the survival rate by the clinical stage classification of Masaoka, significant correlation was made between stage I and stage III (P < 0.05) and stage I and stage IVa (P < 0.03). The tumor resectability was classified into complete and incomplete resection, and a significant difference was shown by the survival rate of the complete resection at P < 0.0001. Regarding the survival rate by the invasive organ of the tumor, significant correlation was made between no invasion and the great vessel invasion (P < 0.0004) and between invasion except for the great vessel and great vessel invasion (P < 0.004). As for the histological type, the tendency in which the epithelial cell type predominancy increased with the progress of the clinical stage was shown. A significant correlation was not shown in the evaluation by adjuvant therapy. However, recently we have done chemotherapy and/or radiotherapy periodically for invasive thymoma. PMID- 11918234 TI - Total pelvic exenteration: results from a multispecialty team approach to complex cancer surgery. AB - Total pelvic exenteration is often the only curative option for recurrent or locally advanced pelvic cancers, but it carries a high risk of mortality and morbidity. A dedicated multispecialty team operative approach may provide the expertise to perform this uncommon procedure with favorable outcomes. Data were analyzed from a prospectively collected computerized database. There were 14 patients (2 men; mean age, 54.6 +/- 3.6 years) with mainly cervical cancers, of which 71.4% were recurrent. Anesthetic time was 5 +/- 0.9 hours, intraoperative blood loss was 2.1 +/- 0.5 liters, and postoperative hospitalization was 22 +/- 9.9 days. An ileal conduit was performed in all patients, but intestinal continuity was restorable with colonic J-pouch in 71.4% of the patients. There was no mortality at 30 days or during hospitalization. Complication rates were 35.7%, accounting for reoperations in 28.6%. Recurrences were detected in 50% patients at a mean follow-up of 53.1 +/- 9.2 months. The mean time for cancer recurrence was 13.3 +/- 3.3 months. Fifty percent of those patients had otherwise survived to date. We conclude that a dedicated multispecialty team may perform total pelvic exenteration with minimum mortality and acceptable morbidity. PMID- 11918235 TI - Thyroid surgery according to Enderlen-Hotz or Dunhill: a comparison of two surgical methods for the treatment of Graves' disease. AB - The surgical therapy of Graves' disease is favored in Germany, which is an iodine deficient area, and nodular transformation is common. The surgical methods mainly differ in their extent of resection. We examined 152 patients with regard to operative and postoperative complications in a prospective randomized study. Patients were randomized to two groups: treated either by subtotal resection on both sides (STT) or subtotal resection with contralateral hemithyroidectomy (SHT), the two most common surgical procedures. The follow-up period was 6 years; complication and recurrence rates were studied. There was no significant difference concerning mortality, recurrent laryngeal nerve palsy, hypocalcemia, secondary hemorrhage, wound infections, or recurrence rate between STT and SHT. However, SHT only requires a unilateral resection in case of a recurrence, thus reducing the risk of bilateral recurrent laryngeal nerve palsy. PMID- 11918236 TI - Anaplastic thyroid carcinoma in ecuador: analysis of prognostic factors. AB - The highest incidence of anaplastic thyroid carcinoma (ATC) has been reported in countries with endemic goiter, such as in Ecuador. In this country, ATC is the third most common histologic type of thyroid cancer, following papillary and follicular carcinoma. The aim of this study was to review the clinical presentation and the results of treatment of a large consecutive series of ATC patients treated at the oncological department of a general hospital in Quito, Ecuador. This is a retrospective study of 30 patients diagnosed with ATC at the Social Security Hospital, from 1982 to 1998. Symptomatic rapidly growing neck masses were generally present. All the patients had histological diagnosis of ATC. Two patients with pulmonary metastases and pleural effusion died before treatment could be instituted. Twenty-eight patients received at least one type of treatment: surgery, radiation therapy (RT), or chemotherapy (CT). The two most frequently employed therapeutic modalities were surgery followed by RT and/or CT in 14 patients and surgery alone in 9 patients. Surgery was performed in 23 patients but a complete resection was possible in only 14 patients. RT, postoperatively or alone, was given to 17 patients. Only 5 patients received doses ranging from 4,000 to 5,000 cGy and 4 patients more than 5,000 cGy. CT was administered to 17 patients. Doxorubicin alone was given to 10 patients and different combinations to the remaining patients. Local control was obtained in 8 of 14 complete resections. The prognostic value of the following parameters was studied by univariate analysis: duration of symptoms, size of the tumor, extent of glandular involvement, type of treatment, and surgical margins. A statistically longer survival was found in cases of differentiated carcinoma with areas of ATC or tumor limited to one lobe, those patients who received a complete treatment of chemotherapy, and those patients with tumors smaller than 10 cm and with duration of symptoms longer than 4 months. Longer mean survivals were seen in patients with longer duration of symptoms and smaller lesions. Five patients with focal anaplastic lesions within a differentiated thyroid carcinoma or a lesion limited to one lobe had a significant better survival (a mean of 20 months). PMID- 11918237 TI - Hydatid cyst of the spleen: treatment options. AB - Hydatidosis is a highly endemic parasitic disease caused by Echinococcus. The spleen is infrequently involved in hydatid disease. Splenectomy is the conventional treatment, but there are other treatment options as well. In this article, we reviewed the data of 14 patients with splenic hydatidosis operated on during the last 20 years. The spleen was involved in 2.5% of all abdominal hydatidosis cases during this period and was the only location of disease in 7 of the 14 patients. Partial cystectomy and omentopexy was performed on 2 patients, and splenectomy was performed on 12 patients. In 2 patients, percutaneous drainage was the initial choice of treatment but was not successful, and splenectomy was performed eventually. There was no mortality, and the morbidity rate was 28.6%. Although currently splenectomy is the conventional treatment, experience suggests that partial cystectomy and omentopexy should be the procedure of choice for the treatment of splenic hydatidosis. PMID- 11918238 TI - Three cases of cardiac hydatidosis: diagnosis, surgical treatment, and complications. AB - Three cases of cardiac hydatid disease from among the many cases of hydatidosis (>300) in various organs observed by the authors are reported. The sites of the cysts and the complications that arose are described. The first case developed hydatid pulmonary embolism caused by rupture into the right ventricular cavity, the second suffered peripheral hydatid embolism caused by rupture into the left ventricular cavity, and the third, whose diagnosis was fortuitous, had no complications. The first patient died shortly after admission. The other two underwent radical pericystectomy and partial pericystectomy with cardiopulmonary bypass. The best result was obtained in the third case where rupture had not occurred. The second patient recovered but developed hemiparesis. The various diagnostic tools available are discussed, as well as some technical aspects of pericystectomy, which has a high mortality rate. The importance of early diagnosis and treatment of this rare localization of Echinococcus granulosus is emphasized, and echocardiography is recommended even for nonspecific cardiac symptoms in areas where the parasite is endemic. PMID- 11918239 TI - Gastric outlet obstruction due to duodenal tuberculosis: a case report. AB - A 17-year-old boy was operated on for gastric outlet obstruction; laparotomy revealed a mass with nodules in the first part of the duodenum and multiple mesenteric lymph nodes. The histopathological examination of the duodenal mass and the lymph nodes showed caseating tuberculosis. Because of the rarity of duodenal tuberculosis, the case is reported herewith. PMID- 11918240 TI - A severe type of rectus sheath hematoma requiring surgery: report of a case. AB - This report illustrates the case of a 79-year-old woman in whom a severe type of rectus sheath hematoma was found during conservative therapy for myocardial infarction. Although establishing a correct diagnosis and initiating conservative therapy is usually considered the most appropriate management of this clinical entity, the rectus sheath hematoma in this patient required surgical treatment. PMID- 11918241 TI - Fulminant amoebic colitis: a favorable outcome. AB - Fulminant amoebic colitis (FAC) is often associated with a poor outcome (reported mortality between 55% and 87.5%). We present the outcome of a conservative surgical approach to the management of this condition. Over an 8-year period (January 1992 to December 1999), 23 patients with FAC were managed and prospectively evaluated by a surgical unit at King Edward VIII Hospital, Durban. There were 19 males and 4 females with a mean age of 36 years (range, 18-57 years). Unrelenting abdominal signs was the indication for surgery in all patients. A preoperative diagnosis of FAC was made in 20 patients; in 1 patient the diagnosis was made intraoperatively. FAC was not recognized in 2 patients. The 21 patients diagnosed with FAC were managed by perileal antegrade colonic lavage; 2 patients underwent total colectomy and ileostomy. Following colonic lavage and ileostomy, there was a 95% survival rate (N = 20). Both patients who underwent total colectomy and ileostomy died postoperatively. Patients underwent restorative surgery at a mean period of 7.2 weeks (range, 6-10 weeks) following the initial surgery. The extent of colonic stricturing invariably warranted colonic resection. This included total colectomy (N = 12), right hemicolectomy (N = 5), left hemicolectomy (N = 2), and right and transverse colectomy (N = 1). The overall survival rate of the 23 patients presenting with FAC was 82.6% (N = 19). Early recognition of FAC and a conservative surgical approach are associated with a favorable outcome. Resectional surgery in FAC has a potentially fatal outcome and should be avoided. PMID- 11918242 TI - The management of penetrating neck injuries. AB - In recent years, there has been a major increase in patients with penetrating injuries to the neck admitted to the Johannesburg Hospital. Pressure on resources led to increasing delays for surgery, and a policy of selective conservatism emerged. In common with other centers, mandatory exploration of all wounds that breach the platysma was found to be no longer necessary as it became clear that many penetrating wounds to the neck were best treated conservatively. A policy of blanket investigation of all nonoperated cases also matured toward selective investigation, directed by careful clinical examination. A retrospective study was made of all patients undergoing exploration for gunshot wounds or stabs to the neck at the Johannesburg Hospital Trauma Unit between 1994 and 1998. An overall mortality rate of 9% was mostly a reflection of severe, associated injuries. The evolution of the nonoperative management of cervical penetrating wounds is a good example of the validity of the concept of "selective conservatism." A distillation of the experience at a busy, urban trauma center is presented, with guidelines to manage these potentially lethal injuries. PMID- 11918243 TI - Selective management of penetrating neck injuries based on clinical presentations is safe and practical. AB - Selective management was offered in 57 patients who had penetrating injury to the anterior neck. During the study period, decision making in patient management at our institution depended largely on clinical presentations. Indications for neck explorations were unstable hemodynamics, airway obstruction, active bleeding from the wound, and evidence of aerodigestive tract injuries. Some patients with deep wounds of zone II also underwent neck explorations. Investigations were performed in selected cases. With this selective policy, there were two unnecessary operations among 40 patients (70.2%) who underwent neck exploration. Both of them were operated because of deep wounds of zone II. The remaining 17 patients (29.8%) had uneventful conservative treatment. There was no mortality in this study. The authors concluded that selective management of penetrating neck injuries based on clinical presentations is safe and practical. PMID- 11918244 TI - Is age a risk factor for esophagectomy? AB - In recent years, the number of elderly patients with esophageal cancer as well as the number submitted for esophageal resection has been increasing. With respect to nutritional and pulmonary evaluations, surgical staging, and mortality, 37 patients over the age of 65 who underwent esophagectomy and simultaneous reconstruction were analyzed. This group was compared statistically with a group of 162 patients younger than 65 to determine whether age was a factor influencing treatment and outcome. There was no statistical difference between the groups relating to the described variables. Age should not be a limiting factor when it comes to offering an aggressive surgical approach for the esophageal cancer patient aged 65 or more. This approach can be performed as safely in older patients as it is in younger patients, with similar incidence of mortality. PMID- 11918245 TI - Complicated choledochal cysts. AB - Choledochal cysts have a potential for causing a variety of complications. This article presents the incidence, types, and presentation of complications in patients with choledochal cysts and the results of surgery in these patients. Prospectively collected data of 81 patients with choledochal cysts treated at a tertiary care center were retrospectively analyzed. Sixteen (20% of surgical patients; 100% of study group) patients were found to have a complicated choledochal cyst that either manifested as an atypical presentation or necessitated a change in the treatment planning. Eight (50%) of these patients had complications of infective nature, and another 8 (50%) had noninfective complications. Fifteen (94%) patients with complications were adults and 11 (63%) patients had Todani type-IVa cysts; a cystic type of intrahepatic component in 10 patients. Five (31%) patients had undergone prior cystenteric drainage operations. Excision of the choledochal cyst was possible in 14 (88%) patients under both emergency and elective situations; cyst excision was performed in the second stage after an initial external drainage procedure in two of these patients. There was no mortality and no major postoperative complications. Complications of choledochal cysts are more likely in adult patients and in those with both intra- and extrahepatic cysts (Todani type-IVa). Despite the presence of complications, the choledochal cyst can be safely excised in a majority of patients. PMID- 11918246 TI - Intracavernosal alprostadil is effective for the treatment of erectile dysfunction in diabetic men. AB - The efficacy and safety of intracavernosal alprostadil was evaluated for the treatment of erectile dysfunction in men with type I or type II diabetes mellitus. This was an open-label, flexible dose-escalating study involving 336 men (77% of whom were Asian/Oriental) enrolled by 15 centres in Australia, Canada and seven countries in Asia. The effective alprostadil dose, ie the dose producing penile rigidity adequate for intercourse and lasting up to 60 min, was established by titration at the clinic prior to entry into the 6 month self treatment home phase. All men were fully trained in the self-injection technique before entry into the home phase. Efficacy and safety were assessed using patient and partner diaries and by interview at clinic visits during the titration phase and after 1, 3 and 6 months of treatment. An effective home dose was established by titration for 94% of the 336 men (median dose 20 microg, range 2.5-60 microg). Of 278 (83%) men who entered the home phase, 277 men (247 with type II diabetes and 30 with type I diabetes) had evaluable data for alprostadil dosage and clinical response. During the home phase, a satisfactory erectile response was achieved after 99% of injections, and the median alprostadil dose remained unchanged. The initial home dose and clinical response were similar in type I and type II diabetic men. Treatment was generally well tolerated with a low incidence of penile pain (24%) In conclusion, intracavernosal alprostadil was effective and well tolerated in type I and type II diabetic men with erectile dysfunction of mixed aetiology. PMID- 11918247 TI - An investigation into the relationship between prostate size, peak urinary flow rate and male erectile dysfunction. AB - This study sought to identify whether a true relationship exists between benign prostatatic hyperplasia (BPH) and erectile dysfunction (ED). In a community-based study, 427 men underwent transrectal ultrasound (TRUS), uroflow studies and a questionnaire concerning erectile function. ED had a significant correlation to age (r = 0.19, P < 0.001). But comparisons of prostate volume and analysis of maximum flow rate showed no significant difference between three erectile functional groups; ranging from no ED to complete ED, (one way analysis of variance). However when these two parameters were correlated to age a significant association was found to exist (log prostate volume; r= 0.26, P < 0.001, log maximum flow rate; r= -0.13, P= 0.02). Prostate size and uroflow studies show no correlation with ED, but ED and BPH had a significant correlation with ageing. This makes a direct association between male ED and BPH unlikely but supports the theory that the association between the two pathologies could be due instead to the common link of ageing. PMID- 11918248 TI - Penile prosthesis infections. AB - In this retrospective review we looked at infection rates and risk factors in penile prosthesis recipients. Computer registry records of patients implanted with three-piece inflatable prostheses from April 1986 to March 1999 were reviewed. Bacteriological culture report data were obtained from the medical records. There were 491 three-piece inflatable penile prosthesis recipients with follow-up ranging from 1 to 168 months (mean 83). All patients were implanted more than 1 y ago. There were 10 infections in 491 recipients (2.0%). In 285 primary prosthesis recipients there were seven infections (2.5%). In 206 secondary prosthesis recipients there were three infections (1.5%). Seven infections occurred in 354 non-diabetics (2.0%) and three infections occurred in 137 diabetics (2.2%). None of these differences were statistically significant. Explants for infection were performed in 8 weeks or less following implantation in eight patients. In two patients explants occurred at 17 and 26 months following implantation. Cultures from the periprosthetic space grew E. coli (four), Proteus mirabilis (two), Staph. aureus (two), and bacteroides (two). Infections occurred in 2.0% of three-piece inflatable penile prosthesis recipients. There were no statistically significant differences in infection rates between first time and repeat prosthesis recipients and between diabetic and non-diabetic recipients. Eight of the 10 infections occurred 8 weeks or less following implantation. PMID- 11918249 TI - Does vibration offer any advantage over visual stimulation studies (VSS) in the assessment of erectile capacity? AB - The purpose of this work was to determine whether vibration stimulation or erotic videotape material can provide an acceptable diagnostic yield for patients with erectile dysfunction (ED) without the patient needing to endure more explicit erotic films, which may be distasteful to some patients, or intracavernous injections, to which there may be a high inhibitory response. Ninety-five subjects were randomly exposed to either vibration or videotape alone and erectile response monitored by the RigiScan. Where no clinical response was recorded by the RigiScan or by self-report by the patient, both stimuli were presented to the subject. Forty-nine subjects received vibration first and 46 received videotape first. Use of clinically based standardised measures revealed neither group achieving above threshold responses to the first stimulation and there were no differences between the two groups. Eight subjects (8%) between both groups exceeded the tip threshold during combination stimulation. Single stimulation with vibration or erotic videotape with provocative RigiScan monitoring is unhelpful in the assessment of ED in any of the diagnostic subgroups of ED. In comparison to previous provocative studies, combination of stimuli sets, whilst increasing penile response (circumferential change and rigidity) did not lead to significant evidence of clinically relevant responses using current RigiScan measures or patient self-report of change. Further studies are necessary to determine the most useful set of stimuli for provocation studies with the RigiScan. PMID- 11918250 TI - Erectile dysfunction in a primary care setting: results of an observational, no control-group, prospective study with sildenafil under routine conditions of use. AB - In order to assess the effectiveness of sildenafil under routine conditions of use in primary care settings and to evaluate its impact on patient's life satisfaction and partner's satisfaction with treatment for erectile dysfunction (ED), an open, multicentre, observational, prospective study was designed in which 2816 patients were treated with sildenafil for at least 10 weeks. Effectiveness was assessed using the International Index of Erectile Dysfunction (IIEF), life satisfaction was measured with 'Life-satisfaction Check List' (LISAT 8), and EDITS was optionally used to assess the partner's satisfaction with ED therapy. Sildenafil was effective in 86.6% of patients. All dimensions of IIEF significantly increased with sildenafil, particularly erectile domain which overall sample mean score improved was 13.2 points (P < 0.001). The greatest increases in satisfaction with all aspects of life were seen in sex life and relationship with partner dimensions. The patients' partners, answered by a minority of partners, were highly satisfied with the treatment and its rapid action, therefore they were in favour of continuing with same. The adverse events occurring were similar to those seen in clinical research on sildenafil in the premarketing phase. No control group was included in this study. PMID- 11918252 TI - Herpes zoster producing temporary erectile dysfunction. AB - Varicella Zoster affecting the sacral dermatomes is a rare but well recognised cause of urinary retention. Only one case of erectile dysfunction associated with Varicella Zoster has previously been described, which was longstanding, but no cases of transient erectile dysfunction following Zoster infection are recorded. We present one such case. PMID- 11918251 TI - Effects of beta-blockers on sexual performance in men with coronary heart disease. A prospective, randomized and double blinded study. AB - In a prospective trial assessing the effects of beta-blockers on sexual function men with coronary heart disease were randomized to a 4 month treatment with sustained release metoprolol 95 mg or placebo. A standardized and validated self report questionnaire (KEED = Kolner Erhebungsbogen der Erektilen Dysfunktion) dealing with several aspects of sexual performance in men had to be answered at the beginning and at the end of the study. Based on 65 patients completing the study, sex life seemed unaffected by metoprolol treatment. PMID- 11918253 TI - Highly selective embolization of bilateral cavernous arteries for post-traumatic penile arterial priapism. AB - High-flow priapism is characteristically diagnosed on clinical findings: a prolonged, non-painful erection with a delayed onset that develops after a penile or perineal trauma. If conservative measures fail arteriography is indicated, which shows a blush of extravasating contrast from an arterio-cavernous fistula (rarely, as in our case bilateral) that can be treated by embolization. The embolic agent is gelfoam or a microcoil. Bilateral embolization is indicated when unilateral treatment does not result in detumescence of the penis. When the embolization is done highly selective the risk of complications is low and the results on erectile function are good. PMID- 11918254 TI - Heparin-induced priapism. AB - Heparin-induced priapism constitutes a special form of pharmaco-induced prolonged erection, but the pathophysiological principles are not yet definitely clear. Heparin-induced antiplatelet-antibodies may lead to the aggregation of thrombocytes and thus alter the penile blood flow leading to low-flow priapism. Alternatively, this condition may be explained by initial high-flow priapism that later turns into ischemic priapism. The question remains whether hemorrhage with subsequent organisation of the hematoma and late fibrosis constitutes a pathogenetic factor. Besides this pathogenetic discussion, this paper presents the differential diagnosis of priapism as well as diagnostic and therapeutic procedures. PMID- 11918256 TI - Reply to: Is there a role of radial rigidity in the evaluation of erectile dysfunction? By Ku JH, Song YS, Kim ME, Lee NK and Park YH. PMID- 11918257 TI - Veterinarians take a lead role in agency responses to bioterrorism. PMID- 11918258 TI - Nashville disaster management program to zero in on responders. PMID- 11918259 TI - Progress made in feline sarcoma research. PMID- 11918260 TI - Promoting the veterinarian's role in public health. PMID- 11918262 TI - Still more thoughts on ovarian remnant syndrome. PMID- 11918263 TI - Still more thoughts on ovarian remnant syndrome. PMID- 11918264 TI - Rodeos--from a bull's point of view. PMID- 11918265 TI - Appreciative of helpful comments on urine marking in cats. PMID- 11918266 TI - Analyzing the cost of educational debt. PMID- 11918267 TI - What is your diagnosis? Avascular necrosis of the left femoral head with secondary osteoarthritis of the hip joint. PMID- 11918268 TI - Fringe benefits received by veterinarians, 1999. PMID- 11918270 TI - Surgical correction of double-chambered right ventricle in dogs. AB - Double-chambered right ventricle (DCRV) is possibly an emerging congenital cardiac anomaly in dogs. The defect causes clinical and pathophysiologic signs similar to those of congenital pulmonic stenosis in dogs but has distinct diagnostic features, breed predilections, and implications for treatment. The defect is often associated with clinical signs early in life. Surgical correction of DCRV can be undertaken with the aid of cardiopulmonary bypass and offers the prospect of an improved clinical outcome. PMID- 11918269 TI - Isolation of shiga-toxigenic Escherichia coli O157 from hide surfaces and the oral cavity of finished beef feedlot cattle. AB - OBJECTIVE: To determine whether viable shiga-toxigenic Escherichia coli (STEC) O157 could be isolated from hide surface locations and the oral cavity of finished beef feedlot cattle. DESIGN: Within-animal prevalence distribution survey. ANIMALS: 139 finished cattle in 4 pens in a feedlot in Nebraska; prevalence of fecal STEC O157 shedding ranged from 20 to > 90%. PROCEDURE: Samples were collected from 7 sites from each animal: feces, oral cavity, and 5 hide surface locations (lumbar region, ventral aspect of the neck, ventral abdominal midline [ventrum], dorsal thoracic midline [back], and distal aspect of the left hind limb [hock]). RESULTS: Viable STEC O157 were isolated from the oral cavity or 1 or more hide surfaces of 130 cattle, including 50 fecal isolation negative cattle. Site-specific prevalence of STEC O157 was 74.8% for oral cavity samples, 73.4% for back samples, 62.6% for neck samples, 60.4% for fecal samples, 54.0% for flank samples, 51.1% for ventrum samples, and 41.0% for hock samples. Only 5 cattle tested negative for STEC O157 at all 7 sites. Multiple correspondence and cluster analyses demonstrated that bacterial culture of feces, oral cavity samples, and back samples detected most cattle with STEC O157. CONCLUSIONS AND CLINICAL RELEVANCE: Results suggest that viable STEC O157 may be isolated from the oral cavity, multiple hide surfaces, and feces of a high percentage of fed beef cattle and that bacterial culture of feces alone generally underestimates the percentage of fed beef cattle from which STEC O157 can be isolated. PMID- 11918271 TI - Results of cerebrospinal fluid analysis, neurologic examination findings, and age at the onset of seizures as predictors for results of magnetic resonance imaging of the brain in dogs examined because of seizures: 115 cases (1992-2000). AB - OBJECTIVE: To determine whether neurologic examination findings, results of CSF analysis, or age at the onset of seizures could be used to predict whether results of magnetic resonance imaging (MRI) would be normal or abnormal in dogs with seizures. DESIGN: Retrospective study. ANIMALS: 115 dogs. PROCEDURE: Information on results of neurologic examination, results of CSF analysis, age at the onset of seizures, and results of MRI was obtained from the medical records. RESULTS: Results of MRI were abnormal in 61 dogs and normal in 54. Sensitivity and specificity of neurologic examination alone were 77 (47/61) and 91% (49/54), respectively. Sensitivity and specificity of CSF analysis alone were 79 (48/61) and 69% (37/54), respectively. Results of MRI were abnormal for 12 of 28 (43%) dogs with abnormal CSF analysis results and normal neurologic examination results but for only 2 of 35 (6%) dogs with normal CSF analysis and normal neurologic examination results. Similarly, results of MRI were abnormal for 36 of 37 (97%) dogs with abnormal CSF analysis and abnormal neurologic examination results but for only 11 of 15 (73%) dogs with normal CSF analysis results and abnormal neurologic examination results. Age at the onset of seizures (< 6 vs > or = 6 years old) was not significantly associated with results of MRI. CONCLUSIONS AND CLINICAL RELEVANCE: Results suggest that neurologic examination findings and results of CSF analysis are useful in predicting whether results of MRI will be abnormal in dogs examined because of seizures, but age at the onset of seizures is not. PMID- 11918272 TI - Risk factors for Clostridium piliforme infection in foals. AB - OBJECTIVE: To determine risk factors for Clostridium piliforme infection in neonatal foals on a Thoroughbred breeding farm in California. DESIGN: Case control and retrospective cohort studies. ANIMALS: 322 neonatal Thoroughbred foals either born on the study farm or born elsewhere but traveled to the farm with their dam during the 1998, 1999, and 2000 breeding seasons. PROCEDURE: Mare and foal records from 1998, 1999, and 2000 were examined, using case-control design methods to determine variables associated with increased risk of C. piliforme infection in foals. Important risk factors identified in the case control study were then reevaluated by use of a retrospective cohort design, using data from all neonatal foals present on the farm during the 3-year study period. RESULTS: Foals born between March 13 and April 13 were 7.2 times as likely to develop C. piliforme infection as were those born at any other time of the foaling season. Foals of nonresident (visiting) mares were 3.4 times as likely to develop disease as were foals born to mares that were permanent residents of the study farm. Foals of mares < 6 years of age were 2.9 times as likely to develop disease as were foals born to older mares. CONCLUSIONS AND CLINICAL RELEVANCE: Results of this research can be used to better understand the epidemiologic factors of C. piliforme infection in horses. High-risk foals can be closely monitored to aid in early diagnosis and treatment, resulting in the best possible clinical outcome for affected individuals. PMID- 11918273 TI - Evaluation of a commercially available immunoassay for assessing adequacy of passive transfer in calves. AB - OBJECTIVE: To evaluate diagnostic utility of a commercially available immunoassay for assessing adequacy of passive transfer of immunity in neonatal calves. DESIGN: Prospective study. ANIMALS: 123 calves. PROCEDURE: Blood and serum samples were obtained from the calves prior to 2 weeks of age. The immunoassay was performed, along with refractometry and an 18% sodium sulfite turbidity test. Serum IgG concentration was determined with a radial immunodiffusion assay. Sensitivity and specificity of the immunoassay, refractometry, and the sodium sulfite test were calculated by comparing results with results of the radial immunodiffusion assay. RESULTS: Sensitivity and specificity of the blood IgG immunoassay were 0.93 and 0.88, respectively, compared with 1.00 and 0.53 for the sodium sulfite test. For refractometry, sensitivity and specificity were 0.71 and 0.83, respectively, when a serum total solids concentration of 5.2 g/dl was used as the cutoff between positive and negative test results. CONCLUSIONS AND CLINICAL RELEVANCE: Results suggest that the immunoassay performs well in detecting calves with inadequate passive transfer of immunity. PMID- 11918274 TI - Microbiologic evaluation of commercial probiotics. AB - OBJECTIVE: To evaluate contents of commercial probiotic products marketed for veterinary or human administration. DESIGN: Microbiologic culture assay. SAMPLE POPULATION: 8 veterinary probiotics and 5 human probiotics. PROCEDURE: Quantitative bacteriologic culture was performed on all products, and isolates were identified via biochemical characteristics. Comparison of actual contents versus label claims was performed. RESULTS: Label descriptions of organisms and concentrations accurately described the actual contents of only 2 of 13 products. Five veterinary products did not specifically list their contents. Most products contained low concentrations of viable organisms. Five products did not contain 1 or more of the stated organisms, and 3 products contained additional species. Some products contained organisms with no reported probiotic effects; some of these organisms could be pathogens. CONCLUSIONS AND CLINICAL RELEVANCE: Most commercial veterinary probiotic preparations are not accurately represented by label claims. Quality control appears to be poor for commercial veterinary probiotics. PMID- 11918275 TI - Primary biliary cirrhosis: lessons learned from an organ-specific disease. AB - Primary biliary cirrhosis is an autoimmune liver disease that predominantly affects women and is characterized by chronic progressive destruction of small intrahepatic bile ducts with portal inflammation and subsequent fibrosis. The serological hallmark is the presence of antimitochondrial antibodies, which are found in 95% of patients. These antibodies are directed against the 2-oxo-acid dehydrogenase complexes located on the inner membrane of mitochondria. Although the role of antimitochondrial antibodies in the pathogenesis is unknown, the presence of antibodies has allowed detailed immunological definition of the antigenic epitopes, the autoantibodies, and the T-cell response. Theories have been proposed regarding the mechanism of immune-mediated bile duct damage in primary biliary cirrhosis, including the possible role of T-cell-mediated cytotoxicity and molecular mimicry. Primary biliary cirrhosis is usually diagnosed based on the triad of elevated alkaline phosphatase, antimitochondrial antibodies, and characteristic histological changes on liver biopsy. Biochemical liver abnormalities are consistent with the presence of cholestasis and include an elevation of both serum alkaline phosphatase and gamma-glutamyl transpeptidase, with or without elevation of aminotransferase levels. Ursodeoxycholic acid, a dihydroxy bile acid, appears to be the only effective therapy in preventing or delaying the need for liver transplantation. However, a number of patients receiving ursodeoxycholic acid still develop progressive disease and require transplantation; at present, liver transplantation is the only effective therapy for end-stage primary biliary cirrhosis. PMID- 11918276 TI - Glucocorticoid receptors in lymphocytes and stability of kidney graft function. AB - The glucocorticoid receptors in lymphocytes of patients treated with glucocorticoids after kidney transplantation have been studied in order to determine whether abnormalities in corticosteroid binding and trans-activation of steroid-receptor complexes, i.e., their translocation into nuclei, may contribute to the resistance of patients to glucocorticoid therapy. The patients were divided into two groups, according to graft stability: patients with stable graft function and those with chronic allograft rejection. The study revealed changes in both level and binding affinity of glucocorticoid receptors in peripheral blood lymphocytes from patients with chronic graft rejection, compared with control level, as well as with values of patients with stable graft function. These data indicate that sensitivity to glucocorticoids depends, at least in part, on the alterations of glucocorticoid receptors. The receptor translocation into nuclei indicates that unknown post-receptor events might also be involved in glucocorticoid resistance that seriously impair successive glucocorticoid therapy after organ transplantation. Further examination of glucocorticoid receptors in cases of organ transplantation seems warranted. PMID- 11918277 TI - Erythrocyte membrane and cytoskeletal protein glycation and oxidation in short term diabetic rabbits. AB - The objective of this study was to elucidate the glycation and oxidation processes in plasma and erythrocyte membrane proteins as well as the major erythrocyte cytoskeletal protein, spectrin, using a short-term experimental rabbit diabetes model. Diabetes was induced with a single-dose alloxan injection. Spectrin was purified from erythrocyte ghosts with selective solubilization followed by gel filtration chromatography techniques, and tested for purity using sodium dodecyl sulfate-poly-acrylamide gel electrophoresis. Glycation in plasma proteins was measured as fructosamine using the nitroblue tetrazolium method, and in erythrocyte membrane and purified spectrin, as ketoamine equivalents, by the hydrazine/phenylhydrazine method. Protein oxidation in plasma, erythrocyte membrane proteins, and purified spectrin was evaluated in terms of sulfhydryl oxidation, based on cis-dichlorodiammine platinum (II) binding. Carbonyl formation was also measured in plasma and membrane proteins. Sulfhydryl oxidation, carbonyl groups and glycated protein levels showed statistically significant differences between the diabetic and control groups for both the plasma and the erythrocyte membrane proteins. The cis-dichlorodiammine platinum (II) binding was significantly different in diabetic rabbit erythrocyte spectrin, while glycation was not significantly different for this protein. Our data clearly demonstrate that both protein glycation and oxidation are biochemical alterations occurring in diabetes, even of short duration. PMID- 11918278 TI - Antioxidant enzymes and paraoxonase show a co-activity in preserving low-density lipoprotein from oxidation. AB - Oxidative modification of low-density lipoprotein in the artery wall plays a crucial role in the development of atherosclerosis. This physiopathological mechanism is clearly inhibited by high-density lipoprotein possibly via paraoxonase enzyme activity, present in high-density lipoprotein. In this study, we determined the in vitro susceptibility of low-density lipoprotein to oxidation and the effect of various factors, such as paraoxonase phenotypes, on this process. Low-density lipoprotein from healthy volunteers (n=66) was isolated using the precipitant reagent and the oxidation was evaluated by measuring the malonyl dialdehyde and diene levels. Low-density lipoprotein cholesterol and phospholipid, vitamin E, serum cholesterol, high-density lipoprotein and low density lipoprotein cholesterol levels, and erythrocyte antioxidant enzymes were also determined. There was no difference among the parameters with regard to gender. Low-density lipoprotein samples obtained from subjects with the AA allele were more prone to oxidation, as observed by their higher stimulated conjugated diene (P=0.041) and thiobarbituric acid-related substance (P=0.042) levels, than samples from subjects with AB or BB alleles. The subjects with the BB allele had higher superoxide dismutase (P=0.021) and catalase (insignificant increase) activities, while their conjugated diene (P=0.000) levels were lower. In conclusion, our results revealed that the high low-density lipoprotein oxidation is related to the high low-density lipoprotein cholesterol content and low phospholipid content. The present study demonstrated an increase in superoxide dismutase and catalase activities, as well as PON1 activities, in subjects with the BB allele. Since these enzymes all show activity against low-density lipoprotein oxidation, we propose that future investigations on atherosclerotic processes should address PON1 polymorphism as well as PON1 and other antioxidant enzymes. PMID- 11918279 TI - The determination of ketone bodies: preanalytical, analytical and physiological considerations. AB - The arterial ketone body ratio is calculated as the ratio of arterial levels of acetoacetate/beta-hydroxybutyrate. It correlates with survival in experimental hemorrhagic shock and outcome after liver surgery and myocardial infarction. Procedures for determination of ketone bodies are often laborious and unreliable. As yet the relationship between results from arterial and venous samples is unclear. We therefore describe the determination of the ketone bodies acetoacetate and 3-hydroxybutyrate by an easy, reliable, rapid, inexpensive enzymatic assay using 3-hydroxybutyrate dehydrogenase (E.C. 1.1.1.30) in a semi automated setting that does not require deproteinization. Preanalytical parameters, including separation from corpuscular elements within 1 h and storage on ice for less than 1 h, must be strictly observed to avoid rapid decay of acetoacetate by spontaneous decarboxylation. The assay has high sensitivity, specificity (+/-5%), and precision (CV <2.5%) with a measurable range of 5-500 micromol/l for either ketone body, and requires only 23.5 microl of plasma. At temperatures below -17 degrees C plasma may be stored for prolonged periods. Results from prospectively scheduled simultaneous sampling of arterial blood and venous blood from the right atrium in 100 consecutive patients with severe multiple trauma (mean Injury Severity Score 38+/-13) support the view that the lung has no role in ketone body metabolism. We conclude that central venous blood can safely be substituted for arterial blood for determination of the ketone body ratio. PMID- 11918281 TI - D-dimer increase after percutaneous transluminal angioplasty and clinical recurrence after primary revascularization in acute myocardial infarction? A pilot study. AB - It has been reported that the increase of plasminogen activator inhibitor-1 activity immediately after elective coronary angioplasty is related to subsequent clinical recurrence in patients with chronic coronary artery disease. The aims of our study were to evaluate the behavior of plasminogen activator inhibitor-1 and D-Dimer after revascularization in acute myocardial infarction patients treated with angioplasty and stenting and if this behavior is predictive of subsequent clinical recurrence. D-Dimer and plasminogen activator inhibitor-1 activity were evaluated in two groups of patients. Group 1 consisted of 54 consecutive patients undergoing primary angioplasty for acute myocardial infarction and Group 2 consisted of 48 patients undergoing elective angioplasty. Patients underwent control coronary angiography only in the case of clinical recurrence and/or positivity of provocative tests. D-Dimer and plasminogen activator inhibitor-1 baseline levels were significantly higher in group 1 than in group 2 (P<0.0005 and P<0.05, respectively). The percentage of group 1 patients with a post procedural increase in D-Dimer was significantly higher among those with subsequent clinical recurrence with restenosis (61%) than among those with no recurrence (25%, P<0.05). No difference was observed in group 2. The percentage of group 2 patients in whom no decrease of plasminogen activator inhibitor-1 was observed after angioplasty was significantly higher (83%) among those with subsequent recurrence than among those with no recurrence (38%, P<0.05). This pattern was not observed in group 1. In conclusion, the role of early changes in plasminogen activator inhibitor-1 in predicting clinical recurrence after primary angioplasty in acute myocardial infarction patients is less clear than that observed after elective angioplasty. A significant role seems to be played by a more-marked clotting activation with increased fibrin formation. PMID- 11918280 TI - Homocysteine and atheromatous renal artery stenosis. AB - Hyperhomocysteinemia is an independent risk factor for vascular disease, frequently observed in patients with severe renal impairment. Hyperhomocysteinemia has never been considered as a possible risk factor in renal artery stenosis. We investigated plasma folate and vitamin B12, methylenetetrahydrofolate reductase (MTHFR) C677T and cystathionine beta-synthase (CBS) 844ins68 polymorphisms, and homocysteine levels before and after methionine (100 mg/kg) loading in 58 patients with angiographically documented renal artery stenosis and mildly impaired renal function. One hundred and two normotensive subjects with angiographically normal coronary arteries and no history or clinical or angiographic evidence of atherosclerosis in other vascular districts, were considered as a control group. Mean total homocysteine levels were significantly higher in patients than in controls (P<0.01), as was the prevalence of hyperhomocysteinemia (51.7% vs. 32.3%, P<0.05). However, MTHFR alleles and genotypes as well as CBS 844ins68 mutation frequencies were similar in the two groups, whereas a lower folate level was observed in the patients. Moreover, patients with MTHFR A/A genotype showed a poorer folate status than control subjects, suggesting that a subclinical folate deficiency may be very frequent in renal artery stenosis, regardless of C677T mutation. In conclusions, hyperhomocysteinemia is common in patients with atheromatous renal artery stenosis; a subclinical folate deficiency seems to be involved, regardless of MTHFR thermolabile or CBS insertion genotypes. Folate supplementation might be useful in the management of overall vascular risk of these patients. PMID- 11918282 TI - Targeting the "species gene ensemble". AB - The members of a given species display a normal distribution of gene expression which is here termed the species gene ensemble. This more specific designation of the 'genetic background' leads to a consideration of why gene targeting outcomes are regulated by the species gene ensemble. One consequence is the utility of the heterozygous knockout which buffers the drastic compensatory reactions of the homozygous knockout thereby revealing subtle but yet consistent alterations in hippocampally-dependent behaviors. PMID- 11918283 TI - Behavioral analysis of CREB alphadelta mutation on a B6/129 F1 hybrid background. AB - The cyclic AMP (cAMP)-response element binding protein (CREB) is an activity dependent transcription factor that plays a role in synaptic plasticity and memory storage in Aplysia, Drosophila, and rodents. Mice with targeted deletions of two CREB isoforms (alpha and delta; CREB alphadelta mice) have been characterized on a mixed genetic background of C57BL/6 (B6) and 129/SvEv (129), as well as on a defined F1 hybrid of B6 and FVB/N, and these results suggest that the phenotype of CREB alphadelta mice depends critically on genetic background. In an examination of the hypothesis that the role of CREB in learning and memory can be influenced by strain differences, we analyzed mice with the CREB alphadelta mutation on an F1 hybrid background of B6 and 129 strains. CREB alphadelta mice on this background had impaired short-term and long-term cued and contextual fear conditioning and normal spatial learning in the Morris water maze. Our results suggest that at least some aspects of hippocampal function are normal in CREB alphadelta mice, and that CREB alphadelta mice on the B6/129 F1 background have alterations in amygdala function. These studies underscore the importance of controlling for genetic background in the behavioral analysis of knockout and transgenic mice. PMID- 11918284 TI - Genetic analysis and the targeting of hippocampal function. PMID- 11918285 TI - Long-term monitoring of hippocampus-dependent behavior in naturalistic settings: mutant mice lacking neurotrophin receptor TrkB in the forebrain show spatial learning but impaired behavioral flexibility. AB - Previous behavioral studies (Minichiello et al., Neuron 1999;24:401-414) showed that mice deficient for the TrkB receptor in the forebrain were unable to learn a swimming navigation task with an invisible platform and were severely impaired in finding a visible platform in the same setup. Likewise, additional behavioral deficits suggested a malfunction of the hippocampus and proximally connected forebrain structures. In order to discriminate whether the behavioral impairment was caused either by deficits in spatial memory and learning, or alternatively by loss of behavioral flexibility, 8 trkB mutant, 13 wild-type, and 22 heterozygous mice were implanted with transponders and released for 21 days into a large outdoor pen (10 x 10 m). The enclosure contained 2 shelters and 8 computer controlled feeder boxes, delivering food portions for every mouse only during their first visit. Every third day, mice received food ad libitum inside the shelters. All mice learned to patrol the boxes correctly within a few days. However, significant differences emerged during those days with free food available. Wild-type mice remained inside the shelters, while all homozygous mutants continued to patrol the boxes in their habitual way, the heterozygous mutants showing intermediate scores. These and previous data suggest that one of the natural functions of the mouse hippocampus is to comediate behavioral flexibility, and that TrkB receptors might play an essential role in maintaining the neuronal short-term plasticity necessary for this capacity. PMID- 11918286 TI - Behavioral and neuroanatomical characterization of the Fmr1 knockout mouse. AB - Previous studies showed the Fmr1 knockout (KO) mouse to be an excellent animal model for human fragile-X syndrome. The aim of this study was to further characterize the phenotype of these animals. Neuroanatomically, KO male mice were compared to wild-types (littermates) with respect to their sizes of hippocampal intra- and infrapyramidal mossy fiber (IIPMF) terminal fields. Behaviorally, they were tested in four different paradigms, each measuring different aspects of cognitive and emotional behavior: elevated plus maze (anxiety), neutral cage (aggression), open field (exploration), and radial maze (spatial memory). The results showed a diminished ability for radial maze learning associated with smaller sizes of IIPMF terminal fields. In addition, Fmr1 knockout animals exhibited increased locomotor activity, while no differences were found for aggression and anxiety. These data suggest the involvement of FMRP protein in the development of spatial learning and the sprouting of IIPMF terminal fields. PMID- 11918287 TI - Force transducer-based movement detection in fear conditioning in mice: a comparative analysis. AB - Fear conditioning (FC) allows the dissociation of hippocampal and nonhippocampal behavioral function in rodents, and has become a diagnostic tool in transgenic mouse research employed to investigate mutation-induced changes in brain function. Although the procedural details of the paradigm have been established, quantification of the behavioral output, freezing, remains problematic in mice. Observation-based techniques are time-consuming and may be subject to bias, while movement detection with photocells is imprecise. Here we describe an alternative method for movement detection, based on an electronic force transducer system that allows the quantification of acceleration forces generated by a moving subject. We compare the behavior of two inbred strains of mice (C57BL/6 and DBA/2) whose performance is known to differ in hippocampal tasks, including FC. The comparison is made using multiple techniques: the force transducer approach, and three observation-based methods, a computer-aided event-recording approach, a traditional time-sampling paper/pencil method, and a subjective impression-based scoring system. In addition, we investigate the correlation structures of behavioral elements quantified by event recording, using principal component analyses; we conclude that fear may manifest in multiple forms and in a stimulus- and genotype-dependent manner. We suggest that the force transducer system provides precise quantification of movements in an automated manner and will allow high-throughput screening for mutation and drug effects in mice. However, we also argue that fear responses can be complex, and freezing behavior may not be the only measure of fear or fear-associated memory. PMID- 11918288 TI - Altered mossy fiber distributions in adult Fmr1 (FVB) knockout mice. AB - The fragile-X mental retardation protein (FMRP) is greatly reduced or absent in individuals with fragile-X mental retardation syndrome, a common, heritable form of mental retardation. Morphological studies suggest that this protein functions in normal synapse maturation and neuronal plasticity. Examination of human brain autopsy tissue has shown that fragile-X patients exhibit long, thin spines more frequently, and stubby mushroom-shaped spines less frequently, than these two types of spines are seen in normal autopsy tissue. Fragile-X tissue also has a greater density of these spines along dendrites, which suggests a possible failure of synapse elimination. Fmr1 knockout mice and wild-type littermates brains were processed for Timm staining, which reveals the zinc-rich terminals of the dentate gyrus, the mossy fibers. The Fmr1 knockout mice exhibited a pattern of Timm granule-staining within the stratum oriens of subfield CA3 and the inner molecular layer that was significantly different than staining seen in wild-type animals. The sources and consequences of the altered terminal staining are unclear, but are discussed in relation to immature synapse morphology, a failure of normal regression of synapses, and a potential biological penalty of such a failure to regress. PMID- 11918289 TI - Age-dependent differential regulation of genes encoding APP and alpha-synuclein in hippocampal synaptic plasticity. AB - We investigated the modulation of the messenger RNA encoding the amyloid precursor protein (APP) and alpha-synuclein following induction of long-term potentiation (LTP) in the dentate gyrus of young and aged rats. Three hours after tetanic stimulation, LTP induced in the young rats was maintained; the aged rats, however, fell into two subgroups: those in which LTP was maintained, and those in which LTP had declined to basal levels. In young rats, the global expression of mRNAs of all isoforms of APP and in particular that of the isoform lacking the KPI domain were significantly upregulated. In aged rats, the global expression of mRNAs of all isoforms of APP was not modified, regardless of whether LTP was maintained or not. The level of mRNA encoding the Kunitz protease-inhibitory (KPI)-minus isoform of APP, however, was increased in aged rats in which LTP was maintained, suggesting that the gene of this isoform may be more specifically regulated by synaptic plasticity. In contrast, we found that the gene encoding alpha-synuclein showed a trend towards being downregulated at the mRNA level in young rats following LTP, and significantly so in aged rats in which LTP was maintained, whereas it was not downregulated in aged rats with decremental LTP. These data suggest that the regulated expression of APP isoforms is part of the tanscriptional response associated with the enduring forms of synaptic plasticity and is altered with age. Whereas the level of alpha-synuclein mRNA is not apparently modified in normal LTP, it may reflect a mechanism of apoptotic cell death in aging that is in part responsible for decremental synaptic plasticity. PMID- 11918290 TI - Genetic approach to variability of memory systems: analysis of place vs. response learning and fos-related expression in hippocampal and striatal areas of C57BL/6 and DBA/2 mice. AB - C57 and DBA mice were trained in a crossed maze to assess possible strain differences in place or response learning as a function of training duration (8 or 17 days) and extramaze cueing conditions. The first condition consisted of a diffuse visually cued environment (rich cueing). The second was the same plus an explicit visual cue marking the direction of the baited arm (rich cueing plus cue). The third was a featureless environment (poor cueing). During training, mice were released from the south arm and rewarded in the east arm. Probe trials on which mice were released from the north arm and allowed to choose either the east (place learning) or the west (response learning) arm were given either on the ninth (PT1) or the eighteenth (PT2) days. Strain x context differences in the activation of the dorsal hippocampus and the dorsolateral striatum were examined by analyzing Fos expression following each probe trial. Results first showed that C57 were essentially place-learners, whereas no learning modality was predominant in DBA, except on the PT2 run with the explicit cue available. Examination of Fos expression in C57 trained under "rich cueing" and "rich cueing plus cue" conditions revealed a strong and parallel increase of immunoreactivity in the hippocampus and dorsolateral striatum following PT1 that decreased under PT2. In that strain, the similar time-course variation of Fos expression in both areas suggests a simultaneous involvement of hippocampal- and striatal-based learning mechanisms, even if those controlled by the hippocampus were prevailing on those controlled by the dorsolateral striatum. In DBA mice, however, the absence of any preferential learning modality was associated with 1) a consistent hippocampal activation persistent across probe trials, and 2) a global superior activation of the dorsolateral striatum. Distinct patterns of Fos expression were therefore associated with every strain-specific learning modality. In each strain, however, each modality was found to be remarkably stable, whatever the training duration and the cueing conditions. PMID- 11918292 TI - Insights into immediate-early gene function in hippocampal memory consolidation using antisense oligonucleotide and fluorescent imaging approaches. AB - In the 14 years since it was discovered that specific genes could be dynamically regulated in the brain by neural activity, there has been a substantial research focus attempting to understand the role immediate-early genes (IEGs) play in various brain functions. This article examines the involvement of IEGs in hippocampal synaptic plasticity and in memory consolidation processes performed by the hippocampus. Studies employing conventional IEG detection methodologies and a novel gene-imaging approach that provides temporal and cellular resolution (cellular compartment analysis of emporal activity by fluorescence in situ hybridization or catFISH) provide evidence supporting the assertion that IEG expression reflects the integration of information processed by hippocampal neurons. However, IEG expression is not merely correlated with neural activity, but also plays a pivotal role in stabilizing recent changes in synaptic efficacy. As such, localized disruption of IEGs Arc or c-fos by intrahippocampal administration of antisense oligonucleotides or germline disruption of the IEGs c fos, tissue plasminogen activator, or zif268 impairs consolidation of long-term memory formation, without affecting learning or short-term memory. Further investigation into the expression and function of IEGs using catFISH and antisense approaches will likely increase understanding of the molecular and cellular bases of information processing involving the hippocampus. PMID- 11918291 TI - Aniracetam improves contextual fear conditioning and increases hippocampal gamma PKC activation in DBA/2J mice. AB - DBA/2J (D2) mice display poor contextual learning and have less membrane-bound hippocampal protein kinase C (PKC) compared with C57BL/6 (B6) mice. Aniracetam and oxiracetam were previously shown to improve contextual learning in D2 mice and increase PKC activity. This study investigated a possible mechanism for learning enhancement by examining the effects of aniracetam on contextual fear conditioning and activation of the y isoform of PKC (gamma-PKC) in male D2 mice. In comparison to animals treated with vehicle only (10% 2-hydroxypropyl-beta cyclodextrin), mice treated with aniracetam (100 mg/kg) 30 min prior to fear conditioning training demonstrated significantly improved contextual learning when tested 30 min and 24 h after training. This corresponded with a significant increase in activated, membrane-bound hippocampal gamma-PKC 30 min after training. No increase in learning or gamma-PKC was found 5 min after training. These results suggest an altered time course of activation of gamma-PKC in response to treatment with aniracetam, which improves learning in D2 mice. PMID- 11918293 TI - Efficacy of ex vivo OPG gene therapy in preventing wear debris induced osteolysis. AB - Aseptic loosening of prosthetic implants remains a serious orthopaedic problem and the greatest limitation to total joint arthroplasty. Central to the etiology of aseptic loosening is periprosthetic osteolysis at the bone-implant interface, which is caused by wear debris-induced inflammation. This inflammation produces the critical osteoclast differentiation factor RANKL, which directly stimulates osteoclastogenesis and osteoclastic bone resorption. A dominant factor known to counteract this process is the natural RANKL receptor antagonist protein OPG. Here we explore the potential of ex vivo OPG gene therapy for aseptic loosening by evaluating the eflicacy of stably transfected fibroblast-like synoviocytes (FLS) expressing OPG in preventing wear debris-induced osteoclastogenesis, in a mouse calvaria model. Although the stably transfected fibroblasts produced small amounts of OPG (0.3 ng/ml/72 h/10(6) cells), this protein was very effective in preventing osteoclastic resorption as determined in a bone wafer assay. More importantly. implantation of 10(7) FLS-OPG, together with 30 mg of Ti wear debris, onto the calvaria of mice, completely inhibited osteoclastogenesis 3 days after surgery. Animals given FLS-LacZ control cells, which persisted for 3 days as determined by X-gal staining, together with the Ti particles, had a 6-fold increase in osteoclastogenesis compared to controls without Ti. This increased osteoclastogenesis was completely inhibited by the FLS-OPG, as osteoclast numbers in the calvaria of these animals were similar to that seen in the SHAM controls. PMID- 11918295 TI - Outcomes of surgical management of total HIP replacement in patients aged 65 years and older: cemented versus cementless femoral components and lateral or anterolateral versus posterior anatomical approach. AB - This observational study compared the outcomes of 271 cases of hip osteoarthritis receiving primary total hip replacement (patients 65 years of age and older) from numerous surgeons in 12 Baltimore region hospitals from 1991-1993. The independent variables studied were: (a) totally non-cemented prostheses (non cemented femoral component, non-cemented acetabular component) versus hybrid prostheses (cemented femoral component, non-cemented acetabular component), and (b) lateral or anterolateral surgical approach versus posterior surgical approach. Outcomes included complications during the initial hospitalization, hospital length of stay, hospital cost, readmission, and reported and/or observed physical, instrumental, neuromuscular and affective functioning and pain at 2, 6, and 12 months post surgery. Results indicated that, while the totally non cemented prosthesis was more costly, there were no statistically significant differences in clinical or functional outcomes between the non-cemented and the hybrid prostheses up to 12 months post surgery. Also, while the posterior surgical approach was associated with a non-statistically significant higher rate of dislocation, overall, there was improved function and reduced pain in the first 12 months post-surgery associated with this approach. PMID- 11918294 TI - Tumor necrosis factor-alpha mediates polymethylmethacrylate particle-induced NF kappaB activation in osteoclast precursor cells. AB - Tumor necrosis factor-alpha (TNF) is a potent osteoclastogenic cytokine that has a fundamental role in the pathogenesis of implant particle-induced osteolysis. The nuclear transcription factor NF-kappaB mediates TNF signaling and this transcription complex is necessary for osteoclastogenesis. Because polymethylmethacrylate (PMMA) particles cause osteolysis, we reasoned the PMMA would induce NF-kappaB activation. In fact, we find that exposure of osteoclast precursors, in the form of colony stimulating factor-1 (CSF-1) dependent murine bone marrow macrophages, to PMMA particles prompts nuclear translocation and activation of NF-kappaB. Supershift assays confirm the presence of the p50 and p65 NF-kappaB subunits in the activated transcription factor. Particle-induced NF kappaB activation is equal in both wild type and LPS- hyporesponsive cells indicating that the phenomenon does not represent endotoxin contamination. A soluble, competitive inhibitor of TNF (huTNFr:Fc) dampens particle-directed NF kappaB activation and this response is also abrogated in TNF-/- osteoclast precursors. Thus, PMMA particle activation of NF-kappaB is a secondary event resulting from enhanced TNF expression and is independent of LPS contamination. PMID- 11918296 TI - Geometric analysis of selected press fit prosthetic systems for proximal humeral replacement. AB - The ability of a prosthetic system to replicate a wide range of normal anatomy appears dependent in part to its capacity to produce variable prosthetic geometries. Several modern designs have recently been developed in order to provide multiple prosthetic options. The purpose of this study was to compare the geometry of select press fit prosthetic systems in terms of their ability to match normal three-dimensional geometry of the proximal humerus. The anatomy of 60 humeral specimens from 30 cadavers known from CT data and computer-aided design (CAD) analysis was compared to a 1996 database of four conventional shoulder prosthetic systems (fixed inclination angle, one medial-lateral offset position per head) and one so-called anatomic system (variable inclination angles, multiple head offset positions). The prosthetic system with greatest geometric options allowed for a significantly better replication of the anatomy. Average displacement of the center of rotation was 2.1 mm compared to 9.7 mm for the other systems combined. Reduction in surface arc was 12 degrees compared to 32 degrees. The most problematic feature of conventional prosthetic systems in terms of replicating normal humeral anatomy is the gap created by the prosthetic collar and Morse taper. PMID- 11918297 TI - Low molecular weight heparin aggravates infectious arthritis triggered by Staphylococcus aureus. AB - PURPOSE: Staphylococcus aureus is responsible for the majority of wound infections in clean surgical procedures that involve implantation of foreign material, grafts or prosthetic devices. The aim of the study was to assess the effect of low molecular weight heparin on the development and progression of S. aureus arthritis. MATERIALS AND METHODS: The murine model of hematogenously acquired septic arthritis was used injecting intravenously toxic shock syndrome toxin-1 (TSST-1) producing S. aureus of LS-1 strain. Mice lacking prosthetic implants were treated with intraperitoneal injections of low molecular weight heparin, used routinely as anti-thrombotic prophylaxis following joint prosthetic surgery. Evaluation of arthritis was performed clinically and histopathologically. In addition, the effect of low molecular weight heparin on T cell dependent and independent inflammation was assessed. RESULTS: Seven days after inoculation with bacteria 18 out of 19 low molecular weight heparin treated mice displayed clinical symptoms of arthritis as compared to 9 out of 23 control animals (p < 0.05), and the severity of arthritis, expressed as arthritic index, was 2.6+/-0.5 versus 1.6+/-0.5 (p = 0.05). The histopathological examination confirmed the clinical findings showing that both inflammation and joint destruction were more substantial in heparin treated animals. CONCLUSION: Our findings indicate that the routine anti-coagulation treatment with heparin contributes to more severe course of joint infection. PMID- 11918298 TI - Changes in human knee ligament stiffness secondary to osteoarthritis. AB - Stiffness of the medial (MCL) and lateral (LCL) collateral ligaments was compared between a group of 10 patients undergoing total knee arthroplasty for varus degenerative osteoarthritis (OAP), a group of 10 osteoarthritic cadaveric knees (OAC), and a group of 10 non-osteoarthritic cadaveric knees (NOA). A load elongation curve was obtained for the medial and lateral compartments of each knee using an instrumented Moreland spreader. A strain gage (SG) was attached to the spreader handle and strain was calibrated to load applied against the spread distance. In extension, medial compartment stiffness of the OAP, OAC, and NOA groups was 60.7+/-16, 52.8+/-9.3 and 21.4+/-5.0 N/mm, respectively. Lateral compartment stiffness in extension was 29.2+/-9.2, 33.3+/-10.3 and 19.5+/-5.3 N/mm, for OAP, OAC, and NOA, respectively. Differences in stiffness between the OAP and OAC groups were not statistically significant (p > 0.05). However, the osteoarthritic groups (OAP and OAC) demonstrated a statistically significantly (p < 0.05) increase in ligament stiffness when compared to the NOA group. Following knee arthroplasty, stiffer medial structures in extension may lead to flexion contracture and accelerated polyethylene wear. Adequate bone resection, in conjunction with soft tissue release may alleviate the threefold increase in stiffness observed in the medial compartment secondary to OA without jeopardizing joint stability. PMID- 11918300 TI - Wrist kinetics after scapholunate dissociation: the effect of scapholunate interosseous ligament injury and persistent scapholunate gaps. AB - The purpose of this study was to investigate the effects of cutting of the scapholunate interosseous ligament (SLIL) and persistent widening of the scapholunate (SL) joint on changes in moment arms of the principal wrist motor tendons. In seven fresh frozen cadaveric upper extremities, excursions of the extensor carpi radialis longus (ECRL) and brevis (ECRB), extensor carpi ulnaris (ECU), flexor carpi radialis (FCR), and flexor carpi ulnaris (FCU) were recorded simultaneously with wrist joint angulation during wrist flexion-extension and radioulnar deviation. Tendon excursions were measured in intact wrists, then in the wrists with complete SLIL sectioning and in those with moderate or severe persistent SL joint widening. The data were converted to moment arms of the tendons. The results showed that moment arms of the ECRL and ECRB tendons after SLIL sectioning were, respectively, 110+/-6% and 105+/-3% of those in the intact wrist. In the wrists with moderate or severe SL joint widening, moment arms of the flexors significantly increased (P < 0.01 and P < 0.001, respectively). During radioulnar deviation, moment arms of the ECRL, ECRB, ECU, and FCU tendons decreased after SLIL sectioning and the SL joint widening. However, moment arms of the FCR tendon significantly increased 122+/-23% after the SLIL section, 133+/ 28% after the moderate SL joint widening, and 138+/-24% after the severe SL joint widening compared with those of the intact wrists. This study demonstrated that integrity of the SLIL and appropriate SL joint space are important for mechanics of wrist motor tendons. Loss of integrity of the SLIL and persistent SL joint widening increase mechanical effects of the radial side wrist motor tendons, which may contribute to the pathomechanics of scaphoid malrotation, scapholunate advanced collapse, and early osteoarthritis in the radioscaphoid joint interface seen in SL dissociation. The results also suggest that reduction of the displaced SL joint is imperative to the recovery of wrist kinetics after SL dissociation. PMID- 11918299 TI - Abnormal knee joint position sense in individuals with patellofemoral pain syndrome. AB - The purpose of this cross-sectional study was to compare knee joint position sense (JPS) in 20 individuals with and 20 without patellofemoral pain syndrome (PFPS). Five active tests with ipsilateral limb matching responses were performed at 20 degrees and 60 degrees flexion under non-weightbearing conditions, and at 40 degrees flexion under uni- and bi-lateral weightbearing conditions. The response errors were calculated as the difference between each target and response position (accuracy) and the standard deviation of these differences (reliability). JPS was: (i) significantly less accurate and less consistent in the knees with PFPS during both the non-weightbearing and weightbearing tests when compared to the control subject knees; (ii) less accurate when the symptomatic and asymptomatic knees of the 12 uni-lateral PFPS subjects were compared and (iii) less accurate in the asymptomatic knees of the uni-lateral PFPS subjects and knees of the control subjects. The maximum intensity of pain experienced during each knee JPS test was not correlated to any of the JPS test results. The results confirm abnormal knee joint proprioception in individuals with PFPS. Although it cannot be determined whether the abnormality precedes or follows the development of PFPS, the results support including proprioceptive reeducation in management of PFPS. PMID- 11918301 TI - Plantar impact causing midfoot fractures result in higher forces in Chopart's joint than in the ankle joint. AB - PURPOSE: Force effect (impact, extent of foot compartment deformation) and result (fracture pattern) for midfoot fractures in car occupants is known. An analysis of the processes in the foot was intended to improve car safety. MATERIALS AND METHODS: Eleven fresh, unfrozen, unpreserved intact human cadavers (age: 36.8 (16 61) years, gender: male, race: Caucasian) were studied 24-72 h after death. In 3 cadavers (5 feet) the experimental design was established: entire cadaver fixed on a special tray in supine position, pendulum with bar impactor hitting the foot plantar to Lisfranc's joint. A custom-made pressure sensor was inserted in the ankle (A), talonavicular (TN) and calcaneocuboid (CC) joints (resolution: 1 cm2, sampling rate: 500/s). RESULTS: Sixteen feet were measured: midfoot fractures were induced in 11 feet. The maximum pressure amounted to 1.22-2.55 MPa (2.04+/ 0.412) at 0.005 0.195 s (0.067+/-0.059) after impact. The maximum pressure occurred in 8 (50%) cases in the ankle, in 7 (44%) of the TN and 1 (6%) of the CC joints. A comparison of the first 200 pressure samples after impact of all sensor fields resulted in higher forces in Chopart's joint than in the ankle (t-test: p < 0.001). These force differences were higher in cases with midfoot fractures (mixed model analysis of variance: p = 0.003). CONCLUSION: Due to considerable forces in Chopart's joint we recommend a modification of the actual crash test dummy lower extremity model with an additional load cell that detects forces in the longitudinal direction of the foot axis. PMID- 11918302 TI - Upregulation of basal TGFbeta1 levels by EMF coincident with chondrogenesis- implications for skeletal repair and tissue engineering. AB - Members of the TGFbeta/BMP gene family regulate cartilage and bone development. These genes are re-expressed in bone repair and are thought to mediate chondro- and osteoprogenitor cell differentiation. These observations have led to a therapeutic strategy of introducing these growth factors into experimental cartilage and bone defects. Therapeutic efficacy, however, has been limited by diffusion or inactivation of these growth factors from the desired site and by the inability to deliver sustained concentrations of growth factors. This study demonstrates an increase in basal TGFbeta mRNA and protein levels in association with chondrogenic differentiation in endochondral ossification. mRNA is increased by 158%; protein by 23%, and cells immunopositive for TGFbeta by 343% at maximal TGFbeta expression. Importantly, the pattern of TGFbeta expression is preserved throughout the developmental sequence. Our data suggest that the exposure to a specific electromagnetic field (EMF) enhances, but does not disorganize, chondrogenesis and endochondral calcification as well as the normal physiologic expression of TGFbeta. The ability to increase TGFbeta at a moderately low dose for sustained periods of time without disorganizing its physiology suggests the ability to establish temporal concentration gradients of growth factors for the purpose of stimulating skeletal repair. PMID- 11918303 TI - Photodynamic treatment has chondroprotective effects on articular cartilage. AB - Osteoarthritis (OA) is a disabling joint disease for which there is currently no cure. It is characterized by the destruction of articular cartilage. One strategy that is being explored for protecting the cartilage in OA is the administration of transforming growth factor-beta, which in vitro antagonizes cartilage degradation initiated by catabolic stimulants such as interleukin-1 (IL-1). The problems associated with selective delivery of the growth factor to chondrocytes, undesirable side-effects on joint tissues, and short biological half-life have led us to explore modalities aimed at activating transforming growth factor-beta that is stored in the cartilage as latent complexes. Photodynamic therapy is a two-step protocol of tissue sensitization with a light-activatable chemical called a photosensitizer followed at some interval by irradiation with the appropriate wavelength visible light. Biological effects are typically elicited through oxygen-dependent photochemistry without heat generation. Transforming growth factor-beta1 can be activated by oxidative mechanism(s), prompting us to explore whether photodynamic technology can be harnessed to modulate cartilage metabolism. Disks of bovine articular cartilage were photosensitized by incubation with a chlorin(e6)-succinylated polylysine conjugate and irradiated with 1-2 J/cm2 red light (lambdamax = 671 nm). This two-step regimen dramatically inhibited IL-1-stimulated proteoglycan degradation and concomitantly increased latent and active transforming growth factor-beta1 in culture medium. This research may lead to the development of minimally invasive photodynamic therapy in which light is delivered to locally activate a chondroprotective program in photosensitized cartilage in the context of OA. PMID- 11918304 TI - Quantitative cartilage imaging of the human hind foot: precision and inter subject variability. AB - Alterations of ankle cartilage are observed in degenerative and inflammatory joint disease, but cartilage cannot be directly visualized by radiography. The purpose of this study was therefore to analyze the feasibility and precision of quantitative cartilage imaging in the human hind foot (talocrural, talotarsal, and intertarsal joints), and to report the inter-subject variability for cartilage volume, thickness and surface areas. The feet of 16 healthy volunteers were imaged using a 3D gradient-echo magnetic resonance imaging sequence with water-excitation. After interpolation to a resolution of 1 x 0.125 x 0.125 mm3 the cartilage plates were segmented, and the cartilage volume, thickness, and surface areas determined. The precision (four repeated measurements) was examined in eight volunteers, the RMS average CV% being 2.1% to 10.9% in single joint surfaces, and < or = 3% for the cumulative values of all joints. The mean cartilage thickness ranged from 0.57+/-0.08 (navicular surface) to 0.89+/-0.19 mm (trochlear surface for tibia). In conclusion this study shows that it is feasible to quantify thin cartilage layers in the hind foot under in vivo imaging conditions, and that the precision errors are substantially smaller than the inter-subject variability in healthy subjects. PMID- 11918305 TI - The characterization of versican and its message in human articular cartilage and intervertebral disc. AB - Splicing variation of the versican message and size heterogeneity of the versican core protein were analyzed in human articular cartilage and intervertebral disc. Splicing variation of the message was studied by PCR analysis to detect the presence or absence of exons 7 and 8, which encode large chondroitin sulfate attachment regions. At all ages in normal cartilage from the third trimester fetus to the mature adult, the presence of the versican isoform possessing exon 8 but not exon 7 (V1) could be readily detected. The message isoforms possessing neither exon 7 nor 8 (V3) or both exons 7 and 8 (V0) were only detectable in the fetus, and the isoform possessing only exon 7 (V2) was never detected. In osteoarthritic cartilage and in adult intervertebral disc the versican message pattern was the same as that observed in the normal adult with only the isoform possessing exon 8 being detected. Core protein heterogeneity was studied by immunoblotting following enzymic removal of the glycosaminoglycan chains from the proteoglycan, using an antibody recognizing the globular G1 region of versican. All articular cartilage extracts from the fetus to the mature adult contained multiple core protein sizes of greater than 200 kDa. The adult cartilage extracts tended to have an increased proportion of the smaller sized core proteins and osteoarthritic cartilage possessed similar core protein sizes to the normal adult. In contrast, intervertebral disc at all post-natal ages showed a greater range of size heterogeneity with a prominent component of about 50 kDa. The abundance of this component increased if the samples were treated with keratanase prior to analysis, suggesting that the G1 region of versican in disc can be substituted with keratan sulfate. The increased presence of versican in the disc relative to articular cartilage may suggest a more pronounced functional role for this proteoglycan, particularly in the nucleus pulposus. PMID- 11918306 TI - Hydroxyapatite crystals as a local delivery system for cisplatin: adsorption and release of cisplatin in vitro. AB - This study describes the characteristics of the in vitro binding and release of the anti-tumor drug cisplatin by slurries of synthetic hydroxyapatite crystals carried out in aqueous media. The adsorption of cisplatin by slurries of hydroxyapatite and its release were found to depend significantly on the ionic composition of the aqueous media used. At a constant pH of 7.4, significantly more cisplatin is adsorbed by the hydroxyapatite crystals in the slurry from a chloride-free phosphate buffered solution or a Tris buffered solution than from a buffered phosphate solution containing chloride ions. The amount of hydroxyapatite-bound cisplatin desorbed into solution was also progressively increased as a function of the increasing concentration of chloride in the equilibrating solution. Very little hydroxyapatite-bound cisplatin was released from the crystals in either a Tris or phosphate buffer. These results suggest that it is the hydrated derivatives of cisplatin which are involved in the adsorption of cisplatin by hydroxyapatite crystals. The adsorption data can be expressed as a Freundlich isotherm from which the association constant can be calculated. The rate of release of cisplatin bound to crystals of hydroxyapatite is relatively slow even at the maximum concentration of chloride ions in the phosphate buffer. Approximately 33% of the total cisplatin bound to the crystals of hydroxyapatite was released after 4.25 days. An additional 15% of the remaining cisplatin bound to the hydroxyapatite cyrstals was released after an additional equilibration with fresh buffer for two weeks (58% of the total cisplatin originally bound). These findings suggest that cisplatin bound to slurries of hydroxyapatite crystals may be useful in the local treatment of malignant tumors. PMID- 11918307 TI - Estrogen regulation of growth and alkaline phosphatase expression by cultured human bone marrow stromal cells. AB - Estrogen has been reported to regulate the growth and differentiation of cultured murine osteoprogenitor cells in bone marrow stroma. This study tested the ability of 17beta-estradiol (E2) to regulate growth and expression of alkaline phosphatase (ALP), an osteoblastic differentiation marker, in strains of normal human bone marrow stromal cells derived from different donors. In eight strains examined, E2 at 1 and 10 nM produced at most modest effectxs on growth and ALP activity. Growth inhibition, seen in 4 of the 8 strains, was more common than stimulation (2 of the 8 strains); the greatest observed E2 effect was an inhibition of ca. 50%. E2 altered ALP activity less dramatically than cell growth. Differences from control in total ALP per culture were seen in only two strains: one was a reduction, one an increase. Colony forming assays were used to determine if E2 changed the proportion of ALP-expressing cells in marrow stromal cell cultures. In contrast to growth experiments, ALP expression under colony forming conditions (200 cells per 35 mm-diameter well) was dependent on the type of serum supplementation used. Under permissive conditions using medium supplemented with 10% charcoal-treated fetal bovine serum, 10 nM E2 increased the number of ALP-positive colonies (cfu-ap) but not the total number of colonies formed (cfu-f). When cells cultured in the presence or absence of 10 nM E2 were replated at colony forming densities, significantly higher proportions of cfu-ap were found in 2 of 6 strains examined, while pretreatment with E2 affected the number of cfu-f in only 1 of the 6 strains. Similar results were obtained when colony formation was carried out in the presence of dexamethasone and ascorbate, although these agents themselves increased the formation of both cfu-f and cfu ap. These results show that the direct effects of E2 on human marrow stromal cells are small and vary depending on the cell strain and on the experimental conditions; however, the E2 actions observed in this study were consistent with reports that E2 exerts direct actions on osteoblasts and osteoblast progenitor cells that favor rather than suppress their phenotypic expression. PMID- 11918308 TI - Identification of the metalloproteinase stromelysin in the physis. AB - For long bone growth to occur, calcification of the matrix must commence in the lower hypertrophic zone of the growth plate. It is generally accepted that physeal proteoglycans help regulate mineralization, and that at least in vitro, intact proteoglycans can inhibit mineralization. Thus degradation of proteoglycan may be a necessary step prior to calcification. Previous work in our laboratory has demonstrated the presence of neutral metallo-proteases in the growth plate with highest levels in the hypertrophic zone, where calcification occurs. Stromelysin (MMP-3) is a connective tissue matrix-degrading enzyme. It was formerly known as proteoglycanase and is generally considered to be one of the major proteoglycan degrading enzymes in cartilage. Stromelysin is implicated in cartilage destruction in osteoarthritis and may also be involved in tissue remodeling in the physis. Our goal was to determine if the neutral protease previously reported by the authors in the physis was stromelysin. In this study we used Western blots and antibodies to stromelysin and to the stromelysin cleavage site in aggrecan, the most common form of proteoglycan, to demonstrate the presence of stromelysin in the bovine physis. When an antibody raised against the stromelysin cleavage site of aggrecan (FVDIPEN) was incubated with a Western blot, which had been run with aggrecan extracted from bovine physes, a positive reaction resulted. This suggests that there is stromelysin degradation in vivo in the physis. Two different polyclonal antibodies to stromelysin gave positive results on Western blots of purified media from growth plate cultures indicating that stromelysin is produced in vitro in culture. These antibodies also reacted with active stromelysin. The presence of stromelysin in the physis implicates it in physeal physiology. The concentration of its activity in the lower hypertrophic zone and zone of provisional calcification suggests that it may be particularly important in mineralization. PMID- 11918309 TI - Nitric oxide induces osteoblast apoptosis through the de novo synthesis of Bax protein. AB - Nitric oxide (NO) plays a crucial role in the physiological and pathophysiological regulations of osteoblast functions. This study is designed to evaluate the toxic effects of NO released by sodium nitroprusside (SNP), an NO donor, on neonatal Wistar rat calvarial osteoblasts from the analyses of cell viability, alkaline phosphatase (ALP) activity, cell morphology, apoptotic cells, terminal deoxynucleotidyl transferase-mediated dUTP nick end-label (TUNEL) assay, DNA ladder, and immunocytochemistry and Western blot for proapoptotic Bax protein. SNP increased the levels of nitrite, an oxidative product of NO, in the culture medium of osteoblasts in concentration- and time-dependent manners, and altered cell morphologies to round and shrinkage shapes. Administration of osteoblasts with SNP resulted in concentration- and time-dependent decreases of cell viability and ALP activity. Analysis of apoptotic cells revealed that SNP increased the percentages of osteoblasts processing apoptosis. Analyses of TUNEL and DNA ladder showed that SNP caused DNA fragmentation. Pretreatment with cycloheximide, an inhibitor of protein synthesis, partially blocked SNP-induced osteoblast apoptosis. Imunocytochemical and immunoblotting analyses revealed that SNP increased Bax protein in osteoblasts. This study suggests that SNP could increase the levels of NO in osteoblasts, and cause osteoblast apoptosis possibly through the de novo synthesis of proapoptotic Bax protein. PMID- 11918311 TI - Alpha-smooth muscle actin is expressed by fibroblastic cells of the ovine anterior cruciate ligament and its free tendon graft during remodeling. AB - Contractile fibroblastic cells expressing the alpha-smooth muscle actin isoform, so-called myofibroblasts, have been identified to play a possible role during the healing of the medial collateral ligament by means of restoring the tissues in situ strain via extracellular matrix contraction. Recently, these cells have also been identified to be a normal part of the human anterior cruciate ligament. It has been hypothesized that myofibroblasts play a role in the wrinkling of the extracellular matrix. The goal of the present study was to identify myofibroblasts in the intact ovine anterior cruciate ligament and a free autologous tendon graft during remodeling after anterior cruciate ligament reconstruction. In 36 mature merino sheep the anterior cruciate ligament was replaced with an ipsilateral Achilles tendon split graft. Midsubstance tissue samples were immunostained for alpha-smooth muscle actin at 6, 9, 12, 24, 52, and 104 weeks. Myofibroblasts were identified in the intact ovine anterior cruciate ligament as well as in the Achilles tendon graft prior to implantation. During remodeling the first myofibroblasts were found at six weeks within newly formed fiber bundles. At 24, 52, and 104 weeks myofibroblast distribution and cell density were similar to those of the intact ovine anterior cruciate ligament. These findings indicate that alpha-smooth muscle actin containing fibroblastic cells are a regular part of the intact as well as the remodeled anterior cruciate ligament. There is evidence that myofibroblasts may be involved in maintaining tissue homeostasis in the mature ligament e.g., by means of crimp formation. The presence of these cells during the early remodeling may further indicate that alpha-smooth muscle actin containing fibroblastic cells are involved in the earliest stages of fiber bundle formation. The role and function of this special cell type for the anterior cruciate ligament needs to be further clarified. PMID- 11918310 TI - A new animal model of femoral head osteonecrosis: one that progresses to human like mechanical failure. AB - Existing animal models of femoral head osteonecrosis, while displaying varying levels of concordance with early histopathologic features of the human disorder, generally fail to progress to end-stage mechanical collapse. A new animal model of osteonecrosis is here introduced, utilizing the emu (Dromaius novaehollandie). These animals' bipedality and their high activity level represent a much more challenging biomechanical environment to the hip than seen in quadrupedal models of this disorder. Femoral head osteonecrosis was induced surgically, using a combination of ischemic (vessel ligation) and cryogenic (liquid nitrogen) insults. Of nineteen emus allowed free-roaming pen activity to study the natural history of such lesions, eighteen progressed to an osseous structural failure, sixteen of them developing incapacitating lameness at an average time point 11.75 weeks after the surgical insult. Histologically, the animals showed close concordance with both the early- and late-stage human pathology, in six cases even to the point of developing a crescent sign. Because of its large physical size and its consistent progression to mechanical collapse, the emu appears to offer a unique opportunity for the near-human-scale study of surgical interventions to forestall femoral head collapse. Toward this end, various directions for model refinement are outlined. PMID- 11918313 TI - Chronic anterior cruciate ligament deficiency is associated with increased anterior translation of the tibia during the transition from non-weightbearing to weightbearing. AB - Translation of the tibia relative to the femur was measured while a group of subjects with normal knees and group with anterior cruciate ligament (ACL) tears underwent transition from non-weightbearing to weightbearing stance. Subjects were positioned in the Vermont knee laxity device (VKLD) with muscles relaxed and the limb segment and compressive joint load offset (non-weightbearing). A lateral radiograph of the knee, with the posterior aspects of the femoral condyles superimposed, was obtained to document the position of the tibia relative to the femur. Immediately after, a compressive load equal to 40% of bodyweight was applied to each foot, and a second radiograph was obtained to document the change in position of the tibia relative to the femur. The transition from non weightbearing to weightbearing produced a significant increase of anterior translation of the tibia relative to the femur (mean; 3.4 mm) for the subjects with ACL tears compared with the contralateral normal knees (0.8 mm). Similarly, there was a significant increase in anterior translation of the tibia for the subjects with ACL tears compared to the group of subjects with normal knees (1.2 mm). The fourfold increase in anterior translation of the tibia for the knees with ACL tears compared to the contralateral side is a concern because it is substantially greater than the 95% confidence limits of the side-to-side differences in anterior-posterior knee laxity measured from subjects with normal knees. This observation could explain, at least in part, one of the mechanisms that initiates damage to the meniscus and articular cartilage in subjects that have suffered an ACL tear. PMID- 11918312 TI - Collagen expression and biomechanical response to human recombinant transforming growth factor beta (rhTGF-beta2) in the healing rabbit MCL. AB - We investigated biomechanical and collagen expression in a healing bilateral rabbit medial collateral ligament (MCL) model to human recombinant transforming growth factor beta (rhTGF-beta2) at three and six weeks. Each rabbit had rhTGF beta2 in a bioabsorbable pellet administered in one side, with the contralateral side serving as control (no rhTGF-beta2). All MCL healed with rhTGF-beta2 producing a profoundly increased scar mass at three weeks which decreased in size toward control at six weeks. In-situ hybridization demonstrated collagen expression (type I and III) no different than control at three weeks, but by six weeks elevated expression of type I was seen. Biomechanical analysis at three weeks showed no effect of rhTGF-beta2 on structural properties. However, at six weeks rhTGF-beta2 significantly inhibited both the maximum load (p < 0.05) and energy absorbed (p < 0.05) with no change in stiffness. Despite increased type I collagen expression and profound increase in early scar mass, rhTGF-beta2 did not improve the structural properties. Whether the dose or mode of delivery is responsible for decline in structural properties cannot be determined in this design. We hypothesize investigations of healing ligaments to cytokines should have biologic and biomechanical properties correlated in the same study at a minimum of two time points. PMID- 11918314 TI - The effects of graft width and graft laxity on the outcome of caprine anterior cruciate ligament reconstruction. AB - We studied how initial graft size and initial graft laxity affected the biomechanics of anterior cruciate ligament (ACL) reconstruction at six months. Sixteen goats had bilateral reconstructions staged eight weeks apart. Autografts 4 and 7 mm wide were taken from the central patellar tendon (PT). Lax grafts were created by adding 4 mm slack to the graft before fixing. We reconstructed each joint using a combination of width and laxity treatments. Both factors were changed for the contralateral joint and all combinations appeared with equal frequency. At six months we measured the joint extension limit, anterior posterior (AP) translation, and osteoarthritic changes. The grafts were then tested to failure to determine their mechanical properties. After six months the difference in initial treatments had disappeared: there was no difference in graft cross-section due to the different initial widths and there was no difference in joint AP translation due to the initial graft laxity. We did observe that wide grafts were associated with a block to extension, decreased joint AP translation, and increased articular cartilage damage and osteophyte formation. While AP translation was reduced, it was correlated with decreased extension, possibly indicating an increase in scar tissue formation rather than a more functional graft. Neither graft width nor graft laxity produced differences in any graft mechanical properties. This suggests that the use of larger grafts to prevent increased AP translation has undesirable complications. Ultimately, we conclude that neither of these surgical treatments strongly affects the biomechanical result of caprine ACL reconstruction. PMID- 11918315 TI - Medial collateral ligament autografts have increased creep response for at least two years and early immobilization makes this worse. AB - Recent evidence has shown that 10-40% of knee joints reconstructed with soft tissue autografts have a recurrence of abnormal joint laxity over time. One possible explanation is the "stretching out" (or unrecovered creep) of the graft tissue. To test in vitro creep and creep recovery of fresh anatomic ligament autografts in an extra-articular environment, 16 rabbits underwent an orthotopic medial collateral ligament (MCL) autograft procedure to one hindlimb. Three subgroups of animals had either unrestricted cage activity for 1 year (n = 5) or 2 years (n = 5) or pin-immobilization for the first 6 weeks followed by cage activity for the remainder of 1 year (n = 6). Following laxity measurements, to test their creep response, isolated MCL grafts were cyclically and then statically creep tested in vitro at 4.1 MPa, allowed to recover at zero load for 20 min, and finally elongated to failure. Due to differences in cross-sectional area between the grafts and normal MCLs, two normal control groups were tested: stress-matched tested at 4.1 MPa (16.2 N; n = 7) and force-matched tested at 29.1 N (7.1 MPa; n = 6). Ligament grafts had normal laxity but significantly increased creep and decreased creep recovery compared to normal MCLs after I and 2 years of healing (p < 0.0004). Graft failure stress was also significantly less than normal (p < 0.0001). Immobilized grafts had significantly greater creep compared to non-immobilized grafts at 1 year of healing (p < 0.05). These results support previous observations concerning material inferiority of fresh anatomic rabbit MCL autografts, but add the concept that such grafts also have increased potential to creep with either slower or incomplete recovery when subjected to low stresses in vitro. Joint and ligament laxities in situ were normal in this model, however, suggesting either that in vivo MCL graft stresses are lower than those used here in vitro or that these tissues have other mechanisms by which they can recover their functional length in vivo. PMID- 11918316 TI - Effects of different exposures of hyperbaric oxygen on ligament healing in rats. AB - Hyperbaric oxygen (HBO) is a method of augmenting, intermittently, oxygen availability to tissues. We examined the effect of three different HBO exposures on the healing of experimentally induced ligament lacerations in the right hind limb of 44 male Wistar rats. Animals were divided into four groups after ligament injury: (a) control group, animals breathed room air at 1 ATA (atmosphere absolute) in a hyperbaric chamber for 60 min; (b) HBO treatment at 1.5 ATA for 30 min once a day, (c) HBO treatment at 2 ATA for 30 min once a day, (d) 2 ATA for 60 min once a day. At 14 days post-ligament injury, we compared the ligaments of the four treatment groups for gross appearance, histology and expression of pro alpha(I) mRNA by northern hybridization. Our results indicate that HBO was effective in promoting ligament healing compared to control (p < 0.01). Of these three exposures, HBO at 2 ATA for 60 min was the most effective, resulting in enhanced extra-cellular matrix deposition as measured by collagen synthesis. PMID- 11918317 TI - Delayed tendon reattachment does not reverse atrophy and fat accumulation of the supraspinatus--an experimental study in rabbits. AB - Muscle atrophy and fat accumulation occur after rotator cuff tearing. Whether these changes are reversible after a successful repair is still unknown. Imaging allows only a semi-quantitative assessment of muscle atrophy and fat deposition. As only experimental studies permit a quantification of both, a unilateral detachment and wrapping in a polyvinylidine fluoride membrane of the supraspinatus was done in 22 rabbits. After 12 weeks, the polyvinylidine fluoride membrane was removed in 10 rabbits and the tendon inserted into a bony trough. The opposite shoulder served as control. All rabbits were killed at 24 weeks. Accumulation of fat on the muscle surface as well as between the muscle fascicles and occasionally in the sarcoplasm was increased in both experimental groups when compared to controls (non-reattached 12.8%, reattached 23.5% vs controls 5.4%; all p < 0.005). The muscle atrophy was significant (non-reattached 9.2 ml, reattached 9.1 ml vs controls 10.7 ml; both p < 0.005). Neither atrophy nor fat accumulation were reversed by surgical reattachment of the supraspinatus tendon. On the contrary, the fat accumulation was higher in the reattached group (p < 0.005). The trauma of reattachment, the delay between detachment and reattachment or an insufficient duration of recovery may have contributed to both, the lack of reversibility of supraspinatus atrophy and fat accumulation. PMID- 11918319 TI - An in vivo study of peripheral nerves in continuity: biomechanical and physiological responses to elongation. AB - Using an animal model, the in vivo visco-elastic properties of peripheral nerve in continuity were examined. The nerves were stretched by either 1 cm - Group A (8.8% strain) or 2 cm - Group B (16.1% strain). At constant strain, the stress relaxation curves were plotted. Maximal relaxation was observed in the first 20 min. After 1 h, the decrease in tensions in Groups A and B were 36.8% and 41.8%, respectively. Throughout this hour and for 30 min after the release of tension, regular recordings of nerve blood flow using laser doppler flowmetry and peak nerve conduction velocity were taken. Nerve blood flow was reduced by similar amounts (Group A, 70%; Group B, 78%) by application of traction. On release, however, Group A displayed a reactive hyperaemia (blood flow 151% starting value) while Group B failed to recover (50% starting value at 30 min). Peak nerve conduction velocity was not significantly altered in Group A in response to traction, whereas in Group B the result was a gradual, but significant reduction in peak velocity to 66% starting value. The lack of correlation between blood flow and peak nerve conduction velocity in these studies suggests that ischaemia is not solely responsible for the increased latency. PMID- 11918318 TI - Differential expression of neural cell adhesion molecule (NCAM) after tenotomy in rabbit skeletal muscle. AB - Tenotomy is a commonly encountered event in orthopaedic surgery. In 23 rabbit extensor digitorum longus (EDL) muscles, within 24 h after tenotomy, a marked drop in maximum force production occurred. This was not explainable based on architectural changes and histological examination using standard markers for muscle injury, i.e., haematoxylin and eosin morphology, developmental myosin heavy chain (MHC) immunolabeling, and quantitation of muscle fiber type percentage, area and distribution. The expression of neural cell adhesion molecule (NCAM), a glycoprotein expressed during muscle development was measured as a function of time in these muscles. NCAM expression was increased as early as one day after tenotomy with 2.2+/-1.2% of the fibers showing positive expression. This expression level increased significantly to 15.4+/-15.2% after 7 days and then subsided to 13.2+/-10.6% 21 days after tenotomy. Two-way analysis of variance demonstrated a significant effect of time and a significant time x tenotomy method interaction. These results suggest that tenotomy leads to possible changes in muscle-nerve connections and/or excitation-contraction (EC) coupling. PMID- 11918320 TI - Epidural injection of cyclooxygenase-2 inhibitor attenuates pain-related behavior following application of nucleus pulposus to the nerve root in the rat. AB - Cyclooxygenase-2 (COX-2), the inducible isoform of COX, has been identified as the key enzyme to regulate prostaglandin E2 synthesis in inflammatory conditions. Although it has been reported that COX-2 is present in herniated disc samples obtained from patients, little is known concerning the relationships between COX 2 and painful radiculopathy. The purpose of this study was to evaluate whether epidural injection of COX-2 inhibitor abolishes hyperalgesia induced by nucleus pulposus, which is a pain-related behavior in the rat. Rats, in which nucleus pulposus was relocated on the nerve root, exhibited evidence of mechanical hyperalgesia. Epidural injection of COX-2 inhibitor resulted in decrease in mechanical hyperalgesia 1 h, 3 and 7 days after the epidural injection of COX-2 inhibitor (0.1 mg/kg SC-'236 dissolved in the vehicle). There were no significant differences in sensitivity to thermal noxious stimuli after either application of the nucleus pulposus or epidural injections. These results suggest that prostaglandins and thromboxane, which are produced by COX-2 in inflammatory cells, appear to be related to the inflammatory process produced by application of nucleus pulposus to the nerve root. It is possible that COX-2 plays a significant role in painful radiculopathy following herniated nucleus pulposus. PMID- 11918321 TI - Serum cartilage-derived retinoic acid-sensitive protein (CD-RAP) levels in swarm rat chondrosarcoma. AB - Cartilage-derived retinoic acid-sensitive protein (CD-RAP) is a new protein that was isolated from bovine articular chondrocytes and human melanoma cell lines (melanoma inhibitory protein or MIA). In normal tissue its expression is limited to cartilage, and in morbid tissue to melanoma, chondrosarcoma, and breast cancer. Serum levels of CD-RAP/MIA correlate with the progression of malignant melanoma, but there have been no reports on chondrosarcoma. Here, it was first demonstrated by RT-PCR and immunohistological methods that CD-RAP was expressed in tissue from a Swarm rat chondrosarcoma that was used as an experimental model. The course following tumor transplantation and changes in serum CD-RAP after tumor excision were then observed to investigate whether serum CD-RAP could be used as a marker of tumor activity. Consequently, serum CD-RAP in control rats tended to decrease as the animal grew, whereas it rose in proportion to tumor proliferation in rats that had received a tumor graft. Serum CD-RAP levels dropped rapidly following excision of the tumor in a group of tumor-excised rats. In those rats which had a recurrence following excision of the tumor, serum CD RAP rose prior to the appearance of the tumor. Serum CD-RAP thus sensitively reflected tumor onset and proliferation, so that it appeared to be an effective marker of tumor activity for Swarm rat chondrosarcoma. PMID- 11918322 TI - Clonality studies in giant cell tumor of bone. AB - Genetic studies including chromosome analysis, telomere reduction and telomere activity, DNA microsatellites and loss of heterozygosity (LOH) studies have been performed on giant cell tumor (GCT) of bone however whether this primary skeletal neoplasm represents a monoclonal or polyclonal proliferation is unknown. Utilizing a new assay to study the polymorphic human androgen receptor locus (HUMARA), the ratio of maternal inactive X-chromosome to the paternal inactive X (Lyon hypothesis) is determined via a methylation--specific polymerase chain reaction (PCR) technique to detect X-chromosome polymorphisms. Characterization of the genetic tumorigenesis of this unpredictable neoplasm may lend insight into its biological behavior and offer improvements in therapeutic intervention, as new information emerges regarding osteoclastic bone resorption. Seventeen female patients with giant cell tumor of bone had their DNA harvested and their X chromosome inactivation pattern and polymorphisms determined and compared to control. A polyclonal proliferation pattern was identified in all informative samples studied. PMID- 11918324 TI - Trends in alcohol consumption in undergraduate third level students: 1992-1999. AB - BACKGROUND: Alcohol consumption has increased in the Irish population in recent years. It is not known to what extent the student population has been affected by this increase. AIM: To determine levels of alcohol consumption among undergraduates in one Irish university and identify changes in drinking patterns in the years 1992-1999. METHODS: Information on alcohol use was obtained by anonymous self-completed questionnaire in a stratified random cross-faculty sample of undergraduates in 1992 and 1999. The CAGE questionnaire to determine problem drinking was included in both surveys. RESULTS: A statistically significant (p=0.01) drop in weekly alcohol consumption by males was found, although the proportion of male problem drinkers increased. Consumption for females remained the same. CONCLUSIONS: Findings are contrary to recent figures for drinking patterns in young Irish people in general. The fall in alcohol consumption in male students may be linked to improved male insight into the negative effects of alcohol or to the substitution of cheaper available substances. PMID- 11918325 TI - The future of paediatric surgery outside specialist paediatric hospitals. AB - BACKGROUND: In Ireland, general surgeons provide paediatric surgical services to patients remote from specialist paediatric units. AIM: To review general paediatric surgical services in the Mid-Western Health Board (MWHB) region with a view to informing future policy decisions in Ireland. METHODS: From 1995 to 2000, the paediatric surgical workload at the Mid-Western Regional Hospital was reviewed. Operations performed, level of operating surgeon, morbidity and mortality were recorded. RESULTS: There were 3,166 general paediatric surgical patients cared for by three general surgeons and one urologist, all with paediatric surgical training. There was an increase in day cases (55% to 70%) and operations at which the consultant was the main operator (40% to 67%). There was no mortality and the morbidity rate was less than 1%. CONCLUSIONS: General paediatric surgery and urology is well provided for in the MWHB by appropriately trained surgeons. Lack of opportunity for surgeons in training to obtain general paediatric surgical experience will put this service at risk as the current cohort of surgeons retire. Expansion in specialist paediatric surgical services, changes in general surgical training to include general paediatric surgery or proleptic appointments may be required. PMID- 11918326 TI - Effect of once weekly pulmonary rehabilitation on exercise tolerance in patients with chronic lung disease. AB - BACKGROUND: Pulmonary rehabilitation (PR) programmes improve exercise tolerance and reduce breathlessness in patients with chronic obstructive pulmonary disease. AIM: A retrospective audit was undertaken to assess the effectiveness of the pulmonary rehabilitation programme at Belfast City Hospital (BCH). METHODS: Data from 74 patients who completed a six-week PR programme were reviewed. Exercise tolerance was assessed by the shuttle walk test (SWT), and breathlessness by the BORG scale. Paired t-tests were used for within group analysis. RESULTS: The mean (SD) exercise tolerance assessed by the SWT improved significantly as did duration of individual exercises. Patients were able to exercise more without increasing their breathlessness or heart rate. CONCLUSIONS: This paper highlights the effect of PR on exercise tolerance and breathlessness in patients with chronic lung disease. The results compare favourably with published literature where more frequently supervised exercise sessions were provided. PMID- 11918327 TI - Value of sentinel node biopsy in the management of breast cancer. AB - AIMS: To determine the rate of detection of the sentinel node using both blue dye and radioisotope, and the accuracy with which the sentinel node histology reflects the nodal status of the axilla in a series of patients with clinically node-negative breast cancer. PATIENTS AND METHODS: During a 32-month period from May 1998 to December 2000, 73 patients with clinically node-negative breast cancer underwent sentinel node biopsy immediately followed by formal axillary lymphadenectomy. The sentinel node(s) was identified using a combination of lymphoscintigraphy, blue dye and an intraoperative hand-held gamma probe. RESULTS: The mean age of the 73 patients was 58 years (range 32-83 years). Twenty six per cent (19/73) had previous surgical/excisional biopsy. Pre-operative lymphoscintigraphy was positive in 74% (54/73) of patients. Combination of blue dye and radioisotope was better than either method in isolation for identifying the sentinel node, yielding a success rate of 96% (70/73). A total of 32 cases proved to have positive nodal disease on histological examination. In 44% (14/32) of patients, the sentinel node was the only positive node. Forty-seven per cent (15/32) of patients in whom the sentinel node was positive also had positive nodes in the axillary nodal basin. There were 3/32 false negative cases, giving a false negative rate of 9.4%. CONCLUSION: Sentinel node biopsy will have a role in the management of breast cancer. However, widespread adaptation of this technique awaits the results of prospective, randomised trials. PMID- 11918329 TI - Carers' views on services for the elderly mentally ill. AB - BACKGROUND: Recent health policy emphasises the involvement of the consumer and carer in health and social care issues. Demographic projections predict a rise in the number of elderly people in Ireland and the number of elderly people with a mental illness. The identification of gaps in service provision should result in appropriate planning for this group in the future. OBJECTIVE: The aim of the study was to assess carers' opinions of services for the elderly mentally ill in the Limerick area. METHOD: A questionnaire was sent to the carers of patients referred to the Psychiatry of Old Age service. Information was gathered on satisfaction with existing services for the elderly. Carers were also asked to identify which services they felt they needed. RESULTS: The overall response rate was 51%, but the response rate for individual items varied. Both formal and informal services for the elderly are underused. The need for long-term care places was highlighted and carers wished for more information and emotional support. CONCLUSION: This study highlighted the aspects of the service that carers value, which should help guide future service development. PMID- 11918328 TI - Safety guidelines for radiolocalised sentinel node resection. AB - BACKGROUND: Sentinel node radiolocalisation procedures are associated with low levels of radiation exposure. Radioactive material is present in the operating theatre and pathology laboratory. In most hospitals there are no official regulations in place for sentinel node radiation exposure. AIM: To establish guidelines on the safety of sentinel node mapping with emphasis on the management of radioisotopes. METHODS: The current literature regarding sentinel node procedures and radiation safety was reviewed. EU and US radiation safety regulations were scrutinised. RESULTS: Personnel involved in sentinel node procedures are exposed to low levels of radiation. These levels are not high enough to require designated radiation workers in the theatre and pathology laboratory. Awareness of radiation safety and certain precautions during the procedure and processing of the specimen can further reduce levels of exposure. CONCLUSION: Although low levels of radiation exposure are associated with sentinel node procedures, awareness of radiation safety and adherence to regulations, along with close interdepartmental co-operation, are recommended for further reduction in radiation exposure and safe application of this technique. PMID- 11918330 TI - The sinister significance of dysphagia. AB - BACKGROUND: The majority of patients presenting with oesophageal cancer have symptoms for more than three months and advanced disease at presentation. Most appear unaware of the significance of dysphagia as a symptom. Cancer awareness programmes focus on symptoms such as lumps and bleeding. AIM: To sample the level of public awareness of the potentially sinister significance of the symptom of dysphagia. METHODS: A community survey was conducted using a questionnaire to evaluate the subjects' impression of the significance of dysphagia, and compare it with their perception of the significance of breast lump. Patients were stratified to male and female, under and over 45 years. RESULTS: There were 164 subjects interviewed. Seventy-five per cent stated that they would visit their doctor within one week of developing dysphagia compared with 87 per cent questioned about a breast lump (96 per cent females, 80 per cent males). Only 17 per cent felt that cancer was a probable explanation for dysphagia compared with 80 per cent who would consider cancer a likely cause of breast lump. CONCLUSION: There is evident need of an awareness programme of the potential significance of dysphagia if prognosis for oesophageal cancer is to be improved. PMID- 11918332 TI - Hepatic abscesses in elderly patients mimicking metastatic disease. AB - BACKGROUND: There is considerable overlap between the clinical presentation and radiological appearances of hepatic abscesses and hepatic metastases. The distinction is important given the treatable nature of hepatic abscesses compared with most forms of metastatic disease and the very high morbidity and mortality associated with untreated or missed pyogenic abscesses. AIMS: The aim of this series of case reports is to illustrate this point by presenting the case histories of three elderly patients whose clinical and radiological findings suggested metastatic liver disease, but who were subsequently proven to have liver abscesses. METHODS: A comprehensive review of the clinical and radiological records of three patients. RESULTS: Ultrasound and computer tomography (CT) imaging in all three cases was suggestive of metastatic liver disease. The liver lesions were subsequently proven to be abscesses either by autopsy, needle aspiration or inspection at open surgery. CONCLUSIONS: Liver abscesses can mimic metastatic deposits. Correlation with the white cell count (WCC) can be very helpful. Fine needle aspiration (FNA) of liver lesions should be undertaken, especially if the WCC is elevated. PMID- 11918331 TI - Molecular genotyping of human cryptosporidiosis in Northern Ireland: epidemiological aspects and review. AB - BACKGROUND: Cryptosporidium parvum is the most common of the protozoal pathogens associated with gastrointestinal disease in Northern Ireland. Genotyping techniques are valuable in helping to elucidate sources and modes of transmission of this parasite. There have been no reports on the prevalence of genotypes in Northern Ireland, mainly due to a lack of discriminatory genotyping techniques, which recently have become available. AIM: To investigate the genotype of C. parvum oocysts isolated from human faeces in sporadic cases of cryptosporidiosis in Northern Ireland. METHODS: Thirty-nine isolates of C. parvum, representing 79.6% of the total 1998 laboratory reports for the Eastern Health and Social Services Board, were investigated. Following DNA extraction from oocysts the thrombospondin-related adhesive protein 2 (TRAP-C2) locus was amplified by polymerase chain reaction (PCR) and subsequently sequenced. RESULTS: The majority of isolates (87.2%) were classified as bovine genotype II with the remainder (12.8%) being the human genotype I. CONCLUSIONS: There is a high prevalence of the bovine genotype II parasite in sporadic cases around the greater Belfast area. Epidemiologically, this suggests that the most frequent mode of transmission may be from animals to humans, but does not suggest a high proportion of human to human spread. PMID- 11918333 TI - Clinical elective at Monze, Zambia. PMID- 11918334 TI - Irish medical historiography. PMID- 11918335 TI - Emmanuel Ciprian Amoroso. PMID- 11918336 TI - Revision of a 30-year-old ring total hip prosthesis. PMID- 11918337 TI - Palliation of malignant dysphagia. PMID- 11918338 TI - Major orthopaedic trauma in Jehovah's Witnesses. PMID- 11918339 TI - Aggressive angiomyxoma of the perineum. PMID- 11918340 TI - Spatial infectious disease surveillance in New Zealand: promoting excellence through the application of technology. PMID- 11918341 TI - Comments on case report--self-castration. PMID- 11918342 TI - Assessing dietary intake in the management of obesity. AB - This paper focuses on assessing the caloric-intake side of the energy balance equation in clinical settings. In the treatment of obesity, dietary assessment may have many purposes including the following: 1) establishing a baseline of eating patterns to determine targets of intervention and to gauge progress, 2) providing a means of monitoring change in the targeted dietary areas and behaviors, and 3) allowing for ongoing feedback to the patient. The types of data to be gathered in dietary assessment will depend on the purposes of the assessment. The nature, advantages, and disadvantages of the following dietary assessment methods are reviewed: 24-hour recall, diet history interview and questionnaire, and self-monitoring. When used on an ongoing basis in treatment, self-monitoring enhances weight-loss outcomes. However, compliance with self monitoring varies widely across patients and over time. Possible methods of increasing compliance are discussed. Recent technological advances in software and hardware systems offer promise in improving compliance and effectiveness of self-monitoring. PMID- 11918343 TI - Estimating resting energy expenditure in obesity. AB - In the treatment of obesity, assessment of resting energy expenditure (REE) can provide the basis for prescribing an individualized energy intake to attain a desired level of energy deficit. Indirect calorimetry is the most frequently used method to measure REE, but the great expense of equipment precludes its widespread use. As a result, REE is often estimated by predictive formulas based on weight, height, age, and gender. This paper examines the accuracy of these formulas in estimating REE among obese patients, discusses the assumptions underlying their use, and reviews the need for technological advances that will make the assessment of REE accurate, portable, and inexpensive. PMID- 11918344 TI - HIV fusion inhibitors for AIDS therapy. PMID- 11918345 TI - Chemokine receptors--the next therapeutic target for HIV? AB - To date, the available therapies for the treatment of HIV infection are targeted against proteins encoded by the virus itself. Thus, combination drug therapies for HIV with reverse transciptase and protease inhibitors have resulted in spectacular reductions of viraemia, often leading to a remarkable improvement in symptoms and recovery from disease in infected people. There is still however, a great need for improved therapies since many patients are unable to take these drugs, either for reasons of intolerance, strain resistance, complexity of regimen or prohibitive cost. Multiple therapies aimed at different events in the HIV life cycle will ensure switching of treatments to combat resistant viruses, and also allow treatment flexibility if patients are unable to tolerate particular therapies. One event that could provide a key to reducing or even eliminating viral infection would be to prevent the virus from entering the host cell. Intense efforts are now underway to produce drugs that target chemokine receptors, one of the essential components for HIV cell entry. HIV needs two receptors on the host cell surface for efficient attachment and infection. The first is CD4 and the second, identified in 1996, is a member of the family of chemokine receptors, members of the G-protein coupled 7TM superfamily, which are involved in the trafficking of leukocytes in immune surveillance and inflammation. Many small, orally bioavailable molecules that block various 7TM receptors are used to treat a panoply of diseases including ulcers, allergies, migraines, and schizophrenia. These molecules are the cornerstone of the pharmaceutical industry's contribution to the fight against so many diseases. Small molecule inhibitors of the HIV-coreceptors are now entering the first stages of clinical trials as new therapeutics for the fight against AIDS. PMID- 11918347 TI - Dopamine and epinephrine, but not serotonin, downregulate dopamine sensitivity in cultured cortical and striatal astroglial cells. AB - Biogenic amines are important in the regulation of neuronal functions and complex behavior in the brain. However, putative contributions of glial cells to physiological effects of aminergic transmitters and their pathophysiological implications are poorly understood. Astrocytes are known to respond to dopamine (DA) with calcium transients that can be blocked by the D1- and D2-receptor subtype specific antagonists SCH23390 and Sulpiride. We demonstrate here that DA sensitivity of cortical and striatal astrocytes is changed by application of either DA or epinephrine (EP), but not serotonin (5-HT). Exposure of cortical and striatal astroglial cultures for less than 1h to DA (> or = 10 micromol/l) or EP (> or = 1 micromol/l) leads to a significant reduction of astroglial DA sensitivity. Whereas the DA-mediated downregulation of astroglial DA-sensitivity can be reverted by SCH23390 (> or = 1 micromol/l), and Sulpiride (> or = 10 micromol/l), EP-mediated effects are insensitive to these antagonists. In contrast to receptor function, expression of D1- and D2-DA-receptors is not altered by either DA or EP in cortical and striatal astroglial cells as revealed by western blot analysis. Our results demonstrate that sensitivity of astroglial cells to DA is modulated by DA and EP, adding new evidence to a role of astrocytes as targets for physiological and pathological effects of aminergic transmitters. PMID- 11918346 TI - 2-Aminoethoxydiphenyl borate (2-APB) inhibits capacitative calcium entry independently of the function of inositol 1,4,5-trisphosphate receptors. AB - Capacitative calcium entry (CCE), the mechanism that replenishes intracellular calcium stores after depletion, is essential to intracellular calcium signaling. CCE is mediated by the channels in the plasma membrane generally referred to as "store operated channels (SOCs)". However, the molecular identity of the SOCs has never been determined, and the mechanism of the activation of SOCs remains to be elucidated. Recent studies have demonstrated that 2-aminoethoxydiphenyl borate (2 APB), which has been found to be an antagonist of inositol 1,4,5-trisphosphate receptors (IP3Rs), inhibits CCE, suggesting that IP3Rs channel activity is essential to the generation of CCE. However, CCE has also been reported to occur normally in IP3R-deficient cells. In order to resolve this discrepancy, we investigated the effect of 2-APB on CCE in IP3Rs-deficient cells. In response to store depletion with thapsigargin or N,N,N',N'-tetrakis (2-pyridylmethyl) ethylene diamine (TPEN), CCE was generated in IP3Rs-deficient cells the same as in wild-type cells, however, 2-APB abolished CCE in IP3Rs-deficient cells, despite the fact that this cell line does not possess functional IP3Rs. We also examined the effect of 2-APB on several types of TRP Ca2+ channels, which exhibit properties similar to those of SOCs. 2-APB had a different inhibitory effect on spontaneous and thapsigargin-induced Ba2+ influx in cells that transiently expressed individual TRP subtypes. These results suggest that the channel activity of IP3Rs is not essential to the generation of CCE in this cell line and that 2-APB inhibits CCE independently of the function of IP3Rs. PMID- 11918348 TI - Solubilization and purification of the human ETB endothelin receptor produced by high-level fermentation in Pichia pastoris. AB - In the present report, the successful solubilization and purification of the ETB receptor heterologously produced in the methylotrophic yeast P. pastoris is described for the first time. In comparison to the baculovirus system where successful production, solubilization and purification have already been reported, handling and up-scaling of recombinant P. pastoris cells was much easier and less time consuming. Recombinant P. pastoris clones producing two different ETB receptor constructs were grown in a fermenter to a density of about 360 g/l. After induction with methanol, a production level of maximally 45 pmol/mg was obtained, a value which is in the range of that reported for baculovirus-infected insect cells. A method for the large-scale preparation of membranes was established. Solubilization of the recombinant ETB receptor was achieved with the detergent n-dodecyl-/beta-D-maltopyranoside. The stability of the solubilized and ligand-bound receptor was examined in detail. Subsequently, two purification methods for two different receptor constructs were tested and a large-scale procedure for isolation of recombinant receptor was established. In general, the purification methods described herein will be adaptable to other G protein-coupled receptors heterologously produced in heterologous expression systems including P. pastoris. PMID- 11918349 TI - Rat brain cholinergic, dopaminergic, noradrenergic and serotonergic neurons express GABAA receptors derived from the alpha3 subunit. AB - In order to study the most abundant GABAA receptor subtypes expressed in cholinergic, dopaminergic, noradrenergic and serotonergic neurons (i.e., in neurons of the so-called "global" projection systems), we employed double immunocytochemical techniques combining the labeling of GABAA receptor alpha1, alpha2 and alpha3 subunit with markers for these cells. Cholinergic neurons in the striatum, habenula, and pedunculo-pontine nucleus were immunonegative for the alpha1 subunit, and most were also alpha2-immunonegative. However, cholinergic neurons in the striatum, septum and pedunculo-pontine nucleus were alpha3 immunopositive. Dopaminergic neurons in the substantia nigra pars compacta were highly immunopositive for the alpha3, and noradrenergic neurons in the locus coeruleus were immunoreactive for the alpha3 and the alpha2-subunit; although neurons of these areas were negative for alpha1. Similarly, serotonergic neurons in raphe also showed a high level of labeling of alpha3, while there was a lack of immunoreactivity for the alpha1-subunit, and only some individual neurons were positive for the alpha2 subunit. As the presence of different alpha-subunits confers specific physiological and pharmacological properties to GABAA receptors, the abundance of receptors containing the alpha3 subunit (and the scarcity of receptor subtypes including the other alpha-subunits studied) may have important implications for the GABAergic regulation of brain "global" or "diffuse" projection systems. PMID- 11918350 TI - Patterns of cyclic AMP formation by coexpressed D1 and D2L dopamine receptors in HEK 293 cells. AB - Recent studies using immunofluorescence confocal microscopy (Aizman et al., Nature neuroscience (2000) 3, 226-230) present compelling evidence for colocalization of D1 and D2 dopamine receptors on neurons in the striatum and nucleus accumbens. To examine some of the biochemical consequences of colocalization we coexpressed the D1 and D2 dopamine receptors in HEK293 cells. Dopamine D1 and D2 receptors couple to stimulation and inhibition of adenylyl cyclase, respectively. In cells expressing only the D1 receptor, dopamine stimulated cAMP formation with an EC50 of 2.15 nM. In cells expressing only the D2L receptor, dopamine inhibited cAMP formation by 80% with an EC50 of 0.02 nM. The effect of dopamine on the D2L receptor was antagonized by the selective antagonist spiperone with an IC50 of 0.31 nM. In cells coexpressing both the D1 and D2L receptors, dopamine caused an increase in cAMP that was only 20% of that observed with the D1 receptor alone. In this case, increasing concentrations of spiperone caused a change in the dose-response curve from hyperbolic to bell shaped as the concentration of spiperone was increased. Using pharmacological constants determined from studies on the individually expressed receptors, the curves obtained in cells co-expressing the two receptors could be modeled by kinetic expressions derived by summing the contributions from each receptor. The model leads to a re-interpretation of the pharmacology of dopaminergic ligands. Hence, one consequence of colocalization is that D2 receptor antagonists become functional agonists of cAMP formation. PMID- 11918351 TI - Neuromuscular transmission on the rebound. AB - Recent work at the zebrafish neuromuscular junction (NMJ) has shown that positively charged acetylcholine (ACh), at the high concentrations reached in the cleft during neuromuscular transmission, blocks acetylcholine receptors (AChRs) as soon as they open. Thus after two ACh molecules bind and open the channel, a third molecule enters and blocks the pore at a site resembling that for block by local anesthetics, suggesting that ACh is the endogenous anesthetic of the NMJ. Recovery from open channel block results in a rebound synaptic current only after ACh is cleared from the cleft. Kinetic modeling of other AChRs suggests that a rebound current is generated at all vertebrate NMJs, from fish to frogs to mammals. Open channel block prolongs the current at fast zebrafish NMJs in order to more effectively spread charge along the fibers, akin to multiple central synapses spread over dendrites. Together these findings indicate the need for a fundamental revision of current thinking about neuromuscular transmission at many levels, including channel structure, function and pharmacology. PMID- 11918352 TI - Computer-aided diagnosis in radiology. PMID- 11918353 TI - The NCI Biomedical Imaging Program: five-year progress report. PMID- 11918354 TI - Use of an artificial neural network to determine the diagnostic value of specific clinical and radiologic parameters in the diagnosis of interstitial lung disease on chest radiographs. AB - RATIONALE AND OBJECTIVES: The authors investigated the diagnostic value of each of multiple clinical parameters and radiologic findings in differentiating between various interstitial lung diseases by using an artificial neural network (ANN). MATERIALS AND METHODS: The ANN was designed to differentiate between 11 interstitial lung diseases. The authors employed 10 clinical parameters and 16 radiologic findings that were divided into three groups (location, general appearance, specific findings). The performance of the ANN was evaluated with receiver operating characteristic analysis with amodified round-robin (leave-one out) method and 370 cases (150 actual cases, 110 published cases, and 110 hypothetical cases). The Az values of ANNs were evaluated with various combinations of 10 clinical parameters and 16 radiologic findings. RESULTS: The Az value obtained with the complete set of clinical parameters and radiologic findings was 0.947. The Az value obtained with the 10 clinical parameters alone was 0.900, which was greater than 0.843 obtained with the 16 radiologic findings alone. There were statistically significant differences among Az values for some diseases when certain clinical parameters were removed from the input. Omission of specific findings among the three groups of radiologic findings decreased the Az value significantly. CONCLUSION: These results appear to confirm that clinical parameters can be equally as or more important than radiologic findings in the diagnosis of interstitial lung diseases. Among radiologic findings, certain specific findings can be more important than the location or general appearance of abnormal findings. PMID- 11918355 TI - Can the size of microcalcifications predict malignancy of clusters at mammography? AB - RATIONALE AND OBJECTIVES: The purpose of this study was to determine whether the size of mammographically detected microcalcifications is predictive of malignancy. MATERIALS AND METHODS: Two hundred sixty mammograms showing clustered microcalcifications with proven diagnoses (160 malignant, 100 benign) were respectively reviewed by experienced mammographers. Lesions that were obviously benign in appearance were excluded from the study. A computer-aided diagnosis system digitized the lesions at 600 dpi, and the microcalcifications on the digital image were interactively defined by mammographers. Subsequently, three quantitative features that reflected the size of the microcalcifications-length, area, and brightness-were automatically extracted by the system. For each feature, the standard average of values obtained for individual calcifications within the cluster and the average with emphasis on extreme values (E) obtained in a single cluster were analyzed and matched with pathologic results. RESULTS: In the malignant group of cases, the mean values of the standard average length and area were significantly higher (P < .0001) than the mean values in the benign group. Distribution analysis demonstrated that an average length of more than 0.41 mm was associated with malignant lesions 77% of the time, while an average length of less than 0.41 mm was associated with benign lesions 71% of the time. The mean of the average length (E) and area (E) of microcalcifications within the cluster demonstrated an even higher discriminative power when compared with the standard average length and area. The average brightness, on the other hand, showed only a low discriminative power. CONCLUSION: Digital computerized analysis of mammographically detected calcifications demonstrated that the average length and area of the calcifications in benign clusters were significantly smaller than those in malignant clusters. PMID- 11918356 TI - Factor analysis of medical image sequences improves evaluation of first-pass MR imaging acquisitions for myocardial perfusion. AB - RATIONALE AND OBJECTIVES: Factor analysis of medical image sequences (FAMIS) applied to gadolinium chelate-enhanced subsecond magnetic resonance (MR) imaging was evaluated as a postprocessing method for assessing myocardial perfusion in coronary artery disease (CAD). MATERIALS AND METHODS: To assess the accuracy of motion correction, five normal volunteers underwent MR imaging at rest. Thirteen patients with well-documented CAD and no myocardial infarction underwent MR imaging at rest and after dipyridamole administration. After motion correction, a single myocardial tissue factor (FAMISt) image was obtained with FAMIS for each raw MR imaging series acquisition. To evaluate how FAMIS could improve the analysis of these acquisitions, five readers visually assessed myocardial perfusion with FAMISt and raw MR images, and a multicase, multireader receiver operating characteristic analysis was performed. RESULTS: FAMISt images significantly improved detection of the perfusion defects when compared with raw MR images (P = .002). Areas under the receiver operating characteristic curves ranged from 0.84 to 0.93 with FAMISt images and from 0.48 to 0.85 with raw MR images. CONCLUSION: FAMIS applied to first-pass MR imaging series provided myocardial perfusion images that improve the objective assessment of myocardial perfusion in patients with CAD. PMID- 11918357 TI - Computer-aided diagnosis of masses with full-field digital mammography. AB - RATIONALE AND OBJECTIVES: The authors developed and evaluated a method of computer-aided diagnosis (CAD) for mass detection with full-field digital mammography (FFDM). MATERIALS AND METHODS: The new CAD method for FFDM employs adaptive, nonlinear multiscale processing and hybrid classification methods. The major strategies are (a) to "standardize" the mammographic image before it is input to the analysis modules, (b) to adapt the segmentation of suspicious regions adapt to accommodate different characteristics of masses and mammograms, and (c) to use combined "hard" and "soft" decision making in discriminating between mass and normal tissue regions. Two data sets of diagnostic FFDM mammograms were used. The training data set includes 36 normal and 24 abnormal mammograms (34 masses), and the testing data set includes 24 normal and 10 abnormal mammograms (10 masses). The tumors in this diagnostic database were more subtle and difficult to detect than those in screening databases the authors have used before. RESULTS: With the limited database and a partial optimization, a sensitivity of 91% was obtained in training, with a false-positive rate of 3.21 per image. At this trained operating point of the CAD system, six of 10 subtle masses were detected in testing. CONCLUSION: The CAD algorithms developed in screen-film mammography can be modified for FFDM. More data analysis and system optimization and evaluation will be needed before CAD can be integrated efficiently into the performance of FFDM. PMID- 11918358 TI - Tutor versus computer: a prospective comparison of interactive tutorial and computer-assisted instruction in radiology education. AB - RATIONALE AND OBJECTIVES: This study compared the educational effectiveness of an interactive tutorial with that of interactive computer-assisted instruction (CAI) and determined the effects of personal preference, learning style, and level of training. MATERIALS AND METHODS: Fifty-four medical students and four radiology residents were prospectively, randomly assigned to receive instruction from different sections of an interactive tutorial and an interactive CAI module. Participants took tests of factual knowledge at the beginning and end of the instruction and a test of visual diagnosis at the end. They completed questionnaires to evaluate their preferred learning styles objectively and to elicit their subjective attitudes toward the two formats. Mean test scores of the tutorial and CAI groups were compared by means of analysis of covariance and two tailed repeated-measures F test. RESULTS: Both the tutorial and CAI groups demonstrated significant improvement in posttest scores (P < .01 and P < .01, respectively) with the tutorial group's mean posttest score marginally but significantly higher (32.84 vs 28.13, P < .001). There were no significant interaction effects with participants' year of training (P = .845), objectively evaluated preferred learning style (P = .312), subjectively elicited attitude toward learning with CAI (P = .703), or visual diagnosis score (tutorial, 7.61; CD-ROM, 7.75; P = .79). CONCLUSION: Interactive tutorial and optimal CAI are both effective instructional formats. The tutorial was marginally but significantly more effective at teaching factual knowledge, an effect unrelated to students' year of training, learning style, or stated enjoyment of CAI. The superiority of the tutorial is expected to increase when it is compared with commercially expedient CAI modules. PMID- 11918359 TI - Gadolinium-enhanced 3D MR angiography of renal artery stenosis: a pilot comparison of maximum intensity projection, multiplanar reformatting, and 3D volume-rendering postprocessing algorithms. AB - RATIONALE AND OBJECTIVES: The authors compared diagnostic accuracy of maximum intensity projection (MIP), multiplanar reformatting (MPR), and three-dimensional (3D) volume rendering (VR) in the evaluation of gadolinium-enhanced 3D magnetic resonance (MR) angiography of the renal arteries. They hypothesized that VR is as accurate as or more accurate than MIP and MPR at depicting renal artery stenosis. MATERIALS AND METHODS: The study group comprised 28 consecutive patients who underwent gadolinium-enhanced 3D MR angiography of the renal arteries. Studies were postprocessed to display images in MIP, MPR, and VR formats. Digital subtraction angiography (DSA), when performed (nine of 28 patients), was the standard for comparison. For each main renal artery, an estimate of percentage stenosis was made for any stenoses detected by three independent radiologists. For calculation of sensitivity, specificity, and accuracy, MR angiographic stenosis estimates were categorized as mild (0%-39%), moderate (40%-69%), or severe (> or = 70%). DSA stenosis estimates of 70% or greater were considered hemodynamically significant. RESULTS: Analysis of variance demonstrated MIP estimates of stenosis were statistically greater than VR estimates in two readers and greater than MPR estimates in all readers for all patients. MIP images also showed the largest mean difference from DSA stenosis estimates for all three readers. For both VR and MPR, mean differences between MR angiographic stenoses estimates and DSA estimates reached significance for only one reader, whereas, for MIP versus DSA, mean differences reached significance for all three readers. Although not statistically significant compared with DSA, accuracies of VR (87%) and MPR (89%) were greater than that of MIP (81%). CONCLUSION: In this pilot study, MIP was the least accurate of the three image display algorithms tested. VR and MPR yielded similar values for each method of comparison. PMID- 11918360 TI - System for robotically assisted prostate biopsy and therapy with intraoperative CT guidance. AB - RATIONALE AND OBJECTIVES: The purpose of this study was to assess the work-in progress prototype of an image-guided, robotic system for accurate and consistent placement of transperineal needles into the prostate with intraoperative image guidance inside the gantry of a computed tomographic (CT) scanner. MATERIALS AND METHODS: The coach-mounted system consists of a seven-degrees-of-freedom, passive mounting arm: a remote-center-of-motion robot; and a motorized, radiolucent needle-insertion device to deliver 17-18-gauge implant and biopsy needles into the prostate with the transperineal route. The robot is registered to the image space with a stereotactic adapter. The surgeon plans and controls the intervention in the CT scanner room with a desktop computer that receives DICOM images from the CT scanner. The complete system fits in a carry-on suitcase, does not need calibration, and does not utilize vendor-specific features of the CT scanner. RESULTS: In open air, the average accuracy was better than 1 mm at a 5-8 cm depth. In various phantoms, the average orientation error was 1.3 degrees, and the average distance between the needle tip and the target was 2 mm. CONCLUSION: Results of preliminary experiments indicate that this robotic system may be suitable for transperineal needle placement into the prostate and shows potential in a variety of other percutaneous clinical applications. PMID- 11918361 TI - Factors that affect National Resident Matching Program rankings of medical students applying for radiology residency. AB - RATIONALE AND OBJECTIVES: A study was performed to determine which factors most influence the program rank lists of U.S. medical students applying for radiology residency and which sources of information about residency programs these medical students value most highly. MATERIALS AND METHODS: Questionnaires were mailed to the 516 U.S. medical students who applied to the authors' radiology residency program in the 2001 National Resident Matching Program match. Respondents were asked to evaluate 23 factors that may have affected their rank lists, on a five point Likert scale. For data interpretation, factors were categorized as being either within, somewhat within, or outside the control of the program director. Respondents were also asked to evaluate eight potential sources of information about residency programs. RESULTS: The response rate was 23.3% (120 of 516). The three most important factors affecting rank lists were the perceived happiness of current residents within the program, the geographic location of the program, and the academic reputation of the department. Four of the five factors that scored most highly were judged to be outside the program director's control. The most important sources of information were the interview day and personal interactions with residents currently in the program. Program Web sites were rated more highly than the written materials provided. CONCLUSION: Medical students applying for radiology residency use many sources of information and weigh many factors in making their program rank lists. Many of the most influential factors are outside the direct control of the program director. PMID- 11918362 TI - Retention of radiographic anatomy of the chest by 4th-year medical students. AB - RATIONALE AND OBJECTIVES: The authors performed this study to assess the ability of medical students to retain their knowledge of radiographic anatomy into the 4th year. MATERIALS AND METHODS: One hundred ninety-four 4th-year medical students were quizzed on their ability to (a) identify 10 anatomic structures on chest radiographs and (b) search an abnormal radiograph to identify two important findings. Frontal and lateral views were used. The students had learned about radiographic anatomy of the chest and plain radiography search techniques in 1st- and 2nd-year courses and had proved their ability to perform these tasks, during their 2nd year of medical school, by achieving grades of 84% or more (average grade, 90%) on individual oral examinations administered by radiologists. RESULTS: The mean test score was 4.47 out of 12. Only 10 students (5%) were able to identify the main pulmonary artery on the frontal view, and only nine (4%) identified the main bronchi on the lateral view. Of the 194 students, 192 failed to detect a collapsed vertebral body on the lateral view of a patient who had previously undergone a mastectomy. CONCLUSION: Specific radiologic anatomy facts and search techniques are poorly retained by 4th-year medical students. Preclinical teaching of imaging probably should stress principles rather than specific details. PMID- 11918363 TI - Results of the 2001 survey of the American Association of Academic Chief Residents in Radiology. AB - RATIONALE AND OBJECTIVES: The American Association of Academic Chief Residents in Radiology (A3CR2) conducts an annual survey of residency training programs. The survey data allow comparative analyses to be performed among training programs regarding resident education, benefits, clinical demands, and other resident related issues. MATERIALS AND METHODS: Questionnaires were mailed to all accredited programs registered in the A3CR2 database (150 total programs). The yearly questions address demographic information concerning the individual programs including academic affiliation, number of radiologic examinations performed per resident, changes in the number of residents per program, and financial compensation. The rotating questions, which are revisited every 4 years, focused on preparation for the American Board of Radiology examinations, educational issues, and employment outlook. RESULTS: Completed surveys were received from 55 programs (37.3%). Among the responses, three issues prevailed: (a) increased number of examinations performed per resident, with resultant decrease in educational time, (b) decreased quality of education in all sections of radiology and decreased number of educational conferences, and (c) improved employment outlook, but continued trend of fewer residents choosing a career in academics. CONCLUSION: The current boom in the radiology job market and the increased number of radiologic examinations performed annually appear to adversely affect radiologic education through a decreased number of conferences and an increased number of radiologic examinations performed per resident. PMID- 11918364 TI - Effects of ioxaglate on cultured microvascular endothelial cells: do all in vitro studies actually reflect clinical situations? PMID- 11918365 TI - Functional CT imaging of the lungs: the pulmonary function test of the new millennium? PMID- 11918366 TI - Subsecond multisection CT of regional pulmonary ventilation. AB - RATIONALE AND OBJECTIVES: To evaluate the adequacy of multibreath and single breath stable xenon gas techniques to measure regional ventilation during cardiac gated, high-speed, multisection imaging, the authors carried out a series of studies using electron-beam computed tomography (CT) and a recently introduced subsecond multisection spiral CT scanner. MATERIALS AND METHODS: In four anesthetized pigs, the authors implemented single-breath and/or dynamic multibreath wash-in and washout protocols with respiratory-- and cardiac-gated image acquisition. The effects of varying tidal volume and inspiratory flow rate were evaluated independently. Scanning was done at end expiration to avoid artifacts from partial volumed conducting airways, which are filled with inspired gas concentration during inspiration. RESULTS: A single breath of 100% xenon provides adequate enhancement in the lung parenchyma (mean, 32 HU +/- 1.85 [standard error]) and should not cause unwanted side effects (mean xenon concentration in lung periphery, 21%). The single-breath method is suitable for studies requiring only short periods of apnea. Using the multibreath method, in dependent portions of the lung, there was close agreement between measured changes and predictions based on the xenon calibration data. More than 10 breaths were needed to clear tracer from poorly ventilated areas, and some nondependent regions demonstrated apparently "linear" rather than exponential clearance curves, possibly reflecting longer washout times. Analysis of wash-in and washout curves revealed vertical ventilation gradients and, at higher inspiratory flow rates, redistribution of ventilation to areas of the lung with greater pathway conductance. CONCLUSION: With careful attention to lung volume and use of cardiac gating, it is now possible to correlate lung structure with function to a degree heretofore not possible. PMID- 11918367 TI - Statistical power in observer-performance studies: comparison of the receiver operating characteristic and free-response methods in tasks involving localization. AB - RATIONALE AND OBJECTIVES: Statistical power, defined as the probability of detecting real differences between imaging modalities, determines the cost in terms of readers and cases of conducting receiver operating characteristic (ROC) studies. Neglect of location information in lesion-detection studies analyzed with the ROC method can compromise power. Use of the alternative free-response ROC (AFROC) method, which considers location information, has been discouraged, because it neglects intraimage correlations. The relative statistical powers of the two methods, however, have not been tested. The purpose of this study was to compare the statistical power of ROC and AFROC methods using simulations. MATERIALS AND METHODS: A new model including intraimage correlations was developed to describe the decision variable sampling and to simulate data for ROC and AFROC analyses. Five readers and 200 cases (half of which contained one signal) were simulated for each trial. Two hundred trials, equally split between the null hypothesis and alternative hypothesis, were run. Ratings were analyzed with the Dorfman-Berbaum-Metz method, and separation of the null hypothesis and alternative hypothesis distributions was calculated. RESULTS: The AFROC method yielded higher power than the ROC method. Separation of the null hypothesis and alternative hypothesis distributions was larger by a factor of 1.6 regardless of the presence or absence of intraimage correlations. The effect of the incorrect localizations during ROC analysis of localization data is believed to be the major reason for the enhanced power of the AFROC method. CONCLUSION: The AFROC method can yield higher power than the ROC method for studies involving lesion localization. Greater consideration of this methodology is warranted. PMID- 11918368 TI - Projected digital radiologic images for teaching: balance of image quality with data size constraints. AB - RATIONALE AND OBJECTIVES: The authors performed this study to determine, in the context of a teaching presentation with slides, the information content needed in a digitized radiologic image for it to be equivalent in quality to an analog image. MATERIALS AND METHODS: Eleven sets of radiologic images were obtained from a teaching file collection and digitized. The images were scanned at high resolution and saved as image files. The information content of each image was then halved repeatedly with repeat sampling of the image. This procedure was repeated nine times to yield a total of 10 images with an information content that ranged from very low (32 kB) to high (12 MB). Each of these image files was made into a 35-mm slide by using a digital slide maker. The original radiographs were subsequently made into slides by using conventional photographic methods. Care was taken to make the images identical in every respect except digitization and information content. The slides were shown to radiologists, who filled out questionnaires to rate image quality. RESULTS: Digitized slides that contain an average of 416 kB of data (the equivalent of a 750 x 570-pixel matrix with 8-bit gray levels) were equivalent to analog images for 90% of viewers. At 830 kB (1,060 x 800-pixel matrix with 8-bit gray levels), 100% of viewers found no difference between digitized and analog images. CONCLUSION: Digital images become indistinguishable from analog images if a sufficient amount of information is retained in the digital image. There is a defined relationship between the perception of quality and the information content of a digital image. PMID- 11918369 TI - Imaging findings in intracranial aspergillosis. AB - RATIONALE AND OBJECTIVES: The authors' purpose was to elucidate the various computed tomographic (CT) and magnetic resonance (MR) imaging findings in intracranial aspergillosis. MATERIALS AND METHODS: Retrospective analysis of cranial imaging findings was performed in eight proved cases of central nervous system aspergillosis. The patients ranged in age from 17 to 75 years. Four patients were immunocompromised, and four were immunocompetent. CT was performed in all eight patients, and MR imaging in five. RESULTS: Six patients (75%) had multiple lesions seen on the imaging studies, with a total of 27 focal brain lesions demonstrated. The lesions were most commonly seen in the cerebral hemispheres (n = 21), with lesser involvement of the basal ganglia (n = 2) and the posterior fossa (n = 4). Seven lesions were hemorrhagic on CT and/or MR images. There was a correlation between lesion size and hemorrhage, with hemorrhage more likely in larger lesions (>15 mm). At pathologic examination, foci of hemorrhage were noted within both infarcts and abscesses. Enhancement was noted in five lesions, four of which were confirmed abscesses. Contrast enhancement of the lesions was vague and week in immunocompromised patients but solid and strong in immunocompetent patients. There were 18 lesions without hemorrhage or enhancement; they were either infarcts or abscesses at pathologic examination. Some of these small nonhemorrhagic nonenhancing brain lesions in the subcortical white matter mimicked lacunar infarcts. CONCLUSION: Typical imaging findings of intracranial aspergillosis include multifocal lesions involving the cerebral hemispheres, with hemorrhage in approximately 25% of lesions. Lesional contrast enhancement tends to be stronger in immunocompetent hosts. PMID- 11918370 TI - Validation of injection parameters for catheter-directed intraarterial gadolinium enhanced MR angiography. AB - RATIONALE AND OBJECTIVES: Catheter-directed intraarterial (IA) injections of gadolinium contrast agents may be used during endovascular interventions with magnetic resonance (MR) imaging guidance. Injection protocols require further validation. Using a flow phantom and swine, the authors aimed to (a) measure the optimal arterial gadolinium concentration ([Gd]) required for MR angiography and (b) validate a proposed IA injection protocol for gadolinium-enhanced MR angiography. MATERIALS AND METHODS: For in vitro experiments, the authors placed a catheter in the aorta of an aorto-renal-iliac flow phantom. Injected [Gd], injection rates, and aortic blood flow rates were varied independently for 36 separate IA gadolinium injections. The authors performed 2D and 3D MR angiography with a fast spoiled gradient-recalled echo sequence. For subsequent in vivo experiments, they selectively placed catheters within the aorta, renal artery, or common iliac artery of three pigs. Injection rate and injected [Gd] were varied. The authors performed 32 separate IA gadolinium injections for 2D MR angiography. Signal-to-noise ratios (SNRs) were compared for the various combinations of injection rate and injected [Gd]. RESULTS: In vitro, an arterial [Gd] of 2%-4% produced an optimal SNR for 2D MR angiography, and 3%-5% was best for 3D MR angiography. In swine, an arterial [Gd] of 1%-4% produced an optimal SNR. In the phantom and swine experiments, SNR was maintained at higher injection rates by inversely varying the injected [Gd]. CONCLUSION: Dilute arterial [Gd] is required for optimal IA gadolinium-enhanced MR angiography. To maintain an optimal SNR, injection rates and injected [Gd] should be varied inversely. The postulated injection protocol was validated. PMID- 11918372 TI - Subsidizing radiology research. PMID- 11918371 TI - Near-infrared optical imaging of the breast with model-based reconstruction. AB - RATIONALE AND OBJECTIVES: Near-infrared diffuse optical imaging may offer enhanced contrast resolution over that of the existing technologies for detection and diagnosis of breast cancer. The authors report quantitative absorption and scattering images of the human breast with model-based reconstruction methods using near-infrared continuous-wave tomographic data. MATERIALS AND METHODS: An automatic, multichannel optical imaging system was used to image the breasts of nine women: four with infiltrating ductal carcinomas, one with infiltrating lobular carcinoma, one with fibroadenoma, and three control subjects with no breast lesion. The image reconstruction methods are centered on the finite element solution of photon diffusion in breast tissue. RESULTS: Substantial contrast between tumor and adjacent parenchyma was observed. Images of the control subjects showed homogeneous optical features. In the six women with breast lesions, the locations and sizes of tumors imaged optically were accurate and consistent with the mammographic findings. CONCLUSION: The results of this pilot study show that cancers as small as 5 mm can be quantitatively imaged. In addition, preliminary data from the scattering images suggest that benign and malignant tumors can be noninvasively differentiated with optical imaging. PMID- 11918373 TI - Heating potential of iron oxides for therapeutic purposes in interventional radiology. AB - RATIONALE AND OBJECTIVES: In addition to their diagnostic applications, iron oxides could be used therapeutically to eliminate tumors with heat if their heating powers are adequate. The authors therefore examined the specific absorption rate (SAR) of different iron oxide (magnetite) samples suspended in water and in liquid or solidified gel. MATERIALS AND METHODS: The authors compared two ferromagnetic fine powders (total particle size, >350 nm and 100 nm), five superparamagnetic ferrofluidic samples (total particle size, 10-280 nm), and a commercially available contrast medium (ferumoxides injectable solution, Endorem). The SARs of the magnetic material-suspended in distilled water or in liquid or solid agar-were estimated from time-dependent calorimetric measurements during exposure to an alternating current magnetic field (amplitude, 6.5 kA/m; frequency, 400 kHz). RESULTS: SARs varied considerably between the different iron oxide samples. The highest value was found for a ferrofluidic sample (>93 W/g), while Endorem had little heating power (<0.1 W/g). The SAR was clearly dependent on the aggregation state of the matrix only for the large particle-size ferromagnetic sample, yielding the highest values for particle suspensions in water (74 W/g) and lowest for solid agar (8 W/g). The heating power of the smaller-particle-size ferromagnetic sample did not exceed 8 W/g. CONCLUSION: Heating powers differed according to the interaction of multiple physical parameters. Iron oxides should be selected carefully for therapeutic applications in magnetic heating. PMID- 11918374 TI - Legal regulations on radiation safety for radiologists in Turkey. PMID- 11918375 TI - Relative value of analog and digital teaching files. AB - RATIONALE AND OBJECTIVES: This study was performed to determine if it is financially reasonable for radiology residency programs that create and maintain their own teaching files to switch from analog teaching files (ATFs) to digital teaching files (DTFs). MATERIALS AND METHODS: Radiology residency program directors were surveyed electronically about the monetary value and importance of conventional ATFs and DTFs. The costs for maintaining each type of file were calculated at the authors' institution. RESULTS: Surveys were sent to the program directors of all 197 accredited radiology residencies. Responses were received from 48 (24%). DTFs were scored as more important than ATFs, but the difference was not significant (P = .22). DTFs were rated as less complete (P = .01) but more current (P << .001) than ATFs. DTFs included the American College of Radiology Learning File (85%), in-house productions (77%), and other commercially available products (63%). Thirty percent of respondents had a DTF integrated into a picture archiving and communication system, and 28% reported having a technician dedicated to the teaching file. Program directors ascribed total median dollar values of $250 and $3,000 per year to their ATFs and DTFs, respectively. The annual costs at the authors' institution were much higher than these ascribed values: $44,720 ($91 per case) for maintaining a DTF produced in house and $24,601 ($50 per case) for maintaining an ATF, excluding physician time. CONCLUSION: Program directors are more willing to pay for a DTF than an ATF. For both, the costs of maintenance are great and the relative monetary value is low. PMID- 11918377 TI - Importance of radiology clerkships in teaching medical students life-threatening abnormalities on conventional chest radiographs. AB - RATIONALE AND OBJECTIVES: The purpose of this study was to investigate the ability of medical students who had already completed medicine and surgery clerkships to identify life-threatening abnormalities on conventional chest radiographs. MATERIALS AND METHODS: From May 2000 to August 2001, 72 consecutive 3rd- and 4th-year medical students enrolled in the radiology clerkship at Rhode Island Hospital completed examinations before and after the radiology clerkship. During the examination, they were asked to identify acute abnormalities on five conventional chest radiographs. Only students who had already completed both medicine and surgery clerkships were included in this prospective study. RESULTS: Before the radiology clerkship, the frequency of correct diagnoses of pneumothorax, pneumoperitoneum, congestive heart failure, misplaced endotracheal tube, and misplaced feeding tube were 40%, 61%, 57%, 6%, and 6%, respectively. After the radiology clerkship, these findings were correctly identified more than 94% of the time (P < .001). CONCLUSION: Learning to identify life-threatening abnormalities on conventional chest radiographs through medicine and surgery clerkships is insufficient. The radiology clerkship provides a unique educational experience that significantly improves these abilities. PMID- 11918376 TI - Is past academic productivity predictive of radiology resident academic productivity? AB - RATIONALE AND OBJECTIVES: The authors performed this study to determine whether academic productivity in college and medical school is predictive of the number of publications produced during radiology residency. MATERIALS AND METHODS: The authors reviewed the records of 73 radiology residents who completed their residency from 1990 to 2000. Academic productivity during college, medical school, and radiology residency, other postgraduate degrees, and past careers other than radiology were tabulated. The personal essay attached to the residency application was reviewed for any stated academic interest. Residents were classified as being either previously productive or previously unproductive. Publication rates during residency and immediately after residency were compared for the two groups. For the productive residents, a correlation analysis was used to examine the relationship between past frequency of publication and type of previous activity. Least-squares regression analysis was used to investigate the relationship between preresidency academic productivity, advanced degrees, stated interest in academics, and other careers and radiology residency publications. RESULTS: There was no statistically significant difference in the number of articles published by those residents who were active and those who were not active before residency (P = .21). Only authorship of papers as an undergraduate was weakly predictive of residency publication. CONCLUSION: These selected measures of academic productivity as an undergraduate and during medical school are not helpful for predicting publication during residency. There was no difference in publication potential between those residents who were academically productive in the past and those who were not. PMID- 11918378 TI - Improving undergraduate education in radiology. PMID- 11918379 TI - On teaching radiology to medical students: a commentary. PMID- 11918380 TI - Organizing the National Institute of Biomedical Imaging and Bioengineering. Part 3. PMID- 11918382 TI - Glucose intolerance by race and ethnicity in the U.S. Virgin Islands. AB - This study describes the prevalence on glucose intolerance by race and ethnicity in the United States Virgin Islands. A population-based sample of 1026 individuals 20 years of age or older was recruited on the island of St. Croix, U.S. Virgin Islands, where 80% of the population classify their race as African American and 20% indicate their ethnicity as Hispanic. American Diabetes Association (ADA) criteria was used to classify glucose tolerance for the entire sample. Persons 40 years of age or older (405) were also administered a 2-h oral glucose tolerance test. Among the major race/ethnic groups, the prevalence of diabetes in patients 20 years of age or older (age-adjusted to the 1995 world population) was 14.1% for non-Hispanic blacks (n = 712), 12.1% for Hispanic blacks (n = 145), 13.5% for Hispanic whites (n = 70) and 1.2% for non-Hispanic whites (n = 37). In each group, the prevalence of diabetes increased with age and appeared higher for men. Among individuals 40 years of age or older a slightly higher prevalence of newly diagnosed diabetes was found when using World Health Organization (WHO) criteria compared to ADA criteria (WHO 10.3%, ADA 7.7% for black non-Hispanic persons and WHO 10.4%, ADA 6.0% for all other groups combined). The prevalence of diabetes for African Americans residing in the U.S. Virgin Islands is similar to rates for the African-American population on the United States mainland and is double that of estimates for blacks on neighboring islands. PMID- 11918381 TI - Acute renal failure mortality in hospitalized African Americans: age and gender considerations. AB - The aging kidney is at risk for both toxic and hemodynamic-induced acute damage, resulting in a high incidence of acute renal failure (ARF) in elderly patients. The effect of age and or gender in ARF mortality in African Americans (AA) was studied in a 3-year, computer assisted retrospective review. In an inner city medical center, 100 patients classified as ARF at discharge or expiration were included in the study. Patients were classified into 3 age categories: <40, 40 64, and >64 years. The incidence of ARF was 35%, 28% and 37%, respectively. Patients >64 years of age were less likely to be dialyzed. Both pre- and postrenal causes of ARF were more common in patients >64 years of age than in younger patients. Hospital length of stay increased progressively with age. Mortality was lower in patients >64 years of age than in younger patients. The incidence of ARF was higher in male than female patients and the incidence of sepsis was higher in female than male patients. Dialytic need was greater in male patients, but mortality was higher in female than male patients. Multivariate logistic regression showed that in the presence of sepsis, oliguria and mechanical ventilatory support, the relative risk of mortality associated with advanced age was 16.5, the relative risk of mortality associated with female gender was 0.2. In summary, hospitalized elderly African-American patients have a high incidence of ARF, and patients less than 40 years of age are equally at risk. Although mortality was higher in female patients, gender and advanced age did not independently contribute to high mortality. Neither age nor gender considerations should supplant sound clinical judgment in the management of and decision making in elderly African-American patients with ARF. PMID- 11918383 TI - Internalized racism, body fat distribution, and abnormal fasting glucose among African-Caribbean women in Dominica, West Indies. AB - The current study examined the relationship of internalized racism to glucose intolerance in a population of Afro-Caribbean women aged 18 to 55. Also of interest was whether this relationship would be differentially influenced by the type of body fat distribution or confounded by the level of hostility. A total of 244 women were selected from a systematic sample of households on the island of Dominica, West Indies. Demographic data together with information on internalized racism were collected by questionnaire. Anthropometric information and fasting blood glucose were also measured. Women with high levels of internalized racism exhibited an increased risk of elevated fasting glucose compared to those with low levels of internalized racism (odds ratio (OR) = 2.4; 95% confidence interval (CI) = 1.1-5.5). There was no difference in mean body mass index (BMI) by level of internalized racism. However those with high internalized racism had a significantly larger waist circumference after adjusting for age, education, hostility, and elevated fasting glucose status. In multivariate analyses controlling for age, education, hostility, and either weight or BMI, internalized racism remained independently associated with elevated fasting glucose. However, once waist circumference was included in the model, the relationship of internalized racism to elevated fasting glucose was not statistically significant. This study demonstrates a significant relationship between internalized racism and abnormal levels of fasting glucose which may be mediated through abdominal fat. The exact nature of the relationship of internalized racism to glucose intolerance may be an important area of future study. PMID- 11918384 TI - Breast cancer racial differences before age 40--implications for screening. AB - BACKGROUND: Most authorities advocate mammogram screening for breast cancer beginning at age 40 based on the age-specific distribution and incidence of breast cancer in the general population. This policy has been bolstered by studies that demonstrate that, for the general population, mammography in the 40 49 age bracket reduces mortality. However, it also has been reported that African American breast cancer patients are diagnosed more often than white patients below the age of 40. Young African-American women are also more likely to have advanced disease at the time of diagnosis with predictably higher mortality. The purpose of this investigation is to explore the question, whether a subset of African-American women, age 30-39, by virtue of increased vulnerability, would benefit from early mammogram screening. STUDY DESIGN: The age-specific distribution (age 30-84) of African-American and white breast cancer patients in five State cancer registries were compared. Prognostic indicators (tumor size and nodal status) in two of the five registries in African-American and white breast cancer cases below the age of 40 were compared. Age-specific incidence in the 30 39 age group and the relative populations of black and white women in the United States were noted in the Surveillance Epidemiology and End Report (SEER) (1994 1998) and The U.S. Census 2000. RESULTS: The differences of age-specific distribution and age-specific incidence of African-American and white breast cancer patients were found to be significant. More than 10% of African-American women with breast cancer were diagnosed before age 40 compared to 5% of white patients. The incidence of breast cancer (SEER Report 1994-1998) in the 30-39-age bracket for African-American and white women was 48.9 and 40.2 at the 95% confidence level, while the proportion of African-American and white women reported by the Census Bureau was not too dissimilar, 15.8% and 14.6% respectively. Prognostic indicators (tumor size and nodal status) support the notion that young African-American women are more likely to have advanced disease at diagnosis. CONCLUSIONS: African-American women in the 30-39 age group have twice the age-specific distribution, have a higher incidence compared to their white counterparts, and exhibit more ominous prognostic signs. This study provides evidence that African-American women in the 30-39 age category represent a high-risk group that may benefit from efforts at earlier detection. Although mammography remains the preferred screening modality, investigators have pointed out difficulties encountered when using mammography in young women, including low sensitivity, high breast density, cost/benefit concerns, and low positive predictive value. Nevertheless, the increasing mortality and persistent racial incidence gap in young African-American women, age 30-39, argue for considering early screening mammography in spite of recognized concerns. PMID- 11918385 TI - Does patient educational level affect office visits to family physicians? AB - Significant disparities in health care based on patient socioeconomic status have been documented. The extent to which physician behavior accounts for these differences is not known. We examined the impact of patient socioeconomic status, measured by years of education, on physician behavior assessed by direct observation of office visits, chart audits, and patient reports among 138 family physicians in 84 practices. Outcomes included time use measured with the Davis Observation Code, delivery of preventive services recommended by the US Preventive Services Task Force, satisfaction assessed with the MOS 9-item Visit Rating Scale, and delivery of attributes of primary care measured by the Components of Primary Care Index. After controlling for patient characteristics among 2538 visits by adult patients who returned questionnaires, a high school education or less was associated with slightly greater proportion of time spent on physical examination and providing nutrition counseling, and less time on patient questions, assessing patient health knowledge, negotiation, and exercise counseling. This indicates that physicians adopt a more directive style with less educated patients. Screening tests were provided at lower rates among less educated patients, but there were no differences in rates of health habit counseling or immunization services delivered and no differences in delivery of patient-reported components of primary care. Less educated patients had similar overall visit satisfaction, but were slightly less likely to have their expectations met. These show that patients' education has relatively small, but potentially important, effects on the outpatient delivery of primary care. PMID- 11918386 TI - A positive antibody screen--an encounter with the Augustine antibody. AB - An antibody screen is performed on the blood of patients who may require blood transfusion. If an antibody is detected, it must be identified to avoid transfusing the patient with blood that contains the corresponding antigen. Antibody screens are also performed as part of a prenatal profile to detect antibodies that may cause hemolytic disease of the newborn. In this article we report the detection of a unique antibody to an antigen of high incidence, the anti-Augustine antibody. We describe problems that may occur when this antibody is encountered, including its identification and obtaining suitable transfusion products for the patient. A brief historical review of the clinical significance of this antibody is included in the article. PMID- 11918387 TI - Exposure to Reach Out and Read and vocabulary outcomes in inner city preschoolers. AB - To examine the association between exposure to Reach Out and Read and vocabulary outcomes in children, a consecutive sample of 200 parent/child pairs was studied at two inner-city health centers. Children at clinic A were exposed to Reach Out and Read, a clinic-based literacy intervention, for 3 years at the time of the study; children at clinic B were unexposed. Main outcome measures were the "Expressive and Receptive One Word Picture Vocabulary Tests" to measure vocabulary in the children and the "Home Literacy Orientation" scale and "READ" subscale of the STIMQ, to measure book-sharing activities. A total of 200 subjects participated, and the mean age of children was 3.8 years. Demographic characteristics were comparable for both clinics at baseline. Exposed children scored higher on receptive vocabulary (81.5 vs. 74.3; p = 0.005). They also scored higher on both the Home Literacy Orientation scale (4.3 vs. 3.3; p = 0.002) and the STIMQ-READ (12.6 vs. 11.0; p = 0.056). There were no differences in expressive vocabulary scores between the two sites (79.5 vs. 77.5; p = 0.26). In conclusion, we found a positive association between exposure to Reach Out and Read and better receptive vocabulary scores. We also found higher scores for Reach Out and Read-exposed children on measures of home reading activities. PMID- 11918389 TI - Genetic weapons. PMID- 11918388 TI - It's time to step up the management of community-acquired pneumonia. AB - Optimal therapy for serious Streptococcal pneumoniae infections with intermediate or high-grade resistance to penicillin is controversial. It should be noted that data regarding the efficacy of penicillins or cephalosporins for penicillin resistant strains are limited. Despite the paucity of clinical trials, most clinicians still agree that penicillins remain the mainstay of therapy for community-acquired pneumonia caused by Streptococcal pneumoniae-susceptible strains. Macrolide antibiotics are effective for treatment of community-acquired pneumonia caused by susceptible strains of Streptococcus pneumoniae. But resistance to all macrolides, based on current National Committee for Clinical Laboratory Standards breakpoints, should be assumed among isolates with erythromycin resistance. The late-generation fluoroquinolones have a role for treatment of community-acquired pneumonia, however, there is also the potential for evolution of antimicrobial resistance. Performance indicators for community acquired pneumonia are being established with implementation of protocols for inpatients with pneumonia. These indicators are being monitored by the Center for Medicare and Medicaid Services (CMS) for medicare patients as part of a national project. The indicators also address documentation of influenza and pneumococcal vaccine status in patients. Several other indicators, such as obtaining blood cultures before antibiotic administration, using antibiotics according to current guidelines, and timely administration of antibiotics, will play critical roles in the management of community-acquired pneumonia. Because of increased incremental costs associated with community-acquired pneumonia, early diagnosis and timely intravenous to oral switch therapy will continue to be emphasized and monitored in those admitted into hospitals, together with the appropriate decision tree based pneumonia specific severity of illness scoring system. PMID- 11918390 TI - Diversity and patient care discussions. PMID- 11918391 TI - British paediatric and American pediatric nurses: how well do we know each other? AB - ISSUES AND PURPOSE: To identify the similarities and differences between pediatric nurses in the United States and Britain, and to compare knowledge of each other's healthcare systems. DESIGN AND METHODS: A descriptive comparative study of 55 U.S. pediatric nurses and 37 U.K. paediatric nurses. Knowledge tests were constructed as parallel forms of items about each healthcare system. "Self knowledge" and "other-knowledge" scores were compared. RESULTS: British nurses' scores were higher for self and other than were U.S. nurses' scores. Individual items suggested that U.K. nurses were more aware of U.S. systems than U.S. nurses were of U.K. systems. PRACTICE IMPLICATIONS: The U.S. and U.K. health systems are equally progressive, yet fundamentally different. To collaborate effectively, we must understand the similarities and differences. PMID- 11918392 TI - Characteristics of school-age children who had colic as infants: mothers' views. AB - ISSUES AND PURPOSE: To describe the behavioral style and personality characteristics of young school-age children who were identified as colicky infants and the parents' perspective of any residual behaviors or effects. DESIGN AND METHODS: A qualitative descriptive design with mothers of 12 children identified with colic as infants and 8 mothers of children who did not have colic. RESULTS: Three themes of children who had colic: independence/self reliance, activity, and emotional lability/intensity. PRACTICE IMPLICATIONS: Children generally were perceived as normal and healthy. Residual feelings, parent-child interaction issues, and perceived or real persistent child characteristics may surface in families who experienced colic, especially during stressful periods. PMID- 11918393 TI - Caring for street youth: a nursing challenge. PMID- 11918394 TI - Evaluating health web sites for research and practice. PMID- 11918395 TI - Assessing children's responses to terrorism. PMID- 11918396 TI - How to help children and adolescents deal with the threat of terrorism. PMID- 11918397 TI - All pediatric nurses are specialists. PMID- 11918398 TI - Rearing the child who is technology dependent: perceptions of parents and home care nurses. AB - ISSUES AND PURPOSE: Most children who are dependent on technology for survival live with their families at home. This study explores the perceptions of parents and home care nurses regarding rearing the technology-dependent child. DESIGN AND METHODS: In this qualitative study, interviews were conducted with 16 parents whose child is technology dependent and 15 registered nurses who provided home care. RESULTS: Rearing the child who is technology dependent is similar to but different from raising other children. Parental communication and negotiation of child-rearing expectations with home care nurses is essential. PRACTICE IMPLICATIONS: Improved collaboration and communication between parents and nurses may reduce parental stress and enhance development for children who are dependent on technology. PMID- 11918399 TI - A method for measuring particle number emissions from vehicles driving on the road. AB - Earlier research has demonstrated that the conditions of dilution of engine exhaust gases profoundly influence the size distribution and total number of particles emitted. Since real world dilution conditions are variable and therefore difficult to simulate, this research has sought to develop and validate a method for measuring particle number emissions from vehicles driving past on a road. This has been achieved successfully using carbon dioxide as a tracer of exhaust gas dilution. By subsequent adjustment of data to a constant dilution factor, it is possible to compare emissions from different vehicles using different technologies and fuels based upon real world emission data. Whilst further optimisation of the technique, especially in terms of matching the instrument response times is desirable, the measurements offer useful insights into emissions from gasoline and diesel vehicles, and the substantial proportion of particles emitted in the 3-7 nanometre size range. PMID- 11918400 TI - Flocculation/aggregation of cohesive sediments in the urban continuum: implications for stormwater management. AB - The urban continuum, as it applies to sediments and associated contaminants, represents the area over/through which sediments are conveyed from a depositional or eroded surface to a treatment system and/or receiving water body. This study has focused on the changing physical characteristics of the sediment, with an emphasis on flocculation/aggregation, as it progresses through the urban continuum. The sediments of the urban continuum are found to change from an unflocculated state on the street, to a flocculated state in the surface runoff to a very large floc form in the sewer system. The high organic content in the sewers contributes to the large floc size. The structure of the flocs and the flow regime of the receiving water will dictate the fate of the sediment following a combined sewer overflow. Probability distributions fitted to the distributions of each sediment type (compartment) confirmed significant differences in the sediment population sizes. Bulk and individual particle settling velocity experiments also revealed substantial differences between compartments. Sewer flocs were found to be of low density, with high porosity, water and organic content and with settling velocities which increase with floc size. PMID- 11918401 TI - Toxicity and color formation during ozonation of mono-substituted aromatic compounds. AB - This research investigates the aqueous toxicity occurring during color formation due to ozonation in a semi-batch reactor. The tested mono-substituted aromatic compounds were aniline, phenol (-NH2' -OH, electron-donating group), nitrobenzene and benzoic acid (-NO2' -COOH, electron-withdrawing group). The results revealed that the aqueous toxicity of the Microtox assay from oxidized samples during the early stage of ozonation increased when the color formation was observed, except in the case of nitrobenzene under high pH conditions. Toxicity typically reached a peak at or near the moment of maximum color intensity. Ozonation of these aromatic compounds showed a large increase in toxicity, and the 50% effective concentration (EC50) expressed as chemical oxygen demand (COD) (mg 1(-1)) decreased to 1.4 (aniline), 1.2 (benzoic acid), 0.8 (phenol) and 0.6 (nitrobenzene) orders-of-magnitude. Moreover, the aromatic compounds with an electron-donating group promoted color formation more than did the electron withdrawing group substituted compounds. PMID- 11918402 TI - Full-scale evaluation of a multi-component additive for efficient control of activated sludge filamentous bulking. AB - A multi-component additive formulated for a more efficient control of activated sludge filamentous bulking was evaluated at a full-scale treatment plant experiencing severe filamentous bulking. It was found that, besides offering an immediate improvement of sludge settling, the multi-component additive was able to eliminate the filamentous bacteria causing the bulking. Hence, contrary to ordinary additives, this novel additive yielded immediate as well as long-term improvements in sludge sedimentation upon a few additions. Preliminary lab-scale toxicity tests showed that the treatment of the sludge by the additive should not impart any toxicity to the resulting effluent. PMID- 11918403 TI - The monitoring of a two step aerobic-anoxic process with separate biomass to enhance performance in the treatment of liquid industrial wastes. AB - The paper presents the results of a one-year study of the performance of a full scale plant for the treatment of industrial liquid wastes adopting the alternate cycle process. The carbon and nitrogen removal performances were discussed according to the experimental measurements of maximum nitrification and denitrification rates. It was demonstrated that the nitrification process was the limiting step: it worked with a rate in the range 0.002 - 0.02 KgNH4-N kg(-1)VSS d(-) at 20 degrees C. This was because of inhibition phenomena due to the presence of both complex organic compounds and heavy metals which were not removed by the chemical-physical pre-treatment step. The denitrification process was characterized by a maximum rate ranging from 0.015 to 0.056 Kg NO3-N kg( 1)VSS d(-1) at 20 degrees C, according to the available amount of readily biodegradable COD in the treated wastes. The reliability of the aerobic-anoxic process was determined on the basis of the percentage of successful cycles compared with the performed ones. It was shown that the actual cycles ranged from 50 to 100% of the expected ones, while effective cycles were up to 84% in the first step and up to 60% in the second one. These were related to the carbon to nitrogen ratio. Even if at times the nitrogen and carbon removal yields were not satisfactory, the two step aerobic-anoxic process operated in the alternate cycle mode seems a successful solution for the treatment of liquid industrial wastes. PMID- 11918404 TI - Cr(VI) removal in acidic aqueous solution using iron-bearing industrial solid wastes and their stabilisation with cement. AB - In this study, iron-bearing industrial solid wastes iron filings, ETP sludge of steel and red mud of aluminium industries; were used for Cr(VI) removal at pH 3. A complete removal of Cr(VI) was found for initial 10 mg 1(-1) of 100 ml solutions in the presence of 2.5 g iron filings, 8 g ETP sludge and 10 g red mud for up to one hour of shaking at room temperature. After Cr(VI) removal, inclusion of chromium on the reacted iron filing surface was demonstrated by EDAX analysis. Leachability of chromium and iron from the reacted wastes was determined by using Toxicity Characteristics Leaching Procedure (TCLP). This test showed a very low level of leachability of chromium as Cr(III) and iron from the reacted wastes. To minimise their leachability further, Cr(VI)-reacted solid wastes were stabilised with Portland cement in their 3:1 ratio. Leachability tests of stabilised wastes by TCLP indicated a considerable decrease in leachability of chromium and iron compared with the that of reacted wastes alone. To explore the possibility of utilisation in building materials, bricks of cement mixed Cr(VI)-reacted wastes were made and their comprehensive strength, durability and leachability under immersion conditions were measured. PMID- 11918405 TI - Development of fixed bed adsorber for abatement of endosulfan in aqueous media. AB - In this paper an attempt was made to develop and evaluate the performance of a fixed bed adsorber (FBR) column to remove endosulfan, an organochlorine pesticide from the water environment using wood charcoal as an adsorbent. The FBR column design parameters were evaluated using the column breakthrough data at different bed depths. The Bohert and Adams model was used with bed depth-service time (BDST) approach for the design of the adsorber. Similarity in service time values estimated from BDST approach and experimental curves revealed that BDST approach could be adapted for the design of this system. Desorption of the exhausted column with 10% methanol solution took lhr with maximum desorption occurring at 0.5 hrs with a maximum endosulfan concentration of 47 mg 1(-1). A laboratory column study for 3 successive cycles of operation (adsorption-desorption) revealed that the column could treat 47.27 bed volumes of endosulfan contaminated water up to breakthrough and 158.45 bed volumes up to exhaustion. It showed a promising performance with more than 89% of its original efficiency even after the 3rd cycle of operation. It dearly demonstrates adaptability of the present study in developing adsorber columns for rural areas. PMID- 11918407 TI - Application of nano-diffraction to local atomic distribution function analysis of amorphous materials. AB - A method is proposed for the local atomic distribution function analysis of amorphous materials. This method is based on local halo-electron diffraction intensity analysis with nano-sized electron probes as small as 25 to approximately 3 nm, taking advantage of the intensity recording with imaging plate. Nanodiffraction and selected area electron diffraction (SAED) patterns from an amorphous SiNx (x approximately 4/3) thin film were taken using a conventional transmission electron microscope operated at 200 kV and recorded on imaging plates. An intensity correction to omit inelastic intensity was made using electron energy-loss spectroscopy. When a beam-convergence angle is larger than 1 x 10(-3) rad, the Wiener-filter deconvolution method becomes helpful in producing atomic pair distribution functions (PDFs) from the nano-diffraction intensity profiles that are more similar to the PDF from the SAED intensity. This technique was applied to the analysis of local amorphous structures of SiO2 layers formed by an oxygen-ion implantation into single crystal SiC. PMID- 11918406 TI - Structure of [110] tilt grain boundaries in zirconia bicrystals. AB - Cubic stabilized zirconia bicrystals with [110] symmetric tilt grain boundaries were fabricated by diffusion bonding of two single crystals with the composition of ZrO2-9.6mol%Y2O3. The structures of symmetric tilt small angle grain boundary and two types of symmetric tilt sigma3 grain boundaries with different grain boundary planes were observed by transmission electron microscopy (TEM). High resolution transmission electron microscopy (HREM) observations clarified that the [110] small angle tilt grain boundary consists of periodic array of b = a/2[110] type edge dislocations. This result is consistent with Frank's dislocation model for small angle grain boundary. HREM observation also revealed that the 70.5 degrees sigma3 grain boundary shows atomically coherent grain boundary structure with the boundary plane of [111], while the 109.5 degrees sigma3 grain boundary accompanies grain boundary facets taking [111]/[115] asymmetric grain boundary plane. Because of the very low surface energy of [111] plane and/or high lattice matching of [111] and [115] type planes, the grain boundary faceting may be preferred in spite of increasing grain boundary area to about 6%. TEM-energy-dispersive X-ray spectroscopy (EDS) analyses were performed on both sigma3 grain boundaries, and the segregation of yttrium ions to the boundaries was detected in both cases. The amount of segregation is about the same in both sigma3 boundaries. It can be concluded that the segregation of yttrium ions to sigma3 grain boundary exists in cubic zirconia. PMID- 11918408 TI - Identification of various phases in HRTEM images of MgO-PSZ. AB - Magnesia partially stabilized zirconia (MgO-PSZ) is one of the most commonly used engineering ceramics based on zirconia. A detailed discussion about how to identify the various phases in high-resolution transmission electron microscopy images of this material is presented. It shows that in some cases, standard procedures of image simulation are inadequate to interpret these images. By including the effect of astigmatism in both experimental and simulated images, together with the digital Fourier transforms of the images, orthorhombic ZrO2 in [001] orientation was identified. The delta-phase, which has a marked effect on the thermomechanical properties of MgO-PSZ, can most easily be identified by high resolution imaging in the [130]c zone that coincides with a low-index zone axis of the delta-phase. PMID- 11918409 TI - Understanding magnetic structures in permanent magnets via in situ Lorentz microscopy, interferometric and non-interferometric phase-reconstructions. AB - We present our observations of field- and orientation-dependence of magnetic domains and their reversal nucleation at grain boundaries in polycrystalline Nd2Fe14B, using Fresnel- and Foucault-Lorentz microscopy. The local magnetization associated with the domain and domain-wall in Nd2Fe14B and in precipitated Fe particles was mapped using an interferometric holography as well as a novel non interferometric method based on the 'transport of intensity' equation. PMID- 11918410 TI - HRTEM study of new series of oxycarbonitrate superconductors (Cu,C,N)Sr2Ca(n - 1)Cu(n)O(y) (n = 1-6). AB - Arrangements of Cu and anion groups (CO3 and NO3) in the charge-reservoir (CR) blocks of a series of new oxycarbonitrate superconductors (Cu,C,N)Sr2Ca(n - 1)Cu(n)O(y) (n = 1-6) were examined by means of electron diffraction and high resolution transmission electron microscopy (HRTEM). The first three members with n = 1-3 [Tc = 33 K (n = 1), 91 K (n = 2), 90 K (n = 3)] show the 4a0-type superstructures with periodic arrangements, [-Cu-X-X-X-Cu-X-X-X-Cu-] (X = CO3, NO3), in the CR blocks. The third member (n = 3) partly contains the 2a0-type of superstructure with [-Cu-X-Cu-X-Cu] in the CR blocks. The fourth member with n = 4 (Tc = 113 K) contains only the 2a0-type of superstructure. The higher members, with n = 5 (Tc = 65 K) and n = 6 (Tc = 52 K), show no evidence of ordering in the CR blocks, suggesting random arrangements of Cu and anion groups. PMID- 11918411 TI - A novel shadow-imaging technique to measure charge distribution and lattice displacement. AB - We developed a novel shadow-imaging diffraction technique, using both non coherent and coherent sources, based on parallel recording of diffraction intensity of many reflections to measure charge distribution and lattice displacement in crystals and in defects. Applying the method to Bi2Sr2CaCu2O8 superconductors demonstrated its unprecedented sensitivity and accuracy. PMID- 11918412 TI - TEM and HAADF-STEM study of a Au catalyst supported on a TiO2 nano-rod. AB - Gold nanoparticles were deposited on TiO2 nano-rods by a deposition precipitation method. This Au/TiO2 nano-rods catalyst is active for CO oxidation even at temperatures less than 273K and is as active as the catalyst of Au supported on conventional TiO2 particles. Transmission electron microscopy (TEM) showed that the TiO2 nano-rods were composed of a single anatase crystal and had a pillar shape with an axis along the <111> direction surrounded by flat four [101] planes. High angle annular dark field scanning TEM (HAADF-STEM) also revealed that a Au platelet with a thickness of 0.5-1 nm was formed at the interface between TiO2 nano-rods. The Au platelet was not observed when conventional round particles of TiO2 were used as a support. PMID- 11918413 TI - Examination of electrostatic potential distribution across an implanted p-n junction by electron holography. AB - In the manufacture of semiconductor microelectronic devices, a p-n junction is formed usually by implanting a high concentration of impurity into a less heavily doped region and then heat annealing. A Si/Si p-n junction test sample has been made following the above practical process and thinned for electron holographic observation by using argon ion-milling. From the reconstructed phase image, the phase shift induced by potential drop across p-n junction can be seen clearly. To characterize quantitatively this potential drop, the mean inner potential V0 of silicon was measured precisely by electron holographic method. By measuring 25 different crystalline silicon spheres with diameter ranging from 40 to 170 nm, an average result of V0 = 12.16 +/- 0.83 V was obtained. By using this V0 value, a quantitative measurement yields the potential drop approximately 0.70 V, which is reasonably consistent with expected Si/Si junction parameter. The thickness of electric dead layer in depletion region produced from this measuring is approximately 20 nm on each sample surface. PMID- 11918414 TI - Electron transport behaviour in Nb-doped SrTiO3 bicrystals. AB - In order to elucidate the relationship between electron transport behaviour and defect chemistry, grain boundary structure and current-voltage characteristics across the boundary were investigated for Nb-doped SrTiO3 bicrystals. Two kinds of boundaries, i.e. small angle and random boundaries, were prepared for 0.2at% and 1.0at% Nb-doped SrTiO3. The bicrystals were prepared by joining two single crystals at 1400 degrees C for 10 h under a pressure of 0.4 MPa in air. High resolution transmission electron microscopy (HRTEM) study revealed that all of the joined boundaries are free from any secondary phases or amorphous films. On the other hand, it was found that non-linearity in current-voltage dependence becomes remarkable by reduction of cooling rate after joining in small angle boundaries of 0.2at% Nb-doped SrTiO3 bicrystal. In addition, the random boundary of 1.0at% Nb-doped SrTiO3 bicrystal exhibits clear alpha = 2 I-V relation, which appears across a contact of semiconductor-insulator-semiconductor (n-i-n). From the results of HRTEM study and I-V behaviours, it could be concluded that the electron transport mechanism is controlled mainly by defect chemistry and not by the grain boundary structure. PMID- 11918415 TI - Analysis of contacts and V-defects in GaN device structures by transmission electron microscopy. AB - This paper describes the microstructure of ohmic contacts to an AlGaN/GaN heterostructure, of interest for high power transistors, and an analysis of V defects in an InGaN/GaN multi-quantum well (MQW) light-emitting structure. A combination of different transmission electron microscopy (TEM) techniques has been employed, as they provide complementary information. These include bright field and dark field TEM, high-resolution electron microscopy, X-ray mapping, energy filtered TEM and high angle annular dark field. A full determination of the phase distribution in the ohmic contacts was achieved. The onset of low contact resistance was found to correspond with the formation of an interfacial layer containing both TiN and AlN, and of an intermetallic layer containing Al, Ti and Au in contact with it. The MQW structures were capped with a p-type GaN layer, and TEM and ADF studies of the samples show a number of V-defects 100-200 nm apart along the MQW. Each V-defect incorporates a pure edge (b = 1/3 <11-20>) dislocation, which runs through its apex up to the free surface. The defects contain GaN with no InGaN layers, suggesting the V-pits have been filled in by the capping layer. PMID- 11918416 TI - Four-dimensional dielectric property image obtained from electron spectroscopic imaging series. AB - We have demonstrated a new quantitative method to characterize two-dimensional distributions of energy-dependent dielectric function of materials from low loss electron spectroscopic image (ESI) series. Two problems associated with extracted image-spectrum from the low-loss image series, under-sampling and loss of energy resolution, were overcome by using fast Fourier transformation (FFT) interpolation and maximum entropy deconvolution method. In this study, Black Diamond/Si3N4/SiO2/Si-substrate dielectric layer designed for copper metallization was used as the sample. We show that the reconstructed (FFT interpolated and maximum entropy deconvoluted) image-spectrum obtained from ESI series images can be quantified with the same accuracy as conventional electron energy-loss spectroscopy spectra. Since the analysis of the dielectric function is sensitive to the local thickness of the specimen using Kramers-Kronig analysis, we also developed a new method to quantitatively determine the dielectric constant for low-k materials. We have determined the thickness of the Black Diamond using the extrapolated thickness method from the materials of known dielectric constants. Using Kramers-Kronig formula, the dielectric function map can be deduced from two-dimensional reconstructed single scattering spectra with providing the information of thickness. We proposed a four-dimensional data presentation for revealing the uniformity of the energy dependent property. The accuracy of our methods depends on the thickness determination and on the quality of the reconstructed spectra from the image series. PMID- 11918417 TI - Advances in energy-filtering transmission electron microscopy. AB - First experiments using a new energy-filtering microscope (Sub-eV-Sub-A microscope, or SESAM) are shown. They make use of the high transmissivity of the 90 degree filter. This allows the mapping of chemical bonding of large specimen areas, even if narrow energy selecting slits are used. Because large scattering angles are accepted by this filter, energy-filtered diffraction patterns extending to 150 mrad can be recorded by a single exposure. This can be used to determine the reduced density function of amorphous materials and reduces the exposure time of the investigated area by three orders of magnitude as compared with previous approaches. PMID- 11918418 TI - Artificial bright spots in atomic-resolution high-angle annular dark field STEM images. AB - Artificial bright spots, which appear in some atomic resolution high-angle annular dark field scanning transmission electron microscope (HAADF STEM) images, have been accounted for by simulations based on Bloch wave description. This is illustrated with Si and SrTiO3 images. The simulation reveals that bright spots on no-atomic columns in [011]-orientated Si images are produced by thermal diffuse scattering from Si atoms on their surrounding atomic columns, which are under the subsidiary peaks in the incident convergent electron probe. Similarly, bright spots on oxygen columns in [001]-orientated SrTiO3 images are ascribed to Sr and Ti atoms in their surrounding atomic columns rather than O atoms in the O columns. The probe function, therefore, provides a simple explanation for the appearance of these artificial spots. PMID- 11918419 TI - Spatially-resolved EELS analysis of multilayer using EFTEM and STEM. AB - Spatially-resolved electron energy-loss spectroscopy (EELS) is applied to the multilayer (SiO2/Si3N4/SiOxNy/Si), a common semiconductor device structure. To observe depth profile at sub-nanometre spatial resolution, scanning transmission electron microscopy (STEM) and energy-filtering transmission electron microscopy (EFTEM)-based techniques are compared in practical application. STEM-based EELS is useful in simultaneously analysing plural elements. EFTEM-based EELS is suitable for analysing the chemical shift and core loss intensity of single element. The practical spatial resolution is of the same order for each technique, both involving beam damage, which can be reduced by decreasing current density in EFTEM and avoiding beam overlapping in STEM. PMID- 11918420 TI - Extension of HRTEM resolution by semi-blind deconvolution method and Gerchberg Saxton algorithm: application to grain boundary and interface. AB - A generalized maximum entropy method coupled with Gerchberg-Saxton algorithm has been developed to extend the resolution from high-resolution TEM image(s) for weak objects. The Gerchberg-Saxton algorithm restores spatial resolution by operating real space and reciprocal space projections cyclically. In our methodology, a generalized maximum entropy method (Kullback-Leibler cross entropy) dealing with weak objects is used as a real space (P1) projection. After P1 projection, not only are the phases within the input spatial frequencies improved, but also the phases in the next higher frequencies are extrapolated. An example of semi-blind deconvolution (P1 project only) to improve the resolution in SiC twin boundary is shown. The nature of the bonding in this twin boundary is Si-C but it was rotated 180 degrees along the boundary normal. The optimum solution from P1 projection can be further improved by a P2 projection. The square roots of diffraction intensities from a diffraction pattern are then substituted to complete a cycle operation of the Gerchberg-Saxton algorithm. Application examples of Gerchberg-Saxton algorithm to solve the atomic structure of defects (2 x 1 interfacial reconstruction and dislocation) in NiSi2/Si interfaces will be shown also. PMID- 11918421 TI - High-resolution transmission electron microscopy observation of the cross sectional structure of reconstructed silicon (5,5,12) surface. AB - The cross-sectional reconstructed structure of Si (5,5,12) surface was, for the first time, observed using ultrahigh vacuum high-resolution transmission electron microscopy (UHV-HRTEM) profile view method. In the high-index region, two units of the (337) surface combine with one units of the (225) surface to complete one unit cell of (5,5,12) surface. The (337) unit at one side of the (225) unit is distinctly different from that at another side of (225) unit. The observed HRTEM images do not agree with the previous structural models of the (5,5,12) surface. We propose a new structure model of this surface using a TEM image simulation. Our model is agreement with the previously reported scanning tunnelling microscope images. PMID- 11918422 TI - Reactive epitaxy of Co nanoparticles on (111)Si. AB - The formation of epitaxial CoSi2 islands of nanoscopic dimensions is reported using the technique of reactive cluster deposition. Co clusters in the size range 5-50 nm were synthesized by sputtering of a high purity Co target inside a ultra high vacuum (UHV) sputtering chamber, using the technique of inert gas condensation. The clusters were then deposited on the reconstructed Si (111) surface. Upon annealing, the particles reacted with the Si substrate to form epitaxial CoSi2. Our observations were made using a JEOL 200CX transmission electron microscope modified for in situ sputtering and UHV conditions. PMID- 11918423 TI - The consequences of imbibing alcohol in the absence of adequate nutrition: the salt and water hypothesis. AB - Alcohol can have an effect on almost every cell in the human body and it is becoming increasingly clear that when alcohol is consumed the prior nutritional status of the individual may be an important factor for long-term health. The salt and water hypothesis integrates the biochemical findings from the current alcohol literature and proposes a mechanism by which alcohol consumption prior to food intake may cause a transient alteration in the functioning of the hypothalamic-pituitary-adrenal axis due to an alcohol-induced impairment in electrolyte regulation. PMID- 11918424 TI - GABA-transaminase, the liver and infantile autism. AB - Gamma-aminobutyric acid (GABA) is a major inhibitory neurotransmitter of the mammalian brain and the enzyme responsible for catabolism (breakdown in the liver during regulation) is GABA-Transaminase (GABA-T). Recently, Cohen has shown that extremely high GABA levels in the urine and blood were observed for an autistic child. The finding that elevated levels of GABA in the urine and blood are present for an autistic child could explain why autistic features (such as self stimulatory behavior and language delays, etc.) are found. Increasing the GABA-T enzyme activity for this autistic patient could result in less plasma GABA (after liver regulation) entering into the bloodsteam and brain and it is postulated that this could result in a reduction of the autistic features (such as self stimulatory behavior and language delays, etc.) due to abnormal development of the axon(s) in the corpus callosum. PMID- 11918425 TI - Fresh frozen bone in femoral impaction grafting: can developments in bone regeneration improve on this? AB - Major advances in the understanding of bone regeneration have led to revolutionary applications of bone conduction and induction. Revision hip surgery is on the increase with an active, ageing population demanding increased quality of life. The technically demanding instrumentation and materials necessary for revision prosthetic hip replacements have been rapidly incorporated into clinical practice. Unfortunately, advantage has not been taken of the basic and fundamental principles of bone induction and regeneration, available for many years now. In impaction grafting for revision hip replacements where fresh frozen bone is currently used as a standard, we argue and demonstrate from the literature why this technique is biologically inferior to the results which potentially can be gained by exploiting osteoinductive grafts such as demineralised bone matrix, hydroxyapetite, and inductive factors such as bone morphogenetic proteins. PMID- 11918427 TI - HIV infections and AIDS development: the role of adjuvant activation. AB - The human immunodeficiency virus (HIV) has probably coexisted with humans for nearly a century. However, towards the end of the 1970s, it appeared as the etiologic agent of the acquired immunodeficiency syndrome (AIDS). In this paper, we propose a hypothesis for HIV infection and AIDS development, considering the role of secondary adjuvant infections. This hypothesis takes into account the natural history of the development of the AIDS epidemic from the standpoint of an emerging infection due to the increased risk caused by anthropogenic activities, and it also considers the progression of the infection to the syndrome, in terms of cellular population dynamics. PMID- 11918426 TI - Manic depressive psychosis and schizophrenia are neurological disorders at the extremes of CNS maturation and nutritional disorders associated with a deficit in marine fat. AB - The maturational theory of brain development comprises manic depressive psychosis and schizophrenia. It holds that the disorders are part of human diversity in growth and maturation, which explains their ubiquity, shared susceptibility genes and multifactorial inheritance. Rate of maturation and age at puberty are the genotype; the disorders are localized at the extremes with normality in between. This is based on the association between onset of puberty and the final regressive event, with pruning of 40% of excitatory synapses leaving the inhibitory ones fairly unchanged. This makes excitability, a fundamental property of nervous tissue, a distinguishing factor: the earlier puberty, the greater excitability--the later puberty, the greater deficit. Biological treatment supports deviation from the norm: neuroleptics are convulsant; antidepressives are anti-epiletogenic. There is an association between onset of puberty and body build: early maturers are pyknic broad-built, late ones linearly leptosomic. This discrepancy is similar to that in the two disorders, supporting the theory that body-build is the phenotype. Standard of living is the environmental factor, which affects pubertal age and shifts the panorama of mental illness accordingly. Unnatural death has increased with antipsychotics. Other treatment is needed. PUFA deficit has been observed in RBC in both disorders and striking improvements with addition of minor amounts of PUFA. This supports that dietary deficit might cause psychotic development and that prevention is possible. Other neurological disorders also profit from PUFA, underlining a general deficit in the diet. PMID- 11918429 TI - Consideration of mycophenolate mofetil for idiopathic pulmonary fibrosis. AB - Idiopathic pulmonary fibrosis (IPF) is an inflammatory disorder of the lungs of unknown etiology, with no effective treatment. Besides the recent finding of utility of mycophenolate mofetil (MMF) in a case of refractory interstitial lung disease associated with ulcerative colitis, I suggest that there are at least three other reasons to consider MMF for IPF. Previously, MMF has been found to be effective as salvage therapy in a number of diseases. MMF might work for IPF not only by white cell suppression, but also in vivo against proliferation of primary human pulmonary fibroblasts. There is one group of patients for whom, logically, MMF should be most strongly considered--those with a high likelihood of receiving a lung transplant. As MMF is often part of the post-transplant immunosuppressive regimen in these patients, logic would seem to dictate MMF should be considered for use before subjecting the patient to major surgery. PMID- 11918428 TI - Stem cell strategies for neuroreplacement therapy in Alzheimer's disease. AB - The existence of neural stem cells (NSCs) in the adult human brain provides impetus for investigating possible neuroreplacement therapies for neurodegenerative disease. Due to recent advances in techniques affording isolation and maintenance of NSCs using non-serum culture media, these cells have become exciting candidates for therapeutic strategies. We are able to expand NSCs by mitogenic growth factors in vitro and in defined conditions, NSCs differentiate into each of the diverse brain cell types: neurons, astrocytes and oligodendrocytes. This article addresses the involvement of amyloid-beta precursor protein and the presenilins in NSCs' biology and possible application of NSCs for therapeutic approaches in Alzheimer's disease. Ongoing studies in our laboratory, and recent findings by others using human neural progenitors, serve as the conceptual frame for this article. PMID- 11918430 TI - Consideration of vaccination against the FSGS factor. AB - Focal segmental glomerulosclerosis (FSGS) is a common cause of nephrotic disease in adults, and it is thought to be increasing in incidence. FSGS is often difficult to treat and even has a high propensity to relapse after transplant. Indeed, this suggests that there might be some circulating factor responsible for proteinuria ('FSGS factor'). Here I suggest that if and when the FSGS factor is found, vaccination against this factor might be a useful treatment modality for FSGS. PMID- 11918431 TI - Tension headaches and muscle tension: is there a role for magnesium? AB - Although many theories and hypotheses have been offered for the etiology of tension-type headache (TH), no one previous hypothesis seems to adequately explain TH. This may, in large measure, account for why it is often difficult to effectively treat TH. Herein, we review current and old hypotheses of TH and offer a new hypothesis which is consistent with what is known about TH. We show that magnesium (Mg) metabolism may be pivotal in both the etiology and treatment of TH. Measurement of serum ionized Mg2+ (IMg2+) levels and brain intracellular free Mg2+ ([Mg2+]i) appear to offer excellent methods for establishing the validity of our hypothesis. Since approximately 70% of patients who have a TH exhibit muscular tightness and tenderness, it is distinctly possible that problems in Mg metabolism and dietary intake are the links to concomitant muscle tension and TH. The significance of release of pain mediators, muscle cramps, muscle strains (and damage) and muscle tension to TH, and its relationship to Mg metabolism, are reviewed. These are all associated with a Mg-deficient state. It seems clear from the available data that TH's are more associated with muscle tension or scalp tension than any other headache type. From the data available, Mg supplementation appears to be of great benefit in many of these situations. We believe there is a great need for clinicians to examine Mg2+ metabolism, bioavailable Mg2+ in muscle tissues and blood, and the effectiveness of Mg salts (in a double-blinded, placebo-controlled manner) in subjects with TH and muscle tension. PMID- 11918432 TI - Anterior insular cortex: linking intestinal pathology and brain function in autism-spectrum subgroups. AB - Autism includes deficits in communications skills and is associated with intestinal pathology. Numerous parents and some physicians report that an autistic child's attention and language improve in response to treatments which eliminate certain dietary antigens and/or which improve intestinal health. For at least some autism-spectrum children, the link between intestinal pathology, attention, and language may derive from shared neuroanatomic pathways within the anterior insular cortex (aIC); from a neurotrophic virus such as herpes simplex (HSV) migrating within afferents to the insular cortex; and/or from synaptic exhaustion in the aIC as induced by chronically inappropriate neuronal activity in the enteric nervous system and/or its vagal efferents. PMID- 11918433 TI - Psychopathology and the binding problem. AB - Normal cerebral activity is based on the rapid and flexible alternation of an enormous quantity of spatial-temporal patterns of neuronal firing. There is still much to learn about the mechanism that keeps neurons on functionally closed assemblies firing in synchronization. The explanation of this mechanism is the so called 'binding problem', one of the cornerstones of the generation of perceptions, cognitions, emotions and behaviours by the brain, and therefore of great interest to psychiatry. We explore the possibility that a 'binding dysfunction' could be a relevant aspect of the pathophysiology of mental disorders, and apply it to schizophrenia, in an attempt to integrate some neurobiological findings in this disorder, including a few preliminary data obtained with transcranial magnetic stimulation (TMS). PMID- 11918434 TI - The ketogenic diet may have mood-stabilizing properties. AB - The ketogenic diet, originally introduced in the 1920s, has been undergoing a recent resurgence as an adjunctive treatment for refractory epilepsy, particularly in children. In this difficult-to-treat population, the diet exhibits remarkable efficacy with two-thirds showing significant reduction in seizure frequency and one-third becoming nearly seizure-free. There are several reasons to suspect that the ketogenic diet may also have utility as a mood stabilizer in bipolar illness. These include the observation that several anticonvulsant interventions may improve outcome in mood disorders. Furthermore, beneficial changes in brain-energy profile are noted in subjects on the ketogenic diet. This is important since global cerebral hypometabolism is a characteristic of the brains of depressed or manic individuals. Finally, the extracellular changes that occur in ketosis would be expected to decrease intracellular sodium concentrations, a common property of all effective mood stabilizers. Trials of the ketogenic diet in relapse prevention of bipolar mood episodes are warranted. PMID- 11918435 TI - Pyrogenic reactions following gentamicin therapy: an alternative explanation. AB - A hypothesis is proposed, arguing that gentamicin administration in a single daily dose results in higher peak tissue concentration, marked bacteriolysis with endotoxin (ET) release and consequent ET-mediated febrile host responses, which resemble Jarisch-Herxheimer reactions. PMID- 11918436 TI - The role of the host-tumor interface and cell hybridization in invasive cancer. AB - Available data indicate that growth of invasive tumors is enhanced by homeostatic mechanisms of the host involved in normal tissue regeneration and repair. To achieve this, malignant cells may (i) induce degeneration of normal cells at the host-tumor interface, (ii) hybridize in situ with activated host stem cells, required for replacement of lost mature tissue cells, (iii) the resulting malignant/normal cell hybrids may exhibit an antigenic similarity to normal cells, (iv) thereby preventing recognition by the immune system, (v) and exploiting normal mechanisms of tissue regeneration by the host. In addition, primary cancers with allotypic determinants may utilize other homeostatic mechanisms evolved in mammals to promote fetal allograft survival. They may have a potential to grow in another (secondary) host. Novel approaches to cancer prevention and control may depend on a better understanding of the mechanisms by which normal cellular growth are controlled, and hybridization prevented. PMID- 11918437 TI - Xerosis in hypothyroidism: a potential role for the use of topical thyroid hormone in euthyroid patients. AB - Xerosis is an extraordinarily common problem in dermatology. Despite the knowledge of well recognized aggravating factors, its etiology is an enigma, and the management of the condition is often suboptimal. Dry skin may be a manifestation of hypothyroidism. The nature of this association is reviewed, culminating in the speculation that topical thyroid hormone may represent a useful modality in euthyroid patients with xerosis or other disorders of keratinization. PMID- 11918438 TI - Ammonia functions as a regulatory molecule to mediate adjustments in glomerular filtration rate in response to changes in metabolic rate. AB - In mammals, varying in size from mouse to elephant there is a close linkage between metabolic rate (MR) and glomerular filtration rate (GFR). This linkage has evolved because of the necessity to excrete nitrogenous wastes generated predominately by amino acid catabolism. A similar linkage is observed in birds, reptiles and fish, although in the latter vertebrate group gill excretion rate and GFR need to be considered as an interdependent co-ordinated system. A change in the rate of amino acid catabolism will result in a change in the rate of production of nitrogenous wastes requiring excretion and hence GFR (or GFR plus gill excretory rate in fish) must be appropriately re-set. It is hypothesized that ammonia functions as a regulatory molecule to mediate adjustments in excretory rate (GFR or GFR plus gill excretory rate) in response to such changes in amino acid catabolism. PMID- 11918439 TI - Physiological rationale of aggressive behavior: a brain blood perfusion hypothesis. AB - The physiological rationale of aggressive behavior is discussed. The potential importance of homeostatic reaction in brain blood perfusion is described. The author speculates that pathological aggressive behavior arises from urgent biological needs. Attacks of anger show increased regional cerebral blood flow in the temporal cortex or other paralimbic areas, which show hypoperfusion in inter attack states. This hypoperfusion may also be related to psychological stress induced cerebral vasoconstriction. Furious physical motion, accompanying the attack, would augment regional cerebral blood flow and maintain it longer. A brain blood perfusion hypothesis as the etiological role of aggressive behavior is presented. PMID- 11918440 TI - Centenarian scientists: an unusual cluster newly formed in the 20th century. AB - From biographical data sources on ranking scientists, I was able to identify 35 centenarians. Among these, only one (Michel Chevereul from France) lived before the 20th century. Since the remaining 34 individuals became centenarians only from 1965, I propose that centenarian scientists are an unusual cluster, first formed in the 20th century. Among these, all except one (Alice Hamilton) were men. Six centenarian scientists, including Hamilton, had received professional medical training. The nationality ranks of the 34 centenarian scientists identified in the 20th century show 26 Americans, 6 British, one German and one French. Four of the 26 Americans were immigrants from Europe. At least three centenarians, namely Michael Heidelberger, Nathaniel Kleitman and Victor Hamburger, belong to the 'Nobel class' category, being pioneers in the disciplines of immunochemistry, sleep physiology and neuroembryology respectively. PMID- 11918441 TI - On new hypotheses about autosomal dominant polycystic kidney disease type 1. AB - The purpose of this paper is to suggest a partial explanation of the aetiology of autosomal dominant polycystic kidney disease type 1, one of the most common genetic diseases in humans. To this aim we put forward a number of interconnected ideas, based on a number of experimental evidences and plausibility arguments. We stress the major role played by the instability of some genomic tandem repeats, together with the DNA structures known as quadruplexes, the pseudogenes and the gene conversion. The model we propose can be considered a multi-hit generalization of the well-known two-hit model, a generalization that could well have a validity also outside the specific context. We finally provide an indication of the likely guilty DNA segment for the above disease, and we propose a possible simple experimental line of action aimed to confirm or disproof our suggestion. PMID- 11918442 TI - Inspection time: a neuropsychophysiological test for measuring the functional integrity of the cholinergic system. AB - Inspection time (IT) is a measure of the speed of early information processing, demonstrating reliable correlation with psychometric intelligence. We now provide evidence that it may serve as a marker for the integrity of the cholinergic system. Cholinergic agonists improve and antagonists impair IT, while modulators of the monoamine systems are without effect. Furthermore impairments in IT correlate with disease severity in Alzheimer's disease, a disorder of compromised cholinergic system function. Taken together, available data suggest that IT may be a useful physiological marker of functional status of the cholinergic system. PMID- 11918444 TI - Are superoxide and/or hydrogen peroxide responsible for some of the beneficial effects of hyperbaric oxygen therapy? AB - The basic mechanisms behind the pharmacologic effects of hyperbaric oxygen therapy are not clear. Reactive oxygen metabolites are generally associated with the adverse reactions to hyperbaric oxygen exposure but they are also believed to be involved in the antibacterial effects of this therapy. The possibility that reactive oxygen metabolites are responsible for some of the other reported beneficial effects of hyperbaric oxygen therapy has not been investigated. This hypothesis paper briefly reviews the literature suggesting that the pharmacologic actions underlying some of the beneficial effects of hyperbaric oxygen therapy may be caused by superoxide and/or hydrogen peroxide. Elucidation of the pharmacologic mechanisms is fundamental in order to fully exploit the therapeutic potential of hyperbaric oxygen and we incite experimental research to be done within this area. PMID- 11918443 TI - The ratio of 2nd to 4th digit length: a proxy for testosterone, and susceptibility to HIV and AIDS? AB - The incidence of HIV and AIDS is high in sub-Saharan Africa and in male homosexuals. It is suggested that testosterone-mediated immune dysfunction may account for this pattern. The ratio of the length of the 2nd and 4th digit (2D:4D) is negatively related to prenatal and adult testosterone concentrations. There is evidence that black South Africans have lower 2D:4D ratios than most other populations and male homosexuals have lower 2D:4D ratios than male heterosexuals. Men with low 2D:4D ratios may also be more sexually active and/or more fertile than men with high ratio. We suggest that men and women with low 2D:4D are susceptible to HIV infection and AIDS and babies with low 2D:4D ratio susceptible to vertical transmission. Infection rates may therefore be reduced by education and condom supply directed towards low 2D:4D adults. PMID- 11918445 TI - A hypothesis of the stomatocytosis in individuals with the phenotype Rh(null). AB - A hypothesis of the stomatocytosis in individuals with the Rh(null) phenotype is proposed on the assumptions that the RhAG polypeptide of the Rh antigenic complex has the function of transporting ammonium and that a previously proposed mechanism of erythrocyte shape control is valid. PMID- 11918446 TI - Dynamic analysis of inter-words time intervals: a method to analyze the structure of communicative signals. AB - Speech analyses are usually focused on words as signifiers ignoring inter-words time intervals (IWIs), which are related to the 'form' of speech, rather than to its 'content'. Applying the method of power spectrum analysis to inter vocalizations time intervals of bird singing, underlying periodic processes were detected. In contrast, human IWIs revealed non-periodicity, which may be random or chaotic. To differentiate between these two possibilities, the non-linear dynamic methods of unstable periodic orbits and correlation dimension were applied to show that IWIs are characterized by a low dimensional chaotic attractor. Its correlation dimension of 3.2 +/- 1.1 suggests a minimum number of four variables underlying the system. The methods developed in the present communication can be further applied: (a) for the measurement of specific alterations in the processes underlying the form of speech in human disorders, i.e., schizophrenia, (b) for the assessment of normal and pathological developmental aspects of speech processes in children; (c) for comparing communicative signals between humans and other species. PMID- 11918447 TI - Body mass index: a measure for longevity. AB - Body mass index has important predictive value for mortality and morbidity both in normal subjects and in those suffering from particular pathologies. However, body mass index was introduced as a measure of body fat, which might not be expected to have such wide implications for various pathological conditions. We argue here that body mass index may actually be a measure for longevity. Our arguments are based on a well-established allometric scaling law for physiological time. The time between heart beats, the time between respirations, and longevity all scale as body weight to the 1/4 power in mammalian species ranging from shrews to blue whales. We find that body mass index also scales with body weight to the 1/4 power in humans from birth to one year of age, and again from approximately 5 to 17 years of age. On the assumption that in these two growth phases humans scale as do species, we postulate that body mass index is a measure of longevity. PMID- 11918448 TI - A comprehensive model for the aetiology of otitis media with effusion. AB - Otitis media with effusion is highly prevalent among young children. Adverse effects of this disorder are mainly restricted to the group of children with a history of recurrent or persistent otitis media with effusion. Early identification, assessment and intervention might prevent these adverse effects. Up to now it is not possible to distinguish these children from those with transient otitis media with effusion. This article presents a comprehensive model for the aetiology of otitis media with effusion. Eustachian tube functioning and the immunological response to environmental pathogens are the two core elements. This model can be used to formulate specific hypotheses about the interaction of several factors that may lead to the early identification of children who are likely to develop persistent or recurrent otitis media with effusion. PMID- 11918449 TI - Homocysteine levels in pregnant women who smoke cigarettes. PMID- 11918450 TI - Arsenic trioxide: a new immunomodulatory agent in the management of multiple myeloma. AB - Arsenic has been used as a medicinal for thousands of years. Several reports from China relative to its use mainly in acute promyelocytic leukemia, especially from the Shanghai group, has caused a resurgence in the investigation of the drug in the management of malignancies with focus on malignancies of hematologic origin. Arsenic is eliminated by many routes (urine, feces, sweat, milk, hair, skin, and lungs), although most is ultimately excreted in urine. Multiple myeloma is characterized by the clonal proliferation of malignant plasma cells in the bone marrow associated with bone loss, renal disease, and immunodeficiency. Preclinical evidence suggests an immunologic mechanism behind the therapeutic effects of As2O3 on myeloma cells. This appears to be achieved by a marked increase in lymphokine-activated killers mediated killing and up-modulation of CD38 and Cd54, two molecules involved in cell-cell interactions. Moreover, As2O3 alone or administered with ascorbic acid may provide a novel therapy for lymphoproliferative disorders. Preliminary clinical data in relapsed/refractory multiple myeloma suggest that As2O3 does have a role in the management of multiple myeloma; however, preclinical data show that the addition of ascorbic acid, and using As2O3 in combination with other active chemotherapeutic agents will enhance its role in managing the disease, and this is probably the position the drug will occupy in the armamentarium against myeloma. PMID- 11918452 TI - Treatment of refractory Hodgkin's disease with modified Stanford V program. AB - This study analyzes the results using an Stanford V modified program in the treatment of refractory Hodgkin's disease (RHD). We used cyclophosphamide instead of mechloretamine, and epirubicin instead of doxorubicin to avoid the risk of acute and late side effects associated with this drugs. Seventy-one patients with RHD were treated. All were at an advanced stage at therapy and had associated adverse prognostic factors. The complete response (CR) rate was 84% (60 patients; 95% confidence interval [CI]: 72-91%). At 5 yr, actuarial overall survival (CS) is 71% (95% Cl: 59-78%) and event-free survival (EFS) is 70% (95% CI: 59-79%). Only the duration of the initial complete response (> 12 mo) influenced the duration of EFS and OS. Toxicity was mild. Granulocyte colony-stimulating factor to ameliorate the presence of severe myelosuppression was used only in a few patients. Cardiac function was not affected and, until now, late side effects has not been observed. Thus, the use of this modified Stanford program retains the usefulness of the original scheme both with less frequent and less severe acute and late side effects. A controlled clinical trial in untreated patients comparing the Stanford program with standard chemotherapy is warranted to define the role of this therapeutic option in patients with Hodgkin's disease. PMID- 11918453 TI - Oral etoposide in patients with hematological malignancies: a clinical and pharmacokinetic study. AB - Tumor responses after daily oral administration of low-dose etoposide have been demonstrated in both hematological and solid tumors. The aim of the present phase II trial was to determine tumor response, and toxicity and to delineate the pharmacokinetics of oral low-dose etoposide in patients with hematological malignancies in a palliative treatment setting. Thirty-two patients with non Hodgkin's lymphoma (NHL), acute myeloblastic (AML) and lymphoblastic leukemia, multiple myeloma, and myelodysplastic syndrome (MDS) were included. Patients were given oral etoposide, 100 mg once daily for 14 d in a 21-d cycle. Serum etoposide concentrations were determined on d 1, 7, and 14 of every cycle before etoposide administration and, in addition, 1, 2, 3, 4, and 24 h after drug intake on d 1. The median age of patients was 68 yr (range: 50-89 yr). The median time from diagnosis to inclusion in the study was 21 mo (range: 0.5-144 mo) and most patients had advanced disease and were heavily pretreated. Eleven patients completed three or more cycles. Eight of 11 patients with acute leukemia and 1 of 2 with MDS received only 1 course because of toxicity (n = 5) or progression (n = 4). One patient with AML, a Jehovah's Witness, was treated up-front and achieved a complete remission and two patients with low-grade NHL gained a complete and a partial remission, respectively. Twenty-one of 32 patients were evaluable for toxicity during the first cycle. In 67%, the white blood cell count nadir was < 2.0 x 109/L and in 38% < 1.0 x 10(9)/L. Platelet count nadir was less than 25 x 10(9)/L in 24% of evaluable patients. During all cycles (n = 79), eight patients developed febrile neutropenia, four of whom with a fatal outcome. The correlation between the area under the curve (AUC) of the free fraction of etoposide and leukopenia was statistically significant at a log analysis (n = 12; p < 0.05). There was also a statistically significant correlation between the AUC and the 24 h concentration (n = 15; p < 0.005) and between the concentrations at 24 h and d 7 (n = 11; p < 0.005) of the free fractions of etoposide. In conclusion, etoposide had a moderate clinical effect in this group of heavily pretreated patients. Moreover, toxicity was substantial, in particular leukopenia, which correlated to the free-etoposide AUC. PMID- 11918454 TI - Successful treatment of gastrointestinal follicular lymphoma with rituxan and combination chemotherapy. AB - The clinical course of follicular lymphoma (FL) is well known. Although it is a chemosensitive disease, thereby allowing substantial palliation, recurrence is the rule; only a small subset who presents with limited stage disease is cured. Multiple attempts have been made over the past two decades to improve the survival of patients with FL, and a large number of phase III trials have been reported. These have included a variety of different therapeutic interventions, such as combination chemotherapy, recombinant interferons, new cytotoxic drugs, and immunologic agents. Most studies have not demonstrated that the use of a particular therapy convincingly prolongs survival. Follicular lymphoma cells express CD20 and are associated in most cases with the t(14:18) chromosomal translocation. Rituximab is a chimeric monoclonal antibody directed against the B cell CD20 antigen, which has been utilized for the therapy of B-cell non Hodgkin's lymphoma. Rituximab was shown to be active in FL, and studies of its effectiveness in combination with cytotoxic chemotherapy to increase the response rate are forthcoming. PMID- 11918455 TI - Hypereosinophilia and metastatic anaplastic carcinoma of unknown primary. AB - The differential diagnosis of eosinophilia may sometimes be difficult. Eosinophilia may occur in a diverse array of conditions from parasitic infestations to malignacies. Idiopathic hypereosinophilic syndrome has also been described. A 65-year-old male patient presenting with eosinophilia of obscure etiology is described in the present report. Three years after the diagnosis of eosinophilia, metastatic anaplastic carcinoma of unknown primary was detected. Differential diagnosis is disscussed briefly. It is stressed that patients with hypereosinophilia of unknown etiology must be screened for malignancy regularly during follow up. PMID- 11918451 TI - Treatment resistance of solid tumors: role of hypoxia and anemia. AB - Hypoxia is a characteristic property of locally advanced solid tumors, resulting from an imbalance between the supply and consumption of oxygen. Major pathogenetic mechanisms for the development of hypoxia are (1) structural and functional abnormalities of the tumor microvasculature, (2) increased diffusion distances, and (3) tumor-associated and therapy-induced anemia. The oxygenation status is independent of clinical tumor size, stage, grade, and histopathological type, but is affected by the hemoglobin level. Hypoxia is intensified in anemic patients, especially in tumors with low perfusion rates. Hypoxia and anemia (most probably via worsening of tumor hypoxia) can lead to therapeutic problems, as they make solid tumors resistant to sparsely ionizing radiation and some forms of chemotherapy. In addition to more direct mechanisms involved in the development of therapeutic resistance, there are also indirect machineries that can cause barriers to therapies. These include hypoxia-driven proteome and genome changes and clonal selection. These, in turn, can drive subsequent events that are known to further increase resistance to therapy (in addition to critically affecting long-term prognosis). Treatment resistance in anemic patients can be, at least partially, prevented or overcome by anemia correction, resulting in better locoregional tumor control and overall survival of patients. PMID- 11918456 TI - Elevated serum cancer antigen 125 levels in advanced abdominal tuberculosis. AB - A 48-yr-old female presented with a 1-yr history of pain in the hypochondrium and epigastrium. All routine investigations and computed tomography (CT) of the abdomen were done. CT findings revealed a well-defined cystic mass in the right ovary, and ascitis with features suggestive of secondaries over the omentum and peritoneal surface. The serum CA125 was 1255 U/mL (normal range 0-35 U/mL), which was indicative of ovarian malignancy. An exploratory laparotomy was performed. Histopathological examination of organs revealed the presence of granuloma. The patient was advised to undergo antitubercular treatment (ATT) and follow-up every month. After 1 mo of ATT, the CA125 level came down to 42 U/mL, which was near normal. As tuberculosis requires only a conservative management, we suggest that in cases of abdominopelvic mass with or without ascitis, high serum CA125 should always raise a suspicion of tuberculosis and a laparoscopy combined with peritoneal biopsy should be performed to confirm the diagnosis. This will prevent unnecessary laparotomies. Moreover, serum CA125 can be used to monitor the response of disease to antitubercular treatment. PMID- 11918457 TI - Primary malignant fibrous histiocytoma of the spleen and liver. AB - Malignant fibrous histiocytoma (MFH) is a distinct and pleomorphic form of sarcoma, which usually occurs in soft tissues but can be found in bones, kidney, larynx, lung, heart, and even aorta. Since the first description of MFH of the spleen by Govoni et al. in 1982, only 10 cases have been reported in the literature worldwide. We report on a 45-yr-old female with MFH of the spleen and liver, with special emphasizes on immunohistochemical findings. PMID- 11918459 TI - Genetics of epilepsy. AB - Understanding the molecular biology of epilepsy is a challenge for modern science. Epilepsy results from alternations in fundamental mechanisms of brain and membrane function. Although an understanding of the mode of inheritance and the etiology of genetic epilepsy syndromes forms the basis for genetic counseling, the development of specific therapies will come from knowing the basic mechanisms of epilepsy. Defining the genes causing epilepsy requires an unambiguous definition of seizure phenotype, along with the stability of that trait, an unremitting clinical course, and an abundance of clinical material. This article reviews the task of defining the genetics of epilepsy and discusses genetic methodology, idiopathic generalized and localization-related partial epilepsies, neuronal migration disorders, progressive myoclonus epilepsies, molecular biology of epileptogenesis, and future research. PMID- 11918458 TI - Unmet needs in pediatric epilepsy. AB - During the past 25 years, much new information about the epidemiology, pathophysiology, and prognosis of various forms of childhood epilepsy has been compiled. This has resulted in improved treatments that have helped an increasing number of children with epilepsy overcome unchecked seizures and the accompanying social stigma, enabling them to lead full and rewarding lives. The purpose of this supplement is to present a scientific and clinically relevant examination of some of the major breakthroughs in pediatric epilepsy, including a review of the latest advances in our understanding of the fundamental basis of childhood epilepsy. Additionally, this supplement will discuss persisting diagnostic issues, explore the role and use of new therapeutic options, and contemplate future treatment approaches. The articles are the result of a closed symposium held in January 2001. PMID- 11918460 TI - Diagnostic difficulty in infants and children. AB - Making an accurate diagnosis is the first and most critical step in the treatment of pediatric epilepsy, but it can be a daunting challenge for clinicians. Seizure types and syndromes in infants and very young children do not present with the same clarity and consistency as in adults. Work is currently being done to revise traditional International League Against Epilepsy classifications to make it easier for physicians to accurately diagnose and effectively treat epilepsy in infants and young children. A simplified approach can be formed by combining some of the International League Against Epilepsy recommendations with new findings on the clinical manifestations of pediatric seizures. Refinements in the new system will occur as knowledge of the pathophysiology of epilepsy and genetic susceptibilities expands, but even in its early stages the new classification system has much to offer, resulting in improved patient care and more effective management of the disease. PMID- 11918461 TI - Status epilepticus and acute serial seizures in children. AB - Status epilepticus is defined as a seizure that persists for a sufficient length of time or is repeated frequently enough to produce a fixed and enduring epileptic condition of 30 minutes or longer. Status epilepticus is a life threatening condition that often occurs in children. The degree of mortality and neurologic morbidity, as well as the risk for recurrence, is highly dependent on the etiology and duration of the seizures. Although much has been written about pediatric status epilepticus, many issues remain unresolved. A better understanding of the different types of seizures and their etiologies may help in the prevention and treatment of status epilepticus. The vast extent of status epilepticus in both children and adults mandates that new options for prevention and treatment be given a close scrutiny and high priority. This article will review the most current information on convulsive and nonconvulsive status epilepticus, including the potential for neurologic damage, changes in magnetic resonance imaging after status epilepticus, risk for recurrence, and current treatment options available for treating status epilepticus in children. PMID- 11918462 TI - Update on the epidemiology and prognosis of pediatric epilepsy. AB - Epilepsy is among the most common serious neurologic disorders in childhood. Epidemiologic studies over the past few decades have greatly increased current knowledge of the incidence and prognosis of seizures. Newer epidemiologic studies have used population- or community-based cohorts, and careful attention has been given to etiology and specific epilepsy syndromes, the two most important factors affecting prognosis. Risk of epilepsy is highest in patients with an associated serious neurologic abnormality, such as mental retardation or cerebral palsy. More than two thirds of patients with childhood-onset epilepsy ultimately achieve remission. Of those attaining remission on medications, approximately 70% remain seizure free when medications are discontinued. Mortality is increased in patients with epilepsy, but the increased mortality risk in childhood-onset epilepsy is primarily seen in patients with neurologic abnormalities or intractable epilepsy. Although long-term seizure outcomes are generally favorable, childhood-onset epilepsy is associated with adverse long-term psychosocial outcomes, even in patients attaining remission. This review summarizes recent data on the epidemiology and prognosis of pediatric epilepsy. PMID- 11918463 TI - Febrile seizures. AB - Febrile seizures are the most common form of childhood seizures, occurring in 2 to 5% of children in the United States. Most febrile seizures are considered simple, although those with focal onset, prolonged duration, or that occur more than once within the same febrile illness are considered complex. Risk factors for a first febrile seizure, recurrence of febrile seizures, and development of future epilepsy are identifiable and varied. Children with febrile seizures encounter little risk of mortality and morbidity and have no association with any detectable brain damage. Recurrence is possible, but only a small minority will go on to develop epilepsy. Although antiepileptic drugs can prevent recurrent febrile seizures, they do not alter the risk of subsequent epilepsy. This has led to a changing view of how we approach the treatment of these common and largely benign seizures. This chapter will review the current understanding of the prognosis and management of febrile seizures. PMID- 11918464 TI - Three new drugs for epilepsy: levetiracetam, oxcarbazepine, and zonisamide. AB - During the last decade, nine new antiepileptic drugs were approved for use by the US Food and Drug Administration. The characteristics of three of these new antiepileptic drugs-levetiracetam, oxcarbazepine, and zonisamide--are reviewed here. Their individual characteristics, including mechanism of action, efficacy, safety, and pharmacokinetics, are compared, and their effectiveness in treating specific seizure types is noted. As with all antiepileptic drugs, the efficacy, side-effect, and pharmacokinetic profiles must be matched to each individual's clinical profile to attain the maximum benefit. All three are unique and will be useful in expanding the aggregate of therapies available to clinicians treating diverse epilepsy syndromes and seizure types. This review focuses on results in adults; pediatric experience is reported in other articles in this supplement. PMID- 11918465 TI - Using the new antiepilepsy drugs in children. AB - New epilepsy treatment options are becoming available to physicians and may help provide safe and effective seizure management in patients for whom traditional drug therapies have been unsuccessful. Presented here is a brief discussion on the screening models used to test drugs for efficacy against various seizure types and the mechanisms of action and pharmacology of antiepilepsy drugs. This is followed by a more detailed review of the clinical pharmacology of the latest three antiepilepsy drugs introduced to the United States: levetiracetam, oxcarbazepine, and zonisamide. All three appear to have advantages over some traditional antiepilepsy drugs and with more clinical experience may replace them in certain seizure types or epilepsy syndromes. PMID- 11918466 TI - Pediatric partial and generalized seizures. AB - Eight new anticonvulsant medications have been approved in the United States since 1993, offering physicians a greater range of options for treating children with partial and generalized seizures. However, pediatric neurologists have been faced with limited pediatric pharmacokinetic and pharmacodynamic information. This article reviews the newer antiepilepsy drugs-gabapentin, felbamate, lamotrigine, topiramate, oxcarbazepine, levetiracetam, and zonisamide-and summarizes what is currently known about the safety and efficacy of these drugs in treating partial and generalized seizures in the pediatric population. Further studies are needed, however, to thoroughly evaluate their efficacy and safety in children. PMID- 11918467 TI - Lennox-Gastaut syndrome. AB - Lennox-Gastaut syndrome is a type of childhood epilepsy that has enormous detrimental effects on the patient's physical and developmental health and can also take a dramatic toll on the well-being of the patient's family. Lennox Gastaut syndrome is characterized by variable etiology, multiple types of intractable seizures, and cognitive impairment in most patients. It is one of the most difficult epilepsy syndromes to treat and is frequently resistant to treatment with standard antiepilepsy drugs. This article reviews the etiology of Lennox-Gastaut syndrome, characteristics of predominant seizure types, methods of evaluating patients for Lennox-Gastaut syndrome, and available treatments including antiepilepsy drug therapy, ketogenic diet, and surgical options. PMID- 11918469 TI - Progressive myoclonic epilepsies. AB - The progressive myoclonic epilepsies are a rare but extremely debilitating group of disorders that are difficult to diagnose and even harder to treat. They represent a heterogeneous subgroup of those with secondary generalized epilepsy. Efficacy of treatment is often measured in terms of slowing a patient's inevitable decline. Reviewed here are the classification of progressive myoclonic epilepsies, features of myoclonic seizures, the five most prevalent progressive myoclonic epilepsy syndromes-Unverricht-Lundborg disease, myoclonus epilepsy with ragged red fibers (MERRF) mitochondrial disease, Lafora's disease, neuronal ceroid lipofuscinoses, and sialidoses-and current treatment options. PMID- 11918468 TI - West's syndrome. AB - West's syndrome (infantile spasms) can be considered the classic disorder of the catastrophic childhood epilepsies. Although West's syndrome was identified 160 years ago, it is still not fully understood today. Because of the multiple etiologies associated with West's syndrome, the evaluation and treatment of the disease are complex, and, to date, there are no drugs approved by the US Food and Drug Administration specifically for the treatment of infantile spasms. The multiple etiologies associated with infantile spasms require that physicians take a broad look at all therapeutic options. There are, however, logical sequences that physicians should pursue when trying to treat this disorder. This article reviews the etiologies associated with West's syndrome, antiepilepsy drug therapies currently being used to treat the syndrome, and other treatment options available to clinicians. PMID- 11918470 TI - Advancing the medical management of epilepsy: disease modification and pharmacogenetics. AB - Despite the recent development of new antiepilepsy drugs, a significant number of children are still unable to achieve seizure freedom without side effects. Understanding the factors behind individual variability in antiepilepsy drug tolerability and dose response and incorporating these factors into a treatment plan would represent an important advance in epilepsy pharmacotherapy. A more thorough understanding of the epileptogenic process may allow clinicians to select antiepilepsy drugs that interrupt or modify various steps in the epileptogenic progression (ie, disease modification). Additionally, advances in the understanding of human genetics may allow for selection of antiepilepsy drugs and dosage regimens based on a patient's clinical characteristics and genotype (ie, pharmacogenetics). This article focuses on these two areas of potential improvement in the medical treatment of patients with epilepsy. Such methods of tailoring antiepilepsy drug therapy would be preferable to the trial-and-error system that is currently used. PMID- 11918471 TI - Vitamins in Spanish food patterns: the eVe Study. AB - OBJECTIVE: To describe vitamin intakes in Spanish food patterns, identify groups at risk for inadequacy and determine conditioning factors that may influence this situation. DESIGN: Pooled-analysis of eight cross-sectional regional nutrition surveys. SUBJECTS: Ten thousand two hundred and eight free-living subjects (4728 men, 5480 women) aged 25-60 years. Respondents of population nutritional surveys carried out in eight Spanish regions (Alicante, Andalucia, Balearic Islands, Canary Islands, Catalunya, Galicia, Madrid and Basque Country) from 1990 to 1998. The samples were pooled together and weighted to build a national random sample. METHODS: Dietary assessment by means of repeated 24-hour recall using photograph models to estimate portion size. Adjusted data for intra-individual variation were used to estimate the prevalence of inadequate intake. A Diet Quality Score (DQS) was computed considering the risk for inadequate intake for folate, vitamin C, vitamin A and vitamin E. DQS scores vary between 0 (good) and 4 (very poor). Influence of lifestyle (smoking, alcohol consumption and physical activity) was considered as well. RESULTS: Inadequate intakes (<2/3 Recommended Dietary Intake) were estimated in more than 10% of the sample for riboflavin (in men), folate (in women), vitamin C, vitamin A, vitamin D and vitamin E. More than 35% of the sample had diets classified as poor quality or very poor quality. Factors identified to have an influence on a poor-quality diet were old age, low education level and low socio-economical level. A sedentary lifestyle, smoking, usual consumption of alcohol and being overweight were conditioning factors for a poor-quality diet as well. CONCLUSION: Results from The eVe Study suggest that a high proportion of the Spanish population has inadequate intakes for at least one nutrient and nearly 50% should adjust their usual food pattern towards a more nutrient-dense, healthier diet. PMID- 11918472 TI - Vitamin status in different groups of the Spanish population: a meta-analysis of national studies performed between 1990 and 1999. AB - OBJECTIVE: Studies performed on the nutritional status of the Spanish population have been very heterogeneous with respect to methodology, sample size, geographical location, socio-economic level and health status of the subjects involved. In order to gain an overall view of the state of knowledge in this area, a meta-analysis was performed on the results of all such studies undertaken in Spain between 1990 and 1999. RESULTS: The dietetic data reviewed showed mean intakes similar to those recommended. The results obtained for the biochemical parameters analysed were within normal limits. However, results for some vitamins were very close to the lower established limits for the reference intervals (recommended intakes or blood levels), meaning that a variable percentage of subjects show values lower than those recommended. The percentage of subjects with intakes below those recommended for niacin, vitamin B12 and C was small. The percentage of inadequate intakes of thiamine and riboflavin was small as well. However, for all other vitamins, especially D and E, the number of people with intakes below recommended was rather high, particularly so in some studies. At blood level, deficiencies of vitamins B12, A and E were infrequent. However, for all other vitamins, prevalence of deficiency varied within a wide range. With respect to vitamins D, B1, B2 and B6, over 50% of the population showed inadequate levels. CONCLUSIONS: The methodologies used in the studies included in this review were very varied and the knowledge gained is still incomplete. Despite the average Spanish diet often being regarded as satisfactory, this review and other studies show the situation can be improved. PMID- 11918473 TI - Fortified foods. Criteria for vitamin supplementation in Spain. AB - OBJECTIVE: To review and analyse criteria for vitamin supplementation and fortification in Spain. DESIGN: Systematic review of scientific literature and simulation analysis of food fortification. METHODS: A simulation analysis using a fortified beverage was performed in a random sample of 2855 children aged 2 to 24 years in Spain. RESULTS: High-risk groups for vitamin supplementation and fortification in Spain are highlighted, and target vitamins considered have been: folic acid, vitamin A, vitamin E, vitamin D and vitamin B12 (particularly in the elderly). A beverage fortified with vitamins C, A, B1 and B6 may contribute to improving the intake of all of these vitamins with the exception of vitamin A, since the Recommended Nutrient Intake is already covered with current consumption. CONCLUSIONS: A detailed knowledge of nutritional status helps to ensure the rationale and follow up of nutrient supplementation and fortification. PMID- 11918474 TI - Early nutrition and risk of disease in the adult. AB - The differentiation of key metabolic systems that occurs during intrauterine life is greatly influenced by environmental nutritional conditions, which in turn are related to maternal nutritional status. In postnatal life, childhood exposure to slow-acting environmental factors, primarily through the diet, will begin to condition adult susceptibility to diseases. Examples of these dietary factors are intake of saturated fat, sodium, calcium, etc. For example, bone calcium accumulation during childhood and adolescence will be a major determinant of risk of osteoporosis later in life. Similarly, a high intake of saturated fat during childhood may promote the process of atherosclerosis in persons with genetic susceptibility, thus accelerating the clinical manifestations of coronary heart disease in adult life. These findings, although still not completely clarified, constitute a significant opportunity for preventive intervention. While preventive intervention in adult life may reduce risk, this is usually difficult and results are often limited. One example would be obesity. In contrast, interventions early in life, aimed at reducing these early risk factors, could potentially result in major reductions in the incidence of several diseases of adults. PMID- 11918475 TI - School health services and community nutrition: a historical perspective. AB - The Bilbao School Health Service was created at the beginning of the century with the aim of preventing transmittable diseases among children as well as improving nutrition. At that time such services were established in many other countries. Since then, according to evolving societal changes and emerging needs, the Service has reoriented its scope and structure towards the Health Promotion scheme. Current tasks include health screening examinations and hygiene surveillance as well as preventive and health education programmes. PMID- 11918476 TI - School meals in Catalonia: surveillance and quality control. AB - The Education Council of the Catalonian government drew up a programme to improve the quality control of school canteens in Catalonia. The objectives of this programme were to contribute to developing healthier eating habits in school children through school canteens and to improve school canteen services considering other aspects such as service, operation, management, end-users, frontline staff and supervisors of menu planning. This paper deals with two programme components aiming at introducing Nutrition in the school syllabus and school menu assessment. Various departments, organisations and associations were involved in programme development and implementation. Initial analysis was based on information collected from formal administrative reports completed by a survey on 100 primary schools. From this preliminary analysis it was concluded that problem identification needs to be completed by an understanding of underlying reasons in order to design adequate solutions and guarantee implementation. PMID- 11918477 TI - Dietary guidelines for pregnant women. AB - The nutrition of pregnant women is decisive in the course of gestation and the health of both mother and child. However, in the Spanish population, clinical monitoring of nutrition is rare, the control of bodyweight receiving most attention. Many studies show that pregnant women take too much fat and too little carbohydrate, and that intakes of fibre and several trace elements are lower than recommended. Although many people think they know what a correct (varied and balanced) diet is, the concept is often misleading. Generally, they do not match the ideal theoretical framework for achieving an adequate energy profile of the diet. Neither do they facilitate intakes of vitamins and minerals close to recommended levels. Nutrition education programmes are therefore required to explain adequate dietary guidelines to pregnant women, and indeed to all women of childbearing age. Considering the criteria suggested by a number of researchers concerning the number of food portions from each food group that pregnant women should include in their diets, the following guidelines for daily consumption are proposed: 3-4 portions of milk products, 2-3 portions of meat, fish or eggs, and 3 portions of fruit, 4-5 portions of vegetables or greens and 7-8 portions of cereals and legumes (a portion is defined as the amount of food that would be found on an average plateful or the normal units of consumption of a food). PMID- 11918478 TI - Breast-feeding in Spain. AB - Human milk is the best way to nurture the human infant. By breast-feeding their babies, mothers provide them with the best opportunities to wholly develop their potential, while protecting the infants and themselves from a whole range of diseases in the near future and in the years to come. Even though these benefits are widely known and there is ample scientific evidence on the topic, it seems from published data that Spanish women are not breast-feeding their babies as much and for as long as they should. Less than 90% start breast-feeding, at 1 month there is already an attrition of 30%, at 3 months more than half of the infants are taking artificial milk and by 6 months only 10% continue to breast feed their infants. Low birth weight, Caesarean section and low study level are among the more significant factors that negatively affect breast-feeding. There is still work to do to improve this situation. Promotion of breast-feeding among the general population, mothers and health professionals is needed. PMID- 11918479 TI - Breast-feeding in Europe--rationale and prevalence, challenges and possibilities for promotion. AB - The status reports and other information collected showed that interpretation of the data on breast-feeding prevalence and duration collected at national or regional level within European countries is difficult, since this information is not collected in every country or it is gathered under different criteria. However, there seem to be vast differences in prevalence of breast-fed children and breast-feeding duration between European countries and possibly within countries. There is a need to establish monitoring systems enabling comparability of data between countries. Assessing determinants for breast-feeding is required as well. There are a number of important consensus documents supporting breast feeding action. These documents are related to either one or more of the following categories: health benefits of breast-feeding; recommendations regarding breast-feeding duration and exclusiveness; providing guidance on breast feeding promotion. Current recommendation is exclusive breast-feeding for 6 months. PMID- 11918480 TI - Dietary guidelines for the breast-feeding woman. AB - Sociodemographic and economic changes in Spain have played a role in the choice of infant feeding as more and more women enter the workforce. Nevertheless, the prevalence of breast-fed-only infants at 6 weeks remains fairly high, at 65.5% in 1999, although there is a sharp decline by 3 and 6 months postpartum. Achieving adequate nutritional status for lactation principally begins during pregnancy, as this constitutes the biological stage for accumulating nutrient stores. The benefits of breast-feeding for the mother and infant are presented, along with nutrition and dietary guidelines for the nursing woman. Apart from dietary considerations, special attention should be paid to encouraging a positive attitude and environment for breast-feeding success. PMID- 11918481 TI - Preventive nutritional supplementation throughout the reproductive life cycle. AB - Current interest in preventive nutritional supplementation strategies during the reproductive cycle in developed countries focuses mainly on iron and folic acid. Iron deficiency is the most common cause of anaemia. Inadequate iron nutrition is mainly a problem in developing countries but it also seems to affect a large proportion of pregnant women in the industrialised world. There is no chance for diet modification alone to meet iron requirements during pregnancy. However, dietary advice must be provided to increase iron intake and iron bioavailability. The best choice is prescribed supplementation adapted to the individual's needs. Folic acid supplementation during the periconceptional period reduces the occurrence and recurrence of neural tube defects (NTDs). According to the updated edition of American Dietary Reference Intakes, recommended dietary allowance for folate for women of childbearing age varies throughout the reproductive cycle from 400 microg day(-1) of dietary folate equivalents (DFEs) for non-pregnant women, to 600 microg day(-1) DFEs for pregnant women and 500 microg day(-1) DFEs in lactating women. Increasing dietary folate, fortifying staple foods with folic acid or use of folic acid supplements can increase folate intake. PMID- 11918482 TI - Food services for the homeless in Spain: Caritas Programme for the Homeless. AB - Homelessness includes a wide range of people, from those living on the street (strictly homeless) to people living temporarily with relatives or friends because they have lost their home. Programmes for the homeless should design strategies aimed at encouraging insertion processes, thus considering the different dimensions involved. Within this framework Caritas delivers an array of assistance services for the homeless facing the stages of insertion. In Spain there are 315 centres offering meals for 20000 people, most of them along with other services such as reception, residential premises or day-care centres. Caritas is responsible for 42% of them. Overall, it is easier to get a lunch than any other meal during the day, followed by dinner, while it is difficult to get a breakfast. Social dining rooms are valuable places for a first contact (first stage of the insertion process) and even to start the second stage (personal recovery). PMID- 11918483 TI - Social inequalities and the burden of food-related ill-health. AB - Increased health inequalities are a result of poverty and social inequalities. Examples of how food intake is affected by poverty exist throughout the European Region. The diets of low-income groups are likely to be inadequate. Low-income groups and specific groups such as children, adolescents, pregnant and lactating women, and older people often face problems gaining access to a healthy variety of safe foods. Safe healthy food may not be accessible to those most in need. Yet access to a safe and varied healthy diet is a fundamental human right and policies are needed to address this. PMID- 11918484 TI - Screening for malnutrition in dwelling elderly. AB - The health of elderly people is often affected by inadequate nutritional intake. Psychosocial determinants and the higher prevalence of acute and chronic illnesses are risk factors for nutritional deficiencies in the elderly. Nutritional assessment (NA) or nutritional risk screening must be an important element in care programmes for elderly people in the community and should be part of all geriatric assessment programmes. PMID- 11918485 TI - Guidelines for nutrition support in the elderly. AB - Nutritional support in the elderly not only co-operates in pharmacological treatment but also very often is a primary therapy for their health. The type of artificial nutrition (AN) to use will depend on the present illness and the previous health record. Due to the fact that enteral feeding (EF) is less expensive and aggressive we should use EF whenever possible, leaving parenteral nutrition (PN) for specific situations where EF should not be used. AN, if properly prescribed, formulated, administered and monitored, is safe as long as qualified personnel are trained in its use. Combined AN (oral, enteral and parenteral) allows a step-by-step improvement that could lead to final oral feeding. Finally, while it is true that age should not be considered in isolation as a contraindication for AN, we should be aware that, in final life stages, oral feeding can be the only satisfaction left for the elderly. PMID- 11918486 TI - Nutrition and ageing. AB - The reviewed literature indicates that, even in industrialised countries, the nutrition of mature and aged subjects is often inadequate (because of deficiency or excess), which may lead to premature or pathological senescence. Recent nutritional research on ageing laboratory animals shows that dietary restriction may be the most effective procedure to achieve a long and disease-free life span, probably owing to a better protection against mitochondria-linked oxygen stress. Likewise, the experimental and clinical work from many laboratories, including our own, indicates that age-dependent changes in the cardiovascular and immune systems are linked to oxygen stress and that an adequate intake of dietary antioxidants may protect those systems against chronic degenerative syndromes in the physiopathology of which reactive oxygen species (ROS) play a key role. The extant data indicate that the antioxidant vitamins C and E are centrally involved in defending the above two systems against ROS attack. Moreover, recent research suggests that the glutathione-related thiolic antioxidants, thiazolidine carboxylic acid (thioproline) and N-acetylcysteine, as well as the phenolic liposoluble 'co-antioxidants' of Curcuma longa, may have a significant protective effect against age-related atherogenesis and immune dysfunction. Key messages from this paper are the following. (1) It is generally accepted that oxygen free radicals released in metabolic reactions play a key role in the physiopathology of 'normal ageing' and of many age-related degenerative diseases. (2) Consumption of adequate levels of antioxidants in the diet is essential in order to preserve health in old age. (3) A certain degree of protection against atherogenesis and immune dysfunction may be achieved by preventing vitamin E deficiency and an excessive oxidation of the glutathione-supported thiol pool. PMID- 11918487 TI - A European Master's Programme in Public Health Nutrition. AB - Effective population-based strategies require people trained and competent in the discipline of Public Health Nutrition. Since 1997, a European Master's Programme in Public Health Nutrition has been undergoing planning and implementation, by establishing initial quality assurance systems with the aid of funding from the European Commission (DG SANCO/F3). Partners from 17 European countries have been involved in the process. A European Network of Public Health Nutrition has been developed and accredited by the European Commission. PMID- 11918488 TI - Developing an evidence-based approach to Public Health Nutrition: translating evidence into policy. AB - The aim of this paper is to highlight the importance of an evidence-based approach to the development, implementation and evaluation of policies aimed at improving nutrition-related health in the population. Public Health Nutrition was established to realise a population-level approach to the prevention of the major nutrition-related health problems world-wide. The scope is broad and integrates activity from local, national, regional and international levels. The aim is to inform and develop coherent and effective policies that address the key rate limiting steps critical to improving nutrition-related public health. This paper sets out the rationale for an evidence-based approach to Public Health Nutrition developed under the umbrella of the European Network for Public Health Nutrition. PMID- 11918490 TI - Dietary guidelines for the Spanish population. AB - Therapeutic properties have been attributed to the diet since ancient times. Epidemiological evidence starting in the early 1960s supports the relationship between different dietary components, dietary balance and the development of risk factors for prevalent chronic diseases in Western societies. There is also evidence supporting the protective effect of other components of the diet and lifestyle, like regular physical activity. During the last two years, groups of Spanish experts in the field of food, nutrition and health sciences have been involved in the process of analysing each food group in Spanish food patterns under a standard format. For each food group, current level of consumption, trends and scientific evidence linking each particular group or its components with patterns of disease, risk factors or protective effects have been determined from a public health perspective. Conclusions and recommendations for each food group, agreed under consensus by the Spanish Society of Community Nutrition (SENC) collaborative group of experts, are summarised and reflected in the Food Guide Pyramid presented in this paper. PMID- 11918489 TI - Spanish food patterns. AB - Data from household food budgetary surveys and regional, population-based, cross sectional studies show a declining trend in energy intake in the last decades, also applicable to protein, fat and carbohydrate intakes in absolute terms. The pattern suggests a lower fat content in the diet. Protein intake is 200% of the recommended level. Spanish food patterns underwent dramatic changes between the 1960s and the 1980s, including an important increase in fat intake, up to 44% energy intake from fat in the 1980s. The trend changed in the 1990s, and current fat supply provides an average of 37% of energy intake. Main sources of fat in the Spanish diet are added fats for cooking (49% of total fat intake), especially olive oil; meat and meat products (25%); and a lower percentage provided by milk and dairy products. Most saturated fatty acids are supplied by meats (30%) and dairy products (27%). Rapid urbanisation processes and the growing proportion of females in the active workforce have led to important changes in food patterns in the last decades. On the one hand, some changes had a positive impact, such as increasing variety in the diet and access to food; on the other hand, some changes moved the Spanish diet away from the traditional Mediterranean food pattern. Current food patterns evidence high consumption of animal products: meat, fish, milk and dairy products. It seems advisable to increase consumption of plant foods, particularly whole-grain cereals, potatoes and pulses. Intake of fruit and vegetables shows a shift towards a greater consumption of processed foods rather than fresh products, and overall a greater consumption would be recommended. PMID- 11918491 TI - Nutritional objectives for the Spanish population. Consensus from the Spanish Society of Community Nutrition. AB - The objective of this paper is to present the development of the Nutritional Objectives for the Spanish Population. Preparation of draft documents contributed by different working groups was followed by a consensus meeting held in Bilbao on 5-7 October 2000, hosted by the Spanish Society of Community Nutrition and sponsored by the World Health Organisation. Establishing nutritional guidelines was conducted by: (1) analysing current food and nutritional data from nutritional surveys, for intermediate objectives; and (2) reviewing current scientific knowledge for final objectives. The objectives include intermediate and ultimate figures, and comprise percentage of energy from macronutrients and fatty acids, fruit and vegetable consumption, frequency of sweets, physical activity and body mass index, folate, calcium, sodium, fluoride and iodine intake, dietary fibre, cholesterol, alcohol and duration of breast-feeding. The nutritional objectives for the Spanish population create a rational framework for the development of dietary guidelines and nutritional policies in Spain, within a Mediterranean context. PMID- 11918492 TI - Nutrition intervention strategies in chronically malnourished regions: preventing endemic goitre in Togo. AB - Iodine deficiency disorders (IDDs) are an important problem in the world. Some 760 million people suffer from goitre and 1600 million are at risk. In Togo, a small West African country, the prevalence of visible goitre in 6-12-year-old children was 21.6% and 5.3% in two endemic regions in 1999. Goitre is considered endemic as it affects one adult in five. UNICEF-Togo (United Nations Children's Fund) in co-operation with the Togolese government has implemented different programmes with the aim of improving the survival and development of children and women and to defend and promote children's rights. The aim of this paper is to describe the procedures followed and key results of a school-based nutrition education project implemented in Togo to prevent iodine deficiency disorders, by encouraging use of iodised salt. PMID- 11918493 TI - Nutrition education training of health workers and other field staff to support chronically deprived communities. AB - This paper focuses on the provision of adequate training in nutrition education to health and other community development workers for their improved performance and achievement. The difficulties encountered and special care needed when dealing with low-income, chronically deprived communities are raised. A brief analysis of past and present trends in nutrition education is presented to show the progress made from restricted, authoritative and not very successful proposals to more comprehensive and participatory approaches. The need to train and update regional and field-level personnel on the new approaches, theories and models offered by nutrition education is highlighted, but the scant availability of resources for training activities may be a great limitation for this undertaking. The contribution of educational, social, psychological and communication sciences, as well as marketing, in improving and broadening the performance of health and nutrition education is recognised. Some successful nutrition education projects, implemented in different regions, using various approaches, have managed to improve the nutrition situation of low-income groups and could be used as good examples to be followed. Recommendations for implementing nutrition education projects or activities need to consider some prerequisites, such as good knowledge and analysis of the nutrition situation, careful selection of the strategies and methods, careful planning and implementation, and clear definition of the procedures and instruments for follow up and evaluation. PMID- 11918494 TI - Food transitions in the 20th-21st century. AB - Food transitions in history have been fast, violent processes in some cases, while on other occasions they have taken place slowly, overlapping the traditional food culture as food items are replaced by new ones. Despite the contribution of potato and maize to avoiding famine in Europe, the exclusive crop system caused serious problems. Main food transitions are currently taking place in Asia. Emerging economies face the dilemma of improving the health and well being of the population while avoiding excess and surplus. The challenge is twofold: on the one hand, to counteract the negative effects of food transitions and, on the other, to fight against hunger and malnutrition. PMID- 11918495 TI - Breakfast and performance. AB - Evidence suggests that the effect of fasting on performance is not uniform, but it is dependent on the basal nutritional status of the subject. Breakfast consumption has a short-term effect in improving selected learning skills, especially work memory. School breakfast programmes have a positive effect on the nutritional status of children, on school attendance and probably on dropout rates. The effect of breakfast consumption on school performance depends on the interaction between the programme, student characteristics (malnutrition) and school organisation. Unless the school setting guarantees a minimum quality standard, the benefits of breakfast consumption will not be evident in performance in complex areas like language or maths. PMID- 11918496 TI - Food patterns of Spanish schoolchildren and adolescents: The enKid Study. AB - OBJECTIVE: To evaluate dietary habits and nutritional status of Spanish schoolchildren and adolescents. DESIGN: Cross-sectional study by face-to-face interview. METHODS: A random sample of 3534 people aged 2 to 24 years were interviewed by a team of 43 dietitians in the subjects' homes. Interviews included 24-hour recalls (a second 24-hour recall in 30% of the sample), a food frequency questionnaire and other questions including lifestyles, knowledge and food preferences. RESULTS: The participation rate was 68%. Mean daily energy consumption was 2189 kcal among males and 1781 kcal among females, and the percentage of energy from fat and saturated fat was 39.8% and 13.4%, respectively, without any differences by gender. Of females, 95% showed folic acid intakes below 200 microg day(-1). Consumption expressed as food groups reflected a westernised Mediterranean pattern. CONCLUSIONS: The enKid Study provides reliable and useful nutrition information for children and adolescents in Spain, having incorporated a rigorous methodological design within a representative sample. PMID- 11918497 TI - Breakfast consumption in Spanish children and young people. AB - BACKGROUND AND OBJECTIVES: Breakfast consumption is important for nutritional balance in all population groups. The objective of this paper is to describe breakfast consumption patterns in Spanish children and young people. DESIGN: Random population cross-sectional nutritional survey. METHODS: A random sample of 3534 people aged 2 to 24 years were interviewed by a team of 43 dietitians. The protocol included 24-hour recalls (a second 24-hour recall in 30% of the sample), a food-frequency questionnaire and other questions including lifestyles, knowledge and food preferences. RESULTS: Participation rate in the survey was 68%. Of the boys, 91.2% and 92.2% of the girls in the sample usually had breakfast. Males aged 18 years and above and 14-17-year-old girls showed the lowest consumption rates. Some 4.1% of the sample did not have any food at all before lunch. Main sources of energy with breakfast were milk and dairy products (34%) and buns, cookies, croissants, etc. (24.6%). Some 44.7% of the sample was classified in the poor-quality breakfast group. Subjects from a low socio economic background and low educated parents had lower Quality Breakfast Score. CONCLUSIONS: The proportion of children having breakfast before attending school has increased in Spain in recent decades. However, intake of breakfast is not adequate in a large proportion of this population and a number of children and young people still do not eat anything before lunch. School-based nutrition education involving families, teachers and health professionals, supported by community actions, can contribute to improve this situation. PMID- 11918498 TI - Helping to promote healthy diets and lifestyles: the role of the food industry. AB - In order to be successful, public health nutrition strategies require the active collaboration of all stakeholders in the promotion of healthy diet and lifestyle patterns. The food industry plays an important role both in providing products that meet consumers' needs in terms of taste, convenience, quality, nutrition and value as well as in communicating to consumers about the importance of good nutrition, including the contribution of specific foods to a balanced diet. The food industry contributes to educational efforts regarding healthy diets and lifestyles both directly--through product labelling, advertising, educational materials, on-line communications and information provided by Consumer Services departments--and indirectly, through active involvement and participation in educational programmes pursued in collaboration with nutrition and health education authorities. Through ongoing dialogue with its consumers and research conducted on consumer knowledge and attitudes towards diet, the food industry can ensure that communications developed are motivating and relevant to consumers' lives. In this paper, the specific contribution of the food industry will be illustrated through the promotion of healthy eating habits among children, focusing in particular on the importance of the breakfast meal. PMID- 11918499 TI - Relationship between asthma drug therapy patterns and healthcare utilization. AB - BACKGROUND AND OBJECTIVE: Asthma drug therapy problems contribute significantly to preventable hospitalizations and increased healthcare use in asthmatics. Since asthma patients often require >1 medication for control of symptoms, concurrent asthma drug therapies may be important in predicting excessive healthcare utilization. The purpose of this study was to link inappropriate asthma drug therapy patterns and selected patient demographics to healthcare utilization. METHODS: This study was a retrospective, cross-sectional analysis of Ohio Medicaid medical, institutional, and prescription claims. We included ambulatory patients aged 15-65 years who had > or =2 claims for asthma (493.x) and who were continuously enrolled in the Medicaid fee-for-service program for the 12-month period from April 1998 through March 1999. We examined age, race, gender, metropolitan residence, presence of gastroesophageal reflux disease, and the usage patterns of inhaled corticosteroids, short-acting beta(2)-agonists, long acting beta(2)-agonists, theophylline, and leukotriene receptor modifiers to identify asthma drug therapy problems based on national guidelines. The primary outcomes included the number of asthma-related hospitalizations, asthma-related emergency department visits, and oral steroid bursts. RESULTS: Among 10 959 asthma patients, only 46.8% of the study patients received >1 puff of inhaled corticosteroid per day. Forty-four percent of the patients received >3 puffs of short-acting beta(2)-agonists per day. The most common outcome was an oral steroid burst (46.5%). Patients on high doses of short-acting beta(2)-agonists had the greatest odds of receiving an oral steroid burst and were most likely to be hospitalized. African Americans were more likely to incur a hospitalization or emergency department visit. Women had greater odds of any undesirable asthma outcome. Higher use of short-acting beta(2)-agonists led to higher odds of receiving a steroid burst or being hospitalized. Leukotriene receptor modifier use was related to higher levels of all outcomes. CONCLUSIONS: A large percentage of Ohio Medicaid patients were not receiving asthma medications in compliance with the National Heart, Lung, and Blood Institute guidelines. Despite nearly a decade of national efforts, asthma drug therapy patterns still have substantial room for improvement and continue to be associated with excess healthcare utilization. PMID- 11918500 TI - Intravenous versus oral corticosteroids for treatment of acute asthma exacerbations. AB - OBJECTIVE: To compare the duration of hospitalization of patients treated with either oral or intravenous corticosteroids for an acute asthma exacerbation. METHODS: A retrospective chart review was performed on a random sample of inpatients. Patients were included with the following: a discharge diagnosis of an acute asthma exacerbation, a past medical history significant for asthma, age between 16 and 60 years, and treatment with either oral or intravenous corticosteroids at the time of admission. Exclusion criteria included: patients receiving chronic prednisone therapy, a past medical history significant for chronic obstructive pulmonary disease, an admission to the intensive care unit, or a consistent smoking habit of at least 1 pack daily. Length of hospitalization was the primary outcome measured. Secondary outcomes included 24-hour peak expiratory flow rate, 24-hour pulse oximetry (pO(2)), and amount of beta-agonist and ipratropium used. RESULTS: Fifty-three patients were included in the final data analysis. Patients were grouped by route of corticosteroid administration (intravenous or oral). No significant differences were noted between the 2 groups for race, gender, age, height, weight, admission peak expiratory flow rate, admission pO(2), or types of asthma medications used prior to admission. No significant differences were demonstrated in any of the outcome measures. CONCLUSIONS: Both the intravenous and oral corticosteroid groups demonstrated similar clinical outcomes and lengths of hospitalization in the treatment of acute asthma exacerbations. These results support the initial use of oral corticosteroids for the treatment of acute asthma exacerbations in adult patients admitted to a general medical service. PMID- 11918501 TI - Pharmacy-based bone mass measurement to assess osteoporosis risk. AB - OBJECTIVE: To evaluate elderly women's knowledge of their skeletal status, assess adequacy of calcium intake, determine the prevalence of low bone density, and determine whether peripheral bone density testing led to medical interventions in a group of rural, elderly Wisconsin women recruited in community pharmacies. DESIGN: Recruiting notices were posted in each pharmacy, and eligible women were enrolled in the order in which they volunteered. Each completed a fracture-risk questionnaire. Calcaneal bone density was measured within the following 6 weeks, using peripheral dual-energy X-ray absorptiometry. Mail surveys were used to assess interventions subsequent to the womens' study participation. SETTING: The study was conducted at 5 community pharmacies in rural Wisconsin. RESULTS: Of 133 women, 20% had calcaneal osteoporosis, defined as a T score < or =2.5 (calcaneal bone density <2.5 SDs below the young reference database). Thirty percent of women met National Osteoporosis Foundation (NOF) treatment criteria based on heel bone density and NOF-designated risk factors. Of those meeting treatment criteria, 75% were unaware of their low bone mass. Half of the women received <1200 mg/d of calcium, the recommended dose for osteoporosis prevention. Those who were taking a calcium supplement were much more likely to receive the recommended amount. Women who had discussed bone density test results with their physicians were more likely to receive central dual energy X-ray absorptiometry (DXA) measurements and/or start antiresorptive therapy than women who did not. CONCLUSIONS: Rural, elderly Wisconsin women are at substantial risk for osteoporosis, based on calcaneal bone density, but most are unaware of their risk. Compounding this risk is low calcium intake. Community screening of rural, elderly women by peripheral bone density measurement can lead to medical interventions in such individuals. PMID- 11918502 TI - Discontinuing or switching selective serotonin-reuptake inhibitors. AB - OBJECTIVE: To describe reasons for discontinuing or switching selective serotonin reuptake inhibitors (SSRIs) at 3 and 6 months after starting treatment, and to identify information provided to patients that may help prevent premature discontinuation of medication. METHODS: Telephone surveys were conducted at 3 and 6 months after patients (n = 672) were started on an SSRI for a new or recurrent case of depression. RESULTS: Significantly more patients discontinued or switched their SSRI because of an adverse effect within the first 3 months of starting (43%) compared with the second 3 months (27%; p = 0.023). The adverse effect most frequently reported as the reason for early discontinuation or switching was drowsiness/fatigue (10.2%), followed by anxiety, headache, and nausea - all at just over 5%. The odds ratio for discontinuation was 61% less in patients who recalled being told to take the medication for at least 6 months compared with those who did not (OR 0.39; p < 0.001). Patients who recalled being informed of potential adverse effects increased their reported incidence of mild to moderate adverse effects by 55% (OR 1.55; p < 0.05) without affecting rates of premature discontinuation (OR 1.06; p = 0.77). CONCLUSIONS: Adverse effects are the most frequent reason for discontinuing or switching SSRIs within the first 3 months of treatment. Patients are more likely to continue taking their antidepressant if they fully understand how long to take the medication. Informing patients of potential adverse effects does not appear to prevent premature discontinuation, but may increase the patient's awareness and reporting of mild to moderate adverse effects. PMID- 11918503 TI - Collaborative care model to improve outcomes in major depression. AB - OBJECTIVE: To develop a pharmacist intervention to improve depression care and outcomes within a primary care setting. METHODS: Pragmatic, randomized trial of a clinical pharmacist collaborative care intervention versus usual care in a busy, academic family practice clinic. RESULTS: Seventy-four patients diagnosed with a new episode of major depression and started on antidepressant medications were randomized to enhanced care (EC) or usual care (UC) groups. EC consists of a clinical pharmacist collaborating with primary care providers (PCPs) to facilitate education, initiation, and titration of acute-phase antidepressant treatment to monitor treatment adherence and to prevent relapse. Control patients receive UC by their PCP. The main end point is reduction of depression symptoms over time as measured by the Hopkins Symptom Checklist (SCL-20). Other outcomes include the Diagnostic and Statistical Manual of Mental Disorders, (DSM-IV) criteria for major depression, health-related quality of life measured by the Medical Outcomes Study Short Form 12 (SF-12), medication adherence, patient satisfaction, and healthcare utilization. The main end point and the cost of treating major depression will be used to estimate the cost-effectiveness of the collaborative care model. CONCLUSIONS: The study is a unique, ongoing trial that may have important implications for the treatment of depression in primary care settings as well as new roles for clinical pharmacists. PMID- 11918504 TI - Hyperlipidemia in HIV-positive patients receiving antiretrovirals. AB - OBJECTIVE: To assess the frequency of lipid abnormalities and treatment outcomes for hyperlipidemia in HIV-positive patients receiving antiretroviral (ARV) therapy as outpatients at a Veterans Affairs HIV clinic. METHODS: All patients monitored for at least 3 months were reviewed. Data collected included age, most recent CD4+ cell count and viral load, ARV history, and all fasting cholesterol, triglyceride, high-density lipoprotein (HDL), and low-density lipoprotein (LDL) values. The ARV therapy at the time of lipid readings was classified as including protease inhibitors (PI+) or not including them (PI-). Lipid values were compared with goals per national guidelines and risk factors. RESULTS: Male patients (n = 101) providing 210 lipid profiles were evaluated (median 2 per patient). Median age was 51 years. Fourteen patients were diabetic, 31 were hypertensive, and 6 patients had documented coronary disease. Mean cholesterol, triglycerides, and LDL values were significantly higher in PI+ (n = 50) compared with those of PI- patients (n = 51; p < 0.05). HDL values were not different between groups. Significantly more PI+ patients had lipid concentrations above recommended goals compared with PI- patients (17 vs. 7; p = 0.04). Six patients achieved lipid goals after following a low-fat diet (4 PI+). Fifteen subjects (11 PI+) were being treated with medications. Ten patients (67%) reached lipid goals, 2 had not reached goals (13%), and 3 (20%) were undergoing medication titration. CONCLUSIONS: Our HIV-infected patients had significantly higher cholesterol, triglyceride, and LDL values when PI+. In contrast to other reports, the majority of patients treated for lipid abnormalities achieved treatment goals. PMID- 11918505 TI - A decade of antimicrobial susceptibilities at the University of Kentucky Hospital. AB - OBJECTIVE: To determine the antimicrobial susceptibility rates for key antimicrobial agents and selective bacterial pathogens in the decade of the 1990s. METHODS: Data from 1990 to 2000 from the University of Kentucky Clinical Microbiology Laboratory were analyzed by linear regression analysis to identify agents and pathogens that show a decline in susceptibility. For selected pathogens and antimicrobial agents, predictions were made for further declines in susceptibility for 2005 and 2010. RESULTS: Significant declines in susceptibility to selected antimicrobial agents were found for Pseudomonas aeruginosa, Klebsiella pneumoniae, Enterobacter cloacae, Escherichia coli, Staphylococcus aureus, Staphylococcus epidermidis, and Streptococcus pneumoniae. Further declines were predicted for 2005 and 2010. CONCLUSIONS: Examination of susceptibility rates over time in a university hospital medical center provides useful data for future planning. In our institution, antimicrobial susceptibility rates have significantly declined during the 1990s for certain antimicrobial agents and bacterial pathogens. We are attempting to change our antimicrobial use patterns through formulary manipulation and clinician education, which may retard or prevent such declines in the future. PMID- 11918506 TI - The economic burden of Barrett's esophagus in a Medicaid population. AB - OBJECTIVE: To estimate the overall healthcare expenditures of patients with Barrett's esophagus in the West Virginia Medicaid population. METHODS: West Virginia Medicaid-paid claims data for the period January 1, 1995, to December 31, 1999, were used for the study. The population included all individuals eligible for West Virginia Medicaid during the study period except for Medicare eligible- and Medicaid managed-care recipients. A prevalence-based approach was used to determine the cost of illness for Barrett's esophagus. RESULTS: The total cost of illness for Barrett's esophagus more than tripled, from $182399 in 1995 to $623864 in 1999, with approximately a 4(1/2)-fold increase in medical and more than a threefold increase in pharmacy costs. The average cost of treating Barrett's esophagus was found to be approximately $1207 per patient in 1999. Overall, pharmacy costs accounted for >66% of the total costs. Controlling for age, gender, and number of comorbidities, patients with Barrett's esophagus incur 21.2% higher overall costs than patients with gastroesophageal reflux disease and 62.4% higher overall costs than the general Medicaid population. CONCLUSIONS: The increasing prevalence of and resource utilization for Barrett's esophagus provide a framework for further analysis and implementation of policies aimed at appropriate allocation of resources for the state's Medicaid program. PMID- 11918507 TI - Headache education in colleges of pharmacy. AB - OBJECTIVE: To assess primary headache education in colleges of pharmacy during the 2000-2001 academic year. DESIGN: A survey seeking to quantify students' primary headache education was mailed to all 82 American Association of Colleges of Pharmacy member schools. RESULTS: A usable response was obtained from 65 of 74 answering schools. Per professional year, the average pharmacy student receives 1 headache core course contact hour and no headache elective course contact hours. Two schools offer clerkships devoted exclusively to headaches. Six schools offer clerkships in which a student could expect to focus on headache therapy at least 25% of the time. Seven schools plan to alter their current curriculum to include more headache education, all via additional lectures. CONCLUSIONS: Few opportunities exist for students to learn about primary headaches in colleges of pharmacy. Given the high prevalence and poor medical management of primary headache disorders along with the commonality of pharmacist-headache patient interactions occurring in practice, pharmacy schools should evaluate and alter their curriculum to include more primary headache education. PMID- 11918508 TI - Effect of stress on international normalized ratio during warfarin therapy. AB - OBJECTIVE: To discuss the effect of stress on the international normalized ratio (INR) when patients are taking warfarin. CASE SUMMARY: Two patients at a pharmacist-managed anticoagulation clinic who were stable with anticoagulation developed elevated INR values after a stressful event occurred. All other factors known to elevate the INR were unchanged; furthermore, the INR values returned to the prior level of control after resolution of the stressful events. DISCUSSION: Management of anticoagulation with warfarin requires the knowledge of factors that may alter an INR. Many of these factors, such as dietary changes, illnesses, drug interactions, patient compliance, and physical activity, have been described. In spite of this understanding, many patients continue to experience variability in their INR values, suggesting there are other factors that can alter the INR that have not been fully described. The cases presented here demonstrate that stressful events, physical or psychological, can elevate the INR. The mechanism for this occurrence is unknown, but may be related to decreased metabolism of warfarin during stress. CONCLUSIONS: When an unexplained INR value exists, a stressor should be evaluated as a potential cause. PMID- 11918509 TI - Severe interaction between ritonavir and acenocoumarol. AB - OBJECTIVE: To report a clinically severe interaction between ritonavir (RTV) and acenocoumarol resulting in a decrease in the anticoagulant effect severe enough to eventually preclude RTV administration. CASE SUMMARY: An asymptomatic, HIV infected, 46-year-old man with mitraortic prosthetic valves receiving acenocoumarol therapy started stavudine, lamivudine, and RTV 600 mg twice daily. His international normalized ratio (INR) decreased dramatically (the opposite of what should be expected). Although the acenocoumarol dose was progressively increased to 3 times the original dose, it was impossible to achieve the previous INR and RTV was withdrawn. DISCUSSION: RTV is an inducer of the hepatic isoenzymes CYP1A2, CYP1A4, and CYP2C9/19 and leads to extensive metabolism of acenocoumarol that cannot be balanced by dose increases. This effect is the opposite of what was expected to occur, considering that RTV is also a strong inhibitor of most hepatic isoenzymes. CONCLUSIONS: RTV severely decreases the anticoagulant effect of acenocoumarol. It must be added to the list of drugs that affect the action of oral anticoagulants, and it probably should be avoided in patients receiving acenocoumarol. PMID- 11918510 TI - Blood pressure elevation in a patient treated with salsalate. AB - OBJECTIVE: To report a case of increased blood pressure associated with the use of salsalate in an elderly patient with no prior history of hypertension. CASE SUMMARY: A 78-year-old white man with no prior history of hypertension initiated salsalate therapy for low-back pain. Over the 15 months prior to the initiation of salsalate, his blood pressure averaged 127 +/- 7 mm Hg systolic and 84 +/- 6 mm Hg diastolic (mean +/- SD). After initiation of salsalate, he experienced significant elevations in blood pressure, which led to a preliminary diagnosis of hypertension. Blood pressure after initiation of salsalate averaged 150 +/- 13 mm Hg systolic and 95 +/- 5 mm Hg diastolic. No changes in medications or medication doses (with the exception of warfarin) occurred in the 18 months prior to or during salsalate therapy. His weight remained stable. A detailed review of his medical records and history revealed no other causes for these elevations in blood pressure. Salsalate therapy was discontinued and his blood pressure returned to normotensive levels (119 +/- 2 mm Hg systolic and 81 +/- 2 mm Hg diastolic). DISCUSSION: Nonsteroidal antiinflammatory drug (NSAID)-induced elevations in blood pressure have been well documented in patients receiving antihypertensive medications. Due to its relative weak inhibition of cyclooxygenase and lack of published literature in hypertensive patients, salsalate is considered to have little or no effect on blood pressure. Our report documents a possible case of salsalate-induced hypertension in a previously normotensive elderly man. Observational studies suggest that NSAID use may increase the risk of developing hypertension in older patients. CONCLUSIONS: Clinicians should be aware of the possible effects of NSAIDs on blood pressure. Blood pressure monitoring following the initiation of salsalate may be warranted, particularly in older patients. PMID- 11918511 TI - Reversible thrombocytopenia associated with eptifibatide. AB - OBJECTIVE: To report a patient who experienced severe, reversible thrombocytopenia after receiving eptifibatide for acute coronary syndrome. CASE SUMMARY: A 61-year-old white man with a past medical history of coronary artery disease, peripheral vascular disease, insulin-dependent diabetes, hypertension, and dyslipidemia was treated with eptifibatide as an adjunct to standard antiischemic therapy for acute coronary syndrome. On administration of eptifibatide, the patient experienced chills accompanied by sharp pains that radiated throughout his body. The infusion was immediately stopped and the symptoms resolved. A platelet count obtained 11 hours later showed a decrease from a baseline of 230 to 3 x 10(3)/mm(3) (normal 160-360). Multiple platelet analyses confirmed profound thrombocytopenia. Over the course of the hospitalization, the patient's platelet count returned to normal; he experienced no other adverse hematologic sequelae. DISCUSSION: All of the glycoprotein IIb/IIIa receptor antagonists are associated with thrombocytopenia. However, the risk of thrombocytopenia has been observed to be less with eptifibatide. Because of the temporal relationship between eptifibatide administration and the patient's symptoms and laboratory analysis, we believe that the thrombocytopenia was the result of an adverse reaction. CONCLUSIONS: The temporal relationship to administration and the resolution of the adverse reaction on discontinuation of the drug support the likelihood that the severe, reversible thrombocytopenia was associated with eptifibatide. PMID- 11918513 TI - Treatment of resistant depression by adding noradrenergic agents to lithium augmentation of SSRIs. AB - OBJECTIVE: To examine the efficacy of second-line augmentation with noradrenergic antidepressants (NAs) in depressed patients who partially responded to lithium augmentation of selective serotonin-reuptake inhibitors (SSRIs). CASE SUMMARY: Six patients with major depression or double depression (major depression and dysthymia) who were partially responsive to lithium and SSRI treatment were given either bupropion or desipramine, in an open clinical manner. Improvement was determined and rated by a psychiatrist based on clinical judgment guided by the Clinical Global Impression (CGI) improvement scale and by the Global Assessment of Functioning (GAF) as described in Axis V in the Diagnostic and Statistical Manual of Mental Disorders, Fourth Edition. RESULTS: Among the 6 depressed patients with partial remission (much improved in symptoms and moderate functional improvement: CGI score 2, GAF score 51-60) while taking the SSRI and lithium combination, 2 showed complete remission (very much improved in symptoms and good functioning: CGI 1, GAF 80-100) and 3 achieved near-complete remission (very much improved in symptoms and significant functional recovery: CGI 1, GAF 61-80) when given either bupropion or desipramine. One patient did not show any additional clinical or functional improvement. Second-line augmentation with bupropion was better tolerated than desipramine. CONCLUSIONS: This clinical observation suggests that second-line augmentation with NAs may be a viable option to optimize recovery in depressed patients with a partial response to lithium augmentation of SSRIs. PMID- 11918512 TI - Cross-sensitivity between paroxetine and sertraline. AB - OBJECTIVE: To report a case of possible cross-sensitivity between selective serotonin-reuptake inhibitors (SSRIs). CASE SUMMARY: A 20-year-old Southeast Asian man developed a maculopapular rash soon after starting paroxetine. Following resolution of this rash, another skin reaction with the same distribution and appearance occurred after sertraline therapy was started. DISCUSSION: Cross-reactivity between drugs with similar structures has been reported; however, cross-reactivity among SSRI antidepressants is unexpected given their differences in chemical structure. CONCLUSIONS: The possibility of cross-reactivity between SSRI antidepressants should be considered by clinicians who wish to switch from one SSRI to another due to a dermatologic reaction. PMID- 11918514 TI - Severe serotonin syndrome induced by mirtazapine monotherapy. AB - OBJECTIVE: To document a case of serotonin syndrome (SS) associated with mirtazapine monotherapy, review the previously reported cases of SS associated with this tetracyclic antidepressant, and discuss the possible pathogenic mechanisms leading to this serious adverse drug reaction. CASE SUMMARY: A 75-year old man developed agitation, confusion, incoordination, and gait disturbance because of progressive rigidity. Mirtazapine had been started 8 days earlier to control major depression. Physical examination revealed diaphoresis, low-grade fever, hypertension, tachycardia, bilateral cogwheel rigidity, hyperreflexia, tremor, and myoclonus, symptoms and signs that are consistent with severe SS. DISCUSSION: A review of the cases of SS with implication of mirtazapine as the cause was performed. The possible pathogenic mechanisms leading to this adverse reaction in this patient are also discussed, and pathophysiologic hypotheses are formulated. CONCLUSIONS: Although mirtazapine offers clinicians a combination of strong efficacy and good safety, we suggest bearing SS in mind when prescribing this drug, especially in frail, elderly patients with underlying chronic conditions. In these patients, it might be more adequate to start mirtazapine therapy at a lower dose (<15 mg/d). PMID- 11918515 TI - Neonatal cholestatic hepatitis from carbamazepine exposure during pregnancy and breast feeding. AB - OBJECTIVE: To report a case of transient cholestatic hepatitis occurring in an infant between the third and seventh weeks of life, most likely due to carbamazepine exposure during pregnancy and breast feeding. CASE SUMMARY: A boy, born to an epileptic mother who had been treated with carbamazepine monotherapy throughout pregnancy and breast feeding, experienced asphyxia at birth with transient hepatic dysfunction in the first week of life. After full recovery from asphyxia, he experienced a second period of liver dysfunction, presenting as cholestatic hepatitis that lasted approximately 5 weeks. Infectious and metabolic etiologies as well as extrahepatic biliary atresia were excluded. DISCUSSION: Carbamazepine is known to induce hepatic damage in children and adults. As the drug crosses the placenta and is excreted into breast milk, infants of mothers taking carbamazepine might also develop liver dysfunction. In addition to the present case, there are 2 well-documented case reports of cholestasis in association with transplacental and transmammary carbamazepine exposure. CONCLUSIONS: Carbamazepine-induced hepatitis may occur in association with prenatal exposure and breast feeding. This may expose infants to unnecessary diagnostic procedures, and should therefore be mentioned in the company's product information. PMID- 11918516 TI - New and emerging therapies for sepsis. AB - OBJECTIVE: To review the recent advances related to the pathophysiology of sepsis and the rationale for recombinant human-activated protein C (drotrecogin alfa) and other antisepsis agents currently in Phase III trials. DATA SOURCES: A MEDLINE (1990-December 2001) search was performed to identify pertinent literature on the pathophysiology of sepsis and treatment strategies. The search was supplemented with AdisInsight (Adis International) using the search terms sepsis, severe sepsis, or septic shock combined with agents in Phase II or higher clinical development. Abstracts presented at infectious diseases and critical care meetings were also reviewed. STUDY SELECTION AND DATA EXTRACTION: Clinical efficacy studies were selected for drotrecogin alfa and other Phase III investigational agents. DATA SYNTHESIS: Our current understanding of the pathophysiology of sepsis underscores the contribution of increased coagulation and diminished fibrinolytic activity working in conjunction with an excessive and dysregulated inflammatory response. The loss of homeostatic balance among these systems results in a systemic inflammatory response with generalized coagulopathy, microvascular thrombosis, and, ultimately, acute organ failure and death. As a result of these advances, several compounds are now in various phases of development. A recombinant human form of endogenous activated protein C (drotrecogin alfa) was recently approved by the Food and Drug Administration for severe sepsis in adults who have a high risk of death. It possesses anticoagulant, profibrinolytic, and antiinflammatory properties. Other compounds currently in Phase III trials include tissue-factor pathway inhibitor, tumor necrosis factor antibody fragment, platelet-activating factor acetylhydrolase, antithrombin III, and pyridoxylated hemoglobin polyoxyethylene. CONCLUSIONS: With the recent approval of drotrecogin alfa, there is renewed optimism that we can effectively reduce sepsis-associated mortality. PMID- 11918517 TI - Esomeprazole for acid peptic disorders. AB - OBJECTIVE: To review the pharmacology, pharmacokinetics, clinical efficacy, and safety of esomeprazole, a new proton-pump inhibitor (PPI). DATA SOURCES: A MEDLINE search (1966-March 2001) was conducted for relevant literature using the terms esomeprazole, Nexium, and H199/18. Abstracts from the XXXII Nordic Meeting of Gastroenterology and journal articles provided by AstraZeneca were reviewed. STUDY SELECTION AND DATA EXTRACTION: All available studies on the pharmacology, pharmacokinetics, clinical efficacy, and safety of esomeprazole were reviewed. DATA SYNTHESIS: Esomeprazole is a new PPI and is the S-isomer of racemic omeprazole. Esomeprazole has demonstrated acid control comparable to that of the other PPIs currently available. Esomeprazole undergoes less hepatic metabolism compared with omeprazole, and thus may result in less interpatient variability among slow and fast metabolizers of CYP2C19. The oral bioavailability of esomeprazole is approximately 89% with a dose of 40 mg, and the half-life is approximately 1.5 hours. Esomeprazole is effective in the healing of erosive esophagitis, with a rate of healing at week 8 of 93.7% (p < 0.001). In maintenance therapy of healed erosive esophagitis, esomeprazole maintained healing rates >90% (6-mo trial). Esomeprazole is comparable with omeprazole (both in combination with appropriate antibiotics) in the eradication of Helicobacter pylori, with eradication rates of 89.7% and 87.8%, respectively. The drug is well tolerated. The few adverse effects associated with esomeprazole are diarrhea, headache, nausea, abdominal pain, respiratory infection, and sinusitis. CONCLUSIONS: Esomeprazole is a safe and effective PPI. It is effective in the treatment of peptic ulcer disease and gastroesophageal reflux disease. PMID- 11918518 TI - Risedronate for the prevention of fractures in postmenopausal osteoporosis. AB - OBJECTIVE: To evaluate current scientific literature regarding the efficacy of risedronate in the prevention of vertebral and nonvertebral fractures in women with postmenopausal osteoporosis. DATA SOURCES: Primary research articles were identified by MEDLINE search (1966-May 2001) and through secondary sources. Key search terms were risedronate, postmenopausal osteoporosis, and fractures. DATA SYNTHESIS: Osteoporosis results in a reduction of bone mineral density, increased bone fragility, and increased risk of fractures. The goal of osteoporosis therapy is not only to increase bone mass, but also to reduce the rate of fractures. Risedronate is the newest bisphosphonate to be approved for the prevention and treatment of osteoporosis. An evaluation of clinical trials using risedronate in the treatment of postmenopausal osteoporosis was performed to determine its efficacy at decreasing fracture rates. CONCLUSIONS: Risedronate is an effective and safe option for the treatment of postmenopausal osteoporosis. Risedronate significantly decreases the risk of vertebral and nonvertebral fractures in women who have had > or =1 fractures in the past. More studies are warranted to evaluate the efficacy of risedronate in women without preexisting vertebral fractures. PMID- 11918519 TI - Potential use of HMG-CoA reductase inhibitors for osteoporosis. AB - OBJECTIVE: To review the current clinical data describing the effects of hydroxymethylglutaryl coenzyme A (HMG-CoA) reductase inhibitors in bone formation and reduction of fracture incidence and their potential use for osteoporosis. DATA SOURCES: English-language articles and abstracts were identified from a MEDLINE database search (1966-May 2001) that used the key words osteoporosis, HMG CoA reductase inhibitors, and treatment. STUDY SELECTION AND DATA EXTRACTION: All the articles identified from the data sources were evaluated, and all information deemed relevant was included in this review. DATA SYNTHESIS: In the past, there has been considerable emphasis placed in the detection, prevention, and treatment of osteoporosis. New drug therapies have been introduced in the market aimed at decreasing bone loss and increasing bone formation, with the ultimate goal of decreasing fractures. Currently, there are a variety of agents available for the treatment of osteoporosis. A limited number of case-control studies have suggested that HMG-CoA reductase inhibitors (statins) may have the potential to reduce the risk of fractures by increasing bone formation, although other studies have failed to show a benefit in fracture reduction. The potential benefit of this therapy is still undetermined because of a lack of randomized, controlled, clinical trials and conflicting data. CONCLUSIONS: At this time, there are limited data on the role of statins in the treatment of osteoporosis. Practitioners must be cautious when using these agents over other proven conventional therapies. Randomized, controlled, clinical trials are needed to accurately determine the role of these agents in the treatment of osteoporosis. PMID- 11918520 TI - Pathophysiology and first-line treatment of osteoarthritis. AB - OBJECTIVE: To review the pathophysiology of osteoarthritis (OA) and the various treatment modalities, focusing specifically on acetaminophen (APAP), nonsteroidal antiinflammatory drugs (NSAIDs), and cyclooxygenase-2 (COX-2) inhibitors as the primary treatment options. DATA SOURCES: Primary literature and tertiary references were identified by a MEDLINE search (1966-March 2001) and through other secondary sources. STUDY SELECTION AND DATA EXTRACTION: After evaluating the articles and references identified from the data sources, all the information that was judged relevant by the reviewers was included in the review article. DATA SYNTHESIS: OA is the most common joint disorder worldwide. Current research suggests that factors such as inflammation and changes in subchondral bone may play a larger role in the pathophysiology than previously thought. With this research and the development of COX-2 inhibitors, selecting the medication of choice for OA has become difficult. CONCLUSIONS: More research needs to be done before the pathophysiology of OA can be clearly determined. In the meantime, treatment should be based on clinical data and patient response. Studies have shown that APAP and NSAIDs have comparable efficacy, as do traditional NSAIDs and COX-2 inhibitors. APAP is associated with fewer toxicities than are the traditional NSAIDs. Due to their mechanism of action, the new COX-2 inhibitors should result in fewer adverse effects compared with traditional NSAIDs, but evidence from clinical trials has not been conclusive. Therefore, APAP should still be considered the drug of choice for OA. PMID- 11918521 TI - Lidocaine-prilocaine cream versus tetracaine gel for procedural pain in children. AB - OBJECTIVE: To determine the relative efficacy of lidocaine-prilocaine and tetracaine for procedural pain in children. METHODS: Systematic review was performed (MEDLINE1990-June 2001) of all studies comparing the efficacy of these 2 modalities of pain relief in children undergoing painful cutaneous procedures. Search terms included lidocaine, prilocaine, tetracaine, and anesthesia. RESULTS: Eight studies compared lidocaine-prilocaine with tetracaine in children for 4 different procedures: intravenous cannulation, venipuncture, Port-a-Cath puncture, and laser therapy. When used as labeled (60 min for lidocaine prilocaine, 30 min for tetracaine), the 2 modalities provided similar analgesic efficacy. When both anesthetics were applied for a similar duration of time (40 min, 60 min, 2 h), tetracaine provided superior anesthesia. Tetracaine was commonly associated with erythema, and lidocaine-prilocaine was associated with blanching of the skin. CONCLUSIONS: Lidocaine-prilocaine and tetracaine appear to be comparable for procedural pain relief when used as recommended. Tetracaine is more efficacious than lidocaine-prilocaine when both anesthetics are applied for the same amount of time. PMID- 11918522 TI - Stereoselective pharmacokinetics and pharmacodynamics of anti-asthma agents. AB - OBJECTIVE: To review the previously published studies on pharmacokinetics and pharmacodynamics of chiral drugs used in the treatment of asthma. DATA SOURCES: Primary and review articles were identified with a MEDLINE search (1980-May 2001) and through secondary sources. STUDY SELECTION AND DATA EXTRACTION: All English language studies and reviews obtained from the MEDLINE search pertaining to stereoselective pharmacokinetics and pharmacodynamics of chiral anti-asthma drugs were assessed. DATA SYNTHESIS: Several anti-asthma drugs (e.g., beta(2) adrenergic agonists, leukotriene modifiers) are chiral and marketed as racemates, which consist of equal proportions of 2 enantiomers. Significant stereoselectivity has also been reported in pharmacodynamics and pharmacokinetics of the beta(2)-agonists. The enantiomers of beta(2)-agonists in the R configuration are primarily responsible for the bronchodilating effects of the racemate. The plasma concentrations of the enantiomers of anti-asthma drugs may differ as a reflection of stereoselectivity in clearance, volume of distribution, and route of administration. CONCLUSIONS: Stereoselectivity in the pharmacokinetics of anti-asthma drugs may complicate the relationship between dose and/or plasma concentration of racemic drug versus effect relationship. An appreciation of the stereoselective pharmacokinetics and pharmacodynamics of chiral anti-asthma drugs may optimize the use of these agents in asthmatic patients. PMID- 11918523 TI - Low-dose ritonavir for protease inhibitor pharmacokinetic enhancement. AB - OBJECTIVE: To examine the use of low-dose ritonavir as a pharmacokinetic enhancer for HIV protease inhibitors. DATA SOURCES: Primary articles, review articles, and conference abstracts identified by MEDLINE search (1995-May 2001) and secondary sources. DATA SYNTHESIS: Low-dose ritonavir (100-200 mg) is increasingly being combined with HIV protease inhibitors to improve their effectiveness and allow less frequent dosing. An evaluation of the clinical evidence supporting this practice was conducted. CONCLUSIONS: Limited outcome data exist for low-dose ritonavir-based regimens in general. Although preliminary data appear promising, more clinical evidence is needed to determine the optimal dosing, long-term safety, and relative effectiveness of this approach. The role of these regimens in early therapy remains to be defined. PMID- 11918524 TI - Serotonergic agents in the treatment of fibromyalgia syndrome. AB - OBJECTIVE: To evaluate literature that discusses the treatment of fibromyalgia syndrome (FMS) with agents that involve the neurotransmitter serotonin. DATA SOURCES: Biomedical literature accessed through MEDLINE (1966-August 2001) and International Pharmaceutical Abstracts. DATA SYNTHESIS: The cause and pathophysiology of FMS remain elusive, although abnormalities in the serotonin pathway have been implicated. Several serotonergic agents have been studied for use in FMS. Trials and case reports focusing on the use of newer agents: the selective serotonin reuptake inhibitors, venlafaxine and tramadol, were reviewed. CONCLUSIONS: Current research suggests that the serotonergic agents may reduce at least some of the symptoms of FMS. However, medications that act on multiple neurotransmitters may prove to be more effective in symptom management. Additional long-term studies are required in order to validate these results. PMID- 11918525 TI - Fluoxetine in the treatment of premenstrual dysphoric disorder. AB - OBJECTIVE: To examine the role of fluoxetine in the treatment of premenstrual dysphoric disorder (PMDD). DATA SOURCES: Search strategy included MEDLINE (1966 February 2002), Embase (1988-February 2002), HealthStar (1975-December 2000), Current Contents (1996-November 2001), and Copernic (November 2001). Search terms included fluoxetine, premenstrual dysphoric disorder, PMDD, late luteal-phase dysphoric disorder, and severe premenstrual syndrome. STUDY SELECTION: English language human studies were selected and evaluated based on quality of evidence. DATA SYNTHESIS: Eight prospective trials (3 double-blind, placebo-controlled, crossover; 3 double-blind, randomized, controlled; 2 open-label), 1 case series, and 1 meta-analysis were identified. Although 6 of the studies involved small sample sizes (n < 50), all found fluoxetine to be effective in the treatment of PMDD. CONCLUSIONS: Despite limited data, fluoxetine 20 mg/d appears to be effective in the treatment of PMDD. However, adverse effects, particularly headaches and sexual dysfunction, are possible. Given the long half-life of fluoxetine and the short duration of PMDD symptoms per cycle, larger, well designed clinical trials evaluating intermittent dosing for only 1 week or a few doses need to be performed. PMID- 11918526 TI - Mantle cell lymphoma in leukaemic phase. PMID- 11918527 TI - Cyclophosphamide-induced nail pigmentation. PMID- 11918528 TI - Historical aspects of paroxysmal nocturnal haemoglobinuria: 'defining the disease'. PMID- 11918529 TI - Immunomodulatory gene therapy for haematological malignancies. PMID- 11918530 TI - Results of treatment of children with refractory anaemia with excess blasts (RAEB) and RAEB in transformation (RAEBt) in Great Britain 1990-99. AB - Between 1990 and 1999, 36 children with refractory anaemia with excess blasts (RAEB) and RAEB in transformation (RAEBt), not associated with Down's syndrome, were diagnosed in Britain. A total of 31 children received intensive chemotherapy, six of whom proceeded to a bone marrow allograft in first remission, whereas two received an autograft. Of the 23 given chemotherapy only, four died of toxicity, 10 relapsed and nine are alive in first remission. Out of the 10 who relapsed, four are alive and disease-free following an allograft. Out of the 6 children given an allograft in first remission, two died of disease and four are alive in first remission. Both children given an autograft died of disease. Two children received an allograft without prior chemotherapy but died of toxicity. Three children received supportive care only, and one child survived. The overall survival was 51% at 5 years, and was superior in children with RAEBt (63%) compared with RAEB (28%, P = 0.03). Cytogenetics were available in 35 cases. Monosomy 7 was the most common abnormality (33% of cases). Survival in children with monosomy 7 was 22% at 5 years compared with 66% for the other patients (P = 0.05). Allowing for cytogenetics, outcomes of therapy appear similar to those for de novo acute myeloid leukaemia (AML), and it is appropriate for children with RAEB/RAEBt to be registered in AML trials. PMID- 11918531 TI - Effect of fluconazole prophylaxis on fungal blood cultures: an autopsy-based study involving 720 patients with haematological malignancy. AB - To investigate the utility of blood culture of invasive fungal infections in patients with haematological malignancies, an autopsy survey was conducted in 720 patients who were treated between 1980 and 1999. We identified 252 patients with invasive mycosis. These included Candida (n = 94), Aspergillus (n = 91), Zygomycetes (n = 34), Cryptococcus (n = 7), Trichosporon (n = 11), Fusarium (n = 1), and unknown fungi (n = 20). Of the 94 patients with invasive candidiasis, 20 had positive blood cultures. Of the 11 patients with invasive trichosporonosis, seven had positive blood cultures. The sensitivities of blood cultures were 1.1%, 0% and 14% for detecting invasive aspergillosis, zygomycosis and cryptococcosis respectively. Multiple regression analysis showed a significant correlation between results of Candida blood cultures and some variables, including prophylactic use of absorbable antifungals (P = 0.0181) and infection by Candida albicans (P = 0.0086). The sensitivity of blood cultures decreased when patients received antifungal chemoprophylaxis. Unless these agents are inactivated in culture bottles, conventional blood cultures might produce false-negative results. PMID- 11918532 TI - Serum erythropoietin values in erythrocytoses and in primary thrombocythaemia. AB - Serum erythropoietin (Epo) values were estimated in samples from 125 patients with erythrocytosis to examine the specificity and sensitivity of reduced and raised values in the diagnosis of polycythaemia vera (PV) and secondary erythrocytosis (SE) respectively. Additionally, Epo values were estimated in samples from 49 patients with primary thrombocythaemia (PT) to determine whether Epo values were altered. We found high specificity (92%) and moderate sensitivity (64%) of low serum Epo values (below the reference range) in the diagnosis of PV, and also poor sensitivity (47%) of raised Epo values in the diagnosis of SE. Raised Epo values were not observed in PV patients with Hb > 14.0 g/dl and were only observed in one PV patient with a relatively low Hb recovering from a gastro intestinal haemorrhage. Raised Epo values occurred in some patients with apparent erythrocytosis (AE) and idiopathic erythrocytosis (IE), mainly at normal (rather than raised) Hb values (< 16 g/dl). Low Epo values occurred in a few AE, IE and SE patients at higher Hb values (> 16 g/dl). Low Epo values were less specific for PV when the Hb was raised, while raised Epo values were less specific for SE when the Hb was not raised. Approximately one third of patients with PT had a low (below the reference range) Epo value, this being associated with a high normal Hb (> 14 g/dl, P < 0.001) and showing a trend towards association with absence of treatment. The high normal Hb values were in turn associated with an increased incidence of thrombotic events (P < 0.05). These findings could influence the future investigation and management of PT patients. PMID- 11918533 TI - Prognostic value of soluble interleukin 2 receptor levels in Langerhans cell histiocytosis. AB - We investigated the prognostic significance of soluble interleukin 2 receptor (sIL-2r) levels in the pre- and post-treatment serum of paediatric patients with Langerhans cell histiocytosis (LCH). Serum levels of sIL-2r from 32 LCH patients and 14 healthy controls were determined using enzyme-linked immunosorbent assay. The LCH patients were classified, evaluated and treated according to the Histiocyte Society's protocols. The following clinical stages were considered: single-system disease (A) divided into single-site (A1; n=4), multiple-site (A2; n=9), and multisystem disease (B) without organ dysfunction (B1; n=5) and with organ dysfunction (B2; n=14). Pretreatment concentrations of sIL-2r were markedly increased at diagnosis in LCH patients compared with controls [in pg/ml, median (range) 9200 (1124-40000) versus 610 (343-800)], P < 0.0001. Levels differed significantly between stages A [3250 (1124-11000)] and B [22750 (3400-40000)], P < 0.05, and between substages A2 and B2, P < 0.05. There was a significant correlation between clinical stages and sIL-2r serum levels, r=0.7996 (P < 0.0001). Patients with > or = 17500 pg/ml of sIL-2r had a 30-month survival of 0.417 (SEM: 0.142) compared with those with levels < 17500 pg/ml, who presented a 30-month survival of 0.848 (SEM: 0.100) (log-rank, P < 0.0001). In multivariate analysis, sIL-2r levels > or = 17500 pg/ml were found to have greater predictive strength than other well-known prognostic factors. PMID- 11918534 TI - Co-expression of CD30 ligand and interleukin 4 (IL-4) receptors by acute myeloid leukaemia blasts is associated with the expansion of IL-4-producing CD30+ normal T cells. AB - CD30 ligand (CD30L), but not its cognate receptor CD30, is frequently expressed on acute myeloid leukaemia (AML) blasts. In the present study, we found that leukaemic blasts presenting surface CD30L displayed a characteristic cytokine receptor pattern that makes them ideal targets for those cytokines usually produced by Th2-type cell subsets. In particular, even though a broad distribution of Th2 cytokine receptors by AML blasts was shown, we demonstrated the almost exclusive expression of interleukin 4 (IL-4) receptor (R), in the absence of its cognate cytokine, by CD30L+ AML. Furthermore, a number of Th2 associated markers, including CD30, IL-4 and GATA-3, were expressed by residual T cells derived from CD30L+ AML but not from CD30L- AML, in which the presence of the Th1-associated marker LAG-3 was documented in some cases. The production of IL-4 in the absence of interferon gamma (IFN-gamma) was also detected in CD3+/CD30+ T cells from CD30L+ AML. These results, along with the shift toward IL 4-producing specific T-cell clones observed in CD30L+ AML samples by enzyme linked Immunospot (ELISpot) assay, were consistent with the hypothesis of a Th2 polarization taking place in T cells from CD30L+ AML. The notion that IL-4 was able to enhance in vitro proliferation of CD30L+/IL-4R+ purified leukaemic blasts suggests that the selective interaction of IL-4-producing CD30+ T cells with CD30L+ leukaemic progenitors may have a role in the progression of this particular AML subset. PMID- 11918535 TI - Magnetic resonance imaging of femoral marrow predicts outcome in adult patients with acute myeloid leukaemia in complete remission. AB - Accurate assessment of residual disease is important for the prediction of outcome in patients with acute leukaemia in complete remission (CR). To investigate whether abnormalities on magnetic resonance (MR) images of femoral marrow in adult patients with acute myeloid leukaemia (AML) in CR can predict outcome, 28 newly diagnosed patients with AML underwent MR imaging when bone marrow aspiration or biopsy was performed to verify the state of CR after induction therapy. MR abnormalities on short TI (inversion time) inversion recovery (STIR) techniques persisted in all four patients who did not achieve CR. In 13 CR patients abnormalities on STIR images resolved, to result in normal appearance at the time CR was achieved. All 13 patients remained in CR for 3-104 months (median, 73 months). In the other 11 CR patients, STIR abnormalities persisted at the time CR was achieved. Seven of them relapsed between 1 and 28 months (median, 3 months) after MR evaluation. Disease-free survival of patients with persistent abnormal STIR images was significantly shorter than that of patients with normal STIR images (P < 0.01). MR imaging of femoral marrow may predict outcome in adult patients with AML in CR. PMID- 11918536 TI - Differentiation of promyelocytic leukaemia: alterations in Fas (CD95/Apo-1) and Fas ligand (CD178) expression. AB - The survival of leukaemic blasts contributes to the pathological mechanism of acute promyelocytic leukaemia (APL). While treatment of APL using retinoic acid (RA) is a model of differentiation therapy, little is known about possible effects of this treatment on the Fas/FasL system. Investigation of APL cells from patients undergoing differentiation therapy with RA and of promyelocytic HL-60 and monoblastic U-937 cells cultured with RA revealed a reduction of surface expression of both Fas and its ligand. Accordingly, the sensitivity of the cells to anti-Fas-induced apoptosis decreased proportionally and the reduced expression of FasL resulted in a decreased ability of the leukaemic cells to induce apoptosis in T cells. Our findings demonstrate that there are significant changes in Fas and FasL expression during RA treatment of APL, which probably have consequences for the interaction between host immune and leukaemia cells, and thus may be involved in the beneficial effects of differentiation therapy. PMID- 11918537 TI - RANK (receptor activator of nuclear factor-kappaB) and RANKL expression in multiple myeloma. AB - The new members of the tumour necrosis factor (TNF) receptor-ligand family, receptor activator of nuclear factor-kappaB ligand (RANKL) and its receptor RANK, play a crucial role in osteoclast differentiation and activation. An increased expression of RANKL and/or RANK may be involved in the excessive bone resorption observed in multiple myeloma (MM). We used immunohistochemistry to study RANK and RANKL expression in bone marrow (BM) biopsies obtained at diagnosis in 15 MM patients, six patients with monoclonal gammopathy of undetermined significance (MGUS) and 10 normal BM biopsies. Plasma cells were not labelled with anti-RANKL or anti-RANK antibodies. In all biopsies, RANKL was expressed in endosteal bone surface, around vessels and in cells characterized by cytoplasmic expansions. These last cells did not express CD45 and were vimentin positive, corresponding to bone marrow stromal cells. Numerous stromal cells expressed RANKL in MM and MGUS specimens, with a greater expression in MM than in MGUS. Very few cells were stained with anti-RANKL in normal BM specimens. With the anti-RANK antibody, small mononuclear cells in the bone microenvironment were positive and were identified as erythroblast cells. In conclusion, we showed that RANKL was expressed in reticular stromal cells, with a greater intensity in myeloma specimens. These results suggest that RANKL overexpressed by bone marrow stromal cells may contribute to the high rate of bone resorption observed in MM. PMID- 11918538 TI - Enhanced antitumoral effectiveness of idiotype vaccination induced by the administration of Flt3 ligand combined with interleukin 2 against a murine myeloma. AB - Idiotype (Id) vaccination provides an innovative treatment modality against B cell malignancies. In multiple myeloma patients, however, the antitumoral potential of this immunotherapeutic concept is limited. In an attempt to improve the therapeutic effectiveness of Id vaccination, we added Flt3 ligand (Flt3-L) and interleukin 2 (IL-2) to the protocol. Balb/c mice were inoculated i.p. (d -2) with different doses (1-5 x 10(5)) of HOPC myeloma cells, secreting the IgHOPC Id protein. Two days later, animals were treated with Flt3-L (10 microg per mouse/d, given i.p) for 10 consecutive days (d 0-9). On d 5 and d 11, myeloma-specific immunoglobulin (Ig(HOPC)) was administered s.c., together with incomplete Freund adjuvans (IFA) followed by the administration of IL-2 (2 x 10.000/d given i.p) for 10 d (d 5-14). Whereas Ig(HOPC), Flt3-L or IL-2, given alone, did not elicit long-term survival, the combination of IL-2 or Flt3-L with Id vaccination achieved a complete tumour rejection in 27% and 41% of mice respectively. However, the most powerful antimyeloma effects were induced by Flt3-L + Id vaccination + IL-2: 81% of the treated animals experienced long-term survival (> 180 d). Both natural killer (NK) cells and CD8+ T cells may be involved in the antitumoral immune response. These data suggest that the combination of Flt3-L and IL-2 can be used to enhance the therapeutic effectiveness of clinical cancer vaccination protocols. PMID- 11918539 TI - Myeloma of the central nervous system: association with high-risk chromosomal abnormalities, plasmablastic morphology and extramedullary manifestations. AB - Involvement of the central nervous system (CNS) by multiple myeloma, as defined by the detection of malignant plasma cells in the cerebrospinal fluid in the presence of suggestive symptoms, is considered extremely rare. We report on the characteristics of 18 such patients diagnosed and treated at the University of Arkansas over the last 10 years for an overall incidence of approximately 1%. Their evaluation revealed association of CNS involvement with unfavourable cytogenetic abnormalities (especially translocations and deletion of the chromosome 13), high tumour mass, plasmablastic morphology, additional extramedullary myeloma manifestations and circulating plasma cells. The presence of these features should alert clinicians to the possibility of CNS involvement. The outcome of these patients was extremely poor despite the use of aggressive local and systemic treatment including autologous stem cell transplants. Given this universally poor prognosis, the application of allogeneic transplants should be studied in this clinical setting. PMID- 11918540 TI - Influence of karyotype on outcome of allogeneic bone marrow transplantation for adults with precursor B-lineage acute lymphoblastic leukaemia in first or second remission. AB - The prognostic relevance of karyotype has been established in adult acute lymphoblastic leukaemia (ALL) patients treated with chemotherapy but not definitively evaluated in an allogeneic bone marrow transplantation (BMT) setting. To determine the factors affecting the outcome of allogeneic BMT for adults with precursor B-lineage ALL in first or second complete remission (CR), a total of 41 consecutive patients with a successful karyotype were enrolled in this study. There were 21 men and 20 women with a median age of 27 (15-43) years. The distribution of French-American-British (FAB) subtypes was as follows: L1 (n = 26), L2 (n = 15). Unfavourable karyotypes (n = 12) were defined as Ph+ or t(4;11). Disease status at the time of transplant was first CR (n = 35) or second CR (n = 6). With a median follow-up of 36 months, the 3-year probabilities of relapse and disease-free survival (DFS) were 36.3 +/- 8.4% and 57.3 +/- 8.4% respectively. Potential variables predicting worse relapse and DFS were FAB subtype (L2), extramedullary involvement, pre-BMT status (second CR), unfavourable karyotype and type of graft-versus-host disease (GVHD). Further multivariate analysis showed that karyotype and pre-BMT status were independently associated with relapse and DFS. In addition, chronic GVHD was found to be significantly associated with a lower relapse rate. PMID- 11918542 TI - Epstein-Barr virus-associated T-cell acute lymphoblastic leukaemia. AB - Epstein-Barr virus (EBV) is frequently detected in haematological malignancies, including Burkitt's lymphomas/leukaemias, Hodgkin's diseases and non-Hodgkin's lymphomas. However, immature T-cell malignancies associated with EBV have not been reported previously. We report a patient with T-cell acute lymphoblastic leukaemia (T-ALL), whose leukaemic cells had EBV, confirmed by Southern blotting and in situ hybridization. The EBV existed in episomal form and was detected in most leukaemic cells, but not in bystander normal B-cells. The leukaemic cells, massively infiltrated into the liver and spleen, were resistant to chemotherapy. EBV might be associated with tumorigenesis of T-ALL, and characteristic clinical features of the patient. PMID- 11918541 TI - A pilot study of the recombinant soluble human tumour necrosis factor receptor (p75)-Fc fusion protein in patients with myelodysplastic syndrome. AB - Laboratory observations suggest that, in some myelodysplastic syndromes (MDS), immune mechanisms may contribute to the impaired blood cell production. Tumor necrosis factor alpha (TNF-alpha), a potent inhibitor of haematopoiesis, has been hypothesized to mediate suppressive effects in MDS: TNF-alpha levels are elevated and correlated with marrow apoptosis and cytopenia. Inhibition of TNF-alpha production using the soluble TNF receptor (Enbrel) has been successful in rheumatoid arthritis, and we have now applied the same principle to MDS. We determined spontaneous TNF-alpha production by marrow cells in MDS; TNF-alpha production was elevated (> mean + 2 x SD of controls) in > 1/3 of patients, but did not correlate with clinical parameters. Sixteen patients participated in a 3 month pilot study of Enbrel. The drug was well tolerated and 15 patients were evaluable. Of these, one became temporarily (14 weeks) transfusion independent. In another patient, absolute neutrophil count (ANC) rose from 0.5 x 10(9)/l to 0.84 x 10(9)/l. Serious infections were seen in two out of six neutropenic patients. Progression to refractory anaemia with excess blasts in transformation (RAEBt) or leukaemia was observed in three patients. When the effects of Enbrel on haematopoietic colony formation were studied, no significant increase was seen in MDS and there was no correlation with TNF-alpha levels. Although anti-TNF therapy with Enbrel was well tolerated at the dosages used in MDS, its efficacy as a single agent appears low. PMID- 11918543 TI - Combined arsenic trioxide and all-trans retinoic acid treatment for acute promyelocytic leukaemia recurring from previous relapses successfully treated using arsenic trioxide. AB - The optimal treatment of acute promyelocytic leukaemia (APL) recurring from relapses successfully treated using arsenic trioxide (As2O3) is undefined. Three APL patients relapsing from As2O3-induced remission were studied. Re-treatment with As2O3 failed in one patient in third relapse, and resulted in morphological but not molecular remission in another patient. Combination therapy with As2O3 and all-trans retinoic acid (ATRA), however, resulted in morphological and molecular remission in all three cases, with a follow-up time ranging from 6 to 16 months. Our results suggest a synergistic therapeutic effect between As2O3 and ATRA in APL in advanced relapse. PMID- 11918544 TI - Fluoro-deoxyglucose positron emission tomography imaging for the detection of occult disease in multiple myeloma. AB - Positron emission tomography with 2-deoxy-2-[18]fluoro-D-glucose (FDG-PET) imaging has been extensively used to detect occult metastatic malignant lesions in patients with carcinoma. We describe its use in three patients with multiple myeloma, each representing a particular clinical situation in which this imaging modality offered advantages over plain radiography, computerized tomography or magnetic resonance imaging. FDG-PET provides a whole body image showing sites of occult disease. This is of particular value in patients with non-secretory myeloma, solitary plasmacytoma or for those that relapse with focal disease following autologous or allogeneic stem cell transplantation. PMID- 11918545 TI - Factor VIII inhibitor in a patient with mild haemophilia A and an Asn618-->Ser mutation responsive to immune tolerance induction and cyclophosphamide. AB - We describe a patient with mild haemophilia A (original value of factor VIII activity 0.30 U/ml) who developed an inhibitor (36.1 Bethesda U/ml) which cross reacted with his endogenous factor VIII. This caused a decline in basal factor VIII level (< 0.01 U/ml) and severe haemorrhagic events. Treatment to induce immune tolerance was started with factor VIII/von Willebrand factor (VWF) concentrates, but inhibitor levels increased progressively and the patient suffered serious bleeding. Cyclophosphamide was administered and, after 8 months treatment, factor VIII levels increased to 0.20 U/ml and the inhibitor could no longer be detected. Screening of his factor VIII gene revealed a missense mutation in exon 13 that predicts substitution of Asn618-->Ser in the A2 domain of factor VIII. Immunoprecipitation analysis showed that the antibodies present in the patient's plasma reacted with metabolically labelled A2 domain and, to a lesser extent, with factor VIII light chain. Inhibitory antibodies were completely neutralized by recombinant A2 domain, whereas no neutralization was observed after the addition of factor VIII light chain (A3-C1-C2) and C2 domain. More detailed analysis showed that the majority of inhibitory antibodies were directed against residues Arg484-Ile508, a previously identified binding site for factor VIII inhibitors. Our findings suggest that immune tolerance therapy and cyclophosphamide were successful in eradicating inhibitory antibodies against a common epitope on factor VIII. PMID- 11918546 TI - Semiquantitative measurement of IgG subclasses and IgM of platelet-specific antibodies in a glycoprotein-specific platelet-antigen capture assay. AB - The detection of platelet-specific antibodies is of high clinical interest in diseases with immune thrombocytopenia. The glycoprotein-specific platelet-antigen capture (PAC)-assay developed in this study is especially suited to the differentiation of platelet-specific immunoglobulin (Ig) G subclasses and the determination of platelet-specific IgM in serum or on platelets. The problems with unspecific signals or low sensitivity usually seen with the detector antibodies available are effectively overcome, as unbound detector antibodies are removed at an early stage in the assay. We investigated 14 maternal alloantisera from cases of neonatal alloimmune thrombocytopenia (NAITP) and six sera from patients with autoimmune thrombocytopenic purpura (AITP). In NAITP sera, we found IgG1 alone in 57%, IgG1 + IgG3 in 21% and IgG1 + IgG2 in 14% of cases. One serum contained IgG1 + IgG2 + IgG3. In AITP, three out of the six sera contained IgG1 alone. One serum contained IgG1 + IgG2. One patient, with highly refractory AITP, had platelet-specific IgG1 + IgG2 + IgG3 in his serum. A patient with AITP in remission and normal platelet counts only showed platelet-bound IgG2. The detection of platelet-specific 'whole IgG' is possible too. However, at this time the commonly used monoclonal antibody-specific immobilization of platelet antigens (MAIPA) method should not be replaced for this purpose, as it is well standardized and used with similar results in many laboratories. The PAC assay sensitively detects the subclasses of platelet-specific IgG and human leucocyte antigen (HLA)-antibodies independently. It is easy to perform and takes less time than other platelet glycoprotein-specific methods. PMID- 11918548 TI - Age, sex and vitamin status affect plasma level of homocysteine, but hyperhomocysteinaemia is possibly not an important risk factor for venous thrombophilia in Taiwanese Chinese. AB - The biological effects of age, sex and vitamin status on plasma total homocysteine (tHcy), and association of hyperhomocysteinaemia with venous thromboembolism in Taiwanese Chinese individuals, were investigated. Eighty patients (16-85 years) with venous thrombophilia and 123 healthy subjects (15-85 years) without history of vascular thrombosis were studied for plasma levels of tHcy, folate and vitamin B12. A multivariate analysis in healthy subjects revealed that plasma tHcy levels tended to increase with age (P < 0.001) and with decreasing plasma levels of folate (P=0.001) or vitamin B12 (P < 0.029); men tended to have higher plasma tHcy levels than women (P=0.006). Thrombotic risk assessment in a case-control study demonstrated that neither plasma level of tHcy [odds ratio (OR), 1.07; 95% confidence interval (CI), 0.96-1.18; P=0.210] nor hyperhomocysteinaemia (OR, 1.65; 95% CI, 0.50-5.49; P=0.415) was significantly associated with venous thrombophilia. The relationship between hyperhomocysteinaemia and recurrence of episode remained insignificant (P=0.560). We conclude that age, sex and vitamin status affect plasma tHcy but hyperhomocysteinaemia is possibly not an important risk factor for venous thrombophilia in Taiwanese Chinese. PMID- 11918547 TI - Regulation of plasminogen activator inhibitor-1 secretion by urokinase and tissue plasminogen activator in rat epithelioid-type smooth muscle cells. AB - Tissue plasminogen activator (tPA) and urokinase (uPA) are targets of plasminogen activator inhibitor-1 (PAI-1) inhibition. We have previously shown that both proteases can also induce PAI-1 secretion in rat smooth muscle cells (SMCs). We now report that both proteases appear to use very similar cellular mechanisms for signal transduction. They induced PAI-1 secretion using a pathway(s) involving protein kinase C (PKC). They also activated the Raf/Mek/mitogen-activated protein kinase (MAPK) pathway, which lies downstream of PKC activation. Activation of protein kinase A (PKA), however, lowered PAI-1 secretion induced by uPA and tPA, as a result of an inhibition of the PKC pathway and inhibition of Raf, Mek and MAPK phosphorylations. Src and syk family non-receptor tyrosine kinases (TK) were also involved in PAI-1 induction. The mechanisms of interaction of these tyrosine kinases with other pathways appeared to be quite different: src appeared to act within the PKC and PKA pathways, while syk operated independently of these pathways. Furthermore, whereas src inhibition resulted in inhibition of Raf/Mek/Erk phosphorylations, syk inhibition could only inhibit Mek and Erk phosphorylations but not the phosphorylation of Raf. These multiple pathways utilized by uPA and tPA to modulate PAI-1 secretion might be involved in determining the proteolytic or antiproteolytic potential of the SMCs under different pathophysiological conditions. PMID- 11918549 TI - Immunocytochemistry for the heavy chain of the non-muscle myosin IIA as a diagnostic tool for MYH9-related disorders. AB - May-Hegglin anomaly (MHA), Sebastian syndrome (SBS) and Fechtner syndrome (FTNS) are autosomal-dominant macrothrombocytopenias with Dohle-like leucocyte inclusions. These diseases are due to mutations of the MHY9 gene, encoding the heavy chain of non-muscle myosin IIA (NMMHC-A). We investigated the NMMHC-A localization in blood cells from eight MHA, SBS or FTNS patients with known MYH9 mutations. All the patients showed an altered localization of NMMHC-A in granulocytes and platelets, suggesting that Dohle-like bodies are due to the aggregation of NMMHC-A in the cytoplasm. Therefore, immunocytochemistry for NMMHC A is a simple and sensitive method to detect pathological phenotypes of granulocytes and platelets in the diagnosis of MYH9-related disorders. PMID- 11918550 TI - Characterization of two novel splice site mutations in human factor VII gene causing severe plasma factor VII deficiency and bleeding diathesis. AB - The molecular basis of severe type I factor (F)VII deficiency was investigated in two Algerian patients. One patient, a 13-year-old-girl who has suffered from severe bleeding since birth, was homozygous for a 7-bp deletion (nt 7774-7780) and a 251-bp insertion (nt 7773-7781) of mitochondrial origin, in IVS 4 acceptor splice site. The other patient, an infant who died from massive intracranial haemorrhage, was homozygous for a transversion in the IVS 7 donor splice site (T9726+2-->G) and a missense mutation in exon 8 (G10588-->A; Arg224-->Gln). In both cases, the deleterious mutations are probably the splice site junction abnormalities impairing mRNA processing. These three lesions have not yet been reported. PMID- 11918551 TI - Inherited factor VII deficiency and surgery: clinical data are the best criteria to predict the risk of bleeding. AB - Inherited factor VII (FVII) deficiency is a rare autosomal disorder characterized by a weak relationship between FVII activity (FVII:C) and operative bleeding risk. We report a retrospective study of 17 patients with a FVII:C below 0.1 IU/ml, in whom surgery was performed without any replacement therapy. Clinical and biological data were analysed to establish predictive criteria for bleeding tendency. We found that systematic preoperative replacement therapy may not be necessary for 'minor' surgical procedures, for patients suffering from inherited FVII deficiency, unless the clinical history includes severe haemorrhagic symptoms such as haemarthrosis, severe haematomas (even of soft tissue) or abundant epistaxis. PMID- 11918552 TI - Reversion of autoimmune lymphoproliferative syndrome with an antimalarial drug: preliminary results of a clinical cohort study and molecular observations. AB - Autoimmune lymphoproliferative syndrome (ALPS) is a paediatric disease characterized by lymphoproliferation and autoimmunity. Most patients are known to carry heterozygous mutations of the TNFRSF6 gene leading to diminished Fas mediated apoptosis and failure of activated lymphocytes to undergo apoptosis. A subgroup of patients without the TNFRSF6 gene mutation has similar defective apoptosis and clinical features. No effective treatment has been reported so far. Glucocorticoids, intravenous immunoglobulin and/or immunosuppressive drugs have usually led to only transient clinical improvement. Seven ALPS patients (two type Ia and five type III) were treated with the antimalarial drug Fansidar. No toxicity was observed. An objective response was seen in six of them and, in two, the treatment was stopped without reappearance of the symptoms. Moreover, a marked decrease in interleukin-10 levels was observed in two patients during the treatment. We found that the drug induced apoptosis in activated lymphocytes through activation of the mitochondrial apoptotic pathway. PMID- 11918553 TI - Abnormal telomere shortening in leucocytes of children with Shwachman-Diamond syndrome. AB - Haemopoietic dysfunction, ranging from single-lineage cytopenia to severe aplasia and/or myelodysplasia (MDS), is prominent in Shwachman-Diamond syndrome (SDS). To assess haemopoietic stem cell proliferation in SDS, we compared leucocyte telomere length in 12 patients with SDS to that of 41 controls, using an in-gel hybridization technique. SDS patients had an age-adjusted mean telomere length 1.4 kilobase pairs (kbp) shorter than controls (P < 0.0001). Patients with'non severe' SDS (one- or two-lineage cytopenias; no MDS) had shortened telomeres ( 1.4 kbp; P = 0.0004), as did those with 'severe' SDS. We conclude that stem cell hyperproliferation is a feature of SDS from its outset. PMID- 11918554 TI - Enhanced haemolysis with beta-thalassaemia trait due to the unstable beta chain variant, Hb Gunma, accompanied by hereditary elliptocytosis due to protein 4.1 deficiency in a Japanese family. AB - We identified a Japanese family with a beta-thalassaemia trait and hereditary elliptocytosis (HE). We studied five members of this family. One was normal, one had only the beta-thalassaemia trait, one had heterozygous HE, and two had compound heterozygous beta-thalassaemia trait and HE. The last two had already undergone splenectomy. The molecular profile of beta-thalassaemia was consistent with that of Hb Gunma: codon 127/128CAGGCT(Gln-Ala)--> CCT(Pro). Analysis of erythrocyte membrane proteins revealed a partial deficiency of protein 4.1 in all those with HE, whereas the spectrin content was within the normal range. Each heterozygous family member with either the beta-thalassaemia trait or HE was asymptomatic, whereas the two with both beta-thalassaemia and HE had marked red blood cell deformities and haemolysis. The abnormalities of the red blood cells in patients with the beta-thalassaemia trait might be enhanced by association with HE owing to a protein 4.1 deficiency. PMID- 11918555 TI - Gastroduodenal lesions in polycythaemia vera: frequency and role of Helicobacter pylori. AB - The prevalence of gastroduodenal lesions is higher in polycythaemia vera (PV) than in the general population. However, the role of Helicobacter pylori (H. pylori) in the pathogenesis of such lesions is unknown. The aim of our study was to evaluate the prevalence of gastroduodenal lesions in PV patients and dyspeptic controls, and to assess the role of PV and H. pylori infection in inducing them. Thirty-five PV patients fulfilling selection criteria and 73 age- and sex-matched controls underwent upper gastrointestinal endoscopy. Six gastric mucosal biopsies were taken in all patients and controls, and analysed for presence of H. pylori; serum anti-CagA was assayed by Western blot. Data were analysed with descriptive statistics and multivariate regression analysis. Compared with controls, PV patients showed a significantly higher frequency of erosions (46% versus 12%), ulcers (29% versus 7%), H. pylori positivity (83% versus 57%), and anti-CagA positivity (66% versus 37%). Fourteen out of 20 (70%) asymptomatic PV patients had gastroduodenal lesions. At multivariate analysis, H. pylori, presence of PV alone, and both PV and anti-CagA were significantly and strongly associated with a higher frequency of gastroduodenal lesions (P < 0.05, P < 0.01 and P < 0.05 respectively). Both PV and H. pylori infection were independent risk factors for gastroduodenal lesions; the underlying pathogenetic mechanism responsible for gastroduodenal lesions in PV possibly involves blood mucosal flow and trophism. The higher susceptibility of H. pylori infection and the high frequency of asymptomatic gastroduodenal lesions in PV patients suggest a surveillance of these patients. PMID- 11918556 TI - Assignment of functional roles to parasite proteins in malaria-infected red blood cells by competitive flow-based adhesion assay. AB - Adhesion of parasitized red blood cells (PRBCs) to endothelial cells and subsequent accumulation in the microvasculature are pivotal events in the pathogenesis of falciparum malaria. During intraerythrocytic development, numerous proteins exported from the parasite associate with the RBC membrane skeleton but the precise function of many of these proteins remain unknown. Their cellular location, however, suggests that some may play a role in adhesion. The adhesive properties of PRBCs are best studied under flow conditions in vitro; however, experimental variation in levels of cytoadherence in currently available assays make subtle alterations in adhesion difficult to quantify. Here, we describe a flow-based assay that can quantify small differences in adhesion and document the extent to which a number of parasite proteins influence adhesion using parasite lines that no longer express specific proteins. Loss of parasite proteins ring-infected erythrocyte surface antigen (RESA), knob-associated histidine-rich protein (KAHRP) or Plasmodium falciparum erythrocyte membrane protein 3 (PfEMP3) had a significant effect on the ability of PRBCs to adhere, whereas loss of mature parasite-infected erythrocyte surface antigen (MESA) had no effect. Our studies indicate that a number of membrane skeleton-associated parasite proteins, although not exposed on the RBC surface, can collectively affect the adhesive properties of PRBCs and further our understanding of pathophysiologically relevant structure/function relationships in malaria infected RBCs. PMID- 11918557 TI - Familial pseudohyperkalaemia Cardiff: a mild version of cryohydrocytosis. AB - We have investigated a Welsh pedigree showing the 'familial pseudohyperkalaemia' phenotype of dominantly inherited, red-cell-based, temperature-dependent pseudohyperkalaemia associated with normal haematology. The 'passive leak' to K across the membrane of these abnormal red cells showed a 'U-shaped' temperature dependence, with a minimum at about 23 degrees C, qualitatively similar to that seen in the frankly haemolytic 'cryohydrocytosis' variant of the hereditary stomatocytosis group. Like three previous pedigrees with cryohydrocytosis, these patients show an excess of ether lipids in the membrane. However, these patients differ from other 'familial pseudohyperkalaemia' pedigrees, in which the leak showed different temperature profiles. PMID- 11918558 TI - An estimate of the current risk of transmitting blood-borne infections through blood transfusion in Italy. AB - We conducted a retrospective cohort study to estimate the incidence of major blood-borne agents among Italian blood donors and calculated the risk of infection among blood recipients using the 'incidence/window period model'. The study was conducted among 46 180 blood donors enrolled in six blood centres between 1994 and 1999. During follow-up, seven new infections were confirmed: three donors seroconverted for anti-human immunodeficiency virus (HIV); two for anti-hepatitis C virus (HCV); and two showed hepatitis B surface antigen (HBsAg) reactivity; no cases of syphilis were observed. The incidence rates per 100 000 person/years were: 4.06 (95% CI: 0.82-11.85) for HIV; 2.41 (95% CI: 0.29-8.70) for HCV; and 2.70 (95% CI: 0.32-9.77) for HBsAg; the incidence for total hepatitis B virus (HBV) infection was 9.77 per 100 000 person/years (95% CI: 1.16 35.36). The estimated risk of an infectious blood unit not being detected was: 2.45 (95% CI: 0.13-12.33) per 1 million units for HIV; 4.35 (95% CI: 0.30-22.39) for HCV; and 15.78 (95% CI: 1.16-84.23) for HBV. Overall, an estimated 22.58 per 1 million units are infected. In Italy, the risk of transfusion-transmitted infections is low and is similar to that in other western countries. The introduction of new more sensitive screening tests could reduce the residual risk of transfusion-transmitted infection by 40-80%. PMID- 11918559 TI - Heterozygosity for two novel null alleles of the KEL gene causes the Kell-null phenotype in a Japanese woman. AB - The Kell-null (Ko) phenotype is rare and it does not express the Kell antigens on erythrocyte membranes. Recently, several distinct missense and nonsense base substitutions in the coding region and the donor splice site of intron 3 were identified in the KEL gene in individuals with the Ko phenotype. We analysed both genomic DNA and cDNA sequences of the KEL gene in a Japanese woman with the Ko phenotype. She was found to be heterozygous for two novel null KEL alleles. One allele contained an A to G substitution in intron 5 that changes the 3'-splice site of intron 5 from AAG to AGG, resulting in a reading frameshift by a single guanine insertion in KEL mRNA, and the other allele contained a single G to A substitution in exon 12 (codon 459) creating a termination codon. Neither mutation was found in 114 randomly selected Japanese individuals. The results suggested that the Ko blood group phenotype might be owing to several distinct non-functional alleles without any prevalent allele. PMID- 11918560 TI - Selective in vitro expansion and efficient retroviral transduction of human CD34+ CD38- haematopoietic stem cells. AB - Ex vivo expansion of primitive human haematopoietic stem cells (HSC) is clinically relevant for stem cell transplantation and gene therapy. Here, we demonstrate the selective expansion of CD34+CD38- cells from purified CD34+ cells upon stimulation with Flt3-ligand, stem cell factor and thrombopoietin. Over a 100-fold (range 80 to 128-fold) expansion of CD34+CD38- cells was observed with bone marrow and cord blood (CB). The expanded CD34+CD38- cells remained negative for lineage-specific markers and could be induced to differentiate into granulocytes, monocytes, megakaryocytes, erythrocytes, and T and B-lymphocytes in vitro. Lineage differentiation assays with single CD34+CD38- cells showed no loss of multilineage potential of expanded cells after ex vivo culture. We also demonstrated that the increase in frequency of CD34+CD38- cells was not as a result of the downregulation of CD38 expression during the culture. Quantitative analysis showed that the number of 6 week cobblestone area forming cells (CAFCwk6), a measure of proliferating HSC, in cytokine-stimulated CD34+ cells were increased by 20-fold. Expanded CD34+CD38- cells could be transduced efficiently with retroviruses encoding the low affinity nerve growth factor receptor (LNGFR) marker gene (17% to 44%, mean 27%), resulting in long-lasting expression of retroviral-encoded genes in progeny HSC and differentiated progenitors. We conclude that the combination Flt3-ligand (FL), stem cell factor and thrombopoietin (TPO) induced strong ex vivo proliferation of CD34+CD38- cells and that the absolute number of expanded cells with stem cell activity increased substantially in this population. PMID- 11918562 TI - Syngeneic stem cell transplant for spent-phase polycythaemia vera: eradication of myelofibrosis and restoration of normal haematopoiesis. AB - We report a patient with spent-phase polycythaemia vera (S-PV) and massive splenomegaly who failed to engraft after a syngeneic granulocyte colony stimulating factor-primed peripheral blood stem cell transplant (SCT), but later engrafted after splenectomy. Bone marrow (BM) showed resolution of myelofibrosis (MF) and absent endogenous erythroid colonies. This case demonstrated that (1) normal haematopoiesis can be restored after syngeneic SCT despite extensive MF, and (2) fibrosis can regress following a total body irradiation-containing regimen and syngeneic SCT. As a graft-versus-BM stroma effect is non-existent in syngeneic transplants, there may be a role for autologous SCT to obliterate MF in S-PV. PMID- 11918561 TI - CD34+/CD90+ cells infused best predict late haematopoietic reconstitution following autologous peripheral blood stem cell transplantation. AB - This study aimed to identify which graft product subset of cells might be the most predictive of late haematopoietic recovery (three to 12 months) following autologous peripheral blood stem cell transplantation (PBSCT). The relationships between the numbers of reinfused CD34+ cells and their immature subsets such as CD34+/CD90+, CD34+/AC133+, CD34+/CD38- and CD34+/HLA-DR- cells, and haemoglobin, white blood cell (WBC) and platelet counts at 3, 6, 9 and 12 months after PBSCT, were studied in 25 patients with haematological and solid malignancies. The total CD34+ cell number, as well as CD34+/CD90+ and CD34+/AC133+ cell numbers, correlated with platelet counts at 3, 6, 9 and 12 months after PBSCT, but the CD34+/CD90+ cells infused best predicted platelet recovery during the first 12 months after PBSCT (P < 0.0238 at any time-point). The CD34+/AC133+ cell dose also correlated with WBC counts at 3 months post PBSCT. In addition, all patients receiving more than 80 x 10(4) CD34+/CD90+ cells/kg showed platelet counts greater than 100 x 10(9)/l at all points after PBSCT, suggesting that this value of the CD34+/CD90+ cells infused was a threshold dose for durable haematopoietic engraftment after PBSCT. PMID- 11918563 TI - A survey of one-stage and chromogenic potencies in therapeutic factor VIII concentrates. PMID- 11918564 TI - Complete, long-term remission of refractory idiopathic cold haemagglutinin disease after Mabthera. PMID- 11918565 TI - A case of severe refractory thrombotic thrombocytic purpura responding to treatment with 13-cis retinoic acid. PMID- 11918567 TI - Phenotyping primitive plasma cells. PMID- 11918566 TI - Chronic relapsing thrombotic thrombocytopenic purpura due to a deficiency of von Willebrand factor-cleaving protease activity. PMID- 11918568 TI - Levels of von Willebrand factor-cleaving protease are normal in methylene blue treated fresh-frozen plasma. PMID- 11918569 TI - Acceptance rates and publication times. PMID- 11918570 TI - Dentistry between pathology and cosmetics. AB - In ancient and medieval times, the prevalence of caries and periodontal disease varied. There were no treatments for dental hard tissue, but dental cosmetics played an important role. In the late 19th century, caries levels reached a maximum in Europe and North America after refined sugar became a cheap staple food. Toothlessness became frequent even in young adults. Caries prevention, effective on a public health scale, began with the introduction of water fluoridation in the 1940s. By 1985, dental academia had acknowledged that substantial declines could also be obtained in entire populations through topical fluorides, mainly in toothpastes. While decreasing caries prevalence is irrefutable in affluent countries, the specific reasons of the decline are still a matter of debate. In countries where caries has declined substantially, activities of dentists are shifting towards cosmetic dentistry. However, caries continues to be a problem for the lower socioeconomic strata, even in affluent countries, and is a serious problem in developing countries. Thus, water fluoridation is still important, and salt fluoridation should be considered where water fluoridation is not feasible. Both measures are extremely cheap to implement. Controlled fluoridation has a great potential for developing countries and low social strata of affluent countries. Its reduced effectiveness in high socioeconomic strata of affluent countries, due to the widespread usage of fluoride in toothpastes and other oral care products, should not detract from the public health value of fluoridation. PMID- 11918571 TI - Erosion, caries and rampant caries in preschool children in Jeddah, Saudi Arabia. AB - OBJECTIVES: The objective of this study was to determine the prevalence of dental erosion in preschool children in Jeddah, Saudi Arabia, and to relate this to caries and rampant caries in the same children. METHODS: A sample of 987 children (2-5 years) was drawn from 17 kindergartens. Clinical examinations were carried out under standardised conditions by a trained and calibrated examiner (M.Al-M.). Measurement of erosion was confined to primary maxillary incisors and used a scoring system and criteria based on those used in the UK National Survey of Child Dental Health. Caries was diagnosed using BASCD criteria. Rampant caries was defined as caries affecting the smooth surfaces of two or more maxillary incisors. RESULTS: Of the 987 children, 309 (31%) had evidence of erosion. For 186 children this was confined to enamel but for 123 it involved dentine and/or pulp. Caries were diagnosed in 720 (73%) of the children and rampant caries in 336 (34%). The mean dmft for the 987 children was 4.80 (+/-4.87). Of the 384 children who had caries but not rampant caries, 141 (37%) had erosion, a significantly higher proportion than the 72 (27%) out of 267 who were clinically caries free (SND=2.61, P<0.01). Of the 336 with rampant caries, 96 (29%) also had evidence of erosion. CONCLUSIONS: The level of erosion was similar to that seen in children of an equivalent age in the UK. Caries was a risk factor for erosion in this group of children. PMID- 11918572 TI - Follow-up study of dental students' esthetic perceptions of mild dental fluorosis. AB - OBJECTIVES: Several studies have assessed people's esthetic perceptions of dental fluorosis, suggesting that concerns may be greater than believed previously. Few studies have assessed dental students' perceptions and none has done so over time. The purpose of this paper is to report on changes in dental students' esthetic perceptions of dental fluorosis and other conditions. METHODS: Fourth year dental students (n=45) completed questionnaires about computer-generated photographs of fluorosis and other conditions, using the same protocol as when they were entering dental students. Results were compared for each of the eight images at the individual level using paired statistical tests. RESULTS: Although many patterns were generally consistent, there were a substantial number of differences in results over time, concerning both the mild fluorosis and non fluorosis images. In general, students tended to score both fluorosis and non fluorosis images more favorably as fourth-year students. When rating images from very pleased (1) to very embarrassed (10), fourth-year students rated the images significantly more favorably than they had done as first-year students for normal/control, incisal third only dental fluorosis, and a more generalized, mild fluorosis. Similarly, with a visual analog scale from satisfactory (0) to unsatisfactory (69), fourth-year students reported significantly lower (more favorable) scores than they had done as first-year students for the normal/control, diastema/no fluorosis, more involved mild fluorosis, and incisal third only mild fluorosis. There were no obvious changes in the relative levels of favorable evaluation of fluorosis vs. the other conditions. CONCLUSIONS: Fourth-year dental students generally had more favorable esthetic perceptions of mild dental fluorosis and other conditions than they had reported as entering students. PMID- 11918573 TI - Tooth loss in the very old: 13-15-year incidence among elderly Iowans. AB - OBJECTIVES: Very few studies have reported tooth loss incidence over a period of 10 years or more, and fewer have reported tooth loss occurrence in subjects aged 80 and older, so that the long-term pattern of tooth loss in the very old is largely unknown. This study assessed 13-15-year tooth loss incidence among a cohort of Iowans, aged 65 and older at baseline. METHODS: Oral examinations were conducted on 520 subjects beginning in 1983, and periodically until 1988, with another round of examinations conducted on surviving members of the initial cohort during 1996-98. RESULTS: Of the 73 remaining subjects, 45 subjects lost a total of 153 teeth during the period (mean=2.1 teeth lost), with a maximum of 17 teeth lost. Molars were the most commonly lost teeth, while canines and maxillary incisors were the least commonly lost. Bivariate analyses found that tooth loss was associated with untreated decay at baseline and level of periodontal attachment loss at earlier examinations. Logistic regression identified only greater severity of attachment loss as a significant risk factor for tooth loss (Adjusted odds ratio=2.4, P=0.006). The impact of tooth loss on subjects' lives was assessed using OHIP and other questions. The occurrence of tooth loss over the study period had little impact, but the number of remaining teeth significantly impacted subjects' ability to eat or chew food, swallow, or their willingness to smile. CONCLUSIONS: These results suggest that tooth loss continues in the very old, that periodontal attachment loss is associated with tooth loss in this age group, and that loss of teeth over one's lifetime does affect certain quality-of-life measures. PMID- 11918574 TI - Risk factors for dental erosion in 5-6 year old and 12-14 year old boys in Saudi Arabia. AB - MATERIAL AND METHODS: Dental examinations were carried out on 354 boys aged 5-6 years, and 862 boys aged 12-14 years, attending 40 schools in Riyadh. The prevalence of dental erosion was assessed using diagnostic criteria similar to those employed in the 1993 UK National Survey of Child Dental Health. RESULTS: Pronounced dental erosion (into dentine or dentine and pulp) was observed in 34% of 5-6 year olds and 26% of 12-14 year olds. Information on food and drink consumed and dietary habits was obtained by means of a questionnaire. Parents reported that 65% of 5-6 year old boys took a drink to bed. Water was the commonest drink consumed (37%) followed by carbonated soft drinks (21%). One third of parents reported that their son had something to eat in bed or during the night and 60% of this was sweet food or confectionery. Seventy per cent of 12 14 year old boys reported consuming drinks at night; these were mainly water (30%), carbonated soft drinks (27%) and tea or coffee, with sugar (18%). Forty six per cent of the 12-14 year olds reported that they ate in bed at least once a week and 54% of this was sweet food or confectionery. When the dental examination and questionnaire results were correlated, a statistically significant relationship was found between the number of primary maxillary incisors with pronounced erosion of their palatal surfaces and the consumption of carbonated soft drinks at night (P=0.015). A significant relationship was also found between the number of permanent maxillary incisors with pronounced erosion on their palatal surfaces and the frequency of drinks at night (P=0.020), as well as the duration of drinks retained in the mouth (P=0.038). CONCLUSION: It is concluded that dental erosion is more common in the primary and permanent dentitions of Saudi Arabian boys compared with results for similar age groups from the United Kingdom. PMID- 11918575 TI - A longitudinal study of occlusal caries among schoolchildren in Dar es Salaam, Tanzania. AB - OBJECTIVES: To describe the clinical caries status and the radiographic progression of occlusal caries lesions in permanent first and second molars among primary schoolchildren in Dar es Salaam over a 3-year period. METHODS: Clinical and radiographic diagnosis of caries in first and second permanent molars in 223 children aged 8-16 years were carried out annually from 1994 to 1997. The drop out rates from the baseline in 1994 to the follow-up examinations in 1995, 1996 and 1997 were 16.6%, 22.0% and 35.4%, respectively. Twenty-one percent of the dropouts were picked up during the study. RESULTS: Less that 5% of all occlusal surfaces that were sound at the beginning of the study developed new clinical caries lesions over the 3-year study period. The highest rate of new lesions was found in second molars. The mandibular second molars were most frequently affected by new caries lesions followed by the mandibular first molars. Progression of lesions was generally slow. After 1, 2 and 3 years, 30.0%, 47.9% and 52.8% of lesions in occlusal surfaces of first molars had progressed, compared to 47.9%, 71.3% and 100.0% of lesions in second molars. CONCLUSIONS: Dental caries prevalence was low. New occlusal lesions were more likely to appear in mandibular second molars. Carious lesions were progressing slowly, especially in the first molars. Fissure abrasion may play a role in minimizing the risk of developing new occlusal lesions as well as progression of existing lesions. PMID- 11918576 TI - Oro-facial pain in the community: prevalence and associated impact. AB - STUDY OBJECTIVE: To determine the prevalence of oro-facial pain (OFP) in the population and within-population subgroups and to describe the associated disability. DESIGN: Cross-sectional population study. SETTING: General medical practice in South East Cheshire, United Kingdom. PARTICIPANTS: A random sample of 4000 adults aged 18-65 years of whom 2504 responded (adjusted participation rate 74%). MAIN RESULTS: The overall prevalence of OFP was 26% (95% Confidence Interval (CI) 24%, 28%). The prevalence of symptoms was higher in women (30%) than in men (21%) and in both sexes the highest (30%) prevalence was found in the 18-25 year age group and the lowest (22%) in the 56-65 age group. Of all the participants, 12% had pain in or around the eyes, 10% reported pain in and around the temples, 6% pain in front of the ears and 6% pain in the jaw joints. Only 46% of the participants with OFP had sought professional advice from a dentist or general medical practitioner and 17% had to take time off work or were unable to carry out normal activities because of pain. CONCLUSIONS: OFP is a common symptom experienced by a quarter of the adult population, of whom only 46% seek treatment. The prevalence is higher in women and younger age groups. PMID- 11918577 TI - Plaque minerals in the prediction of caries activity. AB - BACKGROUND/AIMS: Attempts to identify subjects who will develop caries lesions in future years have been only moderately successful, suggesting that one or more important risk factors are being overlooked. The aim of this study was to validate the use of plaque calcium, phosphate and fluoride concentrations as significant risk factors in caries. METHODS: DMFS and DMFT rates were measured three times over 2 years in a group of rural Chinese schoolchildren initially aged 12 years and not living in a high-fluoride area. At the baseline and year 1 examinations, dental plaque was accumulated for 3 days and, after collection, was analysed for Ca, P and F. Plaque scores, dental health behaviour and parents' occupation data were also recorded. RESULTS: The mean DMFS increment over 2 years was 1.14 with a range of -2 to +9. Regression analysis taking all factors into account identified only plaque Ca concentration, baseline DMFS score and toothbrushing frequency as significant factors in identifying high-risk individuals. Plaque Ca also showed predictive ability on its own. For example, when high caries was defined as 3+ new DMFS and a Ca cut-off value of 200 nmol/mg dry wt was selected, sensitivity was 0.84 and specificity 0.38. However, with this cut-off value the percentage predicted to have high caries risk was unrealistically high at 66%. As in several other studies, baseline caries score was a useful predictor of future caries. CONCLUSION: Low plaque Ca concentration showed a modest ability to predict future caries, and since it is implicated directly in the chain of events leading to caries, its use in caries prediction should be considered further. Plaque P and F showed no predictive ability in this study. PMID- 11918579 TI - Allergic contact dermatitis: the cellular effectors. AB - Contact hypersensitivity reactions are mediated by lymphocytic effector cells. Until recently it was believed that the most important of these were CD4+ T lymphocytes. However, there is growing evidence that in many instances the predominant effector cell may be a CD8+ T lymphocyte, with in some instances CD4+ cells performing a counter-regulatory function. Here we review the roles of CD4+ T helper (Th) cells and CD8+ T cytotoxic (Tc) cells, and their main functional subpopulations (respectively, Th1 and Th2 cells and Tc1 and Tc2 cells) in the elicitation of contact hypersensitivity reactions and consider the implications of effector cell selectivity for the biology of allergic contact dermatitis. PMID- 11918578 TI - A blind caries and fluorosis prevalence study of school-children in naturally fluoridated and nonfluoridated townships of Morayshire, Scotland. AB - OBJECTIVES: To undertake a blind caries and fluorosis prevalence study of Grade 1 (aged 5/6 yr) and Grade 4-7 (aged 8-12 yr) children from naturally water fluoridated (1 ppm, since 1985) Burghead, Findhorn & Kinloss (F), and nearby nonfluoridated Buckie & Portessie (N-F), in rural Morayshire, Scotland. METHODS: A blind clinical (+ 10% repeats) caries study of the above townships' 5/6-yr-old lifetime (15 F; 43 N-F), and 8-12-yr-old lifetime (55 F; 136 N-F)/school-lifetime (31 F; 37 N-F) residents was undertaken following bussing of these children to a common examination site in close-by Elgin Town Hall. Initially, each child was asked about their own perception of the aesthetics of their maxillary front teeth. Fluorosis was assessed clinically using the TF Index, as well as photographically - for later blind scoring (+ 10% repeats for lifetime 8-12-yr olds) of slides by four dental and two lay 'jurors', alongside a now-established UK 'bench-mark' mildly mottled (TFI = 2), fluorosis comparator slide, judged in previous studies to be aesthetically lay-acceptable. In addition, by parental questionnaire, information was sought concerning their child's fluoride supplement and dentifrice usage histories. RESULTS: For 5/6-yr-olds, mean primary caries scores were 96.0% less in fluoridated than nonfluoridated subjects (P < 0.01). In 8-12-yr-olds, DMFT values favoured water-fluoridated subjects; their caries-free trend was significant (P < 0.001 overall). Clinically, 33% of all lifetime F subjects and 18% of all N-F pupils had fluorosed maxillary anterior teeth (P = 0.045), but no statistically significant difference was found between the 7% F and 3% N-F subjects with TFI scores > 2 (P = 0.25). Photographically, 'jury' mottling assessment (+ 10% repeats) of projected slides resulted in at least 1 : 6 positive scores in 43.6% of F and 30.9% of N-F pupils, albeit they unanimously scored only nine F and five N-F children as having fluorosed teeth (P < 0.01). In no case did all members score TFI > 2. Dental and lay scorers rated TFI = (1/2) in only a further 9.1% and 5.5% of F subjects, respectively, compared to 0.7% and 1.5% respectively of N-F pupils. Again, TFI > 2 was scored unanimously in no child. No differences were found regarding the children's own degree of anterior tooth aesthetic nonacceptability between F (11%) and N-F (12%) prevalence (P = 0.75). Finally, only one F child had taken F supplements and, while 26 N-F had used F drops, no significant relationship was found between their usage and TFI values in the latter group (P = 0.49). Additionally, no relationship was noted between clinical TFI scores and the age at which parents stated fluoridated dentifrice toothbrushing commenced, between 0 and 24 + months of age. CONCLUSIONS: Considerable caries benefit has accrued to those Morayshire rural children who have received naturally fluoridated water (at 1 ppm) throughout their lives, as compared to their socioeconomically similar, nonfluoridated rural counterparts. Furthermore, in spite of all but two subjects claiming to have brushed regularly with fluoridated dentifrice (and no evidence of the availability of nonfluoridated toothpaste being purchasable in the five townships), only borderline mild fluorosis disadvantages have been noted clinically, and none by the subjects' own aesthetic perceptions. Finally, no evidence was found to suggest any delay in permanent tooth eruption patterns of the F subjects. It would seem appropriate therefore, that adjustment of Scots' drinking waters' natural fluoride levels to 1 ppm should be pursued to extend similar dental advantages to the vast majority of that population (both young and old) which, it is well documented, has the worst dental health of mainland UK. PMID- 11918580 TI - Natural vegetable fats in the prevention of irritant contact dermatitis. AB - Chronic irritant contact dermatitis (ICD) is one of the most pressing problems in occupational medicine and is common in the food processing industry. To date, protective creams that fulfil the special requirements in the foodstuffs industry have not been available. Therefore, we studied the efficacy of pre-exposure application of natural vegetable fats in the prevention of experimentally induced ICD. A panel of 20 healthy volunteers was tested with a repetitive irritation test using sodium lauryl sulfate (SLS) as a standard irritant in a randomized study. Application sites were assessed clinically and by the use of bioengineering techniques (evaporimetry, chromametry, and corneometry). Rape seed and palm fats showed significant protective potential. Gas-chromatographic analysis revealed differences in the fatty acid composition of the vegetable. Higher content of linoleic acid and lower content of oleic acid was associated with beneficial effects. Our results are a new approach in the prevention of ICD and towards the development of new protective preparations for workplaces in the foodstuffs industry. PMID- 11918581 TI - A case of contact urticaria syndrome due to di(2-ethylhexyl) phthalate (DOP) in work clothes. AB - We previously reported a case of contact urticaria syndrome (CUS) due to di(2 ethylhexyl) phthalate (DOP) in a polyvinyl chloride (PVC) grip on cotton gloves. The patient reported in this previous paper was careful not to have any contact with PVC products in his daily life or in his working environment. He discontinued the use of protective gloves with a PVC grip that was the cause of CUS. When working, he used cotton gloves without a PVC grip. We prescribed antihistamines which slightly improved his condition. However, when he wore work clothes while on duty, CUS relapsed. This condition was severe and made him feel anxious. When we advised him to wear a cotton shirt under his work clothes, the contact urticaria did not develop. We suspected that some component of the work clothes was the cause of his symptoms. A prick test with the extract solution of his work clothes showed a wheal and flare at the 15 min reading. The common component of the grip and the work clothes was found by analysis to be DOP. PMID- 11918582 TI - Patch test results in 542 patients with suspected contact dermatitis in Turkey. AB - In this retrospective study, patch test results of 542 patients (303 female and 239 male), referred to our clinic between June 1996 and July 1999, were evaluated. All patients were patch tested with 32 allergens of an extended European standard series, 140 patients were tested with supplemental series, and 246 patients with their own substances. 280 (51.7%) patients had 1 or more positive results but allergic contact dermatitis, according to clinical relevance, was diagnosed in only 190 (67.9%) of these. Nickel sulfate was the most frequent sensitizer (19.1%), followed by potassium dichromate (11.8%), palladium chloride (9.4%), cobalt chloride (8.5%), and thiuram mix (7.7%). Frequencies of contact allergy to fragrance mix (5.5%), balsam of Peru (Myroxylon Pereirae resin) (2%), quaternium-15 (0.6%), and Kathon CG (methylchloroisothiazolinone+methylisothiazolinone) (0.2%) were relatively low, while no positive reactions were obtained to paraben mix in any case. Current clinical relevance to the present dermatitis was most frequently seen with carba mix (96.2%), thiuram mix (95.2%) and potassium dichromate (78.1%), whereas this rate was low for nickel (37.5%). In 85.8% of patients with allergic contact dermatitis, the responsible allergen was detected by testing with the extended standard series alone (18.4% identified by testing with the additional 10 allergens), in 11.6% by testing with the patients' own substances, and in 2.6% by testing with the supplemental series. Occupational contact dermatitis was diagnosed in 77 of the patients with positive reactions (27.5%), most of these being construction workers and house painters who showed relevant sensitizations to potassium dichromate, cobalt chloride, thiuram mix and carba mix. There was no statistically significant difference in the total contact sensitization rate between atopics and non-atopics, but contact sensitivity to nickel sulfate and palladium chloride was significantly higher among atopics. MOAHL index in % was: 44.1/14.2/24.9/ 54.4/2.4. In conclusion, we believe that similar large series of patients would be helpful to delineate the exact profile of contact dermatitis in Turkey. PMID- 11918583 TI - Biophysical changes after mechanical injury of the stratum corneum in normal skin. AB - Scrubbing off the stratum corneum with a rough towel after soaking in warm water is a bathing custom unique to Korea. However, Korean dermatologists have advised against this practice due to the potential harm that it may cause, though there is little data to support this advice. This study aimed to evaluate the changes in biophysical characteristics after such mechanical injury of stratum corneum, as reflected by water-holding capacity, stratum corneum barrier function, and stratum corneum turnover time (SCTT). 10 Korean female volunteers (aged 19-34 years) were enrolled in this study to investigate the acute effects and 32 volunteers (aged 19-49 years) the chronic effects. After soaking in warm water (36 degrees C) for 5 min, the flexor surfaces of the forearms were scrubbed with a rough towel. The volunteers' forearms were divided into 4 compartments, and each compartment was scrubbed 0, 5, 10, and 15x, respectively. After a single injury of the stratum corneum, there was an initial decrease in water-holding capacity until 6 h. Thereafter, it increased until day (D) 3, when it began to descend to the normal range. The transepidermal water loss (TEWL) values also peaked at D2 and declined thereafter. However, there were no changes in either water-holding capacity or skin barrier function, and only the SCTT was significantly shortened, after chronic injury of the stratum corneum. PMID- 11918584 TI - Arthus reaction to lepirudin, a new recombinant hirudin, and delayed-type hypersensitivity to several heparins and heparinoids, with tolerance to its intravenous administration. AB - The pathogenesis of allergic reactions to heparin is poorly understood. Clinically, this phenomenon is relevant because of its increasing incidence and the resulting therapeutic challenges due to various cross-reactions between unfractionated and low-molecular weight heparins as well as between heparins and heparinoids. A 44-year-old female patient had developed a delayed-type hypersensitivity to certoparin-sodium. Diagnostic allergy testing revealed various cross-reactions between different heparins as well as an intolerance to heparinoids. After subcutaneous challenge with the recombinant hirudin lepirudin (Refludan) the patient developed a local Arthus reaction at the injection site. In general, recombinant hirudins do not cross-react with high- or low-molecular weight heparins and heparinoids because of a different molecular structure and are therefore an alternative in case of adverse reactions to heparins and heparinoids. Whereas a local Arthus reaction has already been described twice for low-molecular weight heparins, this is to the best of our knowledge the first observation of a superficial leukocytoclastic vasculitis due to s.c. applied lepirudin. Intravenous administration of heparins and heparinoids in case of hypersensitivity to these drugs following topical application risks a generalized eczematous reaction in patients with delayed-type allergy to both groups of substances. In our patient with delayed-type hypersensitivity to heparins and heparinoids and superficial vasculitis due to lepirudin, the intravenous challenge with heparin and a heparinoid was justified as an ultima ratio measure and proved to be the useful therapeutical alternative. PMID- 11918585 TI - Patch testing for permanent-press allergic contact dermatitis. AB - During the decade of the 90s, 24 cases of permanent-press textile-resin contact dermatitis were identified. The most common allergen associated with these cases was dimethylol propylene urea (DMPU). Although previous reports had suggested ethylene urea melamine formaldehyde resin (EUMF) or dimethylol dihydroxy ethylene urea (DMDHEU) as the most appropriate test materials for this type of contact dermatitis, we could not confirm EUMF as appropriate but, with some limitations, can support DMDHEU due to its widespread and continued use. DMPU has been withdrawn from the marketplace and replaced with what may be less reactive materials. In this study, we also observed discordance between EUMF patch tests in the range of 30%, which is consistent with previous reports about the biovariability of patch test results. PMID- 11918586 TI - Different physical forms of maleopimaric acid give different allergic responses. AB - Endo-maleopimaric acid (MPA) is a contact allergen formed when colophonium is "modified" with maleic anhydride or fumaric acid. Previous patch testing showed a higher allergic response to petrolatum (pet.) preparations of MPA in amorphous form compared to MPA in crystalline form. In the present study, the impact of the physical form of MPA on the allergic response was investigated. Since the amorphous form is difficult to standardize, crystalline MPA mechanically incorporated or dissolved in pet. was used. A lower eliciting capacity was obtained from crystalline MPA, compared to that obtained from dissolved MPA, in guinea pigs intradermally induced with MPA. Using 3H-MPA a 3X difference in the dissolution into synthetic sweat from MPA dissolved in pet., compared to MPA mechanically incorporated, was demonstrated. A difference in bio-availability between dissolved and crystalline MPA could therefore be assumed. Crystalline MPA had a low sensitizing capacity compared to that seen for amorphous MPA in previous studies. The amorphous form of MPA is likely to have a larger surface area than crystalline MPA, with less ordered molecules, resulting in a higher dissolution rate and a greater bio-availability. Modified colophonium exists as amorphous solids and as viscous liquids. Thus, exposure will probably be to non crystalline MPA and cases of contact allergy could be overlooked when screening with crystalline MPA. PMID- 11918587 TI - Adverse effects of gasoline on the skin of exposed workers. AB - Gasoline is widely used as a solvent in industry. To study its adverse effects on the skin and to understand their mechanisms, a matched epidemiological study (1:1, 52 exposed workers and 52 control subjects) was developed. Information about general conditions, history of dermatosis, changes in skin after exposure to gasoline, etc., was obtained. Ceramide, fatty acid and cholesterol collected from the backs of the hands were analyzed by high-performance thin-layer chromatography (HPTLC), because stratum corneum lipids play a predominant role in maintaining the physiological function of skin. The results showed that prevalences of hyperkeratosis, dryness, onychosis and dermatitis were clearly higher in exposed workers than in the control group, prevalence ratios being 3.33 (p<0.05), 3.00 (p<0.001), 11.25 (p<0.001), 5.00 (p<0.001), respectively. Fissures and onychorrhexis were the common symptoms in exposed workers. The stratum corneum lipid levels of ceramide, fatty acid and cholesterol were significantly lower in the exposed group than in the control group (p<0.05). Findings indicated that prolonged or repeated contact with gasoline could cause fissuring of the skin and nail disorders, and that the mechanism was perhaps depletion of stratum corneum lipids. PMID- 11918588 TI - 'Lucky Luke' contact dermatitis from diapers with negative patch tests. PMID- 11918589 TI - Suitability of the European standard series of patch test allergens in Pakistani patients. PMID- 11918590 TI - Contact urticaria from geraniol. PMID- 11918591 TI - Airborne irritant contact dermatitis from phosphates in a fertilizer factory. PMID- 11918592 TI - Severe contact urticaria and anaphylaxis from benzophenone-3(2-hydroxy 4-methoxy benzophenone). PMID- 11918593 TI - Allergic contact dermatitis from dibutyl phthalate and benzalkonium chloride in Timodine cream. PMID- 11918594 TI - Allergic contact dermatitis due to methyl ethyl ketone peroxide, cobalt naphthenate and acrylates in the manufacture of fibreglass-reinforced plastics. PMID- 11918595 TI - Allergic contact dermatitis from 2-bromo-2-nitropropane-1,3-diol in Metrogel. PMID- 11918596 TI - Allergic contact dermatitis from unsaturated polyester resin consisting of maleic anhydride, phthalic anhydride, ethylene glycol and dicyclopentadiene. PMID- 11918597 TI - Allergenicity of natural rubber latex gloves. AB - The concentrations of proteins, allergens and rubber chemicals are essential parameters of the allergenicity of NRL gloves. To date, a standardized method has been given only for analysis of the protein concentration (DIN EN 455-3), and not for the concentrations of allergens and rubber chemicals. In the present study, we investigated 11 brands of surgical gloves currently available on the German market. Additionally 1 glove, not subjected to final leaching procedures, was added for comparison purposes. Protein concentrations were analysed by different methods in different laboratories. Allergen concentrations were assayed by prick tests in NRL-allergic volunteers and by RAST inhibition methods. Rubber chemicals were analysed by HPTLC and GC. The protein concentrations analysed by the Lowry method in the 2 laboratories gave concordant results, but the correlations between protein and allergen concentrations were low. The protein concentration analysed by HPLC correlated with the allergen concentration, and gave better information on the allergenicity of the gloves. The development of standardized methods for allergen analysis in the assessment routine is necessary, due to significant discrepancy between protein and allergen levels of some gloves. Thiurams were not found in any of the gloves, though carbamates were present in all gloves tested. Our data indicate that washing procedures have little or no effect on the concentration of rubber chemicals. PMID- 11918598 TI - Occupational protein contact dermatitis from spices in a butcher: a new presentation of the mugwort-spice syndrome. AB - Protein contact dermatitis to meat is well known in butchers; spices are another source of potential contact allergy and usually are not recognized. We present a first case of contact-dermatitis to spice mix in a 39-year-old-butcher. The patient underwent skin prick testing (SPT) with standard allergens (ALK) and different meat and spice extracts (Stallergenes), scratch-patch testing with spice mix containing glutamate, paprika and other spices. Specific serum-IgE was measured with CAP-FEIA. SPT only showed an immediate-type sensitization to mugwort (+ +), as well as different spices (paprika +, curry +, cumin +) and camomile (+ + +). Scratch-patch tests were negative for different meat, but strongly positive for spice mix (+ + +) after 30 min (wheal and flare) and (+ +) after 48 h (infiltration and vesiculation). Two healthy controls were tested negative for spice mix used from that patient (scratch-patch). Specific IgE was slightly elevated for paprika 0.47 kU/L (CAP class 1), anise 0.43 kU/L, curry 0.36 kU/L and mugwort 3.83 kU/L. Sx1 atopy-multiscreen was 3.8 kU/L due to a sensitization to mugwort alone. The tests performed demonstrate an IgE-mediated contact allergy to spices but also a delayed type allergy to spice mix as a manifestation of the mugwort-spice syndrome in this individual. When testing for occupational dermatitis in butchers, protein contact allergy to spices must also be taken into consideration. PMID- 11918599 TI - Effects of repeated short-term skin contact with proteolytic enzymes. AB - The objective was to investigate effects on the hand of protease used in dishwashing liquids (DWL). A group of 30 Koreans (normal skin) and 24 Japanese (12 atopic and 12 normal) were employed for two different studies, respectively. The 30 Koreans were divided into three groups. Each group immersed their hands for 15 min a day for 9 days into DWL containing 0.005% protease, 0.02% protease, or non-enzyme DWL. The 24 Japanese immersed their hands for 15 min a day for 4 days into 0.005% protease-containing or a non-enzyme DWL. The hand skin was evaluated by measuring overall and dry skin grades, estimated before and after exposures to the test products. The Korean study shows that 0.005% and 0.02% protease-containing DWL are less irritating to the skin than non-enzyme control, as measured by better overall skin grades and less dry skin. The Japanese study shows that 0.005% protease-containing DWL are less irritating to the skin than non-enzyme control in both atopic and normal subjects. The 0.005% protease use did not cause any adverse dermatological effects to atopic subjects compared with non-enzyme control. Both studies indicate that 0.005% and 0.02% protease resulted in better skin mildness profile, without causing adverse dermatological complications to the subjects. PMID- 11918600 TI - Skin protection in bakers' apprentices. AB - Skin protection measures - barrier creams, protective gloves - and skin care are widely recommended for the prevention of occupational hand dermatitis (HD) in skin risk professions, but there is hardly anything known about uptake levels of the measures. The objective of this controlled intervention study was to quantify the uptake and maintenance of skin protection and skin care measures in first year bakers' apprentices. A total of 94 first-year bakers' apprentices were included in the study in September 2000. The apprentices were assigned to the skin protection and control group class-wise to reduce contamination. The skin protection group comprised 39 apprentices who were trained in skin protection measures at the beginning and after 4 weeks of training. 55 apprentices were assigned to the control group representing no skin protection intervention. Standardized interviews took place at the beginning of the training and at 4 monthly follow-ups (FU). The uptake of skin protection measures differed significantly between the groups (barrier cream p < 0.0001, protective gloves p = 0.046, skin care p = 0.025). Barrier cream use in the skin protection group was incorporated in the daily routine very well from the start and reached 100% at the end of the examination period (4th FU). At this time, only 3.2% of the controls used barrier creams. The level of acceptance of protective gloves (4th FU: skin protection group 43.3%; controls 32.3%) was considerably lower than that of barrier creams. The initial level of regular skin care was high in both groups (skin protection group 67.6%, controls 61.7%). After the intervention the acceptance of skin care rose to 88.9% in the skin protection group compared to 68.1% in the controls (4th FU). The present study has shown that skin protection and skin care measures can be introduced successfully in the daily routine of a skin risk occupation and high uptake and maintenance rates can be achieved. PMID- 11918601 TI - Population differences in acute skin irritation responses. Race, sex, age, sensitive skin and repeat subject comparisons. AB - The variability in human skin irritation responses has been well documented and can confound our ability to accurately assess differences in skin reactivity between human subpopulations. In the current analysis, results were compiled from nine acute irritation patch test studies, conducted at three test facilities over a 5-year period. Four irritant test chemicals, 20% sodium dodecyl sulphate, 100% decanol, 100% octanoic acid and 10% acetic acid, were tested in sufficient numbers of test subjects to enable the stratification of results for different human subpopulations. An increased reactivity was noted for Asian versus Caucasian subjects for each of three test chemicals, in contrast to the previously described individual study results from which these data were drawn. Male subjects were directionally or significantly more reactive to each of the test chemicals than female subjects. The oldest age cluster of subjects (56-74 years of age) was directionally or significantly less reactive than younger age clusters. There was virtually identical reactivity between self-assessed 'sensitive' and normal skin groups. Lastly, there was little correlation between the results from individual subjects tested in two or more studies with the same chemicals. These results add to our general understanding of population differences in skin reactivity and the potential implications for ingredient and product skin safety testing and risk assessment. PMID- 11918602 TI - Type IV hypersensitivity to vitamin K. AB - The day after intramuscular injection of vitamin K1 (phytomenadione) into her thigh, a 27-year-old-woman with normal liver function developed a relapsing and remitting eczematous reaction localized to the injection site, and later a further eczematous reaction under an adhesive dressing (Duoderm). On patch testing, she was positive to vitamin K1 and cross-reacted to vitamin K4; she was also positive to colophonium and to ester gum rosin, the dressing adhesive. Recurrent angioedema persisted for several months and, 2 years later, symptoms were still occurring over the injection sites. Structure-activity relationships among vitamin K allergens are discussed. PMID- 11918603 TI - Relationship between 2,4-dinitrochlorobenzene elicitation responses and individual irritant threshold. AB - Contact dermatitis shows significant interindividual variation. We studied the relationship between an individual's irritant threshold and the elicitation of experimentally induced allergic sensitization. The results suggest that there is an association between reactivity to an irritant and the likelihood of positive elicitation reactions to lower hapten concentrations. This novel finding would be compatible with the proposal that contact allergy is best explained using a danger model with cutaneous irritancy determining sensitization. However, the enhanced allergic reactivity of individuals with a lower irritant threshold was not apparent on visual inspection, suggesting that the determinants of skin reactivity to haptens are multifactorial. PMID- 11918604 TI - Occupational irritant and allergic contact dermatitis among healthcare workers. AB - Contact dermatitis is the most frequent occupational dermatosis and non-specific irritants in addition to specific Type IV sensitization are involved. We reviewed our database for data from 1994 to 1998 and selected 360 consecutive patients working in healthcare environments and experiencing contact dermatitis at their hands, wrists and forearms. We found that allergic contact dermatitis and irritant contact dermatitis were considered to be work-related in 16.5% (72/436) and 44.4% (194/436) of diagnoses, respectively. Occupational irritant contact dermatitis is due to exposure to a wide range of irritants in the workplace, such as soaps, solvents, cleansers and protective gloves, which conspire to remove the surface lipid layer and/or produce cellular damage. In this study the major relevant aetiological agents that induced occupational allergic contact dermatitis were: nickel sulphate (41 patch positivities), components of disinfectants [glutaraldehyde (5) and benzalkonium chloride (7)] and rubber chemicals [thiuram mix (15), carba mix (9) and tetramethylthiuram monosulphide (6)]. The best treatment for allergic contact dermatitis is to avoid those allergens causing the rash. Whenever this is not possible, contact with them needs to be reduced using properly selected protective gloves. Finally, subjects with atopic dermatitis should avoid 'wet work' and contact with irritants, because atopic dermatitis is significantly associated with irritant contact dermatitis. PMID- 11918605 TI - Contact dermatitis from the essential oil of tangerine in fragrance. PMID- 11918606 TI - Allergic contact dermatitis from undecylenic acid in a commercial antifungal nail solution. PMID- 11918607 TI - Baboon syndrome from 5-aminosalicylic acid. PMID- 11918608 TI - Contact nummular (discoid) eczema from depilating cream. PMID- 11918609 TI - Anaphylaxis after hamster bites: a rare case? PMID- 11918610 TI - Lichenoid reaction from a permanent red tattoo: has nickel a possible aetiologic role? PMID- 11918611 TI - Occupational allergic contact dermatitis and Type I sensitization caused by Gerbera in a greenhouse worker. PMID- 11918613 TI - Frequency of textile dye and resin sensitization in patients with contact dermatitis in Israel. PMID- 11918612 TI - Clinical relevance of + patch test reactions to lanolin alcohol. PMID- 11918614 TI - Contact allergy to diglycolamine in a water-based metalworking fluid. PMID- 11918615 TI - Fixed drug eruption caused by metronidazole. PMID- 11918616 TI - Is the application of cosmetics containing protein-derived products safe? PMID- 11918617 TI - Systemic contact dermatitis from diallyl disulfide. PMID- 11918618 TI - Contact dermatitis from Hedera helix in a husband and wife. PMID- 11918619 TI - Contact urticaria from hops (Humulus lupulus) in a patient with previous urticaria-angioedema from peanut, chestnut and banana. PMID- 11918620 TI - Gastric emptying in diabetes: clinical significance and treatment. AB - The outcome of recent studies has led to redefinition of concepts relating to the prevalence, pathogenesis and clinical significance of disordered gastric emptying in patients with diabetes mellitus. The use of scintigraphic techniques has established that gastric emptying is abnormally slow in approx. 30-50% of outpatients with long-standing Type 1 or Type 2 diabetes, although the magnitude of this delay is modest in many cases. Upper gastrointestinal symptoms occur frequently and affect quality of life adversely in patients with diabetes, although the relationship between symptoms and the rate of gastric emptying is weak. Acute changes in blood glucose concentration affect both gastric motor function and upper gastrointestinal symptoms. Gastric emptying is slower during hyperglycaemia when compared with euglycaemia and accelerated during hypoglycaemia. The blood glucose concentration may influence the response to prokinetic drugs. Conversely, the rate of gastric emptying is a major determinant of post-prandial glycaemic excursions in healthy subjects, as well as in Type 1 and Type 2 patients. A number of therapies currently in development are designed to improve post-prandial glycaemic control by modulating the rate of delivery of nutrients to the small intestine. PMID- 11918621 TI - Assessing diabetic control--reliability of methods available in resource poor settings. AB - AIMS AND METHODS: To examine the reliability of random venous or capillary blood glucose testing, random urine glucose testing, and a current symptom history in predicting a high HbA1c in Type 2 diabetic patients taking oral hypoglycaemic agents in a poorly controlled rural African population. RESULTS: For a cut-off point for HbA1c of > or = 8%, for random venous plasma glucose of > or = 14 mmol/L (present in 47.2% of subjects), specificity was 97.1% (95% CI 85.1-99.9), sensitivity 56.8% (48.8-64.5) and positive predictive value (PPV) 98.9% (94.2 99.9). HbA1c > or = 8% is predicted by a random capillary blood glucose of 17 mmol/L (present in 28.4% of subjects) with specificity 100% (90.0-100.0), PPV 100% (93.7-100.0) and sensitivity of 34.3% (27.2-42.1). HbA1c > or = 8% is predicted by the presence of heavy glycosuria (> or = 55 mmol/L) (present in 35.6%) with specificity 94.1% (80.3-99.3), sensitivity of 41.9% (34.1-49.9) and PPV 97.1% (89.9-99.6). Polyuria/nocturia (present in 31.3%) was the only symptom found to be associated with poor control, with a specificity for predicting HbA1c of > or = 8% of 81.5% (61.9-93.7), PPV 89.1% (76.4-96.4) and sensitivity 30.6% (22.9-39.1). CONCLUSIONS: Where resources are short, random glucose testing can be used to detect a significant proportion of those with the worst control with a high degree of specificity enabling primary care staff to modify treatment safely. Where facilities are limited capillary blood or urine testing with reagent strips, may be substituted for venous plasma testing in the laboratory. A symptom history was insufficient to replace biochemical testing, but where this is unavailable, urinary symptoms may be helpful. PMID- 11918622 TI - Design of the Collaborative AtoRvastatin Diabetes Study (CARDS) in patients with type 2 diabetes. AB - BACKGROUND: There are few data on the role of lipid lowering in the primary prevention of coronary heart disease (CHD) in diabetic patients. This paper describes the design of a collaborative clinical trial between Diabetes UK, the NHS Research and Development Directorate and Pfizer UK, that addresses this question. METHODS: The Collaborative AtoRvastatin Diabetes Study (CARDS) is a multicentre, randomized, placebo-controlled, double-blind clinical trial of primary prevention of cardiovascular disease in patients with Type 2 diabetes. The primary objective is to investigate whether treatment with the hydroxymethylglutaryl coenzyme A reductase inhibitor, atorvastatin, reduces the incidence of major cardiovascular events. At entry patients have at least one other risk factor for CHD in addition to diabetes, namely current smoking, hypertension, retinopathy, or micro- or macroalbuminuria. At randomization patients have been selected for a serum low-density lipoprotein (LDL) cholesterol concentration < or = 4.14 mmol/l (160 mg/dl) and triglycerides < or = 6.78 mmol/l (600 mg/dl). Randomization was completed in June 2001. Patients will be followed until 304 primary endpoints have accrued (expected date early 2005). The trial includes 2838 men and women aged 40-75 years. This report describes the design and administration of the study and reviews the evidence to date of the effectiveness of lipid-lowering therapy in Type 2 diabetes. CONCLUSIONS: The case for lipid-lowering therapy for the primary prevention of CHD in diabetes has not been demonstrated. CARDS will provide essential information on the extent of any benefits and adverse effects of lipid-lowering therapy in diabetic patients without prior CHD. PMID- 11918623 TI - Paraoxonase gene cluster is a genetic marker for early microvascular complications in type 1 diabetes. AB - BACKGROUND: Paraoxonase is a serum enzyme, which prevents oxidation of low density lipoprotein (LDL) by hydrolyzing lipid peroxides. Two polymorphisms in PON1 gene have been associated with cardiovascular and microvascular diseases in both diabetic and non-diabetic patients. AIMS: The current project was designed to investigate the association between the polymorphisms of two PON genes and diabetes microvascular diseases (retinopathy and microalbuminuria) and any potential linkage between Met54Leu of PON1 and Cys311Ser of PON2 gene. METHODS: Diabetic retinopathy and albumin excretion rate were assessed in 372 adolescents with Type 1 diabetes who were genotyped for the two polymorphisms. RESULTS: We confirmed the increased susceptibility for diabetic retinopathy for the Leu/Leu genotype (odds ratio (OR) 3.34 (confidence interval (CI) 1.95, 5.75), P < 0.0001). The Ser/Ser genotype was significantly more common in those patients with microalbuminuria (albumin excretion rate > or = 20 microg/min) compared with those with albumin excretion rate < 20 microg/min (OR 4.72 (CI 2.65, 8.41), P < 0.0001). The Ser311 of PON2 was in strong linkage disequilibrium with Leu54 of PON1 gene (Delta = 23 x 10(4), P < 0.001). The delta value was higher for those without complications (28 x 104, P < 0.001) compared with those with complications (15.5 x 10(4), P < 0.001). CONCLUSIONS: This study supports the hypothesis that diabetic microangiopathy is genetically heterogeneous. PON1 Leu/Leu increases the risk for retinopathy and PON2 Ser/Ser increases the risk for microalbuminuria. PMID- 11918624 TI - St Vincent's Declaration 10 years on: outcomes of diabetic pregnancies. AB - AIMS: To monitor pregnancies in women with pregestational Type 1 diabetes for pregnancy loss, congenital malformations and fetal growth parameters, in a geographically defined area in the north west of England. METHODS: Population cohort study of 547 pregnancies in women with Type 1 diabetes from maternity clinics in 10 centres over a 5-year period (1995-1999 inclusive). Main outcome measures were numbers and rates of miscarriages, stillbirths, neonatal and post neonatal deaths; prevalence of congenital malformations; birth weight in relation to gestational age. RESULTS: Among 547 pregnancies, there were six (1.1%) pairs of liveborn twins, 439 (80.3%) liveborn singletons; 72 (13.2%) spontaneous abortions, 14 (2.6%) stillbirths and 16 (2.9%) terminations. Four of the terminations were performed because of congenital malformations. Both the stillbirth rate (30.1/1000 total births (95% confidence interval (CI) 16.6 50.0)), and prevalence of congenital malformations (84.3/1000 live births (95% CI 60.3-113.8)) were significantly higher than the local population (P < 0.001). When corrected for gestational age, mean birth weight in the sample was 1.3 sd greater than that of infants of non-diabetic mothers (P = 0.12). Infants with congenital malformations weighed less than those without. CONCLUSION: In an unselected population, the infants of women with pregestational Type 1 diabetes mellitus have 6.4 times the reported risk of a congenital malformation and 5.1 times the reported risk of perinatal mortality than infants in the general population. Further improvements in the management of diabetes and pregnancy in these women are needed if the St Vincent's Declaration target is to be met. PMID- 11918625 TI - Clinical characteristics, GAD antibody (GADA) and change of C-peptide in Korean young age of onset diabetic patients. AB - AIMS: To investigate the association of clinical and immunological markers with diabetes classification in newly diagnosed young diabetic patients at disease onset. METHODS: Eighty-two diabetic patients recruited within 1 year of onset (mean age 23.0 +/- 7.1, M:F = 46:36) were divided into three groups, namely: a low fasting C-peptide (FC) level at baseline group (the low FC group (n = 14, FC < 0.18 nmol/l)), an intermediate FC group (n = 29, 0.18 nmol/l < or = FC < 0.37 nmol/l), and a high FC group (n = 39, 0.37 nmol/l < or = FC). Patients were reclassified at follow-up (mean follow-up period 3.7 +/- 1.4 years) in the same manner as described above into low FC group (n = 31), intermediate FC group (n = 20), and high FC group (n = 31). The clinical characteristics and prevalence of GADA were compared. RESULTS: Patients in the high FC group at baseline had a higher body mass index (BMI), a higher frequency of a family history of diabetes, a higher meal-stimulated C-peptide increment, a lower frequency of ketonuria, a lower frequency of history of diabetic ketoacidosis, and a lower frequency of insulin therapy at diagnosis than those in the low and intermediate FC groups at baseline. Insulin secretory capacity, which was represented by fasting C-peptide, was affected by BMI at diagnosis and the presence of GADA. All the patients of the low FC group on follow-up were finally classified as having Type 1 diabetes; moreover, the factors that determined the type of diabetes were lower BMI at onset, GADA positivity, insulin therapy, lower fasting C-peptide level and lower meal-stimulated C-peptide increment at initial admission. CONCLUSIONS: Our study suggests that follow-up involving C-peptide and GADA measurements in combination with clinical characteristics is useful for discriminating between the types of diabetes in these groups. PMID- 11918626 TI - eNOS4 polymorphism of the endothelial nitric oxide synthase predicts risk for severe diabetic retinopathy. AB - BACKGROUND: Genetic factors may be involved in the development, and particularly in the severity, of diabetic retinopathy (DR), in addition to chronic hyperglycaemia. Increased nitric oxide generation has been suggested to play a significant role in the pathogenesis of DR. AIMS AND METHODS: To examine whether the eNOS4 is involved in the risk of severe DR, 200 unrelated Caucasian Type 1 diabetic patients of long duration were randomly selected (M/F 103/97, age 44.4 +/- 12.4 years, diabetes duration 27.7 +/- 10.0 years, body mass index 24.3 +/- 3.4 kg/m2, HbA1c 8.6 +/- 1.3%). The eNOS4 polymorphism was analysed by polymerase chain reaction, and DR by retinal angiography and classified as presence (n = 101) or absence (n = 99) of severe (proliferative or pre-proliferative) DR. RESULTS: The genotype distribution of eNOS4b/b (wild-type), eNOS4b/a (heterozygous) and eNOS4a/a (homozygous) was 72%, 24.5% and 3.5%, respectively. Frequency of eNOS4a/a was significantly lower in patients with severe DR (n = 0) when compared with controls (n = 7, odds ratio (OR) = 0 (95% confidence interval (CI) = 0.5-0.74), P = 0.02). eNOS4b/b was more frequent in patients with severe DR (n = 80) when compared with controls (n = 64, OR = 2.1 (95% CI = 1.1-4.12), P = 0.032). Frequency of eNOS4b/a was not different between the study (n = 21) and control groups (n = 28, ns). The allelic frequencies between the study and control groups were different (4b: n = 181 vs. n = 156, respectively, OR = 2.3 (95% CI = 1.27-4.25), P = 0.005; 4a: n = 21 vs. n = 42, respectively, OR = 0.4 (95% CI = 0.24-0.79), P = 0.005). CONCLUSIONS: We demonstrate in Caucasians with Type 1 diabetes that (i) eNOS4a/a is associated with absent or non-severe DR, and (ii) eNOS4b/b is associated with severe DR. PMID- 11918627 TI - Resource consumption and costs in Dutch patients with type 2 diabetes mellitus. Results from 29 general practices. AB - AIMS: The aims of this study were to estimate the costs incurred by Dutch patients with Type 2 diabetes, examine which patient and/or treatment characteristics are associated with costs, and estimate the medical and non medical costs of patients with Type 2 diabetes in The Netherlands. METHODS: Twenty-nine Dutch general practitioners provided information on all Type 2 diabetes patients in their practice (n = 1371), information on demography, clinical characteristics, treatment type, the presence of complications and the type and amount of medical consumption during the previous 6 months. Medical costs were analysed using multivariate linear regression. Estimates of costs seen in The Netherlands were based on these results plus information from other sources regarding costs of end-stage renal disease, appliances, travel and productivity loss. RESULTS: Although only 9% of patients were hospitalized within the previous 6 months, hospitalization costs represented one-third of the medical costs, drug costs 40% and ambulatory costs 26%. Patients using insulin, patients with macrovascular complications only or in combination with microvascular complications incurred higher medical costs than other patients. Age and hyperlipidaemia were also positively related to medical costs. When these results were combined with other data sources, we estimated that patients with Type 2 diabetes are responsible for pound365 500 000 (1 271 000 000 guilders) or 3.4% of the relevant parts of health care costs in 1998. The non-medical costs (travel costs, productivity costs) are limited: 52 500 000 (183 000 000 guilders). CONCLUSIONS: Independent determinants of the medical costs of Type 2 diabetes in The Netherlands include age, complications, insulin use and hyperlipidaemia. PMID- 11918629 TI - 19th Anglo-Danish-Dutch Diabetes Group meeting. PMID- 11918628 TI - Age-related increase in haemoglobin A1c and fasting plasma glucose is accompanied by a decrease in beta cell function without change in insulin sensitivity: evidence from a cross-sectional study of hospital personnel. AB - AIMS: To examine the influence of age on glucose homeostasis in a population of healthy, non-diabetic hospital personnel. METHODS: One hundred and twenty female and 71 male non-diabetic individuals (fasting plasma glucose < 7.0 mmol/l) were fasted overnight prior to blood sampling. Glycated haemoglobin (HbA1c), fasting plasma glucose (FPG) and fasting plasma insulin (FPI) were measured using a BioRad Diamat automated HPLC, a Hitachi 747 analyser and a sensitive in-house radioimmunoassay, respectively. Mathematical modelling of the fasting glucose and insulin pairs (homeostasis model assessment (HOMA)) generated indices of pancreatic beta cell function, HOMA-B and tissue insulin sensitivity HOMA-S. RESULTS: Spearman rank correlation analysis showed that in the whole group there was a significant negative correlation between age and HOMA-B (rs = -0.218, P = 0.0022) and a significant positive correlation between age and both HbA1c (rs = 0.307, P = 0.0001) and FPG (rs = 0.26, P = 0.0003). There was no correlation between age and either FPI (rs = -0.08, P = 0.266) or HOMA-S (rs = 0.024, P = 0.75). Analysis by gender showed the above associations to be present in the females (rs = -0.243, P = 0.0076; rs = 0.304, P = 0.0007; rs = 0.32, P = 0.0004 for age vs. HOMA-B, HbA1c, and FPG, respectively). Again there was no correlation of age with FPI or insulin sensitivity. In the males there was a significant correlation of HbA1c with age (rs = 0.35, P = 0.002), but no significant correlation of age with any of the other parameters. CONCLUSIONS: Glycaemic control deteriorates with age in healthy, non-diabetic individuals. Age-related rises in FPG and haemoglobin A1c result from a small but steady decline in pancreatic beta cell function. PMID- 11918630 TI - Silent hypoglycaemia at the diabetic clinic. PMID- 11918631 TI - Analysis of a CD40 ligand dinucleotide microsatellite in multiple sclerosis. AB - In recent years, numerous reports have described the diverse roles of the CD40 CD40 ligand receptor-ligand pair. The interaction of these two cell-surface molecules regulates both humoral and cell-mediated immune functions. Because the CD40 ligand is known to be highly expressed on the peripheral blood mononuclear cells (PBMC) of multiple sclerosis (MS) patients, and because activated helper T cells expressing CD40 ligand have been found in the brain sections of MS patients, but not in those of normal controls, the protein is believed to be involved in MS development. We studied the influence of a polymorphic dinucleotide-repeat marker located in the 3' untranslated region of the X-linked gene encoding CD40 ligand (CD40LG) on susceptibility to and disease severity in MS. From a total cohort of 771 Nordic definite-MS patients, the most (n = 92) and least (n = 90) disabled octiles, as well as random samples of intermediately disabled males (n = 119) and females (n = 121), were genotyped; 135 ethnically matched healthy subjects were used as controls. In addition, the effect of the polymorphism on CD40 ligand mRNA expression was assessed using PBMC from 54 MS patients and 22 controls. The phenotype frequencies for the CD40LG marker did not differ significantly between gender-conditioned intermediate-MS subgroups and controls, or between gender-conditioned disability octiles. Nor did the polymorphism appear to exert any significant effect on mRNA expression in either patients or controls. PMID- 11918632 TI - cDNA cloning of the polymeric immunoglobulin receptor of the marsupial Macropus eugenii (tammar wallaby). AB - cDNA encoding a marsupial polymeric immunoglobulin receptor (pIgR) was isolated from Macropus eugenii (tammar wallaby) mammary lymph node primarily by reverse transcriptase coupled polymerase chain reaction (RT-PCR) and rapid amplification of cDNA ends (RACE) PCR. This resulted in a 5' truncated clone and, in order to obtain the full-length sequence, genomic walking PCR was utilized. The complete sequence consists of 2696 bp of cDNA and encodes a predicted polypeptide of 732 amino acids. The wallaby sequence is highly conserved in relation to the only other reported marsupial pIgR sequence, that of Trichosurus vulpecula (brushtail possum), having a nucleotide identity of 86.7% and a deduced amino acid identity of 79.9%. The wallaby nucleotide sequence also has a moderate degree of similarity with the pIgR sequences of eutherian mammals, being most similar to that of the rat, with an identity of 63.1%. At the amino acid level, in comparison to eutherian sequences, the wallaby pIgR is most similar to that of humans with an identity of 52.6%. pIgR phylogenetic trees were constructed for tammar wallaby, brushtail possum and several eutherian mammal cDNA and deduced amino acid sequences. In both DNA and protein analyses, the eutherian sequences formed a sister clade to the exclusion of the marsupial sequences, in agreement with the current view of mammalian evolution. PMID- 11918633 TI - Molecular cloning of four lambda light chain cDNAs from the Australian brushtail possum Trichosurus vulpecula. AB - A brushtail possum mesenteric lymph node cDNA library was screened with a grey short-tail opossum Clambda probe and four immunoglobulin lambda cDNAs were isolated. Two of the isolated clones (L5 and L10) contained identical framework 4 regions and constant regions (but different variable regions), suggesting that the possum lambda locus is organized as multiple J-C pairs--a feature seen in the opossum and placental mammals. The cloning of the lambda light chain cDNAs signifies the completion of the basic molecular characterization of the brushtail possum immunoglobulin repertoire. The availability of this sequence data will allow extensive analysis of the immune response of the brushtail possum at the molecular level, as well as the development of specific immunological reagents for detection of immunoglobulin molecules at the protein level. PMID- 11918634 TI - Molecular variation of HLA class I genes in the Corsican population: approach to its origin. AB - Three human leucocyte antigen (HLA) class I loci (HLA-A, -B and -Cw) were typed for the first time at the DNA level in the Corsican population. This analysis was performed on 100 individuals of Corsican origin living in central Corsica (46 individuals) and in south-western Corsica (54 individuals). The genetic structure of these two subpopulations was analysed on the basis of the molecular polymorphisms of the HLA-A, -B and -Cw genes. A phylogenetic and a haplotypic analysis were performed. The genotypic analysis did not detect genetic differentiation. Examination of the allelic and haplotypic frequencies did, however, reveal some significant differences between the two zones. Similarities with the Sardinian population were found, and were particularly evident in the south-western sample. However, Corsica has probably been subject to greater West European influence, which can be seen in the Corte sample (Haute Corse). PMID- 11918635 TI - A PCR-RFLP typing method for adhesion molecule gene polymorphisms and allele frequencies in a normal UK population. AB - We report simple and reproducible PCR-RFLP typing methods for the polymorphisms in the ICAM-1, E-selectin and PECAM-1 genes. The genotype and allele frequencies detected in a normal UK population did not deviate significantly from the Hardy Weinberg equilibrium; neither did they differ from frequencies previously reported using SSP or SSCP methods. PMID- 11918636 TI - Nomenclature for factors of the HLA system, update October 2001. PMID- 11918637 TI - Nomenclature for factors of the HLA system, update November 2001. PMID- 11918638 TI - Nomenclature for factors of the HLA system, update December 2001. PMID- 11918639 TI - Abstracts of the 16th European Histocompatibility Conference. March 19-22, 2002. Strasbourg, France. PMID- 11918642 TI - The modern management of adult spasticity: an evidence-based approach. Proceedings of a symposium. Madrid, Spain. October, 2000. PMID- 11918643 TI - The pathophysiology of spasticity. AB - Spasticity is only one of several components of the upper motor neurone (UMN) syndrome, known collectively as the 'positive' phenomena, that are characterized by muscle overactivity. Other components include tendon hyper-reflexia, clonus, the clasp-knife phenomenon, flexor and extensor spasms, a Babinski sign, and spastic dystonia. Spasticity is a form of hypertonia due to hyperexcitable tonic stretch reflexes. It is distinguished from rigidity by its dependence upon the speed of the muscle stretch and by the presence of other positive UMN signs. Hyperactive spinal reflexes mediate most of these positive phenomena, while others are due to disordered control of voluntary movement or abnormal efferent drive. An UMN lesion disturbs the balance of supraspinal inhibitory and excitatory inputs, producing a state of net disinhibition of the spinal reflexes. These include proprioceptive (stretch) and nociceptive (flexor withdrawal and extensor) reflexes. The clinical syndrome resulting from an UMN lesion depends more upon its location and extent, and the time since it occurred, than on the pathology of the lesion. However, the change in spinal reflex excitability cannot simply be due to an imbalance in supraspinal control. The delayed onset after the lesion and the frequent reduction in reflex excitability over time, suggests plasticity in the central nervous system. Knowledge of the electrophysiology and neurochemistry of spinal reflexes, together with the action of antispasticity drugs, helps us to understand the pathophysiology of spasticity. PMID- 11918644 TI - Outcome measures of spasticity. AB - Spasticity is characterized by a velocity-dependent increase in muscle resistance, in response to a passive stretch. Whilst clinical scales probably represent the most common approach to its measurement, these are limited by poor reliability and difficulties with some definitions. However, recently, a simple system has been used to provide the clinician with a measure of the force applied during the Ashworth Test. The Wartenberg Pendulum Test has been devised for measuring spasticity at the knee, but has been shown to be unsuitable for measuring more severe spasticity. Powered systems have been used in research studies, but are rarely considered suitable for routine clinical use. However, one method of interest, using a low inertia torque motor to measuring stiffness at the wrist, has been shown to provide rapid measurements which correspond to the degree of spasticity defined by other scales. Clinical gait analysis may have an important role to play in assessing spasticity - since there are clear associations between lower limb spasticity and gait problems - but it does not provide a true measure of the condition. In summary, while scales remain the most common method of measuring spasticity, there is considerable potential in instrumented techniques that can provide greater reliability and precision of measurement. PMID- 11918645 TI - Physical therapy in spasticity. AB - Physiotherapists are part of the rehabilitation team involved in the management of adult spasticity. Physical therapy is one part of the armoury in the fight against this disabling symptom of the upper motor neurone syndrome. Identifying the physiological changes brought about by physical therapy or physiotherapy, is difficult. As with many interventions in rehabilitation, this area is poorly studied and, until recently, there was little or no evidence for its effectiveness. The aim of this presentation is to identify key components of a physiotherapy approach and outline specific techniques. The key components considered are: education of the patient and their carers; the 'intervention cycle' - involving accurate assessment, careful measurement, intervention and evaluation; accurate goal setting and a staged stepwise approach over prolonged periods. The specific techniques used include treatments targeted at: muscle length changes, muscle strengthening and functional performance. Broadly these techniques can be divided into biomechanical, cognitive and neurophysiological, each being interdependent on the others. Physiotherapy combined with the other available treatments should meet the challenge arising from adult spasticity. PMID- 11918646 TI - Gait training in hemiplegia. AB - Restoration of gait is a major goal in neurological rehabilitation. Before starting therapy, a comprehensive assessment is necessary to evaluate the deficits and remaining functions. A wide variety of therapeutic procedures are available and have to be adapted to the individual situation - different concepts of physiotherapy stress different features like: force exercise, reduction of spasticity, gait symmetry, utilization of equilibrium reflexes, stepping automation, endurance training, repetition of rhythmic movements, etc. The spectrum of available therapies was recently widened by treadmill training, with partial body-weight support, locomotor pharmacotherapy, selective reduction of spasticity by botulinum toxin injections, and by musical biofeedback, which have each proved to be successful in the restoration of gait pattern. Treadmill training based on partial body weight support, combined with enforced stepping movements has proved to be successful in the restoration of gait pattern. A common problem in hemiparetic gait, is the spastic inversion of the foot. If spasticity is not severe, an ankle-foot orthosis (AFO) is the appropriate technical aid. In other cases, botulinum toxin injection into spastic leg muscles has been successfully used to improve gait functions. In hemiparetic stroke patients, auditory (musical) rhythm, as a peripheral pacing signal, resulted in a significant increase in weight-bearing stance time on the paretic side. In addition, there was an improved stride symmetry with rhythmic cueing and a normalizations of gait pattern. These methods directed to gait improvement should be combined and adapted to the individual patient's needs, in order to obtain the best results. PMID- 11918647 TI - The medical management of spasticity. AB - When spasticity produces a clinical disability by interfering with posture, motor capacity, nursing or daily living activities, medical treatment is recommended. It is mainly indicated when the muscle overactivity is diffusely distributed and should be implemented early, to prevent permanent musculoskeletal deformities or contractures. A pharmacological approach relies on the use of drugs which modulate neurotransmitters acting at the cortico-spinal level (GABA, glycine, glutamate, noradrenaline, serotonin). The aim of this treatment is to decrease spinal reflex excitability by reducing the release of excitatory neurotransmitters, or by potentiating the activity of inhibitory inputs. Evaluation of the efficacy of these drugs is determined by the therapeutic objectives which may be biomechanical, or functional. Diazepam increases presynaptic inhibition by stimulating GABA(A) receptors in the brainstem and spinal cord. In double-blind studies of patients with spinal cord lesions, antispastic efficacy has been shown, but side-effects are common. Baclofen stimulates GABA(B) receptors inducing a suppression of excitatory neurotransmitter release. Antispastic efficacy is sufficiently documented, but no definite effects on ambulation or activities of daily living have been proved. Tizanidine has an alpha2-agonist activity (at spinal and supraspinal level) and decreases the presynaptic activity of excitatory interneurones. The main clinical effects are a reduction in tonic stretch, polysynaptic reflexes, and co contraction, with fewer side-effects but no definite functional change. The efficacy of several other antispastic drugs is documented in a few controlled studies, but the majority of information arises from open trials or anecdotal observations. PMID- 11918648 TI - The surgical management of spasticity. AB - Neurosurgery is only considered for severe spasticity following the failure of noninvasive management (adequate medical and physical therapy). The patients are carefully selected, based on rigorous multidisciplinary clinical assessment. In this we evaluate the contribution of the spasticity to the disability and any residual voluntary motor function. The goals for each patient are: (a) improvement of function and autonomy; (b) control of pain; and (c) prevention of orthopaedic disorders. To achieve these objectives, the surgical procedure must be selective and reduce the excessive hypertonia without suppressing useful muscle tone and limb functions. The surgical procedures are: (1) Classical neuro ablative techniques (peripheral neurotomies, dorsal rhizotomies) and their modern modifications using microsurgery and intra-operative neural stimulation (dorsal root entry zone: DREZotomy). These techniques are destructive and irreversible, with the reduced muscle tone reflecting the nerve topography. It is mainly indicated when patients have localized spasticity without useful mobility. (2) Conservative techniques based on a neurophysiological control mechanism. These procedures are totally reversible. The methods involve chronic neurostimulation of the spinal cord or the cerebellum. There are only a few patients for whom this is indicated. Conversely, chronic intrathecal administration of baclofen, using an implantable pump, is well established in the treatment of diffuse spasticity of spinal origin. From reports in the literature, we critically review the respective indications in terms of function, clinical progression and the topographic extent of the spasticity. PMID- 11918649 TI - Botulinum toxin A (BoNT-A) for spasticity in adults. What is the evidence? AB - There are now many reports from open, uncontrolled studies which suggest that botulinum toxin A (BoNT-A) is valuable in treating spasticity. Evidence of its benefit is also gradually accumulating from randomized controlled trials (RCTs). In this presentation I will discuss the reasons why RCT evidence is being generated, and describe the findings currently available, including preliminary results from as yet unpublished trials. RCT data have been reported for leg and arm spasticity in a variety of diseases, but predominantly in stroke and multiple sclerosis patients. In most RCTs, the effects of BoNT-A are compared with placebo over a single injection cycle. The outcomes are generally positive and support the use of BoNT-A. However, data from RCTs are less convincing than those from open studies for a variety of technical reasons. These especially reflect the difficulties of finding good outcome measures for such a heterogeneous array of patients. There is good evidence that BoNT-A has clinical benefit in treating the mechanical effects of spasticity. In order to further clarify its usefulness, future research should address the strategies of short- and longer-term use of BoNT-A, and the unresolved technical issues of how to get the best out of this new treatment. PMID- 11918650 TI - A summary of spasticity management--a treatment algorithm. AB - The muscle overactivity seen in spasticity results in limb stiffness and muscle spasm, to which there is both a neurogenic and a biomechanical component. Spasticity does not always cause harm and can assist in the rehabilitation process enabling a patient to stand when their limb weakness would not otherwise allow it. When it does cause harm, however, treatment is required. This aims to (i) prevent provocative factors (ii) treat muscle overactivity; and (iii) prevent complications. Untreated, limb contracture, pain and other complications occur and early management can be most effective. Treatment is essentially physical, but, when this is inadequate, pharmacological intervention may be required. A strategy has been devised which shows that the first choice pharmacological treatment of focal spasticity is botulinum toxin. Over the past decade, the choice of treatment has become more ambitious with the establishment of new technologies. Good management now depends on an understanding of their role and application in relation to the needs of individual patients. To this end, a treatment algorithm which covers the salient facts in patient assessment and gives the indications for the range of available treatments, is the best approach. The indications and limitations of the available treatments are discussed, along with their place in the overall management of the patients. The evidence base for much of what is done is not strong and this summary examines the activities of proven value and of consensus view. PMID- 11918652 TI - Morphological and pharmacological evidence for the role of peripheral prostaglandins in the pathogenesis of neuropathic pain. AB - Inflammatory mediators produced in the injured nerve have been proposed as contributing factors in the development of neuropathic pain. Prostaglandins (PGs) are probably included in these important inflammatory mediators. In the present study, 2 and 4 weeks following partial sciatic nerve ligation (PSNL), we observed a dramatic increase in the prostaglandin synthesizing enzyme cyclooxygenase (COX)2-immunoreactive (IR) cell profiles in the injury site and adjacent region. Some of these COX2-IR cells were identified as macrophages because they coexpressed ED1. None of these COX2-IR cell profiles coexpressed the Schwann cell marker S100. In the contralateral sciatic nerve and sciatic nerve from normal rats, we failed to observe any of these COX-IR cell profiles. We also observed COX1-IR cell profiles (presumably Langerhans cells) in the epidermis of the footpad of both normal and PSNL rats. Interestingly, a greater number of COX1-IR cell profiles were observed in the epidermis of the ipsilateral footpad of PSNL rats. Local injection of ketorolac, a nonselective COX inhibitor, into the ipsilateral plantar side or into the injury site of the sciatic nerve, effectively reversed the tactile allodynia induced by PSNL for > 5 days. Intraperitoneal or intramuscular injection of ketorolac had a similar but shorter antiallodynic effect. Intraplantar or peri-neural injection of ketorolac dramatically suppressed the PSNL-induced increase in the phosphorylation of a transcription factor cAMP response element binding protein (CREB) in the ipsilateral dorsal horn of L4 and L5 spinal cord of PSNL rats. Intraplantar or peri-neural injection of ketorolac at the time of lesion did not prevent mechanical hypersensitivity but reduced it with a slow onset 3 weeks after lesion. Our data suggest that PSNL induces over-production of PGs in peripheral tissues and that PGs probably sensitize nociceptors and are involved in central plasticity and sensitization at the spinal cord level, thus contributing to the maintenance of tactile allodynia. PMID- 11918653 TI - Cerebral glucose utilization in transgenic mice overexpressing heat shock protein 70 is altered by dizocilpine. AB - Heat shock protein (HSP70), a member of the 70 kDa HSP superfamily, has been widely implicated in the cellular stress response to numerous insults. HSP70 may be a significant factor in cell survival following stresses such as cerebral ischaemia. The precise mechanisms by which HSP70 facilitates cell survival remain unclear. The aim of this study was to ascertain whether any differences in local cerebral glucose utilization (LCGU) existed between transgenic mice overexpressing HSP70 (HSP70 Tg) and wild- type littermate (WT) mice. LCGU was assessed using (14)C-2-deoxyglucose in HSP70 Tg and WT mice under basal conditions (intraperitoneal injection of saline) and during metabolic activation produced by NMDA receptor blockade (intraperitoneal injection of dizocilpine, 1 mg/kg). No significant alterations in LCGU were observed between saline injected HSP70 Tg and WT mice in any of the 35 brain regions analyzed. Dizocilpine injection produced significant heterogeneous alterations in LCGU in HSP70 Tg mice (24 of 35 brain regions) and in WT mice (22 of 35 brain regions) compared with saline injected mice. The distribution of altered LCGU produced by dizocilpine was similar in HSP70 Tg and WT mice. However in five brain regions, dizocilpine injected HSP70 Tg mice displayed significantly altered LCGU compared to dizocilpine injected WT mice (anterior thalamic nucleus +27%, dorsal CA1 stratum lacunosum molecularae +22%, dorsal CA1 stratum oriens + 14%, superior olivary body -26%, and the nucleus of the lateral lemniscus -16%). These data highlight that while overexpression of HSP70 transgene does not significantly alter LCGU in the basal state, mice overexpressing the HSP70 transgene respond differently to metabolic stress produced by NMDA receptor blockade in some important brain regions. PMID- 11918654 TI - Cooperative endocannabinoid production by neuronal depolarization and group I metabotropic glutamate receptor activation. AB - Endocannabinoids are retrograde messengers that are released from central neurons by depolarization-induced elevation of intracellular Ca2+ concentration [Ca2+]I or by activation of a group I metabotropic glutamate receptor (mGluR). We studied the interaction between these two pathways for endocannabinoid production in rat hippocampal neurons. We made a paired whole-cell recording from cultured hippocampal neurons with inhibitory synaptic connections. Activation of group I mGluRs, mainly mGluR5, by the specific agonist (RS)-3,5-dihydroxyphenylglycine (DHPG), suppressed inhibitory postsynaptic currents (IPSCs) in about half of the neuron pairs. A cannabinoid agonist, WIN55,212-2, suppressed IPSCs in all DHPG sensitive pairs but not in most of DHPG-insensitive pairs. The effects of both DHPG and WIN55,212-2 were abolished by the cannabinoid antagonists, AM281 and SR141716A, indicating that activation of group I mGluR releases endocannabinoids and suppress inhibitory neurotransmitter release through activation of presynaptic cannabinoid receptors. Depolarization of the postsynaptic neurons caused a transient suppression of IPSCs, a phemomenon termed depolarization induced suppression of inhibition (DSI) that was also abolished by cannabinoid antagonists. Importantly, DSI was enhanced significantly when group I mGluRs were activated simultaneously by DHPG. This enhancement was much more prominent than expected from the simple summation of depolarization-induced and group I mGluR induced endocannabinoid release. DHPG caused no change in depolarization-induced Ca2+ transients, indicating that the enhanced DSI by DHPG was not due to the augmentation of Ca2+ influx. Enhancement of DSI by DHPG was also observed in hippocampal slices. These results suggest that two pathways work in a cooperative manner to release endocannabinoids via a common intracellular cascade. PMID- 11918655 TI - Dynamics of nitric oxide during simulated ischaemia-reperfusion in rat striatal slices measured using an intrinsic biosensor, soluble guanylyl cyclase. AB - Nitric oxide (NO) may act as a toxin in several neuropathologies, including the brain damage resulting from cerebral ischaemia. Rat striatal slices were used to determine the mechanism of enhanced NO release following simulated ischaemia and, for estimating the NO concentrations, the activity of guanylyl cyclase served as a biosensor. Exposure of the slices for 10 min to an oxygen- and glucose-free medium caused a 70% fall in cGMP levels. On recovery, cGMP increased 2-fold above basal, where it remained for 40 min before declining. The pattern of changes matched those of cGMP or NO oxidation products measured during and after brain ischaemia in vivo. The increase observed during the recovery period was blocked by inhibition of NO synthase or NMDA receptors and was curtailed by tetrodotoxin, implying that it was caused by glutamate release leading to activation of the NMDA receptor-NO synthase pathway. Calibration of the cGMP levels against NO stimulated guanylyl cyclase yielded a basal NO concentration of 0.6 nm. The peak NO concentration achieved on recovery from simulated ischaemia was estimated as 0.8 nm. These values are compatible with the low micromolar concentrations of NO oxidation products (chiefly nitrate) found by microdialysis in vivo, providing the NO inactivation rate (forming nitrate) is accounted for. NO at a concentration around 1 nm is unlikely to be toxic to cells. However, if the NO inactivation mechanism were to fail (as it can) the NO production rate normally providing only subnanomolar NO could readily generate toxic (microM) NO concentrations. PMID- 11918656 TI - Spinal cord repair in neonatal rats: a correlation between axonal regeneration and functional recovery. AB - The present study aimed to analyse how anatomical regeneration contributes to functional recovery after experimental spinal cord repair. Thoracic spinal cord of neonatal rats was completely transected to make a gap and repaired by grafting a section of embryonic spinal cord. Six weeks after surgery, outcome of locomotor performance was assessed using an open field locomotor scale (BBB scale). Axonal regeneration across the repaired site was quantitatively assessed in the raphe, vestibular, and red nuclei and the sensorimotor cortex by a retrograde tracing method. The rats that had no labelled neurons in any of the supraspinal nuclei showed no hind-forelimb coordination. The rats that had labelled neurons in the brainstem nuclei but not in the sensorimotor cortex showed hind-forelimb coordination of varying grades depending on the amount of regeneration. The rats that had labelled neurons in all of the examined nuclei showed almost normal locomotion. In addition to a relationship between distribution of the labelled neurons and functional recovery, a positive correlation was observed between number of the labelled neurons in each of the supraspinal nuclei and locomotor performance of the rat. Thus the grade of restored function appeared to be regulated by distribution and number of fibres regenerated across the repaired site and into the target region. These results suggest that accurate reconstruction of neural connections is essential for significant functional recovery after spinal cord repair. PMID- 11918657 TI - Inhibition of spinal or hypoglossal motoneurons of the newborn rat by glycine or GABA. AB - The function of GABA or glycine during early postnatal development remains controversial as their action is reported as either excitatory or inhibitory. The present study addressed the question of the functional role of GABA or glycine on rat motoneurons shortly after birth. For this purpose, using in vitro preparations from immature rats (postnatal age, P0-P4 days), we recorded from lumbar spinal motoneurons and hypoglossal motoneurons. All data were obtained under current clamp conditions (recording with potassium methylsulphate containing electrodes) from cells at about -70 mV resting potential. On spinal motoneurons we used the glycinergic and GABAergic recurrent postsysnaptic potential (PSP) mediated by Renshaw cells to assess its impact on excitatory synaptic inputs from dorsal afferent fibres. Despite its depolarizing nature, the recurrent PSP consistently inhibited synaptic excitation of lumbar motoneurons. On hypoglossal motoneurons, exogenously applied GABA or glycine produced depolarization with decreased input resistance. This response was always associated with inhibition of cell firing induced by intracellular current pulses. Even when the membrane potential was repolarized to resting level in the presence of GABA or glycine, hypoglossal motoneurons failed to generate spikes. Conversely, similar depolarization produced by glutamate consistently facilitated spike firing. GABAergic and glycinergic synaptic potentials evoked by focal stimulation of the reticular formation inhibited firing and/or increased firing latency in the majority of hypoglossal motoneurons. These results indicate that, immediately after birth, rat motoneurons were inhibited by synaptically released or exogenously applied GABA or glycine. PMID- 11918658 TI - Synaptic depression improves coincidence detection in the nucleus laminaris in brainstem slices of the chick embryo. AB - Neurons in the nucleus laminaris detect the coincidence of binaural signals, and are the first neurons to calculate the interaural time difference for the sound source localization in birds. In this paper, we have studied contributions of synaptic depression to the coincidence detection in the nucleus laminaris in a slice preparation of the chick embryo (E16-18), using the whole-cell patch recording technique. Under voltage clamp, EPSCs decreased progressively in their amplitude during the course of tetanic stimuli. This synaptic depression was primarily ascribed to the reduction of transmitter release from the presynaptic terminal, because the depression was decreased by reducing transmitter release with 2.5 microm Cd2+ but was not affected by reducing desensitization of postsynaptic AMPA receptors with 20 microm cyclothiazide. Under current clamp, trains of 10 stimuli of 100 Hz were applied bilaterally with changing the time intervals systematically between both sides. Response window, defined as the time interval corresponding to the half-maximum firing probability, was narrowed during the course of the stimulus train, and this occurred in parallel with a decrease in the EPSP amplitude. In addition, the reduction of the EPSP amplitude due to 2.5 microm Cd2+ or 2 microm CNQX improved the accuracy of coincidence detection. These results indicate that the synaptic depression may improve the coincidence detection in the chick laminaris neurons. PMID- 11918659 TI - Neuroinflammation of the nigrostriatal pathway during progressive 6-OHDA dopamine degeneration in rats monitored by immunohistochemistry and PET imaging. AB - We investigated the microglial response to progressive dopamine neuron degeneration using in vivo positron emission tomography (PET) imaging and postmortem analyses in a Parkinson's disease (PD) rat model induced by unilateral (right side) intrastriatal administration of 6-hydroxydopamine (6-OHDA). Degeneration of the dopamine system was monitored by PET imaging of presynaptic dopamine transporters using a specific ligand (11)C-CFT (2beta-carbomethoxy-3beta (4-fluorophenyl) tropane). Binding of (11)C-CFT was markedly reduced in the striatum indicating dopaminergic degeneration. Parallel PET studies of (11)C PK11195 (1-(2-chlorophenyl)-N-methyl-N-(1-methylpropyl)-3 isoquinoline carboxamide) (specific ligand for activated microglia) showed increased binding in the striatum and substantia nigra indicative of a microglial response. Postmortem immunohistochemical analyses were performed with antibodies against CR3 for microglia/macrophage activation. Using a qualitative postmortem index for microglial activation we found an initially focal, then widespread microglial response at striatal and nigral levels at 4 weeks postlesion. These data support the hypothesis that inflammation is a significant component of progressive dopaminergic degeneration that can be monitored by PET imaging. PMID- 11918660 TI - Transistor array with an organotypic brain slice: field potential records and synaptic currents. AB - Linear transistor arrays on a silicon chip were used to map evoked extracellular field potentials in organotypic brain slices of rat hippocampus. The shape and amplitude of the transients were similar to those from records with micropipette electrodes. The spatial resolution was 21 and 4.6 microm. The sampled profile of the field potential showed a wide and shallow trough of transients in the stratum radiatum and a narrow but higher ridge in the stratum pyramidale. Due to the high resolution, the profile could be interpreted quantitatively in terms of synaptic currents. Transistor chips may become a novel tool for neurophysiological and pharmacological studies in brain slices. PMID- 11918661 TI - Differential expression of phospholipase D1 in the developing retina. AB - Expression patterns of phospholipase D1 (PLD1) in the developing rat retina were investigated using immunocytochemistry and Western blot analysis and compared with the expression patterns of glutamine synthetase. PLD1 immunoreactivity appeared first in a few neuroblasts in the middle of the mantle zone of the primitive retina by embryonic (E) day 13. PLD1-immunoreactive primitive ganglion cells were characterized in the ganglion cell layer by E17. Faint immunoreactivity at E17 profiled radially orientated cells and this pattern appeared up to postnatal (P) day 7. In the ganglion cell layer at P3, displaced amacrine cells and ganglion cells were classified. At P5, presumptive horizontal cells and amacrine cells were identified. By P7, a thin outermost layer of newly formed segments of the photoreceptor cells was also PLD1 immunoreactive. PLD1 immunoreactivity at P8 was limited to radial Muller cells and the outer segment layer of the photoreceptor cells, and the expression pattern was conserved to adulthood. Western blot analysis showed relatively high amounts of PLD1 protein at E17 and P3, a decrease at P7, and moderate amounts from P8 onward. Co expression of PLD1 with glutamine synthetase in the retina appeared first after birth in differentiating neurons and in Muller cells by P8; thereafter the pattern was maintained. The expression pattern of the PLD1 during development of the retina suggests that PLD1 plays important roles in glutamate-associated differentiation of both specific neurons and radial glial cells, and in glutamate mediated cellular signalling in Muller cells. PMID- 11918662 TI - Neuronal activation and corticotropin-releasing hormone expression in the brain of obese (fa/fa) and lean (fa/?) Zucker rats in response to refeeding. AB - The present study was conducted to investigate the pattern of neuronal activation and corticotropin-releasing hormone (CRH) expression in fed, food deprived and refed lean (Fa/?) and obese (fa/fa) Zucker rats. The pattern of neuronal activation was studied by measuring the expression of the immediate-early gene c fos. Expression of c-fos and CRH mRNA was determined by in situ hybridization histochemistry. In both lean and obese rats, one hour of refeeding led to a transient increase in c-fos mRNA levels which was detected in the paraventricular hypothalamic nucleus (PVH), the dorsomedial hypothalamic nucleus, the supraoptic nucleus, the paraventricular thalamic nucleus, the central nucleus of amygdala (CeA), the lateral and medial parabrachial nuclei, the nucleus of the solitary tract, and the area postrema. In addition, refeeding led to strong activation of the arginine-vasopressin neurons located in the magnocellular part of the PVH. Following 24 h of food deprivation, CRH expression in the parvocellular division of the PVH was significantly higher in obese rats compared to lean animals. During refeeding, PVH CRH mRNA levels in obese rats decreased to reach control values. The decrease in CRH expression in obese rats was accompanied by the alleviation of the hypercorticosteronemia that characterized obese Zucker rats. CRH mRNA levels in the central nucleus of the amygdala were significantly higher in lean rats than in obese animals, when the rats were fed ad libitum During food deprivation, CeA CRH mRNA levels decreased in lean rats and gradually returned to predeprivation values during refeeding. In refed obese rats, CeA levels of CRH mRNA were higher than those of ad libitum fed or food-deprived obese mutants. In the perifornical region of the lateral hypothalamic area (LHA), the expression of CRH mRNA rose significantly in response to refeeding in lean rats, but not in obese animals. Following the first hour of refeeding, the number of neurons expressing CRH mRNA in the LHA in lean rats almost doubled. The present results demonstrate that refeeding has a stimulating effect in obese Zucker rats in a pattern of activation similar to that seen in lean Fa/? rats. They also demonstrate differences in CRH expression between Fa/? and fa/fa rats after refeeding. The most apparent of these differences was seen in the lateral hypothalamus in which refeeding failed to up-regulate CRH expression in obese rats. PMID- 11918663 TI - Acute cholinergic rescue of synaptic plasticity in the neurodegenerating cortex of anti-nerve-growth-factor mice. AB - Deficits in cholinergic systems innervating cerebral cortex are associated with cognitive impairment during senescence and in age-related neurodegenerative pathologies. However, little is known about the role of cholinergic pathways in modulating cortical plasticity. Basal forebrain cholinergic neurons are a major target for nerve-growth factor (NGF). In order to investigate the relationship between cholinergic innervation and cortical synaptic plasticity, we exploited a transgenic mouse model in which the activity of NGF in the adult nervous system is neutralized by the expression of blocking antibodies to NGF itself (anti-NGF mice) [Ruberti, F. et al. (2000). J. Neurosci. 20, 2589-2601]. In 6-month-old anti-NGF mice, we show that the reduction in cholinergic innervation of the cortex is associated with different forms of synaptic plasticity impairment. A local, acute increase in the availability of acetylcholine rescues these synaptic plasticity deficits, thus indicating that a cholinergic system mediates the impairment of cortical plasticity at this early stage of the neurodegenerative process triggered by NGF neutralization. Our results represent an important step in unveiling the pivotal role of cholinergic transmission in modulating adult cortical plasticity. PMID- 11918665 TI - Sensitization to the neuroendocrine, central monoamine and behavioural effects of murine tumor necrosis factor-alpha: peripheral and central mechanisms. AB - Systemic administration of murine tumour necrosis factor-alpha (mTNF-alpha; 0.1 2.0 microg, i.p.) dose-dependently increased plasma corticosterone and augmented monoamine utilization within the paraventricular nucleus of the hypothalamus (PVN), locus coeruleus, medial prefrontal cortex (PFC), central and medial amygdala. A time-dependent sensitization was induced in mice, wherein reexposure to mTNF-alpha 28 days (but not 1 day) following the initial cytokine treatment provoked marked signs of illness (diminished activity, ptosis, piloerection) and increased plasma corticosterone levels. Serotonin (5-HT) activity was augmented upon mTNF-alpha reexposure at the 1- or 28-day intervals in the PFC and medial amygdala, respectively. Intracerebroventricular (i.c.v.; 1-500 ng) mTNF-alpha did not promote illness, but modestly increased plasma corticosterone levels. Neither the illness nor the corticosterone changes were subject to a sensitization upon i.c.v. cytokine reexposure. Acute i.c.v. mTNF-alpha increased norepinephrine (NE), 5-HT and dopamine (DA) activity within the PVN and median eminence/arcuate nucleus complex (ME/ARC), and NE utilization within the central amygdala. Subsequent i.c.v. mTNF-alpha further enhanced the hypothalamic monoamine variations. Finally, systemic (i.p.) mTNF-alpha pretreatment did not proactively influence sickness or corticosterone responses upon later i.c.v. cytokine challenge, but augmented locus coeruleus NE activity and 5-HT and DA utilization within the ME/ARC. It is suggested that the sensitization with respect to sickness and corticosterone activity in response to mTNF-alpha reflect the involvement of peripheral mechanisms. Moreover, it appears that mTNF-alpha promotes central neurochemical plasticity through independent central and peripheral mechanisms. PMID- 11918664 TI - Forced swimming evokes a biphasic response in CREB phosphorylation in extrahypothalamic limbic and neocortical brain structures in the rat. AB - The transcription factor cAMP response element-binding protein (CREB) plays a critical role in plasticity processes underlying learning and memory. We investigated the phosphorylation of CREB in rat brain after forced swimming, a stressor known to impact on higher limbic and neocortical brain areas. As shown by immunohistochemistry, forced swimming increased phosphorylated CREB (P-CREB) levels in the dentate gyrus, all neocortical areas, the medial, lateral and basolateral nuclei of the amygdala, cerebellum but not in the hypothalamic paraventricular nucleus. Distinct differences in the P-CREB pattern were observed in the deeper vs. superficial layers of the neocortex. The response in P-CREB was stressor type-specific because exposure to either ether or a cold environment was ineffective. The forced swimming-induced changes in P-CREB levels showed a biphasic time-course: an early peak detected at 15 min was followed by a marked drop at 60 min; a second rise starting after 1-2 h, reached maximal values between 6 and 8 h, and remained elevated for at least 48 h. Examination of the neuroanatomical induction pattern of the CRE-inducible immediate early gene product c-fos revealed that it was only partly overlapping with that of P-CREB. Western analyses showed that only the 43-kDa CREB protein (an enhancer of CRE containing promotors) was phosphorylated after forced swimming, while other members of the CREB/ATF family (CREM, ATF-1 and ATF-2) remained unaffected. The NF-kappaB pathway was not activated, indicating that forced swimming does not unspecifically evoke transcription factor activation. Thus, in contrast to physical stressors, such as ether or cold exposure, forced swimming, a stressor with a strong psychological component, elicits the recruitment of the CREB pathway in a widespread manner in the limbic system and neocortex; brain regions known to be implicated in various forms of (stress-related) learning and memory. PMID- 11918667 TI - Memory traces for inflectional affixes as shown by mismatch negativity. AB - Mismatch negativity (MMN), an index of experience-dependent memory traces, was used to investigate the processing of grammatical affixes in the human brain. The MMN was elicited by either a verb stem or an inflected verb form, phonetic contrasts being identical in both conditions. The topography of the mismatch responses showed clear left-hemispheric laterality in both conditions. However, the MMN to the inflected form occurred later than that for the stem. Furthermore, the inflected stimulus produced MMN maximal in centroparietal sites, whereas stem elicited MMN was more profound at more frontal sites. We suggest that these features of the MMN to inflected form indicate delayed activation of left lateralized perisylvian cell assemblies that function as cortical memory traces of inflectional affixes. PMID- 11918666 TI - Superposition of horseshoe-like periodicity and linear tonotopic maps in auditory cortex of the Mongolian gerbil. AB - The segregation of an individual sound from a mixture of concurrent sounds, the so-called cocktail-party phenomenon, is a fundamental and largely unexplained capability of the auditory system. Speaker recognition involves grouping of the various spectral (frequency) components of an individual's voice and segregating them from other competing voices. The important parameter for grouping may be the periodicity of sound waves because the spectral components of a given voice have one periodicity, viz. fundamental frequency, as their common denominator. To determine the relationship between the representations of spectral content and periodicity in the primary auditory cortex (AI), we used optical recording of intrinsic signals and electrophysiological mapping in Mongolian gerbils (Meriones unguiculatus). We found that periodicity maps as an almost circular gradient superimposed on the linear tonotopic gradient in the low frequency part of AI. This geometry of the periodicity map may imply competitive signal processing in support of the theory of "winner-takes-all". PMID- 11918668 TI - The interaction between sigmaS, the stationary phase sigma factor, and the core enzyme of Escherichia coli RNA polymerase. AB - BACKGROUND: The RNA polymerase holoenzyme of Escherichia coli is composed of a core enzyme (subunit structure alpha2betabeta') associated with one of the sigma subunits, required for promoter recognition. Different sigma factors compete for core binding. Among the seven sigma factors present in E. coli, sigma70 controls gene transcription during the exponential phase, whereas sigmaS regulates the transcription of genes in the stationary phase or in response to different stresses. Using labelled sigmaS and sigma70, we compared the affinities of both sigma factors for core binding and investigated the structural changes in the different subunits involved in the formation of the holoenzymes. RESULTS: Using native polyacrylamide gel electrophoresis, we demonstrate that sigmaS binds to the core enzyme with fivefold reduced affinity compared to sigma70. Using iron chelate protein footprinting, we show that the core enzyme significantly reduces polypeptide backbone solvent accessibility in regions 1.1, 2.5, 3.1 and 3.2 of sigmaS, while increasing the accessibility in region 4.1 of sigmaS. We have also analysed the positioning of sigmaS on the holoenzyme by the proximity-dependent protein cleavage method using sigmaS derivatives in which FeBABE was tethered to single cysteine residues at nine different positions. Protein cutting patterns are observed on the beta and beta' subunits, but not alpha. Regions 2.5, 3.1 and 3.2 of sigmaS are close to both beta and beta' subunits, in agreement with iron chelate protein footprinting data. CONCLUSIONS: A comparison between these results using sigmaS and previous data from sigma70 indicates similar contact patterns on the core subunits and similar characteristic changes associated with holoenzyme formation, despite striking differences in the accessibility of regions 4.1 and 4.2. PMID- 11918669 TI - A GyrB-GyrA fusion protein expressed in yeast cells is able to remove DNA supercoils but cannot substitute eukaryotic topoisomerase II. AB - BACKGROUND: Type II topoisomerases are a highly conserved class of enzymes which transport one double-stranded DNA segment through a transient break in another. Whereas the eukaryotic enzymes are homodimers of a single polypeptide, their bacterial homologues are homodimers of two independently coded protein subunits. Unlike eukaryotic topoisomerase II and bacterial topoisomerase IV, DNA gyrase is a bacterial type II topoisomerase which specializes in intramolecular DNA transport. RESULTS: We have fused the Escherichia coli coding sequences for the proteins GyrB and GyrA, which comprise DNA gyrase. This fusion was expressed in yeast cells and yielded the expected full-length protein product. When it was expressed in Deltatop1- top2-4 yeast cells, the fusion protein compensated their slow growth and reverted their elevated chromosomal excision of ribosomal genes. Furthermore, it removed DNA positive supercoils. The fusion protein, however, was unable to complement the temperature-dependent lethality of top2-4 cells. CONCLUSION: Fusion of the E. coli GyrB and GyrA proteins leads to a catalytically active topoisomerase which compensates several phenotypic traits attributed to unconstrained DNA supercoiling in topoisomerase-deficient cells. However, since the fusion protein cannot substitute for topoisomerase II, it may be efficient in intramolecular but not intermolecular DNA passage, resembling the catalytic properties of DNA gyrase. PMID- 11918671 TI - Level of the RNA polymerase II in the fission yeast stays constant but phosphorylation of its carboxyl terminal domain varies depending on the phase and rate of cell growth. AB - BACKGROUND: The RNA polymerase II of the fission yeast Schizosaccharomyces pombe consists of 12 Rpb subunits, of which four (Rpb1, Rpb2, Rpb3 and Rpb11) form the assembly and catalytic core and five (Rpb5, Rpb6, Rpb8, Rpb10 and Rpb12) are shared among RNA polymerases I, II and III. The intracellular levels of three RNA polymerase forms should be interrelated, but the control of RNA polymerase formation remains mostly unknown. RESULTS: To reveal the physiological role and the synthesis control of each Rpb subunit, the intracellular levels of the Rpb proteins were examined in S. pombe growing at various phases under various conditions. Results indicate that the intracellular concentrations of the Rpb proteins stay constant at levels characteristic of the rate and phase of cell growth, and the relative level between the 12 subunits also remains constant, together implying that the intracellular concentration of RNA polymerase II stays constant, as in the case of prokaryotes. As an attempt to gain insights into the activity control of RNA polymerase II, we also analysed the phosphorylation level of the carboxyl-terminal domain (CTD) of the largest subunit Rpb1. Phosphorylated forms of Tyr1 and Thr4 within 29 repeats of the YSPTSPS heptapeptide were detected in both slow-migrating IIo and fast-migrating IIa forms of Rpb1 on SDS PAGE (polyacrylamide gel electrophoresis). However, phosphorylated Ser2 and Ser5 were identified only in the IIo form, indicating that Ser phosphorylation contributes to the conformational change in CTD. The phosphorylation levels of Ser, Thr and Tyr all vary depending on the cell culture conditions. CONCLUSION: The intracellular level of RNA polymerase II stays constant, but the amount engaged in transcription cycle varies depending on the culture conditions, as estimated from the sites and levels of phosphorylation of Rpb1 CTD. PMID- 11918670 TI - Identification and characterization of tRNA (Gm18) methyltransferase from Thermus thermophilus HB8: domain structure and conserved amino acid sequence motifs. AB - BACKGROUND: Transfer RNAs from an extreme thermophile, Thermus thermophilus, commonly possess 2'-O-methylguanosine at position 18 (Gm18) in the D-loop. This modification is post-transcriptionally introduced by tRNA (Gm18) methyltransferase. RESULTS: Partial amino acid sequence data were obtained from purified T. thermophilus tRNA (Gm18) methyltransferase by peptide sequencing and mass spectrometry. The sequence data were used to screen the T. thermophilus genome database currently in progress, resulting in the identification of the corresponding gene. Purified recombinant enzyme showed a strict specificity for methylation at the 2'-OH of G18 in tRNA. Sequence alignment with other known or putative methyltransferases elucidates that tRNA (Gm18) methyltransferases have specific conserved region as well as three consensus motifs found in RNA ribose 2'-O-methyltransferases. The enzyme truncated at its N and C termini by limited tryptic digestion still retained binding activity for S-adenosyl-l-homocysteine, but lost the catalytic activity. CONCLUSION: This is the first report describing the identification of a methyltransferase gene of the trmH family through the analysis of a purified protein. Further, our results indicate that a restricted region(s) in the terminal amino acid residues of T. thermophilus tRNA (Gm18) methyltransferase are responsible for tRNA recognition and a main part of the enzyme is allocated for a catalytic core. PMID- 11918672 TI - ICRF-193, a catalytic inhibitor of DNA topoisomerase II, inhibits re-entry into the cell division cycle from quiescent state in mammalian cells. AB - BACKGROUND: To describe the requirement of DNA topoisomerase II (topo II) during transition from the quiescent state (G0 phase) to the cell division cycle in mammalian cells, we examined the influence of ICRF-193, a catalytic inhibitor of topo II, on re-entry into the cell division cycle of quiescent cells in response to appropriate growth stimuli. RESULTS: The re-entry into the S phase of cultured cell lines arrested at the quiescent (G0) phase by serum-starvation was sensitive to 10 microm ICRF-193. DNA syntheses induced by lipopolysaccharide in murine spleen cells or by release from contact-inhibition were also inhibited by ICRF 193. The cell lines with a high-level of resistance toward ICRF-193 due to a point mutation in the topo IIalpha gene entered into the S phase from quiescence in the presence of ICRF-193. The drug did not inhibit entry into the S phase in cultured cells released from arrest at the metaphase or G1 phase. CONCLUSION: There is an ICRF-193-sensitive step during re-entry of quiescent mammalian cells into the cell division cycle upon growth stimulation and the drug targets topo IIalpha during the process. PMID- 11918673 TI - Distribution of phosphorylated glial fibrillary acidic protein in the mouse central nervous system. AB - BACKGROUND: Glial fibrillary acidic protein (GFAP) is the principal component of intermediate filaments (IFs) in mature astrocytes in the central nervous system (CNS). Like other IF proteins, GFAP has multiple phosphorylation sites in the N terminal head domain. The distribution of phospho-GFAP in vivo has not been elucidated. RESULTS: We generated Gfap(hwt) knock-in mice, in which the coding region for the head domain of GFAP is replaced with the corresponding human sequence. In combination with a series of monoclonal antibodies (mAbs) reactive to human phospho-GFAP, we visualized the distribution of phospho-GFAP in vivo in mice. GFAP phosphorylated at Thr7, Ser8 and/or Ser13 increased postnatally in the CNS of these mice. Limited populations of GFAP-positive astrocytes were labelled with anti-phospho-GFAP mAbs in most brain areas, whereas almost all the astrocytes in the optic nerve and spinal cord were labelled. Astrocytes in the subventricular zone and rostral migratory stream preferentially contained phospho GFAP. In a cold injury model of the cerebral cortex, we detected phospho-GFAP in reactive astrocytes at 2-3 weeks after the injury. CONCLUSIONS: Phospho-GFAP provides a molecular marker indicating the heterogeneity of astrocytes, and Gfap(hwt) knock-in mice will aid in monitoring intracellular conditions of astrocytes, under various conditions. Our results suggest that the phosphorylation of GFAP plays a role in non-dividing astrocytes in vivo. PMID- 11918674 TI - Requirement of the IFN-alpha/beta-induced CXCR3 chemokine signalling for CD8+ T cell activation. AB - BACKGROUND: Activation of both CD4+ T and CD8+ T cells is triggered by the engagement of the T cell antigen receptor (TCR) with MHC/peptide complexes on antigen-presenting cells. This process also requires other molecular interactions, which transmit co-stimulatory signals to these T cells. To ensure an effective immune response, distinct T cell subsets may additionally employ unique mechanism(s) for efficient activation. RESULTS: We here show that mutant CD8+ T cells lacking the IFN-alpha/beta signalling components are hyporesponsive to antigen stimulation in vitro. We further show that IFN-alpha/beta-mediated signals are required for induction of the chemokines IP-10/I-TAC and their common receptor, CXCR3, and in turn provide evidence that CXCR3-mediated signals indeed function in the activation and proliferation of CD8+ T cells, particularly for the CD44low naive phenotype cells. CONCLUSION: The CXCR3 chemokine system is regulated by IFN-alpha/beta in CD8+ T cells, and it is critical for the efficient cell activation. The present study therefore reveals a novel role of the IFN alpha/beta-CXCR3 signalling cascade in CD8+ T cell activation. PMID- 11918675 TI - The FYVE domain in Smad anchor for receptor activation (SARA) is sufficient for localization of SARA in early endosomes and regulates TGF-beta/Smad signalling. AB - BACKGROUND: Transforming growth factor-beta (TGF-beta) initiates intracellular signalling by inducing the formation of a heteromeric complex between TGF-beta type I (TbetaR-I) and TGF-beta type II serine/threonine kinase receptors (TbetaR II). After the activation of TbetaR-I kinase by TbetaR-II kinase, specific receptor-regulated Smads (R-Smads) are phosphorylated by TbetaR-I kinase. Smad anchor for receptor activation (SARA), which contains a FYVE finger domain, regulates the subcellular localization of R-Smads and presents them to TbetaR-I. However, it is unclear where SARA is localized in the cell and which phospholipid(s) interacts with its FYVE domain. RESULTS: Wild-type SARA and the FYVE domain of SARA (FYVE(SARA)) reveal a punctate staining pattern and co localize with the early endosomal markers, early endosomal antigen-1 (EEA1) and hepatic growth factor-regulated tyrosine kinase substrate (Hrs). The ectopic expression of dominant negative rab5, a critical regulatory molecule in endosome function, redistributes SARA from punctate to a diffuse cytosolic staining pattern. A lipid binding assay demonstrated that the recombinant FYVE domain from SARA predominantly interacts with phosphatidylinositol 3-phosphate (PtdIns(3)P). Consistent with this result, wortmannin, a PI3 kinase inhibitor, resulted in both a redistribution of SARA from the endosomal compartment to the cytosol and the attenuation of both TGF-beta-induced R-Smad phosphorylation and transcriptional activation. Ectopic expression of the FYVE domain of SARA also induced the redistribution of wild-type SARA and inhibited TGF-beta as well as BMP/Smad induced transcriptional responses. CONCLUSION: The FYVE domain is sufficient and necessary for the early endosomal localization of SARA, probably through its interaction with PtdIns(3)P. Moreover, the localization of SARA in early endosomes is required for efficient TGF-beta/Smad signalling. PMID- 11918677 TI - Detection of tmRNA-mediated trans-translation products in Bacillus subtilis. AB - BACKGROUND: Bacterial tmRNA (10Sa RNA) is involved in a trans-translation reaction, which contributes to the degradation of incompletely synthesized peptides and the recycling of stalled ribosomes. To investigate the physiological roles of this reaction in Bacillus subtilis, we devised a system for detecting the proteins that are subject to in vivo trans-translation. RESULTS: The wild type tmRNA gene (ssrA) in the genome was replaced by a variant ssrA encoding a tag-peptide sequence containing six histidine residues (His-tag) and two aspartic acids at the C-terminus. The His-tagged proteins that accumulated in the cells without degradation were fractionated by Ni2+-NTA column and gel electrophoresis and were detected by Western blotting with an anti-His-tag antibody. The results showed that the trans-translation occurred more frequently at a high temperature (50 degrees C) than at a low temperature (37 degrees C). Two-dimensional (2D) gel electrophoresis of the products revealed many distinct spots, which represent specific target proteins for the trans-translation reaction. Furthermore, the 2D gel patterns of the products from cells cultured at high and low temperatures were apparently different. Several tagged proteins were identified by the N terminal amino acid sequences of the products. CONCLUSION: Trans-translation occurs more frequently at high temperature than at low temperature, and different proteins are tagged at different temperatures. PMID- 11918676 TI - Characterization, expression and complex formation of the murine Fanconi anaemia gene product Fancg. AB - BACKGROUND: Fanconi anaemia (FA) is an autosomal recessive chromosomal instability disorder. Six distinct FA disease genes have been identified, the products of which function in an integrated pathway that is thought to support a nuclear caretaker function. Comparison of FA gene characteristics in different species may help to unravel the molecular function of the FA pathway. RESULTS: We have cloned the murine homologue of the Fanconi anaemia complementation group G gene, FANCG/XRCC9. The murine Fancg protein shows an 83% similarity to the human protein sequence, and has a predicted molecular weight of 68.5 kDa. Expression of mouse Fancg in human FA-G lymphoblasts fully corrects their cross-linker hypersensitivity. At mRNA and protein levels we detected the co-expression of Fancg and Fanca in murine tissues. In addition, mouse Fancg and Fanca proteins co purify by immunoprecipitation. Upon transfection into Fanca-deficient mouse embryonic fibroblasts EGFP-Fancg chimeric protein was detectable in the nucleus. CONCLUSIONS: We identified a murine cDNA, Fancg, which cross-complements the cellular defect of human FA-G cells and thus represents a true homologue of human FANCG. Spleen, thymus and testis showed the highest Fancg expression levels. Although Fancg and Fanca are able to form a complex, this interaction is not required for Fancg to accumulate in the nuclear compartment. PMID- 11918678 TI - Publishers' Note. PMID- 11918679 TI - Rat macrophages: membrane glycoproteins in differentiation and function. AB - Macrophages (mphi) play a crucial role in the immune system. The rat offers unique advantages for studying the biology of mphi. Firstly, monoclonal antibodies (mAb) against many rat mphi surface glycoproteins have become available. These have not only demonstrated a considerable heterogeneity among mphi, but have also allowed the characterization of various mphi surface molecules in terms of structure, expression regulation and function. Furthermore, substantial numbers of rat mphi can be isolated from various sites (e.g. blood, peritoneal cavity, lung and other tissues), enabling proper molecular and functional analysis of these mphi populations. Finally, a number of (unique) experimental models for human diseases have been developed in the rat, making possible the evaluation of the involvement of mphi in pathogenesis. For this, a method for the selective elimination of mphi from various tissues in vivo has proven very useful. Here, we will review the contribution that the rat has made to understanding the immunobiology of mphi. In particular, we will discuss the surface (glyco)proteins on rat mphi in differentiation and function, and the involvement of mphi in rat models of disease. PMID- 11918680 TI - Genomic and funtional aspects of the rat MHC, the RT1 complex. AB - The major histocompatibility complex (MHC) plays a central role in controlling immune responsiveness, susceptibility to certain diseases and histo incompatibility. A physical map of the complete rat MHC, the RTI complex, is presented based on a PAC clonal contig (RT1n haplotype). Expression profiling of various tissues of different inbred strains has been carried out for genes of the RT1-C/E/M region, and different types of variability of expression are shown. As an example of single gene analysis, the RT1-linked heat shock-inducible heat shock genes Hsp70-1 and Hsp70-2 have been studied. It is demonstrated that their gene products are able to increase lysability of target cells by cytotoxic T lymphocytes. PMID- 11918681 TI - Rat models as tool to develop new immunotherapies. AB - Rat models (experimental autoimmune encephalomyelitis, EAE; myasthenia gravis, EAMG) have been developed that mimic certain aspects of the pathophysiology of multiple sclerosis (MS) and myasthenia gravis (MG) in humans. Rat EAE and EAMG are, therefore, widely used to evaluate immunotherapeutic strategies. Mucosal tolerance induced by oral or nasal administration of autoantigen effectively suppresses rat EAE and EAMG. Nasal administration of the cytokines interleukin (IL)-4, IL-10 or transforming growth factor (TGF)-beta1 also ameliorates clinical signs of rat EAE. However, neither mucosal tolerance nor cytokines affect clinical disease if administered during ongoing rat EAE or EAMG. Dendritic cells (DC) pulsed in vitro with autoantigen can mediate peripheral tolerance against rat EAE and EAMG, but do not affect ongoing EAE or EAMG. DC from healthy rats, modified in vitro by exposure to IFN-gamma or TGF-beta1, effectively suppress ongoing EAE and EAMG when given by the subcutaneous route. Most importantly, DC from EAMG rats, after being modified in vitro with IL-10, also inhibit ongoing rat EAMG. These results demonstrate that different strategies influencing T- and/or B-cell responses in rat EAE or EAMG are available, but that DC-based immunotherapies are the most promising, in order to ameliorate ongoing organ specific autoaggressive immunity Based on these observations, autologous DC, after being modified in vitro with cytokines, constitute a basis for new immunotherapeutic strategies in MS, MG and other diseases with autoimmune background. PMID- 11918682 TI - The athymic nude rat: an animal experimental model to reveal novel aspects of innate immune responses? AB - Athymic nude rats resemble nude mice in their lack of a normal thymus and functionally mature T cells. They have been useful in the study of mechanisms of tumor growth or graft rejection in immunocompromised hosts since they can accept major histocompatibility complex (MHC) mismatched organ allografts or xenografts for several months and because a number of tumor cell lines of human and rodent origin grow well in these rats. Injection of a few helper T (Th) cells from euthymic littermate rats partly restores the pool of mature T cells as well as full immunocompetence to reject organ allografts and has helped to reveal some of the cell interactions necessary for rejection to occur In contrast, immunologically naive athymic nude rats of certain strains, acutely reject allografts consisting of lymphocytes or bone marrow cells, which is due to the presence of alloreactive natural killer cells. These cells can recognize and kill MHC incompatible hematopoietic cells through the recognition of both mismatches within the classical (RT1.A) and nonclassical (RT1.C/E) MHC class I regions with a repertoire of inhibitory and activating killer lectin-like receptors (KLR) for MHC-I molecules, encoded by the Ly-49 portion of the rat natural killer cell gene complex (NKC). Some of these receptors have been identified and molecularly cloned and show similarities with NK receptors identified in the mouse. Other leukocytes in nude rats, such as dendritic cells, may also contribute to specific innate immune responses in the absence of mature T cells. Nude rats develop T like cells expressing CD3 and T-cell receptor (TCR) with increasing age. Though their phenotype in peripheral lymphatic tissues resembles that of normal T cells, consisting mainly of CD4+ or CD8+ cells, they lack alloreactivity in vivo and their TCR repertoire is more of an oligoclonal nature. Their contribution to allograft rejection in T-cell-reconstituted rats is therefore questionable, and their role in innate immune response in these rats still enigmatic. PMID- 11918683 TI - Neuroendocrine responses regulating susceptibility and resistance to autoimmune/inflammatory disease in inbred rat strains. AB - Rodent animal models of inflammatory and autoimmune disease have been important tools in the study of the interaction between neuroendocrine physiology and the immune responses. The rat has been particularly useful in part because, in contrast to other species, most rat models of autoimmune/inflammatory disease are induced rather than spontaneous. This allows for systematic and controlled manipulations of the neuroendocrine system in relation to exposure to the antigen or proinflammatory trigger. The most frequently used immune challenges include lipopolysaccharide-induced septic shock, carrageenan-induced local inflammation and adjuvant or bacterial cell wall-induced arthritis. By analyzing the responses to these challenges in different strains of rats and mice it has been possible to define the relationships between the neuroendocrine and immune systems and to identify some mechanisms through which these connections confer susceptibility and resistance to autoimmune and inflammatory diseases. The present review will discuss data obtained from rodent physiology, indicating that an important component in the susceptibility or resistance to development of these diseases is due to dysfunctional regulation of the immune response by the neuroendocrine hypothalamic-pituitary-adrenal axis. In particular, the importance of neurons of the paraventricular hypothalamic nucleus in determining susceptibility or resistance to autoimmune and inflammatory disease will be discussed. PMID- 11918684 TI - Regulation of inflammatory responses by natural anticoagulants. AB - Proinflammatory cytokines such as tumor necrosis factor-alpha (TNF-alpha) are critically involved in activation of the coagulation system in sepsis, leading to disseminated intravascular coagulation (DIC). Natural anticoagulants such as antithrombin (AT) and activated protein C (APC) regulate the coagulation system by inhibiting thrombin generation. In addition to these anticoagulant effects, both AT and APC have been shown to attenuate inflammatory responses induced by various noxious stimuli in rats such as lipopolysaccharide (LPS) challenge. AT promotes the endothelial release of prostacyclin, a potent anti-inflammatory prostaglandin that inhibits the monocytic production of TNF-alpha, by interacting with cell-surface heparin-like substances. APC directly inhibits the production of TNF-alpha by inhibiting the activation of both nudear factor kappaB (NFkappaB) and activator protein-1 in monocytes stimulated with LPS. Thrombomodulin, an endothelial membranous integral protein that binds thrombin, exerts anti inflammatory effects by generating APC. Furthermore, tissue factor pathway inhibitor, a natural anticoagulant for the extrinsic pathway of the coagulation system, also attenuates LPS-induced inflammatory responses in rats by inhibiting TNF-alpha production by monocytes. These findings strongly suggest that natural anticoagulants could regulate inflammatory responses as well as the coagulation system in rats by inhibiting the monocytic production of TNF-alpha. Such anti inflammatory properties of natural anticoagulants are potentially important for their replacement in patients with sepsis who frequently develop DIC and organ failure as inflammatory responses. PMID- 11918685 TI - Activation of the immune system by bacterial CpG-DNA. AB - The past decade has seen a remarkable process of refocusing in immunology. Cells of the innate immune system, especially macrophages and dendritic cells, have been at the centre of this process. These cells had been regarded by some scientists as non-specific, sometimes perhaps even confined to the menial job of serving T cells by scavenging antigen and presenting it to the sophisticated adaptive immune system. Only over the last few years has it become unequivocally clear that cells of the innate immunity hold, by variation of context and mode of antigen presentation, the power of shaping an adaptive immune response. The innate immune response, in turn, is to a significant degree the result of stimulation by so-called pathogen-associated molecular patterns (PAMPs). One compound with high stimulatory potential for the innate immune system is bacterial DNA. Here we will review recent evidence that bacterial DNA should be ranked with other PAMPs such as lipopolysaccharide (LPS) and lipoteichoic acid. We will further review our present knowledge of DNA recognition and DNA-dependent signal transduction in cells of the immune system. PMID- 11918687 TI - Fas ligand and the fate of antitumour cytotoxic T lymphocytes. PMID- 11918686 TI - Diversity in MHC class II antigen presentation. AB - Processing exogenous and endogenous proteins for presentation by major histocompatibility complex (MHC) molecules to T cells is the defining function of antigen-presenting cells (APC) as major regulatory cells in the acquired immune response. MHC class II-restricted antigen presentation to CD4 T cells is achieved by an essentially common pathway that is subject to variation with regard to the location and extent of degradation of protein antigens and the site of peptide binding to MHC class II molecules. These subtle variations reveal a surprising flexibility in the ways a diverse peptide repertoire is displayed on the APC surface. This diversity may have profound consequences for the induction of immunity to infection and tumours, as well as autoimmunity and tolerance. PMID- 11918688 TI - Immune privilege and FasL: two ways to inactivate effector cytotoxic T lymphocytes by FasL-expressing cells. AB - The theory that Fas ligand (FasL)-expressing tumours are immune-privileged and can directly counterattack Fas-expressing effector T lymphocytes has recently been questioned and several alternative mechanisms have been proposed. To address this controversial issue, we analysed the impact of FasL-expressing tumours on in vivo-primed cytotoxic T lymphocytes (CTLs) and the mechanisms involved. CTLs were obtained from the peritoneal cavity (PEL) after in vivo priming with syngeneic or allogeneic murine tumour cells. We have found that PEL populations undergo Fas based apoptotic cell death when co-cultured with FasL-expressing tumour cells and that PEL destruction of cognate targets in a 51Cr-release assay was markedly inhibited by the pre-exposure to either cognate or non-cognate tumour cells expressing FasL. Furthermore, cytocidal function of PEL was markedly inhibited by preincubation with FasL-negative tumour cells, if and only if they were the cognate targets of the CTL; this CTL inhibition involved FasL-Fas interactions. The killing function of 'bystander' PELs, reactive to a third-party target cell, was inhibited by co-cultivation with PELs mixed with their cognate target. This activation-induced CTL fratricide was not influenced by the expression of FasL on the cognate target cells. These studies demonstrate the existence of two distinct pathways whereby FasL-expressing cells inhibit in vivo-primed FasL- and Fas expressing CTLs: first, by FasL-based direct tumour counterattack, and second, by FasL-mediated activation-induced cell death of the CTLs, which is consistent with the concept that FasL expression in vivo could play a role in inducing immune privilege. PMID- 11918689 TI - Differentiation of human alloreactive CD8(+) T cells in vitro. AB - Expansion and differentiation of alloantigen-reactive CD8(+) T cells in mixed lymphocyte cultures was followed by measurement of the loss of carboxyfluorescein diacetate succinimidyl ester (CFSE) fluorescence of responder cells. Proliferation of CD8(+) T cells became detectable on day 4 of culture and, 2 days later, > 60% of the CD8(+) T cells in culture were dividing alloreactive lymphocytes. In parallel with expansion, CD8(+) T-cell differentiation was initiated, as evidenced by an increase in the number of CD45RA(-) and CD27(-) T cells and acquisition of the ability to produce interferon-gamma after restimulation with the specific alloantigen. Finally, although short-term stimulation and measurement of intracellular cytokine production allowed visualization of alloreactive CD8(+) T cells expanded in vitro, this procedure did not detect circulating alloreactive CD8(+) T cells activated in vivo in recipients of allogeneic kidney grafts. PMID- 11918690 TI - Immunoglobulin A-deficient mice exhibit altered T helper 1-type immune responses but retain mucosal immunity to influenza virus. AB - We have previously demonstrated that immunoglobulin A (IgA)(-/-) knockout (KO) mice exhibit levels of susceptibility to influenza virus infection that are similar to those of their normal IgA(+/+) littermates. To understand the mechanism of this apparent mucosal immunity without IgA, immunoglobulin isotype and T helper 1 (Th1)-type [interferon-gamma (IFN-gamma)] and Th2-type [interleukin (IL)-4, IL-5)] cytokine responses to influenza vaccine were evaluated. Intranasal immunization with influenza virus subunit vaccine plus cholera toxin/cholera toxin B subunit (CT/CTB) induced significant influenza virus-specific immunoglobulin G (IgG) antibody in the serum and nasal passages of both IgA(-/-) and IgA(+/+) mice, while IgA antibodies were induced only in IgA(+/+) mice. IgA KO mice exhibited an IgG1 subclass haemagglutinin (HA) specific response but no detectable IgG2a and IgG2b responses. In contrast, IgA(+/+) mice exhibited significant IgG1 as well as IgG2a responses. This indicates a predominant Th2-type response in IgA KO mice compared to normal mice. Following stimulation with influenza virus in vitro, splenic lymphocytes from immunized IgA(-/-) mice produced significantly lower levels of IFN-gamma than IgA(+/+) mice (P < 0.001), but elaborated similar levels of IL-4 and IL-5. This was true at both protein and mRNA levels. Immunized mice were challenged intranasally with a small inoculum of influenza virus to allow deposition of virus in the nasal mucosal passages. Compared to non-immunized mice, immunized IgA(-/-) and IgA(+/+) mice exhibited significant, but similar levels of reduction in virus titres in the nose and lung. These results demonstrate that in addition to IgA deficiency, IgA gene deletion also resulted in down-regulated Th1-type immune responses and confirm our previous data that IgA antibody is not indispensable for the prevention of influenza virus infection. PMID- 11918691 TI - Identification of broadly recognized, T helper 1 lymphocyte epitopes in an equine lentivirus. AB - Equine infectious anaemia virus (EIAV) is a horse lentivirus causing lifelong, persistent infection. During acute infection, CD8(+) cytotoxic T lymphocytes (CTL) are probably involved in terminating plasma viraemia. However, only a few EIAV CTL epitopes, restricted to fewer horse major histocompatibility complex (MHC) class I alleles, are known. As interferon-gamma (IFN-gamma)-secreting CD4(+), T helper 1 (Th1) lymphocytes promote CTL activity and help maintain memory CTL, identifying broadly recognized EIAV Th1 epitopes would contribute significantly to vaccine strategies seeking to promote strong CTL responses among horses with varying class I haplotypes. To this end, peripheral blood mononuclear cells (PBMC) from 10 MHC disparate, EIAV-infected horses were tested in T lymphocyte proliferation assays for recognition of peptides from the Gag p26 capsid region and a portion of Pol. Both regions are highly conserved among EIAV isolates, and this Pol region is 51-63% homologous to other lentiviral Pol proteins. Seven of 10 horses recognized peptide Gag 221-245, and peptides Gag 242 261 and Pol 323-344 were recognized by five and four horses, respectively. Furthermore, the Gag peptides were recognized by two additional horses after resolving their initial plasma viraemia, indicating that these two peptides can be immunodominant early in infection. Gag peptide-responsive PBMC produced only IFN-gamma, indicating a Th1 response, while Pol 323-344-responsive PBMC produced IFN-gamma both with and without interleukin-4. PBMC from uninfected horses failed to either proliferate or secrete cytokines in response to peptide stimulation. Finally, CD4(+) T lymphocytes were required for proliferation responses, as shown by assays using CD4- versus CD8-depleted PBMC. PMID- 11918692 TI - Conditional ablation of T-cell development by a novel viral ion channel transgene. AB - A novel conditional-lethal transgene system is defined in which a mutated influenza A virus ion-channel protein, which is permeable to monovalent cations, is lethal to cells on heterotypic expression and whose activity can be blocked by an antiviral drug (amantadine), is used to reversibly disrupt T-cell development. In vivo expression of the M2 ion channel, as a transgene under control of the T cell specific p56(Lck) proximal promoter, resulted in total ablation of T-cell development with the accumulation of three distinct populations of early progenitor cells (CD44(+) CD25(-); CD44(+) CD25(+); CD44(+) CD25(hi)) in the thymic rudiment. In vitro development of transgenic fetal thymic progenitors to single-positive T cells could be rescued by antiviral drug treatment. Moreover, there was a radical reduction in B-cell lymphopoiesis, evident at the pre-B-cell stage, with a twofold increase of lymphoid cells 'in cycle' in transgenic bone marrow, indicative of major changes in haematopoietic homeostasis. This system may provide a generic protocol for conditional, lineage-specific cell ablation with available tissue-specific promoters for any eukaryotic developmental system, and provide a window on early T-cell development. PMID- 11918693 TI - Neonatal mycobacterial specific cytotoxic T-lymphocyte and cytokine profiles in response to distinct BCG vaccination strategies. AB - This study evaluated whether different bacillus Calmette-Guerin (BCG) strains, routes of administration, vaccination age and percutaneous tools influenced immune responses to BCG vaccination in infants. Proliferative responses, cytokine production and cell-mediated cytotoxicity obtained in post-vaccinated children were compared to baseline cord bloods and unvaccinated 10-week-old infants. BCG vaccination generally induced strong lymphoproliferative and T helper type 1 (Th1)-type cytokine responses. There was a trend for greater responsiveness following the intradermal route of vaccination, with Japanese-172 strain and with delaying vaccination until 10 weeks. Cord mononuclear cells differentially stimulated the Th2-type cytokines interleukin-5 (IL-5) and IL-10 selectively in response to BCG, as compared to H37Rv or purified protein derivative stimulation. We document for the first time the generation of mycobacterium-specific cytotoxic T lymphocytes in neonates, following BCG vaccination. Cytotoxic activity correlated with the ratio of interferon-gamma to IL-5, aside from a single instance where use of the Biovac tool resulted in a striking dissociation selectively against H37Rv targets. These data have implications for correlates of protective immunity in design of vaccine studies. PMID- 11918695 TI - Mycobacterial infection inhibits established allergic inflammatory responses via alteration of cytokine production and vascular cell adhesion molecule-1 expression. AB - Our previous studies, as well as those of others, have demonstrated that local or systemic Mycobacterium bovis bacille Calmette-Guerin (BCG) infection can inhibit de novo allergen-induced asthma-like reactions, but the effect of this infection on established allergic responses is unknown. The aim of this study was therefore to examine the effect of mycobacterial infection on established allergy in a murine model of asthma-like reaction. Mice were sensitized with ovalbumin (OVA) in alum followed by infection with BCG and subsequent intranasal challenge with the same allergen. In some experiments, mice were sensitized with OVA followed by intranasal challenge with OVA and then given BCG infection with subsequent rechallenge with OVA. Mice without BCG infection but treated with OVA in the same manner, were used as a control. The mice were examined for immunoglobulin E (IgE) response and eosinophilic inflammation, mucus production, cytokine/chemokine patterns and adhesion molecule expression in the lung. The results showed that postallergen BCG infection suppressed the established airway eosinophilia and mucus overproduction, but not IgE responses. The inhibition of asthma-like reactions by BCG infection was correlated with a shift of allergen-driven cytokine production pattern and, more interestingly, with a dramatic decrease of vascular cell adhesion molecule-1 (VCAM-1) expression in the lung. These findings suggest that intracellular bacterial infection can inhibit established allergic responses via alteration of local cytokine production and the expression of adhesion molecules. PMID- 11918694 TI - Distribution of IFN-gamma, IL-4 and TNF-alpha protein and CD8 T cells producing IL-12p40 mRNA in human lung tuberculous granulomas. AB - In order to examine the immune response at the site of pathology in tuberculosis, we analysed cytokines present in lung granulomas, their associations with each other and with caseous necrosis as well as the phenotype of the cellular infiltrate. Paraffin-embedded tissue from the lungs of seven patients with pulmonary tuberculosis was analysed by immunohistochemistry and in situ hybridization to detect interferon-gamma (IFN-gamma), tumour necrosis factor alpha (TNF-alpha) and interleukin-4 (IL-4) proteins and IL-12p40 mRNA. All seven patients had granulomas staining positive for IFN-gamma, TNF-alpha and IL-12p40, but only four stained positive for IL-4. Cells with the morphology of lymphocytes, macrophages and giant cells expressed TNF-alpha, IFN-gamma and IL-4 protein. Furthermore, CD68-positive myeloid cells expressed IL-12p40 mRNA, as expected, but a subset of CD3-positive lymphocytes also expressed this mRNA. These lymphocytes producing IL-12p40 also stained positive for CD8 but not CD4. A total of 141 granulomas were scored for the presence or absence of cytokine or necrosis and two major associations were identified. The first association was between IFN-gamma and IL-12, with 76% of granulomas staining positive for both cytokines. Unexpectedly, those granulomas positive for IL-4 were always positive for IFN-gamma. The second association was between TNF-alpha and caseous necrosis, where all necrotic granulomas were TNF-alpha positive. This association was modulated by IL-4. Therefore, heterogeneity of cellular infiltrate and cytokine expression is observed between adjacent granulomas in the same patient. PMID- 11918696 TI - Lipopolysaccharide triggers invasive streptococcal disease in mice through a tumour necrosis factor-alpha-dependent mechanism. AB - Streptococcus pyogenes sometimes induces invasive streptococcal infection, including streptococcal toxic shock syndrome (STSS). Muscular necrosis is one of the peculiar symptoms of invasive streptococcal infection and STSS. We inoculated S. pyogenes into the muscles of mice. To do so, 5 x 10(8) bacteria in 0.2 ml phosphate-buffered saline were injected into the right hind thigh. None of the mice injected with the bacteria showed muscular necrosis and none died. Tumour necrosis factor-alpha (TNF-alpha) and infiltration of leucocytes were detected in the muscles of infected sites, although the condition of the infected mice did not deteriorate after anti-TNF-alpha monoclonal antibody treatment. The infected mice treated intraperitoneally with Escherichia coli lipopolysaccharide (LPS) showed augmentation of bacterial growth, muscular necrosis and death. TNF-alpha was detected in the sera of the infected mice treated with LPS, but not in the muscles of the infected sites. Infiltration of leucocytes into the infected muscle was not observed in the infected mice treated with LPS. Anti-TNF-alpha monoclonal antibody treatment decreased mortality in the infected mice treated with LPS. Moreover, the infected mice treated with recombinant TNF-alpha showed augmentation of muscular necrosis and death. These results suggest that systemic production of TNF-alpha induced by stimulation with LPS inhibits infiltration of leucocytes into the infected site and exacerbates muscular infection, and that TNF-alpha produced in streptococcal infection is not a defence factor for the host. Invasive streptococcal infection and STSS appear to be induced by both S. pyogenes and the host's immune system. PMID- 11918700 TI - Vitamin C regulates keratinocyte viability, epidermal barrier, and basement membrane in vitro, and reduces wound contraction after grafting of cultured skin substitutes. AB - Cultured skin substitutes have become useful as adjunctive treatments for excised, full-thickness burns, but no skin substitutes have the anatomy and physiology of native skin. Hypothetically, deficiencies of structure and function may result, in part, from nutritional deficiencies in culture media. To address this hypothesis, vitamin C was titrated at 0.0, 0.01, 0.1, and 1.0 mM in a cultured skin substitute model on filter inserts. Cultured skin substitute inserts were evaluated at 2 and 5 wk for viability by incorporation of 5-bromo-2' deoxyuridine (BrdU) and by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT) conversion. Subsequently, cultured skin substitute grafts consisting of cultured human keratinocytes and fibroblasts attached to collagen glycosaminoglycan substrates were incubated for 5 wk in media containing 0.0 mM or 0.1 mM vitamin C, and then grafted to athymic mice. Cultured skin substitutes (n = 3 per group) were evaluated in vitro at 2 wk of incubation for collagen IV, collagen VII, and laminin 5, and through 5 wk for epidermal barrier by surface electrical capacitance. Cultured skin substitutes were grafted to full-thickness wounds in athymic mice (n = 8 per group), evaluated for surface electrical capacitance through 6 wk, and scored for percentage original wound area through 8 wk and for HLA-ABC-positive wounds at 8 wk after grafting. The data show that incubation of cultured skin substitutes in medium containing vitamin C results in greater viability (higher BrdU and MTT), more complete basement membrane development at 2 wk, and better epidermal barrier (lower surface electrical capacitance) at 5 wk in vitro. After grafting, cultured skin substitutes with vitamin C developed functional epidermal barrier earlier, had less wound contraction, and had more HLA-positive wounds at 8 wk than without vitamin C. These results suggest that incubation of cultured skin substitutes in medium containing vitamin C extends cellular viability, promotes formation of epidermal barrier in vitro, and promotes engraftment. Improved anatomy and physiology of cultured skin substitutes that result from nutritional factors in culture media may be expected to improve efficacy in treatment of full-thickness skin wounds. PMID- 11918698 TI - A feline CD2 homologue interacts with human red blood cells. AB - A cDNA encoding a feline homologue of CD2 (fCD2) was identified. Several amino acids (aa) important for ligand interaction, molecular folding or signal transduction, found in other mammalian CD2, were found to be highly conserved in the predicted fCD2 aa sequence. fCD2-expressing cells were able to form rosettes with human red blood cells (probably via human CD58), and the rosette formation was inhibited by an anti-fCD2 monoclonal antibody. These results are indicative of the similarity of feline and human CD2 structures. fCD2 was found to be expressed in feline peripheral blood T lymphocytes, monocytes and cultured lymphoid cells. PMID- 11918697 TI - Synergistic interactions of the two classes of ligand, sialyl-Lewis(a/x) fuco oligosaccharides and short sulpho-motifs, with the P- and L-selectins: implications for therapeutic inhibitor designs. AB - The E-, L- and P-selectins are carbohydrate-recognizing cell-adhesion molecules mediating selective leucocyte recruitment in inflammation. The 3'-sialyl- and 3' sulpho-oligosaccharides of Lewis(x) (Le(x)) and Lewis(a) (Le(a)) series are bound by them, but for high-avidity binding of P- and L-selectins to the glycoprotein counter-receptor known as P-selectin glycoprotein ligand, PSGL-1, there is a requirement for sulpho-tyrosines neighbouring a sialyl-Le(x) glycan. The two selectins can also bind 3-O- or 6-O-sulphated galacto-lipids (sulphatides). Here we compare some features of the interactions of P- and L-selectins with a novel lipid-linked sulpho-tyrosine probe, and with the sulphatides and neoglycolipids of sialyl- and sulpho-Le(x)/Le(a) fuco-oligosaccharides. The sulpho-tyrosine probe is bound by both selectins. There are close similarities in the interactions of the two selectins with sulpho-tyrosine and the sulphatides; the binding is relatively resistant to chelation of calcium ions, in contrast to the absolute requirement of calcium ions with the long fuco-oligosaccharides, including 6-sulpho-sialyl-Le(x). With both selectins, there is striking synergy in binding signals elicited by the two ligand types when presented as equimolar mixtures on a matrix. Thus, there are two operationally distinct binding sites on both L- and P-selectin; and the binding sites for sulphate groups in the two ligand types are probably distinct. When sulpho-tyrosine and sialyl-Le(x) are presented on liposomes, a potent inhibitory activity is generated toward the binding of P-selectin to HL60 cells, with 50% inhibitory concentration (IC(50)) values in the nanomolar range. These features of the lipid-linked ligand analogues, and the simple approach for their display on liposomes, may have applications in designs and screening of selectin inhibitors as anti-inflammatory compounds. PMID- 11918701 TI - Membrane type 1 matrix metalloproteinase regulates cellular invasiveness and survival in cutaneous epidermal cells. AB - Membrane type 1 matrix metalloproteinase is a member of the membrane-anchored matrix metalloproteinase family and is involved in tissue remodeling events ranging from tumor invasion and angiogenesis to growth and development. We sought to clarify the role of membrane type 1 matrix metalloproteinase in cutaneous epidermal cells using anti-sense cDNA expression in human keratinocytes. Modulation of membrane type 1 matrix metalloproteinase transcript and protein levels was achieved via retroviral expression of a 5' 1.4 kb anti-sense membrane type 1 matrix metalloproteinase construct and a 3.4 kb full-length sense membrane type 1 matrix metalloproteinase construct in primary and immortalized keratinocytes and SCC-25 cells. Maximal reductions were observed 48-72 h after transduction with 1.4 kb anti-sense membrane type 1 matrix metalloproteinase construct that correlated with significant decreased pro-matrix metalloproteinase 2 activation. Functionally, we found decreased cell migration, reduced cellular proliferation, and increased apoptotic nuclear fragmentation after 1.4 kb anti sense membrane type 1 matrix metalloproteinase construct expression. Our findings suggest a role for membrane type 1 matrix metalloproteinase in human cutaneous epidermal cell invasion and survival mechanisms in vivo. PMID- 11918702 TI - Cholesterol-rich plasma membrane domains (lipid rafts) in keratinocytes: importance in the baseline and UVA-induced generation of reactive oxygen species. AB - The biologic effects of ultraviolet radiation such as DNA damage, mutagenesis, cellular aging, and carcinogenesis are in part mediated by reactive oxygen species. In unirradiated cells the major known sources of reactive oxygen species are the mitochondrial respiratory chain and the membrane oxidases functionally coupled to several membrane growth factor receptors. There is evidence that mitochondria also play a role in oxidative stress after ultraviolet irradiation; however, it is unknown whether the biochemical processes at the level of the plasma membrane contribute to the regulation of reactive oxygen species synthesis. In order to elucidate this issue we examined here the importance of the microdomain plasma membrane organization in the regulation of oxidative stress in unirradiated and ultraviolet A (340-400 nm) irradiated HaCaT keratinocytes. Labeling of confluent HaCaT cultures with fluorescently tagged cholera toxin B subunit (FITC-CTx) revealed the presence of GM1 ganglioside and cholesterol-rich microdomains (lipid rafts) that formed junction-like structures in the membranes of adjacent cells and patchy microdomains elsewhere. There was a marked heterogeneity in the level of FITC-CTx labeling: there were groups of cells demonstrating prominent labeling (FITC-CTx(high)) whereas other cells were only weakly labeled (FITC-CTx(low)). When reactive oxygen species synthesis was measured with the fluorescent probe carboxy-2',7'-dichlorodihydrofluorescein diacetate, we found that (i) the baseline and ultraviolet-A-induced reactive oxygen species synthesis correlated with the magnitude of FITC-CTx labeling and was highest in the FITC-CTx(high) cells; (ii) reactive oxygen species synthesis was diminished in cells in which the integrity of membrane domains was disrupted by cholesterol sequestration with methyl-beta-cyclodextrin and filipin, or after treatment with GM1 ganglioside; (iii) reactive oxygen species synthesis in cholesterol-depleted cells was fully restored after cholesterol repletion. We conclude that the plasma membrane takes part in the regulation of oxidative stress in keratinocytes and disruption of its microdomain structure reduces reactive oxygen species synthesis both at the baseline and after ultraviolet A irradiation. We hypothesize that lipid-raft-associated protein(s) may be involved in the generation of reactive oxygen species and that pharmacologic modulation of membrane structure may provide a novel therapeutic approach relevant for photoprotection and cutaneous carcinogenesis. PMID- 11918703 TI - The ENaC channel is required for normal epidermal differentiation. AB - Ionic fluxes are important for critical aspects of keratinocyte differentiation, including synthesis of differentiation-specific proteins, enzymatic catalysis of protein cross-linking, post-transcriptional processing of profilaggrin, and lipid secretion. The epithelial sodium channel is expressed in epidermis and the expression of its alpha and beta subunits is enhanced as keratinocytes differentiate. In order to ascertain the role of the epithelial sodium channel in epidermal differentiation, we examined skin of mice in which the epithelial sodium channel alpha subunit had been deleted. Newborn -/- mice, in which the alpha subunit had been completely inactivated, demonstrated epithelial hyperplasia, abnormal nuclei, premature secretion of lipids, and abnormal keratohyaline granules. In addition, immunohistochemistry demonstrated that expression of the differentiation markers K1, K6, and involucrin were abnormal. These data suggest that the epithelial sodium channel modulates ionic signaling for specific aspects of epidermal differentiation, such as synthesis or processing of differentiation- specific proteins, and lipid secretion. PMID- 11918704 TI - Differential expression of inhibitory or activating CD94/NKG2 subtypes on MART-1 reactive T cells in vitiligo versus melanoma: a case report. AB - Selection and activation of T cells is tightly regulated by both antigen-specific receptors and co-receptors to ensure that responses to self antigens are largely avoided. By T cell receptor clonotypic mapping and staining with tetrameric HLA peptide complexes, we demonstrate the presence of melanocyte differentiation antigen MART-1 specific T cells in the areas of destruction of both neoplastic and normal melanocytic cells in a case of a primary melanoma and its associated hypopigmentation. These self reactive T cells expressed CD94/NKG2 major histocompatibility complex class I specific C-type lectin-like receptors. This family of receptors includes both activating and inhibitory isoforms. Thus, we performed a detailed analysis that revealed the exclusive presence of inhibitory NKG2-A/B receptors in the vitiligo-like leukoderma, whereas both the inhibitory receptors and the activating NKG2-C/E isoforms were present within the tumor. Our data suggest the differential expression of killer inhibitory receptors as a possible mechanism to regulate T cell responses to self antigens. PMID- 11918705 TI - Expression of stress-induced MHC class I related chain molecules on human melanoma. AB - Cellular immune responses to melanoma are tightly regulated and include specific T cell responses to self antigens such as Mart-1 and gp100. Thus, additional signals apart from those mediated by the T cell receptor are needed to ensure T cell activation. Recently, the stress inducible major histocompatibility complex molecules, MHC class I related chain, were identified as an activator of both natural killer and T cells via interaction with their receptor NKG2D. Herein, we report the expression of MIC in 31 of 40 primary cutaneous melanomas and in 13 of 20 metastatic lesions. Moreover, lymphocytes infiltrating the tumor were found to express NKG2D. Detailed analysis identified both CD3+ T cells as well as CD56+ natural killer cells contributing to this NKG2D+ tumor infiltrating lymphocyte population present. PMID- 11918706 TI - Phase behavior of stratum corneum lipid mixtures based on human ceramides: the role of natural and synthetic ceramide 1. AB - In a recent study the lipid phase behavior of mixtures of human ceramides, cholesterol, and free fatty acids has been examined. We observed in cholesterol: human ceramide mixtures a prominent formation of the 12.8 nm lamellar phase (referred to as the long periodicity phase). Addition of free fatty acids promoted the formation of a 5.6 nm lamellar phase (referred to as the short periodicity phase) and increased the subpopulation of lipids forming a fluid phase. In this study we focused on the role of human ceramide 1, as the presence of this ceramide appeared to be crucial for proper lipid phase behavior in mixtures prepared with ceramide isolated from pig stratum corneum. In order to do this, mixtures of cholesterol and free fatty acids were prepared with human ceramides, in which natural human ceramide 1 was replaced by either synthetic CER1-linoleate (CER1-lin), or CER1-oleate (CER1-ol), or CER1-stearate (CER1-ste). After substitution of natural human ceramide 1 by synthetic ceramide 1 the following observations were made. (i) In the presence of synthetic CER1-ste no long periodicity phase and no liquid phase could be detected. (ii) In the presence of HCER1-ol a liquid phase was more prominently formed than in the presence of HCER1-lin. (iii) In cholesterol:human ceramide mixtures in the presence of CER1-lin the long periodicity phase was more prominently present than in the presence of CER1-ol. (iv) In the presence of CER1-ste neither a long periodicity phase nor a liquid lateral packing could be detected. The results of these studies further indicate that for the formation of the long periodicity phase a certain (optimal) fraction of lipids has to form a liquid phase. When the fraction forming this liquid phase is either too low or too high, the formation of the short periodicity phase is increased at the expense of the formation of the long periodicity phase. Based on the results of this and previous studies we offer an explanation for the deviation in lipid organization in diseased and in dry skin compared to normal skin. PMID- 11918707 TI - Photoaging is associated with protein oxidation in human skin in vivo. AB - There is increasing evidence for the generation of reactive oxygen species in skin upon ultraviolet exposure, but little is known about their pathophysiologic relevance in human skin in vivo. We hypothesized that chronic and acute photodamage is mediated by depleted antioxidant enzyme expression and increased oxidative protein modifications. Biopsies from patients with histologically confirmed solar elastosis, from non-ultraviolet-exposed sites of age-matched controls, and from young subjects were analyzed. To evaluate the influence of acute ultraviolet exposures, buttock skin of 12 healthy subjects was irradiated repetitively on 10 d with a solar simulator and compared intraindividually to non ultraviolet-treated contralateral sites. The antioxidant enzymes catalase, copper zinc superoxide dismutase, and manganese superoxide dismutase were investigated by immunohistochemistry. Protein carbonyls were analyzed by immunohistochemical and immunoblotting techniques in human skin and in cell models. Whereas overall expression of antioxidant enzymes was very high in the epidermis, low baseline levels were found in the dermis. In photoaged skin, a significant depletion of antioxidant enzyme expression was observed within the stratum corneum and in the epidermis. Importantly, an accumulation of oxidatively modified proteins was found specifically within the upper dermis of photoaged skin. Upon acute ultraviolet exposure of healthy subjects, depleted catalase expression and increased protein oxidation were detected. Exposures of keratinocytes and fibroblasts to ultraviolet B, ultraviolet A, and H2O2 led to dose-dependent protein oxidation and thus confirmed in vivo results. In conclusion, the correlation between photodamage and protein oxidation was demonstrated for the first time, which hence may be a relevant pathophysiologic factor in photoaging. PMID- 11918708 TI - Partial revertant mosaicism of keratin 14 in a patient with recessive epidermolysis bullosa simplex. AB - A patient with recessive epidermolysis bullosa simplex due to a previously described homozygous KRT14 1842-2A-->C splice-site mutation was re-examined, because we unexpectedly found signs of revertant mosaicism. The germline mutation resulted in different aberrant transcripts containing premature termination codons, all leading to truncated keratin 14 proteins. Basal keratinocytes in skin and in culture completely lacked keratin 14 and intermediate filaments. From this keratin 14-/- patient we started cultures from a new skin biopsy and here, we serendipitously found keratinocytes that spontaneously expressed keratin 14. This biopsy had been taken from an area of skin that was clinically affected, because blisters could simply be evoked by gentle rubbing. Immunofluorescence and electron microscopy of additional biopsies from this skin area revealed a mosaic expression of keratin 14 and reappearance of intermediate filaments in basal keratinocytes. Immunoblotting showed a revertant keratin 14 polypeptide with seemingly normal molecular weight. DNA analysis of exon 2 and its flanking intron borders showed no additional mutations in the genomic KRT14 sequence. Analysis of mRNA isolated from mosaic skin keratinocytes revealed an additional in-frame transcript (1844T-->G, 1845Delta6) that codes for an abnormal keratin 14 polypeptide with a two residue deletion and one amino acid change. The re expression of a revertant, albeit abnormal, keratin 14 polypeptide, so-called partial revertant mosaicism, accounts for the antibody staining pattern and for the reappearance of intermediate filaments, which however, are semifunctional and not able to revert the clinical phenotype. The combination of a keratin 14 positive and a keratin 14-negative cell population in epidermis as well as in cultured keratinocytes suggests that the cellular reversion might be caused by an endogenous factor. We hypothesize that a second somatic modulating factor in the genome that affects the processing of the mutant KRT14 pre-mRNA may underlie this phenomenon. PMID- 11918709 TI - Targeted expression of human vitamin d receptor in the skin promotes the initiation of the postnatal hair follicle cycle and rescues the alopecia in vitamin D receptor null mice. AB - Alopecia is a predominant feature of vitamin D receptor inactivation in mice and humans. To determine the role of vitamin D receptor in the regulation of hair growth directly, we used the human keratin 14 promoter to target human vitamin D receptor expression to the skin of transgenic mice, and generated vitamin D receptor null mice that express the human vitamin D receptor transgene. Parallel studies were carried out in littermates of wild-type, vitamin D receptor null, transgenic, and human vitamin D receptor-expressing null mice in two transgenic lines. The transgenic mice were grossly normal. The vitamin D receptor null and vitamin D receptor null/human vitamin D receptor mice were growth retarded and developed hypocalcemia, secondary hyperparathyroidism, and rickets. In contrast to the vitamin D receptor null mice that developed alopecia, however, the vitamin D receptor null/human vitamin D receptor mice displayed a normal hair coat, and their hair shaft and skin histology were indistinguishable from those of the wild type mice. Immunohistochemical analyses revealed that the human vitamin D receptor was highly expressed in the basal layer of the epidermis and outer root sheath of the hair follicle. During follicular morphogenesis, no major histologic differences were seen in the skin of wild-type, vitamin D receptor null, transgenic, and vitamin D receptor null/human vitamin D receptor littermates. When anagen was induced by hair depilation at day 20 after birth, the vitamin D receptor null mice failed to initiate the hair cycle, whereas the vitamin D receptor null/human vitamin D receptor mice displayed the same pattern of anagen follicle formation as the wild-type mice. Interestingly, the transgenic mice initiated the follicular cycle earlier than the wild-type and vitamin D receptor null/human vitamin D receptor mice in a gene concentration-dependent manner. Taken together, these data provide direct evidence that vitamin D receptor is required for the initiation of the postnatal hair follicular cycle in mice. PMID- 11918710 TI - PTEN mutations in eight Spanish families and one Brazilian family with Cowden syndrome. AB - Cowden syndrome is an autosomal dominant genodermatosis, characterized by the presence of multiple hamartomas in the skin, breast, thyroid, gastrointestinal tract, central nervous system, and an increased risk in developing breast and thyroid carcinomas. Over 80 germline mutations of the tumor suppressor gene PTEN, on chromosome 10q23, have been reported in more than 100 unrelated patients and families; however, questions regarding distribution of the mutations in populations from different geographic areas, and phenotypic expression are still unclear. In this study the results are reported of mutation analysis of PTEN in 13 families from Spain and one family of Brazilian origin with Cowden syndrome. PTEN germline mutations were detected in nine of them (64%). Five mutations were located in exon 5, one in exon 6, two in exon 7, and one in exon 8. Four of the mutations were novel. In another case, an identical change had been previously reported as a somatic mutation in an endometrial carcinoma. In one family, the patient presented a de novo mutation, which was not detected in his parents. In five patients, the detection of the PTEN germline mutation confirmed their condition, even in the absence of sufficient criteria to make the clinical diagnosis of Cowden syndrome. PMID- 11918711 TI - Infection with influenza a virus leads to flu antigen-induced cutaneous anaphylaxis in mice. AB - It is well established, that viral infections may trigger urticaria or allergic asthma; however, as viral infections induce T helper 1 polarized responses, which lead to the inhibition of T helper 2 cell development, the opposite would be plausible. We wanted to investigate how viral infections may mediate allergic symptoms in a mouse model; therefore, we infected BALB/C mice with influenza A virus intranasally. Histologic analyses of lung sections and bronchoalveolar lavages were performed. In addition, cells from the mediastinal lymph nodes were restimulated in vitro to analyze which types of cytokines were induced by the flu infection. Furthermore, flu-specific antibody titers were determined and local anaphylaxis was measured after rechallenge with flu antigen. We found that airways inflammation consisted predominately of macrophages and lymphocytes, whereas only a few eosinophils were observed. interferon-gamma but no interleukin 4 and little interleukin-5 could be detected in the culture supernatants from in vitro restimulated T cells from the draining lymph nodes. The antibody response was characterized by high levels of virus-specific IgG2a, IgG2b, and IgG1 and, surprisingly, low levels of virus-specific IgE antibodies. Interestingly, flu infected mice developed active and passive cutaneous anaphylaxis after rechallenge with flu-antigen. As the passive cutaneous anaphylaxis reaction persisted over 48 h and was significantly lower after passive transfer of the serum, which was IgE depleted, local anaphylaxis seemed to be mediated predominately by specific IgE antibodies. Taken together, our results demonstrate that mice infected with flu virus develop virus-specific mast cell degranulation in the skin. Our results may also have implications for the pathogenesis of urticaria or other atopic disorders in humans. PMID- 11918712 TI - Infection of human oral epithelia with Candida species induces cytokine expression correlated to the degree of virulence. AB - A defined and balanced immunomodulatory response is crucial for the protection of mucosal surfaces being in contact with pathogenic microorganisms. This study examined the local host response mechanisms of epithelial cells in experimental Candida albicans, C. tropicalis, and C. glabrata infections by measuring the expression of cytokines at the mRNA and protein level. During the course of infection with active but not with heat-killed C. albicans stimulation of the gene expression levels for interleukin-1alpha, interleukin-1beta, tumor necrosis factor, Exodus-2, P-selectin ligand, granulocyte-monocyte colony-stimulating factor, and interleukin-8 was observed by standard and quantitative reverse transcription-polymerase chain reaction. This cytokine pattern may favor a chemotactic and a T helper 1 response. Initial moderate or weak upregulation of these cytokine genes by reverse transcription-polymerase chain reaction was also observed in epithelial infection with the less virulent species C. tropicalis and C. glabrata. Heat-killed C. albicans failed to induce an epithelial immune response. At the protein level, expression of interleukin-8 protein was strongly enhanced during the course of C. albicans infection, whereas lower levels were seen with C. tropicalis and C. glabrata. The different expression patterns of cytokines were associated with differences in virulence of the Candida strains. This study's data, therefore, show a correlation between the virulence potential of pathogenic fungi, possibly mediated by specific virulence factors (such as proteinases), and the secretion of epithelial cytokines and chemokines, which may initiate in vivo a protective T helper 1 immunologic response and contribute to the recruitment of activated leukocytes and lymphocytes to the site of mucosal infection. PMID- 11918713 TI - Increased expression of endothelial cell adhesion molecules due to mediator release from human foreskin mast cells stimulated by autoantibodies in chronic urticaria sera. AB - Histamine-releasing antibodies that act against the epitope of the alpha chain of Fc(epsilon)RI (anti-Fc(epsilon)RI(alpha) antibody) that may affect pathogenesis in serum of patients with chronic urticaria. We assessed the capability of anti Fc(epsilon)RI(alpha) antibody in sera from patients with chronic urticaria to release histamine and cytokines, and to induce the expression of endothelial cell adhesion molecules. We also assessed the release of inflammatory mediators from cultured foreskin mast cells, and expression of endothelial cell adhesion molecules on human dermal microvascular endothelial cells. Cells were pretreated with mast cell-conditioned media: culture media of mast cells treated with sera from chronic urticaria patients containing anti-Fc(epsilon)RI(alpha) antibody. Histamine release from human foreskin mast cells challenged with sera, increased after both 20 min and 16 h intervals. Leukotriene D4 release also increased at both 20 min and 16 h. Tumor necrosis factor-alpha increased significantly in foreskin mast cell culture challenged with sera of chronic urticaria patients. After the stimulation of human dermal microvascular endothelial cells with the conditioned media, the expression of intercellular cell adhesion molecule-1, vascular cell adhesion molecule-1, and E-selectin increased significantly. Treatment of the conditioned media with anti-tumor necrosis factor-alpha monoclonal antibody partially inhibited the expression of intercellular cell adhesion molecule-1, vascular cell adhesion molecule-1, and E-selectin. The data suggest that sera from patients with chronic urticaria containing anti Fc(epsilon)RI(alpha) antibody release mediators and tumor necrosis factor-alpha by activating human foreskin mast cells. This release can play a pathogenic role in chronic urticaria by activating endothelial cells, in part due to the actions of tumor necrosis factor-alpha from mast cells. PMID- 11918714 TI - Autoantibodies to bullous pemphigoid antigen 180 induce dermal-epidermal separation in cryosections of human skin. AB - Bullous pemphigoid is a subepidermal autoimmune blistering disease associated with autoantibodies to the hemidesmosomal bullous pemphigoid antigens 180 and 230. Most sera from bullous pemphigoid patients recognize epitopes within the N terminal NC16A portion of the bullous pemphigoid 180 ectodomain. Using cryosections of human skin, patients' sera were shown to generate dermal epidermal separation when coincubated with leukocytes and complement from healthy volunteers; however, the specificity of pathogenic autoantibodies in bullous pemphigoid patients has not yet been elucidated. In this study, by the use of a modified version of the cryosection model, we show that sera from all of 13 bullous pemphigoid patients and from two rabbits, immunized against bullous pemphigoid 180 NC16A, induced dermal-epidermal separation. This finding was confirmed with the use of IgG purified from patients' sera, whereas sera and purified IgG from healthy controls were not pathogenic. The induction of subepidermal splits in this experimental model was shown to be dependent on the presence of neutrophils, but not complement. Interestingly, patients' autoantibodies affinity purified against a recombinant form of bullous pemphigoid 180 NC16A retained their blister-inducing capacity, whereas patients' IgG depleted of reactivity to NC16A lost this ability. F(ab')2 fragments of antibodies specific to NC16A, lacking the Fc portion, did not induce splits. In addition, patients' autoantibodies purified against a recombinant fragment of the C-terminus of bullous pemphigoid 180 as well as rabbit antibodies to the intracellular portion of bullous pemphigoid 180 and to bullous pemphigoid 230 did not cause dermal-epidermal separation. Our in vitro results support the idea that autoantibodies to bullous pemphigoid 180 from patients with bullous pemphigoid are of pathogenic relevance. PMID- 11918715 TI - Immunomodulation by interleukin-10 therapy decreases the incidence of relapse and prolongs the relapse-free interval in Psoriasis. AB - The ability of interleukin-10 therapy to reduce the severity of exacerbated psoriasis has been demonstrated recently. Considering the immunobiologic properties of this cytokine we investigated the effects of long-term interleukin 10 application on the immune system and duration of psoriasis remission. We performed a placebo-controlled, double-blind, phase II trial using interleukin-10 in patients with chronic plaque psoriasis in remission. Patients received subcutaneous injections with either interleukin-10 (10 microg per kg body weight; n = 7) or placebo (n = 10) three times per week until relapse or study termination after 4 months. The treatment was well tolerated. In the placebo group almost all patients (90%) showed a relapse during the observation period. In contrast to this, only two of seven patients (28.6%) relapsed in the interleukin-10-treated group. Kaplan-Meier analysis revealed a significantly lower relapse incidence in the interleukin-10 than in the placebo group (p = 0.02). The mean relapse-free interval time was 101.6 +/- 12.6 d in the interleukin-10 group in comparison with 66.4 +/- 10.4 d in the placebo group. Immunologic activity of interleukin-10 application was indicated by an increase in soluble interleukin-2 receptor plasma levels and higher ex vivo interleukin-4 secretion capacities. Remarkably, a significant negative correlation was demonstrated between the interleukin-4 secretion capacity and Psoriasis Area and Severity Index score (r = -0.36, p < 0.01). Our data suggest that interleukin-10 therapy is immunologic effective, decreases the incidence of relapse and prolongs the disease-free interval in psoriasis. Its value should be further determined in larger trials and for the prevention of re-exacerbation of other inflammatory disorders with a similar immunologic profile. PMID- 11918716 TI - Functional expression of AQP3 in human skin epidermis and reconstructed epidermis. AB - The purpose of this study was to examine the presence of aquaporin water channels in human skin and to assess their functional role. On western blots of human epidermis obtained from plastic surgery, a strong signal was obtained with polyclonal anti-aquaporin-3 antibodies. By indirect immunofluorescence on 5 microm cryosections, anti-aquaporin-3 antibodies strongly stained keratinocyte plasma membranes in human epidermis, whereas no staining was observed in the dermis or the stratum corneum or when anti-aquaporin-3 antibodies were preabsorbed with the peptide used for immunization. Similarly, a strong signal with anti-aquaporin-3 antibodies was observed in keratinocyte plasma membranes of reconstructed human epidermis in culture at the air-liquid interface for up to 3 wk. The keratinocyte plasma membrane localization of aquaporin-3 was confirmed at the electron microscope level in prickle cells. In addition an intracellular localization of aquaporin-3 was also detected in epidermis basal cells. Osmotically induced transepidermal water permeability was measured on stripped human skin and on reconstructed epidermis. Water transport across both stripped human skin and 2-3 wk reconstructed epidermis was comparable, inhibited by > 50% by 1 mM HgCl2 and fully inhibited by acid pH. By stopped-flow light scattering, keratinocyte plasma membranes, where aquaporin-3 is localized, exhibited a high, pH-sensitive, water permeability. Although human skin is highly impermeable to water, this is primarily accounted for by the stratum corneum, where a steep water content gradient was demonstrated. In contrast, the water content of viable strata of the epidermis is remarkably constant. Our results suggest that the human epidermis, below the stratum corneum, exhibits a high, aquaporin-3 mediated, water permeability. We propose that the role of aquaporin-3 is to water clamp viable layers of the epidermis in order to improve the hydration of the epidermis below the stratum corneum. PMID- 11918717 TI - MICA gene polymorphism is not associated with an increased risk for skin cancer. AB - The MICA gene encodes for major histocompatibility complex class I chain-related proteins (MIC), which belong to a recently identified new family of nonclassical major histocompatibility complex molecules. The general structure of the MICA molecule resembles that of major histocompatibility complex class I molecules. MIC molecules are considered to be stress-induced antigens that are recognized by cytotoxic T cells and natural killer cells, which play an important role in the surveillance of transformed infected and damaged cells. Associations of major histocompatibility complex class I molecules with skin cancer have been described before. To evaluate the possible association of MICA gene polymorphism with the risk for nonmelanoma skin cancer we evaluated 153 cases with squamous cell carcinoma, 261 cases with basal cell carcinoma, 111 controls with malignant melanoma, and 247 controls without a history of skin cancer. Five distinct MICA alleles A4, A5, A6, A9, and A5.1 were studied. As the MICA 5.1 variant gene contains a four-nucleotide insertion that causes a stop codon in the trans membrane region, the resulting truncated MICA molecule does not reside on the cellular membrane. In the case of individuals who are homozygous for MICA 5.1 this results in cells that are naked for the MICA molecule. We therefore specifically addressed the possible association between MICA 5.1 homozygosity and skin cancer, as these individuals are expected to be at the highest risk for skin cancer if the MICA gene plays a role in skin carcinogenesis. Viral proteins may serve as antigens for recognition of skin cancer by the immune system. Human papillomavirus is the most likely candidate virus to be involved in the carcinogenesis of cutaneous squamous cell carcinoma. Hence, we also assessed the association between MICA polymorphism and squamous cell carcinoma in human papillomavirus-positive and human-papillomavirus-negative individuals as identified by the presence of human papillomavirus DNA in hairs plucked from their eyebrows. Our analyses did not reveal any significant differences regarding the MICA allele frequencies between cases and controls. Also homozygotes and heterozygotes for the MICA 5.1 variant gene were not at an increased risk for skin cancer compared to individuals without this variant gene and infection with human papillomavirus did not materially influence these findings. The same group of cases and controls was large enough to show an association between melanocortin 1 receptor gene polymorphism and skin cancer and to reasonably exclude an association between p53 codon 72 polymorphism and skin cancer. Therefore, we conclude that an association between MICA gene polymorphism and nonmelanoma skin cancer is not likely. PMID- 11918718 TI - Eicosapentaenoic acid, a n-3 polyunsaturated fatty acid differentially modulates TNF-alpha, IL-1alpha, IL-6 and PGE2 expression in UVB-irradiated human keratinocytes. AB - In response to UVB-irradiation keratinocytes release a variety of cytokines and prostaglandins, including tumor necrosis factor alpha (TNF-alpha), interleukin 1alpha (IL-1alpha), interleukin 6 (IL-6), and prostaglandin E2 (PGE2). n-3 Polyunsaturated fatty acids (PUFA), mainly present in fish oil, can modulate cytokine synthesis, as predominantly studied in macrophages. In order to investigate the immune modulating actions of n-3 PUFA on the UVB response in human skin, we investigated the effect of eicosapentaenoic acid (EPA), a n-3 PUFA and a precursor of eicosanoid biosynthesis, on UVB-modulated TNF-alpha, IL 1alpha, IL-6, and PGE2 expression in normal human keratinocytes (NHK). We show that cultured NHK can efficiently take up EPA. Basal TNF-alpha expression is very low in NHK. IL-1alpha on the contrary is significantly present in untreated cultured NHK. Upon UVB-irradiation (32 mJ per cm2) TNF-alpha mRNA expression and secretion is induced and IL-1alpha mRNA expression is reduced, although IL-1alpha secretion is induced. EPA treatment results in higher TNF-alpha and IL-1alpha expression, both in nonirradiated and UVB-irradiated keratinocytes. Moreover EPA and UVB appear to act synergistically to superinduce TNF-alpha expression. EPA treatment results also in lipid peroxidation and in decreased PGE2 and IL-6 secretion after UVB-irradiation. In contrast to EPA, oleic acid (monounsaturated fatty acid) and linoleic acid (n-6 PUFA) treatment did not result in higher TNF alpha or IL-1alpha levels in nonirradiated or UVB-irradiated NHK, indicating that the observed effects are specific for EPA. In conclusion, these results show that EPA can differentially modulate UVB-induced cytokine and prostaglandin synthesis in NHK. PMID- 11918719 TI - Impaired neurogenic control of skin perfusion in erythromelalgia. AB - Erythromelalgia is a clinical diagnosis characterized by erythema, increased temperature and burning pain in acral skin. The pain is relieved by cooling and aggravated by warming. The symptoms have been hypothesized to be caused by skin hypoxia due to increased arteriovenous shunting. We examined skin microvascular perfusion in response to vasoconstrictory and vasodilatory stimuli, to characterize local and central neurogenic reflexes as well as vascular smooth muscle and vascular endothelial function, using laser Doppler perfusion measurements in 14 patients with primary erythromelalgia and healthy control persons. Skin perfusion preceding provocative stimuli was significantly reduced in patients with erythromelalgia (p < 0.01). The laser Doppler flowmetry signal after sympathetic stimulation of reflexes mediated through the central nervous system, was significantly diminished in patients with erythromelalgia as compared with healthy controls (Valsalva's maneuver p < 0.01; contralateral cooling test p < 0.05). Local neurogenic vasoconstrictor (venous cuff occlusion and dependency of the extremity) and vasodilator reflexes (local heating of the skin), as well as tests for vascular smooth muscle and vascular endothelial function (postocclusive hyperemic response) were maintained. These results indicate that postganglionic sympathetic dysfunction and denervation hypersensitivity may play a pathogenetic role in primary erythromelalgia, whereas local neurogenic as well as endothelial function is unaffected. PMID- 11918720 TI - Role of p38 MAPK in transforming growth factor beta stimulation of collagen production by scleroderma and healthy dermal fibroblasts. AB - Transforming growth factor beta has been implicated as a mediator of excessive extracellular matrix deposition in scar tissue and fibrosis, including systemic sclerosis. To further characterize the mechanism of collagen gene expression in systemic sclerosis and healthy skin fibroblasts, we examined the role of p38 MAPK signaling in collagen gene regulation by transforming growth factor beta. Treatment of dermal fibroblasts with transforming growth factor beta resulted in a prolonged activation of p38 MAPK. Furthermore, a specific inhibitor of p38 suppressed transforming growth factor beta stimulation of collagen type I mRNA and the alpha2(I) collagen promoter activity. To further probe the role of p38 in collagen regulation by transforming growth factor beta, we utilized an expression vector containing p38alpha cDNA. Ectopic expression of p38alpha enhanced COL1A2 promoter activity and potentiated transforming growth factor beta stimulation of this promoter. The p38 response element in the COL1A2 promoter overlapped with the previously characterized transforming growth factor beta response element. Consistent with these observations, collagen type I mRNA and protein levels were increased in transforming-growth-factor-beta-stimulated fibroblasts transduced with an adenoviral vector expressing p38alpha. To determine the possible role of p38 in abnormal collagen production by systemic sclerosis fibroblasts, p38 protein levels were compared in systemic sclerosis and healthy skin fibroblasts. Both cell types exhibited similar total levels of p38 MAPK and similar kinetics of p38 activation in response to transforming growth factor beta. In conclusion, this study demonstrates a costimulatory role for p38 MAPK in transforming growth factor beta induction of the collagen type I gene. Expression levels and activation status of p38 are not consistently elevated in systemic sclerosis fibroblasts suggesting that the p38 MAPK pathway is not dysregulated in systemic sclerosis fibroblasts. PMID- 11918722 TI - Hand eczema in Swedish adults - changes in prevalence between 1983 and 1996. AB - Hand eczema is the most frequent occupational skin disease. Our aim was to study changes in its prevalence in Swedish adults. Cross-sectional studies were performed in 1983 and 1996. Random samples from the population of Gothenburg, Sweden, aged 20-65 y, were drawn from the population register. Data were collected with a postal questionnaire, which was identical in the two studies. The response rate was 83.5% (16,708 out of 20,000) in 1983 and 73.9% (2218 out of 3000) in 1996. The reported 1 y prevalence of hand eczema decreased from 11.8% in 1983 to 9.7% in 1996 (p < 0.01), a large difference being found in the youngest age group. Reported childhood eczema increased from 10.4% to 12.4%, however (p < 0.01). Of those with childhood eczema 27.9% and 25.2% reported hand eczema, compared to 10.0% and 7.5% among those without childhood eczema. In total 76.8% were gainfully employed in 1983 and 68.3% in 1996 (p < 0.001). In 1983 23.0% were employed in "high-risk" occupations for hand eczema compared to 19.4% in 1996 (p < 0.001). Even though the increase in childhood eczema was largest in the youngest group, there was a large decrease in the prevalence of hand eczema in that age group among both sexes. The study indicates that the prevalence of hand eczema in Swedish adults had decreased between 1983 and 1996 despite an increasing prevalence of childhood eczema. Secular changes in reporting hand eczema and childhood eczema may explain some of the changes, but a decreased occupational exposure to skin irritants is a probable cause, implying that occupational factors may be important predictors of hand eczema. PMID- 11918721 TI - Inhibition of hair follicle growth by a laminin-1 G-domain peptide, RKRLQVQLSIRT, in an organ culture of isolated vibrissa rudiment. AB - We established a serum-free organ culture system of isolated single vibrissa rudiments taken from embryonic day 13 mice. This system allowed us to test more than 30 laminin-derived cell adhesive peptides to determine their roles on the growth and differentiation of vibrissa hair follicles. We found that the RKRLQVQLSIRT sequence (designated AG-73), which mapped to the LG-4 module of the laminin-alpha1 chain carboxyl-terminal G domain, perturbed the growth of hair follicles in vitro. AG-73 is one of the cell-binding peptides identified from more than 600 systematically synthesized 12 amino acid peptides covering the whole amino acid sequence of the laminin-alpha1, -beta1, and -gamma1 chains, by cell adhesion assay. Other cell-adhesive laminin peptides and a control scrambled peptide, LQQRRSVLRTKI, however, failed to show any significant effects on the growth of hair follicles. The AG-73 peptide binds to syndecan-1, a transmembrane heparan-sulfate proteoglycan. Syndecan-1 was expressed in both the mesenchymal condensation and the epithelial hair peg of developing vibrissa, suggesting that AG-73 binding to the cell surface syndecan-1 perturbed the epithelial-mesenchymal interactions of developing vibrissa. The formation of hair bulbs was aberrant in the explants treated with AG-73. In addition, impaired basement membrane formation, an abnormal cytoplasmic bleb formation, and an unusual basal formation of actin bundles were noted in the AG-73-treated-hair matrix epithelium, indicating that AG-73 binding perturbs various steps of epithelial morphogenesis, including the basement membrane remodeling. We also found a region-specific loss of the laminin-alpha1 chain in the basement membrane at the distal region of the invading hair follicle epithelium, indicating that laminins play a part in hair morphogenesis. PMID- 11918723 TI - A novel connexin 26 mutation in a patient diagnosed with keratitis-ichthyosis deafness syndrome. AB - Keratitis-ichthyosis-deafness syndrome is a rare disorder characterized by erythrokeratoderma, deafness, and keratitis. Scarring alopecia and squamous cell carcinoma can also occur. Most cases described so far were sporadic. Here we present evidence that keratitis-ichthyosis-deafness syndrome is caused by a mutation in the connexin 26 gene. This finding expands the spectrum of disorders caused by defects in connexin 26 and implies the gene in normal corneal function, hair growth, and carcinogenesis. PMID- 11918724 TI - Drug specific cytotoxic T-cells in the skin lesions of a patient with toxic epidermal necrolysis. AB - Toxic epidermal necrolysis is an extremely severe drug reaction, manifesting itself by widespread apoptosis of keratinocytes, generally considered to result from Fas/CD95-FasLigand interaction, but of unknown primary mechanism. We looked at the role of cells present in the skin blisters as probable effectors of this immune reaction. In a patient suffering from cotrimoxazole-induced toxic epidermal necrolysis, blister fluid cells were phenotyped by FACS and tested without prior restimulation for cytotoxicity on autologous and allogeneic cells in the presence of the drug. Blister fluid lymphocytes were predominantly CD8+, DR+, CLA+, CD56+ T lymphocytes, perforin positive and expressing preferentially two Vbeta chains of the T cell receptor repertoire. These lymphocytes were cytotoxic only in the presence of the drug towards autologous EBV transformed lymphocytes and towards allogeneic cells sharing HLA-Cw4. Cytotoxicity occurred in the presence of either cotrimoxazole, sulfamethoxazole, or the nitroso metabolite of sulfamethoxazole, but not with the hydroxylamine metabolite of sulfamethoxazole. The lysis was blocked by an anti-MHC class I monoclonal antibody. It was abolished by EGTA and CMA, but neither by anti-fas, brefeldin A, nor by anti-TRAIL receptor monoclonal antibodies, strongly suggesting perforin/granzyme-mediated cytotoxicity, without implication of Fas or TRAIL at this stage. This is direct evidence that T lymphocytes present within the lesions of toxic epidermal necrolysis may exhibit, without any re-stimulation, a drug specific cytotoxicity against autologous cells. Harboring the markers of classical CTL and MHC class I restriction these lymphocytes reacted against the parent drug and one of its reactive metabolites. These results challenge several current concepts and could support new therapeutic approaches. PMID- 11918726 TI - Is folate a promising agent in the prevention and treatment of cardiovascular disease in patients with renal failure? AB - Management of the conventional cardiovascular risk factors is insufficient to prevent the dramatic increase in atherosclerotic cardiovascular morbidity and mortality in patients with renal failure. Folate recently received attention as a potential alternative treatment option to decrease the excess cardiovascular risk in the uremic population. Folate administration is the principal treatment modality for hyperhomocysteinemia. Hyperhomocysteinemia is prevalent in more than 85% of patients with end-stage renal disease (ESRD) and is independently associated with increased odds for atherosclerotic cardiovascular disease. Several attempts have been made to normalize homocysteine levels in uremic patients with folate-based vitamin regimens. Although supraphysiologic doses of folic acid afford greater reductions in homocysteine levels than standard doses, the response to treatment is generally only partial and the large majority of ESRD patients have residual hyperhomocysteinemia. Several defects in folate metabolism have been described in uremia, which may explain the relative folate resistance in patients with renal failure, but their clinical relevance remains uncertain. It appears unlikely that the hyperhomocysteinemia in ESRD can be cured solely with folic acid supplements, since folate does not affect the prolonged plasma elimination of homocysteine, which is the primary defect in homocysteine metabolism in uremia. Folate restores endothelial dysfunction, associated with hyperlipidemia, diabetes and hyperhomocysteinemia. The beneficial effect appears to be independent of its homocysteine-lowering capacity and is possibly related to an improved bioavailability of nitric oxide. However, folate has failed to improve endothelial dysfunction in uremic patients. In the ESRD population, multiple metabolic and hemodynamic abnormalities adversely affect endothelial function. In addition, irreversible structural vascular disease already may be present. Folate should, therefore, probably be an integral part of an "endothelial protective regimen," consisting of lipid-lowering agents, antihypertensives and antioxidant vitamins and started very early in patients with renal failure. Before large-scale folate administration can be recommended, effects on hard endpoints of cardiovascular disease need to be demonstrated in randomized trials. Such trials are currently underway in patients with normal renal function at high risk for cardiovascular disease, and one trial has recently been initiated in stable renal transplant recipients. PMID- 11918727 TI - Pharmacology of cyclooxygenase-2 inhibition in the kidney. AB - Cyclooxygenase (COX) exists as two unique isoforms (that is, COX-1 and COX-2) which are poorly understood with regard to their roles in renal function. The renal effects of conventional non-steroidal anti-inflammatory drugs (NSAIDs) are believed to result from the inhibition of one or both isoforms. Drugs that selectively inhibit COX-2 provide useful pharmacological tools for discerning the effects associated with the inhibition of the individual isoforms, and may help clarify the renal roles of COX-1 and COX-2. This review summarizes the current data on the renal expression of COX isoforms and their potential roles in renal function, and reviews the studies that have attempted to correlate renal functional changes with selective isoform inhibition. Since there are significant differences in the expression of COX isoforms in the kidneys of laboratory animals and humans, this review also examines the correlation of the results of COX inhibition in experimental studies in laboratory animals with clinical data. Because of potential interspecies differences in the roles of COX isoforms in renal function, animal models may have limited predictive value for patients, particularly those with renal risk factors. Accordingly, any uncertainty concerning the safety or therapeutic benefit of COX-2-specific drugs in these patient populations will need to be resolved with clinical investigations. PMID- 11918728 TI - Caspases, Bcl-2 proteins and apoptosis in autosomal-dominant polycystic kidney disease. AB - BACKGROUND: Apoptosis is a characteristic feature of human autosomal-dominant polycystic kidney disease (ADPKD). The Han:Sprague-Dawley (SPRD) rat model closely resembles human ADPKD and presents an opportunity to investigate the apoptotic pathway in the pathogenesis of this disease. METHODS: Han:SPRD rats were studied during the early stages of ADPKD (newborn, 2 and 6 weeks old). Apoptotic cells were detected by the TUNEL (Tdt-mediated dUTP nick end-labeling) assay. Caspase-3 activity was measured using the fluorescent substrate DEVD-AMC and cleavage of poly (ADP-ribose) polymerase [PARP]. Expression of pro- and anti apoptotic B-cell lymphoma (Bcl-2) proteins was detected on Western blot analysis. RESULTS: TUNEL (+) cells, caspase-3 activity and caspase-mediated PARP breakdown were significantly increased in 2-week-old heterozygous (Cy/+) and homozygous (Cy/Cy) rat kidneys compared to normal littermate controls. In Cy/+ rat kidneys, decreased expression of anti-apoptotic Bcl-XL coincided with increased caspase-3 activity at 2 weeks of age while expression of Bcl-2, another anti-apoptotic protein, increased at 6 weeks of age. In Cy/Cy rat kidneys, decreased expression of Bcl-XL and increased expression of Bcl-2 was present at 2 weeks of age. Pro apoptotic Bax and Bad expression was unchanged at 2 weeks of age in both Cy/+ and Cy/Cy rat kidneys. CONCLUSIONS: Activation of caspase-3 and dysregulation of the balance between pro- and anti-apoptotic Bcl-2 family members, specifically a down regulation of anti-apoptotic Bcl-XL, correlates with increased apoptosis in polycystic Han:SPRD rat kidneys. PMID- 11918729 TI - Protein restriction in pregnancy is associated with increased apoptosis of mesenchymal cells at the start of rat metanephrogenesis. AB - BACKGROUND: In rats, offspring born to mothers supplied low protein diets during pregnancy have fewer glomeruli than normal. We hypothesized that such nephron deficits are associated with altered cell turnover in the metanephros, the embryonic precursor of the adult kidney. METHODS: Wistar rats were supplied with one of three isocaloric diets from day 0 of pregnancy: control (18% protein) or low protein (9% or 6%) diets. All had a normal chow after birth. Groups were compared by multilevel statistical modeling. RESULTS: At two weeks postnatally, when nephrogenesis has finished, controls had 16.8 x 103 +/- 0.7 x 10(3) (mean +/ SEM) glomeruli/kidney, whereas offspring exposed to 9% diet had 5.1 x 10(3) +/- 1.2 x 10(3) fewer and those exposed to 6% diet had 6.9 x 10(3) +/- 1.7 x 10(3) fewer glomeruli/kidney (P < 0.001, both diets). At embryonic day 13 (E13), when the metanephros has just formed, control metanephroi contained 2.35 x 10(4) +/- 0.15 x 10(4) cells, with no significant differences in low protein groups. At E15, when mesenchyme begins forming primitive nephrons but glomeruli are still absent, controls had 2.00 x 10(6) +/- 0.13 x 10(6) cells. E15 embryos exposed to 9% protein had 1.09 x 10(6) +/- 0.36 x 10(6) fewer cells/metanephros than controls, while those exposed to 6% diet had 1.45 x 10(6) +/- 0.37 x 10(6) fewer (P < 0.01, both diets). Apoptotic cells were detected by molecular (in-situ end labeling) and morphological (propidium iodide staining) techniques. In all diets, apoptosis was noted in condensing mesenchyme (nephron precursors) and loose mesenchyme (interstitial precursors). Control E13 metanephroi had 63 +/- 7 apoptotic cells/mm2, whereas those exposed to 9% diet had an increase of 77 +/- 26 cells/mm2 (P < 0.01) and those exposed to 6% diet had an increase of 55 +/- 26 cells/mm2 (P < 0.05). By E15, apoptosis was similar in all groups but metanephric mitosis was significantly increased in the 6% protein diet group. No change was found in the level of apoptosis in E13 mesonephroi. CONCLUSIONS: Maternal low protein diets reduce final numbers of glomeruli in association with enhanced deletion of mesenchymal cells at the start of kidney development. Whether aberrant nephrogenesis is a direct effect from deletion of nephron precursors, or an indirect effect from loss of supportive interstitial precursors, requires further investigation. PMID- 11918730 TI - Solitary functioning kidney and diverse genital tract malformations associated with hepatocyte nuclear factor-1beta mutations. AB - BACKGROUND: Renal tract malformations are, on occasion, associated with uterine malformations. The transcription factor hepatocyte nuclear factor (HNF)-1beta is expressed from the earliest stages of development of the Wolffian duct, the mesonephros and metanephros, and the Mullerian ducts in the mouse. In adult mice HNF-1beta is expressed in the kidney tubules, collecting ducts, and in the oviducts and uterus in the female (Mullerian duct derivatives) and in the epididymis, vas deferens and seminal vesicles (Wolffian duct derivatives) in the male. HNF-1beta mutations have been reported in two families where affected members have renal abnormalities, female genital tract malformations and early onset diabetes. Renal and uterine abnormalities have not been described in families without early-onset diabetes. METHODS: We sequenced the HNF-1beta gene in nine subjects with renal abnormalities and a personal or family history of female genital tract malformations, but no history of diabetes. RESULTS: Two families were identified with novel HNF-1beta mutations: a missense mutation in exon 2 with conversion of serine to proline at codon 151 (S151P) and a frameshift mutation in exon 3 with a 1 base pair deletion at codon 243 (Q243fsdelC). The S151P mutation proband has cystic kidneys and uterus didelphys. Her affected second son has renal cysts and hypospadias. The Q243fsdelC proband has a single functioning kidney and her two children have renal dysplasia. Histology in one child shows cystic dysplasia with a lack of glomeruli. The proband's sister is a mutation carrier and has a bicornuate uterus. Diabetes is not a feature in either family. CONCLUSIONS: This study confirms an association between HNF-1beta mutations and renal and Mullerian anomalies. The hypospadias may be coincidental. This study describes the first HNF-1beta mutations that are associated with a single functioning kidney and the absence of diabetes. This study further reinforces the variability of the renal and non-renal phenotypes associated with HNF-1beta mutations. PMID- 11918731 TI - ERK and p38 MAP kinase are required for rat renal development. AB - BACKGROUND: We previously demonstrated that extracellular signal-regulated protein kinase (ERK) and p38 mitogen-activated protein (MAP) kinase (p38) are strongly expressed in the embryonic kidney. In the present study, we investigated the role of ERK and p38 during kidney development. METHODS: Rat metanephroi were cultured from 15-day-old embryos, and exposed to inhibitors of MEK, an activator of ERK, PD98059 (300 micromol/L), U0126 (10 micromol/L), or a p38 inhibitor SB203580 (30 micromol/L) 24 to 120 hours after the start of culture. Growth of metanephroi was measured by surface area and thymidine incorporation. Ureteric buds and glomeruli were identified by labeling with Dolichos biflorus lectin and peanut agglutinin, respectively. PCNA staining and TUNEL assay were performed on kidney sections. The level of apoptosis was evaluated by examining DNA ladder formation. RESULTS: Growth of metanephroi was significantly inhibited by SB203580 but not by PD98059 or U0126. Ureteric bud branching was not affected by SB203580 or MEK inhibitors. Glomerular number was markedly reduced by SB203580 and to a lesser extent by U0126 (14 +/- 2 and 48 +/- 10% of controls, respectively). On histological examination, the number of tubuloglomerular structures was reduced in MEK inhibitor-treated metanephroi compared to controls. Very few mesenchymal condensates were observed in kidneys incubated with SB203580. PCNA-positive cells were reduced in SB203580-treated metanephroi compared to control and PD98059 treated kidneys. Apoptosis was increased in SB203580-treated kidneys and to a lesser extent in PD98059-treated cultures. CONCLUSIONS: Both ERK and p38 are required for renal development. ERK appears to play a role in nephrogenesis and p38 for kidney growth and nephrogenesis. PMID- 11918732 TI - Altered primate glomerular development due to in utero urinary tract obstruction. AB - BACKGROUND: In utero urinary tract obstruction is an important cause of newborn and childhood renal failure. Ureteric obstruction during active nephrogenesis results in cystic renal dysplasia; the earlier and longer the obstruction the more severe the histopathological changes of dysplasia. We have reported on a non human primate model of non-surgical in utero fetal ureteric obstruction that accurately reflects the human equivalent of obstructive renal dysplasia. A striking feature of this model is the effect of obstruction on normal glomerular development and podocyte survival. METHODS: To study the effect of urinary obstruction on glomerular development, kidneys were studied from fetuses undergoing unilateral ureteric obstruction by ultrasound guided injection of alginate beads as early as 75 days gestation (term gestation = 165 +/- 10 days). These kidneys displayed all the features of human obstructive cystic dysplasia, had reduced weights, and significant deficiencies in terminal ureteric duct branching. RESULTS: A combination of histochemistry, histomorphometry, and immunocytochemistry was used to demonstrate deficient cortical ureteric duct development and branching, reduced glomerular number, and altered glomerular basement membrane formation with in utero urinary tract obstruction. CONCLUSIONS: These data suggest that urinary tract obstruction during active nephrogenesis results in a defect in ureteric duct branching morphogenesis, and altered vascularization of the glomerulus with consequent podocyte dropout and decreased glomerular number. These abnormalities reflect human renal dysplasia, which is associated with compromised postnatal renal function and, thus, should be predictive of postnatal outcome. PMID- 11918733 TI - Role of the alpha-adducin genotype on renal disease progression. AB - BACKGROUND: A genetic susceptibility to hypertension may predispose to the development of end-stage renal disease (ESRD) and promote a more rapid loss of renal function in patients with renal diseases. The alpha-adducin (ADD) gene, alone or in combination with the angiotensinogen (AGT) and the angiotensin converting enzyme (ACE), is a candidate for abnormal blood pressure regulation and thus for increased susceptibility or faster progression to ESRD. METHODS: Genotyping for the G460W-ADD, M235T-AGT and the insertion/deletion (I/D)-ACE gene polymorphisms was performed in 260 control subjects and 260 ESRD patients using polymerase chain reaction, gel analysis and appropriate restriction digest. RESULTS: The frequencies of the ADD, AGT and ACE genotypes in ESRD patients did not differ from observed frequencies in control subjects. The average (+/-SE) time from diagnosis to the onset of ESRD tended to be shorter in the presence of the ADD-460WW (5.1 +/- 1.1 years, N = 10) than with the GW (9.9 +/- 0.7 years, N = 81) and GG (11.3 +/- 1.0 years, N = 164) genotypes (F-ratio=2.71, P = 0.068; WW vs. GW P < 0.06 and vs. GG <0.03). In the 167 patients homozygous for the ADD-G allele, a more rapid progression with the ACE-DD genotype as compared to ACE-DI and II was found (P < 0.02). CONCLUSIONS: The ADD genotype is predictive of the course of renal function loss in an unselected renal population and influences the effect of the ACE genotype to modulate the rate of progression to ESRD. Thus, the ADD genotype may play a role for the understanding of interindividual differences in the course of renal diseases. PMID- 11918734 TI - Adenosine-induced apoptosis in glomerular mesangial cells. AB - BACKGROUND: Mesangial cell apoptosis is a mechanism of resolution of glomerular hypercellularity in inflammatory forms of glomerular injury in which adenosine (ADO) was shown to play an anti-inflammatory role. This, and the observation that mesangial cell have ADO receptors prompted us to determine whether ADO induces mesangial cell apoptosis and to explore underlying mechanisms. METHODS: Cultured mouse mesangial cell were incubated in the presence or absence of ADO or ADO receptor agonists (R-PIA, NECA, IB-MECA, CGS26180) or antagonists (DPCPX, DPSPX, MRS1191) for 48 hours. Cell death was assessed by trypan blue exclusion analysis. Apoptosis was assessed by DNA fragmentation, TUNEL staining and flow cytometry. RESULTS: ADO and the A3 ADO receptor agonist IB-MECA induced mesangial cell death, which was markedly attenuated by the A3 receptor antagonist MRS1191. The A1 receptor agonist R-PIA, A2 receptor agonist NECA or the A2a receptor agonist CGS-12680 had no effect. The IB-MECA-induced mesangial cell death was due to apoptosis. This occurred via a cAMP independent mechanism. RT-PCR analysis revealed presence of A3, A1 and A2b but lack of A2a receptor transcripts in MC total RNA. Western blot analysis of mesangial cell lysates revealed expression the A3 receptor protein only. CONCLUSION: The observations indicate that ADO induces mesangial cell apoptosis via stimulation of the A3 receptor. PMID- 11918735 TI - Nutrition in CAPD: serum bicarbonate and the ubiquitin-proteasome system in muscle. AB - BACKGROUND: Metabolic acidosis in chronic renal failure (CRF) induces loss of lean body mass while elimination of acidosis during a one year trial improved anthropometric indices in continuous ambulatory peritoneal dialysis (CAPD) patients. In rats with CRF, the mechanisms causing loss of lean body mass have been linked to acidosis-induced destruction of the essential, branched-chain amino acids (BCAA) and activation of the ubiquitin-proteasome system that degrades muscle protein; the latter response includes increased transcription of the ubiquitin gene. METHOD: Our aim was to determine if increasing the serum bicarbonate (HCO3) concentration of CAPD patients would improve their nutritional status, increase plasma BCAA levels, and reduce ubiquitin mRNA in their muscle as an index of suppressed activity of the ubiquitin-proteasome system. Eight, stable, long-term CAPD patients underwent vastus lateralis muscle biopsy before being randomized to continue 35 mmol/L lactate dialysate or convert to a 40 mmol/L lactate dialysate. After four weeks, measurements were repeated. RESULTS: Serum HCO3 increased in all patients and final values did not differ statistically between the two groups so results for all patients were combined. Weight and body mass index increased significantly as did plasma BCAA. Muscle levels of ubiquitin mRNA decreased significantly; serum tumor necrosis factor alpha (TNF-alpha) also decreased. CONCLUSION: Our results indicate that even a small correction of serum HCO3 improves nutritional status, and provide evidence for down-regulation of BCAA degradation and muscle proteolysis via the ubiquitin proteasome system. Whether acidosis and inflammatory cytokines (such as, TNF alpha) interact to impair nutrition is unknown. PMID- 11918736 TI - 15-Deoxy-Delta12,14-prostaglandin J2 regulates mesangial cell proliferation and death. AB - BACKGROUND: Proliferation of intrinsic glomerular cells is a common response to renal injury. Acutely, proliferation may be beneficial, but sustained glomerular hypercellularity after injury is associated with progressive renal failure. To identify endogenous factors that may be responsible for regulating glomerular cell number, the effects of J-series cyclopentenone prostaglandins (PGs) on human glomerular mesangial cell proliferation and death were examined. METHODS: Human mesangial cells were grown in the presence or absence of PGJ2 or its metabolite 15-Deoxy-Delta12,14-PGJ2 (15dPGJ2). The number of viable cells was measured by the reduction of the tetrazolium MTS to a colored formazan product. Apoptosis was assessed by caspase-3 activation and DNA fragmentation. RESULTS: PGJ2 at concentrations up to 10 micromol/L caused mesangial proliferation. 15dPGJ2 also caused mesangial proliferation at low concentrations (< or =2.5 micromol/L), but induced mesangial cell death at higher concentrations (>5 micromol/L). Cell death occurred in part through apoptosis, measured as an increase in caspase-3 activity and DNA fragmentation in 15dPGJ2-treated cells. Cell death was associated with a decline in baseline phosphorylation of the survival factor Akt and increased Akt degradation, whereas 15dPGJ2-induced mesangial proliferation was blocked by inhibition of the PI 3-kinase/Akt pathway. 15dPGJ2 is a potent PPARgamma agonist. Like 15dPGJ2, treatment of mesangial cells with thiazolidinedione-type PPARgamma ligands (10 to 20 micromol/L) caused significant cell death, but at lower concentrations also caused a small degree of proliferation. CONCLUSIONS: J-series prostaglandins thus may be involved in the initiation of glomerular hypercellularity through Akt-dependent proliferation, and restoration of normal glomerular architecture through PPARgamma-mediated apoptosis. Manipulation of these prostaglandins may be relevant to the treatment of progressive glomerular disease. PMID- 11918737 TI - Transfection of tubule cells with Fas ligand causes leukocyte apoptosis. AB - BACKGROUND: Since the Fas/Fas Ligand (FasL) interaction is recognized as a major pathway of apoptosis in immune cells, we hypothesized that selective expression of FasL by tubular cells (TC) may promote the resolution of interstitial inflammation by inducing apoptosis of infiltrating immune cells. In this study, the effect of FasL transfection of rat TC on apoptosis of leukocytes was examined. METHODS: Rat tubule cells (NRK52E) were transfected with plasmids constructed using human and rat FasL (hFasL and rFasL). The propensity of activated, transfected TC to undergo apoptosis was examined. Similarly, the effects of FasL transfection on apoptosis of Jurkat cells and activated leukocytes were assessed directly following co-culture for 12 hours and in a cell insert system intended to assess the effects of soluble FasL. Fas and FasL expression was assessed by flow cytometry and apoptosis was examined using Annexin V staining and the TUNEL method. RESULTS: Expression of FasL in TC was increased after FasL transfection. Transfected TC showed no detectable increase in apoptosis following lipopolysaccharide (LPS) or tumor necrosis factor-alpha (TNF-alpha) activation. Jurkat cell apoptosis was increased ninefold and eightfold after co-culture with TC transfected with hFasL and rFasL, respectively (67.0 +/- 12.1% and 60.1 +/- 8.8% vs. 6.7 +/- 1.8% with un-transfected TC, P < 0.01). Similarly, apoptosis of activated leukocytes was increased fourfold by co culture (26.8 +/- 4.9% vs. 6.7 +/- 2.0% with untransfected TC, P < 0.01). Leukocyte apoptosis also was increased in an insert culture system (18.2 +/- 4.4% vs. 5.8 +/- 2.3% with un- transfected TC, P < 0.01). No increase of TC apoptosis was detected in any of the co-culture experiments. CONCLUSION: Enhanced expression of FasL by TC is capable of inducing apoptosis of activated leukocytes, without evidence for increased susceptibility to apoptosis of the transfected cells themselves. This suggests a potential role for this approach in the limitation and resolution of renal tubulointerstitial inflammation. PMID- 11918738 TI - Insulin is a potent survival factor in mesangial cells: role of the PI3 kinase/Akt pathway. AB - BACKGROUND: Elucidating the mechanisms of apoptosis is important for understanding the molecular mechanisms underlying glomerular disease. The phosphatidylinositol 3 kinase (PI3-kinase)/Akt pathway is essential for survival signaling in non-renal cells. However, little is known about the anti-apoptotic effect of insulin and the role of the PI3-kinase/Akt pathway in mesangial cells (MC) apoptosis. METHODS: Apoptosis was induced in wild type, p27Kip1 (p27) -/- and p21Cip1/Waf1 (p21) -/- mouse MC by survival factor withdrawal, actinomycin D, ultraviolet (UV)-B irradiation and cycloheximide in the presence or absence of insulin (1 micromol/L) or insulin-like growth factor-I (IGF-I; 100 ng/mL). The activation and levels of Akt, extracellular signal regulated kinase (ERK) and specific cell cycle proteins were determined by Western blot analysis. RESULTS: Insulin and IGF-I inhibited wild-type MC apoptosis induced by survival factor withdrawal, actinomycin D, ultraviolet-B irradiation and cycloheximide and in p27 -/- MC when apoptosis was induced by survival factor withdrawal. Akt was activated by insulin and IGF-I during apoptosis. Blocking PI3-kinase with LY294002 reduced Akt activation and abrogated the anti-apoptotic effect of insulin. ERK was activated during apoptosis and blocking ERK activation with U0126 or PD98059 partially rescued MC from apoptosis. Moreover, insulin also suppressed ERK activation during apoptosis. Our results also showed that the CDK inhibitor p21 was increased by insulin and that p21 up-regulation was PI3 kinase/Akt pathway dependent. Furthermore, p21 -/- MC apoptosis induced by survival factor withdrawal was not rescued by insulin in contrast to the wild type and p27 -/- MC. These data suggest that p21 may have a critical role in the anti-apoptotic effect of insulin. CONCLUSIONS: Insulin is a potent survival factor for MC in response to a number of different apoptotic triggers, and this effect is mediated through the PI3-kinase/Akt pathway. Moreover, ERK and p21 may be involved in anti-apoptotic effect of insulin in MC. PMID- 11918739 TI - Treatment of a murine model of high-turnover renal osteodystrophy by exogenous BMP-7. AB - BACKGROUND: The secondary hyperparathyroidism of chronic kidney disease (CKD) produces a high turnover osteodystrophy that is associated with peritrabecular fibrosis. The nature of the cells involved in the development of peritrabecular fibrosis may represent osteoprogenitors expressing a fibroblastic phenotype that are retarded from progressing through osteoblast differentiation. METHODS: To test the hypothesis that osteoblast differentiation is retarded in secondary hyperparathyroidism due to CKD producing bone marrow fibrosis, we administered bone morphogenetic protein 7 (BMP-7), a physiologic regulator of osteoblast regulation, to C57BL6 mice that had CKD produced by electrocautery of one kidney followed by contralateral nephrectomy two weeks later. Following the second surgical procedure, a subgroup of mice received daily intraperitoneal injections of BMP-7 (10 microg/kg). Three to six weeks later, the animals were sacrificed, blood was obtained for measurements of blood urea nitrogen (BUN) and parathyroid hormone (PTH) levels, and the femora and tibiae were processed for histomorphometric analysis. RESULTS: The animals had significant renal insufficiency with BUN values of 77.79 +/- 22.68 mg/dL, and the level of renal impairment between the CKD untreated mice and the CKD mice treated with BMP-7 was the same in the two groups. PTH levels averaged 81.13 +/- 51.36 and 75.4 +/- 43.61 pg/mL in the CKD and BMP-7 treated groups, respectively. The animals with CKD developed significant peritrabecular fibrosis. In addition, there was an increase in osteoblast surface and osteoid accumulation as well as increased activation frequency and increased osteoclast surface consistent with high turnover renal osteodystrophy. Treatment with BMP-7 eliminated peritrabecular fibrosis, increased osteoblast number, osteoblast surface, mineralizing surface and single labeled surface. There was also a significant decrease in the eroded surface induced by treatment with BMP-7. CONCLUSIONS: These findings indicate that BMP-7 treatment in the setting of high turnover renal osteodystrophy prevents the development of peritrabecular fibrosis, affects the osteoblast phenotype and mineralizing surfaces, and decreases bone resorption. This is compatible with a role of osteoblast differentiation in the pathophysiology of osteitis fibrosa. PMID- 11918740 TI - Hyperplasia of the parathyroid gland without secondary hyperparathyroidism. AB - BACKGROUND: Low dietary phosphorus (P) prevents parathyroid gland (PTG) hyperplasia and the development of secondary hyperparathyroidism (SH) in uremic rats. The present study explores the effects of P restriction on parathyroid hormone (PTH) synthesis and secretion and PT cell growth in rats with established SH and PTG hyperplasia. METHODS: Normal and 5/6 nephrectomized rats were fed a high P (0.8%) diet. After two weeks, the normal rats and half of the uremic rats were sacrificed (U-HP) while the remaining uremic rats were switched to a low P (0.2%) diet (U-HP-LP). RESULTS: High dietary P induced a significant increase in serum P, PTH, and PTG weight, but not ionized calcium compared to normal animals fed the same diet (N-HP). P restriction returned serum P and PTH to normal levels by one week. In contrast, PTG size did not regress and glands remained enlarged for up to eight weeks with no evidence of apoptosis. Ribonuclease protection assay and metabolic labeling studies demonstrated similar PTH/actin mRNA ratios and 35S-labeled PTH among the three groups. Intracellular intact PTH was higher in U-HP and U-HP-LP rats compared to N-HP animals with no differences between the two uremic groups. PTG-PTH content correlated only with PTG weight, and serum PTH only with serum P. The PTG secretory response to calcium remained intact. CONCLUSIONS: In established chief-cell hyperplasia, P restriction restores normal serum PTH levels without affecting PTG hyperplasia, PTH synthesis, PTG cytosolic PTH or the PTH secretory response to calcium, suggesting an impaired exocytosis of PTH. PMID- 11918741 TI - FK506 ameliorates proteinuria and glomerular lesions induced by anti-Thy 1.1 monoclonal antibody 1-22-3. AB - BACKGROUND: We have previously reported that CD4 T lymphocytes and their cytokines contribute to development of Thy 1.1 glomerulonephritis (GN). FK506 is reported to suppress the production of Th1 cytokines. The aims of this study were to elucidate the role of Th1 cytokines on mesangial alteration and to examine whether FK506 is available for therapy of mesangial proliferative GN. METHODS: The effects of daily treatments of FK506 from day -5 and from day +1 of Thy 1.1 GN induction on glomerular alterations were analyzed. RESULTS: FK506 treatment with 1.0 and 0.3 mg/kg body weight (BW) daily from day 1 to day 4 significantly reduced the glomerular expression of mRNA for interferon-gamma (IFN-gamma; 1.0 mg/kg BW FK506, 32.4% to the placebo group, P < 0.01) and IL-2 (55.6%, P < 0.01) on day 5. FK506 treatment from day -5 of GN induction reduced proteinuria and glomerular alteration in a dose-dependent manner. Although no side effects were detected in rats with 0.3 mg/kg BW of FK506 treatment from day +1, the treatment also ameliorated proteinuria (day 14, 3.7 +/- 0.89 vs. 19.8 +/- 12.3 mg/100 g BW/day P < 0.05) and glomerular alterations [total cell number, 63.1 +/- 3.1 vs. 80.2 +/- 7.4, P < 0.01; matrix expansion, 0.90 +/- 0.30 vs. 1.34 +/- 0.27, P < 0.05; alpha-smooth muscle actin (alphaSMA) expression; 1.20 +/- 0.12 vs. 1.96 +/- 0.29, P < 0.01] on day 14. CONCLUSION: Th1 cytokines may play an important role in the development of mesangial proliferative glomerulonephritis, and could be targets for therapy. FK506 might be available for clinical use. PMID- 11918742 TI - NHE3 and NHERF are targeted to the basolateral membrane in proximal tubules of colchicine-treated rats. AB - BACKGROUND: Depolymerization of microtubules in proximal tubule (PT) cells of colchicine-treated rats causes disruption of vesicle recycling and redistribution of some brush-border membrane (BBM) transporters into cytoplasmic vesicles. NHE3, an isoform of the Na+/H+ exchanger in the PT cell BBM, is acutely regulated by a variety of mechanisms, including protein trafficking and interaction with the PDZ protein, NHERF. The effects of microtubule disruption by colchicine on NHE3 trafficking in PT and the potential role of NHERF in this process have not been studied. METHODS: Immunofluorescence and immunogold cytochemistry were performed on cryosections of kidney tissue, and immunoblotting of BBM isolated from the renal cortex and outer stripe of control and colchicine-treated (3.2 mg/kg, IP, a single dose 12 hours before sacrifice) rats. RESULTS: In cells of the convoluted PT (S1/S2 segments) of control rats, NHE3 was located mainly in the BBM; subapical endosomes were weakly stained. In cells of the straight PT (S3 segment), NHE3 was present in the BBM and in lysosomes. In colchicine-treated rats, there was a marked redistribution of NHE3 from the BBM into intracellular vesicles and the basolateral plasma membrane in the S1/S2 segments. In the S3 segment, the abundance of BBM NHE3 was not visibly changed, but NHE3-positive intracellular organelles largely disappeared, and the antigen was detectable in the basolateral plasma membrane. The PDZ protein NHERF followed a similar pattern: in control animals, it was strong in the BBM and negative in the basolateral membrane in cells along the PT. After colchicine treatment, expression of NHERF in the basolateral membrane strongly increased in all PT segments, where it colocalized with NHE3. CONCLUSIONS: The data indicate that: (a) microtubules are involved in the apical targeting of NHE3 and NHERF in renal PT cells, and (b) the parallel basolateral insertion of NHE3 and NHERF may represent an indirect targeting pathway that involves transient, microtubule independent basolateral insertion of these proteins, followed by microtubule dependent, vesicle-mediated transcytosis to the BBM. PMID- 11918743 TI - Troglitazone suppresses the secretion of type I collagen by mesangial cells in vitro. AB - BACKGROUND: Our laboratory has shown that troglitazone, a thiazolidinedione and peroxisomal proliferator activated receptor gamma (PPAR-gamma) agonist, prevents mesangial expansion and glomerulosclerosis in diabetic rats. We investigated and compared the action of two PPAR agonists at the level of the mesangial cell. METHODS: Rat mesangial cells were grown in medium containing 5 mmol/L glucose, 30 mmol/L glucose, or 5 mmol/L glucose plus 25 mmol/L mannitol. The cultures were either left untreated, treated with 10 micromol/L troglitazone (PPAR-gamma), or 100 micromol/L clofibrate (PPAR-alpha). The following parameters were used to assess mesangial cell responses: detection of PPAR-gamma and -alpha mRNA, the degree of PPAR-gamma and -alpha activation, spread cell area, total protein production, and laminin and type I collagen production. RESULTS: Reverse transcription-polymerase chain reaction (RT-PCR) showed the presence of PPAR gamma and -alpha mRNA in rat mesangial cells. PPAR-gamma and -alpha proteins are active in mesangial cells and the extent of activation is affected by different glycemic conditions. Troglitazone and clofibrate treatment corrected in part the increase in spread cell area seen in mesangial cells in hyperglycemic conditions. However, neither agonist corrected the increase in total protein production induced by hyperglycemia. Treatment with troglitazone resulted in a significant, specific decrease in type I collagen along with a slight decrease in laminin production in both medium conditions. Clofibrate had no effect on laminin synthesis in either medium condition but did decrease type I collagen synthesis in cells grown in hyperglycemic conditions. CONCLUSION: PPAR-alpha and -gamma mRNA signaling pathways are in place and active in mesangial cells. Both agonists affect the phenotypic behavior of mesangial cells and ameliorate changes resulting from hyperglycemia. The data indicate that the correction of mesangial cell phenotype by troglitazone may influence production/deposition of a pathological fibrotic connective tissue matrix (that is, type I collagen) by these cells. PMID- 11918744 TI - STAT1 and STAT3 mediate thrombin-induced expression of TIMP-1 in human glomerular mesangial cells. AB - BACKGROUND: Thrombin exhibits numerous biological effects on glomerular resident cells, such as cell proliferation, release and synthesis of cytokines and collagen, expressions of metalloproteinases and their inhibitors, especially tissue inhibitor of metalloproteinase-1 (TIMP-1). However, the signaling mechanisms underlying these cellular events have not been fully elucidated. The present study was designed to examine the role of signal transducers and activators of transcription (STAT) in thrombin-induced TIMP-1 expression in human mesangial cells. METHODS: Cultured human glomerular mesangial cells were incubated with thrombin up to 12 hours. The effects of the antisense of STAT1 and antisense of STAT3 on stimulated TIMP-1 mRNA levels and DNA-binding activities of both STAT1 and STAT3 were determined using Northern blot, electrophoretic mobility shift assay (EMSA), and supershift assay. RESULTS: Cultured human mesangial cells constitutively expressed TIMP-1, and thrombin induced TIMP-1 gene transcription in a time- and dose-dependent manner. Hirudin, a specific inhibitor of thrombin, could block thrombin-induced TIMP-1 expression. Thrombin also induced STAT-DNA binding activity in a similar time- and dose-dependent manner. In order to examine the role of STAT in thrombin-induced TIMP-1 expression, STAT1 and STAT3 antisense oligonucleotides were used. EMSA showed that STAT1 and STAT3 antisense oligonucleotides could inhibit both thrombin-induced STAT-DNA binding activities and TIMP-1 mRNA expression; the supershift assay showed that the SIF band consisted of STAT1 and STAT3 proteins. CONCLUSIONS: Both STAT1 and STAT3 may be involved, at least in part, in thrombin-induced expression of the TIMP-1 gene in cultured human mesangial cells. PMID- 11918745 TI - Protective effect of insulin on ischemic renal injury in diabetes mellitus. AB - BACKGROUND: An exceptional susceptibility to unilateral renal ischemia/reperfusion (I/R) injury resulting in inflammation, fibrosis, atrophy of the kidney, and end-stage renal disease (ESRD) has been demonstrated in the diabetic rat. The aim of this study was to examine whether insulin treatment would reduce I/R injury in diabetic kidneys. METHODS: Diabetes mellitus (DM) was induced in male Wistar rats by streptozotocin. I/R was achieved by clamping the left renal artery for 30 minutes. Treatment with long acting insulin was started 7 to 14 days before or one day after I/R. Short acting insulin was administrated 2 to 6 hours before the injury. Apoptosis was evaluated six hours after ischemia with the TUNEL-method. Four weeks after the clamping inulin clearance was measured and kidneys were removed for histopathological evaluation. RESULTS: In DM animals renal I/R caused massive induction of apoptosis in the renal medulla after six hours as well as inflammation, fibrosis, renal atrophy and anuria within four weeks. Treatment with long acting insulin before I/R resulted in decreased cell death and an almost complete protection of both renal function and histomorphology. Treatment with short acting insulin before I/R also decreased the loss of renal function. In contrast, insulin treatment after I/R did not protect the kidney from damage. CONCLUSIONS: This study shows that insulin treatment with a subsequent improved metabolic control before renal I/R protected kidneys from ESRD. PMID- 11918746 TI - Tubular and cellular localization of the cardiac L-type calcium channel in rat kidney. AB - BACKGROUND: The mRNAs of several types of calcium channels have been identified in intact rat kidney, and L-type calcium channels cause changes in intracellular calcium in primary cultures of distal tubule cells. The aim of this study was to evaluate the tubular and cellular distribution of the alpha1C subunit of the L type calcium channel in intact kidney. METHODS: RT-PCR and Northern blot analysis were used to assess the regional abundance of the mRNA of this channel. Immunocytochemistry combined with confocal microscopy and surface biotinylation were applied to determine the tubular and cellular localization of the protein. RESULTS: Northern blot and RT-PCR analysis indicated that the mRNA of the alpha1C subunit of the cardiac L-type calcium channel was present in whole rat kidney, kidney tubules and kidney cell lines. Western blot of lysates from whole kidney, kidney tubules or cell lines revealed bands of approximately 190 kD for the alpha1C subunit and approximately 60 kD for the beta3 subunit. Confocal immunohistochemistry indicated that the alpha1C subunit of this channel was co expressed in cells of the distal tubule that express calbindin-D28K, but not in intercalated cells. The alpha1C subunit was also highly expressed in both outer and inner medullary collecting ducts. Serial confocal microscopic images or surface biotinylation experiments determined that the channel was predominantly on the basolateral membrane but had some distribution on the apical membrane. CONCLUSIONS: The distribution and cellular localization of the alpha1C subunit of cardiac L-type calcium channel suggest it is probably involved in intracellular and membrane calcium signaling. PMID- 11918747 TI - Sorbitol transport in rat renal inner medullary interstitial cells. AB - BACKGROUND: Sorbitol plays an important role in renal osmoregulation. In the rat renal inner medulla sorbitol synthesis and sorbitol degradation are located in different cell types. Whereas sorbitol synthesis can be detected in the inner medullary collecting duct cells, sorbitol degradation takes place in the interstitial cells. Therefore, one can speculate that the cooperation between epithelial and interstitial cells requires sorbitol transport into interstitial cells. METHODS: Our studies were performed with an interstitial cell line derived from the renal inner medulla of Wistar rats. These cells have typical characteristics of renal fibroblasts. In addition, they possess a high activity of sorbitol dehydrogenase as determined in vivo. Uptake was measured by liquid scintillation counting. For studies on sorbitol metabolism sorbitol concentration was measured photometrically. RESULTS: The results show that sorbitol transport into interstitial cells occurs via a yet to be described transport system. No saturation of sorbitol transport could be found up to an extracellular sorbitol concentration of 80 mmol/L. The transport was neither sodium nor chloride dependent. Trans-stimulation increased the sorbitol uptake. Sorbitol uptake was less inhibited by cytochalasin B than 2-deoxy-D-glucose uptake. The transport showed a high affinity for sorbitol and only little inhibition of sorbitol uptake by substances with a similar structure was observed. CONCLUSIONS: Our results show a new sorbitol transport system in renal inner medullary interstitial cells, which is rather different from the described sorbitol permease in renal epithelial cells and from glucose transporters of the GLUT- and SGLT-family. PMID- 11918748 TI - Human uremic plasma increases microvascular permeability to water and proteins in vivo. AB - BACKGROUND: The risk of cardiovascular disease is significantly higher in patients with long-term uremia than in otherwise healthy adults. This is true even before patients proceed to dialysis, but the reason why cardiovascular risk is increased is unknown. Transvascular transport of lipids and other macromolecules in both large vessels and the microcirculation has been implicated in generation of cardiovascular disease. METHODS: To determine whether patients with long-term uremia have circulating factors that promote increased vascular permeability, we measured the effect of perfusing microvessels with uremic plasma in a non-mammalian model of vascular permeability measurement. RESULTS: Perfusion of frog mesenteric microvessels with dialyzed normal plasma did not result in an increase in either hydraulic conductivity (Lp, permeability of the vessel wall to water) or oncotic reflection coefficient (sigma, permeability to macromolecules, particularly proteins). Perfusion with dialyzed uremic plasma resulted in a very significant increase in vascular permeability to both water (Lp increased 8.8 fold from 4.1 to 36.4 x 10(-7) cm x s(-1) cm H2O(-1)) and proteins (sigma reduced from 0.93 to 0.53). CONCLUSIONS: These results suggest that one or more circulating macromolecules in uremic plasma are able to increase transvascular solute and fluid flux, and may underlie the increased cardiovascular risk found in these patients. PMID- 11918749 TI - L-arginine deficiency and supplementation in experimental acute renal failure and in human kidney transplantation. AB - BACKGROUND: The "L-arginine paradox" refers to situations where L-arginine (L Arg) supplementation stimulates nitric oxide (NO) synthesis, despite saturating intracellular concentrations. This paradox is frequently observed in acute renal failure (ARF). First, the effects of L-Arg on renal function of rats with ARF were studied. Based on the promising results from these initial studies, the second part of our study searched for a form of ARF in humans that could be studied easily under conditions with little variance and yet was linked with endothelial dysfunction. Thus, we investigated the effects of L-Arg supplementation immediately after kidney transplantation in 54 patients. METHODS: In uranyl nitrate-induced ARF in rats the effects of L-Arg and L-NNA (inhibitor of nitric oxide synthase; NOS) on glomerular filtration rate (GFR), renal plasma flow (RPF), blood pressure (BP) and NOx (NO2- +NO3-) excretion were examined. Tissue L-Arg levels, NOS activities, immunodetection of NOS and superoxide dismutase (SOD), activities of nicotinamide adenine dinucleotide phosphate (NADPH) oxidase and xanthine oxidase, and nitrotyrosine immunoreactive protein (NT-IR) were determined and compared to sham operated animals. Secondly, in a randomized, double-blind study, the effects of L-Arg on GFR and RPF were investigated in 54 kidney transplant recipients, receiving IV L-Arg for three days. GFR and RPF were measured on days 1, 3, 5 and 10 by scintigraphy. RESULTS: In experimental ARF, decreased RPF and GFR were associated with reduced tissue L Arg levels, endothelial NOS-III expression, NO formation and NOx excretion. Reduction in GFR, RPF and NOx excretion were reversed upon administration of exogenous L-Arg. There also was a loss of Cu,Zn-SOD, a key enzyme against oxidative stress, and an elevation of NT-IR, an indicator of nitrosative stress and suggested marker for pathological actions of NO. However, NT-IR was not dependent on de novo NO synthesis and not related to the functional effects of l Arg administration. In kidney transplant recipients receiving organs with a short cold ischemia time (CIT) and from young donors, that is, those with a higher likelihood of a functional endothelium, early administration of L-Arg improved renal function. CONCLUSION: Both experimental and clinical data show that ?L-Arg deficiency and endothelial dysfunction are pathomechanistically relevant in ARF. The data suggest a therapeutic potential for the administration of L-Arg in ARF and kidney transplantation, at least in patients receiving kidneys with shorter CIT and from younger donors. PMID- 11918750 TI - Long-term caffeine consumption exacerbates renal failure in obese, diabetic, ZSF1 (fa-fa(cp)) rats. AB - BACKGROUND: Our preliminary studies indicate that chronic caffeine consumption has adverse renal effects in nephropathy associated with high blood pressure and insulin resistance. The purpose of this study was to investigate the effects of early (beginning at 8 weeks of age) and long-term (30 weeks) caffeine treatment (0.1% solution) on renal function and structure in obese, diabetic ZSF1 rats. METHODS: Metabolic and renal function measurements were performed at six-week intervals and in a subset of animals (N = 6 per group) heart rate (HR) and mean arterial blood pressure (MABP) were monitored by a radiotelemetric technique. At the end of the study acute, measurements of renal hemodynamics and excretory function were conducted in anesthetized animals. RESULTS: Caffeine produced a very mild increase (4 to 5%) of MABP and HR, but greatly augmented proteinuria (P < 0.001), reduced creatinine clearance (P < 0.05) and had a mixed effect on metabolic status in obese ZSF1 rats. Caffeine significantly reduced body weight, glycosuria, fasting glucose and insulin levels and improved glucose tolerance, had no effect on elevated plasma triglycerides levels and significantly increased plasma cholesterol level (P < 0.001). Acute measurements of renal function revealed increased renal vascular resistance (95.1 +/- 11 vs. 50.7 +/- 2.4 mm Hg/mL/min/g kidney, P < 0.01) and decreased inulin clearance (0.37 +/- 0.11 vs. 0.97 +/- 0.13 mL/min/g kidney, P < 0.002) in caffeine-treated versus control animals, respectively. Caffeine potentiated the development of more severe tubulointerstitial changes (P < 0.05) and increased focal glomerulosclerosis (14.7 +/- 1.7 vs. 6.5 +/- 0.9%, caffeine vs. control, P < 0.002). CONCLUSION: The present study provides the first evidence that caffeine (despite improving insulin sensitivity) exacerbates renal failure in obese, diabetic ZSF1 rats. Further mechanistic studies of adverse renal effects of caffeine in chronic renal failure associated with metabolic syndrome are warranted. PMID- 11918751 TI - Microalbuminuria in hypertension is not a determinant of insulin resistance. AB - BACKGROUND: Microalbuminuria (MA) clusters with metabolic derangements linked to the insulin resistance syndrome, and is associated with increased risk of cardiovascular disease in both diabetes and hypertension. This study questioned if MA, reflecting endothelial damage, is directly linked to impaired insulin action. METHODS: MA was measured in two 24-hour urine samples in 84 persons with untreated hypertension recruited from a population survey (diastolic blood pressures 90 to 105 mm Hg). Thirty-one percent had MA values>20 microg/min (MA group, N = 26), and these were matched according to age, gender, and body-mass index with hypertensive persons without MA (non-MA group, N = 32) for comparison of the metabolic profile. Insulin sensitivity was measured with clamp techniques. RESULTS: The MA and non-MA groups were similar in their fasting and post-load glucose and insulin levels, in the first (930 +/- 594 vs. 1097 +/- 707 pmol/L) and second (1111 +/- 662 vs. 1163 +/- 702 pmol/L) phases of insulin release during a hyperglycemic clamp, and in their insulin sensitivity indices (0.16 +/- 0.10 vs. 0.17 +/- 0.13, P> 0.3 for all). The MA group had higher systolic blood pressure (157 +/- 13 vs. 150 +/- 12 mm Hg, P = 0.05) and a higher serum level of circulating advanced glycation end products (AGEs; 11.0 +/- 3.0 vs. 7.9 +/- 3.5 U/mL, P = 0.05) than the controls. No associations were found between MA and the insulin sensitivity index, or glucose and insulin levels. Weak associations were found with systolic blood pressure (r = 0.25, P = 0.05), AGEs (r = 0.27, P = 0.05), and smoking habits (r = 0.39, P = 0.01). CONCLUSION: In hypertension, MA is not a determinant of insulin resistance, provided confounding factors such as degree of adiposity are carefully controlled. PMID- 11918752 TI - Cystatin C is a more sensitive marker than creatinine for the estimation of GFR in type 2 diabetic patients. AB - BACKGROUND: Glomerular filtration rate (GFR) is the best overall index of renal function in health and disease. Inulin and 51Cr-EDTA plasma clearances are considered the gold standard methods for estimating GFR. Unfortunately, these methods require specialized technical personnel over a period of several hours and high costs. In clinical practice, serum creatinine is the most widely used index for the noninvasive assessment of GFR. Despite its specificity, serum creatinine demonstrates an inadequate sensitivity, particularly in the early stages of renal impairment. Recently, cystatin C, a low molecular mass plasma protein freely filtered through the glomerulus and almost completely reabsorbed and catabolized by tubular cells, has been proposed as a new and very sensitive serum marker of changes in GFR. This study was designed to test whether serum cystatin C can replace serum creatinine for the early assessment of nephropathy in patients with type 2 diabetes. METHODS: The study was performed on 52 Caucasian type 2 diabetic patients. Patients with an abnormal albumin excretion rate (AER) were carefully examined to rule out non-diabetic renal diseases by ultrasonography, urine bacteriology, microscopic urine analysis, and kidney biopsy. Serum creatinine, serum cystatin C, AER, serum lipids, and glycosylated hemoglobin (HbA1c) were measured. GFR was estimated by the plasma clearance of 51Cr-EDTA. In addition the Cockcroft and Gault formula (Cockcroft and Gault estimated GFR) was calculated. RESULTS: Cystatin C serum concentration progressively increased as GFR decreased. The overall relationship between the reciprocal cystatin C and GFR was significantly stronger (r = 0.84) than those between serum creatinine and GFR (r = 0.65) and between Cockcroft and Gault estimated GFR and GFR (r = 0.70). As GFR decreased from 120 to 20 mL/min/1.73 m2, cystatin C increased more significantly that serum creatinine, giving a stronger signal in comparison to that of creatinine over the range of the measured GFR. The maximum diagnostic accuracy of serum cystatin C (90%) was significantly better than those of serum creatinine (77%) and Cockcroft and Gault estimated GFR (85%) in discriminating between type 2 diabetic patients with normal GFR (>80 mL/min per 1.73 m2) and those with reduced GFR (<80 mL/min/1.73 m2). In particular, the cystatin C cut-off limit of 0.93 mg/L corresponded to a false positive rate of 7.7% and to a false-negative rate of 1.9%; the serum creatinine cut-off limit of 87.5 micromol/L corresponded to a false-positive rate of 5.8% and to a false-negative rate of 17.0%. CONCLUSIONS: Cystatin C may be considered as an alternative and more accurate serum marker than serum creatinine or the Cockcroft and Gault estimated GFR in discriminating type 2 diabetic patients with reduced GFR from those with normal GFR. PMID- 11918753 TI - Effects of enalapril and eprosartan on the renal vascular nitric oxide system in human essential hypertension. AB - BACKGROUND: Experimental data in humans on the contribution of angiotensin converting enzyme inhibitors and angiotensin II type 1 receptor blockers to the nitric oxide system of the renal vasculature are inconsistent. Enalapril and eprosartan, alone and in combination, were used to determine their short-term effects on the renal nitric oxide system and renal hemodynamics of human subjects with essential hypertension. METHODS: Twenty male, white patients (27 +/- 1 years) with mild essential hypertension (143 +/- 11/95 +/- 6 mm Hg) were included in a double-blind, randomized, placebo-controlled, fourfold cross-over study with placebo, enalapril (20 mg/day), eprosartan (600 mg/day), or combination of both drugs (10 and 300 mg/day, respectively) each over a one week period followed by a two-week washout phase. After each study phase the glomerular filtration rate (GFR) and renal plasma flow (RPF) were determined. Basal nitric oxide synthesis of the renal vasculature was assessed by the decrease in RPF after inhibition of nitric oxide synthase with NG-monomethyl-L-arginine (L-NMMA; 4.25 mg/kg). RESULTS: After one week of therapy, the combination therapy decreased casual blood pressure by 5 +/- 2/3 +/- 1 mm Hg versus placebo (P < 0.01). Neither enalapril alone (-2 +/- 2/1 +/- 2 mm Hg, NS vs. placebo) nor eprosartan alone (-1 +/- 1/0 +/- 2 mm Hg, NS vs. placebo) had a clear-cut significant effect on casual blood pressure. In the combination phase, RPF increased by 123 +/- 36 mL/min (P < 0.01). Neither enalapril alone (+59 +/- 46 mL/min, P = 0.21) nor eprosartan alone (+113 +/- 51 mL/min, P = 0.06) had a clear-cut significant effect on RPF. Changes of RPF induced by treatment correlated with the L-NMMA induced decrease in RPF in the combination (r = 0.70, P < 0.01) and eprosartan phase (r = 0.86, P < 0.001), but not in the enalapril phase (r = -0.44, P = 0.10). Renal vascular resistance was reduced by each active treatment with the most prominent reduction in the combination phase. GFR was unaffected by any treatment. CONCLUSIONS: In contrast to the effects of either substance alone, a combination of half the dose of eprosartan with half the dose of enalapril had a prominent effect on renal perfusion. The effects of eprosartan on RPF are mediated, at least in part, by an increased bioavailability of nitric oxide in the renal vasculature. PMID- 11918754 TI - A placebo-controlled trial examining atorvastatin in dyslipidemic patients undergoing CAPD. AB - BACKGROUND: Individuals with chronic renal disease are at high risk of cardiovascular morbidity and mortality, and therefore the management of dyslipidemia is particularly important in this patient population. This double blind randomized study investigated the efficacy and safety of the 3-hydroxy-3 methylglutaryl coenzyme A (HMG-CoA) reductase inhibitor, atorvastatin, in continuous ambulatory peritoneal dialysis (CAPD) patients with dyslipidemia. METHODS: Following a two- to four-week baseline period, patients with low-density lipoprotein (LDL)-cholesterol > or =3.5 mmol/L (135 mg/dL) were randomized to receive either atorvastatin 10 mg (N = 82) or placebo (N = 95) for 16 weeks. If LDL-cholesterol remained > or =3.5 mmol/L, the dose of atorvastatin was titrated to 20 mg and 40 mg after four and eight weeks, respectively. RESULTS: After four weeks a significantly greater proportion of patients receiving atorvastatin 10 mg had achieved the LDL-cholesterol goal < or =3.5 mmol/L compared with patients receiving placebo (85.4% vs. 16.0%; P < or = 0.001). The statistically significant difference between the two groups was maintained at week 8 and week 16 (P < or = 0.001 at both time points). At week 16, patients receiving atorvastatin had significantly greater reductions from baseline in LDL cholesterol, total cholesterol, triglycerides and total cholesterol:HDL cholesterol ratio (all P = 0.0001), and a significantly greater increase from baseline in HDL-cholesterol (P = 0.001) than patients receiving placebo. The overall adverse event profile for atorvastatin was similar to that observed with placebo. CONCLUSIONS: Atorvastatin was effective in achieving target LDL cholesterol levels in a high proportion of the dyslipidemic CAPD patients studied at doses that are well tolerated. PMID- 11918755 TI - Podocytopenia and disease severity in IgA nephropathy. AB - BACKGROUND: IgA nephropathy is a common form of progressive glomerular disease, associated with proliferation of mesangial cells and mesangial deposition of IgA. The present study was designed to investigate functional and morphological covariates of disease severity in patients with IgA nephropathy. METHODS: Glomerular hemodynamics, permselectivity and ultrastructure were studied in 17 adult patients with IgA nephropathy using inulin, para-aminohippuric acid (PAH) and 3H-Ficoll clearances and morphometric methods. A mathematical model of macromolecule permeation through a heteroporous membrane was used to characterize glomerular permselectivity. Controls consisted of 14 healthy living kidney donors and 12 healthy volunteers. RESULTS: The patients were heterogeneous in their disease severity, but as a group had a decreased glomerular filtration rate (GFR) and increased urinary protein excretion compared to controls [63 +/- 29 SD vs. 104 +/- 23 mL/min/1.73 m2, P < 0.001, and (median) 1.34 vs. 0.11 g/day, P < 0.0001, respectively). A multivariate analysis of structural and functional relationships revealed GFR depression to be most strongly correlated with the prevalence of global glomerular sclerosis (t = -4.073, P = 0.002). Those patients with the most severe glomerular dysfunction had a reduced number of glomerular visceral epithelial cells (podocytes) per glomerulus. The degree of podocytopenia was related to the extent of glomerular sclerosis and of impairment of permselectivity and GFR, with worsening injury below an apparent threshold podocyte number of about 250 cells per glomerulus. There were no corresponding correlations between these indices of injury and the number of mesangial and endothelial cells. CONCLUSIONS: Our findings show that podocyte loss is a concomitant of increasing disease severity in IgA nephropathy. This suggests that podocyte loss may either cause or contribute to the progressive proteinuria, glomerular sclerosis and filtration failure seen in this disorder. PMID- 11918756 TI - Moderate renal insufficiency and the risk of cardiovascular mortality: results from the NHANES I. AB - BACKGROUND: Conflicting evidence exists concerning whether renal insufficiency is an independent risk factor for cardiovascular disease in the general population. The objective of this study was to determine whether moderate renal insufficiency was associated with total and cardiovascular mortality, independent of traditional cardiovascular risk factors, in a community sample representative of the U.S. general non-institutionalized population. METHODS: Participants in the U.S. First National Health and Nutrition Examination Survey (NHANES I, 1974-1975) and NHANES I Epidemiologic Follow-up Study (NHEFS, 1992, 18 year follow-up) were evaluated. The primary analysis was limited to 2352 adults with complete data, and no baseline cardiovascular disease. A creatinine of 104 to 146 micromol/L in women, and 122 to 177 micromol/L in men (approximate glomerular filtration rate of 30 to 60 mL/min/1.73 m2) was defined as moderate renal insufficiency. Supplementary analyses included participants with marked renal impairment and baseline cardiovascular disease. RESULTS: The unadjusted hazard ratio for moderate renal insufficiency compared to preserved renal function was significant for total mortality (hazard ratio 1.7; 95% confidence interval 1.3 to 2.2), and for cardiovascular mortality (2.2; 1.5 to 3.1). After adjustment for traditional cardiovascular risk factors, there was no independent association between moderate renal insufficiency and total mortality (1.0; 0.8 to 1.4), or cardiovascular mortality (1.2; 0.8 to 1.8). These results were consistent in supplementary analyses. CONCLUSIONS: These results do not support moderate renal insufficiency as an independent risk factor for cardiovascular disease in the general population. The association between moderate renal insufficiency and cardiovascular disease, demonstrated in other epidemiologic studies, appears to be due to co-occurrence of renal insufficiency with traditional cardiovascular risk factors. PMID- 11918757 TI - Cyclophosphamide pharmacokinetics and dose requirements in patients with renal insufficiency. AB - BACKGROUND: Intravenous pulse administration of cyclophosphamide (CYC) has been successfully used for the treatment of various autoimmune diseases. These patients often present with impaired renal function or even end-stage renal failure. Nevertheless, data concerning pharmacokinetics of CYC in renal insufficiency (RI) and on hemodialysis (HD) are rare and contradictory. METHODS: The pharmacokinetics of CYC (0.5 to 1 g/m2 as a one-hour infusion) were determined in patients with renal involvement of autoimmune diseases. Group A (N = 6) patients had a creatinine clearance (CCr) of 25 to 50 mL/min, group B patients' (N = 5) CCr was 10 to 24 mL/min, and group C (N = 6) patients had CCr values <10 mL/min and HD. Concentrations of CYC in serum, dialysate and urine were measured by HPLC. Twelve previously investigated patients with normal renal function served as controls. RESULTS: Mean clearance (CL) of CYC was significantly reduced with decreased renal function (79 vs. 57 and 47 mL/min, controls vs. A and B, respectively, P < 0.05), but only moderately lower in the patients who received a three-hour HD during the study period (group C, 64 mL/min, NS). This resulted in reciprocal increases in systemic drug exposure (dose corrected AUC was 216, 298, 382 and 266 microg x h/mL x g, controls, A, B and C, respectively). Urinary excretion of CYC was markedly reduced in all patients with RI (renal CL was 14.9 vs. 3.4, 2.4 and 2.1 mL/min, controls vs. A, B and C, respectively, P < 0.001). However, in patient group C, a mean of 22% of administered CYC dose was eliminated by a three hour HD starting seven hours after CYC administration. Individual CCr values were significantly (P < 0.001) correlated with renal and systemic CL of CYC, respectively, and negatively correlated with dose corrected AUC. CONCLUSIONS: Clearance of CYC is decreased in patients with reduced renal function, thereby resulting in an increased systemic drug exposure. However, in hemodialysis-dependent patients, removal of CYC into the dialysate has to be taken into account. For optimal dosing of CYC in patients with renal insufficiency, the severity of renal impairment and the use and timing of hemodialysis have to be considered. PMID- 11918758 TI - Renal flares in 91 SLE patients with diffuse proliferative glomerulonephritis. AB - BACKGROUND: Even when treated with current protocols, 25 to 30% of systemic lupus erythematosus (SLE) patients with diffuse proliferative glomerulonephritis (DPGN) evolve to end-stage renal disease (ESRD). The occurrence of renal flares is considered to be an important risk factor for the evolution to ESRD. The aim of this retrospective study was to evaluate the incidence and prognostic significance of renal flares in SLE patients with DPGN and to identify predictors for the occurrence of flares. METHODS: Ninety-one SLE patients were selected for study based on the following criteria: (a) evidence of renal involvement, (b) a follow-up of at least 6 months after the renal biopsy, and (c) a steady improvement in renal manifestations after the biopsy lasting for at least three months. RESULTS: Renal flares occurred in 54% of the patients after renal biopsy and appropriate treatment. A younger age at the time of renal biopsy correlated with the occurrence of renal flares. A high activity index (> or =10) and karyorrhexis on histology correlated with the occurrence of nephritic flares. Twenty-seven percent of the patients developed ESRD. The number of renal flares, nephritic flares, and "early" proteinuric flares (that is, those occurring in the first 18 months after renal biopsy) as well as serum creatinine levels, karyorrhexis, and chronicity index on renal histology were correlated with doubling serum creatinine. CONCLUSIONS: Our results suggest that (a) a distinct subgroup of SLE patients exists, made up of younger patients with extensive, active lesions on renal biopsy, who are at higher risk for renal flares, (b) renal flares represent important predictors of doubling serum creatinine. PMID- 11918759 TI - HLA class I and II in black children with hepatitis B virus-associated membranous nephropathy. AB - BACKGROUND: The pathogenetic mechanisms by which individuals with chronic hepatitis B virus (HBV) infection develop membranous nephropathy (MN) are probably dependent on interactions between viral, host and environmental factors; some evidence suggests a genetic predisposition. HBVMN constitutes a major etiological group in black children with nephrotic syndrome. We therefore explored the HLA associations in black children with HBVMN. METHOD: Thirty black children, age range 2 to 16 years, with biopsy-proven HBVMN, were the subjects of the study. HBV status was determined using third generation ELISA. HLA A, B and C antigens were determined using a two-stage lymphocytotoxic test. HLA DRB1* and DQB1* typing was done using sequence-specific primers. HLA class 1 and II antigen frequencies of the study subjects were compared to controls that were randomly chosen healthy blood donors from the same population. RESULTS: HLA DQB1*0603 was increased in patients with HBVMN compared to controls (chi2 = 13.65, RR = 4.3). DRB1*07 and DQB1*02 were increased in frequency in the study subjects but failed to reach statistical significance. There was no significant difference in the frequencies of class 1 antigens in the study group compared to controls. CONCLUSION: To our knowledge, this is the first report of HLA associations in black patients with HBVMN in whom Class 1 and 11 antigens were determined using molecular methodology. There was a high frequency of DQB1*0603 in subjects compared to controls, suggesting a possible genetic predisposition to the development of HBVMN. PMID- 11918760 TI - Acute renal allograft rejection with intimal arteritis: histologic predictors of response to therapy and graft survival. AB - BACKGROUND: Acute renal allograft rejection with intimal arteritis is designated by the widely used Banff 97 classification as type 2A or 2B depending on the extent of arteritis, without regard to interstitial inflammation or tubulitis. We examined whether the distinction between type 2A and 2B is relevant to short- and long-term clinical outcomes, and if outcomes in this subset of acute rejection also are affected by tubulitis, interstitial inflammation, and several additional histologic and clinical parameters. METHODS: Pathology records were searched to identify cases of acute renal allograft rejection with intimal arteritis diagnosed between January 1985 and September 2000. For each case, the patient's chart was reviewed to determine the response of the rejection episode to therapy, type(s) of therapy given, and length of graft survival. All biopsies were reviewed and Banff acute and chronic indices recorded by a pathologist blinded to these data. Biopsies not showing type 2A or 2B rejection were excluded, as were repeat biopsies from the same patient and cases with recurrent glomerular disease, viral infection, donor-specific antibodies, or more than mild chronic change. RESULTS: The initial response to anti-rejection therapy was significantly worse in patients with type 2B acute rejection (N = 29) than in those with type 2A (N = 102) by univariate and multivariate analyses, despite more aggressive treatment of type 2B rejection. In a Cox proportional hazards model the hazard ratio for graft failure for 2B versus 2A was 1.9 (P = 0.05), but this was not significant when adjusted for the initial response to therapy. Cases with minimal or mild tubulitis responded better to therapy than those with moderate or severe tubulitis, although graft survival was not significantly affected by the tubulitis score. CONCLUSIONS: The distinction between types 2A and 2B acute rejection in the Banff 97 classification has significant prognostic value with regard to both short- and long-term clinical outcomes, although the difference in long-term graft survival is mainly related to the initial response to therapy. Reports of biopsies showing type 2A or 2B rejection also should specify the degree of tubulitis present, as the latter may significantly influence the initial response to therapy. PMID- 11918761 TI - Effects of pH-neutral, bicarbonate-buffered dialysis fluid on peritoneal transport kinetics in children. AB - BACKGROUND: Due to their superior biocompatibility, pH-neutral solutions are beginning to replace acidic lactate-buffered peritoneal dialysis (PD) fluids. We hypothesized that pH-neutral and acidic solutions might differentially affect peritoneal transport in the early dwell phase, due to differences in ionic shifts and initial peritoneal vasodilation. Such differences may become clinically relevant in patients with frequent short cycles on automated PD (APD). METHODS: Twenty-five children were treated with a lactate-buffered (35 mmol/L, pH 5.5) or a bicarbonate-buffered PD solution (34 mmol/L, pH 7.4) in randomized order on two sequential days. Each day a four-hour Standardized Permeability Analysis (SPA) was performed, followed by overnight APD (7 cycles, fill volume 1000 mL/m2, dwell time 75 min). Functional peritoneal surface area was dynamically assessed using the three-pore model. RESULTS: While intraperitoneal pH was constant at 7.41 +/- 0.03 throughout the SPA with bicarbonate fluid, the dialysate remained acidic for more than one hour with lactate solution (pH 7.12 +/- 0.08 at 1 h). Total pore area was 60% higher during the first 30 minutes of the dwell than under steady state conditions, without a difference between acidic and pH-neutral fluid. Net base gain, intraperitoneal volume kinetics, glucose absorption, ultrafiltration rate, effective lymphatic absorption and the transport of urea, potassium, beta2 microglobulin and albumin were similar with both fluids. However, phosphate and creatinine elimination were 10% lower with bicarbonate PD fluid, resulting in corresponding significant decreases in the 24-hour clearances of these solutes. CONCLUSION: The peritoneal surface area is not measurably influenced by pH neutral PD fluid. Creatinine and phosphate elimination appears to be slightly reduced with bicarbonate fluid; this observation awaits clarification in extended therapeutical trials. PMID- 11918762 TI - Randomized prospective comparison between erythropoietin and androgens in CAPD patients. AB - BACKGROUND: Anemia and malnutrition are significant complications in peritoneal dialysis (PD) patients. Previous studies in hemodialysis have shown that androgens are effective as therapy for anemia; however, this has not been tested in a randomized prospective trial in PD patients. Furthermore, the anabolic properties of androgens may exert additional benefits on the nutritional status in this population. METHODS: Twenty-seven stable male patients over 50 years who were under maintenance continuous ambulatory peritoneal dialysis (CAPD) therapy were randomized to receive recombinant human erythropoietin (rHuEPO; N = 14) or nandrolone decanoate (ND; 200 mg/week IM; N = 13) as therapy for anemia. The evolution of hematologic parameters and the impact on both nutritional anthropometric and biochemical variables were evaluated after six months of treatment. RESULTS: Hemoglobin and hematocrit experienced similar increases in both groups: from 8.5 +/- 0.9 g/dL and 25.8 +/- 2.7% to 11.7 +/- 0.6 g/dL and 34.7 +/- 1.6% (P < 0.001) in patients receiving rHuEPO, and from 8.9 +/- 0.8 and 27 +/- 2.2% to 11.8 +/- 0.4 g/dL and 35.1 +/- 1.5% (P < 0.001) in subjects treated with ND. At the end of the study, out of the diverse nutritional variables included in this investigation, only weight and body mass index significantly increased in the rHuEPO group. Conversely, both anthropometric [weight, body mass index, triceps skinfold, mid-arm circumference (MAC) and mid arm muscle circumference (MAMC)] and biochemical parameters (serum total proteins, albumin, prealbumin and transferrin) were significantly increased in patients treated with ND. In this group, serum urea nitrogen, urea net excretion and protein equivalent of nitrogen appearance significantly decreased. These facts, together with an increase in serum creatinine and no changes in dietary intake during the study, suggest a rise in muscle mass related to an anabolic effect of nandrolone decanoate. Interestingly, serum levels of insulin-like growth factor type 1 (IGF-1) increased in patients on the androgen group compared to subjects treated with rHuEPO. Moreover, there was a positive and significant correlation between the rise in IGF-1 concentrations and the increase in hemoglobin, hematocrit, MAC and MAMC. CONCLUSIONS: Androgens therapy improved the anemia in elderly male CAPD patients in a similar manner to that observed with rHuEPO. Furthermore, compared with rHuEPO, androgen administration was associated with beneficial effects on nutritional status. The mechanism of action of androgens on hematologic and nutritional parameters might be mediated, at least in part, by IGF-1. PMID- 11918763 TI - ACE inhibitors and AT1 receptor antagonists: is two better than one? PMID- 11918764 TI - Serum cystatin C in transplantation. PMID- 11918765 TI - Continuous renal replacement therapy versus intermittent hemodialysis in acute renal failure. PMID- 11918767 TI - Fragmentation of filtered proteins and implications for glomerular protein sieving in Fanconi syndrome. PMID- 11918768 TI - Red blood cells and thin basement membrane disease. PMID- 11918769 TI - ANCA-associated renal vasculitis. PMID- 11918770 TI - Proteinuria in obstructive sleep apnea. PMID- 11918771 TI - The effect of fluid and sodium mortality on patients undergoing peritoneal dialysis. PMID- 11918772 TI - Acute inflammation in the pathogenesis of hemolytic-uremic syndrome. PMID- 11918773 TI - The contribution of haploids, diploids and clones to fine-scale population structure in the seaweed Cladophoropsis membranacea (Chlorophyta). AB - Local populations of Cladophoropsis membranacea exist as mats of coalesced thalli composed of free-living haploid and diploid plants including clonally reproduced plants of either phase. None of the phases are morphologically distinguishable. We used eight microsatellite loci to explore clonality and fine-scale patch structure in C. membranacea at six sites on the Canary Islands. Mats were always composites of many individuals; not single, large clones. Haploids outnumbered diploids at all sites (from 2:1 to 10:1). In both haploid and diploid plants, genetic diversity was high and there was no significant difference in allele frequencies. Significant heterozygote deficiencies were found in the diploid plants at five out of six sites and linkage disequilibrium was associated with the haploid phase at all sites. Short dispersal distances of gametes/spores and small effective population sizes associated with clonality probably contribute to inbreeding. Spatial autocorrelation analysis revealed that most clones were found within a radius of approximately 60 cm and rarely further than 5 m. Dominance of the haploid phase may reflect seasonal shifts in the relative frequencies of haploids and diploids, but may alternatively reflect superiority of locally adapted and competitively dominant, haploid clones; a strategy that is theoretically favoured in disturbed environments. Although sexual reproduction may be infrequent in C. membranacea, it is sufficient to maintain both life history phases and supports theoretical modelling studies that show that haploid diploid life histories are an evolutionarily stable strategy. PMID- 11918774 TI - Insertion polymorphism of retrotransposable elements in populations of the insular, endemic species Drosophila madeirensis. AB - The insertion site numbers of the retrotransposable elements (TE) 412, gypsy and bilbo were determined in individuals of five distinct natural populations of the endemic species Drosophila madeirensis from the island of Madeira. The TE distributions were compared to those of the paleartic, widespread and phylogenetically closely related species, D. subobscura. In situ hybridization and Southern blots showed that in D. madeirensis the number of insertion sites ranged between 10 and 15, three and six, and 35 and 42 for elements 412, gypsy and bilbo, respectively. The corresponding values for D. subobscura were similar. Two of these elements, 412 and gypsy, had very few insertions in the heterochromatin, unlike bilbo, which displayed a high heterochromatic insertion number. The Southern band polymorphism was very high, leading to within population variation of 97.2%, whatever the population and the TE concerned. Using the polymorphic TE insertion sites as markers to analyse population structure by AMOVA, adapted for RAPD (Randomly Amplified Polymorphic DNA) data, we found small but significant genetic differences between the populations on Madeira. This slight differentiation, coupled with similar copy numbers for each TE between populations, suggests that the D. madeirensis species consists of a single, only slightly subdivided population. These data also show that insular populations and endemic species of Drosophila can have as many copies of TEs as more widespread species. PMID- 11918775 TI - Genetic structure of natural populations of the grass endophyte Epichloe festucae in semiarid grasslands. AB - Plants of red fescue (Festuca rubra), a commercially important turf grass, are infected by the fungal endophyte Epichloe festucae in semiarid natural grasslands, known as dehesas, in western Spain. We used amplified fragment length polymorphism (AFLP) markers to analyse the genetic polymorphism existing in two natural populations of Epichloe festucae. Linkage disequilibrium and the presence of clonal lineages indicated that nonrecombinant asexual reproduction predominates in both populations. However, most genetic variation detected was found to occur within populations, with only a moderate amount of genetic differentiation between populations (F(ST): 0.197). Overall, the study suggests that dehesa grasslands are useful reservoirs of Epichloe festucae endophytes, and provides information on population structure which is relevant to design sampling strategies. PMID- 11918776 TI - Islands within an island: phylogeography and conservation genetics of the endangered Hawaiian tree snail Achatinella mustelina. AB - Mitochondrial DNA (mtDNA) sequences were used to evaluate phylogeographic structure within and among populations of three endangered Hawaiian tree snail species (n = 86). The primary focus of this investigation was on setting conservation priorities for Achatinella mustelina. Limited data sets for two additional endangered Hawaiian tree snails, A. livida and A. sowerbyana, were also developed for comparative purposes. Pairwise genetic distance matrices and phylogenetic trees were generated, and an analysis of molecular variance was performed on 675-base pair cytochrome oxidase I gene sequences from multiple populations of Hawaiian tree snails. Sequence data were analysed under distance based maximum-likelihood, and maximum-parsimony optimality criteria. Within the focal species, A. mustelina, numbers of variable and parsimony informative sites were 90 and 69, respectively. Pairwise intraspecific mtDNA sequence divergence ranged from 0 to 5.3% in A. mustelina, from 0 to 1.0% in A. livida and from 0 to 1.9% in A. sowerbyana. For A. mustelina, population genetic structure and mountain topography were strongly correlated. Maximum genetic distances were observed across deep, largely deforested valleys, and steep mountain peaks, independent of geographical distance. However, in certain areas where forest cover is presently fragmented, little mtDNA sequence divergence exists despite large geographical scales (8 km). Genetic data were used to define evolutionarily significant units for conservation purposes including decisions regarding placement of predator exclusion fences, captive propagation, re-introduction and translocation. PMID- 11918777 TI - A generalized heterozygote deficiency assessed with microsatellites in French common ash populations. AB - Common ash is a temperate forest tree with a colonizing behaviour, a discontinuous spatial distribution and a peculiar and poorly known mating system. Microsatellite markers were used to study the genetic structure in natural populations of common ash. Twelve populations located in northeastern France were analysed at five loci. Levels of genetic variability within and among stands were estimated for the seedling and adult stages. As expected for a forest tree, our results reveal high levels of intrapopulation diversity and a low genetic differentiation between stands. However, a general and significant heterozygote deficiency was found, with a mean F(IS) of 0.163 for the seedlings and of 0.292 for the adult trees. The different explanations for such an excess homozygosity are discussed: a nonMendelian inheritance of alleles, the presence of null alleles, a Wahlund effect and assortative mating. PMID- 11918778 TI - The discovery of three genetic races of the dwarf mistletoe Arceuthobium americanum (Viscaceae) provides insight into the evolution of parasitic angiosperms. AB - A population genetic approach was used to explore the evolutionary biology of the parasitic angiosperm Arceuthobium americanum Nutt. ex Engelm. (Viscaceae). Arceuthobium americanum infects three principal hosts and has the most extensive geographical range of any North American dwarf mistletoe. Based on the lack of apparent morphological and phenological differences between populations of A. americanum, past researchers have found no evidence for recognizing infraspecific taxa. In this study, molecular analysis using amplified fragment length polymorphism (AFLP) analysis indicated that A. americanum is divided into three distinct genetic races, each associated with a different host taxon in regions of allopatry: (i) Pinus banksiana in western Canada; (ii) Pinus contorta var. murrayana in the Sierra Nevada and Cascade Mountain ranges in the western US; and (iii) Pinus contorta var. latifolia in the western US and Canada. These observations suggest that host identity, geographical isolation and environmental factors have contributed to race formation in A. americanum. The lack of fine scale patterning within each of the A. americanum races is attributed to random dispersal of seeds over long distances by animal vectors. Historical factors such as glaciations and founder events have also influenced structuring and genetic diversity in A. americanum populations. Given sufficient time, it is possible that these races will become reproductively isolated and undergo speciation. PMID- 11918779 TI - The usefulness of amplified fragment length polymorphism markers for taxon discrimination across graduated fine evolutionary levels in Caribbean Anolis lizards. AB - Fine-level taxon discrimination is important in biodiversity assessment and ecogeographical research. Genomic markers are often required for studies on closely related taxa, however, most existing mitochondrial and nuclear markers require prior knowledge of the genome and are impractical for use in small conservation projects. This study describes the application of amplified fragment length polymorphism (AFLP) to discriminate at four progressively finer evolutionary levels of Caribbean Anolis lizards from the central Lesser Antilles. AFLP is shown to be a rapid and effective method for discriminating between species. Separation increases with primer pair number and choice of primer combination appears to be noncritical. Initial population-level results show markedly less discriminatory power. A screening technique for the identification of population informative markers combining principal component and principal coordinate analyses is presented and assessed. Subsequent results show selected conspecific AFLP data to be remarkably congruent with those of mitochondrial DNA, microsatellite and morphological markers. The use of AFLP as a low-cost nuclear marker in species-level taxon discrimination is supported, whereas population level application demands further consideration. PMID- 11918780 TI - Late Quaternary distributional stasis in the submediterranean mountain plant Anthyllis montana L. (Fabaceae) inferred from ITS sequences and amplified fragment length polymorphism markers. AB - Anthyllis montana is a submediterranean, herbaceous plant of the southern and central European mountains. The internal transcribed spacer (ITS) regions of nuclear ribosomal DNA were sequenced from multiple accessions of the species and several closely related taxa. In addition, amplified fragment length polymorphism (AFLP) was analysed from 71 individuals of A. montana collected in 20 localities, mainly in the Pyrenees, Alps, Italian Peninsula and Balkans. Our ITS phylogeny showed a sequential branching pattern in A. montana, implying a western Mediterranean origin followed by an eastward migration. ITS clock calibrations suggest that speciation of A. montana took place at the Pliocene-Pleistocene boundary, while intraspecific divergence dates to Late Quaternary times (i.e. 0.7 million years ago). The AFLP analyses revealed a major genetic (west/east) subdivision within A. montana, probably caused by the massive glaciation of the Alps during this latter time period. The present-day absence of A. montana from vast parts of the Alps, which appear ecologically suitable for the species, together with the finding of evenly distributed AFLP variability within each of the two western and eastern lineages identified, is taken as evidence for a largely static Late Quaternary history without large-scale migration. High levels of AFLP variation observed among populations, together with weak or absent patterns of isolation by distance, seem to be in accord with long-term population insularization and distributional stasis. However, recent small-scale migration and a narrow hybrid zone between western and eastern lineages need to be postulated to explain the intermediate genetic composition of individuals from the Maritime Alps, a well-known suture-zone for other plant and animal species. PMID- 11918781 TI - Population fragmentation in the southern rock agama, Agama atra: more evidence for vicariance in Southern Africa. AB - Mitochondrial DNA sequence data derived from two genes were used to infer phylogeographical relationships between 13 Agama atra populations. Three distinct geographical assemblages were found among the lizard populations. The first occurs in southern Namibia, the second is restricted to the western dry arid regions of South Africa, whereas the third is distributed throughout the more mesic southern and eastern parts of the subcontinent. Geographically structured differences among populations within Agama clades are probably the result of dispersal and historic isolations among populations. At the broader scale, there were marked congruences between the Agama genetic discontinuities and those described previously in other rock-dwelling vertebrates such as Pronolagus rupestris and Pachydactylus rugosus. This suggests vicariance, probably as a response to natural climatic changes during the past three million years. PMID- 11918782 TI - Amplified fragment length polymorphism analysis identifies hybrids between two subspecies of warblers. AB - In the western Pyrenees (Southwest France and Northwest Spain), a narrow hybrid zone exists between the common chiffchaff Phylloscopus (collybita) collybita and the Iberian chiffchaff Phylloscopus (c.) brehmii. In this zone, which is approximately 20 km wide, mixed matings and individuals singing the songs of both taxa occur at substantial frequencies (24 and 8.6%, respectively), suggesting frequent hybridization. Previous studies have shown very weak mitochondrial gene flow (Nm = 0.065), whereas four microsatellites suggested much higher nuclear gene flow (Nm = 4.9). In this study we used the amplified fragment length polymorphism (AFLP) method in order to identify hybrids and early backcrosses. We typed 91 birds from both allopatric and sympatric areas for 12 informative AFLP markers (of > 141 polymorphic fragments), obtained by screening 13 AFLP primer combinations. These individuals were previously typed for song (brehmii, collybita or mixed singers), mitochondrial DNA (mtDNA) haplotype and allelic genotypes at four microsatellite loci. Assignment tests demonstrated that in the zone of sympatry, a substantial number of intermediate genotypes existed among the birds previously believed to be pure collybita and brehmii, based on song and mtDNA haplotype. The majority of the mixed singers had intermediate genotypes. Our data suggest that the fraction of the adult population having a hybrid origin (hybrids or backcrosses) is in the order of 10%. With such a frequency of genetic hybrids, there would have been much more mtDNA introgression than observed, had female hybrids been perfectly fertile/viable. This result is consistent with male biased gene flow and Haldane's rule. PMID- 11918783 TI - Population genetic structure of Tomicus piniperda L. (Curculionidae: Scolytinae) on different pine species and validation of T. destruens (Woll.). AB - Genetic diversity and population structure of Tomicus piniperda was assessed using mitochondrial sequences on 16 populations sampled on 6 pine species in France. Amplifications of Internal transcribed space 1 (ITS1) were also performed. Our goals were to determine the taxonomic status of the Mediterranean ecotype T. piniperda destruens, and to test for host plant or geographical isolation effect on population genetic structure. We showed that T. piniperda clusters in two mtDNA haplotypic groups. Clade A corresponds to insects sampled in continental France on Pinus sylvestris, P. pinaster and P. uncinata, whereas clade B gathers the individuals sampled in Corsica on P. pinaster and P. radiata and in continental France on P. pinea and P. halepensis. Insects belonging to clade A and clade B also consistently differ in the length of ITS1. Individuals belonging to both clades were found once in sympatry on P. pinaster. Genetic distances between clades are similar to those measured between distinct species of Tomicus. We concluded that clade B actually corresponds to the destruens ecotype and forms a good species, T. destruens. Analyses of molecular variance ( amova ) were conducted separately on T. destruens and T. piniperda to test for an effect of either geographical isolation or host species. Interestingly, the effect of host plant was significant for T. piniperda only, while the effect of geographical isolation was not. Pine species therefore seems to act as a significant barrier to gene flow, even if host race formation is not observed. These results still need to be confirmed by nuclear markers. PMID- 11918784 TI - Genetic diversity in Arabidopsis thaliana L. Heynh. investigated by cleaved amplified polymorphic sequence (CAPS) and inter-simple sequence repeat (ISSR) markers. AB - In this study, we investigated genetic diversity among 37 accessions in Arabidopsis thaliana from Eurasia, North Africa and North America using morphological traits and two polymerase chain reaction (PCR)-based marker systems: cleaved amplified polymorphic sequences (CAPS) and inter-simple sequence repeats (ISSR). Cluster analysis based on genetic similarities calculated from CAPS data grouped the accessions roughly according to their geographical origin: one large group contained accessions from Western, Northern and Southern Europe as well as North Africa, a second group consisted of Eastern European and Asian continental accessions. North American accessions were interspersed into these groups. Contrary to the CAPS analysis, the dendrogram obtained from the ISSR data did not reflect the geographical origin of the accessions, and the calculated genetic distances did not match the CAPS results. This could be attributable to an uneven genomic distribution of ISSR markers as substantiated by a database search for ISSR binding sites in A. thaliana genomic DNA sequence files, or to the ISSR's different mode of evolution. We recommend CAPS markers for diversity analysis in A. thaliana because a careful selection of markers can ascertain an even representation of the entire genome. PMID- 11918785 TI - Social structure in migrating humpback whales (Megaptera novaeangliae). AB - Although largely solitary, humpback whales exhibit a number of behaviours where individuals co-operate with one another, for example during bubble net feeding. Such cases could be due to reciprocal altruism brought on by exceptional circumstances, for example the presence of abundant shoaling fish. An alternative explanation is that these behaviours have evolved through kin selection. With little restriction to either communication or movement, diffuse groups of relatives could maintain some form of social organization without the need to travel in tight-nit units. To try to distinguish between these hypotheses, we took advantage of the fact that migrating humpback whales often swim together in small groups. If kin selection is important in humpback whale biology, these groups should be enriched for relatives. Consequently, we analysed biopsy samples from 57 groups of humpback whales migrating off Eastern Australia in 1992. A total of 142 whales were screened for eight microsatellite markers. Mitochondrial DNA sequences (371 bp) were also used to verify and assist kinship identification. Our data add support to the notion that mothers travel with their offspring for the first year of the calf's life. However, beyond the presence of mother-calf/yearling pairs, no obvious relatedness pattern was found among whales sampled either in the same pod or on the same day. Levels of relatedness did not vary between migratory phases (towards or away from the breeding ground), nor between the two sexes considered either overall or in the north or south migrations separately. These findings suggest that, if any social organization does exist, it is formed transiently when needed rather than being a constant feature of the population, and hence is more likely based on reciprocal altruism than kin selection. PMID- 11918786 TI - Relatedness structure in Rhododendron metternichii var. hondoense revealed by microsatellite analysis. AB - The relatedness structure of Rhododendron metternichii Sieb. et Zucc. var. hondoense Nakai was analysed in a 150 x 70-m quadrat in Hiroshima Prefecture, western Japan. The population of R. metternichii occurred as three subpopulations at the study site (A1-A3). Pairwise relatedness based on microsatellite genotypes at eight loci and Mantel tests revealed a hierarchical structure of relatedness within and among subpopulations: (i) relatedness between individuals within 10 m of one another was significantly positive; (ii) relatedness between individuals in the same subpopulation was significantly positive, but negative between individuals in distant subpopulations; and (iii) relatedness was not significantly different from zero among neighbouring subpopulations. In detail, however, relatedness within each subpopulation was significantly positive in subpopulation A1, relatively weak but significantly positive in subpopulation A2, and not significantly different from zero in subpopulation A3. Relatedness within each subpopulation was inversely related to correlations between interindividual distance and relatedness. The aggregation of related individuals at short distances from one another may lead to decreasing relatedness within subpopulations as a whole. Moreover, negative correlations between interindividual distance and relatedness corresponded to high flowering densities at less than 10-m distance, implying that high flowering densities reduce pollinator foraging distance and lead to stronger genetic structure within subpopulations. Small individuals (< 2.0 m in height) showed stronger genetic structure compared with that of large individuals (> or = 2.0 m in height). The different relatedness structure within and among subpopulations may be caused by various degrees of gene flow affected by distribution patterns of individuals and population density. PMID- 11918787 TI - Genetic evidence for male biased dispersal in the red-billed quelea Quelea quelea. AB - In order to detect sex-biased dispersal in the red-billed quelea Quelea quelea in southern Africa, we used the assignment index technique to determine the probability that individuals originated from the population in which they were sampled. This is the first time that this multilocus genetic test has been used in a bird species and is informative despite evidence that the population under study exhibits little genetic structure. There was a pattern of male-biased dispersal, the first example in a passerine, and the first time that evidence of a sex-biased pattern of dispersal has been shown for queleas. PMID- 11918788 TI - Molecular determination of paternity in a natural population of the multiply mating polygynous lizard Eulamprus heatwolei. AB - We studied the mating system of the southern water skink, Eulamprus heatwolei, during spring and summer (encompassing the breeding season) in a population in southeastern Australia. We examined potential attributes that might influence the mating system and male reproductive success including home range size, physical proximity of adults and body size, and then genotyped all mothers, offspring and potential sires. Home range overlap of both sexes was extensive, with adult females sharing the greatest amount of space with each other and adult males the least amount of space with each other. However, not all adults hold home ranges. We classified approximately one quarter of adult males as home range holders and the rest as 'floaters'. Adult females occupy home ranges more than males, with approximately three-quarters classified as home range holders. Home range ownership is not correlated with body size for either sex, however, male body size is positively correlated with the number of adult female home ranges that his home range overlaps and adult male home ranges are larger than those of females. We used microsatellite genotyping to assign paternities to 55 offspring from 17 litters and then compared this data with our home range and behavioural observations. This species displays extreme levels of multiple paternity given the small mean clutch size of three. Multiple paternity was confirmed in 11 (64.7%) of 17 clutches but three other clutches (for a total of 82.4%) also may display multiple paternity. A total of 30 offspring from 12 litters were assigned to 10 of the 32 genotyped adult males from our study site. Of these 10 adult males, half were home range holders. Five complete clutches and a total of 25 out of the 55 offspring could not be positively assigned to any male surveyed as part of the study and were attributed to floater males or resident males adjacent to our study site that had not been genotyped. While sample sizes are small, neither male home range ownership nor body size is significantly correlated with the number of paternities a male obtained. Our study suggests a polygynous mating system for this species. PMID- 11918789 TI - Vertical transmission of sublethal granulovirus infection in the Indian meal moth, Plodia interpunctella. AB - Knowledge of the mechanisms of pathogen persistence in relation to fluctuations in host density is crucial to our understanding of disease dynamics. In the case of insect baculoviruses, which are typically transmitted horizontally via a lifestage that can persist outside the host, a key issue that remains to be elucidated is whether the virus can also be transmitted vertically as a sublethal infection. We show that RNA transcripts for the Plodia interpunctella GV granulin gene are present in a high proportion of P. interpunctella insects that survive virus challenge. Granulin is a late-expressed gene that is only transcribed after viral genome replication, its presence thus strongly indicates that viral genome replication has occurred. Almost all insects surviving the virus challenge tested positive for viral RNA in the larval and pupal stage. However, this proportion declined in the emerging adults. Granulin mRNA was also detected in both the ovaries and testes, which may represent a putative mechanism by which reduced fecundity in sublethally affected hosts might be manifested. RNA transcripts were also detected in 60-80% of second-generation larvae that were derived from mating surviving adults, but there was no difference between the sexes, with both males and females capable of transmitting a sublethal infection to their offspring. The data indicate that low-level persistent infection, with at least limited gene expression, can occur in P. interpunctella following survival of a granulovirus challenge. We believe that this is the first demonstration of a persistent, sublethal infection by a baculovirus to be initiated by a sublethal virus dose. We hypothesize that the 'latent' baculovirus infections frequently referred to in the literature may also be low level persistent, sublethal infections resulting from survival from initial baculovirus exposure. PMID- 11918790 TI - Fine-level mycorrhizal specificity in the Monotropoideae (Ericaceae): specificity for fungal species groups. AB - The Monotropoideae (Ericaceae) are non-photosynthetic angiosperms that obtain fixed carbon from basidiomycete ectomycorrhizal fungi. In previous work, we showed that each plant species is associated with a single genus or a set of closely related genera of ectomycorrhizal fungi. Here we show that the level of specificity is much higher. We used a molecular phylogenetic approach to contrast specificity patterns among eight plant lineages and three fungal genera. We relied on fungal nuclear internal transcribed spacer (nrITS) sequence data obtained from 161 basidiocarps and 85 monotropoid roots representing 286 sampled plants screened using restriction length polymorphisms. From the phylogenetic placement of fungal symbionts in fungal phylograms, we found that three basal (Sarcodes, Pterospora, Pleuricospora) and one derived lineage (Allotropa) of plants target narrow clades of closely related species groups of fungi, and four derived lineages (Monotropa hypopithys species group, Pityopus) target more distant species groups. Within most plant lineages, geography and photobiont association constrain specificity. Specificity extended further in Pterospora andromedea, in which sequence haplotypes at the plastid trn L-F region of 73 plants were significantly associated with different fungal species groups even in sympatry. These results indicate that both the macro- and microevolution of the Monotropoideae are tightly coupled to their mycorrhizal symbionts. PMID- 11918791 TI - Testing for recombination in a short nuclear DNA sequence of the European meadow grasshopper, Chorthippus parallelus. AB - Single-copy nuclear DNA sequences have high potential as a source of genetic markers for population analyses. However, the difficulties that arise when haplotypes that are the product of recombinational rearrangements are present require additional consideration. Two statistical methods for identifying potential recombinants by detecting anomalies in the distribution of variable sites along sequences were used to screen sequences from a single-copy nuclear DNA fragment, cpnl-1, of the European meadow grasshopper (Chorthippus parallelus). Five of the 71 haplotypes in the cpnl-1 data set showed nonrandom distribution of polymorphic sites using both methods. The second method pinpointed an additional four haplotypes. Estimates of the rate of recombination in the entire data set were obtained using standard methods. It is concluded that cpnl-1 haplotypes have been involved in recombination or gene conversion events at a rate more than twice the mutation rate. This confirms that recombination and gene conversion are significant factors in the generation of haplotype variation in nuclear gene sequences. The cpnl-1 haplotypes identified by the tests were present only in populations that have had recent contact; the Balkan and Turkish refugial populations and their post-glacial colonies to the north. This is discussed in relation to the phylogenetic inferences drawn from the same data in a previous report. PMID- 11918792 TI - Cross-species microsatellite markers for elucidating population genetic structure in Arabidopsis and Arabis (Brassicaeae). AB - Species closely related to model organisms present the opportunity to efficiently apply molecular and functional tools developed by a large research community to taxa with different ecological and evolutionary histories. We complied 42 microsatellite loci that amplify under common conditions in four closely related Arabidopsis: A. thaliana; A. halleri; A. lyrata ssp. lyrata; and A. lyrata ssp. petraea, as well as in one more distantly related crucifer; Arabis drummondii. Variation at these loci is amenable to a diversity of applications including population genetics, phylogeographical analyses, mapping of inter and intraspecific crosses, and recombination mapping. Our analysis of microsatellite variation illustrates significant differences in population genetic parameters among three Arabidopsis species. A population of A. thaliana, an inbreeding annual plant associated with disturbed habitats, was highly monomorphic (P = 8% percent polymorphic loci) and only 0.2% heterozygous for 648 locus-by-individual combinations. A population of the self-incompatible perennial herb, A. halleri, was more genetically variable (P = 71%) and had an excess of heterozygosity that may reflect a recent population bottleneck associated with human-mediated founder events. A population of the self-incompatible perennial herb, A. lyrata ssp. petraea, was even more genetically variable (P = 86%) and appeared to be at mutation-drift equilibrium. Population structure estimated from neutrally evolving loci provides an empirical expectation against which hypotheses of adaptive evolution at functional loci can be tested. PMID- 11918794 TI - Maternal inheritance of a chloroplast microsatellite marker in controlled hybrids between Fraxinus excelsior and Fraxinus angustifolia. AB - Restriction fragment length polymorphism, polymerase chain reaction-restriction fragment length polymorphism and simple sequence repeat (SSR or microsatellites) analyses were performed to detect chloroplast DNA polymorphisms between two ash species, Fraxinus excelsior and F. angustifolia. Only one SSR locus was found to be polymorphic, confirming the very close relatedness of these species. Inheritance of this marker was studied in hybrids obtained from controlled crosses between the two tree species. Results indicated, for the first time in Oleaceae, that chloroplasts are maternally inherited. This chloroplast SSR marker is now used concomitantly with nuclear markers to analyse ash populations in sympatric areas. PMID- 11918793 TI - Single-nucleotide polymorphism characterization in species with limited available sequence information: high nucleotide diversity revealed in the avian genome. AB - As a case study for single-nucleotide polymorphism (SNP) identification in species for which little or no sequence information is available, we investigated several approaches to identifying SNPs in two passerine bird species: pied and collared flycatchers (Ficedula hypoleuca and F. albicollis). All approaches were successful in identifying sequence polymorphism and over 50 candidate SNPs per species were identified from approximately 9.1 kb of sequence. In addition, 17 sites were identified in which the frequency of alternative bases differed by > 50% between species (termed interspecific SNPs). Interestingly, polymorphism of microsatellite/intron loci in the source species appeared to be a positive predictor of nucleotide diversity in homologous flycatcher sequences. The overall nucleotide diversity of flycatchers was 2.3-2.7 x 10(-3), which is approximately 3-6 times higher than observed in recent studies of human SNPs. Higher nucleotide diversity in the avian genome could be due to the relatively older age of flycatcher populations, compared with humans, and/or a higher long-term effective population size. PMID- 11918795 TI - Speciation via introgressive hybridization in East African cichlids? AB - Speciation caused by introgressive hybridization occurs frequently in plants but its importance remains controversial in animal evolution. Here we report a case of introgressive hybridization between two ancient and genetically distinct species of Lake Tanganyika cichlids that led to the formation of a new species. Neolamprologus marunguensis contains mtDNA haplotypes from both parental species varying on average by 12.4% in the first section of the control region and by 5.2% in a segment of the cytochrome b gene. All individuals have almost identical DNA sequences in the flanking regions of the single-copy nuclear DNA locus TmoM27, and show a mosaic of alleles derived from both parental lineages in six microsatellite loci. Hence, our finding displays another mode of speciation in cichlid fishes. The increase of genetic and phenotypic diversity due to hybridization may contribute to the uniquely rapid pace of speciation in cichlids. PMID- 11918796 TI - Osmotic stress-coupled maintenance of polar growth in Aspergillus nidulans. AB - Free-living cells monitor extracellular 'osmotic strength' and respond metabolically to offset unfavourable osmotic intracellular solute concentrations. Here, we report the reconstruction of the Aspergillus nidulans salt stress controlling MAP kinase pathway, based on homology analysis with known yeast genes. In A. nidulans, salt stress HOG genes, such as pbsA, hogA, ptpA and msnA, are upregulated when exposed to high concentrations of salt and, in a hogA deletion mutant (SIK1), the accumulation of pbsA is strongly reduced, suggesting a salt-specific feedback induction mechanism. Growth of SIK1 appears to be unchanged in unstressed cells, but hyphal extension rates are reduced by as much as 60% in the presence of salt. Microscopic observation revealed abnormal hyperbranched hyphal tips, disproportionate accumulation of nuclei and absence of septa. Thus, the inability to maintain turgor pressure depresses cell expansion and results in slower volume increases. In addition, SIK1 fails to partition the apical cell; thus, nuclei are not likely to arrest mitosis in interphase as in normal cells, but continue to divide, accumulating to high levels. PMID- 11918797 TI - The MotA transcription factor from bacteriophage T4 contains a novel DNA-binding domain: the 'double wing' motif. AB - MotA is a transcription factor from bacteriophage T4 that helps adapt the host Escherichia coli transcription apparatus to T4 middle promoters. We have determined the crystal structure of the C-terminal DNA-binding domain of MotA (MotCF) to 1.6 A resolution using multiwavelength, anomalous diffraction methods. The structure reveals a novel DNA-binding alpha/beta motif that contains an exposed beta-sheet surface that mediates interactions with the DNA. Independent biochemical experiments have shown that MotCF binds to one surface of a single turn of DNA through interactions in adjacent major and minor grooves. We present a model of the interaction in which beta-ribbons at opposite corners of the six stranded beta-sheet penetrate the DNA grooves, and call the motif a 'double wing' to emphasize similarities to the 'winged-helix' motif. The model is consistent with data on how MotA functions at middle promoters, and provides an explanation for why MotA can form non-specific multimers on DNA. PMID- 11918798 TI - Salmonella effectors within a single pathogenicity island are differentially expressed and translocated by separate type III secretion systems. AB - Pathogenicity islands (PAIs) are large DNA segments in the genomes of bacterial pathogens that encode virulence factors. Five PAIs have been identified in the Gram-negative bacterium Salmonella enterica. Two of these PAIs, Salmonella pathogenicity island (SPI)-1 and SPI-2, encode type III secretion systems (TTSS), which are essential virulence determinants. These 'molecular syringes' inject effectors directly into the host cell, whereupon they manipulate host cell functions. These effectors are either encoded with their respective TTSS or scattered elsewhere on the Salmonella chromosome. Importantly, SPI-1 and SPI-2 are expressed under distinct environmental conditions: SPI-1 is induced upon initial contact with the host cell, whereas SPI-2 is induced intracellularly. Here, we demonstrate that a single PAI, in this case SPI-5, can encode effectors that are induced by distinct regulatory cues and targeted to different TTSS. SPI 5 encodes the SPI-1 TTSS translocated effector, SigD/SopB. In contrast, we report that the adjacently encoded effector PipB is part of the SPI-2 regulon. PipB is translocated by the SPI-2 TTSS to the Salmonella-containing vacuole and Salmonella-induced filaments. We also show that regions of SPI-5 are not conserved in all Salmonella spp. Although sigD/sopB is present in all Salmonella spp., pipB is not found in Salmonella bongori, which also lacks a functional SPI 2 TTSS. Thus, we demonstrate a functional and regulatory cross-talk between three chromosomal PAIs, SPI-1, SPI-2 and SPI-5, which has significant implications for the evolution and role of PAIs in bacterial pathogenesis. PMID- 11918799 TI - Rpa12p, a conserved RNA polymerase I subunit with two functional domains. AB - Rpa12p is a subunit of RNA polymerase I formed of two zinc-binding domains. The N terminal zinc region (positions 1-60) is poorly conserved from yeast to man. The C-terminal domain contains an invariant Q.RSADE.T.F motif shared with the TFIIS elongation factor of RNA polymerase II and its archaeal counterpart. Deletions removing the N-terminal domain fail to grow at 34 degrees C, are sensitive to nucleotide-depleting drugs and become lethal in rpa14-Delta mutants lacking the non-essential RNA polymerase I subunit Rpa14p. They also strongly alter the immunofluorescent properties of RNA polymerase I in the nucleolus. Finally, they prevent the binding of Rpa12p to immunopurified polymerase I and impair a specific two-hybrid interaction with the second largest subunit. In all these respects, N-terminal deletions behave like full deletions. In contrast, C terminal deletions retaining only the first N-terminal 60 amino acids are indistinguishable from wild type. Thus, the N-terminal zinc domain of Rpa12p determines its anchoring to RNA polymerase I and is the only critical part of that subunit in vivo. PMID- 11918800 TI - The Sinorhizobium meliloti stringent response affects multiple aspects of symbiosis. AB - Sinorhizobium meliloti and host legumes enter into a nitrogen-fixing, symbiotic relationship triggered by an exchange of signals between bacteria and plant. S. meliloti produces Nod factor, which elicits the formation of nodules on plant roots, and succinoglycan, an exopolysaccharide that allows for bacterial invasion and colonization of the host. The biosynthesis of these molecules is well defined, but the specific regulation of these compounds is not completely understood. Bacteria control complex regulatory networks by the production of ppGpp, the effector molecule of the stringent response, which induces physiological change in response to adverse growth conditions and can also control bacterial development and virulence. Through detailed analysis of an S. meliloti mutant incapable of producing ppGpp, we show that the stringent response is required for nodule formation and regulates the production of succinoglycan. Although it remains unknown whether these phenotypes are connected, we have isolated suppressor strains that restore both defects and potentially identify key downstream regulatory genes. These results indicate that the S. meliloti stringent response has roles in both succinoglycan production and nodule formation and, more importantly, that control of bacterial physiology in response to the plant and surrounding environment is critical to the establishment of a successful symbiosis. PMID- 11918801 TI - Overexpression of the putative thiol conjugate transporter TbMRPA causes melarsoprol resistance in Trypanosoma brucei. AB - Melaminophenyl arsenical drugs are a mainstay of chemotherapy against late-stage African sleeping sickness, but drug resistance is increasingly prevalent. We describe here the characterization of two genes encoding putative metal-thiol conjugate transporters from Trypanosoma brucei. The two proteins, TbMRPA and TbMRPE, were each overexpressed in trypanosomes, with or without co-expression of two key enzymes in trypanothione biosynthesis, ornithine decarboxylase and gamma glutamyl-cysteine synthetase. Overexpression of gamma-glutamyl-cysteine synthetase resulted in a twofold increase in cellular trypanothione, whereas overexpression of ornithine decarboxylase had no effect on the trypanothione level. The overexpression of TbMRPA resulted in a 10-fold increase in the IC50 of melarsoprol. The overexpression of the trypanothione biosynthetic enzymes alone gave two- to fourfold melarsoprol resistance, but did not enhance resistance caused by MRPA. Overexpression of TbMRPE had little effect on susceptibility to melarsoprol but did give two- to threefold resistance to suramin. PMID- 11918802 TI - The C-terminus of IcmT is essential for pore formation and for intracellular trafficking of Legionella pneumophila within Acanthamoeba polyphaga. AB - We have shown previously that the five rib (release of intracellular bacteria) mutants of Legionella pneumophila are competent for intracellular replication but defective in pore formation-mediated cytolysis and egress from protozoan and mammalian cells. The rib phenotype results from a point mutation (deletion) DeltaG544 in icmT that is predicted to result in the expression of a protein truncated by 32 amino acids from the C-terminus. In contrast to the rib mutants that are capable of intracellular replication, an icmT null mutant was completely defective in intracellular replication within mammalian and protozoan cells, in addition to its defect in pore formation-mediated cytolysis. The icmT wild-type allele complemented the icmT null mutant for both defects of intracellular replication and pore formation-mediated cytolysis and egress from mammalian cells. In contrast, the icmTDeltaG544 allele complemented the icmT null mutant for intracellular growth, but not for the pore-forming activity. Consistent with their defect in pore formation-mediated cytotoxicity in vitro, both mutants failed to cause pulmonary inflammation in A/J mice. Interestingly, the rib mutant was severely defective in intracellular growth within Acanthamoeba polyphaga. Confocal laser scanning and electron microscopy confirmed that the rib mutant and the icmT null mutant were severely and completely defective, respectively, in intracellular growth in A. polyphaga, and the respective defects correlated with fusion of the bacterial phagosomes to lysosomes. Taken together, the data showed that the C-terminus domain of IcmT is essential for the pore-forming activity and is required for intracellular trafficking and replication within A. polyphaga, but not within mammalian cells. PMID- 11918803 TI - Antigenic variation of Anaplasma marginale msp2 occurs by combinatorial gene conversion. AB - The rickettsial pathogen Anaplasma marginale establishes lifelong persistent infection in the mammalian reservoir host, during which time immune escape variants continually arise in part because of variation in the expressed copy of the immunodominant outer membrane protein MSP2. A key question is how the small 1.2 Mb A. marginale genome generates sufficient variants to allow long-term persistence in an immunocompetent reservoir host. The recombination of whole pseudogenes into the single msp2 expression site has been previously identified as one method of generating variants, but is inadequate to generate the number of variants required for persistent infection. In the present study, we demonstrate that recombination of a whole pseudogene is followed by a second level of variation in which small segments of pseudogenes recombine into the expression site by gene conversion. Evidence for four short sequential changes in the hypervariable region of msp2 coupled with the identification of nine pseudogenes from a single strain of A. marginale provides for a combinatorial number of possible expressed MSP2 variants sufficient for lifelong persistence. PMID- 11918804 TI - Identification of binding sites for the group A streptococcal global regulator CovR. AB - The CovRS two-component system (also called CsrRS) of the group A streptococcus (GAS) acts as a global regulator, influencing the transcription of at least six virulence factors. The synthesis of the hyaluronic acid capsule, a virulence factor encoded by the hasABC operon, is negatively regulated by CovRS. We confirmed that phosphorylation of CovR increases its binding to a DNA fragment containing the hasA promoter. Using DNase I footprinting, we identified five binding sites surrounding the hasA promoter from bases -79 to +73 (where +1 is the start of transcription). One pair of thymines within each binding site appears to be necessary for CovR binding in vitro, as shown by uracil interference analysis. When each of these thymine pairs was altered by site directed mutagenesis, CovR binding was reduced in vitro, confirming the role of each thymine pair in binding. Using a transcriptional reporter system with a single chromosomal copy of PhasA-gusA, we demonstrated the importance of each of four of these binding sites for CovR repression of the hasA promoter. Based on this information, we propose a consensus sequence for CovR binding to DNA. PMID- 11918805 TI - The WD40-repeat protein CreC interacts with and stabilizes the deubiquitinating enzyme CreB in vivo in Aspergillus nidulans. AB - Genetic dissection of carbon catabolite repression in Aspergillus nidulans has identified two genes, creB and creC, which, when mutated, affect expression of many genes in both carbon catabolite repressing and derepressing conditions. The creB gene encodes a functional deubiquitinating enzyme and the creC gene encodes a protein that contains five WD40 repeat motifs, and a proline-rich region. These findings have allowed the in vivo molecular analysis of a cellular switch involving deubiquitination. We demonstrate that overexpression of the CreB deubiquitinating enzyme can partially compensate for a lack of the CreC WD40 repeat protein in the cell, but not vice versa and, thus, the CreB deubiquitinating enzyme acts downstream of the CreC WD40-repeat protein. We demonstrate using co-immunoprecipitation experiments that the CreB deubiquitinating enzyme and the CreC WD40-repeat protein interact in vivo in both carbon catabolite repressing and carbon catabolite derepressing conditions. Further, we show that the CreC WD40-repeat protein is required to prevent the proteolysis of the CreB deubiquitinating enzyme in the absence of carbon catabolite repression. This is the first case in which a regulatory deubiquitinating enzyme has been shown to interact with another protein that is required for the stability of the deubiquitinating enzyme. PMID- 11918806 TI - Control of chitin synthesis through Shc1p, a functional homologue of Chs4p specifically induced during sporulation. AB - The Saccharomyces cerevisiae SHC1 gene encodes a protein with a high homology to Chs4p, a positive regulator of chitin synthase III (CSIII) during vegetative growth. SHC1 is not expressed during vegetative growth but is strongly induced during sporulation as a mid-late gene. shc1/shc1 mutants do not show any defect in the total rate of sporulation and meiosis occurs normally. However, shc1/shc1 ascospores be-come highly permeable to DAPI, much more sensitive to glusulase treatment, and have very low levels of chitosan in their cell walls. All these observations indicate that Shc1p is required for proper maturation of the ascospore through its participation in the synthesis of the chitosan layer. Lack of SHC1 during sporulation can be partially compensated by over-expression of the CHS4 gene. During vegetative growth, SHC1 has no apparent function but, when ectopically overexpressed, it can substitute Chs4p as an activator of the CSIII activity; however, Shc1p fails to localize it properly, as Chs4p does. In conclusion, S. cerevisiae contains two functionally redundant genes in the control of CSIII activity: CHS4, whose function is restricted to vegetative growth because Chs4p is rapidly degraded during sporulation, and SHC1, whose function in cell wall ascospore assembly is transcriptionally restricted to the sporulation process. PMID- 11918807 TI - A common drug-responsive element mediates the upregulation of the Candida albicans ABC transporters CDR1 and CDR2, two genes involved in antifungal drug resistance. AB - Upregulation of the ATP-binding cassette (ABC) transporter genes CDR1 and CDR2 (Candida drug resistance 1 and 2) is a common mechanism observed in Candida albicans clinical isolates developing resistance to the class of azole antifungals. In this work, the regulatory elements of both genes were delimited using a reporter system in an azole-susceptible strain exposed to oestradiol, which allows transient induction of these genes. We found two regulatory elements in the CDR1 promoter: one responsible for basal expression (basal expression element; BEE) and the other required for oestradiol responsiveness (drug responsive element I; DREI). In the CDR2 promoter, a single regulatory element responsible for oestradiol responsiveness (DREII) was detected. Both DREs shared a consensus of 21 bp with the sequence 5'-CGGA(A/T)ATCGGATATTTTTTTT-3' having no equivalent to known eukaryotic regulatory sequence. Consistent with this finding, two other C. albicans genes identified by a search for the presence of DRE in the C. albicans genome sequence database were responsive to oestradiol. Finally, the regulatory elements found in CDR1 and CDR2 were also functional in an azole resistant strain with constitutive high expression of both transporters. These results suggest that, although CDR1 and CDR2 upregulation can be obtained by transient drug-induced and constitutive upregulation, these two processes converge to the same regulatory elements and probably mobilize the same trans acting factors. PMID- 11918808 TI - Engineering specificity of starter unit selection by the erythromycin-producing polyketide synthase. AB - Chain initiation on many modular polyketide synthases is mediated by acyl transfer from the CoA ester of a dicarboxylic acid, followed by decarboxylation in situ by KSQ, a ketosynthase-like decarboxylase domain. Consistent with this, the acyltransferase (AT) domains of all KSQ-containing loading modules are shown here to contain a key arginine residue at their active site. Site-specific replacement of this arginine residue in the oleandomycin (ole) loading AT domain effectively abolished AT activity, consistent with its importance for catalysis. Substitution of the ole PKS loading module, or of the tylosin PKS loading module, for the erythromycin (ery) loading module gave polyketide products almost wholly either acetate derived or propionate derived, respectively, instead of the mixture found normally. An authentic extension module AT domain, rap AT2 from the rapamycin PKS, functioned appropriately when engineered in the place of the ole loading AT domain, and gave rise to substantial amounts of C13 methylerythromycins, as predicted. The role of direct acylation of the ketosynthase domain of ex-tension module 1 in chain initiation was confirmed by demonstrating that a mutant of the triketide synthase DEBS1-TE, in which the 4' phosphopante-theine attachment site for starter acyl groups was specifically removed, produced triketide lactone pro-ducts in detectable amounts. PMID- 11918809 TI - A tyrosine-phosphorylated 12-amino-acid sequence of enteropathogenic Escherichia coli Tir binds the host adaptor protein Nck and is required for Nck localization to actin pedestals. AB - Enteropathogenic Escherichia coli (EPEC) and enterohaemorrhagic E. coli (EHEC) each promote the reorganization of actin into filamentous pedestal structures beneath attached bacteria during colonization of the intestinal epithelium. Central to this process is the translocation of the protein Tir (translocated intimin receptor) into the plasma membrane of host cells, where it interacts with the bacterial outer membrane protein intimin and triggers cellular signalling events that lead to actin rearrangement. Actin signalling by EPEC Tir requires a tyrosine residue, Y474, which is phosphorylated in the host cell. In contrast, EHEC Tir lacks this residue and generates pedestals independently of tyrosine phosphorylation. Consistent with this difference, recent work indicates that EHEC Tir cannot functionally replace EPEC Tir. To identify the role that tyrosine phosphorylation of EPEC Tir plays in actin signalling, we generated chimeric EHEC/EPEC Tir proteins and identified a 12-residue sequence of EPEC Tir containing Y474 that confers actin-signalling capabilities to EHEC Tir when the chimera is expressed in EPEC. Nck, a mammalian adaptor protein that has been implicated in the initiation of actin signalling, binds to this sequence in a Y474 phosphorylation-dependent manner and is recruited to the pedestals of EPEC, but not of EHEC. PMID- 11918810 TI - Functional complementation reveals the importance of intermolecular monomer interactions for Helicobacter pylori VacA vacuolating activity. AB - The Helicobacter pylori vacuolating cytotoxin (VacA) induces degenerative vacuolation of sensitive mammalian cell lines. Although evidence is accumulating that VacA enters cells and functions from an intracellular site of action, the biochemical mechanism by which VacA mediates cellular vacuolation has not been established. In this study, we used functional complementation and biochemical approaches to probe the structure of VacA. VacA consists of two discrete fragments, p37 and p58, that are both required for vacuolating activity. Using a transient transfection system, we expressed genetically modified forms of VacA and identified mutations in either p37 or p58 that inactivated the toxin. VacA with an inactivating single-residue substitution in the p37 domain [VacA (P9A)] functionally complemented a second mutant form of VacA with an inactivating two residue deletion in the p58 domain [VacA Delta(346-347)]. VacA (P9A) and VacA Delta(346-347) also co-immunoprecipitated from vacuolated monolayers, supporting the hypothesis that these two inactive mutants associate directly to function in trans. p37 and p58 interact directly when expressed as separate fragments within HeLa cells, suggesting that p37-p58 inter-actions facilitate VacA monomer associations. Collectively, these results support a model in which the active form of VacA requires assembly into a complex of two or more monomers to elaborate toxin function. PMID- 11918811 TI - The fle1 gene encoding a C2H2 zinc finger protein co-ordinates male and female sexual differentiation in Podospora anserina. AB - The flexuosa (fle1-1) mutant, isolated in Podospora anserina, displays vegetative defects and two antagonistic sexual phenotypes: it produces several 1000-fold fewer microconidia (male gametes) than the wild-type strain and, conversely, more abundant protoperithecia (female organs). Cloning and sequencing of the fle1 gene and of cDNA identified an open reading frame encoding a 382-amino-acid polypeptide with two C2H2 zinc finger motifs. The predicted FLE1 protein shares 46% identity with the FlbC protein of Aspergillus nidulans and 68% identity with a putative protein identified by a search in the Neurospora crassa database. The nuclear localization of FLE1 was demonstrated by fusion with the green fluorescent protein. Sequencing of the fle1-1 mutant allele revealed a frameshift mutation upstream of the zinc finger domain. The fle1-1 mutant was a null mutant, as targeted disruption of fle1 sequence led to the same pleiotropic phenotype. When fle1 was overexpressed by introduction of a transgenic copy of the native fle1 gene or a fusion with a strong promoter, formation of protoperithecia was impaired, leading to partial or complete female sterility. We propose that fle1 acts as a repressor of female sexual differentiation in order to maintain the balance between male and female sexual pathways. PMID- 11918812 TI - Examination of Salmonella gene expression in an infected mammalian host using the green fluorescent protein and two-colour flow cytometry. AB - Quantitative data on Salmonella gene expression in infected hosts are largely lacking because of technical problems. One attractive reporter, the green fluorescent protein (GFP), is widely used in vitro but is difficult to quantify in infected tissues because of the preponderance of background particles with similar fluorescence. Here, bacterial GFP emission was spectrally distinguished from host autofluorescence by two-colour flow cytometry. Using this technique, the in vivo activity of three well-characterized promoters (PsicA, PssaH and PpagC) was determined. Their spatial and temporal activity patterns are in close agreement with predictions based on previous data and the colonization defects of corresponding deletion strains. To identify additional Salmonella promoters that are induced in infected animals, a genomic library was sorted by flow cytometry yielding four independent promoters. Genes expressed from PpibB and PsifA contribute to virulence, and chorismate mutase expressed from ParoQ might participate in aromatic acid biosynthesis, which is also required for virulence. Promoter P3g appears to be part of a mobile genetic element that is lacking in the completely sequenced strain LT2. PMID- 11918813 TI - Transcription of multiple var genes by individual, trophozoite-stage Plasmodium falciparum cells expressing a chondroitin sulphate A binding phenotype. AB - In this study, we detected multiple var gene transcripts within single, mature trophozoite-infected red blood cells (iRBCs) bound to chondroitin sulphate A (CSA). Several of the var detected had previously been demonstrated to encode Plasmodium falciparum erythrocyte membrane protein-1 (PfEMP-1) variants with domains that mediated iRBC adhesion to receptors other than CSA. Parasites expressing the CSA-adherent phenotype transcribed far more of one var than of all others, but this gene was different from the two other var previously purported to encode adhesion to CSA. Previous work suggesting that only single var are transcribed by mature trophozoites needs re-examination in the light of these data from single, infected cells. PMID- 11918815 TI - An rRNA mutation identifies the apicoplast as the target for clindamycin in Toxoplasma gondii. AB - Toxoplasma gondii is a protozoan sensitive to several inhibitors of prokaryotic translation (e.g. clindamycin, macrolides and tetracyclines). A priori, two prokaryotic-like organelles, the 'apicoplast' (a non-photosynthetic plastid) and the mitochondrion, are likely targets for these drugs. Without using overt mutagenesis, we selected two independent clones (ClnR-4 and ClnR-21) with strong and stable clindamycin resistance. Several lines with substantial but lower levels of resistance were also isolated with (XR-46) or without (ClnR-23) overt mutagenesis. The ClnR-4 and ClnR-21 mutants uniquely possess a G-->U point mutation at position 1857 of the apicoplast large-subunit rRNA, whereas no mutation was identified in this region for ClnR-23 or XR-46. Position 1857 corresponds to position 2061 in Escherichia coli where it is predicted to bind clindamycin. The mutation is present in all the apicoplast rDNA copies (an estimated 12 per organelle), indicative of a strong selective advantage in the presence of clindamycin. In the absence of drug, however, such a mutation is unlikely to be neutral, as the G is a critical contributor to the transpeptidation reaction and absolutely conserved in all kingdoms. This may explain why ClnR-4 shows a slight growth defect in vitro. These mutants provide direct genetic evidence that apicoplast translation is the target for clindamycin in Toxoplasma. Further, their sensitivity profiles to other antibiotics specific for the large ribosomal subunit (macrolides and chloramphenicol) and, intriguingly, the small subunit (doxycycline) argue that these drugs also target the apicoplast ribosome. PMID- 11918814 TI - Control of 26S proteasome expression by transcription factors regulating multidrug resistance in Saccharomyces cerevisiae. AB - In eukaryotic cells, intracellular proteolysis occurs mainly via the ubiquitin proteasome system. Expression of the yeast proteasome is under the control of the transcription factor, Rpn4p (also known as Son1p/Ufd5p). We show here that the RPN4 gene promoter contains regulatory sequences that bind Pdr1p and Pdr3p, two homologous zinc finger-containing transcription factors, which mediate multiple drug resistance through the expression of membrane transporter proteins. Mutations in the RPN4 Pdr1p/Pdr3p binding sites lead to decreased expression of the proteasome RPT6 gene and to defective ubiquitin-mediated proteolysis. Pdr3p, but not Pdr1p, is required for normal levels of intracellular proteolysis, indicating that the two transcription factors have distinct functions in the control of RPN4 expression. The RPN4 promoter contains an additional sequence that binds Yap1p, a bZIP-type transcription factor that plays an important role in the oxidative stress response and multidrug resistance. We also show that the Yap1p response element is important in the transactivation of RPN4 by Yap1p. In yeast cells lacking Pdr1p, ubiquitin-Pro-beta-galactosidase, a short-lived protein used to assay proteasome activity, is stabilized by the loss of Yap1p. These data demonstrate that the ubiquitin-proteasome system is controlled by transcriptional regulators of multidrug resistance via RPN4 expression. PMID- 11918816 TI - Endonuclease cleavage of messenger RNA in Bacillus subtilis. AB - A deletion derivative of the ermC gene was constructed that expresses a 254 nucleotide mRNA. The small size of this mRNA facilitated the detection of processing products that did not differ greatly in size from the full-length transcript. In the presence of erythromycin, which induces ribosome stalling near the 5' end of ermC mRNA, the 254-nucleotide mRNA was cleaved endonucleolytically at the site of ribosome stalling. Only the downstream product of this cleavage was detectable; the upstream product was apparently too unstable to be detected. The downstream cleavage product accumulated at times after rifampicin addition, suggesting that the stalled ribosome at the 5' end conferred stability to this RNA fragment. Neither Bs-RNase III nor RNase M5, the two known narrow-specificity endoribonucleases of Bacillus subtilis, was responsible for this cleavage. These results indicate the presence in B. subtilis of another specific endoribonuclease, which may be ribosome associated. PMID- 11918817 TI - Microarray analysis of the Bacillus subtilis K-state: genome-wide expression changes dependent on ComK. AB - In Bacillus subtilis, the competence transcription factor ComK activates its own transcription as well as the transcription of genes that encode DNA transport proteins. ComK is expressed in about 10% of the cells in a culture grown to competence. Using DNA microarrays representing approximately 95% of the protein coding open reading frames in B. subtilis, we compared the expression profiles of wild-type and comK strains, as well as of a mecA mutant (which produces active ComK in all the cells of the population) and a comK mecA double mutant. In these comparisons, we identified at least 165 genes that are upregulated by ComK and relatively few that are downregulated. The use of reporter fusions has confirmed these results for several genes. Many of the ComK-regulated genes are organized in clusters or operons, and 23 of these clusters are preceded by apparent ComK box promoter motifs. In addition to those required for DNA uptake, other genes that are upregulated in the presence of ComK are probably involved in DNA repair and in the uptake and utilization of nutritional sources. From this and previous work, we conclude that the ComK regulon defines a growth-arrested state, distinct from sporulation, of which competence for genetic transformation is but one notable feature. We suggest that this is a unique adaptation to stress and that it be termed the 'K-state'. PMID- 11918819 TI - Bacterial secrets of secretion: EuroConference on the biology of type IV secretion processes. AB - Type IV secretion systems (TFSS) mediate secretion or direct cell-to-cell transfer of virulence factors (proteins or protein-DNA complexes) from many Gram negative animal, human and plant pathogens, such as Agrobacterium tumefaciens, Bartonella tribocorum, Bordetella pertussis, Brucella suis, Helicobacter pylori, Legionella pneumophila and Rickettsia prowazekii, into eukaryotic cells. Bacterial conjugation is also classified as a TFSS-like process mediating the spread of broad-host-range plasmids between Gram-negative bacteria such as RP4 and R388, which carry antibiotic resistance genes. Genetic, biochemical, cell biological and structural biology experiments led to significant progress in the understanding of several aspects of TFSS processes. X-ray crystallography revealed that homologues of the A. tumefaciens inner membrane-associated proteins VirB11 and VirD4 from H. pylori and R388, respectively, may form channels for substrate translocation or assembly of the transmembrane TFSS machinery. Biochemical and cell biological experiments revealed interactions between components of the periplasmic core components VirB8, VirB9 and VirB10, which may form the translocation channel. Analysis of A. tumefaciens virulence proteins VirE2 and VirF suggested that the periplasmic translocation route of the pertussis toxin from B. pertussis may be more generally valid than previously anticipated. Secretion and modification of toxins from H. pylori and L. pneumophila profoundly affect host cell metabolism, thus entering the discipline of cellular microbiology. Finally, results from genome sequencing projects revealed the presence of up to three TFSS in a single organism, and the analysis of their interplay and adaptation to different functions will be a future challenge. TFSS-carrying plasmids were discovered in different ecosystems, suggesting that genetic exchange may speed up their evolution and adaptation to different cell-cell interactions. PMID- 11918818 TI - The switch from inorganic to organic sulphur assimilation in Escherichia coli: adenosine 5'-phosphosulphate (APS) as a signalling molecule for sulphate excess. AB - The utilization of organosulphur compounds as sources of sulphur by Escherichia coli is strongly repressed by sulphate. To search for the signal enabling E. coli to alternate gene expression according to the sulphur source, we investigated the transcriptional control of the ssuEADCB operon, required for the transport and desulphonation of aliphatic sulphonates. We demonstrate that, of the two LysR type regulators involved in expression from the ssu promoter, Cbl acts as a direct and sufficient activator of transcription in vivo and in vitro, whereas CysB downregulates the promoter efficiency. Most importantly, the Cbl-mediated transcription initiation at the ssu promoter in vitro is abolished in the presence of an early metabolite of the sulphate assimilatory pathway, adenosine 5'-phosphosulphate (APS). This role for APS was confirmed in vivo by measuring the expression of beta-galactosidase from a transcriptional ssu-lacZ fusion in strains containing different mutations blocking the synthesis and consumption of APS. Our data comprise the first evidence that APS may act as the negative cofactor of the transcriptional regulator Cbl, and that APS, and not sulphate itself, serves as the signalling molecule for sulphate excess. PMID- 11918820 TI - Use of economic evaluation in decision making: what needs to change? PMID- 11918821 TI - The bias against new innovations in health care: value uncertainty and willingness to pay. AB - This paper offers a model for the bias found in willingness-to-pay valuations against new treatments. For example, this bias provides an explanation for patient preferences that make it difficult for formularies to take treatments off their lists, even when newer treatments would appear to be clearly preferable. The appeal of the model, which is based on imperfect information, is that it is consistent with rational preferences and rational behavior by patients, which are necessary for standard models and methods related to decision theory, cost effectiveness, and efficiency. PMID- 11918822 TI - Do health-care decision makers find economic evaluations useful? The findings of focus group research in UK health authorities. AB - OBJECTIVES: The impact of economic evaluation studies on health-care decision makers has been shown to be rather limited. However, there is an increasing requirement for the cost-effectiveness of health-care interventions to be considered in formulating and implementing guidelines for clinical practice. This paper reports the findings of recent focus group research among UK health authorities, which examined the usefulness of published economic evaluations within the decision-making processes. The findings are presented and discussed in light of other studies that have addressed this issue. METHODS: Focus group research was conducted with decision makers from a sample of two UK health authorities using the National Health Service Economic Evaluation Database (NHS EED) as a research vehicle to locate and report the findings of relevant economic studies. The study sample was initially invited to respond to questionnaires exploring the usefulness of published economic evaluations in the decision-making process and to outline particular topics that it felt would benefit from similar economic evidence. Following this, a detailed search was undertaken to retrieve structured NHS EED abstracts on these topics such that the usefulness and limitations of economic evaluations to decision making could be determined. RESULTS: Decision makers generally recognized the usefulness and necessity of published economic evaluations in informing their decision-making processes. However, the value of studies was often limited because of the poor generalizability of results, the narrowness of research questions, and the lack of methodological rigor common to many published studies. A total of 237 NHS EED full abstracts were retrieved in the specified areas of interest, which, within specified caveats, were generally found to be useful as decision-making tools. There was a general consensus among decision makers in favor of developing a quality-scoring system for studies, thereby going beyond the critical summaries given in NHS EED. CONCLUSIONS: Decision makers value information on cost effectiveness as well as effectiveness alone, but methodological improvements are necessary to increase the reliability of economic studies. A quality-scoring system for published studies would be a useful development as a filtering mechanism for decision makers but would raise a number of challenges for health economists. PMID- 11918823 TI - Is sepsis accurately coded on hospital bills? AB - OBJECTIVE: To examine whether sepsis is accurately coded on hospital bills. METHODS: Hospital inpatient uniform bills (UB-92) for 122 patients with clinically documented severe sepsis of presumed infectious origin were retrospectively examined. Final UB-92 hospital bills were obtained for all study subjects. ICD-9-CM diagnosis codes from these bills were then reviewed to ascertain the number of subjects for whom one or more diagnostic codes for septicemia and/or bacteremia were present. RESULTS: A total of 92 hospital bills (75.4%) contained one or more ICD-9-CM diagnostic codes for septicemia and/or bacteremia. Of the 30 that did not, 15 (12.3%) had codes for major systemic infection and organ failure. No diagnoses indicative of sepsis (i.e., organ failure and major infection) were present on the remaining 15 (12.3%) bills. CONCLUSIONS: Our findings suggest that use of ICD-9-CM codes for identifying patients with sepsis using hospital bills is only moderately sensitive. Strict reliance on administrative data sources for sepsis surveillance or research planning may therefore be prone to substantial error. PMID- 11918824 TI - Development of a decision-analytic model of stroke care in the United States and Europe. AB - OBJECTIVE: Stroke places a huge burden on society in terms of premature death, disability, and costs of care. Increasingly, the cost-effectiveness of new interventions needs to be demonstrated before their widespread implementation. Clinical trials are unable to measure the long-term impact of such new interventions in stroke care, and a modeling approach is necessary. The Stroke Outcome Model has been developed in four countries: France, Germany, the United Kingdom, and the United States as a flexible tool for this purpose. METHOD: The decision-analytic model represents the management of acute stroke and long-term care and prevention of recurrence for stroke survivors. The latter consists of semi-Markov state-transition processes, with health states defined by therapy, disability, and occurrence of further stroke. Sources of clinical data include trials, meta-analyses, and prospective cohort studies such as the Oxfordshire Community Stroke Project and the Northern Manhattan Stroke Study. Resource use data were obtained from published sources and expert clinician panels. Outcome measures used were strokes averted, life years, and quality-adjusted life-years gained. RESULTS: The model has been used to undertake economic analyses of antiplatelet therapy for the prevention of recurrent strokes, and of stroke unit care and thrombolytic therapy in acute stroke. From a health- and social-care perspective, new interventions were found to be cost saving or to provide health benefits at modest additional cost. Results were sensitive to the cost perspective, time horizon, baseline risk of stroke recurrence, and choice of effectiveness measure. CONCLUSION: The development of this model highlights the need for improved information on prognosis and resources used by stroke survivors and the importance of differentiating between economically distinct end points such as death, disabled survival and nondisabled survival, which may be combined as outcomes in clinical trials. PMID- 11918825 TI - Automated current health time-trade-off assessments in women's health. AB - OBJECTIVE: To motivate the role for preference assessment in women's health and to report pilot data addressing the performance of automated time-trade-off (TTO) valuations of current health, which were developed to estimate quality-adjusted life years (QALYs) in the women's health setting. METHODS: Values for current health relative to perfect health and death were assessed using an annual time trade-off (1-year horizon and sleep as the trading metaphor), a lifetime time trade-off, and a visual analog scale (VAS). All instruments were administered twice within a 12- to 14-day window among a convenience sample of 27 women. RESULTS: Valuation of health was similar for both time trade-offs (mean of 0.95 for both), but was significantly lower for the VAS (mean of 0.84, Wilcoxon signed rank p-value < 0.001). Reliability using the intraclass correlation coefficient was 0.67 +/- 0.09 and 0.75 +/- 0.07 for the annual and lifetime time trade-offs, respectively, and 0.89 +/- 0.03 for the VAS. Construct validity was supported by consistent trends in time-trade-off utilities across tertiles of the Short Form 36 (SF-36) general health subscale (trend test p-value < 0.001). CONCLUSION: Automated time trade-offs for current health provide a promising approach for use in women's health studies where impact on QALYs must be measured. Natural areas of application include the economic evaluation of preventive interventions in postmenopausal women. PMID- 11918826 TI - Validity of a Work Productivity and Activity Impairment questionnaire for patients with symptoms of gastro-esophageal reflux disease (WPAI-GERD)--results from a cross-sectional study. AB - OBJECTIVES: To validate a Work Productivity and Activity Impairment questionnaire (WPAI-GERD) developed to measure lost productivity due to symptoms of gastro esophageal reflux disease (GERD). METHODS AND DATA: The WPAI-GERD was administered along with two quality-of-life questionnaires, Quality of Life in Reflux and Dyspepsia (QOLRAD) and Short Form 36 (SF-36), to a Swedish working population (N = 136) visiting a general practitioner for symptoms attributed to GERD. Correlation coefficients were calculated between each productivity variable derived from the WPAI-GERD and symptom severity, symptom frequency, quality of life dimensions, age, and gender. Statistical tests were carried out to determine the relationship between each productivity variable and the severity of heartburn. RESULTS: High correlations (range: 0.30-0.75) were found between productivity and symptom severity as well as between productivity and quality-of life dimensions related to work and daily activities. The results demonstrated the ability of the questionnaire to discriminate between different grades of heartburn severity. On average, patients with heartburn reported 2.5 hours absence from work, 23% reduced productivity while at work, and 30% reduced productivity while doing regular daily activities during the week preceding the consultation. The SF-36 scores implied that patients, especially those with moderate-to-severe heartburn, had a poor quality of life compared with a normal population. CONCLUSION: The results indicate a high convergent and discriminant validity of the WPAI-GERD questionnaire and also show that patients consulting a physician because of symptoms attributed to GERD report substantial impairment in both productivity and health-related quality of life. PMID- 11918827 TI - Re: Hutton J, Brown R. Use of economic evaluation in decision-making: what needs to change? (Editorial) Value Health 2002;5:65-6. PMID- 11918828 TI - Bootstrap, Bayesian probability and maximum likelihood mapping: exploring new tools for comparative genome analyses. AB - BACKGROUND: Horizontal gene transfer (HGT) played an important role in shaping microbial genomes. In addition to genes under sporadic selection, HGT also affects housekeeping genes and those involved in information processing, even ribosomal RNA encoding genes. Here we describe tools that provide an assessment and graphic illustration of the mosaic nature of microbial genomes. RESULTS: We adapted the Maximum Likelihood (ML) mapping to the analyses of all detected quartets of orthologous genes found in four genomes. We have automated the assembly and analyses of these quartets of orthologs given the selection of four genomes. We compared the ML-mapping approach to more rigorous Bayesian probability and Bootstrap mapping techniques. The latter two approaches appear to be more conservative than the ML-mapping approach, but qualitatively all three approaches give equivalent results. All three tools were tested on mitochondrial genomes, which presumably were inherited as a single linkage group. CONCLUSIONS: In some instances of interphylum relationships we find nearly equal numbers of quartets strongly supporting the three possible topologies. In contrast, our analyses of genome quartets containing the cyanobacterium Synechocystis sp. indicate that a large part of the cyanobacterial genome is related to that of low GC Gram positives. Other groups that had been suggested as sister groups to the cyanobacteria contain many fewer genes that group with the Synechocystis orthologs. Interdomain comparisons of genome quartets containing the archaeon Halobacterium sp. revealed that Halobacterium sp. shares more genes with Bacteria that live in the same environment than with Bacteria that are more closely related based on rRNA phylogeny. Many of these genes encode proteins involved in substrate transport and metabolism and in information storage and processing. The performed analyses demonstrate that relationships among prokaryotes cannot be accurately depicted by or inferred from the tree-like evolution of a core of rarely transferred genes; rather prokaryotic genomes are mosaics in which different parts have different evolutionary histories. Probability mapping is a valuable tool to explore the mosaic nature of genomes. PMID- 11918830 TI - A theoretical model for estimating the margination constant of leukocytes. AB - BACKGROUND: Blood leukocytes constitute two interchangeable sub-populations, the marginated and circulating pools. These two sub-compartments are found in normal conditions and are potentially affected by non-normal situations, either pathological or physiological. The dynamics between the compartments is governed by rate constants of margination (M) and return to circulation (R). Therefore, estimates of M and R may prove of great importance to a deeper understanding of many conditions. However, there has been a lack of formalism in order to approach such estimates. The few attempts to furnish an estimation of M and R neither rely on clearly stated models that precisely say which rate constant is under estimation nor recognize which factors may influence the estimation. RESULTS: The returning of the blood pools to a steady-state value after a perturbation (e.g., epinephrine injection) was modeled by a second-order differential equation. This equation has two eigenvalues, related to a fast- and to a slow-component of the dynamics. The model makes it possible to identify that these components are partitioned into three constants: R, M and SB; where SB is a time-invariant exit to tissues rate constant. Three examples of the computations are worked and a tentative estimation of R for mouse monocytes is presented. CONCLUSIONS: This study establishes a firm theoretical basis for the estimation of the rate constants of the dynamics between the blood sub-compartments of white cells. It shows, for the first time, that the estimation must also take into account the exit to tissues rate constant, SB. PMID- 11918829 TI - PISA. The effect of paracetamol (acetaminophen) and ibuprofen on body temperature in acute stroke: protocol for a phase II double-blind randomised placebo controlled trial [ISRCTN98608690]. AB - BACKGROUND: During the first days after stroke, one to two fifths of the patients develop fever or subfebrile temperatures. Body temperature is a strong prognostic factor after stroke. Pharmacological reduction of temperature in patients with acute ischaemic stroke may improve their functional outcome. Previously, we studied the effect of high dose (6 g daily) and low dose (3 g daily) paracetamol (acetaminophen) in a randomised placebo-controlled trial of 75 patients with acute ischemic stroke. In the high-dose paracetamol group, mean body temperature at 12 and 24 hours after start of treatment was 0.4 degrees C lower than in the placebo group. The effect of ibuprofen, another potent antipyretic drug, on body core temperature in normothermic patients has not been studied. AIM: The aim of the present trial is to study the effects of high-dose paracetamol and ibuprofen on body temperature in patients with acute ischaemic stroke, and to study the safety of these treatments. DESIGN: Seventy-five (3 x 25) patients with acute ischaemic stroke confined to the anterior circulation will be randomised to treatment with either: 400 mg ibuprofen, 1000 mg acetaminophen, or with placebo 6 times daily during 5 days. Body-temperatures will be measured with a rectal electronic thermometer at the start of treatment and after 24 hours. An infrared tympanic thermometer will be used to monitor body temperature at 2-hour intervals during the first 24 hours and at 12-hour intervals thereafter. The primary outcome measure will be rectal temperature at 24 hours after the start of treatment. The study results will be analysed on an intent-to-treat basis, but an on-treatment analysis will also be performed. No formal interim analysis will be carried out. PMID- 11918831 TI - Effect of taurine on platelets and the plasma coagulation system. AB - It is not yet clear what exact mechanisms are at work in hibernating animals that prevent clot formation and maintain tissue perfusion under conditions of very slow blood flow and increased blood viscosity brought about by the low temperatures. It has been shown that the total amino acid pool increases more then two fold in hibernating animals with taurine accounting for about 50% of this increase [Storey et al., Proc Natl Acad Sci USA 1988; 85(21): 8350-4]. This work investigates the effect of taurine on platelets and the plasma coagulation system. Taurine was added at different concentrations in the range between 5 and 25 mM to donor plasma. Using STA/STA Compact coagulation analyzer the following tests were performed: prothrombin time (PT), activated partial thromboplastin time (APTT), and thrombin time (TT). At the highest concentration tested (25 mM) taurine prolonged TT by 9%. The prolongation was statistically significant but not clinically significant retaining TT within normal limits (16.7-20.7 s). PT and APTT remained unchanged by taurine. The effect of taurine on platelets was assessed by platelet aggregation by thrombin, extent of platelet shape change (ESC) induced by ADP, and thrombelastography. Taurine at 5 mM final concentration inhibited platelet aggregation by 10%. Increasing taurine concentration to 25 mM did not result in a further augmentation of the inhibitory effect. ESC was unaffected by taurine. Clot strength determined by thrombelastography also remained unchanged by taurine. PMID- 11918832 TI - Cold temperatures reduce the sensitivity of stored platelets to disaggregating agents. AB - In this study, we compared the effect of signal transduction inhibitors on fibrinogen binding, aggregation, the activation state of GPIIb-IIIa, and cytosolic calcium levels in cold and room temperature-stored platelets. Cold stored platelets have a higher sensitivity to agonist-induced aggregation when compared to room temperature-stored platelets. We also found that cold-stored platelets had a significantly higher aggregation response to ADP and epinephrine, while platelets stored at room temperature responded poorly to these agonists (mean values of 61 vs. 18%, n = 14). Four inhibitors were selected to target various signaling pathways. Cold-stored platelets were more resistant to disaggregation by promethazine, prostaglandin D2, yohimbine, and echistatin. The effects of cold temperatures on stored platelets are targeted to activation pathways as there was no spontaneous aggregation or spontaneous fibrinogen binding as measured in this study. PAC-1 binding was not inhibited to the same degree as aggregation or fibrinogen binding responses, suggesting that the disaggregation was not caused by a change in the conformation of GPIIb-IIIa. Cytosolic calcium levels did not decrease in cold-stored platelets after inhibitor addition. The inhibitors are likely acting after the establishment of the GPIIb-IIIa activation state and may affect the post-occupancy signaling by the fibrinogen-occupied integrin. Differences between aggregation and disaggregation responses of cold- and room temperature-stored platelets suggest that cold-stored platelets may have different mechanisms to stabilize platelet aggregates during their formation. PMID- 11918833 TI - Aggregation of human platelets by gingipain-R from Porphyromonas gingivalis cells and membrane vesicles. AB - The hypothesis that there is an association between periodontitis and cardiovascular disease suggests new lines of research on the mechanism whereby oral bacteria might exert systemic effects. This study was conducted to ascertain and quantitate the effect of Porphyromonas gingivalis on human platelets in vitro. A second related objective was to purify and identify the aggregating vector. Aggregation was measured by platelet turbidometry and gingipain-R was purified from P. gingivalis membrane vesicles by Sepharose 2B and hydroxyapatite chromatography. The in vitro aggregation of platelets requires that at least 1.0 x 10(4) cells be stirred with 1.35 x 10(8) platelets. The specific activity is substantially increased in the membrane vesicles that are shed by this bacterium. Aggregability was due to gingipain-R activity, a potent cysteine protease that was found to be highly concentrated in the membrane vesicle fraction. The enzyme was purified 18-fold in high yield from the membrane vesicles, and consists of two noncovalently linked proteins that migrate at 49 and 44 kDa on SDS-PAGE. Aggregation of platelets by gingipain-R was shown to be dose-dependent, and inhibited by leupeptin and arginine, but not by anti-thrombin III. This is the first report enumerating the specific number of cells and lowest concentration of membrane vesicles necessary to evoke a full human platelet response, and the first report to assign this activity to gingipain-R. PMID- 11918834 TI - Acyl specificity of phospholipases A2 and C in thrombin-stimulated human platelets. AB - Platelets were labelled separately with six different, radioactive unsaturated fatty acids. The cells were isolated from the radioactive precursors and treated with and without 2 U/ml of thrombin. The formation of radioactive free fatty acid+oxygenated fatty acids and of radioactive radioactive phosphatidic acid+diacylglycerol was taken as a measure of the PLA(2) and PLC reactions, respectively. We found that that in intact platelets PLA(2) prefers phospholipid molecular species containing unsaturated acyls, most likely in the sn-2 position, in the priority order: 20:4>20:5>18:2 = 18:3 = 22:6>>18:1, while PLC prefers its substrates in the priority order 20:5>20:4>18:2>18:3 = 22:6>18:1. PMID- 11918835 TI - Towards a definition of aspirin resistance: a typological approach. AB - 'Aspirin resistance' is a poorly defined term to describe the inability of aspirin to protect individuals from thrombotic complications and there are conflicting reports on incidence rates and clinical relevance of this phenomenon. Using collagen (1 microg/ml)-induced platelet aggregation and thromboxane formation (measured as thromboxane B(2)) in citrated platelet-rich plasma, this study demonstrates that aspirin resistance can be classified into three distinct types. In aspirin responders, both, collagen-induced platelet aggregation and thromboxane formation was completely (>95%) inhibited by oral aspirin treatment (100 mg/day). In type I resistance (pharmacokinetic type), oral treatment with aspirin was ineffective but addition of aspirin (100 microM) in vitro resulted in a complete inhibition of collagen-induced platelet aggregation and thromboxane formation. In type II resistance (pharmacodynamic type), neither oral treatment with aspirin nor addition of aspirin in vitro inhibited collagen-induced platelet aggregation and thromboxane formation. In type III resistance (pseudo resistance), platelet aggregation was induced by a low concentration of collagen (1 microg/ml) despite of a complete inhibition of thromboxane formation by oral aspirin treatment. This typology of aspirin resistance should help to clarify the mechanisms, the actual rate, and the possible clinical consequences of this phenomenon. PMID- 11918837 TI - Molecular cloning of a cDNA encoding a bovine growth differentiation factor-9 (GDF-9) and expression of GDF-9 in bovine ovarian oocytes and in vitro-produced embryos. AB - Growth differentiation factor-9 (GDF-9), a member of the transforming growth factor-beta superfamily, is known to be expressed specifically in ovaries of various mammalian species and to be important for normal follicular development in mice. In the present study, a cDNA encoding for bovine GDF-9 was isolated and characterized, and expression of GDF-9 mRNA in ovarian oocytes and in-vitro derived embryos was examined. Isolation of bovine GDF-9 was achieved using the polymerase chain reaction (PCR) with primers based on an ovine GDF-9 cDNA sequence. A 1385 bp cDNA encodes a deduced 453-amino acid sequence which contains the proregion (318 residues) and the mature protein (135 residues) portion. The deduced amino acid sequence of bovine GDF-9 is 98 and 93% identical to ovine GDF 9 and human GDF-9 in the mature portion of the molecule, respectively. Results from reverse transcription (RT)-PCR analysis detected bovine GDF-9 mRNA in preantral follicles (150-200 microm in diameter), early antral follicles (400-800 microm in diameter), and immature oocytes, whereas no detectable PCR signal was observed in cumulus/granulosa cells. In addition, bovine GDF-9 mRNA continued to be expressed in developing embryos up to the 8-cell stage, but was undetectable at the blastocyst stage. These findings give rise to new possibilities regarding an additional physiological role of GDF-9 in early embryonic development as well as in the development of follicles. PMID- 11918838 TI - Lipid composition of virosomes modulates their fusion efficiency with cryopreserved bull sperm cells. AB - Virosomes derived from different fusogenic enveloped viruses have been generated for potential application in gene targeting to sperm cells. Comparative characterization of reconstitution products revealed that virosomes derived from influenza viruses are superior to those generated from Sendai viruses, with respect to the fusion rates with cryopreserved bull sperm cells and to sperm cell vitality after fusion. Modulation of the lipid composition during virosome reconstitution affects fusion sites on target sperms and allows optimization of the fusion rate and sperm cell vitality. A fluorescence-based microscopic fusion assay combined with a vital cell stain revealed that about 90% of sperm cells fused with influenza virosomes containing exogenous cholesterol, sphingomyelin, phosphatidylcholine, and phosphatidylethanolamine. About 85% of the fused sperm cells remained vital. Such optimized influenza-derived virosomes provide the basis for ongoing experiments, which aim at eventually generating biologically active transgenic sperms. PMID- 11918839 TI - Preimplantation screening for transgenesis using an embryonic specific promoter and green fluorescent protein. AB - We report enrichment in the efficiency of generating mice transgenic for expression of a human protein in their milk using GFP-mediated preimplantation screening. The transgene array consisted of a functional gene (human alpha-1 antitrypsin under the control of the ovine BLG promoter) linked 5' to a reporter gene (GFP under the control of the murine Oct-4 promoter). GFP expression was detected in blastocysts by fluorescence microscopy and green and nongreen embryos were transferred to recipients in separate groups. In the first experiment, of seven pups that resulted from the transfer of blastocysts expressing GFP, five (71%) were transgenic. The experiment was repeated and of 12 pups that resulted from transfer of GFP-expressing blastocysts, 11 were transgenic (92%). The presence of the reporter cassette used for preimplantation screening did not affect the expression level of alpha-1-antitrypsin in the milk of the transgenic mice. In addition, in a related experiment wherein the GFP reporter gene was co injected with a second mammary-specific transgene, pINC, no effect on transgene expression was observed. For mice transgenic for the mammary-specific gene alone, expression levels for four different lines were 192, 197, 382, and 415 microg/mL. For mice transgenic for both the mammary-specific transgene and the Oct4-GFP reporter cassette, expression levels for seven different lines were 282, 321, 468, 497, 499, 516, and 806 microg/mL. PMID- 11918840 TI - Production of cloned goats from enucleated oocytes injected with cumulus cell nuclei or fused with cumulus cells. AB - This study was designed to produce cloned goats from cumulus cells. Cloning donor nuclei were from cumulus cells either freshly isolated or cultured in vitro. Enucleated oocytes were either injected with cumulus cell nuclei without piezo driven manipulator (injection method) or fused with cumulus cells (fusion method). The survival rate of cloned embryos, obtained by injection, was higher than that derived from fusion (62.7 and 45.9%, respectively). Two cloned goats were derived by fusion with in vitro cultured cumulus cells without starvation, but died shortly after natural birth, from respiratory difficulties. Their birth weights (2.23 kg and 2.03 kg) were within the normal range (2.0-2.7 kg) and postmortem analysis revealed no morphological abnormalities. The third cloned goat, derived by injection of nuclei from freshly isolated cumulus cells, weighed 3.3 kg at birth, and was 37% overweight compared with the average weight of the same species. This goat is healthy and well as this paper is being prepared. Nested PCR-RFLP analysis confirmed that all the cloned goats were derived from the donor cells. PMID- 11918841 TI - Potential food contaminants and associated health risks. AB - The potential toxicants in food are derived from natural or industrial sources. Compounds like lectins and glycoalkaloids that are toxic to man are naturally present in some vegetables like potatoes or legumes. A wide variety of marine toxins mostly produced by dinoflagellates occurring secondarily in molluscs and mussels are usually ingested by human beings causing poisoning. On the other hand, toxic compounds find their way into food during manufacture, storage, or transportation. These include largely the industrial contaminants, persistent organic pollutants (POP), pesticides, heavy metals, and toxins of fungal and bacterial origin. Further, toxic compounds like higher alcohols may be produced as byproducts during processing. Migration of compounds from packaging materials into packaged food like contamination with lead from solder in certain metal cans is well known. Additives (emulsifiers, preservatives, and antioxidants) could also influence the quality of foods. Solvent residues may find their way into food as a result of their use in extraction processes like the use of trichloroethylene and methylene chloride in decaffeination of coffee. In addition, poor hygiene, storage, and preparation may also lead to food contamination by various microbes and ova or cysts of nematodes. The problem of food contamination can be overcome to a great extent by regular surveillance and monitoring programmes and strict implementation of food and adulteration act. In the present review some of these aspects of food contamination have been discussed in detail. PMID- 11918842 TI - Kininase-independent potentiation of endothelium-dependent relaxations to kinins by converting enzyme inhibitor perindoprilat. AB - AIM: The present study examined whether or not the resistance to degradation of bradykinin analogs affects the kinin-potentiating action of the inhibitor of converting enzyme, perindoprilat. METHODS: Hydrolysis of Hyp3,Tyr(Me)8 bradykinin by ACE present in isolated canine coronary arteries was assessed by determination of peptide metabolites using electrospray mass spectrometry, and compared to that of bradykinin. Contractions and relaxations of isolated rings of coronary arteries, with and without endothelium, were recorded as changes in isometric force. RESULTS: After a 30 min incubation, most of the bradykinin was degraded by the arteries, while less than 10 % of Hyp3,Tyr(Me)8 -bradykinin was hydrolysed. In organ chambers, Hyp3,Tyr(Me)8 -bradykinin like bradykinin caused relaxations of isolated canine coronary arteries with endothelium that could be attributed to both NO and endothelium-derived hyperpolarizing factor (EDHF). Perindoprilat caused a comparable leftward shift in the concentration-relaxation curves for bradykinin and Hyp3,Tyr(Me)8 -bradykinin. CONCLUSION: Impairment of the degradation of bradykinin is not an essential mechanism by which perindoprilat potentiates the actions of kinins. PMID- 11918843 TI - Molecular simulation of interaction between estrogen receptor and selective estrogen receptor modulators. AB - AIM: To study the mechanism of interaction between a series of potent racemic selective estrogen receptor modulators (SERM) and estrogen receptors (ER). METHODS: Active conformations of these conformationally restricted raloxifene analogues in binding pocket were determined by molecular mechanics. The interactive energies between ligand and receptor were calculated by docking program. RESULTS: Both R and S configurations of these SERM were accommodated by the binding pocket of ER. The hydroxy group of compounds forms hydrogen bonds with amino acid residues of ER and the phenolic group mimics the A ring of estradiol. The most potential compounds were those with two hydroxy groups and accommodated by binding pocket in S configuration with phenolic group at C(16) imitating A ring of estradiol. CONCLUSION: Chiral center conferred little effect on the binding affinity of these conformationally restricted raloxifene analogues. The hydroxy group(s) play(s) a critical role to the orientation of compounds in active pocket of ER and the binding between ligand and receptor. PMID- 11918844 TI - Estrogen prevents bovine aortic endothelial cells from TNF-alpha-induced apoptosis via opposing effects on p38 and p44/42 CCDPK. AB - AIM: To investigate the effect of 17beta-estradiol (E2) on TNF-alpha-induced apoptosis in cultured bovine aortic endothelial cells (BAEC) and the underlied mechanism. METHODS: BAEC were cultured and passaged in Dulbecco's modified Eagle's medium (DMEM). Morphologic changes and quantification of apoptotic cells were determined under fluorescence microscope with Hoechst 33258 staining. Cell viability was detected with MTT method. DNA fragmentation was visualized by agarose gel electrophoresis. The expression of phospho-p38 and phospho-p44/42 Ca2+-calmodulin dependent protein kinase (CCDPK) was measured by Western blotting. RESULTS: TNF-alpha 5000 kU/L elicited typical apoptotic morphologic changes (chromatic condensation, nucleus fragmentation, and DNA fragmentation). E2 0.1 pmol/L-100 nmol/L enhanced the expression of phospho-p44/42 CCDPK induced by TNF-alpha, at the same time, inhibited TNF-alpha induced activation of p38 CCDPK. E2 protected BAEC from apoptosis induced by TNF-alpha in a concentration dependent manner. DNA fragmentation induced by TNF-alpha 5000 kU/L was also reduced by E2 1 nmol/L. Both the E2-induced upregulation of phospho-p44/42 CCDPK and its anti-apoptotic action were prevented by the specific p44/42 CCDPK inhibitor U0126. CONCLUSION: Activation of p44/42 CCDPK signaling together with inhibition of p38 CCDPK signaling by E2 appears to be an important mechanism for its survival effect on endothelial cells. PMID- 11918845 TI - Effect of agmatine on L-type calcium current in rat ventricular myocytes. AB - AIM: To study the effect of agmatine (Agm) on L-type calcium current (I(Ca-L)) in rat ventricular myocytes. METHODS: Whole-cell configuration of the patch-clamp technique was used to record I(Ca-L) in single rat ventricular myocytes which were dissociated by enzymatic dissociation method. RESULTS: (1) Agm (0.5, 1, 2 mmol/L) reduced the voltage-dependently activated peak amplitude of I(Ca-L) (pA) from 1451+/-236 (control) to 937+/-105 (n=8, P <0.05), 585+/-74 (n=8, P <0.01), and to 301+/-156 (n=8, P <0.01) in a concentration-dependent manner. (2) Agm (1 mmol/L) blocked I(Ca-L) in a use-dependent manner. The degree of use-dependent blocking effect was 53 %+/-12 % (n=8, P <0.05) at 1 Hz, and 69 %+/-11 % (n=8, P <0.01) at 3 Hz. (3) Agm upshifted the current-voltage (I-V) curve, but the characteristics of I-V relationship were not significantly altered by Agm, the maximal activation voltage of I(Ca-L) was not different from that of control. Steady-state activation of I(Ca-L) was not affected markedly. The half activation potential (V(0.5)) and the slope factor (k) were not significantly different from those of the control. V(0.5) value was (-20.2+/-2.5) mV in the control and ( 20.5+/-2.7) mV in the presence of Agm 1 mmol/L. The k value was (7.1+/-0.4) mV and (7.5+/-0.5) mV, respectively (n=8, P >0.05). (4) Agm 1 mmol/L markedly shifted the steady-state inactivation curve of I(Ca-L) to the left, and accelerated the voltage-dependent steady-state inactivation of calcium current. V(0.5) value was (-32+/-6) mV in the control and (-40+/-5) mV in the presence of Agm. The k value was (7.6+/-0.9) mV and (12.5+/-1.1) mV, respectively (n=8, P <0.05). (5) Agm 1 mmol/L markedly delayed half-recovery time of Ca2+ channel from inactivation (92+/-28) ms to (249+/-26) ms (n=8, P <0.01). CONCLUSION: Agm inhibited I(Ca-L) and mainly acted on the inactivated state of L-type calcium channel, manifested as acceleration of calcium channel inactivation and slowdown of recovery from inactivated state in rat ventricular myocytes. PMID- 11918846 TI - Cyclosporin A enhanced protection of nimodipine against brain damage induced by hypoxia-ischemia in mice and rats. AB - AIM: To study whether P-glycoprotein (P-gp) inhibitor cyclosporin A (CsA) enhanced the protection of nimodipine (NMD) against brain damage. METHOD: (1) After mice were given ip NMD alone or co-administration of CsA, survival time of mice were recorded following decapitation and ip injection of NaNO2, respectively. (2) After rats were given ip NMD alone or co-administration of CsA, 20 min forebrain ischemia induced by the technique of two-carotid occlusion plus hypovolemic hypotension. Following reperfusion of 1 h, the content of malondialdehyde (MDA), lactic acid (LA) and activity of lactic dehydrogenase (LDH) in cortex tissue were measured. (3) NMD level in brain was also determined after ip injection of NMD 2 mg/kg alone and co-administration of CsA, respectively. RESULTS: NMD showed potent pharmacological activity in the three models. The survival time of mice by decapitation and ip NaNO2 were significantly (P <0.05) prolonged after co-administration of CsA. In rat forebrain ischemia/reperfusion, levels of MDA, LA, and LDH by co-administration of CsA were greatly modified, compared with those of NMD alone group. The level of NMD in rat brain was increased markedly after co-administration of CsA. CONCLUSION: P-gp inhibitor CsA may enhance the protection of NMD against brain damage. PMID- 11918847 TI - Effects of various muscarinic ligands on M2AChR-Gi1alpha fusion protein expressed in Sf9 insect cells. AB - AIM: To generate M2AChR-Gi1alpha fusion protein in baculovirus-Sf9 cells system and detect the effects of various muscarinic ligands and magnesium ion on the interaction of fused M2AChR and Gi1alpha. METHODS: M2AChR-Gi1alpha fused DNA was generated in a two step polymerase chain reaction (PCR) and then expressed in Sf9 cells to produce fusion protein. [3H] L-quinuclidinyl benzilate (QNB) and 35S GTPgammaS binding experiments were performed to study the function of M2AChR Gi1alpha fusion protein. RESULTS: The expression level of M2AChR-Gi1alpha was (20.12 +/- 0.14) nmol/g protein. The affinity of GDP to Gi1alpha partner changed in the presence of different muscarinic ligands. IC50 values (95 % confidence limit) of GDP in the presence of acetylcholine (ACh), carbamylcholine, (4-hydroxy 2-butynyl)-1-trimethylammonium-m-chloro-carbanilate chloride (McN-A-343), pilocarpine, and atropine were 178 (148 - 214) micromol/L, 158 (126 - 199) micromol/L, 66 (56 - 78) micromol/L, 62 (55 -7 2) micromol/L, and 5.0 (4.6 - 5.5) micromol/L, respectively, and that in the absence of muscarinic ligand was 15.9 (14.3 - 17.6) micromol/L. Apparent affinity for GDP in the presence of carbamylcholine was markedly decreased with increasing MgCl2 concentrations, although the apparent affinity in the presence of atropine was not affected. CONCLUSION: The M2AChR-Gi1alpha fusion protein has the pharmacological specificity of M2 receptor and the efficient signaling of the two partners. Affinity of GDP to ligand-bound fusion protein represents the species of muscarinic ligands. Mg2+ is necessary for the action of M2AChR on Gi1alpha. PMID- 11918848 TI - Modulation of neuronal nicotinic acetylcholine receptors by glucocorticoids. AB - AIM: To investigate the nongenomic effect of the glucocorticoid corticosterone on nicotinic acetylcholine receptor (nAChR) in PC12 cells. METHODS: The acetylcholine (ACh)-induced current was measured on nerve growth factor differentiated PC12 cells using whole-cell patch-clamp techniques. RESULTS: The ACh-induced current (IACh) proved to be generated through neuronal nAChR. When ACh (30 micromol/L) was applied simultaneously with corticosterone (0.1 - 10 micromol/L), the decay of IACh was faster with slight inhibition on the peak current amplitude. Pretreating PC12 cells with corticosterone augmented the inhibition on the peak IACh and did not alter receptor desensitization. Bovine serum albumin-conjugated corticosterone (0.1 - 10 micromol/L) had the inhibition similar to corticosterone. The rapid effect induced by corticosterone was reversible, concentration-dependent, and voltage-independent. CONCLUSION: Corticosterone has rapid inhibitory effect on IACh, which is mediated by a nongenomic mechanism. It indicates that corticosterone binds to the specific site on the outer cell membrane, probably on the neuronal nicotinic receptor-coupled channel, and inhibits the IACh in a noncompetitive manner, thus controlling the immediate catecholamine release from the sympathetic cells. PMID- 11918849 TI - Effects of cyclosporin A by aerosol on airway hyperresponsiveness and inflammation in guinea pigs. AB - AIM: To study cyclosporin A (CsA) by aerosol for anti-asthmatic effects in guinea pigs. METHODS: PC200 changes of lung resistance (RL) in the antigen-challenged sensitized guinea pig induced by acetylcholine (ACh) or histamine, and eosinophils changes in bronchoalveolar lavage fluid (BALF) and pulmonary histologic section induced by antigen in vivo in sensitized guinea pigs were investigated. RESULTS: Pretreatment with CsA 10 g/L and 20 g/L by aerosol but not with CsA 5 g/L, dexamethasone (DXM) 0.5 mg/kg by ip increased PC200 value and prevented ACh or histamine-induced airway hyperresponsiveness. However, CsA 5 g/L also prevented histamine-induced airway hyperresponsiveness. CsA 10 g/L, 20 g/L and DXM 0.5 mg/kg reduced markedly eosinophil accumulation in BALF. The lymphocyte accumulation induced by antigen was not changed significantly by CsA and DXM tested. DXM 0.5 mg/kg increased number of neutrophil in the BALF. There was a statistical significance comparison with CsA groups. In the pulmonary histological studies, CsA 20 g/L and DXM 0.5 mg/kg also inhibited eosinophil infiltration in the epithelium and subepithelial connective tissue of bronchi and bronchioles. CONCLUSION: Anti-inflammation and anti-hyperresponsiveness of CsA by aerosol in animal model offered an experimental evidence for airway inhalation of CsA in the treatment of asthma. PMID- 11918851 TI - Effects of lidamycin on genomic DNA in human hepatoma BEL-7402 cells. AB - AIM: To further study the effect of enediyne antibiotic lidamycin (C1027) on genomic DNA in human hepatoma BEL-7402 cells. METHODS: The DNA patterns were detected by agarose gel electrophoresis. The gene damage was revealed by Southern hybridization. The DNA repair was determined by neutral agarose gel electrophoresis. The lidamycin-cleaved sites were determined with superhelix GEM plasmid. RESULTS: The DNA ladder patterns were observed after the BEL-7402 cells were treated with lidamycin 1 micromol/L for 15, 30, 60 min. The extrons and introns of active N-ras gene in the BEL-7402 cells were easily damaged by lidamycin at low concentrations, but no damage in the silent IL-2 gene was observed. It was difficult to repair DNA breaks after the BEL-7402 cells were treated with lidamycin. The cleaved sites of GEM plasmid treated with lidamycin were -OH HO-. CONCLUSION: Lidamycin can significantly damage genomic DNA in the hepatoma BEL-7402 cells. The results make it helpful to elucidate the molecular mechanism of potent cytotoxicities of lidamycin to tumor cells. PMID- 11918850 TI - Correlative changes of interferon-gamma and interleukin-4 between cortical layer and pulmonary airway of sensitized rats. AB - AIM: To explore correlation of T helper 1 (Th1) and Th2 cytokine between the cortical layer and pulmonary airway in ovalbumin-induced rat asthma model. METHODS: Aerosol antigen-induced changes of inflammation in bronchoalveolar lavage fluids (BALF) and pulmonary histologic section in sensitized rats were investigated. Changes of interferon-gamma (IFN-gamma) and interleukin-4 (IL-4) in BALF and cortical layer homogenates were determined by ELISA. RESULTS: The number of inflammatory cells in BALF in antigen challenged group was significantly higher than control group (P <0.05). Dexamethasone (DXM, 0.5 mg/kg, ip) markedly reduce total leukocyte numbers in BALF, and almost completely inhibited eosinophil, lymphocyte accumulation. The score of histological examination (eosinophil infiltration, mucous edema and epithelial lesion) in antigen challenged group was significantly higher than control group (P <0.05). DXM (0.5 mg/kg, ip) reduced the numbers of eosinophils and improved mucous edema and epithelial lesion of bronchi and bronchioles. In addition, our results demonstrated decreased IFN-gamma level accompanied by increased IL-4 level that resulted in a decreased IFN-gamma/IL-4 ratio in the BALF after the sensitized rats were challenged with aerosol antigen. Meanwhile, there was a similar change of IFN-gamma and IL-4 in cortical layer homogenates. DXM (0.5 mg/kg, ip) could reverse IFN-gamma/IL-4 ratio in the BALF and cortical layer homogenates in the model. CONCLUSION: The results showed a significant interplay changes of Th1 and Th2 cytokine between central nervous system and pulmonary airway in the asthmatic inflammatory model. PMID- 11918852 TI - Effects of reactive oxygen species on lymphokine-activated killer cells in patients with bladder cancer. AB - AIM: To investigate the effect of reactive oxygen species on the proliferation of lymphokine-activated killer (LAK) cells in patients with bladder cancer and their cytolysis to bladder tumor cells. METHODS: Sodium nitroprusside (SNP) was used as nitric oxide (NO) donor. The superoxide anion (O2-.) was generated in the complete medium (CM) supplemented with N-methylphenazonium methyl sulfate (PMS) 3 120 micromol/L and nicotinamide adenine dinucleotide (NADH) 18 - 600 micromol/L. The hydroxyl radical (.OH) was produced by adding ascorbic acid (AA) 0.5 - 400 micromol/L and ferrous sulfate (Fe2+) 0.05 - 40 micromol/L in CM. LAK cell proliferation and cytotoxicity were assayed in the presence of NO, .OH, or O2-. Bladder cancer cell lines BIU-87 and EJ were cultured as target cells and cytotoxicity of LAK cells were determined by MTT assay. RESULTS: The proliferation of LAK cells induced by interleukin-2 (IL-2) was inhibited by hydroxyl radical from 48 h to 96 h in a dose-dependent fashion and was inhibited to 34.5 % compared with control at 96 h in the concentration of ascorbic acid 400 micromol/L and ferrous sulfate 40 micromol/L. The inhibition induced by.OH can be overcome by certain concentrations of mannitol or editic acid. On the contrary, the proliferation of LAK cells induced by IL-2 was stimulated by certain concentrations of NO or O2-. The stimulation induced by O2-. can be overcome to control level by superoxide dismutase (SOD) 3 10(5) U/L. Exogenous O2-. resulted in an increase in cytotoxicity of LAK cells against BIU87 and EJ cells. However, the LAK cells cytotoxicity treated with hydroxyl radical or SOD showed no difference as compared with the control. CONCLUSION: NO and O2-. enhanced the proliferation and activation and O2-. up-regulated antitumor cytotoxicity of LAK cells in patients with bladder cancer. The growth of LAK cells induced by IL-2 was down-regulated by hydroxyl radical. The effects of these reactive oxygen species on the proliferation of LAK cells induced by IL-2 were different. PMID- 11918853 TI - Effect of astragaloside IV on T, B lymphocyte proliferation and peritoneal macrophage function in mice. AB - AIM: To investigate the effect of astragaloside IV (ASI) on T, B lymphocyte proliferation, antibody production, and cytokines produced by murine peritoneal macrophages. METHODS: MTT assay was used to determine T, B lymphocyte proliferation and quantitative hemolysin spectrophotometry (QHS) assay was applied to test antibody production; IL-1 production was measured by thymocyte proliferation assay; TNF-alpha production was determined by the cytotoxicity assay against L929 cells. RESULTS: 1) In vivo, ASI 50-200 mg/kg ig for 7 d increased T lymphocyte proliferation and antibody production, and ASI 50-100 mg/kg ig for 7 d increased B lymphocyte proliferation but ASI 200 mg/kg had no effect on B lymphocyte proliferation; 2) In vitro, ASI increased T, B lymphocyte proliferation only at 100 nmol/L; 3) ASI increased IL-1 activity at 1 nmol/L in vitro, but decreased it at 100 and 1000 nmol/L; 4) ASI inhibited TNF-alpha activity with or without LPS-stimulation in vitro. CONCLUSION: ASI increased T, B lymphocyte proliferation and antibody production in vivo and in vitro; but inhibited productions of IL-1 and TNF-alpha from peritoneal macrophages in vitro. PMID- 11918855 TI - Inhibition of beta-estradiol on trachea smooth muscle contraction in vitro and in vivo. AB - AIM: To investigate the effect of beta-estradiol on trachea smooth muscle contraction in vitro and in vivo. METHODS: (1) Rabbit tracheas were incubated in organ baths filled with Krebs solution and supplied with a mixed gas of 95 % O2 and 5 % CO2. The isometric force was measured by ink-writing recorders. (2) The incubation period of asthma induced by histamine and acetylcholine (ACh) in guinea pig were measured before and after beta-estradiol (1 mg/kg) were given intramuscularly. RESULTS: (1) Administration of beta-estradiol (0.1 mmol/L) caused relaxation of isolated trachea muscle strips (TMS) in rabbits pre contracted by ACh and KCl (39 % +/- 5 % and 45 % +/- 19 %). The presence of indomethacin or methylene blue partly decreased the relaxation to beta-estradiol (26 % +/- 8 % and 28 % +/- 13 %), but Nomega-nitro-L-arginine (L-NNA) and propranolol and epithelium removal did not affect it (38 % +/- 10 %, 40 % +/- 15 %, 37 % +/- 8 %). beta-Estradiol can shifted the concentration-response curves of ACh and CaCl2 to the rightward (pD2 = 3.98 and 4.75). In addition, it could also significantly inhibit the contraction of phase caused by ACh, but did not affect the contraction of phase II caused by CaCl2. (2) The incubation period of asthma in guinea pig were delayed by beta-estradiol (1 mg/kg) given intramuscularly. CONCLUSION: (1) The relaxation of beta-estradiol in vitro was epithelium independent and associated with the inhibition of potential-dependent channel and release of Ca2+ from sarcoplasm reticulum induced by ACh. In addition, release of prostaglandins from trachea smooth muscle cells and relaxation through cGMP approach were also included. beta-Adrenoceptor-mediated relaxation was not involved. (2) beta-Estradiol can relax the trachea in vivo in guniea pig. PMID- 11918854 TI - Effects of theophylline on CD4+ T lymphocyte, interleukin-5, and interferon gamma in induced sputum of asthmatic subjects. AB - AIM: To investigate the mechanism of anti-inflammatory action of theophylline on asthma. METHODS: Nineteen asthmatic subjects were administered 200 mg sustained release theophylline twice daily for 4 weeks. The number of CD4+ T lymphocytes, eosinophils, and the levels of interleukin (IL)-5 and interferon gamma (IFN gamma) in sputum pre- and post-administration were measured by direct immunofluorescene technique, Wright's stain and enzyme-linked immunosorbent assay, respectively. The symptom scores and lung function were also evaluated. RESULTS: Theophylline treatment significantly improved symptom scores and FEV1.0, FEV1.0 % (P < 0.05) and reduced sputum eosinophils (P < 0.01) in asthmatic subjects. These were accompanied by a decrease in sputum IL-5 level (P < 0.01), but sputum CD4+ T lymphocytes and IFN-gamma had no significant change (P >0.05). The mean (range) serum theophylline concentration in final steady state was 7.9 (3.9 - 12.9) mg/L. CONCLUSION: The anti-inflammatory action of theophylline in asthma may result from reduction of IL-5 production in the airways. PMID- 11918856 TI - Pharmacodynamics and pharmacokinetics of inhaled nitric oxide in dogs with septic acute respiratory distress syndrome. AB - AIM: To evaluate pharmacodynamics and pharmacokinetics of inhaled nitric oxide (iNO) in dogs with acute respiratory distress syndrome (ARDS). METHODS: ARDS, induced after iv injection of endotoxin, was evidenced by reduction of paO2/FiO2 from (62.5 +/- 2.8) to (26 +/- 4) kPa and dynamic lung compliance (Cdyn) from (14.8 +/- 0.7) to (8.6 +/- 0.6) mL.kPa-1 . kg-1, increase of dead space (VD/VT) from (0.14 +/- 0.06) to (0.58 +/- 0.05), intrapulmonary shunting (Qs/Qt) from 4.7 % +/- 1.7 % to 39 % +/- 7 %, and pulmonary vascular resistance index (PVRI) from (16 +/- 4) to (51 +/- 8) kPa.s.L-1 . m-2 (all P < 0.05), along with severe intrapulmonary neutrophil recruitment and peripheral neutropenia. The animals were then treated as either a control or an NO group (n = 6 each, iNO 0.4 - 3.2 micromol/L) for another 10 h. RESULTS: More survival was found in NO group (4/6 vs 0/6, P < 0.05). iNO at 0.8, 1.6, and 3.2 micromol/L (20, 40, and 80 ppm) resulted in > 40 % increase of paO2/FiO2 and Cdyn, a reduction of VD/VT to 0.32, Qs/Qt to < 25 %, and PVRI by > 50 % (30.8 kPa . s . L-1 . m-2) compared to the control. Optimal iNO dose was around 0.8 micromol/L as higher methemoglobin (MetHb, > 3 %) was found at higher NO. iNO had no adverse effects on surfactant phospholipids and lung fluid balance, but attenuated expression of tumor necrosis factor alpha,beta2 integrin CD11b, and interleukin-8 mRNA in the lungs by 22 %, 44 %, and 25 %, respectively (P < 0.05). CONCLUSION: Pharmacodynamics of iNO in this model was related to improvement in gas exchange, Cdyn, PVRI, and suppression of proinflammatory cytokine expression in the lungs, and its adverse effect was mainly confined to MetHb at higher NO dose. PMID- 11918857 TI - Determination of domperidone in human plasma by LC-MS and its pharmacokinetics in healthy Chinese volunteers. AB - AIM: To determine the concentration of domperidone in human plasma and investigate its pharmacokinetics in Chinese male volunteers following oral administration of a single dose of the medicine 20 mg. METHODS: Plasma samples were processed by liquid-liquid extraction and a liquid chromatographic-mass spectrometric assay was developed for the determination of domperidone in human plasma. RESULTS: Assay linearity was obtained in the range of 0.52 - 154.5 microg/L (r = 0.9999). The recovery of domperidone from human plasma was more than 75 %. The intraday and interday relative standard deviations (RSD) for the lowest concentration examined (0.52 microg/L) were 9.4 % and 7.6 %, respectively. The method was utilized to determine the concentration of domperidone in Chinese male volunteers. The concentration-time curve was fitted to a two-compartment model. Its main pharmacokinetic parameters were: Tmax = (0.8 +/- 0.7) h, Cmax = (50 +/- 32) microg/L, T1/2 = (7.8 +/- 1.6) h. CONCLUSION: The method described in this report was of high sensitivity and selectivity for accurate determination of the plasma concentration of domperidone in human. PMID- 11918858 TI - How now, brown cow, vaccine or extract? PMID- 11918859 TI - Inhibition of phosphodiesterase 4 in allergic rhinitis. PMID- 11918860 TI - Human nasal allergen provocation for determination of true allergic rhinitis: methods for clinicians. AB - The nasal provocation test (NPT) could be more extensively used in the diagnosis of allergic rhinitis by practicing physicians. However, the procedure has not been standardized, and has mainly been utilized for scientific purposes in the US. This review illustrates a wide variety of techniques and approaches to dosing and concentration of allergen extracts, and delivery systems. It also outlines the lack of a unified outcomes-evaluation system, including clinical symptom scores and nasal patency measurements, in different countries. NPT is a safe, simple, and useful method when conducted with the consideration of indications and contraindications. Standardized NPT has the potential to become a more frequently used additional clinical test in the diagnosis of allergic rhinitis. PMID- 11918861 TI - Thoughts on the pathophysiology of nonallergic rhinitis. AB - Nonallergic noninfectious rhinitis is a diagnosis by exclusion, meaning that a number of poorly defined nasal conditions that have in common allergy and infection as a cause of the rhinitis have been excluded. The etiology of some subgroups of nonallergic noninfectious rhinitis, like nonallergic rhinitis with eosinophilia (NARES) and drug-induced rhinitis, are quite well defined, but in the majority of the patients, the etiology and pathophysiology are unknown. These patients are classified as idiopathic rhinitis patients. A careful determination of the intensity of the symptoms combined with modern diagnostic tools enables us to discriminate idiopathic rhinitis patients from normal controls. This review discusses the possible pathophysicologic mechanisms of nonallergic noninfectious rhinitis, with emphasis on idiopathic rhinitis. PMID- 11918862 TI - Neurology of allergic inflammation and rhinitis. AB - Afferent nerves, derived from the trigeminal ganglion, and postganglionic autonomic nerves, derived from sympathetic and parasympathetic ganglia expressing many different neurotransmitters, innervate the nose. Reflexes that serve to optimize the air-conditioning function of the nose by altering sinus blood flow, or serve to protect the nasal mucosal surface by mucus secretion, vasodilatation, and sneezing, can be initiated by a variety of stimuli, including allergen, cold air, and chemical irritation. Activation of nasal afferent nerves can also have profound effects on respiration, heart rate, blood pressure, and airway caliber (the diving response). Dysregulation of the nerves in the nose plays an integral role in the pathogenesis of allergic rhinitis. Axon reflexes can precipitate inflammatory responses in the nose, resulting in plasma extravasation and inflammatory cell recruitment, while allergic inflammation can produce neuronal hyper-responsiveness. Targeting the neuronal dysregulation in the nose may be beneficial in treating upper airway disease. PMID- 11918863 TI - Sedation, cognition, and antihistamines. AB - First-generation antihistamines are well-known to cause subjective drowsiness. A myriad of studies has also been published that suggest a clear relationship between the use of these drugs and objective performance impairment. Although not all of the tests used in these studies have been validated, the data are fairly consistent, and suggest a difference between earlier (first-generation) sedating antihistamines and the newer (second-generation) nonsedating antihistamines. PMID- 11918864 TI - The cost of treating allergic rhinitis. AB - Allergic rhinitis is a high-cost, high-prevalence disease. In the year 2000, over $6 billion was spent on prescription medications to treat this illness. Although it is not associated with severe morbidity and mortality, allergic rhinitis has a major effect on the quality of life of the more than 50 million Americans with this illness. Intranasal corticosteroids (INCS) and nonsedating antihistamines (NSAH) are the most common prescription medications for this disease. INCS are recognized as the most effective treatment regimen for chronic symptoms. NSAH are perceived as important in the treatment of patients with mild disease, or as add on therapy to INCS. When the literature is reviewed, the INCS produce the greatest decrease in total nasal symptom scores, the largest effect size, when compared with NSAH. Both classes of medications produce similar effects on concurrent allergic conjunctivitis. Further recent studies indicate that the INCS are also superior when used on an as-needed basis, and that there is little clinical benefit from the addition of loratadine to intranasal fluticasone. INCS have lower average wholesale prices as a class than the NSAH. Since the INCS are the dominant medication in efficacy studies and cost less, cost-effectiveness studies always favor intranasal corticosteroids. PMID- 11918865 TI - Local IgE synthesis in allergic rhinitis and asthma. AB - Local IgE production has been a contentious concept for over 40 years. Although a host of clinical observations suggested a local source of IgE, co-localization of IgE to B cells in the nasal mucosa could not be demonstrated. In contrast, igE producing B cells were observed in local lymphoid tissue. Definitive proof of local IgE synthesis in the nasal mucosa has now become available. Here, we review the recent developments in local IgE research in allergic rhinitis and asthma within their historical context, and speculate on the implications of both local IgE synthesis and ongoing research into local immunoglobulin isotype switching for patient therapy. PMID- 11918867 TI - Nasal congestion and airway obstruction: the validity of available objective and subjective measures. AB - Rhinomanometry and acoustic rhinometry provide the best methods for objective assessment of nasal obstruction. Advanced equipment for these methods is now available, and most devices are reliable provided that care is taken to calibrate the device properly and, for rhinomanometers, the user is completely familiar with the mathematical algorithm for resistance used by the accompanying software. Suggestions for improvement in standardization of rhinomanometry are given. Rhinomanometry and rhinometry are both capable of objectively measuring nasal obstruction, but they are complementary methods that assess different nasal attributes, the former being a test of nasal function, and the latter a representation of nasal geometry. Objective methods are strongly recommended for use in the evaluation of pharmacologic agents that are expected to improve nasal airflow. When further studies needed to validate their use for long-term comparisons are done, these methods should find an increasing place in clinical practice. PMID- 11918866 TI - The role of leukotriene inhibitors in allergic rhinitis and paranasal sinusitis. AB - Leukotrienes are inflammatory mediators that are known as the slow-reacting substance of anaphylaxis produced by a number of cell types, including mast cells, eosinophils, basophils, macrophages, and monocytes. Synthesis of these meditators in the text result from the cleavage of arachidonic acid in cell membranes, and exert their biologic effects by binding and activating specific adaptors. This occurs in a serious of events that lead to contraction of the human airway smooth muscle, chemotaxis, and increased vascular permeability. These effects have lead to their important role in the diseases of asthma, allergic rhinitis, and possible paranasal sinusitis with the formation of nasal polyps. Since these agents lead to the production of symptoms in patients who are asthmatic, the use of leukotriene enzyme inhibitors, particularly montelukast, zafirlukasts seem appropriate. PMID- 11918868 TI - Staphylococcus aureus superantigens and airway disease. AB - Twenty-five percent of the population are permanent carriers of Staphylococcus aureus, possibly producing a variety of toxins with superantigenic properties. Staphylococcal superantigens are a group of high- molecular-weight pyrogenic proteins that have in common an extremely potent stimulatory activity for T lymphocytes, macrophages, mast cells, eosinophils, and epithelial cells. The role of staphylococcal superantigens in atopic dermatitis has recently been recognized, and new evidence suggests that similar mechanisms may also be relevant in airway disease. This circumstantial evidence is currently limited to rhinitis, sinusitis, and possibly asthma, but may, if supported, open a new understanding of pathomechanisms and therapeutic targets. PMID- 11918870 TI - Blending art and science in healthcare, or the progressive blurring of traditional specialty boundaries. PMID- 11918869 TI - TGF-beta-mediated control of allergen-specific T-cell responses. AB - Control of allergen-specific response by suppressive cytokines involves several layers of regulation, including secretion of the cytokine, deviation of cytokine expression by altered T-cell differentiation, immediate (de-) phosphorylation events upon binding of suppressive cytokines, and laterations in susceptibility of suppression. PMID- 11918871 TI - An assessment of surgical education. AB - A review of the dramatic changes in society, science and medicine that have affected the time we have available for education of students and residents. Reference is made to distance learning, educational efficiency and mental practice as concepts that may aid educators in the quest to provide the public with well trained surgeons. Surgical educators are urged to look outside of traditional models of teaching and evaluating for tools that have been successfully used by industry or business. PMID- 11918872 TI - Quality of life and symptomatic outcome three to five years after laparoscopic Toupet fundoplication in gastroesophageal reflux disease patients with impaired esophageal motility. AB - BACKGROUND: Many centers practice a tailored approach to laparoscopic antireflux surgery in attempt to prevent postoperative side effects in gastroesophageal reflux disease (GERD) patients with an impaired esophageal motility. As a result of controversial findings reported in literature no worldwide accepted consensus exists regarding the appropriate indication for this tailored approach. The aim of this prospective study was to evaluate quality of life and symptomatic outcome in selected patients for a follow-up of 3 to 5 years. METHODS: A total of 155 patients with esophageal dismotility underwent laparoscopic Toupet fundoplication (LTF). Basic requirements for surgery included in all patients a detailed evaluation of symptoms and quality of life (Gastrointestinal Quality of Life Index [GIQLI]), esophagogastroduodenoscopy, 24-hour pH monitoring, and esophageal manometry. Patients were evaluated 6 weeks, 3 months, 1 year, and 3 to 5 years after LTF. RESULTS: GERD-related symptoms such as heartburn, regurgitation, dysphagia, or chest pain showed a significant improvement (P <0.05 to 0.001) in all gradings immediately after surgery. During the complete follow-up, a total of 4 patients (2.6%) required laparoscopic redo surgery because of recurrent GERD symptoms. Two patients (1.3%) were adequately maintained on short-term proton pump inhibitor therapy because of mild symptoms. All these patients have shown a pathological DeMeester score within the early period after surgery (3 months or 1 year control). Severe and persistent side effects have been present in 7 patients (4.5%), mild to moderate side effects in 11 patients (7.1%). Other side effects have been temporary and resolved spontaneously. GIQLI improved significantly (P <0.05 to 0.01) in all dimensions and persisted for at least 5 years with mean values comparable with healthy individuals. CONCLUSIONS: LTF is effective, well tolerated, and improves quality of life, improving long-term outcome with an acceptable rate of long-term side effects in GERD patients with moderate to severe esophageal dismotility for a follow-up period of 3 to 5 years. PMID- 11918873 TI - An assessment of vertical banded gastroplasty-Roux-en-Y gastric bypass for the treatment of morbid obesity. AB - OBJECTIVE: To analyze retrospectively the mortality, morbidity, and weight loss of a specific form of gastric bypass for the treatment of morbid obesity. The technique incorporates a small pouch along the lesser curvature of the stomach, an outlet restricted by a nondistensible band and a Roux-en-Y gastric bypass. MATERIAL AND METHODS: We analyzed 652 consecutive patients with no previous bariatric surgery who underwent our present form of gastric bypass. Parameters used to evaluate the technique included mortality, weight loss at 5 years and complications. The operation is a combination of vertical banded gastroplasty and Roux-en-Y gastric bypass (VBG-RGB). The patients followed up to 5 years had an initial weight of 140 kg [range, 94 to 288] and a BMI of 50 [range, 38 to 86]. Superobese individuals (BMI of 60 [range, 48 to 86]) made up 42% of the group. RESULTS: There was an early reoperation rate of 0.5%. The incidence of late complications that required reoperation was 0.5%. There were 2 deaths in the study from pulmonary embolism for a mortality of 0.3%. At 5 years, the patients had lost an average of 58kg [range, 14 to 143] and had a percentage excess weight loss of 77 [range, 32 to 108]. Their BMI was reduced to 29 kg/m(2) [range, 20 to 43] and 93% lost more than 50% of the excess weight. CONCLUSIONS: VBG-RGB is effective in producing superior weight loss in morbid and superobese patients and has a low mortality and morbidity. We recommend this procedure without reservations. PMID- 11918874 TI - Detection of colorectal lesions with virtual computed tomographic colonography. AB - BACKGROUND: The aim of our study was to compare the performance of virtual computed tomographic colonography with that of conventional colonoscopy in a blinded, prospective study in 165 patients with suspected colorectal lesions. METHODS: There were 165 patients, all referred for conventional colonoscopy, who underwent preliminary virtual computed tomographic colonography. Computed tomograhic images of all suspected lesions were analyzed and subsequently compared with conventional colonoscopy findings. RESULTS: There were 30 colorectal cancers and 37 polyps identified at conventional colonoscopy. Virtual computed tomographic colonography correctly detected all cancers, as well as 11 of 12 polyps of 10 mm in diameter or larger (sensitivity, 92%); 14 of 17 polyps between 6 and 9 mm (sensitivity, 82%); and 4 of 8 polyps of 5 mm or smaller (sensitivity, 50%). The per-patient sensitivity and specificity were 92% and 97%, respectively. CONCLUSIONS: Virtual computed tomographic colonography has a diagnostic sensitivity similar to that of conventional colonoscopy for the detection of colorectal lesions larger than 6 mm in diameter. PMID- 11918875 TI - Validation of venous leg ulcer guidelines in the United States and United Kingdom. AB - BACKGROUND: Venous leg ulcers account for 85% of all lower-extremity ulcers, with treatment costs of 3 billion dollars and loss of 2 million workdays per year. The purpose of this study was to validate the clinical efficacy and cost effectiveness of multidisciplinary guidelines for the diagnosis and treatment of venous leg ulcers. METHODS: Eighty (40 retrospective, 40 prospective) patients from the United States and United Kingdom were enrolled. RESULTS: United States patients were 6.5 times and United Kingdom 2 times more likely to heal if a guideline was followed (P <0.001). A significant decrease was noted in healing time for both the United States and United Kingdom (P <0.01), and the median cost decreased significantly when the guideline was followed (P <0.01). CONCLUSIONS: Implementation of a guideline for diagnosis and treatment of venous leg ulcers resulted in improvement in diagnosis, decrease in healing time, and an increase in healing rates resulting in lower costs. PMID- 11918876 TI - Pancreaticojejunostomy with invagination of spatulated pancreatic stump into a jejunal pouch. AB - BACKGROUND: Pancreaticoenteric anastomosis after pancreatic resection is of major concern as anastomotic leak continues to be common. There is no unanimity for the preferred technique and overall incidence of pancreatic leak is reported to be 2% to 14%. METHODS: A new safe method of anastomosing pancreatic stump to a jejunal pouch is described. A 15-cm length of the jejunal end is detubularized and reconfigurated into a U-shaped patch. The pancreatic stump is mobilized for about 3 cm and the duct is spatulated posteriorly and anastomosed to a cut in convex margins of the patch. This is converted into a pouch invaginating the spatulated pancreaticojejunal anastomosis. RESULTS: Eleven cases of periampullary malignancy after pancreaticoduodenectomy have been operated on with no pancreatic leak. CONCLUSIONS: Even a bulky pancreas can be invaginated into the pouch. The resulting anastomosis is completely intraluminal. Spatulated mucosa to mucosa anastomosis should reduce the chances of late stenosis. PMID- 11918877 TI - Is attrition of advanced trauma life support acquired skills affected by trauma patient volume? AB - BACKGROUND: We assessed the effect of trauma volume on skills attrition among physicians completing the advance trauma life support (ATLS) course. METHODS: Cognitive (40 item multiple choice question [MCQ] examination) and clinical (4 objective structured clinical examinations [OSCE] trauma stations) performances were compared among physicians who completed the ATLS course, subdividing them into groups treating more than 50 and fewer than 50 trauma patients per year. Both groups had 12 physicians from six periods (n = 144) related to time of course completion: immediate (0), 6 months, 2 years, 4 years, 6 years, and 8 years after ATLS. OSCE scores (maximum standardized: 20), the degree of adherence to priorities (priority score: range 1 to 7), the degree of organized approach (approach score: range from 1 to 5) were compared. RESULTS: The mean precourse MCQ scores (59.4% to 62.4%) were similar for both groups. Immediate and progressive cognitive skill attrition and detailed clinical skill attrition were worse in the low volume group. Global skills (organized approach and adherence to priorities) were preserved similarly for at least 8 years in all groups. CONCLUSIONS: Our data suggest that trauma volume affects trauma skills attrition. PMID- 11918879 TI - The outcome of early pressure-controlled inverse ratio ventilation on patients with severe acute respiratory distress syndrome in surgical intensive care unit. AB - BACKGROUND: Pressure-controlled inverse ratio ventilation (PC-IRV) was used in patients with acute respiratory distress syndrome (ARDS) after failed volume cycled conventional ratio ventilation (VC-CRV). The aim of this study was to evaluate the outcome of early PC-IRV in severe ARDS. METHODS: Twenty patients with severe ARDS were switched from VC-CRV to PC-IRV if they failed to maintain SaO(2) >90% by the following criteria: peak inspiratory pressure (PIP) >35 cm H(2)O, FIO(2) = 60%, and positive end-expiratory pressure (PEEP) 10 cm H(2)O. RESULTS: The values of PIP, mean airway pressure, minute volumes, and lung injury score in VC-CRV were 43.9 +/- 8.0 cm H(2)O, 19.5 +/- 6.4 cm H(2)O, 11.0 +/- 2.1 L/min, and 2.8 +/- 0.2 respectively. In PC-IRV, the corresponding data were 31.8 +/- 5.1 cm H(2)O, 25.4 +/- 4.6 cm H(2)O, 8.3 +/- 0.9 L/min, and 2.5 +/- 0.4. All of these parameters were significantly different. Fifteen patients (75%) survived their intensive care unit stay. CONCLUSIONS: Early PC-IRV in severe ARDS improves oxygenation, facilitates tapering of high fraction of inspiratory oxygen, and decreases high PEEP or PIP, and then results in the improvement of the patient's outcome. PMID- 11918878 TI - Perceptions of women medical students and their influence on career choice. AB - BACKGROUND: Although women make up nearly half of medical school classes in the United States, just over 20% of residents in surgery are women (excluding obstetrics/gynecology). The objective of this study was to identify whether the proportion of women surgeons on the faculty who have frequent encounters with medical students during their surgery rotation influences the student's perceptions about women surgeons or their career choice. METHODS: Seven US medical schools with proportions of women surgeons on the fulltime faculty varying from 10% to 40% were selected to participate in this survey. Women medical students graduating in the spring of 2000 were asked to complete an anonymous 29 question survey designed to assess their perceptions of women surgeons' career satisfaction. Demographic information about the students such as career choice, age, and marital status was also collected. The differences in responses between those schools with 40% women faculty and those with less than 15% were analyzed. RESULTS: The overall response rate was 74% (305 of 413). Forty five percent of students had daily or weekly contact with a woman surgery attending. There were no differences in perceptions of women surgeons' career satisfaction for those students at schools with 40% women surgeons versus those with less than 15%. However, 21 of 24 (88%) students choosing surgery as a career were from the three schools with a greater number of women surgical role models (P <0.0001). Students who chose a career in surgery perceived the women faculty's career satisfaction to be higher than did those students not choosing a surgical career (P <0.01). CONCLUSIONS: Women medical students perception of women surgeons' career satisfaction did not appear to be affected by the proportion of women surgeons on the faculty at their medical school. However, their choice of surgery as a career was strongly associated with a higher proportion of women on the surgical faculty. PMID- 11918880 TI - Extracorporeal suture technique for suprahepatic caval anastomosis in liver transplantation. AB - BACKGROUND: During liver transplantation, an oversized graft or active bleeding in the hepatic area can make performance of the suprahepatic caval anastomosis extremely difficult or even impossible. In other instances, a brief as possible warm ischemia is desired to provide a marginal graft with maximum chances of good functioning. METHODS: In order to manage those conditions, a suture technique was devised that allows the construction of a substantial part of the suprahepatic caval anastomosis keeping the graft outside the abdomen of the recipient. RESULTS: Over a 12-month period, the technique was applied in 7 out of 148 transplants (5%). The 7 cases presented one or more of the following conditions: significant oversize mismatch (n = 6), active bleeding in the hepatic area (n = 1), and marginal graft (n = 4). Warm ischemia time averaged 27 minutes, a value not significantly different from the mean warm ischemia time of 25 minutes recorded in the easier transplants in which the conventional technique was used (P = 0.2467). CONCLUSIONS: This extracorporeal suture technique allows construction of the suprahepatic caval anastomosis in critical situations arising during liver transplantation and avoidance of the prolonged warm ischemia that could be expected in such cases. PMID- 11918881 TI - Surgery for inoperable breast cancer. AB - Neoadjuvant chemotherapy has the potential to convert inoperable breast cancer into operable disease; however, patients may remain inoperable by the classic criteria after neoadjuvant chemotherapy. In such cases, palliative surgical therapy to promote comfort and hygiene and to control wound breakdown may need to be considered. This report documents this clinical scenario in a patient with a large exophytic breast cancer who had a partial response with neoadjuvant chemotherapy and required an extended radical mastectomy with extensive reconstruction for coverage. The decision to undertake such a surgical procedure is complicated when the patient's life expectancy may be extremely limited and both patient and treatment team must carefully weigh the risks and potential benefits of a highly complex but technically feasible operation. PMID- 11918882 TI - A decade of experience with transthoracic and transhiatal esophagectomy. AB - BACKGROUND: Morbidity and mortality remain significant for transthoracic (TT) and transhiatal (TH) esophagectomy. We report a case-specific approach employing either resection to minimize perioperative morbidity and mortality. METHODS: All primary esophageal resections performed for benign and malignant esophageal disease were reviewed over a 10-year period. The operative approach was tailored to the location and extent of disease and the physiologic reserve of the patient. RESULTS: In all, 115 patients underwent esophagectomy for benign (25) and malignant (90) disease. Fifty-six TT and 59 TH resections were performed. Four emergent TT cases did not have reconstruction. There was 1 hospital mortality. Perioperative transfusion was avoided in 65 patients. Respiratory complications occurred in 15. Three patients had a cervical anastomotic leak requiring open wound drainage. No association between resection type and complication was evident. CONCLUSIONS: The judicious use of both TT and TH esophagectomy resulted in an operative mortality of less than 1%, reduced operative blood loss, and a relatively low rate of perioperative complications. PMID- 11918883 TI - Risk factors predictive of lymph node metastasis in depressed early gastric cancer. AB - BACKGROUND: This study was conducted to identify risk factors predictive of regional lymph node metastasis in depressed early gastric cancer and further to establish an objective criterion useful to indicate additional surgical treatment in cases in which submucosal tumor extension becomes evident by endoscopic mucosal resection (EMR). METHODS: Data from 276 patients surgically treated for depressed early gastric cancer were collected, and the relationship between the patient and tumor characteristics, and the lymph node metastasis was retrospectively evaluated by multivariate analysis. RESULTS: In the multivariate logistic regression model, female sex, a larger tumor size (20 mm or more), submucosal invasion, and presence of lymphatic vessel involvement were found to be independent risk factors for lymph node metastasis. Among 145 patients with submucosally invasive carcinoma, no lymph node metastasis was observed in patients who showed none of the other three risk factors, whereas 14.3% and 23.3% of patients with one and two of these factors had lymph node metastasis, respectively. The lymph node metastasis rate was calculated to be 86.7% in patients who had all three factors. CONCLUSIONS: Submucosal invasion, female sex, tumor size of 20 mm or more, and lymphatic vessel involvement were significantly and independently related to the presence of lymph node metastasis in depressed early gastric cancer. The positive number of the latter three risk factors is a simple criterion to indicate additional surgical treatment in cases with submucosal invasion revealed first by EMR. PMID- 11918884 TI - Total versus selective hepatic vascular exclusion in major liver resections. AB - BACKGROUND: Total hepatic vascular exclusion (THVE) and selective hepatic vascular exclusion (SHVE) are two effective techniques for bleeding control in major hepatic resections. Outcomes of the two procedures were compared. METHODS: Patients undergoing major liver resection were randomly allocated to the THVE and SHVE groups. Intraoperative hemodynamic changes and the postoperative course of the two groups were compared. RESULTS: During vascular clamping, the THVE group showed a significant elevation in pulmonary vascular resistance, systemic vascular resistance, intrapulmonary shunts, and a significant reduction in cardiac index, compared with the SHVE group (P <0.05). Patients undergoing THVE received more crystalloids and blood, showed more severe liver, renal and pancreatic dysfunction, and had a longer hospital stay than the SHVE group (P <0.05). CONCLUSIONS: Both techniques are equally effective in bleeding control in major liver resections. THVE is associated with cardiorespiratory and hemodynamic alterations and may be not tolerated by some patients. SHVE is well tolerated with fewer postoperative complications and shorter hospitalization time. PMID- 11918886 TI - Traumatic transection of the pancreas. PMID- 11918885 TI - Factors predisposing to bacterial invasion and infection. AB - BACKGROUND: Bacterial infections remain important causes of morbidity and mortality in surgical patients. Our understanding of the effects of bacteria on the host, and also the defense mechanisms available to the host, is improving all the time. Modern tools in biochemistry, immunology, and molecular biology have provided powerful methods to further our understanding of the complex interactions that contribute to our host defense response. This review reflects current thinking regarding the factors that contribute to bacterial infection and host defense response. DATA SOURCES: This review was compiled after an extensive review of the current and historical literature, and highlights a number of areas involved in the pathogenesis of bacterial infection. CONCLUSIONS: Bacteria can have a wide-ranging and deleterious effect on the host. Many different therapeutic approaches have been attempted to modulate the host response and limit the deleterious effects of bacteria. As our understanding of the underlying processes improves, these therapies should improve accordingly. PMID- 11918887 TI - Laparoscopic splenectomy for massive splenomegaly. AB - BACKGROUND: Laparoscopic splenectomy (LS) is the preferred operative approach for diseases involving normal-sized spleens. Our experience with laparoscopic splenectomy in the setting of massive splenomegaly is presented. METHODS: A prospective review of patients undergoing LS for massive splenomegaly was conducted. Massive splenomegaly (MS) in adults was defined as a craniocaudal length >or=17 cm or a morcellated weight >or=600 g. In children, spleens measuring fourfold larger than normal for age were considered massive. RESULTS: Forty-nine patients with MS were treated with LS. The most common primary diagnoses were lymphoma and leukemia. Mean splenic length was 20 cm (15 to 27 cm), with weights ranging from 600 to 4,750 g. Twelve patients with supermassive splenomegaly (length >22 cm) required a hand-assisted laparoscopic approach. There were no conversions to open surgery. Mean operating time was 171 minutes (90 to 369). Mean blood loss was 114 cc (<30 to 600 cc). Average length of stay was 2.3 days (1 to 16). Minor postoperative complications occurred in 3 patients. CONCLUSIONS: Laparoscopic splenectomy in the setting of splenomegaly is safe and appears to minimize perioperative morbidity. In patients with supermassive splenomegaly, a hand-assisted laparoscopic approach may be required. PMID- 11918888 TI - Rupture of splenic artery pseudoaneurysm presenting with massive upper gastrointestinal bleed. AB - Splenic artery pseudoaneurysms develop in as many as 10% of patients with acute pancreatitis. The diagnosis should be suspected in patients with pancreatitis who develop either upper gastrointestinal bleeding without an obvious cause or in whom a contrast-enhanced lesion is demonstrated within or adjacent to a suspected pseudocyst as determined by computed tomography scan. PMID- 11918889 TI - Early effects of parathyroidectomy on erythropoietin production in secondary hyperparathyroidism. AB - BACKGROUND: Secondary hyperparathyroidism (2-HPT) has an adverse effect on renal anemia and may cause a hyporesponsiveness to recombinant human erythropoietin (rHuEpo) in patients with chronic renal failure. The early effects of parathyroidectomy (PTx) on renal anemia, erythropoietin production, and nutritional state were examined. METHODS: Twenty-nine patients under hemodialysis therapy received a PTx for 2-HPT. They were prospectively studied regarding hematological parameters, rHuEpo use, plasma erythropoietin levels, and nutritional condition until 12 months after PTx. RESULTS: The hemoglobin level showed a significant increase from 3 months after PTx (10.2% +/- 1.5% to 11.2% +/ 1.3%; P <0.01), associated with a consistent increase of the reticulocyte count. These changes lasted until 12 months after PTx. The plasma erythropoietin level showed a gradual increase of up to about 5 times the level of the preoperative value, until 12 months after PTx (22.6 +/- 10.1 to 106.3 +/- 112.1 mU/mL; P <0.001). The weekly dose of rHuEpo administration decreased after 3 months. The serum levels of albumin and total protein also significantly and gradually improved until 12 months after PTx. CONCLUSIONS: PTx caused a significant early improvement in renal anemia in patients with secondary hyperparathyroidism. This effect may be caused by an enhanced erythropoietin production and may also be partially due to the improved nutritional state after PTx. PMID- 11918890 TI - Pyogenic hepatic abscess with biliary communication. AB - BACKGROUND: The aims of this study were to compare the effectiveness of percutaneous drainage between pyogenic hepatic abscesses with and those without biliary communication and to evaluate the role of endoscopic biliary stent placement in the treatment of communicating abscesses. METHODS: Sixty-one patients with hepatic abscesses underwent percutaneous drainage. For communicating abscesses refractory to drainage alone, endoscopic biliary stenting was performed. RESULTS: After drainage, all patients showed improvement in their clinical condition and reduction in abscess size. Drainage alone completely cured abscesses in none of 10 patients with biliary communication and obstruction, in 3 of 10 with communication but without obstruction, and in 37 of 41 without communication. Endoscopic stenting was completely effective in all 7 patients with persistent discharge from a communicating abscess without biliary obstruction. CONCLUSIONS: Percutaneous drainage is less effective for communicating abscesses without biliary obstruction than for noncommunicating abscesses. Endoscopic biliary stenting is recommended, if drainage does not achieve a cure. PMID- 11918891 TI - Excision and primary closure of pilonidal sinus using a drain for antiseptic wound flushing. AB - BACKGROUND: In the case of pilonidus sinus treated with primary intention surgery the uneventful healing is still difficult to obtain, as indirectly proven by the number of different procedures that have been suggested, such as cyst excision with or without primary closure, excision followed by marsupialisation, and excision followed by skin flap transposition. The procedure described here involves excision and primary closure, with a drain being used to flush the operative cavity with an antiseptic solution. METHODS: Two hundred and forty three patients (173 men and 70 women) were treated by excising the pilonidal sinus and placing a 12F suction drain at the base of the wound, with its tip being brought out in the left gluteal region at least 5 cm laterally to the lower end of the suture. Suction was stopped on the first postoperative day and the drain was cut just above the skin. On day 2, a 5F catheter was inserted through the drain and the cavity was flushed with an antiseptic solution followed by sterile saline solution; the same treatment was repeated on days 4 and 6. The drain was removed on day 8 or 9, some of the stitches on day 8 or 9 and the rest on day 9 or 10. The surgery was performed on a day hospital basis in 207 cases; the remaining 36 were hospitalized overnight and discharged on the following day. RESULTS: Healing was always by first intention, with none of the 243 patients experiencing any complications. The postoperative follow-up now ranges from 5 to 15 years, and there have not been any recurrences. CONCLUSIONS: The drainage of blood from the bottom of the wound and the use of antiseptic/saline flushing are essential for primary intention healing and the avoidance of recurrences. PMID- 11918892 TI - Neoplastic masquerade syndromes. AB - Masquerade syndromes are classically defined as entities which emulate inflammatory conditions but which are in fact due to a neoplastic process. Careful history and examination in concert with appropriate ancillary investigations and histopathologic evaluation of tissue specimens are required in order to make the correct diagnosis. Many conditions may result in an appearance mimicking an inflammatory condition. The authors review neoplastic conditions which may be considered masquerades. The most common of these is primary intraocular lymphoma or primary central nervous system lymphoma, occurring predominately in older individuals. Diagnostic strategies, therapy, and prognosis are reviewed in detail. Other conditions that can be considered masquerade syndromes are reviewed as well, including lymphomatous and nonlymphomatous conditions, such as melanoma, retinoblastoma, juvenile xanthogranuloma, metastatic lesions, and paraneoplastic syndromes, among others. PMID- 11918893 TI - Retinal pathology occurring after excimer laser surgery or phakic intraocular lens implantation: evaluation of possible relationship. AB - The increasing number of patients undergoing refractive surgery has led to an awareness of the potential retinal complications of these procedures. The purpose of this review is to summarize the reports of retinal pathology and myopic maculopathy that have occurred after excimer refractive surgery or implantation of phakic intraocular lenses, and to evaluate theoretical pathogenetic mechanisms. We found it reasonable to conclude that retinal detachments and macular hemorrhages are not caused by laser surgery, but are rather characteristic of the natural history in the myopic eye. However, although there is no clear-cut evidence for a cause-and-effect relationship between excimer laser surgery and retinal pathology, it is very important to inform patients that refractive surgery only corrects the refractive aspect of myopia, and that the myopia itself still has the potential for serious complications. PMID- 11918895 TI - The evaluation of isolated third nerve palsy revisited: an update on the evolving role of magnetic resonance, computed tomography, and catheter angiography. AB - The evaluation and management of the neurologically isolated third nerve palsy continues to evolve. The major concern for the clinician confronted with a patient with a third nerve palsy has been the exclusion of an intracranial aneurysm. The evolution of new imaging techniques, such as computed tomography angiography and magnetic resonance angiography, have provided new imaging options for clinicians. This article reviews the pertinent recent literature on the use of these imaging studies in evaluating the patient with a third nerve palsy. PMID- 11918896 TI - Identification of progressive glaucomatous visual field loss. AB - In normal individuals, visual field measures are not perfectly repeatable and individual test locations exhibit both short- and long-term sensitivity variations. This physiologic variability is greatly increased in glaucoma and confounds detection of real progressive loss in visual function. Distinguishing progressive glaucomatous visual field loss from test variability therefore represents a complex task. Procedures used for detection of glaucomatous visual field progression may be broadly grouped into four categories: 1) clinical judgment, which consists of simple subjective observation of sequential visual field test results; 2) defect classification systems, whereby specific criteria are used to stratify field loss by discrete score and define progression as score change over time, such as the Advanced Glaucoma Intervention Study scoring system; 3) trend analyses, which follow test parameters sequentially over time to determine the magnitude and significance of patterns within the data, for example linear regression; and 4) event analyses, which identify single events of significant change relative to a reference examination. All of these methods demonstrate distinct benefits and drawbacks, making each useful in specific circumstances, although no single method appears universally ideal. At the present time the best method of detection of progression may be to rely upon confirmation of change at successive examinations and also by correlation of visual field changes with other clinical observations. Alternative analysis methods may become available in the near future to help identify cases of progressive loss. PMID- 11918897 TI - Collagen corneal shields. AB - Collagen corneal shields were developed as a corneal bandage lens and are currently indicated for ocular surface protection following surgery and in traumatic and nontraumatic corneal conditions. Collagen shields are manufactured from porcine or bovine collagen and three different collagen shields are currently available with dissolution times of 12, 24, and 72 hours. The theoretical, experimental, and clinical evidence supports a role for collagen corneal shields as a drug delivery device and in the promotion of epithelial and stromal healing. Presoaking the collagen shield in a pharmacological agent with adjunctive topical treatment represents the most efficacious method of utilizing collagen shields for drug delivery. In microbial keratitis collagen shields can enhance drug delivery, promote epithelial and stromal healing, neutralize collagenases, and reduce corneal inflammation. This review will examine the evidence that supports the role of collagen shields in drug delivery and corneal wound healing. Despite a large volume of experimental (animal) work, studies on human subjects, particularly randomized controlled trials, are lacking. The authors are advocating the reassessment of the application and benefits of corneal collagen shields to clinical practice. PMID- 11918898 TI - The disk edema dilemma. AB - A 33-year-old diabetic woman experienced visual loss OS and disk edema OU. Extensive evaluation led to the diagnosis of diabetic papillopathy. Appropriate evaluation and management of diabetic papillopathy is discussed. PMID- 11918899 TI - Regina Salomea Pilsztynowa, ophthalmologist in 18th-century Poland. AB - Regina Salomea Rusiecki (later Pilsztynowa) was a young, poorly educated Polish Catholic woman in the 18th century who became well known in Eastern Europe and the Ottoman Empire because of her skill in the treatment of cataracts and some other medical problems. She was born in 1718, and at the age of 14 she married an experienced, but significantly older, German Lutheran physician and ophthalmologist, Jakob Halpir. By helping him eagerly in his work, Pilsztynowa learned a great deal of her husband's way of treating cataracts and other ailments, and she ambitiously made use of the friendship and help of other physicians whom she met on her travels. Although there are some occurrences that could lead one to question aspects of Pilsztynowa's straight and honest character, one has also to point to her readiness to help where she could. PMID- 11918901 TI - A step forward for humanity? PMID- 11918900 TI - Anesthesia for intraocular surgery: new anatomic considerations. PMID- 11918902 TI - The mammography screening debate: time to move on. PMID- 11918903 TI - Dystrophin-associated protein complex and heart failure. PMID- 11918905 TI - Painful hip dislocation in cerebral palsy. PMID- 11918904 TI - Paid non-related living organ donation: Horn of Plenty or Pandora's box? PMID- 11918906 TI - Hot, cold, and now bubble fusion. PMID- 11918907 TI - Long-term effects of mammography screening: updated overview of the Swedish randomised trials. AB - BACKGROUND: There has been much debate about the value of screening mammography. Here we update the overview of the Swedish randomised controlled trials on mammography screening up to and including 1996. The Kopparberg part of the Two County trial was not available for the overview, but the continuation of the Malmo trial (MMST II) has been added. The article also contains basic data from the trials that have not been presented before. Methods The trials (n=247010, invited group 129750, control group 117260) have been followed up by record linkage to the Swedish Cancer and Cause of Death Registers. The relative risks (RR) for breast cancer death and mortality were calculated for the invited and the control groups. The trial-specific as well as the age-specific effects were analysed. RRs were calculated by the density method, with total person-time experience of the cohort by time interval of follow-up as a basis for estimating mortality rates. We calculated weighted RRs and 95% CI with the Mantel-Haenszel procedure. FINDINGS: The median trial time-the time from randomisation until the first round was completed for the control group or if the control group was not invited, until end of follow-up-was 6.5 years (range 3.0-18.1). The median follow up time, the time from randomisation, to the end of follow-up, was 15.8 years (5.8-20.2). There were 511 breast cancer deaths in 1864770 women-years in the invited groups and 584 breast cancer deaths in 1688440 women-years in the control groups, a significant 21% reduction in breast cancer mortality (RR=0.79, 95% CI 0.70-0.89). The reduction was greatest in the age group 60-69 years at entry (33%). Looking at 5-year age groups, there were statistically significant effects in the age groups 55-59, 60-64, and 65-69 years (RR=0.76, 0.68, and 0.69, respectively). There was a small effect in women 50-54 years at randomisation (RR=0.95). The benefit in terms of cumulative breast cancer mortality started to emerge at about 4 years after randomisation and continued to increase to about 10 years. Thereafter the benefit in absolute terms was maintained throughout the period of observation. The age-adjusted relative risk for the total mortality was 0.98 (0.96-1.00). INTERPRETATION: The advantageous effect of breast screening on breast cancer mortality persists after long-term follow-up. The recent criticism against the Swedish randomised controlled trials is misleading and scientifically unfounded. PMID- 11918908 TI - Uses of error. A breathtaking patient. PMID- 11918909 TI - Myocardial salvage after coronary stenting plus abciximab versus fibrinolysis plus abciximab in patients with acute myocardial infarction: a randomised trial. AB - BACKGROUND: Patients with acute myocardial infarction might benefit from the addition of glycoprotein IIb/IIIa inhibitors to fibrinolytic or mechanical reperfusion strategies. We compared two strategies, stenting and fibrinolysis, both combined with abciximab, in terms of their ability to salvage myocardium in patients with acute myocardial infarction. METHODS: We enrolled 162 patients with acute myocardial infarction within 12 h of onset of symptoms, assigning 81 stenting plus abciximab and 81 alteplase plus abciximab. Technetium-99m sestamibi scintigraphy was done at admission and after a median of 11 days to calculate initial perfusion defect, final infarct size, and degree of myocardial salvage. The primary endpoint was the salvage index (the ratio of the degree of myocardial salvage to the initial perfusion defect). Major adverse clinical events within 6 months from randomisation were also compared between the two treatments. FINDINGS: Paired scintigraphic measurements were available for 70 patients in the stent group and 71 in the alteplase group. Stenting was associated with greater myocardial salvage than alteplase (median 13.6% [IQR 5.9-23.9] vs 8.0% [2.5-16.0] of the left ventricle; p=0.007). Salvage index was greater in the stent group than in the alteplase group (median 0.60 [0.37-0.82] vs 0.41 [0.13-0.58]; p=0.001). The 6-month mortality rate was 5% (four deaths) in the stent group and 9% (seven deaths) in the alteplase group (relative risk 0.56 [95% CI 0.17-1.88]; p=0.35). INTERPRETATION: In patients with acute myocardial infarction, a reperfusion strategy based on stenting with abciximab produced more myocardial salvage than the combination of fibrinolysis plus abciximab. Larger studies are needed to assess whether these effects translate into clinical benefit. PMID- 11918910 TI - On the back burner. PMID- 11918911 TI - Relation between use of angiotensin-converting enzyme inhibitors and muscle strength and physical function in older women: an observational study. AB - BACKGROUND: Angiotensin-converting enzyme (ACE) inhibitors prevent decline in physical function in patients with congestive heart failure (CHF). We aimed to see whether ACE inhibitors also prevent reduction in physical performance and in muscle strength in older women who do not have CHF. METHODS: We assessed 3-year rates of decline in both knee extensor muscle strength and walking speed in 641 women with hypertension who had participated in the Women's Health and Aging Study. Women were stratified into four groups according to type and duration of antihypertensive drug treatment. 61 had used ACE inhibitors continuously, 133 intermittently, 146 never, and 301 had used other hypertensive drugs either continuously or intermittently. FINDINGS: Participants who had taken ACE inhibitors continuously had a lower mean 3-year decline in muscle strength of 1.0 kg (SE 1.1) compared with -3.7 (0.5) kg in continuous/intermittent users of other antihypertensive drugs (p=0.016) and with -3.9 kg in those who had never used antihypertensives (p=0.026). Muscle strength fell by 3.0 kg in 3 years in both continuous and intermittent users of ACE inhibitors (p=0.096). Mean 3-year decline in walking speed in continuous ACE inhibitor users was -1.7 cm/s compared with -13.6 cm/s in intermittent users of ACE inhibitors (p=0.015), -15.7 cm/s in continuous/intermittent users of other antihypertensive drugs (p=0.002), and 17.9 cm/s in never users of antihypertensive drugs (p=0.001). INTERPRETATION: ACE inhibitor treatment may halt or slow decline in muscle strength in elderly women with hypertension and without CHF. PMID- 11918912 TI - Age at acquisition of Helicobacter pylori infection: a follow-up study from infancy to adulthood. AB - BACKGROUND: Helicobacter pylori infection is common worldwide, but the time of acquisition is unclear. We investigated this issue in a cohort of children selected retrospectively from a population followed up for 21 years. METHODS: We monitored 224 children (99 black, 125 white; 110 male, 114 female) from 1975-76 (ages 1-3 years) to 1995-96. H pylori status was assessed by presence of serum IgG antibodies. FINDINGS: 18 (8.0%) children at age 1-3 years had H pylori antibodies (13% black vs 4% white children, p=0.008). By age 18-23 years, the prevalence of the infection was 24.5% (43% black vs 8% white participants, p< 0.0001). Of the 206 children not infected at baseline, 40 (19%) became infected by age 21-23. The crude incidence rate per year was 1.4% for the whole cohort, ranging from 2.1% at 4-5 years and 1.5% at age 7-9 years to 0.3% at 21-23 years. The seroconversion rate was higher among black than among white children (relative risk 3.3, 95% CI 1.8-6.2, p=0.001). The median age for seroconversion was 7.5 years for both races. Nine of the 58 seropositive children cleared the infection during follow-up. The rate of seroreversion per year was 1.1%; it was highest among children at age 4-5 years (2.2% vs 0.2% at ages 18-19). INTERPRETATION: Most newly acquired H pylori infections happened before age 10 years. Treatment and preventive strategies should be aimed at children in this age-group. PMID- 11918914 TI - Crash diet potomania. PMID- 11918913 TI - Molecular remodelling of dystrophin in patients with end-stage cardiomyopathies and reversal in patients on assistance-device therapy. AB - BACKGROUND: Mutations that lead to disruption of cytoskeletal proteins have been recorded in patients with familial dilated cardiomyopathy. We postulated that changes in cytoskeletal and sarcolemmal proteins provide a final common pathway for dilation and contractile dysfunction in dilated cardiomyopathy. In this study, we investigated the integrity of dystrophin in the myocardium of patients with end-stage heart failure due to ischaemic or dilated cardiomyopathy, and the response to treatment with left-ventricular assistance devices (LVAD). METHODS: We assessed the expression and integrity of dystrophin in myocardial biopsy samples by immunohistochemistry and western-blot analysis using antibodies against the amino-terminal, carboxyl-terminal, and midrod domains. We took samples from the myocardia of ten controls, ten patients with dilated cardiomyopathy, ten with ischaemic heart disease, and six with dilated cardiomyopathy who underwent placement of a left-ventricular assistance device for progressive refractory heart failure. FINDINGS: Immunohistochemical staining identified a disruption to the amino-terminus of dystrophin in 18 of 20 patients with end-stage cardiomyopathy (dilated or ischaemic), whereas staining with antibodies against other domains of dystrophin was normal. Western-blot analysis confirmed these observations, suggesting that remodelling of dystrophin is a common pathway for dysfunction of failing cardiomyocytes. Furthermore, this disruption was reversible in four patients after LVAD support. INTERPRETATION: Dystrophin remodelling is a useful indicator of left-ventricular function in patients with dilated and ischaemic cardiomyopathy. Our results lend support to the hypothesis that changes in cytoskeletal proteins and, in particular, dystrophin might provide a final common pathway for contractile dysfunction in heart failure and these changes might be reversible by reduction of mechanical stress. PMID- 11918915 TI - Co-expression of survivin and TERT and risk of tumour-related death in patients with soft-tissue sarcoma. AB - Increased expression of survivin has been shown to be a negative predictor of survival in patients with soft-tissue sarcoma. We investigated 89 adults with soft-tissue sarcomas to ascertain the relation between co-expression of survivin and human telomerase reverse transcriptase (TERT) transcripts and prognosis. We quantified mRNA expression of survivin and TERT transcripts. Cox's proportional hazards regression model showed co-expression of both genes to be a significant negative prognostic factor for patients with stage I to stage IV tumours (p=0 small middle dot0004; relative risk 20 small middle dot1, 95% CI 3 small middle dot8-106 small middle dot4) and for those at stage II and III (p=0 small middle dot0002; 42 small middle dot1, 6 small middle dot0-294 small middle dot9) compared with low expression of both genes. Co-expression of survivin and TERT transcripts identifies patients at high risk of tumour-related death. PMID- 11918916 TI - One-step immunochromatographic assay for screening of coeliac disease. AB - Tissue transglutaminase is the autoantigen that elicits endomysial antibodies, which are the serological hallmarks of coeliac disease. We describe a simple, rapid immunochromatographic assay for IgA and IgG antibodies to transglutaminase, which is highly accurate for diagnosis of this disease. Results were positive for all samples from 50 untreated coeliac patients, and negative for 40 non-coeliac patients with gastrointestinal disorders. The assay seems to be a useful alternative to biopsy for mass screening for coeliac disease. PMID- 11918917 TI - Autologous thrombin for treatment of pseudoaneurysms. AB - Femoral pseudoaneurysms arise in up to 2% of patients after femoral cannulation for cardiac catheterisation. We used autologous thrombin for percutaneous obliteration of pseudoaneurysms occurring after catheterisation. We prepared autologous thrombin isolates from blood of ten patients with femoral pseudoaneurysms, and injected this solution into the pseudoaneurysms with duplex imaging guidance. We then assayed thrombin activity. All pseudoaneurysms were successfully thrombosed without substantial complications, although three patients needed a repeat procedure within 24 h. We have shown that autologous thrombin-induced thrombosis of pseudoaneurysms is reliable, simple, safe, and cheaper than commercial bovine or human thrombin, and avoids risks of anaphylaxis and contamination with prions. PMID- 11918918 TI - Effect of airborne allergens on emergency visits by children for conjunctivitis and rhinitis. AB - The effect of fungal spores and pollen grains on morbidity from childhood conjunctivitis and rhinitis is mostly unknown. We therefore studied the association between daily concentrations of these airborne allergens and daily emergency visits to a children hospital between 1993 and 1997. An increase of 551 basidiomycetes spores per m(3), or of 72 ragweed grains per m(3), was associated with an increase of about 10% in visits for conjunctivitis and rhinitis (p<0.01). Our results suggest that conjunctivitis and rhinitis could be caused by fungal spores and pollens in the air. PMID- 11918920 TI - Is gene-based therapy the way forward for spinal cord injury? PMID- 11918919 TI - Controversy surrounds proposed xenotransplant trial. PMID- 11918921 TI - Molecular shield that protects mucosal surfaces. PMID- 11918922 TI - Researchers question ability of stem cells to generate multiple cell types. PMID- 11918923 TI - Anthrax vaccine is safe and effective-but needs improvement, says IOM. PMID- 11918924 TI - Drosophila still flying high in cancer research. PMID- 11918925 TI - Mental health, truth, and justice in Guatemala. PMID- 11918926 TI - Croatia opens a national centre for the prevention of smoking. PMID- 11918927 TI - South African judge reaffirms judgement to expand access to AIDS drug. PMID- 11918931 TI - Africans discuss ethics of biomedical research. PMID- 11918929 TI - Bush to choose Johns Hopkins researcher for NIH top spot. PMID- 11918932 TI - Immunisations in solid-organ transplant recipients. AB - Solid-organ transplant recipients are at increased risk of various infectious diseases, some of which are vaccine preventable mmunisations are among the most efficient interventions available. Solid-organ tranplant recipients would greatly benefit from effective immunisations, provided the recommendations are based on a careful risk-benefit analysis in which the effectiveness of the vaccine is weighed against possible adverse reactions, including graft rejection. In this review, we summarise the data from studies on relevant immunisations in solid organ transplant recipients. The major issues are the immunogenicity and safety of immunisations, the factors associated with poor immune response, and recommendations for immunisation schemes. PMID- 11918933 TI - Unequal group sizes in randomised trials: guarding against guessing. AB - We cringe at the pervasive notion that a randomised trial needs to yield equal sample sizes in the comparison groups. Unfortunately, that conceptual misunderstanding can lead to bias by investigators who force equality, especially if by non-scientific means. In simple, unrestricted, randomised trials (analogous to repeated coin-tossing), the sizes of groups should indicate random variation. In other words, some discrepancy between the numbers in the comparison groups would be expected. The appeal of equal group sizes in a simple randomised controlled trial is cosmetic, not scientific. Moreover, other randomisation schemes, termed restricted randomisation, force equality by departing from simple randomisation. Forcing equal group sizes, however, potentially harms the unpredictability of treatment assignments, especially when using permuted-block randomisation in non-double-blinded trials. Diminished unpredictability can allow bias to creep into a trial. Overall, investigators underuse simple randomisation and overuse fixed-block randomisation. For non-double-blinded trials larger than 200 participants, investigators should use simple randomisation more often and accept moderate disparities in group sizes. Such unpredictability reflects the essence of randomness. We endorse the generation of mildly unequal group sizes and encourage an appreciation of such inequalities. For non-double-blinded randomised controlled trials with a sample size of less than 200 overall or within any principal stratum or subgroup, urn randomisation enhances unpredictability compared with blocking. A simpler alternative, our mixed randomisation approach, attains unpredictability within the context of the currently understood simple randomisation and permuted-block methods. Simple randomisation contributes the unpredictability whereas permuted-block randomisation contributes the balance, but avoids the perfect balance that can result in selection bias. PMID- 11918934 TI - The right to sell or buy a kidney: are we failing our patients? PMID- 11918935 TI - Need for caesarean sections in west Africa. PMID- 11918936 TI - Need for caesarean sections in west Africa. PMID- 11918937 TI - Need for caesarean sections in west Africa. PMID- 11918940 TI - Telomere shortening and atherosclerosis. PMID- 11918942 TI - Repair of coarctation: a higher goal? PMID- 11918943 TI - Fatal hepatic abscess caused by a fish bone. PMID- 11918944 TI - Interferon alfa-2a for melanoma metastases. PMID- 11918945 TI - Low socioeconomic status and coronary artery disease. PMID- 11918948 TI - Low socioeconomic status and coronary artery disease. PMID- 11918946 TI - Is radiation nephropathy caused by yttrium-90? PMID- 11918949 TI - False-negative food challenges. PMID- 11918950 TI - Medicine and self-image in literature. PMID- 11918951 TI - Medicine and self-image in literature. PMID- 11918952 TI - Medicine and self-image in literature. PMID- 11918953 TI - The deterrence issue. PMID- 11918954 TI - The deterrence issue. PMID- 11918964 TI - The chief's broken leg. PMID- 11918966 TI - Energy metabolism, stress hormones and neural recovery from cerebral ischemia/hypoxia. AB - All the advancements in the understanding of the molecular and cellular processes leading to the great investments in developing neuroprotection against cerebral ischemic/hypoxic damage cannot obscure the simple fact that exhaustion of energy supplies is still at the basis of this disorder. Much has been investigated and postulated over the years about the quick collapse of energy metabolism that follows oxygen and glucose deprivation in the brain. Anaerobic glycolysis, recognized as a pathway of paramount importance in keeping energy supplies, although, at bare minimum, has also presented a dilemma-a significant increase in lactate production during ischemia/hypoxia (IH). The dogma of lactate as a useless end product of anaerobic glycolysis and its postulated role as a detrimental player in the demise of the ischemic cell has persisted for the past quarter of a century. This persistence is due to, at least in part, the well documented phenomenon termed "the glucose paradox of cerebral ischemia," the unexplained aggravation of postischemic neuronal damage by preischemic hyperglycemia. Recent studies have questioned the deleterious effect of lactic acid, while others even have offered the possibility that this monocarboxylate serves as an aerobic energy substrate during recovery from IH. Reviewed here are studies published over the past few years along with some key older papers on the topic of energy metabolism and recovery of neural tissue from IH. New insights gained from both in vitro and in vivo studies on energy metabolism of the ischemic/hypoxic brain should improve our understanding of this key metabolic process and the chances of protecting this organ from the consequences of energy deprivation. PMID- 11918967 TI - Role of tyrosine, DOPA and decarboxylase enzymes in the synthesis of monoamines in the brain of the locust. AB - The metabolic transformation of tyrosine (TYR) by the decarboxylase and hydroxylase enzymes was investigated in the central nervous system of the locust, Locusta migratoria. It has been demonstrated that the key amino acids, 3,4 dihydroxyphenylalanine (DOPA), 5-hydroxytryptophan (5HTP) and tyrosine are decarboxylated in all part of central nervous system. DOPA and 5HTP decarboxylase activities show parallel changes in the different ganglia, but the rank order of the activity of TYR decarboxylase is different. Enzyme purification has revealed that the molecular weights of TYR decarboxylase and DOPA/5HTP decarboxylase are 370,000 and 112,000, respectively. The decarboxylation of DOPA by DOPA/5HTP decarboxylase is stimulated, whereas the decarboxylation of DOPA by TYR decarboxylase is inhibited in the presence of the cofactor pyridoxal-5' phosphate. TYR hydroxylase could not be detected and 3H-TYR is found to be metabolised to tyramine (TA), but not to DOPA. The haemolymph contains a significant concentration of DOPA (120 pmol/100 microl haemolymph), and the ganglia incorporates DOPA from the haemolymph by a high affinity uptake process (K(M)=12 microM and V(max)=24 pmol per ganglion/10 min). Our results suggest that no tyrosine hydroxylase is present in the locust CNS and the DOPA uptake into the ganglia by a high affinity uptake process as well as the DOPA decarboxylase enzyme may be responsible for the regulation of the ganglionic dopamine (DA) level. Two types of decarboxylases exist, one of them decarboxylating DOPA and 5HTP (DOPA/5HTP decarboxylase), other decarboxylating TYR (TYR decarboxylase). The DOPA/5HTP decarboxylase enzyme present in the insect brain may correspond to the 5HTP/DOPA decarboxylase in vertebrate brain, whereas TYR decarboxylase is characteristic only for the insect brain. PMID- 11918970 TI - TAT-mediated delivery of human glutamate dehydrogenase into PC12 cells. AB - Human glutamate dehydrogenase (GDH) gene was fused with a gene fragment encoding the nine amino acid (RKKRRQRRR) protein transduction domain of human immunodeficiency virus TAT protein in bacterial expression vector to produce genetic in-frame TAT-GDH fusion protein. The TAT-GDH protein can enter PC12 cells efficiently when added exogenously in culture media as determined by Western blot analysis and enzyme activities. Once inside the cells, the transduced denatured TAT-GDH protein showed a full activity of GDH indicating that the TAT-GDH fusion protein was correctly refolded after delivery into cells and the activities of GDH in the TAT-GDH fusion protein was not affected by the addition of the TAT sequence. TAT-GDH fusion protein and TAT itself showed no cytotoxicity in PC12 cells. Although the exact mechanism of transduction across a membrane remains unclear, the transduction activity of TAT-GDH into PC12 cells may suggest new possibilities for direct delivery of GDH into the patients with the GDH-deficient disorders. PMID- 11918969 TI - Effects of different oxidizing agents on neutral amino acid transport systems in isolated bovine brain microvessels. AB - Using isolated bovine brain microvessels as an in vitro model of the blood-brain barrier (BBB) we have evaluated the role of free radical generating solutions on some amino acid transport systems operating on the endothelial cell membrane. Fe(2+)/ascorbate, phenylhydrazine and CuSO(4) did not affect any of the transport system tested, while exposure of bovine brain microvessels to tert butylhydroperoxide (t-BHP) caused a reduced capacity to take up small neutral amino acids via the Na(+)-dependent A-system. The presence of glucose during t BHP treatment did not prevent this inhibition, which was partially counteracted when the isolated microvessels were incubated with 5mM inosine before the oxidative stress. Incubation of the isolated capillaries with 5mM dithiothreitol, after exposure to t-BHP, resulted in a 50% recovery of the alpha methylaminoisobutyrate (MeAIB) uptake by the A-system. Treatment with t-BHP, which had no effect on the L-system of neutral amino acid transport, caused a significant decrease of the intracellular levels of ATP, of glutathione (GSH), and of gamma-glutamyltranspeptidase (GGT) activity, while no significant modification of hexokinase (HK) or of alkaline phosphatase (ALKP) activities were observed. Oxidative damage of the BBB appears therefore to impair essentially the metabolic pathways which ensure the energy requirement for the endothelial cells, thus inhibiting the energy-dependent amino acid transport system "A". PMID- 11918968 TI - Synthesis and evaluation of (S)-[18F]-fluoroethylcarazolol for in vivo beta adrenoceptor imaging in the brain. AB - The beta-adrenergic receptor ligand (S)-4-(3-(2'-[18F]-fluoroethylamino)-2 hydroxypropoxy)-carbazol ((S)-[18F]-fluoroethylcarazolol) was prepared by reaction of [18F]-fluoroethylamine with the corresponding (S)-epoxide and was evaluated in rats by studying its pharmacokinetics and its binding profile both in vitro and in vivo. In vitro, (S)-fluoroethylcarazolol binds preferentially to beta-adrenoceptors (pK(i)=9.3 for beta(1) and 9.4 for beta(2)) and has less affinity to 5HT(1A) and 5HT(1D) receptors (pK(i)=6.7 and 5.2). In vivo, standard uptake values (SUVs) up to 0.63+/-0.07 in cortical regions were found after 60 min. Metabolites (90%) appeared within 10 min in plasma, whereas, in brain 70-75% parent compound was found after 60 min. Clearance from plasma occurred within 5 min. Cerebral uptake could be blocked by 'cold' fluoroethylcarazolol in every region, except medulla. Uptake was also blocked by propranolol and pindolol, but not by WAY 100635. ICI 89406 hardly lowered [18F] levels in brain. ICI 118551 reduced uptake of [18F] in cerebellum (mainly beta(2)) by 30%. Specific binding (tissue minus medulla values) in various brain regions corresponded with those observed for [18F]-fluorocarazolol (r(2)=0.95) and with in vitro beta adrenoceptor densities (r(2)=0.76). Autoradiography using phosphor images of (S) [18F]-fluoroethylcarazolol in rat brain showed the characteristic binding pattern of beta-antagonists, while propranolol treatment resulted in low and homogenous uptake. Regional tissue minus medulla values corresponded with in vitro beta adrenoceptor densities (r(2)=0.77). We conclude that (S)-[18F] fluoroethylcarazolol is a high affinity ligand that binds specifically to cerebral beta-adrenoceptors in vivo and may be of use for beta-adrenoceptor imaging in the brain with PET. PMID- 11918971 TI - Acetaminophen protects hippocampal neurons and PC12 cultures from amyloid beta peptides induced oxidative stress and reduces NF-kappaB activation. AB - The present findings show that an atypical non-steroidal anti-inflammatory drug, such as acetaminophen, retains the ability to recover amyloid beta-peptides driven neuronal apoptosis through the impairment of oxidative stress. Moreover, this compound reduces the increased NF-kappaB binding activity, which occurs in these degenerative conditions. Therapeutic interventions aimed at reducing the inflammatory response in Alzheimer's disease (AD) recently suggested the application of non-steroidal anti-inflammatory drugs. Although the anti inflammatory properties of acetaminophen are controversial, it emerged that in an amyloid-driven astrocytoma cell degeneration model acetaminophen proved to be effective. On these bases, we analyzed the role of acetaminophen against the toxicity exerted by different Abeta-peptides on rat primary hippocampal neurons and on a rat pheochromocytoma cell line. We found a consistent protection from amyloid beta-fragments 1-40 and 1-42-induced impairment of mitochondrial redox activity on both cell cultures, associated with a marked reduction of apoptotic nuclear fragmentation. An antioxidant component of the protective activity emerged from the analysis of the reduction of phospholipid peroxidation, and also from a significant reduction of cytoplasmic accumulation of peroxides in the pheochromocytoma cell line. Moreover, activation of NF-kappaB by amyloid-derived peptides was greatly impaired by acetaminophen pre-treatment in hippocampal cells. This evidence points out antioxidant and anti-transcriptional properties of acetaminophen besides the known capability to interfere with inflammation within the central nervous system, and suggests that it can be exploited as a possible therapeutic approach against AD. PMID- 11918972 TI - Expression in E. coli and purification of recombinant fragments of wild type and mutant human prion protein. AB - Prion diseases are fatal neurodegenerative disorders of the CNS of men and animals, characterized by spongiform degeneration of the CNS, astrogliosis and deposition of amyloid into the brain. The conversion of a cellular glycoprotein (the prion protein, PrP(C)) into an altered isoform (the prion scrapie, PrP(Sc)), which accumulates within the brain tissue by virtue of its resistance to the intracellular catabolism, is currently believed to represent the etiologic agent responsible for these diseases. Synthetic or recombinant polypeptides are commonly used to elucidate the mechanism of proteins involved in neurodegenerative diseases. Here we describe a procedure, which allows the synthesis and purification in its native folding, of the human prion protein fragment 90-231, corresponding to the protease resistant core of PrP(Sc). We synthesized the polypeptides 90-231 of both the wild type and the E200K mutant isoforms of PrP. Using a gluthatione S-transferase (GST) fusion protein approach, milligram amounts of polypeptides were obtained after expression in E. coli. The recovery of the purified fusion protein was monitored following the evaluation of the GST activity. The PrP fragment was released from the fusion protein immobilized on a glutathione-coupled agarose resin by direct cleavage with thrombin. The recombinant protein was identified by comassie stained acrylamide gel and by immunoblotting employing a monoclonal anti-PrP antibody. The peptide purified by gel filtration chromatography showed mainly an alpha-helix structure, as analysed by circular dichroism (CD) and an intact disulfide bridge. The same procedure was also successfully employed to synthesize and purify the E200K mutant PrP fragment. PMID- 11918973 TI - Distribution of neuronal and non-neuronal spliced variants of type 1 IP(3) receptor in rat hypothalamus and brain stem. AB - In the nervous system, inositol 1,4,5-trisphosphate (IP(3)) is one of the second messengers produced by PI hydrolysis and triggers IP(3)-receptor (IP(3)R) mediated calcium release from intracellular pools. Throughout the brain, the type 1 IP(3)R is predominantly expressed and its mRNA is widely distributed. Alternative splicing of IP(3)R1 (SI and SII) occurs in two distinct regions. SI splicing in the middle of the ligand binding domain may alter the IP(3) binding activity, while SII splicing probably affects the protein kinase A phosphorylation sites and kinetics. Selective use of IP(3)-receptor subtypes may permit a tissue specific and developmentally specific expression of functionally distinct channels. The present work was focused on detection of the alternatively spliced mRNA of type 1 IP(3)-receptor in individual brain structures and nuclei. Using RT-PCR we detected neuronal (535bp) and non-neuronal (410bp) forms. We identified both spliced variants in the majority of brain structures, except in the cerebellum and medulla. In the cerebellum, the neuronal form of type 1 IP(3)R was found exclusively, while in the medulla, the non-neuronal form was much more abundant. Nevertheless, Western blot analysis and hybridization with specific antibody against IP(3)R revealed no qualitative, but only quantitative differences. Similarly, IP(3) dependent calcium release did not show any differences between the cerebellum and pons. These results demonstrate the distribution of alternatively spliced S2 variants of type 1 IP(3)R in selected brain structures and nuclei. The physiological relevance of these two forms remains to be elucidated by further studies. PMID- 11918974 TI - Reduced pteridine derivatives induce apoptosis in PC12 cells. AB - In cerebrospinal fluid of patients with cerebral infections, elevated concentrations of the pteridine compounds neopterin and 7,8-dihydroneopterin were detected. Here, the potential of pteridines to induce apoptosis of the rat pheochromocytoma cells (PC12) was investigated. In contrast to aromatic pteridines like neopterin, the reduced forms 7,8-dihydroneopterin, 5,6,7,8 tetrahydrobiopterin and 7,8-dihydrobiopterin led to a significant increase of apoptotic cells. After terminal differentiation, cells were less sensitive to incubation with pteridines. A noticeable augmentation of apoptosis was observed upon incubation with 7,8-dihydroneopterin and 7,8-dihydrofolic acid. Antioxidants partly protected PC12 cells from pteridine-induced apoptosis, suggesting the involvement of reactive oxygen intermediates. Exposure of cells to 7,8 dihydroneopterin led to activation of the mitogen-activated protein (MAP) kinase and to a lesser degree also of JUN/SAP kinase. Results implicate that high concentrations of reduced pteridines, might contribute to the pathogenesis involved in neurodegeneration. PMID- 11918975 TI - An assisted automated procedure for vessel geometry reconstruction and hemodynamic simulations from clinical imaging. AB - In this paper a method is described to obtain realistic 3-D geometric models of vascular districts from clinical tomographic 3-D images. The aim is the simulation of individual local hemodynamics by means of computational fluid dynamics (CFD). As a test case, the method is applied to the carotid bifurcation. Attention is focused on the minimisation of the time demanding costs. The proposed procedure has been automated whenever possible and takes about 2h from the acquisition of the images to the attainment of the simulation results, a time lapse compatible with diagnostic exigency. PMID- 11918976 TI - Comparison of flat-panel detector and image-intensifier detector for cone-beam CT. AB - We evaluated a flat-panel detector (FPD) (scintillator screen and a-Si photo sensor array) for use in a cone-beam computed tomography (CT) detector and compared it with an image-intensifier detector (IID). The FPD cone-beam CT system has a higher spatial resolution than the IID system. At equal pixel sizes, the standard deviation of noise intensity of the FPD system is equal to that of the IID system. However, the circuit noise of the FPD must be reduced, especially at low doses. Our evaluations show that the FPD system has a strong potential for use as a cone-beam CT detector because of high-spatial resolution. PMID- 11918977 TI - Fat-suppressed 3D-T1-weighted-echo planar imaging: comparison with fat-suppressed 3D-T1-weighted-gradient echo in imaging the cartilage of the knee. AB - This study was conducted to compare a three-dimensional (3D) multi-shot echo planar imaging (EPI) sequence with fat-suppression (FS) with the 3D-fat suppressed gradient echo (GRE-FS) sequence in imaging the cartilage of the knee. One hundred sixty-nine patients were studied prospectively. The cartilage was imaged in the sagittal plane with: (a) 3D-T1-EPI-FS and (b) 3D-T1-GRE-FS sequences using a 1T MR scanner. The signal-to-noise ratio (SNR) of bone (b) and cartilage (c), and relative contrast (ReCon) between bone and cartilage and meniscus and cartilage were measured in 60 patients with arthroscopically normal cartilage. The imaging accuracy was assessed by comparing with linear regression analysis (length and depth) 32 defects in the cartilage of cadaveric (human and bovine) knees. The 3D-T1-EPI-FS provided better bone marrow signal suppression, better SNRc and better ReCon(bc) and ReCon(cm) (p<0.01). The 3D-T1-EPI-FS showed better accuracy concerning the depth of the defects and the 3D-T1-GRE-FS better accuracy concerning the length of the defects. In conclusion, the 3D-T1-EPI-FS pulse sequence could be included in the routine protocol in imaging the cartilage of the knee because it achieves high SNR of the cartilage and high ReCon compared to the surrounding structures, at a reduced scan time. PMID- 11918978 TI - Stromal predominant type mesenchymal hamartoma of liver: CT and MR features. AB - Mesenchymal hamartoma is a rare pediatric liver tumor. The tumor is mainly cystic but occasionally a solid component is seen. Therefore the tumor can be divided into two forms: (a) cystic predominant, and (b) stromal predominant. In this report, CT and MR features of stromal predominant type mesenchymal hamartoma are presented. PMID- 11918979 TI - The accuracy of three-dimensional reconstructions of the ovine knee: dissectional validation. AB - A single, complete, ovine knee was imaged using conventional MRI then dissected allowing comprehensive dimensional measurement of the intra-articular structures. A three-dimensional (3-D) computer model of the knee was generated from the MR sections. Thirty-two individual structural measures were recorded from the image output for which there was an available surgical measure for direct comparison. The results of Pearson's correlation testing show a rounded score of 1.00, suggesting an exceptional linear correlation between direct anatomical measurement and the 3-D image output. Further analysis of the data revealed an average error of measurement of 0.2mm across the 32 measures. The findings of this preliminary study suggest that 3-D reconstruction from MR data may be an appropriate, and accurate, means for making dimensional measurements of the bony and soft tissue structures of the ovine knee. It is unlikely that the measurement error would be of any great clinical significance. There is evidence in the literature to suggest that an ovine knee may be considered an acceptable model for substitution for the human knee in diagnostic assessment studies. Therefore, such findings may be considered clinically relevant in the field of human knee assessment. PMID- 11918980 TI - Ectopic spleen and left-sided vena cava in Beckwith-Wiedemann syndrome. AB - Beckwith-Wiedemann syndrome (BWS) is a congenital overgrowth syndrome characterized by anterior abdominal wall defects, macroglossia, and gigantism. A variety of other abnormalities have been described, however association with ectopic spleen and left-sided vena cava has not been reported previously. We report ectopic spleen, left-sided vena cava and the other abdominal imaging findings of an adult BWS case who came up to date without any follow-up from the early childhood. PMID- 11918981 TI - CRT diagnosis of pulmonary disease: influence of monitor brightness and room illuminance on observer performance. AB - Using a 21-in. cathode ray tube (CRT) monitor (2048 x 2560 x 8bits), six radiologists interpreted 12 images with interstitial lung disease under six conditions of CRT luminance (50 and 400 cd/m(2)) and room illuminance (20, 120 and 480lx), and 10 radiologists interpreted 25 images with pulmonary nodules under nine conditions of CRT luminance (50, 200 and 500 cd/m(2)) and room illuminance (20, 120 and 480lx). Observer's performance for interstitial disease was relatively better at 120lx. Four hundred and eighty lux illuminance with 50 cd/m(2) CRT luminance, which degraded the detectability of pulmonary nodule significantly (p<0.05), should be avoided for clinical use. PMID- 11918982 TI - Astroblastoma: diffusion MRI, and proton MR spectroscopy. AB - A patient with grade II astroblastoma is reported. The tumor revealed an inhomogenous enhancement pattern on T1-weighted images after contrast medium. It contained cyst-like structures with high signal in FLAIR images, consistent with presence of viscous material. In spectroscopy, NAA was decreased, and Cho was increased. Peaks belonging to lipids and a combined broad peak belonging to myoinositol and glycine were noticed. In diffusion MRI, the mean apparent diffusion coefficient (ADC) value of intratumoral cyst-like structures was 2.12 x 10(-3)mm(2)/s less than that of CSF. The matrix of the tumor had lower ADC values ranging from 1.19 to 1.25 x 10(-3)mm(2)/s, higher than that of normal cerebral parenchyma. PMID- 11918983 TI - Ovarian tumor with functioning stroma. AB - The authors reported two cases with ovarian tumors with functioning stroma. One was an ovarian carcinoid with testosterone-producing stroma and another was an ovarian mucinous cystadenoma with elevated serum level of estrogen. They appeared as multilocular cystic masses with varying signal on MR. This imaging feature was mimicking both those of Sertoli-Leydig tumors with heterologous elements and granulosa cell tumors, which are well known as functioning tumors. It had been reported in pathological literatures that any type of ovarian tumors can show hormonal activity due to functional stroma, although, it is the first report in the radiological literatures. PMID- 11918984 TI - Huge craniopharyngioma: diffusion MRI and contrast-enhanced FLAIR imaging. AB - Craniopharyngiomas are most commonly located extraaxially in the suprasellar area. They are benign but aggressive neoplasms. An adult patient is reported to have a huge craniopharyngioma with gross extensions to the surroundings. In diffusion MRI, it had high signal for b=1000 mm(2)/s (true diffusion) images, and at the same time, high ADC values (=2.12 and 2.27 x 10(-3)mm(2)/s, compared to that of normal cerebellar parenchyma 0.85 x 10(-3)mm(2)/s). In FLAIR images, obtained after administration of intravenous contrast medium, an intense, diffuse enhancement pattern was seen involving the viable tumor portions as well as the intratumoral fluid. PMID- 11918985 TI - Dissection of bilateral internal carotid arteries and occlusion of both vertebral arteries in a child patient. PMID- 11918986 TI - Alcohol dependence and gender-role orientation. AB - OBJECTIVE: The overall increase of female alcoholism is supposed to be associated with the change of the traditional female role, and it is especially seen as a consequence of role convergence or gender-role conflicts. The aim of the present pilot study is to explore whether the approach of gender-role orientation would be empirically useful in contributing to these hypotheses. METHOD: One hundred twelve patients with alcohol dependence meeting DSM-III-R criteria were explored after detoxification; gender-role orientation was measured by a German version of the "Extended Personal Attributes Questionnaire", categorising gender-role orientation into four subgroups: masculine, feminine, androgynous, and undifferentiated. RESULTS: In comparison with a population-based sample, there are significant differences in the distribution of the four subgroups of gender role orientation, showing a predominance of the undifferentiated self-concept in the alcoholic sample (49%). Alcoholic females describe themselves as rather undifferentiated, and rather feminine than masculine. Low masculinity and low femininity, as well as high femininity, correlate positively with distress, depressiveness, social anxiety, insecurity and concomitant personality disorders. CONCLUSION: Our data do not support the convergence hypothesis related to gender role orientation, but support the traditional feminine self-concept as an unspecific risk factor for vulnerability. The question whether an undifferentiated self-concept could be a specific risk factor for alcoholism is discussed. PMID- 11918987 TI - Dopaminergic dysfunction in alcoholism and schizophrenia--psychopathological and behavioral correlates. AB - Dysfunction of central dopaminergic neurotransmission has been implicated in the pathogenesis of schizophrenia as well as drug and alcohol dependence. Different drugs of abuse stimulate dopamine release in the ventral striatum and thus reinforce drug consumption. Increased subcortical dopamine release has also been associated with the pathogenesis of positive symptoms in schizophrenia and may be driven by a prefrontal dopaminergic dysfunction. These seemingly heterogeneous findings may be explained by recent research in non-human primates. According to these studies, reward anticipation but not anticipated reward consumption is accompanied by a phasic dopamine release in the striatum and prefrontal cortex. In the striatum, phasic dopamine release primarily affects motivation, psychomotor activation and reward craving, while in the prefrontal cortex, dopaminergic stimulation is involved in the activation of working memory and reward anticipation. In alcoholism, previously neutral stimuli that have been associated with alcohol intake can become conditioned cues which activate phasic dopamine release and reward craving. In schizophrenia, stress-induced or chaotic activation of dopamine release may attribute incentive salience to otherwise irrelevant stimuli and thus be involved in the pathogenesis of delusional mood and other positive symptoms. Studies in humans and non-human primates emphasize the role of dopaminergic neurotransmission in reward anticipation and its dysfunction in different neuropsychiatric diseases. PMID- 11918988 TI - D(2) dopamine receptor (DRD2) polymorphism is associated with severity of alcohol dependence. AB - The A(1) allele of the D(2) dopamine receptor (DRD2) gene has been associated with alcohol dependence. However, the expression of this allele risk on the severity of drinking behavior in patients with alcohol dependence has not been systematically explored. The present study examines the association between DRD2 A(1)(+) (A(1)/A(1) and A(1)/A(2) genotypes) and A(1)- (A(2)/A(2) genotype) allele status and key drinking parameters in alcohol-dependent patients. A sample of Caucasian adults was recruited from an alcohol detoxification unit. A clinical interview and the Alcohol Dependence Scale (ADS) questionnaire provided data on consumption, dependence, chronology of drinking and prior detoxification. A(1)(+) allele compared to A(1)- allele patients consumed higher quantities of alcohol, commenced problem drinking at an earlier age, experienced a shorter latency between first introduction to alcohol to the onset of problem drinking and had higher ADS scores. Moreover, A(1)(+) allele patients had more detoxification attempts than their A(1)- allele counterparts. In sum, alcohol-dependent patients with the DRD2 A(1) allele compared to patients without this allele are characterized by greater severity of their disorder across a range of problem drinking indices. The implications of these findings are discussed. PMID- 11918989 TI - 5HTR2A gene polymorphism and personality traits in patients with major psychoses. AB - Serotonin receptor (5HTR2A) gene polymorphism has been reported to be associated with clinical phenotypes in schizophrenia. The current study attempted to investigate a relationship between 5HTR2A 102T/C polymorphism and personality traits as well as clinical symptoms in patients with ICD-10 diagnoses of schizophrenia and affective disorders. 5HTR2A genotyping, clinical and psychological assessment were administered to 375 patients, 104 first-degree healthy relatives of the patients and 157 controls. In the patients an association was observed between the 2/2 5HTR2A genotype and scores on the Hypochondriasis scale (MMPI) (ANOVA, F = 4.56; P = 0.011) and trait anxiety (F = 4.21; P = 0.002). A significant difference between 1/1 and 2/2 genotypes has been also found for Neuroticism scores (EPI) (t = 2.18; P = 0.0031). No significant differences by 5HTR2A genotype were observed in either the control or first degree relatives group for all scales studied. Positive, negative and psychopathological symptoms emerged higher in the 2/2 genotype patients compared to other genotype carriers. Therefore, the 2/2 genotype may contribute to produce the phenotype, with specific clinical and pathological features in common, regardless of nosologic heterogeneity of psychoses. PMID- 11918990 TI - Gender differences in panic disorder symptoms and illicit drug use among young people in Hungary. AB - The authors have investigated the frequency of illicit drug use, and spontaneous and drug-provoked panic disorder symptoms in Hungarian youths. A semi-structured self-reporting questionnaire (with questions about drug usage and the persistence of some DSM-IV panic disorder symptoms) was filled out in discos/nightclubs, secondary schools and universities. Almost 17% of the total sample (n = 1298) reported on illicit drug-use at least once in their life, and this rate was significantly higher among males. Regardless of the illegal drug use 14.6% of the total sample reported on four or more DSM-IV symptoms of panic disorder, and this rate was significantly higher among females. Analyzing the panic disorder symptoms only among drug-users (n = 219), the frequency of persons with four or more anxiety symptoms was 14.1% before drug use, and it increased to 30.6% during the period of drug use (P < 0.001). The findings support previous results showing (i) higher prevalence of anxiety symptoms among females; (ii) higher rate of illicit drug use among males; and (iii) a possible anxiety-provoking effect of illicit drugs. PMID- 11918991 TI - Predictors of improvement in quality of life of long-term mentally ill individuals receiving case management. AB - One hundred and thirteen long-term mentally ill clients receiving case management were investigated with regard to psychosocial and clinical predictors of changes in subjective quality of life during an 18-month follow-up. Better psychosocial functioning and fewer psychiatric symptoms at baseline predicted a greater improvement in quality of life. A larger decrease in symptom severity and a greater improvement in the social network during the follow-up were identified as the most important predictors of a greater improvement in subjective quality of life. The results of the study suggest that an emphasis should be put on effective symptom management, a reduction of needs for care and social support in order to fulfill the aims of improving subjective quality of life in patients receiving case management. PMID- 11918992 TI - Barriers to the use of electroconvulsive therapy in the elderly: a European survey. AB - A postal survey was carried out to determine the clinical and legal guidelines governing the use of electroconvulsive therapy (ECT) in the countries of the wider Europe. Respondents from 23 of the 33 countries returned completed questionnaires. Considerable variation was found in the availability of ECT, the frequency of its use and associated legal procedures. However, there was a broad consensus with regard to the clinical indications. Access to the treatment was most frequently limited by financial or other resource constraints, political or legal restrictions. PMID- 11918993 TI - A blind spot for secondary gain affecting therapy outcomes. AB - BACKGROUND: A study to determine whether the expectation for secondary gain held by psychiatric outpatients was associated with therapy outcomes. METHODS: The study was conducted at a Dutch psychiatric outpatient department. Our investigation set out to explore the expectation of obtaining secondary gain by directly asking the patient and clinician via questionnaires whether the patient anticipated to get specific "benefits" from being in therapy. RESULTS: From 166 patients, 70 (42.2%) reported to expect secondary gain while in therapy. We found a significant relation between expectation for secondary gain and treatment outcomes. Patients with expectations for secondary gain were significantly more prone to poor therapy outcome. Only in nine of 147 cases (6%) did patients explicitly express their expectations for secondary gain towards a psychiatrist. Moreover, expectations for secondary gain did not appear to be related to Axis I and Axis II diagnoses. DISCUSSION: Secondary gain appeared to be a "veiled motive" for getting therapy: patients did not express their expectations for secondary gain explicitly towards a psychiatrist. This aspect of veiled motives is of particular relevance in regard to the fact that expectations for secondary gain appeared to affect therapy results. PMID- 11918994 TI - Myoclonic and generalized tonic clonic seizures during combined treatment with low doses of clozapine and haloperidol. PMID- 11918995 TI - Collective bursting in layer IV. Synchronization by small thalamic inputs and recurrent connections. AB - Layer IV is believed to be the cortical signal amplifier, for example, of thalamic signals. A previous spiny stellate recurrent network model of this layer is made more realistic by the addition of inhibitory basket neurons. We study the persistence and characteristics of previously observed collective firing behavior, and investigate what additional features would need to be implemented to generate in vivo type neuronal firing. It is shown that neuronal activity is only coarsely synchronized within the network. By applying methods of noise cleaning, it emerges that the firing of individual neurons is of low-dimensional hyperchaotic nature, as found in the analysis of measured cat in vivo spike trains. In order to reproduce in vivo firing patterns, it is sufficient to have time-varying thalamic input. Conclusions from low-dimensional hyperchaotic behavior of network-embedded neurons are drawn. We interpret observed in vivo pattern-sharpening features of stimuli and outline possible connections to epilepsy. From our results, it follows that emergent global behavior is likely to be the result of the interaction between comparably simple neuronal components, driven by input specificity. PMID- 11918996 TI - No effect of gender on tonal and phonetic mismatch negativity in normal adults assessed by a high-resolution EEG recording. AB - The auditory mismatch negativity (MMN) of event-related potential components has been widely used to assess the ability of auditory automatic change discrimination of verbal and nonverbal stimuli in healthy individuals and patients with various illnesses. To clarify the role of gender differences in the MMN, we compared the amplitude, latency, and topography of tonal and phonetic MMN between healthy males and females, using a high-density (128 channel) electroencephalography montage. The MMN was evaluated in 18 right-handed male and ten age-matched female adults. The MMN in response to a duration change of pure tone and that in response to a phonetic change (Japanese vowel /a/ versus /o/ with 150-ms duration) were recorded. There were no significant differences in amplitude, latency, or laterality for either tonal or phonetic MMN between male and female subjects. This lack of evidence for effects of gender on MMN in response to duration change of tones or that in response to changes of phonemes with a short duration in normal adults may be of relevance to a growing number of researchers who are studying the MMN in healthy individuals and various clinical groups. PMID- 11918997 TI - From primed to learn: the saturation of repetition priming and the induction of long-term memory. AB - Although practice can make perfect, it is not clear how much practice is needed to trigger long-lasting performance gains on a given task. Here, using a letter enumeration task, we show that the transition of experience dependent performance gains to a relatively stable form, as well as the triggering of delayed, long lasting, between session gains (both effects are considered manifestations of consolidation processes) is amount-of-practice dependent. We then show (a) that consolidation processes, once triggered, can proceed without further practice as a function of time and (b) that the triggering of consolidation processes is related to repetition priming effects--performance gains in processing a previously experienced item. However, we show that repetition priming effects saturate after a limited number of consecutive repetitions and reflect an initial, but potentially reversible, response to the repeated experience. Moreover, we show that one critical parameter determining the occurrence of repetition priming (but not skill learning) is the presence of interference (by a somewhat different set of items) prior to the primer presentation. Thus, our results suggest that the saturation of repetition priming effects, rather than priming per se, may be critical to the induction of slow learning processes and consolidation. PMID- 11918999 TI - Differential task effects on semantic and syntactic processes as revealed by ERPs. AB - Two experiments investigated the time-course of semantic and syntactic processes in auditory language comprehension as well as their possible functional dependencies, using event-related brain potentials (ERPs). Participants listened to sentences which were either correct, semantically incorrect, syntactically incorrect, or both semantically and syntactically incorrect. In experiment 1, participants judged the overall correctness of these sentences. The semantic violation elicited an N400 whereas the syntactic phrase structure violation elicited an early anterior negativity followed by a P600. Sentences in which the critical element violated both semantic and syntactic constraints elicited the same pattern of ERPs as the syntactic violation alone, not evoking an N400. In experiment 2, participants judged the same sentences for semantic coherence, required to ignore syntactic violations. Again, an early anterior negativity was elicited for those sentences containing phrase-structure errors. In contrast to experiment 1, however, combined violations elicited both an early negativity and an N400. Together, the results suggest that the N400 associated with semantic aspects of sentence comprehension reflects controlled processes whereas initial parsing processes associated with the early anterior negativity are independent of semantic constraints and task requirements. PMID- 11918998 TI - Age-related changes in source memory retrieval: an ERP replication and extension. AB - Event-related potentials (ERPs) were recorded (62 scalp sites) from young (M=22) and older (M=66) adults during tests of item recognition and source memory, in a replication and extension of a previous study [Psychol. Aging 14 (1999) 390-413]. Participants studied two temporally distinct lists of sentences (each with two unassociated nouns). At test, in response to studied and unstudied nouns, participants made old/new, followed by source (i.e., list) judgments. Several measures were employed to enhance the source memory performance of the older adults. These were successful, as the old adults showed source memory performance comparable to that of the young subjects from the previous study (67%). Nonetheless, the younger adults significantly outperformed the older adults on measures of item and source memory performance. The ERPs revealed that both age groups showed a robust early, posterior-maximal episodic memory (EM) effect. However, despite their enhanced source memory performance, the ERPs of the old failed to show a robust late, right-prefrontal EM effect, which was again present in the ERPs of the young. By contrast, the older adults showed a central negative component not seen in the ERPs of the young. These results are consistent with the frontal lobe deficit hypothesis of aging and provide some evidence that old and young may use distinct cortical networks during source memory retrieval. PMID- 11919000 TI - The top down and bottom up mechanisms involved in the sudden awareness of low level sensorimotor behavior. AB - Motor control can be achieved in the absence of awareness, even when performed intentionally. The aim of this study was to understand the mechanisms of the sudden awareness of our own movement. This was studied in locomotion because it is an automatic behavior which can be intentionally modulated. Subjects walked continuously with the instruction to maintain either a constant walking speed (compensation condition) or constant propulsive forces (no-intervention condition); they were sometimes faced with slow variations in resistance that they had to detect. The results show that: (1) the subject remains unaware of his force increase (in compensation) or his walking velocity decrease (in no intervention) for a long time, although these modifications go largely beyond the variability range in which he is able to intentionally control his force (in no intervention) or his velocity (in compensation) and (2) the detection of the resistance increase occurs at the same time in both conditions. We conclude that the sudden awareness of a movement pattern produced at a low level was found to emerge from the interaction between a top down mechanism where the intentional control of goal feedback delays the aware perception of the other sensory sources and a bottom up mechanism where high level mechanisms of sensorimotor integration come into play beyond a discrepancy threshold between different sensory information. PMID- 11919001 TI - Event-related potentials during discourse-level semantic integration of complex pictures. AB - This study examined event-related potentials (ERPs) elicited in response to semantic processing of non-verbal stories. ERPs were recorded from 29 scalp electrodes on 16 participants while they viewed series of complex gray-scale pictures, each of which relayed a simple story. The final picture of each story was either congruous or incongruous with the preceding context. Participants made delayed meaningfulness judgments for each story. Averaged ERPs time-locked to the onset of the final picture were more negative for incongruous than congruous pictures. Two distinct components were sensitive to congruency. The first component peaked at approximately 325 ms (N300) and was distributed over central and frontal sites. The second component peaked at approximately 500 ms and also had a centro-frontal maximum but was more widespread than the earlier component (anterior N400). The distinct scalp topographies of these two negativities provide strong evidence that the N300 and N400 are separate and distinguishable components. Furthermore, the presence of the N300 in this exclusively pictorial task suggests that the N300 is specific to the semantic processing of non-verbal stimuli and is not due to linguistic mediation. This study also revealed that the N400 can be modulated by discourse-level coherence manipulations with pictures. Finally, the different patterns of ERP effects observed during the semantic processing of verbal and non-verbal information suggest that non-identical neuronal sources, and thus non-identical representational systems, are accessed by these different types of materials. These findings strongly support at least partial modularity of semantic representations and processing mechanisms in the human brain. PMID- 11919002 TI - Effects of picture repetition on induced gamma band responses, evoked potentials, and phase synchrony in the human EEG. AB - Repeated experience with an object due to prior exposure to that object is commonly referred to as perceptual or repetition priming. One possible neuronal mechanism for repetition priming is 'repetition suppression' within a cell assembly coding the stimulus. Recently, induced gamma band responses (GBRs) were discussed as a possible physiological correlate of activity in such a cell assembly. The present EEG study was designed to investigate the modulation of induced GBRs when line drawings were presented either once or consecutively two or three times. Results showed a broad distribution of spectral gamma power and synchrony after initial picture presentation. Repeated presentations of the same picture led to a decrease of induced gamma power and less synchronized activity between distant electrode sites. The decrease of induced GBRs and synchrony after repeated picture presentations may be linked to a 'neural savings' mechanism within a cell assembly representing an object. Furthermore, the visual evoked potential, which was modulated by priming, showed a topographically different distribution compared to induced GBRs. PMID- 11919003 TI - Top-down effects can modify the initially stimulus-driven auditory organization. AB - We recorded event-related potentials (ERPs) and magnetic fields (ERFs) of the human brain to determine whether top-down control could modulate the initial organization of sound representations in the auditory cortex. We presented identical sound stimulation and manipulated top-down processes by instructing participants to either ignore the sounds (Ignore condition), to detect pitch changes (Attend-pitch condition), or to detect violations of a repeating tone pattern (Attend-pattern condition). The ERP results obtained in the Attend pattern condition dramatically differed from those obtained with the other two task instructions. The magnetoencephalogram (MEG) findings were fully compatible, showing that the neural populations involved in detecting pattern violations differed from those involved in detecting pitch changes. The results demonstrate a top-down effect on the sound representation maintained in auditory cortex. PMID- 11919004 TI - Maximum cognitive performance and physiological time trend measurements after caffeine intake. AB - The effect of caffeine on the central nervous system and cardiorespiratory system was tested under resting conditions and while undertaking a multitask performance test. The subjects abstained from caffeine for a week before the study. Each subject performed the test after oral administration of 90 and 250 mg of caffeine on two separate days. Sum of Squares was recorded during the whole period of the study. Heart rate (HR) and respiration rates (RR) were continuously recorded and blood pressure (BP) was recorded before and after each stage of the experiment (Test(0), Test(1), Test(2), Test(3)). Sixteen healthy volunteers participated in the study divided into three groups: Group A, non-smokers and non-coffee drinkers; Group B, smokers and coffee drinkers; and Group C, non-smokers and coffee drinkers. Comparison of the performance of each stage with the resting conditions revealed statistically significant differences of group B compared to the other two groups and no significant differences between Groups A and C in both doses of caffeine. Non-coffee drinkers needed a low dose of caffeine for their optimal performance while a higher dose significantly increased their blood pressure. Coffee drinkers and smokers needed a higher dose of caffeine for optimal performance, which increased very quickly, but did not last and increased their BP. This increase in BP was not statistically significant, probably because of nicotine's effect. Heart rate was decreased and respiration rate increased significantly. The optimal performance was dose-dependent, increasing significantly with the higher dose of caffeine but with adverse effects on BP and RR. PMID- 11919005 TI - Processing of changes in visual speech in the human auditory cortex. AB - Seeing a talker's articulatory gestures may affect the observer's auditory speech percept. Observing congruent articulatory gestures may enhance the recognition of speech sounds [J. Acoust. Soc. Am. 26 (1954) 212], whereas observing incongruent gestures may change the auditory percept phonetically as occurs in the McGurk effect [Nature 264 (1976) 746]. For example, simultaneous acoustic /ba/ and visual /ga/ are usually heard as /da/. We studied cortical processing of occasional changes in audiovisual and visual speech stimuli with magnetoencephalography. In the audiovisual experiment congruent (acoustic /iti/, visual /iti/) and incongruent (acoustic /ipi/, visual /iti/) audiovisual stimuli, which were both perceived as /iti/, were presented among congruent /ipi/ (acoustic /ipi/, visual /ipi/) stimuli. In the visual experiment only the visual components of these stimuli were presented. A visual change both in audiovisual and visual experiments activated supratemporal auditory cortices bilaterally. The auditory cortex activation to a visual change occurred later in the visual than in the audiovisual experiment, suggesting that interaction between modalities accelerates the detection of visual change in speech. PMID- 11919006 TI - An event-related functional MRI study comparing interference effects in the Simon and Stroop tasks. AB - The Stroop and Simon tasks typify a class of interference effects in which the introduction of task-irrelevant stimulus characteristics robustly slows reaction times. Behavioral studies have not succeeded in determining whether the neural basis for the resolution of these interference effects during successful task performance is similar or different across tasks. Event-related functional magnetic resonance imaging (fMRI) studies were obtained in 10 healthy young adults during performance of the Stroop and Simon tasks. Activation during the Stroop task replicated findings from two earlier fMRI studies. These activations were remarkably similar to those observed during the Simon task, and included anterior cingulate, supplementary motor, visual association, inferior temporal, inferior parietal, inferior frontal, and dorsolateral prefrontal cortices, as well as the caudate nuclei. The time courses of activation were also similar across tasks. Resolution of interference effects in the Simon and Stroop tasks engage similar brain regions, and with a similar time course. Therefore, despite the widely differing stimulus characteristics employed by these tasks, the neural systems that subserve successful task performance are likely to be similar as well. PMID- 11919009 TI - Vaccine-associated feline sarcoma--an emerging problem. PMID- 11919011 TI - Current understanding of vaccination site-associated sarcomas in the cat. PMID- 11919010 TI - Cytokines and anti-cytokine therapy: clinical potential for treatment of feline disease. AB - Cytokines are soluble proteins produced by nucleated cells throughout the body. They have wide ranging effects on cell growth and differentiation, mediating immune responses, haemopoiesis and tissue repair. Advances in recombinant DNA technology have led to a vast increase in knowledge of their biological properties and subsequently their use in human clinical trials. The use of human cytokines in feline medicine has been of limited success as the action of cytokines is often species restricted or their activity may be neutralized due to antibody formation. Recently, however, many feline cytokines have been cloned which raises the possibility of their future use in the management and prevention of feline disease. Ultimately, they may find widespread clinical uses including the treatment of cancer, cytopenias and viral infections and as vaccine adjuvants. PMID- 11919012 TI - Disseminated Mycobacterium genavense infection in a FIV-positive cat. AB - An 8-year-old FIV-positive Australian cat was presented with coughing, periocular alopecia, pyrexia and inappetence. Skin scrapings demonstrated Demodex cati mites. Antibiotics were administered and it was treated successfully for periocular demodectic mange, but the cat continued to exhibit respiratory signs and lose weight. Further investigation revealed an ascarid infection and active chronic inflammation of undetected cause affecting the lower airways. Repetitive treatment with pyrantel failed to eradicate the ascarid infection. The cat became cachectic and developed moist ulcerative dermatitis of the neck, severe non regenerative anaemia, leucopenia and thrombocytopenia. Necropsy and histopathology revealed mycobacteriosis affecting skin, lungs, spleen, lymph nodes, liver and kidney. Attempted culture of frozen tissues at a mycobacteria reference laboratory was unsuccessful. Paraffin-embedded, formalin-fixed tissue was retrieved and examined using PCR to amplify part of the 16S rRNA gene. A diagnosis of disseminated Mycobacterium genavense infection was made based on the presence of acid fast bacteria in many tissues and partial sequence of the 16S rRNA gene. Although M genavense has been identified previously as a cause of disseminated disease in AIDS patients, this is the first report of infection in a cat. It was suspected that the demodecosis, recurrent ascarid infections and disseminated M genavense infection resulted from an immune deficiency syndrome consequent to longstanding FIV infection. PMID- 11919013 TI - The clinical efficacy of topical and systemic therapy for the treatment of feline ocular chlamydiosis. AB - Twenty-four specific-pathogen-free-derived cats aged four to 11 months were challenged by ocular application of a field isolate of Chlamydia psittaci to evaluate the effect of topical and systemic therapy on the course of disease. The cats were monitored for 35 days post-challenge, with severity of clinical signs being measured using a scoring system, and ocular shedding of the organism monitored by culture of conjunctival swabs. All cats developed active C psittaci infection, and after 7 days the cats were randomly assigned to one of four treatment groups: Group P (placebo) was given twice-daily ophthalmic tear replacement ointment; group F was given twice-daily topical 1% fusidic acid ophthalmic viscous drops; group C was given twice-daily topical 1% chlortetracycline ophthalmic ointment; and group D was given doxycycline at 10 mg/kg daily per os in addition to twice-daily topical 1% fusidic acid ophthalmic ointment. Within 24 h of commencement of therapy, group D had significantly lower median clinical scores than group P, and with the exception of day 16, this trend was maintained throughout the observation period. Median clinical scores of cats in group F were not appreciably different to those in group P, whereas the median scores of cats in group C generally fell between those of groups P and D. The median duration of C psittaci shedding was 10 and 15 days for groups D and C respectively, but four of the six cats in groups F and P were still shedding organisms at the end of the study (day 35). In this study, systemic therapy with doxycycline proved superior to topical therapy in the treatment of feline chlamydiosis. PMID- 11919014 TI - Efficacy of oral terbinafine in feline dermatophytosis due to Microsporum canis. AB - Microsporum canis is the dermatophyte most commonly responsible for ringworm in cats. The purpose of this paper was to evaluate the in vivo efficacy of oral terbinafine (Lamisil; Sandoz) in the treatment of feline ringworm caused by M canis, and to consider this drug as an alternative to griseofulvin or imidazoles. Fifteen cats infected with M canis were treated orally once daily with 30 mg/kg of terbinafine over a 2-week period. All treated animals were checked for dermatophytes on the last day of treatment, a month later and 3 months after the last administration of the drug. Only 12 cats could be used in the whole trial and 11 of these (92%) showed a complete cure. Terbinafine could be an effective alternative to griseofulvin when fungal resistance or idiosyncrasic intolerance are shown and, compared with griseofulvin, could give a faster rate of cure and less relapses. PMID- 11919015 TI - Intrathoracic tracheal rupture. AB - This paper reports five new cases of intrathoracic tracheal rupture in cats, and summarizes these cases in conjunction with 11 cases from the literature. Most cats had no obvious respiratory signs at the time of injury, and in half of them the tracheal rupture was the only injury. The interval from trauma to onset of dyspnoea ranged from 1 to 28 days (median 12.5 days). Radiographic findings include loss of continuity of the trachea, often with a gas-filled diverticulum present between separated tracheal rings. With careful attention to surgical technique and anaesthetic management the prognosis for these cats is excellent. All eight of the cats reported in the last 14 years having surgical correction survived and returned to normal. The surgical anatomy, approach and tracheal anastomosis technique is described and recommendations for anaesthetic management are made. PMID- 11919016 TI - Obstructive inflammatory laryngeal disease in three cats. AB - Three cats with upper respiratory tract obstruction due to laryngeal inflammation are presented. Cervical radiography showed the presence of a soft tissue mass in the laryngeal region in all cases, and laryngoscopy allowed direct visualization of a mass associated with the larynx. Laryngeal samples were obtained by a combination of fine needle aspiration, cutting biopsy forceps, by ventral laryngotomy, and at post-mortem. Histopathology of the laryngeal samples showed the presence of a predominantly granulomatous inflammation, with macrophage and lymphocyte infiltration. One case was euthanased due to severe dyspnoea. The remaining two cases underwent combined medical (corticosteroid and antibiotic) and surgical (permanent tracheostomy or excision of laryngeal tissue by ventral laryngotomy) treatment. One case died of an undetermined cause 15 weeks after surgery while the other case remains clinically well 20 months after diagnosis. Recognition of the existence of granulomatous laryngitis is important as clinical signs and radiographic findings are indistinguishable from laryngeal neoplasia. PMID- 11919017 TI - Congenital urethral anomaly in a kitten. PMID- 11919019 TI - PCR and feline medicine. PMID- 11919020 TI - Feline vestibular disorders. Part I: anatomy and clinical signs. AB - An understanding of the anatomy of the feline vestibular system is essential for interpretation of the clinical signs associated with vestibular dysfunction, for precise lesion localisation, and for accurate interpretation of results of diagnostic imaging. Appropriate recognition and interpretation of the clinical signs of vestibular disease is also an essential aspect of the precise diagnosis of the cause of vestibular dysfunction in cats. The objectives of this review are to provide an overview of the anatomy of the feline vestibular system, and to review the clinical signs of peripheral and central vestibular dysfunction of cats. PMID- 11919021 TI - Feline vestibular disorders. Part II: diagnostic approach and differential diagnosis. AB - Results of a neurological examination usually permit localisation of a vestibular disorder to either the central or peripheral parts of the vestibular system. Many different disorders located in the same part of the vestibular system will produce similar clinical signs. Therefore, additional diagnostic tests beyond a neurological examination are required in order to make an accurate diagnosis. The objectives of this review are to outline a diagnostic approach for disorders affecting the feline vestibular system, and to summarise the clinically important features of frequently diagnosed diseases affecting the vestibular system of cats. PMID- 11919022 TI - The role of polymerase chain reaction and its newer developments in feline medicine. AB - We give a brief overview on the principles of the polymerase chain reaction (PCR), reverse transcriptase PCR (RT-PCR), quantitative competitive PCR and real time PCR (TaqMan technology). The literature dealing with PCR and its role in the diagnosis, pathogenesis and research of infectious diseases of the domestic cat is reviewed. Cross-contaminations which occasionally occur during handling of amplified DNA may be an important problem in the PCR laboratory. In many infectious diseases, PCR results are difficult to interpret as their predictive positive and negative values are not always known. Newer assays, such as TaqMan procedures, are becoming increasingly reliable and cost-effective. It can be expected that additional knowledge on how to interpret PCR results will soon be available. PMID- 11919023 TI - Two cases of feline visceral and cutaneous leishmaniosis in Spain. AB - This paper describes clinical signs and lesions in two cases of leishmaniosis- one visceral and one cutaneous in the cat (Felis catus domesticus). The diagnosis was achieved by a combination of serology, light and electron microscopic studies. The vague nature of the clinical signs observed in both cases was particularly striking, and clinical features were similar to many other diseases commonly found in cats. Therefore, the use of various investigations to detect leishmaniosis (serum chemistry, serology and histopathology) is highly recommended in cases where clinical signs do not respond to conventional treatment. PMID- 11919024 TI - Insulin sensitivity in normal and diabetic cats. AB - Estimates of in vivo insulin sensitivity (S(I)) can be derived from minimal model analysis of a frequently sampled intravenous glucose tolerance test (FSIVGTT). Modification of the FSIVGTT by the injection of insulin allows insulin sensitivity to be measured in diabetics. To establish and compare reference values for insulin sensitivity in clinically normal and diabetic cats, we subjected 10 clinically normal cats and five diabetic cats to the insulin modified FSIVGTT with minimal model analysis. Diabetic cats had a significantly lower insulin sensitivity than clinically normal cats (P<0.05). Mean insulin sensitivity in clinically normal cats was 3.22x10(-4)/min/microU/ml (range 1.71 5.23x10(-4)/min/microU/ml). In contrast, the mean insulin sensitivity in diabetic cats was 0.58x10(-4)/min/microU/ml (range 0.136-0.88x10(-4)/min/microU/ml), or approximately six times less insulin sensitive than clinically normal cats. Mean glucose effectiveness in clinically normal cats was 0.030/min (range 0.021 0.045/min). Mean glucose effectiveness in diabetic cats was 0.014/min (range 0.008-0.021/min). Our data demonstrate that insulin resistance is a feature of feline diabetes mellitus and that diabetic cats have a similar relative decrease in insulin sensitivity to humans with type 2 diabetes. PMID- 11919026 TI - Suspected hypersensitivity to phenobarbital in a cat. PMID- 11919025 TI - Successful surgical treatment of a cat with primary aldosteronism. PMID- 11919027 TI - Surgical options for the treatment of hyperthyroidism in the cat. AB - Since the first description of feline hyperthyroidism in 1978, numerous treatment options for hyperthyroidism have been reported. Surgical removal of enlarged, autonomously functioning thyroid glands is one of the most commonly used treatment options. Affected cats must have a careful pre-operative evaluation to detect concurrent medical conditions such as renal disease or cardiomyopathy. Since more than 80% of hyperthyroid cats have neoplastic changes in both thyroid glands, bilateral thyroidectomy is necessary for treatment of the majority of hyperthyroid cats. Several different thyroidectomy techniques have been developed in an attempt to minimise potential post-operative complications associated with bilateral thyroidectomy such as hypocalcemia or recurrence of hyperthyroidism. Damage to or removal of all four parathyroid glands during bilateral thyroidectomy causes hypocalcemia, the most common post-operative complication. Recurrence of hyperthyroidism can occur months after initial thyroidectomy if residual adenomatous thyroid tissue is retained in the surgical site. The most effective surgical techniques for bilateral thyroidectomy involve preservation of at least one external parathyroid gland on the surface of the thyroid capsule. Additionally, the majority of the thyroid capsule must be removed to ensure that all neoplastic thyroid tissue is removed. The most recently described feline thyroidectomy techniques involve sequential removal of bilaterally affected thyroid glands. Staging a bilateral thyroidectomy allows time for ipsilateral parathyroid tissue to revascularise before the second thyroid gland is removed and the blood supply to the contralateral parathyroid glands is potentially interrupted. Thyroidectomy is a very effective treatment option for hyperthyroid cats. Surgical treatment of hyperthyroidism in cats offers permanent cure without chronic medical management. No specialised equipment other than standard surgical instrumentation and facilities are necessary. With practice, feline thyroidectomy can become a routine procedure in most veterinary hospitals. PMID- 11919028 TI - Hereditary and congenital ocular disease in the cat. AB - The aim of this review of hereditary and congenital ocular disease in cats is to present an overview of the most common disorders seen in this species, the pathogenesis of the problems and wherever possible, how they are treated. Several defects are common in breeds such as the Persian, Himalayan and Burmese cats and affect the anterior segment of the eye. Examples are agenesis of the eyelids, dermoids, entropion and corneal sequestrum. Other problems such as cataracts, lens luxation and retinal dysplasia, cause problems of the intraocular structures, but are less common in cats compared to dogs. Finally, various parts of the retina and in some diseases other parts of the eye, are specifically affected by hereditary diseases. Examples of these are lysosomal storage disease, Chediak-Higashi syndrome and progressive rod cone degeneration and rod cone dysplasia. Research of the latter two hereditary diseases, both described in the Abyssinian breed of cat, have made affected individuals important animal models for research into comparable diseases of humans. PMID- 11919029 TI - Current understanding of feline diabetes: part 1, pathogenesis. AB - Type-1 diabetes, resulting from immune-mediated destruction of beta cells, appears to be rare in cats. Type-2 diabetes, characterised by inadequate insulin secretion and impaired insulin action, is the most common form of diabetes in cats. Other specific forms of diabetes constitute a substantial minority of cases. The most common is pancreatic destruction from pancreatic adenocarcinoma. Less frequent causes are insulin resistance from other endocrinopathies including acromegaly. Diabetes in cats is characterised by variable loss of insulin secretory capacity and insulin resistance. Glucose toxicity, islet amyloid deposition, and pancreatitis contribute to further loss of beta cells and failure of insulin secretion. A significant number of cats undergo remission of their diabetes, usually 1-3 months after good glycaemic control is instituted. Obesity, old age, and Burmese breed are recognised risk factors for the development of diabetes in cats. PMID- 11919030 TI - Feline inflammatory bowel disease: a review. AB - Inflammatory bowel disease (IBD), while a popular diagnosis, may not occur as commonly as it is diagnosed. It is a diagnosis of exclusion, meaning that it is important to eliminate diseases that mimick it. Dietary intolerance or allergy in particular, can have the same clinical and histologic appearance as IBD. Likewise, well-differentiated alimentary lymphosarcoma can also be confused with it. Intestinal biopsies are useful, but must be taken carefully and then evaluated by someone with interest and expertise in alimentary tract pathology. Therefore, it behoves the clinician to carefully consider the diagnosis instead of starting multiple drug therapy in a cavalier fashion. Well constructed dietary therapy can often be beneficial for both dietary problems and IBD. PMID- 11919031 TI - Primary renal tumours in cats: 19 cases (1992-1998). AB - A search from databases of four veterinary colleges and one private referral practice between January 1992 and April 1998 provided 20 cases diagnosed with primary renal neoplasia. Review of these cases revealed 19 primary renal tumours, excluding lymphoma. Of the 20 histologically reviewed cases, the diagnosis was amended in eight. There were 13 renal carcinomas (11 tubular and two tubulopapillary), three transitional cell carcinomas, one malignant nephroblastoma, one haemangiosarcoma and one adenoma. The haemangiosarcoma is, to our knowledge, the first reported case of this tumour type as a primary renal tumour in the cat. Most cats were presented for non-specific clinical signs such as anorexia and weight loss. One cat presented with tumour-associated polycythaemia which has not, to our knowledge, been reported previously. The metastatic rate for cats with complete staging was 64%, and 100% for transitional cell carcinomas. PMID- 11919032 TI - Vegetative endocarditis in six cats. AB - Between 1990 and 1997 vegetative endocarditis was diagnosed in six neutered cats (three males, three females) aged between 3 and 9 years. Two cats were purebred (one Persian and one Tonkinese), the remaining four being domestic short haired cats. The diagnosis was made using echocardiography (five cases) or at necropsy (one case). Concurrent involvement of the aortic and mitral valve was noted in four cats, the aortic valve alone was affected in one case, and the tricuspid valve in another. A likely microbiological diagnosis was obtained in three cats: a Bartonella species in two cats based on positive blood culture and a Streptococcus species in one cat based on Gram stain of valve lesions at necropsy. In another cat, Gram-positive cocci were demonstrated histologically in tricuspid vegetations. Invariably, cats had signs of congestive heart failure (left-sided in five cats, right-sided in one), and this was the major factor contributing to mortality in four cases. Signs referable to sepsis were prominent in only two patients. Appropriate medical therapy, consisting of antimicrobials and drugs to treat congestive heart failure, resulted in survival for 5 and 11 months, respectively, in two cases. The other cats died within 2 weeks of diagnosis, including two which received aggressive treatment in hospital. PMID- 11919033 TI - Recommendations for the housing of cats in the home, in catteries and animal shelters, in laboratories and in veterinary surgeries. AB - In recent years, there has been an increase in interest in applied ethology and animal welfare, and an increase in the popularity of the domestic cat. This has stimulated research on the behaviour and welfare of cats kept in different environments. This article presents a review of the recent research and makes recommendations for the housing of domestic cats in the home, in catteries and animal shelters, in laboratories and in veterinary surgeries. PMID- 11919034 TI - Circulating lupus anticoagulant and probable systemic lupus erythematosus in a cat. AB - An adult domestic short hair cat was presented in a critical condition with icterus. Various investigations demonstrated the presence of haemolytic anaemia and hepatic abnormalities, as well as significant coagulation defects. Systemic lupus erythematosus (SLE) was suggested as a possible cause. PMID- 11919035 TI - The weathering hypothesis. PMID- 11919036 TI - Questionnaire wording on population-based estimates of mammography prevalence. PMID- 11919040 TI - Colorful communities: toward a language of inclusion. PMID- 11919041 TI - Elevating the voices of rural minority women. PMID- 11919042 TI - Moving forward: addressing the health of Asian American and Pacific Islander women. AB - Little is known about the health of Asian American and Pacific Islander (AAPI) women, a rapidly growing population marked by diverse sociodemographic characteristics, health needs, and access to and use of health services. This commentary provides broad recommendations for research, program development, and policy development based on the first-ever White House Initiative report on AAPIs. These recommendations address the issues of data, access, civil rights, community capacity, and the need to recognize ethnic subgroups among the AAPI population. Reflecting on the events of the past year, the recommendations provide direction for public health to address the health and well-being of AAPI women. PMID- 11919043 TI - Reconceptualizing native women's health: an "indigenist" stress-coping model. AB - This commentary presents an "indigenist" model of Native women's health, a stress coping paradigm that situates Native women's health within the larger context of their status as a colonized people. The model is grounded in empirical evidence that traumas such as the "soul wound" of historical and contemporary discrimination among Native women influence health and mental health outcomes. The preliminary model also incorporates cultural resilience, including as moderators identity, enculturation, spiritual coping, and traditional healing practices. Current epidemiological data on Native women's general health and mental health are reconsidered within the framework of this model. PMID- 11919044 TI - Public health needs and scientific opportunities in research on Latinas. AB - Much of the research on women's health has not deepened our understanding of health issues affecting Latinas. Yet integration of research on Latinas into the women's health agenda is important for at least 2 reasons. First, critical public health issues facing Latinas must be better understood if effective interventions designed to eliminate racial and ethnic disparities in health are to be developed and implemented. Second, studies on the health of Latinas represent unique opportunities to advance scientific understandings of underlying processes relevant to the health of other populations. Such research can further our knowledge of the processes underlying cultural adaptation and negotiation of changing sex roles and how these issues affect the health of women. Critical research and empirical approaches that help us to understand how race, ethnicity, sex, and class shape the health of Latinas will inform broader public health issues. PMID- 11919045 TI - Intimate partner violence and women of color: a call for innovations. AB - In this commentary, we focus on violence against women of color. Although African American women experience higher rates of intimate partner homicide than White women, the cumulative rates for nonfatal intimate partner violence are similar and do not vary between urban and rural locations (though access to services may vary by location). Much of the research about intimate partner violence is based on women with low socioeconomic status and on interventions that were developed by and for White women. Current primary prevention strategies focus on violence that is perpetrated by strangers rather than their primary perpetrators--intimate partners. We recommend the development and rigorous evaluation of prevention strategies that incorporate the views of women of color and attention to primary prevention. PMID- 11919046 TI - What factors hinder women of color from obtaining preventive health care? AB - This commentary examines how women of color fare on the use of preventive care. Logistic regression models of women's use of preventive care were computed with data from the 1994 Commonwealth Fund Minority Health Survey. It was found that having a regular doctor was the most consistent predictor of the use of preventive care, irrespective of the women's racial/ethnic background, socioeconomic circumstances, or place of residence. These findings reinforce the importance of physicians in the delivery of preventive care. Suggestions for improving the use of preventive services by women of color are provided. PMID- 11919047 TI - Impact of breast cancer on African American women: priority areas for research in the next decade. AB - Despite all the gains that have been made in the area of breast cancer research, African American women suffer disproportionately from the effects of the disease. Breast cancer is the second leading cause of cancer death among African American women, exceeded only by lung cancer. Improvements in cancer incidence, mortality, and survival rates among populations are undoubtedly the outcome of quality research. Therefore, there is a need to identify and discuss issues regarding breast cancer among African American women and to determine whether these issues should be a part of the nation's breast cancer research agenda. This commentary summarizes the results of the Summit Meeting Evaluating Research on Breast Cancer in African American Women, which was held September 8-11, 2000, in Washington, DC. Listed are priority areas and some of the questions that fueled this 2-day discussion among 130 participants, including health advocates, cancer survivors, and experts representing various areas of cancer research. PMID- 11919048 TI - Diabetes, diversity, and disparity: what do we do with the evidence? AB - The US Department of Health and Human Services has developed an initiative called "Eliminating Racial and Ethnic Disparities in Health," which parallels Healthy People 2010, the nation's health goals for the next decade. The initiative focuses on areas of health disparity that are known to affect racially and ethnically diverse groups of the population yet hold the promise of improvement. The first step to addressing such health inequities is to understand the scope and nature of the diseases that contribute to such disparities. This commentary reviews the epidemiology and consequences of type 2 diabetes, particularly as it is manifested in socially and culturally diverse groups, and offers recommendations for actions to address the disparities resulting from diabetes. PMID- 11919049 TI - Cardiovascular disease among women residing in rural America: epidemiology, explanations, and challenges. AB - Many believe that the United States has entered a "Golden Age" of cardiovascular health and medicine. Pharmacological and technological advances have indeed produced an era of declining mortality rates from cardiovascular diseases for the nation as a whole. However, there remain areas of challenge. Cardiovascular disease (CVD) is still by far the leading cause of death and disability in the United States, and it is the leading killer of US women. Perhaps the single most notable feature of the CVD epidemic in the United States is the substantial difference in morbidity and mortality that exists between White women and women of color, with a disproportionate share of suffering borne by minority women. Unexplained regional variations also cloud the otherwise notable progress of the last 30 years, and many rural areas appear to be uniquely affected by cardiovascular disease. This commentary reviews the evidence that the CVD epidemic disproportionately burdens women of color who reside in rural areas, itemizes and provides a logical framework for explaining this burden, and suggests approaches to solving this vexing public health problem. PMID- 11919050 TI - Toward a strategic approach for reducing disparities in infant mortality. AB - The United States' international ranking for infant mortality slipped from 19th in the 1980s to 27th in 1997. This slippage may be related to the segregation of priorities that occurred early in the 1990s, when national concern was diverted from infant mortality to minority health. To rekindle concern about infant mortality to the level of effective action, public health professionals must refocus the public's attention on assuring that all women are provided adequate education and services to help them avoid unintended pregnancies, that all pregnant women receive services in appropriate facilities, and that the causes of preterm deliveries are discovered. Effective action in these areas would not only improve infant mortality overall; it would also reduce racial and ethnic disparities in infant health. PMID- 11919051 TI - Spirituality and health for women of color. AB - Spirituality among African American and Hispanic women has been associated with a variety of positive health outcomes. The purposes of this commentary are (1) to define spirituality, comparing it with religiosity, and briefly examine the historical, cultural, and contextual roots of spirituality among women of color; (2) to explore research data that support a relationship between spirituality and health, particularly among women of color; and (3) to present several examples of how spirituality may enhance public health interventions designed to promote health and prevention. PMID- 11919052 TI - Considerations for community-based research with African American women. AB - Although community-based research is intended to be culturally sensitive, more advanced consideration of the impact of race, gender, and class is needed for health disparity research involving women of color. Research processes must permit the simultaneous disclosures of the racial, gender, and class identities among women of color that are assumed and imposed. The authoritative knowledge that women of color have about their lives and their health should form the basis for collaboration between researchers and study participants. The dissemination of research findings to study participants, and dialogue on those findings, is imperative for the development of sustainable interventions. PMID- 11919053 TI - A dedicated public health nurse. PMID- 11919054 TI - The face of women's health: Helen Rodriguez-Trias. AB - The American Public Health Association has announced that it will establish an award in the name of Helen Rodriguez-Trias, MD, its first Latina president, who died of lung cancer on December 27, 2001 [corrected]. Rodriguez-Trias, a nationally known advocate for underserved communities, was awarded the Presidential Citizens Medal by President Clinton in January 2001 for her work on behalf of children, women, people with AIDS, and the poor. This article is based on a dialogue with Rodriguez-Trias that began in September 2001 and ended December 12, 2001. PMID- 11919055 TI - Who does the work of public health? AB - Frequently, we take the "public" out of public health and allow the practice to become extremely narrow, limited to experts telling the public what's best for them. PMID- 11919056 TI - Mobilizing women for minority health and social justice in california. AB - Women's Health Leadership is building the leadership capacity of diverse community leaders in California committed to promoting health and social justice in their communities. This program provides opportunities for cross-cultural exchanges of ideas, resources, and expertise. Graduates continue to receive technical support and to engage in peer learning via an alumnae network. The network is dedicated to advancing social justice issues and to addressing health disparities. It is also a way to mobilize trained grassroots leaders to provide expertise to influence policy decisions, to provide technical support and resources to local communities, and to foster new partnerships across the state. PMID- 11919057 TI - Eliminating health disparities among minority women: a report on conference workshop process and outcomes. AB - A national conference convened in May 2001 explored health disparities among minority women. It included 5 one-hour workshops that randomly assigned each participant to 1 of 4 groups. Groups generated recommendations on conference topics and from these identified priority recommendations. Trained facilitators guided groups through brainstorming and weighted voting processes; individual recommendations were submitted in writing. Participants generated 598 recommendations, 71 of them voted as priorities; these were analyzed to capture participants' "messages." Central themes focused on access issues and cultural incompetence as deterrents to the elimination of health disparities and on education, funding, and community-based, community-driven research as mechanisms for change. Strategies for change included reinventing or expanding the role of minority communities and changing health care itself and "how" it does its work. The essential element in all recommendations was community leadership and control. PMID- 11919058 TI - Racial/ethnic variations in women's health: the social embeddedness of health. AB - This article provides an overview of the magnitude of and trends in racial/ethnic disparities in health for women in the United States. It emphasizes the importance of attending to diversity in the health profiles and populations of minority women. Socioeconomic status is a central determinant of racial/ethnic disparities in health, but several other factors, including medical care, geographic location, migration and acculturation, racism, and exposure to stress and resources also play a role. There is a need for renewed attention to monitoring, understanding, and actively seeking to eliminate racial/ethnic disparities in health. PMID- 11919059 TI - Safety-net institutions buffer the impact of Medicaid managed care: a multi method assessment in a rural state. AB - OBJECTIVES: This project used a long-term, multi-method approach to study the impact of Medicaid managed care. METHODS: Survey techniques measured impacts on individuals, and ethnographic methods assessed effects on safety-net providers in New Mexico. RESULTS: After the first year of Medicaid managed care, uninsured adults reported less access and use (odds ratio [OR] = 0.46; 95% confidence interval [CI] = 0.34, 0.64) and worse barriers to care (OR = 6.60; 95% CI = 3.95, 11.54) than adults in other insurance categories. Medicaid children experienced greater access and use (OR = 2.11; 95% CI = 1.21, 3.72) and greater communication and satisfaction (OR = 3.64; 95% CI = 1.13, 12.54) than children in other insurance categories; uninsured children encountered greater barriers to care (OR = 6.29; 95% CI = 1.58, 42.21). There were no consistent changes in the major outcome variables over the period of transition to Medicaid managed care. Safety net institutions experienced marked increases in workload and financial stress, especially in rural areas. Availability of mental health services declined sharply. Providers worked to buffer the impact of Medicaid managed care for patients. CONCLUSIONS: In its first year, Medicaid managed care exerted major effects on safety-net providers but relatively few measurable effects on individuals. This reform did not address the problems of the uninsured. PMID- 11919060 TI - Is breast cancer a disease of affluence, poverty, or both? The case of African American women. PMID- 11919061 TI - Effectiveness of media strategies to increase enrollment and diversity in the Women's Health Registry. PMID- 11919063 TI - Relation between racial discrimination, social class, and health among ethnic minority groups. AB - OBJECTIVES: This study explored associations between racism, social class, and health among ethnic minority people in England and Wales. METHODS: We conducted a series of regression analyses on cross-sectional data from the Fourth National Survey of Ethnic Minorities to explore the relation between different indicators of racism and health and household occupational class. RESULTS: Marked independent associations existed between reported experience of racism and perceptions of Britain as a "racist society," household social class, age, sex, and various mental and physical health indicators. These associations showed reasonable consistency across the different ethnic groups. CONCLUSIONS: The different ways in which racism may manifest itself (as interpersonal violence, institutional discrimination, or socioeconomic disadvantage) all have independent detrimental effects on health, regardless of the health indicator used. PMID- 11919065 TI - Early adult characteristics and mortality among inner-city African American women. AB - OBJECTIVES: This study examined predictors of longevity in a cohort of inner-city African American women. METHODS: Data were derived from a cohort study of inner city African American mothers whose median age in 1966 was 31 years. Analyses involved single-decrement life tables and pooled logistic regression. RESULTS: Giving birth for the first time before age 25 and having at least a high school education predicted longevity in this sample. Effects of later age at first delivery in terms of mortality risk were stronger after 55 years and, especially, after 70 years. CONCLUSIONS: The findings offer support for Geronimus's weathering hypothesis. Predictors of longevity among African Americans may be distinct from predictors for the population as a whole. PMID- 11919062 TI - A multilevel analysis of the relationship between institutional and individual racial discrimination and health status. AB - OBJECTIVES: This study examined whether individual (self-perceived) and institutional (segregation and redlining) racial discrimination was associated with poor health status among members of an ethnic group. METHODS: Adult respondents (n = 1503) in the cross-sectional Chinese American Psychiatric Epidemiologic Study were geocoded to the 1990 census and the 1995 Home Mortgage Disclosure Act database. Hierarchical linear modeling assessed the relationship between discrimination and scores on the Medical Outcomes Study Short-Form 36 and revised Symptom Checklist 90 health status measures. RESULTS: Individual and institutional measures of racial discrimination were associated with health status after control for acculturation, sex, age, social support, income, health insurance, employment status, education, neighborhood poverty, and housing value. CONCLUSIONS: The data support the hypothesis that discrimination at multiple levels influences the health of minority group members. PMID- 11919064 TI - Heterogeneity of health disparities among African American, Hispanic, and Asian American women: unrecognized influences of sexual orientation. AB - OBJECTIVES: This study compared health indicators among self-identified lesbians/bisexual women and heterosexual women residing in Los Angeles County. METHODS: Respondents were English-speaking Hispanic, African American, and Asian American women. Health status, behavioral risks, access barriers, and indicators of health care were assessed. RESULTS: Prevalence rates of chronic health conditions were similar among women in the 3 racial/ethnic groups. However, lesbians and bisexual women evidenced higher behavioral risks and lower rates of preventive care than heterosexual women. CONCLUSIONS: Among racial/ethnic minority women, minority sexual orientation is associated with increased health risks. The effects of sexual minority status need to be considered in addressing health disparities affecting this population. PMID- 11919066 TI - Increasing use of mammography among older, rural African American women: results from a community trial. AB - OBJECTIVES: A community trial was undertaken to evaluate the effectiveness of the North Carolina Breast Cancer Screening Program, a lay health advisor network intervention intended to increase screening among rural African American women 50 years and older. METHODS: A stratified random sample of 801 African American women completed baseline (1993-1994) and follow-up (1996-1997) surveys. The primary outcome was self-reported mammography use in the previous 2 years. RESULTS: The intervention was associated with an overall 6 percentage point increase (95% confidence interval [CI] = -1, 14) in community-wide mammography use. Low-income women in intervention counties showed an 11 percentage point increase (95% CI = 2, 21) in use above that exhibited by low-income women in comparison counties. Adjustment for potentially confounding characteristics did not change the results. CONCLUSIONS: A lay health advisor intervention appears to be an effective public health approach to increasing use of screening mammography among low-income, rural populations. PMID- 11919067 TI - HIV-associated histories, perceptions, and practices among low-income African American women: does rural residence matter? AB - OBJECTIVES: This study compared HIV-associated sexual health history, risk perceptions, and sexual risk behaviors of low-income rural and nonrural African American women. METHODS: A cross-sectional statewide survey of African American women (n = 571) attending federally funded Special Supplemental Nutrition Program for Women, Infants, and Children clinics was conducted. RESULTS: Adjusted analyses indicated that rural women were more likely to report not being counseled about HIV during pregnancy (P =.001), that a sex partner had not been tested for HIV (P =.005), no preferred method of prevention because they did not worry about sexually transmitted diseases (P =.02), not using condoms (P =.009), and a belief that their partner was HIV negative, despite lack of testing (P =.04). CONCLUSIONS: This study provided initial evidence that low-income rural African American women are an important population for HIV prevention programs. PMID- 11919068 TI - Does a history of trauma contribute to HIV risk for women of color? Implications for prevention and policy. AB - OBJECTIVES: We investigated history of abuse and other HIV-related risk factors in a community sample of 490 HIV-positive and HIV-negative African American, European American, and Latina women. METHODS: Baseline interviews were analyzed, and logistic regressions were used to identify predictors of risk for positive HIV serostatus overall and by racial/ethnic group. RESULTS: Race/ethnicity was not an independent predictor of HIV-related risk, and few racial/ethnic differences in risk factors for HIV were seen. Regardless of race/ethnicity, HIV positive women had more sexual partners, more sexually transmitted diseases, and more severe histories of abuse than did HIV-negative women. Trauma history was a general risk factor for women, irrespective of race/ethnicity. CONCLUSIONS: Limited material resources, exposure to violence, and high-risk sexual behaviors were the best predictors of HIV risk. PMID- 11919069 TI - Papanicolaou test use among reproductive-age women at high risk for cervical cancer: analyses of the 1995 National Survey of Family Growth. AB - OBJECTIVES: This study assessed the relationship between risk factors for cervical cancer and Papanicolaou (Pap) test use within the past year among reproductive-age women. METHODS: The 1995 National Survey of Family Growth, a demographic and reproductive health survey of 10 847 women aged 15 to 44, was analyzed with multiple logistic regression. RESULTS: Of the women, 62% reported having had a Pap test within the past year. Use was significantly higher among women with risk factors and among African American women. Use was significantly lower among uninsured, poor, and foreign-born women and among women with lower educational attainment and of "other" race/ethnicity. CONCLUSIONS: Strategies to improve Pap test use include (1) educational campaigns that inform women of cervical cancer risk factors and encourage screening and (2) increased support for programs that expand access to Pap tests. PMID- 11919071 TI - DNA oxidation or apoptosis: will the real culprit of dna damage in hyperoxic lung injury please stand up? PMID- 11919070 TI - Hepatitis C virus infection in young, low-income women: the role of sexually transmitted infection as a potential cofactor for HCV infection. AB - OBJECTIVES: We evaluated risk for hepatitis C virus (HCV) infection in women residing in low-income neighborhoods of northern California. METHODS: A population-based sample of 1707 women, aged 18 to 29, were surveyed and screened for sexually transmitted infections and HCV. RESULTS: Women infected with HCV (2.5%) were more likely to have a history of injection and noninjection drug use, to exchange sex for money or drugs, and to have sexually transmitted infections. HCV was independently associated with history of injection drug use, herpes simplex virus type 2 (HSV-2) infection, and heroin and cocaine use. CONCLUSIONS: Injection drug use is the highest risk exposure for HCV, but HSV-2 and noninjection drug use contribute significantly to increased risk. HCV prevention programs in impoverished areas should integrate drug treatment and sexually transmitted infection control. PMID- 11919072 TI - What is eotaxin doing in the pleura? Insights into innate immunity from pleural mesothelial cells. PMID- 11919073 TI - Regulation of eosinophil viability by cytokines. PMID- 11919074 TI - Blocking neutrophil influx reduces DNA damage in hyperoxia-exposed newborn rat lung. AB - Hyperoxia-induced neutrophil infux in neonatal rats may contribute to impaired lung development through oxidative DNA damage. To determine whether blocking neutrophil influx prevents DNA damage, we treated newborn rats with 95% O2 beginning at birth, and at 3 and 4 d with nonimmune immunoglobulin G (IgG) (control) or anti-cytokine-induced neutrophil chemoattractant (CINC). At 8 d, lungs were inflation-fixed. Random sections were labeled using terminal transferase nick end-labeling (TUNEL), and DNA oxidation was measured using anti 8-OH-2'-deoxyguanosine (OHdG). To determine whether hyperoxia-induced TUNEL represented apoptosis, we labeled sections with anti-Bax (proapoptotic) and anti Bcl-2 (antiapoptotic). We labled additional sections with anti-M30, directed against an epitope formed by caspase 6 digestion of cytokeratin 18 during apoptosis. Hyperoxia induced marked increases in TUNEL and OHdG signal in lung parenchymal cells, which was substantially prevented by treatment with anti-CINC. The large effects of hyperoxia on TUNEL were not accompanied by substantial effects on Bax, Bcl-2, or M30. We conclude that neutrophil influx during hyperoxia damages DNA by nicking and oxidation, and that blocking neutrophil influx can prevent this. Effects of 95% O2 on TUNEL are not primarily due to apoptosis in this model. Neutrophil-mediated oxidative DNA damage may contribute to abnormal lung development in newborns subjected to significant oxidative stress. PMID- 11919075 TI - Production of eosinophilic chemokines by normal pleural mesothelial cells. AB - Eosinophilic pleural effusion occurs in many diseases. The mechanisms of eosinophil accumulation are not well understood. We showed previously that eotaxin was readily detectable in most pleural effusions, and its concentration significantly correlated with eosinophil number. To test the hypothesis that pleural eotaxin is produced by resident mesothelial cells, we examined its production by normal pleural mesothelial cells (NPMC). Eotaxin was induced by tumor necrosis factor (TNF)-alpha or interleukin (IL)-4 and was drastically increased by their combination. In contrast, interferon (IFN)-gamma inhibited eotaxin production. Regulated on activation, normal T cells expressed and secreted (RANTES) was also induced by TNF-alpha and was drastically increased by the addition of IFN-gamma. These effects were observed at both protein and mRNA levels. Stabilization of RANTES mRNA was observed with IFN-gamma but not IL-4; neither cytokine stabilized eotaxin mRNA. Eosinophil chemoattractant activity in culture supernatants of NPMC stimulated with TNF-alpha plus IL-4 was diminished by an anti-eotaxin antibody; that induced by TNF-alpha plus IFN-gamma was attenuated by an anti-RANTES antibody. Thus, NPMC can produce eotaxin, and different cytokines act on NPMC to induce different chemokines by different mechanisms. IFN-gamma, a Th1 cytokine, acts at least at the posttranscriptional level to induce RANTES production, but it inhibits eotaxin production. In contrast, IL-4, a Th2 cytokine, acts at the transcriptional level to induce eotaxin. PMID- 11919076 TI - Interleukin-15 inhibits spontaneous apoptosis in human eosinophils via autocrine production of granulocyte macrophage-colony stimulating factor and nuclear factor kappaB activation. AB - Prolonged eosinophil survival, i.e., reduced apoptosis, is implicated in the pathogenesis of chronic allergic inflammation. Here we demonstrate that interleukin (IL)-15, in the presence or absence of tumor necrosis factor (TNF) alpha, reduces spontaneous apoptosis in freshly isolated human eosinophils. The prosurvival effect of IL-15 was abrogated by neutralizing antibody to granulocyte macrophage-colony stimulating factor (GM-CSF), although GM-CSF was not detected in conditioned media by ELISA. Additionally, the effect of IL-15 on spontaneous eosinophil apoptosis appeared to require nuclear factor-kappaB (NF-kappaB) activation based on evidence for NF-kappaB nuclear translocation and abrogation of the effect by the NF-kappaB inhibitor, Bay 11- 7082. Finally, the data demonstrate that IL-15 expression is higher in the submucosa of endobronchial tissues from subjects with moderate to severe asthma when compared with control subjects. Thus, our results suggest that IL-15, either alone or in combination with TNF-alpha, may perpetuate allergic inflammation by reduction of spontaneous eosinophil apoptosis through autocrine production of GM-CSF and NF-kappaB activation. PMID- 11919077 TI - Deficiency in inducible nitric oxide synthase protects mice from ozone-induced lung inflammation and tissue injury. AB - Inhalation of ozone causes Type I epithelial cell necrosis and Type II cell hyperplasia and proliferation. This is associated with an accumulation of activated macrophages in the lower lung, which we have demonstrated contribute to tissue injury. Nitric oxide (NO) is a highly reactive cytotoxic macrophage derived mediator that has been implicated in lung damage. In the present studies we used knockout mice with a targeted disruption of the gene for inducible nitric oxide synthase (NOSII) to analyze the role of NO in ozone-induced lung inflammation and tissue injury. Treatment of wild-type control mice with ozone (0.8 ppm) for 3 h resulted in a time-dependent increase in protein and cells in bronchoalveolar lavage fluid, which reached a maximum 24-48 h after exposure. Alveolar macrophages isolated from animals treated with ozone were found to produce increased amounts of NO, as well as peroxynitrite. This was correlated with induction of NOSII protein and nitrotyrosine staining of lung macrophages in tissue sections and in culture. Production of superoxide anion and prostaglandin (PG)E2 by alveolar macrophages was also increased after ozone inhalation. In contrast, alveolar macrophages from NOSII knockout mice did not produce reactive nitrogen intermediates even after ozone inhalation. Moreover, production of PGE2 was at control levels. NOSII knockout mice were also protected from ozone-induced inflammation and tissue injury, as measured by bronchoalveolar lavage protein and cell number. There was also no evidence of peroxynitrite-mediated lung damage in these animals. Taken together, these data demonstrate that NO, produced via NOSII, and potentially, its reactive oxidative product peroxynitrite, play a critical role in ozone-induced release of inflammatory mediators and in tissue injury. PMID- 11919078 TI - Development of neural tissue and airway smooth muscle in fetal mouse lung explants: a role for glial-derived neurotrophic factor in lung innervation. AB - We have characterized the distribution of neural tissue and its primary target tissue, airway smooth muscle (ASM), in an in vitro mouse model of early lung development comprising left lung lobes at embryonic Day 12, cultured for 2 or 5 d. Neural tissue was detected with antibodies to protein gene product 9.5 (PGP 9.5), synapsin, and p75NTR (the low-affinity neurotrophin receptor), and smooth muscle with an antibody to alpha-actin. Imaging by confocal microscopy revealed few PGP 9.5-positive neurons at the start of culture; after 2 d clusters of neurons and nerve fibers had appeared along the lobar bronchus and after 5 d along the secondary and tertiary branches. Neural tissue did not just follow the smooth muscle-covered tubules, as seen in vivo, but also grew outside the lobes onto a wide layer of alpha-actin-positive cells, suggesting that smooth muscle may express a trophic factor that attracts nerves. Explants cultured with glial derived neurotrophic factor (GDNF) exhibited a striking increase in the amount of p75NTR- and PGP 9.5-positive tissue outside the lobes, whereas GDNF-impregnated beads attracted neuronal precursors and influenced the direction of neurite extension. We show that the mouse lung explant is suitable for investigating trophic signals involved in pulmonary innervation and that GDNF may have a role in the early innervation of the developing airways. PMID- 11919079 TI - Dose-related protection from nickel-induced lung injury in transgenic mice expressing human transforming growth factor-alpha. AB - To determine the role of transforming growth factor-alpha (TGF-alpha) in protecting the lung from aerosolized nickel injury, transgenic mouse lines expressing human TGF-alpha in the pulmonary epithelium, under control of the human surfactant protein-C gene promoter, were tested. Higher expressing TGF alpha transgenic mouse lines, expressing distinct levels of TGF-alpha, survived longer than nontransgenic control mice. Increased survival correlated with levels of TGF-alpha expression in the lung. After 72 h of nickel exposure (70 microg Ni/m3), transgenic lines with intermediate levels of the TGF-alpha expression demonstrated attenuation of lung injury. The highest expressing line (line 28) demonstrated reduced lung inflammation and edema, reduced lung wet-to-dry weight ratios, decreased bronchoalveolar lavage (BAL) protein and neutrophils, reduced interleukin (IL)-1beta, interleukin-6, and macrophage inflammatory protein-2, and maintained surfactant protein-B (SP-B) levels compared with nontransgenic controls. In the TGF-alpha transgenic mouse model, TGF-alpha protects against nickel-induced acute lung injury, at least in part, by attenuating the inflammatory response, reducing pulmonary edema, and preserving levels of SP-B. PMID- 11919080 TI - Protein tyrosine nitration in the ventilatory muscles: role of nitric oxide synthases. AB - Modification of tyrosine residues and formation of 3-nitrotyrosine is one of the most commonly identified effects of reactive nitrogen species on proteins. In this study we evaluated the presence and localization of tyrosine nitration in various ventilatory and limb muscles. We also assessed the contribution of the neuronal (nNOS), the endothelial (eNOS), and the inducible (iNOS) isoforms of nitric oxide synthase (NOS) to tyrosine nitration in skeletal muscles both under normal conditions and in response to severe sepsis. In normal rats and mice, muscle tyrosine nitration was detected at 52, 48, 40, 30, 18, and 10 kD protein bands. Tyrosine nitration of the majority of these protein bands was significantly reduced within 1 h of in vivo NOS inhibition in rats. Diaphragmatic protein tyrosine nitration in mice deficient in the inducible NOS (iNOS-/-) averaged ~ 50% of that detected in wild-type (iNOS+/+) mice. Injection of bacterial lipopolysaccharides (LPS) in rats produced a significant rise in protein tyrosine nitration in the mitochondrial and membrane fractions but not in the cytosol of ventilatory muscles. Absence of iNOS expression (iNOS-/-), but not nNOS (nNOS-/-) or eNOS (eNOS-/-), in genetically altered mice resulted in a significant reduction in LPS-mediated rise in diaphragmatic nitrotyrosine. We conclude that tyrosine nitration of proteins occurs in normal muscle fibers and is dependent mainly on the activity of the iNOS isoform. Sepsis-mediated increase in protein tyrosine nitration is limited to the mitochondria and cell membrane and is highly dependent on the activity of the iNOS but not the nNOS or eNOS isoforms. PMID- 11919081 TI - Neutrophil elastase induces MUC5AC gene expression in airway epithelium via a pathway involving reactive oxygen species. AB - Neutrophil-predominant airway inflammation and mucus obstruction of the airways are major pathologic features of chronic airway diseases, including cystic fibrosis and chronic bronchitis. Neutrophils release elastase, a serine protease that impairs mucociliary clearance and stimulates goblet cell metaplasia and mucin production. We previously reported that neutrophil elastase increases expression of a major respiratory mucin gene, MUC5AC, by enhancing mRNA stability. However, the molecular mechanisms of elastase-regulated MUC5AC expression are not known. We hypothesized that reactive oxygen species, generated by elastase treatment, mediate MUC5AC gene expression. To test this hypothesis, A549, a respiratory epithelial cell line, was treated with elastase in the presence or absence of the oxygen radical scavenger, dimethylthiourea, or the iron chelator, desferrioxamine. MUC5AC mRNA levels were assessed by Northern analysis. Both antioxidants significantly inhibited elastase-induced MUC5AC gene expression. Dimethylthiourea also inhibited the neutrophil elastase (NE)-induced increase in MUC5AC expression in normal human bronchial epithelial cells. To determine whether elastase treatment generated reactive oxygen species, A549 and normal human bronchial epithelial cells were loaded with dichlorodihydrofluorescein, a fluorescent indicator of oxidative stress. NE treatment increased cellular fluorescence in both cell types, indicating generation of intracellular reactive oxygen species. We conclude that NE treatment increases MUC5AC gene expression by an oxidant-dependent mechanism. PMID- 11919082 TI - Shear stress-mediated cytoskeletal remodeling and cortactin translocation in pulmonary endothelial cells. AB - Hemodynamic forces in the form of shear stress (SS) and mechanical strain imposed by circulating blood are recognized factors involved in the control of systemic endothelial cell (EC) cytoskeletal structure and function. However, the effects of acute SS on pulmonary endothelium have not been precisely characterized, nor the mechanism of rapid SS-induced EC cytoskeletal rearrangement understood. We exposed bovine and human pulmonary EC monolayers to laminar SS (10 dynes/cm2) in a parallel plate flow chamber and observed increased actin stress fiber formation 15 min after application of flow. Acute SS-induced pronounced cortical cytoskeletal rearrangement characterized by myosin light chain kinase (MLCK)- and Rho-associated kinase (RhoK)-dependent accumulation of diphosphorylated regulatory myosin light chains (MLC) in the cortical actin ring, junctional protein tyrosine phosphorylation, and transient peripheral translocation of cortactin, an actin-binding protein involved in the regulation of actin polymerization. SS-induced cortactin translocation was independent of Erk-1,2 MAP kinase, p60(Src), MLCK, or RhoK activities, and unaffected by overexpression of a cortactin mutant lacking four major p60(Src) phosphorylation sites. However, both SS-induced transient cortactin translocation and cytoskeletal reorientation in response to sustained (24 h) SS was abolished in cells overexpressing either dominant negative Rac 1 or a dominant negative construct of its downstream target, p21-activated kinase (PAK)-1. Our results suggest a potential role for cortactin in the SS-induced EC cortical cytoskeletal remodeling and demonstrate a novel mechanism of Rac GTPase-dependent regulation of the pulmonary endothelial cytoskeleton by SS. PMID- 11919083 TI - Tumor necrosis factor-alpha-induced secretion of RANTES and interleukin-6 from human airway smooth muscle cells: modulation by glucocorticoids and beta agonists. AB - Recent studies have demonstrated that tumor necrosis factor (TNF)-alpha stimulates the secretion of interleukin (IL)-6 and regulated on activation, normal T cells expressed and secreted (RANTES) from airway smooth muscle (ASM) cells, with the induction of each molecule being differentially regulated (IL-6 increased, RANTES inhibited) by cyclic adenosine monophosphate (cAMP)-elevating agents. In this study we identify the mechanisms mediating IL-6 and RANTES gene transcription in human ASM cells. We found that TNF-alpha induced IL-6 gene expression in ASM cells via a nuclear factor (NF)-kappaB-dependent pathway, whereas RANTES gene expression was mediated via activation of activator protein (AP)-1 and nuclear factor of activated T cells (NF-AT). TNF-alpha-induced IL-6 secretion was only partially inhibited by dexamethasone, yet TNF-alpha-induced RANTES secretion was abolished. beta-Agonists induced IL-6 secretion from ASM via activation of the CRE region of the IL-6 promoter. beta-Agonists augmented TNF alpha-induced IL-6 secretion, reflecting an additive effect of NF-kappaB and CRE response elements on IL-6 gene expression. In contrast, beta-agonists inhibited TNF-alpha-induced RANTES secretion via an AP-1-independent pathway. Collectively, these data elucidate transcriptional mechanisms mediating TNF-alpha-induced IL-6 and RANTES secretion from ASM cells, and identify the specific cis- or trans acting elements that determine the differential effects of glucocorticoids and cAMP-elevating agents on the expression of these genes. PMID- 11919084 TI - Cloning and functional analysis of the mouse 5-lipoxygenase promoter. AB - 5-lipoxygenase (ALOX5), an enzyme essential for the formation of all leukotrienes, is highly regulated at multiple levels, including gene transcription. The human ALOX5 promoter sequence has been cloned and is well characterized. Several important cis-acting elements have been identified including a G+C-rich sequence approximately 145-179 base pairs (bp) upstream from the ATG start codon. This region contains consensus-binding sites for the transcription factor serum protein 1, a zinc-finger transcription factor (SP1) and early growth-response protein 1, a zinc-finger transcription factor (EGR-1) and is unique in that functionally significant polymorphisms alter these sequences. To further understand the significance of these polymorphisms and other regulatory sequences in the promoter we cloned approximately 2,000 bp of the mouse promoter sequence from a 129/SvJ BAC library for direct comparison with the human gene. Like the human promoter, the mouse Alox5 promoter lacks a TATA box and has multiple start sites. The first 292 bp immediately upstream of the translational start site function as a core promoter that is capable of mediating high basal transcription in RAW cells but not 3T3 cells. There are vast differences in the distribution of consensus cis elements between human and mouse genes; however, three areas of strong homology exist and they contain consensus binding sites for the SP1, GATA, GGAGA, and ETS family of transcription factors. We show that Sp1/Sp3 is essential for constitutive promoter-reporter activity. PMID- 11919085 TI - Interleukin-4- and interleukin-13-enhanced transforming growth factor-beta2 production in cultured human bronchial epithelial cells is attenuated by interferon-gamma. AB - Cytokines derived from lymphocytes are believed to play key roles in a variety of diseases, including airway diseases such as asthma. The current study was designed to evaluate the hypothesis that cytokines derived from Th2 cells, interleukin (IL)-4 and IL-13, might contribute to tissue remodeling by modulating the production of transforming growth factor (TGF)-beta. In addition, the ability of interferon (IFN)-gamma, a cytokine derived from Th1 cells that can antagonize many effects of IL-4 and IL-13, was also assessed for its effects on TGF-beta production. IL-4 and IL-13 both stimulated production of TGF-beta2 release from human bronchial epithelial cells in a time- and concentration-dependent manner. Both with and without acidification, TGF-beta2 were detected. Neither TGF-beta1 nor TGF-beta3 was released. In contrast to the stimulatory effect on human bronchial epithelial cells, neither IL-4 nor IL-13 stimulated release of any TGF beta isoform from human lung fibroblasts. IFN-gamma reduced both basal, IL-4-, and IL-13-stimulated release of TGF-beta2 in human bronchial epithelial cells. The stimulatory effects of IL-4 and IL-13 and the inhibitory effect of IFN-gamma on TGF-beta2 release were paralleled by mRNA levels, as assessed by real-time reverse transcriptase-polymerase chain reaction (RT-PCR). In summary, the Th2 derived cytokines, IL-4 and IL-13, can stimulate production of TGF-beta from airway epithelial cells but not from lung fibroblasts. IFN-gamma, in contrast, can inhibit TGF-beta2 release both under basal conditions and following IL-4 or IL-13 stimulation. The ability of these cytokines to modulate TGF-beta release may contribute to both normal airway repair and to the development of subepithelial fibrosis in asthma. PMID- 11919086 TI - Regulation of capacitative and noncapacitative receptor-operated Ca2+ entry by rho-kinase in tracheal smooth muscle. AB - To determine the mechanisms of Ca2+ mobilization induced by receptor agonists, we examined the role of Rho-kinase on the sarcoplasmic reticulum (SR) Ca2+ stores dependent and -independent Ca2+ influx in guinea pig tracheal smooth muscle (TSM). Isometric tension and intracellular Ca2+ concentration ([Ca2+]i) were simultaneously measured using fura-2-loaded tissues. Depletion of the SR Ca2+ stores by thapsigargin caused an increase in [Ca2+]i and contraction, demonstrating capacitative Ca2+ entry (CCE). Because CCE was not inhibited by nifedipine, voltage-operated Ca2+ channels are not involved in CCE. Under the condition that CCE is fully activated, methacholine (MCh) and histamine caused further increases in [Ca2+]i and tension, demonstrating noncapacitative receptor operated Ca2+ entry (non-CCE). The Ca2+ influx and contraction via non-CCE was inhibited by Y-27632, a Rho-kinase inhibitor, in a concentration-dependent fashion. In contrast, Y-27632 did not affect thapsigargin-induced CCE. Cytochalasin D, which disrupts actin cytoskeleton, inhibited contraction induced by CCE or MCh with no change in [Ca2+]i. Our results indicate that not only CCE but also non-CCE exist in TSM and that the latter is regulated by Rho-kinase, independent of actin cytoskeleton. In conclusion, Ca2+ influx regulated by the RhoA/Rho-kinase pathway may play a functional role in contraction by agonists. PMID- 11919087 TI - Lung macrophage-epithelial cell interactions amplify particle-mediated cytokine release. AB - Interactions between alveolar macrophages (AMs) and epithelial cells may promote inflammatory responses to air pollution particles. Normal rat AMs, the alveolar type II epithelial cell line RLE-6TN (RLE), or cocultures of both cell types were incubated with various particles (0-50 microg/ml) for 24 h, followed by assay of released TNF-alpha and MIP-2. The particles used included titanium dioxide (TiO2), alpha-quartz (SiO2), residual oil fly ash (ROFA), or urban air particles (UAP). For all particles, a dose-dependent increase in TNF-alpha and MIP-2 release was observed in AM+RLE co-cultures but not in RLE or AM monoculture. AM+RLE co-culture also synergistically enhanced basal levels of tumor necrosis factor (TNF)-alpha and macrophage inflammatory protein (MIP)-2. In contrast, when AMs were co-cultured with fibroblasts, basal and particle-induced TNF-alpha and MIP-2 were similar to levels found in AM monoculture. Particle uptake by AMs was similar in mono- or AM+RLE co-culture. Increased basal and particle-induced cytokine release were not observed when the AMs were physically separated from the RLE. This contact-dependent cytokine potentiation could not be blocked with anti-CD18/anti-CD54, arginine-glycine-aspartate (RGD) peptide, or heparin. We conclude that in vitro inflammatory responses to particles are amplified by contact-dependent interactions between AMs and epithelial cells. AM-epithelial co culture may provide a useful model of in vivo particle effects. PMID- 11919088 TI - Cell cycle regulation of pulmonary phosphatidylcholine synthesis. AB - Pulmonary surfactant phosphatidylcholine (PC) formation increases as alveolar type II cells mature and arrest in G0/G1 state of the cell cycle at late fetal gestation. To determine whether this G0/G1 arrest is responsible for the increase in PC synthesis, we investigated the rates of PC synthesis and the activity, phosphorylation, intracellular distribution, synthesis, and degradation of a key enzyme of PC synthesis, cytidine triphosphate (CTP):phosphocholine cytidylyltransferase (CCTalpha). In synchronized mouse lung epithelial (MLE)-15 cells, PC production and CCTalpha activity peaked at G0/G1, declined during transition to G1/S, and remained low during S and G2/M. The changes in CCTalpha activity were not due to alterations in CCTalpha gene and protein expression. CCTalpha protein degradation also did not change during the cell cycle. Indirect immunofluorescence and immunogold electron microscopy revealed that CCTalpha localized to the cytoplasmic compartment and that its cytosolic localization did not change with the cell cycle. Although immunoblotting suggested no major redistribution of CCTalpha mass from cytosol to endoplasmic reticulum, activity measurements revealed that the ratio of particulate/soluble CCTalpha activity was cell cycle-dependent. The particulate/soluble ratio peaked at G0/G1 and declined with cell-cycle progression. Furthermore, the decrease in CCTalpha activity during exit from G0/G1 was associated with an increase in CCTalpha phosphorylation. These data suggest that the cell-cycle changes in PC synthesis are likely not due to alterations in CCTalpha expression and degradation but are primarily a consequence of changes in CCTalpha activity, phosphorylation, and membrane affinity. PMID- 11919089 TI - Community paediatrics and change. PMID- 11919091 TI - Fits, faints, or fatal fantasy? Fabricated seizures and child abuse. PMID- 11919092 TI - Air weapon injuries: a serious and persistent problem. PMID- 11919093 TI - How dangerous is food allergy in childhood? The incidence of severe and fatal allergic reactions across the UK and Ireland. AB - AIMS: To discover the incidence of fatal and severe allergic reactions to food in a large population of children. METHODS: A retrospective search for fatalities in children 0-15 years from 1990 to February 1998, primarily of death certification at offices of national statistics. A prospective survey of fatal and severe reactions from March 1998 to February 2000, primarily through the British Paediatric Surveillance Unit. MAIN OUTCOME MEASURES: were deaths and severe reactions. A case was deemed severe if one or more of the following criteria was met: cardiorespiratory arrest; need for inotropic support; fluid bolus >20 ml/kg; more than one dose of epinephrine; more than one dose of nebulised bronchodilator. A case was deemed near fatal if intubation was necessary. RESULTS: The UK under 16 population is 13 million. Over the past 10 years, eight children died (incidence of 0.006 deaths per 100 000 children 0-15 years per year). Milk caused four of the deaths. No child under 13 died from peanut allergy. Two children died despite receiving early epinephrine before admission to hospital; one child with a mild food allergic reaction died from epinephrine overdose. Over the past two years, there were six near fatal reactions (none caused by peanut) and 49 severe ones (10 caused by peanut), yielding incidences of 0.02 and 0.19 per 100 000 children 0-15 years per year respectively. Coexisting asthma is more strongly associated with a severe reaction than the severity of previous reactions. CONCLUSIONS: If 5% of the child population have food allergy, the risk that a food allergic child will die from a food allergic reaction is about 1 in 800 000 per year. The food allergic child with asthma may be at higher risk. Prescribing an epinephrine autoinjector requires a careful balance of advantages and disadvantages. PMID- 11919094 TI - Nut allergy in schoolchildren: a survey of schools in the Severn NHS Trust. AB - AIMS: To assess the extent of the problem of nut allergy in schoolchildren within the Severn NHS Trust. To determine how well informed schools are about the condition, their policies and attitudes, and the action that would be taken in the event of an acute reaction. METHODS: A questionnaire addressed to the head teacher was sent to 100 randomly selected mainstream schools in the Severn NHS Trust. RESULTS: Response rate was 83%. There were a total of 21 868 pupils in the schools. Forty five (54%) schools had at least one child currently known to be allergic. The total in all the schools was 87 (0.4%). Only 31 (36%) children had medication available in school. Of these, 18 (58%) had EpiPen alone. Twenty schools (44%) with an allergic child either had no staff trained to administer medication or did not respond to the question. Two (4%) schools with an allergic child had a support assistant for the pupils. Only 19 (43%) schools with a nut allergic child gave information to all teachers about nut allergy and only 21 (47%) gave information to dinner supervisors and other assistants. In only 23 (51%) schools with an allergic child were the cook and catering staff aware of all the children with a nut allergy. Ten (22%) schools with an allergic child served only "nut free dinners". Fourteen (31%) schools with a nut allergic child could not name a single sign of a mild acute allergic reaction (compared to 34 (89%) schools without an allergic child). Fifteen (33%) schools with an allergic child could not state a single sign of a severe acute allergic reaction (compared with 33 (87%) schools without a nut allergic child). CONCLUSION: Schools are not sufficiently well informed about nut allergy and management of acute allergic reactions. Policies and attitudes vary. We have revised the information given to schools regarding nut allergy and prepared a new information pack. PMID- 11919096 TI - More on erythromycin and pyloric stenosis. PMID- 11919095 TI - Six year effectiveness of a population based two tier infant hearing screening programme. AB - AIMS: To determine whether a two tier universal infant hearing screening programme (population based risk factor ascertainment and universal distraction testing) lowered median age of diagnosis of bilateral congenital hearing impairment (CHI) >40 dB HL in Victoria, Australia. METHODS: Comparison of whole population birth cohorts pre and post introduction of the Victorian Infant Hearing Screening Program (VIHSP). All babies surviving the neonatal period born in Victoria in 1989 (pre-VIHSP) and 1993 (post-VIHSP) were studied. (1) Pre-1992: distraction test at 7-9 months. (2) Post-1992: infants with risk factors for CHI referred for auditory brain stem evoked response (ABR) assessment; all others screened by modified distraction test at 7-9 months. RESULTS: Of the 1989 cohort (n = 63 454), 1.65/1000 were fitted with hearing aids for CHI by end 1995, compared with 2.09/1000 of the 1993 cohort (n = 64 116) by end 1999. Of these, 79 cases from the 1989 cohort (1.24/1000) and 72 cases from the 1993 cohort (1.12/1000) had CHI >40 dB HL. Median age at diagnosis of CHI >40 dB HL for the 1989 birth cohort was 20.3 months, and for the 1993 cohort was 14.2 months. Median age at diagnosis fell significantly for severe CHI but not for moderate or profound CHI. Significantly more babies with CHI >40 dB HL were diagnosed by 6 months of age in 1993 than in 1989 (21.7% v 6.3%). Compared to the six years pre VIHSP, numbers aided by six months were consistently higher in the six years post VIHSP (1.05 per 100 000 births versus 13.4 per 100 000 births per year). CONCLUSIONS: VIHSP resulted in very early diagnosis for more infants and lowered median age of diagnosis of severe CHI. However, overall results were disappointing. PMID- 11919097 TI - Family and parenting interventions for conduct disorder and delinquency: a meta analysis of randomised controlled trials. AB - AIMS: To determine whether family and parenting interventions benefit children and adolescents with conduct disorder and delinquency. METHODS: Meta-analysis of eight randomised controlled trials involving 749 children and adolescents (aged 10-17 years) with conduct disorder and/or delinquency. Criminality, academic performance, future employment, problem behaviour, family functioning, parental mental health, and peer relations were evaluated. RESULTS: Family and parenting interventions significantly reduced the time spent by juvenile delinquents in institutions (weighted mean difference 51.34 days). There was also a significant reduction in the risk of a juvenile delinquent being rearrested (relative risk 0.66) and in their rate of subsequent arrests at 1-3 years (standardised mean difference -0.56). CONCLUSIONS: The evidence suggests that family and parenting interventions for juvenile delinquents and their families have beneficial effects on reducing time spent in institutions and their criminal activity. In addition to the obvious benefit to the participant and their family, this may result in a cost saving for society. PMID- 11919099 TI - Resurgence of paediatric tuberculosis in London. PMID- 11919098 TI - Prevalence and secular trend of congenital anomalies in Glasgow, UK. AB - AIM: To describe the epidemiology of congenital anomalies in Glasgow with special reference to secular trends. METHODS: The prevalence of congenital anomalies was determined retrospectively in 233 777 births using the Glasgow Register of Congenital Anomalies for the period 1980-97. RESULTS: The total prevalence of congenital anomalies was 324 per 10 000 births, declining by just over a third from 382 per 10 000 births in 1980 to 238 per 10 000 births in 1997. The categories of defects with the highest prevalence were congenital heart disease (50 per 10 000 births), anomalies of limbs (49 per 10 000 births), and digestive system anomalies (47 per 10 000 births). Prevalence in most categories of anomaly declined, including those of the ear (-88%), congenital heart disease (-69%), anomalies of integument (-67%), nervous system anomalies (-61%), anomalies of limb (-54%), and urogenital (including renal) anomalies (-31%). By contrast, there was a significant upward trend for chromosomal anomalies (+50%). CONCLUSIONS: Despite the decline in the prevalence of many types of congenital anomaly, around 2.5% of all births in Glasgow were still associated with these disorders in 1997. In attempting to explain the prevalence and secular trend of congenital anomalies in Glasgow, underlying contributing factors require to be considered. These include changes in case ascertainment, antenatal screening, and diagnostic methods. PMID- 11919100 TI - Lipomatosis of the colon complicating Proteus syndrome. PMID- 11919101 TI - Why is management of patients with classical congenital adrenal hyperplasia more difficult at puberty? AB - Congenital adrenal hyperplasia (CAH) due to 21-hydroxylase deficiency is an autosomal recessive condition in which deletions or mutations of the cytochrome P450 21-hydroxylase gene cause glucocorticoid and often mineralocorticoid deficiency. Despite optimal substitution therapy, control of classical CAH is often inadequate at puberty, and the problems encountered relate to hypocortisolism and/or hyperandrogenism. A number of physiological alterations in the endocrine milieu at puberty, which include alterations in the growth hormone/insulin-like growth factor axis, insulin sensitivity, as well as the activity of enzymes participating in cortisol metabolism and adrenal steroidogenesis, may account for the documented hypocortisolism and elevated androgen production, and may explain the difficulty in maintaining adequate adrenocortical suppression in pubertal patients with classical 21-hydroxylase deficiency. PMID- 11919102 TI - Cough, airway inflammation, and mild asthma exacerbation. AB - BACKGROUND: Prospective data on the temporal relation between cough, asthma symptoms, and airway inflammation in childhood asthma is unavailable. AIMS AND METHODS: Using several clinical (diary, quality of life), lung function (FEV(1), FEV(1) variability, airway hyperresponsiveness), cough (diary, cough receptor sensitivity (CRS)), and inflammatory markers (sputum interleukin 8, eosinophilic cationic protein (ECP), myeloperoxidase; and serum ECP) of asthma severity, we prospectively described the course of these markers in children with asthma during a non-acute, acute, and resolution phase. A total of 21 children with asthma underwent these baseline tests; 11 were retested during days 1, 3, 7, and 28 of an exacerbation. RESULTS: Asthma exacerbations were characterised by increased asthma and cough symptoms and eosinophilic inflammation. Sputum ECP showed the largest increase and peaked later than clinical scores. Asthma scores consistently related to cough score only early in the exacerbation. Neither CRS nor cough scores related to any inflammatory marker. CONCLUSION: In mild asthma exacerbations, eosinophilic inflammation is dominant. In asthmatic children who cough as a dominant symptom, cough heralds the onset of an exacerbation and increased eosinophilic inflammation, but cough scores and CRS do not reflect eosinophilic airway inflammation. PMID- 11919103 TI - Screening for coeliac disease has benefits in Williams syndrome. PMID- 11919105 TI - Identifying risk of blindness. PMID- 11919104 TI - Short term effects of adrenaline in bronchiolitis: a randomised controlled trial. AB - BACKGROUND: Airway narrowing in acute bronchiolitis does not respond to inhaled bronchodilators but does to adrenaline when compared to bronchodilators. Influences of supportive care were not considered in previous treatment studies. METHODS: Short term effects of nebulised adrenaline and saline placebo were compared in infants with moderately severe acute bronchiolitis. Thirty eight infants were recruited, 19 in each treatment group. After stabilisation, infants received a single 3 ml dose of either levo-adrenaline (3 mg) or 0.9% saline placebo via Pari-BABY nebuliser driven with 6 l/min oxygen for three minutes. Changes in respiratory rate (RR), heart rate (HR), oxygen saturation (SpO(2)), Respiratory Distress Assessment Instrument (RDAI), and activity levels were assessed at 20 minutes intervals at times -20, 0, 20, 40, and 60 minutes around treatment. Respiratory virology and chest x ray were performed. RESULTS: Supportive therapy prior to study treatment resulted in significant reductions in RR (by 4.3 breaths/min) and HR (by 4.6 beats/min); there were no changes in SpO(2) or RDAI. There were no further changes in any parameter in either treatment group at any assessment time after treatment. CONCLUSION: No improvement was shown with inhaled adrenaline in acute bronchiolitis, when compared with supportive care or placebo. Improvements noted pretreatment question whether prior noted improvements were through supportive care or pharmacological interventions. PMID- 11919106 TI - Differential response of wheezes and ruttles to anticholinergics. AB - Computerised breath sounds analysis was used to assess the response of infants with wheeze and ruttles to the drug ipratropium bromide. A mean decrease in breath sounds intensity occurred in infants with ruttles after five minutes (20.5 dB), but not until 20 minutes in those with wheeze (8.1 dB). This differential response may be related to different underlying pathophysiology. PMID- 11919107 TI - Procalcitonin as a diagnostic marker of meningococcal disease in children presenting with fever and a rash. AB - BACKGROUND: Procalcitonin (PCT), a precursor of calcitonin, is a recognised marker of bacterial sepsis, and high concentrations correlate with the severity of sepsis. PCT has been proposed as an earlier and better diagnostic marker than C reactive protein (CRP) and white cell count (WCC). This comparison has never been reported in the differentiation of meningococcal disease (MCD) in children presenting with a fever and rash. AIM: To determine if PCT might be a useful marker of MCD in children presenting with fever and rash. METHODS: PCT, CRP, and WCC were measured on admission in 108 children. Patients were classified into two groups: group I, children with a microbiologically confirmed clinical diagnosis of MCD (n = 64); group II, children with a self limiting illness (n = 44). Median ages were 3.57 (0.07-15.9) versus 1.75 (0.19-14.22) years respectively. Severity of disease in patients with MCD was assessed using the Glasgow Meningococcal Septicaemia Prognostic Score (GMSPS). RESULTS: PCT and CRP values were significantly higher in group I than in group II (median 38.85 v 0.27 ng/ml and 68.35 v 9.25 mg/l; p < 0.0005), but there was no difference in WCC between groups. Sensitivity, specificity, and positive and negative predictive values were higher for PCT than CRP and WCC. In group I, procalcitonin was significantly higher in those with severe disease (GMSPS >/=8). CONCLUSIONS: PCT is a more sensitive and specific predictor of MCD than CRP and WCC in children presenting with fever and a rash. PMID- 11919109 TI - Ultrafine particles: insidious invaders. PMID- 11919108 TI - Kawasaki disease: an evidence based approach to diagnosis, treatment, and proposals for future research. AB - This article proposes a clinical guideline for the diagnosis and treatment of Kawasaki disease in the UK based on the best available evidence to date, and highlights areas of practice where evidence is anecdotal or based on retrospective data. Future research as proposed by the London Kawasaki Disease Research Group is outlined, and clinicians are invited to prospectively enroll their suspected cases into this collaborative research project. PMID- 11919110 TI - Prevalence and distribution of petechiae in well babies. AB - AIMS: To determine the prevalence of petechial spots in well babies. METHODS: A total of 116 babies under the age of 12 months were fully examined at child health surveillance clinics. The number and site of petechiae were recorded together with details of possible causes. RESULTS: A total of 27.6% of babies had one or more petechiae, 8.6% had two or more petechiae, and 2.6% had more than two. None of these babies subsequently developed sepsis. CONCLUSIONS: Many well infants examined in the community are likely to have petechial spots. In this setting one or two petechiae are common and their presence should not be taken as pathological without other clinical signs. Recognition of this fact may also be helpful when examining otherwise well infants with petechiae in a secondary care setting. PMID- 11919111 TI - Extreme variability of expression of a Sonic Hedgehog mutation: attention difficulties and holoprosencephaly. AB - Holoprosencephaly (HPE) is a clinically variable and genetically heterogeneous central nervous system (CNS) malformation. Alobar HPE, which is its most severe form, is associated with a poor prognosis. At the milder end of the HPE spectrum microcephaly, hypotelorism, and single central maxillary incisor may be recognised. Currently, four genes have been identified for this condition. These include Sonic Hedgehog (SHH) on chromosome 7q36, which is thought to be responsible for a significant proportion of autosomal dominant HPE. We report an index case with alobar holoprosencephaly caused by an SHH mutation and six members of his family over two generations with this mutation, with a broad range of clinical presentation, including attention deficit hyperactivity disorder (ADHD). The combination of microcephaly, hypotelorism, subtle midline facial anomalies, and ADHD within a sibship should alert the physician to the possible diagnosis of HPE. PMID- 11919112 TI - Hermansky-Pudlak syndrome: infrequent bleeding and first report of Turkish and Pakistani kindreds. AB - Hermansky-Pudlak syndrome (HPS) is a rare disorder characterised by oculocutaneous albinism, a bleeding tendency, and lipofuscinosis. This retrospective study reviews the clinical history and haematological features of 23 cases of HPS. Information was gathered from patient notes and by direct interview. Thirteen of the 23 children were of Turkish origin, 12 being members of four kindreds from the Turkish/Kurdish border. Four children originated from Pakistan. Haemorrhage was uncommon; two experienced significant bleeding (intracranial and retinal haemorrhage in one and menorrhagia in another), and twelve minor symptoms. Results of laboratory evaluation of platelet function were not predictive of bleeding; in particular the PFA-100 analyser was not sensitive to the HPS defect. The most sensitive test of platelet fuction was quantitation of platelet nucleotides. The occurrence of Turkish and Pakistani kindreds with HPS is novel and follow up for long term complications described in Puerto Rican patients as well as genetic analysis is ongoing. PMID- 11919113 TI - Right atrial isomerism. PMID- 11919114 TI - Pulmonary oedema caused by "liquid ecstasy" ingestion. AB - In this case report we describe the first recorded case of alveolar gas exchange impairment caused by "liquid ecstasy" ingestion, and perhaps inhalation, by a 4 year old child. The pulmonary gas diffusion disturbance was sufficiently prolonged to raise the suspicion of a direct toxic effect on the alveolar capillary membrane. PMID- 11919115 TI - Guillain-Barre syndrome associated with central nervous system lesions. AB - We report the clinical course, and neurophysiological and neuroimaging findings of a patient with Guillain-Barre syndrome associated with central nervous system lesions. During a course of intravenous immunoglobulin therapy, she had headache with meningism. Cerebral magnetic resonance imaging showed lesions in both frontal and right occipital lobes. Cerebrospinal fluid showed a raised protein concentration accompanied by mild pleocytosis. Her symptoms resolved within two months. Subsequent magnetic resonance imaging revealed cavity formation in the deep white matter and atrophic changes in the right occipital lobes. PMID- 11919116 TI - Polycythaemia and hypertension caused by renal artery stenosis. AB - A girl with failure to thrive and a haemoglobin of 140 g/l at 1.3 years died from a brain haemorrhage 2.5 years later. Renal artery stenosis had caused severe, chronic hypertension and increased erythropoietin secretion (haemoglobin 182 g/l). Blood pressure should be measured in all unwell children, including those failing to thrive. PMID- 11919117 TI - Rapid skin anaesthesia using high velocity lignocaine particles: a prospective placebo controlled trial. AB - BACKGROUND: Local anaesthetic creams (EMLA and Ametop) are used widely to provide pain free intravenous cannulation. However, they take a minimum of 45 minutes to become effective. AIMS: To evaluate a prototype device, dermal Powderject lidocaine (DPL), that delivers high velocity lignocaine particles into the skin. METHODS: A total of 132 children (aged 4-12 years) were randomised to receive either a sham delivery or a delivery of DPL on the skin at the antecubital fossa, or back of hand. Pain of intravenous cannulation was assessed four minutes later using self reporting behaviours and blinded observation with standard pain assessment tools. The trial was designed to measure both efficacy of skin anaesthesia and potential skin damage with increasing driving pressure of the device (30 or 40 bar), and different lignocaine particle sizes (<38 micrometer or 38-53 micrometer) in a block randomised fashion. RESULTS: A total of 128 patients were evaluable. There was a trend towards improved anaesthesia at higher device pressures at the antecubital fossa with both self reporting and blinded observation. Acceptable analgesia was achieved in 90% of patients for high pressure at both particle sizes compared to 60% and 40% for the sham device using self reporting measures. The observed differences using the blinded observer were similar: 90% v 20% (40 bar and small particles v sham), and 80% v 40% (40 bar and large particles v sham). At the back of hand the differences between active and sham devices were not significant. The device was well tolerated and not associated with pain on deployment. One patient had mild petechiae and oedema after deployment (Draize score of 3). CONCLUSIONS: This prototype device appears to provide significant skin anaesthesia at the antecubital fossa, but not at the back of hand. The device is not painful to use and causes only minor short term skin changes. PMID- 11919119 TI - Effect of a chicken-based diet on renal function and lipid profile in patients with type 2 diabetes: a randomized crossover trial. AB - OBJECTIVE: To assess the effect of replacing red meat with chicken in the usual diet and the effect of a low-protein diet on glomerular filtration rate (GFR), urinary albumin excretion rate (UAER), and lipid levels in patients with type 2 diabetes. RESEARCH DESIGN AND METHODS: A randomized, crossover, controlled trial was conducted with 28 patients with type 2 diabetes (seven women; mean age 58.1 years): 15 patients were normoalbuminuric (UAER <20 microg/min), and 13 patients were microalbuminuric (UAER 20-200 microg/min). A chicken-based diet (red meat replaced with chicken) and a low-protein diet were compared with the patients' usual diet. Patients followed each diet for 4 weeks with a 4-week washout period between. GFR ((51)Cr-EDTA single-injection technique), 24-h UAER (immunoturbidimetry), apolipoprotein B, total cholesterol, LDL cholesterol, HDL cholesterol, and triglycerides were measured after each diet. RESULTS: Normoalbuminuric and microalbuminuric patients with diabetes were analyzed separately. In normoalbuminuric patients, GFR after the chicken (101.3 +/- 22.9 ml x min(-1) x 1.73 m(-2)) and low-protein diets (93.8 +/- 20.5 ml x min(-1) x m( 2)) was lower than after the usual diet (113.4 +/- 31.4 ml x min(-1) x 1.73 m( 2); P < 0.05). In microalbuminuric patients, apolipoprotein B levels were lower after the chicken (113.5 +/- 36.0 mg/dl) and low-protein diets (103.5 +/- 40.1 mg/dl) than after the usual diet (134.3 +/- 30.7 mg/dl; P < 0.05). Only the chicken diet reduced UAER (median 34.3 microg/min) compared with the low-protein (median 52.3 microg/min) and usual (median 63.8 microg/min) diets (P < 0.05). Glycemic control and blood pressure did not change after the diets. CONCLUSIONS: A normoproteic diet with chicken as the only source of meat may represent an alternative strategy for treatment of patients with type 2 diabetes and microalbuminuria. PMID- 11919120 TI - Effects of energy-restricted diets containing increased protein on weight loss, resting energy expenditure, and the thermic effect of feeding in type 2 diabetes. AB - OBJECTIVE: To determine the effect of a high-protein (HP) diet compared with a low-protein (LP) diet on weight loss, resting energy expenditure (REE), and the thermic effect of food (TEF) in subjects with type 2 diabetes during moderate energy restriction. RESEARCH DESIGN AND METHODS: In this study, 26 obese subjects with type 2 diabetes consumed a HP (28% protein, 42% carbohydrate) or LP diet (16% protein, 55% carbohydrate) during 8 weeks of energy restriction (1,600 kcal/day) and 4 weeks of energy balance. Body weight and composition and REE were measured, and the TEF in response to a HP or LP meal was determined for 2 h, at weeks 0 and 12. RESULTS: The mean weight loss was 4.6 +/- 0.4 kg (P < 0.001), of which 4.5 +/- 0.4 kg was fat (P < 0.001), with no effect of diet (P = 0.6). At both weeks 0 and 12, TEF was greater after the HP than after the LP meal (0.064 vs. 0.050 kcal x kcal(-1) energy consumed x 2 h(-1), respectively; overall diet effect, P = 0.003). REE and TEF were reduced similarly with each of the diets (time effects, P = 0.02 and P < 0.001, respectively). CONCLUSIONS: In patients with type 2 diabetes, a low-fat diet with an increased protein-to-carbohydrate ratio does not significantly increase weight loss or blunt the fall in REE. PMID- 11919121 TI - Effects of dietary treatment alone or diet with voglibose or glyburide on abdominal adipose tissue and metabolic abnormalities in patients with newly diagnosed type 2 diabetes. AB - OBJECTIVE; To examine the effects of diet and diet with voglibose or glyburide on abdominal adiposity and metabolic abnormalities in patients with type 2 diabetes. RESEARCH DESIGN AND METHODS: A total of 36 Japanese patients with newly diagnosed type 2 diabetes (50.8 +/- 8.6 years of age, BMI 24.5 +/- 3.5 kg/m(2)) and 273 normal control subjects were studied. The patients were treated for 3 months with diet alone (30 kcal/kg per day) (n = 15), diet with voglibose (n = 12), or diet with glyburide (n = 9). They underwent 75-g oral glucose tolerance testing, assessment of insulin sensitivity (SI), and acute insulin response (AIR) with intravenous glucose tolerance testing based on the minimal model, and measurement of abdominal visceral adipose tissue area (VAT) and subcutaneous adipose tissue area (SAT) by computed tomography before and after treatment. RESULTS: The diabetic patients had comparable SAT but larger VAT than the control subjects. With a mean weight loss of 2-3 kg, VAT and SAT were decreased similarly in all treatment groups. The VAT-to-SAT ratio was decreased only in the voglibose group. Glycemic control and serum lipid profiles were improved in all groups. Changes in glycemic control after diet were closely correlated with changes in VAT but not with changes in SAT. SI and AIR were unchanged in the diet group but were improved in the voglibose and glyburide groups. CONCLUSIONS: In Japanese patients with newly diagnosed type 2 diabetes who were relatively lean but had excess VAT, diet with or without voglibose or glyburide effectively reduced VAT. Decrease in VAT was closely associated with improvement of glycemic control with diet. Additional use of voglibose or low-dose glyburide had no detrimental effects on abdominal adiposity and had beneficial effects on SI and AIR. PMID- 11919122 TI - A low-sodium diet potentiates the effects of losartan in type 2 diabetes. AB - OBJECTIVE: Diabetic subjects have a high prevalence of hypertension, increased total body exchangeable sodium levels, and an impaired ability to excrete a sodium load. This study assessed the effect of dietary sodium restriction on the efficacy of losartan in hypertensive subjects with type 2 diabetes and albumin excretion rates of 10-200 microg/min. RESEARCH DESIGN AND METHODS: In this study, 20 subjects were randomized to losartan 50 mg/day (n = 10) or placebo (n = 10). Drug therapy was given in two 4-week phases separated by a washout period. In the last 2 weeks of each phase, patients were assigned to low- or regular-sodium diets, in random order. In each phase, 24-h ambulatory blood pressure, urinary albumin-to-creatinine ratio (ACR), and renal hemodynamics were measured. RESULTS: Achieved urinary sodium on a low-sodium diet was 85 +/- 14 and 80 +/- 22 mmol/day in the losartan and placebo groups, respectively. In the losartan group, the additional blood pressure-lowering effects of a low-sodium diet compared with a regular-sodium diet for 24-h systolic, diastolic, and mean arterial blood pressures were 9.7 mmHg (95% confidence interval [CI], 2.2-17.2; P = 0.002), 5.5 mmHg (2.6-8.4; P = 0.002), and 7.3 mmHg (3.3- 11.3; P = 0.003), respectively. In the losartan group, the ACR decreased significantly on a low-sodium diet versus on a regular-sodium diet (-29% [CI -50.0 to -8.5%] vs. + 14% [-19.4 to 47.9%], respectively; P = 0.02). There was a strong correlation between fall in blood pressure and percent reduction in the ACR (r = 0.7, P = 0.02). In the placebo group, there were no significant changes in blood pressure or ACR between regular and low-sodium diets. There were no significant changes in renal hemodynamics in either group. CONCLUSIONS: These data demonstrated that a low-sodium diet potentiates the antihypertensive and antiproteinuric effects of losartan in type 2 diabetes. The blood pressure reduction resulting from the addition of a low sodium diet to losartan was of similar magnitude to that predicted from the addition of a second antihypertensive agent. PMID- 11919123 TI - Sexual dysfunction in women with type 1 diabetes: a controlled study. AB - OBJECTIVE: This study aimed to 1) examine the prevalence of sexual problems in women with type 1 diabetes, 2) compare this prevalence rate with that of an age matched control group, 3) study the influence of diabetes-related somatic factors on female sexuality, and 4) study the influence of psychological variables on the sexual functioning of both groups. RESEARCH DESIGN AND METHODS: A total of 120 women with diabetes visiting the outpatient diabetes clinic completed questionnaires evaluating psychological adjustment to diabetes, marital satisfaction, depression, and sexual functioning. Medical records were used to obtain data on HbA(1c), use of medication, BMI, and early-onset microvascular complications. An age-matched control group of 180 healthy women attending an outpatient gynecological clinic for preventive routine gynecological assessment also completed the non-diabetes-related questionnaires. RESULTS: More women with diabetes than control subjects reported sexual dysfunction (27 vs. 15%; P = 0.04), but a significant difference was found only for decreased lubrication. No association was found between sexual dysfunction and age, BMI, duration of diabetes, HbA(1c), use of medication, menopausal status, or complications. Women with more complications, however, reported significantly more sexual dysfunctions, and the presence of complications altered treatment satisfaction. Both diabetic and control women with sexual dysfunction mentioned lower overall quality of the marital relation and more depressive symptoms than their respective counterparts without sexual problems. Depression was a significant predictor for sexual dysfunction in both women with diabetes and control subjects. CONCLUSIONS: Sexual problems are frequent in women with diabetes. They affect the overall quality of life and deserve more attention in clinical practice and research. PMID- 11919124 TI - Comorbidities and impairments explaining the association between diabetes and lower extremity disability: The Women's Health and Aging Study. AB - OBJECTIVE: To elucidate the role of diabetes-related impairments and comorbidities in the association between diabetes and physical disability, this study examined the association between diabetes and lower extremity function in a sample of disabled older women. RESEARCH DESIGN AND METHODS: Cross-sectional analysis of 1,002 women (aged >or=65 years) enrolled in the Women's Health and Aging Study (one-third most disabled of the total community-dwelling population). Diabetes and other medical conditions were ascertained by standard criteria that used multiple sources of information. Functional status was assessed using self reported and objective performance measures. RESULTS: Women with diabetes were significantly more likely to have cardiovascular diseases, peripheral nerve dysfunction, visual impairment, obesity, and depression. After adjustment for age, women with diabetes had a greater prevalence of mobility disability (odds ratio [OR] 1.85, 95% CI 1.12-3.06), activities of daily living disability (1.61, 1.06-2.43), and severe walking limitation (2.34, 1.56-3.50), and their summary mobility performance score (0-12 scale based on balance, gait speed, chair stands) was 1.4 points lower than in nondiabetic women (P < 0.001). Peripheral artery disease, peripheral nerve dysfunction, and depression were the main individual contributing factors; however, none of these conditions alone fully explained the association between diabetes and disability. Conversely, only after adjusting for all potential mediators was the relationship between diabetes and disability reduced to a large degree. CONCLUSIONS: Even among physically impaired older women, diabetes is associated with a major burden of disability. A wide range of impairments and comorbidities explains the diabetes-disability relationship, suggesting that the mechanism for such an association is multifactorial. PMID- 11919125 TI - Does diabetes disease management save money and improve outcomes? A report of simultaneous short-term savings and quality improvement associated with a health maintenance organization-sponsored disease management program among patients fulfilling health employer data and information set criteria. AB - OBJECTIVE: Little is known about the impact of disease management programs on medical costs for patients with diabetes. This study compared health care costs for patients who fulfilled health employer data and information set (HEDIS) criteria for diabetes and were in a health maintenance organization (HMO) sponsored disease management program with costs for those not in disease management. RESEARCH DESIGN AND METHODS: We retrospectively examined paid health care claims and other measures of health care use over 2 years among 6,799 continuously enrolled Geisinger Health Plan patients who fulfilled HEDIS criteria for diabetes. Two groups were compared: those who were enrolled in an opt-in disease management program and those who were not enrolled. We also compared HEDIS data on HbA(1c) testing, percent not in control, lipid testing, diabetic eye screening, and kidney disease screening. All HEDIS measures were based on a hybrid method of claims and chart audits, except for percent not in control, which was based on chart audits only. RESULTS: Of 6,799 patients fulfilling HEDIS criteria for the diagnosis of diabetes, 3,118 (45.9%) patients were enrolled in a disease management program (program), and 3,681 (54.1%) were not enrolled (nonprogram). Both groups had similar male-to-female ratios, and the program patients were 1.4 years younger than the nonprogram patients. Per member per month paid claims averaged 394.62 dollars for program patients compared with 502.48 dollars for nonprogram patients (P < 0.05). This difference was accompanied by lower inpatient health care use in program patients (mean of 0.12 admissions per patient per year and 0.56 inpatient days per patient per year) than in nonprogram patients (0.16 and 0.98, P < 0.05 for both measures). Program patients experienced fewer emergency room visits (0.49 per member per year) than nonprogram patients (0.56) but had a higher number of primary care visits (8.36 vs. 7.78, P < 0.05 for both measures). Except for emergency room visits, these differences remained statistically significant after controlling for age, sex, HMO enrollment duration, presence of a pharmacy benefit, and insurance type. Program patients also achieved higher HEDIS scores for HbA(1c) testing as well as for lipid, eye, and kidney screenings (96.6, 91.1, 79.1, and 68.5% among program patients versus 83.8, 77.6, 64.9, and 39.3% among nonprogram patients, P < 0.05 for all measures). Among 1,074 patients with HbA(1c) levels measured in a HEDIS chart audit, 35 of 526 (6.7%) program patients had a level >9.5%, as compared with 79 of 548 (14.4%) nonprogram patients. CONCLUSIONS: In this HMO, an opt-in disease management program appeared to be associated with a significant reduction in health care costs and other measures of health care use. There was also a simultaneous improvement in HEDIS measures of quality care. These data suggest that disease management may result in savings for sponsored managed care organizations and that improvements in HEDIS measures are not necessarily associated with increased medical costs. PMID- 11919126 TI - Association of parity with risk of type 2 diabetes and related metabolic disorders. AB - OBJECTIVE: The relationship between parity and risk of diabetes is controversial, and little information is available regarding associations between parity and measures of insulin resistance and beta-cell function. The objective of this study was to investigate the association between parity and risk of glucose intolerance and related metabolic disorders using data from a population-based study in a Native Canadian community. RESEARCH DESIGN AND METHODS: Female participants (n = 383, aged 12-79 years) provided fasting blood samples for the determination of glucose, insulin, C-peptide, and proinsulin concentrations. A 75 g oral glucose tolerance test was administered, and diabetes and impaired glucose tolerance were diagnosed according to World Health Organization criteria. Waist circumference and percent body fat were determined. Information regarding occurrence of live births and previously diagnosed diabetes was obtained from interviewer-administered questionnaires. RESULTS: Parity was associated with a significantly reduced risk of diabetes (nulliparous vs. >or=1 birth, odds ratio 0.43, 95% CI 0.19- 0.94, P < 0.05) after adjustment for age and waist circumference. In addition, nondiabetic nulliparous women had significantly elevated concentrations of fasting insulin and proinsulin relative to nondiabetic parous women (all P < 0.05) in analyses adjusted for age and waist circumference. CONCLUSIONS: Our results are consistent with those from other populations experiencing high rates of diabetes and suggest the presence of a diabetes-prone phenotype within the nulliparous subcohort of this population, which may contribute to infertility. PMID- 11919127 TI - Impact of diabetes on coronary stenosis and coronary artery calcification detected by electron-beam computed tomography in symptomatic patients. AB - OBJECTIVE: Ischemic heart disease is a pivotal complication for diabetic patients. Electron-beam computed tomography (EBCT) represents the only noninvasive method that allows for accurate quantification of coronary artery calcification that reflects underlying atherosclerotic disease. Although coronary calcium score (CCS) cut points that predict the presence of angiographic stenosis have been established in nondiabetic individuals, it is not known whether coronary calcifications in diabetic patients are associated with the presence of significant coronary stenoses. In this study, we evaluated the relationship between coronary calcifications and angiographic stenosis in symptomatic patients with or without type 2 diabetes. RESEARCH DESIGN AND METHODS: In this study, 282 patients (204 men and 78 women) with chest pain, including 101 diabetic patients and 181 nondiabetic patients (mean age 63 +/- 9.6 years), underwent coronary angiography and EBCT with determination of CCS using Agatston's method. Luminal stenosis >or= 50% was defined as significant coronary stenosis. RESULTS: Angiography identified 205 patients with significant stenoses (89 of 101 diabetic patients, 114 of 181 nondiabetic patients). The sensitivity and specificity of EBCT to detect significant coronary stenosis were not significantly different between diabetic and nondiabetic patients. In diabetic patients, a CCS >or=90 was associated with 75% sensitivity and 75% specificity, whereas a CCS >or=200 was associated with 64% sensitivity and 83% specificity. CONCLUSIONS: We demonstrated that calcification of the coronary arteries in symptomatic diabetic patients is well associated with severity of coronary stenosis, as in nondiabetic patients. PMID- 11919128 TI - Prevalence of Q-T interval dispersion in type 1 diabetes and its relation with cardiac ischemia : the EURODIAB IDDM Complications Study Group. AB - OBJECTIVE: The interlead variation in duration of the Q-T interval on the surface electrocardiogram (Q-T interval dispersion [QTd]) has been shown to predict mortality in type 2 diabetic patients. We evaluated the prevalence of QTd prolongation in the EURODIAB population and its relation to corrected Q-T interval (QTc), sex, age, duration of diabetes, blood glucose control, and complications. RESEARCH DESIGN AND METHODS; A total of 3,042 type 1 diabetic patients were studied. QTc was calculated according to the Bazett's formula; QTc > 0.44 s was considered abnormally prolonged. QTd was calculated using the difference between the maximum and the minimum QTc in any thoracic lead. QTd >0.080 s was considered abnormally prolonged. RESULTS: The prevalence of an increased QTd was 7%. A significant relation was observed between QTd prolongation and diastolic blood pressure (P < 0.05). A higher prevalence of QTd prolongation was observed in patients with ischemic heart disease (P = 0.004), whereas no relationship was observed with retinopathy, albumin excretion rate, or measures of somatic and autonomic neuropathy. QTc and QTd were significantly related (P = 0.001); however, a proportion of patients with normal QTd showed a prolonged QTc (>0.44 s). CONCLUSIONS: In patients with type 1 diabetes, QTd is associated with ischemic heart disease and diastolic blood pressure but not neuropathy. Although QTd is statistically related to duration of QTc, increased QTd and increased QTc identify different patients, and their predictive value deserves prospective evaluation. PMID- 11919129 TI - A prospective, randomized comparison of the metabolic effects of pioglitazone or rosiglitazone in patients with type 2 diabetes who were previously treated with troglitazone. AB - OBJECTIVE: To characterize potential differences in glycemic control, plasma lipid level, and weight in a cohort of patients previously treated with troglitazone (TROG) who were switched to either pioglitazone or rosiglitazone. RESEARCH DESIGN AND METHODS: After a 2-week washout from TROG, 186 patients were randomly assigned to receive either pioglitazone (PIO) or rosiglitazone (ROSI). Weight, HbA(1c), and fasting lipid profile were documented before discontinuing TROG and at 4 months after starting either pioglitazone or rosiglitazone. Secondarily, the effect of concurrent medications on study outcomes was assessed. RESULTS: A total of 127 patients completed follow-up: 67 individuals in the PIO group (32 women, 35 men) and 60 individuals in the ROSI group (33 women, 27 men). There were no significant differences in gender mix, age, weight, fasting lipid profile, or HbA(1c) between the ROSI and PIO groups. After 4 months of randomized treatment, no change in HbA(1c) from baseline between or within groups was noted. Both groups experienced an equal and significant increase in weight from baseline of approximately 2.0 kg. Thiazolidinedione and HMG-CoA reductase inhibitor therapy had significant and independent effects on lipid profile (P < 0.005). Significant improvements in lipid profile were noted in the PIO group (P < 0.01), whereas none were detected with conversion to ROSI. Specifically, the PIO group experienced an average decrease in total cholesterol of approximately 20 mg/dl. CONCLUSIONS: Differing effects on lipid profile were apparent after random conversion from TROG to either PIO or ROSI, despite similar weight increase and glycemic control. The clinical significance of these differences remains to be determined, and further comparative research is warranted. PMID- 11919130 TI - Additional lunchtime basal insulin during insulin lispro intensive therapy in a randomized, multicenter, crossover study in adults : a real-life design. AB - OBJECTIVE: This study was performed to evaluate whether an additional dose of NPH insulin at lunchtime might overcome the deleterious effects of waning basal insulinemia on pre-dinner and evening glucose values during insulin lispro intensive therapy with once daily basal insulin at night. RESEARCH DESIGN AND METHODS: The study was a 10-month multicenter, randomized, crossover trial. After a 2-month run-in period, subjects injected NPH insulin once (1 x NPH) or twice (2 x NPH) daily for 4 months in a randomized order. Adult patients were included if they had HbA(1c) levels <8.5%. Efficacy measures were HbA(1c) levels, 8-point glucose profiles, and the frequency of hypoglycemia. The statistical analysis included a within-patient comparison for crossover trials. RESULTS: In all, 104 patients completed the trial. The mean HbA(1c) level before randomization was 7.1 +/- 0.85%. The HbA(1c) levels did not change significantly within patients (t test, mean difference = 0.06%; 95% confidence interval [CI] -0.073 to 0.20). The pre-dinner blood glucose values were significantly lower during the 2 x NPH daily protocol, with a mean difference of 0.76 mmol/l (t test, P = 0.004; CI 0.25 to 1.3). In the evening, the frequency of hypoglycemia increased significantly during the 2 x NPH daily protocol with a median difference of 0.56 mild episodes/30 days (P = 0.001) and 6.9 severe episodes/patient year (P = 0.007), respectively. CONCLUSIONS: Equal HbA(1c) levels and increasing frequencies of hypoglycemia in the evening overshadow the slight improvement of the evening glucose profiles during a regimen with 2 x NPH daily insulin. Therefore, generalized use of a second injection of NPH insulin at lunchtime cannot be recommended to all adult patients with type 1 diabetes using intensive insulin lispro therapy. PMID- 11919131 TI - Control of cardiovascular risk factors in patients with diabetes and hypertension at urban academic medical centers. AB - OBJECTIVE: There are national mandates to reduce blood pressure (BP) to <130/85 mmHg, LDL cholesterol to <100 mg/dl, and HbA(1c) to <7% and to institute aspirin therapy in patients with diabetes. The objective of this study was to determine the proportion of patients in urban institutions with diabetes and hypertension who meet these treatment goals. RESEARCH DESIGN AND METHODS: Using American Diabetes Association (ADA) guidelines, we evaluated the control of cardiovascular disease (CVD) risk factors in 1,372 patients receiving medical care at two major urban medical centers in Brooklyn and Detroit. Information was extracted from charts of outpatient clinics. RESULTS: Of 1,372 active clinic patients with diabetes and hypertension, 1,247 (90.9%) had type 2 diabetes, and 26.7% met the target blood pressure of 130/85 mmHg. A total of 35.5% met the goal LDL cholesterol level of <100 mg/dl, 26.7% had an HbA(1c) <7%, and 45.6% were on antiplatelet therapy. Only 3.2% of patients met the combined ADA goal for BP, LDL cholesterol, and HbA(1c). CONCLUSIONS: Optimal control of CVD risk factors in adults with diabetes was achieved only in a minority of patients. Results reflect the inherent difficulties in achieving these complex guidelines in our present health care systems. PMID- 11919132 TI - A randomized study and open-label extension evaluating the long-term efficacy of pramlintide as an adjunct to insulin therapy in type 1 diabetes. AB - OBJECTIVE: To assess the effect of mealtime amylin replacement with pramlintide on long-term glycemic and weight control in patients with type 1 diabetes. RESEARCH DESIGN AND METHODS: In a 52-week, double-blind, placebo-controlled, multicenter study, 480 patients with type 1 diabetes were randomized to receive preprandial injections of placebo or 30 microg pramlintide q.i.d., in addition to existing insulin regimens. At week 20, pramlintide-treated patients were re randomized to 30 or 60 microg pramlintide q.i.d. if decreases from baseline in HbA(1c) were <1% at week 13. Of the 342 patients who completed the 52-week study, 236 individuals ( approximately 70%) elected to participate in a 1-year open label extension in which all patients received 30 or 60 microg pramlintide q.i.d. RESULTS: Treatment with pramlintide led to a mean reduction in HbA(1c) of 0.67% from baseline to week 13 that was significantly (P < 0.0001) greater than the placebo reduction (0.16%), and a significant placebo-corrected treatment difference was sustained through week 52 (P = 0.0071). The greater HbA(1c) reduction was associated with an average weight loss, rather than weight gain, and was not accompanied by an increased overall event rate of severe hypoglycemia. In the open-label extension, mean HbA(1c) levels decreased rapidly in patients receiving pramlintide for the first time and remained at reduced levels in patients who continued pramlintide treatment. The most common adverse events reported by the pramlintide group were mild nausea and anorexia, which both occurred during the initial weeks of treatment and dissipated over time. CONCLUSIONS: Mealtime pramlintide treatment as an adjunct to insulin improved long-term glycemic control without inducing weight gain or increasing the overall risk of severe hypoglycemia in patients with type 1 diabetes. PMID- 11919133 TI - Interpersonal predictors of HbA(1c) in patients with type 1 diabetes. AB - OBJECTIVE: Research suggests that increased collaboration and satisfaction in the patient-provider relationship is associated with better outcomes in patients with diabetes. In adults, an interpersonal style characterized by low trust of others and excessive self-reliance is known as "dismissing attachment style." We hypothesized that diabetic patients with dismissing attachment style, due to a decreased ability to collaborate with providers and others, would have significantly higher HbA(1c) levels than patients with secure attachment style. RESEARCH DESIGN AND METHODS: From 276 adult tertiary care patients with type 1 diabetes, we obtained mean HbA(1c) levels over the prior year and assessed patient attachment style, demographics, and clinical characteristics. We used chi(2) tests and logistic regression to determine whether attachment style was associated with HbA(1c) levels. RESULTS: We found that 62% of patients with dismissing attachment style had mean HbA(1c) levels >or=8% compared with 34% of patients with secure attachment style (P = 0.002). After adjusting for demographics, diabetes severity, medical comorbidity, and depression, dismissing attachment style remained significantly associated with HbA(1c) levels >or=8%, compared with secure attachment style (odds ratio 2.5, 95% CI 1.1-6.0). CONCLUSIONS: We have found that dismissing attachment style is associated with a higher risk for poor glycemic control. This has relevance from a population-based perspective because approximately 25% of the general population has a dismissing attachment style. Attachment style is easily measured using self-report instruments and may inform clinicians how to work with patients who are less engaged in the health care relationship. PMID- 11919134 TI - Morning hyperglycemic excursions: a constant failure in the metabolic control of non-insulin-using patients with type 2 diabetes. AB - OBJECTIVE: To determine whether, over daytime, one or several hyperglycemic excursions exist that can be general failures in the glycemic control of patients with type 2 diabetes. RESEARCH DESIGN AND METHODS: In 200 non-insulin-using patients with type 2 diabetes, diurnal plasma glucose and insulin profiles were studied. Plasma glucose concentrations were measured after an overnight fast (at 8:00 A.M. immediately before breakfast), during the postprandial period (at 11:00 A.M. and 2:00 P.M.), and during the postabsorptive period (at 5:00 P.M., extended postlunch time). RESULTS: In the population considered as a whole, prelunch glucose concentrations (12.0 mmol/l) were found to be significantly increased (P < 0.0001) when compared with those observed at 8:00 A.M. (8.8 mmol/l), at 2:00 P.M. (10.5 mmol/l), and at 5:00 P.M. (8.6 mmol/l). Similar significant excursions (P < 0.0001) in prelunch glucose were observed within subsets of patients selected from the following criteria: 1) body weight, 2) HbA(1c), 3) categories of treatment, and 4) residual beta-cell function. From the calculation of areas under the daytime glucose curves, the relative contributions of postprandial and fasting glucose to the total glucose increment were found to be similar. CONCLUSIONS: High plasma glucose excursions over morning periods seem to be a permanent failure in non-insulin-using patients with type 2 diabetes, whatever the clinical (BMI), biological (HbA(1c)), therapeutic, and pathophysiological (residual beta-cell function) status. Midmorning glucose testing should be recommended for detecting such abnormalities and for correcting them with appropriate therapies. PMID- 11919135 TI - The prevention or delay of type 2 diabetes. PMID- 11919137 TI - Finally, it is our turn! PMID- 11919138 TI - New insights in obesity. PMID- 11919139 TI - Cigarette smoking affects glycemic control in diabetes. PMID- 11919140 TI - Selective screening for gestational diabetes in Chinese women. PMID- 11919141 TI - Beneficial effect of diabetes on acute intermittent porphyria. PMID- 11919142 TI - Risk factors of developing proliferative retinopathy in type 1 diabetic patients : role of BMI. PMID- 11919145 TI - Childhood abuse and diabetes in the community. PMID- 11919144 TI - Comorbidity of type 1 diabetes and anorexia nervosa in a 6-year-old girl. PMID- 11919143 TI - Is type 1 diabetes transmissible by bone marrow allograft? PMID- 11919146 TI - Postischemic microcirculatory blood flow correlates negatively and independently with plasma C-reactive protein in longstanding type 1 diabetes. PMID- 11919147 TI - Two-hour 75-g oral glucose tolerance test early in pregnancy detects most cases of gestational diabetes. PMID- 11919150 TI - Intercellular transfer of antigen-presenting cell determinants onto T cells: molecular mechanisms and biological significance. AB - Upon physiological stimulation, receptors with tyrosine kinase activity (RTK) are rapidly internalized together with their soluble ligands. T cell activation is the consequence of recognition by the T cell receptor (TCR) of specific peptide major histocompatibility protein complexes (peptide-MHC) present at the membrane of antigen-presenting cells (APC). The TCR belongs to the RTK family and is known to be endocytosed upon ligand recognition. It differs from most other RTK in that its ligand, the peptide-MHC complex, is membrane bound and the TCR-ligand interaction is quite weak. Recent experiments have shown that the TCR ligand becomes internalized by T cells upon stimulation. Here we review current knowledge on the molecular mechanisms by which the membrane-bound MHC molecules can be transferred onto T cells, and propose hypotheses on the role this phenomenon could play in physio-pathological situations involving T cells. PMID- 11919151 TI - Cyclase-associated proteins: CAPacity for linking signal transduction and actin polymerization. AB - Many extracellular signals elicit changes in the actin cytoskeleton, which are mediated through an array of signaling proteins and pathways. One family of proteins that plays a role in regulating actin remodeling in response to cellular signals are the cyclase-associated proteins (CAPs). CAPs are highly conserved monomeric actin binding proteins present in a wide range of organisms including yeast, fly, plants, and mammals. The original CAP was isolated as a component of the Saccharomyces cerevisiae adenylyl cyclase complex that serves as an effector of Ras during nutritional signaling. CAPs are multifunctional molecules that contain domains involved in actin binding, adenylyl cyclase association in yeast, SH3 binding, and oligomerization. Genetic studies in yeast have implicated CAPs in vesicle trafficking and endocytosis. CAPs play a developmental role in multicellular organisms, and studies of Drosophila have illuminated the importance of the actin cytoskeleton during eye development and in establishing oocyte polarity. This review will highlight the critical structural and functional domains of CAPs, describe recent studies that have implied important roles for these proteins in linking cell signaling with actin polymerization, and highlight their roles in vesicle trafficking and development. PMID- 11919152 TI - Vascular smooth muscle and nitric oxide synthase. AB - The concept of endothelium-derived relaxing factor (EDRF) put forward in 1980 by Furchgott and Zawadzki implies that nitric oxide (NO) produced by NO synthase (NOS) in the endothelium diffuses to the underlying vascular smooth muscle, where it modulates vascular tone as well as vascular smooth muscle cell (VSMC) proliferation by increasing cGMP formation with subsequent activation of cGMP dependent protein kinase. According to this concept, VSMC do not express NOS by themselves. This attractive, simple scheme is now under considerable debate. To address this issue, we designed this study with the use of a novel supersensitive immunocytochemical technique of signal amplification with tyramide and electron microscopic immunogold labeling complemented with Western blotting, as in our recent studies demonstrating NOS in the myocardial and skeletal muscles. We provide the first evidence that, in contrast to the currently accepted view, VSMC in various blood vessels express all three NOS isoforms depending on the blood vessel type. These findings suggest an alternative mechanism by which local NOS expression may modulate vascular functions in an endothelium-independent manner. PMID- 11919153 TI - Chronic central leptin infusion restores hyperglycemia independent of food intake and insulin level in streptozotocin-induced diabetic rats. AB - We examined the effects of chronic centrally administered leptin on the glucose metabolism of streptozotocin-induced diabetic (STZ-D) rats, a model for insulin dependent diabetes mellitus. When 3 microg.rat(-1).day(-1) of leptin was infused into the third ventricle for 6 consecutive days (STZ-LEP), STZ-D rats became completely euglycemic. The effect was not seen when the same dosage was administered s.c. Centrally administered leptin did not affect peripheral insulin levels. The feeding volume of STZ-LEP rats was suppressed to the level of non-STZ D control rats. No improvement of hyperglycemia was noted when STZ-D rats were pair-fed to match the feeding volume of STZ-LEP rats. Thus, the euglycemia of STZ LEP rats cannot be due to the decreased feeding volume. In the STZ-D rat, glucokinase mRNA, a marker of glycolysis, is down-regulated whereas glucose-6 phosphatase mRNA, a marker of gluconeogenesis, and glucose transporter (GLUT) 2, which is implicated in the release of glucose from liver, are up-regulated. GLUT4, uncoupling protein (UCP) 1, and UCP3 were down-regulated in brown adipose tissue. These parameters returned to normal upon central infusion of leptin. GLUT4 was not down-regulated in the skeletal muscle of STZ-D rats; however, fatty acid binding protein and carnitine palmitoyltransferase I, markers for utilization and beta-oxidation of fatty acids, were up-regulated and restored when the rats were treated with leptin. The increase and subsequent decrease of fatty acid utilization suggests a decrease of glucose uptake in the skeletal muscle of STZ-D rats, which was restored upon central leptin administration. We conclude that centrally infused leptin does not control serum glucose by regulating feeding volume or elevating peripheral insulin, but by regulating hepatic glucose production, peripheral glucose uptake, and energy expenditure. The present study indicates the possibility of future development of a new class of anti-diabetic agents that act centrally and independent of insulin action. PMID- 11919154 TI - Differential membrane targeting of the SERCA and PMCA calcium pumps: experiments with recombinant chimeras. AB - Structural features underlying retention of the SERCA pump in intracellular compartments and the sorting of the PMCA pump to the plasma membrane are not known. The biochemical properties of the two pumps suggest that their differential localization may respond to specific functional demands. The two pumps may control Ca(2+) gradients of different magnitude and dynamic properties. For instance, it has recently become clear that the Ca(2+) gradient across the endoplasmic reticulum (ER) membrane is smaller than that across the plasma membrane. Previous experiments with chimerical constructs of the SERCA and PMCA pumps had suggested a role for the amino-terminal domain in the ER retention of the SERCA pump. Experiments aimed at narrowing down the region responsible for the retention now indicate that the first 28 amino acids of the SERCA pump may play a role in membrane localization. Results also suggest that the formation of oligomers (possibly through the first 28 amino acids) might be critical to the retention mechanism. PMID- 11919155 TI - Activation of an alternative NF-kappaB pathway in skeletal muscle during disuse atrophy. AB - Although cytokine-induced nuclear factor kappaB (NF-kappaB) pathways are involved in muscle wasting subsequent to disease, their potential role in disuse muscle atrophy has not been characterized. Seven days of hind limb unloading led to a 10 fold activation of an NF-kappaB-dependent reporter in rat soleus muscle but not the atrophy-resistant extensor digitorum longus muscle. Nuclear levels of p50 were markedly up-regulated, c-Rel was moderately up-regulated, Rel B was down regulated, and p52 and p65 were unchanged in unloaded solei. The nuclear IkappaB protein Bcl-3 was increased. There was increased binding to an NF-kappaB consensus oligonucleotide, and this complex bound antibodies to p50, c-Rel, and Bcl-3 but not other NF-kappaB family members. Tumor necrosis factor alpha (TNF alpha) and TNF receptor-associated factor 2 protein were moderately down regulated. There was no difference in p38, c-Jun NH(2)-terminal kinase or Akt activity, nor were activator protein 1 or nuclear factor of activated T cell dependent reporters activated. Thus, whereas several NF-kappaB family members are up-regulated, the prototypical markers of cytokine-induced activation of NF kappaB seen with disease-related wasting are not evident during disuse atrophy. Levels of an anti-apoptotic NF-kappaB target, Bcl-2, were increased fourfold whereas proapoptotic proteins Bax and Bak decreased. The evidence presented here suggests that disuse muscle atrophy is associated with activation of an alternative NF-kappaB pathway that involves the activation of p50 but not p65. PMID- 11919156 TI - A new strategy for defining critical functional groups on heparan sulfate. AB - Heparan sulfate (HS) is a sulfated polysaccharide present on cell surfaces and in the extracellular matrix. Accumulating evidence shows that HS plays key roles in many biological systems by interacting with various proteins in a structural specific manner. Due to technical difficulties, however, the understanding of critical functional groups on HS for protein interaction is vague. We report a rapid, convenient, sensitive, and inexpensive strategy using in vitro modification with pure enzymes and gel mobility shift assay to study the subject. We demonstrated the requirements of 3-O, 6-O sulfates and the minimal length of oligosaccharide for antithrombin III (AT-III) binding. We regenerated the binding sites for AT-III on completely desulfated N-resulfated heparin and revealed the critical modification enzymes. This new strategy could be used to identify critical functional groups on HS and to generate HS library and biologically active HS, providing information applicable to the design of HS drugs, such as anticoagulant reagents and viral infection blockers. The binding assay with fibroblast growth factors and receptors confirmed the general usefulness of this approach. PMID- 11919157 TI - HIV-1 Tat protein induces interleukin-10 in human peripheral blood monocytes: involvement of protein kinase C-betaII and -delta. AB - In HIV-infected patients, production of interleukin-10 (IL-10), a highly immunosuppressive cytokine, is associated with the disease progression toward AIDS. We have previously shown that HIV-1 Tat induces IL-10 production by human monocytes via a protein kinase C (PKC) -dependent pathway. Here we show that PKC activation by Tat is essential for IL-10 induction. Among the eight PKC isoforms present in human monocytes, we investigated which isoform(s) plays this crucial role in Tat-mediated IL-10 production and show that 1) Tat can activate PKC alpha, PKC-betaII, PKC-delta, and PKC-epsilon, 2) of these four potential candidates, only PKC-betaII, PKC-delta, and PKC-epsilon are activated by the active domain Tat 1-45, which is responsible for IL-10 production and depleted by long-term exposure to PMA, which abolishes Tat-mediated IL-10 production, 3) whereas selective inhibition of PKC-alpha and PKC-epsilon by specific antisense oligonucleotides has no effect on Tat-mediated IL-10 induction, inhibition of either PKC-betaII or PKC-delta partially inhibits IL-10 production; and 4) the simultaneous inhibition of PKC-betaII and PKC-delta totally inhibits Tat-mediated IL-10. Altogether, these results suggest that the induction of IL-10 by Tat is strictly dependent on the PKC-delta and -betaII isoforms. PMID- 11919158 TI - MT1-MMP expression promotes tumor growth and angiogenesis through an up regulation of vascular endothelial growth factor expression. AB - Membrane type 1 metalloprotease (MT1-MMP) is a transmembrane metalloprotease that plays a major role in the extracellular matrix remodeling, directly by degrading several of its components and indirectly by activating pro-MMP2. We investigated the effects of MT1-MMP overexpression on in vitro and in vivo properties of human breast adenocarcinoma MCF7 cells, which do not express MT1-MMP or MMP-2. MT1-MMP and MMP-2 cDNAs were either transfected alone or cotransfected. All clones overexpressing MT1-MMP 1) were able to activate endogenous or exogenous pro-MMP 2, 2) displayed an enhanced in vitro invasiveness through matrigel-coated filters independent of MMP-2 transfection, 3) induced the rapid development of highly vascularized tumors when injected subcutaneously in nude mice, and 4) promoted blood vessels sprouting in the rat aortic ring assay. These effects were observed in all clones overexpressing MT1-MMP regardless of MMP-2 expression levels, suggesting that the production of MMP-2 by tumor cells themselves does not play a critical role in these events. The angiogenic phenotype of MT1-MMP-producing cells was associated with an up-regulation of VEGF expression. These results emphasize the importance of MT1-MMP during tumor angiogenesis and open new opportunities for the development of anti-angiogenic strategies combining inhibitors of MT1-MMP and VEGF antagonists. PMID- 11919159 TI - RhoA- and RhoD-dependent regulatory switch of Galpha subunit signaling by PAR-1 receptors in cellular invasion. AB - Thrombin and proteinase-activated receptors (PAR) specifically regulate several functions that markedly enhance the transformation phenotype such as inflammation, cell proliferation, tumor growth, and metastasis. We recently reported that thrombin inhibits cellular invasion induced by src, hepatocyte growth factor (HGF), and leptin in kidney and colonic epithelial cells via predominant activation of the pertussis toxin (PTx) -sensitive G-proteins Galphao/Galphai. We provide pharmacological and biochemical evidence that in the presence of PTx, PAR-1 induced cellular invasion through Galpha12/Galpha13- and RhoA/Rho kinase (ROCK) -dependent signaling. However, inhibition of the endogenous small GTPase RhoA by the C3 exoenzyme, dominant-negative N19-RhoA, activated G26V-RhoD, and activators of the nitric oxide/cGMP pathways conferred invasive activity to PAR-1 via a signaling cascade using Galphaq, phospholipase C (PLC), Ca(2+)/calmodulin myosin light chain kinase (CaM-MLCK), and phosphorylation of MLC. We found that cellular invasion induced by the src oncogene is abrogated by inhibitors of the RhoA/ROCK pathway and is independent of PLC/CaM-MLCK signaling. Our data demonstrate that the RhoA and RhoD small GTPases are acting as a molecular switch of cellular invasion and reveal a novel critical mechanism by which PAR-1 bypass Galphao/i and RhoA inhibition via differential coupling to heterotrimeric G-proteins linked to divergent or convergent biological responses. Our data also indicate that Rho GTPases and ROCK mediate a src-dependent invasion signal in kidney and colonic cancer cells. We conclude that dynamic regulation of Rho GTPases activation and inactivation by oncogenes, growth factors, cGMP-inducing agents, and adhesion molecules can initiate convergent invasion signals controlled by the thrombin PAR-1 in cancer cells.-Nguyen, Q.-D., Faivre, S., Bruyneel, E., Rivat, C., Seto, M., Endo, T., Mareel, M., Emami, S., Gespach, C. RhoA- and RhoD-dependent regulatory switch of Galpha subunit signaling by PAR-1 receptors in cellular invasion. PMID- 11919160 TI - Rationale for the role of osteoclast-like cells in arterial calcification. AB - Atherosclerotic arteries frequently become calcified, and these calcium deposits are associated with a high risk of adverse clinical events. Descriptive studies suggest calcification is an organized and regulated process with many similarities to osteogenesis, yet the mechanism and its relationship to atherosclerosis remain largely unknown. In bone development and homeostasis, mineral deposition by osteoblasts and mineral resorption by osteoclasts are delicately balanced such that there is no overall gain or loss in bone mass. We hypothesize that there exists in arteries a mechanism that similarly balances mineral deposition with resorption. We propose that the cellular mediators of arterial mineral resorption are osteoclast-like cells (OLCs) derived from hematopoietic precursors of the mononuclear phagocytic lineage. In arterial microenvironments, mononuclear precursors are induced to differentiate toward OLCs by macrophage-colony stimulating factor and receptor activator of NF-kappaB ligand, both of which are necessary and sufficient for osteoclastogenesis and mineral resorption in bone. OLCs may participate in normal mineral homeostasis within the arterial wall or, alternatively, may be recruited to specific sites within developing plaque. Net calcium deposition occurs as a result of focal perturbation of the balance between the activity of osteoblast-like cells and OLCs. Our proposed mechanism thus views arterial mineral deposition not so much as an active pathological process, but as a localized failure of protective mechanisms that actively oppose mineral deposition within the disordered metabolic milieu of developing atherosclerotic plaque. PMID- 11919161 TI - Role of vasodilator-stimulated phosphoprotein in PKA-induced changes in endothelial junctional permeability. AB - At sites of ongoing inflammation, polymorphonuclear leukocytes (PMN, neutrophils) migrate across vascular endothelia, and such transmigration has the potential to disturb barrier properties and can result in intravascular fluid loss and edema. It was recently appreciated that endogenous pathways exist to dampen barrier disruption during such episodes and may provide an important anti-inflammatory link. For example, during transmigration, PMN-derived adenosine activates endothelial adenosine receptors and induces a cAMP-dependent resealing of endothelial barrier function. In our study reported here, we sought to understand the link between cyclic nucleotide elevation and increased endothelial barrier function. Initial studies revealed that adenosine-induced barrier function is tightly linked to activation of protein kinase A (PKA). Because PKA selectively phosphorylates serine and threonine residues, we screened zonula occludens-1 (ZO 1) immunoprecipitates for the existence of such phosphorylated proteins as targets for barrier regulation. This analysis revealed a dominantly phosphorylated band at 50 kDa. Microsequencing identified this protein as vasodilator-stimulated phosphoprotein (VASP), an actin binding protein with multiple serine/threonine phosphorylation sites. Immunofluorescent microscopy revealed that VASP localizes to endothelial junctional complexes and colocalizes with ZO-1, occludin, and junctional adhesion molecule-1 (JAM-1). To address the role of phospho-VASP in regulation of barrier function, we generated a phosphospecific VASP antibody targeting the Ser157 residue phosphorylation site, the site preferred by PKA. Immunolocalization studies with this antibody revealed that upon PKA activation, phospho-VASP appears at cell-cell junctions. Transient transfection of truncated VASP fragments revealed a parallel increase in barrier function. Taken together, these studies reveal a central role for phospho-VASP in the coordination of PKA-regulated barrier function, such as occurs during episodes of inflammation. PMID- 11919162 TI - NG2 proteoglycan promotes angiogenesis-dependent tumor growth in CNS by sequestering angiostatin. AB - During embryogenesis, the NG2 proteoglycan is expressed on immature capillary vessels, but as the vessels mature they lose this expression. NG2 is up-regulated in high-grade gliomas, but it is not clear to what extent it contributes to malignant progression. Using a combination of high spatial and temporal resolution functional magnetic resonance imaging and histopathological analyses, we show here that overexpression of NG2 increases tumor initiation and growth rates, neovascularization, and cellular proliferation, which predisposes to a poorer survival outcome. By confocal microscopy and cDNA gene array expression profiles, we also show that NG2 tumors express lower levels of hypoxia inducible factor-1a, vascular endothelial growth factor, and endogenous angiostatin in vivo compared with wild-type tumors. Moreover, we demonstrate that NG2-positive cells bind, internalize, and coimmunoprecipitate with angiostatin. These results indicate a unique role for NG2 in regulating the transition from small, poorly vascularized tumors to large, highly vascular gliomas in situ by sequestering angiostatin. PMID- 11919163 TI - Characteristic gene expression profile of primary human cerebral endothelial cells. AB - Endothelial cells of blood vessels forming the interphase between systemic circulation and tissues are crucial for maintenance of homeostasis and organ related functions. Recent experiments support organ-specific endothelial differentiation and suggest differential gene expression patterns in endothelial cells. Here, we compared gene expression in primary human cerebral endothelial cells (HCEC), which are major constituents of the blood brain barrier (BBB), with human umbilical vein endothelial cells (HUVEC) by using cDNA array analysis of 375 genes. Under basal culture conditions, 35 genes were expressed only in HCEC, whereas 20 gene transcripts were detected only in HUVEC. A total of 78 genes were expressed in both endothelial cell types partly with distinct expression levels. Genes expressed by cerebral endothelial cells are important in vasculo- and angiogenesis (VEGF, erbB1) and immunoregulation (OSM-Rbeta, decorin, IL-6) or have growth-supporting properties (brain-derived neurotrophic factor, stem cell factor, transforming growth factor-beta). The differential gene expression profiles were confirmed at the protein level of cell cultures (ELISA, immunoblotting) and human tissues (immunohistochemistry). Identification and further functional characterization of genes specifically expressed by cerebral endothelial cells will have important impact on our understanding of endothelial function at the BBB. PMID- 11919164 TI - The estrogen-regulated protein, TFF1, stimulates migration of human breast cancer cells. AB - The human trefoil protein TFF1 is a small cysteine-rich secreted protein that is frequently expressed in breast tumors under the control of estrogen. The function of TFF1 in breast cancer is unknown. To test the hypothesis that it promotes tumor dissemination, we produced recombinant TFF1 and assessed its ability to stimulate the movement of breast cancer cells by using in vitro wounding and migration assays. Recombinant TFF1 stimulated migration at concentrations of TFF1 found in culture medium. Migration of MCF-7 breast cancer cells, which secrete TFF1, was stimulated by lower concentrations of TFF1 than MDA MB231 cells that do not produce TFF1. Dimeric TFF1, linked by a disulfide bond, and monomeric TFF1 are produced by estrogen-responsive breast cancer cell lines. Recombinant TFF1 dimer was eightfold more potent than TFF1 monomer, implying that the interaction of TFF1 with its receptor is facilitated by dimerization. The majority of TFF1 stimulated migration resulted from chemotaxis, but dimeric TFF1 stimulated some chemokinesis. These results show that estrogens can stimulate the motility of breast cancer cells via the induction of TFF1 and suggest that one reason for the efficacy of hormonal therapies is their ability to reduce expression of TFF1 and, hence, the migration of breast tumor cells. PMID- 11919165 TI - Expression of a catalytically inactive form of diacylglycerol kinase alpha induces sustained signaling through RasGRP. AB - Regulating the generation and clearance of lipid second messengers, such as diacylglycerol (DAG), is critical for the correct propagation of intracellular signaling pathways. DAGK type alpha acts as a negative modulator of the DAG levels generated during T cell activation, which is initiated by triggering of the endogenous T cell receptor (TCR), as well as by stimulation of an ectopically expressed human muscarinic type 1 receptor. Here we show that stimulation of either of these receptors causes rapid, transient membrane translocation of the recently discovered Ras guanyl nucleotide release protein (RasGRP), followed by activation of mitogen-activated protein kinase (MAPK). When cells expressing a catalytically inactive form of DAGKalpha were analyzed, however, similar agonist stimulation resulted in sustained signaling through RasGRP and MAPK. Biochemical analysis showed that expression of kinase-dead diacylglycerol kinase a (DGKalpha) led to a greater, more sustained, DAG accumulation following receptor stimulation. These results suggest that, in T cells, agonist-stimulated DAG generation is the key factor controlling activation of the Ras/MAPK signaling pathway through membrane translocation of RasGRP. Moreover, we demonstrate that through the modulation of the intracellular level of agonist-stimulated DAG, DGKalpha alters Ras activation and downstream signaling dramatically, a process of utmost importance for sound immunological function. PMID- 11919166 TI - Lack of ERK activation and cell migration in FGF-2-deficient endothelial cells. AB - The formation of blood capillaries from preexisting vessels (angiogenesis) and vascular remodeling secondary to atherosclerosis or vessel injury are characterized by endothelial cell migration and proliferation. Numerous growth factors control these cell functions. Basic fibroblast growth factor (FGF-2), a potent angiogenesis inducer, stimulates endothelial cell proliferation, migration, and proteinase production in vitro and in vivo. However, mice genetically deficient in FGF-2 have no apparent vascular defects. We have observed that endothelial cell migration in response to mechanical damage in vitro is accompanied by activation of the extracellular signal-regulated kinase (ERK) pathway, which can be blocked by neutralizing anti-FGF-2 antibodies. Endothelial cells from mice that are genetically deficient in FGF-2 neither migrate nor activate ERK in response to mechanical wounding. Addition of exogenous FGF-2 restores a normal cell response, which shows that impaired migration results from the genetic deficiency of this growth factor. Injury induced ERK activation in endothelial cells occurs only at the edge of the wound. In addition, FGF-2-induced ERK activation mediates endothelial cell migration in response to wounding without a significant effect on proliferation. These data show that FGF-2 is a key regulator of endothelial cell migration during wound repair. PMID- 11919167 TI - Microglial activation and amyloid-beta clearance induced by exogenous heat-shock proteins. AB - Alzheimer's disease (AD) is characterized by the accumulation of fibrillar amyloid-beta (Abeta) peptides to form amyloid plaques. Understanding the balance of production and clearance of Abeta peptides is the key to elucidating amyloid plaque homeostasis. Microglia in the brain, associated with senile plaques, are likely to play a major role in maintaining this balance. Here, we show that heat shock proteins (HSPs), such as HSP90, HSP70, and HSP32, induce the production of interleukin 6 and tumor necrosis factor alpha and increase the phagocytosis and clearance of Abeta peptides. This suggests that microglial interaction with Abeta peptides is highly regulated by HSPs. The mechanism of microglial activation by exogenous HSPs involves the nuclear factor kB and p38 mitogen-activated protein kinase pathways mediated by Toll-like receptor 4 activation. In AD brains, levels of HSP90 were increased in both the cytosolic and membranous fractions, and HSP90 was colocalized with amyloid plaques. These observations suggest that HSP-induced microglial activation may serve a neuroprotective role by facilitating Abeta clearance and cytokine production PMID- 11919168 TI - Simulated microgravity alters differentiation and increases apoptosis in human follicular thyroid carcinoma cells. AB - This study focuses on the effects of simulated microgravity (0g) on the human follicular thyroid carcinoma cell line ML-1. Cultured on a three-dimensional clinostat, ML-1 cells formed three-dimensional MCTSs (MCTS diameter: 0.3 +/- 0.01 mm). After 24 and 48 h of clinorotation, the cells significantly decreased fT3 and fT4 secretion but up-regulated the thyroid-stimulating hormone-receptor expression as well as the production of vimentin, vinculin, and extracellular matrix proteins (collagen I and III, laminin, fibronectin, chondroitin sulfate) compared with controls. Furthermore, ML-1 cells grown on the clinostat showed elevated amounts of the apoptosis-associated Fas protein, of p53, and of bax but showed reduced quantities of bcl-2. In addition, signs of apoptosis became detectable, as assessed by terminal deoxynucleotidyl transferase-mediated dUTP digoxigenin nick end labeling, 4', 6-diamidino-2-phenylindole staining, DNA laddering, and 85-kDa apoptosis-related cleavage fragments. These fragments resulted from enhanced 116-kDa poly(ADP-ribose)polymerase (PARP) activity and apoptosis. These observations suggest that clinorotation elevates intermediate filaments, cell adhesion molecules, and extracellular matrix proteins and simultaneously induces apoptosis in follicular thyroid cancer cells. In conclusion, our experiments could provide a regulatory basis for the finding that astronauts show low thyroid hormone levels after space flight, which may be explained by the increase of apoptosis in thyrocytes as a result of simulated 0g. PMID- 11919170 TI - Acute sodium deficit triggers plasticity of the brain angiotensin type 1 receptors. AB - The brain renin-angiotensin system (bRAS) is involved in the control of hydromineral balance. However, little information is available on the functional regulation of the bRAS as a consequence of sodium deficit in the extracellular fluid compartments. We used a pharmacological model of acute Na+ depletion (furosemide injections) to investigate changes of a major component of the bRAS, the hypothalamic angiotensin type 1A (AT(1A)) receptors. Furosemide induced a rapid and long-lasting expression of the AT(1A) mRNA in the subfornical organ, the median preoptic nucleus (MnPO), and the parvocellular division of the paraventricular nucleus (pPVN). Na+ depletion increased the number of cells expressing AT(1A) mRNA in the pPVN, but not in the MnPO. The enhancement of AT(1A) mRNA expression was associated with an increase in AT(1) binding sites in all the regions studied. It is of interest that in the paraventricular nucleus, the majority of the neurons expressing AT(1A) mRNA also showed an increase in metabolic activity (Fos-related antigen immunoreactivity [FRA-ir]). By contrast, in the MnPO, we observe two distinct cell populations. Our data demonstrated that an acute Na+ deficit induced a functional regulation of the hypothalamic AT(1A) receptors, indicating that these receptors are subject to plasticity in response to hydromineral perturbations. PMID- 11919169 TI - The permeability transition pore signals apoptosis by directing Bax translocation and multimerization. AB - Mitochondria are key players of apoptosis and can irreversibly commit the cell to death by releasing cytochrome c (Cyt.c) to the cytosol, where caspases 9 and 3 subsequently get activated. Under conditions of oxidative stress, opening of the mitochondrial permeability transition pore (PTP) represents an early trigger and is crucial in causing Cyt.c release. To account for the latter, current models propose that PTP gating would result, as is the case in vitro, in the rupture of the outer mitochondrial membrane caused by mitochondrial matrix swelling. Using live cell imaging and recombinant fluorescent probes based on the green fluorescent protein (GFP) and its mutants, we report that directed repetitive gating of the PTP triggers a delayed Cyt.c efflux, which is not associated with mitochondrial swelling. Instead, subcellular imaging shows that PTP opening signals the redistribution of the cytosolic protein Bax to the mitochondria, where it secondarily forms clusters that appear to be a prerequisite for Cyt.c release. Fluorescence resonance energy transfer imaging further reveals that Bax clustering coincides with the formation of Bax multimers. We conclude that the PTP is not itself a component of the Cyt.c release machinery, but that it acts indirectly by signaling Bax translocation and multimerization. PMID- 11919171 TI - An increase in the myocardial PCr/ATP ratio in GLUT4 null mice. AB - ATP and creatine phosphate (PCr) are prime myocardial high-energy phosphates. Their relative concentrations are conserved among mammalian species and across a range of physiologic cardiac workloads. The cardiac PCr/ATP ratio is decreased with several pathologic conditions, such as ischemia and heart failure, but there are no reports of an increase in the cardiac PCr/ATP ratio in any species or with interventions. We studied the in vivo energetics in transgenic mice lacking expression of the glucose transport protein GLUT4 (G4N) and observed a significant 60% increase in the myocardial PCr/ATP ratio in G4N that was confirmed in three different experimental settings including intact animals. The higher PCr/ATP in G4N is cardiac-specific and is due to higher total cardiac creatine (CR) concentrations in G4N than in wild-type (WT). However, [ATP], [ADP], and -DG(-ATP) did not differ between the strains. Expression of the creatine transport protein (CreaT) that is responsible for creatine uptake in myocytes was preserved in G4N cardiac tissue. These observations demonstrate, for the first time to our knowledge, that G4N manifest a unique increase in the cardiac PCr/ATP ratio, which suggests a novel genetic strategy for increasing myocardial creatine levels. PMID- 11919172 TI - HIV enhances substance P expression in human immune cells. AB - Substance P (SP), a potent modulator of neuroimmunoregulation, is expressed in human immune cells. We observed elevated plasma SP levels in HIV-infected men compared with uninfected subjects. In the present study, we investigated the possible cellular source of the increased SP level caused by HIV infection. Using real-time reverse transcriptase-polymerase chain reaction, we demonstrated that monocyte-derived macrophages (MDM) and lymphocytes from both placental cord blood and adult peripheral blood expressed SP mRNA, which was significantly increased by HIV infection. HIV-induced SP expression was positively related to virus replication in the infected MDM. Purified recombinant HIV envelope glycoprotein 120 (gp120) derived from both the macrophage-tropic strain (MN) and the T lymphocyte-tropic strain (IIIB), when added to MDM cultures, enhanced SP mRNA expression. The gp120-induced SP expression was abrogated by pretreating the cells with soluble CD4. Furthermore, the activation of HIV in the latently infected promonocytic cell line (U1) and T-cell line (ACH-2) up-regulated SP mRNA expression. These data support the hypothesis that interaction of HIV and SP may have significant in vivo relevance to the immunopathogenesis of HIV infection and AIDS. PMID- 11919173 TI - Perturbations in choline metabolism cause neural tube defects in mouse embryos in vitro. AB - A role for choline during early stages of mammalian embryogenesis has not been established, although recent studies show that inhibitors of choline uptake and metabolism, 2-dimethylaminoethanol (DMAE), and 1-O-octadecyl-2-O-methyl-rac glycero-3-phosphocholine (ET-18-OCH3), produce neural tube defects in mouse embryos grown in vitro. To determine potential mechanisms responsible for these abnormalities, choline metabolism in the presence or absence of these inhibitors was evaluated in cultured, neurulating mouse embryos by using chromatographic techniques. Results showed that 90%-95% of 14C-choline was incorporated into phosphocholine and phosphatidylcholine (PtdCho), which was metabolized to sphingomyelin. Choline was oxidized to betaine, and betaine homocysteine methyltransferase was expressed. Acetylcholine was synthesized in yolk sacs, but 70 kDa choline acetyltransferase was undetectable by immunoblot. DMAE reduced embryonic choline uptake and inhibited phosphocholine, PtdCho, phosphatidylethanolamine (PtdEtn), and sphingomyelin synthesis. ET-18-OCH3 also inhibited PtdCho synthesis. In embryos and yolk sacs incubated with 3H ethanolamine, 95% of recovered label was PtdEtn, but PtdEtn was not converted to PtdCho, which suggested that phosphatidylethanolamine methyltransferase (PeMT) activity was absent. In ET-18-OCH3 treated yolk sacs, PtdEtn was increased, but PtdCho was still not generated through PeMT. Results suggest that endogenous PtdCho synthesis is important during neurulation and that perturbed choline metabolism contributes to neural tube defects produced by DMAE and ET-18-OCH3. PMID- 11919174 TI - Altered mechanical properties and intracellular calcium signaling in cardiomyocytes from annexin 6 null-mutant mice. AB - Annexin 6 is one of a widely expressed family of calcium-binding proteins found in most mammalian tissues, including the heart. Several studies have implicated annexin 6 in the regulation of intracellular Ca2+ signaling, and it has been shown in vitro to act as a modulator of the sarcoplasmic reticulum Ca2+-release channel, cardiac L-type calcium channel, and Na+/Ca2+ exchanger. To investigate the role of annexin 6 in intact cardiomyocytes, we used mice containing a targeted disruption of the annexin 6 gene. Compared with controls, the myocytes of annexin 6 null-mutant mice demonstrated a significant increase in the rates of shortening and relengthening. Intracellular Ca2+ transients in fura-2-loaded cardiomyocytes induced by caffeine showed a normal baseline and amplitude, whereas the rate of decay was doubled in annexin 6-/- myocytes compared with control mice. These results show that annexin 6 knockout in the mouse leads to an increase in myocyte contractility and faster diastolic Ca2+ removal from the cytoplasm. In light of published findings showing annexin 6 to be down-regulated in end-stage heart failure, these results are consistent with a role for annexin 6 as a negative inotropic factor in the regulation of cardiomyocyte mechanics. PMID- 11919175 TI - Sphingosine 1-phosphate induces chemotaxis of immature and modulates cytokine release in mature human dendritic cells for emergence of Th2 immune responses. AB - Sphingosine 1-phosphate (S1P) is a potent extracellular lysolipid phosphoric acid mediator that is released after IgE-stimulation of mast cells. Here we investigated the biological activity and intracellular signaling of S1P on human dendritic cells (DC), which are specialized antigen presenting cells with the ability to migrate into peripheral tissues and lymph nodes, as well as control the activation of naive T cells. We show that immature and mature DC express the mRNA for different S1P receptors, such as endothelial differentiation gene (EDG) 1, EDG-3, EDG-5, and EDG-6. In immature DC, S1P stimulated pertussis toxin sensitive Ca2+ increase actin-polymerization and chemotaxis. These responses were lost by DC matured with lipopolysaccharide. In maturing DC, however, S1P inhibited the secretion of tumor necrosis factor alpha and interleukin (IL)-12, whereas it enhanced secretion of IL-10. As a consequence, mature DC exposed to S1P showed a reduced and increased capacity to generate allogeneic Th1 and Th2 responses, respectively. In summary, our study implicates that S1P might regulate the trafficking of DC and ultimately favor Th2 lymphocyte-dominated immunity. PMID- 11919176 TI - Myocardial proteome analysis reveals reduced NOS inhibition and enhanced glycolytic capacity in areas of low local blood flow. AB - In the heart, in situ local myocardial blood flow (MBF) varies greater than 10 fold between individual areas and displays a spatially heterogeneous pattern. To analyze its molecular basis, we analyzed protein expression of low and high flow samples (300 mg, <50% or >150% of mean MBF, each n=30) of six beagle dogs by 2-D polyacrylamide gel electrophoresis (380 +/- 78 spots/gel). In low flow samples, dimethylarginine dimethylaminohydrolase (DDAH1) was increased greatly (+377%, compared with high flow samples). This increase resulted in a 75% reduction of asymmetric dimethylarginine (ADMA), the potent endogenous inhibitor of NO synthase, whereas eNOS showed no difference. Low flow samples exhibited enhanced expression of GAPDH (+89%) and phosphoglycerate kinase (+100%), whereas hydroxyacyl-CoA dehydrogenase, electron transfer flavoprotein, myoglobin, and desmin were decreased. Assessing local MBF on different days within 2 weeks revealed a high degree of MBF stability (r2 > 0.79). Thus, stable differences in local MBF are associated with significant differences in local gene and protein expression. In low flow areas, the increased DDAH1 reduces ADMA concentration and NOS inhibition, which strongly suggests enhanced NO formation. Low flow areas are also characterized by a higher glycolytic and a lower fatty acid oxidation capacity. Both the shift in substrate utilization and the rise in NO may contribute to the known lower oxygen consumption in these areas. PMID- 11919177 TI - A repressor in the proximal human inducible nitric oxide synthase promoter modulates transcriptional activation. AB - The human inducible nitric oxide synthase (iNOS or NOSII) gene is regulated through an extended and complex promoter. In this study, the transcriptional regulation of human NOSII is investigated in the human colon cell line HCT-8R. Stimulation with a cytokine mix (interferon-gamma, interleukin 1-beta, and tumor necrosis factor alpha) induces NOSII mRNA accumulation, as well as promoter activity in these cells. Several random deletions were performed within the proximal 7 kb of the promoter, which led to the identification of a region, whose deletion provokes a marked increase in transcriptional activity upon cytokine stimulation. Furthermore, this region is shown to repress a viral-driven luciferase construct, mainly at basal levels. An AP-1-like sequence present in this region that is specifically recognized by nuclear proteins is shown to be involved in the repressive effect. This element is capable of repressing a viral promoter, and its deletion augments cytokine-stimulated transcription. These findings are confirmed in various cell lines and suggest a general mechanism for the control of basal levels of NOSII expression, to avoid unnecessary toxicity under normal conditions. PMID- 11919178 TI - Nucleomorphin. A novel, acidic, nuclear calmodulin-binding protein from dictyostelium that regulates nuclear number. AB - Probing of Dictyostelium discoideum cell extracts after SDS-PAGE using (35)S recombinant calmodulin (CaM) as a probe has revealed approximately three-dozen Ca(2+)-dependent calmodulin binding proteins. Here, we report the molecular cloning, expression, and subcellular localization of a gene encoding a novel calmodulin-binding protein (CaMBP); we have called nucleomorphin, from D. discoideum. A lambdaZAP cDNA expression library of cells from multicellular development was screened using a recombinant calmodulin probe ((35)S-VU1-CaM). The open reading frame of 1119 nucleotides encodes a polypeptide of 340 amino acids with a calculated molecular mass of 38.7 kDa and is constitutively expressed throughout the Dictyostelium life cycle. Nucleomorphin contains a highly acidic glutamic/aspartic acid inverted repeat (DEED) with significant similarity to the conserved nucleoplasmin domain and a putative transmembrane domain in the carboxyl-terminal region. Southern blotting reveals that nucleomorphin exists as a single copy gene. Using gel overlay assays and CaM agarose we show that bacterially expressed nucleomorphin binds to bovine CaM in a Ca(2+)-dependent manner. Amino-terminal fusion to the green fluorescence protein (GFP) showed that GFP-NumA localized to the nucleus as distinct arc-like patterns similar to heterochromatin regions. GFP-NumA lacking the acidic DEED repeat still showed arc-like accumulations at the nuclear periphery, but the number of nuclei in these cells was increased markedly compared with control cells. Cells expressing GFP-NumA lacking the transmembrane domain localized to the nuclear periphery but did not affect nuclear number or gross morphology. Nucleomorphin is the first nuclear CaMBP to be identified in Dictyostelium. Furthermore, these data present the first identification of a member of the nucleoplasmin family as a calmodulin-binding protein and suggest nucleomorphin has a role in nuclear structure in Dictyostelium. PMID- 11919180 TI - Kidney sulfatides in mouse models of inherited glycosphingolipid disorders: determination by nano-electrospray ionization tandem mass spectrometry. AB - Sulfatides show structural, and possibly physiological similarities to gangliosides. Kidney dysfunction might be correlated with changes in sulfatides, the major acidic glycosphingolipids in this organ. To elucidate their in vivo metabolic pathway these compounds were analyzed in mice afflicted with inherited glycosphingolipid disorders. The mice under study lacked the genes encoding either beta-hexosaminidase alpha-subunit (Hexa-/-), the beta-hexosaminidase beta subunit (Hexb-/-), both beta-hexosaminidase alpha and beta-subunits (Hexa-/- and Hexb-/-), GD3 synthase (GD3S-/-), GD3 synthase and GalNAc transferase (GD3S-/- and GalNAcT-/-), GM2 activator protein (Gm2a-/-), or arylsulfatase A (ASA-/-). Quantification of the sulfatides, I(3)SO(3)(-)-GalCer (SM4s), II(3)SO(3)(-) LacCer (SM3), II(3)SO(3)(-)-Gg(3)Cer (SM2a), and IV(3,) II(3)-(SO(3)(-))(2) Gg(4)Cer (SB1a), was performed by nano-electrospray tandem mass spectrometry. We conclude for the in vivo situation in mouse kidneys that: 1) a single enzyme (GalNAc transferase) is responsible for the synthesis of SM2a and GM2 from SM3 and GM3, respectively. 2) In analogy to GD1a, SB1a is degraded via SM2a. 3) SM2a is hydrolyzed to SM3 by beta-hexosaminidase S (Hex S) and Hex A, but not Hex B. Both enzymes are supported by GM2-activator protein. 4) Arylsulfatase A is required to degrade SB1a. It is probably the sole sphingolipid-sulfatase cleaving the galactosyl-3-sulfate bond. In addition, a human Tay-Sachs patient's liver was investigated, which showed accumulation of SM2a along with GM2 storage. The different ceramide compositions of both compounds indicated they were probably derived from different cell types. These data demonstrate that in vivo the sulfatides of the ganglio-series follow the same metabolic pathways as the gangliosides with the replacement of sulfotransferases and sulfatases by sialyltransferases and sialidases. Furthermore, a novel neutral GSL, IV(6)GlcNAcbeta-Gb(4)Cer, was found to accumulate only in Hexa-/- and Hexb-/- mouse kidneys. From this we conclude that Hex S also efficiently cleaves terminal beta1-6-linked HexNAc residues from neutral GSLs in vivo. PMID- 11919179 TI - The anti-HIV pentameric pseudopeptide HB-19 binds the C-terminal end of nucleolin and prevents anchorage of virus particles in the plasma membrane of target cells. AB - The multivalent pseudopeptide HB-19 that binds the cell-surface-expressed nucleolin is a potent inhibitor of human immunodeficiency virus (HIV) infection by blocking virus particle attachment and thus anchorage in the plasma membrane. We show that cross-linking of surface-bound HB-19A (like HB-19 but with a modified template) results in aggregation of HB-19A with surface nucleolin. Consistent with its specific action, HB-19A binding to different types of cells reaches saturation at concentrations that have been reported to result in inhibition of HIV infection. By using Chinese hamster ovary mutant cell lines, we confirm that the binding of HB-19A to surface nucleolin is independent of heparan and chondroitin sulfate proteoglycans. In vitro generated full-length nucleolin was found to bind HB-19A, whereas the N-terminal part containing the acidic amino acid stretches of nucleolin did not. The use of various deletion constructs of the C-terminal part of nucleolin then permitted the identification of the extreme C-terminal end of nucleolin, containing repeats of the amino acid motif, RGG, as the domain that binds HB-19A. Finally, a synthetic peptide corresponding to the last C-terminal 63 amino acids was able to inhibit HIV infection at the stage of HIV attachment to cells, thus suggesting that this domain could be functional in the HIV anchorage process. PMID- 11919181 TI - Dose-dependent activation of antiapoptotic and proapoptotic pathways by ethanol treatment in human vascular endothelial cells: differential involvement of adenosine. AB - Moderate but not heavy drinking has been found to have a protective effect against cardiovascular morbidity. We investigated the effects of ethanol (EtOH) treatment on the cell survival-promoting phosphatidylinositol 3-kinase (PI3K)/Akt pathway in cultured human umbilical vein endothelial cells (HUVEC). Exposure of cells to 2-20 mm EtOH resulted in rapid (<10 min) induction of Akt phosphorylation that could be prevented by pertussis toxin or the PI3K inhibitors wortmannin and LY294002. Among the downstream effectors of PI3K/Akt, p70S6 kinase, glycogen synthase kinase 3alpha/beta, and IkappaB-alpha were phosphorylated, the latter resulting in 3-fold activation of NF-kappaB. EtOH also activated p44/42 mitogen-activated protein kinase in a PI3K-dependent manner. Low concentrations of EtOH increased endothelial nitric-oxide synthase activity, which could be blocked by transfection of HUVEC with dominant-negative Akt, implicating the PI3K/Akt pathway in this effect. The adenosine A1 receptor antagonist 1,3-dipopylcyclopentylxanthine prevented the phosphorylation of Akt observed in the presence of EtOH, adenosine, or the A1 agonist N(6) cyclopentyladenosine. Incubation of HUVEC with 50-100 mm EtOH resulted in mitochondrial permeability transition and caspase-3 activation followed by apoptosis, as documented by DNA fragmentation and TUNEL assays. EtOH-induced apoptosis was unaffected by DPCPX and was potentiated by wortmannin or LY294002. We conclude that treatment with low concentrations of EtOH activates the cell survival promoting PI3K/Akt pathway in endothelial cells by an adenosine receptor dependent mechanism and activation of the proapoptotic caspase pathway by higher concentrations of EtOH via an adenosine-independent mechanism can mask or counteract such effects. PMID- 11919182 TI - A fragment of paxillin binds the alpha 4 integrin cytoplasmic domain (tail) and selectively inhibits alpha 4-mediated cell migration. AB - The alpha(4) integrins play important roles in embryogenesis, hematopoiesis, cardiac development, and the immune responses. The alpha(4) integrin subunit is indispensable for these biological processes, possibly because the alpha(4) subunit regulates cellular functions differently from other integrin alpha subunits. We have previously reported that the alpha(4) cytoplasmic domain directly and tightly binds paxillin, an intracellular signaling adaptor molecule, and this interaction accounts for some of the unusual functional responses to alpha(4) integrin-mediated cell adhesion. We also have identified a conserved 9 amino acid region (Glu(983)-Tyr(991)) in the alpha(4) cytoplasmic domain that is sufficient for paxillin binding, and an alanine substitution at either Glu(983) or Tyr(991) within this region disrupted the alpha(4)-paxillin interaction and reversed the effects of the alpha(4) cytoplasmic domain on cell spreading and migration. In the current study, we have mapped the alpha(4)-binding site within paxillin using mutational analysis, and examined its effects on the alpha(4) tail mediated functional responses. Here we report that sequences between residues Ala(176) and Asp(275) of paxillin are sufficient for binding to the alpha(4) tail. We found that the alpha(4) tail, paxillin, and FAT, the focal adhesion targeting domain of pp125(FAK), could form a ternary complex and that the alpha(4)-binding paxillin fragment, P(Ala(176)-Asp(275)), specifically blocked paxillin binding to the alpha(4) tail more efficiently than it blocked binding to FAT. Furthermore, when expressed in cells, this alpha(4)-binding paxillin fragment specifically inhibited the alpha(4) tail-stimulated cell migration. Thus, paxillin binding to the alpha(4) tail leads to enhanced cell migration and inhibition of the alpha(4)-paxillin interaction selectively blocks the alpha4 dependent cellular responses. PMID- 11919183 TI - Identification of essential residues in the type II Hsp40 Sis1 that function in polypeptide binding. AB - Sis1 is an essential yeast Type II Hsp40 protein that assists cytosolic Hsp70 Ssa1 in the facilitation of processes that include translation initiation, the prevention of protein aggregation, and proteasomal protein degradation. An essential function of Sis1 and other Hsp40 proteins is the binding and delivery of non-native polypeptides to Hsp70. How Hsp40s function as molecular chaperones is unknown. The crystal structure of a Sis1 fragment that retains peptide-binding activity suggests that Type II Hsp40s utilize hydrophobic residues located in a solvent-exposed patch on carboxyl-terminal domain I to bind non-native polypeptides. To test this model, amino acid residues Val-184, Leu-186, Lys-199, Phe-201, Ile-203, and Phe-251, which form a depression in carboxyl-terminal domain I, were mutated, and the ability of Sis1 mutants to support cell viability and function as molecular chaperones was examined. We report that Lys-199, Phe 201, and Phe-251 are essential for cell viability and required for Sis1 polypeptide binding activity. Sis1 I203T could support normal cell growth, but when purified it exhibited severe defects in chaperone function. These data identify essential residues in Sis1 that function in polypeptide binding and help define the nature of the polypeptide-binding site in Type II Hsp40 proteins. PMID- 11919184 TI - Structure/function relationships in OxlT, the oxalate/formate antiporter of Oxalobacter formigenes: assignment of transmembrane helix 2 to the translocation pathway. AB - We constructed a single cysteine panel encompassing transmembrane helix two (TM2) of OxlT, the oxalate/formate antiporter of Oxalobacter formigenes. Among the 21 positions targeted, cysteine substitution identified one (phenylalanine 59) as essential to OxlT expression and three (glutamine 56, glutamine 66, and serine 69) as potentially critical to OxlT function. By probing membranes with a bulky hydrophilic probe (Oregon Green maleimide) we also located a central inaccessible core of at least eight residues in length, extending from leucine 61 to glycine 68. Functional assays based on reconstitution of crude detergent extracts showed that of single cysteine mutants within the TM2 core only the Q63C variant was substantially (> or =95%) inhibited by thiol-specific agents (carboxyethyl methanethiosulfonate and ethylsulfonate methanethiosulfonate). Subsequent analytical work using the purified Q63C protein showed that inhibition by ethylsulfonate methanethiosulfonate was blocked by substrate and that the concentration dependence of such substrate protection occurred with a binding constant of 0.16 mm oxalate, comparable with the Michaelis constant observed for oxalate transport (0.23 mm). These findings lead us to conclude that position 63 lies on the OxlT translocation pathway. Our conclusion is strengthened by the finding that position 63, along with most other positions relevant to TM2 function, is found on a helical face that can be cross-linked to the pathway facing surface of TM11 (Fu, D., Sarker, R. I., Bolton, E., and Maloney, P. C. (2001) J. Biol. Chem. 276, 8753-8760). PMID- 11919185 TI - A ligand-inducible epidermal growth factor receptor/anaplastic lymphoma kinase chimera promotes mitogenesis and transforming properties in 3T3 cells. AB - Oncogenic rearrangements of the anaplastic lymphoma kinase (ALK) gene, encoding a receptor type tyrosine kinase, are frequently associated with anaplastic large cell lymphomas. Such rearrangements juxtapose the intracellular domain of ALK to 5'-end sequences belonging to different genes and create transforming fusion proteins. To understand how the oncogenic versions of ALK contribute to lymphomagenesis, it is important to analyze the biological effects and the biochemical properties of this receptor under controlled conditions of activation. To this aim, we constructed chimeric receptor molecules in which the extracellular domain of the ALK kinase is replaced by the extracellular, ligand binding domain of the epidermal growth factor receptor (EGFR). Upon transfection in NIH 3T3 fibroblasts, the EGFR/ALK chimera was correctly synthesized and transported to the cell surface, where it was fully functional in forming high versus low affinity EGF-binding sites and transducing an EGF-dependent signal intracellularly. Overexpression of the EGFR/ALK chimera in NIH 3T3 was sufficient to induce the malignant phenotype; the appearance of the transformed phenotype was, however, conditionally dependent on the administration of EGF. Moreover, the EGFR/ALK chimera was significantly more active in inducing transformation and DNA synthesis than the wild type EGFR when either was expressed at similar levels in NIH 3T3 cells. Comparative analysis of the biochemical pathways implicated in the transduction of mitogenic signals did not show any increased ability of the EGFR/ALK to phosphorylate PLC-gamma and MAPK compared with the EGFR. On the contrary, EGFR/ALK showed to have a consistently greater effect on phosphatidylinositol 3-kinase activity compared with the EGFR, indicating that this enzyme plays a major role in mediating the mitogenic effects of ALK in NIH 3T3 cells. PMID- 11919186 TI - The estrogen-responsive B box protein: a novel regulator of keratinocyte differentiation. AB - Keratinocyte growth factor (KGF) regulates proliferation, differentiation, migration, and survival of different types of epithelial cells, including keratinocytes of the skin. To gain insight into the mechanisms underlying these multiple functions, we searched for KGF- regulated genes in keratinocytes. Using the differential display reverse transcriptase-PCR technology, we identified the gene encoding the estrogen-responsive B box protein (EBBP) which has as yet not been functionally characterized. The full-length murine and human EBBP cDNAs were cloned and fully sequenced. They were shown to encode 75-kDa proteins, which are mainly localized in the cytoplasm of keratinocytes in vitro and in vivo. In vivo, EBBP was found at high levels in the KGF- and epidermal growth factor-responsive basal keratinocytes of human skin, but the expression was down-regulated in the hyperthickened epithelium of skin wounds. Stable overexpression of EBBP in HaCaT keratinocytes did not affect the proliferation rate of the transfected cells, but enhanced the early differentiation process. These results suggest that the presence of EBBP in basal keratinocytes is important for the differentiation capacity of these cells, and that down-regulation of EBBP expression in a hyperproliferative epithelium is required to maintain the cells in a non differentiated stage. PMID- 11919187 TI - In vitro selection of RNA aptamers that bind to cell adhesion receptors of Trypanosoma cruzi and inhibit cell invasion. AB - Trypanosoma cruzi causing Chagas' disease needs to invade host cells to complete its life cycle. Macromolecules on host cell surfaces such as laminin, thrombospondin, heparan sulfate, and fibronectin are believed to be important in mediating parasite-host cell adhesions and in the invasion process of the host cell by the parasite. The SELEX technique (systematic evolution of ligands by exponential enrichment) was used to evolve nuclease-resistant RNA ligands (aptamer = to fit) that bind with affinities of 40-400 nm to parasite receptors for the host cell matrix molecules laminin, fibronectin, thrombospondin, and heparan sulfate. After eight consecutive rounds of in vitro selection four classes of RNA aptamers based on structural similarities were isolated and sequenced. All members of each class shared a common sequence motif and competed with the respective host cell matrix molecule that was used for displacement during the selection procedure. RNA pools following seven and eight selection rounds as well as individual aptamers sharing consensus motifs were active in inhibiting invasion of LLC-MK(2) monkey kidney cells by T. cruzi in vitro. PMID- 11919188 TI - A Forkhead/winged helix-related transcription factor mediates insulin-increased plasminogen activator inhibitor-1 gene transcription. AB - Plasminogen activator inhibitor-1 (PAI-1) is an important regulator of fibrinolysis by its inhibition of both tissue-type and urokinase plasminogen activators. PAI-1 levels are elevated in type II diabetes and this elevation correlates with macro- and microvascular complications of diabetes. Insulin increases PAI-1 production in several experimental systems, but the mechanism of insulin-activated PAI-1 transcription remains to be determined. Deletion analysis of the PAI-1 promoter revealed that the insulin response element is between -117 and -7. Mutation of the AT-rich site at -52/-45 abolished the insulin responsiveness of the PAI-1 promoter. This sequence is similar to the inhibitory sequence found in the phosphoenolpyruvate carboxylkinase/insulin-like growth factor-I-binding protein I promoters. Gel-mobility shift assays demonstrated that the forkhead bound to the PAI-1 promoter insulin response element. Expression of the DNA-binding domain of FKHR acted as a dominant negative to block insulin increased PAI-1-CAT expression. A LexA-FKHR construct was also insulin responsive. These data suggested that a member of the Forkhead/winged helix family of transcription factors mediated the effect of insulin on PAI-1 transcription. Inhibition of phosphatidylinositol 3-kinase reduced the effect of insulin on PAI-1 gene expression, a result consistent with activation through FKHR. However, it was likely that a different member of the FKHR family (not FKHR) mediated this effect since FKHR was present in both insulin-responsive and non-responsive cell lines. PMID- 11919189 TI - Integrin cytoplasmic domain-associated protein 1alpha (ICAP-1alpha ) interacts directly with the metastasis suppressor nm23-H2, and both proteins are targeted to newly formed cell adhesion sites upon integrin engagement. AB - Cell adhesion-dependent signaling implicates cytoplasmic proteins interacting with the intracellular tails of integrins. Among those, the integrin cytoplasmic domain-associated protein 1alpha (ICAP-1alpha) has been shown to interact specifically with the beta(1) integrin cytoplasmic domain. Although it is likely that this protein plays an important role in controlling cell adhesion and migration, little is known about its actual function. To search for potential ICAP-1alpha-binding proteins, we used a yeast two-hybrid screen and identified the human metastatic suppressor protein nm23-H2 as a new partner of ICAP-1alpha. This direct interaction was confirmed in vitro, using purified recombinant ICAP 1alpha and nm23-H2, and by co-immunoprecipitation from CHO cell lysates over expressing ICAP-1alpha. The physiological relevance of this interaction is provided by confocal fluorescence microscopy, which shows that ICAP-1alpha and nm23-H2 are co-localized in lamellipodia during the early stages of cell spreading. These adhesion sites are enriched in occupied beta(1) integrins and precede the formation of focal adhesions devoid of ICAP-1alpha and nm23-H2, indicating the dynamic segregation of components of matrix adhesions. This peripheral staining of ICAP-1alpha and nm23-H2 is only observed in cells spreading on fibronectin and collagen and is absent in cells spreading on poly-l lysine, vitronectin, or laminin. This is consistent with the fact that targeting of both ICAP-1alpha and nm23-H2 to the cell periphery is dependent on beta(1) integrin engagement rather than being a consequence of cell adhesion. This finding represents the first evidence that the tumor suppressor nm23-H2 could act on beta(1) integrin-mediated cell adhesion by interacting with one of the integrin partners, ICAP-1alpha. PMID- 11919190 TI - Ets1 is an effector of the transforming growth factor beta (TGF-beta ) signaling pathway and an antagonist of the profibrotic effects of TGF-beta. AB - Extracellular matrix (ECM) production and turnover are tightly controlled under normal physiological conditions. Ets factors regulate matrix turnover by activating transcription of several metalloproteinases (MMPs) and are frequently overexpressed in aggressive tumors and arthritis. Because of the prominent role of transforming growth factor beta (TGF-beta) in ECM synthesis, this study was undertaken to determine the possible interactions between Ets1 and the TGF-beta pathway. Experiments using adenoviral delivery of Ets1 in human fibroblasts have established that Ets1 strongly suppresses TGF-beta induction of collagen type I and other matrix-related genes and reverses TGF-beta-dependent inhibition of MMP 1. Subsequent experiments utilizing COL1A2 promoter demonstrated that Ets1 in the presence of TGF-beta signaling interferes with the stimulatory role of p300. To gain further insight into the mechanism of Ets1 inhibition of the TGF-beta signaling, the protein levels and post-translational modifications of Ets1 after TGF-beta treatment were analyzed. The level of total Ets1 protein was not affected after 24 h of TGF-beta stimulation. Moreover, TGF-beta did not affect either serine or threonine phosphorylation levels of Ets1. However, TGF-beta induced rapid and prolonged lysine acetylation of Ets1. In addition, analyses of endogenous p300.Ets1 complexes revealed that acetylated Ets1 is preferentially associated with the p300/CBP complexes. TGF-beta treatment leads to dissociation of Ets1 from the CBP/p300 complexes. Together, these findings suggest that elevated expression of Ets1 in fibroblasts fundamentally alters their responses to TGF-beta in favor of matrix degradation and away from matrix deposition as exemplified by arthritis and cancer. PMID- 11919191 TI - The 7472insC mitochondrial DNA mutation impairs the synthesis and extent of aminoacylation of tRNASer(UCN) but not its structure or rate of turnover. AB - The 7472insC mitochondrial DNA mutation in the tRNA(Ser(UCN)) gene is associated with sensorineural deafness combined, in some patients, with a wider neurological syndrome. In cultured cybrid cells it causes a 70% decrease in tRNA(Ser(UCN)) abundance and mild respiratory impairment, previously suggested to be due to decreased tRNA stability. When mitochondrial transcription was blocked by ethidium bromide treatment, the half-life of the mutant tRNA was not significantly different from that of wild-type tRNA(Ser(UCN)). Over-expression of mitochondrial translational elongation factor EF-Tu also had no effect on the mutant phenotype. However, during recovery from prolonged ethidium bromide treatment, the synthesis of the mutant tRNA(Ser(UCN)) was specifically impaired, without polarity effects on downstream tRNAs of the light strand transcription unit. We infer that the mutation acts posttranscriptionally to decrease tRNA(Ser(UCN)) abundance by affecting its synthesis rather than its stability. The extent of aminoacylation of the mutant tRNA was also decreased by approximately 25%. In contrast, the mutation had no detectable effect on tRNA(Ser(UCN)) base modification or structure other than the insertion of an extra guanosine templated by the mutation, which was structurally protected from nuclease digestion like the surrounding nucleotides. These findings indicate a common molecular process underlying sensorineural deafness caused by mitochondrial tRNA(Ser(UCN)) mutations. PMID- 11919192 TI - Activation of caspase-9 is required for UV-induced apoptosis of human keratinocytes. AB - UV radiation from the sun activates both the membrane death receptor and the intrinsic or mitochondrial apoptotic signaling pathways in epidermal keratinocytes, triggering apoptosis and affording protection against skin cancer formation. We have investigated the involvement of caspase-9 in the UV death effector pathway in human keratinocytes, since this is the initiating caspase in the mitochondrial pathway required for UV-induced apoptosis in some, but not all, cell types. UV radiation triggered activation of caspase-3, caspase-9, and caspase-8 with similar kinetics, although the rank order of activation was caspase-3 > caspase-9 > caspase-8. Inhibition of caspase-9 with either the peptide inhibitor benzyloxycarbonyl-Leu-Glu(OCH(3))-His-Asp(OCH(3))-fluoromethyl ketone, or expression of a catalytically inactive caspase-9 by retroviral transduction, protected normal keratinocytes from UV-induced apoptosis. HaCaT keratinocytes harboring mutant p53 alleles were also protected from UV-induced apoptosis by the dominant negative caspase-9. The dominant negative caspase-9 blocked UV-induced activation of caspase-3, caspase-9, and caspase-8, and also protected cells from the loss of mitochondrial membrane potential. In contrast, the dominant negative caspase-9 did not protect from anti-Fas-induced apoptosis or caspase activation. These results identify caspase-9 as the critical upstream caspase initiating apoptosis by UV radiation in human keratinocytes, the relevant cell type for this important environmental carcinogen. PMID- 11919193 TI - Ku is important for telomere maintenance, but not for differential expression of telomeric VSG genes, in African trypanosomes. AB - Trypanosome antigenic variation, involving differential expression of variant surface glycoprotein (VSG) genes, has a strong association with telomeres and with DNA recombination. All expressed VSGs are telomeric, and differential activation involves recombination into the telomeric environment or silencing/activation of subtelomeric promoters. A number of pathogen contingency gene systems associated with immune evasion involve telomeric loci, which has prompted speculation that chromosome ends provide conditions conducive for the operation of rapid gene switching mechanisms. Ku is a protein associated with eukaryotic telomeres that is directly involved in DNA recombination and in gene silencing. We have tested the hypothesis that Ku in trypanosomes is centrally involved in differential VSG expression. We show, via the generation of null mutants, that trypanosome Ku is closely involved in telomere length maintenance, more so for a transcriptionally active than an inactive telomere, but exhibits no detectable influence on DNA double strand break repair. The absence of Ku and the consequent great shortening of telomeres had no detectable influence either on the rate of VSG switching or on the silencing of the telomeric promoters of the VSG subset that is expressed in the tsetse fly. PMID- 11919194 TI - Colocalization, physical, and functional interaction between Werner and Bloom syndrome proteins. AB - The RecQ helicase family comprises a conserved group of proteins implicated in several aspects of DNA metabolism. Three of the family members are defective in heritable diseases characterized by abnormal growth, premature aging, and predisposition to malignancies. These include the WRN and BLM gene products that are defective in Werner and Bloom syndromes, disorders which share many phenotypic and cellular characteristics including spontaneous genomic instability. Here, we report a physical and functional interaction between BLM and WRN. These proteins were coimmunoprecipitated from a nuclear matrix solubilized fraction, and the purified recombinant proteins were shown to interact directly. Moreover, BLM and WRN colocalized to nuclear foci in three human cell lines. Two regions of WRN that mediate interaction with BLM were identified, and one of these was localized to the exonuclease domain of WRN. Functionally, BLM inhibited the exonuclease activity of WRN. This is the first demonstration of a physical and functional interaction between RecQ helicases. Our observation that RecQ family members interact provides new insights into the complex phenotypic manifestations resulting from the loss of these proteins. PMID- 11919195 TI - Phosphatase inhibition leads to histone deacetylases 1 and 2 phosphorylation and disruption of corepressor interactions. AB - The regulation of histone deacetylases (HDACs) by phosphorylation was examined by elevating intracellular phosphorylation in cultured cells with the protein phosphatase inhibitor okadaic acid. After fractionation of extracts from treated versus untreated cells, HDAC 1 and 2 eluted in several peaks of deacetylase activity, assayed using mixed acetylated histones or acetylated histone H4 peptide. Stimulation of cells with okadaic acid led to hyperphosphorylation of HDAC 1 and 2 as well as changes in column elution of both enzymes. Hyperphosphorylated HDAC2 was also observed in cells synchronized with nocodazole or taxol, demonstrating regulation of HDAC phosphorylation during mitosis. Phosphorylated HDAC1 and 2 showed a gel mobility retardation that correlated with a small but significant increase in activity, both of which were reversed upon phosphatase treatment in vitro. However, the most pronounced effect of HDAC phosphorylation was to disrupt protein complex formation between HDAC1 and 2 as well as complex formation between HDAC1 and corepressors mSin3A and YY1. In contrast, interactions between HDAC1/2 and RbAp46/48 were unaffected by okadaic acid. These results establish a novel link between HDAC phosphorylation and the control of protein-protein interactions and suggest a mechanism for relief of deacetylase-catalyzed transcriptional repression by phosphorylation-dependent signaling. PMID- 11919196 TI - Cyclin E is a target of WT1 transcriptional repression. AB - WT1 was originally identified as a Wilms' tumor suppressor gene, but it may have oncogenic potential in leukemia and in some solid tumors. WT1 is a transcription factor that has been implicated in the regulation of target genes related to apoptosis, genitourinary differentiation, and cell cycle progression. Because induction of WT1 leads indirectly to increased p21 expression in osteosarcoma cells, we investigated the possibility that other genes involved in the G(1)/S phase transition might also be WT1 targets. Cyclin E plays a crucial role in the cell cycle by activating cyclin-dependent kinase 2, which phosphorylates Rb, leading to progression from G(1) into S phase. We identified several WT1 binding sites in the cyclin E promoter. We demonstrate that WT1 binds to these sites and that in transient transfection assays WT1 represses the cyclin E promoter. This activity is dependent on the presence of a binding site located downstream of the transcription start site. In intact cells, induction of WT1 expression down regulates cyclin E protein levels. These results provide the first demonstration that WT1 can directly modulate the expression of a gene involved in cell cycle progression. PMID- 11919197 TI - The effect of cytosine methylation on the structure and geometry of the Holliday junction: the structure of d(CCGGTACm5CGG) at 1.5 A resolution. AB - The single crystal structure of the methylated sequence d(CCGGTACm(5)CGG) has been solved as an antiparallel stacked X Holliday junction to 1.5 A resolution. When compared with the parent nonmethylated d(CCGGTACCGG) structure, the duplexes are translated by 3.4 A along the helix axis and rotated by 10.8 degrees relative to each other, rendering the major grooves more accessible overall. A Ca(2+) complex is seen in the minor groove opposite the junction but is related to the B conformation of the stacked arms. At the junction itself, the hydrogen bond from the N4 nitrogen of cytosine C8 to the C7 phosphate at the crossover in the parent structure has been replaced by a water bridge. Thus, this direct interaction is not absolutely required to stabilize the junction at the previously defined ACC trinucleotide core. The more compact methylated junction forces the Na(+) of the protected central cavity of the nonmethylated junction into a solvent cluster that spans the space between the junction crossover and the stacked arms. A series of void volumes within the methylated and the nonmethylated structures suggests that small monovalent cations can fill and vacate this central cavity without the need to unfold the four-stranded Holliday junction completely. PMID- 11919198 TI - Intermediary glucan structures formed during starch granule biosynthesis are enriched in short side chains, a dynamic pulse labeling approach. AB - The formation of intermediary glucans, mature starch, and phytoglycogen was studied using leaves of Arabidopsis thaliana wild type and dbe mutant, which lacks plastidic isoamylase (Zeeman, S. C., Umemoto, T., Lue, W. L., Au-Yeung, P., Martin, C., Smith, A. M., and Chen, J. (1998) Plant Cell 10, 1699-1711). A new approach to the study of starch biosynthesis was developed based on "very short pulse" labeling of leaf starch through photosynthetic fixation of (14)CO(2). This allowed selective analysis of the structure of starch formed within a 30-s period. This time frame is shorter than the period required for the formation of a single crystalline amylopectin lamella and consequently permits a direct analysis of intermediary structures during granule formation. Analysis of chain length distribution showed that the most recently formed outer layer of the granules has a structure different from the mature starch. The outer layer is enriched in short chains that are 6-11 glucose residues long. Side chains with 6 glucose residues are the shortest abundant chains formed, and they are formed exclusively by transfer from donor chains of 12 glucose residues or longer. The labeling pattern shows that chain transfer resulting in branching is a rapid and efficient process, and the preferential labeling of shorter chains in the intermediary granule bound glucan is suggested to be a direct consequence of efficient branching. Although similar, the short chain intermediary structure is not identical to phytoglycogen, which is an even more highly branched molecule with very few longer chains (more than 40 glucose residues). Pulse and chase labeling profiles for the dbe mutant showed that the final structure is more highly branched than the intermediary structures, which implies that branching of phytoglycogen occurs over a longer time period than branching of starch. PMID- 11919199 TI - Low fidelity DNA synthesis by a y family DNA polymerase due to misalignment in the active site. AB - Sulfolobus solfataricus DNA polymerase IV (Dpo4) is a member of the Y family of DNA polymerases whose crystal structure has recently been solved. As a model for other evolutionarily conserved Y family members that perform translesion DNA synthesis and have low fidelity, we describe here the base substitution and frameshift fidelity of DNA synthesis by Dpo4. Dpo4 generates all 12 base-base mismatches at high rates, 11 of which are similar to those of its human homolog, DNA polymerase kappa. This result is consistent with the Dpo4 structure, implying lower geometric selection for correct base pairs. Surprisingly, Dpo4 generates C.dCMP mismatches at an unusually high average rate and preferentially at cytosine flanked by 5'-template guanine. Dpo4 also has very low frameshift fidelity and frequently generates deletions of even noniterated nucleotides, especially cytosine flanked by a 5'-template guanine. Both unusual features of error specificity suggest that Dpo4 can incorporate dNTP precursors when two template nucleotides are present in the active site binding pocket. These results have implications for mutagenesis resulting from DNA synthesis by Y family polymerases. PMID- 11919200 TI - Biochemical defects in retina-specific human ATP binding cassette transporter nucleotide binding domain 1 mutants associated with macular degeneration. AB - The retina-specific human ABC transporter (ABCR) functions in the retinal transport system and has been implicated in several inherited visual diseases, including Stargardt disease, fundus flavimaculatus, cone-rod dystrophy, and age related macular degeneration. We have previously described a general ribonucleotidase activity of the first nucleotide binding domain (NBD1) of human ABCR (Biswas, E. E. (2001) Biochemistry 40, 8181-8187). In this communication, we present a quantitative study analyzing the effects of certain disease-associated mutations, Gly-863 --> Ala, Pro-940 --> Arg, and Arg-943 --> Gln on the nucleotide binding, and general ribonucleotidase activities of this domain. NBD1 proteins, harboring these mutations, were created through in vitro site-specific mutagenesis and expressed in Escherichia coli. Results of the enzyme-kinetic studies indicated that these mutations altered the ATPase and CTPase activities of NBD1. The G863A and P940R mutations were found to have significant attenuation of the rates of nucleotide hydrolysis and binding affinities. On the other hand, the R943Q mutation had small, but detectable reduction in its nucleotidase activity and nucleotide binding affinity. We have measured the nucleotide binding affinities of NBD1 protein and its mutants quantitatively by fluorescence anisotropy changes during protein binding to ethenoadenosine ATP (epsilonATP), a fluorescent ATP analogue. We have correlated the dissociation constant (K(D)) and the rates of nucleotide hydrolysis (V(max)) of NBD1 and its mutants with the available genetic data for these mutations. PMID- 11919202 TI - Molecular enzymology of the catalytic domains of the Dnmt3a and Dnmt3b DNA methyltransferases. AB - The C-terminal domains of the mammalian DNA methyltransferases Dnmt1, Dnmt3a, and Dnmt3b harbor all the conserved motifs characteristic for cytosine-C5 methyltransferases. Whereas the isolated catalytic domain of Dnmt1 is inactive, we show here that the C-terminal domains of Dnmt3a and Dnmt3b are catalytically active. Neither Dnmt3a nor Dnmt3b shows a significant preference for the satellite 2 sequence, although Dnmt3b is required for methylation of these regions in vivo. However, the catalytic domain of Dnmt3a methylates DNA in a distributive reaction, whereas Dnmt3b is processive, which accelerates methylation of macromolecular DNA in vitro. This property could make Dnmt3b a preferred enzyme for methylation at satellite 2 repeats, since they are highly CG rich. We have also analyzed the catalytic activities of six different mutations found in ICF (immunodeficiency, centromeric instability, and facial abnormalities) patients in the catalytic domain of Dnmt3b. Five of them display catalytic activities reduced by 10-50-fold; one mutant was inactive in our assay (residual activity <1%). These results confirm that a reduced catalytic activity of Dnm3b causes ICF. However, the mutations in general do not completely abrogate catalytic activity. This finding may explain why ICF patients are viable, whereas nmt3b knock-out mice die during embryogenesis. PMID- 11919201 TI - The macrophage C-type lectin specific for galactose/N-acetylgalactosamine is an endocytic receptor expressed on monocyte-derived immature dendritic cells. AB - Lectins on antigen presenting cells are potentially involved in the antigen uptake and the cellular recognition and trafficking. Serial analysis of gene expression in monocyte-derived dendritic cells (DCs), monocytes, and macrophages revealed that 7 of the 19 C-type lectin mRNA were present in immature DCs. Two of these, the macrophage mannose receptor and the macrophage lectin specific for galactose/N-acetylgalactosamine (MGL), were found only in immature DCs, as confirmed by reverse transcriptase-PCR and flow cytometric analysis. By subcloning and sequencing the amplified mRNA, we obtained nucleotide sequences encoding seven different human MGL (hMGL) subtypes, which were apparently derived from alternatively spliced mRNA. In addition, the hMGL gene locus on human chromosome 17p13 contains one gene. A single nucleotide polymorphism was identified at a position in exon 3 that corresponds to the cytoplasmic region proximal to the transmembrane domain. Of all the splicing variants, the hMGL variant 6C was expressed at the highest levels on immature DCs from all donors tested. Immature DCs could incorporate alpha-GalNAc-modified soluble acrylamide polymers, and this was significantly inhibited by pretreatment of the cells with an anti-hMGL monoclonal antibody that blocks the lectin-carbohydrate interaction. We propose that hMGL is a marker of imDCs and that it functions as an endocytic receptor for glycosylated antigens. PMID- 11919203 TI - NMR analysis of bovine tRNATrp: conformation dependence of Mg2+ binding. AB - NMR was used to study the solution structure of bovine tRNA(Trp) hyperexpressed in Escherichia coli. With the use of (15)N labeling and site-directed mutagenesis to assign overlapping resonances through the base pair replacement of U(71)A(2) by G(2)C(71), U(27)A(43) by G(27)C(43), and G(12)C(23) by U(12)A(23), the resonances of all 26 observable imino protons in the helical regions and in the tertiary interactions were assigned unambiguously by means of two-dimensional nuclear Overhauser effect spectroscopy and heteronuclear single quantum coherence methods. When the discriminator base A(73) and the G(12)C(23) base pair on the D stem, two identity elements on bovine tRNA(Trp) that are important for effective recognition by tryptophanyl-tRNA synthetase, were mutated to the ineffective forms of G(73) and U(12)A(23), respectively, NMR analysis revealed an important conformational change in the U(12)A(23) mutant but not in the G(73) mutant molecule. Thus A(73) appears to be directly recognized by tryptophanyl-tRNA synthetase, and G(12)C(23) represents an important structural determinant. Mg(2+) effects on the assigned resonances of imino protons allowed the identification of strong, medium, and weak Mg(2+) binding sites in tRNA(Trp). Strong Mg(2+) binding modes were associated with the residues G(7), s(4)U(8) (where s(4)U is 4 thiouridine), G(12), and U(52). The observations that G(42) was associated with strong Mg(2+) binding in only the U(12)A(23) mutant tRNA(Trp) but not the wild type or G(73) mutant tRNA(Trp) and that the G(7), s(4)U(8), G(24), and G(22) imino protons are associated with a two-site Mg(2+) binding mode in wild type and G(73) mutant but only a one-site mode in the U(12)A(23) mutant established the occurrence of conformational change in the U(12)A(23) mutant tRNA(Trp). These observations also established the dependence of Mg(2+) binding on tRNA conformation and the usefulness of Mg(2+) binding sites as conformational probes. The thermal titration of tRNA(Trp) in the presence and absence of 10 mm Mg(2+) indicated that overall tRNA(Trp) structure stability was increased by more than 15 degrees C by the presence of Mg(2+). PMID- 11919204 TI - Critical features of a conserved RNA stem-loop important for feedback regulation of RNase E synthesis. AB - RNase E is an important regulatory enzyme that governs the principal pathway for mRNA degradation in Escherichia coli. This endonuclease controls its own synthesis via a feedback mechanism in which the longevity of rne (RNase E) mRNA is modulated by a cis-acting sensory element that responds to changes in cellular RNase E activity. Previous research has shown that this element is an RNA stem loop (hp2) within the 5'-untranslated region of the rne transcript. Here we report studies involving mutational analysis and phylogenetic comparison that have identified the features of rne hp2 important for its function. These comprise an internal loop flanked on one side by a 2-bp stem and a hairpin loop and on the other side by a longer stem whose sequence is inconsequential. A search of bacterial genome sequences suggests that regulation by an hp2-like element may be a unique evolutionary adaptation of the rne transcript that is not shared by other mRNAs. PMID- 11919205 TI - Coupling endoplasmic reticulum stress to the cell death program. An Apaf-1 independent intrinsic pathway. AB - Accumulation of misfolded proteins and alterations in Ca2+ homeostasis in the endoplasmic reticulum (ER) causes ER stress and leads to cell death. However, the signal-transducing events that connect ER stress to cell death pathways are incompletely understood. To discern the pathway by which ER stress-induced cell death proceeds, we performed studies on Apaf-1(-/-) (null) fibroblasts that are known to be relatively resistant to apoptotic insults that induce the intrinsic apoptotic pathway. While these cells were resistant to cell death initiated by proapoptotic stimuli such as tamoxifen, they were susceptible to apoptosis induced by thapsigargin and brefeldin-A, both of which induce ER stress. This pathway was inhibited by catalytic mutants of caspase-12 and caspase-9 and by a peptide inhibitor of caspase-9 but not by caspase-8 inhibitors. Cleavage of caspases and poly(ADP-ribose) polymerase was observed in cell-free extracts lacking cytochrome c that were isolated from thapsigargin or brefeldin-treated cells. To define the molecular requirements for this Apaf-1 and cytochrome c independent apoptosis pathway further, we developed a cell-free system of ER stress-induced apoptosis; the addition of microsomes prepared from ER stress induced cells to a normal cell extract lacking mitochondria or cytochrome c resulted in processing of caspases. Immunodepletion experiments suggested that caspase-12 was one of the microsomal components required to activate downstream caspases. Thus, ER stress-induced programmed cell death defines a novel, mitochondrial and Apaf-1-independent, intrinsic apoptotic pathway. PMID- 11919206 TI - Cervical cancer. PMID- 11919207 TI - Paul Langerhans. PMID- 11919210 TI - Increased stool sampling during a waterborne outbreak of cryptosporidiosis does not increase the detection of other faecal pathogens. AB - AIMS: To ascertain the effect of enhanced surveillance following an outbreak of waterborne cryptosporidiosis on the number of faecal specimens submitted to the local microbiology laboratory and the number positive for common enteric pathogens. The outbreak provided an opportunity to estimate the extent of routine under ascertainment of common enteric pathogens. METHOD: Retrospective search of the computerised microbiology system database for details of faecal examination requests for the period 26 April to 6 June in 1998 and 1999 (period of outbreak). RESULTS: Specimens were received from 378 community patients during the six week period 26 April to 6 June 1999. This was double that for the same period in 1998 (a non-outbreak year). Oocysts of Cryptosporidium parvum were detected in 59 patients, an eightfold increase compared with the same period in 1998. Despite the greater number of patients tested, the detection of other pathogens in patients with gastroenteritis was not altered when compared with the same period in the previous year. CONCLUSION: This study found no evidence of under ascertainment of gastrointestinal infection (common bacterial pathogens and rotavirus) by local general practitioners. PMID- 11919209 TI - Current management of hereditary angio-oedema (C'1 esterase inhibitor deficiency). AB - Hereditary angio-oedema is characterised by recurrent swellings in any part of the body and also by recurrent attacks of severe abdominal pain. The disease is inherited in an autosomal dominant manner but up to 25% of cases can occur as a spontaneous mutation. Attacks of swelling can be precipitated by trauma, certain drugs, and emotional stress. Treatment usually involves a combination of prophylaxis, using androgens or antifibrolytic drugs, and replacement with C'1 esterase inhibitor concentrate for acute attacks and before surgery or other traumatic procedures. PMID- 11919211 TI - Death certification: an audit of practice entering the 21st century. AB - AIMS: Death certification, a legal duty of doctors, continues to be poorly performed despite Royal College recommendations and increased education at an undergraduate level. Therefore, the current performance of certifying doctors was audited within a large teaching hospital entering the new century. METHODS: A total of 1000 completed certificate counterfoils were examined retrospectively for appropriateness of completion and the ability to construct a logical cause of death cascade. RESULTS: Only 55% of certificates were completed to a minimally accepted standard, and many of these failed to provide relevant information to allow adequate ICD-10 coding. Nearly 10% were completed to a poor standard, being illogical or inappropriately completed. CONCLUSIONS: The results show no improvement in the state of certification. Possible interventions to improve outcomes are discussed; however, in light of a recent high profile legal case a current Home Office review of death certification may suggest the passing of statutory law to ensure accurate completion. PMID- 11919212 TI - Lipoprotein (a) does not participate in the early acute phase response to training or extreme physical activity and is unlikely to enhance any associated immediate cardiovascular risk. AB - AIMS: To investigate the proposal that lipoprotein (a) (Lp(a)) contributes to the acute phase response and thus possibly to the acute cardiac risks associated with major physical effort. METHODS/RESULTS: Fit, healthy, British army recruits were reviewed at the beginning and the end of a 10 week programme of basic training concluding with an intense 48 hour military exercise. Final recruit assessment was staggered over the last week of training, giving rise to six recruit groups, with determination of Lp(a), C reactive protein (CRP), fibrinogen, albumin, and total creatine kinase values from 12 hours to five days after the final exercise. A clear acute phase response was seen following the final exercise, marked by a significant increase in circulating concentrations of fibrinogen and a reduction of albumin, and a trend with non-significant increases in CRP. CONCLUSION: Lp(a) did not behave as an early marker of the acute response. Previous reports may have been confounded by concurrent disease in older subjects and by late sampling. Lp(a) determination for cardiovascular risk profiling is not confounded by associated physical effort. It is also unlikely that the acute risks of major physical effort are enhanced by any process involving Lp(a). PMID- 11919214 TI - An evaluation of the performance of XLD, DCA, MLCB, and ABC agars as direct plating media for the isolation of Salmonella enterica from faeces. AB - AIMS: To compare the performance of four media, singly and in combination, as direct plating media for the isolation of Salmonella enterica from human faeces. METHODS: Two thousand four hundred and nine routine, faecal samples received by four laboratories were inoculated on to xylose lysine desoxycholate (XLD), desoxycholate citrate (DCA), mannitol lysine crystal violet brilliant green (MLCB), and alpha-beta chromogenic (ABC) agars using standardised protocols, reagents, and data collection. Isolates of presumptive salmonellae were identified using standard laboratory techniques and the results were analysed statistically. RESULTS: Direct plating recovered 46 of the 60 possible isolates of Salmonella spp recovered via enrichment broth. No isolates were recovered from direct plating that were not recovered via selenite enrichment. MLCB gave the highest isolation rate individually (84.8%) and amounts of competing flora (CF) did not affect the recognition of colonies. ABC proved highly specific, but insensitive, and isolation rates were adversely affected by any amount of CF. Isolation rates from XLD and DCA were only affected when the CF load was heavy. DCA was least specific, with only 9.01% of picks positive and greatest number of confirmatory tests. XLD and MLCB, in combination, gave the highest isolation rate. CONCLUSIONS: Where the earlier results of direct plating may be advantageous, XLD and MLCB provide the optimal combination. For non-typhi salmonellae, MLCB is the best, single direct plating medium. For routine diagnostic work, XLD is most effective. PMID- 11919208 TI - The causal relation between human papillomavirus and cervical cancer. AB - The causal role of human papillomavirus infections in cervical cancer has been documented beyond reasonable doubt. The association is present in virtually all cervical cancer cases worldwide. It is the right time for medical societies and public health regulators to consider this evidence and to define its preventive and clinical implications. A comprehensive review of key studies and results is presented. PMID- 11919215 TI - Neonatal alloimmune thrombocytopenia in the Irish population: a discrepancy between observed and expected cases. AB - AIMS: To estimate the rate of detection of neonatal alloimmune thrombocytopenia (NAITP) in the Irish population, to investigate clinical presentation and outcome in affected infants, and to determine the extent, if any, to which this condition is underdiagnosed at present. METHODS: Cases were collected in a retrospective fashion from a review of platelet serology laboratory records from January 1992 to December 2000. Clinical data were obtained from hospital records. Testing for maternal antiplatelet antibody was by one or more of the following: the platelet suspension immunofluorescence test, a commercial antigen capture enzyme linked immunosorbent assay (GTI-PakPlus), and the monoclonal antibody immobilisation of platelet antigens assay. Platelet antigen typing was by the polymerase chain reaction technique with sequence specific primers. RESULTS: Twenty seven serologically verified cases of NAITP were identified in 18 families. Maternal antibody to human platelet antigen 1a accounted for 25 of the 27 confirmed cases. Twenty one of 26 infants were born with severe thrombocytopenia. Nineteen of 27 infants had bleeding manifestations at birth. Petechiae and bruising were most commonly observed (n = 17). There were no documented cases of intracranial haemorrhage in this group but systematic cranial ultrasound was not performed. CONCLUSIONS: Screening studies in predominantly white populations have estimated the incidence of NAITP to be between 1 in 1000 and 1 in 2000 live births. With 50 000 births each year in Ireland, these results give a clinical detection rate for NAITP of just 1 case in 16 500 live births, strongly suggesting that NAITP is currently underdiagnosed. Antenatal screening to detect women at risk of having babies with NAITP is now scientifically feasible and should be considered. PMID- 11919216 TI - Tumour histological grade may progress between primary and recurrent invasive mammary carcinoma. AB - AIMS: To assess the constancy of the histological grade of invasive breast carcinomas by comparing primary tumours with their axillary metastases and local or regional recurrences. METHODS: Eighty four recurrent invasive breast carcinomas with a primary tumour or previous recurrence were available for histological review from the period 1980 to 2000. These and any further recurrences were graded by one observer. RESULTS: Nine, 24, and 51 tumours with grades 1, 2, and 3, respectively, recurred. Grade 1, 2, and 3 tumours recurred within a median time of 88, 42, and 23 months, respectively. The intraobserver reproducibility of the histological grade was good (kappa = 0.66), and the grades of the primary tumours and their axillary metastases or next recurrence also exhibited good agreement. However, when further (second to sixth) recurrences were included in the analysis, the agreement between the grade of the tumours and their last recurrence was only moderate (kappa = 0.48). Only two of the nine grade 1 and 15 of the 24 grade 2 tumours retained their grade in their last recurrence. CONCLUSIONS: Low grade carcinomas require a longer follow up. These long term data support the possibility of a transition from low grade invasive breast carcinomas to higher grade tumours. It is suggested that low grade (well differentiated) breast carcinomas are not a single entity: some may progress to high grade tumours, whereas others appear not to progress. PMID- 11919217 TI - Expression of antimicrobial neutrophil defensins in epithelial cells of active inflammatory bowel disease mucosa. AB - BACKGROUND/AIMS: The normal intestinal epithelium is increasingly being recognised as an important component of the mucosal innate protection against microorganisms. Human neutrophil defensins 1-3 (HNP 1-3) and lysozyme are components of the systemic innate immunity. The aim of this study was to investigate the expression of HNP 1-3 and lysozyme in normal and active inflammatory bowel disease (IBD) mucosa. METHODS: Mucosal tissue sections were studied by immunohistochemistry using antibodies to neutrophil defensins 1-3 and lysozyme. Extracts of purified intestinal epithelial cells were used for immunoblotting studies and antimicrobial activity against the phoP negative strain of Salmonella typhimurium. RESULTS: Surface epithelial cells strongly immunoreactive for neutrophil defensins and lysozyme were seen in active ulcerative colitis and Crohn's disease (but not normal or inactive IBD) mucosal samples. Many of these cells coexpressed both of the antimicrobial proteins. Immunoblotting studies confirmed the expression of neutrophil defensins in extracts of purified ulcerative colitis epithelial cells, which also demonstrated antimicrobial activity. CONCLUSION: HNP 1-3 and lysozyme are expressed in surface enterocytes of mucosa with active IBD and they may play an important role in intestinal host defence against luminal microorganisms. PMID- 11919219 TI - Misidentification of a genomovar of Burkholderia cepacia by recA restriction fragment length polymorphism. AB - An 8 year old girl with cystic fibrosis presented with a pulmonary exacerbation from which Burkholderia cepacia was cultured. Subsequent polymerase chain reaction restriction fragment length polymorphism analysis of the recA gene suggested the presence of B cepacia Genomovar V (Burkholderia vietnamiensis); however, on subsequent sequence typing, this isolate was confirmed as B cepacia Genomovar IIIb. This report outlines the potential difficulties in the correct characterisation of the various genomovars within the B cepacia complex of organisms, which has particularly important implications for patient segregation and infection control. PMID- 11919218 TI - Offline telepathology diagnosis of colorectal polyps: a study of interobserver agreement and comparison with glass slide diagnoses. AB - BACKGROUND/AIMS: Technological advances have produced telepathology systems with high quality colour images and reasonable transmission times. Most applications of telepathology have centred on the remote diagnosis of frozen sections or remote real time expert opinions. This study investigates the reproducibility and accuracy of offline telepathology as a primary diagnostic medium for routine histopathology specimens. METHODS: One hundred colorectal polyps (50 hyperplastic, 50 adenomatous) were presented in a randomised order to five histopathologists as offline images on a telepathology workstation. Six images of each case were used: the slide label, a low power scan of all material on the slide, and four higher magnification views. The times taken to prepare the images, and to make the diagnoses, were recorded. Interobserver agreement was measured with kappa statistics and compared with the glass slide diagnoses. RESULTS: The kappa statistics for the interobserver agreement on the telepathology images lay in the range of 0.90-1.00, which is interpreted as excellent agreement, and were significantly higher than those for the glass slide diagnoses (range, 0.84-0.98; p = 0.001). The median time taken to capture the images for a case was 210 seconds. The median time taken to make a diagnosis from the telepathology images was five seconds, which was significantly shorter than for the glass slide diagnoses (median, 13 seconds; p < 0.0005). CONCLUSIONS: Offline telepathology has the potential to be a primary diagnostic medium for routine histopathology with a high degree of reproducibility and short diagnosis times. Further studies are required to validate offline telepathology for different types of specimens and different operators of the image capture system. PMID- 11919220 TI - Is clinical practice variability the major reason for differences in pathology requesting patterns in general practice? AB - AIMS: To examine whether variations in pathology test requesting between different general practices can be accounted for by sociodemographic or other descriptive indicators of the practice. METHOD: This was a comparative analysis of requesting patterns across a range of pathology tests representing 95% of those requested in general practice, in 22 general practices in a single district, serving a population of 165 000. Spearman correlation coefficients were calculated and both the top and bottom fifths of activity were displayed graphically to detect trends at the extremes of the ranges. RESULTS: The proportion of women of childbearing age, median practice Townsend scores, or the existence of specialist miniclinics within the practice did not have a demonstrable impact on requesting patterns. A weak correlation was found between the proportion of elderly patients and creatinine/electrolyte testing but not for the other two tests examined for this patient group. CONCLUSIONS: The large differences observed in general practice pathology requesting probably result mostly from individual variation in clinical practice and are therefore potentially amenable to change. PMID- 11919221 TI - Reliability of the tissue microarray based FISH for evaluation of the HER-2 oncogene in breast carcinoma. AB - AIMS: Tumour tissue microarray allows the analysis of hundreds of tumour samples simultaneously on a single microscope slide. However, the extremely small tissue samples taken from the original tissue may not always be representative of the entire tumour. METHODS: The reliability of this new technology was investigated by analysing HER-2 oncogene amplification by fluorescence in situ hybridisation (FISH) from representative slides of the whole tumour and small tissue core biopsies from 29 invasive breast tumours. RESULTS: The tissue microarray method had high accuracy; in only one of 29 cases (3.4%; 95% confidence interval, 0% to 10%) were the results discordant with whole tumour analysis. CONCLUSION: Tumour microarray is a highly reliable method for analysing HER-2 oncogene amplification by FISH in human breast tumours. PMID- 11919222 TI - A primary tumour of the oesophagus with both melanocytic and schwannian differentiation. Melanocytic schwannoma or malignant melanoma? AB - A 76 year old white woman presented with a four month history of dysphagia and weight loss. Clinical, radiological, and endoscopic examination revealed a pigmented mass in the lower third of the oesophagus. The preoperative diagnosis, including biopsy examination, was that of malignant melanoma. Following oesophageal resection, the mass was found to be a localised, relatively superficial tumour with light, electron microscopic, and immunohistochemical features common to both Schwann cells and melanocytes. The patient survived 46 months after surgery and died of a stroke, with no evidence of tumour recurrence. The tumour is presented as a case of melanocytic schwannoma, with unique features when compared with oesophageal melanotic schwannomas and malignant melanomas described in the literature. The differential diagnosis is discussed and an origin from a common precursor cell of neural crest origin is postulated. PMID- 11919223 TI - Thymidylate synthase levels: prognostic, predictive, or both? PMID- 11919224 TI - Pathologic evaluation of total mesorectal excision: big brother is watching. PMID- 11919225 TI - Referral to medical oncologists: are there barriers at the gate? PMID- 11919226 TI - Breast cancer consensus meetings: vive la difference? PMID- 11919227 TI - Thymidylate synthase expression in colorectal cancer: a prognostic and predictive marker of benefit from adjuvant fluorouracil-based chemotherapy. AB - PURPOSE: We studied the prognostic value of thymidylate synthase (TS) expression in primary colorectal cancer (CRC) and the role of TS expression as a predictor of chemotherapeutic benefit in patients treated with adjuvant chemotherapy. PATIENTS AND METHODS: TS expression was immunohistochemically assessed on tumor sections from 862 patients with CRC Dukes' stages B and C enrolled onto randomized trials evaluating fluorouracil (5-FU)-based adjuvant chemotherapy. RESULTS: TS expression was an independent prognostic factor for disease-free (P =.05) and overall survival (P =.05). In the subgroup treated with surgery alone, TS was an independent prognostic factor for disease-free (P <.001) and overall survival (P =.001), whereas this was not the case in the subgroup of adjuvantly treated patients. Patients whose tumors expressed high TS levels had a tendency to improved outcome after adjuvant therapy (not significant). The group whose tumors expressed the highest TS grade, grade 3 (34% of the patients), had a significantly longer disease-free survival if they were treated with adjuvant therapy compared with surgery alone (multivariate analyses, P =.02), whereas patients whose tumors expressed low TS levels (28% of the patients) had an impaired outcome after adjuvant therapy (multivariate analyses: disease-free survival, P =.01; overall survival, P =.01). CONCLUSION: TS expression predicts for survival independent of Dukes' stage in patients with CRC treated with surgery alone. The study indicates that patients with high TS levels may benefit from adjuvant 5-FU-based chemotherapy. However, patients with low TS levels seem to have a worse outcome when treated with adjuvant chemotherapy. PMID- 11919228 TI - Macroscopic evaluation of rectal cancer resection specimen: clinical significance of the pathologist in quality control. AB - PURPOSE: Quality assessment and assurance are important issues in modern health care. For the evaluation of surgical procedures, there are indirect parameters such as complication, recurrence, and survival rates. These parameters are of limited value for the individual surgeon, and there is an obvious need for direct parameters. We have evaluated criteria by which pathologists can judge the quality or completeness of the resection specimen in a randomized trial for rectal cancer. PATIENTS AND METHODS: The pathology reports of all patients entered onto a Dutch multicenter randomized trial were reviewed. All participating pathologists had been instructed by workshops and videos in order to obtain standardized pathology work-up. A three-tiered classification was applied to assess completeness of the total mesorectal excision (TME). Prognostic value of this classification was tested using log-rank analysis of Kaplan-Meier survival curves using the data of all patients who did not receive any adjuvant treatment. RESULTS: Included were 180 patients. In 24% (n = 43), the mesorectum was incomplete. Patients in this group had an increased risk for local and distant recurrence, 36.1% v. 20.3% recurrence in the group with a complete mesorectum (P =.02). Follow-up is too short to observe an effect on survival rates. CONCLUSION: A patient's prognosis is predicted by applying a classification of macroscopic completeness on a rectal resection specimen. We conclude that pathologists are able to judge the quality of TME for rectal cancer. With this direct interdisciplinary assessment instrument, we establish a new role of the pathologist in quality control. PMID- 11919229 TI - Investigation of the prognostic and predictive value of thymidylate synthase, p53, and Ki-67 in patients with locally advanced colon cancer. AB - PURPOSE: To evaluate the value of thymidylate synthase (TS), Ki-67, and p53 as prognostic markers in patients with Dukes' B2 and C colon carcinoma. METHODS: We conducted a retrospective analysis to evaluate the prognostic value of TS, Ki-67, and p53 in 465 patients with Dukes' B2 (220 patients) or Dukes' C (245 patients) colon carcinoma. Patients represent a nonrandom subset obtained from five randomized phase III trials and were treated with either surgery alone (151 patients) or surgery plus fluorouracil-based chemotherapy (314 patients). All three markers were assayed using immunohistochemical techniques. RESULTS: With a minimum follow-up of 5 years, our retrospective analysis failed to demonstrate a consistent and significant association between TS, Ki-67, or p53 and either disease-free survival or overall survival. Exploratory analyses did not reveal a convincing explanation for these results that are in conflict with the published literature. Notable interactions were observed. In particular, high Ki-67 levels were associated with increased (decreased) survival in patients with low (high) TS intensity. Patients whose tumors stained positively for p53 seemed to benefit substantially from the use of adjuvant chemotherapy compared with those who were not treated (P =.05). CONCLUSION: This retrospective investigation failed to demonstrate a significant association between TS, Ki-67, or p53 staining and clinical outcome. PMID- 11919231 TI - Randomized trial of postoperative adjuvant therapy in stage II and III rectal cancer to define the optimal sequence of chemotherapy and radiotherapy: a preliminary report. AB - PURPOSE: We conducted a prospective randomized trial to define the optimal sequence of chemotherapy and radiotherapy of postoperative adjuvant treatment in stage II and III rectal cancer. PATIENTS AND METHODS: Three hundred eight patients were enrolled onto the study. We randomly assigned 155 to arm I (early radiotherapy group) and 153 to arm II (late radiotherapy group). Treatment included eight cycles of chemotherapy at 4-week intervals and pelvic radiotherapy of 45 Gy in 25 fractions. Radiotherapy started on day 1 of the first chemotherapy cycle in arm I and on day 1 of the third chemotherapy cycle in arm II. The chemotherapy regimen consisted of fluorouracil 375 mg/m(2)/d and leucovorin 20 mg/m(2)/d. Chemotherapy was administered for 3 days per cycle in two cycles during the period of radiotherapy and for 5 days per cycle in the remaining six cycles. RESULTS: Twenty patients in arm I and 14 in arm II were not eligible. We included 274 patients in the analysis. With a median follow-up of 37 months for surviving patients, disease-free survival was significantly prolonged in arm I compared with arm II (81% v. 70% at 4 years; P =.043). Twenty-three recurrences occurred in arm I and 38 in arm II (P =.047). Overall survival was not significantly different between arms I and II (84% v. 82% at 4 years; P =.387). CONCLUSION: Early radiotherapy with concurrent chemotherapy after resection of stage II and III rectal cancer demonstrated a statistically significant advantage for disease-free survival compared with late radiotherapy with chemotherapy. PMID- 11919230 TI - Adjuvant therapy in rectal cancer: analysis of stage, sex, and local control- final report of intergroup 0114. AB - PURPOSE: The gastrointestinal Intergroup studied postoperative adjuvant chemotherapy and radiation therapy in patients with T3/4 and N+ rectal cancer after potentially curative surgery to try to improve chemotherapy and to determine the risk of systemic and local failure. PATIENTS AND METHODS: All patients had a potentially curative surgical resection and were treated with two cycles of chemotherapy followed by chemoradiation therapy and two additional cycles of chemotherapy. Chemotherapy regimens were bolus fluorouracil (5-FU), 5 FU and leucovorin, 5-FU and levamisole, and 5-FU, leucovorin, and levamisole. Pelvic irradiation was given to a dose of 45 Gy to the whole pelvis and a boost to 50.4 to 54 Gy. RESULTS: One thousand six hundred ninety-five patients were entered and fully assessable, with a median follow-up of 7.4 years. There was no difference in overall survival (OS) or disease-free survival (DFS) by drug regimen. DFS and OS decreased between years 5 and 7 (from 54% to 50% and 64% to 56%, respectively), although recurrence-free rates had only a small decrease. The local recurrence rate was 14% (9% in low-risk [T1 to N2+] and 18% in high-risk patients [T3N+, T4N]). Overall, 7-year survival rates were 70% and 45% for the low-risk and high-risk groups, respectively. Males had a poorer overall survival rate than females. CONCLUSION: There is no advantage to leucovorin- or levamisole containing regimens over bolus 5-FU alone in the adjuvant treatment of rectal cancer when combined with irradiation. Local and distant recurrence rates are still high, especially in T3N+ and T4 patients, even with full adjuvant chemoradiation therapy. PMID- 11919232 TI - Phase II study of capecitabine and oxaliplatin in first- and second-line treatment of advanced or metastatic colorectal cancer. AB - PURPOSE: To determine the efficacy and tolerability of combining oxaliplatin with capecitabine in the treatment of advanced nonpretreated and pretreated colorectal cancer. PATIENTS AND METHODS: Forty-three nonpretreated patients and 26 patients who had experienced one fluoropyrimidine-containing regimen for advanced colorectal cancer were treated with oxaliplatin 130 mg/m(2) on day 1 and capecitabine 1,250 mg/m(2) bid on days 1 to 14 every 3 weeks. Patients with good performance status (World Health Organization grade 0 to 1) were accrued onto two nonrandomized parallel arms of a phase II study. RESULTS: The objective response rate was 49% (95% confidence interval [CI], 33% to 65%) for nonpretreated and 15% (95% CI, 4% to 35%) for pretreated patients. The main toxicity of this combination was diarrhea, which occurred at grade 3 or 4 in 35% of the nonpretreated and 50% of the pretreated patients. Grade 3 or 4 sensory neuropathy, including laryngopharyngeal dysesthesia, occurred in 16% of patients on both cohorts. Capecitabine dose reductions were necessary in 26% of the nonpretreated and 45% of the pretreated patients in the second treatment cycle. The median overall survival was 17.1 months and 11.5 months, respectively. CONCLUSION: Combining capecitabine and oxaliplatin yields promising activity in advanced colorectal cancer. The main toxicity is diarrhea, which is manageable with appropriate dose reductions. On the basis of our toxicity experience, we recommend use of capecitabine in combination with oxaliplatin 130 mg/m(2) at an initial dose of 1,250 mg/m(2) bid in nonpretreated patients and at a dose of 1,000 mg/m(2) bid in pretreated patients. PMID- 11919233 TI - Acute oxaliplatin-induced peripheral nerve hyperexcitability. AB - PURPOSE: Oxaliplatin is a novel platinum compound with clinical activity in several malignancies. Neurotoxicity is dose-limiting and occurs in two distinct forms, an acute neurologic symptom complex that occurs within hours or days of therapy and a chronic, cumulative sensory neuropathy. PATIENTS AND METHODS: Patients were treated in a phase I study designed to establish the maximum tolerated dose of capecitabine given with oxaliplatin. Because of the unusual neurosensory toxicity of oxaliplatin, detailed neurologic examination, needle electromyography (EMG), and nerve conduction studies (NCS) were performed before and the day after oxaliplatin in a subset of 13 patients. Carbamazepine therapy was tried in 12 additional patients to determine whether the neurologic effects might be relieved. RESULTS: All patients experienced acute, reversible neurotoxicities with oxaliplatin. Symptoms included paresthesias, dysesthesias, cold hypersensitivity, jaw pain, eye pain, pain in the arm used for drug infusion, ptosis, leg cramps, and visual and voice changes. Serial EMG and NCS revealed striking signs of hyperexcitability in motor nerves after oxaliplatin. In patients who achieved therapeutic levels, carbamazepine did not alter the clinical or electromyographic abnormalities. CONCLUSION: The acute neurotoxicity seen with oxaliplatin is characterized by peripheral-nerve hyperexcitability, and the findings are similar to the clinical manifestations of neuromyotonia. Carbamezepine, which provides symptomatic relief in acquired neuromytonia, did not seem to be beneficial. Efforts to identify a successful neuroprotectant strategy would have a major impact on improving patient quality of life and the ability to deliver full doses of oxaliplatin. PMID- 11919234 TI - Tumor microvessel density as a predictor of recurrence after resection of hepatocellular carcinoma: a prospective study. AB - PURPOSE: This study prospectively evaluated the correlation of tumor microvessel density (MVD) with clinicopathologic features and postoperative recurrence in patients undergoing resection of hepatocellular carcinoma (HCC). PATIENTS AND METHODS: Tumor MVD was assessed in 100 patients with resection of HCC using a computer image analyzer after immunostaining for CD34 (MVD-CD34) and von Willebrand factor (MVD-vWF), respectively. Patients were prospectively followed for recurrence. RESULTS: Mean tumor MVD-CD34 (236/0.74 mm(2)) was higher than mean tumor MVD-vWF (87/0.74 mm(2)) (P <.001). By multiple regression analysis, tumor size was the only pathologic feature significantly related to tumor MVD CD34. The median MVD-CD34 was 316/0.74 mm(2) in HCCs < or = 5 cm (n = 46) and 146/0.74 mm(2) in HCCs more than 5 cm (n = 54) (P <.001). Among patients with HCCs < or = 5 cm, those with higher than median MVD-CD34 had worse disease-free survival (at 3 years, 13%) than those with a lower MVD-CD34 (at 3 year, 74%) (P =.002). Multivariate analysis showed that tumor MVD-CD34 was the only significant factor predictive of disease-free survival in patients with HCC < or = 5 cm. For HCCs more than 5 cm, MVD-CD34 did not have a significant prognostic influence. MVD-vWF did not have a significant prognostic influence on disease-free survival in either HCCs < or = 5 cm or more than 5 cm. CONCLUSION: This study shows that a high MVD-CD34 was predictive of early postresection recurrence in patients with HCCs < or = 5 cm and, therefore, may be a novel prognostic marker in this subset of patients. PMID- 11919235 TI - Impact of referral patterns on the use of chemotherapy for lung cancer. AB - PURPOSE: To determine the extent to which unexplained variation in the use of chemotherapy for advanced lung cancer is due to access to oncologists' services as opposed to treatment decisions made after seeing an oncologist. METHODS: We performed a retrospective cohort study of 12,015 patients over age 65 diagnosed with metastatic lung cancer between 1991 and 1996 while living in one of 11 regions monitored by a Survival, Epidemiology, and End Results (SEER) tumor registry. Assessment by an oncologist and subsequent treatment with chemotherapy were determined by examining linked Medicare claims. RESULTS: Of patients who did not receive chemotherapy, 36% were never assessed by a physician who provides chemotherapy. Patients living in certain areas, those diagnosed in more recent years, and those who received care in a teaching hospital were all more likely to see a cancer specialist. These factors were unrelated to subsequent treatment decisions, however. Conversely, age and comorbidity did not have a significant effect on whether a patient was seen by an oncologist, but they were associated with the likelihood of subsequently receiving chemotherapy. Black race, probably acting as a proxy for lower socioeconomic status, was associated with both a diminished likelihood of seeing a cancer specialist and subsequently receiving chemotherapy. CONCLUSION: Nonmedical factors are important determinants of whether a lung cancer patient is seen by a physician who provides chemotherapy. After seeing such a physician, treatment decisions seem to be mostly explained by appropriate medical factors. Racial and socioeconomic disparities still exist at both steps, however. As therapeutic options expand, referring physicians must ensure that biases and barriers to care do not deprive patients of the opportunity to consider all of their treatment options. PMID- 11919236 TI - Prognostic significance of the number of axillary lymph nodes removed in patients with node-negative breast cancer. AB - PURPOSE: The objective of the study was to evaluate the association between the number of lymph nodes removed at axillary dissection and recurrence and survival for patients with node-negative invasive breast cancer. PATIENTS AND METHODS: Subjects were 2,278 women with pathologically node-negative invasive breast cancer, diagnosed from 1989 to 1993 in British Columbia, Canada. Women aged > or = 90 years, with pure in-situ, bilateral invasive breast cancer or T4, N1, N2, or M1 stage, or who had axillary radiation were excluded. Two groups were defined for analysis: node-negative with no systemic therapy (n = 1,468) and node negative with systemic therapy (n = 810). Median follow-up was 7.5 years. Prognostic variables assessed were age at diagnosis, tumor size, tumor grade, invasion of lymphatics, veins, or nerves, estrogen receptor status, and number of nodes removed. RESULTS: For patients not receiving systemic therapy, regional relapse was significantly increased with smaller numbers of nodes removed (P =.03). There was a trend toward shorter overall survival with fewer nodes removed (P =.06). Node-negative patients who received systemic therapy did not have a higher regional relapse rate or shorter overall survival when fewer nodes were recovered. CONCLUSION: Recovery of a small number of negative lymph nodes at axillary dissection likely understages patients and leads to undertreatment, resulting in an increased regional relapse rate and poorer survival. The use of systemic therapy may overcome this effect. The number of nodes removed, in conjunction with other prognostic factors, may be useful in selecting node negative patients for systemic therapy. PMID- 11919237 TI - Phase II study of weekly docetaxel and trastuzumab for patients with HER-2 overexpressing metastatic breast cancer. AB - PURPOSE: To evaluate the safety and efficacy of weekly docetaxel plus trastuzumab in women with HER-2-overexpressing metastatic breast cancer. Efficacy was correlated with serum HER-2 extracellular domain (ECD) levels. PATIENTS AND METHODS: Thirty women with metastatic breast cancer were treated with weekly docetaxel and trastuzumab as first- or second-line therapy. Both docetaxel 35 mg/m(2)/wk and trastuzumab 2 mg/kg/wk were delivered in 4-week cycles consisting of three weekly treatments followed by 1 week of rest. A loading dose of trastuzumab 4 mg/kg was administered 1 day before the start of the first cycle. RESULTS: The median delivered dose-intensity of docetaxel was 24 mg/m(2)/wk (range, 18 to 27 mg/m(2)/wk). The intent-to-treat overall response rate (ORR) was 63% (95% confidence interval [CI], 44% to 80%). The ORR in patients whose tumors were HER-2-positive by fluorescence in situ hybridization was 67% (16 of 24 patients; 95% CI, 45% to 84%). In patients with elevated serum HER-2 ECD at baseline, the ORR was 76% (95% CI, 53% to 92%), compared with 33% (95% CI, 7% to 70%) in patients with low HER-2 ECD levels (P =.04). Variations in HER-2 ECD concentrations during treatment correlated with response to treatment. Median time to progression was 9 months. Acute toxicity, including myelosuppression, was mild. Fatigue, fluid retention, and excessive tearing became more common with repetitive dosing. CONCLUSION: Weekly docetaxel and trastuzumab is an active combination for treating patients with HER-2-overexpressing metastatic breast cancer. Serum HER-2 ECD testing may be a promising method for monitoring patients on trastuzumab-based therapy. PMID- 11919238 TI - Adjuvant therapy for breast cancer: practice patterns of community physicians. AB - PURPOSE: We evaluated the use of adjuvant therapy for breast cancer using the National Institutes of Health (NIH) Consensus Development Conference statements as guideposts for assessing how rapidly community physicians adopt recommended therapies. PATIENTS AND METHODS: Women with stage I through IIIA breast cancer diagnosed in 1987 through 1991 and in 1995 were randomly sampled from the population-based National Cancer Institute Surveillance, Epidemiology, and End Results program. A total of 8,106 women were included in the study with younger women, < or = 50 years, being oversampled. Their treating physicians were asked to verify whether chemotherapy, hormonal therapy, or both were given. RESULTS: After adjusting for clinical and nonclinical factors, the use of 1985 recommendations for adjuvant therapy in women with node-positive disease was already high at 80% in 1987 and increased slightly to 84% by 1995. Use of combined multidrug chemotherapy plus tamoxifen increased. In contrast, the use of 1990 recommendations for adjuvant therapy for node-negative disease was slightly less than 13% in 1987 and increased markedly to 57% by 1995. For women with node negative tumors > or = 1 cm in size diagnosed in 1995, 40% received tamoxifen, 16% combination chemotherapy, and 7% both, an increase from 10%, 5%, and 0.4%, respectively, in 1987. CONCLUSION: Community physicians began prescribing adjuvant chemotherapy and hormonal therapy in advance of publication of the NIH consensus statement in 1990. Adoption of recommended treatments for node-negative disease has been less complete compared with node-positive tumors, perhaps reflecting the more complex nature of the clinical trials data or the smaller anticipated benefit from adjuvant therapy for this disease subset. PMID- 11919239 TI - Interferon alfa therapy for malignant melanoma: a systematic review of randomized controlled trials. AB - PURPOSE: No standard systemic adjuvant therapy has been proven to increase overall survival in melanoma patients. The effect of interferon alfa (IFNalpha) as a single agent or in combination has been widely explored in clinical trials. The purpose of this study was to assess the benefit of IFNalpha therapy in malignant melanoma. METHODS: We performed a systematic review of randomized controlled trials comparing regimens with or without IFNalpha adjuvant therapy in melanoma patients. We assessed the effect of IFNalpha therapy on overall survival (OS), disease-free survival (DFS), melanoma recurrences, and toxicity. The quality of each trial was systematically evaluated. RESULTS: Nine randomized controlled trials (RCTs) of IFNalpha therapy in melanoma patients were identified. Eight were published and one was unpublished. Eight trials comprising 3,178 patients fulfilled our inclusion criteria and were analyzed. Quality assessment scores ranged from 22 to 71, with a mean score of 55.4 (95% confidence interval, 53.8 to 57.0). For OS, only one trial reported a statistically significant benefit for IFNalpha, but our analysis did not confirm it. Two trials reported statistically significant benefit in DFS for the patients treated with IFNalpha, but our analysis confirmed it in only one trial. There was a wide clinical heterogeneity between included trials, making meta-analysis inappropriate. CONCLUSION: In our review, results from included RCTs demonstrated no clear benefit of IFNalpha therapy on OS in melanoma patients. A large RCT is required to answer whether a full regimen of IFNalpha therapy is effective and to identify the subgroups of patients who might benefit from IFNalpha treatment. PMID- 11919240 TI - Tumor vascularity in the prognostic assessment of primary cutaneous melanoma. AB - PURPOSE: The vascular supply of the primary tumor is recognized to play an important role in the progression of a number of solid tumors. However, the role of tumor vascularity in the prognostic assessment of melanoma remains unclear. The purpose of this study was to determine the prognostic impact of patterns of vascularity on the outcome associated with cutaneous melanoma. PATIENTS AND METHODS: Tumor vascularity was documented prospectively using routine histopathologic analysis of 417 primary cutaneous melanomas from the University of California at San Francisco Melanoma Center database. Four patterns of tumor vascularity were recorded: absent, sparse, moderate, and prominent. RESULTS: Increasing tumor vascularity significantly increased the risk of relapse and death associated with melanoma, corresponding to reduced relapse-free and overall survival. By multivariate analysis, tumor vascularity was the most important determinant of overall survival, surpassing tumor thickness. Increasing tumor vascularity was associated with increased incidence of ulceration in the primary tumor. CONCLUSION: Tumor vascularity is an important prognostic factor in melanoma, rivaling tumor thickness. Increasing tumor vascularity is highly correlated with ulceration, possibly helping to explain the biologic basis of this known prognostic factor. PMID- 11919241 TI - Randomized trial of cisplatin and ifosfamide with or without bleomycin in squamous carcinoma of the cervix: a gynecologic oncology group study. AB - PURPOSE: Phase II trial reports have suggested that the addition of bleomycin to the combination of cisplatin and ifosfamide may improve response rates and possible survival in squamous carcinoma of the cervix. This study prospectively evaluates the combination of bleomycin to this regimen in women with histologically proven advanced recurrent or persistent squamous cell carcinoma of the cervix. PATIENTS AND METHODS: Eligible women were randomized to receive either cisplatin (50 mg/m(2)), ifosfamide (5 g/m(2) over 24 hours), and mesna (6 g/m(2) during ifosfamide infusion and the following 12 hours) (CI) versus bleomycin 30 units over 24 hours on day 1 followed by cisplatin (50 mg/m(2)), ifosfamide (5 g/m(2) over 24 hours), and mesna (6 g/m(2) during ifosfamide infusion and the following 12 hours) (CIB). Three hundred three women were enrolled onto this trial, of which 287 were assessable. RESULTS: There were no significant differences between CI and CIB with regard to response rates (32% v 31.2%, respectively), progression-free survival (PFS), or overall survival. PFS and survival were associated with initial performance status (PS). Patients with a PS of 0 experienced a lower rate of failure (P =.013) and a lower risk of death (P =.009) compared with patients with PS of 2. The most frequent grade 3/4 toxicities were leukopenia, neutropenia, anemia, thrombocytopenia, and nausea and vomiting. Neither regimen was associated with a significant increase in incidence of these toxicities. CONCLUSION: The CI regimen was virtually identical to CIB with regard to response rate, PFS, survival, and toxicity profile. Thus, the addition of bleomycin in the dose-schedule employed to cisplatin and ifosfamide did not improve outcome in patients with advanced cervical cancer. PMID- 11919242 TI - Low-risk persistent gestational trophoblastic disease: outcome after initial treatment with low-dose methotrexate and folinic acid from 1992 to 2000. AB - PURPOSE: We have simplified the treatment of gestational trophoblastic disease (GTD) in order to reduce the number of patients exposed to potentially carcinogenic chemotherapy. Patients who score 0 to 8 on the Charing Cross scoring system are classified as low-risk and receive methotrexate (MTX) and folinic acid (FA), whereas those who score higher than 8 are classified as high-risk and receive the etoposide, methotrexate, and dactinomycin (EMA)/cyclophosphamide and vincristine (CO) regimen. PATIENTS AND METHODS: Between 1992 and 2000, 485 women with GTD were commenced on MTX/FA at Charing Cross Hospital, London, United Kingdom. If patients developed MTX resistance or toxicity, treatment was altered according to the level of beta human chorionic gonadotropin (hCG). If serum hCG was < or = 100 IU/L, patients received dactinomycin; if hCG was greater than 100 IU/L, patients received EMA/CO. RESULTS: The median duration of follow-up was 4.7 years. Overall survival was 100% and the relapse rate was 3.3% (16 of 485 patients). hCG values normalized in 324 (66.8%) of 485 patients with MTX alone, whereas 161 (33.2%) of 485 patients required a change in treatment, 11 because of MTX toxicity and 150 because of MTX resistance. Sixty-seven patients changed to dactinomycin, of whom 58 achieved normal hCG values, and nine required third-line chemotherapy with EMA/CO. hCG values normalized in 93 (98.9%) of 94 patients who changed directly to EMA/CO from MTX. CONCLUSION: Single-agent dactinomycin has activity in patients with low-risk GTD who develop MTX resistance and whose hCG is low. Simplifying the stratification of GTD into two classes (low- and high risk) does not compromise overall outcome and may reduce the risk of second tumors. PMID- 11919243 TI - Excellent disease control and survival in patients with advanced nasopharyngeal cancer treated with chemoradiation. AB - PURPOSE: To determine the efficacy and safety of epirubicin, cisplatin, and infusional fluorouracil (5-FU) chemotherapy followed by radiation with concurrent cisplatin in patients with locally and/or regionally advanced nasopharyngeal cancer. PATIENTS AND METHODS: Thirty-five patients were treated with three cycles of induction chemotherapy with epirubicin 50 mg/m(2) and cisplatin 75 mg/m(2) combined with continuous-infusion 5-FU 200 mg/m(2) daily for 9 weeks, followed by concurrent chemoradiation of 60 Gy in 2-Gy fractions with cisplatin 20 mg/m(2) daily for 5 days in weeks 1 and 6. RESULTS: Median age was 43 years, 74% had World Health Organization type III histology, and 91% had stage IV disease (International Union Against Cancer, ed 4). All patients received three cycles of induction chemotherapy, and 97% completed chemoradiation. The estimated 4-year progression-free survival rate was 81% (95% CI, 59% to 93%), and the estimated 4 year overall survival rate was 90% (95% CI, 74% to 97%). Only two patients have had a locoregional relapse by the close-out date despite the use of only 60 Gy. Induction chemotherapy was well tolerated, with 11% grade 3 or 4 stomatitis, 26% grade 3 vomiting, and no episodes of febrile neutropenia. Acute toxicities of chemoradiation were as follows: 23% grade 3 or 4 vomiting, 6% febrile neutropenia, 31% grade 3 mucositis, and 23% grade 3 skin toxicity. The most prevalent grade 3 late effects were xerostomia and hearing loss. CONCLUSION: This regimen was well tolerated, can be delivered as planned, and has resulted in excellent locoregional disease control and survival in patients with locally advanced nasopharyngeal cancer. PMID- 11919244 TI - Risk of positive margins and biochemical recurrence in relation to nerve-sparing radical prostatectomy. AB - PURPOSE: To assess the effect of nerve-sparing (NS) radical retropubic prostatectomy (RRP) on surgical margins and biochemical recurrence. PATIENTS AND METHODS: Location and incidence of positive surgical margins, recurrence, and time to recurrence were assessed in a consecutive series of 734 men who underwent RRP for localized prostate cancer from 1992 through February 2000. NS procedures were used in 33% (n = 240) of 734 patients studied. RESULTS: Surgical margins were positive for 24% (n = 58) and 31% (n = 152) of NS and non-NS patients, respectively (P =.06). No significant difference between the groups was found in location of positive margins (P =.92). Prostate-specific antigen level greater than 10 ng/mL, extraprostatic extension, tumor volume more than 20%, capsular penetration, Gleason score > or = 7, positive margins, and seminal vesicle invasion were associated with significantly increased risk of recurrence. However, NS patients were not at increased risk of recurrence compared with non NS patients (hazard ratio, 0.96; 95% confidence interval, 0.53 to 1.72). The cumulative risk of recurrence within 3 and 5 years of surgery in NS patients was 9.7% and 14.4%, respectively, as compared with 17.1% and 21.1% for non-NS patients. CONCLUSION: In patients with localized prostate cancer, neither margin status nor biochemical-free survival within 5 years of surgery were altered by the nerve preservation technique. Given our experience, we recommend preservation of neurovascular bundles in these patients whenever the procedure is technically feasible. PMID- 11919246 TI - Extragonadal germ cell tumors of the mediastinum and retroperitoneum: results from an international analysis. AB - PURPOSE: To characterize the clinical and biologic features of extragonadal germ cell tumor (EGCT) and to determine the overall outcome with currently available treatment strategies. PATIENTS AND METHODS: Of an unselected population of 635 consecutive patients treated from 1975 through 1996 at 11 cancer centers, 341 patients (54%) had primary mediastinal EGCT, and 283 patients (45%) had retroperitoneal EGCT. Five hundred twenty-four patients (83%) had a nonseminomatous germ cell tumor (GCT), and 104 patients (16%) had a seminomatous histology. RESULTS: After platinum-based induction chemotherapy with or without secondary surgery, 141 patients (49%) with mediastinal nonseminomas (median follow-up, 19 months; range, 1 to 178 months) and 144 patients (63%) with retroperitoneal nonseminoma (median follow-up, 29 months; range, 1 to 203 months) are alive (P =.0006). In contrast, the overall survival rate for patients with a seminomatous EGCT is 88%, with no difference between patients with mediastinal or retroperitoneal tumor location (median follow-up, 49 months; range, 4 to 193 months; respective 70 months; range, 1 to 211 months). A significantly lower progression-free survival rate was found in seminoma patients treated with initial radiotherapy alone compared with chemotherapy. Nonseminomatous histology, presence of nonpulmonary visceral metastases, primary mediastinal GCT location, and elevated beta-human chorionic gonadotropin were independent prognostic factors for shorter survival. Hematologic malignancies (n = 17) occurred without exception in patients with primary mediastinal nonseminoma. Sixteen patients developed a metachronous testicular cancer despite the use of platinum-based chemotherapy. CONCLUSION: Whereas patients with pure seminomatous EGCT histology have a long-term chance of cure of almost 90% irrespective of the primary tumor site, 45% of patients with mediastinal nonseminomas are alive at 5 years. This outcome is clearly inferior compared with patients with nonseminomatous retroperitoneal primary tumors. PMID- 11919245 TI - Phase II study of paclitaxel plus gemcitabine in refractory germ cell tumors (E9897): a trial of the Eastern Cooperative Oncology Group. AB - PURPOSE: Despite great success in the treatment of disseminated germ cell tumors, 20% of patients are incurable and become candidates for investigational therapy. Paclitaxel and gemcitabine have shown activity as single agents in refractory germ cell tumors and can be combined with manageable toxicity. PATIENTS AND METHODS: Patients with germ cell tumors believed to be incurable with chemotherapy or surgery were treated with paclitaxel 110 mg/m(2) and gemcitabine 1,000 mg/m(2) intravenously on days 1, 8, and 15 of a 4-week cycle for a maximum of six cycles. Patients were evaluated for response and toxicity. RESULTS: Twenty eight of 30 enrolled patients were assessable. Toxicity was primarily hematologic but was manageable with only a single case of neutropenic fever. Six (21.4%) of 28 patients responded, including three complete responses. Two of the complete responders were continuously disease-free at 15+ and 25+ months. CONCLUSION: Paclitaxel plus gemcitabine is an active regimen in refractory germ cell tumors, with an acceptable toxicity profile. This regimen has the possibility for long term disease-free survival in this refractory patient population. PMID- 11919247 TI - Characterization of a novel prostate-specific antigen-activated peptide doxorubicin conjugate in patients with prostate cancer. AB - PURPOSE: To evaluate safety and pharmacokinetics (PK), and determine the recommended dose for efficacy studies, of L-377202, a novel peptide conjugate of doxorubicin (Dox) that releases the active metabolites leucine-doxorubicin (Leu Dox) and Dox on cleavage by membrane-bound prostate-specific antigen (PSA). PATIENTS AND METHODS: Nineteen patients with advanced hormone-refractory prostate cancer were treated intravenously with 71 cycles of L-377202 at escalating dose levels of 20 (n = 1), 40 (n = 3), 80 (n = 4), 160 (n = 3), 225 (n = 6), and 315 mg/m(2) (n = 2) once every 3 weeks. Toxicity, response, and PK of L-377202 were assessed. RESULTS: L-377202 was well tolerated. Dose-limiting grade 4 neutropenia was noted in two of two patients administered 315 mg/m(2) (both patients were able to resume therapy at 225 mg/m(2)). The recommended dose for efficacy studies was 225 mg/m(2), which induced grade 4 neutropenia in one of six patients. PK studies demonstrated that L-377202 was metabolized to Leu-Dox and Dox. PK were linear; after administration of single doses of 225 mg/m(2), the mean area under the concentration-time profiles of L-377202, Leu-Dox, and Dox were 6 micromol x L/h, 4 micromol x L/h, and 1 micromol x L/h, and peak concentrations were 14 micromol/L, 5 micromol/L, and 120 nmol/L, respectively. At 225 and 315 mg/m(2), five patients completed at least three cycles of therapy; two patients had a greater than 75% decrease in PSA, and one patient had a stabilized PSA. No response was noted at dose levels less than 225 mg/m(2). CONCLUSION: This is the first study of selective drug delivery in humans using a novel PSA-activated agent. L-377202 was cleaved to produce detectable levels of the active metabolites Leu-Dox and Dox. L-377202 was well tolerated and established a safe dose level for further study. PMID- 11919248 TI - Knowledge and experience regarding cancer, infertility, and sperm banking in younger male survivors. AB - PURPOSE: The goal of this study was to survey male patients aged 14 to 40 years at diagnosis and recently treated in two cancer centers to determine their knowledge, attitudes, and experiences regarding cancer-related infertility and sperm banking. PATIENTS AND METHODS: A postal survey about cancer-related infertility and sperm banking was offered to 904 men diagnosed with cancer within the previous 2 years. Eight percent opted out of the study. The others were sent the survey, with a cover letter stating elements of informed consent. RESULTS: Although the return rate was only 27%, yielding a sample of 201 men, responders did not differ significantly from nonresponders by institution, age, ethnicity, or cancer site. Overall, 51% of men wanted children in the future, including 77% of men who were childless at cancer diagnosis. Despite some anxieties about their own survival and risks to their children's health, men felt that the experience of cancer increased the value they placed on family closeness and would make them better parents. Only 60% of men recalled being informed about infertility as a side effect of cancer treatment, and just 51% had been offered sperm banking. Those who discussed infertility with their physicians had higher knowledge about cancer-related infertility and were significantly more likely to bank sperm. Only 24% of men banked sperm, including 37% of childless men. Lack of information was the most common reason for failing to bank sperm (25%). CONCLUSION: All men who are about to receive cancer treatment that could impair fertility should be counseled about such side effects and given adequate information to make an informed decision about banking sperm. PMID- 11919249 TI - Oncologists' attitudes and practices regarding banking sperm before cancer treatment. AB - PURPOSE: The goal of this study was to survey oncologists in three different practice settings to determine their knowledge, attitudes, and practices regarding referring patients to bank sperm before cancer treatment. METHODS: A postal survey about knowledge, attitudes, and practices regarding banking sperm before cancer treatment was sent to 718 oncology staff physicians and fellows at two cancer centers and at sites in a Community Clinical Oncology Program. RESULTS: The return rate was 24% and did not differ by institution, oncologic specialty, or sex. Fellows were significantly more likely to participate (37%) than staff physicians (20%). Ninety-one percent of respondents agreed that sperm banking should be offered to all men at risk of infertility as a result of cancer treatment, but 48% either never bring up the topic or mention it to less than a quarter of eligible men. Neither greater knowledge about sperm banking nor seeing large numbers of eligible men yearly increased the likelihood of discussing the option. Barriers cited included lack of time for the discussion, perceived high cost, and lack of convenient facilities. Oncologists reported they would be less likely to offer sperm banking to men who were homosexual, HIV-positive, had a poor prognosis, or had aggressive tumors. Oncologists overestimated the costs of sperm banking and the number of samples needed to make cryopreservation worthwhile. CONCLUSION: Sperm banking should be offered as an option to all men at risk of infertility because of their cancer treatment. Clearer practice standards could help oncologists increase their knowledge about sperm banking and avoid dependence on biased patient selection criteria. PMID- 11919250 TI - Aspergillus galactomannan detection in the diagnosis of invasive aspergillosis in cancer patients. AB - PURPOSE: To assess the Aspergillus galactomannan enzyme-linked immunosorbent assay (ELISA) in the diagnosis of invasive aspergillosis (IA) in adult and pediatric oncohematologic patients. PATIENTS AND METHODS: The study was conducted in four patient groups: those with fever of unknown origin (FUO) during neutropenia, suspected pulmonary infection (PI), or nonpulmonary aspergillosis (NPA) and those undergoing surveillance (S) after hematopoietic stem-cell transplantation (HSCT). IA was classified as definite, probable, or possible, according to European Organization for Research and Treatment of Cancer/Mycosis Study Group definitions. RESULTS: A total of 3,294 serum samples were collected during 797 episodes (FUO, 261; PI, 297; NPA, 28; and surveillance, 211), and 153 episodes of IA were diagnosed (31 definite, 67 probable, and 55 possible). Three episodes were first suspected from galactomannan ELISA; the remaining 150 cases were diagnosed from clinical or radiologic evidence. Sensitivity of the ELISA was 64.5%, 16.4%, and 25.5% in definite, probable, and possible episodes of IA, respectively, and was lower in patients positive for anti-Aspergillus antibodies than in antibody-negative patients. Most false-positive results occurred in children and in allogeneic HSCT (allo-HSCT) patients. Overall specificity of the ELISA was 94.8%. It was lower in children compared with adults (P <.0001) and in allo-HSCT patients compared with non-allo-HSCT adults (P =.0002). Lowering the ELISA cutoff value from 1.500 to 0.700 seemed more relevant for non-allo-HSCT adults (sensitivity, 73.1%, 44.3%, and 44.7% in definite, probable, and possible IA, respectively; specificity, 94%). CONCLUSION: Galactomannan ELISA seems less sensitive than previously described, and sensitivity can be further reduced by the presence of anti-Aspergillus antibodies. A new cutoff value for the ELISA of 0.700 is proposed for non-allo-HSCT adults. PMID- 11919251 TI - Psychiatric morbidity and impact on hospital length of stay among hematologic cancer patients receiving stem-cell transplantation. AB - PURPOSE: To determine the prevalence of psychiatric disorders during hospitalization for hematopoietic stem-cell transplantation (SCT) and to estimate their impact on hospital length of stay (LOS). PATIENTS AND METHODS: In a prospective inpatient study conducted from July 1994 to August 1997, 220 patients aged 16 to 65 years received SCT for hematologic cancer at a single institution. Patients received a psychiatric assessment at hospital admission and weekly during hospitalization until discharge or death, yielding a total of 1,062 psychiatric interviews performed. Psychiatric disorders were determined on the basis of the Diagnostic and Statistical Manual of Mental Disorders, Fourth Edition. Univariate and multivariate linear regression analyses were used to identify variables associated with LOS. RESULTS: Overall psychiatric disorder prevalence was 44.1%; an adjustment disorder was diagnosed in 22.7% of patients, a mood disorder in 14.1%, an anxiety disorder in 8.2%, and delirium in 7.3%. After adjusting for admission and in-hospital risk factors, diagnosis of any mood, anxiety, or adjustment disorder (P =.022), chronic myelogenous leukemia (P =.003), Karnofsky performance score less than 90 at hospital admission (P =.025), and higher regimen-related toxicity (P <.001) were associated with a longer LOS. Acute lymphoblastic leukemia (P =.009), non-Hodgkin's lymphoma (P =.04), use of peripheral-blood stem cells (P <.001), second year of study (P <.001), and third year of study (P <.001) were associated with a shorter LOS. CONCLUSION: Our data indicate high psychiatric morbidity and an association with longer LOS, underscoring the need for early recognition and effective treatment. PMID- 11919252 TI - Safe and cost effective use of alteplase for the clearance of occluded central venous access devices. AB - PURPOSE: To determine whether cryopreserved solutions of the thrombolytic agent alteplase could be used as a safe, effective, and economically reasonable alternative to urokinase in patients presenting with occluded central venous access devices (CVADs). MATERIALS AND METHODS: Alteplase has been reported as an efficacious alternative to urokinase for treatment of occluded CVADs. However, the practicality of using alteplase as the thrombolytic of choice for this indication remained conjectural. To make this approach economically feasible, alteplase was diluted to 1 mg/mL and 2.5-mL aliquots were stored at -20 degrees C until use. A need to confirm that the cryopreserving and thawing of the reconstituted solution did not compromise the safety and efficacy reported from prior trials was recognized. A quality assessment initiative was undertaken to concurrently monitor the safety and efficacy of this approach. Patients presenting with occluded CVADs received a sufficient volume of the thawed alteplase solution to fill the occluded catheter(s). Data, including efficacy, adverse reactions, dwell time, and catheter type, were collected over a 5-month period. RESULTS: One hundred twenty-one patients accounting for 168 attempted clearances were assessable for safety and efficacy. One hundred thirty-six (81%) of the 168 catheter clearance attempts resulted in successful catheter clearance (95% confidence interval, 74% to 86%). No adverse events were reported. CONCLUSION: Cryopreserved 1-mg/mL aliquots of alteplase are safe and effective in the clearance of occluded CVADs when stored at -20 degrees C for 30 days. The ability to cryopreserve alteplase aliquots makes it an economically reasonable alternative to urokinase in the setting of CVAD occlusion. PMID- 11919253 TI - Phase I and pharmacologic study of oral ZD9331, a novel nonpolyglutamated thymidylate synthase inhibitor, in adult patients with solid tumors. AB - PURPOSE: To assess the toxicity profile and dose-limiting toxicities (DLTs), to determine the maximum-tolerated dose, and to study the pharmacokinetics of ZD9331 when administered orally to patients with advanced solid tumors. PATIENTS AND METHODS: Patients were treated with oral ZD9331 given once daily (od) or twice daily (bid) for 5, 7, or 10 days; cycles were repeated every 21 days at doses ranging from 2.5 to 40 mg. For pharmacokinetic analysis, plasma sampling was performed during the first course and assayed using a validated liquid chromatographic-tandem mass spectrometry assay. Plasma levels of 2'-deoxyuridine were measured as a surrogate marker for TS inhibition. RESULTS: Forty-two patients received a total of 166 courses. The DLTs were myelosuppression and skin rash. Dose escalation of oral ZD9331 from 2.5 to 40 mg, as a single daily dose, resulted in a less than proportional increase in the plasma area under the concentration-time curve of ZD9331. The plasma drug exposure per cycle for the schedules 20 mg od for 5 days, 10 mg od for 10 days, and 10 mg bid for 5 days, all resulting in a total dose per cycle of 100 mg, were comparable. One partial response was noted in a patient with gastric cancer. CONCLUSION: DLTs in this phase I study of oral ZD9331 were myelosuppression and skin toxicity. The recommended dose for phase II studies of oral ZD9331 is 20 mg od for 5 consecutive days, every 3 weeks. PMID- 11919254 TI - DNA microarrays in clinical oncology. AB - Aberrant gene expression is critical for tumor initiation and progression. However, we lack a comprehensive understanding of all genes that are aberrantly expressed in human cancer. Recently, DNA microarrays have been used to obtain global views of human cancer gene expression and to identify genetic markers that might be important for diagnosis and therapy. We review clinical applications of these novel tools, discuss some important recent studies, identify promising avenues of research in this emerging field of study, and discuss the likely impact that expression profiling will have on clinical oncology. PMID- 11919255 TI - Unusual presentations of lymphoma: Case 1. Sea-blue histiocytes in non-Hodgkin's lymphoma. PMID- 11919256 TI - Unusual presentations of lymphoma: Case 2. Non-Hodgkin's lymphoma presenting as liver disease. PMID- 11919257 TI - Unusual presentations of lymphoma: Case 3. Splenic hematoma associated with non Hodgkin's lymphoma. PMID- 11919258 TI - Looking at the dying patient: the Ferdinand Hodler paintings of Valentine Gode Darel. PMID- 11919259 TI - Enhanced oral bioavailability of docetaxel by coadministration of cyclosporine: quantitation and role of P-glycoprotein. PMID- 11919260 TI - The art of oncology: when the tumor is not the target--the power of silence. PMID- 11919261 TI - High-dose samarium-153 ethylene diamine tetramethylene phosphonate: low toxicity of skeletal irradiation in patients with osteosarcoma and bone metastases. PMID- 11919262 TI - Equivalence between ovarian suppression and chemotherapy in the adjuvant treatment of endocrine-responsive breast cancer. PMID- 11919264 TI - Modulation of ecdysis in the moth Manduca sexta: the roles of the suboesophageal and thoracic ganglia. AB - The sequential behaviours shown by insects at ecdysis are due to the sequential release of various hormones, but the transition from one phase to the next can be fine-tuned by inhibitory influences. The ecdysis sequence in the moth Manduca sexta was initiated by injecting sensitive animals with the neuropeptide ecdysis triggering hormone (ETH). Exposure to ETH stimulates the release of eclosion hormone (EH) which, in turn, activates a set of neurons containing crustacean cardioactive peptide (CCAP) by elevating their levels of intracellular cyclic GMP. We characterized a set of non-CCAP containing neurons that also appear to be EH targets because of their response to cyclic GMP at ecdysis. The neurons did not display leucokinin-, diuretic-hormone- or FMRFamide-like immunoreactivity. They are probably the bursicon-containing cells described previously. After release of EH, there is a transient inhibition of the abdominal centers responsible for ecdysis. Transection experiments suggested that this suppression is via descending inhibitory units from the suboesophageal and thoracic ganglia. The duration of this inhibition appears to depend on the levels of cyclic GMP and can be extended by pharmacologically suppressing cyclic GMP breakdown. We further found that brief exposure to CO(2) caused premature ecdysis. Since the CO(2) treatment was effective only after EH release, it probably acts by suppressing descending inhibition. Studies on adult eclosion suggest that CO(2), given at the appropriate time, can uncouple the basic larval motor program from modulatory influences provided by the adult pterothoracic ganglion. CO(2) therefore appears to be a novel and non-invasive tool for studies of ecdysis behavior in insects. PMID- 11919263 TI - Meeting highlights: International consensus panel on the treatment of primary breast cancer. PMID- 11919265 TI - Expression of Manduca sexta V-ATPase genes mvB, mvG and mvd is regulated by ecdysteroids. AB - V-ATPases are complex proteins consisting of a peripheral, ATP-hydrolysing V(1) complex and a membrane-bound H(+)-translocating V(o) complex. The plasma membrane V-ATPase from the tobacco hornworm (Manduca sexta) midgut is made up of eight different V(1) and four different V(o) subunits. During starvation and moulting, V-ATPase activity decreases as a result of the dissociation of the V(1) complex from the V(o) complex. To determine whether subunit biosynthesis is reduced during periods of enzyme inactivity, we measured the transcript levels and transcriptional activities of V-ATPase genes. Northern blots revealed the downregulation of almost all V-ATPase transcripts during starvation. During moulting, transcript levels of the three V-ATPase genes examined, mvB, mvG and mvd, also decreased, and this decrease was negatively correlated with the titre of 20-hydroxyecdysone (20-HE) and positively correlated with the titre of juvenile hormone (JH). To test the biological significance of these correlations, we injected both hormones into feeding larvae and measured transcript levels several hours later. A short-term increase and a long-term decrease in levels of mRNA were observed after 20-HE injection, whereas JH injection had no significant effect. Immunohistochemical studies of the midgut epithelium revealed that 20-HE injection led to changes in goblet cell morphology and in the subcellular distribution of the V(1) complex comparable with the situation during the moult and during starvation. Reporter gene assays in Sf21 cells using mvB, mvG and mvd promoters to initiate transcription of firefly luciferase led, after incubation of the cells with 20-HE, to results comparable with those obtained in the injection experiments. These findings suggest that putative ecdysone-responsive elements are present in all three promoters. Taken together, our results suggest that the expression of V-ATPase genes is controlled in a coordinated manner by ecdysteroids. PMID- 11919267 TI - Reflections on colourful ommatidia of butterfly eyes. AB - The eye shine of butterflies from a large number of ommatidia was observed with a modified epi-illumination apparatus equipped with an objective lens of large numerical aperture. A few representative cases are presented: the satyrine Bicyclus anynana, the heliconian Heliconius melpomene, the small white Pieris rapae and the small copper Lycaena phlaeas. The colour of the eye shine is determined mainly by the reflectance spectrum of the tapetal mirror and the transmittance spectrum of the photoreceptor screening pigments, if present near the light-guiding rhabdom. Reflectance spectra measured from individual ommatidia show that tapetum and screening pigments are co-expressed in fixed combinations, thus determining different ommatidial classes. The classes are distributed in an irregular pattern that can be rapidly assessed with the novel epi-illumination apparatus. Many butterfly species appear to have red-reflecting ommatidia, which is interpreted to indicate the presence of red-sensitive photoreceptors. PMID- 11919266 TI - Determinants of intracellular pH in gas gland cells of the swimbladder of the European eel Anguilla anguilla. AB - Gas gland cells of the European eel (Anguilla anguilla) were cultured on collagen coated coverslips, and intracellular pH was measured using the pH-sensitive fluorescent probe 2',7'-bis-(2-carboxypropyl)-5-(6)-carboxyfluorescein (BCPCF). The contributions of various proton-translocating mechanisms to homeostasis of intracellular pH (pHi) were assessed by adding specific inhibitors of the various proton-translocating mechanisms at a constant extracellular pH (pHe) of 7.4 and after artificial acidification of the cells using the ammonium pulse technique. The greatest decrease in pHi was observed after addition of 5-(N-ethyl-N isobutyl)-amiloride (MIA), an inhibitor of Na(+)/H(+) exchange. Na(+)/H(+) exchange was active under steady-state conditions at an extracellular pH of 7.4, and activity increased after intracellular acidification. Incubation of gas gland cells with 4,4'-diisothiocyanostilbene-2,2'-disulphonic acid (DIDS), an inhibitor of anion exchange, also caused a decrease in pHi, but this decrease was not as pronounced as in the presence of MIA. Furthermore, at low pHi, the effect of DIDS was further reduced, suggesting that bicarbonate-exchanging mechanisms are involved in maintaining a steady-state pHi but that their importance is reduced at low pH. Bafilomycin A(1), a specific inhibitor of the V-ATPase, had no effect on steady-state pHi. However, recovery of intracellular pH after an artificial acid load was significantly impaired in the presence of bafilomycin. Our results suggest that Na(+)/H(+) exchange and anion exchange are important for the regulation of pHi at alkaline values of pHe. When pHi is low, a situation probably often encountered by gas gland cells during gas secretion, Na(+)/H(+) exchange continues to play an important role in acid secretion and a V-ATPase appears to contribute to proton secretion. PMID- 11919268 TI - The aerodynamic effects of wing rotation and a revised quasi-steady model of flapping flight. AB - We used a dynamically scaled model insect to measure the rotational forces produced by a flapping insect wing. A steadily translating wing was rotated at a range of constant angular velocities, and the resulting aerodynamic forces were measured using a sensor attached to the base of the wing. These instantaneous forces were compared with quasi-steady estimates based on translational force coefficients. Because translational and rotational velocities were constant, the wing inertia was negligible, and any difference between measured forces and estimates based on translational force coefficients could be attributed to the aerodynamic effects of wing rotation. By factoring out the geometry and kinematics of the wings from the rotational forces, we determined rotational force coefficients for a range of angular velocities and different axes of rotation. The measured coefficients were compared with a mathematical model developed for two-dimensional motions in inviscid fluids, which we adapted to the three-dimensional case using blade element theory. As predicted by theory, the rotational coefficient varied linearly with the position of the rotational axis for all angular velocities measured. The coefficient also, however, varied with angular velocity, in contrast to theoretical predictions. Using the measured rotational coefficients, we modified a standard quasi-steady model of insect flight to include rotational forces, translational forces and the added mass inertia. The revised model predicts the time course of force generation for several different patterns of flapping kinematics more accurately than a model based solely on translational force coefficients. By subtracting the improved quasi-steady estimates from the measured forces, we isolated the aerodynamic forces due to wake capture. PMID- 11919269 TI - Performance and adaptive value of tarsal morphology in rove beetles of the genus Stenus (Coleoptera, Staphylinidae). AB - To evaluate the adaptive value of the widening of the bilobed tarsi that has paralleled the tremendous radiation of the staphylinid genus Stenus, the performance of slender versus wide tarsi has been evaluated in two different contexts: (i) locomotion on the surface of water, and (ii) climbing on vertical (plant) surfaces. Contact angle measurements at the underside of the tarsi have revealed that, irrespective of tarsus width, all the investigated species are well supported by the surface of water while walking on it. The main selective demands driving the widening of the tarsi in several lineages have instead come from their firm attachment to smooth plant surfaces. This is suggested by measurements of the maximum vertical pulling forces exerted by intact and manipulated individuals on various rough and smooth surfaces. Species with widened tarsi associated with considerably more tenet setae attain significantly higher pulling forces, particularly on smooth surfaces. The tarsal setae are of greater importance on smooth surfaces, but the claws seem to be more important on rough substrata. On substrata that combine the attributes of rough and smooth surfaces, both claws and tenent setae add significantly to the pulling forces exerted, suggesting a functional synergism. The contribution of the present study to our understanding of insect tarsal attachment to surfaces with a variety of textures is discussed. PMID- 11919270 TI - Osmotic and volaemic effects on drinking rate in elasmobranch fish. AB - An increase in drinking rate of two species of marine elasmobranch fish, Scyliorhinus canicula and Triakis scyllia, acclimated to 80 % sea water was observed following the introduction of 100 % sea water to experimental tanks. The drinking response in both species was found to be maximal within 6 h, and a significant increase was sustained for up to 24 h in T. scyllia. Plasma osmolality was significantly increased within 6 h following introduction of 100 % sea water, and this increase was principally due to elevated plasma Na(+) and Cl( ) concentrations. Administration of 2 mol l(-1) mannitol, 75 % sucrose and vehicle (elasmobranch Ringer) did not induce a significant increase or decrease in the drinking rate of S. canicula. However, injection of 20 % NaCl was found to decrease drinking rate significantly in S. canicula 60 min after administration. Controlled haemorrhage of approximately 5.7 % of total blood volume in S. canicula induced a rapid 36-fold increase in drinking over basal levels. The present study demonstrates a physiological dipsogenesis in response to hypovolaemia in marine elasmobranch fish as part of their overall iso/hyperosmoregulatory strategy. PMID- 11919272 TI - Analysis of monoamines, adenosine and GABA in tissues of the land snail Helix lucorum and lizard Agama stellio stellio during hibernation. AB - The aim of the present study was to determine the levels of monoamines, GABA and adenosine in the brain, heart and haemolymph of the land snail Helix lucorum and in the brain, heart and blood of lizard Agama stellio stellio during long-term hibernation. We measured levels of the monoamines serotonin (5-HT) and its main metabolite 5-hydroxyindole-3-acetic acid (5-HIAA), dopamine (DA) and its metabolites dihydroxyphenylacetic acid (DOPAC) and homovanilic acid (HVA), norepinephrine (NE) and epinephrine (E). The most abundant amines detected in the brain and heart of active H. lucorum were 5-HT and DA. Of the metabolites examined only 5-HIAA was found in the brain. NE was found at very low levels but only in the brain, while E was not detected in the brain and heart. The levels of 5-HT and 5-HIAA increased in the brain and heart of H. lucorum within the first months of hibernation, showing a significant decrease thereafter. The levels of DA did not change during hibernation. The results indicated that 5-HT might be involved in preparing snails for entry into hibernation. GABA was only found in the brain of H. lucorum, and the levels were low; these levels remained during hibernation. Adenosine was present in brain and heart of H. lucorum, and during hibernation, the level of adenosine decreased significantly in the brain but remained steady in the heart. The monoamines 5-HT, DA and NE were present in the brain of active lizards A. stellio stellio, whereas E was found only at very low levels. Moreover, the metabolites 5-HIAA, DOPAC and HVA were detected in the brain of active lizards. The monoamines 5-HT, DA, NE and E were also detected in the heart and blood of active lizards. During hibernation the levels of these four monoamines were decreased significantly in the brain and heart of A. stellio stellio. In contrast, the levels of E increased in the heart and blood of hibernating lizards. Adenosine was detected in both heart and brain of active lizards, but hibernation caused a marked decrease in its levels at both tissues. GABA was found at higher levels than monoamines and adenosine in the brain of active lizards, and hibernation caused a significant increase in its levels, indicating an important role of GABA in inhibition of neuronal activity in hibernating lizards. PMID- 11919271 TI - Distribution and effects of PACAP, VIP, nitric oxide and GABA in the gut of the African clawed frog Xenopus laevis. AB - The distribution and possible effects on gastrointestinal motility of pituitary adenylate cyclase-activating polypeptide (PACAP), vasoactive intestinal polypeptide (VIP), nitric oxide and gamma-amino-butyric acid (GABA) were investigated in the African clawed frog (Xenopus laevis) using immunohistochemistry and in vitro strip preparations. PACAP- and VIP immunoreactive nerve fibres were common in the myenteric plexus as well as in the longitudinal and circular muscle layers all along the gastrointestinal tract. Double labelling demonstrated a close correlation between PACAP and VIP immunoreactivities, indicating that the two neurotransmitters are colocalised within the enteric nervous system. Occasionally, PACAP- and VIP-positive nerve cell bodies were seen in the myenteric or submucous plexa. In addition, VIP immunoreactivity coexisted with helospectin immunoreactivity. Nitric oxide synthase (NOS)-immunoreactive nerve cells were found in the myenteric plexus at an average density for the whole gastrointestinal tract of 4584 +/- 540 cells cm( 2). The NOS-immunoreactive nerve cells were usually multipolar with an average size of 11.3 +/- 3.7 x 23.2 +/- 6.6 microm. Some NOS-immunoreactive nerve fibres were VIP-immunoreactive but not all VIP-positive fibres showed NOS immunoreactivity. GABA immunoreactivity was found in nerve fibres and nerve cells in the myenteric plexus of all regions of the gut. Few GABA-immunoreactive nerve fibres were VIP-immunoreactive. PACAP 27, VIP, sodium nitroprusside (a nitric oxide donor; NaNP) and GABA caused similar responses on spontaneously contracting circular preparations of the cardiac stomach of X. laevis. The mean force developed was decreased, mainly by a reduction in resting tension, while the amplitude of contractions was not necessarily affected. The NOS inhibitor N(G) nitro-L-arginine methyl ester (L-NAME) increased the mean force developed, indicating a nitrergic tone in the preparations. In contrast, PACAP 27, VIP, NaNP, GABA and L-NAME had no significant effect on longitudinal strip preparations from the duodenum. These results indicate that PACAP, VIP, nitric oxide and GABA, which are known to be important inhibitory neurotransmitters in other vertebrates, are widely spread in the enteric nervous system of Xenopus laevis and may be involved in the inhibitory control of gastric motility. Although no effect of PACAP, VIP, nitric oxide or GABA on the longitudinal strips of the duodenum was seen in this study, this does not rule out the possibility that they might play an important role in controlling intestinal motility as well. PMID- 11919273 TI - Trade-offs between speed and endurance in the frog Xenopus laevis: a multi-level approach. AB - One of the most interesting trade-offs within the vertebrate locomotor system is that between speed and endurance capacity. However, few studies have demonstrated a conflict between whole-animal speed and endurance within a vertebrate species. We investigated the existence of trade-offs between speed and endurance capacity at both the whole-muscle and whole-animal levels in post-metamorphs of the frog Xenopus laevis. The burst-swimming performance of 55 frogs was assessed using a high-speed digital camera, and their endurance capacity was measured in a constant-velocity swimming flume. The work-loop technique was used to assess maximum power production of whole peroneus muscles at a cycle frequency of 6 Hz, while fatigue-resistance was determined by recording the decrease in force and net power production during a set of continuous cycles at 2 Hz. We found no significant correlations between measures of burst swimming performance and endurance capacity, suggesting that there is no trade-off between these two measures of whole-animal performance. In contrast, there was a significant negative correlation between peak instantaneous power output of the muscles at 6 Hz and the fatigue-resistance of force production at 2 Hz (other correlations between power and fatigue were negative but non-significant). Thus, our data support the suggestion that a physiological conflict between maximum power output and fatigue resistance exists at the level of vertebrate muscles. The apparent incongruence between whole-muscle and whole-animal performance warrants further detailed investigation and may be related to factors influencing both whole muscle and whole-animal performance measures. PMID- 11919274 TI - Changes in metabolic rate and N excretion in the marine invertebrate Sipunculus nudus under conditions of environmental hypercapnia: identifying effective acid base variables. AB - Increased CO(2) partial pressures (hypercapnia) as well as hypoxia are natural features of marine environments like the intertidal zone. Nevertheless little is known about the specific effects of CO(2) on metabolism, except for the well described effects on acid-base variables and regulation. Accordingly, the sediment-dwelling worm Sipunculus nudus was used as an experimental model to investigate the correlation of acid-base-induced metabolic depression and protein/amino acid catabolism, by determining the rates of oxygen consumption, ammonia excretion and O/N ratios in non-perfused preparations of body wall musculature at various levels of extra- and intracellular pH, P(CO(2)) and [HCO(3)(-)]. A decrease in extracellular pH from control level (7.9) to 6.7 caused a reduction in aerobic metabolic rate of both normocapnic and hypercapnic tissues by 40-45 %. O/N ratios of 4.0-4.5 under control conditions indicate that amino acid catabolism meets the largest fraction of aerobic energy demand. A significant 10-15 % drop in ammonia excretion, a simultaneous reduction of O/N ratios and a transient accumulation of intracellular bicarbonate during transition to extreme acidosis suggest a reduction in net amino acid catabolism and a shift in the selection of amino acids used, favouring monoamino dicarboxylic acids and their amines (asparagine, glutamine, aspartic and glutamic acids). A drop in intracellular pH was identified as mediating this effect. In conclusion, the present data provide evidence for a regulatory role of intracellular pH in the selection of amino acids used by catabolism. PMID- 11919275 TI - Regulation of the respiratory central pattern generator by chloride-dependent inhibition during development in the bullfrog (Rana catesbeiana). AB - Isolated brainstem preparations from larval (tadpole) and adult Rana catesbeiana were used to examine inhibitory mechanisms for developmental regulation of the respiratory central pattern generator (CPG). Preparations were superfused at 20 22 degrees C with Cl(-)-free artificial cerebrospinal fluid (aCSF) or with aCSF containing agonists/antagonists of gamma-aminobutyric acid (GABA) or glycine receptors. Respiratory motor output from the CPG, measured as neural activity from cranial nerve roots, was associated with fictive gill ventilation and lung ventilation in tadpoles and with fictive lung ventilation in adults. In tadpoles, fictive lung burst frequency was 0.8+/-0.2 min(-1) and did not change significantly with Cl(-)-free aCSF superfusion; however, lung burst amplitude increased by nearly 400 % (P<0.01). Fictive gill ventilation averaged 41.6+/-3.3 min(-1) and was reversibly abolished by Cl(-)-free aCSF. Superfusion with Cl(-) free aCSF abolished lung bursts in two of seven adult preparations, and overall lung burst frequency decreased from 3.1+/-0.7 to 0.4+/-0.03 min(-1) (P<0.01), but burst amplitude was unchanged. Low concentrations of GABA (0.5 mmol l(-1)) produced a significant increase in lung burst frequency followed by almost complete inhibition at 5.0 mmol l(-1), accompanied by the abolition of gill ventilation at 2.5-5.0 mmol l(-1). By contrast, fictive lung ventilation in adults was inhibited in a dose-dependent manner by glycine and GABA, and inhibition occurred at approximately 10-fold lower concentrations compared with tadpoles. The glycine receptor antagonist strychnine (2.5-25.0 micromol l(-1)) and the GABA(A) receptor antagonist bicuculline (1-10 micromol l(-1)) inhibited fictive gill ventilation and increased fictive lung ventilation in tadpoles. However, bicuculline and strychnine inhibited fictive lung ventilation in adults. These results suggest that lung ventilation in the tadpole brainstem may be driven by a pacemaker-like mechanism since Cl(-)-free aCSF failed to abolish lung ventilation. Lung ventilation in adults and gill ventilation in tadpoles, however, appear to be dependent upon conventional Cl(-)-mediated synaptic inhibition. Thus, there may be a developmental change in the fundamental process driving lung ventilation in amphibians. We hypothesize that maturation of the bullfrog respiratory CPG reflects developmental changes in glycinergic and/or GABAergic synaptic inhibitory mechanisms. PMID- 11919276 TI - The effects of continuous versus partial reinforcement schedules on associative learning, memory and extinction in Lymnaea stagnalis. AB - A continuous schedule of reinforcement (CR) in an operant conditioning procedure results in the acquisition of associative learning and the formation of long-term memory. A 50 % partial reinforcement (PR) schedule does not result in learning. The sequence of PR-CR training has different and significant effects on memory retention and resistance to extinction. A CR/PR schedule results in a longer lasting memory than a PR/CR schedule. Moreover, the memory produced by the CR/PR schedule is resistant to extinction training. In contrast, extinction occurs following the PR/CR schedule. PMID- 11919277 TI - Copper uptake across rainbow trout gills: mechanisms of apical entry. AB - Several components of branchial copper uptake were identified in juvenile freshwater rainbow trout (Oncorhynchus mykiss) using (64)Cu. On the basis of competitive interactions between sodium and copper uptake, inhibition of copper uptake by a proton pump inhibitor (bafilomycin A1, 2 micromol l(-1)) and a Na(+) channel blocker (phenamil, 100 micromol l(-1)), it appears that a proportion of the branchial copper uptake occurs via an apical Na(+) channel. This sodium sensitive copper uptake demonstrates saturation kinetics, with a K(m) of 7.1 nmol l(-1) and a J(max) of 21.2 pmol g(-1) h(-1), and is characterized by an IC(50) of 104 micromol l(-1) sodium. On the basis of residual copper uptake in the presence of high sodium concentrations (20 mmol l(-1)) and differential inhibition of sodium and copper uptake by phenamil (100 micromol l(-1)), a sodium-insensitive component of copper uptake is also present in trout gills. It demonstrates saturation kinetics with a comparably low K(m) (9.6 nmol l(-1)) but a lower maximum transport capacity (J(max)=3.5 pmol g(-1) h(-1)) than the sodium insensitive system. Sodium uptake exhibits saturation kinetics with a K(m) of 69 micromol l(-1). Copper reduced branchial sodium transport affinity but increased the maximal sodium transport capacity. PMID- 11919278 TI - Phosphoglucose isomerases of hagfish, zebrafish, gray mullet, toad, and snake, with reference to the evolution of the genes in vertebrates. AB - Phosphoglucose isomerase (PGI) is a protein with multiple functions. To infer its structure changes and evolution in vertebrates, we cloned cDNAs encoding PGI genes from hagfish (Paramyxine yangi), gray mullet (Mugil cephalus), zebrafish (Danio rerio), toad (Bufo melanosticus), and snake (Boiga kraepelini). Only one PGI gene was cloned in each of hagfish, toad, and snake, but two PGI genes were found in zebrafish and gray mullet, respectively. The PGI of hagfish encodes 554 amino acids, in contrast to the PGIs of bonyfishes, toad, and snake which encode 553 amino acids and the PGIs of mammals which encode 558 amino acids. Among 558 aligned amino acid sites, there are 314 sites (56.27%) totally conserved. To see if diversifying selection acts on PGI amino acids of vertebrates, we calculated the pairwise ratio of nonsynonymous versus synonymous substitution per site (Ka/Ks) and the ratio of radical amino acid changes versus conservative amino acid changes per sites (dR/dC) between PGI sequences. The average pairwise ratio between nonsynonymous substitutions per nucleotide (Ka) and synonymous substitutions per nucleotide (Ks) among vertebrate PGI sequences equals 0.047 +/- 0.019. The average pairwise ratio between radical amino acid changes and conservative amino acid changes (dR/dC) among the vertebrate PGIs equal 0.938 +/- 0.158 for charge changes, 0.558 +/- 0.085 for polarity changes, and 0.465 +/- 0.0714 when both polarity and volume are considered. There is no amino acid within the vertebrate PGIs under diversifying selection as analyzed by the method of Yang et al. (2000b). The results suggest that the present vertebrate PGIs are at evolutionary stasis and are being subjected to intense purifying selection. The purifying selection is to maintain polarity and volume of the protein but not the charge groups of amino acids. Phylogenetic analysis reveals that vertebrate PGIs can be classified into three major groups: the mammalian, amphibian reptilian, and teleostean PGIs. The gene tree suggests that the gene duplication event of PGI in bonyfishes occurred before diversification of Acanthopterygii but after the split of bonyfishes and tetrapods. The evolution of multiple functions of PGI is discussed. PMID- 11919280 TI - The effects of nucleotide substitution model assumptions on estimates of nonparametric bootstrap support. AB - The use of parameter-rich substitution models in molecular phylogenetics has been criticized on the basis that these models can cause a reduction both in accuracy and in the ability to discriminate among competing topologies. We have explored the relationship between nucleotide substitution model complexity and nonparametric bootstrap support under maximum likelihood (ML) for six data sets for which the true relationships are known with a high degree of certainty. We also performed equally weighted maximum parsimony analyses in order to assess the effects of ignoring branch length information during tree selection. We observed that maximum parsimony gave the lowest mean estimate of bootstrap support for the correct set of nodes relative to the ML models for every data set except one. For several data sets, we established that the exact distribution used to model among site rate variation was critical for a successful phylogenetic analysis. Site specific rate models were shown to perform very poorly relative to gamma and invariable sites models for several of the data sets most likely because of the gross underestimation of branch lengths. The invariable sites model also performed poorly for several data sets where this model had a poor fit to the data, suggesting that addition of the gamma distribution can be critical. Estimates of bootstrap support for the correct nodes often increased under gamma and invariable sites models relative to equal rates models. Our observations are contrary to the prediction that such models cause reduced confidence in phylogenetic hypotheses. Our results raise several issues regarding the process of model selection, and we briefly discuss model selection uncertainty and the role of sensitivity analyses in molecular phylogenetics. PMID- 11919279 TI - Evolutionary implications of three novel members of the human sarcomeric myosin heavy chain gene family. AB - Sarcomeric myosin heavy chain (MyHC) is the major contractile protein of striated muscle. Six tandemly linked skeletal MyHC genes on chromosome 17 and two cardiac MyHC genes on chromosome 14 have been previously described in the human genome. We report the identification of three novel human sarcomeric MyHC genes on chromosomes 3, 7, and 20, which are notable for their atypical size and intron exon structure. Two of the encoded proteins are structurally most like the slow beta MyHC, whereas the third one is closest to the adult fast IIb isoform. Data from pairwise comparisons of aligned coding sequences imply the existence of ancestral genomes with four sarcomeric genes before the emergence of a dedicated smooth muscle MyHC gene. To further address the evolutionary relationships of the distinct sarcomeric and nonsarcomeric rod sequences, we have identified and further annotated human genomic DNA sequences corresponding to 14 class-II MyHCs. An extensive analysis provides a timeline for intron gain and loss, gene contraction and expansion, and gene conversion among genes encoding class-II myosins. One of the novel human genes is found to have introns at positions shared only with the molluscan catchin/MyHC gene, providing evidence for the structure of a pre-Cambrian ancestral gene. PMID- 11919281 TI - Electron transfer in ferredoxin: are tunneling pathways evolutionarily conserved? AB - A theoretical study of electron transfer (ET) pathways in a recently crystallized Clostridium acidurici ferredoxin is reported. The electronic structure of the protein complex is treated at the semiempirical extended Huckel level, and the tunneling pathways are calculated with the rigorous quantum mechanical method of tunneling currents. The model predicts two pathways between the two [4Fe-4S] cubanes: a strong one running directly from Cys(14) to Cys(43) and a weaker one from Cys(14) via Ile(23) to Cys(18), whereas other amino acids do not play a significant role in the electron tunneling. The cysteine ligands conduct almost all of the current when Ile(23) is mutated to valine in silico, so that there is no appreciable change in the ET rate. The calculated value of the transfer matrix element is consistent with the experimentally determined rate of transfer. Results of the sequence analysis performed on this ferredoxin reveal that Ile(23) is a highly variable amino acid compared with the cubane-ligating cysteine amino acids, even though Ile(23) lies directly between the donor and acceptor complexes. We further argue that the homologous proteins with a [3Fe-4S] cofactor, which does not have one of the four cysteine ligands, use the same tunneling pathways as those in this ferredoxin, on the basis of the high homology as well as the absolute conservation of Cys(14) and Cys(43) which serve as the main tunneling conduit. Our results explain why mutation of amino acids around and between the donor and acceptor cubane clusters, including that of Ile(23), does not appreciably affect the rate of transfer and add support to the proposal that there exist evolutionarily conserved electron tunneling pathways in biological ET reactions. PMID- 11919282 TI - Cytoglobin: a novel globin type ubiquitously expressed in vertebrate tissues. AB - Vertebrates possess multiple respiratory globins that differ in terms of structure, function, and tissue distribution. Three types of globins have been described so far: hemoglobin facilitates the transport of oxygen in the blood, myoglobin serves oxygen transport and storage in the muscle, and neuroglobin has a yet unidentified function in nerve cells. Here we report the identification of a fourth and novel type of globin in mouse, man, and zebrafish. It is expressed in apparently all types of human tissue and therefore has been called cytoglobin (CYGB). Mouse and human CYGBs comprise 190 amino acids; the zebrafish CYGB, 174 amino acids. The human CYGB gene is located on chromosome 17q25. The mammalian genes display a unique exon-intron pattern with an additional exon resulting in a C-terminal extension of the protein, which is absent in the fish CYGB. Phylogenetic analyses suggest that the CYGBs had a common ancestor with vertebrate myoglobins. This indicates that the vertebrate myoglobins are in fact a specialized intracellular globin that evolved in adaptation to the special needs of muscle cells. PMID- 11919283 TI - The chaperonin genes of jakobid and jakobid-like flagellates: implications for eukaryotic evolution. AB - The jakobids are free-living mitochondriate protists that share ultrastructural features with certain amitochondriate groups and possess the most bacterial-like mitochondrial genomes described thus far. Jakobids belong to a diverse group of mitochondriate and amitochondriate eukaryotes, the excavate taxa. The relationships among the various excavate taxa and their relationships to other putative deep-branching protist groups are largely unknown. With the hope of clarifying these issues, we have isolated the cytosolic chaperonin CCTalpha gene from the jakobid Reclinomonas americana (strains 50394 and 50283), the jakobid like malawimonad Malawimonas jakobiformis, two heteroloboseans (Acrasis rosea and Naegleria gruberi), a euglenozoan (Trypanosoma brucei), and a parabasalid (Monocercomonas sp.). We also amplified the CCTdelta gene from M. jakobiformis. The Reclinomonas and Malawimonas sequences presented here are among the first nuclear protein-coding genes to be described from these organisms. Unlike other putative early diverging protist lineages, a high density of spliceosomal introns was found in the jakobid and malawimonad CCTs-similar to that observed in vertebrate protein-coding genes. An analysis of intron positions in CCT genes from protists, plants, animals, and fungi suggests that many of the intron-sparse or intron-lacking protist lineages may not be primitively so but have lost spliceosomal introns during their evolutionary history. In phylogenetic trees constructed from CCTalpha protein sequences, R. americana (but not M. jakobiformis) shows a weak but consistent affinity for the Heterolobosea and Euglenozoa. PMID- 11919284 TI - When does the incongruence length difference test fail? AB - This paper examines the efficiency of the incongruence length difference test (ILD) proposed by Farris et al. (1994) for assessing the incongruence between sets of characters. DNA sequences were simulated under various evolutionary conditions: (1) following symmetric or asymmetric trees, (2) with various mutation rates, (3) with constant or variable evolutionary rates along the branches, and (4) with different among-site substitution rates. We first compared two sets of sequences generated along the same tree and under the same evolutionary conditions. The probability of a Type-I error (wrongly rejecting the true hypothesis of congruence) was substantially below the standard 5% level of significance given by the ILD test; this finding indicates that the choice of the 5% level is rather conservative in this case. We then compared two data sets, still generated along the same tree, but under different evolutionary conditions (constant vs. variable evolutionary rate, homogeneity vs. heterogeneity rate of substitution). Under these conditions, the probability of rejecting the true hypothesis of congruence was greater than the 5% given by the ILD test and increased with the number of sites and the degree to which the tree was asymmetric. Finally, the comparison of the two data sets, simulated under contrasting tree structures (symmetric vs. asymmetric) but under the same evolutionary conditions, led us to reject the hypothesis of congruence, albeit weakly, particularly when the number of informative sites was low and among-site substitution rate heterogeneous. We conclude that the ILD test has only limited power to detect incongruence caused by differences in the evolutionary conditions or in the tree topology, except when numerous characters are present and the substitution rate is homogeneous from site to site. PMID- 11919285 TI - Diversifying selection of the tumor-growth promoter angiogenin in primate evolution. AB - Diversifying selection drives the rapid differentiation of gene sequences and is one of the main forces behind adaptive evolution. Most genes known to be shaped by diversifying selection are those involved in host-pathogen or male-female interactions characterized as molecular "arms races." Here we report the unexpected detection of diversifying selection in the evolution of a tumor-growth promoter, angiogenin (ANG). A comparison among 11 primate species demonstrates that ANG has a significantly higher rate of nucleotide substitution at nonsynonymous sites than at synonymous sites, a hallmark of positive selection acting at the molecular level. Furthermore, we observed significant charge diversity at the molecular surface, suggesting the presence of selective pressures in the microenvironment of ANG, including its binding molecules. A population survey of ANG in chimpanzees, however, reveals no polymorphism, which may have resulted from a recent selective sweep of a charge-altering substitution in chimpanzee evolution. Functional assays of recombinant ANGs from the human and owl monkey indicate that primate ANGs retain angiogenic activity despite rapid evolution. Our study, together with findings of similar selection in the primate breast cancer suppressor gene, BRCA1, reveals an intriguing phenomenon of unusual selective pressures on, and adaptive evolution of, cancer-related genes in primate evolution. PMID- 11919286 TI - Evidence for multiple genetic forms with similar eyeless phenotypes in the blind cavefish, Astyanax mexicanus. AB - A diverse group of animals has adapted to caves and lost their eyes and pigmentation, but little is known about how these animals and their striking phenotypes have evolved. The teleost Astyanax mexicanus consists of an eyed epigean form (surface fish) and at least 29 different populations of eyeless hypogean forms (cavefish). Current alternative hypotheses suggest that adaptation to cave environments may have occurred either once or multiple times during the evolutionary history of this species. If the latter is true, the unique phenotypes of different cave-dwelling populations may result from convergence of form, and different genetic changes and developmental processes may have similar morphological consequences. Here we report an analysis of variation in the mitochondrial NADH dehydrogenase 2 (ND2) gene among different surface fish and cavefish populations. The results identify a minimum of two genetically distinctive cavefish lineages with similar eyeless phenotypes. The distinction between these divergent forms is supported by differences in the number of rib bearing thoracic vertebrae in their axial skeletons. The geographic distribution of ND2 haplotypes is consistent with roles for multiple founder events and introgressive hybridization in the evolution of cave-related phenotypes. The existence of multiple genetic lineages makes A. mexicanus an excellent model to study convergence and the genes and developmental pathways involved in the evolution of the eye and pigment degeneration. PMID- 11919287 TI - A new subfamily of major intrinsic proteins in plants. AB - The major intrinsic proteins (MIPs) form a large protein family of ancient origin and are found in bacteria, fungi, animals, and plants. MIPs act as channels in membranes to facilitate passive transport across the membrane. Some MIPs allow small polar molecules like glycerol or urea to pass through the membrane. However, the majority of MIPs are thought to be aquaporins (AQPs), i.e., they are specific for water transport. Plant MIPs can be subdivided into the plasma membrane intrinsic protein, tonoplast intrinsic protein, and NOD26-like intrinsic protein subfamilies. By database mining and phylogenetic analyses, we have identified a new subfamily in plants, the Small basic Intrinsic Proteins (SIPs). Comparisons of sequences from the new subfamily with conserved amino acid residues in other MIPs reveal characteristic features of SIPs. Possible functional consequences of these features are discussed in relation to the recently solved structures of AQP1 and GlpF. We suggest that substitutions at conserved and structurally important positions imply a different substrate specificity for the new subfamily. PMID- 11919288 TI - The complex history of a gene proposed to participate in a sexual isolation mechanism in house mice. AB - Previous behavioral experiments showed that mouse salivary androgen-binding protein (ABP) was involved in interindividual recognition and might play a role in sexual isolation between house mouse (Mus musculus) subspecies. The pattern of evolution of Abpa, the gene for the alpha subunit of ABP, was found to be consistent with this hypothesis. Abpa apparently diverged rapidly between species and subspecies with a large excess of nonsynonymous substitutions, a lack of exon polymorphism within each of the three subspecies, and a lack of intron polymorphism in the one subspecies studied (M. musculus domesticus). Here we characterized the intron and exon sequence variations of this gene in house mouse populations from central Eurasia, a region yet unsampled and thought to be close to the cradle of the radiation of the subspecies. We also determined the intron and exon sequences in seven other species of the genus Mus. We confirmed the general pattern of rapid evolution by essentially nonsynonymous substitutions, both inter- and intraspecifically, supporting the idea that Darwinian selection has driven the evolution of this gene. We also observed a uniform intron sequence in five samples of M. musculus musculus, suggesting that a selective sweep might have occurred for that allele. In contrast to previous results, however, we found extensive intron and exon polymorphism in some house mouse populations from central Eurasia. We also found evidence for secondary admixture of the subspecies specific alleles in regions of transition between the subspecies in central Eurasia. Furthermore, an abnormal intron phylogeny suggested that interspecific exchanges had occurred between the house mouse subspecies and three other Palearctic species. These observations appear to be at variance with the simple hypothesis that Abpa is involved in reproductive isolation. Although we do not rule out a role in recognition, the situation appears to be more complex than previously thought. Thus the selective mechanism behind the evolution of Abpa remains to be resolved, and we suggest that it may have changed during the recent colonization history of the house mouse. PMID- 11919290 TI - Evolution of dinoflagellate unigenic minicircles and the partially concerted divergence of their putative replicon origins. AB - Dinoflagellate chloroplast genes are unique in that each gene is on a separate minicircular chromosome. To understand the origin and evolution of this exceptional genomic organization we completely sequenced chloroplast psbA and 23S rRNA gene minicircles from four dinoflagellates: three closely related Heterocapsa species (H. pygmaea, H. rotundata, and H. niei) and the very distantly related Amphidinium carterae. We also completely sequenced a Protoceratium reticulatum minicircle with a 23S rRNA gene of novel structure. Comparison of these minicircles with those previously sequenced from H. triquetra and A. operculatum shows that in addition to the single gene all have noncoding regions of approximately a kilobase, which are likely to include a replication origin, promoter, and perhaps segregation sequences. The noncoding regions always have a high potential for folding into hairpins and loops. In all six dinoflagellate strains for which multiple minicircles are fully sequenced, parts of the noncoding regions, designated cores, are almost identical between the psbA and 23S rRNA minicircles, but the remainder is very different. There are two, three, or four cores per circle, sometimes highly related in sequence, but no sequence identity is detectable between cores of different species, even within one genus. This contrast between very high core conservation within a species, but none among species, indicates that cores are diverging relatively rapidly in a concerted manner. This is the first well-established case of concerted evolution of noncoding regions on numerous separate chromosomes. It differs from concerted evolution among tandemly repeated spacers between rRNA genes, and that of inverted repeats in plant chloroplast genomes, in involving only the noncoding DNA cores. We present two models for the origin of chloroplast gene minicircles in dinoflagellates from a typical ancestral multigenic chloroplast genome. Both involve substantial genomic reduction and gene transfer to the nucleus. One assumes differential gene deletion within a multicopy population of the resulting oligogenic circles. The other postulates active transposition of putative replicon origins and formation of minicircles by homologous recombination between them. PMID- 11919291 TI - Origin and evolution of influenza virus hemagglutinin genes. AB - Influenza A, B, and C viruses are the etiological agents of influenza. Hemagglutinin (HA) is the major envelope glycoprotein of influenza A and B viruses, and hemagglutinin-esterase (HE) in influenza C viruses is a protein homologous to HA. Because influenza A virus pandemics in humans appear to occur when new subtypes of HA genes are introduced from aquatic birds that are known to be the natural reservoir of the viruses, an understanding of the origin and evolution of HA genes is of particular importance. We therefore conducted a phylogenetic analysis of HA and HE genes and showed that the influenza A and B virus HA genes diverged much earlier than the divergence between different subtypes of influenza A virus HA genes. The rate of amino acid substitution for A virus HAs from duck, a natural reservoir, was estimated to be 3.19 x 10(-4) per site per year, which was slower than that for human and swine A virus HAs but similar to that for influenza B and C virus HAs (HEs). Using this substitution rate from the duck, we estimated that the divergences between different subtypes of A virus HA genes occurred from several thousand to several hundred years ago. In particular, the earliest divergence time was estimated to be about 2,000 years ago. Also, the A virus HA gene diverged from the B virus HA gene about 4,000 years ago and from the C virus HE gene about 8,000 years ago. These time estimates are much earlier than the previous ones. PMID- 11919292 TI - Evolution of the Fot1 transposons in the genus Fusarium: discontinuous distribution and epigenetic inactivation. AB - To understand the evolution of Fot1, a member of the pogo family widely dispersed in ascomycetes, we have performed a phylogenetic survey across the genus Fusarium divided into six sections. The taxonomic distribution of Fot1 is not homogeneous but patchy; it is prevalent in the Fusarium oxysporum complex, absent in closely related sections, and found in five species from the most distant section Martiella. Multiple copies of Fot1 were sequenced from each strain in which the element occurs. In three species, the Fot1 nucleotide sequence is 98% identical to that from F. oxysporum (Fox), whereas nucleotide divergence for host genes is markedly higher: 11% for partial nuclear 28S rDNA and up to 30% for the gene encoding nitrate reductase (nia). In two species, sequence divergence of Fot1 related elements relative to Fox ranged from 7% to 23% (16% average). Most of the sequence differences (82%) were C-to-T and G-to-A transitions. These mutations are distributed throughout the Fot1 sequences, although they tend to be concentrated in the middle portion of the elements. Analysis of the local sequence context of transitions revealed a hierarchy of site preferences. These characteristics are typical of the repeat-induced point mutation process, first discovered in Neurospora crassa. The spotty distribution of Fot1 elements among species together with the high degree of similarity between Fot1 sequences present in distant species strongly suggests a case of horizontal transfer. PMID- 11919293 TI - Do introns favor or avoid regions of amino acid conservation? AB - Are intron positions correlated with regions of high amino acid conservation? For a set of ancient conserved proteins, with intronless prokaryotic but intron containing eukaryotic homologs, multiple sequence alignments identified residues invariant throughout evolution. Intron positions between codons show no preferences. However, introns lying after the first base of a codon prefer conserved regions, markedly in glycines. Because glycines are in excess in conserved regions, this behavior could reflect phase-one introns entering glycine residues randomly in the ancestral sequences. Examination of intron positions within codons of evolutionarily invariable amino acids showed that roughly 50% of these introns are bordered by guanines at both 5'- and 3'-ends, 25% have a G only before the intron, and 5% have a G only after the intron, whereas about 20% are bordered by nonguanine bases. PMID- 11919289 TI - Inferring the history of speciation from multilocus DNA sequence data: the case of Drosophila pseudoobscura and close relatives. AB - The divergence of Drosophila pseudoobscura from its close relatives, D. persimilis and D. pseudoobscura bogotana, was examined using the pattern of DNA sequence variation in a common set of 50 inbred lines at 11 loci from diverse locations in the genome. Drosophila pseudoobscura and D. persimilis show a marked excess of low-frequency variation across loci, consistent with a model of recent population expansion in both species. The different loci vary considerably, both in polymorphism levels and in the levels of polymorphisms that are shared by different species pairs. A major question we address is whether these patterns of shared variation are best explained by gene flow or by persistence since common ancestry. A new test of gene flow, based on patterns of linkage disequilibrium, is developed. The results from these, and other tests, support a model in which D. pseudoobscura and D. persimilis have exchanged genes at some loci. However, the pattern of variation suggests that most gene flow, although occurring after speciation began, was not recent. There is less evidence of gene flow between D. pseudoobscura and D. p. bogotana. The results are compared with recent work on the genomic locations of genes that contribute to reproductive isolation between D. pseudoobscura and D. persimilis. We show that there is a good correspondence between the genomic regions associated with reproductive isolation and the regions that show little or no evidence of gene flow. PMID- 11919294 TI - Characterization of the intragenomic spread of the human endogenous retrovirus family HERV-W. AB - This study examines the intragenomic spread of the human endogenous retrovirus family HERV-W from insertions present within the draft sequence of the human genome. Identification of shared diagnostic differences and phylogenetic analyses revealed the existence of three main subfamilies. The average divergence between sequences for each of the subfamilies suggests that most of the HERV-W elements were inserted within the genome during a short period of evolutionary time. Each one of the subfamilies consists of two types of insertions, the expected proviral sequences and other sequences resembling the structure of processed retrogenes. These HERV-W retrosequences extend from the R region of the 5' long-terminal repeat (LTR) to the R region of the 3' LTR (as viral genomic RNAs), end in poly(A) 3' tails, and are flanked by direct repeats longer than the proviral integrations. Furthermore, several of the HERV-W retrosequences are 5'-truncated at different sites. I suggest the involvement of the L1 machinery in these integrations and discuss the characteristic features of the evolutionary history of HERV-W, with emphasis on the putative impact of HERV-W retrosequence integrations on the mammalian genome. PMID- 11919295 TI - Efficient biased estimation of evolutionary distances when substitution rates vary across sites. AB - This paper deals with phylogenetic inference when the variability of substitution rates across sites (VRAS) is modeled by a gamma distribution. We show that underestimating VRAS, which results in underestimates for the evolutionary distances between sequences, usually improves the topological accuracy of phylogenetic tree inference by distance-based methods, especially when the molecular clock holds. We propose a method to estimate the gamma shape parameter value which is most suited for tree topology inference, given the sequences at hand. This method is based on the pairwise evolutionary distances between sequences and allows one to reconstruct the phylogeny of a high number of taxa (>1,000). Simulation results show that the topological accuracy is highly improved when using the gamma shape parameter value given by our method, compared with the true (unknown) value which was used to generate the data. Furthermore, when VRAS is high, the topological accuracy of our distance-based method is better than that of a maximum likelihood approach. Finally, a data set of Maoricicada species sequences is analyzed, which confirms the advantage of our method. PMID- 11919296 TI - The complete mitochondrial sequence of Tarsius bancanus: evidence for an extensive nucleotide compositional plasticity of primate mitochondrial DNA. AB - Inconsistencies between phylogenetic interpretations obtained from independent sources of molecular data occasionally hamper the recovery of the true evolutionary history of certain taxa. One prominent example concerns the primate infraordinal relationships. Phylogenetic analyses based on nuclear DNA sequences traditionally represent Tarsius as a sister group to anthropoids. In contrast, mitochondrial DNA (mtDNA) data only marginally support this affiliation or even exclude Tarsius from primates. Two possible scenarios might cause this conflict: a period of adaptive molecular evolution or a shift in the nucleotide composition of higher primate mtDNAs through directional mutation pressure. To test these options, the entire mt genome of Tarsius bancanus was sequenced and compared with mtDNA of representatives of all major primate groups and mammals. Phylogenetic reconstructions at both the amino acid (AA) and DNA level of the protein-coding genes led to faulty tree topologies depending on the algorithms used for reconstruction. We propose that these artifactual affiliations rather reflect the nucleotide compositional similarity than phylogenetic relatedness and favor the directional mutation pressure hypothesis because: (1) the overall nucleotide composition changes dramatically on the lineage leading to higher primates at both silent and nonsilent sites, and (2) a highly significant correlation exists between codon usage and the nucleotide composition at the third, silent codon position. Comparisons of mt genes with mt pseudogenes that presumably transferred to the nucleus before the directional mutation pressure took place indicate that the ancestral DNA composition is retained in the relatively fossilized mtDNA-like sequences, and that the directed acceleration of the substitution rate in higher primates is restricted to mtDNA. PMID- 11919297 TI - A comprehensive vertebrate phylogeny using vector representations of protein sequences from whole genomes. AB - We recently developed a method for producing comprehensive gene and species phylogenies from unaligned whole genome data using singular value decomposition (SVD) to analyze character string frequencies. This work provides an integrated gene and species phylogeny for 64 vertebrate mitochondrial genomes composed of 832 total proteins. In addition, to provide a theoretical basis for the method, we present a graphical interpretation of both the original frequency matrix and the SVD-derived matrix. These large matrices describe high-dimensional Euclidean spaces within which biomolecular sequences can be uniquely represented as vectors. In particular, the SVD-derived vector space describes each protein relative to a restricted set of newly defined, independent axes, each of which represents a novel form of conserved motif, termed a correlated peptide motif. A quantitative comparison of the relative orientations of protein vectors in this space provides accurate and straightforward estimates of sequence similarity, which can in turn be used to produce comprehensive gene trees. Alternatively, the vector representations of genes from individual species can be summed, allowing species trees to be produced. PMID- 11919298 TI - Microsatellite analysis of Drosophila melanogaster populations along a microclimatic contrast at lower Nahel Oren canyon, Mount Carmel, Israel. AB - Drosophila melanogaster populations collected at the south-facing slope (SFS) and north-facing slope (NFS) of lower Nahel Oren canyon, Mount Carmel, Israel display significant differences in survival and longevity at temperature, drought, and starvation stresses. Furthermore, significant assortative mating was previously observed between populations of the two slopes. We used a set of 48 microsatellite markers to analyze patterns of genetic differentiation between D. melanogaster populations from both slopes and D. simulans. Consistent with previous reports, we found D. simulans to be well differentiated from D. melanogaster. Genetic differentiation between SFS and NFS D. melanogaster populations was low (F(ST) = 0.0012). Also a tree of individuals based on the proportion of shared alleles and a model-based clustering method provided no evidence for population substructuring. PMID- 11919299 TI - The crystal structure of eEF1A refines the functional predictions of an evolutionary analysis of rate changes among elongation factors. PMID- 11919300 TI - Accelerated evolution of the surface amino acids in the WD-repeat domain encoded by the hagoromo gene in an explosively speciated lineage of East African cichlid fishes. PMID- 11919301 TI - Two divergent species of the virilis group, Drosophila littoralis and Drosophila virilis, share a replacement polymorphism at the fused locus. PMID- 11919302 TI - Erratic evolution of SRY in higher primates. PMID- 11919303 TI - Reperfusion therapy in acute myocardial infarction. PMID- 11919304 TI - Comparison of angioplasty with stenting, with or without abciximab, in acute myocardial infarction. AB - BACKGROUND: As compared with thrombolytic therapy, primary percutaneous transluminal coronary angioplasty (PTCA) in acute myocardial infarction reduces the rates of death, reinfarction, and stroke, but recurrent ischemia, restenosis, and reocclusion of the infarct-related artery remain problematic. When used in combination with PTCA, coronary stenting and platelet glycoprotein IIb/IIIa inhibitors may further improve outcomes. METHODS: Using a 2-by-2 factorial design, we randomly assigned 2082 patients with acute myocardial infarction to undergo PTCA alone (518 patients), PTCA plus abciximab therapy (528), stenting alone with the MultiLink stent (512), or stenting plus abciximab therapy (524). RESULTS: Normal flow was restored in the target vessel in 94.5 to 96.9 percent of patients and did not vary according to the reperfusion strategy. At six months, the primary end point - a composite of death, reinfarction, disabling stroke, and ischemia-driven revascularization of the target vessel - had occurred in 20.0 percent of patients after PTCA, 16.5 percent after PTCA plus abciximab, 11.5 percent after stenting, and 10.2 percent after stenting plus abciximab (P<0.001). There were no significant differences among the groups in the rates of death, stroke, or reinfarction; the difference in the incidence of the primary end point was due entirely to differences in the rates of target-vessel revascularization (ranging from 15.7 percent after PTCA to 5.2 percent after stenting plus abciximab, P<0.001). The rate of angiographically established restenosis was 40.8 percent after PTCA and 22.2 percent after stenting (P<0.001), and the respective rates of reocclusion of the infarcted-related artery were 11.3 percent and 5.7 percent (P=0.01), both independent of abciximab use. CONCLUSIONS: At experienced centers, stent implantation (with or without abciximab therapy) should be considered the routine reperfusion strategy. PMID- 11919305 TI - Estrogen-receptor polymorphisms and effects of estrogen replacement on high density lipoprotein cholesterol in women with coronary disease. AB - BACKGROUND: Sequence variants in the gene encoding estrogen receptor alpha (ER alpha) may modify the effects of hormone-replacement therapy on levels of high density lipoprotein (HDL) cholesterol and other outcomes related to estrogen treatment in postmenopausal women. METHODS: We characterized 309 women with coronary artery disease who were enrolled in the Estrogen Replacement and Atherosclerosis trial with respect to eight previously described and two newly identified ER-alpha polymorphisms, and we examined the association between these polymorphisms and the response of HDL cholesterol and other lipids to treatment with estrogen alone or estrogen plus progestin. RESULTS: After adjustment for age, race, diabetes status, body-mass index, smoking status, alcohol intake, and frequency of exercise, the 18.9 percent of the women who had the IVS1-401 C/C genotype (i.e., with C on both chromosomes in intervening sequence 1 at position 401 before exon 2) had an increase in the HDL cholesterol level with hormone replacement therapy that was more than twice the increase observed in the other women (13.1 mg per deciliter vs. 6.0 mg per deciliter, P for treatment-by genotype interaction = 0.004); this effect was limited to changes in the HDL subfraction 3 (HDL3) (P for interaction=0.04). Similar patterns of response were observed for three other highly linked ER-alpha intron 1 polymorphisms close to the IVS1-401 site (range of P values for interaction = 0.07 to 0.005). The pattern of increased response of HDL cholesterol in women with the IVS1-401 C/C genotype was evident in both the women receiving estrogen and those receiving estrogen plus progestin, was preserved across racial and ethnic groups, and was significant among women who were compliant with the study medication (P<0.001). CONCLUSIONS: Postmenopausal women with coronary disease who have the ER-alpha IVS1-401 C/C genotype, or several other closely related genotypes, have an augmented response of HDL cholesterol to hormone-replacement therapy. PMID- 11919306 TI - Duration of prophylaxis against venous thromboembolism with enoxaparin after surgery for cancer. AB - BACKGROUND: Abdominal surgery for cancer carries a high risk of venous thromboembolism, but the optimal duration of postoperative thromboprophylaxis is unknown. METHODS: We conducted a double-blind, multicenter trial in which patients undergoing planned curative open surgery for abdominal or pelvic cancer received enoxaparin (40 mg subcutaneously) daily for 6 to 10 days and were then randomly assigned to receive either enoxaparin or placebo for another 21 days. Bilateral venography was performed between days 25 and 31, or sooner if symptoms of venous thromboembolism occurred. The primary end point with respect to efficacy was the incidence of venous thromboembolism between days 25 and 31. The primary safety end point was bleeding during the three-week period after randomization. The patients were followed for three months. RESULTS: The intention-to-treat analysis of efficacy included 332 patients. The rates of venous thromboembolism at the end of the double-blind phase were 12.0 percent in the placebo group and 4.8 percent in the enoxaparin group (P=0.02). This difference persisted at three months (13.8 percent vs. 5.5 percent, P=0.01). Three patients in the enoxaparin group and six in the placebo group died within three months after surgery. There were no significant differences in the rates of bleeding or other complications during the double-blind or follow-up periods. CONCLUSIONS: Enoxaparin prophylaxis for four weeks after surgery for abdominal or pelvic cancer is safe and significantly reduces the incidence of venographically demonstrated thrombosis, as compared with enoxaparin prophylaxis for one week. PMID- 11919307 TI - Images in clinical medicine. Bronchial cast. PMID- 11919308 TI - Psychological sequelae of the September 11 terrorist attacks in New York City. AB - BACKGROUND: The scope of the terrorist attacks of September 11, 2001, was unprecedented in the United States. We assessed the prevalence and correlates of acute post-traumatic stress disorder (PTSD) and depression among residents of Manhattan five to eight weeks after the attacks. METHODS: We used random-digit dialing to contact a representative sample of adults living south of 110th Street in Manhattan. Participants were asked about demographic characteristics, exposure to the events of September 11, and psychological symptoms after the attacks. RESULTS: Among 1008 adults interviewed, 7.5 percent reported symptoms consistent with a diagnosis of current PTSD related to the attacks, and 9.7 percent reported symptoms consistent with current depression (with "current" defined as occurring within the previous 30 days). Among respondents who lived south of Canal Street (i.e., near the World Trade Center), the prevalence of PTSD was 20.0 percent. Predictors of PTSD in a multivariate model were Hispanic ethnicity, two or more prior stressors, a panic attack during or shortly after the events, residence south of Canal Street, and loss of possessions due to the events. Predictors of depression were Hispanic ethnicity, two or more prior stressors, a panic attack, a low level of social support, the death of a friend or relative during the attacks, and loss of a job due to the attacks. CONCLUSIONS: There was a substantial burden of acute PTSD and depression in Manhattan after the September 11 attacks. Experiences involving exposure to the attacks were predictors of current PTSD, and losses as a result of the events were predictors of current depression. In the aftermath of terrorist attacks, there may be substantial psychological morbidity in the population. PMID- 11919309 TI - Clinical practice. Acute exacerbations of chronic obstructive pulmonary disease. PMID- 11919310 TI - Immune thrombocytopenic purpura. PMID- 11919311 TI - Case records of the Massachusetts General Hospital. Weekly clinicopathological exercises. Case 10-2002. A 52-year-old woman with recurrent unsteadiness, slurred speech, and fatigue. PMID- 11919313 TI - Should family members be present during cardiopulmonary resuscitation? PMID- 11919312 TI - Individualized hormone-replacement therapy? PMID- 11919314 TI - Left ventricular assist device. PMID- 11919315 TI - Goal-directed therapy for severe sepsis. PMID- 11919316 TI - Protease inhibitors and mortality among children and adolescents infected with HIV-1. PMID- 11919317 TI - An outbreak of primary pneumonic tularemia. PMID- 11919318 TI - Case 38-2001: paraneoplastic encephalomyelitis and sensory ganglioneuropathy. PMID- 11919319 TI - West Nile encephalitis in Massachusetts. PMID- 11919320 TI - Will parity in coverage result in better mental health care? PMID- 11919321 TI - Hepatocytes and endothelial cells: joining forces to conquer development. PMID- 11919322 TI - S-nitrosoglutathione (GSNO) mediates brain response to hypoxia. PMID- 11919323 TI - Regulation of the globin genes. AB - The a- and b-globin gene clusters are subject to several levels of regulation. They are expressed exclusively in the erythroid cells, only during defined periods of development and in a perfectly tuned way, assuring, at any stage of ontogeny, a correct balance in the availability of a- and b-globin chains for hemoglobin assembling. Such a tight control is dependent on regulatory regions of DNA located either in proximity or at great distances from the globin genes in a region characterized by the presence of several DNAse I hypersensitive sites and known as the Locus Control Region. All these sequences exert stimulatory, inhibitory or more complex activities by interacting with transcription factors that bridge these regions of DNA to the RNA polymerase machinery. Many of these factors have now been cloned and the corresponding mouse genes inactivated, shading new light on the metabolic pathways they control. It is increasingly recognized that such factors are organized into hierarchies according to the number of genes and circuits they regulate. Some genes such as GATA-1 and 2 are master regulators that act on large numbers of genes at early stage of differentiation whereas others, like EKLF, stand on the lowest step and control only single or limited number of genes at late stages of differentiation. We will review recent data gathered from expression studies in cell cultures, in transgenic or K.O. murine models as well as from a clinical settings. We will also discuss the development of novel theories on the regulation of the a- and b globin genes and clusters. PMID- 11919324 TI - Adrenal insufficiency and cardiac dysfunction in the preterm infant. PMID- 11919325 TI - SSRIs in pregnancy--are they safe? PMID- 11919326 TI - Impaired urinary cortisol excretion and early cardiopulmonary dysfunction in immature baboons. AB - Early adrenal insufficiency is associated with cardiopulmonary dysfunction in immature infants. Isolated cortisol levels and ACTH stimulation testing may not adequately show ontogeny of postnatal cortisol secretion nor identify at risk infants. Our objectives were 1) to determine postnatal urinary cortisol excretion rate (UCER) from birth to 14 d in immature baboons and 2) to evaluate the relation between UCER and cardiac performance. UCER was assessed via 6-h blocked urine collections from birth to 336 h of age in twenty-one 125-d gestation (term = 185 d) baboons. Urinary cortisol was measured by RIA. Cardiopulmonary parameters were averaged over the same time periods as urine collection. Serial two-dimensional echocardiograms were performed. After 24-h age, a subgroup (n = 8) received up to four doses (0.5-1.0 mg/kg each) of hydrocortisone for refractory hypotension. UCER significantly increased from 0 to 6 h through 66 to 72 h age for non-cortisol-treated infants. Significantly reduced UCER patterns between birth and 24 h were found for animals subsequently requiring cortisol treatment. Cortisol-treated infants had lower mean blood pressure, worse metabolic acidosis, increased fluid needs, and impaired left ventricular function between 12 and 48 h of age. No group differences were found in gas exchange or ventilator support. We conclude that adrenal cortisol secretion significantly improves over the initial 72 h of life in the 125-d immature baboon. Failure to increase UCER after 12-24 h of life correlated with poor cardiovascular function that improved with hydrocortisone therapy. Adrenal hypofunction in the immature baboon is similar to the very preterm human and could serve as a model for future postnatal investigations. PMID- 11919327 TI - Effect of maternal fluoxetine administration on uterine blood flow, fetal blood gas status, and growth. AB - Clinical depression, diagnosed in 5-15% of women during pregnancy, increases the risk of negative pregnancy outcomes including an increased incidence of low birth weight newborns and preterm delivery. Fluoxetine, a selective serotonin reuptake inhibitor, is often prescribed to treat depression due to its efficacy, high margin of safety, and mild side effects. However, fluoxetine initially increases plasma serotonin concentration, and serotonin causes uterine vasoconstriction in sheep, which could result in fetal hypoxemia. To assess fetal fluoxetine effects, late-gestation pregnant sheep were surgically prepared for the measurement of blood gases, heart rate, blood pressure, and uterine artery blood flow (n = 29). Ewes received a 70-mg bolus i.v. infusion of fluoxetine over 2 min in 10 mL of sterile water followed by continuous infusion at a rate of 100 microg/min for 8 d (n = 14), or continuous infusion of sterile water (n = 15). Transient decreases in uterine artery blood flow, fetal PO(2), and oxygen saturation were observed within the first 15 min after fluoxetine exposure, which did not return to normal values by 24 h. Fetal pH decreased and PCO(2) increased over the first 4 h with a return to normal by 24 h. However, there were no differences in uterine artery blood flow, blood gas status, or cardiovascular measures between the control and fluoxetine group over the rest of the 8-d infusion period. Thus, fluoxetine exposure during pregnancy has transient effects on fetal status that may be of developmental consequence if they occur repetitively. PMID- 11919328 TI - Prolonged prenatal psychotropic medication exposure alters neonatal acute pain response. AB - Selective serotonin reuptake inhibitors (SSRIs) and benzodiazepines are frequently used to treat maternal depression during pregnancy, however the effect of increased serotonin (5HT) and gamma-amino-butyric acid (GABA) agonists in the fetal human brain remains unknown. 5HT and GABA are active during fetal neurologic growth and play early roles in pain modulation, therefore, if prolonged prenatal exposure alters neurodevelopment this may become evident in altered neonatal pain responses. To examine biologic and behavioral effects of prenatal exposure, neonatal responses to acute pain (phenylketonuria heel lance) in infants with prolonged prenatal exposure were examined. Facial action (Neonatal Facial Coding System) and cardiac autonomic reactivity derived from the relationship between respiratory activity and short term variations of heart rate (HRV) were compared between 22 infants with SSRI exposure (SE) [fluoxetine (n = 7), paroxetine (n = 11), sertraline (n = 4)]; 16 infants exposed to SSRIs and clonazepam (SE+) [paroxetine (n = 14), fluoxetine (n = 2)]; and 23 nonexposed infants during baseline, lance, and recovery periods of a heel lance. Length of maternal SSRI use did not vary significantly between exposure groups-[mean (range)] SE:SE+ 183 (31-281):141 (54-282) d (p > 0.05). Infants exposed to SE and SE+ displayed significantly less facial activity to heel lance than control infants. Mean HR increased with lance, but was significantly lower in SE infants during recovery. Using measures of HRV and the transfer relationship between heart rate and respiration, SSRI infants had a greater return of parasympathetic cardiac modulation in the recovery period, whereas a sustained sympathetic response continued in the control group. Prolonged prenatal SSRI exposure appears to be associated with reduced behavioral pain responses and increased parasympathetic cardiac modulation in recovery following an acute neonatal noxious event. Possible 5HT-mediated pain inhibition, pharmacologic factors and the developmental course remain to be studied. PMID- 11919329 TI - Auditory neural responses to click stimuli of different rates in the brainstem of very preterm babies at term. AB - Auditory neural responses to acoustic stimuli of different rates were studied by analyzing changes in brainstem auditory evoked responses (BAER) with increasing repetition rate of clicks, or rate-dependent changes, in 62 very preterm babies (gestation 24-32 wk). None had perinatal asphyxia or major complications at the time of testing (37-42 wk postconceptional age) to exclude their possible effects on the recorded BAER. As the rate of clicks was increased from 21/s to 51/s and 91/s, I-V interpeak interval in these babies increased similarly to that in normal term neonates. The rate-dependent change decreased significantly in I-III interval, but increased significantly in III-V intervals and III-V/I-III interval ratio (all p < 0.01). At all three rates of clicks, the I-V interval, the most commonly used BAER variable, tended to increase slightly but did not differ significantly from the term neonates. The I-III interval decreased significantly at higher click rates (ANOVA p < 0.05 at 51/s and < 0.001 at 91/s), whereas the III-V interval and III-V/I-III interval ratio increased significantly at all 21/s, 51/s, and particularly 91/s (p < 0.01-0.001). No abnormalities were found in wave V amplitude at any rates. These results suggest that very preterm babies have an advanced peripheral development of the brainstem auditory pathway but a retarded central development or central impairment. A nearly normal I-V interval does not exclude a possible abnormality in its subcomponents (I-III and III-V intervals). In babies who have a normal BAER at a conventionally used low rate of clicks, we cannot exclude an abnormal BAER at higher rates. PMID- 11919330 TI - Effect of multisensory stimulation on analgesia in term neonates: a randomized controlled trial. AB - Many attempts have been made to obtain safe and effective analgesia in newborns. Oral glucose-water has been found to have analgesic properties in neonates. We investigated whether other sensory stimulation added to oral glucose provided more effective analgesia than oral glucose alone. In a randomized prospective double-blind trial, we studied 120 term newborns during heel prick. The babies were divided randomly into six groups of 20, and each group was treated with a different procedure during heel prick: A) control; B) 1 mL 33% oral glucose given 2 min before the heel prick; C) sucking; D) 1 mL 33% oral glucose plus sucking; E) multisensory stimulation including 1 mL 33% oral glucose (sensorial saturation); F) multisensory stimulation without oral glucose. Sensorial saturation consisted in massage, voice, eye contact, and perfume smelling during heel prick. Each heel prick was filmed and assigned a point score according to the Douleur Aigue du Nouveau-ne (DAN) neonatal acute pain scale. Camera recording began 30 s before the heel prick, so it was impossible for the scorers to distinguish procedure A (control) from B (glucose given 2 min before), C (sucking water) from D (sucking glucose), and E (multisensory stimulation and glucose) from F (multisensory stimulation and water) from the video. Procedure E (multisensory stimulation and glucose) was found to be the most effective procedure, and the analgesia was even more effective than that produced by procedure D (sucking glucose). We conclude that sensorial saturation is an effective analgesic technique that potentiates the analgesic effect of oral sugar. It can be used for minor painful procedures on newborns. PMID- 11919331 TI - Ovine placental and fetal arginine metabolism at normal and increased maternal plasma arginine concentrations. AB - Arginine (A) may play a significant role in fetal growth, by stimulating insulin secretion and as a precursor for both polyamine synthesis and nitric oxide production. To determine whether increased maternal plasma A concentrations can enhance delivery of A to the fetus, uterine, umbilical, and net uteroplacental (UP) A uptake rates were measured in 12 pregnant ewes at 129.6 +/- 0.4 d gestation (mean +/- SEM) during normal and after 3 h of increased maternal plasma A concentrations. With a 2.7-fold increase in maternal plasma A concentrations (p < 0.001), there were significant increases in uterine A uptake (13.8 +/- 1.0 to 41.3 +/- 7.7 micromol/min, p < 0.005), umbilical A uptake (3.3 +/- 0.5 to 5.2 +/- 0.8 micromol.min(-1).kg(-1) fetus, p < 0.005), UP A uptake (17.8 +/- 6.2 to 89.2 +/- 20.3 micromol.min(-1).kg(-1) placenta, p < 0.01), fetal arterial A concentration (98.7 +/- 6.3 to 137.1 +/- 9.9 microM, p < 0.001), maternal A disposal rate (143.7 +/- 9.4 to 217.0 +/- 6.7 micromol/min, p < 0.001), fetal A disposal rate (7.9 +/- 0.8 to 9.9 +/- 1.1 micromol.min(-1).kg(-1), p < 0.05), fetal A oxidation rate (1.31 +/- 0.24 to 1.84 +/- 0.36 micromol.min(-1).kg(-1), p < 0.05), and plasma insulin concentration in both fetus (16 +/- 2 to 20 +/- 2 microU/mL, p < 0.001) and mother (24 +/- 3 to 32 +/- 4 microU/mL, p < 0.001). Thus, increased maternal plasma A concentration increases maternal, UP, and fetal A net uptake, and increases insulin secretion in mother and fetus. The 4.2-fold larger increase in UP than net fetal A uptake could represent preferential UP A metabolism relative to fetal A metabolism, relatively limited placental-fetal A transport capacity compared with uterine A uptake capacity, or both; responsible mechanisms remain unknown. PMID- 11919333 TI - Differential diagnosis between Pendred and pseudo-Pendred syndromes: clinical, radiologic, and molecular studies. AB - The disease gene for Pendred syndrome has been recently characterized and named PDS. It codes for a transmembrane protein called pendrin, which is highly expressed at the apical surface of the thyroid cell and functions as a transporter of chloride and iodide. Pendrin is also expressed at the inner ear level, where it appears to be involved in the maintenance of the endolymph homeostasis in the membranous labyrinth, and in the kidney, where it mediates chloride-formate exchange and bicarbonate secretion. Mutations in the PDS gene and the consequent impaired function of pendrin leads to the classic phenotype of Pendred syndrome, i.e. dyshormonogenic goiter and congenital sensorineural hearing loss. In the present study, we performed a detailed clinical, radiologic, and molecular analysis of six families presenting with clinical diagnosis of Pendred syndrome. In two families a homozygous pattern for PDS mutations was found, whereas the affected members of the other four families were compound heterozygotes. One family did not harbor PDS mutations. Among the four novel mutations described, one is a transversion in exon 2 (84C>A), leading to the substitution S28R. Two other novel mutations lie in exon 4 (398T>A) and in exon 16 (1790T>C), leading to the substitutions S133T and L597S, respectively. The fourth novel mutation (1614+1G>A) is located in the first base pair of intron 14, probably affecting the splicing of the PDS gene. Clinically, all patients had goiter with positive perchlorate test, hypothyroidism, and severe or profound sensorineural hearing loss. In all the individuals harboring PDS mutations, but not in the family without PDS mutations, inner ear malformations, such as enlargement of the vestibular aqueduct and of the endolymphatic duct and sac, were documented. The pseudo-Pendred phenotype exhibited by the family without PDS mutations is likely caused by an autoimmune thyroid disease associated with a sensorineural hearing loss of different origin. PMID- 11919332 TI - Erythropoietin stimulates vasculogenesis in neonatal rat mesenteric microvascular endothelial cells. AB - Human breast milk is a rich source of growth factors, including erythropoietin (Epo), the endogenous hormonal stimulant of erythropoiesis. Recombinant human Epo (rhEpo) has been shown to stimulate 1) angiogenesis, the process of new blood vessel growth from preexisting vessels; 2) vasculogenesis, tubule formation from single-cell suspensions; and 3) endothelial cell proliferation in immortalized endothelial cells and vessel explants. We hypothesized that Epo would induce mitogenesis and stimulate vasculogenesis in primary cultures of microvascular endothelial cells (MVECs) from neonatal rat mesentery. Isolation, purification, characterization, and culture of MVECs were performed. Cell proliferative effects of rhEpo were studied by 3-(4,5 dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay in cultured MVECs. Vasculogenic effects of rhEpo were examined on cultured MVECs plated on either hormone-rich Matrigel substratum or the extracellular matrix protein, type I collagen. Our findings show that MVECs are isolated and purified, and that rhEpo stimulates MVEC proliferation, with maximal proliferation seen with a concentration of 50 IU/mL rhEpo. Tubule formation assays reveal that an rhEpo concentration of 50 IU/mL produces maximal tubule formation after 12 h on both Matrigel and the simple substratum, type I collagen. Our study is the first to examine the effects of rhEpo on the endothelium of the neonatal gastrointestinal tract. These data suggest that Epo may have a trophic effect on the vasculature of the gastrointestinal tract early in development. Furthermore, as Epo has been measured in breast milk, and its receptor has been shown to exist on the mucosa and gastrointestinal vasculature, Epo may be an endogenous stimulant of vessel growth during neonatal gastrointestinal development. PMID- 11919334 TI - Vasodilation to vascular endothelial growth factor in the uterine artery of the pregnant rat is blunted by low dietary protein intake. AB - Pregnancy is associated with a substantial increase in uterine artery blood flow, which may in part result from dilation in response to vascular endothelial growth factor (VEGF). Uterine blood flow is reported to be reduced in globally diet restricted pregnant rats. Both global and protein dietary restriction in pregnancy produce programmed effects in offspring. In this study we hypothesized that protein restriction in pregnancy impairs maternal uterine artery responses to VEGF. Vascular responses to VEGF were determined in isolated uterine arteries of pregnant (18 or 19 d of gestation) Wistar rats fed a diet containing either 18% or 9% casein throughout pregnancy. For comparison, responses to phenylephrine, potassium chloride, and acetylcholine were determined. In addition, the response of the mesenteric artery to VEGF was studied in the same animals. A significant reduction of the maximal relaxation to VEGF (p = 0.041) and in the overall response (p = 0.004) to VEGF was found in uterine arteries of the 9% compared with the 18% group, but responses to all other agonists were similar. The VEGF response was reduced by cyclooxygenase inhibition (indomethacin) in both groups. In the 18%, but not the 9%, group it was further reduced by nitric oxide synthase inhibition (Nomega-nitro-L-arginine methyl ester). VEGF was shown to dilate the mesenteric artery but this effect was not significantly altered by the low-protein diet. These results show an attenuated uterine artery vasodilator response to VEGF produced by a low-protein diet in pregnancy, partly because of a reduction of the nitric oxide component of VEGF mediated relaxation. PMID- 11919335 TI - Expression in the placenta of neuronal markers for perinatal brain damage. AB - Determination of S-100 a and b and neuron-specific enolase (NSE) in (cord) blood and amniotic fluid has been used to assess neonatal neuronal damage after compromising conditions. However, these proteins are not only found in nervous tissue, and their expression in placenta and umbilical cord has never been investigated. In this study, S-100 (a and b) and NSE expression in human cord and placental tissue was studied by immunohistochemical analysis. Similar analysis was performed using two other brain-specific markers: glial fibrillary acidic protein and growth-associated protein B-50 (also known as GAP-43 or neuromodulin). Tissue was derived after elective cesarean section in seven women of different gestational ages after uncomplicated or complicated pregnancy. S-100 a and b and NSE immunoreactivity was found in several cell types and structures in the umbilical cord as well as in the placenta of all seven cases. Glial fibrillary acidic protein and B-50 showed no immunoreactivity. These data are of importance for interpreting findings of studies in which S-100 or NSE levels in cord blood or amniotic fluid have been related to neuronal damage in the neonate. The increased levels found may just as well be caused by leakage from placenta or umbilical cord as be caused by brain damage. We conclude that S-100 a and b and NSE are not suitable markers for neonatal brain damage. Brain-restricted proteins such as glial fibrillary acidic protein and B-50 seem more promising. PMID- 11919336 TI - Clinical and biochemical determinants of bone metabolism and bone mass in adolescent female patients with anorexia nervosa. AB - Among pathologies prevalent in western societies, anorexia nervosa has increased over the last decade. Its effects on bone mass need to be defined, and prognostic factors, either clinical or biochemical, could aid clinicians in individual patient management. To determine which clinical and/or biochemical parameters could be related to bone mass status in adolescent female anorexia nervosa patients, 73 female patients were classified according to different stages of their illness and studied in terms of clinical and biochemical parameters and bone densitometric mineral content at lumbar spine. Patients (age 17.2 +/- 1.7 y, mean +/- SD) with Tanner pubertal stage 5, regular menstruation for more than 3 mo before the onset of secondary amenorrhea, and diagnosed with anorexia nervosa were consecutively studied and classified in three clinical situations: I) active phase (34 patients): undernourished and amenorrheic; II) weight recovered but still amenorrheic (20 patients); III) fully recovered (19 patients). Clinical data were recorded at the time of bone density measurement, concomitant with blood sample extraction for study of IGF-I, IGF-binding protein 3 (IGFBP-3), IGFBP-1, estradiol, sex hormone-binding globulin, dehydroepiandrosterone sulfate, prealbumin, amino-terminal propeptide of procollagen III, osteocalcin, bone alkaline phosphatase, carboxy-terminal propeptide of procollagen I, amino terminal propeptide of procollagen I, carboxy-terminal telopeptide of collagen I, 25-OH-vitamin D, 1,25(OH)(2)-vitamin D, and parathormone. In addition, a 24-h urine collection was made for cortisol, GH, deoxypyridinoline, amino-terminal telopeptide of collagen I, and calcium and creatinine content analysis. IGF-I, estradiol, and biochemical bone formation markers were higher and IGFBP-1, sex hormone-binding globulin, and biochemical bone resorption markers were lower in the weight-recovered stages (stages II and III) compared with the active phase (stage I). Bone formation markers correlated positively with body mass index SD score and IGF-I, whereas bone resorption markers correlated negatively with body mass index SD score and estradiol. Although no statistically significant differences regarding lumbar spine bone mineral density SD score values were recorded among the three stages of the illness, the proportion of osteopenic patients was clearly lower among stage III patients. The actual bone mineral density was inversely related to the duration of amenorrhea and directly related to duration of postmenarcheal menses before amenorrhea. In addition, a subset of osteopenic patients (five of 19) in the fully clinically recovered group with accelerated bone turnover was identified. Normal circulating estrogen level exposure time predicts actual bone mineral density at lumbar spine in young adolescent anorexia nervosa patients. In addition to psychiatric and nutritional interventions, estrogen-deprivation periods must be shortened to less than 20 mo. Patients remaining osteopenic at full clinical recovery require additional follow up studies. PMID- 11919337 TI - Muscle analysis by measurement of maximal isometric grip force: new reference data and clinical applications in pediatrics. AB - Skeletal muscle development is one of the key features of childhood and adolescence. Determining maximal isometric grip force (MIGF) using a hand-held Jamar dynamometer is a simple method to quantify one aspect of muscle function. Presently available reference data present MIGF as a function of chronological age. However, muscle force is largely determined by body size, and many children undergoing muscle performance tests in the clinical setting suffer from growth retardation secondary to a chronic disorder. Reference data were established from simple regressions between age or log height and log MIGF in a population of 315 healthy children and adolescents aged 6 to 19 y (157 girls). These data were used to calculate age- or height-dependent SD scores (SDS) for MIGF in three pediatric patient groups. In renal graft recipients (n = 14), the age-dependent MIGF SDS was markedly decreased (-2.5 +/- 1.9; mean +/- SD). However, these patients had short stature (height SDS, -2.5 +/- 1.2), and the height-dependent MIGF SDS was close to normal (-0.4 +/- 1.5). Similarly, in cystic fibrosis patients (n = 13) age-dependent MIGF SDS was -1.6 +/- 1.6, but height-dependent MIGF SDS was -0.5 +/- 1.1. Children with epilepsy who were taking anticonvulsant therapy (n = 34) had normal stature, and consequently age- and height-dependent MIGF SDS were similar (0.4 +/- 1.0 and 0.4 +/- 0.8, respectively). In conclusion, MIGF determination provides information on an important aspect of physical development. Height should be taken into account to avoid misinterpretation. PMID- 11919338 TI - Mutations at the galactose-1-p-uridyltransferase gene in infants with a positive galactosemia newborn screening test. AB - Newborn screening for galactosemia yields a high number of false-positive results. Confirmatory DNA testing for unknown galactosemia mutations on the initial positive sample using novel techniques of mutation detection tenders itself to reduce the recall rate. The potential benefits of confirmatory DNA testing, however, could be offset by the detection of a high percentage of galactosemia carriers, Duarte/galactosemia compound heterozygotes, and infants with benign sequence changes in the galactose-1-phosphate uridyltransferase (GALT) gene among infants with a positive biochemical screening test. Our aim was to determine the frequency and allelic distribution of all sequence changes in the GALT gene in 110 newborns with a positive total galactose screening test among 43,688 Austrian newborns screened consecutively. We found that only 20 of the 110 probands carried at least one known or novel candidate galactosemia mutation (one galactosemia homozygote, 7 Duarte/galactosemia compounds, 12 carriers) as judged by denaturing gradient gel electrophoresis and cleavage fragment length polymorphism analysis. Four novel galactosemia candidate mutations (Q9H, A46fsdelCAGCT, M129T, L342I) were identified. Sixty-seven probands had no detectable sequence changes and 23 carried only the benign Duarte or Los Angeles variant alleles or silent mutations. We conclude that a rapid and automatable confirmation test for unknown GALT mutations, e.g. on a high-density oligonucleotide array basis, has the potential to lower the recall rate of galactosemia screening in our population by about five-fold from 0.25 to 0.046%. Further research, however, will be required before the development of such a test can be advocated. PMID- 11919339 TI - Lubeluzole pretreatment does not provide neuroprotection against transient global cerebral ischemia in fetal sheep near term. AB - The aim of the present study was to test the neuroprotective effect of the novel benzothiazol compound lubeluzole on neuronal cell damage in fetal sheep arising from global cerebral ischemia. Thirteen fetal sheep were prepared at a mean gestational age of 127 +/- 1 d (term is at 147 d). Six fetuses were treated with lubeluzole (0.33 mg/kg estimated body weight) before induction of global cerebral ischemia (-90, -60, and -30 min), while the remainder (n = 7) received solvent. Cerebral ischemia was induced by occluding both carotid arteries for 30 min. Cerebral blood flow was measured by injecting radio-labeled microspheres before ( 90 min), during (+3 min and +27 min), and after (+40 min, +3 h, and +72 h) cerebral ischemia. Neuronal cell damage was assessed in the cerebrum and deeper brain structures by light microscopy. Values are given as means +/- SD. In control fetuses, blood flow to the cerebrum was reduced from 100 +/- 25 mL.100 g( 1) min(-1) to less than 20 mL.100 g(-1) min(-1) during ischemia. Shortly after ischemia, hyperperfusion occurred (217 +/- 66 mL.100 g(-1)min(-1)) followed by a tendency toward hypoperfusion (72 +/- 17 mL.100 g(-1) min(-1)) later on (+3 h). Significant differences in blood flow to the various brain structures between the control and study groups could not be observed. Neuronal cell damage was concentrated in the parasagittal regions of the cerebrum. Preischemic application of lubeluzole did not have any effect on the extent of neuronal cell damage. From these results, we conclude that pretreatment with lubeluzole fails to protect the brain of fetal sheep near term from injury after transient global cerebral ischemia. However, because the observation period lasted only 3 d, a possible effect of lubeluzole on pathophysiological mechanisms inducing delayed neuronal cell death cannot be fully excluded. PMID- 11919340 TI - Effects of active compression-decompression cardiopulmonary resuscitation with the inspiratory threshold valve in a young porcine model of cardiac arrest. AB - Active compression-decompression (ACD) cardiopulmonary resuscitation (CPR) with the inspiratory threshold valve (ITV) has been recently recommended by the American Heart Association for treatment of adults in cardiac arrest (class IIb: alternative, useful intervention), but this new technique has never been used in a pediatric population. Thus, this study was designed to evaluate ACD + ITV CPR in a young porcine model of cardiac arrest. After 10 min of ventricular fibrillation, and 8 min of standard CPR, ACD + ITV CPR was performed in seven 4- to 6-wk-old pigs (8-12 kg); defibrillation was attempted 8 min later. Within 2 min after initiation of ACD + ITV CPR, mean (+/- SEM) coronary perfusion pressure increased from 18 +/- 2 to 24 +/- 3 mm Hg (p = 0.018). During standard versus ACD + ITV CPR, mean left ventricular myocardial and total cerebral blood flow was 59 +/- 21 versus 126 +/- 32 mL.min(-1).100 g(-1), and 36 +/- 7 versus 60 +/- 15 mL.min(-1).100 g(-1), respectively (p = 0.028). Six of seven animals were successfully defibrillated, and survived >15 min. In conclusion, the combination of ACD + ITV CPR significantly increased both coronary perfusion pressure and vital organ blood flow after prolonged standard CPR in this young porcine model of ventricular fibrillation. PMID- 11919341 TI - Saccharomyces boulardii enhances N-terminal peptide hydrolysis in suckling rat small intestine by endoluminal release of a zinc-binding metalloprotease. AB - Saccharomyces boulardii (S. boulardii), a biotherapeutic agent effective in acute and chronic enterocolopathies, produces trophic intestinal effects at least in part mediated by the endoluminal release of polyamines. However, the effects of the yeast on peptide hydrolysis have not yet been studied. The objectives of this study were to assess in suckling rats the endoluminal and mucosal aminopeptidase activities in response to S. boulardii treatment and to analyze their related mechanisms. Peptidase activities were assayed on yeast cells by using several L amino acid-p-nitroanilide substrates in the pH range of 2 to 10. A marked hydrolytic activity was found for L-leucine-p-nitroanilide that peaked at pH = 8 (K(m) = 0.334 mM, V(max) = 44.7 micromol.min(-1).g(-1) protein). N-terminal peptide hydrolysis was confirmed using as substrate L-Leu-Gly-Gly (K(m) = 4.71 mM, V(max) = 18.08 micromol.min(-1).g(-1) protein). Enzyme reactions were inhibited in the presence of 1 mM Zn(2+). Oral treatment of sucklings with S. boulardii significantly enhanced jejunal and ileal mucosal leucine-aminopeptidase activities by 24 and 34%, respectively, over controls. In concordance, aminopeptidase activity was enhanced in jejunal and ileal endoluminal fluid samples by 47 and 105%, respectively. By use of an IgG-purified antibody raised against the zinc-binding domain common to metalloproteases, the yeast aminopeptidase was immunoprecipitated and detected as an heteromeric enzyme of 108 and 87-kD subunits. S. boulardii, when given orally to suckling rats, is able to significantly enhance hydrolysis of N-terminal oligopeptides in both endoluminal fluid and intestinal mucosa by the endoluminal release of a leucine aminopeptidase that appears to be a zinc-binding metalloprotease belonging to the M1 family of peptidases. PMID- 11919342 TI - Rat cultured neuronal and glial cells respond differently to toxicity of unconjugated bilirubin. AB - High levels of unconjugated bilirubin (UCB) can be neurotoxic. Nevertheless, the mechanism of UCB interaction with neural cells is still unknown. This study investigates whether cultured rat neurons and astrocytes respond differently to UCB exposure. UCB toxicity was evaluated by lactate dehydrogenase release, induction of apoptosis, cytoskeleton degeneration, 3-(4,5-dimethylthiazol-2-yl) 2,5-diphenyltetrazolium bromide (MTT) reduction, and glutamate uptake. Primary cultures of rat brain astrocytes and neurons were incubated at 37 degrees C with 85.5 microM UCB plus 28.5 microM albumin for 4 h. In assays of glutamate uptake, cells were exposed to 80-120 microM UCB plus 100 microM albumin for 15 min. The results showed that after incubation with 85.5 microM UCB, lactate dehydrogenase release was greater in neurons than in astrocytes (38% versus 14%, p < 0.05). Also, levels of apoptosis were markedly enhanced in neurons (29% versus 19%, p < 0.01). In accordance, neuronal cytoskeleton disassembly was evident during incubation with 85.5 microM UCB, whereas equivalent effects on astrocytes required as much as 171 microM. Conversely, inhibition of MTT metabolism and glutamate uptake by UCB was more pronounced in astrocytes than in neurons (74% versus 60%, p < 0.05 and 41% to 56% versus 25% to 33%, p < 0.05, respectively). In conclusion, the study demonstrates that astrocytes are more susceptible to inhibition of glutamate uptake and MTT reduction by UCB, whereas neurons are more sensitive to cell death by necrosis or apoptosis. These results suggest that UCB is toxic to both astrocytes and neurons, although through distinct pathways. PMID- 11919343 TI - American Pediatric Society John Howland Award 2001: presentation and acceptance. PMID- 11919345 TI - Getting in the game. PMID- 11919347 TI - Consumer & family information: hypochondriasis. PMID- 11919348 TI - Datapoints: trends in research fellowship opportunities in psychiatry. PMID- 11919349 TI - Law & psychiatry: policing expert testimony: the role of professional organizations. PMID- 11919350 TI - State mental health policy: critical elements of public-sector managed behavioral health programs for severe mental illness in five States. PMID- 11919351 TI - Managed care: strengthening the consumer voice in managed care: IV. The Leadership Academy Program. PMID- 11919352 TI - Economic grand rounds: economics and the Surgeon General's Report on Mental Health. PMID- 11919353 TI - Personal accounts: life as death: hope regained with ECT. PMID- 11919354 TI - Articles on special populations published in Psychiatric Services between 1950 and 1999. AB - OBJECTIVE: The authors sought to determine the number of articles on special populations published in this journal between 1950 and 1999 and the specific populations addressed. METHODS: The authors reviewed original articles, review articles, brief reports, and columns published in this journal between 1950 and 1999. They tallied articles on the mentally retarded population, women, children and adolescents, elderly persons, gays and lesbians, and parents and families. Articles that addressed patients' communities and villages were also counted, as were articles in the multicultural category-those that addressed more than one racial or ethnic group. RESULTS: Between 1950 and 1999, 578 issues of the journal were published, containing approximately 7,976 articles. A total of 555 of articles (7 percent) addressed special populations. More than two-thirds of these articles addressed the mental health issues of elderly persons (178 articles) and of children and adolescents (200 articles). Thirty-eight of the 555 articles (7 percent) addressed women's issues. A total of 120 articles (22 percent) addressed one or more racial or ethnic group; the largest proportion-36 articles-focused on African Americans, 21 addressed issues of Latinos, and 27 had a multicultural focus. Only two articles published during the 50-year period addressed mental health issues of gays and lesbians, and only five focused on the mentally retarded population. CONCLUSIONS: The journal has made continuous progress in increasing the number of articles on special populations. Editorial policies can help encourage submission and publication of articles on these groups. PMID- 11919355 TI - Prescription of antipsychotic drugs by office-based physicians in the United States, 1989-1997. AB - OBJECTIVE: This study examined trends in the prescription of antipsychotic drugs in a nationally representative sample of physicians in nonfederal office-based clinical practice during the 1990s. METHODS: The authors analyzed physician reported data from annual National Ambulatory Medical Care Surveys between 1989 and 1997 using weighted national estimates of physician visits during which antipsychotic drugs were prescribed. Prescription rates for antipsychotic drugs were compared between periods and among demographic, organizational, and clinical subgroups. RESULTS: Prescription of antipsychotic drugs in office-based practice increased significantly between 1989 and 1997. In 1989 antipsychotics were prescribed during 3.2 million office visits (.46 percent of all visits), compared with 6.9 million visits in 1997 (.88 percent). The atypical antipsychotics risperidone and olanzapine were the most widely prescribed antipsychotics in 1997. Risperidone was prescribed during 22.8 percent of all visits that involved prescription of an antipsychotic, and olanzapine during 17.1 percent. Psychiatrists were more likely than other physicians to prescribe an atypical agent (37.1 percent of visits involving prescription of an antipsychotic compared with 14.2 percent). Psychiatrists were also more likely than other physicians to schedule a follow-up visit after prescribing an antipsychotic (96.6 percent of visits compared with 73 percent). No evidence was found of a broadening of diagnostic indications for use over time. CONCLUSIONS: The rate of prescription of antipsychotic drugs among office-based physicians increased sharply during the 1990s after a nine-year decline. The increase was accounted for by growth in the use of atypical antipsychotics; the overall prescription rate of conventional agents did not change. Psychiatrists were more likely to prescribe atypical agents and to monitor more closely patients who were taking antipsychotics. PMID- 11919356 TI - A model for managed behavioral health care in an academic department of psychiatry. AB - In response to the effects of the managed care environment on patient flow and care, the department of psychiatry and behavioral sciences of the Johns Hopkins School of Medicine developed and has been managing a capitated behavioral health care program. The program is responsible for providing mental health and substance abuse services for 22,000 members of the TRICARE Uniformed Services Family Health Plan (USFHP), directed by the U.S. Department of Defense. The integration of primary care and behavioral health care is a major feature of the USFHP/TRICARE program. The authors describe the transition from a carve-out for profit managed care organization to the integrated program managed by the department. During the first two years of the program, access to services increased and use of inpatient services decreased without the need to deny service use. To supplement previous reports of the involvement of academic psychiatry departments in behavioral health care, the authors supply utilization and financial data that may serve as benchmarks for similar efforts by other departments. PMID- 11919357 TI - Outcomes and service use among homeless persons with serious mental illness and substance abuse. AB - OBJECTIVE: This study compared baseline characteristics and clinical improvement after 12 months among homeless persons with a diagnosis of serious mental illness with and without a comorbid substance use disorder. METHODS: The study subjects were 5,432 homeless persons with mental illness who were participating in the Center for Mental Health Services' Access to Community Care and Effective Services and Supports (ACCESS) program. Analysis of covariance was used to compare clients who had dual diagnoses and those who did not and to identify any association between service use and clinical improvement. RESULTS: Follow-up data were available for 4,415 clients (81 percent). At baseline, clients with dual diagnoses were worse off than those without dual diagnoses on most clinical and social adjustment measures. Clients with dual diagnoses also had poorer outcomes at follow-up on 15 (62 percent) of 24 outcome measures. However, among clients with dual diagnoses, those who reported extensive participation in substance abuse treatment showed clinical improvement comparable to or better than that of clients without dual diagnoses. On measures of alcohol problems, clients with dual diagnoses who had a high rate of participation in self-help groups had outcomes superior to those of other clients with dual diagnoses. Clients with dual diagnoses who received high levels of professional services also had superior outcomes in terms of social support and involvement in the criminal justice system. CONCLUSIONS: Homeless persons with dual diagnoses had poorer adjustment on most baseline measures and experienced significantly less clinical improvement than those without dual diagnoses. However, those with dual diagnoses who received extensive substance abuse treatment showed improvement similar to those without at 12 months. PMID- 11919358 TI - Use of a state inpatient forensic system under managed mental health care. AB - OBJECTIVES: One of the goals of managed mental health care has been to lower the use of inpatient psychiatric treatment. In the past, interventions that have limited hospitalization for persons with severe mental illness have led to greater involvement of these individuals with the criminal justice and forensic mental health systems. The authors examined associations between Medicaid managed mental health care in Massachusetts and rates of admission to the inpatient forensic mental health service maintained by the state's mental health department. METHODS: A total of 7,996 persons who were receiving services from the department before and after the introduction of managed care were studied. A logistic regression model based on generalized estimating equations was used to identify associations between Medicaid beneficiary status and forensic hospitalization before and after the introduction of managed care. RESULTS: The overall rate of forensic hospitalization declined in the study cohort in both periods. However, no significant decline was observed in the risk of forensic hospitalization among Medicaid beneficiaries whose care had become managed. CONCLUSIONS: Although the results of this study warrant further exploration, the risk of forensic hospitalization among Medicaid beneficiaries should be considered by policy makers in the design of mental health system interventions. PMID- 11919359 TI - Identifying meaningful subgroups of adults with severe mental illness. AB - OBJECTIVE: The authors describe work conducted to identify, describe, and validate subgroups, or clusters, of adults with severe mental illness. METHODS: Clusters were first identified by using functional assessment ratings and statistical clustering procedures. Seven separate cluster analyses were accomplished, involving agencies in 15 counties in Ohio. Brief prose descriptions of the clusters were developed on the basis of the results of the statistical analyses. These descriptions were then used as stimuli in an extended expert based validation and enhancement process that included work groups of consumers, family members, and providers. RESULTS: This sequential process produced generalizable, holistic descriptions of five core clusters of individuals with common strengths, problems, treatment histories, and life situations: older people in poor health who have psychiatric symptoms; individuals who have both psychiatric and substance use disorders; chronically mentally ill persons who have long treatment histories and severe disabilities; persons who appear to function well in the community but who are isolated as a result of anxiety, depression, or trauma-related social fears; and mental health consumers who function well in the community and who use personal and professional support systems to manage their mental health problems. CONCLUSIONS: The results support the hypothesis that there is meaningful, but predictable, heterogeneity among persons who have severe mental illness. The clusters identified in this study can be used to plan, manage, and evaluate services. PMID- 11919360 TI - Status of patients with first-episode psychosis after one year of phase-specific community-oriented treatment. AB - OBJECTIVE: A phase-specific intervention provided soon after the onset of a first episode of psychosis is likely to engender a more hopeful outlook. This article describes a community-oriented treatment program of phase-specific medical and psychosocial treatments integrated within an intensive case management model for patients with first-episode psychosis in a geographically defined population. One year status is reported for a consecutive sample of patients with nonaffective mostly schizophrenic first-episode psychosis who were receiving treatment in this program. METHODS: Patients were assessed at baseline and at one year with a modified version of the Interview for Retrospective Assessment of Onset of Schizophrenia, the Structured Clinical Assessment for DSM-IV, the Scale for Assessment of Positive Symptoms, and the Scale for Assessment of Negative Symptoms to ascertain baseline patient characteristics, remission rates, hospital readmission rates, and change in the severity of symptoms. RESULTS: Data at 13 months for 53 patients indicated a complete remission rate of 70 percent, a hospital readmission rate of 20 percent, a highly significant improvement in all dimensions of psychopathology, higher rates of remission among patients who entered treatment within six months of the onset of psychosis (82 percent compared with 60 percent), and a longer median duration of untreated psychosis among patients who did not experience complete remission (10.5 compared with 6.5 months). Nearly half the patients received initial treatment as outpatients without adverse consequences for their subsequent use of hospitalization. CONCLUSIONS: An epidemiologically representative sample of patients experiencing a first episode of psychosis, when treated optimally with low dosages of novel antipsychotics and phase-specific psychological interventions, showed a high rate of clinical recovery and were able to remain in the community most of the time. PMID- 11919361 TI - Factors influencing care seeking for a self-defined worst panic attack. AB - OBJECTIVE: Only 60 percent of persons who experience panic attacks seek treatment for them, many at the emergency department. The author documented care-seeking behaviors among persons living in the community who had experienced panic attacks and studied determinants of care seeking. METHODS: In-depth structured interviews were conducted with 97 randomly selected community-dwelling adults who met DSM III-R criteria for panic attacks. Participants were asked whether they had contemplated using or had actually used medical, alternative, and family sources of care when they had experienced their worst attack. RESULTS: Seventy-seven participants (79 percent) had considered using a general medical or mental health site when they experienced their worst attack. Of these, 50 (52 percent) had actually used such a site. General medical sites were contemplated more often (72 percent of participants) than mental health sites (27 percent), particularly emergency departments (43 percent) and family physicians' offices (34 percent). Other sources, such as friends or family members, alternative sites, and self treatment, were contemplated less often. Once contemplated, certain sources were readily used, such as ambulances, family members, and self-treatment. Several factors were significantly associated with whether a person contemplated seeking care: access or barriers to treatment, perception of symptoms and of the reasons for the panic attack, and family-related variables. CONCLUSIONS: Contemplation and use of a mental health site after a panic attack was rare among the participants in this study. Further study of determinants of care seeking may help explain why persons who experience panic attacks fail to seek treatment or seek treatment from non-mental health sources. PMID- 11919362 TI - Inpatient psychotherapy compared with usual care for patients who have schizophrenic psychoses. AB - The study examined whether some patients with schizophrenia benefit from intensive inpatient psychodynamic psychotherapy while others are harmed by it. Multiple regression analyses were conducted with combined data from a follow-up study of 25 inpatients who were treated in a psychotherapeutic program and a follow-up study of 71 patients who received standard hospital treatment. The mean duration of the follow-up period was seven years. The analyses showed that improvement in global mental health status from the index admission to follow-up was associated with the type of treatment and with the patient's clinical condition at the index admission. A strong interaction effect was found between these two variables. Psychotherapeutic inpatient programs may be beneficial to patients who have higher levels of global functioning at the start of treatment but detrimental to other patients. PMID- 11919363 TI - Factors associated with attendance at a first appointment after discharge from a psychiatric hospital. AB - A retrospective review of medical records of 158 patients who were discharged from a university-affiliated psychiatric hospital in the Midwest found that 29 patients (18 percent) did not attend their scheduled follow-up appointment. Multivariate logistic regression analysis showed that patients who were white, who lived in an urban county, and whose scheduled appointment was within two weeks of discharge were significantly more likely to attend their follow-up appointment. The study showed that patients who have an elevated risk of missing their follow-up appointment after hospital discharge can be prospectively identified. This information can be used to develop strategies for improving the likelihood that at-risk patients will receive follow-up care. PMID- 11919364 TI - A computerized patient information system in a psychiatric hospital. AB - The authors describe a computerized patient information system at a psychiatric hospital in Israel. The system is a fully implemented work instrument that promotes clinical safety and cost containment. It allows interactive online consultations, clinical cross-checking, the production of computerized reports and schedules, fast response to laboratory results, and safer drug administration, all of which help improve the quality of care. Cost savings have been achieved in areas such as pharmacy and food distribution. The initial investment in the system was $400,000, which is expected to be recouped after 11.4 years. PMID- 11919365 TI - Psychoeducational debriefings after the September 11 disaster. PMID- 11919366 TI - A second chance for people with "treatment-refractory" psychosis. PMID- 11919367 TI - The case of Sam. PMID- 11919369 TI - Firearms risk management. PMID- 11919368 TI - The case of Sam. PMID- 11919371 TI - Conditional release and mandated outpatient treatment. PMID- 11919373 TI - Risk for individuals with schizophrenia who are living in the community. PMID- 11919374 TI - Autocastration and schizophrenia. PMID- 11919376 TI - Race, ethnicity, and health: can genetics explain disparities? AB - Over the past decade, numerous studies have documented profound racial and ethnic disparities in disease in the United States. This essay examines how popular and scientific concepts of race and ethnicity converge with dominant understandings of genetics to inform the design and interpretation of research, public health policy, and medical practice. Although there is some acknowledgment in the biomedical community that racial and ethnic categories are social and not genetic, ideas about race and ethnicity that circulate in biomedicine are contradictory. Thus, in practice genetic explanations for observed differences are common both in the scientific literature and in popular media accounts of biomedical research. Such explanations naturalize racial and ethnic difference and create a conceptual barrier to developing a research program that explores the complex ways in which social inequality and experiences of racial discrimination interact with human biology to influence patterns of disease. Importantly, genetically based ideas lead to disease prevention policies that are bound to be ineffective. PMID- 11919377 TI - Perspectives on dimensional analysis in scaling studies. AB - Scaling, as defined here, refers to the precise identification of those structural and functional aspects of selected systems that are size-independent, over some specified size-range. Small and large instances of such systems are said to be similar in respect to those aspects. Physicists and engineers have developed an elaborate methodology for identifying quantitative similarity criteria applicable to physical systems. These criteria are usually derived by dimensional analysis of physical laws pertinent to a given system. Knowledge of similarity criteria allows one to predict quantitatively the behavior of a large scale prototype from measurements made on a small-scale model (e.g., in a wind tunnel). Numerous workers have sought to apply this elegant methodology to scale up in biology. After briefly reviewing dimensional analysis, scaling, and modeling, as deployed in physics and engineering, this article discusses several well-known examples of their application to bioscaling problems (chiefly in mammals) and gives reasons for doubting their validity. It concludes that this methodology is unlikely to provide explanations applicable to scale-up in diverse species. PMID- 11919378 TI - The Karamazov complex: Dostoevsky and DNR orders. AB - Families making difficult end-of-life decisions in the intensive care unit often do not exercise their autonomy in accord with the individualistic philosophic and legal models that currently prevail. Instead, they try to avoid responsibility and deny complicity, even for decisions that they ultimately approve. This paper examines two novels and a recent case from a neonatal intensive care unit that reveal how people actually make tragic decisions for family members. Dostoevsky's The Brothers Karamazov and Kenzaburo Oe's A Personal Matter explore the patterns of communication by which people in such situations test complicity and share or submerge accountability. The psychological similarities between the novelists' portrayals and the actual processes that families undergo in the ICU have practical clinical implications for the ways physicians approach discussions about do not resuscitate (DNR) orders with patients' family members. PMID- 11919379 TI - The enduring scientific contributions of Sigmund Freud. AB - Through the development of a novel observational method, Sigmund Freud made possible the collection of reliable data about man's inner life. The scientific hypotheses he formulated about these formed the initial version of psychoanalysis. Many of these first thoughts have had to be revised in the light of subsequent scientific findings about the operations of the central nervous system, but even these refuted propositions often had much heuristic value. Despite the passage of a whole century, many Freudian hypotheses have retained their scientific standing. Most important among these was Freud's realization that human thought is usually unconscious. His understanding of the role of the automatic repetition of basic patterns of behavior, of the fateful consequences of early childhood emotional vicissitudes in structuring enduring mental dispositions, and of the distinction between two distinct modes of thinking are the most significant among his many contributions. PMID- 11919380 TI - High-tech brains: a history of technology-based analogies and models of nerve and brain function. AB - This article reviews some of the technological devices and ideas which have been used over the years to answer the question, how does the brain work? It describes some of the early technology-based analogies and models of nerve fibers, and then discusses other analogies and models of the brain based on mechanical and electrical technologies. There are also short sections on cybernetics, telephone exchanges, and computers. Although all of these ideas are flawed to some extent, this article offers a brief argument on the usefulness of analogy and abstraction in brain science. PMID- 11919381 TI - Population genetics and sociobiology: conflicting views of evolution. AB - This article explores the tension between the population genetics and sociobiological approaches to the study of evolution. Whereas population geneticists, like Stanford's Marc Feldman, insist that the genetic complexities of organisms cannot be overlooked, sociobiologists (many of whom now prefer to call themselves "behavioral ecologists") rely on optimization models that are based on the simplest possible genetics.These optimization approaches have their roots in the classical result known as the fundamental theorem of natural selection, formulated by R. A. Fisher in 1930. From the start there was great uncertainty over the proper interpretation of Fisher's theorem, which became confused with Sewall Wright's immensely influential adaptive landscape concept. In the 1960s, a new generation of mathematical biologists proved that Fisher's theorem did not hold when fitness depended on more than one locus. Similar reasoning was used to attack W. D. Hamilton's inclusive fitness theory. A new theory, known as the theory of long-term evolution, attempts to reconcile the rigorous population genetics approach with the long-standing sociobiological view that natural selection acts to increase the fitness of organisms. PMID- 11919382 TI - "No practical capabilities": American biological and chemical warfare programs during the Korean war. AB - Much controversy still surrounds accusations that American forces in the Far East during the Korean War used biological warfare against North Korea and China. An analysis of recently declassified documents reveals that, although the United States attempted to accelerate its development and acquisition of such weapons during that period, its efforts to create a viable biological warfare capability were unsuccessful. Plans to similarly expand chemical warfare stocks and capabilities were also frustrated. Technological difficulties, personnel shortages, bureaucratic battles between the armed services, and policy limitations combined to hold back advances in American chemical and biological warfare. In light of the recent fears of terrorist attacks with such weapons, this analysis highlights the great difficulties involved in developing, acquiring, and delivering such capabilities. PMID- 11919383 TI - Conflicting interests in Toronto: anatomy of a controversy at the interface of academia and industry. AB - In December 2000, the University of Toronto breached a contract it held with me, initiating a sequence of events that has led to a public letter to the University from a large number of senior figures in the psychopharmacology community, protesting against the infringement of academic freedom involved, and a first ever legal action seeking redress for violation of academic freedom. This case has been intertwined from the start with a longer running debate about the possibility that the SSRI group of antidepressants may have the potential to trigger suicidality or other serious effects in a subgroup of takers. And this specific issue connects to concerns about conflict of interest in the domain of therapeutics, as well as in science in general, the ghostwriting of scientific articles, and a series of other hot-spots on the interface between academia and industry. PMID- 11919384 TI - Life or death, mad or sane--who decides? PMID- 11919385 TI - Sue's several heads: the evolution of the natural history museum. PMID- 11919386 TI - An overview of translocation-related oncogenesis in the chronic myeloid leukaemias. AB - The demonstration of the BCR-ABL fusion gene in patients with chronic granulocytic leukaemia and t(9;22)(q34;q11) represents the first recognition, in a human neoplasm, of a translocation leading to formation of an oncogenic fusion gene. Since this initial observation, this leukaemogenic mechanism has been increasingly recognized in chronic myeloid leukaemias. The fusion gene has often incorporated part of a gene encoding a receptor or cytoplasmic tyrosine kinase, particularly ABL, PDGFRB and FGFR1. This contrasts with the frequent involvement of genes encoding transcription factors or other nuclear proteins in acute myeloid leukaemia. Nevertheless, genes encoding tyrosine kinases have also been implicated in some cases of acute leukaemia. With the exception of the BCR-ABL fusion gene in chronic granulocytic leukaemia, all these fusion genes are uncommon or rare among cases of chronic myeloid leukaemia. The molecular mechanisms underlying the great majority of cases of Philadelphia-negative chronic myeloid leukaemia remain to be discovered. PMID- 11919387 TI - Cytogenetic and molecular monitoring of residual disease in chronic myeloid leukaemia. AB - For patients with chronic myeloid leukaemia, methods for monitoring response to treatment have changed considerably in recent years. In the 1980s, the principal approach was repeated examination of bone marrow metaphases for the presence of the Ph chromosome in patients treated by interferon-alpha (IFN-alpha) or allogeneic stem cell transplantation. The use of fluorescence in situ hybridisation (FISH) techniques to detect the BCR-ABL fusion gene in Ph-positive leukaemia cells increased the sensitivity of cytogenetic studies to some degree. In the last 10 years, the reverse-transcriptase polymerase chain reaction (RT PCR) has proved extremely valuable for assessing and monitoring minimal residual disease in patients who achieve Ph negativity after treatment with IFN-alpha or with the new Abl tyrosine kinase inhibitor imatinib mesylate or after allogeneic stem cell transplantation (SCT). Results are consistent with the notion that the majority of long-term survivors after allogeneic SCT are probably 'cured'; for other patients monitored serially in complete cytogenetic remission, rising numbers of BCR-ABL transcripts detected by RT-PCR can indicate the need for further therapy. PMID- 11919388 TI - Cytogenetic and molecular genetic evolution of chronic myeloid leukemia. AB - Chronic myeloid leukemia (CML) is genetically characterized by the presence of the reciprocal translocation t(9;22)(q34;q11), resulting in a BCR/ABL gene fusion on the derivative chromosome 22 called the Philadelphia (Ph) chromosome. In 2-10% of the cases, this chimeric gene is generated by variant rearrangements, involving 9q34, 22q11, and one or several other genomic regions. All chromosomes have been described as participating in these variants, but there is a marked breakpoint clustering to chromosome bands 1p36, 3p21, 5q13, 6p21, 9q22, 11q13, 12p13, 17p13, 17q21, 17q25, 19q13, 21q22, 22q12, and 22q13. Despite their genetically complex nature, available data indicate that variant rearrangements do not confer any specific phenotypic or prognostic impact as compared to CML with a standard Ph chromosome. In most instances, the t(9;22), or a variant thereof, is the sole chromosomal anomaly during the chronic phase (CP) of the disease, whereas additional genetic changes are demonstrable in 60-80% of cases in blast crisis (BC). The secondary chromosomal aberrations are clearly nonrandom, with the most common chromosomal abnormalities being +8 (34% of cases with additional changes), +Ph (30%), i(17q) (20%), +19 (13%), -Y (8% of males), +21 (7%), +17 (5%), and monosomy 7 (5%). We suggest that all these aberrations, occurring in >5% of CML with secondary changes, should be denoted major route abnormalities. Chromosome segments often involved in structural rearrangements include 1q, 3q21, 3q26, 7p, 9p, 11q23, 12p13, 13q11-14, 17p11, 17q10, 21q22, and 22q10. No clear-cut differences as regards type and prevalence of additional aberrations seem to exist between CML with standard t(9;22) and CML with variants, except for slightly lower frequencies of the most common changes in the latter group. The temporal order of the secondary changes varies, but the preferred pathway appears to start with i(17q), followed by +8 and +Ph, and then +19. Molecular genetic abnormalities preceding, or occurring during, BC include overexpression of the BCR/ABL transcript, upregulation of the EVI1 gene, increased telomerase activity, and mutations of the tumor suppressor genes RB1, TP53, and CDKN2A. The cytogenetic evolution patterns vary significantly in relation to treatment given during CP. For example, +8 is more common after busulfan than hydroxyurea therapy, and the secondary changes seen after interferon-alpha treatment or bone marrow transplantation are often unusual, seemingly random, and occasionally transient. Apart from the strong phenotypic impact of addition of acute myeloid leukemia/myelodysplasia-associated translocations and inversions, such as inv(3)(q21q26), t(3;21)(q26;q22), and t(15;17)(q22;q12-21), in CML BC, only a few significant differences between myeloid and lymphoid BC are discerned, with i(17q) and TP53 mutations being more common in myeloid BC and monosomy 7, hypodiploidy, and CDKN2A deletions being more frequent in lymphoid BC. The prognostic significance of the secondary genetic changes is not uniform, although abnormalities involving chromosome 17, e.g., i(17q), have repeatedly been shown to be ominous. However, the clinical impact of additional cytogenetic and molecular genetic aberrations is most likely modified by the treatment modalities used. PMID- 11919389 TI - Sequential transformation of t(8;13)-related disease: a case report. AB - We report a case of an atypical myeloproliferative disorder with t(8;13) that presented as B-lineage acute lymphoblastic leukaemia (B-ALL). Following induction chemotherapy, the disease manifested as chronic myeloproliferative state, which responded to hydroxyurea. Terminally, the disease transformed into acute myeloid leukaemia (AML) with additional chromosomal abnormalities including monosomy 7. To our knowledge, this is the first case of this rare atypical myeloproliferative disorder with t(8;13) that presented as B-ALL and terminally transformed to AML with additional chromosomal abnormalities. PMID- 11919390 TI - A fourth case of 8p11 myeloproliferative disorder transforming to B-lineage acute lymphoblastic leukaemia. A case report. AB - A 56-year-old male presented with inguinal lymphadenopathy and leucocytosis (WBC 98 x 10(9)/l). Bone marrow morphology showed myeloid hyperplasia, with eosinophilia. Cytogenetic analysis showed no evidence of the Philadelphia chromosome, and fluorescence in situ hybridisation studies for the BCR/ABL fusion were negative. All cells examined showed the t(8;13)(p11;q12) translocation. Six weeks after presentation, the disease progressed to an acute lymphoblastic leukaemia (ALL). The lymphoblasts were CD19/CD10 dual positive. Cytogenetic analysis again showed the t(8;13) translocation, with no additional abnormalities. There have been at least 14 reported cases of the t(8;13) myeloproliferative disorder to date, of which only 3 transformed to B-lineage ALL: our case is the 4th. PMID- 11919391 TI - The 8p11 myeloproliferative syndrome: a distinct clinical entity caused by constitutive activation of FGFR1. AB - Several recurrent translocations that involve chromosome band 8p11 have been described in myeloid malignancies. These translocations target two distinct genes: (1) FGFR1, a receptor tyrosine kinase for fibroblast growth factors, and (2) MOZ, a putative histone acetyltransferase whose precise function remains to be defined. Disruption of FGFR1 is associated with a disease entity known as the 8p11 myeloproliferative syndrome (EMS)/stem cell leukemia-lymphoma syndrome, a chronic myeloproliferative disorder that frequently presents with eosinophilia and associated T-cell lymphoblastic lymphoma. The disease is aggressive and rapidly transforms to acute leukaemia, usually of myeloid phenotype. Currently, only allogeneic stem cell transplantation appears to be effective in eradicating or suppressing the malignant clone. To date, four gene fusions associated with distinct translocations have been described in EMS: the t(8;13)(p11;q12), t(8;9)(p11;q33), t(6;8)(q27;p11) and t(8;22)(p11q22) fuse ZNF198, CEP110, FOP and BCR, respectively, to FGFR1. The resulting fusion proteins have constitutive tyrosine kinase activity and activate multiple signal transduction pathways. These pathways and the fusion proteins are attractive targets for targeted signal transduction therapy. PMID- 11919392 TI - Chronic eosinophilic leukaemia presenting with erythroderma, mild eosinophilia and hyper-IgE: clinical, immunological and cytogenetic features and therapeutic approach. A case report. AB - A 23-year-old, white male metallurgist presented with pruritic erythematous maculo-papules over the trunk and upper limbs and 6 months later developed erythroderma, eosinophilia and multi-organ dysfunction. A diagnosis of chronic eosinophilic leukaemia was made on the basis of myeloproliferative involvement of both peripheral blood and bone marrow, associated with eosinophilic differentiation and a t(5;12)(q33;p13) translocation. The initial therapeutic approach was interferon alfa-2b plus cytosine arabinoside, for 13 months, followed by hydroxyurea plus vincristine. There was improvement of skin lesions, disappearance of eosinophilia and decrease of serum immunoglobulin E, towards normal values. PMID- 11919393 TI - Myeloproliferative disorders with translocations of chromosome 5q31-35: role of the platelet-derived growth factor receptor Beta. AB - Acquired reciprocal chromosomal translocations that involve chromosome bands 5q31 33 are associated with a significant minority of patients with BCR-ABL-negative chronic myeloid leukemias. The most common abnormality is the t(5;12)(q33;p13), which fuses the ETV6/TEL gene to the platelet-derived growth factor receptor-beta (PDGFRB), a receptor tyrosine kinase that maps to 5q33. PDGFRB is disrupted by other translocations and to date four additional partner genes (H4, HIP1, CEV14 and Rab5) have been reported. Clinically, most patients present with a myeloproliferative disorder (MPD) with eosinophilia, eosinophilic leukemia or chronic myelomonocytic leukemia and thus fall into the broader category of myeloproliferative disorders/myelodysplastic syndromes (MPD/MDS). With the advent of targeted signal transduction therapy, patients with rearrangement of PDGFRB might be better classified as a distinct subgroup of MPD/MDS. PMID- 11919394 TI - Cytogenetic and molecular genetic abnormalities in systemic mastocytosis. AB - Clonal cytogenetic abnormalities similar to those observed in other haematological neoplasms are demonstrable in a proportion of patients with systemic mastocytosis and in a smaller proportion of adults with urticaria pigmentosa without apparent systemic disease. These clonal abnormalities are not likely to represent the primary event in mast cell neoplasms. Although no recurrent cytogenetic abnormalities specific for mastocytosis have yet been recognized, the majority of cases display one of several mutations in c-KIT, the gene encoding the receptor for stem cell factor. That most commonly observed is the Asp816Val mutation, which permits proliferation independent of growth factors. c-KIT mutations may play a significant role in the biology of mast cell malignancies, although other mutations may be needed for a malignant phenotype. PMID- 11919395 TI - Efficacious response with low-dose indapamide therapy in the treatment of type II diabetic patients with normal renal function or moderate renal insufficiency and moderate hypertension. AB - We examined the efficacy of low daily dose (1.25 mg/day) of indapamide in the treatment of high blood pressure in patients with diabetes mellitus with normal renal function and those with moderate renal insufficiency (serum creatinine <1.5 mg/dl). The study was an open label one of four months duration. Twenty-eight patients were enrolled in the study and only 15 completed it. Within 2 weeks of therapy, systolic blood pressure fell from 173 +/- 4.5 to 144 +/- 2.0 mm Hg and diastolic blood pressure from 96 +/- 2.1 to 80 +/- 1.8 mm Hg (p < 0.01) and blood pressure remained at these levels throughout the study. The results show that low dose indapamide is effective in the treatment of moderate hypertension in patients with diabetes mellitus who have normal renal function or moderate renal insufficiency. Therefore, this dose of 1.25 mg/day is recommended for the treatment of such patients. PMID- 11919396 TI - Evaluation of superoxide dismutase activity in dialyzed patients by electron spin resonance spectroscopy. AB - BACKGROUND: The relationship between plasma SOD activity and remaining renal function in patients with renal insufficiency has been not elucidated. We attempted to investigate the degree of serum SOD activity among patients with various renal conditions. METHODS: Using electron spin resonance (ESR) spectroscopy, plasma SOD activities were evaluated in 28 patients who underwent HD before and after procedure and in 15 controls. The patients were classified according to the condition of renal function; group A (n = 15) included patients whose plasma creatinine clearance levels were less than 10 ml/min, and group B (n = 13) included those whose plasma creatinine clearance levels were 10 ml/min or more but less than 20 ml/min. RESULTS: The plasma SOD activities of group A were significantly higher than those of group B (p < 0.005) and controls (p < 0.001). Multivariate analysis showed that in group A, the SOD activities in plasma before HD were significantly higher than those after HD (p < 0.001). However, in group B, there were no significant differences in the level of plasma SOD activities before and after HD. CONCLUSION: Production of superoxide anion is considered almost the similar in both groups, which suggests the SOD activity would depend on the degree of the remaining renal function of each patient. PMID- 11919397 TI - Substance use among Iranian nephrologic patients. AB - AIMS: This study assessed the prevalence of substance use among Iranian patients with nephrologic disease (chronic renal failure) who were admitted in different nephrologic wards at Shiraz general hospitals. DESIGN: Cross-sectional survey using structured interview and also using DSM-IV criteria for substance dependency. SETTING: General hospitals in Shiraz city (patients with nephrologic disease admitted in different nephrologic wards). PARTICIPANTS: 64 (32 men and 32 women) patients selected randomly. FINDINGS: Data were gathered by a structured interview from 64 patients admitted in nephrologic wards of general hospitals in 2001. The mean age was 49.39 years (SD = 19.62) ranging from 18 to 80 years. 26 (40.6%) of the subjects (65.6% of the men and 15.6% of the women) reported the use of substance(s) once or more in their lives. The majority, 23 (35.9%) used tobacco, 9 (14.1%) used opium and 2 (3.1%) used alcohol. None had used cannabis, heroin, cocaine or LSD. 20 (31.3%) of the subjects (50% of the men and 12.5% of the women) were currently substance dependent, using DSM-IV criteria. The majority, 18 (28.1%) were nicotine dependent, and 6 (9.4%) were opium dependent. There was a nonsignificant relationship between income or occupation or education and prevalence of substance use. The reported reasons for initial use of substance(s), in order of frequency, were enjoyment, modeling and release of tension, and also for current users were, habit, enjoyment and need. CONCLUSIONS: Substance use especially cigarettes, opium and alcohol was found to be high among patients. There was no report of cannabis, heroin, cocaine or LSD use. Cultural attitudes toward substance use were found to affect the type and amount of use. These findings can be considered when planning preventive or therapeutic programs. PMID- 11919398 TI - Bone mineral density in patients beginning hemodialysis treatment. AB - BACKGROUND: Our study was designed to determine bone mineral density (BMD) in patients beginning hemodialysis (HD) treatment, a possible correlation with the duration of renal failure prior to treatment, a possible correlation with the basic disease and the association with the concentration of intact parathormone (iPTH). METHODS: Our prospective clinical trial included 50 patients beginning HD treatment. Cortical bone mineral density (BMDc) was measured at the left femoral neck and trabecular bone mineral density (BMDt) in the region of the lumbosacral spine. Bone mineral density (BMD) was measured by quantitative digital radiography using a Hologic 2000 plus device belonging to the third generation of densitometers based on dual-energy X-ray absorptiometry. RESULTS: In patients (PTS) beginning HD, the average BMDc was 82 +/- 15% of BMDc in a healthy population of corresponding age and sex. The average BMDt was 91 +/- 16% of BMDt in a healthy population of corresponding age and sex. The difference was statistically significant (p < 0.05). There is a negative correlation between iPTH and BMDc r = -0.34 (p < 0.02). Patients with chronic glomerulonephritis (GN) had a statistically significantly higher BMDc (g/cm(2)) (p < 0.01) than those with analgetic nephropathy (AN). PTS with AN have lower BMDc (g/cm(2), %) (p < 0.02) and BMDt (p < 0.005) than the rest of the PTS, iPTH in PTS with AN is higher than in the rest of the PTS (p < 0.05). CONCLUSIONS: In PTS at the beginning of HD, BMD is lower than in healthy people of corresponding age and sex. This means that BMD already decreases prior to HD. BMDc was statistically significantly lower than BMDt (p < 0.00005). PTS with AN have lower BMD than those with GN and all remaining PTS. A negative correlation between iPTH and BMDc was found. PMID- 11919399 TI - Correlates of sleep behavior among hemodialysis patients. The kidney outcomes prediction and evaluation (KOPE) study. AB - BACKGROUND: Given the importance of sleep to overall physical and mental well being, we sought to identify the correlates of seven sleep disturbances in a cross-sectional study of a biracial population of male and female patients treated with hemodialysis. METHODS: Univariate and multivariate analyses of a cohort study of adult patients with end-stage renal disease. Demographic, psychosocial, clinical, and health and physical functioning variables were included. RESULTS: Waking up during the night (57%) and waking up too early (55%) were the most commonly reported sleep problems. Multivariate analyses consistently indicated that levels of pain, depressive symptoms, and physical functioning were consistently associated with the seven sleep disturbances. CONCLUSION: Sleep disturbances are common in patients with end-stage renal disease. Physical and mental well-being were consistently related to the seven sleep disturbances. PMID- 11919400 TI - Current Canadian approaches to dialysis for acute renal failure in the ICU. AB - BACKGROUND: Although there is a very high mortality rate (>50%) with acute renal failure (ARF) in the intensive care unit (ICU), there is no general consensus on the best dialysis treatment for this condition. METHODS: We surveyed by mail questionnaire, all adult academic and community registered Canadian nephrology centers that offer treatment for ARF. RESULTS: The overall response rate was 59% (53/90). Comparing current dialysis methods with those utilized 5 years ago, the largest increase was in continuous renal replacement therapies (CRRT) (26 vs. 9%). Both intermittent hemodialysis (IHD) and peritoneal dialysis decreased in utilization. The predominant current CRRT methods utilized venovenous access (80%), as compared to 5 years ago when arteriovenous was the most common (52%). Despite data from chronic dialysis (and preliminary data in ARF) suggesting reduced mortality and morbidity with increasing dialysis dose, there was no formal method of dialysis prescription monitoring in over 75% of the centers. CONCLUSION: Notwithstanding a lack of definitive evidence of superior outcomes with CRRT compared to older methods, the utilization of CRRT is dramatically increasing for the treatment of ARF in Canada. Whether this shift towards CRRT, and whether more attention to dialysis dose in ARF, might be expected to lead to better outcomes, requires further evaluation. PMID- 11919401 TI - Prevalence of nephropathy in black patients with type 2 diabetes mellitus. AB - The prevalence of nephropathy in black patients with type 2 diabetes mellitus is poorly defined. We performed a cross-sectional analysis of 98 unrelated and unselected black type 2 diabetic patients treated in indigent care internal medicine clinics to determine the prevalence of proteinuria and nephropathy. Serum creatinine, blood urea nitrogen, urine albumin and urine creatinine concentrations were measured. A Spearman's rank correlation was computed to test for a relationship between diabetes duration and continuous outcomes. For binary outcomes, an odds ratio and 95% confidence interval were computed for a change of 10 years diabetes duration based on logistic regression. Cases were 61% female, and had mean (+/- SD) age 59.9 +/- 12.5 years, diabetes duration 12.6 +/- 9.4 years, body mass index 32.4 +/- 9.3 kg/m(2), hemoglobin A1C (HbA1C) 9.2 +/- 2.3%, and serum creatinine concentration 1.60 +/- 1.1 mg/dl. For continuous variables, diabetes duration was positively associated with albuminuria (r = 0.31; p = 0.0017), serum creatinine (r = 0.36; p = 0.0003) and blood urea nitrogen concentration (r = 0.36; p = 0.0003). For binary variables, cases with longer diabetes duration were at increased risk for urinary albumin:creatinine >300 microg/mg (p = 0.006), elevated serum creatinine concentration (> or = 1.4 mg/dl in women or > or = 1.6 mg/dl in men; p = 0.045), elevated blood urea nitrogen concentration (> or = 20 mg/dl; p = 0.026), and clinical cerebrovascular disease (p = 0.028). HbA1C, body mass index, and blood pressure did not correlate with diabetes duration in this population. Among the cases, 33.7% had elevated serum creatinine concentration and 71.5% had abnormal levels of albuminuria (27.6% > 300 microg albumin/mg Cr and 43.9% 30-300 microg albumin/mg Cr). Abnormal proteinuria was seen in the majority of black patients with poorly controlled type 2 diabetes mellitus treated in indigent care clinics. This prevalence may be conservative, due to the widespread use of angiotensin-converting enzyme inhibitor therapy and exclusion of cases treated only by nephrologists. Approximately 70% of black patients with type 2 diabetes cared for in indigent care clinics have abnormal proteinuria and are at heightened risk for ESRD and death. PMID- 11919402 TI - Correlation between soluble markers of endothelial dysfunction in patients with renal failure. AB - AIM: Damage to the endothelium is an important component of atherosclerosis. It has been suggested to be quantified by measuring plasma markers, such as von Willebrand factor and thrombomodulin and soluble adhesion molecules. We hypothesized there may exist a correlation between the plasma levels of von Willebrand factor and thrombomodulin as markers of endothelial cell dysfunction and the serum concentrations of soluble adhesion molecules and monocyte chemoattractant protein-1 (MCP-1) in patients with renal insufficiency, and in patients on peritoneal dialysis or hemodialysis since these three groups of kidney patients are highly prone to develop cardiovascular diseases. RESULTS: The concentrations of von Willebrand factor and thrombomodulin in plasma were significantly higher in patients with kidney diseases as compared to healthy subjects (p = 0.017 and p < 0.001, respectively). The patients also had significantly higher concentrations of soluble vascular cell adhesion molecule-1 (sVCAM-1) and MCP-1 compared to healthy controls (p < 0.001 for both comparisons). There were strong correlations between the concentration of soluble intercellular adhesion molecule-1 (sICAM-1) and von Willebrand factor in patients with kidney failure (r = 0.63, p < 0.001) and between the concentration of thrombomodulin and sVCAM-1 (r = 0.61, p < 0.001). Furthermore, a negative correlation was observed between the concentration of thrombomodulin and the cell surface expression of CD11b on monocytes and granulocytes in the peripheral circulation (p < 0.01 in both cases). CONCLUSION: The strong correlation between markers of endothelial dysfunction and soluble adhesion molecules in patients with renal insufficiency and on dialysis strengthen the view that an ongoing stress on endothelial cells is present in this group of patients. This may play a pathophysiological role in the development of cardiovascular disease. PMID- 11919403 TI - High endothelial venule-like vessels in the interstitial lesions of human glomerulonephritis. AB - To clarify the characteristics of high endothelial venule (HEV) -like vessels in the interstitium of human glomerulonephritis, we investigated the expression of HEVs related molecules such as P-selectin and L-selectin ligands; MECA-79 epitope and variant sulfated forms of sialyl Lewis X (variant sLe(X), clones 2H5, 2F3, GS 13 and GS-36) in kidney specimens by means of immunohistochemical studies, and P selectin and hevin mRNA signals by using in situ hybridization analyses. In lymphoid organs, HEVs strongly expressed P-selectin, MECA-79, variant sLe(X) and hevin mRNA signals. In normal kidneys (n = 4), only P-selectin was faintly positive in the vessels of interstitium, but other molecules could not be detected. Interstitial P-selectin expression was upregulated in patients with tubulointerstitial diseases (n = 4) and proliferative glomerulonephritis (n = 51) such as IgA-related nephropathy (n = 39), membranoproliferative glomerulonephritis (n = 4) and crescentic glomerulonephritis (n = 2), but not in nonproliferative glomerular diseases (n = 39) such as minimal change nephrotic syndrome (n = 18) (1.00 +/- 0.41, 0.64 +/- 0.11, 0.21 +/- 0.05, respectively). Interstitial P-selectin expression also correlated with interstitial local cellular infiltration (r = 0.60, p < 0.0001). In addition, P-selectin mRNA signals were detected on the peritubular capillaries and HEV-like vascular endothelial cells. MECA-79 and variant sLe(X) (2H5 and 2F3) were weakly expressed on the HEV-like vessels located at the corticomedullary regions in three cases (7%) and in nine cases (27%) with interstitial cellular infiltration, respectively. However, we could not detect GS-13, GS-36 or hevin mRNA signals in the diseased kidney specimens. In conclusion, HEV-like vessels in renal interstitium expressed molecules somewhat different from HEVs in lymphoid organs and were associated with interstitial leukocyte accumulation in human proliferative glomerulonephritis possibly through the de novo expression of P selectin and partly L-selectin ligands (MECA-79 epitope and variant sLe(X)) in the interstitial lesions. PMID- 11919405 TI - Continuous intravenous sodium ferric gluconate improves efficacy in the maintenance phase of EPOrHu administration in hemodialysis patients. AB - Although intravenous iron has proved to optimize the efficacy of EPOrHu in hemodialysis patients, hitherto no consensus exists with respect to the best regimen of intravenous iron administration. We started a prospective randomized study in 26 patients undergoing chronic hemodialysis who had adequate iron metabolism indices (serum ferritin >100 microg/l; %TSAT >20%; %HypoE <10% and CHr >26 pg) and were in the maintenance phase of EPOrHu administration (target hemoglobin obtained >10 g/dl). All patients were receiving sodium ferric gluconate (Ferrlecit) intermittently prior to the study and after a 1-month wash out period where iron was not administered patients were randomized to receive the same previous dose of intravenous iron either in a continuous (6.25-21.3 mg in every hemodialysis session) or an intermittent regimen (62.5 mg every 1-4 weeks, not modifying the previous schedule of administration). At 16 weeks, the continuous group showed a significant increment in serum Hb (11.83 +/- 1.12 g/dl) with respect to baseline (10.96 +/- 1.31 g/dl) (p < 0.05), whereas no differences were obtained in intermittent group (baseline: 11.16 +/- 1.03 g/dl; 16 weeks: 11.14 +/- 0.90 g/dl, NS). In contrast with the intermittent group, serum ferritin increased significantly in the continuous group (16 weeks: 508 +/- 157 microg/l; baseline: 368 +/- 56 microg/l; p < 0.05), whereas %TSAT and CHr did not modified during the study in both groups. %HypoE increased significantly with respect to baseline values in the continuous group (p < 0.05) and close to significantly different in the intermittent group (p = 0.06). Our study suggests that hemodialysis patients in the maintenance phase of EPOrHu administration would obtain further benefit in terms of serum hemoglobin level with a continuous intravenous serum ferric gluconate regimen, at least in the short term. PMID- 11919404 TI - Comparison between tinzaparin and standard heparin for chronic hemodialysis in a Canadian center. AB - BACKGROUND: Low-molecular-weight heparins offer several advantages over standard heparins, but their use for maintenance hemodialysis has been limited in North America because of their higher cost. Our objective was to compare tinzaparin to standard heparin during maintenance hemodialysis over an 8-week period, in regard to the visual aspect of the extracorporeal circuit, filter reuse, bleeding and time for compression of vascular access at the end of hemodialysis session, nursing time devoted to anticoagulation administration, level of satisfaction of patients and nurses, and relative cost. METHODS: Thirty-two chronic hemodialysis adult patients with peripheral accesses were randomly divided into two groups in a cross-over design: tinzaparin for 4 weeks followed by standard heparin for 4 weeks, or vice versa. Hemodialysis was performed thrice weekly over 3.5-4 h using large surface reused filters. Standard heparin was administered as an initial bolus of 50-75 units per kilogram followed by an infusion to maintain an activated clotting time (ACTESTER) between 150 and 200 s and discontinued 30-45 min before the end of the session. The initial dose of tinzaparin was 3,500 IU anti-Xa for patients usually receiving 7,500 units or less of standard heparin, or 4,500 IU anti-Xa for patients receiving more than 7,500 units of standard heparin, and it was injected as a bolus in the arterial line at the beginning of hemodialysis. Dosage adjustments were made by increments or decrements of 500 IU. RESULTS: A total of 6 patients did not require any adjustment in their dose of tinzaparin and remained at the initial dose, while the remaining 26 necessitated adjustments of the initial dose of tinzaparin: 20 patients required increments from the initial dose whereas 6 required reductions. For most patients, 27 of them, the standard heparin dose was kept at the same level throughout the study period (since it was their usual regimen and they were in stable medical conditions). According to the monitoring scale, the visual aspects of the tubing of the extracorporeal circuit and of the dialyzers at the end of the session were similar for both tinzaparin and standard heparin. The time of compression of the vascular access at the end of the hemodialysis sessions was not significantly different with tinzaparin than with standard heparin. However, as indicated below, most patients noted less bleeding (or oozing) from their access (during compression and thereafter, in the few hours after hemodialysis) with tinzaparin than with standard heparin. Clotting was observed more frequently in the arterial and venous bubble traps with tinzaparin than with standard heparin. The presence of clot(s) was observed in the arterial and venous bubble traps in, respectively, 18 +/- 12 and 10 +/- 6% of the sessions with tinzaparin, while in, respectively, 3 +/- 4 and 2 +/- 4% of the sessions with standard heparin (p < 0.005). Despite a tendency for a reduced reuse number of the dialyzers, the difference did not reach statistical significance. Among the 30 patients who completed the study, 2 reported excessive bleeding from their vascular access with tinzaparin whereas 8 reported such an excessive bleeding with standard heparin. The level of satisfaction of patients and nurses for tinzaparin was extremely good. The main reasons stated by the patients was reduced bleeding from their access after dialysis. The nurses preferred tinzaparin because of the simplicity and the rapidity of its administration, the lack of monitoring required, and the decreased bleeding/oozing tendency from the vascular access sites. The time spent for anticoagulation during a hemodialysis session was reported as 5 min with standard heparin (if no ACTESTER monitoring), 25-30 min with standard heparin (if ACTESTER monitoring required), and 1 min with tinzaparin. The cost analysis revealed that although tinzaparin is more than six times more expensive than standard heparin, the use of tinzaparin becomes similar to the use of standard heparin (USD 7.33 vs. USD 7.62 Canadian dollars for one hemodialysis session) if ACTESTER monitoring is performed (assuming that 22% of the sessions are routinely monitored and that one ACTESTER device is necessary for 8-10 dialysis stations, as applied in our unit). CONCLUSION: Our experience with tinzaparin was positive: it represents a simple and easy way to offer anticoagulation during maintenance hemodialysis, it seems associated with less postdialysis bleeding, it saves precious nursing time and is widely appreciated by patients and staff. PMID- 11919407 TI - Severe rhabdomyolysis due to malignant hyperthermia during renal transplantation procedure can cause delayed graft function. AB - A case of rhabdomyolysis from malignant hyperthermia occurred during renal transplantation surgery is presented. After the completion of vascular and uretherovesical anostomosis, the patient's heart rate began to rise, sweatiness was observed and body temperature increased to 41 degrees C. Additionally, metabolic and respiratory acidosis and hyperkalemia were detected. Serum creatine kinase and lactic dehydrogenase levels were increased significantly. After external cooling and the administration of dantrolene sodium, body temperature and heart rate were decreased. During this period; furosemide, mannitol and sodium bicarbonate were given. Three hours after the completion of surgery, urine output was begun and urine myoglobin was found to be positive. Renal function improved gradually and serum creatinine level decreased to 1.6 mg/dl on the 14th postoperative day. Malignant hyperthermia can lead to severe rhabdomyolysis and delayed graft function in renal transplant recipients. Early diagnosis and intervention is crucial for protecting renal function. PMID- 11919406 TI - Role of soluble receptors for tumor necrosis factor alpha in the development of hypoalbuminemia in hemodialysis patients. AB - BACKGROUND: Hypoalbuminemia is a significant predictor of mortality in patients on hemodialysis (HD). The cause of hypoalbuminemia in HD patients, however, remains to be clarified. Recent studies have demonstrated that high blood concentrations of soluble receptors for tumor necrosis factor (sTNFRs) are associated with malnutrition in a variety of diseases and that the blood sTNFRs concentrations are elevated in HD patients. METHODS: The serum concentrations of tumor necrosis factor alpha, sTNFR (p55 and p80), and interleukin (IL) 6 were measured in 21 HD patients with low (equal to or less than 3.6 g/dl) and in 19 HD patients with normal (equal to or more than 4.0 g/dl) concentrations of serum albumin who were free from acute infection, malignancy, collagen diseases, liver diseases, or surgery. The correlation between these parameters and the degree of hypoalbuminemia was examined. RESULTS: The serum concentrations of sTNFR p80 and IL-6 were significantly higher in patients with hypoalbuminemia as compared with those with normoalbuminemia (sTNFR p80: 47.4 +/- 4.7 vs. 35.3 +/- 2.1 ng/ml, p < 0.05; IL-6: 10.8 +/- 2.0 vs. 6.3 +/- 0.5 pg/ml, p < 0.05). In contrast, there was no difference in the serum concentrations of tumor necrosis factor alpha and sTNFR p55 between the two groups. Multivariate regression analysis showed that sTNFR p80 but not IL-6 significantly influenced the serum albumin concentrations. There were no significant differences in body mass index, serum total cholesterol, and normalized protein catabolic rate between the two groups. CONCLUSIONS: Our results suggest the development of hypoalbuminemia in some HD patients who do not have any obvious cause of hypoalbuminemia and that high concentrations of sTNFR p80 might contribute to the development of hypoalbuminemia in patients on long-term HD. PMID- 11919408 TI - Effects of aortic stenosis on renal renin, angiotensin receptor, endothelin and NOS gene expression in rats. AB - BACKGROUND/AIMS: Published data regarding the effects of common cardiovascular diseases, i.e. aortic stenosis on renal regulation of major vasoconstrictive (renin, endothelins) and vasodilatory systems (NO) are controversial. Therefore we aimed to evaluate the effects of chronic aortic stenosis on the renal renin angiotensin, endothelin and NO systems. METHODS: Experimental supravalvular aortic stenosis was induced by using silver clips with a 0.6 mm internal diameter on the ascending aorta of weanling rats. Renal endothelin-1 (ET-1), endothelin-3 (ET-3), renin, AT(1a), AT(1b), eNOS, and bNOS gene expression were assessed by RNase protection assay. RESULTS: Renal renin gene expression increased twofold in rats with aortic stenosis. In contrast, renal ET-1, ET-3, eNOS, bNOS, and AT(1a), AT(1b) gene expression were unchanged in rats with aortic stenosis. CONCLUSION: Our study demonstrates that in rats with severe experimental supravalvular aortic stenosis only renal renin gene expression is stimulated. This contrasts with severe heart failure where endothelins and NO synthases are also upregulated. Different patterns of regulation of renal vasoactive mediators may be of importance for the extent of the renal impairment associated with aortic stenosis, and may be correlated with the severity of congestive heart failure. PMID- 11919409 TI - Migration, matrix production and lamellar bone formation of human osteoblast-like cells in porous titanium implants. AB - The goal of this study was to characterize growth, mineralization and bone formation of osteoblast-like cells in titanium pore channels of defined diameter. Titanium implants with continuous drill channels of diameters of 300, 400, 500, 600 and 1,000 microm were inserted into human osteoblast-like cell cultures. The ingrowth of the cells into the drill channels was investigated by transmitted light microscopy and scanning electron microscopy. Immunofluorescence and histological analysis of 15-channel sections of each diameter were used to investigate the growth behavior and the matrix protein patterns. Mineralization was evidenced by Alizarin red staining and high-resolution microradiography. The ingrowth of human osteoblast-like cells in the drill channels occurred in a sequence of four characteristic stages. In stage 1, osteoblast precursor cells adhered to the wall of the channel and migrated three-dimensionally into the channel by forming foot-like protoplasmic processes. For all 15 sample drill channels that were investigated, the cell ingrowth over 20 days amounted on average to 793 microm (+/- 179) into 600-microm-diameter channels, where they migrated significantly faster than in all the other channels. In stage 2, approximately on day 5-7, the osteoblast-like cells began to anchor on the substrate wall by matrix proteins and to build up a dense network of matrix proteins in the drill channel. The mineralization of the extracellular matrix, while depending on cell stimulation, was initiated in stage 3, on average after 4 weeks. In drill channels of a diameter of 1,000 microm the cell growth was incomplete and no mineralization was found by radiological assessment. Starting in week 6, in the drill channels of diameters ranging from 300 to 600 microm, the network of extracellular matrix proteins and osteoblast-like cells began to form an osteon-like structure. Neither the highly developed migration behavior of osteoblastic cells nor the reorganization from a fiber-like matrix to a lamellar structure have so far been described for cell cultures. PMID- 11919410 TI - Comparative evaluation of nuclear morphology of equine oocytes aspirated in vivo and stained with Hoechst and orcein. AB - Nuclear maturation of equine oocytes was assessed immediately after in vivo collection. A double-staining technique (Hoechst and orcein) was used on the same oocytes to visualize nuclear morphology, i.e. to evaluate the chromatin configurations of each oocyte after Hoechst in relation to the nuclear morphology after orcein staining. The proportion of oocytes evaluated as germinal vesicle stages was significantly (p < 0.02) lower after Hoechst (14.5%) than after orcein staining (29.0%), while the incidence of the so-called dense chromatin stage was assessed to be higher (p < 0.05) after Hoechst than after orcein staining (14.5 vs. 6.5%). There was no difference between Hoechst and orcein staining in the incidence of diakinesis and germinal vesicle breakdown stages, respectively (44.9 vs. 42.0%), and the same applied for metaphase I (11.6 vs. 8.0%), metaphase II (7.2 vs. 8.0%) and degenerated stages (7.2 vs. 6.5%). It was concluded that the interpretation of the meiotic stages may differ between Hoechst and orcein staining and in a large proportion of equine oocytes the nuclear border may not be visualized on orcein staining. PMID- 11919411 TI - Employment of confocal microscopy for the dynamic visualization of domes in intact epithelial cell cultures. AB - Many epithelial cells cultured on plastic ware form domes, fluid-filled localized raisings of the cell monolayer. Domes are due to active vectorial ion transport and their presence demonstrates the maintenance of a differentiated polarized phenotype and of tight junctional complexes. Through a confocal laser microscope equipped with a special flow chamber, intact domes were evaluated in real time for prolonged experimental periods. Both in CAPAN-1 pancreatic duct adenocarcinoma cells and in renal tubular LLC-PK1 cells, vertical sections of calcein-loaded cultures provided a clear visualization of dome outlines during the slow deflation induced by specific agonists (respectively, 1 microM secretin or 10 microM vasopressin). Section series of calcein-loaded domes were used for three-dimensional reconstructions. In CAPAN-1 cultures, cell depolarization induced by secretin was detected with the potentiometric dye bis-oxonol. In the same cells pyranine, a fluid phase marker that is cell impermeant, visualized dome compartment and paracellular pathways, also providing an evaluation of dome fluid pH. Confocal laser scanning microscopy of domes represents a convenient device for the functional assessment of living epithelial cells. PMID- 11919412 TI - Adenosine triphosphatase expression correlates with ciliary activity of respiratory epithelium in vitro. AB - In vitro culture of respiratory epithelium is of great utility for pharmacological investigations and tissue engineering. Up to now, the degree of differentiation of respiratory cells cultured in vitro has exclusively been estimated by measuring ciliary beat frequency (CBF). Ciliary motility is dependent on the function of the motor protein dynein that is composed of at least two heavy chains, sharing attributes of adenosine triphosphatases (ATPases). CBF is further dependent on medium conditions and does not allow to draw any accurate conclusion on the proportion of fully differentiated ciliated cells in culture. For this reason we introduced the immunohistochemical detection of a 100-kD ATPase subunit as a correlation with dynein activity in human respiratory cell tissue culture. Our results show that the amount of immunohistochemically detectable ATPase-subunit-positive cells strongly correlates with ciliary motility in vitro. Cultures without ciliary activity exhibited no ATPase staining, whereas in cell cultures with excessive ciliary beat, up to 15.1% of the cells were ATPase positive. Immunohistochemical detection of ATPase in respiratory cell cultures seems to be a sensitive and reproducible complement for the characterization of cultured ciliated epithelium. PMID- 11919413 TI - Electron microscopy of the canine corneal basement membranes. AB - The purpose of this study was to characterize the surface topographical features of the epithelial and endothelial (Descemet's) basement membranes of the canine cornea. Corneas were obtained from young, healthy dogs (<2 years old) with no history or evidence of previous ocular disease. The epithelium and endothelium was carefully removed preserving the anterior and posterior basement membranes. The specimens were examined by transmission electron microscopy and scanning electron microscopy. The epithelial and endothelial basement membrane surface topography is an intricate meshwork of pores and fibers measuring in the nanometer size range. The features of the endothelial basement membrane overall are smaller in size than the epithelial basement membrane. These surface topographical features may incite changes in epithelial and endothelial cell behavior. PMID- 11919414 TI - Immunocytochemical light- and electron-microscopic studies of growth hormone, prolactin and somatomammotroph cells in female goat. AB - We have studied the structural and ultrastructural changes that occur in growth hormone (GH), prolactin (PRL) and somatomammotroph (SMT) cells of female goats of the Murciano-Granadina breed during different physiological stages: prepuberty, anestrus, pregnancy and lactation. Serial sections were stained with double immunolabeling, alkaline phosphatase and avidin-biotin complex for light microscopy, and immunogold labeling was used for electron microscopy. These techniques allowed the identification of GH, PRL and SMT cells, which were evenly distributed throughout the whole pars distalis. PRL cells were the most frequent. Both PRL and GH cells showed morphological changes that may be related to a given physiological stage. These changes include the number of cells and the number and size of the secretory granules. SMT cells were clearly identified by having two types of monohormonal secretory granule showing single labeling, in spite of their similar ultrastructural characteristics to the other adenohypophysary cells. They are found in low percentages (0.6%). We postulate that cell aggregates which look like SMT syncytia may be artifacts caused by interdigitation of PRL and GH cells, and that cells which are normally taken to be SMT cells are not an interconversion stage between monohormonal cells. PMID- 11919415 TI - Age-related morphological changes in squamous and parietomastoid sutures of human cranium. AB - Age-related morphological changes in the inner and outer surfaces of the squamous and parietomastoid sutures were examined in 65 skulls (35 male, 30 female) obtained from Japanese subjects 5-90 years of age at the time of death. Dimensions were measured in both the horizontal and sagittal planes. Wavelength analysis was done by three-dimensional construction. Irregularities were evaluated by calculating fractal dimensions. The outer squamous suture showed no significant age-related changes in size, but wavelength increased because of the development of bony interdigitations with aging. The posterior part of the outer squamous suture showed significant age-related increases in both wavelength (p < 0.01, R(2) = 0.164) and fractal dimension (p < 0.01, R(2) = 0.101) in males. The parietomastoid suture showed significant age-related changes in size (p < 0.05), but not in wavelength or fractal dimension. These distinct morphological changes found on both the inner and outer surfaces of the squamous and parietomastoid sutures suggest that the age-related morphological characteristics of these sutures are affected by extrinsic mechanical forces. PMID- 11919416 TI - Ultrastructure of human mast cells. AB - Ultrastructural studies of human mast cells define their organelles and the impact of environment, development and function on their ultrastructural morphology. The studies we review here implicate lipid bodies in eicosanoid and cytokine biology, and extend the functional repertoire of secretory-storage granules to synthesis. PMID- 11919417 TI - Regulation of mast cell phenotype by MITF. AB - The development of mast cells is controlled through the cooperative effects of growth factors and nuclear transcription factors. The signals generated by the binding of stem cell factor (SCF) to c-kit receptor tyrosine kinase (KIT) are essential for their development and survival. A double gene dose of mutant alleles at either the SCF or KIT locus results in a decrease of mast cells. A double gene dose of mutant alleles at the mi transcription factor (MITF) locus also results in mast cell deficiency. Although the phenotype of the few mast cells remaining in SCF and KIT mutant mice appeared to be normal, the phenotype of mast cells was abnormal in MITF mutant mice. We describe here the abnormalities of mast cells observed in MITF mutant mice. PMID- 11919418 TI - Kit and c-kit mutations in mastocytosis: a short overview with special reference to novel molecular and diagnostic concepts. AB - Mastocytosis is a heterogeneous group of hematopoietic disorders characterized by abnormal growth and accumulation of mast cells (MC) in one or more organs. Clinical symptoms occur as a result of the release of chemical mediators and/or of pathologic infiltration of MC in various tissues. Although the initial events leading to mastocytosis have not yet been unraveled, acquired alterations in the c-kit gene coding for the receptor of stem cell factor (SCF), a major cytokine involved in MC growth, have been described in a significant number of patients. Of particular interest are point mutations resulting in a constitutively activated SCF receptor. Such mutations are probably involved in the abnormal (SCF independent) proliferation of MC in these patients. New therapeutic strategies may be envisaged to inhibit the deregulated kinase activity of these mutant forms of c-kit. PMID- 11919420 TI - Mast cell hyperplasia: role of cytokines. AB - Mast cell hyperplasia is found in different pathologies such as chronic inflammatory processes, fibrotic disorders, wound healing or neoplastic tissue transformation. The functional significance of the accumulation of mast cells in these processes is largely unknown. It is now established that bone marrow derived mast cell progenitors circulate in peripheral blood and subsequently migrate into the tissue where they undergo final maturation under the influence of local microenvironmental factors. Cytokines are of particular importance for mast cell recruitment, development, and function. Stem cell factor (SCF) is a unique mast cell growth factor, since mast cells disappear completely in the absence of SCF. However, several other cytokines such as IL-3 and IL-4 have been shown to influence mast cell proliferation and function also. This review focuses on the role of cytokines in the regulation of mast cell hyperplasia. PMID- 11919419 TI - Histopathological and immunohistochemical aspects of mastocytosis. AB - The diagnosis of mastocytosis is based on histological evidence of a focal increase in tissue mast cells. Immunohistochemical staining with antitryptase antibodies is strongly recommended in all cases of suspected mastocytosis because mast cell infiltrates may be small and scanty. Mastocytosis may be difficult to distinguish from other hematological malignancies, in which an increase in mast cells is frequently seen. The expression of the T cell-associated antigen CD2 has been shown to be exclusively found on neoplastic mast cells in mastocytosis. The demonstration of expression of vascular endothelial growth factor by mast cells is consistent with the finding of angiogenesis which is commonly seen in tissue infiltrates of mastocytosis. PMID- 11919421 TI - Mastocytosis and fibrosis: role of cytokines. AB - Mastocytosis is a rare stem cell disorder characterized by abnormal growth and accumulation of mast cells in one or more organ systems. Clinical heterogeneity is a hallmark of mastocytosis. Recent observations of activating mutations in c kit may help to understand the abnormal growth of mast cells in mastocytosis. However, this mutation alone does not explain the entire clinical heterogeneity of the disease. Reticulin fibrosis is also commonly associated with systemic mastocytosis. Mast cells are known to be the source of fibrogenic cytokines, including platelet-derived growth factor, transforming growth factor-beta (TGF beta) and basic fibroblast growth factor (bFGF). Immunohistochemical studies show a close correlation between the mast cell expression of bFGF and the reticulin fibrosis of mastocytosis lesions. The study of cytokine receptor expression also demonstrates that the TGF beta receptor I (RI)-negative cases of mastocytosis are prognostically less favorable than the TGF beta RI-positive cases. This finding may be related to the fact that the TGF beta R complex functions as a tumor suppressor gene in neoplastic cells. PMID- 11919422 TI - Abnormal expression of CD antigens in mastocytosis. AB - Human mast cells are myeloid cells derived from human pluripotential CD34+ stem cells. Normal mast cells exhibit a myeloid immunophenotype characterized by the expression of CD117, CD33 and Fc epsilon RI in the absence of reactivity for CD14, CD15 and lymphoid-lineage-associated antigens. Multiparametric flow cytometric studies have shown that mast cells from mastocytosis display unique immunophenotypic characteristics, including coexpression of CD2 and CD25 antigens together with abnormally high levels of the activation-related antigens CD35, CD63 and CD69 among others. Such aberrant immunophenotypic features are of great relevance in the diagnosis and differential diagnosis of the disease, flow cytometric immunophenotyping of mast cells representing the most sensitive method for the diagnosis of tissue involvement in mastocytosis. From the pathogenetic point of view, the immunophenotypical patterns described suggest the existence of profound changes regarding the adhesion and activation status of mast cells in mastocytosis and may represent a useful tool for a better understanding of some pathophysiological aspects of the disease. PMID- 11919423 TI - Surrogate markers of disease in mastocytosis. AB - Measurement of surrogate mast cell-related products in blood or urine is often performed to assess disease extent in evaluating patients with mastocytosis. Serum tryptase and 24-hour urine histamine metabolites are the most commonly used surrogate markers of mastocytosis. In addition, several novel markers including soluble CD117 and soluble CD25 have been identified in recent studies. The utility and the pitfalls of each of these measurements are discussed. PMID- 11919424 TI - Smouldering mastocytosis: a novel subtype of systemic mastocytosis with slow progression. AB - Systemic mastocytosis (SM) is a clonal disease that shows an either indolent or an aggressive clinical course. Utilizing established criteria, indolent SM can readily be discriminated from the rare aggressive subvariants of SM in most cases. In a small group of patients, however, clinical and laboratory parameters are indicative of slow progression without signs of aggressive disease or an associated hemopoietic neoplasm. These SM patients exhibit a high burden of mast cells, hypercellular marrow and organomegaly. Because of the 'intermediate' course and uncertain prognosis, these cases have been referred to as smouldering SM. In the present article, we discuss clinical and laboratory findings in smouldering SM and review the current literature. In addition, the pathophysiology of this novel subtype of SM is discussed. PMID- 11919425 TI - Spectrum of associated clonal hematologic non-mast cell lineage disorders occurring in patients with systemic mastocytosis. AB - Systemic mastocytosis (SM) is a myeloproliferative disease affecting multipotent and/or mast cell-committed hematopoietic progenitor cells. In a significant subgroup of patients (10-35%), an associated clonal hematologic non-mast cell lineage disorder (AHNMD) occurs. These AHNMDs can be classified according to recently established WHO criteria. Most AHNMDs resemble myeloid malignancies such as acute myeloid leukemia, myeloproliferative disorders or myelodysplastic syndromes. In only a few cases, lymphoproliferative disorders are diagnosed. Patients with SM-AHNMD have a less favorable prognosis concerning survival when compared to indolent SM. No general guidelines for the treatment of patients with SM-AHNMD have been established so far. A reasonable straightforward approach may be to treat the AHNMD in those patients in the same way as if no coexisting SM exists. PMID- 11919426 TI - Cutaneous mastocytosis -- clinical heterogeneity. AB - Cutaneous mastocytosis (CM) encompasses several distinct patterns of disease which can be distinguished by the type of skin lesions, age of onset, familial occurrence, noncutaneous involvement, associated c-kit mutations, and prognosis. A clear distinction of the type of CM in individual patients has important implications regarding patient management and prognosis. PMID- 11919427 TI - Mastocytosis: mediator-related signs and symptoms. AB - Patients with systemic mastocytosis present symptoms related to the tissue response to the release of mediators from mast cells and to the local mast cell burden. Such patients often have a history of chronic and acute mediator-related symptoms. Most patients have indolent disease with a good prognosis and a normal life span. Symptoms can include pruritus, flushing, syncope, gastric distress, nausea and vomiting, diarrhea, bone pain and neuropsychiatric symptoms, most of which are controlled by medication. Because there is no current cure for mastocytosis, successful therapeutic interventions rely on the recognition of mediator-related symptoms and their treatment, and established intervention approaches for the relatively uncommon leukemic concomitants. Efforts to link a particular mast cell-derived mediator to some aspect of the symptom complex depend on the known actions of the mediator and the efficacy of target-based interventions. PMID- 11919428 TI - Mastocytosis: current treatment concepts. AB - An explosion of research on mastocytosis in the last decade has witnessed a greater understanding of the molecular basis of this heterogeneous group of disorders and the conclusion that similar disease phenotypes may indeed be the result of different underlying genotypes. Along with our growing knowledge base of mastocytosis, newer approaches of treating these disorders are becoming available, all under investigational use at this time. This short review highlights the state of the art of current treatment strategies for the different categories of mastocytosis. The future will undoubtedly witness an even greater array of therapeutic options, as we continue to learn more about this enigmatic disease. PMID- 11919429 TI - Treatment of cutaneous mastocytosis. AB - Therapy of cutaneous mastocytosis is directed towards skin and systemic symptoms due to mediator release and targeted on skin lesions. Symptomatic therapy of cutaneous mastocytosis involves agents that inhibit the release of mediators or antagonize H1 and H2 receptors such as antihistamines ketodifen and Aspirin. Disodium cromoglycate has no effect of the cutaneous symptoms of cutaneous mastocytosis. Skin-targeted therapies that lead to a resolution of the lesions of cutaneous mastocytosis are psoralen-photochemotherapy and topical corticosteroid therapy either by occlusion or intralesional injection for a limited number of lesions. There is no treatment that permanently cures cutaneous mastocytosis and patient selection will therefore have to be made on the basis of the clinical manifestations, onset of disease, the probability of spontaneous involution and the severity of cutaneous and systemic symptoms. PMID- 11919430 TI - Laser Vitreolysis. A review. AB - Strands and vitreous adhesion bands can either be dissected noninvasively, transpupillarily by photodisruptive pulses of a Nd:YAG laser, operated in the photodisruptive mode, or invasively using an Er:YAG laser and specialized fibers. The previously used CO(2), Ho:YAG, and ultraviolet lasers have become less popular in the recent past. When using the transpupillary method, specialized contact lenses are required. Noninvasive methods avoid the risks incurred with invasive methods, but they require specialized knowledge, which is not available usually in vitreoretinal services. The invasive laser method provides a number of advantages typical of laser-tissue interaction. Advances in electrosurgical methods have opened the door to a new class of miniaturized electrosurgical equipment with which tissue dissection is made possible by plasma due to dielectrical breakdown which allows the pulse energy to be reduced to a very low level, resulting in a highly localized tissue effect. None of these methods has yet been considered for clinical use, mainly because the presently used mechanical methods are thought to be optimal by the majority of vitreoretinal surgeons. PMID- 11919431 TI - Normal magnetic resonance contrast enhancement of extraocular muscles: a quantitative analysis. AB - The purpose was to evaluate quantitatively the magnetic resonance contrast enhancement of normal extraocular muscles and the use of temporal muscles as a reference of enhancement. Eighty extraocular and 20 temporal muscles were taken into analysis. Before contrast administration, mean intensity of extraocular muscles was found to be higher than that of temporal muscles (p < 0.000). With contrast agent, all extraocular muscles were enhanced more (111% enhancement) than the temporal muscles (45% enhancement, p < 0.000). Lateral recti had the lowest signal intensity, both in pre- and postcontrast images (p < 0.005). Normal extraocular muscles showed prominent enhancement on contrast-enhanced T(1) weighted images. Temporal muscles were also enhanced in all subjects, urging the observers to compare the enhancement of extraocular muscles not with the latter. PMID- 11919432 TI - Typical ultrasound biomicroscopic findings seen in ocular hypotony. AB - Detailed imaging of the ciliary body region by means of ultrasound biomicroscopy can provide information that is crucial for further treatment whereas clinical evaluation and 10-MHz standard B scan are less helpful. BACKGROUND: To evaluate the suitability of ultrasound biomicroscopy (UBM) for elucidating the causes of ocular hypotony. PATIENTS AND METHODS: A retrospective chart review was performed to determine the usefulness of UBM in the evaluation of ocular hypotony. UBM, B scan, and a detailed clinical examination had been performed. Sixty patient charts with prolonged ocular hypotony of different causes were reviewed from January 1994 to December 1998. The information obtained by the UBM was classified into three groups: 'diagnostic' (group 1), 'helpful' (group 2), 'not helpful' (group 3). RESULTS: Of the 60 eyes, 45 UBM examinations (75%) were in group 1, 9 eyes (15%) were assigned to group 2 and 6 eyes (10%) were in group 3. The duration of hypotony had no influence. CONCLUSION: In cases of ocular hypotony, the cause was related to ciliary body pathologies in 80%. Only UBM could distinguish tractional from dehiscence ciliary body detachment which required a different management approach. UBM with its high resolution imaging of the anterior segment - including the ciliary body - is highly suitable for diagnostic clarification. PMID- 11919433 TI - Age-corrected normal values for perimetry. AB - PURPOSE: Age-corrected values are indispensable in assessing discrete changes in light difference sensitivity (LDS) when evaluating perimetric results. In order to detect incipient visual field defects, the present study aimed particularly to determine function of normal values depending on age and eccentricity. METHODS: A test group (n = 83) of ophthalmologically and clinically unremarkable subjects 18 74 years of age was examined with the 'Centerfield' perimeter. A brief introductory program was first used to familiarize each test person with the situation. Then values in the central visual field (30 degrees) were determined with a threshold-splitting program. In addition, the peripheral visual field was examined after a short break. RESULTS: Age-related normal values were calculated both as group averages and as lines of regression by statistical analysis of the resulting data. In contrast to other perimetric studies, decreases in LDS at different test points for persons aged 20 years to persons over 60 were not linear. CONCLUSIONS: Age-related normal values should be reviewed for other perimeters. PMID- 11919434 TI - Re-worsening factor after successful vitrectomy for diabetic retinopathy: optic disc fibrovascular proliferation and macular disease. AB - PURPOSE: To investigate the factors that influence the visual-changing pattern in proliferative diabetic retinopathy even after successful vitrectomy. METHODS: One hundred and forty-seven consecutive eyes were retrospectively reviewed for 6-48 (average 20) months, and were divided into the following 4 groups based on their changing pattern of vision: group A, the visual acuity improved postoperatively and maintained the maximal corrected vision throughout the observation period (n = 49); group B, the visual acuity improved postoperatively but deteriorated thereafter (n = 68); group C, the visual acuity remained the same as before operation (n = 17), and group D, the visual acuity deteriorated immediately after vitrectomy (n = 13). Various issues including systemic conditions, blood tests, preoperative ocular findings, the operative procedures and postoperative complications were reviewed based on the patient records. These issues were analyzed by Spearman's rank correlation, chi(2) test, an analysis of variance (ANOVA) and the Kruskal-Wallis test. Finally, the discriminate factors between groups A and B were examined by a stepwise logistic regression analysis. RESULTS: The following tendencies were observed in all 4 groups: younger patients tended to show a better visual-changing pattern (p = 0.02); patients with younger age at diabetes onset had a better visual-changing pattern after vitrectomy (p = 0.001), and a lower hemoglobin (Hb) A1c level is associated with a better visual changing pattern (p = 0.017). Preoperative rubeosis and macular detachment were frequently found in groups C and D, as well as postoperative rubeosis, vitreous bleeding and retinal detachment. Finally, a stepwise logistic regression analysis showed both fibrovascular proliferation (p = 0.016) from the optic disc and postoperative macular disease (p = 0.0009) to be significant factors for differentiating group A from group B. CONCLUSIONS: In addition to the factors which have already been indicated to affect the visual outcome of a vitrectomy, preoperative findings such as optic disc fibrovascular proliferation and postoperative macular disease were found to affect the visual-changing pattern after a successful vitrectomy. The optimal timing of surgery is very important not only in order to obtain good visual acuity but also to maintain good vision even after a successful vitrectomy. PMID- 11919435 TI - Coagulative, fibrinolytic and metabolic pattern in patients with central retinal vein occlusion. AB - Central retinal vein occlusion (CRVO) is an important cause of visual loss. Many risk factors have been associated with CRVO onset at various ages. Among them diabetes mellitus, hypertension, immunologic disorders, increase in blood viscosity and coagulation, decrease of fibrinolysis have been reported in many subjects. The aim of our study was to detect the metabolic, coagulative and fibrinolytic pattern in 54 patients (26 men, 28 women, mean age 50.4 +/- 12.3) affected by CRVO. We excluded from the study patients with other ocular disorders. A fibrinolytic impairment is the most common feature in our population. It occurs either in dysmetabolic or in nonmetabolic subjects. Such data suggest a prominent role of the fibrinolytic system in the pathogenesis of CRVO. PMID- 11919436 TI - The effects of bupivacaine and lidocaine on the corneal endothelium when applied into the anterior chamber at the concentrations supplied commercially. AB - PURPOSE: To investigate the direct toxic effects of bupivacaine HCl 0.5% and lidocaine HCl 2%, two commonly used injectable local anesthetic agents, on the corneal endothelium when applied intracamerally. METHODS: Two groups were formed, each consisting of 12 pigmented rabbits, and 0.2 ml of the anesthetic agent were injected intracamerally into the right eyes. The central corneal thicknesses and corneal clarities were evaluated preoperatively and at 3, 6, 9, 12 h and 1, 3, 7 days postoperatively. While the central corneal thicknesses were evaluated by ultrasonic pachymetry, the corneal opacification scored between 0 and 3 was assessed by biomicroscopic examination and photographs. RESULTS: Both bupivacaine and lidocaine caused corneal thickening in the 3- to 12-hour measurements. In addition, there was significant corneal opacification in both groups in the 3 hour and 3-day measurements. The corneal thickening and corneal opacification determined during 3- and 6-hour measurements in the eyes which received intracameral bupivacaine were significantly higher than those determined in the lidocaine-injected group. In both groups, the corneal thickness and corneal clarity scores returned to the preoperative values on the 1st and 7th days, respectively. CONCLUSIONS: When applied into the anterior chamber at the concentrations supplied commercially, both bupivacaine and lidocaine cause statistically significant corneal thickening and clinically significant corneal opacification. It should be noted that the injection of these agents into the anterior chamber during the operation at the concentrations supplied commercially may be a potential risk factor for endothelial injury. PMID- 11919437 TI - The effect of artificial tears on corneal surface regularity in patients with Sjogren syndrome. AB - PURPOSE: To evaluate the effect of artificial tears on the topographic parameters associated with corneal surface regularity in patients with Sjogren syndrome. METHODS: A total of 38 eyes of 20 patients with Sjogren syndrome were evaluated prospectively. The mean age of the patients was 50.5 +/- 15.3 years (range, 28 76). Corneal topography with the Topographic Modeling System (TMS-2, Tomey) was performed before and after the instillation of artificial tears. The surface regularity index (SRI), surface asymmetry index (SAI), potential visual acuity (PVA) and mean astigmatism were compared in dry eyes before and after the instillation of artificial tears. RESULTS: Before the instillation of artificial tears SAI, SRI, PVA and mean antigmatism values were found to be 1.37 +/- 1.47, 0.74 +/- 0.42, 20/30.92 +/- 20/8.12 and 1.53 +/- 1.47 D, respectively. In corneal topography performed after the instillation of artificial tears SAI, SRI, PVA and mean astigmatism values were 0.71 +/- 0.82, 0.43 +/- 0.36, 20/25.92 +/- 20/5.55 and 1.08 +/- 1.18, respectively. When compared statistically, the SRI, SAI, and mean astigmatism all decreased significantly and the PVA improved (p = 0.0001). CONCLUSION: Irregular corneal surface in dry eye patients affects optical quality. The statistically significant improvement observed in SRI and SAI values after the instillation of artificial tears also improves the PVA that is especially related to SRI. PMID- 11919438 TI - Effects of timolol and dorzolamide on retrobulbar hemodynamics in patients with newly diagnosed primary open-angle glaucoma. AB - The authors considered a group of patients with newly diagnosed primary open angle glaucoma studying the effects of a 4-week treatment with timolol or dorzolamide on retrobulbar vessels. Ocular hemodynamics were assessed by means of color Doppler imaging of the ophthalmic artery, the temporal short posterior ciliary arteries (SPCAs) and the central retinal artery. For each vessel, systolic and diastolic blood flow velocities were measured, and the resistivity index (RI) was calculated. The only significant result was a reduction of temporal SPCA RI after dorzolamide treatment in comparison with baseline (p = 0.011). In the same group, dorzolamide treatment had a slight and nonsignificant increase in temporal SPCA diastolic velocity. The resistance decrease observed after dorzolamide treatment in the ciliary circulation may be due to the decrease in intraocular pressure or a possible direct vasodilating effect of the drug. PMID- 11919439 TI - Eye health care in the Czech Republic. AB - OBJECTIVE: An analysis of eye health care in the Czech Republic as of 1998 was performed. METHODS: A questionnaire was used to obtain information from all 59 in patient eye departments. RESULTS: The number of ophthalmologists per 1 million inhabitants was 95. The number of cataract operations per 1 million inhabitants was 4,209: phaco-emulsification (36,926 surgeries, 85.2%), extracapsular extraction (6,094 surgeries, 14.1%) and intracapsular extraction (90 surgeries, 0.2%). Intra-ocular lenses were implanted in 99% of cases; 404 corneal transplantations and 1,220 operations for retinal detachment were performed. The number of pars plana vitrectomies for diabetic eye complications was 661. CONCLUSION: Selected regional clinical centres should be equipped and preferred by health insurance companies to provide comprehensive eye health care services and training. PMID- 11919440 TI - In vitro activity of sagamicin against ocular bacterial isolates. AB - The in vitro antibacterial activity of sagamicin, gentamicin, tobramycin and norfloxacin was evaluated against 180 recent clinical isolates obtained from patients with ocular infections. Good activity was demonstrated for the 3 aminoglycosides against methicillin-sensitive Staphylococcus aureus and Staphylococcus epidermidis. All 4 compounds showed a lower activity against methicillin-resistant staphylococci. Sagamicin was highly effective against enterobacteriaceae with a MIC(90) of 2 mg/l and presented good antipseudomonal activity similar to that of gentamicin. Intraocular lenses impregnated with a sagamicin solution showed a good antistaphylococcal activity immediately after preparation. In our strain collection, the ability to produce slime was more frequent among methicillin-resistant S. epidermidis strains than methicillin sensitive S. epidermidis (85 versus 70%). The attachment of 2 S. epidermidis strains to plastic surfaces was partially prevented by sagamicin subinhibitory concentrations. On the contrary, sub-MIC levels of norfloxacin increased the adhesion of S. epidermidis. PMID- 11919441 TI - Oedematous corneal response of the fellow control eye to lotrafilcon a and vifilcon a hydrogel contact lenses in the rabbit. AB - PURPOSE: To compare central corneal swelling after 24 h in eyes wearing Lotrafilcon A (high Dk silicone hydrogel) and Vifilcon A (low Dk hydrogel) lenses and the fellow control eyes of rabbits. METHODS: 24 New Zealand albino rabbits, free of corneal and conjunctival disease, were anaesthetised with ketamine and xylazin. In 12 rabbits, the right eye was fit with the high Dk Lotrafilcon A silicone hydrogel lens. In the other 12 rabbits, the right eye was fit with the low Dk Vifilcon A hydrogel lens, whereas the left eyes were kept as control eyes. Thereafter, the lens-fitted eyes had partial tarsorrhaphy that left a central gap of approximately 4 mm in length. After 24 h the eyelids were opened and the lenses removed. Central corneal thickness was measured using an ultrasonic pachymeter (Mentor O&O-Advent). RESULTS: Central corneal oedema induced by Vifilcon A lens was significantly higher than that with the Lotrafilcon A lens (p < 0.0001). The oedema of the fellow eyes paired with the Vifilcon A lens-wearing eyes was also higher than that of the fellow eyes paired with the Lotrafilcon A lens-wearing eyes (p < 0.01). CONCLUSIONS: The Lotrafilcon A lens induced significantly less corneal oedema than the Vifilcon A lens. Corneal oedema of the fellow eyes appears to be influenced by the swelling of the contralateral lens wearing eyes. The oedema of the fellow control eye was significantly lower when there was less oedema in the contralateral eye wearing a high Dk silicone hydrogel lens. This may be a sympathetic physiological response. The presence of silicone in the high Dk hydrogel lens may suppress corneal oedema in the lens wearing eye, thus affecting the fellow eye. PMID- 11919442 TI - L-carnitine in experimental retinal ischemia-reperfusion injury. AB - The effect of L-carnitine on retinal ischemia-reperfusion injury was evaluated in guinea pigs. 90 min of pressure-induced retinal ischemia followed by 24 h of reperfusion was established in both eyes of 2 groups of animals receiving either L-carnitine (100 mg/kg repeated in 5 doses) or saline intraperitoneally. After enucleation of all the eyes, including those of a control group, malonyldialdehyde (MDA) levels and the thickness of the retinal tissue were measured in 3 groups. The mean MDA value and the tissue thickness of the L carnitine-treated group were statistically insignificant versus the control group (p > 0.05 and p > 0.05, respectively). However, these values were significantly different in the group receiving saline versus the control group and that receiving L-carnitine (p < 0.001, p < 0.001 and p < 0.001, p < 0.001 respectively). L-Carnitine might be an alternative drug for ischemia-reperfusion injury of the retina. PMID- 11919443 TI - Retinal vein occlusion: late complication of a congenital arteriovenous anomaly. PMID- 11919444 TI - Anterior encephaloceles: a study of 92 cases. AB - Anterior encephalocele is a rare condition, and only a few large series have been published in the literature. Surprisingly, the incidence is much higher in Southeast Asian countries, including some parts of India. While the reported incidence in the West is between 1:35,000 and 1:40,000 live births, it is as high as 1:5,000 live births in Thailand. We present a series comprising 92 cases of anterior encephaloceles treated over a 30-year period (1971-2000). Frontoethmoidal encephaloceles are the commonest type, followed by the nasopharyngeal and orbital type. Among the frontoethmoidal encephaloceles, nasoethmoid is the commonest type, and these patients present with swelling over the bridge of the nose with significant hypertelorism and orbital deformities. The nasopharyngeal type remains occult and presents with nasal obstruction or CSF rhinorrhea. Rarely, the patient may present with meningitis. Since 1978, computed tomography (CT) scans have regularly been performed in our patients. CT scans delineate the skull defect and associated brain anomalies. There was associated hydrocephalus present in 12 patients and agenesis of the corpus callosum in 5 patients. In all patients, one-stage repair of the encephalocele and correction of bony anomalies by appropriate osteotomy was undertaken. Since 1988, in cases of frontoethmoidal encephalocele with significant hypertelorism, medial advancement of the medial half of the orbits on either sides was carried out, instead of a classical Tessier's operation. Postoperative morbidity included CSF leak in 20 patients, wound infection in 2 and chest infection in 3. There were 3 deaths in our study. The overall cosmetic outcome was good. PMID- 11919445 TI - Occurrence of split cord malformation in meningomyelocele: complex spina bifida. AB - OBJECTIVE: To describe the clinical features and surgical outcome of a combined anomaly, i.e. split cord malformation (SCM) with meningomyelocele (MMC), and to propose an addition to Pang's classification of SCM to accommodate a combined form of anomaly. METHODS: We retrospectively analyzed 16 cases of such a combination, out of a total of 106 cases of spinal dysraphism treated and studied prospectively for outcome at our center. The clinical profile and outcome of these cases are described. RESULTS: All cases had SCM and MMC. Nine patients were males, and the mean age of presentation was 3.9 years. Twelve patients had Pang's type I SCM and the other 4 were of type II. The MMC sac was lumbar in 11 cases. In all the patients, SCM was present either at the same level as the MMC or one to two segments above it. Nine patients had motor weakness, 6 had hypoesthesia, 4 had urinary incontinence and 3 had trophic ulcers. Nine patients had neuroorthopedic syndrome. All cases, except 3 (who were operated on at birth, at which time 'superficial surgery' was performed without relevant imaging), underwent repair of the MMC and excision of the spur/septum at the same sitting. The 3 cases who had undergone superficial surgery, however, were operated on for SCM following investigation at our center, obviously at a second sitting. After an average follow-up of 7.2 months, 4 patients showed improvement in motor weakness, 5 in hypoesthesia and 3 in urinary symptoms, whereas trophic ulcers had healed in all cases. CONCLUSION: With respect to the occurrence of SCM at or above the level of an MMC, we feel it is apt to screen the entire spinal/neuraxis by MRI in children with MMC. We labeled this combined pathology 'complex spina bifida', and feel it is necessary to make a minor modification to Pang's classification to accommodate the pure/combined anomalies together. PMID- 11919446 TI - Calvarial sclerosing osteomyelitis. AB - We report a 15-year-old boy who suffered from calvarial sclerosing osteomyelitis and presented with painful head swelling. X-rays of the skull revealed areas of irregular radiolucency. MR imaging and CT showed a well-demarcated intradiploic lesion with thickening of the skull extending from the frontal to the parietal calvarium with a low signal on T1-weighted images, strong but heterogeneous enhancement after gadolinium application and a mixed signal on T2-weighted images. Computer-navigated neurosurgery was planned, and the craniotomy defect was reconstructed by a preformed titanium implant. Sclerosing osteomyelitis of the calvarium has to be included in the differential diagnosis of osteolytic and sclerosing lesions of the skull coinciding with persistent swelling of the head. PMID- 11919447 TI - Impact of selective posterior rhizotomy on fine motor skills. Long-term results using a validated evaluative measure. AB - Suprasegmental effects following selective posterior rhizotomy have been frequently reported. However, few studies have used validated functional outcome measures to report the surgical results beyond 3 years. The authors analyzed data obtained from the McGill Rhizotomy Database to determine the long-term impact of lumbosacral dorsal rhizotomy on fine motor skills. The study population comprised children with debilitating spasticity who underwent SPR and were evaluated by a multidisciplinary team preoperatively, at 6 months and 1 year postoperatively. Quantitative standardized assessments of upper extremity function were obtained using the fine motor skills section of the Peabody Developmental Motor Scales (PDMS) test. Of 70 patients who met the entry criteria for the study, 45 and 25 completed the 3- and 5-year assessments, respectively. Statistical analysis demonstrated significant improvements in grasping, hand use, eye-hand coordination, and manual dexterity at 1 year after SPR. More importantly, all improvements were maintained at 3 and 5 years following SPR. This study supports that significant improvements in upper extremity fine motor function using the PDMS evaluative measure are present after SPR and that these suprasegmental benefits are durable. PMID- 11919448 TI - Comparison of motor outcomes after selective dorsal rhizotomy with and without preoperative intensified physiotherapy in children with spastic diplegic cerebral palsy. AB - A previous randomized clinical trial compared selective dorsal rhizotomy (SDR) plus postoperative intensified physiotherapy (group 1) with intensified physiotherapy alone (group 2) for children with spastic diplegic cerebral palsy. At the end of this trial, all patients in group 2 had an SDR, followed by further intensified physiotherapy. This study was performed to determine if the additional intensified physiotherapy before SDR, as occurred in group 2, improved long-term motor outcome. Outcomes were compared in the two groups, i.e. group 1 without intensified physiotherapy before SDR and group 2 with intensified physiotherapy before SDR. The primary outcome measure was the Gross Motor Function Measure (GMFM). Lower-limb spasticity, range and strength were secondary outcome measures. Baseline assessments had been done for the prior clinical trial. For this study, patients were reassessed by physiotherapists blinded to the treatment group. Thirteen children in each group were studied at a mean follow-up of 53 months. The mean improvement in GMFM was 10.0 in group 1 and 10.4 in group 2 (p = 0.9). Improvements in spasticity and range were similar in the two groups. There was no significant change in muscle strength in either group. It was concluded that in this relatively small series of patients, additional intensified physiotherapy before SDR did not improve motor outcomes. PMID- 11919449 TI - Choroid plexectomy reduces neurosurgical intervention in patients with hydranencephaly. AB - INTRODUCTION: The removal of the choroid plexus from the lateral ventricles was attempted by Dandy in the early 20th century but later discarded as complications arose and other methods of cerebrospinal fluid (CSF) diversion were introduced. We compare our experience with a variation of this operation to CSF diversion in patients with hydranencephaly or near hydranencephaly. METHODS: The hospital and office charts of patients with a diagnosis of hydranencephaly were reviewed from the two institutions spanning the career of the senior author. Thirteen patients were identified, of whom 9 underwent CSF diversionary procedures (group A) and 4 underwent choroid plexectomy (group B). RESULTS: The mean number of reoperations (2 in group A, 0 in group B), neurosurgical readmissions (1.5 in group A, 0 in group B) and days of hospitalization related to neurosurgical readmissions (43.5 in group A, 0 in group B) were all less in patients who underwent choroid plexectomy. The total incidence of complications related to surgery was also lower in this group (7 in group A, 0 in group B). CONCLUSION: In our experience, choroid plexectomy in patients with hydranencephaly reduces the incidence of reoperation and readmission, the number of days of hospitalization related to the surgical procedure and the total number of complications in comparison to patients undergoing CSF diversion. Further neurosurgical intervention is minimized as is the financial burden from multiple emergency department visits and radiological procedures for shunt evaluation. Choroid plexectomy is a viable alternative to CSF diversion in patients with hydranencephaly and a rapidly enlarging head. It avoids the chronic issues and complications surrounding CSF diversion in this difficult group of patients. PMID- 11919450 TI - Schwannoma of the tentorium cerebelli in a child. Case report. AB - Intracranial schwannomas not arising from a cranial nerve are very rare. Schwannomas of the dura are even rarer; in the literature, we found only two cases: a schwannoma of the falx and one of the torcula. We report a third case of a 9-year-old girl with a schwannoma of the tentorium cerebelli. The different theories concerning the origin of this tumor in this particular location are discussed. PMID- 11919451 TI - Moyamoya disease associated with hemophilia A. Case report. AB - A 10-year-old boy who had been diagnosed as having hemophilia A presented with episodes of transient ischemic attack. Cerebral angiography showed occlusions of the bilateral anterior cerebral arteries and the right middle cerebral artery with the development of an abnormal vascular network. The patient was diagnosed as having moyamoya disease associated with hemophilia A and subsequently underwent multiple burr hole surgery for revascularization under sufficient factor VIII supplementation. He remained asymptomatic after surgery, and follow up cerebral angiography 5 months after the operation demonstrated significant neovascularization through the burr holes. Multiple burr hole surgery is proposed as a method of choice for the treatment of moyamoya disease associated with a bleeding tendency. PMID- 11919452 TI - Visual pathway glioma. PMID- 11919453 TI - Lumbar dermoid cyst causing pyomyelia in a child. PMID- 11919454 TI - Interscapular placement of a vagal nerve stimulator pulse generator for prevention of wound tampering. Technical note. AB - In some cognitively delayed children who require a vagal nerve stimulator for treatment of their seizures, there is a risk of wound breakdown and infection from obsessive tampering with the wound. We describe the interscapular placement of the vagal nerve stimulator pulse generator as a method to reduce this risk. PMID- 11919455 TI - Repeat cerebrospinal fluid shunt infection: recurrence, relapse, repeat or reinfection? PMID- 11919456 TI - Chronic pelvic pain in men. AB - Chronic pelvic pain is a condition which receives less attention in men than in women. It is often difficult to diagnose and more difficult to treat. The new classification of prostatis and its variants has introduced the term 'chronic pelvic pain syndrome' which underlines the difficulties in dealing with this disorder which may represent a variety of chronically painful conditions with a large functional component. PMID- 11919457 TI - Plasma antidiuretic hormone levels in children with spina bifida. AB - PURPOSE: Urological management of spina bifida patients is controversial. The goals of therapy of neurogenic bladder are continence, prevention of infections and preservation of urinary tract. Desmopressin has been recently used in a spina bifida population that is dry during the day (daytime continence was achieved with clean intermittent catheterization and anticholinergics) but wet at night. The aim of this study was to assess plasma antidiuretic hormone (ADH) levels in these children. MATERIALS AND METHODS: The study included 24 patients, 11 males and 13 females (mean age 6.4 years) referred to the Spina Bifida Centre of the Catholic University of Rome, and 57 normal age-matched controls. Morning (07.30 08.00 h) plasma ADH levels were measured using a specific radioimmunoassay. RESULTS: Plasma ADH levels (normal range 5-11 microg/l) did not differ between spina bifida population and healthy controls. Serum ADH had a mean of 6.8 microg/l in affected children and a mean of 7.4 microg/l in the controls. CONCLUSION: We conclude that the use of desmopressin in children with spina bifida should be reserved only in patients with decreased secretion of ADH, or may be useful in patients with persistent nocturnal incontinence to reduce night wetting. Therefore, research with a larger population is needed. PMID- 11919458 TI - Complications of Stamey needle suspension for female stress urinary incontinence. AB - INTRODUCTION: The Stamey bladder neck suspension for stress urinary incontinence in females is thought to be an excellent procedure. However, recent studies revealed that complications of this procedure have not been negligible. In this retrospective study, complications of the Stamey needle bladder neck suspension were examined. MATERIALS AND METHODS: The Stamey procedure was performed either alone or in combination with anterior colporrhaphy for 86 female patients with stress urinary incontinence between 1989 and 1999. The mean follow-up period was 37.6 months and the mean age was 59.1 years. We studied the complications postoperatively pointed out by patients' complaint and image examination. RESULTS: The overall incidence of complications was 37.2%. Voiding difficulties and lower abdominal pain were present in 15 cases, respectively. We experienced 2 unusual cases in whom suspensory stitches had to be removed due to abdominal pain. CONCLUSIONS: Although the Stamey bladder neck suspension is thought to be an excellent procedure for stress urinary incontinence, surgeons should beware of possible complications and be alert of the symptoms, such as abdominal pain and bladder irritation. PMID- 11919459 TI - Relation between volume of residual urine and urodynamic findings in women with cystourethrocele. AB - OBJECTIVE: To evaluate the relation between post-voiding residual urine (PVR) and urodynamic findings, voiding patterns, and the degree of prolapse in women with cystourethrocele. PATIENTS AND METHODS: According to the average PVR value measured by means of catheterization after three separate and spontaneous micturitions, 73 women with minimal urethral hypermobility (n = 8) or colpocystocele (n = 65) were subdivided into 3 groups: group 1 = patients with a PVR of < or =50 ml; group 2 = patients with a PVR of >50 ml and < or =150 ml, and group 3 = patients with a PVR of >150 ml and < or =500 ml. The patients underwent a thorough physical examination, and prolapse was evaluated and graded. The women then underwent a complete urodynamic investigation. The 3 groups of patients were statistically correlated according to the urodynamic findings, voiding patterns and degree of prolapse. RESULTS: A large PVR in women with cystourethrocele was often associated with poor maximum flow rate, intermittent flow and with the highest degree of prolapse. CONCLUSION: The present study suggests that uroflowmetry is a very simple, noninvasive, but also essential method of investigation in evaluating female patients with cystourethrocele, particularly when a large PVR is suspected. PMID- 11919460 TI - Diagnostic value of 18F-FDG positron emission tomography for detection and treatment control of malignant germ cell tumors. AB - INTRODUCTION: The role of positron emission tomography (PET) with 2-[18F]fluoro-2 deoxy-D-glucose ([18F]FDG) is currently under evaluation in urologic oncology. The aim of the present study was to investigate the use of [18F]FDG positron emission tomography ([18F]FDG-PET) in the detection and treatment control of malignant germ cell tumors compared to computed tomography (CT). MATERIALS AND METHODS: Thirty-two PET studies and CT scans were carried out in 23 patients with histologically proven germ cell tumors (10 seminomas, 12 non-seminomatous germ cell tumors (NSGCT), 1 unclassified serologic recurrent disease) Lugano stage I III. The scans were done either after initial diagnosis (n = 21) and/or within 3 45 days after chemotherapy was completed (n = 11). PET and CT were validated either by histology (n = 7) or clinical follow-up of 6-11 months after the last PET study has been performed (n = 16). Sensitivity, specificity, accuracy, positive and negative predictive values were determined for PET and CT. Differences between PET and CT for parameters of diagnostic value were evaluated by chi(2) test. RESULTS: Although not statistically significant, the sensitivity, accuracy and negative predictive value were higher for PET than for CT with respect to the detection of metastatic infradiaphragmatic and supradiaphragmatic lesions after initial diagnosis. The specificity and positive predictive value of PET and CT were comparable. After chemotherapy, PET was found to be significantly superior in specificity and accuracy compared to CT with respect to infradiaphragmatic lesions (p < 0.05). False-positive PET findings in supradiaphragmatic lesions after chemotherapy occurred in the case of inflammatory processes and resulted in a loss of specificity and accuracy compared to CT (p < 0.05). CONCLUSIONS: These preliminary results demonstrate [18F]FDG-PET to be a useful diagnostic tool for the initial staging and treatment control in patients with germ cell tumors. Possible advantages compared to CT, however, are as yet not clearly defined. The possibility of false-positive PET findings due to reactive supradiaphragmatic inflammatory processes early after chemotherapy have to be considered. PMID- 11919461 TI - Effect of the rate of delivery of lignocaine gel on patient discomfort perception prior to performing flexible cystoscopy. AB - Flexible cystoscopy is commonly performed under local anaesthesia. Instillation of lignocaine gel is commonly associated with urethral discomfort, which in some cases results in fierce opposition to further flexible cystoscopy under local anaesthesia. Although studies have demonstrated that the temperature of lignocaine can influence the level of discomfort experienced, to date no study has investigated the influence of the rate of lignocaine delivery on perceived discomfort. We therefore performed a prospective, randomised study to investigate this in patients undergoing flexible cystoscopy. One hundred consenting men were randomised to receive 11 ml of 2% lignocaine hydrochloride gel over either 2 or 10 s. The groups were well matched for age. After instillation of the gel, the patients were immediately asked to score their discomfort using a visual analogue scale. The discomfort experienced by patients that received the gel over 10 s was significantly (p < 0.05; Student's t test) less than those that received it over 2 s. This was irrelevant of the age of the patient and the number of previous cystoscopies performed. We have demonstrated that slow administration results in decreased discomfort. This may, in turn, reduce the need to resort to general anaesthesia, which is associated with increased morbidity and cost. PMID- 11919462 TI - Renography as a measure of renal and neobladder function in patients with ileal bladder substitution. AB - INTRODUCTION: Using renography, we evaluated the function of both the kidneys and neobladder of 20 patients who underwent ileal bladder substitution. METHODS: The patients were measured for renal and neobladder function with renography using 99mTc MAG3. Renal function was assayed in every renal unit using O'Reilly patterns. Voiding function was estimated using radioisotope counts for the reservoir. RESULTS: Thirty-two of 37 renal units had normal renograms. Five renal units showed non-obstructive renograms. The percentage of total residual urine (RU) in the reservoir was 30% and more in 6 patients, 10-30% in 8, and <10% in 6. In 7 of 12 patients, the data on estimated RU correlated with the data on measured RU. However, in 5 patients, the data on estimated RU increased more in quantity than the data on RU measured at another time. CONCLUSIONS: In patients with bladder substitution the functions of both the kidneys and neobladder could be evaluated in a single renography. PMID- 11919463 TI - Epidemiological studies on the prevalence of upper urinary calculi in Taiwan. AB - INTRODUCTION: A nationwide survey was conducted to investigate the prevalence of upper urinary calculi in Taiwan. MATERIALS AND METHODS: A postal questionnaire was mailed to 27,758 people, 0.2% of the adults in Taiwan. RESULTS: Of the 4,588 valid respondents, 440 had at least one episode of upper urinary calculus disease. The overall prevalence was 9.6% (14.5% in males and 4.3% in females). Men were more prone to nephrolithiasis than women (age-adjusted prevalence of 12.2% in men and 3.1% in women, p = 0). The 'stone belt' was localized in the Midwest region of Taiwan. A gender- and age-adjusted multivariate analysis revealed that alcohol consumption and family history of kidney stone were significant risk factors for stone occurrence. Compared with general population, the odds ratios for stone disease in inhabitants whose father, mother and both parents with stone history were 3.44 [95% confidence interval (CI), 2.33-5.07], 4.79 (95% CI, 2.85-8.07) and 10.40 (95% CI, 3.75-28.84), respectively. The subtropical temperature and gradually higher socioeconomic standards of living may contribute to the high prevalence. Inhabitants in the Midwest region have higher risk to develop stones. CONCLUSIONS: Further studies are needed to investigate the exact cause of these regional variations of stone prevalence. Nevertheless, the present study provides an additional piece of information on worldwide epidemiology of urolithiasis. PMID- 11919464 TI - Are clinical characteristics of familial benign prostatic hyperplasia different than in sporadic cases? AB - OBJECTIVE: Familial benign prostatic hyperplasia (BPH) is a recently popularized entity with yet uncertain clinical and pathological features. In the present study we investigated whether there was any difference between clinical characteristics of familial and sporadic BPH in a series of 148 surgically treated BPH patients. MATERIALS AND METHODS: A retrospective analysis was performed in 148 patients subjected to transvesical or transurethral prostatectomy to determine the clinical features of familial BPH. Patients were categorised as having familial BPH when 3 or more (including the patient) first degree family members gave a history of BPH. Accordingly 23 cases who fit this criterion were accepted as having familial BPH and the rest of the cases were taken as the control group. The two groups were compared with respect to age, International Prostate Symptom Score (IPSS), quality of life score, prostate specific antigen (PSA), maximum urinary flow rate and the weight of the surgical prostate specimen. RESULTS: The mean age, IPSS, quality of life score, total PSA, maximum urinary flow rate and the weight of the surgical prostate specimen were found as 65.13 +/- 5.51 years, 23.13 +/- 4.82, 4.78 +/- 0.95, 6.0 +/- 4.1 ng/ml, 6.9 +/- 2.7 ml/s and 62.96 +/- 38.76 g, respectively, in the familial BPH group whereas the same parameters were measured as 68.13 +/- 7.68 years, 24.74 +/- 3.73, 4.52 +/- 0.85, 5.93 +/- 4.75 ng/ml, 4.6 +/- 1.71 ml/s and 70.87 +/- 53.21 g, respectively. No significant difference was present between familial and sporadic BPH cases in any of the studied parameters. CONCLUSION: The clinical features of familial BPH did not differ significantly from those of sporadic BPH. PMID- 11919465 TI - Comparative study of peptides isolated from class I antigen groove of urological specimens. a further step toward the future of peptide therapy in bladder cancer patients. AB - BACKGROUND: The major histocompatibility (MHC) class I antigens act as associative molecules for interaction amongst immuno-competent cells. The grooves of class I antigens are normally loaded with self peptides of between 8 and 11 amino acids. However, when the cells transform to malignant state they may carry peptide(s) of non-self origin within these grooves. Using immuno-bead purification followed by high-performance liquid chromatography (HPLC), this study attempted to isolate peptides from class I antigens of various biological specimens RESULTS: The combination of immuno-bead purification (BP) and HPLC was reliable for peptide isolation. Class I antigens and associated peptides could be isolated from normal peripheral blood leukocyte (PBL). Under the same conditions, the PBL yielded almost twice as much peptide as that of tumour cell lines. The HPLC profile of peptides (range of 8-10 amino acid residues) isolated from a bladder and a cervical tumour cell line showed unique features. In the case of the bladder line there were at least 22 peptides. In addition, the class I associated peptides could also be isolated from kidney tumour fragments of three individuals. In each the isolated peptides showed a unique HPLC peak profile with some similarities as well as differences. CONCLUSIONS: These data indicated a variation in the nature of peptides isolated from different specimens. The approach showed the feasibility of preparing peptide(s) from a relatively small number of cells. The data also showed that peptide isolation could also be carried out from tumour tissue biopsies paving the way for the future of peptide vaccination in cancer patients following the identification of putative tumour specific replicate(s). PMID- 11919466 TI - Detection of LOH of the RB1 gene in bladder cancers by PCR-RFLP. AB - OBJECTIVES: Retinoblastoma (RB1) gene involves in retinoblastoma, osteosarcoma, bladder, prostate, lung, breast carcinomas, and soft tissue sarcomas. Loss of heterozygosity (LOH) is the most common mutation of the gene. METHODS: Xba I polymorphism in intron 17 of the gene was used to detect LOH in 20 bladder cancer patients. A cystitis and an osteosarcoma were used as control. LOH was investigated in three different kinds of samples (blood, paraffin-embedded tissue and fresh tissue) belonging to the same patients, and 20 blood samples, 20 paraffin-embedded tissue samples and 16 fresh tissue samples were obtained from 20 cancer patients. RESULTS: None of the 20 blood samples showed LOH. Eleven out of 20 paraffin-embedded bladder tissues were amplified, 3 of them homozygous and all 8 informative paraffin-embedded tissues showed LOH. Five out of 16 fresh tumor tissues obtained were amplified, in 1 the fresh tissue was normal, 1 fresh tissue showed LOH and 3 were not digested by Xba I. CONCLUSION: The results of the study have suggested that detection of LOH of the RB1 gene by PCR-RFLP can be a good adjunctive test for evaluation of the bladder cancer. PMID- 11919467 TI - Study of efferent tube suspension as a continent diversion mechanism. an experimental study in dogs. AB - PURPOSE: To construct a reliable continent tube, which is easy to catheterize and surgically simple. MATERIALS AND METHODS: 8 adult female mongrel dogs underwent a procedure in which an ileal segment was tapered as an efferent tube. The tapered ileum was fixed to the back surface of the rectus muscle with a polyester taper 1.5 cm in width and a suspension tension of 400 g. The internal orifice of the tapered ileum was anastomosed to the bladder and the external orifice of the tapered ileum was brought out to the abdominal skin. Urodynamic and radiological studies were carried out postoperatively in all dogs. RESULTS: All stomas in dogs could be easily catheterized with a 14-french catheter. Urodynamic study of the efferent tubes showed that the maximum closure pressure ranged from 117 to 157 (mean 136.75) cm water. Retrograde radiogram of the efferent tubes showed perfect canalization without stenosis. CONCLUSION: This study suggests that the continent mechanism of the tapered ileum can be greatly enhanced by a suspension technique. PMID- 11919468 TI - Strangulation of the penis by a self-circumcision device. AB - Penile strangulation caused by wearing metallic or non-metallic objects is unusual but potentially serious. We report the case of a 33-year-old man who presented with penile strangulation after applying a self-circumcision device. The device was removed, and after conservative treatment conventional circumcision was performed. It is stressed that circumcision should be performed by urologists or medically trained personnel. PMID- 11919469 TI - Recurrence of renal cell carcinoma with extensive vena caval thrombus three years after radical nephrectomy. AB - A 70-year-old man, who had undergone a radical nephrectomy for localized renal cell carcinoma (RCC) three years ago, was recently evaluated for shortness of breath. During his work-up, a transesophageal echocardiogram and magnetic resonance imaging indicated an extensive vena caval thrombus originating from the renal vein stump. Successful vena caval thrombectomy with cardiopulmonary bypass confirmed that the thrombus was comprised of RCC. This is a rare type of RCC recurrence with an unusual timing (3 years after a nephrectomy), alerting the importance of strict follow-up on all patients even after complete surgical excision of all suspected tumors. PMID- 11919470 TI - Renal cell carcinoma presenting as acute pancreatitis. AB - We present a 50-year-old male with renal cell carcinoma and hypercalcemia presenting as acute pancreatitis. The sudden onset of epigastralgia, acute pancreatitis and hypercalcemia with raised parathyroid-like peptide levels in this patient could not be explained except for renal cell carcinoma. This is the first report of such a complication in renal cell carcinoma. The rare presentation and management with a review of the literature is described. PMID- 11919471 TI - Rare secondary carcinoma from bladder to testis. AB - Metastatic carcinoma to the testis is very unusual in daily urologic practice. We report a case of metastatic cancer to the testis detected as incidental findings in a squamous bladder tumor. PMID- 11919472 TI - Opsoclonus-myoclonus syndrome as a paraneoplastic manifestation of renal cell carcinoma. a case report and review of the literature. AB - Opsoclonus is a rare but distinctive disorder of ocular motility, characterized by irregular, continual and conjugated chaotic saccades of the eyes. It is increased with eye closure and fixation, and it persists during sleep. Opsoclonus appears more commonly in children and in half of such cases in association with neuroblastoma. In adults the most frequent causes are idiopathic (50%) and paraneoplastic (20%). Among the second group, different types of tumors involving a wide variety of organs have been reported. Opsoclonus when accompanied by other symptoms of central nervous system involvement (head, appendicular myoclonus and truncal ataxia) constitutes a clinical picture, known as opsoclonus-myoclonus syndrome. We report the case of a young patient affected by renal cell carcinoma associated with opsoclonus-myoclonus syndrome which did not respond to medical therapy but dramatically improved after cancer ablation. PMID- 11919473 TI - Hepatic arterioportal shunts: dynamic CT and MR features. AB - With the increased temporal resolution available in dynamic computed tomography (CT) and magnetic resonance imaging (MRI), hepatic arterioportal shunts are now more frequently encountered than in the past. The condition occurs in various hepatic diseases in which portal or hepatic venous flow is compromised. The underlying mechanism and the degree of shunt affect its appearance at dynamic imaging. The dynamic CT and MRI findings have been summarized as early enhancement of peripheral portal veins, and wedge-shaped transient parenchymal enhancement during the hepatic arterial phase. Recognition of arterioportal shunt can suggest the presence of a previously unsuspected disorder and avoids false positive diagnosis or overestimation of a hepatic disease. Familiarity with the pathophysiology of arterioportal shunt also allows investigation of the hepatic hemodynamic changes occurring in various hepatic diseases. PMID- 11919474 TI - The usefulness of MR imaging of the temporal bone in the evaluation of patients with facial and audiovestibular dysfunction. AB - OBJECTIVE: To evaluate the clinical utility of MR imaging of the temporal bone in patients with facial and audiovestibular dysfunction with particular emphasis on the importance of contrast enhancement. MATERIALS AND METHODS: We retrospectively reviewed the MR images of 179 patients [72 men, 107 women; average age, 44 (range, 1-77) years] who presented with peripheral facial palsy (n=15), audiometrically proven sensorineural hearing loss (n=104), vertigo (n=109), or tinnitus (n=92). Positive MR imaging findings possibly responsible for the patients' clinical manifestations were categorized according to the anatomic sites and presumed etiologies of the lesions. We also assessed the utility of contrast-enhanced MR imaging by analyzing its contribution to the demonstration of lesions which would otherwise not have been apparent. All MR images were interpreted by two neuroradiologists, who reached their conclusions by consensus. RESULTS: MR images demonstrated positive findings, thought to account for the presenting symptoms, in 78 (44%) of 179 patients, including 15 (100%) of 15 with peripheral facial palsy, 43 (41%) of 104 with sensorineural hearing loss, 40 (37%) of 109 with vertigo, and 39 (42%) of 92 with tinnitus. Thirty (38%) of those 78 patients had lesions that could be confidently recognized only at contrast-enhanced MR imaging. CONCLUSION: Even though its use led to positive findings in less than half of these patients, MR imaging of the temporal bone is a useful diagnostic procedure in the evaluation of those with facial and audiovestibular dysfunction. Because it was only at contrast-enhanced MR imaging that a significant number of patients showed positive imaging findings which explained their clinical manifestations, the use of contrast material is highly recommended. PMID- 11919475 TI - Dependent lung opacity at thin-section CT: evaluation by spirometrically-gated CT of the influence of lung volume. AB - OBJECTIVE: To evaluate the influence of lung volume on dependent lung opacity seen at thin-section CT. MATERIALS AND METHODS: In thirteen healthy volunteers, thin-section CT scans were performed at three levels (upper, mid, and lower portion of the lung) and at different lung volumes (10, 30, 50, and 100% vital capacity), using spirometric gated CT. Using a three-point scale, two radiologists determined whether dependent opacity was present, and estimated its degree. Regional lung attenuation at a level 2 cm above the diaphragm was determined using semiautomatic segmentation, and the diameter of a branch of the right lower posterior basal segmental artery was measured at each different vital capacity. RESULTS: At all three anatomic levels, dependent opacity occurred significantly more often at lower vital capacities (10, 30%) than at 100% vital capacity (p = 0.001). Visually estimated dependent opacity was significantly related to regional lung attenuation (p < 0.0001), which in dependent areas progressively increased as vital capacity decreased (p < 0.0001). The presence of dependent opacity and regional lung attenuation of a dependent area correlated significantly with increased diameter of a segmental arterial branch (r = 0.493 and p = 0.0002; r = 0.486 and p = 0.0003, respectively). CONCLUSION: Visual estimation and CT measurements of dependent opacity obtained by semiautomatic segmentation are significantly influenced by lung volume and are related to vascular diameter. PMID- 11919476 TI - Tubular foreign body or stent: safe retrieval or repositioning using the coaxial snare technique. AB - OBJECTIVE: To evaluate the utility and advantages of the coaxial snare technique in the retrieval of tubular foreign bodies. MATERIALS AND METHODS: Using the coaxial snare technique, we attempted to retrieve tubular foreign bodies present in seven patients. The bodies were either stents which were malpositioned or had migrated from their correct position in the vascular system (n=2), a fragmented venous introducer sheath (n=1), fragmented drainage catheters in the biliary tree (n=2), or fractured external drainage catheters in the urinary tract (n=2). After passing a guidewire and/or a dilator through the lumina of these foreign bodies, we introduced a loop snare over the guidewire or dilator, thus capturing and retrieving them. RESULTS: In all cases, it was possible to retrieve or reposition the various items, using a minimum-sized introducer sheath or a tract. No folding was involved. In no case were surgical procedures required, and no complications were encountered. CONCLUSION: The coaxial snare technique, an application of the loop snare technique, is a useful and safe method for the retrieval of tubular foreign bodies, and one which involves minimal injury to the patient. PMID- 11919477 TI - Sonographic evaluation of breast nodules: comparison of conventional, real-time compound, and pulse-inversion harmonic images. AB - OBJECTIVE: To compare the use of conventional, real-time compound, and pulse inversion harmonic imaging in the evaluation of breast nodules. MATERIALS AND METHODS: Fifty-two breast nodules were included in this study, conducted between May and December 2000, in which conventional, real-time compound, and pulse inversion harmonic images were obtained in the same plane. Three radiologists, each blinded to the interpretations of the other two, evaluated the findings, characterizing the lesions and ranking the three techniques from grade 1, the worst, to grade 3, the best. Lesion conspicuity was assessed, and lesions were also characterized in terms of their margin, clarity of internal echotexture, and clarity of posterior echo pattern. The three techniques were compared using Friedman's test, and interobserver agreement in image interpretation was assessed by means of the intraclass correlation coefficient. RESULTS: With regard to lesion conspicuity, margin, and internal echotexture of the nodules, real-time compound imaging was the best technique (p < 0.05); in terms of posterior echo pattern, the best was pulse-inversion harmonic imaging (p < 0.05). Real-time compound and pulse inversion harmonic imaging were better than conventional sonography in all evaluative aspects. Interobserver agreement was greater than moderate. CONCLUSION: Real-time compound and pulse-inversion harmonic imaging procedures are superior to conventional sonography in terms of both lesion conspicuity and the further characterization of breast nodules. Real-time compound imaging is the best technique for evaluation of the margin and internal echotexture of nodules, while pulse-inversion harmonic imaging is very effective for the evaluation of the posterior echo patterns. PMID- 11919478 TI - The usefulness of pulsatile flow detection in measuring resistive index in renal Doppler US. AB - OBJECTIVE: To assess the usefulness of pulsatile flow detection (PFD), a newly developed function of color Doppler US, in measuring resistive index (RI) in renal Doppler US and to compare it with conventional color Doppler (CCD). MATERIALS AND METHODS: Fifty-six kidneys in 31 patients were randomly selected and divided into two groups. In group A, RI was measured first with the aid of CCD, and then with PFD. In group B, data were obtained in the reverse order. The time required for each RI measurement was recorded in seconds. The quality of the Doppler spectral waveform was subjectively graded as 0, 1, or 2 and examination time and waveform quality were compared between PFD and CCD. RESULTS: The time required to measure RI with PFD (PFD time) was less than with CCD (CCD time) (mean 42.7 secs vs. mean 70.3 secs; p = 0.031). There was no significant difference in PFD time between group A and B, but CCD time was shorter in group B (70.3 secs vs. 24.6 secs; p = 0.0004). Spectral waveform quality was not significantly different between PFD and CCD. CONCLUSION: The time required to measure RI in kidneys can be shortened with the aid of the PFD function in color Doppler US without affecting the quality of the examination. PMID- 11919479 TI - The quality of reconstructed 3D images in multidetector-row helical CT: experimental study involving scan parameters. AB - OBJECTIVE: To determine which multidetector-row helical CT scanning technique provides the best-quality reconstructed 3D images, and to assess differences in image quality according to the levels of the scanning parameters used. MATERIALS AND METHODS: Four objects with different surfaces and contours were scanned using multidetector-row helical CT at three detector-row collimations (1.25, 2.50, 5.00 mm), two pitches (3.0, 6.0), and three different degrees of overlap between the reconstructed slices (0%, 25%, 50%). Reconstructed 3D images of the resulting 72 sets of data were produced using volumetric rendering. The 72 images were graded on a scale from 1 (worst) to 5 (best) for each of four rating criteria, giving a mean score for each criterion and an overall mean score. Statistical analysis was used to assess differences in image quality according to scanning parameter levels. RESULTS: The mean score for each rating criterion, and the overall mean score, varied significantly according to the scanning parameter levels used. With regard to detector-row collimation and pitch, all levels of scanning parameters gave rise to significant differences, while in the degree of overlap of reconstructed slices, there were significant differences between overlap of 0% and of 50% in all levels of scanning parameters, and between overlap of 25% and of 50% in overall accuracy and overall mean score. Among the 18 scanning sequences, the highest score (4.94) was achieved with 1.25 mm detector-row collimation, 3.0 pitch, and 50% overlap between reconstructed slices. CONCLUSION: Comparison of the quality of reconstructed 3D images obtained using multidetector row helical CT and various scanning techniques indicated that the 1.25 mm, 3.0, 50% scanning sequence was best. Quality improved as detector-row collimation decreased; as pitch was reduced from 6.0 to 3.0; and as overlap between reconstructed slices increased. PMID- 11919480 TI - Radiological spectrum of intraductal papillary tumors of the bile ducts. AB - Papillary tumor of the bile duct is characterized by the presence of an intraductal tumor with a papillary surface comprising innumerable frondlike infoldings of proliferated columnar epithelial cells surrounding slender fibrovascular stalks. There may be multiple tumors along the bile ducts (papillomatosis or papillary carcinomatosis), which are dilated due to obstruction by a tumor per se, by sloughed tumor debris, or by excessive mucin. Radiologically, the biliary tree is diffusely dilated, either in a lobar or segmental fashion, or aneurysmally, depending on the location of the tumor, the debris, and the amount of mucin production. A tumor can be depicted by imaging as an intraductal mass with a thickened and irregular bile duct wall. Sloughed tumor debris and mucin plugs should be differentiated from bile duct stones. Cystically or aneurysmally, dilated bile ducts in mucin-hypersecreting variants (intraductal papillary mucinous tumors) should be differentiated from cystadenoma, cystadenocarcinoma and liver abscess. PMID- 11919481 TI - Prenatal ultrasound findings of fetal neoplasms. AB - A variety of neoplasms can develop in each fetal organ. Most fetal neoplasms can be detected by careful prenatal ultrasonographic examination. Some neoplasms show specific ultrasonographic findings suggesting the differential diagnosis, but others do not. Knowledge of the presence of a neoplasm in the fetus may alter the prenatal management of a pregnancy and the mode of delivery, and facilitates immediate postnatal treatment. During the last five years, we experienced 32 cases of fetal neoplasms in a variety of organs. We describe their typical ultrasonographic findings with correlating postnatal CT, MRI, and pathologic findings. PMID- 11919482 TI - Spontaneous partial regression of cerebral arteriovenous malformation. AB - Arteriovenous malformation (AVM) of the brain is one of the important pathologic conditions which cause intracerebral or subarachnoid hemorrhage, epilepsy, or chronic cerebral ischemia. The spontaneous regression of cerebral AVM is reported to be very rare and more likely to occur when the AVM is small, is accompanied by hemorrhage, and has fewer arterial feeders. We report a case of right occipital AVM which at follow-up angiography performed four years later showed near complete spontaneous regression. PMID- 11919483 TI - Care of patients with chronic hepatitis C and HIV co-infection: recommendations from the HIV-HCV International Panel. PMID- 11919484 TI - Neutralizing antibodies as a potential secondary protective mechanism during chronic SHIV infection in CD8+ T-cell-depleted macaques. AB - OBJECTIVE: To directly examine the role of CD8+ T cells in controlling viremia and disease during chronic, low-level primate immunodeficiency virus infection in DNA prime/protein boost-vaccinated macaques. BACKGROUND: A cohort of macaques, vaccinated with either a DNA prime/HIV-1 gp160 boost regimen or with gp160 alone was previously protected partially from sequential challenges with non-pathogenic and pathogenic strains of chimeric simian/human immunodeficiency virus (SHIV). In this study, the effect of temporary ablation of CD8+ T cells in these animals was examined. METHODS: Animals were treated with an anti-CD8 antibody and CD8+ T-cell levels in peripheral blood, plasma viral loads, peripheral blood mononuclear cell associated virus levels, neutralizing antibody (nAb) titers and simian immunodeficiency virus Gag-specific CD8+ T-cell numbers were followed. RESULTS: Plasma viremia rose sharply in direct synchrony with a rapid but transient drop in CD8+ T cells. However, although levels of cell-associated virus also rose concomitantly, peak levels were much lower than those in virus-challenged, naive animals. In addition, despite a rise of pathogenic SHIV89.6P RNA levels in three animals, CD4+ T-cell counts remained unchanged. In each of these animals, neutralizing antibody titers against the pathogenic SHIV89.6P strain were high. CONCLUSIONS: The results indicate that CD8+ T cells play a key role in suppressing viremia in a chronically infected host. In addition, the results suggest that in the absence of CD8+ T cells, nAb may act as an effective second line of defense by limiting both the spread of infectious virus to new target cells and CD4+ T-cell loss. PMID- 11919485 TI - Immune reconstitution in HIV-1-infected children on antiretroviral therapy: role of thymic output and viral fitness. AB - OBJECTIVE: To investigate the role of thymic output and viral fitness in immune reconstitution in HIV-1-infected children on antiretroviral therapy. METHODS: Thymic output was studied by measuring levels of T-cell receptor rearrangement excision circles (TREC) in peripheral blood lymphocytes, using a real-time quantitative PCR assay. Recombinant viruses containing pre-therapy or post therapy HIV-1 protease domains were evaluated for viral infectivity in a quantitative single-cycle assay. RESULTS: Eighteen HIV-1-infected children who showed a significant increase in CD4 T-cell count after therapy were studied; HIV 1 plasma viraemia was substantially suppressed in 12 children (virological responders), but not in the other six (virological non-responders). TREC were quantified at baseline, and sequentially during the first 12 months of therapy. Both virological responders and non-responders showed an increase in TREC levels that was inversely correlated with baseline TREC and CD4 T cell counts. Changes in TREC positively correlated with CD4 T-cell count increases in virological responders, but not in non-responders; moreover, the ratios between TREC and CD4 T-cell count increases were higher in non-responders than in responders, suggesting a persistence of peripheral CD4 T-cell loss in the former. Drug resistant viruses with reduced replicative capacity were documented in three out of six non-responders. CONCLUSIONS: These findings indicate that recovery of thymic function is a pivotal event in immune reconstitution, and suggest that CD4 T-cell increase despite persistent viraemia is sustained by a continuous thymic output that compensates peripheral CD4 T-cell depletion which might be slowed down by emerging viruses with reduced fitness. PMID- 11919486 TI - Effect of HIV-1 antiretroviral prophylaxis on hepatic and hematological parameters of African infants. AB - OBJECTIVE: To measure hepatic and hematological parameters among neonates randomized to receive ultra-short antiretroviral regimens. DESIGN: As part of an on-going clinical trial in Malawi, infants born to women who received (early presenters) or did not receive (late presenters) standard intrapartum nevirapine (NVP) dosing were randomized to receive orally either single dose NVP alone or NVP plus zidovudine (twice daily for 1 week). An additional group of untreated infants (born to HIV-uninfected women) was enrolled as a control. METHODS: Laboratory measurements were performed at birth and repeated at 6 weeks of age. Serum alanine aminotransferase (ALT) was measured on approximately 200 infants consecutively enrolled and randomized at the start of the trial. Complete blood count (CBC) was performed on approximately 800 infants at birth and 600 infants at 6 weeks of age. ALT and CBC were also determined on approximately 200 control infants. RESULTS: At birth there were no differences in ALT values between the groups of children. At 6 weeks of age, ALT levels were significantly higher among the treated groups compared with control group (geometric mean of 11.5 U/l for controls and 16.2-19.1 U/l for treated groups; P < 0.0001). Hematological parameters did not differ between groups at birth. At 6 weeks of age, levels of hemoglobin, hematocrit, granulocytes, and platelets were significantly (P < 0.0001) lower among antiviral drug-treated groups compared with controls. These changes were consistent with grade 1 (mild) toxicity, and were more noticeable among HIV-infected infants. CONCLUSIONS: Hepatic and hematologic abnormalities associated with short-term neonatal antiretrovirals among African children are minimal. PMID- 11919487 TI - Indinavir inhibits the glucose transporter isoform Glut4 at physiologic concentrations. AB - OBJECTIVES: To determine the relative sensitivities of glucose transporter isoforms to the protease inhibitor indinavir and to determine the kinetic mechanism of indinavir-mediated Glut4 isoform inhibition. METHODS: The rate of 2 deoxyglucose uptake was measured in Xenopus laevis oocytes heterologously expressing mammalian Glut isoforms. 2-Deoxyglucose uptake was also measured in 3T3-L1 fibroblasts, 3T3-L1 adipocytes, and primary rat adipocytes. RESULTS: The sensitivity to inhibition by indinavir among the Glut isoforms as assayed in the X. laevis oocyte system was as follows in decreasing order: Glut4 >> Glut2 > Glut3 > Glut1 approximately Glut8. 2-Deoxyglucose uptake measurements in insulin stimulated primary rat adipocytes indicated a non-competitive mode of transport inhibition by indinavir under zero-trans conditions with a KI of 15 microM. CONCLUSIONS: Indinavir appears to be a relatively selective inhibitor of the Glut4 isoform. As the concentration required to significantly inhibit insulin stimulated glucose uptake in primary rat adipocytes is well within the physiologic range achieved in therapy, we conclude that direct inhibition of Glut4 contributes to the insulin resistance observed in patients receiving this drug. PMID- 11919488 TI - Acute myeloid leukemia in patients infected with HIV-1. AB - OBJECTIVES: Myelodysplasia is a frequent consequence of HIV infection, but acute myeloid leukemia (AML) is rare. Clinical presentations and outcomes of patients with HIV and subsequent AML are reviewed. METHODS: Five HIV-infected individuals who were subsequently diagnosed with AML were evaluated and treated. A further 42 cases of AML among patients with antecedent HIV infection were identified using MEDLINE, AIDSLINE, and CancerLit searches. RESULTS: HIV infection was present for a median of 48 months (71-180) before AML was diagnosed and the median reported CD4 cell count was 210 x 106 cells/l. In five instances, a delay in diagnosis occurred when cytopenias were initially attributed to HIV or zidovudine-based therapy. In 45 patients, diagnosis was according to the French-American-British (FAB) leukemia classification schema and in two the FAB type was not specified. M2 (n = 15) and M4 (n = 14) subtypes represented 64% (29/45) of reported cases. Patients with a CD4 cell count < 200 x 106 cells/l (n = 11) had a median survival time of 7 weeks, while patients with a CD4 cell count >or= 200 x 106 cells/l (n = 7) had a median survival of 7 months (P = 0.005). Although long-lasting chemotherapy-induced responses were rare, the majority of treated patients did achieve complete hematologic remissions. Treatment-related morbidity did not appear to be excessive. CONCLUSION: In the absence of randomized and prospective clinical studies to guide decision making, this analysis indicates that induction chemotherapy may be a reasonable option for selected HIV-infected patients with AML and adequate immune function. PMID- 11919490 TI - Transplacental passage of protease inhibitors at delivery. AB - OBJECTIVE: Although combinations of different antiretroviral drugs are increasingly used by pregnant HIV-1-infected women, few human data are available to evaluate in utero protease inhibitors (PI) exposure. The aim of this study was to assess the extent of transplacental passage of PI at delivery. METHODS: Pregnant women treated with antiretroviral drugs including PI and/or nevirapine were eligible for the study. Placental transfer was determined by comparison of drug concentrations in blood samples simultaneously collected from a peripheral maternal vein and the umbilical cord at delivery. Drug levels were determined by high-performance liquid chromatography. RESULTS: Thirteen maternal-cord blood sample pairs were evaluable for transplacental passage determination (nine nelfinavir, two ritonavir, one saquinavir, one lopinavir, two nevirapine). Median cord and maternal drug concentrations, respectively, were nelfinavir < 250 and 1110 ng/ml; ritonavir < 250 and 1113 ng/ml; saquinavir < 100 and 350 ng/ml; lopinavir < 250 and 3105 ng/ml and nevirapine 2072 and 2546 ng/ml. The cord-to maternal blood ratio was extremely low for all PI. CONCLUSION: PI do not cross the placenta to an appreciable extent and consequently cannot be expected to exert a direct antiviral activity in utero during the whole dosing interval. Limited transfer may result from their high degree of plasma protein binding and their backwards transport through P-glycoprotein, largely expressed in the placenta. In contrast, nevirapine readily crosses the placental barrier. Such considerations may support treatment decisions in pregnant women. PMID- 11919489 TI - HIV and cytomegalovirus viral load and clinical outcomes in AIDS and cytomegalovirus retinitis patients: Monoclonal Antibody Cytomegalovirus Retinitis Trial. AB - OBJECTIVE: To determine the association of cytomegalovirus (CMV) viremia with mortality and CMV retinitis progression in newly diagnosed and relapsed CMV retinitis. DESIGN: Ancillary study of a randomized, placebo-controlled, phase III clinical trial. PATIENTS: A total of 83 patients with AIDS and CMV retinitis, enrolled during the first phase of the Monoclonal Antibody Cytomegalovirus Retinitis Trial, were administered MSL-109 or placebo as adjuvant therapy for CMV retinitis. MAIN OUTCOME MEASURE(S): Mortality and CMV retinitis progression. RESULTS: Treatment with MSL-109 did not predict either progression of CMV retinitis or mortality. Detection in plasma CMV DNA at baseline predicted mortality, but CMV antigenemia did not. CMV DNA was a better predictor of mortality than a high HIV viral load. Neither CMV DNA nor antigenemia predicted the progression of CMV retinitis. Among newly diagnosed patients, there was a decline in the proportion with detectable CMV viral load and CMV antigenemia in response to anti-CMV therapy. However, there was a rebound in CMV viral load to 25% and CMV antigenemia to 54.6% at 6 months. In relapsed patients, anti-CMV therapy was not associated with a change in the percentage with detectable CMV DNA or CMV antigenemia over time. CONCLUSION: In patients with AIDS and CMV retinitis, the detection of plasma CMV DNA was associated with a higher risk of mortality than was a high HIV viral load. Anti-CMV therapy provided a transient reduction in CMV viremia in newly diagnosed but not relapsed patients with CMV retinitis. Adjuvant therapy with MSL-109 was ineffective in clearing CMV-DNA and CMV antigen from the plasma. PMID- 11919492 TI - Costs of hospital care for HIV-positive and HIV-negative patients at Kenyatta National Hospital, Nairobi, Kenya. AB - OBJECTIVE: To record the costs of hospital care for HIV-positive and -negative patients in Nairobi, and identify costs paid by patients per admission. DESIGN: Cost data were collected on inpatients enrolled in a linked clinical study using standardized costing methods. SETTING: Kenyatta National Hospital, Nairobi's main district hospital. PATIENTS: Consecutive adult medical admissions to one ward over 14 weeks who consented to enrollment; tertiary referrals were excluded. MAIN OUTCOME MEASURE: Average length of stay and cost per patient admission. RESULTS: The hospital costs of 398 patients (163 HIV positive; 33 with clinical AIDS) were analysed. The mean length of stay was 9.3 days and the mean cost per patient admission was US$163. There was no significant difference in costs or mean lengths of stay between HIV-positive and -negative groups, nor were the costs and lengths of stay for clinical AIDS patients significantly different to those for HIV-positive patients without AIDS. The patient charges paid to the hospital per admission, recorded for 344 patients, were on average US$61; and did not differ by HIV status. CONCLUSION: The similar cost patterns for inpatient care irrespective of HIV status or clinical AIDS probably reflects the limited provision of care beyond basic clinical services. Length of stay rather than differing treatment regimes thus appears to be the main cost driver. Private costs of medical care were high and were likely to pressurize households. When resources are limited, the introduction of new, more costly therapies needs careful planning. The study provides cost information for planning care services in resource-poor settings. PMID- 11919491 TI - Selection of drug-resistant HIV-1 mutants in response to repeated structured treatment interruptions. AB - BACKGROUND: A new HIV-1 treatment strategy based on repeated structured treatment interruptions (STI) is currently being evaluated in clinical trials to determine whether immune cell-mediated control of viral replication can be stimulated by intermittent periods of viral replication. The potential for selection of drug resistant quasi-species remains a major concern of such a treatment strategy. METHODS: Plasma and peripheral blood lymphocyte (PBL) samples from 12 patients who had three consecutive STIs were studied. Genotypic analysis was based on population and clonal sequencing. Drug susceptibility and their corresponding replication capacities were evaluated by a single-cycle growth assay. RESULTS: Consistent with a loss of phenotypic susceptibility to lamivudine, the M184V mutation was detected by genotypic analysis (direct and clonal sequencing) in plasma samples collected from two patients at the end of the second or third STI. Longitudinal analysis of patient samples revealed a step-wise increase in the M184V mutation in each patient virus population over successive STIs, despite the lower replicative capacity associated with this mutation in the absence of antiviral agents. CONCLUSION: Drug-resistant virus can rise to high frequencies in chronically HIV-1 infected individuals during consecutive STIs. Evolution of resistance is likely to be more important in patients with prior suboptimal therapies, particularly when few mutations are required for resistance. Maximum care should be taken in designing STI protocols that minimize development of drug resistant mutations that may lead to treatment failure. Thus, drug-resistance testing may be useful before restarting treatment during STI studies. PMID- 11919494 TI - The impact of pregnancy and menopause on CD4 lymphocyte counts in HIV-infected women. AB - OBJECTIVES: To determine indirectly the effect of changes in levels of reproductive hormones on CD4 lymphocyte counts by investigating the impact of pregnancy and menopause on CD4 lymphocyte counts in HIV-infected women. METHODS: Participants were 382 women with a known interval of HIV seroconversion. Review of questionnaires or patient charts provided information on pregnancy and menopause. A linear regression model with a random intercept and slope, which adjusts for multiple CD4 lymphocyte counts per woman, was applied to estimate the CD4 decline following HIV seroconversion and to evaluate the effect of pregnancy and menopause on the CD4 path. RESULTS: The 382 women had a median age of 25 years at seroconversion and yielded 1428 CD4 lymphocyte counts from 3 to 10 years after seroconversion. At 3 years from seroconversion, 20 women had passed the menopause (i.e., the last menses) and five more subsequently passed this point during follow-up; 25 women had a pregnancy after study entry. Postmenopausal women had lower CD4 lymphocyte counts 3 years after seroconversion than premenopausal women (333 vs 399 x 106 cells/l; P = 0.09), and pregnant women had lower counts than non-pregnant women (375 vs 399 x 106 cells/l; P = 0.36). The monthly CD4 decline was not associated with pregnancy and menopause. Adjustment for age did not change the results. CONCLUSIONS: The results suggest that CD4 lymphocyte counts differ between pre- and postmenopausal women, perhaps because of changes in the level of reproductive hormones in the menopause, but associations were not statistically significant. Pregnancy had no statistically significant effect on CD4 lymphocyte counts. PMID- 11919493 TI - Causes and empirical treatment of fever in HIV-infected adult outpatients, Abidjan, Cote d'Ivoire. AB - OBJECTIVES: In Abidjan, HIV prevalence has been estimated at 20% in outpatients attending community clinics. Documenting causes of fever in HIV-infected adult outpatients may help to improve care in these centres with limited facilities. DESIGN: Prospective study. METHODS: We describe all diagnoses and treatments made during febrile episodes in HIV-infected adults participating in the ANRS 059 trial and followed up in a dedicated outpatient centre. RESULTS: Causes of fever could be identified in half of the 269 febrile episodes. Bacterial diseases were the leading identified cause of fever in all CD4 cell count strata (53, 56 and 43% of identified causes in episodes with CD4 count < 200 x 106/l, 200-499 x 106/l, and >or= 500 x 106/l, respectively), followed by malaria (5, 22, and 38%, respectively). Among febrile bacterial diseases, respiratory tract infections and enteritis accounted for 62% of organ involvement, and Streptococcus pneumoniae and non-typhi Salmonella represented 69% of isolated bacterial strains. In these bacterial episodes, an early empirical antibacterial treatment was associated with shorter duration of hospitalization and fever. In the 19 episodes leading to death (7%), the two leading diagnoses were atypical mycobacteriosis (26%) and acute unexplained fever (21%). Death was associated with the absence of antimalarial treatment in the group of acute unexplained fevers. CONCLUSIONS: African HIV treatment guidelines should take into account the predominant role of bacterial infections and malaria in HIV-infected adult outpatients. Reports from other African settings would be useful to compare experiences in algorithms of empirical early antibacterial and antimalarial treatments. PMID- 11919495 TI - HIV protease inhibitors and glucose metabolism. PMID- 11919496 TI - Highly active antiretroviral therapy and cervical intraepithelial neoplasia. AB - Highly active antiretroviral therapy (HAART) has improved HIV prognosis, but its effect on cervical intraepithelial neoplasia (CIN), which is associated with HIV, is uncertain. Among 71 HAART-treated women the prevalence of CIN before HAART was 55%. After a median of 10 months after starting HAART the prevalence had increased to 62% (P = 0.20); 13% of patients experienced regression of a CIN lesion, and this was most strongly associated with a greater increase in CD4 cell count. Such studies will provide the basis for guidelines for monitoring CIN in HIV-positive women in the HAART era. PMID- 11919497 TI - HIV-1 in ethnic Shan migrant workers in northern Thailand. AB - Northern Thailand has one of the highest rates of HIV-1 infection in Southeast Asia. It is also home to a large number of Burmese migrants, believed to be at high risk of HIV. Our 1999 survey of 429 Burmese migrant workers of Shan ethnic origin in Chiang Mai province found a 4.9% rate of HIV-1 prevalence (5.7% men, 3.8% women). This figure is almost double that of comparable population groups in Chiang Mai, e.g. pregnant women and military recruits. HIV prevention programmes are urgently needed for this vulnerable population. PMID- 11919498 TI - Steep increase in HIV prevalence among tuberculosis patients in Ho Chi Minh City. AB - In Vietnam the spread of HIV infection is thought to be limited. In 12 urban districts of Ho Chi Minh City representative samples of tuberculosis patients have undergone HIV testing since 1995. HIV prevalence increased steeply from 0.5% in 1995 to 4% in 2000, with a doubling time of approximately 21 months. This study highlights the need to intensify HIV/AIDS prevention and control in Vietnam. PMID- 11919499 TI - Identification of Pneumocystis carinii DNA in oropharyngeal mouth washes from AIDS children dying of respiratory illnesses. AB - Polymerase chain reaction (PCR) using Pneumocystis carinii-specific primers pAZ 102-H(5'-GTGTACGTTGCAAAGTACTC-3') and pAZ 102-E(5'-GATGGCTGTTTCCAAGCCCA-3') was performed on oropharyngeal washes obtained at autopsy from 22 AIDS children with histologically confirmed P. carinii pneumonia (PCP), and 48 control AIDS children who died from other infections. Fifteen of 22 (68%) PCP samples and none of 48 (0%) control samples had detectable P. carinii DNA (sensitivity 68%; specificity 100%; positive predictive value 100%; negative predictive value 87%). This method requires further validation in clinical practice. PMID- 11919500 TI - Sexual hormones in HIV-infected patients: the influence of antiretroviral therapy. AB - A total of 351 determinations of sexual hormones were carried out in 189 HIV infected men in stable clinical condition. Highly active antiretroviral therapy (HAART) was associated with increased levels of both testosterone and 17beta estradiol, but not with luteinizing hormone (LH) and follicle-stimulating hormone (FSH). Protease inhibitors were more associated with testosterone, and non nucleoside reverse transcriptase inhibitors with 17beta-estradiol. The values of both hormones, but not those of LH and FSH, increased with respect to pre treatment levels in those patients who initiated HAART. PMID- 11919501 TI - Vulnerability of women in an African setting: lessons for mother-to-child HIV transmission prevention programmes. AB - After discussing advantages and risks, only a third of the 290 HIV-infected women included in an intervention study to reduce mother-to-child transmission of HIV in Mombasa, Kenya, informed their partners of their results. Despite careful counselling, 10% subsequently experienced violence or disruption of their relationship. To increase the uptake of interventions to reduce perinatal HIV transmission safely, we recommend the involvement of partners in HIV testing. In addition, the counselling of women has to address methods and skills to deal with violence. PMID- 11919502 TI - Polyphenolic antioxidant (-)-epigallocatechin-3-gallate from green tea as a candidate anti-HIV agent. AB - Epigallocatechin-3-gallate (EGCG), one of the components of green tea, has been suggested to have antiviral activity. To determine the effects of EGCG on HIV infection, peripheral blood lymphocytes were incubated with either LAI/IIIB or Bal HIV strains and increasing concentrations of EGCG. EGCG strongly inhibited the replication of both virus strains as determined by reverse transcriptase and p24 assays on the cell supernatants. PMID- 11919503 TI - Factors associated with persistent high-risk syringe sharing in the presence of an established needle exchange programme. AB - Vancouver has experienced an explosive HIV epidemic despite the presence of a needle exchange programme (NEP). We sought possible explanations for high-risk syringe sharing among Vancouver injection drug users over the period January 1999 to October 2000. Overall, 14% of participants reported high-risk sharing. Although acquiring needles exclusively from the NEP was independently associated with less sharing, we identified several risk factors for persistent sharing, including difficulty accessing sterile needles, bingeing, and frequent cocaine injection. PMID- 11919504 TI - The effectiveness of HIV post-test counselling in determining healthcare-seeking behavior. AB - To examine the relationship between HIV post-test counselling and healthcare seeking behaviors among HIV-positive individuals, we conducted patient interviews and medical chart reviews on 100 HIV-positive patients at a public hospital. When performed, HIV post-test counselling effectively encourages HIV-positive individuals to seek medical care, especially when performed by trained personnel. The consistent use of post-test counselling may improve care for HIV-positive individuals and should be encouraged at all healthcare facilities that conduct HIV testing. PMID- 11919505 TI - Premature vascular lesions in HIV-positive patients: a clockwork bomb that will explode? PMID- 11919506 TI - Carpal tunnel syndrome in HIV patients? PMID- 11919507 TI - Feasibility of antenatal and late HIV testing in pregnant women in Phnom Penh Cambodia: the PERIKAM/ANRS1205 study. PMID- 11919508 TI - Aquaporins in brain: distribution, physiology, and pathophysiology. AB - Water homeostasis in the brain is of central physiologic and clinical importance. Neuronal activity and ion water homeostasis are inextricably coupled. For example, the clearance of K+ from areas of high neuronal activity is associated with a concomitant water flux. Furthermore, cerebral edema, a final common pathway of numerous neurologic diseases, including stroke, may rapidly become life threatening because of the rigid encasement of the brain. A water channel family, the aquaporins, facilitates water flux through the plasma membrane of many cell types. In rodent brain, several recent studies have demonstrated the presence of different types of aquaporins. Aquaporin 1 (AQP1) was detected on epithelial cells in the choroid plexus whereas AQP4, AQP5 and AQP9 were localized on astrocytes and ependymal cells. In rodent brain, AQP4 is present on astrocytic end-feet in contact with brain vessels, and AQP9 is found on astrocytic processes and cell bodies. In basal physiologic conditions, AQP4 and AQP9 appear to be implicated in brain homeostasis and in central plasma osmolarity regulation. Aquaporin 4 may also play a role in pathophysiologic conditions, as shown by the reduced edema formation observed after water intoxication and focal cerebral ischemia in AQP4-knockout mice. Furthermore, pathophysiologic conditions may modulate AQP4 and AQP9 expression. For example, AQP4 and AQP9 were shown to be upregulated after ischemia or after traumatic injuries. Taken together, these recent reports suggest that water homeostasis in the brain is maintained by regulatory processes that, by control of aquaporin expression and distribution, induce and organize water movements. Facilitation of these movements may contribute to the development of edema formation after acute cerebral insults such as ischemia or traumatic injury. PMID- 11919509 TI - Correlation of VEGF and angiopoietin expression with disruption of blood-brain barrier and angiogenesis after focal cerebral ischemia. AB - In an effort to elucidate the molecular mechanisms underlying cerebral vascular alteration after stroke, the authors measured the spatial and temporal profiles of blood-brain barrier (BBB) leakage, angiogenesis, vascular endothelial growth factor (VEGF), associated receptors, and angiopoietins and receptors after embolic stroke in the rat. Two to four hours after onset of ischemia, VEGF mRNA increased, whereas angiopoietin 1 (Ang 1) mRNA decreased. Three-dimensional immunofluorescent analysis revealed spatial coincidence between increases of VEGF immunoreactivity and BBB leakage in the ischemic core. Two to 28 days after the onset of stroke, increased expression of VEGF/VEGF receptors and Ang/Tie2 was detected at the boundary of the ischemic lesion. Concurrently, enlarged and thin walled vessels were detected at the boundary of the ischemic lesion, and these vessels developed into smaller vessels via sprouting and intussusception. Three dimensional quantitative analysis of cerebral vessels at the boundary zone 14 days after ischemia revealed a significant (P < 0.05) increase in numbers of vessels (n = 365) compared with numbers (n = 66) in the homologous tissue of the contralateral hemisphere. Furthermore, capillaries in the penumbra had a significantly smaller diameter (4.8 +/- 2.0 microm) than capillaries (5.4 +/- 1.5 microm) in the homologous regions of the contralateral hemisphere. Together, these data suggest that acute alteration of VEGF and Ang 1 in the ischemic core may mediate BBB leakage, whereas upregulation of VEGF/VEGF receptors and Ang/Tie2 at the boundary zone may regulate neovascularization in ischemic brain. PMID- 11919510 TI - Normobaric hypoxia induces tolerance to focal permanent cerebral ischemia in association with an increased expression of hypoxia-inducible factor-1 and its target genes, erythropoietin and VEGF, in the adult mouse brain. AB - Tolerance to cerebral ischemia is achieved by preconditioning sublethal stresses, such as ischemia or hypoxia, paradigms in which the decrease of O2 availability may constitute an early signal inducing tolerance. In accordance with this concept, this study shows that hypoxia induces tolerance against focal permanent ischemia in adult mice. Normobaric hypoxia (8% O2 of 1-hour, 3-hour, or 6-hour duration), performed 24 hours before ischemia, reduces infarct volume by approximately 30% when compared with controls. To elucidate the mechanisms underlying this neuroprotection, the authors investigated the effects of preconditioning on cerebral expression of hypoxia-inducible factor-1alpha (HIF 1alpha) and its target genes, erythropoietin and vascular endothelial growth factor (VEGF). Hypoxia, whatever its duration (1 hour, 3 hours, 6 hours), rapidly increases the nuclear content of HIF-1alpha as well as the mRNA levels of erythropoietin and VEGF. Furthermore, erythropoietin and VEGF are upregulated at the protein level 24 hours after 6 hours of hypoxia. The authors' findings show that (1) hypoxia elicits a delayed, short-lasting (<72 hours) tolerance to focal permanent ischemia in the adult mouse brain; (2) HIF-1 target genes could contribute to the establishment of tolerance; and (3) this model might be a useful paradigm to further study the mechanisms of ischemic tolerance, to identify new therapeutic targets for stroke. PMID- 11919511 TI - Thrombin-receptor activation and thrombin-induced brain tolerance. AB - The authors previously found that pretreatment with a low dose of thrombin attenuates the brain edema induced by a large dose of thrombin or an intracerebral hemorrhage, and reduces infarct volume after focal cerebral ischemia (i.e., thrombin preconditioning). This study investigated whether thrombin preconditioning is caused by activation of the thrombin receptor, also called protease-activated receptor. In the in vivo studies, thrombin-induced brain tolerance was eliminated by RPPGF (Arg-Pro-Pro-Gly-Phe), a thrombin receptor antagonist. Pretreatment with a thrombin-receptor agonist reduced the amount of edema induced by a large dose of thrombin infused into the ipsilateral basal ganglia 7 days later (81.3 +/- 0.7% vs. 82.6 +/- 0.8% in the control, P < 0.05). In the in vitro study, low doses of thrombin (1 or 2 U/mL) did not induce cell death. However, doses greater than 5 U/mL resulted in dose-dependent lactate dehydrogenase release (P < 0.01). Thrombin and thrombin receptor-activating peptide preconditioning reduced lactate dehydrogenase release induced by a high dose of thrombin (10 and 20 U/mL), whereas RPPGF blocked the effect of thrombin preconditioning in vitro. Western blots indicated that p44/42 mitogen-activated protein kinases were activated after thrombin preconditioning. Finally, inhibition of p44/42 mitogen-activated protein kinases activation by PD98059 abolished the thrombin-preconditioning effect. Results indicate that thrombin induced brain tolerance is in part achieved through activation of the thrombin receptor. Activation of the thrombin receptor in the brain may be neuroprotective. The protective effect of thrombin preconditioning is achieved through the p44/42 mitogen-activated protein kinase signal-transduction pathway. PMID- 11919512 TI - Three steps of neural stem cells development in gerbil dentate gyrus after transient ischemia. AB - The stage of neurogenesis can be divided into three steps: proliferation, migration, and differentiation. To elucidate detailed relations between these three steps after ischemia, the authors evaluated the three steps in the adult gerbil dentate gyrus (DG) after 5 minutes of transient global ischemia using bromodeoxyuridine (BrdU), highly polysialylated neural cell adhesion molecule (PSA-NCAM), and neuronal nuclear antigen (NeuN) and glial fibrillary acidic protein (GFAP) as markers for proliferation, migration, and differentiation, respectively. Bromodeoxyuridine-labeled cells increased approximately sevenfold, and PSA-NCAM-positive cells increased approximately threefold in the subgranular zone (SGZ) with a peak 10 days after ischemia. Bromodeoxyuridine-labeled cells with PSA-NCAM expression were first detected both in the SGZ and the granule cell layer (GCL) 20 days after ischemia and gradually decreased after that, whereas BrdU-labeled cells with NeuN gradually increased in the GCL until 60 days after ischemia. A few BrdU-labeled cells with GFAP expression were detected in DG after ischemia; no PSA-NCAM-positive cells with GFAP expression were detected, but the radial processes of glial cells were partly in contact with PSA-NCAM-positive cell bodies and dendrites. These results suggest that neural stem cell proliferation begins at the SGZ, and that the cells then migrate into the GCL and differentiate mainly into neuronal cells. The majority of these three steps finished in 2 months after transient global ischemia. PMID- 11919513 TI - Role of caspase-3 activation in cerebral ischemia-induced neurodegeneration in adult and neonatal brain. AB - These studies have addressed the role of caspase-3 activation in neuronal death after cerebral ischemia in different animal models. The authors were unable to show activation of procaspase-3 measured as an induction of DEVDase (Asp-Glu-Val Asp) activity after focal or transient forebrain ischemia in rats. DEVDase activity could not be induced in the cytosolic fraction of the brain tissue obtained from these animals by exogenous cytochrome c/dATP and Ca2+. However, the addition of granzyme B to these cytosolic fractions resulted in a significant activation of DEVDase, confirming that the conditions were permissive to analyze proteolytic cleavage of the DEVD-AMC (7-amino-4-methyl-coumarin) substrate. Consistent with these findings, zVal-Ala-Asp-fluoromethylketone administered after focal ischemia did not have a neuroprotective effect. In contrast to these findings, a large increase in DEVDase activity was detected in a model of hypoxic ischemia in postnatal-day-7 rats. Furthermore, in postnatal-day-7 animals treated with MK-801, in which it has been suggested that excessive apoptosis is induced, the authors were unable to detect activation of DEVDase activity but were able to induce it in vitro by the addition of cytochrome c/dATP and Ca2+ to the cytosolic fraction. Analysis of cytochrome c distribution did not provide definitive evidence for selective cytochrome c release in the permanent focal ischemia model, whereas in the transient model a small but consistent amount of cytochrome c was found in the cytosolic fraction. However, in both models the majority of cytochrome c remained associated with the mitochondrial fraction. In conclusion, the authors were unable to substantiate a role of mitochondrially derived cytochrome c and procaspase-3 activation in ischemia-induced cell death in adult brain, but did see a clear induction of caspase-3 in neonatal hypoxia. PMID- 11919514 TI - Activation of mitochondrial ATP-dependent potassium channels protects neurons against ischemia-induced death by a mechanism involving suppression of Bax translocation and cytochrome c release. AB - Neurons express a variety of plasma-membrane potassium channels that play important roles in regulating neuronal excitability and synaptic transmission, but also contain mitochondrial ATP-sensitive potassium channels, the functions of which are unknown. Studies of cardiac cells suggest that similar mitochondrial ATP-sensitive potassium channels are involved in the process of ischemic preconditioning, suggesting a role in regulating cell survival. The authors report that mice given diazoxide, an activator of mitochondrial ATP-sensitive potassium channels, exhibited a large (60% to 70%) decrease in cortical infarct size after permanent occlusion of the middle cerebral artery. Diazoxide decreases neuronal apoptosis and increases astrocyte survival and activation in the penumbral region of the ischemic cortex. The neuroprotective effect of diazoxide is abolished by 5-hydroxydecanoate, a selective antagonist of mitochondrial ATP sensitive potassium channels. Studies of cultured hippocampal neurons reveal that diazoxide depolarizes mitochondria, prevents cytochrome c release, and protects cells against death induced by staurosporine and chemical hypoxia. Diazoxide increased the levels of Bcl2 and inhibited the association of Bax with mitochondria in neurons exposed to an apoptotic insult, suggesting that activation of mitochondrial ATP-sensitive potassium channels may stabilize mitochondrial function by differentially modulating proapoptotic and antiapoptotic proteins. Collectively, the data suggest that mitochondrial ATP sensitive potassium channels play a key role in modulating neuronal survival under ischemic conditions, and identify agents that activate mitochondrial ATP sensitive potassium channels as potential therapeutics for stroke and related neurodegenerative conditions. PMID- 11919515 TI - Mitogen-activated protein kinase inhibition in traumatic brain injury: in vitro and in vivo effects. AB - The authors provide the first in vitro and in vivo evidence that perturbations in mitogen-activated protein kinase (MAPK) signal-transduction pathways are involved in the pathophysiology of traumatic brain injury. In primary rat cortical cultures, mechanical trauma induced a rapid and selective phosphorylation of the extracellular signal-regulated kinase (ERK) and p38 kinase, whereas there was no detectable change in the c-jun N-terminal kinase (JNK) pathway. Treatment with PD98059, which inhibits MAPK/ERK 1/2, the upstream activator of ERK, significantly increased cell survival in vitro. The p38 kinase and JNK inhibitor SB203580 had no protective effect. Similar results were obtained in vivo using a controlled cortical impact model of traumatic injury in mouse brain. Rapid and selective upregulation occurred in ERK and p38 pathways with no detectable changes in JNK. Confocal immunohistochemistry showed that phospho-ERK colocalized with the neuronal nuclei marker but not the astrocytic marker glial fibrillary acidic protein. Inhibition of the ERK pathway with PD98059 resulted in a significant reduction of cortical lesion volumes 7 days after trauma. The p38 kinase and JNK inhibitor SB203580 had no detectable beneficial effect. These data indicate that critical perturbations in MAPK pathways mediate cerebral damage after acute injury, and further suggest that ERK is a novel therapeutic target in traumatic brain injury. PMID- 11919516 TI - Ischemia-induced brain damage depends on specific gap-junctional coupling. AB - Ischemic brain injury results in neuronal loss and associated neurologic deficits. Although there is some evidence that intercellular communication via gap junctions can spread oxidative cell injury, the possible role of gap junctional communication in ischemia-induced cell death is the object of debate. Because gap junctions directly connect the cytoplasms of coupled cells, they offer a way to propagate stress signals from cell to cell. The authors investigated the contribution of gap-junctional communication to cell death using an in vitro ischemia model, which was reproduced by submersion of organotypic hippocampal slices into glucose-free deoxygenated medium. The gap-junctional blocker carbenoxolone significantly decreased the spread of cell death, as measured by propidium iodide staining, over a 48-hour period after the ischemic episode. Carbenoxolone ameliorated the hypoxia-induced impairment of the intrinsic neuronal electrophysiologic characteristics, as measured by whole-cell patch clamp recordings. To determine whether specific connexins were involved in the spread of postischemic cell death, the authors partially reduced the synthesis of specific connexins using antisense oligodeoxynucleotides. Simultaneous knockdown of two connexins localized mostly in neurons, connexins 32 and 26, resulted in significant neuroprotection 48 hours after the hypoxic hypoglycemic episode. Similarly, partial reduction of the predominant glial connexin 43 significantly decreased cell death. These results indicate that gap junctional communication contributes to the propagation of hypoxic injury and that specific gap junctions could be a novel target to reduce brain damage. PMID- 11919517 TI - Neuroprotection by a bile acid in an acute stroke model in the rat. AB - Tauroursodeoxycholic acid (TUDCA), a hydrophilic bile acid, is a strong modulator of apoptosis in both hepatic and nonhepatic cells, and appears to function by inhibiting mitochondrial membrane perturbation. Excitotoxicity, metabolic compromise, and oxidative stress are major determinants of cell death after brain ischemia-reperfusion injury. However, some neurons undergo delayed cell death that is characteristic of apoptosis. Therefore, the authors examined whether TUDCA could reduce the injury associated with acute stroke in a well characterized model of transient focal cerebral ischemia. Their model of middle cerebral artery occlusion resulted in marked cell death with prominent terminal deoxynucleotidyl transferase-mediated 2;-deoxyuridine 5;-triphosphate-biotin nick end labeling (TUNEL) within the ischemic penumbra, mitochondrial swelling, and caspase activation. Tauroursodeoxycholic acid administered 1 hour after ischemia resulted in significantly increased bile acid levels in the brain, improved neurologic function, and an approximately 50% reduction in infarct size 2 and 7 days after reperfusion. In addition, TUDCA significantly reduced the number of TUNEL-positive brain cells, mitochondrial swelling, and partially inhibited caspase-3 processing and substrate cleavage. These findings suggest that the mechanism for in vivo neuroprotection by TUDCA is, in part, mediated by inhibition of mitochondrial perturbation and subsequent caspase activation leading to apoptotic cell death. Thus, TUDCA, a clinically safe molecule, may be useful in the treatment of stroke and possibly other apoptosis-associated acute and chronic injuries to the brain. PMID- 11919519 TI - Calcium compartments in brain. AB - Excellent progress has been made toward understanding the physiology and pharmacology of specific calcium-related cellular processes of the brain, but few studies have provided an integrated view of brain calcium kinetics. To further the knowledge of the size and binding properties of brain calcium compartments, the authors have conducted a series of experiments in hippocampal brain slices exposed to high and low extracellular calcium. Slices were incubated in buffers containing 0.001 to 4.5 mmol/L calcium for up to 75 minutes. Slice calcium content was analyzed by three methods: exchange equilibrium with 45Ca, synchrotron-radiation-induced x-ray emission, and inductively coupled plasma. Data were analyzed using a model based on a Langmuir isotherm for two independent sites, with additional extracellular and bound compartments. In parallel experiments, altered low calcium had no effect on slice histology and only mild effects on slice adenylates. When combined with prior 45Ca and fluorescent probe binding experiments, these results suggest that there are at least five kinetically distinct calcium compartments: (1) free extracellular (approximately 10%); (2) loosely associated extracellular plasma membrane (approximately 55%); (3) intracellular compartment with moderate avidity (approximately 17%); (4) tightly bound, nonexchangeable intracellular compartment ( approximately 15%); and (5) free cytoplasmic (<0.01%). If only the third compartment is considered a potential calcium buffer, then the buffering ratio is calculated to be approximately 2,700:1, but if the second compartment is also included, then the buffering ratio would be approximately 13,000:1. This may explain the wide range of estimates observed by fluorescent probe studies. PMID- 11919518 TI - Oxidation of bilirubin produces compounds that cause prolonged vasospasm of rat cerebral vessels: a contributor to subarachnoid hemorrhage-induced vasospasm. AB - The authors have previously shown that bilirubin-oxidation products (BOXes) are present in CSF of subarachnoid hemorrhage patients with vasospasm, and that BOXes cause vasoconstriction in vitro. This study determined whether BOXes cause vasospasm in vivo. Identical volumes of either lysed blood or standardized amounts of BOXes were injected into the cisterna magna of adult rats. BOX injections caused 6 of 10 rats to die within 10 minutes, whereas 12 of 12 rats survived for 24 hours after blood injections. The mechanism for this significant (P < or = 0.01) increase in mortality was unclear. To directly test whether BOXes produced vasospasm, a cranial window technique was used. Application of 20 microL of 10-micromol/L bilirubin had little effect on the vessels. However, application of BOXes produced marked, dose-dependent small artery and arteriole vasospasm that approached a 90% decrease in diameter by 40 minutes after application in some vessels, and persisted for at least 24 hours. To determine if BOX-mediated vasospasm led to cortical injury, histology and immunocytochemistry were performed on animals that survived for 24 hours. There was a BOX-related stress protein response for HSP25 and HSP32 (HO-1) without evidence of infarction. The finding that the BOXes produce vasospasm of cerebral vessels in vivo, in conjunction with BOXes being found in CSF of vasospasm patients, supports our hypothesis that BOXes contribute to or cause cerebral vasospasm after subarachnoid hemorrhage. PMID- 11919520 TI - Quantitative analysis for estimating binding potential of the brain serotonin transporter with [11 C]McN5652. AB - SUMMARY: [11C](+)McN5652 is a selective serotonin reuptake inhibitor with subnanomolar potency for the serotonin transporter, and is currently being used for positron emission tomography studies. However, quantification of the regional [11C](+)McN5652 binding potential in vivo is a controversial issue because of its complex characteristics. The authors examined the regional differences in nonspecific binding and proposed simple methods for estimating the binding potential of [11C](+)McN5652. The regional difference in nonspecific binding was evaluated by the activity ratio of the thalamus compared with the cerebellum for inactive-isomer [11C](-)McN5652 and [11C](+)McN5652 saturation studies. The distribution volume of the thalamus was approximately 1.16 times larger than that of the cerebellum. The thalamus-to-cerebellum distribution volume ratio was estimated by nonlinear least square and graphical methods, with and without arterial input function. The graphical method with k2; without blood sampling was practical and most applicable for estimation of the distribution volume ratio because this method is more stable than the nonlinear least square method in the simulation study. Binding potential estimated with the distribution volume ratio of [11C](+)McN5652 and the correction with distribution volume ratio of [11C]( )McN5652 represent the most reliable parameters for the assessment of serotonin transporter binding. PMID- 11919521 TI - Low-dose Carbicarb improves cerebral outcome after asphyxial cardiac arrest in rats. AB - STUDY OBJECTIVE: Controversy surrounds the use of buffers during cardiac arrest to correct acidosis. The objective of this study was to determine whether attenuation or neutralization of cerebral acidosis by Carbicarb alters hippocampal glutamate levels, neuronal cell death, and neurologic deficits after reperfusion from asphyxial cardiac arrest in rats. METHODS: Rats were prospectively randomized to either a control (n=45), low-dose Carbicarb (LDC; 3 mL/kg, n=45), or high-dose Carbicarb (HDC; 6 mL/kg, n=45) group in a blinded fashion during resuscitation after 8 minutes of asphyxial cardiac arrest. Microdialysis was used to assess brain pH and glutamate. A neurologic deficit score and neuronal cell death in the hippocampus were determined at day 7. RESULTS: Resuscitation was greatest in LDC rats (42/45) and least in HDC rats (28/45) versus that in control rats (34/45). Brain pH was higher in the LDC and HDC rats 10 minutes after resuscitation and remained higher than that of control rats for 120 minutes after resuscitation. Glutamate levels at 10 to 120 minutes after reperfusion were lowest in the LDC rats. LDC rats had the lowest neurologic deficit score (1+/-2) versus that of control rats (13+/-8) and HDC rats (19+/-6). Hippocampal neuronal cell death was lowest in LDC rats (30+/-20) versus that in control rats (86+/-47) and HDC rats (233+/-85). CONCLUSION: LDC administered during resuscitation from asphyxial cardiac arrest attenuated acidosis, improved resuscitation, and reduced neurologic deficits and the number of dead hippocampal neurons. Neutralization of cerebral acidosis with HDC increased the number of dead hippocampal neurons and neurologic deficits after resuscitation from cardiac arrest in rats. PMID- 11919522 TI - A neural computational aid to the diagnosis of acute myocardial infarction. AB - STUDY OBJECTIVE: Accurate identification of the presence of acute myocardial infarction in adult patients who present to the emergency department with anterior chest pain remains elusive. The artificial neural network is a powerful nonlinear statistical paradigm for the recognition of complex patterns, with the ability to maintain accuracy when some data required for network function are missing. Earlier studies revealed that the artificial neural network is able to accurately identify acute myocardial infarction in patients experiencing chest pain. However, these studies did not measure network performance in real time, when a significant amount of data required for network function may not be available. They also did not use chemical cardiac marker data. METHODS: Two thousand two hundred four adult patients presenting to the ED with anterior chest pain were used to train an artificial neural network to recognize the presence of acute myocardial infarction. Only data available at the time of initial patient evaluation were used to replicate the conditions of real-time patient evaluation. Forty variables from patient histories, physical examinations, ECG results, and chemical cardiac marker determinations were used to train and then test the network. RESULTS: The network correctly identified 121 of the 128 patients (sensitivity 94.5%; 95% confidence interval 90.6% to 97.9%) with myocardial infarction at a specificity of 95.9% (95% confidence interval 93.0% to 98.5%), despite the fact that an average of 5% (individual range 0% to 35%) of the input data required by the network were missing on all patients. CONCLUSION: Network accuracy and the maintenance of that accuracy when some data required for function are unavailable suggest that the artificial neural network may be a potential real time aid to the diagnosis of acute myocardial infarction during initial patient evaluation. PMID- 11919523 TI - An emergency department observation unit protocol for acute-onset atrial fibrillation is feasible. AB - STUDY OBJECTIVE: We sought to describe the feasibility of an emergency department observation unit (EDOU) treatment protocol for the management of uncomplicated acute-onset atrial fibrillation (AAF). METHODS: This descriptive case series took place at a major suburban, university-affiliated teaching hospital. Patients were prospectively enrolled in an EDOU treatment protocol if they had uncomplicated AAF that failed initial ED attempts to convert to sinus rhythm. In the EDOU, patients underwent ECG monitoring, serial creatine kianse MB measurements, and further rate control with optional electrical cardioversion. Primary outcomes measured were EDOU rate of conversion to sinus rhythm, rate of discharge home, length of stay, positive diagnostic outcomes, complications of AAF, and 7-day return visits. RESULTS: Sixty-seven patients were studied. Patients were symptomatic for a median of 4.0 hours, had mean initial ED pulse rates of 137+/ 23 beats/min, and spent 4.7+/-2.2 hours in the ED before transfer to the EDOU. While in the EDOU, 55 (82%) patients converted to sinus rhythm. Five (7%) patients were admitted because of positive test results: 2 for myocardial infarction, 2 for fever, and 1 for ventricular tachycardia. Twelve (18%) patients remained in atrial fibrillation, with 9 admitted and 3 discharged. Overall, 81% of patients were discharged in 11.8+/-7.0 hours, and 19% were admitted after 17.6+/-9.5 hours of observation. Three discharged patients returned within 7 days, 2 for uncomplicated recurrent AAF and 1 for chest pain subsequently found to be noncardiac in origin. There were no major complications attributable to the EDOU protocol. CONCLUSION: Selected patients with AAF for whom initial ED management fails can subsequently be managed in an EDOU with a high short-term conversion and discharge rate. PMID- 11919524 TI - Subclassification of indeterminate pelvic ultrasonography: prospective evaluation of the risk of ectopic pregnancy. AB - STUDY OBJECTIVE: We sought to determine the frequency of ectopic pregnancy among subclasses of indeterminate ultrasonographic examinations. METHODS: A prospective observational study was performed from January 1, 1995, to August 31, 2000, on consecutive emergency department patients in the first trimester of pregnancy with a chief complaint of abdominal pain or vaginal bleeding and who had an indeterminate transvaginal ultrasonographic examination at the time of the ED visit. Patients were excluded if lost to follow-up. Ultrasonographic examinations were subclassified into 5 groups (ie, empty uterus, nonspecific fluid, echogenic material, abnormal sac, normal sac) on the basis of a previously published classification system. Patients were followed up until the diagnosis of ectopic pregnancy was either confirmed or excluded. The frequencies of ectopic pregnancy, along with 95% confidence intervals (CIs), were calculated for each of the subclasses. The relative risk of ectopic pregnancy was calculated when appropriate. RESULTS: Seven hundred eighty patients with indeterminate ultrasonographic examinations were identified. One hundred forty-five were lost to follow-up, and therefore, 635 were enrolled. The frequency of ectopic pregnancy for each subclass is as follows: empty uterus, 36 of 259 (13.9%; 95% CI 10.1% to 18.5%); nonspecific fluid, 6 of 127 (4.7%; 95% CI 1.9% to 9.6%); echogenic material, 4 of 93 (4.3%; 95% CI 1.4% to 10.5%); abnormal sac, 0 of 103 (0%; 95% CI 0.0% to 2.9%); and normal sac, 0 of 53 (0%; 95% CI 0.0% to 5.5%). The relative risk of ectopic pregnancy in patients with an empty uterus versus in those without an empty uterus was 5.2 (95% CI 2.6 to 10.2). CONCLUSION: In our sample, patients with an empty uterus at ultrasonography had the highest frequency of ectopic pregnancy, with a relative risk of ectopic pregnancy 5 times greater than that of the other 4 subclasses. PMID- 11919525 TI - Trends in the use and capacity of California's emergency departments, 1990-1999. AB - STUDY OBJECTIVE: Concerns over the ability of the nation's emergency departments to meet current demands are growing among the public and health care professionals. Data supporting perceptions of inadequate capacity are sparse and conflicting. We describe changes in the use and capacity of California's EDs between 1990 and 1999, as well as trends in severity of patient illness or injury. METHODS: Data from California's Office of Statewide Health Planning and Development (OSHPD), which describe all hospital and health service use in the state, were analyzed and later verified using a telephone survey of all 320 open EDs in California. Six variables were analyzed: hospital's ownership type (public or private), total number of annual ED visits, severity of patient illness or injury (percentage of visits categorized as critical, urgent, or nonurgent), number of ED beds, proximity to a closed ED, and teaching status. We tested 2 main hypotheses: (1) Have statewide ED visits, ED beds, visits per ED, and visits per bed increased or decreased between 1990 and 1999? and (2) Has severity of patient illness or injury, as reported to OSHPD, changed over the past decade? State level data were analyzed using ordinary least-squares regression. Hospital level data were analyzed using repeated measures analyses. RESULTS: The number of EDs in California decreased by 12% (P <.0001). The number of ED treatment stations (ie, physical spaces for the treatment of patients) increased by 687 (16%) statewide (P =.0001), or an average of 79 beds per year. The average annual change in ED visits was not statistically significant (P =.5), whereas visits per ED increased by 27% for all EDs (P <.0001), although with differing trends noted at public and private hospitals. At private hospitals, the average increase was 512 visits/ED each year, whereas at public hospitals, visits decreased by an average of 1,085 visits/ED each year (P <.0001). Overall, critical visits per ED increased by 59% (P <.0001), and nonurgent visits per ED decreased by 8% (P <.0001). CONCLUSION: The number of EDs in California decreased significantly during the 1990s, whereas the number of ED beds increased. Increases in visits per ED, beds per ED, and in the proportion of patients categorized as critical may help explain the perception that ED capacity is inadequate to meet growing demand. PMID- 11919527 TI - Practice guidelines and performance measures in emergency medical services for children. AB - Practice guidelines and performance measures are critical elements of an effective quality improvement process for emergency medical services for children (EMSC). Practice guidelines address the clinical management of individual patients, and performance measures assess the quality of care delivered to a population. The public and private sectors have invested considerable resources in developing practice guidelines and performance measures to improve the quality of health care services. As organizations continue development efforts, health care professionals who are actively involved in emergency care must collaborate to develop guidelines that address the unique physiologic, psychologic, and cultural needs of children. The Emergency Medical Services for Children Managed Care Task Force recommended the development of a series of white papers to focus on issues related to practice guidelines and performance measures in EMSC. The Maternal and Child Health Bureau, Health Resources and Services Administration, the National Highway Traffic Safety Administration, and the Robert Wood Johnson Foundation jointly sponsored the project. The paper was developed by a panel selected from a pool of experts in managed care, quality improvement, and emergency medical services. After a review of the literature, the panelists met to discuss critical issues related to practice guidelines and performance measures in EMSC. The panelists developed recommendations that can serve as resources for managed care organizations, health care providers, professional associations, and governmental policy makers. The panel recognized the lack of nationally recognized pediatric emergency care guidelines and performance measures and called for immediate action in these areas. PMID- 11919526 TI - A randomized clinical trial comparing oral ondansetron with placebo in children with vomiting from acute gastroenteritis. AB - STUDY OBJECTIVE: Vomiting in children suffering from acute gastroenteritis interferes with the oral rehydration process and equally frustrates parents and health care providers. Adjuncts such as promethazine and metoclopramide are less than optimally effective and are associated with side effects. Ondansetron, a 5 HT3 receptor antagonist marketed as Zofran, is a safe and effective antiemetic used extensively in oncology and postoperative patients. We evaluate the effect of the antiemetic ondansetron versus placebo on the clinical outcome of patients with vomiting from gastroenteritis in a pediatric emergency department. METHODS: This was a randomized, prospective, double-blind clinical trial in a university affiliated children's hospital ED. Children between the ages of 6 months and 12 years who had vomited at least 5 times during the preceding 24 hours were randomized to receive either oral ondansetron or a taste- and color-matched placebo. Oral rehydration was commenced 15 minutes later at 5 mL/min per standard oral rehydration protocols. Patients were discharged after they voided and continued standard oral rehydration at home with the introduction of a bananas, rice, applesauce, and toast (BRAT) diet after the first 24 hours. Any patient requiring admission was considered a treatment failure, and no further doses were given. Discharged patients were given 5 additional doses to be used every 8 hours, and they were contacted by telephone 24 and 48 hours after discharge to record the number of episodes of vomiting and diarrhea. The parents were also required to complete a diary of the same information, which was mailed to the investigators for confirmation of the telephone data. RESULTS: One hundred forty five patients were enrolled, of whom 51% (n=74) were randomized to ondansetron. At baseline, age distribution, sex, and severity of illness did not differ between the ondansetron and placebo groups. During the observation period in the ED, the median number of episodes of vomiting was 0 in both groups, but the rank sum of vomiting episodes was significantly lower in the ondansetron group (P =.001). The number of episodes of emesis in the ED after enrollment ranged from 0 to 7 in the placebo group and 0 to 2 in the ondansetron group. During the 48 hours of follow-up, the median number of episodes of vomiting remained 0, with no statistically significant difference between the groups. There was no statistically significant difference in the rank sum of episodes of diarrhea in the ED between the groups (P =.622); however, during the next 48 hours, the patients in the ondansetron group had significantly more diarrhea than the placebo group. A lower proportion of patients receiving ondansetron compared with placebo required intravenous fluid therapy (P =.015). The admission rate was also lower in patients receiving ondansetron (P =.007). The revisit rate was higher in the ondansetron group compared with the placebo group (P =.047). CONCLUSION: Ondansetron was effective in reducing the emesis from gastroenteritis during the ED phase of oral rehydration and in lowering the rates of intravenous fluid administration and hospital admission. PMID- 11919528 TI - Diagnosis and management of acute otitis media in the urgent care setting. AB - The prevalence of otitis media is increasing, which affects health care resource utilization across all segments, including the urgent care setting. One of the greatest challenges in the management of acute otitis media (AOM) is the effective treatment of cases caused by pathogens that are resistant to commonly used antibiotics. Whereas the production of beta-lactamases among strains of Haemophilus influenzae and Moraxella catarrhalis is an important consideration for antimicrobial therapy, the high prevalence of resistance to penicillin and other classes of antibiotics among strains of Streptococcus pneumoniae represents a greater clinical concern. The Centers for Disease Control and Prevention (CDC) recently convened the Drug Resistant S. pneumoniae Therapeutic Working Group to develop evidence-based recommendations for the treatment of AOM in an era of prevalent resistance. The recommendations from this group included amoxicillin as the preferred first-line drug because of the demonstrated activity against penicillin-intermediate and -resistant strains of S. pneumoniae, using higher dosages of up to 90 mg/kg per day in certain settings. For patients in whom initial treatment is unsuccessful after 3 days, the recommended agents included high-dose amoxicillin-clavulanate (for activity against beta-lactamase-producing pathogens), clindamycin, cefuroxime axetil, or 1 to 3 doses of intramuscular ceftriaxone. The principles set forth in these guidelines can assist the therapeutic decisionmaking process for practitioners in the urgent care setting. PMID- 11919529 TI - The Frontlines of Medicine Project: a proposal for the standardized communication of emergency department data for public health uses including syndromic surveillance for biological and chemical terrorism. AB - The Frontlines of Medicine Project is a collaborative effort of emergency medicine (including emergency medical services and clinical toxicology), public health, emergency government, law enforcement, and informatics. This collaboration proposes to develop a nonproprietary, "open systems" approach for reporting emergency department patient data. The common element is a standard approach to sending messages from individual EDs to regional oversight entities that could then analyze the data received. ED encounter data could be used for various public health initiatives, including syndromic surveillance for chemical and biological terrorism. The interlinking of these regional systems could also permit public health surveillance at a national level based on ED patient encounter data. Advancements in the Internet and Web-based technologies could allow the deployment of these standardized tools in a rapid time frame. PMID- 11919531 TI - Procedural sedation terminology: moving beyond "conscious sedation". PMID- 11919530 TI - Overcrowding in emergency departments: increased demand and decreased capacity. PMID- 11919532 TI - How to find evidence when you need it, part 2: a clinician's guide to MEDLINE: the basics. PMID- 11919533 TI - Episodic stridor with latex nipple use in a 2-month-old infant. AB - Latex allergy in the pediatric population is most commonly identified in patients who have undergone multiple operations for neural tube defects or exstrophic genitourinary anomalies. However, there are a significant number of children who, without the usual risk factors, clinically and/or serologically appear to be latex allergic. There is sporadic information in the medical literature regarding reactions to latex allergens in household items, especially in patients younger than 1 year old. Several recent reports even support the existence of reactions to latex pacifiers. We report a case of an atopic 2-month-old infant who experienced the previously unreported reaction of repeated stridor on exposure to a latex nipple while feeding. It is important that clinicians recognize stridor as a potential reaction to latex in infants. PMID- 11919534 TI - Application of chronobiology to resident physician work scheduling. PMID- 11919535 TI - Guide for preparing medical directors. PMID- 11919536 TI - Commentary: medical direction for out-of-hospital emergency medical services. PMID- 11919539 TI - Demographic analysis and needs assessment of rural emergency departments in New Mexico (DANARED-NM). PMID- 11919541 TI - Remembering September 11th. PMID- 11919540 TI - Indications for appropriate platelet glycoprotein IIb/IIIa inhibitor therapy. PMID- 11919542 TI - Pastoral care as adjuvant therapy. PMID- 11919544 TI - Leptin and the neurobiology of eating disorders. PMID- 11919545 TI - Leptin in anorexia nervosa and bulimia nervosa: importance of assay technique and method of interpretation. AB - Studies of the role of leptin in patients with anorexia nervosa and bulimia nervosa have conflicted in their data and interpretation. Such differences may be a result of the assay methods used or the way results are compared with those from normal controls. To investigate these possibilities, we analyzed serum leptin levels in anorexic, bulimic, obese, and control individuals, thereby spanning the full range of human body weights, using three frequently employed commercial kits. Kits from Linco (St Louis, MO) and DSL (Webster, TX) employ a radioimmunoassay method, and the R&D Systems kit (Minneapolis, MN) uses an enzyme linked immunosorbent assay. We found that the three kits provide results that are highly linearly correlated with each other and remarkably linearly related to percent ideal body weight (%IBW) over more than three orders of magnitude (Linco, r = 0.90; R&D, r = 0.87; DSL, r = 0.86). For very low leptin levels, the more sensitive kits from R&D and Linco appeared to give more reliable results. Measurement method does not appear to explain the literature conflicts. We found that patients with anorexia nervosa have serum leptin values that lie above the line extrapolated from the %IBW/leptin curve generated from analysis of all non anorexic patients. Therefore, in anorexia nervosa, inappropriately high leptin levels for %IBW may contribute to a blunted physiologic response to underweight and consequent resistance to dietary treatment. By contrast, most bulimic patients have leptin levels significantly below those predicted from the same %IBW/leptin curve. The relative leptin deficiency in bulimic subjects may contribute to food-craving behavior. We propose that using the %IBW/ leptin curve can facilitate identification of true pathophysiologic abnormalities in eating disordered individuals and provide a basis for the design of therapeutic interventions or monitoring of response to treatment. PMID- 11919546 TI - Eicosanoids in sickle cell disease: potential relevance of neutrophil leukotriene B4 to disease pathophysiology. AB - Neutrophil activation with the release of intracellular granule contents has been observed in sickle cell disease (SCD). Because leukotriene B(4) (LTB(4)), a 5 lipoxygenase metabolite of arachidonic acid in neutrophils, is a chemoattractant and enhances neutrophil adhesion to endothelium, we assessed plasma levels of this metabolite in controls (n = 9) and individuals with SCD, SS genotype, both in basal "steady state" (n = 37) and during episodes of vaso-occlusion (n = 10) and acute chest syndrome (n = 5). Thirteen patients with SCD, SC genotype, in steady state were also studied. Although no significant differences were noted between the control (136 +/- 32 fmol/mL) and SC genotype (177 +/- 83 fmol/mL, P >.15), LTB(4) levels were markedly increased in patients with SS genotype in basal steady state (207 +/- 64 fmol/mL, P <.003) compared with those in controls. Values were further increased during vaso-occlusion (264 +/- 94 fmol/mL) and acute chest syndrome (363 +/- 124 fmol/mL). These levels were significantly different from measurements taken during steady state (P <.04 and P <.0001, respectively). No correlation was noted between LTB(4) level and total white cell or neutrophil count. Additionally, the significant correlation noted in SCD between increased levels of plasma LTB(4) and soluble L-selectin (P <.03) reflects neutrophil activation. We also observed an effect of LTB(4) on red cell endothelial adhesion at concentrations that appear clinically relevant (1-10 pmol/mL) with concomitant up-regulation of mRNA for the endothelial vitronectin receptor. These properties of LTB(4) are relevant to disease pathophysiology, providing further evidence of the contribution of the neutrophil to the proinflammatory and proadhesive phenotype in SCD. PMID- 11919547 TI - Interleukin-4 inhibits tumor necrosis factor-alpha-induced and spontaneous apoptosis of biomaterial-adherent macrophages. AB - Biocompatibility of implanted materials is determined by the host foreign-body response, which is comprised of cellular (adherent monocytes and macrophages) and soluble (secreted cytokines) components. Modulating the presence, activity or both of adherent macrophages may increase or decrease the biocompatibility of implants because these cells remain adherent to the implant surface and fuse to form foreign-body giant cells (FBGCs), leading to failure of the implant. An attractive mechanism of eliminating these cells is through the induction of apoptosis; therefore ways of inducing or inhibiting apoptosis of biomaterial adherent inflammatory cells are being investigated. We hypothesized that interleukin-4 (IL-4) promotes macrophage survival by inhibiting tumor necrosis factor-alpha (TNF-alpha)-induced apoptosis. We found that TNF-alpha induces apoptosis in a time- and dose-dependent manner, whereas IL-4 inhibits TNF-alpha induced and spontaneous apoptosis of biomaterial-adherent macrophages. Blocking experiments and evaluation of shedding of soluble TNF receptor type I (TNF-RI) demonstrated that endogenous TNF-alpha production is responsible for spontaneous apoptosis of biomaterial-adherent cells and that IL-4 inhibits this apoptosis by increasing levels of shedding of soluble TNF-RI. These findings suggest that TNF alpha and IL-4 play key roles in determining the fate of biomaterial-adherent cells and that fusion of macrophages into FBGCs is a mechanism for promoting inflammatory-cell survival on implanted materials. PMID- 11919548 TI - Localization of Reg receptor in rat fundic mucosa. AB - Reg protein has a trophic effect on gastric mucosal cells and pancreatic islets. Recently, the Reg receptor (Reg-R) has been cloned, and Reg-Reg-R interaction has been reported in the pancreas. The aim of this study was to investigate the localization of Reg-R in rat fundic mucosa. Gene expression of Reg-R was investigated with Northern blot analysis, laser capture microdissection coupled with reverse transcription-polymerase chain reaction, and in situ hybridization in the fundic mucosa, and the types of cells expressing this gene were determined. Reg-R mRNA expression was detected mainly in chief cells and parietal cells of the deep layers and faintly in surface epithelial cells and mucous neck cells of the proliferating zone. Our results suggest that regenerating protein may act not only as a regulator of gastric epithelial cell proliferation but also as a modifier of other multiple physiologic functions. PMID- 11919549 TI - Conversion of 7 alpha-hydroxycholesterol to bile acid in human subjects: is there an alternate pathway favoring cholic acid synthesis? AB - Despite the fact that most human subjects synthesize about twice as much cholic acid as chenodeoxycholic acid, available evidence suggests that 7 alpha hydroxycholesterol, the first intermediate in the major pathway for bile acid synthesis, is converted about equally to these two bile acids. Synthesis through the main alternate pathway can not explain this discrepancy because 27 hydroxycholesterol, the first intermediate in that pathway, is converted preferentially to chenodeoxycholic acid. To examine the validity of these contradictory observations, we administered (24-(14)C)-cholic acid and (24-(14)C) chenodeoxycholic acid together with (7 beta-(3)H)-7 alpha-hydroxycholesterol on one occasion and (22,23-(3)H)-27-hydroxycholesterol on a separate occasion to eight normal human subjects. Synthesis of the two primary bile acids was determined by means of standard isotope dilution kinetics of the carbon 14 specific activities of biliary bile acids. Conversion of (7 beta-(3)H)-7 alpha hydroxycholesterol and (22,23-(3)H)-27-hydroxycholesterol to bile acid was calculated from the tritium/carbon 14 ratio in cholic and chenodeoxycholic acid. For synthesis, the mean +/- SEM cholic/chenodeoxycholic ratio was 1.82 +/- 0.26. For apparent conversion of (7 beta-(3)H)-7 alpha-hydroxycholesterol to bile acid, the mean +/- SEM cholic/ chenodeoxycholic ratio was 1.02 +/- 0.09, whereas for (22,23(3)H)-27-hydroxycholesterol, the mean +/- SEM cholic/chenodeoxycholic ratio was 0.38 +/- 0.03. These data imply that, on average, more than 40% of cholic acid in these subjects was synthesized through a pathway that bypassed initial 7 alpha-hydroxylation. However, consideration of all potential candidates for such a pathway raises doubts that any of them contributes substantially to bile acid synthesis. PMID- 11919551 TI - Blood concentrations of lead and erythropoietin. PMID- 11919550 TI - Role of a novel soluble nucleotide phospho-hydrolase from sheep plasma in inhibition of platelet reactivity: hemostasis, thrombosis, and vascular biology. AB - Ecto- and exoenzymes that metabolize extracellular adenosine diphosphate (ADP), the major promoter of platelet activation and recruitment, are of potential clinical importance because they can metabolically prevent excessive thrombus growth. An ecto-ADPase (CD39, NTPDase1) has been identified on endothelial cells. We demonstrate that ADP and adenosine triphosphate (ATP) are rapidly metabolized to adenosine monophosphate (AMP) in sheep plasma at pH 7.4. This hydrolysis is sensitive to P(1), P(5)-di-(adenosine-5') pentaphosphate (Ap(5)A), and ethylene glycol bis (beta-aminoethyl ether) - N, N, N(-), N(-) tetra-acetate (EGTA) but insensitive to tetramisole (an alkaline phosphatase inhibitor). A specific phosphodiesterase substrate, p -nitrophenol-5'-thymidine monophosphate (TMP) (p Nph-5'-TMP), was readily hydrolyzed in sheep plasma at a rate of approximately 0.25 nmol/min/mg protein, and this hydrolysis was inhibited by ADP, ATP, and Ap(5)A. Furthermore, 200-fold purified p -Nph-5'-TMP-hydrolyzing activity also hydrolyzed ATP and ADP directly to AMP. When ADP was preincubated in plasma, its ability to induce platelet aggregation was inhibited in a time-dependent manner. This effect was abolished by Ap(5)A. The inhibitory effects on platelet aggregation correlated with hydrolysis of the ADP in plasma. These data suggest that the endogenous soluble plasma phosphohydrolase metabolizes ATP and ADP by means of cleavage of the alpha-beta-phosphodiester bond of nucleoside 5' phosphate derivatives. This novel biochemical activity inhibits platelet reactivity through hydrolysis of extracellular nucleotides released by activated platelets during (patho)physiological processes, serving a homeostatic and antithrombotic function in vivo. PMID- 11919552 TI - Share and share alike? PMID- 11919553 TI - Unraveling heterochromatin. PMID- 11919554 TI - Untimely ends and new beginnings in mouse cloning. PMID- 11919555 TI - Unifying homology effects. PMID- 11919556 TI - Another cystic mystery solved. PMID- 11919560 TI - The gene mutated in autosomal recessive polycystic kidney disease encodes a large, receptor-like protein. AB - Autosomal recessive polycystic kidney disease (ARPKD) is characterized by dilation of collecting ducts and by biliary dysgenesis and is an important cause of renal- and liver-related morbidity and mortality. Genetic analysis of a rat with recessive polycystic kidney disease revealed an orthologous relationship between the rat locus and the ARPKD region in humans; a candidate gene was identified. A mutation was characterized in the rat and screening the 66 coding exons of the human ortholog (PKHD1) in 14 probands with ARPKD revealed 6 truncating and 12 missense mutations; 8 of the affected individuals were compound heterozygotes. The PKHD1 transcript, approximately 16 kb long, is expressed in adult and fetal kidney, liver and pancreas and is predicted to encode a large novel protein, fibrocystin, with multiple copies of a domain shared with plexins and transcription factors. Fibrocystin may be a receptor protein that acts in collecting-duct and biliary differentiation. PMID- 11919561 TI - Molecular characterization of inter-telomere and intra-telomere mutations in human ALT cells. AB - Telomeres in most immortal cells are maintained by the enzyme telomerase, allowing cells to divide indefinitely. Some telomerase-negative tumors and immortal cell lines maintain long heterogeneous telomeres by the ALT (alternative lengthening of telomeres) mechanism; such tumors are expected to be resistant to anti-telomerase drug therapies. Occasionally telomerase-negative Saccharomyces cerevisiae mutants survive, and 10% of them (type II survivors) have unstable telomeres. As in human ALT+ cells, short telomeres in yeast type II survivors lengthen abruptly; in yeast, this is dependent on the recombination proteins Rad52p and Rad50p. In human cells, ALT involves copying of sequence from a donor to a recipient telomere. We have characterized for the first time a class of complex telomere mutations seen only in ALT+ cells. The mutant telomeres are defined by the replacement of the progenitor telomere at a discrete point (fusion point) with a different telomere repeat array. Among 19 characterized fusion points, one occurred within the first six repeats of the telomere, indicating that these recombination-like events can occur anywhere within the telomere. One mutant telomere may have been involved in a secondary recombination-like mutation event, suggesting that these mutations are sporadic but ongoing in ALT+ cells. We also identified simple intra-allelic mutations at high frequency, which evidently contribute to telomere instability in ALT+ cells. PMID- 11919562 TI - A polymorphic microsatellite that mediates induction of PIG3 by p53. AB - The gene PIG3 is induced by the tumor suppressor p53 but not by p53 mutants unable to induce apoptosis, suggesting its involvement in p53-mediated cell death. Here we show that p53 directly binds and activates the PIG3 promoter, but not through the previously described DNA element. Instead, p53 interacts with a pentanucleotide microsatellite sequence within the PIG3 promoter (TGYCC)n where Y=C or T. Despite its limited similarity to the p53-binding consensus, this sequence is necessary and sufficient for transcriptional activation of the PIG3 promoter by p53 and binds specifically to p53 in vitro and in vivo. In a population of 117 healthy donors from Germany, the microsatellite was found to be polymorphic, the number of pentanucleotide repeats being 10, 15, 16 or 17, and the frequency of alleles 5.1%, 62.0%, 21.4% and 11.5%, respectively. The number of repeats directly correlated with the extent of transcriptional activation by p53. This is the first time that a microsatellite has been shown to mediate the induction of a promoter through direct interaction with a transcription factor. Moreover, this sequence of PIG3 is the first p53-responsive element found to be polymorphic. Inheritance of this microsatellite may affect an individual's susceptibility to cancer. PMID- 11919563 TI - Betting on HSCs. PMID- 11919565 TI - Multilineage development from adult bone marrow cells. PMID- 11919566 TI - Hematopoietic stem cells for transplantation. PMID- 11919567 TI - Therapeutic applications for hematopoietic stem cell gene transfer. PMID- 11919568 TI - Hematopoiesis and stem cells: plasticity versus developmental heterogeneity. AB - Hematopoietic stem cells (HSCs) provide for blood formation throughout the life of the individual. Studies of HSCs form a conceptual framework for the analysis of stem cells of other organ systems. We review here the origin of HSCs during embryological development, the relationship between hematopoiesis and vascular development and the potential plasticity of HSCs and other tissue stem cells. Recent experiments in the mouse have been widely interpreted as evidence for unprecedented transdifferentiation of tissue stem cells. The use of enriched, but impure, cell populations allows for alternative interpretation. In considering these findings, we draw a distinction here between the plasticity of adult stem cells and the heterogeneity of stem cell types that pre-exist within tissues. PMID- 11919569 TI - The aging of lympho-hematopoietic stem cells. AB - The extensive self-renewal capacity of hematopoietic stem cells (HSCs) implies that this cell population may not age and thus may provide undiminished replenishment of blood cells throughout the lifespan of an organism. In contrast, accumulating experimental evidence supports the premise that HSCs show signs of aging and may have a limited functional lifespan. We summarize here the evidence for HSC aging, discuss the possible molecular mechanisms that may be involved and show evidence of a genetic connection between the effects of age on blood-forming cells and the longevity of mice. We speculate that age-related functional decline in adult tissue HSCs limits longevity in mammals. PMID- 11919570 TI - Cellular environment and freedom of gene expression. PMID- 11919571 TI - Lymphoid organs contain diverse cells expressing self-molecules. PMID- 11919573 TI - Killed by leukocytes that don't know when to leave. PMID- 11919574 TI - Type 1 diabetes: virus infection or autoimmune disease? PMID- 11919575 TI - Microbial killing: hold the bleach and pass the salt! PMID- 11919576 TI - Revival of the autoantibody model in rheumatoid arthritis. PMID- 11919577 TI - Class II peptide multimers: promise for type 1A diabetes? PMID- 11919582 TI - Easing the journey back home. PMID- 11919579 TI - Target cell defense prevents the development of diabetes after viral infection. AB - The mechanisms that regulate susceptibility to virus-induced autoimmunity remain undefined. We establish here a fundamental link between the responsiveness of target pancreatic beta cells to interferons (IFNs) and prevention of coxsackievirus B4 (CVB4)-induced diabetes. We found that an intact beta cell response to IFNs was critical in preventing disease in infected hosts. The antiviral defense, raised by beta cells in response to IFNs, resulted in a reduced permissiveness to infection and subsequent natural killer (NK) cell dependent death. These results show that beta cell defenses are critical for beta cell survival during CVB4 infection and suggest an important role for IFNs in preserving NK cell tolerance to beta cells during viral infection. Thus, alterations in target cell defenses can critically influence susceptibility to disease. PMID- 11919583 TI - Rights, wrongs and ignorance. PMID- 11919584 TI - The case for the JASONs. PMID- 11919585 TI - Covert science counsel cut off as nomination row piques Pentagon. PMID- 11919587 TI - Biologists question adult stem-cell versatility. PMID- 11919588 TI - Alternative therapies leave US commission divided. PMID- 11919589 TI - Anger at halal law imperils German research. PMID- 11919590 TI - Hints of bone bounties rile fossil hunters. PMID- 11919591 TI - Babies' cancer screens 'not needed'. PMID- 11919592 TI - Software firm falls victim to shifting bioinformatics needs. PMID- 11919593 TI - Baiting plan to remove fox threat to Tasmanian wildlife. PMID- 11919595 TI - Conflicts of interest: can you believe what you read? PMID- 11919596 TI - Neuroimaging: the sight of two brains talking. PMID- 11919597 TI - The rewards of science extend far beyond publication. PMID- 11919598 TI - The perils of putting career before all else. PMID- 11919599 TI - A quizzical view pays clear dividends. PMID- 11919600 TI - Arab science is not stifled by censorship. PMID- 11919601 TI - Film industry shows how to give credit where due. PMID- 11919602 TI - Science policy: policy, politics and perspective. PMID- 11919607 TI - Conceptual biology: unearthing the gems. PMID- 11919609 TI - Physical chemistry: sculpting ice out of water. PMID- 11919608 TI - Animal behaviour: evolution of suicidal signals. PMID- 11919611 TI - Neurobiology: serotonin sustains serenity. PMID- 11919613 TI - Astrobiology: seeds of life? PMID- 11919614 TI - Cell biology: the ubiquitin connection. PMID- 11919616 TI - Obituary: Sir Raymond Firth (1901-2002). PMID- 11919617 TI - Classical dynamics: spinning eggs--a paradox resolved. AB - If a hard-boiled egg is spun sufficiently rapidly on a table with its axis of symmetry horizontal, this axis will rise from the horizontal to the vertical. (A raw egg, by contrast, when similarly spun, will not rise.) Conversely, if an oblate spheroid is spun sufficiently rapidly with its axis of symmetry vertical, it will rise and spin about the vertical on its rounded edge with its axis of symmetry now rotating in a horizontal plane. In both cases, the centre of gravity rises; here we provide an explanation for this paradoxical behaviour, through derivation of a first-order differential equation for the inclination of the axis of symmetry. PMID- 11919618 TI - Evolutionary biology: lamprey Hox genes and the origin of jaws. AB - The development of jaws was a critical event in vertebrate evolution because it ushered in a transition to a predatory lifestyle, but how this innovation came about has been a mystery. In the embryos of jawed vertebrates (gnathostomes), the jaw cartilage develops from the mandibular arch, where none of the Hox genes is expressed; if these are expressed ectopically, however, jaw development is inhibited. Here I show that in the lamprey, a primitively jawless (agnathan) fish that is a sister group to the gnathostomes, a Hox gene is expressed in the mandibular arch of developing embryos. This finding, together with outgroup comparisons, suggests that loss of Hox expression from the mandibular arch of gnathostomes may have facilitated the evolution of jaws. PMID- 11919619 TI - Biomechanics (Communication arising): prey attack by a large theropod dinosaur. AB - Prey-capture strategies in carnivorous dinosaurs have been inferred from the biomechanical features of their tooth structure, the estimated bite force produced, and their diet. Rayfield et al. have used finite-element analysis (FEA) to investigate such structure-function relationships in Allosaurus fragilis, and have found that the skull was designed to bear more stress than could be generated by simple biting. They conclude that this large theropod dinosaur delivered a chop-and-slash 'hatchet' blow to its prey, which it approached with its mouth wide open before driving its upper tooth row downwards. We argue that this mode of predation is unlikely, and that the FEA results, which relate to an 'overengineered' skull, are better explained by the biomechanical demands of prey capture. Understanding the mechanics of predation is important to our knowledge of the feeding habits of carnivorous dinosaurs and for accurate reconstruction their lifestyles. PMID- 11919621 TI - Ecological responses to recent climate change. AB - There is now ample evidence of the ecological impacts of recent climate change, from polar terrestrial to tropical marine environments. The responses of both flora and fauna span an array of ecosystems and organizational hierarchies, from the species to the community levels. Despite continued uncertainty as to community and ecosystem trajectories under global change, our review exposes a coherent pattern of ecological change across systems. Although we are only at an early stage in the projected trends of global warming, ecological responses to recent climate change are already clearly visible. PMID- 11919622 TI - Serotonin1A receptor acts during development to establish normal anxiety-like behaviour in the adult. AB - Serotonin is implicated in mood regulation, and drugs acting via the serotonergic system are effective in treating anxiety and depression. Specifically, agonists of the serotonin1A receptor have anxiolytic properties, and knockout mice lacking this receptor show increased anxiety-like behaviour. Here we use a tissue specific, conditional rescue strategy to show that expression of the serotonin1A receptor primarily in the hippocampus and cortex, but not in the raphe nuclei, is sufficient to rescue the behavioural phenotype of the knockout mice. Furthermore, using the conditional nature of these transgenic mice, we suggest that receptor expression during the early postnatal period, but not in the adult, is necessary for this behavioural rescue. These findings show that postnatal developmental processes help to establish adult anxiety-like behaviour. In addition, the normal role of the serotonin1A receptor during development may be different from its function when this receptor is activated by therapeutic intervention in adulthood. PMID- 11919623 TI - Racemic amino acids from the ultraviolet photolysis of interstellar ice analogues. AB - The delivery of extraterrestrial organic molecules to Earth by meteorites may have been important for the origin and early evolution of life. Indigenous amino acids have been found in meteorites-over 70 in the Murchison meteorite alone. Although it has been generally accepted that the meteoritic amino acids formed in liquid water on a parent body, the water in the Murchison meteorite is depleted in deuterium relative to the indigenous organic acids. Moreover, the meteoritical evidence for an excess of laevo-rotatory amino acids is hard to understand in the context of liquid-water reactions on meteorite parent bodies. Here we report a laboratory demonstration that glycine, alanine and serine naturally form from ultraviolet photolysis of the analogues of icy interstellar grains. Such amino acids would naturally have a deuterium excess similar to that seen in interstellar molecular clouds, and the formation process could also result in enantiomeric excesses if the incident radiation is circularly polarized. These results suggest that at least some meteoritic amino acids are the result of interstellar photochemistry, rather than formation in liquid water on an early Solar System body. PMID- 11919624 TI - Amino acids from ultraviolet irradiation of interstellar ice analogues. AB - Amino acids are the essential molecular components of living organisms on Earth, but the proposed mechanisms for their spontaneous generation have been unable to account for their presence in Earth's early history. The delivery of extraterrestrial organic compounds has been proposed as an alternative to generation on Earth, and some amino acids have been found in several meteorites. Here we report the detection of amino acids in the room-temperature residue of an interstellar ice analogue that was ultraviolet-irradiated in a high vacuum at 12 K. We identified 16 amino acids; the chiral ones showed enantiomeric separation. Some of the identified amino acids are also found in meteorites. Our results demonstrate that the spontaneous generation of amino acids in the interstellar medium is possible, supporting the suggestion that prebiotic molecules could have been delivered to the early Earth by cometary dust, meteorites or interplanetary dust particles. PMID- 11919625 TI - Exchange-biased quantum tunnelling in a supramolecular dimer of single-molecule magnets. AB - Various present and future specialized applications of magnets require monodisperse, small magnetic particles, and the discovery of molecules that can function as nanoscale magnets was an important development in this regard. These molecules act as single-domain magnetic particles that, below their blocking temperature, exhibit magnetization hysteresis, a classical property of macroscopic magnets. Such 'single-molecule magnets' (SMMs) straddle the interface between classical and quantum mechanical behaviour because they also display quantum tunnelling of magnetization and quantum phase interference. Quantum tunnelling of magnetization can be advantageous for some potential applications of SMMs, for example, in providing the quantum superposition of states required for quantum computing. However, it is a disadvantage in other applications, such as information storage, where it would lead to information loss. Thus it is important to both understand and control the quantum properties of SMMs. Here we report a supramolecular SMM dimer in which antiferromagnetic coupling between the two components results in quantum behaviour different from that of the individual SMMs. Our experimental observations and theoretical analysis suggest a means of tuning the quantum tunnelling of magnetization in SMMs. This system may also prove useful for studying quantum tunnelling of relevance to mesoscopic antiferromagnets. PMID- 11919626 TI - Molecular dynamics simulation of the ice nucleation and growth process leading to water freezing. AB - Upon cooling, water freezes to ice. This familiar phase transition occurs widely in nature, yet unlike the freezing of simple liquids, it has never been successfully simulated on a computer. The difficulty lies with the fact that hydrogen bonding between individual water molecules yields a disordered three dimensional hydrogen-bond network whose rugged and complex global potential energy surface permits a large number of possible network configurations. As a result, it is very challenging to reproduce the freezing of 'real' water into a solid with a unique crystalline structure. For systems with a limited number of possible disordered hydrogen-bond network structures, such as confined water, it is relatively easy to locate a pathway from a liquid state to a crystalline structure. For pure and spatially unconfined water, however, molecular dynamics simulations of freezing are severely hampered by the large number of possible network configurations that exist. Here we present a molecular dynamics trajectory that captures the molecular processes involved in the freezing of pure water. We find that ice nucleation occurs once a sufficient number of relatively long-lived hydrogen bonds develop spontaneously at the same location to form a fairly compact initial nucleus. The initial nucleus then slowly changes shape and size until it reaches a stage that allows rapid expansion, resulting in crystallization of the entire system. PMID- 11919627 TI - Active fluidization of polymer networks through molecular motors. AB - Entangled polymer solutions and melts exhibit elastic, solid-like resistance to quick deformations and a viscous, fluid-like response to slow deformations. This viscoelastic behaviour reflects the dynamics of individual polymer chains driven by brownian motion: since individual chains can only move in a snake-like fashion through the mesh of surrounding polymer molecules, their diffusive transport, described by reptation, is so slow that the relaxation of suddenly imposed stress is delayed. Entangled polymer solutions and melts therefore elastically resist deforming motions that occur faster than the stress relaxation time. Here we show that the protein myosin II permits active control over the viscoelastic behaviour of actin filament solutions. We find that when each actin filament in a polymerized actin solution interacts with at least one myosin minifilament, the stress relaxation time of the polymer solution is significantly shortened. We attribute this effect to myosin's action as a 'molecular motor', which allows it to interact with randomly oriented actin filaments and push them through the solution, thus enhancing longitudinal filament motion. By superseding reptation with sliding motion, the molecular motors thus overcome a fundamental principle of complex fluids: that only depolymerization makes an entangled, isotropic polymer solution fluid for quick deformations. PMID- 11919628 TI - Mantle wedge control on back-arc crustal accretion. AB - At mid-ocean ridges, plate separation leads to upward advection and pressure release partial melting of fertile mantle material; the melt is then extracted to the spreading centre and the residual depleted mantle flows horizontally away. In back-arc basins, the subducting slab is an important control on the pattern of mantle advection and melt extraction, as well as on compositional and fluid gradients. Modelling studies predict significant mantle wedge effects on back-arc spreading processes. Here we show that various spreading centres in the Lau back arc basin exhibit enhanced, diminished or normal magma supply, which correlates with distance from the arc volcanic front but not with spreading rate. To explain this correlation we propose that depleted upper-mantle material, generated by melt extraction in the mantle wedge, is overturned and re-introduced beneath the back-arc basin by subduction-induced corner flow. The spreading centres experience enhanced melt delivery near the volcanic front, diminished melting within the overturned depleted mantle farther from the corner and normal melting conditions in undepleted mantle farther away. Our model explains fundamental differences in crustal accretion variables between back-arc and mid-ocean settings. PMID- 11919629 TI - Determinants of extinction in the fossil record. AB - The causes of mass extinctions and the nature of biological selectivity at extinction events are central questions in palaeobiology. It has long been recognized, however, that the amount of sedimentary rock available for sampling may bias perceptions of biodiversity and estimates of taxonomic rates of evolution. This problem has been particularly noted with respect to the principal mass extinctions. Here we use a new compilation of the amount of exposed marine sedimentary rock to predict how the observed fossil record of extinction would appear if the time series of true extinction rates were in fact smooth. Many features of the highly variable record of apparent extinction rates within marine animals can be predicted on the basis of temporal variation in the amount of exposed rock. Although this result is consistent with the possibility that a common geological cause determines both true extinction rates and the amount of exposed rock, it also supports the hypothesis that much of the observed short term volatility in extinction rates is an artefact of variability in the stratigraphic record. PMID- 11919631 TI - Spatial scale dictates the productivity-biodiversity relationship. AB - The diversity of life is heterogeneously distributed across the Earth. A primary cause for this pattern is the heterogeneity in the amount of energy, or primary productivity (the rate of carbon fixed through photosynthesis), available to the biota in a given location. But the shape of the relationship between productivity and species diversity is highly variable. In many cases, the relationship is 'hump-shaped', where diversity peaks at intermediate productivity. In other cases, diversity increases linearly with productivity. A possible reason for this discrepancy is that data are often collected at different spatial scales. If the mechanisms that determine species diversity vary with spatial scale, then so would the shape of the productivity-diversity relationship. Here, we present evidence for scale-dependent productivity-diversity patterns in ponds. When the data were viewed at a local scale (among ponds), the relationship was hump shaped, whereas when the same data were viewed at a regional scale (among watersheds), the relationship was positively linear. This dependence on scale results because dissimilarity in local species composition within regions increased with productivity. PMID- 11919630 TI - Patterns of colonization in a metapopulation of grey seals. AB - The colonization of a new habitat is a fundamental process in metapopulation biology, but it is difficult to study. The emigration of colonists from established populations might be induced by resource competition owing to high local population density. Migration distances are also important because they determine the frequency and scale of recolonization and hence the spatial scale of the metapopulation. Traditionally, these factors have been investigated with demographic approaches that are labour-intensive and are only possible in amenable species. In many cases, genetic differentiation is minimal, preventing traditional genetic approaches from identifying the source of colonists unambiguously. Here we present a bayesian approach that integrates genetic, demographic and geographic distance data. We apply the method to study the British metapopulation of grey seals, which has been growing at 6% per year over the last few decades. Our method reveals differential recruitment to three newly founded colonies and implicates density-dependent dispersal in metapopulation dynamics by using genetic data. PMID- 11919632 TI - Correlated binocular activity guides recovery from monocular deprivation. AB - Monocular deprivation (MD) has much more rapid and severe effects on the ocular dominance of neurons in the primary visual cortex (V1) than does binocular deprivation. This finding underlies the widely held hypothesis that the developmental plasticity of ocular dominance reflects competitive interactions for synaptic space between inputs from the two eyes. According to this view, the relative levels of evoked activity in afferents representing the two eyes determine functional changes in response to altered visual experience. However, if the deprived eye of a monocularly deprived kitten is simply reopened, there is substantial physiological and behavioural recovery, leading to the suggestion that absolute activity levels, or some other non-competitive mechanisms, determine the degree of recovery from MD. Here we provide evidence that correlated binocular input is essential for such recovery. Recovery is far less complete if the two eyes are misaligned after a period of MD. This is a powerful demonstration of the importance of cooperative, associative mechanisms in the developing visual cortex. PMID- 11919633 TI - Spike-timing-dependent synaptic modification induced by natural spike trains. AB - The strength of the connection between two neurons can be modified by activity, in a way that depends on the timing of neuronal firing on either side of the synapse. This spike-timing-dependent plasticity (STDP) has been studied by systematically varying the intervals between pre- and postsynaptic spikes. Here we studied how STDP operates in the context of more natural spike trains. We found that in visual cortical slices the contribution of each pre-/postsynaptic spike pair to synaptic modification depends not only on the interval between the pair, but also on the timing of preceding spikes. The efficacy of each spike in synaptic modification was suppressed by the preceding spike in the same neuron, occurring within several tens of milliseconds. The direction and magnitude of synaptic modifications induced by spike patterns recorded in vivo in response to natural visual stimuli were well predicted by incorporating the suppressive inter spike interaction within each neuron. Thus, activity-induced synaptic modification depends not only on the relative spike timing between the neurons, but also on the spiking pattern within each neuron. For natural spike trains, the timing of the first spike in each burst is dominant in synaptic modification. PMID- 11919634 TI - Rac function and regulation during Drosophila development. AB - Rac GTPases regulate the actin cytoskeleton to control changes in cell shape. To date, the analysis of Rac function during development has relied heavily on the use of dominant mutant isoforms. Here, we use loss-of-function mutations to show that the three Drosophila Rac genes, Rac1, Rac2 and Mtl, have overlapping functions in the control of epithelial morphogenesis, myoblast fusion, and axon growth and guidance. They are not required for the establishment of planar cell polarity, as had been suggested on the basis of studies using dominant mutant isoforms. The guanine nucleotide exchange factor, Trio, is essential for Rac function in axon growth and guidance, but not for epithelial morphogenesis or myoblast fusion. Different Rac activators thus act in different developmental processes. The specific cellular response to Rac activation may be determined more by the upstream activator than the specific Rac protein involved. PMID- 11919635 TI - Rac GTPases control axon growth, guidance and branching. AB - Growth, guidance and branching of axons are all essential processes for the precise wiring of the nervous system. Rho family GTPases transduce extracellular signals to regulate the actin cytoskeleton. In particular, Rac has been implicated in axon growth and guidance. Here we analyse the loss-of-function phenotypes of three Rac GTPases in Drosophila mushroom body neurons. We show that progressive loss of combined Rac1, Rac2 and Mtl activity leads first to defects in axon branching, then guidance, and finally growth. Expression of a Rac1 effector domain mutant that does not bind Pak rescues growth, partially rescues guidance, but does not rescue branching defects of Rac mutant neurons. Mosaic analysis reveals both cell autonomous and non-autonomous functions for Rac GTPases, the latter manifesting itself as a strong community effect in axon guidance and branching. These results demonstrate the central role of Rac GTPases in multiple aspects of axon development in vivo, and suggest that axon growth, guidance and branching could be controlled by differential activation of Rac signalling pathways. PMID- 11919636 TI - Calmodulin interacts with MLO protein to regulate defence against mildew in barley. AB - In plants, defence against specific isolates of a pathogen can be triggered by the presence of a corresponding race-specific resistance gene, whereas resistance of a more broad-spectrum nature can result from recessive, presumably loss-of regulatory-function, mutations. An example of the latter are mlo mutations in barley, which have been successful in agriculture for the control of powdery mildew fungus (Blumeria graminis f. sp. hordei; Bgh). MLO protein resides in the plasma membrane, has seven transmembrane domains, and is the prototype of a sequence-diversified family unique to plants, reminiscent of the seven transmembrane receptors in fungi and animals. In animals, these are known as G protein-coupled receptors and exist in three main families, lacking sequence similarity, that are thought to be an example of molecular convergence. MLO seems to function independently of heterotrimeric G proteins. We have identified a domain in MLO that mediates a Ca2+-dependent interaction with calmodulin in vitro. Loss of calmodulin binding halves the ability of MLO to negatively regulate defence against powdery mildew in vivo. We propose a sensor role for MLO in the modulation of defence reactions. PMID- 11919638 TI - Crystal structure of DegP (HtrA) reveals a new protease-chaperone machine. AB - Molecular chaperones and proteases monitor the folded state of other proteins. In addition to recognizing non-native conformations, these quality control factors distinguish substrates that can be refolded from those that need to be degraded. To investigate the molecular basis of this process, we have solved the crystal structure of DegP (also known as HtrA), a widely conserved heat shock protein that combines refolding and proteolytic activities. The DegP hexamer is formed by staggered association of trimeric rings. The proteolytic sites are located in a central cavity that is only accessible laterally. The mobile side-walls are constructed by twelve PDZ domains, which mediate the opening and closing of the particle and probably the initial binding of substrate. The inner cavity is lined by several hydrophobic patches that may act as docking sites for unfolded polypeptides. In the chaperone conformation, the protease domain of DegP exists in an inactive state, in which substrate binding in addition to catalysis is abolished. PMID- 11919640 TI - Will public health survive QALYs? PMID- 11919637 TI - A single motif responsible for ubiquitin recognition and monoubiquitination in endocytic proteins. AB - Ubiquitination is a post-translation modification in which ubiquitin chains or single ubiquitin molecules are appended to target proteins, giving rise to poly- or monoubiquitination, respectively. Polyubiquitination targets proteins for destruction by the proteasome. The role of monoubiquitination is less understood, although a function in membrane trafficking is emerging, at least in yeast. Here we report that a short amino-acid stretch at the carboxy-termini of the monoubiquitinated endocytic proteins Eps15 and eps15R is indispensable for their monoubiquitination. A similar sequence, also required for this modification, is found in other cytosolic endocytic proteins, such as epsins and Hrs. These sequences comprise a protein motif, UIM (ref. 6), which has been proposed to bind to ubiquitin. We confirm this for the UIMs of eps15, eps15R, epsins and Hrs. Thus, the same motif in several endocytic proteins is responsible for ubiquitin recognition and monoubiquitination. Our results predict the existence of a UIM:ubiquitin-based intracellular network. Eps15/eps15R, epsins and Hrs may function as adaptors between ubiquitinated membrane cargo and either the clathrin coat or other endocytic scaffolds. In addition, through their own ubiquitination, they may further contribute to the amplification of this network in the endocytic pathway. PMID- 11919641 TI - A novel clinical pharmacy experience for third-year medical students. AB - A novel, clinical curriculum was developed to teach third-year medical students the principles of prescribing for elderly people. The experience involved a didactic session with a community pharmacist and a home visit to assess a senior citizen volunteer who was over age 75 years and was prescribed more than five medications. The medical students completed pre- and postexperience questionnaires to assess knowledge and opinions. Statistical analysis used paired t tests to compare pre- and postknowledge. The percentage agreeing or disagreeing were calculated for Likert opinion responses by using mean summary scores. Pre- and postexperience results were compared using paired t tests. Students showed improved knowledge scores on recognizing drug-drug (P=0.029) and drug-disease interactions (P=0.012). Knowledge on true/false prescribing questions was improved (P=0.005). Students felt that their current curriculum gave insufficient time to prescribing issues, and wanted more education about the use of medications and appropriate prescribing. The majority of students felt that they learned new things (81%), the experience was enjoyable (65%), important topics were covered (71.4%) and they would be more likely to confer with a community pharmacist because of the experience (75%). The novel curriculum described appears to be effective and warrants further evaluation. PMID- 11919643 TI - Comparison of the moisturization efficacy of two vaginal moisturizers: Pectin versus polycarbophil technologies. AB - This study was designed to compare the vaginal deposition and moisturization of two vaginal moisturizers, Summer's Eve (SE), based on pectin, and Replens (Rp), based on polycarbophil, in a double-blind crossover study design. Fifty-one female patients were each randomly assigned to one of two treatment groups. After a one-week washout period, the products were used for two weeks. After another one-week washout period, product assignments were switched. Colposcopy examinations were performed at the beginning and at the end of each product use. Of the forty-seven patients completing the study, 41 (87%) were found to have no vaginal residue after SE vaginal moisturizer, while only 25 (53%) were found to have no vaginal residue after using Rp vaginal moisturizer. No difference in relief of vaginal dryness or in product acceptance was found between the two products. This study shows that the use of SE vaginal moisturizer, based on pectin, resulted in significantly less vaginal residue compared to Rp vaginal moisturizer, based on polycarbophil, and in comparable relief of vaginal dryness. These results strongly suggest that bioadhesion is not important in vaginal moisturizers. PMID- 11919644 TI - Influence of internal structures of hair fiber on hair appearance. I. Light scattering from the porous structure of the medulla of human hair. AB - In this study the influence of the medulla structure on hair appearance was examined. Hair with a porous medulla gave a whitish and lusterless appearance because of light scattering from the fiber center, whereas in the cases without pores, a clear and brilliant appearance was observed. The optical influences of the medulla pores were measured by a spectral goniophotometer, and obtained data were analyzed in terms of the CIE L*a*b* color system. Both contrasts in lightness and apparent color (chroma and hue) decreased in the hair with medulla pores, and the decreases in contrast caused a whitish and lusterless appearance. The distribution of the amount of medulla pores was investigated for Japanese females. The histogram was further analyzed by hair care behavior of individual panelists, and it was found that the pores in the medulla can be generated in a heat-drying process. PMID- 11919645 TI - Optimization and validation of an analytical procedure for the determination of oxidative hair dyes in commercial cosmetic formulations. AB - A method has been developed and validated for the analysis of commonly used intermediates of oxidative hair dyes in commercial cosmetic formulations, including both liquid and cream forms, in dark and blonde shades. The commercial formulations are submitted to extraction by an organic solvent, and the resulting aqueous phase is analyzed by reverse-phase HPLC with a gradient elution and detection with DAD and/or ESI-MS-MS. A spectra library containing 200-400 nm spectra of the target substances and their HPLC retention times has been recorded for the identification. The quantification of the target substances is also performed after spiking of the commercial formulations, using an external calibration. The recoveries obtained are very good for all selected intermediates. The whole procedure has been found to be highly suitable for the identification and quantification of dye intermediates. Also implemented has been a database containing (a) the retention times, (b) the spectral, MS, and MS/MS characteristics of the intermediates, (c) acidity constant values of some intermediates of interest experimentally determined and compared to the available NIST values, (d) the chromatographic conditions used, (e) the behavior towards extraction of dye intermediates, and (f) matrix compounds. PMID- 11919646 TI - Simultaneous determination of alpha and beta hydroxy acids in personal care products by capillary gas chromatography. AB - A simple and rapid procedure is described for the isolation, silylation, and simultaneous capillary gas chromatographic quantitation of alpha hydroxy acids and beta hydroxy acids in various personal care products. The sample is dissolved in acidified N,N-dimethylformamide to simultaneously acidify/extract the hydroxy acids; a portion is then trimethylsilyl derivatized with BSTFA and quantified by capillary gas chromatography (GC) using flame ionization detection. PMID- 11919648 TI - Delayed cardioprotection induced by nitroglycerin is mediated by alpha-calcitonin gene-related peptide. AB - Previous investigations have demonstrated that early preconditioning induced by nitroglycerin is mediated by calcitonin gene-related peptide (CGRP). In the present study, we addressed the question of whether delayed preconditioning induced by nitroglycerin in the rat is related to stimulation of the release and synthesis of CGRP. Sprague-Dawley rats were pretreated with nitroglycerin 24 h before the experiment. The left main coronary artery was occluded for 60 min, followed by 3 h reperfusion. Infarct size, serum creatine kinase activity, serum levels of NO and CGRP and the expression of alpha- and beta-CGRP isoform mRNA in lumbar dorsal root ganglia were measured. Pretreatment with nitroglycerin (60 or 120 microg/kg i.v.) reduced both the infarct size and the release of creatine kinase during reperfusion and caused a significant increase in the expression of alpha-CGRP mRNA, but not beta-CGRP mRNA, concomitantly with an increase in concentrations of NO and CGRP. The increase in CGRP expression preceded the increase in CGRP release. The effects of nitroglycerin were abolished completely by pretreatment with methylene blue (30 mg/kg i.p.), an inhibitor of guanylate cyclase, or capsaicin (50 mg/kg s.c.), which selectively depletes transmitters in capsaicin-sensitive sensory nerves. The present results suggest that the delayed cardioprotection induced by nitroglycerin is mediated mainly by the alpha-CGRP isoform via the NO-cGMP pathway. PMID- 11919649 TI - Diazepam potentiates the effects of endogenous catecholamines on contractility and cyclic AMP levels in rat ventricular myocardium. AB - Diazepam has phosphodiesterase (PDE) inhibitory activity and potentiates the effect of some 3',5'-cyclic adenosine monophosphate (cAMP)-dependent positive inotropic agents. The present study was undertaken to determine whether diazepam enhances the contractile responses and cAMP levels induced by endogenous catecholamines in electrically driven rat right ventricular strips, and the effects are compared with that of the PDE inhibitor 3-isobutylmethylxantine (IBMX). Noradrenaline (10 nM(-1) microM), adrenaline (50 nM-500 microM) and tyramine (5-100 microM) produced concentration-dependent positive inotropic effects that were potentiated by the presence of 10 microM diazepam or IBMX. The diazepam-induced potentiation of the contractile effect of the sympathomimetic agents was not mimicked by 100 microM GABA nor was it antagonized by a 5 microM concentration of the blockers of central and peripheral type benzodiazepine receptors, flumazenil and PK 11195. The beta(2)-adrenergic receptor agonist salbutamol (0.1-300 microM) also produced a concentration-dependent positive inotropic effect which was potentiated by the presence of 10 microM diazepam or 10 microM IBMX. However, the contractile effect of salbutamol, either alone or in the presence of diazepam or IBMX, was not affected by 50 nM ICI 118551, an antagonist of beta(2)-adrenergic receptors, but was virtually abolished by a 0.3 microM concentration of CGP 20712A, an antagonist of beta(1)-adrenergic receptors. Diazepam and IBMX also potentiated the increase in cAMP levels caused by these three sympathomimetic agents in this tissue. [(3)H]Noradrenaline release elicited by electrical stimulation or by tyramine was not affected by diazepam. The results demonstrate that diazepam, like the phosphodiesterase inhibitor IBMX, produces an inotropic and biochemical potentiation of the effects of endogenous catecholamines in rat myocardium. This effect is not due to the release of noradrenaline at the presynaptic level nor is it mediated by beta(2)-adrenergic receptors or benzodiazepine receptors of the central or peripheral type. The effect is probably consequential upon the phosphodiesterase inhibitory activity of diazepam. PMID- 11919650 TI - Antioxidative and myocardial protective effects of L-arginine in oxygen radical induced injury of isolated perfused rat hearts. AB - Oxygen-derived free radicals and oxidants (reactive oxygen intermediates, ROI) have been implicated in cardiovascular diseases. The protective role of nitric oxide (NO) against ROI-mediated tissue injury is not resolved. We tested the effects of exogenous NO, L- and D-arginine and a NO synthase inhibitor on electrolysis-induced cardiac injury and the generation of ROI by electrolysis. Superoxide dismutase (SOD) and catalase were used for comparison. Hearts ( n=7) from male rats (350+/-30 g) were perfused in vitro at 10 ml min(-1) g(-1), ROI generated by electrolysis of the perfusion medium (15 mA, 10 s), and cardiac function and the level of isoluminol-derived chemiluminescence in electrolysed perfusion medium documented for 15 min ( n=4). The ROI-induced maximal reduction of left ventricular developed pressure to 55+/-5% of baseline, and a 2.2+/-0.1 fold rise in coronary perfusion pressure 3 min after electrolysis, were prevented by SOD (50 U ml(-1)), catalase (100 U ml(-1)), S-nitroso- N-acetyl- D,L penicillamine (SNAP, 100 nmol l(-1)); L-arginine (1 mmol l(-1)), N(G)-nitro- L arginine (L-NNA, 200 micromol l(-1)) or D-arginine (1 mmol l(-1)). The effect of L-arginine was concentration dependent. In all cases, the beneficial effects were closely matched by a near-total reduction of ROI in the perfusion medium.We conclude that, besides mimicking or enhancing NO activity, L-arginine and donor derived exogenous NO are cardioprotective by reducing ROI-mediated tissue injury. The protective effect of L-NNA and D-arginine implies that the protection results from a direct chemical interaction between the drug and the oxidizing species. PMID- 11919651 TI - Protective effect of taurine against renal interstitial fibrosis of rats induced by cisplatin. AB - This study was undertaken to investigate the effect of taurine on the infiltration of macrophages and the progression of renal interstitial fibrosis in the kidneys of rats treated with cisplatin (CDDP). Male rats in different groups were treated as follows: (1) saline as control, (2) CDDP and (3) CDDP plus taurine in drinking water. At weeks 2, 4 and 6 after the fifth CDDP injection, we examined platinum content, the kinetics of macrophages and the areas of fibrosis. In the histologic analyses, fibrotic areas were found to have developed around dilated or atrophic tubules in the corticomedullary junction in the kidneys of CDDP-treated rats, while CDDP-plus-taurine-treated rats showed a reduction of the extent and magnitude of damage. These findings reflect the fact that the percentage of fibrotic areas in the kidney of CDDP-plus-taurine-treated rats was significantly lower than in that of CDDP-injected rats throughout the recovery period ( P<0.05). Compared with normal rats, the macrophages in CDDP-injected rats showed significant increases in number at weeks 2, 4 and 6 ( P<0.05). In contrast, the number of macrophages in CDDP-plus-taurine-treated rats was significantly smaller than that of CDDP-injected rats. These results suggested that taurine suppressed the increase of infiltrating macrophages, and led to the attenuation of renal interstitial fibrosis ( P<0.05). PMID- 11919652 TI - Airway relaxant and anti-inflammatory properties of a PDE4 inhibitor with low affinity for the high-affinity rolipram binding site. AB - Inhibitors of phosphodiesterase 4 (PDE4) possess bronchospasmolytic and anti inflammatory properties, which make them very attractive drugs for the treatment of asthma and COPD. Unfortunately, many PDE4 inhibitors also produce central nervous and gastrointestinal side effects, which have limited their clinical application. PDE4 has two binding sites for the archetypal PDE4 inhibitor rolipram, and it has been suggested that binding to the high-affinity rolipram binding site (HARBS) is responsible for the side effects of PDE4 inhibitors. Recently, we have synthesised the PDE4 inhibitor CC3 which shows low affinity to the HARBS. In the present study we investigated the bronchospasmolytic and anti inflammatory properties of this novel compound in comparison to rolipram and the PDE3 inhibitor motapizone. The airway-relaxant properties of the PDE inhibitors were analysed in rat precision-cut lung slices (PCLS) in which airways were contracted by methacholine or in passively sensitised PCLS exposed to ovalbumin. The anti-inflammatory properties were investigated by measuring the release of TNF from endotoxin-treated human monocytes.Up to concentrations of 10 microM none of the PDE inhibitors significantly affected bronchoconstriction elicited by 10 microM methacholine. However, if rolipram or CC3 were given in combination with motapizone, methacholine-induced bronchoconstriction was concentration dependently attenuated. Allergen-induced bronchoconstriction in passively sensitised PCLS was attenuated by CC3 (IC(50) 2.7 microM), rolipram (0.23 microM) and motapizone (8 microM). Combination of equimolar concentrations of motapizone and CC3 (0.34 microM) or rolipram (0.005 microM) showed an additive effect. Endotoxin-induced TNF release from human monocytes was attenuated by all three PDE inhibitors, i.e. CC3 (IC(50) 4.6 microM), rolipram (0.18 microM) and motapizone (5.8 microM). Our findings suggest that PDE4 inhibitors with only low affinity for the HABRS have bronchospasmolytic and anti-inflammatory properties. PMID- 11919653 TI - Concentration-dependent stimulatory and inhibitory effect of troglitazone on insulin-induced fatty acid synthase expression and protein kinase B activity in 3T3-L1 adipocytes. AB - In order to study the effect of the peroxisome proliferator-activated receptor gamma (PPARgamma) agonist troglitazone on the insulin-induced expression of fatty acid synthase (FAS) in adipocytes, we generated a 3T3-L1 cell line stably expressing a FAS reporter gene construct. In this cell line, a low concentration of troglitazone (250 nM) increased the effect of insulin on the FAS promoter activity and the expression of FAS protein about 1.5- to 2-fold. Since the effect of insulin on the expression of FAS is believed to be mediated by activation of protein kinase B (PKB), we investigated the effect of troglitazone on the regulation of PKB. Troglitazone (250 nM) increased the maximal effect of insulin on PKB activity about twofold without significantly affecting its EC(50) (1.4+/ 0.5 nM vs. 2.2+/-0.6 nM in controls). Higher concentrations of troglitazone (> or =1 microM) inhibited both insulin-stimulated PKB activity and expression of FAS. In summary, our data indicate a dual effect of troglitazone on the insulin induced FAS gene expression in 3T3-L1 cells. The therapeutic, stimulatory effect is produced by low concentrations of troglitazone (250 nM), and is presumably mediated by enhanced activation of PKB. PMID- 11919654 TI - Rewarding effects of ethanol and cocaine in mu opioid receptor-deficient mice. AB - To investigate the role of mu opioid receptors in the reinforcing effects of psychotropic drugs, the voluntary ethanol intake and ethanol- and cocaine-induced conditioned place preference in mu opioid receptor-deficient mice and their wild type counterpartners was tested. Moreover, dopamine D1 and D2 receptor binding was measured. It was found that ethanol intake was significantly lower in deficient mice. Conditioned place preference in wild-type animals was induced with 5.0 mg/kg cocaine and this dose was ineffective in the knockouts. In this group conditioned place preference occurred after injection of 10.0 mg/kg cocaine. Cocaine induced a similar increase in locomotor activity in both groups of mice. There was no difference in dopamine D1 receptor binding, whereas dopamine D2 receptor binding was significantly lower in the hippocampus of deficient animals. This suggests that interaction between opioid systems and dopaminergic systems may account for the differences in responding to the drugs. PMID- 11919655 TI - Is Na(+) required for the binding of dopamine, amphetamine, tyramine, and octopamine to the human dopamine transporter? AB - The role of Na(+) in the recognition of blockers by the dopamine transporter is accomodated by a model with a cation site that overlaps with the blocker binding domain, and a distal Na(+) site that interacts with this cation site and perhaps with the blocker binding domain itself. The present study addresses the application of this model to the recognition of substrates by the dopamine transporter, focusing on conditions that should reveal a stimulatory effect, if present, of Na(+) on substrate binding. Recognition was studied via the inhibition of binding of [(3)H]WIN 35,428 (2beta-carbomethoxy-3beta-(4 fluorophenyl) [(3)H]tropane), a cocaine analog, to the human dopamine transporter in human embryonic kidney 293 cells. Little or no changes in binding were noted for dopamine, d-amphetamine, p-tyramine, or dl-octopamine by increasing [Na(+)] from 2 mM to 20 mM with co-varying Br(-), both at pH 7.4 and 7.0. In 74-mM Tris HBr or -HCl, only dopamine and d-amphetamine showed binding increases upon raising Na(+), leveling off with NO(3)(-) or SO(4)(2-) but not Br(-) as anion at approximately 60 mM Na(+), consonant with a partly stimulatory action of Br(-). An Na(+) free, low 5-mM Tris-HEPES buffer was used for studying Na(+) curves truly starting at 0 mM, and, with SO(4)(2-) as the anion, no stimulation of binding by Na(+) was observed. This suggested that the stimulations observed in high (74 mM) Tris(+) buffer by Na(+) were not a direct effect of Na(+) but rather a disinhibitory effect of Na(+) in removing Tris(+) inhibition that depended upon substrate. Tris(+) IC(50) values in Na(+) free buffer were not lower for dopamine and d-amphetamine than p-tyramine and dl-octopamine. No evidence was found for a stronger inhibitory effect of Na(+) for dopamine and dl-octopamine potentially offsetting Tris(+) disinhibition. All results together support the existence of a substrate domain overlapping with a cation site that also binds Tris(+); a distal Na(+) site interacts with this cation site and with the substrate domain by negative allosterism and is additionally impacted by Cl(-). Importantly, interactions between sites vary with the type of substrate, and, in membrane preparations, Na(+) is not required for, or stimulatory to, the binding of any of the four substrates studied unlike the binding of the cocaine analog WIN 35,428. PMID- 11919656 TI - Amino acids involved in differences in the pharmacological profiles of the rat and human noradrenaline transporters. AB - It has been suggested from a previous study in our laboratory that differences in the pharmacology of the species variants of the noradrenaline transporter (NET) are the result of four non-conservative amino acid exchanges from the total of 26 amino acids that are divergent between the rat NET (rNET) and human NET (hNET). The aim of this study was to examine the effects of changing the rNET at each of these four amino acid residues, which markedly alter local charge distribution, to the amino acid found in hNET.Site-directed mutagenesis was used to create mutant cDNAs from rNET cDNA. The mutant NETs (rK7D, rE62K, rK375N and rR612Q), rNET and hNET were expressed in transiently transfected COS-7 cells to determine the effects of the mutations on the differing pharmacological properties of the species variants. The ratios of V(max) for noradrenaline uptake and B(max) for nisoxetine binding (which are a measure of the turnover number of the transporter, i.e. the number of transport cycles per min) were greater for rNET and rR612Q than for hNET, rK7D, rE62K and rK375N. The K(m) of noradrenaline was lower for hNET, rK7D, rE62K and rK375N than for rNET or rR612Q. There were no differences between the K(i) values for inhibition of noradrenaline uptake by nisoxetine for rNET, hNET or the mutants, but the K(i) values of cocaine were lower for hNET, rE62K and rR612Q than rNET or rK375N.Hence, the study showed that: (1) the aspartate 7, lysine 62 and asparagine 375 amino acid residues are important in determining the lower substrate translocation by hNET than rNET; (2) the aspartate 7 and lysine 62 residues in the N-terminus of hNET determine the higher affinities of substrates for the hNET than the rNET; and (3) the lysine 62 and glutamine 612 residues in the N- and C-termini, respectively, of hNET are determinants of the higher cocaine affinity for the hNET than rNET. PMID- 11919657 TI - Time-of-day dependent pharmacodynamic and pharmacokinetic profiles of caffeine in rats. AB - This study aims to investigate the effects of caffeine on the daily rhythms of heart rate (HR), body temperature (BT) and locomotor activity (LA) in rats in relation to time-of-day of administration, as well as their possible mechanisms, particularly related to caffeine pharmacokinetics. During the pharmacodynamic study, HR, BT and LA were measured every 10 min by radiotelemetry and analysed by Cosinor. This study was divided into three periods: a control period P1, a treatment period P2 and a recovery period P3. During P2, rats of the morning group ( M(tel)) received a 25 mg/kg s.c. dose of caffeine at 08.00 while rats of the evening group ( E(tel)) received the same dose of caffeine at 20.00. The pharmacokinetic study was conducted in parallel with the telemetric study and was divided into two periods: a control period P1, and a treatment period P2. During P2, animals of the morning ( M(pk)) and the evening ( E(pk)) groups received the same treatment as the animals of the telemetric study. At the last day of P2, blood samples were drawn 0.25, 0.5, 1, 2, 4, 8, 12 and 24 h after the last morning and the last evening administration in order to determine the pharmacokinetics of M(pk) and E(pk). Our results showed that morning administration of caffeine suppressed the daily rhythmicity of LA and modified the mesors and amplitudes of the HR and BT daily rhythms, while the evening administration did not suppress the daily rhythm of LA, but altered the mesors, amplitudes and acrophases of the three rhythms, indicating a chronopharmacological effect. With respect to the pharmacokinetic effects, the area under the curve (AUC) was significantly lower in rats of E(pk) compared with M(pk), due to an increase of the total plasma clearance and the volume of distribution. Our data suggest that the chronopharmacokinetic effects of caffeine may explain, at least in part, the observed caffeine-induced modifications on the daily rhythms. PMID- 11919658 TI - Analysis of the effect of cannabinoids on GABAergic neurotransmission in the substantia nigra pars reticulata. AB - The CB(1) cannabinoid receptor is widely distributed in the central nervous system. The substantia nigra pars reticulata (SNR) belongs to the brain regions with the highest density of CB(1) receptors. According to anatomical studies, most of the CB(1) receptors in the SNR are localized on terminals of striatonigral GABAergic neurons. The aim of the present study was to clarify the function of these receptors.Electrophysiological properties of SNR neurons were studied in brain slices with the patch-clamp technique. Inhibitory postsynaptic currents (IPSCs) were elicited in parasagittal slices by electrical stimulation in the internal capsule. The mixed CB(1)/CB(2) cannabinoid receptor agonist WIN55212-2 (1 microM and 10 microM) concentration dependently decreased the amplitude of IPSCs. CP55940, another mixed CB(1)/CB(2) cannabinoid receptor agonist, also lowered IPSC amplitude. Superfused alone, the CB(1)-selective antagonist SR141716A (1 microM) increased the amplitude of IPSCs. In interaction experiments, SR141716A (1 microM) prevented the inhibition produced by WIN55212-2 (1 microM). WIN55212-2 (1 microM) had no effect on GABAergic currents elicited by ejection of muscimol (1 mM) to the surface of the slices. WIN55212-2 (10 microM) did not influence the frequency and amplitude of spontaneously occurring IPSCs (sIPSCs) and the firing rate of SNR neurons. The results show that activation of CB(1) cannabinoid receptors inhibits GABAergic neurotransmission in the SNR. The likely mechanism is presynaptic inhibition of GABA release, since cannabinoids had no effects on currents evoked by direct stimulation of GABA(A) receptors by muscimol and on the amplitude of sIPSCs. The enhancement of IPSCs by the cannabinoid antagonist probably reflects continuous inhibition of GABAergic neurotransmission by an endogenous cannabinoid. SNR neurons receive GABAergic input from three sources: from the corpus striatum, the globus pallidus and from neighbouring SNR neurons. The observed inhibition of GABAergic neurotransmission was probably due to depression of the transmission between striatonigral axons and SNR neurons. No direct actions of cannabinoids on SNR neurons were observed in addition to this synaptic effect. PMID- 11919659 TI - Marian W. Fischman: 1939-2001. PMID- 11919660 TI - Concurrent administration of diethyldithiocarbamate and 4-methylpyrazole enhances ethanol-induced locomotor activity: the role of brain ALDH. AB - RATIONALE: It has been proposed that brain aldehyde dehydrogenase (ALDH) plays a role in the modulation of some psychopharmacological effects of ethanol. Diethyldithiocarbamate (DDTC), an ALDH inhibitor, elevates blood acetaldehyde levels in the presence of ethanol. Concurrent administration with 4 methylpyrazole (4-MP), an alcohol dehydrogenase inhibitor, prevents peripheral accumulation of acetaldehyde by DDTC. OBJECTIVE: To investigate the effects of concurrent DDTC and 4-MP administration on ethanol-induced locomotor activity in mice. METHODS: Mice were pretreated IP with saline (S+S) or 4-MP (10 mg/kg) (S+4 MP), then received IP injections of ethanol (0, 0.8, 1.6, 2.4, 3.2 and 4 g/kg) prior to testing in the open field. RESULTS: Pretreatment with 4-MP does not modify the spontaneous or ethanol-induced locomotor activity. In the second experiment, the DDTC (114, 228 and 456 mg/kg) and 4-MP (DDTC+4-MP) were administered 8 h prior to testing locomotor activity in the open field. Animals were then treated with ethanol (0, 0.8, 1.6, 2.4, 3.2 and 4 g/kg), and placed in open field chambers. The locomotor activity of animals pretreated with DDTC and 4 MP was significantly enhanced here compared to groups S+S and S+4-MP. These effects cannot be attributed to elevated blood acetaldehyde levels, as pretreatment with 4-MP prevented peripheral accumulation of acetaldehyde. CONCLUSIONS: These data suggest that brain ALDH may contribute to the effects of ethanol on locomotor activity. This role of the enzyme ALDH in some of the psychopharmacological effects of ethanol may be a result of its ability to regulate levels of acetaldehyde in brain. PMID- 11919661 TI - Effects of buprenorphine sublingual tablet maintenance on opioid drug-seeking behavior by humans. AB - RATIONALE: Buprenorphine can decrease opioid self-administration by humans and animals, but its ability to decrease drug-seeking behavior and craving (i.e. motivational measures) among outpatient volunteers using clinically relevant dosing schedules has not been extensively studied. OBJECTIVES: We investigated whether daily versus alternating-day administration of high versus low buprenorphine doses influenced choice of, and operant responding for, hydromorphone versus money. METHODS: Fourteen heroin-dependent outpatients were maintained under four buprenorphine sublingual tablet (double blind) dose conditions using a within-subject, randomized crossover design. All participants received, for 2 weeks each, buprenorphine doses of 2 mg daily, 4 mg/placebo on alternating days, 16 mg daily, and 32 mg/placebo on alternating days. In each laboratory test session, participants chose between money ($2/choice) and drug (1/8 of total hydromorphone, 4 or 24 mg IM in different sessions) alternatives using an eight-trial non-independent progressive ratio schedule (FR 100, 200,.12,800). The drug dose and money amount earned was delivered after the end of the 2.5-h work period. RESULTS: Hydromorphone 24 mg was more reinforcing than 4 mg. Higher versus lower average buprenorphine doses (regardless of daily versus alternate-day schedule) significantly decreased hydromorphone 24 mg choice and increased money choice. Baseline heroin craving questionnaire scores predicted drug choice, and craving scores were significantly decreased by high-dose buprenorphine. CONCLUSIONS: High-dose buprenorphine attenuated opioid drug seeking behavior, heroin craving self-reports and increased sensitivity to alternative reinforcement. These beneficial effects were retained when high-dose buprenorphine was administered on alternate days. PMID- 11919662 TI - Fluoxetine, but not other selective serotonin uptake inhibitors, increases norepinephrine and dopamine extracellular levels in prefrontal cortex. AB - RATIONALE: The selective serotonin uptake inhibitor (SSRI) fluoxetine has been shown to not only increase the extracellular concentrations of serotonin, but also dopamine and norepinephrine extracellular concentrations in rat prefrontal cortex. The effect of other SSRIs on monoamine concentrations in prefrontal cortex has not been thoroughly studied. OBJECTIVE: The aim of this study was to compare the ability of five systemically administered selective serotonin uptake inhibitors to increase acutely the extracellular concentrations of serotonin, norepinephrine and dopamine in rat prefrontal cortex. METHODS: The extracellular concentrations of monoamines were determined in the prefrontal cortex of conscious rats using the microdialysis technique. RESULTS: Fluoxetine, citalopram, fluvoxamine, paroxetine and sertraline similarly increased the extracellular concentrations of serotonin from 2- to 4-fold above baseline. However, only fluoxetine produced robust and sustained increases in extracellular concentrations of norepinephrine and dopamine after acute systemic administration. Fluoxetine at the same dose blocked ex vivo binding to the serotonin transporter, but not the norepinephrine transporter, suggesting that the increase of catecholamines was not due to non-selective blockade of norepinephrine uptake. Prefrontal cortex extracellular concentrations of fluoxetine at the dose that increased extracellular monoamines were 242 nM, a concentration sufficient to block 5-HT(2C) receptors which is a potential mechanism for the fluoxetine-induced increase in catecholamines. CONCLUSION: Amongst the SSRIs examined, only fluoxetine acutely increases extracellular concentrations of norepinephrine and dopamine as well as serotonin in prefrontal cortex, suggesting that fluoxetine is an atypical SSRI. PMID- 11919663 TI - Effects of chronic administration of the D1 receptor partial agonist SKF 77434 on cocaine self-administration in rhesus monkeys. AB - RATIONALE: Dopamine D1 receptor partial agonists have been proposed as candidate medications for the treatment of cocaine dependence. However, there currently is scant information regarding how chronic exposure to D1 agonists may modify behavioral effects of cocaine and, especially, whether tolerance develops to their effects on cocaine self-administration. OBJECTIVE: The present studies were conducted to evaluate the effects of chronic treatment with the D1 receptor partial agonist SKF 77434 on IV cocaine self-administration in rhesus monkeys. METHODS: A protocol was developed to rapidly evaluate the effects of chronic drug exposure on extinction behavior, threshold dose of self-administered cocaine, and the dose-effect function for cocaine self-administration behavior. Monkeys performed in daily sessions of IV cocaine self-administration under a fixed-ratio schedule of reinforcement and food presentation under either a fixed-ratio or fixed-interval schedule of reinforcement. When both types of performance were stable, chronic exposure to SKF 77434 followed with month-long regimens of IV treatment with each of two or three dosages. RESULTS: The effects of SKF 77434 were dose-related. Exposure to 1.0 mg/kg per day of SKF 77434 yielded a moderate and persistent rightward shift in the descending portion of the dose-effect function for cocaine self-administration but did not alter the threshold dose and did not disrupt either extinction behavior or food-maintained performance. An increase in dosage to 3.2-5.6 mg/kg per day displaced the dose-effect function for cocaine self-administration downward from its prechronic position, altered threshold dose values, and disrupted food-maintained performance. CONCLUSIONS: Chronic treatment with D1 receptor partial agonists produced dose-dependent effects on cocaine self-administration that may be relevant to their further evaluation as candidate medications for the treatment of cocaine dependence. PMID- 11919664 TI - Dopamine antagonists alter response allocation but do not suppress appetite for food in rats: contrast between the effects of SKF 83566, raclopride, and fenfluramine on a concurrent choice task. AB - RATIONALE: Dopamine is important for enabling organisms to overcome work-related response costs. One way of investigating this function has been with concurrent choice procedures using food reinforcement. In the present study, rats were given a choice between pressing a lever for preferred Bioserve pellets, or approaching and consuming a less-preferred laboratory chow that was concurrently available. In previous work with this task, dopamine antagonists and accumbens dopamine depletions decreased lever pressing but increased chow consumption. OBJECTIVE: The present study assessed three drugs (two dopamine antagonists and one appetite suppressant) using the lever pressing/chow feeding task. RESULTS: Under baseline conditions, rats pressed the lever at high rates (1,300-1,500 responses) to obtain the preferred food, and little of the laboratory chow was eaten (1-2 g). Selective D1 and D2 antagonists (SKF 83566 and raclopride) reduced fixed ratio 5 lever pressing, but substantially increased chow consumption. In contrast, the serotonergic appetite suppressant fenfluramine reduced both lever pressing and chow consumption. With the dopamine antagonists, lever pressing and chow consumption were inversely correlated across treatments, while these two measures were unrelated in the fenfluramine experiment. CONCLUSIONS: Dopamine antagonists and accumbens dopamine depletions do not simply reduce appetite. Rats with accumbens dopamine depletions, or rats treated with low doses of selective or non selective dopamine antagonists, remain directed toward the acquisition and consumption of food. These results demonstrate that fundamental aspects of food reinforcement are left intact after treatment with low doses of dopamine antagonists. PMID- 11919665 TI - Modulation of the critical flicker fusion effects of serotonin reuptake inhibitors by concomitant pupillary changes. AB - RATIONALE: Serotonin reuptake inhibitors (SSRIs) have been attributed CNS activating properties based on their ability to elevate the Critical Flicker Fusion (CFF) threshold. However, such an interpretation may be questioned since CFF elevations may also be due to SSRI-induced increases in pupil diameter. OBJECTIVES: The effect of pupillary changes on CFF assessment following SSRI administration was investigated in a double blind, crossover study. METHODS: During three periods of 15 days, 21 healthy men and women (30-50 years) received sertraline (50 mg on days 1-8, 100 mg on days 9-15), citalopram (20 mg on days 1 8, 40 mg on days 9-15) and placebo. Assessments were done on days 1, 8 and 15 and consisted of pupillary measurements and CFF assessments with and without pupillary control (a 2-mm artificial pupil) using the Leeds Psychomotor Tester. RESULTS: Both SSRIs induced an acute and steady increase in pupil diameters. CFF thresholds were depressed following acute administration of sertraline and citalopram, but this was only apparent when a control was made for the pupillary changes. No CFF effects were seen at day 8, but CFF was again reduced at day 15, with and without control for pupil size. CONCLUSIONS: Mydriasis masked the detrimental effects of both SSRIs on CFF during the acute assessments. Our results raise questions regarding the validity of the assessment of the behavioural toxicity of SSRIs based on CFF measurements without ample control for pupil size, especially when these concern acute measurements. PMID- 11919666 TI - Investigation into the significance of task difficulty and divided allocation of resources on the glucose memory facilitation effect. AB - RATIONALE: Memory for a list of 20 words can be enhanced by preceding learning with consumption of 25 g glucose rather than an equally sweet aspartame solution. In previous studies, participants performed a secondary hand-movement task during the list-learning phase. OBJECTIVE: The present placebo-controlled, double-blind study examined whether the additional cognitive load created by a secondary task is a crucial feature of the glucose memory facilitation effect. METHODS: The effect of glucose administration on word recall performance in healthy young participants was examined under conditions where the primary memory task and a secondary task were competing for cognitive resources (across a range of secondary tasks), and where task difficulty was increased but dual task-mediated competition for cognitive resources did not exist. Measures of non-verbal and working memory performance were also compared under the different glycaemic conditions (glucose versus aspartame drinks). RESULTS: In the present study, a beneficial effect of glucose on memory was detected after participants encoded a 20-word list while performing a secondary task, but not when participants encoded the list without a secondary task, nor when the 20 target words were intermixed with 20 non-target words (distinguished by gender of speaker). In addition, glucose significantly enhanced performance on spatial and working memory tasks. CONCLUSION: The data indicate that possible "depletion" of episodic memory capacity and/or glucose-mediated resources in the brain due to performing a concomitant cognitive task might be crucial to the demonstration of a glucose facilitation effect. Possible implications regarding underlying cognitive and physiological mechanisms are discussed in this article. PMID- 11919667 TI - Mapping murine loci for physical dependence on ethanol. AB - RATIONALE: Alcoholism is associated with withdrawal (physical dependence), tolerance, or a maladaptive pattern of alcohol (ethanol) use. The well-documented difference in susceptibility to withdrawal after chronic ethanol exposure between the C57BL/6J and DBA/2J mouse strains provides an excellent starting point for dissecting genetic influences involved in physical dependence on ethanol. A quantitative trait locus (QTL) identifies the genomic location of a gene (or genes) affecting a trait of interest. OBJECTIVES: A genome-wide QTL mapping study was carried out to dissect the multifactorial nature of withdrawal after chronic ethanol exposure using 400 B6D2F2 mice. METHODS: To induce physical dependence, we used a standard paradigm in which mice were exposed to ethanol vapor for 72 h. The mice were then tested hourly for handling-induced convulsions (HICs) for 10 h and at hours 24 and 25. Ethanol withdrawal severity was first computed as the area under the 25-h HIC curve. Separate regression residuals were then calculated that corrected for individual differences in blood ethanol concentration at the time of withdrawal and baseline HIC severity (i.e. before ethanol exposure). RESULTS: Statistical mapping yielded significant evidence ( P<0.00005) for QTLs on chromosomes 19 and distal 1 that account for 45% of the genetic variance in ethanol withdrawal severity. The F2 results also provide supporting evidence for a sex-limited QTL on chromosome 13, and QTLs on chromosomes 4 and proximal 1, which may account for an additional 38% of the genetic variance. The distal chromosome 1 QTL is a locus of major effect, accounting for 26% of the genetic variance. Experiments using two congenic strains more precisely mapped this QTL. CONCLUSIONS: The QTLs map near candidate genes involved in neurosteroid biosynthesis and signal transduction. Syntenic homology between human and mouse chromosomes suggests that genes related to physical dependence on ethanol may localize to human chromosome regions 10q23-q26, 1q21-q43, 2q11-q32, 5p15/5q14 q21, and 9p24-p22. PMID- 11919668 TI - Ondansetron reduces the craving of biologically predisposed alcoholics. AB - RATIONALE: Early onset alcoholics (EOA) differ from late onset alcoholics (LOA) by having greater serotonergic abnormality, familial history, and a range of antisocial behaviors. Previously, we showed that ondansetron, a selective 5-HT3 antagonist, effectively treated EOA. Proximate motivational drives such as craving could have determined drinking behavior. We therefore investigated whether ondansetron treatment would reduce alcohol craving significantly among EOA. OBJECTIVES: We tested the hypothesis that the craving outcomes of EOA, compared with LOA, would be differentially improved by ondansetron. We also tested the prediction that craving would be significantly correlated with drinking behavior. METHODS: We studied a cohort of 253 out of 321 enrolled alcohol dependent subjects. These 253 subjects were entered into a 1-week lead-in single-blind placebo period followed by 11 weeks of double-blind outpatient treatment. Study design was a 2 (EOA versus LOA)x 4 medication dose (placebo, or ondansetron 1, 4, or 16 microg/kg b.i.d)x 13 (visits) factorial analysis of variance. Craving was measured at each visit using seven visual analogue scales. Subjects received 12 weekly sessions of standardized group cognitive behavioral therapy. RESULTS: Data reduction by factor analysis of the visual analog scale items yielded one dimension, overall craving. Ondansetron 4 microg/kg b.i.d. reduced overall craving significantly among EOA. In contrast, ondansetron (1 microg/kg b.i.d.) increased craving significantly in LOA. Decreased overall craving was positively correlated with reduced drinking and negatively associated with increased abstinence. CONCLUSIONS: Compared with placebo, ondansetron (4 microg/kg b.i.d.) was associated with significant reductions in overall craving in EOA but not LOA, presumably by ameliorating serotonergic abnormality. PMID- 11919669 TI - Distal and proximal pre-exposure to ethanol in the place conditioning task: tolerance to aversive effect, sensitization to activating effect, but no change in rewarding effect. AB - RATIONALE: The literature offers many examples of tolerance to ethanol's inhibitory/depressant effects and sensitization to its activating effects. There are also many examples of tolerance to ethanol's aversive effects as measured in the conditioned taste aversion and conditioned place aversion (CPA) procedures. However, there are very few demonstrations of either tolerance or sensitization to ethanol's rewarding or reinforcing effects. OBJECTIVE: The present studies were designed to examine effects of two forms of ethanol pre-exposure (distal or proximal) on ethanol's rewarding and aversive effects as indexed by the place conditioning procedure. METHOD: Male inbred (DBA/2J) mice were exposed to ethanol (2 g/kg IP) in an unbiased place conditioning procedure that normally produces either conditioned place preference (CPP) (ethanol injection before CS exposure) or CPA (ethanol injection after CS exposure). In the distal pre-exposure studies (experiments 1 and 2), mice initially received a series of four ethanol injections (0, 2, or 4 g/kg) in the home cage at 48-h intervals during the week before place conditioning. In the proximal pre-exposure studies (experiments 3 4), mice were injected with ethanol 65 min before (experimental groups) or 65 min after (control groups) each paired ethanol injection on CS+ trials. RESULTS: Distal pre-exposure produced a robust sensitization to ethanol's activating effect, whereas proximal pre-exposure generally reduced the activation normally produced by the paired ethanol injection. Both forms of pre-exposure interfered with CPA, but had no effect on CPP. CONCLUSIONS: These studies suggest that both forms of pre-exposure reduced ethanol's aversive effect, but had no impact on ethanol's rewarding effect. In general, the detrimental effects of pre-exposure on CPA are explained best in terms of a reduction in ethanol's efficacy as an aversive unconditioned stimulus (i.e. tolerance), although explanations based on other types of associative interference are also possible. The failure to affect CPP with pre-exposure treatments that reduced or eliminated CPA suggests that these behaviors are mediated by independent, motivationally opposite effects of ethanol. Moreover, these results indicate dissociation between sensitization to ethanol's locomotor activating effect and changes in its rewarding effect. To the extent that motivational processes measured by CPP and CPA normally contribute to ethanol drinking, the present findings suggest that increases in ethanol intake seen after chronic ethanol exposure are more likely caused by tolerance to ethanol's aversive effect rather than sensitization to its rewarding or reinforcing effect. PMID- 11919670 TI - Basolateral amygdala inactivation abolishes conditioned stimulus- and heroin induced reinstatement of extinguished heroin-seeking behavior in rats. AB - RATIONALE: Drug-paired stimuli elicit drug craving and relapse in addicts and drug-seeking behavior in rats. The functional integrity of the basolateral amygdala (BLA) is necessary for reinstatement of cocaine-seeking behavior elicited by cocaine-conditioned stimuli, but not by cocaine itself. It is unclear, however, whether the BLA plays a similar role in reinstatement of heroin seeking behavior. OBJECTIVES: To this end, we examined the effects of tetrodotoxin (TTX)-induced inactivation of the BLA on conditioned and heroin primed reinstatement of extinguished heroin-seeking behavior. METHODS: Rats were trained to press a lever for IV infusions of heroin (maintenance dose of 25 microg/infusion) paired with presentations of a light-tone stimulus complex during daily 3-h sessions. Responding was then allowed to extinguish prior to reinstatement testing. Reinstatement of extinguished heroin-seeking behavior (i.e. lever pressing in the absence of heroin reinforcement) was measured in the presence of response-contingent presentation of the heroin-paired stimulus complex alone and then following TTX (5 ng/0.5 microl per side) or vehicle infused into the BLA. In a separate group of rats, reinstatement was measured after saline injection (SC) and then following heroin priming (0.25 mg/kg, SC) with TTX or vehicle infused into the BLA. RESULTS: Both response contingent presentation of the stimulus complex and heroin priming significantly reinstated extinguished heroin-seeking behavior, and BLA inactivation abolished the ability of the heroin-paired stimuli and of heroin priming to reinstate responding. CONCLUSIONS: These findings suggests that the BLA is a critical component of the neural circuitry that mediates conditioned and heroin-induced reinstatement of heroin-seeking behavior. Furthermore, different neural substrates may mediate drug-primed relapse to cocaine versus heroin-seeking behavior. PMID- 11919671 TI - Effects of tryptophan depletion on brain potential correlates of episodic memory retrieval. AB - RATIONALE: Neuropsychological impairments in depressive illness may be secondary to proposed serotonergic abnormalities. Acute tryptophan depletion (ATD) in healthy subjects impairs episodic memory, but the mechanism of this is unclear. OBJECTIVES: To examine the effects of ATD on the neural correlates of episodic memory retrieval in healthy subjects. METHODS: Fourteen healthy men were given an amino acid cocktail drink with or without tryptophan, in a double blind, crossover design. Event related potentials (ERPs) were recorded during a well validated episodic memory task performed 5 h after drink ingestion. Subjects listened to words spoken in a male or female voice. At test, old and new words were presented visually; subjects judged whether words were old or new, and if old, the gender of the voice at study. RESULTS: ATD led to an 84+/-5% reduction in plasma free tryptophan concentrations, and significantly impaired episodic memory recall. ERP recordings demonstrated previously reported left parietal and right frontal "old/new" differences for ERPs to items associated with accurate episodic memory retrieval versus correctly rejected new items. ATD increased ERP voltage between 500 and 1400 ms post-stimulus particularly over posterior regions of the scalp, but there was no interaction with item type. Topographical analysis of the old/new difference revealed no significant treatment by site interaction. CONCLUSIONS: ATD impairs episodic memory recall with no effect on the magnitude or topography of the neural correlates of retrieval in healthy subjects. This suggests that the effects of ATD on recall may reflect an impairment of memory encoding and/or consolidation. PMID- 11919674 TI - Regulation of glycolysis by casein kinase I (Rag8p) in Kluyveromyces lactis involves a DNA-binding protein, Sck1p, a homologue of Sgc1p of Saccharomyces cerevisiae. AB - The casein kinase I (Rag8p) of Kluyveromyces lactis has previously been shown to regulate the transcription of the low-affinity glucose transporter gene RAG1. To study this regulation, we have isolated multicopy suppressors of the rag8 mutation. One of them, SCK1 (suppressor of casein kinase), was characterised. The predicted product of the gene has a DNA-binding signature of the basic-helix-loop helix type. It has an overall identity of 38% with Sgc1p (Tye7p) of Saccharomyces cerevisiae. The sck1 null mutant exhibited a Rag- phenotype (which indicates a reduced flux of glycolysis) that can be complemented by the SGC1 gene of S. cerevisiae. The level of transcription of several glycolytic genes, including RAG1, was reduced about twofold in glucose media in the sck1 null mutant. Moreover, in a rag8 mutant, the expression of SCK1 was strongly affected. Altogether, the results suggest that the regulation of glycolysis by casein kinase I involves, at least in part, Sck1p in K. lactis. PMID- 11919675 TI - Dynamics of the mitochondrial genome during Podospora anserina aging. AB - Senescence in Podospora anserina is always correlated with extrachromosomal mitochondrial DNA amplification (senDNA). Here we report a quantitative kinetic analysis of the molecular events that occur in the mitochondrial DNA of several wild-type cultures during aging. For each culture, the amplification of senDNA molecules and the modifications of the mitochondrial chromosome are analyzed at different ages and in relation with two age-related parameters: growth rate and fertility. We find senDNAs exponentially amplified from the germination state and particular regions of the mitochondrial chromosome preferentially lost only in the pre-senescent state, i.e. a few centimeters before the growth arrest of the mycelium. This late loss of information, concomitant with the first phenotypic expression of senescence (loss of fertility), begins in the regions from which the senDNAs originate and culminates with the growth arrest of the mycelium. PMID- 11919676 TI - Cloning and characterization of multiple glycosyl hydrolase genes from Trichoderma virens. AB - Trichoderma virens is a widely distributed soil fungus that is parasitic on other soil fungi. The mycoparasitic activity of T. virens is correlated with the production of numerous antifungal activities, including the secretion of a considerable repertoire of fungal cell wall-degrading enzymes. Here, we report the characterization of a diverse set of chitinase and glucanase genes from T. virens. In each case, full-length genomic clones were isolated and characterized, while sequencing of the corresponding cDNA clones and manual annotation provided a basis for establishing gene structure. Based on homology of the deduced amino acid sequences, we have identified three members of the 42Kd endochitinase gene family, two 33Kd exochitinases, two exochitinases with homology to N acetylglucosaminidases, and three glucanase genes predicted to encode beta-1,3- and beta-1,6-proteins. The majority of these genes appear to encode signal peptides, suggesting an extracellular location for the corresponding proteins. Despite their overall similarity, the 42Kd class of chitinases can be subdivided, based on the presence of distinct N-terminal domains, suggesting that the proteins may have distinct cellular roles, while Northern blot analysis confirms that these genes possess distinct patterns of gene regulation. Similarly, one of the 33Kd chitinase genes is unique, because it is predicted to encode a protein C terminus with high homology to the conserved family I cellulose-binding domain. The expression patterns of the chitinase genes were analyzed in both a wild-type strain and a strain disrupted for the major 42Kd chitinase gene of T. virens. The results of these transcript analyses, together with enzymatic assay of the extracellular proteins, suggest interdependent regulation of this important gene family in T. virens. PMID- 11919677 TI - The pipg1 gene of the oomycete Phytophthora infestans encodes a fungal-like endopolygalacturonase. AB - Endopolygalacturonases (endoPGs) are plant cell wall-degrading enzymes that have been implicated in the invasion of plant tissue by pathogenic microbes. EndoPGs have been described from bacteria, plants, insects and numerous species of phytopathogenic fungi. In this study, we describe the first endoPG sequence from oomycetes, a unique group of eukaryotic plant pathogens that exhibit fungal-like filamentous growth but share little taxonomic affinity to fungi. The characterized gene, pipg1, was identified from the potato late-blight pathogen, Phytophthora infestans, and was predicted to encode a secreted glycoprotein with all the signature sequences of endoPGs. Pipg1 was expressed during preinfection and infection stages. Phylogenetic analysis of endoPGs indicated that pipg1 forms a unique class that is significantly more similar to fungal endoPGs than to plant or bacterial ones. This unexpected affinity between PIPG1 and fungal endoPGs contrasts with phylogenies obtained using ribosomal sequences or compiled protein sequences from mitochondrial and chromosomal genes, raising interesting questions about the evolution of these enzymes in oomycetes. PMID- 11919678 TI - The genome of Plasmodium falciparum encodes an active delta-aminolevulinic acid dehydratase. AB - The enzyme delta-aminolevulinic acid dehydratase (ALAD) catalyses the second reaction in the heme biosynthetic pathway. It has been suggested previously that the malaria parasite Plasmodium falciparum imports this enzyme from the host cell for de novo heme biosynthesis. However, the parasite's genome encodes an orthologue for ALAD. Here we report molecular cloning of a complete cDNA for the parasite's intrinsic ALAD and show it rescues an ALAD-null mutant of Escherichia coli, indicating that the malarial gene encodes a functional ALAD. The malarial ALAD has a long bipartite extension at its N-terminus, which may function as a plastid targeting signal. The amino acid sequence of the enzyme is related most closely to those of plant/algal chloroplast ALADs, though the malarial version may lack the allosteric Mg2+-binding site, which is conserved among chloroplast ALADs. PMID- 11919679 TI - Structure of nuclear-localized cox3 genes in Chlamydomonas reinhardtii and in its colorless close relative Polytomella sp. AB - Several chlorophyte algae do not have the cox3 gene, encoding subunit III of cytochrome c oxidase, in their mitochondrial genomes. The cox3 gene is nuclear encoded in the photosynthetic alga Chlamydomonas reinhardtii and in the colorless alga Polytomella sp. In this work, the genomic sequences of the cox3 genes of these two closely related algae are reported. The cox3 genes of both C. reinhardtii and Polytomella sp. contain four introns in the region encoding the putative mitochondrial-targeting sequences. These four introns show low sequence identities, but their locations are conserved between these species. The cox3 gene of C. reinhardtii has five additional introns in the region encoding the mature subunit III of cytochrome c oxidase. Sequence analysis of intron 6 of the cox3 gene of C. reinhardtii revealed similarity with two sequence elements present in introns of several other nuclear genes from this green alga. In the majority of the genes, these conserved sequences are located either near the 3' end or near the 5' end of the introns. Based on these data, we propose that the colorless genus Polytomella separated from C. reinhardtii after the cox3 gene was transferred to the nucleus. The data also support the evolutionary hypothesis of a recent acquisition of introns in C. reinhardtii. PMID- 11919680 TI - The gene encoding Arabidopsis thaliana mitochondrial ribosomal protein S13 is a recent duplication of the gene encoding plastid S13. AB - A gene encoding mitochondrial S13 is generally present in the mitochondrial genome of higher plants, but is lacking from the Arabidopsis thaliana mitochondrial genome. Previous research has failed to identify a nuclear gene capable of encoding a mitochondrial S13 protein or the protein itself. Doubts have even been raised as to whether a mitochondrial S13 exists in Arabidopsis. Here, we show that the nuclear gene encoding the plastid S13 has been partially duplicated in A. thaliana, such that the copy has lost the exon encoding the plastid transit peptide and acquired a sequence capable of encoding a mitochondrial targeting sequence. The two S13 sequences were fused to green fluorescent protein and shown to be targeted to plastids and mitochondria respectively. PMID- 11919681 TI - A toolbox for a small mammal. PMID- 11919682 TI - Parental genetic contributions in the AXB and BXA recombinant inbred mouse strains. AB - Recombinant inbred (RI) strains are a valuable tool in mouse genetics to rapidly map the location of a new locus. Because RI strains have been typed for hundreds of genetic markers, the genotypes of individual strains within an RI set can be examined to identify specific strain(s) containing the desired region(s) of interest (e.g., one or more quantitative trait loci, QTLs) for subsequent phenotype testing. Specific RI strains might also be identified for use as progenitors in the construction of consomic (chromosome substitution strains or CSSs) or congenic lines or for use in the RI strain test (RIST). To quickly identify the genetic contributions of the parental A/J (A) and C57BL/6J (B) strains, we have generated chromosome maps for each commercially available AXB and BXA RI strain, in which the genetic loci are color-coded to signify the parent of origin. To further assist in strain selection for further breeding schemes, the percentages of A and B parental contributions were calculated, based on the total number of typed markers in the database for each strain. With these data, one can rapidly select the RI strain(s) carrying the desired donor and recipient strain region(s). Because points of recombination are known, starting with RI mice to generate CSSs or congenic lines immediately reduces genomewide screening to those donor-strain regions not already homozygous in the recipient strain. Two examples are presented to demonstrate potential uses of the generated chromosome maps: to select RI strains to construct congenic lines and to perform an RIST for Aliq1, a QTL linked to ozone-induced acute lung injury survival. PMID- 11919683 TI - The KLHL1-antisense transcript ( KLHL1AS) is evolutionarily conserved. AB - Spinocerebellar ataxia type 8 (SCA8) is caused by a CTG expansion in an untranslated, endogenous antisense RNA that overlaps the Kelch-like 1 ( KLHL1) gene. The normal function of this transcript is currently unknown. We have now identified the promoter region for the KLHL1-antisense ( KLHL1AS) RNA and report that a Klhl1as transcript is present in the mouse as well. Human and mouse KLHL1AS are transcribed from homologous promoter regions in the first intron of KLHL1 and extend through the transcription and translation start sites as well as the first splice donor sequence of KLHL1. We found that the mouse Klhl1as RNA is not spliced and terminates in a polyadenylation site in the Klhl1 promoter region, whereas both the present and previous work show that human KLHL1AS is highly variably spliced into processed transcripts that contain up to six exons. Mouse Klhl1as transcript was detected in RNA isolated from the cerebellum and from total adult brain and total fetal tissue, and at a low level in testis and ovary. Similarly, human KLHL1AS is expressed in various brain tissues, including the cerebellum, the tissue most affected by SCA8, and was detected at low levels in testis and kidney. The evolutionary conservation of this antisense/sense transcriptional organization strongly indicates that KLHL1AS transcripts play a significant biological role in both human and mouse, presumably as a regulator of KLHL1 expression. PMID- 11919684 TI - ENU mutagenesis reveals a highly mutable locus on mouse Chromosome 4 that affects ear morphogenesis. AB - Chemical mutagenesis followed by screening for abnormal phenotypes in the mouse holds much promise as a method for revealing gene function. This method is particularly well-suited for discovering genes involved in hearing or balance function, as these defects are relatively easy to screen for in the mouse. We report here the inner ear abnormalities and genetic localization of seven new dominant mutations created by ENU mutagenesis. All seven mutant stocks were identified because of circling and/or head-weaving behavior, which is an indication of balance dysfunction. Investigation of the inner ears of the seven mutant stocks revealed very similar lateral and posterior semicircular canal defects. Studies of the development of the canals in one mutant stock revealed that the affected canals showed reduced outgrowth and delayed canal fusion. Physiological studies performed in one mutant stock showed raised average compound-action-potential thresholds of approximately 10-20 dB sound pressure level (SPL) (depending on frequency), indicating a mild hearing impairment, although scanning electron microscopy performed in several of the mutant stocks revealed no obvious structural defects in the organ of Corti. All seven mutations mapped to the proximal portion of Chromosome (Chr) 4, near the centromere. On the basis of their similar phenotype and map location, we suggest that the seven mutant genes may be allelic and represent a highly mutable locus on Chr 4 that may be particularly susceptible to ENU-induced mutation on the BALB/c genetic background. PMID- 11919685 TI - Y-chromosomal factor is involved in neonatal lethality in (female symbolDDD x male symbolDH- Dh/+) F(1)- Dh/+ male mice. AB - The dominant hemimelia ( Dh) mutation causes various developmental abnormalities in mice. Most F(1)- Dh/+ males, crosses between DDD females and DH- Dh/+ males, have lethal abnormalities during the neonatal period. This is a consequence of synergism among three independent gene loci; that is, the Dh allele on chromosome (Chr) 1, the DDD allele on an X Chr-linked locus, and a Y Chr-linked locus in some strains. With regard to the Y Chr derived from Mus musculus musculus ( M. m. musculus), the Y Chrs of C57BL/6J and BALB/cA caused lethality, but the Y Chr of C3H/HeJ did not, suggesting that not all M. m. musculus Y Chrs are the same. In the present study, whether Y Chrs derived from M. m. domesticus and M. m. castaneus could cause lethality was investigated. Among seven inbred strains, including AKR/J, DDD, RF/J, SJL/J, SWR/J, TIRANO/Ei, and CAST/Ei, Y Chrs of AKR/J, DDD, SJL/J, SWR/J, and TIRANO/Ei caused lethality, but Y Chrs of RF/J and CAST/Ei did not. It was unlikely that the mitochondrial genome of the DDD strain contributed to the lethality. The X Chr-linked locus could not compensate for the role of the Y Chr-linked locus. These results suggest that not all M. m. domesticus Y Chrs are the same. PMID- 11919686 TI - Congenic mapping of a blood pressure QTL on chromosome 16 of Dahl rats. AB - A Chromosome (Chr) 16 segment of the Dahl salt-sensitive (S) rat was shown by linkage to contain a blood pressure (BP) quantitative trait locus (QTL). To verify and further narrow down the region harboring the QTL, we made two congenic strains by replacing two segments of the S rats with the homologous segments of the Lewis (LEW) rats. The construction of these congenic strains was facilitated by a genome-wide marker screening. The two congenic strains contained a segment in common, and BPs of both were significantly lower than that of the S strain. Consequently, a BP QTL could be localized to the overlapping region of about 49.4 centiRay (cR) including the telomere on a radiation hybrid map. Heart weights, left and right ventricular weights, kidney weights, and aortic weights over length were all significantly decreased in the congenic strains compared with the S strain. Thus, there appeared to exist an association between the effects of the QTL on BP and on cardiac, renal, and vascular hypertrophy. PMID- 11919687 TI - Black rat ( Rattus rattus) genomic variability characterized by chromosome painting. AB - Black rats are of outstanding interest in parasitology and infective disease analysis. We used chromosome paints from both the mouse ( Mus musculus) and the Norway rat ( Rattus norvegicus) to characterize the genome of two Black rat subspecies from Italy. Both subspecies have two large metacentrics (n. 1, 4) not present in the Norway rat (2n = 42). Rattus rattus rattus has a diploid number of 2n = 38, while Rattus rattus frugivorous has two small metacentric "supernumerary" or B chromosomes for a diploid number of 2n = 38 + 2B. The 21 mouse paints gave 38 signals on the R. r. rattus karyotype and 39 signals in the R. r. frugivorous karyotype. The two metacentrics, not present in R. norvegicus, were hybridized by mouse 16/1/17 and mouse 4/10/15. These chromosomes are homologous to: RRA1 = RNO 5/7, and RRA4 = RNO 9/11 and not "4/7" and "11/12" as previously reported. Furthermore, the synteny of Chr 13 of the R. r. frugivorous with R. norvegicus Chr 16 and mouse Chrs 8/14 is not complete, because there is a small pericentromeric insertion of RNO Chr 18 (mouse Chr 18). If we consider only the two metacentrics, RRA1 and RRA4, the principal differences between R. norvegicus and R. rattus, then we can propose the derived synteny of 124 genes in the black rat. A comparison of the Z index between rats and mice shows an acceleration of genomic evolution among genus, species, and subspecies. The chromosomal differences between R. r. rattus x R. r. frugivorous suggest that they may be classified as different species because hybrids would produce 50% unbalanced gametes. PMID- 11919688 TI - Genome-wide search for markers associated with bovine spongiform encephalopathy. AB - A genome-wide search for markers associated with BSE incidence was performed by using Transmission-Disequilibrium Tests (TDTs). Significant segregation distortion, i.e., unequal transmission probabilities of alleles within a locus, was found for three marker loci on Chromosomes (Chrs) 5, 10, and 20. Although TDTs are robust to false associations owing to hidden population substructures, it cannot distinguish segregation distortion caused by a true association between a marker and bovine spongiform encephalopathy (BSE) from a population-wide distortion. An interaction test and a segregation distortion analysis in half-sib controls were used to disentangle these two alternative hypotheses. None of the markers showed any significant interaction between allele transmission rates and disease status, and only the marker on Chr 10 showed a significant segregation distortion in control individuals. Nevertheless, the control group may have been a mixture of resistant and susceptible but unchallenged individuals. When new genotypes were generated in the vicinity of these three markers, evidence for an association with BSE was confirmed for the locus on Chr 5. PMID- 11919689 TI - Early embryonic lethality in mice deficient in the p110beta catalytic subunit of PI 3-kinase. PMID- 11919690 TI - Modulation of calcium-activated potassium channels. AB - Potassium currents play a critical role in action potential repolarization, setting of the resting membrane potential, control of neuronal firing rates, and regulation of neurotransmitter release. The diversity of the potassium channels that generate these currents is nothing less than staggering. This diversity is generated by multiple genes (as many as 100 and perhaps more in some creatures) encoding the pore-forming channel alpha subunits, alternative splicing of channel gene transcripts, formation of heteromultimeric channels, participation of auxiliary (non-pore-forming) beta and other subunits, and modulation of channel properties by post-translational modifications and other mechanisms. Prominent among the potassium channels are several families of calcium activated potassium channels, which are highly selective for potassium ions as their charge carrier, and require intracellular calcium for channel gating. The modulation of one of these families, that of the large conductance calcium activated and voltage dependent potassium channels, has been especially widely studied. In this review we discuss a few selected examples of the modulation of these channels, to illustrate some of the molecular mechanisms and physiological consequences of ion channel modulation. PMID- 11919691 TI - Orienting responses and vocalizations produced by microstimulation in the superior colliculus of the echolocating bat, Eptesicus fuscus. AB - An echolocating bat actively controls the spatial acoustic information that drives its behavior by directing its head and ears and by modulating the spectro temporal structure of its outgoing sonar emissions. The superior colliculus may function in the coordination of these orienting components of the bat's echolocation system. To test this hypothesis, chemical and electrical microstimulation experiments were carried out in the superior colliculus of the echolocating bat, Eptesicus fuscus, a species that uses frequency modulated sonar signals. Microstimulation elicited pinna and head movements, similar to those reported in other vertebrate species, and the direction of the evoked behaviors corresponded to the site of stimulation, yielding a map of orienting movements in the superior colliculus. Microstimulation of the bat superior colliculus also elicited sonar vocalizations, a motor behavior specific to the bat's acoustic orientation by echolocation. Electrical stimulation of the adjacent periaqueductal gray, shown to be involved in vocal production in other mammalian species, elicited vocal signals resembling acoustic communication calls of E. fuscus. The control of vocal signals in the bat is an integral part of its acoustic orienting system, and our findings suggest that the superior colliculus supports diverse and species-relevant sensorimotor behaviors, including those used for echolocation. PMID- 11919692 TI - Responses of optokinetic neurons in the pretectum and accessory optic system of the pigeon to large-field plaids. AB - The accessory optic system and pretectum are highly conserved brainstem visual pathways that process the visual consequences of self-motion (i.e. optic flow) and generate the optokinetic response. Neurons in these nuclei have very large receptive fields in the contalateral eye, and exhibit direction-selectivity to large-field moving stimuli. Previous research on visual motion pathways in the geniculostriate system has employed "plaids" composed of two non-parallel sine wave gratings to investigate the visual system's ability to detect the global direction of pattern motion as opposed to the direction of motion of the components within the plaids. In this study, using standard extracellular techniques, we recorded the responses of 47 neurons in the nucleus of the basal optic root of the accessory optic system and 49 cells in the pretectal nucleus lentiformis mesencephali of pigeons to large-field gratings and plaids. We found that most neurons were classified as pattern-selective (41-49%) whereas fewer were classified as component-selective (8-17%). There were no striking differences between nucleus of the basal optic root and lentiformis mesencephali neurons in this regard. These data indicate that most of the input to the optokinetic system is orientation-insensitive but a small proportion is orientation-selective. The implications for the connectivity of the motion processing system are discussed. PMID- 11919693 TI - Encoding of host and non-host plant odours by receptor neurones in the eucalyptus woodborer, Phoracantha semipunctata (Coleoptera: Cerambycidae). AB - In the eucalyptus woodborer, Phoracantha semipunctata, responses from single receptor neurones to host ( Eucalyptus globulus) and non-host ( Pinus pinaster, Olea europeae) plant odours were examined, using gas chromatography linked with electrophysiological recordings. Thirty-two compounds elicited responses that appeared as increased firing rates correlating with the elution of the active components. A subset of neurones classified as type A ( n=17) responded to one compound, a second subset B ( n=15) responded to two compounds and a third subset C ( n=9) responded to three or more compounds. For each neurone of type B or C, the active chemicals were structurally related. Neurones responding to monoterpenes, sesquiterpenes and non-terpenoids, respectively, showed non overlapping response spectra, whereas neurones within one chemical group showed some overlap. The results suggest that most neurones, which receive information about plant odours, are narrowly rather than broadly tuned, each specialised for the detection of one or two related compounds. Although most neurones responded to compounds released by host and non-host species, a few responded specifically to compounds of only one species. This suggests a chemical "fingerprint" mechanism for discrimination between odour blends of the different plants, where only a few compounds may be specific for a particular species or individual. PMID- 11919694 TI - The effects of the visual environment on responses to colour by domestic chicks. AB - It is well known that development of vision is affected by experience, but there are few studies of environmental effects on colour vision. Natural scenes contain predominantly a restricted range of reflectance spectra, so such effects might be important, perhaps biasing visual mechanisms towards common colours. We investigated how the visual environment affects colour preferences of domestic chicks ( Gallus gallus), by training week-old birds to select small food containers distinguished from an achromatic alternative either by an orange or by a greenish-blue colour. Chicks that had been raised in control conditions, with long-wavelength-dominated reflectance spectra, responded more readily to orange than to blue. This was not due to avoidance of blue, as increasing saturation enhanced the chicks' preference for the same hue. The advantage of orange was, however, reduced or abolished for chicks raised in an environment dominated by blue objects. This indicates that responses to coloured food are affected by experience of non-food objects. If colours of ordinary objects in the environment do influence responses to specialised visual signals this might help explain why biological signals directed at birds are often coloured yellow, orange or red; long-wavelength-dominated spectra being more prevalent than short-wavelength dominated spectra. PMID- 11919695 TI - Circadian organization in ruin lizards: phase response curve for melatonin changes with season. AB - Single biweekly injections of exogenous melatonin were given to ruin lizards ( Podarcis sicula) whose circadian locomotor rhythms were recorded under constant temperature (29 degrees C) and darkness. To generate a phase-response curve the direction and amount of phase shift were plotted versus the time at which the injection was given. The results showed, for the second time in the animal kingdom, the existence of a phase-response curve for melatonin. The area under the delay part of the phase-response curve is much greater than the area under the advance part, and there is no dead zone. The phase-response curve for melatonin was generated in summer, but the repetition of experiments in autumn winter did not generate any phase-response curve for melatonin in these seasons. In autumn-winter no one phase of the ruin lizards' circadian cycle was shifted in response to melatonin injections. This confirms several previous results in ruin lizards, showing that melatonin is centrally involved in determining circadian organization in summer and only marginally involved in autumn-winter. PMID- 11919696 TI - The effect of preceding sonar emission on temporal integration in the bat, Megaderma lyra. AB - The present study investigated whether and to which extent temporal integration in bats is influenced by echolocation behavior. One way to quantify temporal integration is to measure the detection threshold for a pair of short tone pips as a function of the temporal separation between the pips. To asses the effect of preceding sonar emission on temporal integration in the bat, Megaderma lyra, the detection thresholds of identical subjects were measured in a passive as well as in an active paradigm. In the passive paradigm, the presentation of the pip pairs was independent of the bats' sonar emissions; in the active paradigm, the presentation was triggered by the bats' sonar emissions. In both cases, the bats showed a very short integration time in the range of 100-200 micros. Moreover, the comparison of the active and passive results within each bat revealed no systematic differences in the two measuring paradigms. These results indicate that temporal integration is not influenced by echolocation. Simulations with a computer model of cochlear filtering based on measurements of M. lyra cochlear tuning suggest that the perceptual temporal integration is dominated by the integration of the cochlear filters. PMID- 11919697 TI - Behavioral titration of a magnetic map coordinate. AB - Spatial variation in the inclination of the geomagnetic field has been implicated in the map component of homing by eastern red-spotted newts Notophthalmus viridescens. Here we show that when newts are exposed to small changes in magnetic inclination, the most dramatic effects on homing orientation occur at values close to the 'home value', as predicted by the magnetic map hypothesis (Phillips 1996). Newts reverse the direction of homing orientation over a range of inclination of 0.5 degrees spanning the home value, providing further evidence that magnetic inclination or one of its components (i.e., vertical or horizontal intensity) is used to derive map information. PMID- 11919698 TI - "Allohormones": a new class of bioactive substances or old wine in new skins? PMID- 11919700 TI - Properties of Na(+)-dependent nucleoside transport in the proximal and distal small intestine of cows. AB - Large amounts of nucleic acids associated with rumen microorganisms are digested in the proximal part of the small intestine of ruminants. We studied how the proximal-distal gradient in nucleic acid digestion is related to activity of Na(+)-nucleoside transporters in brush border membrane vesicles isolated from the proximal and distal small intestine of cows. Two Na(+)-dependent nucleoside transporters with overlapping substrate specificity were shown to be present at the two intestinal sites, one for pyrimidine nucleosides and one for purine nucleosides. Affinity constants (K(m)-values) for both thymidine and guanosine transport were similar at the two intestinal sites, while transport capacity (V(max)) was 2-3 times higher in the proximal than in the distal small intestine. Glucose and alpha-methyl-D-glucoside (0.1 mmol/l or 2 mmol/l) inhibited transport of thymidine and guanosine markedly only in the proximal small intestine. It is concluded that absorption of nucleosides by the two Na(+)-nucleoside transporters reflects the proximal-distal gradient in nucleic acid digestion. PMID- 11919701 TI - Seasonal changes in morphology and function of the gastrointestinal tract of free living alpine marmots ( Marmota marmota). AB - The gastrointestinal tracts of 76 free-living alpine marmots ( Marmota marmota) shot during a population control program in Switzerland were collected and analysed for patterns of change in morphology and function over the period from emergence from hibernation in April to just before re-entry into hibernation in September. Between first emergence and mid-summer (July) the fresh tissue mass of the stomach increased by 105%, the small intestine by 259% (among the largest recorded for a mammal), caecum by 185%, proximal colon by 138%, and distal colon by 144%. Mitotic activity was greatest in the small intestine; the mitotic index was high (40%) compared with indexes in the stomach and hindgut (approximately 4%) even at emergence, and increased to approximately 60% by mid-summer. Microbial activity in the caecum was also significant at emergence. The stomach (length) and caecum (length and fresh mass) increased in response to ingested food earlier than did the small intestine. Between mid-summer and September there were decreases in small intestinal tissue mass and mitotic activity. It is concluded that the gastrointestinal tract of alpine marmots probably continues to function throughout hibernation at a low level, with a mid-winter trough as part of an endogenous circannual rhythm. However, after emergence in spring, increases in size and activity of the tract appear to be a response to ingested food rather than to an endogenous signal. The early signs of down-regulation of the small intestine before re-entry into hibernation, together with its delayed up regulation in response to food in spring, are consistent with the high costs of maintaining this section of the digestive system. PMID- 11919702 TI - The effects of perfusion of the cutaneous vasculature on sodium uptake across isolated frog skin. AB - Perfusion of cutaneous capillaries in isolated frog skin may remove an unstirred layer along the basolateral membrane of the epidermis that may affect the rate of cutaneous Na(+) uptake. To test this hypothesis, the cutaneous artery and vein of a bullfrog were cannulated to allow perfusion of isolated flank skin while the rate of Na(+) influx was determined. Rates of sodium influx with and without perfusion were determined in the same experiment. Na(+) uptake increased by 59+/ 4.8% during the 1st 0.5 h of perfusion relative to the control, pre-perfusion period and then remained at 26+/-5.3% above control values. Concomitant with the increase in sodium uptake, the transepithelial potential difference fell by ca. 10% within the 1st 0.5 h of perfusion. The amount of labeled sodium leaving the skin in the venous effluent decreased exponentially in the 1st 0.5 h of perfusion, suggesting a wash out of an unstirred layer within the interstitial fluid. Sodium in the venous outflow accounted for ca. 25% of the sodium uptake during each perfusion period. Perfusion of the cutaneous vasculature thus has a significant effect on Na(+) transport and may potentially play a role in the acute regulation of cutaneous ion transport. PMID- 11919704 TI - Modulation of the cardiac pacemaker of Drosophila: cellular mechanisms. AB - The myogenic cardiac pacemaker of Drosophila melanogaster responds to a range of neurotransmitters and hormones by adjusting heart rate. These cardioactive substances ultimately affect the activity of ion channels comprising the pacemaker. We report here work utilizing genetic variants and pharmacological tools to explore a subset of possible mechanisms for this cellular signaling, specifically: receptors, cAMP, cGMP, G proteins, and calcium. We found that alpha(1) adrenergic and 5-hydroxytryptamine(2) (5-HT(2)) receptors are critical components of mediating modulation of heart rate. There was no evidence that the cAMP system is part of the modulatory mechanism. cGMP is likely to be integral to one active pathway, as non-hydrolyzable forms of this cyclic nucleotide increase heart rate, and flies bearing the mutation sitter, a recessive allele of the foraging gene, which encodes a cGMP-dependent kinase, have tachycardia. Heart rhythm is affected by pertussis toxin and by agonists and antagonists of both alpha(1) adrenergic and 5-HT(2) receptors; this suggests involvement of two different types of G proteins. The l(4)16/ciD line, containing a mutation in CaM kinase II, eliminates pacemaker responsiveness to serotonin but is without effect on norepinephrine sensitivity. This result is the same as that for the CaM kinase II enzyme inhibitor KN-93. This work establishes a framework for further investigations into the control of the cardiac pacemaker, and expands the applicability of the Drosophila heart model. PMID- 11919703 TI - Effects of adenosine on the contractility of normoxic rainbow trout heart. AB - Temperature strongly affects oxygen solubility in water, oxygen convection in the blood and locomotor activity of the fish. Since oxygen supply and demand are temperature dependent, it was hypothesized that the purinergic control of the heart, one of the most important mediators in oxygen-limited conditions, might also show temperature dependence. Therefore, the present study examines the effects of adenosine (Ado), a purinergic agonist, on the contractile and electrical activity of the thermally acclimated trout ( Oncorhynchus mykiss Walbaum) heart. The fish were acclimated to either 4 degrees C or 17 degrees C and the experiments were conducted at the acclimation temperatures of the animals. In spontaneously beating hearts, Ado had a negative chronotropic and a positive inotropic effect in warm-acclimated rainbow trout while no response was detected in cold-acclimated trout. In paced atrial and ventricular preparations, Ado had a negative inotropic effect in both warm- and cold-acclimated fish, and the response was strongest in the atria of warm-acclimated trout. Ado shortened the duration of contraction 12-14% in atrial preparations but had no effect in ventricular muscle. Ado (10(-4) mol l(-1)) increased the density of the inwardly rectifying K(+) current from -3.5+/-0.6 pA pF(-1) to -8.4+/-1.4 pA pF(-1) (at 120 mV) in atrial myocytes of warm-acclimated trout but was without effect in atrial myocytes of cold-acclimated trout (-2.4+/-0.8 pA pF(-1) vs. -2.1+/-0.9 pA pF(-1)). Ado had no effect on K(+) currents of ventricular cells in either acclimation group. These results indicate that the effects of Ado on cardiac contractility and electrical activity are stronger in warm-acclimated than in cold-acclimated trout when measured at the physiological body temperatures of the fish. The balance between oxygen demand and supply of the heart might be better in the cold where more environmental oxygen is available and the power of the muscles is weaker thereby reducing the need for the purinergic control of the heart. Temperature-dependence of Ado response in the trout heart warrants that temperature should be taken into consideration when the purinergic system of the ectotherms is studied. PMID- 11919705 TI - The nature of the acetylcholine and 5-hydroxytryptamine receptors in buccal smooth muscle of the pest slug Deroceras reticulatum. AB - The characteristics of the acetylcholine (ACh) and 5-hydroxytryptamine (5-HT) receptors of Deroceras buccal muscle were examined using specific pharmacological probes and sucrose gap electrophysiological analysis. ACh induced concentration dependent smooth tonic contractures coupled with considerable depolarisation from the normal resting membrane potential of -30.6 mV. The use of choline ester analogues such as carbachol, propionylcholine and butyrylcholine, specific cholinergic agonists such as nicotine, muscarine, bethanecol and pilocarpine and antagonists such as d-tubocurarine, succinylcholine, hexamethomium, atropine, gallamine, pirenzepine and scopolamine indicated that the ACh receptor showed both nicotinic and muscarinic characteristics; the muscarinic activity resembled that of a mammalian M(2)-like receptor. Alternatively, it can not be ruled out that both mammalian types of receptor may be present in this preparation since both nicotine and muscarine induced noticeable tension. 5-HT application induced characteristic dose-dependent phasic contractions accompanied by small but quite consistent depolarisations. Serotonergic agonist and antagonist experiments using 1-(3-chlorophenyl) piperazine, 1-(m-chlorophenyl) biguanide, methiothepin, methysergide and metoclopramide strongly suggested that the 5-HT receptor showed closest pharmacological affinity with the 5-HT(1) receptor class of mammals but with some 5-HT(2) activity. In view of the phylogenetic gap between molluscs and mammals it is not surprising that the ACh and 5-HT receptors of Deroceras can not be properly classified by conventional mammalian terminology. PMID- 11919706 TI - Influence of hyperosmotic shrinkage and beta-adrenergic stimulation on red blood cell volume regulation and oxygen binding properties in rainbow trout and carp. AB - Whole blood from rainbow trout and carp was subjected to hyperosmotic shock and subsequent beta-adrenergic stimulation (isoprenaline) at different oxygen tension ( PO(2)) and carbon dioxide tension ( PCO(2)) levels with the aim to evaluate changes in red blood cell (RBC) volume, pH and ion concentrations and their ultimate effect on blood O(2) transport characteristics. Hyperosmolality (addition of NaCl) induced RBC shrinkage, which was followed by a regulatory volume increase (RVI) that was larger at low than at high PO(2)and more complete in carp than in trout. Carp RBC showed practically full volume recovery within 140 min at low PO(2)and partial recovery at high PO(2), whereas RVI was partial under all PO(2)and PCO(2)conditions in trout. The RVI increased intracellular [Na(+)], water content, and, in carp, also pH (pHi), suggesting activation of Na(+)/H(+) exchange. In trout RBCs, activation of RVI was rapid but succeeded by deactivation. In carp RBCs, activation of Na(+) influx was slower but it continued, allowing full volume recovery. Shrinkage of the RBCs was associated with only minor decreases in blood oxygen saturation and oxygen affinity in both species. Thus, the oxygen affinity decrease expected on the basis of the increased concentration of intracellular haemoglobin and organic phosphates was small, and it appeared to some extent countered during RVI by an oxygen affinity increase via increased pHi. Addition of isoprenaline increased RBC volume and pHi and increased Hb oxygen saturation. The beta-adrenergic response was stronger at low compared to high PO(2) and at high compared to low PCO(2). The PO(2) dependency was largest in carp, whereas the PCO(2) (pH) dependency was more expressed in trout. The adrenergic response of trout RBCs was similar under isoosmotic and hyperosmotic conditions. In carp RBCs, the response was absent at high PO(2) under isoosmotic conditions, but interestingly it could be induced under hyperosmotic conditions. The data suggest that the RBC shrinkage occurring in fish moving from freshwater to seawater has minimal impact on blood O(2) binding properties. PMID- 11919707 TI - Carbon isotope ratios in exhaled CO(2) can be used to determine not just present, but also past diets in birds. AB - We show that an animal's past and present diet can be distinguished through the delta(13)C of exhaled CO(2). The exhaled delta(13)C of 12 pigeons fed solely corn (a C(4) plant) for 30 days was -13.63 per thousand (+/-0.30). We then fed six pigeons wheat (a C(3) plant) and continued to feed the other six corn. After 48 h the exhaled delta(13)C from the corn-fed pigeons was unchanged; that from the wheat-fed pigeons was -20.5 per thousand. We then fasted three of the wheat-fed pigeons for 3 days, after which their exhaled delta(13)C was -14.96 per thousand, while it was -13.57 per thousand in corn-fed pigeons, and -22.22 per thousand in pigeons that continued on wheat. Thus, we could infer diet from the (13)C/(12)C ratios of exhaled CO(2). Significantly, breath samples from fasted pigeons also revealed that they had eaten corn when their lipid stores were formed. We also showed that the change in the (13)C/(12)C of exhaled CO(2) had a half-life of approximately 3.5 h, and a time constant of approximately 6.7 h. Thus one can infer past and present diet from exhaled delta(13)C alone, if the initial breath sample is followed by a fasted breath sample, without harming the animal or having to recapture it successively. PMID- 11919708 TI - Ubiquitin conjugate dynamics in the gut and liver of hibernating ground squirrels. AB - Protein synthesis is severely depressed in hibernating mammals. In the absence of significant protein synthesis, the continued turnover of proteins as a function of normal cellular activity would result in the net depletion of protein pools. We measured levels of ubiquitylated proteins in the gut of thirteen-lined ground squirrels ( Spermophilus tridecemlineatus) and liver of golden-mantled ground squirrels ( Spermophilus lateralis). In both tissues, ubiquitin conjugate concentrations increased during entrance into torpor and were elevated 2-3 fold by late torpor compared with levels in active animals. The data are consistent with a depression of proteolysis with a resultant high level of ubiquitylated proteins during the natural hypothermia of torpor. The periodic returns to euthermy during the hibernation season allow for degradation of these conjugated proteins and may serve to restore protein pools. PMID- 11919710 TI - Tetrad-FISH analysis reveals recombination suppression by interstitial heterochromatin sequences in rye (Secale cereale). AB - Tetrad analysis is a genetic method that can locate genes and centromeres on a linkage map with a high degree of precision. Despite its effectiveness and accuracy, application of this method is generally limited to fungi, algae and mosses. Here we demonstrate a new method of tetrad analysis that is applicable to other organisms. This combines tetrad analysis with fluorescence in situ hybridization (FISH), and is thus referred to as tetrad-FISH analysis. We demonstrate the effectiveness of this method using tetrads of rye, Secale cereale. The rye strain JNK contains interstitial heterochromatin in a region of Chromosome 2R. We have previously cloned the tandemly repeated sequence forming this heterochromatin in the plasmid pScJNK. We performed FISH using pScJNK as the probe on tetrads obtained from heterozygotes for the heterochromatin region. The frequency of tetrads demonstrating positive signals in two cells that are diagonally opposite one another must correspond to the frequency of recombination in the interval between the heterochromatin and the centromere. Comparison between the results of tetrad-FISH analysis and linkage maps based on RFLP markers clearly indicated that heterochromatin strongly suppresses recombination of whole chromosomal regions. We discuss the effectiveness of tetrad-FISH analysis, particularly for the localization of functional centromeres in linkage maps. PMID- 11919709 TI - Characterization of the kynurenine 3-monooxygenase gene corresponding to the white egg 1 mutant in the silkworm Bombyx mori. AB - Kynurenine 3-monooxygenase (KMO, EC 1.14.13.9), which catalyzes the oxidation of kynurenine to 3-hydroxykynurenine, is involved in the synthesis of ommochrome pigments in insects. A silkworm mutant, white egg 1 ( w-1), has been shown to be deficient in this enzyme activity. The mutant is characterized morphologically by its white eyes and the fact that the females lay white eggs. To analyze the relationship between the KMO gene and the mutation, we first determined the entire sequence of a full-length 2.0-kb cDNA and examined its expression pattern in the wild type. The cDNA sequence contains one ORF encoding a polypeptide of 456 amino acids, and transcripts were detected in the larval Malpighian tubules and the pupal ovaries, but not in other tissues. Southern analysis and nucleotide sequencing showed that the KMO gene is present in a single copy and consists of ten exons distributed over a 16-kb region. Comparison of the transcripts between the wild type and mutant silkworms showed that the wild type expressed a single transcript, whereas the mutant exhibited markedly reduced amounts of two transcripts with sizes of 2.0 kb and 1.8 kb. Nucleotide sequence analysis of these mutant transcripts indicated that sequences corresponding to the ninth and tenth exons were missing. Inverse PCR and Southern analysis of the mutant gene demonstrated that the corresponding genomic region was deleted in the w-1 mutant. PMID- 11919711 TI - The tobacco bZIP transcription factor BZI-1 binds to G-box elements in the promoters of phenylpropanoid pathway genes in vitro, but it is not involved in their regulation in vivo. AB - Screening of a tobacco (Nicotiana tabacum) cDNA library resulted in the isolation of a clone encoding the bZIP transcription factor BZI-1. With respect to amino acid sequence, conservation of protein domains, genomic exon-intron structure and expression pattern, BZI-1 is closely related to CPRF2, OHP1/2, BLZ1 and REB, a group of bZIP proteins which have been described in a number of dicot and monocot species. BZI-1 exhibits the characteristics of a transcription factor. It binds to G-box and C-box cis-elements in vitro, it is localised in the nucleus, and the N-terminal region of BZI-1 functions as an activation domain in both yeast and plant cells. Since BZI-1-related transcription factors have been isolated from dicots by in vitro binding to G-box elements in the chalcone synthase ( CHS) promoter, it has been suggested that phenylpropanoid pathway genes, such as CHS and PAL (phenylalanine ammonia-lyase), are targets of these proteins in vivo. However, after infection with Pseudomonas syringae or Tobacco Mosaic Virus, no changes in pathogen-induced PAL expression were observed in transgenic plants expressing increased levels of BZI-1 or a dominant negative form of the protein, BZI-1-DeltaN. In contrast to the tissue-specific expression of CHS and PAL, BZI-1 was found to be ubiquitously expressed in tobacco plants. Furthermore, no changes in the tissue-specific expression of PAL or CHS were observed in plants that were transgenic for BZI-1-DeltaN. Expression of a VP16-BZI-1 fusion protein would be expected to result in constitutive activation of the BZI-1 target genes. However, tetracycline-dependent expression of a VP16-BZI-1 protein in tobacco plants did not result in activation of CHS or PAL. On the basis of these data, we conclude that the phenylpropanoid pathway genes analysed are not targets of BZI-1 in vivo. Thus, the pattern of in vitro DNA binding of transcription factors need not always reflect their in vivo function. PMID- 11919713 TI - Organization and activation of the late promoters of phiCTX, a cytotoxin converting phage from Pseudomonas aeruginosa. AB - The late genes of the temperate phage of Pseudomonas aeruginosa are organized in an analogous fashion to the corresponding transcription units of the Escherichia coli P2 and P2-like phages. Sequence analysis of four putative late promoter regions, PP(phiCTX), PO(phiCTX), PV(phiCTX) and PF(phiCTX), reveals no similarity to sigma(70)-type promoters or promoter consensus sequences found in Pseudomonas, indicating the apparent need for a phage-encoded protein to control the expression of phiCTX late genes. To elucidate the mode of expression of the late genes, we fused the putative late promoter regions to the promoterless lacZalpha gene, which encodes the N-terminal part of beta-galactosidase as a reporter enzyme, in the promoter-probe vector pME4510. The candidate transactivator gene orf34 was cloned into expression vector pHA10, to generate the plasmid pHA34. The two recombinant plasmids were introduced together into E. coli XL1-Blue and P. aeruginosa PAO1S-Lac. Our results demonstrate that in phiCTX three late promoters (PP(phiCTX), PO(phiCTX), and PF(phiCTX)) are activated upon induction by IPTG in PAO1S-Lac carrying the cloned promoters and pHA34. Deletions and base-pair substitutions obtained by PCR-mediated mutagenesis demonstrated that two conserved sequences, TTGTAG-N(9)-cTACAa and GcCGCGCGCGCGgC, are essential for effective late gene expression. Whereas the late promoters were active in P. aeruginosa, only weak beta-galactosidase activity was obtained in E. coli. PMID- 11919712 TI - Differential regulation of genes transcribed by nucleus-encoded plastid RNA polymerase, and DNA amplification, within ribosome-deficient plastids in stable phenocopies of cereal albino mutants. AB - We isolated stable albino plants of barley and maize by inhibiting plastid protein synthesis with streptomycin and propagating bleached seedlings in the absence of antibiotics in vitro. Albino plants are deficient in plastid translation products and plastid ribosomal RNAs, and are stable phenocopies of the barley albostrians and maize iojap mutants, which contain ribosome-free plastids. Once plastid ribosomes are lost they cannot be re-synthesized because about one-third of plastid ribosomal proteins are themselves plastid encoded. The group II/subgroup IIA intron in plastid rpl2 transcripts was not processed in albinos, providing strong evidence for the absence of plastid translation. Photosynthesis-related plastid mRNAs and plastid tRNAs were down-regulated in albino leaves. A differential influence of plastid ribosome deficiency on mRNA levels allowed us to divide genes transcribed by nucleus-encoded plastid RNA polymerase into two groups. Northern analysis revealed increases in the levels of clpP, rpl2, rpl23, rps15 and rpoB mRNAs in total RNA from albino leaves relative to those in green leaves. In contrast, albinism did not increase the band intensities of rps2 and rps4 messages. Plastid ribosome-associated factor(s) or plastid-encoded product(s) play a role in the initiation, termination, processing or stability of transcripts containing trnG(UCC) and rps4. Excision and 100-fold amplification of a 5.2-kb region of plastid DNA encompassing the trnG(UCC) and trnE(UUC) genes was observed in one of four albino barley plants. Gene amplification was correlated with the accumulation of abundant novel transcripts derived from regions flanking the trnG(UCC) gene. PMID- 11919714 TI - A 70-kDa chloroplast DNA polymerase from pea ( Pisum sativum) that shows high processivity and displays moderate fidelity. AB - A 70-kDa chloroplast (ct) DNA polymerase from pea has been purified to apparent homogeneity. The ct DNA polymerase was insensitive to dideoxynucleotides (d(2) NTP) but showed high sensitivity to phosphonoacetic acid. The enzyme lacked any detectable 5'-->3' exonuclease activity but showed 3'-->5' exonuclease activity. The polymerase displayed high processivity (3 kb) and moderate fidelity, which may be sufficient for the faithful replication of the 140-kb pea ct genome. A 43 kDa accessory protein increased the polymerization rate but did not affect the rate of mis-incorporation in vitro, thus indicating that the domains for polymerisation and proof reading may be spatially separate. PMID- 11919715 TI - The Hopscotch Jak kinase requires the Raf pathway to promote blood cell activation and differentiation in Drosophila. AB - Cytokines regulate the development and differentiated functions of hematopoietic cells by activating multiple signaling pathways, including the Jak-Stat pathway, the PI3-kinase pathway, and the Ras/Raf pathway. While the Jak-Stat interaction has been extensively studied, the relationship between this pathway and other cytokine-induced signaling pathways is not fully understood. In Drosophila melanogaster, mutations that result in hyperactivity of the Jak kinase Hopscotch (Hop) cause an activation of the larval blood cell encapsulation response, including blood cell aggregation and differentiation of plasmatocytes into apparent lamellocytes. Here, we demonstrate that Hop requires the activity of the Raf pathway to promote the activation response of larval plasmatocytes, and provide evidence to suggest that the Hop and D-Raf proteins physically interact. We also show that basal level activity of the Raf pathway is required for the accumulation of circulating blood cells. PMID- 11919716 TI - Most meiotic CAG repeat tract-length alterations in yeast are SPO11 dependent. AB - The expansion of trinucleotide repeat sequences associated with hereditary neurological diseases is believed from earlier studies to be due to errors in DNA replication. However, more recent studies have indicated that recombination may play a significant role in triplet repeat expansion. CAG repeat tracts have been shown to induce double-strand breaks (DSBs) during meiosis in yeast, and DSB formation is dependent on the meiotic recombination machinery. The rate of meiotic instability is several fold higher than mitotic instability. To determine whether DSB repair is responsible for the high rate of repeat tract-length alterations, the frequencies of meiotic repeat-tract instability were compared in wild-type and spo11 mutant strains. In the spo11 background, the rate of meiotic repeat-tract instability remained at the mitotic level, suggesting that meiotic alterations of CAG repeat tracts in yeast occur by the recombination mechanism. Several of these meiotic tract-length alterations are due to DSB repair involving use of the sister chromatid as a template. PMID- 11919717 TI - Evolutionary analysis of S-RNase genes from Rosaceae species. AB - Eight new cDNA sequences for S-RNases were cloned and analysed from almond (Prunus dulcis) cultivars of European origin, and compared to published sequences from other Rosaceae species. Insertions/deletions of 10-20 amino acid residues were detected in the RC4 and C5 domains of S-RNases from almond and sweet cherry. The S-RNases of the Prunus species and those of the genera Malus and Pyrus formed two distinct groups on phylogenetic analysis. Nucleotide substitutions were analysed in the S-RNase genes of these species. The S-genes of almond and sweet cherry have a lower Ka/Ks value than those of apple, pear and wild apple do. The fact that there is no fixed difference between the S-RNase genes of almond and sweet cherry, or between apple and pear, suggests that nucleotide substitutions only introduce transient polymorphism into the two groups, and rarely became fixed and contribute to divergence. Through the comparative study of 17 S-RNase genes from the genus Prunus and 18 from the genera Malus and Pyrus, some fixed nucleotide differences between the two groups were identified. These differences do not appear to be the result of selection for adaptive mutations, since the number of replacement substitutions is not significantly greater than the number of synonymous substitutions. S-RNase genes of almond and sweet cherry, and of apple and pear, showed little heterogeneity in nucleotide substitution rates. However, heterogeneity was observed between the two groups of S-alleles, with the Prunus alleles exhibiting a lower rate of non-synonymous substitutions than alleles from Malus and Pyrus. The evolutionary relationships between these species are discussed. PMID- 11919718 TI - Aberrant transposition of a Tc1-mariner element, impala, in the fungus Fusarium oxysporum. AB - We previously determined that the impalaD transposable element of Fusarium oxysporum was able to mobilize a non autonomous copy of impala ( niaD::imp::hph), inserted in the niaD gene encoding nitrate reductase. Generally, mobilization results in the recovery of Nia(+) revertants at low frequency. In the course of this study, we recovered a transformant that gave rise to Nia(+) revertants at a high rate. These revertants displayed atypical phenotypes and showed a niaD hybridization pattern different from that in more typical revertants. Molecular analysis of the structure of the transformant and atypical revertants indicated that (i) in the transformant, two copies of impala, one defective and one active, were inserted at the same genomic locus in a head-to-head orientation; and (ii) all the revertants analyzed presented the same chromosomal rearrangement, an inversion resulting in the replacement of the niaD promoter by a new sequence containing a cryptic promoter. We also frequently observed additional DNA rearrangements (deletion or inversion) in these revertants. The sequences at the rearrangement junctions indicated the occurrence of a transposition event that used the ITRs (Inverted Terminal Repeats) of separate transposons arranged in direct orientation. These features can be interpreted as the consequences of an aberrant transposition process. Such a process may account for the rearrangements observed in some genomic regions containing multiple transposon ends, and could serve as a mechanism for the generation of genetic diversity. PMID- 11919719 TI - The deubiquitinating enzyme Ubp21p of fission yeast stabilizes a mutant form of protein kinase Prp4p. AB - The protein kinase Prp4p of Schizosaccharomyces pombe is involved in control of the formation of active spliceosomes, phosphorylating the spliceosomal component Prp1p. The kinase domain of Prp4p is closely related to cyclin-dependent kinases (CDKs) and mitogen-activated kinases (MAPKs). A mutational analysis of the highly conserved amino acid sequence ALKHP in subdomain XI of this kinase showed that structural features of this sequence are important for the function of the kinase. We identified ubp21 as a high-copy-number suppressor of a mutation in the ALKHP motif. Characterization of this gene revealed that it encodes a deubiquitinating enzyme belonging to the family of ubiquitin-specific processing proteases (Ubps). The results presented in this report are consistent with the notion that the deubiquitinating activity of Ubp21p may be involved in regulating the steady-state levels of proteins including Prp4p. PMID- 11919720 TI - Functional dissection of Pdr1p, a regulator of multidrug resistance in Saccharomyces cerevisiae. AB - Pleiotropic drug resistance in the yeast Saccharomyces cerevisiae results mainly from the overexpression of genes encoding membrane efflux pumps, the so-called ABC and MFS transporters. These pleiotropic drug resistance loci are under the control of the key transcription factors Pdr1p and Pdr3p. We have identified and characterized several new domains of Pdr1p. By testing a series of LexA-PDR1 derivatives for their capacity to activate a GAL1-lacZ reporter gene we have shown that the C-terminal domain of Pdr1p comprising amino acids 879-1036 is involved in transcriptional activation, and that the point mutation pdr1-8 increases its efficiency. Removal of amino acids 1006-1029, which include a polyasparagine stretch, decreases the activation function. Internal deletions within Pdr1p reveal the presence of a large regulatory domain, and a short but strong inhibitory subdomain spanning amino acids 257-316, in which the up regulating mutations pdr1-2, pdr1-6 and pdr1-7 are located. A mini-Pdr1p consisting of only the DNA-binding and the activation domains strongly up regulates the expression of the major target genes PDR5, SNQ2 and YOR1, resulting in enhanced multidrug resistance. PMID- 11919721 TI - Development of S-SAP markers based on an LTR-like sequence from Medicago sativa L. AB - The Sequence-Specific Amplification Polymorphism (S-SAP) method, recently derived from the Amplified Fragment Length Polymorphism (AFLP) technique, produces amplified fragments containing a retrotransposon LTR sequence at one end and a host restriction site at the other. We report the application of this procedure to the LTR of the Tms1 element from Medicago sativa L. Genomic dot-blot analysis indicated that Tms1 LTRs represent about 0.056% of the M. sativa genome, corresponding to 16 x 10(3) copies per haploid genome. An average of 66 markers were amplified for each primer combination. Overall 49 polymorphic fragments were reliably scored and mapped in a F(1) population obtained by crossing diploid M. falcata with M. coerulea. The utility of the LTR S-SAP markers was higher than that of AFLP or SAMPL (Selective Amplification of Microsatellite Polymorphic Loci) markers. The efficiency index of the LTR S-SAP assay was 28.3, whereas the corresponding values for AFLP and SAMPL markers were 21.1 and 16.7, respectively. The marker index for S-SAP was 13.1, compared to 8.8 for AFLP and 9.5 for SAMPL. Application of the Tms1 LTR-based S-SAP to double-stranded cDNA resulted in a complex banding pattern, demonstrating the presence of Tms1 LTRs within exons. As the technique was successfully applied to other species of the genus Medicago, it should prove suitable for studying genetic diversity within, and relatedness between, alfalfa species. PMID- 11919722 TI - Identification of a chromosomal tra-like region in Agrobacterium tumefaciens. AB - The virD4 gene is one of the virulence genes present on the pTiC58 plasmid of Agrobacterium tumefaciens. Unexpectedly, we found that a pTi-free A. tumefaciens strain carried a protein of similar size to the plasmid-encoded VirD4 protein which reacted with VirD4-specific antibodies. This suggested that this strain may contain a homologue of the VirD4 protein. A chromosomal fragment encoding a protein of similar sequence to VirD4 was isolated and a 7.8 kilobase region surrounding the gene encoding this putative homologue was sequenced. This region contained four open reading frames, encoding putative proteins similar to proteins of known bacterial transfer and conjugation systems, viz., orf1 encoded a putative homologue of the TraA protein of the Rhizobium symbiosis plasmid pNGR234 and the TraA protein encoded by pTiC58 from A. tumefaciens plasmid pTiC58, orf3 encoded a protein very similar to the MobC protein encoded by the IncQ plasmid RSF1010 of E. coli and to MobS encoded by pTF1 from Thiobacillus ferrooxidans, whereas the predicted product of orf4 displayed similarity to the TraG protein encoded by the IncPalpha plasmid RP4 of E. coli, TraG and VirD4 encoded by A. tumefaciens plasmid pTiC58. The product of orf2 showed no significant similarity to any known protein. Preliminary assays with two orf4 mutants suggested that the product of this orf is involved in DNA transfer. The 7.8 kb chromosomal fragment seems to be closely related to the tra region of different conjugative plasmids and appears to be confined to Agrobacterium species, raising the question of the role of a chromosomal tra-like region during evolution. PMID- 11919723 TI - Lactose metabolism and cellulase production in Hypocrea jecorina: the gal7 gene, encoding galactose-1-phosphate uridylyltransferase, is essential for growth on galactose but not for cellulase induction. AB - Lactose is at present the only soluble carbon source which can be used economically for the production by Hypocrea jecorina (= Trichoderma reesei) of cellulases or heterologous proteins under the control of cellulase expression signals. However, the mechanism by which lactose triggers the formation of cellulases is unknown. To enhance our understanding of lactose metabolism and its relationship to cellulase formation, we have cloned and characterized the gal7 gene (for galactose-1-phosphate uridylyltransferase) of H. jecorina. The gene encodes a polypeptide of 43.8 kDa, the sequence of which exhibits a moderate level of identity (about 50%) to that of the Gal7 proteins of Saccharomyces cerevisiae and Kluyveromyces lactis, and contains an active-site signature typical for galactose-1-phosphate uridylyltransferase family 1. H. jecorina gal7 is not clustered with other genes of galactose metabolism. A single 1.7-kb transcript is synthesized constitutively during the rapid growth phase and accumulated to twice this level during incubation in the presence of D-galactose and L-arabinose and the corresponding polyols (dulcitol, arabitol). A gal7 deletion mutant, constructed by replacing the gal7 reading frame by the H. jecorina pyr4 gene, was unable to grow on D-galactose between pH 4.5 and 7.5, thus proving that in H. jecorina gal7 is essential for metabolism of D-galactose, whereas the growth rate of the mutant on lactose was only reduced by about 50%. The rate of formation of cellobiohydrolase Cel7A and the abundance of the corresponding (cbh1) transcript during growth on lactose was only slightly lower in the absence of gal7, but a significant delay in decay of the cbh1 transcript was noted during later stages of growth. The results suggest that H. jecorina uses only the Leloir pathway for metabolism of D-galactose and lactose. Furthermore, we conclude that metabolism of lactose past the galactose-1 phosphate step is not essential for cellulase formation. PMID- 11919724 TI - Antibiotic prophylaxis in patients receiving hematopoietic stem cell transplant. AB - Effective prophylaxis against specific infections has allowed increasingly potent conditioning regimens to be given, thereby prolonging survival in HSCT recipients. The Centers for Disease Control and Prevention, in collaboration with numerous professional societies, has recently published guidelines to codify and advance this approach. Controversy remains in several areas but, curiously, the most intense debate concerns prevention of bacterial infections, the most extensively studied of all of the approaches. Central to this debate are the competing priorities of a potentially ill patient on the one hand vs the long term consequences of unchecked antibiotic use. The emergence in the 1990s of vancomycin-resistant Enterococcus demonstrated all too vividly how devastating such an end result could be. This article will review the arguments for and against the routine use of antibacterial prophylaxis in HSCT recipients. PMID- 11919725 TI - A phase I/II double-blind, placebo-controlled study of recombinant human interleukin-11 for mucositis and acute GVHD prevention in allogeneic stem cell transplantation. AB - Interleukin-11 (IL-11) decreases cytokine release and increases survival in murine BMT models. In these systems, it reduces gut permeability, partially polarizes T cells to a Th2 phenotype, down-regulates IL-12, prevents mucositis, and accelerates recovery of oral and bowel mucosa. We conducted a randomized double-blind pilot study of rhIL-11 administered with cyclosporine/MTX prophylaxis after cytoxan/TBI conditioning and allogeneic stem cell transplantation for hematologic malignancies. Patients received rhIL-11, 50 microg/kg subcutaneously daily or placebo in a 3:1 ratio. Treatment was administered prior to the start of conditioning and continued up to 21 days. The study was designed to assess safety with stopping rules for cardiac arrhythmias and mortality. Although projected to accrue 20 patients, only 13 patients (10 IL 11, three placebo) were enrolled because the early stopping rule for mortality was triggered. Of 10 evaluable patients who received IL-11, four died by day 40 and one died on day 85. Deaths were attributable to transplant-related toxicity. One of three placebo recipients died of suicide, the other two are alive. Patients receiving IL-11 had severe fluid retention and early mortality, making it impossible to determine whether IL-11 given in this schedule can reduce the rate of GVHD. Grade B-D acute GVHD occurred in two of eight evaluable patients on IL-11 and one of three patients on placebo. The primary adverse events of the study were severe fluid retention resistant to diuresis (average weight gain 9 +/ 4%) and multiorgan failure in five of 10 evaluable patients. The use of IL-11 as GVHD prophylaxis in allogeneic transplantation cannot be recommended as administered in this trial. PMID- 11919726 TI - Quantitative real-time RT-PCR detects elevated Wilms tumor gene (WT1) expression in autologous blood stem cell preparations (PBSCs) from acute myeloid leukemia (AML) patients indicating contamination with leukemic blasts. AB - High-dose chemotherapy with subsequent autologous stem cell transplantation is believed to be of therapeutic benefit in patients with acute myeloid leukemia (AML), especially when no allogeneic bone marrow donor is available. One of the main risks is contamination of the stem cell preparations with leukemic blasts, which may account for a higher relapse rate compared to allogeneic bone marrow transplantation. Since overexpression of WT1 is common in leukemic blasts, we investigated, whether PBSCs from AML patients express WT1 at a higher level as compared to patients with solid cancers. PBSCs of seven patients with AML and of five patients with solid cancers were investigated for WT1 expression. Total WT1 copy count was determined in a standardized quantitative real time RT-PCR. WT1 expression was found in all AML PBSCs with an average copy number of 49.99 +/- 61.09. In solid cancers WT1 expression was statistically significantly lower with a copy number of 3.51 +/- 1.92. In AML patients with sustained complete remission we found a nearly significantly lower WT1 expression than in patients who relapsed within the first year after stem cell transplantation. Our data show a higher WT1 expression in PBSCs of AML patients compared to patients with solid cancers. This finding might indicate a contamination with leukemic blasts. Quantification of WT1 in PBSCs might therefore be useful to estimate the risk of relapse after autologous stem cell transplantation in AML patients. PMID- 11919727 TI - Serial evaluation of the oncological pediatric risk of mortality (O-PRISM) score following allogeneic bone marrow transplantation in children. AB - The O-PRISM score was introduced for risk assessment in children transferred to intensive care following BMT. The aim of this study is to determine the prognostic value of a serial evaluation of the O-PRISM score. Ninety-three children, 58 allogeneic-related and 35 unrelated BMT, were evaluated. At weekly intervals, the O-PRISM was calculated based on the standard PRISM score and the three additional variables CRP, GVHD and hemorrhage. Overall survival was 0.51 +/ 0.05 (48/93 patients). Seventeen children died of recurrent disease and 28 of BMT-related complications. High O-PRISM scores significantly correlated with adverse outcome. The relative risks of DOC of patients with scores > or =10 compared to patients with lower scores were: day 0: 3.9 (95% confidence-interval: 1.1-13.7, P = 0.02), day 7: 2.0 (0.7-6.2, P = 0.20), day 14: 5.2 (1.9-14.0, P = 0.001), day 21: 5.6 (1.9-16.5, P = 0.001), day 28: 11.5 (3.8-100.9, P < 0.001), day 35: 7.3 (1.9-27.7, P = 0.001). As early as day 0, children with scores > or =10 points showed a higher cumulative incidence of DOC than patients with lower scores (0.69 +/- 0.15 vs 0.27 +/- 0.05, P = 0.02). The O-PRISM score represents a useful clinical parameter for serial risk assessment following BMT. As it indicates fatal events early, it may be helpful for parent information and even more for the early establishment of intensified supportive treatment. The O-PRISM score may therefore be a valuable parameter for the evaluation of different strategies for BMT and supportive treatment. PMID- 11919728 TI - Association between pretransplant Thymoglobulin and reduced non-relapse mortality rate after marrow transplantation from unrelated donors. AB - A matched cohort study was designed to test the efficacy of polyclonal rabbit antiserum specific for human T cells (Thymoglobulin), administered in vivo on days 1-5 (2 mg/kg/day) before T cell-replete unrelated donor marrow transplantation. Thymoglobulin was given to 52 leukemic patients at Huddinge Hospital. Control patients matched for diagnosis, disease stage, age and treated with a similar regimen, apart from the omission of Thymoglobulin, were selected in Seattle during the same period (n = 104). All received conditioning with cyclophosphamide and TBI. In the study group all patients received 10 Gy single dose TBI, while the controls were given 12-14.4 Gy fractionated TBI. GVHD prophylaxis was cyclosporine and methotrexate. Patients were treated for grade I acute GVHD in the study group, and for grade II GVHD in the control group. Multivariable analyses were adjusted for patient and donor age and CMV serology, HLA matching, donor gender and marrow cell dose. Non-relapse mortality was lower in the study patients (hazard ratio = 0.30, 95% CI 0.12-0.75, P value = 0.005). The 5-year cumulative incidence of non-relapse mortality was 19% in the study cohort, and 35% in the control cohort. Overall mortality was also lower in study patients (hazard ratio 0.51, 95% CI 0.27-0.97, P value = 0.03). No significant difference in the risk of relapse was seen (P = 0.63). This suggests that Thymoglobulin during conditioning may reduce non-relapse mortality after unrelated donor marrow transplantation. PMID- 11919729 TI - Glucose-6-phosphate dehydrogenase deficiency and hematopoietic stem cell transplantation. AB - Glucose-6-phosphate dehydrogenase (G6PD) deficiency is an X-linked hemolytic enzymopathy affecting 3% of Southern Chinese males. Among 275 adult allogeneic hematopoietic stem cell transplantations (SCT), five cases (1.8%) each of donors and recipients were G6PD deficient. Among 107 autologous SCT, four patients (3.7%) were G6PD deficient. All subjects were male, except for two female patients with chronic myeloid leukemia (CML). The incidence of G6PD deficiency in female CML patients was significantly higher than the background female incidence (P = 0.004), but comparable with that in the males (P = 0.664). There was no significant hemolysis or delay in red cell engraftment, and all but one patient converted to donor G6PD screening status. One female patient achieved partial correction of her G6PD status and relapsed at 10 months. We suggest that G6PD deficiency should be tested for in all marrow donors and recipients in susceptible populations. From our data, there is a suggestion of increased clinical incidence of G6PD deficiency in female patients with multi-lineage clonal marrow disorders. PMID- 11919731 TI - Veno-occlusive disease of the liver in children with solid tumors undergoing autologous hematopoietic progenitor cell transplantation: a high incidence in patients with neuroblastoma. AB - We retrospectively analyzed the incidence and risk factors for veno-occlusive disease (VOD) in 83 consecutive children with solid tumors, who underwent autologous blood or bone marrow (BM) transplantation at UCSF between 1992 and 2000. Forty-one patients were diagnosed with neuroblastoma and 42 had another solid tumor (Ewing's sarcoma, soft tissue sarcomas, germ cell tumors, etc). Patients with neuroblastoma were more likely than patients with other solid tumors (ST) to be < or =7 years of age, to have a decreased serum albumin level, and to have received abdominal radiation and surgery prior to transplant. Patients with neuroblastoma received a different conditioning regimen and a purged stem cell product. Twenty patients (24%) developed VOD. VOD was self limited in 15 (75%) patients and severe in five (25%) patients. Univariate analysis identified the following risk factors for VOD: diagnosis of neuroblastoma (odds ratio 6.1, P < 0.01), abdominal radiation (odds ratio 4.1, P < 0.01), abdominal surgery (odds ratio 4.1, P < 0.01), and age < or =7 years of age (odds ratio 3.3, P = 0.02). Disease status at transplant, intensity of previous chemotherapy, conditioning regimen, progenitor cell source, ALT, AST, albumin level, renal function prior to transplant, or use of amphotericin, growth factor or heparin during transplant, did not affect the incidence of VOD. On multivariate analysis, only the diagnosis of neuroblastoma remained significant (odds ratio 7.8, P = 0.03). Larger studies of patients with neuroblastoma are necessary in order to confirm our findings and better define the risk factors for VOD development in neuroblastoma patients. PMID- 11919730 TI - Acquired factor VII deficiency in hematopoietic stem cell transplant recipients. AB - Acquired factor VII (FVII) deficiency in the absence of vitamin K deficiency, oral anticoagulant therapy, synthetic liver dysfunction, or DIC is rare, with only a handful of cases thus far reported. In the period from 1990 to 1996 we identified eight patients with acquired FVII deficiency, all of whom presented with prolongation of the prothrombin time (PT) in the first 2 weeks following stem cell transplantation (SCT). The mean plasma FVII clotting activity (FVII:c) was 22% (range 8-35%) with an approximately equivalent reduction in FVII antigen (FVII:Ag) level. Mean plasma levels of fibrinogen and factors II, V, IX, and X were normal. Protein C activity was significantly depressed in only one of the three patients in whom it was measured. Several patients experienced bleeding complications, and hemorrhage directly accounted for death in two cases. Veno occlusive disease of the liver developed in three patients. We conclude that FVII deficiency should be considered in the differential diagnosis of prolonged PT in patients who have recently undergone SCT. The mechanism of this acquired deficiency state remains to be defined. PMID- 11919732 TI - Body weight recovery, eating difficulties and compliance with dietary advice in the first year after stem cell transplantation: a prospective study. AB - Among healthcare professionals there is no consensus about the best policy to increase oral intake and promote recovery in the post-hospital phase after bone marrow or blood stem cell transplantation. In order to evaluate body weight recovery and compliance with dietary advice among these patients, we performed a prospective longitudinal study in the first year post transplant. At five time intervals (days 50, 75, 125, 200 and 350) patients received a nutritional questionnaire with items on nutrition-related symptoms, physical condition, body weight recovery and compliance with dietary advice. From the initial cohort of 135 patients 69 completed the study. Prevalence of eating difficulties was high (66% at day 50). Anorexia, dry mouth, altered taste, nausea and tiredness were the symptoms most strongly associated with eating difficulties. Compliance with dietary advice was poor. Conditioning regimen was found to be a prognostic factor for body weight status at day 350. In more than 50% of the TBI-treated patients body weight was not restored to 95% of the pretreatment value within 1 year after transplant. Future studies should focus on increasing energy and protein intake in the TBI-treated subgroup. PMID- 11919734 TI - Acute health-related quality of life in children undergoing stem cell transplant: II. Medical and demographic determinants. AB - Medical and demographic variables were examined as predictors of acute health related quality of life (HRQL), specifically, somatic distress, mood disturbance and activity levels, during the period of bone marrow transplant (BMT) hospitalization, and the transition phase in the months following hospital discharge. The responses of 153 children undergoing BMT were assessed by both parent report and patient self-report in a prospective longitudinal design. Type of transplant, diagnosis, age, gender, and socio-economic status (SES) were examined as predictor variables of patient outcome. Type of transplant, patient age, and SES emerged as significant determinants of patient response. Children undergoing unrelated donor (MUD) transplants experiencing the highest levels of distress, followed by those undergoing matched-sibling BMT, while those undergoing autologous transplant experienced the lowest levels of distress. Younger patients experienced lower levels of distress and better HRQL than older children and adolescents. Although patients from different SES backgrounds appeared very similar at the time of hospital admission, those from lower SES backgrounds demonstrated greater distress and disturbance in HRQL subsequently, and throughout the first 6 months post BMT. These findings help to target specific subgroups of patients that may be in greater need of preventive interventions or more aggressive supportive care. PMID- 11919733 TI - Acute health-related quality of life in children undergoing stem cell transplant: I. Descriptive outcomes. AB - There has been little empirical documentation of the acute effects of bone marrow or stem cell transplant (BMT) on children. In the present study, the responses of 153 children undergoing BMT were assessed in a prospective, longitudinal design. Children were assessed at the time of admission for transplant, then underwent weekly assessments to week +6, followed by monthly assessment to month +6. Data were obtained both by parent report and patient report (for patients age 5 and up) using the BASES scales. The major findings are: (1) children undergoing BMT enter the hospital with an already heightened level of distress (defined by high levels of somatic symptoms and mood disturbance, and low levels of activity) that increases dramatically following conditioning, reaching a peak approximately 1 week following transplant; (2) this increased distress is transient, declining rapidly back to admission levels by week +4 to week +5, followed by a further decline to presumed basal levels by months 4-6; and (3) the trajectories of distress depicted by both parent and child report are remarkably similar, each providing confirmatory support for the validity of the findings. These findings confirm a number of widely held clinical impressions that had not previously been documented empirically, and point to the need for new interventions or more intensive approaches to supportive care aimed at reducing levels of distress during the acute phase of transplant. PMID- 11919735 TI - Role of different medium and growth factors on placental blood stem cell expansion: an in vitro and in vivo study. AB - Expansion of haemopoietic stem cells from placental blood has been obtained with a combination of flt3 ligand (FL), thrombopoietin (TPO), kit-ligand (KL) with or without interleukin-6 (IL6) in serum-replete medium. For clinical use, cell expansion in the absence of serum is a clear advantage. Therefore, stem cell expansion in serum-free (SF) medium with a combination of three (FL, TPO, KL) or four (FL, TPO, KL, IL6) growth factors was compared with the results obtained using fetal calf serum (FCS) or human serum (HS). Human CD34(+) placental blood cells were cultured in the presence of FL, TPO, KL +/- IL6 with SF medium, HS and FCS for up to 8 weeks. CD34(+), CFC, LTC-IC content was measured at intervals. To determine the in vivo repopulating capacity of expanded cells, CD34(+) expanded cells were transplanted in sublethally irradiated NOD/SCID mice. With the three growth factor combination the CD34(+) cell number increased steadily up to the 8 weeks of culture. CD34(+) cells were expanded 67.5-fold with SF, 11.7 with HS and 49.2 with FCS. However, when CFCs and LTC-ICs were considered, a continuous expansion was observed only with HS and FCS, whereas in SF medium after 6 weeks their number started to decline. The addition of IL-6 did not change the expansion significantly. Cells grown ex vivo for 14 days were transplanted into NOD/SCID mice. The engraftment of human cells in mice was higher for serum replete than for SF expanded cells. Nevertheless, SF cultured cells were also able to engraft both marrow and spleen in all animals. In addition, engrafted human cells still maintained clonogenic ability. With KL, FL, TPO +/- IL6 it is possible to expand haemopoietic progenitor cells in a SF medium. Compared with serum-replete cultures, the absolute number of clonogenic cells and in vivo repopulating cells is lower. Although the degree of expansion remains significant, a clinical trial still needs to be carried out to address the question of whether this expansion might be useful in reducing post-transplant aplasia. PMID- 11919736 TI - Acute myeloid leukaemia of donor cell origin developing 5 years after allogeneic bone marrow transplantation for chronic myeloid leukaemia. AB - We report the case of a male patient with Ph-positive CML who developed AML 5 years after allogeneic BMT. Clinically, the AML seemed to develop on the basis of a myelodysplasia. The myeloid origin of blasts has been proven by immunophenotyping. The variable number of tandem repeats (VNTRs) and short tandem repeat (STR) showed donor-type haemopoiesis. The interphase FISH showed the XX genotype directly in the morphologically identifiable blasts and in the CD34 positive sorted bone marrow cells. This proved the new leukaemia to be of donor origin. The necessity of using multiple techniques and the advantage of combined immunophenotyping and FISH methods in this case is emphasized. PMID- 11919737 TI - Leukaemic transformation of donor cells in a patient receiving a second allogeneic bone marrow transplant for severe aplastic anaemia. AB - Allogeneic blood or bone marrow transplantation is a successful treatment for leukaemia and severe aplastic anaemia (SAA). Graft rejection following transplantation for leukaemia is a rare event but leukaemic relapse may occur at varying rates, depending upon the stage of leukaemia at which the transplant was undertaken and the type of leukaemia. Relapse is generally assumed to occur in residual host cells, which are refractory to, or escape from the myeloablative conditioning therapy. Rare cases have been described, however, in which the leukaemia recurs in cells of donor origin. Lack of a successful outcome of blood or bone marrow transplantation for severe aplastic anaemia (SAA), however, is due to late graft rejection or graft-versus-host disease. Leukaemia in cells of donor origin has rarely been reported in patients following allogeneic bone marrow transplantation for SAA. This report describes leukaemic transformation in donor cells following a second allogeneic BMT for severe aplastic anaemia. PCR of short tandem repeats in bone marrow aspirates and in colonies derived from BFUE and CFU GM indicated the donor origin of leukaemia. Donor leukaemia is a rare event following transplantation for severe aplastic anaemia but may represent the persistence or perturbation of a stromal defect in these patients inducing leukaemic change in donor haemopoietic stem cells. PMID- 11919738 TI - Allogeneic blood stem cell transplantation following non-myeloablative conditioning for hypereosinophilic syndrome. AB - A male patient with hypereosinophilic syndrome (HES) underwent an allogeneic blood stem cell transplantation from a female donor following non-myeloablative conditioning consisting of a 2 Gy dose of TBI and fludarabine 30 mg/m(2) intravenously on three days. Before transplantation 100% of the mitotic bone marrow cells of the patient showed a clonal karyotype abnormality. Three months after the transplantation eosinophilia had resolved and a chromosome analysis revealed normal female karyotype. The present study shows that stem cell transplantation after non-myeloablative conditioning may have curative potential for HES. PMID- 11920116 TI - Inferring sex-biased dispersal from population genetic tools: a review. AB - Sex-biased dispersal, where individuals of one sex stay or return to their natal site (or group) to breed while individuals of the other sex are prone to disperse, is a wide-spread pattern in vertebrate organisms. In general, mammals exhibit male-biased dispersal whereas birds exhibit female-bias. Dispersal estimates are often difficult to obtain from direct field observations. Here we describe different methods for inferring sex-specific dispersal using population genetic tools and discuss the problems they can raise. We distinguish two types of methods: those based on bi-parental markers (eg comparison of male/female relatedness, F(st) and assignment probabilities) and those relying on the comparison between markers with different modes of inheritance (eg mtDNA markers and microsatellites). Finally, we discuss statistical problems that are encountered with these different methods (eg pseudoreplication, problems due to the comparison of distinct markers). While the genetic methods to detect sex biased dispersal are now relatively well developed, their interpretation can prove problematic due to the confounding effects of factors such as the mating system of the species. Moreover, the relative power of these methods is not well known and requires further investigation. PMID- 11920117 TI - Wolbachia infection associated with all-female broods in Hypolimnas bolina (Lepidoptera: Nymphalidae): evidence for horizontal transmission of a butterfly male killer. AB - Inherited bacteria that kill male hosts during embryogenesis infect a wide range of insect species. In order to ascertain if there are patterns to host infection, with particular male killing bacteria specialising on particular taxa, we investigated the male killing trait in the butterfly Hypolimnas bolina. All female broods were first reported in this species in the 1920s. Investigation of this system in the Fiji Islands revealed the causal agent of sex ratio distortion in H. bolina to be a male killing Wolbachia bacterium. This bacterium is identical in wsp and ftsZ sequence to a male killer in the butterfly Acraea encedon in Tanzania, suggesting it has moved between host species, yet retained its phenotype. The prevalence of the Wolbachia was calculated for three different island groups of Fiji, and found to vary significantly across the country. Antibiotics failed to cure either the male killing trait or the Wolbachia infection. The implications of these results are discussed. PMID- 11920118 TI - Molecular variation at the self-incompatibility locus in natural populations of the genera Antirrhinum and Misopates. AB - The self-incompatibility system of flowering plants is a classic example of extreme allelic polymorphism maintained by frequency-dependent selection. We used primers designed from three published Antirrhinum hispanicum S-allele sequences in PCR reactions with genomic DNA of plants sampled from natural populations of Antirrhinum and Misopates species. Not surprisingly, given the polymorphism of S alleles, only a minority of individuals yielded PCR products of the expected size. These yielded 35 genomic sequences, of nine different sequence types of which eight are highly similar to the A. hispanicum S-allele sequences, and one to a very similar unpublished Antirrhinum S-like RNase sequence. The sequence types are well separated from the S-RNase sequences from Solanaceae and Rosaceae, and also from most known "S-like" RNase sequences (which encode proteins not involved in self-incompatibility). An association with incompatibility types has so far been established for only one of the putative S-alleles, but we describe evidence that the other sequences are also S-alleles. Variability in these sequences follows the pattern of conserved and hypervariable regions seen in other S-RNases, but no regions have higher replacement than silent diversity, unlike the results in some other species. PMID- 11920119 TI - Dynamics of double and single Wolbachia infections in Drosophila simulans from New Caledonia. AB - The bacterial symbiont Wolbachia can cause cytoplasmic incompatibility in Drosophila simulans flies: if an infected male mates with an uninfected female, or a female with a different strain of Wolbachia, there can be a dramatic reduction in the number of viable eggs produced. Here we explore the dynamics associated with double and single Wolbachia infections in New Caledonia. Doubly infected females were compatible with all males in the population, explaining the high proportion of doubly infected flies. In this study, males that carry only wHa or wNo infections showed reduced incompatibility when mated to uninfected females, compared with previous reports. These data suggest that either the DNA of these bacterial isolates have diverged from those previously collected, or the genetic background of the host has lead to a reduction in the phenotype of incompatibility. Mitochondrial sequence polymorphism at two sites within the host genome was assayed to investigate population structure related to infection types. There was no correlation between sequence polymorphism and infection type suggesting that double infections are the stable type, with singly infected and uninfected flies arising from stochastic segregation of bacterial strains. Finally, we discuss the nomenclature of Wolbachia strain designation. PMID- 11920121 TI - Developmental stability in Brassica cretica: the effect of crossing distance on fluctuating asymmetry in cotyledon morphology. AB - In the present investigation of Brassica cretica, a wild relative of the cultivated cabbage, B. oleracea, we performed an extensive crossing experiment, involving self-pollinations, random outcrosses within populations and hybridizations between populations or species, to evaluate the relationship between crossing distance and developmental stability, estimated as the absolute difference between the right and left lobe of the cotyledons. The frequency distribution of the right-minus-left scores had a narrower peak than expected for normally-distributed data, but there was no directional asymmetry or antisymmetry. Despite evidence for inbreeding depression in seedling biomass and cotyledon size, the type of cross had negligible influence on cotyledon asymmetry. Separate analyses of between-population hybrids revealed differences among progenies from different pairs of populations and a tendency for the F1 hybrid means to decrease with the geographic distance separating the parent populations, but only for the two size variables. Based on these and other observations, we propose that the degree of cotyledon asymmetry is unrelated to genome-wide characteristics, such as the level of heterozygosity and genomic co adaptation, and that cotyledon asymmetry is unrelated to the level of genetic stress experienced by each individual. Hence, there is no reason to consider measures of asymmetry as more sensitive indicators of genetic health than conventional fitness variables. PMID- 11920120 TI - Vertical transmission of Wolbachia in Tetranychus kanzawai Kishida and Panonychus mori Yokoyama (Acari: Tetranychidae). AB - The vertical transmission of Wolbachia in two species of spider mite was investigated and compared. One species, Tetranychus kanzawai Kishida, was infected with a modification negative strain of Wolbachia while the other species, Panonychus mori Yokoyama, was infected with a modification positive strain. The infection showed perfect maternal transmission in the laboratory population of T. kanzawai in which Wolbachia-infected females produced infected offspring regardless of whether they mated with infected or uninfected males, and uninfected females produced Wolbachia-free progenies without regard to the infection status of their mating partners. In artificial P. mori populations initiated with 50% infected and 50% uninfected female adults, the infection frequencies among progenies increased with each generation, reaching 100% at the sixth generation in the Sendai population and after the sixth generation in the Toyama population. In another experiment, in which an artificial T. kanzawai population was composed of 50% infected and 50% uninfected female adults, the infection frequency in progeny populations increased very slowly, reaching 62.5% at the 15th generation. The difference in infection frequency in the two spider mites may be due to the different strains of Wolbachia. PMID- 11920122 TI - Clonal reproduction and population genetic structure of grape phylloxera, Daktulosphaira vitifoliae, in Australia. AB - The grape phylloxera, Daktulosphaira vitifoliae, is a viticultural pest that in the past has devastated vineyards worldwide, yet little is known about this insect's biology. The genetic structure of Australian populations of grape phylloxera and its mode of reproduction were studied following the development of four polymorphic microsatellite loci. Insects were collected from 28 vineyards, with a total of 361 insects included in the study. The majority of vineyards were infested by functionally parthenogenetic lineages of grape phylloxera that inhabit the root system and there was little support for the traditionally described holocyclic life cycle for this species. Clonal diversity was limited in all of the vineyard regions, with the exception of the Rutherglen region. A multiple founder scenario or occasional sex may contribute to diversity within the Rutherglen region. Leaf galling populations comprised classes distinct from the common genotypic classes identified on the roots, suggesting limited exchange between these groups. Implications for the management of D. vitifoliae are discussed. PMID- 11920123 TI - Seed and pollen flow and cline discordance among genes with different modes of inheritance. AB - The relationships between seed and pollen flow and cline discordance/concordance between cytoplasmic and nuclear genes, with the incorporation of the effects of natural selection, are formulated for one locus with two alleles, under assumptions of random mating, no drift and no mutation. Results show that under certain conditions, the relative roles of seed and pollen flow in shaping cline discordance/ concordance are very similar to their roles in influencing population differentiation for selectively neutral markers with different modes of inheritance. Where the disequilibria between cytoplasmic and nuclear genes are of the order similar to selection coefficient, cline discordance/concordance can be predicted from the relative values of the ratio of pollen to seed flow and the ratio of selection coefficients. Where the disequilibria attained by seed and pollen flow are significant, the integrated cytonuclear data are recommended for cline analysis. In both cases, the relative rates of selection coefficients between cytoplasmic and nuclear genes can be roughly estimated according to their characteristic length. PMID- 11920127 TI - Extensions of models for the estimation of mating systems using n independent loci. AB - Inferences about plant mating systems increasingly use highly informative genetic markers, and investigate finer facets of the mating system. Here, four extensions of models for the estimation mating systems are described. (1) Multiallelic probabilities for the mixed selfing-random mating model are given; these are especially suitable for microsatellites; a generalized Kronecker operator is basis of this formula. (2) Multilocus probabilities for the "correlated-matings model" are given; interestingly, comparisons between single- vs multilocus estimates of correlated-paternity can provide a new measure of population substructure. (3) A measure of biparental inbreeding, the "correlation of selfing among loci", is shown to approximate the fraction of selfing due to uniparental (as opposed to biparental) inbreeding; also joint estimation of 1- 2- and 3-locus selfing rates allow separation, under a simple model, of the frequency vs the magnitude of biparental inbreeding. (4) Method-of-moments estimators for individual outcrossing rates are given. Formulae are given for both gymnosperms and angiosperms, and the computer program "MLTR" implements these methods. PMID- 11920128 TI - Heritability of immune function in the caterpillar Spodoptera littoralis. AB - Phenoloxidase (PO) is believed to be a key mediator of immune function in insects and has been implicated both in non-self recognition and in resistance to a variety of parasites and pathogens, including baculoviruses and parasitoids. Using larvae of the Egyptian cotton leafworm, Spodoptera littoralis, we found that despite its apparent importance, haemolymph PO activity varied markedly between individuals, even amongst insects reared under apparently identical conditions. Sib-analysis methods were used to determine whether individuals varied genetically in their PO activity, and hence in one aspect of immune function. The heritability estimate of haemolymph PO activity was high (h(2) = 0.690 +/- 0.069), and PO activity in the haemolymph was strongly correlated with PO activity in both the cuticle and midgut; the sites of entry for most parasites and pathogens. Haemolymph PO activity was also strongly correlated with the degree to which a synthetic parasite (a small piece of nylon monofilament) was encapsulated and melanized (r = 0.622 +/- 0.142), suggesting that the encapsulation response is also heritable. The mechanism maintaining this genetic variation has yet to be elucidated. PMID- 11920129 TI - The evolutionary history of the two karyotypic groups of the common shrew, Sorex araneus, in Poland. AB - Genetic variability within and among two karyotypic groups and five chromosome races of the common shrew (Sorex araneus) in Poland was assayed by sequencing a 1023 bp part of the cytochrome b gene (mtDNA) from 28 individuals. Thirty-four variable positions defined 21 distinct haplotypes with a maximum sequence divergence of 0.88%. No significant differentiation in the cytochrome b gene between Western and Eastern Karyotypic groups was found. Haplotype diversity estimates within the races and groups sampled were high (h = 0.800-0.928), while nucleotide diversity estimates were low (pi = 0.0034-0.0053). The distribution of pairwise nucleotide differences fits well with expectations of a "sudden expansion" model. High haplotype diversity was accompanied by relatively high expected heterozygosity (H(E)) values in nuclear genes (calculated over 47 enzyme loci: H(E) = 0.031 - 0.049), giving no evidence for a recent bottleneck after the process of post-Pleistocene recolonization of Poland by the shrews. Thus, for S. araneus chromosome races in Poland, the data on the cytochrome b gene variability support the hypothesis assuming the Robertsonian fusions having spread into an ancestral acrocentric distribution. PMID- 11920130 TI - Genetic variation in the vulnerable and endemic Monkey Puzzle tree, detected using RAPDs. AB - Araucaria araucana (Monkey Puzzle), a southern South American tree species of exceptional cultural and economic importance, is of conservation concern owing to extensive historical clearance and current human pressures. Random amplified polymorphic DNA (RAPD) markers were used to characterise genetic heterogeneity within and among 13 populations of this species from throughout its natural range. Extensive genetic variability was detected and partitioned by analysis of molecular variance, with the majority of variation existing within populations (87.2%), but significant differentiation was recorded among populations (12.8%). Estimates of Shannon's genetic diversity and percent polymorphism were relatively high for all populations and provide no evidence for a major reduction in genetic diversity from historical events, such as glaciation. All pairwise genetic distance values derived from analysis of molecular variance (Phi(ST)) were significant when individual pairs of populations were compared. Although populations are geographically divided into Chilean Coastal, Chilean Andes and Argentinean regions, this grouping explained only 1.77% of the total variation. Within Andean groups there was evidence of a trend of genetic distance with increasing latitude, and clustering of populations across the Andes, suggesting postglacial migration routes from multiple refugia. Implications of these results for the conservation and use of the genetic resource of this species are discussed. PMID- 11920131 TI - Genetic diversity within and divergence between rare and geographically widespread taxa of the Acacia acuminata Benth. (Mimosaceae) complex. AB - The Acacia acuminata complex is a group of woody shrubs and small trees comprised of three formally described taxa (A. oldfieldii, A. acuminata ssp. acuminata and ssp. burkittii) and two informal taxa ("small seed" and "narrow phyllode") with contrasting geographical distributions within the south-west of Western Australia. In addition, a series of variant populations of possible hybrid origin exist. Population genetic structure was investigated in 25 populations representing the taxa and variants using 16 polymorphic allozyme loci. All taxa and variants exhibited relatively high levels of genetic variation compared with other woody angiosperms with similar geographic distributions. Levels of genetic diversity in the widely distributed ssp. burkittii and "narrow phyllode" taxon were considerably higher (H(e), 0.311 and 0.319, respectively) than expected for widespread woody shrubs. In contrast, the rare and highly restricted A. oldfieldii exhibited significantly lower levels of genetic diversity (H(e), 0.173) compared with the other taxa in the complex, but higher than other rare woody shrubs. Although morphologically close, associations based on genetic distance showed A. oldfieldii was highly divergent from the other taxa (D = 0.85) while including variant populations confused systematic alliances. The unusual placement of some of these populations and high degree of population differentiation (G(ST), 12.7%) supports the suggestion that these may represent a series of hybridisation events between the various taxa. PMID- 11920132 TI - Microsatellite variation in cyclically parthenogenetic populations of Myzus persicae in south-eastern Australia. AB - We examined the population structure of the introduced aphid, Myzus persicae collected mainly from its primary host, Prunus persica, in south-east Australia. Myzus persicae has been present in Australia since at least 1893. Samples were collected in the spring of 1998 from two mainland and three Tasmanian localities and isofemale lines were established in the laboratory. The reproductive mode (life cycle), karyotype and 17-locus microsatellite genotype of each clone were determined. All populations showed significant population differentiation (F(ST) 0.058-0.202) even over small geographic distances (<50 km). All clones were karyotypically normal except for a subset of clones from one site that was exposed to the carbamate insecticide, Pirimor, the week prior to sampling. Those clones were heterozygous for an autosomal 1,3 translocation frequently associated in M. persicae with insecticide resistance. In contrast to other loci and despite being on different chromosomes, loci myz2(A) and M55(A) showed general and significant linkage disequilibrium. These loci may be affected by epistatic selection. We discuss the observed high clonal diversity, moderate but significant population differentiation, general conformance to Hardy-Weinberg equilibria and low linkage disequilibria with particular focus on the global population biology of M. persicae. PMID- 11920133 TI - Extinction of the Edinburgh lineage of the allopolyploid neospecies, Senecio cambrensis Rosser (Asteraceae). AB - Senecio cambrensis is a new allopolyploid species, which originated independently in North Wales and in Edinburgh, Scotland. Despite extensive searches, the species has not been found growing at any of its previously recorded sites in Edinburgh or at other potential sites in the area since 1993. We have concluded that the Edinburgh lineage of S. cambrensis is now extinct and discuss possible causes of its extinction. The lineage was present in Edinburgh from at least 1974 and therefore survived in the wild for a minimum of 19 years. The species remains well established in parts of North Wales. PMID- 11920134 TI - Host age effect and expression of cytoplasmic incompatibility in field populations of Wolbachia-superinfected Aedes albopictus. AB - The Asian tiger mosquito, Aedes albopictus (Skuse), is a known vector of dengue in South America and Southeast Asia. It is naturally superinfected with two strains of Wolbachia endosymbiont that are able to induce cytoplasmic incompatibility (CI). In this paper, we report the strength of CI expression in crosses involving field-caught males. CI expression was found to be very strong in all crosses between field males and laboratory-reared uninfected or wAlbA infected young females. In addition, crossing experiments with laboratory colonies showed that aged super-infected males could express strong CI when mated with young uninfected or wAlbA infected females. These results provide additional evidence that the CI properties of Wolbachia infecting Aedes albopictus are well suited for applied strategies that seek to utilise Wolbachia for host population modification. PMID- 11920136 TI - Genetic differentiation in the Antarctic coastal krill Euphausia crystallorophias. AB - The population genetics of the Antarctic neritic krill species Euphausia crystallorophias was examined by nucleotide sequence variation in its mitochondrial DNA. A 616 base pair region of the cytochrome c oxidase subunit I (COI) gene was screened for mutations by single-strand conformational polymorphism (SSCP) combined with restriction digestion. E. crystallorophias caught in three different regions of the Antarctic coastline were used--two samples from the Mertz Glacier Polynya and one sample each from the western side of the Antarctic Peninsula and from the Davis Sea. Significant genetic differences between krill samples were identified. However, the extent of these differences did not correlate with the degree of geographic separation between the sampling sites. This suggests that the genetic structuring may be the result of small-scale differentiation rather than differentiation between resident populations in separate parts of the Southern Ocean. The possibility that genetic differences between samples within a region are as important as differences between regions has implications for other studies of krill population genetics. PMID- 11920135 TI - Evolution of the secondary structure of the rRNA internal transcribed spacer 2 (ITS2) in hard ticks (Ixodidae, Arthropoda). AB - ITS2 sequences are used extensively in molecular taxonomy and population genetics of arthropods and other animals yet little is known about the molecular evolution of ITS2. We studied the secondary structure of ITS2 in species from each of the six main lineages of hard ticks (family Ixodidae). The ITS2 of these ticks varied in length from 679 bp in Ixodes scapularis to 1547 bp in Aponomma concolor. Nucleotide content varied also: the ITS2 of ticks from the Prostriata lineage (Ixodes spp.) had 46-49% GC whereas ITS2 sequences of ticks from the Metastriata lineage (all other hard ticks) had 61-62% GC. Despite variation in nucleotide sequence, the secondary structure of the ITS2 of all of these ticks apparently has five domains. Stems 1, 3, 4 and 5 of this secondary structure were obvious in all of the species studied. However, stem 2 was not always obvious despite the fact that it is flanked by highly conserved sequence motifs in the adjacent stems, stems 1 and 3. The ITS2 of hard ticks has apparently evolved mostly by increases and decreases in length of the nucleotide sequences, which caused increases, and decreases in the length of stems of the secondary structure. This is most obvious when stems of the secondary structures of the Prostriata (Ixodes spp.) are compared to those of the Metastriata (all other hard ticks). Increases in the size of the ITS2 may have been caused by replication slippage which generated large repeats, like those seen in Haemaphysalis humerosa and species from the Rhipicepalinae lineage, and the small repeats found in species from the other lineages of ticks. PMID- 11920137 TI - The willow bud galler Euura mucronata Hartig (Hymenoptera: Tenthredinidae): one polyphage or many monophages? AB - The nematine sawfly Euura mucronata Hartig (Hymenoptera: Tenthredinidae) induces galls in the buds of over 30 willow species across the Holarctic region. This extensive host range is surprising, since the other Euura gallers are mostly monophagous; thus, the feeding habit of E. mucronata would represent a switch from monophagy to extreme polyphagy. Previous morphological studies have divided E. mucronata into separate species, but the feeding ranges of these species are unknown, and it is even doubtful whether multiple species really exist. To study whether or not E. mucronata consists of cryptic host-associated sibling species, an allozyme study was conducted using gallers collected from six willow species occurring in northern Fennoscandia. Electrophoretic data from seven variable enzyme loci show that: (1) "E. mucronata" probably comprises at least three species with restricted host ranges, but the species may not be completely reproductively isolated from each other; (2) the pattern of host use is not explained by the phylogeny of willows; (3) the pattern of host use is not concordant with the overall chemical similarity of the hosts; and (4) simple allopatric speciation does not appear to explain the host associations. Consequently, it is possible that reasons such as differences in host phenology, habitat, or morphology, are responsible for the limits in host use in the group. PMID- 11920138 TI - Mating system and population genetic structure of an androdioecious tree, Fraxinus lanuginosa Koidz. (Oleaceae) in northern Japan. AB - Models for the maintenance of androdioecy have suggested that selfing of hermaphrodites decreases the frequency of males in a population (the "male frequency"). To test this hypothesis, we used electrophoretic allozyme methods to study the mating system and population genetics of an androdioecious tree, Fraxinus lanuginosa, which exhibits large variations in male frequency among subpopulations in central Hokkaido (northern Japan). We estimated the outcrossing rates by using seeds assayed at three polymorphic loci, and found that the multilocus outcrossing rate (t(m)) increased with increasing male frequency (q) (t(m) = 0.69 to 0.99, q = 0.11 to 0.59). Fixation indices (F(j)) estimated from these t(m) values ranged from 0.006 to 0.185, and were significantly greater than zero in plots with a low male frequency (q or = 0.47). However, fixation indices at the late life stage (F(a)) were not significantly different from zero at five loci (F(a) = 0.056 to 0.101, q = 0.11 to 0.61), and the F(a) values were lower than the F(j) values in several of the plots where both values were measured. These results indicate that inbreeding depression substantially decreases the proportion of selfed progeny in partially self-fertilising subpopulations. A theoretical model for the maintenance of androdioecy showed expected male frequencies significantly lower than the observed values in plots with high male frequency (q = 0.59 to 0.61), although the differences between the expected and observed values of male frequencies were not significant in plots with intermediate or low male-frequencies (q = 0.11 to 0.47). In this paper, the factors are discussed that affect variations in the male frequency among subpopulations of F. lanuginosa based on these findings. PMID- 11920139 TI - Circadian rhythm and time of mating in Bactrocera cucurbitae (Diptera: Tephritidae) selected for age at reproduction. AB - Chrono-biological traits were changed by selecting for life-history traits via a genetic linkage controlling both time-related behavioural and life-history traits. Behavioural traits were compared between lines selected for young (Y lines) and old (O-lines) age at reproduction in the melon fly, Bactrocera cucurbitae (Coquillett). Adults from O-lines, which survive longer than flies from Y-lines, mated later in the day and had a longer period of circadian rhythm in the locomotor activity than those from Y-lines. Flies from F(1) reciprocal crosses had an intermediate time of mating and periods of circadian rhythm between that of the parents, indicating a genetic basis to these traits. The presence of these behavioural differences across the selection lines indicates that chrono-biological traits exhibit correlated responses to selection on age at reproduction. The correlated responses in the behavioural traits to selection for life-history traits are discussed from two points of view: pleiotropy and inadvertent selection. PMID- 11920140 TI - Temperature and genotypic effects on life history and fluctuating asymmetry in a field strain of Culex pipiens. AB - Fluctuating asymmetry (FA) has been proposed as a tool to measure levels of stress experienced by populations of organisms during development. To be of value as a bio-marker to highlight conditions at particular sites, it is important that variation in FA is due to environmental (eg pollution) variation and not genetic variation among populations and families, in other words heritability for FA should be very close to zero. A full-sib design was set up in which families of Culex pipiens mosquitoes collected from the field were reared at three different developmental temperatures. The effects of temperature and family on developmental rate, egg to adult survival and four wing morphological measures were assessed. There was both a temperature and a family effect on development rate and survival. Temperature affected all four wing traits, but an influence of family was only evident in two of the wing traits. Two separate measures of FA for each of the wing traits were obtained. The mean estimates of FA were mainly around 1% of the value of the character measured. There was evidence of an increase in FA with increase in temperature stress. Heritability was estimated for the wing traits and wing trait FA's using restricted estimation maximum likelihood. The estimates of heritability for the wing traits were small and, individually, did not differ significantly from zero. There was also no evidence of heritable genetic variation for any of the wing trait FA's. The results are discussed in relation to other studies where FA heritabilities have been estimated and in relation to the use of FA as an indicator of environmental stress. PMID- 11920141 TI - Microsatellite genetic variation in small and isolated populations of Magnolia sieboldii ssp. japonica. AB - Magnolia sieboldii ssp. japonica, distributed mainly in western Japan, is restricted to high elevation areas (1000-2000 m above sea level) and usually forms small isolated populations. Four microsatellite loci were assayed for 19 populations from six regions spanning the range of distribution, and the levels and distribution of genetic variation were estimated. All four loci were variable, with a total of 39 alleles, but the overall level of microsatellite genetic variation was low, especially compared with a related species, M. obovata. Genetic structure in M. sieboldii was characterised by low intrapopulational genetic variation (A = 3.74 and H(o) = 0.366 on average) and high genetic differentiation even among regional populations. Highly significant isolation-by-distance (IBD) models at the short distance were detected. Genetic drift and limited gene flow was considered to be important in determining the genetic structure within regions. Total genetic differentiation was remarkably high (F(ST) = 0.488 and R(ST) = 0.538), suggesting genetic barriers among regions. Neighbour-joining dendrograms relating the 19 populations, and further analysis on the IBD models, revealed that a stepwise mutation model was more suited than an infinite allele model to explain the genetic differentiation among regions. It is suggested that mutation at microsatellite loci might be influential in generating the genetic differentiation among regions. These results showed the potential of hypervariable microsatellite loci to evaluate the effects of genetic drift and population isolation within regions, and to detect genetic distinctiveness, in spite of the loss of overall genetic variation in M. sieboldii. PMID- 11920142 TI - Phylogeographical structure revealed by chloroplast DNA variation in Japanese beech (Fagus crenata Blume). AB - Intraspecific genetic variation in three non-coding chloroplast DNA (cpDNA) regions (trnT-L and trnL-F spacers, and trnL intron) of Japanese beech (Fagus crenata Blume) was investigated. This species is a major constituent of the typical cool-temperate deciduous forests in Japan. Twenty-one F. crenata populations from throughout Japan, and four F. japonica populations, a close relative of F. crenata, were examined. Seven haplotypes were distinguishable in F. crenata based on nucleotide substitutions and indels. Pairwise nucleotide diversities among haplotypes ranged from 0.0000 to 0.0042 for F. crenata, including F. japonica. The geographical distribution of cpDNA haplotypes was found to be highly structured in F. crenata. Four haplotypes predominated: haplotypes FC1 and FC4 are prevalent on the Pacific Ocean coast, haplotype FC6 is prevalent on the Japan sea coast from the San-in district to Hokkaido, whilst haplotype FC3 is restricted to northern Kyushu and the western-most part of Honshu. Two haplotypes (FC5 and FC7) are restricted to single populations and one haplotype (FC2) is a derivative of FC1. Each of these haplotypes, except FC2, are thought to be derived from different glacial refugia. Phylogenetic analysis showed that neither F. crenata nor F. japonica was monophyletic for the haplotypes, suggesting either ancestral polymorphism or ancient introgression between the lineages of these two Fagus species. PMID- 11920146 TI - Progress in the genetics of autism. PMID- 11920149 TI - Retrograde signaling at central synapses via endogenous cannabinoids. PMID- 11920150 TI - Estrogen and inflammation: hormone generous action spreads to the brain. PMID- 11920151 TI - Beyond the usual suspects: a cholinergic route for panic attacks. AB - For unknown reasons and through poorly understood mechanisms, people at risk of panic attacks are hypersensitive to suffocative stimuli and experience hyperventilation and anxiety after exposure to heightened concentrations of carbon dioxide. Similarly to the physiological reflex response to hypercapnia in animals and man, the anxious response to carbon dioxide in people with panic disorder is at least partially controlled by the central muscarinic receptors. It is suggested here that some modifications of the cholinergic functions could underlie human individual differences in carbon dioxide sensitivity and proneness to experience panic attacks. The hypothesis is based upon experimental evidence that stressful and potentially harmful stimuli prime relatively long-lasting changes in cholinergic genes expression and cholinergic receptors' regulation. The adaptive sequels of these modifications include protection of the brain from overstimulation, and, at the level of the corticolimbic circuitries, promotion of passive avoidance and learning after stress. The extension of the same modifications to the cholinergic receptors involved in chemoception, however, could lower the threshold for reaction to suffocative stimuli, including carbon dioxide. The exaggerated sensitivity to carbon dioxide observed in humans suffering from panic attacks could then be thought of as an evolutionary cost of the involvement of the cholinergic system in shaping otherwise adaptive responses to stress and threatening stimuli. PMID- 11920152 TI - The substituted benzamides and their clinical potential on dysthymia and on the negative symptoms of schizophrenia. AB - In this paper the historical and scientific background that led to the use of substituted benzamides in two apparently unrelated clinical conditions namely dysthymic disorder and schizophrenia will be reviewed, in order to understand if a common mechanism of action may support this dual therapeutic indication. The dopaminergic antidepressant action of substituted benzamides such as sulpiride, has been proposed, since the late 1970s, by several authors and extensively explored in preclinical experiments by our group. In Italy the first marketing authorization obtained for the new substituted benzamide amisulpride, was with the sole indication of dysthymia and therefore a solid clinical experience exists in the use of substituted benzamides in mild forms of depression, with more than 1 000 000 patients being treated in the last 7 years. The proposed mechanism of action of substituted benzamides implies a selective modulation of the dopaminergic system in the mesocorticolimbic area, important for cognitive processing of internal and external cues, related to survival. The selective antagonism of dopamine D2-D3 receptors has been evoked to explain, in small to moderate doses (ie 50-100 mg day(-1)), the antidepressant effect and, in moderate to medium doses (100-400 mg day(-1)), the reported efficacy on negative symptoms of schizophrenia. Thus, substituted benzamides could represent the first class of atypical antipsychotics successfully employed for both depressive states and schizophrenia. Interestingly, recent evidence in the literature suggests that depressive episodes belonging to the bipolar spectrum are among "alternative indications" of other atypical antipsychotics such as olanzapine and risperidone. PMID- 11920153 TI - Organization of the stress system and its dysregulation in melancholic and atypical depression: high vs low CRH/NE states. AB - Stress precipitates depression and alters its natural history. Major depression and the stress response share similar phenomena, mediators and circuitries. Thus, many of the features of major depression potentially reflect dysregulations of the stress response. The stress response itself consists of alterations in levels of anxiety, a loss of cognitive and affective flexibility, activation of the hypothalamic-pituitary-adrenal (HPA) axis and autonomic nervous system, and inhibition of vegetative processes that are likely to impede survival during a life-threatening situation (eg sleep, sexual activity, and endocrine programs for growth and reproduction). Because depression is a heterogeneous illness, we studied two diagnostic subtypes, melancholic and atypical depression. In melancholia, the stress response seems hyperactive, and patients are anxious, dread the future, lose responsiveness to the environment, have insomnia, lose their appetite, and a diurnal variation with depression at its worst in the morning. They also have an activated CRH system and may have diminished activities of the growth hormone and reproductive axes. Patients with atypical depression present with a syndrome that seems the antithesis of melancholia. They are lethargic, fatigued, hyperphagic, hypersomnic, reactive to the environment, and show diurnal variation of depression that is at its best in the morning. In contrast to melancholia, we have advanced several lines of evidence of a down regulated hypothalamic-pituitary adrenal axis and CRH deficiency in atypical depression, and our data show us that these are of central origin. Given the diversity of effects exerted by CRH and cortisol, the differences in melancholic and atypical depression suggest that studies of depression should examine each subtype separately. In the present paper, we shall first review the mediators and circuitries of the stress system to lay the groundwork for placing in context physiologic and structural alterations in depression that may occur as part of stress system dysfunction. PMID- 11920154 TI - Analysis of microsatellite markers at the UCP2/UCP3 locus on chromosome 11q13 in anorexia nervosa. PMID- 11920155 TI - Transmission disequilibrium mapping at the serotonin transporter gene (SLC6A4) region in autistic disorder. AB - The serotonin transporter gene (SLC6A4, MIM 182138) is a candidate gene in autistic disorder based on neurochemical, neuroendocrine studies and the efficacy of potent serotonin transporter inhibitors in reducing ritualistic behaviors and related aggression. An insertion/deletion polymorphism (5-HTTLPR) in the promoter region and a variable number of tandem repeat polymorphism (VNTR) in the second intron, were previously identified and suggested to modulate transcription. Six previous family-based association studies of SLC6A4 in autistic disorder have been conducted, with four studies showing nominally significant transmission disequilibrium and two studies with no evidence of nominally significant transmission disequilibrium. In the present study, TDT was conducted in 81 new trios. A previous finding of transmission disequilibrium between a haplotype consisting of the 5-HTTLPR and intron 2 VNTR was replicated in this study, but not preferential transmission of 5-HTTLPR as an independent marker. Because of inconsistent transmission of 5-HTTLPR across studies, SLC6A4 and its flanking regions were sequenced in 10 probands, followed by typing of 20 single nucleotide polymorphisms (SNPs) and seven simple sequence repeat (SSR) polymorphisms in 115 autism trios. When individual markers were analyzed by TDT, seven SNP markers and four SSR markers (six SNPs, 5-HTTLPR and the second intron VNTR from promoter 1A through intron 2 of SLC6A4, one SSR from intron 7 of SLC6A4, one SNP from the bleomycin hydrolase gene (BLMH, MIM 602403) and one SSR telomeric to BLMH) showed nominally significant evidence of transmission disequilibrium. Four markers showed stronger evidence of transmission disequilibrium (TDT(max) P = 0.0005) than 5-HTTLPR. PMID- 11920156 TI - Mutation screening and imprinting analysis of four candidate genes for autism in the 7q32 region. AB - Genetic studies indicate that chromosome 7q is likely to contain an autism susceptibility locus (AUTS1). We have followed a positional candidate gene approach to identify the relevant gene and report the analysis of four adjacent genes localised to a 800 kb region in 7q32 that contains an imprinted domain: PEG1/MEST, COPG2, CPA1 and CPA5-a previously uncharacterised member of the carboxypeptidase gene family. Screening these genes for DNA changes and association analysis using intragenic single nucleotide polymorphisms (SNPs) provided no evidence for an etiological role in IMGSAC families. We also searched for imprinting mutations potentially implicated in autism: analysis of both DNA methylation and replication timing indicated a normal imprinting regulation of the PEG1/COPG2 domain in blood lymphocytes of all patients tested. The analysis of these four genes strongly suggests that they do not play a major role in autism aetiology, and delineates our strategy to screen additional candidate genes in the AUTS1 locus. PMID- 11920158 TI - Association between a GABRB3 polymorphism and autism. AB - Autistic disorder (OMIM 209850) is a disease with a significant genetic component of a complex nature.(1) Cytogenetic abnormalities in the Prader-Willi/Angelman syndrome critical region (15q11-13) have been described in several individuals with autism.(1) For this reason, markers across this region have been screened for evidence of linkage and association, and a marker (155CA-2) in the gamma aminobutyric acid type-A receptor beta3 subunit gene (GABRB3) has been associated in one study(2) but not others.(3-5) We completed an association analysis with 155CA-2 using the transmission disequilibrium test (TDT) in a set of 80 autism families (59 multiplex and 21 trios). We also used four additional markers (69CA, 155CA-1, 85CA, and A55CA-1) localized within 150 kb of 155CA-2. The use of multi allelic TDT (MTDT) (P < 0.002), as well as the TDT (P < 0.004), demonstrated an association between autistic disorder and 155CA-2 in these families. Meiotic segregation distortion could be excluded as a possible cause for these results since no disequilibrium was observed in unaffected siblings. These findings support a role for genetic variants within the GABA receptor gene complex in 15q11-13 in autistic disorder. PMID- 11920157 TI - Linkage and association of the glutamate receptor 6 gene with autism. AB - A genome scan was previously performed and pointed to chromosome 6q21 as a candidate region for autism. This region contains the glutamate receptor 6 (GluR6 or GRIK2) gene, a functional candidate for the syndrome. Glutamate is the principal excitatory neurotransmitter in the brain and is directly involved in cognitive functions such as memory and learning. We used two different approaches, the affected sib-pair (ASP) method and the transmission disequilibrium test (TDT), to investigate the linkage and association between GluR6 and autism. The ASP method, conducted with additional markers on the 51 original families and in eight new sibling pairs, showed a significant excess of allele sharing, generating an elevated multipoint maximum LOD score (ASPEX MLS = 3.28). TDT analysis, performed in the ASP families and in an independent data set of 107 parent-offspring trios, indicated a significant maternal transmission disequilibrium (TDTall P = 0.0004). Furthermore, TDT analysis (with only one affected proband per family) showed significant association between GluR6 and autism (TDT association P = 0.008). In contrast to maternal transmission, paternal transmission of GluR6 alleles was as expected in the absence of linkage, suggesting a maternal effect such as imprinting. Mutation screening was performed in 33 affected individuals, revealing several nucleotide polymorphisms (SNPs), including one amino acid change (M867I) in a highly conserved domain of the intracytoplasmic C-terminal region of the protein. This change is found in 8% of the autistic subjects and in 4% of the control population and seems to be more maternally transmitted than expected to autistic males (P = 0.007). Taken together, these data suggest that GluR6 is in linkage disequilibrium with autism. PMID- 11920159 TI - Significant dissociation of brain and plasma kinetics with antipsychotics. AB - Current dosing regimens of psychotropic drugs are based on plasma kinetic considerations, although it is unclear whether plasma levels faithfully reflect brain kinetics of drugs.(1,2) To examine this, we compared the kinetics of plasma levels of two widely used antipsychotics, olanzapine and risperidone, vs the time course of their effects in the brain. We used positron emission tomography (PET) and [(11)C]-labeled ligands to quantify striatal and extra-striatal dopamine-2 (D(2)), and cortical serotonin-2A (5-HT(2A)) receptor occupancy in healthy subjects after a single dose, and in patients chronically treated for psychosis. We found a significant dissociation of brain and plasma kinetics. Mean plasma elimination half-lives of single doses of olanzapine and risperidone were 24.2 and 10.3 h, respectively, whereas it took on average 75.2 h with olanzapine, and 66.6 h with risperidone to decline to 50% of their peak striatal D(2) receptor occupancy. We found similar discrepancies between the time course of plasma levels and extra-striatal D(2) as well as 5-HT(2A) receptor occupancy. Our results question the current reliance on plasma kinetics as the main basis for dosing regimens of antipsychotics. Studies of brain kinetics may provide a sounder basis for determining dosing schedules of psychotropic medications. PMID- 11920160 TI - Dopamine depletion results in increased neostriatal D(2), but not D(1), receptor binding in humans. AB - The effect of endogenous dopamine (DA) on neostriatal DA D(1) and D(2) receptor binding potentials (D(1)RBP and D(2)RBP, respectively) in vivo was evaluated with positron emission tomography (PET) and the radiotracers [(11)C]SCH23390 and [(11)C]raclopride, respectively, by comparing the D(1)RBP and D(2)RBP before and after acute DA depletion. DA depletion was achieved by per-oral administration of 4500 mg alpha-methyl-para-tyrosine (AMPT) given in the 25 h prior to [(11)C]SCH23390 PET and of 5250 mg AMPT given in the 29 h prior to [(11)C]raclopride PET. Six healthy subjects completed the protocol. The AMPT treatment decreased plasma levels of the DA metabolite homovanillic acid by 61 +/ 16% (4500 mg; average +/- standard deviation) and 62 +/- 17% (5250 mg), and levels of the norepinephrine metabolite 3-methoxy-4-hydroxyphenethyleneglycol by 58 +/- 7% (4500 mg) and 66 +/- 5% (5250 mg). This AMPT treatment increased D(2)RBP significantly from 3.18 +/- 0.34 to 3.59 +/- 0.30 but no significant change was observed in D(1)RBP (1.64 +/- 0.24 pre AMPT vs 1.70 +/- 0.17 post AMPT). Thus, while DA depletion "uncovers" D(2)receptors, it does not do so for D(1) receptors. The implications of this finding for measuring endogenous DA and its effects on in vivo receptor binding in humans are discussed. PMID- 11920163 TI - Tcf/Lef transcription factors during T-cell development: unique and overlapping functions. AB - The founding members of the TCF family are T-cell factor-1 (Tcf-1) and lymphoid enhancing factor-1 (Lef-1). In adult mammals, Tcf-1 is uniquely expressed in T lymphocytes, while Lef-1 is expressed in T cells and early B cells. During murine development, however, expression of Tcf-1 and Lef-1 occurs in complex overlapping patterns in many tissues. The unique in vivo function of Tcf-1 and Lef-1 have been explored by gene disruption experiments. Tcf-1-/- knockout mice are severely impaired in the generation of T cells, but are otherwise normal. Lef-1-/- mice lack hair, teeth, mammary glands and trigeminal nuclei and as a consequence die around birth. As deduced from direct analyses and from transplantation experiments, the Lef-1 mutation has no major effects on the immune system. In Tcf 1/Lef-1 double knockout mice, development of T cells is completely abrogated, indicating that Lef-1 can substitute for Tcf-1 in T-cell differentiation. Factors of the TCF/LEF HMG domain family (TCFs) exist throughout the animal kingdom. It has become evident that the TCFs interact with the vertebrate Wnt effector beta catenin to mediate axis formation in Xenopus. Likewise, Armadillo (the Drosophila ortholog of beta-catenin) is genetically upstream of Drosophila TCF in the Wingless pathway. Upon Wingless/Wnt signaling, Armadillo/beta-catenin associate with nuclear TCFs and contribute a trans-activation domain to the resulting bipartite transcription factor. In the absence of Wnt signaling, Tcf factors associate with proteins of the Groucho family of transcriptional repressors to strongly repress target gene transcription. PMID- 11920164 TI - Chronic myeloid leukemia cells resistant to interferon-alpha lack STAT1 expression. AB - INTRODUCTION: Interferon-alpha (IFN) plays a role in the management of different neoplasias, particularly those of hematological origin. The mechanisms of action of IFN are still poorly understood and the individual response is unpredictable. In the present study, the pattern of intracellular gene expression following in vitro and in vivo exposure of chronic myeloid leukemia (CML) cells to IFN was evaluated and correlated with the response to in vivo treatment with IFN. MATERIALS AND METHODS: CML patients in different phases of the disease were studied. The pattern of expression of two IFN-inducible proteins involved in IFN mediated biological activities, the p91 and p84 proteins (STAT1alpha and STAT1beta), components of the IFN-stimulated gene factor 3 (ISGF3) complex and the enzyme 2'-5' oligoadenylate synthetase (2'-5' OASE) were investigated by Western blot in peripheral blood mononuclear cells stimulated or not in vitro by IFN. RESULTS AND CONCLUSIONS: In 6/9 patients evaluated before starting treatment, STAT1 was expressed either constitutively or after in vitro stimulation by IFN. In three cases, STAT1 remained negative even after in vitro activation. The pattern of protein expression correlated with the subsequent hematological response to prolonged in vivo IFN administration: the presence of STAT1 being associated with the clinical response to IFN and the absence and non inducibility of STAT1 with resistance to IFN. This was further substantiated by studies carried out in ten patients analyzed at the time of a documented clinico hematological response or resistance to the in vivo administration of IFN. Finally, in order to establish whether the pattern of response to IFN treatment could be predicted at diagnosis, cells cyropreserved at diagnosis from patients with a documented complete response, confirmed also by cytogenetic negativity, or resistance, were studied. While complete responders proved STAT1 positive, none of the four resistant cases ever expressed STAT1. The expression of 2'-5' OASE did not correlate with the clinical response to IFN. This study documents the pivotal role of STAT1 in the in vitro and in vivo responses of CML cells to IFN. The constitutive or induced presence or absence of STAT1 shows a predictive correlation with the response or resistance to treatment with IFN and could be utilized to identify, at diagnosis, resistant patients who may be spared an expensive and unnecessary prolonged IFN administration. PMID- 11920165 TI - Deregulated expression of promyelocytic leukemia zinc finger protein in B-cell chronic lymphocytic leukemias does not affect cyclin A expression. AB - INTRODUCTION: The promyelocytic leukemia zinc finger (PLZF) gene encodes a transcription factor expressed in myeloid, lymphoid and CD34(+) progenitor cells. Structurally related to BCL-6, which is involved in human lymphoma, PLZF may have a role in proliferation, differentiation and survival of hematopoietic cells, that could be mediated by transcriptional repression of the cyclin A gene. MATERIALS AND METHODS: Quantitative competitive reverse transcription-polymerase chain reaction was used to measure the levels of expression of PLZF and cyclin A in normal leukocyte subsets (including CD19(+) lymphocytes, n=21) and malignant B lymphocytes (including B-chronic lymphocytic leukemias [B-CLL], n=63). Results obtained with this method were confirmed by Western and Northern blot analysis. Transactivation assays were performed using an expression construct for PLZF and two cyclin A promoter luciferase reporters in an Epstein-Barr virus (EBV) transformed B-cell line. Cyclin A expression, cell growth kinetics, and cell cycle were analysed in stable clones of the Burkitt lymphoma (BL) B-cell line DG75 with inducible expression of PLZF, generated using the tetracycline regulated expression system. RESULTS: Expression of PLZF was 100-fold downregulated in 90% B-CLL (56/63) compared to normal B lymphocytes (P<0.001). B CLL patients with the highest levels of PLZF had a poorer survival (P<0.013). In transactivation assays, PLZF inhibited the activity of the cyclin A reporters by 50%, demonstrating that PLZF can repress cyclin A expression in non-malignant B lymphocytes. However, in B-CLL patients, the level of cyclin A expression was found to be within the normal range. Altered PLZF function in B lymphoid malignancies was further corroborated in the PLZF-regulatable DG75 clones, where induction of PLZF expression did not significantly alter the levels of cyclin A expression, the cell growth kinetics, or the cell cycle phase distribution. CONCLUSION: The lower survival of patients with the highest levels of PLZF suggests that this protein may be a marker of progression in B-CLL. The absence of co-ordinated regulation of PLZF and cyclin A genes in B-CLL and in a malignant B-cell line may indicate a loss of cyclin A control by PLZF in B-CLL and other B cell disorders. Deregulation of PLZF could thus play a role in B-cell malignancy. PMID- 11920167 TI - Pregnancy-associated risk for venous thromboembolism and pregnancy outcome in women homozygous for factor V Leiden. AB - INTRODUCTION: To evaluate the pregnancy-associated risk of venous thromboembolism and the risk of stillbirth and miscarriage a multicenter, retrospective and controlled study was conducted in women carrying the homozygous factor V Leiden mutation and in an agematched control group of women from the normal population. PATIENTS AND METHODS: In 64 homozygous (median age 44 years, range 21-75 years) and in 52 control women from five different centers data on venous thromboembolism and pregnancy outcome were obtained. RESULTS: The 64 homozygous women had in total 212 pregnancies, the 52 control women had 118 pregnancies. In homozygous women 65% of pregnancies ended with delivery of a viable infant, 15% with fetal loss (3.3% stillbirth, 12% miscarriage) and 20% by pregnancy termination. In the control women 75% of pregnancies ended with delivery of a viable infant, 12% with fetal loss (1.7% stillbirth, 10% miscarriage) and 13% by pregnancy termination. The differences were statistically not significant. Venous thromboembolism occurred significantly more often in the homozygous women, in 4.2% (9/212) during pregnancy and in 4.7% (10/212) after delivery or pregnancy termination. None of the control women had a thromboembolic episode. CONCLUSION: Our data indicate that women with homozygous factor V Leiden have a high probability for a favorable pregnancy outcome. The increased risk for venous thromboembolism during pregnancy and after delivery would favor heparin prophylaxis during and after pregnancy in women homozygous for factor V Leiden. PMID- 11920166 TI - Are myeloma patients with renal failure candidates for autologous stem cell transplantation? AB - INTRODUCTION: Renal function is one of the most important prognostic factors in multiple myeloma (MM). Patients with renal failure are generally excluded from high dose therapy even though they display a poor prognosis with conventional chemotherapy schemes. The aim of this study was to analyze the outcome of MM patients with renal insufficiency undergoing autologous stem cell transplantation (ASCT), including the evaluation of the quality of PB stem cell collections, kinetics of engraftment, transplant-related mortality, response to high dose chemotherapy and survival. MATERIALS AND METHODS: From a total of 566 valuable patients included in the MM Spanish ASCT registry, three groups of patients were defined: group BA, patients with abnormal renal function at diagnosis but normal at transplant (73 cases); group BB, patients with abnormal function both at diagnosis and at transplant (14 cases); and group AA (control group, 479 cases), patients who constantly had normal renal function. RESULTS AND CONCLUSION: Patients from groups BA and BB presented with a significantly higher number of adverse prognostic factors, reflecting that we were dealing with high tumor MM cases, as compared with patients from group AA. The number of mononuclear cells, CD34+ cells and CFU-GM cells collected in patients with non-reversible renal insufficiency was similar to those harvested in MM patients with normal renal function. Moreover, neutrophil and platelet engraftments were identical in patients with and without renal failure (days +11 and +12, respectively). By contrast, transplant-related mortality (TRM) was significantly higher in group BB patients (29%) than in groups BA (4.1%) and AA (3.3%). In multivariate analysis only three variables showed independent influence on TRM: poor performance status (ECOG 3), hemoglobin <9.5 g/dl and serum creatinine > or =5 mg/dl. The response to high dose therapy was independent of renal function. Interestingly, 43% of patients from group BB showed an improvement in renal function (creatinine < 2 mg/dl) after transplant. The three-year overall survival from transplantation was 56, 49 and 61% for the BB, BA and AA groups, respectively, with a statistically significant difference favoring group AA (P<0.01). PFS did not differ significantly between the three groups of patients. In multivariate analysis the only unfavorable independent prognostic factors for overall survival were poor performance status either at diagnosis or at transplant, high beta(2) microglobulin levels, and no response to transplant. According to these results, ASCT is an attractive alternative for MM patients with renal insufficiency, and it should not constitute a criterion for exclusion from transplant unless patients display poor performance status and very high creatinine levels (>5 mg/dl). PMID- 11920168 TI - Recurrent molecular deletion of the 12p13 region, centromeric to ETV6/TEL, in T cell prolymphocytic leukemia. AB - INTRODUCTION: T-cell prolymphocytic leukemia is a rare form of mature leukemia which occurs in adults and in younger patients suffering ataxia telangiectasia. Among others, complex chromosome aberrations of chromosome 12 have been described in this disease. We searched for deletions of the 12p13 region as the result of these chromosome rearrangements. MATERIAL AND METHODS: Paired leukemic and non leukemic cells were obtained from a series of 21 patients suffering T-cell prolymphocytic leukemia. Loss of heterozygosity was searched for by microsatellite typing using a fluorescent automated laser DNA sequencer to analyze the amplification products. Proteins were analyzed by Western blot. Southern blot analysis of one patient was conducted. RESULTS AND CONCLUSION: Loss of heterozygosity of the 12p13 region, including the ETV6 and CDKN1B genes, was detected in nine of these 21 cases (43%). Western and Southern blot analyses of one case demonstrated a biallelic deletion which did not include ETV6. Taken together, our results defined a minimal region of deletion of less than one Mb flanked by the markers b312C2T7 and D12S320, excluding ETV6 as a candidate gene. Deletion of the 12p13 region is thus a highly recurrent genetic event in T-cell prolymphocytic leukemia. PMID- 11920169 TI - Interleukin-6 stimulates HHV-8 replication in bone marrow cultures and infected cell lines. AB - INTRODUCTION: The significance of HHV-8 DNA detection in bone marrow stromal cells from patients with multiple myeloma is still controversial. Since IL-6 plays a key role in the pathogenesis of myeloma, we studied the effect of this lymphokine on HHV-8 DNA detection. MATERIALS AND METHODS: Amplification of HHV-8 DNA from long-term bone marrow cultures established from normal individuals in the presence or absence of 1 ng/ml IL-6 and from an HHV-8 infected ISI cell line. RESULTS AND CONCLUSIONS: IL-6 increased HHV-8 replication in seven of ten bone marrow cultures as well as in the ISI cell line. Quantitative PCR showed a 3-100 fold increase in HHV-8 DNA copy number/microg DNA. These data suggest that when IL-6 is present in the micro-environment, HHV-8 replicates and may be amplified in the absence of systemic infection in patients without cellular immune deficiency. PMID- 11920170 TI - The World Health Organization classification of neoplasms of the hematopoietic and lymphoid tissues: report of the Clinical Advisory Committee meeting--Airlie House, Virginia, November, 1997. AB - INTRODUCTION: Since 1995, the European Association of Pathologists and the Society for Hematopathology have been developing a new World Health Organization (WHO) classification of hematologic malignancies. The classification includes lymphoid, myeloid, histiocytic, and mast cell neoplasms. MATERIALS AND METHODS: The WHO project involves ten committees of pathologists, who have developed lists and definitions of disease entities. A Clinical Advisory Committee (CAC) of international hematologists and oncologists was formed to ensure that the classification will be useful to clinicians. A meeting was held in November 1997 to discuss clinical issues related to the classification. RESULTS: WHO has adopted the 'Revised European-American Classification of Lymphoid Neoplasms' (REAL), published in 1994 by the International Lymphoma Study Group (ILSG), as the classification of lymphoid neoplasms. This approach to classification is based on the principle that a classification is a list of 'real' disease entities, which are defined by a combination of morphology, immunophenotype, genetic features, and clinical features. The relative importance of each of these features varies among diseases, and there is no one 'gold standard'. The WHO classification has applied the principles of the REAL classification to myeloid and histiocytic neoplasms. The classification of myeloid neoplasms recognizes distinct entities defined by a combination of morphology and cytogenetic abnormalities. The CAC meeting, which was organized around a series of clinical questions, was able to reach a consensus on most of the questions posed. The questions and the consensus are discussed in detail below. Among other things, the CAC concluded that clinical groupings of lymphoid neoplasms were neither necessary nor desirable. Patient treatment is determined by the specific type of lymphoma, with the addition of grade within the tumor type, if applicable, and clinical prognostic factors such as the international prognostic index (IPI). CONCLUSION: The experience of developing the WHO classification has produced a new and exciting degree of cooperation and communication between oncologists and pathologists from around the world, which should facilitate progress in the understanding and treatment of hematologic malignancies. PMID- 11920172 TI - Do we treat a disease or a patient? A subtle difference with dramatic consequences. PMID- 11920171 TI - Ethical and legal aspects of placental/cord blood banking and transplant. PMID- 11920173 TI - Choice of replacement therapy for hemophilia: recombinant products only? AB - In the early 1980s it was recognized that coagulation factor concentrates prepared from plasma pooled from thousands of donors transmitted the hepatitis and the human immunodeficiency viruses. The last 15 years have witnessed the development of methods that, applied during concentrate manufacturing, inactivate viruses escaping the screening procedures. The adoption of these measures has dramatically reduced the risk of transmission of bloodborne infections by plasma derived coagulation factor concentrates. The production of recombinant factor VIII and IX and their availability for patient treatment epitomize progress in hemophilia care through DNA technology. No transmission of infectious agents has been documented and the small amount of residual animal proteins, particularly in the early products, has not been associated with any clinically significant adverse reaction. There is little doubt that recombinant factor VIII and IX are the products of choice for the treatment of patients with hemophilia, particularly the second-generation products containing no plasma-derived human protein and fewer and fewer animal proteins. However, availability and cost still oblige most physicians to ration recombinant products and prioritize certain patient groups (previously untreated, previously uninfected). PMID- 11920174 TI - Contact-mediated inhibition of human haematopoietic progenitor cell proliferation may be conferred by stem cell antigen, CD34. AB - INTRODUCTION: The function of CD34, a transmembrane sialomucin expressed by human haematopoietic progenitor cells, is poorly understood. Its structure suggests it may act as a cell adhesion and signalling molecule. MATERIALS AND METHODS: KGIa cells and primary CD34-positive marrow cells were tested for their ability to aggregate in the presence of the anti-CD34 antibody QBEND10; CFU-GM colonies were grown using standard methods and tested for their content of colony-forming cells by replating; 'haematons' were isolated from marrow by filtration; the phosphorylation of CD34 was investigated by immunoprecipitation and Western blotting DISCUSSION: CD34-positive cells in human bone marrow, like KG1a cells, aggregate when incubated with QBEND10. Staining aggregates with anti-CD34-FITC revealed that aggregation involved co-localisation of CD34 at intercellular binding sites. We examined myeloid colonies (CFU-GM) grown from normal human bone marrow cells, and multicellular aggregates ('haematons') separated from freshly aspirated marrow by filtration, and found CD34-positive cells bound together with co-localisation of the CD34 at the binding sites. This finding shows that CD34 positive cell-cell adhesion occurs physiologically in vitro and in vivo. QBEND10 induced aggregation of KG1a and CD34-positive cells was enhanced by staurosporine (a protein kinase C inhibitor) and inhibited by genistein (a protein tyrosine kinase inhibitor). Moreover, aggregated cells had increased phosphorylation of tyrosine on CD34 and translocation of protein kinase C (PKC) to the cytoplasm, compared with non-aggregated cells. We used the ability of primary colonies to produce secondary colonies on replating as a functional parameter and found that the replating ability of the colonies was increased by treatment with genistein (P=0.003). In addition, the ability of individual samples of primary CD34 positive cells to undergo QBEND10-induced aggregation and the ability of CD34 positive cell-derived colonies to produce secondary clones on replating were inversely related (r=0.86). CONCLUSION: Our results suggest that homotypic aggregation of haematopoietic progenitor cells may be an important mechanism for preventing inappropriate proliferation in vivo. Thus, regulation of expression of the CD34 molecule may play an important role in maintaining the normal level of haematopoietic activity by contact-mediated inhibition of progenitor cell proliferation. PMID- 11920175 TI - The impact of autologous stem cell transplantation on the prognosis of mantle cell lymphoma: a joint analysis of two prospective studies with 46 patients. AB - INTRODUCTION: The purpose of this analysis was to investigate if early sequential high-dose therapy with autologous stem cell transplantation (ASCT) can improve the poor prognosis of patients with disseminated mantle cell lymphoma (MCL). PATIENTS AND METHODS: A joint analysis of two parallel single center studies was performed. Both were characterized by a sequential high-dose therapy consisting of an intensive chemotherapy ('HAM' or 'Dexa-BEAM') for mobilization of peripheral blood stem cells and induction of minimal disease followed by a total body irradiation-containing myeloablative regimen and ASCT. Forty-six patients with reference panel-confirmed stage III/IV MCL were included. Thirty-four patients were accrued to the protocol immediately after diagnosis ('upfront ASCT' group). These 34 patients received a standard first-line regimen prior to mobilization. The remaining 12 patients were put on the protocol later during the course of their disease ('delayed ASCT' group). RESULTS: All patients were in remission after mobilization chemotherapy and proceeded to ASCT; there were no exclusions due to poor response, poor mobilization, or patient refusal. With a follow-up of 24 (2-73) months post transplant, the event-free and overall survival probabilities at 2 years were 77 and 100% for the upfront ASCT group compared to 30% (P=0.0007) and 54% (P=0.0016) for the delayed ASCT group. Event free and overall survival tended to be longer in the upfront ASCT group than in the delayed ASCT group also if calculated from initial diagnosis (76 and 93% vs 42 and 63%, respectively, at 4 years after diagnosis; median follow-up 35 months), although this was not statistically significant. Besides timing of ASCT, only spleen size was identified as an independent predictor of survival by univariate and multivariate analysis. CONCLUSION: ASCT is not curative but may improve the prognosis of patients with MCL if performed as part of an intensive first-line treatment strategy. In contrast, the benefits of this approach for salvaging individuals with relapsed disease appear to be limited. PMID- 11920176 TI - Detailed characterization of a complex karyotype in a patient with primary plasma cell leukaemia using multicolour spectral karyotyping and micro-FISH. AB - INTRODUCTION: Plasma cell leukaemia is a rare disorder that usually carries an aggressive course with a rapidly fatal outcome. A variety of chromosomal abnormalities have been reported in plasma cell leukaemia but the clinical significance of an abnormal karyotype is still unclear. MATERIALS AND METHODS: We have applied the molecular cytogenetic techniques multicolour spectral karyotyping and microdissection in combination with fluorescence in situ hybridization on metaphases from a patient with primary plasma cell leukaemia and a fatal outcome. RESULTS AND CONCLUSION: The chromosome analysis showed severe hypodiploidy and 12 marker chromosomes. Identification of the structural rearrangements was not possible using routine cytogenetic methods. Utilizing the methods above, all marker chromosomes could be identified in detail and the karyotype was shown to be very complex. Forty-three breakpoints were found, and 25 could be identified at the band level, among others 14q32 where the immunoglobulin heavy chain locus is situated. Thus, these techniques provide the opportunity to resolve very complex chromosomal changes in a way that has not been previously possible and will consequently be of great importance in the search for hot spots that may harbour new cancer genes. PMID- 11920177 TI - V-region-specific alterations of serum IgM production in multiple myeloma of IgG class. AB - INTRODUCTION: Multiple myeloma is accompanied by decreased polyclonal serum immunoglobulin concentrations. This suppression might be due to non-specific effects on the polyclonal lymphocyte populations as previously suggested, or it could include specific variable region-dependent mechanisms. Thus, differentiation, survival and activation to Ig secretion of B-lineage cells are dependent on the expression and signalling through the variable Ig receptor. The present study addresses the question whether such variable region-specific alterations were present in the peripheral repertoire of IgM antibodies in patients with IgG-secreting MM. MATERIALS AND METHODS: IgM reactivity repertoires towards a large panel of antigens in extracts of homologous tissues (liver, brain, stomach and heart muscle) and bacteria (Bacillus macquarensis) were analysed in sera from 22 patients diagnosed with IgG1 MM. Healthy, matched volunteers served as control donors. A modified Western assay was used, and values obtained from area integration by image analyses were submitted to multiparametric statistics. RESULTS AND CONCLUSION: The results confirm previous observations on the depression of serum IgM concentrations in multiple myeloma, and demonstrate concentration-independent, patient- and V-region-specific alterations in the polyclonal reactivity repertoires. Since the scoring of IgM reactivities by this technique is independent of IgG and because the deviations of IgM reactivity are not coincident with reactivities of (myeloma) IgG in the individual sera, the results indicate that the immunological syndrome of MM includes significant V-region-specific alterations in the polyclonal repertoires of IgM antibodies. PMID- 11920178 TI - Surveillance of Aspergillus galactomannan antigenemia for invasive aspergillosis by enzyme-linked immunosorbent assay in neutropenic patients treated for hematological malignancies. AB - INTRODUCTION: The incidence of invasive fungal infections is increasing in patients with hematological malignancies. Invasive aspergillosis is one of the most frequently encountered infections with a high mortality rate. New diagnostic tests for invasive aspergillosis such as the detection of Aspergillus galactomannan antigen by a sandwich enzyme-linked immunosorbent assay (ELISA) have recently been described. The objective of this study was to evaluate this assay as a potential surrogate for invasive procedures used to diagnose IA. MATERIALS AND METHODS: We analyzed the performance of a commercially available ELISA test which we routinely use for the surveillance of galactomannan antigenemia in patients with hematological malignancies experiencing chemotherapy induced prolonged neutropenia (ANC < 500/mm(3) for more than 7 days). Serum samples were collected on a weekly basis. Test positivity was defined in accordance with the manufacturer's recommendations. RESULTS: Over the 2 year study period, we analyzed 507 samples obtained during 193 neutropenic episodes from 135 patients. Ten, six and two patients were considered to have proven, probable or possible invasive aspergillosis, respectively, based on clinical, radiological or microbiological data. Forty-four positive (Index>1.5) and 26 'undetermined' (1.5 > Index > 1.0) test results were observed in 17 and ten patients respectively. All invasive aspergillosis cases had at least a positive or an undetermined test result. Only one positive and one undetermined result were found in two patients before the onset of clinical or radiological signs suggesting invasive aspergillosis. Sensitivity was 69% and specificity 96% if only positive results are considered; when 'undetermined' test results were combined with positive results, sensitivity attained 100% and specificity 92% suggesting that the cutoff value for positivity can be lowered from 1.5 to 1.0. CONCLUSIONS: Although the ELISA test did not appear to play a role in the early diagnosis of invasive aspergillosis and in the anticipation of antifungal therapy in our experience, it clarifies the diagnosis of infection in probable or possible invasive aspergillosis especially when the cutoff value is lowered and is useful for monitoring patients receiving specific therapy. PMID- 11920179 TI - FISH analysis with a YAC probe improves detection of LAZ3/BCL6 rearrangement in non-Hodgkin's lymphoma. AB - INTRODUCTION: Chromosomal translocations involving the chromosome 3q27 region are common in B-cell non-Hodgkin's lymphoma (NHL), mainly diffuse large cell lymphoma (DLCL) and less often in follicular lymphoma. Most of these rearrangements involve the same major translocation cluster (MTC) on the 3q27 region, disrupting the LAZ3/BCL6 gene. Some of those translocations are difficult to detect by cytogenetic analysis and/or Southern-blot analysis. In the present report we used a FISH assay to improve the detection of LAZ3/BCL6 rearrangements. METHODS: We isolated a YAC clone (803g3), containing the BCL6 gene, in order to analyze by FISH 19 cases of B-cell non-Hodgkin's lymphoma with cytogenetically detectable 3q27 rearrangement, including reciprocal translocation in 11 cases, deletion in two cases, and addition of undefined chromosomal material on 3q27 in six cases. RESULTS: In the 11 cases with reciprocal translocation, FISH results confirmed cytogenetic data and showed disruption of the LAZ3 region: four t(3;4)(q27;p13), two t(3;11)(q27;q23.1), four t(3;14)(q27;q32) and one t(2;3)(p12;q27). In two of the cases, reciprocal t(3;14) was associated with other cytogenetically detectable abnormalities of 3q27, but FISH showed that they did not affect the LAZ3 gene region. FISH demonstrated a reciprocal translocation with LAZ3 gene rearrangement in two of the six patients with add 3q27: one t(3;11) and one t(3;14). In the two patients with del(3q27), one had two 3q27 FISH signals and one had only one 3q27 FISH signal, but no LAZ3 gene rearrangement was observed. CONCLUSION: We have identified a YAC containing the LAZ3/BCL6 gene. This YAC probe could be useful in clinical practice to demonstrate LAZ3 rearrangements by FISH analysis on tumor samples in NHL. PMID- 11920180 TI - Specific dose-response effects of TGF-beta1 on developmentally distinct hematopoietic stem/progenitor cells from human umbilical cord blood. AB - INTRODUCTION: Transforming Growth Factor-beta1 is known to maintain primitive human hematopoietic stem/progenitor cells in a quiescent state. However, its specific role in the control of distinct progenitor cell types needs to be further elucidated. In this study, we have investigated the dose-response effect of TGF-beta1 on progenitors ranging from primitive high proliferative potential (HPP)-Mix, -GM or -BFU-E to later BFU-E, CFU-G or CFU-M. MATERIALS AND METHODS: A clonal semi-solid assay has been used to analyze the effects of a TGF-beta1 blocking antibody (anti-TGF-beta1) or that of active TGF-beta1 added to the medium at concentrations from 10-3000 pg/ml, on these different hematopoietic stem/progenitor cell types. RESULTS AND CONCLUSION: A preferential growth inhibitory effect on the earlier progenitors was observed when low concentrations of TGF-beta1 (10-300 pg/ml) were used. Concentrations of 10-30 pg/ml TGF-beta1 were sufficient to inhibit 90% of the primitive multipotent HPP-Mix, while 100 300 pg/ml TGF-beta1 were required to inhibit 70% of the bipotent HPP-GM and early HPP-BFU-E. TGF-beta1 did not significantly inhibit or activate the growth of later CFU-G and CFU-M, even when added at concentrations 10-100 fold higher. In contrast, a significant growth-inducing effect of very low TGF-beta1 concentrations (< or =30 pg/ml) on a subset of later BFU-E was observed and cannot be explained by a switch of early into later BFU-E. These results emphasize the polyfunctional role of TGF-beta1 in the regulation of hematopoiesis and the need for low, physiological concentrations of TGF-beta1, when studying both the stem cell compartment and more mature progenitor cell subpopulations. PMID- 11920182 TI - An oath for scientists as is The Hippocratic Oath for medical doctors. PMID- 11920181 TI - Phenotypic and functional analysis of the HLA-class I-specific inhibitory receptors of natural killer cells isolated from peripheral blood of patients undergoing bone marrow transplantation from matched unrelated donors. AB - INTRODUCTION: Limited information is available on the natural killer cell reconstitution after bone marrow transplantation and on the possible role of these cells in graft-versus-host-disease. MATERIALS AND METHODS: Blood samples were collected at different time intervals after transplantation. Lymphocytes were analyzed for informative markers by immunocytofluorimetric analysis. Natural killer cells derived from patients undergoing matched unrelated donor transplant were cloned by limiting dilution in the presence of phytohemoagglutinin and IL2. The natural killer cell clones were analyzed for cytolytic activity. RESULTS: In the nine patients analyzed undergoing transplantation from sibling donors, the majority of peripheral blood lymphocytes during the first 80 d after BMT were represented by T lymphocytes, while in the 15 patients undergoing matched unrelated donor transplant natural killer cells consistently outnumbered T lymphocytes. During the early phases after transplantation, most CD56+CD3- natural killer cells did not express CD16 which was expressed at later intervals. Analysis of the inhibitory receptors specific for HLA-class I molecules showed that CD94/NKG2A, specific for HLA-E, unlike normal donors, was expressed by all natural killer cells including the early appearing CD16-negative ones. Killer inhibitory receptors of the Ig superfamily were expressed late and in low percentages after transplantation and were always coexpressed with CD94/NKG2A. Natural killer-cell clones efficiently lysed the HLA-class I-negative cell lines K562 and 721-221. Natural killer-cell populations or clones isolated from patients with graft-versus-host-disease, failed to lyse donor or recipient derived phytohemoagglutinin-induced lymphoblasts. CONCLUSION: Our analysis shows that (1) recipients of matched unrelated donors transplants exhibit a high proportion of NK cells (2) all NK cells express CD94/NKG2A while the HLA-class I specific inhibitory receptors of Ig superfamily appear at later stages and (3) donor NK cells do not lysed donor or recipient target cells. PMID- 11920183 TI - Recombinant erythropoietin therapy as an alternative to blood transfusions in infants with hereditary spherocytosis. AB - INTRODUCTION: In hereditary spherocytosis, erythropoiesis has been described as 'sluggish' during the first months of life. The lack of appropriate erythropoietic response to compensate for increased red cell destruction necessitates blood transfusions in 70-80% of hereditary spherocytosis-affected infants during their first year of life. After this period, less than 30% require regular transfusion support. This transient requirement for transfusion led us to wonder whether anemic hereditary spherocytosis infants, like anemic premature infants, could benefit from recombinant erythropoietin therapy (rHu-Epo). MATERIAL AND METHODS: In 16 hereditary spherocytosis infants (age range 16-119 days) with severe anemia, a compassionate open preliminary study was performed. rHu-Epo treatment (1000 IU/kg/week) was instituted together with iron supplementation. Hemoglobin values and reticulocyte counts were repeatedly assessed. RESULTS: In 13 out of 16 infants, prompt increases in reticulocyte counts were noted after the first week of treatment with 1000 IU/kg/week of rHu Epo. During treatment with Epo these infants maintained clinically acceptable levels of hemoglobin and did not require blood transfusions. As the infants grew and began to mount an adequate erythropoietic response, the rHu-Epo dose could be tapered and the treatment could be discontinued before the age of nine months. CONCLUSION: Epo treatment in most hereditary spherocytosis infants appears to be effective in the management of anemia and could serve as a valuable alternative to packed RBC transfusions. PMID- 11920184 TI - Non-transferrin-bound iron in myelodysplastic syndromes: a marker of ineffective erythropoiesis? AB - INTRODUCTION: Iron overload is usually observed in patients (even untransfused) with myelodysplastic syndromes (MDS), and contributes towards the generation of low molecular weight iron complexes or non-transferrin-bound iron (NTBI), which in turn favors oxidative DNA damage and consequent apoptosis. MATERIALS AND METHODS: Levels of NTBI and lipid peroxidation were evaluated by means of free serum malondyaldehyde (MDA) in untransfused MDS patients and we tried to correlate them with ineffective erythropoiesis, apoptosis and the pattern of in vitro growth. RESULTS: NTBI levels were found to be significantly higher in low risk than in high-risk MDS patients, as well as in patients with a lower myeloid/erythroid ratio. MDA was found to be uniformly higher in the MDS patients as a whole than in normal controls. The bone marrow progenitor cells in the MDS patients with high NTBI levels showed a higher degree of apoptosis, but this difference was not statistically significant. Patients with a leukemic growth pattern had lower NTBI levels than those with a non-leukemic pattern. CONCLUSION: These data suggest that NTBI is related to the degree of ineffective erythropoiesis and that it contributes towards inducing apoptosis in MDS bone marrow precursors. The presence of leukemic growth is associated with low NTBI levels, probably due to increased iron consumption by blast cells. PMID- 11920185 TI - Polymorphonuclear leukocytes released from the bone marrow by granulocyte colony stimulating factor: intravascular behavior. AB - INTRODUCTION: Granulocyte-colony stimulating factor (G-CSF) treatment stimulates the bone marrow and releases polymorphonuclear leukocytes (PMN) into the circulation. This study was designed to measure the intravascular margination, demargination and survival of PMN released from the marrow by G-CSF. MATERIALS AND METHODS: To trace PMN in the circulation, dividing PMN in the bone marrow of rabbits were labeled with 5'-bromo-2'-deoxyuridine (BrdU) and the effects of a single dose of G-CSF (12.5 microg/kg) on the behavior of these labeled cells in the circulation were measured. RESULTS: The results show that G-CSF induced a granulocytosis that peaked 12 h after treatment. This granulocytosis was associated with stimulation of the bone marrow characterized by shortening of the transit time of PMN through the marrow (97.3+/-2.5 h n=4 control vs 78.9+/-3.6 h n=5 G-CSF) particularly in the post-mitotic pool (P<0.01). Morphometric studies of the lung show a reduced sequestration of BrdU-labeled PMN in lung microvessels in G-CSF-treated animals (P<0.05) and a approximately 14-fold (G-CSF-group) vs a approximately 65-fold (control-group) enrichment of BrdU-labeled PMN in lung tissue if compared to circulating blood. The effect of G-CSF on demargination of PMN was measured by transferring BrdU-labeled PMN from donor animals treated with G-CSF to recipients. G-CSF did not cause demargination of intravascular PMN but delayed the clearance of G-CSF-treated PMN in the circulation. This delayed clearance was associated with inhibition of apoptosis in circulating PMN when measured both by morphology (17.7+/-2.3 vs 7.5+/-1.4%, P<0.01) and flow cytometry (16.2+/-1.1 vs 5+/-1.9%, P<0.01) using a DNA end-labeling method (control vs G CSF group). CONCLUSION: We conclude that PMN released from the bone marrow by G CSF sequestered less in the lung microvessels and have a prolonged intravascular life span. PMID- 11920186 TI - Loss of alpha5beta1-mediated adhesion of monocytic cells to fibronectin by interferons beta and gamma is associated with changes in actin and paxillin cytoskeleton. AB - INTRODUCTION: Modulation of the adhesive responses of monocytic cells may reflect their motility within the bone marrow and at sites of inflammation. Monocyte alpha5beta1 integrins mediate fibronectin-dependent adhesion. We previously showed that type II IFN-gamma reduces adhesiveness to fibronectin (Fn) whereas TGF-beta1 enhances cell attachment. Here, we investigate the role of type I IFNs (alpha, beta) on the adhesive capacity of monocytic cells. MATERIALS AND METHODS: The influence of IFNs on the human U937 cell line adhesion to fibronectin-coated surfaces was determined. The expression of integrins and cytoskeleton proteins was analyzed by FACS, Western blotting and/or fluorescence microscopy analyses. RESULTS: IFN-alpha did not affect cell adhesion to fibronectin. In contrast, IFN beta, like IFN-gamma, abrogated U937 adhesion to fibronectin and antagonized TGF beta1-mediated cell attachment to Fn. The impaired binding of IFN-beta- and IFN gamma-treated cells to fibronectin was not due to reduced levels of alpha5beta1 integrins. IFN-beta and IFN-gamma re-organized filamentous actin, and such rearrangement differed from that observed in TGF-beta1-adhesive cells. U937 cells dominantly expressed 44 to 46 kDa paxillin forms and treatment with IFNs enhanced the number of 66 to 70 kDa forms of paxillin. CONCLUSION: Our data show that IFN beta and IFN-gamma induced loss of monocytic adhesion to fibronectin associated with changes in actin and paxillin cytoskeleton, thereby pointing to a possible effect of these cytokines in monocyte trafficking. PMID- 11920187 TI - Cyclin D1 expression in patients with multiple myeloma. AB - INTRODUCTION: Chromosomal abnormalities are detected in 50 to 70% of patients with multiple myeloma (MM). By conventional cytogenetic analysis, a t(11;14)(q13;q32) is observed at a frequency of 3 to 14%. MATERIALS AND METHODS: To demonstrate a cyclin D1 expression in MM patients or MM cell lines, 14 patients with multiple myeloma (MM) and nine human multiple myeloma cell lines (HMCL) were screened by a competitive RT-PCR and/or Northern blot analysis for cyclin D1 expression. Furthermore, we screened 10 MM patients with FISH to demonstrate a relationship between the cyclin D1 expression and the presence of the t(11;14). RESULTS: Five HMCL had a cyclin D1 overexpression: three of them had a t(11;14)(q13;q32) and two had extra copies of chromosome 11. A cyclin D1 expression was found at diagnosis in seven out of 14 untreated MM patients (50%). Out of 14 MM patients, FISH studies were performed in 10 patients. A t(11;14) was detected in three out of 10 patients and extra copies of chromosome 11 were found in two additional patients. CONCLUSION: Cyclin D1 expression is a common event in MM patients (50%) and is associated either with a t(11;14)(q13;q32) or extra copies of chromosome 11. The prognostic role of the cyclin D1 expression and the level of this expression, as compared to other B-cell chronic lymphoproliferative disorders such as mantle cell lymphoma or hairy cell leukemia, remains to be determined in the pathogenesis of multiple myeloma. PMID- 11920189 TI - von Willebrand factor binding to heparin in various types of von Willebrand disease. AB - INTRODUCTION: The purpose was to study von Willebrand factor (vWF) binding to heparin in different types of von Willebrand disease (vWD). MATERIALS AND METHODS: Plasma samples from 92 patients were representative of most vWD subtypes as they included 13 type 1, ten type 2N, 27 type 2A, 23 type 2B, and 19 type 2M patients. We selected assay conditions suitable for the screening of plasma vWF concentrations as low as 15 U/dl vWF:Ag. We determined the range of vWF concentrations in plasma where the percentage of (125)I-MAb/vWF complexes bound to heparin-agarose beads was constant. This range of dilution allowed circumvention of potential competition by other plasma heparin-binding proteins. RESULTS: The multimeric composition of vWF had hardly any influence on the ability of vWF to bind to heparin. Results were expressed as the ratio of heparin binding capacity of patients' plasma to that of normal pool plasma. We found a ratio of 0.99+/-0.004 (mean+/-s.e.m.) for 23 normal individual donors. Furthermore, when comparing the mean values of plasma vWF-heparin binding ratios by ANOVA F-test in the six groups (one normal and five vWD), we found significant differences between them (P<0.0001). Pairwise comparison of multiples by the Scheffe's test indicated that the mean values of ratios in type 2A on the one hand and type 2M on the other, were significantly lower than in normal plasma, type 2N, type 2B and type 1. CONCLUSION: Our data suggest a relationship between the ability of vWF to bind to heparin and to the platelet GPIb receptor, since type 2B and 2N patients have an increased or normal ability to bind to GPIb whereas type 2A and 2M patients have an impaired interaction with that receptor. PMID- 11920188 TI - Thalidomide in patients with advanced multiple myeloma. AB - INTRODUCTION: Recently, a report has suggested the efficacy and safety of thalidomide in refractory multiple myeloma. In an attempt to assess the efficacy and tolerance of thalidomide in advanced multiple myeloma (on behalf of the Intergroupe Franchophone dy Myelome (IFM)), we report the preliminary experience of the IFM with this drug. MATERIALS AND METHODS: Patients with advanced multiple myeloma (n=27) were treated with an oral dose of thalidomide (median 400 mg/day). At the start of treatment, all patients had active disease and 20 patients had received at least one autologous transplantation. RESULTS: Median follow-up was 105 days from the first administration. The serum and/or urine levels of the M component were reduced by at least 75% in four patients including one patient with a >90% reduction, by at least 50% in five patients and by at least 25% in three patients, giving a total response rate of 45% (12 out of 27 patients). Nine patients had stable disease and six patients had progressed disease. Short-term side-effects of thalidomide were generally moderate. CONCLUSION: This study confirms that thalidomide is an effective agent in patients with advanced myeloma. PMID- 11920190 TI - Role of plasminogen activators and urokinase receptor in platelet kinetics. AB - INTRODUCTION: Plasminogen activators (PA) and plasmin are known to affect platelets but little is known of their role in platelet kinetics. We took advantage of genetically deficient mice to explore the role of urokinase (uPA) and tissue type (tPA) PAs, as well as the uPA receptor (uPAR, CD87) in platelet kinetics. MATERIALS AND METHODS: Platelet shape and number were investigated by flow cytometry. Platelet kinetics was investigated by the in vivo biotinylation and FACS analysis. Platelet production was investigated by counting megakaryocytes in bone marrow. RESULTS: Platelets counts were within the same range in wild type (+/+), uPA, tPA and uPAR-deficient mice. Platelet survival was similar in +/+, uPA-/-, tPA-/- but markedly reduced in uPAR-/- mice. The number of megakaryocytes in bone marrow and spleen was increased 2-3-fold in uPAR-/- compared to +/+ mice. TGF-beta mRNA level within the bone marrow was also significantly increased in uPAR-/- mice. Consistent with an increased platelet production, platelets from uPAR-/- mice had a higher RNA content, as seen by Propidium Iodide (PI) labeling and FACS analysis. Since uPAR is detectable in both hemopoietic and non-hemopoietic cells, radiation chimera were prepared. Investigation of platelet kinetics in chimera showed that platelet survival is reduced with a deficit in either bone marrow-derived, or non-hemopoietic, host cells. CONCLUSION: These results demonstrate that uPAR, but not uPA or tPA, is essential for maintaining normal platelet survival. In addition, uPAR-/- mice maintain normal platelet numbers through increased production. PMID- 11920191 TI - Gene expression in CD34(+) cells from normal bone marrow and leukemic origins. AB - INTRODUCTION: To address the molecular regulation of hematopoiesis and the complex mechanism in leukemogenesis, we established the first catalogs of genes expressed in normal bone marrow and leukemia CD34(+) cells. MATERIALS AND METHODS: CD34(+) cell cDNA libraries were constructed using mRNA from adult bone marrow and from a case of acute myeloid leukemia-M5 transformed from myelodysplastic syndrome (MDS-AML). Expressed sequence tags (ESTs) and full length cDNAs were generated by sequencing and were annotated using bioinformatic tools. RESULTS: From a total of 4142 ESTs obtained from normal bone marrow, 3424 meaningful tags were integrated into 1630 clusters, representing 622 known genes, 522 dbEST entries and 486 novel sequences. Out of 5382 ESTs from MDS-AML, 1985 clusters were produced based on the analysis of 4321 useful ESTs, including 711 known genes, 657 known ESTs and 617 novel sequences. Among 251 transcripts found in both bone marrow and MDS-AML EST datasets and those present in only one dataset, 58 showed statistically significant differences in EST copy numbers between the two tissues (P<0.05). Twenty putative full-length cDNAs for novel genes were also cloned from the MDS-AML library. CONCLUSION: The distinct gene expression patterns in MDS-AML-CD34(+) cells as compared to normal control cells may contribute to the development and/or maintenance of the malignant phenotypes of leukemia cells. PMID- 11920193 TI - Phase I trials in cancer treatment. The maximum tolerated dose: a barbarian guideline. PMID- 11920194 TI - STAT3beta does not interfere with granulocyte colony-stimulating factor-induced neutrophilic differentiation. AB - INTRODUCTION: Activation of the signal transducer and activator of transcription protein STAT3 is a crucial step in granulocyte colony-stimulating factor (G-CSF) mediated cell cycle exit and subsequent neutrophilic differentiation of myeloid precursor cells. We have recently demonstrated that this is mediated, at least in part, by upregulation of the cyclin-dependent kinase inhibitor p27(Kip1). The splice variant STAT3beta, that lacks a C-terminal serine residue implicated in the transcriptional activity of STAT3, has been shown to inhibit STAT3-mediated transcription in certain situations. STAT3beta is known to be expressed in hematopoietic cells, but its role in controlling the balance between proliferation and differentiation has not been established. MATERIALS AND METHODS: We ectopically introduced STAT3beta in differentiation-competent 32D cell transfectants expressing human wild type (WT) G-CSF receptors and studied the consequences for G-CSF-mediated responses. RESULTS: Overexpression of STAT3beta did not alter the kinetics of G-CSF-mediated neutrophilic differentiation or p27 induction in 32D/G-CSF-R WT cells. In addition, we found that p27(Kip1) promoter activity was not inhibited by STAT3beta, while inhibition of p27 transactivation by a dominant-negative STAT3 mutant could in fact be alleviated by coexpression of the beta form. CONCLUSION: These findings argue against a role of STAT3beta as a negative regulator of G-CSF-induced expression of p27 and myeloid differentiation. PMID- 11920196 TI - Murine recessive hereditary spherocytosis, sph/sph, is caused by a mutation in the erythroid alpha-spectrin gene. AB - INTRODUCTION: Spectrin, a heterodimer of alpha- and beta-subunits, is the major protein component of the red blood cell membrane skeleton. The mouse mutation, sph, causes an alpha-spectrin-deficient hereditary spherocytosis with the severe phenotype typical of recessive hereditary spherocytosis in humans. The sph mutation maps to the erythroid alpha-spectrin locus, Spna1, on Chromosome 1. MATERIALS AND METHODS: Scanning electron microscopy, osmotic gradient ektacytometry, cDNA cloning, RT-PCR, nucleic acid sequencing, and Northern blot analyses were used to characterize the wild type and sph alleles of the Spna1 locus. RESULTS: Our results confirm the spherocytic nature of sph/sph red blood cells and document a mild spherocytic transition in the +/sph heterozygotes. Sequencing of the full length coding region of the Spna1 wild type allele from the C57BL/6J strain of mice reveals a 2414 residue deduced amino acid sequence that shows the typical 106-amino-acid repeat structure previously described for other members of the spectrin protein family. Sequence analysis of RT-PCR clones from sph/sph alpha-spectrin mRNA identified a single base deletion in repeat 5 that would cause a frame shift and premature termination of the protein. This deletion was confirmed in sph/sph genomic DNA. Northern blot analyses of the distribution of Spna1 mRNA in non-erythroid tissues detects the expression of 8, 2.5 and 2.0 kb transcripts in adult heart. CONCLUSION: These results predict the heart as an additional site where alpha-spectrin mutations may produce a phenotype and raise the possibility that a novel functional class of small alpha spectrin isoforms may exist. PMID- 11920195 TI - Role of early anthracycline dose-intensity according to expression of Philadelphia chromosome/BCR-ABL rearrangements in B-precursor adult acute lymphoblastic leukemia. AB - INTRODUCTION: The use of anthracycline antibiotics in adult acute lymphoblastic leukemia (ALL) has resulted in an improved outcome to remission induction therapy. However,the exact role of these drugs in consolidation therapy is less clear, especially in specific ALL subsets. MATERIALS AND METHODS: A retrospective analysis was conducted on the outcome of 308 patients (median age 35 years, range 13-75) with the most frequent subtype, early-B ALL, treated between 1974 and 1998 on eight consecutive protocols. Anthracycline-related effects were assessed by evaluating the impact of planned anthracycline dose-intensity (A-DI) on long-term outcome. A-DI (in mg/m(2)/week) during the first twelve weeks of consolidation therapy was classified as either "high" (doxorubicin>20, idarubicin>7) or "low". RESULTS: Complete remission was achieved in 78% of cases. With a median follow-up of 6.5 years, on multivariate analysis, disease-free survival (DFS) correlated only with expression of the Philadelphia (Ph) chromosome and/or associated BCR ABL rearrangements (Ph/BCR(+)) (P=0.0001) and planned A-DI (P<0.0001). On this basis, four major prognostic groups with significantly different DFS could be identified: (1) Ph/BCR(-), "high" A-DI (n=102), median 3.5 years and 41% at five years, respectively; (2) Ph/BCR(-), "low" A-DI (n=64), 1.3 years and 16%; (3) Ph/BCR(+), "high" A-DI (n=35), 1.7 years and 20%; (4) Ph/BCR(+), "low" A-DI (n=39), 0.75 years and 0%. When analyzed separately for Ph/BCR(-) (n=166) and Ph/BCR(+) (n=74) patients, the A-DI effect on DFS was preserved in the former (P=0.018) whereas, in Ph/BCR(+) patients, only age <50 years (P=0.004) and blast count <25 x 10(9)/l (P=0.02) correlated with better DFS. However, Ph/BCR(+) patients with the best prognostic profile (age <50 years and blast count <25 x 10(9)/l; n=21) who were treated on "high" A-DI regimens experienced a median DFS of 2.2 years with DFS 21% at five years, compared to 0.67-1 years and 0-10% in other cases (n=53, P<0.01). CONCLUSION: A "high" A-DI may act as a positive treatment-related prognostic factor in early B-lineage ALL. Although mainly restricted to patients with Ph/BCR(-) ALL, A-DI could also influence the outcome in Ph/BCR(+) patients with other favorable prognostic factors. PMID- 11920197 TI - A comparison in normal individuals and sickle cell patients of reduced glutathione precursors and their transport between plasma and red cells. AB - INTRODUCTION: Reduced glutathione is an important antioxidant in red cells whose depletion may contribute to the pathophysiology of sickle cell disease. The current study was designed to examine the availability of reduced glutathione precursors (glutamate, cysteine, glycine and possibly glutamine) together with the activity of the main transport pathways for their uptake (system ASC for cysteine and glycine; system gly for glycine). MATERIALS AND METHODS: Blood samples were obtained from normal (HbAA, HbA cells) and sickle cell disease patients (HbSS, HbS cells); amino acids were measured by HPLC; and transporter activity was measured by radioactive tracer fluxes (using serine and glycine for activity of system ASC; and glycine for that of system gly). RESULTS: Plasma concentrations of cysteine and glycine were increased and concentrations of all amino acids were elevated in HbS cells. The activity of system ASC was increased in HbS cells (both transport capacity and affinity were elevated for serine transport; transport capacity only for glycine). Activity of system gly was also increased (twofold increase in V(max) for glycine flux), though not significantly. Oxygenation also increased the activity of both transporters in normal and HbS cells. CO prevented deoxy-inhibition of glycine transport. Staurosporine (5 microM) inhibited O(2)-stimulated glycine transport through system ASC. It also inhibited the absolute magnitude of transport through system gly, but the O(2)-dependent flux was unaffected. CONCLUSION: Low reduced glutathione levels in HbS cells were not due to decreased substrate availability and O(2) stimulated transport of reduced glutathione precursors in both normal and HbS cells, through a mechanism that is likely to involve Hb and possibly protein phosphorylation. PMID- 11920198 TI - Aplastic anemia and paroxysmal nocturnal hemoglobinuria: a follow-up study of the glycosylphosphatidylinositol-anchored proteins defect. AB - INTRODUCTION: Flow cytometry analysis of peripheral blood cells is a simple and reliable method for establishing the diagnosis of paroxysmal nocturnal hemoglobinuria. The behavior of the clone may vary; increasing or diminishing over time but prospective study of such variations have not been reported so far. MATERIALS AND METHODS: We report herein the results of a prospective follow-up study of 25 patients. Our aims were twofold: first, to evaluate the behavior of the clone (using flow cytometry) over the time; and second, to evaluate if such variations could predict the occurrence of complications or could be used as a tool for monitoring the residual disease after bone marrow transplantation. RESULTS: It was found that flow cytometry can be used to specifically follow the residual disease post allogeneic marrow transplantation in four patients, and that even without transplantation the defective clone can significantly decrease or even disappear (three patients). CONCLUSION: We found that most of the patients did have significant change in the amount of affected cells during more than three years, and that an increased size of the clone poorly predicted the occurrence of complications. PMID- 11920199 TI - Enhancement of erythropoietin-stimulated cell proliferation by Anandamide correlates with increased activation of the mitogen-activated protein kinases ERK1 and ERK2. AB - INTRODUCTION: Anandamide (ANA) is an endogenous ligand for the cannabinoid receptors Cb1 and Cb2 that is able to synergistically stimulate the proliferation of hematopoietic growth factor-dependent blood cells in serum-free culture. To elucidate the mechanisms by which ANA enhances the proliferative responses of hematopoietic cells, we investigated the ANA-mediated effects on proliferation, cell cycling, apoptosis and intracellular signaling of erythropoietin-stimulated 32D/EPO cells. MATERIALS AND METHODS: 32D/EPO cells were cultured serum free to determine the effects of EPO and anandamide on these cells. Proliferation was analyzed by tritiated thymidine incorporation. Apoptosis as well as cell cycle analysis was carried out by flow cytometry. MAPKinase activation was determined by Western blotting, using phospho-specific MAPK antibodies. RESULTS: Simultaneous addition of erythropoietin (EPO) and ANA enhanced DNA synthesis and increased 32D/EPO cell numbers in serum-free culture. Interestingly, ANA did not alter the G1/S transition but it accelerated each of the successive cell cycle phases of EPO-stimulated 32D/EPO cells. Percentages of apoptotic 32D/EPO cells were equally low in cultures supplemented with EPO alone or a combination of EPO and ANA. Both cultures showed enhanced activation of two mitogen-activated protein kinases, namely, extracellular factor responsive kinases 1 and 2 (ERK1/2), as well as the MAPK-target gene protein c-Fos. This fully correlated with the synergistic stimulation of proliferation of 32D/EPO cells by EPO and ANA. ANA had no effect on EPO-induced STAT-5 activation of 32D/EPO cells. Experiments with the Cb2 receptor-specific antagonist SR144528 demonstrated that the synergistic stimulation of proliferation by ANA was partially Cb2 receptor mediated. CONCLUSION: These data suggest that the positive effects of ANA on the erythropoietin-induced proliferation of 32D/EPO cells are mediated by receptor dependent as well as receptor-independent mechanisms, both of which involve activation of the mitogen-activated protein kinases, ERK1/2. PMID- 11920200 TI - Molecular basis of haemoglobinopathies and G6PD deficiency in the Comorian population. AB - INTRODUCTION: The Comoro archipelago is characterised by a high prevalence of red cell genetic disorders such as G6PD deficiency and haemoglobinopathies, being a region endemic for malaria. Over the last 15 years, the city of Marseilles in France has become the main destination for Comorian immigrants. This Comorian community includes patients with sickle cell disease, sickle cell/beta thalassaemia trait, thalassaemias and G6PD deficiency. MATERIALS AND METHODS: Allele frequencies for haemoglobin S, beta-thalassaemia and G6PD deficiency were determined from neonatal and prenatal screenings of the Comorian community. Haemoglobin fractions were detected by isoelectrofocalisation, and the quantitation of HbS, HbA, HbA(2) and HbF was performed by cation exchange high performance liquid chromatography. The molecular study involved 31 alleles carrying the betaS mutation (Cd 6 [A-->T]), six beta-thalassaemic alleles and 17 G6PD-deficient alleles, selected from a group of carriers or affected subjects. RESULTS: Allele frequencies were 3% for haemoglobin S, 1% for beta-thalassaemia trait and 9.5% for G6PD deficiency. Molecular analysis had revealed that the African alleles are predominant, being present in almost all the subjects studied. Mediterranean alleles were found for all the beta-thalassaemia mutations and for three G6PD chromosomes out of 17. CONCLUSION: These data are consistent with the mixed Arab and African origin of the population of the Comoro Islands, and are of clinical interest in prenatal and newborn screening plans. PMID- 11920201 TI - Factor XIII Val34Leu polymorphism in primary intracerebral haemorrhage. AB - INTRODUCTION: Recently, the common Val34Leu polymorphism of the A-chain factor XIII gene, associated with high factor XIII activity, has been identified as a protective genetic factor against occlusive arterial and venous diseases. Moreover, this polymorphism has been suggested to be the first one to increase the risk of cerebral haemorrhage in a small number of Caucasian patients. The aim of our study was to investigate the role of this polymorphism in patients with primary intracerebral haemorrhage from a distinct population. MATERIAL AND METHODS: Patients with non-traumatic primary intracerebral haemorrhage (n=116), age-, race-, sex- and risk factor-matched controls (n=116), and individuals from the general population (n=465) were genotyped for the factor XIII Val34Leu polymorphism by polymerase chain reaction and allele specific restriction assay. The relationships of the Val/Leu genotype with distinct intracerebral haemorrhagic risk factors and with early mortality associated with the haemorrhagic episode were also analysed. RESULTS: No statistical difference in terms of prevalence was detected between patients (P=0.190) and controls (P=0.181). The frequency of the FXIII Leu34 allele was similar in the general population (P=0.191). CONCLUSION: The results suggest that the Leu 34 allele of the A-chain factor XIII gene has a minor role in the development of non-traumatic primary intracerebral haemorrhage. Moreover, the simultaneous presence of the Leu 34 allele with selected risk factors for this disease does not increase the risk of developing this disease. PMID- 11920202 TI - Immunotherapy by non-myeloablative stem cell transplantation: study of the immune reconstitution. Arguments for distinct cell subsets in skin and blood. AB - INTRODUCTION: Non-myeloablative peripheral stem cell transplantation has been shown to induce tumour rejection in patients with acute leukaemia. However, the immunological mechanisms involved and the immune reconstitution achieved have not been investigated. MATERIALS AND METHODS: We describe the cases of two patients for whom we have studied the lymphocyte reconstitution achieved, using both phenotypic and genetic analyses of the T-cell repertoire, after peripheral stem cell transplantation. RESULTS: : In both cases we observed immune reconstitution with T-cell repertoire evolution and presence of activated CD8(+) T cells. In one of the patients an activated clone expressing Vbeta8 represents 46% of the CD8(+) cells. Expansion of this clone occurred in the absence of graft vs host disease symptoms. In the second case a skin lesion typical of graft vs host disease appeared after complete remission had been achieved. The T-cell repertoire in a biopsy of the lesion was distinct from that observed in the blood. CONCLUSION: Our study indicates that peripheral donor cells can effectively reconstitute a grafted patient while inducing an immune response against antigens expressed by the leukaemic/myeloma cells. Our data provide arguments for different populations of T cells associated with graft vs leukaemia/lymphoma and GVH effects. PMID- 11920204 TI - Is there a limit to the normality of blood products? PMID- 11920203 TI - Is escalated BEACOPP a standard therapy for advanced Hodgkin's disease? PMID- 11920205 TI - 14q32 chromosomal translocations: a hallmark of plasma cell dyscrasias? PMID- 11920206 TI - Increased erythropoietin level induced by hydroxyurea treatment of sickle cell patients. AB - INTRODUCTION: Administration of hydroxyurea in sickle cell disease is associated with a dramatic increase of HbF along with a significant clinical improvement and, occasionally, increased total hemoglobin levels. The underlying mechanisms are not yet fully elucidated. MATERIALS AND METHODS: We report the response of three patients with homozygous sickle cell disease and 10 patients with compound HbS/beta-thalassemia (four with beta(o)thal/HbS and six with beta(+)thal/HbS respectively) to hydroxyurea treatment with regards to their serum erythropoietin levels (sEpo). RESULTS: Baseline sEpo levels varied from 33.0 to 284.0 IU/L and showed a significant negative correlation with the respective Hb values (P<0.007). Two to three weeks after initiation of treatment, the sEpo values started to increase and reached levels three to 31 times higher than the baseline two to three weeks later. Thereafter, in most cases the Epo values decreased and remained at intermediate levels throughout the rest of hydroxyurea administration, while in a few cases, they returned to baseline. An inappropriate increase of sEpo following treatment with various cytostatic drugs, independently of anemia induced by cytostatic agents, has already been reported in the literature. The cytostatics included cyclophosphamide, anthracyclines, cytosine arabinoside etc., but not hydroxyurea. The results described here with hydroxyurea are virtually similar, ie, they show a significant sEpo increase five to ten days post therapy with no apparent cause. Pulses of high dose Epo have been reported to promote proliferation of erythroid precursors with HbF synthesizing capacity. CONCLUSION: Our hypothesis is that a similar phenomenon may occur here also, in the sense that peaks of endogenous Epo may promote proliferation of erythroid precursors which maintain the capacity to synthesize HbF. PMID- 11920207 TI - Prognostic relevance of a scoring system based on clinical and biological parameters in early chronic lymphocytic leukemia. AB - INTRODUCTION: Among patients with indolent form of B-cell chronic lymphocytic leukemia, some of them will progress into more advanced stages. To better define this subpopulation of patients, we attempted to define some parameters capable of predicting a pejorative clinical outcome. MATERIALS AND METHODS: Eighty-eight previously untreated patients with B-cell chronic lymphocytic leukemia in Binet stage A were analysed to study the prognostic value of simple serological variables: soluble CD23 (sCD23), beta2 microglobulin (beta2m), lactate dehydrogenase activities and albumin level. Results were compared to other conventional clinical and biological parameters by univariate and multivariate statistical analysis. RESULTS: Our data show that: (1) among those studied, sCD23 >50 u/ml was the only serological significant parameter clearly correlated with disease progression and (2) stage A" patients (hemoglobin level between 100 and 120 g/l and/or lymphocytosis >30.10(9)/l), axillary lymph nodes and hypogammaglobulinemia were found to be other variables associated with a pejorative outcome. These four variables enabled the establishment of a scoring system, capable of predicting disease progression since 66% of the patients with a score < or =2 are going to evolve into advanced stages vs 12% with a score <2. Furthermore, the time to progression is shortened when the score is increasing. CONCLUSION: Our findings show the prognostic relevance of a scoring system including sCD23 level. This score could be taken into account in the treatment strategy of B-cell chronic lymphocytic leukemia. PMID- 11920208 TI - Normal and chronic phase CML hematopoietic cells repopulate NOD/SCID bone marrow with different kinetics and cell lineage representation. AB - INTRODUCTION: Chronic myelogenous leukemia is characterized by a clonal expansion of abnormal hematopoietic cells, which eventually replaces normal hematopoiesis. We wanted to test the hypothesis that the growth kinetics of CML and normal hematopoietic cells are different. MATERIALS AND METHODS: We compared the growth kinetics and the phenotype of engraftment of chronic phase CML and normal human CD34(+) precursor cells in the bone marrow of immune deficient mice. RESULTS: High levels of engraftment of normal precursors occurred early and consisted of myeloid, erythroid, megakaryocytic, and lymphoid elements. This level and pattern of engraftment were maintained at later assessments. The level of CML cell engraftment was initially much lower, but it increased progressively at late time points with no indication of a plateau in growth. Early engraftment of CML cells consisted almost entirely of myeloid and mast cells but soon after only mast cells were detectable. Conversely mast cells were infrequent in mice engrafted with normal progenitors. CONCLUSION: We conclude that in contrast to normal cell engraftment, engraftment of CML cells in NOD/SCID mice is characterized by a slow but progressive myeloid infiltration, which eventually consists almost entirely of mast cells. PMID- 11920209 TI - Functional G-CSF pathways in t(8;21) leukemic cells allow for differentiation induction and degradation of AML1-ETO. AB - INTRODUCTION: Efficacy of differentiating agents requires that their specific cellular targets are still expressed and functional in the leukemic cells. One hypothesis to target sensitive cells is to select leukemic clones which harbor disrupted transcription factors. CBFalpha and CBFbeta are core-binding proteins which have been identified as transcription regulators of hematopoietic genes and shown to be altered in numerous leukemias. In M2 AML, the t(8;21) translocation, CBFalpha (AML1) is altered and produced as the AML1-ETO fusion protein. The fusion protein blocks transcription and differentiation mediated by G-CSF. Interestingly, AML1-ETO leukemic cell lines are sensitive to numerous cytokines in vitro and can be induced to differentiate in the presence of G-CSF and PMA. MATERIALS AND METHODS: As in the APL differentiation model, primary culture provides a useful tool for therapeutic screening of differentiation inducers, we analysed the in vitro sensitivity of 10 fresh M2 AML t(8;21) leukemic samples to G-CSF and the functionality of G-CSF intracellular pathways. In vitro data were compared with in vivo data from four patients treated with rhG-CSF at the dosage of 5 microg/kg/day i.v. for two to three weeks before the initiation of AML induction chemotherapy and immunophenotypic analysis performed weekly to monitor in vivo differentiation. RESULTS: In vitro, an increase in CD34+ cells expressing differentiation antigens (CD11b, CD13 or CD15) was noted along with a decrease of immature CD34+/differentiation antigen negative cells. After two weeks of a daily rhG-CSF administration in vivo, a significant, albeit transient, decrease of blast count was achieved, concomitant with an increase in differentiated leukemic cells suggesting that in vivo differentiation occurs. Fresh t(8;21) leukemic cells possess functional G-CSF signaling pathways as normal activity and kinetics of STAT1 and STAT3 binding was observed. Furthermore, differentiation induction leads to a subsequent degradation of the AML1-ETO oncoprotein. CONCLUSION: The data presented here supports the claim that G-CSF can induce in vitro and in vivo differentiation of M2 AML t(8;21) cells. PMID- 11920210 TI - Acute leukemia following a previous malignancy: do acute lymphoid leukemia and acute myeloid leukemia have common risk factors? AB - INTRODUCTION: Within the framework of the GIMEMA Study Group, the characteristics of acute lymphoid leukemia and acute myeloid leukemia occurring in patients who have suffered a previous malignancy were studied. Assessment was also made of the clinical course, laboratory features and overall outcome of these conditions. MATERIALS AND METHODS: A four-year, multi-center retrospective study was conducted to evaluate the effect of treatment for previous hematological malignancy on the development of secondary leukemia. The study collected in the GIMEMA Archive of Adult Acute Leukemia 3934 new cases of acute leukemia (2964 AML, 901 ALL, 60 acute biphenotypic leukemia). Among these cases, data were evaluated from patients with a personal history of a previous malignancy, and included inquiring into demographic data, history of neoplastic diseases in the 1st degree relatives, type and treatment of the previous malignancy, latency until the development of a secondary acute leukemia diagnosis, laboratory features, treatment and outcome at the onset of secondary acute leukemia. RESULTS: Approximately 200 (5.1%) patients presented a previous malignancy. Twenty-one were affected by ALL and 179 by AML. The proportion of patients with secondary AML was higher than that of patients with secondary ALL (179/2964 vs 21/901, O.R. 2.69-95% C.I. 1.66-4.39, P<0.001). The median latency, from the onset of the previous malignancy to the development of secondary ALL was 27 months and to the development of secondary AML was 52 months (P<0.05). Furthermore, of patients who previously received chemotherapy more developed a second AML (66/127 sAML vs 5/21 sALL; O.R. 3.46-95% C.I. 1.10-11.56, P<0.01). CONCLUSION: In most cases, chemotherapy treatment for a previous malignancy can play a role in the development of secondary AML. In almost all cases of secondary ALL, the role of previous drugs does not appear to be relevant. On the basis of our analysis, performed systematically for the first time on a large adult series of acute leukemia, we conclude that in these patients a biological predisposition to cancer may be suspected. PMID- 11920211 TI - Hematopoietic stem cell transplantation in Europe 1998. AB - INTRODUCTION: Transplantation of hematopoietic stem cells from blood or bone marrow has become accepted therapy for many diseases. Numbers of transplants have increased significantly and stem cell source, donor type and indications have changed during this decade. Information on these changes is essential for interpretation of current data, patient counseling and health care planning. PATIENTS AND METHODS: Since 1990, members of the European Group for Blood and Marrow Transplantation and teams known to perform blood or marrow transplants have been invited annually to report their transplant numbers by indication, donor type and stem cell source. Data from these surveys have been used to present data for 1998, to assess current status and to give numbers of transplants per participating country, coefficients of variation between countries for individual indications and changes in indication, stem cell source and donor type over the past decade. RESULTS: In 1998, a total of 20 892 transplants were performed by 528 teams in 31 European countries. Of these transplants 18 400 were first transplants, 5308 (29%) were allogenic, and 13 092 (71%) were autologous. Of the autologous transplants, 809 (6%) were bone marrow derived, and 12 283 (94%) were from peripheral blood stems cells. Of the allogeneic transplants, 3372 (64%) were bone marrow derived, and 1936 (36%) were peripheral blood stem cell transplants. In 1990, the respective figures were 2137 allogeneic (50%) and 2097 (50%) autologous transplants, all exclusively bone marrow derived. Main indications in 1998 were leukemias with 6015 transplants (33%), 68% thereof allogeneic transplants; lymphomas with 7492 transplants (41%), 94% thereof autologous transplants; solid tumors with 4025 transplants (22%), 99% thereof autologous transplants; non-malignant disorders with 868 transplants (5%), 80% thereof allogeneic transplants. Absolute numbers of transplants per year did increase from 4234 in 1990 to 20 892 in 1998. Increase is higher for autologous, than for allogeneic transplants. There were differences in absolute or relative increase over time for individual indications. Transplant rates per number of inhabitants varied between countries, ranging from 0 to >500 total transplants per 10 million inhabitants with a clear correlation between number of teams and transplants per 10 million inhabitants (r=0.61, P<0.001). The least variation between countries was observed for acute leukemias, chronic myeloid leukemia and severe aplastic anemia in allogeneic transplants, for Hodgkin's disease and non-Hodgkin's lymphoma in autologous transplants. CONCLUSION: These data reflect the current status of blood and marrow transplantation in Europe. They show the continuing increase in utilization, highlight the change from bone marrow to blood as stem cell source and give an objective assessment on presence or absence of trends. PMID- 11920212 TI - The 5TMM series: a useful in vivo mouse model of human multiple myeloma. PMID- 11920213 TI - Prevention of postsplenectomy sepsis: how much do patients know? AB - INTRODUCTION: Asplenia causes a deficiency in immunity with a long-term risk of fulminant infection, associated with significant mortality. Patient compliance requires an understanding of risks of infection and its prevention. The impact of patient education has been little studied. MATERIALS AND METHODS: To ascertain the degree of knowledge held by patients who have undergone splenectomy, a comprehensive survey was designed. This also aimed to determine which group of health professionals was most successful in conveying information to patients. Patients who had undergone total splenectomy were interviewed by telephone, using a standardised list of questions to assess their understanding of the post operation risks. RESULTS: Of 40 consecutive patients, 32.5% had a good knowledge of the risks of asplenia and their prevention, 52.5% had a fair knowledge and 15% a poor knowledge. Haematologists were most successful in initially conveying information to patients, and general practitioners also played a critical role in patient education. In this survey, it appears that surgeons were not effective at educating patients. CONCLUSION: Patient education postsplenectomy is poor. Measures to prevent infection in the asplenic patient are not being adequately implemented. PMID- 11920214 TI - Medical practice is actually dictated by lawyers. PMID- 11920215 TI - Reactive plasmacytoses, a model for studying the biology of human plasma cell progenitors and precursors. PMID- 11920216 TI - Factors which predict unsuccessful mobilisation of peripheral blood progenitor cells following G-CSF alone in patients with non-Hodgkin's lymphoma. AB - INTRODUCTION: High-dose therapy with haematopoietic progenitor cell support has increasingly been utilised for patients with haematological malignancies. Peripheral blood is the stem cell source of choice, however, various mobilisation strategies are used by different centres. PATIENTS AND METHODS: Over a 2-year period, 52 patients with non-Hodgkin's lymphoma (median age 47 years, range 16-64 years) underwent peripheral blood progenitor cell mobilisation using G-CSF alone (16 microg/kg/day). The harvest was considered successful if > or =1 x 10(6) CD34(+) cells/kg were collected by leukapheresis. The histological subtypes of non-Hodgkin's lymphoma comprised: follicular (24 patients), diffuse large B-cell (14 patients), lymphoplasmacytoid (four patients), mantle cell (three patients), lymphoblastic lymphoma (one patient) and small lymphocytic lymphoma/chronic lymphocytic leukaemia (six patients). The median interval from diagnosis of non Hodgkin's lymphoma to mobilisation was 27 months (range 2 months to 17 years). The median number of prior treatment episodes was 2 (range 1-5); 26 patients had received fludarabine alone or in combination. At the time of peripheral blood progenitor cell mobilisation, 20 patients were in 1st remission and 32 were in > or =2nd remission; 30 patients were in partial remission and 22 were in complete remission; the bone marrow was involved in nine patients. RESULTS: Peripheral blood progenitor cell mobilisation/harvest was unsuccessful in 19 out of 52 (37%) patients (mobilisation: 18, harvest: 1). The factors associated with unsuccessful mobilisation or harvest were: prior fludarabine therapy (P=0.002), bone marrow involvement at diagnosis (P=0.002), bone marrow involvement anytime prior to mobilisation (P=0.02), histological diagnosis of follicular, mantle cell, or lymphoplasmacytoid lymphoma, or small lymphocytic lymphoma/chronic lymphocytic leukaemia (P=0.03) and female gender (P=0.04). CONCLUSION: Although peripheral blood progenitor cells can be successfully mobilised and harvested from the majority of patients with non-Hodgkin's lymphoma after treatment with G-CSF alone, the latter is unsuccessful in approximately one-third of patients. These factors should be taken into account when patients are being considered for high dose treatment. PMID- 11920217 TI - Autologous peripheral blood progenitor cell transplantation with <2 x 10(6) CD34(+)/kg: an analysis of variables concerning mobilisation and engraftment. AB - INTRODUCTION: This study analyses the factors affecting mobilisation and engraftment in autologous peripheral blood progenitor cell transplantation according to the number of CD34(+) re-infused. MATERIALS AND METHODS: A total of 190 patients underwent mobilisation with G-CSF alone (n=113) or in combination with chemotherapy (n=77). A total of 116 patients (61%) were autografted with <2 x 10(6) CD34(+) cells/kg and 74 patients were transplanted with >2 x 10(6) CD34(+) cells/kg. Rates of granulocyte and platelet recovery were estimated using the product-limit method of Kaplan-Meier and compared using a log-rank test. The Cox regression model was used for the multivariate analysis of factors influencing engraftment. Differences between cohorts were evaluated by one-way ANOVA or Mann-Whitney tests, and multivariate analysis was performed using a stepwise lineal regression. RESULTS: Neutrophil and platelet engraftment was significantly longer with <2 x 10(6)/CD34(+)/kg (12 vs 10 days, P=0.014 and 16 vs 13 days, P=0.0001 respectively). Platelet recovery was affected by exposure to alkylating agents (P=0.04), refractory disease (P=0.02) and AML (P=0.0001), but only the last two variables remained significant in Cox regression (P<0.01). Granulocyte engraftment was longer in CML (univariate, P=0.04) and in refractory disease (multivariate, P=0.02). In patients re-infused with >2 x 10(6)/CD34(+)/kg, the Cox model did not identify prognostic factors for haematopoietic recovery. CONCLUSION: Although mobilisation schedules and disease status influenced not only the yield of progenitor cells, but also the engraftment kinetics, the number of CD34(+) re-infused was the main predictor of haematopoietic recovery. While engraftment succeeded in most of the cases, the re infusion of >2 x 10(6)/CD34(+)/kg resulted in significantly shorter recovery times. PMID- 11920218 TI - Molecular characterization of four novel mutations causing factor VII deficiency. AB - INTRODUCTION: Hereditary deficiency of factor VII (FVII) is a rare coagulation defect. We previously studied the molecular basis of the FVII deficiency in Israeli patients and found that the majority of them bore the Ala244Val mutation. In the present study we further analysed FVII deficient patients. PATIENTS AND METHODS: Three patients with severe FVII deficiency (FVII activity < or =1%) and one with partial deficiency (25%) were studied. In all four patients, the FVII gene was amplified and sequenced. RESULTS: Four novel mutations have been identified: IVS 2+1G-->C Phe 24 deletion, Leu300Pro and Arg277His. Homozygosity for the IVS2+1G-->C mutation was lethal, whereas homozygosity for the Phe 24 deletion was accompanied by a severe bleeding tendency. FVII modeling showed that Phe 24 is located in the Gla domain. Both Arg 277 and Leu 300 are within the catalytic domain, although Arg 277 is also involved in tissue factor binding. CONCLUSION: We have analysed four mutations, two of which (IVS2+1G-->C, Phe 24 deletion) were associated with severe bleeding tendency in the homozygous state, facilitating prenatal diagnosis. Hypothetically, using FVII modeling, Arg 277 replacement by histidine may weaken the tissue factor, while deletion of Phe 24 and Leu300Pro mutation might be associated with abnormal folding of the Gla and catalytic domains, respectively. PMID- 11920219 TI - Biochemical and genetic defects underlying human congenital hypotransferrinemia. AB - INTRODUCTION: Human congenital hypotransferrinemia is a rare disorder characterized by the virtual absence of transferrin in the serum. No information on the causes of the disease is known. MATERIALS AND METHODS: Here we describe the identification of a new case, its treatment and the biochemical and genetic defects underlying the disorder. RESULTS: At diagnosis the patient had serum Tf levels equal to about 1% of the normal values. The treatment with plasma infusions each month allowed a good erythropoiesis and the prevention of iron overload with no need of red blood cell transfusions or iron chelators. In order to define the genetic basis of the disease, we performed a haplotype analysis of the Tf gene region in the 26 individuals forming the proband's family, and demonstrated that the genetic defect is located in the Tf gene and that it is inherited as a recessive trait. Protein analyses indicate that the proband serum contains two transferrin forms: one of 80 kD analogous to the normal one, and a smaller one of 50 kD, which may arise from a specific degradation or be the gene product of a modified allele. CONCLUSION: These data suggest the presence of two Tf alleles carrying genetic defects that cause two distinct abnormalities. One allele causes low expression of an apparently normal protein that probably allowed the survival of the patient in the first years of age. The other allele produces a modified Tf with different biochemical characteristics compared to the normal one. PMID- 11920220 TI - H RAS mutations in haematologically normal individuals. AB - INTRODUCTION: Point mutations in N, K and H RAS have been found in adverse haematological malignancies. The background frequency of RAS mutations in the normal population has yet to be determined. Here we report the results of a screen for RAS mutations from normal individuals. MATERIALS AND METHODS: DNA from peripheral blood or bone marrow from 115 haematologically normal individuals was screened for point mutations in N, K and H RAS, at amino acid positions 12, 13 and 61. The screening was done using polymerase chain reaction and oligonucleotide hybridisation and candidate mutations were subsequently confirmed by cloning and sequencing. RESULTS AND CONCLUSION: Point mutations were identified in DNA from two of the 115 individuals. Both mutations resulted in an amino acid substitution at position 12 in H RAS. Both individuals with detectable H RAS mutations remain haematologically normal. PMID- 11920221 TI - HLA class I-minor histocompatibility antigen tetramers select cytotoxic T cells with high avidity to the natural ligand. AB - INTRODUCTION: Cytotoxic T cells specific for the hematopoietic system-restricted minor histocompatibility antigens HA-1 and HA-2 are potential tools for the treatment of relapsed leukemia after minor histocompatibility antigen mismatched bone marrow transplantation. HA-1/HA-2-specific cytotoxic T cells with strong cytotoxic activity against HA-1/HA-2 positive target cells can be generated in vitro using HA-1 and HA-2 peptide-pulsed dendritic cells as antigen presenting cells. MATERIAL AND METHODS: We used HLA-A2 HA-1/HA-2 tetramers (HA-1(A2)/HA 2(A2) tetramers) to monitor the in vitro generation of HA-1- or HA-2-specific cytotoxic T cells. RESULTS: We show that the intensity of the tetramer-staining of the HA-1/HA-2-specific cytotoxic T cells strongly correlates with their capability to recognize mHag positive target cells. The bright tetramer-staining cytotoxic T cells lyse target cells expressing the natural ligand. The dim tetramer-staining cytotoxic T cells fail to lyse natural ligand positive target cells and lyse peptide-pulsed target cells only. The frequency of bright tetramer staining, high avidity minor histocompatibility antigen-specific CTLs increases significantly upon appropriate antigen-specific restimulations. CONCLUSION: Our results demonstrate that HLA class I-minor histocompatibility antigen tetramers are useful tools for monitoring and selection of high avidity HA-1- and HA-2 specific cytotoxic T cells for adoptive immunotherapy. PMID- 11920222 TI - Dose-finding study of valspodar (PSC 833) with daunorubicin and cytarabine to reverse multidrug resistance in elderly patients with previously untreated acute myeloid leukemia. AB - INTRODUCTION: This trial was designed to determine the maximum tolerated dose of intravenous daunorubicin (DNR) in combination with valspodar and to test the feasibility of P-glycoprotein modulation using valspodar in elderly patients with previously untreated acute myelogenous leukemia receiving standard induction chemotherapy. METHODS: Patients > or =60 years of age with previously untreated AML received valspodar (10 mg/kg/24 h by continuous intravenous infusion [CIV] on days 1-4 with a 2-mg/kg loading dose on day 1) in conjunction with two cycles of induction chemotherapy consisting of cytarabine (200 mg/m(2) CIV on days 1-7), and DNR (35 mg/m(2) [cohort 1] or 45 mg/m(2) [cohort 2] on days 1-3, intravenous bolus). Patients were assessed for dose-limiting toxicities (DLT), response rate, event-free and overall survival, and pharmacokinetics of valspodar and DNR. RESULTS: Valspodar was well tolerated at the lower DNR dose level (ie, 35 mg/m(2)) resulting in a 21% rate of DLT and only three toxic deaths. Treatment related mortality was unacceptably high at the 45 mg/m(2) DNR dose level. The complete response rate was 49% overall and similar in both cohorts. The median overall survival of patients was 333 days in cohort 1 compared to 98 days in cohort 2. At baseline, 70% of assessable patients were P-glycoprotein positive. CONCLUSION: Substantial inhibition of P-glycoprotein activity can be achieved in this patient population at clinically tolerable doses of valspodar and DNR. The maximum tolerated dose of DNR was established as 35 mg/m(2). This regimen is being further evaluated in phase III trials. PMID- 11920223 TI - Do anthracyclines have a role in the therapy of multiple myeloma? AB - INTRODUCTION: There is a great need for alternative treatments for patients with relapsing and refractory multiple myeloma. The new anthracycline idarubicin has the advantage of oral administration and has been suggested as part of new orally based chemotherapeutic combination regimens. The evidence of its own efficacy in this disorder is, however, insufficient. MATERIALS AND METHODS: In a multi-centre phase II study we administered oral idarubicin as a single drug in a dose of 10 mg/m(2) days 1-4 every four weeks to a total of 30 patients with relapsing (n=18) or refractory (n=12) multiple myeloma. RESULTS: Of 28 evaluable patients only one achieved a partial response of 15 months' duration. Other patients showed a very short minor response (n=1), "no change" (n=11) or progression (n=15) during the therapy. Toxicity was mostly mild and the drug was fairly well tolerated. None the less, half of the patients experienced WHO grade 3 and 4 toxicity in the form of granulocytopenia (n=10) or thrombocytopenia (n=4). CONCLUSION: Based on our experience and available data from three previously published reports, we consider idarubicin to have only a marginal effect in relapsing and refractory myeloma. A review of the literature on studies of anthracylines as single-agent therapy shows that only 5% of patients (19 out of 377) show a partial response. In our opinion the regular inclusion of an anthracycline in drug combinations for refractory myeloma should be reconsidered. PMID- 11920224 TI - In defense of Phase I trials. PMID- 11920227 TI - John Hughes Bennett, Rudolph Virchow... and Alfred Donne: the first description of leukemia. PMID- 11920229 TI - Tissue factor coagulation pathway and blood cells activation state in renal insufficiency. AB - INTRODUCTION: Atherosclerotic cardiovascular disease is the leading cause of the increased morbidity and mortality observed in uremic patients. Thrombosis is an important contributor to the evolution of atherosclerotic lesions. The physiologically-relevant blood clotting depends on binding of activated factor VII (FVIIa) to exposed tissue factor (TF) on activated/damaged cells. MATERIALS AND METHODS: A cross-sectional study was performed on three age- and sex-matched groups of individuals: one group of 50 patients on maintenance hemodialysis (D group), one of 50 patients with a non-dialysed renal insufficiency (ND group) and one of 50 healthy controls (HC group). We studied basal plasma concentrations of FVIIa, factor VII-related antigen (FVIIAg), soluble TF, tissue factor pathway inhibitor (TFPI), TF-dependent circulating monocytes procoagulant activity (TF dMPA), tissue factor-dependent plasma reactivity to activated protein C (TF-aPC), D-dimers (D-Di), and circulating markers of cellular activation/injury: soluble thrombomodulin (sTM), circulating microparticles (microP), soluble leukocyte, endothelial and platelet selectins (sL-selectin, sE-selectin, sP-selectin), soluble intercellular adhesion molecule 1 and vascular cell adhesion molecule 1 (sICAM-1 and sVCAM-1). Their variations induced, in hemodialysis patients, by a dialysis run were thereafter studied RESULTS: Values of FVIIa, FVIIa/FVIIAg ratio, sTF, TFPI, TF-dMPA, D-Di, sTM, microP, sL, sE and sP selectins, sICAM-1 and sVCAM-1 increased all along the hierarchy HC group/ND group/D group. Microparticles were mainly of platelet origin, to a lesser extent of monocyte origin. Dialysis induced an increase of FVIIa, sTF, TF-dMPA and circulating markers of cellular activation/injury. Strong correlations were observed between FVIIa/FVIIAg ratio and serum creatinine levels, sTF, TF-dMPA, sTM, sE-selectin, sVCAM-1. The TF-aPC was impaired in the ND and the D group, and the lower values were, in the D group, associated with antecedents of vascular access thrombosis. CONCLUSION: Renal insufficiency is associated to an activation of the tissue factor coagulation pathway, to a platelet, monocyte and endothelial activation/injury and to a deficient tissue-factor induced response to activated protein C which culminate in end-stage disease and are increased by hemodialysis runs. This contributes to linked coagulation and cellular conditions for an enhanced atherosclerosis progression. Due to the TF pathway activation, the therapeutic use of recombinant TFPI should be evaluated. PMID- 11920228 TI - Central role of leukemia-derived factors in the development of leukemia associated immune dysfunction. AB - INTRODUCTION: The underlying mechanism of tumor-associated T-cell dysfunction has been suggested to be due to an acquired abnormality in the T-cell receptor signaling complex, however, tumor-derived inhibitory factors acting on immune effectors could also explain the observed immunosuppression. Here, in a murine acute leukemia model, responses (proliferation and IL-2 secretion) to mitogens were suppressed in the early stages of leukemic progression due to a leukemia derived soluble factor(s). MATERIALS AND METHODS: Correlations between the development of immunosuppression and changes in expression of TCR signaling proteins (CD3-zeta, p56(lck), p59(fyn)), calcium mobilization, and total tyrosine kinase activity following TCR stimulation were examined. RESULTS: In contrast to reduced responses to mitogens seen as early as 1 week following injection of leukemic cells, signaling abnormalities were only seen in advanced disease (ie four weeks). The only significant alteration in signaling protein expression was loss of CD3-zeta for four weeks following the initiation of leukemia. Importantly, a direct role of a leukemia-derived soluble factor(s) in the loss of CD3-zeta could be shown in vitro by co-culturing splenocytes with leukemic cells separated by a transwell for seven to 10 days. Further studies demonstrated that the leukemia-derived factor(s) stimulated splenic macrophages to secrete a second soluble factor(s) that caused the loss of CD3-zeta. CONCLUSION: These data indicate that soluble leukemia-derived factor(s) have both immediate immunosuppressive actions and a later effect acting indirectly to induce intrinsic defects in T-cell signaling pathways via accessory cells. In some tumors, there is evidence that the soluble factor is Fas-ligand, which induces apoptosis in tumor-infiltrating T cells and the action of activated caspases cleave CD3-zeta. In this model, the soluble factor prevents apoptosis indicating a second mechanism is responsible for the loss of T-cell signaling proteins. PMID- 11920230 TI - Interferon-alpha protects Philadelphia-negative progenitors from exhaustion in chronic myeloid leukemia patients with cytogenetic response. AB - INTRODUCTION: Normal immature hematopoietic progenitors are relatively well preserved in most patients newly diagnosed with chronic myeloid leukemia, but tend to decline rapidly with time. Such exhaustion could reflect a suppressive effect of the Philadelphia positive clone expansion and/or be induced by Interferon-alpha treatment. MATERIALS AND METHODS: A total of 51 CML patients were classified into three groups. Newly diagnosed untreated patients were group A (n=30). Of the 21 treated individuals with Interferon-alpha, for at least 12 months, 15 showed no cytogenetic response (group B) while six showed persisting major/complete response (group C). Patients belonging to groups A and B were mobilized with chemotherapy plus G-CSF while patients of group C received a short course of G-CSF only. RESULTS: Patients responding to IFN-alpha (group C) showed comparable numbers of bone marrow Ph- long-term culture initiating cells to those of newly diagnosed individuals (group A): 8.5 (<1-65)/10(6) MNC vs 10.5 (<1-30), while non-responders had markedly lower numbers: <1 (<1-5). The amount of Ph- LTC IC collected was significantly lower in patients of group B 1.8 (0-325)x10(2)/kg than in patients of either group A 31.3 (0-952)x10(2)/kg (P<0.002) or group C 109 (8-259)x10(2)/kg (P<0.01). Interestingly, five patients of group B who had 100% Ph+ metaphases, but Ph- progenitors in their bone marrow, mobilized normal amounts of Ph(-) progenitors. CONCLUSION: These findings suggest that the decline of normal hematopoietic progenitors, currently observed in the majority of CML patients, is not induced by IFN-alpha treatment, but it is likely due to the expanding leukemic clone. They also indicate that normal hematopoietic reservoir is consistently preserved in patients given IFN-alpha early after diagnosis and achieving a stable cytogenetic response. PMID- 11920231 TI - Detection of CD55- and/or CD59-deficient red cell populations in patients with lymphoproliferative syndromes. AB - INTRODUCTION: Paroxysmal nocturnal hemoglobinuria is an acquired clonal stem cell disorder characterized by the decrease or absence of glycosylphosphatidylinositol anchored molecules from the surface of the affected cells, such as CD55 and CD59, resulting in chronic intravascular hemolysis, cytopenia and increased tendency to thrombosis. PNH-phenotype has been described in various hematological disorders, mainly in aplastic anemia and myelodysplastic syndromes, while it has been reported that complete deficiency of CD55 and CD59 has also been found in patients with lymphoproliferative syndromes, like non-Hodgkin's lymphomas. MATERIALS AND METHODS: The presence of CD55- and/or CD59-defective red cell populations was evaluated in 217 patients with lymphoproliferative syndromes. The study population included 87 patients with NHL, 55 with HD, 49 with CLL, 22 with ALL and four with hairy cell leukemia. One hundred and twenty-one healthy blood donors and seven patients with PNH were also studied as control groups. The sephacryl gel microtyping system was performed for the detection of CD55- and CD59-deficient red cell populations. Ham and sucrose lysis tests were also performed in all samples with CD55 or CD59 negative populations. RESULTS: Red cell populations deficient in both CD55 and CD59 molecules were detected in 9.2% of patients with lymphoproliferative syndromes (more often in ALL and nodular sclerosis type of HD) and in all PNH patients. CD55-deficient red cell populations were found in 8.7% of LPS patients (especially in low grade B-cell NHL), while CD59-deficient populations were found in only two patients with low grade B-cell NHL. CONCLUSION: These data indicate a possible association between paroxysmal nocturnal hemoglobinuria phenotype and lymphoproliferative syndromes, while further investigation is necessary to work out the mechanisms and the significance of the existence of this phenotype in these patients. PMID- 11920232 TI - Methionine synthase A2756G and methylenetetrahydrofolate reductase A1298C polymorphisms are not risk factors for idiopathic venous thromboembolism. AB - Hyperhomocysteinemia is a defined risk factor for venous thromboembolism (VTE). Several polymorphisms of genes encoding for enzymes acting in the remethylation pathway of homocysteine metabolism, ie, methionine synthase (MS) A2756G, methylenetetrahydrofolate reductase (MTHFR) C677T and MTHFR A1298C, can cause increased homocysteine levels particularly in patients with deficiencies of folic acid, vitamin B6, or B12 and hence be potential risk factors for VTE. Indeed, homozygous MTHFR C677T was shown to be a mild risk factor for VTE by some, but not by all, investigators. In this study, we assessed the risk exerted by MS A2756G and MTHFR A1298C in a cohort of patients with idiopathic venous thromboembolism. Homozygosities for MS A2756G and MTHFR A1298C were not found to be statistically significant risk factors for VTE. In addition, no interactions were observed among MS A2756G, MTHFR A1298C and MTHFR C677T in conferring a risk of VTE. PMID- 11920233 TI - Gp130 and ras mediated signaling in human plasma cell line INA-6: a cytokine regulated tumor model for plasmacytoma. AB - INTRODUCTION: Cytokines of the gp130-family, particularly interleukin(IL)-6, play a crucial role in the propagation of malignant plasma cells. MATERIALS AND METHODS: The role of IL-6 and other gp130-cytokines was studied in the human plasma cell line INA-6 in vitro and in INA-6 xenografts. The proliferative response to gp130-cytokines was evaluated and activated components of gp130 signaling pathways were identified by Western blotting and DNA binding studies. Specifically, expression of IL-6 and receptors for IL-6 and leukemia inhibitory factor were analysed by RT-PCR and ELISA. RESULTS: The plasma cell line INA-6 was cultured for several years remaining strictly dependent on exogenous IL-6. Other gp130-cytokines had no significant effect on INA-6 cell proliferation in vitro. Due to an activating mutation in the N-ras gene, mitogen-activated protein kinases (MAPK) were constitutively phosphorylated. In contrast, signal transducer and activator of transcription(STAT)-3 activation was dependent on stimulation with IL-6. Blocking of either one of these pathways resulted in a significant decrease of INA-6 cell proliferation. Remarkably, INA-6 xenografts did not require exogeneous IL-6 for proliferation in vivo. Instead, an autocrine IL-6 loop and, in certain tumor sublines, responsiveness to additional gp130-cytokines was induced during in vivo growth. CONCLUSION: Activation of the gp130 signal transducer is mandatory for INA-6 cell growth in vitro and in vivo. Both the MAPK and the Jak/STAT pathway are operative in malignant plasma cells and either one is essential for plasma cell growth. The INA-6 cell line provides a preclinical model to study growth regulation of human plasmacytoma cells and to evaluate novel therapeutic strategies. PMID- 11920234 TI - Detection of BCR/ABL rearrangements in adult acute lymphoblastic leukemia using a highly sensitive interphase fluorescence in situ hybridization method (D-FISH). AB - INTRODUCTION: One hundred-and-six adult cases of acute lymphoblastic leukemia were prospectively investigated using a highly sensitive interphase fluorescence in situ hybridization assay which utilizes DNA probes that detect a double BCR/ABL fusion signal (D-FISH) in cells carrying the t(9;22) to evaluate the reliability and specificity of this method for the detection of the Ph translocation. The results were compared with those obtained in the same cases by conventional cytogenetics and by reverse-transcription polymerase chain reaction. MATERIALS AND METHODS: The study was performed using DNA probes that span the common breakpoints of the t(9;22) translocation and that detect a double BCR/ABL fusion in cells carrying this karyotypic anomaly, one on the abnormal chromosome 9 and one on the Ph chromosome. RESULTS: Interphase D-FISH detected a high number of rearranged cases (22/106) compared to conventional cytogenetics (15/106) and RT-PCR (21/106). CONCLUSION: Interphase D-FISH emerges as a reliable, fast and relatively inexpensive tool for the detection of BCR/ABL rearrangements in adult ALL patients at diagnosis. It has a sensitivity clearly higher than conventional karyotyping and it may prove also superior to that of RT-PCR in cases with unusual BCR/ABL breakpoints. Our results suggest that D-FISH might be considered as the initial test for the diagnosis of Ph+ adult ALL. PMID- 11920235 TI - Hypothesis for generation of the unstable Hb Bucuresti (beta 42 Phe-->Leu) mutation. AB - INTRODUCTION: Unstable hemoglobin disorders are characterized by a congenital, mostly familial chronic hemolytic anemia with episodes of severe hemolysis during febrile illnesses. Usually, isopropanol and heat stability tests lead to the diagnosis which is confirmed by protein and gene sequencing. Generation of the mutations is still a subject of controversy. PATIENT, MATERIALS AND METHODS: We describe a 6-year-old Swiss child with congenital hemolytic anemia and a negative family history. Hemoglobin was studied by IEF, HPLC reverse phase chromatography, heat stability and isopropranol tests. DNA was sequenced in both coding and non coding strands. RESULTS: An unstable Hb was diagnosed on the basis of positive heat stability and isopropranol tests. The TTT-->TTG mutation at codon 42 corresponding to a Phe-->Leu substitution was found on DNA sequencing. Paternity was confirmed indicating that we are dealing with a new mutation. CONCLUSION: So far, three different mutations at codon 42 of the beta-chain, and two at the corresponding position of the alpha-chain have been described, all leading to a hemolytic anemia. These mutations can either represent random phenomena occurring at an important location in the heme pocket, or may be secondary to the two highly homologous zones present in this region. These homologous zones may indicate a hot spot for point mutations created by abnormal crossing over or formation of loops, and an imperfect DNA repair process. PMID- 11920236 TI - Prevention of postsplenectomy sepsis. PMID- 11920238 TI - Are we building a new Tower of Babel? PMID- 11920239 TI - p16/INK4a gene inactivation by hypermethylation is associated with aggressive variants of monoclonal gammopathies. AB - INTRODUCTION: A model of a stepwise malignant transformation has been proposed for the pathogenesis of monoclonal gammopathies. In this model, cell cycle regulators play a central role as a source of genetic events; particularly, p16/INK4a gene acts as a tumoral suppressor gene and, recently, inactivation of this gene through a methylation mechanism, has been observed in multiple myeloma patients. Under the diagnosis of monoclonal gammopathies there is a broad spectrum of disorders with very different outcomes, ranging from indolent courses, such as those of monoclonal gammopathy of undetermined significance, Waldestron macroglobulinemia and smoldering multiple myeloma, to aggressive diseases such as symptomatic MM and primary plasma cell leukemia. To the best of our knowledge, the activity of p16 gene has not been evaluated and compared in these different subtypes of monoclonal gammopathies. MATERIALS AND METHODS: The methylation status of the p16 gene was analysed in a group of 159 patients with monoclonal gammopathies (40 monoclonal gammopathy of uncertain significance, eight Waldenstrom Macroglobulinemia, eight smoldering multiple myeloma, 98 symptomatic multiple myeloma and five primary plasma cell leukemia) using three different assays (restriction enzymes and PCR or S-B and modification by sodium bisulphite). RESULTS: Forty-one of 98 MM patients (41.8%) as well as four of the five (80%) primary PCL patients showed methylation of the p16 gene, while none of the patients with monoclonal gammopathy of undetermined significance, Waldenstrom Macroglobulinemia or smoldering multiple myeloma displayed a methylation status. CONCLUSION: These findings suggest that the methylation of the p16 gene could be a relevant oncogenic event in the monoclonal gammopathies evolution being associated with the most aggressive forms. PMID- 11920240 TI - A minimal serpin promoter with high activity in haematopoietic progenitors and activated T cells. AB - INTRODUCTION: The serine protease inhibitor Serpin 2A is highly expressed in ex vivo bipotent granulocyte/macrophage progenitor cells and in cultured myeloid stem cells. The gene undergoes rapid down-regulation as these cells are induced to differentiate, and constitutive expression in cultured myeloid stem cells retards maturation. Serpin 2A is also expressed in T cells as a consequence of activation. We now report analysis of the upstream regulatory elements that control Serpin 2A transcription. MATERIALS AND METHODS: Using primer extension and rapid amplification of cDNA ends the transcription start site of the Serpin 2A gene was mapped, and a 1.2 Kb genomic upstream fragment cloned and sequenced. Promoter activity and protein binding of deletion and site-directed mutant constructs were analysed by transient transfection and by electrophoretic mobility shift assays. RESULTS: A minimal promoter fragment was identified with high activity dependent on NF-kappa and Moloney murine leukaemia enhancer factor LVa binding sites in both myeloid stem cells and activated T cells. NF-kappa was shown to be the main DNA binding protein in T cells, whereas that in haematopoietic stem cells appears to be novel. CONCLUSION: Serpin 2A promoter activity in T cells is due predominantly to NF-kappa binding to its consensus site. Activity in haematopoietic stem cells appears to be mediated by a novel protein, which recognises the NF-kappa consensus only in the context of flanking sequences. This concise regulatory element may be of potential value in gene therapeutic applications. PMID- 11920241 TI - Naturally-occurring anti-G-CSF antibodies produced by human cord blood B-cell lines infected with Epstein-Barr virus. AB - INTRODUCTION: Naturally occurring antibodies (auto-Abs) recognizing human granulocyte-colony stimulating factor were detected with high frequency in serum samples obtained from umbilical cord blood of newborns (12 of 65 samples screened) and maternal peripheral blood serum samples from women at the end of gestation (seven of 56 cases tested). The aim of this paper was to demonstrate that auto-Abs anti-G-CSF revealed in the blood of newborns were produced during foetal life. MATERIALS AND METHODS: Mononuclear cells from cord blood samples of different newborns containing high titer anti-G-CSF Abs were infected with Epstein-Barr virus in vitro, and EBV-immortalized B-cell lines were isolated and characterized for specific anti-G-CSF Ab production. RESULTS: Six different, unrelated cell lines of male origin which showed the presence of EBNA-2 antigen in the nucleus, displayed a B-cell phenotype (CD30+, CD5-, CD10-, HLA-DR+, CD19+, CD20+, CD23+, CD38+, CD25+), coexpressed low intensity sIgM and sIgD, and produced only IgM with prevailing lambda clonal restriction and anti-rhG-CSF Ab reactivity. The Ab specificity was proven against either glycosylated or unglycosylated G-CSF by saturable binding in direct enzyme-linked immunosorbent assays, by competition binding and Western immunoblotting assays. CONCLUSION: The secreted Abs did not affect the in vitro generation of granulocyte colonies by human normal adult haemopoietic progenitor cells in soft agar clonogenic assays, suggesting that these Abs were not neutralizing. PMID- 11920242 TI - A clinical, molecular and cytogenetic study of 12 cases of human herpesvirus 8 associated primary effusion lymphoma in HIV-infected patients. AB - INTRODUCTION: Primary effusion lymphoma is a rare type of B-cell lymphoproliferative disorder which is mainly observed in patients with HIV infection. Lymphomatous cells bridge features of immunoblastic and anaplastic cells with a non-B non-T phenotype and are characterized by the presence of the human herpesvirus 8 genome. We report on the retrospective analysis of 12 cases. PATIENTS AND METHODS: : Twelve HIV-infected patients with serous effusions containing large HHV8(+) lymphomatous cells were extensively evaluated to disclose associated visceral involvement. Clonality was assessed by IgH gene rearrangement PCR analysis (n = 11) or Southern blot (n = 1). EBV and HHV8 DNA sequences were detected by PCR analysis. Cytogenetics studies were performed in seven cases using RHG-banding. RESULTS: Extraserous localizations of lymphoma were present in six cases (50%): mediastinal (n = 2), mesenteric (n = 2), pancreatic (n = 1), and bone marrow involvement (n = 1). A monoclonal rearrangement of IgH genes was demonstrated in six cases, an oligoclonal pattern in one, whereas no clonality could be detected in five. High HHV8 copy numbers were demonstrated in all effusion fluids, with EBV-co-infection in all cases but one. Cytogenetic analysis displayed a complex karyotype in all cases without recurrent abnormalities. Eight patients have died. Three patients are in complete remission at 28, 53 and 55 months after high-dose chemotherapy (n = 1), cidofovir and alpha-interferon combination therapy (n = 1), and antiretroviral therapy alone (n = 1). CONCLUSION: The clinical and molecular pattern, as well as the response to therapy suggest that primary effusion lymphoma represents an heterogenous type of virus-induced B-cell lymphoproliferative disorder, sharing pathophysiological features with that induced by the Epstein-Barr virus and occurring in immunocompromised patients. PMID- 11920243 TI - Selective inactivation of Von Willebrand factor binding to glycoprotein IIb/IIIa and to inhibitor monoclonal antibody 9 by site-directed mutagenesis. AB - INTRODUCTION: The purpose of this study was to assess the requirement for the RGD sequence of von Willebrand factor (VWF) for its binding to the beta3 chain of integrins and the structural basis for the specificity of monoclonal antibody (MoAb) 9 which specifically binds to VWF and inhibits this interaction. MATERIAL AND METHODS: : Seven point mutations were introduced into VWF by site-directed mutagenesis. Mutated recombinant VWF were tested for their multimeric pattern and their ability to bind to purified GPIIb/IIIa, thrombin-activated platelets and MoAb 9. RESULTS: All recombinant VWF were fully multimerized. The conservative Arg 1744 to Lys substitution into the RGD sequence resulted in an 80% loss of function whereas the Arg to Ala change led to a total loss of function. The two substitutions however did not impair the binding to MoAb 9. In contrast Arg 1715 to Ala substitution had no effect on the binding to GPIIb/IIIa but the binding of the corresponding mutated recombinant VWF to MoAb 9 was strikingly decreased. CONCLUSION: Direct evidence of the role of the structure and the charge of Arg 1744 into the RGD sequence were established by changing Arg to Lys (KGD) and to Ala (AGD). Our results also demonstrate that Arg 1715 is not essential in the function but it is necessary for maintaining the conformation recognized by MoAb 9 specific for the GPIIb/IIIa-binding domain of VWF. PMID- 11920244 TI - Rapid in vivo induction of leukocyte tissue factor mRNA and protein synthesis following low dose endotoxin administration to rabbits. AB - INTRODUCTION: Disseminated intravascular coagulation in humans is frequently associated with Gram-negative bacterial sepsis. Therefore, to examine the role and time frame of the in vivo induction of tissue factor (TF) by bacterial endotoxin, a reverse transcription polymerase chain reaction and a solid-phase ELISA assay were developed to monitor the in vivo production in rabbits, of TF mRNA and TF antigen by peripheral blood leukocytes (PBL). METHODS: : Healthy rabbits were injected intravenously with either 1, 10 or 50 microg/kg of Salmonella endotoxin. Blood samples were obtained both before endotoxin administration and at various time points thereafter, up to 24 h. Some experiments were also done to determine whether all-trans retinoic acid would ameliorate the signs of the endotoxin-induced disseminated intravascular coagulation. RESULTS: PBL counts dropped significantly within 2 h of rabbits receiving the endotoxin, recovering to baseline levels by 24 h. Platelet counts decreased gradually over this same time frame. Fibrin deposition was noted in renal glomerular capillaries at 24 h. An increase (P<0.001) in PBL-associated TF mRNA levels was observed 2 h post-endotoxin (10 microg/kg, n = 8), followed by a gradual decline over the subsequent 24 h. The average increase in TF mRNA at 2 h was approximately 4.6-fold (P<0.001) over that seen at time 0. The amount of mononuclear cell associated TF antigen demonstrated a peak at 2 h post-endotoxin (10 microg/kg, n = 13), with levels approximately 9.6-fold greater than (P<0.001) baseline. Pre-treatment of rabbits with all-trans retinoic acid significantly (P<0.001) ameliorated the PBL-associated increase in TF mRNA and TF antigen levels. CONCLUSION: These results suggest that low dose endotoxin (10 microg/kg) faithfully reproduces the non-overt activation of coagulation observed in primates and human volunteers, supporting the hypothesis that TF expression is involved in the in vivo initiation and propagation of disseminated intravascular coagulation. Moreover, all-trans retinoic acid may be effective in modulating in vivo the TF transcription induced by endotoxin. PMID- 11920245 TI - Possible oxidative stress involvement in congenital dyserythropoietic anemia type 1. AB - INTRODUCTION: Congenital dyserythropoietic anemia type 1 (CDA1) patients may suffer from iron overload, associated with oxidative damage. The aim of this study was to evaluate possible involvement of oxidative stress in the pathogenesis of CDA1. STUDY DESIGN: : Blood samples from 10 children diagnosed as CDA1 patients from five Bedouin families, were studied. In this study, activities of superoxide dismutase and catalase were evaluated as well as methemoglobin, plasma total thiols, plasma total antioxidant capacity and glycerol lysis time. RESULTS: Normal values were found for superoxide dismutase, methemoglobin, trolox equivalent antioxidant capacity and total plasma thiols in CDA1 patients. However average catalase levels were significantly reduced (P<0.001) and glycerol lysis test was significantly prolonged (P<0.001). Ferritin levels, which were slightly increased in all patients, positively correlated with catalase values (r = 0.74, P = 0.022). CONCLUSION: Oxidative stress has not been proven in CDA1 pediatric patients. Some indications of oxidative damage exist, but it may not be directly related to the mechanism of anemia. PMID- 11920247 TI - Identification of a novel CBFB-MYH11 transcript: implications for RT-PCR diagnosis. AB - INTRODUCTION: The chromosome rearrangements inv(16)(p13q22) or t(16;16)(p13;q22) are present in approximately 10% of all cases with de novo acute myeloid leukemia and define a subgroup with a favorable prognosis. Both aberrations result in a CBFB-MYH11 fusion gene that can be detected by RT-PCR. PATIENTS AND METHODS: To date, a total of 10 different in-frame CBFB-MYH11 fusion transcripts have been identified. A newly described transcript can not be amplified with the commonly used PCR primers since the MYH11 junction is located outside the amplified region (MYH11 cDNA position 2134). RESULTS: We describe here a robust two-step RT-PCR assay that reliably detects all known CBFB-MYH11 transcripts types, including the new variant. CONCLUSION: Because all previously established RT-PCR protocols may miss the new CBFB-MYH11 transcript, we propose to use the improved RT-PCR approach described here for the reliable detection of all known CBFB-MYH11 fusion transcripts. PMID- 11920246 TI - How do sickle cells become dehydrated? PMID- 11920248 TI - Acute priapism in a patient with unstable hemoglobin Perth and Factor V Leiden under effective oral anticoagulant therapy. PMID- 11920249 TI - Is there a limit to the normality of blood products? PMID- 11920250 TI - We treat patients and we do not treat disease. PMID- 11920251 TI - How to limit the overabundance of health care administrators. PMID- 11920252 TI - High-dose idarubicin and busulphan as conditioning for autologous stem cell transplantation in acute myeloid leukemia: a feasibility study. AB - INTRODUCTION: Between 30 and 50% of patients with acute myeloid leukemia still relapse after autologous stem cell transplantation. We investigated the feasibility of a new conditioning regimen consisting of high dose IDA plus oral busulphan in patients undergoing autologous transplantation. MATERIALS AND METHODS: Patients (n = 13) were given three days continuous infusion IDA, followed by four days conventional dose oral busulphan as conditioning. Peripheral blood stem cells were used in all cases. Eleven patients were in CR1. Patients with t(8;21) and inv(16) as well as those with acute promyelocytic leukemia were excluded from the study. The median of CD34+ cells infused was 6.2 x 10(6)/l (2.6-16.1). RESULTS: No case of transplant-related mortality occurred. The median number of days to neutrophil (>0.5 x 10(9)/l) and platelet (>20 x 10(9)/l) recovery was 10 (7-21) and 20 (9-26), respectively. Patients needed a median of 3 platelet units (1-6) and 3 blood units (0-12), respectively. Left ventricular ejection fraction remained unmodified after ASCT. Twelve out of 13 patients (92%) had variable grade of mucositis (two grade 2, five grade 3 and five grade 4). Total parenteral nutrition was needed in nine patients (69%). After a median follow-up of 14 months from ASCT, 11 patients out of 13 (85%) are alive in continuous CR; the other two patients experienced relapse at 12 and 14 months. CONCLUSION: Our data demonstrate the feasibility of a conditioning regimen based on high-dose IDA plus Busulphan in AML. Results concerning antileukemic efficacy are promising, but need confirmation on larger series with longer follow-up. PMID- 11920253 TI - Decreased function of Fas in patients displaying delayed progression of HIV induced immune deficiency. AB - INTRODUCTION: In acquired immune deficiency syndrome patients, apoptosis of uninfected lymphocytes may contribute to development of immune deficiency. This process may involve recruitment of Fas by human immunodeficiency virus products. In line with this possibility, the viral envelope glycoprotein gp120 does not induce death of T cells from subjects with the autoimmune/lymphoproliferative syndrome displaying defective Fas function. This study evaluates the possibility that Fas function defects delay progression of HIV-induced immune deficiency. MATERIALS AND METHODS: The susceptibility to Fas-induced cell death was assessed on T cells from 18 'long-term non-progressor', four 'non-progressor', four 'progressor' asymptomatic HIV-1-infected, and nine AIDS patients using anti-Fas monoclonal antibodies. RESULTS: Fas-induced cell death was significantly lower in long-term non-progressors and non-progressors than in normal controls, progressors, and AIDS. The single-patient data showed that 9/18 long-term non progressors and 3/4 non-progressors, but no progressors or AIDS were resistant to Fas. Analysis of the uninfected parents of two long-term non-progressors displaying decreased Fas-function showed that the mother of one of them and the father of the other displayed the same Fas function defect as their children. Fusion of T cells from Fas-resistant individuals with a Fas-sensitive cell line gave rise to Fas-resistant hybrid lines not carrying HIV, which suggests that the resistant phenotype is due to molecules exerting a dominant negative effect on a normal Fas system. CONCLUSION: These data suggest that Fas-resistance in long term non-progressors may be due to inherited alterations of the Fas signaling pathway and may be a novel factor in delayed progression. PMID- 11920254 TI - No BCL-2 protein over expression but BCL-2/IgH rearrangements in B cells of patients with persistent polyclonal B-cell lymphocytosis. AB - INTRODUCTION: Persistent polyclonal B-cell lymphocytosis is a rare hematological disorder, characterized by a chronic, stable and absolute polyclonal lymphocytosis, the presence of binucleated lymphocytes, a polyclonal increase in serum IgM immunoglobulin and clonal cytogenetic abnormalities involving chromosome 3. For explaining the expansion of B-lymphocytes pool in PPBL, an association with cigarette smoking and/or chronic Epstein-Barr virus infection have been suggested but both hypotheses have been ruled out. MATERIALS AND METHODS: We studied the presence of BCL-2/IgH rearrangements in a series of eight PPBL patients (seven females and one male) by a nested polymerase chain reaction (PCR), targeting the Major Breakpoint Region in BCL-2 locus and we explored the BCL-2 protein expression by Western blot. RESULTS: We demonstrated: (a) the constant presence of BCL-2/IgH rearrangements in eight out of eight DNA samples, (b) multiple rearrangements in three out of eight cases and, (c) normal BCL-2 protein expression, as compared to BCL-2 level in B-lymphocytes from healthy population. CONCLUSION: Despite the presence of BCL-2/IgH rearrangements, the accumulation of B lymphocytes in PPBL is not related to an overexpression of BCL 2 protein. PMID- 11920255 TI - Identification of two subgroups of mantle cell leukemia with distinct clinical and biological features. AB - INTRODUCTION: Mantle cell leukemia (MCLeu) has been considered as a leukemic form of mantle cell lymphoma (MCL). However, the presence of certain features rarely observed in MCL, such as transformation to prolymphocytic leukemia (PLL) or indolent clinical course, suggests that MCLeu may represent a distinct disorder. METHODS: Seven cases of MCLeu with t(11;14)(q13;q32) and BCL1-IGH gene rearrangement were ascertained among 140 newly diagnosed chronic B-cell lymphoproliferative disorders with leukemic expression. Comparative genomic hybridization, FISH for specific gene loci, and immunological studies were preformed in them. RESULTS: In comparison with CLL, MCLeu cases had low immunological scores < or =2 with respect to B-CLL (P<0.0001). Expression of CD38 was absent in 43% of MCLeu and in 44% of B-CLL. Comparative genomic hybridization analysis identified genomic imbalances in 86% of MCLeu with a similar pattern than in MCL: gains of 3q, 8q involving MYC gene and 15q, and losses of 6q, 9p, 13q and 17p affecting P53 gene. Differently from MCL and CLL, genomic loss of 8p was frequently detected in MCLeu (83%). Although clinical presentation of MCLeu was indistinguishable from CLL, all patients but one had disease progression within three years. According to the immunologic and genomic profiles, two distinct subgroups of MCLeu were defined: one related to PLL, showing CD38-, deletion of P53, and MYC amplification and another which corresponds to a leukemic form of classical MCL, presenting with CD38+ and normal P53 and MYC status. CONCLUSION: MCLeu and MCL are closely related disorders, as they show similar genomic and molecular patterns. However, the deletion of the short arm of chromosome 8 may represent a specific marker for MCLeu. Two distinct subgroups of MCLeu may also be distinguished according to the immunologic and genomic cell profiles. PMID- 11920256 TI - Release of cytokines and soluble cytokine receptors after intravenous anti-D treatment in children with chronic thrombocytopenic purpura. AB - INTRODUCTION: Immunoglobulin anti-D administration is one of several methods used in treating children with chronic immune thrombocytopenic purpura. Fc receptor blockade of the reticuloendothelial cell system and of mononuclear phagocytes is an important mechanism of the action of anti-D in ITP. Recently other possible mechanisms by which anti-D works in ITP have been considered. METHODS: The aim of this study was to obtain a better understanding of the effect of anti-D administration on cytokine, soluble cytokine receptors and platelet count in children with chronic ITP and to determine the pre-treatment plasma cytokine profile in this group of patients. Eighteen children with chronic ITP were examined. In our study the impact of anti-D on the cytokine network was evaluated by analysing serum levels of IL-6, IL-8, tumor necrosis alpha and soluble TNF receptors I and II by the EASIA method before and 1, 3, 20 and 40 h then seven days and one month after anti-D infusion. RESULTS: Anti-D caused a significant increase in platelet count 20 h postinfusion in 10 out of 18 children, 96 h postinfusion in three children and 168 h postinfusion in one child. The mean duration of the response was four weeks. A significant and rapid increase in plasma levels of IL-6, IL-8 and TNF-alpha was seen within 1 to 20 h after anti-D infusion. This increase was accompanied by a prolonged elevation of soluble TNF receptors. There was a significant correlation between TNF-alpha and IL-8, IL-8 and IL-6, TNF-alpha and sTNFRI, and sTNF receptors I and II. CONCLUSION: These data demonstrate that anti-D infusion caused changes in the cytokine network and raises the question of whether the therapeutic effectiveness of anti-D is related to its immunomodulating properties. PMID- 11920257 TI - Invasive pulmonary fungal infection in hematologic patients: is resection effective? AB - INTRODUCTION: Invasive pulmonary aspergillosis carries a high mortality in neutropenic patients. Descriptive reports have shown early surgery to be feasible with acceptably low morbidity. The possible benefit of surgery has not been investigated in comparative studies. MATERIALS AND METHODS: In a retrospective cohort study encompassing a 15-year period, 54 (8%) of 697 consecutive patients with severe hematological disease required treatment for localized invasive pulmonary aspergillosis. Patients treated by antifungal drugs (medical group, n = 24) were compared to patients treated with additional early lung resection (surgical group, n = 30). Outcomes analysed were fungal progression and survival. RESULTS: Fungal progression at six months was 17% (95% CI 3-31) in the surgical group and 52% (95% CI 34-73) in the medical group (P = 0.005). Survival at six months was 70% (95% CI 53-87) in surgically and 42% (95% CI 24-62) in medically treated patients (P = 0.009). Adjusting for differences in WHO performance score (worse in the medical group) and duration of neutropenia (longer in the surgical group) in a multivariate analysis, a difference in relative risk of death (0.26; 95% CI 0.08-0.88; P = 0.03) remained in favor of surgery. CONCLUSION: In this retrospective study surgical intervention to treat invasive pulmonary fungal disease appeared to have a beneficial effect on the impact of disease control and survival. Differences in baseline characteristics of the two patient groups calls for cautious interpretation. A prospective randomized trial seems warranted. PMID- 11920258 TI - The specific enhancement of interferon alpha induced growth inhibition by BCR/ABL only occurs in multipotent cells. AB - INTRODUCTION: The causative oncogene in CML is the BCR/ABL protein tyrosine kinase. This stem cell disease is often treated with interferon alpha (IFN-alpha) which can initiate haematological and cytological remission which is associated with increased survival. There is however no clear indication of why CML cells are more responsive to IFN-alpha. MATERIALS AND METHODS: To establish if BCR/ABL increases the sensitivity of multipotent cells to IFN-alpha a temperature sensitive mutant of BCR/ABL was expressed in the multipotent haemopoietic stem cell line FDCP-Mix. The effect of IFN-alpha in terms of proliferation, induction of apoptosis, changes in cell cycle inhibitor proteins, and differentiation was assessed by [3H]thymidine incorporation, Annexin V and Western blot analysis. RESULTS: When the BCR/ABL tyrosine kinase was activated, the IFN-alpha-induced inhibition on the growth rate of the FDCP-Mix cell population was more marked than in control populations. The BCR/ABL-mediated effect was due to decreased rates of DNA synthesis. There was no IFN-alpha-mediated induction of apoptosis. This enhanced BCR/ABL mediated growth inhibition occurred over a range of growth factor concentrations and was independent of changes in p21(Cip1) and p27(Kip) levels. When FDCP-Mix cells were induced to differentiate into mature macrophages and neutrophils in the presence of IFN-alpha, there was increased sensitivity to IFN-alpha that was independent of BCR/ABL activity. CONCLUSION: BCR/ABL PTK expression in this primitive multipotent haematopoietic cell line results in an enhanced response to IFN-alpha. In contrast, the more mature myeloid progenitor cells are equally responsive to this growth inhibitor. This data may explain some of the clinical effects of IFN-alpha. PMID- 11920259 TI - Stem cell transplantation for chronic myeloid leukaemia: the role of infused marrow cell dose. AB - INTRODUCTION: Allogeneic stem cell transplantation is a potentially curative option for patients with CML. The optimal donor is an HLA-identical sibling but transplants using unrelated volunteers can also be successful. The factors influencing survival after allogeneic SCT for CML are reasonably well defined. Recently however, the Seattle group have emphasised the influence of a high marrow cell dose on outcome following volunteer unrelated donor SCT for high risk acute leukaemia. MATERIALS AND METHODS: We have sought to define factors impacting on transplant related mortality (TRM) in a population of CML patients after allografting with matched sibling or alternative stem cell donors at a single centre over a 20-year period, with emphasis on infused marrow cell dose. Factors entered into a multivariate analysis were: recipient age, recipient CMV serostatus, disease phase, donor sex, cell dose and frequency of CTLP reactivity. RESULTS: In multivariate analysis four factors had an adverse effect on TRM when using a VUD: low marrow cell dose (<3.65 x 10(8) TNC/kg, relative risk 2.05, CI 1.08-3.90, P = 0.029), late disease phase (relative risk 1.68, CI 1.03-2.74, P = 0.038), patient CMV seropositivity (relative risk 1.98, CI 1.25-3.13, P = 0.004) and high frequency of CTLP (relative risk 1.93, CI 1.18-3.13, P = 0.008). For HLA identical sibling donor transplants the only factor that adversely impacted on TRM was late disease phase (P = 0.0004 in univariate analysis). CONCLUSION: High infused cell dose is a new modifiable factor associated with reduced TRM following allogeneic SCT using an unrelated donor for the treatment of CML. The data support the recommendation that bone marrow harvest teams should aim to collect the highest possible number of nucleated cells for recipients of unrelated donor transplants. PMID- 11920260 TI - Increased conventional chemotherapy does not improve survival in multiple myeloma: long-term results of two PETHEMA trials including 914 patients. AB - BACKGROUND: Melphalan and prednisone (MP) has been the standard treatment for multiple myeloma (MM) for the last 30 years. Combination chemotherapy at conventional doses has not shown a significant prolongation of survival when compared to MP. There are few data comparing conventional chemotherapy at standard doses with conventional treatment at higher doses. We present the long term outcome of 914 patients from two randomized trials comparing three different dose intensity regimens. METHODS: From 1 January, 1985 to 31 December, 1989, 487 patients were randomized between MP (melphalan 9 mg/m(2) p.o. and prednisone 60 mg/m(2) days 1-4) and alternating VCMP (vincristine 1 mg i.v. on day 1, cyclophosphamide 500 mg/m(2) i.v. on day 1, melphalan 6 mg/m(2) p.o. on days 1-4, and prednisone 60 mg/m(2) on days 1-4) and VBAP (vincristine 1 mg i.v. on day 1, BCNU and doxorubicin 30 mg/m(2) i.v. each on day 1, and prednisone 60 mg/m(2) on days 1-4). From 1 January, 1990 to 31 May, 1994, 427 patients were randomized between VCMP/VBAP at the above detailed doses (VCMP/VBAP 'SD') and the same regimen increasing the doses of cyclophosphamide and doxorubicin from 500 to 1200 mg/m(2) and from 30 to 50 mg/m(2), respectively (VCMP/VBAP 'HD'). RESULTS: Increasing dose intensity produced a significantly higher partial response rate (31% vs 45% vs 51% for MP, VCMP/VBAP 'SD', and VCMP/VBAP 'HD', respectively; P < 0.01). However, a significantly early death rate was observed in the HD arm (7.7, 7.5 and 12.1% for MP, VCMP/VBAP 'SD', and VCMP/VBAP 'HD', respectively; P = 0.05). Median duration of response (20 vs 18 vs 19 months for MP, VCMP/VBAP 'SD', and VCMP/VBAP 'HD', respectively; P = NS) and median survival (25 vs 31 vs 29 months for MP, VCMP/VBAP 'SD', and VCMP/VBAP 'HD', respectively; P = NS) were similar in the three groups. MP produced a higher degree of thrombocytopenia than combination chemotherapy at standard (P = 0.002) or high dose (P = 0.01), this leading to a significantly higher dose reduction in the MP arm (P < 0.001 and P = 0.003 for VCMP/VBAP 'SD' and VCMP/VBAP 'HD', respectively). CONCLUSION: In these trials the response rate significantly correlated with the regimen intensity. However, no significant differences in response duration and survival were found. This highlights the limited role of conventional chemotherapy in MM and the need for further trials, aimed at determining the impact of new treatment approaches such as high-dose therapy/autotransplantation. PMID- 11920262 TI - Is there a role for epigenetic factors in the pathogenesis of epidermotropic cutaneous T-cell lymphomas (mycosis fungoides and Sezary syndrome)? PMID- 11920263 TI - One moral, many ethics. PMID- 11920261 TI - Outcome is not improved by the use of alternating chemotherapy in elderly patients with aggressive lymphoma. AB - INTRODUCTION: A prospective randomised study involving 810 elderly patients was conducted in an attempt to compare alternating chemotherapy with conventional first-line chemotherapy in aggressive non-Hodgkin's lymphoma in order to improve prognosis with an acceptable toxicity for elderly patients. PATIENTS AND METHODS: Patients included were 55-69 years old and had at least one adverse prognostic factor. Patients were treated either with ACVBP followed by consolidation (n = 396) or with an alternating regimen (n = 414). This regimen was an association of active drugs in NHL relapsing patients, alternating VIMMM with ACVBP for induction and alternation of VIM and ACVM in consolidation. Eight hundred and sixty-six patients were randomised. After histological review, 810 patients met the inclusion criteria: 396 in arm A, 414 in arm B. RESULTS: The complete response rate after induction was superior for conventional first-line therapy (58.5% vs 48%, P = 0.003) but at the end of treatment, the CR rate was not statistically different (52% vs 48%, P = 0.19). Conventional chemotherapy had a better five-year event-free survival than alternating regimen (33% (95% CI: 30 36%) vs 28% (95% CI: 26-30%), P = 0.0289) but overall survival was not statistically different (40% (CI 95% 38-42%) vs 36% (CI 95% 34-38%), P = 0.068). In this elderly high risk population, the toxicity was very high: 19% in arm A and 26% in arm B died during treatment. CONCLUSION: Alternating regimen did not improve outcome, was less efficient and more toxic. PMID- 11920264 TI - Von Willebrand factor-cleaving protease activity and proteolysis of von Willebrand factor in bone marrow transplant-associated thrombotic microangiopathy. AB - INTRODUCTION: Thrombotic microangiopathy (TM) of the fulminant type occurring in patients following bone marrow transplant (BMT) has clinical manifestations that are similar to thrombotic thrombocytopenic purpura (TTP) and hemolytic uremic syndrome, but the outcome is generally fatal despite conventional therapy. Idiopathic acquired TTP has been associated with IgG inhibitors to the cleaving protease of von Willebrand factor (vWF) in plasma. In this study, we investigated the role of the vWF protease and vWF proteolysis in the pathogenesis of BMT associated TM of the fulminant type. METHODS: vWF antigen level, vWF multimeric pattern, and vWF metalloprotease activity were investigated in the plasma samples of six consecutive patients with acute BMT-associated TM. Histologic and immunohistochemical studies were also performed on autopsy kidney specimens from four of the patients. All six patients had the fulminant type of the disorder with a fatal outcome and none of the patients responded to plasma infusion. RESULTS: The vWF-cleaving protease activity in plasma was normal in all patients. However, analysis of the vWF multimeric pattern showed a decrease of high molecular weight multimers. The decrease of large multimers may be caused by vWF platelet binding as well as shear enhanced proteolysis of vWF. In the four patients who had an autopsy, a pattern of arteriolar thrombosis, distinct from that of TTP, was detected in the kidneys. CONCLUSION: These findings suggest that BMT-associated TM of the fulminant type is a heterogeneous process and distinct from TTP in pathogenesis. Analysis of vWF protease and vWF multimeric distribution are valuable tools in making the distinction between BMT-associated TM and TTP. PMID- 11920265 TI - Variability in B-cell antigen expression: implications for the treatment of B cell lymphomas and leukemias with monoclonal antibodies. AB - OBJECTIVES: Antigen expression intensity is becoming important for decision making in relation to monoclonal antibody therapy. By quantifying CD20, CD22 and CD52 expression on chronic lymphocytic leukemia and normal (control) B cells, over time. The effect of Interleukin-4 therapy on CD20 antigen intensity on B-CLL cells in vivo was also determined. METHODS: Lymphocytes were purified at weeks 0, 4 and 8 from five B-CLL patients, five healthy volunteers and seven B-CLL patients receiving IL-4 therapy. The number of antigen receptor sites was calculated in molecules of equivalent soluble fluorochrome using flow cytometry. RESULTS: The mean number of CD20 receptors at baseline was significantly lower on B-CLL cells compared to normal B cells (8160 vs 87 046; P<0.0001). Similar results were obtained for CD22 (8630 vs 27 647; P<0.01), but not for CD52 (371 303 vs 409 484; P = 0.54). When soluble fluorochrome values at weeks 4 and 8 were analysed as change in per cent from baseline (delta%), there was <10 delta% variability in CD20 expression on control B cells, but considerable variability (22.5-67.5 delta%) on B-CLL cells. Expression of CD22 in CLL and control B cells varied by <15 delta%. CD52 on CLL B cells showed slightly greater variability (+/ 35 delta%) than that of CD22 (+/-15delta%), but less than that of CD20. IL-4 therapy did not consistently increase the CD20 expression on B-CLL cells in vivo. CONCLUSION: Our data confirm differences in intensity between different target antigens on B-CLL cells, and draws attention to the fact that a substantial variability may occur over time, which may influence clinical decision-making. Caution must be taken when interpreting in vitro results on cytokine-mediated receptor intensity up-regulation. PMID- 11920266 TI - Stem cell growth factor: in situ hybridization analysis on the gene expression, molecular characterization and in vitro proliferative activity of a recombinant preparation on primitive hematopoietic progenitor cells. AB - INTRODUCTION: In situ hybridization of whole mouse fetuses and their tibias with a stem cell growth factor (SCGF) antisense probe demonstrated specific expression of SCGF mRNA around skeletal tissues, particularly in bone marrow cells, proliferating chondrocytes, the perichondrium and periosteum, but little expression in resting or hypertrophic chondrocytes. METHODS: Recombinant human (rh) SCGF-alpha was purified from a conditioned medium of SCGF-alpha gene transfected CHO cells. The molecular mass of rhSCGF-alpha, 45 kDa, was shifted down to 40 kDa by digestion with endo-O-glycosidase and sialidase, suggesting O glycosylation of rhSCGF-alpha with sialic acids. RESULTS: For human bone marrow CD34+Lin- cells, rhSCGF-alpha alone did not stimulate colony-formation, but small cluster-formation (10.3 +/- 2.5/1 x 10(3) CD34+Lin- cells). It promoted growth of erythroid and granulocyte/macrophage (GM) colonies in the primary culture with erythropoietin and GM colony-stimulating factor (CSF) or G-CSF, respectively, and further supported GM progenitor cells in a short-term liquid culture. In contrast, rhSCGF-alpha suppressed stem cell factor (SCF)-stimulated erythroid bursts, indicating some competitive interaction between SCGF and SCF. rhSCGF alpha was synergistic with interleukin-3 and the flt3 ligand to enhance GM colony growth, but not synergistic with those inducing ex vivo expansion of GM progenitor cells. CONCLUSION: SCGF is selectively produced by osseous and hematopoietic stromal cells, and can mediate their proliferative activity on primitive hematopoietic progenitor cells. PMID- 11920267 TI - The bioavailability of oral fludarabine phosphate is unaffected by food. AB - INTRODUCTION: A prospective, open and randomized, two-way crossover study was conducted to evaluate the pharmacokinetics and bioavailability of oral fludarabine phosphate when taken on a full versus an empty stomach. The effectiveness of therapy was also assessed after two cycles of treatment, four weeks apart MATERIALS AND METHODS: Patients with chronic lymphocytic leukemia or low-grade non-Hodgkin's lymphoma were randomly assigned to two groups, both of which received two cycles of treatment with 90 mg of oral fludarabine phosphate administered when either fed or fasted. Patients in Group 1 (n = 8) received oral treatment on a full stomach for the first cycle then on a fasted stomach for the second, while those in Group 2 (n = 10) received their treatment in the reverse sequence. Oral fludarabine phosphate was administered on the first day of the two study cycles and intravenous fludarabine phosphate was administered on days 3-6. RESULTS AND CONCLUSION: Of 22 patients recruited, 18 (CLL n = 10; NHL n = 8) were eligible for efficacy and safety evaluation, and 16 for bioavailability and pharmacokinetic analyses. The response to oral 2-F-ara-AMP was rapid: by two treatment cycles, 12 out of 18 patients (66.7%) had achieved partial response. Of the six patients who did not respond, five patients (27.7%) had stable disease. There was no notable difference in the rate of response between patients with B CLL and lg-NHL. There was a marginal increase in total systemic availability of fludarabine phosphate when administered orally on a fed stomach (2-F-ara-A AUC((0 24 h)) = 3.28 +/- 1.48 microg.h/ml) compared to a fasted stomach (2-F-ara-A AUC((0-24 h)) = 3.05 +/- 1.56 microg.h/ml). Time to peak plasma concentration was slightly extended by the presence of food (2.2 +/- 1.0 versus 1.3 +/- 0.74 h) but the terminal half-life was unaffected. The minor differences in the pharmacokinetics of oral fludarabine phosphate when taken after food were not statistically significantly different and seem unlikely to be clinically relevant. The efficacy and safety data closely paralleled previous experience with the intravenous formulation. PMID- 11920268 TI - Somatostatin modulates G-CSF-induced but not interleukin-3-induced proliferative responses in myeloid 32D cells via activation of somatostatin receptor subtype 2. AB - INTRODUCTION: Somatostatin, originally identified as a peptide involved in neurotransmission, functions as an inhibitor of multiple cellular responses, including hormonal secretion and proliferation. Somatostatin acts through activation of G-protein-coupled receptors of which five subtypes have been identified. We have recently established that human CD34/c-kit expressing hematopoietic progenitors and acute myeloid leukemia (AML) cells exclusively express SSTR2. A major mechanism implicated in the antiproliferative action of somatostatin involves activation of the SH2 domain-containing protein tyrosine phosphatase SHP-1. While 0.1-1 x 10(-9) M of somatostatin, or its synthetic stable analog octreotide, can inhibit G-CSF-induced proliferation of AML cells, little or no effects are seen on GM-CSF- or IL-3-induced responses. MATERIALS AND METHODS: To study the mechanisms underlying the antiproliferative responses of myeloblasts to somatostatin, clones of the IL-3-dependent murine cell line 32D that stably express SSTR2 and G-CSF receptors were generated. RESULTS: Similar to AML cells, octreotide inhibited G-CSF-induced but not IL-3-induced proliferative responses of 32D[G-CSF-R/SSTR2] cells. Somatostatin induced SHP-1 activity and inhibited G-CSF-induced, but not IL-3-induced, activation of the signal transducer and activator of transcription proteins STAT3 and STAT5. CONCLUSION: Based on these data and previous results, we propose a model in which recruitment and activation of the tyrosine phosphatase SHP-1 by SSTR2 is involved in the selective negative action of somatostatin on G-CSF-R signaling. PMID- 11920269 TI - Feasibility and clinical significance of real-time quantitative RT-PCR assay of PML-RARalpha fusion transcript in patients with acute promyelocytic leukemia. AB - INTRODUCTION: To study the relationship between the expression level of the PML RARalpha fusion transcripts and the clinical status and efficiency of the therapy in acute promyelocytic leukemia (APL) patients, we applied a very sensitive and specific real-time Reverse Transcription Polymerase Chain Reaction (RT-PCR) system to quantify the dose of PML-RARalpha fusion transcripts in a series of APL patients at distinct disease stages. MATERIALS AND METHODS: A total of 31 APL patients (19 males and 12 females; aged from 8 to 74 years) from eight hospitals in Shanghai were analysed. Real-time Quantitative RT-PCR was used to measure the normalized dose (DoseN) of PML-RARalpha fusion transcripts. RESULTS: A wide range of PML-RARalpha DoseN above 1 x 10(3) was noted in 25 newly diagnosed patients. PML-RARalpha DoseN was significantly decreased after remission induction with ATRA, ATRA/chemotherapy or As2O3 and further reduced after consolidation. The fact that all patients with long disease free survival had a constantly low PML RARalpha DoseN below 2 x 10(2) and a higher level predicted impending relapse suggests that this value could serve as a 'threshold' for molecular remission. PML-RARalpha DoseN was also of prognostic value in a group of relapsed patients, since good response to As2O3 reinduction was accompanied by a remarkable reduction of fusion transcript level, whereas patients with high PML-RARalpha Dose(N) after the second CR tended to relapse again rapidly. CONCLUSION: These results confirm that real-time RT-PCR assay for PML-RARalpha transcripts in APL patients is useful in reflecting leukemic burden, assessing response to treatment and indicating the ultimate clinical outcome or curability of disease. PMID- 11920270 TI - Assessing the Cox model assumption as a statistical tool for classifying lymphomas. AB - INTRODUCTION: The Cox model is widely used in medical research for comparing survival. Lymphomas might exhibit important differences in long-term cure rate despite a similar survival. METHODS: Using log-rank test, we compared event-free survival (EFS), and the survival of 64 patients with mantle cell lymphoma (MCL), 525 patients with follicular lymphoma, and 1136 patients with diffuse centroblastic lymphoma (CB). RESULTS: Although EFS and survival of MCL were significantly shorter than those of follicular lymphoma, checking the validity of the proportional hazards assumption shows that the distribution of rates of events and deaths over time did not differ in MCL and follicular lymphoma. In contrast, the ratios of hazards (events and deaths rates) did not remain constant over time in MCL and CB, because of a decrease in late events and deaths rates in the latter histological type. CONCLUSION: Checking the validity of the Cox model hypothesis might be a useful tool for assessing long-term cure rate in seldom lymphoma subtypes. Despite a short overall survival, MCL should not be considered to be an aggressive lymphoma, in which available chemotherapy may cure a subset of patients. PMID- 11920271 TI - Total deficiency of specific von Willebrand factor-cleaving protease and recovery following plasma therapy in one patient with hemolytic-uremic syndrome. PMID- 11920272 TI - Prognostic relevance of soluble CD23 levels in CLL. PMID- 11920273 TI - How to compensate for the expense of new drugs. PMID- 11920274 TI - Acquired von Willebrand syndrome: an important bleeding complication to be considered in patients with lymphoproliferative and myeloproliferative disorders. PMID- 11920275 TI - Female donors influence transplant-related mortality and relapse incidence in male recipients of sibling blood and marrow transplants. AB - INTRODUCTION: The male H-Y antigen is recognised as a minor histocompatibility antigen with debatable relevance and has not been substantiated thus far in clinical solid organ transplantation. An increase in transplant-related mortality in male recipients of female stem cells has been recognised and attributed to graft vs host effect against H-Y; in contrast, decreased incidence of relapse, a graft vs leukaemia effect has not yet been described. METHODS: To detect potentially small but significant differences, we performed an analysis in a highly homogeneous group. We have analysed 782 patients, 438 males and 344 females, who were transplanted from HLA-identical female donors for CML in 1st chronic phase between 1989 and 1997. The risk of transplant related mortality (TRM) and relapse incidence (RI) were compared for male and female recipients over three successive time periods, zero to three months, three months to two years and two to five years post-transplant. Both groups were comparable for known risk factors prior to transplant. RESULTS: The cumulative risk of TRM at five years was significantly higher in males (42%, s.e.=3.6) than in females (27%, s.e.=2.8; P=0.02) with no overall effect on risk of relapse. Within the three specified time intervals post-transplantation, the number of events of TRM and RI in the two groups of recipients diverged over time. There was no difference within the first time interval (zero to three months) for both events. TRM was higher in male recipients as manifested from three months onwards over the whole observation period of five years (P=0.02). Risk of relapse was significantly reduced in male patients; this difference became manifest only beyond two years (P=0.01). CONCLUSION: These data confirm the importance of male gender for both, TRM and RI: male recipients of female blood or marrow grafts are at risk of enhanced procedure related mortality, eg GvHD, but benefit from reduced incidence of disease recurrence, hence a GvL effect. GvL in this setting needs more time than GvHD. These data give indirect but convincing evidence for a clinical relevance of H-Y as a minor histocompatibility antigen in humans. PMID- 11920276 TI - High molecular mass kininogen inhibits cathepsin G-induced platelet activation by forming a complex with cathepsin G. AB - INTRODUCTION: Preliminary studies have shown that high molecular mass kininogen (HK) inhibits cathepsin G-induced platelet activation. However, the potential mechanism underlying this inhibitory effect remains to be elucidated. MATERIALS AND METHODS: Suspensions of washed and gel-filtered platelets were used in radioligand binding and aggregation studies. The amidolytic activity of cathepsin G was measured using specific chromogenic substrate. Western blot technique was utilised to explore the potential complex formation between cathepsin G and HK. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis was used to analyse the cleavage products of HK. RESULTS: At a concentration of 1 microM, HK completely blocked cathepsin G-induced platelet shape change and secretion of ATP. HK inhibited cathepsin G-induced platelet aggregation in a concentration-dependent manner with an IC(50) of 0.48 microM. Moreover, HK was found to inhibit binding of (125)I-cathepsin G to gel-filtered platelets. (125)I-cathepsin G forms a complex with HK. The complex formation did not affect the amidolytic activity of cathepsin G. HK was proteolysed upon interaction with cathepsin G. CONCLUSION: Our results show that high molecular mass kininogen down-regulates cathepsin G induced platelet activation by forming a complex with cathepsin G and thus prevents binding of cathepsin G to platelets. These kininogen-cathepsin G interactions may be potential targets for pharmacological intervention. PMID- 11920277 TI - Tumor necrosis factor alpha release is a major biological event associated with rituximab treatment. AB - INTRODUCTION: In patients with low-grade non-Hodgkin's lymphoma, rituximab (MabThera) produces infusion-related toxicity, including fever, rigors, and chills in greater than 50% of those treated. The majority of these reactions are grade 1 or 2. MATERIALS AND METHODS: In the GELA study LNH98-5, a total of 400 elderly patients with previously untreated diffuse large B-cell lymphoma were randomized to treatment with CHOP or with rituximab plus CHOP (R-CHOP). In a detailed investigation of biological events which may be associated with adverse reactions specific to rituximab infusion, a subgroup of 55 patients (26 in the CHOP group and 29 in the R-CHOP group) were selected for measurement of several biological parameters at baseline and at 1, 4 and 8 h (H1, H4 and H8, respectively) after commencing therapy. For 27 patients, measurements included cytokine and complement levels. RESULTS: Baseline demographic and disease characteristics were similar for patients in both treatment groups. Compared with the CHOP treatment group, patients in the R-CHOP group had significantly higher post-treatment changes in neutrophil, lymphocyte, and monocyte counts, LDH levels, C3a levels, and TNF-alpha levels. In the R-CHOP group, neutrophil levels increased at H4 (P<0.05), lymphocyte levels decreased at H1 (P<0.05), H4 (P<0.001) and H8 (P<0.05), monocytes levels decreased at H1 (P<0.01), LDH levels increased at H4 (P<0.05) and H8 (P<0.01), and C3a decreased at H1 (P<0.01). The most statistically significant changes were observed for TNF-alpha levels: Mean values of TNF-alpha increased more than 250% at H1 and H4 and were still increased by 170% at H8 (P<0.001 at all timepoints). Since only six of the 55 evaluated patients had severe adverse events, it was not possible to correlate severe toxicity with these biological variations. CONCLUSION: This analysis demonstrates that rituximab infusion was rapidly followed by activation of complement, B-lymphocyte cytolysis, and TNF-alpha release. PMID- 11920278 TI - Down-stream regions of the POZ-domain influence the interaction of the t(11;17) associated PLZF/RARalpha fusion protein with the histone-deacetylase recruiting co-repressor complex. AB - INTRODUCTION: Acute promyelocytic leukemia (APL) patients with t(15;17)(PML/RARalpha positive) achieve remission upon t-RA treatment, whereas patients with t(11;17)(PLZF/RARalpha positive) do not. Both APL translocation products bind to the histone deacetylase (HD)-recruiting nuclear co-repressor complex (HD-NCR) in a ligand-dependent manner through their RARalpha portion. Differently to PML/RARalpha, PLZF/RARalpha also binds the HD-NCR in a ligand independent manner through the PLZF portion of the fusion protein (PLZF#), which seems to be crucial for the t-RA resistance of t(11;17) APL patients. MATERIALS AND METHODS: The t-RA sensitivity of U937 cells was tested by the nitro-blue tetrazolium reduction (NBT) assay and by analysis of t-RA-induced type II transglutaminase activity. The interaction between HD-NCR and PLZF/RARalpha was investigated by in vitro binding assays. RESULTS: (i) Deletions in PLZF# convert PLZF/RARalpha from a repressor to an activator of t-RA response in U937 cells; (ii) the effect of PLZF/RARalpha on t-RA-signaling is regulated by the POZ-domain and its down-stream regions of PLZF#; (iii) there are additional binding sites for HD-NCR in PLZF# and (iv) PLZF# not only directly binds but also regulates the binding of PLZF/RARalpha to the HD-NCR. CONCLUSIONS: At least two different mechanisms responsible for the aberrant recruitment of HD-NCR complexes by PLZF# are regulating the different t-RA-sensitivity of the PLZF/RARalpha and PML/RARalpha positive APL blasts: one is related to the direct binding of the different members of the HD-NCR complex to PLZF#; the other is an enforcing effect of PLZF# on the affinity of the PLZF/RARalpha fusion protein to the HD-NCR complex. PMID- 11920279 TI - Response to low-dose oral methotrexate and prednisone in two patients with angio immunoblastic lymphadenopathy-type T-cell lymphoma. AB - INTRODUCTION: AILD-type T-cell lymphoma is characterized by very poor prognosis in most patients and the response rate to conventional chemotherapy is unsatisfactory. MATERIALS AND METHODS: Two patients (a 65 year old female and a 67 year old male) with AILD-type lymphoma who did not respond to conventional treatment with steroids or aggressive chemotherapy were treated with Methotrexate and Prednisone. Both patients received a weekly dose of MTX (10 mg/m(2)) that was administered orally in combination with PDN at an initial dose (15 mg/day), given on a daily basis. RESULTS: Both patients responded rapidly showing marked improvement with no major side effects. Complete clinical remission was recorded in the two patients who were treated with this combination after conventional chemotherapy had failed to produce any improvement. CONCLUSION: Our observations in two patients with refractory/relapsed AILD-type lymphoma who were given low dose oral MTX as salvage treatment, suggest that this agent has immunosuppressive effects that can be beneficial for treating patients with AILD-type T-cell lymphoma. Pilot clinical trials are needed to verify its efficacy in this setting. PMID- 11920280 TI - Comparison of four serum-free, cytokine-free media for analysis of endogenous erythroid colony growth in polycythemia vera and essential thrombocythemia. AB - INTRODUCTION: The assay of endogenous erythroid colony formation (EEC), a characteristic of polycythemia vera and essential thrombocythemia, is not standardized. In this multicentric study, we tested four semisolid, serum-free, cytokine-free media based on either methylcellulose (M1, M2) or collagen (C1, C2) commercialized for the EEC assay. MATERIALS AND METHODS: Bone marrow mononuclear cells (BMMC) from 73 individuals (62 patients with either polycythemia vera (26), essential thrombocythemia (19), secondary polyglobuly (17) or chronic myeloid leukemia (2) and 11 healthy donors) were grown in parallel in the four media without, or with 0.01 U/ml erythropoietin (EPo). RESULTS: In all four media EEC formation was specific, as it was not observed in cultures of patients with secondary polyglobuly or chronic myeloid leukemia, nor of healthy donors. Analysis of fresh or MGG-stained collagen gel cultures allowed detection of EEC formation significantly more frequently than methylcellulose-based media; addition of 0.01 U/ml of EPo had little or no effect on EEC formation. Collagen based medium C1 gave better results than the other media tested: the 'C1' EEC assay was positive for 68.2% of polycythemia vera cultures with significantly higher median EEC numbers (6.5/10(5) BMMC for patients with one major criteria of polycythemia vera and 19 and 21/10(5) BMMC for patients with two or three major criteria, respectively). Medium C1 was also better for essential thrombocythemia cultures with 47.4% of positive results but with a low median EEC number (6.7/10(5) BMMC). When associated with the ELISA dosage of serum EPo, the 'C1' EEC assay allowed confirmation or elimination of the diagnosis of polycythemia vera for 91% (20/22) of polyglobulic patients. CONCLUSION: We propose that serum free collagen-based culture systems be considered to standardize the EEC assay, now part of the new criteria of polycythemia vera. PMID- 11920281 TI - Ovarian sarcoma and acute myelogenous leukaemia. PMID- 11920282 TI - A case of pancreatitis associated with all-trans-retinoic acid therapy in acute promyelocytic leukemia. PMID- 11920286 TI - Common deletion of SMAD4 in juvenile polyposis is a mutational hotspot. AB - Juvenile polyposis (JP) is an autosomal dominant syndrome in which affected patients develop upper- and/or lower-gastrointestinal (GI) polyps. A subset of families with JP have germline mutations in the SMAD4 (MADH4) gene and are at increased risk of GI cancers. To date, six families with JP have been described as having the same SMAD4 deletion (1244-1247delAGAC). The objective of the present study is to determine whether this deletion is a common ancestral mutation or a mutational hotspot. DNA from members of four families with JP, from Iowa, Mississippi, Texas, and Finland, that had this 4-bp deletion was used to genotype 15 simple tandem repeat polymorphism (STRP) markers flanking the SMAD4 gene, including 2 new STRPs within 6.3 and 70.9 kb of the deletion. Haplotypes cosegregating with JP in each family were constructed, and the distances of the closest markers were determined from the draft sequence of the human genome. No common haplotype was observed in these four families with JP. A 14-bp region containing the deletion had four direct repeats and one inverted repeat. Because no common ancestor was suggested by haplotype analysis and the sequence flanking the deletion contains repeats frequently associated with microdeletions, this common SMAD4 deletion in JP most likely represents a mutational hotspot. PMID- 11920285 TI - PARK3 influences age at onset in Parkinson disease: a genome scan in the GenePD study. AB - Parkinson disease (PD) is a late-onset neurodegenerative disorder. The mean age at onset is 61 years, but the disease can range from juvenile cases to cases in the 8th or 9th decade of life. The parkin gene on chromosome 6q and loci on chromosome 1p35-36 and 1p36 are responsible for some cases of autosomal recessive early-onset parkinsonism, but they do not appear to influence susceptibility or variability of age at onset for idiopathic PD. We have performed a genomewide linkage analysis using variance-component methodology to identify genes influencing age at onset of PD in a population of affected relatives (mainly affected sibling pairs) participating in the GenePD study. Four chromosomal loci showed suggestive evidence of linkage: chromosome 2p (maximum multipoint LOD [MaxLOD] = 2.08), chromosome 9q (MaxLOD = 2.00), chromosome 20 (MaxLOD = 1.82), and chromosome 21 (MaxLOD = 2.21). The 2p and 9q locations that we report here have previously been reported as loci influencing PD affection status. Association between PD age at onset and allele 174 of marker D2S1394, located on 2p13, was observed in the GenePD sample (P=.02). This 174 allele is common to the PD haplotype observed in two families that show linkage to PARK3 and have autosomal dominant PD, which suggests that this allele may be in linkage disequilibrium with a mutation influencing PD susceptibility or age at onset of PD. PMID- 11920288 TI - Incidence and risk factors for acute hepatitis B in the United States, 1982-1998: implications for vaccination programs. AB - From 1982-1998, enhanced sentinel surveillance for acute hepatitis B was conducted in 4 counties in the United States to determine trends in disease incidence and risk factors for infection. During this period, the reported incidence of acute hepatitis B declined by 76.1% from 13.8 cases per 100,000 in 1987 to 3.3 cases per 100,000 in 1998. Cases associated with injection drug use (IDU) decreased by 90.6%, men who have sex with men (MSM) by 63.5%, and heterosexual activity by 50.7%. During 1994-1998, the most commonly reported risk factor for infection was high-risk heterosexual activity (39.8%) followed by MSM activity (14.6%) and IDU (13.8%). Over half of all patients (55.5%) reported treatment for a sexually transmitted disease (STD) or incarceration in a prison or jail prior to their illness, suggesting that more than half of the acute hepatitis B cases might have been prevented through routine hepatitis B immunization in STD clinics and correctional health care programs. PMID- 11920287 TI - Mapping of gene loci for nephronophthisis type 4 and Senior-Loken syndrome, to chromosome 1p36. AB - For nephronophthisis (NPHP), the primary genetic cause of chronic renal failure in young adults, three loci have been mapped. To identify a new locus for NPHP, we here report on total-genome linkage analysis in seven families with NPHP, in whom we had excluded linkage to all three known NPHP loci. LOD scores >1 were obtained at nine loci, which were then fine mapped at 1-cM intervals. Extensive total-genome haplotype analysis revealed homozygosity in one family, in the region of the PCLN1 gene. Subsequent mutational analysis in this gene revealed PCLN1 mutations, thereby allowing exclusion of this family as a phenocopy. Multipoint linkage analysis for the remaining six families with NPHP together yielded a maximum LOD score (Z(max)) of 8.9 (at D1S253). We thus identified a new locus, NPHP4, for nephronophthisis. Markers D1S2660 and D1S2642 are flanking NPHP4 at a 2.9-cM critical interval. In one family with NPHP4, extensive genealogical studies were conducted, revealing consanguinity during the 17th century. On the basis of haplotype sharing by descent, we obtained a multipoint Z(max) of 5.8 for D1S253 in this kindred alone. In addition, we were able to localize to the NPHP4 locus a new locus for Senior-Loken syndrome, an NPHP variant associated with retinitis pigmentosa. PMID- 11920289 TI - CD4 T helper type 1 and regulatory T cells induced against the same epitopes on the core protein in hepatitis C virus-infected persons. AB - The factors that determine persistence or clearance of hepatitis C virus (HCV) infection are poorly understood. The CD4 T cell responses to the HCV core protein were examined in a cohort of women infected with a single genotype of HCV. CD4 T cells from HCV-infected patients secreted interferon (IFN)-gamma in response to peptides from 4 immunodominant regions of the core protein, and these responses were stronger in persistently infected women. Interleukin (IL)-10 was also produced by CD4 T cells from HCV-infected subjects in response to the same core peptides. Furthermore, HCV core-specific CD4 T cell clones secreted either IFN gamma or IL-10 but not IL-4. These findings demonstrate that T helper type 1 and regulatory T cells are induced against the same epitopes on the core protein during HCV infection. PMID- 11920290 TI - Influence of CD4 cell counts on the genetic heterogeneity of hepatitis C virus in patients coinfected with human immunodeficiency virus. AB - To study the effects of reduced CD4 T cell activity on hepatitis C virus (HCV) genetic heterogeneity, HCV quasi-species complexity and diversification over time were analyzed for 56 human immunodeficiency virus-coinfected patients. Patients were selected retrospectively from the French Seroconverter Cohort (SEROCO) and the French Hemophilia Cohort (HEMOCO) for having stable CD4 cell counts for 3 years. HCV complexity was assessed by single-strand conformation polymorphism analysis of the envelope-coding region (HVR) and the core region at 2 time points 3 years apart. Increased HVR complexity was associated with higher CD4 cell count and HCV genotype 1 infection. Qualitative variation of HVR and core region was not related to CD4 cell count and depended on the initial complexity. Complexity of both regions remained unchanged over 3 years. Among these HCV-HIV-coinfected patients with stable CD4 cell counts, viral genotype and CD4 cell count may have influenced HVR complexity before their inclusion in the study but were not involved in HVR diversification during the 3-year follow-up period. PMID- 11920291 TI - Similar frequency of rhinovirus-infectible cells in upper and lower airway epithelium. AB - Rhinovirus (RV) infections can alter lower airway physiology and inflammation, yet the characteristics of RV replication in lower airway cells are incompletely understood. An RV serotype 16 (RV16)-specific monoclonal antibody was identified. Immunohistochemistry and an infectious center assay were used to quantitate the infectivity of RV16 in primary bronchial and adenoidal epithelial cells. The proportion of infectible epithelial cells increased with the inoculum but did not exceed 10%. Analysis of bronchial tissue samples infected ex vivo demonstrated a small subset of RV-infected cells in the epithelial layer. These data confirm previous reports that RV infects only a small subset of epithelial cells in upper airway tissues and indicate that lower airway epithelial cells have a similar susceptibility to RV infection. In confirming that RV can infect cells in the lower airway, these results suggest that lower airway dysfunction occurs through this mechanism in susceptible persons. PMID- 11920292 TI - Molecular diagnosis of human enteroviruses by phylogeny-based classification by use of the VP4 sequence. AB - Human enteroviruses (EVs) are the major cause of a variety of acute and chronic illnesses. Virus isolation and neutralization tests are usually done to identify the causative virus, but these tests are labor intensive, time consuming, and sometimes require suckling mice from which certain viruses have been isolated. This study investigated a rapid and reliable method based on reverse transcription polymerase chain reaction and phylogenetic analysis. The phylogenetic tree constructed by neighbor-joining on the basis of the VP4 sequence from 66 prototypes grouped all human EVs into 5 distinct clusters. These clusters correspond closely to the 5 newly designated species-human EV A-D and poliovirus. The VP4 sequences of 89 isolates from 26 serotypes obtained over >30 years plus those of 66 prototype strains were analyzed. Each isolate formed a monophyletic cluster along with its respective prototype strain, allowing for serotype identification (with the exception of E-8). VP4-based classification appears to be an effective tool for the molecular epidemiology study of EVs. PMID- 11920293 TI - Human immunodeficiency virus (HIV)-specific effector CD8 T cell activity in patients with primary HIV infection. AB - The interferon-gamma ELISPOT assay was used to assess and compare the magnitude and breadth of human immunodeficiency virus (HIV)-specific CD8 T cell responses in treatment-naive subjects during the first year of HIV primary infection and during the chronic phase of infection. Twenty-five subjects tested within a year of exposure to HIV resulting in seroconversion and 20 subjects with chronic infection were screened for HIV peptide-specific activity by stimulating peripheral blood mononuclear cells with a panel of 5-21 HLA class I-restricted HIV peptides (mean, 11.2 +/- 3.5 HIV peptides). There was a significant correlation between the magnitude and breadth of HIV-specific effector responses and time elapsed from exposure (r=0.63 for magnitude vs. time and r=0.64 for breadth vs. time; P<.02, paired t test). Maximal breadth of the HIV gene product reactivity was achieved within 2 months of exposure for Nef-specific responses and by 4 months of exposure for responses directed to Env, Gag, and reverse transcriptase. PMID- 11920294 TI - The impact of persistent gastrointestinal colonization on the transmission dynamics of vancomycin-resistant enterococci. AB - The transmission dynamics of vancomycin-resistant enterococci (VRE) and factors contributing to their dissemination are complex. Mathematical modeling was used to simulate patterns of dissemination among patients and health care workers (HCWs) and to quantify the contribution of specific factors and infection control interventions on the endemic prevalence (EP) of VRE in a long-term hemodialysis unit. The model predicted that (1) an EP of 12% would be reached over time, regardless of the number of patients initially colonized, (2) endemicity would be sustained by the constant influx of newly colonized patients discharged from the hospital, (3) duration of VRE gastrointestinal colonization would have the most impact on the number of secondary cases, increasing the EP to a maximum of 70%, and (4) decreasing the patient:HCW ratio or improving hand hygiene would decrease the EP to 3%. Decreasing the duration of colonization, limiting hospital acquisition of VRE, and improving compliance with hand hygiene in the hemodialysis unit may decrease the rapidly rising rates of VRE in the patient population. PMID- 11920295 TI - Phylogenetic distribution of virulence-associated genes among Escherichia coli isolates associated with neonatal bacterial meningitis in the Netherlands. AB - Seventy cerebrospinal fluid Escherichia coli isolates from infants with neonatal bacterial meningitis (NBM), as submitted to the Netherlands Reference Laboratory for Bacterial Meningitis from 1989 through 1997, were assessed for phylogenetic background and extended virulence genotypes, in comparison with the E. coli reference collection, by using molecular methods. Phylogenetic group B2 significantly predominated overall (81%). The 4 major phylogenetic clusters exhibited distinctive virulence genotypes, suggesting diverse evolutionary histories for the individual genes. Many genes not previously studied in NBM, notably diarrhea-associated cdtB (cytolethal distending toxin [46%]) and urinary tract infection-associated ompT (outer membrane protease T [96%]), were as or more prevalent than traditional NBM-associated traits, such as ibeA (invasion of brain endothelium [33%]), sfaS (S fimbriae [59%]), and K1 capsule (81%). These findings provide novel insights into the phylogenetic origins of NBM-associated E. coli and suggest numerous new potential targets for preventive interventions against this dire disease. PMID- 11920296 TI - Inhibition of shiga toxin cytotoxicity in human renal cortical epithelial cells by nitrobenzylthioinosine. AB - Nitrobenzylthioinosine (NBTI), a nucleoside-transport inhibitor, has been found to possess the ability to prevent the cytotoxic action of Shiga toxin (Stx) 1 in human renal cortical epithelial cells (HRCECs), thereby protecting HRCECs from cell death. Further examination revealed that NBTI does not affect either the binding or the endocytosis of Stx1 but alters the intracellular transport of Stx1. Generally, endocytosed Stx1 is thought to be transported from endosomes to the endoplasmic reticulum. In NBTI-treated cells, however, the endocytosed Stx1 is delivered to an early endosome, but no further transportation occurs. Moreover, Stx1 is rapidly excreted from NBTI-treated HRCECs, preventing the accumulation of Stx1. Investigation of the NBTI-mediated protection mechanism against Stx cytotoxicity may provide insights into the analysis of Stx-mediated cell damage and lead to improvements in therapeutic approaches for diseases caused by Stx. PMID- 11920298 TI - Bacterial endosymbionts of Onchocerca volvulus in the pathogenesis of posttreatment reactions. AB - Treatment of onchocerciasis with diethylcarbamazine (DEC) or ivermectin is associated with a posttreatment reaction characterized by fever, tachycardia, hypotension, lymphadenopathy, and pruritus. To investigate the role of the Wolbachia bacterial endosymbiont of Onchocerca volvulus in these reactions, serum samples collected before and after treatment with either anthelmintic were assessed for evidence of Wolbachia DNA. By use of real-time quantitative polymerase chain reaction, Wolbachia DNA was detected in both groups-with significantly higher levels in those who received DEC (P <.0001). In the ivermectin group, there was a significant correlation between levels of bacterial DNA and serum tumor necrosis factor-alpha (P =.013). Peak DNA levels correlated with reaction scores (P =.048). Significant correlations were also seen between Wolbachia DNA and the antibacterial peptides calprotectin (P =.021) and calgranulin B (P <.0001). These findings support a role for Wolbachia products in mediating the inflammatory responses seen following treatment of onchocerciasis and suggest new targets for modulating these reactions. PMID- 11920297 TI - Borrelia burgdorferi are susceptible to killing by a variety of human polymorphonuclear leukocyte components. AB - The killing of Borrelia burgdorferi by intact human polymorphonuclear leukocytes (PMNL) and by individual PMNL components was compared. Intact PMNL killed B. burgdorferi 6.5-fold more efficiently and 5-fold more completely when spirochetes were opsonized with specific antibodies. U-cytoplasts, which have activatable oxidase, killed opsonized B. burgdorferi with an efficiency similar to that of intact PMNL in killing unopsonized B. burgdorferi. Although B. burgdorferi were susceptible to H(2)O(2) and nitric oxide, PMNL lysates killed B. burgdorferi nearly as well as intact PMNL killed opsonized B. burgdorferi, suggesting a critical role for granule contents. B. burgdorferi were killed by the PMNL antimicrobial components elastase, LL-37, bactericidal/permeability-increasing protein, and human neutrophil peptide-1. B. burgdorferi had limited susceptibility to killing by lysozyme and were not killed by azurocidin, proteinase 3, or lactoferrin. The efficient killing of B. burgdorferi by a variety of PMNL mechanisms highlights the paradoxical persistence of spirochetes in vivo. PMID- 11920299 TI - CD4 T cell responses to a variant antigen of the malaria parasite Plasmodium falciparum, erythrocyte membrane protein-1, in individuals living in malaria endemic areas. AB - Plasmodium falciparum erythrocyte membrane protein-1 (PfEMP-1) is a variant antigen on the surface of malaria-infected red blood cells. Antibody responses to PfEMP-1 correlate with immunity, and, therefore, PfEMP-1 may be a good candidate for a malaria vaccine. However, the specificity of CD4 T cells required for a protective variant-specific antibody response is not known. We have measured the CD4 T cell response to 3 different regions that are relatively homologous among different PfEMP-1 variants. The response to the cysteine-rich interdomain region was unusual in that the majority of donors, whether malaria exposed or not, had positive CD4 T cell, interleukin-10, and interferon-gamma responses. The CD4 T cell response to the exon 2 and duffy binding-like domain proteins was significantly greater in malaria-exposed donors than in unexposed Europeans, which suggests that these regions contain peptides recognized by T cells, which thus may be useful as components of a vaccine. PMID- 11920301 TI - Cellular immunity in monozygotic twins discordant for chronic fatigue syndrome. AB - Studies elsewhere have suggested that immune dysfunction may be common in patients with chronic fatigue syndrome (CFS). The objective of this study was to assess the nature and extent of abnormalities in lymphocyte cell surface markers and NK cell activity in patients with CFS while controlling for genetic factors. A co-twin control study of immune system parameters was conducted for 22 pairs of monozygotic twins discordant for CFS and 9 healthy pairs of twins. The CFS twins had greater numbers of CD62L(+) T cells in several T cell subsets, although these differences did not achieve statistical significance. Significantly greater variability was noted in twins discordant for CFS than in the concordant healthy twins for 20 of 48 variables examined. The monozygotic co-twin control design is of unique value because of its ability to control for genetic influences on CFS; however, additional studies will be required to further assess immune dysregulation in this illness. PMID- 11920300 TI - A recombinant blood-stage malaria vaccine reduces Plasmodium falciparum density and exerts selective pressure on parasite populations in a phase 1-2b trial in Papua New Guinea. AB - The malaria vaccine Combination B comprises recombinant Plasmodium falciparum ring-infected erythrocyte surface antigen and 2 merozoite surface proteins (MSP1 and MSP2) formulated in oil-based adjuvant. A phase 1-2b double-blind, randomized, placebo-controlled trial in 120 children (5-9 years old) in Papua New Guinea demonstrated a 62% (95% confidence limits: 13%, 84%) reduction in parasite density in children not pretreated with sulfadoxine-pyrimethamine. Vaccinees had a lower prevalence of parasites carrying the MSP2-3D7 allelic form (corresponding to that in the vaccine) and a higher incidence of morbid episodes associated with FC27-type parasites. These results demonstrate functional activity of Combination B against P. falciparum in individuals with previous malaria exposure. The specific effects on parasites with particular msp2 genotypes suggest that the MSP2 component, at least in part, accounted for the activity. The vaccine-induced selection pressure exerted on the parasites and its consequences for morbidity strongly argue for developing vaccines comprising conserved antigens and/or multiple components covering all important allelic types. PMID- 11920302 TI - Human papillomavirus DNA in oral mucosal lesions. AB - This study determined the presence of human papillomavirus (HPV) DNA in oral mucosa cells from 121 patients with different types of oral mucosal lesions (13 squamous cell carcinomas, 59 potentially malignant lesions, 49 benign erosive ulcerative lesions) and from 90 control subjects. HPV DNA was detected by nested polymerase chain reaction, and genotype was determined by DNA sequencing. HPV prevalence was 61.5% in carcinomas, 27.1% in potentially malignant lesions, 26.5% in erosive ulcerative lesions, and 5.5% in control subjects. The risk of malignant or potentially malignant lesions was associated with HPV and was statistically significant. HPV-18 was found in 86.5% of HPV-positive lesions but was not associated with a particular type of lesion and was found in 80% of the HPV-positive control subjects. HPV infection was related to older age but not to sex, smoking, or alcohol use; the presence of lesions in the oral cavity increased the risk of HPV infection. PMID- 11920303 TI - Ciprofloxacin resistance in Campylobacter jejuni evolves rapidly in chickens treated with fluoroquinolones. AB - Fluoroquinolones are commonly used to treat gastroenteritis caused by Campylobacter species. Domestically acquired fluoroquinolone-resistant Campylobacter infection has been documented recently in the United States. It has been proposed that the increase in resistance is due, in part, to the use of fluoroquinolones in poultry. In separate experiments, the effects of sarafloxacin and enrofloxacin treatment of Campylobacter jejuni-infected chickens on the development of ciprofloxacin resistance were measured. Fecal samples were collected before and after treatment and were cultured for C. jejuni. When enrofloxacin or sarafloxacin was used at US Food and Drug Administration-approved doses in broiler chickens, resistance developed rapidly and persisted in C. jejuni. MICs of ciprofloxacin increased from a base of 0.25 microg/mL to 32 microg/mL within the 5-day treatment time frame. These results show that the use of these drugs in chickens rapidly selects for resistant Campylobacter organisms and may result in less effective fluoroquinolone therapy for cases of human campylobacteriosis acquired from exposure to contaminated chicken. PMID- 11920304 TI - A pseudo-outbreak of Chlamydia trachomatis in a state residential facility: implications for diagnostic testing. AB - In December 1998, an outbreak of Chlamydia trachomatis genital infections was reported among 18 residents of a state residential facility housing 392 mentally retarded clients. The initial patient tested positive by ligase chain reaction (LCR); 17 others tested positive by culture. Serologic test results for C. trachomatis antibodies in patients who had tested positive by culture were negative. Further testing showed that C. trachomatis DNA could not be detected in the LCR specimen or in any reportedly positive culture specimens. At the original culture laboratory, C. trachomatis culture was infrequently performed, and positive controls were not adequately prepared. This pseudo-outbreak highlights problems that may occur with C. trachomatis testing. As experience with C. trachomatis culture declines, laboratories performing this test should ensure quality and consider confirmatory testing. For C. trachomatis screening tests, the need for confirmatory testing depends on individual patient considerations (including medical-legal implications) and prevalence of infection in the tested population. PMID- 11920305 TI - Field effectiveness of Vi polysaccharide typhoid vaccine in the People's Republic of China. PMID- 11920307 TI - Human herpesvirus 6 viremia in bone marrow transplant recipients: clinical features and risk factors. AB - Human herpesvirus 6 (HHV-6) infection was studied in 82 bone marrow transplant (BMT) recipients (72 allogeneic, 10 autologous). All recipients and 30 donors were seropositive for HHV-6 antibody at the time of bone marrow transplantation. Thirty-one recipients (37.8%) had HHV-6 viremia 2-4 weeks after transplantation. The incidence of HHV-6 viremia was significantly higher among allogeneic BMT recipients than in autologous BMT recipients (P=.011). Therefore, the following analyses of allogeneic BMT recipients were carried out (n=72). Geometric mean antibody titers (log(10)) were significantly higher in recipients without viremia than in those with viremia (1.84+/-0.39 vs. 1.61+/-0.42; P=.022). Logistic regression analysis demonstrated that leukemia or lymphoma is an independent risk factor (P=.031) for HHV-6 viremia. Rash occurring within 1 month after transplantation was observed in 17 (54.8%) of 31 recipients with HHV-6 viremia but in only 8 (19.5%) of 41 recipients without HHV-6 viremia (P=.001). PMID- 11920308 TI - Preemptive use of oral ganciclovir to prevent cytomegalovirus infection in liver transplant patients: a randomized, placebo-controlled trial. AB - The use of postdetection antiviral treatment of cytomegalovirus (CMV) as a strategy to prevent infection and disease in solid-organ transplant patients has not been evaluated by placebo-controlled trials. We carried out such a study in 69 patients who had received liver transplants and had positive results of CMV polymerase chain reaction within 8 weeks after transplantation but did not have concomitant CMV infection or disease. These patients were randomly assigned to receive placebo or oral ganciclovir for 8 weeks. CMV infection developed in 21% and disease developed in 12% of placebo recipients (P =.022), compared with 3% and 0%, respectively, among ganciclovir recipients (P =.003). Similarly, in the placebo arm, 55% and 36% of CMV-negative patients who received organs from CMV positive donors developed CMV infection or disease, respectively (P =.02), compared with 11% and 0% of such patients in the ganciclovir arm (P <.01). Oral ganciclovir administered on CMV detection by PCR prevents CMV infection or disease after liver transplantation. PMID- 11920309 TI - Comparison of cytomegalovirus (CMV) UL97 gene sequences in the blood and vitreous of patients with acquired immunodeficiency syndrome and CMV retinitis. AB - Cytomegalovirus (CMV) resistance to ganciclovir occurs via mutations in the UL97 gene. CMV DNA, from vitreous and blood specimens and from culture isolates from 87 patients with acquired immunodeficiency syndrome and CMV retinitis who received a ganciclovir implant, was sequenced to identify the relationship between the UL97 DNA sequences in the eye and peripheral blood. There was 93.5% agreement between the UL97 gene sequences from paired vitreous specimens and blood specimens. Sequence analysis of vitreous specimens showed that 15% (13/87) of the patients had either a ganciclovir resistance-conferring mutation or a polymorphism in the CMV UL97 gene. Eleven of the 13 mutations or polymorphisms in the vitreous also were identified in blood. Although the number of mutations limits definitive interpretation, these data suggest that blood specimens may reflect the events occurring in the eyes of patients with CMV retinitis. PMID- 11920310 TI - Relationship of hepatitis C genotype 1 NS5A sequence mutations to early phase viral kinetics and interferon effectiveness. AB - The interferon (IFN) sensitivity-determining region (ISDR) of the hepatitis C virus (HCV) NS5A protein is controversially implicated in determining IFN-alpha sustained viral response for HCV genotype 1-infected patients. Because the NS5A protein interferes with protein kinase antiviral activity, this study attempted to determine whether ISDR amino acid mutation number would correlate better with IFN effectiveness to inhibit virus production and elicit early virus clearance. Early viral kinetic data from 22 genotype 1-infected patients treated with high dose IFN was compared with ISDR mutation number. IFN effectiveness and first phase viral log decline correlated directly with ISDR mutation number (P=.02). Mean mutation number was higher among rapid responders (3.7+/-1.0; n=6) than among nonresponders (1.3+/-1.0; n=16) (P=.001). Also, second-phase viral log decline and calculated hepatocyte death rate correlated directly with ISDR mutation number (P=.01). This supports a dual effector role for NS5A, which regulates virus production and immune clearance early in therapy. PMID- 11920311 TI - Parenteral vaccination against influenza does not induce a local antigen-specific immune response in the nasal mucosa. AB - The immune response in the nasal mucosa to influenza vaccination in 23 patients scheduled for tonsillectomy was studied. A statistically significant increase in influenza virus-specific serum and oral fluid antibodies was observed 7 days after vaccination. The numbers of influenza virus-specific antibody-secreting cells (ASCs) in peripheral blood also increased significantly 1 week after vaccination. The numbers of ASCs in tonsils and nasal mucosa were compared with data from a recent study of nonvaccinated volunteers. The numbers of influenza virus-specific ASCs in tonsils were significantly higher in the vaccinated group, but, surprisingly, there was no significant difference between the groups in the numbers of ASCs in nasal mucosa. This suggests that the influenza virus-specific antibodies detected in oral fluid are not produced locally in the nasal mucosa and may originate from a systemic source, indicating that the vaccination may favor a systemic immune response. PMID- 11920312 TI - Regulation of CC chemokine receptor 5 and CD4 expression and human immunodeficiency virus type 1 replication in human macrophages and microglia by T helper type 2 cytokines. AB - Macrophages, microglia, and other mononuclear phagocytes serve as cellular reservoirs for viral persistence in patients with acquired immunodeficiency syndrome. To understand host mechanisms that affect human immunodeficiency virus type 1 (HIV-1) pathogenesis by modulating expression of coreceptors, cytokine regulation of CC chemokine receptor 5 (CCR5) and CD4 expression on monocytes, monocyte-derived macrophages (MDMs), and microglia was investigated. Interleukin (IL)-4 and IL-10 enhanced the entry and replication of HIV-1 in microglia through up-regulation of CD4 and CCR5 expression, respectively. IL-4 stimulated HIV-1 replication in MDMs but down-regulated CD4 and CCR5 expression and inhibited virus entry, whereas IL-10 had the opposite effects. Thus, mechanisms independent of CCR5 and CD4 expression levels are involved in pathways that regulate HIV-1 replication in MDMs. CCR5 up-regulation by IL-10 was associated with increased migration of microglia in response to macrophage inflammatory protein-1beta. These findings suggest that increased production of T helper type 2 cytokines in the later stages of disease can enhance virus entry and replication in mononuclear phagocytes and facilitate chemotactic migration. PMID- 11920313 TI - Broad nucleoside-analogue resistance implications for human immunodeficiency virus type 1 reverse-transcriptase mutations at codons 44 and 118. AB - Two large, independent human immunodeficiency virus type 1 resistance databases containing >7700 reverse-transcriptase (RT) sequences were used to analyze the epidemiology of amino acid substitutions at codons 44 and 118, which confer moderate lamivudine resistance in the presence of zidovudine resistance. As expected, E44A/D and V118I mutations were strongly associated with M41L, D67N, L210W, and T215Y but also with other mutations, including K43E/N/Q, T69D, V75M, H208Y, R211K, and K219R. Both E44D and V118I were more frequently associated with stavudine and didanosine than with zidovudine and lamivudine treatment. However, selection of E44A/D and V118I was also detected in association with a switch to other nucleoside RT inhibitors, including zalcitabine and abacavir. Site-directed mutagenesis confirmed that 44D and 118I can decrease phenotypic susceptibility not only to lamivudine but also to most other nucleoside analogues, particularly stavudine and abacavir. Thus, substitutions at RT codons 44 and 118 have broad implications in nucleoside RT inhibitor resistance in the setting of several nucleoside-associated mutations. PMID- 11920314 TI - Equal plasma viral loads predict a similar rate of CD4+ T cell decline in human immunodeficiency virus (HIV) type 1- and HIV-2-infected individuals from Senegal, West Africa. AB - Human immunodeficiency virus (HIV) type 2 infection is characterized by slower disease progression to acquired immunodeficiency syndrome than results from HIV-1 infection. To better understand the biological factors underlying the different natural histories of infection with these 2 retroviruses, we examined the relationship between HIV RNA and DNA levels and the rate of CD4(+) T cell decline among 472 HIV-1- and 114 HIV-2-infected individuals from Senegal. The annual rate of CD4(+) T cell decline in the HIV-2 cohort was approximately one-fourth that seen in the HIV-1 cohort. However, when the analysis was adjusted for baseline plasma HIV RNA level, the rates of CD4(+) T cell decline per year for the HIV-1 and HIV-2 cohorts were similar (a rate increase of approximately 4% per year for each increase in viral load of 1 log(10) copies/mL). Therefore, plasma HIV load is predictive of the rate of CD4(+) T cell decline over time, and the correlation between viral load and the rate of decline appears to be similar among all HIV infected individuals, regardless of whether they harbor HIV-1 or HIV-2. PMID- 11920315 TI - The heterogeneity of endemic community pediatric group a streptococcal pharyngeal isolates and their relationship to invasive isolates. AB - By use of molecular techniques, the genetic heterogeneity of 63 community pediatric pharyngeal group A streptococcal (GAS) isolates circulating within a 3 week period were compared with 17 contemporaneous invasive pediatric isolates. Pharyngitis isolates represented 16 pulsed-field gel electrophoresis (PFGE) patterns with 12 emm serotypes, and invasive isolates represented 10 PFGE patterns with 9 emm serotypes. One-fourth of the pharyngeal isolates (16/63) were identical to at least 1 invasive isolate; conversely, 10 (59%) of 17 invasive isolates were identical to at least 1 pharyngeal strain. sic allele analysis of emm1 strains demonstrated additional heterogeneity and overlap. More pharyngeal (71%) than invasive isolates (35%) were positive for both speA and speC (P<.02). Many pharyngitis GAS strains circulate simultaneously. Most invasive pediatric GAS strains are identical to acute pharyngitis strains; thus, childhood pharyngitis is a major reservoir for strains with invasive potential. Pharyngeal isolates were more likely to be speA and speC positive than were the invasive isolates. PMID- 11920316 TI - Differential effects of p38- and extracellular signal-regulated kinase mitogen activated protein kinase inhibitors on inducible nitric oxide synthase and tumor necrosis factor production in murine macrophages stimulated with Streptococcus pneumoniae. AB - The role of p38- and extracellular signal-regulated kinase (ERK) mitogen activated protein (MAP) kinase pathways in the up-regulation of inducible nitric oxide synthase (iNOS) and tumor necrosis factor (TNF) production in macrophages stimulated with Streptococcus pneumoniae was examined. Inhibitors of p38 kinases effected significant decreases in the accumulation of iNOS protein in macrophages challenged with pneumococcal cell wall preparations or antibiotic-killed pneumococci, even when added up to 6 h after bacterial challenge. In contrast, ERK pathway inhibitors failed to inhibit pneumococcus-induced iNOS protein accumulation. ERK pathway inhibitors significantly reduced TNF secretion when added at the same time as pneumococcal challenge, and inhibitors of both ERK and p38 pathways reduced TNF secretion when added to the macrophages 1 h before stimulation. These data confirm the importance of the p38 and ERK MAP kinase pathways in macrophage activation by bacterial products but indicate that these 2 kinase pathways regulate different macrophage responses in a temporally distinct manner. PMID- 11920317 TI - Reduction of nasopharyngeal carriage of Streptococcus pneumoniae after administration of a 9-valent pneumococcal conjugate vaccine to toddlers attending day care centers. AB - A double-blind, randomized study involving 264 toddlers attending day care centers was conducted to document the effect of a 9-valent pneumococcal conjugate vaccine on the carriage rate of pneumococci. Of 3750 cultures done on nasopharyngeal samples obtained from subjects during a 2-year follow-up period after vaccination, 65% were positive for Streptococcus pneumoniae. In all age windows, the rate of carriage of vaccine-type pneumococci was lower among subjects who received the pneumococcal vaccine than among control subjects, because the acquisition rate was lower in the former group. The effect was most pronounced among subjects aged < or =36 months. The sample size enabled us to study protection against carriage of S. pneumoniae serotypes 6B, 9V, 14, 19F, and 23F; significant protection against all serotypes except 19F was seen in the pneumococcal-vaccine group. The rate of carriage of serotype 6A (not included in the vaccine) was also reduced significantly, but the rate of carriage of serotype 19A (not included in the vaccine) was not. The rate of carriage of non-vaccine type pneumococci (excluding serotype 6A) was higher in the pneumococcal-vaccine group than in the control group. PMID- 11920318 TI - Inhibition of staphylococcal wound infection and potentiation of antibiotic prophylaxis by a recombinant fragment of the fibronectin-binding protein of Staphylococcus aureus. AB - Adherence of Staphylococcus aureus to host tissues is a critical step for colonization and initiation of infection. The fibronectin-binding proteins (FnBPs) of S. aureus have been implicated in adherence and internalization in nonprofessional phagocytes. A recombinant fragment of the fibronectin-binding domains (rFnBF) that potently inhibits S. aureus entry into host cells was generated. To test the hypothesis that rFnBF may attenuate the establishment of infection, the ability of intermuscularly administered rFnBF to prevent abscess formation was determined in a guinea pig model of wound infection. rFnBF exhibited dose-dependent inhibition of abscess formation and, at a 100-microg dose, raised the median infective dose approximately 170-fold, compared with the control. In addition, rFnBF potentiated the benefit of prophylaxis with cefazolin. Thus, exogenous administration of the fibronectin-binding domain of FnBP reduces the risk of staphylococcal abscess formation and should be investigated further as a novel agent for prevention of wound infection. PMID- 11920319 TI - Markers of inflammation in bacterial diarrhea among travelers, with a focus on enteroaggregative Escherichia coli pathogenicity. AB - The intestinal inflammatory response of traveler's diarrhea acquired in Goa, India, and Guadalajara, Mexico, was studied. Fecal lactoferrin was found in stool samples in which enteroaggregative Escherichia coli (EAEC), enterotoxigenic E. coli, or Salmonella or Shigella species were isolated, with Shigella-positive cases showing the highest level. Samples from cases of Shigella-associated diarrhea had the highest concentrations of fecal cytokines. Travelers to India who had EAEC-associated diarrhea showed elevated levels of interleukin (IL)-8 (median, 341.15 pg/mL) and IL-1beta (median, 749.90 pg/mL). Although 15 travelers to Mexico who had EAEC-associated diarrhea had a median concentration of 0 pg/mL for both IL-8 and IL-1beta, 2 had high levels of IL-8 (1853 and 11,786 pg/mL), and 5 showed elevated levels of IL-1beta (1-1240 pg/mL). Samples from patients in India who had pathogen-negative diarrhea or from patients in Mexico who had asymptomatic EAEC infection were negative for cytokines. Bacterial pathogens causing traveler's diarrhea commonly produce intestinal inflammation, although a subset of patients with EAEC-associated diarrhea fail to develop an inflammatory response. PMID- 11920320 TI - Induction of protective immunity by synthetic Vibrio cholerae hexasaccharide derived from V. cholerae O1 Ogawa lipopolysaccharide bound to a protein carrier. AB - Synthetic antigens that mimic the terminal hexasaccharide epitope of the O specific polysaccharide of Vibrio cholerae O1, serotype Ogawa, were conjugated to bovine serum albumin (BSA). Conjugates with carbohydrate-to-carrier molar ratios of 15.5:1, 9.2:1, and 4.6:1 were tested for immunogenicity and efficacy in mice. The role of preimmunity to BSA and the use of adjuvant in the generation of the serologic response to the O-specific polysaccharide and protection against virulent V. cholerae was examined. Preimmunity to BSA did not affect the anti Ogawa titers but seemed to enhance the protective capacity of antiserum. All 3 conjugates were immunogenic, but adjuvant was effective at inducing higher and earlier antibody responses. In tertiary serum samples, a correlation was found between vibriocidal activity and protection. The protective capacity of antiserum was evident in serum from mice immunized with all conjugates, but it was highest in the groups that received the conjugate with the lowest level of substitution. Further studies are required to increase understanding of the reason for differential protection. PMID- 11920321 TI - The role of endogenous interleukin (IL)-18, IL-12, IL-1beta, and tumor necrosis factor-alpha in the production of interferon-gamma induced by Candida albicans in human whole-blood cultures. AB - Despite the importance of interferon (IFN)-gamma, tumor necrosis factor (TNF), and interleukin (IL)-18 for host defense against candidiasis, the pathways leading to their stimulation by Candida albicans are unclear. In a whole-blood model, IL-18 neutralization by IL-18 binding protein decreased C. albicans induced IFN-gamma synthesis by 72%. Similarly, neutralization of IL-12 or IL 1beta by either neutralizing antibodies or IL-1 receptor antagonist also reduced (by 65%) IFN-gamma production. Neutralization of TNF by TNF binding proteins resulted in only a 36% reduction of IFN-gamma synthesis. In contrast, production of TNF and IL-8 was largely unaffected by these cytokine inhibitors. Thus, C. albicans stimulates IFN-gamma production in an IL-18-, IL-12-, and IL-1beta dependent manner, whereas production of TNF and IL-8 is independent of these cytokines. Blocking the biologic activities of IL-18, IL-12, and IL-1beta in patients (e.g., for treatment of autoimmune diseases) may result in increased susceptibility to C. albicans infection. PMID- 11920322 TI - Absolute levels and ratios of proinflammatory and anti-inflammatory cytokine production in vitro predict clinical immunity to Plasmodium falciparum malaria. AB - The relationship between malaria-related outcomes and cytokine production in whole blood cultures associated with cellular immune responses and immunity to Plasmodium falciparum malaria was examined in a study in southern Ghana. Production of malaria-specific interferon (IFN)-gamma was associated with reduced risk of fever and clinical malaria. Protective IFN-gamma responses were induced by live schizonts but not by dead parasites. Production of malaria-specific tumor necrosis factor (TNF)-alpha was associated with reduced risk of fever during follow-up. Baseline levels of TNF-alpha and phytohemagglutinin (PHA)-induced interleukin (IL)-10 were positively associated with hemoglobin concentration. IL 12 production was associated with reduced risk of parasitemia. PHA-induced transforming growth factor-beta production was associated with reduced risk of fever during follow-up. High ratios of proinflammatory to anti-inflammatory cytokines were associated with increased risk of fever and higher hemoglobin concentrations. Thus, absolute levels and ratios of proinflammatory and anti inflammatory cytokines influence susceptibility to infection, clinical disease, and anemia. These data contradict data from cross-sectional clinical studies and indicate a need for detailed analysis of the relationship between cellular immunity to malaria and resistance to disease. PMID- 11920323 TI - Molecular epidemiology of influenza A(H3N2) virus reinfections. AB - Between 1979 and 1989, families enrolled in the Houston Family Study were prospectively monitored for influenza virus infections. Reinfection with the H3N2 subtype occurred in a number of family members, and 6 pairs of isolates (interval between collection of first and second isolate, 2-5 years) were available for molecular analysis. Changes in the hemagglutinin genes of pairs of viruses isolated from the same individuals were examined to determine the molecular basis for reinfection. The findings of this study indicate that reinfection of an individual by viruses of the same subtype may occur within a relatively short period of time when the paired strains have genetically distinct hemagglutinin genes in which amino acid changes are present in the defined antigenic sites. PMID- 11920324 TI - Expression of human immunodeficiency virus coreceptors CXC chemokine receptor 4 and CC chemokine receptor 5 on monocytes is down-regulated during human endotoxemia. AB - Lipopolysaccharide (LPS) can inhibit human immunodeficiency virus (HIV) infection in monocytes in vitro. To test the hypothesis that an LPS effect on CXC chemokine receptor 4 (CXCR4) and CC chemokine receptor 5 (CCR5), known coreceptors for HIV, contributes to this effect, 8 healthy men were intravenously injected with Escherichia coli LPS (4 ng/kg), and monocyte CXCR4 and CCR5 expression was monitored by fluorescence-activated cell sorter analysis. LPS induced a decrease in the fraction of peripheral blood monocytes expressing CXCR4 and CCR5, reaching a nadir after 2 h (both P<.001 vs. baseline). In whole blood in vitro, not only LPS but also lipoarabinomannan (a cell wall component of Mycobacterium tuberculosis) and lipoteichoic acid (a cell wall component of Staphylococcus aureus) down-regulated the expression of CXCR4 and CCR5 on monocytes (all P<.05). Exposure of monocytes to (myco)bacterial agents may render them relatively resistant to infection with HIV by an effect on HIV coreceptors. PMID- 11920325 TI - Correlates of prevalent and incident Kaposi's sarcoma-associated herpesvirus infection in men who have sex with men. AB - Infection with Kaposi's sarcoma-associated herpesvirus (KSHV) is common among men who have sex with men (MSM). To determine correlates of infection, 578 human immunodeficiency virus (HIV)-negative MSM were assessed by serologic assays, questionnaires, and physical examinations. At baseline, 76 (16%) of 474 participants were KSHV seropositive. Prevalent KSHV infection was significantly associated with hepatitis A (odds ratio [OR], 3.3; 95% confidence interval [CI], 1.5-7.5), hepatitis B seropositivity (OR, 2.6; 95% CI, 1.4-4.8), herpes simplex virus (HSV)-2 (OR, 2.4; 95% CI, 1.3-4.4), and >4 male partners in the previous 6 months (OR, 1.9; 95% CI, 1.1-3.2). Fifteen KSHV seroconversions (4%) were observed for an incidence of 3.8/100 person-years, similar to HSV-1 incidence in this cohort and more frequent than incidence of HIV and HSV-2. Reporting > or =1 HIV-positive partner (OR, 5.9; 95% CI, 1.8-19.3), amyl nitrite use (OR, 7.0; 95% CI, 2.1-23.0), and lymphadenopathy in the past 6 months (OR, 7.7; 95% CI, 1.9 31.0) correlated with KSHV seroconversion. PMID- 11920326 TI - Formation of vegetations during infective endocarditis excludes binding of bacterial-specific host antibodies to Enterococcus faecalis. AB - Infectious endocarditis is a microbial infection of the endothelial lining of the heart that typically occurs on damaged or prosthetic heart valves. The characteristic lesion seen with infective endocarditis, termed "the vegetation," is composed of bacteria surrounded by a platelet/fibrin layer attached to the underlying endothelium. The vegetation has long been believed to exclude or hinder host defenses from clearing bacteria, although formal demonstration of mechanisms by which this occurs are lacking. This study investigated the ability of the vegetation to exclude host antibodies specific for the bacterial surface protein aggregation substance in vivo during experimental endocarditis caused by Enterococcus faecalis. The results demonstrate that, once the vegetation encloses bacteria, they are no longer accessible to high-titer bacterial-specific host antibodies, establishing a mechanism by which the vegetation functions to protect the bacteria from the humoral immune response. PMID- 11920327 TI - Role of interleukin-18 in human immunodeficiency virus type 1 infection. PMID- 11920329 TI - Effect of productive in vitro human immunodeficiency virus or simian immunodeficiency virus infection on telomerase activity in lymphoid and nonlymphoid cells. PMID- 11920331 TI - Ultrasonography in renal transplantation. AB - Sonography is a simple, inexpensive, and readily available imaging modality that has become an essential component of the management of renal transplantation. It is indicated in almost all patients with acute renal failure and also is useful in the evaluation of pain, infection, and hematuria and the performance of percutaneous biopsy. Although many aspects of sonography are similar in native and transplanted kidneys, there are important differences and problems unique to the renal allograft, which form the basis for this review. The anatomy of renal transplantation and changes that accompany parenchymal disorders are discussed, but particular attention focuses on problems related to the urinary tract, fluid collections, and vascular disorders. By becoming more familiar with transplant sonography, nephrologists will be better able to incorporate this indispensable tool into the care of their patients. PMID- 11920332 TI - Glomerular size and glomerulosclerosis: relationships to disease categories, glomerular solidification, and ischemic obsolescence. AB - In focal segmental glomerulosclerosis (FSGS), a biphasic change in glomerular size is described in which glomeruli are enlarged with early glomerulosclerosis. Hyperperfusion of larger glomeruli is believed to contribute to progressive glomerular injury. This study was undertaken to investigate whether similar alterations in glomerular size can be found in other renal diseases. Volumes of sclerotic and nonsclerotic glomeruli were estimated in renal biopsy specimens using the Weibel and Gomez method (1962). Glomerulosclerosis was graded on individual glomeruli from 0 to 4, with 0 as no sclerosis and 4 as 76% to 100% sclerosis. Primary and secondary FSGS showed a biphasic change in which grade 2 glomeruli were 50% larger than grade 0 glomeruli and grades 3 and 4 glomeruli were solidified and smaller than grade 0 glomeruli. In essential hypertension, no increase in glomerular size was seen with early glomerulosclerosis, and the latter stages consisted of ischemic obsolescence in which collapsed tufts were 50% smaller than solidified glomeruli of FSGS. Grade 0 glomeruli of membranous glomerulonephritis (MGN) were significantly larger than grade 0 glomeruli of FSGS, and no significant difference in size was seen between grades 0 and 2 glomerulosclerosis. Solidified diabetic glomeruli maintained a large size with grades 3 and 4 sclerosis. Glomeruloscleroses of FSGS, hypertension, MGN, and diabetes have stereologically distinct features. A biphasic change in glomerular size is characteristic of primary and secondary FSGS, but not hypertension, in which tuft collapse supports reduced rather than increased perfusion in the pathogenesis of its glomerular obsolescence. PMID- 11920333 TI - Congenital renal agenesis: case-control analysis of birth characteristics. AB - Renal agenesis (RA) is the most common congenital urinary malformation; however, evidence for prenatal factors associated with the development of this condition is limited. The aim of this study is to determine prenatal and perinatal factors associated with the development of RA. We performed a case-control study of Colorado births with RA from 1989 to 1998 identified in the Colorado Responds for Children With Special Needs and linked with birth certificate data. There were 189 infants with RA and 940 control infants without RA on the study. The incidence of RA is approximately 1 per 2,900 births. Seventy-six infants (40.6%) born with RA were girls, and 111 infants (59.4%) were boys (P = 0.02). Preexisting diabetes mellitus in the mother was a significant factor associated with RA (adjusted odds ratio (OR), 4.98; 95% confidence interval (CI), 1.08 to 22.93) while controlling for maternal age, maternal race, and alcohol exposure. Black maternal race also was significantly associated with the development of RA (adjusted OR, 2.19; 95% CI, 1.26 to 3.95) while controlling for other factors. Maternal age younger than 18 years, maternal education less than 12 years, alcohol exposure, and infant sex approached significance. RA is a relatively common congenital malformation of the urinary tract. This condition appears to have a multifactorial cause, with a combination of genetic and environmental factors. Further research is needed to show a better understanding of these risk factors. PMID- 11920334 TI - Expression of glomerular plasminogen activator inhibitor type 1 in glomerulonephritis. AB - Plasminogen activator inhibitor type 1 (PAI-1) and tissue plasminogen activator (tPA) are the major regulators of plasmin generation. Glomerular PAI-1/tPA balance is involved in extracellular matrix turnover, as well as fibrin deposition in glomeruli. Renal biopsy specimens were obtained from 80 patients with either primary or secondary glomerulonephritis (10 patients, minimal change nephrotic syndrome; 6 patients, focal segmental glomerulosclerosis [FSGS]; 10 patients, membranous nephropathy [MN]; 24 patients, mesangial proliferative glomerulonephritis; 15 patients, lupus nephritis; 14 patients, diabetic nephropathy; and 1 patient, membranoproliferative glomerulonephritis). We quantified glomerular PAI-1 and tPA messenger RNA (mRNA) by competitive polymerase chain reaction. We also determined PAI-1 mRNA localization by in situ hybridization. Glomerular PAI-1 mRNA levels in patients with FSGS and MN were significantly greater than those of controls. There was a sixfold increase in PAI 1-tPA mRNA ratio in patients with MN compared with the control group. In addition, glomerular PAI-1 mRNA level correlated with level of proteinuria. Conversely, there was no difference in tPA mRNA levels among types of glomerulonephritis. These results suggest that suppressed glomerular fibrinolytic and proteolytic activity may be associated with the pathogenesis of glomerulonephritis, especially in FSGS and MN. PMID- 11920335 TI - Bucillamine induces membranous glomerulonephritis. AB - A variety of renal histopathologic lesions, such as amyloidosis, mesangial proliferative glomerulonephritis, and membranous glomerulonephritis (MGN), are associated with rheumatoid arthritis (RA). Bucillamine (BCL), a disease-modifying antirheumatic drug, has a chemical structure and side-effect profile similar to that of d-penicillamine, which can induce MGN in RA. There are a few reports of MGN occurring in association with BCL treatment. However, lacking detailed analyses of immunoglobulin deposition in glomerular lesions, these studies did not elucidate the pathogenesis of BCL-induced MGN. We evaluated seven biopsy specimens from six patients with RA who had undergone BCL treatment with a mean BCL dose of 72.5 g before the appearance of proteinuria. Light microscopic evaluation showed mild to moderate mesangial proliferation. Two biopsy specimens showed spikes along glomerular capillary walls. Granular deposition of immunoglobulin G (IgG) along glomerular capillary walls was seen in all cases, and five specimens showed deposition of IgG2 and/or IgG3 components, in addition to IgG4. Furthermore, subepithelial dense deposits were distributed segmentally in four biopsy specimens on electron microscopy. IgG4, reported to be the predominant IgG subclass deposited, is distributed diffusely in idiopathic MGN. Thus, there were obvious differences between BCL-induced and idiopathic MGN in regard to both IgG subclasses deposited and deposition pattern within the glomerulus. Because IgG3 has the strongest affinity for C1q, these findings suggest that BCL-induced MGN activates the classical pathway more efficiently than idiopathic MGN and that the pathogenesis is different between these two diseases. PMID- 11920336 TI - Different patterns of renal damage in type 2 diabetes mellitus: a multicentric study on 393 biopsies. AB - The frequency of various types of renal changes in patients with type 2 diabetes is not clearly defined in the literature. Reported discrepancies likely are caused by ethnic and geographic factors. However, policies used in nephrological centers for the selection of patients to undergo renal biopsy also may have an influence. The present study reports 393 renal biopsies in patients with type 2 diabetes performed in a group of centers in northwestern Italy using different (restricted [CRPs] or unrestricted [CUP]) biopsy policies. On the basis of light microscopic, immunofluorescence, and ultrastructural findings, cases were subdivided into three classes characterized by the presence of diabetic glomerulosclerosis (class 1), prevailing vascular (arterioarteriolosclerotic) and ischemic glomerular changes (class 2), other glomerulonephritides superimposed on diabetic glomerulosclerosis (class 3a), or glomerulonephritides without the presence of diabetic glomerulosclerosis (class 3b). Although no significant differences were found for class 2 (detected in 15% and 16% of patients from CRPs and the CUP, respectively), the frequency of the other two classes was strongly biased by the biopsy policy. Class 1 was found in 29% and 51% of cases, and class 3 in 57% and 33% of cases from CRPs and the CUP, respectively. Moreover, class 3a was more common (67%) in the CUP, and class 3b (78%) in CRPs. Our findings may explain conflicting data from the literature and the influence that type of adopted biopsy policy may have on an epidemiological evaluation. This study helps clarify the frequency of renal changes in patients with type 2 diabetes and suggests more extensive use of renal biopsy to obtain reliable prognostic indications and plan a rational therapeutic approach. PMID- 11920337 TI - Slowing the progression of chronic renal failure: economic benefits and patients' perspectives. AB - Because of the predicted increase in end-stage renal disease (ESRD) incidence (projected increase from 1998 to 2010; 86,825 to 172,667), prevalence (projected increase from 1998 to 2010; 326,217 to 661,330), and cost (total cost based on 1998 ratio of Medicare versus non-Medicare cost; $16.74 billion in 1998 to $39.35 billion in 2010), a cohesive national effort is needed to develop strategies to slow the progression of chronic renal failure (CRF). The question arises to how much reduction in the progression of CRF would lead to a meaningful decrease in the prevalence and cost of ESRD. There are no objective data that show the economic impact of slowing the progression of CRF. We developed a mathematical model to assess the economic impact of decreasing the progression of CRF by 10%, 20%, and 30%. US Renal Data System (USRDS) projections were used to model the rate of increase in ESRD incidence and prevalence. Glomerular filtration rate (GFR) at the initiation of ESRD therapy and cost per patient-year were based on USRDS data. The average decline in GFR in subjects with CRF was estimated to be 7.56 mL/min/y. All dollar savings reflect 1998 costs, discounted for the future at 3% per annum. We also determined how much slowing of the progression of CRF is important from patients' perspectives by means of a written questionnaire (which inquired about willingness to go on a restricted diet, take six extra medications per day, and make six extra office visits per year) and calculation of the pre ESRD time gained for different degrees of reduction in the progression of CRF. If the rate of decline in GFR decreased by 10%, 20%, and 30% after December 31, 1999, in all patients with GFRs of 60 mL/min or less, cumulative direct healthcare savings through 2010 would equal approximately $18.56, $39.02, and $60.61 billion, respectively. For a 10%, 20%, and 30% decrease in the rate of decline in GFR in all patients with a GFR of 30 mL/min or less, estimated cumulative savings through 2010 equal $9.06, $19.98, and $33.37 billion, respectively. Responses to the questionnaire showed that approximately 79% of subjects with CRF (n = 113) perceived a few weeks' dialysis-free period significant (P < or = 0.0001), a period corresponding to a 10% reduction in the rate of decline in GFR. Our data suggest that the cumulative economic impact of slowing the progression of CRF, even by as little as 10%, would be staggering. They provide strong support for the development and implementation of intensive reno-protective efforts beginning at the early stages of chronic renal disease and continued throughout its course. PMID- 11920338 TI - Renal function after aortic stent-grafting including coverage of renal arterial ostia. AB - Considerable experience has been gained with endoluminal stent-graft techniques for the treatment of infrarenal aortic aneurysms. The inclusion of uncovered suprarenal stents eases the treatment of cases with a short, angulated, or bottlenecked proximal neck. However, few data have been published on the effect of a stent-graft crossing renal artery ostia on long-term renal function. We therefore retrospectively analyzed the influence of intra-aortic stent-grafts, including suprarenal fixation on long-term renal function. Thirty-nine patients who received a Gianturco Z stent across renal artery ostia as part of a made-to measure stent-graft treating an infrarenal aortic aneurysm were evaluated at 6 (all patients) and 30 months (15 patients) after surgery. Renal function evaluation included serum creatinine level, creatinine clearance using the Cockcroft-Gault formula, and renal tomography. Initial technical success was achieved in all patients. There were no minor or major complications. Mean serum creatinine levels were 1.16 +/- 0.25, 1.12 +/- 0.28, and 1.24 +/- 0.25 mg/dL before and 6 and 30 months after endoluminal stent-graft performance, respectively (P = not significant [NS]). Mean creatinine clearances were 60.3 +/- 19.7, 63 +/- 23, and 60.6 +/- 17.5 mL/min before and 6 and 30 months after endoluminal stent-graft performance, respectively (P = NS). Only two patients (5%) had a decrease in glomerular filtration rate greater than 20% at 6 months. Both patients had renal insufficiency before endovascular grafting. Renal function was stable in all patients at 30 months' follow-up. We suggest that renal function is unaffected by coverage of renal arterial ostia with Gianturco Z stents. PMID- 11920339 TI - Intrafamilial phenotype variability in nephrogenic diabetes insipidus. AB - X-Linked nephrogenic diabetes insipidus (NDI), which accounts for 90% of inherited cases of NDI, is caused by mutations in the AVPR2 gene that encodes the arginine vasopressin (AVP) receptor type 2 (V2R). The V2R mediates the antidiuretic action of AVP in principal cells of the collecting duct. To date, only three AVPR2 mutations (P322S, D85N, and G201D) have been associated with a mild NDI phenotype, and intrafamilial phenotype variability has not been reported in affected males. We describe a novel Belgian family with X-linked NDI caused by substitution of a histidine for an arginine at position 137 (R137H) of AVPR2. This mutation has been identified in two brothers and their mother. The R137H mutation results in a failure of V2R to stimulate adenylate cyclase and has been associated consistently with severe NDI and the inability to increase urinary osmolality to greater than plasma osmolality during water deprivation and/or infusion of 1-desamino-8-d-arginine vasopressin. Detailed examination of the two affected brothers showed the typical NDI phenotype in the 45-year-old proband, whereas a milder clinical phenotype associated with significant urinary concentrating ability during water deprivation was documented in the 33-year-old brother. Thus, in this family, the R137H mutation is associated with either a mild or severe NDI phenotype. Mechanisms that might account for these findings include genetic and/or environmental modifiers. PMID- 11920340 TI - Roles of host and bacterial virulence factors in the development of upper urinary tract infection caused by Escherichia coli. AB - Aims of this study are to identify host and Escherichia coli virulence factors associated with upper urinary tract infection (UTI) by comparing them with those for lower UTI and determining the association between major predisposing host factors for upper UTI and urovirulence genes for E coli. Host factors and urovirulence genes of E coli associated with bacteremia in patients with upper UTI and their interactions also were studied. One hundred thirty-nine adult patients who fulfilled clinical diagnostic criteria for upper (n = 81) or lower UTI (n = 58) caused by E coli between January 1997 and December 1999 were retrospectively enrolled into this study. Old age (> or =60 years), male sex, diabetes with poor blood glucose control (ie, glycosylated hemoglobin A1C > or = 8.1%), immunosuppression, and urinary tract obstruction were more frequently associated host factors for patients with upper UTI than for those with lower UTI. Using polymerase chain reaction, the papG class II allele was detected more frequently for E coli strains isolated from patients with upper UTI than for those from patients with lower UTI (85% versus 52%; P < 0.0001). Multivariate analysis showed that diabetes with poor blood glucose control, immunosuppression, urinary tract obstruction, and papG class II allele were independently associated with upper UTI. For patients without these three predisposing host factors, the prevalence of papG class II allele was significantly greater in those with upper UTI than those with lower UTI. However, the papG class II allele was less prevalent in strains isolated from patients with upper UTI with urinary tract obstruction or with two of the three predisposing host factors. In addition, both univariate and multivariate analyses showed that old age and papG class II allele were risk factors for the development of E coli bacteremia in patients with upper UTI. In conclusion, both host and E coli virulence factors contribute to the development of upper UTI, and less virulent strains can cause upper UTI in hosts with predisposing factors. PMID- 11920341 TI - Optimal strategies for modeling the reciprocal of creatinine versus time in renal transplant recipients and patients with native chronic renal disease. AB - Few studies have evaluated the least squares (LS) or alternative regression methods to estimate loss of renal function using the reciprocal of creatinine over time in renal transplant recipients or have compared their performances in patients with chronic renal insufficiency (CRI). We evaluated the LS and alternative methods using the proportion of explained variance, measured by R2, and prediction of the final creatinine level. The study included two groups of transplant recipients: (1) patients who developed graft failure (FAIL; n = 31) and (2) patients with an episode of biopsy-proven acute rejection with or without subsequent renal function loss (BXAR; n = 98) and a third group of individuals with CRI (n = 28). The LS method performed poorly in both transplant groups (mean R2 range, 0.35 to 0.44; 32% to 45% with final creatinine(actual) - creatinine(predicted) +/- 0.05 mg/dL), but better in the CRI group (mean R2, 0.57; 75% with final creatinine(actual) - creatinine(predicted) +/- 0.05). The best alternative was the two-phase regression line after exclusion of outliers, which provided similar results across the FAIL, BXAR, and CRI groups (mean R2, 0.71, 0.64, and 0.73; 81%, 84%, and 96% with final creatinine(actual) - creatinine(predicted) +/- 0.05, respectively; P = not significant) and had the best performance in patients with greater nadir creatinine values. The LS method is not suited to measure kidney function loss in renal transplant recipients. Use of the two-phase regression line after exclusion of outliers is a more reliable method in renal transplant recipients, especially patients with impaired baseline function, and has results similar to those of patients with CRI. PMID- 11920342 TI - A thirty percent chronic decline in inverse serum creatinine is an excellent predictor of late renal allograft failure. AB - We previously reported that the percentage of change in inverse serum creatinine (Delta1/Cr) was the best of several time-dependent serum creatinine-derived predictors of renal allograft failure in patients not administered cyclosporine (CsA). To further validate the utility of Delta1/Cr, we collected creatinine levels (mean, 90.7 +/- 45.2 creatinine measurements) in 100 patients treated for 6.7 +/- 5.9 years with CsA. We also validated Delta1/Cr using a limited creatinine-sampling strategy, then performed multivariate Cox proportional hazards analysis of 1,663 transplantations. A time-dependent covariate determined by the date of first chronic decline (excluding creatinine levels from periods of acute rejection) in Delta1/Cr to less than -30% of baseline similarly was predictive of graft failure in 101 patients treated without CsA (relative risk, 5.04; 95% confidence interval, 2.18 to 11.6; P = 0.0002) and 100 patients treated with CsA (relative risk, 5.02; 95% confidence interval, 2.50 to 10.1; P < 0.0001). A limited creatinine-sampling strategy (measured at 1 week, 1, 3, 6, 12, 18, 24, and 36 months, and each year thereafter) reduced the ability of Delta1/Cr less than -30% to predict graft failure. In 1,663 patients, Delta1/Cr less than 30% first occurred a median of 1.0 years posttransplantation (n = 792 of 1,663 patients) and 3.0 years before graft failure (n = 478 of 897 patients with graft failure). In a multivariate model (n = 1,663) that included baseline function, acute rejection, and other covariates, Delta1/Cr less than -30% was a strong independent predictor of graft failure (relative risk, 2.56; 95% confidence interval, 2.12 to 3.09; P < 0.0001). Thus, Delta1/Cr less than -30% is an excellent predictor of graft failure that is similarly predictive in patients treated with and without CsA. A limited sampling strategy for creatinine diminishes, but does not negate, the usefulness of Delta1/Cr less than -30%. PMID- 11920343 TI - Renal transplant in patients with Alport's syndrome. AB - We evaluated 52 renal grafts transplanted into 41 patients with a pretransplantation diagnosis of Alport's syndrome. Overall 1-, 5-, and 10-year patient and graft survival rates were 95.1%, 90.2%, and 80.5% and 86.8%, 66%, and 45.3%, respectively. Although 14% of renal graft biopsy specimens examined with immunofluorescent microscopy showed linear glomerular basement membrane (GBM) immunoglobulin G deposits, only 1 of 41 patients (2.4%) or 52 grafts (1.9%) developed posttransplantation anti-GBM disease. The incidence of anti-GBM disease was 3.1% (1 of 32 patients) in a subgroup of male transplant recipients. Our analysis suggests that the incidence of anti-GBM disease in transplant recipients with Alport's syndrome is less than previously reported. In addition, it does not appear that HLA-DR alleles, which predispose to the development of anti-GBM disease in native kidneys, have a role in transplant recipients with Alport's syndrome posttransplantation. However, immunosuppression level may have a pathophysiological role in the development of anti-GBM disease. The majority of grafts in transplant recipients with Alport's syndrome failed because of chronic allograft nephropathy (69% of grafts) and acute rejection (22% of grafts). A history of previous acute rejection was the only factor that significantly affected graft outcome. PMID- 11920344 TI - Placental imbalance of vasoactive factors does not affect pregnancy outcome in patients treated with Cyclosporine A after transplantation. AB - Endothelin-1 (ET-1) and nitric oxide (NO) have been suggested to have a focal role in the regulation of placental and fetal growth. Cyclosporine A (CsA) has been shown to strongly modulate ET-1 and NO synthesis and thus has the potential to affect fetal growth and maternal state. Eleven CsA-treated female kidney transplant recipients were recruited. Fourteen healthy pregnant women served as controls. Placental expression of ET-1 and tissue factor (TF) was evaluated by in situ hybridization, and NO synthase (NOS) was evaluated by staining with the reduced form of nicotinamide-adenine dinucleotide phosphate (NADPH)-diaphorase and in situ hybridization. Kidney transplant recipients showed a marked reduction in NADPH-diaphorase staining, as well as endothelial constitutive NOS (ecNOS) messenger RNA, whereas inducible NOS expression was unchanged. Normal placenta showed a strong positive ET-1 signal along the endothelium of uteroplacental arteries within the basal plate, which increased markedly in decidua of transplant recipients. Thus, transplant recipients showed a remarkable alteration in ET-1/ecNOS balance without alteration in fetal growth or maternal renal function. Next, we explored the state of placental endothelial cell activation downstream from vasoactive factors by evaluating TF gene expression. Transplant recipients did not show modification of TF transcript compared with healthy pregnant women. CsA potently affected the placental ET-1/ecNOS vasoactive balance. Nevertheless, newborns from transplant recipient mothers were appropriate for gestational age, and transplant recipients did not show systemic hypertension or impending renal damage. It is suggested that CsA may blunt the activation of endothelial cells and priming of endothelial-derived substances, which possibly lie downstream from the cited vasoactive agents. PMID- 11920346 TI - Comparison of mortality and intermediate outcomes between medicare dialysis patients in HMO and fee for service. AB - End-stage renal disease (ESRD) is the only disease entitlement for Medicare; therefore, most patients with ESRD have Medicare coverage. Patients with ESRD are prohibited by law from enrolling in health maintenance organizations (HMOs), the only group prohibited within Medicare. However, they may remain in an HMO if they enrolled in such a plan before their kidneys failed. Thus, it is possible to compare patients with ESRD in HMOs with those in fee-for-service (FFS) plans. To determine whether mortality, transplantation rates, and intermediate outcomes differed between Medicare ESRD beneficiaries enrolled in HMO versus FFS providers, a retrospective cohort analysis was performed of patients with ESRD from three Health Care Financing Administration data sets containing administrative and outcome information for Medicare ESRD beneficiaries from 1990 to 1998. On December 31, 1998, a total of 278,510 prevalent patients with ESRD were in FFS plans, and 18,332 patients were in HMOs. HMO patients were older and more likely to be white and male and have diabetes mellitus and comorbid cardiovascular conditions than FFS patients. Unadjusted 2-year survival rates were 48.4% and 49.3% for FFS and HMO patients, respectively. In a multivariate model, HMO status had no significant effect on mortality, which was greater with older age, male sex, and white race. In 1998, unadjusted renal transplantation rates were 23.5% and 15.5% for FFS and HMO patients, respectively; age adjustment abrogated the apparent difference. For FFS and HMO patients, adequate hemodialysis was delivered to 72% and 82%, and 56% and 62% had hematocrits greater than the benchmark, respectively. There was no statistical difference in these outcomes in multivariate comparison. In conclusion, care by HMO for patients with an expensive chronic illness can achieve outcomes similar to those for FFS patients. Claims of poorer care and worse outcomes for patients with ESRD enrolled onto an HMO, an argument used to justify continued prohibition against widespread participation by patients with ESRD, are not supported. PMID- 11920345 TI - United States Renal Data System assessment of the impact of the National Kidney Foundation-Dialysis Outcomes Quality Initiative guidelines. AB - Since 1989, significant efforts have focused on improving the care of dialysis patients in the United States. Numerous organizations have developed clinical practice guidelines; however, few guidelines have received the broad support given to the National Kidney Foundation-Dialysis Outcomes Quality Initiative (DOQI). These guidelines, independently developed from an extensive review of the literature, include sections on dialysis adequacy, anemia treatment, and vascular access. To assess the impact of these guidelines on clinical practice, we evaluated data on hematocrits, recombinant human erythropoietin dosing, hemodialysis adequacy, and simple fistula and dialysis catheter utilization using Medicare dialysis provider claims and Medicare Part B physician services. Hematocrits have increased steadily, with the exception of the period when the Hematocrit Measurement Audit was in effect. After cancellation of the policy, hematocrits increased to the midpoint of the DOQI target range (34.4%). Although the level of dialysis therapy has stabilized, with the average urea reduction rate of 68% to 69.9% in 1997 to 1999 being slightly greater than the DOQI target of 65% or greater, geographic variability is apparent. Simple fistula placement rates increased by 45% during the pre-DOQI and post-DOQI period from 1994 to 1999. The use of temporary catheters decreased, whereas placement of permanent catheters has increased, which may reflect recommended practice guidelines. Although it appears that clinical practice guidelines have improved the clinical care of dialysis patients, considerable regional variations in care across the country should be given significant attention. PMID- 11920347 TI - Burden and quality of life of caregivers for hemodialysis patients. AB - The aim of this study is to describe the characteristics of caregivers of chronic hemodialysis patients, assess their perceived burden and health-related quality of life, and investigate factors influencing this burden. We studied 100 hemodialysis patients and their respective primary caregivers for more than 4 months, measuring quality of life by the Medical Outcomes Survey 36-Item Short Form Health Survey (SF-36). Subjective burden on caregivers was assessed by the Caregiver Burden scale (score range, 1 to 4; higher values indicate a greater effect). The majority of caregivers were women (84%), married (66%), with a mean age of 46 +/- 2 (SE) years, and of low socioeconomic level. Their main types of relationship with patients were wives (38%) and sons or daughters (27%). Caregiver Mental Health and Vitality were the most affected emotional dimensions on the SF-36 (mean scores, 64.4 +/- 1.8 and 66.6 +/- 1.7, respectively). Mean score of total burden experienced was 2.07 +/- 0.05. Multiple regression analysis showed that independent and significant predictors of burden were Mental Health of the caregiver (R2 = 24%), Vitality of the patient (R2 = 10%), type of relationship of the caregiver (female spouse) (R2 = 5%), and Pain of the caregiver (R2 = 3%). Caregivers of hemodialysis patients may experience a significant burden and an adverse effect on their quality of life. Emotional aspects of caregivers (particularly female spouses) and patients are important predictors of burden. Social support and psychological interventions should be considered to improve caregiver life and patient outcomes. PMID- 11920348 TI - Hemodialysis access assessment with intravascular ultrasound. AB - Reliable identification and treatment of specific hemodialysis access complications may improve access patency and result in significant cost reduction. Angiography is the gold standard for the evaluation of vascular access; however, it has significant limitations. Intravascular ultrasound (IVUS) is a relatively new technique capable of detecting subtle vascular abnormalities. To investigate the safety, feasibility, and accuracy of IVUS imaging to detect hemodialysis access complications, including stenoses, graft deterioration, and thrombus, we performed 31 IVUS imaging studies in 22 hemodialysis patients. Nineteen studies were performed in the dialysis unit, and 12 studies in the angiography suite. The IVUS catheter was inserted into the graft through the access used for hemodialysis. Findings of 21 studies (17 patients) imaged on the same day by both angiography and IVUS were compared. Grafts and vessels were successfully imaged using IVUS in 29 of 31 studies. There were no adverse effects caused by IVUS. Angiography assessed 17 of 54 vessel segments as normal versus 9 of 54 segments by IVUS (P < 0.001). Angiography detected lesions in 25 segments as opposed to 33 segments by IVUS (P < 0.001). A thrombus was detected in 32 of 54 vessel segments by IVUS, but in only 1 of 54 segments by angiography (P < 0.001). In conclusion, IVUS imaging is feasible and safe to assess hemodialysis access in the angiographic suite and dialysis unit. IVUS detected more vascular abnormalities than angiography. IVUS may be a useful independent imaging and screening modality in the assessment of dialysis access complications, which may help increase graft patency and reduce cost. PMID- 11920349 TI - Salvage of poorly developed arteriovenous fistulae with percutaneous ligation of accessory veins. AB - Many arteriovenous (AV) fistulae fail to achieve an adequate blood flow or size for successful cannulation because of accessory veins. We describe a simple technique to ligate accessory veins that does not require a surgical incision. In this retrospective study, 17 end-stage renal disease patients underwent ligation of accessory veins of poorly developed AV fistulae. There were 14 men and 3 women, and their average age was 50 +/- 13 years. There were 14 radiocephalic and 3 brachiocephalic fistulae. After identifying accessory veins with a fistulogram, two nonabsorbable 2-0 polypropylene (Prolene) sutures were placed percutaneously around each accessory vein in proximity to the AV fistula. Successful ligation was confirmed with a repeat fistulogram. This procedure was undertaken after 4 +/ 3 months following surgical placement. Successful maturation was defined as adequate blood flow to support effective hemodialysis and adequate caliber to allow for repeated cannulation with a 15G or 16G needle. Of 17 AV fistulae, 15 (88%) successfully matured 1.7 +/- 1 month (range, 0.3 to 6 months) after the procedure. The average number of accessory veins ligated was 1.7 +/- 0.8 (range, 1 to 3). All AV fistulae that matured after ligation of accessory veins were functioning at 44.5 +/- 12 weeks after first use. A technique for salvaging nonmaturing AV fistulae not requiring surgical cutdown for ligation of accessory veins is described. AV fistulae mature quickly after ligation of accessory veins. This is a rapid and safe procedure that can increase the prevalence of AV fistulae. PMID- 11920350 TI - Exercise during hemodialysis decreases the use of antihypertensive medications. AB - Most hemodialysis patients require antihypertensive therapy. Aerobic exercise has been suggested as a nonpharmacologic treatment for hypertension in many patient populations, including those with chronic renal failure. To test the effectiveness of this therapy in an outpatient long-term hemodialysis clinic, the hemodialysis staff instituted a stationary cycling program during dialysis and offered the program to all patients (n = 107). Forty patients agreed to participate, and 35 nonexercising patients served as controls. Patients performed stationary cycling during each hemodialysis treatment. Predialysis blood pressures, postdialysis blood pressures, and antihypertensive medication use were recorded during a 6-month period. Costs of the medication were analyzed at the end of the study. Of participants, 24 (60%) completed 6 months of exercise with a mean increase in total cycling time from 16.9 min/session to 45.5 min/session. No serious adverse events were reported. Predialysis and postdialysis blood pressures were not statistically different between the two groups at month 0 or month 6, but 13 (54%) in the exercise group had a reduction in antihypertensive medication versus 4 (12.5%) in the control group (P = 0.008). The average relative benefit of exercise was a 36% reduction in antihypertensive medications (P = 0.018) with an average annual cost savings of $885/patient-year (P = 0.005) in the exercise group. Stationary cycling is safe during hemodialysis and can lead to significant reductions in blood pressure medication use and cost savings, justifying the initial capital cost of equipment and small incremental increase in staff time. PMID- 11920351 TI - Value of corrected QT interval dispersion in identifying patients initiating dialysis at increased risk of total and cardiovascular mortality. AB - Cardiovascular disease remains the most common cause of premature death in end stage renal disease (ESRD). Although several predictors of cardiac death have been reported, identifying individuals most at risk remains difficult. Previous studies in nonuremic populations have associated cardiac mortality, in particular sudden death, with increased QT dispersion (QTd); defined as the difference between the maximal and minimal QT interval on a standard electrocardiogram. The present study aimed to determine the prognostic value of QTd and corrected QTd (QTdc) in predicting total, cardiovascular, and arrhythmia-related mortality in ESRD patients initiating dialysis. The study was a retrospective cohort of adult ESRD patients starting peritoneal dialysis or hemodialysis between 1990 and 1994. Statistical analysis was by Cox proportional hazard modeling and Kaplan-Meier analysis. Primary study endpoints were total, cardiovascular, and arrhythmia related mortality. Nonfatal cardiovascular events were a secondary endpoint. A total of 147 patients were studied for a period of 5 to 9 years. In Cox modeling, QTdc was an independent predictor of total (relative risk [RR] = 1.53; difference for RR = 50 msec; P = 0.0001) and cardiovascular mortality (RR = 1.57; difference for RR = 50 msec; P = 0.028) and trended toward arrhythmia-related mortality (P = 0.061). Total mortality also was predicted independently by lack of renal transplantation, radiographic cardiomegaly, and predialysis serum albumin. In multivariate analysis, QTdc was associated weakly with serum calcium, mean QT interval, and presence of diabetes mellitus. QTdc may be a useful marker for identifying dialysis patients at an increased risk for overall and cardiovascular mortality. PMID- 11920352 TI - Bone density and heel ultrasound testing do not identify patients with dialysis dependent renal failure who have had fractures. AB - Patients with dialysis-dependent renal failure are at increased risk for low trauma fractures. However, the optimal means of identifying patients at high risk for fracture is not known. We assessed the association between fracture history and two tests of bone mineral density (dual x-ray absorptiometry [DEXA] and calcaneal ultrasound) among patients with hemodialysis-dependent renal failure. We evaluated 71 men and 33 women aged 55 years or older who had been receiving hemodialysis for at least 1 year. All patients underwent spinal radiography, DEXA of the hip and lumbar spine, and calcaneal ultrasonography. We assessed risk factors for low-trauma fractures by questionnaire and medical chart review. Of patients, 52% had a fracture on spinal radiographs or a history of a low-trauma fracture, 69% had osteopenia by DEXA, and 26% had a low heel ultrasound measurement. Neither DEXA nor calcaneal ultrasound was associated with fracture history, however. Our findings indicate that fractures among patients with dialysis-dependent renal failure are common. Tests of bone strength do not adequately identify patients with a history of fractures. Prospective studies to determine the optimal method of identifying patients with dialysis-dependent renal failure at high risk for fracture are needed. PMID- 11920353 TI - Predialysis versus postdialysis hematocrit evaluation during erythropoietin therapy. AB - American guidelines for the management of renal anemia by recombinant human erythropoietin (rHuEPO) recommend collecting a predialysis blood sample to evaluate hemoglobin (Hb) and hematocrit (Hct) levels in hemodialysis patients. Although a predialysis blood sample is appropriate for evaluating when to start rHuEPO treatment, the same sample would not be appropriate for evaluating the target Hb/Hct to be maintained, particularly when normal or near-normal values are pursued. We measured the degree of intradialytic and extradialytic variation of Hb, Hct, and body weight in 68 stable hemodialysis patients on maintenance subcutaneous rHuEPO treatment. Hb and Hct concentrations were determined before and after dialysis. In 16 patients, Hb and Hct concentrations also were assessed 24 hours after the end of dialysis. Predialysis versus postdialysis Hb and Hct concentrations for all patients were 10.5 +/- 1.3 g/dL versus 11.5 +/- 1.3 g/dL (P < 0.0001) and 32 +/- 4% versus 35 +/- 4% (P < 0.0001). The intradialytic percent variation (%Delta) of Hct and body weight were 10 +/- 6% and -6.3 +/- 3.5%. There was a close inverse correlation between %Delta of Hct and Hb and %Delta of body weight (P < 0.0001). In patients with body weight losses 2.5 kg or more per session, the mean %Delta of Hct was 12 +/- 7%. In the 16 patients studied 24 hours after the end of the dialysis session, Hct and Hb values remained significantly higher compared with the predialysis levels (P < 0.001), suggesting a slow reequilibration of the intravascular volume in the first 24 hours after hemodialysis. For these reasons, predialysis samples for monitoring the target Hb and Hct levels in patients treated by rHuEPO should be considered with caution. PMID- 11920354 TI - Hemodialysis-activated granulocytes at the site of interstitial inflammation. AB - It is not known to what extent intravascular phenotypic alterations in adhesion molecule expression induced by hemodialysis influence the recruitment of granulocytes and their ability to up-regulate CD11b at the local site of inflammation in the interstitium. We used a skin suction chamber technique to address this issue. Two skin blisters were raised on the forearm of eight hemodialysis patients and eight healthy subjects, and blister exudate was collected (time 0). The two blisters were stimulated with buffer (intermediate inflammation) or autologous serum (intense inflammation). Then the patients underwent cuprophane hemodialysis for 4 hours. Ten hours after start of dialysis, the exudate was aspirated from each chamber. Granulocyte count and surface expression of CD11b and CD62L were analyzed in samples from peripheral blood and blister exudate by flow cytometry. Granulocytes from healthy blood donors were incubated in blister fluid from patients and healthy subjects to determine the local chemotactic activity in terms of CD11b up-regulation. The expression of CD11b increased fourfold and CD62L decreased simultaneously in patients and healthy subjects when cells transmigrated to the unstimulated blister at time 0. At the site of intermediate inflammation, granulocytes from patients had a significantly lower capacity to mobilize CD11b compared with cells from healthy subjects (P < 0.001). At the site of intense interstitial inflammation, granulocytes from patients had the capacity to mobilize the receptor and reached values close to those obtained in healthy subjects (P = 0.079). The blister exudate from patients had a similar (at time 0 and intermediate inflammation) or higher (intense inflammation; P < 0.05) capacity to up-regulate CD11b on granulocytes in vitro compared with blister exudate from healthy subjects. Granulocytes from hemodialysis patients seem to require a more intense chemotactic stimulus to up-regulate CD11b at the local site of inflammation in the interstitium compared with corresponding cells from healthy subjects despite the fact that cells transmigrate in a milieu that contains chemotactic factors with an equal or higher capacity to up-regulate CD11b. Granulocytes in hemodialysis patients seem to be more refractory to inflammatory stimuli in the interstitium. PMID- 11920355 TI - Efficacy and safety of a 7.5% icodextrin peritoneal dialysis solution in patients treated with automated peritoneal dialysis. AB - In a randomized, prospective, multicenter study, we compared the safety, efficacy, and metabolic effects of a 7.5% icodextrin solution (Extraneal) with a 2.27% glucose solution for long dwell exchanges in patients undergoing automated peritoneal dialysis. Thirty-nine stable patients on automated peritoneal dialysis were randomized to receive either icodextrin (n = 20) or glucose 2.27% solution (n = 19). The study included a 2-week baseline period followed by a 12-week icodextrin treatment phase and a 2-week follow-up period when switching back to glucose. The average net ultrafiltration during the long dwell period was 278 +/- 43 mL/d for the icodextrin group and -138 +/- 81 mL/d for the control group (P < 0.001). The higher ultrafiltration volume with icodextrin was associated with higher creatinine (2.59 +/- 0.09 mL/min versus 2.16 +/- 0.11 mL/min) and urea (2.67 +/- 0.09 mL/min versus 2.28 +/- 0.12 mL/min) peritoneal clearances for the long dwell (both P < 0.001). Ultrafiltration rate per mass of carbohydrate absorbed was +5.2 +/- 1.2 microL/min/g in the icodextrin group and -5.5 +/- 2.8 microL/min/g in the glucose group (P < 0.001). In the icodextrin group, there was a decrease in serum sodium and chloride compared with baseline (P < 0.01). Total dialysate sodium removal increased in the icodextrin group from 226.7 mEq to 269.6 mEq (week 12, P < 0.001). Serum alpha-amylase activity decreased from 103 U/L to 16 U/L (P < 0.001). The total icodextrin plasma levels reached a steady state concentration of 6,187 +/- 399 mg/L after 1 week of treatment. Urine volume and residual renal function were not specifically affected by icodextrin compared with glucose. None of the laboratory changes resulted in any reported clinically meaningful side effect. Icodextrin produced increased, sustained ultrafiltration during the long dwell period, increasing (convective) peritoneal clearance and sodium removal in automated peritoneal dialysis patients. PMID- 11920356 TI - Can a reduction in extracellular fluid volume result in increased serum albumin in peritoneal dialysis patients? AB - Serum albumin predicts survival in end-stage renal failure. The literature has emphasized the dependence of albumin on inflammation. We previously proposed an independent relationship with overhydration. To date, there is no proven therapy that increases serum albumin in dialysis patients. We investigated whether decreasing dry weight increases serum albumin in peritoneal dialysis patients. Twenty-one subjects (15 continuous ambulatory peritoneal dialysis patients and 6 continuous cycled peritoneal dialysis patients) were assessed at 0 and 4 weeks. Body weight, blood pressure, 4-site skin-fold thickness, subjective global assessment score, and use of antihypertensive medication were recorded. Fluid samples were collected for estimation of serum albumin, C-reactive protein, and 24-hour dialysate and urine volume. The extracellular fluid volume was estimated by multiple-frequency bioelectric impedance. Between the 0- and 4-week assessments, dialysis prescription was altered to increase the daily ultrafiltrate volume (1.00 +/- 0.71 L to 1.29 +/- 0.75 L; P = 0.04). Significant decreases were seen in body weight (70.7 +/- 12.1 kg to 69.9 +/- 12.2 kg; P = 0.0002), extracellular fluid volume (16.2 +/- 3.3 L to 15.5 +/- 3.2 L; P = 0.0006), systolic blood pressure (147.7 +/- 26.3 mm Hg to 124.2 +/- 20.9 mm Hg; P < 0.0001), diastolic blood pressure (84.2 +/- 14.5 mm Hg to 76.6 +/- 11.7 mm Hg; P = 0.01), and number of antihypertensive drugs (1.9 +/- 1.0 to 1.6 +/- 1.1; P = 0.02). Serum albumin increased (34.6 +/- 4.1 g/L to 35.9 +/- 3.6 g/L; P = 0.01). C-reactive protein did not change. Decreasing dry weight in peritoneal dialysis by an increase in ultrafiltration volume results in a decreased extracellular fluid volume, which is reflected by an improvement in blood pressure control. This improvement in blood pressure was associated with an increase in serum albumin. This finding strengthens the argument that overhydration is a cause of hypoalbuminemia in peritoneal dialysis and offers a therapeutic option in management of this patient group. PMID- 11920357 TI - Oral interferon-alpha treatment of mice with cryoglobulinemic glomerulonephritis. AB - Cryoglobulins are associated with the development of a membranoproliferative glomerulonephritis, often referred to as cryoglobulinemic glomerulonephritis, particularly in the setting of hepatitis C virus infection. Parenteral interferon alpha (IFN-alpha) commonly is used therapeutically in humans with cryoglobulinemic glomerulonephritis. We tested the therapeutic impact of oral IFN alpha treatment in thymic stromal lymphopoietin (TSLP) transgenic mice, a strain that develops mixed cryoglobulinemia with glomerulonephritis closely resembling the disease that occurs in humans. A total of 41 female mice were treated for 21 days with daily ingestion of either 500 IU of Universal type I IFN or placebo. The studied groups included TSLP transgenic mice treated with IFN-alpha (n = 13), TSLP transgenic mice treated with placebo (n = 13), wild-type mice treated with IFN-alpha (n = 5), and wild-type mice treated with placebo (n = 10). A total of 39 mice completed the study; two TSLP transgenic mice treated with IFN-alpha died during the study period. Placebo-treated TSLP transgenic mice showed significantly increased mean glomerular tuft areas, mean glomerular areas occupied by macrophages, and mean cell numbers per glomerulus compared with wild type controls. All three parameters were decreased in IFN-alpha-treated TSLP transgenic mice, although the differences compared with placebo-treated mice did not reach significance. The changes in glomerular matrix deposition were the same in IFN-alpha-treated and placebo-treated mice. The oral ingestion of IFN-alpha seemed to reduce glomerular macrophage influx, but this did not result in decreased glomerular matrix deposition. The limited positive effect provides experimental support for clinical studies that indicate the beneficial effects of IFN-alpha therapy observed in humans with glomerulonephritis might be attributable to its antiviral effect rather than modulation of intrarenal pathophysiologic pathways. PMID- 11920358 TI - On improving outcomes and quality of dialysis care, and more. PMID- 11920359 TI - A rare cause of nephrotic syndrome. PMID- 11920360 TI - Angiotensin-converting enzyme inhibitors and anemia in chronic kidney disease: a complex interaction. PMID- 11920362 TI - Green urine in a critically ill patient. AB - The development of discolored urine in the critically ill patient, although uncommon, may have many possible causes, with the most likely source related to medication administration. Studies were undertaken in a 39-year-old man who developed dark green urine while in the intensive care unit for neutropenic sepsis. Although the patient had developed prior nonoliguric renal failure stemming from his sepsis, his renal function at the time of presentation of urine discoloration was considered normal. Review of his medications and intravenous infusions suggested the most likely cause was the food dye placed in his enteral tube feedings. Spectrophotometric evaluation of the urine confirmed the presence of Food Dye and Color Blue Number 1 (FD&C Blue No. 1). This case shows that significant gastrointestinal absorption of FD&C Blue No. 1 can occur. FD&C Blue No. 1 should be considered in the differential diagnosis of dark green discolored urine. PMID- 11920363 TI - Rib lesions in a prehistoric Puebloan sample from southwestern Colorado. AB - An unusually high frequency of periosteal lesions of visceral rib surfaces was observed in a small, prehistoric skeletal series from southwestern Colorado. Lesions of this type have been concordant with pulmonary tuberculosis in three studies of human skeletal collections with known cause of death, and in a recent clinical investigation of rib dimensions in living patients with lung disorders. Diseases such as pneumonia and actinomycosis have also been found to cause these lesions, but in much lower frequencies. Archaeological evidence suggests that Puebloan farmers of Sleeping Ute Mountain's southern piedmont, from which the sample is drawn, endured unusually harsh environmental conditions punctuated by severe drought and exacerbated by escalating warfare. It is argued here that these environmental stressors increased susceptibility to an opportunistic respiratory infection reminiscent of tuberculosis, and possibly also some form of pneumonia, resulting in high rates of active disease previously noted only in historic Puebloan peoples. PMID- 11920364 TI - Ancestral loss of the maxillary sinus in Old World monkeys and independent acquisition in Macaca. AB - Cercopithecoid monkeys are unique among primates in that all species (except macaques) lack a maxillary sinus, an unusual condition among eutherian mammals. Although this uncommon distribution of cranial pneumatization was noted previously, the phylogenetic ramifications have not been investigated fully. Recently, character state optimization analysis of computed tomography (CT) data from extant Old World monkeys suggested that the loss of the sinus may have occurred at the origin of the group, unlike previous hypotheses positing only a reduction in size of the structure. To critically evaluate the "early loss" hypothesis, a recently recovered complete cranium of Victoriapithecus macinnesi from Maboko Island, Kenya, was examined by CT to determine the extent of its cranial pneumatization. This taxon is crucial for evaluating character state evolution in Old World monkeys, due to its phylogenetic position, preceding the cercopithecine/colobine split. CT analysis reveals only cancellous bone lateral of the nasal cavity, indicating that Victoriapithecus does not possess a maxillary sinus. Phylogenetic evaluation of the fossil with extant catarrhine taxa strongly supports the early loss of the sinus in cercopithecoids. The results suggest that the maxillary sinus found in the genus Macaca is not homologous with that of other eutherians, which may provide insights into the origin and function (if any) of the paranasal pneumatizations. PMID- 11920365 TI - Deconstructing death in paleodemography. AB - In 1992 in this Journal (Konigsberg and Frankenberg [1992] Am. J. Phys. Anthropol. 89:235-256), we wrote about the use of maximum likelihood methods for the "estimation of age structure in anthropological demography." More specifically, we presented a particular method (the "iterated age-length key") from the fisheries literature and suggested that the method could be used in human and primate demography and paleodemography as well. In our paper (section titled "Some Future Directions"), we spelled out two broad areas that we expected to see develop over the ensuing years. First, we felt that the use of explicit likelihood methods would open up interest in basic estimation issues, such as the calculation of standard errors for demographic estimates and the formulation of tests for whether samples differed in their demographic structure. Second, we felt that the time was ripe for hazards analyses that would incorporate the uncertainty in estimation that follows from using age "indicators" rather than known ages. While some of these developments have occurred during the last decade, few have been reported in the American Journal of Physical Anthropology. In this paper we resolve some issues from our 1992 paper, and attempt to redress this deficit in the literature by reviewing some recent developments in paleodemography over the past decade. PMID- 11920367 TI - Chromosome painting reveals that galagos have highly derived karyotypes. AB - The differences in chromosome number between Otolemur crassicaudatus (2n = 62) and Galago moholi (2n = 38) are dramatic. However, the total number of signals given by hybridizing human chromosome paints to galago metaphases is similar: 42 in O. crassicaudatus and 38 G. moholi. Many human chromosome homologs are found fragmented in each species, and numerous translocations have resulted in chromosomal syntenies or hybridization associations which differ from those found in humans. Only 7 human autosomes showed conserved synteny in O. crassicaudatus, and 9 in G. moholi. Both galago species have numerous associations or syntenies not found in humans: O. crassicaudatus has 11, and G. moholi has 21. The phylogenetic line leading to the last common ancestor of the two galago species accumulated 6 synapomorphic fissions and 5 synapomorphic fusions. Since the divergence of the two galago species, 10 Robertsonian translocations have further transformed the G. moholi karyotype, and 2 fissions have been incorporated into the O. crassicaudatus karyotype. Comparison with other primates, tree shrews, and other mammals shows that both galagos have karyotypes which are a mixture of derived and conserved chromosomes, and neither has a karyotype close to that of the proposed ancestor of all primates. Am J Phys Anthropol 117:319-326, 2002. Published 2002 Wiley-Liss, Inc. PMID- 11920366 TI - DNA decay rate in papyri and human remains from Egyptian archaeological sites. AB - The writing sheets made with strips from the stem (caulis) of papyri (Cyperus papyrus) are one of the most ingenious products of ancient technology. We extracted DNA from samples of modern papyri varying in age from 0-100 years BP and from ancient specimens from Egypt, with an age-span from 1,300-3,200 years BP. The copy number of the plant chloroplast DNA in the sheets was determined using a competitive PCR system designed on the basis of a short (90 bp) tract of the chloroplast's ribulose bisphosphate carboxylase large subunit (rbcL) gene sequence. The results allowed us to establish that the DNA half-life in papyri is about 19-24 years. This means that the last DNA fragments will vanish within no more than 532-672 years from the sheets being manufactured. In a parallel investigation, we checked the archaeological specimens for the presence of residual DNA and determined the extent of racemization of aspartic (Asp) acid in both modern and ancient specimens, as a previous report (Poinar et al. [1996], Science 272:864-866) showed that racemization of aspartic acid and DNA decay are linked. The results confirmed the complete loss of authentic DNA, even in the less ancient (8th century AD) papyri. On the other hand, when the regression for Asp racemization rates in papyri was compared with that for human and animal remains from Egyptian archaeological sites, it proved, quite surprisingly, that the regressions are virtually identical. Our study provides an indirect argument against the reliability of claims about the recovery of authentic DNA from Egyptian mummies and bone remains. PMID- 11920368 TI - Genetic basis of human female pelvic morphology: a twin study. AB - To examine the relative role of genetic and environmental factors on pelvic morphology, data on 60 pairs of female twins (30 monozygotic (MZ) and 30 dizygotic (DZ)) were analyzed. Fourteen pelvic measurements were normally distributed, and two were not. Association of twin type with the mean value of a trait was found in only 1 out of 8 traits. Heterogeneity of variance between zygosities was observed in 4 pelvic traits (50%), invalidating within-pair estimates of genetic variance for these traits. Evidence of stronger environmental covariance for MZ than DZ twins was observed for only one trait (sitting height iliocristale). A significant genetic component of variation was observed for age at menarche and in the pelvic area. In instances where inequality of variances between zygosities was demonstrated, total among-pair and within-pair mean squares were larger for dizygotic than for monozygotic twins. This is interpreted as evidence of greater environmental influence between zygosities. Environmental modification was not of the same magnitude in various pelvic traits. Bitrochanteric breadth had the highest magnitude of cultural heritability, indicating that cultural factors played an important role in determining hip breadth. PMID- 11920369 TI - Testing isonymy with paternal and maternal lineages in the early Quebec population: the impact of polyphyletism and demographic differentials. AB - Isonymy is an ingenious and useful approach to studying kinship in human populations. However, it relies on assumptions that are difficult to verify. In this study, we provided a way to assess, in the early Quebec population, the impact of factors such as polyphyletism, unbalanced sex-ratio among founders, and age differentials between spouses. All data were taken from the Population Register of Early Quebec, which contains births, marriages, and deaths (>712,000) recorded in parish registers from the beginning of colonization (in 1608) to 1800. More specifically, using the 70,869 marriages recorded during that period, we compared kinship estimates given by genealogies, surnames, and paternal and maternal lineages. We also calculated a fifth coefficient of kinship by combining paternal and maternal lineage, thus providing a new way to test the isonymy method. The results show a good agreement between genealogical and isonymous estimates. However, this good correspondence is due to counterbalancing biases. Some of the implications of our results are discussed in the context of colonial America. PMID- 11920370 TI - New perspectives on taste and primate evolution: the dichotomy in gustatory coding for perception of beneficent versus noxious substances as supported by correlations among human thresholds. AB - In various environments where primates are presently observed, as well as in forests and savannas which have been inhabited by australopithecines and early hominids, there are (or there have been presumably) categories of substances eliciting taste signals associated with stereotyped responses. Such is the case for various soluble sugars of fruits and nectars, attracting consumers, and for several plant compounds in which bitter or strongly astringent properties have a repulsive effect. The occurrence of such classes of tasty substances among natural products appears to be related to the evolutionary trends that shaped primate sensory perception (for detecting either beneficent or potentially noxious substances) in the context of a long history of coevolution between animals and plants. Here, we present original psychophysical data on humans (412 individuals aged 17-59 years) as an analogy with which to test recent evidence from electrophysiology in nonhuman primates (Hellekant et al. [1997] J. Neurophysiol. 77:978-993; Danilova et al. [1998] Ann. N.Y. Acad. Sci. 855:160 164) that taste fibers can be grouped into clusters of "best-responding fibers" with two more specific clusters, one for sugars and one for quinine and tannins. The collinearity found between human taste responses (recognition thresholds) for fructose and sucrose, as well as for quinine and tannins, is presented and discussed as another evidence of the two-direction evolutionary trend determining taste sensitivity. Salt perception appears to be totally independent of these trends. Accordingly, the appreciation of a salty taste seems to be a recent culturally learned response, and not a primary taste perception. The very existence of primary tastes is discussed in the context of evolutionary trends, past and present. PMID- 11920371 TI - Scale issues in the study of primate foraging: red colobus of Kibale National Park. AB - Diet data have been used to address a number of theoretical issues. We often calculate the proportion of time spent eating different foods (e.g., fruits, leaves) to place species into dietary categories and contrast morphological or behavioral traits among categories. Yet we have little understanding of how flexible species can be in terms of the plant parts and species consumed. To address this issue, we analyzed data on the diets of red colobus monkeys (Procolobus badius) from Kibale National Park, Uganda, to evaluate temporal and spatial variability in the plant parts and species eaten. After considering observer differences and sampling issues, we evaluated how different a group's diet could be if samples were taken in different years. We found that the diet of the same groups showed significant, consistent changes over a 4-year period. For example, the time spent feeding on leaves increased from 56% in 1994 to 76% in 1998. The plant parts and species eaten by eight groups inhabiting different types of forest (e.g., pristine, logged, riverine) varied among groups. The largest interdemic difference was seen in the use of young leaves (38%). Dietary differences were also found between groups with overlapping home ranges (41-49% overlap). Different subspecies of Procolobus badius also varied in diet; however, this variation was often not of the magnitude documented within Kibale for the same population. The fact that diet can vary considerably over small spatial and short temporal scales within the same species raises the intriguing question as to what level of interspecific difference is biologically significant for addressing particular questions. We conclude that behavioral flexibility blurs our traditional stereotypic assessment of primates; a study of one group that occupies a specific habitat at one point in time may not adequately represent the species. PMID- 11920372 TI - Growth and development of adolescent girls from the Segou Region of Mali (West Africa). AB - In Mali, adolescent girls face many physical stresses which, with a history of poor nutrition, can result in delayed growth and development. Despite the fact that adolescent girls encounter nutritional demands, most research in Mali has focused only on young children. This study examines anthropometric and reproductive data on 1,056 adolescent girls (aged 10-17 years) from the Segou Region of Mali. When compared to the reference population, the Malian girls exhibited poorer indicators of growth and development. Z-scores for height-for age and weight-for-age were below 0 at all ages. Urban girls had better indicators of growth than rural girls. The body composition data show that the Malian girls have lower body fat than reference girls. Menarche was delayed by about 1.5 years compared to girls from the United States and Europe. Delayed growth and development may be due to many factors, including a history of undernutrition, poor socioeconomic status, and increased energy demands. PMID- 11920381 TI - Belief vs. scientific observation: the curious story of the precardiac mesoderm. PMID- 11920382 TI - Daily morphological variations in the viscacha (Lagostomus maximus maximus) retina. Probable local modulatory action of melatonin. AB - Given that the local melatonin levels exhibit rhythmic daily changes in the retina of the viscacha, we considered it important to study the likely daily variations in morphology and specific 2-[(125)I]-iodomelatonin binding in retinas from this rodent and to correlate these putative changes with local indole levels. Adult animals of both sexes were captured in their habitat and were kept under a natural photoperiod. For light and electron microscopic studies the viscachas were sacrificed by decapitation at 08:00, 16:00, and 24:00 hr. A computer-assisted image analysis system was used to measure the thickness of the complete retina, the photoreceptor layer, the rod outer and inner segments, and the outer nuclear layer. The daily variation in 2-[(125)I]-iodomelatonin binding sites was followed during a 24-hr light-dark cycle, the animals being sacrificed at six time points. The parameters studied showed significant variations throughout the 24-hr period. Maximal specific binding, lysosomal content in the pigment epithelium, and photoreceptor layer outer segment thicknesses were observed at 24:00 hr. Close contact between photoreceptor membranes and microvilli of the pigment epithelium was observed at 08:00 and 16:00 hr. Moreover, the minimal outer segment thickness at 16:00 hr was accompanied by a scarcity of dense bodies, such as lysosomes, a maximum dispersion of melanin pigment granules, and a minimum density of radioligand binding sites. Therefore, in the retina of the viscacha, we suggest that the interaction between melatonin and specific sites could be one of the factors or causes that participate in the regulation of the daily morphological changes observed in viscacha. PMID- 11920384 TI - Osteoclast induction in periodontal tissue during experimental movement of incisors in osteoprotegerin-deficient mice. AB - Osteoprotegerin (OPG) is a novel secreted member of the tumor necrosis factor (TNF) receptor superfamily that negatively regulates osteoclastogenesis. The receptor activator of the NFKB ligand (RANKL) is one of the key regulatory molecules in osteoclast formation and binds to OPG. In this study, it was suggested that OPG and RANKL are involved in alveolar bone remodeling during orthodontic tooth movement. We examined RANKL localization and osteoclast induction in periodontal tissues during experimental movement of incisors in OPG deficient mice. To produce orthodontic force, an elastic band was inserted between the upper right and left incisors for 2 or 5 days, and the dissected maxillae were examined for cytochemical and immunocytochemical localization of tartrate-resistant acid phosphatase (TRAP), vacuolar-type H(+)-ATPase, and RANKL. Compared to wild-type OPG (+/+) littermates, TRAP-positive multinucleated cells were markedly induced in the periodontal ligament (PDL) on the compressed side and in the adjacent alveolar bone of OPG-deficient mice. These multinucleated cells exhibited intense vacuolar-type H(+)-ATPase along the ruffled border membranes. Because of accelerated osteoclastic resorption in OPG-deficient mice, alveolar bone was severely destroyed and partially perforated at 2 and 5 days after force application. In both wild-type and OPG-deficient mice, RANKL expression became stronger at 2 and 5 days after force application than before force application. There was no apparent difference in intensity of RANKL expression between OPG (+/+) littermates and OPG-deficient mice. In both wild type and OPG-deficient mice, expression of RANKL protein was detected in osteoblasts, fibroblasts, and osteoclasts mostly located in resorption lacunae. These results suggest that during orthodontic tooth movement, RANKL and OPG in the periodontal tissues are important determinants regulating balanced alveolar bone resorption. PMID- 11920383 TI - A comparative study of gut-associated lymphoid tissue in calf and chicken. AB - The calf contains two types of Peyer's patches (PPs): jejunal and ileal. The ileal PP has been thought to be equivalent to the bursa of Fabricius (BF) as a central lymphoid organ. The morphologies of ileal and jejunal PPs in the calf were compared with those of the BF and the caecal tonsil (CT) in the chicken. Immunoglobulin G-positive (IgG(+)) cells appear in the follicles of them all and exhibited a dendritic appearance after birth. We investigated whether the IgG in these follicles was produced in situ. IgG-producing cells were detected in the follicular medullas of the jejunal PP and the CT, but not in those of the ileal PP and the BF. CD4(+) cells were distributed in the follicular medullas of the jejunal PP and the CT, but not in those of the ileal PP and the BF. The data suggest that Ig class switching occurs in both jejunal PP follicles and CT follicles, but does not occur in either the ileal PP follicles or the bursal follicles. Because CD4(+) T cells would be prerequisite for Ig class switching in these follicles, IgG(+) cells of the follicular medullas in the ileal PP and the BF would trap immune complexes from the gut lumen. The primary B-cell repertoire might be selected by gut-derived antigens in the ileal PP and the BF before seeding the periphery. PMID- 11920385 TI - GABA-, glycine-, and glutamate-immunoreactive bouton profiles in apposition to neurons of the central cervical nucleus in the rat. AB - The neurons of the central cervical nucleus (CCN) convey information about the position and movements of the head, and receive excitatory input from dorsal neck muscles and the labyrinth. Both of these afferent sources form glutamatergic synaptic contacts with CCN neurons. However, these sensory afferent sources can also inhibit CCN neurons. To further elucidate the synaptic organization, we made an electron microscopic investigation, identifying and evaluating the relative frequency of bouton profiles containing the inhibitory transmitters GABA and glycine in apposition to identified CCN neurons. In addition, labeling for glutamate was performed. The identification of the CCN neurons was made possible by injections of retrograde tracer substances into the cerebellum. These substances were made visible by preembedding immunocytochemistry or postembedding immunogold staining. Such staining was also used to detect the three amino acids that were found in boutons apposed to the identified neurons (cf. Ornung et al., J. Comp. Neurol. 1996;365:413-426; Linda et al., J. Comp. Neurol. 2000;425:10 23). Due to the relatively poor transport of the tracer substances into dendrites of the CCN neurons, the analysis was restricted to the cell body and included bouton profiles in direct apposition to the soma membrane. Data from 10 CCN neurons revealed that about 50% of the apposing bouton profiles were immunoreactive for GABA, and about 34% for glycine. In four neurons, the degree of colocalization of GABA and glycine was determined to be close to 30%. Thus, the vast majority of glycine-labeled profiles also contained GABA, while a considerable fraction of the profiles were immunoreactive for only GABA. The values for glycine immunoreactive bouton profiles presented here may represent somewhat low estimates, depending on the method used. Data from four neurons showed that about 18% of the profiles were labeled for glutamate. The large fraction of purely GABA immunoreactive profiles, or at least a substantial group of them, is suggestive of their derivation from axons descending from the brainstem. PMID- 11920386 TI - Cell kinetic study of the endometrium by nonisotopic in situ hybridization for histone H3 messenger RNA and immunohistochemistry for Ki-67 and for estrogen and progesterone receptors. AB - We investigated the cell kinetics of the endometrium in hysterectomy specimens taken for leiomyoma from 22 women with regular ovulatory menstrual cycles. Formalin-fixed, paraffin-embedded tissue sections were examined for proliferating activity using histone H3 messenger RNA in situ hybridization (H3 mRNA-ISH) and immunostaining for the Ki-67 antigen. The relationship of the proliferative activity of endometrial cells to the immunohistochemical expression of the estrogen receptor (ER) and the progesterone receptor (PR) was also examined. During the menstrual cycle, H3 mRNA expression was observed in both the epithelial cells and the stromal cells of the endometrium. In the functional layer, the labeling indices for H3 mRNA (H3 mRNA-LIs) in the epithelial cells peaked in the late proliferative phase, decreased sharply in the early secretory phase, and remained unchanged thereafter. On the other hand, H3 mRNA-LIs of stromal cells displayed two peaks: one in the midproliferative phase and the other in the late secretory phase, the former peak being the greater. In the basal layer, epithelial cells and stromal cells showed low H3 mRNA-LIs and no significant variation throughout the menstrual cycle. The H3 mRNA-LIs correlated well with the Ki-67-LIs and were lower than the corresponding Ki-67-LIs. The regression coefficient (H3 mRNA-LIs against the Ki-67-LIs) was 0.33 for epithelial cells and 0.49 for stromal cells, suggesting that the cell cycle time was longer for epithelial cells than for stromal cells. The proliferative activity of endometrial cells showed close relationships with the expressions of ER and PR in the endometrium. When used in combination with other proliferative markers in paraffin-embedded tissue sections, H3 mRNA-ISH could open broader perspectives on the cell kinetics of the endometrium. PMID- 11920387 TI - Horizontally oriented clusters of multiple chondrons in the superficial zone of ankle, but not knee articular cartilage. AB - Osteoarthritis is a progressive disease that is initiated at the surface of articular cartilage and proceeds to destroy the entire depth of the cartilage. The prevalence of osteoarthritis varies in different joints; e.g., the ankle joint has a very low prevalence of the disease compared to the knee joint. To better understand any inherent differences between the articular cartilage of the ankle and that of the knee that would account for the difference in occurrence of osteoarthritis, studies were undertaken to examine differences between the superficial zones in these two joint cartilages obtained from human donors. Chondrocytes in the superficial zones of the normal ankle (talocrural) and the normal knee (tibiofemoral) joints were identified with a monoclonal antibody specific for the superficial zone protein (SZP). When the chondrocytes from both joints were compared in serial horizontal sections, the chondrocytes in the superficial zone of the knee cartilage were seen either as isolated single cells or as doublets. However, the chondrocytes within the superficial zone of normal ankle cartilage were arranged in planar clusters containing multiple chondrons composed of 2-13 cells. There were no detectable differences in the chondrocyte clusters in the superficial zone of the ankle with respect to age, gender, or site on the cartilage surface. Adjacent to a lesion in an ankle joint with degenerative changes, the clusters were larger, containing up to 22 chondrocytes. This is the first report documenting the presence of multiple chondrons in the superficial zone of normal human adult articular cartilage. PMID- 11920388 TI - Missing Omo L338y-6 occipital-marginal sinus drainage pattern: ground sectioning, computer tomography scanning, and the original fossil fail to show it. AB - The Omo L338y-6 occipital region has been recently studied by White and Falk (1999), who claim that it shows a readily identifiable enlarged left occipital marginal sinus (O/M). These observations are contrary to the direct observations of previous investigators (Rak and Howell, 1978; Kimbel, 1984; Holloway, 1981; Holloway, 1988). White and Falk (1999) further argue that the presence of this enlarged O/M strongly suggests that the Omo L338y-6 hominid was indeed a "robust" Australopithecus. We used direct sectioning and CT scanning to analyze magnified sections of a high-quality first-generation cast of the newly cleaned original fossil. These methods fail to show any evidence of a morphological landmark that can be interpreted as an enlarged O/M, either as an eminence or a sulcus. In contrast, the same techniques used with both SK 1585 and OH5 ("robust" Australopithecus with an enlarged O/M) show extremely visible and palpable enlarged O/M's. Examination of the original Omo fossil confirms that it lacks an O/M. This evidence clearly shows that an enlarged O/M cannot be identified on either the original fossil or a first-generation cast, although this does not rule out the possibility that the Omo L338y-6 hominid was a "robust" Australopithecus. We believe that the differences between observers regarding this feature are most probably due to displacement caused by a crack and the different source materials employed, i.e., the difference between a first generation cast of the original fossil and a third- or fourth-generation cast of the endocast made two decades ago. PMID- 11920389 TI - The best of times, the worst of times: rheumatology 2001. PMID- 11920390 TI - Targeting interleukin-1 in the treatment of rheumatoid arthritis. PMID- 11920391 TI - The changing face of rheumatoid arthritis: why the decline in incidence? PMID- 11920392 TI - The search for rheumatoid arthritis susceptibility genes: a call for global collaboration. PMID- 11920393 TI - Hepatitis C virus infection and vasculitis: implications of antiviral and immunosuppressive therapies. PMID- 11920394 TI - Peroxisome proliferator-activated receptor gamma agonists: potential use for treating chronic inflammatory diseases. PMID- 11920395 TI - The cutting edge of spondylarthropathy research in the millennium. PMID- 11920396 TI - Treatment of rheumatoid arthritis with anakinra, a recombinant human interleukin 1 receptor antagonist, in combination with methotrexate: results of a twenty-four week, multicenter, randomized, double-blind, placebo-controlled trial. AB - OBJECTIVE: To evaluate the efficacy and safety of anakinra in combination with methotrexate (MTX) in patients with active rheumatoid arthritis (RA). METHODS: Patients with moderate-to-severe active RA who were receiving MTX for 6 consecutive months, with stable doses for > or = 3 months (those with disease duration of >6 months but <12 years) were randomized into 6 groups: placebo or 0.04, 0.1, 0.4, 1.0, or 2.0 mg/kg of anakinra administered in a single, daily, subcutaneous injection. The primary efficacy end point was the proportion of subjects who met the American College of Rheumatology 20% improvement criteria (attained an ACR20 response) at week 12. RESULTS: A total of 419 patients were randomized in the study. Patient demographics and disease status were similar in the 6 treatment groups. The ACR20 responses at week 12 in the 5 active treatment plus MTX groups demonstrated a statistically significant (P = 0.001) dose response relationship compared with the ACR20 response in the placebo plus MTX group. The ACR20 response rate in the anakinra 1.0-mg/kg (46%; P = 0.001) and 2.0 mg/kg (38%; P = 0.007) dose groups was significantly greater than that in the placebo group (19%). The ACR20 responses at 24 weeks were consistent with those at 12 weeks. Similar improvements in anakinra-treated subjects were noted in individual ACR components, erythrocyte sedimentation rate, onset of ACR20 response, sustainability of ACR20 response, and magnitude of ACR response. Anakinra was safe and well tolerated. Injection site reaction was the most frequently noted adverse event, and this led to premature study withdrawal in 7% (1.0-mg/kg group) to 10% (2.0-mg/kg group) of patients receiving higher doses. CONCLUSION: In patients with persistently active RA, the combination of anakinra and MTX was safe and well tolerated and provided significantly greater clinical benefit than MTX alone. PMID- 11920397 TI - Trends in incidence and mortality in rheumatoid arthritis in Rochester, Minnesota, over a forty-year period. AB - OBJECTIVE: To determine time trends in the epidemiology of rheumatoid arthritis (RA) in a population-based cohort. METHODS: An inception cohort of residents of Rochester, Minnesota > or = 18 years of age who first fulfilled the American College of Rheumatology 1987 criteria between January 1, 1955 and December 31, 1994 (applied retrospectively, as appropriate) was assembled and followed up until January 1, 2000. Incidence rates were estimated and were age- and sex adjusted to the 1990 white population of the US. A birth cohort analysis was performed, and survival rates over time were examined. RESULTS: The incidence cohort comprised 609 patients, 445 (73.1%) of whom were female and 164 (26.9%) were male, with a mean age at incidence of 58.0 years. The overall age- and sex adjusted annual incidence of RA among Rochester, Minnesota, residents > or = 18 years of age was 44.6/100,000 population (95% confidence interval 41.0-48.2). While the incidence rate fell progressively over the 4 decades of study, from 61.2/100,000 in 1955-1964, to 32.7/100,000 in 1985-1994, there were indications of cyclical trends over time. Birth cohort analysis showed diminishing incidence rates through successive cohorts following a peak in the 1880-1890 cohorts. Incidence rates increased with age until age 85, but peaked earlier in women than in men. The survival rate in RA patients was significantly lower than the expected rate in the general population (P < 0.001), and no improvement was noted over time. CONCLUSION: The secular trends demonstrated in this study population, including the progressive decline in the incidence of RA over the last 40 years, suggest that an environmental factor may play a role in the etiology of RA. PMID- 11920398 TI - Whole-genome linkage analysis of rheumatoid arthritis susceptibility loci in 252 affected sibling pairs in the United Kingdom. AB - OBJECTIVE: To undertake a systematic whole-genome screen to identify regions exhibiting genetic linkage to rheumatoid arthritis (RA). METHODS: Two hundred fifty-two RA-affected sibling pairs from 182 UK families were genotyped using 365 highly informative microsatellite markers. Microsatellite genotyping was performed using fluorescent polymerase chain reaction primers and semiautomated DNA sequencing technology. Linkage analysis was undertaken using MAPMAKER/SIBS for single-point and multipoint analysis. RESULTS: Significant linkage (maximum logarithm of odds score 4.7 [P = 0.000003] at marker D6S276, 1 cM from HLA-DRB1) was identified around the major histocompatibility complex (MHC) region on chromosome 6. Suggestive linkage (P < 7.4 x 10(-4)) was identified on chromosome 6q by single- and multipoint analysis. Ten other sites of nominal linkage (P < 0.05) were identified on chromosomes 3p, 4q, 7p, 2 regions of 10q, 2 regions of 14q, 16p, 21q, and Xq by single-point analysis and on 3 sites (1q, 14q, and 14q) by multipoint analysis. CONCLUSION: Linkage to the MHC region was confirmed. Eleven non-HLA regions demonstrated evidence of suggestive or nominal linkage, but none reached the genome-wide threshold for significant linkage (P = 2.2 x 10( 5)). Results of previous genome screens have suggested that 6 of these regions may be involved in RA susceptibility. PMID- 11920399 TI - Smoking and disease severity in rheumatoid arthritis: association with polymorphism at the glutathione S-transferase M1 locus. AB - OBJECTIVE: To determine whether the relationship between smoking and disease severity in women with rheumatoid arthritis (RA) is associated with polymorphism at the glutathione S-transferase (GST) M1 locus. METHODS: Genotyping for GSTM1 was carried out using polymerase chain reaction methodology on 164 women with established RA. Smoking history was obtained on each patient. Radiographic damage was measured by the Larsen score, and functional outcome was assessed by the Health Assessment Questionnaire (HAQ). Data were analyzed by multiple regression analyses, with correction for age and disease duration. RESULTS: Ever having smoked was associated with a worse radiographic and functional outcome than was never having smoked. Both past and current smoking were associated with increased disease severity. Stratification by GSTM1 status revealed that polymorphism at this locus affected the relationship between smoking and disease outcome measures. Patients who lacked the GSTM1 gene and had ever smoked had significantly higher Larsen and HAQ scores than did those who lacked the gene and had never smoked. Radiographic outcome in these patients was worse than that in patients who had the GSTM1 gene and who had smoked. The associations were not affected by correction for socioeconomic status. Rheumatoid factor (RF) production was found to be associated with smoking in only the GSTM1-null patients. CONCLUSION: Our data suggest that disease outcome in female RA patients with a history of smoking is significantly worse than in those who have never smoked. Smoking was associated with the most severe disease in patients who carried the GSTM1-null polymorphism. This association may be due in part to a relationship between the GSTM1 polymorphism and RF production in smokers. PMID- 11920400 TI - The value of contrast-enhanced color Doppler ultrasound in the detection of vascularization of finger joints in patients with rheumatoid arthritis. AB - OBJECTIVE: A prospective study was performed to assess the usefulness of contrast enhanced color Doppler ultrasound (CDUS) in the evaluation of intraarticular vascularization of finger joints in patients with rheumatoid arthritis (RA). METHODS: We investigated 198 finger joints in 46 patients with RA, and 80 finger joints in 10 healthy volunteers. Joints with varying levels of clinical activity of inflammation were classified as being active, moderately active, or inactive. CDUS was performed with a high-frequency multi-D linear array transducer. A microbubble-based ultrasound (US) contrast agent (Levovist; Schering, Berlin, Germany) was intravenously infused. Doppler findings were rated on the basis of both unenhanced and contrast-enhanced CDUS images. RESULTS: Healthy joints showed no intraarticular vascularization on either unenhanced or contrast-enhanced CDUS. Unenhanced CDUS detected intraarticular vascularization in 7 (8%) of 83 inactive joints, in 31 (52%) of 60 moderately active joints, and in 32 (58%) of 55 active joints. Contrast-enhanced CDUS detected intraarticular vascularization in 41 (49%) of 83 joints with inactive RA, in 59 (98%) of 60 joints with moderately active RA, and in all 55 joints with active RA. Detection of intraarticular vascularization was improved by administration of the microbubble-based US contrast agent (P < 0.001). Contrast-enhanced CDUS demonstrated differences in intraarticular vascularization between joints with inactive RA and those with active RA (P < 0.001), between joints with inactive RA and those with moderately active RA (P < 0.001), and between joints with moderately active RA and those with active RA (P < 0.001). CONCLUSION: The use of a microbubble-based US contrast agent significantly improved the detection of intraarticular vascularization in the finger joints of patients with RA. This technique seems to be a useful adjunct in the assessment of disease activity. PMID- 11920401 TI - Inadequately low serum levels of steroid hormones in relation to interleukin-6 and tumor necrosis factor in untreated patients with early rheumatoid arthritis and reactive arthritis. AB - OBJECTIVE: To compare levels of steroid hormones in relation to cytokines and to study levels of cortisol or dehydroepiandrosterone (DHEA) in relation to other adrenal hormones in untreated patients with early rheumatoid arthritis (RA) and reactive arthritis (ReA) compared with healthy controls. METHODS: In a retrospective study with 34 RA patients, 46 ReA patients, and 112 healthy subjects, we measured serum levels of interleukin-6 (IL-6), tumor necrosis factor (TNF), adrenocorticotropic hormone (ACTH), cortisol, 17-hydroxyprogesterone (17 OH-progesterone), androstenedione (ASD), DHEA, and DHEA sulfate (DHEAS). RESULTS: RA patients had higher serum levels of IL-6, TNF, cortisol, and DHEA compared with ReA patients and healthy subjects, but no difference was noticed with respect to ACTH and DHEAS. However, in RA and ReA patients compared with healthy subjects, levels of ACTH, cortisol, ASD, DHEAS, and 17-OH-progesterone were markedly lower in relation to levels of IL-6 and TNF. Furthermore, the number of swollen joints correlated inversely with the ratio of serum cortisol to serum IL 6 in RA (R(Rank) = -0.582, P = 0.001) and, to a lesser extent, in ReA (R(Rank) = 0.417, P = 0.011). In RA patients, the mean grip strength of both hands was positively correlated with the ratio of serum cortisol to serum IL-6 (R(Rank) = 0.472, P = 0.010). Furthermore, in these untreated patients with RA and ReA, there was a relative decrease in the secretion of 17-OH-progesterone, ASD, and DHEAS in relation to DHEA and cortisol. This indicates a relative predominance of the nonsulfated DHEA and cortisol in relation to all other measured adrenal steroid hormones in the early stages of these inflammatory diseases. CONCLUSION: This study indicates that levels of ACTH and cortisol are relatively low in relation to levels of IL-6 and TNF in untreated patients with early RA and ReA compared with healthy subjects. The study further demonstrates that there is a relative increase of DHEA and cortisol in relation to other adrenal hormones, such as DHEAS. This study emphasizes that adrenal steroid secretion is inadequately low in relation to inflammation. Although changes in hormone levels are similar in RA and ReA, alteration of steroidogenesis is more pronounced in RA patients than in ReA patients. PMID- 11920402 TI - Comparison of cathepsins K and S expression within the rheumatoid and osteoarthritic synovium. AB - OBJECTIVE: To determine and compare the expression of cathepsins K and S proteins in joints with rheumatoid arthritis (RA) and osteoarthritis (OA) and to determine the effect of interleukin-1 beta (IL-1 beta) and tumor necrosis factor alpha (TNF alpha) on the expression of cathepsin K in fibroblast-like synoviocytes. METHOD: Expression and localization of cathepsins K and S were determined by immunohistochemistry in the synovium of 10 RA- and 8 OA-affected joints. Northern and Western blot analyses were performed to analyze cathepsin K and S expression in primary fibroblast-like synoviocyte cultures from RA and OA patients. The effect of IL-1 beta and TNF alpha on the expression and secretion of cathepsin K in primary cultures of synoviocytes was determined by real-time polymerase chain reaction and Western blot analysis. Staining of in situ activity was used to identify active cathepsin K enzyme in primary synovial fibroblast cultures. RESULTS: Cathepsin K and S protein expression was identified in the synovium from patients with RA and OA. Cathepsin K protein was localized in synovial fibroblasts, stromal multinucleated giant cells, and, to a lesser degree, in CD68+ macrophage-like synoviocytes. Of note is the expression of cathepsin K in synovial fibroblasts and mononuclear macrophage-like cells at sites of cartilage erosion in RA and in interdigitating cells of lymphocyte-rich areas. In contrast, cathepsin S expression was restricted to CD68+ macrophage-like synoviocytes, interdigitating cells, and endothelial cells of blood vessels. Cathepsin K protein expression in the interstitial areas and perivascular regions of RA derived synovial specimens was 2-5 times higher than in OA samples (P < 0.001), whereas the expression of cathepsin S did not significantly differ in these diseases. Cathepsin K expression levels in normal synovium were low and restricted to fibroblast-like cells. Of note, cathepsin K also was expressed in repairing fibrocartilage in 1 OA specimen. Primary cell cultures of RA- and OA derived synovial fibroblasts expressed comparable amounts of cathepsin K at the transcript and protein levels. Both cell cultures secreted mature cathepsin K as well as procathepsin K, and expressed active cathepsin K in cytosolic vesicles. In contrast, neither RA- nor OA-derived fibroblasts expressed detectable levels of cathepsin S. IL-1 beta and TNF alpha stimulated the transcript (7-8-fold) and protein expression (2-fold) of cathepsin K (P < 0.05) in primary synovial fibroblast cultures, without differences in expression between RA- and OA-derived synovial fibroblasts. CONCLUSION: The presence of cathepsin K polypeptide in synovial fibroblasts and macrophage-like cells in normal, OA, and RA synovia suggests a constitutive expression of this protease and a role in synovial remodeling. The comparable increase in cathepsin K expression after stimulation of RA- and OA-derived synovial fibroblasts with IL-1 beta and TNF alpha further suggests that the expression of cathepsin K is independent of cellular alterations leading to the invasive phenotype of RA-synovial fibroblasts. However, the overexpression of cathepsin K in RA synovia due to an increase in the number of cathepsin K-expressing cells identifies this enzyme as a candidate protease for the pathologic degradation of articular cartilage. Cathepsin S expression in macrophage-like synoviocytes suggests dual activity in antigen presentation and matrix degradation in the inflamed synovia. PMID- 11920403 TI - Risk factors for total hip replacement due to primary osteoarthritis: a cohort study in 50,034 persons. AB - OBJECTIVE: To identify prospectively the possible risk factors for total hip replacement (THR) due to primary osteoarthritis in a large cohort. METHODS: Data from a cardiovascular screening were matched with 9 years of national data on THR. Mean age at the start of followup was 54.9 years, and the 50,034 participants were followed up for an average of 9 years. During followup, 672 persons had a first THR due to primary osteoarthritis. RESULTS: We found dose response associations between body mass index (BMI), body weight, and the level of physical activity at work and THR for primary osteoarthritis. The highest versus the lowest quarter of BMI had a relative risk of 2.0 (95% confidence interval [95% CI] 1.4-2.9) among men and 3.0 (95% CI 2.1-4.1) among women. The highest versus the lowest quarter of body weight had a relative risk of 2.1 (95% CI 1.4-3.2) among men and 3.4 (95% CI 2.4-4.9) among women. Intensive versus sedentary physical activity at work had a relative risk of 2.1 (95% CI 1.5-3.0) among men and 2.1 (95% CI 1.3-3.3) among women. No association was found between physical activity in leisure and THR for primary osteoarthritis. CONCLUSION: Intensive physical activity at work and a high BMI each contribute significantly to the overall risk of undergoing THR due to primary osteoarthritis. Lowering the exposure to these risk factors may substantially reduce the need for hip replacement. PMID- 11920404 TI - Increased life span of human osteoarthritic chondrocytes by exogenous expression of telomerase. AB - OBJECTIVE: To extend the life span of human osteoarthritic (OA) articular chondrocytes by introduction of the catalytic component of human telomerase while preserving the chondrocyte-specific phenotype. METHODS: Human articular chondrocytes were isolated from the femoral head and tibial plateau of patients undergoing knee joint replacement for OA. The chondrocytes were cultured as monolayers and infected with a retroviral telomerase expression construct followed by selection with G418 for 10-14 days. Telomeric-repeat amplification protocol assays and telomere terminal restriction fragment length assays were performed on pools of transduced cells in order to measure telomerase activity and telomere length. Growth kinetics and population doubling capacity were assessed by passaging the cells in monolayer culture. Redifferentiation of the monolayer chondrocyte cultures was induced by transfer to suspension culture on poly-(2-hydroxyethyl-methacrylate) (polyHEMA)-coated dishes. Induction of the chondrocyte-specific phenotype was monitored by analysis of gene expression utilizing reverse transcription-polymerase chain reaction. RESULTS: OA chondrocytes isolated from 3 different donors (ages 41, 69, and 75 years) were transduced with a retroviral construct expressing telomerase. After selection, pooled populations of cells from all donors and a clonal cell line from 1 donor expressed telomerase activity and exhibited lengthening of telomeres. Chondrocytes expressing telomerase showed an increase of 5-9 population doublings over 234 days of culture in monolayer. The telomerase-transduced cells recovered a chondrocyte-specific gene expression pattern following culture on polyHEMA coated dishes. CONCLUSION: The exogenous expression of telomerase may represent a way to expand human OA chondrocytes while allowing maintenance of the chondrocyte specific phenotype. These cells have the potential to be used for restoration of the articular cartilage defects occurring in this disease. PMID- 11920405 TI - Inhibition of insulin-like growth factor binding protein 5 proteolysis in articular cartilage and joint fluid results in enhanced concentrations of insulin like growth factor 1 and is associated with improved osteoarthritis. AB - OBJECTIVE: The complement component C1s is present in dog joint fluid in an activated state. Since C1s degrades insulin-like growth factor binding protein 5 (IGFBP-5), we undertook to determine whether inhibiting C1s in joint fluid would result in an increase in the amount of intact IGFBP-5 and IGF-1 in cartilage and joint fluid, and whether C1s inhibition would be associated with a reduction in cartilage destruction during the development of osteoarthritis (OA). METHODS: Twenty-two dogs were randomized to 3 treatment groups. All dogs underwent anterior cruciate ligament transection and were exercised. Dogs received 1 of 3 treatments: buffer alone (controls; n = 6); PB-145, a peptide derived from the sequence of antithrombin III (n = 9); and pentosan polysulfate (PPS; n = 7). PB 145 or saline was injected into the joint space 3 times per week for 3 weeks. PPS was injected intramuscularly weekly for 3 weeks. RESULTS: Joint histology showed preservation of chondrocytes and a smooth joint surface in the animals treated with PB-145 and PPS. Mankin scoring showed statistically significant reductions in joint destruction with PB-145 and PPS treatments (P < 0.01) compared with buffer control. Mean active collagenase concentrations were decreased by these two treatments. Immunoblotting of joint fluid showed that both treatments increased concentrations of intact IGFBP-5. Direct analysis of IGFBP-3 and IGFBP 5 protease activity showed that IGFBP-5 was degraded more rapidly and that PB-145 and PPS inhibited the degradation of both proteins. Total IGF-1 concentrations in joint fluid were increased 5.6-5.8-fold by these two treatments. Analysis showed that C1s was being activated in joint fluid and that its activation was inhibited by the addition of PB-145 or PPS. CONCLUSION: The findings suggest that direct inhibition of the serine protease C1s results in increased concentrations of intact IGFBP-5 and that proteolysis of IGFBP-3 is also inhibited, probably by the inhibition of some other protease. This increase in concentrations of intact IGFBP-3 and IGFBP-5 leads to an increase in IGF-1 which is associated with an improvement in joint architecture during the development of OA. PMID- 11920406 TI - Reduced chondrogenic and adipogenic activity of mesenchymal stem cells from patients with advanced osteoarthritis. AB - OBJECTIVE: Mesenchymal stem cells (MSCs) are resident in the bone marrow throughout normal adult life and have the capacity to differentiate along a number of connective tissue pathways, among them bone, cartilage, and fat. To determine whether functionally normal MSC populations may be isolated from patients with advanced osteoarthritis (OA), we have compared cells from patients undergoing joint replacement with cells from normal donors. Cell populations were compared in terms of yield, proliferation, and capacity to differentiate. METHODS: MSCs were prepared from bone marrow aspirates obtained from the iliac crest or from the tibia/femur during joint surgery. In vitro chondrogenic activity was measured as glycosaminoglycan and type II collagen deposition in pellet cultures. Adipogenic activity was measured as the accumulation of Nile Red O-positive lipid vacuoles, and osteogenic activity was measured as calcium deposition and by von Kossa staining. RESULTS: Patient-derived MSCs formed colonies in primary culture that were characteristically spindle-shaped with normal morphology. The primary cell yield in 36 of 38 cell cultures from OA donors fell within the range found in cultures from normal donors. However, the proliferative capacity of patient-derived MSCs was significantly reduced. There was a significant reduction in in vitro chondrogenic and adipogenic activity in cultures of patient-derived cells compared with that in normal cultures. There was no significant difference in in vitro osteogenic activity. There was no decline in chondrogenic potential with age in cells obtained from individuals with no evidence of OA. CONCLUSION: These results raise the possibility that the increase in bone density and loss of cartilage that are characteristic of OA may result from changes in the differentiation profile of the progenitor cells that contribute to the homeostatic maintenance of these tissues. PMID- 11920407 TI - The PedsQL in pediatric rheumatology: reliability, validity, and responsiveness of the Pediatric Quality of Life Inventory Generic Core Scales and Rheumatology Module. AB - OBJECTIVE: The Pediatric Quality of Life Inventory (PedsQL) is a modular instrument designed to measure health-related quality of life (HRQOL) in children and adolescents ages 2-18 years. The 23-item PedsQL 4.0 Generic Core Scales are multidimensional child self-report and parent proxy-report scales developed as the generic core measure to be integrated with the PedsQL disease-specific modules. The 22-item PedsQL 3.0 Rheumatology Module was designed to measure pediatric rheumatology-specific HRQOL. This study was undertaken to demonstrate the reliability, validity, and responsiveness of the PedsQL 4.0 Generic Core Scales and the PedsQL 3.0 Rheumatology Module in pediatric rheumatology. METHODS: The 4 PedsQL 4.0 Generic Core Scales (physical, emotional, social, and school functioning) and the 5 PedsQL 3.0 Rheumatology Module scales (pain and hurt, daily activities, treatment, worry, and communication) were administered to 231 children and 244 parents (271 subjects accrued overall) recruited from a pediatric rheumatology clinic. RESULTS: Internal consistency reliability for the PedsQL Generic Core total scale score (alpha = 0.91 for child self report, alpha = 0.93 for parent proxy report), physical health summary score (alpha = 0.87 for child self report, alpha = 0.89 for parent proxy report), and psychosocial health summary score (alpha = 0.86 for child self report, alpha = 0.90 for parent proxy report) were acceptable for group comparisons. The Rheumatology Module scales also demonstrated acceptable reliability for group comparisons (alpha = 0.75-0.86 for child self report, alpha = 0.82-0.91 for parent proxy report). Validity was demonstrated using the known-groups method. The PedsQL distinguished between healthy children and children with rheumatic diseases as a group. The responsiveness of the PedsQL was demonstrated through patient change over time as a result of clinical intervention. CONCLUSION: The results demonstrate the reliability, validity, and responsiveness of the PedsQL 4.0 Generic Core Scales and the PedsQL 3.0 Rheumatology Module in pediatric rheumatology. PMID- 11920408 TI - Autoantibody to hLSm4 and the heptameric LSm complex in anti-Sm sera. AB - OBJECTIVE: To characterize the 15-kd human SmD-like autoantigen and its associated proteins previously shown to be recognized by IgM antibodies in patients with Epstein-Barr virus (EBV)-induced infectious mononucleosis. METHODS: The full-length complementary DNA for the 15-kd protein was expressed as recombinant protein and analyzed for reactivity using biochemical analysis and immunoprecipitation (IP). RESULTS: The 15-kd protein was determined to be the human like-Sm protein LSm4 (hLSm4). Rabbit antibody raised against the C-terminal polypeptide immunoprecipitated a 68-kd complex composed of LSm4 together with a group of smaller proteins ranging in size from 6.5 to 14 kd, consistent with the reported heptameric LSm complexes involved in U4/U6 duplex formation and messenger RNA (mRNA) decapping/degradation. About 80% of all anti-Sm sera from patients with systemic lupus erythematosus (SLE) recognized the hLSm4 in vitro translated product, while 6.7% (29 of 434) immunoprecipitated from cell extracts hLSm4 together with the other members of the hLSm complex. Four sera (0.92%) showed apparently exclusive reactivity to the hLSm complex in the absence of reactivity to Sm core proteins in the IP assay. CONCLUSION: These findings document that while IgM, but not IgG, autoantibodies to LSm4 were found in sera from patients with EBV infection, IgG autoantibodies to hLSm4 are detected in a large number of anti-Sm-positive sera from patients with SLE. Importantly, in a small number of anti-Sm sera the LSm complex can be recognized independently of the Sm core protein antigens. Our data introduce the concept that "Sm" autoantigens include Sm as well as LSm complexes involved in the maturation and degradation of mRNA. PMID- 11920409 TI - Selective accumulation of CCR4+ T lymphocytes into renal tissue of patients with lupus nephritis. AB - OBJECTIVE: Some chemokine receptors, such as CCR5 and CCR4, are differentially expressed on Th1 and Th2 cells. To determine whether differential expression of the chemokine receptors occurs in patients with lupus nephritis, we examined the expression of CCR4 and CCR5 on peripheral blood lymphocytes and mononuclear cells infiltrated into the renal tissue of patients with lupus nephritis. METHODS: The expression of CCR4 and CCR5 on CD4+,CD45RO+ cells was analyzed by flow cytometry and compared between patients with systemic lupus erythematosus (SLE) and healthy controls. Correlation between the absolute number of CCR4+ or CCR5+ cells and clinical parameters was also analyzed. Mononuclear infiltrates in the renal tissue of SLE patients were analyzed for the expression of CCR4, CCR5, and CD4 by immunohistochemical staining. RESULTS: The absolute number of CCR4+, but not CCR5+, T lymphocytes in the peripheral blood was significantly decreased in the patients with SLE compared with that in the healthy controls, and this positively correlated with the serum levels of C3 and CH50. Most of the CD4+ T lymphocytes that infiltrated into the renal tissue of the patients with lupus nephritis expressed CCR4, but not CCR5. CONCLUSION: These results suggest that CCR4+ T lymphocytes in peripheral blood, which represent Th2 cells, preferentially migrate into the renal tissue of patients with lupus nephritis. The maldistribution of CCR4+ T lymphocytes might be involved in the pathogenesis of lupus nephritis. PMID- 11920410 TI - Long-term risk of mortality and lymphoproliferative disease and predictive classification of primary Sjogren's syndrome. AB - OBJECTIVE: Primary Sjogren's syndrome (SS) may lead to lymphoproliferative disease (LPD) and death in certain patients. We sought to determine the incidence and predictors of adverse long-term outcomes to achieve a rational predictive classification of the syndrome. METHODS: Predictive modeling was performed in a cohort of 723 consecutive patients with primary SS (587 newly diagnosed [incident] cases and 136 prevalent cases). RESULTS: During 4,384 person-years of followup, we recorded 39 deaths (7 due to lymphoma) and 38 diagnoses of LPD. The standardized mortality ratio was 1.15 (95% confidence interval [95% CI] 0.86 1.73) compared with the general population of Greece. In incident cases, the probability of LPD was 2.6% at 5 years and 3.9% at 10 years. Mortality rates were significantly higher in patients with low C4 levels at the first study visit (hazard ratio [HR] 4.39, 95% CI 2.18-8.83). LPD was independently predicted by the presence of parotid enlargement (HR 5.21, 95% CI 1.76-15.4), palpable purpura (HR 4.16, 95% CI 1.65-10.5), and low C4 levels (HR 2.40, 95% CI 0.99-5.83) at the first study visit. All patients who eventually developed lymphoma resulting in death during the followup period had either low C4 levels or palpable purpura at the first study visit. Training-validation split-cohort modeling confirmed the predictive importance of low C4 levels and palpable purpura, both of which were present in 20.9% of patients at their first visit. CONCLUSIONS: In patients with primary SS, 1 in 5 deaths is attributable to lymphoma. The presence of palpable purpura and low C4 levels at the first visit adequately distinguishes high-risk patients (type I primary SS) from patients with an uncomplicated disease course (type II [low-risk] primary SS). PMID- 11920411 TI - A double-blind, randomized, placebo-controlled study of cevimeline in Sjogren's syndrome patients with xerostomia and keratoconjunctivitis sicca. AB - OBJECTIVE: To evaluate the safety and efficacy of 2 dosages of cevimeline for the treatment of xerostomia and keratoconjunctivitis sicca in patients with Sjogren's syndrome. METHODS: A 12-week double-blind, randomized, placebo-controlled study was performed. Patients were randomly assigned to receive either placebo, 15 mg of cevimeline 3 times daily, or 30 mg of cevimeline 3 times daily. Patients were evaluated at baseline and throughout the study for their global assessment of dryness (mouth, eyes, overall) as well as their subjective assessment of the specific symptoms of dry mouth and dry eyes. Total saliva and tear flow also were measured. RESULTS: Patients taking 30 mg of cevimeline 3 times daily had statistically significant improvements in their subjective global assessment of dry eyes (P = 0.0453), dry mouth (P = 0.0004), and increased salivary flow (P = 0.007). Patients receiving the 30-mg dosage also showed greater objective improvement (increased salivary and lacrimal flow rates, as measured by Schirmer's test) than did patients receiving placebo. Frequently reported adverse events included headache, increased sweating, abdominal pain, and nausea. CONCLUSION: Treatment with cevimeline at a dosage of 30 mg 3 times daily resulted in substantive improvement by increasing the rate of saliva and tear flow in patients with Sjogren's syndrome, as well as improving subjective symptoms of dry mouth, dry eyes, and overall dryness. The 15-mg dosage relieved some symptoms, and both dosages were well tolerated. PMID- 11920412 TI - Randomized double-blind comparison of chimeric monoclonal antibody to tumor necrosis factor alpha (infliximab) versus placebo in active spondylarthropathy. AB - OBJECTIVE: To confirm in a placebo-controlled trial the safety and efficacy profile of infliximab in short-term treatment of patients with active spondylarthropathy (SpA). METHODS: Forty patients with active SpA were randomly assigned to receive an intravenous loading dose (weeks 0, 2, and 6) of 5 mg/kg infliximab or placebo. Evaluations for efficacy and safety were performed at weeks 1, 2, 6, 8, and 12. The primary end points of this study were the improvements in patient and physician global assessments of disease activity on a 100-mm visual analog scale. RESULTS: Both primary end points improved significantly in the infliximab group compared with the baseline value, with no improvement in the placebo group. As early as week 2 and sustained up to week 12, there was a highly statistically significant difference between the values for these 2 end points in the infliximab versus the placebo group. In most of the other assessments of disease activity (laboratory measures, assessments of specific peripheral and/or axial disease), significant improvements were observed in the infliximab group compared with the baseline value and compared with placebo. Minor adverse events not causing discontinuation were equally observed in both treatment groups. There was one severe drug-related adverse event, in which a patient developed disseminated tuberculosis. CONCLUSION: Tumor necrosis factor alpha blockade with infliximab in patients with active SpA was well tolerated and resulted in significant clinical and laboratory improvements in this short-term, placebo-controlled study. However, the occurrence of tuberculosis in one patient necessitates strict inclusion criteria and long-term followup. PMID- 11920413 TI - A six-month randomized, controlled, double-blind, dose-response comparison of intravenous pamidronate (60 mg versus 10 mg) in the treatment of nonsteroidal antiinflammatory drug-refractory ankylosing spondylitis. AB - OBJECTIVE: To determine the safety and efficacy of intravenous (IV) pamidronate treatment in ankylosing spondylitis (AS) patients who have had a suboptimal response to nonsteroidal antiinflammatory drugs (NSAIDs). METHODS: Pamidronate at 60 mg was compared with pamidronate at 10 mg rather than placebo in view of the high incidence of transient arthralgias upon first IV exposure to the drug. The drug were given monthly for 6 months in a randomized double-blind, controlled trial. The inclusion criterion was active disease (Bath AS Disease Activity Index [BASDAI] of > or = 4 or morning stiffness of > or = 45 minutes) despite stable NSAID therapy. The primary outcome measure was the BASDAI, and secondary outcomes included the Bath AS Functional Index (BASFI), Bath AS Global Index (BASGI), Bath AS Metrology Index (BASMI), erythrocyte sedimentation rate (ESR), C-reactive protein (CRP) level, and percentage of patients achieving a reduction of > or = 25% in the BASDAI. Outcome assessments were done at -2, 0, 12, and 24 weeks, and analysis was by intent to treat. RESULTS: Eighty-four AS patients (67 men and 17 women; mean age 39.6 years and mean disease duration 15.1 years) were enrolled. Dosage groups were well matched at baseline for demographics, disease activity, and functional indices. At 6 months, the mean BASDAI had decreased by 2.22 (34.5%) in the 60-mg group and by 0.93 (15%) in the 10-mg group (P = 0.002). Significantly greater reductions in the 60-mg group were also noted for the BASFI (P < 0.001), BASGI (P = 0.01), and BASMI (P = 0.03). Significantly more patients achieved a reduction of > or = 25% in the BASDAI in the 60-mg group versus the 10 mg group (63.4% versus 30.2%; P = 0.004). Differences in ESR/CRP were not significant (NS). Withdrawals included 9 (20.9%) from the 10-mg group and 3 (7.3%) from the 60-mg group (P NS). Adverse events were confined to transient arthralgias/myalgias after the first IV infusion, occurring in 68.3% and 46.5% of patients in the 60-mg and 10-mg groups, respectively (P NS). CONCLUSION: Pamidronate has dose-dependent therapeutic properties in AS. PMID- 11920414 TI - Investigation of fluoroquinolone-induced myalgia using (31)P magnetic resonance spectroscopy and in vitro contracture tests. AB - OBJECTIVE: To investigate muscle function in patients with severe myalgia resulting from fluoroquinolone (FQ) treatment. We used histology, in vitro contracture tests (IVCTs), and (31)P magnetic resonance spectroscopy ((31)P MRS) to explore muscle contraction and metabolism. METHODS: We studied 3 patients with myalgia, hyperalgia tendinopathy, and arthralgia following FQ treatment and 3 normal subjects after taking FQs. Results were compared with those of a control group of 9 subjects free of any muscle disease and not taking FQs. Muscle biopsies were performed on the left biceps, and IVCTs were performed in accordance with the protocol recommended by the European Malignant Hyperthermia Group. (31)P MR spectra of forearm flexor muscles were recorded at 4.7T throughout a rest-exercise-recovery protocol. RESULTS: (31)P MRS showed a significant reduction of pH changes measured at the end of exercise and a faster rate of proton efflux measured during recovery in all patients. IVCTs diagnosed 1 patient as being susceptible to malignant hyperthermia. No specific histologic anomalies were observed in muscle biopsy samples, which showed normal mitochondria. CONCLUSION: The adverse effects recorded in the 3 patients are related to a preexisting muscular anomaly revealed by FQ treatment. PMID- 11920415 TI - Secular changes in the quality of published randomized clinical trials in rheumatology. AB - OBJECTIVE: To assess the quality of published randomized clinical trials (RCTs) in rheumatology and to determine whether there has been improvement in quality between 2 time periods, 1987-1988 and 1997-1998. METHODS: Using MEDLINE and a hand search of selected rheumatology journals, we identified RCTs of adult rheumatic diseases published in English in 1987-1988 or 1997-1998. We examined trial quality with an expanded version of the Jadad scale, which assesses the adequacy of reported random sequence generation, allocation concealment, blinding, and analysis. All trials were read by 1 reviewer, with prior standardization using a random sample read by 2 reviewers. We also evaluated "high"- versus "low"-impact journals based on citation index. RESULTS: Two hundred forty RCTs (1987-1988 119 RCTs, 1997-1998 121 RCTs) were assessed. Results showed improvement in the quality of the trials, but the rates of reported random sequence generation, allocation concealment, power, and intent-to treat analyses were persistently low. Low rates of reports of random sequence generation, allocation concealment, and intent-to-treat analyses were present even in the high-impact journals. CONCLUSION: There has been improvement in the quality of reporting of RCTs in rheumatology between 1987-1988 and 1997-1998. However, methodologic problems such as lack of allocation concealment, inadequate random sequence generation, lack of reporting of power, and lack of intent-to treat analyses remain common. Many of these problems are established sources of bias in RCTs and are easily rectifiable. PMID- 11920416 TI - Tumor necrosis factor alpha-mediated joint destruction is inhibited by targeting osteoclasts with osteoprotegerin. AB - OBJECTIVE: To study the effects of osteoclast-targeted therapies, such as osteoprotegerin (OPG) and pamidronate, on joint inflammation and bone destruction using a tumor necrosis factor alpha (TNF alpha)-transgenic mouse model. METHODS: Mice were placed into 5 groups that received either OPG, pamidronate, a combination of both agents, infliximab as a positive control, or phosphate buffered saline as a negative control. Treatment was initiated at the onset of arthritis, continued over 6 weeks, and thereafter, the clinical, radiologic, and histologic outcomes were assessed. RESULTS: A significant improvement in clinical symptoms, as assessed by the reduction of paw swelling, was only found in the infliximab group, whereas all other treatment groups failed to show significant improvement. However, when assessing structural damage with radiographic analysis, a significant retardation of joint damage was evident in animals treated with OPG (55% reduction of erosions), pamidronate (50% reduction of erosions) the combination therapy of OPG and pamidronate (64% reduction of erosions), and with infliximab (66% reduction of erosions). Confirming these data, quantitative histologic analysis revealed a significant reduction in the size of bone erosions in all treatment groups (OPG 56%, pamidronate 53%, OPG and pamidronate 81%, and infliximab 46%) compared with the control group. Furthermore, a significant reduction of osteoclast numbers was seen in animals treated with OPG alone or in combination with pamidronate as well as in animals treated with infliximab. CONCLUSION: These data suggest that OPG alone or in combination with bisphosphonates is an effective therapeutic tool for the prevention of TNF alpha-mediated destruction of bone by reducing the number of bone-resorbing cells in the inflammatory tissue. PMID- 11920417 TI - Angiostatin gene transfer as an effective treatment strategy in murine collagen induced arthritis. AB - OBJECTIVE: To determine the efficacy of local therapy with human angiostatin gene in murine collagen-induced arthritis (CIA). METHODS: DBA/1 mice were immunized with bovine type II collagen. Before the onset of arthritis, NIH3T3 fibroblasts, transduced with angiostatin-expressing retroviral vectors or control vectors, were transplanted into the knee cavity. The incidence of arthritis in the knee joints was evaluated histologically based on pannus formation and cartilage destruction. Paws were evaluated macroscopically for redness, swelling, and deformities and immunologically for levels of interleukin-1 beta. Angiogenesis in paws and knee joints was studied by immunohistochemistry using anti-CD31 antibody and measurement of von Willebrand factor levels. RESULTS: Pannus formation and cartilage erosion were dramatically reduced in knees transplanted with angiostatin-expressing cells. In addition, the onset of CIA in the ipsilateral paws below the knees injected with the angiostatin gene was significantly prevented. Furthermore, angiostatin gene transfer inhibited arthritis-associated angiogenesis. CONCLUSION: Local production of angiostatin in the knee was able to prevent the onset of CIA not only in the knee injected with genetically engineered cells, but also in the uninjected ipsilateral paw. This suggests that transfer of the angiostatin gene, and potentially also its protein, may provide a new, effective approach to the treatment of rheumatoid arthritis. PMID- 11920418 TI - Reduction of joint inflammation and bone erosion in rat adjuvant arthritis by treatment with interleukin-17 receptor IgG1 Fc fusion protein. AB - OBJECTIVE: To investigate the role of interleukin-17 (IL-17) in inflammatory arthritis by blockade with an IL-17 receptor/human IgG1 Fc fusion protein (muIL 17R:Fc) in adjuvant-induced arthritis (AIA) in the rat. METHODS: AIA was induced in 39 DA rats with the use of Freund's complete adjuvant. Rats received either 7.3 or 20 mg/kg of muIL-17R:Fc or phosphate buffered saline intraperitoneally every other day from the time of arthritis induction for approximately 17 days. Paw volume, arthritis severity, and weight were assessed every 3-4 days. Rats were killed between days 21 and 23 post-induction. Ankles were removed for quantitative radiology and histology and for immunohistochemistry for T cells. RESULTS: Treatment with muIL-17R:Fc attenuated paw volume in a dose-dependent manner. Both the 7.3 and 20 mg/kg doses of muIL-17R:Fc significantly reduced radiographic scores in the treated rats compared with the controls. The 20 mg/kg dose of muIL-17R:Fc significantly reduced histology scores compared with the controls. T cell numbers were unchanged in the muIL-17R:Fc-treated rats as a function of dose. CONCLUSION: In vivo blockade of IL-17 by muIL-17R:Fc treatment attenuated AIA and reduced joint damage, suggesting that IL-17 plays an important role in the inflammation and joint destruction of AIA. IL-17 may be a potential therapeutic target for inflammatory diseases in humans, such as rheumatoid arthritis. PMID- 11920419 TI - The beneficial effect of interleukin-12 on arthritis induced by group B streptococci is mediated by interferon-gamma and interleukin-10 production. AB - OBJECTIVE: To assess the effect of interleukin-12 (IL-12) administration on the evolution of systemic infection and septic arthritis induced by group B streptococci (GBS) in mice. METHODS: CD1 mice were inoculated intravenously with arthritogenic strain 1/82 of type IV GBS. Exogenous murine IL-12 was administered intraperitoneally 18 hours or 5 days after infection with 1 x 10(7) GBS, at doses ranging from 0.5 to 2.5 microg per mouse. Mice were monitored daily for survival and for signs of arthritis. In a subsequent set of experiments, mice were killed at selected times for examination of bacterial clearance, histopathologic changes in the joints, and cytokine production. RESULTS: IL-12 administration before the onset of clinical signs had a beneficial effect on GBS-induced arthritis and was clearly dose-dependent. The 2.5-microg dose per mouse totally prevented death from GBS-induced arthritis. The decrease in pathology was associated with a reduction of the bacterial burden and a change in the cytokine profile. In particular, systemic and joint levels of interferon-gamma (IFN gamma) and IL-10 significantly increased in mice treated with IL-12, whereas a decrease in IL-6 and IL-1 beta production was observed. The beneficial effects of IL-12, in terms of the incidence and severity of articular lesions, were reversed by coadministration of anti-IFN gamma or anti-IL-10-neutralizing antibodies. CONCLUSION: The findings of this study demonstrate that IL-12 is important in controlling the cytokine production that leads to the evolution of GBS-induced experimental arthritis. The amelioration of articular lesions is mostly attributable to IL-12-induced IFN gamma, but with a relevant participation of IL 12-induced IL-10. PMID- 11920420 TI - Joint damage and inflammation in c-Jun N-terminal kinase 2 knockout mice with passive murine collagen-induced arthritis. AB - OBJECTIVE: Previous studies have demonstrated that inhibition of c-Jun N-terminal kinase (JNK) decreases joint destruction in the rat adjuvant arthritis model. The present study was undertaken to investigate whether selective loss of JNK-2 function decreases joint destruction in JNK-2 knockout mice, in order to determine the role of this isoform in inflammatory arthritis. METHODS: Passive collagen-induced arthritis (CIA) was induced in Jnk2(-/-) and wild-type mice by administering anti-type II collagen antibodies. Arthritis was assessed daily using a semiquantitative clinical scoring system. Fibroblast-like synoviocytes (FLS) were prepared from Jnk2(-/-) and wild-type mice, and JNK protein expression was determined by Western blot analysis. Matrix metalloproteinase 13 (MMP-13) expression was determined by Northern blot analysis, and activator protein 1 (AP 1) binding activity by electromobility shift assay (EMSA). RESULTS: The JNK protein level in Jnk2(-/-) mice with CIA was 22% of that in wild-type mice with CIA (P < 0.001), and mainly the 46-kd isoform was expressed in the former group. Surprisingly, clinical arthritis was slightly more severe in the Jnk2(-/-) mice. Histologic scores for synovial inflammation were not significantly different. However, Safranin O-stained sections from the Jnk2(-/-) mice exhibited significantly less joint damage. Although joint destruction was decreased in Jnk2(-/-) mice with CIA, EMSA and Northern blot analysis of total joint extracts revealed similar levels of AP-1 binding and MMP-13 expression in Jnk2(-/-) and wild-type mice. The lack of correlation with AP-1 activity and MMP expression was probably because non-FLS cells in the joint may express more JNK-1 than do FLS. CONCLUSION: JNK-2 is a determinant of matrix degradation, but it has little effect on inflammation in arthritis. Complete inhibition of MMP expression and joint destruction will likely require combined JNK-1 and JNK-2 inhibition. PMID- 11920421 TI - Stromal cell-derived factor 1 (CXCL12) induces monocyte migration into human synovium transplanted onto SCID Mice. AB - OBJECTIVE: The mechanisms by which monocyte/macrophage cells migrate to the joint involve a series of integrated adhesion and signaling events in which chemokines and their receptors are strongly implicated. This study was undertaken to investigate the hypothesis that stromal cell-derived factor 1 (SDF-1), a CXC chemokine (CXCL12), plays a critical role in monocyte/macrophage localization to synovium. METHODS: SDF-1 and CXC receptor 4 (CXCR4) expression in rheumatoid arthritis (RA) and osteoarthritis synovium and graft SDF-1, tumor necrosis factor alpha (TNF alpha), and human and murine vascular markers were examined by immunohistochemistry and double-immunofluorescence. The functional capacity of SDF-1 to modulate monocyte migration into joints was investigated by examining the localization of pro-myelomonocytic U937 cells into synovial tissue transplanted into SCID mice. SDF-1, TNF alpha, or saline was injected into graft sites and response determined by the number of fluorescently labeled U937 cells (injected intravenously) detected in grafts by ultraviolet microscopy. RESULTS: SDF-1 and CXCR4 were highly expressed in CD68+ cells in the RA synovium. SDF-1 induced U937 cell migration in vitro and in vivo in a dose-dependent manner and, in vivo, SDF-1 was more effective than TNF alpha. In contrast to TNF alpha, SDF-1 did not induce intracellular adhesion molecule 1 in transplant microvasculature. Furthermore, intragraft injection of SDF-1 did not up-regulate TNF alpha, or vice versa. CONCLUSION: This study demonstrates, for the first time, that SDF-1 is functional in vivo when injected into synovial grafts. In addition, SDF-1 is more potent than TNF alpha, and its mechanisms of action appear to be autonomous. Therefore, SDF-1 may be an important TNF-independent molecule involved in the migration to and retention of inflammatory effector cells in the joint. PMID- 11920422 TI - High-dose cyclophosphamide with stem cell rescue for severe rheumatoid arthritis: short-term efficacy correlates with reduction of macroscopic and histologic synovitis. PMID- 11920423 TI - Refractory inflammatory heel pain in spondylarthropathy: a significant response to infliximab documented by ultrasound. PMID- 11920425 TI - Development of rheumatoid nodules during anti-tumor necrosis factor alpha therapy with etanercept. PMID- 11920426 TI - Topotecan and the development of scleroderma or a scleroderma-like illness. PMID- 11920427 TI - Methotrexate for the treatment of early diffuse scleroderma: comment on the article by Pope et al. PMID- 11920428 TI - Confirmation of an association between fibromyalgia and serotonin transporter promoter region (5- HTTLPR) polymorphism, and relationship to anxiety-related personality traits. PMID- 11920429 TI - Effect of intraarticular hyaluronate injections in chondrocalcinosis: comment on the article by Martens. PMID- 11920431 TI - Inflammatory arthritis and the diagnosis and management of iron deficiency: comment on the article by Bultink et al. PMID- 11920433 TI - Absence of anti-cyclic citrullinated peptide antibodies in antineutrophil cytoplasmic antibody-associated vasculitis. PMID- 11920434 TI - Catastrophic antiphospholipid syndrome associated with typhoid fever: comment on the article by Hayem et al. PMID- 11920436 TI - Tetrazole analogues of cyclolinopeptide A: synthesis, conformation, and biology. AB - Linear and cyclic analogues of cyclolinopeptide A (CLA) with two dipeptide segments (Val(5)-Pro(6) and Pro(6)-Pro(7)) replaced by their tetrazole derivatives were synthesized by the SPPS technique and cyclized using TBTU (O (benzotriazol-1-yl)-1,1,3,3-tetramethyluronium tetrafluoroborate) reagent. The conformational properties of the c(Leu(1)-Ile(2)-Ile(3)-Leu(4)-Val(5)-Pro(6) psi[CN(4)]-Ala(7)-Phe(8)-Phe(9)) were investigated by NMR and computational techniques. The overall solution structure of this cyclic peptide resembles that observed for the CLA in the solid state. These studies of cyclic tetrazole CLA analogue confirm that the 1,5-disubstituted tetrazole ring functions as an effective, well-tolerated cis-amide bond mimic in solution. The peptides were examined for their immunosuppressive activity in the humoral response test. For cyclic analogues the immunosuppressive activity, at low doses, is equal in magnitude to the activity presented by cyclosporin A and native CLA. The conformational and biological data seem indicate that the Pro-Pro-Phe-Phe moiety and the preservation of the CLA backbone conformation are important for immunosuppressive activity. PMID- 11920437 TI - Model peptide studies of sequence regions in the elastomeric biomineralization protein, Lustrin A. I. The C-domain consensus-PG-, -NVNCT-motif. AB - The lustrin superfamily represents a unique group of biomineralization proteins localized between layered aragonite mineral plates (i.e., nacre layer) in mollusk shell. Recent atomic force microscopy (AFM) pulling studies have demonstrated that the lustrin-containing organic nacre layer in the abalone, Haliotis rufescens, exhibits a typical sawtooth force-extension curve with hysteretic recovery. This force extension behavior is reminiscent of reversible unfolding and refolding in elastomeric proteins such as titin and tenascin. Since secondary structure plays an important role in force-induced protein unfolding and refolding, the question is, What secondary structure(s) exist within the major domains of Lustrin A? Using a model peptide (FPGKNVNCTSGE) representing the 12 residue consensus sequence found near the N-termini of the first eight cysteine rich domains (C-domains) within the Lustrin A protein, we employed CD, NMR spectroscopy, and simulated annealing/minimization to determine the secondary structure preferences for this sequence. At pH 7.4, we find that the 12-mer sequence adopts a loop conformation, consisting of a "bend" or "turn" involving residues G3-K4 and N7-C8-T9, with extended conformations arising at F1-G3; K4-V6; T9-S10-G11 in the sequence. Minor pH-dependent conformational effects were noted for this peptide; however, there is no evidence for a salt-bridge interaction between the K4 and E12 side chains. The presence of a loop conformation within the highly conserved -PG-, -NVNCT- sequence of C1-C8 domains may have important structural and mechanistic implications for the Lustrin A protein with regard to elastic behavior. PMID- 11920438 TI - Ordering in aqueous polysaccharide solutions. II. Optical rotation and heat capacity of aqueous solutions of a triple-helical polysaccharide schizophyllan. AB - Deuterium oxide solutions of schizophyllan, a triple-helical polysaccharide, undergoing an order-disorder transition centered at 17 degrees C, were studied by optical rotation (OR) and heat capacity (C(p)) to elucidate the molecular mechanism of the transition and water structure in the solution and frozen states. The ordered structure at low temperature consisted of the side chains and water in the vicinity forming an ordered hydrogen-bonded network surrounding the helix core and was disordered at higher temperature. In the solution state appeared clearly defined transition curves in both the OR and C(p) data. The results for three samples of different molecular weights were analyzed theoretically, treating this transition as a typical linear cooperative transition from the ordered to disordered states and explained quantitatively if the molecular weight polydispersity of the sample was considered. The excess heat capacity C(EX)(p) defined as the C(p) minus the contributions from schizophyllan and D(2)O was estimated. In the frozen state it increased with raising temperature above 150 K until the mixture melted. This was compared with the dielectric increment observed in this temperature range and ascribed to unfreezable water. From the heat capacity and dielectric data, unfreezable water is mobile but more ordered than free water. In the solution state, the excess heat capacity originates from the interactions of D(2)O molecules as bound water and structured water, and so forth. Thus the schizophyllan triple helix molds water into various structures of differing orders in solution and in the solid state. PMID- 11920439 TI - Photomodulation of conformational states. III. Water-soluble bis-cysteinyl peptides with (4-aminomethyl) phenylazobenzoic acid as backbone constituent. AB - In previous studies we have shown that light-induced cis/trans isomerization of the azobenzene moiety in cyclo-[Ala-Cys-Ala-Thr-Cys-Asp-Gly-Phe-AMPB] [AMPB: (4 aminomethyl)phenylazobenzoic acid] leads both in the monocyclic and in the oxidized bicyclic form to markedly differentiated conformational states in DMSO, a fact that lends itself for photomodulation of the redox potential of such bis cysteinyl-peptides. For this purpose water-soluble systems are required, and this was achieved by replacing three residues outside the Cys-Ala-Thr-Cys active-site motif of thioredoxin reductase with lysines. The resulting cyclo-[Lys-Cys-Ala-Thr Cys-Asp-Lys-Lys-AMPB] fully retains its photoresponsive properties in water as well assessed by uv and CD measurements. Paralleling results of the previously investigated azobenzene-containing cyclic peptides, the trans --> cis isomerization of the water-soluble monocyclic and oxidized bicyclic peptide is accompanied by a marked transition from a well-defined conformation to an ensemble of possible conformations. However, the conformational preferences are very dissimilar from those of the DMSO-soluble peptides. In fact, hydrogen bonds as well as secondary structure elements were found that change in the mono- and bicyclic peptide upon irradiation. The photo switch between different turn types and hydrogen bonding networks offers the structural rational for the significantly differentiated redox potentials, but also the possibility of monitoring by femtosecond uv-vis and ir spectroscopy fast and ultra fast backbone rearrangement processes following the electronic trans --> cis isomerization. PMID- 11920440 TI - Albumin recovery with centrifugal adsorption technology (CAT). AB - Centrifugal adsorption technology (CAT) is a new compact, countercurrent technology for efficient adsorption from large liquid streams by using adsorbent particles in the micrometer range. CAT seems particularly suited for the recovery of macromolecules at low concentrations, because the small particle dimensions lead to fast mass transfer rates. In this work, the potential of CAT for protein recovery is studied by model and experiment. A predictive model for the separation performance of CAT is presented, incorporating mass transfer resistance and axial dispersion transport in the liquid and the adsorbent phases. The model calculations were compared to experimental data for the adsorption of bovine serum albumin (BSA) on a standard commercial anion-exchange resin with particle diameter d(p) = 50 microm in a pilot-scale CAT apparatus. The model calculations accurately predicted the separation efficiency of CAT. The experimental set-up is shown to be mass transfer limited for the conducted experiments, which agrees with the model predictions. The model was also used to estimate the dimensions and performance of a CAT apparatus for the large-scale recovery of human serum albumin (HSA) from fermentation broth at the scale of 40 tons per year. The resulting equipment dimensions proved to be very small indeed, making CAT a potentially very attractive technology. PMID- 11920441 TI - Direct transesterification of gases by "dry" immobilized lipase. AB - Several different reactor configurations, including single pass, continuous recycle, and batch reactor modes, were used to investigate the effects of temperature and water activity, or relative humidity, on lipase-catalyzed, gas phase transesterifications. Temperature and relative humidity were controlled both inside reactors and throughout the course of the reaction to account for and optimize their effects. Results indicated that, at low relative humidity, reaction rates increased with temperature up to 60 degrees C. However, when relative humidity was increased, a similar increase in temperature resulted in the loss of nearly all enzyme activity. These results are consistent with the idea that enzymes without free water are more thermally stable. Furthermore, at constant ambient temperatures, production increased dramatically with an increase in relative humidity, confirming the idea that an increase in water activity increases catalytic activity. A mass balance performed on reactors at higher relative humidity revealed that hydrolysis (rather than transesterification) of the ester substrate could significantly decrease product yields. PMID- 11920442 TI - A microfabricated array bioreactor for perfused 3D liver culture. AB - We describe the design, fabrication, and performance of a bioreactor that enables both morphogenesis of 3D tissue structures under continuous perfusion and repeated in situ observation by light microscopy. Three-dimensional scaffolds were created by deep reactive ion etching of silicon wafers to create an array of channels (through-holes) with cell-adhesive walls. Scaffolds were combined with a cell-retaining filter and support in a reactor housing designed to deliver a continuous perfusate across the top of the array and through the 3D tissue mass in each channel. Reactor dimensions were constructed so that perfusate flow rates meet estimated values of cellular oxygen demands while providing fluid shear stress at or below a physiological range (<2 dyne cm(2)), as determined by comparison of numerical models of reactor fluid flow patterns to literature values of physiological shear stresses. We studied the behavior of primary rat hepatocytes seeded into the reactors and cultured for up to 2 weeks, and found that cells seeded into the channels rearranged extensively to form tissue like structures and remained viable throughout the culture period. We further observed that preaggregation of the cells into spheroidal structures prior to seeding improved the morphogenesis of tissue structure and maintenance of viability. We also demonstrate repeated in situ imaging of tissue structure and function using two-photon microscopy. PMID- 11920444 TI - Optimizing the heterologous production of epothilone D in Myxococcus xanthus. AB - The heterologous production of epothilone D in Myxococcus xanthus was improved by 140-fold from an initial titer of 0.16 mg/L with the incorporation of an adsorber resin, the identification of a suitable carbon source, and the implementation of a fed-batch process. To reduce the degradation of epothilone D in the basal medium, XAD-16 (20 g/L) was added to stabilize the secreted product. This greatly facilitated its recovery and enhanced the yield by three-fold. The potential of using oils as a carbon source for cell growth and product formation was also evaluated. From a screen of various oils, methyl oleate was shown to have the greatest impact. At the optimal concentration of 7 mL/L in a batch process, the maximum cell density was increased from 0.4 g dry cell weight (DCW)/L to 2 g DCW/L. Product yield, however, depended on the presence of trace elements in the production medium. With an exogenous supplement of trace metals to the basal medium, the peak epothilone D titer was enhanced eight-fold. This finding demonstrates the significant role of metal ions in cell metabolism and in epothilone biosynthesis. To further increase the product yield, a continuous fed batch process was used to promote a higher cell density and to maintain an extended production period. The optimized fed-batch cultures consistently yielded a cell density of 7 g DCW/L and an average production titer of 23 mg/L. PMID- 11920443 TI - Pilot-scale process sensitivity studies for the scaleup of a fungal fermentation for the production of pneumocandins. AB - The filamentous fungus Glarea lozoyensis produces a novel, pharmaceutically important pneumocandin (B(0)) that is used to synthesize a lipopeptide which demonstrates cidal activity against clinically relevant pathogens. A range of unwanted pneumocandin analogs are also produced by the organism. To maintain the unwanted impurities to acceptable levels upon scaleup, a good understanding of the impact of chemical and physical environment on the cell physiology is required, which benefits downstream processing. Pilot-scale studies were performed to determine the impact of dissolved oxygen, temperature, pH, and carbon dioxide on the process. Experiments included multiple fermenters (up to seven) at 0.07 and 0.8 m(3) scale using single source medium sterilization and inoculum. Gas blending was used to separate effects of dissolved oxygen from agitation. The process was significantly influenced by dissolved oxygen level. The critical dissolved oxygen tension (C(crit)) for growth was below 2% air saturation. The C(crit) for production of pneumocandin B(0) was 20% air saturation, with a significant reduction of the specific production rate below this value. In contrast, low dissolved oxygen levels produced a substantial increase of pneumocandins B(1), B(5), and E(0), while high dissolved oxygen levels produced a disproportionate increase of D(5). This sensivity to dissolved oxygen was independent of agitation within a power range of 2-15 kW/m(3). Broth viscosity was impacted below 10% dissolved oxygen, suggesting an effect on morphology. The process was shown to be sensitive to temperature but relatively insensitive to pH and carbon dioxide (in the exhaust gas) within the ranges studied. This scaledown analysis explained phenomena seen at pilot scale and helped define operating boundary conditions for successful scale up to 19 m(3). PMID- 11920445 TI - Kinetic constants determination for an alkaline protease from Bacillus mojavensis using response surface methodology. AB - The kinetic constants for an alkaline protease from Bacillus mojavensis were determined using a central composite circumscribed design (CCCD) where concentration of substrate (casein) and the assay temperature were varied around their center point. The K(m),V(max), K(cat), activation energy (E(a)) and temperature coefficient (q(10)) were determined and the values of these kinetic constants obtained were found comparable to that obtained with conventional methods. The Michaelis-Menten constant (K(m)) for casein decreased with corresponding increase in V(max), as reaction temperature was raised from 45-60 degrees C. The protease exhibited K(m) of 0.0357 mg/ml, 0.0270 mg/ml, 0.0259 mg/ml, and 0.0250 mg/ml at 45, 50, 55, and 60 degrees C, respectively, whereas V(max) values at these temperatures were 74.07, 99.01, 116.28, and 120.48 microg/ml/min, respectively, as determined by response surface methodology. The Arrhenius plot suggested that the enzyme undergoes thermal activation above 45 degrees C until 60-65 degrees C followed by thermal inactivation. Likewise, the energy of activation (E(a)) was more between 45-55 degrees C (9747 cal/mol) compared to E(a) between 50-60 degrees C (4162 cal/mol). PMID- 11920446 TI - Stoichiometric model for evaluating the metabolic capabilities of the facultative methylotroph Methylobacterium extorquens AM1, with application to reconstruction of C(3) and C(4) metabolism. AB - A stoichiometric model of central metabolism was developed based on new information regarding metabolism in this bacterium to evaluate the steady-state growth capabilities of the serine cycle facultative methylotroph Methylobacterium extorquens AM1 during growth on methanol, succinate, and pyruvate. The model incorporates 20 reversible and 47 irreversible reactions, 65 intracellular metabolites, and experimentally-determined biomass composition. The flux space for this underdetermined system of equations was defined by finding the elementary modes, and constraints based on experimental observations were applied to determine which of these elementary modes give a reasonable description of the flux distribution for each growth substrate. The predicted biomass yield, on a carbon atom basis, is 49.8%, which agrees well with the range of published experimental yield measurements (37-50%). The model predicts the cell to be limited by reduced pyridine nucleotide availability during methylotrophic growth, but energy-limited when growing on multicarbon substrates. Mutation and phenotypic analysis was used to explore a previously unknown region of the metabolic map and to confirm the stoichiometry of the pathways in this region used in the metabolic model. Based on genome sequence data and simulation results, three enzymes involved in C(3)-C(4) interconversion pathways were predicted to be mutually redundant: malic enzyme, phosphoenolpyruvate carboxykinase, and phosphoenolpyruvate synthase. Insertion mutations in the genes predicted to encode these enzymes were made and these mutants were capable of growing on all substrates tested, confirming the redundancy of these pathways. Likewise, pathway analysis suggests that the TCA cycle enzymes citrate synthase and succinate dehydrogenase are essential for all growth substrates. In keeping with these predictions, null mutants could not be obtained in these genes. Finally, a similar model was developed for the ribulose monophosphate pathway obligate methylotroph Methylobacillus flagellatum KT to compare the efficiency of carbon utilization in the two types of methylotrophic carbon utilization pathways. The predicted yield for this organism on methanol is 65.9%. PMID- 11920447 TI - Esterification by immobilized lipase in solvent-free media: kinetic and thermodynamic arguments. AB - The aim of this study is to characterize, in solvent-free systems (SFS), the kinetic and thermodynamic performance of batch lipase-catalyzed esterification. SFS are compared to a conventional organic solvent, n-hexane. The esterification of oleic acid with ethanol was chosen as a model reaction. The TABEK (thermodynamic activity-based enzyme kinetics) approach was used to rationally analyze kinetics. Influence of the reaction medium on final conversions was also studied. Several factors, such as initial molar ratio of substrates, reactant availability, initial water content, and quantity of immobilized enzyme, were examined. Special attention was also turned to enzyme stability and reuse after reaction, this last item being a prerequisite in the development of industrial processes. SFS proved to be almost as efficient as n-hexane from a kinetic and thermodynamic point of view and offered a better volumetric production. PMID- 11920448 TI - A kinetic model describing cell growth and production of highly active, recombinant ice nucleation protein in Escherichia coli. AB - A structured kinetic model, which describes the production of the recombinant ice nucleation protein in different conditions, was applied. The model parameters were estimated based on the variation of the specific growth rate and the intracellular product concentration during cultivation. The equations employed relate the cellular plasmid content or plasmid copy number with the cloned-gene expression; these correlations were successfully tested on the experimental data. The optimal nutrient conditions for the growth of Escherichia coli expressing the inaZ gene of Pseudomonas syringae were determined for the production of active ice nucleation protein. The kinetics of the cultures expressing the inaZ gene were studied in a bioreactor at different growth temperatures and nutrient conditions. PMID- 11920449 TI - A simple method to release polyphosphate from activated sludge for phosphorus reuse and recycling. AB - In enhanced biological phosphorus removal processes, activated sludge microorganisms accumulate large quantities of polyphosphate (polyP). It was discovered that nearly all of the polyP could be released from activated sludge simply by heating it at 70 degrees C for about 1 h. The chain length of released polyP ranged from 100 to 200 phosphate (P(i)) residues. The addition of CaCl(2) precipitated approximately 75% of the total phosphorus without pH adjustment. The formed precipitate contained more P and less Ca than typical natural phosphorite deposits. Hence, in combination with enhanced biological phosphorus removal, the present method has potential for the development of a simple process for recovering phosphorus in a reusable form from wastewater. PMID- 11920450 TI - Engineering of baker's yeasts, E. coli and Bacillus hosts for the production of Bacillus subtilis Lipase A. AB - Lipases are versatile biocatalists showing multiple applications in a wide range of biotechnological processes. The gene lipA coding for Lipase A from Bacillus subtilis was isolated by PCR amplification, cloned and expressed in Escherichia coli, Saccharomyces cerevisiae and Bacillus subtilis strains, using pBR322, YEplac112 and pUB110-derived vectors, respectively. Lipase activity analysis of the recombinant strains showed that the gene can be properly expressed in all hosts assayed, this being the first time a lipase from bacterial origin can be expressed in baker's S. cerevisiae strains. An important increase of lipase production was obtained in heterologous hosts with respect to that of parental strains, indicating that the described systems can represent a useful tool to enhance productivity of the enzyme for biotechnological applications, including the use of the lipase in bread making, or as a technological additive. PMID- 11920451 TI - Coordination of cell growth in cocultures by a genetic proliferation control system. AB - Advanced cell culture systems such as organotypic cultures or stem cell expansion often involve cocultivation of two or more different cell types. A typical situation is the use of a feeder cell line that supports survival, growth or controls the differentiation status of primary cells. Unequal proliferation capacities result in unbalanced cell growth, such as the undesired expansion of immortalized cells at the expense of the primary cells. We utilized a genetic regulatory system to coordinate cell growth in cocultures. Proliferation control of a feeder cell line by a recombinant transcription factor was employed to prevent feeder cells from overgrowing cocultured stem cells, thereby avoiding premature media exhaustion. Relevant characteristic cell-surface proteins present on the parental feeder cell line were preserved. No special equipment was required and cell-type specific growth could be adjusted even during coculture. When compared to a conventional gamma-irradiation procedure, genetic proliferation control was superior with respect to ease of use, robustness, cell adherence, morphology and viability. The results demonstrate the applicability of genetic proliferation control to coordinate growth of different cell types in cocultures. PMID- 11920453 TI - Clinical pretreatment risk factors and Ga-67 scintigraphy early during treatment for prediction of outcome of patients with aggressive non-Hodgkin lymphoma. AB - BACKGROUND: Clinical pretreatment risk factors indicate the severity of disease in patients with aggressive non-Hodgkin lymphoma (NHL). Ga-67 scintigraphy during treatment is an early indicator of treatment-related features of lymphoma cells. The ability of risk factors and Ga-67 to predict disease outcome was compared in 139 patients with aggressive NHL. METHODS: Pretreatment clinical risk factors and Ga-67 scintigraphy performed after one cycle and at mid-treatment were evaluated for their correlation with response rate and as predictors of 5-year failure-free survival (FFS). Univariate analysis was performed to determine the ability of pretreatment risk factors and Ga-67 early during treatment to predict FFS. Subsequently, multivariate analysis was performed on the variables with significant univariate results using the Cox proportional hazards method. The predictive value of risk factors and Ga-67 scintigraphy was calculated to determine their suitability in selecting patients with poor outcome. RESULTS: Response rate correlated with stage of disease (P < 0.01) and the international prognostic index (IPI) score (P < 0.05). Five-year FFS was predicted by stage of disease (P < 0.004), performance status (P < 0.02), and the IPI score (P < 0.01). Response rate correlated with results of Ga-67 scintigraphy after one cycle of chemotherapy (P < 0.001) and at mid-treatment (P < 0.001). Five-year FFS was predicted by Ga-67 after one cycle of chemotherapy (P < 0.0004) and at mid treatment (P < 0.0001). Positive Ga-67 after the first cycle of treatment predicted 64% of patients who had failure of treatment. A positive study at mid treatment predicted 77% of patients who had treatment failure. Cox analysis showed Ga-67 after one course (P < 0.0012) and at mid-treatment (P < 0.0002) as being the most significant variables in predicting FFS. CONCLUSIONS: Ga-67 scintigraphy demonstrates early the effect of treatment in patients with aggressive NHL. It is a better predictor than pretreatment risk factors of both response rate and FFS. Positive Ga-67 early during treatment may be used as an independent test in selecting patients who will not respond favorably to current protocol treatment for early therapeutic modifications. PMID- 11920454 TI - Comparison of fluorine-18 fluorodeoxyglucose positron emission tomography and Ga 67 scintigraphy in evaluation of lymphoma. AB - BACKGROUND: The accuracy of fluorodeoxyglucose positron emission tomography (FDG PET; dual-head camera with attenuation correction) and Ga-67 scintigraphy was compared to identify disease sites in patients with Hodgkin disease (HD) and intermediate and high-grade non-Hodgkin lymphoma (NHL) at initial diagnosis or clinical recurrence. METHODS: Fifty-one contemporaneous FDG-PET and Ga-67 scintigraphies were performed on patients with NHL (35 intermediate grade, 3 high grade) or HD (13 patients). Sites of disease were correlated on a site-by-site basis on FDG-PET and Ga-67 images. Tumor-to-background (T/B) ratios were obtained for both techniques. Discordant FDG-PET and Ga-67 findings were correlated with computed tomography findings or clinical evaluation including repeat FDG-PET scans obtained after therapy. RESULTS: Fluorodeoxyglucose positron emission tomography was positive at all 158 sites in 51 patients compared with 113 sites in 41 positive studies with Ga-67 scintigraphy (single positron emission computed tomography [SPECT] and/or planar images). In 44 patients who had complete Ga-67 SPECT data on all tumor sites, FDG-PET was positive at 126 sites and Ga-67 SPECT was positive at 81 sites. Ga-67 SPECT failed to demonstrate disease at 45 sites (35.7%). In 10 of 44 patients, Ga-67 SPECT completely failed to detect any disease at 22 of 45 sites (17.5%) and partially identified disease sites at 23 of 45 sites (18.2%) in 11 patients regardless of the tumor site and histology. In these patients, the lesions measured between 0.6 and 14.0 cm by CT. Fluorodeoxyglucose positron emission tomography revealed higher stage disease in 13 patients compared with Ga-67 imaging. Tumor-to-background ratios were statistically different between the two techniques with higher ratios obtained with FDG-PET (P < 0.0001). CONCLUSIONS: In imaging aggressive lymphoma and HD before therapy, FDG-PET has significantly higher site and patient sensitivity than Ga-67 scintigraphy (100% vs. 71.5% and 100% vs. 80.3%, respectively). The change in disease stage by FDG-PET may result in a change in therapy strategy. PMID- 11920455 TI - Results of the Lynn Sage Second-Opinion Program for local therapy in patients with breast carcinoma. Changes in management and determinants of where care is delivered. AB - BACKGROUND: The treatment options of breast conservation therapy (BCT) and immediate reconstruction for patients with carcinoma of the breast have not been adopted widely. The objectives of this study were to determine how often a second opinion on the local therapy of breast carcinoma changed patient management and to identify factors predictive of remaining at the second-opinion site for therapy. METHODS: Two hundred thirty-one patients with intraductal carcinoma or Stage I and II breast carcinoma were reviewed retrospectively. At the time of the second opinion, patients completed a questionnaire regarding their initial surgical opinion and the reason for seeking consultation. RESULTS: Only 46% of patients had a complete discussion of treatment options prior to the second opinion. The second opinion changed management in 54 patients (20.3%). The most common finding was eligibility for BCT in patients who were offered only mastectomy. Definitive local therapy occurred at the second-opinion site in 65.8% of patients. The only predictors of remaining at the second-opinion site were insurance type (P = 0.008) and the patient's perception that options were not discussed at the initial opinion (P < 0.001). CONCLUSIONS: Second opinions provide useful information to patients and may change the management of their disease. They result in significant patient capture for an institution. PMID- 11920456 TI - Docetaxel plus epidoxorubicin as neoadjuvant treatment in patients with large operable or locally advanced carcinoma of the breast: a single-center, phase II study. AB - BACKGROUND: The objective of this study was to test the activity and toxicity of epirubicin plus docetaxel as primary chemotherapy for women with large, operable (T2; > 3 cm) or locally advanced (Stage III) breast carcinoma, including patients with inflammatory breast carcinoma. METHODS: In this single-center, open-label, single-stage, Phase II trial, epirubicin (75 mg/m(2); intravenous bolus) followed by docetaxel (80 mg/m(2); 1-hour intravenous infusion) was administered on Day 1 of each cycle for four cycles. RESULTS: Nine of 30 patients (30%) had inflammatory breast carcinoma. Twenty-three patients (76.7%; 95% confidence interval, 57.7-90.1) had a clinical objective response that was complete in 6 patients (20%). Twenty-seven patients (90%) underwent surgery that was conservative in 5 patients (16.7%). Pathologic response evaluation revealed four complete responses (13.3%; 95% confidence interval, 3.8-30.7). Grade 4 neutropenia was recorded in 80.0% of patients, and febrile neutropenia was recorded in one-third of patients. Anemia and thrombocytopenia were never severe. Other side effects were diarrhea (26.6%), oral mucositis (43.3%), and emesis (26.6%). CONCLUSIONS: Neoadjuvant chemotherapy with epirubicin plus docetaxel was a feasible treatment and was active in an unfavorable series of patients with locally advanced breast carcinoma, including patients with inflammatory breast carcinoma. PMID- 11920457 TI - Gemcitabine alone or with cisplatin for the treatment of patients with locally advanced and/or metastatic pancreatic carcinoma: a prospective, randomized phase III study of the Gruppo Oncologia dell'Italia Meridionale. AB - BACKGROUND: A prospective, randomized Phase III trial was performed to determine whether, compared with gemcitabine (GEM) alone, the addition of cisplatin (CDDP) to GEM was able to improve the time to disease progression and the clinical benefit rate in patients with advanced pancreatic adenocarcinoma. The objective response rate, overall survival rate, and toxicity patterns of patients in the two treatment arms were evaluated as secondary end points. METHODS: Patients with measurable, locally advanced and/or metastatic pancreatic adenocarcinoma were randomized to receive GEM (Arm A) or a combination of GEM and CDDP (Arm B). In Arm A, a dose of 1000 mg/m(2) GEM per week was administered for 7 consecutive weeks, and, after a 2-week rest, treatment was resumed on Days 1, 8, and 15 of a 28-day cycle for 2 cycles. In Arm B, CDDP was given at a dose of 25 mg/m(2) per week 1 hour before GEM at the same dose that was used in Arm A. On Day 22, only GEM was administered. Patients were restaged after the first 7 weeks of therapy and then again after the other 2 cycles. RESULTS: A total of 107 patients entered the trial: Fifty-four patients were randomized to Arm A, and 53 patients were randomized to Arm B. The median time to disease progression was 8 weeks in Arm A and 20 weeks in Arm B; this difference was statistically significant (P = 0.048). In Arm A, one complete response and four partial responses were recorded on the basis of an intent-to-treat analysis, with an overall response rate of 9.2% (95% confidence interval [95%CI], 3-20%). In Arm B, there were no complete responses, whereas 14 partial responses were achieved, with an overall response rate of 26.4% (95%CI, 15-40%). This difference in the overall response rates was statistically significant (P = 0.02). The tumor growth control rate (i.e., total number of patients who achieved complete responses, partial responses, and stable disease) was 42.6% (95%CI, 29-57%) in Arm A and 56.6% (95%CI, 42-70%) in Arm B. A clinical benefit was observed in 21 of 43 patients (49%) in Arm A and in 20 of 38 patients (52.6%) in Arm B without any significant difference. The median overall survival was 20 weeks for patients in Arm A and 30 weeks for patients in Arm B (P = 0.43). Toxicity was mild in both treatment arms, with no significant differences between the two groups except for the statistically higher incidence of Grade 1-2 asthenia in Arm B (P = 0.046). CONCLUSIONS: The addition of CDDP to GEM significantly improved the median time to disease progression and the overall response rate compared with GEM alone. The clinical benefit rate was similar in both arms, whereas the median overall survival rate was more favorable for Arm B, although the difference did not attain statistical significance. The authors conclude that the combination of CDDP and GEM currently may be considered as an optimal treatment for patients with locally advanced and/or metastatic adenocarcinoma of the pancreas. PMID- 11920458 TI - Evaluation of screening strategy for detecting hereditary nonpolyposis colorectal carcinoma. AB - BACKGROUND: The Amsterdam criteria are used worldwide for the clinical diagnosis of hereditary nonpolyposis colorectal carcinoma (HNPCC). In Japan, clinical criteria (JCC) have been proposed to identify as many HNPCC cases as possible, but the suitability of the JCC remains uncertain. In this article, the authors evaluate retrospectively whether the JCC are adequate to diagnose HNPCC compared with the Bethesda guidelines (BG) and also investigated useful screening methods for HNPCC. METHODS: The authors studied 452 colorectal carcinoma cases, of which 69 cases fulfilled the JCC (A, 12; B, 57) and 106 fulfilled the BG. Microsatellite instability (MSI) was examined for 452 cases. TGF beta RII, immunohistochemical staining, and germline mutations of hMLH1 and hMSH2 were analyzed in high-frequency MSI cases. RESULTS: High-frequency MSI was found in 21.7% (98 of 452). Germline mutations were detected in eight cases (hMLH1, three, hMSH2; five). Six cases fulfilled the JCC (A, four; B, two), and six fulfilled the BG. The germline mutation rate was significantly higher in the JCCA than in non-JCCA cases (33.3% vs. 0.91%; P < 0.001) and in cases with an age at onset younger than 50 years than older than 50 years (9.3% vs. 0.27%, P < 0.001). All germline mutation carriers had the TGF beta RII mutation. Immunohistochemically, a decreased nuclear staining was found in 57.3% (47 of 82) for hMLH1 and in 18.3% (15 of 82) for hMSH2. The frequency of predicted germline mutations was higher in cases with decreased hMSH2 than hMLH1 (33.3% vs. 6.4%; P = 0.016). CONCLUSIONS: The JCCA are suitable for selecting cases to analyze for gene mutations, but the JCCB are not useful for the clinical setting. The authors suggest that an age at onset younger than 50 years is also important for screening. Analyzing TGF beta RII mutations and immunohistochemical staining of hMLH1 or hMSH2 for cases with MSI phenotype are useful for selecting cases who should be tested for germline mutations. PMID- 11920459 TI - Comparison between positron emission tomography and computed tomography in the use of the assessment of esophageal carcinoma. AB - BACKGROUND: The role and potential value of positron emission tomography (PET) scanning in certain tumors has been widely investigated in recent years. The authors retrospectively assessed the performance of 18-F-fluorodeoxyglucose (FDG) PET in the assessment of esophageal squamous cell carcinoma (SCC). METHODS: The results using PET were compared with those using computed tomography (CT), and these were correlated with the pathologic findings. The authors studied 32 patients with thoracic esophageal SCC who had undergone radical esophagectomy. RESULTS: Uptake of FDG in the primary tumor was found in 25 of the 32 (78.1%) cases. Comparison of the FDG uptake and the clinicopathologic findings showed that there was a significant association between the FDG uptake and each of the depth of tumor invasion (P < 0.05), occurrence of lymph node metastasis (P < 0.01), and lymphatic invasion (P < 0.01). The survival rate in cases with high FDG uptake (standardized uptake value [SUV], >3) was significantly lower than that in cases with low FDG uptake (SUV, < 3; P < 0.05). In the evaluation of lymph node staging by the detection of lymph node metastasis, FDG-PET showed 77.8% sensitivity, 92.9% specificity, and 84.4% accuracy, and CT scanning showed 61.1% sensitivity, 71.4% specificity, and 65.6% accuracy. Positron emission tomography scanning showed a high degree of accuracy in the neck, upper thoracic, and abdominal regions. However, in the mid- and lower thoracic regions, the sensitivity was very low. The smallest lymph node metastasis that was detected by FDG-PET imaging was 6 mm. The average size of lymph node metastasis that was undetected by FDG-PET scanning was 7.3 mm (range, 1-17 mm). CONCLUSIONS: In conclusion, FDG-PET may be used as a noninvasive diagnostic technique in assessing the aggressiveness of the tumor and the prognosis in patients with esophageal SCC. During the preoperative diagnostic procedures, the sensitivity, specificity, and accuracy of lymph node staging is higher with FDG-PET than with CT imaging. In view of the high specificity of FDG-PET, it also gives useful information to guide the choice of treatment of esophageal carcinoma. PMID- 11920460 TI - Overexpression of caveolin-1 in esophageal squamous cell carcinoma correlates with lymph node metastasis and pathologic stage. AB - BACKGROUND: Caveolin-1 plays a regulatory role in several signaling pathways. Recently caveolin-1 was identified as a metastasis-related gene in prostate carcinoma. However, the clinical impact of caveolin-1 expression in esophageal carcinoma remains unknown. METHODS: One hundred thirty surgical specimens of esophageal squamous cell carcinoma were immunohistochemically assessed for caveolin-1 expression by polyclonal antibody. Caveolin-1 immunoreactivity was correlated with patients' clinicopathologic parameters and outcome. RESULTS: Positive caveolin-1 immunostaining was detected in 58 patients (44.6%). Positive caveolin-1 immunostaining correlated positively with pathologic stage (P = 0.029), pN (P = 0.023), and pM (P = 0.018). The overall survival rate was worse in patients with caveolin-1-positive tumors than in patients with caveolin-1 negative tumors (P = 0.0215). Univariate analyses identified caveolin-1 positivity (P = 0.0238), pT (P = 0.0002), pN (P < 0.0001), pM (P = 0.0002), lymphatic invasion (P = 0.0021), and positive surgical margin (P < 0.0001) as negative prognostic predictors. Multivariate analyses indicated that pT (P = 0.0296), pN (P = 0.0003), and a positive surgical margin (P = 0.0452) were independent prognostic factors. CONCLUSIONS: Over-expression of caveolin-1 is associated with lymph node metastasis and a worse prognosis after surgery in esophageal squamous cell carcinoma. PMID- 11920461 TI - Coexpression of ephrin-Bs and their receptors in colon carcinoma. AB - BACKGROUND: The erythropoietin-producing hepatoma amplified sequence (Eph) family is the largest subfamily of receptor tyrosine kinases (RTKs). The Ephs (receptors) bind to specific cell-bound ligands, called ephrins. The binding of this ligand-receptor system is dependent on cell-cell interactions. The ephrin Eph system is important in embryologic development and differentiation of the nervous and vascular systems. In the current study, the authors hypothesized that ephrins may play a role in the growth and development of colon carcinoma and may be expressed differentially in normal and malignant colonic tissues. METHODS: Reverse transcriptase-polymerase chain reaction (RT-PCR), Northern Blot analyses, and immunohistochemistry were used to examine 11 colon carcinoma cell lines and 20 human colon carcinoma specimens with adjacent uninvolved mucosa for the expression of EphB and ephrin-B family members. RESULTS: EphB2, EphB3, and EphB4 mRNA expression and ephrin-B2 mRNA expression was detected in all the cell lines and colon carcinoma specimens examined. Immunohistochemical analysis showed that ephrin-B2 had higher expression in the colon carcinoma specimens studied than in adjacent normal mucosa. Ephrin-B2 and EphB4 most frequently were expressed on the luminal surface of colon carcinoma epithelium. CONCLUSIONS: The results of the current suggest that the ephrin-Bs are expressed differentially in colon carcinoma and normal mucosa specimens and thus may play a role in the progression of colon carcinoma. Further studies are necessary to determine the functional role of ephrin-Bs in colon carcinoma angiogenesis and growth. PMID- 11920462 TI - Serum cross-linked carboxyterminal telopeptide of type I collagen (ICTP) as a prognostic tumor marker in patients with esophageal squamous cell carcinoma. AB - BACKGROUND: Serum cross-linked carboxyterminal telopeptide of Type I collagen (ICTP) is a metabolite of Type I collagen and has been reported to serve not only as a marker of bone metastasis but also as an indicator of treatment response and prognosis in several malignant tumors. The objective of this work was to evaluate the value of serum ICTP as a tumor marker in patients with esophageal squamous cell carcinoma (SCC). METHODS: In this study, pretreatment serum ICTP concentrations were measured by double-antibody radioimmunoassay in 50 patients undergoing tumor resection, chemoradiotherapy, or palliation for newly diagnosed esophageal SCC. The cutoff value for positive ICTP levels was defined as 4.50 ng/mL. As a comparison, serum concentrations of SCC antigen, carcinoembryonic antigen, cytokeratin-19 fragment, alkaline phosphatase, and lactate dehydrogenase were simultaneously evaluated. The results were categorized according to several clinicopathologic variables. RESULTS: Among the markers tested, ICTP showed the highest sensitivity (58.0%) in esophageal SCC sera. Positive ICTP levels were significantly correlated with tumor progression variables, such as tumor depth > or = T2, regional lymph node metastasis (N1), distant metastasis (M1), TNM stage > or = II, and maximal tumor length greater than 50 mm. Survival analyses of 29 patients who underwent curative resection demonstrated that patients who were positive for ICTP had significantly worse outcomes in terms of overall, disease specific, and disease free survival than those who were negative. Multivariate analyses confirmed that serum ICTP levels provided an independent prognostic indicator of overall and disease specific survival. However, serum ICTP values did not correlate with prognosis or treatment response in 17 unresectable cases treated by chemoradiotherapy. CONCLUSIONS: Our results indicated that serum ICTP concentrations might be a novel prognostic tumor marker for assessing the progression of esophageal SCC. PMID- 11920463 TI - Chemoprevention of gastrointestinal malignancies with nonsteroidal antiinflammatory drugs. AB - In multiple studies, the chronic use of nonsteroidal antiinflammatory drugs (NSAIDs) has been associated with a decreased incidence of several types of gastrointestinal (GI) neoplasia. This effect may be mediated by one of several intracellular mechanisms, some of which involve the inhibition of the cyclooxygenase-2 (COX-2) isoenzyme. In multiple studies of colorectal carcinoma, chronic NSAID use has shown a protective effect, with the majority of studies demonstrating a 30-70% risk reduction associated with NSAID use. The effect of NSAIDs on other types of GI neoplasia is less clear, but evidence suggests that chronic NSAID use may diminish the risk of esophageal and gastric carcinomas. Data assessing the effects of NSAIDs on the incidence of pancreatic and hepatic malignancies currently are too sparse and inconsistent to draw any conclusions. The newer COX-2 specific agents may provide a less GI-toxic alternative to nonselective NSAIDs as chemoprotective agents. PMID- 11920464 TI - Can initial prostate specific antigen determinations eliminate the need for bone scans in patients with newly diagnosed prostate carcinoma? A multicenter retrospective study in Japan. AB - BACKGROUND: The objective of the current study was to assess rigorously whether serum prostate specific antigen (PSA) determination can eliminate the need for bone scans in Japanese patients with newly diagnosed prostate carcinoma with serum PSA levels < or = 10 ng/mL. METHODS: A retrospective assessment of 1294 patients with newly diagnosed, untreated prostate carcinoma was conducted at the authors' institutions. All patients underwent a bone scan, serum PSA measurement, and core needle biopsy of the prostate. The receiver operating characteristic curve for identifying a positive bone scan based on serum PSA levels and a decision tree were analyzed to determine the expected 10-year cumulative cost and disease specific survival rate. Two competing strategies were used: PSA alone and PSA plus baseline bone scan. For the PSA-alone strategy, a baseline bone scan was performed only when the patient had a serum PSA level > 10 ng/mL. RESULTS: The proportion of positive bone scans in patients with serum PSA levels < or = 10.0 ng/mL was 1.33%. The area under the receiver operating characteristic curve was 0.870. Patients with a Gleason Grade > or = 3 tumors or with a Gleason score > or = 7 had a higher proportion of positive bone scans. The 10-year disease specific survival rates with the PSA-alone strategy and the PSA-plus-bone-scan strategy were the same. The PSA-alone strategy was minimally cost effective, with a savings of $16.00 (U.S.) in the cumulative net cost per patient over the PSA-plus bone-scan strategy. CONCLUSIONS: The current results suggest that baseline bone scans can be eliminated in patients with newly diagnosed prostate carcinoma in Japan who have serum PSA levels < or = 10 ng/mL. Apparently, it is possible to omit baseline bone scans for patients with a Gleason Grade < or = 2 tumors or with a Gleason score < or =6. PMID- 11920465 TI - Low expression of p27(Kip1) is associated with tumor size and poor prognosis in patients with renal cell carcinoma. AB - BACKGROUND: Proliferative activity in tumors depends on regulation of the cell cycle. p27(Kip1) (p27) plays a pivotal role as a negative regulator of the cell cycle. A decrease in p27 expression has been reported in many kinds of tumors, but little is known regarding p27 in patients with renal cell carcinoma (RCC). METHODS: Expression of p27 and the related cyclins (cyclin A, cyclin E, and cyclin D1) was examined immunohistochemically in 67 patients with of clear cell RCC. The Ki-67 labeling index (MIB-1 LI) and clinicopathologic parameters related to a poor prognosis also were analyzed. To determine their prognostic significance, univariate and multivariate survival analyses were performed. RESULTS: In tumors, there was considerable immunoreactivity for cyclin A, cyclin D1, and MIB-1, and the mean values for each were 1.08%, 16.1%, and 1.5%, respectively. Cyclin E expression was rare. The expression of p27 was correlated strongly with the expression of cyclin A (correlation coefficient, 0.432; P < 0.0004) and cyclin D1 (correlation coefficient, 0.476; P < 0.0004). Also, an inverse correlation was present between p27 expression and tumor size (P = 0.0377). In univariate analysis, the unfavorable prognostic factors were high TNM stage (P < 0.0001), large tumor size (P = 0.0016), high histologic grade (P = 0.0104), and low p27 expression (P < 0.0001). In multivariate analysis, high TNM stage (P = 0.0035) and low p27 expression (P = 0.0235) were independent prognostic factors for disease specific survival in patients with RCC. CONCLUSIONS: The results of this study suggest that low p27 expression may be a significant and independent, unfavorable prognostic factor in patients with renal cell carcinoma. PMID- 11920467 TI - Clinical utility of indium 111-capromab pendetide immunoscintigraphy in the detection of early, recurrent prostate carcinoma after radical prostatectomy. AB - BACKGROUND: Despite the ability of radical prostatectomy to eradicate prostate carcinoma, biochemical evidence of recurrent prostate carcinoma may be seen in approximately 40% of patients 15 years after they undergo surgery. Localization of recurrent disease after radical prostatectomy is difficult and may greatly influence subsequent clinical management. The authors examined the utility of indium 111 ((111)In)-capromab pendetide immunoscintigraphy to detect recurrent prostate carcinoma radiographically in men with early biochemical evidence of failure (serum prostate specific antigen [PSA] < or = 4.0 ng/mL) and assessed the minimum serum PSA level necessary for imaging recurrent disease. METHODS: Between May 1987 and August 1995, 255 hormone-naive men with a mean (+/- standard deviation) age of 65 years +/- 7 years who underwent radical prostatectomy for clinically localized prostate carcinoma were followed without adjuvant therapy until early PSA recurrence in this multicenter study. Preoperatively, all patients had negative bone scans and pathologically negative lymph nodes, and they did not undergo hormonal ablation, chemotherapy, or radiation therapy preoperatively or postoperatively until the (111)In-capromab pendetide scan was performed. All men in this study had postoperative serum PSA levels < or = 4.0 ng/mL at the time of radionuclide imaging. All men underwent imaging with the capromab pendetide scan to localize recurrent disease, and charts were reviewed to document clinical evidence of recurrence. RESULTS: Pathologic findings included mean Gleason scores of 6.7 +/- 1.2; pathologic tumors classified as pT2a (18%), pT2b (26%), pT3a (38%), pT3b (16%), and pT4a (2%); a pathologic lymph node status of pN0 (100%); positive surgical margins (44%); and perineural invasion (42%). Capromab pendetide uptake was seen in 72% of 255 men throughout a range of patients' postoperative serum PSA levels (0.1-4.0 ng/mL), with 31% of men having local uptake (prostatic fossa) only. Of 151 men who underwent additional imaging studies, 16 of 139 men (12%) and 15 of 92 men (16%) showed evidence of recurrent disease by bone scintigraphy and computed tomography scans, respectively. Gleason score, pathologic stage, perineural invasion, and margin status were not correlated significantly with the (111)In-capromab pendetide scan. CONCLUSIONS: Capromab pendetide imaging can localize early PSA recurrence and may guide appropriate treatment after patients undergo radical prostatectomy. No minimum serum PSA value was needed to potentially detect radiographic disease after surgery. Further confirmatory studies and long-term follow-up of this cohort documenting response to salvage therapy are needed to validate these imaging findings. PMID- 11920466 TI - HER-2 profiling and targeting in prostate carcinoma. AB - BACKGROUND: The clinical effects of targeting HER-2 in prostate carcinoma are not known. This study explored the feasibility of molecular profiling to determine the correlation between HER-2 expression, hormonal sensitivity, and the antitumor effects of trastuzumab and paclitaxel in patients with prostate carcinoma. METHODS: Patients with progressive androgen dependent (AD) and androgen independent (AI) prostate carcinoma were eligible to participate in the study. HER-2 expression was assessed on pretreatment tissue specimens, and patients were then assigned to one of four treatment groups: AD HER-2 positive, AD HER-2 negative, AI HER-2 positive, and AI HER-2 negative. They were treated with weekly trastuzumab at a dose of 2 mg/kg (after a 4 mg/kg loading dose) until they experienced disease progression, when weekly paclitaxel at 100 mg/m(2) was added. RESULTS: The authors screened 130 patients for HER-2 expression. In total, 23 patients were treated. Six eligible patients had HER-2 positive disease; therefore, only the AI HER-2 negative arm accrued to completion. All patients (100%) experienced disease progression on trastuzumab alone at or before the first 12 weeks of treatment. Fifteen patients received combined therapy: Seven patients (47%) experienced disease progression, 5 patients (33%) had stable disease, and 3 patients (20%) had a decline > or = 50% in prostate specific antigen PSA level or in soft tissue disease. HER-2 overexpression was found in significant proportions only in AI metastatic tissue samples (42% HER-2 positive; 95% confidence interval, 14-60%). In three of nine matched pairs, the AD prostate biopsy was HER-2 negative, and the AI metastatic sample was HER-2 positive. CONCLUSIONS: Trastuzumab is not effective as a single agent for the treatment of patients with AI HER-2 negative tumors. HER-2 expression varies by clinical state in patients with prostate carcinoma: Accurate HER-2 profiling requires sampling metastatic tissue in patients with metastatic disease. Further development of trastuzumab for the treatment of patients with metastatic prostate carcinoma is not feasible until more reliable and practical methods of sampling metastatic disease are developed to identify patients with HER-2 positive tumors. PMID- 11920468 TI - Cyclical change of hMSH2 protein expression in normal endometrium during the menstrual cycle and its overexpression in endometrial hyperplasia and sporadic endometrial carcinoma. AB - BACKGROUND: The role of hMSH2 protein, one of the major DNA repair proteins, until now, had not been elucidated in terms of normal endometrial function during the menstrual cycle. The current study was designed to address this issue and to determine whether the expression of hMSH2 is altered in the course of endometrial carcinogenesis. METHODS: Immunohistochemical reactivity with a monoclonal antibody against the hMSH2 protein was examined in endometrial tissue specimens from 45 patients with normal endometrium, 51 patients with endometrial hyperplasia, and 27 patients with endometrial carcinoma. Immunohistochemical expression of proliferating cell nuclear antigen (PCNA) also was studied in the same samples as a measure of the proliferative activity and was compared with hMSH2 expression in each sample. RESULTS: The functional layer of normal endometrium displayed cyclic changes in hMSH2 protein expression during the menstrual cycle, showing positive expression in the proliferative phase and becoming weak to negative in the secretory phase. This expression pattern was similar to that of PCNA. Sixty-three percent of endometrial carcinomas showed strong positivity for both hMSH2 and PCNA expression, and 7.4% had an intensity of hMSH2 protein expression similar to that found in normal proliferative endometrial glandular cells. There was only 1 sample (3.7%) that was completely negative for hMSH2 expression, and 26% of samples were weakly positive for PCNA and hMSH2 protein. All simple hyperplasias and the majority of complex and atypical hyperplasias showed positive immunoreactivity for hMSH2 and PCNA. CONCLUSIONS: This study demonstrates that hMSH2 protein expression changes during the menstrual cycle in parallel with proliferative activity. In most patients with sporadic endometrial carcinoma, the expression of hMSH2 protein is consistent with PCNA expression. In contrast, loss of hMSH2 expression is observed rarely in patients with sporadic endometrial carcinoma. PMID- 11920469 TI - Serum folate and homocysteine levels in head and neck squamous cell carcinoma. AB - BACKGROUND: Local and systemic metabolic alterations are always present in cancer. Carcinogenesis is associated with biochemical disorders, often nonspecific, that might promote or derive from tumoral progression. Thus, analysis of metabolic alterations may be a valuable approach to understanding the biochemistry of tumors and may provide a means of identifying new targets for therapy. The methionine cycle in particular has been extensively studied in human cancer. METHODS: The authors analyzed serum concentrations of two metabolites of such pathways, folate and homocysteine, in 42 patients affected by head and neck squamous cell carcinoma (HNSCC) in comparison with two control groups, composed of smokers and non smokers. RESULTS: Mean folate level was 5.8 +/- 2.1 ng/mL in carcinoma patients, 9.1 +/- 2.7 ng/mL in smoking controls, and 9.7 +/- 2.2 ng/mL in non smoking controls, with a statistically significant difference between carcinoma patients and smokers (mean difference: -3.3 ng/mL; 95% confidence interval [CI]: -4.234 to -2.366; P < 0.0001) and between carcinoma patients and non smokers (mean difference: -3.9 ng/mL; 95% CI: -4.67 to -3.13; P < 0.0001). Mean total homocysteine level was 10.4 +/- 5.3 microM in carcinoma patients, 7.8 +/- 2.5 microM in the non-smokers' group, and 8.3 +/- 2.8 microM in the smokers' group, with statistically significant differences between carcinoma patients and smoking controls (mean difference: 2.1 microM; 95% CI: 0.7056 to 3.494; P = 0.0034) and between carcinoma patients and non smoking controls (mean difference: 2.6 microM; 95% CI: 1.381 to 3.819; P < 0.0001). CONCLUSIONS: Differences in serum levels of folate and homocysteine might arise from tumor development and consequent metabolic alterations or might precede and promote tumor progression. If hypofolatemia is a risk factor for head and neck carcinogenesis, it might suggest a role for folate as a novel chemopreventive agent both in patients with precancerous lesions and in patients with treated HNSCC at risk for loco-regional recurrence and second primary tumors. PMID- 11920470 TI - Quality of life of nasopharyngeal carcinoma patients. AB - BACKGROUND: Quality of life (QOL) issues in patients with head and neck carcinoma are of importance beyond the incidence of these tumors because of the impact of the disease and its treatment on external appearance and function of the upper aerodigestive tract. Nasopharyngeal carcinoma (NPC) patients comprise a unique subgroup in whom, to our knowledge, QOL has not been studied directly. METHODS: Adult patients with NPC treated during the past 15 years at the Chaim Sheba Medical Center with a minimum follow-up of 6 months were included in the current study. Patients were mailed the revised University of Washington quality of life (UW-QOL) questionnaire and data pertaining to their disease were recorded. Patients with recurrent disease or another malignancy or those whose present status could not be ascertained were excluded from the study. QOL scores were analyzed based on treatment, disease stage, and patient age. RESULTS: Twenty eight patients of 35 disease-free patients (80%) responded to the questionnaire sent to patients meeting the study criteria. The mean score for general health was 3.1 (range, 1-5). Pain was not a significant factor. Other domains without noteworthy problems were speech and shoulder disability. The majority of patients described their appearance as normal or with minor changes, and questions concerning activity, recreation/entertainment, employment, and swallowing all scored > 70 (range, 0-100). Dry mouth, chewing, and ear problems were of major concern with the majority of patients and affected the QOL indices. Nevertheless, the overall mean QOL score of these patients was rated as "good" (4.2 on a scale of 1-6). CONCLUSIONS: In the current study, patients with NPC reported ear problems, difficulties in chewing, and dry mouth but their overall QOL appeared to be good. Ear problems such as secretory otitis media should be recognized at the time of presentation and treated. Conformal radiotherapy techniques sparing the salivary glands and temporal bone most likely will be useful in reducing the morbidity associated with treatment. PMID- 11920471 TI - Ossifying malignant mixed epithelial and stromal tumor of the liver: a case report of a previously undescribed tumor. AB - BACKGROUND: Malignant mixed tumors of the liver in adults are rare. To the authors' knowledge, twenty-four cases of primary malignant mixed hepatic tumors have been reported in the literature since the first description by Walter in 1896. Many of the previously reported cases are now considered mixed hepatoblastoma, primary hepatocellular carcinomas with focal sarcomatous metaplasia, or epithelioid hemangioendothelioma. METHODS: Clinical, radiologic, histologic and immunohistochemic methods were used to characterize an unusual example of a mixed malignant tumor of the adult liver that the authors descriptively refer to as "ossifying malignant mixed epithelial and stromal tumor" of the liver. RESULTS: In contrast to previously reported cases, the tumor in question was morphologically distinct. It was composed of three distinct neoplastic phenotypes: malignant spindle cells, with adenocarcinomatous differentiation, and extensive osteoid formation. The tumor was treated by partial hepatectomy. The patient was alive 8 years postoperatively as of December 2001. The distinctive combination of morphologic features and prolonged survival are much different from previously reported cases of mixed malignant hepatic tumors, which were typically rapidly fatal. CONCLUSIONS: The authors believe that the clinical and pathologic features of this patient's tumor are sufficiently distinctive to exclude it from the well-established categories of hepatic neoplasms. Histologically, the authors suggest the descriptive name "ossifying malignant mixed epithelial and stromal tumor of the liver" as a preliminary description for this tumor until the origin and relationship to other hepatic neoplasms are further characterized. PMID- 11920472 TI - Expression of cyclooxygenase-1 and cyclooxygenase-2 in bronchial epithelium and nonsmall cell lung carcinoma. AB - BACKGROUND: Cyclooxygenase (Cox) is the main target enzyme for the nonsteroidal antiinflammatory drugs that have been shown to suppress carcinogenesis in both experimental models and epidemiologic studies. METHODS: To evaluate its utility as an intermediate biomarker in bronchial chemoprevention trials, the authors examined Cox 1 and Cox 2 expression in normal and premalignant bronchial epithelial cells and nonsmall cell lung carcinoma (NSCLC) samples using an immunohistochemical staining technique. Included in the current study were 101 NSCLC samples and 77 bronchial biopsy samples obtained from 15 healthy smokers. RESULTS: In the normal bronchial epithelium, Cox 2 expression was found to be completely negative whereas Cox 1 expression was noted in a few scattered cells. The areas of basal cell hyperplasia and squamous metaplasia demonstrated the same pattern. There were relatively more Cox 2-positive tumors, as defined by positive staining in > 10% of tumor cells, than Cox 1-positive tumors (30 of 101 tumors [30%] vs. 14 of 101 tumors [14%]; P = 0.01). When tumor types were considered, there were more Cox 2-positive adenocarcinomas compared with squamous cell carcinomas (21 of 51 adenocarcinomas [41%] vs. 9 of 46 squamous cell carcinomas [20%]; P = 0.03). In contrast, fewer adenocarcinomas tended to show Cox 1 expression compared with squamous cell carcinomas (4 of 51 adenocarcinomas [8%] vs. 9 of 46 squamous cell carcinomas [20%]; P = 0.14). Although smokers tended to have more Cox 2-positive tumors than nonsmokers (29 of 91 tumors in the smokers [32%] vs. 1 of 10 tumors in the nonsmokers [10%]; P = 0.15), there was no statistically significant relation found between Cox 1 or Cox 2 expression and smoking status or prognostically significant clinicopathologic features. CONCLUSIONS: The results of the current study suggest that Cox 1 and Cox 2 expression may not be a useful intermediate biomarker in bronchial chemoprevention trials. Nevertheless, considering the patterns of Cox 1 and Cox 2 expression in tumor cells, Cox expression status may be a useful parameter when designing treatment strategies for a subset of NSCLC patients. PMID- 11920473 TI - Polymerase chain reaction on blood for the diagnosis of invasive pulmonary aspergillosis in cancer patients. AB - BACKGROUND: The premortem diagnosis of invasive pulmonary aspergillosis (IPA) is difficult to make and often missed. Several retrospective studies have suggested that Aspergillus polymerase chain reaction (PCR) performed on serum or whole blood is useful in diagnosing IPA. Two prospective studies confirmed this finding but included a small number of IPA cases. METHODS: The current study was performed to determine the diagnostic role of Aspergillus PCR performed on whole blood specimens from 54 patients with cancer and pulmonary infiltrates for which bronchoscopy with fungal stains and cultures were performed. PCR through amplified Aspergillus mitochondrial DNA and alkaline protease genes was performed on whole blood samples. RESULTS: The cohort in the current study was comprised of 4 patients with definite IPA, 7 patients with probable IPA, 7 patients with possible IPA, and 36 control patients with no evidence of IPA. The sensitivity, specificity, and positive and negative predictive values all were 100% for definite IPA cases and 57%, 100%, 100%, and 92%, respectively, for each of the probable and possible IPA cases. CONCLUSIONS: Aspergillus PCR on whole blood samples is highly sensitive for the detection of IPA and is predictive for IPA. The sensitivity appears to be correlated with the certainty of diagnosis as proven by tissue invasion. PMID- 11920474 TI - Different simian virus 40 genomic regions and sequences homologous with SV40 large T antigen in DNA of human brain and bone tumors and of leukocytes from blood donors. AB - BACKGROUND: Many studies found only a small fragment of the large T-antigen coding sequences in human tumors, raising doubts on authenticity of SV40 sequences detected in these samples. METHODS: Five different regions of SV40 DNA were investigated in 106 fresh human tumor biopsies (25 brain, 69 bone, 12 Wilms' tumors), 71 tumor-derived cell cultures (38 from brain and 33 from bone tumors) and normal tissues (5 fresh bone biopsies and 38 buffy coats) by polymerase chain reaction (PCR) techniques and filter hybridization with specific oligoprobes. Expression of SV40 Tag sequences was analyzed in human tumor specimens by RT-PCR. RESULTS: SV40 large T-antigen sequences were detected at high prevalence, in human biopsies of primary brain (37-44%) and bone (21-37%) tumors, in cell cultures derived from brain (30-54%) and bone (53-80%) tumors. SV40 Tag sequences were detected in 29% of buffy coats of blood donors. However, only four brain tumor cell lines showed all the five regions of the SV40 genome investigated. Expression of SV40 Tag sequences was found in 11 of 27 (41%) human tumor samples. DNA sequence analysis indicated that the PCR-amplified products belong to the SV40 wild type. Polymerase chain reaction products of Tag middle portion from 20 of 78 (26%) samples showed a 97% homology with telomeric sequences of human chromosomes 10 and 11. CONCLUSIONS: Authentic SV40 sequences were detected in human samples. The expression of SV40 Tag sequences indicates that SV40 could play a role, as a cofactor, in the onset/progression of specific human cancers. The inability to detect some regions of the virus genome may suggest that those regions are not required for tumor persistence or growth and have been lost or, alternatively, may be the result of assay conditions that were unable to PCR amplify those regions in the tumors. PMID- 11920475 TI - Prognostic factors in patients with thick cutaneous melanoma (> 4 mm). AB - BACKGROUND: The current study was conducted to examine the role of multiple clinical and histologic factors in the prognostic assessment of patients with thick primary melanoma (> 4 mm, classified as T4). METHODS: A retrospective analysis was performed in 329 patients with T4 cutaneous melanomas who were seen at the University of California at San Francisco Melanoma Center between 1978 and 2000. Fourteen histopathologic features were recorded prospectively by a single dermatopathologist. In addition, 9 clinical factors were analyzed. RESULTS: Several histologic factors were found to have a significant impact on the survival of patients with T4 melanoma. On univariate analysis, tumor thickness, ulceration, mitotic rate, microsatellites, vascular involvement, and histogenetic type of the primary tumor all were found to have a significant impact on the overall survival rate. Regional lymph node involvement reduced the overall survival of T4 patients dramatically. The 5-year overall survival of patients with lymph node-negative T4 disease was 61%, compared with 30% for those with lymph node-positive disease. When lymph node status was taken into account, tumor thickness, vascular involvement, and ulceration all remained independent predictors of overall survival on multivariate Cox regression analysis. Neither age, gender, nor anatomic location appeared to affect the recurrence-free or overall survival rates. CONCLUSIONS: Patients with thick primary cutaneous melanomas (> 4 mm) comprise a heterogeneous group. The presence or absence of lymph node involvement, vascular involvement, and ulceration appears to result in significantly divergent overall survival rates in this patient cohort. Consideration of these factors has important implications in the management of patients with T4 melanoma. PMID- 11920476 TI - Clinicopathologic correlates of solitary fibrous tumors. AB - BACKGROUND: Solitary fibrous tumors (SFTs) are rare fibrous neoplasms. Since their initial description as arising from the pleura, SFTs have been reported at a wide range of anatomic sites. To the authors's knowledge, there are no large series reporting both thoracic and extrathoracic SFTs nor are there any large series that analyze clinicopathologic correlates of tumor behavior. METHODS: Institutional soft tissue tumor and pathology databases were reviewed to identify patients. Pathologic material was reviewed by an experienced soft tissue pathologist and scored for the presence of a histologically malignant component. Clinical information was obtained from medical records and phone calls to patients. Statistical analysis was performed using the Student t test, Pearson chi-square test, and log-rank test. RESULTS: Seventy-nine patients with SFTs treated at a single institution over an 18-year period were identified. These tumors arose in a wide range of anatomic sites. Thoracic and extrathoracic SFTs had similar clinical and pathologic features, although extrathoracic tumors were more likely to be symptomatic on diagnosis. Seventy-five patients underwent surgical excision of a SFT at our institution. Overall, SFTs had a low rate of local recurrence and metastasis after surgical treatment. Extrathoracic SFTs had an increased risk of local recurrence that was small but statistically significant. There was no difference in metastasis-free survival between thoracic and extrathoracic SFTs. Positive surgical margins and the presence of a histologically malignant component were factors predicting worse local recurrence free survival. Positive surgical margins, tumor size greater than 10 cm, and the presence of a malignant component predicted worse metastasis-free survival. CONCLUSIONS: Solitary fibrous tumors are rare tumors that occur at all anatomic sites. Most SFT patients fare well after surgical treatment. Closer surveillance is warranted for those tumors that are larger than 10 cm or with the presence of a histologically malignant component. PMID- 11920477 TI - Osteosarcoma of the spine: experience of the Cooperative Osteosarcoma Study Group. AB - BACKGROUND: Due to the low incidence rate, the optimal strategy for the treatment of patients with spinal osteosarcoma is unknown. METHODS: Twenty-two patients with osteosarcoma of the spine (15 with tumors of the sacrum and 7 with tumors at other sites) who received chemotherapy according to the Cooperative Osteosarcoma Study Group protocol were analyzed. Six patients presented with metastasis, and 16 patients had no evidence of metastasis at the time of entry into the protocol. Of 12 patients who underwent excision of their tumors, 2 patients underwent wide excision, 3 patients underwent marginal excision, and 7 patients underwent intralesional excision. Eight patients received irradiation: Six patients received conventional radiotherapy only, one patient received neutron beam therapy, and one patient received samarium-153-ethylene diamine tetramethylene phosphonate therapy. Follow-up ranged between 24 months and 105 months (median, 47 months). RESULTS: The median survival was 23 months, and three patients have survived without disease for > 6 years. Patients with primary metastases (P = 0.004), large tumors (P = 0.010), and sacral tumors (P = 0.048) had lower overall survival compared with patients who had no metastasis, small tumors, and nonsacral tumors, respectively. There was a significant difference in overall survival between 5 patients who underwent either wide or marginal surgery and 17 patients who underwent either intralesional surgery or no surgery (P = 0.033). Among 17 patients who underwent no surgery or intralesional surgery, overall survival tended to be better in 7 patients who received irradiation compared with the overall survival in 10 patients who did not receive irradiation (P = 0.059). CONCLUSIONS: Patients with metastases, a large tumors, and sacral tumors had a poor prognosis in the current study with small numbers of patients. Wide or marginal excision of the tumor improved survival. Patients with osteosarcoma of the spine should be treated with a combination of chemotherapy and at least marginal excision for those with surgically accessible tumors. Postoperative radiotherapy may be beneficial. PMID- 11920478 TI - Immunohistochemical staining of GLUT1 in benign, borderline, and malignant ovarian epithelia. AB - BACKGROUND: Aberrant expression of the facilitative glucose transporter GLUT1 is found in a wide spectrum of epithelial malignancies. The authors describe an immunohistochemical study of GLUT1 expression in benign, borderline, and malignant ovarian epithelia. METHODS: One hundred forty one formalin-fixed, paraffin-embedded sections were immunostained with rabbit anti-GLUT1 using the streptavidin-biotin method. The samples were as follows: 3 endometriotic cysts, 9 serous cystadenomas, 15 mucinous cystadenomas, 17 noninvasive borderline implants, 3 invasive borderline implants, and 3 endosalpingiosis. In addition, 35 borderline tumors (26 serous, 7 mucinous, 2 seromucinous) and 56 adenocarcinomas (50 serous, 4 endometrioid, 2 mucinous) were stained. RESULTS: Benign serous and mucinous cystadenomas and endosalpingiosis were non-staining with GLUT1 antiserum. Twenty-eight of 35 borderline tumors (80%) stained positively, with weak to moderate (1-2+ out of 3) staining intensity and focal or patchy distribution. Seventeen noninvasive serous borderline implants were negatively stained; however, three invasive serous borderline implants were positively stained with GLUT1 antiserum. Fifty four of 56 ovarian carcinomas (96%) stained positively, with moderate to strong (2-3+ out of 3) intensity and multifocal distribution. CONCLUSIONS: GLUT1 is a consistent marker of ovarian epithelial malignancy. GLUT1 staining is absent in benign ovarian epithelial tumors, and shows progressively more staining in invasive tumors as compared to borderline tumors. Anti-GLUT1 antibody may be useful in distinguishing invasive from noninvasive serous borderline implants. PMID- 11920479 TI - Thymidine phosphorylase and vascular endothelial growth factor in patients with Stage I lung adenocarcinoma. AB - BACKGROUND: Thymidine phosphorylase (TP) has chemotactic activity in endothelial cells in vitro and angiogenic activity in vivo. However, the clinical significance of TP and cooperative roles of TP with other angiogenic factors have remained unclear in nonsmall cell lung carcinoma (NSCLC). METHODS: The authors stained for TP, vascular endothelial growth factor (VEGF), and basic fibroblast growth factor (bFGF) in 132 tumors from patients with Stage I NSCLC. They compared TP, VEGF, and bFGF expression levels with microvessel counts (MVCs), macrophage counts, mast cell counts, and clinical outcomes of patients with Stage I NSCLC. RESULTS: In adenocarcinoma samples, only stromal cell-TP expression and tumor cell-VEGF expression were associated with MVCs and mast cell counts but not with macrophage counts. In squamous cell carcinoma samples, there were no significant associations of the expression of any examined angiogenic factors with MVCs, mast cell counts, or macrophage counts. More importantly, only among patients with Stage I adenocarcinoma of the lung did patients in the stromal cell TP positive tumor group and in the tumor cell-VEGF positive tumor group have a significantly worse prognosis compared with patients in the stromal cell-TP negative tumor group and in the tumor cell VEGF negative group, respectively. In addition, among patients with Stage I adenocarcinoma, patients in the stromal cell-TP positive and tumor cell-VEGF positive tumor group had a significantly worse prognosis among the four groups. CONCLUSIONS: TP induction in tumoral stroma, but not in tumor cells, and tumor cell-VEGF induction may promote angiogenesis cooperatively in adenocarcinoma of the lung. Stromal cell-TP expression and tumor cell-VEGF expression may be important prognostic factors in adenocarcinoma of the lung, and stromal cell-TP expression may be a marker of active remodeling stroma in adenocarcinoma of the lung. PMID- 11920480 TI - Nonperioperative strokes in children with central nervous system tumors. AB - BACKGROUND: Nonperioperative strokes are rare yet potentially devastating events for children with central nervous system (CNS) tumors. The incidence of and risk factors for nonperioperative strokes in children with CNS tumors is unknown. METHODS: The authors performed a retrospective review of children from their institution with CNS tumors. The incidence of stroke in the nonperioperative period and the influence of patient demographic factors, coexisting genetic diseases, tumor type, and treatment modality on the subsequent occurrence of a stroke were determined. RESULTS: Eight hundred seven consecutive patients from the authors' institution with CNS tumors were observed for a combined 3224 nonperioperative years. Thirteen patients (1.6%) had a nonperioperative stroke, for an incidence of 4.03 strokes/1000 years of nonperioperative patient follow up. Eight patients were males, and the median age at diagnosis of a CNS tumor was 4.8 years (range, 0.3-18.6 years). The median duration from diagnosis of a CNS tumor until the occurrence of stroke was 2.3 years (range, 0.3-15.8 years). Among numerous potential risk factors individually examined by chi-square analysis, only treatment with radiation therapy was associated with the subsequent development of a stroke (chi-square, P = 0.007). By logistic regression analysis, treatment with radiation therapy and a diagnosis of an optic pathway glioma were the only statistically significant variables associated with a stroke. CONCLUSIONS: Strokes are much more common among children with CNS tumors. Children treated with radiation therapy and those with optic pathway gliomas have a higher association with the occurrence of a subsequent nonperioperative stroke. Because children with optic pathway gliomas may be at particularly high risk of stroke after radiation therapy, the desired beneficial therapeutic effects of irradiation must always be weighed against its potentially adverse effects, including stroke. PMID- 11920481 TI - Abnormalities of chromosome bands 15q13-15 in childhood acute lymphoblastic leukemia. AB - BACKGROUND: Recurring breakpoints in chromosome bands 15q13-15 occur infrequently in leukemia. To the authors' knowledge, the clinical significance of these breakpoints in childhood acute lymphoblastic leukemia (ALL) has not been previously investigated. METHODS: Centrally reviewed karyotypes of children with newly diagnosed ALL enrolled on Children's Cancer Group protocols from 1988 to 1995 formed the basis of the current report. Statistical analyses used chi-square tests for homogeneity of proportions, and outcome was analyzed using life table methods and associated statistics. RESULTS: Of 1946 cases with centrally reviewed and accepted cytogenetic analyses, 23 cases (1%) had breakpoints in chromosome bands 15q13-15. Most patients with 15q13-15 breakpoints had standard risk ALL, although breakpoints in 15q13-15 occurred more frequently in infants than in older children. The majority of these patients (16 patients; 70%) had balanced 15q13-15 rearrangements. Additional chromosomal abnormalities not involving 15q included abnormal 12p, abnormal 9p, Philadelphia chromosome, deletion 6q, and an 11q23 breakpoint. Thirteen (57%) 15q13-15 breakpoints occurred in pseudodiploid karyotypes; five (22%) were in hyperdiploid karyotypes with 47-50 chromosomes; two (9%) were in hyperdiploid karyotypes with > 50 chromosomes; and three (13%) were in hypodiploid karyotypes. Of the 23 patients with 15q13-15 breakpoints, 21 were survivors, 18 survived event-free for 2.2-9.3 years, and 3 were alive 1 to 3 years after a relapse at time of writing. CONCLUSIONS: The current study suggests that genes at 15q13-15 may be involved in the leukemogenesis of some cases of childhood ALL, but that with current intensive therapy such aberrations do not confer increased risk of treatment failure. PMID- 11920482 TI - Surgical view of the treatment of patients with hepatoblastoma: results from the first prospective trial of the International Society of Pediatric Oncology Liver Tumor Study Group. AB - BACKGROUND: Surgical resection is the cornerstone of treatment for patients with hepatoblastoma (HB). The Society of Pediatric Oncology Liver Tumor Study Group launched its first prospective trial (SIOPEL-1) with the intention to treat all patients with preoperative chemotherapy and delayed surgical resection. The objective of this article was to assess the assumed surgical advantages of primary chemotherapy. METHODS: Between 1990 and 1994, 154 patients age < 16 years with HB were registered on SIOPEL-1. The pretreatment extent of disease was assessed, and, after undergoing biopsy, patients were treated with cisplatin 80 mg/m(2) intravenously over 24 hours and doxorubicin 60 mg/m(2) intravenously over 48 hours by continuous infusion (PLADO). Generally, tumors were resected after four of a total of six courses of PLADO. RESULTS: One hundred twenty eight patients underwent surgical resection (13 patients underwent primary surgery, and 115 patients underwent delayed surgery after PLADO). A pretreatment surgical biopsy was performed in 96 of 128 patients (75%). Biopsy complications occurred in 7 of 96 patients (7%). Twenty-two patients showed pulmonary metastases at the time of diagnosis, and 7 patients underwent thoracotomy. Operative morbidity and mortality were 18% and 5%, respectively. Complete macroscopic surgical resection was achieved in 106 patients (92%), including 6 patients who underwent orthotopic liver transplantation. The actuarial 5-year event free survival (EFS) rate for all 154 patients in the study was 66%, and the overall survival (OS) rate was 75%. For the 115 patients who were included in the surgical analysis that followed the exact protocol, the EFS and OS rates were 75% and 85%, respectively. CONCLUSIONS: Biopsy is a safe procedure and should be performed routinely. Preoperative chemotherapy seems to make tumor resection easier. Reresection of a positive resection margin does not necessarily have to be performed, because postoperative chemotherapy showed good results. Resection of lung metastases can be curative if there is local control of the primary tumor; however, results showed that the patient's prognosis was worse. Surgical morbidity or mortality rates were not necessarily higher in large multicenter studies. More importantly, countries of lesser economic status also can contribute effectively to these trials. PMID- 11920483 TI - Importance of tumor regression assessment in predicting the outcome in patients with locally advanced rectal carcinoma who are treated with preoperative radiotherapy. AB - BACKGROUND: Locally advanced rectal carcinoma has a poor prognosis. However, since the introduction of preoperative radiotherapy, the outcome of patients with rectal carcinoma has been reported to have improved. Nevertheless, to the authors' knowledge few data are available regarding the histopathologic response to radiotherapy as assessed on surgical specimens as a potential predictive factor for outcome. METHODS: To estimate the effect of radiotherapy on rectal carcinoma, the authors retrospectively reviewed the surgical specimens of 102 patients with T3-4, N0 or > or = N1 rectal carcinoma and 1 patient with T2 but N1 rectal carcinoma. All patients were treated preoperatively with a hyperfractionated accelerated radiotherapy schedule in a prospective protocol (Trial 93-01). Using a standardized approach, tumor regression was graded using a system that varies from Grade 1 (tumor regression Grade [TRG] 1) when complete tumor regression is observed to Grade 5 (TRG5) when no tumor regression is observed. RESULTS: Radiotherapy resulted in tumor downstaging in 43% of the patients. There were 2 pT1 tumors (2%), 21 pT2 tumors (20%), 66 pT3 tumors (64%), and 14 pT4 tumors (14%) after treatment. Regional lymph nodes were involved in 55 patients (53%). None of the patients demonstrated a complete tumor regression after radiotherapy, but in 79% of the specimens a partial tumor regression was observed (TRG1: 0%; TRG2: 20%; TRG3: 39%; TRG4: 20%; and TRG5: 21%). The median actuarial overall survival (OS) and disease-free survival (DFS) were 52 months. Actuarial local recurrence rates at 2 years and 5 years were 6.4% and 7.6%, respectively. Univariate analysis showed the actuarial DFS to be significantly lower in patients with lymph node metastases (P = 0.0004) and advanced pT stages (pT3-4) (P = 0.03). A favorable outcome for OS, DFS, and local control was observed in patients with TRG2-4 (i.e., responders) compared with patients with TRG5 (i.e., nonresponders), but also in patients with low residual tumor cell density (TRG2, 3, and 4). On multivariate analysis, TRG remained an independent prognostic indicator for local tumor control. CONCLUSIONS: Tumor regression as well as residual tumor cell density were found to be predictive factors of survival in rectal carcinoma patients after preoperative radiotherapy. Even after preoperative radiotherapy, the pathologic stage of the surgical specimen remained a prognostic factor. The use of a standardized approach for pathologic evaluation must be implemented to allow comparison between the results of various treatment approaches. PMID- 11920484 TI - Combined chemotherapy and radiation therapy for head and neck malignancies: quality of life issues. AB - BACKGROUND: The standard of care for locally advanced head and neck carcinoma usually has been surgery followed by radiation therapy. Patient survival using this approach has been reported to be poor. The disfiguration resulting from surgery and the long-term morbidity of postoperative radiation often results in considerable distress. Concurrent chemotherapy and radiation was introduced to improve outcome. Excellent local control and survival results often have been reported in Phase II and Phase III studies. The acute toxicity of combined chemotherapy and radiation is significant. However, organ preservation may improve quality of life. This review article summarizes the findings from published series of surgery, postoperative radiation, radiation therapy alone, and chemoradiation with regard to quality of life issues for patients with locally advanced head and neck carcinoma. METHODS: A literature search was used to identify quality-of-life studies of postoperative radiation, radiation therapy alone, and chemoradiation in patients with locally advanced head and neck carcinoma. Factors affecting long-term quality-of-life issues in each treatment modality were identified, compared, and evaluated. RESULTS: Speech disorder, dysphagia, pain, and depression were found to be the common side effects affecting quality of life regardless of the treatment modality. Xerostomia is the major complication affecting patients undergoing radiation or chemoradiation. CONCLUSIONS: Acute side effects of combined chemotherapy and radiation therapy usually were found to resolve after treatment. Long-term morbidity is substantial because of xerostomia and severe dysphagia. However, preliminary studies suggest that because of organ preservation, patients may achieve a better quality of life after chemoradiation compared with the conventional use of surgery and postoperative radiation. PMID- 11920485 TI - Outpatient cancer drug costs: changes, drivers, and the future. AB - BACKGROUND: To the authors' knowledge, no analysis has examined the specific components of drug spending for overall cancer care. The authors' objective was to quantify and characterize trends in outpatient drug expenditures for cancer patients. METHODS: The authors retrospectively analyzed pharmacy and outpatient professional claims data from commercial and Medicare health maintenance organization enrollees with a solid tumor diagnosis in 1995 and 1998. Charges were subdivided by type of drug (antineoplastic drugs, chemotherapy adjuncts, supportive drugs, and drugs unrelated to cancer treatment). RESULTS: In 1995, 14,663 cancer patients received outpatient drug treatment and 13,829 patients in 1998. Total charges increased from $17.9 million (mean charge of $1218 per patient) to $27.9 million (mean charge of $2003 per patient), an average annual increase of 16%. Antineoplastic therapy constituted the largest component of cancer-related drug costs (67%) and represented 76% of the increase from 1995 to 1998. Most charges were incurred in the professional setting for agents administered by injection. The primary explanation for the increases appeared to be a shift in treatment patterns toward newer, more expensive antineoplastic agents. Supportive therapy represented 17% of the increase in cancer drug costs, followed by chemotherapy adjuncts (7%). Charges for drugs unrelated to cancer therapy increased by 21% per year. CONCLUSIONS: Antineoplastic therapy administered in an office or clinic was the single most important cost driver, with newer more expensive agents replacing older, less expensive drugs. Attempts to understand and control outpatient drug cost increases for cancer patients should focus primarily on antineoplastic therapy, especially the appropriate substitution of newer agents for older, less expensive alternatives. Some non chemotherapy cancer drugs may offer an opportunity to improve quality of life with a relatively small effect on overall cancer drug costs. PMID- 11920486 TI - Acupuncture for xerostomia: clinical update. AB - BACKGROUND: In the authors' clinic, patients with xerostomia after radiation therapy for malignancy have been offered acupuncture as potential palliation of their symptoms since November 1999. Preliminary data revealed that many patients achieve relief, even for symptoms refractory to pilocarpine therapy. METHODS: Acupuncture technique has been refined since the authors' previous publication. Originally described as a two-step process, a single treatment with eight needles is now used. Three points are treated in each ear, and one in the radial aspect of each index finger. Patients are also provided a sugar-free lozenge in the mouth to further stimulate salivation. Response is measured by the xerostomia inventory (XI). RESULTS: Fifty patients have undergone 318 treatments (median, 5; range, 2-15 treatments). Median follow-up since the first treatment is 224 days (range, 9-455 days). Median palliation as described by the XI was 9 points (range, 0-25 points). Response (defined as improvement of 10% or better over baseline XI values) occurred in 35 patients (70%). Twenty-four patients (48%) have received benefit of 10 points or greater on the XI. Duration of effect for 13 patients (26%) has exceeded 3 months. CONCLUSIONS: Acupuncture palliates xerostomia for many patients. A regimen of three to four weekly treatments followed by monthly sessions is now recommended, although some patients achieve lasting response without further therapy. PMID- 11920487 TI - Familial and second gastric carcinomas: a nationwide epidemiologic study from Sweden. AB - BACKGROUND: Familial risks in gastric carcinoma have been assessed mainly through case-control studies based on reported but not medically verified carcinomas in family members. Reliable data on familial risks are needed for prevention and clinical decisions. METHODS: The authors used the nationwide Swedish Family Cancer Database on 10.2 million individuals and more than 34,000 gastric carcinomas to calculate standardized incidence ratios (SIRs) and 95% confidence intervals (CIs) for gastric carcinoma in offspring, from birth to 66 years old, by carcinomas in family members. In addition, SIRs for second gastric carcinomas were analyzed. RESULTS: Standardized incidence ratios for gastric carcinoma were 1.31 (95% CI, 0.97-1.70) and 1.47 (95% CI, 1.08-1.92) when a parent presented with gastric carcinoma or gastric adenocarcinoma, respectively. The risk was 1.59 (95% CI, 1.10-2.16) in offspring whose diagnosis was at ages older than 50 years. Offspring risk from parental corpus carcinoma was of borderline significance whereas that from cardia carcinoma was below unity. The sibling risk for gastric carcinoma was 3.16 (95% CI, 1.35-5.72) and 5.75 (95% CI, 2.07-11.26) when diagnosed before age 50. The population attributable proportion of familial gastric carcinoma was 0.45%. Risks for second gastric carcinomas were increased in men and women after esophageal and skin carcinomas, and after non-Hodgkin lymphoma. CONCLUSIONS: The data suggest that environmental factors, perhaps Helicobacter pylori infections are the main reason for familial clustering of gastric carcinoma. The population attributable proportion of familial gastric carcinoma is much lower than that cited in the literature. The patterns of multiple carcinomas suggest that immunologic factors modulate susceptibility to gastric carcinoma. PMID- 11920488 TI - The specific role of isoflavones on estrogen metabolism in premenopausal women. AB - BACKGROUND: There is increasing evidence that dietary factors may play a role in the production, metabolism, and bioavailability of sex hormones and their impact on target tissues. The specific objective of this study was to evaluate the effectiveness of supplementing a group of premenopausal women who were free of breast carcinoma with a dietary supplement of isoflavones (40 mg per day) in producing a change in steroid hormones and menstrual cycle length. METHODS: Sixty eight consecutively recruited, premenopausal, omnivorous women of all races and ethnicities between the ages of 25 years and 55 years were admitted to the study and randomized to an experimental group supplemented with soy (40 mg genistein per day) or to a control group that consumed a placebo for a 12-week period. Changes in their anthropometric, nutritional, and hormonal biomarkers from early follicular phase were analyzed at baseline and post-intervention. RESULTS: Serum free estradiol and estrone levels decreased moderately in the experimental group. Serum hormone-binding globulin levels increased in 41.4% of women in the experimental group compared with 37.5% of women in the placebo group. Free estradiol decreased in 53.85% of women in the experimental group compared with 37.5% of women in the placebo group. Estrone decreased in 55.56% of women in the experimental group compared with 42.86% in the placebo group. Those women in the experimental group who were consuming soy had their mean menstrual cycle length increased by 3.52 days compared with a mean decrease of 0.06 days for women in the placebo group (P = 0.04) from baseline to the third menstrual cycle. In addition, women who were taking soy had their mean follicular phase increase by 1.46 days compared with a mean increase of 0.14 days for women who were taking the placebo (P = 0.08). CONCLUSIONS: These data suggest that increased isoflavone intake affects estrogen metabolism by altering the steroid hormone concentrations and menstrual cycle length, thereby demonstrating a potential to reduce the risk for breast carcinoma. PMID- 11920489 TI - Survival differences among Asian subpopulations in the United States after prostate, colorectal, breast, and cervical carcinomas. AB - BACKGROUND: Information is limited for Asian subgroups regarding survival after diagnosis of the common cancers amenable to routine screening. The authors examined survival after carcinomas of the prostate, colon/rectum, breast, and cervix separately for Chinese, Japanese, Filipinos, and non-Hispanic whites in the United States. METHODS: Using data from the Surveillance, Epidemiology, and End Results program, the authors compared the distributions of stage at diagnosis and computed 5-year cause specific survival probabilities, overall and by stage of disease, for cancer patients whose diagnosis was in 1988-1994 and who were observed through 1997. RESULTS: Among males, Filipinos were more likely to be diagnosed with advanced stage colorectal and prostate carcinomas than other Asians and non-Hispanic whites; they also experienced worse survival after these cancers. This survival deficit occurred across all stages of colorectal carcinoma and remained apparent within distant stage prostate carcinoma. Among females, Chinese were less likely to receive diagnoses of early stage colorectal carcinoma than Japanese and Filipinas. In addition, their survival was consistently lower across more advanced stages of disease. Chinese also experienced somewhat worse survival after diagnosis of early stage cervical carcinoma. Japanese were more likely to be diagnosed with early stage carcinomas but also tended to experience better survival after prostate, colorectal, and breast carcinomas regardless of stage. CONCLUSIONS: Chinese, Japanese, and Filipinos experienced unequal survival after these screenable carcinomas, indicating that certain groups may benefit from more aggressive screening efforts. The heterogeneity of cancer outcomes observed within the community classified as Asian reinforces the need for cancer statistics to be reported for disaggregated subgroups. PMID- 11920492 TI - Pathologic analysis of tumor size and lymph node status in multifocal/multicentric breast carcinoma. AB - BACKGROUND: For unifocal invasive breast carcinoma, increasing tumor diameter predictably correlates with a greater frequency of lymph node involvement, thereby facilitating treatment decisions. In invasive breast tumors presenting with multiple nodules, however, it is unclear whether tumor size correlates with lymph node dissemination in a similar manner. METHODS: The authors analyzed a series of 101 invasive breast carcinomas presenting with multiple macroscopically apparent lesions (2 foci: n = 77; 3: n = 20; 4: n = 4). Two different assessments of the tumor size (diameter of largest focus and combined diameter of all the foci) were then correlated with the status of axillary lymph nodes. For comparison with unifocal tumors, the authors used both external and internal control series (the latter consisting of 469 patients from their institution). The associations between lymph node status, tumor size, and multifocality were modeled using univariate and multivariate logistic regression, for each modality of tumor size assessment. RESULTS: The logistic curves for multifocal and unifocal tumors were significantly different when the largest diameter was used as a tumor size estimate. Multifocal cases had higher frequencies of lymph node involvement than unifocal lesions of similar size category. In a multivariate logistic regression, the odds ratio of positive lymph node status in multifocal versus unifocal cases was 2.8 using largest diameter as a tumor size estimate (P < 0.0001). When the combined diameter assessment was used, however, the regression curve of multifocal cases was similar to that of unifocal cases, and the frequency of lymph node positivity was not significantly different in multifocal versus unifocal cases of the same size (odds ratio, 1.4; P = 0.13). CONCLUSIONS: The authors' results show that, if aggregate diameters are used, unifocal and multifocal breast carcinomas are similar with respect to frequency of regional lymph node metastasis. Currently used algorithms, which use the diameter of the largest nodule, result in understaging of multifocal breast carcinomas due to underestimation of actual tumor size. PMID- 11920490 TI - Cancer trends among Hispanic men in South Florida, 1981-1998. AB - BACKGROUND: Hispanics now represent a majority of residents in Miami-Dade County, Florida. In this report, the authors present new cancer incidence and mortality data for South Florida's Hispanic men for the period 1990-1998 and compare them with data from a previous report from the 1980s. Periodic updating of cancer incidence data, reflecting current population distribution, lifestyle, and environmental risk factors, is necessary to inform cancer prevention and control activities optimally. METHODS: The study population consisted of all incidents of cancer (1981-1998) occurring in males from Miami-Dade County, as determined from the Florida Cancer Data System data base; patients were divided into two 9-year periods for analysis. Age-standardized incidence and mortality rates were computed for 14 common cancer sites, and rates for Hispanic men were compared with the rates for non-Hispanic white men as standardized rate ratios (SRRs) with 95% confidence intervals (95% CIs). Incidence and mortality trends were determined using linear regression analysis. RESULTS: Nearly 70,000 incident cancer cases were analyzed. For 1990-1998, the top five incident cancers for both race/ethnic groups were the same. The overall decreased cancer risk for Hispanic men (SRR, 0.80; 95% CI, 0.79-0.82), compared with non-Hispanic white men, remained essentially constant over the two study periods. Cancer incidence increased similarly for the two race-ethnic groups; cancer mortality decreased, with a sharper decrease for non-Hispanic white men, resulting in apparent convergence of mortality trends recently. CONCLUSIONS: Differences in cancer risk for South Florida's Hispanic men have not attenuated over the past 20 years. With cancer incidence significantly less for Hispanic men, their mortality rate approaches that of non-Hispanic white men, and cancer prevention and control strategies targeted for this ethnic group become increasingly important. PMID- 11920493 TI - Breast carcinoma treatment received by women with disabilities compared with women without disabilities. AB - BACKGROUND: Disability may make it difficult to lie flat or abduct the arm to deliver radiation therapy, imposing a high risk for radiation-induced side effects or difficulty in positioning patients for mammography. The goal of the current study was to determine the differences in treatment options experienced by women with physical disabilities compared with those without disabilities. METHODS: Chart review of 234 women who underwent surgery for breast carcinoma between June and September 1998 in a national comprehensive cancer center was conducted. Thirty-nine of the women had physical disabilities; the remaining 195 women were without disabilities. Hierarchical logistic regression was used to determine whether women with disabilities were less likely than women without disabilities to be treated with breast-conservation surgery (BCS) or neoadjuvant chemotherapy. RESULTS: Women with disabilities underwent BCS 38% of the time, whereas women without disabilities underwent BCS 49% of the time. Neither the presence nor absence of disability (P = 0.146) nor patient age (P = 0.747) were found to be significant predictors of BCS. However, the disease stage at the time of the surgery was reported to be a significant predictor of BCS (P = 0.007). The group of patients with disabilities received neoadjuvant chemotherapy 13% of the time, whereas women without disabilities received it 29% of the time. Disability was not found to be a significant predictor of whether a patient received neoadjuvant chemotherapy. Using hierarchical logistic regression, patient age was found to be a significant predictor of neoadjuvant chemotherapy before the disease stage was entered into the regression equation. There were no data to support the hypothesis that breast carcinoma is at an advanced stage when diagnosed in women with disabilities. CONCLUSIONS: These findings are clinically significant in that they indicate that women with disabilities are less likely to undergo BCS and are less likely to receive neoadjuvant chemotherapy compared with women without disabilities, but the difference did not reach statistical significance. To the authors' knowledge, there are no data to support the hypothesis that disabled women are diagnosed at a more advanced stage of disease compared with women without disabilities. PMID- 11920494 TI - ErbB2 expression is correlated with increased survival of patients with osteosarcoma. AB - BACKGROUND: Elevated ErbB2 expression and gene amplification have been shown to be associated with poor prognosis in many cancers. Recently, it has been demonstrated that overexpression of ErbB2 protein in osteosarcoma is associated with the presence of pulmonary metastasis and decreased survival. By contrast, a previous study showed that the expression of ErbB2 declines in individual osteosarcomas as they become metastatic. In the current study, the authors determined the relation between ErbB2 status and outcome in a large number of selected patients with high-grade osteosarcoma. METHODS: ErbB2 status was determined immunohistochemically in biopsy specimens of osteosarcoma of the extremities from 81 patients who were treated with surgery and chemotherapy. None of the patients had metastatic disease at presentation (Stage II), and all were followed-up for at least five years. The ErbB2 status was analyzed in relation to the lengths of event-free and overall survival. RESULTS: Of the 81 tumors examined, 51 (61%) demonstrated high levels of ErbB2 expression. The presence of increased levels of ErbB2 in osteosarcoma was significantly associated with the increased probability of event-free (72.2% v. 45.6% at 5 years, P = 0.03) and overall survival (79.7% v. 58.2% at 5 years, P = 0.03). Cox multivariate analysis showed that the risk of adverse events and death was increased substantially (rate ratio: 2.24 and 2.54; 95% confidence interval, 1.07-4.72 and 1.09-5.67, respectively) among patients with decreased levels of ErbB2 protein in tumor cells, as compared with patients who had increased levels of ErbB2 in tumor cells. CONCLUSIONS: In patients with high-grade osteosarcoma without metastatic disease at presentation and treated with surgery and chemotherapy, the presence of increased levels of ErbB2 in tumor cells is associated with a significantly increased probability of event-free and overall survival. Further data are needed before this marker can be used in making clinical decisions. PMID- 11920495 TI - Postoperative radiotherapy for patients with completely resected thymoma: a multi institutional, retrospective review of 103 patients. AB - BACKGROUND: Optimal management of postoperative radiotherapy for patients with completely resected thymoma remains controversial. This study was conducted to assess the efficacy of postoperative mediastinal irradiation in patients with completely resected thymoma. METHODS: The records of 103 patients with completely resected thymoma who received postoperative mediastinal irradiation during the period between 1979 and 1998 were reviewed. The distribution according to Masaoka stage was Stage I in 17 patients, Stage II in 61 patients, and Stage III in 25 patients. Fifty-two patients were treated with involved field (IF) irradiation, and 51 patients were treated with irradiation of the whole mediastinal field with or without boost (WM irradiation). The total radiation dose to the primary tumor bed was 30-61 grays (Gy), with a median dose of 40 Gy. No patients received chemotherapy during the initial treatment. The median follow-up of the 82 living patients was 112 months (range, 24-244 months). RESULTS: The 10-year actuarial overall and disease free survival rates for all patients were 81% and 79%, respectively. The 10-year actuarial overall survival rate was 100% for patients with Stage I disease, 90% for patients with Stage II disease, and 48% for patients with Stage III disease. In the analysis, clinical stage alone had a statistically significant impact on both overall survival and disease free survival (P < 0.0001 for both). Recurrent disease was observed in 17 patients, and the pleura was the most frequent site of first recurrence. Of 12 patients who had pleural recurrences, 11 patients had pleural dissemination remote from the initial tumor site. No recurrence was observed in any of the 17 patients with Stage I disease, and 6 of 61 patients (10%) with Stage II disease and 11 of 25 patients (44%) with Stage III disease experienced recurrences. With regard to intrathoracic recurrences, there were no recurrences within the irradiated field in any of the 103 patients, and no dose response correlation was seen in intrathoracic control (incidence of intrathoracic recurrence: 2 of 19 patients in the group that received < 40 Gy, 6 of 45 patients in the group that received 40 Gy, and 7 of 39 patients in the group that received > 40 Gy). With respect to treatment field, mediastinal recurrences were observed in 4 of 52 patients (8%) who were treated with IF irradiation, whereas 0 of 51 patients who were treated with WM irradiation experienced mediastinal recurrences. Pleural-based recurrences were observed both in patients who were treated with IF irradiation (7 of 52 patients) and in patients who were treated with WM irradiation (5 of 51 patients). According to the degree of pathologic tumor invasion, 0 of 71 patients without pleural invasion had pleural-based recurrences (0 of 17 patients with Stage I disease, 0 of 51 patients with Stage II disease, and 0 of 3 patients with Stage III disease), whereas 12 of 32 patients (38%) with pleural invasion had pleural-based recurrences (4 of 10 patients with Stage II disease and 8 of 22 patients with Stage III disease). CONCLUSIONS: The current results indicated that WM irradiation with a total dose of 40 Gy was effective in preventing mediastinal recurrence for patients with completely resected thymoma. However, in patients with pathologic pleural invasion of the tumor, mediastinal irradiation alone was insufficient to avoid pleural-based recurrence. PMID- 11920496 TI - Somatostatin analogs and prednisone in advanced refractory thymic tumors. AB - BACKGROUND: Therapeutic options to cure advanced, recurrent, and metastatic thymic tumors are limited. Evidence of a high uptake of indium-labeled octreotide ((111)In-DTPA-D-Phe(1)-octreotide) in thymic tumors and the curative application of somatostatin analogs and prednisone in one patient with thymoma and pure red cell aplasia led the authors to start a Phase II study. METHODS: Sixteen patients with advanced thymic tumors, unresponsive to conventional chemotherapeutic regimens, were enrolled in the study. The schedule includes administration of somatostatin analog octreotide (1.5 mg/day subcutaneously) associated with prednisone (0.6 mg/kg/day orally for 3 months, 0.2 mg/kg/day orally during follow up). In 8 cases, octreotide was replaced by the long-acting analog lanreotide (30 mg/every 14 days intramuscolarly). Treatment was prolonged until progression of disease was documented. Overall response rate, survival, progression free survival, and toxicity were evaluated. RESULTS: The overall response rate among 16 evaluable patients was 37%. One patient (6%) had a complete response, 5 (31%) had a partial response, 6 obtained a stabilization of disease, and 4 progressed during the treatment. After a median follow-up of 43 months, the median survival was 15 months, and median time to progression was 14 months. Treatment was generally well tolerated with acceptable toxicity: cholelithiasis (1 patient), Grade 2 cushingoid appearance (3 patients), Grade 1 diarrhea (5 patients), Grade 2 hyperglycemia (3 patients). CONCLUSIONS: Treatment with somatostatin analogs and prednisone has shown efficacy in patients with recurrent and metastatic malignant thymic tumors refractory to standard therapeutic options. The results obtained are very satisfactory given the lack of effective alternative treatments. Such therapy is not burdened by the same toxicity of chemotherapy; thus, it can be administered to heavily pretreated patients. Somatostatin analogs and prednisone are well tolerated, and the long-acting analog lanreotide, which requires fewer injections, improves patients' compliance. PMID- 11920497 TI - The APC/beta-catenin pathway in ulcerative colitis-related colorectal carcinomas: a mutational analysis. AB - BACKGROUND: Although the APC/beta-catenin pathway is known to play a crucial role in sporadic colorectal carcinogenesis, its influence on ulcerative colitis (UC) related neoplastic progression is unknown. To elucidate the role of the APC-/beta catenin pathway in UC-related carcinogenesis, the authors identified APC and beta catenin mutations in a set of UC-related and sporadic colorectal carcinomas. METHODS: The mutational cluster region of APC (codon 1267 to 1529) and exon 3 of the beta-catenin were directly sequenced. RESULTS: Only 1 of 30 UC-related tumors (3%) showed an APC mutation whereas 11 of the 42 sporadic carcinomas (26%) had mutations within the mutational cluster region. Within the sporadic carcinoma group, only 8% of the right-sided carcinomas showed APC mutations whereas 50% of the left-sided carcinomas had mutations within the mutational cluster region. None of the tumors in either group showed a beta-catenin mutation. CONCLUSIONS: Mutations of the APC and beta-catenin are rare in UC-related tumors. These genes may be altered because of mutations outside the regions studied, or by epigenetic silencing. Alternatively, other proteins involved in the APC/beta-catenin signaling cascade may be altered, or this pathway may be involved infrequently in UC-related carcinogenesis. The significant difference in frequency of APC mutations between right- and left-sided sporadic tumors suggests different molecular pathways in these two tumor sites. PMID- 11920498 TI - Patterns of gastric atrophy in intestinal type gastric carcinoma. AB - BACKGROUND: Multifocal atrophic gastritis (MAG) is currently considered a precancerous lesion leading to intestinal type gastric carcinoma. The current study aimed to describe the topography of atrophy in stomachs with early gastric carcinoma. METHODS: Resected stomachs from patients with intestinal type gastric carcinoma were routinely processed, sectioned (an average of 108 sections/stomach), and stained with a triple stain. Sections were scored on a visual analog scale for Helicobacter pylori and intestinal metaplasia. The type of epithelium (antral, oxyntic, transitional) was recorded. Atrophy was defined as the loss of normal glandular components and included intestinal metaplasia and/or pseudo-pyloric metaplasia of the corpus. Pseudo-pyloric metaplasia was identified by the presence of pepsinogen I in mucosa that was topographically corpus but phenotypically antrum. RESULTS: Sixteen stomachs with intestinal type gastric carcinoma were examined. In none of the specimens examined was MAG (independent foci of atrophy) identified. In the majority (88%), atrophy was present as a continuous sheet. Islands of intestinal metaplasia (multifocal intestinal metaplasia) were present within a sheet of pseudo-pyloric metaplasia. A few specimens (12%) had a non-atrophic corpus with almost total replacement of antral epithelium with intestinal metaplasia. Multifocal dysplasia distant from the original tumor was found both in areas with and without intestinal metaplasia. CONCLUSIONS: Contrary to popular belief, atrophy in intestinal type gastric carcinoma is not present as independent foci, but rather as a continuous sheet. Previous studies failed to identify pseudo-pylori metaplasia as a marker for atrophy. PMID- 11920499 TI - CD3-zetachain expression of intratumoral lymphocytes is closely related to survival in gastric carcinoma patients. AB - BACKGROUND: Impaired or reduced CD3 zeta chain (CD3-zeta) expression in T cells has been identified in various cancers and may be associated with an ineffective immune response. The clinical significance of CD3-zeta chain expression in tumor infiltrating lymphocytes (TILs) in gastric carcinoma remains unclear. METHODS: The authors immunohistochemically investigated CD3-zeta expression in TILs in 185 patients who had undergone curative gastrectomy. CD3-zeta/CD3 epsilon (CD3 epsilon) ratios were calculated. Patients were divided into two groups: a normal CD3-zeta group (n = 121) and a reduced CD3-zeta group (n = 64). Patients with a zeta per epsilon ratio of greater than 66% were placed in the normal CD3-zeta group. RESULTS: Patients in the normal CD3-zeta group had fewer lymph node metastasis (P < 0.01) and a shallower depth of invasion (P < 0.05) than those in the reduced CD3-zeta group. The 5-year survival rate was 72% in the normal CD3 zeta group, which was significantly better than that in the reduced CD3-zeta group (55%; P < 0.01). When stratified according to clinical stage, the prognostic value was significantly different only in Stage IV patients. Multivariate analysis showed that CD3-zeta expression was an independent prognostic factor (P = 0.03) next to depth of invasion and lymph node involvement. CONCLUSIONS: Reduced CD3-zeta expression in TILs was strongly correlated with progressive disease in gastric carcinomas. CD3-zeta expression in TILs is considered an immunologic, independent prognostic marker in gastric carcinoma patients. CD3-zeta normalization with cytokine treatment may lead to prolonged survival in advanced gastric carcinoma patients. PMID- 11920500 TI - Association of the -160 C --> a promoter polymorphism of E-cadherin gene with gastric carcinoma risk. AB - BACKGROUND: A -160 C --> A polymorphism in the promoter region of E-cadherin has been shown to decrease gene transcription. This allelic variation might be a potential genetic marker for identifying individuals at risk for cancer. There remains no report regarding the polymorphism of E-cadherin in gastric carcinoma (GC). METHODS: A hospital-based case-control study, including 201 GC cases and 196 unaffected controls, was performed. DNA from peripheral blood samples was examined by polymerase chain reaction-restriction fragment length polymorphism. Logistic regression analyses were used to compute odds ratio (OR) and 95% confidence interval (CI) after adjusted for Helicobacter pylori infection, smoking, and other relevant factors. RESULTS: In 196 healthy Taiwanese, the distribution of genotype C/C was 42.3%, C/A was 48.0%, and A/A was 9.7%. The frequency of variant A/A genotype in GC case (4 of 201, 2%) was significantly lower than that of controls (19 of 196, 9.7%) (P < 0.005), conferring a 5-fold decrease in the risk of GC (OR, 0.20; 95% CI, 0.06-0.56) compared with the C/C genotype. Stratification of the GC cases according to their location (cardia and noncardia), histology (intestinal and diffuse), tumor stage (early and advanced), and lymph node metastasis (positive and negative) failed to reveal any heterogeneity with respect to E-cadherin genotype. CONCLUSIONS: The authors' data suggest that individuals with E-cadherin -160 A/A genotype have a decreased risk of GC. Further work is mandatory to clarify the functional relevance of the A allele in vivo and to confirm the inverse association of the A/A genotype with GC in large epidemiologic studies. PMID- 11920501 TI - Macrophage migration inhibitory factor evaluation compared with prostate specific antigen as a biomarker in patients with prostate carcinoma. AB - BACKGROUND: Cytokines are polypeptides that constitute a class of chemical mediator molecules that modulate cell growth by inducing specific target gene expression. The objective of this study was to evaluate the clinical usefulness of serum evaluation of the cytokine macrophage migration inhibitory factor (MIF) in patients undergoing routine prostate specific antigen (PSA) screening. METHODS: In this preliminary, retrospective study, the authors report the development of an enzyme-linked immunosorbent assay (ELISA) for MIF determination in serum samples. A polymerase chain reaction (PCR)-based assay investigated associations between MIF expression and prostate carcinoma (CaP). The authors developed a relative quantitative reverse transcriptase-PCR assay to determine MIF mRNA amounts within laser-capture microscopy (LCM)-dissected prostate epithelial cells. RESULTS: A comparison of serum MIF levels and total PSA levels identified a positive correlation (correlation coefficient [r2] = 0.61; P < 0.001; n = 509 patients), suggesting an association between elevated serum concentrations of these proteins and CaP. A correlation of serum MIF levels with a diagnosis of CaP demonstrated that patients with a previous CaP diagnosis had significantly elevated serum MIF concentrations (mean +/- standard deviation, 6.8 +/- 0.87 ng/mL; P < 0.001). To associate altered serum MIF levels with MIF mRNA expression within prostate epithelial cells, LCM-dissected prostate epithelial cells (formalin fixed biopsies from three different patients) were used to determine MIF mRNA amounts by PCR analysis. On average, MIF mRNA amounts were 6.5 times higher in CaP epithelial cells that were invasive to the margin compared with MIF mRNA amounts in normal prostate epithelial cells within the same biopsy specimen. CONCLUSIONS: The ELISA data from the current study suggested an association between increased MIF expression and CaP and suggested that serum MIF concentration may serve as a prognostic marker for CaP. PMID- 11920502 TI - Phase II evaluation of docetaxel plus one-day oral estramustine phosphate in the treatment of patients with androgen independent prostate carcinoma. AB - BACKGROUND: Recent clinical trials have shown antitumor activity with the combination of docetaxel plus estramustine phosphate (EMP) in the treatment of patients with androgen independent prostate carcinoma (AIPC). However, the most commonly employed treatment schedules with EMP have been associated with significant gastrointestinal, cardiovascular, and thromboembolic toxicity. The authors hypothesized that the therapeutic index of the combination of docetaxel plus EMP for patients with prostate carcinoma could be enhanced by reducing the incidence and severity of EMP-associated toxicity, which could be accomplished by shortening the duration of exposure to EMP. To preserve the therapeutic synergism between docetaxel and EMP, they designed a regimen employing higher doses of oral EMP administered on the day of the docetaxel infusion. METHODS: From June 1, 1998 through September 28, 2000, 42 patients with AIPC were registered to receive docetaxel (70 mg/m2 intravenously over 1 hour) and EMP (280 mg orally every 6 hours x 5 doses) every 21 days, up to a maximum of 6 cycles. Dexamethasone was administered prior to docetaxel and coumadin 2 mg orally every day was taken during the study treatment period. Patient characteristics included a median age of 68 years, a median Eastern Cooperative Oncology Group performance status of 1, a median prostate specific antigen (PSA) level at study entry of 110.5 ng/mL, and a median of 2 prior hormonal manipulations. Ten patients (25%) had received prior chemotherapy, and 14 patients (33%) had received prior palliative radiation therapy. RESULTS: Forty patients were evaluable for response and toxicity. Eighteen patients (45%; 95% confidence interval, 29-62%) had a decline > 50% in PSA level that lasted > 4 weeks with a median time to PSA progression and a median duration of PSA response of approximately 4.0 months. Four of 20 patients (20%) had partial soft tissue responses. Ten of 17 symptomatic patients (59%) had improvement in pain. The median survival for all patients was 13.5 months. The most prominent Grade 3 and 4 toxicities were reversible myelosuppression and fatigue. Nausea, emesis, diarrhea, and peripheral edema were minimal. No thromboembolic or hepatic complications were seen. CONCLUSIONS: Docetaxel plus 1 multidose day of oral EMP was active in patients with AIPC and was associated with an acceptable toxicity profile. Overall, the therapeutic index of this regimen compared favorably with regimens that employed a longer administration of EMP. PMID- 11920503 TI - Presence of active gelatinases in endometrial carcinoma and correlation of matrix metalloproteinase expression with increasing tumor grade and invasion. AB - BACKGROUND: The actions of the extracellular-matrix degrading enzymes, matrix metalloproteinases (MMPs), are implicated in tumorigenesis. The cellular localization of MMP-2, MMP-9, membrane type 1 (MT1)-MMP, tissue inhibitors of metalloproteinases (TIMPs) 1-3, and the presence of active gelatinases were investigated in endometrial carcinoma. METHODS: Endometrial carcinomas were grouped according to histologic grade (Grades 1-3), depth of myometrial invasion (0, < 50%, > 50%) and the presence of vascular/lymphatic invasion. Twenty-nine endometrial carcinoma biopsies were investigated immunohistochemically to determine the tissue localization of MMP-2 (gelatinase A), MMP-9 (gelatinase B), MT1-MMP, and TIMPs 1-3. In situ hybridization was performed to localize MMP-2 and MMP-9 mRNA. The presence of active gelatinases was assessed using in situ zymography. RESULTS: Epithelial tumor cells were the main site of MMP-2, MMP-9, and MT1-MMP protein. Variable stromal cell localization was also observed, particularly in areas adjacent to tumor nests. Semiquantitative analysis revealed increases in MMP-9 and MMP-2 but not MT1-MMP staining scores in tumor epithelial cells in the transition from histologic Grade 1 to Grades 2 and 3. Matrix metalloproteinase-9 and MT1-MMP staining scores in tumor cells were significantly associated with the presence of myometrial invasion and vascular/lymphatic invasion, while MMP-2 did not correlate with these factors. In addition, MT1-MMP was co-localized with MMP-2, supporting its role in the activation of proMMP-2. Tumor cells from all histologic grades stained intensely for TIMP-2 and TIMP-3 proteins, while variable stromal staining was observed. In Grade 1 carcinomas TIMP-1 was predominantly immunolocalized to the stromal compartment with variable tumor cell localization being observed in Grades 2 and 3 carcinomas. Matrix metalloproteinase-9 and MMP-2 mRNAs were predominantly observed in tumor epithelial cells as well as in the stroma to varying degrees. In situ zymography revealed active forms of gelatinases at the cellular surface and in association with tumor epithelial cells within endometrial carcinoma tissues. CONCLUSIONS: These data suggest that increasing expression of MMPs and endometrial carcinoma progression are closely related. Active gelatinases are present in endometrial carcinoma, resulting in alterations to the microenvironment that promote tumor invasion and metastasis. PMID- 11920504 TI - Results of multimodality therapy for squamous cell carcinoma of maxillary sinus. AB - BACKGROUND: A wide variety of modalities, including surgery, radiation therapy, and chemotherapy, alone or in combination, have been used for the treatment of squamous cell carcinoma (SCC) of the maxillary sinus to obtain better local control and maintain functions. However, there is still much controversy with regard to the optimum treatment. METHODS: From 1987 to 1999, 33 patients with SCC of maxillary sinus were treated at the Department of Otolaryngology-Head and Neck Surgery, University of Tokyo Hospital. The treatment consisted of 30-40 grays (Gy) of preoperative radiotherapy with concomitant intraarterial infusion of 5 fluorouracil and cisplatin followed by surgery and 30-40 Gy of postoperative radiotherapy, for tumors without skull base invasion. For tumors invading the skull base, preoperative systemic chemotherapy with or without radiotherapy was performed, instead of intraarterial chemotherapy, then followed by skull base surgery. The surgical procedures varied according to the extent of tumor. Results were compared with those of the 108 patients treated in our hospital from 1976 to 1982. RESULTS: Partial maxillectomy was performed in 2 T2 patients and 12 T3 patients. Total maxillectomy was performed in 1 T2 patient, 3 T2 patients, and 7 T4 patients. Skull base surgery was performed in eight T4 patients. Orbital content and hard palate were preserved in 22 patients and 18 patients, respectively. The overall 5-year survival rates were 86% in T 3 patients and 67 % in T4 patients, respectively. CONCLUSIONS: Our multimodal treatment has provided favorable local control and survival outcome with good functional results. PMID- 11920505 TI - Circulating 92-kilodalton matrix metalloproteinase (MMP-9) activity is enhanced in the euglobulin plasma fraction of head and neck squamous cell carcinoma. AB - BACKGROUND: Cancer lethality is usually the result of local invasion and metastasis of neoplastic cells from the primary tumor. Because of their ability to degrade extracellular matrix components (EMC), matrix metalloproteinases (MMPs) have been implicated in the breakdown of basement membranes and underlying stroma, thereby facilitating tumor growth and invasion. METHODS: The authors quantitated, by gelatin zymography and densitometric analysis, MMP activity in the euglobulin plasma fraction of 50 healthy controls and 91 head and neck squamous cell carcinoma (HNSCC) patients (51 from the larynx and 40 from the oropharynx). RESULTS: The median value for 92-kilodalton (kD) MMP (MMP-9) activity was increased significantly in laryngeal (Md 2.1 arbitrary units (AU)/mL plasma; range, 0.2-6.4) and oropharyngeal patients (Md 2.08 AU/mL; range, 0.0 5.0) with respect to the controls (Md 0.48 AU/mL; range, 0.0-1.8). Both groups of cancer patients showed a similar behavior. Multivariate analysis indicated that circulating 92-kD MMP activity was not predicted by the clinical-pathologic parameters such as tumor stage, histologic grade, and metastatic lymph nodes. There was no association between high levels of MMP-9 activity and either cigarette smoking or alcohol consumption, major risk factors for developing HNSCC. CONCLUSIONS: The authors found a significant increase of MMP-9 plasma activity both in laryngeal and oropharyngeal squamous cell carcinoma patients as compared with healthy controls. Further studies are necessary to establish its usefulness in the clinical management of these patients. PMID- 11920506 TI - Hyperfractionated cyclophosphamide, vincristine, doxorubicin, and dexamethasone and highly active antiretroviral therapy for patients with acquired immunodeficiency syndrome-related Burkitt lymphoma/leukemia. AB - BACKGROUND: Patients with acquired immunodeficiency syndrome (AIDS)-associated lymphoma/leukemia have a poor prognosis and are frequently treated with low intensity therapy. The authors investigated the feasibility and efficacy of hyperfractionated cyclophosphamide, vincristine, doxorubicin, and dexamethasone (hyper-CVAD), a dose-intensive chemotherapy regimen, in patients with AIDS associated Burkitt lymphoma/leukemia, as well as the possible impact of highly active antiretroviral therapy (HAART) in these patients. METHODS: Thirteen patients with AIDS-associated Burkitt lymphoma (six patients) or leukemia (acute lymphoblastic leukemia; seven patients) were treated with hyper-CVAD alternating with high-dose methotrexate and ara-C for a total of eight cycles. Nine patients received HAART from the start of induction chemotherapy (seven patients) or later in the course of chemotherapy (two patients). The median patient age was 43 years (range, 32-55). Nine patients were diagnosed with human immunodeficiency virus (HIV) infection at the time of diagnosis of Burkitt lymphoma/leukemia; the other 4 patients had been diagnosed with HIV infection for a median of 37 months (range, 18-137) prior to the diagnosis of Burkitt lymphoma/leukemia. The median absolute CD4 count from the 9 patients with evaluable counts was 77 cells/microL (range, 9-544); only one patient had a count > 200/microL. RESULTS: Twelve patients (92%) achieved a complete remission (CR) and one achieved a partial response (PR). Eight patients continued in CR after a median of 31 months (range, 7-45) at the time of writing. Five patients were alive and in CR over two years later. The median survival was 12 months, with 48% of patients alive after 2 years. Six of seven patients who received HAART from the start of chemotherapy were alive and in CR after a median of 29 months (range, 7-45). The four patients who did not receive HAART died. The regimen was universally myelosuppressive, but the toxicity profiles, recoveries from myelosuppression, and incidences of infectious complications were similar to that of non-HIV patients with Burkitt lymphoma/leukemia treated with the same regimen. CONCLUSIONS: Hyper-CVAD is an effective regimen for patients with AIDS-associated Burkitt lymphoma/leukemia, with acceptable toxicity. The combination of hyper-CVAD and HAART is associated with long-term survival in patients with the two diseases, which, until recently, were both considered invariably fatal and almost futile to treat medically. PMID- 11920507 TI - Characteristics of indolent non-Hodgkin lymphoma in patients with type 1 human immunodeficiency virus infection. AB - BACKGROUND: There is recent evidence that the incidence of indolent non-Hodgkin lymphoma (NHL) appears to be increased in persons with the acquired immunodeficiency syndrome (AIDS). The current study was conducted to describe the clinical, immunologic, and pathologic characteristics of indolent B-cell lymphoma in patients infected with the human immunodeficiency virus (HIV). METHODS: The current report was a retrospective study of 10 cases of indolent NHL identified from the AIDS-Lymphoma Registry at the University of Southern California School of Medicine. These patients were compared with 336 consecutive patients with systemic intermediate/high-grade AIDS-related NHL who were diagnosed and treated at a single institution. RESULTS: The pathology of the indolent cases included follicular lymphoma (five patients), small lymphocytic lymphoma (two patients), and one case each of mucosa-associated lymphoid tissue (MALT), monocytoid B-cell, and marginal zone lymphoma. When comparing the indolent lymphomas with the intermediate/high-grade AIDS-NHL cases, no differences were observed with regard to demographic characteristics or history of prior opportunistic infection. HIV patients with indolent lymphomas were found to have a significantly higher median CD4+ lymphocyte count compared with patients with intermediate/high-grade NHL (531 /mm3 vs. 90 /mm3) (P < 0.0001). Bone marrow involvement was significantly more common in indolent NHL cases (50%) versus intermediate/high-grade NHL cases (17%) (P = 0.02). The median survival for patients with indolent NHL was significantly longer compared with patients with intermediate/high-grade NHL (66.8 months vs. 7.1 months) (P = 0.007). CONCLUSIONS: Indolent lymphomas occurring in patients infected with HIV appear to differ from intermediate/high grade lymphomas with regard to immune status and propensity for bone marrow involvement and prolonged survival. The median survival in the group of HIV seropositive patients with indolent NHL examined in the current study was found to be comparable to that reported in HIV-negative individuals. PMID- 11920508 TI - Helicobacter pylori infection in patients with gastric involvement by adult T cell leukemia/lymphoma. AB - BACKGROUND: Gastrointestinal involvement is seen frequently in patients with adult T-cell leukemia/lymphoma (ATLL). The authors previously showed a relatively low prevalence of Helicobacter pylori infection in individuals with human T-cell lymphotropic virus 1 (HTLV-1) infection, including patients with ATLL; however, the correlation between H. pylori infection and ATLL gastric involvement has not been investigated. METHODS: The authors studied 71 patients with ATLL. Gastric involvement was confirmed by endoscopy and biopsy. H. pylori infection was detected by serology, rapid urease test, and immunohistochemistry on biopsy samples. The expression of adhesion molecules on ATLL cells or their ligands on the vasculature in gastric mucosa was analyzed immunohistochemically. The expression of mucosal addressin cell adhesion molecule 1 (MAdCAM-1) was detected by reverse transcriptase-polymerase chain reaction (RT-PCR) analysis. RESULTS: Gastric involvement was detected in 21 patients (30%), including 8 patients with acute clinical subtype ATLL and 13 patients with lymphoma type ATLL. The prevalence of H. pylori infection was 86% (18 of 21 patients) in the patients with gastric involvement but only 38% (19 of 50 patients) in the patients without such involvement (P < 0.001). The expression of lymphocyte function-associated antigen 1 (LFA-1) and its ligand, intercellular adhesion molecule 1 (ICAM-1), was most frequent on ATLL cells infiltrating the stomach and was enhanced substantially on vascular endothelium in H. pylori-infected gastric mucosa. Human mucosal lymphocyte antigen 1 also was expressed on infiltrating ATLL cells in the stomach. The expression of MAdCAM-1 mRNA assessed by RT-PCR also was seen selectively in H. pylori-infected patients. CONCLUSIONS: ATLL cells infiltrate gastric tissues infected with H. pylori, probably through the interaction of adhesion molecules on these cells and their ligands on the vasculature, i.e., through the LFA-1/ICAM-1 pathway. PMID- 11920509 TI - Prognostic significance of Tie-1 protein expression in patients with early chronic phase chronic myeloid leukemia. AB - BACKGROUND: The Tie-1 tyrosine kinase receptor and its thus far unidentified ligand appear to play a distinct role in the regulatory pathways of early hematopoiesis and angiogenesis. Because vascularity is increased in the bone marrow of patients with chronic myeloid leukemia (CML), the authors evaluated the clinical significance of Tie-1 expression in such patients. METHODS: Using Western blot analysis and solid-phase radioimmunoassay (RIA), the authors quantified Tie-1 protein in bone marrow samples from 128 patients with CML and 31 normal controls. RESULTS: The median Tie-1 RIA value of CML samples was significantly higher than in normal controls (P = 0.01). The authors found no significant differences in Tie-1 levels in patients with early chronic, late chronic, accelerated, and blastic phases (P = 0.2). High Tie-1 levels correlated with short survival in patients with early chronic phase (P = 0.003; Cox proportional hazard model) but not in patients with late chronic (P = 0.2) or accelerated/blastic (P = 0.2) phase CML. Tie-1 protein level was also prognostic when patients were separated into two groups by the median value. High Tie-1 level in early chronic phase was associated with significantly shorter survival than low Tie-1 level (median survival, 116 vs. 61 months; P = 0.03). In patients with early chronic phase CML, Tie-1 levels correlated directly with patient age (P = 0.004) and platelet count (P = 0.003), and inversely with leukocyte count (P = 0.007). Tie-1 as a predictor of survival in early chronic phase CML was independent of risk group, spleen size, age, hemoglobin, and basophil count (P = 0.03; multivariate Cox proportional hazard model). CONCLUSIONS: The authors' findings support the hypothesis that angiogenesis may play a major role in the pathophysiology of chronic phase CML. PMID- 11920510 TI - Diagnostic utility of bilateral bone marrow examination: significance of morphologic and ancillary technique study in malignancy. AB - BACKGROUND: To retrospectively evaluate the significance of morphologic examination and ancillary studies performed on bilateral bone marrow biopsy specimens, 1864 bone marrow samples were studied. METHODS: Bilateral bone marrow biopsy specimens included 883 specimens that were evaluated for involvement by non-Hodgkin lymphoma (NHL); 381 specimens that were evaluated for involvement by carcinoma (CA); 362 specimens that were evaluated for involvement by Hodgkin disease (HD); 94 specimens that were evaluated for involvement by sarcoma (SA); 56 specimens that were evaluated for involvement by multiple myeloma (MM); 53 specimens that were evaluated for involvement by acute and chronic leukemia, myelodysplasia, and/or myeloproliferative disorders (LEUK); and 35 specimens that were evaluated for other reasons. RESULTS: Of all 1864 specimens, 410 samples (22.0%) were positive for disease, including 77% of MM samples, 58% of LEUK samples, 29.6% of NHL samples, 14% of SA samples, 9.9% of HD samples, and 6.8% of CA samples. A discrepancy between the left and right sides was identified in 48 specimens (11.7% of positive samples). The discrepancy rate was 39% for HD samples, 29% for SA samples, 23% for CA samples, and 9.2% for NHL samples. No morphologic discrepancies between bilateral samples were found in MM samples or LEUK samples. Bilateral flow cytometric studies (n = 113 samples) were positive in 11 samples (9.7%; all morphologically positive), with two discrepancies detected between bilateral samples. Bilateral cytogenetic studies (n = 74 samples) were positive in 5 samples (7%), and there were no discrepancies. Bilateral molecular studies (n = 16 samples) were positive in 7 samples (44%), and there were 3 discrepancies. CONCLUSIONS: Bilateral morphologic evaluation is useful in the evaluation of patients with NHL, HD, CA, and SA and is not indicated for patients with acute or chronic leukemia, myelodysplasia, MM, and other diseases. Bilateral flow cytometric or cytogenetic studies of bone marrow did not provide additional information in this population to justify bilateral samples. The role of bilateral molecular analysis needs to be defined further, but pooled samples for molecular studies may be adequate. PMID- 11920511 TI - Persistence of myeloma protein for more than one year after radiotherapy is an adverse prognostic factor in solitary plasmacytoma of bone. AB - BACKGROUND: Prognostic factors for solitary plasmacytoma of bone (SPB), whether measured before or after radiotherapy (RT), have not been established. The authors analyzed multiple factors for myeloma-free survival (MFS) and cause specific survival (CSS) in SPB patients treated with RT alone. METHODS: Between 1965 and 2000, 60 patients with carefully staged SPB were treated with RT alone at the M. D. Anderson Cancer Center. Patient ages ranged from 29-77 years (median, 54 years), and 75% of patients had a myeloma (M) protein in the blood and/or urine. No patients showed other lesions on skeletal survey or, in recent years, magnetic resonance imaging (MRI) of the spine; marrow aspirate was normal in all patients. Radiotherapy to the solitary lesion was given to a total dose of 30-70 Gy (median, 46 Gy). The authors analyzed the impact of multiple factors on MFS and CSS, including resolution v. persistence of M protein after RT, secretory v. nonsecretory disease at diagnosis, presence v. absence of an associated soft tissue mass on computed tomography or MRI scan, magnitude of serum M protein elevation at diagnosis, age, spinal v. nonspinal location, Karnofsky performance status, total RT dose, and tumor size. RESULTS: Median follow-up was 7.8 years (range, 1.0-25.5 years). On multivariate analysis, persistence of M protein more than one year after RT was the only independent adverse prognostic factor for MFS (P = 0.005) and CSS (P = 0.04). Most patients with M protein that persisted for more than one year after RT were diagnosed with multiple myeloma within 2.2 years of treatment. CONCLUSIONS: Patients with M protein that persists for more than one year after RT should be monitored frequently and considered for standard chemotherapy followed by intensive consolidation therapy when they either develop symptoms or show an increasing M protein level. PMID- 11920512 TI - Histopathology and MIB-1 labeling index predicted recurrence of meningiomas: a proposal of diagnostic criteria for patients with atypical meningioma. AB - BACKGROUND: Although various histopathologic features have been associated with aggressive behavior or recurrence of meningiomas, there is little agreement about which features are the most important and in what combination. The objective of this study was to formulate diagnostic criteria for atypical meningioma. METHODS: Eighty-three patients with meningiomas who underwent macroscopic total resection and had been followed until they developed recurrent disease or for at least 10 years were studied. Thirteen histopathologic parameters that related to recurrence of the tumor were evaluated in each patient. All possible combinations of histologic parameters that were related significantly to recurrence were used to formulate scoring models. The model that included the fewest parameters and that could distinguish tumor recurrence best within 10 years was chosen as the final model. RESULTS: The final model included three parameters: loss of architecture, mitoses > or= 1.5/mm2, and necrosis. Of the 52 tumors with a score < 2 (0 or 1 of the 3 parameters), all except 1 tumor did not recur within 10 years, and they were all considered benign meningiomas. Of the 31 tumors with a score > or = 2 (2 or 3 of the 3 parameters), 94% recurred within 10 years (76% recurred within 5 years), and they were considered atypical meningiomas. The estimated 5-year and 10-year recurrence rates for the benign meningiomas were 0.0% and 1.9%, respectively, for benign meningiomas and 71.0% and 93.5%, respectively, for atypical meningiomas (P < 0.001). The estimated 5-year and 10 year mortality rates also were significantly different (0.0% and 0.0% vs. 22.1% and 26.7%, respectively; P < 0.001). The MIB-1 labeling index (LI) for the entire group studied ranged from 0.4 to 33.5 (mean LI, 8.4). Fifty-two tumors with an LI of < 10 did not recur within 10 years. Of the 31 tumors with an LI > or = 10, 97% recurred (71% within 5 years). CONCLUSIONS: Histopathology and MIB-1 LI were able to predict clinical outcomes of patients with meningioma. The authors propose that atypical meningioma may be diagnosed when two of the following three criteria are present: loss of architecture, mitoses > or = 1.5/mm2, and necrosis. PMID- 11920513 TI - Primary central nervous system lymphomas: incidence and survival in the Southern and Eastern Netherlands. AB - BACKGROUND: An excessive increase in the incidence of primary central nervous system lymphoma (PCNSL) has been reported since the mid-1980s in the U.S. and U.K. Clinical studies have shown that radiotherapy and chemotherapy may prolong survival. In the current study, the authors describe the incidence, treatment, and survival of an unselected group of patients with PCNSL in the southern and eastern Netherlands. METHODS: Data regarding patients diagnosed between 1989-1994 were obtained from 4 population-based regional cancer registries in the southern and eastern Netherlands (n = 86) and the Eindhoven Cancer Registry for 1980-1988 (n = 6). Lymphomas were registered as PCNSL when a tissue diagnosis of CNS lymphoma was established for a patient with neurologic symptoms (i.e., lymphomas were not necessarily restricted to the CNS at the time of diagnosis). Only patients diagnosed during their lifetime with Stage I disease, Stage "IV" disease (i.e., diffuse CNS lymphoma), or disease of unknown stage were included (63 patients, 8 patients, and 15 patients, respectively, between 1989-1994). For 80 patients (93%) follow-up was complete until January 1, 1997. RESULTS: Between 1989-1994, an average World Standardized Rate of 2.3 cases and 1.7 cases per 1 million person-years, respectively, was reported for males and females. The median age of the patients at the time of diagnosis was 62 years, and was 66 years for patients with an unknown disease stage. In the area of the Eindhoven Cancer Registry the occurrence of PCNSL more than doubled from < 2% of all histologically confirmed primary CNS malignancies diagnosed between 1980-1985 to approximately 4% of cases diagnosed between 1986-1994. The median survival of all the patients was 4.1 months; the median survival was 5.8 months for patients with limited (Stage I and Stage IV) disease and was 0.6 months for patients with an unknown stage of disease. Approximately 65% of the patients with limited disease received radiotherapy and approximately 35% of such patients received chemotherapy. Furthermore, chemotherapy was given more often to patients age < 60 years who tended to have a slightly better survival than patients age > or = 60 years. CONCLUSIONS: The increase in the incidence of PCNSL in the 1980s may be explained in large part by changes in diagnostics and registration. The relatively high incidence and low survival rate of PCNSL in the southern and eastern Netherlands reported in the 1990s may be due in part to the inclusion of patients with systemic lymphoma and immunodeficiency disorders. However, a significant improvement in the prognosis of patients with PCNSL in the southern and eastern Netherlands diagnosed in the 1990s is unlikely. PMID- 11920514 TI - 31Phosphorus-magnetic resonance spectroscopy to assess histologic tumor response noninvasively after isolated limb perfusion for soft tissue tumors. AB - BACKGROUND: In patients with unresectable soft tissue sarcoma of the extremities, isolated limb perfusion (ILP) has been reported to result in significant tumor regression enabling limb-sparing resection in the majority of patients. However, clinical tumor response as evaluated by imaging and histopathology (extent of tumor necrosis) often differ significantly. The current study was initiated to evaluate prospectively the role of 31phosphorus-magnetic resonance spectroscopy (31P-MRS) in the noninvasive assessment of histologic response in patients treated with ILP. METHODS: Thirty-two patients with locally advanced and unresectable soft tissue tumors (sarcoma in 28 patients and bulky melanoma in 4 patients) were treated by ILP with recombinant human tumor necrosis factor-alpha and melphalan or with cytostatics. 31P-MRS was performed prior to treatment and at regular intervals after ILP until definite tumor resection. Clinical response parameters according to the World Health Organization as well as the histopathologic necrosis rate of the resection specimen were correlated with changes in the energy-rich phosphorous metabolites phosphocreatine (PCR); alpha-, beta-, gamma-adenosine triphosphate (ATP); phosphomonoesters (PME); and inorganic phosphate (Pi). RESULTS: Clinically, 15 of 32 patients (response rate [RR] of 47%) demonstrated a partial response (PR). The ratios of PME/PCR and PME/beta-ATP decreased significantly after ILP in comparison with preoperative values (P < 0.001). The changes in the PME/beta-ATP ratio were significantly different between clinical responders and nonresponders (P < 0.02) in contrast with the PME/PCR ratios (P < 0.09). Histologic necrosis of > 90% (pathologic (p) PR) was present in 17 resection specimens, 7 of which demonstrated no clinical response. Seven tumors demonstrated a pathologic complete response (pCR). When combining PR, pPR, and pCR (RR of 68%), 31P-MRS was able to predict response with a specificity of 94% and a sensitivity of 68% (P < 0.006, by the chi-square test). CONCLUSIONS: The considerable difference between clinical and pathologic RR after ILP underlines the shortcomings of established response criteria. Utilizing changes in PME/beta-ATP ratios, 31P-MRS is a highly specific tool with which to predict histologic response in this setting. This finding may be of major value in those patients in whom the decision to perform a major resection or amputation must be made for local tumor control. PMID- 11920515 TI - Immunohistochemical analysis and in situ hybridization of cyclooxygenase-2 expression in intraductal papillary-mucinous tumors of the pancreas. AB - BACKGROUND: The objective of this study was to investigate COX-2 expression in intraductal papillary-mucinous tumor of the pancreas (IPMT) using immunohistochemical staining (IH) and in situ hybridization (ISH). METHODS: Immunohistochemical staining of COX-2 was performed using samples from 42 patients with IPMT (hyperplasia, 10; adenoma, 13; noninvasive adenocarcinoma, 13; invasive adenocarcinoma, 6) and from 10 patients with ductal pancreatic adenocarcinoma, 10 with chronic pancreatitis, and 6 normal pancreatic tissues as controls. Also, COX-2 was determined in five patients with IPMT noninvasive adenocarcinoma, in whom all histologic types, hyperplasia, adenoma, and adenocarcinoma were observed in the same excised specimens. Furthermore, IH of proliferating cell nuclear antigen (PCNA) was performed, and the labeling index (LI) was calculated to investigate the correlation with COX-2. To confirm COX-2 mRNA, the authors performed ISH in 20 IPMT patients. RESULTS: COX-2 was positive in 0%, 0%, and 10% of pancreatic duct epithelial cells from normal pancreatic tissue, chronic pancreatitis, and IPMT hyperplasia, respectively. Whereas it was positive in 69%, 77%, 67%, and 80% of IPMT adenoma, IPMT noninvasive adenocarcinoma, IPMT invasive adenocarcinoma, and ductal pancreatic adenocarcinoma, respectively, showing significant differences between IPMT hyperplasia and IPMT adenoma or IPMT adenocarcinoma (noninvasive and invasive adenocarcinoma). In the same patient, COX-2 was negative in the hyperplasia region but positive in adenoma and adenocarcinoma regions, showing results reflecting the progression of the disease. In the COX-2 negative group, PCNA-LI was 19.2 +/- 17.9%, and 33.5 +/- 15.7% in the positive group, a significant difference. On ISH, COX-2 mRNA was expressed in three of four and seven of eight COX-2 positive patients with IPMT adenoma and adenocarcinoma, respectively. CONCLUSIONS: COX-2 was highly expressed in adenoma and adenocarcinoma in IPMT, showing a relation to the histologic grade of IPMT. PMID- 11920516 TI - Morphologic features of neuroblastoma (Schwannian stroma-poor tumors) in clinically favorable and unfavorable groups. AB - BACKGROUND: After the establishment of the International Neuroblastoma Pathology Classification system, the authors studied retrospectively the prognostic impact of morphologic features in a series of two clinically distinct subsets of patients with peripheral neuroblastic tumors (NTs), i.e., tumors in the neuroblastoma category. METHODS: Forty-seven NTs categorized into either clinically favorable or unfavorable subgroups were selected randomly from 100 NTs for a histologic review that included the evaluation of 14 morphologic characteristics. The review was performed individually followed by a group review. The correlations of the prognostic significance of the individual morphologic features and the correlations among them were determined by use of odds ratios (ORs) with corresponding 95% confidence intervals (95%CIs). The inter rater agreement was determined by using the Cohen kappa coefficient. RESULTS: Ten of 14 morphologic features, including nuclear size, cellularity, prominent nucleoli in undifferentiated or poorly differentiated neuroblasts, and the number of mitotic and karyorrhectic cells (MKI), showed a significant correlation with the clinical groups (ORs between 36.9 and 10.5 and P values between < 0.001 and 0.002). In addition to the patient's age at diagnosis (OR, 7.4; 95%CI, 1.9-28.9; P = 0.002), 8 of 14 features also provided prognostic information (ORs between 35.1 and 7.9 and P values between < 0.001 and 0.039). CONCLUSIONS: This study again confirmed the prognostic impact of the criteria used in the Shimada system and revealed that some other morphologic features, such as prominent nucleoli in undifferentiated and poorly differentiated neuroblasts, identify unfavorable tumor biology, partly independent from the patient's age at diagnosis. However, the prognostic impact of these features needs to be confirmed by analysis of a large series of neuroblastic tumors. PMID- 11920517 TI - Intra-tumoral interleukin-6 down-regulation system and genetic mutations of tumor suppressor genes in colorectal carcinoma. AB - BACKGROUND: The interleukin (IL)-1-IL-6 network, the most potent cascade of pro inflammatory cytokines, plays an autocrine role in tumor growth. The IL-1-IL-6 network is down-regulated by a phased cytokine inhibitor IL-1 receptor antagonist (ra) and an anti-inflammatory cytokine IL-10. The current study evaluated this down-regulation system in colorectal carcinoma and its relation to the genetic alteration of tumor suppressor genes. METHODS: Seventy-four specimens of primary colorectal carcinoma and normal mucosa were collected to measure tissue concentrations of cytokines. Polymerase chain reaction amplification was performed to investigate the loss of heterozygosity of the microsatellite markers on chromosomes 17p and 18q. RESULTS: The IL-1ra/IL-6 ratio in the carcinoma specimens was lower than ratios in adenomas and normal mucosae and decreased with disease progression. The IL-1ra/IL-6 ratio in early cancers tended to be lower than that in adenomas and normal mucosae. However, the tissue concentrations of IL-1beta and IL-10 were not associated with any clinicopathologic parameters. The tissue IL-1ra concentration correlated with that of IL-6 only in adenomas and early cancers. Immunohistochemically, IL-1ra and IL-6 were localized in the tumor cytoplasm. A reduced tissue IL-1ra/IL-6 ratio in the carcinomas correlated with poor prognosis and was associated with the loss of heterozygosity of the microsatellite markers on chromosomes 18q. CONCLUSIONS: There is an IL-6-IL-1ra network system in colorectal tumors, but this system deteriorates with carcinogenesis and tumor growth. The deterioration of this network system was associated with the allelic loss of a portion of chromosome 18q, reflecting the genetic alteration of tumor suppressor genes. PMID- 11920518 TI - Monoclonal antibodies to target epidermal growth factor receptor-positive tumors: a new paradigm for cancer therapy. AB - BACKGROUND: Traditional cytotoxic approaches to tumor management are associated with efficacy and toxicity limitations. Blockade of the epidermal growth factor receptor (EGFR) and its ligands is a novel approach to the treatment of human tumors that offers a noncytotoxic alternative to cancer treatment. METHODS: An English-language literature search was conducted to identify studies assessing the in vitro and in vivo effects of EGFR blockade with an emphasis on approaches that use monoclonal antibody therapy. RESULTS: The EGF pathway regulates normal cellular processes and appears to be correlated with the development of malignancy. Approximately 30% of human tumors express EGFR, which has been reported to be correlated with poor prognosis and diminished disease-free and overall survival in selected tumor types. A number of anti-EGFR monoclonal antibodies have been developed, which currently are undergoing clinical trials in humans. Effective anti-EGFR monoclonal antibodies compete with endogenous ligands, primarily EGF and transforming growth factor-alpha, for receptor ligand binding sites. Binding to EGFR blocks critical signaling pathways and interferes with the growth of tumors expressing EGFR. Anti-EGFR monoclonal antibodies that currently are under study include IMC-C225, EMD 55900, ICR 62, and ABX-EGF. CONCLUSIONS: These antibodies have demonstrated promising results and appear to have been well tolerated. EGFR-targeted therapy addresses important, unmet needs in the treatment of human tumors, particularly EGFR-positive epithelial tumors including common malignancies of the head and neck, lung, and colon. PMID- 11920519 TI - Postmenopausal cancer risk after self-reported endometriosis diagnosis in the Iowa Women's Health Study. AB - BACKGROUND: Endometriosis, the presence of endometrial tissue outside the uterus, has an estimated prevalence between 4% and 10%. A recent study reported that women with a hospital discharge diagnosis of endometriosis were at increased risk of cancer at any site, breast cancer, ovarian cancer, and hematopoietic malignancies, especially non-Hodgkin lymphoma (NHL). METHODS: The authors examined whether self-reported diagnosis of endometriosis was associated with increased risk of various cancers in the Iowa Women's Health Study. Incident cancer cases were identified from 1986 through 1998. Cox proportional hazards regression models were used to calculate relative risks and 95% confidence intervals for the particular cancers of interest. RESULTS: Of 37,434 participants in this analysis, 3.8% reported a history of endometriosis at baseline in 1986. After 13 years of follow-up, 1795 breast, 188 ovarian, and 243 NHL cases were detected in the cohort. Endometriosis was not associated with risk of all cancers combined (age-adjusted relative risk [RR], 0.9; 95% confidence interval [CI], 0.7 1.2), breast carcinoma (RR, 1.0; 95% CI, 0.8-1.3), or ovarian carcinoma (RR, 0.8; 95% CI, 0.2-2.4). However, endometriosis was significantly associated with risk of NHL (age-adjusted RR, 1.8; 95% CI, 1.0-3.0), especially diffuse NHL (RR, 3.2; 95% CI, 1.8-5.6). Multivariate adjustment had minimal effect on the point estimates of risk (NHL RR, 1.7; 95% CI, 0.97-2.7; diffuse NHL RR, 3.3; 95% CI, 1.9-5.9). Endometriosis was not associated with elevated risk of lung, urinary tract, endometrial, melanoma, or colorectal carcinomas (RRs, 1.2, 0.8, 1.2, 0.7, and 0.7, respectively). CONCLUSIONS: These results corroborate a previously reported association between endometriosis and increased risk of NHL but not cancer at other sites. PMID- 11920520 TI - Incidence of the superficial fascia and its relevance in skin-sparing mastectomy. AB - BACKGROUND: With the move away from classical radical mastectomy to ever more skin-sparing procedures, there has been an ongoing discussion about how much skin and subcutaneous tissue should be resected to perform an adequate mastectomy while leaving viable skin flaps. One of the common recommendations is to dissect just superficial to the superficial layer (SL) of the superficial fascia of the breast. This, in turn, has revived the old, unsolved controversy about the existence or absence of the SL, a fascia that reportedly encloses the mammary gland ventrally. In skin-sparing mastectomies (SSM), which combine tumor resection with immediate breast reconstruction, the ideal would be to create skin flaps that are thin enough to remove all breast tissue but at the same time are thick enough to preserve flap circulation. The feasibility of meeting these two goals simultaneously and the possible role and relevance of the SL as a guide to dissection in SSM was examined in this study. METHODS: Sixty-two breast resection specimens from 31 women who underwent breast reduction were examined histologically to determine whether the SL was present, whether breast tissue could be detected within or beyond this SL, the measured distance between the caudal border of the dermis and the SL or the breast tissue, and whether the thickness of the subcutaneous fat layer was correlated with the patients' physical data, such as body weight or body mass index (BMI). RESULTS: The SL was absent in 44% of resection specimens. When the SL was present, 42% of specimens contained several islands of breast tissue within the SL. No breast tissue was found beyond the SL. The minimal distance between the SL and the dermis varied from 0.2 mm to 4.0 mm; the minimal distance between the breast tissue and the dermis was 0.4 mm. In 50% of specimens, the minimal distance between the dermis and the SL or breast tissue was < 1.1 mm. A distance of > or = 5 mm was encountered in only 17% of specimens, and a distance of > or = 10 mm was encountered in only 5% of specimens. No significant correlation between the right and left breast was found with any of the parameters examined. A weak negative correlation was seen between the BMI and the mean thickness of the subcutaneous fat (P = 0.049; correlation coefficient [r] = -0.39; Spearman rank correlation). CONCLUSIONS: Histologic evaluation revealed that the SL is not present in all breasts and, thus, cannot serve as a reliable plane of dissection. Furthermore, if the SL is present microscopically, then it often is too thin and delicate to be detectable macroscopically. Finally, even if the SL is present and visible macroscopically, the distance to the overlying skin is so small in the majority of patients that a dissection superficial to the SL would not leave viable skin flaps in skin-sparing mastectomies. PMID- 11920521 TI - The association between breast carcinoma and meningioma in women. AB - BACKGROUND: Published case reports of a possible association between meningioma and breast carcinoma are not uncommon in the literature. Four published analytic studies have addressed this suggested association specifically. Three of these studies reported significant associations, with relative risk estimates mostly between 1.5 and 2.0. The other study reported relative risk point estimates near 1.5, but confidence intervals included 1.0. The current study was a population based, retrospective cohort analysis that evaluated the risk of subsequent breast carcinoma in women who were diagnosed with meningioma and the risk of subsequent meningioma in women who were diagnosed with breast carcinoma. METHODS: The measure of association is the relative risk and is reported as the standardized incidence ratio (SIR). Using western Washington State cancer registry data and intercensal population estimates for western Washington State, incidence rates of second primary tumor were compared between identified meningioma and breast carcinoma cohorts and the general population for the years 1992-1998. RESULTS: The risk of breast carcinoma after patients were diagnosed with meningioma (SIR) was 1.54 (95% confidence interval [95% CI], 0.77-2.75). The risk of meningioma after patients were diagnosed with breast carcinoma was 1.40 (95% CI, 0.67-2.58), and the risk of meningioma after patients were diagnosed with invasive breast carcinoma was 1.64 (95% CI, 0.79-3.02). In each combination for age groups ages > 50 years, risks were elevated, but the confidence intervals included 1.0. CONCLUSIONS: These results suggest that the risk of meningioma among women who have experienced breast carcinoma and the risk of breast carcinoma among women who have experienced meningioma are elevated moderately. Shared risk factors may account for the relatively week bidirectional associations seen in this and other studies. PMID- 11920522 TI - Expression of tumor rejection antigens in colorectal carcinomas. AB - BACKGROUND: The authors recently reported that the SART2 and SART3 antigens encode tumor epitopes recognized by HLA-A24-restricted and tumor-specific cytotoxic T lymphocytes (CTLs) established from esophageal carcinoma patients. The current study investigated these antigens to explore a potential molecule for specific immunotherapy for colorectal carcinoma patients. METHODS: The SART2 and SART3 antigens were investigated by Western blotting in colorectal carcinoma cell lines and in cancer tissues. For induction of CTLs, peripheral blood mononuclear cells (PBMCs) of HLA A-24-positive cancer patients were stimulated in vitro with peptides. RESULTS: The 140 kD SART3 antigen was expressed in both the cytosol and nuclear fractions of all six colon carcinoma cell lines, 27 of 41 (65.9%) cytosol fractions, 30 of 41 (73.2%) nuclear fractions of colorectal carcinoma tissue samples, and in 0 of 7 non-tumorous tissues. The 100 kD SART2 antigen was expressed in the cytosol fractions of 2 of 6 colon carcinoma cell lines, 5 of 20 (25%) cytosol fractions of colorectal carcinoma tissue samples, and in 0 of 7 non tumorous tissues. HLA-A24-restricted CTLs cytotoxic to colon carcinoma cells were induced from PBMCs of colon carcinoma patients by stimulation with the two immunogenic peptides of SART3. CONCLUSIONS: The SART3 antigen could be an appropriate target molecule for specific immunotherapy for colorectal carcinoma patients. PMID- 11920523 TI - Immunohistochemically detected hepatic micrometastases predict a high risk of intrahepatic recurrence after resection of colorectal carcinoma liver metastases. AB - BACKGROUND: Hepatic metastases from colorectal carcinoma frequently recur after resection and hepatic micrometastases most likely are important in the development of such recurrences. The objectives of the current study were to assess the feasibility of the immunohistochemical detection of hepatic micrometastases from colorectal carcinoma and to determine their clinical significance. METHODS: Fifty-three patients underwent curative hepatic resection for colorectal carcinoma metastases. Multiple tissue sections were cut from the advancing margin of the largest hepatic metastasis in each patient and were stained with an antibody against cytokeratin-20 to detect hepatic micrometastases, which were defined as discrete microscopic cancerous lesions surrounding the dominant metastasis. RESULTS: Normal hepatocytes and intrahepatic bile duct epithelia stained negative for cytokeratin-20 in all patients, whereas the largest hepatic tumors stained positive in 46 patients (86.8%). Among the 46 patients with hepatic tumors that were positive for cytokeratin-20, hepatic micrometastases were found immunohistochemically in 32 patients (69.6%). The presence of hepatic micrometastases was associated with a larger number of macroscopic hepatic metastases (P = 0.047) and patients with hepatic micrometastases were found to demonstrate a higher probability of intrahepatic recurrence (P = 0.003) compared with those patients without hepatic micrometastases. In addition, patients with hepatic micrometastases demonstrated a worse survival (10-year survival rate of 21.9%) compared with those patients without hepatic micrometastases (10-year survival rate of 64.3%) (P = 0.017). CONCLUSIONS: Immunohistochemical detection of hepatic micrometastases is feasible in patients with colorectal carcinoma liver metastases. Hepatic micrometastasis indicates widespread hepatic involvement and thus predicts an increased risk of intrahepatic recurrence after hepatic resection and a poorer patient prognosis. PMID- 11920524 TI - Seminal vesicle involvement in patients with D1 disease predicts early prostate specific antigen recurrence and metastasis after radical prostatectomy and early androgen ablation. AB - BACKGROUND: Controversy persists regarding the management of patients who present with locally advanced metastatic prostate carcinoma. Although radical prostatectomy is not curative, there is growing evidence that survival may be prolonged when the surgery is combined with early androgen ablation. In the current study, the authors present data with which to evaluate and define factors for disease progression in patients undergoing radical prostatectomy with lymph node positive disease who are treated with early endocrine ablation. METHODS: Data from 40 patients undergoing radical prostatectomy and early androgen ablation between 1987-1998, all of whom had lymph node positive disease, were analyzed. Age, preoperative prostate specific antigen (PSA) level, clinical and pathologic Gleason score, surgical margin, seminal vesicle involvement (SVI), and the number and percentage of involved positive lymph nodes were analyzed to predict PSA progression, metastasis, and death using univariate and multivariate statistical techniques. RESULTS: Univariate analysis identified only SVI as a statistically significant predictor of PSA progression and metastasis. Twenty seven patients (67.5%) were found to have SVI. Multivariate analysis failed to identify other factors that added significantly to the predictive ability of SVI. Kaplan-Meier estimates of time to PSA recurrence and metastasis demonstrated that SVI was highly predictive of disease progression. The median time to PSA progression for the 27 patients with SVI was 7.5 years compared with no progression reported in the 13 patients without SVI (P = 0.011). CONCLUSIONS: VI is a very powerful predictor of disease progression in patients with lymph node positive disease who undergo radical prostatectomy and early androgen ablation. In the current study, preoperative PSA, clinical or pathologic Gleason scores, and other clinical factors were not found to be predictive of disease outcome. PMID- 11920525 TI - A multiinstitutional, concurrent chemoradiation trial of strontium-89, estramustine, and vinblastine for hormone refractory prostate carcinoma involving bone. AB - BACKGROUND: Estramustine phosphate (EMP) and vinblastine have radiosensitizing properties and significant activity against hormone refractory prostate carcinoma. Strontium-89 is a palliative agent that acts as a selective radiation source for bone metastasis. The combination of EMP, vinblastine, and strontium-89 was developed to exploit the potential for radiosynergy. PATIENTS AND METHODS Forty-four patients at the Brown Oncology Group affiliated hospitals were treated with oral EMP 600 mg/m2 daily on Weeks 1-4 and 7-10, vinblastine 4 mg/m2 intravenously once each week on Weeks 1-4 and 7-10, and strontium-89 2.2 MBq/kg on Day 1. Courses were repeated every 12 weeks. Response assessment was based on a change in the serum prostate specific antigen (PSA) levels, correlated with change in measurable disease and bone scan appearance. RESULTS: A greater than or equal to 50% decline in PSA for at least 6 weeks was observed in 21 patients (48%, 95% confidence interval, 33-62%). Median duration of response was 23 weeks (range, 6-70.8 weeks). The median survival was 13 months with 1- and 2-year survival rates of 55% and 25%, respectively. After completion of protocol therapy, a retrospective review showed that only nine patients received subsequent palliative external beam radiation after progression. CONCLUSIONS: The addition of strontium-89 to the regimen of EMP and vinblastine can be delivered safely and in repeated doses, provides effective palliation, and may decrease the need for future radiation therapy. A randomized trial is necessary to quantify these effects. PMID- 11920526 TI - Prospective study of cancer detection in black and white men with normal digital rectal examination but prostate specific antigen equal or greater than 4.0 ng/mL. AB - BACKGROUND: The serum prostate specific antigen (PSA) concentration with no clinical evidence of prostate carcinoma is higher and more variable in black than in white American men. The influence of this phenomenon on relations between race, PSA, and cancer detection in men with a PSA greater than or equal to 4.0 ng/mL has not been investigated. METHODS: Between January 1992 and December 2000, 451 black and 480 white men with a normal digital rectal examination and a PSA greater than or equal to 4.0 ng/mL had an initial prostate biopsy at one medical center. The histology of the biopsy specimens and the Gleason score of malignant specimens was determined by one uropathologist. RESULTS: Cancer was detected in 207 (46%) black and 167 (35%) white men (P = 0.0006). When adjusted for PSA, cancer detection was also greater in the black than the white men, but the difference did not achieve statistical significance (relative risk, 1.30; 95% confidence interval [CI], 0.99-1.71; P = 0.06). Gleason score 7-10 cancer was detected in 88 (20%) black and 45 (9%) white men (P = 0.0001), and the difference remained significant when adjusted for PSA (relative risk, 1.73; 95% CI, 1.16 2.61; P = 0.0008). In the intermediate PSA range of 4.0-9.9 ng/mL, cancer detection and Gleason score 7-10 cancer detection was greater in black than in white men younger than 60, 60-69, and 70 years of age or older, but the difference was significant only for Gleason score 7-10 cancer detection among men 60-69 years of age (P = 0.006). CONCLUSIONS: There is a direct correlation between Gleason score and cause specific survival with local stage prostate carcinoma. The authors' study indicates that prostate carcinomas with established malignant potential are more likely to be identified in black than in white men with PSA elevation as the only indication of malignancy and raises the possibility that a PSA threshold less than 4.0 ng/mL in black men younger than 70 years of age may reduce racial disparities in prostate carcinoma morbidity and mortality. PMID- 11920527 TI - Metastatic nonseminomatous germ cell tumors of the testis: results of elective and salvage surgery for patients with residual retroperitoneal masses. AB - BACKGROUND: A mass may persist in the para-aortic region after patients undergo chemotherapy for metastatic, nonseminomatous germ cell tumor of the testis (NSGCT). Retroperitoneal lymphadenectomy removes the mass, which may contain residual active malignancy, and allows histologic assessment of the effectiveness of the chemotherapy. Whereas some have favored early, elective removal of such masses, others have chosen to observe them, reserving salvage surgery for patients who experience disease recurrence. A retrospective analysis was undertaken to define the outcome in these two groups of patients. METHODS: After receiving chemotherapy for metastatic NSGCT, 442 men underwent lymphadenectomy for residual masses (measuring > or = 1 cm in greatest dimension) between 1976 and 1999, inclusive. Three hundred thirty men underwent elective surgery within 3 months of the completion of chemotherapy, and 112 men underwent salvage surgery after receiving reinduction chemotherapy for tumor recurrence. RESULTS: The residual mass was removed completely in 87% and 72% of patients in the elective and salvage lymphadenectomy groups, respectively; was removed with difficulty and possibly incompletely in 9% and 21% of patients, respectively; and was definitely removed incompletely in 4% and 7% of patients, respectively. The operative mortality rate was 0.9% in the elective surgery group and 1.8% in the salvage surgery group. There was malignant teratoma, undifferentiated in 8.5% of patients in the elective surgery group and in 49% of patients in the salvage surgery group (P < 0.001). Differentiated teratoma and necrosis/fibrosis were present in 66.0% and 25.4% of patients in the elective surgery group, respectively, and in 38.4% and 12.5% of patients in the salvage surgery group, respectively. The authors were unable to produce a clinically useful model to predict the presence of necrosis/fibrosis only in either group. The 5-year recurrence free and overall survival rates were 83% and 89%, respectively, in the elective surgery group and 62% and 56%, respectively, in the salvage surgery group. For the salvage surgery group, the completeness of surgical excision and the presence of undifferentiated teratoma were of overriding importance for overall survival. A variety of other patient-related, tumor-related, and surgery-related factors also were significant in the final model for the elective surgery group. CONCLUSIONS: The current results demonstrate the low level of morbidity that can be obtained, even in the salvage surgery group, and the importance of complete surgical resection in this setting. Because it is not possible to predict with sufficient accuracy which patients will have favorable pathology (necrosis/fibrosis), the authors continue to recommend elective surgery for all suitable men with residual masses after they receive first-line chemotherapy. PMID- 11920528 TI - Multivariate analysis of survival, recurrence, progression and development of mestastasis in T1 and T2a transitional cell bladder carcinoma. AB - BACKGROUND: Determination of prognosis factors associated with survival, recurrence, progression, and development of metastasis in T1 and T2a transitional cell carcinoma (TCC) of the bladder is discussed. METHODS: A study was conducted of a group of 210 patients with primary bladder TCC at classification T1 (n = 175) and T2aN0M0 (n = 35). A total of 177 variables were studied in each patient. The monoclonal antibodies used were the following: DO7 (p53) and MIB-1 (Ki-67). Prognosis was obtained using Kaplan-Meier methodology and Cox proportional hazards model. RESULTS: The average follow-up period was 6.7 years. Cancer related survival rates at 5 and 10 years were 82.96% and 74.78%, respectively. The independent survival variables were the following: age and expression of p53. Recurrence free survival at 5 and 10 years stood at 51.80% and 42.71%, respectively. The independent recurrence variables were T2a classification, tumor multifocality, tumor size of greater than 3 cm, carcinoma in situ in random biopsy, and expression of Ki-67. Progression free survival rates at 5 and 10 years were 75.31% and 69.16%, respectively. The independent progression variables were age, T2a classification, and expression of p53. Metastasis free survival rates at 5 and 10 years stood at 87.23% and 84.55%, respectively. The expression of p53 was the sole variable to provide an independent prediction of metastasis. CONCLUSIONS: The expression of p53 clearly has an independent effect on the prediction of survival, progression and development of metastasis, showing a dose response effect. Tumor multifocality and T2a classification are the variables that best predict recurrence. PMID- 11920529 TI - Prostate specific antigen complexed to alpha-1-antichymotrypsin in patients with intermediate prostate specific antigen levels. AB - BACKGROUND: The authors attempted to evaluate prospectively the usefulness of serum prostate specific antigen (PSA) complexed to alpha-1-antichymotrypsin (PSA ACT) in the early detection of prostate carcinoma and its ability to discriminate between prostate carcinoma and benign prostatic hyperplasia (BPH), especially among patients with intermediate PSA levels. METHODS: Between December 1999 and August 2000, systematic sextant biopsies were performed on 281 prospective patients with prostate carcinoma who had serum PSA levels between 4.1 ng/mL and 20.0 ng/mL. The serum samples were assayed by using kits that were designed specifically for measuring serum PSA, PSA-ACT, and free PSA levels. The clinical values of PSA, PSA-ACT, the free PSA to total PSA ratio (F/T ratio), the free PSA to PSA-ACT ratio, PSA density (PSAD), and PSA-ACT density (ACTD) were compared by using receiver operating characteristic (ROC) curve analysis. RESULTS: Biopsy yielded no evidence of malignancy in 198 patients, and prostate carcinoma was confirmed in 83 patients. ROC analysis demonstrated that the area under the curve (AUC) for PSA-ACT was greater than that for total PSA and was equivalent to that for the F/T ratio in both groups of patients (PSA ranges of 4.1-20.0 ng/mL and 4.1-10.0 ng/mL, respectively). The AUC for the ACTD was greater than the AUC for the PSAD and had the highest value of all parameters. CONCLUSIONS: The measurement of PSA-ACT represents an alternative to the use of total and free PSA. The ACTD value is the most useful for discriminating between BPH and prostate carcinoma. PMID- 11920530 TI - The dynamics of prostate specific antigen during watchful waiting of prostate carcinoma: a study of 94 Japanese men. AB - BACKGROUND: For the moment, there is uncertainty about the usefulness of early treatment of localized prostate carcinoma, uncertainty about whether some patients with early cancer can be managed expectantly, and uncertainty about how such patients might be recognized. METHODS: The authors studied serial values of prostate specific antigen (PSA) in 94 Japanese men with diagnosed prostate carcinoma and who were managed by watchful waiting. Their median follow-up duration was 32 months (range, 1.6-118). The authors used a log-linear model to fit the values of PSA over time, and then they used the Cox survival model to relate the intercept (PSA amplitude) and slope (relative velocity) to observed local or systemic outcomes that were independent of PSA. RESULTS: The authors found that the log-linear model fit the serial values of PSA during watchful waiting very well. Prostate specific antigen amplitude related significantly to T classification (P = 0.0006), but not to grade (P > 0.2), and the relative velocity related significantly to both T classification (P = 0.009) and to grade (P = 0.02). Although the T classification, histologic grade, and log(PSA) at diagnosis were associated significantly with time to outcome, the combination of amplitude and relative velocity provided more information. These 2 PSA parameters resulted in a higher model likelihood ratio, and their individual P values in the Cox model were 0.0005 and 0.005, respectively. With these two in the Cox model, T classification, grade, log(PSA), and PSA doubling time provided no further significant information. CONCLUSIONS: A log-linear model seems to fit serial measurements of PSA during watchful waiting, and preliminary results suggest that both the amplitude and the relative velocity relate closely to clinical outcomes. PMID- 11920531 TI - Direct visual inspection for cervical cancer screening: an analysis of factors influencing test performance. AB - BACKGROUND: The authors evaluated direct visual inspection of the cervix after the application of 5% acetic acid (DVI) as a cervical cancer screening test for use in low-resource settings. METHODS: Four tests were used to screen 2754 previously unscreened women: DVI with and without x4.5 magnification and differentiating between all lesions and well-circumscribed lesions; testing for high-risk types of human papillomavirus DNA using the Hybrid Capture II assay, cervical cytology, and Cervicography. Women with positive results on any of the four screening tests were referred for colposcopy and histologic sampling. All women were tested for N. gonorrhea, C. trachomatis, T. vaginalis, and human immunodeficiency virus 1. RESULTS: Histologically confirmed carcinoma was diagnosed in 21 women (0.8%), high-grade squamous intraepithelial lesions (SILs) were diagnosed in 96 women (3.5%), and low-grade SILs were diagnosed in 102 women (3.7%). The estimated sensitivity of DVI when performed without magnification for high-grade SILs was 70%, with an estimated specificity of 79%. Magnification did not significantly improve sensitivity for high-grade SILs (74% with magnification) but significantly reduced specificity (77%). Restricting the definition of a positive DVI test to a well-defined acetowhite lesion reduced sensitivity and significantly improved specificity. Infection with T. vaginalis, N. gonorrhea, and C. trachomatis did not alter sensitivity or specificity. CONCLUSIONS: This study confirmed the utility of DVI as a primary screening test. Evaluation of the clinical effectiveness of screening with this low-cost and simple test in low-resource settings is necessary. PMID- 11920532 TI - The outcome of endometrial carcinoma surveillance by ultrasound scan in women at risk of hereditary nonpolyposis colorectal carcinoma and familial colorectal carcinoma. AB - BACKGROUND: Endometrial carcinoma is the most common extracolonic malignancy associated with hereditary nonpolyposis colorectal carcinoma syndrome (HNPCC). The risk of endometrial carcinoma in HNPCC mutation carriers is approximately ten times that of the general population, and endometrial ultrasound surveillance to detect early cancer in asymptomatic individuals is recommended by the International Collaborative Group on HNPCC. There is little, if any, published data addressing the effectiveness of surveillance in HNPCC and familial colorectal carcinoma. METHODS: The outcomes of endometrial carcinoma surveillance scans were collected from the St Mark's Hospital Imperial Cancer Research Fund Family Cancer Clinic in the UK and the Netherlands Foundation for the Detection of Hereditary Tumors. Two hundred ninety two women from HNPCC (171) or HNPCC-like (98) families between the ages of 25-65 years were offered pelvic ultrasound surveillance scans for a period of up to 13 years. RESULTS: Results were available from 269 women. The study period included a total of 825.7 years of risk. Two cases of endometrial carcinoma were reported. Neither case was detected by surveillance scanning. Both cases presented at an early stage with symptoms and were subsequently cured. CONCLUSIONS: Endometrial carcinoma surveillance in hereditary colorectal carcinoma may not offer obvious clinical benefits. PMID- 11920533 TI - Transcervical needle biopsy for the differential diagnosis between uterine sarcoma and leiomyoma. AB - BACKGROUND: The clinical differential diagnosis between uterine sarcoma and benign leiomyoma is difficult even with magnetic resonance imaging (MRI). Therefore, a considerable number of patients have undergone hysterectomies due to an indication of "suspected malignancy" based on tumor size alone. However, approximately 80% of these hysterectomies have been judged to have been recommended inappropriately. In such situations, reliable preoperative diagnostic tests are required. The authors have evaluated the accuracy of needle biopsy for uterine myoma-like tumors, a procedure that to the authors' knowledge has been performed infrequently. METHODS: Transcervical needle biopsy was performed in 435 patients with uterine myoma-like tumors. The biopsy specimens were scored for degree of malignancy according to the histopathologic criteria proposed by Bell et al. Histopathologic evaluation of surgical specimens and clinical outcome after 2 years of follow-up were used as the reference standards. RESULTS: Of 435 patients, 7 had uterine sarcomas, 4 of which were scored as > or = 4 points and were diagnosed as "sarcoma" by needle biopsy alone. No sarcoma cases were included in the group of patients with a score of 0. The cutoff score combining the highest sensitivity and specificity with respect to distinguishing uterine leiomyosarcoma from uterine leiomyoma was 2; sensitivity, specificity, and positive and negative predictive values were 100%, 98.6%, 58%, and 100.0%, respectively. CONCLUSIONS: Transcervical needle biopsy using histopathologic scoring is a reliable diagnostic test for the differential diagnosis between uterine sarcoma and leiomyoma. This diagnostic method, combined with MRI screening, could reduce the number of patients currently undergoing unnecessary surgery. PMID- 11920534 TI - The clinical and diagnostic relevance of CD23 expression in the chronic lymphoproliferative disease. AB - BACKGROUND: CD23 antigen is a cell surface protein considered important in the differentiation of chronic lymphocytic leukemia (CLL) from other lymphoid leukemias. METHODS: To better clarify CD23 role as a diagnostic tool, the authors retrospectively evaluated clinical and laboratory features of 372 patients who were referred to M.D. Anderson Cancer Center with a diagnosis of CLL or B-cell chronic lymphoproliferative disease. RESULTS: Most of the patients (91%) were CD19+/CD5+. Only 6% of these CD19+/CD5+ patients were CD23-. Overall, CD23- patients had the worse prognostic features compared with CD23+ cases, including anemia (P = 0.03), massive splenomegaly (P = 0.000), high lactate dehydrogenase (P = 0.007), high beta2-microglobulin (P = 0.006), older age (P = 0.001), and male gender (P = 0.02). Surface immunoglobulin expression was moderate/strong in 19 (82%) patients, and FMC-7 was positive in 22 (96%) patients. None of the 13 patients tested for CD10 expressed the antigen. Based on morphology, of the CD23, 16 (70%) were diagnosed with mantle cell leukemia (MCL) was diagnosed in 16 (70%) CD23- patients, 3 (13%) with splenic marginal-zone leukemia, 3 (13%) with prolymphocytic leukemia (PLL) or PLL/CLL, and 1 (4%) with CLL. No cyclin D1 protein expression was noted by Western blot analysis in the one case that showed typical CLL morphology, and this patient did not require therapy. On the whole, the survival rate of CD23- patients was significantly worse than that of patients with CD23+. In contrast, 15 of 32 (49%) CD19+/CD5- patients were CD23-. CD23 negativity in this group was not associated with distinct clinical features or outcome. Eleven (73%) of these patients were classified as having splenic marginal-zone lymphoma and 4 as having follicular lymphoma. CONCLUSIONS: These data indicate that CD23 negativity is rare in typical B-cell CLL, and CD23 negativity in patients with CD19+/CD5+ is suggestive of mantle cell leukemia a more aggressive disease with poor response to conventional therapy in which newer chemotherapy regimens such as hyper-CVAD may be more effective. PMID- 11920536 TI - Comparison in treatments of nonleukemic granulocytic sarcoma: report of two cases and a review of 72 cases in the literature. AB - BACKGROUND: The purpose of this study was to reveal the clinical characteristics of nonleukemic granulocytic sarcoma (GS) and an association between the therapeutic regimens and the nonleukemic period. METHOD: Clinical records of 2 patients reported here and 72 patients gathered using a literature search on Medline from other institutions were analyzed. The patients consisted of 57 patients who preceded acute nonlymphoblastic leukemia (ANLL) and 17 patients who did not develop ANLL. These patients were divided into 3 groups by therapeutic regimens; Group I included 12 patients who received only biopsy or surgical resection of the tumor, Group II was 20 patients who received local irradiation for the tumor, and Group III consisted of 42 patients who received systemic chemotherapy. The nonleukemic periods between these groups were compared. In Group III, the period in the patients who were treated with chemotherapy given to ANLL was compared with that in the patients who received chemotherapy used for malignant lymphoproliferative disorders (MLPDs). RESULTS: Thirty-five patients (47%) initially were misdiagnosed, and the disease was most often malignant lymphoma. Preferential sites of GS were the small intestine, mediastinum, epidural site, uterus, and ovary, which often are difficult for the detection and diagnosis in addition to the skin and lymph nodes known commonly. The nonleukemic period after the diagnosis of GS was significantly longer in Group III than in the other groups (median, 12 months in Group III vs. 3 and 6 months in Groups I and II, respectively). The aggressive chemotherapy given to ANLL led to a longer nonleukemic period than the chemotherapy used for MLPDs. CONCLUSIONS: To reduce the risk of subsequent ANLL in patients with nonleukemic GS, it is important that accurate histologic diagnosis is established initially for GS and that all isolated cases of GS, even those that appear to be cured by resection or irradiation of the tumor, are treated with intensive chemotherapy similar to that used to treat ANLL during the nonleukemic period as soon as possible. PMID- 11920535 TI - European Organization for Research and Treatment of Cancer and Groupe d'Etude des Lymphomes de l'Adulte very favorable and favorable, lymphocyte-predominant Hodgkin disease. AB - BACKGROUND: Lymphocyte-predominant Hodgkin disease (LPHD) is rare and has a natural history different from that of classic Hodgkin disease. There is little information in the literature regarding the role of chemotherapy in patients with early-stage LPHD. The objective of this study was to examine recurrence free survival (RFS), overall survival (OS), and patterns of first recurrence in patients with LPHD who were treated with radiotherapy alone or with chemotherapy followed by radiotherapy. METHODS: From 1963 to 1996, 48 consecutive patients ages 16-49 years (median, 28 years) with Ann Arbor Stage I (n = 30 patients) or Stage II (n = 18 patients), very favorable (VF; n = 5 patients) or favorable (F; n = 43 patients) LPHD, according to the European Organization for Research and Treatment of Cancer and Groupe d'Etude des Lymphomes de l'Adulte (EORTC-GELA) criteria, received radiotherapy alone (n = 37 patients) or received chemotherapy followed by radiotherapy (n = 11 patients). The percentages of patients with VF disease (11% vs. 9% in the radiotherapy group vs. the chemotherapy plus radiotherapy group, respectively) or F disease (89% vs. 91%, respectively) within the two treatment groups were similar (P = 1.00). A median of three cycles of chemotherapy with mechlorethamine, vincristine, procarbazine, and prednisone (MOPP) or with mitoxantrone, vincristine, vinblastine, and prednisone (NOVP) was given initially to six patients and five patients, respectively. A median total radiotherapy dose of 40 grays (Gy) given in daily fractions of 2.0 Gy was delivered to both treatment groups. RESULTS: The median follow-up was 9.3 years, and 98% of patients were observed for > or = 3.0 years. RFS was similar for patients who were treated with radiotherapy alone and patients who were treated with chemotherapy followed by radiotherapy (10-year survival rates: 77% and 68%, respectively; P = 0.89). The OS rate also was similar for the two groups (10-year survival rates: 90% and 100%, respectively; P = 0.43). MOPP or NOVP chemotherapy did not reduce the risk of recurrence outside of the radiotherapy fields. CONCLUSIONS: MOPP or NOVP chemotherapy did not improve RFS or OS significantly in patients with VF or F LPHD, although the statistical power was limited. Ongoing clinical trials will help to clarify the role of a watch-and-wait strategy or systemic therapy, including anthracycline (epirubicin or doxorubicin), bleomycin, and vinblastine-based chemotherapy or antibody-based approaches, in the treatment of these patients. PMID- 11920537 TI - A prospective study regarding the complications of transcatheter intraarterial lipiodol chemoembolization in patients with hepatocellular carcinoma. AB - BACKGROUND: Hepatocellular carcinoma (HCC) is a common cause of cancer death throughout the world. The majority of patients are not suitable for curative resection either because of the advanced stage of the disease at the time of presentation or because of underlying cirrhosis. Transcatheter intraarterial lipiodol chemoembolization (TACE) has been reported to be one of the most effective palliative measures for HCC. However, its severe side effects continue to make its use controversial. METHODS: In the current study, the authors prospectively evaluated 197 sessions of TACE performed in 59 patients with HCC. RESULTS: Acute hepatic decompensation occurred in 20% of the 197 sessions with 3% of cases being irreversible. Significant elevation of bilirubin was associated with the dosage of cisplatin used (P = 0.0001), basal bilirubin level (P = 0.0001), basal prothrombin time (P =0.004), basal aspartate aminotransferase (AST) level (P = 0.013), and stage of cirrhosis (P < 0.0001). Patients with irreversible hepatic decompensation were more likely to have higher pre-TACE bilirubin levels (P = 0.009), more prolonged prothrombin time (P = 0.015), received a higher dose of cisplatin (P = 0.033), and more advanced cirrhosis (P < 0.0001). The majority of the other side effects were self-limiting with the exception of one patient who died of liver and splenic abscesses. Approximately 36% of the patients achieved a tumor response, 39% achieved stable disease, and 29% developed progressive disease. CONCLUSIONS: The results of the current study identified factors that appeared to predispose patients to irreversible hepatic decompensation after TACE. Despite the high percentage of patients who developed hepatic decompensation after TACE, irreversible damage occurred in only a minority. PMID- 11920538 TI - Echogenicity of liver metastases from colorectal carcinoma is an independent prognostic factor in patients treated with regional chemotherapy. AB - BACKGROUND: Echogenicity of liver metastases was found to be a predictive biologic factor influencing long-term outcome after curative liver resection. The current analysis focuses on the influence of echogenicity on survival in patients treated with intraarterial chemotherapy for unresectable colorectal carcinoma liver metastases. METHODS: A retrospective analysis of prospectively collected data at the Department of Surgery at the University of New South Wales-affiliated St. George Hospital was performed. Two hundred twelve consecutive patients with unresectable hepatic metastases from colorectal carcinoma treated between May 1992 and September 2000 were analyzed. Echogenicity of metastases was measured intraoperatively using a 5 MHz probe. Overall survival difference was compared between hyper- and hypoechoic metastases on an intention-to-treat basis. RESULTS: At a median followup of 15.1 months, 47 patients (22%) were alive and 165 (78%) had died. A significant survival benefit was observed in patients having hyperechoic lesions (median survival 16.2 months, 95% confidence interval [CI] 13.9-18.5) compared to hypoechoic lesions (median survival 11.6 months, 95% CI 8 15.2), P < 0.01. Other prognostic factors were differentiation of the primary tumor (P < 0.02), percentage hepatic replacement (P < 0.05) and carcinoembryonic antigen decrease (P < 0.03). Echogenicity was identified as an independent prognostic factor in multivariate analysis (P < 0.009). CONCLUSIONS: Echogenicity is an important prognostic survival parameter. PMID- 11920539 TI - Construction of the Chinese University Prognostic Index for hepatocellular carcinoma and comparison with the TNM staging system, the Okuda staging system, and the Cancer of the Liver Italian Program staging system: a study based on 926 patients. AB - BACKGROUND: The current TNM staging system for patients with hepatocellular carcinoma (HCC) does not include liver function parameters and does not provide a precise prognosis for patients in different risk groups. The objectives of this study were to construct a new prognostic index for patients with hepatocellular carcinoma, the Chinese University Prognostic Index (CUPI), and to compare it with existing staging systems in terms of their ability to classify patients into different risk group. METHODS: From 1996 to 1998, 926 ethnic Chinese patients who were diagnosed with HCC (mainly hepatitis B-associated) at a single institution were recruited prospectively into this study. A multivariate analysis on 19 patient characteristics was performed using a Cox regression model to identify independent prognostic factors. Weights were derived from the regression coefficients of various factors to construct the CUPI. Patients were classified according to different staging systems. Survival curves were plotted with the Kaplan-Meier method and were compared by using a log-rank test. RESULTS: Both the TNM staging system and the Okuda staging system had prognostic significance, but the significance was lower for the Cancer of the Liver Italian Program (CLIP) prognostic score among the patients in the study population. The CUPI was constructed by adding the following factors into the TNM staging system: total bilirubin, ascites, alkaline phosphatase, alpha fetoprotein, and asymptomatic disease on presentation. The new CUPI characterized three risk groups with highly significant differences in survival during the whole period of follow-up (P < 0.00001) and was more discriminant than the other systems. CONCLUSIONS: In the study population of patients with mainly hepatitis B-associated HCC, the CUPI was more discriminant than the TNM staging system, the Okuda staging systems, or the CLIP prognostic score in classifying patients into different risk groups and was better at predicting survival. The CUPI needs to be validated by different cohorts of patients before it can be recommended for general use. PMID- 11920540 TI - Correlation between expression of MUC1 core protein and outcome after surgery in mass-forming intrahepatic cholangiocarcinoma. AB - BACKGROUND: It has been reported that MUC1 is an important prognostic factor in several cancers. This study investigated the importance of MUC1 as a prognostic factor in mass-forming intrahepatic cholangiocarcinoma (m-ICC). METHODS: In 50 patients with m-ICC who had undergone hepatectomy, expression of MUC1 was investigated. Expression of MUC1 was examined by immunohistochemical staining with monoclonal antibody HMPV, which recognizes the MUC1 core peptide. The immunohistochemical staining patterns of MUC1 were classified into three types: ductal type (the luminal surface membrane of neoplastic cells was stained), cytoplasmic type (the cytoplasm of neoplastic cells was stained dominantly), and negative type. RESULTS: Expression of MUC1 was detected immunohistochemically in 38 (76%) of 50 cases of m-ICC (ductal type, 18; cytoplasmic type, 20; and negative type, 12). Seventy-five percent of patients with lymph node metastasis had the cytoplasmic type MUC1 expression. Lymph node dissection was performed in only 20 patients, but significant correlation was demonstrated between MUC1 expression and lymph node metastasis (P = 0.0227). The location of MUC1 expression correlated with surgical outcome in m-ICC. Patients with the cytoplasmic type expression showed significantly lower survival rates. Univariate analysis revealed that MUC1 expression was a statistically significant risk factor affecting outcome in m-ICC (P = 0.0028). Furthermore, expression of MUC1 was found to be a statistically significant independent risk factor in multivariate analysis (P = 0.0063). CONCLUSIONS: The results suggest that evaluation of MUC1 expression may be very useful in predicting the surgical outcome in m-ICC. PMID- 11920541 TI - Expression of smooth muscle calponin in tumor vessels of human hepatocellular carcinoma and its possible association with prognosis. AB - BACKGROUND: Hepatocellular carcinoma (HCC) is a vascular-rich tumor. The tumor vessels in HCC were demonstrated to have alpha-smooth muscle actin positive smooth muscle cells (SMCs). However, it is unclear whether the SMCs in the wall of the tumor vessels are differentiated or undifferentiated. Basic calponin is an actin-, tropomyosin-, and calmodulin-binding protein, and expression of the calponin gene in SMCs has been recognized as one of the late stage differentiation markers of SMCs. The authors investigated the differentiation state of SMCs in tumor vessels by immunohistochemical examination of calponin in patients with HCC, and whether it is associated with the patients' prognosis. METHODS: Tumor and nontumor tissues were obtained from 75 patients with HCC who underwent radical hepatic resection. The differentiation state of the smooth muscle cells were evaluated based on the expression level of calponin, an actin binding protein, using immunohistochemistry and reverse transcription-polymerase chain reaction analysis. The disease free survival (DFS) rates were estimated according to the Kaplan-Meier method comparing groups of patients with calponin positive and negative tumor vessels. A multivariate analysis based on the Cox proportional hazards regression model was performed to estimate whether the expression of calponin is an independent prognostic factor. RESULTS: In the 75 patients with HCC examined, 36 patients (48%) possessed calponin positive SMCs, and the remaining 39 (52%) did not. There were no significant differences in either clinical or pathologic factors between the two groups of patients. The 5- and 8-year DFS rate of the patients with calponin positive vessels were 37% and 26%, respectively. These values were significantly higher (11% and 5%) than those of patients with calponin negative vessels. Gender, TNM classification, perioperative transfusion, and calponin expression were found to be independent prognostic factors for DFS. CONCLUSIONS: Immunohistochemical examination of the calponin expression in the tumor vessels is a new and useful means to predict the prognosis of HCC patients after hepatic resection. PMID- 11920542 TI - Close association between high serum alanine aminotransferase levels and multicentric hepatocarcinogenesis in patients with hepatitis C virus-associated cirrhosis. AB - BACKGROUND: Multicentric development of hepatocellular carcinoma (HCC) is a characteristic feature of hepatitis C virus (HCV)-associated cirrhosis (HCV-LC). In this study, the objective was to determine whether the persistent elevation of the serum alanine aminotransferase (ALT) level, which represents the inflammatory necrosis of hepatocytes, is correlated with the multicentric development of hepatocellular carcinoma (HCC) in patients with early-stage HCV-LC. METHODS: Ninety-three consecutive patients with biopsy proven HCV-LC (Child Stage A) who had been followed for > 5 years for the development of HCC were studied. They were subdivided into three groups according to their serum ALT level: Group A included 33 patients with annual average serum ALT levels that were persistently high (> or = 80 IU; high ALT group), Group B included 41 patients with annual average serum ALT levels that were persistently low (< 80 IU; low ALT group), and Group C included 19 unclassified patients. The patients had been studied prospectively with frequent ultrasonography and magnetic resonance imaging or computed tomography (CT) scans for > 5 years. When the development of HCC was suspected, angiography, infusion of lipiodol into the hepatic artery, and lipiodol-CT scans were performed in all patients to determine the number of HCC nodules. RESULTS: In Group A, 27 patients (81.8%) developed HCC. Seventeen of 27 patients (63.0%) had multiple nodules. In contrast, in Group B, only 12 patients (29.3%) developed HCC, and only 1 of these 12 patients (8.3%) had multiple nodules. There was a significant difference between Groups A and B in the incidence of developing HCC (P < 0.001) and developing multiple nodules (P = 0.006). In addition, among the male patients, the incidence of developing multiple HCC nodules in Group A (12 of 19 patients; 63.2%) was significantly higher (P < 0.05) compared with the incidence in Group B (0 of 6 patients; 0%). The same tendency was observed among the female patients. CONCLUSIONS: These results showed a close correlation between multicentric hepatocarcinogenesis and sustained necroinflammation of the liver in patients with HCV-LC. PMID- 11920543 TI - 99mTC-tetrofosmin scintigraphy in lung carcinoma staging and follow-up evaluations. AB - BACKGROUND: 99mTC-tetrofosmin recently has emerged as a new radiopharmaceutical for cancer visualization. In this study, the authors have investigated, for the first time in a comprehensive way, its ability to assess lung carcinoma dissemination and progression. METHODS: A 99mTC-tetrofosmin scan was incorporated into the pretreatment and posttreatment diagnostic workup of all lung carcinoma patients seen in a second referral institution for a province of 500,000 inhabitants during the years 1998 and 1999. Sixty-one patients, strongly suspected of lung carcinoma were photon-scanned; 21 of them were rescanned after completion of their front-line treatment. Eleven patients eventually underwent surgery, and 3 others underwent mediastinoscopy. Both planar and single photoemission computed tomography thoracic views were obtained. Images for the whole body also were acquired. RESULTS: All 57 patients whose lung carcinoma was pathologically confirmed showed accumulation of the radiotracer (100% sensitivity). However, three of the four nonmalignant lesions were also 99mTC tetrofosmin positive. 99mTC-tetrofosmin scan was highly sensitive for the detection of the T0-T2 disease (97% sensitivity) and highly specific for the N0 N1 disease (83% specificity). In the 16 pathologically staged mediastina, sensitivity, specificity, and accuracy rates were 73%, 100%, and 81%, respectively. 99mTC-tetrofosmin scan correctly detected most skeleton (9 of 10) and brain (5 of 7) metastases. The treatment response evaluation made with 99mTC tetrofosmin corresponded to the clinical estimate in almost half of the sample. CONCLUSIONS: This study shows that 99mTC-tetrofosmin scan is a relatively accurate method for lung carcinoma evaluation. The authors' preliminary data exclude, however, that noninvasive diagnostic efficiency might be substantially increased by a scintigraphy with 99mTC-tetrofosmin. More studies are needed for a better understanding of the real value of this technique. PMID- 11920544 TI - Downregulation of Hus1 by antisense oligonucleotides enhances the sensitivity of human lung carcinoma cells to cisplatin. AB - BACKGROUND: In Schizosaccharomyces pombe, Hus1 is a component of the radiation sensitive (Rad) machinery that has been identified as playing a role in DNA repair and cell cycle G2/M checkpoint control pathways. Hus1 has been shown to exist in a discrete complex with at least two Rad family members, Rad1 and Rad9. Furthermore, Hus1 is essential for checkpoint activation, since Hus1 mutants fail to arrest the cell cycle in response to DNA damage or unreplicated DNA. To establish the role and relevance of human Hus1 in cell cycle regulation, the authors applied antisense technology to selectively downregulate the expression of Hus1 mRNA. METHODS: Transfection of 2'-O-methoxyethyl-modified Hus1 antisense oligoribonucleotides into human H1299 nonsmall lung carcinoma cells was performed using Lipofectin as the carrier. The authors prepared RNA from transfected cells, and levels of Hus1 expression were analyzed by real time polymerase chain reaction. The growth and viability of cells treated with Hus1 antisense oligonucleotides in the presence or absence of cisplatin were analyzed and compared to controls. RESULTS: Transfection of selected Hus1 antisense oligonucleotides into p53 deficient H1299 cells resulted in significant downregulation of Hus1 mRNA, up to 80%; RNA analyses reveal a maximal Hus1 antisense activity at a concentration of 200 nM with an IC50 determined to be 90 nM. The design and transfection of oligonucleotides containing three mismatches to their corresponding antisense counterparts had no or only minor effects on Hus1 mRNA levels, showing the specificity of Hus1 mRNA downregulation. The cisplatin IC50 in untransfected H1299 cells was found to be 20 microM and could be reduced significantly to only 7 microM after transfection of a Hus1 antisense oligonucleotide. CONCLUSIONS: Experiments addressing the proliferation and viability of transfected H1299 cells suggest that downregulation of Hus1 by specific antisense oligonucleotides sensitizes human cells to treatment with the DNA damaging agent cisplatin. PMID- 11920545 TI - Cerebrospinal fluid concentrations of vincristine after bolus intravenous dosing: a surrogate marker of brain penetration. AB - BACKGROUND: Vincristine (VCR) is used widely in oncology practice, and regular dosing is commonly associated with the development of sensorimotor or autonomic neuropathies. However, the incidence of VCR-related central nervous system (CNS) toxicity is comparatively low, suggesting that the blood-brain barrier may limit drug penetration into the brain parenchyma. This study determined whether measurable concentrations of VCR could be detected in the cerebrospinal fluid (CSF), as a surrogate marker of brain parenchyma penetration, after bolus intravenous injection in children without primary CNS pathology. METHODS: The authors studied 17 pediatric patients ages 2.5-14.1 years (median, 6.8 years) with acute lymphoblastic leukemia or non-Hodgkin lymphoma without evidence of leptomeningeal disease. Patients received VCR 1.5 mg/m2 by intravenous bolus injection followed at varying intervals by lumbar puncture for scheduled intrathecal methotrexate administration under general anesthesia. Paired VCR concentrations in both plasma and CSF were measured in each patient simultaneously at times ranging from 8 minutes to 146 minutes after the VCR injection. Three patients were studied twice. The paired samples were stored at 40 degrees C until analysis using a high performance liquid chromatography assay with a sensitivity of 0.1 microg/L in CSF and 0.4 microg/L in plasma. RESULTS: Plasma VCR concentrations ranged from 2.2 microg/L to 91.2 microg/L. No measurable VCR concentrations were detected in the CSF samples. CONCLUSIONS: Measurable concentrations of VCR in CSF are not achieved after the administration of standard intravenous bolus doses of VCR. The current observations are consistent with the relative rarity of VCR-related CNS neurotoxicity compared with the commonly observed sensorimotor and autonomic neuropathies. These findings suggest that the penetration of VCR into the brain parenchyma of patients with a relatively intact blood-brain barrier is low and that VCR may have a limited role in the CNS-directed therapy of these patients. PMID- 11920546 TI - Understanding cancer metastasis: an urgent need for using differential gene expression analysis. AB - Cancer is a multistep process and occurs as a result of the loss of control of cell division, leading to the initial tumor formation, which is followed by metastatic spread. Recent years have witnessed a vast improvement in the understanding of the molecular mechanisms regulating cell division and their links to tumorigenesis. The process of metastasis involves an intricate interplay between cell adhesion, proteolysis, migration, and angiogenesis. However, there is little knowledge of how these events are coordinately regulated in the tumor cell. Given that the uncontrolled spread of the tumor to distant organs is usually lethal, a study of the molecular mechanisms regulating metastasis assumes great significance. Recently, several technologies have been developed for analyzing differential gene expression. The current review discusses the importance of these technologies in the molecular analyses of metastasis. PMID- 11920547 TI - A population-based study of pediatric anaplastic large cell lymphoma. AB - BACKGROUND: Anaplastic large cell lymphoma (ALCL) is a relatively rare form of non-Hodgkin lymphoma (NHL) in children constituting 10-15% of this entity. To the authors' knowledge, there are no population-based pediatric studies in the literature, and incidence estimates have not been attempted. METHODS: A population-based study of all children in British Columbia, Canada, younger than 15 years of age presenting between January 1988 and December 1999 with an immunohistologically confirmed diagnosis of CD30+ ALCL was completed. Demographic, ethnic, clinical, treatment, and outcome details were collected on all patients. Population figures were obtained from census data through the BC STATS. RESULTS: Ten patients were identified and confirmed to have ALCL on the basis of morphology and immunohistochemical stains. This equates to an annual incidence of 1.2 per million children younger than 15 years of age. The mean age at diagnosis was 8.23 years with a range of 1.4-13.0 years. There was an overrepresentation of East and Southeast Asian patients (40%) in the ALCL group compared with other subtypes of NHL and the pediatric population of British Columbia. Twenty percent of the patients had evidence of central nervous system (CNS) disease at diagnosis. CONCLUSIONS: The annual incidence of ALCL in children younger than 15 years of age in British Columbia is 1.2 per million. The overrepresentation of East and Southeast Asian ethnic origin remains unexplained. The rate of CNS involvement is much higher than that previously reported in large non-population-based series (0-3%). Although the low numbers do not allow firm conclusions, it would seem prudent to continue to investigate all newly diagnosed patients for CNS involvement in British Columbia. PMID- 11920548 TI - The panorama of opioid-related cognitive dysfunction in patients with cancer: a critical literature appraisal. AB - BACKGROUND: Opioids have an essential role in the management of pain in cancer patients, particularly those with advanced disease. Cognitive dysfunction is a recognized complication of opioid use. However, misconceptions and controversy surround the nature and prevalence of its occurrence. A projected increase in the aging cancer population highlights the need for a better understanding of this phenomenon. METHODS: A critical appraisal of the literature evidence in relation to the pattern, pathophysiology, assessment, impact, and management of cognitive dysfunction due to opioid use in cancer pain management is given. RESULTS: Studies in cancer patients with less advanced disease reveal subtle evidence of cognitive impairment, largely related to initial dosing or dose increases. In advanced cancer, opioid-induced cognitive dysfunction usually occurs in the form of delirium, a multifactorial syndrome. The presence of both cognitive impairment and delirium frequently is misdiagnosed or missed. Potential risk factors include neuropathic and incidental pain, opioid tolerance, somatization of psychologic distress, and a history of drug or alcohol abuse. Elevation of opioid metabolites with renal impairment may contribute to cognitive dysfunction. Recognition of opioid-related cognitive dysfunction is improved by objective screening. Successful management requires either dose reduction or a change of opioid, in addition to addressing other reversible precipitants such as dehydration or volume depletion. CONCLUSIONS: Opioid-related cognitive dysfunction tends to be subtle in the earlier stages of cancer, whereas delirium, a more florid form with behavioral disturbance is likely to be present in the advanced cancer population. In patients with advanced disease, an optimal management approach requires careful clinical assessment, identification of risk factors, objective monitoring of cognition, maintenance of adequate hydration, and either dose reduction or switching to a different opioid. PMID- 11920549 TI - Helping patients with localized prostate carcinoma manage uncertainty and treatment side effects: nurse-delivered psychoeducational intervention over the telephone. AB - BACKGROUND: The objective of this study was to test the efficacy of an individualized uncertainty management intervention delivered by telephone to Caucasian and African-American men with localized prostate carcinoma and directed at managing the uncertainties of their disease and treatment. METHODS: The authors delivered a psychoeducational intervention by phone to men with prostate carcinoma, with or without supplemented delivery to a close family member, that was directed at managing uncertainty and improving symptom control. One hundred thirty-four Caucasian men and 105 African-American men were assigned randomly to one of two approaches to delivering the intervention or to the control condition. Men entered the study immediately after surgical treatment or in the first 3 weeks of radiation therapy. Trained nurses delivered the intervention through weekly phone calls for 8 weeks. RESULTS: The authors found that the majority of intervention effects were from baseline to 4 months postbaseline, when treatment side effects are most intense. Both Caucasian men and African-American men who received either one of the two approaches for delivering the intervention improved in the two uncertainty management methods of cognitive reframing and problem solving. Similarly, when the intervention groups were combined, men who received the intervention also improved significantly in control of incontinence by 4 months postbaseline. Decreases in the number of treatment side effects differed by time and treatment/ ethnic group interactions as did satisfaction with sexual functioning. CONCLUSIONS: This is one of the first tests of a psychoeducational intervention among men with prostate carcinoma and was the first test that included a sufficient number of African-American men to test by ethnic group. Therefore, replication of these findings is advised. PMID- 11920550 TI - An estimate of premature cancer mortality in the U.S. due to inadequate doses of solar ultraviolet-B radiation. AB - BACKGROUND: There are large geographic gradients in mortality rates for a number of cancers in the U.S. (e.g., rates are approximately twice as high in the northeast compared with the southwest). Risk factors such as diet fail to explain this variation. Previous studies have demonstrated that the geographic distributions for five types of cancer are related inversely to solar radiation. The purpose of the current study was to determine how many types of cancer are affected by solar radiation and how many premature deaths from cancer occur due to insufficient ultraviolet (UV)-B radiation. METHODS: UV-B data for July 1992 and cancer mortality rates in the U.S. for between 1970-1994 were analyzed in an ecologic study. RESULTS: The findings of the current study confirm previous results that solar UV-B radiation is associated with reduced risk of cancer of the breast, colon, ovary, and prostate as well as non-Hodgkin lymphoma. Eight additional malignancies were found to exhibit an inverse correlation between mortality rates and UV-B radiation: bladder, esophageal, kidney, lung, pancreatic, rectal, stomach, and corpus uteri. The annual number of premature deaths from cancer due to lower UV-B exposures was 21,700 (95% confidence interval [95% CI], 20,400-23,400) for white Americans, 1400 (95% CI, 1100-1600) for black Americans, and 500 (95% CI, 400-600) for Asian Americans and other minorities. CONCLUSIONS: The results of the current study demonstrate that much of the geographic variation in cancer mortality rates in the U.S. can be attributed to variations in solar UV-B radiation exposure. Thus, many lives could be extended through increased careful exposure to solar UV-B radiation and more safely, vitamin D3 supplementation, especially in nonsummer months. PMID- 11920551 TI - Utilization of BRCA1/2 genetic testing in the clinical setting: report from a single institution. AB - BACKGROUND: Clinical testing for BRCA1/2 has been available since 1996. Interest in testing in the research and hypothetical situations has been consistently high, but there have been limited reports on its clinical utilization. METHODS: This is a retrospective study of BRCA1/2 test utilization by high-risk patients who were seen at the Johns Hopkins Breast and Ovarian Surveillance Service. RESULTS: Between February 1996 and December 1999, 258 families who had at least a 10% chance of carrying a BRCA1/2 mutation were offered genetic testing. Of these, 26 families seen between February 1996 and October 1996 had access to free testing. Overall, 68 of 258 (26%) underwent genetic testing. Educational level, number of children or daughters, breast carcinoma screening behavior, smoking and drinking behavior, perceived risk of breast carcinoma, and family history was not associated with test utilization. Eligibility for free testing, prior history of breast or ovarian carcinoma, Ashkenazi Jewish versus non-Ashkenazi Jewish heritage, genetic risk category, and age category were associated with test utilization, and in multivariate analysis, the first three remained statistically significant factors associated with genetic testing. Only 26% of the 50 patients who did not have access to free testing sought insurance reimbursement, of which greater than 50% (7 of 13) had a prior diagnosis of breast or ovarian carcinoma. CONCLUSIONS: The actual utilization of BRCA1/2 genetic testing in a clinical setting is lower than in the research and hypothetical settings. Potential obstacles include cost, fear of insurance discrimination, and need to involve an affected family member in the testing process. PMID- 11920552 TI - Ethnic differences in survival among women with ovarian carcinoma. AB - BACKGROUND: Ovarian carcinoma is the leading cause of death among all female reproductive malignancies. There are substantial differences in age-adjusted incidence rates and survival rates between Caucasian women and African-American women. The objective of this study was to examine ethnic differences in survival after ovarian carcinoma in a population-based sample of women. METHODS: Thirteen thousand eighty-three patients (12285 Caucasian women and 798 African-American women) who were diagnosed with primary ovarian carcinoma from the population based Surveillance, Epidemiology, and End Results (SEER) Program were used for analysis. Odds ratios were used to estimate the association between prognostic variables and ethnicity. Chi-square tests were used to determine the statistical significance of these associations (using two-sided P values). Univariable and multivariable Cox proportional hazards models were used to assess survival differences. RESULTS: African-American women were significantly younger at the time of diagnosis, were more likely to be single, and were less likely to undergo site specific surgery compared with Caucasian women. In addition, the crude median survival for African-American women was nearly 1 year less than for Caucasian women (22 months vs. 32 months, respectively; P < 0.0001). African American women were at a 30% increased risk of death from any cause when adjusting for all other prognostic variables that differed between the two ethnic groups. CONCLUSIONS: African-American women who are diagnosed with ovarian carcinoma are at a significant increased risk of death from any cause compared with Caucasian women who are diagnosed with ovarian carcinoma. PMID- 11920553 TI - Cytoplasmic expression of laminin gamma2 chain correlates with postoperative hepatic metastasis and poor prognosis in patients with pancreatic ductal adenocarcinoma. AB - BACKGROUND: The laminin gamma2 chain is involved in tumor invasion and metastasis, but the significance of laminin gamma2 chain expression remains unclear in patients with pancreatic carcinoma. METHODS: Laminin gamma2 chain expression was examined immunohistochemically in 48 patients with pancreatic ductal adenocarcinoma who were followed closely to elucidate the correlations between clinicopathologic factors, postoperative recurrence, and overall survival. Prognostic factors for postoperative survival were examined comparing clinicopathologic factors and laminin gamma2 chain expression. RESULTS: Two different staining patterns of laminin gamma2 chain expression, cytoplasmic expression and basement membrane expression, were detected in tumors from all 48 patients. Tumors were then classified into two types according to the dominant pattern of laminin gamma2 chain expression: the cytoplasmic expression dominant type (CYT; n = 26 patients) and the basement membrane expression dominant type (BM; n = 22 patients). Tumor differentiation was associated statistically with the BM type of laminin gamma2 chain expression (P = 0.0002). The CYT type of laminin gamma2 chain expression was associated significantly with the occurrence of postoperative hepatic metastasis (P = 0.0011) and also was the strongest predictive factor for poorer overall survival in patients with pancreatic ductal adenocarcinomas (P = 0.0161). CONCLUSIONS: The cytoplasmic expression of the laminin gamma2 chain represents the high invasive potential of the tumor and is correlated with distant metastasis, especially hepatic metastasis, and with a poorer prognosis in patients with pancreatic ductal adenocarcinoma. PMID- 11920554 TI - Time trends of cutaneous melanoma in Queensland, Australia and Central Europe. PMID- 11920555 TI - Increased expression of cyclooxygenase-2 in human pancreatic neoplasms and potential for chemoprevention by cyclooxygenase inhibitors. PMID- 11920556 TI - Breast-conserving therapy for Paget disease of the nipple: a prospective European Organization for Research and Treatment of Cancer study of 61 patients. PMID- 11920557 TI - Plasma cells for a lifetime? AB - Antigen-specific serum antibodies are protective for long periods of time 1, 2. These serum antibodies, the "humoral memory", are secreted by plasma cells derived from activated, antigen-specific B lymphocytes. Given their crucial role in immunity, surprisingly little is known about the biology of plasma cells. One of the fundamental questions is whether persisting protective serum antibody responses are maintained by long-lived plasma cells, or by short-lived plasma cells generated continuously from activated memory B cells. Here, we review some recent experiments suggesting that plasma cells have the capacity to live for unlimited time if rescued by specific factors provided in a limited number of survival niches in the body. PMID- 11920558 TI - Mechanism of murine Vgamma1+ gamma delta T cell-mediated innate immune response against Listeria monocytogenes infection. AB - Murine gamma delta T cells participate in innate immune response against infection of the intracellular bacterium Listeria monocytogenes. In the present report, we analyzed the mechanism of the gamma delta T cell-mediated response against L. monocytogenes infection. gamma delta T cell-enriched spleen cells of L. monocytogenes-infected mice produced IFN-gamma in vitro in response to L. monocytogenes-infected spleen cells. The IFN-gamma production was abrogated by depletion of Vgamma1+ gamma delta T cells. IFN-gamma production of the Vgamma1+ gamma delta T cells in response to L. monocytogenes-infected spleen cells required IL-12. However, addition of Fab fragment of anti-TCR gamma delta monoclonal antibodies (mAb) failed to block the response, suggesting that the response requires no TCR-mediated antigen recognition. Interestingly, Vgamma1+ gamma delta T cells of naive mice also produced IFN-gamma in response to L. monocytogenes-infected spleen cells in an IL-12-dependent manner. Furthermore, the IL-12 receptor (IL-12R) gene was expressed on the Vgamma1+ gamma delta T cells of naive mice as well as those of L. monocytogenes-infected mice although naive alpha beta T cells lack IL-12R expression. All the results suggest that the Vgamma1+ gamma delta T cells participate in immune surveillance against intracellular bacterial infection through quick production of IFN-gamma in response to infection-induced IL-12 without antigen-driven clonal expansion and maturation. PMID- 11920559 TI - Human T cell receptor-mediated recognition of HLA-E. AB - The HLA-E class Ib molecule presents hydrophobic peptides derived from the leader sequences of other class I molecules, constituting the ligands for CD94/NKG2 lectin-like receptors. Along the course of our studies on human CD94+ T cells, we characterized an alpha beta CD8+CD94/NKG2C+ CTL clone (K14). In cytolytic assays against the murine TAP-deficient RMA-S cells transfected with human beta2 microglobulin and HLA-E (RMA-S/HLA-E), loaded with different synthetic peptides, K14 displayed a pattern of specific recognition distinct to that observed in CD94/NKG2C+ NK clones tested in parallel. RMA-S/HLA-E cells loaded with some but not all HLA class I leader sequence peptides were efficiently recognized by K14 but not by CD94/NKG2C clones, andvice versa. Remarkably, K14 also reacted with HLA-E loaded with a peptide derived from the BZLF-1 Epstein-Barr virus protein. Anti-CD94 mAb did not prevent K14 cytotoxicity against RMA-S/HLA-E cells, whereas incubation with anti-clonotypic mAb specific for the K14 TCR markedly inhibited lysis. Soluble HLA-E tetramers refolded with different peptides (i.e. VMAPRTVLL, VMAPRTLIL, VMAPRTLFL) specifically stained K14 cells. HLA-E tetramer binding was minimally reduced by pretreatment with anti-CD94 mAb alone, but was completely prevented in combination with anti-clonotypic mAb. Altogether, the data unequivocally imply the generation of human T cells potentially recognizing through the alpha beta TCR HLA-E molecules that bind to class I- and virus derived peptides. PMID- 11920560 TI - Neonatal infection with a milk-borne virus is independent of beta7 integrin- and L-selectin-expressing lymphocytes. AB - Mouse mammary tumor virus (MMTV) is acquired by neonates through milk and first infects lymphocytes in Peyer's patches. We show here that newborn mice lacking beta7 integrin or L-selectin were infected with MMTV at wild-type levels in both their lymphoid and mammary tissues. Superantigen-mediated activation and cognate T cell deletion were also unimpaired in both types of null mice. A large proportion of neonatal Peyer's patch lymphocytes in wild-type mice were beta7 and beta1 integrin low and both populations increased in response to MMTV infection. These results suggest that adhesion molecules other than beta7 integrin or L selectin play a role in lymphocyte homing in the gut, peripheral lymph nodes and mammary gland in response to MMTV infection. PMID- 11920561 TI - Contribution of light chain rearrangement in peripheral B cells to the generation of high-affinity antibodies. AB - Recently, peripheral B cells have been shown to undergo secondary V(D)J rearrangement of immunoglobulin genes, but the physiological role of this event has not been fully elucidated. To investigate whether rearrangement of L chain genes in the periphery is involved in the generation of high-affinity antibodies (Ab), we used the 17.2.25 rearranged VHDJH gene (VHT)-knockin mouse whose B cell diversity is limited due to the expression of the site-directed transgene. Immunization of the mouse with p-nitrophenylacetyl (pNP)-conjugated chicken gamma globulin preferentially led to the production of anti-pNP IgG Ab comprised of non VHT-encoded H chains and lambda chains. lambda(+) IgG constituted a majority of high-affinity Ab to this hapten. RAG-2 mRNA and the recombination signal sequence break of the lambda1 gene increased in the draining lymph node of immunized mice, but not of nonimmunized animals. There was a close correlation between the levels of these parameters implicating lambda gene rearrangement and the production of lambda(+ )high-affinity anti-pNP IgG. These observations were reproduced in RAG-1 deficient mice that were reconstituted with the spleen cells ofthe knockin mouse. Thus, our findings suggest that L chain rearrangement that occurs in the periphery can contribute to affinity maturation of Ab. PMID- 11920562 TI - Wnt-1 and Wnt-4 regulate thymic cellularity. AB - Thymic primordium, formed by cells derived from the endoderm, the ectoderm and the neural crest-derived mesenchyme, receive fetal liver derived lymphoid precursors. Reciprocal cell-cell interactions between thymic stromal cells and lymphoid precursors are critical in the expansion and maturation of thymocytes. Transcription factor TCF-1 is critical for the expansion of thymocytes because deletion of TCF-1 results in a significant decrease in the number of thymocytes without affecting the developmental pattern. In this report we show that Wnt-1 and Wnt-4 are expressed in the thymus and the deletion of Wnt-1 or Wnt-4 result in a substantial decrease in the number of thymocytes without affecting the pattern of maturation. Wnt-1 and Wnt-4 both regulate developing thymocytes because a double deficiency results in a significantly greater decrease of immature and mature thymocytes compared to deficiency in either Wnt-1 or Wnt-4. PMID- 11920563 TI - The role of CTLA-4 in induction and maintenance of peripheral T cell tolerance. AB - T cell receptor engagement and the B7-CD28 / CTLA-4 signaling pathways play critical roles in T cell activation and regulation. CD28 engagement results in T cell activation, differentiation and survival while CTLA-4 signals block IL-2 production, cell cycle progression and T cell differentiation. We explored the role of CTLA-4 in peripheral tolerance induced by intravenous administration of ethylene carbodiimide-fixed, antigen-coupled splenocytes in the PLP139 - 151 induced relapsing experimental autoimmune encephalomyelitis system. Tolerance induction with PLP139 - 151-coupled splenocytes correlates with low B7 expression on the fixed antigen-presenting cells, conditions that would favor CTLA-4 mediated inhibition. Administration of CTLA-4Ig or anti-CTLA-4 concomitant with the 'tolerogenic' stimulus, however, failed to reverse tolerance induction. In contrast, blocking CTLA-4 at the time of secondary 'immunogenic' encounter with antigen reversed the tolerant state. These findings indicate that CTLA-4 is required to maintain the unresponsive state of the tolerized T cells upon antigenic stimulation under inflammatory conditions and, therefore, have important implications for therapeutic regulation of autoimmune disease. PMID- 11920564 TI - Expression profiling in transformed human B cells: influence of Btk mutations and comparison to B cell lymphomas using filter and oligonucleotide arrays. AB - We have used both Clontech Atlas Human Hematology/Immunology cDNA microarrays, containing 588 genes, and Affymetrix oligonucleotide U95Av2 human array complementary to more than 12,500 genes to get a global view of genes expressed in Epstein-Barr virus (EBV)-transformed B cells and genes regulated by Bruton's tyrosine kinase (Btk). We compared EBV-transformed wild-type (WT) B cells from a healthy individual, WT1 and an X-linked agammaglobulinemia (XLA) patient cell line, XLA1, using the Clontech filters arrays. Eleven genes were > or =1.9-fold induced in absence of functional Btk. Furthermore, we analyzed a second patient cell line, XLA2, and compared this to two WT cell lines using oligonucleotide arrays. A total of 391 genes were found to be differentially expressed, including kinases and transcriptions factors. Furthermore, one expressed sequence tag and eight complementary DNA clones with unknown function were down-regulated in XLA2, indicating their biological role. Higher-fold inductions, Fyn (39.5), Hck (15.5) and Cyp1B1 (5.8), were observed using oligonucleotide array and were confirmed using real-time PCR for Fyn (20.8), Hck (6.7) and Cyp1B1 (10). Two genes, B cell translocation gene1 (BTG1) and B cell-specific OCT binding factor-1 (OBF-1) were induced > or =1.9-fold in both XLA1 and XLA2 analyzed by Atlas filter arrays andAffymetrix chips, respectively. Data from both filter and oligonucleotide arrays were compared to the gene clusters of a previously published lymphoma expression profile by linking to the UniGene transcript database. Our findings demonstrate for the first time the use of microarray to study the influence of Btk mutations and the use of functional annotation and validation of expression data by comparison of microarray analyses. PMID- 11920565 TI - Autocrine IL-10 impairs dendritic cell (DC)-derived immune responses to mycobacterial infection by suppressing DC trafficking to draining lymph nodes and local IL-12 production. AB - The production of IL-12 by dendritic cells (DC) early in an immune response is considered critical for the polarization of CD4(+) T lymphocyte response towards a Th1 pattern, a key process in the clearance of intracellular pathogens. Infection of bone marrow-derived DC with Mycobacterium bovis Bacillus Calmette Guerin (BCG) induced a concurrent and dose-dependent releaseof IL-10 and IL-12. Here we examined whether the production of IL-10 by DC affected their IL-12 response to mycobacterial infection and the generation of protective immune responses in vivo. Compared to wild-type (WT) DC, DC deficient for IL-10 synthesis (IL-10(-/-)) showed increased IL-12 production in response to BCG infection and CD40 stimuli in vitro. Moreover, when transferred into mice, infected IL-10(-/-) DC were more efficient than WT DC at inducing IFN-gamma production to mycobacterial antigens in the draining lymph nodes (DLN). This effect was associated with increased trafficking of IL-10(-/-) DC to the DLN and enhanced IL-12 production by DC within the DLN. These data show that autocrine IL 10 exerts a dual inhibitory effect on the induction of primary immune responses by DC: first, by down-regulating the migration of infected DC to the DLN and second, by modulating the IL-12 production by DC in the DLN. PMID- 11920566 TI - Cruzipain, a major Trypanosoma cruzi antigen, conditions the host immune response in favor of parasite. AB - We recently demonstrated that humoral immune response to cruzipain, a major antigen of Trypanosoma cruzi parasite, is implicated in the pathogenesis of experimental Chagas' disease. In the present study, the spleen cell phenotype and the cytokine profile induced by cruzipain in immunized mice were analyzed. The results showed that cruzipain increases the number of spleen cells with large size and granularity. Splenocyte populations with CD19(+), Mac-1(+), Gr-1(+) and CD11c(+) positive surface markers significantly increased in immune mice compared to controls ones. Histological study revealed the presence of high number of megacariocyte and granulocyte-macrophage progenitors, indicating extramedullary hemopoiesis in spleens of immune mice. The finding of high levels of IL-4, IL5 and IL-10 and low levels of IFN-gamma and IL-12 in supernatants of immune cells stimulated with cruzipain indicates a preferential activation of T2 type cells in immune animals. To investigate the role of innate immunity cells, the classical and alternative metabolic pathways of spleen macrophages from immune mice stimulated by cruzipain were also studied. The results showed an increase of urea associated with a decrease of nitrite levels, suggesting that cruzipain up regulates the arginase way. Therefore, cruzipain leads to T2 type cytokine profile which may enhance the arginase via in the macrophages promoting a susceptible mechanism to infection. Thus, we postulate that during T. cruzi infection, cruzipain could be used by the parasite to spread inside the host. PMID- 11920567 TI - Chondroitin sulfate A released from platelets blocks RANTES presentation on cell surfaces and RANTES-dependent firm adhesion of leukocytes. AB - The sequestration of chemokines on the surface of microvascular endothelium is an early event in the selective recruitment of leukocytes. The sequestration and presentation of chemokines must be tightly controlled to confine the extravasation of leukocytes and to prevent uncontrolled inflammation. We investigated whether soluble molecules released under physiological conditions could control chemokine immobilization on cell surfaces and function as regulatory chemokine binding molecules. We determined that human serum contains a molecule that suppresses RANTES (CCL5) binding to endothelial cells, PBMC and CHO cells. Using platelet-rich and platelet-free plasma, serum from patients with thrombocytopenia, and purified platelets, we identified platelets as the source of the chemokine-binding molecule and further identified it as chondroitin sulfate A. In contrast to platelet-derived fully-sulfated chondroitin sulfate A, low-sulfated chondroitin sulfate A present in plasma was almost inactive. Under physiological flow conditions chondroitin sulfate A was found to block RANTES mediated firm adhesion of monocytes to endothelial cells. It also prevented RANTES-mediated influx of calcium in CCR5-transfected CHO cells while internalization of CCR5 was only marginally reduced. Taken together, chondroitin sulfate A released from platelets appears to act as an important regulatory molecule for cellular responses to chemokines. PMID- 11920568 TI - Autoimmune thyroid disease induced by thyroglobulin and lipopolysaccharide is inhibited by soluble TNF receptor type I. AB - Experimental autoimmune thyroiditis (EAT) is inducible in mice by immunization with thyroglobulin and adjuvant. Previous studies have shown that EAT is an autoimmune Th1-mediated disease but its characteristics differ with the adjuvant. Granulomatous lesions with marked follicular disruption develop following administration of thyroglobulin (Tg) and complete Freund's adjuvant (CFA) whereas when lipopolysaccharide (LPS) is used as the adjuvant only focal infiltrates of mononuclear cells are observed. The pro-inflammatory cytokine, TNF-alpha, is associated with Th1 autoimmune-mediated conditions. Cytokine antagonists have been used as potential therapeutic agents in several experimental autoimmune models. Soluble cytokine receptors belong to this category and may naturally be shed from cell membranes to inhibit cytokine activity. We show that the administration of the soluble TNF receptor type I (sTNFR I) in the induction of EAT has very different effects on the two models of induced autoimmune thyroiditis. sTNFR I treatment inhibits the induction of EAT only when mouse Tg is given with LPS not with CFA, suggesting an important difference in the pathogenic processes. PMID- 11920569 TI - The tyrosine kinase Lyn is required for B cell development beyond the T1 stage in the spleen: rescue by over-expression of Bcl-2. AB - We have analyzed the effects of deficiency in the tyrosine kinase Lyn on B cell development using transgenic mice that express a B cell antigen receptor (BCR) of defined specificity (3-83,anti-H-2K(k or b)). In the absence of Lyn, immature B cells are abundant in the bone marrow and spleen up until the T1 stage (IgM(hi) IgD(-) CD21(-)CD23(-)), after which B cell development is severely impaired. The small number of mature B cells that do develop in Lyn-deficient mice express normal levels of the transgenic BCR and lack expression of CD80 and CD86, suggesting they are not activated. In Lyn-deficient animals the presence of a Bcl 2 transgene leads to a dramatic increase in B cell numbers and restores T2 stage (IgM(hi) IgD(hi) CD21(hi) CD23(int)) and mature populations. In 3-83 lyn-/- Bcl-2 Tg mice, a population of lambda-positive cells that also express the 383 idiotype is evident, suggesting that in the absence of lyn isotype exclusion by the transgenic BCR is less efficient. The results indicate that Lyn plays a positive role in the selection and survival of mature B cells in addition to its previously documented negative role in tolerance and B cell activation. PMID- 11920570 TI - Killing of naive T cells by CD95L-transfected dendritic cells (DC): in vivo study using killer DC-DC hybrids and CD4(+) T cells from DO11.10 mice. AB - Dendritic cells (DC) play the dual task of initiating cellular immunity against potentially harmful foreign antigens (Ag), while maintaining immunological tolerance to self-Ag and environmental Ag. As an approach to induce Ag-specific suppression, we and others introduced CD95 ligand (L) cDNA into DC. The resulting "killer" DC delivered apoptotic signals, instead of activation signals, to primed CD4(+) T cells in vitro and induced Ag-specific immunosuppression in vivo. To study the impact of killer DC on naive T cells, the fate of Ag-reactive T cells and the extent of their depletion after killer DC treatment, we performed in vitro and in vivo reconstitution experiments using: (a) killer DC-DC hybrids created between CD95L-transduced XS106 DC clone (A/J origin) and splenic DC from BALB/c mice, (b) CD4(+) T cells isolated from DO11.10 transgenic mice (BALB/c background), and (c) OVA(323-339) peptide as relevant Ag. Ovalbumin (OVA)-pulsed killer DC-DC hybrids inhibited DO11.10 T cell activation triggered by conventional DC, instead of inducing their activation. Rapid apoptosis of T cells was observed after co-culture with OVA-pulsed killer DC-DC hybrids, but not with non-pulsed killer DC-DC hybrids or OVA-pulsed control DC-DC hybrids. For in vivo reconstitution, (BALB/cxA/J)F1 mice received subcutaneous administration of killer DC-DC hybrids, followed by intravenous inoculation of DO11.10 T cells. Killer DC-DC hybrids migrated preferentially to draining lymph nodes albeit with relatively low efficiency (0.5-1% recovery) and they induced significant, but incomplete (30-40%) killing of DO11.10 T cells in this location. These results document the abilities of CD95L-transduced DC to trigger apoptosis of naive T cells in an Ag-specific manner, to overrule T cell activation signals delivered by conventional DC, and to reduce local frequencies of Ag-reactive T cells in vivo. Our data also uncover two major limitations (relatively low homing efficiency and incomplete elimination of Ag-reactive T cells) that remain to be overcome for clinical application of CD95L-transduced DC strategy. PMID- 11920571 TI - Constitutive, but not inflammatory, cross-presentation is disabled in the pancreas of young mice. AB - Peripheral antigens can be captured by APC and cross-presented to naive CD8(+) T cells. Notably, cross-presentation of pancreatic antigen is not seen in neonatal mice, although presentation of antigen expressed by the kidney is still intact. In this report, we examined why pancreatic antigens are not cross-presented in neonatal mice. First, we established that antigen expression was not limiting, as neonatal islets expressed as much antigen per cell as adult islets, and vastly more than neonatal renal cells. Next, we analyzed the APC subsets present in the lymph node draining the neonatal pancreas. No obvious population was absent. Finally, we examined whether cross-presentation occurred during inflammation. This showed that inflammation caused by CTL attack of islet tissue facilitated cross-presentation of antigens in neonatal mice. These data indicate that constitutive cross-presentation of islet antigens is inactive during neonatal life, but that under inflammatory conditions this antigen presentation pathway becomes available. PMID- 11920572 TI - Alanine scanning mutagenesis of CCR3 reveals that the three intracellular loops are essential for functional receptor expression. AB - Intracellular signaling mediated by the eotaxin receptor, CCR3, has been implicated in allergic diseases involving the recruitment and activation of eosinophils. To investigate the structural requirements of the three intracellular loops (ICL) of CCR3, a panel of 15 alanine triplet mutants were generated and their effects on function assessed by assays of cell surface expression and chemotactic responsiveness. While the majority of constructs were efficiently expressed when compared with their wild-type counterpart, their abilities to migrate in response to eotaxin were relatively poor, suggesting that all three intracellular loops of CCR3 are involved to some degree in coupling to G proteins. - Another panel of 7 point mutants were then constructed to examine the DRY motif which resides in ICL2 and is highly conserved throughout the chemokine receptors identified to date. The conservative mutants D130E and R131K were well tolerated and gave chemotactic responses approaching 35 % of wild-type CCR3, but the less conserved substitutions D130A, D130N and R131L were non functioning. Tyrosine 131 was particularly sensitive to mutation as both Y131F and Y131S mutants were poorly expressed and were chemotactically inactive. Together, this data suggest that the acidic / basic / polar nature of the DRY motif is a prerequisite for CCR3 function. PMID- 11920573 TI - A human TAPBP (TAPASIN)-related gene, TAPBP-R. AB - TAPASIN, a V-C1 (variable-constant) immunoglobulin superfamily (IgSF) molecule that links MHC class I molecules to the transporter associated with antigen processing (TAP) in the endoplasmic reticulum (ER) is encoded by the TAPBP gene, located near to the MHC at 6p21.3. A related gene was identified at chromosome position 12p13.3 between the CD27 and VAMP1 genes near a group of MHC-paralogous loci. The gene, which we have called TAPBP-R (R for related), also encodes a member of the IgSF, TAPASIN-R. Its putative product contains similar structural motifs to TAPASIN, with some marked differences, especially in the V domain, transmembrane and cytoplasmic regions. By using the mouse ortholog to screen tissue, we revealed that the TAPBP-R gene was broadly expressed. Sub-cellular localization showed that the bulk of TAPASIN-R is located within the ER but biotinylation experiments were consistent with some expression at thecell surface. TAPASIN-R lacks an obvious ER retention signal. The function of TAPASIN R will be of interest in regards to the evolution of the immune system as well as antigen processing. PMID- 11920574 TI - T lymphocyte expression of thrombospondin-1 and adhesion to extracellular matrix components. AB - The mechanisms controlling the formation of pseudopodia and other active cell edges in T lymphocytes are not understood. We show here that T lymphocytes express thrombospondin-1 (TSP-1). TSP-1 in T lymphocytes has a high turnover as shown by the fact that brefeldin and monensin rapidly increase while cycloheximide tend to decrease the cellular TSP-1 content. T cell TSP-1 is preferentially stored intracellularly and shows variable cell surface expression. T lymphocyte adhesion to fibronectin and collagen type IV induces TSP-1 expression on the cell surface via a brefeldin sensitive mechanism. A monoclonal antibody to TSP-1 inhibits the flattening and pseudopodia formation of the adherent T cells. Furthermore, the same antibody to TSP-1 also exerts an inhibitory effect on T cell migration in the absence of exogenous TSP-1. These results indicate that endogenous TSP-1 is part of an adhesion-dependent mechanism controlling cytoplasmic spreading and migration in T lymphocytes. PMID- 11920575 TI - Functional restoration of human immunodeficiency virus and Epstein-Barr virus specific CD8(+) T cells during highly active antiretroviral therapy is associated with an increase in CD4(+) T cells. AB - To investigate the effect of highly active antiretroviral therapy (HAART) on HIV- and Epstein-Barr virus (EBV)-specific CD8(+) T cells, total number and function of these cells was determined in 16 HIV-infected individuals using tetrameric HLA peptide complexes and IFN-gamma ELISPOT assays after peptide stimulation, respectively. HAART induced a significant decrease in HIV-specific tetramer(+) T cells, whereas EBV-specific tetramer(+) T cells did not change. In addition, individuals who temporarily failed on therapy showed a temporary increase in the number of HIV-specific T cells, suggesting that differences in the pool size of antigen-specific T cells was determined by the presence of antigen. Interestingly, there was an increase in the ratio of IFN-gamma-producing T cells per total number of both HIV- and EBV-specific T cells in the majority of individuals, suggesting that the function of virus-specific T cells is improved in individuals successfully treated with HAART. Despite this relative functional improvement of EBV-specific T cells, no significant changes were observed in EBV load. In four subjects who temporarily failed on HAART, the percentage of IFN gamma-producing T cells, both for HIV and EBV, paralleled CD4(+) T cell kinetics, suggesting that function seems to be related to differences in CD4(+) T cell numbers. Overall, these data indicate that HAART improves the antigen responsiveness of both HIV- and EBV-specific T cells, which is associated with an increase in CD4(+) T cells. PMID- 11920576 TI - Regulation of activation-induced receptor activator of NF-kappaB ligand (RANKL) expression in T cells. AB - Receptor activator of NF-kappaB ligand (RANKL) is a type II membrane protein of the TNF family and plays a critical role in the regulation of osteoclastogenesis. RANKL expressed on osteoblastic stromal cells has been shown to support osteoclast differentiation originated from hematopoietic precursors. Interestingly, RANKL is also expressed on cells of the immune system including T cells and dendritic cells. We have shown that anti-CD3 could induce RANKL expression in T cell hybridoma A1.1 cells and splenic T cells. RANKL expressed on T cells could effectively induce osteoclast formation from the whole population of murine splenocytes. Furthermore, we have found that the induction of RANKL expression is solely dependent on TCR activation-induced Ca2+ mobilization since its expression can be blocked by cyclosporine A and TMB-8, a Ca2+ mobilization inhibitor. Additionally, treatment of A1.1 cells with ionomycin alone also strongly induces RANKL expression, while phorbol myristate acetate by itself does not. Moreover, although inhibition of c-myc has significant effects on anti-CD3 induced Fas ligand (FasL) expression, we have found that the anti-CD3-induced RANKL expression is independent of c-myc. Surprisingly, in contrast to its inhibitory effect on FasL expression, TGF-beta dramatically increased the expression of anti-CD3-induced RANKL expression. In addition to its potential role in immune responses, RANKL expressed on activated T lymphocytes may provide a mechanism for the communication between the immune and skeletal systems during immune responses and disease states such as rheumatoid arthritis. PMID- 11920577 TI - Dendritic cells pulsed with exogenous hepatitis B surface antigen particles efficiently present epitopes to MHC class I-restricted cytotoxic T cells. AB - L(d)- and K(b)-binding epitopes processed by murine dendritic cells (DC) pulsed with exogenous, particulate hepatitis B surface antigen (HBsAg) are presented to cytotoxic T lymphocytes (CTL). The specific and dose-dependent induction of IFN gamma release and cytotoxicity in CTL by metabolically active DC did not depend on antigenic peptides contaminating the particles, was cytochalasin D resistant, independent of the maturation state of DC, and blocked by primaquine, amiloride and NH(4)Cl (indicating involvement of acid proteolysis). The specific immunostimulatory phenotype of pulsed DC was maintained for about 3 h after the end of the pulse but rapidly decayed thereafter. Processing of L(d)- and K(b) binding epitopes from exogenous HBsAg particles by pulsed DC for presentation was TAP independent. Surface-associated 'empty' (presentation-deficient) 64(+) L(d) molecules (defined by the mAb 64-3-7), but not trimeric (presentation-competent) 30(+) L(d) molecules (defined by the mAb 30-5-7) had to be available during the pulse of DC with exogenous HBsAg particles to generate 30(+) L(d)molecules that present the antigenic S(28-39) peptide. Exogenous beta2-microglobulin present during the pulse of DC with HBsAg particles facilitated presentation of L(d)- and K(b)-restricted epitopes. DC generated from bone marrow progenitors in vitro, as well as splenic and liver DC (generated in vivo) presented epitopes to specific CTL. HBsAg particles thus efficiently enter an alternative processing pathway in DC that leads to presentation of epitopes to MHC class I-restricted CTL. PMID- 11920578 TI - Mesenteric lymph nodes are critical for the induction of high-dose oral tolerance in the absence of Peyer's patches. AB - We have previously demonstrated the loss of oral tolerance (OT) in lymphotoxinalpha-/- (LTalpha-/-) and TNFalpha / lymphotoxinalpha deficient (TNFalpha / LTalpha-/-) mice which have defective Peyer's patches (PP) and lymph node (LN) development. We have now studied OT in BALB / c mice with differential defects of the gut-associated lymphoid tissue (GALT) caused by inhibition of LTbetaR signaling during fetal development. Treatment of pregnant mice with LTbetaR-IgG (LTbetaRIgG) and TNFR I55-IgG (TNFR55IgG) abrogates the formation of PP (LTbetaRIgG) or of PP and mesenteric LN (MLN) (LTbetaRIgG / TNFRIgG) without genetically deleting the respective cytokine pathways. OT was readily induced in mice without PP but retaining MLN (PP null / LN +). In contrast, OTcould not be induced in mice lacking both MLN and PP (PP null / MLN null) as shown by the inability of these mice to suppress IFN-gamma secretion or DTH reactions. We next assessed OT in 129 x B6 LTalpha-/- mice with and without MLN. Timed treatment of pregnant LTalpha-/- mice with an agonist anti-LTbetaR mAb induces formation of MLN but not of PP in LTalpha-/- mice. LN + LTalpha-/- mice developed OT while LN LTalpha-/- mice were resistant to OT induction. Taken collectively, the data show that in the presence of MLN PP are not required for OT induction and that the presence of MLN is sufficient for OT induction in the LTalpha-/- model. PMID- 11920579 TI - A role for antibody and Fc receptor in the clearance of Brugia malayi microfilariae. AB - The roles of antibodies (Ab) and Fc receptors (FcR) in clearance of Brugia malayi microfilariae (Mf) in vivo have not been previously elucidated. Different background strainsof mice clear Mf at different rates and we established that there were no major differences in Ab production between three particular strains. However, genetic immunodeficiencies in B cell and Ab production on each of these background strains significantly enhanced Mf survival. B cell-deficient microMT (C57BL/6) mice injected with B. malayi Mf i.v. retained significant numbers of live Mf in comparison to wild-type mice. Treatment of microMT mice with hyperimmune whole sera in the first week of infection significantly reduced the number of Mf surviving in the cardiac blood at 135 days p.i. Mf survival in FcRgamma-deficient (Sv129xC57BL/6) mice was also dramatically increased in the cardiac blood compared to wild-type mice, indicating that Ab itself is involved in Mf clearance most likely via a mechanism involving ADCC. These data indicate that Ab plays an important role in vivo in Mf killing in mice and this role is mediated via the FcR. PMID- 11920580 TI - SWAP-70-deficient mast cells are impaired in development and IgE-mediated degranulation. AB - Cross-linking of the high-affinity IgE receptor (FcepsilonRI) on mast cell activates signaling pathways that trigger degranulation and the release of multiple pro-inflammatory mediators. Mature,immature and precursor mast cells are degranulation competent. We show here that the signaling protein SWAP-70 has a function in mast cell biology. While not found in many cell types, we find that apart from B cells, mast cells also express SWAP-70. In activated B cells, SWAP 70 shuttles between cytoplasm and nucleus, but in mast cells it is confined to the cytoplasm. SWAP-70(ko/ko) (double knockout) mice have reduced numbers of mature mast cells, and these are degranulation competent. However, although immature mast cells from SWAP-70(ko/ko) mice respond normally to SCF and IL-3 and have functional granules, their FcepsilonRI-mediated degranulation is inhibited. Thus, in mast cells SWAP-70 plays a role both in establishing the initial competence to degranulate and to develop into mature mast cells. PMID- 11920581 TI - Cancer chemoresistance: the relationship between p53 and multidrug transporters. AB - Extensive studies indicate that both p53 and multidrug transporters play important roles in chemoresistance. Since the initial reports a decade ago demonstrating a transcriptional dependence of the ABCB1 gene (MDR) promoter by p53, much data have been accumulated. However, despite being the subject of intense study, this p53-MDR relationship remains unclear in human cancers. The data are confounded by variable and contrasting results when considering the in vitro regulation and attempting to draw parallels in tissue specimens. The original model suggested that wild-type p53 downregulates the ABCB1 promoter, whereas mutant p53 increases expression of ABCB1. This review summarizes the data for and against this hypothesis. What emerges from these studies is a complex picture, where data have been obtained in support of this hypothesis, but there are also many circumstances where it is not supported. Taken together, these data suggest that the relationship between p53 and multidrug transporters is conditional. It is dependent on cellular environment, the drug used, and the nature of the p53 mutation. PMID- 11920582 TI - In situ repair of cyclobutane pyrimidine dimers in skin and melanocytic nevi of cutaneous melanoma patients. AB - The development of cutaneous malignant melanoma (CMM) and its precursor lesions, melanocytic nevi, has been linked to sun exposure. Cyclobutane pyrimidine dimers (CPDs) are the majority of DNA lesions induced by sun exposure. In our study, we investigated if CMM patients have impaired ability to repair CPDs in skin as well as in melanocytic nevi. The repair kinetics were followed up to 3 weeks after exposure to 40 mJ/cm(2) of solar simulating radiation. Altogether 12 CMM patients and 10 healthy controls were included in our study. Buttock skin biopsies were taken at 0 hr, 48 hr and 3 weeks after UV exposure, whereas melanocytic nevi and surrounding skin biopsies were taken only at 0 hr and 3 weeks. The CPD levels were measured by a (32)P-postlabeling method. The results showed that the repair rate of CPDs in neither the skin nor the nevi was significantly different between the CMM patients and the control group. For both groups, the repair rate of TT = C was faster than that for TT = T. The important finding is that about 10% of the initial TT = T damage remained unrepaired after 3 weeks, and was detectable in normal epidermis as well as in nevi of all subjects. We also found that the amount of TT = C and TT = T at 0 hr in nevi was significantly lower than that in surrounding skin (Wilcoxon rank sum test, p < 0.05). PMID- 11920583 TI - Correlation between vascular endothelial growth factor-C expression and invasion phenotype in cervical carcinomas. AB - The correlation between vascular endothelial growth factor (VEGF)-C gene expression and in vitro invasive activity and matrix metalloproteinase (MMP)-2 or 9 gene expression and proteolytic activity in 11 cervical carcinoma cell lines, was investigated. Immunohistochemical expression of VEGF-C in 52 cervical carcinoma tissues was also correlated with tumor aggressiveness with respect to clinicopathologic features, tumor vascularity, MMP-2 expression and patient outcome. Expression of VEGF-C mRNA differed remarkably among the cell lines and there was a statistical correlation between VEGF-C gene expression and the number of invaded tumor cells (p = 0.0009) and MMP-2 gene expression and activity (p < 0.05). Anti-VEGF-C antibody inhibited the invasive and proteolytic activity of tumor cells in a concentration-dependent manner. VEGF-C or MMP-2 expression in clinical tissue samples was well correlated with depth of myometrial invasion, endometrial invasion, pelvic lymphnode metastasis and tumor vascularity (p < 0.05) and there was a close relation between VEGF-C and MMP-2 expression (p < 0.0001) in cervical carcinomas. Overall survival rates for 14 patients with strong VEGF-C staining tumors were lower than those for 38 patients with weak VEGF-C staining tumors (p = 0.0132) and VEGF-C tissue status emerged as an independent prognostic parameter (p = 0.0232). These results suggest that VEGF-C expression is closely related to invasion phenotype and affects the patient's survival in cervical carcinomas. PMID- 11920584 TI - Troglitazione affects survival of human osteosarcoma cells. AB - Activation of PPAR gamma, a transcription factor member of the family of peroxisome proliferator-activated receptors, induces apoptosis in several normal and tumor cell lines. In our study, we investigated whether treatment with troglitazone (TRO), a known PPAR gamma agonist, induced apoptosis in the human osteosarcoma (OS) cell lines G292, MG63, SAOS and U2OS that express PPAR gamma. In our experiments, TRO never induced apoptosis of OS cells; on the contrary, TRO increased cell number, based on MTT proliferation assay. Remarkably, the TRO induced cell number increase depended on a decrease of apoptosis that naturally occurred in the culture and was not due to an increased cell proliferation rate. TRO also prevented staurosporin-induced apoptosis. The TRO-mediated survival effect correlated with the activation of Akt, a well-known mediator of survival stimuli. Our work describes a new function for TRO and indicates that the Akt survival pathway may be a mediator of TRO-induced increase of survival. PMID- 11920585 TI - CD70/CD27 ligand, a member of the TNF family, is expressed in human brain tumors. AB - CD70 (CD27 ligand) has been implicated in proapoptotic signals mediated by its receptor CD27 in lymphocytes as well as in proliferative effects induced by reverse signaling in CD70-positive hematopoetic tumor cells. We were able to show that CD70 is expressed at the mRNA and protein level in human meningioma cells, glioma cells from solid human gliomas as well as glioma cell lines and 1 ependynoma. The intensity of CD70 expression varies considerably between different samples from 1 tumor type. In the U343 glioma cell line, CD70 was preferentially localized in the cell processes. Thus, our studies identify CD70 as a new marker molecule in brain tumors that are of nonhematopoetic origin. PMID- 11920586 TI - Evidence of high incidence of EGFRvIII expression and coexpression with EGFR in human invasive breast cancer by laser capture microdissection and immunohistochemical analysis. AB - EGFRvIII was first reported in human glioblastomas. Subsequent reports indicated EGFRvIII protein to be frequently detected in several other human cancers, but not in normal tissues. Our previous studies suggested that EGFRvIII could induce a transformation from ligand-dependent non-tumorigenic cell line to ligand independent malignant phenotype cells in vitro and in vivo. Transfection of EGFRvIII in MCF-7 cell line resulted in a 3-fold increase in colony formation and significantly enhanced tumorigenicity in nude mice (p < 0.001). EGFRvIII could also induce ErbB-2 phosphorylation. The existence and significance of EGFRvIII transcript in human breast cancer, however, was not reported. In our study, we detected the presence of EGFRvIII mRNA and revealed a high incidence (67.8%) of EGFRvIII transcript in human primary invasive breast cancer by utilizing laser capture microdissection (LCM)/RT-PCR to capture pure breast cancer cells. In addition, 57.1% of the infiltrating breast carcinomas expressed both EGFRwt and EGFRvIII mRNA in the same tumor. There is no detectable EGFRvIII mRNA in normal breast tissue. Evaluation of the EGFRwt and EGFRvIII protein levels in the same sample sets by immunohistochemical analysis further confirmed the LCM/RT-PCR finding. Our study provides first direct evidence of high incidence of coexpression of EGFRvIII and EGFRwt in human invasive breast cancer tissue. The unique characteristics and high prevalence of EGFRvIII in invasive human breast cancer as well as negative expression in normal breast may suggest its important role in breast carcinogenesis and make it an ideally potential target for treatment of breast cancer without interrupting normal EGFR signaling. PMID- 11920587 TI - Clinical impact and functional aspects of tenascin-C expression during glioma progression. AB - The extracellular matrix protein tenascin-C is expressed in processes like embryogenesis and wound healing and in neoplasia. Tenascin-C expression in gliomas has been described previously; however, the relation to clinical data remains inconsistent. Generally, analysis of tenascin-C function is difficult due to different alternatively spliced isoforms. Our studies focus on changes in tenascin-C expression in human gliomas, correlating these changes with tumor progression and elucidating the functional role of the glioma cell-specific tenascin-C isoform pool. Eighty-six glioma tissues of different World Health Organization (WHO) grades were analyzed immunohistochemically for tenascin-C expression. The influence of the specific tenascin-C isoforms produced by glioblastoma cells on proliferation and migration was examined in vitro using blocking antibodies recognizing all isoforms. In general, tenascin-C expression increased with tumor malignancy. Perivascular staining of tenascin-C around tumor supplying blood vessels was observed in all glioblastoma tissues, whereas in WHO II and III gliomas, perivascular tenascin-C staining appeared less frequently. The appearance of perivascular tenascin-C correlated significantly with a shorter disease-free time. Analysis of proliferation and migration in the presence of blocking antibodies revealed an inhibition of proliferation by around 30% in all 3 glioblastoma cell cultures, as well as a decrease in migration of 30.6-46.7%. Thus we conclude that the endogenous pool of tenascin-C isoforms in gliomas supports both tumor cell proliferation and tumor cell migration. In addition, our data on the perivascular staining of tenascin-C in WHO II and III gliomas and its correlation with a shorter disease-free time suggest that tenascin-C may be a new and potent prognostic marker for an earlier tumor recurrence. PMID- 11920588 TI - Transcriptional inactivation of TP73 expression in oligodendroglial tumors. AB - The TP73 gene, located on chromosome 1p36.3, encodes a product that shares significant structural homology with the tumor suppressor TP53. The aim of this study was to investigate whether TP73 is involved in the development of oligodendroglial tumors, which frequently carry deletions involving 1p36.3. Semi quantitative reverse transcription-polymerase chain reaction was used to determine TP73 transcript levels. Ten of 24 (42%) tumors showed negligible to more than 5-fold reduction in TP73 expression when compared to normal brain level. To identify potential mechanisms that may modulate TP73 transcription in oligodendroglial tumors, we performed mutation analysis on the TP73 gene. No somatic mutations were however detected in the gene sequence. We then evaluated the possible involvement of epigenetic change in TP73 expression. Bisulfite genomic sequencing detected aberrant hypermethylation at the 5' region upstream and including the first exon of the TP73 gene in 17 of 44 (39%) oligodendroglial tumors, whereas normal brain tissues showed no methylation in the same region examined. Moreover, 6 of 10 (60%) tumors with negligible or decreased levels of TP73 transcripts were methylation-positive. In conclusion, our results showed that inactivation of TP73 occurs at the transcriptional level and is associated with promotor hypermethylation. Loss of or reduced TP73 transcript expression may contribute to the tumorigenesis of oligodendroglial tumors. PMID- 11920589 TI - Peptide-specific CD8+ T-cell evolution in vivo: response to peptide vaccination with Melan-A/MART-1. AB - Monitoring of CD8+ T-cell responses in cancer patients during peptide vaccination is essential to provide useful surrogate markers and to demonstrate vaccine efficacy. We have longitudinally followed CD8+ T-cell responses in 3 melanoma patients who were immunized with peptides derived from Melan-A/MART-1. Recombinant HLA-A2 tetramers loaded with the naturally presented Melan-A/MART-1 nonamer peptide (AAGIGILTV) and the Melan-A/MART-1 analog (ELAGIGILTV) were used in combination with phenotypical analysis for different T-cell subsets including naive T cells, effector T cells, "true memory" T cells and "memory effector" T cells, based on CD45RA/RO and CCR7-expression. At least in a single patient, T cells binding to the AAGIGILTV epitope were detected in naive, precursor (CD45RA+/CCR7+) CD8+ T cells, and CD8+ T cells binding to the analog ELAGIGILTV peptide were identified in the terminally differentiated (CD45RA+/CCR7-) T-cell subset. Molecular and functional analysis of tetramer-binding T cells revealed that the T-cell receptor (TCR) repertoire was oligo/polyclonal in AAGIGILTV reactive T cells, but different and restricted to a few TCR clonotypes in ELAGIGILTV-reactive T cells prior to vaccination. The TCR repertoire reactive with Melan-A/MART-1 peptide epitopes was broadened during vaccination and exhibited a different profile of cytokine release after specific stimulation: tetramer-binding T cells from 2/3 patients secreted granulocyte/macrophage colony stimulating factor (GM-CSF) and interferon-gamma but not interleukin-2 (IL-2) in response to Melan-A/MART-1 peptides. In the third patient, tetramer-binding T cells secreted IL-2 exclusively. Our results show that T-cell responses to peptide vaccination consist of different T-cell subsets associated with different effector functions. Complementary analysis for TCR CDR3 and cytokine profiles may be useful to define the most effective CD8+ T-cell population induced by peptide vaccination. PMID- 11920590 TI - Regulation of CD1d expression by murine tumor cells: escape from immunosurveillance or alternate target molecules? AB - alpha beta+ TCR T cells recognize peptide fragments displayed by MHC-class I or class II molecules. Recently, additional mechanisms of antigen recognition by T cells have been identified, including CD1-mediated presentation of nonpeptide antigens. Only a limited number of CD1 antigens is retained in the mouse, i.e., the group II CD1 antigens, which are split into CD1D1 and CD1d2. Several T cell subsets have been shown to interact with murine CD1 antigens, including NK cells or "natural T cells" with the invariant V alpha 14 J alpha 281 TCR chain. Even if TAP defects may prevent classical endogenous antigen presentation in tumor cell lines, antigen presentation via CD1 is still functional. Therefore, CD1-mediated recognition of transformed cells by NK cells or "natural T cells" may represent an alternative way for immune surveillance. CD1 cell surface expression in murine tumor cell lines of different histology, including the B cell lymphoma A20, macrophage cell lines J774 and P388D1, mastocytoma P815, thymoma EL-4, melanoma B16, colon adenocarcinoma MC-38 and renal carcinoma Renca is regulated by Th1- (IFN-gamma), Th2- (IL-4, IL-10 and vIL-10) or GM-CSF (Th1/Th2) cytokines, depending on the tumor histology. In order to distinguish between CD1D1 and CD1d2 molecules, we examined differential expression of these CD1 isoforms by ratio RT PCR: A20, EL-4, P815 and MC-38 cells exclusively express CD1D1 transcripts but not CD1D2 mRNA independent of cytokine treatment. Decreased CD1d expression leads to reduced immune recognition of CD1d+ tumor cells by freshly isolated NK1.1(+) effector cells as defined by cytolysis and IFN-gamma release. Thus, modulation of CD1 expression on tumor cells by cytokines may be advantageous to drive cellular anti-tumor antigen directed immune responses directed against TAP-independent, non-classical MHC restricting molecules. PMID- 11920591 TI - Novel monoclonal antibody specific for the de2-7 epidermal growth factor receptor (EGFR) that also recognizes the EGFR expressed in cells containing amplification of the EGFR gene. AB - In some respects, the EGFR appears to be an attractive target for tumor-targeted antibody therapy: it is overexpressed in many types of epithelial tumor and inhibition of signaling often induces an anti-tumor effect. The use of EGFR specific antibodies, however, may be limited by uptake in organs that have high endogenous levels of the wild type EGFR such as the liver. The de2-7 EGFR (or EGFRvIII) is a naturally occurring extracellular truncation of the EGFR found in a number of tumor types including glioma, breast, lung and prostate. Antibodies directed to this tumor specific variant of the EGFR provide an alternative targeting strategy, although the lower proportion of tumors that express the de2 7 EGFR restricts this approach. We describe a novel monoclonal antibody (MAb 806) that potentially overcomes the difficulties associated with targeting the EGFR expressed on the surface of tumor cells. MAb 806 bound to de2-7 EGFR transfected U87MG glioma cells (U87MG.Delta 2-7) with high affinity (approximately 1 x 10(9) M(-1)), but did not bind parental cells that express the wild type EGFR. Consistent with this observation, MAb 806 was unable to bind a soluble version of the wild type EGFR containing the extracellular domain. In contrast, immobilization of this extracellular domain to ELISA plates induced saturating and dose response binding of MAb 806, suggesting that MAb 806 can bind the wild type EGFR under certain conditions. MAb 806 also bound to the surface of A431 cells, which due to an amplification of the EGFR gene express large amounts of the EGFR. Interestingly, MAb 806 only recognized 10% of the total EGFR molecules expressed by A431 cells and the binding affinity was lower than that determined for the de2-7 EGFR. MAb 806 specifically targeted U87MG.Delta 2-7 and A431 xenografts grown in nude mice with peak levels in U87MG.Delta 2-7 xenografts detected 8 h after injection. No specific targeting of parental U87MG xenografts was observed. Following binding to U87MG.Delta 2-7 cells, MAb 806 was rapidly internalized by macropinocytosis and subsequently transported to lysosomes, a process that probably contributes to the early targeting peak observed in the xenografts. Thus, MAb 806 can be used to target tumor cells containing amplification of the EGFR gene or de2-7 EGFR but does not bind to the wild type EGFR when expressed on the cell surface. PMID- 11920592 TI - Identification of known and novel immunogenic T-cell epitopes from tumor antigens recognized by peripheral blood T cells from patients responding to IL-2-based treatment. AB - In previous studies CD8+ T cells specific for melanocyte antigens have been frequently found in melanoma patients responding to interleukin-2 (IL-2)-based therapies. In our study we analyzed the suitability of using circulating T cells from melanoma patients with clinical response after IL-2-based therapy to identify novel T-cell epitopes from defined tumor antigens. Using unstimulated peripheral blood mononuclear cells and the interferon-gamma (IFN-gamma) ELISPOT assay, we studied CD8(+) T-cell responses against 5 peptides from the tumor antigen tyrosinase (Tyr) selected by epitope prediction using an HLA-A1-binding computer algorithm. T cells specifically secreting IFN-gamma in response to 3 of these 5 peptides, namely, Tyr (454-463), Tyr (146-156) and Tyr (243-251), could be detected in 4 of 4 HLA-A1-positive patients with clinical response. In contrast, no T-cell responses against these peptides were seen in 6 HLA-A1 positive melanoma patients with progressive disease and in 8 healthy subjects. We could generate specific cytotoxic T lymphocytes (CTL) against Tyr (454-463) using peptide-pulsed autologous dendritic cells as antigen-presenting cells. The induced CTLs efficiently killed melanoma cells that express HLA-A1 and tyrosinase. The peptides Tyr (146-156) and Tyr (243-251) had recently been identified as CTL epitopes by other groups. Further ex vivo characterization of the T cells reactive against the novel epitope Tyr (454-463) in 1 patient by multicolor flow cytometry showed specific CD3+/CD8+/IFN-gamma+ T cells with frequencies of up to 0.41% of the CD3+/CD8+ T-cell population. Most of this T cell population also expressed granzyme B. Our data confirm that in patients with tumor regressions induced by immunotherapy or chemoimmunotherapy circulating T cells reactive with tyrosinase epitopes can frequently be detected. Peripheral blood T cells from such patients are a valuable source for screening peptides selected by epitope prediction This strategy facilitates the rapid identification of immunogenic T-cell epitopes that are probable targets of immune-mediated tumor rejection. PMID- 11920593 TI - Overexpression of cyclinD1 predicts for poor prognosis in estrogen receptor negative breast cancer patients. AB - CyclinD1 plays a critical role in regulating cell cycle progression. CyclinD1 mRNA and protein are overexpressed in approximately 50% of primary breast cancer cases. However, its clinical significance as a predictive factor remains unclear. One hundred and seventy-three female patients diagnosed with invasive ductal carcinoma who had undergone a mastectomy (161 patients) or breast-conserving surgery (12 patients) were followed up for 6-119 months (median 86 months) postoperatively. Immunoreactivity for monoclonal anti-cyclinD1 antibody (clone DCS-6) with paraffin-embedded carcinoma tissues was investigated using a labeled streptavidin-biotin method. Overexpression of cyclinD1 was found in 42% (73 of 173), and strongly correlated with estrogen receptor (ER) expression (p < 0.000001). Univariate analysis revealed no association between overexpression of cyclinD1 and overall survival or relapse-free survival in all patient groups. However, in the ER-negative subgroup (n = 75), overexpression of cyclinD1 was significantly correlated with shorter overall survival (p = 0.018) and relapse free survival (p = 0.014) as well as the lymph node status and tumor size. In contrast, there were no significant associations between overexpression of cyclinD1 and clinical outcome in the ER-positive subgroup. According to Cox's multivariate analysis in the ER-negative subgroup, overexpression of cyclinD1 had the most significant effect on overall survival (p = 0.02) and relapse-free survival (p = 0.0058), followed by nodal status and histologic grade. These findings suggest that overexpression of cyclinD1 is an independent prognostic indicator in ER-negative breast cancer patients. PMID- 11920594 TI - Identification and characterization of genes involved in the carcinogenesis of human squamous cell cervical carcinoma. AB - We utilized RT-PCR differential display and cDNA microarrays to identify cellular genes involved in the multi-step carcinogenesis of squamous cell cervical carcinoma. Thirty-eight cervical cancer patients in various stages of the disease and 5 non-cervical cancer patients were studied. Twenty-five cDNA clones were identified and these were subsequently demonstrated to be consistently over expressed in squamous cell cervical carcinoma biopsies of various FIGO stages. To further evaluate the possible role that these genes may play in the progression of disease, we performed Northern blot analysis and RNA-RNA in situ hybridization studies using cervical cancer biopsies of various FIGO stages. Of particular interest are the 2 clones G32C4B and G30CC that have been identified to be the NADH dehydrogenase 4 gene and the gene that encodes ribosomal protein S12 respectively when compared to sequences available in the GenBank database. Increased expression of these 2 genes were detected in the matched normal tissues collected together with the late FIGO stages of cervical cancer biopsies. In comparison, upregulation of these 2 genes was not detected in cervical squamous epithelium collected from patients admitted for surgery for non-malignant conditions, suggesting that expression of these 2 genes may have altered in the adjacent histopathologically "normal" cervical squamous epithelial tissue from cervical cancer patients. The ribosomal protein S12 and the NADH dehydrogenase 4 genes may therefore be potentially useful as early pre-transformation diagnostic markers for human cervical cancer. PMID- 11920595 TI - Expression of cyclooxygenase-2 in the subserosal layer correlates with postsurgical prognosis of pathological tumor stage 2 carcinoma of the gallbladder. AB - Postsurgical recurrence at distant sites frequently occurs in pathological tumor stage 2 (pT(2)) carcinoma of the gallbladder even though the carcinoma is limited to the gallbladder wall. Little is known, however, about the molecular events leading to its development and progression. A large body of evidence suggests that cyclooxygenase-2 (COX-2) is up-regulated in carcinoma tissues and plays roles in promoting cell-proliferation, growth and metastasis of carcinoma cells. In the present study, immunohistochemistry was performed to determine the expression levels of COX-2 in the subserosal layer of 33 cases of pT(2) gallbladder carcinoma in which curative resections had been performed and to determine the correlations of the expression levels of COX-2 with mode of recurrence and postsurgical survival. Immunostaining of COX-2 in the epithelia was recognized in more than 80% of normal epithelia, noncancerous pathological lesions of the gallbladder except for intestinal metaplasia and pT(1-4) carcinoma specimens. Intense staining was observed in large percentages of hyperplastic lesions (65%), pT(2) carcinoma specimens (76%) and pT(3) and pT(4) carcinoma specimens (64%) compared to the percentages of normal epithelia and other pathological lesions (0-25%). Intense staining was also observed in the adjacent stroma in pT(2) carcinoma specimens (33%) and in those in pT(3) and pT(4) carcinoma specimens (43%) but only in small percentages of the stroma adjacent to normal epithelia and pathological lesions (0-8%). In situ hybridization confirmed the existence of COX-2 mRNA in both the cancerous epithelia and adjacent stroma of pT(2)-pT(4) carcinomas. In 33 cases of pT(2) carcinoma, distant recurrence, i.e., liver metastasis, was seen in 3 of 9 cases of pT(2) carcinoma (33%, P<0.05) with intense stromal staining in the subserosal layer and in 1 of 24 cases (4%) without intense staining, whereas no significant correlation was found between parameters of pathological malignancies (histological grade, lymphatic permeation, venous permeation and lymph node metastasis) and the intensity of stromal staining in the subserosal layer. The postsurgical survival outcome was significantly poorer in the former than in the latter (p = 0.010). In pT(2) gallbladder carcinoma, upregulation of COX-2 in the stroma adjacent to the cancerous epithelia in the subserosal layer correlates with the aggressiveness of the disease, such as the tendency to form distant recurrences. This phenotype may serve as a unique biological feature associated with the malignant behavior of pT(2) gallbladder carcinoma. PMID- 11920596 TI - Persistent human papillomavirus infection and smoking increase risk of failure of treatment of cervical intraepithelial neoplasia (CIN). AB - Women with abnormal smears have an increased risk of developing cervical cancer. During the 8 years following conservative treatment of cervical intraepithelial neoplasia (CIN), their risk of invasive cervical cancer is about 5 times greater than that of the general population. Human papillomavirus (HPV) has been associated with the natural history of both CIN and cervical cancer. To date, there have been no published reports on the predictive value of HPV testing in the treatment outcome of CIN. A prospective, multi-center, cohort study was conducted on women in the Northwest of England who were attending for treatment of CIN. They were asked to complete a questionnaire, which included a detailed smoking history. Pre- and post-treatment HPV testing was performed on cervical biopsies and cervical swab, being taken with the first follow-up smear at 6 months. A nested case-control analysis was performed, cases being defined as women who developed CIN within the 2 years of treatment and controls being sampled from those who did not experience treatment failure within 2 years. Multiple conditional logistic regression is used to study the factors associated with treatment failure of CIN. The cohort included 958 women of whom 77 (8%) experienced treatment failure (cases). Two controls were matched to each case (154). Smoking status was significantly associated with CIN treatment failure(p= 0.0013). Current smokers had a 3-fold increased risk of treatment failure of CIN as compared to non-smokers (95% CI 1.65 to 5.91). Five hundred twenty-five women underwent HPV sampling following treatment, of whom 47 (8.9%) developed further CIN. Post-treatment positive HPV testing was found to be strongly associated with treatment failure of CIN (OR 23.3; 95% CI 3.15-172.1). In 11/45 cases with negative smear at first follow-up, the HPV test was positive. The combination of both HPV and cytology in the first follow-up visit predicted treatment failure in 72% of the cases. Cigarette smoking is a factor, which, independently of HPV infection, influences the treatment outcome of CIN. Smokers and those who are HPV positive during follow-up appear to require longer, more intensive follow-up. HPV testing requires careful consideration as part of routine follow-up protocol following treatment of CIN. PMID- 11920597 TI - Oral cancer in southern India: the influence of smoking, drinking, paan-chewing and oral hygiene. AB - Between 1996 and 1999 we carried out a case-control study in 3 areas in Southern India (Bangalore, Madras and Trivandrum) including 591 incident cases of cancer of the oral cavity (282 women) and 582 hospital controls (290 women), frequency matched with cases by age and gender. Odds ratios (ORs) and 95% confidence intervals (CIs) were obtained from unconditional multiple logistic regressions and adjusted for age, gender, center, education, chewing habit and (men only) smoking and drinking habits. Low educational attainment, occupation as a farmer or manual worker and various indicators of poor oral hygiene were associated with significantly increased risk. An OR of 2.5 (95% CI 1.4-4.4) was found in men for smoking > or = 20 bidi or equivalents versus 0/day. The OR for alcohol drinking was 2.2 (95% CI 1.4-3.3). The OR for paan chewing was more elevated among women (OR 42; 95% CI 24-76) than among men (OR 5.1; 95% CI 3.4-7.8). A similar OR was found among chewers of paan with (OR 6.1 in men and 46 in women) and without tobacco (OR 4.2 in men and 16.4 in women). Among men, 35% of oral cancer is attributable to the combination of smoking and alcohol drinking and 49% to pan tobacco chewing. Among women, chewing and poor oral hygiene explained 95% of oral cancer. PMID- 11920598 TI - Risk of gastric cancer among smokers infected with Helicobacter pylori. AB - Infection with the gastric bacterium Helicobacter pylori (in particular infection with CagA-positive strains) and smoking have been identified as risk factors for the development of gastric cancer. Both risk factors are typically acquired early in life and prevail over decades if not for life. We assessed the individual and joint impact of both risk factors on gastric cancer risk in a population-based case-control study from Germany including 71 patients with histologically verified gastric cancer and 363 patients with colorectal cancer who served as controls. Information on smoking and potential confounding factors was collected by standardized interviews. H. pylori infection was measured serologically by immunoglobulin G antibody titers against H. pylori. In addition, antibodies against the CagA antigen were determined by Western blot. Twenty-seven percent of cases compared with 15% of controls were smokers, and 43% of cases compared with 23% of controls were infected with CagA-positive H. pylori strains. After control for potential confounders, the relative risk of gastric cancer was 2.6 (95% CI 1.2-5.7) for nonsmoking subjects with CagA-positive H. pylori infections and 7.2 (95% CI 2.2-23.6) for smoking subjects with CagA-positive H. pylori infections compared with subjects without these risk factors. The corresponding relative risks for noncardia gastric cancer were 6.1 ( 95% CI 2.3-16.5) and 16.6 (95% CI 4.3-64.2). We conclude that smoking subjects with CagA-positive H. pylori infections have a strongly increased risk of gastric cancer and may be an important group for targeting efforts of prevention and early detection. PMID- 11920599 TI - Prediction of chemotherapeutic response by collagen gel droplet embedded culture drug sensitivity test in human breast cancers. AB - Collagen gel droplet embedded culture-drug sensitivity test (CD-DST) is the newly developed in vitro chemosensitivity test that has several advantages over the conventional ones. The aim of the present study is to examine the clinical usefulness of this test in the prediction of response to chemotherapy in breast cancer patients. Seventy patients with primary (n = 45) or locally recurrent (n = 25) breast cancers were recruited, and each patient underwent tumor biopsy before chemotherapy. The biopsy specimens were used for CD-DST and immunohistological examination of 6 biological markers (P-gp, erbB2, p53, BCL2, MIB1 and ER-alpha). As chemotherapy, cyclophosphamide 600 mg/m(2) plus epirubicin 60 mg/m(2) q3w (CE, n = 28) or docetaxel 60 mg/m(2) q3w (DOC, n = 42) was given. Interpretable results using the CD-DST assay were obtained from 84.3% (59/70) of tumor specimens studied. Of the 18 tumors diagnosed as CE sensitive by CD-DST, 15 (83.3%) exhibited a response to CE therapy and none of the 5 tumors diagnosed as CE resistant by CD-DST exhibited a response to CE therapy. Of the 14 tumors diagnosed as DOC sensitive by CD-DST, 13 (92.9%) exhibited a response to DOC therapy and only one of the 22 tumors diagnosed as DOC resistant by CD-DST exhibited a response to DOC therapy. P-gp expression was found to exhibit a significant (p < 0.05) association with the resistance to CE therapy but not to DOC therapy. Diagnostic accuracy (72.7%) achieved by P-gp was lower than that (87.0%) achieved by CD-DST in CE therapy. Expressions of other biological markers (erbB2, p53, BCL2, MIB1 and ER-alpha) were not significantly associated with response to CE or DOC therapy. These results demonstrate that CD-DST can predict the response to CE and DOC therapy with a high accuracy in breast cancer patients and seems to be superior to the conventional predictors. PMID- 11920600 TI - Efficient tumor targeting by single-domain antibody fragments of camels. AB - The variable domain of functional heavy chain antibodies (VHH) devoid of light chains, found in camels, constitute the smallest intact antigen-binding domain fragment. Two camel single-domain fragments, cAb-Lys2 and cAb-Lys3, recognizing an overlapping epitope of lysozyme with a dissociation constant of 2 nM and 65 nM, respectively, and a bivalent cAb-Lys3 were investigated for their ability to target transgenic tumors expressing lysozyme on their membrane. Biodistribution studies revealed that these non-immunogenic monomeric and bivalent camel single domain antigen binders specifically target lysozyme-expressing tumors and metastatic lesions. The excess of antibody is rapidly eliminated from the blood circulation and no cAb retention was observed in normal organs. The tumor to organ cAb-ratios at 2 and 8 hr were in the (2.1-10.8):1 and (6.2-23.7):1 range, respectively. The degree and specificity of tumor retention is independent of the affinity of the recombinant camel single-domain fragments for their antigen and from their univalent monomeric (15 kDa) or bivalent format (33 kDa). This study demonstrates the successful and specific in vivo targeting of tumors by camel single-domain fragments. It may open perspectives for their future use as tumor targeting vehicle, due to their small size, soluble behaviour and because they are non-immunogenic and interact with epitopes that are less antigenic for conventional antibodies. PMID- 11920602 TI - Interphase FISH assays for the detection of translocations with breakpoints in immunoglobulin light chain loci. AB - Many B-cell malignancies bear chromosomal translocations juxtaposing immunoglobulin (IG) genes with oncogenes, resulting in deregulated expression of the latter. Translocations affecting the IG heavy chain (IGH) locus in chromosomal region 14q32 are most prevalent. However, variant translocations involving the IG kappa (IGK) locus in 2p12 or the IG lambda (IGL) locus in 22q11 occur recurrently in B-cell neoplasias. No routine methods for the detection of all breakpoints involving IG light chain loci independently of the translocation partner have been described. For this reason, we have designed 2 novel interphase fluorescence in situ hybridization (FISH) assays using differentially labeled probes flanking the IGK and IGL locus, respectively. Based on extensive control studies, the diagnostic thresholds for the detection of breakpoints were set at 0.3% for IGK and 1.4% for IGL. Fifteen cases of B-cell malignancies with cytogenetically detectable chromosomal abnormalities in 2p11-14 were investigated with the FISH assay for IGK. Breakpoints affecting the IGK locus were detected in 7 cases including all 4 variant Burkitt's translocations t(2;8)(p12;q24) and a variant BCL2-associated translocation t(2;18)(p12;q21). Other translocation partners were chromosome bands 7q21 and 16q24. Ten cases with abnormalities in 22q11-12 were investigated with the FISH assay for IGL. Breakpoints in the IGL locus were diagnosed in 7 cases including both variant Burkitt's translocations t(8;22)(q24;q11) and a t(3;22)(q27;q11) involving the BCL6 locus. Other translocation partners were 2p13-14, 4q13 and 16p12. Our results show that these FISH assays provide flexible, simple and reliable tools in the diagnosis and characterization of genetic changes in B-cell malignancies. PMID- 11920601 TI - ZD1839 (IRESSA), an EGFR-selective tyrosine kinase inhibitor, enhances taxane activity in bcl-2 overexpressing, multidrug-resistant MCF-7 ADR human breast cancer cells. AB - Constitutive bcl-2 overexpression increases the tumorigenic and metastatic potential of doxorubicin-resistant, estrogen-independent, MCF-7 ADR human breast cancer cells. We evaluated the sensitivity to taxanes (paclitaxel, docetaxel and IDN 5109) of 2 bcl-2-overexpressing MCF-7 ADR clones and control neomycin transfected MCF-7 ADR neo cells. The 2 bcl-2-overexpressing MCF-7 ADR clones were relatively resistant to all 3 taxanes, whereas the MCF-7 ADR neo cells were relatively resistant to paclitaxel and docetaxel, but sensitive to IDN 5109. We found that both MCF-7 ADR neo and bcl-2-overexpressing MCF-7 ADR clones express high levels of the epidermal growth factor receptor (EGFR) and its ligand, transforming growth factor-alpha (TGF-alpha). Therefore, we tested the growth inhibitory effect of ZD1839 (Iressa, AstraZeneca, Macclesfield, UK), an orally active, selective EGFR tyrosine kinase inhibitor (EGFR-TKI) that is in clinical development. ZD1839 inhibited the growth in soft agar of all 3 clones in a dose dependent manner (IC(50) of approximately 0.1 microm). This effect was accompanied by a dose-dependent inhibition of EGFR tyrosine autophosphorylation and of the production of TGF-alpha, basic fibroblast growth factor (bFGF) and vascular endothelial growth factor (VEGF). To determine whether the blockade of EGFR signaling might affect the sensitivity of bcl-2-overexpressing MCF-7 ADR cells to taxanes, cells were treated with ZD1839 in combination with paclitaxel, docetaxel or IDN 5109, and dose-dependent cooperative growth inhibition as well as apoptosis potentiation were observed. Combined treatment with IDN 5109 and ZD1839 also resulted in a significant inhibition of bcl-2 expression in bcl-2 overexpressing MCF-7 ADR cells. These results demonstrate the ability of ZD1839 to overcome taxane resistance in a model of hormone-independent, multidrug resistant, human breast cancer. PMID- 11920603 TI - Cyclin D1 amplification in chromosomal band 11q13 is associated with overrepresentation of 3q21-q29 in head and neck carcinomas. AB - Eight cytogenetically characterized head and neck squamous cell carcinomas (HNSCCs) with CCND1 amplification in the form of a homogeneously staining region (hsr) in 11q13 were studied by COBRA FISH and FISH with specific probes to identify and characterize chromosomal segments added to the derivative chromosomes 11. In 4 of the tumors, it could be recognized that the material added was derived from the long arm of chromosome 3. The rearrangements were interpreted as der(11)hsr(11)(q13)t(3;11)(q21;q13) in 3 cases and as der(11)hsr(11)(q13)t(3;11)(q14;q13) in 1 case. In the other 4 cases, material from chromosomes 1, 16, or 19 was added to the derivative chromosomes 11. By further FISH analysis with 14 YAC clones spanning 3q13-q21 in the 4 tumors with der(11)hsr(11)t(3;11), it could be shown that they had different breakpoints at the molecular level, excluding the possibility that a particular gene was rearranged by the translocations. More surprisingly, gain of the 3q21-q29 segment was found in all 8 tumors with hsr in 11q13 and loss of 3p was seen in 7 of the tumors. These findings strongly indicate a synergistic effect of CCND1 amplification, loss of distal 11q, 3q gain and 3p deletion in HNSCC development and also suggests a mechanistic link between intrachromosomal amplification at 11q13 and recombination with distal 3q. PMID- 11920604 TI - Lifestyle factors and pancreatic cancer risk: a cohort study from the Swedish Twin Registry. PMID- 11920606 TI - Identification of cancer/testis genes by database mining and mRNA expression analysis. AB - Cancer/testis (CT) antigens are immunogenic proteins expressed predominantly in gametogenic tissue and cancer; they are considered promising target molecules for cancer vaccines. The identification of new CT genes is essential to the development of polyvalent cancer vaccines designed to overcome tumor heterogeneity and antigen loss. In the current study, a search for new CT genes was conducted by mining the Unigene database for gene clusters that contain expressed sequence tags derived solely from both normal testis and tumor-derived cDNA libraries. This search identified 1,325 different cancer/testis-associated Unigene clusters. The mRNA expression pattern of 73 cancer/testis-associated Unigene clusters was assessed by reverse transcriptase polymerase chain reaction. Three gene products, CT15/Hs.177959, CT16/Hs.245431 and CT17/Hs.178062, were detected only in testis and in tumor tissue. CT15 is equivalent to ADAM2/fertilin beta. CT16, an uncharacterized gene product, has homology (30-50%) to members of the GAGE gene family and is 89% identical to CT16.2/Hs.293317, indicating that CT16 and CT16.2 are members of a new GAGE gene family. The uncharacterized gene product, CT17, has homology (30%) to phospholipase A1. RT-PCR analysis showed that CT15 is expressed exclusively in renal cancer, whereas CT16 and CT17 are expressed in a range of human cancers. Real-time RT-PCR analysis of newly defined CT genes and the prototype CT antigens, MAGE-3 and NY-ESO-1, revealed low levels (less than 3% of the level detected in testis) of CT15, CT16 and NY-ESO-1 in a limited range of normal, non-gametogenic tissues. This study demonstrates the merits of database mining with respect to the identification of tissue-restricted gene products expressed in cancer. PMID- 11920607 TI - Effects of N-acetylcysteine in an esophageal carcinogenesis model in rats treated with diethylnitrosamine and diethyldithiocarbamate. AB - Due to the increasing role of esophageal tumors in human cancer pathology, there is need for animal models evaluating the mechanisms of esophageal carcinogenesis and investigating protective factors toward this disease. Several N-nitrosamines have been shown to induce esophageal tumors in rats. We designed a study in BD(6) rats treated with N-diethylnitrosamine (DEN) according to a simple protocol involving weekly i.p. injections of this carcinogen for 8 consecutive weeks. This treatment resulted in a high incidence and multiplicity of liver tumors and in occurrence of preneoplastic lesions and papillomas in the esophagus. Intraperitoneal injections of diethyldithiocarbamate (DEDTC), 4 hr after each DEN injection, i.e., during the period of DEN metabolization, improved survival of rats and did not affect the liver tumor yield but doubled the incidence of esophageal tumors and enhanced 4.9x their multiplicity. Moreover, 15% of rats developed esophageal squamocellular carcinomas. The oral administration of the thiol N-acetyl-L-cysteine (NAC), a precursor and analogue of reduced glutathione, to rats treated with the DEN/DEDTC combination did not change the liver tumor yield but attenuated esophageal carcinogenesis by producing a significant shift of preneoplastic lesions to milder forms as well as a significant decrease of tumor multiplicity. Therefore, the DEN/DEDTC protocol appears to provide an interesting 2-organ model of N-nitrosamine-induced carcinogenesis in rats, in which NAC is moderately effective as an inhibitor. The mechanisms underlying enhancement of DEN-induced esophageal carcinogenesis by DEDTC and the protective effects of NAC are discussed. PMID- 11920608 TI - Resistance of colon cancer cells to long-term 5-fluorouracil exposure is correlated to the relative level of Bcl-2 and Bcl-X(L) in addition to Bax and p53 status. AB - Defects in apoptosis have been implicated in chemoresistance of colon cancer cells. We report here the ability to resist to 5-fluorouracil-induced apoptosis of 8 colon cancer cell lines differing in p53 and bax status: p53(-/0)bax(+/+) for TC7, SW480, HT-29; p53(+/+)bax(-/-) for LS174T, LoVo; p53(+/+) bax(+/-) for HCT116; p53(+/+) or p53(+/0)bax(+/+) for LS513 or HCT-EB, respectively. To approximate to the in vivo therapy, the cell lines were exposed to a long-term treatment of 5-FU. The analysis of proteins implicated in the apoptotic pathway has shown that the independent analysis of p53 or bax status was not sufficient to predict the extent of drug-resistance of all cell lines. In p53(+/+) cell lines but not in p53(-/0) cell lines, a low level of the pro-apoptotic Bax protein was correlated with a greater resistance of cells to 5-FU. In addition, we found that high levels of anti-apoptotic Bcl-2 and Bcl-x(L) proteins combined with a low level of Bax were correlated to high 5-FU resistance of wild-type p53 cell lines. The same correlation was obtained for 2 out of 3 mutated p53 cell lines. In conclusion, the relative levels of Bcl-2, Bcl-x(L) and Bax may altogether contribute to determine the resistance of a majority of colon tumor cells to long-term 5-FU treatment, whatever their p53 status. PMID- 11920609 TI - Significance of the association between heparin-binding epidermal growth factor like growth factor and CD9 in human gastric cancer. AB - Heparin-binding epidermal growth factor-like growth factor (HB-EGF) is a member of the EGF family. Juxtacrine activity of proHB-EGF (the membrane-anchored form of HB-EGF) has been shown to be significantly potentiated when it is coexpressed with CD9 in vitro. The purpose of our study was to investigate the issue of whether proHB-EGF and CD9 are coexpressed in gastric cancer. HB-EGF gene expression and protein production in human gastric cancers was investigated, and EGF receptor and CD9 expressions were also evaluated. HB-EGF mRNA levels in gastric cancers were elevated, compared with normal gastric tissues, especially in the intestinal type. ProHB-EGF immunoreactivity was detected primarily in the cytoplasm and plasma membrane of gastric cancer cells. Of 66 patients, 40 (60.6%) exhibited proHB-EGF immunoreactivity and the level of its expression was significantly associated with tumor status (p < 0.01) and histological differentiation (p < 0.001). In addition, proHB-EGF mRNA was detected at high levels in the intestinal type by in situ hybridization. CD9 immunoreactivity was found to be preserved in 26 of 36 patients (72.2%) and CD9 protein expression was inversely associated with lymph node status (p < 0.05). A significant correlation between its expression and histological differentiation (p < 0.01) was found, and the association of CD9 with proHB-EGF was increased in the intestinal type, as evidenced by an immunoprecipitation method. These results indicate that the coexpression of proHB-EGF and CD9 may be involved in the tumorigenesis and/or proliferation of gastric cancers in a juxtacrine manner. PMID- 11920610 TI - Farnesyl thiosalicylic acid inhibits the growth of melanoma cells through a combination of cytostatic and pro-apoptotic effects. AB - Novel classes of drug that interfere with the signalling of the small G-protein Ras, the so-called Ras antagonists, are showing much promise as novel anti-cancer agents. In this study, we demonstrate that the novel Ras antagonist farnesylthiosalicylic acid (FTS) inhibits the growth of Colo 853 melanoma cells through a combination of cytostatic and pro-apoptotic effects. Furthermore, these phenomena are seen under conditions of cell attachment and in the presence of serum. Treatment of Colo 853 cells with FTS led to time-dependent inhibition of constitutive Akt, retinoblastoma protein (pRB) and ERK activity, with a concurrent loss of Akt expression. Inhibition of Akt and ERK activity induces apoptosis in other human cancer cell lines. Here it is demonstrated that inhibition of Akt, or ERK and Akt in combination, leads to cell cycle arrest but not apoptosis in melanoma cells. FTS treatment was also found to upregulate activity of the stress-activated p38 MAP kinase. Inhibition of p38 MAP kinase, using the selective inhibitor SB 203580, followed by FTS treatment, significantly increased the proportion of apoptotic cells after 72 hr, possibly suggesting a modulatory role for p38 MAP kinase in FTS-induced melanoma cell apoptosis. PMID- 11920611 TI - Proteome analysis of butyrate-treated human colon cancer cells (HT-29). AB - Butyrate, a 4-carbon fatty acid, has been shown to cause growth arrest and apoptosis of cancer cells in vitro and in vivo. The signaling pathways leading to changes in cell growth are unclear. We used a functional proteomics approach to delineate the pathways and mediators involved in butyrate action in HT-29 cells at 24 hr posttreatment. Using 2-dimensional gel electrophoresis, we showed that butyrate treatment resulted in alterations in the proteome of HT-29 cells. MALDI TOF mass spectrometry was used to identify butyrate-regulated spots. First, our results revealed that the expression of various components of the ubiquitin proteasome system was altered with butyrate treatment. This suggests that, in addition to the regulation of gene expression through the histone deacetylase pathway, proteolysis could be a means by which butyrate may regulate the expression of key proteins in the control of cell cycle, apoptosis and differentiation. Second, we found that both proapoptotic proteins (capase-4 and cathepsin D) and antiapoptotic proteins (hsp27, antioxidant protein-2 and pyruvate dehydrogenase E1) were simultaneously upregulated in butyrate-treated cells. Western blotting was carried out to confirm butyrate regulation of the spots. Both cathepsin D and hsp27 showed a time-dependent increase in expression with butyrate treatment in HT-29 cells. However, in HCT-116 cells, which were 5 fold more sensitive to butyrate-induced apoptosis, the upregulation of cathepsin D with time was not accompanied by a similar increase in hsp27 levels. Thus, the simultaneous upregulation of both proapoptotic and antiapoptotic proteins in HT 29 cells may account for their relative resistance to butyrate-induced apoptosis. PMID- 11920612 TI - Sex chromosome anomalies in pancreatic endocrine tumors. AB - We have investigated the status of sex chromosomes in 40 pancreatic endocrine tumors (PETs) using 2 complementary techniques: microsatellite and interphase FISH analysis. Twenty-five tumors were from female and 15 from male patients and included 31 nonfunctioning and 9 functioning PET (6 insulinomas, 2 glucagonomas and 1 VIPoma). Microsatellite and FISH analysis showed concordant results in all cases. PETs from females showed frequent loss of chromosome X (40%) whereas PETs from males showed relatively frequent loss of chromosome Y (36%) but never loss of the X chromosome. Statistical analysis showed significant association of sex chromosome loss with metastases (Spearman correlation test, r = 0.5, p < 0.001), local invasion (r = 0.33, p < 0.05) and high proliferation rate measured as Ki-67 index with a 5% cut-off (r = 0.42, p < 0.02). The analysis also showed that local invasion and metastases were highly correlated (r = 0.86). Multivariate survival analysis was therefore carried out including local invasion and loss of sex chromosomes. The presence of local invasion increased the risk of death almost 9 times whereas sex chromosome loss was an independent variable associated with a shorter survival period and an increased risk of death of approximately 4-fold. PMID- 11920613 TI - bc10: A novel human bladder cancer-associated protein with a conserved genomic structure downregulated in invasive cancer. AB - To identify novel genes that may be associated with the invasive phenotype of bladder cancer, we compared the mRNA expression profiles of fresh noninvasive (grade II, Ta) and invasive (grade III, T2-T4) human transitional cell carcinomas (TCCs) by mRNA differential display. Using this approach, we isolated a novel gene, designated bc10 (bladder cancer, Mr 10 kDa) that was exclusively expressed in the noninvasive lesions as judged by reverse transcriptase polymerase chain reaction analysis of a panel of 30 grade II, Ta and grade III, T2-T4 TCCs. The full-length bc10 cDNA contains a complete open reading frame (ORF) of 263 bp and encodes a protein composed of 87 amino acids that has no homology to any of the known protein families. Transient expression of bc10 cDNA in COS1 cells yielded a primary translation product with an apparent Mr of 9.8 kDa and pI of 6.7, in agreement with the theoretical calculated value. Comparison of mouse and human bc10 genomic loci revealed an intronless organization of the coding region in both species as well as a highly conserved structure having 91% and 100% identity at the DNA (coding region) and protein levels, respectively. Southern analysis did not reveal gross DNA rearrangements within the bc10 genomic locus in the invasive tumors, implying that the differential expression of the gene most likely reflects alterations in messenger expression (transcription and/or mRNA decay). The downregulation of this novel marker in invasive tumors suggests a putative role in bladder cancer progression. PMID- 11920614 TI - Expression profiling of MMA-1a and splice variant MMA-1b: new cancer/testis antigens identified in human melanoma. AB - Using high-density oligonucleotide array analysis, we have recently compared the gene expression profiles of 2 human melanoma cell lines with marked difference in metastatic behavior after subcutaneous inoculation into nude mice (de Wit et al., Melanoma Res, in press). We identified an expressed sequence tag (EST), which we called malignant melanoma-associated 1 (MMA-1a), showing evident differential expression between the 2 cell lines. The MMA-1a gene is localized on chromosome 21q22.2 and its mRNA exists of 4 exons. Homology search displayed a splice variant of MMA-1a that lacks exon 3 and that was called MMA-1b. Expression profiles of MMA-1a and MMA-1b are determined by reverse transciptase polymerase chain reaction (RT-PCR) analysis. Among 30 different normal tissue samples, expression of MMA-1a and MMA-1b was exclusively found in the testis after a first PCR of 30 cycles. Even more sensitive screening achieved by performing multiple semi-nested RT-PCR showed no or very low expression in the other normal tissues tested. During melanocytic tumor progression, MMA-1a and/or MMA-1b exhibited an emergence of expression in primary melanoma (20%) and melanoma metastasis samples (30%) after only 1 round of PCR. Expression of MMA-1a and/or MMA-1b was also identified in other tumor cell lines and fresh tumor samples of variable origin, e.g., lung, liver, bladder and soft tissues (sarcomas). We conclude that MMA-1a and MMA-1b are new members of the family of cancer/testis antigens. PMID- 11920615 TI - Immunotherapy with recombinant SFV-replicons expressing the P815A tumor antigen or IL-12 induces tumor regression. AB - Replicons based on alphaviruses are emerging as candidate vectors for vaccination and gene therapy purposes. We have reported previously that mice vaccinated with a recombinant Semliki Forest virus (a member of the alphavirus genus) carrying the gene encoding the P815A tumor antigen (rSFV/E-P1A) were protected against a lethal challenge with the P815 tumor. In this study we investigated the anti tumor therapeutic efficacy of rSFV/E-P1A or rSFV expressing the cytokine interleukin 12 (rSFV/IL12) on Day 5-established tumors. The results show that both antigen-specific and cytokine-mediated rSFV treatments exhibited a significant effect on P815 tumor growth, by delaying tumor progression and even inducing complete tumor regressions in several mice. The therapeutic potency of these vectors was dependent on the size of the treated tumor, as treatment of mice bearing larger tumors showed lower efficacy. In addition, rSFV treatment resulted in long-term immunity as observed by the lack of tumor recurrence in the majority of tumor-regressing mice after rechallenge with the tumor. Furthermore, the anti-tumor therapeutic effect was only achieved by local intratumoral injection of rSFV, as treatment by injection in the contralateral site resulted in tumor progression comparable to control-untreated mice. Accordingly, expression of a rSFV-RNA was localized to the tumor and draining lymph node. These results further demonstrate the potential of rSFV replicons as tumor therapeutic agents. PMID- 11920616 TI - Anti-viral cytotoxic T cells inhibit the growth of cancer cells with antibody targeted HLA class I/peptide complexes in SCID mice. AB - A number of experimental antibody mediated cancer therapies aim to redirect cytotoxic T cells (CTLs) of non-tumour specificity to cancer cells. It has been previously demonstrated that cancer cells targeted with recombinant HLA-class I/viral peptide complexes via antibody delivery systems can be killed by virus specific CTLs. This novel therapeutic system has been developed with a simple pre clinical model using the recombinant anti-CD20 B9E9 scFvSA fusion protein to target HLA-A2/peptide complexes to CD20 +ve Daudi lymphoma cells. In vitro data confirmed that, although binding of the B9E9 scFvSA fusion protein alone to Daudi cells had no effect on their growth, effective CTL mediated killing of Daudi cells could be achieved by targeting with B9E9 sfvScSA and recombinant HLA-A2/MI complexes at dilutions as low as 100 pg/ml. In contrast the free HLA-A2/MI complexes only significantly inhibited CTL activity at concentrations in excess of 100 ng/ml. The in vivo tumour protection assays in SCID mice demonstrated that only 1 of the 4 mice that received anti-HLA-A2/M1 CTLs and Daudi cells targeted with the B9E9 scFvSA fusion protein and HLA-A2/M1 complexes developed a tumour. In contrast in the control mice that received CTL and native Daudi cells all 4 developed tumours, as did all 4 that received targeted Daudi cells but no CTLs. Similar results were obtained in a parallel experiment using Daudi cells targeted with B9E9 scFvSA and HLA-A2/BMLF1 complexes and a CTL line to HLA-A2/BMLF1. The demonstration of in vivo activity for targeted HLA class I/peptide complexes combined with anti-viral T cells, supports the further clinical development of the system where it may be combined with autologous CTLs produced by vaccination or ex vivo expansion. PMID- 11920618 TI - Spontaneous hybridization of macrophages and Meth A sarcoma cells. AB - We present evidence of hybridization between Meth A sarcoma cells and syngeneic as well as semigeneic peritoneal macrophages. The resultant hybrids are characterized by morphology, membrane markers, ploidy, chromosomal content and functional features. Briefly, after a few days of coculture, cells appeared with morphology intermediate between the 2 original cell types. Typical macrophage surface molecules appeared in the hybrids. Meth A cells were labeled with red fluorescence and macrophages with green fluorescence. After 4 days in vitro, hybrids with yellow fluorescence appeared. Macrophages from BALB.K mice (H-2 K(k)) were cocultivated with Meth A cells from BALB/c mice (H-2 K(d)). The semigeneic hybrids displayed both specificities, as demonstrated by flow cytometry. The hybrids appeared moderately phagocytic, less so than the macrophages and markedly more so than the essentially nonphagocytic Meth A cells. The hybrids had a mean number of 76 chromosomes, as opposed to 53 in the Meth A cells and 40 in the macrophages. The macrophage DNA index was set at 1; Meth A cells were found to have an index of 1.6 in G1 phase, and the hybrids had a 2.6 index. The hybrids grew more slowly in vitro than Meth A cells, but grew faster in vivo. PMID- 11920617 TI - Infiltration of Th1 and Th2 lymphocytes around Hodgkin and Reed-Sternberg (H&RS) cells in Hodgkin disease: Relation with expression of CXC and CC chemokines on H&RS cells. AB - Hodgkin disease (HD) is characterized by the presence of Hodgkin and Reed Sternberg cells (H&RS) and a prominent lymphocytic infiltration. Various CXC and CC chemokines [e.g., interferon-gamma-inducible protein-10 (IP10), monokine induced by interferon-gamma (MIG) and TARC] are expressed on H&RS cells. Our study was designed to investigate the expression of MIG, IP10 and TARC on H&RS cells and their relations on lymphocyte infiltration. Immunohistochemical staining was performed using antibodies for CXCR3 (a specific receptor for IP10 and MIG), which is characteristic of Th1 helper phenotype and CCR3, CCR4 and ST2, which are features of Th2 cells. We studied 15 cases of HD [lymphocyte predominance (LP) type, 2; mixed cellularity (MC) type, 6; and nodular sclerosis (NS) type, 7]. All 6 MC cases contained TARC-, IP10- and MIG-expressing H&RS cells, however only 2 of 5 NS cases contained TARC-expressing H&RS cells, 3 of 7 NS contained MIG-expressing H&RS cells and only 1 of 7 NS contained IP10 expressing H&RS cells. Neither of the 2 LP cases contained HR&S cells that expressed these chemokines. The chemokines were more frequently expressed by MC than by NS and LP types. IP10-, MIG- and TARC-positive HD cases contained higher numbers of Th2 lymphocytes (ST2- CCR3- or CCR4-positive lymphocytes), compared to IP10-, MIG- and TARC-negative HD cases (p < 0.05 or <0.5). The number of CXCR3 (MIG and IP10 receptor)-positive lymphocytes (Th1 lymphocytes) was not different between MIG- and IP10-positive and -negative HD. Lymphocytes surrounding MIG- and IP10-positive H&RS cells were more frequently CXCR3-positive, however, compared to MIG- and IP10-negative cases. The CCR4 (TARC receptor)-positive lymphocytes, surrounding H&RS cells, were minority and the surrounding lymphocytes were not different between TARC-positive and negative cases. Our results indicate that MIG, IP10 and TARC chemokines influenced the response of Th2 cells in HD. It is possible, however, that IP10 and MIG may locally influence Th1 cells via cell migration. PMID- 11920619 TI - Extracellular matrix building marked by the N-terminal propeptide of procollagen type I reflect aggressiveness of recurrent breast cancer. AB - The purpose of our study was to examine the association between extracellular matrix homeostasis and aggressive breast cancer as reflected by the synthesis of type I collagen marked by circulating concentration of the aminoterminal propeptide of type I procollagen (PINP). Pre-therapeutic serum PINP concentrations were measured in 154 healthy women and 100 patients referred with their first metastatic manifestation of breast cancer and correlated to the metastatic pattern, response to therapy, time to progression and survival with a minimal follow-up of 5 years. Fifty-four percent of the patients had serum PINP concentrations greater than the 95th percentile of the healthy controls and 38% were high PINP level patients with values clearly outside normal range (>125 ng/ml). Patients with high PINP levels were more sick (p = 0.002), had a higher tumor burden (p = 0.013) and revealed a lower responsiveness to anthracycline based therapy (p = 0.0002) as well as an accelerated time to disease progression (p = 0.00001) and death (p = 0.0006). Median survival in the high serum PINP level group was less than half of that in the group with low PINP level (14.5 vs. 32 months). The lowest PINP levels were seen when the cancer was restricted to the lymph node and skin and increasing PINP levels were found if the cancer had spread to the lungs, the bones, the bone marrow and the liver. High PINP level at recurrence and lack of estrogen receptors (ER) independently reflected aggressive tumor behavior after recurrence with an equal great impact on time to progression and survival. Patients with a high PINP level and primarily ER-negative tumors survived a median of only 6 months with no one alive after 22 months. By contrast patients with a low PINP level and ER-positive tumors had a median survival of 37 months and 23% were still alive after 5 years. Aggressive breast cancer induces a strong fibroproliferative response with synthesis of type I collagen. Serum PINP levels may be a diagnostic and prognostic tool that indicate breast cancer activity, aggressiveness, expansion and metastasis and a predictor of outcome after anthracycline-based chemotherapy. PMID- 11920620 TI - Human papillomavirus 16 load in normal and abnormal cervical scrapes: an indicator of CIN II/III and viral clearance. AB - The relation between human papillomavirus type 16 (HPV 16) viral load in cervical scrapes and development of high-grade cervical intraepithelial neoplasia (CIN II or III) was studied in a nested case-control study of women with normal cytology (group A) and in a cohort of women with abnormal cytology (group B). HPV 16 DNA load was determined using a quantitative real-time PCR assay. In group A, case women (women with CIN II/III, n = 12) had a significantly higher viral load than control women (women with CIN < or = I, n = 47). This resulted in an increased relative risk of women with the 50% highest viral load for development of CIN II/III (OR 7.7; CI 1.6-33). In group B, women with CIN II/III (n = 38) had a significantly higher viral load than women with CIN < or = I (n = 25). Women with the 50% highest viral load had an increased relative risk of CIN II/III (OR 3.2; CI 1.1-9.3) and a decreased chance of both viral clearance and cytologic regression. Our data suggest that in women with normal cytology an increased HPV 16 load confers an increased risk of developing a CIN lesion. A sustained high viral load is subsequently informative for progression to a high-grade CIN lesion. PMID- 11920621 TI - BRCA1 and BRCA2 mutations among breast cancer patients from the Philippines. AB - Age-adjusted incidence rates of breast cancer vary more than 10-fold worldwide, with the highest rates reported in North America and Europe. The highest breast cancer incidence rates in Southeast Asia have been reported for the Manila Cancer Registry in the Philippines, with an age-standardized rate of 47.7 per 100,000 per year. The possible contribution of hereditary factors to these elevated rates has not been investigated. We conducted a case-control study of 294 unselected incident breast cancer cases and 346 female controls from Manila, Philippines. Cases and controls were selected from women below the age of 65 undergoing evaluation at the PGH in Manila because of a suspicious breast mass. Molecular analysis identified 12 BRCA2 mutations and 3 BRCA1 mutations. We estimate the prevalence of BRCA mutations among unselected breast cancer cases in the Philippines to be 5.1% (95% CI: 2.6-7.6%), with a prevalence of 4.1% (95% CI: 1.8 6.4%) for BRCA2 mutations alone. The BRCA2 4265delCT and 4859delA mutations were found in 2 and 4 unrelated cases, respectively; haplotype analysis confirmed that these, and the BRCA1 5454delC mutation, are founder mutations. BRCA2 mutations were also found in 2 of 346 controls (0.6%; 95% CI: 0.2-1.4%). Compared with non carrier cases, the cumulative risk of breast cancer for first-degree relatives of mutation carriers was 24.3% to age 50, compared with <4% for first-degree relatives of non-carrier cases (RR = 6.6; 95% CI: 2.6-17.2; p= 7.5 x 10(-6)). Our data suggest that penetrance of BRCA mutations is not reduced in the Philippines. Germline mutations in the BRCA2 gene contribute more than mutations BRCA1 to breast cancer in the Philippines, due in large part to the presence of 2 common founder mutations. PMID- 11920622 TI - Breast cancer risk factors and age at diagnosis: an Icelandic cohort study. AB - An increasing number of studies indicates that the strength and even direction of association between breast cancer and established risk factors differ according to the woman's age when she develops the disease. This was studied in the setting of a population based cancer registry using a databank with information on age at menarche, parity, age at first birth, oral contraceptive (OC) use, lactation, height and weight. From a cohort of 80.219 women attending population-based cervical and breast cancer screening in Iceland, 1120 cases were identified, aged 26-90 years at diagnosis and 10,537 controls, individually matched to the cases on birth year and age when attending. Information given at last visit before diagnosis was used in the analysis, applying conditional logistic regression. Odds ratios and statistical strength of relationships varied according to age at diagnosis for age at first birth, number of births, duration of lactation, height and weight. The decreased risk associated with young age at first birth and increasing duration of breast feeding became less pronounced with advancing age at diagnosis. A reduced risk associated with an increasing number of births was not detected in women diagnosed under the age of 40. An increased risk associated with giving first birth after 30 years of age was mainly detected in women who had only given 1 birth and were diagnosed under the age of 40 (OR = 7.06 95% CI = 2.16-23.01). A positive association with height and especially with weight was confined to women diagnosed after the age of 55. The results confirm that age at diagnosis should be taken into account when studying the effects of breast cancer risk factors. PMID- 11920623 TI - History of allergies among adults with glioma and controls. AB - The causes of most adult gliomas are essentially unknown. Previous studies have indicated associations between immune system factors and the incidence of adult glioma, specifically that those individuals with certain allergic conditions may have decreased risk of glioma. We obtained detailed allergy histories for 405 adults newly diagnosed with glioma in the San Francisco Bay Area from 1997-1999 and 402 age-gender-ethnicity frequency-matched population-based controls. Seventy nine percent of eligible cases or their proxies and 74% of eligible controls completed in-person interviews about allergies, age at onset, frequency, duration and severity. Overall, cases were less likely than controls to report any allergy (72% vs. 85%; odds ratio [OR] = 0.5 [0.3-0.7]); for self-reported cases (n = 269), OR = 0.7 (0.4-0.97) and for proxy-reported cases, OR = 0.3 (0.2-0.5). Pollen, dairy and nut allergies were significantly less common in cases than controls and most other allergens had odds ratios of less than one. There were no apparent trends with numbers of symptoms, route of exposure of allergen or reported severity of allergy, but there was a significant dose-response with increasing numbers of allergens (p < 0.0001 for linear trend among all cases vs. controls and p = 0.02 among self-reported cases only vs. controls). Although our work displays strong and consistent associations, future efforts must attempt to establish whether an immune system typified by proclivity to allergies, or an immunologic consequence of the allergies themselves, might be capable of preventing nascent brain tumors. The dominance of humoral immunity in the central nervous system is consistent with either of these models. Alternatively, common genetic or environmental causes for allergies and gliomagenesis may mediate or confound these observed inverse risks for allergies and gliomas, or other explanations may exist. Future work might reveal an important role for immunologic factors in gliomagenesis and potential preventative and/or therapeutic modalities. PMID- 11920624 TI - Cyclopentenyl cytosine increases the phosphorylation and incorporation into DNA of 1-beta-D-arabinofuranosyl cytosine in a human T-lymphoblastic cell line. AB - The cytotoxic effect of 1-beta-D-arabinofuranosyl cytosine (araC) depends on the intracellular phosphorylation into its active compound araCTP, on the degree of degradation of araCTP and on the incorporation of araCTP into DNA. Deoxycytidine triphosphate (dCTP) inhibits the phosphorylation of araC (by feedback inhibition of the enzyme deoxycytidine kinase) and the incorporation of araCTP into DNA (by competition for DNA polymerase). In a T-lymphoblastic cell line, we studied whether the cytotoxicity of araC (2 nM-50 microM) could be enhanced by decreasing the concentration of dCTP, using the nucleoside-analogue cyclopentenyl cytosine (CPEC), an inhibitor of the enzyme CTP synthetase. Preincubation of the cells with CPEC (100-1,600 nM) for 2 hr increased the concentrations of araCMP 1.6-9.5 fold, which was significant for each concentration of CPEC used. The concentration of araCDP remained low, whereas the concentration of araCTP changed depending on the concentration of araC used. With 2-15 microm of araC and a preincubation with 400 nM of CPEC, the araCTP concentration increased by 4-15% (not significant), and the total amount of araC nucleotides increased significantly by 21-45%. When using a concentration of araC of 2 nM after a preincubation with CPEC of 100 nM, the concentration of araCMP increased by 60% (p = 0.015), whereas that of araCTP decreased by 10% (p = 0.008). This was compensated by an increase of 41% (p = 0.005) of araCTP incorporation into DNA, which represented 43% of all araC metabolites. Moreover, by performing pulse/chase experiments with 400 nM of CPEC and 2 microM of araC, the retention of cytosolic araCTP and the incorporated amount of araCTP into DNA were increased by CPEC. The modulation by CPEC of araC metabolism was accompanied by a synergistic increase of araC-induced apoptosis and by an additive effect on the araC-induced growth inhibition. PMID- 11920625 TI - Inhibition of PC-3 human prostate cancers by analogs of growth hormone-releasing hormone (GH-RH) endowed with vasoactive intestinal peptide (VIP) antagonistic activity. AB - Vasoactive intestinal peptide (VIP) stimulates the proliferation and invasiveness of malignant prostatic cells. Receptors for VIP and the closely related growth hormone-releasing hormone (GH-RH) show considerable homology and are found in prostatic and other carcinomas. Among various analogs of GH-RH synthesized, JV-1 52 is a non-selective VIP/GH-RH antagonist, whereas JV-1-53 is a VIP antagonist devoid of GH-RH antagonistic effect. In our study, nude mice bearing PC-3 human androgen-independent prostate carcinomas were treated with JV-1-52 or JV-1-53 (20 microg/day, s.c.) for 28 days. Both antagonists produced a similar reduction in tumor volume (62-67%, p < 0.01) and tumor weight (59-62%; p < 0.05) vs. controls and extended tumor doubling-time from 9.1 to about 16 days (p < 0.05). To investigate the mechanisms involved, in another study we compared the effects of JV-1-53 with those of somatostatin analog RC-160. VIP antagonist JV-1-53 reduced tumor weight by 67% (p < 0.01) and suppressed the expression of mRNA for c-fos and c-jun oncogenes by about 34% (p < 0.05), without affecting serum levels of insulin-like growth factor-I (IGF-I). In contrast, RC-160 (50 microg/day) reduced serum IGF-I by 19% (p < 0.05), but did not significantly decrease tumor weight. mRNA for VIP and high affinity receptors for VIP were detected on PC-3 tumors. Our results suggest that VIP/GH-RH antagonists can inhibit the growth of androgen independent prostate cancer by abrogating the autocrine/paracrine mitogenic stimuli of VIP. The ability of GH-RH antagonists to block tumoral VIP receptors, in addition to GH-RH receptors, could be potentially beneficial for prostate cancer therapy. PMID- 11920626 TI - Increased expression of multidrug resistance-associated proteins in bladder cancer during clinical course and drug resistance to doxorubicin. AB - Overexpression of the P-glycoprotein/multidrug resistance 1 (MDR1) and multidrug resistance protein 1 (MRP1) gene is closely associated with the clinical outcome of various malignancies, and it is involved in responses to some anticancer chemotherapeutic agents including doxorubicin. Six human MRP subfamily members (MRP2-7) with structural similarities to MRP1 have been identified. Recently, the relationships between MRP2 and MRP3 expression levels of some cancer cells and drug sensitivity to doxorubicin have been reported, but the relationship between the clinical samples and drug sensitivity remains unclear. We determined the expressions of the MDR1, MRP1, MRP2 and MRP3 gene in bladder cancer during the clinical course and sought to learn whether the expression was correlated with drug responses to doxorubicin. Doxorubicin, used in chemotherapeutic treatment including intravesical and systemic chemotherapy, is an important anticancer agent for the treatment of bladder cancer. We used quantitative reverse transcriptase-polymerase chain reaction (RT-PCR) analysis for our study, and the sensitivity to doxorubicin in bladder cancer was determined using the in vitro succinate dehydrogenase inhibition test. Using 47 clinical samples of bladder cancer, we confirmed the significant correlation of MDR1, MRP1 and MRP3 mRNA levels with resistance to doxorubicin. We showed that the expression of MDR1, MRP1, MRP2 and MRP3 in recurrent tumors and residual tumors after chemotherapeutic treatment was higher than that in untreated primary tumors. In particular, the MDR1 expression in residual tumors was 5.7-fold higher than that in untreated primary tumors. PMID- 11920627 TI - Cancer mortality in the European Union, 1988-1997: the fall may approach 80,000 deaths a year. AB - After the peak rate of cancer mortality reached in 1988 in the European Union, steady declines were observed: 9.1% for both sexes combined over the period 1988 1997 (from 147.0 to 133.6/100,000, world standard), corresponding to the avoidance of about 80,000 deaths in 1997 (approximately 39,000 below age 65 and 41,000 above). In 1997, the total number of cancer deaths also declined, for the first time. The major determining cancers for these favorable trends were stomach (-30%), lung (-10%), intestines (-15%), breast (-10%), uterus (mainly cervix; 22%), leukemias (-10%) and, after 1995, prostate (-3%). PMID- 11920628 TI - No mutations in the BACH1 gene in BRCA1 and BRCA2 negative breast-cancer families linked to 17q22. PMID- 11920629 TI - Expression of SSX genes in human osteosarcomas. PMID- 11920631 TI - Use of H19 regulatory sequences for targeted gene therapy in cancer. AB - We present a tumor gene therapy approach based on the use of regulatory sequences of the H19 gene that are differentially expressed between normal and cancer cells. We constructed expression vectors carrying the gene for the A fragment of diphtheria toxin (DT-A) or herpes simplex virus thymidine kinase (HSV-tk), under the control of a 814 bp 5'-flanking region of the H19 gene. The cell killing activity of these constructs was in accordance with the relative activity of the H19 regulatory sequences in the transfected cells. We evaluated the therapeutic potential of the gene expression constructs driven by H19 regulatory sequences in an animal model of bladder cancer induced by subcutaneous injection of syngeneic bladder tumor cell lines. Intratumoral injection of these constructs caused a significant suppression of subcutaneous tumor growth, with no obvious toxicity toward the host. PMID- 11920632 TI - Promoter hypermethylation of the EDNRB gene in nasopharyngeal carcinoma. AB - To identify the epigenetic changes in nasopharyngeal carcinoma (NPC), we performed methylation-sensitive restriction fingerprinting (MSRF) analysis on NPC cell lines and xenografts. A 190 bp sequence methylated in NPC tumors was isolated and showed high homology to the 5' CpG island of the endothelin receptor B (EDNRB) gene. Since the EDNRB gene is commonly inactivated in prostate and bladder cancers, it may be a candidate target gene involved in NPC tumorigenesis. By bisulfite sequencing, we have confirmed that hypermethylation of the 5' CpG island of EDNRB occurred in both xenografts and all 4 cell lines but not in 2 normal nasopharyngeal outgrowths. RT-PCR demonstrated that only original EDNRB transcripts, but not the splicing transcripts, were expressed in normal nasopharyngeal epithelial cells. Loss of the original EDNRB expression was consistently found in 2 xenografts and 3 cell lines with dense methylation patterns. Treatment of these 3 cell lines with 5'-aza-2'-deoxycytidine led to re expression of the EDNRB transcript and demethylation of its promoter regions. Our results demonstrate that silencing of EDNRB gene expression in NPC is associated with promoter hypermethylation. Using methylation-specific PCR, we also detected methylation of the 5' CpG island of EDNRB in 19/21 (90.5%) primary tumors, while no methylation was found in all 6 normal nasopharyngeal epithelia. The high frequencies of promoter hypermethylation suggest that repression of the EDNRB gene may play a role in the development of NPC. PMID- 11920633 TI - Urokinase receptor-associated protein (uPARAP) is expressed in connection with malignant as well as benign lesions of the human breast and occurs in specific populations of stromal cells. AB - The urokinase-type plasminogen activator (uPA) and the uPA receptor (uPAR) are key components in the plasminogen activation system, serving to promote specific events of extracellular matrix degradation in connection with tissue remodeling and cancer invasion. We recently described a new uPAR-associated protein (uPARAP), an internalization receptor that interacts with the pro-uPA:uPAR complex. In our study, we generated a specific polyclonal peptide antibody against human uPARAP and used it for the localization of uPARAP in different breast lesions. The affinity-purified antibodies specifically recognized uPARAP in Western blotting and gave a strong signal in immunohistochemistry. The immunohistochemic localization pattern was found to be identical to that of uPARAP mRNA as determined in parallel by in situ hybridization. uPARAP expression was then studied in both benign and malignant breast lesions. Whereas the normal breast tissue was uPARAP-negative, all benign lesions and ductal carcinoma in situ lesions showed immunoreactivity in fibroblast-like cells and myoepithelial cells associated with the lesion. In invasive carcinoma, uPARAP immunoreactivity was limited to tumor-associated mesenchymal cells. Double immunofluorescence analysis of invasive ductal carcinoma using antibodies against specific cell markers showed that uPARAP was localized in myofibroblasts and macrophages. No malignant cells, no endothelial cells and no vascular smooth muscle cells showed uPARAP immunoreactivity. We conclude that expression of uPARAP is associated with the abnormal breast and that expression appears in myofibroblasts, macrophages and myoepithelium. We suggest that uPARAP is involved in the clearance of the uPA:uPAR complex as well as other possible ligands during benign and malignant tissue remodeling. PMID- 11920634 TI - Insulin-like growth factor-I has a biphasic effect on colon carcinoma cells through transient inactivation of forkhead1, initially mitogenic, then mediating growth arrest and differentiation. AB - IGF-I stimulates intestinal cell differentiation after initiating a short proliferative burst, similar to its effect on muscle cell differentiation. Levels of IGF-I attainable in serum (10-20 ng/ml) induced transient growth stimulation of colon carcinoma cells, then growth arrest. When IGF-I functioned as a mitogen, it blocked differentiation. Intestinal cell differentiation occurred once cells had undergone the IGF-I-initiated growth arrest and IGF-I and butyrate acted synergistically to induce maturation markers. IGF-I induces NIH-3T3 cell proliferation and survival by activating the kinase akt, which in turn inhibits various apoptotic mediators and the forkhead family of transcription factors, which mediate expression of p27(kip1). Promoter reporter assays demonstrated that forkhead1 mediates transcription of p27(kip1) in colon carcinoma cells. The mitogenic effects of IGF-I on 4 colon carcinoma cell lines were transient because the inactivating phosphorylation of forkhead1 by akt was short-lived. This allowed transcriptional upregulation of the cdk inhibitor p27(kip1), with a resulting growth arrest. In contrast, in NIH-3T3 cells treated in parallel with identical IGF-I levels, forkhead phosphorylation levels were sustained; thus, no increase in p27(kip1) levels was seen and cells continued to proliferate. Intestinal epithelial cells in vivo undergo a limited number of divisions, then growth arrest and completion of their maturation. IGFs found in intestinal tissue may control the timing of this process. In addition, colon cancers may have developed strategies to overcome IGF-I-mediated growth arrest. Earlier (Kansra et al., Int J Cancer 2000;87:373-8), we found that levels of IGFBP-3 were elevated at least 2-fold in 70% of resected colon cancers compared with adjacent normal tissue. In the current study, growth inhibition by IGF-I and IGF-II was blocked by concurrent addition of IGFBP-3, implying that colon cancers with elevated IGFBP-3 levels would be selected for in vivo because they could bind and inactivate high serum IGF-I levels and continue to proliferate. PMID- 11920635 TI - Role of low nuclear grading of renal carcinoma cells in the functional profile of tumor-infiltrating T cells. AB - Tumor-infiltrating lymphocytes (TILs) from biopsies of 9 selected patients with pT1pN0M0 renal cell carcinoma (RCC) were analyzed at the clonal level for phenotypic distribution, cytokine secretion profile and antitumor cell proliferation and cytotoxicity. T-cell clones generated from RCCs were able to produce higher amounts of interleukin-4 (IL-4) and interferon-gamma (IFN-gamma) than the corresponding clones derived from peripheral blood mononuclear cells, thus suggesting a recruitment into tumors of T cells with peculiar functions. Moreover, CD4+ T-cell clones generated from TILs of nuclear grading 2 (G2)-type RCC patients produced significantly higher amounts of IL-4 and IL-10 and lower amounts of IFN-gamma than the corresponding clones generated from G1-type RCC and 2 renal angiomyolipoma (AML) patients. In addition, T-cell clones generated from lymphocytes infiltrating the peritumoral areas of G2-type, but not those from G1 type, RCC patients produced higher and lower amounts of IL-4 and IFN-gamma, respectively, than the corresponding clones derived from intratumoral T cells of the same patients. The proportion of T-cell clones derived from G2-type tumors and proliferating to autologous tumor cells (ATCs) was significantly lower than that of clones generated from G1-type RCC or AML patients. However, irrespective of their source, they exhibited similar cytokine profiles and produced comparable amounts of IL-4, IL-10 and IFN-gamma. Furthermore, the proportion and the production of both IL-4 and IFN-gamma of G2-type RCC-derived T-cell clones with cytotoxic activity against ATC were significantly lower than those of cytolytic clones generated from AML and G1-type RCCs. The concentrations of IL-4, IL-10 and IFN-gamma produced by the cytolytic clones from G2-type RCC were also lower than those produced by their noncytolytic counterparts obtained from the same patients. These data address the association of the nuclear grading of neoplastic cells with different local tumor-specific T-cell responses in RCC. PMID- 11920636 TI - Inhibition of tumor growth by systemic treatment with thrombospondin-1 peptide mimetics. AB - Many normal human cells produce thrombospondin-1 (TSP-1), a potent antiangiogenic protein that promotes vascular quiescence. In various organ systems, including the brain, breast and bladder and in fibroblasts, TSP-1 secretion is reduced during tumorigenesis, thereby allowing induction of the vigorous neovascularization required for tumor growth and metastasis. Full-length and short TSP-1-derived peptides inhibit angiogenesis by inducing endothelial cell apoptosis and thus disrupting the vasculature of the growing tumor. CD36 expressed on the surface of endothelial cells functions as the primary antiangiogenic receptor for TSP-1. A D-isoleucyl enantiomer of a TSP-1 heptapeptide specifically inhibits the proliferation and migration of capillary endothelial cells. DI-TSP, an approximately 1 kDa capped version of this peptide, is also antiangiogenic in vitro, with a specific activity approaching that of the 450 kDa parental molecule. Here, we show that DI-TSP delivered systemically dose dependently inhibits the growth of murine melanoma metastases in syngeneic animals and that its more soluble isomer, DI-TSPa, similarly blocks the progression of primary human bladder tumors in an orthotopic model in immune deficient mice. Like intact TSP-1, these peptide mimetics had no effect on cancer cells growing in vitro but markedly suppressed the growth of endothelial cells by inducing receptor-dependent apoptosis. Antibodies raised against CD36 blocked the ability of peptides to induce apoptosis in endothelial cells but had no effect on tumor necrosis factor-alpha-induced apoptosis. In vivo, the peptide mimetics were associated with a significantly reduced microvessel density and increased apoptotic indices in both the endothelial and tumor cell compartments. Such short peptides targeted to a specific antiangiogenic receptor, potent and easy to synthesize, show great promise as lead compounds in clinical antiangiogenic strategies. PMID- 11920637 TI - alpha v-Integrin antagonist EMD 121974 induces apoptosis in brain tumor cells growing on vitronectin and tenascin. AB - Orthotopic brain tumor growth is inhibited in athymic mice by the daily systemic administration of the alpha v-integrin antagonist EMD 121974. This compound, a cyclic RGD-penta-peptide, is a potent inhibitor of angiogenesis, which induces apoptosis of growing endothelial cells through inhibition of their alpha v integrin interaction with the matrix proteins vitronectin and tenascin. Here we show that EMD 121974 also induces apoptosis in the alpha v-integrin-expressing tumor cell lines U87 MG and DAOY by detaching them from vitronectin and tenascin, matrix proteins known to be essential for brain tumor growth and invasion. These matrix proteins are shown to be produced by the brain tumor cells in vitro and in vivo. Furthermore, only tumor cells expressing alpha v-integrins responded to the treatment with EMD 121974, after xenotransplantation into the forebrain of nude mice, supporting the importance of tumor cell-matrix interactions in tumor cell survival in the brain. Thus, the alpha v-antagonist EMD 121974 suppresses brain tumor growth through induction of apoptosis in both brain capillary and brain tumor cells by preventing their interaction with the matrix proteins vitronectin and tenascin. The dual action of this peptide explains its potent growth suppression of orthotopically transplanted brain tumors. PMID- 11920638 TI - Effect of prolonged hydroxytamoxifen treatment of MCF-7 cells on mitogen activated kinase cascade. AB - Resistance to the antiestrogen tamoxifen is the main stumbling block for the success of breast cancer therapy. We focused our study on cellular alterations induced by a prolonged treatment with the active tamoxifen metabolite hydroxytamoxifen (OHT). We show that a prolonged OHT treatment (for up to 7 days) led to a progressive increase in the level of phosphorylated p44/42 mitogen activated kinase (MAP kinase) induced by 10(-7) M TPA stimulation, without any significant change in the protein level. This effect was also observed in MCF-7 cells grown first in medium containing dextran-coated charcoal-treated FCS (DCC medium) for 20 days prior to OHT treatment, indicating a specific effect of the antiestrogen and not an effect of estrogen deprivation. It was prevented by cotreatment with estradiol and not observed in the estrogen receptor negative HeLa cell line, suggesting that it was mediated by the estrogen receptor. TPA induced phosphorylation of MEK1/2 was also raised by OHT treatment, without any change in their protein level or Raf-1 and H-Ras levels. When the MCF-7R OHT resistant cell line was grown in antiestrogen containing medium, the level of phosphorylated p44/42 MAP kinase was also high but reversed when the antiestrogen was removed. The 2 other MAP kinase, JNK and P38 pathways were not affected in the same way by OHT treatment. In conclusion, our data reveal that a prolonged OHT treatment, by increasing p44/42 MAPK activity, affects a key step in the growth control of MCF-7 cells, although not sufficiently to overcome the growth inhibitory effect of the drug. PMID- 11920639 TI - Osteopontin induces angiogenesis of murine neuroblastoma cells in mice. AB - Angiogenesis is an essential process for tumor growth and is regulated by tumor derived angiogenic cytokines. Osteopontin (OPN) is one of the cytokines produced by various tumor cells and is suggested to be involved in angiogenesis by upregulating endothelial cell migration in cooperation with vascular endothelial cell growth factor (VEGF). To provide evidence of OPN involvement in a causal role in tumor angiogenesis, we generated a stable transfectant from murine neuroblastoma C1300 cells to constitutively secrete high levels of murine OPN. The OPN mRNA expression and protein secretion were confirmed by RT-PCR and ELISA, respectively. The biological activity of secreted OPN was determined with migration assay by using human umbilical vein endothelial cells (HUVEC). Transfection with OPN gene did not increase VEGF production and did not affect gene expression of other angiogenic factors confirmed by complementary DNA macroarray system. To demonstrate the effect of OPN on tumor-induced angiogenesis in vivo, millipore chambers containing OPN-transfected or control cells were implanted to the dorsal air sac of mice. The OPN-transfected cells significantly induced neovascularization in comparison to the control cells in mice. Conclusively, these results provide direct evidence of OPN involvement in the role of tumor angiogenesis. PMID- 11920640 TI - Different pattern of loss of heterozygosity among endocervical-type adenocarcinoma, endometrioid-type adenocarcinoma and adenoma malignum of the uterine cervix. AB - It is unclear which chromosome arms frequently show loss of heterozygosity (LOH) in adenocarcinoma of the uterine cervix. To identify such chromosomal arms, LOH on 52 different chromosome loci was examined using laser capture microdissection and PCR-LOH analysis in 25 common-type adenocarcinomas, comprising 13 cases of endocervical type, 12 cases of endometrioid type and 7 cases of adenoma malignum without the component of conventional endocervical-type adenocarcinoma (designated as "pure" form). In adenocarcinomas of endocervical type and endometrioid type, LOH was commonly detected on chromosome arms 17p (62% and 50%, respectively), 1p (33% and 67%) and 22q (40% and 33%). In addition, endocervical type adenocarcinoma frequently (> or = 30%) showed LOH on 18p (71%), 19q (50%), 19p (38%) and 16q (38%), whereas endometrioid-type adenocarcinoma frequently showed LOH on 10q (43%) and 5q (40%). LOH was only sporadically detected on 9q, 18q, or 21q in 3 of 7 cases of "pure" adenoma malignum. In a case of coexistence of "pure" adenoma malignum and adenocarcinoma in situ (AIS), LOH on 1q, 5p, 11p, 17p, 18p and 18q was detected only in AIS. LOH was accumulated on a number of chromosome arms in the adenocarcinoma at the early developmental stage before stromal invasion. Chromosomal arms that are prone to show LOH appeared to differ between the 2 types of cervical adenocarcinoma. We could suggest that "pure" adenoma malignum is of clonal and neoplastic nature in view of the detection of LOH. PMID- 11920641 TI - Expression and mutation analyses of P53R2, a newly identified p53 target for DNA repair in human gastric carcinoma. AB - p53R2, a recently identified putative tumor suppressor located at 8q23.1, encodes a protein with striking similarity to a small subunit of ribonucleotide reductase. p53R2 is directly induced by wild-type p53 and involved in the p53 checkpoint for repair of damaged DNA, raising the possibility that mutational inactivation of p53R2 may contribute to the development and progression of human malignancies. To explore the p53R2's candidacy for a suppressor in gastric tumorigenesis, we examined the expression and mutation status of p53R2 in 166 gastric specimens including 90 primary adenocarcinomas and 15 cell lines. In response to genotoxic damages, p53R2 transcription was clearly activated in wild type but not mutant-type p53-carrying cells while basal expression of p53R2 in undamaged cells showed no association with the mutational status of p53. Host cell reactivation assay revealed that p53R2 enhances DNA repair efficiency and plays a role in the p53-mediated repair of damaged DNA, whereas no significant effect of p53R2 on cell growth and apoptosis was detected in flow cytometry and [(3)H]thymidine incorporation assays. p53R2 transcript was expressed in all normal and tumor tissues and its expression levels were not significantly different between normal and malignant carcinoma tissues. p53R2 expression showed no correlation with stage, grade and histological types of tumors. Moreover, no tumor-specific reduction of p53R2 was detected in 30 matched sets. Mutational analysis of p53R2 in 105 carcinomas including 15 cell lines also failed to detect any evidences for genomic deletion or somatic mutations leading to amino acid substitutions or frameshift whereas 31% (28 of 90) of the same primary tumors showed p53 alterations. Whereas 82% (23 of 28) of the mutant p53-carrying primary tumors expressed abnormally low p21(Waf1), no association of p53R2 expression with the p53 status was recognized, suggesting that basal transcription of p53R2 is regulated through the p53-independent mechanism. Collectively, our study indicates that although p53R2 is induced in a p53-dependent manner and involved the p53-mediated DNA repair in gastric epithelial cells, it is not a critical target of genetic inactivation in gastric tumorigenesis. PMID- 11920642 TI - Alterations of the p16(ink4a) gene in resected nonsmall cell lung tumors and exfoliated cells within sputum. AB - Recently, we reported that p16 protein expression was nondetectable in 49.5% of 107 resected nonsmall cell lung cancers (NSCLCs), suggesting that the p16(INK4a) gene is frequently inactivated in primary NSCLC. To identify the molecular basis for this p16 immunohistochemical negativity further, we performed a genetic and epigenetic study of p16(INK4a) status in a series of 115 NSCLC samples parallel to the clinicopathologic and prognostic analyses. Microdissected tumor DNA samples were screened for homozygous deletion using comparative multiplex polymerase chain reaction (PCR), for intragenic mutation using direct sequencing and for loss of heterozygosity (LOH) using an intragenic microsatellite marker, D9S942. Of these samples, 67 were further analyzed by SmaI-based PCR methylation assay to evaluate aberrant methylation at the gene. To examine the correlation of aberrant methylation in tumor and sputum samples, sputum samples from 12 matched patients were assessed for this change. We found that methylation of the p16(INK4a) gene was present in 38 of the 67 (56.7%) tumors and was significantly associated with negative p16 protein expression (p = 0.029). A 92% (11/12) concordance of sputum samples with matched resected tumors was found. The survival rates among adenocarcinoma patients with p16(INK4a) methylation were lower, but at a level of borderline significance compared with those patients without methylation (p = 0.071). In addition, 29.4% of the informative cases were found to harbor LOH at D9S942. None of the 115 microdissected tumors exhibited homozygous deletion in the p16(INK4a) gene. Only 1 patient exhibited a complex mutation at the fourth ankyrin repeat consensus sequence and concordantly demonstrated p16 immunohistochemical negativity. Overall, 69% (79/115) of NSCLC tumors had at least 1 type of p16(INK4a) alteration. Our data provide compelling evidence that p16(INK4a) alterations are involved in NSCLC tumorigenesis and that promoter methylation is the predominant mechanism in p16(INK4a) deregulation. PMID- 11920643 TI - Identification of BRCA1 and BRCA2 carriers by allele-specific gene expression (AGE) analysis. AB - Mutations in BRCA1 and BRCA2 genes confer a high risk of breast and ovarian cancer. As such, their identification is essential to reduce the risk of disease in healthy carriers, as well as in carriers who have already developed the disease because they are at increased risk for a second malignancy; moreover, noncarriers of BRCA1 and BRCA2 mutated families can be spared anxiety and unnecessary medical interventions. A number of problems, including large gene size, complex mutational spectra and genetic heterogeneity of the disease, however, make genetic testing labor intensive and often inconclusive. We devised a new mutation detection strategy called AGE (allele-specific gene expression) analysis that relies on the detection of a "functional effect" of the mutation at the RNA level known as "nonsense-mediated RNA decay," thus avoiding several of the problems of BRCA1 and BRCA2 genetic testing. In particular, (i) AGE analysis discriminates among the predisposing genes and identifies mutation carriers with a single RT-PCR reaction; (ii) it relies on the effect of truncating mutations, which represent the large majority of cases and thus identifies mutation carriers regardless of the specific genomic alteration; and (iii) it is specific for cis regulatory mutations that are missed at present by most of the methods. As AGE analysis has the potential to identify most of the BRCA1 and BRCA2 mutation carriers, it can be used as a preliminary screening method, thereby accelerating and increasing the sensitivity of the genetic testing process. Notably, other hereditary diseases whose genetic analysis is hampered by similar problems could benefit from this kind of approach. PMID- 11920644 TI - Ovarian cancer antigen CA125 is encoded by the MUC16 mucin gene. AB - Serum assays based on the CA125 antigen are widely used in the monitoring of patients with ovarian cancer; however very little is known about the molecular nature of the CA125 antigen. We recently cloned a partial cDNA (designated MUC16) that codes for a new mucin that is a strong candidate for being the CA125 antigen. This assignment has now been confirmed by transfecting a partial MUC16 cDNA into 2 CA125-negative cell lines and demonstrating the synthesis of CA125 by 3 different assays. Of the 3 antibodies (OC125, M11 and VK-8) tested on the transfected cells, only the first 2 were strongly positive, indicating the differential expression of the CA125 epitopes in these cells. The cloning and expression of CA125 antigen opens the way to an understanding of its function in normal and malignant cells. PMID- 11920645 TI - Monoclonal and polyclonal humoral immune response to EC HER-2/NEU peptides with low similarity to the host's proteome. AB - We are studying peptide immunogenicity as a function of the similarity level to the host's proteome. By using as a model the breast/prostate cancer-associated HER-2/neu antigen, we analyzed the monoclonal and polyclonal humoral immune responses against HER-2/neu peptide motifs not shared with the host proteome. We show here that (i) a mouse monoclonal antibody (MAb) raised against the extracellular domain (EC) of human HER-2/neu oncoprotein recognized a linear peptide motif endowed with low similarity level to the mouse proteome; (ii) likewise, human sera from breast/prostate cancer patients preferentially recognized peptide fragments from the EC of the HER-2/neu oncoprotein having sequences that are not present in the human proteome. Together with previous results obtained in other disease models (cervical cancer-associated HPV16 E7 oncoprotein and Pemphigus vulgaris auto-antigen desmoglein-3), the present data suggest that a low level of sequence similarity to the host's proteome might be an important factor in shaping the pool of B cell epitopes. PMID- 11920646 TI - Phage display particles expressing tumor-specific antigens induce preventive and therapeutic anti-tumor immunity in murine p815 model. AB - The efficacy of phage display particles expressing tumor antigen P1A35-43 in inducing protective and therapeutic anti-tumor immune responses were studied. A protective immune response against a lethal progressive P815 mastocytoma tumor cell challenge was established after subcutaneous injection of phage display particles. Furthermore, the vaccine suppressed growth of pre-existing tumors. Immunization with the hybrid phage particles elicited P1A(35-43) specific CTL responses and a Th1-dominated immune response with phage particle-specific secretion of IFN-gamma but not IL-4. Our results indicate that phage display particles might be a useful vaccine form for tumor-associated antigen epitopes in tumor immunotherapy. PMID- 11920647 TI - Potential markers predicting distant metastasis in axillary node-negative breast carcinoma. AB - Prognostic parameters for axillary node-negative (ANN) breast cancer are still rare. Our aim was to establish potential markers that predict distant metastasis in ANN breast carcinoma and permit detection of the patients with high metastasis risk. A case control study was designed that comprised 64 ANN patients who developed distant metastasis during a 5-10 year follow-up period, 64 ANN patients with recurrence-free survival and 64 node-positive (ANP) comparitors. Immunohistochemistry and/or in situ hybridization were used to detect nm23, Cathepsin-D (Cath-D), Epidermal Growth Factor Receptor (EGFR) and Laminin Receptor (LR) in 192 cases. A significantly lower expression of both nm23 mRNA and protein was found in the ANN-group with poor prognosis compared with the ANN group with good prognosis (p < 0.01). The protein levels of Cath-D, EGFR and LR were significantly higher in the ANN-group with poor prognosis and in the ANP group compared with the ANN-group with good prognosis (p < 0.01 or p < 0.05), but no differences were found between the poor ANN-group and the ANP-group. Multiple regression analysis showed a close correlation of nm23, Cath-D and EGFR expression with occurrence of distant metastasis of ANN breast carcinoma. All markers except nm23 correlated with conventional histopathologic criteria such as tumor grade, margin and vessel invasion. The results suggest the combined detection of nm23, Cath-D and EGFR as predictive markers of distant metastasis in ANN breast cancer patients. Quantitative analysis together with clinicopathologic factors could contribute to estimate the potential risk of metastasis and select individual therapy regimen. PMID- 11920648 TI - Food-derived polyphenols inhibit pancreatic cancer growth through mitochondrial cytochrome C release and apoptosis. AB - There is increasing evidence that food-derived polyphenols have a beneficial effect for cancers. Our purpose was to determine the effect and mechanism of action of these compounds on pancreatic cancer. We measured effects of quercetin on pancreatic cancer in a nude mouse model. We also investigated the effects of quercetin, rutin, trans-resveratrol and genistein on apoptosis and underlying signaling in pancreatic carcinoma cells in vitro. Quercetin decreased primary tumor growth, increased apoptosis and prevented metastasis in a model of pancreatic cancer. In vitro quercetin and trans-resveratrol, but not rutin, markedly enhanced apoptosis, causing mitochondrial depolarization and cytochrome c release followed by caspase-3 activation. In addition, the effect of a combination of quercetin and trans-resveratrol on mitochondrial cytochrome c release and caspase-3 activity was greater than the expected additive response. The inhibition of mitochondrial permeability transition prevented cytochrome c release, caspase-3 activation and apoptosis caused by polyphenols. Nuclear factor kappa B activity was inhibited by quercetin and trans-resveratrol, but not genistein, indicating that this transcription factor is not the only mediator of the polyphenols' effects on apoptosis. The results suggest that food-derived polyphenols inhibit pancreatic cancer growth and prevent metastasis by inducing mitochondrial dysfunction, resulting in cytochrome c release, caspase activation and apoptosis. PMID- 11920649 TI - Physical activity and lung cancer risk in male smokers. AB - We examined the association between physical activity and lung cancer in a prospective cohort of 27,087 male smokers, ages 50-69 years, enrolled in the Alpha-Tocopherol, Beta Carotene Cancer Prevention (ATBC) Study. After an average of 10 years of follow-up, 1,442 lung cancer cases were diagnosed. Cox proportional hazards models were used to estimate the relative risk (RR) and 95% confidence intervals (CI) of lung cancer associated with self-reported occupational and leisure-time activity, adjusted for age, supplement group, body mass index, cigarettes smoked daily, years of smoking, education, energy intake and vegetable intake. There were no associations between occupational, leisure time or combined categories of physical activity with lung cancer risk; however, age appeared to modify the effect of leisure-time activity (p = 0.02). Within increasing quartiles of age, the RRs (CI) for men active in leisure time compared to sedentary men were 0.77 (0.54-1.09), 0.74 (0.57-0.95), 1.09 (0.89-1.33) and 1.03 (0.88-1.21). These data suggest that among smokers, neither occupational nor leisure-time activity is associated with lung cancer risk. There may, however, be some modest risk reduction associated with leisure activity among younger smokers. Published 2002 Wiley-Liss, Inc. PMID- 11920650 TI - Prevalence of germline mutations of MLH1 and MSH2 in hereditary nonpolyposis colorectal cancer families from Spain. AB - HNPCC is an autosomal dominantly inherited cancer-susceptibility syndrome that confers an increased risk for colorectal cancer and endometrial cancer at a young age. It also entails an increased risk of a variety of other tumors, such as ovarian, gastric, uroepithelial and biliary tract cancers. The underlying pathogenic mutation lies in 1 of the 5 known DNA MMR genes (MSH2, MLH1, PMS1, PMS2 and MSH6). We screened a total of 140 individuals from 56 Spanish families with suspected HNPCC for mutations in the DNA mismatch repair genes MLH1 and MSH2, using DGGE and direct DNA sequencing. Families were selected on the basis of a history of HNPCC-related tumors or the occurrence of other associated tumors in members besides the index case affected with colorectal cancer. We detected 14 definite pathogenic germline mutations, 9 in MLH1 and 5 in MSH2 in 13 unrelated families selected by the Amsterdam criteria and Bethesda guidelines (1 family carries 2 mutations) and 3 missense mutations in 3 unrelated families selected by the Amsterdam criteria. Among the 17 germline mutations noted in the Spanish cohort, 10 are novel, 7 in MLH1 and 3 in MSH2, perhaps demonstrating different mutational spectra in the Spanish population, where no founder mutation has been identified. Based on our results, we suggest that in the Spanish population not only HNPCC families fulfilling the Amsterdam criteria but also those following Bethesda guidelines should undergo genetic testing for MSH2 and MLH1 mutations. PMID- 11920651 TI - Reduced poly(ADP-ribosyl)ation in lymphocytes of laryngeal cancer patients: results of a case-control study. AB - Poly(ADP-ribose) polymerase (PARP), a nuclear enzyme that is catalytically activated by DNA strand breaks, plays a complex role in DNA repair. Using NAD(+) as a precursor, it catalyzes the formation of ADP-ribose polymers, which are attached to various proteins. Defects in DNA repair pathways have been associated with increased risks for cancer in humans. We investigated whether differences in the activity of PARP are associated with the risk for laryngeal cancer. In a case control study on genetic, lifestyle and occupational risk factors for laryngeal cancer, PARP activity was assessed as DNA damage-induced poly(ADP-ribose) formation in human peripheral blood lymphocytes by quantitative immunofluorescence analysis. Polymer formation was determined as the cellular response to bleomycin, a well-known inducer of DNA strand breaks, in lymphocytes from 69 laryngeal cancer patients and 125 healthy controls. The frequency of bleomycin-induced polymer formation, measured as mean pixel intensity, was significantly lower in cases (74.6, SE = 3.7) than in controls (94.5, SE = 3.5) and not influenced by smoking, age or sex. There was no significant difference between cases (59.1, SE = 5.2) and controls (50.5, SE = 3.7) in basal polymer formation (in cells not treated with bleomycin). When the highest tertile of polymer formation was used as the reference, the odds ratio for the lowest tertile of bleomycin-induced polymer formation was 3.79 (95% confidence interval 1.37-10.47, p = 0.01). Peripheral blood lymphocytes from laryngeal cancer patients thus showed significantly less bleomycin-induced poly(ADP-ribose) formation. Our results suggest that a reduced capacity of somatic cells to synthesize poly(ADP-ribose) might be associated with an increased risk for laryngeal cancer. The underlying mechanism remains to be investigated. PMID- 11920653 TI - Comparison of aluminium (III) phthalocyanine tetrasulfonate- and meta tetrahydroxyphenylchlorin-monoclonal antibody conjugates for their efficacy in photodynamic therapy in vitro. AB - A challenge in photodynamic therapy (PDT) is to improve the tumour selectivity of the photosensitizers by using monoclonal antibodies (MAbs). With this aim, we developed MAb-conjugates with the hydrophobic photosensitizer meta tetrahydroxyphenylchlorin (mTHPC) and with the hydrophilic sensitizer aluminium (III) phthalocyanine tetrasulfonate (AlPcS(4)). The capacity of these photoimmunoconjugates for selective targeting of squamous cell carcinoma (SCC) in vivo was demonstrated previously in SCC-bearing nude mice. Preliminary in vitro PDT studies with the vulvar SCC cell line A431 showed promising phototoxicity with both sensitizers when coupled to the internalizing MAb 425. To rank the photosensitizers for their potential in photoimmunotherapy, we herein describe an extensive in vitro evaluation of mTHPC-MAb and AlPcS(4)-MAb conjugates. Both classes of conjugates were directly compared using 5 different SCC cell lines as target and 3 different MAbs (BIWA 4, E48 and 425) for tumour cell targeting. In contrast to free AlPcS(4) (IC(50) > or = 700 nM), MAb-conjugated AlPcS(4) was found to be highly phototoxic in PDT in all 5 cell lines. AlPcS(4)-BIWA 4 was most consistently effective with IC(50) values ranging from 0.06-5.4 nM. mTHPC MAb conjugates were in general hardly effective. Phototoxicity (log IC(50)) of the AlPcS(4)-MAb conjugates was found to be strongly correlated with their total cell binding capacity (internalized and surface bound) and to be less correlated with their internalization capacity. In conclusion, these data show a high potential of AlPcS(4)-MAb conjugates in comparison to mTHPC-MAb conjugates for use in PDT. PMID- 11920652 TI - Genetic protection of repopulating hematopoietic cells with an improved MDR1 retrovirus allows administration of intensified chemotherapy following stem cell transplantation in mice. AB - This study was undertaken to analyze the hematotoxicity of paclitaxel (Taxol) and to test whether transduction of repopulating hematopoietic cells with a retroviral vector (SF1m) expressing the human multidrug resistance 1 gene (MDR1) would permit dose intensification following bone marrow transplantation (BMT). While the regimen chosen (8 x 20 mg/kg i.p. within 12 days) produced a non lethal, reversible hematotoxicity in mice with steady-state hematopoiesis, only 35.3% (6/17) of control mice survived when treated starting 14 days post BMT. In contrast, 83.3% (15/18) of mice transplanted with SF1m-transduced cells survived, owing to a significant protection against severe acute myelotoxicity (as determined by neutrophil counts, white and red blood cell counts and values for hemoglobin and hematocrit). After recovery from chemotherapy, an increase of myeloid cells that were resistant to colchicine and effluxed the fluorochrome Rhodamine 123 was observed in SF1m-mice, but not in controls. These results reveal that the lethal, dose-limiting hematotoxicity of an intensified post transplantation chemotherapy with paclitaxel can be prevented by retroviral transfer of the MDR1 gene to a minor proportion of repopulating cells. Our mouse model, mimicking clinically achievable gene transfer rates, thus suggests that bone marrow chemoprotection may widen the therapeutic window and permit an earlier onset of post-transplantation chemotherapy. PMID- 11920654 TI - No evidence for a familial breast cancer susceptibility gene at chromosome 13q21 in Swedish breast cancer families. PMID- 11920656 TI - Osseointegration of hydroxyapatite-coated and noncoated Ti6Al4V implants in the presence of local infection: a comparative histomorphometrical study in rabbits. AB - A study was designed to investigate the osseointegration of titanium implants, either noncoated or coated with hydroxyapatite (HA), into rabbit tibiae in the presence of local infection compared with osseointegration in the absence of local infection. HA-coated or noncoated Ti cylinders were implanted into both tibiae of 32 rabbits (New Zealand Whites). Before implantation the left tibia was contaminated with different quantities of Staphylococcus aureus (10(2)-10(5) CFU). Four weeks after surgery the tibiae were explanted and prepared for microbiological and histomorphometrical examination. Histomorphometrical data, as a representation of implant fixation, were obtained by measuring the percentage of bone around the implants (within a radius of 1 mm from the outer diameter of the implants) and the percentage of the circumference of the implant that was in direct contact with bone. Histomorphometry revealed, in particular for the HA implants, a relationship between the inoculum concentration and/or the presence or absence of infection with the bone contact at the distal implant side. This confirms a relationship between peri-implant infection and bone contact or remodeling. HA-coated implants developed, in the presence of bacteria, more easily a more severe infection than noncoated Ti implants, and we show in the present study that local infection will influence histomorphometrical parameters (bone-implant contact) that determine implant fixation. Precautions to prevent contamination (asepsis) and/or infection (perioperative antibiotics) are even more important for the highly biocompatible HA-coated implant. PMID- 11920657 TI - Platelet adhesion to polystyrene-based surfaces preadsorbed with plasmas selectively depleted in fibrinogen, fibronectin, vitronectin, or von Willebrand's factor. AB - Four plasma proteins have been shown to be able to mediate platelet adhesion to synthetic materials when they are adsorbed as purified proteins: fibrinogen (Fg), fibronectin (Fn), vitronectin (Vn), and von Willebrand factor (vWF). Among them, Fg is thought to play a leading role in mediating platelet adhesion to plasma preadsorbed biomaterials, but this has been established for only three types of materials so far in our laboratory. Furthermore, the role of Fn, Vn, and vWF in mediating platelet adhesion to plasma-preadsorbed surfaces is still unclear. The aim of the current study was to assess the importance of Fg, Fn, Vn, and vWF in mediating platelet adhesion to a series of polystyrene-based surfaces. The strategy applied in the present investigation was to compare platelet adhesion to surfaces preadsorbed with normal plasma, plasma selectively depleted in Fn or Vn or both Fn and Vn, plasma from donors who were genetically deficient in vWF, and serum. Few platelets adhered to the surfaces preadsorbed with serum, whereas depletion of Fn, Vn, or vWF from plasma did not decrease platelet adhesion significantly. Replenishment of exogenous Fg to serum before protein adsorption restored platelet adhesion to the surfaces, suggesting that Fg was the major plasma protein that mediated platelet adhesion. Also, we found that a surface density of adsorbed Fg far below the amount that usually adsorbs to synthetic surfaces was sufficient to support full-scale platelet adhesion. PMID- 11920658 TI - In vitro behavior of albumin-loaded carbonate hydroxyapatite gel. AB - Hydroxyapatite (HA) powder, porous HA, plasma-sprayed HA, apatite cements, and sintered HA have all been investigated as delivery systems for compounds such as human growth hormone and vancomycin. However, many previous studies showed that the period of release was limited to 2-3 weeks. The concept of using a nanoporous matrix as a means of immobilizing proteins is well known but has largely been confined to silica-based systems. Carbonate hydroxyapatite (CHA) is more soluble in vivo than HA, and when formed as an aqueous precipitate, it is often formed as nanocrystals. This study investigated the release profiles of ovine albumin (OVA) from CHA gel stored in phosphate-buffered saline (PBS) and double distilled water (DDW) for times of up to 1 year. It was found that 7.9% OVA could be loaded onto apatitic gels by means of a purely aqueous process. This process provided a simple low-temperature method of protein adsorption on a high surface area apatitic matrix at physiological pH. The rate of short-term release of OVA was lower from CHA gels than from microcrystalline HA powder. However, the period of release from the CHA gel was short term and may have been associated with recrystallization of the gel. OVA loaded into CHA gel was found to remain undegraded in vitro at 37 degrees C for periods of up to 1 year. PMID- 11920659 TI - Effect of elastin on the calcification rate of collagen-elastin matrix systems. AB - The role of elastin in the aortic wall calcification and involved mechanisms were investigated. The major hypothesis of this work is that elastin is one of the major components to regulate calcification of bioprosthetic heart valve (BHV). The relationship between the elastin content and the calcification rate of the aortic wall was established using collagen-elastin matrices (CEM) made of varying ratios of collagen and elastin (90 and 10, 50 and 50, and 20 and 80). Biophysical characterization of CEM was performed by water content measurement and the tensile strength test. The conformational changes of the calcifiable matrix were evaluated as a function of elastin content using Fourier transform infrared (FTIR) spectroscopy. The calcium contents in CEM implanted in the rat subcutaneous model for 7 days were measured using atomic absorption (AA) spectroscopy. As the concentration of elastin in CEM increased from 10 to 80%, the total amount of calcium accumulated in CEM also increased. The calcium level in CEM containing the collagen and an elastin ratio of 20:80 was 20.16 +/- 0.70 microg/mg compared with 1.96 +/- 0.04 microg/mg in CEM containing the collagen and an elastin ratio of 90:10. The calcification rate of CEM pretreated with ethanol increased, as the elastin concentration in the CEM. However, the calcification rate of CEM pretreated with ethanol is significantly lower than that of the untreated control. The permeation rate of ethanol through CEM with the collagen and an elastin ratio of 20:80 (0.37 +/- 0.13 mmol/cm(2)/h) is significantly smaller than that through CEM with 90:10 (0.94 +/- 0.27 mmol/cm(2)/h). These results indicate that elastin has a significant role in tissue calcification and that elastin's resistance to ethanol penetration partially contributes less effectiveness of ethanol on aortic wall calcification. PMID- 11920660 TI - Heparin-like polymers modulate proinflammatory cytokine production by lipopolysaccharide-stimulated human monocytes. AB - The search for heparin-like materials remains an intensive field of research. In this context, we studied the immunomodulatory properties of semisynthetic dextran derivatives and naturally occurring sulfated polysaccharides present in brown seaweed (fucans). In this study, we investigated the functional potencies of fucan and dextran derivatives by analyzing their effects on the release of proinflammatory cytokines by resting or lipopolysaccharide (LPS)-stimulated human monocytes and their interactions on monocyte surfaces. The results showed that fucan, dextran derivatives, and heparin differentially (1) triggered interleukin 1alpha, tumor necrosis factor alpha, interleukin-6, and interleukin-8 production by monocytes in a dose-dependent manner, (2) modulated cytokine production by LPS stimulated monocytes, and (3) specifically inhibited the binding of biotinylated LPS to monocyte membranes. Taken together, these data indicated that fucan and dextran derivatives displayed interesting immunomodulatory effects on human blood cells that could be relevant as new drugs or biomaterial coatings. Indeed, such polysaccharides, by regulating monocyte activation, could contribute to the improved biocompatibility of implants. PMID- 11920661 TI - Synergistic effects of glucose and ultraviolet irradiation on the physical properties of collagen. AB - Our objective was to strengthen and stabilize collagen films without the introduction of cytotoxic chemical crosslinkers. We hypothesized that collagen could be rapidly crosslinked with glucose with ultraviolet (UV) irradiation as a catalyst. In theory, UV-generated free radicals can expedite collagen crosslinking with glucose via the formation of reactive, linear glucose molecules. The mechanical properties of glucose-incorporated, UV-exposed collagen films and appropriate controls were determined under various conditions: (1) hydration in phosphate-buffered saline, (2) heat-denaturation, (3) incubation in a collagenase solution, and (4) incubation in a trypsin solution. Without exposure to UV, the incorporation of glucose into the films had no effect. Without glucose, exposure to UV increased the strength but caused significant denaturation. The combination of glucose and UV, however, synergistically improved the mechanical properties and enzyme resistance of collagen films, indicative of increased crosslinking without significant denaturation effects. The addition of thiourea, a potent free-radical scavenger, or aminoguanidine, an inhibitor of glucose-derived crosslinking, to the collagen films markedly hindered these synergistic effects. These data strongly suggest that free-radical dependent, glucose-derived crosslinks provide the enhanced strength and enzyme resistance observed in glucose-incorporated, UV-exposed collagen films. Further studies are required to explore biomaterial applications of this novel collagen crosslinking method. PMID- 11920662 TI - TGF-beta, BMPS, and their signal transducing mediators, Smads, in rat fracture healing. AB - Smads are cytoplasmic signal transducers of transforming growth factor-beta (TGF beta) and bone morphogenetic proteins (BMPs). Their relation to fracture healing is unknown. This study examined the temporal protein expression of Smads, together with TGF-beta and BMPs, using immunohistochemistry in a rodent fracture model. Over-expression of TGF-beta, BMPs-2, 4, and 7, common-mediator Smad (Smad4), and receptor-regulated Smads (Smads1, 2, 3, and 5) versus lower levels of inhibitory Smad (Smad6), were detected at day 3 in osteogenic cells in the thickened periosteum and bone marrow at the fracture sites. At day 10, Smad6 increased dramatically, Smad2, Smad3, and Smad4 remained elevated while Smad1 and Smad5 decreased in the fracture callus. Smad7 was expressed only in vascular endothelial cells. By day 28, when new bone had replaced the fracture callus, all the protein regulators decreased, approaching control levels. During fracture healing, the expression patterns of Smads1 and 5 were similar to that of BMPs-2 and 7 whereas the expression of Smads2 and 3 was parallel with that of TGF-beta. The Smad family, associated with BMPs and TGF-beta, may play an important role in the early stage of rat fracture healing. PMID- 11920663 TI - Surface modification of poly (D,L-lactic acid) with chitosan and its effects on the culture of osteoblasts in vitro. AB - Chitosan is a good biodegradable natural polymer, widely used in biomedical fields. In this study, chitosan was used to modify the surface of poly (D,L lactic acid) (PDLLA) in order to enhance its cell affinity. The properties of a modified PDLLA surface and control were investigated by contact angle and electron spectroscopy for chemical analysis (ESCA), which indicated the changes in surface energy and chemical structure. Scanning electron microscopy (SEM) observation displayed differences in surface morphology between the chitosan modified film and the control. These data reflected that PDLLA films could be modified with chitosan and in turn may affect the biocompatibility of the modified films. Therefore, adhesion and growth of osteoblasts on modified PDLLA films as well as control were studied. Cell morphologies on the films were examined by SEM and cell viability was evaluated using an MTT assay; the differentiated cell function was assessed by measuring alkaline phosphatase (ALP) activity. The ALP activity of modified PDLLA films was significantly higher than that found on the control (p < 0.01). The proliferation of osteoblasts on modified films was also found to be higher than that on the control (p < 0.05), suggesting that chitosan could be used to modify PDLLA and then enhance its cell biocompatibility. PMID- 11920664 TI - Cationic lipid-mediated transfection of bovine aortic endothelial cells inhibits their attachment. AB - The need for a small-diameter vascular graft for coronary artery and peripheral vascular replacement is great and is projected to increase as the population ages. Synthetic small-diameter vascular grafts fail because of acute thrombosis or chronic intimal hyperplasia leading to restenosis. Endothelial cell seeding has been attempted with limited success in the femoral artery by Zilla and others. However, patency rates have not increased sufficiently to justify large clinical trials. Genetic engineering of endothelial cells before seeding has been proposed to encourage endothelial cell phenotypes that would predispose the graft to patency. In this study, we investigate the effect cationic lipid-mediated transfection of endothelial cells with respect to their attachment to a potential graft material, Fluoropassiv (Vascutek). Liposomal transfection was optimized for maximum gene expression. We report that transfection decreases the ability of bovine aortic endothelial cells to attach by approximately 100% as compared with nontransfected control over 18 h. Further, when placed under physiologic shear conditions, this difference is sustained. The effects of gene transfer on endothelial cell adhesion must be included as an important optimization criterion along with gene expression for engineered endothelial cell-seeding applications. PMID- 11920665 TI - Polyethylene wear characteristics in vivo and in a knee stimulator. AB - Validation of a wear simulator requires that the device produce a similar type and amount of wear and particles of a comparable morphology as occurs clinically. Using techniques previously established to compare polyethylene particles from hip simulators to those from retrieved tissues, particles isolated from six revised posterior stabilized knee replacements were characterized and compared to particles generated from the same knee design worn in a knee simulator. The particles produced in the knee simulator were of comparable size but had less variability in their form factor compared to the particles produced in vivo. Comparable wear features were seen on the articulating surfaces in both groups. These results indicate that this knee joint simulator is able to reproduce a baseline type of wear that is similar to that in vivo and should encourage further use of this device to better understand knee component wear and function. PMID- 11920666 TI - Concentration- and composition-dependent effects of metal ions on human MG-63 osteoblasts. AB - Metal debris from implants has been shown to alter the function of osteoblasts in cell cultures. Its remains unclear, however, if specific forms of released ionic metals are involved in the pathogenesis of periprosthetic osteolysis. We evaluated the relative effects of ionic forms of implant metals by treating human osteoblast-like MG-63 osteosarcoma cells with eight concentrations (0.001-10.0 mM) of Cr(+3), Mo(+5), Al(+3), Ta(+5), Co(+2), Ni(+2), Fe(+3), Cu(+2), Mn(+2), Mg(+2), Na(+2), and V(+3) chloride solutions. The results demonstrated that the metal ions differentially affected osteoblast proliferation, viability, type-I collagen gene expression, and cytokine release. The metal ions were ranked in order from least to most toxic (based on a 50% reduction in viability) as follows: Na < Cr < Mg < Mo < Al < Ta < Co < Ni < Fe < Cu < Mn < V. Metal-induced decreases in osteoblast proliferation were similar in ranking. Nontoxic concentrations of metals had no effect on procollagen alpha1[I] gene expression; only at toxic concentrations did metals produce a decrease in gene expression. The most toxic metals (V, Mn, Fe, and Ni) were also the only metals found to induce IL-6 secretion on a per cell basis (of the cytokines tested, interleukin 6 (IL-6), interleukin beta 1 (IL-1beta), transforming growth factor beta 1 (TGF beta1), and tumor necrosis factor alpha (TNF-alpha), only IL-6 was detectable in the culture medium after 48 h for any metal at any concentration). Less toxic metals (e.g., Co and Cr) had little effect on IL-6 release, even at high concentrations. In general, metal ions reduced osteoblast function (i.e., proliferation and collagen gene expression) in proportion to the degree of toxicity. These results support the hypothesis that adverse local cellular responses (particularly necrotic responses) associated with metal debris from implanted metallic devices may be due in part to metal ions released from implants or from particulate debris. PMID- 11920667 TI - Use of Ti-coated replicas to investigate the effects on fibroblast shape of surfaces with varying roughness and constant chemical composition. AB - A two-stage replica technique with a subsequent titanium (Ti)-coating treatment was used to faithfully replicate topographies of polished, acid-etched, machined like, finely blasted, coarsely blasted, coarsely blasted and acid-etched, and Ti plasma-sprayed Ti surfaces. The replicas were used to study the influence of different rough surface topographies on the response of human fibroblasts in vitro under conditions of constant surface chemistry for all surfaces. The surface topographies of the replicas were characterized using non-contact laser profilometry, scanning electron microscopy (SEM), and stereo-SEM, whereas surface chemistry was examined using X-ray photoelectron spectroscopy. Fibroblasts were trypsinized and plated onto the Ti-coated epoxy-resin replica surfaces for 24 h and observed with SEM. Fluorescein-5-thiosemicarbazide was used to stain the cell components including cell membrane, and the stained cells were optically sectioned using epifluorescent microscopy. The optical sections were computationally reconstructed to obtain three-dimensional images and cell volume and cell thickness determined. The different surface topographies were found to alter cell thickness and cell morphology. However, cell volume as computed from three-dimensional reconstructions was not affected by surface features. The results suggest that cells distort themselves to accommodate to rough surfaces but their volume is not significantly altered. PMID- 11920668 TI - Transforming growth factor-beta 1, 2, and 3 can inhibit epithelial tissue outgrowth on smooth and microgrooved substrates. AB - In this study, we describe the influence of parallel surface microgrooves, and of TGF-beta, on the outgrowth of corneal epithelial tissue. Microgrooves (depth 1 microm, width 1-10 microm) were made in polystyrene culturing surfaces. These surfaces were left untreated, or loaded with TGF-beta 1, 2, or 3 (6.0 ng/cm(2)). Subsequently, epithelial explants from bovine corneas were placed on the experimental surfaces. After 9 days of culturing, tissue outgrowth was evaluated. Furthermore, the tissue cultures were analyzed histologically. It was shown that epithelial tissue grew from the explants over all experimental surfaces. On microgrooved surfaces outgrowth proceeded in the direction of the grooves, rather than perpendicular to the grooves. The addition of each type of TGF-beta resulted in a reduction of outgrowth. However, outgrowth remained directed by the grooves. Further, the explants had shrunk after TGF treatment. Histology showed that this shrinkage was not related to alpha-smooth muscle actin expression in the explants. We conclude that microgrooves can direct, and TGF-betas can inhibit the outgrowth of epithelial tissue. This finding could be useful in biomaterial applications where the growth of epithelial tissue needs to be discouraged. PMID- 11920669 TI - Enzymatic responses of human deciduous pulpal fibroblasts to dental restorative materials. AB - The purpose of this study was to evaluate the responses of succinic dehydrogenase (SDH) and alkaline phosphatase (ALP) activities of human deciduous teeth pulpal fibroblasts (HDPF) to dental restorative materials. Tested materials included Z100 (3M), Dyract (Dentsply), FujiII (GC), and FujiIILC (GC). IRM (Dentsply) and culture medium (MD) alone were used as positive and negative controls, respectively. Specimens 6 mm (diameter) x 3 mm were prepared in accordance with manufacturers' instructions. For light-cured materials, specimens were light cured for 40 s on both sides under a celluloid strip. For chemical-cured materials, specimens were allowed to set at room temperature for 15 min. The specimens were immersed in 1 mL of culture medium without serum for 24 h at room temperature. The extracts were filtered through 0.22-mm filters. HDPF (10,000 cells/well) was incubated with 100 microL of extract and 20 % FBS in a 96-well plate for 24 h in a 37 degrees, 5 % CO(2) incubator. Six wells per material were prepared. Optical density (OD) of SDH and ALP of HDPF were measured by a spectrophotometer. The means were analyzed by ANOVA and then a Duncan Test. The ranking of OD of SDH was IRM < FujiIILC < FujiII = Z100 < Dyract < MD (p < 0.05). The ranking of OD of ALP was IRM < Z100 = Dyract < FujiII < FujiIILC < MD (p < 0.05). The result showed that all of the tested restorative materials were cytotoxic to human deciduous pulpal fibroblasts. The cytotoxicity of resin modified glass ionomer cements (FujiIILC) was stronger than that of traditional glass ionomer cements (FujiII) and composite resin (Z100), and that of compomer (Dyract) was the weakest. On the contrary, ALP activities of resin-modified glass ionomer cements (FujiIILC) and composite resin (Z100) were higher than those of traditional glass ionomer cements (FujiII), while those of compomer (Dyract) were the lowest. It is concluded that, in this study, FujiIILC was the most cytotoxic material and the least inhibitive of ALP activities, Dyract was the weakest cytotoxic material and had the highest inhibition of ALP activities. The rankings of the MTT assay and the ALP assay were not consistent. PMID- 11920670 TI - Effect of chemical composition on the corrosion behavior of Ni-Cr-Mo dental casting alloys. AB - The objective of this investigation was to study the compositional influence on the corrosion behavior of Ni-Cr-Mo dental casting alloys in acidic artificial saliva. Cyclic potentiodynamic and potentiostatic tests were used to evaluate the corrosion behavior of different Ni-Cr-Mo dental casting alloys in deaerated artificial saliva with pH 5 at 37 degrees C. Optical microscope observations were made following the cyclic potentiodynamic tests. Surface chemical analyses were characterized by X-ray photoelectron spectroscopy and auger electron spectroscopy following the potentiostatic tests. The results show that the corrosion resistance of the Ni-Cr-Mo casting alloys investigated is associated with the formation of passive film containing Ni(OH)(2), NiO, Cr(2)O(3), and MoO(3), on the surface. The pitting potential and passive range, respectively, were statistically different among the different Ni-Cr-Mo alloys. The Ni-Cr-Mo alloys with higher Cr ( approximately 21%) and Mo ( approximately 8%) contents had a much larger passive range in the polarization curve and were immune to pitting corrosion due to the presence of high Cr (maximum approximately 31-35%) and Mo (maximum approximately 12%) contents in the surface passive film. The presence of Ti lower than 4% in the Ni-Cr-Mo casting alloy had no effect on corrosion resistance. A pitting resistance equivalent (PRE) of about 49 could provide the Ni-Cr-Mo alloy with a good pitting corrosion resistance. PMID- 11920671 TI - Novel osteoblast-adhesive peptides for dental/orthopedic biomaterials. AB - Next generation dental/orthopedic biomaterials must be designed to enhance and support osteoblast adhesion. The osteoblasts use different ways to adhere, that is, integrin- and proteoglycan-mediated mechanisms. The present study reports on the synthesis and osteoblast-adhesive properties of peptides carrying RGD motifs and of sequences mapped on human vitronectin. Our data suggest that osteoblast adhesion on polystyrene plates modified with a linear peptide, in which the GRGDSP sequence is repeated four times, was significantly higher when compared to the adhesion obtained using branched peptides, interestingly containing the same motif. Osteoblast adhesion assays on acellular bone matrix using this active peptide gave very promising results. We also demonstrated that a novel peptide, carrying the X-B-B-B-X-B-B-X motif (where B is a basic amino acid and X is a nonbasic residue), promotes proteoglycan-mediated osteoblast adhesion more efficiently with respect to the KRSR sequence that was recently proposed as heparan-sulfate binding peptide. PMID- 11920672 TI - In vitro generation of differentiated cardiac myofibers on micropatterned laminin surfaces. AB - Cardiac muscle fibers consist of highly aligned cardiomyocytes containing myofibrils oriented parallel to the fiber axis, and successive cardiomyocytes are interconnected at their ends through specialized junctional complexes (intercalated disks). Cell culture studies of cardiac myofibrils and intercalated disks are complicated by the fact that cardiomyocytes become extremely flattened and exhibit disorganized myofibrils and diffuse intercellular junctions with neighboring cells. In this study we sought to direct the organization of cultured cardiomyocytes to more closely resemble that found in vivo. Lanes of laminin 5-50 microm wide were microcontact-printed onto nonadhesive (BSA-coated) surfaces. Adherent cardiomyocytes responded to the spatial constraints by forming elongated, rod-shaped cells whose myofibrils aligned parallel to the laminin lanes. Patterned cardiomyocytes displayed a striking, bipolar localization of the junction molecules N-cadherin and connexin43 that ultrastructurally resembled intercalated disks. When laminin lanes were widely spaced, each lane of cardiomyocytes beat independently, but with narrow-spacing cells bridged between lanes, yielding aligned fields of synchronously beating cardiomyocytes. Similar cardiomyocyte patterns were achieved on the biodegradable polymer PLGA, suggesting that patterned cardiomyocytes could be used in myocardial tissue engineering. Such highly patterned cultures could be used in cell biology and physiology studies, which require accurate reproduction of native myocardial architecture. PMID- 11920673 TI - Uptake of poly(D,L-lactic-co-glycolic acid) microspheres by antigen-presenting cells in vivo. AB - Dendritic cells are the most potent antigen-presenting cells (APC) and the most effective stimulators of primary T cell responses. Based on the strong influence of the APC on the immune response, we investigated cellular uptake of a biodegradable antigen delivery system, poly(D,L-lactic-co-glycolic acid) (PLGA) microspheres, at two sites of injection: intraperitoneal and intradermal. We hypothesized that a fluorescent probe, tetramethylrhodamine labeled dextran, loaded in PLGA microspheres would be taken up by APCs and thereby provide a means for studying cellular uptake of PLGA microspheres in vivo. Phagocytic load and cell phenotype were determined using flow cytometry and confocal laser scanning microscopy. The results revealed cellular uptake of tetramethylrhodamine dextran loaded PLGA microspheres at both injection sites. After intraperitoneal immunization, the predominant cell phagocytosing PLGA microspheres in the peritoneal cavity was the macrophage whereas the intradermal immunization resulted in uptake of PLGA microspheres by dendritic cells. Hence, these results suggest that the profile for cellular uptake varies with the site of injection. More importantly, this study provides direct and conclusive evidence of uptake of PLGA microspheres by the most potent APC, the dendritic cell. PMID- 11920674 TI - Protein and surface effects on monocyte and macrophage adhesion, maturation, and survival. AB - Cell adhesion and maturation can be affected by the protein adsorption profile on the surface of an implanted biomaterial. In this study we have investigated how surface chemistry and adsorbed proteins can modulate monocyte and macrophage adhesion, IL-13-induced foreign-body giant cell formation, and apoptosis in vitro. Compared to a dimethylsilane-modified surface (DM), a surface modified with RGD peptides had no effect on adhesion density, foreign-body giant cell (FBGC) formation, or apoptosis in nondepleted serum conditions. The depletion of specific adhesive proteins affected adhesion, FBGC formation, and apo- ptosis. While the depletion of fibronectin and vitronectin had no overall effect compared to nondepleted serum conditions, the depletion of IgG from serum caused a significant decrease in initial adherent cell density [1000 +/- 200 compared to 2460 +/- 590 (p = 0.02)], a significant decrease in FBGC formation [2% compared to 17% (p = 0.02)], and a significant increase in the level of apoptosis [57% compared to 32% (p = 0.01)] on DM. The lowered initial adherent cell density on DM was not observed on the RGD surface, indicating that the RGD surface promotes increased initial adhesion. However, the RGD surface does not affect FBGC formation (i.e., macrophage fusion) or levels of apoptosis, which remained comparable to those on the DM surfaces at days 7 and 10. PMID- 11920675 TI - Heparin-like polymer improved healing of gastric and colic ulceration. AB - A family of chemically substituted biopolymers has been developed to protect and stabilize heparin binding growth factors and was shown to enhance tissue repair in various in vivo models. One of these compounds, a dextran derivative named RGTA11, was tested for its ability to treat acute gastritis and colic ulceration models induced by ethanol and acid. RGTA was not efficient in reducing nor in protecting against gastric acidic secretion compared to EGF. Ethanol gastritis measured by the alteration score of the injured mucosa was reduced by 56% with the oral administration of RGTA at doses of 100 microg/kg (p < 0.01). A similar effect was obtained by PGE2 at a similar dose. Alterations of the colic mucosa were reduced after 72 h by 75% after oral administration of RGTA11. RGTA presents both anti-inflammatory and tissue repair activities mediated by growth factor protection. These two properties would be beneficial for digestive ulcer treatment. The results presented here provide evidence for these effects. PMID- 11920676 TI - Biocompatibility evaluation of ePTFE membrane modified with PEG in atmospheric pressure glow discharge. AB - ePTFE membranes were modified by poly(ethylene glycol) having a molecular weight of 600 (PEG-600) in atmospheric pressure glow discharge (APG) plasma treatment. ePTFE membranes were immersed in 1%, 3%, or 5% (w/v) PEG-600 in dehydrated ethanol. PEG-600-penetrated ePTFE membranes were dried in a vacuum to immediately remove ethanol, then treated with APG at 20 kHz and 60-70 W for 15 min and thoroughly washed with ethanol and water. PEG-600-modified ePTFE membranes were analyzed using contact angle measurement, Fourier transform infrared attenuated total reflectance (FTIR-ATR), and scanning electron microscopy (SEM). ePTFE membrane contact angles were reduced after PEG-600 plasma treatment. FTIR-ATR spectra showed an absorption band due to a PEG hydroxyl group (-OH). SEM showed that ePTFE fiber surfaces were uniformly immobilized with PEG-600 and retained their porous structure. A general biological evaluation of the PEG-modified ePTFE membranes showed no cytotoxicity on CHO-K1 cell lines and no hemolytic action. Albumin adsorption on the PEG-modified ePTFE membranes increased with increasing PEG-600 deposited on ePTFE membranes. Fibrinogen adsorption decreased with increasing PEG-600 deposited on ePTFE membranes. gamma-Globulin adsorption did not change before or after PEG plasma modification. 1% and 3% PEG-600 plasma treated ePTFE only slightly increased platelet adhesion, but adhering platelets evidenced no pseudopod formation. 5% PEG-600-modified ePTFE showed relatively large numbers of platelet adhesion. We concluded that 3% PEG-600-modified ePTFE membrane had the best physical properties and biological compatibility, indicating 3% PEG-600-modified ePTFE membranes exhibit the potential for blood filter application. PMID- 11920677 TI - Regulation of cell proliferation by Smad proteins. AB - Transforming growth factor-beta (TGF-beta) family members which include TGF betas, activins, and bone morphogenetic proteins (BMPs) regulate a broad spectrum of biological responses on a large variety of cell types. TGF-beta family members initiate their cellular responses by binding to distinct receptors with intrinsic serine/threonine kinase activity and activation of specific downstream intracellular effectors termed Smad proteins. Smads relay the signal from the cell membrane to the nucleus, where they affect the transcription of target genes. Smad activation, subcellular distribution, and stability have been found to be intricately regulated and a broad array of transcription factors have been identified as Smad partners. Important activities of TGF-beta are its potent anti mitogenic and pro-apoptotic effects that, at least in part, are mediated via Smad proteins. Escape from TGF-beta/Smad-induced growth inhibition and apoptosis is frequently observed in tumors. Certain Smads have been found to be mutated in specific types of cancer and gene ablation of particular Smads in mice has revealed increased rate of tumorigenesis. In late stage tumors, TGF-beta has been shown to function as a tumor promoter. TGF-beta can stimulate the de differentiation of epithelial cells to malignant invasive and metastatic fibroblastic cells. Interestingly, TGF-beta may mediate these effects directly on tumor cells via subverted Smad-dependent and/or Smad-independent pathways. PMID- 11920678 TI - D1 dopamine receptor: a putative neurochemical and behavioral link to cocaine action. AB - Cocaine is one of the most abused psychostimulants known to man and as such, researchers have been steadfast in their attempts to understand the neurobiological mechanisms responsible for its abuse. Cocaine undoubtedly wreaks havoc on a number of mammalian neuronal neurotransmitter systems, and it is maintained that this dysregulatory effect supports cocaine abuse. Cocaine's mechanism of action has been well described. Cocaine binds differentially to the dopamine, serotonin, and norepinephrine transport proteins and directly prevents the re-uptake of dopamine, serotonin, and norepinephrine into pre-synaptic neurons (Heikkila et al., 1975, Biochem Pharmacol 24(8):847-852; Reith et al., 1986, Biochem Pharmacol 35(7):1123-1129; Ritz et al., 1987, Science 237:1219 1223). Inhibition of re-uptake subsequently elevates the synaptic concentrations of each of these neurotransmitters. In addition to this direct effect, cocaine also produces a number of indirect actions, which alter other neuromodulatory systems (i.e., opioidergic, glutamatergic, and GABAergic systems). Many of these effects are just beginning to be elucidated, but nonetheless contribute to this agent's diverse pharmacological profile. Interestingly, it is the indirect actions of this mellifluous molecule, which mediate most of its sought and unsought effects. The intricacy with which cocaine produces neuronal alterations beyond its direct effects on neurotransmitter re-uptake appear to be most relevant to cocaine abuse, and hence the phenomenon of addiction. In light of cocaine's multifarious effects on numerous neuronal systems, its effect on dopaminergic neurotransmission has attracted the most attention, particularly because of the implicated role of dopamine in brain reward. Pharmacologically, the psychostimulant effects of cocaine appear to be mediated by its ability to enhance dopaminergic activity within the mesocorticolimbic circuit (Roberts et al., 1977, Pharmacol Biochem Behav 6(6):615-620). Additionally, it is the intensity with which cocaine produces alterations in dopaminergic circuitry that have enabled this drug to prevail as one of the most addictive substances known to man. This review will summarize findings relevant to cocaine-induced alterations in dopamine-mediated signal transduction. Specifically, it will concentrate on the D1 dopamine receptor and intracellular signaling mediated by this receptor subtype. It will describe cocaine-induced cellular and behavioral alterations relevant to this pathway and how these changes potentially effect gene transcription and protein expression. This article too will review a common behavioral manifestation associated with repeated cocaine exposure, sensitization, and why the D1 dopamine receptor and its associated signaling pathway have been implicated in this phenomenon. Lastly, this article will discuss how targeting the D1 dopamine receptor and its signaling pathway may offer some insight into understanding cocaine addiction, a somewhat elusive brain disease. PMID- 11920679 TI - DNA mismatch repair and mutation avoidance pathways. AB - Unpaired and mispaired bases in DNA can arise by replication errors, spontaneous or induced base modifications, and during recombination. The major pathway for correction of mismatches arising during replication is the MutHLS pathway of Escherichia coli and related pathways in other organisms. MutS initiates repair by binding to the mismatch, and activates together with MutL the MutH endonuclease, which incises at hemimethylated dam sites and thereby mediates strand discrimination. Multiple MutS and MutL homologues exist in eukaryotes, which play different roles in the mismatch repair (MMR) pathway or in recombination. No MutH homologues have been identified in eukaryotes, suggesting that strand discrimination is different to E. coli. Repair can be initiated by the heterodimers MSH2-MSH6 (MutSalpha) and MSH2-MSH3 (MutSbeta). Interestingly, MSH3 (and thus MutSbeta) is missing in some genomes, as for example in Drosophila, or is present as in Schizosaccharomyces pombe but appears to play no role in MMR. MLH1-PMS1 (MutLalpha) is the major MutL homologous heterodimer. Again some, but not all, eukaryotes have additional MutL homologues, which all form a heterodimer with MLH1 and which play a minor role in MMR. Additional factors with a possible function in eukaryotic MMR are PCNA, EXO1, and the DNA polymerases delta and epsilon. MMR-independent pathways or factors that can process some types of mismatches in DNA are nucleotide-excision repair (NER), some base excision repair (BER) glycosylases, and the flap endonuclease FEN-1. A pathway has been identified in Saccharomyces cerevisiae and human that corrects loops with about 16 to several hundreds of unpaired nucleotides. Such large loops cannot be processed by MMR. PMID- 11920680 TI - Low oxygen tension stimulates collagen synthesis and COL1A1 transcription through the action of TGF-beta1. AB - Recent findings point to low oxygen tension (hypoxia) as an important mechanism for the expression of several eukaryotic genes. We have previously shown that hypoxia (2% O2), when compared to standard oxygen tension (20% O2), upregulates the mRNA levels of the human alpha1(I) (COL1A1) procollagen gene and transforming growth factor-beta1 (TGF-beta1) in human dermal fibroblasts. In this report, we determined the effect of hypoxia on collagen synthesis and transcription. Exposure of human dermal fibroblasts to hypoxia for 24-72 h led to a threefold, dose-dependent increase in collagenous protein (P < 0.0001; r = 0.9794) and to enhanced type I procollagen deposition, as shown by direct immunofluorescence. Transient transfections with a series of luciferase- and CAT-promoter constructs of the human COL1A1 gene (spanning from -2.5 kb to +113 bp) showed that hypoxia increases the transcriptional activity of constructs having 5' endpoints between 804 bp and -107 bp, with loss of stimulation at -84 bp. Maximal increase in promoter activity in hypoxia was observed between -190 and -174 bp of the proximal promoter, once a cKrox repressor site (-199 to -224 bp) was deleted. Upregulation of COL1A1 mRNA levels in hypoxia was blocked by a TGF-beta1 anti sense oligonucleotide, and failed to occur in fibroblasts from TGF-beta1 knock out mice. Co-transfection and overexpression with a Smad7 construct abrogated the increase in COL1A1 promoter activity observed in hypoxia. Upregulated transcriptional activity of the TGF-beta1 promoter in hypoxia was found to be maximal between -453 and -175 bp from the transcriptional start site. Since hypoxia is a critical feature of the early phases of wound repair, we conclude that it may act as a potent physiologic stimulus for collagen synthesis. TGF beta1 appears to be a critical component of this response. PMID- 11920681 TI - Effects of mast cells on the behavior of isolated heart fibroblasts: modulation of collagen remodeling and gene expression. AB - The extracellular matrix plays a critical role in the development and maintenance of the vertebrate heart. Changes in the accumulation, composition, or organization of the extracellular matrix are known to deleteriously affect heart function. Mast cells are thought to stimulate collagen expression and fibroblast proliferation accompanying fibrosis in some organs; however, the effects of mast cells on the heart interstitium are largely unexplored. The present studies were carried out to determine the effects of mast cells on isolated heart fibroblasts. Several in vitro assays were used including collagen gel contraction to examine the effects of mast cells on the function of isolated fibroblasts. Neonatal heart fibroblasts were cultured either with mast cells, mast cell-conditioned medium, or mast cell extracts, and their ability to contract collagen gels measured. Results from these experiments indicated that mast cells inhibit heart fibroblast migration and contraction of 3-dimensional collagen gels. Further experiments indicated that incubation of neonatal heart fibroblasts with extracts of mast cells altered the expression of collagen, matrix metalloproteases, and matrix receptors of the integrin family. These studies suggest that mast cells play an important role in the regulation of the cardiac interstitial matrix. Further studies are warranted to determine the mechanisms whereby mast cells modulate fibroblast activity. PMID- 11920682 TI - Expression and targeting of the tight junction protein CLDN1 in CLDN1-negative human breast tumor cells. AB - Claudins and occludin constitute the major transmembrane proteins of tight junctions (TJs). We have previously identified the human homologue of the murine Cldn1, CLDN1 (SEMP1) that is expressed in normal, mammary gland-derived epithelial cells but is absent in most human breast cancer cell lines. To investigate the potential functions of CLDN1 protein in tumor and normal epithelial cells, we developed an I-NGFR retroviral vector and monoclonal anti CLDN1 antibody. In subconfluent and confluent breast cancer cells, MDA-MB-435 and MDA-MB-361, endogenous CLDN1 expression was not detected by an anti-CLDN1 monoclonal antibody by Western blot analysis or quantitative RT-PCR. When CLDN1 negative breast cancer cell lines were transduced with a CLDN1 retrovirus the cells express CLDN1 mRNA constitutively as shown by quantitative RT-PCR. Immunofluorescence analyses of the CLDN1 retroviral transduced breast tumor cells using monoclonal antibodies against CLDN1 reveals a subcellular distribution at cell-cell contact sites similar to the CLDN1 homing pattern in T47-D cells, which express endogenous CLDN1. This cell-cell contact co-localization of CLDN1 was evident in CLDN1-transduced breast tumor cells which fail to express occludin protein (MDA-MB-361 and MDA-MB-435) and express relatively little ZO-1 protein (MDA-MB-435), suggesting that other proteins may be responsible for targeting of CLDN1 to cell-cell contact sites. The re-expression of CLDN1 decreases the paracellular flux of 3 and 40 kDa dextran despite the absence of occludin in the MDA-MB-361 tumor cells. Our findings indicate that in CLDN1-negative breast tumor cells, the basal protein partner requirements for physiological homing of the CLDN1 protein are intact, and that CLDN1 gene transfer and protein expression itself might be sufficient to exert a TJ-mediate gate function in metastatic tumor cells even in the absence of other TJ-associated proteins, such as occludin. PMID- 11920683 TI - Merosin-integrin promotion of skeletal myofiber cell survival: Differentiation state-distinct involvement of p60Fyn tyrosine kinase and p38alpha stress activated MAP kinase. AB - Myofiber survival and suppression of anoikis depend in large part on the merosin (laminin-2/-4)-integrin alpha7beta1D cell adhesion system; however, the question remains as to the nature of the signaling molecules/pathways involved. In the present study, we investigated this question using the C2C12 cell model of myogenic differentiation and its merosin- and laminin-deficient derivatives. Herein, we report that: 1) of four members of the Src family of tyrosine kinases studied (p60Src, p53/56Lyn, p59Yes, or p60Fyn), the expression and activity of p60Fyn are found in myotubes exclusively; 2) a severe decrease of p60Fyn activity correlates with myotube apoptosis/anoikis induced by pharmocological compounds (herbimycin A or PP2) which inhibit tyrosine kinases of the Src family, by merosin deficiency and by beta1 integrin inhibition; 3) myoblast survival depends on Fak and the MEK/Erk pathway, in contrast to myotubes; 4) the PI3-K pathway is not involved in either myoblast or myotube survival; and 5) p38alpha SAPK stimulation and activity (but not that of p38beta) are required in the progression of myotube apoptosis/anoikis induced by p60Fyn inhibition, merosin deficiency or beta1 integrin-inhibition; however, p38 is not involved in myoblast apoptosis. Taken together, these results suggest that the promotion of myotube survival by the merosin-alpha7beta1D adhesion system involves p60Fyn, and that disruptions in this cell adhesion system induce myotube apoptosis/anoikis through a p38alpha SAPK-dependent pathway. PMID- 11920684 TI - Epidermal growth factor stimulates serine/threonine phosphorylation of the focal adhesion protein paxillin in a MEK-dependent manner in normal rat kidney cells. AB - Epidermal growth factor (EGF)-stimulated proliferation of renal epithelial cells plays an important role in the recovery of kidney tubule epithelia following exposure to insult. Numerous studies have demonstrated that tyrosine phosphorylation of the focal adhesion protein paxillin mediates in part the effects of growth factors on cell growth, migration, and organization of the actin-based cytoskeleton. The experiments in this report were designed to determine the effect of EGF on paxillin phosphorylation in normal rat kidney (NRK) epithelial cells. Interestingly, treatment of NRK cells with EGF stimulated paxillin serine/threonine phosphorylation, which caused a reduction in the mobility of paxillin on sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE). The EGF-stimulated mobility shift of paxillin was independent of an intact cytoskeleton, phosphatidylinositol 3-kinase (PI 3-kinase) activation, protein kinase C (PKC) activation, and cellular adhesion. However, inhibitors of the mitogen-activated protein kinase/extracellular signal-regulated kinase kinase abrogated the EGF-stimulated change in paxillin mobility. In addition, the EGF stimulated change in paxillin serine/threonine phosphorylation was not accompanied by a profound reorganization of the actin cytoskeleton. These results identify paxillin as a component EGF signaling in renal epithelial cells and implicate members of the MAP kinase pathway as critical regulators of paxillin serine/threonine phosphorylation. PMID- 11920685 TI - Activation of apoptosis signal-regulating kinase 1 by the stress-induced activating phosphorylation of pre-formed oligomer. AB - Apoptosis signal-regulating kinase 1 (ASK1) is a MAPKKK family member which activates c-Jun N-terminal kinase (JNK) and p38. In non-stressed cells, ASK1 exists as an inactive complex with the reduced form of thioredoxin. Oxidative stress such as hydrogen peroxide (H2O2) disrupts the ASK1-thioredoxin complex by oxidization of thioredoxin and thereby activates ASK1. The precise mechanism by which ASK1 is activated after its release from thioredoxin is unknown. Here we show that phosphorylation of Thr845 at the activation loop is essential for ASK1 to be activated by H2O2. ASK1 appears to form a silent homo-oligomer through its C-terminal coiled-coil region in non-stressed cells. Following H2O2 treatment, pre-existing ASK1 oligomer undergoes conformational change and creates a new interface within an oligomer, which ultimately leads to trans-autophosphorylation of Thr845. Thus, direct interaction via the coiled-coil region is required for self-scaffolding but not sufficient for activation of ASK1. Importantly, Thr845 of ASK1 can also be trans-phosphorylated by an unidentified Thr845 kinase in response to H2O2 treatment. We propose that this potential Thr845 kinase may be an ignition kinase that triggers Thr845 phosphorylation in oligomerized and activation-competent forms of ASK1. PMID- 11920686 TI - Effects of TGF-alpha gene knockout on epithelial cell kinetics and repair of methotrexate-induced damage in mouse small intestine. AB - While previous studies have indicated that exogenous TGF-alpha stimulates epithelial growth, maintenance, and repair of the gut, roles of endogenous TGF alpha are less well-defined particularly in the small bowel. The current study examined effects of TGF-alpha knockout on adult small intestinal epithelial cell proliferation, migration, apoptosis, and damage/repair response after methotrexate treatment. Compared to normal mice, TGF-alpha gene knockout did not affect crypt cell production, mitosis position, migration, and apoptosis in non injured intestine. RT-PCR gene expression analysis revealed presence of four out of six TGF-alpha related EGF family ligands in the normal intestine, suggesting a possible functional redundancy of the EGF family in maintenance of the intestine. Although TGF-alpha gene knockout did not significantly impair the overall mucosal repair in methotrexate-induced acute damage in the small intestine, it resulted in a higher apoptotic response in the early hours following methotrexate challenge, and a delayed and reduced crypt cell proliferation during repair. Consistently, after methotrexate challenge, intestinal TGF-alpha mRNA was found to be markedly upregulated in the early hours and during repair in the wild type, and there were similar profiles in the increased expression of all other ligands (except EGF) between the wild type and knockout intestines. Therefore, despite a possible functional redundancy among the EGF family ligands in the normal small intestine, TGF-alpha may play a role in modulating the early apoptotic events and in enhancing the subsequent reparative proliferative response in the methotrexate damaged intestine. PMID- 11920687 TI - Nitric oxide is proangiogenic in the retina and choroid. AB - Nitric oxide (NO) has been shown to have proangiogenic or antiangiogenic effects depending upon the setting. In this study, we used mice with targeted deletion of one of the three isoforms of nitric oxide synthase (NOS) to investigate the effects of NO in ocular neovascularization. In transgenic mice with increased expression of vascular endothelial growth factor (VEGF) in photoreceptors, deficiency of any of the three isoforms caused a significant decrease in subretinal neovascularization, but no alteration of VEGF expression. In mice with laser-induced rupture of Bruch's membrane, deficiency of inducible NOS (iNOS) or neuronal NOS (nNOS), but not endothelial NOS (eNOS), caused a significant decrease in choroidal neovascularization. In mice with oxygen-induced ischemic retinopathy, deficiency of eNOS, but not iNOS or nNOS caused a significant decrease in retinal neovascularization and decreased expression of VEGF. These data suggest that NO contributes to both retinal and choroidal neovascularization and that different isoforms of NOS are involved in different settings and different disease processes. A broad spectrum NOS inhibitor may have therapeutic potential for treatment of both retinal and choroidal neovascularization. PMID- 11920688 TI - Factors associated with depression in navy recruits. AB - The age of onset for depression is decreasing and seems to be linked with major life events. This study examined predisposing and concomitant factors related to Navy recruit basic training, and consisted of 443 recruits (200 with depressive symptoms and 243 matched comparison subjects). Two hypotheses were proposed, and both were supported by results of the study. Depressed recruits were more likely to be separated from the Navy and not complete training. They had significantly more predisposing factors than comparison recruits including family history of mental illness, family history of alcohol abuse, and history of psychiatric problems. They also had significantly more concomitant factors such as higher levels of stress, more loneliness, more life-change events, lower sense of belonging, more emotion-oriented coping, and less task-oriented coping. Although the study examined Navy recruits, the findings are applicable to depression in young people. This study adds to understanding the role of life stressors, interpersonal functioning, and development of depressive symptoms. PMID- 11920689 TI - Development of the reasons for living inventory for young adults. AB - Assessment of the reliability, validity, and predictive power of a new measure, the Reasons for Living Inventory for Young Adults (RFL-YA) is described. A series of three studies was conducted at two Midwestern universities to develop initial items for this new measure, refine item selection, and demonstrate the psychometric properties of the RFL-YA. The theoretical differences between the RFL-YA and the College Student Reasons for Living Inventory (CS-RFL) are discussed. Although the two measures were not directly compared, it appears that the RFL-YA has greater specificity for exploring aspects of the protective construct and may be more parsimonious than the CS-RFL. Principal-axis factor analysis yielded a five-factor solution for the RFL-YA accounting for 61.5% of the variance. This five-factor oblique model was confirmed in the final phase of investigation. Alpha estimates for the five subscales ranged from.89 to.94. Concurrent, convergent-discriminant, and criterion validity also were demonstrated. The importance of assessing protective factors in addition to negative risk factors for suicidality is discussed. Directions for future research with the RFL-YA also are discussed. PMID- 11920690 TI - Disclosure to therapists: what is and is not discussed in psychotherapy. AB - This study used the 80-item Disclosure to Therapist Inventory-R to investigate the nature of patient disclosure within therapy. Participants (45 men, 102 women) were all currently in therapy. A Principal Components Analyses with varimax rotation yielded nine meaningful factors; mean disclosure scores were lowest for the factors of Sexuality and Procreation and highest for the factors of Negative Affect and Intimacy. Specific items most extensively discussed included characteristics of parents that are disliked, and aspects of one's personality that are disliked or worrisome. No significant differences were found in overall degree of disclosure as a function of patient gender or shame-proneness; disclosure was, however, found to be positively correlated with strength of the therapeutic alliance. PMID- 11920691 TI - MMPI-2 profiles of Gulf and Vietnam combat veterans with chronic posttraumatic stress disorder. AB - The current study examined service era differences in a sample of 172 Gulf and Vietnam outpatient veterans with combat-related posttraumatic stress disorder (PTSD). Participants completed the MMPI-2 and several additional self-report measures of symptom severity (PTSD, depression, anxiety, hostility, and health complaints). Results indicated that MMPI-2 profiles differed significantly according to service era with Vietnam veterans scoring higher on scales 2, 8, and 0 and lower on scale 9 than did Gulf veterans. Examination of group means derived from parametric analysis of MMPI-2 data suggested a mean two-point code type of 2 8/8-2 for Vietnam veterans and 1-8/8-1 for Gulf veterans. In contrast, when the data were examined using descriptive techniques based on frequency counts of individual MMPI-2 profiles, the most frequently occurring two-point codetype was 7-8/8-7 for Vietnam veterans, and 6-8/8-6 for Gulf veterans. In addition, Gulf veterans reported a greater number of total health complaints than Vietnam veterans, whereas Vietnam veterans reported a greater number of physician diagnosed physical conditions. Potential advantages of incorporating descriptive approaches versus parametric methods when examining profile data are also presented. PMID- 11920692 TI - Psychometric analysis of the cultural mistrust inventory with a Black psychiatric inpatient sample. AB - An important empirical question is whether the Cultural Mistrust Inventory (CMI) can be used to measure cultural aspects of paranoia in Black psychiatric populations. Research on cultural mistrust in psychiatric populations is essential for tests of hypotheses regarding the misdiagnosis of African Americans. The purpose of this study was to assess the reliability and validity of the CMI in a sample of African American patients recently admitted to a psychiatric hospital. The reliability assessment revealed that the CMI total scale is a reliable measure. Factor analytic results indicated that a single global dimension underlies CMI scores. The CMI also showed good convergent validity with a measure of nonclinical paranoia and discriminant validity with measures of self-esteem and social desirability. This study provides reasonable evidence that use of the CMI with psychiatric patients is psychometrically sound. PMID- 11920693 TI - Auld lang syne: success predictors, change processes, and self-reported outcomes of New Year's resolvers and nonresolvers. AB - New Year's resolvers (n = 159) and comparable nonresolvers interested in changing a problem later (n = 123) were followed for six months via telephone interviews to determine their self-reported outcomes, predictors of success, and change processes. The two groups did not differ in terms of demographic characteristics, problem histories, or behavioral goals (weight loss, exercise program, and smoking cessation being the most prevalent). Resolvers reported higher rates of success than nonresolvers; at six months, 46% of the resolvers were continuously successful compared to 4% of the nonresolvers. Self-efficacy, skills to change, and readiness to change assessed before January 1 all predicted positive outcome for resolvers. Once into the new year, successful resolvers employed more cognitive-behavioral processes but fewer awareness-generating and emotion enhancing processes than nonsuccessful resolvers. Discussion centers on the research and intervention opportunities afforded by the annual tradition of resolutions. PMID- 11920694 TI - The Hispanic MMPI-A across five countries. AB - The Hispanic MMPI-A was developed for use with Spanish-speaking adolescents living in the United States. In order to sample its potential usefulness in other countries, the test was administered to 385 Spanish-speaking adolescents in Colombia, Mexico, Peru, Spain, and the United States. The results showed a high degree of similarity across the five countries on the basic content and supplementary scales. Most scales were within 1/2 standard deviation of the U.S. Hispanic mean, and no scale elevations were greater than T = 65. Slight differences (T = 55-60) were obtained on scale F for males and scales F, Hs, Mf (Peru and Colombia), Pa, and Sc for females. This study would suggest that the Hispanic MMPI-A, with its established norms, is appropriate for adaptation in Spanish-speaking countries other than the United States. Further studies with the Hispanic MMPI-A are recommended to determine its effectiveness in clinical assessment in Spanish-speaking countries. PMID- 11920695 TI - Forgiveness and romantic relationships in college: can it heal the wounded heart? AB - This study evaluated the effects of two versions of a six-week group forgiveness intervention for college women who had been wronged in a romantic relationship. Participants (N = 58) were randomly assigned to a secular, religiously integrated, or no-intervention comparison condition. Participants completed a variety of forgiveness and mental health measures at one-week pretest, one-week posttest, and six-week follow-up. Participants in both intervention conditions improved significantly more than did those in the comparison condition on two measures of forgiveness and a measure of existential well-being. Program effects were maintained at six-week follow-up. Contrary to hypotheses, there were no differential treatment effects when comparing participants in the secular and the religiously integrated conditions. Participants generally rated the programs favorably. Forgiveness strategies utilized by participants also were examined. PMID- 11920696 TI - Factor structure, reliability, and validity of the Beck Anxiety Inventory in adolescent psychiatric inpatients. AB - The purpose of this study was to examine further the factor structure, reliability, and validity of the Beck Anxiety Inventory (BAI) in samples of adolescents, ages 14 to 18 years. The BAI is a 21-item self-report measure of anxiety severity. The BAI total score differentiated between the inpatient and high-school adolescents. In the psychiatric inpatient sample, girls obtained higher anxiety-severity scores than boys; no gender differences were obtained for the BAI total score in the high-school sample. Confirmatory factor analyses did not provide adequate fit for the two- factor oblique BAI models to the separate male and female psychiatric inpatient data. Principal axes with varimax and promax rotations initially identified a four-factor solution in the separate male and female inpatient participants. However, second-order analyses of the primary factors provided stronger support for a single-factor structure in each sample. Estimates of reliability for the BAI were adequate in samples of psychiatric inpatient and high-school adolescents. Evidence for convergent and discriminant validity of the BAI was investigated separately in the male and female inpatient samples. Overall, the BAI showed acceptable psychometric properties in these populations. PMID- 11920697 TI - Personality traits and pathology in older and younger incarcerated women. AB - Personality disorders were examined in 157 incarcerated women, using the Schedule for Nonadaptive and Adaptive Personality (SNAP; Clark, 1996) to assess 10 Diagnostic and Statistical Manual of Mental Disorders, Third Edition, Revised (DSM-III-R; American Psychiatric Association, 1987) diagnostic categories and dimensions and 15 domains of trait/temperament relevant to disordered personality. Similar to both community samples and incarcerated men, older women had lower rates of personality disorders than younger women, and the difference was mostly accounted for by differences in the Cluster B disorders. In an exploratory analysis of trait and temperament scales, the older women also scored lower in Aggression, Disinhibition, Entitlement, Exhibitionism, Impulsivity, and Manipulativeness while younger women scored lower in Workaholism and Propriety. These findings suggest that remission of antisocial behavior in women may be associated with changes in lower order personality traits or temperament. PMID- 11920698 TI - MMPI-2 performance of Mexican male university students and prison inmates. AB - In this study, we compared the Minnesota Multiphasic Personality Inventory-2 (MMPI-2) performance of male university students and incarcerated criminal offenders in Mexico. Our aim was to determine whether the MMPI-2 can effectively differentiate between these two distinct groups on scales that are reflective of antisocial behaviors. Our expectations were highly confirmed across the three sets of scales that we considered: the validity and clinical, content, and supplementary scales. Criminal offenders obtained higher or more pathological scores on such scales as Infrequency (F), Schizophrenia (Sc), MacAndrew Alcoholism Scale (MAC-R), Fears (FRS), and so on. Overall, these findings support further research with such target populations as prison inmates in Mexico and other countries in Latin America. PMID- 11920699 TI - The development of a scoring system for the Gerontological Apperception Test. AB - To explore the viability of a revised and more differentiated scoring system for use with the Gerontological Apperception Test (GAT; Wolk & Wolk, 1971), 102 older adults (mean age = 68.3 years) were administered the GAT. Scoring criteria were developed to reflect a variety of constructs speaking to the interpersonal, health-related, and intrapsychic dimensions of the experience of later life. For each of these 20 GAT variables, indices of interscorer agreement exceeded 80%. Bivariate correlations suggested that 12 of 20 GAT dimension scores were related to chronological age as well as to scores derived from measures of crystallized and fluid ability. This also was true for 2 of 5 GAT factor scores. Not only do these data suggest that newly developed GAT variables can be reliably scored, but they also indicate that these more carefully differentiated variables relate to measures of intellectual functioning in theoretically meaningful ways. These findings therefore speak to the utility of the GAT using more carefully defined scoring criteria. PMID- 11920700 TI - The lay concept of "mental disorder" among American undergraduates. AB - Lay concepts of "mental disorder" were investigated in a pilot study of beliefs about 68 conditions, 47 of which corresponded to DSM-IV mental disorders. Undergraduates who had no formal education in abnormal psychology rated the conditions on features proposed in technical definitions of "mental disorder" and judged whether the conditions were mental disorders. The features composed three dimensions-social deviancy, harmful dysfunction, and peculiarity-the last two of which were strongly and independently associated with judgments of mental disorder (R = 0.83). Lay and DSM-IV understandings of "mental disorder" showed moderate convergence. PMID- 11920701 TI - Mast cells in Wallerian degeneration: morphologic and ultrastructural changes. AB - The morphologic and ultrastructural changes of mast cells were followed in degenerating distal and regenerating proximal stumps of frog brachial nerve during Wallerian degeneration. Quantitative analysis included determination of both number and size of mast cells. The mast cell response to injury consisted of an early and a late phase. In the early phase, there was an increase in mast cell numbers in the proximal site of the lesion and a release of Alcian blue material consistent with mediator release. This phase of mast cell activation may be related, through the secretion of biogenic agents such as heparin and histamine, to the increase of endoneurial vessel size and vascular permeability, providing access for macrophages and mast cell precursors. The later phase, which peaked at 40 days after transection in the degenerating distal stump, consisted in the degranulation of the mast cells. These mast cells, closely associated with macrophages and degenerating Schwann cells, released secretory granules into the endoneurial microenvironment. These degranulating mast cells, through the released acid hydrolases, may contribute along with macrophages and Schwann cells, to the degradation of myelin debris. At the same time, mast cells appeared filled with granular content in the regenerating proximal segment. Therefore, we suggest that mast cells in peripheral nerves may play an important role in nerve degenerating and regenerating mechanisms through the secretion of diffusible molecules. PMID- 11920702 TI - Synaptic organization of the mushroom body calyx in Drosophila melanogaster. AB - The calyx neuropil of the mushroom body in adult Drosophila melanogaster contains three major neuronal elements: extrinsic projection neurons, presumed cholinergic, immunoreactive to choline acetyltransferase (ChAT-ir) and vesicular acetylcholine transporter (VAChT-ir) antisera; presumed gamma-aminobutyric acid (GABA)ergic extrinsic neurons with GABA-like immunoreactivity; and local intrinsic Kenyon cells. The projection neurons connecting the calyx with the antennal lobe via the antennocerebral tract are the only source of cholinergic elements in the calyces. Their terminals establish an array of large boutons 2-7 microm in diameter throughout all calycal subdivisions. The GABA-ir extrinsic neurons, different in origin, form a network of fine fibers and boutons codistributed in all calycal regions with the cholinergic terminals and with tiny profiles, mainly Kenyon cell dendrites. We have investigated the synaptic circuits of these three neuron types using preembedding immuno-electron microscopy. All ChAT/VAChT-ir boutons form divergent synapses upon multitudinous surrounding Kenyon cell dendrites. GABA-ir elements also regularly contribute divergent synaptic input onto these dendrites, as well as occasional inputs to boutons of projection neurons. The same synaptic microcircuits involving these three neuron types are repeatedly established in glomeruli in all calycal regions. Each glomerulus comprises a large cholinergic bouton at its core, encircled by tiny vesicle-free Kenyon cell dendrites as well as by a number of GABAergic terminals. A single dendritic profile may thereby receive synaptic input from both cholinergic and GABAergic elements in close vicinity at presynaptic sites with T-bars typical of fly synapses. ChAT-ir boutons regularly have large extensions of the active zones. Thus, Kenyon cells may receive major excitatory input from cholinergic boutons and considerable postsynaptic inhibition from GABAergic terminals, as well as, more rarely, presynaptic inhibitory signaling. The calycal glomeruli of Drosophila are compared with the cerebellar glomeruli of vertebrates. The cholinergic boutons are the largest identified cholinergic synapses in the Drosophila brain and an eligible prospect for studying the genetic regulation of excitatory presynaptic function. PMID- 11920704 TI - Comparison of the basal ganglia in rats, marmosets, macaques, baboons, and humans: volume and neuronal number for the output, internal relay, and striatal modulating nuclei. AB - This study compares the basal ganglia of rats, marmosets, macaques, baboons, and humans. It uses established protocols to estimate the volume and number of neurons within the output nuclei (internal globus pallidus, IGP; and nondopaminergic substantia nigra, SNND), two internal relay and modulating nuclei (subthalamic nucleus, STh; and external globus pallidus, EGP), and a modulator of the striatum (dopaminergic substantia nigra, SND). Nuclear boundaries were defined by using immunohistochemistry for striatal afferents. Total numbers of Nissl-stained and parvalbumin-immunoreactive neurons were calculated by using the fractionator technique. Comparisons between species were standardized relative to brain mass (rats < marmosets < macaques < baboons < humans). The EGP consistently had more neurons relative to the IGP, STh, and SND, which had similar neuronal numbers within each species. The SNND had proportionally more neurons in rats than in primates (especially humans). The distribution of SND neurons varied substantially between rats and primates (very few ventrally located neurons in rats) with humans containing fewer SND neurons than other primates. The reduction in SND neurons in humans suggests less dopaminergic regulation of the basal ganglia system compared with other species. The consistency in the number of IGP neurons across all species, combined with the reduction in SNND neurons in humans, suggests a greater emphasis on output pathways through the IGP and that there are proportionally more STh and EGP neurons in humans. PMID- 11920705 TI - Electrophysiological profile of avian hippocampal unit activity: a basis for regional subdivisions. AB - Electrophysiological activity was recorded from single neurons (units) in the hippocampal formation (HF) of freely moving homing pigeons in order to provide a taxonomy of unit types found in the avian HF; a taxonomy that could be used to define regional subdivisions and be compared with unit types found in the mammalian hippocampus. Two distinct types of unit were observed in the avian HF. One type was uniformly characterized by relatively rapid firing rates and shorter spike widths, and was found throughout the HF. The other type was more variable in activity profile but, compared with the fast-firing units, was characterized by slower firing rates and longer spike widths. However, despite the variable nature of the slow-firing units, most slow-firing units recorded within a given anatomical region displayed similar firing rates, spike widths, and interspike intervals. In general, ventral HF units displayed activity patterns similar to projection cells found in the mammalian Ammon's horn. Most dorsocaudal units displayed activity patterns similar to presumed granular cells in the mammalian dentate gyrus. By contrast, most dorsorostral units displayed activity patterns similar to a type of unit found in the mammalian subiculum. Although different in some details, the overall activity profile of units found in the avian HF, and their regional distribution, is strikingly similar to unit types found in the mammalian hippocampus, suggesting that unit activity profile is one hippocampal dimension conserved through evolution. PMID- 11920706 TI - Role of retinal afferents in regulating growth and shape of the lateral geniculate nucleus. AB - Segregated binocular maps in the dorsal lateral geniculate nucleus (LGN) develop from a stage where they initially overlap. Sophisticated computational models have been used to describe the dynamics of three-dimensional LGN shape changes that play a role in segregation. These models have revealed specific nuclear growth vectors associated with the process of ocular segregation in the LGN (Williams and Jeffery [2001] J Comp Neurol 430:332-342.). In this study, we use similar techniques to determine whether retinal innervation contributes to the dynamics of shape maturation in the ferret LGN. In this animal, 90% of the retinal innervation of the mature LGN comes from the contralateral eye. If one eye is removed before segregation, the projection from the remaining eye remains diffuse and nuclear growth is stunted. Here, we quantify this effect and show that removing the contralateral projection before segregation has a profound impact on LGN size but changes its ultimate shape by only 12%. The impact on shape on the other side of the brain where the ipsilateral projection is removed, which accounts for only 10% of its innervation in maturity, is less than 2%. Hence, retinal innervation plays a minor role in determining mature LGN shape. Although in both hemispheres, the ultimate shape of the nucleus is close to normal, removal of the larger projection disrupts normal growth vectors, with almost none being present in the 5 days after enucleation, when the normal nucleus expands markedly. Hence, the effect of enucleation is to delay shape maturation. Growth vectors absent after removal of the smaller projection are mainly confined to those in what would be the binocular region. PMID- 11920703 TI - Vesicular gamma-aminobutyric acid transporter expression in amacrine and horizontal cells. AB - The vesicular gamma-aminobutyric acid (GABA) transporter (VGAT), which transports the inhibitory amino acid transmitters GABA and glycine, is localized to synaptic vesicles in axon terminals. The localization of VGAT immunoreactivity to mouse and rat retina was evaluated with light and electron microscopy by using well characterized VGAT antibodies. Specific VGAT immunoreactivity was localized to numerous varicose processes in all laminae of the inner plexiform layer (IPL) and to the outer plexiform layer (OPL). Amacrine cell somata characterized by weak VGAT immunoreactivity in the cytoplasm were located in the ganglion cell layer and proximal inner nuclear layer (INL) adjacent to the IPL. In rat retina, VGAT immunoreactive cell bodies also contained GABA, glycine, or parvalbumin (PV) immunoreactivity, suggesting vesicular uptake of GABA or glycine by these cells. A few varicose VGAT-immunoreactive processes entered the OPL from the IPL. VGAT immunoreactivity in the OPL was predominantly localized to horizontal cell processes. VGAT and calcium binding protein-28K immunoreactivities (CaBP; a marker for horizontal cells) were colocalized in processes and terminals distributed to the OPL. Furthermore, VGAT immunoreactivity overlapped or was immediately adjacent to postsynaptic density-95 (PSD-95) immunoreactivity, which is prominent in photoreceptor terminals. Preembedding immunoelectron microscopy of mouse and rat retinae showed that VGAT immunoreactivity was localized to horizontal cell processes and their terminals. Immunoreactivity was distributed throughout the cytoplasm of the horizontal cell processes. Taken together, these findings demonstrate VGAT immunoreactivity in both amacrine and horizontal cell processes, suggesting these cells contain vesicles that accumulate GABA and glycine, possibly for vesicular release. PMID- 11920707 TI - Abnormal positioning of granule cells alters afferent fiber distribution in the mouse fascia dentata: morphologic evidence from reeler, apolipoprotein E receptor 2-, and very low density lipoprotein receptor knockout mice. AB - The fascia dentata of the hippocampal formation is characterized by the nonoverlapping and lamina-specific termination of afferent fibers: entorhinal fibers terminate in the outer molecular layer and commissural/associational fibers terminate in the inner molecular layer. It has been proposed that this fiber lamination depends on the presence of the correct postsynaptic partner at the time of fiber ingrowth during development. Pioneer neurons that guide afferent fibers to their correct layers as well as signals located on granule cells have both been implicated. To study the role of granule cells for the lamina-specific ingrowth of afferents, the cyto- and fiberarchitecture of three mouse mutants (very low density lipoprotein receptor knockout mouse, apolipoprotein E receptor 2 knockout mouse, and reeler mouse) that show different degrees of granule cell migration defects were analyzed. Anterograde tracing with Phaseolus vulgaris-leucoagglutinin was used to visualize the afferent fiber systems, and immunohistochemistry was used to determine the position of their putative target cells. In controls, granule cells are packed in a single layer. This laminar organization is mildly altered in very low density lipoprotein receptor knockout mice, moderately disturbed in apolipoprotein E receptor 2 knockout mice, and severely disrupted in reeler mice. These changes in granule cell distribution are mirrored by the distribution of commissural fibers. In contrast, changes in granule cell distribution do not severely affect the laminar termination of entorhinal fibers. These data provide further evidence for a role of granule cells in the laminar termination of commissural/associational afferents to the fascia dentata. PMID- 11920708 TI - Hypophysiotropic neurons of the paraventricular nucleus respond in spatially, temporally, and phenotypically differentiated manners to acute vs. repeated restraint stress: rapid publication. AB - Hypothalamic-pituitary-adrenal (HPA) responses to stress are initiated by parvicellular neurosecretory neurons in the medial parvicellular (mp) part of the paraventricular hypothalamic nucleus (PVH), which express corticotropin-releasing factor (CRF), among other neuropeptides. We have used an approach guided by patterns of stress-induced Fos expression to explore the manner in which anatomically and phenotypically defined components of the mpPVH respond to acute vs. repeated restraint stress. Hormonal indices of HPA activation in animals exposed to the last of 14 daily repeated restraint sessions were significantly lower than those in rats receiving a single restraint episode. Although this habituation was paralleled by global decrements in activation patterns across all PVH compartments, clear spatial-temporal differences in recruitment profiles were noted between dorsal and ventral aspects of the mpPVH. Thus, acute restraint provoked a biphasic Fos induction, which occurred first within the mpPVH and in an adjoining population of somatostatinergic cells in the periventricular region and only later within other aspects of the PVH. By contrast, Fos responses of habituated animals were monophasic and focused decisively within a discrete ventral aspect of the mpPVH. The ventral population was identified as comprising neurons that express CRF and/or enkephalin and, to a lesser extent, growth hormone-releasing factor. These results indicate a lack of homogeneity among stress-responsive parvicellular neurosecretory neurons and suggest that distinct complements of CRF cells may be preferentially involved in initiating HPA responses to acute stress and sustaining them in the repeated condition. PMID- 11920709 TI - Temporal progression of angiogenesis and basal lamina deposition after contusive spinal cord injury in the adult rat. AB - After spinal cord injury (SCI), the absence of an adequate blood supply to injured tissues has been hypothesized to contribute to the lack of regeneration. In this study, blood vessel changes were examined in 28 adult female Fischer 344 rats at 1, 3, 7, 14, 28, and 60 days after a 12.5 g x cm NYU impactor injury at the T9 vertebral level. Laminin, collagen IV, endothelial barrier antigen (SMI71), and rat endothelial cell antigen (RECA-1) immunoreactivities were used to quantify blood vessel per area densities and diameters in ventral gray matter (VGM), ventral white matter (VWM), and dorsal columns (DC) at levels ranging 15 mm rostral and caudal to the epicenter. This study demonstrates an angiogenic response, defined as SMI71/RECA-1-immunopositive endothelial cells that colocalize with a robust deposition of basal lamina and basal lamina streamers, 7 days after injury within epicenter VGM. This angiogenesis diminishes concurrent with cystic cavity formation. GAP43- and neurofilament- (68 kDa and 210 kDa) immunopositive fiber outgrowth was associated with these new blood vessels by day 14. Between 28 and 60 days after injury, increases in SMI71-immunopositive blood vessel densities were observed in the remaining VWM and DC with a corresponding increase in vessel diameters up to 15 mm rostral and caudal to the epicenter. This second angiogenesis within VWM and DC, unlike the acute response observed in VGM, did not correspond to any previously described changes in locomotor behaviors in this model. We propose that therapies targeting angiogenic processes be directed at the interval between 3 and 7 days after SCI. PMID- 11920710 TI - Light- and electron-microscopic analysis of vasoactive intestinal polypeptide immunoreactive amacrine cells in the guinea pig retina. AB - Vasoactive intestinal polypeptide (VIP) is a neuroactive substance that is expressed in both nonmammalian and mammalian retinas. This study investigated the morphology and synaptic connections of VIP-containing neurons in the guinea pig retina by immunocytochemistry, by using antisera against VIP. Specific VIP immunoreactivity was localized to a population of wide-field and regularly spaced amacrine cells with processes ramifying mainly in strata 1 and 2 of the inner plexiform layer (IPL). Double-label immunohistochemistry demonstrated that all VIP-immunoreactive cells possessed gamma-aminobutyric acid immunoreactivity. The synaptic connectivity of VIP-immunoreactive amacrine cells was identified in the IPL by electron microscopy. The VIP-labeled amacrine cell processes received synaptic input from other amacrine cell processes and bipolar cell axon terminals in strata 1 to 3 of the IPL. The most frequent postsynaptic targets of VIP immunoreactive amacrine cells were other amacrine cell processes in strata 1 to 3 of the IPL. Synaptic outputs to bipolar cells were also observed in strata 1 to 3 of the IPL. In addition, ganglion cell dendrites were also postsynaptic to VIP immunoreactive neurons in the sublamina a of the IPL. These studies show that one type of VIP-immunoreactive amacrine cells make contact predominantly with other amacrine cell processes. This finding suggests that VIP-containing amacrine cells may influence inner retinal circuitry, thus mediating visual processing. PMID- 11920711 TI - Projections of the sexually dimorphic calcitonin gene-related peptide neurons of the preoptic area determined by retrograde tracing in the female rat. AB - The medial preoptic area of the rat exhibits morphologic sex differences and is implicated in the control of sexually dimorphic behavior and function. Neurons expressing calcitonin gene-related peptide (CGRP) within the anteroventral periventricular (AVPV) and medial preoptic nucleus (MPN) of the medial preoptic area exhibit female-dominant sex differences in number through organizational and activational effects of gonadal steroids. The present study used retrograde tracing experiments to establish the projections of the AVPV and MPN CGRP neurons in the female rat. After the intraperitoneal administration of Fluoro-Gold to female rats (n = 5), we were unable to detect retrograde tracer in any CGRP immunoreactive cells of the hypothalamus. Intracerebral injections of 50- to 100 nl volumes of Fluoro-Gold into the mediobasal hypothalamus resulted in up to 70% of CGRP neurons in the AVPV and MPN containing retrograde tracer. Similar large volume tracer depositions in the lateral septum, periaqueductal gray, two likely CGRP projection sites, resulted in no labeling of preoptic CGRP neurons. Experiments using small volume (30-nl) injections of Fluoro-Gold and green fluorescent microspheres at multiple sites in the mediobasal hypothalamus (n = 18) revealed that approximately 60% of AVPV and 30% of MPN neurons expressing CGRP were projecting to the region of the tuberal and ventral premammillary nuclei, with a minor projection to the dorsomedial nucleus. These findings demonstrate a major projection of the preoptic CGRP neurons to the posterior hypothalamus in the female rat and support further a functional role for these neurons in the sexually dimorphic regulation of reproductive functioning. PMID- 11920712 TI - Distribution and terminal arborizations of cutaneous mechanoreceptors in the glabrous finger pads of the monkey. AB - Recent electrophysiological studies demonstrated that neurons in the somatosensory cortex of monkeys respond to tangential forces applied to glabrous skin. To unravel the peripheral basis for this cortical response, we determined the distribution of presumptive low-threshold mechanoreceptors innervating the distal finger pads of monkeys. Endings were reconstructed in immunolabeled serial sections imaged by epifluorescence and confocal microscopy. Although classically implicated as cutaneous stretch receptors, no Ruffini corpuscles were found in the glabrous skin. Ruffini-like endings were only detected at the base of the finger nails. Pacinian corpuscles were sparsely distributed in the deep dermis. Meissner corpuscles (MCs) in dermal papillary ridges had a comparably high density in the thumb, index, and fifth fingers. Each MC was innervated by several large-caliber axons. Within the limits of our reconstructions, some of these axons terminated in only one MC, whereas others innervated several MCs. Merkel endings covered about 80% of the base of the intermediate epidermal ridges that form the pattern of fingerprints. In some cases, the distal tip of a Merkel related axon gave rise to a several terminal branches that supplied endings to tightly circumscribed (30-70 microm) clusters of Merkel cells. In other cases, the nodes of axons gave rise to en passant branches that formed extended chains of endings among Merkel cells spread over territories up to 300 microm long. Based on their relatively diffuse distributions, the axons that innervate multiple MCs or the axons with en passant Merkel terminations seem most suited to transduce tangential forces. PMID- 11920713 TI - Axon arbors and synaptic connections of a vulnerable population of interneurons in the dentate gyrus in vivo. AB - The predominant gamma-aminobutyric acid (GABA)ergic neuron class in the hilus of the dentate gyrus consists of spiny somatostatinergic interneurons. We examined the axon projections and synaptic connections made by spiny hilar interneurons labeled with biocytin in gerbils in vivo. Axon length was 152-497 mm/neuron. Sixty to 85% of the axon concentrated in the outer two thirds of the molecular layer of the dentate gyrus. The septotemporal span of the axon arbor extended over 48-82% of the total hippocampal length, which far exceeds the septotemporal span of axons of granule cells whose complete axon arbors extended over 15-29%. A three-dimensionally reconstructed 216-microm-long spiny hilar interneuron axon segment in the outer third of the molecular layer formed an average of 1 synapse every 5.1 microm. Of the 42 symmetric (inhibitory) synapses formed by the reconstructed segment, 88% were with spiny dendrites of presumed granule cells, and 67% were with dendritic spines that also receive an asymmetric (excitatory) contact from an unlabeled axon terminal. Postembedding GABA-immunocytochemistry revealed that 55% of the GABAergic synapses in the outer third of the molecular layer were with spines. Therefore, in the outer molecular layer, spiny hilar interneurons form synaptic contacts that appear to be positioned to exert inhibitory control near sites of excitatory synaptic input from the entorhinal cortex to granule cell dendritic spines. These findings demonstrate far-reaching, yet highly specific, connectivity of individual interneurons and suggest that the loss of spiny hilar interneurons, as occurs in temporal lobe epilepsy, may contribute to hyperexcitability in the hippocampus. PMID- 11920714 TI - Adult-like complexity of the larval antennal lobe of D. melanogaster despite markedly low numbers of odorant receptor neurons. AB - We provide a detailed analysis of the larval head chemosensory system of Drosophila melanogaster, based on confocal microscopy of cell-specific reporter gene expression in P[GAL4] enhancer trap lines. In particular, we describe the neuronal composition of three external and three pharyngeal chemosensory organs, the nerve tracts chosen by their afferents, and their central target regions. With a total of 21 olfactory and 80 gustatory neurons, the sensory level is numerically much simpler than that of the adult. Moreover, its design is different than in the adult, showing an association between smell and taste sensilla. In contrast, the first-order relay of the olfactory afferents, the larval antennal lobe (LAL), exhibits adult-like features both in terms of structure and cell number. It shows a division into approximately 30 subunits, reminiscent of glomeruli in the adult antennal lobe. Taken together, the design of the larval chemosensory system is a "hybrid," with larval-specific features in the periphery and central characteristics in common with the adult. The largely reduced numbers of afferents and the similar architecture of the LAL and the adult antennal lobe, render the larval chemosensory system of Drosophila a valuable model system, both for studying smell and taste and for examining the development of its adult organization. PMID- 11920715 TI - Retinal organization in the bcl-2-overexpressing transgenic mouse. AB - Naturally occurring cell death is believed to play a major role during the development of the nervous system in the establishment of neuronal architecture. Here we study the effects of cell death inhibition by using a transgenic mouse in which the powerful antiapoptotic gene bcl-2 is expressed in neurons. The retina of this mouse reveals that the general neuronal plan has been maintained. However, bcl-2 overexpression leads to altered frequencies of the major cell types in the retina. Thus, it is possible to estimate cell-type-specific rates of apoptosis by observing the increases in numbers of cells in the bcl-2 overexpressing transgenic mouse. PMID- 11920716 TI - Descending supraspinal pathways in amphibians: III. Development of descending projections to the spinal cord in Xenopus laevis with emphasis on the catecholaminergic inputs. AB - In developmental stages of the clawed toad, Xenopus laevis, we describe the ontogeny of descending supraspinal connections, catecholaminergic projections in particular, by means of retrograde tracing techniques with dextran amines. Already at embryonic stages (stage 40), spinal projections from the reticular formation, raphe nuclei, Mauthner neurons, vestibular nuclei, the locus coeruleus, the interstitial nucleus of the medial longitudinal fasciculus, the posterior tubercle, and the periventricular nucleus of the zona incerta are well developed. At the beginning of the premetamorphic period (stage 46), spinal projections arise from the suprachiasmatic nucleus, the torus semicircularis, the pretectal region, and the ventral telencephalon. After stage 48, tectospinal and cerebellospinal projections develop, with spinal projections from the preoptic area following at stage 51. Rubrospinal projections are present at stage 50. During the prometamorphic period, spinal projections arise in the nucleus of the solitary tract, the lateral line nucleus, and the mesencephalic trigeminal nucleus. With in vitro double-labeling methods, based on retrograde tracing of dextran amines in combination with tyrosine hydroxylase (TH) immunohistochemistry, we show that at stage 40/41, catecholaminergic (CA) neurons in the posterior tubercle are the first to project to the spinal cord. Subsequently, at stage 43, new projections arise in the periventricular nucleus of the zona incerta and the locus coeruleus. The last CA projection to the spinal cord originates from neurons in the nucleus of the solitary tract at the beginning of prometamorphosis (stage 53). Our data show a temporal, rostrocaudal sequence in the development of the CA cell groups projecting to the spinal cord. Moreover, the early appearance of CA fibers, preterminals and terminal-like structures in dorsal, intermediate, and ventral zones of the embryonic spinal cord, suggests an important role for catecholamines during development in nociception, autonomic functions, and motor control at the spinal level. PMID- 11920717 TI - Central organization of the electrosensory system in the paddlefish (Polyodon spathula). AB - The central connections of the electrosensory system were studied in the paddlefish Polyodon spathula by injecting biotinylated dextran amines into the dorsal octavolateral nucleus (DON), the cerebellum, and the mesencephalic tectum. The sole target of primary electrosensory fibers is the ipsilateral dorsal octavolateral nucleus. The principal neurons ascending from this nucleus project to the torus semicircularis, the lateral mesencephalic nucleus, and the mesencephalic tectum. The mesencephalic tectum projects back to the nucleus preeminentialis, which, in turn, projects to the cerebellar auricles and to the DON. The auricles are the main source of parallel fibers in the cerebellar crest ventral to the DON. The DON also receives input from the contralateral DON. These descending feedback loops are very similar to those of other electrosensory fishes. However, the paddlefish is unique in having three mesencephalic targets of electrosensory information. It is the only bony fish known to have extensive projections directly to the mesencephalic tectum and to a lateral mesencephalic nucleus in addition to the torus semicircularis. PMID- 11920718 TI - Calcyon in the rat brain: cloning of cDNA and expression of mRNA. AB - Calcyon is a 24 kD protein recently cloned from a human brain cDNA library and shown to interact with the dopamine receptor 1 (D1R) of D1-like receptors. This interaction shifts the effector coupling of D1R to stimulate a calcium signaling pathway, without influencing the D1R-adenylyl-cAMP pathway. To obtain more knowledge about the potential role of calcyon in the brain, we cloned rat calcyon cDNA and studied its distribution in the brain. Northern blot analysis and RT-PCR revealed that rat calcyon mRNA was expressed only in the brain. With the use of the in situ hybridization technique, we studied rat calcyon mRNA distribution in the brain and related it to the distribution of D1R and dopamine receptor 5 (D5R) mRNAs. Prominent calcyon mRNA signals were found in several brain regions, including hippocampus, hypothalamus, cerebellum, and medial prefrontal cortex. Less abundant calcyon mRNA expression was observed in the dorsal striatum region, where D1R mRNA is highly expressed and where D1R/cAMP-DARPP-32 signaling pathway is of great functional importance. The strongest expression of D5R mRNA was found in the hippocampus and cerebellum, where D1R mRNA expression was relatively low. In conclusion, rat calcyon appears to be a brain specific protein. There is a certain overlap between calcyon mRNA distribution and that of the D1R and D5 mRNAs, indicating that calcyon might be associated not only with D1R but also with D5R. PMID- 11920719 TI - Distribution and morphology of transgenic mouse oligodendroglial-lineage cells following transplantation into normal and myelin-deficient rat CNS. AB - Glial cells from neonatal MbetaP5 transgenic mice, which express bacterial beta galactosidase (lacZ) under control of the myelin basic protein (MBP) promoter (Gow et al, 1992), were transplanted into the spinal cord or cerebral hemisphere of immunosuppressed normal and myelin-deficient (md) rats in order to assess the ability of the donor cells to survive, migrate, and differentiate within normal compared with myelin-deficient central nervous system (CNS). LacZ+ cells were detected as early as 6-7 days after transplantation into the low thoracic cord and by 10 days had spread rostrally to the brainstem and caudally to the sacral spinal cord. Initially, compact lacZ+ cells, lacking processes, were found associated with small blood vessels and with the glia limitans. Cells of this type persisted throughout the experiment. Later, lacZ+ cells with processes were seen along fiber tracts in the dorsal columns and, after intracerebral injection, subjacent to ventricular ependyma, as well as scattered in cerebral white and gray parenchyma. The extent of spread was comparable in md and normal rats, but in the md group, the success rate was higher, and more cells differentiated into process-bearing oligodendrocytes. Acceptance of xenografts in immunosuppressed recipients equaled that of allografts. The overall spread of grafted cells exceeded that of injected charcoal, indicating active migration. In contrast to earlier studies that identified oligodendrocytes based on morphology alone, this study has allowed us to identify and track oligodendrocytes based on myelin gene expression. We show some oligodendrocytes whose morphology is consistent with classical morphological descriptions, some that resemble astrocytes, and a class of compact perivascular oligodendrocyte-lineage cells that we suggest are migratory. PMID- 11920721 TI - Sublaminar organization of the mouse olfactory bulb nerve layer. AB - Olfactory sensory neuron (OSN) axons coalesce to form the olfactory nerve (ON) and then grow from the olfactory epithelium to the olfactory bulb (OB), enter the olfactory nerve layer (ONL), reorganize extensively, and innervate specific glomeruli. Within the ON and ONL a population of glial cells, the olfactory ensheathing cells (OECs), surround OSN axon fascicles. To better understand the relationship between OECs and axon fascicles in the ONL of the adult mouse, we used confocal microscopy and antibodies to the low affinity nerve growth factor receptor p75 (p75), glial fibrillary acidic protein (GFAP), neuropeptide Y (NPY), and S-100 to identify glia. Antibodies to olfactory marker protein (OMP) and neuronal cell adhesion molecule (NCAM) were used to identify OSN axons. Electron microscopy characterized the ONL ultrastructure. We found that glial processes were not uniformly distributed in the ONL of the mouse. The p75(+) OEC processes were restricted to the ON and the outer ONL sublamina, and oriented parallel to the plane of the OB layers. In the inner ONL NPY(+) OEC-like processes were seen. GFAP(+) processes were restricted to the inner ONL sublamina, the ONL/GL boundary, and the GL, where they delineated loosely aggregated axon fascicles that entered the glomeruli obliquely. S-100(+) processes and somata were distributed throughout the ONL; the outer and inner ONL were equivalent in their S-100 staining. Ultrastructural studies showed that, although OECs could be identified in both the outer and inner ONL, in the latter, their relationship to bundles of OEC axons appeared less orderly than seen in the outer ONL. Our data demonstrate a differential organization of the ONL that could subserve distinct functions; axon extension may occur predominantly in the outermost ONL, whereas glomerular targeting occurs in the inner sublamina of the ONL. PMID- 11920720 TI - Functional aspects of dopamine metabolism in the putative prefrontal cortex analogue and striatum of pigeons (Columba livia). AB - Dopamine (DA) in mammalian associative structures, such as the prefrontal cortex (PFC), plays a prominent role in learning and memory processes, and its homeostasis differs from that of DA in the striatum, a sensorimotor region. The neostriatum caudolaterale (NCL) of birds resembles the mammalian PFC according to connectional, electrophysiological, and behavioral data. In the present study, DA regulation in the associative NCL and the striatal lobus parolfactorius (LPO) of pigeons was compared to uncover possible differences corresponding to those between mammalian PFC and striatum. Extracellular levels of DA and its metabolites (homovanillic acid [HVA], dihydroxyphenylacetic acid [DOPAC]) and the serotonin metabolite 5-hydroxyindoleacetic acid (5-HIAA) were investigated by in vivo microdialysis of urethane-anesthetized pigeons under basal conditions and after systemic administration of D-amphetamine. DA was reliably determined only in LPO dialysates, and DA metabolite levels were significantly higher in LPO than in NCL. The HVA/DOPAC ratio, indicating extracellular lifetime of DA, was more than twice as high in NCL than in LPO dialysates. After amphetamine, DA increased in LPO while still being undetectable in NCL, and DA metabolites decreased in both regions. 5-HIAA slightly decreased in NCL dialysates. Amphetamine effects were delayed in NCL compared with the striatum. In conclusion, effects of amphetamine on the pigeon's ascending monoamine systems resemble those found in mammals, suggesting similar regulatory properties. The neurochemical differences between NCL and LPO parallel those between associative regions, such as PFC and dorsal striatum in mammals. They may reflect weaker regulation of extracellular DA, favoring DAergic volume transmission, in associative than striatal forebrain regions. PMID- 11920722 TI - Axonal projections of pulmonary slowly adapting receptor relay neurons in the rat. AB - We elucidated efferent projections of second-order relay neurons (P-cells) activated by afferents originating from slowly adapting pulmonary receptors (SARs) to determine the central pathway of the SAR-evoked reflexes. Special attention was paid to visualizing the P-cell projections within the nucleus tractus solitarii (NTS), which may correspond to the inhibitory pathway from P cells to second-order relay neurons (RAR-cells) of rapidly adapting pulmonary receptors. P-cells were recorded from the NTS in Nembutal-anesthetized, paralyzed, and artificially ventilated rats. First, we used electrophysiological methods of antidromic mapping and showed that the majority of the P-cells examined projected their axons to the caudal NTS and to the dorsolateral pons corresponding to the parabrachial complex. Second, a mixture of HRP and Neurobiotin was injected intracellularly or juxtramembranously into P-cells. (1) Stained P-cells (n = 7) were located laterally to the solitary tract and had dendrites extending characteristically along the lateral border of the solitary tract. (2) All P-cells had stem axons projecting to the ipsilateral medulla. Of these, the axons from five P-cells projected to the nucleus ambiguus and its vicinity with distributing boutons. Some of these axons further ascended in the ventrolateral medulla, and distributed boutons in the areas ventral or ventrolateral to the nucleus ambiguus. (3) All the P-cells had axonal branches with boutons in the NTS area. In particular, axons from three P-cells projected bilaterally to the medial NTS caudal to the obex, i.e., to the area of RAR-cells. These results show anatomic substrates for the connections implicated in the P cell inhibition of RAR-cells as well as the SAR-induced respiratory reflexes. PMID- 11920723 TI - Antiandrogen blocks estrogen-induced masculinization of the song system in female zebra finches. AB - Song behavior and the neural song system that serves it are sexually dimorphic in zebra finches. In this species, males sing and females normally do not. The sex differences in the song system include sex differences in the proportion of neurons that express androgen receptors, which is higher in specific brain regions of males. Estradiol (E2) administered in early development profoundly masculinizes the song system of females, including the proportion of neurons expressing androgen receptors. We examined whether or not the expression of these androgen receptors was causally related to the E2-induced masculinization of this system by co-administering Flutamide, which blocks androgen action at the receptor, along with E2 at hatching. E2 alone had its usual masculinizing effect on the female song system, measured in adulthood: increasing the size of song nuclei, the size of neurons in HVC, RA, and 1MAN, and the number of neurons in HVC. E2's masculinizing action, however, was significantly diminished on all measures by co-administering Flutamide. Indeed, females receiving both E2 and Flutamide were never significantly more masculine than controls on any measure. Flutamide alone had no effect. Our results strongly suggest that the activation of androgen receptors is necessary for the E2-induced masculinization of the song system in females. PMID- 11920724 TI - Long-term culture of mouse cortical neurons as a model for neuronal development, aging, and death. AB - A long-term cell culture system was used to study maturation, aging, and death of cortical neurons. Mouse cortical neurons were maintained in culture in serum-free medium (Neurobasal supplemented with B27) for 60 days in vitro (DIV). The levels of several proteins were evaluated by immunoblotting to demonstrate that these neurons matured by developing dendrites and synapses and remained continuously healthy for 60 DIV. During their maturation, cortical neurons showed increased or stable protein expression of glycolytic enzyme, synaptophysin, synapsin IIa, alpha and beta synucleins, and glutamate receptors. Synaptogenesis was prominent during the first 15 days and then synaptic markers remained stable through DIV60. Very early during dendritic development at DIV3, beta-synuclein (but not alpha synuclein) was localized at the base of dendritic growth cones identified by MAP2 and alpha-amino-3-hydroxy-5-methyl-4-isoxazole (AMPA) receptor GluR1. In mature neurons, alpha and beta synucleins colocalized in presynaptic axon terminals. Expression of N-methyl-D-aspartate (NMDA) and AMPA receptors preceded the formation of synapses. Glutamate receptors continued to be expressed strongly through DIV60. Cortical neurons aging in vitro displayed a complex profile of protein damage as identified by protein nitration. During cortical neuron aging, some proteins showed increased nitration, while other proteins showed decreased nitration. After exposure to DNA damaging agent, young (DIV5) and old (DIV60) cortical neurons activated apoptosis mechanisms, including caspase-3 cleavage and poly(ADP)-ribose polymerase inactivation. We show that cultured mouse cortical neurons can be maintained for long term. Cortical neurons display compartmental changes in the localization of synucleins during maturation in vitro. These neurons sustain protein nitration during aging and exhibit age-related variations in the biochemistry of neuronal apoptosis. PMID- 11920725 TI - Progesterone receptors and the sexual differentiation of the medial preoptic nucleus. AB - The central component of the medial preoptic nucleus (MPNc) of the rat has served as an excellent model of sexual differentiation. The MPNc is larger in adult males than in females, and its development is regulated by perinatal gonadal hormones. Although testosterone (T) and its metabolite estradiol (E) sexually differentiate this region, the exact mechanism by which they act during development is not known. There is a dramatic sex difference in the expression of progesterone receptors (PR) in the MPN during development; perinatal males express higher levels of PR than females. Additionally, PR expression during this time is dependent on exposure to T. Thus, PR induction may be one mechanism by which T sexually differentiates the MPN. The present study investigated the potential role of PR in the sexual differentiation of the MPNc. Anatomical examination of PR distribution within the MPN of neonatal males revealed the presence of PR immunoreactive cells within the MPNc, suggesting a direct route of action for PR in the development of the MPNc. Additionally, we measured the effects of neonatal RU486 treatment, a progesterone and glucocorticoid receptor antagonist, on subsequent MPNc volume in neonatally T-treated females and neonatally castrated males, given T. RU486 treatment reduced the MPNc volume of T treated females while it increased the volume in T-treated, neonatally castrated males. These results, taken together with the expression of PR in the MPNc, suggest that PR may influence the sexual differentiation of the MPNc volume. PMID- 11920726 TI - Perineal muscles and motoneurons are sexually monomorphic in the naked mole-rat (Heterocephalus glaber). AB - Naked mole-rats are eusocial mammals that live in colonies with a single breeding female and one to three breeding males. All other members of the colony, known as subordinates, are nonreproductive and exhibit few sex differences in behavior or genital anatomy. This raises questions about the degree of sexual differentiation in subordinate naked mole-rats. The striated perineal muscles associated with the phallus [the bulbocavernosus (BC), ischiocavernosus (IC), and levator ani (LA) muscles], and their innervating motoneurons, are sexually dimorphic in all rodents examined to date. We therefore asked whether perineal muscles and motoneurons were also sexually dimorphic in subordinate naked mole-rats. Muscles similar to the LA and IC of other rodents were found in naked mole-rats of both sexes. No clear BC muscle was identified, although a large striated muscle associated with the urethra in male and female naked mole-rats may be homologous to the BC of other rodents. There were no sex differences in the volumes of the LA, IC, or the urethral muscles. Motoneurons innervating the perineal muscles were identified by retrograde labeling with cholera-toxin-conjugated horseradish peroxidase. All perineal motoneurons were found in a single cluster in the ventrolateral lateral horn, in a position similar to that of Onuf's nucleus of carnivores and primates. There was no sex difference in the size or number of motoneurons in Onuf's nucleus of naked mole-rats. Thus, unlike findings in any other mammal, neither the perineal muscles nor the perineal motoneurons appear to be sexually differentiated in subordinate naked mole-rats. PMID- 11920728 TI - Regulated appearance of NMDA receptor subunits and channel functions during in vitro neuronal differentiation. AB - The schedule of NMDA receptor subunit expression and the appearance of functional NMDA-gated ion channels were investigated during the retinoic acid (RA) induced neuronal differentiation of NE-4C, a p53-deficient mouse neuroectodermal progenitor cell line. NR2A, NR2B, and NR2D subunit transcripts were present in both nondifferentiated and neuronally differentiated cultures, while NR2C subunits were expressed only transiently, during the early period of neural differentiation. Several splice variants of NR1 were detected in noninduced progenitors and in RA-induced cells, except the N1 exon containing transcripts that appeared after the fourth day of induction, when neuronal processes were already formed. NR1 and NR2A subunit proteins were detected both in nondifferentiated progenitor cells and in neurons, while the mature form of NR2B subunit protein appeared only at the time of neuronal process elongation. Despite the early presence of NR1 and NR2A subunits, NMDA-evoked responses could be detected in NE-4C neurons only after the sixth day of induction, coinciding in time with the expression of the mature NR2B subunit. The formation of functional NMDA receptors also coincided with the appearance of synapsin I and synaptophysin. The lag period between the production of the subunits and the onset of channel function suggests that subunits capable of channel formation cannot form functional NMDA receptors until a certain stage of neuronal commitment. Thus, the in vitro neurogenesis by NE-4C cells provides a suitable tool to investigate some inherent regulatory processes involved in the initial maturation of NMDA receptor complexes. PMID- 11920727 TI - Temporal regulation of neuropilin-1 expression and sensitivity to semaphorin 3A in NGF- and NT3-responsive chick sensory neurons. AB - The extracellular molecule semaphorin 3A (Sema3A) is proposed to be a negative guidance cue that participates in patterning DRG sensory axons in the developing chick spinal cord. During development Sema3A is first expressed throughout the spinal cord gray matter, but Sema3A expression later disappears from the dorsal horn, where small-caliber cutaneous afferents terminate. Sema3A expression remains in the ventral horn, where large-muscle proprioceptive afferents terminate. It has been proposed that temporal changes in the sensitivity of different classes of sensory afferents to Sema3A contribute to the different pathfinding of these sensory afferents. This study compared the expression of the semaphorin 3A receptor subunit, neuropilin-1, and the collapse response of growth cones to semaphorin 3A for NGF (cutaneous)- and NT3 (proprioceptive)-dependent sensory axons extended from E6-E10 chick embryos. Growth cones extended from E6 DRGs in NT3-containing medium expressed neuropilin-1 and collapsed in response to Sema3A. From E7 until E10 NT3-responsive growth cones expressed progressively lower levels of neuropilin-1, and were less sensitive to Sema3A. On the other hand, growth cones extended from DRGs in NGF-containing medium expressed progressively higher levels of neuropilin-1 and higher levels of collapse response to Sema3A over the period from E6-E10. Thus, developmental patterning of sensory terminals in the chick spinal cord may arise from changes in both Sema3A expression in the developing spinal cord and accompanying changes in neuronal expression of the Sema3A receptor subunit, neuropilin-1. PMID- 11920729 TI - Inositol 1,4,5-trisphosphate transduction cascade in taste reception of the fleshfly, Boettcherisca peregrina. AB - The role of an inositol 1,4,5-trisphosphate (IP3)-mediated transduction cascade in the response of taste receptor cells of the fleshfly Boettcherisca peregrina was investigated by using the following reagents: neomycin (an inhibitor of IP3 production), U73122 (an inhibitor of phospholipase C), adenophostin A (an agonist of the IP3-gated channel), IP3, ruthenium red (a blocker of the IP3-gated channel), and 2-aminoethoxydiphenylborate (2-APB; an antagonist of the IP3-gated channel). For introduction into the receptor cell, the reagents were mixed with a detergent, deoxycholate (DOC). After treatment with neomycin + DOC or U73122 + DOC, the response of the sugar receptor cell to sugars was depressed compared with responses after treatment with DOC alone. During the treatment of adenophostin A + DOC, the response of the sugar receptor cell was elicited. After treatment with IP3 + DOC, the response of the sugar receptor cell to sugars and to amino acids was apparently enhanced. When taste stimuli were administered in the presence of ruthenium red or 2-APB, the response of the sugar receptor cell to glucose were inhibited. The expression of genes for substances involved in the IP3 transduction cascade, such as G protein alpha subunit (dGqalpha), phospholipase C (norpA), and IP3 receptor (itpr), were examined in the taste receptor cell of the fruitfly Drosophila melanogaster by using the pox-neuro70 mutant (poxn70), which lacks taste receptor cells. The expressed levels of dGqalpha and itpr in the tarsus of poxn70 mutant flies were reduced compared with those of wild-type flies. These results suggest that the IP3 transduction cascade is involved in the response of the sugar receptor cell of the fly. PMID- 11920730 TI - Markers of senescence? AB - The specific identification of cellular senescence in clinical material has important implications for determining the role of senescence in age-related pathologies and in neoplasia in certain tumours. One suggested marker of senescence is the histochemical identification of a specific beta-galactosidase enzyme operative at pH 6.0. However, recent data indicate that this enzyme may not be specific for senescence in all tissues and probably represents the expression of endogenous lysosomal acid beta-galactosidase, which is expressed by a variety of differentiated cell types. PMID- 11920731 TI - Current understanding of cellular and molecular events in intervertebral disc degeneration: implications for therapy. AB - Until recently, material removed from the intervertebral disc (IVD) at surgery consisted either of 'loose bodies' from the centre of the IVD or discal tissue displaced (prolapsed) into the intervertebral root or spinal canals. This material is best regarded as a by-product of disc degeneration and therefore not representative of the disease process itself. Recent advances in surgical techniques, particularly anterior fusion, in which large segments of the anterior part of the IVD are excised with the anatomical relationships between different components intact, have generated material that can be investigated with modern molecular and cell biological techniques. This is an important area of study because degeneration of the lumbar IVDs is associated, perhaps causally, with low back pain, one of the most common and debilitating conditions in the West. 'Degeneration' carries implications of inevitable progression of wear-and-tear associated conditions. Modern research on human IVD tissue has shown that this is far from the case and that disruption of the micro-anatomy described as degeneration is an active process, regulated by locally produced molecules. The exciting consequence of this observation is the possibility of being able to inhibit or even reverse the processes of degeneration using targeted therapy. PMID- 11920732 TI - Identification of loci associated with putative recurrence genes in transitional cell carcinoma of the urinary bladder. AB - Following an earlier study linking monosomy 9 with recurrence of transitional cell carcinomas (TCCs) of the urinary bladder, 109 primary and recurrent TCCs (from 47 patients) were examined to explore genetic alterations at chromosome 9 associated with recurrence. Patient DNA was microdissected and extracted from archival tissue sections and analysed for loss of heterozygosity (LOH) at three regions on chromosome 9 where tumour suppressor genes (TSGs) are known to reside (INK 4A, DBC1, and TSC1). Patients were categorized into two groups, non recurrent TCC (NR, n=18) and recurrent TCC (REC, n=29). It was noted that 12% of NR tumours, compared with 54% of REC primary tumours (p=0.01), had LOH at all informative markers spanning the TSC1 region. The risk of recurrence was significantly higher in patients with deleted TSC1 than in those who retained the TSC1 region (p=0.035). Levels of LOH at DBC1 or INK 4A were not significantly different in NR tumours than in REC primary tumours and recurrence-free survival was not affected by loss of either of these genes. Loss of all informative markers spanning chromosome 9 was observed in 0% of NR tumours compared with 25% of REC primary tumours (p=0.04). The probability of recurrence was also significantly increased in patients who had LOH at all informative markers spanning chromosome 9 (p=0.016), confirming earlier fluorescence in situ hybridization results. This study provides further evidence that recurrence in bladder cancer is a distinct event, with underlying molecular causes. It also identifies the TSC1 locus as a candidate for a TSG, which drives recurrence in a proportion of TCC patients. Loss of all informative markers, including those residing in the TSC1 region, spanning chromosome 9 was also linked to recurrence. PMID- 11920733 TI - Analysis of key cell-cycle checkpoint proteins in colorectal tumours. AB - Aberrations in the components of cell-cycle checkpoints are a common feature of many tumours and several have been shown to have prognostic significance in colorectal cancer. In this study, seven components of cell-cycle control [cyclin D1, retinoblastoma (pRb), p21, p27, p16, p53, and proliferating cell nuclear antigen (PCNA)] were examined in a large series of well-characterized colorectal adenocarcinomas using immunohistochemistry to ascertain co-regulation and influence on survival. The majority (92%) of the tumours had abnormal staining of > or =2 cell-cycle control factors. Expression of cyclin D1 protein was correlated with both p21 (p<0.001) and p27 (p=0.033), suggesting co-regulation of these proteins in colorectal tumours. Only cyclin D1 (p=0.048) and p53 (p=0.025) were directly associated with PCNA levels, suggesting a more important role in the proliferative capacity of tumour cells. Significant associations between cell cycle-related proteins and clinicopathological data were observed: cyclin D1 and p53 proteins were correlated with patient age (p=0.042 and p<0.001, respectively) and p53 (p=0.01) and p21 (p=0.024) proteins were associated with tumour site. Expression of cyclin D1 protein was the only protein examined that was related to improved outcome in these patients (p=0.0266), but it was not an independent predictor of survival. These results suggest that loss of control of cell-cycle checkpoints is a common occurrence in colorectal tumours and may be an important therapeutic target. PMID- 11920734 TI - 'Senescence-associated' beta-galactosidase activity in the upper gastrointestinal tract. AB - Beta-galactosidase activity at pH 6 is associated in vitro with senescence and cellular death, but in vivo data are sparse. This study undertook firstly to map 'senescence-associated' beta-galactosidase activity (SAbetaG) at pH 6 in normal epithelia and mucosae of the upper gastrointestinal tract. As escape from senescence confers a proliferative advantage, a reduction in SAbetaG activity might be predicted in neoplasia and their precursors in vivo. This prediction was tested in metaplastic, dysplastic, and neoplastic epithelium of the upper gastrointestinal tract. Histochemical staining for SAbetaG was performed at pH 6 on cryostat sections of 350 endoscopic biopsies from sites including oesophagus, stomach, and duodenum of 46 patients: 28 with Barrett's oesophagus (two with adenocarcinoma), 15 with gastric adenocarcinoma, and three with oesophageal squamous cancer. A staining score (range 0-6) was assigned to epithelial cells in all mucosae and scores were calculated for surface (luminal), intermediate, and deep (basal) layers. The strongest SAbetaG activity was in surface luminal cells of normal duodenal mucosa (mean score 3.6+/-0.5; n=19), 'specialized' Barrett's mucosa (mean 2.2+/-0.12; n=105), and intestinal metaplasia in the stomach (mean 2.4+/-0.40; n=16). Squamous epithelium was consistently negative for SAbetaG activity. Low- and high-grade Barrett's dysplasia showed no decrease in SAbetaG activity, but reduced activity was seen in gastric and oesophageal adenocarcinomas (mean 1.24+/-0.29; n=17; p=0.012). In six gastric adenocarcinomas, there was no detectable activity. Whether SAbetaG is truly a marker of cellular senescence in vivo remains to be determined. Activity is low in mucosal proliferation compartments and increases with cellular differentiation, especially in native or metaplastic intestinal mucosae. SAbetaG activity persists in dysplastic mucosae but may show some reduction or loss in adenocarcinomas (p=0.0012). Loss of SAbetaG activity is not, therefore, an early event in glandular dysplasia-neoplasia of the upper gastrointestinal tract. PMID- 11920735 TI - Malignancy-associated allelic losses on the X-chromosome in foregut but not in midgut endocrine tumours. AB - Information on genetic changes involved in the progression of gastroenteropancreatic (GEP) endocrine tumours is scanty. On the other hand, the identification of molecular markers of malignancy could be crucial for the prognostic evaluation of these neoplasms, which is hardly predictable on the basis of conventional histological criteria. An association of X-chromosome deletions with malignancy has already been found in gastric endocrine tumours. To investigate this further, a comparative loss of heterozygosity (LOH) analysis was performed on 17 pancreatic endocrine tumours (PETs) and 17 intestinal (ten ileal, six appendiceal, and one rectal) carcinoids from female patients. The relationship of X-chromosome LOH with the ploidy status of the neoplasms was also investigated. LOH was found in six of eight malignant PETs (60% of the informative markers), but was infrequent in the nine benign ones (4.5%). In contrast, although retention of heterozygosity was consistently observed in benign midgut tumours, LOH was infrequent in malignant carcinoids (15%). No correlation was found between LOH and the ploidy status. These results indicate an association between X-chromosome LOH and malignancy in foregut endocrine tumours. The lack of such an association in midgut carcinoids suggests that different molecular mechanisms are involved in the progression of these two categories of endocrine neoplasms, which are otherwise considered to be closely related. These findings emphasize the need for further molecular studies on GEP endocrine tumours, carefully subdivided according to their anatomical site of origin. PMID- 11920736 TI - Quantitative expansion of structural genomic alterations in the spectrum of neuroendocrine lung carcinomas. AB - The pathogenesis and interrelationships of neuroendocrine lung carcinomas are not well understood. Tissue macro-arrays prepared from surgical resection specimens from 35 patients with typical carcinoid (TC), six with atypical carcinoid (AC), 13 with large cell neuroendocrine carcinoma (LCNEC), and 15 with small cell lung carcinoma (SCLC) were investigated by fluorescence in situ hybridization (FISH) and immunohistochemistry. Hybridizations with locus-specific DNA probes demonstrated a high incidence of deletion for the tumour suppressor genes p53 and retinoblastoma (Rb), and for the oncogene cyclin D1, comparable in all carcinoma types. Similarly, an increase of DNA copy number for the Her-2/neu and c-myc oncogenes was noted in all neoplasms. A more detailed quantitative analysis of the results, however, demonstrated increasing numbers of cells harbouring these genomic alterations, from low-grade TC to highly malignant SCLC, with the exception of cyclin D1 deletion. Mutations of the p53 and Rb genes, as assayed by immunohistochemical studies, were observed at high incidence in high-grade carcinomas, compared with a low incidence in the low-grade carcinomas. Conversely, in all carcinoma types, neither membrane-bound Her-2/neu nor nuclear cyclin D1 was detected. It is concluded that structural genomic alterations are frequent in neuroendocrine lung carcinomas and that their occurrence may be underestimated by immunohistochemical studies alone. The quantitative expansion of the Rb, p53, c-myc, and Her-2/neu alterations towards high-grade carcinomas suggests common pathogenetic mechanisms in the spectrum of these neoplasms. PMID- 11920737 TI - Angiogenic co-operation of VEGF and stromal cell TP in endometrial carcinomas. AB - Vascular endothelial growth factor (VEGF) and thymidine phosphorylase (TP) are important angiogenic enzymes, inducing new blood vessel formation in many human malignancies. In this study, the immunohistochemical expression of the two molecules was analysed in a series of 121 endometrial carcinomas. VEGF was expressed exclusively in cancer cells, while TP expression was shown in cancer cells (TPcc) and in stromal cells (TPsc) of both fibroblastic and myometrial origin. In all cases, enzymatic detection was particularly evident at the invading tumour front. At this site, TPsc, but not VEGF, expression was associated with non-endometrioid-type carcinomas, high tumour grade, deep myometrial invasion, and advanced stage. VEGF, but not TP, expression was related to increased angiogenesis (p=0.01). Double stratification of the two factors, however, marked VEGF/TPsc co-expression as the most potent angiogenic phenotype (p=0.008), suggesting a synergistic function. Survival analysis revealed that VEGF and TPsc, whether expressed alone or in combination, define poor prognosis. In multivariate analysis, however, stage of disease (p<0.0001, t-ratio 4.4) and VEGF expression (p=0.01, t-ratio 2.4) were the most important prognostic variables. Furthermore, VEGF expression emerged as the only independent prognostic variable in stage I endometrial carcinomas (p=0.04, t-ratio 1.9). This was not shown for TP, probably because of its close association with histopathological parameters. In conclusion, VEGF is a major angiogenic factor in endometrial carcinomas and an independent prognostic factor in stage I endometrial disease. TP is not an effective contributor to the angiogenic process, but is associated with aggressive histological features. The two factors, when co-expressed, play a co-operative role in the induction of angiogenesis. PMID- 11920739 TI - FHIT expression in clear cell renal carcinomas: versatility of protein levels and correlation with survival. AB - Clear cell renal cell carcinomas (RCCs) are characterized by a deletion of chromosome 3p, which might result in the inactivation of the FHIT (fragile histidine triad) gene, a putative tumour suppressor gene. To explore the relevance of FHIT aberrations for tumour progression and prognosis in clear cell RCCs, FHIT protein expression was analysed in formalin-fixed tissue from 149 clear cell RCCs by immunohistochemistry. FHIT protein expression was found to be markedly reduced in all RCCs, when compared with adjacent non-neoplastic tubule epithelia. Although remaining below the FHIT levels of normal tubule epithelia, a significant increase of FHIT expression became evident from well (G1) to poorly (G3) differentiated clear cell RCCs (p=0.0001) and from low (pT1) to advanced (pT3) tumour stages (p=0.001). The log-rank test demonstrated a significant inverse correlation (p=0.0074) between FHIT expression and tumour aggressiveness as indicated by patient survival. Cox regression analysis revealed that FHIT expression is an independent prognostic parameter (p=0.0139) in clear cell RCCs. In conclusion, clear cell RCCs show a marked reduction of FHIT protein expression when compared with their putative cells of origin. In contrast to other tumour types, however, loss of FHIT protein expression is significantly less pronounced in poorly differentiated RCCs or advanced tumour stages. This versatility of FHIT expression during tumour progression suggests a role for reversible mechanisms of FHIT inactivation during the initiation and progression of clear cell RCCs. PMID- 11920738 TI - Involvement of the Fas/FasL pathway in the pathogenesis of germ cell tumours of the adult testis. AB - Induction of apoptosis by Fas ligand (FasL) of Fas-containing cells is a known mechanism involved in the eradication of inappropriate cells during normal development. Alterations of the Fas/FasL pathway have been found in various types of cancer, leading to circumvention of attack of the tumour by the immune system. An alternative way to circumvent eradication by induction of apoptosis is through changes in the downstream inhibitors. For example, Fas-associating phosphatase-1 (Fap-1) binds directly to the Fas receptor and results in a block of the downstream signalling. To shed more light on the role of the Fas/FasL pathway in the development of human testicular germ cell tumours of the adult testis, this study investigated the presence of Fas, FasL, Fap-1, HLA class I and II molecules, CD45 (lymphocyte marker), and CD57 [natural killer (NK) cell marker] by immunohistochemistry on frozen sections of 41 cases of seminomas, non seminomas, and spermatocytic seminomas. Every germ cell tumour was positive for Fap-1 and negative for HLA classes I and II, like their non-malignant cells of origin. The infiltrating lymphocytes, predominantly present in seminomas, showed consistently positive staining for Fas and CD45, but not for Fap-1. No Fas was found on NK cells. All seminomas and non-seminomas (except teratomas), including their precursor stages, carcinoma in situ, intratubular seminoma and intratubular non-seminoma, showed positive staining for FasL, but not for Fas. Teratoma showed no staining for FasL and was positive for Fas. In contrast, both Fas and FasL were detectable on spermatocytic seminoma. These data indicate a different regulation of the Fas/FasL system in seminoma and spermatocytic seminoma, supporting a separate pathogenesis for these germ cell-derived tumours. The presence of Fap-1 in all histological variants of germ cell tumours might be related to the consistently positive staining in cells of the germ lineage. This study indicates that production of FasL by the germ cell tumour cells might be involved in the early development of these types of adult testicular cancer by inducting apoptosis of Fas-positive, Fap-1-negative tumour-infiltrating lymphocytes. PMID- 11920740 TI - The Wnt pathway, epithelial-stromal interactions, and malignant progression in phyllodes tumours. AB - In a previous study of phyllodes tumours, it has been shown that both the stroma and the epithelium can exhibit distinct molecular changes, suggesting that both are part of the neoplastic process. In view of this finding, it was decided to study stromal-epithelial interactions in these tumours by examining the Wnt-APC beta-catenin pathway. Beta-catenin and cyclin D1 immunohistochemistry was performed on 119 phyllodes tumours. Eighty-six (72%) showed stromal nuclear beta catenin localization and in 57% the staining was moderate or strong; however, of the eight malignant tumours in the series, seven showed absent or weak nuclear staining (p<0.025). In no tumour was nuclear beta-catenin staining seen in the epithelial component. Moderate or strong stromal cyclin D1 staining correlated with nuclear stromal beta-catenin staining (p<0.05). Forty-five of the tumours, including two malignant lesions, were screened for beta-catenin exon 3 mutations using SSCP and sequencing, but none was found. Loss of heterozygosity (LOH) of the marker D5S346 was used to infer APC mutation, but only one (benign) tumour showed LOH. Wnt2 and Wnt5a mRNA was localized by in situ hybridization in 13 cases (three malignant) chosen to reflect the different beta-catenin staining patterns. There was an association between strong nuclear beta-catenin staining of stromal cells and epithelial Wnt5a expression (p<0.0015). These data suggest that stromal proliferation in benign phyllodes tumours relies on abnormalities in the Wnt pathway which result not from mutation, but from Wnt5a expression in the epithelium. In the progression to malignancy, the stromal proliferation appears to become independent of the Wnt pathway and, presumably, of the epithelial component of these tumours. PMID- 11920741 TI - APC mutations in synovial sarcoma. AB - It has previously been demonstrated that accumulated beta-catenin serves as an oncoprotein in synovial sarcoma and results in a poor overall survival rate, but the frequency of beta-catenin mutation was quite low (8.2%). The present study, using essentially the same study group of cases, screened for genetic alterations in the mutation cluster region (MCR) of the APC gene in 49 cases of synovial sarcoma. SSCP analysis followed by DNA direct sequencing revealed five missense APC mutations in four cases of synovial sarcoma (8.2%). The mutational sites comprised one case each at codons 1299 (GCT to ACT, Ala to Thr), 1412 (GGA to AGA, Gly to Arg), and 1414 (GTA to ATA, Val to Ile), in addition to one case with double point mutations at codon 1398 (AGT to AAT, Ser to Asn) and at codon 1413 (ATG to ATA, Met to Ile), together with beta-catenin mutation at codon 32 (GAC to TAC, Asp to Tyr). All four cases with APC mutations were histologically of the monophasic fibrous type and showed beta-catenin accumulation. All three cases with APC mutations available for follow-up data were long survivors. This study provides the first evidence that APC mutations also occur in the field of sarcoma, especially in synovial sarcoma. PMID- 11920742 TI - MYCN expression in human rhabdomyosarcoma cell lines and tumour samples. AB - The MYCN oncogene encodes a phosphoprotein that acts as a transcription factor and is involved in the regulation of cell proliferation and differentiation in normal as well as in cancer cells.MYCN amplification and expression have been reported in various tumours, including neuroblastoma and lung cancer, but little is known about its expression in human rhabdomyosarcoma. MYCN expression and amplification were studied in five alveolar and five embryonal rhabdomyosarcoma cell lines and in 19 tumour biopsies. All the cell lines studied expressed MYCN RNA, as demonstrated by northern blot analysis and RT-PCR, but the oncogene was amplified in only one. Similarly, MYCN protein was detected in all cell lines by western blot analysis, with higher levels of expression in alveolar than in embryonal rhabdomyosarcoma cells. RT-PCR analysis of tumour samples demonstrated 18/19 cases positive for MYCN RNA. Although MYCN expression was higher in alveolar than in embryonal rhabdomyosarcoma cell lines, no clear relationship between histology and level of MYCN expression could be established in this tumour series. These data suggest that MYCN expression is a common feature of rhabdomyosarcoma, independent of gene amplification and without a clear relationship with specific histological and clinical features. PMID- 11920743 TI - Inter-observer variation in the histopathological diagnosis of clinically suspicious pigmented skin lesions. AB - When a biopsy is taken of a suspicious pigmented skin lesion, histological examination is expected to establish the definitive diagnosis. This study evaluated the inter-observer variation of 20 pathologists in the histological diagnosis of a randomly selected set of suspicious pigmented skin lesions (PSLs), by comparing their diagnoses to a reference diagnosis. Overall sensitivity for melanoma was 87%, ranging from 55% to 100% between the observers. Sensitivity was significantly lower for thin (Breslow thickness <1 mm) than for thick melanomas (83% versus 97%, p=0.005). Overall melanoma specificity was 94%, ranging from 83% to 100% between observers. Dysplastic naevus was the most important source of false-positive diagnoses, mainly in situ melanomas. Positive and negative predictive values in the given test set were 75% and 97%, respectively. In the case of melanoma, there was quite some variation in measured Breslow thickness. This would have led to a different therapeutic approach in 12% of the readings. Some of the variation seemed to be due to a different interpretation of the presence of a co-existent naevus. In 9% (3/35) of the readings, participants did not agree on the presence of ulceration. These results reflect a tendency to overdiagnose mainly thin melanomas in general histopathological practice. They also demonstrate variation in the assessment of major prognostic factors of melanoma. PMID- 11920744 TI - Expression of the PDGF alpha-receptor 1.5 kb transcript, OCT-4, and c-KIT in human normal and malignant tissues. Implications for the early diagnosis of testicular germ cell tumours and for our understanding of regulatory mechanisms. AB - Human testicular germ cell tumours of adolescents and adults (TGCTs), including their precursor lesion carcinoma in situ (CIS), show expression of a 1.5 kb alternative transcript of the platelet-derived growth factor (PDGF) alpha receptor gene. The so-called P2 promoter involved is located in intron 12 and its activity was found to be mutually exclusive with activity of the classical promoter (P1), which encodes the full-length receptor. The presence of the 1.5 kb transcript could be a putative marker for the early molecular diagnosis of TGCTs. In order to validate the RT-PCR approach, this study shows that not more than 100 transcripts are necessary to obtain positivity in the test used; moreover, samples from TGCTs or CIS-containing tissues can be diluted many-fold before resulting in false-negative findings. This study also shows that within TGCTs, as in TGCT-derived cell lines, expression of the 1.5 kb transcript is differentiation-dependent and positively correlated with expression of the embryonic transcription factor OCT-4/POU5F1. Furthermore, the results indicate that in some non-TGCT cancers and cell lines the 1.5 kb transcript is also expressed, but without concomitant OCT-4/POU5F1 expression. The 1.5 kb transcript is also present in early B cells and derived leukaemias (B-ALL). In spite of similarities in chromosomal location, down-regulation upon differentiation of TGCTs, and PDGF alpha-receptor and c-KIT (the stem cell factor receptor) both being a tyrosine kinase receptor, no correlation was found between activity of the P2 promoter of the PDGF alpha-receptor gene and expression of c-KIT. In conclusion, the 1.5 kb transcript of the PDGF alpha-receptor is expressed in various cells and tissues, including particular blood cells. Although this may hamper the use of this transcript as a marker for malignancies in general, it does not appear to interfere with assays for the early detection of TGCTs. PMID- 11920745 TI - Atypical dose-route-dependent food effects of eplerenone in the dog: presence of food effects following intravenous dosing and lack of food effects of following oral dosing. AB - This study was conducted to investigate why a food effect was observed following an intravenous dose of eplerenone (EP) in the dog, but not following oral dosing. Three female dogs were implanted with a chronic portal vein access port and received radiolabeled EP doses orally (15 mg/kg in solution) and intravenously (7.5 mg/kg via cephalic and portal veins) under fasted and fed conditions. Mean AUC values for EP after infusion through the cephalic vein were 23.0 +/- 2.7 and 18.2 +/- 1.1 h.microg/mL under fasted and fed conditions, respectively. Corresponding values after infusion through the portal vein were 20.7 +/- 3.2 and 12.9 +/- 1.3 h.microg/mL, respectively. After oral administration, EP was absorbed 82.0 +/- 6.9 and 98.0 +/- 8.3% under fasted and fed conditions; corresponding mean AUC values were 32.0 +/- 2.0 and 30.8 +/- 3.6 h.microg/mL, respectively. The AUC value for SC-70303 acid (the open lactone form of EP) was lower under fed conditions after cephalic vein infusion, but was greater under fed conditions after portal vein infusion or oral solution administration. The hepatic first-pass effect of EP was 12.6 +/- 6.3% under fasted conditions and 27.1 +/- 6.0% under fed conditions. Pharmacokinetic analysis of EP concentrations after portal vein infusion and oral administration showed that under fed conditions the rate constants for bile excretion and for liver metabolism and urinary excretion were increased while the rate constant for elimination and/or metabolism in the gastrointestinal tract was reduced. In conclusion, the apparent lack of food effect after oral administration was observed because enhanced clearance was compensated by increased absorption. PMID- 11920746 TI - Targeted gene delivery to skin cells in vivo: a comparative study of liposomes and polymers as delivery vehicles. AB - Liposomes are microscopic lipid membrane vesicles that provide a current strategy for topical, dermal delivery of biologically active molecules. They have been successfully used for the delivery of various low and high molecular weight molecules into the skin, and as an alternative to virus-mediated delivery systems, have opened the field of dermal gene therapy. The present study was undertaken on 6-day-old rat pups to determine in vivo the efficacy of several liposome and nonliposome formulations, including phospholipid liposomes and their cationic or pegylated variants, nonionic liposomes and their cationic variant, PINC polymer (Protective, Interactive, Noncondensing polymers), and a propylene glycol:alcohol:water mixture (delivery vehicle for minoxidil) in delivering beta galactosidase and luciferase reporter genes into skin cells. Based upon our observations of the expression of beta-galactosidase and luciferase reporter genes in skin cells, we report here that nonionic liposomes are the most efficient vehicle for transdermal delivery followed by nonionic/cationic and phospholipid (pegylated) liposomes. The propylene glycol:ethanol:water mixture and the PINC polymer were relatively inefficient in the delivery of beta galactosidase or luciferase DNAs. This simple, noninvasive technique of using nonionic liposomes to deliver biomolecules provides an efficient delivery strategy for gene therapy and drug delivery to the dermal organ site. PMID- 11920747 TI - Hypericum perforatum--chemical profiling and quantitative results of St. John's Wort products by an improved high-performance liquid chromatography method. AB - The analysis of flavonoids, naphthodianthrones, and the phloroglucinol derivative hyperforin in H. perforatum is described in this article. In a 35-min HPLC run nine major compounds could be identified and baseline separated in the methanolic plant extracts. For an optimum separation the mobile phase consisted of 10 mM ammonium acetate buffer (pH 5.0) and an acetonitrile/methanol mixture; a Synergi MAX-RP 80 A column (C-12 material) was used as stationary phase. Detection was performed at 270 nm, and the identity of the compounds was confirmed in an LC-MS experiment. Commercial St. John's Wort products were analyzed and qualitative and quantitative results are discussed. PMID- 11920748 TI - Accelerated fluid bed drying using NIR monitoring and phenomenological modeling: method assessment and formulation suitability. AB - The theory behind a fluidized bed fast-drying method is investigated as a potential timesaving process, which can reduce overall drying time compared to single-temperature cycles. The method teaches that for formulations in which heat transfer dominates the drying mechanism, an increase in temperature during the evaporative phase of drying decreases overall process time by 50%, without changing the physical properties of the active ingredient. The method is also examined in terms of its potential for cycle shortening for a given formulation. A combined expression for the overall drying profile has been developed and committed to a macro that facilitates the identification of phenomenological trends, which identify whether or not a process will benefit by using fast drying. The relationship and macro also allows prediction of time frames associated with a given set of dryer and formulation parameters. PMID- 11920749 TI - Transport across 1,9-decadiene precisely mimics the chemical selectivity of the barrier domain in egg lecithin bilayers. AB - The barrier domain solubility-diffusion theory of lipid bilayer permeability relates the permeability coefficient (P(m)) to the solute's partition coefficient (PC(barrier/w)) and diffusion coefficient (D(barrier)) in the ordered chain region of the bilayer that serves as the barrier region for polar permeants. To select the best solvent to mimic the barrier domain, permeability coefficients across a layer of 1,9-decadiene were compared with permeability coefficients from bilayer transport. Rate constants for transport, k, of alpha-methyl substituted analogues of p-toluic and p-methylhippuric acid were measured across a layer of 1,9-decadiene embedded in a PTFE filter membrane placed between two aqueous solutions in side-by-side diffusion cells. Permeability coefficients (P(1,9 decadiene)) were normalized to that obtained for p-toluic acid, which was included in donor solutions. The correlation of log(P(bilayer)) versus log(P(1,9 decadiene)) was linear with a slope of 0.99 +/- 0.02 SD, indicating that 1,9 decadiene precisely mimics the egg lecithin bilayer barrier domain in its chemical selectivity. Using the decadiene membrane transport method to indirectly estimate partition coefficients for similarly sized permeants extended the range of measurable values beyond those readily attainable by the traditional shake flask method, allowing measurement of 1,9-decadiene/water PCs as low as 3 x 10( 7). PMID- 11920750 TI - A physicochemical basis for the extensive intestinal lymphatic transport of a poorly lipid soluble antimalarial, halofantrine hydrochloride, after postprandial administration to dogs. AB - The highly lipid soluble, free-base form of halofantrine (Hf base; approximately 50 mg/mL in triglyceride lipids), a highly lipophilic (calculated log P approximately 8.5) antimalarial, has recently been shown to undergo significant intestinal lymphatic transport (54% of administered dose) after postprandial administration to dogs. In contrast, the clinically available hydrochloride salt of Hf (Hf small middle dot HCl), was not considered to be a likely substrate for lymphatic transport because its solubility in long-chain triglyceride lipids is low (< 1 mg/mL). This paper reports the lymphatic transport of Hf after postprandial administration of Hf.HCl, which was surprisingly high at 47% of the administered dose, and not significantly different from that of Hf base. It was postulated that partial conversion of solubilized Hf.HCl to the highly lipid soluble Hf base within the intestinal lumen might account for the extensive lymphatic transport. However, as Hf is a tertiary amine with an expected pK(a) of > 10, at gastrointestinal pH, the fraction of Hf present as the free base form is likely to be extremely low. Physicochemical studies exploring the solubility and pK(a) of Hf suggest that Hf.HCl was extensively solubilized following fed administration. When solubilized in representative fed state mixed micellar solutions, its apparent pK(a) was 6.92 and considerably lower than anticipated for a tertiary amine. It appears that the extensive lymphatic transport of Hf observed after postprandial administration of Hf.HCl was likely to be due to the conversion of solubilized Hf.HCl to the free base. Therefore, in addition to indicators such as log P and triglyceride solubility, factors such as drug solubilization in representative fed state intestinal conditions and the possible conversion to the un-ionized form should be considered when predicting the potential lymphatic transport of salts of poorly water soluble acids and bases. PMID- 11920752 TI - New and better protocols for a short-term Caco-2 cell culture system. AB - The aim of the present study was to develop new and better protocols for a short term Caco-2 cell culture system for use in rapid screening of intestinal drug absorption. Caco-2 cells were cultured according to several protocols for short term cell culture to obtain monolayers. The effects of serum (fetal bovine serum, FBS) in the culture medium and of the period of cell culture on the barrier function and transporter activities of the monolayers were examined. The barrier function was estimated both from the transepithelial electrical resistance (TEER) and the permeability of [(14)C]mannitol. Transporter activities were monitored by measuring the permeability of [(14)C]glycylsarcosine for oligopeptide transporter (PepT1) and of rhodamine 123 for P-glycoprotein (P-gp). Caco-2 monolayers obtained by 3-day culture in the BIOCOAT HTS Caco-2 Assay System, developed by Becton Dickinson Bioscience, showed much higher permeability to hydrophilic compounds, such as mannitol, compared with those obtained by the standard 21-day culture system, due to the leaky structure of cell junctions. The newly developed 3-day protocol, which includes 10% FBS in the culture medium during the first day of culture, markedly enhanced TEER and lowered mannitol permeability of the monolayers. This protocol allowed us to better determine the rank order of permeability of compounds, giving results equivalent to those in the 21-day culture system. The longer culture period gave tighter monolayers, and the maximum value of TEER was obtained with 5 days in culture. However, after 5 days in culture, the integrity of monolayers decreased gradually. The highest activities of transporters, PepT1 and P-gp, in monolayers were obtained at days 5 or 6 of culture by the new protocol with FBS-containing medium. These results indicate that by a simple modification of the short-term culture protocol, it is possible to obtain Caco-2 monolayers with better barrier properties and higher activity of transporters that are equivalent to those found in the 21-day Caco-2 culture system. PMID- 11920751 TI - Effect of mono- and di-acylation on the ocular disposition of ganciclovir: physicochemical properties, ocular bioreversion, and antiviral activity of short chain ester prodrugs. AB - A series of short-chain carboxylic mono- and diesters of ganciclovir were synthesized in our laboratory. Physico-chemical properties, i.e., solubility (pH 4.2), partition coefficient in 1-octanol/phosphate buffer (pH 7.4), aqueous stability at various pH values, bioreversion kinetics in various ocular homogenates and effectiveness against various Herpes viruses in vitro were determined. The compounds exhibited a decrease in solubility as the ester length ascended with a corresponding increase in the octanol/buffer partition coefficient values. All of the prodrugs exhibit stability profiles typical of a carboxylic ester with maximum stability at neutral or slight acidic pH (4.0-7.0). Apparent first-order rate constants associated with prodrug to drug hydrolysis in the ocular homogenates varied depending on the size of the promoiety, lipophilicity of the compound, and the ocular tissue studied. The acetyl and butyryl mono and diesters were screened against various Herpes viruses. The monobutyrate ester of ganciclovir exhibits excellent activity against HSV-2 and VZV and provides a very high selectivity index against most of the viruses studied. PMID- 11920753 TI - Drug interactions between HIV protease inhibitors based on physiologically-based pharmacokinetic model. AB - A Physiologically-based pharmacokinetic (PB-PK) model was developed to describe the aspects of pharmacokinetic interactions between five HIV protease inhibitors (ritonavir, amprenavir, nelfinavir, saquinavir, indinavir) in rats. To increase usefulness of this BP-PK model, liver, intestinal tissue and other organ were assumed as compartments in this model. Each compartment was linked with the blood flow and the blood-to-plasma concentration ratios of those drugs, and the absorption process in the intestinal tract was presumed as a first-order kinetics. In addition, this PB-PK model incorporates two elimination processes due to hepatic and intestinal metabolism constructed by in vitro metabolic clearance rates and inhibition constants between HIV protease inhibitors. Excellent agreements were obtained between the predicted and observed concentrations of HIV protease inhibitors in rat plasma after a 20 mg/kg oral dose or co-administration of two kinds of HIV protease inhibitors (amprenavir/indinavir, nelfinavir/amprenavir, saquinavir/amprenavir, amprenavir/ritonavir, indinavir/ritonavir, nelfinavir/ritonavir, and saquinavir/ritonavir) with each 20 mg/kg oral dose. However, underestimates in the predicted plasma concentrations of saquinavir, indinavir and amprenavir were observed during the terminal phase after co-administration with ritonavir or amprenavir, suggesting that a term of other inhibitory process, such as a mechanism-based inhibition, might be incorporated into this PB-PK model. This BP PK model enables us to know useful information about pharmacokinetic interaction when HIV infected patients would receive double protease therapy. PMID- 11920754 TI - Water diffusion in hydrated crystalline and amorphous sugars monitored using H/D exchange. AB - Water interacts with pharmaceutical materials in a number of different ways. The aim of this study was to investigate if exchange experiments with D(2)O can provide useful insights into the structure of hydrated materials. Raffinose pentahydrate, trehalose dihydrate, and sucrose were used as model compounds in conjunction with their amorphous counterparts. Following exposure to D(2)O vapor, the exchange of water of hydration and/or hydroxyl groups was monitored using Raman spectroscopy. For the amorphous materials, all of the sugar hydroxyl groups were found to exchange on exposure to D(2)O, providing evidence that water has no fixed site in amorphous materials, nor is access to different parts of the molecule restricted. For raffinose pentahydrate and trehalose dihydrate, exchange of both hydrate water and hydroxyls was incomplete, suggesting that there are specific pathways for diffusion into and within the crystal structure. The results are rationalized based on the known crystal structures. Using exchange experiments to investigate hydrates thus appears to be a useful probe of structure. PMID- 11920755 TI - Characterization of carbamazepine-Gelucire 50/13 microparticles prepared by a spray-congealing process using ultrasounds. AB - In this work, the utilization of a spray-congealing technique using a new ultrasonic atomizer to prepare enhanced-release, solvent-free microspheres of carbamazepine (CBZ)-Gelucire 50/13 in different drug-to-polymer ratios was considered. Scanning electron microscopy analysis showed that it was possible to obtain spherically shaped and nonaggregated microparticles; the prevalent particle size was in the range 150-250 microm and the microspheres had a good encapsulation efficiency (> 90% in the prevalent size fraction). The in vitro dissolution tests displayed a significant increase of the CBZ dissolution rate from microspheres compared with pure drug and to drug-Gelucire 50/13 physical mixture. Differential scanning calorimetry, hot stage microscopy, X-ray powder diffractometry, and diffuse reflectance Fourier transform infrared spectroscopy demonstrated phase stability of the original polymorph of CBZ in all the systems; moreover, no interactions between the drug and Gelucire 50/13 were found. The results of this study suggested that the spray-congealing technique using the ultrasonic atomizer could be considered as a new and interesting method to enhance the dissolution rate of a poorly water-soluble drug as CBZ. PMID- 11920756 TI - Solid-state properties of creatine monohydrate. AB - Creatine monohydrate (CM) is a nutritional supplement and an ergogenic aid for athletes. It appears to increase lean body mass, high-intensity power output and strength in healthy humans. The crystal structure of creatine monohydrate has previously been reported. However, little information is available on its solid state properties. In this investigation, creatine monohydrate was subjected to Thermal Analyses, Karl-Fisccher Titrimetry (KFT), Scanning Electron Microscopy (SEM), and Variable Temperature X-ray Powder Diffractometry (VTXRD) to characterize its solid-state properties. The results of this study suggested that commercially available creatine monohydrate dehydrates at about 97-125 degrees C. A phase transition after dehydration was confirmed by X-ray diffraction studies. This dehydrated phase at a temperature above 230 degrees C undergoes intramolecular cyclization with a loss of an additional mole of water to form creatinine. Creatinine finally melts with decomposition at about 290 degrees C. VTXRD, confirmed that the above solid-state thermal transformation was kinetically driven, and occurred within a narrow temperature range. Mass Spectrometric (MS) studies further indicated a possible dimerization of creatinine formed during the solid-state transformation. PMID- 11920757 TI - Tacrolimus is a class II low-solubility high-permeability drug: the effect of P glycoprotein efflux on regional permeability of tacrolimus in rats. AB - The objective of this study is to investigate the role of P-glycoprotein (P-gp), a membrane efflux pump associated with multidrug resistance (MDR) and a known substrate for tacrolimus, in determining the regional intestinal permeability of tacrolimus in rats. Thus, isolated segments of rat jejunum, ileum, or colon were perfused with tacrolimus solutions containing polyethoxylated hydrogenated castor oil 60 surfactant, and with or without verapamil, a P-gp substrate used to reverse the MDR phenotype. The results indicated that the intrinsic permeability of tacrolimus in the jejunum, calculated on the basis of the concentration of non micellized free tacrolimus, was quite high ( approximately 1.4 x 10(-4) cm/s). The apparent permeability (P(app)) in the jejunum was unaffected by the presence of verapamil; however, the P(app) in the ileum and the colon increased significantly in the presence of verapamil and were similar to the values observed in the jejunum. The results suggest that systemic absorption of tacrolimus from the gastrointestinal tract could be significantly affected by P gp efflux mechanisms. It is also possible that differences in P-gp function at various intestinal sites in a subject or at a given intestinal site in various subjects could lead to large intra- and interindividual variability in bioavailability of tacrolimus following oral administration. PMID- 11920758 TI - Mechanisms of cytoprotective effect of amino acids on local toxicity caused by sodium laurate, a drug absorption enhancer, in intestinal epithelium. AB - Several amino acids, including L-glutamine (L-Gln), were found to protect the intestinal epithelial cells from the local toxicity caused by a drug absorption enhancer, sodium laurate (C12), in our previous study. To develop more efficient and safer formulations for enhancing drug absorption, the mechanisms of cytoprotection by amino acids were studied using rats and Caco-2 cells. Four amino acids, including L-Gln, could generally maintain the absorption-promoting action of C12, although taurine tended to attenuate it. Three amino acids, except for L-Gln, significantly suppressed the decrease in the transepithelial electrical resistance caused by C12. Quercetin, an inhibitor for biosynthesis of heat shock protein 70 (HSP70), masked only the protective effect of L-Gln in both rat large intestine and Caco-2 cells. Western blot analysis indicated clearly that HSP70 is induced extensively only by the addition of L-Gln in both rat large intestinal cells and Caco-2 cells. C12 was found to increase the intracellular concentration of Ca(2+) ([Ca(2+)](i)) remarkably, and amino acids, especially L arginine, L-methionine, and taurine, significantly attenuated the increase in [Ca(2+)](i) caused by C12. Furthermore, although C12 stimulated the release of histamine, an inflammatory mediator, from rat large-intestinal tissue, amino acids were also found to suppress the release of histamine enhanced by C12. The results in the present study showed that an induction of HSP70, a decrease in [Ca(2+)](i) elevated by C12, and a suppression of histamine release stimulated by C12 should be involved in the mechanisms behind the cytoprotective action of amino acids against the local toxicity caused by C12. PMID- 11920759 TI - A theoretical analysis of permeation of small hydrophobic solutes across the stratum corneum based on Scaled Particle Theory. AB - The barrier properties of skin originate from its lipid bilayers whose ordered structure retards solute transport. The objective of this study is to develop a mathematical model that can predict skin permeability to small (MW < 500 Da), hydrophobic solutes based on the fundamental transport properties of skin lipid bilayers. We developed a mathematical model to predict two major transport properties (i.e., partition and diffusion coefficients) using important structural properties of lipid bilayers and molecular properties of the solute. The predictions are based on Scaled Particle Theory that calculates these properties using statistical mechanics of lipid chains. The calculations predict that solute partition coefficients in lipid bilayers are of the same order as those measured in isotropic solvents, such as octanol (K(o/w)). On the other hand, solute diffusion coefficients decrease exponentially with solute cross sectional area. The resulting equation to predict skin permeability is given by P = 5.6 x 10(-6)K(0.7)(o/w)exp(-0.46r(2)(A1)--where r is solute molecular radius in Angstroms (A) and P is in cm/s. The predicted skin permeabilities compare well with the experimental data. PMID- 11920760 TI - Porous resins as a cavitation enhancer for low-frequency sonophoresis. AB - The application of low-frequency ultrasound enhances drug transport through the skin, a phenomenon referred to as low-frequency sonophoresis. This enhancement is mediated through cavitation, the formation and collapse of gaseous bubbles. We hypothesized that the efficacy of low-frequency sonophoresis can be significantly enhanced by provision of nuclei for cavitation. In this study, we used two porous resins, Diaion HP20 and Diaion HP2MG (2MG), as cavitation nuclei. We measured the effect of these resins on cavitation using pitting of aluminum foil. 2MG showed a higher efficacy in enhancing cavitation compared with Diaion HP20. 2MG was also effective in enhancing transdermal mannitol transport. These results confirm that the addition of cavitation nuclei such as porous resins further increases the effect of low-frequency ultrasound on skin permeability. PMID- 11920761 TI - Anticoagulant activity of heparin following oral administration of heparin-loaded microparticles in rabbits. AB - Heparin-loaded microparticles, prepared according to the double emulsion method with biodegradable (PCL and PLGA) and nonbiodegradable (Eudragit RS and RL) polymers used alone or in combination, with or without gelatin, were characterized in vitro and in vivo after oral administration in rabbits. The entrapment efficiency and the release of heparin were determined by a colorimetric method with Azure II. The antifactor Xa activity of heparin released in vitro after 24 h was assessed. After oral administration of heparin-loaded microparticles in rabbits, the time course of modification of the clotting time estimated by the activated partial thromboplastin time (APTT) was followed over 24 h. Microparticles with a size ranging from 80 to 280 microm were obtained. Heparin entrapment efficiency as well as heparin release depended on both the nature of the polymers and the presence of gelatin. The Eudragit polymers increased the drug loading but slowed down the heparin release, whereas gelatin accelerated the release. No change in clotting time was observed after oral administration of gelatin microparticles. Heparin-loaded microparticles prepared with blends of PLGA and Eudragit displayed a prolonged duration of action characterized by a twofold increase in APTT and a enhancement of absorption. This study demonstrated the feasibility of encapsulating heparin within polymeric particles, and the significant increase in APTT confirmed the oral absorption of heparin released from polymeric microparticles. PMID- 11920762 TI - Muscle distribution of the neuromuscular blocker gallamine using microdialysis. AB - Measurement of drug concentrations in target tissue has the potential to provide insight into the pharmacokinetics and pharmacodynamics of a drug. In this study, the distribution of the neuromuscular blocker, gallamine, into muscle tissue was investigated in urethane-anesthetized rats after an intravenous bolus dose (6 mg/kg). Microdialysis sampling was used to continuously determine gallamine concentrations in muscle interstitial fluid (MIF). In vivo microdialysis recovery of gallamine was determined as the relative loss of gallamine from the perfusate into muscle tissue after perfusion with gallamine (2 microg/mL). Recovery was determined in each rat before the pharmacokinetic studies. Terminal muscle sampling followed by homogenization was also performed to examine gallamine distribution within muscle tissue. All samples were assayed for gallamine using a validated high-performance liquid chromatography assay. Gallamine was rapidly distributed into MIF with a MIF-plasma partition coefficient of 0.9 +/- 0.1 (n = 6). By contrast, the estimated gallamine concentration in muscle tissue homogenate was only 23 +/- 5% (n = 5) of the concentration in MIF as estimated by microdialysis sampling at the terminal sampling time. These findings suggest that gallamine is not distributed uniformly within muscle but selectively distributes into MIF. Simulations using a hybrid physiologically based pharmacokinetic model which describes uptake of drug only into the interstitial space showed good agreement between predicted and observed concentration data obtained from microdialysis sampling, supporting the findings that gallamine selectively distributes into MIF. These studies demonstrate microdialysis combined with conventional terminal tissue sampling provides valuable information on intra tissue drug distribution. PMID- 11920763 TI - A new method for characterizing the release of drugs from tablets in low liquid surroundings. AB - The purpose of this article is to introduce a method capable of determining early drug dissolution in small amounts of liquid. The method is based on the measurement of the alternating ionic current through a cell containing the dissolution medium and the substance to be dissolved. Both the initial and more prolonged absorption of liquid into tablets can also be determined by using the same technique. The method has been tested on two tablet formulations containing agglomerated micronized cellulose and NaCl as a model drug. Release of NaCl was delayed from both formulations; the extent of the delay was strongly formulation dependent only when the surrounding liquid was in short supply. This finding shows that new drug dissolution phenomena may be encountered in small liquid volumes; these phenomena would not have been seen with the large volume methods normally used in in vitro dissolution tests. Hence, for formulations intended for sublingual, buccal, or rectal administration, i.e., in areas where liquid is scarce, in vitro dissolution tests should be performed in small volumes of dissolution medium. PMID- 11920764 TI - Testosterone 17beta-N,N-dimethylglycinate hydrochloride: A prodrug with a potential for nasal delivery of testosterone. AB - The purpose of this study was to examine the potential of the nasal route for the systemic delivery of the poorly water-soluble drug testosterone (TS) using a water-soluble prodrug, TS 17beta-N,N-dimethylglycinate hydrochloride. The physicochemical properties of the prodrug, in vitro hydrolysis in human liver homogenate, and in vivo nasal and intravenous experiments were performed in rats. The aqueous solubility of the prodrug was more than 100 mg/mL, compared with 0.01 mg/mL for TS, and its log partition coefficient between 0.05 M, phosphate buffer (pH 6) and octanol was 2.4. The prodrug was found to generate TS in 33% human liver homogenate and was absorbed from the nasal cavity rapidly and quantitatively. The bioavailabilities of both the prodrug and TS after nasal administration of the prodrug were similar to that after equivalent intravenous doses. These studies in rats suggest that this water-soluble prodrug of TS may have therapeutic utility for the management of TS deficiency. PMID- 11920765 TI - Poly(lactide-co-glycolide) microspheres for the controlled release of oligonucleotide/polyethylenimine complexes. AB - In this article, microspheres able to induce the controlled release of oligonucleotide/polyethylenimine complexes are proposed. A model oligonucleotide (the oligothymidilate pdT16) was encapsulated within poly(lactide-co-glycolide) microspheres alone or associated with polyethylenimine (PEI) at different nitrogen/phospate (N/P) ratios. Microspheres were prepared by the multiple emulsion-solvent evaporation technique and characterized for morphology, diameter, encapsulation efficiency, and release kinetics. The introduction of PEI in the internal aqueous phase resulted in the formation of a soluble complex with pdT16 and in a strong increase of the oligonucleotide encapsulation efficiency. PEI affected microsphere morphology inducing the formation of very porous particles yielding to an accelerated release of pdT16. When incubated with HeLa cells, microspheres encapsulating pdT16/PEI complexes allowed both a reduction of the complex toxicity and an improvement of the intracellular penetration of the released oligonucleotide. We conclude that biodegradable microspheres encapsulating oligonucleotides/PEI complexes have a great potential as controlled release system because they allow the sustained release of an oligonucleotide carrier that crosses biological membranes and locates in nucleus. PMID- 11920766 TI - Hard to swallow dry: kinetics and mechanism of the anhydrous thermal decomposition of acetylsalicylic acid. AB - The methods of thermal analysis and mass spectrometry have been used to study the kinetics and mechanism of the anhydrous thermal decomposition of acetylsalicylic acid. Both thermogravimetric analysis (TGA) and differential scanning calorimetry (DSC) show that decomposition occurs in two steps. Mass-spectrometric analysis of the residue left after the first decomposition step (approximately equal to 60% mass loss) suggests that in the condensed phase, acetylsalicylic acid decomposes by first forming linear oligomers that are further converted into cyclic oligomers. Model-free isoconversional kinetic analysis of TGA traces has been used to determine global activation energies as a function of the extent of reaction. This method of analysis has also been used to make kinetic predictions of shelf life at ambient temperatures (20-40 degrees C) under anhydrous conditions for acetylsalicylic acid. Our estimate of a shelf life of 876 days (approximately equal to 2.4 years) for 5% decomposition at 30 degrees C is in good agreement with shelf lives of 2-3 years that are stamped on over-the-counter aspirin bottles. Hence, this approach can be used to systematically study the factors that determine the decomposition kinetics of aspirin and may be used for express screening of pharmaceuticals in order to identify those with desirable thermal stabilities. PMID- 11920767 TI - 17beta-estradiol matrixpatch removal and reapplication in postmenopausal women: experimental results. AB - This study compares the pharmacokinetic performance of a matrix system for transdermal 17-beta-estradiol (E(2)) delivery using multiple consecutive dosing with a first application in postmenopausal women undergoing hormone replacement therapy. A clinical study (SI) was conducted over a treatment period of 11 days in 16 postmenopausal women receiving three consecutively applied matrix patches for the delivery of E(2). The first patch was worn for 4 days, the second for 3 days, and the third patch for 4 days. The E(2) plasma profiles determined during the third application were compared with results obtained by a published clinical study (SII) using the same patch in the same group of postmenopausal women without E(2) pretreatment. Additionally, the 24 h plasma profiles of E(2) and estrone were determined before and on day 4 during patch application of the third patch. Comparison of the mean pharmacokinetic parameters from the two studies showed no significant difference in E(2) plasma levels: AUC(0-->96h) [pg/mL h] SI: 4342 +/- 1513 and SII: 4512 +/- 1229; C(max)[pg/mL] SI: 51.3 +/- 28.8 and SII: 54.2 +/- 22.3; C(average) [pg/mL] SI: 45.0 +/- 13.2 and SII: 47.0 +/- 9.4; C(min) [pg/mL] SI: 31.4 +/- 5.9 and SII: 32.2 +/- 8.1. Over 96 h, fluctuation, f, defined as (C(max) - C(min)) / C(average), was 0.44 in SI and 0.47 in SII. Individual comparison of E(2)-C(max), -AUC, and -C(min) revealed that more than 87.5% of all patients showed a variation between SI and SII of less than 10%. The mean of the individual AUC(0-->96h) variation between the first and the third patch was only 4.7%. There was no significant drop in E(2) plasma values after patch removal and reapplication, and accumulation of E(2) did not occur after several patches were applied consecutively. Plasma E(2) showed a circadian rhythm that was lower in the morning and higher in the evening. No circadian rhythm was observed in untreated basal plasma E(2) in the group of postmenopausal women. The transdermal matrix system yielded sustained E(2) plasma levels in postmenopausal women in the initial application period. In long-term dosing there was no accumulation of E(2) in plasma and no significant drop after patch removal. It is presently not known why the circadian variation in the experimentally obtained E(2) plasma values exists. PMID- 11920768 TI - 17beta-estradiol matrixpatch removal and reapplication in postmenopausal women: theoretical predictions with an oscillating diffusion coefficient model. AB - The pharmacokinetic performance of a matrix system for transdermal beta-estradiol (E(2)) delivery after multiple consecutive dosing in postmenopausal women undergoing hormone replacement therapy was investigated. The E(2) plasma profiles determined during the third application in 16 postmenopausal women were compared with results obtained in a published clinical study using the same patch in 24 postmenopausal women without E(2) pretreatment; they were compared with a theoretical diffusion/pharmacokinetic model. A conventional theoretical model with constant model parameter (CPM) obtained from in vitro mass balance experiments in a Franz cell type set up described successfully the transdermal E(2) bioavailability parameter AUC(0-96h) (4341.9 +/- 1513.1; calculated 4250.8) and C(average) (45.0 +/- 13.2; calculated 41.2). Also, experimentally, there was no significant drop in E(2) plasma values after patch removal and reapplication; this was corroborated by calculations. Accumulation of E(2) did not occur when several patches were applied consecutively over a period of 3 weeks. Steady state was achieved following application of the first patch. However, the differences between recorded E(2) plasma profiles and theoretical results detected at specific measurement points cannot be explained by the CPM model. Experimentally obtained plasma profiles were always lower in the morning and higher in the evening than predicted on the basis of the model. Measurements of in vivo skin temperature in the postmenopausal women showed oscillating temperature profiles in the form of a cosinor function: The temperature mesor of untreated postmenopausal women was 34.8 degrees C with an acrophase at 17.0 o'clock (95% CI: 14.30-19.30) and an amplitude of +/- 0.4 degrees C (p = 0.1). During the application of the patch the average temperature next to a patch rose 0.3 degrees C, which was statistically significant (p = 0.1). In the skin under the application of the matrix patch a mesor temperature was detected as 35.6 degrees C with an amplitude of +/- 0.5 degrees C with an acrophase at 17.51 o'clock (95% CI: 14.30-21.00) (p = 0.05). The temperature period was 24 h for all measurements and the maximum temperature was observed at about 16.30 h, and a minimum at about 5.00 h. A linear dependency was detected in in vitro experiments between the log of E(2) permeability and the temperature for stripped skin, epidermis/dermis layer, as well as for the matrix. Modeling of E(2) plasma profiles with oscillating diffusion coefficients (ODM1) with a sine wave function results in this equation: D(1) = D(0x) + Da(x).sin(k.t). D(0x) is the diffusion coefficient determined at 35.6 degrees C, k is 1/24 h, D(a) is the diffusion coefficient of the temperature amplitude, h is hour, and x stands for the respective diffusion layer. It was shown that the experimental E(2) plasma profile variations are more pronounced than can simply be explained by skin temperature variations alone (ODM1 model). A simplex fit with an oscillating diffusion coefficient in the form of a sine wave function for the stratum corneum (ODM2 model) resulted in a temperature amplitude of 1.1 degrees C, about twice as high as was determined in the in vivo measurements (ODM2 model). Therefore, other circadian parameterlike blood flow might superimpose the temperature profile. The improvement in data analysis by incorporating oscillating diffusion coefficients (ODM1 and 2) over CPM was judged from a comparison of experimental data with the calculated plasma profiles with the AIC, Akaikes model selection criterion, which allows ranking between models because it is independent of the scaling of the data points. ODM1 and ODM2 improved the data analysis over CPM by allowing better calculation of experimental C(max), t(max), the time to reach to C(max), and the fluctuation, f. No difference between CPM, ODM1, or ODM2 was found for the bioavailability parameter C(average) and AUC(0-96h). PMID- 11920769 TI - Release mechanisms from gentamicin loaded poly(lactic-co-glycolic acid) (PLGA) microparticles. AB - To provide local gentamicin delivery for 1 week based on a biodegradable system, poly(lactic-co-glycolic acid) (PLGA) microparticles were developed utilizing a 50/50 blend of Resomer RG 502H, an uncapped variety of 13.5 kD, and Resomer RG 503, an endcapped polymer of 36.2 kD. The liberation mechanism was investigated by analysis of morphological changes and thermal analysis focusing on the polymer glass transition temperature (T(g)) and the mechanical properties. The release of gentamicin was related to a structural breakdown of the particles reaching a critical molecular weight. A T(g) of < 37 degrees C in the hydrated state was not indicative of collapse and agglomeration of the particles because the mechanical strength of the polymer structures in the rubbery state may still render sufficient support. As the gap between incubation temperature and T(g) widened, the mechanical stability of the PLGA microparticles decreased and became decisive. Particles prepared with RG 502H show a lower ability to bear mechanical stress than RG 503 and 50/50 RG 502H/RG 503 microparticles. PMID- 11920770 TI - Interactions of cyclosporines with lipid membranes as studied by solid-state nuclear magnetic resonance spectroscopy and high-sensitivity titration calorimetry. AB - Cyclosporin A (CyA) interacts with lipid membranes. Binding reaction and membrane location of CyA and analogs were examined with 2H-NMR, high-sensitivity isothermal titration calorimetry (ITC), and CD spectroscopy. Effects of CyA and charged analogs on the phosphocholine head group and on the membrane interior were investigated using selectively deuterated phospholipids. Incorporation of cyclosporin generated small disordering of the lipid acyl chains. Binding of CyA and neutral and positively charged analogs to lipid membranes showed endothermic heats of reaction between + 5.9 and + 11.3 kcal/mol, whereas enthalpy of binding was close to zero for the negatively charged derivative. Binding constants of cyclosporines to liposomal membranes were in the range of K(P) = 1650-5560 M(- 1) depending on the cholesterol content. (2)H-NMR provides evidence that CyA is essentially located in the interior of the bilayer membrane. For the charged analogs an additional interaction occurs at the head group level, placing the polar groups of these CyA analogs in the vicinity of the phosphocholine dipoles. The association of CyA and its analogs is accompanied by a positive enthalpy change, which is overcompensated by positive entropy changes. Binding of CyA to lipid membranes thus follows the classical hydrophobic effect, which is in contrast to many other peptide-lipid binding reactions. PMID- 11920771 TI - Effect of oral ketoconazole on first-pass effect of nifedipine after oral administration in dogs. AB - The long-term oral ketoconazole (KTZ) treatment extensively inhibits hepatic CYP3A activity. We investigated the effect of the KTZ treatment on hepatic and intestinal extraction of nifedipine (NIF) using beagle dogs. Four dogs were given orally KTZ for 20 days (200 mg, bid). NIF was administered either intravenously (0.5 mg/kg) or orally (20 mg) 10 and 20 days before the KTZ treatment and 10 and 20 days after start of KTZ treatment. CLtot of NIF after intravenous administration decreased to about 50% during the KTZ treatment. C(max) and AUC after oral administration increased to 2.5-fold and fourfold, respectively, by the KTZ treatment. The hepatic extraction ratio of NIF decreased to about a half by KTZ. A significant decrease in intestinal extraction ratio was not observed. In conclusion, the KTZ treatment inhibits hepatic extraction more profoundly than intestinal extraction of NIF. Therefore, inhibition of hepatic extraction of NIF by the KTZ treatment mainly results in substantial increase in systemic bioavailability in dogs. Because KTZ inhibits human CYP3A activities similar to canine CYP3A activities, the long-term oral KTZ treatment may dramatically increase bioavailability of NIF or other CYP3A substrates in humans. PMID- 11920772 TI - Thermodynamic origin of the solubility profile of drugs showing one or two maxima against the polarity of aqueous and nonaqueous mixtures: niflumic acid and caffeine. AB - The purpose of this work was to investigate the origin of the different solubility profiles of drugs against the polarity of solvent mixtures with a common cosolvent. Niflumic acid and caffeine where chosen as model drugs. The solubilities were measured at five or six temperatures in aqueous (ethanol-water) and nonaqueous (ethyl acetate-ethanol) mixtures. The enthalpies of solution were obtained at the harmonic mean of the experimental temperature. Solid phase changes were analyzed using differential scanning calorimetry and thermomicroscopy. A single solubility maximum was obtained for niflumic acid against the solubility parameter of both mixtures that is not related to solid phase changes. In contrast, caffeine displays two maxima and anhydrous-hydrate transition occurs at the solubility peak in the amphiprotic mixture. The apparent enthalpies of solution of both drugs show endothermic maxima against solvent composition that are related to hydrophobic hydration. A general explanation for the cosolvent action in aqueous mixtures is proposed. The dominant mechanism shifts from entropy to enthalpy at a certain cosolvent ratio dependent on the hydrophobicity and the solubility parameter of the drug. Niflumic acid and caffeine show enthalpy-entropy compensation in ethanol-water, and this relationship is demonstrated for the first time in nonaqueous mixtures. The results support that enthalpy-entropy compensation is a general effect for the solubility of drugs in solvent mixtures. The shape of the solubility curves is correlated with the compensation plots. The solubility peaks separate different enthalpy-entropy relationships that also differentiate the solubility behavior of the hydrate and the anhydrous forms of caffeine. PMID- 11920773 TI - An automated workstation for forced degradation of active pharmaceutical ingredients. AB - This article describes a system capable of performing and analysing multiple degradation experiments on drug substances as part of the process of developing stability indicating separations methods. Qualitative data are generated on the significant primary degradation processes of the drug of interest. Ten samples are refluxed with stirring in a single heating block. The robot arm is equipped with a sampling device capable of removing aliquots, during the reflux experiment, and transferring them to an HPLC injector. On-line analysis using fast HPLC with diode array and electrospray mass spectrometric detection allows identification of degradants. The methods described offer a significant time savings compared to the previously applied manual approach, and also provide data from multiple time points for each reaction. This increased knowledge about the progress of the reaction aids us in focussing efficiently on the primary degradation processes. PMID- 11920774 TI - Estimating degradation in real time and accelerated stability tests with random lot-to-lot variation: a simulation study. AB - The effect of different lot-to-lot variability levels on the prediction of stability are studied based on two statistical models for estimating degradation in real time and accelerated stability tests. Lot-to-lot variability is considered as random in both models, and is attributed to two sources-variability at time zero, and variability of degradation rate. Real-time stability tests are modeled as a function of time while accelerated stability tests as a function of time and temperatures. Several data sets were simulated, and a maximum likelihood approach was used for estimation. The 95% confidence intervals for the degradation rate depend on the amount of lot-to-lot variability. When lot-to-lot degradation rate variability is relatively large (CV > or = 8%) the estimated confidence intervals do not represent the trend for individual lots. In such cases it is recommended to analyze each lot individually. PMID- 11920776 TI - Our operative approach to cancer has been modified by scientific innovations: is this true for gastrointestinal cancer? PMID- 11920777 TI - Validating the performance of the mammary sentinel lymph node team. AB - BACKGROUND AND OBJECTIVES: The mammary sentinel lymph node (SLN) procedure has the potential to improve the accuracy and lower the morbidity of axillary staging in breast cancer patients, but results are closely linked to experience and can vary widely between institutions. Standardized performance measures need to be established in order to optimize the transition to SLN biopsy only. METHODS: Performance data were prospectively collected for the first 156 mammary SLN procedures performed by three surgeons in our institution. RESULTS: Seventy-five cases were required to achieve an SLN visualization rate of > 80% on preoperative lymphoscintigraphy. The SLN visualization rate was 90% for the last 52 cases. Two surgeons required 25 cases before consistently achieving a > or = 90% SLN identification rate in the operating room and one required 15 cases. The metastasis detection rate increased from 22% for the first 52 cases to 31% for the last 52 cases. The false negative rate for the procedure was 5%. CONCLUSIONS: The following performance criteria and benchmarks are suggested for validating the performance of the SLN team: (1) SLN visualization rate on preoperative lymphoscintigraphy > or = 80%, (2) SLN identification rate in the operating room > or = 90%, (3) False negative rate for the procedure 5%. Thirty procedures per surgeon were sufficient to achieve these benchmarks in our group. PMID- 11920778 TI - Prognostic value of c-erbB2 expression in breast cancer. AB - BACKGROUND AND OBJECTIVES: An overexpression of c-erbB2 has been reported to be associated with a poor clinical outcome in breast cancer, however, its prognostic value remains controversial especially in patients with node negative breast cancer, and regarding the estrogen receptor (ER) status. METHODS: Immunohistochemical staining for c-erbB2 was performed on the primary breast cancer from 698 Japanese patients with a mean follow-up duration of 54 months. RESULTS: The c-erbB2 expression was positive in 120 (17.2%) of 698 cases, which inversely correlated with the ER status. Both univariate and multivariate analyses indicated the c-erbB2 expression to be a significant prognostic factor for the disease-free survival (DFS) and overall survival (OS), while the same effect was also seen in the patient groups with node negative as well as node positive breast cancer. A univariate analysis also indicated a subgroup with the positive c-erbB2 and negative ER to have both a worse DFS and OS than the other subgroups. The patients with positive c-erbB2 had a significantly worse DFS and OS than the patients with negative c-erbB2 in all patient groups stratified according to the adjuvant therapies, while the prognostic significance of c-erbB2 on DFS was also found in the patients with the node negative breast cancer who received adjuvant tamoxifen alone. CONCLUSIONS: The c-erbB2 expression is an independent significant factor for breast cancer and the prognostic significance remains in the node negative as well as node positive breast cancer, while the same effect was also found in all subgroups stratified according to the adjuvant therapies. In addition, the combination of c-erbB2 and ER made it possible to identify the subgroup with the worst clinical outcome. PMID- 11920779 TI - Clinical implications of lymph node micrometastasis in patients with histologically node-negative (pN0) esophageal carcinoma. AB - BACKGROUND AND OBJECTIVES: Lymph node micrometastasis is frequently detected in resected specimens of esophageal squamous cell carcinoma (ESCC). The goal of this study was to evaluate the clinical implication of micrometastasis in patients with lymph node-negative (pN0) disease. METHODS: We examined the presence of micrometastasis in 2,511 lymph nodes from 53 patients with pN0 disease who underwent curative esophagectomy. All lymph nodes and the primary tumors were immunostained using an anticytokeratin antibody cocktail (AE1/AE3). RESULTS: Micrometastasis was detected in 18 lymph nodes (0.72%) from 14 patients (26.4%). Detection of micrometastasis was not associated with the depth of invasion or the differentiation or lymphatic invasion. Lymph nodes containing micrometastases were widely distributed, but the most frequently involved nodes were located along the lesser curvature of the stomach. Four patients with micrometastasis (29%) and the only two patients without micrometastasis (5%) had recurrence as lymph node metastases (P = 0.036). There were no significant differences in recurrence-free survival and disease-specific survival between patients with micrometastasis and patients without micrometastasis. CONCLUSIONS: These results show that micrometastasis might increase the risk of lymph node recurrence, but does not influence the survival of patients with pN0 ESCC. PMID- 11920780 TI - Expression of CD44v6 in advanced gastric cancer and its relationship to hematogenous metastasis and long-term prognosis. AB - BACKGROUND AND OBJECTIVES: Variants of CD44 have been proposed to be important in promoting tumor progression and metastasis. We attempted to determine the expression of CD44v6 product in advanced gastric cancer and to evaluate its prognostic value. METHODS: The expression of CD44v6 was analyzed immunohistochemically in advanced gastric cancers using monoclonal antibody, 44 2V. We investigated the relationship between CD44v6 expression and prognosis in 201 gastric cancer patients. RESULTS: Ninety-five (47.3%) of 201 cancer tissues expressed CD44v6. The expression of CD44v6 protein was significantly higher in differentiated, adenocarcinoma than in diffuse type carcinoma. The CD44v6 positive cancers were more frequently associated with hematogenous metastasis. There was no significant correlation between CD44v6 immunoreactivity, and prognosis among the combined cases. Among patients with differentiated adenocarcinoma, however, the prognosis was significantly poorer in patients with CD44v6-positive tumors than in those with CD44v6-negative tumors. CONCLUSIONS: CD44v6 protein may have an important role in hematogenous metastasis, and may be a biologic marker of prognostic significance in differentiated type gastric cancers. PMID- 11920781 TI - Apoptosis and its correlation with proliferative activity in rectal cancer. AB - BACKGROUND AND OBJECTIVES: Alterations in the normal control of apoptosis and cell proliferation are important factors in multistep colorectal carcinogenesis. The aim of this study was to determine the frequency of apoptosis and cell proliferation in rectal cancers and to examine their relationship to clinicopathological variables and expression of bcl-2 and p53. METHODS: Terminal deoxynucleotidyl transferase (TdT)-mediated dUTP nick end labeling (TUNEL) staining and immunohistochemical staining for Ki-67, bcl-2, and p53 were performed on paraffin-embedded tissue samples of 57 rectal cancers. RESULTS: There was a positive linear correlation between apoptotic index (AI) and proliferative index (PI) (gamma = 0.276, P = 0.038). Both apoptosis and cell proliferation were more frequently found in rectal cancers with lymph node metastasis (P = 0.045 and 0.010, respectively). However, the ratio of AI and PI was not different by nodal status. There was no association between Dukes stage and AI or PI. The frequency of apoptosis was inversely related to the expression of bcl-2, but was not related to the p53 status of rectal cancer. There were no association between cell proliferation and the expression of bcl-2 or p53. CONCLUSIONS: Our results suggest that the susceptibility to apoptosis in rectal cancer is clearly related to the proliferative activity and high turnover rate of tumor cells may contribute to lymph node metastasis. PMID- 11920782 TI - Malignant transformation of "benign" cystic mesothelioma of the peritoneum. AB - BACKGROUND AND OBJECTIVES: Peritoneal mesothelioma is being diagnosed with greater accuracy as a result of immunocytochemical analysis. The histological type of peritoneal mesothelioma has a great influence on the natural history of the disease. Benign cystic mesothelioma is a definite clinical entity; however, the absence of a uniform approach to treatment and a lack of long-term follow-up of patients seriously hinders an accurate assessment of the disease process. METHODS: The medical history of a patient with a diagnosis of benign peritoneal cystic mesothelioma followed for 10 years is presented. The medical literature of this disease is reviewed. RESULTS: The patient was a 36-year-old woman initially diagnosed with benign cystic peritoneal mesothelioma; however, after six surgical procedures with the aim of reducing the volume of intraperitoneal fluid and cysts, the disease developed into an aggressive, diffuse malignant mesothelioma. Involvement of abdominal incisions, invasion of lymph nodes, and invasion of the viscera occurred. CONCLUSIONS: Peritoneal cystic mesothelioma is a disease in need of careful longitudinal studies in order to better define the clinical course of these patients. This case report along with a literature review suggests caution in the assessment of cystic mesothelioma as a benign process. This patient had a clear malignant transformation of benign cystic mesothelioma to an invasive and potentially lethal process. PMID- 11920783 TI - Isolated limb perfusion for melanoma. PMID- 11920784 TI - Simple technique to approach bleeding of the pancreatic stump after pancreaticoduodenectomy. PMID- 11920785 TI - Laparoscopic-assisted sigmoidectomy with lymph node dissection via minilaparotomy. PMID- 11920787 TI - Pharmacokinetics of acyclovir in immunocompromized children with leukopenia and mucositis after chemotherapy: can intravenous acyclovir be substituted by oral valacyclovir? AB - BACKGROUND: The efficacy of oral acyclovir, a purine nucleoside analogue with activity against human herpes viruses, is limited as a result of its low bioavailability. Valacyclovir, the L-valyl ester of acyclovir, has been developed as a pro-drug to improve the bioavailability. The aim of the present study was to compare the pharmacokinetics of acyclovir after intravenous administration and after oral administration of valacyclovir. PROCEDURE: The pharmacokinetics of acyclovir were studied in 18 children aged 1.4-18.1 years (median: 6.9 years; 9 females) after intravenous infusion (1 hr; median dose: 10.5 mg/kg). In 10 of the children the pharmacokinetics of acyclovir were also studied after oral administration of valacyclovir (median dose: 34.1 mg/kg). Quantification of acyclovir in serum was performed by reversed-phase liquid chromatography with fluorometric detection. The pharmacokinetic analysis was performed by pharmacokinetic modelling. RESULTS: The serum concentration versus time curves of acyclovir were described by the two compartment model after intravenous administration and by the one compartment model with a zero- or first-order absorption phase after oral administration of valacyclovir. The bioavailability of acyclovir after oral administration of valacyclovir was 45% (median value; 95% CI: 37-55%). CONCLUSION: It is possible to substitute intravenous acyclovir therapy by oral valacyclovir therapy in children with leukopenia and mucositis after chemotherapy. This finding can at present not be fully implemented in clinical practice, since a commercial pharmaceutical formulation of valacyclovir aimed for children not able to swallow intact tablets is lacking. Crushed valacyclovir tablets have a very unpleasant taste, but can be administered to children through nasogastric tubes. PMID- 11920786 TI - Study design and cohort characteristics of the Childhood Cancer Survivor Study: a multi-institutional collaborative project. AB - BACKGROUND: Increased attention has been directed toward the long-term health outcomes of survivors of childhood cancer. To facilitate such research, a multi institutional consortium established the Childhood Cancer Survivor Study (CCSS), a large, diverse, and well-characterized cohort of 5-year survivors of childhood and adolescent cancer. PROCEDURE: Eligibility for the CCSS cohort included a selected group of cancer diagnoses prior to age 21 years between 1970-1986 and survival for at least 5 years. RESULTS: A total of 20,276 eligible subjects were identified from the 25 contributing institutions, of whom 15% are considered lost to follow-up. Currently, 14,054 subjects (69.3% of the eligible cohort) have participated by completing a 24-page baseline questionnaire. The distribution of first diagnoses includes leukemia (33%), lymphoma (21%), neuroblastoma (7%), CNS tumor (13%), bone tumor (8%), kidney tumor (9%), and soft-tissue sarcoma (9%). Abstraction of medical records for chemotherapy, radiation therapy, and surgical procedures has been successfully completed for 98% of study participants. Overall, 78% received radiotherapy and 73% chemotherapy. CONCLUSION: The CCSS represents the largest and most extensively characterized cohort of childhood and adolescent cancer survivors in North America. It serves as a resource for addressing important issues such as risk of second malignancies, endocrine and reproductive outcome, cardiopulmonary complications, and psychosocial implications, among this unique and ever-growing population. PMID- 11920788 TI - Analysis of TP53 germline mutations in pediatric tumor patients using DNA microarray-based sequencing technology. AB - BACKGROUND: Whereas in sporadic human malignancies mutations of the TP53 tumor suppressor gene occur in cancers of almost every organ and histologic subtype, patients with an inborn TP53 defect are at high risk to develop, in particular, soft tissue and bone sarcomas, brain tumors, leukaemias, adrenocortical tumors, and breast cancer. To demonstrate the usefulness of microarray technology applied to TP53 sequencing in pediatric tumors, we investigated young patients suffering from tumors typical of the Li-Fraumeni context who were, therefore, candidates for harboring inborn defects in tumor-suppressor genes. PROCEDURE: Six individuals were studied, including typical Li-Fraumeni patients as well as patients without any family history of cancer. DNA samples were independently analyzed for TP53 mutations by GeneChip and standard automated laser fluorescence (ALF) sequencing technology. RESULTS: The tumor and corresponding constitutional DNA samples clearly showed identical mutations which were confirmed by ALF sequencing. All coding exons (exons 2-11) of TP53 were analyzed simultaneously. The entire sequencing procedure and data analysis was carried out within 24 hr. CONCLUSIONS: The GeneChip TP53-sequencing assay may be feasible for routine molecular genetic diagnostics in determining the TP53 status of childhood-tumor patients and in enabling a disease management based on the genetic background of the individual. PMID- 11920789 TI - Predictive value of clinical evaluation in the follow-up of children with a brain tumor. AB - BACKGROUND: During follow-up of children with a brain tumor, traditionally surveillance-imaging studies are done in addition to clinical evaluations. The purpose of this study was to determine the role of clinical evaluations by a multidisciplinary team for the detection of recurrent tumor. PROCEDURE: We retrospectively assessed the predictive value of clinical evaluation, using subsequent neuroimaging as the gold standard. Ninety-eight children with a newly diagnosed primary brain tumor were included in the study. In these patients, 393 imaging studies were done, 75 because of clinical suspicion of recurrence, and 318 for routine surveillance only. RESULTS: In 28 of these 75 imaging studies on clinical indications, a radiologic diagnosis of recurrence was made. Only 9 out of 318 routine surveillance imaging studies resulted in an unexpected diagnosis of recurrence. Thus, the overall positive predictive value of clinical evaluation was 37%; the overall negative predictive value was 97%. The negative predictive values for specific brain tumors varied from 91% (optical glioma) to 99% (primitive neuroectodermal tumors). CONCLUSIONS: An accurate multidisciplinary clinical evaluation has a very high negative predictive value. Consequently, the added value of surveillance imaging studies is limited. The role of such studies during the follow-up of children with brain tumors should be reconsidered. PMID- 11920790 TI - Excess risk for Mullerian duct anomalies in girls with Wilms tumor. PMID- 11920791 TI - Verdi and death. PMID- 11920792 TI - Psychological support in children and adolescents with cancer when amputation is required. PMID- 11920793 TI - The treatment of children with acute lymphoblastic leukemia in Thailand. PMID- 11920794 TI - Automated differential counts in acute promyelocytic leukemia patients may be misleading. PMID- 11920795 TI - Predilection of retinoblastoma metastases for the mandible. PMID- 11920796 TI - Spontaneous resolution of isolated thymic Langerhans cell histiocytosis. PMID- 11920797 TI - Retinoblastoma with central retinal artery thrombosis that mimics extraocular disease. PMID- 11920798 TI - Rectal cancer twenty-one years after treatment of childhood Hodgkin disease. PMID- 11920799 TI - Leukemic pericardial effusion causing cardiac tamponade. PMID- 11920800 TI - Leydig cell tumor and mature teratoma: unusual non-ocular tumors associated with sexual pseudo-precocity six years after unilateral retinoblastoma. PMID- 11920801 TI - Pre-eruptive varicella zoster virus encephalitis in two children after haematopoietic stem cell transplantation. PMID- 11920802 TI - Thyroid carcinoma in a fourteen-year-old boy with Graves disease. PMID- 11920803 TI - Assessment and treatment of low-risk febrile neutropenic (LRFN) pediatric patients with cancer. PMID- 11920804 TI - 13q deletion syndrome associated with retinoblastoma and clinical anophthalmos of the opposite eye. PMID- 11920805 TI - High-dose oral ganciclovir for management of CMV-symptomatic infection in a child with acute lymphoblastic leukemia. PMID- 11920806 TI - Meningeal carcinomatosis in a child with Ewing sarcoma. PMID- 11920807 TI - Rituximab with interleukin-2 after autologous bone marrow transplantation for acute lymphocytic leukemia in second remission. PMID- 11920808 TI - Embryonal rhabdomyosarcoma associated with tuberous sclerosis. PMID- 11920809 TI - Infinite follow-up? PMID- 11920811 TI - Cellular tropisms and co-receptor usage of HIV-1 isolates from vertically infected children with neurological abnormalities and rapid disease progression. AB - The longitudinal evolution of HIV-1 phenotypes was studied in a cohort of six vertically infected children with early onset and rapid progression of clinical disease. Among 30 viral isolates obtained from peripheral blood, tropisms for both human blood-derived cells (macrophages, T-lymphocytes), and for human neural (brain-derived) cells (microglia, astrocytes) were determined, as was chemokine co-receptor usage. All children harbored from birth macrophage-tropic isolates using the CCR5 co-receptor. Two children later developed T-cell tropic isolates with CXCR4 and CCR3 usage. While all six patients developed neurological abnormalities, only three produced neural cell tropic isolates, which used CCR5. However, early and persistent finding of both astrocyte- and microglia-tropic isolates in one patient did associate with the most rapid progression to brain atrophy among the six patients. Viral phenotypic properties determined in cell culture did not specifically predict clinical features or course, and the development of AIDS did not coincide with, or depend on, the appearance T-tropic, syncytia-inducing viruses. PMID- 11920812 TI - Concordance between semen-derived HIV-1 proviral DNA and viral RNA hypervariable region 3 (V3) envelope sequences in cases where semen populations are distinct from those present in blood. AB - Sequence analysis of the third hypervariable region (V3) of the envelope gene of the HIV-1 was carried out on HIV proviral and viral populations present in blood and semen. Phylogenetically distinct populations of virus were observed in three of the 10 patients analysed. Although the majority of the viruses were predicted to have an R5 phenotype, amino acid differences between blood and semen-derived virus and provirus sequences were observed at sites previously shown to affect cell tropism. Importantly, the semen proviral population was representative of that observed for cell-free virus. This indicates that seminal fluid mononuclear cells are possible sources for the cell-free virus in found in semen. PMID- 11920813 TI - Risk factors and mechanism of transplacental transmission of hepatitis B virus: a case-control study. AB - Intrauterine hepatitis B virus (HBV) infection has been suggested to be caused by transplacental transmission that cannot be blocked by hepatitis B vaccine. This would decrease the effectiveness of hepatitis B vaccine. This study examined the risk factors and mechanism of transplacental HBV transmission. A case-control study included 402 newborn infants from 402 HBsAg-positive pregnant women. Among these, 15 newborn infants infected with HBV by intrauterine transmission were selected as cases, and the rest as controls. A pathology study included 101 full term placentas from the HBsAg-positive pregnant women above and 14 from HBsAg negative pregnant women. Immunohistochemistry staining and HBV DNA in situ hybridization were used to estimate the association of intrauterine HBV infection and HBV infection in the placentas. HBeAg positivity in mothers' sera (OR = 17.07, 95%CI 3.39-86.01) and threatened preterm labor (OR = 5.44, 95%CI 1.15 25.67) were found to be associated with transplacental HBV transmission. The intrauterine infection rate increased linearly and significantly with maternal serum HBsAg titers (trend test P = 0.0117) and HBV DNA concentration (trend test P < 0.01). Results of the pathology study showed that HBV infection rates decreased gradually from the maternal side to the fetal side (trend test P = 0.0009) in the placental cell layers. There was a significant association between intrauterine HBV transmission and HBV infection in villous capillary endothelial cells (VCEC) in the placenta (OR = 18.46, P = 0.0002). The main risk factors for intrauterine HBV infection are maternal serum HBeAg positivity, history of threatened preterm labor, and HBV in the placenta especially the villous capillary endothelial cells. Previous reports of transplacental leakage of maternal blood causing intrauterine infection are confirmed. In addition, there appears to be a "cellular transfer" of HBV from cell to cell in the placenta causing intrauterine infection. This latter hypothesis needs to be confirmed. PMID- 11920814 TI - Chronic hepatitis C infection: influence of the viral load, genotypes, and GBV C/HGV coinfection on the severity of the disease in a Brazilian population. AB - The distributions of the different genotypes of the hepatitis C virus (HCV) and GBV-C virus (GBV-C/HGV) vary geographically and information worldwide is still incomplete. In particular, there are few data on the distribution of genotypes (and their relationship to the severity of liver disease) in South America. Findings are described in 114 consecutive patients from Northeast Brazil (median age 52 years, range 18-72 years) who had abnormal levels of serum aminotransferases and seropositivity for HCV RNA. The patients were recruited from an outpatient clinic between November 1997 and April 1998. Quantitative HCV RNA and GBV-C/HGV RNA estimations were carried out by double-nested polymerase chain reaction (PCR) using primers from the 5'-untranslated regions (UTRs) of the genomes. HCV genotypes were determined by restriction fragment length polymorphism (RFLP) analysis with 5'-UTR primers and by PCR with type-specific 5' UTR primers. GBV-C/HGV-RNA genotypes were determined by RFLP with specific 5'-UTR primers and phylogenetic trees were constructed using the Neighbour-Joining and Drawtree programs. Histological features were graded and staged according to international criteria. Of the 114 patients, 35 (30.7%) patients had cirrhosis and 22 (27.8%) had mild, 51 (64.6%) had moderate, and 6 (7.6%) had severe chronic hepatitis. Median HCV viral load was 10(6) genome equivalents per millilitre (range 10(4)-10(9)/ml). Frequencies of genotypes were 5.3% type 1a, 44.7% type 1b, 3.5% type 2, 41.2% type 3, and 5.3% mixed types. GBV-C/HGV-RNA was detected in the sera of 12 (10.5%) patients and was distributed among three phylogenetic groups. There were no significant differences between patients with the predominant HCV genotypes (1b and 3) with respect to gender, age group, viral load, severity of liver disease, or coinfection with GBV-C/HGV. PMID- 11920815 TI - Human cytomegalovirus hyperimmune globulin not only neutralizes HCMV infectivity, but also inhibits HCMV-induced intracellular NF-kappaB, Sp1, and PI3-K signaling pathways. AB - Inhibition of virus-induced intracellular signaling pathways and viral infectivity are our ultimate goals in the development of effective antiviral agents to control human cytomegalovirus (HCMV) infections. The HCMV hyperimmune globulin may meet such criteria. In a human embryonic lung (HEL) fibroblast culture model, pretreatment of Towne strain HCMV with HCMV hyperimmune globulin was shown to inhibit viral infectivity successfully, as measured by a standard plaque assay. The extracellular viral titers and extracellular viral DNA, as measured by plaque assay and PCR, respectively, were also decreased. In addition, the HCMV hyperimmune globulin prevented HCMV from inducing the intracellular activation of NF-kappaB, Sp-1, and PI3-K signaling pathways. The PI3-K pathway was examined by following phosphorylation (activation) of two of its downstream kinases, Akt and p70S6K. HCMV hyperimmune globulin also prevented the production of immediate early, early, and late viral proteins. These studies show that HCMV hyperimmune globulin neutralization of HCMV prevents the earliest known events observed after viral envelope glycoproteins bind their cell membrane receptors, i.e., NF-kappaB, Sp-1 and PI3-K activation. This suggests that HCMV hyperimmune globulin not only can inhibit viral infectivity, but can also prevent the abnormal cellular signaling that may induce unwanted cellular proliferation or cytokine synthesis. PMID- 11920816 TI - Cytokine gene expression and protein production in peripheral blood mononuclear cells of children with acute dengue virus infections. AB - Plasma leakage in dengue hemorrhagic fever (DHF) is associated with elevated plasma levels of cytokines. To define further the contribution of immune activation to DHF and the source of cytokines, we analyzed the production of cytokines in peripheral blood mononuclear cells (PBMC) obtained from children with dengue, using RT-PCR and immunostaining. Tumor necrosis factor-alpha (TNF alpha) and TNF-beta expression was detected in all samples by PCR and in < 50% of samples by immunostaining. Interferon-gamma (IFN-gamma) expression was detected in < 50% of samples by either method. Interleukin-2 (IL-2) and IL-4 expression was detected in a few samples by immunostaining but was not detectable by PCR. We found greater expression of TNF-alpha and IL-4 in DHF than in dengue fever or other (non-dengue) febrile illnesses. These results support the model of immunopathogenesis of DHF. However, low levels of cytokine expression in PBMC suggest that cellular activation in tissues may contribute to high serum cytokine levels in DHF. PMID- 11920817 TI - Enterovirus meningitis in adults in 1999-2000 and evaluation of clinical management. AB - Enterovirus meningitis is well documented in children. However, there is a paucity of reports in adults, despite the availability of genome detection (RT PCR) in cerebrospinal fluid (CSF), which provides a rapid and reliable diagnosis. The clinical course and management of 30 cases of entero-virus proven meningitis prospectively diagnosed between August 1999 and November 2000 in immunocompetent adults were analysed, and laboratory and clinical strategies evaluated. Patient age ranged between 17 and 43 (median 29). The analysis of clinical, biological, and epidemiological data showed the difficulty of recognising enterovirus meningitis in adults. Characteristic symptoms were either inconstant (the association of fever/headache/stiff neck) or misleading (the presence of vesicular lesions). CSF data showed moderate pleocytosis but a predominance of lymphocytes in only 12/27 (44%) patients. An epidemiological background was present in 10/30 (33%) patients, but 10/30 (33%) patients were admitted during cold months. Consequently, although the detection of enterovirus genome in CSF was positive in all cases, the results were communicated within a median of 6 days [2-9] after admission, mainly because the aetiology was not considered early enough. Management of patients varied between departments and between individual physicians, with measures ranging from computed tomography (33%) to the prescription of aciclovir (20%) or antibiotics (53%). Enterovirus meningitis should not be underestimated in adults. Management could be improved and standardised, and costs reduced by more systematic year-round use of enterovirus RT-PCR in meningitis, provided results are rapid. PMID- 11920818 TI - Epstein-Barr virus (EBV) DNA levels in palatine tonsils and autologous serum from EBV carriers. AB - A real-time polymerase chain reaction was employed to detect and quantitate Epstein-Barr virus (EBV) DNA in tonsils and autologous sera from EBV-seropositive children. EBV DNA was found in 95% of tonsils from 21 children and in 50% from 18 children with serum IgG titers to the virus capsid antigen (VCA) of > or =1:160 and 1:10 to 1:80, respectively (P = 0.002). Tonsils from children with titers > or =1:160 harbored more EBV DNA copies per mg tissue (mean, 1,237; range, < 2 13,998) than from children with titers 1:10 to 1:80 (mean, 23; range, < 2-226; P < 0.0001). By contrast, EBV DNA was detected only in serum from 25% of 20 children with titers > or = 1:160. Thus, ample differences in tonsillar EBV replication are mirrored inconstantly by detectable EBV in autologous serum suggesting that EBV DNA quantitation in tonsils may serve for refined monitoring of individuals at risk of EBV-associated lymphoproliferation. PMID- 11920819 TI - Characteristics of Epstein-Barr virus isolated from the malignant lymphomas in Korea. AB - Epstein-Barr virus (EBV) is a common herpes virus linked to a variety of human neoplasms. In this study, the EBV detection was identified with the paraffin embedded tissues from 62 non-Hodgkin's lymphomas, 20 Hodgkin's lymphomas, and 48 non-neoplastic tonsils, using PCR for EBNA-1 and EBER-1 mRNA in situ hybridization for EBER-1 mRNA. The isolates were analyzed for type 1/2, variants C/D and F/f, and LMP-1 30 bp deletion. EBV was isolated in 31 of 48 (66%) non neoplastic tonsils, 24 of 42 (57%) B cell lymphomas, in 15 of 20 (75%) T cell lymphomas, and 17 of 20 (85%) Hodgkin's lymphomas. These viruses were classified as type 1 for 81% of non-neoplastic tonsils, 95% of B cell lymphomas, 93% of T cell lymphomas, and 73% of Hodgkin's diseases. Both type 1 and 2 viruses were isolated in one non-neoplastic tonsil and 3 Hodgkin's diseases. Type C virus was predominant in non-neoplastic tonsils (77%) and B cell lymphomas (75%), while type D virus was common in T cell lymphomas (71%) and Hodgkin's diseases (73%) (P < 0.05). Majority of the viruses detected in non-neoplastic tonsils (93%) and malignant lymphomas (91%) were "F" prototype. LMP-1 30 bp deletion was found in high frequency in both non-neoplastic tonsils (92%) and malignant lymphomas (86%). In conclusion, most of EBV found in Korea was type 1, and "DF" genotype was more frequent in T cell lymphomas and Hodgkin's diseases than in non neoplastic tonsils and B cell lymphomas. LMP-1 30 bp deletion did not seem to be associated with malignant lymphomas. PMID- 11920820 TI - Human herpesvirus 6 variant a infects human term syncytiotrophoblasts in vitro and induces replication of human immunodeficiency virus type 1 in dually infected cells. AB - Human herpesvirus 6 (HHV-6) and human immunodeficiency virus type 1 (HIV-1) may interact during transplacental transmission of HIV-1. The placental syncytiotrophoblast layer serves as the first line of defense of the fetus against viruses. Patterns of replication of HHV-6 variant A (HHV-6A) and HIV-1 were analyzed in singly and dually infected human term syncytiotrophoblast cells cultured in vitro. For this purpose, the GS strain of HHV-6A and the Ba-L and IIIB strains of HIV-1 were used. HHV-6A replication was restricted at the level of early gene products in singly infected syncytiotrophoblasts, whereas no viral protein expression was found in cells infected with HIV-1 alone. Coinfection of syncytiotrophoblast cells with HHV-6A and HIV-1 resulted in production of infectious HIV-1. In contrast, no enhancement of HHV-6A expression was observed in cell cultures infected with both viruses. Uninfected syncytiotrophoblast cells were found to express CXCR4 and CCR3 but not CD4 or CCR5 receptors. Infection of syncytiotrophoblasts with HHV-6A did not induce CD4 expression and had no influence on chemokine receptor expression. Activation of HIV-1 from latency in coinfected cells was mediated by the immediate-early (IE)-A and IE-B gene products of HHV-6A. Open reading frames U86 and U89 of the IE-A region were able to activate HIV-1 replication in a synergistic manner. The data suggest that in vivo double infection of syncytiotrophoblast cells with HHV-6A and HIV-1 could contribute to the transplacental transmission of HIV-1 but not HHV-6A. PMID- 11920821 TI - Herpes simplex virus isolates with reduced adefovir susceptibility selected in vivo by foscarnet therapy. AB - Sequential herpes simplex virus (HSV) isolates from AIDS patients receiving foscarnet therapy were evaluated for susceptibility to adefovir. Foscarnet resistant isolates with DNA polymerase mutations in regions II, VI, and between I and VII were also associated with an important decrease in susceptibility to adefovir (mean IC50 increase: 4.6-fold compared to pre-foscarnet or wild-type isolates) suggesting that adefovir-resistant HSV could be selected in vivo by foscarnet therapy. PMID- 11920823 TI - Molecular epidemiology of rotaviral infection in South Indian children with acute diarrhea from 1995-1996 to 1998-1999. AB - The distribution of VP7 (G-) and VP4 (P-) genotypes among 126 rotavirus strains from South Indian children, < 5 years of age and with acute diarrhoea, presenting to a single hospital during the months to November and December, from 1995 to 1998, was studied. Multiplex hemi-nested G- and P-typing polymerase chain reactions determined 101 (80%) G types and 78 (61%) P types, respectively. In order of frequency, the commonest G types were G1, G4, G2, G9, G3, and G8, and P types were P1B[4], P1A[8], and P2A[6] and the most common G:P combinations were G1:P1A[8], G1:P1B[4], G2P1B[4] and G4:P1A[8]. G1, G2, and G4 types were seen in all years. The single G3 isolate was seen in 1998. The single G8 isolate and the 5 G9 isolates were seen in 1997, after a period of heavy rain. Sequence analysis showed that the G8 isolate was related most closely to the bovine strain A5, and the G9 strains were distinct from the nonpathogenic Indian isolate 116E and similar to G9s isolated in Mysore and the United Kingdom described previously. PMID- 11920822 TI - Genetic susceptibility to respiratory syncytial virus infection in inbred mice. AB - Differences in the severity of respiratory syncytial virus (RSV)-induced lower respiratory disease in infants have been attributed to multiple environmental and genetic factors. To identify the genetic factor(s) influencing RSV susceptibility, we examined RSV infection in eight inbred mouse strains. Lung RSV titers differed significantly between mouse strains: the RSV titers were 15-fold higher in AKR/J (permissive) mice compared with C57BL/6J (resistant) mice at 4 days after inoculation. This strain-specific difference in RSV titers suggested that susceptibility to RSV infection was attributable to genetic differences between strains. To examine the mode of inheritance of RSV susceptibility, F1 and backcross (F1 x AKR/J) progeny were infected and RSV titers determined. RSV titers in the F1 progeny were similar to those found in the resistant (C57BL/6J) parent, suggesting resistance was inherited as a dominant trait. The distribution of RSV titers in backcross progeny were discordant with that predicted for a single gene effect, suggesting susceptibility was influenced by more than one gene. These data suggest that RSV susceptibility is a multigenic trait that should be amenable to resolution by genomic analysis. PMID- 11920824 TI - Incidence and prevalence of human group C rotavirus infections in Argentina. AB - The incidence of human group C rotavirus infections among children and adults in Buenos Aires was evaluated by enzyme linked immunosorbent assays (ELISA) based on recombinant group C VP6 protein (Cowden strain). A total of 976 stool samples taken from patients (ages 6 months to 15 years) with acute diarrhea were tested for the presence of group C rotavirus. Among these, only 10 (1.02%) were group C rotavirus positive by enzyme-linked immunosorbent assay (ELISA) confirmed by absorption with group C VP6 antibodies and by RT-PCR for both VP6 and VP7 genes. The average age (5.86 years) was significantly superior to that in group A infected patients (1.63 years). Previous exposure to this virus was assessed by detecting specific IgG in sera taken from healthy individuals grouped by age. Of 844 sera tested, 425 (50.3%) were group C IgG positive by ELISA, confirmed by Western blot analysis. The rates of IgG positivity for group A and C rotaviruses during the first years of life indicated that infections with group C are frequent in older children (3-5 years), whereas group A infections are prevalent in infants and young children (6-18 months). This study shows that group C rotavirus infections in Argentine children occur later in life than group A and are relatively common in spite of the low detection rate of this virus. PMID- 11920825 TI - Novel TTV variants isolated in an epidemic of hepatitis of unknown etiology. AB - TT virus (TTV) is a recently discovered single-stranded DNA virus that has been reported to be associated with elevated transminase levels in the patients with posttransfusion hepatitis of unknown etiology. TTV prevalence is very high in the common population and its pathogenicity remains unclear. In this study, we performed an epidemiological study to investigate the infection rate of TTV and its role in an epidemic of unknown-etiology hepatitis. Moreover, two TTV isolates named L01 and L02 were cloned from the serum of a patient with unknown-etiology hepatitis. Eighty-one subjects were included in the study and were divided into two groups: 18 in the case group and 63 in the control group. TTVDNA was detected by nested PCR from sera samples. The infection rates of TTV in case and control groups were 33.3 and 38.9%, respectively. There was no significant difference between the two groups. Homology analysis showed that L01 had a very poor homology with other TTV isolates and L02, and L02 was 75.5% identical to JA10. The result does not support TTV as a causative agent in this epidemic. The genetic divergence between L01 and other TTV isolates beyond genotype, so it represents a new genotype of TTV. PMID- 11920826 TI - Novel variants related to TT virus distributed widely in China. AB - TTV is a DNA virus with high genetic heterogeneity. To investigate the novel isolates of the virus, blood samples were collected from subjects who lived in various parts of China and suffered from hepatitis or were asymptomatic carriers. Nested PCR was carried out to amplify a 3.2-kb fragment using primers deduced from the prototype TTV (TA278). The ten entire 3.2-kb nt sequences were aligned with isolate TA278, SANBAN, TUS01, and SENV retrieved from GenBank, and a phylogenetic tree was constructed by Neighbor-Joining method. The analysis indicated that five novel variants of the present study have not been described before, and all TTV-related isolates could be classified into three groups. The isolate TCHN-A, B and TUS01 were included in a group, and the remaining novel isolates together with SANBAN and TA278 clustered into another group, while SEN virus formed a distinct group. The genetic distances of the five novel variants were 0.5507-0.8476 to TA278, 0.4635-0.7877 to SANBAN, 0.6064-0.7834 to TUS01 and 0.6936-0.8236 to SENV. Of these novel variants, the ORF1 consisted of 426-772 aa and ORF2 of 141-156 aa. The nt identities of ORF1 and ORF2 between those variants and TA278, SANBAN, and TUS01 were 46.1-60.8 and 48.7-63.6%, and those of aa sequences were only 27.1-52.4 and 28.9-45.5%, respectively. The first 65 aa of ORF1 were rich in arginine and most conserved with homology of 56.5-70.0%. There was a hypervariable region from aa 286 to 403 with merely 17.7-27.0% of identity. Despite a low aa identity between TA278 and the variants, they have similar hydrophilicity profiles of ORF1. There were 2-10 N-glycosylation motifs found in these variants. In conclusion, despite the high divergence, sequences of all these isolates shared common genome organisation, ORF structure, hydrophilicity patterns, and some potential motifs with TTV prototype. It is suggested that various TTV and TTV-related isolates belong to a very large and complex family, which remains to be studied. PMID- 11920827 TI - Trafficking mechanism of West Nile (Sarafend) virus structural proteins. AB - Previous studies have shown that West Nile (Sarafend) virus matured by budding at the plasma membrane, which differs from the usual intracellular maturation of other flaviviruses. The present study investigated the trafficking mechanism of the envelope (E) and capsid (C) proteins of West Nile (Sarafend) virus during the replication cycle. The use of time-based double-immunofluorescence labelling coupled with the Triton X-100 extraction procedure revealed that both the E and C proteins were transported from the perinuclear region towards the plasma membrane along the microtubules simultaneously. The strong association of these virus proteins with the microtubules was demonstrated further with Triton X-100 extraction procedure coupled with double immunogold-labelling. Extraction of infected cells with Triton X-100 in high salt also revealed that virus E proteins were associated with the microtubules via protein-protein interaction. The disruption of microtubules with vinblastine sulphate inhibited the trafficking of both the virus E and C proteins. Both virus structural proteins were observed to co-localise and retained within vinblastine sulphate-induced microtubulin paracrystals. Extracellular virus production was also reduced drastically by vinblastine sulphate at non-cytotoxic concentration. Subsequent studies revealed that the transportation of virus E protein was associated with the microtubules based motor protein, kinesin. PMID- 11920828 TI - Vascular birthmarks of infancy: resolving nosologic confusion. AB - The terminology describing congenital vascular birthmarks has been a source of confusion in the medical literature. Mulliken and Glowacki [1982: Plas. Recons. Surg. 69:412-422] published a biologic classification system which has become the most widely accepted framework for classifying vascular birthmarks and is accepted as the official classification schema by the International Society for the Study of Vascular Anomalies (ISSVA). In this study, we evaluate the current nosology of vascular birthmarks used in standard medical genetics reference texts compared with the accepted Mulliken ISSVA framework. In five sources examined, a variety of terms were used to describe congenital vascular anomalies. The degree of agreement with accepted ISSVA classification varied both within and among texts, with agreement as low as 22% and as high as 75%. In all texts, hemangioma was the most commonly used term, appearing 79 times. Use of the term "hemangioma" had the lowest rate of agreement with the ISSVA classification criteria, with agreement in 23% of citations. The terms "vascular malformation" and "port-wine stain" were used less frequently, but with a much higher degree of agreement with the ISSVA classification: 82% and 66%, respectively. These results establish that nosologic confusion is widespread even in standard genetic reference texts. In particular, the term "hemangioma" is used imprecisely. The ISSVA classification system provides an extremely useful framework for geneticists to classify vascular birthmarks in their evaluation of infants and children with vascular anomalies in order to provide more accurate evaluation, prognosis, and genetic counseling. PMID- 11920829 TI - Vasculogenesis, angiogenesis, hemangiomas, and vascular malformations. PMID- 11920830 TI - Van der Woude syndrome with sensorineural hearing loss, large craniofacial sinuses, dental pulp stones, and minor limb anomalies: report of a four generation Thai family. AB - A four-generation Thai family affected with Van der Woude syndrome is reported. The disorder appeared to be originally inherited from a person who was half Thai and half Pakistani. The lip lesions found in this family were varied and did not appear to be related to other phenotypes. There were some clinical manifestations possibly specific for the condition in this family. They included sensorineural hearing loss, prominent frontal bone, large frontal/sphenoidal/maxillary sinuses with increased mastoid air cells, long tooth roots, dental pulp stones, ankyloglossia, brachydactyly of hands, brachyphalangy, and hyperphalangy of toes, and single flexion crease of the fifth fingers. Fluorescence in situ hybridization analysis revealed no visible deletion at a 1q32-41 region. PMID- 11920831 TI - Mixed clefting type in Rapp-Hodgkin syndrome. AB - Mixed clefting type (MCT) is the rare occurrence of cleft lip, with or without cleft palate, and cleft palate alone in the same pedigree. Here we present a family with Rapp-Hodgkin syndrome (RHS) that manifests MCT, and use this rare finding to suggest that RHS may be related not only to phenotypically similar syndromes, but seemingly dissimilar ones as well. RHS has obvious phenotypic overlap with other ectodermal dysplasia-clefting syndromes (EDCS), such as ectrodactyly-ectodermal dysplasia-clefting syndrome (EEC) and ankyloblepharon ectodermal dysplasia-clefting syndrome (AEC), all of which show MCT. MCT is also found in the allelic disorders van der Woude syndrome (VDW) and popliteal pterygium syndrome (PPS). Therefore, while VDW and PPS have little clinical overlap with the EDCS, the common finding of MCT may indicate closer relationships at the developmental or genetic level. PMID- 11920832 TI - Prediction by FISH analysis of the occurrence of Wilms tumor in aniridia patients. AB - Aniridia is an autosomal dominant eye anomaly caused by haploinsufficiency of the PAX6 gene, of which abnormalities include base alterations, position effects and deletions. When deletion involves its adjacent genes, i.e., those in the PAX6-WT1 critical region (WTCR), patients are predisposed to Wilms tumor. We studied 18 patients with aniridia, five of whom had chromosome deletion involving 11p13, two a translocation t(10;11)(p13;p13) or a der(14;21)(q10;q10)mat, and 11 had a normal karyotype. Fluorescence in situ hybridization (FISH) using four P1-derived artificial chromosome (PAC) clones located at WTCR was carried out in the 18 patients to identify a deletion extent. Of the 18 patients, eight had a deletion of WTCR: four had microscopic deletion and four a deletion of WTCR. Deleted region in one patient with a microscopic deletion was distal to the critical region. Four of the eight patients with a deletion encompassing WTCR developed Wilms tumor, and the other four did not (two were too young to be evaluated for the tumor development). The data in the present study, together with four similar previous works, indicate that of a total of 102 aniridia patients, 29 had a deletion spanning WTCR. Wilms tumor developed in 13 (45%) of the 29 patients, whereas patients without deletion in this region did not develop the tumor. In other words, aniridia patients with WT1 deletion run a high risk of developing Wilms tumor, and those without the deletion do not. PMID- 11920833 TI - Short bi-iliac distance in prenatal Ullrich-Turner syndrome. AB - The purpose of the present study is to evaluate the bi-iliac distance and the caudo-cranial position of the iliac bones in Ullrich-Turner syndrome (UTS) fetuses compared to recently published standards for normal fetuses. Whole-body radiographs in antero-posterior projections of 24 UTS fetuses (crown-rump lengths, 106-220 mm) were included in the study. From each radiograph, two horizontal (outer and inner bi-iliac distances) and two vertical (caudo-cranial) positions compared to the vertebral column were measured to estimate the position of the iliac bones. The present investigation revealed that both the outer and inner bi-iliac distances were significantly shorter in UTS fetuses than in normal fetuses. We also found that for the inner bi-iliac distance, the growth rate in UTS fetuses was significantly lower than in normal fetuses. This finding suggests not only a lesser growth but also a different growth pattern compared to normal fetuses. Regarding the caudo-cranial position of the iliac bones compared to the lower vertebral column, there was no significant difference for the lower caudo cranial position, but the upper caudo-cranial position was significantly lower in UTS fetuses than in normal fetuses. The bi-iliac distance and the iliac bone position have not previously been described in Ullrich-Turner syndrome fetuses. PMID- 11920835 TI - Further evidence for a third deafness gene within the DFNA2 locus. AB - DFNA2 is a complex locus. Two hearing loss genes have been identified at this site: GJB3, the gene that encodes the gap junction protein connexin 31, and KCNQ4, a voltage-gated potassium channel gene. A third gene has previously been postulated to explain the hearing loss in an Indonesian family linked to the region but devoid of mutation in either known gene (Van Hauwe et al. [1999: Nat Genet 21:263]). We have identified a large five-generation family with nonsyndromic, autosomal dominant progressive high-frequency hearing loss. The hearing impairment maps to 1p34, the site of the DFNA2 locus. Two-point linkage analysis of microsatellite markers spanning the locus resulted in a lod score of 6.6 at D1S391 at theta = 0. We have investigated both identified deafness genes in affected and unaffected family members and have not found any disease-causing mutations, suggesting that another hearing impairment gene resides at the DFNA2 locus. PMID- 11920834 TI - Anticipation in a unique family with Charcot-Marie-Tooth syndrome and deafness: delineation of the clinical features and review of the literature. AB - Charcot-Marie-Tooth disease (CMT) is a clinically and genetically heterogeneous group of polyneuropathies characterized by degeneration of peripheral nerves, resulting in distal muscle atrophy, sensory loss, and deformities of hands and feet. We have studied 34 individuals in a large 84-member four-generation central Illinois family with autosomal dominant Charcot-Marie-Tooth and deafness. Nerve conduction velocities are consistent with type 1 CMT. Audiological evaluation revealed both auditory neuropathy and cochlear involvement in affected individuals. There is increasing clinical severity and younger age of onset of CMT and deafness with each progressive generation, suggestive of anticipation (P < 0.05). The proband, a female diagnosed at birth with hypotonia, bilateral vocal cord palsy, swallowing incoordination, and hearing impairment, died at age 18 months. Another individual died at the age of 3 months from hypotonia later attributed to CMT. Genetic analysis indicated that affected individuals in this family do not have the common 1.4 Mb duplication associated with type 1A CMT; however, all affected individuals have a unique G to C transversion at position 248 in coding exon 3 of the peripheral myelin PMP22 gene located on chromosome 17p11.2-p12. This mutation is predicted to cause an Ala67Pro substitution in the second transmembrane domain of PMP22, consistent with the molecular cause of the CMT phenotype. However, it does not explain the cochlear component of the deafness, the clinical observation of anticipation, and other features in this family. PMID- 11920836 TI - Glypican 1 gene: good candidate for brachydactyly type E. AB - In this report, we describe two siblings, a brother and a sister, with mental retardation and limb abnormalities (brachymetacarpy and brachymetatarsy in the brother and clinodactyly in his sister). Fluorescent in situ hybridization analysis (FISH) using subtelomeric probes proved that the patients carried an unbalanced subtelomeric rearrangement with 2qter deletion involving the Glypican 1 gene. Given the expression pattern in the developing limb bud, Glypican 1 gene represents a good candidate for brachydactyly E. PMID- 11920837 TI - Kallmann syndrome in a patient with congenital spherocytosis and an interstitial 8p11.2 deletion. AB - We describe the hitherto smallest interstitial 8p11.2 deletion in a patient with congenital spherocytosis, dysmorphic features, and growth delay in association with hypogonadotropic hypogonadism and anosmia. The latter features are characteristic for Kallmann syndrome. In contrast to the previously reported patients with 8p deletions, the present patient showed normal intelligence. Congenital spherocytosis is one of the most common hereditary hemolytic anemias. One of the three loci for congenital spherocytosis was assigned to chromosome 8p (located between 8p11.1 and 8p21) and mutations in or loss of the ankyrin-1 gene (ANK1) were identified. Molecular analysis confirmed the de novo loss of ANK1 in our patient. Kallmann syndrome, which is characterized by hypogonadotropic hypogonadism and anosmia, can be X-linked, autosomal dominant, or autosomal recessive. So far only the X-linked KAL1 gene has been identified. The present finding suggests an autosomal locus for Kallmann syndrome at 8p11.2. The simultaneous occurrence of congenital spherocytosis, Kallmann syndrome phenotype, dysmorphic features, and growth delay in this patient points to a new contiguous gene syndrome. PMID- 11920838 TI - Molecular characterization of 22q11 deletion in a three-generation family with maternal transmission. AB - Haploinsufficiency of chromosome 22q11.2 is a well-established cause of both the DiGeorge anomaly and the velocardiofacial syndrome. This condition shows a continuous spectrum of phenotypic manifestations with a considerable inter- and intrafamilial variability. We report on a three-generation family with four members sharing the same 3 Mb long deletion but showing different phenotypic expression. In the first generation, the deleted patient has hypernasal speech and suffers from recurrent psychotic episodes. Two of her offspring inherited the deletion. One of these, a male, has hypernasal speech, low-set ears, hypocalcemia, severe development delay, and tetralogy of Fallot. The other, a female, has hypernasal speech, minor facial anomalies, and very mild mental retardation. Her daughter has tetralogy of Fallot, velopharyngeal insufficiency, and mild facial anomalies. This family is an example of the widely variable phenotypic expressivity of the 22q11.2 deletion. There is no correlation between the size of the deletion and the phenotypic manifestations. Genetic background and/or environmental factors could explain the different phenotypes observed in the affected members of the family. PMID- 11920839 TI - Girl with combined cellular immunodeficiency, pancytopenia, malformations, deletion 11q23.3 --> qter, and trisomy 8q24.3 --> qter. AB - We describe here a 3-year-old girl demonstrating combined cellular immunodeficiency of B- and T-cells, pancytopenia, multiple anomalies, and severe mental retardation. Cytogenetic analysis and fluorescent in situ hybridization (FISH) indicated an unbalanced translocation of chromosomes 8q and 11q, resulting in monosomy 11q23.3-qter and trisomy 8q24.3-qter. The association of cellular immunodeficiency and partial deletion 11q and/or partial trisomy 8q has not been described previously; however, the 11q deletion has been reported with humoral immunodeficiency or pancytopenia. Some one-third to one-half of patients with partial monosomy 11q were reported to have pancytopenia, which has been related to the absence of the 11q23-q24 region. Our case narrows down the critical interval for thrombo- or pancytopenia to 11q23.3-q24 and excludes both the ATM (which resides on 11q23.1) and the MLL genes as possible candidate genes. We are proposing that haploinsufficiency of the NFRKB gene on 11q24-q25 and/or the ETS-1 proto-oncogene on 11q24 may have caused or contributed to the immunodeficiency (decreased levels of B- and T-lymphocytes) in our patient. PMID- 11920840 TI - Scalp defects, polythelia, microcephaly, and developmental delay: a new syndrome with apparent autosomal dominant inheritance. AB - We report a family with apparent autosomal dominant inheritance of scalp defects, polythelia, microcephaly, and developmental delay. A review of the literature revealed no previous report of this combination of anomalies. We conclude that these patients have a new autosomal dominant syndrome. PMID- 11920842 TI - Mosaic trisomy 8 as a cause of velopharyngeal insufficiency. PMID- 11920841 TI - Novel Igalpha (CD79a) gene mutation in a Turkish patient with B cell-deficient agammaglobulinemia. AB - Mutations that impair early B cell development result in profound antibody deficiency, which is characterized by a paucity of mature B cells and the early onset of recurrent pyogenic infections. Among these inherited early B cell defects, X-linked agammaglobulinemia (XLA) with mutations in Bruton's tyrosine kinase (BTK) gene is mostly identified. Recent studies have shown that mutations in the gene for mu heavy chain (IGHM) and for other components of the pre-B cell receptor complex, including lambda5/14.1 (IGLL1) or Igalpha (CD79a), can cause a disorder that is clinically similar to XLA. In a genetic survey of XLA in Turkey, we examined possible mutations in the IGHM, IGLL1, and Igalpha genes in some male patients with presumed XLA who did not have identifiable BTK mutations. We found an eight-year-old boy with a novel homozygous mutation in the Igalpha gene (IVS2+1G>A) causing B cell defect. This is the second case of agammaglobulinemia due to an Igalpha (CD79a) deficiency in the world. PMID- 11920843 TI - Two-loci ADA haplotypes in autistic disorder. PMID- 11920844 TI - The norepinephrine transporter gene and attention-deficit hyperactivity disorder. AB - The adrenergic system plays a known role in attentional systems and a suspected causal role in attention-deficit hyperactivity disorder (ADHD), based on evidence from pharmacological interventions and animal models. The efficacy of the highly selective noradrenergic reuptake inhibitor, tomoxetine, in treating ADHD symptoms supports the system's role in ADHD and points to the norephinephrine transporter as a candidate gene. This study tested the gene for the norepinephrine transporter (NET1) as a susceptibility factor in ADHD using three polymorphisms located in exon 9, intron 9, and intron 13. We examined the inheritance of these polymorphisms in a sample of 122 families with a total of 155 children with ADHD identified through an ADHD proband. Use of the transmission disequilibrium test failed to show significant evidence for biased transmission of any of the alleles or the haplotypes of these polymorphisms. We further investigated this gene by screening the probands for five known amino acid variants to determine if they contributed to the ADHD phenotype but observed only one (Thr99Ile) in our sample. Since the frequency of this variant (1.8%) was similar to that previously reported in a control sample (2.2%), it is unlikely that this variant is related to the ADHD phenotype. Our results do not support the NET1 gene as a major genetic susceptibility factor in ADHD. PMID- 11920845 TI - Quantitative trait locus for reading disability on chromosome 6p is pleiotropic for attention-deficit/hyperactivity disorder. AB - Comorbidity is pervasive among both adult and child psychiatric disorders; however, the etiological mechanisms underlying the majority of comorbidities are unknown. This study used genetic linkage analysis to assess the etiology of comorbidity between reading disability (RD) and attention-deficit hyperactivity disorder (ADHD), two common childhood disorders that frequently co-occur. Sibling pairs (N = 85) were ascertained initially because at least one individual in each pair exhibited a history of reading difficulties. Univariate linkage analyses in sibling pairs selected for ADHD from within this RD-ascertained sample suggested that a quantitative trait locus (QTL) on chromosome 6p is a susceptibility locus for ADHD. Because this QTL is in the same region as a well-replicated QTL for reading disability, subsequent bivariate analyses were conducted to test if this QTL contributed to comorbidity between the two disorders. Analyses of data from sib pairs selected for reading deficits revealed suggestive bivariate linkage for ADHD and three measures of reading difficulty, indicating that comorbidity between RD and ADHD may be due at least in part to pleiotropic effects of a QTL on chromosome 6p. PMID- 11920846 TI - Association study of a SNAP-25 microsatellite and attention deficit hyperactivity disorder. AB - Several lines of evidence implicate synaptosomal-associated protein of 25 kDa (SNAP-25) in the etiology of attention deficit hyperactivity disorder (ADHD). Most notably, the coloboma mouse mutant, considered to be a good animal model of hyperactivity, has a deletion spanning this gene. Introducing a SNAP-25 transgene into these animals alleviates hyperlocomotion. We have identified a novel microsatellite repeat in SNAP-25 located between the 5'UTR and the first coding exon, and tested for association with ADHD. Case-control analyses suggest there may be a role of this polymorphism in ADHD, with one allele over-represented in controls and another over-represented in probands. Within-family tests of linkage and association confirmed these findings. Further work is needed to ascertain the role of SNAP-25 in ADHD and assess the functional significance of this polymorphism. PMID- 11920847 TI - Report from the third international meeting of the Attention-Deficit Hyperactivity Disorder Molecular Genetics Network. AB - Growing evidence of a genetic contribution to attention-deficit hyperactivity disorder (ADHD) has prompted investigators from around the world to convene annually to discuss ways of facilitating collaboration and sharing information about their work on this topic. The number of participants in the meeting has grown each year as a result of enthusiastic responses to each previous conference. This third annual meeting, held in Boston, began with presentations of ongoing and proposed collaborations. The status of Hypescheme, an operational criteria checklist developed in an effort to promote the reliable communication of information related to ADHD, was reviewed. A symposium was conducted to review current evidence for whether DSM-IV subtypes breed true. Finally, new data from individual research sites were presented. Despite recent advances, more work is needed to better characterize heritable aspects of the ADHD phenotype as well as the actual candidate genes themselves. PMID- 11920848 TI - Transmission disequilibrium studies of the serotonin 5-HT2A receptor gene (HTR2A) in autism. AB - Hyperserotonemia in autism is one of the longest-standing biochemical findings in a psychiatric disorder. This well-replicated finding and subsequent studies of platelet serotonin receptors in autism indicate that the serotonin 2A receptor gene (HTR2A) on chromosome 13q is a primary candidate gene in autism. Converging data from recent genome screens also implicates the genomic region containing HTR2A. Based on these lines of evidence, the transmission/disequilibrium test (TDT) was used to assess transmission disequilibrium between autism and haplotypes of three polymorphisms, including the promoter -1438 G/A single nucleotide polymorphism (SNP) in perfect linkage disequilibrium with the 102 T/C SNP in previous studies, a newly identified SNP in intron 1 near exon 2, and the SNP responsible for the His452Tyr amino acid change in exon 3. Because expression studies have shown HTR2A to be polymorphically imprinted in the brain, secondary analyses were split into maternal and paternal transmissions. No evidence was found for unequal transmission of haplotypes; however, power analysis reveals low power to detect a parent-of-origin effect in this sample size. PMID- 11920849 TI - Family-based and population study of a functional promoter-region monoamine oxidase A polymorphism in autism: possible association with IQ. AB - Although the etiology of autism remains to be elucidated, genetic elements significantly contribute to this disorder, and genes on the X chromosome are of special interest because there is a 4:1 predominance of male probands in autism. In the current study, we therefore examined, using the robust transmission disequilibrium test (TDT), possible preferential transmission of variants of a functional monoamine oxidase A (MAO A) promoter region polymorphism for linkage to autism. In the 49 families examined (33 families with one proband and 15 families with two affected siblings), we did not find preferential transmission of MAO A from 33 heterozygous mothers to affected child (TDT chi-square = 0.29, NS). Nor was any significant difference in MAO A allele frequency observed between 43 male autism subjects versus a group of 108 non-autism control subjects (chi-square = 1.23, P = 0.27, NS). However, a trend was observed for an association between IQ in the probands and the MAO A genotype that just attained significance (F = 3.5, P = 0.046, N = 28) in the small group of autism subjects recruited from families with two affected siblings. PMID- 11920850 TI - ApoE epsilon3-haplotype modulates Alzheimer beta-amyloid deposition in the brain. AB - ApoE epsilon4 allele increases the risk of late-onset Alzheimer disease (AD) as well as the amount of beta-amyloid deposition in the brain. Because half of AD patients do not have ApoE epsilon4, it is important to search for other determinants of ApoE that modify AD risk. We tested whether the haplotype background of the most common ApoE allele, epsilon3, influences brain amyloid deposition or the risk of neuropathologically verified AD in a population-based sample of elderly Finns. To exclude the effects of ApoE protein polymorphism we focused these analyses on subjects homozygous for epsilon3. Haplotypes were defined using polymorphisms at positions - 491 and -219 of the ApoE promoter and at position +113 of intron-1. We found that epsilon3-haplotypes containing the promoter allele -219T were associated with reduced amyloid deposition and reduced risk of neuropathologically verified AD as compared to epsilon3-haplotypes containing -219G. The functional polymorphism(s) responsible for the haplotypic difference remains to be identified. These results indicate that there is significant allelic variation in the ApoE gene region, which modulates brain amyloid deposition and AD risk, independent of the ApoE protein polymorphism. PMID- 11920851 TI - Molecular evidence of presenilin 1 mutation in familial early onset dementia. AB - Early onset familial Alzheimer disease (FAD) has been associated with mutations in three genes, of which presenilin 1 (PSEN1) mutations are the most frequent. We reported previously a variant form of FAD, due to deletion of exon 9 of PSEN1, with spastic paralysis and rigidity. We describe a novel PSEN1 mutation in a family of Japanese origin with six affected individuals of both genders in two generations. The disease is characterized by presenile dementia, which is preceded by spastic paraparesis and apraxia. This mutation, which is predicted to cause a missense substitution of serine for glycine, occurred at codon 266 in exon 8 of PSEN1. The mutation was not found in 200 controls and 200 sporadic AD patients. On this basis alone, it seems this mutation is pathogenic. Our findings provide a new clue to the etiology of the familial early onset dementia. PMID- 11920852 TI - Paternal age and sporadic schizophrenia: evidence for de novo mutations. AB - Schizophrenia is an etiologically heterogeneous syndrome. It has a strong genetic component and exists in clinically indistinguishable familial and nonfamilial (sporadic) forms. A significant role for de novo genetic mutations in genetic schizophrenia vulnerability is suggested by a strong monotonic increase in schizophrenia risk with advancing paternal age. However, an alternative explanation for the paternal age effect in schizophrenia is that childbearing is delayed in fathers who themselves have genetic schizophrenia vulnerability. In this study, we compared paternal birth ages between patient groups with familial (n = 35) and sporadic (n = 68) patients with DSM-IV schizophrenia from an inpatient schizophrenia research unit. If later age of fathering children is related to having some genetic schizophrenia vulnerability, then paternal birth age should be later in familial schizophrenia cases than in sporadic cases, and any association of father's age and schizophrenia risk in offspring would be a spurious finding, unrelated to etiology. However, if de novo mutations cause sporadic schizophrenia, then patients without a family history of schizophrenia would have older fathers than familial patients. We found that patients without a family history of schizophrenia had significantly older fathers (4.7 years) than familial patients; so later childbirth was not attributable to parental psychiatric illness. These findings support the hypothesis that de novo mutations contribute to the risk for sporadic schizophrenia. PMID- 11920853 TI - Novel polymorphisms in the promoter region of the neurotrophin-3 gene and their associations with schizophrenia. AB - Based on the neurodevelopmental hypothesis in the etiology of schizophrenia, neurotrophic factors may be involved in the pathogenesis of the illness. We searched for polymorphisms in the promoter region of the neurotrophin-3 (NTF3) gene by using denaturing high performance liquid chromatography (DHPLC). Six single nucleotide polymorphisms (SNPs) were found. When these polymorphisms were examined for association with schizophrenia, a weakly significant difference was observed in the genotype distribution of the G/- 3004/A polymorphism between 184 schizophrenics and 185 controls (P < 0.05), although no statistically significant association was detected in a family-based sample of 50 trios (schizophrenics and their parents). With respect to the other polymorphisms, there was no significant association with schizophrenia. The G/- 3004/A polymorphism was in linkage disequilibrium with the CA repeat polymorphism in the first intron of the NTF3 gene. When haplotype-based analysis was performed, an increased frequency of the haplotype containing the G(- 3004) and the "A3" ([CA]23) alleles was observed for the schizophrenics compared to controls. Our results suggest that the G(- 3004) A3 haplotype has a modest effect of giving susceptibility to schizophrenia. PMID- 11920854 TI - Angiotensin converting enzyme gene insertion/deletion polymorphism: case-control association studies in schizophrenia, major affective disorder, and tardive dyskinesia and a family-based association study in schizophrenia. AB - Angiotensin converting enzyme (ACE) is a candidate gene for psychiatric disorders. We examined the frequency of a functional insertion/deletion (I/D) polymorphism in the 16th intron of the ACE gene (located on chromosome 17q23) in groups of patients with schizophrenia (n = 104 and 113), major depression (n = 55), and bipolar disorder (n = 87) compared to healthy control subjects (n = 87). There was no evidence for allelic or genotypic association of the polymorphism with any of the disorders or with tardive dyskinesia (TD) in patients with schizophrenia. In a sample of nuclear families (n = 61) made up of one or more patients with schizophrenia recruited with their parents, there was no evidence for biased transmission of ACE I/D alleles. Particularly in the case of schizophrenia, these findings do not support an association of the ACE I/D polymorphism with the phenotypes examined. PMID- 11920856 TI - No evidence for association of the T102C polymorphism in the serotonin type 2A receptor with suicidal behavior in schizophrenia. PMID- 11920855 TI - Investigation of linkage and association/linkage disequilibrium of HLA A-, DQA1-, DQB1-, and DRB1-alleles in 69 sib-pair- and 89 trio-families with schizophrenia. AB - The hypothesis that HLA antigens confer susceptibility to schizophrenic disorders has been tested by studying linkage and association in a family sample with 69 sib-pair families. Suggestive evidence for linkage was obtained by nonparametric multipoint LOD score analysis with a maximum around DQB CAR (P = 0.0004), a microsatellite marker that is in linkage disequilibrium with the HLA antigen DQB1. Spurious evidence for negative association as calculated by the transmission disequilibrium test was found for HLA- DRB1*11 (chi-square = 11.72, corrected P value = 0.03) and for the haplotype DQB1*301-DQA1*501-DRB1*11 (chi square = 11.3, corrected P value = 0.043). No evidence of association with these alleles was obtained in a sample of 89 trios with schizophrenic offspring and parents. Our results are not in favor of a direct involvement of the HLA system in development of schizophrenia, but are compatible with the possible existence of a susceptibility gene in the MHC region at chromosome 6p 21.31. PMID- 11920857 TI - Correlation between serotonin uptake in human blood platelets with the 44-bp polymorphism and the 17-bp variable number of tandem repeat of the serotonin transporter. AB - Dysfunctions of the central serotonin (5-hydroxytryptamine, 5-HT) system seem to be associated with psychiatric disorders such as schizophrenia or depression. Previous studies suggested that a 44-bp insertion/deletion polymorphism of the 5 HT transporter (5-HTT) promoter region might influence the transcriptional activity of the 5-HTT gene, and the insertion variant resulted in increased 5-HTT expression and 5-HT uptake. Moreover, a 17-bp variable number of tandem repeat (VNTR) polymorphism of the second intron may act as a transcriptional regulator with allele dependent differential enhancer-like properties. Since the 5-HTT of human platelets shares many properties with the transporter of neural tissue, platelets are widely used as a surrogate tissue source, possibly reflecting central 5-HT metabolism. Therefore, we investigated the impact of the 44-bp polymorphism and the 17-bp VNTR for 5-HT uptake in platelets of 50 male subjects. We found no significant effect of the 44-bp polymorphism and of the 17-bp VNTR on maximum rate (Vmax) of 5-HT uptake. However, individuals homozygous for the 5-HTT intron 2 allele with 12 repeats (STin2.12) of the 17-bp VNTR appeared to have lower affinity of 5-HT uptake than individuals heterozygous for the STin2.10/STin2.9 allele. This was also observed for the combined analysis of both polymorphisms. In conclusion, we found no association between the different genotypes of the 44-bp polymorphism and the 17-bp VNTR and maximum rate of 5-HT uptake into platelets. PMID- 11920858 TI - Allelic association analysis of the dopamine D2, D3, 5-HT2A, and GABA(A)gamma2 receptors and serotonin transporter genes with heroin abuse in Chinese subjects. AB - Five candidate genes, the receptors DRD2, DRD3, HTR2A and GABA(A)gamma2, and the serotonin transporter (5-HTT) were analyzed for association with heroin abuse. We examined three polymorphisms (promoter - 141DeltaC, Ser311Cys, and TaqI) in the DRD2 gene, one polymorphism (Ser9Gly) in the DRD3 gene, two polymorphisms (promoter - 1438G/A and T102C) in the HTR2A gene, two polymorphisms (VNTR and Del/Ins) in 5-HTT gene, and one polymorphism (G3145A) in GABA(A)gamma2 gene in 121 Chinese heroin addicts and 194 controls. None of the polymorphisms differed significantly for allele, genotype, or haplotype frequencies, except for the DRD2 promoter polymorphism - 141DeltaC (genotype-wise and allele-wise, P = 0.05, uncorrected). An additional 344 subjects with heroin abuse and 104 controls were investigated for the - 141DeltaC polymorphism. In the second sample, there were no significant difference of genotype or allele frequencies between subjects with heroin abuse and normal controls. When we divided the sample by route of administration into nasal inhalers and IM or IV injectors, however, it produced a significant difference between inhalers of heroin and controls (genotype-wise, P = 0.006, allele-wise, P = 0.016) but not for injectors of heroin (genotype-wise, P = 0.81, allele-wise, P = 0.69). We also found that LD between all polymorphisms we examined in the gene was weak, possibly explaining why we see association of this polymorphism with heroin abuse but not with other markers in the gene. Overall our results indicates that the HTR2A, 5-HTT, DRD3 and GABA(A)gamma2 genes are not likely to be a major genetic risk factor for heroin abuse in this population, with the exception of possible association between nasal inhalation and DRD2 promoter - 141DeltaC polymorphism. PMID- 11920859 TI - Serotonin 2a receptor T102C polymorphism and impaired impulse control. AB - Patients homozygous for the C allele of the T102C serotonin (5-HT) 2a receptor polymorphism have shown increased suicidal ideation or behavior in some reports, but not in others. We conducted a pilot investigation to determine whether this polymorphism might relate more specifically to a dimension of impaired impulse control, which may underlie only a portion of suicides. Rates of commission (impulsive) errors in a variant of the Continuous Performance Test (CPT) were compared across the genotypes of the T102C polymorphism in adults recruited from the community. The 102C/102C genotype was jointly associated with a greater incidence of past mood disorder or substance-use disorder, as well as significantly more commission errors compared to the 102T/102C and 102C/102C genotypes. These preliminary data suggest that the T102C 5-HT2a receptor polymorphism may be a marker for impaired behavior control-perhaps in the context of psychiatric disorder history. PMID- 11920860 TI - No evidence of an association between a functional monoamine oxidase a gene polymorphism and completed suicides. AB - Monoamine oxidase A (MAOA) has been implicated in the control of aggression and/or impulsivity in humans and been involved in suicide. This gene has a functional polymorphism in which there is a variable number tandem repeat (VNTR) in the upstream region (MAOA-uVNTR). We hypothesized that MAOA dysfunction due to this polymorphism was associated with suicide genetically through the disinhibition of aggression and/or impulsivity. We performed an association study between completed suicides and the MAOA-uVNTR polymorphism. No significant difference in genotype distribution or allele frequencies was found between completed suicides and comparison groups either in males or females. These results show no evidence of an association between the MAOA-uVNTR polymorphism and completed suicides and suggest that MAOA is not involved in the susceptibility to suicide. PMID- 11920861 TI - Is there a relationship between Wolfram syndrome carrier status and suicide? AB - Wolfram syndrome (WFS) is a rare, autosomal recessive neurodegenerative disorder. An increased risk of psychiatric disorders and suicide has been reported for heterozygote carriers. In this study we investigated whether mutations in the WFS gene are associated with suicide in the general population. The gene for WFS (WFS1) has recently been mapped to chromosome 4p16.1, and its genomic structure has been characterized. We screened the entire WFS1 ORF in a panel of 100 completed suicides, 60 blood donors not known to have psychiatric illness, and 100 donors with a negative history of depression or suicidal behavior. We did not find evidence of an increased incidence of WFS carriers in the suicide panel and concluded that WFS1 carrier status is not a significant contributor to suicide in the general population. Screening of this highly polymorphic gene resulted in the detection of 33 variants, 13 of which cause amino acid changes. Seven of these changes have not been previously reported and six were unique to our suicide panel. PMID- 11920862 TI - Exploratory factor analysis of obsessive-compulsive patients and association with 5-HTTLPR polymorphism. AB - The determination of a genetic basis for obsessive-compulsive disorder (OCD) depends on how phenotypic boundaries are defined. Although a hypothesis for serotonin dysfunction in OCD has been advanced, no genes specifically responsible for serotonin regulation have as yet been definitively related to the etiology of OCD. The phenotypic variability of OCD could be at the basis of the failure of molecular biology investigations to find any genes involved in the liability to the disorder. Obsessive and compulsive contents can aggregate in OCD patients differently; multifactorial description may therefore be able to account for OCD phenotypic variance. Using principal component analysis, we derived five factors from 13 main contents of the Yale-Brown Obsessive-Compulsive Scale (YBOCS), and considered them as quantitative phenotypes to evaluate their possible association with an insertion/deletion polymorphism in the promoter region of the serotonin transporter gene (5-HTTLPR). A trend toward positive association between the fifth factor, including counting and repeating rituals, and 5-HTTLPR was found. However, only considering the subgroup of patients with tic codiagnosis, we found a significantly higher score for the fifth factor for patients with L/L genotype with respect to L/S and S/S genotypes. PMID- 11920863 TI - Sequencing of the GRIK1 gene in patients with juvenile absence epilepsy does not reveal mutations affecting receptor structure. AB - Hereditary factors play a major role in the etiology of juvenile absence epilepsy (JAE) that is a common subtype of idiopathic generalized epilepsy (IGE). Sander et al. [1997: Am J Med Genet 74:416-421] reported an allelic association of JAE with the nine-copy allele of a tetranucleotide repeat polymorphism in the third intron of the kainate-selective GluR5 receptor gene (GRIK1) and supportive evidence for linkage of IGE to GRIK1 in families of JAE probands. These findings suggest that a major genetic determinant of GRIK1 confers susceptibility to JAE. Assuming that the GRIK1 tetranucleotide repeat polymorphism is unlikely to have functional relevance itself, we have sequenced the coding regions and regulatory sequences of the GRIK1 gene in eight JAE patients who carry the nine-repeat allele of the GRIK1 tetranucleotide repeat polymorphism to detect a putative functional GRIK1 mutation that is in linkage disequilibrium with the nine-repeat allele. Seven of them were derived from families showing positive evidence for linkage to GRIK1. Our mutation analysis of coding regions and splice junctions revealed only two silent polymorphisms (A522C and C1173T) out of the five SNPs present in public databases and no mutations affecting protein structure. No significant differences were found in the allele frequencies of the detected polymorphisms between the JAE patients and controls. High levels of sequence conservation were also found in the promoter, in the 5' and both the 3' untranslated regions and in the RNA secondary structure involved in the editing reaction. The results presented indicate that mutations in the coding sequences, in the intron-exon boundaries and in the main regulatory regions of the GRIK1 are not commonly involved in the etiology of JAE. PMID- 11920866 TI - Applications of on-line weak affinity interactions in free solution capillary electrophoresis. AB - The impressive selectivity offered by capillary electrophoresis can in some cases be further increased when ligands or additives that engage in weak affinity interactions with one or more of the separated analytes are added to the electrophoresis buffer. This on-line affinity capillary electrophoresis approach is feasible when the migration of complexed molecules is different from the migration of free molecules and when separation conditions are nondenaturing. In this review, we focus on applying weak interactions as tools to enhance the separation of closely related molecules, e.g., drug enantiomers and on using capillary electrophoresis to characterize such interactions quantitatively. We describe the equations for binding isotherms, illustrate how selectivity can be manipulated by varying the additive concentrations, and show how the methods may be used to estimate binding constants. On-line affinity capillary electrophoresis methods are especially valuable for enantiomeric separations and for functional characterization of the contents of biological samples that are only available in minute quantities. PMID- 11920867 TI - Use of bioaffinity interactions in electrokinetically controlled assays on microfabricated devices. AB - In this contribution, the role of bioaffinity interactions on electrokinetically controlled microfabricated devices is reviewed. Interesting applications reported in the literature include enzymatic assays, where enzyme and enzyme inhibition kinetics were studied, often in combination with electrophoretic separation. Attention is paid towards developments that could lead to implementation of electrokinetically controlled microdevices in high-throughput screening. Furthermore, enzyme-facilitated detection in combination with electrophoretic separation on microdevices is discussed. Various types of immunoassays have been implemented on the microchip format. The selectivity of antibody-antigen interaction has been exploited for the detection of analytes in complex sample matrices as required, for example, in clinical chemistry. Binding kinetics as well as stoichiometry were studied in chip-based assays. Automated mixing protocols as well as the demonstration of a parallel immunoassay allow implementation of microdevices in high-throughput screening. Furthermore, demonstration of immunoassays on cheap polymeric microdevices opens the way towards the fabrication of disposable devices, a requirement for commercialization and therefore for application in routine analyses. PMID- 11920868 TI - Immunoaffinity capillary electrophoresis: determination of binding constant and stoichiometry for antibody-antigen interaction. AB - Affinity capillary electrophoresis (ACE) can provide both qualitative and quantitative information on molecular interactions and affords the advantages of very low sample consumption, high mass sensitivity, short analysis time, and the use of automated instrumentation. It has been applied clinically and biochemically to the determination of the binding constant and to the measurement of the binding stoichiometry for interactions between antibodies (Ab's) and antigens (Ag's) in free solution. In many situations, the Ag molecule has two or multiple binding sites, each of which has a similar or different intrinsic affinity for binding independently to the combining site(s) on an Ab molecule. The multivalent binding reactions between Ab and Ag molecules often occur. The objective of this review is to describe the uses of ACE in the determination of binding constants and stoichiometry of Ab-Ag interactions (immunoaffnity capillary electrophoresis), focusing especially on multivalent Ab-Ag interaction modes. Five model binding systems developed recently using ACE techniques are described with principles and examples: (i) divalent mAb-monovalent Ag interaction, (ii) divalent mAb-(homo)polyvalent Ag interaction, (iii) cooperativity of two binding sites of mAb-monovalent Ag interaction, (iv) monovalent Fab-divalent Ag interaction, and (v) polyclonal Ab-monovalent Ag interaction. Finally, the determination of binding stoichiometry of Ab-Ag interactions by ACE is described. PMID- 11920869 TI - Evaluation of molecule-microbe interactions with capillary electrophoresis: procedures, utility and restrictions. AB - Understanding the interactions between molecules and living organisms is of paramount importance for the evaluation of pharmaceutical activity, chemical toxicity and all manner of microbiological studies. The capability of capillary electrophoresis (CE) in the evaluation of molecule-microbe interactions is examined in the present paper. The fundamental chemical concept of the binding or association constant for molecular systems measured in free solution is discussed for biological systems where microorganisms uptake or associate with molecules from their environment. The heterogeneity of the living organisms must be understood and accounted for including differences related to semantics such as concentration units and the nature of the associations between two entities and large differences in the size and number of microorganisms as compared to molecules. Finally, the added complexity and even inhomogeneity of a cell compared to most molecular systems must be considered and possibly controlled. The binding of specific molecules to viruses is discussed. CE can be utilized to quickly determine if a molecule binds very strongly or not at all to a cell (i.e., a binary yes/no answer). This could be useful for initial high-throughput screening purposes when using capillary arrays, for example. CE can be useful for determining unusual (large) molecule/microbe stoichiometries. Finally, CE can sometimes be used to determine the size of binding constants (K(RL)) within certain limits provided experimental conditions can be formulated that minimize problems of biological heterogeneity. PMID- 11920870 TI - Polymer microfluidic chips for electrochemical and biochemical analyses. AB - Our recent developments concerning the fabrication of polymer microchips and their applications for biochemical analyses are reviewed. We first describe two methods of fabrication of polymer microfluidic chips, namely UV-laser photoablation and plasma etching that are well suited for the prototyping and mass fabrication of microchannel networks with integrated microelectrodes. These microanalytical systems can be coupled with various detection means including mass spectrometry, and their applications in capillary electrophoresis are presented here. We also present how UV laser photoablation can be used for the patterning of biomolecules on polymer surfaces for generating two-dimensional arrays of microspots to carry out affinity assays. Finally, the use of the microchips for the development of fast affinity and immunological assays with electrochemical detection is presented, demonstrating the potential of these polymer microchips for medical diagnostics and drug discovery. PMID- 11920871 TI - Molecular interactions of glycopeptide antibiotics investigated by affinity capillary electrophoresis and bioaffinity electrospray ionization-mass spectrometry. AB - Many analytical approaches are available to evaluate (bio)molecular interactions, all of which have their particular advantages and disadvantages. In recent years, two relatively new techniques have emerged that may be used by the bioanalytical community to evaluate such interactions, namely affinity capillary electrophoresis (ACE) and bioaffinity electrospray ionization-mass spectrometry (ESI-MS). In this paper, we describe and evaluate the use of both these techniques for the investigation of the interactions of glycopeptide antibiotics with peptides that mimic the bacterial cell wall binding site. We focus particularly on the effect of the sugar moieties attached to the antibiotic peptide backbone and on the noncovalent dimerization of these glycopeptide antibiotics. PMID- 11920872 TI - Accurately describing weak analyte-additive interactions by capillary electrophoresis. AB - When modeling analyte-additive interactions in capillary electrophoresis (CE), it is necessary to correct for all changes in the apparent electrophoretic mobility of an analyte that are not due to specific binding. Current models based on dynamic complexation have corrected for bulk viscosity changes in the background electrolyte (BGE) when additives are used, while assuming negligible changes in the dielectric constant and other physicochemical properties of the solution. In this report, a study of weak interactions between deoxyribonucleotides and hydroxypropyl-beta-cyclodextrin (HP-beta-CD) revealed significant nonideality in binding isotherms. Changes in the dielectric properties of the solution due to the addition of high concentrations of HP-beta-CD to the BGE was observed to alter the electrophoretic mobility of analytes. A relative dielectric correction factor was required to normalize analyte mobilities to a reference state of zero additive concentration. The use of both a relative dielectric factor and a viscosity correction factor was found to increase the accuracy of the model, reflected by a higher degree of correlation between predicted and measured analyte mobilities. This type of correction is particularly relevant when studying weak analyte binding interactions or when using high concentrations of additive in the BGE. This work is vital for accurate determination of weak binding constants and mobility values, as well as providing a deeper understanding of the fundamental parameters influencing a separation in CE. PMID- 11920873 TI - Computer simulation of affinity capillary electrophoresis. AB - A computer-simulated model of affinity capillary electrophoresis is developed. Unlike existing models, it is able to describe the situation where the concentrations of sample molecules and ligand molecules are commensurable, or even the situation where the zones occupied by these molecules are not mixed initially. The model permits to study the dependence of the spatial and temporal distributions of sample molecules on various parameters such as reaction rate constants, concentrations of sample and reagent, electromigration velocities of sample and reagent and sample injection volume. A collection of peak shapes for different values of parameters is presented. The dependence of peak variance on the ratio of the time of analysis to the characteristic time of reaction is studied. PMID- 11920874 TI - Kinetics of thermal denaturation of human rhinoviruses in the presence of anti viral capsid binders analyzed by capillary electrophoresis. AB - In vivo, the icosahedral capsid of human rhinoviruses undergoes well-defined transitions during the infection pathway. Native virus, sedimenting at 150S, is converted to subviral particles with a sedimentation coefficient of 135S, which have lost the innermost capsid protein VP4. Upon release of the genomic RNA empty 80S capsids remain. Similar structural modifications are observed in vitro upon exposure to low pH and/or elevated temperature. Virions are stabilized against these transitions by various antiviral compounds, which bind to a hydrophobic pocket in the capsid protein VP1. Using capillary electrophoresis the kinetics of viral decay in the presence of such hydrophobic drugs was investigated. Assuming first-order kinetics, the increase of the time constant reflects the extent of stabilization. Exposure of the virions to 55 degrees C after presaturation with the antivirals increased the time constants (as compared to native virus) by a factor of 8-30, from a few minutes to several ten minutes. Denaturation of the stabilized capsid gave rise to heterogeneous material rather than to defined subviral particles. This was confirmed by electron microscopy and indicates that the structural modification of the virus follows a kinetically well-defined pathway which is disturbed by the drugs resulting in disorganized disruption of the virion. PMID- 11920875 TI - Fluorescence polarization detection for affinity capillary electrophoresis. AB - Affinity capillary electrophoresis (ACE) with laser-induced fluorescence polarization (LIFP) detection is described, with examples of affinity interaction studies. Because fluorescence polarization is sensitive to changes in the rotational motion arising from molecular association or dissociation, ACE-LIFP is capable of providing information on the formation of affinity complexes prior to or during CE separation. Unbound, small fluorescent probes generally have little fluorescence polarization because of rapid rotation of the molecule in solution. When the small fluorescent probe is bound to a larger affinity agent, such as an antibody, the fluorescence polarization (and anisotropy) increases due to slower motion of the much larger complex molecule in the solution. Fluorescence polarization results are obtained by simultaneously measuring fluorescence intensities of vertical and horizontal polarization planes. Applications of CE LIFP to both strong and weak binding systems are discussed with antibody-antigen and DNA-protein binding as examples. For strong affinity binding, such as between cyclosporine and its antibody, complexes are formed prior to CE-LIFP analysis. For weaker binding, such as between single-stranded DNA and its binding protein, the single-stranded DNA binding protein is added to the CE separation buffer to enhance dynamic formation of affinity complexes. Both fluorescence polarization (and anisotropy) and mobility shift results are complementary and are useful for immunoassays and binding studies. PMID- 11920876 TI - Immunoassay of serum alpha(1)-antitrypsin by affinity-probe capillary isoelectric focusing using a fluorescence-labeled recombinant antibody fragment. AB - An immunoassay for human alpha(1)-antitrypsin based on affinity-probe capillary isoelectric focusing (AP-CIEF) is described. The method is based on the separation of free and bound labeled antibody fragments by CIEF with laser induced fluorescence detection. A recombinant Fab' fragment of mouse immunoglobulin G1 (IgG1) against human alpha(1)-antitrypsin was labeled with tetramethylrhodamine on the single cysteine residue at the hinge region. A single pI isomer of the labeled Fab' was purified by IEF in a slab of agarose gel and was then used as the affinity probe for alpha(1)-antitrypsin. The use of recombinant Fab' considerably simplified the labeling process. Although there was some difficulty in the quantification of native alpha(1)-antitrypsin with the affinity probe, carbamylation of the antigen sample by heat treatment with urea made the complex peaks appear reproducibly and more distinct, thus facilitating the identification and quantification of the complex. The system provided an almost linear response to a pure sample of alpha(1)-antitrypsin over a concentration range of 5-1000 ng/mL and the detection limit extended down to around 2 ng/mL. Alpha(1)-antitrypsin in a serum sample was determined using this system to be 1.2 mg/mL which is comparable to the reported value for the protein. PMID- 11920877 TI - Capillary electrophoresis investigation of a partially unfolded conformation of beta(2)-microglobulin. AB - Dialysis-related amyloidosis is a disease in which partial unfolding of beta(2) microglobulin plays a key pathogenetic role in the formation of the amyloid fibrils. We have recently demonstrated that a partially unfolded conformer of beta(2)-microglobulin is involved in fibrillogenesis and that this species is significantly populated under physiological conditions. In this work capillary electrophoresis has been used to measure the equilibrium between the native protein and this conformer in samples known to have a higher or lower amyloidogenic potential, namely full-length beta(2)-microglobulin, two truncated species and a mutant, created by replacing histidine in position 31 with thyrosine. In addition, for all protein species folding stability experiments have been carried out by monitoring the secondary structure by circular dichroism at increasing concentrations of guanidinium chloride. The values of free energy of unfolding in the absence of denaturant, obtained by elaboration of these experiments, were found to be inversely correlated to the area percent of the partially unfolded conformer, as measured by capillary electrophoresis. Affinity capillary electrophoresis experiments have been also carried out under nondenaturing conditions to assess the affinity of copper and suramin to either the native form or the conformational intermediate of full-length beta(2) microglobulin. PMID- 11920878 TI - Use of peptide nucleic acid probes for detecting DNA single-base mutations by capillary electrophoresis. AB - Peptide nucleic acid (PNA) oligomers can be used as probes in pre-gel hybridization experiments, as an alternative to Southern hybridization. In this technique, the PNA probe is hybridized to a cyanine-5 labeled DNA sample denatured at low ionic strength, and the mixture is directly injected for size separation into a capillary electrophoresis (CE) system equipped with laser induced fluorescence (LIF) detector. The neutral backbone of PNA allows hybridization to occur at low ionic strength and assures an efficient CE separation of the PNA/DNA hybrids from both double-stranded and single-stranded DNA. We have used as a model system the cystic fibrosis R553X and R1162X single base mutations and we have assessed the influence of various factors, such as temperature and denaturants concentration on DNA/PNA hybrid stability in order to achieve the high specificity required for a single base pair discrimination. PMID- 11920879 TI - Detection of unwanted protein-bound ligands by capillary zone electrophoresis: the case of hidden ligands that stabilize cholinesterase conformation. AB - Detection, identification and characterization of compounds present in purified proteins and biopharmaceuticals are of central interest. As well as chemical remedies, proteins of pharmacological interest have to exhibit their nakedness to become therapeutic drugs. Cholinesterases (ChE) are enzymes of major importance for detoxification of poisonous esters. Likewise, ChE are characterized by the high catalytic efficiency of an active site positioned at the bottom of a deep gorge. The gorge can be partially or fully occupied by ligands, i.e., substrates and inhibitors that are currently used in affinity chromatography purification steps. Accordingly, a suitable method allowing to analyse the presence of unwanted ligands and its influence on the functional conformation and stability of these enzymes was essential. We have developed CZE approaches for that purpose. The factors causing discrepancies between data for thermal unfolding of ChE by electrophoretic and by calorimetric methods were investigated. The presence of unwanted hidden ligands bound to purified enzymes was first demonstrated. The incidence of these ligands was discussed. Altogether, our results raised several questions concerning the real conformation of the native state of enzymes. Finally, CZE was proved to be a pertinent tool to validate the conformity of purified enzymes to a status of biopharmaceutical. PMID- 11920880 TI - A study of the binding between polymers and peptides, using affinity capillary electrophoresis, applied to polymeric drug delivery systems. AB - We have investigated the potential of affinity capillary electrophoresis (ACE) to evaluate binding constants between an anionic polydispersed polymer and four peptides. Nonlinear regression and three current linearization methods, the y reciprocal, the x-reciprocal and the double-reciprocal, were employed for the estimation of the binding constants. The x-reciprocal and the double-reciprocal plots indicated the presence of two portions of straight lines for angiopeptin, triptorelin and the thyrotropin releasing hormone (TRH), and therefore the probable existence of a second-order interaction which causes the deviation from the 1:1 model. Peptide 1 exhibited a unique binding constant of 2.4 x 10(6)M(-1). In contrast, angiopeptin, triptorelin and TRH exhibited a K(1) of 4.0 x 10(6), 5.3 x 10(6) and 20.2 x 10(6)M(-1), respectively, and a K(2) of 0.4 x 10(6), 0.5 x 10(6) and 1.4 x 10(6)M(-1), respectively. The origin of the high scattering of the data points was further investigated. Neither the viscosity, nor the adsorption of the peptides to the capillary wall appeared to be the determining factor of data scattering. Finally, a possible adsorption of the polymer leading to the electroosmotic flow instability was supposed. PMID- 11920881 TI - Application of affinity capillary electrophoresis for the determination of binding and thermodynamic constants of enediynes with bovine serum albumin. AB - The binding constants and thermodynamic properties of a series of novel enediyne compounds with bovine serum albumin (BSA) were determined. The enediynes were synthesized, characterized, and then studied by affinity capillary electrophoresis (ACE) methods to derive these recognition parameters. Change in electrophoretic mobility of BSA as a function of enediyne concentration was determined at 25 degrees C providing binding constants of 1.76 x 10(5), 1.14 x 10(5), and 0.68 x 10(5) M(-1) for enediynephenylalanine carboxylic acid, enediynephenylalanine methyl ester, and enediyne carboxylic acid, respectively. The binding constant for the enediynephenylalanine carboxylic acid was in good agreement with that obtained using conventional methodology. Binding constants for the interaction of enediynes with BSA decreased with an increase in temperature. Van't Hoff plots showed a direct correlation between intensity of the binding constant and the sign and magnitude of various thermodynamic parameters (DeltaG, DeltaS, and/or DeltaH). PMID- 11920882 TI - Drug-plasma protein binding assay by electrokinetic chromatography-frontal analysis. AB - We developed a rapid, microscale and reliable analytical method for binding of drugs to plasma proteins using capillary electrophoresis (CE) with ionic cyclodextrins (CD) combined with frontal analysis. These CDs were used as pseudostationary phases of electrokinetic chromatography (EKC). The CD-modified EKC (CDEKC) approach allowed us to separate anionic drugs from plasma proteins, whereas CZE could not separate these drugs from plasma proteins because they had a similar mobility like plasma proteins. CDs uniquely interact with these drugs but not with plasma proteins. Therefore, CDEKC could be coupled with frontal analysis to measure the binding of anionic drugs to plasma proteins. The binding values obtained by CDEKC were highly consistent with those determined by the ultrafiltration method. Our CDEKC approach should expand the applicability of CE to protein binding analysis. PMID- 11920883 TI - Identification of drug-binding sites on human serum albumin using affinity capillary electrophoresis and chemically modified proteins as buffer additives. AB - A technique based on affinity capillary electrophoresis (ACE) and chemically modified proteins was used to screen the binding sites of various drugs on human serum albumin (HSA). This involved using HSA as a buffer additive, following the site-selective modification of this protein at two residues (tryptophan 214 or tyrosine 411) located in its major binding regions. The migration times of four compounds (warfarin, ibuprofen, suprofen and flurbiprofen) were measured in the presence of normal or modified HSA. These times were then compared and the mobility shifts observed with the modified proteins were used to identify the binding regions of each injected solute on HSA. Items considered in optimizing this assay included the concentration of protein placed into the running buffer, the reagents used to modify HSA, and the use of dextran as a secondary additive to adjust protein mobility. The results of this method showed good agreement with those of previous reports. The advantages and disadvantages of this approach are examined, as well as its possible extension to other solutes. PMID- 11920884 TI - Mapping of stereoselective recognition sites on human serum transferrin by capillary electrophoresis and molecular modelling. AB - Stereoselective recognition of chiral compounds can be used for mapping of surface interaction sites on proteins. Iron-free human serum transferrin is a suitable chiral selector in capillary electrophoresis used in native form in solution. Separation of optical isomers of tryptophan-methylester, tryptophan ethylester and tryptophan-butylester and various drugs were studied in capillary zone electrophoresis applying a distinct transferrin zone prior to sample injection. Changes in the electrophoretic patterns (i.e., in the migration properties) of the molecules reflected the possible interactions with the protein. The tryptophan derivatives and eight drugs possessed stereoselective interactions, seven drugs showed interactions without appreciable chiral separation, and the others did not present any direct complexation with the protein molecules. Molecular modelling was performed to characterize the binding areas at the iron binding site of iron-free transferrin. The docking of tryptophan derivatives on transferrin showed that the R-enantiomers possess a stronger complexation with transferrin, whereas the S-enantiomers are bound by weaker interactions, which is in excellent agreement with the capillary electrophoresis results, where the R-enantiomers were always retarded stronger by transferrin. A ranking of drugs by the lipo score parameter of the docking shows an accordance with the stereoselective interactions by the protein. PMID- 11920887 TI - Editorial. PMID- 11920885 TI - Chiral separation of gemifloxacin in sodium-containing media using chiral crown ether as a chiral selector by capillary and microchip electrophoresis. AB - Chiral crown ether, (+)-(18-crown-6)-tetracarboxylic acid (18C6H(4)), is an effective chiral selector for resolving enantiomeric primary amines owing to the difference in affinities between 18C6H(4) and each of the amine enantiomers. In addition to the destacking effect of sodium ion in the sample solution, the strong affinity of sodium ion to the polyether ring of crown ether is unfavorable to chiral capillary electrophoresis using 18C6H(4) as a chiral selector. In this report, the chiral separation of gemifloxacin dissolved in a saline sample matrix using 18C6H(4) was investigated. Adding a chelating agent, ethylenediaminetetraacetic acid (EDTA), to the run buffer greatly improved the separation efficiencies and peak shapes. The successful chiral separation of gemifloxacin in a urinary solution was demonstrated for both capillary and microchip electrophoresis. PMID- 11920888 TI - Correlation of ultrasound findings and biochemical markers in the second trimester of pregnancy in fetuses with trisomy 21. AB - OBJECTIVE: The aim of the present study was to assess possible correlations between ultrasound findings and maternal serum biochemical ('triple test') markers among fetuses with trisomy 21 in the second trimester of pregnancy. METHODS: The study was a retrospective cohort study of 72 pregnancies affected by trisomy 21 who had a second trimester ultrasound and biochemical screen performed at a single center between 1990 and 1999. The biochemical screen consisted of alpha-fetoprotein (AFP), total beta human chorionic gonadotrophin (hCG) and estriol (uE(3)). Marker levels were expressed in multiples of the median (MoM). The ultrasound findings assessed were major structural anomalies, short humerus length, short femur length, increased nuchal fold thickness (NF), hyperechoic bowel, echogenic intracardiac focus (EIF), ventriculomegaly, choroid plexus cysts and renal pyelectasis. RESULTS: Second trimester maternal serum biochemical markers and ultrasound findings appeared to be largely independent of each other. However, some significant correlations were observed. Estriol was significantly lower when a fetal cystic hygroma was detected on ultrasound compared to those with no cystic hygroma (0.40 vs. 0.70 MoM, p<0.05). The median hCG level was significantly lower in those pregnancies with a normal second trimester fetal ultrasound compared to those with positive ultrasound findings (2.07 vs. 2.87 MoM, p<0.05). Median hCG levels were also significantly higher in those cases with NF> or =5 mm as compared to those with NF<5 mm (2.99 vs. 2.49 MoM, p<0.05). This difference persisted after exclusion of the five cases with cystic hygromas (2.99 vs. 2.49 MoM, p<0.05). A significant positive correlation was observed between log(10) hCG and log(10) NF MoM (Spearman's rho=0.252, p<0.05). NF was significantly greater among fetuses with an identifiable cardiac defect compared with those without a detectable cardiac defect (median of 7.0 mm vs. 3.8 mm, p<0.01). This difference persisted when expressed as multiples of the median (2.8 vs. 1.3 MoM, p<0.01). CONCLUSION: Second trimester ultrasound and biochemical markers are largely independent in fetuses with trisomy 21, however significant correlations between the two were observed in the present series. These may be important in screening protocols that combine second trimester ultrasound and biochemical markers. PMID- 11920889 TI - Maternal serum human chorionic gonadotrophin and pregnancy-associated plasma protein A in twin pregnancies in the first trimester. AB - OBJECTIVES: To determine the levels of free beta-human chorionic gonadotrophin (beta-hCG) and pregnancy-associated plasma protein A (PAPP-A) in twin pregnancies in the first trimester. METHODS: Serum samples were obtained from 67 pregnant women with twin pregnancies and maternal serum free beta-hCG and PAPP-A concentrations were compared with those of 4279 singleton controls between the 8th and 13th weeks of gestation. RESULTS: The geometric means of chromosomally normal twin pregnancies were 1.85 MoM for free beta-hCG and 2.36 MoM for PAPP-A. There were no cases affected by Down syndrome in either group. CONCLUSION: Twin pregnancies secrete more PAPP-A than expected on the basis of singleton controls whereas free beta-hCG production is not increased. The results of the present study can be used to establish normal reference values when introducing first trimester Down syndrome screening in prenatal care. PMID- 11920890 TI - Single umbilical artery stenosis associated with intrauterine fetal death post transfusion. AB - Single umbilical artery is among the most common funicular vascular anomalies. In contrast, umbilical artery stenosis is rare, and has only been reported in three vessel cords. We describe a case of single umbilical artery stenosis in a fetus with no associated malformations. Intrauterine fetal death occurred at 28 weeks' gestation following cordocentesis and intravascular transfusion for Rhesus alloimmunization. Single umbilical artery stenosis may place the fetus at increased risk, particularly in cases requiring interventions involving cord manipulation. PMID- 11920891 TI - Hyperechogenic fetal bowel and Down syndrome. Results of a French collaborative study based on 680 prospective cases. AB - Hyperechogenic fetal bowel is prenatally detected by ultrasound during the second trimester of pregnancy in 0.1% to 1.8% of foetuses. It has been described as a normal variant and has often been associated with severe diseases, notably Down syndrome. The aim of the present study was to determine the risk of trisomy 21 in a prospective study of 680 fetuses with hyperechogenic foetal bowel. Karyotyping was performed on amniotic cells in 632 cases, and outcome was known in 655 cases. A 2.5% risk of Down syndrome and a 1% risk of other severe chromosomal anomalies were observed. Hyperechogenicity was isolated in 11/17 Down syndrome cases, and associated with other ultrasound anomalies in all seven cases of severe chromosomal anomalies. In conclusion, fetal bowel hyperechogenicity indicates a risk of chromosomal anomalies ten-fold higher than that expected on the basis of maternal age, therefore justifying invasive procedures. PMID- 11920892 TI - Rapid interphase analysis for prenatal diagnosis of translocation carriers using subtelomeric probes. AB - Interphase fluorescence in situ hybridization (FISH) has become an accepted laboratory technique for the rapid and preliminary prenatal assessment of chromosome aneuploidy. The introduction of subtelomeric FISH probes now allows for the molecular-cytogenetic analysis of terminal chromosome rearrangements. In a prospective study, we examined the prenatal use of subtelomeric probes on interphase cells to rapidly detect the carrier status of a fetus when a parent carried a known reciprocal or Robertsonian chromosome translocation. Three of the cases were identified as being abnormal. All cases were confirmed by routine cytogenetic analysis. These findings clearly demonstrated the utility of this technique and these probes to rapidly and correctly identify balanced and unbalanced chromosome anomalies of a fetus that could result from parental translocations. PMID- 11920893 TI - Prenatal diagnosis of long QT syndrome using magnetocardiography: a case report and review of the literature. AB - OBJECTIVES: To investigate the usefulness of magnetocardiography (MCG) in the prenatal diagnosis of fetal long QT syndrome. METHODS: Fetal MCG was recorded in a case of fetal long QT syndrome suspected in utero. The literature on the prenatal diagnosis of fetal long QT syndrome was also reviewed. RESULTS: The MCG was performed at 36 weeks' gestation because sustained fetal bradycardia of 110 120 bpm was detected by cardiotocography. The 64-channel MCG revealed a prolonged fetal corrected QT-interval of 0.57 s. The postnatal electrocardiogram coincided with prenatal MCG. CONCLUSION: An accumulation of cases of prenatally diagnosed long QT syndrome using MCG indicates that MCG may be the most reliable tool for the prenatal diagnosis of long QT syndrome. PMID- 11920894 TI - Factors influencing the flow rate through a surgical defect in human fetal membranes. AB - OBJECTIVES: In order to determine factors influencing the flow rate trough a created defect in human fetal membranes, an ex vivo set-up was used with fetal membranes collected from patients undergoing Caesarean section at term. METHODS: The membranes were secured at the bottom of a plastic tube and traumatised with needles ranging from 14-26 Gauges (Ga), under a hydrostatic pressure of 10 to 20 cm H(2)O and an angle of 45 degrees or 90 degrees. The column was filled with amniotic fluid or Hartmann's solution. The duration of the puncture was 1 s or the time it takes to aspirate 10 ml through the needle. The flow rate through the defect in the fetal membranes and size of the defect were measured. RESULTS: The flow rate and defect size increased with increasing diameter of the needle. Increasing the pressure in the column resulted in a significant linear increase in the flow rate. Replacing the saline solution with amniotic fluid did not result in significant changes in the measured flow rates, except for the small needle size (24 Ga). Increasing the duration of the puncture did not result in increased flow rates, except for small needle size (24 Ga). CONCLUSION: These experiments suggest that needle diameter, angle of needle insertion, duration of the procedure, amniotic fluid pressure and composition could influence the incidence of amniotic fluid leakage following amniocentesis. PMID- 11920895 TI - Obstetric outcome after fetal reduction to singleton pregnancies. AB - OBJECTIVE: To study the outcome after fetal reduction or selective termination to singleton pregnancies for various indications. METHODS: Fetal reduction or selective feticide to singleton pregnancies was performed in 80 multiple gestations (congenital malformations, 17 cases; high-risk obstetric conditions, 25 cases; or social/psychological indications, 38 cases). RESULTS: The overall pregnancy loss rate was 10%; however, pregnancy failure was significantly higher in selective reductions performed for preterm prelabor rupture of membranes (PPROM) (4/8) compared with monochorionic twin and bad obstetric history. Fetal reduction to singletons for psychological reasons resulted in a pregnancy wastage of 5.3% (2/38). Procedures performed at < or =14 weeks showed a significantly lower fetal loss rate (2/61; 3.3%), a higher mean gestational age at delivery (38.3+/-2.2 weeks), and a decreased prematurity rate (p< or =0.001). The number of reduced fetuses, prenatal diagnosis by chorionic villus sampling before the reduction and maternal age did not interfere with pregnancy outcome. CONCLUSION: Fetal reduction to singleton pregnancies has a favorable outcome, especially when performed before 14 weeks of gestation. PMID- 11920896 TI - The first trimester 'combined test' for the detection of Down syndrome pregnancies in 4939 unselected pregnancies. AB - The high detection rate (DR) for Down syndrome (DS) pregnancies which can be achieved by measuring fetal nuchal translucency (NT) early in pregnancy can be improved by combining it with placental hormones [pregnancy-associated plasma protein A (PAPP-A) and free beta-human chorionic gonadotrophin (fbeta-hCG)] and maternal age ('combined test'). In this study we wanted to assess the DR using the 'combined test' in an unselected population of self-referred pregnant women at a false-positive rate (FPR) of about 5%. NT, PAPP-A, fbeta-hCG and maternal age were measured in all women with singleton pregnancies who booked for delivery in our hospital from 1 December 1997 to 31 April 2000 and who were between 10 and 13 completed weeks of gestation [crown-rump length (CRL) 35-70 mm]. The specific DS risk was calculated using the computer program Alpha Version 5aa (Logical Medical Systems, London, UK). A total of 4939 women were tested. Out of 14 DS pregnancies that occurred during this period of time, 12 were detected with the test. A total of 246 women had a false-positive test result in a non-DS pregnancy (FPR 5.0%). This makes the 'combined test' by far the best test for the detection of DS pregnancies in a low-risk population. The constant increase in maternal age at the time of delivery can also lead to an improved DR if a simple age-dependant protocol for DS detection is used, but only at the price of a much higher number of amniocenteses and subsequent abortions. The DR for DS can be increased much more markedly using the 'combined test' with a FPR that still remains at the level as it was in the early 1970s. PMID- 11920897 TI - Dilemmas encountered by health practitioners offering nuchal translucency screening: a qualitative case study. AB - OBJECTIVE: To explore dilemmas experienced by practitioners involved in routine prenatal nuchal translucency (NT) screening. DESIGN: Qualitative study incorporating multidisciplinary practitioner discussion groups led by a health care ethicist. SETTING: Inner-city teaching hospital with fetal medicine unit. PARTICIPANTS: Thirty-two practitioners whose work relates directly or indirectly to perinatal care. RESULTS: Practitioners identified a number of dilemmas, many of which centred on the tension between pregnancy being seen as a normal or a 'risky' time. Practitioners and women were perceived to have contrasting reasons for screening, with women welcoming the opportunity to 'see' their baby on the ultrasound scan, whilst practitioners were screening for abnormalities. These differing agendas led to various dilemmas particularly in relation to information giving, performing scans incorporating NT screening and promoting individual client choice. CONCLUSIONS: Plans to introduce routine NT screening need careful prospective consideration of the potential implications for both providers and users of the service. The discussion groups also identified the need for training in the complex communication skills required and an awareness of the related ethical dilemmas, plus the need for increased time and resources to enable practitioners to promote informed choice. PMID- 11920898 TI - Maternal serum leptin concentration during the second trimester of pregnancy: association with fetal chromosomal abnormalities. AB - Recent studies suggest that leptin, the product of the obese gene, is produced by the placenta during pregnancy. The present study addressed the question whether second trimester maternal serum leptin could be altered by fetal Down syndrome or Edwards syndrome. Maternal serum leptin concentrations were measured in 18 pregnancies complicated with Down syndrome, six pregnancies complicated with Edwards syndrome and 183 uncomplicated pregnancies during the second trimester of pregnancy. The present results demonstrate that leptin concentrations in uncomplicated pregnancies slightly decrease from the 16th week of pregnancy, reaching a minimum of 18.8 ng/ml around the 20th week, and then rapidly increase to 28.2 ng/ml by the 24th week. Leptin correlation with maternal body weight decreases from r=0.695 at 16-17 week of gestation to r=0.544 at >22 weeks of gestation. There was no significant difference between the mean MoMs of Down syndrome- (0.926) or Edwards syndrome- (0.960) affected pregnancies and normal pregnancies (1.002). A weak correlation (r=0.18, p<0.02) was observed between corrected leptin MoMs and human chorionic gonadotrophin (hCG) MoMs in normal pregnancies. It is assumed that around the 20th week of pregnancy placental leptin production is activated or at least is accelerated and it is added to the amount of leptin produced by maternal adipose tissue. Fetal Down syndrome or Edwards syndrome does not seem to alter maternal leptin concentration and therefore leptin cannot be used as a marker for these chromosomal abnormalities in the early second trimester of pregnancy. PMID- 11920899 TI - Biochemical screening for Down syndrome in pregnancies following renal transplantation. AB - OBJECTIVE: To assess the performance of the double marker test [free beta-human chorionic gonadotrophin (beta-hCG) and alpha-fetoprotein (AFP)] as a screening test for Down syndrome in pregnant patients who had a prior renal transplant. DESIGN: A retrospective study. SETTING: The Fetal Medicine Unit, Royal Free Hospital, London, UK. METHODS: Detailed records of 14 post-renal transplant pregnancies were obtained from the Renal Unit of our hospital where the patients were followed up. The serum concentrations of urea, creatinine, free beta-hCG and AFP at the time of the double marker test were recorded, with a cut-off point of 1:250 for the double marker test. A control group of 14 normal pregnancies matched for age, parity and gestational age was used. The Mann-Whitney U-test and t-tests of unequal variance were applied to compare parameters of the study and the control groups. RESULTS: Two patients in each group were high risk for Down syndrome and amniocentesis revealed normal karyotype. No babies with Down syndrome were delivered in either group. Regression analysis showed significant correlation between free beta-hCG and urea concentrations (p<0.001) and free beta hCG and creatinine concentrations (p<0.001), but not for AFP. CONCLUSIONS: The present study demonstrates that residual renal function alterations persisting after renal transplantation can affect the levels of free beta-hCG and AFP, thus resulting in false-positive screening for Down syndrome. First trimester nuchal translucency (NT) measurement in combination with second trimester ultrasonographic markers can be used in these patients, or alternatively the free beta-hCG levels should be corrected according to the serum creatinine levels. PMID- 11920900 TI - Can we predict 22q11 status of fetuses with tetralogy of Fallot? AB - OBJECTIVE: To determine if chromosome 22q11 deletion status can be predicted in fetuses with tetralogy of Fallot as regards additional phenotypic anomalies. METHODS: One hundred and fifty-one consecutive fetuses with tetralogy of Fallot without or with pulmonary atresia were screened for 22q11 deletion. Additional echographic features [increased nuchal translucency (NT), intrauterine growth retardation (IUGR), polyhydramnios, extracardiac malformations, pulmonary arteries abnormalities] were noted. RESULTS: Twenty-five fetuses had a 22q11 deletion (16.6%). Increased NT, polyhydramnios and IUGR were more frequent in fetuses with 22q11 deletion as well as pulmonary arterial abnormalities. When these different features were present in the same fetus with tetralogy of Fallot, 22q11 deletion can be predicted with a sensitivity of 88%. CONCLUSION: Simple echographic features can help to predict 22q11 status in fetuses with tetralogy of Fallot. This may improve the efficiency of prenatal screening for this defect. PMID- 11920901 TI - Prenatal diagnosis in two families with autosomal, p47(phox)-deficient chronic granulomatous disease due to a novel point mutation in NCF1. AB - OBJECTIVE: Prenatal diagnosis was required in two unrelated families with chronic granulomatous disease (CGD) patients who lacked expression of p47(phox) protein; thus a search for mutations in NCF1 was undertaken. METHODS: Gene scanning was applied to establish the relative number of coding and pseudo-NCF1 genes. PCRs specific for coding NCF1 cDNA and coding NCF1 exon-7 genomic DNA were devised. RESULTS: The normal 1:2 ratio of coding and pseudo-NCF1 genes was found in the patients. Sequencing of the RT-PCR product specific for mRNA from the coding NCF1 genes revealed a novel homozygous G579A mutation in both patients, changing the TGG codon for Trp193 into the TAG stop codon. This mutation was confirmed in genomic DNA. The parents of both patients were found to be heterozygotes for this mutation. In the chorionic villus DNA of the first family a heterozygous G579A mutation was found. Postpartum, functional NADPH oxidase tests were normal. In the second family, this mutation was present in homozygous form in the chorionic villus DNA. Following termination of the pregnancy, the diagnosis of p47(phox) deficient CGD was confirmed on DNA extracted from fetal blood. CONCLUSION: This is the first report of prenatal diagnosis in p47(phox)-deficient CGD. PMID- 11920902 TI - Intrauterine left chamber myocardial infarction of the heart and hydrops fetalis in the recipient fetus due to twin-to-twin transfusion syndrome. AB - A rare complication of twin-to-twin transfusion syndrome (TTTS) is described: myocardial infarction of the recipient fetus. Myocardial infarction and hydrops are considered to be consequences of hypertension in the recipient. No other organs were affected. Pathological signs of intrauterine hypertension were estimated by the thickness of vessel walls and signs of hypertrophied myocardial cells. In the heart of the recipient fetus there was a chronic myocardial infarction near the apex cordis on the anterior wall with an aneurysm 4x5 mm in diameter. Diagnosis was based on light microscopic examination. The poor myocardial systolic function resulted in hydrops. Since the mother was administered beta sympathomimetics in therapeutic doses the contribution of the drug to the myocardial infarction is uncertain, but we would like to suggest this as a possible adverse effect in TTTS. The present case is the first reported myocardial infarction in connection with the syndrome. PMID- 11920903 TI - Accuracy of Down syndrome risks produced in a first-trimester screening programme incorporating fetal nuchal translucency thickness and maternal serum biochemistry. AB - Over the past three years approximately 12 000 women have been screened in the first trimester through our OSCAR programme, which utilizes fetal NT and maternal serum free beta-hCG and PAPP-A. During this time 30 cases of Down syndrome were identified either prenatally or postnatally. Using an established procedure the accuracy of predicted risk for Down syndrome was assessed in a population of 30 cases of Down syndrome and 11 758 unaffected pregnancies. The correlation between predicted risk and prevalence of Down syndrome was very high (r=0.9995). It is concluded that risks produced by the Fetal Medicine Foundation combined risk algorithm agree very closely with Down syndrome prevalence and can be used with confidence when counselling women of their risk. PMID- 11920904 TI - Maternal smoking: age distribution, levels of alpha-fetoprotein and human chorionic gonadotrophin, and effect on detection of Down syndrome pregnancies in second-trimester screening. AB - OBJECTIVES: To study the levels of maternal serum alpha-fetoprotein (AFP) and human chorionic gonadotrophin (hCG) in the second trimester in smokers and non smokers with unaffected and Down syndrome pregnancies; to examine the rate of smoking in different maternal age groups in a population having routine prenatal screening; and to assess the effect of smoking on the detection rates for Down syndrome and corresponding false-positive rates, both overall and in different maternal age groups. METHODS: Information on maternal smoking status, maternal age and serum marker levels was collected from case note searches and the screening programme database on 2272 unaffected singleton pregnancies, 36 unaffected twin pregnancies and 103 singleton Down syndrome pregnancies. RESULTS: In unaffected pregnancies the smokers had a median age 3.3 years less than the non-smokers, while in the Down syndrome cases the corresponding age difference was 2.0 years. Median analyte levels in multiples of the median (MoM) in the unaffected singleton pregnancies were, for non-smokers: AFP=0.97, hCG=1.04; and for smokers, AFP=1.04, hCG=0.80. In the Down syndrome pregnancies the medians were, for non-smokers: AFP=0.69, hCG=2.49; and for smokers, AFP=0.70, hCG=1.53. Correction for smoking status gave median MoMs of 1.0 for both AFP and hCG in the unaffected pregnancies in both smokers and non-smokers. In the Down syndrome cases the corrected medians were, for non-smokers: AFP=0.67, hCG=2.29; and for smokers, AFP=0.73, hCG=1.99. Before correction for maternal smoking the overall detection rate for Down syndrome was 66.7% with a false-positive rate of 6.2%. After correction the detection rate was 67.7% with a false-positive rate of 4.9%. Between the smoking and non-smoking groups there was a significant difference in the detection rate (37.5% versus 76.0%) and the false-positive rate (1.8% versus 8.1%), which disappeared after correction for smoking status (detection rate 62.5% versus 69.3%, false-positive rate 3.9% versus 5.4%). No evidence of a lower incidence of Down syndrome in smokers was found. CONCLUSIONS: While correcting AFP and hCG results for maternal smoking status will have little impact on the overall detection rate for Down syndrome, it may reduce the false-positive rate and will improve the accuracy of the risks given to individual women. PMID- 11920905 TI - The effect of fast reporting by amnio-PCR on anxiety levels in women with positive biochemical screening for Down syndrome--a randomized controlled trial. AB - OBJECTIVE: To study the effect of fast reporting by polymerase chain reaction on amniotic fluid cells (amnio-PCR) on anxiety levels in women with positive biochemical screening for Down syndrome. METHOD: Between May 2000 and April 2001, 60 screen-positive women were randomized before amniocentesis into either having (group A) or not having (group B) fast-reporting by amnio-PCR. Anxiety levels were measured by the Spielberger State-Trait Anxiety Inventory just prior to amniocentesis, three days (when PCR results were known to group A) and three weeks (when standard karyotype results were known to both groups) afterwards. RESULTS: Two women were excluded because in one woman amnio-PCR showed trisomy 21 and the other miscarried shortly after amniocentesis. The state-anxiety scores increased over the three-week period after being informed of the positive-screen result in both groups. The trait- and state-anxiety scores at all points did not differ between the two groups. CONCLUSIONS: In contrast to the general belief, fast reporting by amnio-PCR did not alleviate anxiety in women who are screen positive for Down syndrome. PMID- 11920906 TI - Transcervical chorionic villus sampling in multiple pregnancies using a biopsy forceps. AB - OBJECTIVE: The aim of this study was to assess the effectiveness and safety of chorionic villus sampling (CVS) performed in multiple pregnancies by means of a transcervical biopsy forceps. METHODS: The study included CVS performed from January 1990 to March 2000 in our Unit. The results were analysed in two consecutive periods, period A (1990-1994) and period B (1995-2000), in an attempt to assess the effect of increasing experience. RESULTS: Seventy-five samplings were performed in 39 multiple pregnancies, 38 twin sets and one triplet. A cytogenetic report was obtained in 73% of cases in period A and in 98% in period B. An abnormal karyotype was observed in 11 samples. The need for subsequent amniocentesis decreased from 38% in period A to 10% in period B. The spontaneous fetal loss rate in chromosomally and structurally normal fetuses before the 20th week decreased from 8.7% in period A to 3.3% in period B. The fetal loss rate after the 20th week was 3.3% in period B and none in period A. It must be noted that in three out of the four cases of fetal loss an amniocentesis was needed after CVS. CONCLUSION: Our results suggest that effectiveness and safety improved with increasing experience. Transcervical chorionic villus sampling allows an earlier prenatal genetic diagnosis in multiple pregnancies and this may be particularly relevant for a safer selective termination when chosen by parents if one of the fetuses has an abnormal karyotype. PMID- 11920907 TI - Molecular prenatal diagnosis of non-ketotic hyperglycinemia (glycine encephalopathy). PMID- 11920908 TI - Pitfalls in prenatal and postnatal detection of partial trisomy and tetrasomy 13 by rapid aneuploidy detection. PMID- 11920909 TI - Pregnancy outcomes of women with positive serum screening results for Down syndrome and trisomy 18. PMID- 11920910 TI - Bibliography. Current awareness in prenatal diagnosis. PMID- 11920912 TI - Comparison of the hydrolytic stability of S-(N,N-diethylaminoethyl) isobutyl methylphosphonothiolate with VX in dilute solution. AB - The stability of S-(N,N-diethylaminoethyl) isobutyl methylphosphonothiolate--a V type nerve agent developed by the former Soviet Union--in the environment is an important parameter in threat assessment analysis and for the determination of use, production, testing and storage of this chemical warfare agent. S-(N,N Diethylaminoethyl) isobutyl methylphosphonothiolate is a structural isomer of the nerve agent VX developed by the USA and the UK and will be referred to as VXA (VX analog) in this presentation. Because VXA and VX differ structurally, even though they do have the same molecular formula, it is expected that their physical and chemical properties would be different. This preliminary investigation was undertaken to determine the relative hydrolysis rate of VXA compared with VX. The hydrolysis of each compound at approximately 1 mg x ml(-1) in unbuffered water at pH 7 was determined side-by-side. The half-lives for VXA and VX were determined to be 12.4 days and 4.78 days, respectively. Agent VXA hydrolyzed 2.6 times more slowly than VX, and each agent followed second-order hydrolysis kinetics. These results imply that VXA is more persistent in the environment and therefore poses a greater threat. These results also imply that VXA is more likely to be detected, if present, during an inspection in support of the Chemical Weapons Convention. PMID- 11920913 TI - The active site of human paraoxonase (PON1). AB - Ideally we would like to treat people exposed to nerve agents with an enzyme that rapidly destroys nerve agents. The enzymes considered for such a role include human butyrylcholinesterase (BChE), acetylcholinesterase (AChE), carboxylesterase and paraoxonase (PON1). Success has been achieved in endowing BChE with the ability to hydrolyze organophosphates. The G117H mutant of BCHE hydrolyzes sarin and VX, whereas the double mutant G117H/E197Q hydrolyzes soman (Millard et al. Biochemistry 1995; 34: 15925-15933; 1998; 37: 237-247). However, the rates of organophosphate hydrolysis are slow and a faster organophosphate hydrolase is being sought. Native PON1 hydrolyzes paraoxon with a catalytic efficiency, of 2.4 x 10(6) M(-1) x min(-1), and our goal is to improve the organophosphate hydrolase activity of PON1. To achieve this we need to identify the amino acids in the active site of PON1. Using site-directed mutagenesis and expression in human 293T cells, we have identified the following eight amino acids as being essential to PON1 activity: W280, H114, H133, H154, H242, H284, E52 and D53. Fluorescence of PON1 complexed to terbium ion shows that at least one tryptophan is close to the calcium binding site. PMID- 11920914 TI - Peripheral site ligands accelerate inhibition of acetylcholinesterase by neutral organophosphates. AB - The rates of inhibition of mouse acetylcholinesterase (AChE; EC 3.1.1.7) by paraoxon, haloxon, DDVP and enantiomers of neutral alkyl methylphosphonyl thioates and cationic alkyl methylphosphonyl thiocholines were measured in the presence and absence of AChE peripheral site inhibitors: gallamine, d tubocurarine, propidium, atropine and derivatives of coumarin. All ligands, except the coumarins, at submillimolar concentrations enhanced the rates of inhibition by neutral organophosphates, whereas inhibition rates by cationic organophosphates were decreased. When peripheral site ligand concentrations extended to millimolar concentrations the extent of the enhancement decreased, creating a well-shaped activation profile. Analysis of inhibition by DDVP revealed that peripheral site inhibitors increase the second-order reaction rates by increasing maximal rates of phosphorylation. These observations suggest that peripheral site ligands are capable of allosterically affecting the conformation of residues in the choline binding site of AChE, thus optimizing the position of the leaving group of uncharged organophosphates during the inhibition reaction. PMID- 11920915 TI - Toxicology update: the cardiotoxicity of the oxidative stress metabolites of catecholamines (aminochromes). AB - This toxicology update reviews the oxidative stress metabolites of catecholamines, postulated to be the biochemical initiators of cardiotoxicity. A brief overview of catecholamine metabolism is provided with several noteworthy historical observations relating to the autoxidation and rearrangement of epinephrine. The basic chemical and physical properties of adrenochrome and adrenolutin are discussed. The autoxidative, enzymatic and cellular basis for the transformation of catecholamines to oxidative metabolites is reviewed. Mechanisms seeking to account for the observed cardiotoxic changes in isolated heart perfusion studies and in vivo models are described. PMID- 11920917 TI - Analysis of the degradation compounds of chemical warfare agents using liquid chromatography/mass spectrometry. AB - The analysis of the degradation products of chemical warfare (CW) agents has been a challenge to analysts. The low volatility of these compounds makes them unsuitable for direct gas chromatography analysis without prior derivatization. Lack of a chromophore causes difficulties with classic detection methods after liquid chromatography separation. With the recent development of various interfaces that allow for the introduction of a liquid solvent stream into the mass spectrometer, the task of directly analyzing these compounds has become easier. For this report, we examined three different liquid chromatography/mass spectrometry (LC/MS) interfaces for their suitability for the analysis of CW degradation compounds. The interface types examined were particle beam electron impact ionization (PBI), electrospray ionization (ESI) and atmospheric pressure chemical ionization (APCI). Several alkylphosphonates and thiodiglycol analogs that are produced from the degradation of organophosphorus nerve agents and sulfur mustard, respectively, were analyzed using each of the three techniques. Electron impact ionization following gas chromatography or particle beam introduction typically generates very reproducible, library-searchable mass spectra. Most of the CW breakdown compounds examined using the PBI interface did not produce a molecular ion. Despite the lack of a molecular ion, the mass spectra of the various compounds contained enough different structural information from fragment ions for the positive identification of each. The mass spectra generated using ESI are generally limited to protonated molecular ions with little or no fragmentation. For positive identification and confirmation, tandem mass spectrometry techniques quite often must be used. Many of the compounds in this study were characterized by prominent sodiated adducts along with the protonated molecular ion. Methylphosphonic acid produced protonated dimers, trimers, etc. Although the various adduct ions can be used for additional confirmation of the molecular weight of a compound, the adducts also can result in suppression of ionization of the compound and thus reduce sensitivity. Another 'soft' ionization technique that results in abundant protonated molecular ions is APCI. The mass spectra of the breakdown compounds produced using APCI were characterized generally by either a prominent protonated molecular ion or a dehydrated form of it. In addition, a number of structurally significant fragment ions were observed and their relative abundances could be adjusted by altering the APCI conditions. The data presented here indicate that each of the three techniques can be used successfully for direct liquid introduction and analysis of the non-volatile compounds produced from the degradation of CW agents. The mass spectra produced using each technique are quite different and could be utilized as additional confirmation of compound identity. PMID- 11920916 TI - Development of an immunoassay for diagnosis of exposure to toxic organophosphorus compounds. AB - Currently, diagnosis of exposure to toxic low-molecular-weight compounds is effected by the use of chromatographic techniques. Such an approach is limited by the need for expensive equipment and sample clean-up before carrying out the analysis. To overcome those drawbacks, we have been involved in the development of an immunoassay for diagnosis of exposure to toxic organophosphorus compounds such as pinacolylmethyl phosphonofluoridate (soman), which is a chemical warfare agent. Prior estimates suggested that it is necessary to be able to detect soman at a concentration below 2.5 x 10(-7) M. Using four previously developed monoclonal antibodies, an enzyme-linked immunosorbant assay (ELISA) was used to optimize assay conditions and identify the antibody with the highest apparent affinity. The minimum required assay time was 2.0-2.5 h with no loss in sensitivity. To determine the specificity of the highest affinity antibody, a competitive inhibition enzyme immunoassay (CIEIA) was performed with six structural analogs of soman. The IC50 values for these analogues were 5 x 10(-7) M for 4-nitrophenylpinacolylmethylphosphonate, 8 x 10(-7) M for dipinacolylmethylphosphonate, 2 x 10(-6) M for diisopropylmethylphosphonate, 3 x 10(-5) M for 4-nitrophenylmethyl(phenylphosphinate) and 6.5 x 10(-5) M for 4 nitrophenylethyl(phenyl)phosphinate. 4-Nitrophenyl-di(n-butyl)phosphinate did not inhibit binding. Those inhibitors with branched alkyl side-chains, similar to the soman molecule, were effective inhibitors. Compounds, which contained predominately aromatic groups, were poor inhibitors. We are continuing to probe the binding specificity of the monoclonal antibody to determine its utility in further assay development. Our present results suggest that the antibody chosen may have the appropriate specificity and affinity for immunodiagnosis of exposure to soman. PMID- 11920918 TI - Army medical laboratory telemedicine: role of mass spectrometry in telediagnosis for chemical and biological defense. AB - An army medical field laboratory presently has the capability of performing standard protocols developed at the US Army Medical Research Institute of Chemical Defense for verification of nerve agent or sulfur mustard exposure. The protocols analyze hydrolysis products of chemical warfare agents using gas chromatography/mass spectrometry. Additionally, chemical warfare agents can produce alkylated or phosphorylated proteins following human exposure that have long biological half-lives and can be used as diagnostic biomarkers of chemical agent exposure. An analytical technique known as matrix-assisted laser desorption ionization time-of-flight mass spectrometry (MALDI-TOF/MS) currently is being examined for its potential to analyze these biomarkers. The technique is capable of detecting large biomolecules and modifications made to them. Its fast analysis time makes MALDI-TOF/MS technology suitable for screening casualties from chemical or biological attacks. Basic operation requires minimal training and the instrument has the potential to become field-portable. The limitation of the technique is that the generated data may require considerable expertise from knowledgeable personnel for consultation to ensure correct interpretation. The interaction between research scientists and field personnel in the acquisition of data and its interpretation via advanced digital telecommunication technologies can enhance rapid diagnosis and subsequently improve patient care in remote areas. PMID- 11920919 TI - Progress on the road to new nerve agent treatments. AB - In the 50 years since nerve agents were developed a great deal has been learned about their acute toxicity, treatment and prophylactic strategies. However, the currently fielded treatments are not significantly different from those available at the end of World War II. Reasons for this lack of progress and strategies to circumvent those intrinsic problems that have impeded progress are discussed, with emphasis on the development of scavengers to be introduced as prophylactics that will significantly reduce the effective dose and thus protect against multiple times the normal LD50. PMID- 11920920 TI - The NMDA receptor ion channel: a site for binding of Huperzine A. AB - Huperzine A (HUP-A), first isolated from the Chinese club moss Huperzia serrata, is a potent, reversible and selective inhibitor of acetylcholinesterase (AChE) over butyrylcholinesterase (BChE) (Life Sci. 54: 991-997). Because HUP-A has been shown to penetrate the blood-brain barrier, is more stable than the carbamates used as pretreatments for organophosphate poisoning (OP) and the HUP-A:AChE complex has a longer half-life than other prophylactic sequestering agents, HUP-A has been proposed as a pretreatment drug for nerve agent toxicity by protecting AChE from irreversible OP-induced phosphonylation. More recently (NeuroReport 8: 963-968), pretreatment of embryonic neuronal cultures with HUP-A reduced glutamate-induced cell death and also decreased glutamate-induced calcium mobilization. These results suggest that HUP-A might interfere with and be beneficial for excitatory amino acid overstimulation, such as seen in ischemia, where persistent elevation of internal calcium levels by activation of the N methyl-D-aspartate (NMDA) glutamate subtype receptor is found. We have now investigated the interaction of HUP-A with glutamate receptors. Freshly frozen cortex or synaptic plasma membranes were used, providing 60-90% specific radioligand binding. Huperzine A (< or =100 microM) had no effect on the binding of [3H]glutamate (low- and high-affinity glutamate sites), [3H]MDL 105,519 (NMDA glycine regulatory site), [3H]ifenprodil (NMDA polyamine site) or [3H]CGS 19755 (NMDA antagonist). In contrast with these results, HUP-A non-competitively (Hill slope < 1) inhibited [3H]MK-801 and [3H]TCP binding (co-located NMDA ion channel PCP site) with pseudo K(i) approximately 6 microM. Furthermore, when neuronal cultures were pretreated with HUP-A for 45 min prior to NMDA exposure, HUP-A dose dependently inhibited the NMDA-induced toxicity. Although HUP-A has been implicated to interact with cholinergic receptors, it was without effect at 100 microM on muscarinic (measured by inhibition of [3H]QNB or [3H]NMS binding) or nicotinic [3H]epibatidine binding) receptors; also, HUP-A did not perturb adenosine receptor binding [3H]PIA or [3H]NECA). Therefore, HUP-A most likely attenuates excitatory amino acid toxicity by blocking the NMDA ion channel and subsequent Ca2+ mobilization at or near the PCP and MK-801 ligand sites. Thus, on the one hand, HUP-A could be used as a pretreatment against OPs and it might also be a valuable therapeutic intervention in a variety of acute and chronic disorders by protecting against overstimulation of the excitatory amino acid pathway. By blocking NMDA ion channels without psychotomimetic side-effects, HUP A may protect against diverse neurodegenerative states observed during ischemia or Alzheimer's disease. PMID- 11920921 TI - Combination anticonvulsant treatment of soman-induced seizures. AB - These studies investigated the effectiveness of combination treatment with a benzodiazepine and an anticholinergic drug against soman-induced seizures. The anticholinergic drugs considered were biperiden, scopolamine, trihexaphenidyl, and procyclidine; the benzodiazepines were diazepam and midazolam. Male guinea pigs were implanted surgically with cortical screw electrodes. Electrocorticograms were displayed continually and recorded on a computerized electroencephalographic system. Pyridostigmine (0.026 mg x kg(-1), i.m.) was injected as a pretreatment to inhibit red blood cell acetylcholinesterase by 30 40%. Thirty minutes after pyridostigmine, 2 x LD50 (56 microg x kg(-1)) of soman was injected s.c., followed 1 min later by i.m. treatment with atropine (2 mg x kg(-1)) + 2-PAM (25 mg x kg(-1)). Electrographic seizures occurred in all animals. Anticonvulsant treatment combinations were administered i.m. at 5 or 40 min after seizure onset. Treatment consisted of diazepam or midazolam plus one of the above-mentioned anticholinergic drugs. All doses of the treatment compounds exhibited little or no antiseizure efficacy when given individually. The combination of a benzodiazepine and an anticholinergic drug was effective in terminating soman-induced seizure, whether given 5 or 40 min after seizure onset. The results suggest a strong synergistic effect of combining benzodiazepines with centrally active anticholinergic drugs and support the concept of using an adjunct to supplement diazepam for the treatment of nerve-agent-induced seizures. PMID- 11920922 TI - Beneficial effects of TCP on soman intoxication in guinea pigs: seizures, brain damage and learning behaviour. AB - Poisoning with the potent nerve agent soman produces a cascade of central nervous system (CNS) effects characterized by severe convulsions and eventually death. In animals that survive a soman intoxication, lesions in the amygdala, piriform cortex, hippocampus and thalamus can be observed. In order to examine the mechanisms involved in the effects of soman and to evaluate possible curative interventions, a series of behavioural, electrophysiological and neuropathological experiments were carried out in the guinea pig using the NMDA antagonist N-[1-(2-thienyl)cyclohexyl] piperidine (TCP) in conjunction with atropine and pyridostigmine. The NMDA antagonist TCP appeared to be very effective in the treatment of casualties who suffered from soman-induced seizures for 30 min: (i)Seizures were arrested within minutes after the TCP injection, confirmed by quantitative electroencephalogram (EEG), after fast Fourier analysis. Three hours after TCP the quantitative EEGs were completely normal in all frequency bands and remained normal during the entire 3-week intoxication period. The power shift to the lower (delta) frequency bands, indicative for neuropathology and found in control animals intoxicated only by soman, was not observed in the soman-TCP group. (ii)The gross neuropathology found in soman control animals within 48 h after soman was prevented in soman-TCP animals and was still absent in 3-week survivors. Instead, ultrastructural changes were observed, indicative of defense mechanisms of the cell against toxic circumstances. (iii)Twenty-four hours after soman, soman-TCP animals were able to perform in the shuttle box and Morris water maze. The beneficial effects of TCP on the performance in these tests during the 3-week intoxication period were very impressive, notwithstanding (minor) deficits in memory and learning. (iv)The increase in excitability after TCP was confirmed by an increase in the acoustic startle response. Taken together, these results confirmed the involvement of NMDA receptors in the maintenance of soman-induced seizures and the development of brain damage. They underline the current hypothesis that cholinergic mechanisms are responsible for eliciting seizure activity after soman and that, most likely, the subsequent recruitment of other excitatory neurotransmitters and loss of inhibitory control are responsible for the maintenance of seizures and the development of subsequent brain damage. PMID- 11920923 TI - Intramuscular diazepam pharmacokinetics in soman-exposed guinea pigs. AB - Intramuscular (i.m.) diazepam is included by the US military as an anticonvulsant in the standard therapeutic regimen for organophosphorus nerve agent intoxication. In this study we investigated the pharmacokinetics of diazepam after i.m. administration while monitoring pharmacodynamic (electroencephalogram, EEG) data in soman-exposed guinea pigs. Prior to experiments the animals were surgically implanted with EEG leads to monitor seizure activity. For the study, animals were administered pyridostigmine (0.026 mg x kg(-1) i.m.) 30 min prior to soman (56 microg x kg(-1), 2 x LD50; subcutaneously, s.c.), which was followed in 1 min by atropine sulfate (2 mg x kg(-1) i.m.) and pralidoxime chloride (25 mg x kg(-1) i.m.). All animals receiving this regimen developed seizure activity. Diazepam (10 mg x kg(-1) i.m.) was administered 5 min after onset of seizure activity. Based on EEG data, animals were categorized as either seizure terminated or not terminated at 30 min after diazepam. Serial blood samples were obtained from each animal. Diazepam (10 mg x kg(-1) i.m.) terminated seizure activity in 52% of the animals within 30 min. The pharmacokinetics were characterized by a one-compartment model with first-order absorption and elimination. The maximum plasma concentrations (Cmax) were 991 and 839 ng x ml( 1) for seizure terminated and not terminated, respectively. Mean plasma concentrations of diazepam were significantly different (P < 0.05) for seizure terminated vs not terminated groups at 30 min. The plasma Cmax in seizure terminated animals in this study is similar to the minimum range of plasma diazepam (200-800 ng x ml(-1)) reported to suppress seizure activity in humans. It has been reported in an earlier study that the minimum effective i.m. dose (0.1 mg x kg(-1)) required to prevent soman-induced convulsions in Rhesus monkeys produces a mean Cmax of 50 ng x ml(-1) for diazepam. The data from our current study suggest that a higher dose (and corresponding Cmax) is necessary to terminate ongoing seizure activity. PMID- 11920924 TI - Prophylaxis against organophosphate poisoning by sustained release of scopolamine and physostigmine. AB - Protection efficacy of continuous prophylactic administration of physostigmine and scopolamine against sarin-induced toxicity was evaluated previously in guinea pigs. The present study in large animals used Beagle dogs, that serve as an animal model with cholinergic sensitivity similar to that of humans. Pretreatment with physostigmine salicylate and scopolamine hydrochloride at dose rates of 2.5 and 1 microg x kg(-1) x h(-1), respectively, was administered via Alzet mini osmotic pumps. At the time of exposure, the physostigmine salicylate concentration in plasma was 0.7 ng x ml(-1) and the scopolamine hydrochloride concentration was ca. 0.2 ng x ml(-1), both of which are levels known to be well tolerated in humans. Whole-blood cholinesterase inhibition was 15-20%. This regimen conferred full protection against 2.5 x LD50 i.v. of sarin. Albeit the high-dose exposure, cholinergic toxicity symptoms were mild with no convulsions. About 11-14 min following poisoning the treated animals started to walk and 15-20 min following exposure full recovery was observed and the dogs behaved normally. With higher dose rates of physostigmine salicylate and scopolamine hydrochloride, at plasma concentrations of 2.1 and 0.6 ng x ml(-1), respectively, treated dogs regained normal posture 6-10 min after exposure. PMID- 11920925 TI - Cardiopulmonary effects of HI-6 treatment in soman intoxication. AB - The cardiopulmonary effects of HI-6, together with atropine and soman, were studied in the rat. HI-6 is an effective antidote in acute poisoning with the nerve agent soman. The therapeutic efficiency of HI-6 is still unclear and cannot be explained entirely by the HI-6 reactivating ability of acetylcholinesterase (AChE). Other non-cholinergic factors must be involved. One possible detoxifying process might be an effect of HI-6 on the blood flow to sensitive organs. The purpose of the present study was to investigate 1) whether soman per se induces changes in regional blood flow and 2) whether the blood flow to different organs is affected when HI-6 (50 mg x kg(-1) i.m.) and atropine (10 mg x kg(-1) i.m.) are given either before or immediately after soman intoxication (90 microg x kg( 1) s.c.). For regional blood flow determinations the microsphere method was used with male Wistar rats weighing 300-400 g. The rats were anaesthetised and breathed spontaneously during the experiment. Three different blood flow measurements were made in the same animal and concomitant physiological parameters such as mean arterial blood pressure and respiratory rate were recorded. The blood AChE activity was followed throughout the experiment. Our results show that when HI-6 is given after intoxication with soman, dramatic changes in blood flow occur with a significant decrease in both respiratory rate and blood AChE activity. If HI-6 is given prior to the intoxication, however, all rats are unaffected and none of the parameters measured are changed. PMID- 11920926 TI - An improved brain slice model of nerve agent-induced seizure activity. AB - A brain slice model was developed to investigate the mechanisms of seizure activity induced by soman and the effectiveness of potential anticonvulsant drugs. Unlike previously reported slice studies with nerve agents, this model contains the entorhinal cortex as well as the hippocampus. This allows the study of the spread of seizure discharges within the limbic system and the development of prolonged, sustained discharges that are rarely seen in the simple hippocampal slice preparation. Soman (1 microM) induced a second population spike in the evoked field potential in the CA1 or CA3 region within 15-20 min. In almost all the slices tested, this developed into spontaneous seizure activity within 30-40 min. As well as interictal bursts, many slices also showed longer periods of high frequency bursting analogous to ictal seizure activity that originated in the entorhinal cortex. This activity appeared similar to that induced by the muscarinic agonist pilocarpine. Both the second population spike and the spontaneous discharges could be blocked by diazepam and by AMPA/kainate antagonists, but not by the NMDA antagonists AP5 and MK-801. This study confirms that the combined hippocampal-entorhinal cortex slice preparation is a suitable model for investigating the origin and propagation of nerve-agent-induced seizures within the limbic system. PMID- 11920927 TI - Soman-induced seizures: limbic activity, oxidative stress and neuroprotective proteins. AB - Soman, a potent acetylcholinesterase inhibitor, induces status epilepticus in rats followed by conspicuous neuropathology, most prominent in piriform cortex and the CA3 region of the hippocampus. Cholinergic seizures originate in striatal nigral pathways and with fast-acting agents (soman) rapidly spread to limbic related areas and finally culminate in a full-blown status epilepticus. This leads to neurochemical changes, some of which may be neuroprotective whereas others may cause brain damage. Pretreatment with lithium sensitizes the brain to cholinergic seizures. Likewise, other agents that increase limbic hyperactivity may sensitize the brain to cholinergic agents. The hyperactivity associated with the seizure state leads to an increase in intracellular calcium, cellular edema and metal delocalization producing an oxidative stress. These changes induce the synthesis of stress-related proteins such as heat shock proteins, metallothioneins and heme oxygenases. We show that soman-induced seizures cause a depletion in tissue glutathione and an increase in tissue 'catalytic' iron, metallothioneins and heme oxygenase-1. The oxidative stress induces the synthesis of stress-related proteins, which are indicators of 'stress' and possibly provide neuroprotection. These findings suggest that delocalization of iron may catalyze Fenton-like reactions, causing progressive cellular damage via free radical products. PMID- 11920928 TI - Effects of selected anticholinergics on acoustic startle response in rats. AB - The present study compared the effects of the anticholinergics aprophen hydrochloride, atropine sulfate, azaprophen hydrochloride, benactyzine hydrochloride, biperiden hydrochloride, diazepam, procyclidine hydrochloride, scopolamine hydrobromide and trihexyphenidyl hydrochloride on acoustic startle response in rats. Peak startle amplitude, latency to peak startle amplitude and prepulse inhibition following 100- and 120-dB tones were recorded 15 min following drug administration in food-restricted rats. Aprophen, atropine, azaprophen, benactyzine, biperiden and scopolamine significantly increased peak startle amplitude and decreased latency to peak startle amplitude following 100 dB pulses. In contrast, only biperiden increased peak startle amplitude following 120-dB pulses, whereas atropine and trihexyphenidyl decreased latency to peak startle amplitude following 120-dB pulses. Benactyzine decreased prepulse inhibition following both 100- and 120-dB pulses, whereas both biperiden and scopolamine decreased prepulse inhibition following 120-dB pulses. Acoustic startle response measures were effective in differentiating the effects of anticholinergic compounds. The comparison of drug effects on the acoustic startle response may be useful in selecting efficacious anticholinergic drug therapies with a minimal range of side-effects. In addition, these data may be useful in down-selecting the number of anticholinergic drugs that need to be tested in comparison studies involving more complex behavioral tests. PMID- 11920929 TI - Carboxylesterase: specificity and spontaneous reactivation of an endogenous scavenger for organophosphorus compounds. AB - The ability of carboxylesterase (CaE) to act as a bioscavenger to provide protection against organophosphorus (OP) compounds has been demonstrated in several animal models. To further evaluate the effectiveness of CaE as a bioscavenger, the specificity and stoichiometry of the detoxication of OP compounds by rat plasma CaE were examined. The specificity of CaE was evaluated by determining the bimolecular rate constants for inhibition (k(i)) of CaE by a variety of OP compounds. CaE exhibited a broad specificity for neutral OP compounds with k(i) > 10(6) M(-1) x min(-1) for paraoxon, sarin, soman, diisopropyl fluorophosphate, and diphenyl p-nitrophenyl phosphinate. CaE exhibited poor reactivity (k(i) < 10(4) M(-1) x min(-1)) with cationic OP compounds, such as echothiophate, VX, and iso-OMPA. The stoichiometry of CaE detoxication of OP compounds was evaluated by determining the rates of enzyme reactivation and ageing of OP-inhibited CaE. CaE exhibited no ageing after inhibition by any of the OP compounds, including soman. However, OP-inhibited CaE did exhibit spontaneous reactivation with reactivation rates that decreased as the size of the OP increased (i.e., VX > sarin > soman). The pH dependence of the spontaneous reactivation of sarin-inhibited CaE suggested that its reactivation was dependent on an amino acid residue with a pK(a) of 6.1, which is probably a histidine that is highly conserved in CaE but not in other esterases. PMID- 11920930 TI - Effects of advanced candidate anticonvulsants under two rodent models of 'counting'. AB - The behavioral effects of a variety of advanced candidate anticonvulsants for organophosphate-induced seizures were evaluated under two rodent 'counting' models. Rats pressed the left of two levers a number of times (a 'run') before pressing the right lever. The targeted performance was a run of 12. The training contingency was a targeted percentile schedule, which provided food if the current run was closer to 12 than two-thirds of the most recent runs. Baseline performance was well controlled by the target, with mean run lengths slightly less than 12. Once this performance was acquired, half the subjects were switched to a procedure providing food following runs of different lengths with a probability yoked to previous percentile schedule performance. The two procedures generate comparable baseline performances, but behavioral disruptions generate reinforcement loss only under the yoked procedure. Atropine, scopolamine, azaprophen, aprophen, trihexyphenidyl, procyclidine, benactyzine, biperiden and diazepam were tested. All produced dose-related decreases in overall run length and response rate. Responding was disrupted more readily under the yoked procedure than under the percentile procedure. Only atropine affected responding at doses below those effective against soman-induced seizures. Of the present candidates, trihexyphenidyl, procyclidine, benactyzine and biperiden appear most promising for further development. PMID- 11920931 TI - Bretazenil, a benzodiazepine receptor partial agonist, as an adjunct in the prophylactic treatment of OP poisoning. AB - Benzodiazepines, mainly diazepam, are commonly used as anticonvulsants in the treatment of organophosphate casualties. Although very effective, diazepam usually is not used in prophylactic treatments because of its adverse effects on task performance and its abuse liability. Benzodiazepine (BZ) partial agonists are unique in that they are able to occupy all the population of a given receptor without eliciting the maximal physiological response. The BZ receptor agonistic occupancy was found to differ among the various physiological responses in the following order: antipanic > anticonvulsion > sedation > muscle relaxation. Thus, partial agonists, by the use of which controlled levels of agonistic activity can be achieved, might serve as effective anticonvulsants, with fewer side-effects. Bretazenil, a partial agonist, was found to counteract metrazol-induced convulsions in rats. At the anticonvulsive doses (125-250 microg x kg(-1), i.p.) bretazenil, in combination with pyridostigmine (100 microg x kg(-1), i.m.) and aprophen (4 mg x kg(-1), i.m.), conferred prophylactic protection against sarin and soman poisoning (protective ratios 2.6 and 2.1, respectively). Relevant doses of bretazenil (50-400 microg x kg(-1), i.p.) also were tested for general behavioural effects in the open field and for its anti-anxiety properties in the plus maze. The incapacitation was much lower compared with diazepam. Bretazenil should be considered as a candidate for incorporating into a prophylactic mixture as a central nervous system protectant, with significant advantages concerning incapacitation. PMID- 11920932 TI - Respiratory hypersensitivity to trimellitic anhydride in Brown Norway rats: a comparison of endpoints. AB - A rat bioassay has been developed to provide an objective approach for the identification and classification of respiratory allergy using trimellitic anhydride (TMA), which is a known respiratory tract irritant and asthmagen. Particular emphasis was placed on the study of route-of-induction-dependent effects and their progression upon inhalation challenge with TMA (approximately 23 mg m(-3) for a duration of 30 min), which included analysis of specific and non-specific airway hyperreactivity and pulmonary inflammation initiated and sustained by immunological processes. Refinement of the bioassay focused on procedures to probe changes occurring upon challenge with TMA or methacholine aerosols using physiological, biochemical and immunological procedures. Following challenge with TMA, the rats sensitized to TMA showed marked changes in peak inspiratory and expiratory air flows and respiratory minute volume. In these animals, a sustained pulmonary inflammation occurred, characterized by specific endpoints determined in bronchoalveolar lavage (lactate dehydrogenase, protein, nitrite, eosinophil peroxidase, myeloperoxidase). When compared with the naive controls, lung weights were increased significantly, as were the weights of lung associated lymph nodes following inhalation induction and auricular lymph nodes following topical induction. The extent of changes observed was equal or more pronounced in animals sensitized epicutaneously (day 0:150 microl vehicle/50% TMA on each flank, day 7; booster administration to the skin of the dorsum of both ears using half the concentration and volume used on day 0) when compared with rats sensitized by 5 x 3 h day(-1) inhalation exposures (low dose: 25 mg TMA m( 3), high dose: 120 mg TMA m(-3)). In summary, the findings support the conclusion that the Brown Norway rat model is suitable for identifying TMA as an agent that causes both an immediate-type change of breathing patterns and a delayed-type sustained pulmonary inflammatory response. However, it remains unresolved whether the marked effects observed in the topically sensitized rats are more related to a route-of-induction or dose-dependent phenomenon. PMID- 11920933 TI - Poisoning in Zimbabwe: a survey of eight major referral hospitals. AB - A retrospective study of the pattern of poisoning cases admitted to eight major urban referral hospitals in Zimbabwe over a 2-year period (1998-1999 inclusive) was conducted to describe the pattern of poisoning at these centres. There were a total of 2764 hospital admissions due to poisoning, involving a total of 2846 toxic agents. Accidental poisoning (AP) and deliberate self-poisoning (DSP) accounted for 48.9% (1352 cases) and 41.3% (1142 cases), respectively. With AP, the highest number of cases (45.9%) occurred in children below the age of 5 years, with half of these due to chemicals, mainly paraffin. In the DSP group, however, more than 60% of all cases occurred in the 16-25-year age group. In addition, twice as many females as males were admitted for DSP compared with an overall male/female ratio of 1 : 1.2. Pesticides (31.4%) and pharmaceuticals (30.4%) were the most common groups of toxic agents responsible for the hospital admissions. Unknown toxins, natural toxins and pesticides showed the highest mortality rates (15.4%, 8.3% and 6.7%, respectively). Compared with the last major survey of poisoning in Zimbabwe, the pattern of poisoning at referral hospitals has changed over the last decade, with an increase in pesticide and pharmaceutical cases and a marked fall in cases of traditional medicine poisoning. Educational and legislative interventions may be required to address these changes. There is the need also to investigate further the high mortality rates associated with traditional medicine poisoning. PMID- 11920935 TI - Decreased serum testosterone levels in rats exposed intraperitoneally to bismuth subnitrate. AB - The consumption of bismuth is increasing and knowledge of the potential teratogenic and reproductive damage of bismuth exposure is fragmentary. In the present study we used autometallography (AMG)--the histochemical silver amplification technique--to trace bismuth in the testis and pituitary glands of Wistar rats injected intraperitoneally with bismuth subnitrate. Large amounts of bismuth AMG grains were concentrated in the lysosomes of Leydig cells, and serum testosterone levels were reduced when compared with controls. No histochemical traces of bismuth were found in the anterior lobe of the pituitary gland. Compared with their corresponding controls, neither follicle-stimulating hormone nor luteinizing hormone were affected. The selective uptake of bismuth in Leydig cells, followed by decreased testosterone levels, emphasizes a potential hazard of bismuth-provoked male reproductive impairment. PMID- 11920934 TI - Influence of water extracts from the surface of two yew (Taxus) species on mites (Tetranychus urticae). AB - Taxus cuspidata and Taxus media var. Hicksii contain paclitaxel, among other taxoids, on the surface of the needles. These compounds were removed by 5-s dipping of the needles in water just below its boiling point at 96 degrees C and at 60 degrees C and 40 degrees C. Taxus cuspidata contained a fourfold higher concentration of paclitaxel than Taxus media var. Hicksii. The extract with the higher concentration of paclitaxel was more harmful to the mites Tetranychus urticae Koch, increasing their mortality 150%, prolonging development by ca. 20% and lowering the average fecundity from 112 in the control to 16.13 after treatment with Taxus cuspidata; also, the net reproductive rate dropped from 70.24 to 6.70, which is more than a tenfold reduction. PMID- 11920936 TI - Determination of the highest no-effect dose (HNED) and of the elimination pattern for cocaine in horses. AB - Cocaine is one of the most widespread illegal stimulants utilized by the human population throughout the world. The aim of this study was to establish the highest no-effect dose (HNED) of cocaine on the spontaneous locomotor activity (SLA) of horses in a behavior chamber, and thereby to determine the maximal acceptable threshold of the urinary drug concentration in horses. Twelve English thoroughbred mares received 0.02, 0.03, 0.04, 0.08 or 0.12 mg kg(-1) cocaine i.v. or saline solution (control). It was noted that doses above 0.04 mg kg(-1) induced a significant increase in SLA (P < 0.05, Tukey's test). No significant increase in SLA was seen in the mares that received 0.03 mg kg(-1), but the animals showed important behavioral changes that did not occur after the 0.02 mg kg(-1) dose. It was concluded that the HNED of cocaine for horses in a behavior chamber is 0.02 mg kg(-1). After injection of this dose in five horses, urine samples were collected at predetermined intervals through vesical catheterization. The concentrations of cocaine, norcocaine, benzoylecgonine and ecgonine methyl ester were quantified by liquid chromatography/electrospray ionization tandem mass spectrometry. Cocaine and norcocaine concentrations remained consistently below the level of detection. Benzoylecgonine reached a mean (+/- SEM) maximum concentration of 531.9 +/- 168.7 ng ml(-1) after 4 h, whereas ecgonine methyl ester peaked 2 h after injection at a concentration of 97.2 +/- 26.5 ng ml(-1). The maximum admissible concentration for cocaine and/or metabolites in the urine of horses is difficult to establish unequivocally because of the substantial individual variation in the drug elimination pattern observed in horses, which can be inferred by the large standard error of the means obtained. PMID- 11920937 TI - Development and validation of a high-performance liquid chromatography method for the determination of cocaine, its metabolites and ketamine. AB - Cocaine abuse is an extensive problem in the USA. During the past decade, ketamine abuse also has emerged as a public health concern and is now considered a controlled substance. The prevalence of the simultaneous use of cocaine and ketamine has been shown to be high. Previous research indicates that ketamine affects the enzymes that metabolize cocaine. In order to investigate this pharmacokinetic interaction, it was necessary to identify and quantitate each compound. The aim of this study is to develop a method of detecting and resolving cocaine, its metabolites and ketamine. A new precise, accurate and sensitive reversed-phase high-performance liquid chromatography method has been developed and validated. This assay employed a phosphate-buffered aqueous mobile phase (pH 6.9) with an organic component consisting of acetonitrile and methanol and a C-18 column as stationary phase at 225 nm wavelength. Minimum detection limits were 5 ng ml(-1) for cocaine and 10 ng ml(-1) for benzoylecgonine, norcocaine and ketamine. Linearity was demonstrated over a broad range of concentration in plasma, with good sensitivity for ketamine, cocaine and cocaine metabolites. PMID- 11920938 TI - Sexually dimorphic alterations of brain cortical dominance in rats prenatally exposed to TCDD. AB - Sexually dimorphic patterns of cortical lateralization are documented extensively in both human and animal brains. Male rats tend to exhibit pronounced right hemisphere dominance compared with females, whereas females typically exhibit more diffuse lateralization patterns and greater left hemisphere bias compared with males. Prenatal TCDD (2,3,7,8 tetrachlorodibenzo-p-dioxin) exposure produces demasculinization of male offspring sexual behavior. In previous studies, we showed a reversal of cortical dominance in rats after prenatal TCDD exposure on gestational day 18 (GD 18). The current study aimed to determine the nature of changes observed in rats exposed to TCDD on GD 8. In addition, locomotor activity was measured in male and female offspring on postnatal day (PND) 30, 60 and 90. Pregnant females were given, via gavage, a single dose of 0, 20, 60 or 180 ng kg( 1) TCDD on GD 8. Cortical depth measurements were taken in selected brain regions in offspring 3 months old that had been exposed to the 180 ng kg(-1) dose. Areas 2, 3, 17, 18a and 39 at bregmas -1.8, -3.8 and -5.8 were analyzed by quantifying digitized, enhanced images produced by a photomicroscope fitted with a special color camera. In both male and female offspring, cortical thicknesses in control brains exceeded those of exposed brains. In several brain areas of male offspring exposed to TCDD, right hemispheric dominance reversed to left hemispheric dominance. Female offspring brains showed a contrary move towards right hemisphere dominance. Motor activity in juvenile and mature animals did not differ among dose groups. These data demonstrate that prenatal exposure to TCDD reduces cortical thickness and alters the normal pattern of cortical asymmetry, a finding consistent with the sexually dimorphic behavioral effects induced by this agent. PMID- 11920939 TI - Sulfur mustard-stimulated protease: a target for antivesicant drugs. AB - One of the mechanisms of the skin blistering effect (vesication) of sulfur mustard (bis-(2-chloroethyl)sulfide, HD) is believed to be via the stimulation of specific protease(s) at the dermal-epidermal junction. Cultured normal human epidermal keratinocytes (NHEK) were used as a model to study and characterize protease stimulated by the mustards 2-chloroethyl ethyl sulfide (CEES), 2-chloro N-(2-chloroethyl)-N-methylethanamine hydrochloride (nitrogen mustard, HN(2)) and HD. The results obtained using a chromozym (TRY) peptide substrate protease assay revealed the optimum mustard concentrations and time for protease stimulation to be about 200 microM (CEES), 100 microM (HN(2)) and 100 microM (HD) and 16 h. The mustard-stimulated protease was membrane bound and was inhibited by adding a Ca(2+) chelator (either 2 mM EGTA (ethylene glycol-bis(amino ethyl ether) N,N,N',N' tetraacetic acid) or 50 microM BAPTA AM (1,2-bis(z-aminophenoxy)ethane N,N,N',N'-tetraacetic acid, tetraacetoxy methyl ester) alone or in combination), a serine protease inhibitor diisopropyl fluoro-phosphate (DFP, 1 mM), or a protein synthesis inhibitor cycloheximide (35 microM) in the extracellular medium. These results suggest that mustard toxicity may involve the stimulation of trypsin/chymotrypsin-like serine protease, dependent on Ca(2+) and new protein synthesis. Protein purification by gel exclusion and hydrophobic chromatography produced a 70-80 kDa protease, which had an amino acid sequence homologous with a mammalian-type bacterial serine endopeptidase. Based on this information, research is in progress to identify the protease stimulated by HD in NHEK and to determine whether its inhibitors are useful as prospective antivesicant drugs. PMID- 11920940 TI - Separation of cephalosporins on thin silica gel layers impregnated with transition metal ions and by reversed-phase TLC. AB - Cephalosporin antibiotics were separated on thin layer plates impregnated with transition metal ions, Mn(2+), Fe(2+), Co(2+), Ni(2+) and Cu(2+), using different concentrations. Various solvent systems were developed for the study and were used for separation of these analytes. Impregnation was observed to have an effect on hR(F) values, removed tailing of analytes and improved the resolution. The results have been discussed for each metal ion and compared, and the best conditions of separation have been identified. Activation time of thin layer plates impregnated with 0.1% FeSO(4) was found to affect both hR(F) values and resolution of cephalosporins. New solvent systems are reported for both normal phase and reversed-phase TLC. PMID- 11920941 TI - High performance liquid chromatographic-mass spectrometric assay for the quantitation of BMS-204352 in dog K(3)EDTA plasma. AB - A high performance liquid chromatographic-mass spectrometric (LC/MS) assay was developed and validated for the determination of BMS-204352 in dog K(3)EDTA plasma. A 0.5 mL aliquot of control plasma was spiked with BMS-204352 and internal standard (IS) and buffered with 1 mL of 5 mM ammonium acetate. The mixture was then extracted with 3 mL of toluene. After separation and evaporation of the organic phase to dryness using nitrogen at 40 degrees C, the residue was reconstituted in the mobile phase and 25 microL of the sample were injected onto a Hypersil C(18) column (2 x 50 mm; 3 microm) at a flow rate of 0.5 mL/min. The mobile phase was consisted of two solvent mixtures (A and B). Solvent A was composed of 5 mM ammonium acetate and 0.1% triethylamine in 75:25 v/v water:methanol, pH adjusted to 5.5 with glacial acetic acid, and solvent B was 5 mM ammonium acetate in methanol. A linear gradient system was used to elute the analytes. The mass spectrometer was programmed to admit the de-protonated molecules at m/z 352.7 (IS) and m/z 357.9 (BMS-204352). Standard curves of BMS 204352 were linear (r(2) > or = 0.998) over the concentration range of 0.5-1000 ng/mL. The mean predicted quality control (QC) concentrations deviated less than 5.1% from the corresponding nominal values (ie 4, 80, 400 and 2000 ng/mL); the within- and between-assay precision of the assay were within 5.5% relative standard deviation. Stability of BMS-204352 was confirmed after at least three freeze/thaw cycles and BMS-204532 was stable in dog plasma when stored frozen at or below -20 degrees C for at least 16 weeks in spiked QC samples and for at least 4 1/2 weeks for in vivo study samples. BMS-204352 and IS were stable in the injection solvent at room temperature for at least 24 h. The assay was applied to delineate the pharmacokinetic disposition of BMS-204352 in dogs following a single intravenous dose administration. In conclusion, the assay is accurate, precise, specific, sensitive and reproducible for the pharmacokinetic analysis of BMS-204532 in dog plasma. PMID- 11920942 TI - An automated fluorescence protein sequencer using 7-methylthio-4-(2,1,3 benzoxadiazolyl) isothiocyanate (MTBD-NCS) as an Edman reagent. AB - An automated fluorescence protein sequencer using 7-methylthio-5-(2,1,3 benzoxadiazolyl) isithiocyanate (MTBD-NCS), a fluorescent Edman reagent, is developed by the modification of a commercial protein sequencer. The generated MTBD-thiohydantoin amino acids fluoresced strongly, whereas the by-products such as MTBD-thiocarbamoyl amino acids and MTBD-carbamoly amino acids did not fluoresce. A few interfering peaks were observed in the chromatogram and amino acid sequence was easily determined. The coupling and cyclization/cleavage reaction conditions and extraction conditions of generated MTBD-thiazolinone amino acids were optimized using an autonalyzer. Finally, the sequence of a synthetic peptide (25 pmol), leucine-enkephalin-Thr-amide, was determined and up to six residues were successively analyzed. PMID- 11920943 TI - Simultaneous determination of ondansetron and tropisetron in human plasma using HPLC with UV detection. AB - A rapid and sensitive HPLC method for the simultaneous quantitation of ondansetron and tropisetron, two serotonin (5-HT) receptor antagonists frequently used in treatment and prevention of nausea and emesis, is described. The procedure involves liquid-liquid extraction of human plasma with dichloromethane coupled with reversed-phase HPLC and UV detection. The lower limits of quantification (LOQ) were 0.62 ng/mL for ondansetron and 1.25 ng/mL or tropisetron. Intra- and inter-assay coefficients of variation ranged from 1.5 to 7.5% and 5.3 to 13.7%, respectively. The sensitivity and precision were sufficient for determination of plasma concentrations after therapeutic administration of both drugs and the method can be used for the estimation of pharmacokinetic parameters. PMID- 11920944 TI - Chiral liquid chromatography tandem mass spectrometry in the determination of the configuration of glyceric acid in urine of patients with D-glyceric and L glyceric acidurias. AB - Glyceric acid is a highly polar chiral carboxylic acid that is usually not detected during routine organic acid analysis. Increased excretion is observed in two phenotypically distinct and rare inherited metabolic diseases, D-glyceric aciduria, and L-glyceric aciduria (also known as primary hyperoxaluria type 2). The determination of the exact configuration of the excreted glyceric acid is necessary for the accurate diagnosis of D-glyceric aciduria and for the differentiation between type 1 and type 2 primary hyperoxaluria. The separation of the two stereoisomers was achieved using a narrow-bore ristocetin A glycopeptide antibiotic silica gel bonded column. Triethylamine acetate at pH 4.1 with 10% methanol was used as mobile phase. The column was directly interfaced to a triple quadrupole tandem mass spectrometer and the electrospray ion source was operated in the negative ion mode. Three parent-to-daughter transitions were employed to specifically detect eluting glyceric enantiomers from essentially untreated urine samples. The two forms of glyceric acid were satisfactorily separated at 3.6 and 4.5 min. Application of the method led to the confirmation of three cases of D-glyceric aciduria from three different families. Two other cases are suspected to be L-glyceric aciduria but further confirmation is needed. The method allowed the detection of the glyceric acid stereoisomers in control urine where it was found without exception that L-glyceric was the predominate metabolite. PMID- 11920945 TI - Detection of oxidative stress. Interest of GC-MS for malondialdehyde and formaldehyde monitoring. AB - Ischemia-reperfusion syndrome is a condition where the role of oxygen free radicals is important. Malondialdehyde (MDA) and formaldehyde (FA), products of lipid peroxidation, are the presumptive markers for the development of oxidative stress in tissues and plasmas. A GC-MS method for the determination of MDA and FA in rat brain extract is described. Rat brains were homogenized with deionized water. The homogenates were derivatized with 2,4-dinitrophenylhydrazone (DNPH) to obtain hydrazines derivatives of MDA and FA. The hydrazine derivatives were analyzed by GC-MS and quantitation was by single ion monitoring (SIM). The retention times of FA and MDA were, respectively, 13.75 and 14.20 min, and for SIM quantitation, ion at m/z 210 for FA, and m/z 158 for MDA were used. The results showed that it is possible to estimate the products of lipid peroxidation in brain and to monitor the oxidative stress developed during the ischemia reperfusion syndrome compared to the normal values. PMID- 11920946 TI - High-performance liquid chromatographic method with ultraviolet detection for the determination of dapsone and its hydroxylated metabolite in human plasma. AB - A validated high-performance liquid chromatographic method with ultraviolet detection for the quantitative determination of dapsone (4,4'-diaminodifenyl sulfone, DDS) and a metabolite, hydroxylaminodapsone (4-amino-4 hydroxylaminodiphenyl sulfone, DDS-NOH), in human plasma is described. Human plasma was deproteinized with acetone and the clear supernatant solution after centrifugation was evaporated to dryness under a gentle stream of nitrogen at 70 degrees C. The residue was dissolved in a mixture of HPLC eluent and acetone (18:5 v/v) and an aliquot of this solution (50 microL) was injected onto the HPLC column. Dapsone, hydroxylaminodapsone and diazoxide as internal standard, were separated within 10 min by isocratic elution with water:acetonitrile:glacial acetic acid:triethylamine (80:20:1.0:0.5 by volume) as eluent. Detection was by ultraviolet at the wavelength of 295 nm. The within-day repeatability coefficients of variation were 3-5% for dapsone (0.301-20.0 mg/L, n = 5) and 3-5% for hydroxylaminodapsone (0.0948-6.32 mg/L, n = 5), whereas the between-day repeatability coefficients of variation were 3-8% (0.301-20.0 mg/L, n = 5) for dapsone and 4-10% for hydroxylaminodapsone (0.0948-6.32 mg/L, n = 5). The mean recoveries -were 92-107% (0.301-20.0 mg/L, n = 2), 80-82% (0.0948-6.32 mg/L, n = 2) and 88% (0.0200 mg/mL, n = 5), for dapsone, hydroxylaminodapsone and diazoxide, respectively. The average correlation coefficient of the calibration curve was 0.99988 (n = 5) for dapsone at a concentration range of 0.301-20.0 mg/L, whereas the average correlation coefficient of the hydroxylaminodapsone calibration curve was 0.99981 (n = 5) at a concentration range of 0.0948-6.32 mg/L. The limits of detection were 0.00200 and 0.0470 mg/L for dapsone and hydroxylaminodapsone, respectively. The method is suitable for drug level monitoring and for pharmacokinetic studies. PMID- 11920947 TI - An analytical method for irinotecan (CPT-11) and its metabolites using a high performance liquid chromatography: parallel detection with fluorescence and mass spectrometry. AB - Irinotecan or 7-ethyl-10-[4-(1-piperidino)-1-piperidino]carbonyloxycamptothecin (CPT-11) is an anticancer pro-drug used in the treatment of many types of cancer. We describe here the validation of an analytical method for CPT-11 and its metabolites, including an active metabolite, 7-ethyl-10-hydroxycamptothecin (SN 38), its glucuronidated form, SN-38G, and several cytochrome P450 3A-mediated products such as 7-ethyl-10-[4-N-(5-aminopentanoic acid)-1 piperidino]carbonyloxycamptothecin (APC) using a high-performance liquid chromatography connected to parallel fluorescence and mass spectrometry detection systems. This method is characterized as follows: (1) simple extraction of the analytes from biomaterials with perchloric acid/methanol; (2) sensitive quantitation of major metabolites (SN-38G, SN-38 and APC) with a fluorescence detector (FLD), where the limits of quantitation by FLD were 2.5 ng/mL for SN-38G and APC, 5 ng/mL for CPT-11 and 1 ng/mL for SN-38, respectively; (3) parallel selective monitoring of the metabolites including minor metabolites with a mass selected detector (MSD). There was no observed interference by other drugs expected to be co-administered. This method showed its usefulness by identifying a novel metabolite produced in human hepatic microsomes. The results indicate that this combination of FLD and MSD enables a highly selective analysis of CPT 11 and its metabolites, and is useful for studies both in vivo and in vitro. PMID- 11920948 TI - Disposition of triazolam in the rat by brain microdialysis and semi-micro column high-performance liquid chromatography with UV absorbance detection. AB - A semi-micro column high-performance liquid chromatography with ultraviolet detection for the determination of triazolam is described. The method was applied to determine plasma and brain microdialysate concentrations of triazolam after single intravenous bolus of 2.5 mg/kg to rat. The separation was achieved on a 250 x 1.5 mm i.d. C(18) column and the column effluent was monitored at 222 nm. The detection limits at a signal-to-noise ratio of 3 obtained using spiked plasma and artificial cerebrospinal fluid were 2.1 and 0.7 ng/mL, respectively. The intra- and inter-day reproducibility of the present method were satisfactory with the highest relative standard deviation of 9.1 (n > or = 5). The present method was successfully applied to study the disposition of triazolam in rat (n = 5) by analyzing plasma and brain microdialysate samples. PMID- 11920949 TI - Simultaneous analysis of oxidized and reduced glutathione in cell extracts by capillary zone electrophoresis. AB - Glutathione (GSH) and glutathione disulfide (GSSG) levels in cells constitute a thiol redox system. They can be used as an indicator of oxidative stress of the cell. In this study, a capillary zone electrophoresis (CZE) method is described that enables quantitation of GSH and GSSG from cellular extracts. The CZE buffer used was 20 mM ammonium acetate containing 5% (v/v) acetic acid at pH 3.1 in conjunction with a polybrene coated capillary operated in reverse polarity mode. Effects of different acids used to prepare cell samples were investigated on CZE performance. The acids include meta phosphoric acid (MPA), trichloroacetic acid (TCA), phosphoric acid (PA) and sulfosalicylic acid (SSA) and are used to stabilize GSH and GSSG before performing CZE analysis. The method features a limit of detection of 4 microM and a limit of quantitation of 12 microM for both GSSG and GSH and recoveries of 94% for GSH and 100% for GSSG. Quantitative analysis of GSSG and GSH in HaCaT cell extracts (5% SSA, w/v) was performed with this method and changes in the ratio of GSH to GSSG in N-ethylmaleimide treated cell sample was observed by comparing with control cell samples. PMID- 11920950 TI - Analysis of menthol in three traditional Chinese medicinal herbs and their compound formulation by GC-MS. AB - A GC-MS method was developed for identification and determination of menthol in three traditional Chinese medicinal herbs and their compound formulation, a granule for treating colds. Volatile oil was simultaneously distilled and extracted into ethyl ether in a unique glass extractor. The separation was performed on an HP-5 MS column. The standard addition method was used for quantitative determination of menthol content in herbal materials and in the granule. A component in the samples was chosen as the internal standard. The contents were calculated with the ratio of peak area percentage. Menthol was identified as the main component in Mentha haplocalyx Briq., and also existed in spikes of Schizonepeta tenuifolia Briq., Folium perilla frutescens (L.) Britt. and granules. The quantitative calibration range was 0.21-10.5 mg/mL. Good precision was demonstrated by an RSD < 4.0%. The mean content of menthol in Mentha haplocalyx Briq., spikes of Schizonepeta tenuifolia Briq., Folium perilla frutescens (L.) Britt. and granules was 121, 0.234, 1.03 and 1.84 mg/kg respectively. PMID- 11920951 TI - Identification of kappa and lambda chains of the major immunoglobulin G subclasses by capillary zone electrophoresis. AB - Immunoglobulins are present in most tissues and plasma and play crucial role in immune system. Alteration of the levels of the immunoglobulin G (IgG) subclasses (IgG1, IgG2, IgG3 and IgG4) is an indication of a disturbed immunological response. The aim of the present study was the development of a capillary electrophoresis (CE) method for the analysis of IgG subclasses in respect to their variable kappa and lambda chains. Various analytical conditions and CE modes, including capillary zone electrophoresis (CZE), capillary isoelectric focusing (CIEF) and micellar electrokinetic capillary chromatography (MECC) have been thoroughly studied. CZE was found to be the most convenient way to separate IgG subclasses. Three of the human IgG subclasses were resolved using uncoated fused-silica and 50 mM phosphate, pH = 9.3, as operating buffer at 20 kV and detection at 214 nm. IgG1kappa was completely separated from IgG2kappa and IgG3kappa, whereas IgG2kappa co-migrated with IgG4kappa, which is the minor IgG subclass. Under the same conditions IgG4lambda was completely separated from IgG1lambda, IgG2lambda and IgG3lambda, enabling the identification of the various lambda chains. The developed CE method is rapid and can be applied to the identification of the major immunoglobulin G subclasses in respect to their variable kappa and lambda chains. PMID- 11920952 TI - Ecto-diadenosine polyphosphates hydrolase activity on human prostasomes. AB - BACKGROUND: Ecto-diadenosine polyphosphates are ubiquitous compounds with several physiological roles. Ecto-diadenosine polyphosphates hydrolase control their actions by degrading and terminating their signaling. The present work deals with the identification and partial characterization of ecto-diadenosine polyphosphates hydrolase on human prostasomes. METHODS: Reverse-phase and paired ion HPLC techniques have been used. RESULTS: Prostasomes have an ecto-diadenosine polyphosphates hydrolase that leads to the degradation of several diadenosine compounds. Kinetic parameters of the enzyme show that diadenosine tetraphosphate is the preferred substrate that is further metabolized by the prostasome-ecto nucleotidases to adenosine. The ecto-enzyme is bound to the prostasome-membranes through a GPI-anchor and is activated by physiological concentration of Ca+2, Mg+2, and Mn+2. Its optimum pH is also in the slightly alkaline physiological range. Human spermatozoa do not possess this hydrolytic activity, but they can acquire it after fusion with prostasomes. CONCLUSIONS: The existence of an enzyme capable of degrading diadenosine compounds and can be transferred to human spermatozoa suggests new physiological implications for the role of prostasomes in fertilization. PMID- 11920953 TI - Osteoblasts produce soluble factors that induce a gene expression pattern in non metastatic prostate cancer cells, similar to that found in bone metastatic prostate cancer cells. AB - BACKGROUND: Progressive prostate cancer typically metastasizes to bone where prostate cancer cells gain an osteoblast-like phenotype and induce osteoblastic metastases through unknown mechanisms. To investigate the biology of prostate cancer skeletal metastases, we compared gene expression between the non metastatic LNCaP cell line and its derivative cell line C4-2B that metastasizes to bone. METHODS: Total RNA from LNCaP and C4-2B cell lines was isolated and used to probe membrane-based gene arrays (Comparison 1). Additionally, LNCaP cells were incubated in the absence or presence of conditioned media (CM) from a human osteoblast-like cell line (HOBIT) and total RNA from these cells was used to probe gene arrays (Comparison 2). Differential expression of genes was confirmed by RT-PCR. RESULTS: Of the 1,176 genes screened, 35 were differentially expressed between LNCaP and C4-2B cells (Comparison 1). HOBIT-CM induced differential expression of 30 genes in LNCaP cells (Comparison 2). Interestingly, 19 genes that were differentially expressed in C4-2B vs. LNCaP also displayed a similar expression pattern in LNCaPs grown in HOBIT-CM. These genes are primarily involved in motility, metabolism, signal transduction, tumorigenesis, and apoptosis. CONCLUSIONS: These results suggest that osteoblasts produce soluble factors that contribute to the progression of prostate cancer skeletal metastases, including their transition to an osteoblast-like phenotype. Additionally, these data provide targets to explore for further investigations towards defining the biology of skeletal metastases. PMID- 11920954 TI - Ventral prostate predominant l, a novel mouse gene expressed exclusively in the prostate. AB - BACKGROUND: Despite the region-specific nature of human prostate disease, there is a paucity of information regarding the molecular basis of prostate regionalization and patterning. To elucidate genetic mechanisms that underlie prostate growth and development, we investigated differential gene expression in mouse prostate lobes. METHODS: mRNA differential display analysis was used to identify differentially expressed genes during development of ventral, anterior, and dorsolateral prostate lobes. Differential gene expression was confirmed by Northern blot analysis and RT-PCR. RESULTS: A novel gene, Ventral prostate predominant1 (Vpp1) was identified. Vpp1 mRNA was evident in all lobes but accumulated predominantly in the ventral prostate, and was detected on postnatal day 7 through adulthood exclusively in the prostate gland. The steady-state level of Vpp1 mRNA decreased markedly in response to castration, suggesting androgen regulation of Vpp1 expression. Analysis of TRAMP tumors demonstrated a dramatic decrease in the level of Vpp1 mRNA. CONCLUSIONS: The spatial distribution and early postnatal onset of Vpp1 expression is consistent with a role for this gene in prostate regionalization. The absolute prostate specificity of Vpp1 expression may allow this gene to serve as a paradigm to study the molecular basis of gene expression that is restricted exclusively to the prostate gland. PMID- 11920955 TI - Vitamin D receptor gene polymorphism and prostate cancer risk. AB - BACKGROUND: 1,25-dihydroxyvitamin D, the active form of vitamin D, exerts antiproliferative effect on prostatic cells, mediated through the vitamin D receptor. In a case-control study, we examined whether the vitamin D receptor (VDR) gene polymorphism in exon 9 could affect prostate cancer susceptibility. METHODS: One hundred ninety newly diagnosed prostate cancer patients and 190 age matched men with benign prostatic hyperplasia (BPH), in whom the presence of prostate cancer was excluded clinically or histologically, were recruited for this study. The VDR TaqI polymorphism was investigated by polymerase chain reaction (PCR) following restriction fragment length polymorphism using DNA from lymphocytes. Depending on the presence or absence of the TaqI restriction site at the third position of codon 352, patients were classified as TT, Tt, or tt. RESULTS: The frequency of the tt genotype was not significantly different between prostate cancer patients (18%) and controls (12%; P = 0.07). The odds ratio (OR), calculated relative to individuals with the TT genotype was 1.76 (95% confidence limit (CL) = 0.90-3.45). After stratification for Gleason score and prostate specific antigen levels in a case-case comparison (n = 190), no significant associations with the VDR genotypes were detectable either. CONCLUSIONS: In this case-control study of Austrian Caucasians, no statistically significant association of the VDR TaqI polymorphism and prostate cancer risk was found. PMID- 11920956 TI - Identification of single nucleotide polymorphisms in the human kallikrein 10 (KLK10) gene and their association with prostate, breast, testicular, and ovarian cancers. AB - BACKGROUND: The KLK10 gene (also known as the normal epithelial cell-specific 1 gene) is a member of the expanded human kallikrein gene family. Recently, it has been reported that KLK10 is a tumor suppressor gene and that its expression is downregulated in various forms of cancer and cancer cell lines. KLK10 is also upregulated in ovarian cancer. We thus hypothesized that the KLK10 gene may be a target for mutations in various cancers. METHODS: We sequenced the five coding exons of the KLK10 gene using genomic DNA from various tumors, normal tissues, and blood, by PCR amplification and automated sequencing. RESULTS: In none of the tumor-derived DNAs, we identified somatic mutations that could inactivate this gene. However, we identified a prevalent germline single nucleotide variation at codon 50 (exon 3) of this gene [GCC (alanine) to TCC (serine)]. The GCC genotype was less prevalent in prostatic cancer patients in comparison to control subjects (P = 0.027) but no differences were seen with testicular, ovarian, and breast cancer. We also identified four genetic variations in exon 4, at codons106 [GGC (glycine) to GGA (glycine)], codon 112 [ACG (threonine) to ACC (threonine)], codon 141 [CTA (leucine) to CTG (leucine)], and at codon 149 [CCG (proline) to CTG (leucine)]. None of these variations was significantly different between normal subjects and cancer groups. CONCLUSIONS: We found no evidence for somatic mutations of the KLK10 gene in cancers of the prostate, breast, ovary, and testis. The single nucleotide variation at codon 50 appears to be associated with prostate cancer risk. PMID- 11920957 TI - Anoikis is regulated by BCL-2-independent pathways in human prostate carcinoma cells. AB - BACKGROUND: Loss of contact with the extracellular matrix (ECM) triggers a specialized form of apoptosis known as "anoikis" in normal epithelial cells. Dependence on adhesion to ECM is often lost in transformed cells, and the degree of anchorage independence may vary in non-metastatic and metastatic cancer cells. BCL-2 oncoprotein overexpression correlates with the progression and metastases of prostate cancer. Materials and Methods We studied anoikis in suspension cultures of PC-3 and LNCaP prostate carcinoma cells selected for enhanced metastatic potential in vivo and in PC-3 and LNCaP cells stably transfected with BCL-2. Apoptosis-associated DNA fragmentation was measured by agarose gel electrophoresis and propidium iodide staining and flow cytometry. Expression of BCL-2 family polypeptides was determined by immunoblotting. RESULTS: Non metastatic PC-3P cells were significantly more sensitive to anoikis than the metastatic PC-3 variants (PC-3M, PC-3M-PRO-4, and PC-3M-LN-4), but anoikis resistance did not correlate with metastatic potential in LNCaP-derived cell lines. Expression of BCL-2 was higher in metastatic PC-3 and LNCaP subclones compared to isogenic non-metastatic cells, but these levels were not affected by anoikis. Enforced overexpression of BCL-2 did not protect either PC-3P or LNCaP PRO-5 cells from anoikis, even though it rendered them resistant to thapsigargin and inhibited cytochrome c release. Strikingly, cells that died of anoikis maintained their pretreatment levels of BCL-2, whereas the cells that survived anoikis expressed much lower levels of the protein. CONCLUSIONS: Sensitivity to anoikis is regulated by BCL-2 independent mechanisms in LNCaP and PC-3 prostate cancer cells. PMID- 11920959 TI - Complex functions of mutant p53 alleles from human prostate cancer. AB - BACKGROUND: Few studies have used multiple assays to examine the functionality of mutant p53 in prostate cancer (CaP). We employed seven functional assays to study 16 representative mutant p53 alleles, six from localized and ten from metastatic CaP. METHODS: Yeast assays were employed to determine loss of function (LOF), partial function (PF), and dominant-negative status. Assays using p53-null Saos2 cells were used to determine whether mammalian cells transfected with mutant p53 could up-regulate the MDR-1 or PCNA promoters, alter IL-6 expression or confer the ability to grow in soft agar. As a further test of gain of function (GOF), p53-null PC3 cells stably transfected with these mutant p53 alleles were examined for cell cycle distributions. RESULTS: All 16 mutant p53 alleles demonstrated either total or partial LOF. All but one allele also had at least one gain of function; however, the pattern of GOF was different for each mutant allele. Alleles derived from both localized and metastatic CaP had similar GOF characteristics; however, only alleles from metastatic disease had significantly increased S-phase fractions. CONCLUSIONS: Different mutant p53 alleles from CaP had different, complex functional profiles. The lack of predictable patterns for these alleles suggest that each mutation may uniquely affect p53 function. PMID- 11920958 TI - Lysophosphatidic acid stimulates proliferation of cultured smooth muscle cells from human BPH tissue: sildenafil and papaverin generate inhibition. AB - BACKGROUND: The endogenous substance lysophosphatidic acid (LPA) has been found to generate proliferation of cultured smooth muscle cells (SMC). Therefore, the effect of LPA on human benign prostate hyperplasia (BPH) could be of interest. METHODS: The proliferative effect of LPA on cultured human prostatic SMC from specimens obtained at trans-urethral resection of the prostate (TURP) because of BPH, was analyzed by [3H]-thymidine and [35S]-methionine incorporation. In addition, LPA stimulated BPH SMC were treated with papaverin, forskolin, sildenafil or zaprinast, well known to increase the intracellular level of cAMP or cGMP. RESULTS: LPA produced a dose-dependent increase in BPH SMC, both regarding DNA- and protein-synthesis with EC50 values of 3 and 10 microM, respectively. Furthermore, both papaverin, a general phosphodiesterase inhibitor regarding cAMP hydrolyzes, and forskolin, an adenylyl cyclase stimulating agent, inhibited the LPA-stimulated DNA replication in a dose dependent manner with IC50 = 2.5, and 0.35 microM, respectively. cGMP increasing agents, such as the NO donors SIN-1 and SNAP, produced a weak anti-proliferative response. However, both phosphodiesterase 5 inhibitors sildenafil (Viagra) and zaprinast efficiently blocked DNA replication. In addition, when the protein synthesis was examined, we found that the LPA response was significantly inhibited by forskolin and papaverin. CONCLUSIONS: The major conclusion of this investigation is that the endogenous serum component LPA, is able to promote human BPH SMC growth. In addition, our study indicates that cyclic nucleotides can inhibit this effect. Future clinical studies will be needed to determine if different specific phosphodiesterase inhibitors per se or in combination could represent a new therapeutic possibility for the treatment of BPH. PMID- 11920960 TI - Use of a prevention index to identify industries at high risk for work-related musculoskeletal disorders of the neck, back, and upper extremity in Washington state, 1990-1998. AB - BACKGROUND: The prevention of work-related musculoskeletal disorders such as carpal tunnel syndrome and low back disorders has been a focus of international prevention efforts including regulation. This study examines workers compensation claims in Washington State to provide baseline data from which to assess the need and the effects of prevention activities. METHODS: Washington State Fund workers compensation claims for general and selected specific hand/wrist, elbow, shoulder, and back disorders in 1990-1998 as well as general self-insured compensable (four or more lost workdays) claims data were examined. Payroll hours were used to calculate claims incidence rates per 10,000 full-time equivalent employees (FTEs). We created a prevention index (PI) to rank industries by averaging the ranks of their number of claims and their claims incidence rate. The focus was on non-traumatic soft tissue musculoskeletal disorders (NTST-MSDs). RESULTS: Between 1990-1998, there were 392,925 State Fund accepted claims for NTST-MSDs of the neck, back, and upper extremity resulting in $2.6 billion in direct costs and 20.5 million lost workdays. The average claims incidence rate (CIR) was 355 NTST-MSDs per 10,000 FTEs. The NTST-MSD CIR decreased significantly less than that for all other claims (P = 0.05) but the CIR for upper extremity NTST-MSDs did not significantly decrease over the study period. There were no significant changes in the CIRs for sciatica (4.9 per 10,000 FTEs) and rotator cuff syndrome (15.3 per 10,000 FTEs), whereas the CIR for epicondylitis (10.6 per 10,000 FTEs) increased and for carpal tunnel syndrome (24.5 per 10,000 FTEs) decreased significantly over the study period. Based on the prevention index, the top five industries for combined State Fund and Self-Insured Compensable NTST MSDs were Trucking and Courier Services (SIC 421), Nursing Homes (SIC 805), Masonry (SIC 174), Air Transportation (SIC 451), and Residential Construction (SIC 152). Using Washington Industrial Classes (WIC), temporary workers in assembly and administrative services were also high on the prevention index. CONCLUSIONS: NTST-MSDs continue to be a large and costly problem in Washington State. While the incidence rates for some NTST-MSDs are decreasing, the overall rate is not decreasing as fast as the rate for all other claims. In some cases, the rate is stable (sciatica, rotator cuff syndrome) or increasing (epicondylitis). Heavy manual handling and repetitive work characterize the industries with the highest risk. PMID- 11920961 TI - Musculoskeletal problems of the neck, shoulder, and back and functional consequences in nurses. AB - BACKGROUND: Though musculoskeletal disorders (MSDs) are highly prevalent among registered nurses (RNs), little is known about functional consequences of MSDs in nurses. METHODS: Data on neck, shoulder, and back MSD problems were analyzed in 1,163 working nurses (response rate = 74%). Cases had relevant symptoms lasting at least 1 week or occurring at least monthly in the past year, with at least moderate pain intensity, on average. MSD problems with a frequency, duration, or pain intensity below the level needed to meet the case definition were defined as MSD symptoms. Those who did not meet symptom or case criteria at any body site were defined as asymptomatic. Odds of consequences (e.g., saw a doctor/provider, missed work, reduced/modified work, non-work activities, or recreation, medication use, inadequate sleep) were estimated for cases versus those with symptoms. RESULTS: We found 45.8, 35.1, and 47.0% of nurses had neck, shoulder, or back MSD problems (either at the case or symptom level), respectively, within the past year. Cases were far more likely to have seen a provider versus those with symptoms (adjusted odds ratio, aOR Neck: 4.33, 95% CI: 2.85-6.56; aOR Shoulder: 4.83, 95% CI: 3.00-7.77; aOR Back: 3.69, 95% CI: 2.47-5.49). Cases also were more likely to experience all other functional consequences. CONCLUSIONS: MSD consequences are substantial and suggest opportunities for intervention. Future research will examine the impact of work organization and physical demands on MSDs. PMID- 11920962 TI - Lead exposure in Mexican radiator repair workers. AB - BACKGROUND: Lead exposure was investigated among 73 Mexican radiator repair workers (RRWs), 12 members of their family (4 children and 8 wives), and 36 working controls. RRWs were employed at 4 radiator repair shops in Mexico City and 27 shops in Cuernavaca and surrounding areas. METHODS: Exposure was assessed directly through the use of personal air sampling and hand wipe samples. In addition, industrial hygiene inspections were performed and detailed questionnaires were administered. Blood lead levels were measured by graphite furnace atomic absorption spectroscopy (AAS). RESULTS: The mean (SD) values for blood lead of the RRWs, 35.5 (13.5) microg/dl, was significantly greater than the same values for the working controls, 13.6 (8.7) microg/dl; P < 001. After excluding a single outlier (247 microg/m(3)), air lead levels ranged from 0 to 99 microg/m(3) with a mean (SD) value of 19 (23) microg/m(3) (median = 7.9 microg/m(3)). In a final multivariate regression model of elevated blood lead levels, the strongest predictors were smoking (vs. non-smoking), the number of radiators repaired per day on average, and the use (vs. non-use) of a uniform while at work, which were associated with blood lead elevations of 11.4 microg/dl, 1.95 microg/dl/radiator/day, and 16.4 microg/dl, respectively (all P <.05). Uniform use was probably a risk factor because they were not laundered regularly and consequently served as reservoir of contamination on which RRWs frequently wiped their hands. CONCLUSIONS: Lead exposure is a significant problem of radiator repair work, a small industry that is abundant in Mexico and other developing countries. PMID- 11920964 TI - Pesticide use and pesticide-related symptoms among black farmers in the Agricultural Health Study. AB - BACKGROUND: Health effects of pesticides have not been well studied in black farmers. We describe agricultural practices and pesticide-related symptoms in North Carolina black and white farmers participating in the Agricultural Health Study. METHODS: Self-administered questionnaires were completed by 891 black and 11,909 white farmers licensed to apply restricted pesticides. Regression models were used to compare characteristics by race. RESULTS: Black farmers reported lower lifetime pesticide use, less use of each class of pesticides (e.g., herbicides, insecticides), less use of high exposure application methods, and fewer pesticide-related symptoms such as headaches or dizziness, skin irritation, chest discomfort and feeling nervous or depressed than did white farmers. CONCLUSIONS: Differences between black and white farmers may be explained by farm characteristics or economics. Despite lower use of pesticides, black farmers may have other work practices that affect exposure and risk. PMID- 11920963 TI - Malignant mesothelioma in Australia, 1945-2000. AB - BACKGROUND: Australia has maintained a total national malignant mesothelioma case register since 1980. There has been a marked increase in the incidence of mesothelioma in the last 20 years. Currently 450-600 cases are notified annually in a population of 20 million. While the history of the Wittenoom (Western Australia) crocidolite mine and its aftermath is well known, these cases comprise only 5% of the total. This study describes the incidence of mesothelioma in Australia from 1945 to 2000. METHODS: Using register data, time trends in mesothelioma incidence were calculated. Analyses of incidence are reported by age, sex, anatomical site, and state of notification. Associations with occupational and environmental asbestos exposure histories are described. Lung fiber content measurements were made on a subset of cases. RESULTS: Australia has had 6,329 cases of mesothelioma from 1 January 1945 to 31 December 2000. (A further 620 cases were notified in the period from 1 January 2001 to 31 October 2001.) Annual incidence rates for Australia per million population > or = 20 years (1997) were: male, 59.8; female, 10.9; total, 35.4. Incidence rates have been continually increasing and are the highest reported national rates in the world. While Western Australia has the highest rate (1997 total rate, 52.8), most cases arise from the two most populous eastern states, New South Wales and Victoria. In 88% (male 90%, female 61%) of cases, a history of asbestos exposure was obtained. Exposures occurred in a wide variety of occupational and environmental circumstances. In 80% of cases with no history of exposure, TEM lung asbestos fiber counts > 200,000 fibers > 2 microm length per gm dry lung were obtained, suggesting unrecognized exposure. CONCLUSIONS: Australia's high incidence of mesothelioma is related to high past asbestos use, of all fiber types, in a wide variety of occupational and environmental settings. The number of cases in total is expected to be about 18,000 by 2020, with about 11,000 yet to appear. PMID- 11920965 TI - Approaches to sharing occupational safety and health information on a global scale. AB - BACKGROUND: The global burden of occupational morbidity and mortality is staggering. Information sharing has been identified as a major way of reducing this burden. Past and current approaches to such sharing and application are worth examining in order to guide future efforts. METHODS: Recent literature from international agencies and others was examined to identify examples of information sharing and to determine the status of such sharing and related issues. Literature was included from the areas of surveillance, priority setting, research, dissemination, and risk management. RESULTS: Examples of global information sharing were identified and lessons were drawn from the issues attendant to them. CONCLUSIONS: Results indicate that a broad range of efforts actively promote the global distribution of occupational safety and health information. To advance global approaches to the sharing of occupational and safety and health information, it is critical to improve the opportunity and capacity to access information. Important objectives in achieving this goal are developing coherent and transparent information policies, conducting research on dissemination, adaptation, and utilization of information, and overcoming barriers to information and training. PMID- 11920967 TI - A prospective study of computer users: II. Postural risk factors for musculoskeletal symptoms and disorders. AB - BACKGROUND: Despite widespread recommendations regarding posture during computer use, associations between specific postures and musculoskeletal health are not well characterized. METHODS: Six hundred and thirty-two newly hired computer users were followed prospectively to evaluate associations between posture and neck or shoulder (N/S) and hand or arm (H/A) musculoskeletal symptoms and musculoskeletal disorders. Participants' postures were measured at entry and they reported symptoms on weekly diaries. Participants reporting symptoms were examined for specific disorders. Multivariate Cox regression models were used to estimate associations between postural variables and risk of symptoms and disorders, controlling for confounding variables. RESULTS: Keying with an inner elbow angle > 121 degrees, greater downward head tilt, and presence of armrests on the participants chair were associated with lower risk of N/S symptoms or N/S disorders. Keying with elbow height below the height of the "J" key and the presence of a telephone shoulder rest were associated with a greater risk of N/S symptoms or N/S disorders. Horizontal location of the "J" key > 12 cm from the edge of the desk was associated with a lower risk of H/A symptoms and H/A disorders. Use of a keyboard with the "J" key > 3.5 cm above the table surface, key activation force > 48 g, and radial wrist deviation of > 5 degrees while using a mouse was associated with a greater risk of H/A symptoms or H/A disorders. The number of hours keying/week was associated with H/A symptoms and disorders. CONCLUSIONS: The results suggest that the risk of musculoskeletal symptoms and musculoskeletal disorders may be reduced by encouraging specific seated postures. PMID- 11920966 TI - A prospective study of computer users: I. Study design and incidence of musculoskeletal symptoms and disorders. AB - BACKGROUND: A prospective study of computer users was performed to determine the occurrence of and evaluate risk factors for neck or shoulder (N/S) and hand or arm (H/A) musculoskeletal symptoms (MSS) and disorders (MSD). METHODS: Individuals (n = 632) newly hired into jobs requiring > or = 15 hr/week of computer use were followed for up to 3 years. At study entry, workstation dimensions and worker postures were measured and medical and psychosocial risk factors were assessed. Daily diaries were used to document work practices and incident MSS. Those reporting MSS were examined for specific MSD. Incidence rates of MSS and MSD were estimated with survival analysis. Cox regression models were used to evaluate associations between participant characteristics at entry and MSS and MSD. RESULTS: The annual incidence of N/S MSS was 58 cases/100 person years and of N/S MSD was 35 cases/100 person-years. The most common N/S MSD was somatic pain syndrome. The annual incidence of H/A MSS was 39 cases/100 person years and of H/A MSD was 21 cases/100 person-years. The most common H/A disorder was deQuervain's tendonitis. Forty-six percent of N/S and 32% of H/A MSS occurred during the first month of follow-up. Gender, age, ethnicity, and prior history of N/S pain were associated with N/S MSS and MSD. Gender, prior history of H/A pain, prior computer use, and children at home were associated with either H/A MSS or MSD. CONCLUSIONS: H/A and N/S MSS and MSD were common among computer users. More than 50% of computer users reported MSS during the first year after starting a new job. PMID- 11920968 TI - Airways inflammation after exposure in a swine confinement building during cleaning procedure. AB - BACKGROUND: Healthy volunteers exposed for 3 hr during weighing of pigs develop an airway inflammation characterized by a massive influx of neutrophilic granulocytes in the upper and lower airways and increased bronchial responsiveness to methacholine. The purpose of the present study was to investigate health effects from exposure during cleaning of the swine confinement building and to evaluate the effect of a respiratory protection device. METHODS: Sixteen subjects were exposed for 3 hr during cleaning of a swine confinement room with a high-pressure cleaner. Seven out of sixteen subjects were equipped with a mask during exposure. RESULTS: The bronchial responsiveness increased in all subjects following exposure, significantly more in the group exposed without a mask (P < 0.05). The cell concentration (mainly neutrophilic granulocytes) in nasal lavage fluid as well as the concentration of interleukin-8, increased significantly only in those subjects exposed without a respiratory protection device. In peripheral blood, an increase of neutrophilic granulocytes was observed in both groups, although it was significantly higher in the group without mask (P < 0.05). The inhalable dust level was 0.94 (0.74 - 1.55) mg/m(3) and respirable dust 0.56 (0.51-0.63) mg/m(3). CONCLUSION: Exposure to dust aerosols during the cleaning of the interior of a swine confinement building induces increased bronchial responsiveness and an acute inflammatory reaction in the upper airways. The use of a mask attenuated but did not abolish the inflammatory response. This suggests that gases and/or ultrafine particles in this environment could be important factors in the development of increased bronchial responsiveness. PMID- 11920969 TI - Interstitial pulmonary fibrosis after severe exposure to welding fumes. AB - BACKGROUND: Interstitial pulmonary fibrosis (IPF) is reported after long term, severe exposure to welding fumes in poorly ventilated workplaces. METHODS: Fifteen welders with IPF were examined--13 in our outpatient clinic--from 1990 to 1997. Occupational histories and examinations, lung function analyses, symptoms and clinical findings, histological analyses in 13 patients partly including SEM/EDX-analyses, chest X-rays, chest computed tomographies were conducted. RESULTS: Duration of work as welders was 28 years and the cumulative dose of welding fumes 221 mg/m(3) x years (median). Lung function studies found pattern of restriction or combined restriction-obstruction, lower diffusion capacity, and reduced blood oxygen tension at exercise. Histologically, patchy interstitial fibrosis was noted. Accumulations of particulate matter typically for welding fume were detected. EDX showed increase of iron load and close topographical relationship to welding fume particles embedded in areas of scattered fibrosis. CONCLUSION: While epidemiological data are limited, it is reasonable to conclude that a causal relationship exists between IPF in welders with long term exposure to high concentrations of welding fumes. PMID- 11920970 TI - Grouping participants for tailored work-site weight loss programs: a signal detection approach. AB - BACKGROUND: Although work-site obesity prevention programs have been widely promoted, they have a high rate of attrition and participants find it very difficult to maintain the decreased weight. It is necessary to develop effective work-site programs that match the type of intervention to the participants and offer the necessary support. To this end, higher-order interaction of the causal factors of obesity needs to be analyzed. METHODS: The subjects were male, white collar workers (20-64 years of age), in Osaka, Japan. Since conventional methods, such as regression analysis or analysis of variance, cannot deal with the interaction of many variables, signal detection analysis by Kraemer was used to identify the higher-order interaction of multiple predictors of obesity. RESULTS: Out of 15 independent variables, a higher-order interaction consisting of 8 significant variables was identified. Consequently, the subjects were categorized into nine subgroups. It was revealed that the obesity of two groups of workers, 40 or more years old with a high degree of obesity, had different causes: one was related to working conditions, and one was related to smoking cessation. For the other terminal groups, further factors related to obesity were revealed. CONCLUSION: Although the applicability of the findings is limited, the methodology using signal detection analysis might be applicable to other weight loss programs as a way of facilitating the matching of the type of intervention and the target group. PMID- 11920971 TI - Hospitalization experiences of U.S. servicemembers before, during, and after participation in peacekeeping operations in Bosnia-Herzegovina. AB - BACKGROUND: There are relationships among morbidity experiences before, during, and after participation in overseas military operations. METHODS: U.S. servicemembers who deployed to Bosnia-Herzegovina during a 4-year period were classified based on their last hospitalizations prior to deploying. Hospitalization rates during and following deployment were calculated in relation to the timing and causes of pre-deployment hospitalizations. RESULTS: Deployers ever hospitalized pre-deployment were 120% and 50% more likely to be hospitalized during and following deployment, respectively. For nearly every category of diagnoses, hospitalization rates during and following deployment were highest among those hospitalized for the same category, intermediate among those hospitalized for other categories, and lowest among those not hospitalized prior to deploying. Deployers hospitalized within 1 month, 2-3 months, or > 3 months of deploying were 3.8, 2.6, and 1.4-times more likely to be hospitalized during deployment. CONCLUSIONS: The nature and recency of prior hospitalizations significantly determine during and post-deployment hospitalization risks. PMID- 11920972 TI - Poisoning as a result of barium styphnate explosion. AB - BACKGROUND: Acute barium poisoning is most often the result of accidental or suicidal ingestion of the rodenticide, barium carbonate. We describe a trauma patient whose condition was complicated by severe acute barium toxicity from an explosion of the propellant, barium styphnate. In addition to critical injuries, the patient manifested classic signs of barium toxicity including repeated profound hypokalemia, cardiac arrhythmias, respiratory failure, prolonged gastrointestinal dysfunction, paralysis, myoclonus, hypertension, and profound lactic acidosis. METHODS: The patient required lidocaine for ventricular bigeminy, massive infusions of potassium, prolonged ventilatory support, and parenteral nutrition to manage the effects of barium toxicity. RESULTS: He is the first reported case to demonstrate recurrent profound hypokalemia as an effect of blood transfusions. CONCLUSIONS: Considering the paucity of information concerning management of this life-threatening problem, the pathophysiology of barium toxicity, including the transfusion related hypokalemia, and its management is reviewed. PMID- 11920973 TI - Day hospitalization programs for eating disorders: a systematic review of the literature. AB - OBJECTIVE: Partial day hospitalization programs for the treatment of eating disorders are increasingly being developed worldwide. METHOD: First, a systematic database search of day hospitalization programs for eating disorders, published in either English or German, was conducted. Programs that provided sufficient information on their program structure were summarized and compared across various dimensions. Second, the responsible program directors were contacted to provide additional information regarding outcome data, current trends, challenges, and future directions of their programs. Third, outcome data from day programs presented at international conferences were included to expand the base of the review. DISCUSSION: Although the programs from different countries and health care environments varied in terms of their purpose and operated within very different health care systems, many similarities were found to exist, including the use of a multidisciplinary staff and reliance on group treatment as the primary means of therapy. Marked differences were noted in terms of inclusion criteria and intensity of care. PMID- 11920974 TI - Does environment matter? A review of nonshared environment and eating disorders. AB - OBJECTIVE: Nonshared environmental influences are experiences that are unique to siblings reared in the same family. We review studies highlighting the importance of nonshared factors for the development of eating disorders and suggest areas for future research. METHODS: Findings from behavioral genetic studies of eating disorders as well as methodological issues are reviewed. RESULTS: Twin studies suggest that approximately 17%-46% of the variance in both anorexia nervosa (AN) and bulimia nervosa (BN) can be accounted for by nonshared environmental factors. Studies directly examining these influences are scarce, although initial data indicate that differential paternal relationships, body weight teasing, peer group experiences, and life events may account for the development of eating pathology in one sibling versus another. DISCUSSION: Additional research is needed to identify specific nonshared environmental influences on eating disorders such as differential parental and sibling treatment, disparate peer group characteristics, and differential experience of life events such as physical and sexual abuse. PMID- 11920975 TI - A meta-analytic examination of the relationship between child sexual abuse and eating disorders. AB - OBJECTIVE: This study had two goals. The first was to assess the magnitude and consistency of the relationship between child sexual abuse (CSA) and eating disorders (ED). The second was to examine methodological factors contributing to the heterogeneity of this relationship. METHOD: Meta-analysis was used to examine both questions. Fifty-three studies were included in the analysis. RESULTS: A small, significant positive relationship between CSA and ED emerged. The relationship was marked by heterogeneity. Effect sizes were largest when CSA was the grouping variable, the Eating Disorders Inventory (EDI) or the Eating Attitudes Test (EAT) was used as the measure of eating disorders, and nonclinical groups were compared with clinical samples. DISCUSSION: Models of CSA and ED need to more clearly specify what aspects of ED (e.g., body image or binge eating) are most influenced by which types of CSA. These specific relationships then need to be examined empirically. PMID- 11920976 TI - Long-term impact of treatment in women diagnosed with bulimia nervosa. AB - OBJECTIVE: Both cognitive-behavioral therapy (CBT) and antidepressant medication have demonstrated efficacy in the treatment of bulimia nervosa. However, data concerning the long-term impact of such treatments have been limited. This study sought to determine if treatment with CBT and antidepressant medication was associated with better long-term outcome among women diagnosed with bulimia nervosa. METHOD: Women (N = 101) who completed a controlled treatment study of bulimia nervosa participated in follow-up assessments approximately 10 years later. RESULTS: Women who received treatment with CBT or antidepressant medication or both reported improved social adjustment at long-term follow-up compared with women randomized to the placebo condition. DISCUSSION: Treatments with demonstrated efficacy for short-term outcome appear to improve psychosocial function at long-term follow-up among women initially diagnosed with bulimia nervosa. PMID- 11920977 TI - A preliminary controlled evaluation of an eating disturbance psychoeducational intervention for college students. AB - OBJECTIVE: Because conventional preventive interventions have had little success in reducing eating pathology, we developed and evaluated a more intensive psychoeducational intervention. METHOD: Female college students who underwent this intervention and a matched control sample of students (N = 66) completed pretest and posttest surveys. RESULTS: Intervention participants showed significant decreases in thin-ideal internalization, body dissatisfaction, dieting, eating disorder symptoms, and weight over the 4-month study period, whereas matched control participants did not show changes in these outcomes with the exception that they gained weight. DISCUSSION: These preliminary findings suggest that this intervention may prove useful in reducing eating disturbances and overweight among college students, as well as the risk factors for this serious mental and physical health problem. PMID- 11920978 TI - Exercise augments the effects of cognitive-behavioral therapy in the treatment of binge eating. AB - OBJECTIVE: To evaluate the effects of adding exercise and maintenance to cognitive-behavior therapy (CBT) for binge eating disorder (BED) in obese women. METHOD: One hundred fourteen obese female binge eaters were randomized into four groups: CBT with exercise and maintenance, CBT with exercise, CBT with maintenance, and CBT only. RESULTS AND DISCUSSION: Eighty-four women completed the 16-month study. Subjects who received CBT with exercise experienced significant reductions in binge eating frequency compared with subjects who received CBT only. The CBT with exercise and maintenance group had a 58% abstinence rate at the end of the study period and an average reduction of 2.2 body mass index (BMI) units (approximately 14 lb). BMI was significantly reduced in the subjects in both the exercise and maintenance conditions. The results suggest that adding exercise to CBT, and extending the duration of treatment, enhances outcome and contributes to reductions in binge eating and BMI. PMID- 11920979 TI - Body image treatment within an inpatient program for anorexia nervosa: the role of mirror exposure in the desensitization process. AB - OBJECTIVE: This pilot study examined the role of mirror confrontation in the desensitization process of a body image treatment within an inpatient program for anorexia nervosa. METHOD: A within-subjects design was used. It compared the impact of two modes of group body image treatment on body dissatisfaction, anxiety, and avoidance behaviors. Both treatments followed a set format of exposure exercises and homework, but the modified treatment also included mirror confrontation as an exposure exercise. RESULTS: Standard treatment did not produce any significant changes. Modified treatment produced a significant and sustained improvement in body dissatisfaction and a significant reduction in body anxiety and avoidance behaviors. DISCUSSION: Mirror confrontation is a more effective form of exposure because of the strong emotional response it elicits. Patients' pronounced emotional response to this exercise allowed easier identification of the affective and behavioral components of body dissatisfaction and more cogent links into a developmental body image timeline. PMID- 11920980 TI - Longitudinal comparison of anorexia nervosa subtypes. AB - OBJECTIVE: To compare patients with restricting anorexia nervosa (ANR) and binge/purge anorexia nervosa (ANBP) on measures of impulsivity, course, and outcome. METHODS: One hundred thirty-six treatment-seeking women with AN followed prospectively for 8-12 years were reclassified at intake as 51 ANRs and 85 ANBPs according to the DSM-IV subtyping classification. Lifetime Axis I and Axis II disorders were assessed using structured interviews; follow-up interviews were conducted at 6-12-month intervals to collect weekly data on eating disorder symptomatology. RESULTS: Women with ANR and ANBP did not differ on history of substance abuse, kleptomania, suicidality, or borderline personality diagnosis at intake, or on rates of recovery, relapse, or mortality. By 8 years of follow-up, 62% of women with ANR crossed over to ANBP prospectively and only 12% of women with AN never reported regular binge/purge behaviors. CONCLUSIONS: The findings on impulsivity, course, and outcome do not support the current subtyping system. The high crossover rate in our sample from ANR to ANBP suggests that ANR represents a phase in the course of AN rather than a distinct subtype. PMID- 11920981 TI - Eating disorder symptomatology is not associated with pregnancy and perinatal complications in a cohort of adolescents who were born preterm. AB - OBJECTIVES: We attempt to resolve the question of whether pregnancy complications and perinatal trauma, including brain insults, in premature infants increase the susceptibility to eating disorder symptomatology during the adolescent years. METHOD: This study uses a historical, prospective methodology to investigate the 84 members of a cohort of infants born prematurely (<33 weeks gestation) at Thomas Jefferson University Hospital during a 25-month period, from 1979 to 1981. We extracted the following information from their neonatal intensive care unit (NICU) records: ultrasound examination findings (graded on intraventricular hemorrhage [IVH] and periventricular leukomalacia [PVL]), records of pregnancy complications and perinatal trauma, and medical problems during the NICU stay. This method eliminated recall bias, a problem with previous studies. We followed up the members of this cohort, obtaining data on 53 (63%). We correlated the NICU data with the following outcome measures: physical measurements and psychosocial, psychological, and eating disorder symptomatology data obtained by self-report questionnaires. Pregnancy and perinatal complications were combined into one composite variable. We used the method of multiple discriminant function analysis to determine statistical significance between groups. RESULTS: There were no statistically significant differences between the low (0-2) and high (3-7) composite variable of pregnancy/perinatal complications and outcome variables. CONCLUSIONS: These results indicate that traumatic episodes early in life, including brain insults, do not appear to increase the susceptibility of developing eating disorder symptomatology, depression, deficiency of self-esteem, or distortion of body shape during late adolescence. PMID- 11920982 TI - Empirical support for a model of dieting: findings from structural equations modeling. AB - OBJECTIVE: This paper presents the results of an evaluation of a model of dieting (Huon, G.F., & Strong, K.G., International Journal of Eating Disorders, 23, 361 370, 1998). It represents the culmination of a large-scale time-extended study of dieting among adolescent girls. METHODS: Data were collected from approximately 1,000 girls. A battery of questionnaires assessed dieting status, social influence, vulnerability (conformity) disposition, protective social skills, and aspects of the familial context as core components of the model. RESULTS: When the data were subjected to analyses within structural equations modeling, all specific hypotheses within the model found strong support. Moreover, multiple indices revealed that the model had a very good fit with the data and accounted for 89% of the variance in commitment to dieting. CONCLUSION: This study provides strong support for the validity of Huon and Strong's model of dieting among Australian girls. Its generality among girls in other cultures remains to be established. PMID- 11920983 TI - Correlates of subjective and objective binge eating in binge-purge syndromes. AB - OBJECTIVE: Research has begun to challenge the idea that a large amount of food is a diagnostically relevant distinction in classifying binge eating. This study examined the relationship between both objective and subjective (i.e., feeling out of control while eating an appropriate amount of food) binge eating frequency and factor analytically derived measures of dieting and psychopathology. METHOD: Participants were 40 women who were admitted for inpatient treatment for bulimia nervosa, anorexia nervosa, binge-purge type, or eating disorder not otherwise specified with binge-purge symptoms. RESULTS: Controlling for subjective binge frequency, neither the psychopathology nor the dieting factors were related to objective binge frequency. Controlling for objective binge frequency, dieting, but not psychopathology, was related to subjective binge frequency. There was no correlation between frequency of subjective and objective binge eating. DISCUSSION: The results are discussed in light of research on restrained eating. Implications for diagnosis and the role of loss of control in defining binge behavior are also discussed. PMID- 11920984 TI - Prevention of eating disorders: challenges and opportunities. AB - OBJECTIVE AND METHODS: On April 25, 2000, the National Institute of Mental Health (NIMH) convened a Roundtable on the Prevention of Eating Disorders to review the state of prevention science in eating disorders and formulate recommendations regarding future steps to be taken in this area of research. RESULTS AND DISCUSSION: This report summarizes the roundtable discussion. The discussion focused on four major areas: the state of the art of risk factors research, translational research, prevention research in related fields, and cutting-edge efforts in eating disorder prevention. CONCLUSIONS: The report concludes with specific recommendations. PMID- 11920985 TI - Cue reactivity as a predictor of outcome with bulimia nervosa. AB - The present study sought to evaluate specific hypotheses concerning the relation between cue reactivity and outcome among women with bulimia nervosa. Participants were 135 women aged between 17 and 45 years with a current, primary diagnosis of bulimia nervosa who participated in a randomized clinical trial evaluating the additive efficacy of exposure and nonexposure-based behavior therapy, to a core of cognitive behavior therapy (CBT). Physiological, self-report, and behavioral measures of cue reactivity to individualized high-risk binge foods were obtained at pretreatment and posttreatment. Primary, secondary, and tertiary outcome measures are reported for posttreatment and six-month follow-up. Self-report measures of cue reactivity at posttreatment were significantly positively associated with symptomatology at posttreatment. Cue reactivity at posttreatment was significantly positively associated with symptomatology at 6-month follow-up. However, cue reactivity at posttreatment did not contribute to the prediction of outcome at follow-up over and above posttreatment outcome. The notion that pretreatment cue reactivity may predict which treatment modality will be most beneficial (exposure or nonexposure-based treatment), as measured by reductions in symptomatology at posttreatment could not be supported. Implications for future research are discussed. PMID- 11920986 TI - Estimating readiness for change in anorexia nervosa: comparing clients, clinicians, and research assessors. AB - OBJECTIVE: This research compared the relative ability of clients, clinicians, and research assessors in estimating readiness for change in individuals with anorexia nervosa. METHOD: Fifty-six individuals with a current or past diagnosis of anorexia nervosa made ratings of the extent to which they perceived themselves to be ready for treatment and recovery. Clinicians and research assessors made the same ratings based on their impressions following clinical and research assessments, respectively. The outcome variables included questionnaire measures of change activities, assigned behavioral tasks, and clients' decision to accept intensive treatment. RESULTS: While research assessor and client ratings predicted questionnaire recovery activities, only research assessor ratings predicted completion of behavioral tasks and clients' decision to accept intensive treatment. Clinician ratings were not related to any of the questionnaire or behavioral recovery activity measures. DISCUSSION: Conditions favoring the accurate prediction of readiness for treatment and recovery are discussed, and implications for clinical practice are addressed. PMID- 11920987 TI - Self-esteem, personality, and eating disorders: baseline assessment of a prospective population-based cohort. AB - OBJECTIVE: To study the relationship of self-esteem and personality factors with eating disorders (ED). METHODS: A region-wide representative sample of 2862 girls 12-21 years old from Navarre (Spain) participated in the baseline assessment of a prospective study. A two-stage procedure was used, consisting in a first screening phase followed by a psychiatrist interview (DSM-IV criteria). Multivariable logistic regression models were used to examine the association of self-esteem (36-item scale) and personality characteristics (Eysenck inventory) with psychiatrist-diagnosed ED while controlling for potential confounders. RESULTS: Strong associations for ED were found with low self-esteem (adjusted odds ratio [adjOR] for the lowest quartile: 7.98, 95% CI: 3.4-18.8) and high levels of neuroticism (adjOR for the highest quartile: 9.49, 95% Cl: 3.7-24.5). DISCUSSION: Our results, although based on a cross-sectional design, support the potential role of neuroticism and low self-esteem in the onset of ED. PMID- 11920988 TI - Attentional biases and frontal functioning in anorexia nervosa. AB - OBJECTIVE: This study deals with executive functions such as abstraction, use of strategy, and response flexibility (focused attention) seen in patients with anorexia nervosa. The aim was to highlight any correlation between neuropsychological functioning and the degree of pathology for food style, body image, and physical state. METHOD: A modified Stroop test, Wisconsin Card Sorting test, Body Shape Questionnaire, and Eating Disorder Inventory-2 were administered to 20 anorexia nervosa restricters (AN-R) and 20 control subjects. RESULTS: The AN-R subjects showed nonspecific attentional biases, in addition to disorders regarding abstraction and flexibility of thought, similar to those seen in patients with obsessive-compulsive disorder. The disturbance of body image showed a direct correlation to frontal alterations. DISCUSSION: This evidence suggested that in the AN-R subjects the disturbances of body image might be linked to the alterations of abstraction and critical abilities and with an obsessive frontal functioning. PMID- 11920989 TI - Comorbidity of obsessive-compulsive disorders and duration of eating disorders. AB - OBJECTIVE: This study explored the comorbidity of obsessive-compulsive disorder (OCD) and eating disorders (ED) and examined the relationship between OCD comorbidity and the duration of ED. Subjects with OCD were expected to show a longer history of ED. METHOD: ED females (N = 237; 84 with anorexia nervosa and 153 with bulimia nervosa) were assessed using the Structured Clinical Interview for DSM-IV Axis I disorders (SCID-1). Data were analyzed by logistic regression to determine whether OCD comorbidity is associated with duration of ED. RESULTS: The total sample showed a prevalence rate for OCD of 29.5%. OCD prevalence did not differ between anorexic and bulimic subjects. OCD comorbidity was significantly associated with a longer history of ED. CONCLUSION: The findings of the study support previous research indicating a high comorbidity of ED with OCD. The results also suggest that OCD may play a role in the course of ED. Prospective studies are necessary to examine this hypothesis further. PMID- 11920990 TI - Personality, perfectionism, and attitudes toward eating in parents of individuals with eating disorders. AB - OBJECTIVE: This study compares personality and eating-related traits in parents of probands with eating disorders, with age-band matched healthy normal controls. METHOD: Data were abstracted from an international genetic study of anorexia nervosa. Information was available for the Multidimensional Perfectionism Scale (MPS), the Eating Disorders Inventory (EDI), and the Temperament and Character Inventory (TCI). Comparisons were done by multivariate analysis of variance. RESULTS: Mothers of probands showed elevated levels of perfectionism on the MPS and more concerns about weight and shape on the EDI compared with controls. Mothers who had daughters with diagnoses other than the restricting subtype of anorexia nervosa showed elevated levels of perfectionism on the MPS. CONCLUSION: These data are compatible with the notion that some personality traits, such as perfectionism, and weight and shape concerns may cluster in families of probands with eating disorders. PMID- 11920991 TI - Homosexuality as a specific risk factor for eating disorders in men. AB - OBJECTIVE: The current study examined whether homosexuality is a specific risk factor for disordered eating in men. METHOD: Men (64 heterosexual and 58 homosexual) completed the Beck Depression Inventory (BDI), the Rosenberg Self Esteem Scale (RSE), the Masculinity and Femininity scales of the Bem Sex-Role Inventory (BSRI), the Bulimia Test-Revised (BULIT-R), the Eating Attitudes Test (EAT-26), and the Body Shape Questionnaire (BSQ). RESULTS: Homosexual men had more pathological scores on the BDI, RSE, BULIT-R, EAT-26, and BSQ. Additionally, homosexual men reported greater discomfort with sexual orientation. After controlling for differences in depression, self-esteem, and comfort with sexual orientation, sexual orientation continued to account for significant variance in BULIT-R, EAT-26, and BSQ scores. DISCUSSION: Future research may benefit from exploring aspects of homosexuality that may contribute specifically to risk for disordered eating in men. PMID- 11920992 TI - Exercise status and sexual orientation as moderators of body image disturbance and eating disorders in males. AB - OBJECTIVE: This study evaluated the moderating effects of sexual orientation and exercise status on measures of body image and eating disturbance in a sample of men. METHOD: One hundred and thirty-four men completed measures designed to index a range of body image facets (ideals, actual, partner's preference, overall dissatisfaction), eating disturbance (restrictive and bulimic levels), and overall self-esteem. RESULTS: There were few significant effects as a function of exercise status or sexual orientation. Bodybuilders were more satisfied on a global measure of body image and had a higher ideal and actual upper torso size rating. Gay men had a smaller actual upper torso rating than heterosexual men, but ideal size upper torso did not differ as a function of sexual orientation. DISCUSSION: Findings are discussed in terms of methodological limits of previous work and future examinations of moderating factors for body image and eating disturbance in men. PMID- 11920993 TI - Participation in aesthetic sports and girls' weight concerns at ages 5 and 7 years. AB - OBJECTIVE: The relationship between participation in aesthetic, or appearance oriented, sports and weight concerns was assessed among young girls. METHOD: Participants were part of a larger longitudinal study and included 197 and 192 girls and their mothers when girls were 5 and 7 years, respectively. At each age, girls' weight concerns and sport participation were assessed and girls were classified as participating in aesthetic sports (dance, gymnastics, cheerleading, baton twirling, swimming, aerobics, figure skating), non-aesthetic sports (volleyball, soccer, basketball, softball, hockey, tennis, martial arts, track) or no sports. RESULTS: Girls in the aesthetic sport group reported higher weight concerns than girls in the nonaesthetic and no sport groups at ages 5 and 7 years. In addition, girls participating in aesthetic sports at ages 5 and 7 years reported the highest weight concerns at age 7. DISCUSSION: Participation in aesthetic, or appearance-oriented, sports may foster a heightened focus on weight and body shape among girls. PMID- 11920994 TI - The relationship of self-esteem and body esteem in women with and without eating disorders. AB - OBJECTIVE: We investigated the predictive utility of three dimensions of body esteem in explaining self-esteem in samples of eating-disordered (ED) and nonclinical women. METHOD: Participants comprised 74 ED women and 103 female university students. All women completed questionnaires measuring body esteem and self-esteem and all provided information on height and weight. We ran hierarchical multiple regression analyses with self-esteem as the outcome variable and participant status (clinical or non), body mass index (BMI), body esteem, and interaction terms as predictors. RESULTS: The main finding was a Body Esteem x Participant Status interaction involving the attribution dimension of body esteem. Specifically, among ED women, the higher their body esteem attribution (i.e., the more they endorsed items such as "I think my appearance would help me get dates"), the higher their self-esteem; whereas this relationship was not found among the students. DISCUSSION: Our results contribute to an emerging profile of ED patients as highly socially sensitive. PMID- 11920995 TI - Development of the body checking questionnaire: a self-report measure of body checking behaviors. AB - OBJECTIVE: The purpose of this study was to develop a brief self-report inventory to assess body checking behaviors. METHOD: Using exploratory and confirmatory factor analyses, the 23-item Body Checking Questionnaire (BCQ) was developed. The BCQ measures the global construct of body checking behaviors with three correlated subfactors that assess checking related to overall appearance, checking of specific body parts, and idiosyncratic checking rituals. RESULTS: The BCQ was found to have good test-retest reliability (.94) and the subfactors had good internal consistency (.88, .92, and .83). The measure correlated highly with other measures of negative body image and eating disorders, demonstrating its concurrent validity. Additionally, the BCQ was found to differentiate normal controls and eating disorder patients, as well as nonclinical participants scoring high and low on a measure of concern with body size and dieting. DISCUSSION: Due to the potential role of ritualistic body checking in the maintenance of body dissatisfaction by directing excessive attention to body shape/weight, the BCQ may prove to be a useful clinical tool in the assessment and treatment of eating disorder patients. PMID- 11920996 TI - The media's representation of the ideal male body: a cause for muscle dysmorphia? AB - OBJECTIVE: This study sought to examine the effects of media images on men's attitudes toward their body appearance. METHOD: A group of college men viewed advertisements showing muscular men, whereas a control group viewed neutral advertisements. Immediately thereafter, participants performed a computerized test of body image perception while unaware of the hypotheses being tested in the study. RESULTS: The students exposed to the muscular images showed a significantly greater discrepancy between their own perceived muscularity and the level of muscularity that they ideally wanted to have. DISCUSSION: These findings suggest that media images, even in a brief presentation, can affect men's views of their bodies. PMID- 11920997 TI - Women's perceptions of the outcome of weight loss diets: a signal detection approach. AB - OBJECTIVE: Evolutionary psychology suggests that body shape is as important as body size and that, in women, certain body shapes are considered more attractive, specifically a waist-to-hip ratio (WHR) of about 0.70. Research has shown that the WHR does not change as a result of weight loss diets although it may be that women who diet do not appreciate this. We hypothesized that women would report diet outcomes that included shape change. This was investigated using a signal detection paradigm. METHOD: Two groups of female participants (high and low WHRs) were presented with images with high and low WHRs and were asked to choose images they could resemble through dieting. RESULTS: Both groups of women selected low WHR images as the outcome of their diets, supporting theories of evolutionary psychology. DISCUSSION: We conclude that women in the high WHR group may find adherence to diets problematic because the desired change in shape does not occur. PMID- 11920998 TI - An examination of willingness to self-disclose in women with bulimic symptoms considering the context of disclosure and negative affect levels. AB - OBJECTIVE: This study explored the relationship between bulimic symptoms and self disclosure. It examined whether women who reported greater bulimic symptomatology were generally less willing to self-disclose in intimate relationships or whether reluctance to disclose was confined to eating and weight concerns. METHOD: Women with high and low scores on the Bulimia Test Revised (BULIT-R) were compared on self-disclosure about three topics (daily activities, eating, and weight) and sexual and relationship issues to three targets (mother, friend, and partner). RESULTS: There were significant main and interaction effects for BULIT-R score, target, and topic of self-disclosure. When depression was controlled for, it partially explained the association between bulimic behavior and self-disclosure. DISCUSSION: Results supported a contextual model of self-disclosure. Compared with nonbulimic women, women with bulimic symptoms were less willing to self disclose certain topics to particular targets. Levels of depression explained low willingness to disclose on topics unrelated to eating and weight to some targets. PMID- 11920999 TI - Autism spectrum disorders and low body weight: is there really a systematic association? AB - OBJECTIVE: To examine the relationship between autism spectrum disorders and low body weight. METHOD: The effect of maladaptive social and communicative behavior as well as stereotyped features on the normative body mass index (BMI) was analyzed in 103 subjects with autism or Asperger syndrome. Statistics were controlled for medication, neurological signs, overactivity, and general intelligence. RESULTS: Twenty-eight percent of the male individuals had a BMI in the fifth percentile or below. Except for hyperactive behavior, none of the predictors showed a significant association with BMI. None of the subjects met diagnostic criteria for anorexia nervosa. DISCUSSION: Although low body weight is often present in male subjects with autism or Asperger syndrome, results indicate that this link is inconsistent and partly mediated by hyperactivity. The co occurrence of autism spectrum disorders and anorexia nervosa is probably due to chance. PMID- 11921000 TI - Imagery rescripting within cognitive behavior therapy for bulimia nervosa: an illustrative case report. AB - OBJECTIVE: This case report describes the use of imagery rescripting as an adjunct to cognitive behavior therapy (CBT) for bulimia nervosa. METHODS: The patient was a 22-year-old woman, who had a 6-year history of diagnosable eating problems, with no comorbid diagnosis. She had a history of emotional abuse by her family. The single session of imagery rescripting was aimed at effecting change in the core beliefs that were hypothesized to result from that emotional abuse, and hence in the associated bulimic psychopathology. RESULTS: While eight sessions of conventional CBT reduced symptom behaviors by 50%, one session of imagery rescripting led to an almost complete cessation of the remaining binge purge behaviors. This effect was maintained at 14 weeks post-imagery. CONCLUSIONS: Imagery rescripting appears to be a valuable adjunct to conventional CBT in working with cases where there is an early history of trauma. Implications for treatment of eating disorders using this technique are considered, and relevant research designs are discussed. PMID- 11921002 TI - Invited talk: 20 years of Burwalls. PMID- 11921003 TI - Does EBM offer the best opportunity yet for teaching medical statistics? AB - Trends in the teaching of statistics to medical undergraduates since the 1970s are traced. I conclude that the aim of enabling doctors to evaluate statistical information relevant to their practice preceded the arrival of evidence-based medicine (EBM). However this aim was not well realized before 1990. EBM has given an enormous opportunity for statisticians to teach critical appraisal and to orientate future doctors towards evidence-based practice. This advantage should outweigh any disadvantage of loss of our discipline's identity in modern curricula. PMID- 11921006 TI - Teaching hypothesis tests--time for significant change? AB - Confusion in the teaching of statistical inference dates back to the conflict of Fisher's P-values and significance tests with the Neyman-Pearson hypothesis testing approach. To avoid the well-known pitfalls arising from over-reliance on significance tests and the division of results into 'significant' or 'not significant', many medical journals now insist that presentation of statistical analyses includes confidence intervals as well as or instead of P-values. The confusion over how to report statistical analyses which is evident in the recent medical literature is matched by divergent teaching of hypothesis tests between the 16 U.K. medical schools represented at the April 2000 Burwalls meeting. Suggested guidelines for the teaching of statistical inference to medical students are presented, and possible future developments are discussed. PMID- 11921009 TI - Medical students' perspective on the teaching of medical statistics in the undergraduate medical curriculum. AB - Two undergraduate medical students at the University of Bristol commented on their experiences of learning medical statistics. In general, medical students' focus is on acquiring skills needed to practice clinical medicine, and great care must be taken to explain why disciplines such as statistics and epidemiology are relevant to this. Use of real examples and an emphasis on the need for evidence has meant that medical students are increasingly aware of the pressure on clinicians to justify their treatment decisions, and the associated need to be able to understand and critically appraise medical research. It was felt that medical statistics courses should focus on critical appraisal skills rather than on the ability to analyse data, which can be acquired by particular students when they need to do this. Medical statistics should be taught early in the curriculum, but there is a need to reinforce such skills throughout the course. Teaching and assessment methods should recognize that what is being taught is a practical skill of clinical relevance. This means that problem based small groups, data interpretation exercises and objective structured clinical examinations will be more productive than traditional teaching and examination methods. PMID- 11921011 TI - Teaching epidemiology and statistics by distance learning. PMID- 11921013 TI - Approach to Hodgkin's lymphoma in the new millennium. AB - Approximately 75% of patients with Hodgkin's lymphoma can be cured with modern chemotherapy and radiation. Most patients are treated according to clinical stage and the associated prognostic factors. For patients with limited stage Hodgkin's lymphoma, combined modality treatment has replaced subtotal nodal irradiation as the preferred treatment option. This approach eliminates laparotomy and potentially decreases the long-term toxicity secondary to extended field irradiation and splenectomy. Furthermore, recent studies suggest that it may improve disease control and possibly survival. Multiple novel regimens have been tested in the past 20 years in patients with advanced Hodgkin's lymphoma including dose-intense regimens, but current evidence suggests that ABVD remains the treatment of choice outside clinical trials. Over the past decade, the treatment-related morbidity and mortality associated with autologous stem cell transplantation have reduced significantly and stem cell transplant is becoming the treatment of choice for most patients with primary refractory or recurrent Hodgkin's lymphoma. With longer follow-up, long-term complications, in particular secondary malignancy have become the leading cause of late treatment failure for patients with Hodgkin's lymphoma. To improve the overall outcome of patients with Hodgkin's lymphoma, future studies need to focus on reducing the therapy-related toxicity for patients with good risk disease as well as improving disease control for patients with poor risk disease through a risk-adapted approach. PMID- 11921015 TI - Abnormalities in hemostasis in acute promyelocytic leukemia. AB - Our understanding of the hemostatic abnormalities in acute promyelocytic leukemia (APL) has undergone remarkable changes over the past three decades. Hemorrhagic complications lead to significant morbidity and mortality in patients with APL. Up to 30% of early deaths are due to hemorrhagic complications. Our concept of the coagulation changes in APL have evolved from a simple belief that the bleeding was due to disseminated intravascular coagulation (DIC) from underlying infection, to a knowledge that the APL cells themselves are responsible for the bleeding. We have observed that apoptotic APL cells have increased generation of thrombin and therefore contribute to the hypercoagulability and DIC. In addition, excessive fibrinolysis has been recognized to play an important role. The advent of a new therapeutic approach with all-trans-retinoic acid has improved overall survival and has greatly shortened the hemorrhagic phase of the disease, but only to produce a new complication, the retinoic acid syndrome. Its pathogenesis and management still remain to be the challenge for the present. PMID- 11921014 TI - Transcriptional and post-transcriptional mechanisms of glucocorticoid antiproliferative effects. AB - Glucocorticoids (GCs) are used as immunosuppressive and anti-inflammatory agents in treating organ transplantation rejection, autoimmune diseases, (hematological) cancers, and inflammatory disorders. GCs exert their effects through a multitude of mechanisms, the most significant of which is inhibition of cytokine production, and for some cytokines their effects on target cells. Paradoxically, GCs also upregulate the expression of (pro-inflammatory) high-affinity cytokine receptors on target cells in the face of lost ligand (cytokine) stimulation. GC inhibition of cytokine expression occurs at both transcriptional and post transcriptional levels. GCs acted transcriptionally by binding their cytosolic receptor (GR), thereby facilitating its nuclear translocation and subsequent binding to the promoter region of cytokine genes on sites compatible with GC response element (GRE) motifs, which in turn directly or indirectly regulated gene expression. In addition to direct DNA binding, GCs acted post transcriptionally by: (1) antagonism of nuclear factors required for efficient gene expression either directly or through induction of the expression of specific transcription factor antagonists, (2) altered Th lineage development by favouring the generation of (anti-inflammatory) Th2 cells and suppressing the induction or the activity of established (pro-inflammatory) Th1 cells, and (3) stimulating the expression of transforming growth factor (TGF)-beta, an immunosuppressive cytokine which inhibited cytokine production. However, these mechanisms are not mutually exclusive, since GCs may utilize more than one mechanism in exerting their anti-proliferative effect. PMID- 11921016 TI - Recent publications in hematological oncology. PMID- 11921018 TI - Expression of tissue factor and vascular endothelial growth factor is associated with angiogenesis in colorectal cancer. AB - We examined the expression of tissue factor (TF) and vascular endothelial growth factor (VEGF) and the microvessel density (MVD) in 100 patients with colorectal cancer, and we investigated the relationship of the expression of TF or VEGF with angiogenesis. TF antigen was positive in 57.0% of all specimens. Incidence of TF expression was 41.2%, 45.5%, 52.6%, 84.6%, and 81.3% in tumors from patients in clinical stages I, II, IIIA, IIIB, and IV, respectively. TF expression was correlated with the Dukes' classification (P = 0.01) and the clinical stage of colorectal cancer (P = 0.02). VEGF antigen was positive in 64.0% of all specimens. Incidence of VEGF expression was 41.2%, 57.6%, 73.7%, 84.6%, and 75.0% in tumors from patients in clinical stages I, II, IIIA, IIIB, and IV, respectively. VEGF expression was correlated with the Dukes' classification (P = 0.01) but showed a weak association with the clinical stage (P = 0.08). MVD was significantly associated with the depth of invasion (P = 0.01), lymph node metastasis (P = 0.001), and liver metastasis (P = 0.02). The mean values of MVD were 7.5 +/- 2.8, 10.1 +/- 5.7, 14.6 +/- 5.8, 13.5 +/- 3.9, and 15.9 +/- 4.2 in tumors from patients in clinical stages I, II, IIIA, IIIB, and IV, respectively. A close relationship between VEGF and MVD (P < 0.001) and a significant correlation between TF expression and MVD were observed (P = 0.02). TF-positive carcinomas presented high MVD and VEGF expression (P < 0.001) more frequently than did TF-negative tumors. These results suggest that involvement of TF in the process of metastasis and progression of colorectal cancer may depend on increased angiogenesis. PMID- 11921017 TI - Optimization of folic acid, vitamin B(12), and vitamin B(6) supplements in pediatric patients with sickle cell disease. AB - Using homocysteine as a functional marker, we determined optimal folic acid, vitamin B(12), and vitamin B(6) dosages in 21 pediatric sickle cell disease (SCD) patients (11 HbSS, 10 HbSC; 7-16 years). Daily supplements of folic acid (400, 700, or 1,000 microg), vitamin B(12) (1, 3, or 5 U.S. 1989 RDA), and vitamin B(6) (1 or 3 U.S. 1989 RDA) were gradually increased in an 82-week dose-escalation study. Blood was taken at 9 occasions for measurements of erythrocyte (RBC) and serum folate, plasma vitamin B(12), whole-blood vitamin B(6), and plasma homocysteine. Augmentation of folic acid from 700 to 1,000 microg and vitamin B(12) from 3 to 5 RDA did not further decrease homocysteine. Percentages of patients exhibiting significant individual homocysteine decreases amounted to 43% (folic acid from 0 to 400 microg, vitamins B(12) and B(6) from 0 to 1 RDA), 14% (folic acid from 400 to 700 microg), 24% (vitamin B(12) from 1 to 3 RDA), and 18% (vitamin B(6) from 1 to 3 RDA ). The lowest plasma homocysteine at 82 weeks was 5.9 +/- 2.2 micromol/L. Patients with HbSS had higher RBC folate than HbSC. The entire group exhibited an inverse relation between RBC folate and hemoglobin. We conclude that RBC folate is less valuable for folate status assessment in SCD patients. Optimal dosages are as follows: 700 microg folic acid (3.5-7 U.S. 1989 RDA), 3 U.S. 1989 RDA vitamin B(12) (4.2-6.0 microg), and 3 U.S. 1989 RDA vitamin B(6) (4.2-6.0 mg). A practical daily combination is 1 mg folic acid (4.3-8.5 U.S. 1998 RDA when taken with meals), 6 microg vitamin B(12) (2.5-5 U.S. 1998 RDA), and 6 mg vitamin B(6) (4.6-10 U.S. 1998 RDA). This combination may by simple and relatively inexpensive means reduce these patients' inherently high risk of endothelial damage. PMID- 11921019 TI - Global fibrinolytic capacity in early respiratory distress syndrome: a pilot study. AB - Fibrin deposits found in the pulmonary microcirculation and in the small airways of preterm infants with severe respiratory distress syndrome (RDS) has been explained by the activation of the coagulation system and/or insufficient fibrinolysis. Previous studies suggest that disseminated intravascular coagulation is not prominent in the early stages of RDS, and there is a reduced fibrinolytic activity in these patients. The in vitro evaluation of body's fibrinolytic potential is difficult in contrast to blood clotting because fibrinolysis is only activated after coagulation. To solve this problem a standardized assay was developed to measure the global fibrinolytic capacity (GFC) in plasma. This assay allows us a sensitive and reliable parameter to evaluate the fibrinolytic potency of plasma in vitro. We therefore studied GFC in the first 6 hr after birth in preterm infants who later developed RDS. Global fibrinolytic capacity which is expressed as generated D-dimer concentrations was significantly lower in preterm infants who later developed RDS compared to the control group in this study. These findings support our previous hypothesis of "reduced fibrinolytic state in early RDS". PMID- 11921020 TI - Autoimmune hemolytic anemia. AB - Red blood cell (RBC) autoantibodies are a relatively uncommon cause of anemia. However, autoimmune hemolytic anemia (AIHA) must be considered in the differential diagnosis of hemolytic anemias, especially if the patient has a concomitant lymphoproliferative disorder, autoimmune disease, or viral or mycoplasmal infection. Classifications of AIHA include warm AIHA, cold agglutinin syndrome, paroxysmal cold hemoglobinuria, mixed-type AIHA, and drug-induced AIHA. Characteristics of the autoantibodies are responsible for the various clinical entities. As a result, diagnosis is based on the clinical presentation and a serologic work-up. For each classification of AIHA, this review discusses the demographics, etiology, clinical presentation, laboratory evaluation, and treatment options. PMID- 11921022 TI - Whole-pulmonary low-dose radiation therapy in agnogenic myeloid metaplasia with diffuse lung involvement. AB - Agnogenic myeloid metaplasia is a hematologic disorder accompanied by extramedullary hematopoiesis (EMH) affecting various organs. Lung involvement however is rare. We present the case of a 76-year-old woman with myelofibrosis, recurrent pleural effusions, pulmonary hypertension, and serious right cardiac failure. An open lung biopsy confirmed pulmonary EMH. She underwent low-dose (200 cGy) whole-lung radiotherapy in 4 fractions of 50 cGy each. Her clinical and hemodynamic parameters improved. We conclude that low-dose whole-lung radiation may be efficacious for the palliative treatment of pulmonary EMH. PMID- 11921021 TI - Hepatosplenic gammadelta T-cell lymphoma presenting with immune-mediated thrombocytopenia and hemolytic anemia (Evans' syndrome). AB - We describe an unusual case of hepatosplenic T-cell lymphoma in a 61-year-old man who presented with fever, hepatosplenomegaly, anemia, and thrombocytopenia. A spleen biopsy was consistent with T-cell lymphoma. Cytogenetic studies did not reveal chromosome abnormalities. Using the polymerase chain reaction approach, clonality of the T-cell receptor gamma-chain gene rearrangement could be demonstrated, while Southern blot analysis disclosed only a germline configuration of the T-cell receptor beta chain genes. Of interest, an immune mediated mechanism was demonstrated and was most likely responsible for erythrocyte and platelet destruction; this is, therefore, the first report of gamma T-cell lymphoma in association with Evans' syndrome. Initial steroid treatment was efficacious in limiting autoimmunity but constitutional symptoms did not subside. Chemotherapy (MACOP-B) was successful in obtaining complete clinical remission. Finally, thrombocytopenia in gammadelta T-cell lymphoma patients should be routinely evaluated for platelet autoantibodies. PMID- 11921023 TI - Clinical severe hepatic venoocclusive disease during induction treatment of acute monoblastic leukemia managed with defibrotide. AB - Hepatic venoocclusive disease (VOD) is the most common complication of cytoreductive therapy used for stem cell transplantation, but it is rarely encountered during induction treatment of acute leukemia. We describe a patient in whom severe clinical VOD developed shortly after induction treatment for acute monoblastic leukemia. Administration of intravenous defibrotide for 19 days induced complete resolution of the VOD. Further consolidation treatment was resumed including high-dose cytosine arabinoside without further complications. PMID- 11921024 TI - Multiple herpetic whitlow lesions in a patient with chronic lymphocytic leukemia. AB - Herpetic whitlow, a herpes simplex virus infection involving the digits, most commonly presents as a vesicular eruption involving a single digit. Diagnosis of herpetic whitlow can usually be made with the history of exposure, the characteristic vesicular eruption, and a positive Tzank smear and/or viral culture. We describe a case of herpetic whitlow in a patient finishing 6 cycles of chemotherapy for refractory chronic lymphocytic leukemia that presented with a bilateral, multi-digit, crusted eruption of the hands. This is an illustrative case of an immunocompromised host status altering appearance and course of cutaneous disease such that the history and physical exam alone may not help in diagnosing atypical presentations of herpetic infections. This case underscores the necessity for clinico-histopathologic correlation. PMID- 11921025 TI - EBV-associated lymphoma and chronic inflammatory demyelinating polyneuropathy in an adult without overt immunodeficiency. AB - Epstein-Barr virus (EBV)-associated lymphoproliferative disease occurs almost exclusively in immunocompromised states. This type of malignancy nevertheless developed in a woman without overt immunocompromise. She presented with rapidly progressive lower extremity weakness that was consistent with a chronic inflammatory demyelinating polyneuropathy (CIDP). CIDP has been documented in Hodgkin's disease but rarely in non-Hodgkin's disease. Diagnosis was confirmed by lymph node and sural nerve biopsies and by nerve conduction studies. Prednisone, rituximab, cyclophosphamide, doxorubicin, and etoposide were administered with regression of lymphadenopathy and improvement in neurologic symptoms. EBV associated lymphoproliferative disease can thus develop in the absence of overt immunodeficiency and may trigger a demyelinating polyneuropathy. PMID- 11921026 TI - Acquired factor VIII inhibitor treated with cyclophosphamide, vincristine, and prednisone. AB - Life-threatening bleeding can occur in non-hemophiliacs with factor VIII inhibitor and is difficult to control. Various methods have been used to suppress the inhibitor. Since 1993 we treated 6 non-hemophilic patients with factor VIII inhibitor with cyclophosphamide, vincristine, and prednisone (CVP) every 3-4 weeks. Five had complete response with disappearance of factor VIII inhibitor and normalization of factor VIII after 1 to 7 cycles of CVP. One patient did not respond after 4 cycles of CVP; mitoxantrone was added to additional two cycles of CVP, and the inhibitor disappeared. PMID- 11921027 TI - Interpretation of serum ferritin in anemia of chronic disease. PMID- 11921028 TI - Anemia following human parvovirus B19 infection in a patient with polycythemia vera. PMID- 11921029 TI - Short period of administration of diethylstilbestrol in stuttering priapism in sickle cell anemia. PMID- 11921030 TI - Successful cyclosporin A therapy for acquired amegakaryocytic thrombocytopenic purpura. PMID- 11921033 TI - Origin of evolutionary novelty: examples from limbs. AB - Classic hypotheses of vertebrate morphology are being informed by new data and new methods. Long nascent issues, such as the origin of tetrapod limbs, are being explored by paleontologists, molecular biologists, and functional anatomists. Progress in this arena will ultimately come down to knowing how macroevolutionary differences between taxa emerge from the genetic and phenotypic variation that arises within populations. The assembly of limbs over developmental and evolutionary time offers examples of the major processes at work in the origin of novelties. Recent comparative developmental analyses demonstrate that many of the mechanisms used to pattern limbs are ancient. One of the major consequences of this phenomenon is parallelism in the evolution of anatomical structures. Studies of both the fossil record and intrapopulational variation of extant populations reveal regularities in the origin of variation. These examples reveal processes acting at the level of populations that directly affect the patterns of diversity observed at higher taxonomic levels. PMID- 11921032 TI - Learning from history: morphology's challenges in Germany ca. 1900. AB - A century ago, Carl Gegenbaur's program of vertebrate evolutionary morphology faced its greatest challenges. The controversy over the evolutionary origin of the vertebrate paired limbs between 1875 and 1906 illustrates the failure of the traditional methods of comparative anatomy and embryology (supported by Haeckel's biogenetic law) to choose between different phylogenetic hypotheses. The controversy over morphology's status as science intensified at the turn of the twentieth century, when the legitimacy of historical explanation itself as a mode of scientific understanding came under fire. Gegenbaur's intellectual grandson, Hermann Braus, sought to defend the legitimacy of phylogenetic reconstruction while updating it to include experimental and causal-analytical approaches, but was unable to sustain a viable synthetic research program. The article concludes with reflections on approaches to the past used by historians and evolutionary morphologists. PMID- 11921034 TI - Vertebrate limb development and possible clues to diversity in limb form. AB - Chick embryos are good models for vertebrate development. The principles that underlie chick wing development have been discovered and there is increasing knowledge about the molecules involved. The importance of identifying molecules is that this provides a direct link to understanding the genetic basis of diversity in form. Chick wing development will be compared with limb development in other vertebrates. Possible mechanisms that could lead to variations in form, including limb reductions and limblessness, differences between fore- and hindlimbs, limb proportions, and interdigital webbing can be suggested. PMID- 11921035 TI - Future directions for the analysis of musculoskeletal design and locomotor performance. AB - New techniques and conceptual frameworks offer new challenges and exciting opportunities for research on the biomechanics and physiology of vertebrate musculoskeletal design and locomotor performance. Past research based on electromyography and two-dimensional kinematics has greatly advanced the field of vertebrate functional morphology. Studies using these approaches have revealed much about vertebrate structure and function and have emphasized the importance of incorporating historical and developmental constraint and ecological context. Continued use of these experimental tools, but with greater emphasis on three dimensional analysis of body movement, in combination with 3D kinetics and flow visualization of fluid movement past moving organisms, can now take advantage of the considerable advances in computing power and digital video technology. Indeed, surprisingly few detailed 3D analyses of movement for many locomotor modes and differing organisms are presently available. A challenge of 3D analyses will be to reduce the complexity of the data obtained in order to identify general principles of movement and biomechanics. New techniques and approaches for measuring muscle forces and length changes, together with activation patterns and movement, under dynamic conditions of more varied motor behavior are now also available. These provide the opportunity to study the mechanics and physiology of muscle function at greater depth and under more realistic conditions than has been previously possible. The importance of studying intact, behaving organisms under a broader range of locomotor conditions (other than steady state) and in the context of their natural environment remains a critical need for vertebrate biologists. This provides the much-needed opportunity for placing advances at more cellular and molecular levels into the context of whole organism function. Hence, studies at the organismal level remain paramount. PMID- 11921036 TI - Structure, form, and function of flight in engineering and the living world. AB - By combining appearance and behavior in animals with physical laws, we can get an understanding of the adaptation and evolution of various structures and forms. Comparisons can be made between animal bodies and various technical constructions. Technical science and theory during the latest decades have resulted in considerable insight into biological adaptations, but studies on structures, forms, organs, systems, and processes in the living world, used in the right way, have also aided the engineer in finding wider and better solutions to various problems, among them in the design of micro-air vehicles (MAVs). In this review, I discuss the basis for flight and give some examples of where flight engineering and nature have evolved similar solutions. In most cases technology has produced more advanced structures, but sometimes animals are superior. I include how different animals have solved the problem of producing lift, how animal wings meet the requirements of strength and rigidity, how wing forms are adapted to various flight modes, and how flight kinematics are related to flight behavior and speed. The dynamics of vorticity is summarized. There are a variety of methods for the determination of flight power; it has been estimated adequately by lifting-line theory, by physiological measurements, and from mass loss and food intake. In recent years alternative methods have been used, in which the mechanical power for flight is estimated from flight muscle force used during the downstroke. Refinements of these methods may create new ways of estimating flight power more accurately. MAVs operate at the same Reynolds numbers as large insects and small birds and bats. Therefore, studies on animal flight are valuable for MAV design, which is discussed here. PMID- 11921037 TI - Sequence heterochrony and the evolution of development. AB - One of the most persistent questions in comparative developmental biology concerns whether there are general rules by which ontogeny and phylogeny are related. Answering this question requires conceptual and analytic approaches that allow biologists to examine a wide range of developmental events in well structured phylogenetic contexts. For evolutionary biologists, one of the most dominant approaches to comparative developmental biology has centered around the concept of heterochrony. However, in recent years the focus of studies of heterochrony largely has been limited to one aspect, changes in size and shape. I argue that this focus has restricted the kinds of questions that have been asked about the patterns of developmental change in phylogeny, which has narrowed our ability to address some of the most fundamental questions about development and evolution. Here I contrast the approaches of growth heterochrony with a broader view of heterochrony that concentrates on changes in developmental sequence. I discuss a general approach to sequence heterochrony and summarize newly emerging methods to analyze a variety of kinds of developmental change in explicit phylogenetic contexts. Finally, I summarize a series of studies on the evolution of development in mammals that use these new approaches. PMID- 11921039 TI - Are Echiura derived from a segmented ancestor? Immunohistochemical analysis of the nervous system in developmental stages of Bonellia viridis. AB - Despite several unquestionably homologous characters with Annelida, the Echiura have generally been considered to form a distinct taxon due to apparent lack of segmentation: neither in the body cavity nor in any other structures of the adult animals have clear signs of a metameric organization been observed. However, it must be considered that this lack of segmentation could represent a secondary condition and that Echiura are derived from segmented ancestors. An immunohistochemical analysis visualized with confocal laser-scanning microscopy (cLSM) shows the development of serially repeated units in the nervous system of Bonellia viridis. This organization corresponds to the metameric ganglia typical of Annelida. Antibodies against the neurotransmitters serotonin (5 hydroxytryptamine) and FMRFamide label distinct subsets of neurons. Their perikarya are arranged in discrete repetitive units in the ventral nerve cord of different larval stages. Labeling of neurotubuli using antibodies against different tubulin isoforms provides additional support for the metameric character of the nervous system. Contrary to previous descriptions, the peripheral nerves extending from the ventral nerve cord into the body wall musculature are paired and are evenly distributed; their arrangement corresponds to that of the serotoninergic perikarya. Morphological and neurochemical differentiation of the nervous system clearly proceeds from anterior to posterior, indicating the occurrence of a posterior growth zone. The serial ganglionic organization of the nervous system of Articulata is generally regarded as having evolved in conjunction with repetitive muscular units, in particular with the formation of typical annelid segments. Therefore, these results are interpreted as an indication that Echiura are derived from segmented ancestors and thus support the systematic inclusion of the Echiura within the Annelida. PMID- 11921040 TI - Morphology of the prometamorphic larva of the spadefoot toad, Scaphiopus intermontanus (Anura: Pelobatidae), with an emphasis on the lateral line system and mouthparts. AB - We provide a detailed description of the larval morphology of the Great Basin spadefoot toad (Scaphiopus intermontanus), a species with documented morphological variability in larval structures associated with feeding. We based our findings on laboratory-raised individuals fed a herbivorous diet. We characterized the morphology of the prometamorphic larva (limited to developmental stages 37 and 38) and then related our findings to the larval ecology of the species. Based on its morphology, such as slightly depressed body, dorsally positioned eyes, anteroventrally oriented oral disc, intermediate tail fin height and slightly attenuated tail tip, relative lack of ventral neuromasts (compared to Xenopus laevis), and pigmentation banding patterns, and habits, such as selection of breeding sites by adults and larval foraging behavior, S. intermontanus can be characterized best as belonging to a (lentic-) benthic guild of anuran larvae. Nevertheless, the larvae are capable of occupying a broader array of ecological niches. Because we characterized individuals raised on a herbivorous diet, our morphological descriptions apply only to the herbivorous S. intermontanus larva (and perhaps to those larvae that are dietary generalists and may feed carnivorously only infrequently). Our findings can serve as a baseline for future morphological and developmental comparisons with the carnivorous morphological variant of this species. PMID- 11921041 TI - Chondrocranial development in larval Rana sylvatica (Anura: Ranidae): morphometric analysis of cranial allometry and ontogenetic shape change. AB - This study provides baseline quantitative data on the morphological development of the chondrocranium in a larval anuran. Both linear and geometric morphometric methods are used to quantitatively analyze size-related shape change in a complete developmental series of larvae of the wood frog, Rana sylvatica. The null hypothesis of isometry was rejected in all geometric morphometric and most linear morphometric analyses. Reduced major axis regressions of 11 linear chondrocranial measurements on size indicate a mixture of allometric and isometric scaling. Measurements in the otic and oral regions tend to scale with negative allometry and those associated with the palatoquadrate and muscular process scale with isometry or positive allometry. Geometric morphometric analyses, based on a set of 11 chondrocranial landmarks, include linear regression of relative warp scores and multivariate regression of partial warp scores and uniform components on log centroid size. Body size explains about one quarter to one-third of the total shape variation found in the sample. Areas of regional shape transformation (e.g., palatoquadrate, otic region, trabecular horns) are identified by thin-plate spline deformation grids and are concordant with linear morphometric results. Thus, the anuran chondrocranium is not a static structure during premetamorphic stages and allometric patterns generally follow scaling predictions for tetrapod cranial development. Potential implications regarding larval functional morphology, cranial development, and chondrocranial evolution in anurans are discussed. PMID- 11921042 TI - History and function of scale microornamentation in lacertid lizards. AB - Differences in surface structure (ober-hautchen) of body scales of lacertid lizards involve cell size, shape and surface profile, presence or absence of fine pitting, form of cell margins, and the occurrence of longitudinal ridges and pustular projections. Phylogenetic information indicates that the primitive pattern involved narrow strap-shaped cells, with low posteriorly overlapping edges and relatively smooth surfaces. Deviations from this condition produce a more sculptured surface and have developed many times, although subsequent overt reversals are uncommon. Like variations in scale shape, different patterns of dorsal body microornamentation appear to confer different and conflicting performance advantages. The primitive pattern may reduce friction during locomotion and also enhances dirt shedding, especially in ground-dwelling forms from moist habitats. However, this smooth microornamentation generates shine that may compromise cryptic coloration, especially when scales are large. Many derived features show correlation with such large scales and appear to suppress shine. They occur most frequently in forms from dry habitats or forms that climb in vegetation away from the ground, situations where dirt adhesion is less of a problem. Microornamentation differences involving other parts of the body and other squamate groups tend to corroborate this functional interpretation. Microornamentation features can develop on lineages in different orders and appear to act additively in reducing shine. In some cases different combinations may be optimal solutions in particular environments, but lineage effects, such as limited reversibility and different developmental proclivities, may also be important in their genesis. The fine pits often found on cell surfaces are unconnected with shine reduction, as they are smaller than the wavelengths of most visible light. PMID- 11921043 TI - Fine structural and cytochemical observations of dental epithelial cells during the enameloid formation stages in red stingrays Dasyatis akajei. AB - The fine structure and the localization of nonspecific acid phosphatase (ACPase), nonspecific alkaline phosphatase (ALPase), and calcium-dependent adenosine triphosphatase (Ca-ATPase) activities in the dental epithelial cells in tooth germs of Dasyatis akajei in the later stages of enameloid formation were investigated. Numerous invaginations of the distal cell membrane of the inner dental epithelial (IDE) cells were observed at the early stage of enameloid maturation. The invaginations contain many fine granular and filamentous substances; the lamina densa, which was thicker during the former stages, is obscure. Granules exhibiting defined ACPase activity were usually found in the IDE cells during the stages of enameloid mineralization and maturation. IDE cells are putatively involved in the removal of degenerated enameloid matrix during these stages. Marked ALPase activity was detected at the proximal and the lateral cell membranes of the IDE cells from the late stage of enameloid matrix formation to the early stage of enameloid maturation. Strong activity of Ca-ATPase was localized at the proximal and the lateral cell membranes of the IDE cells during the stages of enameloid mineralization and maturation. ALPase and Ca-ATPase activity is probably related to crystal formation in the enameloid and the removal of degenerated enameloid matrix from the enameloid. PMID- 11921044 TI - Lipotyphla limb myology comparison. AB - Fore- and hindlimb muscles were dissected in four species of Lipotyphla: the western European hedgehog Erinaceus europaeus (Erinaceidae, Erinaceinae); the moonrat Echinosorex gymnura (Erinaceidae, Hylomyinae or Galericinae); the tailless tenrec Tenrec ecaudatus (Tenrecidae, Tenrecinae); and the common European white-toothed shrew Crocidura russula (Soricidae, Soricinae). This work completely reviews the limb musculature of these walking mammals. Twelve myological characters were evaluated in order to disclose phylogenetic relationships. The cladogram obtained supported previous ones based on cranial and dental characters. This study shows that myological characters are valuable in phylogenetic analyses. PMID- 11921045 TI - Larval and metamorphic development of the foregut and proboscis in the caenogastropod Marsenina (Lamellaria) stearnsii. AB - The specialized, postmetamorphic feeding structures of predatory caenogastropods evolved by changes to an ancestral caenogastropod developmental program that generated a planktotrophic larval stage followed by a herbivorous postmetamorphic stage. As part of a program of comparative studies aimed at reconstructing these developmental changes, I studied the development of the postmetamorphic feeding system of Marsenina stearnsii using histological sections for light microscopy and scanning and transmission electron microscopy. The feeding system of this species has two very different designs during ontogeny. The larval system uses ciliary effectors to capture and ingest microalgae, whereas the juvenile/adult system includes a proboscis, jaws, and radular apparatus for predation on ascidian zooids. The postmetamorphic foregut begins to develop during the early larval phase, but the anlagen does not interfere with larval feeding because it develops as an increasingly elaborate outpocketing from the ventral wall of the larval esophagus. At metamorphosis, an opening is created in the anterior tip of the prospective, postmetamorphic buccal cavity and the margins of this opening anneal with the metamorphically remodeled lips of the larval mouth. This process exposes the jaws, which differentiate within the buccal cavity prior to metamorphosis. As a working hypothesis, I suggest that rupture of the buccal cavity to the outside at metamorphosis was selected as a mechanism to allow precocious development of jaws in species where jaws enhanced feeding performance by young juveniles. The larval esophagus of M. stearnsii appears to be completely destroyed at metamorphosis. Larval esophageal cells have distinctive apical characteristics (cilia, blebbed microvilli, stacks of lamellae within the glycocalyx) and no cells having this signature persist through metamorphosis. Development of the proboscis and proboscis sac, which begins prior to metamorphosis, conforms to previous descriptions of pleurembolic proboscis development, although an acrembolic proboscis has been ascribed to members of the Lamellaroidea. PMID- 11921046 TI - Relation between the outer cover of the egg case of Argiope aurantia (Araneae: Araneidae) and the emergence of its spiderlings. AB - To emerge from the egg case, Argiope aurantia spiderlings must penetrate a tightly woven outer cover composed primarily of large-diameter cylindrical gland fibers and small-diameter fibers, likely of aciniform gland origin. They accomplish this using enzymatic digestion and mastication to form a communal hole in the outer cover. The involvement of proteolytic enzymes in this process was demonstrated by zymography of spiderling homogenates and washes made from the edges of holes. The specific source(s) of the proteases is unknown, but histological examination of spiderling sections indicates that the digestive tract, venom glands, and gnathocoxal glands are all functioning at the time of emergence from the egg case. Observations on edges of holes indicate that spiderlings are able to solubilize the small-diameter fibers completely, but cylindrical gland fibers only partially. In the outer cover, cylindrical fibers are composed of numerous fibrils embedded within a matrix. Spiderlings appear to be unable to solubilize the fibrils, but digestion of the matrix allows the spiderlings to push the fibrils aside to create the opening. PMID- 11921047 TI - Contrast computed tomography scan in acute stroke: "You can't always get what you want but...you get what you need". PMID- 11921048 TI - Prognostic accuracy of cerebral blood flow measurement by perfusion computed tomography, at the time of emergency room admission, in acute stroke patients. AB - The purpose of this study was to determine the prognostic accuracy of perfusion computed tomography (CT), performed at the time of emergency room admission, in acute stroke patients. Accuracy was determined by comparison of perfusion CT with delayed magnetic resonance (MR) and by monitoring the evolution of each patient's clinical condition. Twenty-two acute stroke patients underwent perfusion CT covering four contiguous 10mm slices on admission, as well as delayed MR, performed after a median interval of 3 days after emergency room admission. Eight were treated with thrombolytic agents. Infarct size on the admission perfusion CT was compared with that on the delayed diffusion-weighted (DWI)-MR, chosen as the gold standard. Delayed magnetic resonance angiography and perfusion-weighted MR were used to detect recanalization. A potential recuperation ratio, defined as PRR = penumbra size/(penumbra size + infarct size) on the admission perfusion CT, was compared with the evolution in each patient's clinical condition, defined by the National Institutes of Health Stroke Scale (NIHSS). In the 8 cases with arterial recanalization, the size of the cerebral infarct on the delayed DWI-MR was larger than or equal to that of the infarct on the admission perfusion CT, but smaller than or equal to that of the ischemic lesion on the admission perfusion CT; and the observed improvement in the NIHSS correlated with the PRR (correlation coefficient = 0.833). In the 14 cases with persistent arterial occlusion, infarct size on the delayed DWI-MR correlated with ischemic lesion size on the admission perfusion CT (r = 0.958). In all 22 patients, the admission NIHSS correlated with the size of the ischemic area on the admission perfusion CT (r = 0.627). Based on these findings, we conclude that perfusion CT allows the accurate prediction of the final infarct size and the evaluation of clinical prognosis for acute stroke patients at the time of emergency evaluation. It may also provide information about the extent of the penumbra. Perfusion CT could therefore be a valuable tool in the early management of acute stroke patients. PMID- 11921049 TI - Functional recovery and neuroanatomical plasticity following middle cerebral artery occlusion and IN-1 antibody treatment in the adult rat. AB - Stroke is a prevalent and devastating disorder, and no treatment is currently available to restore lost neuronal function after stroke occurs. One unique therapy that may improve functional recovery after stroke is blockade of the neurite inhibitory protein Nogo-A with the monoclonal antibody IN-1, through enhancement of neuroanatomical plasticity from uninjured areas of the central nervous system. In the present study, we combined IN-1 treatment with an ischemic lesion (permanent middle cerebral artery occlusion) to determine the effect of Nogo-A neutralization on cortical plasticity and functional recovery. We report here that, following ischemic stroke and treatment with IN-1, adult rats demonstrated functional recovery on a forelimb-reaching task and new cortico efferent projections from the opposite, unlesioned hemisphere. These results support the efficacy of Nogo-A blockade as a treatment for ischemic stroke and implicate plasticity from the unlesioned hemisphere as a mechanism for recovery. PMID- 11921050 TI - Verbal memory in left temporal lobe epilepsy: evidence for task-related localization. AB - We explored the hypothesis that components of verbal memory are subserved by separate temporal lobe structures in patients with temporal lobe structures in patients with temporal lobe epilepsy [correction]. Uptake of 18F fluorodeoxyglucose (FDG) measured by positron emission tomography, hippocampal volume, and memory for arbitrarily and semantically related verbal paired associates were examined in 27 patients with left temporary lobe epilepsy. Scores from memory tests performed outside the scanner were regressed against normalized resting FDG uptake at each voxel. Significant regression was seen in the left perirhinal cortex (Talaraich coordinates x, y, z: -29, 10, -34; p < 0.05) for arbitrarily related word pairs. For semantically related paired associates, significant regression was present in the left inferior temporal gyrus (x, y, z: 48, -18, -24; p < 0.05). In subsequent analyses, mean FDG uptake within a spherical region of interest centered on the perirhinal peak predicted performance on both tasks. Mean FDG uptake in a region of interest centered on the inferior temporal peak made an additional, independent contribution to memory for semantically related pairs. Hippocampal volumes did not explain any additional variance in memory scores. Our findings indicate that heterogeneity in the left temporal lobe structures mediating verbal memory function, and support the view that the perirhinal cortex is an important mnemonic substrate. PMID- 11921051 TI - Flupirtine blocks apoptosis in batten patient lymphoblasts and in human postmitotic CLN3- and CLN2-deficient neurons. AB - Multiple gene defects cause Batten disease. Accelerated apoptosis accounts for neurodegeneration in the late infantile and juvenile forms that are due to defects in the CLN3 and CLN2 genes. Extensive neuronal death is seen in CLN2- and CLN3-deficient human brain as well as in CLN6-deficient sheep brain and retina. When neurons in late infantile and juvenile brain survive, they manage to do so by upregulating the neuroprotective molecule Bcl-2. The CLN3 gene has antiapoptotic properties at the molecular level. We show that the CLN2 gene is neuroprotective: it enhances growth of NT2 cells and maintains survival of human postmitotic hNT neurons. Conversely, blocking CLN3 or CLN2 expression in hNT neurons with adenoviral antisense-CLN3 or antisense-CLN2-AAV2 constructs causes apoptosis. The drug flupirtine is a triaminopyridine derivative that acts as a nonopioid analgesic. Flupirtine upregulates Bcl-2, increases glutathione levels, activates an inwardly rectifying potassium channel, and delays loss of intermitochondrial membrane calcium retention capacity. We show that flupirtine aborts etoposide-induced apoptosis in CLN1-, CLN2-, CLN3-, and CLN6-deficient as well as normal lymphoblasts. Flupirtine also prevents the death of CLN3- and CLN2 deficient postmitotic hNT neurons at the mitochondrial level. We show that a mechanism of neuroprotection exerted by flupirtine involves complete functional antagonism of N-methyl-D-aspartate or N-methyl-D-aspartate-induced neuronal apoptosis. Flupirtine may be useful as a drug capable of halting the progression of neurodegenerative diseases caused by dysregulated apoptosis. PMID- 11921052 TI - Antibody-mediated suppression of Vbeta5.2/5.3(+) T cells in multiple sclerosis: results from an MRI-monitored phase II clinical trial. AB - The objective of this study was to evaluate the safety and efficacy of the humanized antibody ATM-027 in a baseline versus treatment magnetic resonance imaging-monitored study. Expansion of Vbeta5.2/5.3(+) T cells has been demonstrated in the peripheral blood, cerebrospinal fluid, and brain lesions of MS patients. In a phase I study, ATM-027 depleted these cells in peripheral blood and, in parallel, T-cell MBP reactivity and IFN-gamma expression were reduced. We studied 59 patients with relapsing-remitting MS (47 on ATM-027 and 12 on placebo) stratified for HLA-DR2 status. Monthly intravenous injections were given for 6 months. Individual dose titration was employed to obtain depletion of the target T-cell level and downregulation of antigen receptor density as monitored by flow cytometry. Five monthly magnetic resonance imaging scans were performed before treatment to establish baseline activity, six during treatment, and three during follow-up. Additional immunological assessments were performed to elucidate the mechanism of action of ATM-027. The treatment was safe and well tolerated, inducing consistent suppression of the target cell population. During run-in, active lesions were found in 78.7% (37/47) of patients treated with ATM-027. During treatment, the median number of lesions was reduced by 33% (p = 0.13) independent of DR2 status. The corresponding volume of enhancement was 221 mm(3) at baseline, with a reduction of 10% during treatment. Decreased numbers of cells expressing interferon-gamma messenger RNA, and decreased T-cell reactivity to several myelin antigens were found in ATM-027 treated patients. In conclusion, consistent suppression of Vbeta 5.2/5.3(+) T cells was achieved. However, the effect size on magnetic resonance imaging was considerably less than the targeted 60%. PMID- 11921053 TI - Age-related disability in multiple sclerosis. AB - There is evidence that the clinical course of multiple sclerosis is age related. The present study evaluated the relationship between age and rate of disability progression in a large hospital-based cohort of definite cases of multiple sclerosis (n= 1,463). Patients were followed every 6 to 12 months for a total period of observation of 11,387.8 person-years. Expanded Disability Status Scale scores increased significantly with increasing current age and longer duration of disease (p=0.007). Median times to reach Expanded Disability Status Scale scores of 4.0 and 6.0, assessed using an extended Kaplan-Meier method with age as a categorical time-varying covariate, were significantly longer among patients aged 20 to 35 years compared with patients aged 36 to 50 and 51 to 65 years (p < 0.0001). Significant associations were observed between mean Expanded Disability Status Scale scores and age at disease onset, current age, and the interaction of age at disease onset and current age (p < 0.001). Current age had a greater effect (59% of variability in the model) on disease severity than did age at disease onset. Furthermore, a multiplicative effect on Expanded Disability Status Scale score was observed for age at disease onset and current age combined, indicating a faster rate of disease progression in older patients. In conclusion, the results of the current study demonstrate the impact of age on rate of disability progression in multiple sclerosis and suggest that an age-adjusted progression index may be a more relevant criterion for defining differences between multiple sclerosis groups. PMID- 11921054 TI - Interferon beta-1a for early multiple sclerosis: CHAMPS trial subgroup analyses. AB - The objective of this work was to assess the effect of interferon beta-1a (Avonex) on the rate of development of clinically definite multiple sclerosis and brain magnetic resonance imaging changes in subgroups based on type of presenting event, baseline brain magnetic resonance imaging parameters, and demographic factors in the Controlled High-Risk Subjects Avonex Multiple Sclerosis Prevention Study (CHAMPS) trial. After the onset of a first demyelinating event, 383 patients with brain magnetic resonance imaging evidence of subclinical demyelination were treated with corticosteroids and randomly assigned to receive weekly intramuscular injections of 30 microg interferon beta-1a or placebo. The treatment effect within subgroups was assessed in proportional hazards models both for the development of clinically definite multiple sclerosis and for a combined outcome of development of clinically definite multiple sclerosis or >1 new or enlarging T2 lesions on brain magnetic resonance imaging. A beneficial effect of treatment was noted in all subgroups evaluated. Adjusted rate ratios for the development of clinically definite multiple sclerosis in the optic neuritis, brainstem-cerebellar, and spinal cord syndrome subgroups were 0.58 (p = 0.05), 0.40 (p = 0.03), and 0.30 (p = 0.01) and for the development of the combined clinically definite multiple sclerosis/magnetic resonance imaging outcome were 0.50 (p < 0.001), 0.41 (p = 0.001), and 0.40 (p = 0.004), respectively. A treatment benefit on both outcome measures also was seen in subgroups based on baseline brain magnetic resonance imaging parameters, gender, and age. Interferon beta-1a is beneficial when initiated at the first clinical demyelinating event in patients with brain magnetic resonance imaging evidence of subclinical demyelination. The beneficial effect is present for optic neuritis, brainstem-cerebellar syndromes, and spinal cord syndromes. PMID- 11921055 TI - Subjective memory complaints: objective neural markers in patients with Alzheimer's disease and major depressive disorder. AB - Patients with probable Alzheimer's disease and depressive patients frequently present with subjective memory complaints. Objective distinction of underlying neuronal substrate malfunction and early cross-sectional differential diagnosis have been elusive thus far. We used repetitive learning and free recall of abstract geometric patterns during functional magnetic resonance imaging to assess episodic memory in older subjects (ages 56-64 years) who sought first-time medical attention with subjective memory complaints and were diagnosed with probable Alzheimer's disease (NINCDS-ADRDA criteria; ages 51-67 years) or major depressive disorder (DSM-IV; ages 50-65 years). Contrasting healthy seniors or depressive patients with Alzheimer's disease patients revealed superiority of hippocampal activation. Contrasting Alzheimer's disease patients with seniors showed bilateral prefrontal activity as a correlate of futile compensation of episodic memory failure. Contrasting patients who had major depressive disorder with seniors or patients who had Alzheimer's disease showed bilateral activation of the orbitofrontal cortex and the anterior cingulate. Subjective memory complaints may be classified objectively and very early with functional magnetic resonance imaging of episodic memory in groups of patients with Alzheimer's disease and depressive syndrome. This may facilitate drug trials with evaluation of specific treatments, but further studies will be needed to establish the differential diagnosis for the individual patient. PMID- 11921056 TI - Cortical spreading depression and gene regulation: relevance to migraine. AB - Cortical spreading depression (CSD) may be the underlying mechanism of migraine aura. The role of CSD in initiating a migraine headache remains to be determined, but it might involve specific changes in gene expression in the brain. To examine these changes, four episodes of CSD at 5-minute intervals were induced in the mouse brain by application of 300mM KCl, and gene expression was examined 2 hours later using cDNA array and reverse transcriptase-polymerase chain reaction. Controls consisted of groups that received anesthesia only, attachment of recording electrodes only, and application of 0.9% NaCl. Of the over 1,180 genes examined in our experiments, those consistently regulated by CSD included vasoactive peptides; the vasodilator atrial natriuretic peptide was induced by CSD, while the vasoconstrictor neuropeptide Y was downregulated. Other genes specifically regulated by CSD were involved in oxidative stress responses (major prion protein, glutathione-S-transferase-5, and apolipoprotein E). L-type calcium channel mRNA was upregulated. In summary, CSD regulates genes that are intrinsic to its propagation, that identify accompanying vascular responses as a potential source of pain, and that protect against its potential pathological consequences. We believe these observations have strong relevance to the mechanisms of migraine and its outcomes. PMID- 11921058 TI - Cell death in experimental intracerebral hemorrhage: the "black hole" model of hemorrhagic damage. AB - Intracerebral hemorrhage (ICH) has a poor prognosis that may be the consequence of the hematoma's effect on adjacent and remote brain regions. Little is known about the mechanism, location, and severity of such effects. In this study, rats subjected to intracerebral blood injection were examined at 100 days. Stereology (neuronal count and density) and volume measures in the perihematoma rim, the adjacent and overlying brain, and the substantia nigra pars reticulata (SNr) were compared with contralateral brain regions at 100 days and the perihemorrhage region at 24 hours and 7 days. In addition, cytochrome c release was investigated at 24 hours, 3 days, and 7 days. At 100 days, post-ICH rats showed no difference in neuronal density in the perihemorrhagic scar region or regions of the striatum immediately surrounding and distal to the perihemorrhage scar. The cell density index in the ipsilateral field was 16.2 +/- 3.8 versus the contralateral control field of 15.6 +/- 3.2 (not significant). Volume measurements of the ipsilateral striatum revealed a 20% decrease that was compensated by an increase in ipsilateral ventricular size. The area of the initial ICH as measured by magnetic resonance imaging correlated with the degree of atrophy. In the region immediately surrounding the hematoma, cytochrome c immunoreactivity increased at 24 hours and 3 days, and returned toward baseline by day 7. At 24 hours, stereology in the peri-ICH region showed decreased density in the region where cytochrome c immunoreactivity was the highest. Neuronal density of the ipsilateral SNr was significantly less than the contralateral side (9.6 +/- 1.9 vs 11.6 +/- 2.3). Histologic damage from ICH occurred mainly in the immediate perihemorrhage region. Except for SNr, we found no evidence of neuronal loss in distal regions. We have termed this continued destruction of neurons, which occurs over at least 3 days as the neurons come into proximity to the hematoma, the "black hole" model of hemorrhagic damage. PMID- 11921057 TI - Nuclear factor-kappaB as a molecular target for migraine therapy. AB - Nitric oxide (NO) generated from inducible NO synthase (iNOS) participates in immune and inflammatory responses in many tissues. The NO donor glyceryl trinitrate (GTN) provokes delayed migraine attacks when infused into migraineurs and also causes iNOS expression and delayed inflammation within rodent dura mater. Sodium nitroprusside, an NO donor as well, also increases iNOS expression. Because inflammation and iNOS are potential therapeutic targets, we examined transcriptional regulation of iNOS following GTN infusion and the consequences of its inhibition within dura mater. We show that intravenous GTN increases NO production within macrophages. L-N(6)-(1-iminoethyl)lysine, a selective iNOS inhibitor, attenuates the NO signal, emphasizing the importance of enzymatic activity to delayed NO production. iNOS expression is preceded by significant nuclear factor kappa B (NF-kappaB) activity, as reflected by a reduction in the inhibitory protein-kappa-Balpha (IkappaBalpha) and activation of NF-kappaB after GTN infusion. IkappaBalpha degradation, NF-kappaB activation, and iNOS expression were attenuated by parthenolide (3mg/kg), the active constituent of feverfew, an anti-inflammatory drug used for migraine treatment. These findings suggest that GTN promotes NF-kappaB activity and inflammation with a time course consistent with migraine attacks in susceptible individuals. We conclude, based on results with this animal model, that blockade of NF-kappaB activity provides a novel transcriptional target for the development of anti-migraine drugs. PMID- 11921059 TI - Late-onset frontotemporal dementia with a novel exon 1 (Arg5His) tau gene mutation. AB - We report a case of frontotemporal dementia and parkinsonism linked to chromosome 17 of 5 years' duration in an 81-year-old man whose brother had died at age 86 years with dementia. In this patient, we found frontal and temporal neuronal loss, glial-predominant tau deposits, progressive supranuclear palsy-like straight tubules, accumulation of 4-repeat-predominant Sarkosyl-insoluble tau, and a novel exon 1 (Arg5His) tau gene mutation. This mutation decreased microtubule-promoting capacity and increased fibrillation of tau in vitro. Thus, we consider that the Arg5His mutation is an authentic tau gene abnormality responsible for the patient's tau pathology and late-onset dementia. PMID- 11921060 TI - Levodopa-induced dyskinesias in normal squirrel monkeys. PMID- 11921062 TI - Myelin-derived glycolipids and animal models of Guillain-Barre syndrome. PMID- 11921064 TI - New diagnostic criteria for multiple sclerosis. PMID- 11921069 TI - Developing a model of colchicine neuropathy. AB - Colchicine may be a cause of neuropathy. An experimental model to study this is developed. PMID- 11921070 TI - Beneficial effect of hyperbaric oxygen on island flaps subjected to secondary venous ischemia. AB - The potential for hyperbaric oxygen therapy (HBO) to decrease the untoward effects of a secondary ischemic event was studied in the rat superficial epigastric flap model. The secondary venous ischemic flap was created by cross clamping the vascular pedicles for 2 h. Twenty-four hours later, the flap was reelevated and the venous pedicle was occluded for 5 h. Thirty-two rats were divided into three groups. In experimental group 1, animals received HBO treatment immediately prior to the initial flap elevation and ischemia at 2 atmosphere pressures for 90 min. In experimental group 2, the rats underwent a similar course except for a second 90-min HBO course immediately prior to the secondary venous occlusion. The rats without HBO therapy were used as controls. The results showed that all control flaps were nonviable at 1 week by clinical inspection and fluorescein injection. Complete flap survival occurred in 20% of group 1 flaps and 30.8% of group 2 flaps. Partial flap survival occurred in the rest of the flaps in these two groups, with mean survival areas of 48% and 55%, respectively. In conclusion, HBO treatments significantly increase the survival of flaps subjected to a secondary ischemia, even if administered before the primary ischemia. Administering HBO prior to secondary venous ischemia was marginal, which may be due to the effect of O(2) given by HBO not lasting longer than 5 h. PMID- 11921071 TI - Nasal columella reconstruction with a composite free flap from the root of auricular helix. AB - Defects of the nasal columella result in significant cosmetic and functional deformities. Over the years, a variety of methods for nasal columella reconstruction have been described in the literature, as have the technical difficulties of reconstructing this subtle structure. Here, a successful reconstruction of a 3.0 x 2.0 cm-wide nasal columella defect, with a chondrocutaneous microsurgical free flap from the root of the auricular helix, is presented. PMID- 11921072 TI - Secondary surgery after digit replantation: its incidence and sequence. AB - This article discusses the incidence and sequence of secondary procedures following digital replantation. In published series addressing this topic, the overall frequency of postreplantation surgery ranged from 2.9-93.2%. Tendon procedures accounted for 47.2% of cases, and comprised the leading type of secondary operations. Tendon procedures were followed by joint procedures (18.9%), skeletal stabilization (12%), skin coverage (11.4%), nerve reconstruction (8.9%), and late amputation (1.6%). The number of secondary procedures per patient averaged from 1-4.5, depending on prioritization of different procedures. The order of restoration procedures should be as follows: supple skin coverage, skeletal stability, and protective and proprioceptive sensation. Joint reconstruction follows these procedures, which in turn is followed by tendon reconstruction. PMID- 11921073 TI - Modified sleeve anastomosis for reconstruction of the hepatic artery in rat liver transplantation. AB - End-to-end sleeve anastomosis between a donor common hepatic artery and a recipient proper hepatic artery was proven to be the most physiological and simple method for hepatic rearterialization in rat liver transplantation. Current technical variants of the sleeve technique, however, are hampered by the high rate of bleeding from the anastomotic site. This report deals with a technical modification which inhibits postoperative bleeding efficiently. The procedure consisted of a guiding suture, as previously described in other technical variants, and a modified fixing suture. Instead of using a single stitch to fix the feeding vessel with the receiving vessel, a running suture between the edge of the donor common hepatic artery and the adventitia of the recipient proper hepatic artery was performed to avoid a possible backflow. The patency rate of 91% was as high as reported by others using a sleeve technique, which was also reflected in the histomorphological picture, being indistinguishable from normal liver histology. This technical modification simplified the procedure of reconstructing the hepatic artery and could contribute to a wider use of the arterialized liver transplantation model in rats. PMID- 11921074 TI - Effects of University of Wisconsin and lactated Ringer's solutions to ischemia reperfusion injury in isolated cremaster flap. AB - Ischemia-reperfusion (I/R) injury is a topic that has been much-discussed by various researchers during the last decade in plastic surgery. Though much progress has occurred, the problem is not totally solved yet. In particular, the pathophysiology of reperfusion injury in skeletal muscle has not been clearly elucidated. The aims of this study are to assess the effects of a variety of perfusants on the microcirculation after reperfusion injury and to better understand the pathophysiology of reperfusion injury. Isolated cremaster flaps were performed in 44 rats, preserving the femoral artery and vein in order to cannulate with microtubes. There were 2 control and 2 experiment groups. In one of the control groups and in both experimental groups, 2 h of ischemia were applied by clamping the iliac vessels. Immediately after this, the muscle was locally perfused and washed with lactated Ringer's (LR) and University of Wisconsin (UW) solutions, given from the femoral artery and drained by the femoral vein in the two respective experimental groups. The effects of these solutions to I/R injury were shown at the microcirculatory level via measuring and determining preischemic and postischemic diameters of arterioles and venules, tissue perfusion, capillary density, velocity of red blood cells, and leukocyte sticking. Both tested perfusion solutions were found to be harmful in all parameters. This study demonstrates that both LR and UW solutions aggravate I/R injury. PMID- 11921075 TI - Lymphedema and microsurgery. AB - Lymphedema is often diagnosed by its characteristic clinical presentation. In some cases, however, instrumental investigations are necessary to establish the diagnosis, particularly in early stages of the disease. One of the primary problems for microsurgery in treating lymphedema consists of the discrepancy between the excellent technical possibilities and the insufficient results in reduction of lymphedematous tissue fibrosis and sclerosis. Long-term results indicate that microsurgical operations have a valuable place in the treatment of obstructive lymphedema (primary or secondary) and should be the treatment of choice in these patients. Improved results can be expected with earlier microsurgical operations because patients referred earlier usually have less lymphatic disruption and fibrotic tissue. Advanced diagnostic methods and improvements in operation techniques have modified indications for surgical therapy of lymphedema. This article systematically reviews the published literature on the microsurgical treatment of lymphedema to the present. PMID- 11921076 TI - Effects of hypocholesterolaemic agents on the expression and activity of 3 hydroxy-3-methylglutaryl-CoA reductase in the fat body of the German cockroach. AB - In the fat body of adult Blattella germanica females, the expression of 3-hydroxy 3-methylglutaryl-CoA reductase (HMG-CoA reductase) during the first reproductive cycle is parallel to that of vitellogenin, suggesting a functional link between the mevalonate pathway, and vitellogenesis and reproduction. We have studied the effects of compactin and fluvastatin, two inhibitors of HMG-CoA reductase, on the expression and activity of the enzyme in the fat body, and on the ootheca formation, ootheca viability, and number of larvae per viable ootheca. Short-term assays showed that both compounds reduce the protein levels and enzymatic activity of HMG-CoA reductase, and long-term experiments revealed that fluvastatin impairs embryo development. PMID- 11921077 TI - Influence of calcium on Manduca sexta plasmatocyte spreading and network formation. AB - Plasmatocytes are a class of insect hemocytes important in the cellular defense response. In some species, they are phagocytic, protecting the insect from smaller pathogens. In many insects, they work in concert with other hemocytes (particularly other plasmatocytes and granular cells) to form nodules and to encapsulate foreign material. To perform these functions, plasmatocytes attach to, spread on, and surround suitable targets. Because of their importance, because we had previously observed that prolonged incubation of hemocytes in solutions containing the divalent cation chelator ethylenediaminetetraacetic acid (EDTA) inhibited plasmatocyte spreading, and because of the importance of divalent cations in many immune-related functions, we investigated the effect of calcium and magnesium on spreading of plasmatocytes from fifth instar Manduca sexta larvae. On glass slides, plasmatocytes spread more quickly and elongated in Grace's medium containing 5 mM calcium, compared to calcium-free medium. In the presence of calcium, plasmatocyte adhesion, spreading, and network formation were not visibly different in magnesium-free and magnesium-containing Grace's medium. Using immunomicroscopy with a monoclonal antibody specific for plasmatocytes, we measured the length and width of plasmatocytes incubated with several different concentrations of calcium. Plasmatocyte length positively correlated with calcium concentration to 5 mM (maximum concentration tested and approximately the hemolymph concentration). Mean plasmatocyte width was less in 0 and 5 mM calcium than in 0.05 or 0.5 mM calcium. On plastic, hemocytes survived longer than on glass (they survived beyond 24 h) and, in 5 mM calcium, formed an extensive network readily visible by phase-contrast microscopy. This network was never as extensive in the absence of calcium. Network formation in the absence of magnesium, but presence of calcium, resembled network formation in standard Grace's medium. PMID- 11921078 TI - Effect of geminivirus infection and Bemisia infestation on accumulation of pathogenesis-related proteins in tomato. AB - The whitefly, Bemisia tabaci biotype B, has been shown to cause pathogenesis related (PR) proteins to accumulate in plants as a result of direct feeding, but their specific role in plant defensive systems is unclear. Our objective was to compare accumulation of tomato PR proteins (beta-1,3-glucanase, chitinase, peroxidase, P2 and P4) in response to whitefly, with or without tomato mottle virus (ToMoV) infection. Tomato PR protein response was measured over time in plants divided into three treatments: uninfected controls (with or without whiteflies) and plants infested with viruliferous (ToMoV) whiteflies. Five- to six-leaf plants were infested with approximately 5 adult whitefly per leaf. Plants were sampled prior to whitefly infestation and at 14, 28, 42, and 56 days. By 56 days, plants infested with viruliferous whiteflies had significantly more eggs (2.5-fold) and nymphs (4.5-fold) than plants with nonviruliferous whiteflies. A significant increase in the enzymatic activity of all measured PR proteins, as compared to control plants, was only seen in viruliferous whitefly infested plants. No significant difference was observed in enzyme activities between the uninfected control plants either with or without whiteflies. The greatest differences for all PR proteins assayed were observed 42 days after treatment initiation. Protein blot analyses showed that the differences in PR protein activities among the treatments were due to changes in specific enzyme levels within the plant and were associated with concomitant increases in levels of P2 and P4 PR proteins. Under our experimental conditions, it is clear that PR protein response is much more intense when it is attacked by whiteflies carrying ToMoV than by whitefly alone. PMID- 11921079 TI - Cholinergic regulation of the corpora allata in adult male loreyi leafworm Mythimna loreyi. AB - The direct effect of acetylcholine on the activation of the corpora allata (CA) was investigated in the adult male loreyi leafworm, Mythimna loreyi. Acetylcholine, in the presence of the choline esterase inhibitor physostigmine (50 microM), elicited a stimulatory effect on juvenile hormone acids (JHAs) release from the CA. Maximum effect was obtained at concentrations of 10 and 50 microM. Repeated administration of 10 microM acetylcholine on the same CA did not elicit similar stimulatory effect. Since JHA release can be significantly activated by carbachol and not by nicotine, this cholinergic effect is likely to belong to the muscarinic type. The effect of acetylcholine was significantly antagonized by gallamine triethiodide (M(2) antagonist) and 4-DAMP (M(3) antagonist), pirenzepine (M(1) antagonist), and tropicamide (M(4) antagonist) were ineffective. It is concluded that in the adult male M. loreyi, the cholinergic regulation of CA is most likely via M(2) and M(3) muscarinic receptors. PMID- 11921081 TI - Characteristics of genetic markers and maps for cost-effective genome screens using diallelic markers. AB - New technologies for genotyping diallelic markers (SNPs) were recently developed that may be lower in cost, and more easily automated than microsatellite markers (STRPs). The reduction in genotyping costs resulting from such automation may significantly impact the overall cost of studies of complex traits, which generally require large sample sizes. Use of multiple SNPs in linkage analysis can recapture the linkage information otherwise lost with such markers. Here we derive a measure of the multilocus polymorphic information content (MPIC) in the context of linkage analysis for a cluster of SNPs, and we explore the characteristics of uniform vs. clustered SNP maps, relative to STRP maps. Issues addressed in comparing the map structures include the information content for clustered or single markers, and map accuracy. To be cost-effective, SNPs should have a common allele frequency between 0.5-0.75. No more than five loci per cluster are needed. Some linkage disequilibrium between loci in a cluster is tolerable. In the ideal case, a uniformly spaced SNP map is more cost-effective than one composed of clustered loci. However, the genotyping cost per marker for diallelic markers can be at most 60% of the genotyping cost per marker for STRPs. The consequences of using clustered vs. uniform SNP maps are considered in the context of map inaccuracy and use of multipoint vs. pairwise linkage analysis. Overall, when marker information, map accuracy, and flexibility of analysis are jointly considered, an optimal solution may be use of maps with 2-3 SNPs per cluster. PMID- 11921082 TI - Extension of variance components approach to incorporate temporal trends and longitudinal pedigree data analysis. AB - Here we present a method that permits one to evaluate genetic effects and to detect genetic linkages by using serial observations of quantitative traits in pedigrees. We developed a statistical method that incorporates longitudinal family data and genetic marker information into an estimating equations framework. With this approach, we can study changes in components over time that measure polygenic and major genetic variances as well as shared and individual specific environmental effects. Our method provides a measure of heritability from analysis of longitudinal data. Results using longitudinal family data from the Center for Preventive Medicine (Nancy, France) are presented. The results of our analysis show that the apolipoprotein E locus has no effect on interindividual variability in systolic blood pressure. We found that the longitudinal measure of heritability of systolic blood pressure is 0.32. PMID- 11921083 TI - LPL polymorphism predicts stroke risk in men. AB - Variation in lipid levels has been associated with atherosclerotic vascular disease, including stroke. Genes contributing to interindividual variation in lipid levels may play a role in the etiology of stroke, either through their effects on lipid synthesis and metabolism or through separate pathways. For this reason, we sought to examine the association between polymorphisms in the lipoprotein lipase (LPL) and apolipoprotein E (APOE) genes and subclinical and clinical stroke in the Atherosclerosis Risk in Communities (ARIC) Study. Subclinical stroke was determined by cerebral magnetic resonance imaging (MRI). Subclinical cerebral infarct cases (n = 197) were compared to a stratified random sample identified from individuals participating in the MRI examination (n = 200). Incidence of clinical ischemic stroke was determined by following the ARIC cohort for an average of 7.5 years for potential cerebrovascular events; 218 validated clinical ischemic strokes were identified. A stratified random sample of the ARIC cohort (CRS, n = 964) was used as the comparison group for clinical cases. The LPL S291-carrying genotypes and APOE epsilon2- and epsilon4-carrying genotypes were not significantly associated with subclinical or clinical stroke. The LPL X447-containing genotypes were significantly associated with subclinical (odds ratio [OR], 4.32; 95% confidence interval [CI], 1.23-15.15; P = 0.020) and clinical stroke (hazard rate ratio [HRR], 2.57; 95% CI, 1.24-5.34; P = 0.01) in men, both by themselves and after adjustment for multiple stroke risk factors. The LPL S447X polymorphism is significantly associated with subclinical cerebral infarction and incident clinical ischemic stroke in men from a middle-aged American population. This association does not appear to be mediated by triglyceride, high-density lipoprotein (HDL)- and low-density lipoprotein (LDL) cholesterol levels, or additional stroke risk factors. PMID- 11921084 TI - Genetics of the apnea hypopnea index in Caucasians and African Americans: I. Segregation analysis. AB - Differences in age of presentation and anatomic risk factors for obstructive sleep apnea (OSA) in Caucasians and African Americans suggest possible racial differences in the genetic underpinnings of the disorder. In this study, we assess transmission patterns in a Caucasian sample consisting of 177 families (N = 1,195) and in an African American sample consisting of 125 families (N = 720) for two variables: 1) apnea hypopnea index (AHI) log transformed and adjusted for age, and 2) AHI log transformed and adjusted for age and body mass index (BMI). We allowed for residual familial correlations and sex-specific means in all models. Analysis of the Caucasian sample showed transmission patterns consistent with that of a major gene that were stronger in the age-adjusted variable than in the age- and BMI-adjusted variable. However, in the African American families, adjusting for BMI in addition to age gave stronger evidence for segregation of a codominant gene with an allele frequency of 0.14, accounting for 35% of the total variance. These results provide support for an underlying genetic basis for OSA that in African Americans is independent of the contribution of BMI. PMID- 11921085 TI - Genetic component to susceptibility to Trichuris trichiura: evidence from two Asian populations. AB - Trichuris trichiura is an helminthic infection with potentially severe health consequences. The hypothesis that host genetic factors can account for the distribution of Trichuris was tested using familial data on egg counts available for two populations, the Jirels of Nepal and the population of Jishan Island in Jiangxi Province of the People's Republic of China. Whipworm is highly prevalent in the Jishan Island population (86%), but occurs at a low rate in the Jirel population (14%). A quantitative genetic analysis was performed on each data set, using a variance component approach. Approximately 28% of the variation in Trichuris trichiura loads was attributable to genetic factors in both populations. Common household effects accounted for only 4% of the variation in the Jirels and none of the variation in the Jishan Island population. These concordant results from two separate populations provide strong evidence of the important role of genetics in determining differential susceptibility to whipworm infection. PMID- 11921086 TI - Multiple sclerosis recurrence risk for siblings in an isolated population of Central Sardinia, Italy. AB - Studies of twins, adoptees, half siblings, and familial recurrence risk have shown that genetic and non-genetic factors are involved in multiple sclerosis (MS) etiology. Age at onset, gender, and parental MS status seem to influence sibling risk. We studied the recurrence risk in siblings of MS patients in an isolated population of Sardinia, Italy, which is genetically homogeneous, inbred, and very stable, with a high MS frequency. The Aalen-Nelson estimate of the recurrence risk in siblings is 4.7%, and the risk ratio compared with the general population is 31. Proportional hazards models were used to investigate the effect of sibling sex, sex, and age at onset of the proband, and number of affected relatives on a sibling's predicted MS risk. Sib's risk is influenced by age at onset (P = 0.02), and possibly by sex of the proband (P = 0.08). There is also a borderline significant interaction (P = 0.05) between the sex and age at onset of the proband: early age at onset influences sib's risk only if the proband is female. The number of affected relatives in the family is not found to influence sibling risk, but the power is lacking (95% CI 0.50-2.62). This result is consistent with a single dominant gene with an extremely low penetrance, a model that has not yet been disproved as a possible inheritance model for MS. PMID- 11921087 TI - The role of genotype in determining the effects of cigarette smoking on pulmonary function. AB - The effect of cigarette smoking on pulmonary function is highly variable. Some heavy smokers retain normal pulmonary function and others experience profound pulmonary function loss. The role of genotype in this process is unknown. We tested for gene by environment interaction (GxE) in smoking-associated loss of forced expiratory volume in one second (FEV1) using repeated pulmonary function and smoke-exposure measures from 352 twin pairs enrolled in the NHLBI Male Veteran Twin Study. Inferences using global [Jinks and Fulker, 1970] and dichotomous [Ramakrishnan et al., 1992] procedures were supplemented with a new model for repeatedly observed twinships. The model facilitates testing of standard heritability hypotheses and families of hypotheses regarding the dependence of quantitative twin-twin phenotypic similarity on continuously varying twin-twin exposure concordance. The global and dichotomous procedures were suggestive of GxE in smoke-associated loss of FEV1 (p < 0.01, p = 0.08, respectively). With the new model, overall twin-twin correlation of FEV1 for concordant-smoking MZ and DZ twin pairs was estimated at 0.71 and 0.34, respectively. For twins with little or no difference in cigarette use, the intra pair correlations of FEV(1) did not differ according to cigarette exposure over a wide range of exposures (0 - > or = 200 pack years). Even great twin-twin discordance in cigarette smoking (> or =10 pack years) had little effect of correlations. We conclude that a constant factor, such as genotype, appears to be interposed between the environmental toxin (cigarette smoke) and phenotype (FEV1). PMID- 11921088 TI - Sets of integrating plasmids and gene disruption cassettes containing improved counter-selection markers designed for repeated use in budding yeast. AB - Counter-selection is a useful gene manipulation technique for repeated gene disruptions, gene shufflings and gene replacements in yeasts. We developed a novel counter-selection system using a galactose-inducible growth inhibitory sequence (Kawahata et al.1999. Yeast 15: 1-10). This counter-selection marker, named GAL10p-GIN11, has several advantages over previous counter-selection markers, i.e. use of an inexpensive galactose medium for counter-selection, combined use with any transformation markers for gene introduction, and no requirement of specific mutations in the host strains. The GIN11 sequence, which is a part of an X-element of the subtelomeric regions, contained a conserved autonomously replicating sequence, causing the possibility of inefficient chromosomal integration. We isolated GIN11 mutants that lost the replication activity but retained the growth-inhibitory effect when overexpressed. A mutant GIN11M86 sequence was selected and fused to the CUP1 promoter for the counter selection on a copper-containing medium. The GALp-GIN11M86 and the CUPp-GIN11M86 were used for constructing sets of integrating plasmids containing auxotrophic markers involving HIS3, TRP1, LEU2, URA3 or ADE2, or a drug-resistant marker PGKp YAP1. In addition, a set of gene disruption cassettes that contained each of the auxotrophic markers and the GALp-GIN11M86, which were flanked by direct repeats of a hisG sequence, were constructed. The counter-selectable integrating plasmids and the gene disruption cassettes can allow the markers to be used repeatedly for yeast gene manipulations. PMID- 11921089 TI - Systematic analysis of sporulation phenotypes in 624 non-lethal homozygous deletion strains of Saccharomyces cerevisiae. AB - A new high throughput mutant screening procedure for the detection of sporulation mutants was developed and used to analyse a set of 624 non-lethal homozygous deletion mutants created in the European joint research program EUROFAN. The screening procedure involved determination of LL- and DL-dityrosine, sporulation specific compounds, which were shown to be robust markers of the extent and arrest stage of sporulation mutants. Secondary screens consisted of light microscopy to detect mature and immature spores and DAPI staining to monitor the progress of meiotic nuclear divisions. We discovered new phenotypic classes of mutants defective in spore wall synthesis that were not discovered by previous screens for sporulation mutants. The genes corresponding to the sporulation mutants fell in several functional classes, some of which were previously unknown to be involved in spore formation. Peroxisomes seem to play a role in spore wall synthesis. Mitochondria play a role in sporulation that is not simply restricted to supply of ATP from respiratory metabolism. The deletion mutants included in the set were functionally unknown at the start of EUROFAN; however, within the last few years the importance to sporulation of some of them was also reported by other authors. Taken together, about 8% of all single gene deletion mutants of non-essential genes of Saccharomyces cerevisiae seem to display a clear and reproducible sporulation phenotype. PMID- 11921090 TI - Functional characterization of the Candida albicans homologue of secretion associated and Ras-related (Sar1) protein. AB - Secretion-associated and Ras-related protein (Sar1p) plays an essential role during the protein transport from the endoplasmic reticulum to the Golgi apparatus. The cDNA sequence of the Sar1 gene has been identified and characterized from the human yeast pathogen, Candida albicans. This cDNA encodes a protein of 190 amino acids, which shares a 78% sequence identity with Saccharomyces cerevisiae Sar1p and contains the conserved GTP-binding motifs of the small GTPase superfamily. Complementation studies confirmed that this cDNA encodes the functional homologue of ScSar1p. The recombinant C. albicans Sar1p exhibits GTP-binding activity in vitro that was abolished by deletion of one of the three GTP-binding motifs. PMID- 11921091 TI - Saccharomyces cerevisiae and Zygosaccharomyces mellis exhibit different hyperosmotic shock responses. AB - The effect of hyperosmotic shock on cell volume, vacuole volume, and intracellular pH (pH(i)) of individual cells of Saccharomyces cerevisiae and Zygosaccharomyces mellis was investigated. After transfer from a high water activity (a(w)) medium to low a(w) media, the growth latency periods of Z. mellis were shorter than those of S. cerevisiae. These results demonstrate that Z. mellis manages hyperosmotic shock better than S. cerevisiae. As a response to acute hyperosmotic shock, i.e. the first minute of perfusion with hypertonic buffers, the vacuoles shrank and the pH(i) decreased in both yeasts. Furthermore, in the presence of glucose, vacuole shrinkage and intracellular acidification were more pronounced in S. cerevisiae than in Z. mellis. These results may be explained by the fact that the S. cerevisiae cells shrank more than the Z. mellis cells as a response to acute hyperosmotic shock. In the presence of glucose, the vacuoles and the cells of both S. cerevisiae and Z. mellis shrank simultaneously and in proportion to a minimum level during acute hyperosmotic shock, and remained constant at this level throughout the experiment (11 min). These results indicate that vacuoles do not act as water reserves in yeasts after acute hyperosmotic shock. Finally, Z. mellis was able to maintain its pH(i) near normal physiological levels after acute hyperosmotic shock, whereas S. cerevisiae was not. These results suggest that pH(i) regulation may be important for the ability of yeasts to manage hyperosmotic shock. PMID- 11921092 TI - Tools for the study of genome rearrangements in laboratory and industrial yeast strains. AB - In order to investigate the extent of genome rearrangements in laboratory and industrial yeast strains, a set of plasmids, containing ca. 300 bp fragments from highly conserved genes from S. cerevisiae, has been constructed. We chose three unique PCR products, each from a single gene, per chromosome: one from close to the centromere, and one from each chromosome end. Using these plasmids as probes to hybridize a Southern blot from a pulsed-field gel electrophoresis separation of the 16 yeast chromosomes, it is possible to identify large chromosomal rearrangements such as reciprocal translocations. PMID- 11921093 TI - Enzymatic activities of Ura2 and Ura1 proteins (aspartate carbamoyltransferase and dihydro-orotate dehydrogenase) are present in both isolated membranes and cytoplasm of Saccharomyces cerevisiae. AB - Computational analysis predicted three potential hydrophobic transmembrane alpha helices within the Ura2 multidomain protein of Saccharomyces cerevisiae, the C terminal subdomain of which catalyses the second step of uridine-monophosphate biosynthesis by its L-aspartate carbamoyltransferase activity (EC 2.1.3.2). The fourth step of pyrimidine biosynthesis is catalysed by dihydro-orotate dehydrogenase (Ura1 protein; EC 1.3.99.11), which was similarly characterized as a peripheral membrane protein. Ex situ, the activities of the investigated enzymes were associated both with isolated yeast membranes, fractionated by differential centrifugation to remove intact nuclei, and with soluble cytoplasmic proteins. PMID- 11921095 TI - Bibliography. Current awareness on yeast. PMID- 11921094 TI - Molecular and functional analysis of a MIG1 homologue from the yeast Schwanniomyces occidentalis. AB - A putative glucose repressor MIG1-homologue (SoMIG1) was isolated from the amylolytic yeast Schwanniomyces occidentalis. Degenerate primers were designed from the conserved zinc finger regions of Mig1 and CreA proteins from different organisms. PCR using these primers and S. occidentalis genomic DNA as template yielded a single 128 bp product. This fragment was used as a DNA probe to screen a S. occidentalis genomic library. Analysis of the positive clones led to the isolation by PCR of a DNA fragment, which contained an open reading frame (ORF) that would encode a 458 amino acid polypeptide. The DNA binding and effector domains of this putative protein showed an identity of 71% and 15%, respectively, to those of the Mig1 protein from Saccharomyces cerevisiae. The SoMIG1 gene complemented a mig1 mutant of this yeast, which suggests that in S. occidentalis SoMIG1 is a glucose repressor. The Accession No. is AJ417892. PMID- 11921096 TI - Increasing sulphite formation in Saccharomyces cerevisiae by overexpression of MET14 and SSU1. AB - Saccharomyces cerevisiae produces sulphite as an intermediate product during the assimilatory reduction of sulphate to sulphide. Three genes, MET3, MET14 and MET16, are essential for this reduction. We investigated the level of transcription of these genes in strains of S. cerevisiae with high, medium and low sulphite formation. The level of MET14- and MET16-mRNA varied with sulphite production, whereas the level of MET3-mRNA was very weak in almost all strains. We also analysed the effect of overexpression of MET14 and MET16 on sulphite formation. Two strains with low sulphite production were transformed with high copy plasmids containing either or both MET14 and MET16. The overexpression of these two genes leads to a two- to three-fold sulphite formation. In addition, inactivation of MET10, encoding a subunit of the sulphite reductase, also leads to a distinct increase in sulphite formation; however, the cells became methionine auxotroph. The overexpression of SSU1, a gene encoding a putative sulphite pump, yields a slight increase in sulphite accumulation, whereas overexpression of SSU1, together with MET14, increases sulphite formation up to 10-fold. Furthermore, sulphite formation strongly depends on growth conditions, e.g. yeast transformants growing in wort produce much higher amounts of sulphite when compared to growth in minimal media. The addition of glucose can also increase the sulphite formation in strains overexpressing MET14 and/or SSU1 under oxygen-limiting conditions, while the addition of glucose has no significant effect under aerobic conditions. PMID- 11921097 TI - Marker construction and cloning of a cut1-like sequence with ARS activity in the fission yeast Schizosaccharomyces japonicus. AB - The dimorphic fission yeast Schizosaccharomyces japonicus has proved to be an excellent experimental model for the investigation of the eukaryotic cell. Here we show that it has a haplontic life cycle, in which the diploid phase is confined to the zygote. To make it amenable to genetic and molecular analysis, we generated genetic markers and cloned a genomic sequence which acts as ars when integrated into a plasmid. Diploids suitable for testing complementation and recombination between markers can be formed by protoplast fusion. The complementation tests and the recombination frequencies determined in octads of spores identified 28 non-allelic groups (genes) of mutations of the auxotrophic and mycelium-negative mutants. Two groups of linked markers were also identified. The cloned fragment, which expresses ars activity, encodes a putative amino acid sequence highly similar to a conserved domain of proteins Cut1 (Schizosaccharomyces pombe), BimB (Aspergillus nidulans) and Esp1 (Saccharomyces cerevisiae). PMID- 11921098 TI - In vivo selectively infective phage as a tool to detect protein interactions: evaluation of a novel vector system with yeast Ste7p-Fus3p interacting proteins. AB - The selectively infective phage (SIP) approach allows rapid identification of interacting proteins by linking protein-protein interaction to phage infectivity. Infection of E. coli by filamentous phage depends on viral g3p. This protein consists of three domains, N1, N2 and CT. Phages lacking the N1 domain are non infective unless a bait (X)-prey (Y) interaction links it to phage anchored N2-CT domains. We have developed all the vectors required for an in vivo selectively infective phage strategy (SIP). This includes a bait vector, pG3N1, a prey vector, pHOS41, and a gene III deletion helper phage, HPd3. The bait vector pG3N1 allows expression of a bait protein (X) fused to the C-terminus of the N1 domain. The prey vector pHOS41 allows expression of prey (Y) proteins, fused to the N terminus of the N2-CT domains. The gene III deletion helper phage delivers all phage proteins necessary for phage production, except g3p. Escherichia coli transformed with these three vectors produces non-infective phages unless a bait prey interaction links the g3p domains. Fus3p and Ste7p, two proteins from the Saccharomyces cerevisiae pheromone-responsive pathway have been cloned to evaluate the SIP strategy. The presence of the interacting N1-Fus3p adapter increased the infectivity of Ste7p-N2-CT phages approximately 1400-fold, which makes SIP a promising technology for the detection and further investigation of interacting proteins. PMID- 11921099 TI - Functional analysis of structural genes for NAD(+)-dependent formate dehydrogenase in Saccharomyces cerevisiae. AB - Co-consumption of formate by aerobic, glucose-limited chemostat cultures of Saccharomyces cerevisiae CEN.PK 113-7D led to an increased biomass yield relative to cultures grown on glucose as the sole carbon and energy substrate. In this respect, this strain differed from two previously investigated S. cerevisiae strains, in which formate oxidation did not lead to an increased biomass yield on glucose. Enzyme assays confirmed the presence of a formate-inducible, cytosolic and NAD(+)-dependent formate dehydrogenase. To investigate whether this enzyme activity was entirely encoded by the previously reported FDH1 gene, an fdh1Delta null mutant was constructed. This mutant strain still contained formate dehydrogenase activity and remained capable of co-consumption of formate. The formate dehydrogenase activity in the mutant was demonstrated to be encoded by a second structural gene for formate dehydrogenase (FDH2) in S. cerevisiae CEN.PK 113-7D. FDH2 was highly homologous to FDH1 and consisted of a fusion of two open reading frames (ORFs) (YPL275w and YPL276w) reported in the S. cerevisiae genome databases. Sequence analysis confirmed that, in the database genetic background, the presence of two single-nucleotide differences led to two truncated ORFs rather than the full-length FDH2 gene present in strain CEN.PK 113-7D. In the latter strain background an fdh1Deltafdh2Delta double mutant lacked formate dehydrogenase activity and was unable to co-consume formate. Absence of formate dehydrogenase activity did not affect growth on glucose as sole carbon source, but led to a reduced biomass yield on glucose-formate mixtures. These findings are consistent with a role of formate dehydrogenase in the detoxification of exogenous formate. PMID- 11921100 TI - The mei3 region of the Schizosaccharomyces pombe genome. AB - Expression of the mei3 gene is sufficient to induce meiosis in the fission yeast Schizosaccharomyces pombe. The mei3 gene is located 0.64 Mb from the telomere of the left arm of Sz. pombe chromosome II. We have sequenced and analysed 107 kb of DNA from the mei3 genomic region. The sequence includes 14 known genes (bag1-B, csh3, dps1, gpt1, mei3, mfm3, pac1, prp31, rpl38-1, rpn3, rti1, spa1, spm1 and ubc4) and 26 other open reading frames (ORFs) longer than 100 codons: a density of one protein-coding gene per 2.7 kb. Twenty-one of the 40 ORFs (53%) have introns. In addition there is one lone Tf1 transposon long terminal repeat (LTR), tRNA(Trp) and tRNA(Ser) genes and a 5S rRNA gene. 14 of the novel ORFs show sequence similarities which suggest functions of their products, including a coatomer alpha-subunit, a catechol O-methyltransferase, protein kinase, asparagine synthetase, zinc metalloprotease, acetyltransferase, phosphatidylinositol 4-kinase, inositol polyphosphate phosphatase, GTPase activating protein, permease, pre-mRNA splicing factor, 20S proteasome component and a thioredoxin-like protein. One predicted protein has similarity to the human Cockayne syndrome protein CSA and one with human GTPase XPA binding protein XAB1. Three ORFs are likely to code for proteins because they have sequence similarity with hypothetical proteins, three encode predicted coiled-coil proteins and four are sequence orphans. PMID- 11921101 TI - Insertional mutagenesis in yeasts using T-DNA from Agrobacterium tumefaciens. AB - Insertional mutagenesis is a powerful tool for the isolation of novel mutations. The gene delivery system of the bacterium Agrobacterium tumefaciens, which mediates transfer not only to plants but also to yeasts and fungi, could be exploited to generate collections of yeasts containing insertional mutations if there were no bias towards particular integration sites, as is the case in plants. To test this, we have analysed a small collection of Saccharomyces cerevisiae strains with T-DNA copies integrated in the S. cerevisiae genome. The position of 54 of these T-DNAs was determined. The T-DNA showed no clear preference for certain DNA sequences or genomic regions. We have isolated insertions in the coding regions of the genes YGR125w, YDR250c, YGR141w, YGR045c, YPL017c, YGR040w, YDL052c, YJL148w, YCL033c, YFL061w, YJR033c, YDR175c and YLR309c confirming that these genes are non-essential for S. cerevisiae haploid growth on minimal medium. Given the advantages of T-DNA, we propose its use as an ideal mobile DNA element for insertional mutagenesis in yeasts. PMID- 11921102 TI - A second Zn(II)(2)Cys(6) transcriptional factor encoded by the YNA2 gene is indispensable for the transcriptional activation of the genes involved in nitrate assimilation in the yeast Hansenula polymorpha. AB - Nitrate assimilation genes encoding a nitrate transporter (YNT1), nitrite reductase (YNI1), a Zn(II)(2)Cys(6) transcriptional factor involved in nitrate induction (YNA1) and the nitrate reductase (YNR1) are clustered in the yeast Hansenula polymorpha. A second gene, termed YNA2 (yeast nitrate assimilation), was located seven nucleotides away from the 3' region of YNR1 gene. The cluster is flanked by an ORF encoding a protein with similarity to glutathione-S transferase on the YNT1 side and an ORF with similarity to Saccharomyces cerevisiae Rad3p on the YNA2 side. The disruption of YNA2 confers the resulting null mutant strain with inability to grow in nitrate. The YNA2 gene encodes a putative protein of 618 residues bearing in the N-terminus the consensus sequence Cys-X(2)-Cys-X(6)-Cys-X(5-16)-Cys-X(2)-Cys-X(6-8)-Cys characteristic of the Zn(II)(2)Cys(6) transcriptional factors. YNA2 is therefore a member of the H. polymorpha nitrate assimilation gene cluster which is transcribed in the opposite direction to the rest of the members. Yna2p shares about 27% similarity with the H. polymorpha Yna1p Zn(II)(2)Cys(6) transcriptional factor involved in nitrate induction. Unlike the wild-type, the yna2::URA3 strain showed no expression of the nitrate assimilation genes when incubated in nitrate for 2 h. With regard to YNA2 expression, similar YNA2 transcript levels were observed in ammonium and in ammonium plus nitrate, but about a four-fold higher expression was observed in nitrate. However, this induction by nitrate of the YNA2 gene was not observed in the Deltayna1::URA3 strain. On the contrary, the pattern of YNA1 expression was the same in the wild-type as in the yna2::URA3 strain, indicating that YNA2 does not affect YNA1 expression. PMID- 11921103 TI - Occurrence of 20S RNA and 23S RNA replicons in industrial yeast strains and their variation under nutritional stress conditions. AB - We have characterized industrial yeast strains used in the brewing, baking, and winemaking industries for the presence or absence of cytoplasmic single-stranded 20S and 23S RNAs. Furthermore, the variation of intracellular concentrations of these replicons in brewing and laboratory strains under nutritional stress conditions was determined. Our results show a correlation between the relative abundance of these replicons and exposure of yeast to nutritionally stressful conditions, indicating that these RNAs could be employed as molecular probes to evaluate the exposure of 20S(+) and/or 23S(+) yeast strains to stress situations during industrial manipulation. During this study, several 20S(-)23S(+) Saccharomyces cerevisiae strains were isolated and identified. This is the first time that a yeast strain containing only 23S RNA has been reported, demonstrating that 20S RNA is not required for 23S RNA replication. PMID- 11921104 TI - Chromosome V loss due to centromere knockout or MAD2-deletion is immediately followed by restitution of homozygous diploidy in Saccharomyces cerevisiae. AB - To investigate the possibility of inducing specific chromosome loss by centromere deletion in eukaryotic cells, the yeast diploid strain ZG1, carrying three pairs of heterozygous marker genes (CAN1(S)/can1(R), URA3/Deltaura3, hphMX4/HIS1), widely spread on both arms of chromosome V, was constructed. One of the two centromeres V of ZG1 was replaced by the LEU2 gene via the well-established PCR mediated knockout technique. After DNA transformation, putative yeast colonies that showed loss of heterozygosity (LOH) for the three markers of chromosome V (CAN1(S) URA3 hphMX4) were identified among the colonies selected for leucine prototrophy. Phenotypic tests, colony-PCR and Southern blot analysis of these cells demonstrated the physical loss of the CAN1(s), URA3, and hphMX4 marker genes from the genome. Further tetrad analysis results were consistent with this conclusion; however, four-spore viability indicated a normal chromosome number of these transformants. To verify the diploidy of the selected chromosome V, the HIS1 gene was deleted with a standard KanMX4 knockout DNA cassette. The resulting heterogeneity of the HIS1/KanMX4 markers, together with quantitative PCR and densitometric analysis on chromosome V, confirmed its diploid complement, thereby indicating that an endoreduplication event had taken place. Restitution of diploidy also occurred in MAD2-deleted strains undergoing higher rates of spontaneous chromosome V loss, indicating a more general phenomenon that is undetectable by phenotypic analysis alone. PMID- 11921105 TI - Current awareness on yeast. PMID- 11921106 TI - Is it time to abandon functional imaging in the study of neuroprotection? PMID- 11921107 TI - Repairing the brain in Parkinson's disease: where next? PMID- 11921108 TI - Prevalence of Parkinson's disease in Cantalejo, Spain: a door-to-door survey. AB - We assessed the prevalence of Parkinson's disease in Cantalejo, Spain. In 1994, we screened 1,579 persons (age > or = 40 years) using a high-sensitivity method. Cases fulfilling established clinical criteria were followed for a minimum of 3 years. Prevalences were compared with those from other door-to-door surveys. We detected 27 individuals with parkinsonism, 20 of whom had Parkinson's disease. The prevalence of Parkinson's disease increased with age and, when age-adjusted to European standards, was 9.01 per 1,000 (age 40 years and over; 10.78 in men and 5.23 in women). Of the 11 men, three were in Hoehn & Yahr grades III-IV, but six of the nine women were more severely affected. Overall, we found 18 newly diagnosed cases of parkinsonism, 13 of which were Parkinson's disease, and the majority of which were in men aged 80 years or older with a mean duration of illness of 5 years. Our prevalence figures are the highest reported, apparently because of the inclusion of several very elderly men. Parkinson's disease in Cantalejo is less severe in men than in women, particularly in those newly diagnosed. Despite the low numbers, the high prevalence and sex-related pattern are unexplained but they probably relate to the high sensitivity of the screening method. PMID- 11921109 TI - Do risk factors for Alzheimer's disease predict dementia in Parkinson's disease? An exploratory study. AB - The extent to which concomitant Alzheimer's disease (AD) is etiologically related to the development of dementia in Parkinson's disease (PD) remains controversial. We explored the association of four risk factors associated with AD, including head injury, smoking, hypertension, and diabetes mellitus, with incident dementia in PD. A cohort of 180 nondemented PD patients from the Washington Heights community in northern Manhattan, New York, completed a risk factor questionnaire at baseline and was followed annually with neurological and neuropsychological evaluations. The association of baseline variables with incident dementia was analyzed by using Cox proportional hazards models. All analyses controlled for age at baseline, gender, years of education, duration of PD, and total Unified Parkinson's Disease Rating Scale (UPDRS) motor score at baseline. Of 180 patients (mean age, 71.0 +/- 10.3 years), 52 (29%) became demented during a mean follow-up period of 3.6 +/- 2.2 years. Head injury risk ratio ([RR] 0.9; 95% confidence interval [CI], 0.4-2.2; P = 0.9), hypertension (RR, 0.7; 95% CI, 0.4-1.4, P = 0.3), and diabetes mellitus (RR, 0.8; 95% CI, 0.3-2.3; P = 0.7) were not significantly associated with incident dementia in the Cox models. Patients who reported having ever smoked were at increased risk for the development of dementia compared with nonsmokers (RR, 2.0; 95% CI, 1.0-3.9; P = 0.05). Current smoking was significantly associated with incident dementia (RR, 4.5; 95% CI, 1.2 16.4; P = 0.02), whereas past smoking approached significance (RR, 1.9; 95% CI, 0.9-3.7; P = 0.07). Although an inverse association between smoking and PD has been reported in several studies, our study showed a positive association between smoking and dementia in the setting of PD. The association of smoking with incident dementia in PD deserves further study. PMID- 11921110 TI - Field validation of a method for population screening of parkinsonism. AB - We sought to validate an instrument and two application procedures for screening for prevalence of parkinsonism in conjunction with a door-to-door survey in Cantalejo, Spain. Residents in Cantalejo aged 40 years or older were assigned to three groups specifically designed to optimise case-finding and cost/efficiency. A nine-item questionnaire aimed at identifying parkinsonism-related symptoms was administered and collected door-to-door by laymen and repeated by specialists at a medical facility before neurological examination. Diagnoses were then established and confirmed after a 3-year follow-up. Different indices for and concordance of answers were studied for both applications in groups of 25 individuals with parkinsonism. The scoring procedure was updated using the prevalence community sample of parkinsonism. Whereas the application administered by laymen showed, using the same cut-off scores, a higher sensitivity for parkinsonism than that administered by trained personnel, it yielded a lower positive predictive value for parkinsonism diagnosed during the study, i.e., at a 30-point cut-off (96% vs. 80% and 15% vs. 76%, respectively). Concordance was > or =64% with kappa >or=0.316 and P 0.10). As controversy continues regarding the medicated treatment of PLMS, our findings with selegiline point to an alternative for clinicians to consider. PMID- 11921130 TI - Novel Italian family supports clinical and genetic heterogeneity of primary adult onset torsion dystonia. AB - We report on an Italian kindred with adult-onset primary torsion dystonia (PTD). A detailed clinical examination of the six definitely affected family members revealed a mild, purely focal phenotype. The disease involved only one body part (eyes, neck, or arm). PTD in this family was not linked to the known disease loci (DYT1, DYT6, DYT7, and DYT13), and the 3-bp deletion in the DYT1 gene was also excluded. These findings support genetic heterogeneity of PTD and indicate that a novel unassigned gene is responsible for focal dystonia in this family. PMID- 11921132 TI - Dopa-responsive parkinsonism secondary to right temporal lobe haemorrahage. AB - A 46-year-old man developed a symmetrical parkinsonian syndrome 7 weeks after large right temporal intracerebral haemorrhage resulting from a ruptured arteriovenous malformation. His signs included bradykinesia, rigidity, start hesitation, and poor postural reflexes, without a resting tremor. He also had signs of a Parinaud's syndrome. Computed tomography and magnetic resonance imaging of the brain demonstrated changes in the right temporal lobe associated with the haemorrhage but no abnormality of the basal ganglia or midbrain. Levodopa therapy produced a dramatic improvement within a few days of commencement. We postulate that the parkinsonism resulted from midbrain compression secondary to transtentorial herniation. Although parkinsonism is a rare complication of lobar intracerebral haemorrhage, it is important to recognise as it may be potentially treatable. PMID- 11921133 TI - Thalamic stimulation for midbrain tremor after partial hemangioma resection. AB - We describe a patient with disabling medication-resistant midbrain tremor developed after partial hemangioma resection, who responded to deep brain stimulation of the thalamus. PMID- 11921134 TI - Frequency of DYT1 mutation in early onset primary dystonia in Italian patients. AB - Thirty Italian patients with sporadic, early-onset, primary dystonia were screened for the DYT1 mutation. Five patients were positive (mean age at onset, 8 years); two had the typical phenotype, two a generalised dystonia also involving the cranial muscles, and one a segmental dystonia. In the other 25 patients (mean age at onset, 7.7 years), dystonia was generalised in 22 patients and remained segmental in three. Our results indicate the role of DYT1 mutation in Italian patients and confirm clinical and genetic heterogeneity of early-onset primary dystonia. PMID- 11921135 TI - Myoclonic status epilepticus: video presentation. AB - A young woman with juvenile myoclonic epilepsy had recurrent attacks of myoclonic status epilepticus related to a long history of limited compliance and irregular sleep. The diagnosis of this clinical pattern is based mainly on clinical description. A home video captured an attack. PMID- 11921136 TI - Suppression of cortical myoclonus by levetiracetam. AB - A 16-year-old boy suffered severely disabling posthypoxic myoclonus. Neurophysiological investigation showed cortical but not reticular reflex myoclonus. Add-on therapy with levetiracetam significantly improved the patient's clinical condition, suppressed cortical myoclonus-associated spikes, and enabled further neurorehabilitation. PMID- 11921137 TI - Orofacial dyskinesias in a patient with primary biliary cirrhosis: a clinicopathological case report and review. AB - We describe the pathological and clinical aspects, including video and radiological magnetic resonance imaging, of a case of chronic acquired hepatocerebral degeneration with orofacial dyskinesias in relation to a primary biliary cirrhosis. We provide a review of the literature on this subject. PMID- 11921138 TI - Association of chorea and motor neuron disease. AB - Amyotrophic lateral sclerosis (ALS) is classically characterized by the presence of symptoms or signs of upper and lower motor neuron impairment and sparing of other neuronal systems.1 We report on a patient who was primarily diagnosed as typical ALS and developed chorea 10 years after the onset of motor neuron signs. PMID- 11921139 TI - Entacapone in restless legs syndrome. AB - Entacapone increased the duration of action of carbidopa-levodopa and resulted in longer periods of symptomatic relief in a patient with restless legs syndrome. The only side effect was nausea. PMID- 11921140 TI - Woman with facial spasm induced exclusively by sound. AB - A woman is described in whom facial spasm occurs solely and predictably in the presence of certain noises. The symptoms resolve promptly as soon as the auditory stimulus stops. PMID- 11921141 TI - Phenotypic variability in a large kindred (Family LA) with deletions in the parkin gene. PMID- 11921144 TI - Deep brain stimulation in Huntington's disease. PMID- 11921143 TI - Antiparkinsonian treatment in pregnancy. PMID- 11921146 TI - Isolated hallucinosis in Alzheimer's disease is associated with African-American race. AB - OBJECTIVES: The aim of this investigation was to study the relationship between isolated hallucinosis and race in Alzheimer's disease. METHODS: This was a cross sectional, case control study carried out at the Neuropsychiatry Service, outpatient clinic at the Johns Hopkins School of Medicine, USA. The participants were 237 community-residing patients with probable Alzheimer's disease according to NINCDS/ADRDA criteria were included in the study. 9 patients with isolated hallucinosis were compared to a control group of 228 patients who had neither delusions nor hallucinations. Patients with only delusions or both delusions and hallucinations were excluded based on prior research. Patients were assessed clinically for the presence of hallucinations using the DSM-IV glossary definitions. They were also rated on standardized measures of cognitive impairment, depression, functional impairment, and general health. RESULTS: There was a significant association between hallucinations and race in patients with Alzheimer's disease. Before adjustment for other variables, the African-American race conferred a 5.5-fold (95% CI = 1.4-21.6; p = 0.02) increased risk for isolated hallucinosis. After adjustment for multiple other variables, this risk increased further to 27.2-fold (95% CI = 1.6-457.3; p = 0.02). CONCLUSIONS: African-American patients with Alzheimer's disease are more likely to have isolated hallucinations than Caucasian patients even after statistical adjustment for multiple confounding variables, which might distort this association. This finding has implications for our understanding of the etio-pathogenesis of hallucinations in Alzheimer's disease and for meeting health service needs of African-American patients. PMID- 11921147 TI - Reducing the burden of caring for Alzheimer's disease through the amelioration of "delusions of theft" by drug therapy. AB - BACKGROUND: Delusions of theft (delusions involving the theft of possessions) are one of the most frequent neuropsychiatric manifestations of Alzheimer's disease (AD). OBJECTIVE: The current study investigated the presence and extent of such delusions before and after drug treatment in a group of AD patients, and the consequent effects on the burden of care on caregivers. METHOD: The study was an open-label cohort design. The delusions studied consisted only of those involving theft of possessions. Sixteen AD patients served as subjects in order to assess the efficacy of Risperidone administration, in the reduction or elimination of these delusions. The caregiver burden was evaluated using the Zarit Caregiver Burden Interview (ZBI) before the administration of Risperidone and 12 weeks after administration, for cases where delusions of theft were eliminated or reduced. RESULTS: The burden of care on caregivers was significantly reduced (p < 0.001) through the elimination or reduction of delusions of theft. CONCLUSION: Delusions of theft are considered to be a major factor in increasing the burden of care, and the treatment of these, through appropriate drug therapy, is therefore of great importance in the continuation of satisfactory care in the home. PMID- 11921148 TI - What are patients and their families told about the diagnosis of dementia? Results of a family survey. AB - BACKGROUND: Controversy exists as to whether dementia patients should be told their diagnosis and prognosis. OBJECTIVE: This study examined the experience of patients and families when a diagnosis of dementia is given. METHOD: Fifty-seven family members from community dementia support groups answered a questionnaire regarding the diagnosis of dementia in a family member. RESULTS: Family members were significantly more likely to have been told the diagnosis and symptoms to expect in dementia than patients themselves. Half of the families felt they were not given enough information regarding dementia. Interestingly, the majority of family members believed patients should be told their diagnosis and prognosis yet about half had reported that informed patients had reacted poorly to being told their diagnosis and only about a third felt it was helpful to the patient. CONCLUSION: The results suggest physicians should better involve patients and their families in disclosing diagnoses and information regarding dementia. The results also suggest the current AMA guidelines to inform patients their diagnosis of dementia are inadequate to address the clinical complexities of this issue. PMID- 11921149 TI - Revealing a hidden problem. An evaluation of a community dementia case-finding program from the Indian 10/66 dementia research network. AB - BACKGROUND: Dementia in India is largely a hidden problem with no community awareness and little help seeking from affected families, despite high levels of strain. Cases must therefore be identified before practical help can be offered. METHOD: After two and a half hours of formal training, local community health workers in rural Kerala were asked to identify possible cases of dementia from the community they served. Diagnoses were then verified by a senior local psychiatrist with clinical and research interests in old age psychiatry. RESULTS: The community health workers identified 51 out of 1979 over 60 year old residents (a prevalence of 2.6%) as suspected cases of dementia. Following the psychiatrist's assessment, 33 met DSM-IV criteria for dementia. The majority of confirmed cases were of the Alzheimer's Disease sub-type. Most "non-cases" were found to be suffering from other major psychiatric disorders, with substantial unmet need. The positive predictive value of the community health workers informal screening was 64.7%. CONCLUSIONS: This simple cost-effective case finding method can be of practical use in the development of community based dementia care services in India and other developing countries with similar health care systems. PMID- 11921150 TI - Does the HoNOS 65+ meet the criteria for a clinical outcome indicator for mental health services for older people? AB - BACKGROUND: A clinical indicator should demonstrate clinically meaningful change, be relevant, allow comparisons between services, be acceptable to clinicians, and have acceptable validity, reliability and sensitivity to change. The HoNOS 65+ has been suggested as a clinical outcome indicator. The sensitivity to change of the HoNOS 65+ is not known. METHODS: This is a prospective study using routine clinical data. A pilot cohort (n = 42) was used to measure the concurrent validity of the HoNOS 65+ with the Mini-Mental State Examination (MMSE), Geriatric Depression Scale (GDS-15) and Brief Agitation Rating Scale (BARS). The main cohort of 245 consecutive referrals to a community mental health service for older adults was used to assess sensitivity to change against the CIBIC+. RESULTS: The HoNOS 65+ was acceptable to case managers, most HoNOS 65+ items had excellent interrater reliability and the HoNOS 65+ had good concurrent validity. Changes in the HoNOS 65+ scores between assessment and discharge had a moderate, but significant correlation with CIBIC+ scores. CONCLUSION: The HoNOS 65+ meets the criteria for a clinical outcome indicator for community mental health services for older people. The HoNOS 65+ is sensitive to change. PMID- 11921151 TI - Hyponatraemia in elderly psychiatric patients treated with Selective Serotonin Reuptake Inhibitors and venlafaxine: a retrospective controlled study in an inpatient unit. AB - OBJECTIVE: To determine the prevalence of hyponatraemia associated with selective serotonin reuptake inhibitor (SSRI) and venlafaxine use in elderly patients compared to that in elderly patients not prescribed these drugs, while controlling for age, sex, depression status and illnesses or prescribed medications also associated with hyponatraemia. Design and setting Retrospective controlled analysis in a 36-bed inpatient unit for elderly psychiatric patients in Melbourne. PATIENTS: Inpatients (199) with a mean age of 74.2 years of whom 74 were prescribed an SSRI or venlafaxine. RESULTS: Patients on SSRIs or venlafaxine were 5.6 times as likely as patients not so treated to have hyponatraemia. Thirty nine percent of patients on an SSRI or venlafaxine had hyponatraemia compared with 10% of controls. Ten of the 14 patients on venlafaxine were hyponatraemic. Controlling for thiazide status did not reduce the odds of these patients having hyponatraemia and taking an SSRI or venlafaxine was still strongly associated with hyponatraemia after also controlling for age, sex, and depression status, consumption of other drugs potentially causative of hyponatraemia and medical illness severity (Odds Ratio (OR) 3.5, p = 0.008). CONCLUSIONS: SSRI and venlafaxine use is strongly associated with the presence of hyponatraemia in a population of elderly psychiatric inpatients and the association is not due to confounding by age, sex, depression status, medical illness severity or consumption of other drugs. Elderly patients on SSRIs or venlfaxine should have sodium levels checked before and after commencement of antidepressant treatment. PMID- 11921152 TI - Psychosocial and clinical predictors of unipolar depression outcome in older adults. AB - BACKGROUND: This study examined psychosocial and clinical predictors of depression non-remittance among a sample of initially clinically depressed elders. METHODS: Incident and prevalent unipolar depression cases (n = 166) were enrolled into the MHCRC for the Study of Depression in Late Life and followed for 12 months while undergoing treatment using a standardized algorithm. The outcome was remission vs non-remission (<6 vs > 7 on the Montgomery-Asberg Depression Rating Scale (MADRS)) at one-year follow-up. Baseline predictor variables included psychosocial factors, such as four domains of social support, basic and instrumental activities of daily living (ADLs), and clinical factors, which included use of ECT, past history of depression, comorbidities, and antidepressant treatment. RESULTS: At one-year follow-up, 45% of the sample was in remission based upon MADRS scores. In bivariate analyses, non-remitted patients were more likely at baseline to use benzodiazepines, anxiolytic/sedatives, and/or MAO inhibitors than patients in remission, and have more depressive episodes. Among psychosocial factors, non-remitted patients had at baseline, more ADL and IADL problems and decreased subjective social support as compared to patients in remission. In logistic regression analyses more depression episodes, using anxiolytic/sedatives, more IADL problems and decreased subjective social support predicted poor depression outcome after one-year. CONCLUSIONS: While clinical and diagnostic variables were related to improvement, baseline psychosocial factors were also important. PMID- 11921154 TI - Psychometric properties of Clock Drawing Test and MMSE or Short Performance Test (SKT) in dementia screening in a memory clinic population. AB - OBJECTIVE: To evaluate five different scoring methods of the Clock Drawing Test (CDT) and to examine whether a combination of Mini Mental State Examination (MMSE) or Short Performance Test (Syndrom Kurz Test, SKT), respectively, with CDT can be used for cognitive screening. METHODS: Retrospective blinded analysis of clock drawing performance using five scoring methods (Shulman et al. (1986), Sunderland et al. (1989), Wolf-Klein et al. (1989), Watson et al. (1997), Manos (1997)). A Memory Clinic at an academic psychiatric hospital (University of Frankfurt am Main, Germany). 123 consecutive patients (79 dementia patients, 44 controls). Inter-rater reliability and correlation of five different scoring methods of the CDT with established psychometric tests. Sensitivity and specificity of all five CDT's using the original and modified cut-off scores. Sensitivity, specificity and positive and negative predictive value of a combination of the CDT with MMSE and SKT, respectively. RESULTS: All scoring methods of the CDT showed a highly significant interrater reliability (0.82 to 0.94). Correlation with the MMSE and the SKT was also significant (p < 0.01) for all five CDTs. Highest sensitivity was achieved by the Shulman scoring method (81% sensitivity, specificity 79%). Sensitivity of all scoring methods could be improved up to 89% by modifying the originally proposed cut-off scores at the cost of lower specificity. By combining the CDT with the MMSE or the SKT, respectively, the sensitivity of each of the tests could be improved to 92% (SKT and Shulman scale). In patients with mild dementia (GDS 3), a combination of the Shulman Scale with the SKT (92%) and the MMSE (75%) achieved the highest sensitivity. CONCLUSIONS: The CDT in combination with the MMSE or SKT is an easily administered, non threatening and highly sensitive screening test for dementia in the setting of a memory clinic. PMID- 11921153 TI - Incidence, prevalence, and outcomes of depression in residents of a long-term care facility with dementia. AB - BACKGROUND: The course of depression in residents of long-term care with dementia is not well studied. OBJECTIVES: To estimate the incidence, prevalence, and outcomes of depression in long-term care residents with dementia. METHOD: 201 residents of Copper Ridge, a long-term care facility for the memory impaired, were followed every six months during the first year after their admission. RESULTS: On admission 19.9% of the residents had depression, the majority (75%) occurring in persons with a prior history of depression. At six months, only 15% of the original 40 depressed patients were still depressed, while at twelve months only 7.5% were depressed. The incidence of depression at six and twelve months was 1.8% and 6.4% respectively. Most persons with new depression at six months were no longer depressed at one year. The annual attack rate (cumulative likelihood of depression over one year) for the total population was 26.4%. The rates for the subgroup of Alzheimer's were similar to the rates of the total population, except for an annual attack rate of 17.5%, reflecting a lower rate of depression on admission. The decline in depression over the year after admission is noteworthy and likely reflects appropriate diagnosis and treatment of depression. CONCLUSIONS: These results indicate that, over the course of one year, depression afflicts a considerable proportion of long-term care residents with dementia. Admission to a long-term care facility may actually result in a reduction of depression within the year after admission in part due to rapid recognition, appropriate diagnosis and treatment. PMID- 11921155 TI - A study of suicides of older people in Sydney. AB - BACKGROUND: Reports based on studies of coroner's files show that suicides in old age are commonly related to depression, but that in a majority of cases disability or ill-health is also a major factor. The aim of this study was to try to understand more clearly the precipitant causes of suicide in an older population. METHOD: An Australian metropolitan coroner's office provided data concerning suicides in 1994-1998 of persons aged over 65 years. RESULTS: Of 210 elderly people who killed themselves, 160 (76%) were clearly depressed, including a majority of the 24% deemed to have understandable reasons for suicide. Physical ill-health or disability was the major factor leading to suicide in 34% and appeared to contribute to suicidal ideation in another 24% of those who died; they had usually not been seen by psychiatrists. CONCLUSION: Because depression is often treatable, even when associated with depressing circumstances, there is potential for further reduction of old age suicide rates by recognising and appropriately responding to symptoms of depression and distress. PMID- 11921156 TI - A 24-week, double-blind, placebo-controlled trial of donepezil in patients with Down syndrome and Alzheimer's disease--pilot study. AB - BACKGROUND: Donepezil Hydrochloride (Aricept) is a selective anticholinesterase inhibitor developed for the treatment of Alzheimer's disease (AD). This study investigated the safety and efficacy of the drug to treat Down syndrome (DS) adults with mild to moderate AD. METHOD: This was a 24-week, double blind, placebo controlled, parallel-group trial. Patients were randomized to receive placebo or donepezil (5 mg per day during the first four weeks, and then 10 mg per day thereafter). Primary efficacy was measured using the Dementia Scale for Mentally Retarded Persons (DMR), and secondary efficacy was measured using the Severe Impairment Battery (SIB), Neuropsychiatric Inventory (NPI) and by the Adaptive Behavior Scale (ABS). RESULTS: A total of 30 DS patients with AD entered the study of which 27 were included in the subsequent data analysis. The donepezil group had non-statistically significant reduction in deterioration in DMR, SIB, and ABS mean scores relative to the placebo group. However NPI scores showed less improvement in the donepezil group when compared to the placebo group. Fifty percent of subjects in the donepezil group showed improvement in mean DMR scores at the end point compared to baseline, when compared to 31% on placebo. There were no life threatening adverse effects associated with treating adults with DS with donepezil. A number of side-effects did occur including diarrhoea, insomnia, fatigue, and nausea. CONCLUSION: Donepezil Hydrochloride administered once a day appears to be generally well tolerated and safe in DS adults who have AD. There is some possible efficacy in the treatment of symptoms of mild to moderate Alzheimer's disease in this population, although the sample size of this study was too small for statistical significance. It is recommended that donepezil, with the appropriate precautions, should be considered for the treatment of AD in adults with DS as deemed by a specialist. PMID- 11921158 TI - Switching cholinesterase inhibitor therapy in Alzheimer's disease--donepezil to rivastigmine, is it worth it? PMID- 11921157 TI - Validation of the Spanish version of the geriatric depression scale (GDS) in primary care. AB - OBJECTIVE: To estimate the predictive value of the 30-question Geriatric Depression Scale (GDS) in Spanish and calculate the most adequate cut-point for its use in Primary Health Care consultations. METHOD: 218 patients over the age of 64 treated at three health centers of Area 10 in Madrid were selected. In the first phase, the subjects completed the GDS, the Mini-Mental State Examination (MMSE) and a questionnaire on health and socio-demographic variables. They were later interviewed using the Geriatric Mental Schedule (GMS), used as the gold standard by doctors who were unaware of the results of the GDS. Two categories were contemplated according to the results of the GMS: cases of depression (diagnosis of psychotic or neurotic depression) and non-psychiatric cases (no psychiatric diagnosis, although isolated symptoms could be present). RESULTS: 192 aged subjects were interviewed using the GDS and the GMS. Of these, 103 were considered "non-cases of depression" and 60 others made up the "cases of psychotic/neurotic depression" group. For the most effective cut-point (9/10), sensitivity was 86.7% and specificity 63.1%. Considering a prevalence of depression of 30%, the predictive value for positives was 50.2% and for negatives 91.7%. The Cronbach alpha coefficient was 0.82, and the area below the ROC curve obtained was 0.85. Those patients with cognitive deterioration had a mean GDS score similar to those that did not present deterioration (11.16 vs 10.52; p > 0.05). CONCLUSIONS: The Geriatric Depression Scale is valid as a screening test in Primary Care consultations due to its high sensitivity and negative predictive value. The most effective Spanish GDS cut-point (9/10) is lower than that obtained in the original English version (10/11). PMID- 11921159 TI - Management of patients with cognitive impairment by residential doctors. PMID- 11921162 TI - Current awareness. PMID- 11921163 TI - Take five: a myosin class act in fission yeast. PMID- 11921164 TI - Expression of constructs of the neuronal isoform of myosin-Va interferes with the distribution of melanosomes and other vesicles in melanoma cells. AB - Myosin-Va has been implicated in melanosome translocation, but the exact molecular mechanisms underlying this function are not known. In the dilute, S91 melanoma cells, melanosomes move to the cell periphery but do not accumulate in the tips of dendrites as occurs in wild-type B16 melanocytes; rather, they return and accumulate primarily at the pericentrosomal region in a microtubule-dependent manner. Expression of the full-length neuronal isoform of myosin-Va in S91 cells causes melanosomes to disperse, occupying a cellular area approximately twice that observed in non-transfected cells, suggesting a partial rescue of the dilute phenotype. Overexpression of the full tail domain in S91 cells is not sufficient to induce melanosome dispersion, rather it causes melanosomal clumping. Overexpression of the head and head-neck domains of myosin-Va in B16 cells does not alter the melanosome distribution. However, overexpression of the full tail domain in these cells induces melanosome aggregation and the appearance of tail associated, aggregated particles or vesicular structures that exhibit variable degrees of staining for melanosomal and Golgi beta-COP markers, as well as colocalization with the endogenous myosin-Va. Altogether, the present data suggest that myosin-Va plays a role in regulating the direction of microtubule dependent melanosome translocation, in addition to promoting the capture of melanosomes at the cell periphery as suggested by previous studies. These studies also reinforce the notion that myosin-V has a broader function in melanocytes by acting on vesicular targeting or intracellular protein trafficking. PMID- 11921165 TI - Regulation of focal adhesion targeting and inhibitory functions of the FAK related protein FRNK using a novel estrogen receptor "switch". AB - Focal adhesion kinase (FAK) is a regulator of numerous adhesion-dependent processes including cell migration, cell proliferation, and cell survival. The C terminal domain of FAK, FAK-related nonkinase (FRNK), is autonomously expressed and functions as an inhibitor of FAK signaling. Previous attempts to use FRNK as a tool to dissect FAK signaling have been limited because of an inability to temporally regulate the inhibitory functions of FRNK. In this report, we describe and characterize a conditionally targeted form of FRNK that was created by fusing the hormone-binding domain of the estrogen receptor (ER*) to the C-terminus of FRNK. In the absence of added hormone, FRNK-ER* was diffusely distributed throughout the cytoplasm of cells. Upon addition of hormone, the cytoplasmic pool of FRNK-ER* was rapidly redistributed to focal adhesions. We demonstrate that cells expressing FRNK-ER* show a hormone-dependent decrease in FAK tyrosine phosphorylation and cell migration. Furthermore, when cells expressing of FRNK ER* were treated with hormone, the cells responded with a dramatic change in cell morphology, suggesting a role for FAK in the regulation of the adhesive properties of focal adhesions. PMID- 11921166 TI - Axonemal activity relative to the 2D/3D-waveform conversion of the flagellum. AB - The waveform of the flagellum of the sea urchin spermatozoon is mainly planar, but its 3D-properties were evoked for dynamic reasons and described as helical. In 1975, the apparent twisting pattern of the sea urchin axoneme was described [Gibbons I. 1975. The molecular basis of flagellar motility in sea urchin spermatozoa. In: Inoue S, Stephens R, editors. Molecular and cellular movement. New York: Raven Press, p. 207-232.] and was considered to be one of the main elements involved in axonemal behaviour. Recently, planar, quasi-planar, and helical waveforms were observed when the flagellum of sea urchin sperm cells was submitted to an increase in viscosity. The quasi-planar conformation seemed to be due to the alternating torsion of the inter-bend segments [Woolley D, Vernon G. 2001. A study of helical and planar waves on sea urchin sperm flagella, with a theory of how they are generated. J. Exp. Biol. 204:1333-1345]. These three waveforms, which are due to a change in axonemal activity, are possibly used by the sperm cells to adapt their movement to variations in the physico-chemical characteristics of the medium (seawater) in which the cells normally swim. We constructed a simple model to describe qualitatively the central shear (between the axonemal doublets and the central pair) and the tangential shear (between the doublets themselves). In this model, the 3D-bending is resolved into components in two perpendicular planes and each of the nine planes of inter-doublet interaction defines a potential bending plane that is independently regulated. These shears were calculated for the three waveforms and their inter-conversion. This allowed us to propose that axoneme is resolved in successive modules delineated by abscissas where the sliding is always nil. We discuss these data concerning the axonemal machinery, and especially the alternating activity of opposite sides of (two) neutral surface(s) that seem(s) to be responsible for this inter-conversion, and for the possible twist of the axoneme during the beating. PMID- 11921168 TI - Spatial and temporal regulation of actin polymerization for cytoskeleton formation through Arp2/3 complex and WASP/WAVE proteins. PMID- 11921169 TI - Post-karyokinesis centrosome movement leaves a trail of unanswered questions. AB - The centrosome is a complex structure composed of a large number of proteins (pericentriolar material, PCM) usually organized around a pair of centrioles (or a centriole duplex). This structure is capable of nucleating and organizing microtubules, duplication, and motility. In general, episodes of dramatic centrosome movement correlate with periods of cellular reorganization and nowhere is cellular reorganization more apparent, or more important, than in the periods before and after cell division. It is now clear that centrosome movement occurs not only prior to cell division but also at its completion, in concert with cytokinesis. The focus of this review is the newly emerging picture of centrosome activity during the post-karyokinesis period and the role that this activity might play in the transition of cells from mitosis to interphase. PMID- 11921170 TI - Regulators of GTP-binding proteins cause morphological changes in the vacuole system of the filamentous fungus, Pisolithus tinctorius. AB - Tubule formation is a widespread feature of the endomembrane system of eukaryotic cells, serving as an alternative to the better-known transport process of vesicular shuttling. In filamentous fungi, tubule formation by vacuoles is particularly pronounced, but little is known of its regulation. Using the hyphae of the basidiomycete Pisolithus tinctorius as our test system, we have investigated the effects of four drugs whose modulation, in animal cells, of the tubule/vesicle equilibrium is believed to be due to the altered activity of a GTP binding protein (GTP gamma S, GDP beta S, aluminium fluoride, and Brefeldin A). In Pisolithus tinctorius, GTP gamma S, a non-hydrolysable form of GTP, strongly promoted vacuolar tubule formation in the tip cell and next four cells. The effects of GTP gamma S could be antagonised by pre-treatment of hyphae with GDP beta S, a non-phosphorylatable form of GDP. These results support the idea that a GTP-binding protein plays a regulatory role in vacuolar tubule formation. This could be a dynamin-like GTP-ase, since GTP gamma S-stimulated tubule formation has only been reported previously in cases where a dynamin is involved. Treatment with aluminium fluoride stimulated vacuolar tubule formation at a distance from the tip cell, but NaF controls indicated that this was not a GTP-binding-protein specific effect. Brefeldin A antagonised GTP gamma S, and inhibited tubule formation in the tip cell. Given that Brefeldin A also affects the ER and Golgi bodies of Pisolithus tinctorius, as shown previously, it is not clear yet whether the effects of Brefeldin A on the vacuole system are direct or indirect. PMID- 11921173 TI - Recent trends in breast cancer incidence and mortality. AB - Breast cancer accounts for one-third of cancer diagnoses and 15% of cancer deaths in U.S. women. Its 192,000 cases and 40,000 deaths in 2001 make it the most common incident cancer (excluding superficial skin cancers) and second leading cause of cancer death. Over one-half of the 300,000 breast cancer deaths worldwide in 1990 (the latest year with such data) occurred in developed countries, but annual mortality rates ranged from 27/100,000 women in northern Europe to 4/100,000 women in Asia. Incidence data are less complete, although 1988-1992 rates varied threefold: low in Asia, intermediate in South America and Eastern Europe, and high in North America and Western Europe. Migrant studies suggest that lifestyle factors largely explain these international differences. U.S. incidence rates are generally 20%-40% higher in white women than in non white women, but are higher in young (under age 40) black women than in young white women. Incidence rates rose in the 1970s, leveled off in the 1990s, and are declining for young women. Women in some areas of the northeast U.S. have twofold higher mortality than that of other U.S. women, but reproductive and socioeconomic characteristics explain much of that difference. In the 1970s and 1980s, mortality rates held steady in developed countries but rose in developing countries. Since 1987 mortality rates fell by 25% as a result of earlier detection and improved treatment. Age-period-cohort analyses indicate that changes in recognized risk factors may affect mortality patterns. Continued analysis of international and intranational trends may reveal targets for multidisciplinary intervention and prevention efforts. PMID- 11921171 TI - A human homologue of Drosophila kelch associates with myosin-VIIa in specialized adhesion junctions. AB - Mutations in myosin-VIIa are responsible for the deaf-blindness, Usher disease. Myosin-VIIa is also highly expressed in testis, where it is associated with specialized adhesion plaques termed ectoplasmic specializations (ES) that form between Sertoli cells and germ cells. To identify new roles for myosin-VIIa, we undertook a yeast two-hybrid screen to identify proteins associated with myosin VIIa in the ES. We identified Keap1, a human homologue of the Drosophila ring canal protein, kelch. The kelch-repeats in the C-terminus of human Keap1 associate with the SH3 domain of myosin-VIIa. Immunolocalization studies revealed that Keap1 is present with myosin-VIIa in the actin bundles of the ES. Myosin VIIa and Keap1 copurify with ES and colocate with each other and with F-actin at the electron microscopy level. Interestingly, in many epithelial cell types including cells derived from retina and inner ear, Keap1 is a component of focal adhesions and zipper junctions. Keap1 can target to the ES in the absence of myosin-VIIa, suggesting that Keap1 associates with other molecules in the adhesion plaque. Keap1 and myosin-VIIa overlapped in expression in the inner hair cells of the cochlea, suggesting that Keap1 may be a part of a family of actin binding proteins that could be important for myosin-VIIa function in testis and inner ear. PMID- 11921174 TI - Smoking (active and passive) and breast cancer: epidemiologic evidence up to June 2001. AB - The first generation of studies evaluating the association between exposure to tobacco smoke and breast cancer merely compared active to nonactive smokers, with varying degrees of detail in the definition of active smoking. With rare exceptions, studies of this kind failed to show an effect of smoking on breast cancer risk. However, such analysis is probably insufficient. The most recent reports on the smoking-breast cancer connection have two characteristics. Some have separated women exposed to passive smoking from those nonexposed to either active or passive smoke. Other reports have focused on factors that modify the effect of smoking on breast cancer incidence, such as genetic markers or hormone receptors. A minority of reports combines these two characteristics. This review addresses the epidemiologic evidence for a link between smoking and breast cancer and discusses the implications of this evidence for future studies. PMID- 11921175 TI - Making epidemiologic studies responsive to the needs of participants and communities: the Carolina Breast Cancer Study experience. AB - In this report, we present the results of surveys administered to participants and nonparticipants in the Carolina Breast Cancer Study (CBCS). Surveys and structured interviews were administered to determine women's concerns regarding participation in research studies, access to health care, and beliefs regarding causes of breast cancer. Survey results showed the highest concern for the growing number of women diagnosed with breast cancer in North Carolina and potential environmental agents that may cause breast cancer. Negative responses were noted for time constraints related to participation and lack of familiarity with epidemiologic research; another concern noted was the lack of centralized information regarding breast cancer treatment. These issues were addressed by (1) developing a web site that provided background information about the CBCS, summaries of published study results, and information about the etiology of breast cancer; and (2) creating a statewide, comprehensive breast cancer resource directory for women who need information about breast cancer diagnosis, treatment, and support. These two projects were carried out in collaboration with breast cancer advocates, and demonstrate the important role that advocates can play in making epidemiologic research more responsive to the needs of communities. PMID- 11921177 TI - Breast cancer prevention through the eyes of a survivor. PMID- 11921176 TI - The partnership between breast cancer advocates and scientists. AB - The National Breast Cancer Coalition (NBCC) is a grassroots organization that represents breast cancer advocates and is committed to eradicating breast cancer. NBCC defines a breast cancer advocate as someone who has been personally affected by the disease (e.g., a breast cancer survivor, family member, or friend), represents a constituency, and is motivated to join the fight against the disease. One of the organization's goals is to ensure that breast cancer advocates have a seat at the table when decisions are made about breast cancer research and policy. To accomplish this goal, NBCC educates advocates so that they can participate in and make meaningful contributions to legislative, scientific, and regulatory decision-making bodies. In addition to creating educational opportunities for advocates, NBCC has spearheaded several initiatives designed to directly increase the quality and quantity of breast cancer research. NBCC has also played a major role in making funding available to breast cancer researchers. Two of NBCC's most notable programs include Project LEAD, an intensive science-training course for breast cancer advocates, and the Environmental Initiative, a collection of activities designed to improve research into the relationship between breast cancer and the environment. Breast cancer advocates trained by NBCC have partnered with the scientific community and individual scientists to improve the peer review, design, and implementation of breast cancer research. PMID- 11921178 TI - Human exposure to heterocyclic amine food mutagens/carcinogens: relevance to breast cancer. AB - Heterocyclic amines produced from overcooked foods are extremely mutagenic in numerous in vitro and in vivo test systems. One of these mutagens, 2-amino-1 methyl-6-phenylimidazo[4,5-b]pyridine (PhIP), induces breast tumors in rats and has been implicated in dietary epidemiology studies as raising the risk of breast cancer in humans. Efforts in our laboratory and others have centered on defining the exposure to PhIP and other dietary mutagens derived from cooked food. We accomplish this by analyzing the foods with a series of solid-phase extractions and HPLC. We have developed an LC/MS/MS method to analyze the four major human PhIP metabolites (sulfates and glucuronides) following a single meal containing 27 microg of cooking-produced PhIP in 200 g of grilled meat. Although the intake of PhIP was similar for each of eight women, the total amount excreted in the urine and the metabolite profiles differed among the subjects. It appears that adsorption (digestion) from the meat matrix, other foods in the diet, and genetic differences in metabolism may contribute to the variation. The four major metabolites that can be routinely assayed in the urine are N(2)-OH-PhIP-N(2) glucuronide, PhIP-N(2)-glucuronide, 4'-PhIP-glucuronide, and N(2)-OH-PhIP-N3 glucuronide. This work is suited to investigate individual exposure and risk, especially for breast cancer, from these potent dietary mutagens. PMID- 11921179 TI - Tobacco smoke carcinogens and breast cancer. AB - Cigarette smoking is an established cause of a variety of cancer types, but its role in breast cancer etiology is not clear. In this report, the potential role of cigarette smoke carcinogens as causes of human breast cancer is evaluated. Of over 60 known carcinogens in tobacco smoke, several are known to induce mammary tumors in laboratory animals: benzo[a]pyrene (B[a]P), dibenzo[a,l]pyrene (DB[a,l]P), 2-toluidine, 4-aminobiphenyl, 2-amino-3-methylimidazo[4,5-f]quinoline (IQ), 2-amino-1-methyl-6-phenylimidazo[4,5-b]pyridine (PhIP), 1,3-butadiene, isoprene, nitromethane, ethylene oxide, and benzene. Studies in humans demonstrate that tobacco constituents can reach breast tissue. The uptake and metabolic activation of mammary carcinogens such as polycyclic aromatic hydrocarbons (PAHs) and 4-aminobiphenyl are frequently higher in smokers than in nonsmokers. Although it is likely that specific mammary carcinogens in tobacco smoke can reach breast tissue, evidence is lacking at the present time. Some PAHs present in cigarette smoke can be metabolized to sterically hindered diol epoxides, which are potent mammary carcinogens. Thus, compounds such as benzo[c]phenanthrene (B[c]P), not classically considered to be a strong carcinogen in rodents, could nevertheless be metabolized in humans to diol epoxides carcinogenic to the breast. Collectively, the link between smoking and breast cancer is plausible but has been difficult to establish, probably because of the low carcinogen dose. PMID- 11921180 TI - Lavage and nipple aspiration of breast ductal fluids: a source of biomarkers for environmental mutagenesis. AB - The acquisition of breast ductal fluid by nipple aspiration and ductal lavage are simple noninvasive procedures to sample breast tissue. Nipple aspiration fluid (NAF) obtained with gentle suction and a simple syringe-adapted apparatus may evaluate the secretory components that bathe the ductal epithelial cells. Evaluations have included the quantification of soluble markers (carcinoembryonic antigen and prostatic-specific antigen), DNA amplification, protein gel electrophoresis, and mutagenesis assays. It has been suggested that environmental mutagens in the breast ductal system may contribute to carcinogenesis. The feasibility of mutagenesis assays on NAF has been limited by the small size of the samples obtained. Three small clinical studies detected mutagens in 6-14% of the samples using the Salmonella Ames assay. Ductal lavage collects more of a cellular aspirate from the ductal system utilizing a microcatheter. Early studies on ductal lavage fluid have included cytology and methylation-specific PCR. Ductal lavage in a high-risk group has identified cellular atypia in 21% of those sampled. Samples obtained through the nipple, by aspiration or lavage, are the proteinaceous secretions from the ductal system and ductal epithelial cells. The fluid represents the cellular events and the dynamic secretory process of the breast and may include potential initiators of the carcinogenesis process in the cellular microenvironment. Fluid obtained by ductal lavage may allow for more detailed studies of the role of mutagens in breast cancer. PMID- 11921181 TI - Evidence for the presence of mutagenic arylamines in human breast milk and DNA adducts in exfoliated breast ductal epithelial cells. AB - Aromatic and heterocyclic amines are ubiquitous environmental mutagens present in combustion emissions, fried meats, and tobacco smoke, and are suspect human mammary carcinogens. To determine the presence of arylamines in breast tissue and fluid, we examined exfoliated breast ductal epithelial cells for DNA adducts and matched human milk samples for mutagenicity. Breast milk was obtained from 50 women who were 4-6 weeks postpartum, and exfoliated epithelial-cell DNA was evaluated for bulky, nonpolar DNA adducts by (32)P-postlabeling and thin-layer chromatography. Milk was processed by acid hydrolysis, and the extracted organics were examined in the standard plate-incorporation Ames Salmonella assay using primarily strain YG1024, which detects frameshift mutations and overexpresses aryl amine N-acetyltransferase. DNA adducts were identified in 66% of the specimens, and bulky adducts migrated in a pattern similar to that of 4 aminobiphenyl standards. The distribution of adducts did not vary by NAT2 genotype status. Of whole milk samples, 88% (22/25) had mutagenic activity. Among the samples for which we had both DNA adduct and mutagenicity data, 58% (14/19) of the samples with adducts were also mutagenic, and 85% (11/13) of the mutagenic samples had adducts. Quantitatively, no correlation was observed between the levels of adducts and the levels of mutagenicity. Separation of the milk showed that mutagenic activity was found in 69% of skimmed milk samples but in only 29% of the corresponding milk fat samples, suggesting that the breast milk mutagens were moderately polar molecules. Chemical fractionation showed that mutagenic activity was found in 67% (4/6) of the basic fractions but in only 33% (2/6) of acidic samples, indicating that the mutagens were primarily basic compounds, such as arylamines. Although pilot in nature, this study corroborates previous findings of significant levels of DNA adducts in breast tissue and mutagenicity in human breast milk and indicates that breast milk mutagens may be moderately polar basic compounds, such as arylamines. PMID- 11921182 TI - Mutagens in human breast lipid and milk: the search for environmental agents that initiate breast cancer. AB - Epidemiological studies indicate the involvement of environmental factors in the etiology of breast cancer, but have not provided clear indications of the nature of the agents responsible. Several environmental carcinogens are known to induce mammary tumors in rodents, and the abundance of adipose tissue in the human breast suggests that the epithelial cells, from which breast tumors commonly arise, could be exposed to lipid-soluble carcinogens sequestered by the adipose tissue. In this report we review our studies in which we have examined human mammary lipid, obtained from elective reduction mammoplasties from healthy donors, and human milk from healthy mothers, for the presence of components with genotoxic activity in several in vitro assays. A significant proportion of lipid extracts induced mutations in bacteria and micronuclei in mammalian cells. They also caused DNA damage, detected as single-strand breaks in the alkaline single cell gel electrophoresis (comet) assay, in both the MCL-5 cell line and in primary cultures of human mammary epithelial cells. Genotoxic activity was also found in a significant proportion of extracts of human breast milk. Viable cells recovered from milk samples showed evidence of DNA damage and were susceptible to comet formation by genotoxic agents in vitro. Genotoxic activity was found to be less prevalent in milk samples from countries of lower breast cancer incidence (the Far East) compared with that in samples from the UK. The agents responsible for the activity in milk appear to be moderately polar lipophilic compounds and of low molecular weight. Identification of these agents and their sources may hold clues to the origins of breast cancer. PMID- 11921183 TI - Identification of mammary carcinogens in rodent bioassays. AB - Results from chemical carcinogenesis studies in rodents are useful to identify substances in our environment that may contribute to cancer development. The National Toxicology Program (NTP) was established in 1978 to coordinate research and testing of potential human carcinogens and to publish the Report on Carcinogens, which lists human carcinogens. The results for over 500 chemicals tested in the NTP 2-year bioassays have been published in Technical Reports and include data for chemical, agent, or complex mixture exposures. The bioassays have identified 42 chemicals that induce tumors in the rodent mammary gland. The physical and chemical characteristics of the carcinogens vary, but epoxides (including chemicals metabolized to epoxides) and nitro-containing compounds are well represented. The 9th Report on Carcinogens, issued in 2000, lists 21 of the 42 chemicals as human carcinogens including benzene, ethylene oxide, 1,3 butadiene, isoprene, chloroprene, C.I. basic red 9, and C.I. acid red 114. Ethylene oxide was associated with increased breast cancer risk in an epidemiologic study, whereas other listed chemicals, for which human data are available, display different target organ specificity. Bioassays other than those conducted by the NTP also provide information about rodent mammary gland carcinogens. Several carcinogen exposures are associated with breast tumor induction in both humans and rodents including radiation, diethylstilbestrol, and estrogens. These studies demonstrate that route, timing and frequency of exposure, and genetic factors contribute to the overall susceptibility to breast cancer development. More information is needed on the effects of chemicals to which humans are exposed and the manner by which they influence breast cancer risks. PMID- 11921184 TI - Studies on mammary carcinogenesis induced by a heterocyclic amine, 2-amino-1 methyl-6-phenylimidazo[4,5-b]pyridine, in mice and rats. AB - Ten heterocyclic amines (HCAs) that are produced by heating amino acids, proteins, or proteinaceous food such as fish and meat were examined for carcinogenicity in rats and mice. Three of them, 2-amino-3-methylimidazo[4,5 f]quinoline (IQ), 2-amino-3,4-dimethylimidazo[4,5-f]quinoline (MeIQ), and 2-amino 1-methyl-6-phenylimidazo[4,5-b]pyridine (PhIP), have been shown to induce mammary cancer in female F344 and/or SD rats, but none of the HCAs induced mammary cancer in CDF(1) mice. This report reviews our recent studies on mammary carcinogenesis of PhIP in various strains of mice and on the roles of genomic instability in the rat mammary carcinogenesis of PhIP. We demonstrated that the survival time from mammary adenocarcinomas was shorter in PhIP-treated BALB/c mice than that in the untreated control, and with a significantly higher incidence in the C.B-17 strain of mice compared with that of the control. To clarify mechanisms of mammary carcinogenesis, we examined genomic instability in rat mammary cancer induced by PhIP. Mammary cancers were induced in F344 x SD F(1) rats harboring the lacI transgene, and two cell lines were established from two adenocarcinomas. They showed a greater than 10-fold higher frequency of spontaneous mutations than that of the primary culture of normal mammary epithelial cells, in the lacI transgene and the hprt endogenous gene during cell replication. Nucleotide sequencing revealed that almost all types of mutations were increased, with a remarkable increase of A:T --> C:G mutation. This genomic instability was not attributed either to alterations of mismatch-repair enzymes or to p53. These mutational characteristics were also observed in the original tumors. Single-nucleotide instability (SNI) might be implicated in the mammary cancer induced by PhIP. PMID- 11921185 TI - Mammary gland carcinogenesis by food-derived heterocyclic amines: metabolism and additional factors influencing carcinogenesis by 2-amino-1-methyl-6 phenylimidazo[4,5-b]pyridine (PhIP). AB - The heterocyclic amines (HCAs) are a family of mutagenic/carcinogenic compounds found in cooked meats. Several HCAs are mammary gland carcinogens in rats. Of these compounds, 2-amino-1-methyl-6-phenylimidazo[4,5-b]pyridine (PhIP) is the major one present in the human diet. This report reviews the studies on rat mammary gland carcinogenesis by HCAs; discusses what is currently known regarding mechanisms of mammary gland carcinogenesis of PhIP, especially the significance of metabolic processing; and further highlights the evidence for the possible role of PhIP in human breast cancer. PMID- 11921186 TI - Tumor formation in Brca1 conditional mutant mice. AB - BRCA1 is the first breast cancer-associated gene, whose mutation predisposes women to breast and ovarian cancers. Targeted mutations of Brca1 in the mouse result in embryonic lethality primarily attributed to cellular proliferation defects, raising questions about the mechanisms by which Brca1 represses tumor formation. To overcome the early lethality, we engineered Brca1 by flanking its exon 11 with loxP sites. We showed that deletion of the exon by EIIA-Cre, which expresses Cre in the germline, causes p53-dependent lethality at late gestation. On the other hand, MMTV-Cre, which expresses Cre in mammary epithelium, resulted in tumorigenesis at low frequency after a long latency, accompanied by increased epithelial cell apoptosis and abnormal ductal development. Mammary tumor formation was significantly accelerated in a p53(+/-) genetic background; however, it still appeared in a stochastic fashion, suggesting the involvement of additional factors. Notably, the tumors were highly diverse in histopathology and displayed extensive genetic/molecular alterations, including overexpression of ErbB2, c-Myc, p27, and Cyclin D1, and downregulation of p16 in the majority of tumors. This observation suggests roles for these proteins in Brca1-associated tumorigenesis. PMID- 11921187 TI - Environmental carcinogens and p53 tumor-suppressor gene interactions in a transgenic mouse model for mammary carcinogenesis. AB - Mouse mammary tumorigenesis is greatly influenced by a variety of exogenous agents, such as MMTV, chemical carcinogens (i.e., polycyclic aromatic hydrocarbons), and radiation, as well as by endogenous/physiological factors, such as steroid hormones, tumor-suppressor genes (i.e., Brca1/2, p53), and gene products of modifier genes. In the mouse model, the most frequently used chemical carcinogen has been 7,12-dimethylbenz[a]anthracene (DMBA), which activates the Ha ras gene but does not alter the p53 tumor-suppressor gene. However, on an existing background of p53 gene alteration, low doses of DMBA are strongly cocarcinogenic. Using a transgenic model system, in which the p53 gene was deleted in the mammary gland, we examined the carcinogenic effects of a variety of external agents and internal factors given at either low doses or physiological doses. These agents/factors included DMBA, gamma-radiation, Brca2 heterozygosity, and steroid hormones. All agents/factors increased the tumorigenic response of the p53 null mammary cells, even under conditions where no tumorigenic response was observed in the p53 wildtype mammary cell. The strongest cocarcinogenic effect was observed with the steroid hormone progesterone. The majority of tumors were highly aneuploid and composed of nuclear igh-grade cells. The mechanism for the aneuploidy and secondary events associated with high tumorigenicity were examined using array technology. These results demonstrate that, on a background of underlying genetic instability, very low doses of environmental mutagens and mitogens can produce strong cocarcinogenic effects. PMID- 11921188 TI - Carcinogen-DNA adducts in human breast epithelial cells. AB - Diet and environmental exposures are often regarded as significant etiologic factors in human breast cancer. Chemicals that may be involved in these exposures include heterocyclic amines, aromatic amines, and polycyclic aromatic hydrocarbons, which also serve as strong mammary carcinogens in different animal models. In this study, we chose to quantify the major DNA adducts derived from one member of each of these classes of carcinogens, that is, 2-amino-1-methyl-6 phenylimidazo[4,5-b]pyridine (PhIP), 4-aminobiphenyl (ABP), and benzo[a]pyrene (B[a]P), respectively, in DNA isolated from exfoliated ductal epithelial cells in human breast milk. Milk was collected from healthy, nonsmoking mothers. The isolated DNA was digested to 3' nucleotides and subjected to (32)P-postlabeling. Adduct enrichment was achieved using Oasis Sep-Paks and the analyses were conducted by HPLC using radiometric detection. Critical to the analyses were the syntheses of bis(phosphate) standards for the C8-dG adducts of PhIP and ABP, and the N(2)-dG adduct of B[a]P, which were added to each reaction as UV markers. Of the 64 samples analyzed, adducts were found in 31 samples. Thirty samples contained detectable levels of PhIP adducts, with a mean value of 4.7 adducts/10(7) nucleotides; 18 were positive for ABP adducts with a mean value of 4.7 adducts/10(7) nucleotides; and 13 were found to contain B[a]P adducts with a mean level of 1.9 adducts/10(7) nucleotides. These data indicate that women are exposed to several classes of dietary and environmental carcinogens and that these carcinogens react with DNA in breast ductal epithelial cells, the cells from which most breast cancers arise. PMID- 11921189 TI - Characterization of a major aromatic DNA adduct detected in human breast tissues. AB - A bulky DNA adduct (Spot 1) was previously detected in normal adjacent breast tissues of 41% (36/87) of women with breast cancer and in none (0/29) of the noncancer controls by (32)P-postlabeling. To characterize this adduct, it was chromatographically compared with DNA adduct profiles generated in several in vitro and in vivo experimental systems. First, MCF-7 cells were exposed to a number of chemical carcinogens, that is, benzo[a]pyrene (B[a]P), 4-OH-B[a]P, 9-OH B[a]P, 11-OH-B[a]P, B[a]P-trans-4,5-dihydrodiol, 1-nitropyrene, 6-nitrochrysene, dibenzo[a,l]pyrene, benzo[c]phenanthrene, dibenzo[a,h]anthracene, 3 methylcholanthrene, and 2-amino-1-methyl-6-phenylimidazo[4,5-b]pyridine. Spot 1 was detected as a minor adduct in cells treated with B[a]P but not other compounds. Second, to determine whether Spot 1 is derived from lipid peroxidation products or estrogen metabolites, it was compared with adduct profiles of cells or DNAs exposed to 17beta-estradiol, 4-hydroxy estradiol, 4-hydroxynonenal, or oxidized oat oil. Spot 1 was not detectable in these samples. In addition, Spot 1 did not comigrate with the 1,N(2)-ethenodeoxyguanosine adduct standard. Third, to explore the mechanism of Spot 1 formation, it was compared with adduct profiles detected in DNA or mononucleotides reacted with BPDE, 1-OH-7,8-dihydrodiol of B[a]P, and 3-OH-7,8-dihydrodiol of B[a]P as well as in rats orally treated with B[a]P. Spot 1 comigrated with a minor adduct in BPDE-treated DNA during anion exchange rechromatography but these two adducts were separated by partition chromatography. Spot 1 also behaved in a manner that was very similar to that of the polar B[a]P adducts detected in rat liver, but the two adducts were separated by HPLC. Fourth, Spot 1 was compared with CD1 mice exposed to 7H-benzo[c]fluorene (B[c]F). Spot 1 from some patients comigrated with a major adduct induced by B[c]F. Finally, we found that the presence of Spot 1 in human breast tissues was not related to smoking status but, rather, with CYP1A1 MspI polymorphism. The CYP1A1 mutant carriers had a significantly higher frequency of this adduct than did the wild-type genotypes. Furthermore, individuals with Spot 1 had a significantly higher staining intensity for BPDE-PAH adducts in their tissue sections than those without it. These results demonstrate that this major bulky DNA adduct detected in human breast tissues is related to PAH exposure. PMID- 11921190 TI - Molecular epidemiologic studies of polycyclic aromatic hydrocarbon-DNA adducts and breast cancer. AB - We review our studies on the role of polycyclic aromatic hydrocarbon (PAH)-DNA adducts in breast cancer. Additionally we report on analyses of the reliability of the scoring procedures used with immunohistochemical assay for PAH-DNA adducts and of potential bias arising from the use of benign breast disease (BBD) controls. We conducted a case-control study utilizing two control groups: BBD controls who donated tissue and blood samples, and healthy controls who donated blood samples. In comparisons of tumor tissue from cases and benign tissue from BBD controls, increasing adduct levels were significantly associated with case control status [odds ratio (OR) = 2.40, 95% confidence interval (CI) 1.18-4.92], whereas in comparisons of nontumor tissue from cases and benign tissue from BBD controls the association was nonsignificant (OR = 1.97, 95% CI 0.94-4.17). We also show among cases, but not among BBD controls, that the GSTM1 null genotype is associated with increased adduct levels in breast tissue. Our reliability study found the scoring procedures used with the immunohistochemical assay to have high reliability, 0.93 in nontumor, 0.82 in tumor, and 0.74 in benign tissues. However, we found that the technician significantly contributed to the total variability of a series of data. Finally, we did not find a consistent bias to the null associated with the use of BBD controls; however, BBD controls may overestimate the prevalence of family history of breast cancer compared to that of healthy controls (18% vs.14%). We hypothesize that the higher prevalence results from a referral bias and discuss how this may influence our results. PMID- 11921191 TI - Genetic regulation of ionizing radiation sensitivity and breast cancer risk. AB - Genetic variability in DNA repair may contribute to hypersensitivity to ionizing radiation (IR) and susceptibility to breast cancer. We used samples collected from a clinic-based breast cancer case-control study to test the working hypothesis that amino acid substitution variants of DNA repair genes may contribute to prolonged cell-cycle delay following IR and breast cancer risk. Fluorescence-activated cell sorter (FACS) analysis was used to measure cell-cycle delay. PCR-restriction fragment length polymorphism (RFLP) assays were used to determine four genotypes of three DNA repair genes: XRCC1, 194 Arg/Trp and 399 Arg/Gln; XRCC3, 241 Thr/Met; and APE1, 148 Asp/Glu. The data showed that breast cancer patients had a significantly higher delay index than that of controls (P < 0.001); the means +/- SD for cases and controls were 36.0 +/- 13.1 (n = 118) and 31.4 +/- 11.5 (n = 225), respectively. There was a significant dose-response relationship between delay index, categorized into quartiles, and an increasing risk of breast cancer (crude odds ratios: 1.00, 1.00, 1.27, and 2.46, respectively; P(trend) = 0.002). In controls, prolonged cell-cycle delay was significantly associated with the number of variant alleles in APE1 Asp148Glu and XRCC1 Arg399Gln genotypes (P(trend) = 0.001). Although larger studies are needed to validate the results, our data suggest that an inherited hypersensitivity to IR may contribute to human breast carcinogenesis. PMID- 11921192 TI - p53 as a mutagen test in breast cancer. AB - The p53 gene is mutated in about half of all tumors. The p53 gene can be used as a "mutagen test," that is, the relative frequencies of the different types of mutation can be used as an epidemiological tool to explore the contribution of exogenous mutagens vs. endogenous processes in particular cancers. p53 has been used as a mutagen test in breast cancer. Surprisingly, the pattern of p53 mutations differs among 15 geographically and ethnically diverse populations. In contrast, mutation patterns in the human factor IX gene are similar in geographically and ethnically diverse populations. Diverse p53 mutation patterns in breast cancer are consistent with a significant contribution by a diversity of exogenous mutagens. Breast tissue may be uniquely sensitive to lipophilic mutagens because of its unique architecture, characterized by tiny islands of cancer-prone mammary epithelial cells surrounded by a sea of adipocytes. Mammary epithelial cells may be differentially susceptible to released lipophilic mutagens preferentially concentrated in adjacent adipocytes and originating in the diet. To test this hypothesis, we developed a method for measuring mutation load from ethanol-fixed, paraffin-embedded human tissues immunohistochemically stained with anti-p53 antibodies. Single cells staining positively for p53 overabundance are microdissected and the gene is sequenced. It is possible to identify individuals with a high mutation load in normal breast tissue and who are presumably at increased risk for breast cancer. In addition, analysis of the p53 gene with appropriate mutation detection methodology markedly improves the prediction of early recurrence, treatment failure, and death in breast cancer patients. Mutagen tests and mutation load measurements are useful tools to identify the role of mutagens in breast cancer. PMID- 11921193 TI - Inherited predisposition and breast cancer: modifiers of BRCA1/2-associated breast cancer risk. AB - Germline mutations in the BRCA1 and BRCA2 (BRCA1/2) genes explain a substantial proportion of hereditary breast and ovarian cancer. Women who have inherited a mutation in one of these genes are at increased risk to develop breast and/or ovarian cancer, although there is variability in the manifestation of tumors by age and site. This variability may be explained, in part, by the BRCA1/2 mutation type or location. However, it is also possible that risk-modifying factors exist that explain interindividual variability in cancer risk. These factors include genes at other loci and endogenous or exogenous exposures. A more complete understanding of factors that modify cancer risk in BRCA1 and BRCA2 mutation carriers may help to refine estimates of cancer risk. A number of exposures, including reproductive history and exogenous hormone use, have been implicated as BRCA1/2-associated cancer risk modifiers. Similarly, genes involved in hormone metabolism, including the AIB1 and AR genes, have been linked with altered breast cancer risk. Therefore, although germline BRCA1/2 mutations raise a woman's breast and ovarian cancer risk, other factors may interact with BRCA1/2 mutations to modulate this risk. PMID- 11921195 TI - 2-Amino-1-methyl-6-phenylimidazo[4,5-b]pyridine (PhIP)-induced mutagenesis in cultured Big Blue rat mammary epithelial and fibroblast cells. AB - Epithelial cells are the primary site of carcinogenesis in most tissues, including the mammary gland. As an alternative to the study of mutation induction in whole tissues in vivo, we have established Big Blue transgenic rat cell lines from the mammary epithelium (BBR/ME) and the mammary stroma (BBR/MFib), to permit a comparison of their mutagenic responses to carcinogens. We previously demonstrated their responsiveness to the alkylating agent N-ethyl-N-nitrosourea (ENU) (McDiarmid H et al. [2001]: Mutat Res 497:39-47). Here, we examined the responses of cultured epithelial and stromal cells to the protein pyrolysis product and mammary carcinogen 2-amino-1-methyl-6-phenylimidazo[4,5-b]pyridine (PhIP). Rat hepatic S9 was used as a source of bioactivation enzymes. Mutant induction (cII locus) and clonogenic survival were measured as a function of PhIP concentration. PhIP mutagenicity was observed in the fibroblast cells, but the greater toxicity of PhIP to the epithelial cells prevented a definitive evaluation of mutagenicity. Since PhIP may be detoxified by conjugation with glutathione, we measured glutathione levels and glutathione-S-transferase expression and activities in both cell lines. The epithelial cells had higher glutathione-S-transferase enzyme activity and protein expression than did the fibroblast cell line. Because the epithelial cells were more sensitive to toxicity, glutathione conjugation evidently plays only a minor role in PhIP toxicity and mutagenicity in our cell lines. PMID- 11921194 TI - Epigenetics of breast cancer: polycyclic aromatic hydrocarbons as risk factors. AB - In the absence of a causal relationship between the incidence of sporadic breast cancer and occurrence of mutations in breast cancer susceptibility genes, efforts directed to investigating the contribution of environmental xenobiotics in the etiology of sporadic mammary neoplasia are warranted. Polycyclic aromatic hydrocarbons (PAHs) are ubiquitous pollutants, which have been shown to induce DNA damage and disrupt cell cycle progression. In this report we discuss published data pointing to PAHs as a risk factor in carcinogenesis, and present findings generated in our laboratory suggesting that the mammary tumorigenicity of PAHs may be attributable, at least in part, to disruption of BRCA-1 expression by reactive PAH-metabolites. We report that benzo[a]pyrene (B[a]P), selected as a prototype PAH, disrupts BRCA-1 transcription in estrogen receptor (ER)-positive but not ER-negative breast cancer cells. The reduced potential for BRCA-1 expression in B[a]P-treated cells coincides with disruption of cell cycle kinetics and accumulation of p53. These effects are counteracted by the AhR antagonist alpha-naphthoflavone (ANF), and in breast cancer cells expressing mutant p53 or the E6 human papilloma virus protein. We suggest that exposure to PAHs may be a predisposing factor in the etiology of sporadic breast cancer by disrupting the expression of BRCA-1. PMID- 11921196 TI - Neoplastic transformation of human breast epithelial cells by estrogens and chemical carcinogens. AB - Sporadic breast cancer, the most common cancer diagnosed in American and Northern European women, is gradually increasing in incidence in most Western countries. Prevention would be the most efficient way of eradicating this disease. This goal, however, cannot be accomplished until the specific agent(s) or mechanisms that initiate the neoplastic process are identified. Experimental studies have demonstrated that mammary cancer is a hormone-dependent multistep process that can be induced by a variety of compounds and mechanisms, that is, hormones, chemicals, radiation, and viruses, in addition to or in combination with genetic factors. Although estrogens have been shown to play a central role in breast cancer development, their carcinogenicity on human breast epithelial cells (HBECs) has not yet been clearly demonstrated. Breast cancer initiates in the undifferentiated lobules type 1, which are composed of three cell types: highly proliferating cells that are estrogen-receptor negative (ER-), nonproliferating cells that are ER positive (ER+), and very few (<1%) ER+ cells that proliferate. Interestingly, endogenous 17beta-estradiol (E(2)) is metabolized by the cytochrome P450 enzyme isoforms CYP1A1 and CYP1B1, which also activate benzo[a]pyrene (B[a]P), a carcinogen contained in cigarette smoke. We postulate that if estrogens are carcinogenic in HBECs, they should induce the same transformation phenotypes induced by chemical carcinogens and ultimately genomic changes observed in spontaneously developing primary breast cancers. To test this hypothesis we compared the transforming potential of E(2) on the HBEC MCF-10F with that of B[a]P. Both E(2) and B[a]P induced anchorage-independent growth, colony formation in agar methocel, and loss of ductulogenic capacity in collagen gel, all parameters indicative of cell transformation. In addition, the DNA of E(2)-transformed cells expressed LOH in chromosome 11 at 11q23.3, 11q24.2-q25, and LOH at 13q12-q13. B[a]P-induced cell transformation was also associated with LOH at 13q12-q13 and at 17p13.2. The relevance of these findings is highlighted by the observation that E(2)- and B[a]P-induced genomic alterations in the same loci found in ductal hyperplasia, ductal carcinoma in situ, and invasive ductal carcinoma of the breast. PMID- 11921197 TI - Selective estrogen receptor modulators (SERMs) and retinoids in breast cancer chemoprevention. AB - Tamoxifen has been shown to decrease the risk of invasive breast cancer by 49% and noninvasive breast cancer by 50%. Tamoxifen is also associated with a threefold increased risk of endometrial cancer. Raloxifene, a second-generation selective estrogen receptor modulator (SERM), has not been associated with endometrial cancer risk, and is currently under study in a large, multi institutional, randomized Study of Tamoxifen and Raloxifene (STAR) for breast cancer prevention in postmenopausal women. A pilot trial of raloxifene in premenopausal women to assess the safety, tolerability, effects on bone mineral density, mammographic density, and other biological endpoints is ongoing. The retinoids have been shown to decrease mammary tumors in rodent carcinogenesis models. The Italian trial of fenretinide (4-HPR) in women with stage I breast cancer randomized women to fenretinide or no intervention. This study did not show an overall effect of decreasing the risk of contralateral breast cancer. However, a protective effect was suggested in premenopausal women. It has been suggested that this effect may be related to insulin-like growth factor 1 (IGF 1), which has been shown to be modulated by fenretinide in premenopausal but not postmenopausal women. Pilot studies of SERMs alone and in combination with retinoids or other agents provide a model for testing the safety and tolerability, pharmacokinetics and pharmacodynamics, and biomarker modulation in high-risk women. These studies can provide information as to both the pathophysiology of carcinogenesis and the mechanism of action of chemopreventive agents, and help select agents and doses for testing in large randomized studies. PMID- 11921198 TI - Chemoprevention of heterocyclic amine-induced mammary carcinogenesis in rats. AB - 2-Amino-1-methyl-6-phenylimidazo[4,5-b]pyridine (PhIP) is one of the most prevalent carcinogenic heterocyclic amines in the environment, targeting the colon, prostate, pancreas, and mammary gland in rodents. Chemopreventive effects of synthetic and naturally occurring compounds on PhIP-induced rat mammary carcinogenesis were investigated in a series of experiments. In a PhIP feeding model, groups of 20-21 female F344 rats each, were treated with 0.02% PhIP alone or PhIP plus 0.5% 1-O-hexyl-2,3,5-trimethylhydroquinone (HTHQ), 1% green tea catechins, 1% alpha-tocopherol, 0.1% ellagic acid, or 1% chlorophyllin, each in the diet, or 0.1% caffeine in drinking water for 52 weeks. To assess the mechanism of HTHQ and caffeine inhibition of PhIP-induced carcinogenesis, effects of these compound on the in vitro metabolic activation of PhIP were examined in the presence of S9 mix. In the next series of experiments, the PhIP intragastric dose model was applied to allow separate investigation of the effects of chemicals during the initiation and postinitiation periods. In these experiments, female Sprague-Dawley rats were given eight intragastric doses of 100 mg/kg body weight during the first 4-8 weeks for initiation. Either during initiation or after initiation, or only after initiation, animals were treated with either corn or perilla oil at doses of 5 and 20%, conjugated fatty acid derived from safflower oil (CFA-S) or perilla oil (CFA-P) at a dose of 1%, arctiin at doses of 0.02 and 0.2% in the diet, or sodium nitrite (NaNO(2)) at a dose of 0.2% in drinking water. In the PhIP feeding model, administration of PhIP alone for 52 weeks induced adenocarcinomas in 40% of rats, but the incidence was remarkably reduced to 5% by the simultaneous treatment with 0.5% HTHQ, a strong lipophilic phenolic antioxidant, or to 10% by 0.1% caffeine. Administration of 1% chlorophyllin exerted similar, albeit weaker, effects. alpha-Tocopherol at a dose of 0.5% only reduced the multiplicity of carcinomas, and 1% green tea catechins only the mean size of mammary tumors. In a metabolic activation study of PhIP, HTHQ and caffeine clearly inhibited the formation of metabolites. In the PhIP gastric dose model, among the naturally occurring compounds examined, a plant lignan arctiin, perilla oil, which contains a large amount of n-6 alpha-linolenic acid, and CFA-S or CFA-P inhibited mammary tumor development, particularly in the postinitiation period, although a clear dose response was not observed. Treatment with 0.2% NaNO(2) in the initiation period was found to lower the volume of mammary tumors. The present results indicate that a number of compounds may be candidate chemopreventive agents against PhIP-induced mammary carcinogenesis, acting through different mechanisms and depending on the stage of carcinogenesis. PMID- 11921199 TI - Glial expression of small heat shock proteins following an excitotoxic lesion in the immature rat brain. AB - Heat shock proteins (HSPs) are chaperones induced under pathological conditions and involved in protein stabilization and cellular protection. In this study, we have evaluated the expression pattern of the glial cell-related HSP27, HSP32, and HSP47 following an excitotoxic lesion in the immature rat brain. Postnatal day 9 rats received an intracortical injection of N-methyl-D-aspartate and tissue was processed immunohistochemically for HSPs and double labeling using astroglial and microglial markers. HSP expression was quantified by image analysis. Excitotoxic damage caused primary cortical degeneration and secondary damage in the corresponding thalamus. In the injured cortex, reactive microglia/macrophages expressed HSP32 from 10 h until 14 days postlesion (PL), showing maximal levels at days 3-5. In parallel, most cortical reactive astrocytes showed expression of HSP47 from 10 h until 14 days PL and a population of them also displayed HSP27 labeling from 1 day PL. In addition, some cortical reactive astrocytes showed a temporary expression of HSP32 at day 1. In general, astroglial HSP expression in the cortex achieved maximal levels at days 3-5 PL. In the damaged thalamus, HSP32 was not significantly induced, but reactive astrocytes expressed HSP47 and some of them also HSP27. Thalamic astroglial HSP induction was transient, peaked at 5 days PL and reached basal levels by day 14. The injury-induced expression of HSP32, HSP27, and HSP47 in glial cells may contribute to glial cell protection and adaptation to damage, therefore playing an important role in the evolution of the glial response and the excitotoxic lesion outcome. HSP32 may provide antioxidant protective mechanisms to microglia/macrophages, whereas HSP47 could contribute to extracellular matrix remodeling and HSP27 may stabilize the astroglial cytoskeleton and participate in astroglial antioxidant mechanisms. PMID- 11921200 TI - Trisialoganglioside GT1b induces in vivo degeneration of nigral dopaminergic neurons: role of microglia. AB - We recently showed that trisialoganglioside (GT1b) induces cell death of dopaminergic neurons in rat mesencephalic cultures (Chung et al., Neuroreport 12:611-614, 2001). The present study examines the in vivo neurotoxic effects of GT1b on dopaminergic neurons in the substantia nigra (SN) of Sprague-Dawley rats. Seven days after GT1b injection into the SN, immunocytochemical staining of SN tissue revealed death of nigral neurons, including dopaminergic neurons. Additional immunostaining using OX-42 and OX-6 antibodies showed that GT1b activated microglia were present in the SN where degeneration of nigral neurons was found. Western blot analysis and double-labeled immunohistochemistry showed that inducible nitric oxide synthase (iNOS) was expressed in the SN, where its levels were maximal at 8 h post-GT1b injection, and that iNOS was localized exclusively within microglia. GT1b-induced loss of dopaminergic neurons in the SN was partially inhibited by N(G)-nitro-L-arginine methyl ester hydrochloride, an NOS inhibitor. Our results indicate that in vivo neurotoxicity of GT1b against nigral dopaminergic neurons is at least in part mediated by nitric oxide released from activated microglia. Because GT1b exists abundantly in central nervous system neuronal membranes, our data support the hypothesis that immune-mediated events triggered by endogenous compounds such as GT1b could contribute to the initiation and/or the progression of dopaminergic neuronal cell death that occurs in Parkinson's disease. PMID- 11921201 TI - Cellular localization and expression patterns of interleukin-10, interleukin-4, and their receptors in multiple sclerosis lesions. AB - Cytokines have been shown to play a crucial role in the pathogenesis of multiple sclerosis (MS). However, still limited data are available on the expression of anti-inflammatory cytokines within the central nervous system (CNS) during MS lesion development. Therefore, we have examined the expression of the anti inflammatory cytokines, interleukin-10 (IL-10) and IL-4, and their specific receptors, IL-10R and IL-4R, in postmortem human brain tissue obtained from MS patients. Specific patterns of protein localization and expression for both proteins could be observed within active and chronic MS lesions. Strongest IL-10 immunoreactivity was observed in reactive astrocytes within active demyelinating lesions and the hypercellular rim of chronic active MS lesions. Moreover, perivascular macrophages were immunoreactive for IL-10 in (chronic) active MS lesions. Most intense IL-4 immunoreactivity was detected in reactive fibrillary astrocytes within the hypocellular regions of chronic active and chronic inactive MS lesions. Strong immunoreactivity for IL-10R and IL-4R was detected on macrophages in both parenchymal and perivascular areas and on reactive astrocytes in active and chronic MS lesions. Our results indicate that IL-10 and IL-4 have an active role in CNS immune responses. The specific patterns of protein localization and protein expression for both IL-10 and IL-4 in MS lesions at different stages of development suggest that these anti-inflammatory cytokines and their receptors participate in processes leading to the formation of chronic MS lesions. PMID- 11921202 TI - TrkA immunoreactive astrocytes in dendritic fields of the hippocampal formation across estrous. AB - Neurotrophins are important modulators of structural synaptic plasticity. (Through trophic action (Jordan. J Neurobiol 40:434-445, 1999), astrocytes serve as permissive substrates to support axonal regrowth (Ridet et al. Trends Neurosci 20:570-571, 1997), and are involved in estrogen-induced synaptic structural plasticity (Garcia-Segura et al. Cell Mol Neurobiol 16:225-237, 1996). Previously, we reported that tyrosine kinase A receptor (TrkA) immunoreactivity was present both in presynaptic neuronal processes (axons and terminals) and in select astrocytes of the male rat hippocampal formation (Barker-Gibb et al. J Comp Neurol 430:182-199, 2001). We show that the number of TrkA-immunoreactive astrocytes in female rats fluctuates 16-fold across the estrous cycle in dendritic fields of the hippocampal formation, with the greatest number at estrus after the peak plasma estradiol concentration of proestrus. Few TrkA-labeled astrocytes were found in ovariectomized animals; after estrogen replacement, this number increased by 12-fold in the hippocampal formation, indicating estrogen mediated induction. Dual-labeling studies showed that TrkA-labeled astrocytes were also immunoreactive for vimentin, a protein expressed by reactive astrocytes. Ultrastructural analysis of the dentate gyrus molecular layer demonstrated that TrkA immunoreactive astrocytes are positioned primarily next to dendrites and unmyelinated axons. Because nerve growth factor (NGF) has been reported to stimulate astrocytes to function as substrates for axon growth (Kawaja and Gage. Neuron 7:1019-1030, 1991), these findings are consistent with the theory that TrkA immunoreactive astrocytes serve a role in structural plasticity, axon guidance, and synaptic regeneration across the estrous cycle in the hippocampal formation. PMID- 11921204 TI - Isolation of a glial-restricted tripotential cell line from embryonic spinal cord cultures. AB - Neuroepithelial stem cells (NEPs), glial-restricted precursors (GRPs), and neuron restricted precursors (NRPs) are present during early differentiation of the spinal cord and can be identified by cell surface markers. In this article, we describe the properties of GRP cells that have been immortalized using a regulatable v-myc retrovirus construct. Immortalized GRP cells can be maintained in an undifferentiated dividing state for long periods and can be induced to differentiate into two types of astrocytes and into oligodendrocytes in culture. A clonal cell line prepared from immortalized GRP cells, termed GRIP-1, was also shown to retain the properties of a glial-restricted tripotential precursor. Transplantation of green fluorescent protein (GFP)-labeled subclones of the immortalized cells into the adult CNS demonstrates that this cell line can also participate in the in vivo development of astrocytes and oligodendrocytes. Late passages of the immortalized cells undergo limited transdifferentiation into neurons as assessed by expression of multiple neuronal markers. The availability of a conditionally immortalized cell line obviates the difficulties of obtaining a large and homogeneous population of GRPs that can be used for studying the mechanism and signals for glial cell differentiation as well as their application in transplantation protocols. PMID- 11921203 TI - Differential gene expression in astrocytes from human normal and glaucomatous optic nerve head analyzed by cDNA microarray. AB - Recent advances in cDNA microarray technology have made it possible to analyze expression of several thousand genes at the same time. Using this technique, gene expression in human astrocytes cultured from glaucomatous and normal optic nerve heads (ONH) was compared. One hundred-fifty genes were differentially expressed more than 5-fold in glaucomatous cell cultures compared with normal. These genes are involved in a number of biological processes, including signal transduction, cell adhesion and proliferation, ECM synthesis, and degradation. Confirmation of differential gene expression was performed by quantitative RT-PCR. Western blots and immunohistochemistry demonstrated gene products in cell cultures or in human ONH tissues. Proliferation, adhesion and migration assays tested physiological responses suggested by differential gene expression. Our study suggests that cultured glaucomatous ONH astrocytes retain in culture many phenotypic characteristics that may be relevant to their role in the pathogenesis of glaucoma and, in general to reactive astrocytes in the CNS. Potential applications of these data include the identification and characterization of signaling pathways involved in astrocyte function, studies of the role of steroid metabolizing enzymes in the glaucomatous ONH, and further exploration of the role of selected identified genes in experimental animal and in vitro models of glaucoma. PMID- 11921205 TI - Mechanism of NMDA receptor contribution to axon-to-glia signaling in the crayfish medial giant nerve fiber. AB - Electrical stimulation of crayfish giant axons at high frequency activates group II metabotropic and NMDA glutamate receptors on adjacent glial cells via release of N-acetylaspartylglutamate and glutamate formed upon its hydrolysis. This produces a transient depolarization followed by a prolonged hyperpolarization of glial cells that involves nicotinic acetylcholine receptor activation. The hyperpolarization is nearly completely blocked by antagonists of metabotropic glutamate receptors but only slightly reduced by inhibition of NMDA receptors. We report that the NMDA-induced hyperpolarization of glial cells is reduced by decreased calcium in the solution bathing the giant nerve fiber, while removal of sodium ions or block of voltage-dependent calcium channels completely prevents the glial response to NMDA. Inhibition of nicotinic acetylcholine receptors or removal of extracellular Cl(-) converts the glial response from a hyperpolarization to a depolarization that is sensitive to NMDA receptor antagonist. We propose that NMDA receptor activation by glutamate, formed from extracellular N-acetylaspartylglutamate during nerve stimulation, contributes to glial hyperpolarization by increasing intracellular Ca(2+) via opening of voltage sensitive Ca(2+) channels. Based on our previous work, we propose further that the added Ca(2+) supplements that produced by N-acetylaspartylglutamate and glutamate acting on group II metabotropic glutamate receptors to cause an increased release of acetylcholine and a larger hyperpolarization. PMID- 11921206 TI - Oligodendrocytes from human donors differ in resistance to herpes simplex virus 1 (HSV-1). AB - Primary cultures of human oligodendrocytes (HOLs) were established from six different donors. Differences in resistance to infection with herpes simplex virus 1 (HSV-1) were determined between the primary cultures of HOL in tissue culture infective dose 50 (TCID(50)), indirect immunofluoresence (IF), and serial electron microscopy (EM) studies. Virus production at different multiplicities of infection (MOIs) indicated that differences in HSV-1 replication were statistically significant and MOI-dependent. Overall, virus yield from the HOL cultures infected at an MOI of 1 increased up to 6 days postinfection (PI); no additional enhancement occurred at 7 days PI. However, differences in the replication capacity of the six HOL cultures observed at 5 days PI persisted at 6 and 7 days PI. When taken together, the results of these investigations indicate that, similar to experimental animals, resistance to HSV-1 differs between primary cultures of HOL and is donor-dependent. The results also raise the possibility that similar to experimental animals, resistance to HSV-1, mediated at the level of HOL, may be genetically determined. Furthermore, permissive infections of primary cultures of HOL were established with HSV-1 over a wide range of MOIs, similar to results obtained with viral infection of primary murine oligodendrocytes, but neither latent nor abortive infections of HOL were induced in vitro, even at very low MOIs. Resistance to HSV-1, mediated by glial cells, is a nonimmune mechanism that may influence the development of acute CNS infection in man as well as individual susceptibility to this virus. PMID- 11921209 TI - Inorganic semiconductor nanowires: rational growth, assembly, and novel properties. AB - Rationally controlled growth of inorganic semiconductor nanowires is important for their applications in nanoscale electronics and photonics. In this article, we discuss the rational growth, physical properties, and integration of nanowires based on the results from the authors' laboratory. The composition, diameter, growth position, and orientation of the nanowires are controlled based on the vapor-solid-liquid (VLS) crystal growth mechanism. The thermal stability and optical properties of these semiconductor nanowires are investigated. Particularly, ZnO nanowires with well-defined end surfaces can function as room temperature ultraviolet nanolasers. In addition, a novel microfluidic-assisted nanowire integration (MANI) process was developed for the hierarchical assembly of nanowire building blocks into functional devices and systems. PMID- 11921210 TI - Three-coordinate [Cu(II)X(3)](-) (X=Cl, Br), trapped in a molecular crystal. AB - Mixtures of [Ph(3)PNPPh(3)](+)Cl(-) with CuBr(2) (or CuBr(2)+CuCl(2)) in ethanol/dichloromethane yield crystals containing three-coordinate copper(II) with mixed chloride and bromide ligands, namely [Ph(3)PNPPh(3)](+)[CuCl(0.9)Br(2.1)](-) (1) and [Ph(3)PNPPh(3)](+)[CuCl(2.4)Br(0.6)](-) (2). The trigonal-planar coordination of copper(II) is angularly distorted but unambiguous, as there is no other halide ligand within 6.7 A of the copper atom. Density functional theory (DFT) calculations on planar [CuClBr(2)](-) show that the energy surface for angle bending is very soft. Crystallisation in the presence of CH(3)CN yields [Ph(3)PNPPh(3)](+)[CuCl(0.7)Br(2.3)(NCCH(3))](-) (3), in which there is additional secondary coordination by NCCH(3) (Cu-N 2.44 A). DFT calculations of the potential energy surface for this secondary coordination show that it is remarkably flat (<3 kcal mol(-1) for a variation of Cu-N by 0.8 A). The crystal packing in 1, 2 and 3, which involves multiple phenyl embraces between [Ph(3)PNPPh(3)](+) ions and numerous C-H...Cl and C-H...Br motifs, is associated with intermolecular energies that are larger than the variations in intramolecular energies. For reference, the crystal structures of [Ph(3)PNPPh(3)(+)](2)[Cu(2)Cl(6)](2-) (4) and [Ph(3)PNPPh(3)(+)](2)[Cu(2)Br(6)](2 ) (5) are described. We conclude 1) that three-coordinate copper(II) with monatomic halide ligands, although uncommon, can be regarded as normal, 2) that steric control by ligands is not necessary to enforce three-coordination, 3) that a hydrophobic aryl environment stabilises [Cu(Cl/Br)(3)](-), and 4) that the energy change in the transition from three- to four-coordinate copper(II) is very small (ca 5 kcal mol(-1)). PMID- 11921211 TI - Stereodynamics of bond rotation in tertiary aromatic amides. AB - The degree to which the rotations about the C-N and Ar-CO bonds of aromatic amides occur in a concerted manner was investigated by a variety of NMR and kinetic techniques. Otherwise complex kinetic analyses were simplified by exploiting symmetry and asymmetry in the N-substituents of amides. In 2 unsubstituted 1-naphthamides bearing branched N-substituents, most conformational changes about the amide group were by correlated rotation, though uncorrelated Ar CO rotation also occurred to some extent. In 2-substituted 1-naphthamides, correlated rotation accounted for all of the Ar-CO rotations, though a significant amount of uncorrelated C-N rotation also occurred. Naphthamides bearing branched N-substituents thus turn out to be efficient molecular gears: Compound 12 showed almost no gear slippage. PMID- 11921212 TI - Mechanism of the forbidden [3s,5s]-sigmatropic shift: orbital symmetry influences stepwise mechanisms involving diradical intermediates. AB - The mechanisms of [3s,5s]-sigmatropic shifts of octa-1,3,7-triene and 7 methylenenona-1,3,8-triene have been elaborated using B3LYP and BPW91 density functional theory and CASPT2 methods. These orbital symmetry forbidden rearrangements are stepwise, involving diradical intermediates. A comparison with several [3,3]-sigmatropic shifts of substituted hexadienes and of [5,5] sigmatropic shifts that are allowed, but nevertheless follow stepwise paths, shows that the activation barrier for the disallowed [3,5] shift is significantly larger than that for the stepwise reactions that are orbital symmetry allowed. Cyclic diradicals that have an aromatic circuit of electrons including the two radical centers and conjugated pi or sigma bonds are stabilized as compared to cyclic diradicals with an antiaromatic circuit of electrons. This applies to the transition states leading to and from the diradicals and influences the activation energies of stepwise sigmatropic shifts. The magnitudes of these effects are small but will have a significant influence on the rates of competing processes. This series of calculations has been used to assess the relative capabilities of the two functionals. We find that BPW91 underestimates the endothermicity of diradical formation and the barrier to diradical formation whereas B3LYP overestimates these quantities. PMID- 11921213 TI - Synthesis of guanidinium-derived receptor libraries and screening for selective peptide receptors in water. AB - A library of "tweezer" receptors, incorporating a guanidinium "head group" and two peptide derived side arms has been prepared on the solid-phase using an orthogonally protected guanidinium scaffold 12. The library was screened with various tripeptide derivatives in an aqueous solvent system. A tweezer receptor 25 for the side chain protected tripeptide 19 was identified from the screening experiments. Receptor 25 was resynthesised and solution binding studies were carried out, which revealed that 25 binds to tripeptide 19 with K(a)=8.2 x 10(4) +/- 2.5 x 10(4) (15 % DMSO/H(2)O, pH 8.75) and with appreciable selectivity over the tripeptide enantiomer 22 and the side chain deprotected tripeptide 20. PMID- 11921214 TI - Coordination and oxidative addition at a low-coordinate rhodium(I) beta-diiminate centre. AB - The reaction of 14e [L(Me)Rh(coe)] (1; L(Me)[double bond]ArNC(Me)CHC(Me)NAr, Ar[double bond]2,6-Me(2)C(6)H(3); coe[double bond]cis-cyclooctene) with phenyl halides and thiophenes was studied to assess the competition between sigma coordination, arene pi coordination and oxidative addition of a C-X bond. Whereas oxidative addition of the C-Cl and C-Br bonds of chlorobenzene and bromobenzene to L(Me)Rh results in the dinuclear species [[L(Me)Rh(Ph)(micro-X)](2)] (X=Cl, Br), fluorobenzene yields the dinuclear inverse sandwich complex [[L(Me)Rh](2)(anti-micro-eta(4):eta(4)-PhF)]. Thiophene undergoes oxidative addition of the C-S bond to give a dinuclear product. The reaction of 1 with dibenzo[b,d]thiophene (dbt) in the ratio 1:2 resulted in the formation of the sigma complex [L(Me)Rh(eta(1)-(S)-dbt)(2)], which in solution dissociates into free dbt and a mixture of the mononuclear complex [L(Me)Rh(eta(4)-(1,2,3,4)-dbt)] and the dinuclear complex [[L(Me)Rh](2)(micro-eta(4)-(1,2,3,4):eta(4)-(6,7,8,9) dbt)]. The latter could be obtained selectively by the 2:1 reaction of 1 and dbt. Reaction of 1 with diethyl sulfide produces [L(Me)Rh(Et(2)S)(2)], which in the presence of hydrogen loses a diethyl sulfide ligand to give [L(Me)Rh(Et(2)S)(H(2))] and catalyses the hydrogenation of cyclooctene. PMID- 11921215 TI - A novel approach for the oxidative degradation of organic pollutants in aqueous solutions mediated by iron tetrasulfophthalocyanine under visible light radiation. AB - An efficient approach has been developed to decompose toxic organic pollutants. The photodegradation of Rhodamine B (RhB), salicylic acid, and Orange II was examined in the presence of iron tetrasulfophthalocyanine ([Fe(PcS)]) and H(2)O(2) under visible irradiation. It was found that under visible light irradiation, organic pollutants in the [Fe(PcS)]/H(2)O(2) system can be rapidly degraded, but the concentration of [Fe(PcS)] remains nearly unchanged, and this indicates that [Fe(PcS)] has a good catalytic character. EPR results and other experimental results suggest that the light-activated reaction process involves the formation and reaction of HO(.) radicals. On the basis of the experimental results, a possible reaction mechanism for the degradation of organic pollutants under visible light illumination in the aqueous [Fe(PcS)]/H(2)O(2) solutions is proposed. PMID- 11921216 TI - Dinuclear complexes with bis(benzenedithiolate) ligands. AB - As a part of a broader study directed towards helical coordination compounds with benzenedithiolate donors, we have synthesized the bis(benzenedithiol) ligands 1,2 bis(2,3-dimercaptobenzamido)ethane (H(4)-1) and 1,2-bis(2,3 dimercaptophenyl)ethane (H(4)-2). Both ligands form dinuclear complexes with Ni(II), Ni(III) and, after air-oxidation, Co(III) ions under equilibrium conditions. Complexes (NEt(4))(4)[Ni(II)(2)(1)(2)] (11 b), (NEt(4))(2)[Ni(III)(2)(1)(2)] (13), and Na(4)[Ni(II)(2)(2)(2)] (14) were characterized by X-ray diffraction. In all complexes, two square-planar [Ni(S(2)C(6)H(3)R)(2)] units are linked in a double-stranded fashion by the carbon backbone and they assume a coplanar arrangement in a stair-like manner. Cyclic voltammetric investigations show a strong dependence of the redox potential on the type of the ligand. The substitution of 1(4-) for 2(4-) on nickel (-785 mV for 11 b versus -1130 mV for 14, relative to ferrocene) affects the redox potential to a similar degree as the substitution of nickel for cobalt (-1160 mV for [Co(2)(1)(2)](2-)/[Co(2)(1)(2)](4-), relative to ferrocene). The redox waves display a markedly less reversible behavior for complexes with the shorter bridged ligand 2(4-) compared to those of 1(4-). PMID- 11921217 TI - The hetero-Diels-Alder addition of sulfur dioxide to 1-fluorobuta-1,3-dienes: the sofa conformations preferred by 6-fluorosultines (6-fluoro-3,6-dihydro-1,2 oxathiin-2-oxides) enjoy enthalpic and conformational Anomeric effects. AB - The reactivity of (E)- and (Z)-1-fluorobuta-1,3-diene ((E)- and (Z)-11), 2 fluorobutadiene (12), (E)- and (Z)-1-(fluoromethylidene)-2-methylidenecyclohexane ((E)- and (Z)-13) toward SO(2) has been explored and compared with that of (Z)- and (E)-1-(fluoromethylidene)-2-methylidene-3,4-dihydronaphthalene ((Z)-8 and (E) 8). In agreement with quantum calculations, 12 is unreactive toward SO(2) (no cycloaddition, only polymerization), whereas (E)-1-fluoro-1,3-dienes react more rapidly than their (Z)-isomers to give the corresponding 6-fluorosultines following the endo (Alder rule) mode of hetero-Diels-Alder addition. No sulfolene has been observed following the cheletropic mode of addition with the fluorodienes, in contrast to other substituted dienes. In agreement with the calculations, cis-2-fluoro-3,4-oxathiabenzobicyclo[4.4.0]dec-1(6),9-diene-4-oxide (cis-9, the sultine obtained by SO(2) addition to (Z)-8 under conditions of kinetic control) adopts a sofa conformation with the oxygen atom of the ring lying in the average plane of the four carbon atoms of its sultine moiety when it is in the crystalline state at -100 degrees C. A similar sofa conformation was found for its trans-isomer, trans-9, obtained by isomerization of cis-9 or by hetero-Diels-Alder addition of SO(2) to (E)-8. Experiments (equilibrium constant for hetero-Diels-Alder additions, bond lengths, and bond angles in crystalline fluorosultines cis-9 and trans-9) and high-level quantum calculations on cis- and trans-6-fluoro-3,6-dihydro-1,2-oxathiin-2-oxide (cis- and trans-20) confirm the existence of a stabilizing, enthalpic, anomeric (gem-disubstitution by sulfinyloxy and fluoro groups) effect, which is interpreted in terms of (lone pair) n(O1)-->sigma*(C-F) hyperconjugative interactions. This effect is strongest in the sofa conformers with a gauche arrangement of the sigma(O1,S2) and sigma(C6,F) bonds. The calculations suggest also that n(O1)-->sigma*(S2,O2'), pi*(S=O), and n(S2)-->sigma*(O1,C6) interactions intervene and affect the relative stability of the conformers (sofa, boat, pseudo-chair) found for 6 fluorosultines cis- and trans-20. PMID- 11921218 TI - Formation of water-in-CO(2) microemulsions with non-fluorous surfactant Ls-54 and solubilization of biomacromolecules. AB - The solubility of Ls-54 surfactant in supercritical CO(2) was determined. It was found that the surfactant was highly soluble in SC CO(2) and the water-in-CO(2) microemulsions could be formed, despite it being a non-fluorous and non-siloxane nonionic surfactant. The main reasons for the high solubility and formation of the microemulsions may be that the surfactant has four CO(2)-philic groups (propylene oxide) and five hydrophilic groups (ethylene oxide) and its molecular weight are relatively low. The results of this work provide useful information for designing CO(2)-soluble non-fluorous and non-siloxane surfactants. The phase behavior of the CO(2)/Ls-54/H(2)O system, solvatochromic probe study, and the UV spectrum of lysozyme proved the existence of water domains in the SC CO(2) microemulsions. The method of synchrotron radiation small-angle X-ray scattering was used to obtain the structural information on the Ls-54 based water-in-CO(2) reverse micelles. By using the Guinier plot (ln I(q) versus q (2)) on the data sets in a defined small q range (0.022-0.040 A(-1)), the radii of the reverse micelles were obtained at different pressures and molar ratio of water to surfactant, W(0), which were in the range of 20.4-25.2 A. PMID- 11921219 TI - Photodissociation and electronic spectroscopy of [Re(H)(CO)(3)(H-dab)] (H-dab=1,4 diaza-1,3-butadiene): quantum wavepacket dynamics based on ab initio potentials. AB - The photodissociation dynamics of [Re(H)(CO)(3)(H-dab)] (H-dab=1,4-diaza-1,3 butadiene) were studied by means of wavepacket propagations on CASSCF/MR-CCI potentials calculated for the electronic ground state and low-lying excited states as a function of two coordinates, q(a) and q(b), that correspond to the Re H bond homolysis and to the axial CO loss, respectively. The theoretical absorption spectrum is characterized by two bands, one intense peak centered at lambda=500 nm (21,000 cm(-1)) and one broad band centered at 310 nm (32,500 cm( 1)). The visible band was assigned to the low-lying metal-to-ligand charge transfer (MLCT) states with a main contribution of the a(1)A'-->c(1)A' transition corresponding to the 3d(xz)-->pi*(dab) excitation. The second band calculated in the UV energy domain was assigned to the d(1)A' (sigma(Mn-H)-->pi*(dab)) state corresponding to a sigma-bond-to-ligand charge-transfer (SBLCT) state. The photodissociation dynamics of the low-lying (1)MLCT and (3)SBLCT states following irradiation in the visible energy domain was simulated by wavepacket propagation on the two-dimensional diabatic potentials V(q(a), q(b)) coupled by the spin orbit. In contrast to what was found for the manganese analogue, the (1)MLCT state is nonreactive and a rather slow (beyond the ps time scale), nontotal and indirect homolysis of the Re-H bond occurs through (1)MLCT-->(3)SBLCT intersystem crossing. PMID- 11921220 TI - Crystallization-induced diastereoselection: asymmetric synthesis of substance P inhibitors. AB - A novel three-component condensation followed by a crystallization-induced asymmetric transformation is used to build this key substance P inhibitor intermediate in a short synthetic sequence. PMID- 11921221 TI - Alkali metal ion binding to amino acids versus their methyl esters: affinity trends and structural changes in the gas phase. AB - The relative alkali metal ion (M(+)) affinities (binding energies) between seventeen different amino acids (AA) and the corresponding methyl esters (AAOMe) were determined in the gas phase by the kinetic method based on the dissociation of AA-M(+)-AAOMe heterodimers (M=Li, Na, K, Cs). With the exception of proline, the Li(+), Na(+), and K(+) affinities of the other aliphatic amino acids increase in the order AAAAOMe is already observed for K(+). Proline binds more strongly than its methyl ester to all M(+) except Li(+). Ab initio calculations on the M(+) complexes of alanine, beta-aminoisobutyric acid, proline, glycine methyl ester, alanine methyl ester, and proline methyl ester show that their energetically most favorable complexes result from charge solvation, except for proline which forms salt bridges. The most stable mode of charge solvation depends on the ligand (AA or AAOMe) and, for AA, it gradually changes with metal ion size. Esters chelate all M(+) ions through the amine and carbonyl groups. Amino acids coordinate Li(+) and Na(+) ions through the amine and carbonyl groups as well, but K(+) and Cs(+) ions are coordinated by the O atoms of the carboxyl group. Upon consideration of these differences in favored binding geometries, the theoretically derived relative M(+) affinities between aliphatic AA and AAOMe are in good overall agreement with the above given experimental trends. The majority of side chain functionalized amino acids studied show experimentally the affinity order AAAAOMe. The latter ranking is attributed to salt bridge formation. PMID- 11921222 TI - Chemoenzymatic synthesis of a bifunctionalized cellohexaoside as a specific substrate for the sensitive assay of cellulase by fluorescence quenching. AB - A new bifunctionalized cellohexaose derivative was synthesized as a specific substrate for continuous assay of cellulases by resonance energy transfer. This cellohexaoside has a naphthalene moiety (EDANS) as a fluorescent energy donor at the reducing end and a 4-(4'-dimethylaminobenzeneazo)-benzene derivative as an acceptor chromophore at the non-reducing end. The key steps for the preparation of the target molecule involved transglycosylation reactions of cellobiosyl and cellotetraosyl fluoride donors onto cellobiosyl acceptors catalysed by the E197A mutant of cellulase Cel7B from Humicola insolens. Upon digestion with various cellulases, the energy transfer was disrupted and an increase of fluorescence was observed. PMID- 11921223 TI - Waste-free chemistry of diazonium salts and benign separation of coupling products in solid salt reactions. AB - Gas-solid and solid-solid techniques allow for waste-free and quantitative syntheses in the chemistry of diazonium salts. Five techniques for diazotations with the reactive gases NO(2), NO and NOCl are studied. Two types are mechanistically investigated with atomic force microscopy (AFM) and are interpreted on the basis of known crystal packings. The same principles apply to the cascade reactions that had been derived from one-step reactions. Solid diazonium salts couple quantitatively with solid diphenylamine and anilines to give the triazenes. Azo couplings are achieved with quantitative yields by cautious co-grinding of solid diazonium salts with beta-naphthol and C-H acidic heterocycles, such as barbituric acids or pyrazolinones. Solid diazonium salts may be more easily applied in a stoichiometric ratio for couplings in solution. Co-grinding of solid diazonium salts with KI gives quantitative yields of various solid aryl iodides. The unavoidable coupling products in salt reactions are completely separated from the insoluble products in a highly benign manner. The solid-state reactions compare favourably with similar solution reactions that produce much waste. The structures of the products are elucidated with IR and NMR spectroscopy and mass spectrometry, while the tautomeric properties of the compounds are studied with density functional calculations at the B3LYP/6-31G* and BLYP/6-31G** levels. PMID- 11921224 TI - Preparation and characterization of DNA films induced by UV irradiation. AB - Large amounts of DNA-enriched materials, such as salmon milts and shellfish gonads, are discarded as industrial waste. We have been able to convert the discarded DNA to a useful material by preparing novel DNA films by UV irradiation. When DNA films were irradiated with UV light, the molecular weight of DNA was greatly increased. The reaction was inhibited by addition of the radical scavenger galvinoxyl suggesting that the DNA polymerization with UV irradiation proceeded by a radical reaction. Although this UV-irradiated DNA film was water-insoluble and resistant to hydrolysis by nuclease, the structure of the DNA film in water was similar to non-irradiated DNA and maintained B-form structure. In addition, the UV-irradiated DNA film could effectively accumulate and condense harmful DNA-intercalating compounds, such as ethidium bromide and acridine orange, from diluted aqueous solutions. The binding constant and exclusion number of ethidium bromide for UV-irradiated DNA were determined to be 6.8 +/- 0.3 x 10(4) M(-1) and 1.6 +/- 0.2, respectively; these values are consisted with reported results for non-irradiated DNA. The UV-irradiated DNA films have potential uses as a biomaterial filter for the removal of harmful DNA intercalating compounds. PMID- 11921225 TI - Hybrid composites of monodisperse pi-conjugated rodlike organic compounds and semiconductor quantum particles. AB - Composite materials of quantum particles (Q-particles) arranged in layers within crystalline powders of pi-conjugated, rodlike dicarboxylic acids are reported. The synthesis of the composites, either as three-dimensional crystals or as thin films at the air-water interface, comprises a two-step process: 1) The preparation of the Cd salts 6 (Cd), 8 (Cd) or Pb salts 6 (Pb), 8 (Pb) of the oligo(p-phenyleneethynylene)dicarboxylic acids 6 (H), 8 (H), in which the metal ions are arranged in ribbons and are separated by the long axis of the organic molecules, as demonstrated by X-ray powder diffraction analysis of the solids and grazing incidence X-ray diffraction analysis of the films on water. 2) Topotactic solid/gas reaction of these salts with H(2)S to convert the metal ions into Q particles of CdS or PbS embedded in the organic matrix that consists of the acids 6 (H) and 8 (H). These hybrid materials have been characterized by X-ray photoelectron spectroscopy and transmission electron microscopy. PMID- 11921226 TI - Synthesis of a giant 222 carbon graphite sheet. AB - In this paper we present the synthesis and characterization of the so far largest polycyclic aromatic hydrocarbon (PAH), containing 222 carbon atoms or 37 separate benzene units. First a suitable three-dimensional oligophenylene precursor molecule is built up by a sequence of Diels-Alder and cyclotrimerization reactions and then planarized in the final step by oxidative cyclodehydrogenation to the corresponding hexagonal PAH. Structural proof is based on isotopically resolved MALDI-TOF mass spectra and electronic characteristics are studied by UV/Vis spectroscopy. PMID- 11921227 TI - Enantioselective hydrogenation over cinchona-modified Pt: the special role of carboxylic acids. AB - The influence of acetic acid (AcOH) and trifluoroacetic acid (TFA) on the hydrogenation of ethyl-4,4,4-trifluoroacetoacetate has been investigated by using Pt/Al(2)O(3) modified by cinchonidine and O-methylcinchonidine. We have shown that the sometimes dramatic changes in enantioselectivity and rate cannot simply be interpreted by protonation of the alkaloid modifier. We propose a new three step reaction pathway, involving interaction of the carboxylic acid with the reactant and the chiral modifier. The mechanism is supported by IR spectroscopic identification of cyclic TFA-modifier ion pairs. This new approach can rationalise the poorly understood role of acids in the enantioselective hydrogenation of activated ketones over cinchona-modified platinum metals. PMID- 11921228 TI - Surface organometallic chemistry of main group elements: selective synthesis of silica supported [triple bond Si-OB(C(6)F(5))(3)](-)[HNEt(2)Ph](+). AB - The reaction of the Lewis acid B(C(6)F(5))(3) with silanol groups of silica surfaces, dehydroxylated at different temperatures (300, 500, 700, and 800 degrees C), has been investigated in presence of the Bronsted base NEt(2)Ph. The structure of the resulting modified silica supports [triple bond Si OB(C(6)F(5))(3)](-)[HNEt(2)Ph](+) (1) has been carefully identified by IR and multinuclear solid-state NMR spectroscopies, isotopic (2)H and (18)O labeling, elemental analysis, molecular modeling, and comparison with synthesized molecular models. Highly dehydroxylated silica surfaces were required to transform selectively each silanol group into unique [triple bond Si-OB(C(6)F(5))(3)]( )[HNEt(2)Ph](+) fragments. For lower dehydroxylation temperatures, two sorts of surface sites were coexisting on silica: the free silanol groups [triple bond SiOH] and the ionic species 1. PMID- 11921229 TI - Synthesis of functionalized 2-alkylidenetetrahydrofurans by cyclization of 1,3 bis(trimethylsilyloxy)-1,3-butadienes with epoxides. AB - The Lewis acid mediated cyclization of epoxides with 1,3-bis(trimethylsilyloxy) 1,3-butadienes, electroneutral equivalents of 1,3-dicarbonyl dianions, results in the formation of 2-alkylidenetetrahydrofurans with a great variety of substitution patterns and functional groups. This includes the synthesis of 2,3' bifuranylidenes and 7-oxabicyclo[4.3.0]nonanes. The cyclization of dienes with functionalized epoxides containing base-labile groups proceeds with good chemoselectivity. In all reactions, good regio- and E diastereoselectivities are observed. Based on the stereoselectivities observed for reactions of 1,2 disubstituted epoxides, a working hypothesis for the mechanism of the reaction is suggested. PMID- 11921230 TI - Chemically induced contraction and stretching of a linear rotaxane dimer. AB - Copper(I)-induced assembly of two self-complementary identical units, which consist of a ring that incorporates a 1,10-phenanthroline group attached to a small filament containing a second 1,10-phenanthroline (phen) group, leads quantitatively to a doubly threaded complex. Each copper(I) center is four coordinate and is located inside a ring and bound to a phen from the macrocyle. The two other coordination sites are occupied by a phen from the filament connected to the other ring. An X-ray structure of the dicopper(I) complex unambiguously demonstrates the doubly threaded nature of the system. The molecule has C(2) symmetry in the crystal. This is an extended form with a Cu small middle dot small middle dot small middle dotCu separation of 18.3 A and an overall length close to 40 A. Further synthetic work, which utilizes the two terminal phenolic functions of the previous dicopper(I) complex, gives rise to a more complex system in which both filaments have been prolonged in opposite directions by 2,2':6',2"-terpyridine (terpy) motifs and bulky stoppers. The organic backbone is that of a rotaxane dimer. Although redox cycling of Cu(I) to Cu(II) did not lead to intramolecular rearrangement, simple chemical reactions induced large conformational changes. The rotaxane dimer was set in motion as follows. The dicopper(I) complex, which is in an extended conformation, was demetallated by using KCN. From the free ligand, the dizinc complex was formed quantitatively at room temperature. (1)H NMR data show that a new conformation is obtained: each Zn(II) is five-coordinate (phen + terpy), and the molecule is in a contracted conformation. This process is reminiscent of biological muscles in the sense that the two filaments of this system can be moved along one another in a gliding motion that keeps the whole system together, but which converts a stretched compound (overall length approximately equal to 83 A) into a contracted species (overall length approximately equal to 65 A, according to CPK models). The motion is quantitatively reversed by the addition of an excess of copper(I) to the dizinc complex; this regenerates the extended starting form. Although the motivation of the present contribution was to illustrate that a muscle-like molecule may be stretched or contracted using electrochemistry and coordination chemistry, the main body of the work is organic synthesis. This is testified by the fact that the dicopper(I) rotaxane dimer was obtained in 23 steps from commercially available compounds. PMID- 11921231 TI - Structural investigation of the HIV-1 envelope glycoprotein gp160 cleavage site. AB - The selective proteolytic activation of the HIV-1 envelope glycoprotein gp160 by furin and other precursor convertases (PCs) occurs at the carboxyl side of the sequence Arg508-Glu-Lys-Arg511 (site 1), in spite of the presence of another consensus sequence: Lys500-Ala-Lys-Arg503 (site 2). We report on the solution structural analysis of a 19-residue synthetic peptide, p498, which spans the two gp160-processing sites 1 and 2, and is properly digested by furin at site 1. A molecular model is obtained for p498, by means of molecular dynamics simulations, from NMR data collected in trifluoroethanol/water. The peptide N-terminal side presents a 9-residue helical segment, enclosing the processing site 2; the C terminal segment can be described as a loop exposing the processing site 1. A hypothesis for the docking of p498 onto the catalytic domain of human furin, modeled by homology and fitting previous site-directed mutagenesis studies, is also presented. p498 site 1 is shown to have easy access to the furin catalytic site, unlike the nonphysiological site 2. Finally, on the basis of available data, we suggest a possible structural motif required for the gp160-PCs recognition. PMID- 11921232 TI - Synthesis and characterization of phthalocyanines with direct Si-Si linkages. AB - Several cofacial phthalocyanines (Pcs) with an Si-Si linkage were obtained by one step condensation of 1H-isoindole-1,3(2H)-diimine with hexachlorodisilane as template in quinoline. They were characterized by gel-permeation chromatography, IR, NMR spectroscopy, and mass spectrometry, and cyclic voltammetry. The results strongly suggest that we indeed obtained Pc dimers directly linked by an Si-Si bond using this novel concept of utilizing a compound/salt with an element element bond as a template. The cofacial dimer structures are reasonably supported by X-ray absorption near-edge structure (XANES), electronic absorption and magnetic circular dichroism (MCD) spectra, and molecular orbital (MO) calculations. Interestingly, they show an electronic absorption spectrum very similar to that of silicon tetrabenztriazacorrole (SiTBC). PMID- 11921235 TI - Asymmetric halogeno-bridged complexes: new reagents in organometallic synthesis and catalysis. AB - Several methods to synthesize bimetallic complexes in which two different metal fragments are connected by halide bridges are described. Using simple starting materials a large pool of structurally defined bimetallic complexes with unique chemical reactivities can be prepared in short time. Applications in organometallic synthesis and homogeneous catalysis are discussed. PMID- 11921233 TI - Towards synthetic adrenaline receptors--shape-selective adrenaline recognition in water. AB - In spite of their key role in signal transduction, the mechanism of action of adrenergic receptors is still poorly understood. We have imitated the postulated binding pattern of the large membrane protein with a small, rationally designed synthetic host molecule. Experimental evidence is presented for the simultaneous operation of electrostatic attraction, hydrogen bonds, pi stacking, and hydrophobic interactions. By virtue of this combination of weak attractive forces, adrenaline derivatives in water are bound with high shape selectivity for the slim dopamine skeleton. We think that these findings support the postulated cooperative interplay of noncovalent interactions in the natural receptors. In addition, they provide access to a new type of adrenaline sensor. This may be the first step towards an artificial signal-transduction system. PMID- 11921236 TI - Three novel interpenetrating diamondoid networks from self-assembly of 1,12 dodecanedinitrile with silver(I) salts. AB - The self-assembly of 1,12-dodecanedinitrile (ddn) with various silver salts (NO(3)(-), PF(6)(-), AsF(6)(-), ClO(4)(-)) afforded new polymeric coordination networks with the general formula [Ag(ddn)(2)]X. All these species contain interpenetrating diamondoid nets showing interesting features: with X=NO(3)(-) the cationic [Ag(ddn)(2)](+) network exhibits the highest interpenetration (tenfold) ever found within diamondoid nets exclusively based on coordinative bonds. When X=PF(6)(-) or AsF(6)(-) an eightfold diamondoid network is obtained that shows an unusual [4+4] mode of interpenetration, instead of the "normal" set of eight nets equally translated along a principal axis of the adamantanoid cages. The polymeric species that forms with X=ClO(4)(-) is a fourfold interpenetrating diamondoid network; the lower degree of interpenetration in this case is related to the conformation assumed by the flexible ddn ligands. PMID- 11921237 TI - Synthesis and structural properties of patellamide A derivatives and their copper(II) compounds. AB - The synthesis, characterization and copper(II) coordination chemistry of three new cyclic peptide ligands, PatJ(1) (cyclo-(Ile-Thr-(Gly)Thz-Ile-Thr-(Gly)Thz)), PatJ(2) (cyclo-(Ile-Thr-(Gly)Thz-(D)-Ile-Thr-(Gly)Thz)), and PatL (cyclo-(Ile-Ser (Gly)Thz-Ile-Ser-(Gly)Thz)) are reported. All of these cyclic peptides and PatN (cyclo-(Ile-Ser-(Gly)Thz-Ile-Thr-(Gly)Thz)) are derivatives of patellamide A and have a [24]azacrown-8 macrocyclic structure. All four synthetic cyclic peptides have two thiazole rings but, in contrast to patellamide A, no oxazoline rings. The molecular structure of PatJ(1), determined by X-ray crystallography, has a saddle conformation with two close-to-coparallel thiazole rings, very similar to the geometry of patellamide D. The two coordination sites of PatJ(1) with thiazole-N and amide-N donors are each well preorganized for transition metal ion binding. The coordination of copper(II) was monitored by UV/Vis spectroscopy, and this reveals various (meta)stable mono- and dinuclear copper(II) complexes whose stoichiometry was confirmed by mass spectra. Two types of dinuclear copper(II) complexes, [Cu(2)(H(4)L)(OH(2))(n)](2+) (n=6, 8) and [Cu(2)(H(2)L)(OH(2))(n)] (n=4, 6; L=PatN, PatL, PatJ(1), PatJ(2)) have been identified and analyzed structurally by EPR spectroscopy and a combination of spectra simulations and molecular mechanics calculations (MM-EPR). The four structures are similar to each other and have a saddle conformation, that is, derived from the crystal structure of PatJ(1) by a twist of the two thiozole rings. The small but significant structural differences are characterized by the EPR simulations. PMID- 11921238 TI - Reactivity of 1,6-bis(trimethylsilyl)-hexa-3-ene-1,5-diynes towards triethylborane, triallylborane, and 1-boraadamantane: first molecular structure of a 4-methylene-3-borahomoadamantane derivative, and the first 6,8 diborabicyclo[2.2.2]oct-2-ene derivative. AB - Compounds (E)- (1) and (Z)-1,6-bis(trimethylsilyl)-hexa-3-ene-1,5-diyne (2) react with triethylborane (3) by 1,1-ethylboration in a 1:1 or 1:2 molar ratio (in the case of 1), whereas in the case of 2 only the 1:1 product is formed. The analogous reactions of 1 or 2 with triallylborane (4) are more complex because of competition between 1,1-allyl- and 1,2-allylboration. Again, compound 2 reacts only with one equivalent of 4. In the case of 1-boraadamantane (5), 1,1 organoboration of 1 and 2 takes place either at one or at both C[triple bond]C bonds leading to compounds containing the 4-methylene-3-borahomoadamantane unit(s). The product of the reaction of 1 with two equivalents of 5 was characterized by X-ray structure analysis. The primary products of the reaction of 2 with 5 rearrange upon heating by deorganoboration and organoboration to give finally a tetracyclic compound 24 that contains an exocyclic allenylidene group. The product of the 1:2 reaction of 2 with 5 rearranges to give the 6,8-dibora bicyclo[2.2.2]oct-2-ene derivative 25. All reactions were monitored by (1)H, (11)B, (13)C, and (29)Si NMR spectroscopy. PMID- 11921239 TI - A helical peptide receptor for [60]fullerene. AB - Two terminally blocked nonapeptides, each made up of six Aib residues, a Gly spacer and two L-Tyr residues in positions 2 and 8 (these are substituted in the side chain with either ferrocenoyl or methyl moieties), have been synthesized by solution methods and fully characterized. FT-IR absorption and two-dimensional NMR analyses indicate that a 3(10)-helical conformation is adopted by these rigid peptides in structure-supporting solvents. An X-ray diffraction investigation shows that the bis-L-Tyr(Me) nonapeptide in the crystal state is folded in a regular right-handed 3(10)-helical structure. As five amino acid units separate the two substituted L-Tyr residues in the peptide sequence, the two side chain moieties will-in solution-face each other after two complete turns of the ternary helix. By carrying out a detailed photophysical analysis, we have demonstrated that the electron-rich, hydrophobic and wide cavity generated by the nonapeptide template with two ferrocenoyloxybenzyl walls is able to host [60]fullerene. Further evidence for this superstructure has been provided in the gas phase by a mass spectrometric investigation. PMID- 11921240 TI - Different quantitation approaches for xenobiotics in human urine samples by liquid chromatography/electrospray tandem mass spectrometry. AB - The potential of liquid chromatography combined with tandem mass spectrometry (LC/MS/MS) for the determination of pesticide metabolites in human urine at the sub-ppb level is explored. Metabolites from two organophosphorous pesticides, 4 nitrophenol (from parathion and parathion-methyl) and 3-methyl-4-nitrophenol (from fenitrothion), are taken as model analytes to conduct this study. After direct injection of the urine sample (10 microL), different approaches were evaluated in order to achieve correct quantitation of analytes using an electrospray ionisation (ESI) interface. Thus, the feasibility of using external calibration was checked versus the use of different isotope-labeled internal standards. The advantages of applying coupled-column liquid chromatography (LC/LC) as an efficient clean-up without any type of sample manipulation are also discussed. The combination of LC/LC with ESI-MS/MS allows the direct analysis of free metabolites in urine, as the automated clean-up performed by the coupled column technique is sufficient for the removal of interferences that suppress the ionisation of analytes in the ESI source. Using this procedure with external calibration, good precision and recoveries, and detection limits below 1 ng/mL are reached with analysis run times of around 8 min. The hyphenated technique LC/LC/ESI-MS/MS is proved to be a powerful analytical tool, allowing the rapid, sensitive and selective determination of 4-nitrophenol and 3-methyl-4-nitrophenol in human urine without any sample treatment. PMID- 11921241 TI - Gas chromatography/negative-ion chemical ionisation mass spectrometry for the quantitative analysis of morphine in human plasma using pentafluorobenzyl carbonate derivatives. AB - A sensitive and specific method for the quantitative determination of morphine in human plasma is presented. Morphine was extracted from plasma by solid phase extraction on C18 and converted to its pentafluorobenzyl carbonate trimethylsilyl derivative. The derivatives were analysed without further purification. Using gas chromatography/negative ion chemical ionisation mass spectrometry, a useful diagnostic fragment ion at m/z 356 is obtained at high relative abundance. Deuterated morphine was used as internal standard. Calibration graphs were linear within the range 1.25 to 320 nmol/L. Intra-day precision was 3.82% (15 nmol/L), 2.85% (75 nmol/L) and 4.13% (225 nmol/L), inter-day variability was found to be 1.77% (15 nmol/L), 4.95% (75 nmol/L) and 9.88% (225 nmol/L). Inter-day accuracy showed deviations of 2.18% (15 nmol/L), -0.72% (75 nmol/L) and -0.13% (225 nmol/L). The method is rugged and robust and has been applied to the batch analysis of morphine during pharmacokinetic profiling of the drug. PMID- 11921242 TI - Quantitative determination of perfluorooctanoic acid ammonium salt in human serum by high-performance liquid chromatography with atmospheric pressure chemical ionization tandem mass spectrometry. AB - A sensitive, specific, accurate and reproducible analytical method was developed and validated to quantify perfluorooctanoic acid (PFOA) in human serum. After initial extraction with an ion-paring reagent, the procedure for quantifying PFOA is based on high-performance liquid chromatography (HPLC) interfaced to negative ion tandem mass spectrometry, operating in selected ion monitoring mode. The retention times of PFOA and its internal standard (D,L-malic acid) were 5.85 and 1.70 min, respectively. The assay was linear over the range 0-500 ng/mL, with a lower limit of quantification (LOQ) of 25 ng/mL, and with a coefficient of variation (CV) of 7.3%. The lower limit of detection (LOD) was assessed as 10 ng/mL. The overall precision and accuracy were assessed on three different days. The within- and between-day precision was < or =9.7 and 6.8%, respectively, and the accuracy was in the range 96-114%. The mean extracted recovery assessed at three different concentrations (100, 250, and 500 ng/mL) was always more than 85%. With this method no derivatization procedure was needed, thus avoiding possible thermal and chemical decomposition reactions of PFOA. The assay was applied to quantify perfluorooctanoic acid in serum from employees exposed to fluorochemicals commonly used in industrial applications for polymer production. The quantitative results for PFOA blood levels were found to vary between 100 and 982 ng/mL. PMID- 11921243 TI - Determination of phenolic compounds in rose hip (Rosa canina) using liquid chromatography coupled to electrospray ionisation tandem mass spectrometry and diode-array detection. AB - Liquid chromatography coupled with negative and positive electrospray ionisation (ESI) tandem mass spectrometry (MS/MS) and diode-array detection (DAD) was used for determination of phenols in rose hip (Rosa canina) extract. ESI mass spectra of the chromatographically separated phenols gave the molecular weight of the compounds through prominent [M - H](-) ions for most of the compounds and M(+) ions for the anthocyanins. Collision induced dissociation (CID) of the [M - H](-) (or M(+)) precursor ions yielded product ions which determined the molecular weight of the aglycones. In-source fragmentation followed by CID of the resulting deprotonated aglycone ([A - H](-)) provided product ions for the identification of the unconjugated phenols. The identification was based on comparison with product ion spectra of commercial standards. UV-diode-array spectra were used for identity confirmation. This combined approach allowed the identification in rose hip extract of an anthocyanin, i.e. cyanidin-3-O-glucoside, several glycosides of quercetin and glycosides of taxifolin and eriodictyol. Phloridzin was identified, and several conjugates of methyl gallate were also found, one of which was tentatively identified as methyl gallate-rutinoside. Catechin and quercetin were found as the aglycones in the extract. PMID- 11921244 TI - Determination of selected sulfonamide antibiotics and trimethoprim in manure by electrospray and atmospheric pressure chemical ionization tandem mass spectrometry. AB - A method for the determination of sulfonamides and trimethoprim in the complex matrix liquid manure has been developed using reversed-phase liquid chromatography and atmospheric pressure chemical ionization (APCI) tandem mass spectrometry. A comparison was made between electrospray and atmospheric pressure chemical ionization. APCI proved to be more robust and less sensitive to matrix effects. High-performance liquid chromatographic (HPLC) separation of the analytes was achieved in less than 7 min. The compounds were extracted with ethyl acetate and the extracts were cleaned up by solid-phase extraction on an aminopropyl column. Recoveries were not dependent on the concentration level. The mean recoveries were as follows: trimethoprim 79.0%, sulfadiazine 80.5%, N(4) acetylsulfadiazine 91.0%, sulfamerazine 78.6%, sulfadimidine 77.2% and sulfamethoxazole 82.8%. Linearity was established over a concentration range of 5 to 5000 microg/kg with correlation coefficients greater than 0.99. The method had a limit of quantitation (LOQ) of 5 microg/kg manure. PMID- 11921245 TI - Two-dimensional separations with electrospray ionization ambient pressure high resolution ion mobility spectrometry/quadrupole mass spectrometry. AB - The coupling of ion mobility spectrometry (IMS) instruments with mass spectrometers has been described since early in IMS development, most commonly with quadrupole mass analyzers. The recent development of IMS with time-of-flight (TOF) instruments has demonstrated that the time compatibility (IMS milliseconds and TOFMS microseconds) of the two techniques enables rapid two-dimensional separations to be performed, theoretically in the order of seconds for a complete analysis. This study presents a unique way to operate a traditional IMS/QMS system to attain separations similar to those achieved with IMS/TOF. For this new approach, the quadrupole was slowly scanned in the single-ion monitoring mode while IMS spectra were continually embedded in each m/z step. In this way, two dimensional separations (IMS drift times and m/z) were obtained using the traditional IMS/QMS arrangement. An example of a five amino acid separation (quadrupole scan of 40 m/z values at a rate of approximately 7 steps/min) led to a complete two-dimensional analysis within 6 min, comparable to rapid chromatographic separations with mass spectrometry. Proposed approaches to reduce the analysis time are discussed and a reduction in the analysis time to less than 1 min is feasible when the IMS/QMS separation conditions are optimized. PMID- 11921246 TI - Tandem mass spectra of tryptic peptides at signal-to-background ratios approaching unity using electrospray ionization high-field asymmetric waveform ion mobility spectrometry/hybrid quadrupole time-of-flight mass spectrometry. AB - High-field asymmetric waveform ion mobility spectrometry (FAIMS) has been coupled to a quadrupole time-of-flight mass spectrometer for the tandem mass spectrometric analysis of tryptic peptides of pig hemoglobin. Using FAIMS, low levels (fmol/microL) of multiply charged tryptic peptides were separated from relatively intense chemical background such that their tandem mass spectra (MS/MS) lacked many background-related fragment ions observed using a conventional ESI-QqTOFMS instrument. Substantial improvements in both first-order and tandem mass spectra were realized while maintaining approximately the same absolute intensities. PMID- 11921247 TI - Atmospheric pressure laser desorption/chemical ionization mass spectrometry: a new ionization method based on existing themes. AB - We have designed and constructed an atmospheric pressure laser desorption/chemical ionization (AP-LD/CI) source that utilizes a laser pulse to desorb intact neutral molecules, followed by chemical ionization via reagent ions produced by a corona discharge. This source employs a heated capillary atmospheric pressure inlet coupled to a quadrupole ion trap mass spectrometer and allows sampling under normal ambient air conditions. Preliminary results demonstrate that this technique provides approximately 150-fold increase in analyte ions compared to the ion population generated by atmospheric pressure infrared matrix-assisted laser desorption/ionization (AP-IR-MALDI). PMID- 11921248 TI - Gas-phase ion/molecule reactions between dimethoxyphosphonium ions and aromatic hydrocarbons. AB - Ion/molecule reactions between O=P(OCH(3))(2)(+) phosphonium ions and six aromatic hydrocarbons (benzene, toluene, 1,2,4-trimethylbenzene, naphthalene, acenaphthylene and fluorene) were performed in a quadrupole ion trap mass spectrometer. The O=P(OCH(3))(2)(+) phosphonium ions, formed by electron impact from neutral trimethyl phosphite, were found to react with aromatic hydrocarbons (ArHs) to give (i) an adduct [ArH, O=P(OCH(3))(2)](+) and (ii) for ArHs which have an ionization energy below or equal to 8.14 eV, a radical cation ArH(+ *) by charge transfer reaction. Collision-induced dissociation experiments, which produce fragment ions other than the O=P(OCH(3))(2)(+) ions, indicate that the adduct ions are covalent species. Isotope-labeled ArHs were used to elucidate fragmentation mechanisms. The charge transfer reactions were investigated using density functional theory at the B3LYP/6-311 + G(3df,2p)//B3LYP/6-31G(d,p) level of theory. The potential energy surface obtained from B3LYP/6-31G(d,p) calculations for the reaction between O=P(OCH(3))(2)(+) and benzene is described. PMID- 11921249 TI - Post-source decay time-of-flight study of fragmentation mechanisms of protonated synthetic polymers under matrix-assisted laser desorption/ionization conditions. AB - Post-source decay (PSD) of three different nylon oligomers desorbed under matrix assisted laser desorption/ionization (MALDI) conditions was studied and their fragmentation pathways were investigated. The fragmentation of the protonated oligomers is very similar to that of peptides. The b(n)(+), y(n)(+) and z(n)(+) series of ions were observed in abundance in the PSD spectrum. The end groups and the length of the spacer in the repeating unit influence the fragmentation of the different polyamides and the relative abundances of the product ions. Competitive dehydration and deamination reactions were observed, and depend on the nature of the end groups and the repeating units. The PSD spectra are very similar to collision-induced dissociation (CID) spectra obtained under low-energy conditions, implying that the selected precursor ions possess similar average internal energies. All the peaks observed in the PSD spectrum can be rationalized by reasonable fragmentation mechanisms. PMID- 11921250 TI - Liquid chromatography/electrospray ionisation mass spectrometric investigations of imidazoline corrosion inhibitors in crude oils. AB - The so-called imidazolines (2-alkyl-1-[ethylalkylamide]-2-imidazolines and 2 alkyl-1-ethylamine-2-imidazolines) are a group of surface-active compounds, complex mixtures of which are used by various industries as surfactants and corrosion inhibitors. Although their industrial synthesis was reported over 100 years ago, few methods for the determination of individual imidazolines in mixtures, including industrial matrices such as crude oils, have been reported. Here we demonstrate that spiking of crude oils with synthetic imidazolines followed by solid-phase extraction and liquid chromatography/electrospray ionisation multistage mass spectrometry (LC/ESI - MS(n)) allows an estimation of low (<10) parts per million concentrations of individual imidazolines in crude oils. Whilst non-optimised at present, the method is a significant advance and may prove useful not only for improving an understanding of the mechanisms of industrial imidazoline synthesis and for monitoring downhole and topside oilfield operations, but also for the determination of the fate of imidazoline-based oilfield corrosion inhibitors and surfactants in the environment. PMID- 11921251 TI - Backbone cleavages of [M - H](-) anions of peptides. Cyclisation of citropin 1 peptides involving reactions between the C-terminal [CONH](-) residue and backbone amide carbonyl groups. A new type of beta cleavage: a joint experimental and theoretical study. AB - This paper reports the study of backbone cleavages in the collision-induced negative-ion mass spectra of the [M - H](-) anions of some synthetic modifications of the bioactive amphibian peptide citropin 1 (GLFDVIKKVASVIGGL NH(2)). The peptides chosen for study contain no amino acid residues which could effect facile side-chain cleavage, i.e. Ser (-CH(2)O, side-chain cleavage) and Asp (-H(2)O) are replaced by Ala or Lys. We expected that such peptides should exhibit standard and pronounced peaks due to alpha cleavage ions (and to a lesser extent beta cleavage ions) in their collision-induced negative-ion spectra. This expectation was realised, but the spectra also contained peaks formed by a new series of cleavage anions. These are produced following cyclisation of the C terminal CONH(-) moiety at carbonyl functions of amide groups along the peptide backbone; effectively transferring the NH of the C-terminal CONH(-) group to other amino acid residues. We have called the product anions of these processes beta' ions, in order to distinguish them from standard beta ions. Some beta' ions also fragment directly to some other beta' ions of smaller mass. The reaction coordinates of alpha,beta and beta' backbone processes have been calculated at the HF/6-31G*//AM1 level theory for simple model systems. The initial cyclisation step of the beta' sequence is barrierless and exothermic. Subsequent steps have a maximum barrier of +40 kcal mol(-1), with the overall reaction being endothermic by some 30 kcal mol(-1) at the level of theory used. These calculations take no account of the complexity of the conformationally flexible peptide system, and it is surprising that each of the two reacting centres can 'find' each other in such a large system. PMID- 11921252 TI - Electrospray mass spectrometric studies of L-carnosine (beta-alanyl-L-histidine) complexes with copper(II) or zinc ions in aqueous solution. AB - Electrospray ionization mass spectrometry (ESI-MS) was used for the speciation of supramolecular assemblies formed between equimolar amounts of carnosine and copper or zinc ions in dilute aqueous solutions. In the case of pure carnosine and carnosine/copper systems, the effect of pH changes, in the range 2-9, on the complexes surviving in solution was also explored. ESI data, besides supporting previous reported results on the formation of dimeric carnosine/copper and carnosine/zinc complexes, allowed a more complete speciation of the examined systems, bringing to light the existence of bis-complex species and, in the zinc case, the formation of oligomeric species. The data obtained for the systems investigated show that ESI-MS is not only a reliable and fast technique for the analysis of the metal/ligand systems, but also an interesting tool to obtain stoichiometric information on metal complexes formed in very low concentration solutions. PMID- 11921253 TI - Detection of solid cobalt species by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry. PMID- 11921254 TI - Study of polycondensation reactions of Ge(OEt)(4) and Ge(OEt)(4)/Si(OEt)(4) by electrospray ionization mass spectrometry. PMID- 11921255 TI - Use of carbon dioxide and ammonia as nebuliser gases in electrospray ionisation mass spectrometry. PMID- 11921258 TI - Electrospray ionisation and ion-trap fragmentation of substituted 3,4-dihydro 2(1H)-pyridin-2-ones. AB - A variety of 5-alkoxycarbonyl-4-aryl-6-methyl-3,4-dihydro-2(1H)-pyridones and hexahydrofuro[3,4-b]-2(1H)-pyridones have been investigated by electron impact (EI) and electrospray ionisation (ESI) techniques. Sequential product ion fragmentation (MS(n)) was performed to elucidate the degradation pathways for these compounds. Comparisons are made between positive and negative even-electron ions from ESI spectra and the molecular radical cations obtained under EI conditions. The data collected in this paper provide information on the strong impact that different substituents have on the ion fragmentation process. PMID- 11921257 TI - Analysis of saccharides in beer samples by flow injection with electrospray mass spectrometry. AB - Saccharides in foods play important roles, as they are essential substrates for fermentation processes. In brewing, the concentration of maltooligosaccharides influences the characteristics of beers and therefore their determination is of great practical interest. Electrospray ionization mass spectrometry (ESI-MS) was applied to identify and characterise maltooligosaccharides in beer samples. The effects due to different cation concentrations and dilution of samples were studied. Furthermore, quantitative analyses of maltooligosaccharides by means of flow-injection ESI-MS (FI/ESI-MS) of 1-microL beer samples (diluted 1000-fold) are described. PMID- 11921259 TI - Double resonance ejection in a micro ion trap mass spectrometer. AB - Ion ejection from a cylindrical micro ion trap by resonance excitation of the secular motion is observed to be strongly dependent on the frequency of the secular motion at resonance. Both the intensity of the ion signal and the mass resolution of the resulting mass spectrum are increased when the ion secular frequency is approximately that of a nonlinear resonance of the trap. The resonances are attributed to electrical as well as geometrical considerations. PMID- 11921260 TI - Mass spectrometric analysis of cyclofenil and its metabolites in human urine. AB - Using gas chromatography/electron impact-mass spectrometry (GC/EI-MS) and high performance liquid chromatography with atmospheric pressure chemical ionization tandem mass spectrometry (HPLC/APCI-MS/MS), the structures of cyclofenil metabolites in human urine have been assigned. The hydroxyl metabolites liberated from the glucuronide conjugates after acid hydrolysis were characterized as the trimethylsilyl (O-TMS) derivatives using GC/MS. The conjugate glucuronide forms were detected without hydrolysis by HPLC/MS. Cyclofenil was not observed in urine. Tentative structures for the two metabolites are proposed. PMID- 11921261 TI - Determination of species-specific components in the venom of Parabuthus scorpions from southern Africa using matrix-assisted laser desorption time-of-flight mass spectrometry. AB - The aim of the present study was to analyze mass spectra of scorpions belonging to the genus Parabuthus (Pocock 1890) by means of matrix-assisted laser desorption time-of-flight mass spectrometry (MALDI-TOFMS) and to construct a species-specific venom code for species identification. The venom compositions of sixteen Parabuthus species, occurring in southern Africa, were characterized using representative peaks in the molecular mass range of 6400-8400 Da. This mass range is characteristic for the typical long-chain neurotoxins influencing sodium channels. Only a few of these peptides have been sequenced up to now. The impetus for development of these species-specific profiles was the observation of unique, highly reproducible mass spectral peaks within a specific species. An identification label for all the different species could be found using a minimum number of peaks. MALDI-TOFMS is therefore proposed as a complementary method to morphological and behavioural characteristics for species and ultimately subspecies discrimination. PMID- 11921262 TI - Pyrolysis-gas chromatography/mass spectrometry of fractions separated from a low temperature coal tar: an attempt to develop a general method for characterising structures and compositions of heavy hydrocarbon liquids. AB - A low-temperature coal tar has been fractionated by column chromatography into acetonitrile, pyridine and 1-methyl-2-pyrrolidinone- (NMP) solubles. The tar and its fractions have been examined by pyrolysis-gas chromatography/mass spectrometry (GC/MS). Fractionation by planar chromatography was also carried out for purposes of comparison. Molecular masses of the fractions were estimated by size-exclusion chromatography (SEC), and bulk structural characterisation was carried out by (13)C-NMR and UV-fluorescence spectrometry. SEC showed that the fractions shifted to progressively shorter elution times (higher apparent masses) with diminishing solubility, i.e. from acetonitrile to NMP solubles. UV fluorescence spectra showed parallel shifts to longer wavelengths and lower fluorescence quantum yields, indicating increasing sizes of aromatic ring systems and increasingly complex molecules. GC/MS analysis of the tar showed alkanes from C10 to C32 and extensive series of alkylated aromatics, phenols, indenes, naphthalenes, phenanthrenes and fluoranthenes. Pyrolysis-GC/MS results for the acetonitrile solubles closely resembled the data for the tar sample, with extensive series of alkylated benzenes, phenols and naphthalenes as well as alkanes from C16 to C28. The pyridine-soluble fraction showed no significant aromatic pyrolysis products and only relatively weak signals for alkanes between C16 and C27. The NMP-soluble fraction showed even less overall signal, with no significant aromatic components and weak signals for alkanes between C21 and C25, even though (13)C-NMR analyses showed that approximately half of the carbon detected was aromatic. The aliphatics are assumed to provide bridging structures between polycyclic aromatic (PCA) ring systems. PMID- 11921263 TI - Self-esterification of fulvic acid model compounds in methanolic solvents as observed by electrospray ionization mass spectrometry. AB - The self-esterification of two fulvic acid model compounds in methanolic solvents was studied by electrospray ionization mass spectrometry (ESI-MS). The strongly acidic tetrahydrofurantetracarboxylic acid rapidly self-esterified to form mono- and dimethyl esters when stored in methanol, even at reduced temperatures. The weakly acidic analogue, cyclopentanetetracarboxylic acid, reacted minimally under the same conditions. The use of 50:50 methanol/water as a solvent reduced self esterification of the strong acid. However, the presence of water promoted the formation of multiply charged ions in the ESI mass spectra. The use of water and 50:50 acetonitrile/water as solvents eliminated self-esterification but the mass spectra still contained multiply charged ions. This study implies that the use of methanolic solvents with humic substances may compromise analytical data through the formation of methyl esters. PMID- 11921264 TI - Negative-ion electrospray ionization tandem mass spectrometry of N-phosphoryl amino acids and dipeptides. AB - The negative-ions of N-phosphoryl amino acids were studied by electrospray ionization tandem mass spectrometry (ESI-MS/MS). The negative-ion ESI-MS/MS of N phosphoryl amino acids showed characteristic fragmentation patterns different from those observed in the corresponding positive-ion ESI-MS/MS and negative-ion fast-atom bombardment mass spectra. For negative-ion ESI-MS/MS, a unique fragmentation from the N-terminal of N-phosphoryl amino acids or peptides containing a free beta-OH or CO(2)H group was observed to yield the characteristic fragment ion (RO)(2)P(O)O(-). The ease of the rearrangement depended on the position of the hydroxyl group in amino acids or peptides, and the N --> O rearrangement mechanism was proposed to involve the participation of the hydroxyl group. From previous solution-phase experiments and theoretical calculations, it was found that the beta-OH group was more active than gamma-OH, and the corresponding difference in negative-ion ESI-MS/MS was consistent with those previous findings. PMID- 11921265 TI - Stable isotope compositions of CO(2) in background air and at polluted sites in Hungary. AB - CO(2) samples were collected from air at three sites in Hungary for comparison of polluted and background areas. In order to reduce the uncertainties caused by the varying amount of N(2)O, a gas chromatography (GC)-based vacuum separation was applied. The reliability of the procedure was demonstrated by careful standardization and comparison with global network data. The stable isotope data show complex diurnal and seasonal variations that can be explained by fractionations during photosynthesis and respiration. The isotopic characteristics of pollution-derived (anthropogenic) and biogenic CO(2) appear to be indistinguishable at the study sites. However, the sites at unpolluted areas reveal a seasonal variation in the carbon isotope composition of biogenic CO(2) that may be related to changes in soil biogenic activities. The atmospheric background CO(2) shows constant delta(13)C in the region. Finally, the study demonstrates the need for careful standardization of sampling in order to make the data obtained from different sampling systems comparable. PMID- 11921266 TI - Analysis of a basic drug by on-line solid-phase extraction liquid chromatography/tandem mass spectrometry using a mixed mode sorbent. AB - Two on-line configurations using multiple 6- and 10-port valves were investigated for high-flow on-line extraction of a basic drug in rat plasma and human urine using a reversed-phase and a cation-exchange SPE sorbent. The first configuration studied was a single reversed-phase extraction cartridge (2.1 x 20 mm, 25 microm) using an optimized washing protocol. The results showed that up to 1.5 mL of sample (urine or plasma diluted 1:1) can be injected with limits of quantification (LOQs) as low as 100 pg/mL. The second configuration studied was the combination of a cation exchanger and a reversed-phase cartridge using at column dilution. The results showed better LOQs than those obtained with the single cartridge at 10 pg/mL with the same injection volume. The mass spectrometer was operated in the multiple reaction monitoring (MRM) mode. All calibration curves gave an average of 5% relative standard deviation (RSD). PMID- 11921267 TI - Hyphenation of capillary high-performance liquid chromatography with matrix assisted laser desorption/ionization time-of-flight mass spectrometry for nano scale screening of single-bead combinatorial libraries. AB - This paper focuses on the technical aspects of chemical screening from 384-well plate nano-scale single-bead combinatorial libraries. The analytical technique utilized is a combination of capillary liquid chromatography with ultraviolet detection and matrix-assisted laser desorption/ionization (MALDI) mass spectrometry. The HPLC/MALDI-MS hyphenation is achieved by means of a micro fraction collector with a peak detection system that automatically collects the peaks onto the MALDI targets for subsequent characterization. Several experimental parameters such as type of 384-well plate, well-plate sealing foils, and a column-switching procedure were investigated using a small test library of nine components. Additionally, the influence of different MALDI matrices, different MALDI targets and sample-spotting techniques on the MALDI detection sensitivity as well as the ruggedness and sample throughput capacity of this technique were studied. Optimum results for the analytes investigated were obtained with 2,5-dihydroxybenzoic acid using on-line mixing of HPLC effluent and matrix solution. To demonstrate the potential of this capillary HPLC/MALDI-TOFMS method, its application to several single-bead libraries was investigated. The instrumental method allowed for the rapid identification and purity assessment of combinatorial libraries with detection limits down to the higher femtomole level using both UV detection and MALDI mass spectrometry. PMID- 11921269 TI - International workshop on the relationship of prior therapy to balanced chromosome aberrations in therapy-related myelodysplastic syndromes and acute leukemia: overview report. PMID- 11921270 TI - Data management and statistical methods used in the analysis of balanced chromosome abnormalities in therapy-related myelodysplastic syndromes and therapy related acute leukemia: report from an international workshop. PMID- 11921271 TI - 11q23 balanced chromosome aberrations in treatment-related myelodysplastic syndromes and acute leukemia: report from an international workshop. AB - Among 511 patients with therapy-related myelodysplastic syndrome or acute leukemia (t-MDS/t-AL) and balanced chromosome aberrations, 162 (32%) had translocations involving 11q23. The recurring translocation partners were 9p22 (48%), 19p13.3 (11%), 19p13.1 (10%), 4q21 (9%), 6q27 (6%), 1p32 (2%), 16p13.1 (2%), 10p13 (1%), and 17q25 (1%); in 9%, the translocations were seen only once. The remaining 349 patients were divided into five subgroups based on the balanced aberration: 21q22, inv(16), t(15;17), Rare, and Unique aberrations. Patients in the 11q23 subgroup had a sole cytogenetic abnormality more often than those in the 21q22, inv(16), Rare, and Unique subgroups, and a complex karyotype or 5/del(5q) and/or -7/del(7q) less often than patients in the 21q22, Rare, and Unique subgroups. Clinically, 11q23 patients had acute lymphoblastic leukemia (ALL) more often as their primary disease and a shorter latency from start of treatment for the primary disease to their t-MDS/t-AL diagnosis, except when compared with the inv(16) subgroup. The 11q23 subgroup demonstrated a younger age at t-MDS/t-AL diagnosis, but this finding was not significant when patients with AL as their primary diagnosis were excluded. Survival from the time of diagnosis of t-MDS/t-AL was significantly shorter for the 11q23 subgroup compared with that of the 21q22, inv(16), and t(15;17) subgroups (median 8 vs. 14, 28, and 29 months, respectively). Inferior survival occurred even though 11q23 patients were younger and more often received blood or marrow transplantation (BMT). Even among patients receiving BMT, 11q23 patients had a shorter median survival (9 vs. 12-31 months for the other subgroups). However, among 11q23 patients, those receiving BMT survived longer, with 1- and 5-year survivals of 43% and 18% compared with 23% and 7% for patients not transplanted. With regard to prior therapy, 11q23 patients, compared with other patients, received radiotherapy less often as their sole therapy and chemotherapy more often. They had received VP16, methotrexate, 6MP/6TG, L-asparaginase, daunorubicin, cytarabine, and VM26 more often, likely attributed to the high frequency of AL as their primary disease. More patients in the 11q23 subgroup had received doxorubicin, except in comparison with the 21q22 subgroup; more vincristine, except in comparison with the Rare and Unique subgroups; and more prednisone, except in comparison with the Unique subgroup. Patients in the 11q23 subgroup more often received alkylating agents (AAs) (86% vs. 59-82% for the other subgroups), and topoisomerase II inhibitors (TIs) (84% vs. 49-75%), and they more often reported exposure to AAs plus TIs without radiotherapy (33% vs. 12-21%), except in comparison with the 21q22 subgroup (36%). We performed a multivariate analysis to determine whether the adverse survival of 11q23 patients compared to other Workshop patients was explained by factors other than the presence of the 11q23 abnormality. Covariates in the final model were the five cytogenetic subgroup indicators, where the 11q23 subgroup was the referent (P < 0.0001); age at t-MDS/t-AL (P = 0.0036); previous exposure to lomustine (P < 0.0001) and mitoxantrone (P = 0.0225); BMT for t-MDS/t-AL (P = 0.0006); and karyotype complexity (P = 0.0114). The risk of death for 11q23 patients relative to patients in the 21q22, inv(16), t(15;17), and Unique subgroups was significant, even after adjustment for other risk factors (relative risks 2.3, 3.6, 3.1, and 1.5, respectively; P < 0.0001 for the first three comparisons and P = 0.0125 for the last). When a multivariable model was constructed, excluding patients with AL or MDS as their primary diagnosis, the relative risk of death for 11q23 patients was significantly higher than that of all five other cytogenetic subgroups. We conclude that among t-MDS/t-AL patients with balanced aberrations, 11q23 translocations are an independent adverse risk factor. Although BMT is the current therapy of choice, new treatment is required. PMID- 11921272 TI - 21q22 balanced chromosome aberrations in therapy-related hematopoietic disorders: report from an international workshop. AB - The International Workshop on the relationship between prior therapy and balanced chromosome aberrations in therapy-related myelodysplastic syndromes (t-MDS) and therapy-related acute leukemia (t-AL) identified 79 of 511 (15.5%) patients with balanced 21q22 translocations. Patients were treated for their primary disease, including solid tumors (56%), hematologic malignancy (43%), and juvenile rheumatoid arthritis (single case), by radiation therapy (5 patients), chemotherapy (36 patients), or combined-modality therapy (38 patients). 21q translocations involved common partner chromosomes in 81% of cases: t(8;21) (n = 44; 56%), t(3;21) (n = 16; 20%), and t(16;21) (n = 4; 5%). Translocations involving 15 other partner chromosomes were also documented with involvement of AML1(CBFA2/RUNX1), identifying a total of 23 different 21q22/AML1 translocations. The data analysis was carried out on the basis of five subsets of 21q22 cases, that is, t(8;21) with and without additional aberrations, t(3;21), t(16;21), and other 21q22 translocations. Dysplastic features were present in all 21q22 cases. Therapy-related acute myeloid leukemia (t-AML) at presentation was highest in t(8;21) (82%) and lowest in t(3;21) (37.5%) patients. Cumulative drug dose exposure scores for alkylating agents (AAs) and topoisomerase II inhibitors indicated that t(3;21) patients received the most intensive therapy among the five 21q22 subsets, and the median AA score for patients with secondary chromosome 7 aberrations was double the AA score for the entire 21q22 group. All five patients who received only radiation therapy had t(8;21) t-AML. The median latency and overall survival (OS) for 21q22 patients were 39 and 14 months (mo), compared to 26 and 8 mo for 11q23 patients, 22 and 28 mo for inv(16), 69 and 7 mo for Rare recurring aberrations, and 59 and 7 mo for Unique (nonrecurring) balanced aberration (latency P < or = 0.016 for all pairwise comparisons; OS, P < or = 0.018 for all pairwise comparisons). The percentages of 21q22 patients surviving 1 year, 2 years, and 5 years were 58%, 33%, and 18%, respectively. Noticeable differences were observed in median OS between 21q22 patients (n = 7) receiving transplant (BMT) (31 mo) compared to 21q22 patients who received intensive non-BMT therapy (n = 46) (17 mo); however, this was nonsignificant because of the small sample size (log-rank, P = 0.33). t-MDS/t-AML with balanced 21q22 aberrations was associated with prior exposure to radiation, epipodophyllotoxins, and anthracyclines, dysplastic morphologic features, multiple partner chromosomes, and longer latency periods when compared to 11q23 and inv(16) t-MDS/AML Workshop subgroups. In general, patients could be divided into two prognostic risk groups, those with t(8;21) (median OS, 19 mo) and those without t(8;21) (median OS, 7 mo) leukemia (log-rank, P = 0.0007). PMID- 11921273 TI - Balanced chromosome abnormalities inv(16) and t(15;17) in therapy-related myelodysplastic syndromes and acute leukemia: report from an international workshop. AB - The Workshop identified 48 unselected patients with therapy-related myelodysplastic syndrome or acute myeloid leukemia (t-MDS/t-AML) and inv(16), and 41 patients with t(15;17) after chemotherapy (CT) and/or radiotherapy (RT) for a malignant or nonmalignant disease. The primary diseases were: breast cancer, 33 patients; lymphomas, 24 patients; various other solid tumors, 30 patients; and nonmalignant diseases, 2 patients. The general type of previous therapy was RT only in 10 patients with an inv(16) and in 12 patients with a t(15;17), alkylating agents plus topoisomerase II inhibitors in 24 patients with an inv(16) and in 18 patients with a t(15;17), topoisomerase II inhibitors only in 5 patients with an inv(16) and in 2 patients with a t(15;17), alkylating agents only in 6 patients in each subgroup, and other types of chemotherapy in 3 patients in each subgroup. Most CT-treated patients (69%) also received RT. The latency period to development of t-MDS/t-AML was short: a median of 22 months in patients with inv(16) and 29 months in patients with t(15;17). Twenty-six patients (54%) with an inv(16) and 17 patients (41%) with a t(15;17) had additional cytogenetic abnormalities, which were unrelated to age and survival in both subgroups. Trisomy of chromosomes 8, 21, and 22 and del(7q) were the most frequent additional abnormalities in the inv(16) subgroup, whereas +8, -5, and del(16q) were most frequent in the t(15;17) subgroup. The disease was overt t-AML in 38/48 patients (79%) with an inv(16) and in 38/41 patients (93%) with a t(15;17). Thirty-three of 39 intensively treated patients (85%) with an inv(16) obtained a complete remission, whereas 24 of 35 intensively treated patients (69%) with a t(15;17) obtained a complete remission. The median overall survival of intensively treated patients was 29 months in both cytogenetic subgroups. In the inv(16) subgroup, patients younger than 55 years of age had a longer survival when compared with older patients (P = 0.006). The study supports the observation that t-MDS/t-AML with inv(16) and t(15;17) is often associated with prior therapy with topoisomerase II inhibitors; however, a notable finding was the high frequency of treatment with only radiotherapy, 29% of t(15;17) and 21% of inv(16). Response rates to intensive chemotherapy in this study were comparable to those of de novo disease. PMID- 11921274 TI - Rare recurring balanced chromosome abnormalities in therapy-related myelodysplastic syndromes and acute leukemia: report from an international workshop. AB - Seventy-seven patients were identified with Rare recurring (excluding 11q23, 21q22, inv(16), and t(15;17)) chromosome abnormalities among 511 patients with treatment-related myelodysplastic syndromes and acute leukemia accepted from centers in the United States, Europe, and Japan. The abnormality subsets included 3q21q26 (17 patients), 11p15 (17 patients), t(9;22)(q34;q11) (10 patients), 12p13 (9 patients), t(8;16)(p11;p13) (9 patients), and an "other" subset, which included t(6;9)(p23;q34) (3 patients), t(10;11)(p13;q13 approximately q21) (3 patients), t(1;17)(p36;q21) (2 patients), t(8;14)(q24;q32) (2 patients), t(11;19)(q13;q13) (2 patients), t(1;3)(p36;q21) (2 patients), and t(3;5)(q21;q31) (1 patient). Increased karyotypic complexity with additional balanced and unbalanced rearrangements was observed in 70% of cases. Among 54 cases with secondary abnormalities, chromosome 5 and/or 7 abnormalities were observed in 59%. The most frequent primary diseases were breast cancer (24 cases), Hodgkin disease (14 cases), non-Hodgkin lymphoma (10 cases), and de novo ALL (5 cases). Thirty-seven patients received alkylating agents plus topoisomerase II inhibitors with or without radiation therapy. The presenting diagnosis was t-AML in 47 cases, t-MDS in 23 cases (10 progressed to t-AML), and t-ALL in seven cases, five of whom had a t(9;22). The median latency time from initiation of original therapy to therapy-related disease diagnosis was quite long (69 months), and the overall median survival from the date of therapy-related disease diagnosis was very short (7 months). The 1-year survival rate was 34 +/- 7%, with no significant differences among subsets. Comparison with previously reported cases showed increased karyotypic complexity and adult presentation of pediatric associated chromosome abnormalities. PMID- 11921275 TI - Unique balanced chromosome abnormalities in treatment-related myelodysplastic syndromes and acute myeloid leukemia: report from an international workshop. AB - A total of 123 balanced rearrangements, including 26 occurring as a sole anomaly, not known to be recurrent in myelodysplastic syndromes (MDS) or acute myeloid leukemia (AML) prior to the Workshop, were ascertained retrospectively from 104 patients with treatment-related MDS/AML (t-MDS/t-AML). Thirteen of the aberrations were reported previously in single cases and hence may be classified as recurrent as a result of the Workshop. Patients with Unique aberrations had complex karyotypes more often (P < 0.001 for all pairwise comparisons) than did other Workshop subgroups, with 72% having 3 or more aberrations. Among 85 cases with secondary chromosomal abnormalities, -5, -7, del(5q), and del(7q) were observed in 76%, which is significantly higher (P < or = 0.007 for all pairwise comparisons) than the frequencies found in the Workshop subgroups of patients with previously known recurring aberrations. The chromosome bands most often involved in balanced aberrations were 1p36 and 3q26-27. Treatment exposure was significantly different (less topoisomerase II inhibitor exposure, more radiotherapy-only exposure) than for patients with 11q23 (P < 0.001 and P = 0.002, respectively) and 21q22 (P = 0.007 and P = 0.002, respectively) abnormalities. The median time from the first toxic exposure to secondary disease, 59 months, was significantly longer (P < or = 0.016 for all significant pairwise comparisons) than the median latency of all other patients except those in the Rare subgroup, and the median survival time, 7 months, was significantly shorter than for patients in the 21q22, inv(16), and t(15;17) subgroups (P < or = 0.002 for all pairwise comparisons), but similar to patients in the 11q23 and Rare subgroups. In contrast to known recurring abnormalities, significantly more patients (61%, all P < 0.001) presented with t-MDS, with over one-third of these patients progressing to t-AML. Thus, this group of patients appears to be more similar to the typical t-MDS/t-AML patients, with complex karyotypes as well as chromosome 5 and 7 abnormalities, than to those with recurrent balanced rearrangements. PMID- 11921276 TI - Comprehensive karyotyping of the HT-29 colon adenocarcinoma cell line. AB - The tumor cell line HT-29 was derived from a primary adenocarcinoma of the rectosigmoid colon. HT-29 is hypertriploid (3n+) and has accumulated numerous chromosomal structural aberrations. To identify material involved in chromosome rearrangements, we performed a comprehensive cytogenetic analysis using G banding, spectral karyotyping (SKY), and fluorescence in situ hybridization (FISH). The combination of molecular cytogenetic techniques enabled us to define the first comprehensive karyotype for HT-29. Seventeen marker chromosomes were found in 75-100% of metaphase cells, generally in a single copy per cell. We confirmed the composition of eight previously described markers, refined the classification of seven others, and identified two novel marker chromosomes. Notable aberrations included a reciprocal translocation between chromosomes 6 and 14 and an unusual, large derivative chromosome 8 composed entirely of 8q material. The telomere status, evaluated by FISH, revealed telomeric signals at the termini of all chromosomes. No interstitial telomeric sequences were observed in any cell. Although numerous chromosomal aberrations are present in HT-29, the cell line appears to have retained a high level of genomic stability during passage in culture since undergoing transformation. The excellent resolving power of SKY, coupled with additional information obtained from molecular cytogenetic analyses, will improve our ability to identify genetic lesions characteristic of cancer. PMID- 11921277 TI - APC/CTNNB1 (beta-catenin) pathway alterations in human prostate cancers. AB - Genetic alterations serve as beacons for the involvement of specific pathways in tumorigenesis. It was previously shown that 5% of prostate tumors harbor CTNNB1 mutations, suggesting that this tumor type may involve a deregulated APC/CTNNB1 pathway. To explore this possibility further, we searched for mutations in genes implicated in this pathway in 22 samples that included cell lines, xenografts, and primary tumors. We identified seven alterations: two in CTNNB1, three in APC, and two in hTRCP1 (also known as BTRC) which controls the degradation of CTNNB1. Alterations in the CTNNB1 regulatory domain, APC, and hTRCP1 were mutually exclusive, consistent with their equivalent effects on CTNNB1 stability. These results suggest that CTNNB1 signaling plays a critical role in the development of a significant fraction of prostate cancers. Moreover, they provide the first evidence that hTRCP1 plays a role in human neoplasia. PMID- 11921278 TI - Sequencing of intron 3 of HMGA2 uncovers the existence of a novel exon. AB - Aberrations affecting the gene encoding the high mobility group protein HMGA2 (formerly HMGIC) have been found in a variety of human tumors, e.g., uterine leiomyomas, lipomas, and pulmonary chondroid hamartomas. These aberrations lead to fusion genes, transcriptional up-regulation, or aberrant transcripts of HMGA2. In the latter case, truncated transcripts consisting of exons 1 to 3 of HMGA2, encoding the three DNA-binding domains, and ectopic sequences derived from chromosome 12 are frequent. There are several lines of evidence indicating that the biological and tumorigenic features of truncated HMGA2 derivatives, i.e., those composed of the DNA-binding domains and a shortened acidic tail, clearly differ from those of the normal protein consisting of three DNA-binding domains and one large acidic tail. By sequencing the complete 112 kb third intron of HMGA2, we were able to detect several of the ectopic sequences, known as fused to HMGA2. Expression studies revealed co-expression of one of these transcripts with the normal transcript in tumors with 12q14-15 aberrations as well as in other tumors, and in normal tissues. Thus, this transcript (HMGA2b) is flanked by an alternative terminal exon of HMGA2. Due to the loss of the part encoding the acidic tail, the expression of the latter transcript may have more striking effects than the "wild type" HMGA2 (HMGA2a) in terms of tumorigenesis. This finding clearly indicates that functional studies also should address the role of the HMGA2b transcript. PMID- 11921279 TI - Mutations of the AML1 gene in acute myeloid leukemia of FAB types M0 and M7. AB - The AML1 gene encodes a transcription factor that, together with its heterodimeric partner CBFB, regulates a number of target genes that are essential for normal hemopoiesis. In acute myeloid leukemia (AML), AML1 is disrupted not only by chromosomal translocations but also by mutations in the runt domain, which binds both DNA and CBFB. Acquired mutations have been described predominantly in the AML FAB type M0. To date, most patients appear to have biallelic disease, suggesting a complete lack of normal AML1 function. Inherited loss of function mutations thought to lead to haploinsufficiency also have been described in patients who have a familial disorder with predisposition to AML (FPD/AML), indicating the role of AML1 in megakaryopoiesis. Using single-strand conformation polymorphism analysis, we studied the AML1 runt domain in 41 patients with M0 AML and identified potentially pathologic mutations in five (12%). Biallelic disease could be confirmed in only one patient, using loss of heterozygosity studies. At least three of the mutations would lead to truncated proteins similar to those reported in FPD/AML, suggesting that haploinsufficiency plays a role in the pathogenesis of this minimally differentiated type of leukemia. The incidence of acquired mutations in AML patients with acute megakaryoblastic leukemia (FAB type M7) was the same as that reported in other non-M0 patients, with only one mutation detected in 20 (5%) patients studied. PMID- 11921281 TI - Double minute chromosomes in acute myeloid leukemia and myelodysplastic syndrome: identification of new amplification regions by fluorescence in situ hybridization and spectral karyotyping. AB - Double minute chromosomes (dmin) are small chromatin bodies consisting of genes amplified in an extrachromosomal location. dmins are uncommon in hematologic malignancies; they are seen primarily in acute myeloid leukemia, with amplification of the MYC oncogene or, less frequently, the MLL transcription factor. Nine patients with hematologic malignancies with dmin were seen at the Roswell Park Cancer Institute between 1985 and 2000; eight had acute myeloid leukemia and one a myelodysplastic syndrome. Fluorescence in situ hybridization (FISH) demonstrated MYC amplification on dmin in four patients, but MLL amplification was not seen. Spectral karyotyping showed that the dmin derived from chromosome 11 in one patient and from chromosome 19 in two others without MYC or MLL amplification; derivation from these chromosomes was confirmed by FISH with chromosome paint probes. The dmin of chromosome 11 origin hybridized to a bacterial artificial chromosome (BAC) RP11-112M22 that maps to 11q24.3 and is predicted to contain ETS1 and other markers, including D11S11351 and D11S4091. The dmin of chromosome 19 origin in one patient hybridized to BACs RP11-46I12 and RP11-110J19; in the other patient, these clones did not hybridize with the dmin, but were found to be amplified on a marker chromosome that was derived from chromosome 19 in that patient's cells. These BACs have been mapped to 19q12 19q13.1 and 19q11-19q13.1, respectively, and are predicted to contain the markers D19S409 and D19S919 and the gene for ubiquinol-cytochrome C reductase, Rieske iron-sulfur polypeptide1 (UQCRFS1). dmin originating from chromosome 19 have not been reported previously in hematologic malignancies. PMID- 11921280 TI - Aberrations of the hSNF5/INI1 gene are restricted to malignant rhabdoid tumors or atypical teratoid/rhabdoid tumors in pediatric solid tumors. AB - The hSNF5/INI1 gene, which encodes a subunit of the SWI/SNF family of chromatin remodeling complexes and is located at 22q11.2, has been reported as a tumor suppressor gene inactivated in malignant rhabdoid tumors (MRTs). We analyzed this gene in varieties of pediatric solid tumors including MRTs, using the reverse transcription-polymerase chain reaction (PCR) and PCR-single strand conformation polymorphism method. We found 5 homozygous deletions, 2 truncated mutations, one missense mutation, and one silent mutation of the hSNF5/INI1 gene in 7 MRT cell lines, and one homozygous deletion, one microdeletion, one splicing acceptor site mutation, and one absence of expression in 7 fresh tumor tissues of MRT and atypical teratoid (AT)/rhabdoid tumors (RTs). Homozygous deletions were also found in one (KYM-1) of 8 rhabdomyosarcoma (RMS) cell lines. To investigate characteristics of the KYM-1 cell line, we have established KYM-1 tumors in nude mice into which KYM-1 cells were transplanted. Notably, we found that MyoD1, known as a marker for RMS, was not expressed in the KYM-1 cell line as well as MRT cell lines and fresh tumors. Histopathologic, cytogenetic, and molecular studies of the KYM-1 cell line and KYM-1 tumors in nude mice have revealed that this RMS cell line should be MRT rather than RMS. RMS-carrying aberrations of the hSNF5/INI1 gene should be reevaluated. No aberrations of this gene were found in the other 34 cell lines or 80 fresh tumor specimens except the single nucleotide polymorphisms in the 3' noncoding region. These results suggest that alterations of the hSNF5/INI1 gene were restricted to MRTs or AT/RTs in pediatric solid tumors. PMID- 11921282 TI - Combined comparative genomic hybridization and genomic microarray for detection of gene amplifications in pulmonary artery intimal sarcomas and adrenocortical tumors. AB - Identification of gene amplifications in human tumors is important for the understanding of tumorigenesis and may lead to discovery of diagnostic and prognostic markers. In this study, we used a microarray-based comparative genomic hybridization (CGH) technique, combined with conventional CGH, to identify gene amplifications in 43 tumors including eight pulmonary artery intimal sarcomas and 35 adrenocortical tumors. Conventional CGH revealed gains or amplifications of 12q13-q15 in six sarcomas and in two adrenocortical carcinomas. Using microarrays, we demonstrated that, among genes located on 12q13-q15, SAS/CDK4 were amplified in six sarcomas, and MDM2 and GLI in five and four sarcomas, respectively. The two adrenocortical tumors showed coamplifications of SAS/CDK4 and MDM2. Furthermore, PDGFRA (located on 4q12) amplification was identified in five sarcomas. Our data demonstrate: (1) amplifications of SAS/CDK4, MDM2, GLI, and PDGFRA are strongly associated with the tumorigenesis of pulmonary artery intimal sarcomas, whereas SAS/CDK4 and MDM2 coamplification may contribute to the progression of adrenocortical tumors; (2) microarray-based CGH is a useful tool for simultaneous detection of multiple gene amplifications, with a high sensitivity and resolution compared to that of conventional CGH. PMID- 11921283 TI - Comprehensive mutational analysis of the VHL gene in sporadic renal cell carcinoma: relationship to clinicopathological parameters. AB - To delineate more precisely the somatic von Hippel-Lindau disease (VHL) gene alteration as well as to elucidate its etiologic role in renal tumorigenesis, we examined a total of 240 sporadic renal cell carcinomas (RCCs) for somatic VHL gene alterations by DNA-SSCP followed by sequencing, methylation-specific PCR assay, microsatellite LOH study, and Southern blot analysis. Intragenic mutation of the VHL gene was found exclusively in clear-cell or variant-type RCCs at a frequency of 51% (104/202). Hypermethylation of the VHL promoter region was detected in an additional 11 clear-cell RCCs. Microsatellite analysis demonstrated that LOH of the VHL locus was found in 140/155 (90%) informative clear-cell RCCs. The VHL gene therefore seems to be inactivated in a two-hit manner by intragenic mutation or hypermethylation plus allelic loss in clear-cell RCC. Genomic rearrangement of the VHL gene detected by Southern analysis was not found (0/216 cases); this is in contrast to germ lines in which Southern aberrations consisted of 7-19% of the mutations. Clinicopathologic data demonstrated that VHL mutation/LOH did not vary according to tumor progression in clear-cell RCC, including tumor diameter, stage, grading, distant metastasis, and lymph node metastasis. Interestingly, VHL mutation was significantly less frequent in RCCs occurring in younger (< or = 55 years) than that in older (> or = 56 years) patients. These data suggested that the inactivation of the VHL tumor suppressor gene is a specific genetic change in clear-cell RCC, and that it may occur at an early or first step in the clear-cell tumorigenic pathway rather than as a late event. PMID- 11921285 TI - The position of pulmonary carcinoids within the spectrum of neuroendocrine tumors of the lung and other tissues. AB - Bronchopulmonary carcinoids comprise 25% of all human carcinoids. The World Health Organization divides them into typical (TC) and atypical forms (ATC), distinguished by differences in mitotic counts lower or higher than 2/2 mm(2) and the presence or absence of necrosis. The reproducibility of this classification with respect to the borderline cases with 1-2 mitotic counts/2 mm(2) has been questioned. We have analyzed 15 TCs and 20 ATCs by comparative genomic hybridization. Loss of 11q was the most frequent aberration in ATC (55%), but was observed only twice in TC (13%). Deletions of 3p were seen only in ATC (25%). Meta-analysis of our data and data from 218 neuroendocrine tumors and 50 non small-cell lung carcinomas obtained from the literature revealed differences between carcinoids and carcinomas. For example, loss of 5q is frequent in lung carcinomas (75%) but is rarely seen in carcinoids (1.4%). Deletions of 11q are less frequent in neuroendocrine lung carcinomas than in ATC. To obtain a more objective survey of the relationship of pulmonary carcinoids to other neuroendocrine tumors and lung carcinomas, we created a hierarchical clustering dendrogram. This statistical approach resulted in a clear separation of carcinoids and carcinomas, which both built up different clusters. In summary, this study demonstrates the benefit of chromosomal analysis supplementary to the diagnosis of bronchopulmonary carcinoids. We also identified the feasibility of hierarchical clustering to get some clues on relationship between different tumor types. This study further argues against a transition of ATC to high-grade neuroendocrine lung carcinoma. PMID- 11921284 TI - Gains of 12q are the most frequent genomic imbalances in adult fibrosarcoma and are correlated with a poor outcome. AB - Comparative genomic hybridization was used to analyze 41 adult fibrosarcomas from 34 patients. Thirty-one patients showed in their tumors DNA sequence copy number changes (mean 11, range 3-25). The minimal common regions for the most frequent gains were narrowed down to 12q21 (18 cases); 12q14-q15 and 14q22 (16 cases each); 4q22, 7q31, and 14q23-q24 (15 cases each); and 4q21, 4q23-q24, 8q22, and 12q22 (14 cases each). Twenty-five high-level amplifications were observed in 12 samples. 12q21 and 18p were affected three times each; and 1p21, 4q31.3, 7p21, 12q14-q15, Xp22.1-p22.2, and Xq22-q23 two times each. Losses were less frequent than gains. Early stages of adult fibrosarcomas were characterized by frequent gains of chromosomes 2, 4q, and 14q, whereas gains of chromosomes 7 and 8q were associated with progression. Gains of 12q were frequent in all of the developmental steps of this soft-tissue sarcoma. By investigation of several tumors of the same patient, a number of corresponding changes were always detected. Adult fibrosarcomas from patients who died during the observation time showed statistically significant more frequent gains of 8q, 12q, 13q, and 15q compared to the fibrosarcomas of patients who are alive. Gains and high-level amplifications of 12q14-q22, which were the most frequent genomic imbalances, partly reflected an MDM2 amplification, indicating the importance of this region in the tumorigenesis of sarcomas. In adult fibrosarcomas, a gain of 12q22 correlated significantly (P = 0.028) with a poor overall survival rate. PMID- 11921286 TI - Molecular genetic analysis of chromosome 9 candidate tumor-suppressor loci in bladder cancer cell lines. AB - Underrepresentation of chromosome 9 is a common finding in bladder cancer. Frequent loss of the whole chromosome suggests the presence of at least one relevant tumor suppressor gene on each arm. Candidate regions identified by loss of heterozygosity (LOH) analysis include a region at 9p21 containing CDKN2A, which encodes p16 and p14(ARF), a large region at 9q12-31 including PTCH and many other genes, a small region at 9q32-33, which includes the DBCCR1 gene, and a region at 9q34 including the TSC1 gene. Experimental replacement of genes or chromosomes into tumor cells with appropriate deletions or mutations represents an important approach to test the functional significance of candidate tumor suppressor genes. Loss of an entire copy of chromosome 9 in many bladder tumor cell lines provides no indication of which gene or genes are affected, and selection of appropriate recipient cells for gene replacement is difficult. We have investigated three candidate tumor suppressor genes on chromosome 9 (CDKN2A, DBCCR1, and TSC1), at the DNA level and by expression analysis in a panel of bladder tumor cell lines, many of which have probable LOH along the length of the chromosome, as indicated by homozygosity for multiple polymorphic markers. Cytogenetically, we found no reduction in the numbers of chromosomes 9 relative to total chromosome count. Homozygous deletion of the CDKN2A locus was frequent but homozygous deletion of TSC1 was not found. A new cell line, DSH1, derived from a pT1G2 transitional cell carcinoma with known homozygous deletion of DBCCR1, is described. This study identifies suitable cell lines for future functional analysis of both CDKN2A and DBCCR1. PMID- 11921287 TI - Mapping of nasopharyngeal carcinoma tumor-suppressive activity to a 1.8-megabase region of chromosome band 11q13. AB - Nasopharyngeal carcinoma (NPC) is a malignancy that is particularly prevalent among populations from Southern China and Southeast Asian countries. Evidence for a genetic contribution to the disease has been documented, although the genetic basis for NPC development is not yet fully understood. Previous functional evidence of tumor-suppressive activity on chromosome band 11q13 in NPC was obtained using a microcell-mediated chromosome-transfer approach with HONE1 NPC cells. In the present study, this region was subjected to a detailed investigation of microcell hybrids and their tumor segregants using microsatellite analysis to narrow down the region of tumor-suppressive activity. Fluorescence in situ hybridization was also performed with BAC and cosmid probes to confirm the microsatellite data. The critical region responsible for tumor suppression was narrowed down to a 1.8-Mb interval, which does not tolerate an additional normal allele by chromosome transfer. One or two alleles from either endogenous or exogenous chromosomes at 11q13 were consistently eliminated during tumor growth. Results of this study suggest that a candidate tumor-suppressor gene, not the MEN1 gene, maps between D11S4907 and GSTP1 in NPC. PMID- 11921288 TI - Identification of amplified and expressed genes in breast cancer by comparative hybridization onto microarrays of randomly selected cDNA clones. AB - Microarray analysis using sets of known human genes provides a powerful platform for identifying candidate oncogenes involved in DNA amplification events but suffers from the disadvantage that information can be gained only on genes that have been preselected for inclusion on the array. To address this issue, we have performed comparative genome hybridization (CGH) and expression analyses on microarrays of clones, randomly selected from a cDNA library, prepared from a cancer containing the DNA amplicon under investigation. Application of this approach to the BT474 breast carcinoma cell line, which contains amplicons at 20q13, 17q11-21, and 17q22-23, identified 50 amplified and expressed genes, including genes from these regions previously proposed as candidate oncogenes. When considered together with data from microarray expression profiles and Northern analyses, we were able to propose five genes as new candidate oncogenes where amplification in breast cancer cell lines was consistently associated with higher levels of RNA expression. These included the HB01 histone acetyl transferase gene at 17q22-23 and the TRAP100 gene, which encodes a thyroid hormone receptor-associated protein coactivator, at 17q11-21. The results demonstrate the utility of this microarray-based CGH approach in hunting for candidate oncogenes within DNA amplicons. PMID- 11921289 TI - Genetic analysis of mediastinal nonseminomatous germ cell tumors in children and adolescents. AB - Primary mediastinal germ cell tumors (M-GCTs) represent a heterogeneous group of tumors that varies with regard to age at presentation, histologic differentiation, and outcome. We retrospectively analyzed archival tissue samples of mediastinal mature and immature teratomas (n = 15) and malignant nonseminomatous M-GCTs (n = 20) with comparative genomic hybridization (CGH). The aim of this study was to define distinct genetic subgroups of M-GCT among the pediatric cohort that may differ in their clinical behavior and prognosis. All pure teratomas showed normal CGH profiles. Malignant M-GCTs in infants and children < 8 years old most frequently showed a gain of 1q, 3, and 20q and a loss of 1p, 4q, and 6q. Gain of 12p and sex chromosomal abnormalities were not observed in this age group. In contrast, the gain of 12p was the most common aberration in M-GCTs that arose in children > or = 8 years old. Additional recurrent changes included the loss of chromosome 13 and the gain of chromosome 21. All ten adolescents with malignant M-GCT were male, and five showed a gain of the X chromosome. In two of these five patients, Klinefelter syndrome was confirmed by cytogenetic analysis or by fluorescence in situ hybridization (FISH). In conclusion, CGH analysis of M-GCTs defines distinct genetic subgroups. Mediastinal teratomas show no genetic gains or losses. Malignant M-GCTs in children < 8 years old show the same pattern of gains and losses identified in sacrococcygeal and testicular GCTs at this age, and they lack sex-chromosomal abnormalities. Malignant M-GCTs in children > or = 8 years old show the same genetic profile previously reported in gonadal GCTs at this age. In addition, approximately 50% demonstrate a gain of the X chromosome, consistent with Klinefelter syndrome. Cooperative group studies reveal a significantly better prognosis of malignant M-GCT arising in infants compared to that in adolescents, suggesting that these genetic differences are associated with differences in clinical behavior. PMID- 11921291 TI - Genomic organization and expression of the rearranged REL proto-oncogene in the human B-cell lymphoma cell line RC-K8. AB - The human large B-cell lymphoma cell line RC-K8 has a rearranged REL locus that is transcribed into a chimeric mRNA, termed REL-NRG (Non-Rel Gene). By analyzing the recently completed human genome sequence, we have found that the normal REL and NRG loci are separated by approximately 28 megabase pairs on chromosome 2, suggesting that a deletion created the REL-NRG locus in RC-K8 cells. Using computer-based and molecular approaches, we have determined the structure of the altered REL locus in RC-K8 cells. The REL-NRG transcript is encoded by 7 REL exons and 6 NRG-derived exons. Direct DNA sequencing has identified the site of the REL-NRG fusion in RC-K8 cells. We also show that both wild-type c-Rel and c Rel-Nrg proteins are expressed and in a complex in RC-K8 cells. Furthermore, like c-Rel, c-Rel-Nrg is a cytoplasmic protein when overexpressed in fibroblasts in culture and can bind to a kappaB DNA site in vitro. PMID- 11921290 TI - Late-appearing MLL rearrangement arising as a secondary change in adult acute myeloid leukemia. AB - Rearrangements of MLL are regarded as primary oncogenic events in acute leukemia. We report the case of a patient with acute myeloid leukemia with a complex abnormal karyotype at diagnosis who showed a putative stem line with monosomy 5 and a rearrangement of chromosome 17 as the only abnormalities and an evolved clone with an additional t(7,16,11)(p1?4;p13;q23) after treatment. Fluorescence in situ hybridization analysis confirmed that the translocation had resulted in an MLL rearrangement not present in the stem line or in the complex clones found at diagnosis. To date, this is the first report of an MLL rearrangement evolving as a secondary abnormality within a preexisting leukemic clone. PMID- 11921292 TI - American Society for Laser Medicine and Surgery 22nd Annual Meeting. Atlanta, Georgia, USA, April 10-14, 2002. Abstracts. PMID- 11921302 TI - Opening the Gates for vaccine development in the real world. PMID- 11921303 TI - Metabolic co-operation between biochemically marked mammalian cells in tissue culture. 1969. PMID- 11921304 TI - Interactions of SV40 large T antigen and other viral proteins with retinoblastoma tumour suppressor. AB - Simian virus 40 large T antigen, human papilloma virus E7 and adenovirus E1A are all potent oncoproteins that can induce several types of tumours. One of the major functions of these oncoproteins is to interact with the retinoblastoma tumour suppressor protein, Rb, a master switch of the mammalian cell cycle, and to inactivate its function. Rb promotes cell-cycle arrest by recruiting and regulating proteins involved in the transcription of cell proliferation genes. The binding of viral oncoproteins to Rb disrupts the Rb-E2F complex, a central component in the Rb-mediated cell-cycle network. The crystal structures of Rb pocket-viral oncoprotein complexes indicate that the viral proteins recognise a highly conserved region in the Rb pocket through a common motif, LxCxE, and through other unique regions within each viral protein. Although the mechanism of Rb inactivation by viral proteins is not fully understood, information at the atomic level about the interactions between the Rb pocket and viral proteins is providing some insights into how viral proteins dissociate E2F from Rb and thus how they deregulate the cell cycle. PMID- 11921305 TI - The pathogenesis of febrile seizures: is there a role for specific infections? AB - Although fever is regarded as the main trigger in the pathogenesis of febrile seizures (FS), it is not supposed to be the unique causative factor. In FS, there is a strong familial predisposition. This does not exclude infections as a causative factor because subtle genetic polymorphisms have been demonstrated to affect the course of infections. We review the literature on: (1) the role of fever, especially the height of temperature, its cause, and metabolic effects induced by temperature; (2) the role of heredity; (3) the role of cytokines which play a role in the induction of fever; and (4) the role of type of infection, with emphasis on newly identified agents and improved diagnostic techniques. With modern molecular techniques such as PCR, viruses have been detected in the CSF far more often than previously thought, even in the absence of pleocytosis of the CSF. This makes it difficult to distinguish FS from acute encephalitis. FS may be caused by neuroinvasion or intracerebral activation of viruses. Further studies should focus on these options because therapeutic intervention is possible and may prevent late sequelae such as recurrent FS and subsequent epilepsy. PMID- 11921306 TI - Viruses and autoimmunity: an affair but not a marriage contract. AB - Viruses are considered as causative agents and contributors to lesion expression in autoimmune disease, notions best supported by studies in animal model systems. This review discusses relationships between virus infection and autoimmunity focusing on mechanisms by which they could induce autoreactivity. The popular idea of molecular mimicry is viewed skeptically with the reviewers taking the viewpoint that viruses contribute to autoimmunity mainly by inducing several nonspecific inflammatory events that together are sufficient to trigger autoreactivity in genetically receptive hosts. PMID- 11921307 TI - Human cytomegalovirus terminase as a target for antiviral chemotherapy. AB - Herpesviral DNA packaging is a complex process involving binding and cleavage of DNA containing the specific DNA-packaging motifs, pac1 and pac2, and packaging of the resulting unit-length genomes into preformed procapsids. This process is believed to be mediated by two packaging proteins, the terminase subunits. In the case of human cytomegalovirus the terminase consists of the proteins pUL56 and pUL89. While pUL56 (i) mediates the specific binding to pac sequences on the concatamers, (ii) provides energy for the translocation of the DNA to the procapsids and (iii) associates itself with the capsid for enabling the entry of the DNA into the procapsid, pUL89 is mainly required to effect DNA cleavage. Based on the limited efficacy of the current drugs ganciclovir, cidofovir and foscarnet, new antiviral therapeutics appear to be in demand. Inhibitors targeting pUL56 and/or pUL89 may offer an attractive alternative since mammalian cell DNA replication does not involve cleavage of concatameric DNA. Drugs targeted to terminase-like proteins should therefore be safe and highly selective. PMID- 11921308 TI - The World Health Report 2000: dialogue of the deaf? PMID- 11921309 TI - Reflections on and alternatives to WHO's fairness of financial contribution index. AB - In its 2000 World Health Report (WHR), the World Health Organization argues that a key dimension of a health system's performance is the fairness of its financing system. This paper provides a critical assessment of the index of fairness of financial contribution (FFC) proposed in the WHR. It shows that the index cannot discriminate between health financing systems that are regressive and those that are progressive, and cannot discriminate between horizontal inequity on the one hand, and progressivity and regressivity on the other. The paper compares the WHO index to an alternative and more illuminating approach developed in the income redistribution literature in the early 1990s and used in the late 1990s to study the fairness of various OECD countries' health financing systems. It ends with an illustrative empirical comparison of the two approaches using data on out-of pocket payments for health services in Vietnam for two years - 1993 and 1998. This analysis is of some interest in its own right, given the large share of health spending from out-of-pocket payments in Vietnam, and the changes in fees and drug prices over the 1990s. PMID- 11921310 TI - Testing the convergent validity of the contingent valuation and travel cost methods in valuing the benefits of health care. AB - In this study, the convergent validity of the contingent valuation method (CVM) and travel cost method (TCM) is tested by comparing estimates of the willingness to pay (WTP) for improving access to mammographic screening in rural areas of Australia. It is based on a telephone survey of 458 women in 19 towns, in which they were asked about their recent screening behaviour and their WTP to have a mobile screening unit visit their nearest town. After eliminating missing data and other non-usable responses the contingent valuation experiment and travel cost model were based on information from 372 and 319 women, respectively. Estimates of the maximum WTP for the use of mobile screening units were derived using both methods and compared. The highest mean WTP estimated using the TCM was $83.10 (95% C.I. $99.06-$68.53), which is significantly less than the estimate of $148.09 ($131.13-$166.60) using the CVM. This could be due to the CVM estimates also reflecting non-use values such as altruism, or a range of potential biases that are known to affect both methods. Further tests of validity are required in order to gain a greater understanding of the relationship between these two methods of estimating WTP. PMID- 11921311 TI - Measuring willingness-to-pay for risk reduction: an application of conjoint analysis. AB - This study applies conjoint analysis (CA) to estimate the marginal willingness-to pay (MWTP) of elderly individuals for a reduction of the risk of fracture of the femur. The good in question is a hypothetical hip protector which lowers the risk of a fracture by different amounts. Other attributes are ease of handling, wearing comfort, and out-of-pocket cost, which are traded off against risk reduction. In 500 face-to-face interviews, pensioners stated whether or not they would buy the product. Results suggest that MWTP for wearing comfort exceeds that for risk reduction. Indeed, willingness-to-pay for the product as a whole is negative, indicating that it should not be included as a mandatory benefit in health insurance. PMID- 11921312 TI - Socio-economic health inequalities in Brazil: gender and age effects. AB - This paper analyses Brazilian socio-economic inequalities in health by measuring the concentration indices for the following variables: health expectancy, self assessed health status and chronic health problems. Data used were taken from the 1996/1997 Living Standard Measurement Study (LSMS). In summary, as far as gender distinction is concerned, the results showed that up to 5 years of age the mortality rates, reported appearance of chronic health problems as well as the self-assessed health indicated that the boys were in worse health condition than the girls. After this age group there was a tendency for differences to disappear until adulthood, when the situation changed and the women consistently considered themselves less healthy. With reference to the socio-economic inequalities, one generally finds larger differences within the women's groups. The analysis for the different age groups indicated that the pro-rich inequalities increased with age. Both results were clearly proved, especially for the health expectancy variable, by adopting an adjustment of the dominance concept derived from literature on economic inequalities, consisting of comparing concentration curves. PMID- 11921313 TI - Obtaining disability weights in rural Burkina Faso using a culturally adapted visual analogue scale. AB - Burden of disease (BOD) estimates used to foster local health policy require disability weights which represent local preferences for different health states. The global burden of disease (GBD) study presumes that disability weights are universal and equal across countries and cultures, but this is questionable. This indicates the need to measure local disability weights across nations and/or cultures. We developed a culturally adapted version of the visual analogue scale (VAS) for a setting in rural Burkina Faso. Using an anthropologic approach, BOD relevant health states were translated into culturally meaningful disability scenarios. The scaling procedure was adapted using a locally relevant scale. Nine hypothetical health states were evaluated by seven panels of in total 39 lay individuals and 17 health professionals. Results show that health professionals' rankings and valuations of health states matched those of lay people to a certain extent. In comparison to that of the lay people, health professionals rated seven out of nine health states as slightly to moderately less severe. The instrument scored well on inter-panel and test-retest reliability and construct validity. Our research shows the feasibility of eliciting disability weights in a rural African setting using a culturally adapted VAS. Moreover, the results of the present study suggest that it might be possible to use health professionals' preferences on disability weights as a proxy for lay people's preferences. PMID- 11921314 TI - How profitable is risk selection? A comparison of four risk adjustment models. AB - To mitigate selection triggered by capitation payments, risk-adjustment models bring capitation payments closer on average to individuals' expected expenditure. We examine the maximum potential profit that plans could hypothetically gain by using their own private information to select low-cost enrollees when payments are made using four commonly used risk adjustment models. Simulations using a privately insured sample suggest that risk selection profits remain substantial. The magnitude of potential profit varies according to the risk adjustment model and the private information plans can employ to identify profitable enrollees. PMID- 11921315 TI - Is there a kink in consumers' threshold value for cost-effectiveness in health care? AB - BACKGROUND: A reproducible observation is that consumers' willingness-to-accept (WTA) monetary compensation to forgo a program is greater than their stated willingness-to-pay (WTP) for the same benefit. Several explanations exist, including the psychological principle that the utility of losses weighs heavier than gains. We sought to quantify the WTP-WTA disparity from published literature and explore implications for cost-effectiveness analysis accept-reject thresholds in the south-west quadrant of the cost-effectiveness plane (less effect, less cost). METHODS: We reviewed published studies (health and non-health) to estimate the ratio of WTA to WTP for the same program benefit for each study and to determine if WTA is consistently greater than WTP in the literature. RESULTS: WTA/WTP ratios were greater than unity for every study we reviewed. The ratios ranged from 3.2 to 89.4 for environmental studies (n=7), 1.9 to 6.4 for health care studies (n=2), 1.1 to 3.6 for safety studies (n=4) and 1.3 to 2.6 for experimental studies (n=7). CONCLUSIONS: Given that WTA is greater than WTP based on individual preferences, should not societal preferences used to determine cost effectiveness thresholds reflect this disparity? Current convention in cost effectiveness analysis is that any given accept-rejection criterion (e.g. $50 k/QALY gained) is symmetric - a straight line through the origin of the cost effectiveness plane. The WTA-WTP evidence suggests a downward 'kink' through the origin for the south-west quadrant, such that the 'selling price' of a QALY is greater than the 'buying price'. The possibility of 'kinky cost-effectiveness' decision rules and the size of the kink merits further exploration. PMID- 11921316 TI - On being NICE in the UK: guidelines for technology appraisal for the NHS in England and Wales. PMID- 11921317 TI - Neonatal health care costs related to smoking during pregnancy. AB - RESEARCH OBJECTIVE: Much of the work on estimating health care costs attributable to smoking has failed to capture the effects and related costs of smoking during pregnancy. The goal of this study is to use data on smoking behavior, birth outcomes and resource utilization to estimate neonatal costs attributable to maternal smoking during pregnancy. STUDY DESIGN: We use 1995 data from the Center for Disease Control's (CDC) Pregnancy Risk Assessment Monitoring System (PRAMS) database. The PRAMS collects representative samples of births from 13 states (Alabama, Alaska, California, Florida, Georgia, Indiana, Maine, Michigan, New York (excluding New York City), Oklahoma, South Carolina, Washington, and West Virginia), and the District of Columbia. The 1995 PRAMS sample is approximately 25 000. Multivariate analysis is used to estimate the relationship of smoking to probability of admission to an NICU and, separately, the length of stay for those admitted or not admitted to an NICU. Neonatal costs are predicted for infants 'as is' and 'as if' their mother did not smoke. The difference between these constitutes smoking attributable neonatal costs; this divided by total neonatal costs constitutes the smoking attributable fraction (SAF). We use data from the MarketScantrade mark database of the MedStattrade mark Corporation to attach average dollar amounts to NICU and non-NICU nursery nights and data from the 1997 birth certificates to extrapolate the SAFs and attributable expenses to all states. PRINCIPAL FINDINGS: The analysis showed that maternal smoking increased the relative risk of admission to an NICU by almost 20%. For infants admitted to the NICU, maternal smoking increased length of stay while for non- NICU infants it appeared to lower it. Over all births, however, smoking increased infant length of stay by 1.1%. NICU infants cost $2496 per night while in the NICU and $1796 while in a regular nursery compared to only $748 for non-NICU infants. The combination of the increased NICU use, longer stays and higher costs result in a positive smoking attributable fraction (SAF) for neonatal costs. The SAF across all states is 2.2%. Across the states, the SAF varied from a low of 1.3% in Texas to a high of 4.6% in Indiana. CONCLUSIONS: These results further confirm the adverse effects of smoking. Among mothers who smoke, smoking adds over $700 in neonatal costs. The smoking attributable neonatal costs in the US represent almost $367 million in 1996 dollars; these costs vary from less than a million in smaller states to over $35 million in California. These costs are highly preventable since the adverse effects of maternal smoking occur in the short-run and can be avoided by even a temporary cessation of maternal smoking. These cost estimates can be used by managed care plans, state and local public health officials and others to evaluate alternative smoking cessation programs. PMID- 11921318 TI - On the empirical association between poor health and low socioeconomic status at old age. AB - Epidemiologic studies using mortality rates as indicators of health fail to find any meaningful association between poor health and low socioeconomic status in older age-groups, whereas economic studies using self-assessed health consistently find a significant positive correlation, even after controlling for self-reporting errors. Such contradictory results have not been reported for working age individuals. A simple explanation might be that the elderly samples on which the epidemiologic and economic studies are based come from different populations. However, this paper shows that similar contradictory results are obtained even when the same samples are used, simply by switching between self assessed health and mortality as health indicators. An alternative explanation is proposed, namely that these health indicators yield different results because they relate to different ranges of the latent health variable at old age. PMID- 11921319 TI - A general model of the impact of absenteeism on employers and employees. AB - Most studies on the indirect costs of an illness and the cost effectiveness of a medical intervention or employer-sponsored wellness program assume that the value of reducing the number of days employees miss from work due to illness is the wage rate. This paper presents a general model to examine the magnitude and incidence of costs associated with absenteeism under alternative assumptions regarding the size of the firm, the production function, the nature of the firm's product, and the competitiveness of the labor market. We conclude that the cost of lost work time can be substantially higher than the wage when perfect substitutes are not available to replace absent workers and there is team production or a penalty associated with not meeting an output target. In the long run, workers are likely to bear much of the incidence of the costs associated with absenteeism, and therefore be the likely beneficiaries of any reduction in absenteeism. PMID- 11921320 TI - Estimating survival gain for economic evaluations with survival time as principal endpoint: a cost-effectiveness analysis of adding early hormonal therapy to radiotherapy in patients with locally advanced prostate cancer. AB - The problem of estimating expected outcomes for the economic evaluation of treatments for which the outcome of principal interest is (quality adjusted) survival time has so far not received sufficient attention in the literature. The best estimate of expected survival is mean survival time, but with censored survival data, the true survival time for all the subjects is not known, so the mean is not defined.A possible solution to this estimation problem is illustrated by a retrospective cost-effectiveness analysis of the addition of hormonal therapy to standard radiotherapy for patients with locally advanced prostate cancer. A recently proposed method is used to approach the problem caused by censored cost data, and the impact of uncertainty is assessed by bootstrap resampling techniques. Mean survival time is estimated by a restricted means analysis with the time point of restriction determined by statistical criteria. When average total costs and mean survival time is evaluated at this time point of restriction, the result is that the combined therapy (radiotherapy plus hormonal therapy) increases mean survival time by about 1 year, while reducing the costs per patient for the French health insurance system by 12 700 FF. The time point of restriction may also be determined by other criteria and mean survival time may be estimated by extrapolating the survival curves by means of various parametric survival distributions. We show that the exact results of the economic evaluation are decisively determined by the restriction time point chosen and the approach taken to estimate mean survival time. PMID- 11921322 TI - Morality and the limits of societal values in health care allocation. AB - In this paper, we consider whether there is a clear moral justification for the proposal that societal value preferences (SVPs) should be included in Cost Effectiveness Analyses (CEA) of health care resource allocations. We argue, first, that proponents of the use of SVPs need to be clear about the relationship between these values and moral principles. In particular, once moral principles are accepted as ruling out some SVPs (such as those that are irrational or revealing prejudice), an account is required of why we need to appeal to SVPs rather than moral principles to determine a just division of health care resources. Secondly, we consider whether an independent moral justification might underwrite the use of SVPs. In various places in the literature the notions of representation, presumed consent and democratic decision making appear to be invoked as candidates for fulfilling this justificatory role. We discuss some problems with each of these justifications in the hope of eliciting a more comprehensive proposal from the proponents of SVPs. We conclude that, although a number of interesting proposals have been made, no compelling justification for including SVPs in CEA has yet been systematically articulated. PMID- 11921321 TI - Statistical determination of cost-effectiveness frontier based on net health benefits. AB - Statistical methods are given for producing a cost-effectiveness frontier for an arbitrary number of programs. In the deterministic case, the net health benefit (NHB) decision rule is optimal; the rule funds the program with the largest positive NHB at each lambda, the amount a decision-maker is willing to pay for an additional unit of effectiveness. For bivariate normally distributed cost and effectiveness variables and a specified lambda, a statistical procedure is presented, based on the method of constrained multiple comparisons with the best (CMCB), for determining the program with the largest NHB. A one-tailed t test is used to determine if the NHB is positive. To obtain a statistical frontier in the lambda-NHB plane, we develop a method to produce the region in which each program has the largest NHB, by pivoting a CMCB confidence interval. A one-sided version of Fieller's theorem is used to determine the region where the NHB of each program is positive. At each lambda, the pointwise error rate is bounded by a prespecified alpha. Upper bounds on the familywise error rate, the probability of an error at any value of lambda, are given. The methods are applied to a hypothetical clinical trial of antipsychotic agents. PMID- 11921323 TI - The familywise error rate of a simultaneous confidence band for the incremental net health benefit. AB - Interest in the use of net health benefit in cost-effectiveness analysis derives from its optimality property for decision-making. A description of the results of an economic evaluation of health care interventions is incomplete if it does not include point and interval estimates of this outcome measure. A simultaneous confidence band for the incremental net health benefit, INHB(lambda), for all lambda may be obtained by forming a confidence interval based on student's t statistic, and letting the willingness-to-pay value, lambda, run over all values. The familywise error rate (FWER) of the simultaneous confidence band is the probability that the confidence interval does not cover the true INHB(lambda) for some value of lambda. We show that the FWER equals P(T(2)>t(2)), where T(2) follows Hotelling's central distribution and that the simultaneous confidence band does not cover the true INHB(lambda) if and only if a T(2) based confidence ellipsoid does not cover the true mean c-e vector. PMID- 11921324 TI - Surviving and thriving? PMID- 11921325 TI - The first year after breast cancer diagnosis: hope and coping strategies as predictors of adjustment. AB - Given the marked individual variability in psychological adjustment in response to breast cancer, it is important to specify factors contributing to adaptive survivorship. This longitudinal study of 70 women with Stage I or II breast cancer tested the ability of situation-specific coping strategies and a more stable attribute, hope, to predict adjustment prospectively from the point shortly following diagnosis through the first year. Consonant with previous studies, coping through active acceptance at diagnosis predicted more positive adjustment across time, and avoidance-oriented coping predicted greater fear of cancer recurrence, over and above participant age and initial status on dependent variables. The hypothesis that coping through turning to religion would be more effective for less hopeful women was supported, and mixed support emerged for the hypothesis that approach-oriented coping strategies would yield greater adaptational benefits for women high in hope. Findings suggest that risk and protective factors for adjustment across the first year of survivorship can be identified even prior to definitive surgery for breast cancer, particularly when both dispositional characteristics such as hope and situation-specific coping strategies are considered. PMID- 11921326 TI - Ethnic differences in quality of life among early breast and prostate cancer survivors. AB - This study assessed quality of life (QOL) in recently diagnosed breast and prostate cancer patients of European American and Asian Pacific Islanders (specifically, individuals of Filipino, Japanese, and Native Hawaiian ancestry) to investigate whether QOL varied according to ethnicity and the relative importance of ethnicity as a predictor of QOL. Participants were identified through consecutive registrations on the Hawaii Tumor Registry, based on a diagnosis of breast or prostate cancer 4-6 months previously. QOL was measured by the QLQ-C30, a standardized questionnaire widely used in cancer patient populations. Two hundred and twenty-seven individuals participated (101 prostate and 126 breast cancer patients). QOL was similar across ethnic groups in most areas of QOL. However, differences were found in several areas, all in the direction of Filipino patients reporting worse outcomes. Hierarchical stepwise regressions supported the importance of ethnicity, controlling for clinical and demographic predictors. We conclude that additional research is needed to understand the relationship between ethnicity and QOL in cancer survivors. PMID- 11921327 TI - Life after cancer: adaptation issues faced by Chinese gynaecological cancer survivors in Hong Kong. AB - An explorative study using qualitative data collection methods was used to look at the adaptation issues faced by Chinese survivors of gynaecological cancers. Eighteen women who were free from illness and between 2 and 16 years post diagnosis participated. Content analysis of the data revealed that cancer and its treatment led to a positive revival of these women as they re-evaluated their life, started treasuring family relationships, improved their mood and became more sensitive to other people's needs. However, they also described problems with marital relationships, concerns about the future of their children, sexuality and femininity, loss of fertility and discomfort due to pain and weakness. A third of the sample reported that their husbands had extramarital affairs. Despite this, women, especially those who had good communication with their partners, commented that they maintained a loving relationship. Cultural beliefs and misconceptions about sexual functioning were also depicted as affecting adaptation to illness. Long-term psychosocial care may help these women to adapt better to the demands of the illness, especially sexual and psychosexual rehabilitation, as sexual problems were reported by all women in the study. PMID- 11921328 TI - Factors reported to influence the return to work of cancer survivors: a literature review. AB - An overview is provided of research into the return to work of cancer survivors, examining both the rate of return to work and factors impacting this return. A series of literature searches was conducted on MEDLINE and PSYCLIT databases for the years 1985-1999. Studies had to focus on the patient's perspective and had to include either the percentage of return to work or factors associated with return to work. Case studies and studies of cancer as an occupational disease were excluded. The search identified 14 studies. The mean rate of return to work was 62% (range 30-93%). The following factors were negatively associated with return to work: a non-supportive work environment, manual labour, and having head and neck cancer. Sociodemographic characteristics were not associated with return to work. For increasing age, associations were mixed. The increased survival rate of cancer patients warrants attention to the problems survivors may encounter upon their return to work. More systematic research is needed to establish more clearly the relative importance of factors associated with return to work of cancer survivors, which, in turn, would contribute to an increase in the labour participation of cancer survivors. PMID- 11921329 TI - Quality of life in childhood cancer survivors. AB - The successful treatment for children with cancer has greatly increased the survival rates for these young people compared to children diagnosed with cancer 30 years ago. These new medical realities direct attention to the psychosocial consequences of successful treatment and subsequent survival. In this paper, quality of life in 176 childhood cancer survivors (age 16-28) is assessed using a survey instrument designed for cancer survivors. In addition, the instrument is evaluated for its utility with this population. Survivors indicate that symptoms often associated with treatment are at a minimum but that other long-term effects like fatigue, aches, and pain negatively impact quality of life. They rate themselves high on happiness, feeling useful, life satisfaction and their ability to cope as a result of having had cancer but their hopefulness is tempered by uncertainty. Whereas the salience of spiritual and religious activities appears to be low, having a sense of purpose in life and perceiving positive changes as a result of cancer are associated with positive quality of life. A lower valence of physical concerns reflects the vitality and positive life outlook of a young population. PMID- 11921331 TI - Five years later: a cross-sectional comparison of breast cancer survivors with healthy women. AB - Although a number of studies have focused on initial adjustment to cancer, less is known about long-term survival issues. The present study compared breast cancer survivors with age-matched healthy controls (N=328) in terms of more subtle indicators of psychological well-being as well as their general quality of life 5 yrs post-diagnosis. Results indicated survivors generally perceive the world as less controllable and more random compared to healthy women. However, survivors perceive the same control over their daily lives as healthy women. Beliefs about personal control are most strongly associated with quality of life in both groups of women. Survivors also indicated that they derived some benefits from their experience with cancer, but these benefits had only a modest impact on quality of life. However, the belief that the experience had lasting harmful effects was associated with poor quality of life for survivors. In both groups, a continued search for meaning in life had a negative impact on quality of life. The strongest and most consistent correlate of quality of life for both survivors and healthy women was having a sense of purpose in life. PMID- 11921330 TI - Resilience, reflection, and residual stress in ovarian cancer survivorship: a gynecologic oncology group study. AB - Ovarian cancer is a life-threatening diagnosis which poses multiple challenges. The purpose of this study is to describe the quality of life (QOL) concerns and survivorship sequelae of long-term (>5 yr) early-stage ovarian cancer survivors accrued through the clinical cooperative Gynecologic Oncology Group. Forty-nine ovarian cancer survivors with a mean age at diagnosis of 55.9 yr (range 30-76) completed a telephone interview assessing QOL, psychosocial status, sexual functioning and late-effects of treatment. Results indicate that this disease free early-stage sample enjoys a good QOL, with physical, emotional, and social well-being comparable to other survivors and same-aged noncancer cohorts. However, 20% of survivors indicated the presence of long-term treatment side effects, with a subset reporting problems related to abdominal and gynecologic symptoms, and neurotoxicity. Spiritual well-being was significantly positively associated with personal growth and mental health, and negatively associated with a declining health status. Lingering psychological survivorship sequelae included fear of follow-up diagnostic tests and fear of recurrence. Forty-three percent of respondents expressed that they would likely participate in a counseling program today to discuss psychosocial issues raised by having had ovarian cancer, and 56% stated that they would have attended a support program during the initial treatment if it had been offered. This information provides some insight into the complex survivorship relationships between quality of life, long-term physical and sexual sequelae, and factors of resilience and growth which appear to promote a sense of well-being as a result of the cancer experience. PMID- 11921332 TI - Survivorship and discourses of identity. AB - Personal identity is self-evidently important to us all. Identity is a philosophically complex subject, but there is some agreement that memory, embodiment and continuity are essential components. The sense of memory includes 'future memory', the kind of memory we would like to construct for ourselves as our lives proceed. While the sense of personal identity is internal to the individual, a sense of that person's identity exists in the minds of others. Extreme experiences threaten the element of continuity, because they may bring bodily changes as well as cognitive changes that challenge central values. Restoring or preserving continuity is a major task for survivors. The ways in which people experience discontinuity because of cancer illness, and the ways in which they manage this experience emerges from the narratives of the survivors of cancer and in the narratives of health care workers who look after them. People manage discontinuity by reference to stable 'anchor points' in their beliefs and values; by re-constructing versions of their pre-experience identities, drawing on past memory and finding ways to preserve a continuity between past memory, present experience and constructions of the future; by using the experience to develop established facets of identity; and by imbuing the experience with meaning and recognising the enlarged identity made possible by survival. Those who cannot achieve a sense of continuity may feel alienated from themselves, their friends and family. All these methods of management may be used by one person to negotiate the post-experience identity in its different social interactions. The experience of the survivor can be further understood by recognising the challenge posed by extreme experience to the sense of continuity of both embodied self and memory. A satisfactory discourse of survival has yet to enter the public domain. This lack adds to the burdens of survivors, including those who have survived cancer. PMID- 11921334 TI - Development of the zebrafish inner ear. AB - Abstract Recent years have seen a renaissance of investigation into the mechanisms of inner ear development. Genetic analysis of zebrafish has contributed significantly to this endeavour, with several dramatic advances reported over the past year or two. Here, we review the major findings from recent work in zebrafish. Several cellular and molecular mechanisms have been elucidated, including the signaling pathways controlling induction of the otic placode, morphogenesis and patterning of the otic vesicle, and elaboration of functional attributes of inner ear. PMID- 11921335 TI - Adipose tissue: a vital in vivo role in mammary gland development but not differentiation. AB - Development and differentiation of the mammary gland occurs by means of critical stromal-epithelial interactions. Although many studies have attempted to understand these complex interactions, it has been difficult to demonstrate the essential role of adipose tissue in the development and function of the mammary gland. By using the A-ZIP/F-1 transgenic mice lacking in white adipose tissue (WAT), we have studied the role of adipocytes in mammary gland development and differentiation. In the absence of WAT, rudimentary mammary anlagen form but are unable to grow and branch normally, resulting in a few, short, severely distended ducts. However, during pregnancy, a tremendous amount of epithelial cell division and alveolar cell formation occurs even in the absence of adipocytes, illustrating that adipose tissue is not required for mammary gland differentiation. Mammary gland transplantation revealed that epithelial cells from these transgenic mice possess the potential for normal growth and differentiation when placed into a normal stromal environment. These experiments clearly demonstrate that the absence of adipocytes in the mammary gland results in disruption of stromal-epithelial interactions that prevent normal mammary gland development. The rudimentary epithelial anlage, however, contain mammary stem cells, which are fully capable of alveolar differentiation. PMID- 11921336 TI - Initiation of apoptosis in the developing avian outflow tract myocardium. AB - Apoptosis occurs within the cardiac outflow tract (OFT) myocardium during normal development of chick hearts. This peak of apoptosis occurs at stage 30-31 and coincides with dramatic remodeling of the OFT, suggesting that apoptosis occurs to allow proper alignment of the great vessels over their respective ventricles. The signals that initiate apoptosis in this setting are unknown. The aim of this study was to characterize the cells undergoing apoptosis in the cardiac OFT myocardium and the cells that may influence this process. Two cell populations that may initiate apoptosis of the cardiomyocytes are the cardiac neural crest (CNC) cells and epicardial cells. We examined stage 30-31 chick embryos that had undergone removal of the CNC cells or had delayed epicardial growth for alterations of apoptosis. Removal of the CNC cells did not reduce the levels or pattern of apoptosis in the OFT myocardium. In contrast, impeding the growth of the epicardium over the OFT resulted in a 57% reduction in apoptotic cells in the OFT myocardium. Analysis of the apoptotic cells within the OFT myocardium showed that as many as 92% of them expressed cardiomyocyte markers. In the quail, the endothelial marker QH1 identified a component from the epicardium, endothelial cells, in regions where apoptosis is elevated in the OFT myocardium. These results suggest that a component from the epicardium, possibly endothelial cells, is required for the initiation of apoptosis in OFT cardiomyocytes. PMID- 11921337 TI - Taste bud development in the zebrafish, Danio rerio. AB - Taste buds are chemosensory endorgans consisting of modified epithelial cells. Fish and other vertebrates use their taste bud cells to sample potential food, either selecting or rejecting substances according to their edibility. The adult gustatory system in fish has been studied thoroughly, including regeneration experiments. Taste buds occur in the epithelia of the lips, the mouth cavity, the oropharyngeal cavity, and also in the skin of the barbels, the head, and sometimes even all over the body surface. Despite its importance for feeding, little is known about the ontogeny of the fish taste system. We examined the development of taste buds in the zebrafish on the light microscopical and the scanning and transmission electron microscopical levels. Taste buds develop later than the olfactory organ and the solitary chemosensory cells, two other chemosensory systems in aquatic vertebrates. The first few taste bud primordia are visible within the epithelia of lips and gill arches 3 to 4 days after fertilization, and the first few taste buds with open receptor areas appear on the lips and simultaneously on the gill arches 4-5 days after fertilization, which coincides with the onset of feeding. Taste buds in the mouth cavity, on the head, and on the barbels are formed later in development. As seen in other fish, zebrafish taste buds contain elongate dark and light cells, termed according to their electron density. Dark cells with a cell apex of many short microvilli appear first, followed by the light cells with one large microvillus. In addition, the zebrafish has a third fusiform cell type, which appears last. This cell type is low in electron density and has a brush-like apical ending with several small microvilli. This cell type has not been described previously. Furthermore, in zebrafish, the ontogenetic processes of taste bud formation differ from regenerative processes described in the literature. PMID- 11921338 TI - Cell-matrix interactions during development and apoptosis of the mouse mammary gland in vivo. AB - Epithelial cell survival is dependent on extracellular signals provided by both soluble factors and by adhesion. In the mammary gland, extensive apoptosis of epithelial cells occurs rapidly when lactation ceases, but the mechanism of apoptosis induction is not known. In tissue culture, mammary epithelial cells require laminin as a survival ligand and specific beta1 integrins are necessary to suppress apoptosis. To explore the possibility that dynamic changes in cell matrix interactions contribute to the onset of apoptosis during mammary involution in vivo, a detailed immunohistochemical analysis of the expression of integrin subunits and their extracellular matrix ligands during mouse mammary gland development has been performed. The kinetics of apoptosis were determined by using tissue samples obtained from virgin, pregnant, lactating, and involuting gland. The maximal elevation of apoptosis occurred within 24 hr of weaning as determined by histologic analysis and caspase-3 staining. A wide variety of laminin subunits, together with nidogen-1 and -2, and perlecan were identified within the basement membrane region of epithelial ducts, lobules, and alveoli in both human and mouse mammary gland. However, no change in the distribution of any of the basement membrane proteins or their cognate integrin receptors was observed during the transition from lactation to apoptosis. Instead, we discovered that altered ligand-binding conformation of the beta1 integrin to a nonbinding state coincided with the immediate onset of mammary apoptosis. This finding may provide a novel dynamic mechanism for inhibiting the transduction of extracellular matrix survival signals, thereby contributing to the onset of apoptosis in a developmental context in vivo. PMID- 11921339 TI - Vestibular dysgenesis in mice lacking Abr and Bcr Cdc42/RacGAPs. AB - The inner ear develops from a simple epithelium (otic placode) into the complex structures specialized for balance (vestibule) and sound (cochlea) detection. Abnormal vestibular and cochlear development is associated with many birth defects. During recent years, considerable progress has been made in understanding the molecular bases of these conditions. To determine the biological function of two closely related GTPase activating proteins for the Cdc42/Rac GTPases, Abr and Bcr, we generated a mouse strain deficient in both of these proteins. Double null mutant mice exhibit hyperactivity, persistent circling, and are unable to swim. These phenotypes are typically found in mice with vestibular defects. Indeed, adult double null mutants display abnormal dysmorphic structures of both the saccule and utricle. Moreover, a total loss of otoconia can be seen in the utricle, whereas in the saccule, otoconia are either missing or their number is drastically decreased and they are abnormally large. Interestingly, both the cochlea and semicircular canals are normal and the double null mutant mice are not deaf. These data demonstrate that Abr and Bcr play important complementary roles during vestibular morphogenesis and that a function of Cdc42/RacGAPs and, therefore, that of the small Rho-related GTPases is critically important for balance and motor coordination. PMID- 11921340 TI - Thyroid hormone regulation of a transcriptional coactivator in Xenopus laevis: implication for a role in postembryonic tissue remodeling. AB - Thyroid hormone (TH) affects biological processes by regulating gene transcription through TH receptors (TRs). In the presence of TH, TR activates target gene transcription by recruiting one or more transcription coactivators belonging to diverse groups. Here, we demonstrate that during TH-dependent anuran metamorphosis, one such coactivator gene, the Xenopus laevis homolog of human Trip7, is up-regulated by TH. Kinetic studies suggest that Xenopus Trip7 is most likely induced indirectly by TH in a tissue-dependent manner. In the intestine, which undergoes extensive remodeling as the animal changes from being herbivorous to carnivorous, Trip7 is expressed at high levels during but not before or after metamorphosis. It is also up-regulated in other growing or remodeling tissues such as the brain and limb but not in degenerating tadpole tail skin. By using frog oocyte as a model, we show that Trip7 influences basal transcription in a chromatin structure-dependent manner but enhances the function of liganded TR regardless of the chromatin structure of the target promoter. In vitro studies indicate that Trip7 interacts directly with TR. These results suggest that during Xenopus metamorphosis, TH up-regulates, albeit indirectly, Trip7 to enhance TR function during larval-to-adult tissue transformation. PMID- 11921341 TI - Atlas of the developing inner ear in zebrafish. AB - This report provides a description of the normal developing inner ear of the zebrafish, Danio rerio, with special focus on the pars inferior. Zebrafish specimens, ranging in age from 3 to 30 days postfertilization (dpf), were processed for standard histologic sections or with a paint-fill method to show three-dimensional morphogenesis of the membranous labyrinth. Adult zebrafish (age 2 years) were also processed for inner ear paint-fills. Although development of the semicircular canals occurs rapidly (by 3 dpf), the pars inferior develops more gradually during days 5-20 postfertilization. A rudimentary endolymphatic duct emerges by 8 dpf. Differentiated hair cells of the lagenar macula are evident by 15 dpf, in a chamber located lateral and posterior to the saccule. By 20 dpf, the saccule itself is separated from the utricle, but remains connected by means of the utriculosaccular foramen. The maculae neglectae, each with differentiated hair cells, lie on the floor of the utricle near this foramen. A medial connection between the sacculi of right and left ears, the transverse canal, is also complete by 20 dpf. A ridge of mesenchyme, previously undescribed, bisects the saccule in zebrafish fry at 20-30 dpf. The images in the paint-fill atlas should provide a baseline for future studies of mutant zebrafish ears. PMID- 11921342 TI - Segment-specific expression of connexin31 in the embryonic hindbrain is regulated by Krox20. AB - Communication and interaction between cells has been shown to be important during the embryonic development of the vertebrate hindbrain, which becomes transiently subdivided into segments called rhombomeres (r). One gene family allowing intercellular communication and possibly being involved in the control of hindbrain development is the connexin family encoding gap junction channels. Here, we have characterized in detail the previously observed (Dahl et al., 1997) expression of one particular connexin gene, connexin31 (Cx31), in the mouse embryonic hindbrain and compared it with that of Cx43 and Cx36. We found transient Cx31 expression from approximately embryonic day (E) E8-E11 in two small lateral/dorsal subgroups of cells in the hindbrain. We could show that these spots of expression corresponded to r3 and r5 and that Cx31 expression in r3 and r5 was controlled by the transcription factor Krox20. In contrast, expression of Cx43 and Cx36 started later (from E9.5 and E10.5, respectively) and was confined to longitudinal stripes of expression. In addition, from E10.5 E11.5, Cx31 was expressed by a column of cells in ventral r4, most likely representing contralateral vestibulo-acoustic efferent neurons, immediately anterior to a ventral column expressing Cx36 at the same stage. From E11.5 onward, another site of Cx31 expression was detected in the boundary cap cells in the entry/exit points of all mixed sensory/motor and in the entry points of pure sensory nerves. This expression was not present in the boundary cap cells of the exit points of pure motor nerves. So far, our analysis of the hindbrain area of Cx31-deficient embryos in terms of projections of sensory or motor neurons or in the generation or migration of neurons has not yet revealed any obvious defects. PMID- 11921343 TI - Localization of metals in cells of pterygote insects. AB - Insects maintain the equilibrium of their internal milieu by diffusion mechanisms, but more frequently by a substantial storage of metals in the cells of numerous organs: digestive tract, Malpighian tubules, fat body, integument, and genital organs. This storage implies the precipitation, of numerous cations (Ca, Mg, K, Mn, Fe, Zn, Cu) in a structure called the spherocrystal, which originates from the endoplasmic reticulum-Golgi complex: elements precipitate on a glycosaminoglycan nucleus in thin peripheral strata. Some spherocrystals contain exclusively mineral compounds, frequently phosphates, whereas others may contain organic compounds such as urates. In some species mineralized lysosomes store Ca, Fe, Zn, and Cu. When fed additional metals found in the environment, insects such as cockroach and ant are able to stay alive and to trap the metals (Cd or Pb, for example) in the peripheral strata of spherocrystals; the cytoplasm is not altered. It seems that these insects are able to resist exposures to high levels of toxic metals. The lysosomes are able to retain toxic heavy metals (Cd or Hg, for example) within metallothionein-like proteins. PMID- 11921344 TI - Vanadocytes, cells hold the key to resolving the highly selective accumulation and reduction of vanadium in ascidians. AB - Since Henze discovered vanadium in the blood (or coelomic) cells of an ascidian in 1911, this unusual phenomenon has attracted the interest of many investigators. The highest concentration of vanadium (350 mM) in the blood cells of Ascidia gemmata, which belongs to the suborder Phlebobranchia, is 10(7) times higher than that in seawater. Of the approximately 10 types of blood cells, a combination of cell fractionation and neutron-activation analysis revealed that the signet ring cells were the true vanadocytes. In the vanadocytes, 97.6% of the vanadium is in the +3 oxidation state (III). The extremely low pH of 1.9 found in vanadocytes suggests that protons, concentrated by an H(+)-ATPase, might be linked to the accumulation of vanadium energetically. The antigen recognized by a monoclonal antibody, S4D5, prepared to identify vanadocytes, was determined to be 6-PGDH in the pentose phosphate pathway. NADPH produced in the pentose phosphate pathway in vanadocytes is thought to participate in the reduction of vanadium(V) to vanadium(IV). During embryogenesis, a vanadocyte-specific antigen first appears in the body wall at the same time that significant accumulations of vanadium become apparent. Three different vanadium-associated proteins (VAPs) were extracted from the blood cells of vanadium-rich ascidians. These are 12.5, 15, and 16 kDa in size and are associated with vanadium in an approximate ratio of 1:16. The cDNA encoding the 12.5 and 15 kDa VAPs was isolated and the proteins encoded were found to be novel. Further biochemical and biophysical characterization of the VAPs is in progress. PMID- 11921345 TI - Histochemical and ultrahistochemical localization of heavy metals in calf organs. AB - Cadmium, zinc, selenium, and copper were administered, singly or in combination, orally or subcutaneously. Experiment I included 32 calves of both sexes; six received Cd (two groups), Zn, Cd, and Zn, and Cd and Se (two groups) and one group was a control. In Experiment II (21 bulls), three were given Cd, Cd, and Cu, and Cd and Zn, respectively, and one group was a control. For light microscopy, in Experiment I the highest amounts of silver granules were present in the samples of liver, small intestine, and vesicular gland of all the exposed groups; in Experiment II the most affected organs were liver, kidney, and small intestine. For electron microscopy, in Experiment I, after administration of Cd and Zn, the highest amounts of granules were seen in the cytoplasm of hepatocytes and cells of the proximal and distal renal tubules and the lowest amounts were found in glandular cells of the pancreas. Administration of Cd and Se resulted in the presence of large numbers of granules in the nuclei and nucleoli of spermatogonies. In Experiment II, ingestion of Cd and Zn in feed led to the appearance of highest amounts of granules in the nucleoli, nuclei, and cytoplasm of cells in testes, kidneys, and pancreas. Following Cd intake, the highest accumulation of granules was observed in the nucleoli of hepatocytes and cells of the proximal and distal renal tubules. Combined Cd and Cu produced the highest number of granules in cells of the proximal and distal renal tubules and in the nucleoli and nuclei of germinal epithelium. PMID- 11921346 TI - Distribution of exogenous heavy metals in the hepatocytes of calves: a morphometric study. AB - The objective of this study was to evaluate the presence of heavy metals in the hepatocytes of the animals fed a cadmium-supplemented diet and also receiving zinc and/or selenium in the injection form. The experiment involved four groups of calves (6-8, both sexes) receiving the heavy metals in various combinations for 95 days. Electron micrographs of liver cells were prepared and statistically evaluated using Student's t-test. A modified morphometric apparatus was used for morphometric examination. Exogenous cadmium showed marked accumulation in the hepatocytes. If, however, the cadmium diet was combined with zinc or selenium administration the amount of the reduction product was much lower. PMID- 11921347 TI - Development of a web-based SEM specifically for K-12 education. AB - The scanning electron microscope (SEM) is uniquely suited for use in education due to its ability to produce clear three-dimensional-looking images of virtually any sample. Elementary and secondary science textbooks regularly contain SEM images of bugs, plants, human tissue, rocks, etc. as a means of illustrating the microscopic world to students. However, despite the widespread use of SEM images for educational purposes microscope companies have virtually ignored the area of education in their development and design of SEMs, due presumably to a perceived lack of marketing potential in this area. This article presents the results of a program aimed at developing a Web-based SEM that is specifically designed for educational use by students and teachers in pre-college classrooms. While virtually all companies tout some form of remote control, no concerted effort has been made to fully develop these capabilities for use by the education sector. The microscope under development, termed the WebSEM, seeks to remedy this situation and address this neglected market. Remote control of the WebSEM is possible by means of a simple web interface that allows the users a wide range of controls, depending on the skill of the operator at the remote site. The web interface is specifically designed to be simple and reliable such that little or no training is required for use. This joint effort between educators and a microscope manufacturer is the first attempt at the development of a truly education-oriented SEM suitable for use in the classroom. PMID- 11921348 TI - Application of immunoelectron microscopy techniques in the diagnosis of phytoplasma diseases. AB - An immunoelectron microscopy technique was applied to label Chrysanthemum leuchanthemum phytoplasma in infected leaf tissues of Chrysanthemum leuchanthemum L. and Catharanthus roseus L. plants. Specific monoclonal antibodies at different dilutions and secondary antimouse antibody conjugated with colloidal gold particles of different sizes were used. The monoclonal antibodies demonstrated their specificity against the antigen; immunocytological methods permitted the precise localization and identification of phytoplasmas in thin sections from infected tissues. PMID- 11921349 TI - Adenovirus infection targets the cellular protein kinase CK2 and RNA-activated protein kinase (PKR) into viral inclusions of the cell nucleus. AB - The effects of the adenovirus infection on the distribution of the cellular protein kinase CK2 and double-stranded RNA-activated protein kinase (PKR) were examined at the ultrastructural level. Immunogold labeling revealed the redistribution of CK2 subunits and PKR to morphologically distinct structures of the cell nucleus. The electron-clear amorphous structures, designated pIX nuclear bodies in our previous work (Rosa-Calatrava et al., 2001), contained CK2 alpha and PKR. The protein crystals, which result from the regular assembly of hexon, penton base, and fiber proteins [Boulanger et al. (1970) J Gen Virol 6:329-332], contained CK2 beta and PKR. Both viral structures were devoid of viral RNA, including the PKR-inhibitor VA1 RNA generated by the RNA polymerase III. Instead, VA1 RNA accumulated in PKR-free viral compact rings in which the viral RNA generated by the RNA polymerase II was excluded. PMID- 11921350 TI - Biology of adrenomedullin. Introduction. PMID- 11921351 TI - Adrenomedullin and PAMP: discovery, structures, and cardiovascular functions. AB - We discovered adrenomedullin (AM) from human pheochromocytoma tissue by monitoring the elevating activity of intracellular cyclic AMP (cAMP) in rat platelets in 1993. Since the discovery of AM, it has attracted intense interest from cardiovascular researchers because AM elicits multiple biological activities, including a potent and powerful hypotensive activity caused by dilatation of resistance vessels. AM is biosynthesized and secreted from tissues, including cardiovascular organs. In addition to AM, "proadrenomedullin N-terminal 20 peptide (PAMP)," another biologically active peptide, was found to be processed from the AM precursor. Plasma AM levels are increased in various cardiovascular and renal diseases. AM, therefore, seems to function as a novel system that controls circulation and body fluid, and may be involved in pathophysiological changes in cardiovascular diseases. Therefore, in this review we will focus on the structure of AM and its gene, distribution, receptor, and the physiological and pathological roles of AM in cardiovascular disease. PMID- 11921352 TI - Functional interaction of G protein-coupled receptors of the adrenomedullin peptide family with accessory receptor-activity-modifying proteins (RAMP). AB - Adrenomedullin (AM), alpha- and beta-calcitonin gene-related peptide (CGRP), calcitonin (CT), and amylin are homologous polypeptides with overlapping biological actions such as vasodilatation and inhibition of bone resorption. They are brought about through receptors that include the CT receptor (CTR) and an initially orphan CT receptor-like receptor (CRLR) in association with receptor activity-modifying proteins (RAMP)1, -2, and -3. Co-expression of CRLR with RAMP1 or -2 revealed CGRP or AM receptors, respectively. The CTR interacts with CT and does not require a known RAMP for functional expression. The same CTR is a CGRP/amylin or an amylin receptor upon co-expression with RAMP1 or -3, respectively. Interactions between CRLR and RAMP are thought to be required for their delivery to the cell surface. There, heterodimeric complexes between CRLR or CTR and the corresponding RAMP reveal high-affinity receptors for AM, CGRP, and amylin. Here we review the current knowledge on interactions of G protein coupled receptors with defined associated proteins. PMID- 11921353 TI - Identification, characterization, and physiological actions of factor H as an adrenomedullin binding protein present in human plasma. AB - A recently discovered adrenomedullin binding protein has been characterized as complement factor H, an important regulator of the complement cascade. This review will describe the evidence that led to the identification of factor H as an adrenomedullin binding protein and will address the implications that such binding has in the radioimmunoassay of AM in plasma. We will also describe the possible physiological implications of AM binding: namely, factor H suppresses the antimicrobial activity of AM, enhances AM-mediated induction of cyclic-AMP in rat fibroblasts, and augments the AM-mediated growth of a human cancer cell line. These initial studies suggest that factor H may be an important factor in the regulation of AM physiology. The elucidation of the mechanisms that modulate AM activity will be necessary for the understanding of the role of AM in normal and pathological conditions. PMID- 11921354 TI - Regulation of adrenomedullin expression and release. AB - Adrenomedullin (AM) was originally identified in the extracts of human pheochromocytoma tissue, but this peptide is now known to be synthesized and secreted from many kinds of cells in the body, including vascular smooth muscle cells, endothelial cells, fibroblasts, cardiac myocytes, epithelial cells, and cancer cells. In this review, we summarize AM-secreting and AM gene-expressing cells in addition to the regulation of secretion and gene expression of AM. Although the data are still limited to deduce the general features of AM gene expression, synthesis, and secretion, AM is assumed to be classified into the new class of biologically active peptides, which is mainly expressed and secreted from non-endocrine type cells by the stimulation with inflammation-related substances. It is also interesting that serious physiological conditions such as inflammation or hypoxia potently stimulate AM expression and release, suggesting its unique physiological function distinct from other known biologically active peptides. PMID- 11921355 TI - Adrenomedullin in mammalian embryogenesis. AB - Here are summarized data supporting that adrenomedullin (AM) is a multifunctional factor involved in the complex regulatory mechanisms of mammalian development. During rodent embryogenesis, AM is first expressed in the heart, followed by a broader but also defined spatio-temporal pattern of expression in vascular, neural, and skeletal-forming tissues as well as in the main embryonic internal organs. AM pattern of expression is suggestive of its involvement in the control of embryonic invasion, proliferation, and differentiation processes, probably through autocrine or paracrine modes of action. AM levels in fetoplacental tissues, uterus, maternal and umbilical plasma are highly increased during normal gestation. These findings in addition to other physiological and gene targeting studies support the importance of AM as a vasorelaxant factor implicated in the regulation of maternal vascular adaptation to pregnancy, as well as of fetal and fetoplacental circulations. AM is also present in amniotic fluid and milk, which is suggestive of additional functions in the maturation and immunological protection of the fetus. Altered expression of AM has been found in some gestational pathologies, although it is not yet clear whether this corresponds to causative or compensatory mechanisms. Future studies in regard to the distribution and expression levels of the molecules known to function as AM receptors, together with data on the action of complement factor H (an AM binding protein), may help to better define the roles of AM during embryonic development. PMID- 11921356 TI - Multiple roles of adrenomedullin revealed by animal models. AB - The newly identified adrenomedullin (AM) gene codes for a potent, highly conserved vasodilator that is expressed in many tissues. Many biological functions have been ascribed to AM based on its broad expression pattern and numerous in vitro studies, and it is currently viewed as a multifunctional peptide hormone. Recent advances in gene manipulation have permitted the development of experimental animal systems to help distinguish between gene causes and effects in the context of otherwise normal physiology, and so the normal biological function of the AM gene can be studied within the intact physiological milieu of a whole animal. In this review article, we summarize the recent findings from three different types of genetic experiments involving the AM gene. PMID- 11921357 TI - Adrenomedullin as a pancreatic hormone. AB - Adrenomedullin (AM) is a multiregulatory peptide which is expressed in a wide range of tissues. In the pancreas, AM was first found in mammals, including man, and its colocalization with the pancreatic polypeptide (PP) was established in islet F cells. In addition, three different AM receptors have been characterized in B-cells. AM has been also located in the pancreatic cells of other vertebrate classes. The frequency and distribution of AM cells vary between different animals; they can be found scattered among the exocrine tissue, in the islets, or in ductal epithelia. The colocalization of AM with other hormones presents different patterns, although in birds, as in mammals, it seems to colocalize only with PP. The best-determined pancreatic AM function is the inhibition of insulin secretion in B-cells, which seems to be linked to a recently discovered binding protein, factor H. In relation to this physiological role, clinical data show that AM is raised in some groups of both types I and II diabetic patients and AM might have triggered the disease in a subset of them. On the other hand, AM pancreatic cells are also involved in the response to septic shock by increasing AM circulating levels. A third putative function is the inhibition of amylase secretion by the exocrine pancreatic cells. AM has been found in embryonic mammalian pancreas from the earliest stages of the development, colocalizing with all pancreatic hormones, although in adults only coexpression with PP is kept. AM may play a role in the growth and morphogenesis of the pancreas. PMID- 11921358 TI - Adrenomedullin in the central nervous system. AB - Adrenomedullin (AM) is a novel vasodilator peptide first purified from human pheochromocytoma by tracing its capacity to stimulate cAMP production in platelets. AM immunoreactivity is widely distributed in the central nervous system (CNS) and in the rat has been demonstrated by immunohistochemical techniques to be present in many neurons throughout the brain and spinal cord, as well as in some vascular endothelial cells and perivascular glial cells. Electron microscopy shows that the immunoreactivity is located mainly in the neuronal cytoplasm, but also occurs in the cell nucleus in some cells of the caudate putamen and olfactory tubercle. Biochemical analyses suggest that higher molecular forms, presumably precursor forms, may predominate over fully processed AM in some brain areas. The expression of AM immunoreactivity is increased in cortical neurons, endothelial cells, and perivascular processes after a simulation of ischemia by oxygen and glucose deprivation. Immunohistochemical, electrophysiological, and pharmacological studies suggest that AM in the CNS can act as a neurotransmitter, neuromodulator, or neurohormone, or as a cytoprotective factor in ischemic/hypoxic conditions, in addition to its vasodilator role. PMID- 11921359 TI - Adrenomedullin in the adrenal. AB - Adrenomedullin (AM) was originally characterized in extracts of an adrenal medullary tumor. Since this original finding the peptide and its mRNA have also been found in the adrenal cortex, specifically, in the cells of the aldosterone secreting zona glomerulosa. It is clear that the synthesis of AM is actively regulated in both cortex and medulla. Much research effort has been focused on identifying a role for AM in the adrenal gland. To date, no consistent effect on medullary catecholamine biosynthesis has been demonstrated. In the cortex the actions of AM are controversial and appear to depend on both the tissue preparation used and on the specific receptor population expressed in the individual gland. The results of further studies on the long-term actions of AM on adrenal growth and differentiation are awaited with interest. PMID- 11921360 TI - Adrenomedullin and proadrenomedullin N-terminal 20 peptide in the normal prostate and in prostate carcinoma. AB - There is increasing evidence for the important role played by regulatory peptides in the physiology of the normal and neoplastic prostate. Adrenomedullin (AM) and pro-adrenomedullin N-terminal 20 peptide (PAMP) are recently discovered regulatory peptides widely expressed in the normal prostate and in prostate carcinoma. AM is produced in secretory, stroma, and endothelial cells and in neurons of the prostate ganglia. PAMP is only produced by neuroendocrine cells. The expression of AM mRNA is regulated by androgens in the rat prostate. The number of neuroendocrine cells expressing PAMP is increased in prostate carcinoma after androgen deprivation, which shows that this peptide could regulate androgen independent prostate tumor growth. However, the roles of AM and PAMP in the normal prostate and in prostate carcinoma are yet to be elucidated. PMID- 11921362 TI - Adrenomedullin functions as an important tumor survival factor in human carcinogenesis. AB - Adrenomedullin (AM) is a pluripotent regulatory peptide initially isolated from a human pheochromocytoma (adrenal tumor) and subsequently shown to play a critical role in cancer cell division, tumor neovascularization, and circumvention of programmed cell death, thus it is an important tumor cell survival factor underlying human carcinogenesis. A variety of neural and epithelial cancers have been shown to produce abundant amounts of AM. Recent findings have implicated elevation of serum AM with the onset of malignant expression. In addition, patients with tumors producing high levels of this peptide have a poor prognostic clinical outcome. Given that most human epithelial cancers display a microenvironment of reduced oxygen tension, it is interesting to note that AM and several of its receptors are upregulated during hypoxic insult. The existence of such a regulatory pathway has been implicated as the basis for the overexpression of AM/AM-R in human malignancies, thereby generating a subsequent autocrine/paracrine growth advantage for the tumor cell. Furthermore, AM has been implicated as a potential immune suppressor substance, inhibiting macrophage function and acting as a newly identified negative regulator of the complement cascade, protective properties which may help cancer cells to circumvent immune surveillance. Hence, AM's traditional participation in normal physiology (cited elsewhere in this issue) can be extended to a primary player in human carcinogenesis and may have clinical relevance as a biological target for the intervention of tumor progression. PMID- 11921363 TI - Adrenomedullin has multiple roles in disease stress: development and remission of the inflammatory response. AB - The upregulation of adrenomedullin (AM) gene expression and increases in systemic circulatory as well as localized tissue AM concentrations is well coordinated with the onset and progression of trauma, infection, and sepsis. As such, the coordinated change in AM suggests a key role for this peptide in the inflammatory response. By clinical definition, the process of inflammation constitutes an orchestrated cascade of localized tissue and systemic responses to immunological challenges. Classical responses to the onset of disease stresses are manifested in the timely elaboration of humoral, blood-borne signal effectors (such as adrenocortical and locally produced tissue hormones, immune cytokines, and inorganic signals such as nitric oxide) as well as patterned migration and infiltration of circulating bone marrow-derived cells (mononuclear cells such as monocyte-macrophages and polymorphonuclear cells like neutrophils) largely associated with or delivered through the vascular system. The body's attempts to combat acute infection to restore homeostatic equilibrium are further compromised by underlying disease situations. Atherosclerosis, diabetes, and cardiovascular disease, as well as nutritional metabolic derangements and persistent subclinical infection perturb the regulatory feedback loops necessary for proper control of response effectors like hormones and cytokines. When imbalances occur, tissue necrosis can ensue as driven by free radical damage to cell components. A true appreciation of the inflammatory response can only be grasped through an integrative approach in which the relationship between the different physiological systems is viewed in terms of a changing, dynamic interaction. In essence, the inflammatory response can be thought of in three phases: a period of severity assessment, a period of remediation, and a period of homeostatic restoration. Indeed, AM has differential effects on cellular metabolism, immune function, endocrine function, and cardiovascular function. This peptide appears to play a pivotal role in both reprioritizing the biological needs of tissues and organs during the three phases of inflammatory response as well as a role in restoring homeostatic equilibrium to the body. PMID- 11921361 TI - Adrenomedullin and the integrative physiology of fluid and electrolyte balance. AB - Adrenomedullin (AM) is hypothesized to be a physiologically relevant regulator in fluid and electrolyte homeostasis. AM acts within the central nervous system to inhibit both water and salt intake. The peptide has direct actions in the hypothalamus to decrease vasopressin secretion and in the pituitary gland to inhibit ACTH release. AM decreases production and release of aldosterone from the adrenal glands and acts directly in the kidneys to increase renal blood flow and cause diuresis and natriuresis. Whether or not these complementary actions in brain, pituitary, adrenal gland, and kidney reflect coordinated regulatory mechanisms is currently unknown. Development of molecular tools to determine the physiologic role of endogenous AM will greatly enhance our understanding of AM and its regulation of fluid and electrolyte homeostasis. PMID- 11921364 TI - Interfaces for coupled liquid-phase separation/mass spectrometry techniques. An update on recent developments. AB - An update is presented covering the latest developments in the interfacing of liquid-phase separation systems and mass spectrometers. The interfacing devices presented are those developed for continuous-flow matrix-assisted laser desorption/ionization, micro- and nano-liquid chromatography/masspectrometry (MS), capillary electrophoresis/MS and on-chip separation technologies/MS. From the information that can be found in the most recent literature on the topic, it is evident that the trend towards the miniaturization of separation and interface devices is gaining ground. This can be rationalized by the substantial gains in sensitivity for the detection and study of extremely low levels of analytes and especially of high molecular mass biopolymers. PMID- 11921365 TI - Infrared laser desorption and ionization of polypeptides from a polyacrylamide gel. AB - We observed direct desorption and ionization of angiotensin II and bovine insulin from a frozen polyacrylamide gel without the addition of an exogenous matrix, using picosecond pulses from a tunable, mid-infrared free-electron laser tuned to strong absorption bands of the gel. At 5.7, 5.9, 6.1 and 6.3 microm we were able to desorb and ionize both analyte molecules, with the strongest analyte signal generated at 5.9 microm. However, no analyte signal was observed at 5.5 microm. Consistent with a previous report, we did not observe ions of either polypeptide at 2.9 microm, in spite of strong overall absorption. We discuss the implications of this wavelength-dependent ionization, including possible ablation mechanisms and energy partitioning between competing vibrational modes. PMID- 11921366 TI - Structural characterization of lipopeptide biomarkers isolated from Bacillus globigii. AB - Spectra obtained using matrix-assisted laser desorption/ionization (MALDI) mass spectrometry of Bacillus globigii (Bacillus subtilis niger) spores, vegetative cells and the culture supernatant show a cluster of biomarkers centered at a molecular mass of 1478 Da. Three biomarkers were isolated from the cell-free culture supernatant by solid-phase extraction and reversed-phase high-performance liquid chromatography, and characterized using various kinds of mass spectrometry. A Fourier transform mass spectrometer with a MALDI source was used to determine the monoisotopic protonated masses at 1463.8, 1477.8, and 1505.8 Da in order of elution. The mass differences of 14 and 28 Da suggest that they are homologous molecules. Alkaline hydrolysis of each species showed that it contained a lactone linkage. Strong acid hydrolysis released a fatty acid from an amide bond, consistent with a lipopeptide. A quadrupole time-of-flight instrument with a nanospray source was used to sequence the hydrolyzed forms of the three biomarkers. The cyclic lipopeptides were found to have amino acid sequences identical with those in fengycins and plipastatins, antimicrobial compounds with phospholipase inhibitor activity, previously identified in related species of Bacillus subtilis and Bacillus cereus. PMID- 11921367 TI - Mechanism of azole antifungal activity as determined by liquid chromatographic/mass spectrometric monitoring of ergosterol biosynthesis. AB - A liquid chromatography/mass spectrometry (LC/MS) method for separation and characterization of ergosterol biosynthetic precursors was developed to study the effect of Posaconazole on sterol biosynthesis in fungi. Ergosterol biosynthetic precursors were characterized from their electron ionization mass spectra acquired by a normal-phase chromatography, particle beam LC/MS method. Fragment ions resulting from cleavage across the D-ring and an abundant M - 15 fragment ion were diagnostic for methyl substitution at C-4 and C-14. Comparison of the sterol profile in control and treated Candida albicans incubations showed depletion of ergosterol and accumulation of C-4 and C-14 methyl-substituted sterols following treatment with Posaconazole. These C-4 and C-14 methyl sterols are known to be incapable of sustaining cell growth. The results demonstrate that Posaconazole exerts its antifungal activity by inhibition of ergosterol biosynthesis. Furthermore, Posaconazole appears to disrupt ergosterol biosynthesis by inhibition of lanosterol 14alpha-demethylase. PMID- 11921368 TI - Identification and sequencing analysis of intact proteins via collision-induced dissociation and quadrupole time-of-flight mass spectrometry. AB - Identifying unknown proteins has become a central focal point for proteomic and biopharmaceutical development laboratories. Our laboratory investigated using quadrupole time-of-flight mass spectrometry (Qq/TOFMS) for the analysis of intact proteins for the purpose of identifying unknowns while limiting the number of sample-handling steps between protein extraction and identification. Eight standard proteins, both unmodified and disulfide-bonded and ranging in mass from 5 to 66 kDa, were analyzed using nanoelectrospray and collision-induced dissociation to generate peptide sequence tags. An MS analysis, followed by MS/MS analyses on two to five individual protein charge states, were obtained to make an identification. Peptide sequence tags were extracted from the MS/MS data and used, in conjunction with molecular mass and source origin, to obtain protein identifications using the web-based search engine ProteinInfo (www.proteometrics.com). All of the proteins were unambiguously identified from the input data, after which, all of the major product ions were identified for structural information. In most cases, N- and/or C-terminal ions, and also stretches of consecutive product ions from the protein interior, were observed. This method was applied to the analysis and identification of an unknown detected via reversed-phase high-performance liquid chromatography. PMID- 11921369 TI - Rapid screening and identification of cytochalasins by electrospray tandem mass spectrometry. AB - The cytochalasin class of fungal metabolites was analyzed by electrospray ionization tandem mass spectrometry (ESI-MS/MS) with the aim of developing a methodology for their rapid identification in microbial extracts. ESI-MS analyses of reference cytochalasins were performed and several product ions were produced in MS/MS experiments on parent ions that are structurally characteristic. A precursor ion search was performed to detect cytochalasins in an ethyl acetate extract of fungal strain RK97-F21. Three cytochalasins were detected and one of the components was identified as epoxycytochalasin H by comparing the tandem mass spectra of the product ions with those of reference compounds. This finding was further validated by LC/MS and LC/MS/MS experiments. PMID- 11921370 TI - Role of amino acid N-methylation in cyclosporins on ring opening and fragmentation mechanisms during collisionally induced dissociation in an ion trap. AB - The product ion mass spectra (collisionally induced dissociation mass spectra) of 12 different cyclosporins modified at every N-methylated amino acid residue with respect to cyclosporin A were compared and the effect of N-demethylation on ring opening mechanisms was evaluated. The four preferential protonation sites were identified in [MeBmt(1)]-cyclosporins. Three sites represented the N-methylated nitrogens of Sar(3), MeLeu(6) and MeLeu(9), while the remaining one represented the lactone group formed by the intramolecular N,O-acyl shift. Selective N demethylation resulted either in the deletion of the entire fragment ion series or its substantial attenuation. The structures of three new natural cyclosporins in the study were supported by NMR data. PMID- 11921372 TI - A B-TOF mass spectrometer for the analysis of ions with extreme high start-up energies. AB - Weak magnetic deflection is combined with two acceleration stage time-of-flight mass spectrometry and subsequent position-sensitive ion detection. The experimental method, called B-TOF mass spectrometry, is described with respect to its theoretical background and some experimental results. It is demonstrated that the technique has distinct advantages over other approaches, with special respect to the identification and analysis of very highly energetic ions with an initially large energy broadening (up to 1 MeV) and with high charge states (up to 30+). Similar energetic targets are a common case in intense laser-matter interaction processes found during laser ablation, laser-cluster and laser molecule interaction and fast particle and x-ray generation from laser-heated plasma. PMID- 11921371 TI - Metastable ion study of organosilicon compounds. Part XIV-trimethylsilylacetic acid, (CH(3))(3)SiCH(2)COOH, and its methyl ester, (CH(3))(3)SiCH(2)COOCH(3). AB - The unimolecular metastable decompositions of trimethylsilylacetic acid, (CH(3))(3)SiCH(2)COOH (1), and its methyl ester, (CH(3))(3)SiCH(2)COOCH(3) (2), were investigated by mass-analyzed ion kinetic energy (MIKE) spectrometry in conjunction with thermochemical data. The abundance of the molecular ions of both compounds, generated by electron ionization, is extremely low. However, the abundance of the ions generated by the loss of (.)CH(3) and observed at m/z 117 and 131 is moderate. These fragment ions further decompose to form the most abundant m/z 75 and 89 ions, respectively, by the loss of CH(2)CO through a (CH(3))(2)Si group migration. The loss of CH(2)CO is also observed to occur from 2(+.) and its fragment ion at m/z 115 generated by the loss of (.)OCH(3). The former reaction is proposed to occur via an ion-radical complex. PMID- 11921373 TI - Resonant electron capture mass spectrometry of free fatty acids: examination of ion structures using deuterium-labeled fatty acids and collisional activation. AB - The structures of [M-H](-) ions generated from free fatty acids in resonant electron capture at energies of 1.2 and 7.2 eV were investigated using deuterium labeled isotopomers and collision-induced dissociation. The [M-H](- small middle dot) ions occur in both a carboxylate anion and a carbanion form. While the formation of the carboxylate anion at 1.2 eV involves the loss of a carboxylic hydrogen, that at 7.2 eV involves the loss of a hydrogen from different positions in the aliphatic chain followed by a rearrangement of a carboxylic hydrogen on to the radical site in the chain. The [M-H-H(2)O](-) ion which is a minor ion in the resonant electron capture spectrum at 7.2 eV is shown to be a precursor for the charge-remote fragment ions corresponding to formal losses of a hydrogen and elements of alkanes. The [M-H-H(2)O](-) ion corresponding to the second major ion in the resonant electron capture spectrum at 7.2 eV is demonstrated to be consistent with a cyclopentanone anion structure. On the basis of new insights obtained in the present study and taking into account previous results, an updated proposal is presented for the mechanism of charge-remote fragmentation which operates in resonant electron capture of free fatty acids at 7.2 eV. PMID- 11921375 TI - In-gel deglycosylation of sodiumdodecyl sulfate polyacrylamide gel electrophoresis-separated glycoproteins for carbohydrate estimation by matrix assisted laser desorption/ionization time-of-flight mass spectrometry. AB - Mass determination by mass spectrometric methods (electrospray ionization mass spectrometry (ESI-MS), matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOFMS)) of sodiumdodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE)-separated proteins is a well known procedure and reliable protocols are available. In our efforts to use the established methods to determine the molecular mass of the disulfide bridged, heterodimeric glycoprotein GP3 and to determine the carbohydrate content of each protein subunit we developed an in-gel chemical deglycosylation method. For this purpose we established experimental conditions that allow maximum extraction of the high molecular mass protein subunits and developed a routine method to apply the HF pyridine deglycosylation protocol to proteins isolated from polyacrylamide gel pieces. The novel protocol and extraction procedure described can be used to analyze O-glycosylated proteins up to 150 kDa after SDS-PAGE separation. PMID- 11921376 TI - Substituent effect and multisite protonation in the fragmentation of alkyl benzoates. AB - The dissociation of protonated alkyl benzoates (para H, CN, OMe and NO(2)) into protonated benzoic acids and alkyl cations was studied in the gas phase. It was found that the product ratio depends on the substituent at the para position of the phenyl ring. The substituent effect is probably the result of the formation of an ion-neutral complex intermediate that decomposes to an ion and a neutral, according to the relative proton affinities of the two moieties. The experimental results and theoretical calculations indicate that the favored protonation site in these compounds is the ester's carbonyl and that proton transfer from the phenyl ring to the ester group is very likely to occur under chemical ionization conditions. It is most probable that the carbonyl protonated form is a common intermediate in the fragmentation process, regardless of the protonation site. PMID- 11921374 TI - Improved in-gel approaches to generate peptide maps of integral membrane proteins with matrix-assisted laser desorption/ionization time-of-flight mass spectrometry. AB - This paper reports studies of in-gel digestion procedures to generate MALDI-MS peptide maps of integral membrane proteins. The methods were developed for the membrane domain of the mannitol permease of E. coli. In-gel digestion of this domain with trypsin, followed by extraction of the peptides from the gel, yields only 44% sequence coverage. Since lysines and arginines are seldomly found in the membrane-spanning regions, complete tryptic cleavage will generate large hydrophobic fragments, many of which are poorly soluble and most likely contribute to the low sequence coverage. Addition of the detergent octyl-beta glucopyranoside (OBG), at 0.1% concentration, to the extraction solvent increases the total number of peptides detected to at least 85% of the total protein sequence. OBG facilitates the recovery of hydrophobic peptides when they are SpeedVac dried during the extraction procedure. Many of the newly recovered peptides are partial cleavage products. This seems to be advantageous since it generates hydrophobic fragments with a hydrophilic solubilizing part. In-gel CNBr cleavage resulted in 5-10-fold more intense spectra, 83% sequence coverage, fully cleaved fragments and no effect of OBG. In contrast to tryptic cleavage sites, the CNBr cleavage sites are found in transmembrane segments; cleavage at these sites generates smaller hydrophobic fragments, which are more soluble and do not need OBG. With the results of both cleavages, a complete sequence coverage of the membrane domain of the mannitol permease of E. coli is obtained without the necessity of using HPLC separation. The protocols were applied to two other integral membrane proteins, which confirmed the general applicability of CNBr cleavage and the observed effects of OBG in peptide recovery after tryptic digestion. PMID- 11921378 TI - Current awareness. PMID- 11921379 TI - Overview: infection during pregnancy and neurologic outcome in the child. PMID- 11921380 TI - Intrauterine infection and prematurity. AB - Intrauterine infection is a major cause of premature labor with and without intact membranes. Intrauterine infection is present in approximately 25% of all preterm births and the earlier the gestational age at delivery, the higher the frequency of intra-amniotic infection. Microorganisms may also gain access to the fetus before delivery. A fetal inflammatory response syndrome elicited in response to microbial products is associated with the impending onset of preterm labor and also with multi-systemic organ involvement in the human fetus and a higher rate of perinatal morbidity. The most common microorganisms involved in intrauterine infections are Ureaplasma urealyticum, Fusobacterium species and Mycoplasma hominis. The role of Chlamydia trachomatis and viruses in preterm labor remain to be determined. Use of molecular microbiology techniques to diagnose intrauterine infection may uncover the role of fastidious microorganisms that have not yet been discovered. Antibiotic administration to patients with asymptomatic bacteriuria is associated with a significant reduction in the rate of preterm birth. However, such benefit has not been demonstrated for patients with bacterial vaginosis, or women who carry Streptococcus agalactia, Ureaplasma urealyticum or Trichomonas vaginalis. Antibiotic administration to patients with preterm premature rupture of membranes is associated with prolongation of pregnancy and a reduction in the rate of clinical chorioamnionitis and neonatal sepsis. The benefit has not been demonstrated in patients with preterm labor and intact membranes. Major efforts are required to determine why some women develop an ascending intrauterine infection and others do not and also what interventions may reduce the deleterious effect of systemic fetal inflammation. PMID- 11921381 TI - Infection markers and early signs of neonatal encephalopathy in the term infant. AB - Recent evidence points to an association between intrauterine infection and cerebral palsy (CP) in the preterm as well as the term infant. The mechanisms that link these two conditions are unclear. Chorioamnionitis is a common clinical problem complicating 5-10% of pregnancies, whereas the incidence of CP attributed to intrapartum asphyxia is rare. Chorioamnionitis may result in brain injury as a result of interruption of placental blood flow (asphyxia), or via fever and/ or the cytokine release associated with infection. This review will attempt to establish the link between perinatal infection and brain damage in term infants. The characteristics of the perinatal inflammatory response, the potential mechanisms of brain injury associated with infection, and the clinical characteristics of neonatal encephalopathy will be discussed. PMID- 11921382 TI - Neonatal encephalopathy in the term infant: neuroimaging and inflammatory cytokines. AB - The interrelationship between inflammation and ischemia is complex and poorly understood in the developing nervous system. In the preterm newborn, maternal infection may predispose to white matter injury and may be associated with cytokine elevation. In the term infant, few studies exist linking elevation of cytokines with encephalopathy and poor neurodevelopmental outcome. This review discusses the interplay among inflammatory cytokines, neonatal encephalopathy, and neuroimaging parameters. PMID- 11921383 TI - Systematic review of chorioamnionitis and cerebral palsy. AB - In a recent meta-analysis evaluating the relationship between chorioamnionitis and cerebral palsy, we found that chorioamnionitis is a risk factor for both cerebral palsy and cystic periventricular leukomalacia (cPVL). The current paper extends the meta-analysis by including studies published in the year 2000, and by further evaluating the causes of heterogeneity among individual study results. Using a random effects model, clinical chorioamnionitis was significantly associated with both cerebral palsy (RR 1.9, 95% CI 1.5-2.5) and cPVL (RR 2.6, 95% CI 1.7-3.9). Sources of heterogeneity included widely varying practices in the diagnosis of clinical chorioamnionitis, different gestational age characteristics, and varying study year. We conclude that based on the available literature, chorioamnionitis is a risk factor for both cerebral palsy and cPVL. PMID- 11921384 TI - Models of white matter injury: comparison of infectious, hypoxic-ischemic, and excitotoxic insults. AB - White matter damage (WMD) in preterm neonates is strongly associated with adverse outcome. The etiology of white matter injury is not known but clinical data suggest that ischemia-reperfusion and/or infection-inflammation are important factors. Furthermore, antenatal infection seems to be an important risk factor for brain injury in term infants. In order to explore the pathophysiological mechanisms of WMD and to better understand how infectious agents may affect the vulnerability of the immature brain to injury, numerous novel animal models have been developed over the past decade. WMD can be induced by antenatal or postnatal administration of microbes (E. coli or Gardnerella vaginalis), virus (border disease virus) or bacterial products (lipopolysaccharide, LPS). Alternatively, various hypoperfusion paradigms or administration of excitatory amino acid receptor agonists (excitotoxicity models) can be used. Irrespective of which insult is utilized, the maturational age of the CNS and choice of species seem critical. Generally, lesions with similarity to human WMD, with respect to distribution and morphological characteristics, are easier to induce in gyrencephalic species (rabbits, dogs, cats and sheep) than in rodents. Recently, however, models have been developed in rats (PND 1-7), using either bilateral carotid occlusion or combined hypoxia-ischemia, that produce predominantly white matter lesions. LPS is the infectious agent most often used to produce WMD in immature dogs, cats, or fetal sheep. The mechanism whereby LPS induces brain injury is not completely understood but involves activation of toll-like receptor 4 on immune cells with initiation of a generalized inflammatory response resulting in systemic hypoglycemia, perturbation of coagulation, cerebral hypoperfusion, and activation of inflammatory cells in the CNS. LPS and umbilical cord occlusion both produce WMD with quite similar distribution in 65% gestational sheep. The morphological appearance is different, however, with a more pronounced infiltration of inflammatory cells into the brain and focal microglia/macrophage ("inflammatory WMD") in response to LPS compared to hypoperfusion evoking a more diffuse microglial response usually devoid of cellular infiltrates ("ischemic WMD"). Furthermore, low doses of LPS that by themselves have no adverse effects in 7-day-old rats (maturation corresponding to the near term human fetus), dramatically increase brain injury to a subsequent hypoxic-ischemic challenge, implicating that bacterial products can sensitize the immature CNS. Contrary to this finding, other bacterial agents like lipoteichoic acid were recently shown to induce tolerance of the immature brain suggesting that the innate immune system may respond differently to various ligands, which needs to be further explored. PMID- 11921385 TI - Public health issues related to infection in pregnancy and cerebral palsy. AB - Cerebral palsy is the most common neuromotor developmental disability of childhood, affecting as many as 8,000 to 12,000 children born in the U.S. each year (corresponding to a prevalence rate of between 2 and 3 per 1000 children). Recent improvements in neonatal care have not resulted in a decline in the overall prevalence of cerebral palsy and, in fact, greater numbers of very preterm/very low birth weight infants are surviving with cerebral palsy and other developmental problems. Infection in pregnancy may be an important cause of the disorder. In preterm infants, there appears to be about a 2-fold increased risk for cerebral palsy from chorioamnionitis, and in term infants the estimated increased risk is about 4-fold. Provisionally, chorioamnionitis might account for 12% of spastic cerebral palsy in term infants and 28% of cerebral palsy in preterm infants. Studies of biochemical markers of fetal inflammation typically associated with infection also suggest that an inflammatory response may be an important independent etiologic factor. If a substantial proportion of cerebral palsy is attributable to acute amnionitis infection and/or neonatal sepsis, cerebral palsy should have decreased in the United States after administration of intrapartum antibiotics became widespread in response to publication of public health consensus guidelines for Group B streptococcus in 1996. However, failure to detect declines could have a number of explanations and these explanations illustrate the many public health challenges related to intrauterine infection and cerebral palsy. Given the gaps in our current knowledge about intrauterine infection and cerebral palsy, public health recommendations for timely and specific prevention activities are limited at this time. PMID- 11921386 TI - Perinatal infection, fetal inflammatory response, white matter damage, and cognitive limitations in children born preterm. AB - Only sparse information is available about a possible association between antenatal infection outside the brain and subsequent cognitive limitations among preterm infants. Based on published studies, we provide a theoretical schema that links them via the fetal inflammatory response and neonatal white matter damage. We conclude that the relationship between antenatal infection and cognitive limitations deserves much further attention by researchers interested in the prevention of this undesirable outcome of prematurity. PMID- 11921387 TI - In utero infection and adult schizophrenia. AB - We review emerging evidence indicating that in utero exposure to infection is a risk factor for schizophrenia. It is hypothesized that a prenatal infection increases the liability to schizophrenia in adulthood by adversely affecting the maturation of critical brain structural and functional components implicated in the pathogenesis and pathophysiology of the disorder. Early evidence for a role of in utero infection includes investigations linking schizophrenia with birth during the winter and in urban regions, and ecologic studies demonstrating associations between influenza epidemics and births of pre-schizophrenic patients. The findings of the latter studies are, however, equivocal. To more rigorously address this question, our group has used increasingly sophisticated research designs that incorporate more refined measures of exposure and outcome, and continuous follow-up of treated cases. This work has already yielded several intriguing findings, including associations between schizophrenia and two in utero infections--rubella and respiratory infection. We also describe our ongoing birth cohort investigations that are expected to advance this work further, including studies that utilize maternal serum samples drawn during pregnancy of offspring who were later diagnosed with schizophrenia. PMID- 11921390 TI - Combinatorial biosynthesis of antibiotics: challenges and opportunities. AB - Natural products with antibiotic activity have been central agents in human therapeutics over the past fifty years. They are likely to remain crucial in the decades to come. These molecules, often termed secondary metabolites because they are the end products of dedicated metabolic pathways that are turned on when microbes are stressed by environmental factors such as starvation, can achieve considerable architectural and functional group complexity that allows specific targeting. The programmed manipulation of the genes that encode the enzymes in the biosynthetic pathways offers promise for redesign of antibiotic structures to create new activities and overcome bacterial resistance to existing antibiotics. PMID- 11921391 TI - Specific interactions between sense and complementary peptides: the basis for the proteomic code. AB - The discovery of the genetic code was one of the milestone events in biology: a conserved, universal code defining the primary amino acid sequences of all proteins of all organisms. However, this code has been thought to be limited, unable to provide additional information appropriate to defining the three dimensional structure and function of these proteins. This raises important questions. Can there be more to the genetic code? Is there a code embedded within the code? Does a two-dimensional genetic code exist? In our view, the answer to all three of these questions is a qualified "yes". This review describes how sense and complementary peptides coded for by mutually complementary nucleic acid sequences are capable of interacting specifically, thereby suggesting the existence of a second, two-dimensional genetic code (proteomic code). Theories attempting to explain such specific interactions between sense and complementary peptides are discussed including the Mekler-Idlis (M-I) pair theory that suggests that each codon-directed amino acid residue in a sense peptide may make a specific pair-wise interaction with the corresponding complementary codon directed residue in the complementary peptide. In effect, through-space interactions between pairs of amino acid residues are suggested as being specified by the genetic code and its complement. The biological implications of sense/complementary peptide interactions are potentially vast but still to be fully understood and appreciated. That such peptide/peptide interactions could provide the basis for understanding and constructing the proteomic code remains to be properly established but research to date suggests that we should be able to make a start in that direction. PMID- 11921392 TI - Hydrogenases: hydrogen-activating enzymes. PMID- 11921393 TI - The first view of an ABC transporter: the X-ray crystal structure of MsbA from E. coli. PMID- 11921394 TI - Spin labels as a tool to identify and characterize protein-ligand interactions by NMR spectroscopy. AB - NMR spectroscopy based discovery and optimization of lead compounds for a given molecular target requires the development of methods with maximum sensitivity and robustness. It is shown here that organic nitroxide radicals ("spin labels") can be used to boost the sensitivity of NMR spectroscopic screening in drug discovery research. The concept of utilizing spin labels in NMR spectroscopy is summarized, examples for successful first-site and second-site NMR spectroscopic screens are given, and guidelines for linker design are presented. PMID- 11921395 TI - Functional mimicry of a discontinuous antigenic site by a designed synthetic peptide. AB - Functional reproduction of the discontinuous antigenic site D of foot-and-mouth disease virus (FMDV) has been achieved by means of synthetic peptide constructions that integrate each of the three protein loops that define the antigenic site into a single molecule. The site D mimics were designed on the basis of the X-ray structure of FMDV type C-S8c1 with the aid of molecular dynamics, so that the five residues assumed to be involved in antigenic recognition are located on the same face of the molecule, exposed to solvent and defining a set of native-like distances and angles. The designed site D mimics are disulfide-linked heterodimers that consist of a larger unit containing VP2(71 84), followed by a polyproline module and by VP3(52-62), and a smaller unit corresponding to VP1(188-194) (VP=viral protein). Guinea pig antisera to the peptides recognized the viral particle and competed with site D-specific monoclonal antibodies, while inoculation with a simple (not covalently joined to one another) admixture of the three VP1-VP3 sequences yielded no detectable virus specific serum conversion. Similar results have been reproduced in two bovines. Antisera to the peptides also moderately neutralize FMDV in cell cultures and partially protect guinea pigs against challenge with the virus. These results demonstrate functional mimicry of the discontinuous site D by the peptides, which are therefore obvious candidates for a multicomponent, peptide-based vaccine against FMDV. PMID- 11921396 TI - Low micromolar inhibitors of galectin-3 based on 3'-derivatization of N acetyllactosamine. AB - A strategy for generating potential galectin inhibitors was devised based on derivatization at the C-3' atom in 3'-amino-N-acetyllactosamine by using structural knowledge of the galectin carbohydrate recognition site. A collection of 12 compounds was prepared by N-acylations or N-sulfonylations. Hydrophobic tagging of the O-3 atom in the N-acetylglucosamine residue with a stearic ester allowed rapid and simple product purification. The compounds were screened in a galectin-3 binding assay and three compounds with significantly higher inhibitory activities compared to the parent N-acetyllactosaminide were found. These three best inhibitors all carried an aromatic amide at the C-3' position of the galactose moiety, which indicates that favorable interactions were formed between the aromatic group and galectin-3. The best inhibitor had an IC50 value (4.4 microM) about 50 times better than the parent N-acetyllactosaminide, which implies that it has potential as a valuable tool for studying galectin-3 biological functions and also as a lead compound for the development of galectin 3-blocking pharmaceuticals. PMID- 11921397 TI - Quantification of the Raf-C1 interaction with solid-supported bilayers. AB - By use of the quartz crystal microbalance technique, the interaction of the Raf Ras binding domain (RafRBD) and the cysteine-rich domain Raf-C1 with lipids was quantified by using solid-supported bilayers immobilized on gold electrodes deposited on 5 MHz quartz plates. Solid-supported lipid bilayers were composed of an initial octanethiol monolayer chemisorbed on gold and a physisorbed phospholipid monolayer varying in its lipid composition as the outermost layer. The integrity of bilayer preparation was monitored by impedance spectroscopy. For binding experiments, a protein construct comprising the RafRBD and Raf-C1 linked to the maltose binding protein and a His tag, termed MBP-Raf-C1, was used. Dissociation constants and rate constants of the association and dissociation were obtained for various 1,2-dimyristoyl-sn-glycero-3-phosphocholine (DMPC)/1,2 dimyristoyl-sn-glycero-3-phosphoserine (DMPS) lipid mixtures. Independently of the phosphatidylserine (PS) content, the dissociation constants were in the order of 5x10(-7) M, while the on-rate constants were in the range of 2x10(3) (M s)(-1) and the off-rate constants in the range of 1x10(-3) s(-1). The maximum frequency shift increased significantly with increasing amounts of DMPS; this indicates that this negatively charged lipid is the primary binding site for MBP-Raf-C1. Exchange of DMPS for 1,2-dimyristoyl-sn-glycero-3-phosphoglycerol (DMPG) did not alter the thermodynamics and kinetics of protein binding, which implies that the protein interaction is mainly electrostatically driven. Scanning force microscopy (SFM) was employed to render protein adsorption visible and to confirm the assumption of a protein monolayer on the lipid layer. SFM images clearly revealed that the protein binds preferentially, but not solely, to negatively charged phosphatidylserine headgroups. We hypothesize that PS-enriched domains are initial binding sites with high affinity for Raf-C1, but that lateral interactions may account for protein domain growth. PMID- 11921398 TI - Evolution of nitrate reductase: molecular and structural variations on a common function. AB - The biological transformation of nitrogen oxyanions is widespread in nature and gives rise to a robust biogeochemical cycle. The first step in nitrate reduction is carried out by the enzyme nitrate reductase (NR). Although NR always catalyzes the same chemical reaction (conversion of nitrate into nitrite), its location in the cell, structure, and function are organism-dependent. We use protein sequence data to determine phylogenetic relationships and to examine similarities in structure and function. Three distinct clades of NR are apparent: the eukaryotic assimilatory NR (Euk-NR) clade, the membrane-associated prokaryotic NR (Nar) clade, and a clade that includes both the periplasmic NR (Nap) and prokaryotic assimilatory NR (Nas). The high degree of sequence similarity and a phylogenetic distribution that follows taxonomic classification suggest a monophyletic origin for the Euk-NR early on in the evolution of eukaryotic cells. In contrast, sequence conservation, phylogenetic analysis, and physiology suggest that both Nar and Nap were acquired by horizontal gene transfer. Nap and Nas share a lesser degree of similarity, with Nap a subclade of Nas. Nap from strict anaerobic bacteria such as Desulfovibrio desulfuricans is ancestral to facultative species and may provide an evolutionary link between Nap and Nas. We observed conserved binding sites for molybdenum and pterin cofactors in all four proteins. In pathways involving Euk-NR, Nas, and Nar, for which ammonia is the end product, nitrite is reduced to ammonia by a siroheme nitrite reductase. Nap, however, is coupled to a pentaheme nitrite reductase. In denitrification, whether Nar or Nap is involved, nitrite is reduced to nitric oxide by either a cytochrome cd1 or a copper-containing nitrite reductase. This complexity underscores the importance of nitrate reduction as a key biological process. PMID- 11921399 TI - A novel near-infrared fluorescence sensor for detection of thrombin activation in blood. AB - Thrombosis is an important pathophysiologic component of many cardiovascular diseases. Thrombin, a serine protease, plays a central role in thrombosis formation. Detection and imaging of thrombin activity may thus be of considerable biomedical interest. The goal of this study was to design, synthesize, and characterize a novel thrombin-activated near-infrared fluorescence (NIRF) probe. The probe consisted of a thrombin-cleavable peptide spacer (...D-Phe-Pip-Arg...; Pip=pipecolic acid) and contained a terminal fluorescence reporter which was quenched when conjugated to a biocompatible delivery vehicle. A control peptide spacer was synthesized that differed by one amino acid. Following thrombin addition, the probe was activated within minutes. The NIRF signal increased by a factor of 27-fold within 20 minutes, and was inhibited by hirudin, a specific thrombin inhibitor. NIRF optical imaging experiments confirmed rapid activation of the probe in both buffer and human blood. The control probe showed minimal activation in all experiments. In addition to potentially furthering our understanding of thrombin regulation in vivo, the thrombin-activated near infrared probe may have broad clinical application to the diagnosis of arterial and venous thrombosis. PMID- 11921400 TI - Natural deuterium distribution in branched-chain medium-length fatty acids is nonstatistical: a site-specific study by quantitative 2H NMR spectroscopy of the fatty acids of capsaicinoids. AB - Quantitative 2H NMR spectroscopy has been used to determine the natural abundance site-specific 2H isotopic content of 6,7-dihydrocapsaicin (1) and capsaicin (2). Prior to analysis, the fatty acyl moieties were released as methyl 8 methylnonanoate (3) and methyl E-8-methylnon-6-enoate (4), respectively. A marked and similar nonstatistical isotopic distribution of (2)H is observed for both fatty acids. Notably, it can be seen that: 1) the isobutyl portion of 3 is more impoverished in 2H than the methylenic portion; 2) the isobutyl portion of 4 is more impoverished than that of 3; 3) an alternating pattern occurs in the (2H/1H)i between the C3 to C7 positions; and 4) the ethylenic hydrogens at C6 and C7 of 4 are, respectively, impoverished and unchanged relative to these positions in 3. These observations are compatible with the proposed biosynthetic origins of the different parts of 1 and 2, and with the view that 1 is a proximal precursor of 2. Furthermore, it can be suggested that, firstly, the hydrogen atoms at C3 to C7 originate alternatively from the substrate and from the environment and, secondly, that the Delta6-E desaturation is introduced by a mechanism closely mimicking that of the Z desaturation of higher plants. PMID- 11921401 TI - Novel carboranes with a DNA binding unit for the treatment of cancer by boron neutron capture therapy. AB - The synthesis and biological evaluation of two ortho-carborane derivatives which contain a 5,6,7-trimethoxyindole (TMI) unit for use in boron neutron capture therapy is described. The TMI moiety is known to be the DNA-binding part of the highly potent anticancer agent duocarmycin A. The ortho-carborane derivatives were prepared from amino alkynes which were bound to a protected TMI carboxylic acid. Addition of decaborane(14) to the alkyne triple bond with subsequent removal of the tert-butoxycarbonyl (Boc) and benzyl protecting groups gave the desired product. Boron uptake from the ortho-carborane derivatives into B-16 melanoma cells was higher and faster than that observed with L-p boronophenylalanine (BPA), which is in use in the clinic. PMID- 11921402 TI - Characterization and peroxidase activity of a myoglobin mutant containing a distal arginine. AB - The spectroscopic, conformational, and reactivity characteristics of the T67R variant of sperm whale myoglobin have been studied to assess the effects of introducing an arginine residue into the distal side of this protein, as occurs in the active site of heme peroxidases. The overall circular dichroism (CD) and NMR spectroscopic properties of various derivatives of the protein are little affected by the mutation. The mutant contains a high-spin ferric ion with a water molecule as the sixth ligand, which exhibits slightly enhanced acidity (pK(a)=8.43+/-0.03) with respect to the corresponding derivative of wild-type myoglobin (pK(a)=8.60+/-0.04). The presence of the distal arginine increases the affinity of the Fe(III) center for azide (K=(6.0+/-0.5)x10(4) M(-1)) and decreases that for imidazole (K=12.0+/-0.2 M(-1)), with respect to the wild-type protein (K=(5.0+/-0.1)x10(4) and 24.7+/-0.7 M(-1), respectively). The peroxidase activity of T67R and wild-type myoglobins has been studied with a group of phenolic substrates related to tyrosine. The mutant exhibits an increased rate of reaction with hydrogen peroxide (k=1550+/-10 versus 760+/-10 M(-1) x s(-1)) and a generally increased peroxidase activity with respect to wild-type myoglobin. Relaxation measurements of proton nuclei of the phenolic substrates in the presence of either the T67R variant or the wild-type protein show that binding of these molecules occurs at distances of 8-10 A from the iron center, that is, close to the heme pocket, except for p-cresol, which can approach the heme more closely and, therefore, probably enter into the distal cavity. PMID- 11921403 TI - Biosynthesis of proteins incorporating a versatile set of phenylalanine analogues. PMID- 11921404 TI - Replacement of the intervening amino acid sequence of a Syk-binding diphosphopeptide by a nonpeptide spacer with preservation of high affinity. PMID- 11921405 TI - DNA-directed assembly of bienzymic complexes from in vivo biotinylated NAD(P)H:FMN oxidoreductase and luciferase. PMID- 11921406 TI - pH-dependent binding modes observed in trypsin crystals: lessons for structure based drug design. PMID- 11921407 TI - De novo design, synthesis, and in vitro evaluation of inhibitors for prokaryotic tRNA-guanine transglycosylase: a dramatic sulfur effect on binding affinity. PMID- 11921408 TI - Synthesis and characterisation of acyl carrier protein bound polyketide analogues. PMID- 11921409 TI - Cellular uptake studies with beta-peptides. PMID- 11921412 TI - Islet autoantibodies in cord blood: maternal, fetal, or neither? AB - In high-risk type 1 diabetes populations, up to 3% of the general population newborns may express islet autoantibodies in cord blood and the vast majority of those appear to be maternal autoantibodies that disappear usually before the age of 9 months. Despite recent progress in standardization of autoantibody assays, some of the findings appear to be artifacts or non-IgG-mediated binding phenomena. It remains unclear whether transplacentally transmitted maternal autoantibodies play any role in protecting the offspring of diabetic women from diabetes. The evidence for fetal production of islet autoantibodies is very limited and remains to be validated in large prospective studies currently underway. PMID- 11921413 TI - The role of increased liver triglyceride content: a culprit of diabetic hyperglycaemia? AB - The role of fat in the aetiology of insulin resistance and type 2 diabetes has been re-considered in the present review. This is because of the questions raised by recent created mouse models imitating human lipodystrophy diabetes. It appears that hepatic steatosis, which is shared by both lipodystrophy and most if not all obesity patients, may play a key role in the pathogenesis of insulin resistance and type 2 diabetes despite the fact that lipodystrophy is an extreme state and occurs more rarely than obesity. The possible link between lipid and glucose metabolisms via peroxisome activity has been examined and its role in determining hyperglycaemia is suggested. Moreover, new avenues towards a better understanding of insulin resistance at the genomic level have also been proposed. It appears that one of the most fundamental biological phenomena, fuel selection, may underlie the causes of diabetic hyperglycaemia and perplex the role of fat in the aetiology of insulin resistance. PMID- 11921414 TI - The insulin gene in diabetes. AB - Lack of insulin production or abnormalities affecting insulin secretion are key to the development of almost all forms of diabetes, including the common type 1 (insulin-dependent) and type 2 (non-insulin-dependent) diabetes and the more rare forms of maturity-onset diabetes of the young (MODY). Because insulin has such a central role in the pathogenesis of both forms of diabetes, the insulin gene (INS) has always been considered a candidate susceptibility gene. A number of studies have shown that the allelic variation and parent-of-origin effects affect the transmission and expression of the insulin gene in pancreatic beta-cells and extra-pancreatic tissues. These observations have led to the formulation of new hypotheses to explain the biological mechanisms by which functional differences in the expression of the insulin gene may contribute to diabetes susceptibility. PMID- 11921415 TI - Reaching those most in need: a review of diabetes self-management interventions in disadvantaged populations. AB - There has been increased recognition of the importance of developing diabetes self-management education (DSME) interventions that are effective with under served and minority populations. Despite several recent studies in this area, there is to our knowledge no systematic review or synthesis of what has been learned from this research. An electronic literature search identified five formative evaluations and ten controlled DSME intervention trials focused on under-served (low-income, minority or aged) populations. The RE-AIM (Reach, Efficacy, Adoption, Implementation, Maintenance) evaluation framework was used to evaluate the controlled studies on the dimensions of reach, efficacy, adoption, implementation, and maintenance. Fifty percent of the studies identified reported on the percentage of patients who participated, and the percentages were highly variable. The methodological quality of the articles was generally good and the short-term results were encouraging, especially on behavioral outcomes. Data on adoption (representativeness of settings and clinicians who participate) and implementation were almost never reported. Studies of modalities in addition to group meetings are needed to increase the reach of DSME with under-served populations. The promising formative evaluation work that has been conducted needs to be extended for more systematic study of the process of intervention implementation and adaptation with special populations. Studies that explicitly address the community context and that address multiple issues related to public health impact of DSME interventions are recommended to enhance long-term results. PMID- 11921416 TI - Risk assessment for developing type 1 diabetes: intentions of behavioural changes prior to risk notification. AB - BACKGROUND: Recent progress in predictive techniques allows people at risk of developing type 1 diabetes to be identified in a pre-symptomatic stage and prevention trials to be implemented. The present study examined prospectively whether participants in a screening programme anticipated behavioural changes in the event of having a high risk. METHODS: Four hundred and three first-degree relatives of people with type 1 diabetes completed a self-administered questionnaire about their views on screening and diabetes, and questionnaires on well-being and locus of control. RESULTS: Prior to risk notification, 73% reported that they intended to introduce lifestyle changes if at high risk. The vast majority of the respondents (87%) reported that eating habits would be the main changes made. Those anticipating changes believed they could take actions to reduce their risk of type 1 diabetes (p<0.001) and to have personal control over diabetes onset (p<0.001). They were also more worried about developing diabetes (p<0.01) and preoccupied with diabetes-related symptoms (p<0.01). CONCLUSIONS: Prior to risk notification, the process of being screened raised concerns and expectations about future changes. Despite the lack of any evidence, people believed lifestyle changes would be effective in reducing their risk. Since the impact of lifestyle in the development of type 1 diabetes is not yet established, accurate information about the role of health behaviour in the progression to overt diabetes is needed to avoid unrealistic expectations on the benefit of these changes and unnecessary impairment to quality of life. Personally initiated changes should be monitored since they could importantly influence the progress and outcome of prevention trials. PMID- 11921417 TI - Autoantibodies to GAD, IA-2 and insulin in ICA-positive first-degree relatives of children with type 1 diabetes: a comparison between parents and siblings. AB - BACKGROUND: Islet cell antibodies (ICA) represent a heterogenous group of autoantibodies to diabetes-associated antigens, including glutamic acid decarboxylase (GAD) and the IA-2 protein. The objectives of the present study were to compare the prevalence of autoantibodies to known biochemically characterized autoantigens between ICA-positive non-diabetic parents and siblings of children with type 1 diabetes and to evaluate how such antibodies explain ICA reactivity. METHODS: The presence and levels of GAD antibodies (GADA), IA-2 antibodies (IA-2A) and insulin autoantibodies (IAA) were analyzed in the sera of 184 ICA-positive first-degree relatives (79 parents and 105 siblings). RESULTS: The prevalences of GADA (61.9% in siblings vs 32.9% in parents), IA-2A (55.2% vs 15.2%) and IAA (41.0% vs 0%) were increased among ICA-positive siblings relative to ICA-positive parents (p<0.001). The siblings had higher ICA titers (p<0.001) than the parents but tended to have lower GADA levels (p=0.12). IA-2A levels did not differ between the two groups. IA-2A levels explained a higher proportion of the ICA reactivity in the siblings than in the parents (44% vs 12%, p=0.004), and GADA levels had the same tendency (27% vs 10%, p=0.11). In a multiple regression analysis, GADA and IA-2A were found to explain together 16% of the ICA reactivity in parents and 49% in siblings (p=0.003 for the difference). CONCLUSIONS: These results indicate that the increased frequency of additional diabetes-associated autoantibodies in ICA-positive siblings when compared to their ICA-positive parents may reflect the increased risk of progression to clinical type 1 diabetes previously reported in young ICA-positive relatives. We conclude that ICA immunofluorescence is not only due to GADA and IA-2A, but there are other additional antigens contributing to the ICA reactivity. Antibodies to such antigens appear to be more common among adults than in children. PMID- 11921418 TI - Changes in endoneurial blood flow, motor nerve conduction velocity and vascular relaxation of epineurial arterioles of the sciatic nerve in ZDF-obese diabetic rats. AB - BACKGROUND: We have previously reported that in streptozotocin-induced diabetic rats, reduction in endoneurial blood flow (EBF) and impairment of acetylcholine mediated vascular relaxation of arterioles that provide circulation to the sciatic nerve precedes slowing of motor nerve conduction velocity (MNCV). However, in animal models of type 2 diabetes it is unknown whether slowing of MNCV is accompanied by vascular dysfunction. METHODS: Using ZDF-lean and ZDF obese diabetic rats we examined whether diabetes-induced slowing in MNCV was associated with a reduction in EBF and impaired vascular relaxation in epineurial arterioles of the sciatic nerve. We measured MNCV in the sciatic nerve using a non-invasive procedure, and sciatic nerve nutritive blood flow using microelectrode polarography. In vitro videomicroscopy was employed to quantify arteriolar diameter responses to acetylcholine in arterioles overlying the sciatic nerve. RESULTS: MNCV and EBF in hyperglycemic (4-week duration) ZDF-obese diabetic rats were significantly decreased by 30% and 63%, respectively, compared to age-matched ZDF-lean rats. Acetylcholine elicited a dose-dependent dilation of epineurial vessels from ZDF-lean and ZDF-obese diabetic rats, although acetylcholine-induced dilation was significantly reduced in ZDF-obese diabetic rats. Determination of markers of oxidative stress provided ambiguous results. Superoxide levels were increased in epineurial vessels from ZDF-obese diabetic rats. Lens glutathione levels were decreased and serum thiobarbituric acid reactive substances increased in ZDF-obese diabetic rats but sciatic nerve conjugated diene and glutathione levels were not significantly different compared to ZDF-lean rats. CONCLUSIONS: Diabetes causes a reduction in EBF and impairment in vascular relaxation in epineurial vessels in ZDF-obese diabetic rats. This impaired vascular response is associated with neural dysfunction. PMID- 11921420 TI - Serum IFN-gamma and IL-10 levels are associated with disease progression in non obese diabetic mice. AB - BACKGROUND: The goal of the present study was to determine whether cytokines in the peripheral blood of naive NOD mice correlate with the disease process and thereby would provide a marker for monitoring disease activity. METHODS: Female NOD mice (5, 10 and 14-16 weeks of age) were investigated in a cross-sectional study. In the group of 14-16-week-old mice, non-diabetic and diabetic mice were analysed as different subgroups. The Th1 cytokine (IFN-gamma) and the Th2 cytokine (IL-10) were quantified in serum by sandwich enzyme-linked immunosorbent assay (ELISA). Pancreatic mRNA for IFN-gamma and IL-10 was determined by reverse transcriptase-polymerase chain reaction (RT-PCR) from the same animals. RESULTS: Serum levels of IFN-gamma were initially low but increased with age in NOD mice, reaching the highest levels at diabetes onset (p<0.002 compared to 10 weeks). A similar rise was noted in IFN-gamma gene expression in pancreatic lesions. In contrast, an early peak of serum IL-10 levels was observed in non-diabetic NOD mice (10 weeks) at a stage where non-destructive insulitis occurs. With increasing age a continuous loss of IL-10 until progression towards diabetes was observed. The pancreatic IL-10 mRNA expression correlated with serum IL-10 changes. As a consequence, the ratio of IFN-gamma/IL-10, reflecting the Th1/Th2 balance in the serum, was significantly increased in diabetic compared to non diabetic NOD mice (p<0.005). CONCLUSION: These results demonstrate, for the first time, that an increased Th2 pattern in the non-diabetic stage preceding a Th1 shift is associated with the development of diabetes in naive NOD mice. Serum cytokines correlate with disease progression and pancreatic cytokine expression during prediabetes. Soluble cytokines measured in the periphery are therefore promising surrogate markers of diabetes development. PMID- 11921419 TI - Prevalence and fate of type 1 diabetes-associated autoantibodies in cord blood samples from newborn infants of non-diabetic mothers. AB - BACKGROUND: Cord blood samples were collected from 1002 consecutive births at Turku University Hospital to study the prevalence and fate of type 1 diabetes associated autoantibodies in newborn infants of unaffected mothers. METHODS: The samples were analysed for cytoplasmic islet cell antibodies (ICA), autoantibodies to the 65 kD isoform of glutamic acid decarboxylase (GADA), autoantibodies to the protein tyrosine phosphatase related IA-2 antigen (IA-2A), insulin autoantibodies (IAA) and HLA DQB1 genotypes. RESULTS: ICA were detected in 27 cord blood samples (2.7%), with a median of 6 (range 4-34) JDF units. GADA were found to be positive (> or =6.6 RU) in six samples (0.6%), with a median of 66 (range 19-125) RU. IA 2A (> or =0.43 RU) were observed in three samples (0.3%), with a median of 1.3 (range 0.8-57) RU, while only one cord blood sample (0.1%) tested positive for IAA (> or =1.56 nU/ml) with a value of 5.4 RU. Maternal or gestational age, sex and birth weight of the infant were not related to antibody prevalence or titres. Altogether there were 29 infants with antibody positivity in their cord blood (2.9%). Five of these (0.5%) tested positive for two antibodies (ICA and GADA), and one was positive for all four antibodies measured. All nine infants from whom follow-up samples were available became antibody negative by the age of 15 months, and in all but one case inverse seroconversion occurred by the age of 9 months. CONCLUSIONS: Around 3% of infants of non-diabetic mothers in Finland have diabetes-associated autoantibodies at birth, and these antibodies disappear at the latest by the age of 15 months. PMID- 11921422 TI - Current literature in diabetes. PMID- 11921421 TI - Effects of angiotensin II receptor antagonist on endothelial vasomotor function and urinary albumin excretion in type 2 diabetic patients with microalbuminuria. AB - BACKGROUND: Microalbuminuria is associated with dysfunction of the vascular endothelium in patients with diabetes mellitus. The objective of the present study was to determine whether treatment with losartan at a dose sufficient to lower urinary albumin excretion was accompanied by an improvement in endothelial function in type 2 diabetic patients with microalbuminuria. METHODS: Endothelial function was measured in 80 type 2 diabetic patients with microalbuminuria and 68 non-diabetic controls using high-resolution vascular ultrasound. The diabetic patients were randomised to receive either losartan 50 mg daily or placebo in a 6 month double-blind study. Urinary albumin excretion and endothelial function were assessed at baseline, 3 and 6 months. RESULTS: Both endothelium-dependent (p<0.01) and -independent vasodilation (p<0.01) were significantly impaired in diabetic patients with or without history of hypertension compared to the non diabetic controls. At baseline, the losartan- and placebo-treated groups were comparable in their clinical characteristics. Blood pressure did not change significantly in either group throughout the study. Urinary mean albumin excretion rate (MAER) decreased in the losartan-treated group (p<0.01) whereas an increase was observed in the placebo group (p<0.05). At 6 months, the losartan treated group had significantly lower MAER than the placebo-treated group [54.5 (58.3) vs 78.5 (100.5) microg/min, p<0.05; median (interquartile range)]. No significant differences were found in endothelium-dependent or -independent vasodilation. CONCLUSIONS: Type 2 diabetic patients with microalbuminuria have impaired endothelium-dependent and -independent vasodilation. Treatment with low dose losartan is sufficient to reduce microalbuminuria in these patients without alteration in endothelial function and systemic blood pressure. PMID- 11921423 TI - An introduction to new advances in diabetes. AB - Diabetes is a potentially devastating disease with a high morbidity and mortality. There is an excess risk of both microvascular and macrovascular complications with diabetes [1]. Recent studies have emphasised and illustrated how we might be able to limit these diabetic complications. The Diabetes Control and Complications Trial (DCCT) and the United Kingdom Prospective Diabetes Study (UKPDS) found that improved control of blood glucose reduced the risk of major diabetic eye disease by 25%, serious deterioration of vision by nearly 50%, and early kidney damage by 33%. Other studies including the UKPDS have demonstrated the importance of blood pressure control and reduced cholesterol in addition to the use of aspirin in limiting progression of macrovascular disease. Diabetes is no longer viewed as a disease of sugar alone; a more holistic approach is required if our patients are to benefit from the information we have acquired through these recent studies. Some of the most recent developments in the field are presented in this review. PMID- 11921424 TI - Therapeutic targets in the management of Type 1 diabetes. AB - For historical reasons, diabetes has long been linked with blood and urine glucose control, partly because these were clearly linked to acute symptoms, and partly because glucose became measurable around 200 years ago. Today it is recognized that there is far more to diabetes than simply monitoring symptoms and blood glucose. Intensive management has an impact on the quality of life. Late complications have their own risk factors and markers. Monitoring and early detection of these risk factors and markers can lead to changes in treatment before tissue damage is too severe. Accordingly, professionals now find themselves monitoring a range of adverse outcomes, markers for adverse outcomes, risk factors and risk markers for microvascular and arterial disease, acute complications of therapy, and the care structures needed to deliver this. Adverse outcomes lend themselves to targets for complication control in populations, and markers of adverse outcomes (such as retinopathy and raised albumin excretion rate) in treatment cohorts. Surveillance systems will have targets for yearly recall and review of early complications. Metabolic (surrogate) outcomes can be monitored in individual patients, but monitoring is only of value in so far as it guides interventions, and this requires comparison to some intervention level or absolute target. Even for blood glucose control this is not easy, for conventional measures such as glycated haemoglobin have their own problems, and more modern approaches such as post-prandial glucose levels are controversial and less convenient to measure. In many people with type 1 diabetes targets for blood pressure, LDL cholesterol, and serum triglycerides will also be appropriate, and need to be part of any protocol of management. PMID- 11921425 TI - Diabetes management in the new millennium using insulin pump therapy. AB - Current goals of therapy of type 1 and 2 diabetes are to achieve near normal glycemia, minimize the risk of severe hypoglycemia, limit excessive weight gain, improve quality of life and delay or prevent late vascular complications. As discussed in this review, insulin pump or continuous subcutaneous insulin infusion (CSII) therapy provides a treatment option that can dramatically aid in achieving all of these goals. In comparison to multiple daily injections (MDI), CSII uses only rapid-acting insulin, provides greater flexibility in timing of meals and snacks, has programmable basal rates to optimize overnight glycemic control, can reduce the risk of exercise-induced hypoglycemia, and enhances patients' ability to control their own diabetes. Most important, in adults and adolescents with type 1 diabetes, CSII has been shown to lower HbA(1c) levels, reduce the frequency of severe hypoglycemia and limit excessive weight gain versus MDI without increasing the risk of diabetic ketoacidosis. Similarly positive results are being seen with CSII in adults with type 2 diabetes. The effectiveness of CSII and improvements in pump technology have fueled a dramatic increase in the use of this therapy. Practical guidelines are presented for selection of patients, initiation of treatment, patient education, follow-up assessments and troubleshooting. The recent introduction of methods for continuous glucose monitoring provides a new means to optimize the basal and bolus capabilities of CSII and offers the hope of the development of a feedback controlled artificial pancreas. PMID- 11921426 TI - Insulin analogues: have they changed insulin treatment and improved glycaemic control? AB - To improve insulin therapy, new insulin analogues have been developed. Two fast acting analogues with a more rapid onset of effect and a shorter duration of action combined with a low day-to-day variation in absorption rate are now available. Despite this favourable time-action profile most studies have not been able to show any improvement in overall glycaemic control with the fast-acting analogues. A reduced post-prandial increase in blood glucose has been found in all studies, whereas between 3 and 5 h after the meal and during the night an increased blood glucose level is the normal course. This is probably the main explanation for the absence of improvement in overall glycaemic control when compared with regular human insulin. A tendency to a reduction in hypoglycaemic events during treatment with fast-acting analogues has been observed in most studies. Recent studies have indicated that NPH insulin administered several times daily at mealtimes can improve glycaemic control without increasing the risk of hypoglycaemia. The fast-acting analogues are now also available as insulin mixed with NPH. Insulin glargine is a new long-acting insulin which is soluble and precipitates after injection, resulting in a long half-life with a residual activity of about 50% 24 h after injection. Insulin glargine is a peakless insulin and studies in both type 1 and type 2 diabetic patients indicate that glargine improves fasting blood glucose control and reduces the incidence of nocturnal hypoglycaemia. Surprisingly, the new fast-acting analogues have not achieved the expected commercial success, which emphasises the need for new strategies for basal insulin supplementation, exercise, diet and blood glucose monitoring. PMID- 11921427 TI - Development of oral insulin: progress and current status. AB - NOBEX oral drug delivery technology is centered on modification of peptides, proteins and small organic molecules by attachment of one or more amphiphilic oligomers. Attachment of these oligomers results in stability to enzymatic degradation, improved solubility to allow optimized formulation, and modification of pharmacology to prolong circulating half-life and activity. NOBEX has applied this technology to insulin, creating an orally absorbed, bioactive conjugate which is safe and rapidly absorbed and which demonstrates dose-dependent, glucose lowering effects in animal models, healthy volunteers and type 1 diabetic patients. Results of recently completed and ongoing studies in both type 1 and type 2 diabetes suggest a promising role for conjugated insulin in the management of fasting and post-prandial hyperglycemia. Orally administered conjugated insulin is delivered first to the liver through the portal circulation, similar to the physiological route of insulin secretion in non-diabetic individuals. Potential benefits from this route of insulin delivery include improved disease management and reduction of long-term complications of diabetes. Further studies with oral conjugated insulin are planned in both type 1 and type 2 diabetic patients. PMID- 11921428 TI - The evolving role of oral insulin in the treatment of diabetes using a novel RapidMist System. AB - The inability of subcutaneous (sc) insulin to effectively, safely and painlessly control postprandial glucose levels has encouraged the exploration of alternate methods of insulin delivery. Recently, a novel drug delivery system, based on a unique liquid aerosol formulation, has been developed. This system allows precise insulin dose delivery via a simple, cosmetically acceptable metered dose inhaler in the form of fine aerosolized droplets directed into the mouth. The system introduces a fine-particle aerosol at high velocity into the patient's breath; the mouth deposition is dramatically increased compared with conventional technology. This oral aerosol formulation is rapidly absorbed through the buccal mucosal lining and in the oropharynx regions, and it provides the plasma insulin levels necessary to control postprandial glucose rise in diabetic patients. This novel, pain-free, oral insulin formulation has a critical series of attributes: rapid absorption, a simple (user-friendly) administration technique, precise dosing control (comparable to injection within one unit), and bolus delivery of drug. This review describes the recent results of clinical studies (in type 1 and type 2 diabetic patients) by comparing the efficacy of Oralin (oral insulin spray) versus sc injected insulin and placebo arms. A simplified means for prandial insulin delivery, such as that offered by this technique, will significantly reduce the incidence of key complications by allowing increased patient compliance for consistent drug administration in order to regulate patients' blood glucose levels. PMID- 11921429 TI - Making 'sense' of diabetes: using a continuous glucose sensor in clinical practice. AB - Home blood glucose monitoring provides only a snapshot picture of prevailing glucose values. Continuous glucose monitoring allows identification of patterns of glucose levels for up to 72 h with details of excursions above and below target levels and thus may be a valuable adjunct tool in diabetes management. We have used the MiniMed Continuous Glucose Monitoring System (CGMS) in ten individuals and have uncovered a significant rate of hypoglycaemia that was previously undetected by conventional means. Preliminary evidence suggests that the novel technology may allow clinicians to 'personalise' intensive insulin therapy with improvement in glycaemic control. PMID- 11921430 TI - Glucose monitoring by reverse iontophoresis. AB - Glucose can be extracted through intact skin by electro-osmotic flow (a process called 'reverse iontophoresis') upon the application of a low-level electrical current. Recently we have combined iontophoretic extraction with an in situ glucose sensor in a device called the GlucoWatch biographer. Clinical results with this device show close tracking of blood glucose over a range of 2.2 to 22.2 mmol/l for up to 12 h using a single blood glucose value as calibration. The biographer readings lag behind blood glucose values by an average of 18 min. An analysis of data from 92 diabetic subjects in a controlled clinical setting shows a linear relationship (r=0.88) between GlucoWatch biographer readings and blood glucose. The mean absolute relative difference between the two measurements was 15.6% and more than 96% of the data fell in the (A+B) regions of the Clarke error grid. Similar results have been obtained from subjects using the GlucoWatch biographer in an uncontrolled home environment. The automatic, frequent, and non invasive measurements obtained with the GlucoWatch biographer provide substantially more information about glucose levels than do the current fingerstick methods. This information can be used for improved decisions about all aspects of diabetes management. PMID- 11921431 TI - Are all glitazones the same? AB - This supplement focuses on the benefits of targeting insulin resistance through therapy with a new class of oral antidiabetic agents, the thiazolidinediones (TZDs) or 'glitazones'. There are important differences between the three TZD class members that warrant discussion to enable physicians to make rational and informed therapeutic choices between the agents. Overall the TZDs appear to be similar in their effects on blood glucose, as all class members have demonstrated effective glycaemic control, both as monotherapy and in combination with sulphonylureas, metformin or exogenous insulin. The safety profiles of the three agents are more diverse, with what appear to be 'TZD class effects', (probably mediated via activation of peroxisome proliferator-activated receptor gamma [PPAR gamma]) and 'TZD-specific effects', which are unique to each agent and may be a consequence of differing chemical structures. While rosiglitazone and pioglitazone share some class effects with troglitazone, they have several characteristics that define them as unique agents. By tackling the control of type 2 diabetes through direct effects on insulin resistance, the TZDs represent an important new therapeutic tool for healthcare professionals. PMID- 11921433 TI - The mode of action of thiazolidinediones. AB - The thiazolidinediones (TZDs) or 'glitazones' are a new class of oral antidiabetic drugs that improve metabolic control in patients with type 2 diabetes through the improvement of insulin sensitivity. TZDs exert their antidiabetic effects through a mechanism that involves activation of the gamma isoform of the peroxisome proliferator-activated receptor (PPAR gamma), a nuclear receptor. TZD-induced activation of PPAR gamma alters the transcription of several genes involved in glucose and lipid metabolism and energy balance, including those that code for lipoprotein lipase, fatty acid transporter protein, adipocyte fatty acid binding protein, fatty acyl-CoA synthase, malic enzyme, glucokinase and the GLUT4 glucose transporter. TZDs reduce insulin resistance in adipose tissue, muscle and the liver. However, PPAR gamma is predominantly expressed in adipose tissue. It is possible that the effect of TZDs on insulin resistance in muscle and liver is promoted via endocrine signalling from adipocytes. Potential signalling factors include free fatty acids (FFA) (well known mediators of insulin resistance linked to obesity) or adipocyte-derived tumour necrosis factor-alpha (TNF-alpha), which is overexpressed in obesity and insulin resistance. Although there are still many unknowns about the mechanism of action of TZDs in type 2 diabetes, it is clear that these agents have the potential to benefit the full 'insulin resistance syndrome' associated with the disease. Therefore, TZDs may also have potential benefits on the secondary complications of type 2 diabetes, such as cardiovascular disease. PMID- 11921432 TI - Insulin resistance in type 2 diabetes: role of fatty acids. AB - Insulin resistance is one of the key factors responsible for hyperglycaemia in type 2 diabetes and can result in a number of metabolic abnormalities associated with cardiovascular disease (insulin resistance syndrome), even in the absence of overt diabetes. The mechanisms involved in the development of insulin resistance are multifactorial and are only partly understood, but increased availability of free fatty acids (FFAs) is of particular importance for the liver and skeletal muscle. The role of FFAs in type 2 diabetes is most evident in obese patients who have several abnormalities in FFA metabolism. Because of a mass effect, the release of FFAs from the total adipose tissue depot to the blood stream is increased and the high concentration of circulating FFAs impairs muscle uptake of glucose by competitive inhibition. In upper-body obesity, which predisposes individuals to type 2 diabetes, the rate of lipolysis is accelerated in visceral adipose tissue. This results in a selective increase in FFA mobilisation to the portal vein, which connects visceral fat to the liver. A high 'portal' FFA concentration has undesirable effects on the liver, resulting in dyslipidaemia, hyperinsulinaemia, hyperglycaemia and hepatic insulin resistance. Recently, a new class of antidiabetic agents, the thiazolidinediones (TZDs) or 'glitazones' has been developed. A prominent effect of these agents is the lowering of circulating FFA levels and it is believed, but not yet proven, that this interaction with FFAs constitutes a major mechanism behind the glucose-lowering effect of the TZDs. PMID- 11921434 TI - Differentiating members of the thiazolidinedione class: a focus on efficacy. AB - The thiazolidinediones (TZDs) or 'glitazones' are a new class of drug used for the treatment of type 2 diabetes. Although their precise mechanism of action is not known, TZDs target insulin resistance directly and thus tackle an underlying cause of the disease. Two TZDs are indicated for use in type 2 diabetes in the USA, pioglitazone and rosiglitazone. A third, troglitazone, has been associated with significant hepatotoxicity and has been withdrawn from use. In clinical trials, all three TZDs effectively lower blood glucose levels as monotherapy and in combination therapy with sulfonylureas, metformin and insulin. To date, head to-head comparative studies with these agents have not been performed. It is difficult, therefore, to make direct comparisons of their efficacy since other variables, including baseline glucose levels and study design, can have a significant impact on treatment outcome. Despite this and in light of unique safety issues characterized with certain TZDs, it is useful to look closely at the efficacy data for these agents. It is not sufficient to assume that 'all glitazones are the same' because the studies have not yet been done to support this statement. This article will review what is known about the relative efficacy of the TZDs. PMID- 11921436 TI - Where thiazolidinediones will fit. AB - Individuals with type 2 diabetes have two defects: insulin resistance, which occurs in the first stages of disease progression, and pancreatic beta-cell failure, which occurs later in the disease. Insulin resistance is the major pathological defect. During the course of the disease, insulin levels are initially elevated to compensate for the increased insulin resistance and then decline as the disease progresses and beta-cells become less responsive. It is necessary to change antidiabetic therapies to address this progression. Current management of type 2 diabetes follows a stepwise treatment program of diet and exercise, monotherapy with oral antidiabetic agents, combination oral therapy and, ultimately, combination therapy with insulin to control blood glucose levels. While control of blood glucose will reduce the risk of microvascular complications, such as microalbuminuria and retinopathy, the incidence of macrovascular complications is not significantly reduced. The introduction of the thiazolidinediones (TZDs) or 'glitazones', a class of agents that offer effective glycemic control and work through the reduction of insulin resistance, offers a new strategy in the management of this condition. These agents have beneficial effects on the pancreatic beta-cell and, in addition, may have potential benefits on the macrovascular complications that commonly occur in these patients. PMID- 11921435 TI - Differentiating members of the thiazolidinedione class: a focus on safety. AB - Troglitazone, rosiglitazone and pioglitazone are members of the thiazolidinedione (TZD) class - antidiabetic agents that have proven efficacy in the treatment of patients with type 2 diabetes. All three agents are believed to mediate their effects via activation of the gamma isoform of the peroxisome proliferator activated receptor (PPAR gamma). Despite this common mechanism of action, they all have unique chemical structures and receptor-binding affinities, and consequently, in addition to the class effects (probably mediated through PPAR gamma), each TZD has a unique safety profile. Side effects have been categorized as unique to individual TZDs, or common to the class of drug. Of the unique effects, the best characterized is hepatotoxicity, which has been associated specifically with troglitazone to date. Studies with rosiglitazone and pioglitazone indicate that hepatotoxicity is not a class effect. Further differences in the safety profiles of these agents arise because the oxidative metabolism for each agent occurs by distinct cytochrome pathways: troglitazone and pioglitazone involve CYP 3A4 and CYP 2C8 whereas rosiglitazone is principally metabolized by CYP 2C8. CYP 3A4 is involved in the metabolism of over 150 drugs, hence the potential for drug interactions with troglitazone and pioglitazone is much greater than with rosiglitazone. Class effects include edema, slight reductions in hemoglobin and hematocrit (due to hemodilution), weight gain and alterations in plasma lipid profiles. This article considers safety data obtained from both clinical trials and clinical practice as a means of differentiating among troglitazone, rosiglitazone and pioglitazone. PMID- 11921438 TI - Differential gel exposure, a new methodology for the two-dimensional comparison of protein samples. AB - We describe a novel methodology for the comparison of protein samples called differential gel exposure (DifExpo). This method is based on the coelectrophoresis on a two-dimensional (2-D) gel of two protein samples. The samples are differentiated from each other by in vivo radiolabelling, using (14)C and (3)H-isotopes. After 2-D separation and transfer on a polyvinylidene difluoride membrane, the (3)H/(14)C ratio of each protein spot is determined by exposure to two types of imaging plates, one sensitive to (14)C and the other to both (14)C and (3)H. We showed that DifExpo allows us to compare the cellular levels of several hundred proteins of the yeast proteome. Its sensitivity is comparable to silver staining. We also showed that it can be used to investigate changes in the rate of synthesis of individual proteins. PMID- 11921439 TI - Functional proteomics using chromophore-assisted laser inactivation. AB - Proteins are the molecules that fulfil most cellular functions and represent over 90% of drug targets in the market. Chromophore-assisted laser inactivation (CALI) provides a timely and locally restricted protein inactivation and has proven to specifically destroy protein function using dye-coupled ligands and laser irradiation. CALI involves the generation of short-lived radicals thus limiting the radius of covalent modifications to spatially restricted sites on the target molecule. A transient functional inactivation occurs if the radicals modify amino acids of the target protein that are responsible for function. Here we show specific inactivation of several protein targets, that are members of relevant signal transduction pathways. For each of these targets, simple and high throughput screening-scaleable assays have been developed, making it possible to quantify the observed inactivation. Activities of target proteins have been addressed in cell-free as well as cell-based assays employing human primary and tumor-derived cell lines. In all cases, at least 50% inactivation was achieved. The data presented here demonstrate that CALI is a highly versatile tool for validating disease relevant targets at the protein level. This approach also takes into account post-translational modifications like phosphorylation, glycosylation or acylation, thereby enlarging its applicability for many different types of targets. PMID- 11921440 TI - Fluorophore-assisted light inactivation: a high-throughput tool for direct target validation of proteins. AB - To exploit advances in proteomics for drug discovery, high-throughout methods for target validation that directly address the cellular roles of proteins are required. To do this, we have characterized fluorophore-assisted light inactivation (FALI) which uses coherent or diffuse light targeted by fluorescein labeled probes to inactivate specific proteins. We have shown that it is spatially restricted and tested its efficacy in living cells. FALI is efficient using conventional antibodies and single chain variable fragment phage display antibodies (that are compatible with high-throughput applications). We have shown that singlet oxygen is one of the major components required for FALI-mediated damage. The half-maximal radius of damage is approximately 40 A. FALI causes the specific loss of function of beta 1 integrin in HT-1080 fibrosarcoma cells resulting in a reduction in invasiveness. The efficacy of diffuse light sources (such as a desk lamp) with FALI to inactivate many samples in parallel provides an inexpensive, high-throughput method of wide general applicability for functional proteomics. PMID- 11921441 TI - Functional proteomics using microchannel plate detectors. AB - We describe the development of a novel detection system used for the functional imaging of proteins separated on electrophoretic gels. A microchannel plate detector is used here for real-time imaging of low levels of tritiated protein separated by two-dimensional (2-D) electrophoresis. The system employs radioisotope-free, low noise microchannel plates originally developed for photon counting in X-ray astronomy. Using the detector configuration described here, proteins were resolved on mini gels by either one or two-dimensional electrophoresis, transferred onto polyvinylidene difluoride membranes and directly imaged. Tritiated diisopropylfluorophosphate (DFP) was used as a selective label for the serine hydrolase class of enzymes and their distribution in the central nervous system was examined. This survey revealed approximately 24 protein spots by 2-D electrophoresis. We also investigated the relative sensitivity of these proteins towards DFP and found the peptidase, acylpeptide hydrolase to be the most sensitive brain protein towards this reagent. Using a number of different tritiated standards, it was found that the system can image as little as 0.1 Bq/mm(2) of tritium corresponding to 320 attomol of DFP labelled protein/mm(2). Moreover, the system has a wide dynamic range (>10(6)) allowing samples of high and low activity to be quantified on the same gel. PMID- 11921442 TI - A model of random mass-matching and its use for automated significance testing in mass spectrometric proteome analysis. AB - A rapid and accurate method for testing the significance of protein identities determined by mass spectrometric analysis of protein digests and genome database searching is presented. The method is based on direct computation using a statistical model of the random matching of measured and theoretical proteolytic peptide masses. Protein identification algorithms typically rank the proteins of a genome database according to a score based on the number of matches between the masses obtained by mass spectrometry analysis and the theoretical proteolytic peptide masses of a database protein. The random matching of experimental and theoretical masses can cause false results. A result is significant only if the score characterizing the result deviates significantly from the score expected from a false result. A distribution of the score (number of matches) for random (false) results is computed directly from our model of the random matching, which allows significance testing under any experimental and database search constraints. In order to mimic protein identification data quality in large-scale proteome projects, low-to-high quality proteolytic peptide mass data were generated in silico and subsequently submitted to a database search program designed to include significance testing based on direct computation. This simulation procedure demonstrates the usefulness of direct significance testing for automatically screening for samples that must be subjected to peptide sequence analysis by e.g. tandem mass spectrometry in order to determine the protein identity. PMID- 11921443 TI - A strategy for mapping and neutralizing conformational immunogenic sites on protein therapeutics. AB - Antibodies are highly specific recognition molecules which are increasingly being applied to target therapy in patients. One type of developmental antibody-based therapy is antibody directed enzyme prodrug therapy (ADEPT) for the treatment of cancer. In ADEPT, an antibody specific to a tumor marker protein delivers a drug activating enzyme to the cancer. Subsequent intravenous administration of an inactive prodrug results in drug activation and cytotoxicity only within the locale of the tumor. Pilot clinical trials with chemical conjugates of the prodrug activating enzyme carboxypeptidase G2 (CPG2) chemically conjugated with an antibody to and carcinoembryonic antigen (CEA), have shown that CPG2-mediated ADEPT is effective but limited by formation of human antibodies to CPG2 (HACA). We have developed a recombinant fusion protein (termed MFE-CP) of CPG2 with an anti-CEA single chain Fv antibody fragment and we have developed methods to address the immunogenicity of this therapeutic. A HACA-reactive discontinuous epitope on MFE-CP was identified using the crystal structure of CPG2, filamentous phage technology and surface enhanced laser desorption/ionization affinity mass spectrometry. This information was used to create a functional mutant of MFE-CP with a significant reduction (range 19.2 to 62.5%, median 38.5%) in reactivity with the sera of 11 patients with post-therapy HACA. The techniques described here are valuable tools for identifying and adapting undesirable immunogenic sites on protein therapeutics. PMID- 11921444 TI - Primary skin fibroblasts as human model system for proteome analysis. AB - Elucidation of cellular processes and their changes at the level of protein expression and post-translational modification patterns may allow identification of novel proteins and thereby mechanisms involved in the pathogenesis of multigenic diseases. The aim of this study was to test cultured, nontransformed primary fibroblasts derived from human skin biopsies as a suitable model system for proteome analysis. Therefore soluble protein fractions were separated on several overlapping ultrazoom gels covering the pH range from 3.5-9. Correlation analysis of gel-pairs revealed a highly reproducible protein expression pattern within (intra-assay) and between (inter-assay) independent experiments of a single fibroblast cell line (intra-cell line comparison). Spot intensity variations were less than a factor of two for more than 80% of identical spots. In addition, inter-cell line comparison exhibits no significant variations in spot intensities. To achieve further improvements in reproducibility we generated master gels for each pH range by combining averaged spot information derived from two different cell lines each analysed by two independent experiments using the raw master gel algorithm of the Z3 image analysis software. The resulting reference images of primary human fibroblasts provided a basis for investigating regulation by extracellular stimuli and drugs as well as their alterations in patients with different diseases. PMID- 11921445 TI - Towards complete analysis of the platelet proteome. AB - Platelets exert a crucial function in haemostasis, wound repair, and the formation of vascular plugs, underlying thrombotic diseases such as stroke and myocardial infarction. Analysis of platelet biochemistry is largely dependent on protein analysis as platelets are anucleated cells providing little analytical target for DNA or RNA based strategies. Here we present data from our analysis of the human platelet proteome, the entire set of proteins building a platelet at a given point in time. Proteins were separated by two-dimensional electrophoresis (2-DE) using broad and narrow range pH gradients in the isoelectric focusing step. Consequently, a high-resolution 2-DE proteome map has been generated that comprises approximately 2300 different protein features. From the 536 protein features detected in the 4-5 pI range 284 features were identified by electrospray ionisation time of flight tandem mass spectrometry. These 284 proteins originate from 123 different open reading frames. This includes the five human proteins KIAA0193, KIAA0573, KIAA0830, WUGSC:H_DJ0777O23 protein, and cytokine receptor related protein 4, all isolated for the first time. The data are discussed with regard to proteome characteristics, protein function, and the high prevalence of signalling molecules. This study contributes to a more thorough and holistic understanding of platelet biology, helping to build the basis for future identification of new drug targets and therapeutic strategies. PMID- 11921446 TI - Proteomic analysis of differential protein expression in human nasopharyngeal carcinoma cells induced by NAG7 transfection. AB - Nasopharyngeal carcinoma (NPC) is a commonly occurring tumor in southern China and south east Asia. A genetic factor has now been recognized to be associated with this cancer. A new gene, named NAG7, was cloned from the common minimal deletion region in 3p25.3-26.3. In order to investigate the function of NAG7 gene, proteomic methods were used to find and identify the differential proteins and expected to elucidate the mechanism of NAG7. The NAG7 eukaryotic expression vector was constructed and transfected into NPC cell line HNE1 with liposome. Twenty-two differential protein spots in transfected cells were found significant and reproducible using high-resolution two-dimensional electrophoresis. Nine proteins that were up-regulated and seven proteins that were down-regulated were identified by matrix assisted laser desorption/ionization time of flight mass spectrometry and database analysis. These proteins included growth arrest specific protein, DNA binding protein, caspase 6, pinch protein and ras-related protein rab-36, which are involved in cell cycling, transcription regulation, signaling pathways and apoptosis. NAG7 may exert its functions by mediating differential expression of these proteins. PMID- 11921447 TI - Immunoproteomics of Helicobacter pylori infection and relation to gastric disease. AB - The Gram negative bacterium Helicobacter pylori is a human pathogen which infects the gastric mucosa and causes an inflammatory process leading to gastritis, ulceration and cancer. A systematic, proteome based approach was chosen to detect candidate antigens of H. pylori for diagnosis, therapy and vaccine development and to investigate potential associations between specific immune responses and manifestations of disease. Sera from patients with active H. pylori infection (n = 24), a control group with unrelated gastric disorders (n = 12) and from patients with gastric cancer (n = 6) were collected and analyzed for the reactivity against proteins of the strain HP 26695 separated by two-dimensional electrophoresis. Overall, 310 antigenic protein species were recognized by H. pylori positive sera representing about 17% of all spots separated. Out of the 32 antigens most frequently recognized by H. pylori positive sera, nine were newly identified and 23 were confirmed from other studies. Three newly identified antigens which belong to the 150 most abundant protein species of H. pylori, were specifically recognized by H. pylori positive sera: the predicted coding region HP0231, serine protease HtrA (HP1019) and Cag3 (HP0522). Other antigens were recognized differently by sera from gastritis and ulcer patients, which may identify them as candidate indicators for clinical manifestations. The data from these immunoproteomic analyses are added to our public database (http://www.mpiib berlin.mpg.de/2D-PAGE). This platform enables one to compile many protein profiles and to integrate data from other studies, an approach which will greatly assist the search for more immunogenic proteins for diagnostic assays and vaccine design. PMID- 11921448 TI - Characterisation by proteomics of peribacteroid space and peribacteroid membrane preparations from pea (Pisum sativum) symbiosomes. AB - The legume Rhizobium symbiosis leads to the formation of a new compartment in the plant cell, the symbiosome. This compartment harbours the bacteroids surrounded by a peribacteroid membrane (PBM) originating from the plant plasma membrane. The PBM and the space between the PBM and the bacteroid membrane, called peribacteroid space (PS), mediate the exchange of metabolites between the symbionts. Proteome analysis was used as an approach to characterise the proteins in the PBM and the PS. A standard differential centrifugation procedure including a Percoll gradient was used for symbiosome isolation from pea root nodules. Proteins in the PBM and PS fractions obtained from the symbiosomes were separated by two-dimensional gel electrophoresis, and 89 spots were analysed by tandem mass spectrometry. The proteins of 46 spots could be identified by database search. The results showed that PS and even PBM preparations from pea symbiosomes always contain abundant amounts of bacteroid proteins as a contaminate. Interestingly, in addition to a few PS/PBM proteins a number of endomembrane proteins (less likely representing a contaminate), including V-ATPase, BIP, and an integral membrane protein known from COPI-coated vesicles, were found in the PBM fraction, supporting the role of the endomembrane system in PBM biogenesis. PMID- 11921452 TI - Effects of socioeconomic status on presentation with acute lower respiratory tract disease in children in Salvador, Northeast Brazil. AB - Two different socioeconomic groups of children with pneumonia were studied, and their clinical and demographic aspects were evaluated. The diagnosis of pneumonia was based on findings of cough and tachypnea, or on crackles on auscultation or on radiologically confirmed infiltrate. This was a prospective cross-sectional study conducted at the Professor Hosannah de Oliveira Pediatric Center, which cares for children of lower socioeconomic status (PHOPC), and at one private hospital which cares for children from middle to high socioeconomic status (Alianca Hospital, AH). Demographics and clinical differences were assessed by the Pearson chi-square test or Fisher's exact test as appropriate; means of continuous variables were compared by Mann-Whitney U-test. In a 26-month period, 3,431 cases were recruited. The 2,476 cases identified at the PHOPC were younger than the 955 identified at AH (2.2 +/- 2.3 vs. 4.5 +/- 3.1 years, P < 0.0001) and had higher scores for severity (3.5 +/- 1.5 vs. 2.7 +/- 1.7, P < 0.0001), duration of hospitalization (days) (10.9 +/- 12.1 vs. 6.2 +/- 7, P < 0.0001), frequency of tobacco smoker in the household (48% vs. 31%, P < 0.0001), cardiopathy (15.3% vs. 5.9%, P = 0.003), fever (44.4% vs. 36.3%, P = 0.0001), tachypnea (67.6% vs. 32.3%, P < 0.0001), crackles (69.5% vs. 64.9%, P = 0.02), somnolence (19.9% vs. 10.4%, P < 0.0001), malnutrition (13.7% vs. 5%, P < 0.0001), hospitalization rate (27.4% vs. 22.5%, P = 0.003), and death (0.9% vs. 0.1%, P = 0.009). However, other features were more frequent among AH cases: parent's university level of education (38.2% vs. 1.0%, P < 0.0001), underlying chronic illness (40.6% vs. 28.5%, P < 0.0001), asthma (62.7% vs. 50.8%, P = 0.01), rhinitis (9.2% vs. 0.4%, P < 0.0001), previous use of antibiotics (34.3% vs. 27.1%, P = 0.001), and wheezing (53.1% vs. 42.2%, P < 0.0001). Children of lower socioeconomic status have more serious lower respiratory tract disease, whereas children with pneumonia of middle to high socioeconomic status have more allergic diseases (rhinitis, asthma) and wheezing. PMID- 11921451 TI - Maternal history, sensitization to allergens, and current wheezing, rhinitis, and eczema among children in Costa Rica. AB - Little is known about the factors associated with asthma, allergic rhinitis, and eczema in Latin American countries. We investigated the relation between potential risk factors and current wheezing, allergic rhinitis, and eczema among 208 Costa Rican children aged 10-13 years participating in phase II of the International Study of Asthma and Allergies in Childhood (ISAAC). The geometric mean ( +/- SD) serum total IgE level of children with current wheezing was significantly higher than that of children without current wheezing (533.8 +/- 5.2 vs. 144.7 +/- 6.0 IU/mL, P < 0.01). In a multivariate analysis, a maternal history of asthma, skin test reactivity (STR) to house dust mites, and STR to Alternaria were significantly associated with current wheezing. Children who had a maternal history of asthma had 2.4 times higher odds of current wheezing than those without maternal history of asthma (95% CI for OR = 1.1-5.3). Sensitization to either house dust mite or Alternaria was associated with 3.3 times increased odds of current wheezing (95% CI for OR for STR to dust mite = 1.6-6.7; 95% CI for OR for STR to Alternaria = 1.1-11.0). In a multivariate analysis, STR to house dust mite and STR to cat dander were significantly associated with allergic rhinitis, and a maternal history of eczema and STR to dog dander were associated with eczema in the child. The interaction between familial factors and lifestyle changes resulting from social reforms implemented 60 years ago may explain the high prevalence of atopic diseases in Costa Rica. PMID- 11921453 TI - Circulating adhesion molecules in sera of asthmatic children. AB - Infiltration of cells into the lung in asthma is regulated by several expressions of cell adhesion molecules (CAMs) on cells present in the airways, and may play a role in the pathogenesis of bronchial asthma. We sought to evaluate the role of serum concentrations of the soluble forms of intercellular adhesion molecule-1 (sICAM-1), vascular cell adhesion molecule-1 (sVCAM-1), and E-selectin (sE selectin) in the control of disease activity in acute asthma. Circulating levels of sICAM-1, sVCAM-1, and sE-selectin in sera from 15 normal control subjects and from 20 allergic asthmatic children with acute exacerbations who had returned to stable condition were determined by using commercially available enzyme-linked immunosorbent assay kits. The mean concentration of serum sICAM-1 levels was significantly higher during an acute exacerbation of asthmatic children than in those with stable asthma (19.41 +/- 10.65 ng/mL vs. 13.46 +/- 5.44 ng/mL; P < 0.001) or in control subjects (9.83 +/- 2.02 ng/mL; P < 0.001). For sVCAM-1 and sE-selectin, the mean serum concentration of sVCAM-1 was slightly higher in children during an acute exacerbation asthma than when stable. However, the differences did not reach statistical significance. The mean serum concentrations of sVCAM-1 and sE-selectin in acute asthma or stable asthma were significantly higher than in control subjects. This study provides further evidence that serum concentrations of sICAM-1, sVCAM-1, and sE-selectin are increased in acute asthma. These findings further confirm that leukocyte endothelial adhesion plays a role in inflammatory airway disease. PMID- 11921454 TI - Platelet endothelial cell adhesion molecule-1 and capillary loading in premature infants with and without chronic lung disease. AB - It was our objective to quantify platelet endothelial cell adhesion molecule-1 (PECAM-1) by immunohistochemistry in control infants of 22-50 weeks postconceptual age, and to correlate it with varying degrees of neonatal chronic lung disease (CLD). We tested the hypothesis that the density of PECAM-1 staining will positively correlate with increasing gestational age (GA) and the inflammatory process in CLD. A library of postmortem lung tissue of infants receiving ventilator care was accessed. The population consisted of 35 control infants exposed briefly to oxygen and positive pressure ventilation, and 31 infants who were 23-30 weeks GA with mild to severe CLD. A monoclonal anti-human PECAM-1 antibody was used to stain 5-microm paraffin sections. The slides were viewed at a magnification of x 40 by a blinded examiner. Twenty consecutive fields from standardly expanded tissue samples were viewed, and the volume density of PECAM-1 (V(V PECAM)) per parenchyma was measured, using point counting. In addition, 1-microm sections from 15 controls and 5 infants with CLD were stained with Toluidine blue and viewed under oil at a magnification of x 100, and the volume density of capillaries (V(V CAP)) and capillary load (CL) were calculated. The V(V PECAM) increased significantly with GA in controls (r = 0.72, P < 0.001). There was no relationship between V(V PECAM) and severity of CLD. Both V(V CAP) and CL increased significantly with GA (r = 0.93, P < 0.001; r = 0.94, P < 0.001, respectively). The infants with CLD had a normal or increased V(V CAP) and CL compared to controls. In summary, V(V PECAM), V(V CAP), and CL increased significantly with gestational age in control infants, but the postconceptional age range in CLD infants was too short to determine whether the V(V PECAM) changed. Infants with CLD had normal or increased V(V CAP) and CL compared to controls. The PECAM-1 immunostain does not appear to be a sensitive method for assessing capillary density in infants with CLD. These findings of normal or increased capillary load may represent a vascular adaptation for the lack of secondary septation and decreased surface area in CLD. PMID- 11921456 TI - Effects of tobramycin solution for inhalation on global ratings of quality of life in patients with cystic fibrosis and Pseudomonas aeruginosa infection. AB - In a previously published placebo-controlled trial, tobramycin solution for inhalation (TSI) was shown to improve lung function and other outcomes in patients with cystic fibrosis (CF). The objectives of the current study were to examine the effects of TSI on global ratings of health-related quality of life (HRQOL) by patients (or their parents) and physicians blind to group assignment, and to determine whether any perceived benefits persisted over time. The global ratings of HRQOL in 520 patients with CF and chronic Pseudomonas aeruginosa infection were analyzed retrospectively. Patients were randomly assigned to receive 24 weeks of placebo or treatment with TSI 300 mg b.i.d., both administered in cycles of 28 days on drug (or placebo) followed by 28 days off, for a total of three cycles. After each on-drug cycle, patients or parents, and physicians, were asked to rate whether the patient's condition was better, unchanged, or worse. There was strong agreement between the paired patient/parent and physician global HRQOL ratings across the three cycles. Regression analyses demonstrated that patients in the TSI group were significantly more likely to report improvements in HRQOL than were patients in the placebo group. This effect was found to be both immediate (end of on-drug cycle 1) and delayed (end of subsequent on-drug cycles 2 and 3) (P < 0.05). In addition, change in forced expired volume in 1 sec (FEV(1)) % predicted values was a significant predictor of improvement in HRQOL ratings by patients and parents. After controlling for change in FEV(1) % predicted, physician ratings showed significant improvement only at the end of cycle 1. Finally, controlling for initial lung disease severity, longitudinal growth models revealed that patients on TSI and their physicians reported higher HRQOL ratings than did placebo patients and their physicians across the three cycles; however, the magnitude of this effect decreased over time. Results of this study provided consistent evidence that TSI was associated with improved global ratings of HRQOL completed by both patients or parents, and physicians. Although these results are promising, they are limited by the use of a single-item rating of health. Future studies of the effects of TSI should utilize a well-validated, disease-specific measure of HRQOL. PMID- 11921455 TI - Improvements of lung function in cystic fibrosis. AB - Improved care for patients with cystic fibrosis (CF) has led to their improved survival. We analyzed retrospectively whether improvements in lung function (LF) could be detected in our CF patients over the decade 1980-1990. In 72 patients, 153 LF measurements were performed in their first year of life (1980-1991), and then 189 LF measurements were performed again in 60 of those patients during their sixth year of life (1987-1997). Regression analysis was performed on LF parameters at age 6 years. When adjusting for weight, height, gender, and LF in the first year of life, the date of subsequent measurement was positively associated with FEV(1) (P < 0.01) and MEF(50%) (P < 0.05) and negatively with FRC(pleth) (P < 0.05). The proposed model predicts a child's FEV(1) at age 6 to be 75% of predicted if born in 1980, but 108% of predicted when born in 1990. Improved CF care is the most likely explanation for this observation. PMID- 11921457 TI - Lung function in diastrophic dysplasia. AB - Diastrophic dysplasia (DD) is a rare skeletal dysplasia characterized by short limbed short stature, contractures and early degeneration of joints, and spinal deformities such as scoliosis. Mortality is increased in the neonatal period, in part due to tracheo- and bronchomalacia. Additionally, spinal deformities are very rigid, decreasing mobility of the chest cage. The aim of our study was to evaluate lung volumes and airway flow dynamics in patients with DD. A total of 31 patients (12 males, 19 females) underwent a detailed clinical examination measurements of standing height PA-radiography of the spine, flow-volume spirometry, and body plethysmography. The patients were assigned to two groups: children and adolescents (0-18 years, n = 18) and adults (over 18 years, n = 13). The mean spirometric parameters were mostly within the predicted value range, although the variation was wide. At least one abnormally low spirometry parameter was found in 6 (33%) of the children and adolescents and in 7 (54%) of the adults. Mean forced vital capacity (FVC) was 104% (range 48-163%) of predicted values in the children and adolescents and 95% (58-140%) of the adults. Peak expiratory flow (PEF) values were abnormal in 4 (22%) of the children and adolescents and in 5 (39%) adults. The mean plethysmographic parameters were all within the predicted value range. At least one abnormal plethysmographic value was found in 6 (33%) of the children and adolescents and in 4 (31%) of the adults. Airway resistance (Raw) was significantly higher in the adults than in the children and adolescents (P = 0.016), and was abnormally high in 3 (23%) of the adults. The angle of thoracic or thoracolumbar scoliosis correlated with the percentages of the predicted values of FVC (r(s) = - 0.66), forced expired volume in 1 sec (FEV(1)) (r(s) = - 0.56), and total lung capacity (TLC) (r(s) = - 0.67). Age correlated with the FEV(1)/FVC ratio (r(s) = - 0.41), with the maximal expired flow at 50% FEV (MEF(50)) values (r(s) = - 0.55), with the residual volume (RV) values (r(s) = - 0.47), and with the RV/TLC ratio (r(s) = - 0.43). Variable bronchial obstruction was found in 1 (6%) child and in 2 (17%) adults. Although the patients with DD had, on average, normal lung volumes, large individual variation occurred. Airway resistance was increased in adults. The angle of scoliosis correlated inversely with lung volume parameters. PMID- 11921458 TI - Exhaled nitric oxide concentrations: online versus offline values in healthy children. AB - Exhaled nitric oxide (FE(NO)) is a noninvasive and practical method to assess airway inflammation. We conducted this investigation to determine the most appropriate flow rate to measure FE(NO) and to obtain reference values for FE(NO) in children. FE(NO) was measured in 112 healthy 6-18 year olds (60 males) at 4 expiratory flow rates (46, 31, 23, and 15 mL/sec) using a chemiluminescent nitric oxide analyzer. Offline and online analyses were done to determine FE(NO) intraclass correlation coefficients, the relationship between FE(NO) and expiratory flow rates, and the effects of age and gender on these measurements. The major findings were: 1) intraclass correlation coefficients for FE(NO) and flow rates ranged from 0.92-0.99 for offline values, and 0.99 for all online values; 2) variation at an expiratory flow rate of 46 mL/sec (SD, 9.39) was considerably less than at other flows, especially at 15 mL/sec (SD, 26.55); 3) FE(NO) increased as flow rates decreased for both offline and online values; 4) there were no significant differences and good agreement between offline bag and online FE(NO) values at 31 and 46 mL/sec expiratory flows; and 5) using multiple regression, significant predictors of FE(NO) were flow, body surface area, age, and FEF(25-75). We have provided FE(NO) values in healthy children and propose that the ideal expiratory flow rate for FE(NO) measurements in children using the single breath technique is between 30-50 mL/sec. PMID- 11921459 TI - Mucociliary clearance in cystic fibrosis. AB - Abnormal mucociliary clearance (MCC) is one of the central hypotheses for the development of lung disease in patients with cystic fibrosis (CF). However, attempts to demonstrate this decrease of MCC in vivo have proved to be somewhat less definitive, with the evidence barely favoring impaired clearance. Any apparent disparities are most likely due to the variety of methodologies used by different laboratories to measure MCC. The limitations of the various methodologies are examined in this review, in an attempt to better facilitate comparison of results. A number of physical and pharmacological therapies have been developed to promote mucus clearance from the CF airway. A summary of the results of interventional studies utilizing the measurement of MCC as an outcome measure is presented. PMID- 11921460 TI - Irreversible airway obstruction in childhood asthma? A clinician's viewpoint. AB - Even though childhood asthma is assumed to comprise reversible airway obstruction, some children develop irreversible airway obstruction (not reversed by a bronchodilator or corticosteroids); this may be due to inflammation that has caused remodeling. Lately, it has been claimed that in the absence of treatment with inhaled corticosteroids, most patients will develop progressive irreversible obstruction. Several studies culminating with the Childhood Asthma Management Program (CAMP) study, which was the first randomized placebo-controlled prospective long-term study designed to test for irreversible obstruction, did not show the development of such progressive irreversible obstruction. Nevertheless, deterioration in pulmonary function does occur in some patients, probably due to inadequate anti-inflammatory treatment, and possibly also due to maintenance adrenergic treatment. Most previous studies concentrated on forced expiratory volume in 1 sec (FEV(1)), a test assessing mostly large airway obstruction. More studies are needed to investigate the presence of small airway obstruction. PMID- 11921461 TI - Hypersensitivity to inhaled TOBI following reaction to gentamicin. AB - Cystic fibrosis (CF) is the most common autosomal-recessive disease in Caucasians. Colonization with Pseudomonas aeruginosa (P. aeruginosa) of the CF airways causes deterioration of pulmonary status. TOBI (Tobramycin solution for inhalation) is an inhaled antibiotic that can improve the pulmonary disease. We report on a 9-year old boy with CF who developed a rash following a course of IV gentamicin. The rash resolved after its discontinuation. However, the rash returned all over his body, with the start of inhalation of TOBI therapy. We desensitized the patient using escalating doses of inhaled TOBI. He tolerated the procedure well, and continues to be on TOBI 9 months after desensitization on a once-a-day regimen. PMID- 11921462 TI - Normal volume of distribution of tobramycin in a mother and daughter with a CFTR splice mutation (1717 - 1G --> A). AB - A mother and daughter pair with CF who shared a splice mutation (1717 - 1G --> A) had a normal volume of distribution of tobramycin. The literature on tobramycin pharmacokinetics, which was published before the genetic defect was identified, is discussed. The authors speculate on the role of CFTR in the distribution of aminoglycosides and recommend that CFTR mutations should be clarified in all future studies of tobramycin pharmacokinetics in patients with CF. PMID- 11921463 TI - Primary bronchopulmonary leiomyosarcoma of the left main bronchus in a child presenting with wheezing and atelectasis of the left lung. AB - Primary lung tumors are rare in children, and primary bronchopulmonary leiomyosarcomas are very rare in children, with only 10 cases reported in the English-language literature. We report on the eleventh case of primary bronchopulmonary leiomyosarcoma in a child, and it is the first case in Taiwan, and also in Asia. In addition, this is the second case of this tumor arising in the left main bronchus. Before the present case, there were only 2 cases of this tumor in the main bronchus: one was in the right main-stem bronchus, and the other in the left-stem main bronchus. Ours is the second case showing pleural effusion. Chest roentgenogram and bronchoscopy are helpful in the diagnosis of primary bronchopulmonary tumors, but a definitive diagnosis can only be made by biopsy. The tumor is potentially curable if total excision is possible; irradiation has little therapeutic value, and early excision offers the only hope of cure. The child is alive and well 19 months after pneumonectomy, without having received radiotherapy or chemotherapy. PMID- 11921465 TI - [Directly etched bridges. Experiences in 33 patients]. AB - A study of immediately fabricated composite-etch bridges by using the crown of the extracted tooth as a pontic or by using a full-composite pontic is presented. The teeth to be replaced were to be extracted because of severe periodontal problems. The lifespan, the need for repair and possible advantages of this semi permanent bridgework are discussed. The results indicate a favorable prognosis for this type of treatment in cases where there is a wish for postponing the insertion of removable partial dentures or extensive crown and bridgework. PMID- 11921466 TI - [Hematology and dentistry. Part V. Clinical manifestations of hematological malignancies in the oral cavity]. AB - Haematological malignancies like acute and chronic leukemias, malignant lymphoma and plasma cell disorders may exhibit oral manifestation during their course. In this article the oral manifestation of a number of haematological malignancies presented in the oral cavity as well as the complications of the treatment are described. PMID- 11921467 TI - [Treatment for dental anxiety. Evaluation of treatment of patients with anxiety]. AB - This article presents the results of a study among 332 extremely anxious persons who applied for treatment at SBT, a Dutch special dental care clinic, between July 1990 and December 1991. It was found that 85% of these persons actually started treatment. Two and a half years later it appeared that 39% of them still received regular treatment, whereas 34% had completed treatment. Twelve percent stopped visiting before treatment was completed. Furthermore, results showed that dental anxiety was reduced after treatment. However, 36% of the patients still avoided appointments with a dentist. It is concluded that for most of the dentally anxious patients specialized care and oral health are no guarantee for regular dental attendance in the long term. PMID- 11921468 TI - [Information video about oral hygiene. Short term effects on knowledge, attitude and behavior of 1st and 2nd graders of LBO and MAVO]. AB - The short term effects of a dental health educational video on adolescents' knowledge, attitude and future behaviour were assessed. Results showed a large effect on knowledge and a small effect on five attitudinal aspects. No effects were found on future behaviour. PMID- 11921469 TI - [Oral function study. An electrical field between function and comfort]. PMID- 11921470 TI - [Oral function study. An electric field between function and physiology]. PMID- 11921471 TI - [A blue swelling on the tongue]. PMID- 11921473 TI - [Treating, negotiating and referring. Introduction]. PMID- 11921472 TI - [Dental caries caused by longterm use of antidepressive agents]. PMID- 11921474 TI - [Decision-making in dentistry]. AB - Decisions made by the general dental practitioner are not only based on technical considerations, but are also influenced by interpersonal factors. This might give an explanation for the fact that patients are treated in a variety of ways in otherwise comparable dental practices. This variety is more likely to be a sign of optimal individual health care, rather than of unacceptable differences. PMID- 11921476 TI - [Perspective of a patient in the general practitioners office and in the dental office]. AB - Both general practitioners and dentists acknowledge the importance of the patient's perspective and the demand for care, and, consequently, of good communication with their patients. In general practice, the concept of reason for encounter has proved to be very useful for gaining more insight in the nature and the importance of the patient's perspective. Data from the Amsterdam Transition project show that the general practitioner understands the patient's reasons for encounters very well, and that the nature of the patient's reasons for encounter clearly affect the subsequent interventions. In this article, 260 letters concerning people's experiences with their dentist are used in order to provide an impression of communication problems in dentist practice in the Netherlands. Three major problem areas are identified, with a total of ten subcategories. Several of these are well known to the general practitioner as well. In addition, some problems are characteristic for the dentist practice. It is suggested to incorporate the concept of reason for encounter in dental care as an essential part of the description and analysis of the communication between dentists and their patients. PMID- 11921475 TI - [Defensive behavior in medical care. A lesson for the dentist?]. AB - Defensive behaviour in medical care is defined as a clear deviation from the physicians (dentists) usual behaviour and from what he/she considers to be good practice in order to prevent problems in the contact with patients and their families. General practitioners behave defensively in substantial proportion of their referrals to a specialist (15%) and of their ordering diagnostic tests (25%). In both instances varying (clinical and non-clinical) reasons can prompt the physician to refer or to test. Defensive considerations can play an equally varying role within all these reasons, which shed more light on the nature of the daily work in primary care. It is concluded that no reason exists to presume that Dutch dentists are more free either from non-clinical considerations and from defensive behaviour when they treat their patients than general practitioners. PMID- 11921477 TI - [Defensive behavior? Also in dentistry?]. AB - The objective of this pilot-study was to investigate if defensive behaviour of dental practitioners occurs just as in the general practice of family physicians. On the basis of a survey, developed in the Department of General Practice of the University of Amsterdam, 38 dentists (mainly general-practitioners with an average of 20.9 years in practice), were interviewed. The dentists mention defensive practices, but the reasons are unrelated to fear of law-suits. They recommend unnecessary treatments, based on the wishes of their patients. The financial situation of the patient and defensive behaviour are closely related. The respondents avoid to give treatments because of dentally unmotivated patients or deviant behaviour of their visitors. Unnecessary referrals to specialists are also cited. Defensive behaviour was found to play a role in 3.5% of all patient encounters. PMID- 11921478 TI - [Second opinion in dental practice. The use of a dental information point (TIP)]. AB - The term 'second opinion' is a well-known phenomenon in dentistry. In the Netherlands, this phenomenon has mainly been worked out in five so called Dental Information Points (TIP). The way in which these offices function and the kind of problems they have to cope with have been described in this article. Finally, recommendations are given to enable the general practitioner to provide the patient with a well-considered second opinion. PMID- 11921479 TI - [Treating, negotiating and referring. Perspective of the dentist and specialist]. AB - Dentists and dental specialists are in general well aware of the professional treatment standards. Nevertheless, it is occasionally necessary or unavoidable to adjust or even to ignore these standards in the management of an individual patient. PMID- 11921480 TI - [Harmony of the facial profile. Part III. Vertical rules]. AB - Rules for profile harmony in the vertical direction reflect the acceptable relations between the heights of the different facial parts. The relation between the maxillary and nasal heights determines the acceptable range for the mandibular height. PMID- 11921481 TI - [Hyperthyroidism and dental treatment]. AB - In this contribution the cause, consequences, and treatment of hyperthyroidism are discussed. The consequences in relation to dental treatment are given special attention. A patient with a properly treated hyperthyroidism can safely undergo dental treatment, although periods of stress are to be avoided. PMID- 11921482 TI - [A perfect set of teeth!]. PMID- 11921483 TI - [Orthodontics and growth?]. PMID- 11921484 TI - [Craniomandibular dysfunction?]. PMID- 11921485 TI - [Crohn disease, a mysterious disorder]. PMID- 11921486 TI - Joint Commission zeroes in on disaster plans: is yours up to par? AB - The Joint Commission says that surveyors are paying close attention to disaster planning. You'll need to demonstrate compliance with many new standards, including performing a hazard vulnerability analysis and working with community organizations. All ED staff members must be prepared to answer questions about their role in disaster planning. Include individuals from community organizations when developing your disaster plan and holding drills. PMID- 11921487 TI - Nine ways to use handheld computers. AB - Handheld computers can be used to make calculations, share articles with colleagues, store contact information, and even save lives. Translation software can facilitate communication with non-English-speaking patients. You can find out the cost of medications with drug reference software. Make the ED schedules accessible, so that staff can volunteer to fill in gaps. PMID- 11921488 TI - CMS says you'll get paid for observation. PMID- 11921489 TI - Bioterrorism watch. Ease of access to deadly chemicals may be the greatest threat to hospital readiness. PMID- 11921490 TI - Beta-lactam/beta-lactamase inhibitor combinations in empiric management of pediatric infections. AB - Beta-lactam antibiotics have long played a central role in the management of pediatric infections. However, widespread beta-lactam resistance among community- and hospital-acquired pathogens, mainly due to beta-lactamase production, has reduced the usefulness of these trusted and well-tolerated agents. Many regions have reported an increase in beta-lactamase-mediated resistance to cephalosporins and carbapenems as well as penicillins among clinically important Gram-positive and Gram-negative aerobes and anaerobes. For some pathogens such as Moraxella catarrhalis, Klebsiella species and Pseudomonas aeruginosa, virtually all strains worldwide are beta-lactamase producers. The development of beta-lactamase inhibitors for co-administration with a number of established beta-lactam agents has restored their usefulness in pediatric patients. The combination of ampicillin plus sulbactam has broad anti-aerobic and anti-anaerobic activity in vitro and achieves high concentrations in many body tissues and fluids. The availability of a mutual oral prodrug, sultamicillin, has enabled the development of an oral formulation. Excellent clinical response and bacterial eradication rates with ampicillin/sulbactam and sultamicillin have been demonstrated for upper and lower respiratory tract infections, urinary tract infections, osteomyelitis, and meningitis in pediatric patients and neonates. Furthermore, many studies have demonstrated an excellent tolerability profile. Thus, ampicillin/sulbactam has an important role in the management of pediatric infections. PMID- 11921491 TI - Experience with ampicillin/sulbactam in severe infections. AB - The emergence of beta-lactamase-mediated resistance to established beta-lactam antibiotics prompted the development of beta-lactamase inhibitors for co administration. Ampicillin has been combined with sulbactam for both parenteral and oral (as the mutual pro-drug sultamicillin) administration. The combination is active in vitro against a wide variety of Gram-positive and Gram-negative pathogens, including aerobic and anaerobic organisms. In clinical trials, ampicillin/sulbactam has proved clinically and bacteriologically effective against a variety of frequently encountered pediatric infections, including mild to-moderate upper respiratory tract infections (acute otitis media, sinusitis, pharyngitis, and tonsillitis), severe post-operative and intra-abdominal infections, periorbital infections (which, left untreated, can lead to blindness, brain abscess, or death), acute epiglottitis, bacterial meningitis, and brain abscess. Ampicillin/sulbactam has also proved effective in the prevention of post operative surgical infections in pediatric patients. The clinical efficacy profile of ampicillin/sulbactam and sultamicillin, combined with their excellent tolerability profile, make these agents attractive options for the management of many life-threatening infections in pediatric patients. PMID- 11921492 TI - Beta-lactam resistance: clinical implications for pediatric patients. AB - The emergence of resistance to established antibiotic agents such as beta-lactams has been reported worldwide and poses a serious challenge to the management of pediatric infections. The most common mechanism of resistance involves the production of an enzyme that inactivates the antibiotic before it can be effective. Streptococcus pneumoniae, the most common cause of pediatric respiratory tract infections, exhibits variable resistance to penicillins and aminopenicillin due to alterations in its penicillin-binding proteins (PBPs). Haemophilus influenzae and Moraxella catarrhalis show moderate and high beta lactamase-mediated resistance to aminopenicillins, although they remain susceptible to beta-lactam/beta-lactamase inhibitor combinations. Methicillin resistant Staphylococcus aureus, a frequent cause of skin and soft-tissue infections, has shown PBP-mediated beta-lactam resistance, prompting the wide spread use of vancomycin to eradicate this pathogen. Finally, PBP-mediated resistance has been observed in a large proportion of isolates of coagulase negative staphylococci, which account for a high proportion of nosocomial infections, particularly in neonatal intensive care units. The challenge is to control the emergence of beta-lactamase-mediated resistance by using beta-lactams judiciously. In this regard, the beta-lactam/beta-lactamase inhibitor combinations have an important role to play in extending the usefulness of established beta-lactam agents. PMID- 11921493 TI - A multicentre study of abdominal aorta diameters in a Turkish population. AB - The aim of this study was to determine the normal values for aortic diameters and the prevalence of aortic dilatation in a mixed Turkish population. Between March 1998 and May 2000, patients who were undergoing abdominal ultrasonography examination for pathologies not involving the aorta, in three different cities, were enrolled into the study prospectively. The anterior posterior aortic diameters were measured at the subdiaphragmatic and aortic bifurcation levels using ultrasonography. A total of 596 patients were included (302 females, 294 males). The mean age was 48 +/- 16 years (range, 6-88 years). The mean aortic diameter in the whole group was 19.0 +/- 3.9 mm (10-45 mm) at the subdiaphragmatic level and 15.7 +/- 3.6 mm (9-65 mm) at the aortic bifurcation level. The mean subdiaphragmatic aortic diameter was 18 +/- 3 mm in females and 19 +/- 4 mm in males. The mean aortic diameters at the bifurcation level was 15 +/- 3 mm in females and 16 +/- 4 mm in males. An aortic bifurcation diameter > 30 mm was encountered in 0.67% of the population. This ratio increased to 1.8% in patients over 55 years of age, regardless of sex. A subdiaphragmatic aorta diameter above 30 mm was observed in 1.2% of the population. In patients over the age of 55 years, this ratio increased to 2.7% (3.6% in males and 1.9% in females). In this national study, the subdiaphragmatic aortic diameters were similar to mean values reported in the world literature. The mean aortic bifurcation diameters were generally lower when compared with the literature, which may be due to difficulties in standardization of the measurements. Aneurysmal dilatation rates in this study also conform to those reported in studies conducted in other countries. Considering the significant number of patients with aneurysmal dilatation of the aorta in the elderly population, we believe it would be prudent to evaluate the aorta in all patients undergoing abdominal ultrasonographic examination. PMID- 11921494 TI - The effects of valsartan on insulin sensitivity in patients with primary hypertension. AB - Insulin resistance is an important risk factor of cardiovascular disease. This study was performed to determine the effects of valsartan on insulin sensitivity in patients with primary hypertension. In this study, non-obese subjects with primary hypertension and a reference group of healthy subjects matched by age, sex and body mass index were evaluated; patients with any other causes of peripheral insulin resistance and hyperlipidaemia were excluded. The effect of valsartan on insulin resistance, assessed by homeostasis model assessment (HOMA IR), fasting serum insulin levels, determined by radioimmunoassay, and fasting blood glucose concentrations, measured by the glucose oxidase method, were evaluated. All obtained data were evaluated by Pearson correlation analysis. Before valsartan treatment, the fasting serum insulin levels were significantly elevated in the 20 hypertensive patients with primary hypertension compared with the 20 subjects in the reference group (19.6 +/- 7.1 versus 8.7 +/- 1.9 microIU/ml). The fasting serum insulin levels correlated with HOMA-IR. Correlation analysis also showed a significant relationship between HOMA-IR and both systolic and diastolic blood pressures (r = 0.71 and r = 0.77, respectively). In our study, we showed that patients with primary hypertension have a decreased insulin sensitivity that was reflected in high serum fasting insulin levels. Anti-hypertensive treatment with valsartan increases insulin sensitivity. PMID- 11921495 TI - Effects of fluvastatin treatment on insulin sensitivity in patients with hyperlipidaemia. AB - This study aimed to determine the effects of fluvastatin treatment on insulin sensitivity in patients with hyperlipidaemia. Non-obese, normoglycaemic, normotensive patients with hyperlipidaemia (n = 20) and a reference group of healthy subjects of similar age, sex, and body mass index (n = 20) were evaluated. Patients with other causes of peripheral insulin resistance were excluded. All participants underwent a diagnostic protocol, which included measurements of insulin sensitivity index and other metabolic parameters. Insulin sensitivity was assessed by Homeostasis Model Assessment (HOMA). Serum insulin levels were tested by radioimmunoassay. Patients were treated with fluvastatin 40 mg once daily for 3 months. Before fluvastatin treatment, fasting serum insulin levels were significantly raised in patients with hyperlipidaemia compared with subjects from the reference group (19.1 +/- 13.4 versus 8.1 +/- 3.4 microIU/ml). The fasting serum insulin levels and HOMA-estimated insulin sensitivity were correlated in the whole group. Correlation analysis showed a significant relationship between HOMA-estimated insulin resistance and plasma cholesterol and triglyceride concentrations. Patients with hyperlipidaemia had reduced insulin sensitivity that was reflected by high serum fasting insulin levels. Anti hyperlipidaemic treatment with fluvastatin increases insulin sensitivity. PMID- 11921496 TI - The impact of acute dynamic exercise on intraocular pressure: role of the beta 2 adrenergic receptor polymorphism. AB - Effects of mutations in the beta 2-adrenergic receptor (beta 2AR) gene on intraocular pressure (IOP), in response to acute dynamic exercise, were investigated in 19 healthy males (age 22.6 +/- 2.8 years). Intraocular pressures were measured pre- and post-exercise. Weight, height, body mass index, and maximal oxygen (VO2max) uptake were recorded and subjects were genotyped for Arg16Gly, Gln27Glu and Thr164Ile mutations of the beta 2AR gene. Post-exercise, reductions in mean IOP values were found in 16 subjects with the Gly16Gly and Arg16Gly genotypes, but these values remained low in the eight patients with the Gly16Gly genotype 3 h post-exercise, whereas they returned to baseline within 1 h in the eight subjects with the Arg16Gly genotype. beta 2AR stimulation during exercise could be an important regulator of IOP response and determining beta 2AR polymorphisms may improve understanding of pathogenesis and treatment selection in ophthalmic diseases, e.g. glaucoma. PMID- 11921497 TI - Effects of the concomitant administration of tamsulosin (0.8 mg/day) on the pharmacokinetic and safety profile of theophylline (5 mg/kg): a placebo controlled evaluation. AB - We performed a single-blind, sequential-design study to investigate the effects of concomitant oral tamsulosin 0.8 mg/day on the pharmacokinetics and safety of intravenous theophylline 5 mg/kg in healthy subjects. Ten healthy volunteers aged 19-39 years received placebo on study days 0, 1, 2 and 10 and tamsulosin on days 3-9. Theophylline was administered intravenously on days 1 and 9. Theophylline and tamsulosin pharmacokinetic data were determined following administration of the drugs on days 1 and 9 and day 9, respectively. No differences were observed in theophylline pharmacokinetic parameters with and without concomitant tamsulosin, and there were no abnormalities in tamsulosin pharmacokinetic data. Some significant changes in vital signs and a number of mild adverse reactions were reported, but the overall safety profile of tamsulosin and theophylline was acceptable. The results of the study suggest that no dose adjustment in tamsulosin is necessary when it is administered concomitantly with theophylline. PMID- 11921498 TI - Efficacy of metadoxine in the management of acute alcohol intoxication. AB - This randomized, open-label study evaluated the efficacy of 300 mg metadoxine (given intravenously) added to standard treatment compared with standard treatment alone in managing the physical and psychological signs of acute alcohol intoxication. Fifty-two acutely intoxicated patients were randomly assigned to one of two groups and followed during a 2-h period. Changes in clinical symptoms, degree of intoxication, and blood alcohol level were monitored. More patients receiving metadoxine in addition to standard therapy significantly improved by at least one degree of intoxication (one clinical category) compared with those receiving standard treatment alone (76.9% versus 42.3%, respectively). Metadoxine treated patients also exhibited a significantly greater decrease in blood alcohol concentration compared with those receiving standard treatment alone (-105.4 +/- 61.5 mg/dl versus -60.1 +/- 38.6 mg/dl, respectively). Metadoxine improved the clinical signs of acute alcohol intoxication and accelerated alcohol clearance from the blood, thus supporting existing data. In contrast to previous data, these effects were concurrent but independent. No adverse effects were observed with metadoxine therapy. PMID- 11921500 TI - Clinical correlates of prolonged pain in Japanese patients with acute herpes zoster. AB - To determine which risk factors are relevant to the occurrence of post-herpetic neuralgia in Japanese patients with acute herpes zoster, correlations between the prolongation of pain and various disease factors were examined in 263 adult patients presenting within 10 days of the onset of rash at 17 institutions in the Hyogo region of Japan. All patients in whom pain persisted for more than 3 months were over 60 years of age. The pain also tended to be more prolonged in those with clustered vesicles, disturbed sleep and hypanaesthesia. Other factors such as underlying disease states, critically involved regions, scar tissue, generalized rash and allodynia were not relevant to the duration of pain. Although decreased pain persistence was observed in patients in whom acyclovir therapy was initiated within 72 h of the onset of symptoms in comparison with those in whom it was initiated after this time, the difference between the two groups of patients was not statistically significant. Our results suggest that advanced age, the presence of clustered vesicles, and disturbed sleep and hypanaesthesia influence the prolongation of herpes zoster pain. PMID- 11921499 TI - Plasma thrombospondin in immune thrombocytopenic purpura. AB - Patients with immune thrombocytopenic purpura (ITP) rarely suffer life threatening haemorrhages despite significant thrombocytopenia, probably because large numbers of hyperfunctioning platelets are present. Thrombospondin is a platelet alpha-granule protein and its plasma level may reflect platelet activation. We assessed circulating thrombospondin levels in 12 newly diagnosed ITP patients (one man; 11 women, aged 36 +/- 16 years) before they were treated for ITP. Twelve healthy people (four men; eight women, aged 31 +/- 11 years) acted as controls. Plasma thrombospondin concentrations were measured using enzyme-linked immunoassays. Thrombospondin concentrations tended to be higher, despite thrombocytopenia, in ITP patients (158.8 +/- 28.2 ng/ml) compared with controls (120.7 +/- 18.2 ng/ml). The difference was not statistically significant, but the relatively high circulating thrombospondin concentrations we observed suggest that residual platelets could be activated in ITP, thus indicating a more benign clinical course compared with aplastic thrombocytopenia. PMID- 11921501 TI - Hydatid disease of the liver in children: evaluation of surgical treatment. AB - The medical records of 55 paediatric patients with hepatic hydatidosis, who were treated between 1990 and 2001 at Ataturk University, Turkey, were reviewed retrospectively. The most common symptoms at presentation were abdominal mass (32.7% of cases) and pain (81.8% of cases) in the right upper quadrant of the abdomen. Cysts were found in the right lobe in 41 patients, in the left lobe in four patients and in both lobes in 10 patients. Multiple hepatic cysts were present in 12 cases and eight patients also had cysts in other organs. Surgical procedures were evacuation of the cyst and management of the cavity with tube drainage, capitonnage, omentoplasty, cystectomy or segmentectomy. Long-lasting biliary fistula (two patients) and cholangitis (two patients) developed following evacuation and tube drainage, and one patient developed cholangitis after capitonnage. This review suggests that omentoplasty and capitonnage are more effective than tube drainage in the management of the cyst cavity. PMID- 11921502 TI - Patient compliance and persistence with anti-hyperglycemic therapy: evaluation of a population of type 2 diabetic patients. AB - The persistence and compliance of type 2 diabetic patients to different regimens of anti-hyperglycemic therapy were assessed retrospectively. The pharmacy claims from a pharmacy benefit management organization were analysed from the third quarter of 1996 to the fourth quarter of 1999. Of the 23,400 patients enrolled and initiating anti-diabetic therapy, 85% started treatment with monotherapy, 9.5% with insulin alone, 4.1% with polytherapy and 1.3% with insulin plus another therapy. Monotherapy patients were characterized as receiving metformin, sulfonylurea or another agent. For the 1-year follow-up period, 70.5% of the metformin patients, 75.3% of the sulfonylurea patients and 86.8% of the polytherapy patients underwent no regimen modification (except discontinuation). For the patients who had no modification of their medication regimen, persistence with sulfonylurea or metformin monotherapy was 65% greater than with polytherapy over a 1-year period. Compliance with sulfonylurea or metformin monotherapy was 45% greater than with polytherapy. PMID- 11921503 TI - Thyroid abnormalities in lithium-treated patients with bipolar affective disorder. AB - The thyroid functions of 42 subjects with bipolar affective disorder receiving regular lithium therapy were analysed and their thyroid glands were examined by ultrasonography. Following the receipt of lithium therapy (duration 4-156 months), three subjects displayed subclinical hypothyroidism (7.1%), three subclinical hyperthyroidism (7.1%) and one hyperthyroidism (2.4%). Moreover, goitre was detected in 16 (38.1%) subjects. An increase in the conversion of free thyroxine (T4) to free tri-iodothyrosine (T3), which is an indication of mild thyroid dysfunction, was identified in 20 (47.6%) subjects, and was mostly seen in male subjects under 40 years of age and in those having weight gain. In conclusion, some thyroid dysfunctions were observed in the patients treated with lithium. PMID- 11921504 TI - Serum E-selectin and beta 2-microglobulin levels in Behcet's disease. AB - Behcet's disease is a systemic vasculitis of unknown aetiology. Endothelial cell injury plays an important role in the pathogenesis and immunopathology of systemic vasculitises, but an immunopathogenic basis is also probable in Behcet's disease. E-selectin is an indicator of endothelial injury, and beta 2 microglobulin (beta 2M) is increased in immunological disorders. The serum concentrations of these two markers were determined in 40 patients with active Behcet's disease and 40 healthy controls. E-selectin was determined by enzyme linked immunosorbent assay and beta 2M was determined by the nephelometric method. The mean (+/- SD) serum concentration of E-selectin was 122.13 +/- 62.32 ng/ml and that of beta 2M was 0.31 +/- 0.05 mg/dl. Both concentrations were statistically significantly increased in patients with Behcet's disease compared with controls. More comprehensive studies are needed to determine whether these parameters are useful indicators of the activity of Behcet's disease. PMID- 11921506 TI - The effects of cardiopulmonary bypass on androgen hormones in coronary artery bypass surgery. AB - The effects of testosterone on coronary vasomotor regulation have been described by several recent reports. Here we investigated changes in serum androgen levels during and after cardiopulmonary bypass (CPB) in patients who had undergone coronary artery bypass surgery. Serum luteinizing hormone, free testosterone and dihydroepiandrestenedione sulphate (DHEA sulphate) levels were evaluated in 38 male coronary artery bypass surgery patients using a chemical immunoassay technique. All hormone levels were corrected to account for haemodilution. Serum free testosterone level decreased significantly during weaning from CPB (from 15.7 +/- 4.2 nmol/l to 6.2 +/- 2.8 nmol/l), and an even greater decrease was observed in the first post-operative day (5.4 +/- 3.1 nmol/l). On the seventh post-operative day, free testosterone levels reached a normal value (11.8 +/- 5.5 nmol/l), although they were still significantly lower compared with the pre operative value. There were slight alterations in serum DHEA sulphate levels, although the only significant decrease occurred from the first to the seventh day post-operation (from 4.7 +/- 2.2 mumol/l to 3.7 +/- 1.8 mumol/l, respectively). Serum luteinizing hormone levels were decreased during weaning from CPB (from 4.8 +/- 2.1 mIU/ml to 3.9 +/- 1.8 mIU/ml), but increased rapidly to the pre-operative value (5.5 +/- 2.5 mIU/ml) at the first post-operative day. These results show that CPB affects serum luteinizing hormone, free testosterone and dihydroepiandrestenedione sulphate levels. The free testosterone level decreases significantly both during and after CPB surgery. PMID- 11921507 TI - [A nurse in Germany]. PMID- 11921505 TI - Atrioventricular nodal re-entrant tachycardia with two functionally discrete fast pathways. AB - We present a case with two forms of atrioventricular nodal re-entrant tachycardia (AVNRT) that revealed similar H-A-V sequences, but could be differentiated only by their retrograde atrial activation sequences. Both tachycardias were induced following anterograde slow pathway conduction, suggesting the slow pathway as the anterograde limb of the re-entry circuit. The earliest atrial activation site of one form was in the same region of the bundle of His as that of the common type of AVNRT, while that of the other form was the ostium of the coronary sinus. Properly timed extra-stimuli delivered from the atrium or ventricle during the latter tachycardia penetrated through the fast pathway without resetting the tachycardia cycle length. These rare phenomena suggest the existence of two functionally discrete fast pathways, of which the alternative pathway alters to become the more predominant retrograde limb according to time and circumstances. PMID- 11921508 TI - [Care of burns]. PMID- 11921509 TI - [Surgical care of burns in children]. PMID- 11921510 TI - [Tissue banks]. PMID- 11921511 TI - [Cardiovascular protection in women: benefits, how far?]. PMID- 11921512 TI - [Education of the cardiac patient]. PMID- 11921513 TI - [The patient or the disease?...]. PMID- 11921514 TI - [Neonatal jaundice]. PMID- 11921515 TI - [Draft of legislation relating to the rights of the patient and the quality of the health system]. PMID- 11921516 TI - [Sildenafil. A chemical solution to the (physical) breakdown of love]. PMID- 11921518 TI - [Reducing the consumption of salt, a stake in public health]. PMID- 11921517 TI - [Insulin injection. Insulin, its mode of action, injection sites, the material]. PMID- 11921519 TI - [Burns and home care]. PMID- 11921520 TI - [Blockage of tariffs, formation, 35 hours...the syringe is full]. PMID- 11921521 TI - Malaria control: achievements, problems and strategies. AB - Even if history has not always been the Magistra vitae, Cicero expected it to be, it should provide, as Baas said, a mirror in which to observe and compare the past and present in order to draw therefrom well-grounded conclusions for the future. Based on this belief, this paper aims to provide an overview of the foundations and development of malaria control policies during the XX century. It presents an analysis of the conflicting tendencies which shaped the development of these policies and which appear to have oscillated between calls for frontal attack in an all-out campaign and calls for sustainable gains, even if slow. It discusses the various approaches to the control of malaria, their achievements and their limitations, not only to serve as a background to understand better the foundations of current policies, but also to prevent that simplistic generalisations may again lead to exaggerated expectations and disillusion. The first part of the paper is devoted to the development of malaria control during the first half of the century, characterised by the ups and downs in the reliance on mosquito control as the control measure applicable everywhere. The proliferation of "man-made-malaria", which accompanied the push for economic development in most of the endemic countries, spurred the need for control interventions and, while great successes were obtained in many specific projects, the general campaigns proposed by the enthusiasts of vector control faced increasing difficulties in their practical implementation in the field. Important events, which may be considered representative of this period are, on the campaign approach, the success of Gorgas in the Panama Canal, but also the failure of the Mian Mir project in India; while on the developmental approach, the Italian and Dutch schools of malariology, the Tennessee Valley and the development of malaria sanitation, included the so called species sanitation. The projection of these developments to a global scale was steered by the Malaria Commission of the League of Nations and greatly supported by the Rockefeller Foundation. Perhaps the most important contribution of this period was the development of malaria epidemiology, including the study of the genesis of epidemics and their possible forecasting and prevention. Although the great effectiveness of DDT was perhaps the main determinant for proposing the global eradication of the disease in the 1950s, it was the confidence in the epidemiological knowledge and the prestige of malariology, which gave credibility to the proposal at the political level. The second part deals with the global malaria eradication campaign of the 1950s and 1960s. It recognises the enormous impact of the eradication effort in the consolidation of the control successes of the first half of the century, as well as its influence in the development of planning of health programmes. Nevertheless, it also stresses the negative influence that the failure to achieve its utopian expectations had on the general disappointment and slow progress of malaria control, which characterised the last third of the century. The paper then analyses the evolution of malaria control funding, which often appears out of tune with political statements. The fourth part is devoted to the search for realistic approaches to malaria control, leading to the adoption of the global malaria control strategy in Amsterdam in 1992, and the challenge, at the end of the century, to rally forces commensurate with the magnitude of the problem, while aiming at realistic objectives. After discussing the conflicting views on the relations between malaria and socio economic development and the desirable integration of malaria control into sustainable development, the paper ends with some considerations on the perspectives of malaria control, as seen by the author in early 1998, just before the launching of the current Roll Back Malaria initiative by WHO. PMID- 11921522 TI - [Diabetic nephropathy: is it possible to prevent it or to delay its progression?]. PMID- 11921523 TI - [Does cholestasis change the clinical usefulness of CA 19-9 in pacreatobiliary cancer?]. AB - BACKGROUND: CA 19-9 is used for diagnosis of gastrointestinal neoplasia, mainly pancreatic and biliary cancer. False positive results have been described in cholestasis. OBJECTIVE: To establish the clinical value of CA 19-9 in the diagnosis of pancreatic and biliary cancer in patients with and without cholestasis. METHODS: Five hundred forty-eight medical records of patients with serum CA 19-9 determination performed from May-1996 to June-1998 were reviewed. Cases were grouped by final diagnosis; malignancy was established by histology or clinical and radiological characteristics. ROC curves were used to calculate ideal cut-off values (ICV) for the test. Cholestasis was defined as bilirrubinemia above 3 mg/dL. RESULTS: Thirty percent of serum determinations were done in patients with non-pancreatic and non-hepatobiliary benign diseases (only 1.3% with values > or = 100 U/mL). CA 19-9 levels were higher in pancreatic and hepatobiliary malignancy compared to benign diseases of the same origin, as well as in pancreatic cancer when compared with hepatobiliary cancer. ICV for differentiation of malignant hepatobiliary diseases was set around 100 U/mL, with increased specificity when compared with the usual cut-off value (37 U/mL). Cholestasis increased the values of the antigen in malignant and benign diseases and modified the efficacy of the test by increasing sensitivity while decreasing specificity. The ICV for determining resectability in pancreatic tumors was 224 U/mL. CONCLUSIONS: CA 19-9 is a valuable test for diagnosis of malignant pancreato-hepatobiliary disease. Given that cholestasis modifies the operational characteristics of the test, a cut-off value has to be tailored for each patient depending on the clinical setting, so to maintain the usefulness of the marker. PMID- 11921524 TI - Comparing easy and accessible parameters of glycemic control in type 2 diabetes. AB - BACKGROUND: HbA1c is considered the gold standard of long-term glycemic control and is recommended as a routine test for every diabetic patient. However, its common use in clinical practice has some problems related to lack of standardization and its relative cost. Recent studies have suggested, that postprandial blood glucose could be better than a fasting sample, as a marker of diabetes control. The objective of the present study was to evaluate the relative value of plasma glucose samples at different times of the day, and easy and accessible programs for home blood and urinary glucose measurements compared with HbA1c in assessing the mean glycemic control of type 2 diabetic patients. METHODS: Sixty type 2 diabetic patients were instructed to do home blood and urine glucose monitoring for two months, at the end, plasma glucose profiles were obtained. RESULTS: The mean of all the capillary BG measurements had the best correlation with the HbA1c (r = 0.84, p < 0.001), followed by the mean of the capillary BG measurements before breakfast and supper (r = 0.82, p < 0.001), and the 2 hr. postbreakfast plasma glucose (r = 0.79 p < 0.001). The fasting PG had a low correlation (r = 0.65, p < 0.001), but a good sensitivity to predict a fair or a poor metabolic control. Diabetes duration and type of treatment explained 17% and 28% of variance in HbA1c levels. CONCLUSIONS: A bimonthly fasting PG correlated well with the glycosylated hemoglobin and is the easiest and cheapest way of monitoring glycemic control in type 2 diabetic patients with some preserved insulin reserve (diabetes for less than 10 years and on treatment with only one hypoglycemic agent). A sample of capillary BG, fasting, once per week correlates better with the HbA1c than a fasting PG every 2-3 months. The 2 hr and 5 hr postbreakfast PG have a good correlation with the HbA1c, but are not a substitute for doing BG monitoring. Glycosuria may be a useful parameter to rule out a fair or poor metabolic control in some patients. PMID- 11921525 TI - Characterization of rpoB gene mutations in rifampicin resistant Mycobacterium tuberculosis strains isolated from pulmonary tuberculosis patients at 5 Mexican public hospitals. AB - OBJECTIVE: To characterize the rpoB gene mutations of the rifampicin-resistant M. tuberculosis strains isolated in pulmonary tuberculosis patients from Mexico. MATERIAL AND METHODS: Thirty-seven clinical M. tuberculosis isolates cultured on Lowenstein-Jensen media and obtained from consecutive tuberculosis patients in 5 public hospitals were analyzed by PCR and the INNO-LiPA Rif TB for amplification and detection of mutations associated with rifampicin resistance, respectively. RESULTS: Twenty-three out of 37 isolates (62.2%) were found to be wild type (rifampicin susceptible), while 14 isolates (37.8%) contained mutations associated with rifampicin resistance. Seven out of the 37 isolates (18.9%) had a delta S1 mutation, in the nucleotide position number 511; one (2.7%) had a R4b mutation, in nucleotide H526D; five (13.5%) contained a R5 mutation, in nucleotide S531L; and one (2.7%) showed a double mutation delta S1/R4b. CONCLUSION: According to the marker used (rifampicin resistance), at least five different strains of M. tuberculosis circulate among pulmonary tuberculosis patients in Mexico. rpoB gene mutations associated with rifampicin resistance are common in Mexico. A single mutation in nucleotide 511 was the most frequently observed, followed by single mutations in nucleotides S531L and H526D. PMID- 11921526 TI - Correction of insulin resistance in methimazole-treated patients with Graves disease. AB - The effect of thyroid hormone excess on glucose tolerance as well as insulin secretion and its peripheral action has been a matter of debate for many years. Thyrotoxicosis caused by Graves' disease is associated in some patients with impaired glucose tolerance and insulin resistance. The objective of the present study was to investigate if the insulin sensitivity, assessed by the euglycemic insulin clamp technique, increase after correction of hyperthyroidism due to Graves' disease. After four months of medical treatment, patients became euthyroid and insulin sensitivity increased significantly from 3.47 to 6.39 mg/kg/min; therefore it was concluded that insulin resistance could be improved after successful treatment of hyperthyroidism. The precise mechanism underlying the effect of thyroid hormone excess on insulin sensitivity remains to be elucidated. PMID- 11921527 TI - [Epidemiologic features of tetanus in adults form Yucatan, Mexico]. AB - OBJECTIVE: To describe epidemiologic features from tetanus in adult patients (TIAP) treated at hospital general O'Horan in Merida, Yucatan, Mexico and compare them with another Mexican series analyzed 25 years ago. MATERIAL AND METHODS: From 1985 to 1999, 121 TIAP cases aged 13 years or older were identified from which 112 were analyzed. Diagnosis of TIAP was made just in a clinical basis. To analyze information inferencial statistics were used. RESULTS: People affected by tetanus averaged 43 +/- 21, 95% CI 39 to 47. A 3:1 male to female ratio was documented. Eighty two (73%) patients come from the rural area; 43 (38%) did work as peasants. Sixty two cases (55%) were diagnosed during the Fall and Winter seasons. In 91 patients (81%) no anti-tetanus vaccination was documented. In 89 cases (79%) incubation period averaged 5.4 +/- 4 days, 95% CI 5 to 6. According to this 89 cases (79%) with incubation period < 10 days were graded as severe tetanus and 23 (21%) with incubation period > or = 10 days were graded as non severe tetanus. Tetanus-prone wounds were documented in 95 (85%) cases, 59 (62%) of which (62%) were localized in the lower extremities. Final outcome dichotomized either as death patient (group one) or surviving patient (group two) was documented in 103 cases of whom 67 (65%) were in group one and 36 (35%) were in group two. By comparing them, differences were seen in mean age (P = 0.004, 95% CI 3.9 to 19.8), age categories (< 50 vs. > or = 50) (chi 2 P = 0.001, 95% CI 0.6 to 0.60), severity of tetanus (Fisher exact test P = 0.0009, 95% CI 2 to 53) and mean hospitalization time (mean difference 14.8, P = 0.0001, 95% CI 11 to 18) but not in sex (chi 2 0.69, P = 0.40). CONCLUSIONS: In the State of Yucatan, Peninsula de Yucatan, Mexico, TIAP is still an endemic process with high mortality rate specially among young productive people. Secondary preventive measures as routinely tetanus toxoid booster vaccination are still not enough, at least in adulthood. PMID- 11921528 TI - [Endovascular treatment of arterial occlusive disease of the iliac arteries]. AB - Atherosclerotic disease is one of the most important health problems in the United States and in the entire world. The endovascular techniques for the treatment of peripheral vascular disease have been investigated in recent years. This is a review of the literature of the endovascular techniques used in the treatment of occlusive iliac disease. Angioplasty has showed excellent results in concentric lesions, of less 5 cm of length with a technical success of 96%, primary patency of 90%, 81% and 72% at one, two and three years. The complication rate is 0.5% to 0.8% and the mortality rate is 0.2%. The use of stents has improved the success of arterial recanalization, with a technical success of 97%, and a primary patency of 90%, 84% and 71% at one, two and three years. Major complications appear in less than 1% of the cases. In conclusion the endovascular techniques have been proved to be the treatment of choice for certain selected forms of presentation of atherosclerotic disease in the iliac arteries, specifically short, focal lesions. The results have been excellent with less mortality and morbidity than surgery. PMID- 11921529 TI - [Treatment of Hodgkin's lymphoma and hematopoietic cell autotransplantation]. PMID- 11921530 TI - [Pathogenesis of hepatitis C virus infection]. AB - The hepatitis C virus (HCV) is the leading cause of chronic liver disease worldwide. Chronic hepatitis C is a mayor cause of cirrhosis and hepatocellular carcinoma and HCV-related end-stage liver disease is, in many countries, the first cause of liver transplantation. HCV infection is characterized by its propensity to chronicity. Because of its high genetic variability, HCV has the capability to escape the immune response of the host. HCV is not directly cytopathic and liver lesions are mainly related to immune-mediated mechanisms that are characterized by a predominant type 1 helper cell response. Co-factor influencing the outcome of the disease including age, gender and alcohol consumption are poorly understood and other factors such as immunologic and genetic factors may play and important role. Recent studies have shown that the combination therapy with alpha interferon and ribavirin induces a sustained virological response in about 40% of patients with chronic hepatitis C. The lack of animal models and of in vitro cultures systems hampers the understanding of the pathogenesis of chronic hepatitis C and the development of new antivirals. The conjugation of polyethyleneglycol improved the pharmacodynamics and the efficacy of alpha interferon. The development of an effective vaccine remains the most difficult challenge. Because of the high protein variability of HCV, protective vaccines could be extremely difficult to produce and therapeutic vaccines seem more realistic. Considerable progress has been made in the field of HCV since its discovery 10 years ago but a major effort needs to be made in the next decade to control HCV-related disease. PMID- 11921531 TI - [Molecular physiology of salt absorption in the loop of Henle]. PMID- 11921532 TI - [Pearl S. Buck, Literature Nobel Price, and phenylketonuria: a moving relationship]. AB - Art and Medicine, in some occasions have singular contact points. An example of this is the life of the North American writer Pearl S. Buck, a Litterature Nobel Prize winner in 1938, and her relation with phenylketonuria and mental retardation. The present paper is a biographical sketch of this brilliant writer, whose only daughter had phenylketonuria, an inborn error of metabolism of low frequency. If this disease is not treated, it may cause mental retardation. At the same time, we point out some of the highlights of the discovery of this disorder in Norway in 1934, and the description of its first treatment in 1953. At the present time, the mental retardation that this disease causes, can be prevented by means of the neonatal screening. PMID- 11921533 TI - [Repetitive transcranial magnetic stimulation as adjuvant in antidepressive pharmacologic treatment]. PMID- 11921534 TI - [HLA genes and the origin of Amerindians]. PMID- 11921535 TI - A nodular pulmonary lesion due to Linguatula serrata in an HIV-positive man. AB - A coin-shaped pulmonary lesion was accidentally detected in a 42-year-old, HIV seropositive man residing in Bari (Apulia, Southern Italy) during a routine X-ray examination. A lung cancer was suspected, obliging physicians to investigate surgically. After thoracotomy a lung nodule, 1.8 cm in diameter, was excised and submitted for histological examination. Histological analysis revealed a nodular infarctual lesion containing a larva of Pentastomida. Despite the poor state of preservation of the parasite it was possible to recognise some morphological characteristics which enabled the parasite to be identified as Linguatula serrata (Pentastomida, Porocephalida). This is the first case reported in Europe in the lung in a living man due to this parasite, the few others occurring in autopsy reports. No evident correlations were found in the present case between HIV seropositivity and the development of the parasitosis. The importance of lung nodules caused by metazoan invertebrates is emphasised: even though they are rare in man, they are regularly mistaken for cancer at X-ray examination. PMID- 11921536 TI - Comparative study of anti-Hypoderma antibody kinetics in sera, single and bulk milk samples of naturally infested cattle. AB - The investigation was carried out in Basilicata region (Southern Italy) from October 1997 to June 1998. Fifteen dairy cows bred in semiconfined conditions on a farm with a history of hypodermosis were sampled once a month for sera and milk; bulk milk from these animals was also collected monthly from the farm's tanker. Samples were tested for anti-Hypoderma spp. antibodies (Abs) with an ELISA technique and clinical parasitological examination was carried out monthly from January to July on all the animals in order to detect grubs. Blood and single and bulk milk samples yielded similar antibody kinetics and patterns in accordance with results obtained in previous immunological surveys in Italy. All animals were warbled in the spring time. November-January was confirmed to be the most suitable period for seroepidemiological survey for weather conditions in Southern Italy. The ELISA test proved once again to be very useful because it is simple to perform and cost effective. Either blood or milk samples may be used for epidemiological surveys; bulk milk may be very useful for the preliminary detection of hypodermosis on farms or in areas where there is no data available on the diffusion of the disease. PMID- 11921537 TI - [Persistent scalp infestation by Dermanyssus gallinae in an Emilian country woman]. AB - A case of persistent infestation of the scalp due to Dermanyssus gallinae.- Infestation due to Dermanyssus gallinae, the common red mite of poultry, in a country-woman aged 69 years from Crevalcore (Emilia-Romagna region, Northern Italy), is described. The case was unusual either for its location, the scalp, and for its persistence, 9 months. Specimens of the mite were also found in the henhouse adjacent to the patient's house. Apparently the woman recovered by means of daily washing of camomile tea. PMID- 11921538 TI - [Fungistatic activity of a sesquiterpene lactone (tomentosin) isolated from fresh Inula viscosa (Asteraceae) flowers from the Puglia region]. AB - A sesquiterpene lactone, tomentosin, has been isolated and identified form Inula viscosa Ait (Asteraceae) flowers. The molecule, at 1 mg/ml concentration, showed to be active in vitro against Microsporum canis, Microsporum gypseum and Trichophyton mentagrophytes. PMID- 11921540 TI - Goat warble fly infestation by Przhevalskiana silenus (Diptera: Oestridae): immunoepidemiologic survey in the Basilicata region (southern Italy). AB - The demonstration of serological cross-reactivity between the Hypoderma lineatum antigen and anti-Przhevalskiana silenus antibodies led us to prepare an immunological test (ELISA) for an early diagnosis of goat warble fly infestation. Using the Hypodermosis ELISA-Kit (Vetoquinol Diagnostic, France) produced for the immunodiagnosis of bovine hypodermosis, an epidemiological assay was carried out in Basilicata region where goat breeding is very common and no data are reported with regards to the distribution of goat warble fly infestation. Out of a total of 1,100 flocks and 41,200 goats, 105 randomly extracted flocks proved to be infected and 262 sera out of 1,316 were positive; goat warble fly infestation proved to be present in Basilicata with values similar to those recorded in the surrounding regions (Apulia and Calabria). Statistical evaluation showed highly significant differences between the number of infected flocks in the mountainous areas and hills and those of the mountainous areas and the plain, but no differences between hills and plains. The higher number of positive sera and antibody titres in November-December confirmed that these months are the optimal period for sampling sera in order to perform an early diagnosis. The ELISA test was confirmed to be an easy and economic tool especially when sera sampled within a brucellosis eradication program are used. PMID- 11921539 TI - Gastro-intestinal nematode infections in four Alpine chamois herds: influence of host density on helminth egg output. AB - A three years survey on gastro-intestinal nematode egg output was carried out in four Alpine chamois herds (Rupicapra rupicapra rupicapra) from Belluno province (Italy). Data were analysed in order to detect the influence of cattle presence and chamois density on egg output, and to evaluate the stability of host-parasite relationship. As expected, variations related to the season were present, with the major peak of egg output observed in the summer. Results highlighted an unexpected inverse influence of host density on egg counts. The results are in relation to host-parasite relationship stability. Parasites seem to be able to self-regulate in the absence of parasite-induced host mortality. No effect due to cattle presence was detected. PMID- 11921541 TI - [Dientamoeba fragilis and dientamoebiasis: aspects of clinical parasitology and laboratory diagnosis]. AB - The flagellate Dientamoeba fragilis is a potential intestinal pathogen of humans. It is probably undervalued overall because its identification relies on permanent stains directly on fresh fecal specimens (Giemsa) or preserved in PVA or SAF (Trichrome or Iron-hematoxylin). During 1999 we processed stoll samples of 151 subjects and we searched also D. fragilis. We processed 105 cases of enteritis (82 children and 23 adults) and 46 samples of formed stools to investigate only for intestinal parasites (12 children and 34 adults). The microscopical observations were made on smears directly in saline solution, and after Dobell solution, MIF and Giemsa stained. Iron hematoxylin and trichrome stains were performed in some circumstances only. D. fragilis was observed in 11.3% of cases, 7.6% among the group of enteritis and 13.8% among the other subjects. These positive 17 cases were observed more often among adults (76.5% of cases) than among children (23.5%). In our study area D. fragilis seems more frequent than Giardia intestinalis and represents the third important intestinal parasite, following Enterobius vermicularis (children) and Strongyloides stercoralis (elders). PMID- 11921543 TI - Molecular identification of Borrelia valaisiana and HGE-like Ehrlichia in Ixodes ricinus ticks sampled in north-eastern Italy: first report in Veneto region. AB - PCR amplification was applied to screen the presence of both Borrelia burgdorferi s.l. and Ehrlichia species in pools of field-collected Ixodes ricinus ticks. The specimens so far analysed (n = 55), grouped in 11 pools, were sampled in Feltre area (Veneto region, NE Italy). Five pools proved positive for B. valaisiana (45%) and one of them (9%) was also positive for Ehrlichia, that was further characterised as a HGE-like Ehrlichia. This is the first report of the two bacteria in the Veneto region. The pool positive for both pathogens was used to adjust a multiplex PCR assay, which allowed the detection and identification of both parasites in a single experiment. The advantages offered by this assay, when standardised, will substantially broaden the perspectives of ecological and epidemiological investigations on animal/human Lyme disease and ehrlichiosis, greatly facilitating disease surveillance and control programs. PMID- 11921542 TI - Efficacy of moxidectin for the prevention of adult heartworm (Dirofilaria immitis) infection in dogs. AB - The authors report the efficacy of orally administered moxidectin for the prevention of canine heartworm infection in two endemic areas in northern Italy. Two trials were conducted on a total of 257 dogs, including 137 treated with moxidectin (minimum dose of 3 mcg/kg body weight), 85 with ivermectin (minimum dose 6.6 mcg/kg b.w.) and 35 untreated controls. Results of testing for microfilariae and circulating adult female antigens were negative for all treated dogs at the end of both trials. No adverse reactions to moxidectin were observed. In the study areas, prevalence values for Dirofilaria immitis infection calculated on the basis of the untreated controls and testing dogs which had no preventive treatment in the previous transmission season ranged 23-65%. This study confirms the efficacy and safety of moxidectin in the prevention of adult heartworm infection in dogs. PMID- 11921544 TI - Cryptosporidium sp. in a free ranging house gecko (Hemidactylus turcicus) in Israel. AB - A house gecko, Hemidactylus turcicus, from an Israeli locality was found infected by Cryptosporidium sp. Infection was recovered in the anterior gut, upon examination by a transmission electron microscope. It comprised from 2.9 x 3.3 microns undivided and 2.6-3.3 x 3.8-3.9 microns dividing meronts, 1.9-2.3 x 2.0 2.6 microns young macrogamonts, and a 3.3 x 3.9 microns sporulated oocyst containing in cross section 4 sporozoites and a residuum. Oocyst dimensions were smaller and apparently not conspecific with the Cryptosporidium sp. reported from the introduced H. turcicus in Texas. PMID- 11921545 TI - Third report of ocular parastrongyliasis (angiostrongyliasis) from Sri Lanka. AB - A further case of ocular parastrongyliasis has been seen in a patient from Sri Lanka. As it is a juvenile female worm it could not be identified to the species, but it is probably Parastrongylus (= Angiostrongylus) cantonensis which is the commonly reported species in the country. This is the third authentic case of such an infection in Sri Lanka in recent years. PMID- 11921546 TI - [Chromosomal polymorphism of populations of Anopheles arabiensis (Diptera: Culicidae) from Reunion island and cross-fertility among continental African populations]. AB - Cytological examination of a sample of Anopheles gambiae complex mosquitoes from Reunion island revealed the presence of An. arabiensis only. Chromosomal polymorphisms were observed only for inversion 3Ra, the standard homozygote form being predominant. Cross-mating experiments with laboratory specimens originating from continental Africa produced viable and fertile offspring with no chromosomal asynapsis observed in the F1 female progeny. There was no evidence for speciation of the Reunion island populations. The results are discussed with regard to the behaviour of the vector and its influence on the vectorial capacity of this species, and the history of malaria and malaria control in the South-West islands of the Indian Ocean and on Reunion island in particular. PMID- 11921547 TI - Ethiopian Medical Journal: looking into the future. PMID- 11921548 TI - Anatomical variations of gallbladder and biliary ducts among Ethiopians. AB - The extrahepatic biliary system was investigated to determine the types and frequency of variations in a group of Ethiopian subjects. A total of 110 specimens were obtained from adult Ethiopian subjects. Each specimen consisting of the whole of the liver, stomach, duodenum and the lesser omentum was removed and dissected carefully to expose the gallbladder and biliary ducts without disturbing their relationship. The majority (51%) of subjects had supramarginal type of gallbladder. The neck and infundibulum of the gallbladder were shorter in the present group compared to reports in other populations. There were significantly more cases of kinking of the gallbladder and Hartmann's pouch in the female than in the male subjects. However, there was no significant difference between the present and previously reported groups as well as the male and female subjects of the present group in the length or course of the cystic, hepatic or bile ducts. The findings in the present study suggest that variations in the gallbladder and the biliary ducts seen in the Ethiopian group are similar to what is reported in other populations. The significantly higher prevalence of kinking of the gallbladder and Hartmann's pouch in the female than in the male subjects may have some contribution to the higher rate of gallstone formation and biliary tract diseases in females. PMID- 11921549 TI - Cryptococcosis in patients from Tikur Anbessa Hospital, Addis Ababa, Ethiopia. AB - A study was done to investigate the occurrence of Cryptococcus neoformans infection in patients admitted to Tikur Anbessa Hospital. Cryptococcus neoformans is an important opportunistic fungal pathogen in immunocompromised patients. The study was done over a period of 18 months, from October 1998 to April 2000. During this period, a total of 1088 cerebrospinal fluid (CSF) specimens were sent to the bacteriology laboratory in Tikur Anbessa Hospital, out of which 275 were subjected for India ink examination. Out of these 19 (7%) were India ink positive. Additionally one lymph node aspirate was also India ink positive. All these specimens were culture positive for Cryptococcus neoformans. The medical records of these patients were retrospectively reviewed and all presenting clinical symptoms were recorded. In this group of patients with meningitis the most common presenting were fever and headache. In addition, at the time of admission, 75% of the patients had other opportunistic infections, such as oral candidiasis, herpes zoster and Pneumocystis carini pneumonia. The mortality rate was high even in patients with appropriate therapy. All the isolates were identified as C. neoformans var. neoformans. The variety gatti was not isolated in this study. PMID- 11921550 TI - Trends of acute myocardial infarction admissions over a decade, Tikur Anbessa Hospital. AB - This is a case series analysis undertaken to evaluate the importance of acute myocardial infarction (AMI) as a cause of admission at the Tikur Anbassa medical intensive care unit (MICU) and the trend of that importance over a decade (1988 1997). Clinical presentations and the frequencies of major coronary risk factors in the individual patient were also assessed for the later half of the decade (1993-1997). In the decade under study 2313 patients were admitted to MICU according to its register. Overall AMI was the third commonest cause of admission and accounted for 8.8% (N = 203) of all MICU cases. AMI annual admissions increased consistently over the years. Of the 122 AMI admissions during the second half of the decade, 92 charts were available for detailed analysis. 86% (79/92) fulfilled the stated criteria for the diagnosis. The mean age of these patients was 55.1 +/- 13.0 years. Males constituted 82% of all AMI cases. Eighty seven percent (69/79) of the cases were first admissions with the diagnosis of AMI. Ninety-four percent (74/79) of them were brought to the emergency room due to chest discomfort and 20% were in frank pulmonary oedema. Nineteen percent died in hospital. 81% and 34% of the patients had one or more and two or more major coronary risk factors excluding age and gender respectively. Raised total cholesterol and hypertension were commonest risk factors being reported in 69% and 47% of AMI patients respectively. In conclusion, this study has demonstrated that AMI is indeed on the rise at least at Tikur Anbassa Teaching Hospital, and possibly at other health institutions. The conventional coronary risk factors seem to operate in the Ethiopian series as well. The study highlights the need for coronary risk factors surveys at least in the susceptible population group to assess the gravity of the problem. PMID- 11921551 TI - Clinical description of children with HIV/AIDS admitted at a referral hospital in Addis Ababa. AB - Seventy-seven definitely HIV infected children were identified at Ethio-Swedish Children's Hospital in Addis Ababa, Ethiopia, between January 1990 and April 2000. There were 39 females and 38 males. Their age ranged from 15 months to 12 years and their median age was 3.8 years. Most commonly diagnosed diseases included disseminated tuberculosis seen in 28 patients, pneumonia in 27, pulmonary tuberculosis in 19, persistent or chronic diarrhoeal disease in 21, otitis media in 17 and marasmus in 17 patients. Pneumocystic carinii pneumonia was considered in 11 patients. One patient was suspected to have lymphoid interstitial pneumonitis. Sixteen children (21%) died in hospital. There was an abandoned child and 20 children were already orphans. All patients presented with clinical opportunistic infections and thus there is a need to develop a pediatric clinical guideline for patient management. Primary prevention of HIV infection in women of the child bearing age is of paramount importance and reduction of mother to-child transmission of HIV infection by antiretroviral drug needs to be considered as a primary prevention for a child. PMID- 11921552 TI - The use of morbidity questionnaires to identify communities with high prevalence of geohelminth infections in Gondar region, Ethiopia. AB - In this preliminary study, a total of 8006 schoolchildren (aged 9-19 years) from 77 elementary schools was screened for infection with Ascaris lumbriocoides, Trichuris trichiura and hookworms using the Kato-Kata method. A. lumbriocoides was commonly encountered (35.4%) followed by the hookworms (16.3%) and T. trichiura (12.7%). The prevalence of "intestinal worms" (infection by A. lumbricoides, T. tirchiura and/or hookworms) accounted to 50.6%. The children were also asked about their recent experiences of the following: 'diarrhea', 'stomach ache', 'blood in stool', 'distended stomach'. and 'worms' using questionnaires and gave 24.0%, 64.9%, 18.6%. 37.0%, 38.1% 'yes' answers, respectively. At individual level reported 'worms' correlated with the prevalence of A. lumbricoides [OR 7.39 (95% CI: 6.65-8.20)] and 'intestinal worms' [OR 4.72 (95% CI: 4.27-5.22)]. Reported 'diarrhoea' was the best answer for T. trichiura [OR: 4.58 (95% CI: 3.98-5.27)]. Among schools, there were similar associations between prorated 'worms' and the prevalence of A. lumbricoides [OR 0.63 (95% CI: 0.47-0.75)] and 'intestinal worms' [OR 0.55 (95% CI: 0.37-0.69)]. No sign or symptom was significantly associated with hookworm infection. Reported 'worms' had the best though moderate diagnostic performance for 'intestinal worms' (sensitivity: 82.6%, specificity 51.6%, positive predictive value: 66.7%, negative predictive value: 71.7%) and A. lumbrocoides (sensitivity: 91.8%, specificity: 50.0%, positive predictive value: 75.5%, negative predictive value: 64.3%). These results indicate the potential of using reported 'worms' as a means of rapid assessment for identifying communities with a high prevalence of geohelminth infection. PMID- 11921553 TI - A survey of statistical methodology used in Ethiopian health science research journals. AB - Results from many research efforts have been generated from the use of statistical methods. However, most researchers use data analysis as the only component of statistics to arrive at their results. There is a concern that this alone may not yield the appropriate result if it is not done with due understanding of and regard for study design, data acquisition techniques, choice of sample, and methods of statistical analysis. This paper attempts to document how much of these statistical methods are in use in Ethiopian health science research journals. All the original articles, in the two health science research journals--Ethiopian Medical Journal and The Ethiopian Journal of Health Development published between 1995 and 1999 were surveyed. A total of 232 papers were evaluated to see how far their authors have complied to these basic requirements as well as the statistical software used. The results of the survey demonstrate that in about 80% of the papers, the study design has been specified, 50% employed cross-sectional or survey designs, 14% provided detailed information on how sample size was determined and of this group 37% employed probabilistic selection methods. About 84% of the papers did not mention clearly what statistical methods they intended to employ to answer their research questions. Compared to others, attaching variability to a statistic using +/- SD or SE, t statistics and P values were more frequently misused. Only 57% used computers to manage their data and do statistical analysis. EPI-INFO (a statistical software for Epidemiology) was used in 61% among the users of computer software. Considering the important roles of health science journals in guiding and updating good medical practice, low level and inappropriate use of statistical methodologies in the surveyed journals should give cause for concern. It is, therefore, recommended that a series of continuing education in statistics is done periodically to enhance the knowledge of health science researchers as well as editors and peer reviewers of health science journals to expand their background in statistical methods and acquaint them with new techniques. PMID- 11921554 TI - Neonatal tetanus in Awassa: retrospective analysis of patients admitted over 5 years. AB - Neonatal tetanus is a significant health problem worldwide, with an estimated 500,000 deaths per year. Retrospective analysis of medical records of neonates admitted to Bushulo Major Health Center with the diagnosis of tetanus neonatorum over 5 years period (1994/98) was made to describe the clinical profile and management out come. There were a total of 43 neonates admitted with a diagnosis of tetanus during the study period. Male to Female ratio was 2:1. Mean age at presentation (incubation period) was 9.5 days (range; 4-20 days). Most patients were brought within 3 days of the onset of symptoms. Common presenting complaints were; rigidity, spasm, failure to suck, trismus, fever and seizure. The overall case fatality was 29/42 (69%). All patients with severe classification and 23/32 (72%) of patients with moderate severity died. Most deaths occurred during the first two days of hospitalization (20/29 died within 2 days), and survivors stayed in hospital for a long period (8-26 days). Improved immunization coverage, clean delivery and clean cord care are recommended to reduce morbidity and mortality of neonatal tetanus. PMID- 11921555 TI - Comparative study of adult polycystic kidney disease. AB - An Ethiopian series dealt with 4 cases of adult polycystic kidney disease and mentioned the paucity of it in analyses of hospital admission in African literature. The present series on the lgbos of Nigeria, West Africa, compares well with the Ethiopian experience and constitutes one more group to be placed on the world map of this intriguing disease. PMID- 11921556 TI - Juvenile nasopharyngeal angiofibroma presenting as Foster Kennedy Syndrome. AB - Juvenile nasopharyngral angiofibroma (JNA) is a rare benign tumor of the nasopharynx that occurs in adolescent boys with epistaxis and nasal obstruction. It may grow into the cranium causing elevated intra-cranial pressure and compression on the optic nerve. A histological-proven case of JNA in an 18 year old Ethiopian boys is presented. He became blind due to optic atrophy in the right eye, but salvaged a useful vision in the left eye radiotherapy. The possibility of Foster-Kennedy Syndrome, a presentation of one atrophic and one papilloedematous optic nerve head of bilateral asymmetric optic atrophy, is discussed. Controversies about its histological appearance, natural history, diagnostic methods and management modalities are reviewed. JNA should be considered in adolescent boys who present with optic atrophy and/or nasal mass. Early detection and initial surgical treatment with adjunct radiotherapy could have prevented visual loss on this boy. PMID- 11921557 TI - The HIV pandemic and surgery. PMID- 11921558 TI - HIV infection in general surgical patients at the Ga-Rankuwa/MEDUNSA complex South Africa. AB - OBJECTIVE: To assess the possible impact of HIV infection on the management of general surgical patients at the Ga-Rankuwa Hospital. DESIGN: A prospective study. SETTING: Ga-Rankuwa Hospital/Medical University of Southern Africa (MEDUNSA) Academic Complex, Pretoria, South Africa. SUBJECTS: Nine hundred and forty one patients admitted to general surgical wards. MAIN OUTCOME MEASURES: HIV infection and CD4 counts. RESULTS: Nine hundred and forty one patients admitted to general surgical wards from January 1966 to December 1997 were tested for HIV infection. Twelve per cent tested positive. HIV positive patients were significantly younger [33 +/- 10 versus 41 +/- 7 (men +/- SD) years, Chi-square = 51, p < 0.0001]. There was no correlation of HIV positivity to the patient's sex (p = 0.7). In forty three HIV positive patients treated surgically, mortality was attributed to HIV/AIDS in only one patient with a CD4 count of 47/ul who died following laparotomy for peritonitis. CD4 counts done during follow up in thirty one HIV positive patients revealed a count of < 500/ul in eleven patients. CONCLUSION: It is predicted that an increasing number of patients with HIV/AIDS will be admitted to general surgery wards of the Ga-Rankuwa Hospital. Surgeons are advised to take universal precautions to prevent HIV infection. PMID- 11921559 TI - Oral manifestations of HIV/AIDS in a Kenyan provincial hospital. AB - BACKGROUND: In Kenya many patients exposed to the HIV infection present with orofacial lesions as the primary manifestations of the disease and only a few studies have been performed to document this observation. OBJECTIVE: To clinically evaluate and document the range and pattern of oral lesions in a group of hospitalised patients with HIV-infection. DESIGN: A prospective study. SETTING: Coast Province General Hospital in Mombasa, Kenya, which is the main referral institution serving a population of approximately two million people. METHODS: Examination of all the cases included in the study was performed according to the WHO criteria. Both male and female patients aged 16 years and above were selected. The criterion of recruitment was based on a suspicion of immunosuppression, the presence of oral manifestations and the willingness to participate in the study. Prior to the examination each patient had undergone counselling followed by two consecutive screening tests using the ELISA technique. Where indicated incisional biopsy was performed to confirm the clinical diagnosis of the relevant lesions. In collaboration with the medical team, treatment was administered as per the needs of the patient in terms of anti fungals, antivirals or topical cortisteroids. RESULTS: Of the 61 cases, 25(41%) were males and 36(59%) females with an age range of 19 to 65 years (mean = 34.7 years). While all the cases had periodontal disease, over 80% had candidiasis of the hyperplastic, erythematous and pseudomembraneous types. Lymphadenopathy and angular cheilitis were each diagnosed in 27.9% of the cases; while oral Kaposi's sarcoma was seen in 13% of the patients. Other conditions seen included persistent oral ulceration (11.5%), oral hairy leukopLakia and herpes zoster each constituting 4.9%; herpes simplex, mucosal hyperpigmentation, parotomegaly and facial palsy each comprised six per cent and oral warts seen in one case. In accordance with the pattern and prevalence of oral manifestations in our study, the results were largely consistent with those documented elsewhere. CONCLUSION: For the alleviation of the morbidity arising from the commonly occurring lesions, early detection is mandatory. Furthermore, documentation of the varied regional patterns of occurrence of these lesions may aid in the rational application of the emerging treatments. PMID- 11921560 TI - Malignant lymphoma in western Ethiopia. AB - OBJECTIVE: To identify the pattern of malignant lymphoma in western Ethiopia and compare it with similar studies conducted in the region. DESIGN: A retrospective study. METHODS: A search was made for all histopathologic diagnosis of malignant lymphoma from the files of the department for the years 1988-1999. The pathologic diagnosis of Hodgkin's disease was grouped according to Rye's classification and non-Hodgkin's lymphoma according to the working formulation for clinical usage classification. The results were tabulated and analysed. SETTING: The department of pathology of Gondar College of Medical Sciences; the only unit which provides histopathologic services for the region, that includes Amhara and Tigrai region. SUBJECTS: All lymphoma cases diagnosed at the department were included. RESULTS: A total of 83 cases of lymphoma were diagnosed over a period of 11 years (1988 1999). Their age ranged from four years to 79 years. Twenty one (25.3%) had Hodgkin's diseases, 61(73.5%) non Hodgkin's lymphomas and one (1.2%) unspecified lymphoma. The commonest type of Hodgkin's disease were lymphocyte predominant (38%) and mixed cellularity (33.3%). Nodular sclerosing Hodgkin's disease was diagnosed in only one case. The commonest type of non-Hodgkin's lymphoma was high grade lymphoma (41.0%) followed by low grade lymphoma (32.8%). Burkitt's lymphoma was diagnosed in four cases. The frequency of Hodgkin's disease was high in the first and second decades while non-Hodgkin's lymphoma showed high frequency in the fifth decade. CONCLUSION: This study has shown many similarities in proportion of lymphomas and age and sex distribution of cases. The identified minor differences such as frequencies of histologic patterns of Hodgkin's disease and paucity of Burkitt's lymphoma deserve explanation and invites prospective studies. PMID- 11921561 TI - Tonsillectomy and adenotonsillectomy in Sudanese patients. AB - OBJECTIVES: To highlight the indications for and complications of tonsillectomy or adenotonsillectomy operations in Sudanese patients and to find out whether any correlation can be deduced between routinely requested pre-operative investigations and the complications which may arise. DESIGN: A prospective study. SETTING: Khartoum ENT Teaching Hospital, Khartoum, Sudan. SUBJECTS: One hundred and twenty patients, 55% females and 45% males, age range 3 to 50 years. Pre-operative investigations consisted of a complete blood count and bleeding and clotting times., ASO titres and urinalysis. INTERVENTIONS: Consisted of tonsillectomy or adenotonsillectomy performed on in-patients by one of the authors under general anaesthesia. MAIN OUTCOME MEASURES: High erythrocyte sedimentation rate (ESR) or ASO titres should not deter an indicated tonsillectomy operation which can still be carried out if meticulous haemostasis is observed. RESULTS: Three or more episodes of exudative tonsillitis per year and upper airway obstruction were the commonest indications for tonsillectomy or adenotonsillectomy accounting for 72.5% and 16.7% of the operations. Intra operative bleeding occurred in 63 (52.5%) of the patients. Low figures of reactionary and secondary haemorrhages were seen. CONCLUSION: It is concluded that as bleeding, both intra- and post-operative is the main complication of tonsillectomy, its incidence can be reduced to minimal levels by careful selection of patients, in whom the operation is carried out under general anaesthesia with gentle handling of tissues and detention of the patients post operatively for two to three days. We also found out that coagulation tests need not be routinely requested and reserved for patients with a history of bleeding tendencies. PMID- 11921562 TI - Role of fine needle aspiration biopsy in the management of thyroid nodules. AB - OBJECTIVE: To evaluate the degree of utilisation, the accuracy and the effect of fine needle aspiration biopsy (FNAB) in patients with nodular thyroid lesions. DESIGN: Cross-sectional study. SETTING: King Khalid University Hospital, Riyadh, Saudi Arabia. SUBJECTS: Four hundred and twenty eight patients with thyroid lesions who were treated surgically were reviewed. The FNAB of 201 patients were compared with the post-operative histopathological findings. RESULTS: Most of the patients (69%) presented with nodular thyroid disease (solitary and multinodular) and among this group 31.4% did not undergo FNAB pre-operatively. In more than 76%, the FNAB resulted in a definite cytologic diagnosis. A sensitivity of 71.4% and a specificity of 96.4% was achieved. CONCLUSION: If FNAB were used routinely in the initial evaluation of patients with thyroid nodules it could have an enormous effect on their management. It reduces the number of patients going for thyroidectomy and eliminates the need for nuclear scans to screen patients with thyroid nodules. Therefore, we recommend FNAB to be part of the initial evaluation of all patients with nodular thyroid disease. PMID- 11921563 TI - Survival of Fuji IX ART fillings in permanent teeth of primary school children in Tanzania. AB - OBJECTIVE: To evaluate the clinical performance of atraumatic restorative treatment (ART) fillings using Fuji IX as a filling material in field conditions. DESIGN: Longitudinal study of the ART fillings in permanent teeth of primary school children aged eight to fifteen years. SETTING: Primary schools in Morogoro municipality, Tanzania. SUBJECTS AND METHODS: Standard 3 and 4 children in five primary schools randomly selected from a list of 36 primary schools of Morogoro municipality were examined for dental caries and periodontal conditions. All 296 carious lesions that were indicated for restoration were treated using ART approach according to the instructions given in the manual for ART approach for the control of dental caries. Essential measurements for treated teeth and cavity were taken. The cavities were filled with Fuji IX glass ionomer cement as per manufacturer's instructions. After one year, 238 restorations were evaluated using the criteria for evaluating ART restorations. MAIN OUTCOME MEASURE: Clinical appearance of the surface of the restorations. RESULTS: Ninety four per cent of the restorations evaluated were rated as good and intact, while 1.7% were rated as having slight defects that needed no repair, giving a one year survival rate of 96.1%. Mean working time was 14.5 minutes. CONCLUSIONS AND RECOMMENDATIONS: The one-year survival rate of 96.1% is high enough to recommend wide use of ART in Tanzania. Town and municipal councils should be encouraged to adopt ART in their school oral health programmes. PMID- 11921564 TI - Female sterilisation through mini-laparotomy at Gondar College of Medical Sciences. AB - OBJECTIVES: To describe the socio-demographic characteristics and clinical course and follow up of clients who have undergone voluntary surgical contraception (VSC) through minilaparotomy (mini-lap) under local anaesthesia (LA). Also, to evaluate the safety and method satisfaction, so as to forward recommendations for method utilisation in the Ethiopian context. DESIGN: A case series design where pre- and post-operative conditions of clients coming for voluntary surgical contraception via mini-laparotomy were systematically analysed. SETTING: Department of Obstetrics and Gynaecology, Gondar College of Medical Sciences, Gondar, Ethiopia. SUBJECTS AND METHODS: Eighty two clients (median age of 33 years, range 25-40) who decided to use tubal sterilisation method of contraception from April 1993 to May 1995 were included in the study. A format prepared and distributed to VSC providing sites in Ethiopia by the Family Guidance Association of Ethiopia (FGAE) was used in collecting the necessary information, including informed consent of every client. MAIN OUTCOME MEASURES: Previous knowledge of contraceptive methods, decision making for tubal sterilisation, size of incision, advantages of use of local anaesthesia in the local setting, duration of hospital stay, conditions on follow up. RESULTS: Eighty two (55.4%) women underwent tubal sterilisation through mini-lap. The mean number of the live children per client was 6.2 +/- 1.7, with parity ranging from two to eleven children. Mothers with five or more children were 70 (85.4%). Among 69 mothers (84.1%), the last pregnancy outcome were live births. The average length of the time since making a decision not to have any more children was 2.5 +/- 2 years (median of one year). The rationales given were economic, ill health and completed family size. Regarding the pattern of decision making, in 77 (91.9%) clients both the couples were involved in decision making. All except two underwent the procedure under LA and no complication was encountered during surgery. Follow-up attendance was 100% in one-year period and nothing serious was reported. All reported to be satisfied with the method. CONCLUSION: The study showed that early decision making by involving both couples other than proper case selection minimises regrets. The authors believe that tubal sterilisation through mini-lap under LA is an ideal method in developing countries where access to family planning and other reproductive health services are not widely available and where there is population explosion, less than 10% contraceptive prevalence rate and high maternal mortality. PMID- 11921565 TI - Birthweight distribution at Korle-Bu Teaching Hospital, Ghana. AB - OBJECTIVES: To determine the birthweight distribution of singleton births at the Korle-Bu Teaching Hospital and to determine if selected socio-demographic and reproductive characteristics that are known to be associated with birthweight would show the association in our setting. DESIGN: A non-randomised cross sectional survey of all deliveries within the study period. SETTING: Korle-Bu Teaching Hospital, a tertiary institution, delivering about 11,000 women a year. STUDY POPULATION: From 1st November to 12th December 1994, 866 singleton normally formed livebirths and fresh stillbirths were sequentially enrolled. DATA SOURCES: Data sources were the antenatal and delivery records of the subjects and an interviewer-administered questionnaire. RESULTS: The mean birthweight for the total sample was 3070 g +/- 616 g. One hundred and fifteen (13.3%) babies were low birthweight. The mean birthweight for those with reliable dates and born at term was 3262 g +/- 488.8 g. Multiple logistic regression analysis showed lack of antenatal malaria chemoprophylaxis and a history of previous low birthweight to be significantly associated with low birthweight. CONCLUSION: Although the mean birthweight of Korle-Bu babies was lower than those of USA and UK babies, it was comparable with those from other developing countries. Antenatal malaria chemoprophylaxis is a practical intervention that can produce an increase in mean birthweight and reduce the risk of low birthweight in our population. PMID- 11921566 TI - Fractures of the lateral condyle of the humerus in children. AB - BACKGROUND: Conservative treatment of fractures of the lateral humeral condyle in children is associated with many problems as noted previously by several authors. OBJECTIVE: To determine the outcome of internal fixation of fractures of the lateral humeral condyle. DESIGN: A prospective study. SETTING: Department of Orthopaedic Surgery, Asir Central Hospital, Abha, Saudi Arabia, between 1991 and 1998. SUBJECTS: Sixteen children with fracture of the lateral humeral condyle. All were admitted via the Emergency Room and comprised 14 boys and two girls. Ages ranged between two and 11 years. INTERVENTIONS: All the patients had some form of internal fixation. Percutaneous Kirschner K--wire fixation was done for the undisplaced and minimally displaced fractures and open reduction and K-wire fixation for the widely displaced and rotated fractures. One patient developed non-union of the lateral condyle fracture that required bone grafting and internal fixation with lag screw. MAIN OUTCOME MEASURES: Enhancement of union and reduction of complications resulting from conservative treatment of the fractures. RESULTS: Out of the sixteen cases, there was one case with a minor degree of mal-union on account of loose K-wire and one case with non-union following open reduction and internal fixation. CONCLUSION: The authors recommend internal fixation of all fractures of the lateral humeral condyle in children irrespective of the degree of displacement. PMID- 11921567 TI - Epidemiology of poliomyelitis in northwestern Ethiopia. AB - OBJECTIVES: To determine the magnitude of poliomyelitis and assess its epidemiological features. DESIGN: A cross-sectional community based, house-to house survey. SETTING: Six urban and ten rural villages in Gondar Zuria district, north-western Ethiopia. SUBJECTS: Twelve thousand children aged 1-15 years residing in the randomly selected areas were enrolled in the study to identify children with walking abnormality. Paralytic poliomyelitis is considered as flaccid paralysis in one or both legs with normal sensations and acute onset without progression. RESULTS: Out of 12,000 children aged 1-15 years enrolled, 20 were found to have paralytic poliomyelitis. The prevalence of residual poliomyelitis was found to be 2.1 per 1000 children aged 1-15 years with the estimated annual incidence rate of 7.7 per 100,000 population. Sixty percent of the cases were from urban areas and 55% of the cases were males. CONCLUSIONS: The magnitude of paralytic poliomyelitis is high in the study area, thus expanding the expanded programme of immunizations with a strong surveillance system is suggested. PMID- 11921568 TI - Prevalence of acute and persistent diarrhoea in north Gondar zone, Ethiopia. AB - OBJECTIVES: A study was conducted to determine the magnitude of dysentery and persistent diarrhoea in children aged under-five. DESIGN: A cross-sectional community-based survey was used to enroll the under-five children. A two-stage random sampling technique was applied to identify the households with under-five children. SETTING: The study was conducted from March to May 1997 in two districts of North Gondar Administrative Zone that is located in northwest Ethiopia. Two urban dwellers associations and twelve peasant associations were covered by the study. SUBJECTS: A total of 1101 under-five children were involved. MAIN OUTCOME MEASURES: Bloody diarrhoea and diarrhoea that persisted for fourteen days or more were the main outcome variables. RESULTS: One hundred and ninety seven (18%) of the under five children had diarrhoeal attacks within two-week recall period. Sixty five (33%) of these had persistent diarrhoea. The prevalence of persistent diarrhoea was 6%. Of those who had diarrhoea, forty four (22.3%) had dysentery. Access to protected water source was significantly lower in children with diarrhoeal diseases than without (chi 2 = 4.31, p < 0.05). Significantly a higher number of children with diarrhoea had recent attacks of diarrhoea than those without diarrhoea (chi 2 = 176.82, p < 0.00001). CONCLUSIONS: A particularly high prevalence of dysentery and persistent diarrhoea was observed. Establishment of a simple surveillance system and taking of control measures to reduce the burden of dysentery is recommended. PMID- 11921569 TI - Erythrocyte membrane protein alteration in diabetics. AB - OBJECTIVES: To study the protein components of the erythrocyte membranes of diabetic Nigerians and to compare the results with the erythrocyte membrane protein components of normal healthy Nigerians. DESIGN: Laboratory based cross sectional study. SETTING: Department of Medicine, University College Hospital (UCH), Ibadan and Biomembrane Research Laboratory, Department of Biochemistry, College of Medicine, University of Ibadan, Ibadan, Nigeria. SUBJECTS AND METHODS: The study was carried out in patients with insulin-dependent diabetes mellitus- IDDM (Type 1 diabetes), non-insulin dependent diabetes mellitus--NIDDM (Type 2 diabetes) and healthy human volunteers (HHm), which served as controls. The subjects were aged 30-65 years. There were 12 subjects in each of the IDDM and NIDDM) and 18 subjects in the HHm group. Blood samples (20 ml per subject) were obtained from each subject and erythrocyte ghost membranes were isolated separately from each sample. Total erythrocyte membrane protein concentration of each sample was determined using bovine serum albumin (BSA) as standard. The protein components of the erythrocyte ghost membranes were determined using sodium dodecyl sulphate polyacrylamide gel electrophoresis. STATISTICAL METHOD: All values given are the mean +/- standard deviation (+/- SD) of the parameters measured. Significance was assessed using Student's t-test and P values of 0.05 or less were taken as statistically significant. RESULTS: The total protein concentration of HHm was 5.5 +/- 0.01 micrograms/ml, total protein concentration of IDDM was 4.5 +/- 0.01 micrograms/ml while NIDDM was 5.1 +/- 0.02 micrograms/ml. The spectrin alpha and beta-chain bands are heavily present in the healthy human erythrocyte membranes and are absent in the insulin dependent diabetic membranes. The ankyrin band, band six and below are more pronounced in IDDM and NIDDM but are relatively absent in the healthy humans. CONCLUSIONS: The results obtained provide evidence of profound quantitative and qualitative alteration of the erythrocyte membrane proteins in diabetic Nigerians. This may likely have serious functional implications of the diabetic patients. PMID- 11921570 TI - Experience with road traffic accident victims at The Nairobi Hospital. AB - OBJECTIVE: To evaluate the clinical and epidemiological data on automobile injuries and to assess the adequacy of road trauma documentation at the Nairobi Hospital. DESIGN: A retrospective descriptive study. SETTING: The Accident and Emergency Centre of the Nairobi Hospital. SUBJECTS AND METHOD: Medical records of randomly selected road trauma patients who presented at The Accident Centre between 1st July 1997 and 31st August 1998 were analysed. RESULTS: The mean age was 32 years with a peak incidence in the 21-30 year age group. Males comprised 63.1% of the injured. The predominant category of the road user injured was the vehicle occupant (70%). Pedestrians only constituted 21.3%. Major city roads or highways were the commonest scenes of injury (38.3%). Most of the responsible vehicles were small personal cars (65.8%). The public service minibuses (popularly known as matatu) caused 20% of the injuries. Most of the injuries were mild and transport of the injured to hospital was uniformly haphazard. A quarter of the injuries were severe enough to warrant admission. Trauma documentation was poor with less than 30% accuracy in most parameters. CONCLUSION: The pre-hospital and initial care of the injured is not systematized. The study calls for re orientation of trauma care departments to the care of the injured. PMID- 11921571 TI - Vertebral bone collapse in sickle cell disease: a report of two cases. AB - We describe two female children both nine years of age with sickle cell anaemia and compression deformity of three successive lumbar vertebrae in one child and collapse of one lumbar vertebra in the other. Management for the two children included analgesics, antibiotics and application of a lumbar jacket for stabilisation of the spine. In both patients the vertebral bodies remodelling with re-generation of the tissue. This report is being made to highlight the improved chances of response with early detection and adequate management. PMID- 11921572 TI - Tropical idiopathic lower limb gangrene: case report. AB - Tropical idiopathic lower limb gangrene is a rare disease. It was first described by Gelfand amongst the indigenous inhabitants of present day Zimbabwe. It is a bilateral and simultaneous gangrene of both lower extremities due to no obvious cause and usually seen in men during the second and fourth decade of life. The onset is always sudden and the first sign is oedema of both feet accompanied by pain. The patients are usually people who have been previously healthy. This is a report of a clinical variant of the disease. PMID- 11921573 TI - Micronutrients in health and disease. PMID- 11921574 TI - Vitamin A levels in HIV/AIDS. AB - OBJECTIVE: To study the correlation of vitamin A concentrations in patients with AIDS, HIV positive symptom free and HIV negative symptom free men and women. DESIGN: A cross-sectional study. SUBJECTS: Male and female volunteers aged between 15 and 60 years willing to undergo an HIV-test. SETTING: Participants came from different backgrounds within the city of Ndola. Some were urban while others were peri-urban dwellers. They were included in the study only if they were willing to undergo the HIV test regardless of their place of residence. MAIN OUTCOME MEASURES: After obtaining consent blood samples were taken from the participants using needle and syringe. Whole blood was used to measure haematological indices while serum was used to measure vitamin A concentrations and HIV status. RESULTS: One hundred and thirty five participants were recruited for the study. Vitamin A was analysed in eighty seven HIV negative symptom free, forty one HIV-positive symptom free and seven AIDS cases. There was a significant difference (p < 0.05) in the variance of vitamin A levels in the three groups. Vitamin A deficiency is defined as blood concentrations below 30 micrograms/dl. Using this cut-off point, the Odds Ratio for deficiency if HIV-positive was found to be 6.3 (2.5, 16.7 p < 0.0001). The Odds Ratio for HIV and serum vitamin A deficiency was approximately the same for males and females. There was a modest correlation between vitamin A concentrations and haemoglobin (r = 0.34, 95% CI 0.18, 0.48, p < 0.0001). CONCLUSION: Vitamin A concentration is lowered in HIV infection. The depletion of vitamin A seems to increase with progression of the infection leading to AIDS disease. Whether regular supplementation of vitamin A to the HIV infected individual can lead to a delayed progression to AIDS needs to be explored. PMID- 11921575 TI - Iron and vitamin A status of breastfeeding mothers in Zambia. AB - OBJECTIVE: To determine the prevalence of vitamin A deficiency and iron deficiency anaemia in breastfeeding mothers. DESIGN: Cross sectional descriptive survey. SUBJECTS: Breastfeeding mothers bringing their children to under five clinic. SETTING: A shanty compound in the outskirts of Ndola city, Zambia, Central Africa. MAIN OUTCOME MEASURES: Concentrations of vitamin A, iron, haemoglobin and correlation between vitamin A and iron. RESULTS: Anaemia was found in 29% while iron deficiency was present in 41% of the women studied. Iron deficiency anaemia was responsible for 22% of the anaemia cases. A positive correlation was found between serum vitamin A and haemoglobin concentrations (r = 0.184) and a significant statistical relationship existed (p < 0.05). This statistical significance was even greater when a correlation was made between haemoglobin concentrations and vitamin A concentrations below or equal to 30 micrograms/dl (1.05 mumol/1) (r = +0.351, p < 0.01). CONCLUSION: Vitamin A deficiency and iron deficiency anaemia need to be urgently addressed in this shanty compound and others like it surrounding Ndola city. Supplementing breastfeeding women with vitamin A will not only reduce vitamin A deficiency, but could also help to reduce anaemia. PMID- 11921576 TI - Pathogenic isolates in meningitis patients in Dar Es Salaam, Tanzania. AB - OBJECTIVES: To determine, from laboratory records, the spectrum of bacterial and fungal pathogens isolated from cerebrospinal fluids (CSF) of in-patients with meningitis at Muhimbili Medical Centre (MMC) in Dar es Salaam and to ascertain the laboratory results (based on microscopy and culture) using the latex agglutination technique. DESIGN: A retrospective study based on laboratory records of CSF samples investigated between November 1999 and June 2000 and a cross-sectional study involving investigation of 60 freshly collected CSF samples by conventional (microscopy and culture) and antigen detection by latex agglutination technique (LAT). SETTING: Muhimbili Medical Centre in Dar es Salaam, Tanzania. INVESTIGATIONS: Information from laboratory records, bacteriological examination of CSF by microscopy, culture and agglutination techniques. RESULTS: According to records, a total of 1144 CSF samples were investigated between November 1999 and June 2000, of which two hundred and twenty two specimens (19.4%) had a positive bacterial or fungal culture. Fifty five of the isolates were from children (aged less than 15 years) and were; 20 (36.4%) were Streptococcus pneumoniae, 12(21.8%) were Cryptococcus neoformans, and nine (16.4%) were Haemophilus influenzae type b. The remaining 14 (25%) isolates included three group B streptococci, three Klebsiella spp, two E. coli, two Staphylococcus aureus, two Pseudomonas spp, one Moraxella and one Salmonella group B. For adults a total of 167 positive cultures were reported and 163 (97.6%) of the isolates were Cryptococcus neoformans, two (1.2%) were Pseudomonas spp. and two were S. aureus. There was good agreement between conventional microscopy and culture with the latex agglutination technique in the identification of CSF pathogens. CONCLUSION: In children, S. pneumonia, and bacteria in general constituted the majority of isolates. Adult cases of meningitis were almost exclusively due to C. neoformans. Overall, C. neoformans appears to be the most common isolate among meningitis cases. Based on LAT results, our routine diagnostic methods seem to be adequate in the identification of the common CSF pathogens. PMID- 11921577 TI - Pulmonary functions in ARC-welders in Ile-Ife, Nigeria. AB - OBJECTIVE: To investigate the occupational effects of Arc-welding on small scale welders in Modakeke, Ile-Ife, Nigeria. DESIGN: A community-based, cross-sectional study. SETTING: A semi-urban area Modakeke, Ile-Ife, Nigeria. POPULATION: A semi urban area with a population of about 300,000 (three hundred thousand). STUDY POPULATION: Forty four Arc-welders in Modakeke, Ile-Ife and fifty age and sex matched controls working in the Maintenance Department of Obafemi Awolowo University, Ile-Ife, Nigeria. INTERVENTION: Structured questionnaires adapted from the Medical Research Council, UK respiratory questionnaire with some modifications were administered to all subjects and controls. Physical examination and lung function tests were also carried out on all subjects and controls. MAIN OUTCOME MEASURES: Occupational-related symptoms of welders and controls were compared. Ventilatory function of welders and controls were also compared using the Student's 't'-test. Chi-square was adopted to test the strength of relationship between smoking and lung function abnormalities. RESULTS: The most frequent symptoms found among the subjects were eye irritation (95.43%), rhinitis (50.09%) and skin irritation (43.19%). The arc-welders were found to have characteristically lower lung function parameters than controls (p < 0.05). Ten (22.7%) of the subjects had obstructive lung disease compared to four (8%) of the controls; while 18 (40.9%) of the subjects had restrictive lung disease. Their lung abnormalities had no relationship to their smoking history. CONCLUSION: Workers exposed to arc-welding are at risk of occupational-related symptoms and chronic lung disease. Adoption of protective devices and monitoring of welders' breathing zones for toxic levels of gases and fumes are necessary to reduce the risk associated with arc-welding. PMID- 11921578 TI - Intraoperative blood salvage and autotransfusion in the management of ruptured ectopic pregnancy: a review. AB - OBJECTIVE: To review the role of intraoperative blood salvage and autologous blood transfusion in the management of ruptured ectopic pregnancy. DATA SOURCES: A complete review of relevant current and old literature using the MEDLINE search strategy. STUDY SELECTION: Papers were selected for their relevance to the topic. Preference was for use of blood salvage and autotransfusion in ectopic pregnancy. Other related studies dealing with the use of intraperitoneal blood were also reviewed. DATA EXTRACTION AND SYNTHESIS: Papers were read and analysed. The information on the properties of intraperitoneal blood, the indications, contraindications, complications as well as the techniques for its use were synthesised for the current article. CONCLUSION: Based on reviewed information, intraoperative blood salvage and autotransfusion is a simple, effective and safe method of blood replacement. Its use should be of primary consideration in the management of ruptured ectopic pregnancy. PMID- 11921579 TI - Maternal mortality in Kenya: the state of health facilities in a rural district. AB - BACKGROUND: This study was formulated from the premise that the known causes of maternal mortality, namely haemorrhage, sepsis, obstructed labour and abortion belie the more fundamental development problems that influence it, such as the state of local medical services, quality of care and the facilities' ability to respond to reproductive health emergencies. OBJECTIVE: To document some of the underlying problems and how they were found to influence maternal mortality in Kenya, with specific reference to a rural district. DESIGN: The researchers used the Prevention of Maternal Mortality Network (PMMN) methodology/study design to assess the current state of health facilities, their level of function, and factors influencing their utilisation. Both qualitative and quantitative methods of data collection tools were used. SETTING: Siaya District in the western region of Kenya. Data were collected from thirty facilities, which provide obstetric care in the district. PARTICIPANTS: Data were collected by nurse/midwives, nursing school tutors and social scientists with experience in qualitative research methods. Respondents included health service providers and managers at the 30 health facilities. Qualitative data were obtained through focus group discussions with health facility staff as well as community members. RESULTS: All the thirty facilities studied, were grossly wanting in terms of staffing, equipment, essential drugs and supplies. Both quality of care and record keeping were well below acceptable standards. CONCLUSIONS: The study findings are a sad but a fair reflection of our situation not only in Kenya but also in sub-Saharan Africa ten years after the declaration of the Safe Motherhood Initiative (SMI). The results indicate a predictable, widening gap in basic service provision that must be urgently bridged as a prerequisite to any serious and meaningful approaches to reducing maternal mortality in Africa. PMID- 11921580 TI - Incidence and characteristics of Myasthenia gravis in Dar Es Salaam, Tanzania. AB - OBJECTIVE: To ascertain the annual incidence and characterise the clinical features of Myasthenia gravis in Dar es Salaam, Tanzania during a ten-year period from 1st January 1988 to 31 December 1998. DESIGN: Prospective cumulative registration at a major urban hospital of all patients with newly diagnosed Myasthenia gravis who were resident in Dar es salaam. SETTING: Muhimbili Medical Centre Teaching Hospital, Dar es Salaam, Tanzania. SUBJECTS: Forty seven patients, twenty males, twenty seven females satisfied the criteria for the diagnosis of Myasthenia gravis (MG). RESULTS: The annual incidence of MG of both sexes was 3 per 1,000,000 population of all age groups (95% CI 2.0 to 3.6). The incidence per 1,000,000 population was lowest in those aged below ten years 2.2 (95% CI 1.4 to 3.4) which was statistically significant. The incidence per 1,000,000 per year was higher in females but this was not statistically significant. The clinical presentation of MG in Tanzania was localised disease accounting for 47% ocular type and 53% was mild to moderately severe generalised type MG. Twenty per cent of patients with generalised MG presented with bulbar features. Single fibre electromyography was the most sensitive test. CONCLUSION: Myasthenia gravis is fairly rare in Tanzania as is in other sub-Saharan countries and presents in similar way to European in terms of age, sex, and clinical features. Ocular Myasthenia gravis was more prevalent in Tanzania than in Europe. PMID- 11921581 TI - Bowel resection in children. AB - BACKGROUND: In developed countries, most bowel resections in children are for congenital anomalies and massive resection and short bowel syndrome are frequent problems. OBJECTIVE: To review indications, morbidity and mortality of bowel resection in children in a developing country. DESIGN: A retrospective study. SETTING: Ahmadu Bello University Teaching Hospital Zaria, Nigeria. SUBJECTS: Forty six children aged < or = 12 years who had bowel resection in a period of five years. RESULTS: The commonest indication was typhoid perforation 25 (54%), 21 (84%) of them aged > 5 years. Intussusception was the indication in eight (17%), six less than one year. Bowel resection was performed for congenital anomalies in nine (18%) patients, seven of them < 1 month (abdominal wall defect in four, intestinal atresia in two, strangulating inguinal hernia in one, strangulating band in one, Meckel's diverticulitis, one). Resection for trauma and neoplastic conditions were few (2.4% each). Twelve patients (26%) developed 17 complications including would infection in seven (15%) anastomotic dehiscence, five (11%), abdominal wound dehiscence, three (14%), intra-abdominal abscess and stitch sinus, one (2%) each respectively. Most of the complications followed right hemicolectomy for intussusception and resection for typhoid perforation. Massive bowel resection was necessary in only one patient. There were nine deaths (20%). The mortality was highest in neonates and infants (38%). CONCLUSION: The indications for bowel resection in this environment differ from that in developed countries. Most of the indications (74%) are preventable by improvements in sanitation and early presentation and treatment. The morbidity and mortality are highest in neonates and infants and meticulous attention to technical details may minimise them. PMID- 11921582 TI - Hepatitis B surface antigen and antibody to hepatitis C virus among accident and emergency patients. AB - OBJECTIVE: To determine the sero-prevalence and epidemiology of antibody to hepatitis C virus (anti-HCV) and hepatitis B surface antigen in accident and emergency patients. DESIGN: A descriptive study was performed on 150 accident and emergency patients. Sera was screened for anti-HCV and HbsAg, using enzyme linked immunosorbent assay. Personal information and risk factors involved were obtained using a questionnaire. SETTING: Haematology laboratory of the University of Benin Teaching Hospital, Nigeria. SUBJECTS: One hundred and fifty adults consisting of 122 males and 28 females who were above the age of 21 years. Patients were randomly selected from all adults including dental patients attending the emergency department for both surgical, dental and medical emergencies. The age range was between 21-58 years. In order to ascertain the epidemiology of both viruses a questionnaire was used detailing the possible risk factors for transmission. These included history of previous blood transfusion; history of life time occurrence of multiple sexually transmitted diseases; history of heterosexual exposure to partners at risk (for example prostitutes); history involving the use of unsterilized blades; presence of scarification marks and tattooing; low socio-economic status (low annual income or unemployed): history of intravenous drug use and heterosexual activity. Anti-HCV and HBsAg were both assayed using different assay kits, based on the enzyme linked immuno absorbent assay (ELISA) tests from different manufacturers. RESULTS: The sero-prevalence of anti-HCV was 14% and 29.3% for HBsAg. Anti-HCV positivity was significantly associated with a past history of blood transfusion and heterosexual exposure to partners at risk. The study also revealed a significant association between HBsAg positivity and all the risk factors. CONCLUSION: The high prevalence rate for anti-HCV and HbsAg in accident and emergency patients increases the likelihood of further spread from patients to health care providers especially if adequate precautions are not observed. PMID- 11921583 TI - Incidental mucinous cystadenocarcinoma of the appendix with pseudomyxoma peritoni in a diabetic with hypoglycaemic coma. PMID- 11921584 TI - Serum leptin concentration in a rural African population. AB - OBJECTIVE: To determine the serum leptin concentration in a cohort of healthy rural Africans, it's relation to the commonly used anthropometric measures of obesity and its relation to the patterns of distribution of fat in the body. DESIGN: A cross-sectional population survey. SETTING: Baziya area, Transkei region, South Africa. SUBJECTS: One hundred and thirty five (79 females and 56 males) healthy adults from the Baziya location, Transkei aged 17-70 years were selected by stratified random sampling. MEASUREMENTS: Anthropometric measurements of height, weight, and skinfold thickness at the biceps, triceps, subscapular and suprailiac sites and derived total body fat and ratios of fat distribution. Fasting serum leptin using the sandwich ELISA method. RESULTS: Skinfold measurement was significantly higher in the females than the males throughout the age range. Centralization of body fat to the trunk was significantly greater in the males than in the females. Serum leptin concentration was higher in the females (mean = 13.5 ng/ml; 95% confidence interval = 10.0-16.8) than in the males (mean = 5.2 ng/ml; 95% confidence interval = 2.8-7.6) (p < 0.001). The gender difference in leptin concentration persists when expressed as serum leptin per kilogram of fat mass (serum leptin (ng/ml)/FM). The mean value for the males was 5.1 ng/ml/kg (95% confidence interval = 2.9-7.3) compared to the mean value for females of 6.9 ng/ml/kg (95% confidence interval = 5.4-8.3) (p < 0.05). In the females BMI and body fat were significant contributors to the variance in serum leptin. In the males the upper-to-lower trunk skinfold thickness ratio and BMI were the significant contributors to the variance in serum leptin concentration. Deposition of fat in the abdomen did not have a significant contribution to the variance in circulating leptin in both sexes. CONCLUSION: Serum leptin concentration in rural Africans is similar to that observed in other communities with the exception that regional fat distribution has a significant influence on the leptin levels in the males. PMID- 11921586 TI - Suicide attempts in a Nigerian military setting. AB - OBJECTIVES: To describe the characteristics of patients who attempted suicide in a military setting and to examine the differences between the suicide attempters and a group of non-suicide, affective disorder patients. DESIGN: Fifty one consecutive cases of suicide attempts were studied using a questionnaire which inquired about demographic characteristics and suicide related issues such as method of attempt used and giving a notice. The suicide attempt group was then compared with a control group who had no history of attempted suicide. SETTING: Department of Psychiatry, Military Hospital, Yaba, Lagos, Nigeria, which is a 500 bed military general hospital. SUBJECTS: Fifty one attempted suicide patients. Intervention reports on intervention methods applied were not within the scope of this report. MAIN OUTCOME MEASURES: These were also not within the scope of the present study. RESULTS: Suicide attempt patients constituted 0.37% of all admissions during a five-year period, and 60.8% of them were under the age of 30 years. The numbers of male and female patients were approximately the same. Depression and acute stress reaction were the commonest diagnoses. While military dependents most commonly ingested substances in their suicide attempts, military personnel most often used more violent methods including hanging and self stabbing, but none used firearms. Compared with non suicide patients, suicide attempt ones were more likely to be unmarried and to have a family history of mental disorder but less likely to have lost a parent through death before the age of 18 years. CONCLUSIONS: The need for a more controlled access to substances capable of being used for self-destruction was highlighted. The need for emergency room doctors to become more skilfull in the identification and assessment of suicide attempt patients was also emphasized. PMID- 11921585 TI - Health status of hospitalised elderly in Morogoro Regional Hospital. AB - OBJECTIVE: To assess the elderly's nutritional status, major health and socio economic problems. DESIGN: A cross-sectional, descriptive study. SETTING: Hospitalised care, in Morogoro General Hospital, Morogoro, Tanzania. SUBJECTS: One hundred and twenty one elderly patients admitted in the hospital for various health problems between November 1998 and March 1999. RESULTS: Body Mass Index (BMI) ratios indicated that 26% of the subjects were suffering from Chronic Energy Deficiency (CED) of varying grades. Fourteen percent had CED grade I, 10% CED grade II and two per cent had CED grade III. The study ranked cardiovascular disorders and respiratory tract diseases the most prevalent diseases (30% and 28% respectively). These were followed by malaria (15%). Cardiovascular diseases were more prevalent among women than men while the respiratory diseases were opposite. The general tradition that African elderly people do live with and are taken care by their children seems to hold no longer, as the majority of the subjects (80%) either live alone or with their spouses only, while 14% lived with their relatives (including their children). The majority of the elderly did not have any regular income, and they depended on other people for their living. CONCLUSION: As Tanzania undergoes demographic and economic transformations, the elderly are increasing while their welfare decreases. This has more effects on their health status. Old tradition of children living with their elderly is declining due to urbanisation, rural urban migration and the trend towards nuclear families. The government is urged to establish specific systems of taking care of this vulnerable group. PMID- 11921587 TI - Childhood achalasia in Zaria, Nigeria. AB - OBJECTIVES: To study the presentation, management and outcome of achalasia of the oesophagus in children. DESIGN: A retrospective case study. SETTING: Ahmadu Bello University Teaching Hospital, Zaria, Nigeria. SUBJECTS: Seven children managed for achalasia of the oesophagus in a period of 19 years. RESULTS: There were six boys and one girl. The median age at presentation was ten years (range three months to fifteen years). The median duration of symptoms before presentation was 20 months (range: 2-24 months). Three children were treated on several occasions for pneumonia before the diagnosis was made. Dysphagia, vomiting and regurgitation were the main presenting symptoms. Diagnosis was by barium oesophagogram. Six had modified Heller's myotomy via celiotomy and a Thal fundoplication in a three month old. Transthoracic Heller's myotomy was done in the seventh child. There was complete resolution of symptoms in six children. One patient had post-operative retrosternal pain, which was controlled by oral nifedipine. CONCLUSION: Achalasia of the oesophagus should be excluded in children with recurrent chest infection. Modified Heller's myotomy without antireflux procedure via the abdominal route is effective in relieving symptoms of achalasia in children. PMID- 11921588 TI - Undiagnosed renal abscess presenting as acute bacterial peritonitis: case report. AB - A case of undiagnosed renal abscess complicated by intraperitoneal rupture with generalised bacterial peritonitis is presented. Prompt surgical intervention was essential for saving the patient's life. Early imaging of the kidney in unresolved renal infection is essential if renal abscess, its delayed presentation and complications are to be avoided. Most patients will be cured without operation by antibiotics and if necessary, by additional percutaneous drainage where an abscess has formed. PMID- 11921590 TI - Clinical drug interactions. PMID- 11921589 TI - Re: Prevalence of mycoplasma pnuemoniae in children with acute lower respiratory tract infection. PMID- 11921591 TI - Effects of non-steroidal anti-inflammatory drugs on hypertension control using angiotensin converting enzyme inhibitors and thiazide diuretics. AB - OBJECTIVES: To determine the impact of three non-steroidal anti-inflammatory drugs on the efficacy of two anti-hypertensive drugs. DESIGN: Fifteen women with arthritis and hypertension who were receiving lisinopril and HCT, and administered sequentially in random order ibuprofen, sulindac, and diclofenac for one month each, with an intervening two-week washout period between each treatment period. During the washout period, subjects received paracetamol. SETTING: Hypertension Clinic, Medical Centre, Harare, Zimbabwe. SUBJECTS: Fifteen female hypertensive women with documented arthritis. MAIN OUTCOME MEASURES: Blood pressure at the end of two weeks of paracetamol was compared with blood pressure after one month of treatment with each of the NSAID. RESULTS: Mean blood pressure was unchanged before and after all NSAIDs: 108 +/- 7 versus 107 +/- 9 for diclofenac, 108 +/- 9 versus 108 +/- 9 for sulindac, and 108 +/- 8 versus 107 +/- 9 for ibuprofen. The 24 hour urinary sodium excretion was not significantly different. CONCLUSION: The three NSAIDs investigated did not neutralise the antihypertensive effect of the combination of lisinopril and HCT, and hence the blood pressure lowering action of the combination may not be prostaglandin dependent. PMID- 11921593 TI - Validation of reports of nodules dissolution after repeated ivermectin treatment of onchocerciasis in southeastern Nigeria. AB - OBJECTIVE: To ascertain the validity of the claims by villagers in endemic communities in southeastern Nigeria that repeated ivermectin treatment cause disappearance or dissolution of onchocercal nodules. DESIGN: From baseline epidemiological data, four communities (Amuro, Aku, Nzerem Ikpem and Awuchinimo) were randomly chosen as the study sites. A total of 324 adults (hitherto had onchocercal nodules), aged > or = 20 years from these communities were enlisted in the study. After clinical examination, each of the villager was interviewed with structured, pre-tested questionnaire. Also personal and focus group discussion was held. RESULTS: Sixty nine (21.30%) respondents; 16.67% from Amuro, 32.26% from Aku, 19.05% from Nzerem Ikpem and 15% from Awuchnimo claimed that their nodules gradually dissolved or disappeared. A significant (p < 0.05) proportion (82.61%) attributed this phenomenon to repeated doses of ivermectin; 4.35% claimed it was not due to ivermectin therapy while 13.04% were not sure of the cause. CONCLUSION: The results have opened challenges in our understanding of the role of ivermectin therapy in endemic communities and calls for further studies of nodule dissolution in other onchocerciasis endemic communities. PMID- 11921592 TI - Prescribing habits in church-owned primary health care facilities in Dar Es Salaam and other Tanzanian coast regions. AB - OBJECTIVE: To assess prescribing practice of Primary Health Care (PHC) workers in church owned health care facilities using WHO drug use indicators. DESIGN: A cross-sectional study in which twenty primary health care facilities were randomly selected. Prescribing indicators were obtained by analysing outpatient records retrospectively for the past 14 months between January 1997 and February 1998. This period was chosen because of compete records of outpatient attendances. Patient care and facility indicators were recorded prospectively during the study period. SETTING: The study was conducted in the Coast and Dar es Salaam regions of Tanzania. Six districts were randomly selected from both regions. The selected districts included Ilala, Temeke and Kinondoni in Dar es Salaam, Kibiti, Bagamoyo and Kisarawe in Coast region. SUBJECTS/MATERIALS: Twenty primary health care facilities were randomly selected from the chosen districts. Patient registers were collected and patients' characteristics including age, sex, diagnosis, and drugs prescribed for the period January 1997 to February 1998 were recorded on data collection forms. Patient care indicators were measured by recording consultation time, dispensing time, per cent of drugs actually dispensed and adequately labelled whereas patients' knowledge of correct drug dosage was obtained using exit interviews. Verification of facility indicators was done by direct observation. RESULTS: The average number of drugs per prescription was 2.3 (range 1.8-2.8). Generic prescribing prevailed with a mean of 75.5% of all drugs. Antibiotic and injection encounters per prescription was 35.4 and 19%, respectively. Most drugs were prescribed according to the essential drug list of Tanzania (NEDLIT). Patient's average consultation time was 3.6 minutes whereas average dispensing time was 39.9 seconds. On average, 87% of all drugs dispensed were adequately labelled and patients' knowledge of correct dose was adequate. All facilities possessed drugs for treating important illnesses, all had reference educational materials. CONCLUSION: The study shows that there is an overuse of injections 19% +/- 1.7 (range 0.73%) compared to the recommended figure of 15%. The use of antibiotics appears appropriate when compared with the morbidity patterns in the study areas. A focus group discussion with prescribers in these facilities to address the question of overuse of injections is needed in order to plan an appropriate intervention. PMID- 11921594 TI - Antimicrobial susceptibility and plasmids from Escherichia coli isolated from rats. AB - OBJECTIVES: To determine if antimicrobial resistance occurs in E. coli isolated from rats and if this resistance is transferable via plasmids. METHODS: Sixty Escherichia coli isolates obtained from 215 rats trapped in and around Nairobi, Kenya, were analysed for antimicrobial susceptibility to eleven commonly used antimicrobials. Plasmid DNA analysis and in-vitro conjugation tests were conducted on twenty two resistant isolates. RESULTS: A total of 12 (20%) E. coli were resistant to at least one of the commonly used drugs including ampicillin, sulphamethoxazole and streptomycin. Eight (13.3%) isolates were multidrug resistant. The drug resistant E. coli fell into five plasmid profile groups with plasmids ranging between two and 98 megadaltons (Mda). Resistance to ampicillin was transferable. CONCLUSION: The results of this study show that rats from the study area may form an important reservoir for drug-resistant E. coli that could pose a public health hazard. PMID- 11921595 TI - Molecular epidemiology of Bacillus cereus food poisoning. AB - OBJECTIVES: To investigate the potential use of DNA techniques in epidemiological diagnosis of Bacillus cereus food poisoning. SUBJECTS: Fifty six B. cereus isolates from milk were studied. DESIGN: The 56 B. cereus isolates were characterised into enterotoxin positive (27 isolates) and enterotoxin negative (29 isolates) using reverse passive latex agglutination technique. SETTING: Plasmid and genomic DNA were isolated from all the B. cereus isolates. The plasmid DNA was analysed by gel electrophoresis, while genomic DNA was used for restriction endonuclease and toxin gene analyses. MAIN OUTCOME MEASURES: Plasmid profile analysis, restriction endonuclease analysis of genomic DNA, and test for bceT and hblA genes by polymerase chain reaction and gene probing. RESULTS: Seventy two per cent of the isolates contained one to five plasmids of molecular sizes between 0.1 to 60 mDa. Restriction analysis of genomic DNA gave different restriction patterns among enterotoxin positive and enterotoxin negative isolates. A polymerase chain reaction assay detected bceT gene in 41.1% of the isolates, 16% of which tested positive for enterotoxin with B. cereus enterotoxin reverse passive latex agglutination (BCET-RPLA) kit, while hblA gene was detected in all the enterotoxin positive isolates. BceT and hblA gene probes detected the respective genes in all the isolates that also tested positive for toxin genes by polymerase chain reaction. CONCLUSION: DNA techniques provide an alternative approach to the diagnosis of enterotoxigenic B. cereus. PMID- 11921596 TI - Thirty day mortality and related variables in open heart patients at the Kenyatta National Hospital, Nairobi. AB - OBJECTIVE: To determine the thirty-day mortality of open-heart patients at the Kenyatta National Hospital in Nairobi from June 1973 to October 2000 and; to look at likely variables related to mortality. DESIGN: A retrospective analysis of data from the unit database. Data on this database were collected prospectively from September 1997 to the time of study. Data in respect of the period prior to this were collected retrospectively from patient files, ward and theatre records. SETTING: Kenyatta National Hospital (KNH), Nairobi. PATIENTS: A total of 563 open heart patients operated at the KNH were included in the study. RESULTS: The thirty-day mortality rate calculated at 17.4% for the study period compared to a hospital mortality rate of 16.9%. Surgical repair for complex congenital pathology, surgery on patients with a left atrial (LA) dimension or a left ventricular end systolic dimension (LVESD) greater than 5 cm or/and a cross clamp time greater than 60 minutes all had a significantly greater risk of mortality on bivariate analysis. This is compared to surgery for simple hole in the heart, LA and LVESD dimensions less than 5 cm and cross clamp times less than 60 minutes (p < 0.05). The increased risk of mortality with these variables was 3.33, 3.95, 3.18 and 1.8 times greater than their counterparts, respectively. For patients having surgery for an acquired pathology, only a cross clamp time greater than 60 minutes and a left atrial size greater than 5 cm were independent risk factors for thirty day mortality using logistic regression analysis. For patients having surgery for correction of a congenital defect, only a cross clamp time of more than 60 minutes was an independent predictor of mortality (p < 0.05). CONCLUSIONS: The higher mortality rate is amongst others, probably related to the late presentation of our patients for surgery when their myocardial function is below the optimum for surgery. There is a need to bring down the mortality through more stringent patient selection, preoperative preparation and reduction of surgical ischaemic times, however without depriving the patients in need of surgery. PMID- 11921597 TI - Aetiology of tinea capitis in school children. AB - BACKGROUND: Tinea capitis is a superficial fungal infection of the scalp and hair of the head which is common in children aged between two and eleven years. An endothrix infection is best treated using parenteral antifungal drugs while an ectothrix one may be treated by parenteral or topical antifungal drugs. The organism that commonly causes tinea capitis in the Western world is Trichophyton tonsurans while there is inadequate information on the actual causative agent in Kenya. OBJECTIVES: To determine the prevalence and the aetiology of tinea capitis and the fungal agents responsible for it and; to recommend the correct mode of treatment in school children attending a primary school near Eldoret. DESIGN: Prospective and descriptive study. SETTING: Private primary school near Eldoret town. SUBJECTS: Sixty eight primary school children aged between six and 14 years in classes 1 to 5. RESULTS: Of the 68 pupils included in the study, 60.9% and 39.1% were males and females, respectively. Prevalence of tinea capitis in the school was 33.3%. Peak age of infection was 10 years. Ratio of infected males to females was 2:1. There was a growth on culture in 76.1% of the cases all of which were endothrix. T. tonsurans was isolated in 77.8%, T. rubrum in four per cent of the cases. CONCLUSION: The prevalence of tinea capitis in this school was high. The most common cause of tinea capitis was T. tonsurans. Males were more infected than the females. RECOMMENDATIONS: Oral antifungal drugs with griseofulvin being the first line of treatment should be adopted. Health education on the aetiology, treatment and prevention of tinea capitis should be given. PMID- 11921598 TI - Plantar and digital dermatoglyphic characteristics of Zimbabwean subjects. AB - OBJECTIVE: To establish the dermatoglyphic characteristics of indigenous Zimbabwean subjects. DESIGN: Cross-sectional study of randomly selected subjects. SETTING: Mufakose, a high-density township in cosmopolitan Harare and Gweru, a midland city in Zimbabwe. MAIN OUTCOME: Dankmeijer (DI) and pattern intensity (PII) indices and the variability of ridge patterns. MATERIALS AND METHODS: Bilateral plantar and digital prints of the sole of selected subjects were recorded, studied and classified. RESULTS: Plantar pattern types, showed more loops than arches and more arches than whorls. The loops and arches were commoner on the distal than proximal zones of the sole; features also shown in Malawians. Whorls were absent in zone II and this appears to be peculiar to Zimbabweans. Digital pattern types showed the predominance of arches as was the case with Malawians but loops were the most prominent in the first digit and next in overall prominence to arches. The mean PII was higher in males than females while DI was higher in females than males, which were also the case with Malawian subjects previously studied. However, for both sexes the PII was significantly higher in Zimbabweans than Malawians while DI was significantly lower in Zimbabweans than Malawians (p < 0.001). CONCLUSION: This study has elucidated the normal dermatoglyphic characteristics of Zimbabweans, showing features that indicate affinities with Malawians. The digital features, however, could differentiate Zimbabweans better from Malawians, further emphasising the uniqueness of digital ridge patterns in differentiating population groups. PMID- 11921599 TI - Septicaemia in high risk neonates at a teaching hospital in Ile-Ife, Nigeria. AB - OBJECTIVES: To determine the incidence, predisposing factors, clinical features, bacteriological pattern and antibiotic sensitivity in septicaemia in high-risk newborns. DESIGN: A prospective study. SETTING: Neonatal unit, Ife State Hospital, a unit of the Obafemi Awolowo University Teaching Hospital Complex (OAUTHC), Ile-Ife, Nigeria. SUBJECTS: All newborns admitted with clinical features and/or risk factors suggestive of neonatal septicaemia from February 1994 to March 1995. MAIN OUTCOME MEASURES: Culture results and mortality rates. RESULTS: The incidence of neonatal septicaemia among new born was 22.9 per 1000 livebirths. The predisposing perinatal factors were low socio-economic status, lack of antenatal care, maternal peripartum pyrexia and congenital malformations. Gram-positive bacteria were found to be the most prevalent causative organisms (59.4%). Staphylococcus aureus (36.2%), Pseudomonas aeruginosa (18.8%) and Coagulase negative Staphylococcus (15.9%) were the commonest causes of septicaemia. Meningitis and UTI were associated diagnoses in 16.7% and 18.2% of the septicaemic babies, respectively. The bacterial isolates showed a high degree of in-vitro antimicrobial resistance. However, all the isolates were sensitive to ofloxacin. Amongst the commonly used antibiotics, gentamicin had the lowest resistance. The overall mortality rate was 33.3%. CONCLUSION: Improvement in the socio-economic status of the populace and availability of affordable antenatal care would reduce the incidence of neonatal septicaemia in Nigeria. Continuous surveillance in every unit, as well as close attention to preventive strategies would be necessary to reduce morbidity and mortality from neonatal septicaemia. We recommend the inclusion of gentamicin in the initial treatment of septicaemia in the neonatal unit of OAUTHC, Ile-Ife, Nigeria. PMID- 11921600 TI - Perinatal morbidity at the Moi Teaching and Referral Hospital, Eldoret. AB - BACKGROUND: Perinatal morbidity (PNM) is highest in the developing countries including Kenya. Studies on the perinatal morbidity in Moi Teaching and Referral Hospital (MTRH) have not been carried out. Furthermore, factors associated with PNM are unknown at the MTRH. OBJECTIVE: To establish the causes and incidence of perinatal morbidity at the MTRH NewBorn Unit (NBU). DESIGN: A retrospective study using case notes. SETTING: The NBU of the MTRH in Eldoret, Kenya. SUBJECTS: Babies who were admitted to the MTRH's Newborn Unit from January to December 1997. RESULTS: The perinatal morbidity was 667 per 1000 babies admitted to the NBU. The most common reason for admission was mother under anaesthesia. The most common cause of morbidity was asphyxia. The mean maternal age was 24 years. The mean level of education was 1.7 years. More than fifty five per cent of the mothers were housewives. The mean birthweight was 2.75 kg (+/- 0.9 SD), 55.9% had normal birthweight, 37% were low birthweight and three per cent were large babies. The mean duration of stay in the NBU was 3.8 days. CONCLUSION: Most of the admissions were Caesarean section babies who did not require admission. Younger mothers were more likely to have babies with neonatal sepsis, convulsions, pneumonia and asphyxia. PMID- 11921602 TI - Domestic violence and induced-abortion: report of three cases. AB - Domestic violence not only violates women's rights, but is also associated with adverse consequences ranging from physical injury to psychological morbidity. Several studies have associated domestic violence with risk of having an induced abortion. The following three cases indicate that domestic violence may not cause unwanted pregnancy, but may be the reason that leads victims to resort to abortion. PMID- 11921601 TI - Good clinical practice in east Africa: a review. AB - OBJECTIVE: To call for harmonisation of good clinical practice guidelines in East Africa as one of the necessary precedents to enhancing their quality and quantity in the region. DATA SOURCES: There were two main sources of background information for this review. The first was a series of articles discussing and documenting the harmonisation process in East Asia and in Latin America. The other was a short survey conducted in Kenya in July 2000. STUDY AREA SELECTION: Most of the survey participants were drawn from the Kenya Medical Research Institute (KEMRI) in Nairobi, Kisumu and Kilifi with a few coming from the vicinity of these areas. DATA PROCESSING AND ANALYSIS: The survey tool was a fourteen page questionnaire seeking information in various areas. These were: personal information, general clinical research information, medical practitioners, clinical research experience, human subject participation, sponsors, clinical trial supplies, investigator sites and general comments. Data entry was performed in Microsoft Access. Forty four researchers participated in a period of three weeks. Data summaries and reports were performed in SAS. CONCLUSION: The advancing capacity for clinical research in East Africa will be accelerated by harmonising clinical research requirements and guidelines. PMID- 11921603 TI - Non-functioning ovarian fibroma with extensive calcification: case report. AB - A case of a non-functioning ovarian fibroma with extensive calcification in a young Ugandan female is described. Ovarian fibromas occasionally may be bilateral and associated with benign ascites and pleural effusion called Meigs' syndrome or related to a rare hereditary condition known as Gorlin's syndrome. Rarely if functioning, the tumour may produce hormones to cause diabetes mellitus or hypoglycaemia or secrete carbohydrate antigen 125 to clinically simulate ovarian carcinoma. PMID- 11921604 TI - Morgagni hernia: case report. AB - This is a case report of an elderly woman who presented with a history of epigastric pain and persistent vomiting diagnosed initially as a duodenal ulcer, later as a pyloric stenosis and at laparotomy was found to have an anterior diaphragmatic hernia with gastric volvulus. Hernia of Morgagni occurs through a congenital defect in the diaphragm but usually presents in adulthood. It could be an incidental diagnosis or can present with obstructing symptoms of the herniated viscera. Treatment is surgical with reduction of hernia and repair of the diaphragmatic defect. If misdiagnosed, this can lead to considerable morbidity and occasionally mortality due to the obstructed/strangulated hernial contents. PMID- 11921605 TI - Epidemiology of retinitis pigmentosa in Denmark. AB - A nation-wide registration of Danish cases of retinitis pigmentosa (RP) provided 1890 persons diagnosed during the period 1850-1989. Prevalent at 1 January 1988 were 1301 persons (1:3943) comprising a multitude of different RP-types. Age specific prevalence rates demonstrated increasing rates of RP during the first four decades of life and a rather stable prevalence over the next 20-30 years. Corrected for incompleteness, a late decrease was found, reflecting an incomplete ascertainment of the oldest patients. A moving average method indicated an even later steady state value for the age-specific prevalence. The Danish prevalence figures were standardized according to the WHO World Standardized Prevalence Rates and compared with large studies from the USA and UK. No statistically significant difference was found. Usher syndrome was present in 12% of all RP cases and Bardet-Biedl syndrome comprised 5%. Mental retardation was found in 144 cases (11%), mostly characterized by atypical RP. Nineteen per cent of patients affected by nonsystemic RP had an onset later than 30 years of age, whereas only a few per cent of persons with systemic RP had an RP onset after age 30 years. The Mendelian inheritance type of all cases was evaluated according to an unambiguous genetic classification, finding a larger amount of X-linked RP compared with other studies. Among nonsystemic RP-cases, 14.3% were found to be inherited as an X-linked trait whereas only 8.4% were autosomal dominantly inherited. The largest fraction was, as in previous materials, the simplex group (isolated cases) comprising 42.9% of the nonsystemic RP patients. Some factors influencing the results are discussed, with special emphasis on the problems associated with precise definitions of the Mendelian inheritance groups. A diagram according to the author's definition was constructed as a guideline ready for clinical application. PMID- 11921606 TI - [Effectiveness of structural quality in quality assurance--a review]. AB - Minimum standards as a part of structural quality are often discussed for the sake of improving the quality of medical care. Before implementing obligatory standards, however, the effectiveness of the demands made should be evaluated. THE METHOD: Our method was a systematrix review (with an eye to structural quality indicators) of literature found by searching Medline; the structural quality indicators taken into account were special forms of medical care, hospital characteristics, certification, internal quality management, internal peer-reviewing, telemedicine, continuing medical education, the use of guidelines, and the caseloads of physicians and hospitals? THE RESULTS: A minimum caseload, the use of guidelines and continuing medical education show positive effects on the outcome of care. The other items show mixed study results or are not measurable in a sense that would make their results of use for quality improvement. Without evidence of effectiveness, minimum standards should not be introduced. Despite the inhomogeneity of the methods used by the studies, minimum caseloads for some diagnoses, the use of guidelines and well-organised continuing medical education are to be recommended. PMID- 11921607 TI - [Aspects of colchicine therapy. 2. Additional classical indications and new therapeutic aspects]. AB - The complex actions of colchicine, which are attributable to its stabilising action on the cytoskeleton and cell membranes and its special pattern of distribution, form the basis of the results presented here; results that relating to the prophylactic and/or therapeutic actions of colchicine in a whole range of other diseases. PMID- 11921608 TI - [Continuing medical education as an "event"--innovative use of satellite technology during information transfer]. AB - While it is true that satellite technology has for some time allowed experts that are physically far apart to congregate through their telepresences in--as a rule- one auditorium, nevertheless, the expenditure of effort involved is rarely worth the attention generated by the medium. Therefore by using innovative and appropriately timed satellite technology a multimedia concept with both the character of an "event" and the lasting quality of an "experience" was developed. This concept is based among other things on the complex interconnecting of four experts in four separate halls delivering four lectures (with ensuing discussions) as well as technically speaking on a live television outside broadcast standard including professional presentation and direction. The implementation of the concept was tested and optimised in the course of two information transfer events. Besides confirming the exceptional quality and motivational benefit of the concept the participants' evaluation also underscored the importance of the professional production with its use of specific forms of presentation (such as interviews). PMID- 11921609 TI - [www.evidence.de--assessment and utilization of clinical guidelines by primary care physicians and internists]. AB - BACKGROUND AND OBJECTIVE: Compared to the generation of new scientific results, implementation and utilization of evidence based knowledge is lacking far behind. Strengths and weaknesses of evidence- and HTML-based guidelines were evaluated in an academic network of primary care physicians to better understand the expectations and information needs of GPs. METHODS: Standardized survey of 72 primary care physicians (21% female, 79% male) regarding three network based issues: "frequency of guideline use", "reasons to use/not to use guidelines", "evaluation and overall judgement of guidelines". RESULTS: 62.9% of participating had used net based guidelines at least once, the percentage of more frequent users (> 2 times in two months) was 40.3%. Curiosity and non-specific interest as well as a concrete medical question were the main reasons for utilization of guidelines. The overall judgement of the guidelines was positive, however, only a minority of participating physicians viewed them as a concrete help in daily practice. CONCLUSION: Transformation of new scientific medical knowledge into daily primary care practice is only insufficiently developed. Netbased instruments of continuing medical education may serve as catalysts and should be further developed as well as evaluated. PMID- 11921610 TI - [Experiences with and impact of health technology assessment on the German Standing Committee of physicians and patients]. AB - The "Bundesausschuss der Aerzte und Krankenkassen" (hereafter referred to as the "Bundesausschuss") is one of Germany's self-governing bodies in ambulatory health care. This Federal Standing Committee of Physicians and Sickness Funds has the mandate, among others, to issue directives for the ambulatory health care provided by 110,000 physicians to the approximately 70 million citizens of Germany. The Bundesausschuss' directives are legally binding and must be followed by both the physicians who provide ambulatory medical services and the sickness funds who insure these services. Although elements of evidence-based medicine were first introduced in 1990, health technology assessment (HTA) was not systematically integrated into the decision making process of the working group for medical and surgical procedures until 1998. The HTA reports of the Bundesausschuss take account of the status of the technology in other health care systems (public systems in particular) and of statements by the scientific community and interested parties, such as patient groups, and review the evidence currently available (in primary studies, systematic reviews, other assessments, etc.). These reports are prepared by representatives of the physicians and the sickness funds, in close collaboration, whenever possible, with academic experts in health technology assessment. The reports serve as a basis for the decisions made by the Bundesausschuss, which ensures their impact, and are published on the Internet at http://www.kbv.de/hta. The Bundesausschuss reports also reflect the deliberation process for each technology, in the context of the legal environment. They also explain how a decision was reached. The shortcomings of HTA process are the time and financial resources required to conduct these assessment. The advantages include the increased transparency of the decision making process, for both the German "social" court system and for the insured. Further developments will be directed by the future framing of the legal environment by the ministry of health. A similar decision-making body has recently been installed for the hospital sector and both committees will be co ordinated by a board, which will organize the deliberation process. It will be important to further encourage and integrate the use of evidence in the decision making processes of these statutory bodies. PMID- 11921611 TI - [Assessment of methodological quality of clinical performance measures]. AB - One of the tasks of the federal "Coordinating Committee" within Germany's self governing health care system is to develop clinical performance measures. Since these performance measures usually exert an enormous influence on health care delivery their methodological quality should meet highest standards. The aim of our study was to develop quality requirements for clinical performance measures to be used in Germany; we did this mainly by compiling internationally published criteria. We present lists of quality requirements for clinical performance measures and performance assessment programs as well as a checklist for documenting and assessing comprehensively the methodological quality of these measures. Developing clinical performance measures by using these criteria could improve their methodological quality and promote the objectivity of performance assessment in medical care. PMID- 11921612 TI - [Meta-analyses of quarks, baryons and mesons--a "Cochrane Collaboration" in particle physics]. AB - Within the last 20 years meta-analysis has become an important research technique in medicine for integrating the results of independent studies. Meta-analytical techniques, however, are much older. In particle physics for 50 years now the properties of huge numbers of particles have been assessed in meta-analyses. The Cochrane Collaboration's counterpart in physics is the Particle Data Group. This article compares methodological and organisational aspects of meta-analyses in medicine and physics. Several interesting parallels exist, especially with regard to methodology. PMID- 11921613 TI - Recruitment. On the Bengal beat. PMID- 11921614 TI - Sexual health. Fighting a false sense of security. PMID- 11921615 TI - Older people. The age of contempt. PMID- 11921616 TI - Does nursing get the leadership it deserves? PMID- 11921617 TI - General nurses are special too. PMID- 11921619 TI - Total courage. PMID- 11921618 TI - A hard day's night. PMID- 11921620 TI - Epilepsy liaison nursing. AB - The epilepsy liaison nurse's aim is to achieve a positive outcome in terms of clients' general well-being, social and physical activities, behavioural difficulties and other important issues in their lives. Many clients' problems stem from a lack of adequate information and support. A sound knowledge of the disorder can boost clients' and carers' confidence. They need information on what to do during a seizure, how to ensure that other people know what to do, and to understand the importance of being aware of precipitating factors. Clients and their families should also be encouraged to talk about the condition. This will reveal any misconceptions and anxieties and bring concerns into the open, helping to put them into perspective. Support for carers includes ensuring that clients have the personal care and support they need to improve or maintain a good quality of life. It is important that the person managing the client's epilepsy understands any issues related to learning disabilities and knows how to gain access to health and social care systems. By identifying and addressing the needs of clients and their families, epilepsy liaison nurses can make a positive difference to their lives. PMID- 11921621 TI - How faxes improve contact between care settings. PMID- 11921622 TI - Nursing with dignity. Part 2: Buddhism. PMID- 11921623 TI - Support for pregnant women. PMID- 11921624 TI - Workforce planning. Doctors propose radical overhaul of nurses' role. PMID- 11921625 TI - Rheumatology. 3. Modern drug therapies and current guidelines in rheumatology nursing. PMID- 11921626 TI - A managed clinical network for home parenteral nutrition. PMID- 11921627 TI - Therapeutic interventions in osteoporosis. PMID- 11921629 TI - Pressures and poverty drive students to therapy. PMID- 11921630 TI - Scottish patients still wait more than a month for admission. PMID- 11921628 TI - Learning to choose food and drink. PMID- 11921631 TI - Long-term care. Cash-strapped Welsh homes warn of crisis. PMID- 11921632 TI - [Mathematical-technical model of passive and active vibrations of the basilar membrane]. AB - On the basis of the physiological measurements made by v. Bekesy and Johnstone, we developed a mathematical model to describe passive and active displacement patterns of the basilar membrane. Approximation of the model functions to the measured values is achieved with the aid of the linear least squares method. Using frequency mapping, the distribution of the basilar membrane displacement is presented in three-dimensional graphic form. The resulting application possibilities of this approach, for example, to electronic simulation of inner ear functions and speech processing systems, are discussed. PMID- 11921633 TI - [Experimental model of tooth mobility in the human "in vivo"]. AB - The aim of the present study was to investigate experimentally the mechanical properties of tooth deflection under external loading. These properties have a significant impact on tooth movement during orthodontic treatment. The stresses and strains caused by tooth movement influence bone remodelling, which is the basis of orthodontic treatment. The movement of a tooth as a direct reaction to the forces acting on it is termed "initial" movement. It is nonlinear and has a clearly time-dependent component. While the initial tooth movement represents the totality of the reaction mechanisms of all the tissues of the tooth unit, it is determined primarily by the mechanical properties of the periodontal ligament (PDL). The PDL is the softest tissue of the tooth unit and is therefore subject to the largest deformations when forces act on the crown of the tooth. The objective of orthodontic treatment is to achieve as precise and rapid tooth movement as possible, without provoking such undesired effects as bone and root resorption. To enable the implementation of an optimal orthodontic force system that meets these requirements, a thorough knowledge of the biomechanics of tooth movement is a must. PMID- 11921634 TI - [Standardized testing of skeletal implant surfaces with an osteoblast cell culture system. IV. Specific gene expression during differentiation]. AB - Successful osseointegration of an implant depends on the properties of the material of which it is made. A standardized cell culture system for the assessment of the biological effect of material surfaces has already been described. In the present study, this system has been extended to include the quantitative analysis of the material-dependent osteoblast gene expression. Human foetal osteoblasts (hFOB 1.19) were cultured for 3 weeks on titanium surfaces of varying roughness, and on surfaces of chromium-cobalt-molybdenum alloy (CrCoMo). Using a real time RT-PCR technique, expressions of alkaline phosphatase, collagen 1 and osteocalcin were determined as parameters of osteoblast differentiation. In comparison with CrCoMo, differentiation was accelerated on titanium. While the smooth titanium surface leads to earlier cell growth, the rough surface induces more prolonged and stronger cell proliferation. Our results confirm at the molecular level the excellent clinical biocompatibility of titanium surfaces. The real-time RT-PCR provides a new method for the quantitative assessment of material-dependent osteoblastic differentiation. PMID- 11921635 TI - Estimating the mutual information of an EEG-based Brain-Computer Interface. AB - An EEG-based Brain-Computer Interface (BCI) could be used as an additional communication channel between human thoughts and the environment. The efficacy of such a BCI depends mainly on the transmitted information rate. Shannon's communication theory was used to quantify the information rate of BCI data. For this purpose, experimental EEG data from four BCI experiments was analyzed off line. Subjects imaginated left and right hand movements during EEG recording from the sensorimotor area. Adaptive autoregressive (AAR) parameters were used as features of single trial EEG and classified with linear discriminant analysis. The intra-trial variation as well as the inter-trial variability, the signal-to noise ratio, the entropy of information, and the information rate were estimated. The entropy difference was used as a measure of the separability of two classes of EEG patterns. PMID- 11921636 TI - Automatic classification algorithms of the EEG monitor Narcotrend for routinely recorded EEG data from general anaesthesia: a validation study. AB - Impacts of hypnotic drugs on brain function are reflected in the EEG. The EEG monitor Narcotrend performs an automatic classification of the EEG using a scale which was proposed by Kugler for visual evaluation of the EEG. In this article the results of a validation study of the automatic classification algorithms implemented in the EEG monitor Narcotrend are presented. Visual and automatic classification of EEG data recorded in routine clinical practice were compared. The correlation between visual and automatic assessment was high (Spearman rank correlation r = 0.90, prediction probability Pk = 0.90) and a sufficient agreement between visual and automatic assessment was achieved for 92% of the analysed EEG epochs. The results of the study suggest that the automatic classification algorithms implemented in the EEG monitor Narcotrend yield a reliable assessment of the depth of hypnosis. PMID- 11921637 TI - Effects of two barrier creams on the diffusion of benzo[a]pyrene across human skin. AB - Barrier hand creams, often containing antiseptic agents, may provide a form of protection not only for health care professionals, but also for workers in the chemical and pharmaceutical industries. To evaluate the efficacy of two such barrier creams available on the South African market, the in vitro diffusion of a model compound, benzo[a]pyrene, through human skin at 20 degrees and 37 degrees C was studied. Treated (10 min) and untreated human skin disks (4 mm in diameter) were mounted in flow cells of a continuous flow-through diffusion apparatus. Buffer/tritiated benzo[a]pyrene was collected from the acceptor chambers at 2 hour intervals for a total of 24 hours and counted in a liquid scintillation counter. At 20 degrees C no significant differences could be detected between the flux rates of benzo[a]pyrene across barrier cream treated and untreated skin. However, at 37 degrees C Skinguard significantly increased flux rates of this carcinogen. Skin barrier creams therefore need to be carefully scrutinised with respect to their protective effects because the latter may vary for molecules with different chemical properties. PMID- 11921638 TI - Comparative cost of ART and conventional treatment within a dental school clinic. AB - BACKGROUND: The changing oral health needs in South Africa require that both the teaching and clinical techniques of atraumatic restorative treatment (ART) form a part of the restorative undergraduate curriculum. OBJECTIVE: This study was undertaken to establish and compare the estimated costing of an amalgam, composite resin and ART restoration within the Board of Health Funders (BHF) recommended scale of benefits at the School of Oral Health Sciences Oral and Dental Hospital, University of the Witwatersrand (SOHS). METHODS: Fixed and variable costs were calculated by pricing items and equipment used in each procedure. The output values were established according to the recommended scale of benefits (BHF, 1999). This enabled the calculation of contribution margins and net income for each of the three restorations. RESULTS: The annual capital cost for the ART approach is approximately 50% of the other two options (e.g. per multiple surface restoration ART = R1.58; amalgam and composite resin restorative procedures: R3.12 and R3.10 respectively), despite the fact that ART restorations are rendered in a modern dental setting. CONCLUSIONS: Our study shows that implementation of the ART approach within the clinic setting of the SOHS can be accomplished without additional cost. Furthermore ART can be performed as an economically viable alternative to conventional treatment procedures within the clinic setting. The study represents a first step towards determining the cost efficiency of implementing ART as a pragmatic and cost-effective restorative option within the SOHS, University of the Witwatersrand. PMID- 11921639 TI - Antibiotic prophylaxis for infective endocarditis: an update. PMID- 11921640 TI - Haematoma of the floor of the mouth following implant placement. AB - Placement of implants in the anterior mandibular region is generally regarded as a routine, safe procedure. This case report describes an extensive haematoma in the floor of the mouth, following such a procedure, which rapidly became life threatening, requiring an emergency tracheostomy to establish a surgical airway. The anatomic, radiographic and surgical aspects to the problem are discussed. Finally, when undertaking such procedures it is advisable to perform them reasonably close to a hospital where such a complication can be effectively and promptly handled by suitably trained persons. PMID- 11921641 TI - A review of the anatomy of the mandible. PMID- 11921642 TI - [Clinical trial of willow bark extract. We have by no means compared apples and pears]. PMID- 11921644 TI - [High blood pressure and renal lesions. Curable with hypnosis?]. PMID- 11921643 TI - [Aphthous stomatitis. What do physicians advise?]. PMID- 11921645 TI - [Complaints in the anal area. Risk a glance at the taboo zone!]. PMID- 11921646 TI - [In order to prevent anemia, osteoporosis and malignant lymphoma, sprue even with few symptoms has to be treated]. PMID- 11921647 TI - [Thyroid hormones in the treatment of iodine deficiency goiter. Superfluous like goiter?]. AB - In recent years, the widespread use of iodized salt in households and industrially produced foods has considerably improved the shortage of iodine in the German diet. Recent epidemiologic studies have shown that we are now at the threshold of adequacy. Over the last decade, views on the pathogenesis of iodine deficiency goiter have changed: while TSH was previously assumed to be the major factor underlying this form of goiter, experimental data now finger the activation of local growth factors, in particular IGF1. Iodinated organic compounds, in particular iodolacton produced by iodination of unsaturated fatty acids in the thyrocyte, are the key regulators of IGF1 activity by inhibitory action, and iodine is now the therapeutic agent of first choice for iodine deficiency goiter; long-term thyroxine treatment in TSH suppressive doses has been abandoned. The recommended daily dose of iodine is 200 micrograms; higher doses may induce or aggravate autoimmune thyroiditis, and should not be used for a length of time. In pregnant women, an adequate supply of iodine is of critical importance for optimal neurophysiological development, and the general use of iodine supplementation in pregnancy therefore remains mandatory in Germany. PMID- 11921648 TI - [Hormone replacement therapy in peri- and postmenopause. Routine use is not indicated]. AB - Hormone replacement therapy (HRT) with estrogens (in non-hysterectomized women with estrogens and progestins) during the peri- and postmenopausal period has been widely applied for many years. On the basis of new data, HRT is currently being critically reviewed. HRT administered for up to 5 years to treat climacteric hot flashes, mood changes and sleep disturbances continues to be advocated and is largely safe. When HRT is used for longer periods, as required for the prevention of osteoporosis, a possible increase in the relative risk for breast cancer must be considered. Correctly applied in combination with an adequate dose of progestins, HRT can avoid an increase in the endometrial cancer risk. HRT is no longer recommended for secondary prevention of cardiovascular disease, and its use in primary prevention has not been convincingly demonstrated. The hoped-for efficacy of HRT in the prevention of Alzheimer's disease has not been confirmed by the data. Selective estrogen receptor modulators (e.g. Raloxifene) and biphosphonates are efficacious drugs for the prevention and treatment of osteoporosis. For women at risk of developing cardiovascular disease, changes in lifestyle, lipid-lowering drugs (statins), blood pressure control, use of acetylsalicylic acid, among others, have well documented efficacy in primary and secondary prevention. PMID- 11921649 TI - [Gene, fast food and no motion. Causes of childhood obesity]. AB - Obesity in children and adolescents should be seen as a chronic disorder and not merely as a biological variant. One aspect of the pathological significance of obesity in childhood and adolescence is determined by functional and individual limitation and psychosocial impairment. Another is the fact that these children and adolescents have a higher comorbidity, and appreciably increased morbidity and mortality rates in adulthood. The major causes of the condition are lifestyle related, in particular excessive intake of energy- and fat-rich foods together with increasingly inadequate exercise. In addition, however, there also appears to be a genetic predisposition that manifests as early and massive obesity in young years. Considerable importance attaches to the recording of height and weight development during regular examinations in the doctor's office. In addition, in particular the children of overweight parents, together with their families, should be instructed and advised on the need for a healthy and appropriate diet and regular exercise. In the case of extremely overweight children, a genetic investigation would be worthwhile. PMID- 11921650 TI - [Emergencies in the practice. Acute lumbago: diagnosis and therapy]. PMID- 11921651 TI - [Against Lyme borreliosis vaccination also, soon. Until then: antibiotics as early as possible!]. PMID- 11921652 TI - [Chest pain, dyspnea, RR decrease. Suspicion of infarction led to a wrong track]. PMID- 11921653 TI - [Secondary forms of hypertension. 5. Hypertension caused by Cushing syndrome and Conn's disease]. AB - The Conn syndrome and the Cushing syndrome are major differential diagnoses in the investigation of hypertension. Primary aldosteronism (Conn syndrome) is found in < 0.5% of hypertensives. In addition to the usual laboratory investigations, magnetic resonance angiography should be performed at the start of the diagnostic work-up, since it enables a very good representation of the adrenals, and saves many diagnostic tests. Minimally invasive microsurgical techniques and the development of new drugs have greatly simplified the treatment of this form of secondary hypertension. In the case of the Cushing syndrome, which leads to hypertension in about 70% of the patients, the general practitioner should look for the typical cutaneous changes that inevitably accompany this syndrome. Only when the Cushing syndrome has been confirmed clinically and biochemically, should an imaging procedure be performed. PMID- 11921654 TI - [Cardiovascular disease screening. Ask how does is it go with sex]. PMID- 11921655 TI - [Type 2 diabetes. How can the infarction risk be reduced?]. PMID- 11921656 TI - [Mesalazine in ulcerative colitis. Fewer cases of colorectal carcinoma]. PMID- 11921657 TI - [Is it the physician's fault when type 2 diabetic patient refuses insulin? "Physicians threaten with the syringe"]. PMID- 11921658 TI - Prosthetic heart valves: why biological? AB - The replacement of heart valves only became feasible after the development of the heart-lung machine in 1953. Two groups of prosthetic heart valves were subsequently developed: biological valves that do not require anticoagulation and mechanical valves that require life-long anticoagulation with Coumadin. The incidence of heart surgery and the demographics of patients who require heart valve surgery vary worldwide; these factors influence the choice of prosthetic valve for the individual patient and are briefly reviewed. Improved biological tissue-fixation methods are also increasing the durability of biological prosthetic valves and will further favor the implantation of biological valves in the future. PMID- 11921660 TI - The influence of design on bioprosthetic valve durability. AB - Bioprosthetic heart valves have evolved over the years into remarkably useful and predictable devices. A number of specific designs have come and gone, and a few have remained. Many design changes were successful and many were not. This article describes the successes and failures of the various bioprosthetic valve designs and details the specific reasons why a particular design change succeeded or failed to improve bioprosthetic valve performance. PMID- 11921659 TI - The failure modes of biological prosthetic heart valves. AB - Bioprosthetic heart valves have been used since the 1960s, starting with the use of homograft aortic valves obtained from human cadavers. Today prosthetic heart valves are used widely, and bioprostheses account for close to 40% of all heart valve replacements. Although most bioprosthesis are still stented porcine aortic valves, the introduction of stentless valves and the increasing use of cryopreserved homograft valves has led to an upsurge of interest in bioprosthesis. There have been significant changes in the handling and fixation of porcine aortic valves; however, their modes of failure remain virtually unchanged, although many bioprosthetic valves now last for considerably longer periods. This article reviews the modes of failure of bioprosthetic heart valves. PMID- 11921661 TI - Glutaraldehyde fixation revisited. AB - Our ability to stabilize animal heart valves and to manufacture equivalent prostheses using collagenous tissues has proven to be of significant value to large number of patients over the last 30 years. Such tissue-derived bioprostheses are being gradually improved by modifications of design and slight changes in the processing of the tissues from which they originate. Nevertheless their durability does not seem to have significantly improved, and some of their drawbacks, such as their propensity to calcify, have not been eliminated. Because of this children are excluded from receiving such implants. Enhancing such structures using cells from various sources or with the aid of growth factors does not seem likely to succeed because such inert matrices are not compatible with growth and remodeling. On the other hand, stabilizing these structures by means of novel crosslinking approaches, the addition of "plasticizing" molecules, or the addition of covalently bound residues that inhibit the growth of mineral deposits, could prove to be practical ways of improving such devices. PMID- 11921663 TI - PhotoFix: unraveling the mystery. AB - The use of biological materials in prosthetic heart valves is widespread. Other articles in this issue address many key aspects of these materials. Fixation treatments, biological characteristics, mechanical performance and dysfunction, and the promise of tissue engineering are all discussed. The rest of this issue serves as an introduction to the many aspects of the use of biological materials, whether they are derived from animal sources or are bioassembled on prepared scaffolds. In this article, a particular fixation treatment, dye-mediated photooxidation (PhotoFix), is discussed. This process yields a material that is biostable, biocompatible, relatively noncalcific, and flexible. It is prepared using a relatively simple method, and the apparent chemical and physical changes in the tissue do not appear to be great. However, the resultant material exhibits dramatically altered biological properties. The mystery of why this simple process produces such dramatic changes is explored here. Despite the title of this article, like many scientific endeavors, our understanding thus far has lead to additional questions rather than providing definitive, terminal answers. Thus, "unraveling" the mystery has led to answers as well as some loose ends. PMID- 11921662 TI - Alternative fixation of bioprostheses. AB - The glutaraldehyde fixation technique for bioprosthetic tissue valves has been used for about 30 years. It is more or less generally recognized, however, that a solution to the ongoing problems of calcification and immunomodulated inflammation can only be achieved with the development of alternative fixation techniques that differ from fixation with glutaraldehyde. This review reports on the chemistry behind newly developed tissue-fixation processes and the preliminary results achieved with these. PMID- 11921664 TI - Inflammatory and immune processes: the neglected villain of bioprosthetic degeneration? AB - In an attempt to avoid the destructive process of bioprosthetic heart-valve calcification associated with the use of glutaraldehyde, valves are today prepared using low concentrations of the crosslinking reagent. In this review, we summarize our findings and those of others that confirm that the immunogenicity of such tissue is not sufficiently masked and that a defined humoral response is indeed mounted against a repertoire of antigens unrelated to those associated with vascularized and non-cross-linked xenograft organs. We demonstrate the need for increased cross-linking of tissue to satisfactorily mitigate that response; furthermore, we examine the impact of increased cross-link density on the macrophage as antigen presenting cell with respect to its involvement in both tissue erosion and pannus overgrowth. Finally we present evidence for a role of circulating antibodies in bioprosthesis calcification. PMID- 11921665 TI - Matrix metalloproteinases and tissue valve degeneration. AB - Bioprosthetic heart valves have been used as replacements for diseased heart valves for over 30 years. More than 50% of bioprosthetic valves fail within 15 years because of structural deterioration. The role of proteolytic degradation, with particular reference to the matrix metalloproteinases (MMPs) in the degeneration of aortic bioprostheses, is appraised in this minireview. It is clear that both the intrinsic and host-derived proteolytic activities present in heart-valve bioprostheses may combine with mechanical stress to bring about valve failure. PMID- 11921666 TI - The biomechanical effects of fatigue on the porcine bioprosthetic heart valve. AB - Characterization of the mechanisms of degeneration of porcine bioprosthetic heart valves (BHV) during long-term cyclic loading is required for predicting and ultimately preventing their failure. Isolation of purely mechanical effects from host biological ones is a necessary first step in understanding the fatigue process as a whole. Thus, in this review we focus on mechanical factors alone as a means of isolating their role in altering biomechanical properties and ultimately their contribution to the fatigue damage process. Mechanical evaluations included tension controlled biaxial, 3-point flexural, and uniaxial failure tests performed on cuspal tissue following 0, 50, 100, 200, and 300 x 10(6) in vitro accelerated test cycles. Overall, biaxial mechanical results indicate a decreasing radial extensibility that can be explained by stiffening of the effective collagen fiber network as well as a small decrease in the splay of the collagen fibers. Moreover, these results suggest that the loss in flexural rigidity with fatigue that we have previously measured (ASAIO 1999; 45:59-63) may not be because of loss of collagen stiffness alone, but also to fiber debonding and degradation of the amorphous extracellular matrix. We discuss the implications of these results that point toward the development of chemical treatment methods that seek to maintain the integrity of the amorphous extracellular matrix to ultimately extend BHV long-term durability. PMID- 11921667 TI - Tissue engineering of cardiac valves on the basis of PGA/PLA Co-polymers. AB - The limitations of currently used heart valve devices are well known. For prosthetic valves they include infection risk and thrombembolic complications; biologic devices have limited durability. Particularly for pediatric cardiac patients the problem of a lack of growth potential remains a serious issue. The multidisciplinary field of tissue engineering potentially offers an attractive pathway to overcome these disadvantages. The basic concept of tissue engineering is to build a new "tissue" from individual cellular components in vitro using a scaffold to provide an architecture upon which the cells can organize and develop into the desired "tissue" prior to implantation. The scaffold provides the biomechanical profile for the replacement tissue until the cells produce their own extracellular matrix. This newly generated matrix would then ultimately provide the structural integrity and biomechanical profile for the newly developed tissue structure. This work focuses on the concept of using a synthetically produced co-polymer (polyglycolic acid/polylactid acid) as the scaffold for the development of a new generation of heart valves. PMID- 11921669 TI - Prevalence of causative organisms in corneal ulcers seen at Sekuru Kaguvi Eye Unit, Harare, Zimbabwe. AB - OBJECTIVE: To investigate the prevalence of organisms causing corneal ulcers, determine frequency of ocular risk factors and assess the value of Gram stain in initial identification of pathogens. DESIGN: A descriptive cross sectional study. SETTING: Sekuru Kaguvi Eye Unit at Parirenyatwa Group of Hospitals in Harare, Zimbabwe. MATERIALS AND METHODS: We enrolled 43 patients with corneal ulcers for the study. Demographic data was obtained, measurement of visual acuity was done by Snellen's chart and a Topcon Slit Lamp was used to determine the characteristics of ulcers for all the patients. Corneal scrapings were obtained and examined by microscopy and culture methods to determine etiologic organisms. Positivity rates between Gram stain and culture methods were compared. MAIN OUTCOME MEASURES: Prevalence of different bacterial isolates in corneal ulcers, risk factors to infection and laboratory methods used for detection. RESULTS: Gram-positive cocci were the most prevalent organisms found (54%) followed by E. coli (18%). Pseudomonas, Klebsiella and Narcodia constituted about 9% each. Trauma, use of traditional eye medicine and previous viral disease were the common risk factors. CONCLUSION: Bacteria were the only organisms identified and Gram-positive cocci were the most common isolates. Gram stain of corneal smears was not reliable in initial recognition of offending pathogens. PMID- 11921668 TI - HIV-1 seroconversion incidence following pregnancy and delivery among women seronegative at recruitment in Harare, Zimbabwe. AB - OBJECTIVE: To estimate the incidence of HIV seroconversion among women following pregnancy and delivery. DESIGN: A prospective cohort of women who were HIV negative at recruitment on first antenatal care visit. MATERIALS AND METHODS: Pregnant women were invited to undergo voluntary confidential HIV counselling and blood draw for HIV testing during the first antenatal care visit as part of a prospective study of mother-to-child transmission of HIV-1. Repeat tests were conducted at delivery, six weeks post partum and at three monthly intervals until 24 months or on termination due to subsequent pregnancy, death or loss to follow up. Logistic regression modelling was used to determine independent predictors of HIV seroconversion. RESULTS: Among 372 HIV negative pregnant women who were enrolled, 66 seroconverted during follow up, resulting in a sero-incidence of 4.8 per 100 person years (95% confidence interval [CI], 3.1 to 6.5). Women who did not seroconvert during the time of pregnancy or follow up were significantly more likely to have used a condom with their partners (OR = 0.68, 95% CI = 0.47 to 0.99). Women aged 17 years and below had the highest seroconversion incidence (6.25%) followed by those aged 18 to 19 years (5.42%). Women who seroconverted and those who were HIV positive at recruitment were more likely to be married. Lack of education by the partner of a pregnant woman constituted a significant risk factor for HIV seroconversion (OR = 2.8; 95% CI = 1.1 to 11.0). CONCLUSIONS: There is a high HIV seroconversion incidence among women during pregnancy and following delivery, especially those aged 19 years and below. Being married does not protect the women from the risk of HIV seroconversion. Strategies for HIV prevention should target pregnant women and their partners. PMID- 11921670 TI - Maternal outcome in eclampsia at Harare Maternity Hospital. AB - OBJECTIVES: To study the presentation, management and determinants of maternal outcome in eclampsia at Harare Maternity Hospital (HMH) in order to design interventions for reduction of maternal mortality. DESIGN: Cross sectional descriptive study. SETTING: Harare Maternity Hospital, Harare, Zimbabwe. SUBJECTS: All women with diagnosis of eclampsia treated at HMH during an 18 month period. MAIN OUTCOME MEASURES: The study variables included age, parity, booking status, gestational age, location at time of first seizure, number of fits, seizure to delivery interval, maternal complications and the clinical management. RESULTS: There were 151 women with eclampsia from 25,425 deliveries in HMH (5.9 per 1,000 deliveries). The case fatality was 26.5%. The majority of fits (67.5%) occurred antepartum. The mothers who died were significantly older than the survivors, mean age 25.8 versus 22.3 (p = 0.007), and had a higher proportion of multiple seizures, 0.67 versus 0.39 (p = 0.009). In 38% of cases the first seizure occurred at home. The proportion of complications was higher among those who died. Deficiencies in clinical management were more common in the women who died, 39.5% versus 20.9% for survivors (OR 2.55; 95% CI 1.09 to 5.99) and they included delays in achieving delivery, inadequate clinical assessment and poor monitoring. CONCLUSION: Eclampsia remains a significant cause of maternal mortality in HMH with a high case fatality rate. Advanced maternal age, antepartum onset of convulsions and multiple fits were associated with increased risk of maternal death. There were deficiencies in the clinical management of a high proportion of cases. PMID- 11921671 TI - Caesarean section rate as a process indicator of safe motherhood programmes: the case of Midlands Province. AB - OBJECTIVE: To evaluate the Safe Motherhood Programme of Midlands Province by means of process indicators, in particular Caesarean Section Rates (CSR), in comparison with the commonly used outcome indicator Maternal Mortality Ratio (MMR). DESIGN: A cross sectional descriptive study. SETTING: Midlands Province, Zimbabwe. MAIN OUTCOME MEASURES: Process indicators of the Safe Motherhood Programme of Midlands Province, and Maternal Mortality Ratio as an outcome indicator for the nation and the province. RESULTS: For Midlands province, a population based CSR of 3.1% was calculated for 1999, which is well below the internationally recommended 5%. The figures for the eight districts ranged considerably from 0.18 to 7.1%. The provincial institutional CSR for the same year was 8.7% (range: 0.53 to 34.5) with a significantly higher rate in private institutions (24%) as compared to government run hospitals (8%), (Chi-square 398.26, p << 0.05). The Ante Natal Care (ANC) coverage ranged from 43.9 to 75.4% with a provincial average of 62.8%. The provincial institutional delivery coverage figure was 55% (range: 49.9 to 63.6%). These findings differed from the figures obtained in the Demographic and Health Survey for the same year: ANC and institutional delivery coverages for the Midlands province were 95.2% and 73% respectively. The availability of obstetric services was well above the minimum acceptable level as defined by the World Health Organisation (WHO) guidelines. CONCLUSION: MMR is not a very useful indicator to monitor progress in Safe Motherhood Programmes. The figures are unreliable, difficult to obtain if population based, and they show a wide range, even within one given year. Process indicators, especially CSR are easily accessible and give insight in the degree of unmet obstetric need and in referral patterns within one district and the province. PMID- 11921672 TI - Organophosphate poisoning and management, an update. AB - Organophosphate poisoning is characterised for the most part, by acute incidents. Management is by way of first aid (in mild poisoning) and use of atropine with or without the oximes, (in moderate to severe poisoning). Of late, it has become apparent that subchronic and chronic organophosphate poisoning are a common manifestation. This review paper summarises this triphasic nature of organophosphate poisoning. Possible future diagnostic and management techniques are also discussed. PMID- 11921673 TI - Susceptibility of Klebsiella species to quinolones and cephalosporins. PMID- 11921674 TI - Post natal care in Bubi district deserves more attention. AB - OBJECTIVES: To establish whether, where and when women in Bubi District attend Post Natal Care (PNC), the factors that influence attendance, the quality of services and traditional or cultural practices related to PNC. DESIGN: A cross sectional survey. SETTING: Bubi District, Matebeleland North Province, Zimbabwe. SUBJECTS: A multistage sample of 200 women with a child aged three to 12 months, convenience samples of 96 women with a child zero to 12 months who had attended PNC; 112 elderly women from the community and 10 nurses. MAIN OUTCOME MEASURES: PNC attendance, place and timing of PNC visit, quality of PNC, knowledge and attitudes towards PNC and traditional practices. RESULTS: 61% of the women had attended PNC. Factors associated with non-attendance were higher age and parity, home delivery and long distance between home and health facility. Almost all women attended PNC in a district health facility. Eighty three percent were seen at six weeks post partum. Forty percent of the mothers and 36.1% of the babies had received a full examination, but 37.7% and 4.9% respectively had not been examined at all. Nine of the 10 nurses interviewed were not aware of a PNC policy. Knowledge on PNC among the women in the community was poor. Some potentially beneficial and harmful traditional practices related to mother and baby were identified. CONCLUSION: PNC attendance in this study was almost three times the attendance reported through the routine National Health Information System, but lower than elsewhere in Zimbabwe. The quality of the services was rather poor and more geared towards the baby than the mother. Both health workers and the community need to better understand the importance of PNC. Because the first two weeks post partum is the period with highest morbidity and mortality, women should be advised to make a PNC visit within 14 days, or whenever they have problems, rather than at six weeks. All health facilities, including the mobile teams should be able to offer PNC. There is need to develop an 'evidence-based' PNC policy to guide nurses on what to offer to both mother and baby, also taking into consideration common traditional practices. PMID- 11921675 TI - Profile of the Nigerian sickle cell anaemia patients above 30 years of age. AB - OBJECTIVE: To examine the haematological profiles of patients with sickle cell anaemia above the age of 30 years. DESIGN: Prospective cross sectional study. SETTING: Department of Haematology University College Hospital, Ibadan, Nigeria. SUBJECTS: 98 patients with sickle cell anaemia above the age of 30 years. MAIN OUTCOME MEASURES: Clinical and haematological profiles of the patients. RESULTS: There were 75 females and 23 males. The steady state packed cell volume was 21 to 28% with a median of 24% Haemoglobin F (HbF) level in 33 patients was greater than 3% but less than 5%. Thirty six patients had a HbF which was greater than 5% but less than 10%. Twenty three patients had a HbF level which was greater than 10% but less than 15%. Five patients had a HbF level greater than 15% and only one patient had a HbF level of 22%. The severe complications of sickle cell anaemia were not observed in 87 patients (88.8%), though some age related complications like Grade 4 ischaemic necrosis of the femoral head, chronic inflammatory liver changes and ophthalmic complications were observed. Eighty six patients had never had a blood transfusion, while 76 had never been admitted into hospital. Thirty four patients had a mild to moderate degree of splenomegaly while 64 patients had mild to moderate degree of hepatomegaly. Seven of the patients had chronic ulcers which spanned five to 12 years before the study. Five patients, however, had had cerebrovascular accidents. The cohort of patients with HbF level greater than 10% had not experienced priapism, acute chest syndrome, cerebrovascular accidents or chronic leg ulcer. All the patients have had a form of Western education, have a good knowledge of the disease and possible outcome. CONCLUSION: The study strongly suggests that for the survival of the sickle cell anaemia patient, educational background of patients and their parents, awareness of hospital treatment and early presentation in hospital are major contributory factors. Also, adequate medical care, maintenance of disciplined life styles, stable or sedentary occupation and good family support, prompt and effective treatment of complications, all contribute to better health and hence a longer life span. PMID- 11921676 TI - Prenatal care utilization and foetal outcomes at Harare Maternity Hospital, Zimbabwe. AB - OBJECTIVES: To examine the association between adverse infant outcomes and maternal under utilization of prenatal care, among women delivering at Harare Maternity Hospital. DESIGN: Hospital based, cross sectional study. SETTING: Harare Maternity Hospital, Harare, Zimbabwe. SUBJECTS: A random sample of 3,864 pregnant women. MAIN OUTCOME MEASURES: Prenatal care utilization, maternal socio demographic information, as well as birth weight and other neonatal outcome characteristics. RESULTS: Of the total number of women who participated in this study 3,491 (90%) had at least one prenatal care visit. Women receiving no prenatal care, were more likely to be younger, unmarried and to have been transferred for delivery as compared with women receiving prenatal care. Women receiving no prenatal care were seven times more likely to deliver an infant weighing less than 1,500 grams, adjusted odd ratio (OR) = 7.22; 95% confidence interval (CI) 4.58 to 11.39 as compared with those who booked for care. Newborns of unbooked mothers were more likely to have a low apgar score at birth, adjusted OR = 1.71; to have been admitted to the neonatal intensive care unit, adjusted OR = 2.14, and to require intubation, adjusted OR = 3.35. A large proportion of women (31.4%) initiated prenatal care after 30 weeks gestation. CONCLUSIONS: There were significant differences between maternal characteristics and foetal outcomes in relation to booking status. Under utilization of prenatal care was associated with sub-optimal foetal outcomes. Improving the socio-economic status of women, their education and access to health care, and family planning methods are all strategies that should contribute to the reduction of adverse foetal outcomes. PMID- 11921677 TI - Voluntary testing and counselling for HIV. "Are adults in rural communities ready to test?" A descriptive survey. AB - OBJECTIVE: To describe rural adult preparedness to test for HIV, perceived susceptibility, response to peer group opinion, perceived positive and negative outcome expectations of testing. DESIGN: Cross sectional descriptive study. SETTING: Mashonaland West Province, Zimbabwe. SUBJECTS: Purposive convenience sample; 204 adults, 102 females, aged 18 to 50. MAIN OUTCOME MEASURES: Stage of preparedness to test for HIV, perceptions of susceptibility, response to peer group opinion, negative and positive expectations about testing. RESULTS: None of the 204 participants had knowingly been tested for HIV prior to the study; 55.8% had heard about Voluntary Counselling and Testing (VCT) and 21.0% had thought about testing. Of these, 15 (34.8%) had already talked to somebody about testing. When compared with married respondents, significantly more singles expressed preparedness to test for HIV in the next six months (p = 0.040). Only 14.2% of respondents expressed willingness to pay for a test. Females worried more than males about "getting HIV in the future" (p = 0.019). Singles were more likely to worry about their partners' current HIV status than married and once married respondents (p < 0.001). When compared with women, men showed more likelihood of responding to pressure from friends to test for HIV (p = 0.039). Strongly articulated fears about testing included being seen by friends at a VCT centre, stigmatisation, violence and stress. Significantly more women than men were fearful that an HIV test would be painful (p = 0.006). Similarly, women were more fearful than men about taking an HIV test (p = 0.007), the possibility of waiting for the result (p = 0.022) and returning for results (p = 0.020). Anticipated positive outcomes of testing included getting assistance in the early stage of the disease. CONCLUSION: Knowledge of HIV status acquired voluntarily in a psychologically and socially supportive environment will be a significant motivator for individuals and their partners to initiate and maintain safer sexual behaviour. Our study showed some preparedness among adults from two rural communities to test for HIV. While current strategies to promote VCT in urban areas reflect sensitivity to many of the fears of adults expressed in this study, the initiative must be expanded to embrace rural communities. There is a need to build on positive perceptions about testing and embrace a strategy that disables fear, particularly among women. The initiative must go beyond the individual as the primary target and be promoted within the broader context of the community with the assistance of credible opinion leaders. It should also link closely with other services that offer primary prevention, pilot treatment and support activities. PMID- 11921678 TI - TC99m MIBI scintigraphy in well-differentiated thyroid carcinoma. AB - OBJECTIVES: To review our experience and assess the role of TC99m MIBI thyroid scintigraphy in patients with well-differentiated thyroid carcinoma. DESIGN: Cross sectional descriptive study comparing the results of TC99m MIBI and TC99m pertechnetate scintigraphy with histology. SETTING: Ga-Rankuwa Hospital, a tertiary referral centre, MEDUNSA, South Africa. SUBJECTS: Between October 1998 and January 2000, 63 patients presenting with a single thyroid nodule were included in this study. INTERVENTION: All patients included in the study were subjected to fine needle aspiration, thyroid function tests, TC99m pertechnetate and TC99m MIBI scintigraphy. Following surgery all specimens were sent for pathological examination. MAIN OUTCOME MEASURES: The outcome of the study was measured comparing the results of pre-operative TC99m MIBI and TC99m pertechnetate scintigraphy with final histology. RESULTS: 41 of these 63 patients were found to have benign disease and 22 patients had malignant disease. In the malignant group there were 17 females and five males. The age of the patients ranged from 28 years to 85 years in the carcinoma group. Thirteen patients had follicular carcinoma, six patients had papillary carcinoma, two patients had Hurthle cell carcinoma and one patient had medullary carcinoma. TC99m MIBI correctly predicted malignancy in 21/22 patients in contrast with TC99m pertechnetate scintigraphy which diagnosed malignancy in 17/22 patients. The sensitivity of MIBI was 95% and its specificity was found to be 63%. The sensitivity and specificity of TC99m pertechnetate, however, was 77% and 29% respectively. CONCLUSION: TC99m MIBI scintigraphy can be used in association with Fine Needle Aspiration (FNA) in the pre-operative evaluation of thyroid carcinoma. PMID- 11921682 TI - The role of management in optimizing health potentials of children and adolescents. PMID- 11921680 TI - Families made by science. Arnold Gesell and the technologies of modern child adoption. AB - This essay considers the effort to transform child adoption into a modern scientific enterprise during the first half of the twentieth century via a case study of Arnold Gesell (1880-1961), a Yale developmentalist well known for his studies of child growth and the applied technologies that emerged from them: normative scales promising to measure and predict development. Scientific adoption was a central aspiration for many human scientists, helping professionals, and state regulators. They aimed to reduce the numerous hazards presumed to be inherent in adopting children, especially infants, who were not one's "own." By importing insights and techniques drawn from the world of science into the practical world of family formation, scientific adoption stood for kinship by design. This case study explores one point of intersection between the history of science and the history of social welfare and social policy, simultaneously illustrating the cultural progress and power of scientific authority and the numerous obstacles to its practical realization. PMID- 11921683 TI - [Characteristics of growth and development in children from families at social risk]. AB - INTRODUCTION: Body height and weight are important indicators of children's health status. There are many evidences that children from disadvantaged families have lower height and weight than children of the same age from families without social risk. The aim of this study was to investigate characteristics of growth and development of children from economically disadvantaged families. MATERIAL AND METHODS: The study was partly retrospective and partly prospective. The retrospective study included 509 children from disadvantaged families hospitalized at the Institute of Child and Adolescent Health Care in Novi Sad, during a five-year period. The prospective study included 90 children from disadvantaged families (experimental group) and 132 children from families without social risk (control group) hospitalized at the Institute during a six month period. Height/length, weight, head circumference, and psychomotor/intellectual development have been examined. In the retrospective study results were compared with theoretically expected values, whereas the prospective study results of experimental and control group were compared. RESULTS: In the retrospective study that included only children from disadvantaged families, 136 (26.7%) children had height/length, 173 (34%) had weight, and 86 (16.9%) children had head circumference below 10th percentile. Delay in psychomotor/intellectual development was established in 177 (34.8%) children. Children from families with social risk have significantly more often height/length, weight, head circumference and developmental delay than theoretically expected. In the prospective study 40 (44.4%) children from experimental group had height/length, 29 (32.2%) had weight, 20 (22.2%) children had head circumference below 10th percentile, and 17 (26.2%) had delay in psychomotor/intellectual development. Children from disadvantaged families (experimental group) significantly more often had delay in growth and development comparing with children from families without social risk (control group). CONCLUSION: Children from disadvantaged families significantly more often exhibit delay in growth and development, comparing with children of the same age from families without social risk. Therefore, pediatricians should consider social risk factors whenever treating children with growth or developmental delay. PMID- 11921684 TI - A simple and reliable step-test for indirect evaluation of aerobic capacity. AB - Substantial data confirm the benefits of physical activity, but it is necessary to prescribe exercise correctly in regard to aerobic capacity of each patient. When undertaking investigation of a great number of people, it is necessary to apply a simple, fast, reliable and inexpensive test. The aim of this study was to evaluate the validity of a new step-test in physically active subjects. Thirty eight male athletes (mean age 16.6 +/- 0.3 years) and 38 nonathletes of the same age, underwent Astrand cycle-ergometer test and a new ergometric step-test. In the ergometric step-test a fixed workload (W) of 12 kJ was used, whereas the bench height (h in cm) was determined according to body mass of each subject (h = W/2.g.n) (W = workload in kJ; BM = body mass in kg; n = step rate, 25 steps + min 1). The step-test lasted 6 minutes. Values of maximal oxygen consumption (VO2 max) were determined using a nomogram, according to workload and heart rate (HR). Comparing data concerning VO2 max in Astrand and step-test, no significant differences were observed in either group: athletes (3505.6 +/- 784.7 vs. 3294.9 +/- 653.4 mL.min-1 for the step-test; p > 0.05) and in nonathletes (2563.3 +/- 469.3 vs. 2471.8 +/- 353.6 mL + min-1; p > 0.05). In our opinion our new, modified ergometric step-test is a test with high degree of validity and can be used as an easy and fast procedure instead of Astrand test which is most frequently used for indirect evaluation of aerobic capacity. PMID- 11921685 TI - [Work with parents of children with chronic illness or developmental disability]. AB - INTRODUCTION: It is more and more important to work with parents of disabled children or children with chronic illness. PURPOSE: The goal of this study was to investigate mother's presence during hospitalization in regard to the child's handicap. We also wanted to establish if parents were trained to work with children with developmental disabilities. MATERIAL AND METHODS: In order to learn if parents of disables or chronically ill children were trained to help them, parents of 99 children hospitalized at Principovac Hospital were asked to fill out a questionnaire. RESULTS AND DISCUSSION: It was established that parents of children with multiple disabilities were best trained, whereas parents of mentally handicapped children were the least trained. Parents of physically handicapped children (63.6%) and children with multiple disability (63%) were mostly present during hospitalization. PMID- 11921687 TI - [Otoacoustic emissions--a step closer to understanding cochlear function]. AB - INTRODUCTION: Otoacoustic emission is a phenomenon that may occur in a large group of normal-hearing subjects. Ear is an organ which collects and treats sounds from the surrounding environment, and nerve fibres transmit the perceived information to the brain in terms of nerve signals. However, ear can also produce a sound (otoacoustic-emission), which can be recorded using very sensitive microphones placed in the ear canal. OTOACOUSTIC EMISSION: Otoacoustic emissions may be spontaneous (SOAE) or may be a response to given auditory stimuli: transiently evoked otoacoustic emissions (TEOAE) and distortion product otoacoustic emissions (DPOAE). Applications of otoacoustic emissions in clinical and experimental medicine are various. Hearing screening program of newborns aims to identify infants with permanent bilateral hearing loss in the first days after birth. Screening programs are based on recordings of transiently evoked otoacoustic emissions (TEOAE) in the first step, and then (if it is necessary) brainstem evoked response audiometry. Transiently evoked otoacoustic emissions may also be used for differential diagnosis of cochlear and retrocochlear disorder. UTILIZATION: In cases of substantial retrocochlear pathological findings without cochlear involvement, transiently evoked otoacoustic emissions should be recordable because outer hair cell function is unaffected. It is possible to detect ototoxic effects of drugs before hearing loss can be detected by other methods (pure-tone audiometry). In a similar way, using otoacoustic emissions, it is possible to detect noise induced damages of the outer hair cells. Otoacoustic emissions provide a unique means for assessing the integrity of the medial efferent pathway. Further research into the properties of various types of otoacoustic emissions would contribute to understanding of cochlear function, and in particular, the relative roles of inner and outer hair cells. Evoked otoacoustic emissions are directly related to outer hair cell function and may be somewhat independent of inner hair cells. At present, there is insufficient evidence to indicate whether inner and outer hair cell disorders can exist separately, and if they can, whether they are so strongly correlated that statistically the distinction is meaningless. Applications of otoacoustic emissions in clinical practice is an important route in furthering our understanding. PMID- 11921686 TI - [Treatment of extreme obesity with a very low calorie diet]. AB - INTRODUCTION: This paper presents the results of treatment of very obese persons with a very-low-calorie diet (VLCD). MATERIAL AND METHODS: A group of 28 extremely obese subjects, average age 32 years, was treated in the course of one month by the following regime: 3-4 l of mineral water with "Enemon" pulv. 3 x 1 and vitamin substitution and allopurinol 300 mg/day. RESULTS: In addition to a statistically significant (p < 0.5) loss of body mass, significant changes were observed in regard to decrease of atherogenic lipid profile (cholesterol, triglycerides, LDL-cholesterol, apoprotein B), as well as lowered level of hyperinsulinism which, though, was not statistically significant. The degree of protein catabolism did not reach statistical significance, and the results were also more favourable with respect to potential changes in electrolytes, as well as with respect to the degree of hyperuricemia in regard to subjects being under the regime of total starvation. DISCUSSION AND CONCLUSIONS: VLCD represents an extremely efficient and safe therapeutic procedure which, apart from body mass loss, is characterized by favourable changes in metabolism of lipids and a decrease in hyperinsulinism, which eventually results in reduction of the risk from early and accelerated atherosclerosis. PMID- 11921688 TI - [Occurrence of sexually transmitted diseases in Vojvodina during the last 20 years]. AB - INTRODUCTION: Sexually transmitted diseases are the most often registered communicable diseases in a great number of countries. The aim of this study was to analyze dynamics and distribution of gonorrhea, syphilis and scabies in Vojvodina region during the last twenty years. MATERIAL AND METHODS: Epidemiological characteristics of gonorrhea, syphilis and scabies were analyzed on the basis of data obtained from the Section of Epidemiology of the Institute of Public Health in Novi Sad. The research included the period between 1980 and 1999, with sex and age distribution of patients. Morbidity rates were given per number of inhabitants of Vojvodina. RESULTS: In the period between 1980-1999 there were 454 registered patients with the diagnosis of syphilis in Vojvodina. The morbidity ratio was highest in 1980 (3.41/100.000), and lowest in 1991 (0.24/100.000). In the twenty-year period there were more patients with gonorrhea, than patients with syphilis. There were 44.621 registered patients with gonorrhea. The maximum morbidity ratio was in 1980 (25.09/100.000), but the minimum was in 1998 (1.68/100.000). Within the examined period scabies was recorded in 56.490 patients. The highest morbidity ratio was in 1984 (232.37/100.000) and the lowest was in 1992 (73.56/100.000). DISCUSSION: The average morbidity ratio of syphilis in USA, between 1992-1994, was 11.8/100.000 and at the same time in Vojvodina it was only 0.42/100.000. In Vojvodina most patients with this disease were 20-39 years old. In the same period in USA the ratio of gonorrhea patients was 309/100.000 and in Vojvodina it was 2/100.000. In this group also, most patients were 20-39 years old. However, scabies mostly appeared at the age of 7-14. CONCLUSION: According to the obtained results, the number of registered patients with these three diseases in Vojvodina stagnates or it decreases. In order to deal with real data, it is necessary to report these diseases regularly. PMID- 11921690 TI - [The role of Professor Stanoje Stefanovic in the organization of education at the Medical School in Novi Sad and his contribution to the development of internal medicine--on the 10th anniversary of his death (1911-1991)]. AB - This paper presents the role of Academician Stanoje Stefanovic in organization of teaching activities in internal medicine and physiopathology at the Faculty of Medicine in Novi Sad as well as his contribution to development of internal medicine in Novi Sad and Province of Vojvodine. It also deals with chronological evolution of the Department of Internal Medicine of the General Provincial Hospital into a modern Clinic of Internal Diseases. PMID- 11921689 TI - [Use of aminopenicillins in hospitals and outpatient facilities]. AB - INTRODUCTION: Ampicillins are thought to be the most widely used antibiotics. Although ampicillin is included in the WHO list of essential drugs, its per-oral administration is not recommended, whereas amoxicillin is recommended due to its identical antimicrobial spectrum and better pharmacokinetic characteristics. Instead, per oral form of amoxicillin is recommended because of its identical antimicrobial spectrum and better pharmacokinetic characteristics. MATERIAL AND METHODS: Drug utilization evaluation of aminopenicillins (ampicillin and amoxcillin) was performed at the Clinic for Infectious Diseases. Clinical Center of Novi Sad and in the Outpatient Health Care Center, Novi Sad "Liman" (Department of General Practice Children's Health Care Department and Ear, Nose and Throat Department) during one calendar month in the year 2000. RESULTS: The use of aminopenicillin was 5.1 Defined Daily Doses per 100 bed-days (5.1 DDD/100 BD) at the Clinic for Infectious Diseases. Ampicillin was used in 37.64% of patients and amoxicillin in 13.13%. In Outpatient Health Care Center (OHCC), Department of General Practice, aminopenicillins were prescribed in 1.02 Defined Daily Doses per 1000 inhabitants per day (DDD/1000 inh/day); ampicillin being prescribed in 24.51% and amoxicillin in 75.49%, and it is an optimal ratio. In Children's Health Care Department, OHCC, in children older than 14 years, aminopenicillins were prescribed in 0.37 pediatric DDD/1000 inh/day (ampicillin 40.5%, amoxicillin 59.5%). In the same department, in the age group between 3 and 14 years it was 0.31 pediatric DDD/1000 inh/day (ampicillin 22.5%, amoxicillin 77.5%), while in the age group from 1 to 3 years there were 0.006 pediatric DDD/1000 inh/day prescribed (amoxicillin 100%). At the Ear, Nose and Throat Department, OHCC aminopenicillins were prescribed in 0.02 DDD/1000 inh/day (amoxicillin 100%). CONCLUSIONS: Our research showed a change in habits in regard to prescriptions towards the recommended protocol. Oral administration of ampicillin has decreased, especially in comparison to the earlier period in which it had dominated in our primary health care. PMID- 11921691 TI - [Validation of the Alvarado score in the diagnosis of acute appendicitis]. AB - AIM: The aim of the study was to find out the values of the Alvarado score in diagnosis of acute appendicitis and its clinical applicability in our conditions. MATERIAL AND METHODS: A prospective study performed from November 15 to January 20, 2001 included 68 patients of whom 52 were operated. Patients were examined at the emergency department, and all received Alvarado score, whereas the surgeon made the decision if there were indications for surgery. In order to verify the diagnosis, we also used histopathological findings. RESULTS: From 52 operated patients, 43 (82.7%) had Alvarado score 7 or more, which confirms the diagnosis of acute appendicitis, and in all these patients the diagnosis was proved both intraoperatively and histopathologically. From 9 (17.2%) operated patients that had score less than 7, diagnosis of chronic appendicitis was proved in 3; in the rest 6 acute appendicitis was established. Alvarado score has high sensitivity of 87%, and average specificity of 60%. CONCLUSION: In diagnosis of acute appendicitis Alvarado score has a high diagnostic value (82.7%). Alvarado score is a noninvasive, safe diagnostic procedure, which is simple, fast, reliable and repeatable, it can be used in all conditions, without expensive and complicated supportive diagnostic methods. Alvarado score increases the diagnostic certainty of clinical examination in diagnosis of acute appendicitis. PMID- 11921692 TI - Comparison of the fat elimination between long-chain triglycerides and medium chain triglycerides in rats with ischemic acute renal failure. AB - To guide the administration of fat emulsion in the nutritional support of acute renal failure (ARF), pharmacokinetic analysis with an one-compartment open model after bolus intravenous injection was performed to compare the elimination kinetics of long-chain triglycerides (LCT) and medium-chain triglycerides (MCT) in ischemic acute renal failure rats. Sprague-Dawley rats were randomized into four groups, namely LCT normal group, LCT ARF group, MCT normal group and MCT ARF group. The model of ischemic acute renal failure was induced by clamping the left renal artery for 60 min and contralateral nephrectomy. All the rats were fasted with water ad libitum for 10 h before 0.3 g/kg body weight of 10% Intralipid (LCT) or 10% Lipofundin (MCT: LCT = 50:50) was injected as a bolus to them via the tail vein. The serum triglyceride concentration was determined at 2, 10, 40, 70, 100, 130 and 160 min after intravenous injection for kinetic analysis. The results showed that the elimination rate constant (ke) of LCT ARF group was significantly decreased, while the half life period (t1/2) of it was significantly longer than those of LCT normal group. The ke and t1/2 of MCT showed no statistical difference between normal and ARF groups. In the normal group the ke of LCT was significantly decreased compared with MCT whereas the t1/2 was significantly prolonged. In the ARF group the ke of LCT was much less than that of MCT while the t1/2 was much longer. The serum insulin levels of both MCT groups were significantly higher than those of LCT groups. These results indicate that MCT will be eliminated more rapidly than LCT in ARF rats. MCT may also increase the secretion of insulin. In conclusion, MCT may be more favorable than LCT in the nutritional management of ARF. PMID- 11921693 TI - Acute renal failure after thrombotic agent therapy due to excessive bleeding after cosmetic liposuction. AB - Suction lipoplasty is practiced popularly for obesity control and cosmetic purposes and severe complications are rare. We report the first documented case of acute renal failure, with anuric period, that developed after excessive bleeding following cosmetic liposuction. The use of homocoagulase and the presence of rhabdomyolysis might have contributed to the acute renal failure and disseminated intravascular coagulation observed in this case. PMID- 11921694 TI - The risk of radiocontrast nephropathy in patients with cirrhosis. AB - A retrospective case-control study was conducted to determine whether liver cirrhosis might be a risk factor for radiocontrast induced nephropathy as has been suggested. Data from 72 patients with cirrhosis and 72 patients without cirrhosis who all received 100-150 ml of low osmolality radiocontrast medium for abdominal or chest computerized tomography scan were reviewed. Blood urea nitrogen and creatinine were recorded before and 48-72 h after the administration of an intravenous radiocontrast agent. Acute renal failure developed in two patients with cirrhosis (2.8%) and one patient in the control group (1.4%). This difference was not significant. There was no significant change in blood urea nitrogen and creatinine in either group after radiocontrast injection. Both of the cirrhotic patients who developed radiocontrast induced nephropathy had received high-dose diuretics and were hypovolemic. We conclude that hepatic cirrhosis per se may not be a risk factor for radiocontrast-induced nephropathy. PMID- 11921695 TI - Ionized serum calcium levels during acute renal failure: intermittent hemodialysis vs. Continuous hemodiafiltration. AB - BACKGROUND: Achieving "adequacy of dialysis" includes the maintenance of normal serum ionized calcium concentrations and is an important therapeutic goal in the treatment of acute renal failure (ARF). It is unknown whether this goal is best achieved with intermittent or continuous renal replacement therapy. METHODS: We compared the effects of continuous veno-venous hemodiafiltration (CVVHDF) and intermittent hemodialysis (IHD) on serum ionized calcium concentrations using daily morning blood tests in 88 consecutive intensive care patients of which half were treated with IHD and half with CRRT. RESULTS: Mean patient age was 54 +/- 14 years for IHD and 60 +/- 14 years for CVVHDF (NS). However, patients who received CVVHDF were significantly more critically ill (mean APACHE II scores: 24.4 +/- 5.1 for IHD vs. 29.2 +/- 5.7 for CVVHDF, p < 0.003). Before treatment, the mean ionized calcium concentration was 1.177 +/- 0.03 mmol/l for IHD and 1.172 +/- 0.04 mmol/l for CVVHDF (NS), with abnormal values in 51.6% of IHD patients and in 68% of CVVHDF patients (NS). During treatment, hypocalcemia was significantly more common among CVVHDF patients (24.5% vs. 14.9%; p < 0.011) while hypercalcemia was more frequent during IHD (36.1% vs. 25.6%; p < 0.019). CONCLUSIONS: Abnormal serum ionized calcium concentrations are frequent in ARF patients before and during renal replacement. Once dialytic therapy is applied, CVVHDF is more likely to lower serum calcium concentrations, while IHD is more likely to induce hypercalcemia. Appreciation of these different biochemical effects may assist clinicians in adjusting dialytic therapy in selected patients. PMID- 11921696 TI - Relative erythrocytosis of patients with end stage renal failure undergoing CAPD. AB - BACKGROUND AND AIM: Although the majority of patients with end stage renal failure have anemia, some have relative erythrocytosis. Patients treated with continuous ambulatory peritoneal dialysis (CAPD) having relative erythrocytosis were studied in order to determine the factors that would be responsible. METHODS: Nine out of 89 CAPD patients (10%) were identified as having relative erythrocytosis. Age-, sex- and duration of disease-matched eight patients undergoing CAPD were taken as control. Beside factors of etiologies of renal failure, smoking, renal cysts, viral hepatitides, residual renal function, the adequacy of CAPD, nutritional status, hypertension, serum levels of erythropoietin, IL-1, IL-6, TNF-, and IGF-1 levels were also investigated. RESULTS: Relative erythrocytosis occurred most often in diabetic and amyloidosis patients. None of the parameters studied were found to be significantly different between groups. During 2-year follow-up, although statistically non-significant, patients having relative erythrocytosis seemed to have higher mortality rate due to vascular complications. CONCLUSION: No single factor seemed to explain erythrocytosis in patients undergoing CAPD. Being diabetic or with amyloidosis may increase the risk. PMID- 11921697 TI - The association of interdialytic weight gain with nutritional parameters and mortality risk in hemodialysis patients. AB - High interdialytic weight gain (IDWG) is considered as an indicator of noncompliance but could also be interpreted as an index of appetite. This study was designed to investigate the relationship IDWG with malnutrition and mortality risk in hemodialysis (HD) patients through a follow-up of 24 months. We divided HD patients into two groups according to their IDWG as Group I (IDWG < 3% of dry weight/day) (27 patients; age 46.8 +/- 21.1 years; HD duration: 28.3 +/- 39.5 months) and Group II (IDWG > or = 3% of dry weight/day) (41 patients; age 40.9 +/ 11.3 years; HD duration: 54.7 +/- 38.7 months). We investigated malnutrition through biochemical analysis (albumin, prealbumin, total cholesterol, creatinine, predialysis potassium and phosphorus levels), normalized protein catabolic rate (nPCR), anthropometric measurements. On initial assessment, group I had significantly lower predialysis creatinine, prealbumin and potassium levels than Group II (p < 0.0001, p < 0.01 and p < 0.001, respectively). At the 24th month, there were significantly lower creatinine, prealbumin, potassium and phosphorus levels in the low-IDWG group. Group I had significantly lower nPCR, body weight, body mass index and triceps skinfold thickness during the follow-up. Over the 24 months, 13 (48.1%) Group I patients and nine (21.9%) Group II patients exhibited loss of dry weight (p < 0.02). According to the survival curves prognosis was significantly poorer for Group I than Group II (2-year survival 74.0% and 92.6%, p < 0.03). Group I individuals with low albumin levels had the worst survival rate (57.1%). In conclusion there is a strong association of IDWG with nutritional parameters in HD patients. Our study draws attention for a possible risk of developing malnutrition in a HD patient with low IDWG. PMID- 11921699 TI - Development of chronic renal failure in adult pure mesangial glomerulonephritis. AB - Mesangial glomerulonephritis is uncommon histopathological pattern as a part of primary glomerular diseases. 64 (6.6%) patients of 968 non-transplant renal biopsies done at our department presented pure mesangial glomerulonephritis, without evidence of IgA nephropathy or systemic disorders. 22/64 (34.7%) of the patients presented with nephrotic syndrome, 12 were male, 10 female, aged 33.32 +/- 3.14. All of them were treated with steroids, 3/22 revealed complete remission, 4/22 partial remission and 6/22 did not respond to steroids, and later combined therapy with cyclophosphamide and cyclosporine. They developed end-stage renal failure within 4-8 years. 42/64 (65.3%) patients, 26 male, 16 female, aged 32.29 +/- 1.74 presented with erythruria and non-nephrotic proteinuria. Because of the absence of nephrotic syndrome only anti-hypertensive treatment was performed in this group of patients if necessary. One patient died because of hypertension complication (intracranial hemorrhage), two developed end-stage renal failure follow-up of 12 and 6 years. PMID- 11921698 TI - Effects of hormonal replacement therapy on oxidative stress and total antioxidant capacity in postmenopausal hemodialysis patients. AB - BACKGROUND: Oxidative stress is known to be implicated in the pathogenesis of atherosclerotic cardiovascular disease. Many studies have demonstrated that hormone replacement therapy (HRT) has beneficial effects on oxidation injury in postmenopausal women with normal renal function. In this study, we examined the effects of HRT on plasma malondialdehyde (MDA) level and total antioxidant capacity (TAC) in postmenopausal hemodialysis women. METHODS: We randomly assigned 70 postmenopausal women on maintenance hemodialysis into either a HRT group or a control group. Oral conjugated estrogen (0.625 mg) combined with medroxyprogesterone acetate (2.5 mg) was given daily for 12 weeks in HRT group. Plasma MDA, TAC, albumin, uric acid and C-reactive protein (CRP) were measured before, 4 and 12 weeks after the start of medication in the HRT group. In the control group, the same parameters were measured without HRT. RESULTS: There was no difference in baseline values between the two groups. In the control group (n = 32), all these parameters showed no change at 4 and 12 weeks. HRT decreased MDA from 1.32 (0.55-1.88) microM to 1.08 (0.44-1.50) microM (p < 0.001) at 4 weeks and to 1.11 (0.50-1.37) microM (p < 0.001) at 12 weeks (n = 33). TAC was not changed at 4 weeks, however, it decreased from 1.59 (1.27-2.00) mM to 1.45 (1.08 1.65) mM (p < 0.05) at 12 weeks. The albumin, uric acid and CRP levels were not changed significantly after HRT. CONCLUSIONS: These results suggest that HRT has a favorable effect on oxidative stress in postmenopausal women with ESRD as in the general population. PMID- 11921700 TI - Increased urinary excretion of N-acetylglucosaminidase in subjects with impaired glucose tolerance. AB - N-acetylglucosaminidase (NAG) is a lysosomal enzyme produced by renal proximal tubular cells and has been widely used as a marker, which indicates a degree of renal tubular damage. An increase in urinary NAG excretion is though to result from the renal tubular damage. The aim of this study was to evaluate whether even mild hyperglycemia causes an increase in urinary excretion of NAG, which is a renal tubular protein. We examined urinary NAG excretion in overnight urine in 22 Japanese men with impaired glucose tolerance (IGT) for more than two years (IGT group) and 41 healthy control subjects matched in age, gender, BMI and blood pressure (control group). Urinary NAG excretion levels of IGT group and control group were 2.89 (1.23-7.97) and 2.22 (0.60-4.93) U/g creatinine, median (range), respectively. The IGT group showed significantly higher urinary excretion of NAG compared to the control group (p < 0.01). Several studies have indicated that plasma proteins filtered through the glomerular capillary may have intrinsic renal toxicity. Recently, we have reported that urinary excretion of plasma proteins (ceruloplasmin, IgG4 and IgG) with molecular radii of approximately 45 55 A is increased in subjects with IGT compared to healthy control subjects with normal glucose tolerance. Considering the present result together with our recent finding, we suggest that increased urinary excretion of NAG in the mildly hyperglycemic subjects may be due to the adverse effects of the plasma proteins highly filtered through the glomerular capillary on tubular cells. PMID- 11921701 TI - Epoetin treatment improves red blood cell and plasma antioxidant capacity in hemodialysis patients. AB - The efficiency of human recombinant epoetin in alleviating anemia in hemodialyzed patients has been well documented. However, the effects of rhEPO therapy in correction of antioxidant capacity are not completely explained. In this study we examined both extracellular (plasma) and intracellular (red blood cells) antioxidant potential in hemodialyzed patients before and after three and six months of epoetin treatment by evaluating markers of oxidative stress (malondialdehyde) and antioxidant capacity (thiol groups, superoxide dismutase, and glutathione peroxidase). Six months of treatment with epoetin was followed by significant increases in thiol groups, superoxide dismutase and glutathione peroxidase activities in both plasma and red blood cells of hemodialyzed patients. Hence, during accelerated erythropoiesis, an increase in the number of young hematopoietic cells may replenish erythrocyte superoxide dismutase and glutathione peroxidase activity. However, the consequences of an imbalance between enzymatic antioxidant system (higher superoxide dismutase and lower glutathione peroxidase activity) that exists in these patients are the very high red blood cell and plasma malondialdehyde levels. These results suggest that, in spite of epoetin treatment and improvement in red blood cells and plasma antioxidant capacity, the production of reactive oxygen species overwhelms the intracellular and extracellular antioxidant capacity. PMID- 11921702 TI - Down-regulation of CD43 molecule expression on intraperitoneal neutrophils in CAPD patients with peritonitis. AB - To assess the release of proteases from neutrophils infiltrated into the peritoneal cavity in continuous ambulatory peritoneal dialysis (CAPD), we investigated the regulation of CD43, LAM-1 and Mac-1 expression on the neutrophil plasma membrane using FACS analysis in CAPD patients with peritonitis. Five CAPD patients with peritonitis and five CAPD patients without peritonitis were studied. CD43 expression was immunohistochemically determined in both groups of patients using flow cytometry, and comparisons were made between the two groups. Down-regulation of CD43 and LAM-1, and up-regulation of Mac-1 were demonstrated on neutrophils obtained from CAPD dialysate of peritonitis patients after 1-h dwell time. Further up-regulation of Mac-1 developed until a dwell time of 4 h. Immunoblot analysis for neutrophil lysate from dialysate showed the presence of the asialo form of CD43 molecules and their fragments, which may be produced by cleavage of the CD43 molecule at extracellular sites. The intraperitoneal neutrophils in dialysate from CAPD patients with peritonitis are continuously activated during dwell time, and proteases may be released from neutrophils into dialysate after only a short dwell time. PMID- 11921703 TI - Acute renal failure associated with nonfulminant hepatitis A virus infection. AB - Hepatitis A is usually a mild self-limiting infection of the liver. Nonfulminant acute renal failure very rarely complicates type A viral hepatitis. An unusual case, a 32-year-old female with serologically proven acute hepatitis A infection, was complicated by acute renal failure and the patient in this study is the first case associated with Cushing's disease. She recovered, and the laboratory tests returned normal one month after initial hospitalization. Although the mechanism responsible for renal failure in acute hepatitis A virus infection is still uncertain, possible causes are discussed with the review of literature. PMID- 11921704 TI - [Genomic approach to allergic diseases: present and future perspectives]. AB - The rapid advances in genomic research have a major impact on biomedical sciences. In this review the authors summarize the current results of the genomic investigation of allergic diseases. From a genetic point of view allergy is multifactorial, which means that the susceptibility to the disease is determined by interactions between multiple genes, and involve important nongenetic factors such as the environment for their expression. There are two widespread methods for searching for disease susceptibility genes in allergy: (1) genome-wide search and (2) candidate gene association studies. Until now five groups have completed genome-wide searches for asthma, and almost 500 genetic linkage and association studies have reported more than 500 atopy and asthma loci throughout the genome. In this review we selected those results, which were consistently confirmed by several independent studies, or appeared particularly important, or interesting. On the basis of this, 9 chromosome regions can be associated with the susceptibility to allergic diseases: 2q, 5q31-q33, 6p21.3, 7q31, 11q13, 12q14.3 q24.31, 14q11.2-q13, 16p21, 17q11.2. According to the results of the human genome programs, and association studies, the authors discuss the possible roles of candidate genes found in these loci in the pathomechanism of allergy. Besides, the authors summarize briefly the results of pharmacogenomics, and animal linkage and genetics studies related to allergy. PMID- 11921706 TI - [Perioperative temporary pacemaker therapy]. AB - INTRODUCTION: Numerous factor can precipitate progression of incomplete heart block to complete heart block in the perioperative period. While there is a consensus of the indications of acute temporary pacing, there is no widely accepted guidelines for the elective perioperative temporary pacing. The indications are clear in the cases of second or third degree atrio-ventricular block, bradycardias, bradyarrhythmias with frequency less than 50/min not responding to atropine, but there are many debate in the cases of first degree atrio-ventricular block with bifascicular or left bundle branch block. Furthermore, during the operation bradyarrhythmia, complete atrio-ventricular block not responding to atropine can develop without any sign on the previous ECG at rest. METHODS AND RESULTS: The authors review the different methods of pacing (transvenous, transcutaneous, transoesophageal), summarize the advantages and disadvantages of each method, the complications and their prevention. Patients' condition, concomitant medical treatment, designed procedures and staffs' experience in the introduction of pacemaker electrode must be considered in questioned cases. CONCLUSIONS: The transcutaneous method is easily performed, don't need a lot of experience. It must be available in every operating theatre and in the questioned cases may be enough until the introduction of the transvenous electrode, if the field of operation make the electrode placement possible. PMID- 11921705 TI - [Role of tetrahydrobiopterin in the regulation of activity of human placental nitric oxide synthase in normal and pre-eclamptic pregnancies]. AB - The discovery of nitrogen monoxide (NO), the structurally simplest signal molecule has motivated intensive research work in medical biochemistry. Among the enzymes synthesizing NO, the type III isoform (or endothelial NO-synthase) is produced by the syncytiotrophoblasts of human placenta and the endothelial cells of placental and umbilical blood vessels. In this paper some of the characteristic molecular properties and certain evident or putative physiological roles of NO-synthase III (NOS III) as well as those experimental results of the author's group that demonstrate the concept of dual regulation of placental NOS III activity by Ca2+ and tetrahydrobiopterin (BH4) are reviewed. BH4 promotes the formation of the enzymatically active stable homodimeric structure of NOS III. In addition, BH4 remains bound to the enzyme during the catalytic cycle and takes part in the electron transfer necessary for NO synthesis. Work of the author's group and data from the literature indicate that BH4 is bound tightly to one of the subunits of the homodimeric enzyme, whereas the binding affinity to the second binding site (which is present on the second subunit) is markedly smaller. Tightly and loosely bound BH4 molecules are supposed to ensure the basal and the BH4-regulated enzyme activities, respectively. In first trimester placentae NOS III exhibits 42-81% of the BH4-dependent maximal activity, whereas in terminal placentae this value falls to 16-59%. BH4 concentrations of individual placentae are variable and as a rule, it decreases during pregnancy together with a decrease of enzyme concentration. Studies of the author's group indicate that in some of the preeclamptic pregnant women, activation of placental NOS III by physiological concentrations of BH4 is not detectable while the basal activity of the enzyme remains unchanged. Mechanisms that can explain how the malfunction of placental NOS III might be involved in the pathogenesis of preeclampsia are discussed. PMID- 11921708 TI - [The world of Doctor Bulgakov]. PMID- 11921707 TI - [Immunohistochemical study of P-cadherin in breast cancer]. AB - INTRODUCTION: Cell adhesion molecules play a significant role in the cellular connection of normal cells. The cadherins are believed to act as tumour suppressors, and their altered expression and function have been associated with tumour development. AIM AND METHODS: The authors examined the expression of a Ca++ dependent intercellular adhesion molecule, P-cadherin using an immunohistochemical method in 69 surgically resected breast carcinomas. RESULTS: P-cadherin was detected in 30 cases (43.5%, cytoplasmic and/or membrane staining). The expression of P-cadherin was independent of tumour size and lymph node status, but correlated with a high tumour grade (grade III). In contrast, expression of E-cadherin correlated with lower tumour grade (grade I-II). P cadherin expression was not detected in invasive lobular carcinomas. CONCLUSION: In general, P-cadherin was expressed at a lower frequency compared to E-cadherin, alpha-, and beta-catenin. These results suggest that an inverse relationship may exist between E- and P-cadherin in relation to grade, and that the expression of P-cadherin may be a marker of aggressiveness. PMID- 11921709 TI - [Educational poetry of Antal Pucz about the blood circulation from 1790]. PMID- 11921710 TI - Using denture cleansers safely. PMID- 11921711 TI - Getting the lowdown on back pain. PMID- 11921712 TI - Postoperative peril. A nurse's back surgery goes tragically wrong. PMID- 11921713 TI - Ebola: preparing for the worst. PMID- 11921715 TI - Handling verbal orders safely. PMID- 11921714 TI - Taking HAART. How to support patients with HIV/AIDS. PMID- 11921716 TI - Sharing. Nothing of value. Sometimes, life's most precious gifts don't cost a thing. PMID- 11921717 TI - Photo guide. Focus on autotransfusion. PMID- 11921718 TI - Truth telling in patient care. Resolving ethical issues. PMID- 11921719 TI - Detecting dehydration and malnutrition in the elderly. PMID- 11921720 TI - Unraveling the mystery of acute pancreatitis. PMID- 11921721 TI - Morally managing executive mistakes. AB - Medical errors have been the subject of extensive discussion for many years. In contrast, management mistakes have not received the same scrutiny. Why is this true, and what are some of the factors contributing to management mistakes? What constitutes a mistake or error? How do mistakes in management compare with those in medicine? When and how should mistakes be disclosed? What are appropriate options for dealing with them productively and ethically? How can the incidence of mistakes be reduced? This article is intended to stimulate discussion about a critical topic--one that has received inadequate attention by both healthcare administration and the field of organizational ethics--with important implications for improving executive and organizational performance. PMID- 11921722 TI - Why pursuing mistakes in management is a mistake. PMID- 11921723 TI - Protecting the community's trust: coping with executive mistakes. PMID- 11921724 TI - Preventive ethics and cultivating integrity. PMID- 11921726 TI - Farmers face a big stinking mess. PMID- 11921727 TI - [25-anniversary of the Institute of Infectious and Parasitic Diseases, Medical Academy in Warsaw (1976-2001)]. PMID- 11921728 TI - [Infectious mononucleosis]. AB - The diagnosis of infectious mononucleosis is usually based on the clinical signs and symptoms, characteristic changes in the peripheral blood picture and the results of specific and non-specific serologic tests. However, despite of these precise diagnostic criteria this disease is often misdiagnosed, misunderstood and therefore treated in a wrong way. Therefore, because of our long lasting clinical experience based on a very large number of cases, we have decided to present the results of our studies on the clinical features of this interesting disease as well as some laboratory findings (serum bilirubin level and AspAt, AlAt, AP and GGTP activity). The analysis of clinical and biochemical markers comprised 500 patients (236 males and 264 females) aged 15-35, in whom mononucleosis had been diagnosed according to the commonly approved criteria. The clinical signs and symptoms appearing during the first week of the disease, the results of the laboratory findings and the therapy are described in detail. It has been stressed, that the diagnosis of mononucleosis should be considered mainly in young patients with fever persistent for over one week accompanied by inflammatory changes in the throat and enlargement of the lymph nodes, liver and spleen and other pathologic signs and symptoms. Blood picture and the results of non-specific and specific serologic tests are helpful in the final diagnosis of this interesting disease. PMID- 11921729 TI - [Immunomodulatory treatment of lymphopenia after infectious mononucleosis]. AB - Persistent lymphopenia of various intensity has been noted in 45% of persons who had a history of infectIous mononucleosis. The group consisted od 500 patients, (236 males and 264 females). In all of them the diagnosis of mononucleosis had been based on the characteristic clinical signs and symptoms and changes in the blood picture and confirmed by the non-specific and/or specific serologic tests. In 33 persons (10 males and 18 females) the total number of lymphocytes varied from 884 to 992 per 1 cmm, the distribution being as follows: B lymphocytes--146 to 198, CD4--482 to 645 and CD8--245 to 364 per 1 cmm. 18 persons (18 males and 10 females) were given thymosine (TFX) subcutaneously for 30 days in the dose of 0.02 g daily, the remaining 15 (7 males and 8 females) received 3.0 g of isoprinosine per day for the same period. The total lymphocytes count and that of B, CD4 and CD8 was performed before and after the treatment and then followed once a month until 180 days since the treatment had been started. The TFX therapy was repeated after 8-12 months since the end of the first course in 3 patients and that with isoprinosine in 4. The strategy and the examination pattern were the same as before, control examinations being performed every 6 months and final analysis was done after 5 years of follow-up. Among the 33 patients who received immunomodulating treatment, normalisation of the lymphocytes count appeared in 24 (64%). Among the 12 persons with less pronounced lymphopenia, who did not received any treatment, spontaneous normalisation appeared in 5 (40%). It may be therefore assumed, that in persons with the history of infectious mononucleosis with persistent marked lymphopenia, thymosine and isoprinosine may be used in the therapy as additional immunomodulators. PMID- 11921730 TI - [Recurrent purulent bacterial meningoencephalitis]. AB - During the period of 25 years there were 55 patients treated in our Institute because of recurrent purulent bacterial meningoencephalitis(rpbme). This group consisted of 42 males (76%) and 13 (24%) females, the prevalent number (53%) of patients being under 21 years of age. The diagnosis of rpbme was based on the commonly accepted criteria and confirmed by the laboratory results of CSF examination. The cause of the recurrences was established considering the skull X ray examination, CT and MRI. The evaluation of the clinical status was based on the Glasgow Coma Score (GCS). During the first hospitalisation, severe or critic clinical status was noted in 42 patients (76%) and moderate in 13 (24%). The subsequent recurrences were mostly moderate, rarely severe or mild. The number of recurrences varied from 1 to 9. During the first hospitalisation, the etiologic factor was detected in 39 patients (71%), i.e. Streptococcus pneumoniae in 28 (51%), Neisseria meningitidis in 8 (14%), Pseudomonas aeruginosa and Staphylococcus aureus in 2 and 1 patients respectively. In 37 patients (67%) rpbme developed following cranial trauma, in 18 cases (33%) with single or comminuted fractures of the anterior cranial fossa (in 4 cases accompanied by CSF nasal exsudate). In 4 it followed neurosurgical intervention, in 3 it accompanied recurrent purulent highmorities, in 1 case--after removal of the nasal polyps and subsequent CSF nasal exsudate, and in 1 patient with recurrent mastoiditis. In 6 cases (11%) the cause of the recurrences remained unelucidated. The clinical signs and symptoms, diagnostic difficulties and the causative treatment of rpbme are discussed. In the authors' opinion, surgical treatment of the communication between the CSF and the external environment prevents the recurrences and is the only successful way of treatment. Special attention is drawn to the great diagnostic value of CT and MRI. The use of other modern techniques, e.g. positron emission tomography (PET) is recommended, because it is useful not only in the functional evaluation of the cerebral tissue after the injury, but also in assessing the dynamics of pathologic changes. PMID- 11921731 TI - [Biochemical markers of fibrosis in chronic hepatitis and liver cirrhosis of viral origin]. AB - OBJECTIVE: Usefulness of three selected extracellular matrix components as markers of hepatic fibrosis in patients with chronic viral hepatitis and post inflammatory liver cirrhosis. METHODS: 91 patients with chronic HBV or HCV infection were divided into four groups; chronic hepatitis--group O; cirrhosis classes A, B and C (Child-Pugh classification)--groups A, B and C, respectively. The serum levels of: N-terminal peptide of type III procollagen (PIIIP), laminin (LP1) and hyaluronic acid (HA) in these patients were measured using commercial radioimmunological and radiometric tests. RESULTS: The mean values of the investigated markers increased in patients with cirrhosis according to the stage of the disease (thus group A showed the least level while the highest was exhibited in group C). Between all the groups: O, A, B and C, HA was the marker showing the greatest difference. The only marker with a greater mean level in group O than group A, was PIIIP. Substantially higher levels of the investigated markers were found in the patients with HCV than in those with HBV, but only within group A. CONCLUSIONS: 1) The values of the studied markers increased in various degree with the cirrhosis progress. 2) LP1 and HA measurements have particular value in the clinical estimation of cirrhotic advancement. 3) HA measurement may be helpful in differentiating between cirrhosis and chronic hepatitis. 4) The course of hepatic fibrosis may be influenced by the etiological factor as the values of the studied markers are higher in patients with HCV than in patients with HBV, in the early stage of cirrhosis. PMID- 11921732 TI - [Immunological disorders in patients with chronic viral hepatitis type C]. AB - Various immunological disorders have been observed frequently in patients with chronic viral hepatitis C. Hepatitis C virus has been implicated in the pathogenesis of: essential mixed cryoglobulinemia, cryoglobulinemic vasculitis glomerulonephritis, thrombocytopenia, porphyria cutanea tarda, autoimmune thyroiditis (among others). AIM OF THE STUDY: Was to determine the prevalence and clinical meaning of immunological disorders in HCV infected patients. METHODS: 93 HCV infected patients were studied with regard to the presence of cryoglobulins, autoantibodies, rheumatoid factor (RF) and circulating immunological complexes (CIC). RESULTS: 35 patients out of 93 (38%) had detectable cryoglobulins. Cryoglobulins in 90 cases were of type III and in 3--of type II. CIC /by immunoelectrophoresis of the PEG sediments/ were found in sera of 87 patients (93.5%). 38 persons (41%) had detectable rheumatoid factor. Antinuclear antibodies were found in sera of 15 patients (16%), anti-smooth muscles antibodies--in 21 persons (23%) and anti-LKM1 antibodies--in 5 (5%). Titers of autoantibodies were usually low. The most frequent clinical manifestations were arthralgia and skin changes. CONCLUSION: Immunological disorders /circulating immune complexes (93.5%), autoantibodies (60%), rheumatoid factor (41%), cryoglobulinemia type III (38%)/ are frequently found in HCV infected patients. Age, gender, histological score and clinical evidence of liver cirrhosis do not influence on rate of these abnormalities. Clinical manifestations of immunological disorders are usually mild. PMID- 11921733 TI - [Central nervous system disorders in AIDS patients]. AB - A number of 95 AIDS patients with neurological signs and symptoms hospitalised at the Department of Hepatology and AIDS (in the years 1989-97) were analysed. Sixty patients showed abnormal EEG. Computer assisted tomography (CT) revealed organic changes in them. The remaining 35 patients showed no changes in EEG and CT scan but in 22 of them examination of cerebro-spinal fluid revealed meningoencephalitis of different origin. Moreover, central nervous system autopsy of 55 patients (who had died of AIDS in the years 1986-97) showed macroscopic and/or microscopic changes in 48 out of them (87%). PMID- 11921734 TI - [Hepatitis B virus DNA polymerase]. AB - Our aim was to discuss the practical value of HBV DNA polymerase (DNAp) determination. DNAp activity is a good marker of HBV high replication in patients with chronic hepatitis B (HBeAg+), very useful during monitoring of immunostimulation and/or antiviral treatment. PMID- 11921735 TI - [Usefulness of serological assays in diagnosis of early phase of toxoplasma gondii infection]. AB - OBJECTIVE: The aim of the study was to evaluate value of certain serological assays in differential diagnosis between early and late phase of T. gondii infection. METHODS: 92 patients (58 with nodular toxoplasmosis and 34 asymptomatic women with T. gondii infection diagnosed during or after pregnancy) with different time from onset of infection were tested for T. gondii antibodies with IFA, ISAGA IgM, ELISA IgM, ELISA IgA and IgG avidity assays. Sensitivity and specificity of certain assays was evaluated. RESULTS: IFA had sensitivity 76.19% with specificity 31.67%, ISAGA IgM and ELISA IgM had sensitivity 100% with specificity 16.67% and 23.33% respectively. ELISA IgA had sensitivity 76.19% or 85.71% depending on counting borderline results to positive or negative, with specificity 35% and 25% respectively. IgG avidity had sensitivity 100% with specificity 41.67%. CONCLUSIONS: All evaluated assays have only limited value in differentiation between early and late toxoplasmosis. IgA assay is not superior to IgG and IgM assays. IgG avidity is the most specific and sensitive method of such differentiation. PMID- 11921736 TI - [New trends in ocular toxoplasmosis--the review]. AB - For past ten years some new trends can be observed in the field of ocular toxoplasmosis. In this article on the basis of reviewing scientific articles the author tried to point out important, or new information which may have an impact on clinical researches and clinical practice in the close future. 1. There is evidence that some tissue cysts can be present in untouched retinal tissue, and can produce an inflammatory reaction even many years after primary infection. This concerns both congenital and acquired toxoplasmosis. 2. The coexistence of intraocular inflammatory reaction without focal necrotizing retinochoroiditis in patients with acquired systemic toxoplasmosis was described--but there is no evidence that those inflammations were directly cause by T. gondii parasite themselves. It is too early to include this changes into the classical clinical picture of toxoplasmic retinochoroiditis. 3. For the past ten years there have been published articles showing higher prevalence of acquired ocular toxoplasmosis but this is well documented only in south America. In Europe and USA the frequency of ocular toxoplasmosis cases are low. Despite the different percentage of infected people in these populations, this controversial result can be caused by different distribution of pathogenic and non-pathogenic strains of the parasite as like by differences in human genotypes. 4. There is model of "dynamic equilibrium" existing between host defence and parasite cyst ruptures helpful for understanding reactivation. Level of tissue cyst-specific antigens does not rise in the population of HIV patients or during reactivation of ocular retinochoroiditis--this places the responsibility for reactivation on the side of host immunoresponse. 5. The most specific and reliable laboratory assessment for ocular toxoplasmosis, which is specifically helpful in clinically atypical cases, is combination of IgG, IgA and IgM serology tests made of serum and intraocular fluids. It can proof intraocular production of antibodies. PCR tests have got some value, but they are not as efficient as have been thought previously. 6. There is no efficient treatment diminishing recurrence rate and the time of singular inflammation yet. The combination of pyrimethamine, sulfadiazine and clindamycin, given for 3 to 4 weeks with supplementation of folinic acid for diminishing side effects, can diminish the measure of future scar. Long lasting treatment as it is used in children with congenital disease or in HIV patients can be probably more efficient. Atovaquone is as effective as traditional drugs and gives fewer side effects. PMID- 11921737 TI - [Some aspects of immune response in toxoplasmosis]. AB - OBJECTIVE: The study was performed to determine cell-mediated and humoral responses related to the clinical course of disease in adults with different forms of toxoplasmosis. METHODS: The cellular response was estimated by blastogenic transformation of T-lymphocytes induced by OKT3 mitogenic stimulus, as well as by a specific soluble toxoplasmic antigen. The phenotype analysis of T lymphocytes was performed using monoclonal antibody on FACS flow cytometer. The humoral response was examined by determining the levels of toxoplasma-specific IgA, IgM and IgG antibodies and total serum immunoglobulins. RESULTS: Significant disruption of the cell-mediated response was not observed (T-lymphocyte proliferation test). In the group of patients with lymphadenopathy, an increased proportion of CD8+ lymphocytes was only noted in the first month of the disease, although enlargement of the cervical lymph nodes was observed for up to a year. The individuals in question were found to have IgA antibodies against the p30 antigen of T. gondii, mainly in the period of disease up to 7 months. The proportion of CD4+ lymphocytes was sub-normal in all patients of this group who were studied in the first two months of the disease, and in most of those studied later. Three out of 14 patients with the ocular form of toxoplasmosis only had recent foci of inflammation in the retina, and were also found to have specific IgM. These patients had depressed CD4+ levels, and showed no reaction from CD8+ lymphocytes. CONCLUSIONS: Sub-normal CD4+ population could have been the cause of the weak stimulation of CD8+ cells and an expression of this is the long course of disease. Sub-normal CD4+ cells may be also results the suppressing effect of the parasite. PMID- 11921738 TI - [Hepatitis C--epidemiology, diagnosis and treatment in children]. AB - OBJECTIVE: Epidemiology dates of hepatitis C in children are not well known because of usually asymptomatic course of disease. Vertical transmission has became one of the most common route of hepatitis C virus infection in children. METHODS AND MAIN OBSERVATIONS: Twenty one infants of twenty HCV positive mothers were studied from 1998 to 2000 in the Clinic of Infectious Disease in Childhood of Medical University in Warsaw. Five of the infants were HCV RNA positive. All uninfected children became HCV-antybody negative by 12 months. RESULTS AND CONCLUSIONS: Chronic infection is common in most cases of HCV infection, but the disease progression is slower than in adults. Experience of treatment of chronic hepatitis C in children is limited, with about 40% having sustained response to the interferon therapy. Combinated therapy with interferon and ribavirin may give a better response. PMID- 11921739 TI - [Borrelia burgdorferi infection in children]. AB - OBJECTIVE: The aim of our analysis of causes of borreliosis in children hospitalized or ambulatory treated in the Clinic of Infectious Disease in Childhood of Medical University in Warsaw. METHODS AND MAIN OBSERVATIONS: We observed 113 children from 6 month to 15 years old. Erythema migrans was diagnosed in 97 cases and in 3 cases was multiple erytheme migrans. RESULTS AND CONCLUSIONS: Neuroborreliosis was diagnosed in 11 children. In the treatment neuroboreliosis the ceftriaxon and cristalline penicillin was administrated. PMID- 11921740 TI - [Vertical HIV transmission in Poland from 1989-2000]. AB - OBJECTIVE: To evaluate vertical HIV transmission rate in Poland. METHODS: 132 children born to HIV positive mothers were evaluated and their HIV infection status was established. MAIN OBSERVATIONS AND RESULTS: There are 60 HIV-infected children in Poland. 92% of them have been infected perinataly. From 1989 to 1994 the rate of vertical transmission was 25%. Since September 1994, when prophylactic strategies were started, the rate has decreased to 21.4% (in children with prophylaxis to 1.8%). CONCLUSIONS: Vertical HIV infection prophylactic programs rely on women's knowledge about their HIV infection status and are the only way to diminish pediatric HIV infection worldwide. The authors show difficulty of providing proper medical care of HIV positive women in Poland and underly the need of voluntary HIV testing for all women before pregnancy. PMID- 11921741 TI - [Effects of measles vaccination in HIV infected children]. AB - OBJECTIVE: The aim of the study was to evaluate the effects of measles vaccination in HIV infected children. METHODS: 13 measles seronegative HIV infected children received polyvalent measles-mumps-rubella vaccine. MAIN OBSERVATIONS AND RESULTS: Measles seroconversion occurred in 10/13 patients (in revaccinated children in 7/10), rubella seroconversion in 7 out of 9 patients. None of the 12 children responded mumps component. CONCLUSIONS: Measles vaccination seems to be safe and effective in HIV-infected children without severe immunodeficiency. PMID- 11921743 TI - [Livedo reticularis in patients with systemic lupus erythematosis and its association with anticardiolipin antibodies]. AB - With the purpose of determining whether a relationship exists between Livedo Reticularis (LR) and anticardiolipine antibodies (ACA) in patients with Systemic Lupus Erithemathosus (SLE), a clinical prospective and descriptive study was carried out on 17 female lupus patients with LR, aged 15-46, from the Immunology and Dermatology outpatient clinic of the Enrique Tejera Hospital Compound in Valencia, Venezuela, during 1998. A complete clinical evaluation of each patient was done, from which a diagnosis of LR was made. This was categorized as mild, moderate or severe (Weinstein and col. criteria), depending on the severity of the lesions. Immunoenzymatic techniques were used to determine ACA Ig or IgM. The study followed the Guidelines of Good Practices in Clinical Research, and a signed informed consent was obtained from each patient. The average age of the studied patients was 28.5 +/- 10.9 years; 52.9% of them presented serum levels of ACA IgG above normal. In all cases, levels of ACA IgM were within normal limits (4.41 +/- 2.63 U/mL. Range: 0.51-9.53). All patients with mild LR had normal levels of ACA IgG, and 83.3% and 80% of those in the Moderate and Severe categories, respectively, had high levels of ACA IgG (> or = 10 U/mL), and there was a statistically significant association between each of the three categories and their respective ACA levels (p < 0.05). 82.4% of the patients had severe manifestations of the disease which included: CNS involvement, lower limb vasculitis, renal insult, pericarditis, thrombocytopenia, and recurrent miscarriages. No statistically significant association was found between each of these manifestations and the severity of LR (p > 0.05), probably due to the small number of patients studied in each group; nor between the medium serum levels of ACA IgG and each of the manifestations, except for lower limb vasculitis (ACA IgG'values-presence or absence: Fisher p: < 0.05) However, a statistically significant association (p < 0.05) was observed when the number of severe manifestations of the disease of each patient was related to the levels of ACA IgG, and the severity of LR. PMID- 11921744 TI - [Erythrocytic antigens and their relationship with bipolar disorder. Preliminary study]. AB - There is a close relation between cellular antigens and several diseases. Due to this relation a study was made of the red cell antigen system in 40 patients with Bipolar Disease, who fulfill the diagnostic criteria established by the international classification of Mental Diseases (CIE-10) and 40 control subjects. Known methodology (Orthodiagnostic) was used for the following red cell antigens: ABO, Rh, Hr, MNSs, S, Kidd, Kell, Lewis, Duffy y U. A high prevalence was found in A, RhD, C, Duffy (Fya), Kidd (Jkb), S antigens (p < 0.05), while non significant differences were found in the other antigenic systems studied. It is important to note the close ubication of the locus of the genes, responsible for the antigens with high prevalence, and the susceptibility genes for the bipolar disease. Based in this study it is possible to conclude that persons presenting a high prevalence of the mentioned antigens could be more susceptible to develop the Bipolar Disease. PMID- 11921745 TI - [Vulvovaginitis caused by Candida spp. and Trichomonas vaginalis in sexually active women]. AB - Vulvovaginitis accounts for 20 to 30% of gynecological diseases and it is observed in women from 20 to 30 years of age. It has a higher frequency in women using oral contraceptives and during the third trimester of pregnancy. The aim of this research was to assess the prevalence of Candida spp and Trichomonas vaginalis in patients with the diagnosis of vulvovaginitis from the Gynecology Service in Hospital Universitario "Ruiz y Paez" in Ciudad Bolivar and Hospital "Raul Leoni" in San Felix (Bolivar State, Venezuela). Two hundred women with symptoms of vulvovaginitis were examined, and samples were taken from the uterocervical cul-de-sac. Each patient was asked to fill a questionnaire. The following studies were made in each sample: a) fresh wet mount examination, b) orange acridine and Giemsa stains for Trichomonas and c) culture for the identification of yeasts. The latter were identified by means of the germinal tube assay, resistance to actidione and the presence of clamidospores in rice cream agar and sugar utilization test, using the commercial kit ID32C (BioMerieux). Only in 57 women of 200 patients with vulvovaginitis the causative agent was demonstrated; Candida spp was present in 84.2% (n = 48) and Trichomonas vaginalis in 14% (n = 8). A single case of Zygosaccharomyces spp (1.8%) was detected. The age group mainly affected was that of 25-35 years old, the 38.6% of the studied population. Candida species detected were: C. albicans in 87% of cases (n = 42), C. glabrata, in 10.42% (n = 5), and C. guillermondii, in 2.08 (n = 1). Vaginal flux, vulvar pruritus and leucorrhoea were observed in significant number of patients with vulvovaginal candidiasis. The use of antibiotics was considered predisposing factor for Trichomonas vaginalis infection. The relationship between age and the etiological agent was not statistically significant. Due to the low specificity of clinical manifestations of infections caused by Candida spp and Trichomonas vaginalis, we conclude that performing the etiologic diagnosis of vulvovaginitis is necessary in order to take the appropriate therapeutic and preventive measures, specially in those patients with a recurrent disease. PMID- 11921746 TI - [Tuberculosis in subjects under 15 years of age in the population of Warao in Venezuela]. AB - It is difficult to establish a definitive diagnosis of tuberculosis in rural areas where there is no access to a large hospital. The Warao people of the Delta Amacuro State in Venezuela, have a very high prevalence of adult TB, and we suspected that the Warao children would also have a high prevalence of the disease, almost entirely undiagnosed. We applied a simple methodology to select children suspicious for tuberculosis that is based on a rating system using clinical criteria, reactivity to tuberculin and intradomicilliary contacts. Of the 502 children under the age of 15 that were evaluated with this rating system, 27 were determined to be suspicious of TB and were further evaluated by a chest X ray. Radiologic confirmation of TB was found in 16 (60%) of the 27 suspicious children. Of these 16 patients, 13 (81%) were PPD positive and 3 were PPD negative. Additionally, 7 of the 16 children with pathologic x-ray changes had one or more confirmatory findings: 3 were positive by culture or smear examination and 5 had a positive serologic B diagnostic test. In conclusion this methodology proved to be highly efficient in diagnosing childhood tuberculosis in this population, and should also be useful in other rural populations with a high prevalence of adult TB. PMID- 11921747 TI - Nongranulomatous anterior uveitis associated with alendronate therapy. AB - We describe a case of acute nongranulomatous anterior uveitis associated with alendronate therapy, in an adult woman, without medical history of previous diseases, except for intercurrent problems of osteoporosis. The symptoms disappeared abruptly after anti-inflammatory therapy and discontinuation of alendronate. Side effects associated with ocular inflammation have been recently documented in 3 patients under alendronate therapy. Clinical and laboratory diagnosis of ocular inflammation syndromes are also reviewed. PMID- 11921748 TI - Hunts Point Community Center. The Bronx, New York. PMID- 11921749 TI - Coal Harbour Community Center. Vancouver, British Columbia. PMID- 11921750 TI - Heritage Park Community Center. Chula Vista, California. PMID- 11921751 TI - Building a brand that connects with healthcare consumers. PMID- 11921752 TI - Management challenges under case-mix reimbursement systems. PMID- 11921754 TI - Boost your memory power. PMID- 11921753 TI - Proving the return on your Yellow Pages advertising investment. PMID- 11921755 TI - Debate over a Medicare prescription benefit gains momentum on Capitol Hill. PMID- 11921756 TI - Seven attributes of abuse prevention in long-term care. PMID- 11921757 TI - Quality indicators--blessing or ??? PMID- 11921758 TI - Aroma therapy--help or hinderance. PMID- 11921759 TI - You and your consultant: making the relationship work. PMID- 11921760 TI - Budget bill provides fixes for PPS. PMID- 11921761 TI - Medicare PPS: "fish, or cut bait?". PMID- 11921762 TI - Autogenous tooth transplantation. PMID- 11921763 TI - The combination of a surgical and adhesive restorative approach to treat a deep crown-root fracture: a case report. AB - Most dental trauma requires immediate action to preserve the affected teeth. Furthermore, the trauma has often a great impact on the appearance and well being of the patient. In this case trauma resulted in a very deep complicated crown root fracture of one incisor and a complicated crown fracture in another incisor of a 47-year-old woman. A combination of surgical extrusion and adhesive restoration was chosen to provide good prognosis for the teeth as well as an instant esthetic result that was well accepted by the patient. Eleven months after trauma, the treated teeth exhibited good healing and normal function. PMID- 11921764 TI - Reattachment of an autogenous tooth fragment in a fracture with biologic width violation: a case report. AB - An 11-year-old patient that fractured her maxillary left central incisor is presented. The fracture involved two thirds of the crown, compromising the pulp, and extended subgingivally on the palatal aspect, invading the biologic width. The procedure used to repair the fracture included flap surgery with a slight ostectomy and endodontic treatment. The reattachment of the tooth fragment and the restoration were performed with a bonding system and a resin composite. Examination 6 months after treatment revealed periodontal health, good esthetics, and normal function. PMID- 11921765 TI - Five-year clinical performance of porcelain laminate veneers. AB - OBJECTIVE: The clinical performance of porcelain laminate veneers was evaluated at 5 years. METHOD AND MATERIALS: One hundred eighty-six porcelain laminate veneers were placed in 61 patients, aged 18 to 70 years, by a single operator following the same clinical procedure. At the 5-year recall, esthetics, marginal integrity, marginal discoloration, fracture rate, and patient satisfaction were recorded. RESULTS: At recall 98.4% of the veneers were satisfactory without intervention. The retention rate was excellent, the fracture rate was very low, and the maintenance of esthetics was superior. Patient satisfaction was very high. CONCLUSION: Porcelain laminate veneers offer a reliable and effective procedure for the conservative and esthetic treatment of anterior teeth. PMID- 11921766 TI - Prevalence of interleukin 1 periodontal genotype in a Hispanic dental population. AB - OBJECTIVE: A genetic polymorphism in the interleukin 1 gene has been implicated as a factor in determining the severity of adult periodontitis. Among white Europeans, the prevalence of genotype-positive subjects has been reported to be around 30%. The purpose of this study was to assess the prevalence of genotype positive individuals in a Hispanic population. METHOD AND MATERIALS: Fifty Mexicans were evaluated for their interleukin 1 genotype with a commercially available test. Subjects were divided into groups, as determined by their genotype (positive or negative), and were then analyzed according to age, sex, and smoking habits. RESULTS: Thirteen of 50 subjects were genotype positive, a prevalence of 26%. The most common polymorphisms found in genotype-positive subjects were allele 1.2 for the IL 1A gene and allele 1.2 for the IL 1B gene. When only subjects older than 30 years were evaluated, the prevalence of genotype positive individuals was 31%. CONCLUSION: The prevalence of genotype-positive subjects in a Hispanic population was 26%, similar to the prevalence found among ethnic populations from or descended from Northern, Central, and Southern Europe. PMID- 11921767 TI - Relationship of socioeconomic background to oral hygiene, gingival status, and dental caries in children. AB - OBJECTIVE: The aim of this study was to assess the relationship between socioeconomic status and oral hygiene, gingival condition, and dental caries among 12- to 15-year-old children. METHOD AND MATERIALS: Poor children of low-to moderate socioeconomic status (n = 674) attending 10 public schools were chosen randomly from each of the five geographic areas in Irbid, Jordan. Rich children of high socioeconomic status (n = 347) attending 10 private schools were also included. Schoolchildren were examined for oral hygiene, gingival condition, and dental caries. RESULTS: Significantly higher proportions of children attending public schools had bleeding on brushing and calculus. Mean plaque and gingival scores were higher in public school children than in private school children, but the difference was not statistically significant. The public school children had higher overall scores for decayed, missing, or filled teeth and surfaces as well as higher scores for decayed teeth and surfaces, but there was no statistically significant difference between groups. However, children attending private schools had significantly more missing and filled teeth and surfaces. CONCLUSION: The findings for oral hygiene, gingival status, and dental caries were worse, but not significantly worse, among poor children than they were among rich children. Therefore, dental health education is recommended for both socioeconomic groups. PMID- 11921768 TI - Restoring function and esthetics in a patient with amelogenesis imperfecta: a case report. AB - Amelogenesis imperfecta is a hereditary disorder that affects enamel on primary and permanent teeth. It is a rare dental disease but represents a major restorative challenge for the dentist. A 14-year-old boy presented with sensitive, discolored, and mutilated teeth and a decreased vertical dimension of occlusion. The aim of treatment was to reduce dental sensitivity, to restore esthetics, and to correct the vertical dimension of occlusion. To modify the occlusion, and to protect the dentin from chemical and thermal attacks, nickel chrome onlays were placed on the molars. To improve the esthetics of the incisors and premolars, resin composite restorations were applied. The patient was regularly recalled during the postoperative period. Radiographic and clinical examinations 10 months posttreatment revealed no evidence of disorders associated with the restored teeth or their supporting structures. PMID- 11921770 TI - Dental adhesion: present state of the art and future perspectives. AB - Enamel adhesion by means of acid etching has become an accepted technique in restorative dentistry. Adhesion to dentin, however, is still under investigation. At this time, two distinct adhesive techniques are prominent in the attempt to establish a strong bond to dentin: total-etch and self-etching primers. Total etch bonding systems have been shown to be effective both in vitro and in clinical evaluations. The new total-etch one-bottle bonding systems, although exhibiting the same success on laboratory tests, still have to prove their clinical effectiveness. Self-etching primer systems are undergoing rapid evolution; their results are not yet sufficiently predictable overall, but some systems have achieved positive results in both enamel and dentin bonding. Further studies are necessary to confirm the long-term efficiency of these self-etching primers. PMID- 11921769 TI - Effect of cyclical lateral forces on microleakage in cervical resin composite restorations. AB - OBJECTIVE: The aim of this study was to assess the effect on microleakage of cyclic lateral fatigue forces in different types of cervical resin composite restorations. METHOD AND MATERIALS: V-notch cavities were prepared at the cementoenamel junction in 60 extracted human premolars and restored with a bonding system along with one of the following resin composites: hybrid resin composite, microfilled resin composite, and flowable resin composite (20 each). Ten specimens from each group of 20 were subjected to 8,400 cycles of lateral fatigue forces (44 N) at the occlusal half of the clinical crown in a machine specifically developed for this purpose. The other 10 specimens of each group served as controls, representing the microleakage that occurred without lateral fatigue forces. All specimens were stained with a 50% silver nitrate solution and sectioned sagittally. A scoring system of 0 to 6 was used to record the amount of die penetration along the restoration-tooth interface. RESULTS: There was leakage in both the fatigued specimens and the nonfatigued control specimens. There were significant differences in the mean values of microleakage for the restorative materials studied. In the nonfatigued groups, hybrid resin specimens exhibited significantly more microleakage at the dentin-resin interface than did microfilled or flowable resin specimens. In the fatigued groups, hybrid resin specimens exhibited significantly more leakage at the dentin-resin interface than did microfilled resin specimens. CONCLUSION: The least elastic resin composite tested, the hybrid material, generally showed the most leakage. This suggests that the elasticity of these resin composite restorative materials may be a factor in the amount of microleakage in cervical restorations. PMID- 11921771 TI - Prosthetic rehabilitation of extremely worn dentitions: case reports. AB - The stomatognathic system is a complicated structure, and patients usually adapt to their existing vertical dimension of occlusion. Therefore, prosthetic rehabilitation of decreased vertical dimension of occlusion should be considered only when dictated by esthetic or functional requirements. If the loss of vertical dimension is significant, provisional prostheses should be used before the final prosthesis to prevent tension or pain in the temporomandibular joints and the muscles as the patient adapts to the new dimensions. Acrylic resin provisional prostheses were used for three patients before the final prosthesis was fabricated with the restored vertical dimension of occlusion. The period of use varied, depending on the patient's individual adaptive ability. The important aspect is to find the most comfortable position for the patient. PMID- 11921772 TI - Ectopic impacted mandibular third molar in the subcondylar region associated with a dentigerous cyst: a case report. AB - An impacted third molar in the subcondylar region is extremely rare. This case report describes an impacted mandibular third molar in the subcondylar region associated with a dentigerous cyst. PMID- 11921773 TI - Direct immunofluorescence in oral lichen planus and oral lichenoid reactions. AB - OBJECTIVE: To aid in differentiation of the two lesions, direct immunofluorescence was used to examine the fluorescence patterns in oral lichen planus and oral lichenoid reactions and to compare the degree of intensity of their fluorescence. METHOD AND MATERIALS: Thirty patients participated in this study. Oral mucosal biopsy specimens were obtained from the edge of fresh lesions and were hemisected. One of the sections was placed in 10% neutral buffered formalin for standard light microscopic examination and the other was washed in phosphate-buffered saline and transferred to Michel's transport media for direct immunofluorescence examination. RESULTS: Fibrinogen was deposited in specimens from 14 of 15 patients diagnosed with oral lichen planus; lesions showed an intense positive fluorescence that outlined the basement membrane zone. Eight of 10 sections from oral lichenoid reactions exhibited fibrinogen deposition along the basement membrane zone; the specimens showed a less intense, homogeneous linear fluorescence. Specimens from five control samples showed no fluorescence. CONCLUSION: Fibrinogen was deposited at the basement membrane zone in both oral lichen planus and suspected oral lichenoid reactions, but the fluorescence was less intense in the oral lichenoid lesions. PMID- 11921774 TI - Developmental submandibular salivary gland depression. PMID- 11921775 TI - Advancements in blood-compatible coatings. AB - Heparin-based coatings substantially improve the performance of a variety of blood-contacting medical devices. Recently, it has also been shown that nonheparin-based, synthetic coatings are promising for medical device applications where biological molecules are not desired or appropriate. This article reports on recent work. PMID- 11921776 TI - Interfacial bioengineering to enhance surface biocompatibility. AB - This article reviews recent developments in interfacial bioengineering to improve the biocompatibility of implanted materials. It includes a discussion of biomimetic approaches to bioinert and bioactive surface coatings to improve the in vivo performance of existing medical devices. PMID- 11921777 TI - The technological edge. PMID- 11921778 TI - Implantable microsystems for monitoring and neural rehabilitation, Part II. AB - Miniaturised implantable biomedical microsystems are opening up completely new markets for diagnosis and therapy products. Part II of this article discusses recent work on distributed intelligent implants and biohybrid systems, which combine microsystems with cells and tissues. PMID- 11921779 TI - Direct seal papers for horizontal form-fill-seal machine. AB - Direct-seal papers are designed for minimal fibre lift, where the sealing layer is incorporated into the bottom web. There are applications where they are the most appropriate economical alternative to traditional coated papers for horizontal form-fill-seal packaging. This article explores their benefits and limitations. PMID- 11921780 TI - New US agent requirements. AB - The United States (US) Food and Drug Administration (FDA) has recently published a final rule that will require medical device establishments located outside the US and whose products are imported into the US, to register with FDA and identify a US agent. This article discusses the new establishment registration and US agent requirements. PMID- 11921781 TI - Laser cutting. PMID- 11921782 TI - Innovation and growth in Norway. PMID- 11921783 TI - The enabling technological advantages of chitin. AB - Chitin is a natural biopolymer that, with its derivative chitosan, has been represented as a biomaterial with considerable potential in wide ranging medical applications. This article explains the basis for this claim and describes the nature and properties of these materials. PMID- 11921784 TI - How to simulate shelf life for ageing trials. AB - Medical device manufacturers are compelled to validate the shelf-life claims of their products. If the materials used in the product and packaging can withstand elevated temperatures, accelerated ageing regimes may be used to simulate real time ageing. If elevated temperatures are detrimental to the components involved, then real-time ageing is the only option. All testing should be carried out on sterile product when appropriate. PMID- 11921785 TI - Promoting new ways to provide quality care and service in the emergency department. AB - Today hospital emergency departments are likely to find themselves filled with more patients than ever before. Sometimes, they may have to close their doors temporarily and divert patients to other facilities. However, many hospitals are starting to acknowledge that solutions to ED problems must be initiated systemwide. This means improving staff scheduling, incorporating new information management systems, implementing bedside registration, rethinking triage procedures, communicating better with other hospitals, reviewing patient satisfaction, and improving the way patients are discharged. The results can be improved quality of care--and more satisfied patients. PMID- 11921786 TI - Comment on "Heterodyne lidar returns in the turbulent atmosphere: performance evaluation of simulated systems". AB - The explanation proposed by Belmonte and Rye [Appl. Opt. 39, 2401 (2000)] for the difference between simulation and the zero-order theory for heterodyne lidar returns in a turbulent atmosphere is incorrect. The theoretical expansion the authors considered is not developed under a square-law structure-function approximation (random-wedge atmosphere). Agreement between the simulations and the zero-order term of the theoretical expansion is produced for the limit of statistically independent paths (bistatic operation with large transmitter receiver separation) when the simulations correctly include the large-scale gradients of the turbulent atmosphere. PMID- 11921787 TI - Radiances simulated in the presence of clouds by use of a fast radiative transfer model and a multiple-scattering scheme. AB - A fast-forward radiative transfer (RTF) model is presented that includes cloud radiation interaction for any number of cloud layers. Layer cloud fraction and transmittance are treated separately and combined with that of gaseous transmittances. RTF is tested against a reference procedure that uses line-by line gaseous transmittances and solves the radiative transfer equation by use of the adding-doubling method to handle multiple-scattering conditions properly. The comparison is carried out for channels 8, 12, and 14 of the High Resolution Infrared Radiation Sounder (HIRS/2) and for the geostationary satellite METEOSAT thermal infrared and water vapor channels. Fairly large differences in simulated radiances by the two schemes are found in clear conditions for upper- and mid tropospheric channels; the cause of the differences is discussed. For cloudy situations an improved layer source function is shown to be required when rapid changes in atmospheric transmission are experienced within the model layers. The roles of scattering processes are discussed; results with and without scattering, both obtained by use of a reference code, are compared. Overall, the presented results show that the fast model is capable of reproducing the cloudy results of the much more complex and time-consuming reference scheme. PMID- 11921788 TI - NOAA-NASA Coastal Zone Color Scanner reanalysis effort. AB - Satellite observations of global ocean chlorophyll span more than two decades. However, incompatibilities between processing algorithms prevent us from quantifying natural variability. We applied a comprehensive reanalysis to the Coastal Zone Color Scanner (CZCS) archive, called the National Oceanic and Atmospheric Administration and National Aeronautics and Space Administration (NOAA-NASA) CZCS reanalysis (NCR) effort. NCR consisted of (1) algorithm improvement (AI), where CZCS processing algorithms were improved with modernized atmospheric correction and bio-optical algorithms and (2) blending where in situ data were incorporated into the CZCS AI to minimize residual errors. Global spatial and seasonal patterns of NCR chlorophyll indicated remarkable correspondence with modern sensors, suggesting compatibility. The NCR permits quantitative analyses of interannual and interdecadal trends in global ocean chlorophyll. PMID- 11921789 TI - Retrieval of the ultraviolet aerosol optical depth during a spring campaign in the Bavarian Alps. AB - A measurement campaign was organized in March 1999 in the Bavarian Alps as part of the European project, Characteristics of the UV Radiation Field in the Alps (CUVRA), to analyze the effect of altitude, aerosols, and snow cover on ground level UV spectral irradiance. We present the results of simultaneous measurements of aerosol optical depth (AOD) made at various sites on two cloudless days in March 1999. The two days exhibited different aerosol conditions. Results derived from spectral measurements of UV irradiance are compared with data from filter radiometer measurements made at discrete wavelengths extending from the UV to the near IR. The different methods generated values for the AOD that were in good agreement. This result confirms that one can use either method to retrieve the AOD with an uncertainty of approximately 0.03-0.05. On 18 March, high turbidity was observed at low altitude (400-nm AOD approximately 0.5 at 700 m above sea level), and the AOD decreased regularly with altitude; on 24 March, the turbidity was much less (0.11 at 700 m above sea level). On both days very low AODs (0.05 0.09) were measured at 3000 m above sea level. The spectral dependence of the AOD is often parameterized by the angstrom relationship; the alpha parameter is generally difficult or impossible to retrieve from spectral measurements because of the relatively narrow wavelength range (320-400 nm), and only one of the spectro-radiometers used during the campaign permits this retrieval. In most cases, during this field campaign, alpha was found by filter sunphotometers to be 1.1-1.5. PMID- 11921790 TI - Enhanced autocompensating quantum cryptography system. AB - We have improved the hardware and software of our autocompensating system for quantum key distribution by replacing bulk optical components at the end stations with fiber-optic equivalents and implementing software that synchronizes end station activities, communicates basis choices, corrects errors, and performs privacy amplification over a local area network. The all-fiber-optic arrangement provides stable, efficient, and high-contrast routing of the photons. The low-bit error rate leads to high error-correction efficiency and minimizes data sacrifice during privacy amplification. Characterization measurements made on a number of commercial avalanche photodiodes are presented that highlight the need for improved devices tailored specifically for quantum information applications. A scheme for frequency shifting the photons returning from Alice's station to allow them to be distinguished from backscattered noise photons is also described. PMID- 11921791 TI - Determination of interfacial shear strength between the glass fiber and primary coating in double-coated optical fibers from mechanical strip forces. AB - We propose a method of determining interfacial shear strength between the glass fiber and primary coating in double-coated optical fibers. A commercial stripper is used to mechanically strip the polymeric coatings. It consists of two hinged arms and a pair of blades with a semicircular bore. In the mechanical stripping process the relationship between the strip force and stripper displacement is measured. The interfacial shear strength between the glass fiber and primary coating is then determined from this relationship. The measured interfacial shear strength increases with the increasing stripping rate. It is found that measured results are compatible with theoretical results. PMID- 11921792 TI - Analysis of chirped-sampled and sampled-chirped fiber Bragg gratings. AB - We analyze sampled fiber Bragg gratings that have chirp in the grating period, in the sampling function, or in both. In the last-named case the sampling period can be chirped, the sample length can be chirped, or both. We explain the features in the spectral and group-delay (dispersion) responses for gratings that have a single chirp parameter. We then show how the response of sampled gratings that have multiple chirp parameters can be explained in terms of the single-chirp parameter cases. PMID- 11921793 TI - Multipoint temperature-independent fiber-Bragg-grating strain-sensing system employing an optical-power-detection scheme. AB - A temperature-independent fiber-Bragg-grating strains-sensing system, based on a novel optical-power-detection scheme, is developed and analyzed. In this system a pair of fiber Bragg gratings with reflection spectra either partially or substantially overlapping is placed side by side to form a temperature independent strain-sensor unit. Conventional wavelength-interrogation techniques are not used here, and instead an optical-power-detection scheme is proposed to directly calibrate the measurand, i.e., the strain. Unlike the conventional approach in a multiplexed sensing system, the presented power-detection-based interrogation method does not need the fiber-Bragg-grating sensors to be spectrally separate. The only requirement is that the spectra of the two fiber Bragg gratings of each sensor unit in a multiplexed system be identical or slightly separate (slightly overlapping spectra would also work in the sensing scheme) and the source's optical power be sufficient for sensitive measurement. Based on a three-sensor-unit system, we demonstrate simple strain measurements of high linearity (+/- 0.4%), good sensitivity [2 microstrains (microS)], high thermal stability (+/- 0.8%), and zero cross talk. The effects of light source spectral flatness and fiber bending loss on measurement accuracy are also discussed. PMID- 11921794 TI - Relaxation oscillations in a laser with a Gaussian mirror. AB - We present an analysis of the relaxation oscillations in a laser with a Gaussian mirror by taking into account the three-dimensional spatial field distribution of the laser modes and the spatial hole burning effect. In particular, we discuss the influence of the Gaussian mirror peak reflectivity and a Gaussian parameter on the damping rate and frequency of the relaxation oscillation for two different laser structures, i.e., with a classically unstable resonator and a classically stable resonator. PMID- 11921795 TI - Experimental investigation of saturable gain-guided modes. AB - Hermite-Gaussian modes in geometrically unconfined resonators with a four-level system saturable gain guide inside are experimentally observed and proved. The gain guide was formed with a polished cylindrical surface low-concentration Nd:YAG rod exposed to isotropic pump radiation. The connection of the mode scaling factor with the system parameters is compared with the theory. The abilities of the recently established gain-saturation guiding mechanism to support the generation of beams with wide tops and rapidly decreasing intensity in the wings are also experimentally proved. Application areas for saturable gain guiding are briefly discussed. PMID- 11921796 TI - Design and initial operation of a 367-m2 rectangular ring laser. AB - The design and operation of a proof-of-principle rectangular He-Ne ring laser resonator with a cavity perimeter of 77.0 m and an area of approximately 367 m2 are described. With unevacuated beam lines this device gave an Earth-induced Sagnac frequency of 1513 Hz, with a relative Allan deviation over 1000 s down to 3 parts per million. The Earth's rotation provided a bias that eliminated the lock-in susceptibility. The use of increased pressure in the plasma tube facilitated single-mode operation by increasing the homogeneous pressure broadened linewidth. PMID- 11921797 TI - Laser stability and beam steering in a nonregular polygonal cavity. AB - Two laser stability criteria or lasing conditions for cavity geometry based on the standard ABCD matrix analysis and the Bilger and Stedman analysis [Appl. Opt. 26, 3710 (1987)] are reconciled. Beam steering from mirror misalignment is discussed similarly, generalizing the Bilger and Stedman analysis to nonregular polygons by extending the standard ABCD matrix analysis to 3 x 3 matrices, which facilitates the thorough design of large rectangular ring lasers and is applied to a number of existing or planned ring lasers with perimeters of 77-120 m. PMID- 11921798 TI - Sequence lasing in a gain-switched Yb3+,Er(3+)-doped silica double-clad fiber laser. AB - Experimental results relating to the gain-switched operation of a double-clad Yb3+,Er(3+)-doped silica fiber laser that is pulse pumped with the output from a flash-lamp-pumped Ti:sapphire laser are presented. For all the configurations of the fiber laser that we studied, the 2F5/2-->2F7/2 laser transition of the Yb3+ ion lased prior to laser emission from the 4I13/2-->4I15/2 transition of the Er3+ ion. To the best of our knowledge, this is the first reported operation of sequence lasing in the Yb3+,Er(3+)-codoped system. This succession of laser pulses deduced from the measurements of this investigation is a consequence of both the short intense pump pulse and the short 900-nm wavelength of the pump that does not overlap with any important excited-state absorption transitions. We believe that the predominant interionic interaction during the course of the pump pulse is the double-energy transfer to the Er3+ ion acting twice from the 2F5/2 energy level of the Yb3+ donor ion. A maximum total output of 1.65 mJ is obtained (1.38 mJ from the 2F5/2-->2F7/2 transition of Yb3+ and 0.27 mJ from the 4I13/2- >4I15/2 transition of Er3+) from a nonoptimized configuration of the fiber laser. The wavelength of the output from the fiber laser was measured to vary approximately linearly with fiber length from 1040 to 1046 nm for the Yb(3+) based laser and 1535 to 1541 nm for the Er(3+)-based laser. PMID- 11921799 TI - High-repetition-rate, narrow-band dye lasers with water as a solvent for dyes. AB - The performance of a copper vapor laser-pumped narrow-band dye laser in oscillator-amplifier configuration with water-based binary mixture solvents is described. Although oscillator efficiency in water-surfactant (sodium lauryl sulfate) solvent was comparable with that that employed pure ethanolic solvent, amplifier efficiency was found to be lower. Experiments that were carried out with vertically polarized pump beams and either horizontally or vertically polarized signal beams show that, in case of both the pump and signal having orthogonal polarization (horizontal) and same polarization (vertical), the extraction efficiency for both ethanolic and water-micelle media increased substantially from 15.7% to 18.5% and from 10% to 12.5%, respectively. However, the relative difference remained nearly the same, indicating that a slower orientational diffusion of excited dye molecules in a micellar medium is not responsible for a decrease in amplifier efficiency. Amplifier efficiency comparable with that containing ethanolic dye solutions could be obtained with a binary solvent that comprises a mixture of water and about 30% n-propanol. The performances of two efficient dyes, Rhodamine-6G and Kiton Red S, using water based solvents were studied. PMID- 11921800 TI - Compact 2.5-W 10-kHz Nd:YLF-pumped dye laser. AB - A 10-kHz pulse repetition frequency dye laser, end pumped by a Nd:YLF laser, is reported. This laser was tunable from 590 to 655 nm, and up to 2.55 W of output power was obtained at the 609-nm peak tuning wavelength. By inserting an etalon into the dye laser cavity and frequency doubling using a beta-barium borate crystal, we obtained up to 125 mW of 308-nm single-etalon-mode output, which shows potential for the performance of airborne measurements of tropospheric hydroxyl radical concentrations. PMID- 11921801 TI - Precise wavelength control of a single-frequency pulsed Ho:Tm:YLF laser. AB - We demonstrate wavelength control of a single-frequency diode-pumped Ho:Tm:YLF laser by referencing its wavelength to an absorption line of carbon dioxide. We accomplish this wavelength control by injection seeding with a cw Ho:Tm:YLF laser that can be tuned over or stabilized to carbon dioxide or water vapor lines. We show that the pulsed laser can be scanned precisely over an absorption line of carbon dioxide by scanning the injection seed laser wavelength. We locked the pulsed laser to within 18.5 MHz of the absorption line center by stabilizing the injection seed on the line center. The single-frequency pulsed output, intended for use as a transmitter for differential absorption lidar detection of atmospheric carbon dioxide and water vapor and for coherent detection of wind, is 100 mJ per pulse at a 5-Hz repetition rate. PMID- 11921802 TI - kHz dye laser for use with ultrafast laser systems. AB - We have constructed a cavity-dumped dye laser optimized for use with kHz repetition rate ultrafast lasers for performing experiments on atomic and molecular systems. The dye laser is inexpensive, robust, and requires little pump energy, making it ideal for experiments requiring multiple excitations for state preparation. PMID- 11921803 TI - Red-green-blue laser emissions from dye-doped poly(vinyl alcohol) films. AB - A microscope slide acting as a passive waveguide was coated by three separate poly(vinyl alcohol) films that were doped with Coumarin 460, Disodium Fluorescein, and Rhodamine 640 perchlorate. On collinear pumping by a nitrogen laser, these dyes furnished primary red-green-blue laser emissions that were collected and waveguided by the microscope slide but exited from both ends. Frosting the waveguide exit introduced light scattering at the glass-air interface and spatially overlaid the red-green-blue laser emissions that emerged as a uniform white-light beam. PMID- 11921804 TI - Rh:BaTiO3 thin films with large nonlinear optical properties. AB - We report the fabrication and the nonlinear optical properties of Rh-doped BaTiO3 thin films. The films were deposited on SrTiO3 (100) substrates by pulsed-laser deposition. The deposited Rh:BaTiO3 thin films were single phase and c-axis orientation investigated by x-ray diffraction. The films exhibited large nonlinear optical effects, which were determined using Z-scan technique at a wavelength of 532 nm with a laser duration of 10 ns. The real and imaginary parts of the third-order nonlinear susceptibility chi (3) were 5.71 x 10(-7) esu and 9.59 x 10(-8) esu, respectively. The value of Re chi (3) of Rh:BaTiO3 films is much larger than those of several representative nonlinear optical thin films. The results show that Rh:BaTiO3 thin films have great potential applications for nonlinear optical devices. PMID- 11921805 TI - High-range-resolution velocity-estimation techniques for coherent Doppler lidars with exponentially shaped laser pulses. AB - On the basis of an analysis of the autocovariance of the complex heterodyne signal, some novel algorithms are derived and investigated for recovering the nonuniform Doppler-velocity coherent-lidar profiles within the lidar resolution interval conditioned by the sensing laser-pulse length. The case of exponentially shaped sensing laser pulses is considered. The algorithm performance and efficiency are studied and illustrated by computer simulations (based on the use of pulse models and real laser pulses), taking into account the influence of additive noise and radial-velocity fluctuations. It is shown that, at some reasonable number of signal realizations used and with appropriate data processing to suppress the noise effects, the Doppler-velocity profiles can be determined with a considerably shorter resolution interval in comparison with that (usually accepted as a lower bound) determined by the pulse length. PMID- 11921806 TI - Simulation study for cloud detection with space lidars by use of analog detection photomultiplier tubes. AB - Output signal electrons from photomultiplier tubes (PMTs) have neither a Gaussian nor a Poisson distribution because of changes induced by multiplication when the number of input signal photons and dark electrons is fewer than approximately 100. Therefore the assumption of a Gaussian distribution of signal electrons cannot be used in simulations for space lidar observations with PMTs, for which the number of return signal photons is normally small. A theory is introduced for analog detection with PMTs that have Poisson-distributed secondary-electron emission at each dynode stage. The theory is validated by straightforward numerical simulations. It is shown that the multiplication in PMTs is a multiply stochastic Poisson process and that the distribution of output signal electrons can be interpreted basically as Neyman type A. Analysis by the threshold method of cloud detection with a space lidar shows considerable difference between a Gaussian approximation and the exact distribution. The result indicates that the threshold level must be optimized for the exact distribution. Return signals were simulated for a proposed space lidar, and cloud detection with the threshold method was demonstrated. PMID- 11921807 TI - Assessment of a multibeam Fizeau wedge interferometer for Doppler wind lidar. AB - The Fabry-Perot interferometer is the standard instrument for the direct detection Doppler lidar measurement of atmospheric wind speeds. The multibeam Fizeau wedge has some practical advantages over the Fabry-Perot, such as the linear fringe pattern, and is evaluated for this application. The optimal Fizeau must have a resolving power of 10(6) or more. As the multibeam Fizeau wedge is pushed to such high resolving power, the interference fringes of the device become complicated by asymmetry and secondary maxima. A simple condition for the interferometer plate reflectance, optical gap, and wedge angle reveals whether a set of parameters will yield simple, Airy-like fringes or complex Fizeau fringes. Tilting of the Fizeau wedge improves the fringe shape and permits an extension of the regime of Airy-like fringes to higher resolving power. Sufficient resolving power for the wind lidar application is shown to be possible with a large-gap, low-finesse multibeam Fizeau wedge. Liabilities of the multibeam Fizeau wedge in the wind lidar application include a smaller acceptance solid angle and calibration sensitivity to localized deviations of the plates from the ideal. PMID- 11921808 TI - Bistatic coherent laser radar signal-to-noise ratio. AB - We investigate the signal-to-noise ratio (SNR) for a bistatic coherent laser radar (CLR) system. With a bistatic configuration, the spatial resolution is determined by the overlap of the transmit beam and the virtual backpropagated local oscillator beam. This eliminates the trade-off between range resolution and the bandwidth of the transmitted pulse inherent in monostatic systems. The presented analysis is completely general in that the expressions can be applied to both monostatic and bistatic CLR systems. The heterodyne SNR is computed under the assumption of untruncated Gaussian optics and untruncated Gaussian beam profiles. The analysis also includes the effects of refractive turbulence. The results show that, for maximum SNR, small transmit and local oscillator beam profiles (e-1 intensity radius) are desired. PMID- 11921809 TI - Detection and characterization of biodeteriogens on stone cultural heritage by fluorescence lidar. AB - Biodeteriogens are an important cause of the weathering of a monument, particularly those made of stone, and their detection at an early stage of development helps to protect the monument from deterioration. Frequent mapping of biodeteriogen accumulation is therefore highly necessary. The use of fluorescence lidar for this purpose was introduced in 1995 and has been developed in subsequent years. Three main aspects emerged during this research: the possibility of discriminating between different biodeteriogen strains, the minimum detectable quantity of biodeteriogens, and the control of the efficiency of biocide treatments. We describe the results of a laboratory experiment devoted to clarifying these three aspects of biodeteriogen monitoring by means of fluorescence lidar. PMID- 11921810 TI - Methods for determining regularization for atmospheric retrieval problems. AB - The atmosphere of Earth has already been investigated by several spaceborne instruments, and several further instruments will be launched, e.g., NASA's Earth Observing System Aura platform and the European Space Agency's Environmental Satellite. To stabilize the results in atmospheric retrievals, constraints are used in the iteration process. Therefore hard constraints (discretization of the retrieval grid) and soft constraints (regularization operators) are included in the retrieval. Tikhonov regularization is often used as a soft constraint. In this study, different types of Tikhonov operator were compared, and several new methods were developed to determine the optimal strength of the constraint operationally. The resulting regularization parameters were applied successfully to an ozone retrieval from simulated nadir sounding spectra like those expected to be measured by the Tropospheric Emission Spectrometer, which is part of the Aura platform. Retrievals were characterized by means of estimated error, averaging kernel, vertical resolution, and degrees of freedom. PMID- 11921811 TI - Raman and Rayleigh holographic lidar. AB - We have designed a novel rotational Raman and Rayleigh lidar system that incorporates a simple holographic optical element. The hologram simultaneously disperses and focuses the backscattered signal light so that narrow spectral features can be isolated and detected with high efficiency. By measuring the relative strength of several nitrogen rotational Raman lines, we can obtain an accurate temperature of the atmosphere at a given altitude without the need for external calibration. Simultaneous photon counting of the Rayleigh backscatter signal permits temperature measurements at much higher altitudes. PMID- 11921812 TI - Methodology for determining aerosol optical depth from Brewer 300-320-nm ozone measurements. AB - With a Brewer spectrophotometer, an estimation of total ozone is made from relative measurements of direct-sun ultraviolet radiation at six wavelengths from 300 to 320 nm. During normal operations, one of six neutral-density filters is selected automatically to maintain the detector in its linear response range. On the basis of these standard direct-sun observations, estimates of aerosol optical depth can be derived, provided that a calibration of the relative measurements is available for each neutral-density filter. To obtain the calibration, we implemented a routine to measure direct-sun signals with a fixed neutral-density filter and applied the Langley method to the measured photon counts. Results show that if a sufficiently large number of cloud-free mornings or afternoons is available, a reliable calibration can be achieved even at sea-level sites that are characterized by large aerosol variability. The derived aerosol optical depths appear consistent with those measured independently by a multifilter rotating shadow-band radiometer. Existing relatively long-term series of direct sun ozone measurements by Brewer instruments may be used for retrieval of aerosol optical depth. PMID- 11921813 TI - Detection of ammonia by photoacoustic spectroscopy with semiconductor lasers. AB - Sensitive photoacoustic detection of ammonia with near-infrared diode lasers (1.53 microns) and a novel differential acoustic resonator is described; a sensitivity of 0.2 parts per million volume (signal-to-noise ratio = 1) is attained. To eliminate adsorption-desorption processes of the polar NH3 molecules, a relatively high gas flow of 300 SCCM was used for the ammonia nitrogen mixture. The results are compared with recent ammonia measurements with a NIR diode and absorption spectroscopy used for detection and photoacoustic experiments performed with an infrared quantum-cascade laser. The performance of the much simpler and more compact setup introduced here was comparable with these previous state-of-the-art measurements. PMID- 11921814 TI - Facility profile. Bringing research and care together. PMID- 11921815 TI - The future is now. Hospitals are planning, designing and building with new technologies in mind. PMID- 11921816 TI - Under surveillance. Is it time to reexamine your facility's security model? PMID- 11921817 TI - Eight deadly blunders. Common mistakes in protecting against Legionella. AB - A year has passed since the Joint Commission on Accreditation of Healthcare Organizations (JCAHO), under EC 1.7, began requiring all accredited hospitals to "reduce the potential for organizational-acquired illness ... managing pathogenic biological agents in cooling towers, domestic hot water and other aerosolizing water systems." PMID- 11921818 TI - Training for keeps. Advice on developing an ES training program. PMID- 11921819 TI - Neonatal cleft lip and cleft palate repair. AB - Facial clefting is one of the most common birth defects and having a child born with this defect can be a devastating experience for parents and family members. Until recently, repair of cleft lip and palate deformities was deferred until the infant had reached 10 weeks of age, 10 pounds in weight, and a hemoglobin level of 10 gm per 100 mL. This criteria eliminated neonates 28 days of age or younger from the possibility of early repair. Waiting until these criteria were achieved often caused problems with parent-infant bonding and infant growth and development. Advances in neonatology and pediatric anesthesia now have made it possible to perform cleft surgery during the neonatal period. This article discusses the anatomy and embryology of cleft deformities and how cleft repair surgery now can be performed on the neonate. PMID- 11921820 TI - Medialization thyroplasty for unilateral vocal cord paralysis. AB - Unilateral vocal cord paralysis causes disturbances with vocal function and swallowing, to include aspiration. Thyroplasty type I has become one of the preferred surgical treatments for unilateral vocal cord paralysis. It is tolerated better by the patient, improves predictability of surgical results, and provides good voice results when compared to more traditional procedures, such as teflon injection. Use of a premade thyroplasty implant has made it possible to accomplish permanent medial displacement of a paralyzed vocal cord, avoiding the complications of other surgical options. PMID- 11921821 TI - Morality in perioperative nurses. AB - This study examined nurses' moral motivation, character, and action using a Model of Morality for Perioperative Nurses. Influences on moral actions and selected outcomes for surgical patients and perioperative nurses were examined. Results indicate that motivation and character are related directly to the moral actions of perioperative nurses (R = .13 to .31, P < .001). Fourteen percent of the variance in action was explained by motivation, character, self-perceived level of practice, and ethics education. Results suggest that current models do not describe the moral behavior of perioperative nurses adequately. Future research should examine constructs that explain the moral actions of nurses in the perioperative setting. PMID- 11921822 TI - What is in a name--sedation or anesthesia. AB - The Joint Commission on Accreditation of Healthcare Organizations' sedation standards detail the differences between sedation and anesthesia. These standards, however, note that it is important that those applying sedation be trained to rescue patients who may slip from moderate sedation into deep sedation or from deep sedation into anesthesia. PMID- 11921823 TI - Reprocessing single-use medical devices. AB - Health care providers today often can choose between reprocessed single-use devices (SUDs) or SUDs from original equipment manufacturers. The concern about whether reprocessing is safe and should continue was reflected in the US Food and Drug Administration's draft regulations regarding reprocessing and reuse of SUDs; the Government Accounting Office study on SUDs; legislation introduced at both the federal and state levels; and Congressional hearings by the US House of Representatives and the US Senate. This article offers a review of these activities. PMID- 11921824 TI - Understanding incarcerated populations. AB - Approximately 1.3 to 1.8 million people are incarcerated in the United States at any given time. When providing emergency or elective surgical intervention, perioperative nurses may encounter incarcerated individuals, including many who require treatment for traumatic injury. Nurses, therefore, need to educate themselves about this patient population and understand the specific clinical and educational needs of this group. This article explores key concepts involved in caring for incarcerated individuals, including their characteristics and assumptions others make about them. The article concludes with a discussion of educational and policy implications. PMID- 11921825 TI - Providing perioperative care to patients who are incarcerated. PMID- 11921826 TI - Selecting nurses based on behavioral characteristics. PMID- 11921827 TI - A case study of postoperative delirium. PMID- 11921828 TI - Revising the perioperative nursing data set. PMID- 11921829 TI - Monitoring and measuring errors and adverse events across the surgical continuum. PMID- 11921830 TI - Recommended practices for cleaning and caring for surgical instruments and powered equipment. PMID- 11921831 TI - Recommended practices for managing the patient receiving moderate sedation/analgesia. PMID- 11921832 TI - Comparison of two blood collection techniques for measles antibody detection in children. AB - BACKGROUND & OBJECTIVES: Though several seroprevalence and seroconversion studies have been conducted for measles using the filter paper haemagglutination inhibition (FPHAI), very few studies have compared the conventional serum HAI with the FPHAI. The present study was aimed at the evaluation of whole blood samples on filter paper as an alternative to serum specimens for detection of antibodies to measles virus. METHODS: Serum and whole blood samples soaked on filter paper were collected from 165 randomly selected healthy children in the age groups of 1-2 yr. HAI test was performed on both sets of samples and the results compared. RESULTS: Ninety samples that gave a titre of 8 or more by serum HAI also had titres of > or = 8 by FPHAI showing 100 per cent agreement between the two assays. Seventeen samples that had a serum HAI titre of 2 and 4 were missed by FPHAI due to the starting dilution of 8 of the latter. However, FPHAI gave no false positive results compared with serum HAI. INTERPRETATION & CONCLUSION: Collection of the FP samples by finger prick is more acceptable and requires less expertise than venepuncture. Thus, in spite of the small percentage of missed cases by the filter paper method, this technique of sampling was found to be a convenient and reliable alternative to venepuncture, for detection of measles virus antibody especially in large scale seroepidemiological studies. PMID- 11921833 TI - Seroepidemiology of influenza in Pune, India. AB - BACKGROUND & OBJECTIVES: Influenza viruses cause frequent epidemics and periodic pandemics throughout the world due to antigenic variations. Serological data can be useful to determine the disease burden and population immunity and for predicting the likelihood of occurrence and potential severity of subsequent epidemics. We undertook a serological analysis of antibodies against ten influenza virus strains in Pune, India. METHODS: Haemagglutination inhibition (HI) test was done on 619 sera collected between 1997-99 during an age-stratified serosurvey in Pune, India against 10 strains of influenza virus. Overall prevalence and spectrum of HI antibodies against these strains was determined. RESULTS: Antibodies to at least one influenza virus strain was seen in 62 per cent (116/188) of the sera from individuals in the age group 5-15 yr, 77 per cent (85/111) in sera from 16-25 yr, 78 per cent (93/119) from 26-35 yr, 84 per cent (77/92) from 36-45 yr and 93 per cent (101/109) in sera from individuals aged > 45 yr. The antibody spectrum progressively increased with age. Antibodies to the pandemic strain A(H2N2) were absent in the age groups < 25 yr. INTERPRETATION & CONCLUSION: The results indicate that influenza virus infection occurs in a large proportion of individuals in our community and may be responsible for a considerable amount of morbidity and mortality. The study also demonstrates the absence of antibody to A/Singapore/1/57 (H2N2) strain in younger persons < 25 yr of age. The potential of its reintroduction cannot be ruled out as H2 variants are circulating in wild birds and population immunity in humans is decreasing. PMID- 11921834 TI - Serotype distribution & antimicrobial resistance in Streptococcus pneumoniae causing invasive & other infections in south India. AB - BACKGROUND & OBJECTIVES: Streptococcus pneumoniae continues to be a major cause of morbidity and mortality in developing countries. The emerging resistance to some common antibiotics compounds the problem. There arises a need to monitor the resistance pattern and map serotype distribution in different geographic locations. The present study was undertaken to determine the serotype prevalence and antibiotic susceptibility of clinically significant S. pneumoniae isolated from a tertiary care hospital in south India. METHODS: A total of 150 clinical isolates from invasive and other clinically significant pneumococcal infections were serotyped and screened for susceptibility to commonly used antibiotics by standard and modified laboratory procedures. RESULTS: Majority (59.3%) of the isolates belonged to one or other of the serotypes 1, 6, 19, 5, 23 and 7. Serotype 1 was the commonest isolate from patients of meningitis and empyema followed by pneumonia. Nineteen isolates (12.6%) were nonvaccine type. Eleven (7.3%) isolates were relatively resistant to penicillin (minimum inhibitory concentration was between 0.1 and 1 microgram/ml) and 64 were resistant to one or more antibiotics. Resistance was distributed equally among the predominant serotypes. INTERPRETATION & CONCLUSION: The common serotypes responsible for significant infections were similar to those reported in some other studies from India, with minor variations. Resistance to cotrimoxazole and tetracycline was predominant followed by chloramphenicol. Low level resistance to penicillin was observed but no isolate had absolute resistance. This calls for monitoring of resistance and mapping of serotype distribution from various parts of India. PMID- 11921835 TI - Polymerase chain reaction in the diagnosis of Aspergillus endophthalmitis. AB - BACKGROUND & OBJECTIVES: Aspergillus endophthalmitis is the commonest type of vision threatening fungal endophthalmitis encountered in India. Since conventional methods lack sensitivity, we evaluated polymerase chain reaction (PCR) against the conventional mycological methods in the diagnosis of Aspergillus endophthalmitis. METHODS: Twenty-seven intraocular specimens from 22 patients with suspected fungal endophthalmitis (proven as non-bacterial origin) and 10 patients with non-infective intraocular disorders (controls) were tested. The intraocular specimens from these patients were subjected to the conventional methods, viz., microscopy and culture for growth of fungi, as well as PCR for the detection and differentiation of species of Aspergillus. RESULTS: None of the controls were positive by microscopy, culture or PCR. Among the 27 test samples, 4 were positive by culture for Aspergillus species, these were also positive by PCR. In addition, PCR detected and identified Aspergillus species in 2 culture negative specimens. The average time required for culture and identification of Aspergillus was 10 days, whereas PCR needed only 24 h. INTERPRETATION & CONCLUSION: This study indicates that PCR was not only a more sensitive, but also a rapid diagnostic tool compared to the conventional mycological methods in the diagnosis of Aspergillus endophthalmitis. PMID- 11921837 TI - Prenatal diagnosis of chromosomal abnormalities in women with high risk pregnancies. AB - BACKGROUND & OBJECTIVES: Prenatal diagnosis helps in averting the birth of infants with chromosomal abnormalities. Fluorescence in situ hybridization (FISH) has been introduced as a potentially powerful tool in clinical cytogenetics. Several studies have reported successful prenatal diagnosis of chromosomal abnormalities in high risk pregnancies using FISH, however there are no reported studies from an Indian set up. Prenatal diagnosis for the detection of chromosomal abnormalities was carried out by conventional cytogenetics in the present study in the foetuses of high risk pregnancies in women attending a tertiary care facility in north India. These cases were further analyzed using FISH, to test the efficiency and utility of this technique for prenatal detection of common aneuploidies. METHODS: A total of 82 women with high risk pregnancies (81 singleton, 1 twin) were included in the study. Prenatal diagnosis was performed in these women using conventional chromosomal analysis (CCA) and interphase or metaphase FISH on chorionic villus or amniotic fluid or cord blood samples. RESULTS: Chromosomal analysis was successful in 80 instances and uninformative in 2. Abnormal karyotypes were detected in five (6.09%) of these women. FISH was successful in all the subjects and the results were in conformity to the cytogenetic results. In the 2 women where cytogenetic analysis was uninformative, results were given on the basis of interphase FISH. INTERPRETATION & CONCLUSION: The study clearly demonstrates that prenatal diagnosis is useful in the detection of chromosomal abnormalities in foetuses of women with high risk pregnancies. FISH is a powerful sensitive molecular cytogenetic technique, through which specific chromosomal abnormalities can be diagnosed/identified rapidly and accurately and may be used as an adjunct to conventional cytogenetic analysis. PMID- 11921836 TI - Clinical & genetic analysis of four patients with distal upper limb spinal muscular atrophy. AB - BACKGROUND & OBJECTIVES: Distal upper limb spinal muscular atrophy (SMA) is an uncommon segmental variant of SMA. The condition is usually sporadic, affects males more often than females, and manifests late in the second decade of life, remaining confined to the upper limbs. We examined four patients with this form of SMA in order to determine if they carried homozygous deletion mutations in the survival motor neuron (SMN) or neuronal apoptosis inhibitory protein (NAIP) genes that underlie proximal SMA. METHODS: The four patients with distal upper limb SMA were analysed clinically, electrophysiologically and biochemically. Genomic DNA from each of the patients was analysed by restriction enzyme digestion of polymerase chain reaction (PCR) amplification products, as well single stranded conformation polymorphism (SSCP), to detect deletion events of selected exons of the SMN and NAIP genes. RESULTS: The clinical phenotype of the four patients, together with the biochemical and electrophysiological studies, confirmed a diagnosis of distal upper limb SMA. The molecular studies excluded homozygous deletion mutations in these patients as causative of their phenotype. INTERPRETATION & CONCLUSION: The genetic component underlying distal upper limb SMA appears not to involve mutations that are common in proximal SMA patients. It is possible that genes other than SMN and NAIP may be involved, while somatic mosaicism of SMN gene mutations could be implicated in the segmental nature of distal upper limb SMA. PMID- 11921838 TI - Impacted cuspid treatment with magnets. PMID- 11921839 TI - Hans Peter Bimler at age 85. PMID- 11921840 TI - A very dark Tuesday in December. PMID- 11921841 TI - Orthodontic finishing after TMJ disk manipulation and recapture. AB - Orthodontic treatment has as its goal in most patients to achieve a cosmetic and functional result. There are functional goals for all orthodontic treatment and in some few cases the functional outweigh the cosmetic; an example would be a cleft-palate case. Orthodontic finishing to a specific condylar position is not routinely a goal of orthodontic care. The reason for this is that only patients who have pain, dysfunction and a negative change in quality of life from their temporomandibular apparatus need this tangential type of treatment. Research has shown that most patients suffering from a temporomandibular disorder (TMD) have displacement of the temporomandibular disk(s). Research has shown that when the mandibular condyle is repositioned to the Gelb 4/7 position that the temporomandibular joint disk is recaptured to a normal position between 85% and 96% of the time. The Gelb 4/7 position has been equated to the physiologic position of the mandibular condyle in the glenoid fossa. TMJ condyle repositioning to the physiologic position has been correlated to disk recapture proven by magnetic resonance imaging (MRI). A case is shown in which a displacement without reduction is manipulated into reduction and maintained with orthodontic correction. TMJ disk displacement without reduction is usually preceded by TMJ disk displacement with reduction. Pumping of the upper joint compartment can assist in reducing the TMJ disk displacement without reduction. Magnetic resonance imaging before and after the manipulation and after orthodontic treatment are shown. A detailed method of orthodontic finishing that maintains a specific condylar position and TMJ disk recapture is shown. PMID- 11921842 TI - [Aflatoxin M1 in dairy products: a public health program?]. PMID- 11921843 TI - [Evaluation of health hazards in public dental services: a survey of 14 Italian cities]. PMID- 11921844 TI - [Efficacy and efficiency of the rehabilitation treatment of an amputee patient with vascular disease]. PMID- 11921845 TI - [A model of shared analysis of perceived quality of hospital care]. PMID- 11921846 TI - [Developing an instrument for the assessment of the patient perceived quality of hospital care in Italy]. PMID- 11921847 TI - [Breast feeding. A study on the situation in the area of Pavia]. PMID- 11921848 TI - [Benzene concentration in outdoor air: estimation of leukemia risk for the population of the city of Catania]. PMID- 11921849 TI - [Evaluation of the quality of water of 2 water courses in the Varese Province (Lombardy Region): Ticino River and Strona Stream]. PMID- 11921850 TI - Effect of feeding regime on the metabolism of ochratoxin A during the in vitro incubation in buffered rumen fluid from cows. AB - Pure ochratoxin A (OA) was added to buffered rumen fluid collected from two fistulated cows and incubated under anaerobic conditions. Both animals were fed six diets containing grass, grass silage or hay, and two different amounts of concentrate consisting of barley and soybean meal. Four incubations per animal and diet were carried out at consecutive days. The concentration of OA declined exponentially to a very low or non-detectable level under all conditions examined, with half-lives at 0.51 to 2.76 h. The disappearance of OA was accompanied by the appearance of ochratoxin alpha (O alpha) with an average amount of O alpha formed relative to the disappearance of OA near 100%, independent of diet and animal. Based on four incubations per animal and diet the rate of OA disappearance was affected (P < 0.1) by the origin of rumen fluid from two animals, as well as by the type of basic component and amount of concentrate in the diet, with interactions between these factors. The disappearance of OA mostly was accelerated (P < 0.1) by replacing grass silage or hay by fresh grass and by increasing the content of concentrate from 10 to 50% of dry matter. It is concluded that the capacity of the rumen to detoxify OA is not limited by the yield of Oa from OA but is strongly dependent on animal and diet. PMID- 11921851 TI - Effect of the methodology on peptide amino acid concentrations in blood and plasma of sheep. AB - Different methodologies for the measurement of peptide amino acid (PAA) in blood and plasma were compared in sheep. Preparation of blood and plasma samples consisted of a deproteinization, either chemical with sulfosalicylic acid (0.04 g for 1 ml of sample) or physical by ultrafiltration (10,000-MW cut-off filters), with or without a subsequent ultrafiltration through a 3,000-MW cut-off filter. Peptide concentrations were determined by quantification of amino acid concentrations before and after acid hydrolysis of samples. Free amino acid concentrations were similar by all the method used (about 2.5 and 2.7 mM, for blood and plasma respectively). Peptide concentrations were higher with chemical deproteinization (10.6 and 4.2 mM, for blood and plasma respectively) than with physical deproteinization (5.7 and 3.3 mM, for blood and plasma respectively). When the deproteinized samples were further treated to remove material of molecular weight above than 3 kDa, peptide concentrations were significantly reduced, which indicates inefficiencies in the ability of the deproteinizing procedures in removing all the proteinaceous materials. Concentration of small PAA (< 3 kDa) in blood was about 1.5-fold that in plasma, mainly due to peptide Gly and Glu derived from the hydrolysis of the erythrocyte glutathione. The choice of a methodology for quantifying circulating peptides is discussed. PMID- 11921852 TI - The apparent digestibility of fibre in trotters when dietary soybean oil is substituted for an iso-energetic amount of glucose. AB - An attempt was made to quantify the effect of extra fat intake on fibre utilization in horses. In a 4 x 4 cross-over trial with feeding periods of 24 days each, eight mature trotting horses (age 4 to 12 years, 407 to 531 kg BW) were given four diets. The concentrates were formulated to contain either soybean oil or an iso-energetic amount of glucose or combinations of the two ingredients. The concentrates were fed in combination with the same amount of hay so that the whole diets contained 30, 50, 77 or 108 g EE/kg DM. Apart from the amounts of fat and glucose the four diets were identical. With an increase of 10 g/kg DM of soybean oil the apparent total tract digestibility of crude fibre was reduced with 0.9 percentage units. Extra fat intake also reduced apparent protein and NFE digestibility, but raised apparent fat digestibility. Although the present results may hold specifically for the conditions of this study, it is suggested that the observed interaction between fat content of the diet and macronutrient utilization might have consequences for practical horse feeding in that calculating the energy content of high-fat diets on the basis of feedstuff tables will lead to over- or underestimating the amount of energy provided by the various ingredients of the diets. PMID- 11921853 TI - Effect of short-term underfeeding on weight of splanchnic organs in ewes. AB - We assessed the effect of short-term underfeeding on weight and protein mass of splanchnic tissues in adult ewes submitted to a factorial experimental design. In a pre-experimental period, 18 ewes divided into 2 groups of 9 were fed a second cut of natural grassland hay during 4 weeks at 112 or 38% of their energy maintenance requirements. Three ewes of each group were then fed the same hay during 4 weeks at either low (group L), moderate (group M) or high intake (group H), corresponding to 38, 75 or 112% of their energy maintenance requirements, respectively, then slaughtered. Fresh weight and protein mass of splanchnic organs were measured. No carry-over effect of the pre-experimental level of intake on weight and protein mass was observed for any splanchnic organs. Splanchnic tissues contributed at 10.7 and 8.6% to the decrease in live weight for groups M and L, respectively. The decrease in weight of splanchnic tissues ( 15%) was mainly attributed to reticulorumen (-23%), liver (-21%), and at a lesser extent small intestine (-11%). No large change in the mass ratio between mucosa and muscular-serosa in both dorsal and ventral sacs occurred in the rumen wall. These results are discussed together with previous published results on long-term underfed ewes, and suggest that the decrease in energy expenditure in whole animal induced by underfeeding is mainly related to the decrease in splanchnic weight at short-term, and to the decrease in other tissues at long-term, splanchnic weight being stabilized. PMID- 11921855 TI - Effect of feeding supplemental copper on performance, fatty acid profile and on cholesterol contents and oxidative stability of meat of rabbits. AB - One hundred and four rabbits, five weeks old at the beginning of the experiment, were divided into four groups according to a feed additive treatment. Rabbits of the 1st, 2nd, 3rd and 4th group were fed a basal granulated feed (control), basal feed supplemented with CuSO4.5H2O at 50 mg Cu.kg-1, basal feed supplemented with 150 mg Cu.kg-1, and the latter feed supplemented with 100 mg.kg-1 vitamin E, respectively. The duration of the experiment was 42 days. Addition of Cu at 150 mg.kg-1 increased weight gain non-significantly by 9.1%. This effect was the most pronounced in the first two weeks of fattening. The lowest mortality was observed in rabbits fed the highest amount of additives (7.7% vs. 19.2% in the control). Rabbits were slaughtered at the age of 11 weeks. Neither treatment influenced proportions of saturated, monounsaturated and polyunsaturated fatty acids in lipids extracted from the loin and hindleg muscles. In rabbits fed the highest amount of copper and vitamin E, the cholesterol concentration was significantly decreased by 13.6% and 17.9% in the loin and hindleg meat, respectively. Effects of Cu added at 50 mg.kg-1 were marginal. Copper had no effect on the oxidative stability of meat, measured as thiobarbituric acid-reactive substances in meat stored at 4 degrees C for 0, 3 and 8 days. Vitamin E added in excess of nutritional requirement improved the oxidative stability of meat. In copper-fed rabbits, Cu accumulated in the liver, but not in muscles. Feeding of the basal feed for 7 days to rabbits previously fed copper sulphate decreased the hepatic Cu concentration by 14.0 to 24.4%. PMID- 11921854 TI - Influencing the immune parameters in germ-free piglets by administration of seal oil with increased content of omega-3 PUFA. AB - Oral administration of oil with an increased content of omega-3 polyunsaturated fatty acids to germ-free piglets resulted in a significant increase in the total values of CD4, CD8 lymphocytes, B lymphocytes, and monocytes, in peripheral blood in comparison with the controls. The metabolic activities of phagocytes as well as the polyclonal activation of lymphocytes were not significantly influenced. The level of growth factor was significantly higher, as determined on the basis of somatomedin in the blood serum. Biochemical indices showed a significant increase in the level of eicosapentaenoic and docosahexaenoic acids in blood serum and the decrease in the level of arachidonic acid at the same time. PMID- 11921856 TI - Research note: a method for studying local differences in ruminal fermentation in dairy cattle. AB - A method was developed for studying local differences in ruminal fermentation. The developed sampler consisted of an acrylic glass container (460 cm3) with an aperture for digesta sampling, which could be opened and closed by the scaled "T" rod. The scale was a reference for defined rumen layers: top, middle, 5 to 10 cm and 25 to 35 cm beneath the top of particles mat, respectively, and bottom 5 to 10 cm above the rumen floor. The repeatability of the method was proved in two rumen cannulated cows. Particle/fluid ratio, pH and sample amount were measured 2 to 2 1/2 h after morning feeding in four replicates each day (over 5 days), rumen layer and animal. No significant differences between replicates were observed. The coefficients of variation (CV) of the particle/fluid ratio varied between 8.7% and 13.6%. Top layer had higher CV than middle and bottom layer. CV of pH ranged between 0.59% and 1.27%. The developed method of sampling showed satisfactory repeatability for investigation of digesta properties and fermentation in different rumen layers. PMID- 11921857 TI - An indelible impression.... PMID- 11921858 TI - Tobacco--Oklahoma's #1 preventable cause of death. PMID- 11921859 TI - There's a dark cloud over Oklahoma--and it's our own smoke. AB - The tragic events of September 11, 2001 galvanized our entire nation into action, and appropriately so. Closer to home, in Oklahoma the same number of lives are lost to the adverse health effects of tobacco each year. Fourteen thousand of our young people, under age 18, become addicted to tobacco products each year. One third will die prematurely from tobacco-caused illness. While 48 states have seen their health improve during the decade of the 90s, Oklahoma's health status has actually declined. While not the only malady confronting our society, tobacco takes a disproportionate toll, paid in lost lives and economic impact. Meanwhile, tobacco companies spend more than $100 million annually promoting their product in Oklahoma, 50 percent more than before the state's lawsuit was filed. A two pronged strategy that relies on policy and legislative changes as well as a comprehensive public health program that prevents and treats tobacco addiction is presented. PMID- 11921860 TI - Tobacco wars--the physician's role in public health advocacy. PMID- 11921861 TI - The Oklahoma Alliance on Health or Tobacco. A partnership for a healthier Oklahoma. PMID- 11921862 TI - The Oklahoma Tobacco Settlement Endowment Trust Fund. PMID- 11921863 TI - Second-hand tobacco smoke in Oklahoma: a preventable cause of morbidity and mortality and means of reducing exposure. AB - Evidence has mounted in recent years establishing second-hand tobacco smoke exposure as a cause of morbidity and mortality in nonsmokers. The ratio of deaths is approximately one nonsmoker dying from illness caused by second-hand smoke exposure for every eight smokers who die from diseases caused by tobacco use. This is equivalent to about 750 nonsmoker deaths each year in Oklahoma caused by exposure to second-hand smoke. This article reviews the components of second-hand smoke, its health effects, its prevalence in Oklahoma, and the means of protecting children and nonsmoking adults from exposure. Oklahoma physicians are encouraged to advise their patients about the harmful effects of second-hand smoke and to actively support public policies that decrease exposure to second hand smoke in public places and workplaces. PMID- 11921865 TI - Doctor, are you doing your part to help stamp out tobacco use? PMID- 11921864 TI - Prevalence of current cigarette smoking among American Indians in Oklahoma: a comparison. AB - Tobacco use among American Indians in the US is higher compared to the overall population. Little is known, however, about tobacco use among Native Americans in Oklahoma. The objective of this paper is to report the prevalence of current cigarette smoking among the Native American population in Oklahoma and compare these rates to Oklahoma general adult population rates and United States median rates. The REACH 2010 Native American Behavioral Risk Factor Survey, a random telephone survey, was conducted in 2000 as a part of larger national REACH initiative. It was designed to over-sample Native Americans in Oklahoma, and collect information related to diabetes, cardiovascular disease, overweight and obesity, physical activity, tobacco use, and other behavioral risk factors of interest. The prevalence of current cigarette smoking was significantly higher among Native American adults in Oklahoma (33%) as compared to the Oklahoma general adult population current smoking rate (23%), and U.S. median rate (23%). It was also greater than smoking rates for other racial and/or ethnic groups in Oklahoma. For Native Americans in Oklahoma as well as for Oklahoma and U.S. general populations, highest cigarette smoking rates were among men, younger age groups, and those of lower socioeconomic status, all following the same trend. These findings are urging for more interventions targeting groups with higher smoking rates. PMID- 11921866 TI - Oral cancer examination: perform a death-defying act. PMID- 11921867 TI - Women and children: tobacco or health. PMID- 11921868 TI - Tobacco cessation attitudes and behaviors among Oklahoma high school students. PMID- 11921869 TI - Smoking cessation during pregnancy. AB - Smoking during pregnancy is associated with maternal, fetal, and infant morbidity and mortality. An office-based protocol that systematically identifies pregnant women who smoke and offers treatment has been proven to increase quit rates. For pregnant women who smoke less than 20 cigarettes per day, the provision of a 5-15 minute, five-step counseling session and pregnancy-specific educational materials increases cessation by 30-70%. This bulletin outlines this office-based intervention and addresses treatment issues pertaining to pregnant women who smoke heavily, smoking reduction, pharmacotherapy, healthcare support systems, and coding. PMID- 11921870 TI - Smoking in the elderly--it's never too late to quit. AB - Approximately 3.5 million persons aged 65 years and older smoke cigarettes in the United States--10.9% of this age group in Oklahoma smokes. Up to half of all smokers will experience death or disability related to smoking. Cigarette smoking has been clearly linked to the most common causes of death in the elderly and contributes to the morbidity and disability associated with many chronic illnesses that are common in this age group. The health benefits of smoking cessation in the elderly have been clearly demonstrated. Although there are barriers to smoking cessation that are specific to the older age group, interventions to promote quitting that are successful in younger smokers have been found to be effective in elderly smokers. The extended longevity and quality of life in this country for persons who achieve old age highlights the need for aggressive practices related to smoking cessation in the elderly. PMID- 11921871 TI - Physician knowledge, practice and confidence in treating tobacco use and dependency: Tulsa county physician tobacco survey. PMID- 11921872 TI - Tobacco use prevention and cessation--the focus of the Oklahoma State Medical Association Physicians' Campaign for a Healthier Oklahoma (PCHO). PMID- 11921873 TI - Patient's page. You can quit smoking. PMID- 11921874 TI - "What would a dollar buy?". PMID- 11921875 TI - The extinguisher. PMID- 11921876 TI - The winds of change. PMID- 11921877 TI - [Child and families in the era of reproduction technology--new questions within the scope of child psychiatric treatment]. AB - In the last years the number of children born after assisted human reproduction has increased significantly. These technologies have created new challenges for the families as well as for the professionals involved. This article provides an overview of families after in-vitro-fertilisation, intracytoplasmatic sperm injection and donor insemination. The impact of the specific family composition after DI is described in detail. Two case histories indicate that these families may be confronted with psychiatric and child-development issues. Therefore it is helpful to understand the specific issues of these families. Prospective studies will play an important role in order to understand the quality of parent-child relationship as well as psychological coping strategies, especially in times of crisis. PMID- 11921878 TI - [Invisible fathers: child development and family dynamics after heterologous insemination]. AB - This study presents the long term outcome of the psychotherapy of a girl over seven years, which was born after donor insemination (DI) and aged eight became ill with a compulsive disorder. Aspects of child development, communication within the family and family dynamics related to this specific constellation are pointed out. This case-study presents one of the very few supplements to statistic data elaborated about assitsed-reproduction-families. Implications for individual therapy and therapeutic management of the family are discussed according to development of bonding and identity of the DI-child, father's role, dynamics of the couple and the incognito of the "invisible father". PMID- 11921879 TI - [Subjective quality of life of children and adolescents with psychiatric disorders: development and validation of a new assessment scale]. AB - Within paediatric disciplines and in particular in child and adolescent psychiatry very little work is available regarding the quality of life (QoL) of patients. After using literature search and interviews for identifying the relevant QoL domains, a set of self report questionnaires (for the parents: proxy rating) was developed (C-version: children 10 to 14 years, A-version: adolescents 14 to 18 years, P-version: parental proxy rating about child/adolescent). It was aimed at an almost identical wording within each questionnaire version in regard to the different QoL domains. The questionnaires should not primarily address symptomatology (like e.g. the Child Behaviour Checklist by Achenbach and Edelbrock 1983). After a pilot phase ensuring adequacy and appropriateness of the forms, the questionnaires were field tested in a consecutive series of nearly 700 patients which were referred to the Cologne university department of child and adolescent psychiatry for diagnosis and treatment between 1996 and 2000. The intended structure of the questionnaire system with symptom and functioning scales concerning the domains: "physical performance, fatigue, anxiety, depression, peer group, school, family, beliefs, general QoL" and some further single questions was confirmed by psychometric testing (validity and reliability). Interscale correlations were in general moderate, exhibiting the expected pattern. Analyses of variances using the method of "known groups comparisons" showed e.g. age, sex, and diagnosis related effects hinting towards clinically meaningful differences. The questionnaire forms were well accepted and understood by patients and parents and the wording of the addressed topics was reported to be adequate. Further work will include the psychopathological findings and will focus on specific subgroups of patients (e.g. different diagnoses) as well as on the longitudinal observation of special patient groups, e.g. patients with eating disorders. It can be concluded that the new QoL instruments were successfully employed in a consecutive series of patients and showed very sufficient psychometric properties. PMID- 11921881 TI - Embryonic stem cell research and therapy at European level: is a common legal framework needed? PMID- 11921880 TI - [Discontinuation of treatment in child and adolescent ambulatory psychiatric care in Basel: an evaluation within the scope of quality assurance]. AB - Attrition in child and adolescent psychiatry occurs frequently and has often been investigated. Attempts to establish general criteria that are associated with dropping-out have produced contradictory results. In the present study, factors that influenced attrition at a child psychiatry outpatient clinic in Basel were assessed during two separate periods. Between the two assessments, the results of the first period were presented to the staff and were discussed. At first assessment, attrition was increased in patients who were referred as emergencies or by somatic hospitals. In contrast, attrition was low in patients who were referred involuntarily. At second assessment, there was a significant decrease in dropping-out for the total sample and for emergencies. This result may be explained by a more careful clarification of the setting before first consultation and, as a consequence, by higher congruence between patients' expectations and the clinic's offers. Further studies on the subject of attrition in child and adolescent psychiatry should focus on clearly defined clinical situations and samples and include measures of patients' expectations as well as of the therapeutic relationship. PMID- 11921882 TI - [Moral status of the embryo]. AB - Diagnosis for the purposes of embryo selection prior to implantation, research using embryos and the German embryo Protection Act (1991) constitute the starting point for a discussion of the legal and moral status of embryos. Since the entire genetic programme for a future human being is established at the point of completed union of female egg-cell and male sperm-cell, the embryo conceived in vitro also benefits from the protection of human dignity. It may therefore not be treated exclusively as an object of another's purposes, in other words, it may not be exploited for research or rejected after pre-implantation diagnosis. This conclusion is in accordance with constitutional case-law and international human rights treaties. Research using embryos is permissible in the case of one narrowly circumscribed exception, which will be more closely defined. PMID- 11921883 TI - "Therapeutic" cloning? PMID- 11921884 TI - [Comment on the ruling about the appeal against the Directive on biotechnological inventions]. AB - The author examines the content of the European Court of Justice ruling which dismisses the appeal lodged by the Netherlands against Directive 98/44 concerning the legal protection of biotechnological inventions. The main grounds for the appeal were as follows: inappropriate choice of points of law; breach of the principle of subsidiarity; violation of the principle of legal certainty; breach of International Law obligations; undermining of human dignity; breach of the principle of collegiality. PMID- 11921885 TI - [Awareness about the limit]. AB - Jacquet Testart asked in 1996 "Where are the thinkers, historians, philosophers and pets who are so vital for any reflection on the future of man?" The great Human Genome debate should not be just a technical and learned subject addressed by experts but should embrace society in its entirely. This is a response to Jacquet's call and seeks to equip us for the debate with the weapons of intellectuals and writers. There is still time! PMID- 11921886 TI - Interactive bioethics in a focus group on life and biotechnology in Japan. PMID- 11921887 TI - [Meeting on the biotechnology patent in the reform of the 11/1986 Law]. PMID- 11921889 TI - [Stem cells. The opinion of the European Group on Ethics]. PMID- 11921888 TI - [Conclusions of the seminar on "Biological characteristics, personality and crime: criminological, criminal and procedural features]. PMID- 11921890 TI - The European Group on Ethics in Science and New Technologies to the European Commission. Adoption of an opinion on ethical aspects of human stem cell research and use. PMID- 11921891 TI - Ethical issues in genetic testing for insurance. PMID- 11921892 TI - [Patenting human genetic material: ethical and legal implications]. AB - If we introduce the subject of patents on human genetic material in a Bioethics Conference we must answer two questions. Firstly, whether the debate can be universalized, bearing in mind the national nature of norms governing intellectual property, and, secondly, whether there are links between patent law and ethics. Using the example of the patenting of biological material, we will see how this impacts on society, which, beyond the technical or legal knowledge required, is voicing its concern on the ethical level. PMID- 11921893 TI - Intellectual property and the human genome (Part II). PMID- 11921894 TI - [Liability for prenatal lesions. Rationale, wrongful life and trends (with special attention to Swiss law]. AB - The author provides a detailed consideration of liability for prenatal lesions: causal link, legal fundaments, calculation of compensation, etc. It is argued that the laws of most countries acknowledge that a person suffering post or con conception injury is entitled to seek reparation for bodily or moral injury sustained at the hands of another. PMID- 11921895 TI - [Temporomandibular dysfunction. Introduction]. PMID- 11921896 TI - [Diagnosis and classification of temporomandibular dysfunction by the general dental practitioner]. AB - Because of other diseases mimicking the symptoms of temporomandibular dysfunction (TMD), differential diagnosis is of the utmost importance. Diagnostic subgroups of TMD (osteoarthrosis, anterior disc displacement with/without reduction, TMD myo) can be distinguished by a combination of active movements, passive opening and palpation. Active movement appeared to be the most powerful test for distinguishing the subgroups of TMD, e.g. myogenous, arthrogenous, myogenous and arthrogenous. It is advised to have an open mind in the clinical examination and to be aware of the many mimicking disorders before reaching a diagnosis. An anamnestic questionnaire addressing the pain history, signs and symptoms of TMD, functional impairments, correlates of TMD, psychosocial evaluation and general health status is mandatory. Headache and neck pain often accompany TMD. In appropriate situations it is advised to refer to the proper specialist, before starting treatment for TMD. Orthopedic tests of the cervical spine are of minor importance in discriminating between patients with TMD and patients with cervical spine disorders (CSD). It is therefore advised to evaluate the function of the stomatognathic system in patients with neck complaints to rule out a possible involvement of this system. PMID- 11921897 TI - [Diagnosis and treatment of myofascial pain]. AB - Myofascial pain (MFP) is a regional muscle pain disorder characterized by localized muscle tenderness and pain and is the most common cause of persistent regional pain. MFP is frequently overlooked as a diagnosis because it is often accompanied by signs and symptoms in addition to pain, coincidental pathology conditions, and behavioral and psychosocial problems. Evaluation of myofascial pain includes locating the trigger points and muscles involved as well as recognition of these contributing factors. Management of the syndrome follows with palliative care, splint therapy, muscle exercises, therapy to the trigger points, and behavioral therapy that depends on complexity of the case. The short term goals is to restore the muscle tot normal length, posture, and full joint range of motion with exercises and trigger point therapy. The long term goals include reducing the symptoms and their negative effects while helping the patient return to normal function without need for further health care. The difficulty in managing MFP lies in the critical need to match the level of complexity of the management program with the complexity of the patient. Failure to address the entire problem through a team approach if needed, may lead to failure to resolve the pain and perpetuation of a chronic pain syndrome. PMID- 11921898 TI - [Electromyography. Aid in diagnosis, therapy and therapy evaluation in temporomandibular dysfunction]. AB - Electromyography (EMG) has been applied in studies on patients with temporomandibular dysfunction (TMD), usually with surface electrodes, focusing on rest position of the mandible, hyperactivity and hypo-activity of the muscles, muscle balance, muscle symmetry, reflex activity and parafunctional activity. The contribution of the use of EMG to the diagnostics of individual patients is considered to be marginal, at the most. However, EMG can be valuable in research on TMD if methodological pitfalls are avoided. PMID- 11921899 TI - [Arthroscopy, arthroscopic surgery and arthrocentesis of the temporomandibular joint]. AB - The understanding of the pathogenesis of TMJ disorders is increased by arthroscopic techniques; especially the prominent role of cartilage degeneration has become visible. During arthroscopy a clear view is produced of the different joint surfaces, the anatomic relationship between articular disc and mandibular condyle, and the degree of cartilage degeneration. Arthroscopic surgery results in pain relief, increased joint mobility and function improvement. The arthrocentesis technique is developed based upon the understanding of the underlying therapeutic mechanisms of arthroscopy. The therapeutic effects of arthroscopic procedures and arthrocentesis need to be further evaluated in prospective randomized studies. Future development of arthroscopic techniques will have an increasing impact on the treatment of TMJ disorders. PMID- 11921900 TI - [Neck pain and temporomandibular dysfunction]. AB - Cervical spine disorders (CSD) are common chronic conditions affecting the cervical region and related structures. In this article the coexistence and possible interrelationship between temporomandibular dysfunction (TMD) and CSD is discussed. Our data indicate that a questionnaire is an important aid for distinguishing between CSD patients and subgroups of patients with TMD. Orthopedic tests for the stomatognathic system can be used to discriminate between CSD patients and subgroups of patients with TMD. The orthopedic tests of the cervical spine were shown to be of minor importance in discriminating between patients with TMD and patients with CSD. In more complex cases the cooperation between dentists and 'specialised' physiotherapists can be useful. PMID- 11921901 TI - [Psychosocial factors in patients with temporomandibular dysfunction]. AB - In this article the possible involvement of psychosocial factors in temporomandibular dysfunction (TMD) is discussed, both from a scientific and a clinical point of view. Compared with patients with TMD, controls thought they had personal control over their health and had a more active problem-solving coping style. Patients with a negative treatment outcome were older, reported fewer stressors and more frequently considered health to be determined by external factors than patients who were treated successfully. It was concluded that the assessment of psychosocial variables and in particular health locus of control is important for understanding both subjective treatment need and treatment outcome for TMD. PMID- 11921902 TI - [The use of a questionnaire in TMD]. AB - The use of a questionnaire in a TMD-clinic seems to be a useful aid in diagnosis and treatment of patients with TMD. By using such a questionnaire, it becomes clear which patients have to be advised to attend to psychosocial factors. Besides, the completion of the questionnaire before the first clinical examination often changes the opinion of patients about their complaints. The possible role of psychosocial factors is introduced gradually, and thereby, patients are more receptive to learn adaptive, coping strategies. PMID- 11921903 TI - [Temporomandibular disorders. The role of the otolaryngologist]. AB - Earache, tinnitus, dizziness and hearing loss are frequent complaints of patients with temporomandibular disorder (TMD). Certain patients, therefore, have to be examined by an otolaryngologist. All ear symptoms have to be meticulously described and evaluated by audiometry and vestibular examination. This is obligatory for the evaluation of treatment results and further exploration of the possible relation between these complaints and TMD. PMID- 11921904 TI - [The neurologic differential diagnosis of unilateral headache and facial pain]. AB - The most frequent and relevant neurologic causes of unilateral headache and facial pain are discussed. Symptomatic headache syndromes, caused by an underlying neurologic disease or structural lesion, are distinguished from idiopathic (essential) pain syndromes, which are characterized by a typical clinical presentation and the absence of a known etiology or pathogenesis. PMID- 11921905 TI - [Occlusion and temporomandibular dysfunction]. AB - The role of occlusion in the etiology of temporomandibular disorders is questionable. Therefore, preventive selective grinding is not advised. In the presence of signs and symptoms of overloading of structures of the masticatory system, grinding can be a therapy of choice. This procedure should be preceded by splint therapy in order to test the occlusal changes. PMID- 11921906 TI - [Splint therapy in temporomandibular dysfunction]. AB - In a study of 118 patients with temporomandibular dysfunction treated with occlusal splint therapy, symptoms of pain and limited mobility of the mandible had decreased more distinct than clicking sounds of the temporomandibular joints. Interestingly, an untreated control group showed similar results after two years follow-up. PMID- 11921907 TI - [Temporomandibular dysfunction and chronic rheumatism]. PMID- 11921909 TI - [Intravenous sedation with propofol in dental treatment. Experience in mentally handicapped patients]. AB - Simple dental procedures in mentally handicapped patients can be facilitated by the use of intravenous sedation. This article describes our experiences with eleven such patients undergoing a total of eighteen procedures in 1994-1995. Sedation was administered by a consultant anesthetist using propofol in 'Sterrenberg'--a home for the mentally handicapped in the Netherlands. The article describes the organization of the patients selection and pre- per- and post-sedation care using a management protocol. It is concluded that the technique is effective and safe for simple dental procedures such as examination, removal of tartar, radiography and the treatment of cavities, provided that patients selection and care are meticulous. PMID- 11921910 TI - [Orthodontic treatment and root resorption]. AB - Root resorption after orthodontic treatment occurs frequently. The biologic process of tooth movement and root resorption is described. Furthermore, attention is given to the frequency and severity of root resorption, diagnostics and aspects of the orthodontic treatment which aggravate root resorption. Displacement of teeth with resorptions that already exist or with deviating rootforms is riskfull. PMID- 11921911 TI - [The atrophic maxilla. Rehabilitation by bone reconstruction, implants and permanent or removable dentures]. AB - Maxillary bone reconstruction in combination with placement of endosseous dental implants is a new treatment modality for patients suffering from maxillary atrophy and difficulties with wearing removable prosthesis. Among 88 patients the experiences with and their satisfaction at parts of the treatment were investigated. Patients could express their appreciation in a 1-5 scale (1 = bad, 5 = good). The satisfaction at the total treatment was given the mean figure 4.1 +/- 0.9. Ninety percent of the patients would go through the treatment again when necessary. PMID- 11921912 TI - [Harmony of the facial profile. Part I. Introduction and concept definition]. AB - The terms beauty, attractiveness and harmony can be brought together under the notion of esthetics. Harmony is the only notion to provide us with the possibility of formulating objective and generally applicable criteria for an acceptable facial profile. The prerequisites for these criteria are outlined. PMID- 11921913 TI - [Dental research in the Netherlands: Ph.D theses 1956-1994]. AB - The number of PhD-theses by Dutch scientists shows a continuous increase over the last four decades, resulting in about 20 theses per year during the early nineties. They cover a wide range of subjects in all major disciplines of dental research and nowadays are usually written in English (in 80% of the cases). A substantial proportion (23%) of all dental theses comes from scientists not trained as dentist but in other disciplines like biochemistry and psychology. So far, the drastic decline in the number of Dental Schools in the Netherlands during the eighties has had little influence on the overall outcome of PhD theses. The international impact of Dutch dental research seems to increase over the years as shown by the declining fraction of Dutch dental scientists who are never cited in the international dental literature. PMID- 11921914 TI - [Rational choice? Modifications of the dental health care package]. AB - In a study of the effects of the limitation of the dental health care package, the differences between the public health insured who did apply for supplementary insurance and those who did not were assessed. The motives of both groups were also researched. More then two-thirds of the insured did apply for supplementary dental insurance. Most of this group consist of young, highly educated or high income people who made well considered choices. Of those interviewed 22% did not apply for supplementary dental insurance. On the large, this decision was reached by weighing the costs and benefits. Nearly 10% does not know whether or not they have applied for supplementary dental insurance. High risk groups, such as elderly people, are more often part of this 10%. For some of them the access to dental care seems to be at stake. PMID- 11921916 TI - [Eating disorders]. PMID- 11921915 TI - [An unusual x-ray image in the lower front jaw]. PMID- 11921917 TI - [Cancer, what is it?]. AB - Cancer is characterized by the independent growth of cells based on a series of genetic defects. These include changes in proto-oncogenes and tumor suppressor genes and are based on both pre-existent and acquired alterations in the genome of the cell, caused by exogenous and endogenous factors. The development of cancer is not dependent on a single genetic hit, but requires a series of subsequent alterations. A growing tumour forms its own infrastructure consisting of stroma and blood vessels. Furthermore, the presence of a mononuclear infiltrate indicates an interaction with the immune system. Due to an unbalance between the production and disappearance of cells in a given tissue a growing tumour is born from where cells escape that are capable to penetrate through a basal membrane and that can metastasize to various organs. The increased knowledge regarding the biology of cancer opens new ways for cancer treatment such as gene therapy, immunotherapy and the application of inhibitors of neoangiogenesis. PMID- 11921918 TI - [Epidemiology of cancer in the Netherlands: frequency and prevention]. AB - The Netherlands has a population of approximately 15 million persons. A total number of 55.000 new cancer cases, skin cancer excluded, is registered each year. This incidence figure of approximately 365 per 100.000 population is in accordance with the mean incidence of cancer of all sites of the body in Europe. One out of three men and one out of four women will finally be affected by some form of cancer. Roughly three quarters of the cancer patients are above the age of 60 years. Children form less than one percent of the cancer patient group. Mass screening for cancer seems only feasible in well defined, selected patient groups. PMID- 11921919 TI - [Mouth neoplasms: a review]. AB - An overview is presented of the epidemiology, etiology, clinical and diagnostic aspects, treatment and possible prevention of oral cancer. The dental profession may play a key role in the early detection of cancer and possible precursor lesions such as leukoplakia and erythroplakia. Since the use of tobacco is considered to be the main etiologic factor in oral cancer the dentist can play an active role in its primary prevention by informing his patients about the adverse effect of tobacco use and by encouraging patients to enter tobacco cessation programs. PMID- 11921921 TI - [Radiotherapy: principles and application with special attention to the head and neck region]. AB - Radiotherapy plays an important role in the treatment of cancer and is often applied with curative intent. With the help of modern technical equipment irradiation can be delivered at the tumour localization with maximal sparing of the surrounding normal tissues. Also can differences in the biological effects of irradiation between tumour and normal tissue be utilized to optimize treatment by increasing the chances for cure while limiting side effects. The complex anatomy of the head and neck requires that irradiation is delivered with high precision. Intensive support of the patient before, during, and after treatment is of great importance to control side effects. PMID- 11921920 TI - [Treatment of mouth cancer. General principles as well as surgical aspects]. AB - An overview is presented of the various treatment modalities in head and neck cancer, the emphasis being on oral cancer. Evaluation of the neck with regard to the possible presence of regional lymph node metastases is an integral part of the examination and treatment of patients with oral cancer. A strong plea is made for a multidisciplinary approach in the treatment and rehabilitation of this patient group. PMID- 11921922 TI - [Chemotherapy: principles and application in head and neck tumors]. AB - Chemotherapy is a rational treatment modality when microscopic or macroscopic disease is present. Cytotoxic agents can penetrate in all tissues in the body with exception of brain and gonads. They predominantly affect dividing cells. The role of chemotherapy in squamous cell carcinoma in the head and neck area is limited. Indications where chemotherapy can be considered are recurrence after previous surgery and/or radiotherapy and tumours that are not amenable for curative surgery and/or radiotherapy at first presentation. PMID- 11921923 TI - [Reconstruction possibilities after tumor surgery in the head-neck region]. AB - Surgery and radiotherapy are the most important primary modalities for treatment of head and neck cancer. After excision of a tumour, a considerable defect may remain. Reconstructive surgery may serve several purposes: closure of the defect and restitution of functions such as respiration, swallowing, speech and chewing. Also, an esthetically satisfying appearance is aimed at. The planning of reconstructive surgery starts as soon as a patient presents with a new cancer. The most appropriate procedure is selected by a team of specialists, consisting of head and neck surgeon, a plastic and reconstructive surgeon and a maxillofacial surgeon. For defects of the mouth and oropharynx, a microsurgical free flap is the most versatile and safe choice of reconstruction nowadays. Suitable donor areas are the forearm, iliac crest and abdominal wall and the fibula. The graft, which may consist of skin, fascia, muscle, bone or combinations thereof, is revascularised in the defect area after microsurgical anastomosis of its vascular pedicle. Success percentages are greater than 95% and postoperative morbidity is reduced in contrast to conventional methods. PMID- 11921924 TI - [Oral care in patients receiving radiotherapy and chemotherapy]. AB - Hospitalization of cancer patients is reduced nowadays. The dentist and the oral hygienist in dental practice are involved more and more in minimizing oral complications in cancer patients. Appropriate oral management can enhance the quality of life in patients receiving cancer radiotherapy or chemotherapy. Some of the more significant problems and their prevention or management are discussed in this paper. PMID- 11921925 TI - [Prosthetic reconstruction after oncological surgery in the head and neck region]. AB - The aim of maxillofacial prosthetics is the treatment of patients with head and neck disease in a multidisciplinary way. Intraoral and facial defects are preferably reconstructed by surgical transplantation methods. Patient factors and oncologic factors determine the indication of prosthetic reconstructions. The treatment of patients with an intraoral or facial defect should be based on a close cooperation between maxillofacial prosthetist and MFP technician. The rehabilitation process and the specific role of each are described. PMID- 11921926 TI - [Treatment of mouth cancer at the end of the nineteenth century]. PMID- 11921927 TI - [Theme: local anesthesia in dentistry. Introduction]. PMID- 11921928 TI - [Neuroanatomy and anaesthesiology]. AB - The pathways of nociception, concerning dentogenic pain, are followed from the peripheral nociceptors to the cortex. The branches of the trigeminal nerve supply the semilunar ganglion. From this ganglion the trigeminal nuclei are reached, extending from the bottom of the third ventricle to the upper cervical segments. The thalamus and subsequently the cortex are receiving nociceptive information from trigeminal nuclei. Either ascendent or descendent pathways are involved concerning nociception. PMID- 11921929 TI - [Pharmacology of local anesthetics]. AB - Over the past fifty years, many synthetic local anaesthetics have been developed with fewer side effects, increased specificity of action and a higher margin of safety than cocaine, the first local anaesthetic in clinical medicine. Although the route of administration may affect pharmacokinetics and pharmacodynamics, the present paper focuses on the general pharmacology of these useful compounds. PMID- 11921930 TI - [Internal medicine and local anesthesia in dentistry. A review of possible side effects]. AB - When using local anaesthesia the dentist may be confronted with a variety of side effects, either caused by the anaesthetic solution itself or its additives, including vasoconstrictive drugs such as epinephrine. Early recognition and treatment of these side-effects are essential. Even more important is the possible prevention of such side-effects by a properly taken medical history before using local anaesthesia. PMID- 11921931 TI - [Allergic reactions to local anesthetics]. AB - In this article type I and type IV allergic reactions evoked by local anaesthetics are discussed. Contact dermatitis (type IV) reactions are mainly caused by local anaesthetics of the ester-type like procaine. Both contact dermatitis and anaphylactic reactions (type I) can be caused by anaesthetics of the amide-type (e.g. lidocaine), but these reactions are rare. Intra- and epicutaneous tests can be useful to prove allergy, but sometimes provocation tests are necessary. Many reactions are not caused by an allergic mechanism. PMID- 11921932 TI - [Conduction anesthesia in and around the mouth]. AB - Local infiltration anaesthesia is generally used in dentistry. The so-called nerve block anaesthesia is only routinely used for the mandible. The purpose of this article is to discuss the variety of blocks that can be used by dentists, oral and maxillofacial surgeons and anaesthesiologists. Furthermore, local reactions caused by anaesthetic solutions are described. PMID- 11921933 TI - [Practical aspects of local anesthesia. Infiltration anesthesia]. AB - Local anaesthesia through infiltration is daily practiced in dentistry. Infiltration anaesthesia is effective in gums, mucosa and teeth, in the upper jaw and in the front part of the lower jaw. An amount of 0.3-0.5 ml of anaesthetics including vasoconstrictor is sufficient for 1-2 hours. The method is safe; only few contra-indications are known. Infiltration anaesthesia seldom leads to complications. Intravascular injection cannot entirely be excluded. Therefore slow injection and subsequent observation of the patient's reactions are required. PMID- 11921934 TI - [Local anesthesia in children and mentally handicapped]. AB - Pain belief and pain thresholds vary greatly in children and mentally disabled persons. Whereas adults may tolerate discomfort and remain cooperative, children and mentally disabled persons become frightened when they are hurt and will not cooperate further. Effective pain control is a prerequisite for their successful treatment. There are several ways of producing dental anaesthesia, of which local anaesthesia remains the most widely used technique. The main issue is the preparation of the child and the parents for local anaesthesia. This article offers tools for this preparation, administering and control of local anaesthesia in children and mentally disabled persons. PMID- 11921935 TI - [Local anesthesia in dentistry. Additional techniques]. AB - Patients with insufficient coping abilities can yet be dentally treated by practising several additional techniques. Behaviour management should always be the starting point of all dental treatment strategies. In the end the administration of well selected drugs should lead, via anxiolysis, sedation or general anaesthesia, to a painfree and optimal treatable patient. PMID- 11921936 TI - [Legal aspects of the administration of local anesthesia in dentistry]. AB - In this article the various legal aspects of the administration of local anaesthetics in dentistry are discussed. In general, a capable dentist is not liable for complications resulting from the administration of local anaesthetics, unless the complications could have been foreseen and prevented. PMID- 11921937 TI - [The history of local anesthesia]. PMID- 11921938 TI - [Adjustment of complete dentures. Have oral stereognosis and denture quality a predictable value?]. AB - The denture complaints of 397 complete denture wearing participants were assessed. The quality of the dentures and denture bearing surfaces were professionally scored as well. Combination of these scores gave a specific denture quality, the so-called 'prosthetic condition'. The oral stereognostic capability was recorded with an Oral Stereognosis Test. There was no correlation between denture complaints and the results of the Oral Stereognosis Test, nor between denture complaints and prosthetic condition. PMID- 11921939 TI - [Hematology and dentistry. Part IV. Thrombosis and anticoagulant therapy]. AB - In arterial thrombosis chronic use of acetylsalicylic acid is frequently prescribed. Venous thromboembolism is most often treated with short courses of heparin, followed by oral anticoagulants. All antithrombotic drugs cause hemorrhagic diathesis. Therefore, consultation with the prescribing authority is necessary before proceeding to invasive procedures. PMID- 11921940 TI - [Typing and transmission of periodontal bacteria]. AB - An overview is given of typing methods for bacteria that are associated with periodontitis. Special attention is paid to new molecular biological techniques, based on the characterization of protein and DNA. Bacteria that are associated with periodontitis have a population structure similar to that of opportunistic bacteria. Typing methods are used to study transmission of bacteria. P. gingivalis can be transmitted between spouses, but transmission from parent to child seems to be uncommon. A. actinomycetemcomitans transmission between adults does not occur frequently, while transmission from parent to child can occur in a low frequency. PMID- 11921941 TI - [Supplementary selenium , an antidote against mercury?]. AB - The consequences of both a high and a low intake of selenium are described in this article. Mercury released by dental silver amalgam might affect the protective functions of selenium. However, the literature does not sustain the existence of such an effect. In view of the small margin between safe and toxic doses of selenium and the absence of a scientific consensus as to the possible toxic effects of mercury from amalgam (additional to dietary mercury), it does not seem to be warranted to advise suppletion of selenium. PMID- 11921942 TI - [Dental health of groups at risk in Rotterdam and Amsterdam]. AB - In the Amsterdam and Rotterdam Municipal Health Departments a cohort study at primary schools has been started to evaluate the efficacy of a daily fluoride toothbrushing program and a weekly fluoride mouthrinsing program. Both programs were aimed at groups at risk as far as dental health is concerned. In this article we present the results of the baseline study. At that moment the subjects were six and seven years old. It appears that the average dmfs and the average plaque score were twice as high as those generally found in children of this age group in the Netherlands. If these figures are extrapolated to all primary school children in both cities, it would mean that 40% of the Rotterdam children and 29% of the Amsterdam children are considered to belong to the groups with the highest risk as far as dental health is concerned. These results show that effective preventive measures aimed at the dental health of children are still necessary in the big Dutch cities. PMID- 11921943 TI - [Guidelines for endocarditis prevention. Explanation of the revision of 1996]. PMID- 11921944 TI - [Prevention of bacterial endocarditis. Revision of guidelines for endocarditis prophylaxis 1996]. PMID- 11921946 TI - [Tooth pathology]. PMID- 11921945 TI - [Calculus: prevalence, physiopathology and prevention. Current situation]. PMID- 11921947 TI - [Misleading jaw joint complaints]. PMID- 11921948 TI - [Enamel defects in celiac disease]. PMID- 11921949 TI - [An unusual follicle of the third molar]. PMID- 11921950 TI - [Epilepsy]. PMID- 11921951 TI - [Alveolar ridge augmentation with hydroxylapatite. A clinical evaluation]. AB - In a retrospective study on the effects of an absolute augmentation of the atrophic alveolar ridge with hydroxylapatite, the patient satisfaction rate with the present denture and the loss in height of the augmentation were assessed. Most patients indicated an improved functioning with their dentures after the operation; nevertheless, the patient satisfaction rate was lower than expected. Additionally a concerned loss in height of the augmentation was demonstrated over the evaluation period of 3.5 years. In selected cases, however, the hydroxylapatite augmentation of the atrophic alveolar ridge may still have its value in preprosthetic surgery. PMID- 11921952 TI - [Hematology and dentistry. Part III. Fibrin formation disorder]. AB - After the formation of a platelet-plug, generation of fibrin is necessary for its stabilization. Both congenital and acquired deficiencies of clotting factors occur, leading to retarded formation of fibrin. In congenital disorders, preoperative correction is possible and necessary. In acquired deficiencies, the type and feasibility of correction depends on the cause of the deficiency. PMID- 11921953 TI - [Selenium and mercury. (Un)healthy antagonism?]. AB - The trace element selenium is involved in the protection against damage caused by free radicals. Selenium prevents carcinogenesis and growth of neoplasms. However, the mechanism is insufficiently known. Furthermore, selenium interacts with mercury, thereby preventing toxic reactions to high doses of mercury. The intake of selenium and the desired minimum and maximum concentrations are described. PMID- 11921954 TI - [Orthodontic tooth extraction in a patient with idiopathic thrombocytopenia]. AB - This article describes the removal of a molar in a patient with idiopathic thrombocytopenia by means of orthodontic forces. The indications and advantages as well as the disadvantages of this alternative for surgical extraction are discussed. PMID- 11921955 TI - [How much feedback is provided by the 'feedback post'? Four years of medical information provision to dental practitioners]. AB - This article describes the 'Feedback Post AZIG', an institute to which dentists can turn for advice on medical questions. The frequency as well as the content of these questions, posed between 1991 and 1994, are considered. Dentist clearly have a need for medical feedback and this need increases every year. Given the expected increase of the number of medically compromised patients in the dental practice, the importance of both medical training to dentists and feedback possibilities should not be underestimated. PMID- 11921956 TI - [Practical training in dentistry]. PMID- 11921957 TI - [Treatment of tooth trauma in Dutch general practice. A survey of 52 dentists]. PMID- 11921958 TI - [Pain and fear: a pair apart]. AB - The experience of pain is always affectively negative, and thus often evokes fear. Since the fearful response to pain is not completely determined by the intensity of the pain, dentists should, besides trying to minimize pain, try to minimize this fear. In this article several psychological techniques to achieve that goal are presented and discussed. PMID- 11921959 TI - [Hematology and dentistry. Part II. Abnormalities of the vessel wall and blood platelets]. AB - Haemostasis starts through adhesion of platelets to the subendothelium. Subsequently, ADP and prostaglandins are secreted by the adhered platelets, inducing aggregation of further platelets. The so formed platelet plug stops the bleeding. Congenital disorders of vessel wall and platelets are rare, but acquired abnormalities occur frequently. Usually the consequences of these abnormalities can be handled by local haemostatic action and by the use of antifibrinolytic agents. PMID- 11921960 TI - [Angioedema due to medication treatment]. AB - Patients with hypertension can be treated by inhibition of angiotensin-converting enzyme (ACE). A major side-effect of the use of ACE-inhibitors is the development of a life threatening dyspneu by angioedema of oropharyngeal and laryngeal tissue. This severe complication is caused by interaction of ACE-inhibitors with prostaglandin and bradykinin metabolisms. Minor manifestations of angioedema can be followed by severe reactions. It is wise to stop ACE-inhibitors immediately. PMID- 11921961 TI - [Maxillofacial oncology; quality through cooperation and concentration]. PMID- 11921962 TI - [NVT/NiVVT-Fall Congress 'Pain']. PMID- 11921963 TI - [Infraposition, ankylosis, external root resorption and tooth movement]. PMID- 11921964 TI - [Pain mechanisms become more and more complex]. PMID- 11921965 TI - [Temporomandibular joint sounds: a psychological problem?]. AB - In this article the role of psychological variables in the etiology and presentation of temporomandibular joint sounds is studied. It is concluded that psychological differences observed in the literature are most likely not related to temporomandibular joint sounds. These differences probably are an artefact, caused by using self-report as a substitute for the objective presence of symptoms. A comparison between self-report and objective measurement shows that psychological variables are indeed mainly involved in the reporting of temporomandibular joint sounds. When objective measurements are used, no substantial psychological differences are found. A theory is presented that explains the reporting of symptoms from attention and interpretation. Using this theory the reporting of temporomandibular joint sounds can be predicted quite well. PMID- 11921966 TI - [Degeneration and regeneration. Physiological role in mouth diseases and jaw surgery]. PMID- 11921967 TI - [Toothache?]. PMID- 11921968 TI - [Discoloration of teeth by drugs]. AB - Since 1979 the Dutch Centre for Monitoring of Adverse Reactions to Drugs received 37 reports of tooth discoloration, attributed to the use of drugs. It concerned sixteen males and 21 females with an average age of respectively twelve and 26 years. Most cases were attributed to the use of amoxicillin (n = 16) and doxycycline or minocycline (n = 7). Pseudo-discolorations are chiefly caused by antimicrobial agents, possibly by chromogenic precipitates in the pellicle or by overgrowth with chromogenic micro-organisms. PMID- 11921969 TI - [Mouth and throat cancer; new developments]. PMID- 11921970 TI - [Hematology and dentistry. Part 1. Physiology and mechanisms of hemostasis]. AB - In the dental office, adequate functioning of the haemostatic mechanism is essential in surgical procedures, e.g. tooth extractions. Haemostasis is the result of proper interaction between blood platelets, clotting factors and vessel wall. In the laboratory, intensive testing of the mechanism involved may be performed but the ultimate tests are the patient history and the clinical outcome of invasive procedures. PMID- 11921971 TI - [Skull roof reconstruction in infants]. AB - In this article an introduction into craniofacial surgery in children is given based on the case histories of two patients (cleido-cranial dysplasia and cranio synostosis). PMID- 11921972 TI - [Cost-effectiveness analysis of dental implants]. AB - In this article the cost-effectiveness of dental implants (rootform and transmandibular) is compared with making complete dentures combined with vestibuloplasty and just a set of new dentures. The cost components of labour and technique are analysed at the individual patient level. Effectiveness was measured using a ratio scale. Several assumptions for the long run effectiveness of the treatment options are necessary due to lack of information about this in the literature. Assumptions were made regarding: survival rate, life span of dental implants and complete dentures, costs of aftercare and development of oral health in the long run. The main conclusion is that overdentures on rootform implants are superior to complete dentures combined with preprosthetic surgery or overdentures on a transmandibular implant. The choice between treatment with rootform implants or just a new set of dentures is a more difficult matter. PMID- 11921973 TI - [Antibiotic use in periodontics. Results from a research study]. AB - The sensitivity of the subgingival plaque for amoxicillin, doxycycline, ofloxacin and clindamycin was estimated by agar diffusion disks and Etest in 22 periodontal patients. The sensitivity of the subgingival plaque varied per patient. Resistance of the subgingival plaque for amoxicillin was observed in 50% of the patients and was significantly related to the use of antibiotics in the previous 6 months. The resistance to clindamycin was 68% and to doxycycline 82%. The resistance of the subgingival plaque to these antibiotics could not be related to the previous use. A decision-tree was constructed including a step wise use of a microscopical spirochete test and an antibiotic sensitivity test of the subgingival microflora for the above mentioned antibiotics. The clinical implications of the observed long lasting antibiotic resistance and the practical use of a decision-tree including a spirochete test and antibiotic sensitivity tests need further investigation. PMID- 11921974 TI - [Partial models in fixed prosthodontics]. AB - In general there is consensus regarding the required criteria for crown- and bridge restorations. However, the variety in methods used for their fabrication is large. Textbooks and courses tend to advocate the use of full arch impressions without supplying objective data indicating better clinical behaviour of these restorations compared with the use of quadrant impressions. This article presents advantages, disadvantages, indication and essential steps when using quadrant models. PMID- 11921975 TI - [Hemifacial microsomia. A review]. AB - Hemifacial microsomia is a congenital asymmetrical malformation of the skull and the face. As a congenital malformation of the face hemifacial microsomia is second in prevalence after cleft lip and palate disorders. Combinations and degrees of underdevelopment of craniofacial structures vary greatly. Therapy varies and depends on the different anomalies, but mostly contains orthodontic and surgical treatment to improve symmetry. It is generally accepted that children with asymmetrical facial malformations are best treated in multidisciplinary craniofacial centres. PMID- 11921976 TI - [Harmony of the facial profile. Part II. The analysis]. AB - Rules for harmony of the facial profile reflect the individual perfection of the whole profile. A system of coordinates can be obtained in the same manner in every individual, reflecting the individual form. These coordinates provide valid rules for individual application. PMID- 11921977 TI - [Simple assessment of trigeminal nerve injury]. AB - This paper describes a simple method to assess trigeminal nerve injury. An easy and feasible technique, in particular for the dental surgeon, is scanning the area with altered sensation as indicated by the patient. It is important to refer the eligible patient in time for microsurgical repair of the damaged nerve. PMID- 11921978 TI - [Orthodontics: quality in general practice]. PMID- 11921979 TI - [Water pollution in dental units]. PMID- 11921980 TI - [Dental museum finally has space again]. PMID- 11921981 TI - [An accidental finding: Kahler's disease]. PMID- 11921982 TI - [Hypertension in the dental office]. PMID- 11921983 TI - [Tooth colored dental restorations. Introduction]. PMID- 11921984 TI - [The ideal restorative material]. AB - Dental restorative materials have to fulfill a variety of requirements. Based on practical application, a characterization is presented of the advantages and disadvantages of the different existing materials and which are still in a stage of development. Attention is given to ease of handling and functionality. It is concluded that neither a universally applicable nor an ideal material is available for restorative dentistry. PMID- 11921985 TI - [Damage to the dental pulp by composite bonding systems: leakage or toxicity]. AB - The fourth generation bonding systems demonstrate an excellent adhesion to the dental hard tissues. The marginal seal along the composite-resin restoration does, however, not totally prevent microleakage and, moreover, the bonding system may deteriorate with time. Due to the attention that has been given to possible microleakage, the toxic effects of composite and its components have got less attention in recent years. The present article deals with the question whether the dental pulp is damaged by either microleakage and/or components of the bonding system. PMID- 11921986 TI - [Dental pulp reactions and pulp protection: traditional underlayers versus dentin adhesive lacquer]. AB - The dental pulp possesses an intrinsic healing capacity, by which pulpal reactions remain initially localised; they are often completely reversible. The younger and healthier the pulp is, the greater its healing capacity will be. Only in case of bacterial infection will the sterile pulpal inflammation convert into a local necrosis of the pulpal tissue that gradually will expand apically until the whole pulp is contaminated. The profit-effect of pulpal protection by means of traditional calcium hydroxide medicaments is compared with a more modern adhesive dentin sealing. Potential pulpal response to restorative materials and procedures are reviewed. Finally, clinical guidelines for optimal pulpal protection and dentinal sealing are formulated. PMID- 11921987 TI - [General health damage by restorations of composite]. AB - It seems warranted to state that the fourth generation bonding systems have diminished the probability of bacterial damage to the dental pulp. Dentin and in particular the bonding system may protect the pulp partly at least against toxic side-effects of the numerous components leaching from the composite and against the temperature rise caused by the setting reaction. Generalized toxic side effects caused by the substances released by the restoration and its bonding system are not impossible and are suspected because of in vitro research, but they are assessed to be unlikely in vivo. Generalized and local allergic reactions may appear. The prevalence of allergy to composite may be higher than due to dental silver amalgam, lichenoid lesions excepted. Yet, allergic reactions are not often reported, supposedly because it is either not observed or possibly due to a lack in awareness of such symptoms. PMID- 11921988 TI - [Resin-modified glass-ionomers in dentistry]. AB - The resin-modified glass-ionomers contain the essential components of both a conventional glass-ionomer and a visible light-cured resin. They have several advantages like longer working time, shorter setting time, bonding to tooth structure, fluoride-release, better aesthetics than conventional glass-ionomers and ease in use. As disadvantages can be mentioned the polymerisation shrinkage, trapping of air-bubbles, difficulties in colour choice and contouring. A physical and mechanical characterisation was necessary to define the possibilities and limits of these materials. It can be stated that the strength and aesthetic appearance of resin-modified glass-ionomers are less than that of composites, but better than that of conventional glass-ionomers. Clinically, this is expressed in a rough and dull surface compared to a composite restoration. They are not indicated in stress bearing areas. Consequently, they are mainly indicated in cervical class V-restorations in the postcanine region, and probably restorations in pedodontics, geriatric dentistry and for patients with a high caries activity. PMID- 11921989 TI - [Mechanisms and theory of occlusal wear]. AB - The various mechanisms which cause wear of occlusal surfaces of the teeth, erosion as caused by the food, direct contact wear, and surface fatigue are discussed from their theoretical bases. Special attention is paid to the implications for the oral environment. The ACTA wear machine, designed to simulate these mechanisms, as well as some of its results, are briefly discussed. PMID- 11921990 TI - [Damage to composite restorations]. AB - The expression of damage in dental composite restoratives varies widely, going from some local damage as in attrition facets, to generalized failure or collapes of a restoration. However, crack growth lies on the basis of both aforementioned phenomena. It is therefore of primary importance to gain insight in the way structurally differing materials respond to external stress. It is the aim of the present review to summarize some recent in vitro studies that deal with simulation of wear and fatigue phenomena in dental composite restorative materials. PMID- 11921991 TI - [Restorative treatment of deciduous teeth]. AB - In this article available restoratives for treatment of deciduous teeth are viewed in the light of a theoretical ideal filling material. Especially components and resin composites in combination of a etching primer, approach this ideal in the best way. Clinical studies have to reveal on the long run which material becomes the restorative of choice in the deciduous dentition. For treatment of primary caries in the permanent teeth resin composite offers the best choice. PMID- 11921992 TI - [Preparation forms and filling techniques for posterior composite restorations]. AB - The operator is a very important factor in achieving high quality posterior composite resin restorations. Specific clinical procedures are required to obtain a clinical successful restoration. Tooth saving preparations are made when primary caries is treated. However, tunnel preparations have major disadvantages. The application of a liner or base under a composite restoration is not required anymore since it does not improve the marginal seal when using an total-etch adhesive. The handling and application of the composite should be done with an injection technique and in layers, whereas low viscous composites are giving the best results in preventing voids. PMID- 11921993 TI - [Clinical relevance of fluoride release from restorative materials]. AB - The observed reduced frequency of secondary caries around silicate cement restorations has initiated the development of various fluoride releasing products, to be used as restorative materials, lining cements, sealants and orthodontic cements. The fluoride release is substantial only in the period immediately following usage of the product (e.g. placement of the restoration). An elevation of the fluoride level in plaque or saliva is therefore only found in the immediate vicinity of the fluoride source. Initially, this results in a change of the plaque composition. As a small change in fluoride level affects the caries balance, it is conceivable that fluoride release from dental materials reduces secondary caries, or caries around orthodontic brackets. PMID- 11921994 TI - [Composite inlays]. AB - Composite restorations shrink most during curing. In large cavities more shrinkage may be expected. The composite inlay is especially developed to overcome these disadvantages. After curing of the composite inlay causes secondary polymerization increasing several material properties such as wear resistance and stress relaxation. Clinical results are reviewed and the benefits of the composite inlay technique discussed. PMID- 11921995 TI - [Ceramic inlays and onlays]. AB - Large direct composite restorations can induce shrinkage related postoperative sensitivity. Indirect resin-bonded (tooth colored) restorations may perhaps prevent these complaints. Indirect bonded ceramics are especially attractive because of their biocompatibility and esthetic performance. Several procedures and techniques are currently available for the fabrication of ceramic restorations: firing, casting, heat-pressing and milling. In this article the different systems are described. Advantages, disadvantages and clinical performance of ceramic inlays are compared and discussed. PMID- 11921996 TI - [Composite (etched) bridge]. AB - An adhesive or resin-bonded bridge is a tooth saving construction for the replacement of a lost tooth, especially when the abutment teeth are relatively sound. In this article an overview is presented of the different types of resin bonded bridges, their advantages and disadvantages and their indications. The direct methods are very suited for the immediate replacement of a lost anterior tooth. The all composite adhesive bridge has a survival rate that is surprisingly good. PMID- 11921998 TI - [Badly-controlled diabetes mellitus. A risk factor for dental caries and periodontitis]. PMID- 11921997 TI - [Occlusion-building with tooth-colored restoration materials]. AB - Severe localized or generalized tooth wear will result in loss of vertical dimension. Rehabilitation of the dimensional height is often required. A causal approach is the choice of preference to prevent progression of destructive processes. However, the etiology of pathological wear is not yet clarified, so reversible and non-destructive tooth saving restorative procedures are essential. For adjustment and stabilization of the occlusion of CMD problems the philosophy and treatment approach are similar. Tooth coloured adhesive restorations can be fabricated in different manners, directly and indirectly. In this article treatment objectives for occlusal rehabilitation are described and two cases presented where tooth coloured restorations are used. PMID- 11922000 TI - Storm clouds in Michigan. Blues spars with governor over regulation, reform. PMID- 11921999 TI - But does it work? Measuring effect of staffing ratios could be tough. PMID- 11922001 TI - Talks stall in Florida. Tenet prepares for whistleblower lawsuit. PMID- 11922002 TI - From molehill to mountain. Docs' dispute mushrooms into state probe of hospital. PMID- 11922003 TI - Obesity surgery brings profits. Forest health opens sixth acute-care facility. PMID- 11922005 TI - Lost in the translation. Problem of dirty data about supplies faces cleanup. PMID- 11922004 TI - Nobody said change was easy. North Carolina Blues, state regulators battle over governance, premiums. PMID- 11922006 TI - We're all in the same boat. Don't single out hospitals as cause for skyrocketing healthcare costs. PMID- 11922008 TI - Other investors also on healthcare trail. PMID- 11922007 TI - Make no small plans. Easy-to-track finances of private healthcare services draw attention--and big money--from equity investor firms. PMID- 11922009 TI - Making waves. IRS zeroes in on bond pools, hospital groups' use of tax-exempt financing. AB - The unused debt of an Ohio hospital association has caught the attention of the Internal Revenue Service, which has been on the lookout for bogus pools that take advantage of tax-exempt financing to generate professional fees. Bondholders have been told that the federal agency is examining the loan pool, and similar scenarios may soon occur in other states. PMID- 11922010 TI - Governors seek flexibility, funding for Medicaid. Providers see contributing factors as 'very troubling'. PMID- 11922011 TI - Anesthesia. PMID- 11922012 TI - Office-based general anesthesia in dentistry--yesterday and today. PMID- 11922013 TI - A brief history of anesthesiology in dentistry. PMID- 11922014 TI - Management of the special care patient in a dental surgery center. PMID- 11922015 TI - Accreditation as an alternative to specialty recognition. AB - The Commission on Dental Accreditation does not currently accredit postdoctoral training in general anesthesia. This is related to the traditional linkage of accreditation of postdoctoral training programs to ADA specialty recognition. The aforementioned ADA Guidelines are important because they provide the foundation for the continued use of sedation and general anesthesia within dentistry. The one mechanism that would "ensure safety and effective use," maintain "a strong educational foundation," provide "access to adequate training ... develop programs at the postgraduate level" and support expansion of both basic and clinical research is CDA accreditation of postdoctoral programs. Prior to 1991, hen general anesthesia training was acquired by dentists primarily through accredited medical programs, there was little need for anything more than guidelines to protect the public. With the development of non-accredited dental anesthesiology programs in accredited dental schools (primarily in the late 1980's) and the loss of access to general anesthesia training in accredited medical anesthesiology residency programs in the summer of 1991, primary responsibility for all general anesthesia training of dentists shifted to organized dentistry. However, the ADA House declined to recognize dental anesthesiology as a dental specialty on three separate occasions even though the ADA Council on Dental Education felt all requirements were met and favorably recommended specialty recognition in all three instances. Specialty recognition would have provided the needed accreditation mechanism to validate dentistry's continued use of deep sedation and general anesthesia by those practitioners not classified as oral and maxillofacial surgeons. As an alternative to the specialty recognition process and in view of the need to accredit postdoctoral general anesthesia training for dentists, the American Society of Dentist Anesthesiologists has recently requested the CDA offer accreditation for these dental programs. To address this issue and those of other non-ADA-recognized disciplines, the CDA is currently in the initial stages of developing requirements for accrediting non-ADA recognized postdoctoral training programs. This material will be circulated to communities of interest for their input. Details of the process will be forthcoming from ADA sources in the near future. PMID- 11922017 TI - Oral and maxillofacial pathology case of the month. Eosinophilic granuloma (Langerhans cell disease). PMID- 11922018 TI - The age of enlightenment. PMID- 11922019 TI - Proceedings of the forty-fourth annual coccidioidomycosis study group meeting. Berkeley, California, USA, April 1, 2000. PMID- 11922020 TI - Serum zinc and copper concentrations in Nigerian women with normal pregnancy. AB - The impact of normal pregnancy on serum zinc and copper concentrations and on copper/zinc ratio has been determined in a cross-sectional study of eighty-four healthy pregnant Nigerian women. Fifty-seven healthy non-pregnant women served as controls. Serum zinc and copper levels were determined by atomic absorption spectroscopy. Mean serum copper concentration is significantly higher (P < 0.001) in the pregnant group than in controls [19.74(4.65) and 11.87(2.61) micromol/L respectively], while mean zinc level is significantly lower (P< 0.001) than in controls [4.40(1.34) and 5.54(1.04) micromol/L respectively. Copper/Zinc ratio is also significantly higher than (P<0.001) in the test subject [4.87(1.77) and 2.23(0.75) micromol/L respectively]. One-way analysis of variance shows that the mean serum zinc and copper concentrations and copper/zinc ratio differ significantly (P < 0.01, P < 0.05, and P < 0.05 respectively) between any two of the trimester groups. Furthermore, there is a significant positive correlation between gestational age and copper/zinc ratio (r- 0.201; P < 0.05), and a significant negative correlation between gestational age and serum zinc concentration (r = -0.274; P < 0.01). In normal pregnancy, a negative and positive linear relationship exists between gestational age and serum zinc level and copper/zinc ratio respectively. Serum copper does not show any significant trend throughout gestation, though pregnant subjects have higher mean serum copper level than controls. PMID- 11922021 TI - Direct toxic effects of clinical doses of chloroquine on transferrin secretion in immature rat sertoli cells in vitro. AB - We have examined the effects of increasing doses of chloroquine (CQ), on transferrin secretion in primary cultures of immature rat Sertoli cells (SC) grown on a reconstituted basement membrane (Matrigel) in bicameral chambers. SC cells were seeded in serum-free defined medium at a density of 3 x 10(6) cells/0.64cm2/well on Matrigel covered Millicell-HA filters. CQ at concentrations ranging from 0.04-1.0 microM was added to the basal compartment of the bicameral system from day 7 of the culture. The formation of the tight junction was monitored by the measurement of the transepithelial resistance (TER) at 24 hr intervals using an impedance meter TER in untreated controls was 50 Ohms/cm2 on day 1, and increased progressively to 80 Ohms/cm2 by day 7 and plateaued until day 12. On the seventh day of culture, CQ was introduced into the basal chamber During the 4 days of the experiment, the secretion of transferrin decreased with time. Maximal transferrin secretion by SC was detected during the initial 2 day collection period. During the subsequent collection period, CQ (1 microM) decreased significantly transferrin secretion by SC, while 0.04 microM CQ did not affect transferrin secretion. The polarized secretion of transferrin in response to CQ was also studied. During both collection periods there was no significant difference between controls and 0.04 microM CQ cultures in the ratio of apical to basal transferrin secretion. In the 1 microM culture medium, CQ diminished significantly the ratio of apical to basal transferrin secretion. These observations demonstrate the heterogenous effects of lower doses of CQ on immature rat SC in cultures. PMID- 11922022 TI - Otorhinolaryngological manifestations of HIV/AIDS in Lagos. AB - A prospective study was carried out at the Lagos University Teaching Hospital in Lagos. The aims were to determine the prevalence of otorhinolaryngological disease among patients who were HIV+ and or had clinical AIDS and to correlate the presence of ORL disease with the clinical state of HIV infection. The study was done between October 1998 and September 1999. A total of ninety-eight patients were studied. The age range of the patients was between 15 and 69 years with 83% of them being in the age group of 20-49 years. The results showed that while only 17% of the patients were referred because of ORL diseases, 80% of them actually had O.R.L./head and neck conditions. This difference of proportion is highly significant P = 0.000037. 160.20% of the patients had oral/pharyngeal lesions and 24.5% had identifiable otological disease. Hearing impairment was noted in 30.6% of the patients on pure tone audiometric assessment of which 26.5% were sensorineural. The correlation between O.R.L./head and neck manifestation and the CDC classification of HIV/AIDS infection did not reveal a definite pattern. PMID- 11922023 TI - Management of cancer pain--a survey of current practice in West Africa. AB - A comprehensive management plan for cancer pain is yet to be formulated in the West African sub region despite the priority that the World Health Organisation (WHO) has devoted to the problem over the past decade. As a prerequisite to our cancer pain management curriculum development a structured questionnaire on cancer pain management practice was administered to 80 Fellows attending the Annual Scientific Conference of the West African College of Surgeons in 1996. They were asked how often they treated cancer patients, the causes of cancer pain, their methods of assessment and therapeutic measures and complications of management techniques. Forty-four fellows from Nigeria, Ghana, Liberia and Sierra Leone, all consultants and trainers, responded to the questionnaire. About 80% of them treated cancer patients all the time. Specific anti-cancer therapy such as surgery, chemotherapy and hormone therapy were available in the four countries but radiotherapy was only available in two centers in Nigeria. The respondents estimated that 70-90% of their patients had severe pain at presentation. Pain was thought by 52% of respondents to be due to cancer and its treatment while 47% thought it was due to cancer and the fear of dying. Pain assessment was mostly by the verbal rating scale, only 20% included psychological measurement in their schedules. Oral preparations of strong opioids were not available in most countries andfor severe pain, the parenteral route was employed. Only 18% knew about the 'by the clock' dosing schedule. The study revealed that the standard of practice of the respondents falls below accepted practice. There is thus an urgent need for the formal education of personnel involved in the care of these patients. PMID- 11922024 TI - Medical admission into the intensive care unit (ICU) of the Lagos University Teaching Hospital. AB - The paper presents patterns of medical admissions into the intensive care unit of the Lagos University Teaching Hospital (LUTH) over an eight-year period from September 1990 to August 1998. Medical admission constituted 15% out of which 1% received surgery for medical complications. Patients with neurological diseases recorded the highest number of admission most of them being for tetanus. The commonest indication for admission was for respiratory insufficiency (33%). All the patients admitted for sub arachnoid haemorrhage, fulminant hepatitis, meningitis and motor neurone disease died. The least mortality was found amongst patients admitted for tetanus who constituted about 44% of the total number of medical admissions. The overall mortality rate was 69% and it was observed that the ages of the patients did not appear to affect the outcome except in patients who were admitted for myocardial infarction and cardiogenic shock. This study emphasizes the need to evolve a system of health evaluation of predicting the survival index of individual patients. There is an urgent need for proper training, motivation of staff and maintenance of equipment used in the ICU. PMID- 11922025 TI - Prevalence of trichomonas vaginalis in patients with vaginal discharge in Lagos, Nigeria. AB - Trichomonas vaginalis is the most prevalent, non-viral sexually transmitted infection worldwide and probably the most commonly treatable in Africa and worldwide. The purpose of this study was to determine the prevalence of T. vaginalis among women with vaginal discharge attending a gynaecologic clinic in Lagos and to identify the common characteristics of the vaginal discharge in T Vaginalis. High vaginal swabs were taken from consenting adults with vaginal discharge. The quantity, colour and odour of the discharge were noted. Amine (10% KOH) test and pH were determined. Wet-mount microscopy Giemsa staining and culture in Oxoid (R) Trichomonas Medium were performed on each swab. A total of 200 patients were examined. One hundred and forty-nine (74.5%) had T. vaginalis. There was no statistically significant association between age, marital status, parity, number of sexual partners and prevalence of T vaginalis. The colour of the discharge was white in 104 (69.8%), yellow in 30 (20.1), clear in 15 (10.1%). None was frothy or greenish. The discharge was heavy in 50 (33.6%) and malodorous in 51 (34.2%). The pH range was 4-7 and 42 (28.2%) normal pH of4. In 47 (31.5%) the amine test was negative. The prevalence of T vaginalis among women with vaginal discharge is high. Women complaining of vaginal discharge should be thoroughly screened for T. vaginalis using all available methods. PMID- 11922026 TI - Microalbuminuria and associated factors in newly diagnosed diabetics. AB - Diabetic nephropathy is a common cause of end-stage renal disease. Microalbuminuria, a predictor of diabetic nephropathy, was screened for in newly diagnosed diabetics with the objectives of finding its prevalence and factors associated with its development. The Micral-Test strip was used to screen for microalbuminuria in 66 newly diagnosed diabetics (21 Type 1, 45 Type 2) and 36 non-diabetic controls. Microalbuminuria was present in 50% of diabetics and 8% of controls. Compared with the control group, diabetics had a higher prevalence of smoking, significantly higher mean systolic and diastolic blood pressure and mean arterial pressure. Retinopathy was present in 23% of diabetics and was more frequently encountered in females. From multiple regression analysis smoking, systolic blood pressure, mean arterial pressure and retinopathy were significantly associated with microalbuminuria. In conclusion, microalbuminuria occurs commonly in newly diagnosed diabetics and its development is associated with elevated levels of systolic blood pressure, mean arterial pressure, retinopathy and smoking. Screening for this associated factors of diabetic nephropathy is recommended as a routine in all newly diagnosed diabetics. PMID- 11922027 TI - Prevalence of oral habits in 563 Nigerian preschool children age 3-5 years. AB - One of the contributory factors in the establishment of occlusion is the child's oral habits. A limited amount of information is available on oral habits of pre school children especially on Nigerian pre-school children. With an increasing interest in the early recognition of mal-occlusion and a corresponding emphasis on preventive procedures, more information on pre-school children may prove to be useful. This study revealed the prevalence of oral habits among 563 Nigerian pre school children aged 3-5 years at 13.14%: 6.74% for males, 6.4%for females. The acquired data were tested with chi-square(chi2). The results indicate significant gender difference for tongue thrusting/sucking only (P=0.01) using the fisher's exact test. The relationship between digital sucking habit and mal-occlusion was very significant (P<0.01). PMID- 11922028 TI - Problems with the management of a case of phaeochromocytoma: case report. AB - A 29 year old woman was referred from Jos University Teaching Hospital (J. U.TH.) with a provisional diagnosis of phaeochromocytoma. She had presented with recurrent headaches, palpitations, excessive sweating, dizziness, syncope and hypertension. She was treated with phenoxybenzamine and propranolol for six weeks before the drugs were discontinued. This followed conflicting results of investigations done. The return of persistent and severe symptoms led to recommencement of the drugs and surgical intervention after four weeks. A general anaesthetic technique was used. The surgery was complicated by intraoperative hypertension and hypotension. Phentolamine was used to manage intraoperative hypertension. Postoperative complications included shivering, fatigue and paralytic ileus. The report emphasizes the need to take clinical findings into consideration in interpreting results of investigations. It also illustrates the anaesthetic experience for excision of a phaeochromocytoma. PMID- 11922029 TI - Tuberculosis myocarditis: a case report. AB - A 35-year old man presented with fever, weight loss, drenching night sweats and symptoms of cardiac failure for three months. Examination revealed wasting, peripheral oedema, bilateral pleural effusion and constrictive pericarditis. A diagnosis of constrictive pericarditis with bilateral pleural effusion probably due to tuberculosis was made. Human immunodeficiency virus antibodies and six sputum for acidfast bacilli were negative. Electrocardiograph revealed low voltages globally and echocardiography showed global myocardial hypokinesia. He had pericardiectomy, pericardial and pleural histology was non-specific inflammatory reaction but myocardial histology showed granulomatous changes of tuberculous myocarditis. We suggest that in experienced hands myocardial biopsy could be useful in making the diagnosis. PMID- 11922030 TI - [Immune response in ringworm]. PMID- 11922031 TI - Is one drug enough? Arginine vasopressin in pediatric cardiopulmonary resuscitation. PMID- 11922032 TI - The cost of delaying treatment in multiple sclerosis: what is lost is not regained. PMID- 11922033 TI - Study of the effects of temperature, pH and yeast extract on growth and exopolysaccharides production by Propionibacterium acidi-propionici on milk microfiltrate using a response surface methodology, Journal of Applied Microbiology 2001, 90, 788-796. PMID- 11922034 TI - Product Study. Disposing of the CJD threat. PMID- 11922035 TI - Environmental economics and human health. PMID- 11922036 TI - On the surface: a good idea for killing germs. PMID- 11922037 TI - Problems with the solution. PMID- 11922038 TI - The current of red tide research. PMID- 11922039 TI - Adding another "hatched pink" chromosome. PMID- 11922040 TI - Use of a multiaxial diagnostic system in clinical genetics. PMID- 11922041 TI - Imaging techniques for the myocardial contrast echocardiography. AB - Myocardial contrast echocardiography (MCE) has seen a significant evolution of its clinical application with the advent of newer microbubbles suitable for intravenous administration and development of ultrasound imaging technologies. These new technologies, which are specifically aimed to improve our ability to visualize these microbubbles in the myocardial circulation, require the user to have an understanding of the basis for optimal use in the clinical setting. The discussion below aims to provide a summary of these new microbubble technologies. PMID- 11922042 TI - [Pathophysiological study of severe hearing disorders in children and the countermeasures]. PMID- 11922043 TI - Research news and notes: the pace of medical discovery is accelerating. PMID- 11922044 TI - [History of the local hospital of Rue]. PMID- 11922045 TI - [Private welfare institutions in Gard in 1881]. PMID- 11922046 TI - The chair of dermatovenerology at Zagreb School of Medicine. On occasion of the 80th anniversary of its foundation (1921-2001). AB - The development of dermatovenerology in Croatia parallelled more or less this process in other parts of the Austro-Hungarian Monarchy throughout the 19th century. The foundation of the first specialized hospitals as well as the establishment of dermatovenerological units and their transformation into academic departments and chairs in the early 20th century was a slow growth provoked by the spread of syphilis and Skrljevo disease. A group of Zagreb physicians were instrumental in this regard. Based on archival sources, this paper illustrates the chronology and circumstances of the foundation of the first chair, on the occasion of its 80th anniversary. PMID- 11922047 TI - Multivariate analyses of morphometrical features from Apatemon gracilis (Rudolphi, 1819) Szidat, 1928 and A.annuligerum (v.Nordman, 1832) (Digenea: Strigeidae) metacercariae. AB - Only two species of the strigeid subgenus Apatemon (Apatemon) sensu Sudarikov (1959), Apatemon gracilis (Rudolhi, 1819) and A. annuligerum (von Nordmann, 1832), are known to infect British fishes. A. annuligerum is poorly characterised, with few life-cycle data available, since only the adult and metacercarial stages have been described. Both of these life-stages demonstrate only minor morphological differences from those of A. gracilis. The multidimensional approach of principal components analysis (PCA) was employed to determine the intra- and inter-specific morphological variation that exists for metacercariae of these species. Analyses revealed that metacercarial morphometrics cannot confidently discriminate between Apatemon gracilis and A. annuligerum specimens. Indeed, levels of intra-specific variation present between A. gracilis metacercariae originating from different piscine hosts were as marked for certain populations as those between the two nominal species. Individual populations of A. gracilis metacercariae differed most in the relative dimensions of their internal organs, whereas variation between the two nominal species was primarily in length-related variables. A. gracilis specimens originating from Welsh and Scottish stoneloach represented two populations with the most comparable total body lengths, yet clustered separately. Conversely, A. gracilis metacercariae infecting rainbow trout and salmon parr differed markedly in size, but demonstrated minimal composite morphological variation. Much, if not all, of the variation described between populations and species of parasites can be attributed to host or to metacercarial location within a particular host. These findings and the lack of strong life-cycle data for A. annuligerum place some doubt on its validity as a species discrete from A. gracilis. PMID- 11922048 TI - [Kairos, a Greek diety]. AB - Kairos symbolises both the good opportunity and the inevitable instant. This pretty Greek God took an important part of M. Grmek's life. PMID- 11922049 TI - [Pierre Rayer and the foundation in 1858 of the AGMF: an underrated social political event]. AB - The founding by Rayer in 1858 of the Association Generale des Medecins de France (AGMF), which gathered academic, hospital, and rural or urban doctors, was an act of professional solidarity. This action had a major social and political impact, making obsolete the Le Chapelier law of 1791, which limited to local communities the activity of provident societies. The AGMF also fought against illegal medical acts and for the dignity of medical care. It organized social health care, free for the poor and negotiated for local mutualists, helped establish relations between health care and government, and prepared the 1868 law on industrial injuries. Rayer financed the AGMF from the beginning, and devoted himself to it as a renowned clinician fully aware of the medical applications of science and as the founding President of the Societe de Biologie and of Napoleon III's private physician, who fully supported the AGMF. PMID- 11922050 TI - Universal coverage in the United States: lessons from experience of the 20th century. PMID- 11922051 TI - Drug coverage for Medicare beneficiaries: why protection may be in jeopardy. PMID- 11922052 TI - [The "Diatetic" of Isaac Judaeus]. PMID- 11922053 TI - [The old-french fever tract, fevers]. PMID- 11922054 TI - [An historical review of the use of maggots in wound therapy]. AB - Since William Baers first publication about the use of fly larvae in the treatment of wounds in the year 1931 more than hundred papers have been published about this topic. However, little is known about the first observations that finally led to the widespread use of this technique. The current paper reviews the historic background of the utilization of blowfly-maggots in wound therapy until its decline with the advent of antibiotic drugs and its revival since the eighties of the 20th century. PMID- 11922055 TI - Otolaryngology. PMID- 11922056 TI - [Professor Cheng Zhifan: his academic thinking, scholarship, and behaviour]. PMID- 11922057 TI - Implementation of the Comprehensive Methamphetamine Control Act of 1996; regulation of pseudoephedrine, phenylpropanolamine, and combination ephedrine drug products and reports of certain transactions to nonregulated persons. Final rule. AB - DEA is amending its regulations to implement the requirements of the Comprehensive Methamphetamine Control Act of 1996 (MCA) with respect to the regulation of pseudoephedrine, phenylpropanolamine, and combination ephedrine drug products as List I chemicals, and the reporting of certain transactions involving pseudoephedrine, phenylpropanolamine, and combination ephedrine drug products. The MCA removed the previous exemption from regulation as List I chemicals which had applied to pseudoephedrine, phenylpropanolamine, and combination ephedrine drug products. This action makes persons who distribute the products subject to the registration requirement. Also, distributions, importations, and exportations of the products became subject to the existing chemical controls relating to regulated transactions, except in certain circumstances specified in the MCA. The MCA also requires that reports be submitted for certain distributions involving pseudoephedrine, phenylpropanolamine, and ephedrine (including drug products containing those chemicals) by Postal Service or private or commercial carrier to nonregulated persons. This final rule amends the regulations to make them consistent with the language of the MCA and to establish specific procedures to be followed to satisfy the new reporting requirement. DEA has, where possible, taken action to limit the public impact of these new requirements while remaining consistent with the intent of the MCA to attack the diversion of regulated drug products to the clandestine manufacture of methamphetamine. PMID- 11922059 TI - Attitudes toward cloning: Americans overwhelmingly disapprove of both animal and human cloning. PMID- 11922060 TI - Kinetics of phenol and chlorophenol utilization by Acinetobacter species. AB - Although microbial transformations via cometabolism have been widely observed, the few available kinetic models of cometabolism have not adequately addressed the case of inhibition from both the growth and nongrowth substrates. The present study investigated the degradation kinetics of self-inhibitory growth (phenol) and nongrowth (4-chlorophenol, 4-CP) substrates, present individually and in combination. Specifically, batch experiments were performed using an Acinetobacter isolate growing on phenol alone and with 4-CP present. In addition, batch experiments were also performed to evaluate the transformation of 4-CP by resting, phenol-induced Acinetobacter cultures. The Haldane kinetic model adequately predicted the biodegradation of phenol alone, although a slight discrepancy was noted in cases of higher initial phenol concentrations. Similarly, a Haldane model for substrate utilization was also able to describe the trends in 4-CP transformation by the resting cell cultures. The 4-CP transformation by the Acinetobacter species growing on phenol was modeled using a competitive kinetic model of cometabolism, which included growth and nongrowth substrate inhibition and cross-inhibition terms. Excellent agreement was obtained between the model predictions using experimentally estimated parameter values and the experimental data for the synchronous disappearance of phenol and 4-CP. PMID- 11922061 TI - The chemical species distribution and transformation of polyaluminum silicate chloride coagulant. AB - The chemical species distributions of polyaluminum silicate chloride (PASC) and polyaluminum chloride (PACl) determined by Al-Ferron complexation timed spectrophotometric and 27Al-NMR methods, respectively, have been compared and analyzed. The experimental results show that the species distribution and transformation of PASC are different from those of PACl, due to the interaction of polysilicic acid and hydrolyzed aluminum species. At the same basicity (B), the contents of, Al(b), Al13 and the monomer species Almono (also determined by 27Al-NMR) in PASC are lower than those in PACl, while the contents of Al(c) and the Alother determined by 27Al-NMR in PASC are higher than those in PACl. The differences between PASC and PACl with respect to these species enlarge as the molar ratio of Al/Si in PASC decreases. Further, in PACl the ratio of Al13 to Al(b) closes to 1.0, indicating that the amount of the two fractions are similar. In PASC, however, such an agreement does not exist at the lower B values and Al/Si molar ratios. When the B value and Al/Si molar ratios increase, however, the amount of Al13 and Al(b) species tends to close. The study findings indicate that polysilicic acid can react with hydrolyzed aluminum species to form an aluminum silicate polymer composite and result in the change in species distribution of PASC. PMID- 11922062 TI - Kinetics and mechanism of oxidation of tetrachloroethylene with permanganate. AB - The kinetics, reaction pathways and product distribution of oxidation of tetrachloroethylene (PCE) by potassium permanganate (KMnO4) were studied in phosphate-buffered solutions under constant pH, isothermal, completely mixed and zero headspace conditions. Experimental results indicate that the reaction is first-order with respect to both PCE and KMnO4 and has an activation energy of 9.3+/-0.9 kcal/mol. The second-order rate constant at 20 degrees C is 0.035+/ 0.004 M(-1) s(-1), and is independent of pH and ionic strength (I) over a range of pH 3-10 and I approximately 0-0.2 M, respectively. The PCE-KMnO4 reaction may proceed through further oxidation and/or hydrolysis reaction pathways, greatly influenced by the acidity of the solution, to yield CO2(g), oxalic acid, formic acid and glycolic acid. Under acidic conditions (e.g., pH 3), the further oxidation pathway will dominate and PCE tends to be directly mineralized into CO2 and chloride. Under neutral (e.g., pH 7) and alkaline conditions (e.g., pH 10), the hydroxylation pathway dominates the reaction and PCE is primarily transformed into oxalic acid prior to complete PCE mineralization. Moreover, all chlorine atoms in PCE are rapidly liberated during the reaction and the rate of chloride production is very close to the rate of PCE degradation. PMID- 11922063 TI - Mitigation of chlorpyrifos runoff using constructed wetlands. AB - Constructed wetlands have been proposed as a potential best management practice (BMP) to mitigate effects of pesticide-associated agricultural runoff. Wetland mesocosms (14 m x 59-73 m) were amended with chlorpyrifos to simulate a storm runoff event at concentrations of 73, 147 and 733 microg/l. Water, sediment and plant samples collected weekly for 12 weeks indicated that chlorpyrifos rapidly sorbed to sediment and plant material, with approximately 47-65% of measured chlorpyrifos mass retained within the first 30-36 m of wetland mesocosms. Of the measured mass, approximately 55% and 25% were retained by sediments and plants, respectively. A field-scale evaluation of a constructed wetland's mitigation capability was performed in the Lourens River watershed of Cape Town, South Africa. Results indicate that the wetland was able to retain and considerably decrease the concentration (and hence toxicity) of chlorpyrifos and suspended sediment entering the receiving waterbody (Lourens River). This research provides fundamental answers concerning constructed wetland capabilities that are necessary for constructing field-scale systems within agricultural watersheds. PMID- 11922064 TI - Metal partitioning in products of incineration of municipal solid waste. AB - Metals contained in the waste transfer to the waste incineration products, including flue gas, fly ash, and bottom ash, as different oxide, nitride, carbides, and other phases. Most of the metal-based phases formed in incineration are toxic and their emissions need to be strictly controlled. Therefore, behavior of metal species during incineration must be well understood. Such understanding is possible based on the experimental identification of the metal phases formed in the waste combustion and determination of their concentration in various incineration products. To avoid well-known experimental difficulties of the industrial waste incinerators associated with the poor fuel/conditions reproducibility and limited instrumentation, a 140,000 Btu/h pilot-scale, laboratory burner was constructed, characterized and operated at NJIT. A synthetic fuel representative of the municipal solid waste in the US was formulated and produced in 600-Lb batches. The solid fuel contained Fe and SiO2 as main constituents, and was doped with trace amounts of Al, Ni, Cr, Hg and PbO. Several experiments have been conducted on combustion of the synthetic fuel in the pilot-scale incinerator with varying fuel-air equivalence ratio. Both gaseous and condensed combustion products were sampled and analyzed. Atomic absorption spectroscopy and X-ray diffraction were used to analyze total metal contents and metal containing phases in the incineration products. Thermodynamic equilibrium computations were performed to obtain the adiabatic flame temperature and identify the phases of the metal-containing products formed at the equilibrium conditions. The results of the equilibrium computations performed at the varied fuel/air ratios were compared with the observed experimental results. PMID- 11922065 TI - Screening of physical-chemical methods for removal of organic material, nitrogen and toxicity from low strength landfill leachates. AB - Physical-chemical methods have been suggested for the treatment of low strength municipal landfill leachates. Therefore, applicability of nanofiltration and air stripping were screened in laboratory-scale for the removal of organic matter, ammonia, and toxicity from low strength leachates (NH4-N 74-220 mg/l, chemical oxygen demand (COD) 190-920 mg O2/l, EC50 = 2-17% for Raphidocelis subcapitata). Ozonation was studied as well, but with the emphasis on enhancing biodegradability of leachates. Nanofiltration (25 degrees C) removed 52-66% of COD and 27-50% of ammonia, the latter indicating that ammonia may in part have been present as ammonium salt complexes. Biological pretreatment enhanced the overall COD removal. Air stripping (24 h at pH 11) resulted in 89% and 64% ammonia removal at 20 and 6 degrees C, respectively, the stripping rate remaining below 10 mg N/l h. COD removals of 4-21% were obtained in stripping. Ozonation (20 degrees C) increased the concentration of rapidly biodegradable COD (RBCOD), but the proportion of RBCOD of total COD was still below 20% indicating poor biological treatability. The effect of the different treatments on leachate toxicity was assessed with the Daphnia acute toxicity test (Daphnia magna) and algal growth inhibition test (Raphidcocelis subcapitata). None of the methods was effective in toxicity removal. By way of comparison, treatment in a full-scale biological plant decreased leachate toxicity to half of the initial value. Although leachate toxicity significantly correlated with COD and ammonia in untreated and treated leachate, in some stripping and ozonation experiments toxicity was increased in spite of COD and ammonia removals. PMID- 11922066 TI - Mercury removal, methylmercury formation, and sulfate-reducing bacteria profiles in wetland mesocosms. AB - A pilot-scale model was constructed to determine if a wetland treatment system (WTS) could effectively remove low-level mercury from an outfall located at the Department of Energy's Savannah River Site. Site-specific hydrosoil was planted with giant bulrush, Scirpus californicus, and surface amended with gypsum (CaSO4) prior to investigating the biogeochemical dynamics of sediment-based sulfur and mercury speciation. On average, the pilot WTS decreased total mercury concentrations in the outfall stream by 50%. Transformation of mercury to a more "bioavailable" species, methylmercury, was also observed in the wetland treatment system. Methylmercury formation in the wetland was ascertained with respect to sediment biogeochemistry and S. californicus influences. Differences in sulfate reduction rates (SRRs) were observed between mesocosms that received additional decomposing Scirpus matter and mesocosms that were permitted growth of the submerged macrophyte, Potamogeton pusillus. Relative abundance measurements of sulfate-reducing bacteria (SRB) as characterized using oligonucleotide probes were also noticeably different between the two mesocosms. A positive correlation between increased sulfide, dissolved total mercury, and dissolved methylmercury concentrations was also observed in porewater. The data suggest that soluble mercury-sulfide complexes were formed and contributed, in part, to a slight increase in mercury solubility. Observed increases in methylmercury concentration also suggest that soluble mercury-sulfide complexes represent a significant source of mercury that is "available" for methylation. Finally, a volunteer macrophyte, Potamogeton pusillus, is implicated as having contributed additional suspended particulate matter in surface water that subsequently reduced the pool of dissolved mercury while also providing an environment suitable for demethylation. PMID- 11922067 TI - Photocatalytic reactions of phenanthrene at TiO2/water interfaces. AB - The photocatalytic oxidation of phenanthrene was investigated in aqueous TiO2 suspensions under UV light irradiation. Chemical oxygen demand (COD) measurements, UV-Vis spectrophotometer, IR spectrometer and gas chromatography mass spectrometry (GC-MS) analytical techniques were used to monitor the reaction process. Some factors affecting the photodegradation rate were studied and some aromatic intermediates were detected during the reaction process. Fast and complete mineralization of phenanthrene was achieved in this reaction system. PMID- 11922068 TI - Biodegradation of polylactide in aerobic and anaerobic thermophilic conditions. AB - Biodegradable polymers are designed to resist a number of environmental factors during use, but to be biodegradable under disposal conditions. The biodegradation of polylactide (PLLA) was studied at different elevated temperatures in both aerobic and anaerobic, aquatic and solid state conditions. In the aerobic aquatic headspace test the mineralisation of PLLA was very slow at room temperature, but faster under thermophilic conditions. The clear effect of temperature on the biodegradability of PLLA in the aquatic tests indicates that its polymer structure has to be hydrolysed before microorganisms can utilise it as a nutrient source. At similar elevated temperatures, the biodegradation of PLLA was much faster in anaerobic solid state conditions than in aerobic aquatic conditions. The behaviour of PLLA in the natural composting process was similar to that in the aquatic biodegradation tests, biodegradation starting only after the beginning of the thermophilic phase. These results indicate that PLLA can be considered as a compostable material, being stable during use at mesophilic temperatures, but degrading rapidly during waste disposal in compost or anaerobic treatment facilities. PMID- 11922069 TI - Examination of environmental quality of raw and composting de-inking paper sludge. AB - Paper sludges were traditionally landfilled or burned. Over the years, the use of paper sludges on soils has increased, as well as the concerns about their environmental effects. Therefore, the chemical characterization of paper sludges and their young (immature) compost needed to be investigated, and over 150 inorganic and organic chemicals were analyzed in de-inking paper sludge (DPS). In general, nitrogen, phosphorus and potassium contents were low but variable in raw DPS and its young compost. The contents of arsenic, boron, cadmium, cobalt, chromium, manganese, mercury, molybdenum, nickel, lead, selenium, and zinc were also low and showed low variability. However, the copper contents were above the Canadian compost regulation for unrestricted use and required a follow-up. The fatty- and resin acids, and polycyclic aromatic hydrocarbons were the organic chemicals measured at the highest concentrations. For resinic acids, care should be taken to avoid that leachates reach aquatic life. For polycyclic aromatic hydrocarbons, naphthalene should be followed until soil content reaches 0.1 microg g(-1), the maximum allowed for soil use for agricultural purposes according to Canadian Environmental Quality Guidelines. In young compost, the concentration of these chemical families decreased over time and most compounds were below the detection limits after 24 weeks of composting. In raw DPS, among the phenol, halogenated and monoaromatic hydrocarbons, dioxin and furan, and polychlorinated biphenyl families, most compounds were below the detection limits. The raw DPS and its young compost do not represent a major threat for the environment but can require an environmental follow-up. PMID- 11922070 TI - Feasibility study of photoelectrochemical degradation of methylene blue with three-dimensional electrode-photocatalytic reactor. AB - The photoelectrochemical degradation of methylene blue in aqueous solution was investigated with three-dimensional electrode-photocatalytic reactor. It was found that the methylene blue could be degraded more efficiently by photoelectrochemical process than by photocatalytic oxidation or electrochemical oxidation alone. The decolorization efficiency and COD reduction were 95% and 87% for a photoelectrochemical process, respectively, while they were only 78% and 68% for a single electrochemical process and 89% and 71% for a single photochemical process. The TOC reduction of the former also reached as high as about 81% within a reaction time of 30.0 min. And these degradation reactions conformed to pseudo-first-order kinetics. PMID- 11922071 TI - Degradation of azo dye Procion Red MX-5B by photocatalytic oxidation. AB - The photocatalytic oxidation (PCO) of a monoazo dye Procion Red MX-5B under various physico-chemical conditions was investigated. Degradation of the dye by PCO was enhanced by augmentation in UV intensity, titanium dioxide and hydrogen peroxide concentrations but was inhibited by increase in initial dye concentration. The PCO process was affected by pH in a peculiar way. In the presence of 100 mg/l of TiO2 and the absence of H2O2, the highest reaction rate was observed when the initial pH was 10. With 500 mg/l of TiO2 and 10 mM of H2O2, the reaction was the fastest at initial pH of 3-5. The optimal conditions for the degradation of the dye, at an UV intensity of 17 mW/cm2, were determined to be: TiO2 concentration, 500 mg/l; initial H2O2 concentration, 10 mM; initial pH, 5.0. Monitoring of TOC loss showed that the dye was mineralized by 90% within 80 min under these conditions. Nevertheless, the persistence of a low level of TOC indicated that mineralization was not complete and dead-end product(s) which was (were) resistant to PCO might have accumulated. PMID- 11922072 TI - Hydrogen peroxide photolysis, fenton reagent and photo-fenton for the degradation of nitrophenols: a comparative study. AB - The kinetics of the degradation of seven nitrophenols (2-nitrophenol, 4 nitrophenol, 2,5-dinitrophenol, 2,6-dinitrophenol, 2,4-dinitrophenol, 2-methyl 4,6-dinitrophenol, 4-methyl-2,6-dinitrophenol) with the Fenton reagent, photo Fenton, and hydrogen peroxide photolysis was investigated. The efficiency and operating costs for the studied treatments were evaluated and compared. The Fenton reagent was found to be the most efficient and the cheapest way for the nitrophenols (NPs) degradation. The formation of nitrate as a result of mineralisation of organically bounded nitrogen was observed during the treatment of NPs with studied treatment processes. The degrees of organically bounded nitrogen conversion to nitrate after 90% degradation of NPs with the Fenton, photo-Fenton and hydrogen peroxide photolysis were 51-67%, 85-90%, and 50-60%, respectively. According to Daphnia magna acute toxicity test, the Fenton treatment led to complete detoxification of NPs. PMID- 11922073 TI - Pre-oxidation and coagulation of textile wastewater by the Fenton process. AB - This paper evaluates the Fenton process, involving oxidation and coagulation, for the removal of color and chemical oxygen demand (COD) from synthetic textile wastewater containing polyvinyl alcohol and a reactive dyestuff, R94H. The experimental variables studied include dosages of iron salts and hydrogen peroxide, oxidation time, mixing speed and organic content. The results show that color was removed mainly by Fenton oxidation. The color removal reached a maximum of 90% at a reaction time of 5 min under low dosages of H2O2 and Fe2+. In contrast, the COD was removed primarily by Fenton coagulation, rather than by Fenton oxidation. The ratio of removal efficiency between Fenton process and ferric coagulation was 5.6 for color removal and 1.2 for COD removal. It is concluded that Fenton process for the treatment of textile wastewater favors the removal of color rather than COD. PMID- 11922074 TI - NOM characteristics and treatabilities of ozonation processes. AB - The objectives of this study were intended to evaluate the effects of the characteristics of natural organic matter on the treatabilities of ozonation, coagulation, filtration, and granular activated carbon processes. The ultra violet absorbance (UV254) was used as a surrogate parameter to assess each process in reducing the disinfection by-product formation potential (DBPFP). The results indicate that the DBPFP varies with the sources of water samples and treatment processes, but is closely related to the measurement of UV254/DOC. Coagulation/sedimentation can eliminate large molecular weight organic fractions. Both pre- and post-ozonation processes can reduce some of DBP precursors than the conventional treatment process, and are more reliable for reducing the overall DBPFP. PMID- 11922075 TI - Investigation of retardation effects on the titanium dioxide photodegradation system. AB - The photocatalytic bleaching of dyes using TiO2 has appeared promising in laboratory studies, but little attention has been focused on whether other species such as might be found in wastewater have a deleterious effect on the photobleaching. This study describes the effects that result from the presence of ionic species, organic solvents and humic substances on the photobleaching of several dyes. The manner in which the photobleaching was affected by the presence of adventitious materials was carefully examined with an eye toward elucidating the mechanistic origin of the loss of the photochemical efficiency in the reaction. The overall retardation effects can be attributed to the combination of light attenuation, inhibition and competition effects. PMID- 11922076 TI - Mechanisms of integrin-mediated virus attachment and internalization process. AB - Viruses that propagate within vertebrate hosts have adapted many strategies to infect host cells. One of the first steps in a viral infection is the binding of the virus to cell surface molecules. This interaction between a virus and its receptors plays a key role in the multiplication cycle. Entry of viruses into cells is a complex, multistep process, and for several viruses, cell attachment and internalization are distinct steps. A number of virus receptors have been identified; a common family of viral receptors is the integrin family. Integrins are a widely expressed family of cell adhesion receptors, by which cells attach to extracellular matrices; they also mediate important cell-cell adhesion events. Integrins are involved in a number of tissue remodeling events, including embryogenesis, angiogenesis, wound repair, and bone resorption. In addition, several integrins are used by manyviruses in theirinfectious cycle. Virus integrin interactions maybe more complex than previously thought because several viruses can interact with unique integrin regions or can activate distinct signaling pathways. This article will discuss the strategies devised by many viruses in their integrin-mediated attachment or cell entry. PMID- 11922077 TI - Immune function of thyroid stimulating hormone and receptor. AB - Thyroid stimulating hormone (TSH) is a central component of the hypothalamus pituitary-thyroid axis. Although TSH is known for its important biological effects as a neuroendocrine used to regulate thyroid hormone activity and subsequent metabolic functions, TSH also has been shown to be produced and used by cells ofthe mammalian immune system. Moreover, recent findings have linked the use of TSH by cells of the immune system in humans and mice to a group of monocytic cells and lymphocytes--primarily dendritic cells, macrophages, and subset of naive peripheral T cells. Other studies have demonstrated the capacity of dendritic cells and monocytes to produce biologically active TSH, thereby pointing to a process of paracrine or autocrine TSH-mediated communication during the earliest stages of an immune response to antigen. In this article, these and other features of TSH immune-endocrine interactions are discussed in the context of an intrinsic TSH immunological pathway. Additionally, a hypothesis is proposed in which TSH produced by cells of the immune system during acute antigen exposure plays a dual role, consisting on the one hand of TSH communication during antigen driven immune activation while concomitantly serving to regulate physiological homeostasis by modulating and adjusting thyroid hormone activity. PMID- 11922079 TI - Effects of ultraviolet exposure on the immune system. AB - Depletion of stratospheric ozone and changes in lifestyle lead to an increased exposure to ultraviolet (UV) wavebands, especially in the UVB region (280-320 nm). Besides the beneficial effects of UV exposure, such as vitamin D production, cosmetic tanning, and adaptation to solar UV, UV exposure can also have adverse consequences on human health, notably sunburn, skin cancer, and ocular damage. Over the last two and a half decades it has become evident that especially UVB exposure and to a lesser extent UVA modulates specific as well as nonspecific immune responses. Several reports have shown that this immunomodulation plays at least a partial role in the induction of skin cancer. In addition, UVB exposure has been demonstrated to impair resistance to some infections. On the other hand, immunomodulation resulting from UVB exposure might be physiologically important in inhibiting responses to neoantigens in the skin induced by UV exposure. In the last 20 years UV has been used frequently as an experimental tool to unravel immune responses-especially immune responses initiated in the skin (i.e., photoimmunology). In this review, the major mechanisms responsible for UV-induced immunomodulation and its consequences are summarized. PMID- 11922078 TI - Structural plasticity of the proteasome and its function in antigen processing. AB - The proteasome is the main provider of peptide ligands for major histocompatibility complex class I molecules. During an immune response to pathogens, the proinflammatory cytokine interferon (IFN)-gamma and tumor necrosis factor (TNF)-alpha are released, which induce the proteasome subunits LMP2, LMP7, and MECL-1. These replace the constitutively expressed active site subunits of the proteasome (delta, MB1, and Z) leading to a marked change in the cleavage preference of the proteasome and the production of T-cell epitopes. Proteasome activity is further changed by the IFN-gamma-mediated induction of the proteasome regulator PA28alpha/beta and the downregulation of PA28gamma. Why such an extensive exchange of proteasome active site subunits and regulators occurs is still poorly understood. In this article we discuss recent insights in the structural consequences of proteasome reorganization and their effects on epitope generation and shaping of the cytotoxic immune response. Moreover, we review the latest data on how the ubiquitin pathway targets protein antigens for peptide processing and discuss the potential of proteasome inhibitors for the modulation of antigen presentation. PMID- 11922080 TI - Recent advances in analytical methodology for cyclopiazonic acid. AB - Cyclopiazonic acid (CPA) is a toxic indole tetramic acid that has been isolated from numerous species of Aspergillus and Penicillium. It has been found as a natural contaminant of cheese, corn, peanuts and various feedstuffs. Historically, thin-layer chromatography has been the most widely used method for quantitative determination of CPA in fungal cultures and agricultural commodities. Several liquid chromatographic (LC) and spectrophotometric methods have also been used, but these require extensive, time-consuming cleanup procedures to achieve accurate quantitation. More recently, enzyme-linked immunosorbent assays (ELISA) have been developed for quantification of CPA, and an immunoaffinity column (IAC) has been developed for cleanup of sample extracts prior to quantification by ELISA or LC. In applying the IAC to the cleanup of peanut extracts, recovery of CPA from spiked samples ranged from 83.7% to 90.8%, and the method was successfully applied to the analysis of peanuts that were naturally contaminated with CPA. PMID- 11922081 TI - Methods of analysis for ochratoxin A. AB - The mycotoxin ochratoxin A (OTA) is produced by the fungi Aspergillus alutaceus and Penicillium verrucosum and has carcinogenic, nephrotoxic, teratogenic and immunosuppressive properties. The levels of OTA in foodstuffs are regulated in several countries, so reliable and sensitive methods are necessary for its determination. Procedures for extraction of OTA from ground foods generally use an organic solvent in the presence of acid or an extraction solvent containing aqueous sodium bicarbonate. Cleanup procedures include partition into aqueous sodium bicarbonate, solid phase extraction (SPE) columns and immunoaffinity chromatography. The latter technique allows detection of sub-ppb levels of OTA in a wide variety of foods and in plasma. The most widely used determinative procedure is reversed phase liquid chromatography (LC) with detection by fluorescence (excitation 330-340 nm, emission 460-470 nm) or, more recently, by tandem mass spectrometry. ELISA methods are also available. Certified reference materials containing OTA have been prepared. PMID- 11922082 TI - Methods for the determination of deoxynivalenol and other trichothecenes in foods. AB - Trichothecene mycotoxins are secondary metabolites of Fusarium moulds that routinely infect cereal crops. Processing can reduce, but not eliminate, trichothecenes from cereal-based foods, and the potential presence of the trichothecenes in cereal foods poses a significant health risk to consumers. Deoxynivalenol (vomitoxin, DON) is the most common of the trichothecenes detected in cereal crops and is subject to government regulation in many countries. Sensitive (ng/g) methods for the detection of trichothecenes in cereal grains and food products are needed to protect consumers, to provide data for dietary exposure estimates, and to support research into the control of moulds and subsequent toxin production. Laboratories require simple, rugged and reliable methods for routine testing, with unequivocal identification of suspect mycotoxins. A method employing gas chromatography-negative ion chemical ionisation/mass spectrometry (GC-NICI/MS) has been developed and used for the routine determination of eight of the most significant trichothecenes in a variety of commodities. This chapter discusses GC, liquid chromatographic (LC) and supercritical fluid chromatographic methods that are currently used for the analysis of trichothecene mycotoxins. PMID- 11922083 TI - Characterization of clay-based enterosorbents for the prevention of aflatoxicosis. AB - Appropriate chemical interventions that can block, retard, or significantly diminish foodborne exposure to aflatoxins are high priorities. A practical and effective approach to the aflatoxin problem has been the dietary inclusion of a processed calcium montmorillonite clay (HSCAS). HSCAS acts as an enterosorbent that rapidly and preferentially binds aflatoxins in the gastrointestinal tract resulting in decreased aflatoxin uptake and bioavailability. In mechanistic studies, we have shown that the intact dicarbonyl system in aflatoxin is essential for optimal sorption by HSCAS. Evidence also suggests that aflatoxins react at multiple sites on HSCAS clay surfaces (especially those within the interlayer region). Due to conceivable risks associated with the dietary inclusion of nonspecific binding agents, all aflatoxin enterosorbents should be tested in sensitive animal models for efficacy, safety, and the potential for nutrient interactions. PMID- 11922084 TI - Effect of processing on aflatoxin. AB - Naturally occurring toxicant contamination of foods with mycotoxins is unavoidable and unpredictable and poses a unique challenge to food safety. Aflatoxins are toxic mold metabolites produced by toxigenic strains of Aspergillus species. Primary commodities susceptible to aflatoxin contamination include corn, peanuts and cottonseed and animal-derived foods such as milk when the animal is fed aflatoxin-contaminated feed. Risks associated with aflatoxin contaminated foods can be reduced through the use of specific processing and decontamination procedures. Factors, which influence the effectiveness of a specific process or procedure, include the chemical stability of the mycotoxin(s), nature of the process, type and interaction with the food/feed matrix and interaction with multiple mycotoxins if present. Practical decontamination procedures must: 1) inactivate, destroy, or remove the toxin, 2) not produce or leave toxic residues in the food/feed, 3) retain the nutritive value of the food/feed, 4) not alter the acceptability or the technological properties of the product, and, if possible, 5) destroy fungal spores. For aflatoxins, multiple processing and/or decontamination schemes have been successful in reducing aflatoxin concentrations to acceptable levels. Physical cleaning and separation procedures, where the mold-damaged kernel/seed/nut is removed from the intact commodity, can result in 40-80% reduction in aflatoxins levels. Processes such as dry and wet milling result in the distribution of aflatoxin residues into less utilized fractions of the commodity. The ammoniation of aflatoxin-contaminated commodities has altered the concentrations as well as toxic and carcinogenic effects of aflatoxin by greater than 99%. Nonbiological materials such as selected anticaking agents covalently bind aflatoxins from aqueous suspensions, diminish aflatoxin uptake by animals, prevent acute aflatoxicosis, and decrease aflatoxin residues in milk. Ultimately, the best processing or decontamination process is one that is approved by regulatory agencies, cost-effective, and reduces the mycotoxin concentration to acceptable levels. PMID- 11922085 TI - Effect of processing on deoxynivalenol and other trichothecenes. AB - Deoxynivalenol (DON, vomitoxin) and other trichothecene toxins may contaminate crops as weather conditions during the growing season sometimes favor the growth and toxin production by the Fusarium fungus. Several processing procedures for the reduction of DON in contaminated wheat and corn have been studied. Although total elimination of toxin is not usually possible, processing methods such as cleaning and milling can reduce toxin to acceptable levels. In addition, more countries have guidelines or advisory levels to follow for the utilization of DON contaminated grains such as wheat. In recent years, a few studies on grain processing have been conducted, however, there is a continuing effort to find a more effective method or a combination of methods for the reduction of DON in grains and grain products. PMID- 11922086 TI - Effect of processing on ochratoxin A (OTA) content of coffee. AB - Coffee production can be roughly separated into three main steps 1) cherry processing to green coffee beans, 2) storage and transportation of green coffee to the place of consumption, and 3) green coffee processing to roasted and ground coffee and soluble coffee. The mold species which are known to produce ochratoxin A (OTA) in coffee have been identified as Aspergillus ochraceus, A. carbonarius and occasionally, A. niger. The length of time spent at a water activity > 0.80 at any moment until roasting defines the risk of mold growth and OTA production. However, the specific moment and locus of contamination have not yet been clearly identified. Since coffee husks are a significant source of OTA contamination, cleaning and grading of green coffee are effective methods for reducing OTA levels in coffee. During the process of converting green coffee to roasted and soluble coffees, up to 90% reduction in OTA levels can occur. PMID- 11922087 TI - Aspects of the ecology of Fusarium toxins in cereals. AB - Species of the genus Fusarium account for three of the five agriculturally important mycotoxins which are deoxynivalenol, aflatoxin, fumonisin, zearalenone and ochratoxin. The toxigenic fusaria have been complicated to study because morphologically-similar strains represent different biologies: saprophytes, pathyotypes and endophytes. This might explain the difficulties with systems of taxonomy for Fusarium species and increasing reliance on molecular techniques to characterize taxa. Another remarkable feature of the toxigenic fusaria is that each species produces compounds that cross several species as well as families of compounds that are species specific. In addition, reproductively-isolated strains (from different continents) of important species such as F. graminearum produce different compounds, and even produce the same compounds by different biosynthetic pathways. PMID- 11922088 TI - Stability of fumonisins in food processing. AB - Fumonisins are mycotoxins produced by Fusarium verticillioides (moniliforme) and Fusarium proliferatum that are found in corn and processed corn-based food products. Although generally heat stable, fumonisin concentrations appear to decline as processing temperatures increase. At processing temperatures of 125 degrees C or lower, losses of fumonisin are low (25-30%), whereas at temperatures of 175 degrees C and higher, losses are greater (90% or more). Processes such as baking and canning, where product temperatures rarely reach 175 degrees C, result in little or no loss of fumonisin. Processes such as frying and extrusion cooking, where temperatures can exceed 175 degrees C, result in greater losses. Heating fumonisin in the presence of glucose results in an apparent first order loss of the toxin. Adding glucose to corn muffins and extrusion mixes results in high losses of fumonisins during baking and extrusion processing. Little information exists on the effects of chemical and bioprocessing on fumonisins. Alkaline processing of corn, such as in the nixtamalization process, hydrolyzes fumonisins and results in a more toxic product. Additional research is needed to identify and to determine the toxicity of fumonisin decomposition products. PMID- 11922089 TI - Effects of processing on zearalenone. AB - Zearalenone (ZEN), a common contaminant of all major cereal grains worldwide, is produced by some plant pathogenic molds including Fusarium graminearum and F. culmorum. The biological activity of this mycotoxin is mainly attributed to its estrogenic activity that modulates/disrupts endocrine function in animals and possibly humans. Efforts have been made to reduce the level of ZEN by various chemical, physical, and biological processing methods. Some chemical treatments were shown to be effective in reducing zearalenone content in artificially or naturally contaminated foods. During physical processing, the fate of ZEN depended on its distribution in the food matrix and its chemical properties such as heat stability and solubility. For example, wet milling of contaminated corn resulted in starch that was essentially toxin-free. In contrast, animal feed fractions such as bran and germ, by-products of the wet milling process, tended to concentrate ZEN. Extrusion cooking, a complex process where food is subjected to heat, high pressures and shear stress, reduced ZEN levels in food as well as its estrogenic activity. Fermentation of foods with bacteria and yeast resulted in reduction in ZEN levels. However, fermentation can result in the conversion of ZEN to more potent derivatives such as cc-zearalenol. Further efforts are needed to identify effective methods for removing/detoxifying ZEN in foods. PMID- 11922090 TI - Mycotoxins and fermentation--beer production. AB - Along with food safety issues due to mycotoxins, the effects of Fusarium infections on malt and beer quality can be disastrous. While some of the Fusarium head blight mycotoxins, such as DON, present in infected barley may be lost during steeping, the Fusarium mold is still capable of growth and mycotoxin production during steeping, germination and kilning. Therefore, detoxification of grain before malting may not be practical unless further growth of the mold is also prevented. Methods for reducing the amount of mold growth during malting are needed. Physical, chemical and biological methods exist for inhibiting mold growth in grain. Irradiation is a promising means for preventing Fusarium growth during malting, but its effects on malt quality and mycotoxin production in surviving mold need to be evaluated in more detail. Chemical treatments such as ozonation, which do not leave chemical residues in beer, also appear to be promising. Although biological control methods may be desirable, the effects of these inhibitors on malt and beer quality require further investigation. In addition, storage studies are needed to determine the effect of biological control on Fusarium viability and malt quality. It may also be possible to incorporate detoxifying genes into fermentation yeasts, which would result in detoxification of mycotoxins present in wort. Development of these types of technological interventions should help improve the safety of products, such as beer, made from Fusarium infected grain. PMID- 11922091 TI - Aflatoxin, hepatitis and worldwide liver cancer risks. AB - Aflatoxins are among the most potent mutagenic and carcinogenic substances known. Differential potency of aflatoxin among species can be partially attributed to differences in metabolism; however, current information on competing aspects of metabolic activation and detoxification of aflatoxin in various species does not identify an adequate animal model for humans. Risk of liver cancer is influenced by a number of factors, most notably carriage of hepatitis B virus as determined by the presence in serum of the hepatitis B surface antigen (HBsAg+ or HBsAg-). About 50 to 100% of liver cancer cases are estimated to be associated with persistent infection of hepatitis B (or C) virus. The potency of aflatoxin in HBsAg+ individuals is substantially higher (about a factor of 30) than the potency in HBsAg- individuals. Thus, reduction of the intake of aflatoxins in populations with a high prevalence of HBsAg+ individuals will have greater impact on reducing liver cancer rates than reductions in populations with a low prevalence of HbsAg+ individuals. The present analysis suggests that vaccination against hepatitis B (or protection against hepatits C), which reduces prevalence of carriers, would reduce the potency of the aflatoxins in vaccinated populations and reduce liver cancer risk. PMID- 11922093 TI - Worldwide regulations for mycotoxins. AB - Since the discovery of the aflatoxins in the 1960s, regulations have been established in many countries to protect the consumer from the harmful effects of mycotoxins that may contaminate foodstuffs. Various factors play a role in the decision-making process of setting limits for mycotoxins. These include scientific factors such as the availability of toxicological data, survey data, knowledge about the distribution of mycotoxins in commodities, and analytical methodology. Economical and political factors such as commercial interests and sufficiency of food supply have their impact as well. International enquiry's on existing mycotoxin legislation in foodstuffs and animal feedstuffs have been carried out several times in the 1980s and 1990s and details about tolerances, legal basis, responsible authorities, official protocols of analysis and sampling have been published. Recently a comprehensive update on worldwide regulations was published as FAO Food and Nutrition Paper 64. It appeared that at least 77 countries now have specific regulations for mycotoxins, 13 countries are known to have no specific regulations, whereas no data are available for about 50 countries, many of them in Africa. Over the years, a large diversity in tolerance levels for mycotoxins has remained. Some free trade zones (EU, MERCOSUR) are in the process of harmonizing the limits and regulations for mycotoxins in their respective member states, but it is not likely that worldwide harmonized limits for mycotoxins will soon be within reach. PMID- 11922092 TI - Risk assessment of ochratoxin: current views of the European Scientific Committee on Food, the JECFA and the Codex Committee on Food Additives and Contaminants. AB - The chlorinated isocoumarin compound, ochratoxin A (OTA), together with some related derivatives (ochratoxins B, C, alpha, beta) are produced by Penicillium verrucosum and by several spp. of Aspergillus, most notably A. ochraceus. P. verrucosum is the principal source of OTA contamination of stored foods in temperate climates while Aspergillus spp. predominate in warmer countries. The major dietary sources of OTA are cereals but significant levels of contamination may be found in grape juice and red wine, coffee, cocoa, nuts, spices and dried fruits. Because of the chemical stability of OTA and long half-life in mammalian tissues, contamination may also carry over into pork and pig blood products and into beer. OTA is potently nephrotoxic and carcinogenic, the potency varying markedly between species and sexes; it is also teratogenic and immunotoxic. There have been different approaches to the risk assessment of OTA in different jurisdictions, largely arising from whether or not the carcinogenicity of OTA is considered to arise through a thresholded or non-thresholded mechanism. Consequently the tolerable intakes have variously been estimated at 100 ng/kg bw/week (JECFA), 1.5 to 5.7 ng/kg bw/day (Canada) and not more than 5 ng/kg bw/day (European Commission). These differences are also reflected in risk management measures that have been implemented or proposed with different maximum contamination levels being applied to different commodities and to the same commodity in different countries. Prevention of contamination at source is considered to be the most effective public health measure. There is also a need to harmonise the risk assessment and management processes to a greater extent than currently exist if barriers to trade are to be avoided. PMID- 11922094 TI - Economic changes imposed by mycotoxins in food grains: case study of deoxynivalenol in winter wheat. AB - The Fusarium epidemic of 1996 in Ontario winter wheat resulted in direct losses of well over $100 million Canadian dollars (CDN). More importantly, wheat marketing in Ontario has changed. The market focus remains primarily food grade. Thus, the awareness of deoxynivalenol (DON) entering the food chain has influenced marketing and trade. New market tolerances for DON have been set. The Chicago Board of Trade will only accept up to 5 ppm (DON), while a new tolerance of 0.5 ppm DON has been set in the breakfast cereal markets. Advance contracts are avoided and there is a new reluctance to service export customers, all because of the liability associated with DON. Furthermore, there are no markets for process by-products which contain concentrated levels of DON. Before 1996, DON problems were handled largely by blending grain across the province. A reluctance to blend DON contaminated wheat and site specific sourcing of grain is growing. PMID- 11922095 TI - U.S. perspective on mycotoxin regulatory issues. AB - Control programs set up by the Food and Drug Administration (FDA) for aflatoxin, an unavoidable natural contaminant produced by specific molds that invade a number of feedstuffs and basic foods, provide an example of forces that affect risk assessment and management strategies by a regulatory agency. More recently, on an international scale, efforts to establish international food standards for fumonisin, deoxynivalenol, ochratoxin A, zearalenone, and patulin, as well as for aflatoxin, demonstrate the complexity of developing regulations and/or standards designed to protect consumer health and ensure fair trade practices on a global scale. Current FDA regulations for aflatoxins address public health concerns for potential contamination in basic foods, residues in milk, and animal feeds for numerous commodities and applications. Regulatory limits, sampling and analytical procedures, decontamination and/or diversion to less risk uses for contaminated product are components of mycotoxin control programs. Current efforts by FDA to establish regulatory controls for deoxynivalenol, fumonisin, and patulin add further insight on the role that safety and risk assessment procedures play in the development of action levels and advisories for mycotoxins. PMID- 11922096 TI - Biology and ecology of toxigenic Penicillium species. AB - Many Penicillium species produce mycotoxins. The importance of these toxic compounds varies widely, and is governed as much by the biology and ecology of the species concerned as by the inherent toxicity of the compounds themselves. For example, P. citreonigrum and P. islandicum make potent toxins, but as both species are rare in nature, the toxins are not important. Although P. janthinellum and P. simplicissimum are very widely distributed and make potent toxins, these species are rarely found outside soils so again, the toxins are of little practical importance. The very common P. crustosum makes a potent tremorgenic mycotoxin, fortunately, the toxin is only produced at very high water activities. On the other hand, P. verrucosum, unknown in the tropics, is widespread in cereals in cold climates. Consequently, ochratoxin A production by this species causes a major toxicosis. The biology and ecology of these and other Penicillium mycotoxins will be described in this paper. PMID- 11922097 TI - Biology and ecology of mycotoxigenic Aspergillus species as related to economic and health concerns. AB - The fungal genus Aspergillus was established in 1729, and includes species that are adapted to a wide range of environmental conditions. Many aspergilli produce mycotoxins in foods that may be toxic, mutagenic or carcinogenic in animals. Most of the Aspergillus species are soil fungi or saprophytes but some are capable of causing decay in storage, disease in plants or invasive disease in humans and animals. Major agricultural commodities affected before or after harvest by fungal growth and mycotoxins include corn, peanuts, cottonseed, rice, tree nuts, cereal grains, and fruits. Animal products (meat, milk and eggs) can become contaminated because of diet. Aspergillus flavus, A. parasiticus, A. ochraceus, A. niger, A. fumigatus and other aspergilli produce mycotoxins of concern. These include the aflatoxins and ochratoxins, as well as cyclopiazonic acid, patulin, sterigmatocystin, gliotoxin, citrinin and other potentially toxic metabolites. PMID- 11922098 TI - Chemistry and toxicology of molds isolated from water-damaged buildings. AB - There is increasing evidence of health risks associated with damp buildings and homes in which high levels of microbes are found. Although concerns have traditionally centered on microbial pathogens and allergenic effects, recent work has suggested that fungi pose the more serious risk. Evidence is accumulating that certain toxigenic molds are particularly a risk for human health through exposure, via inhalation, of fungal spores. Many of these fungi produce toxins (mycotoxins) some of which have been shown to cause animal and human intoxications, usually in an agricultural setting. The fungus, Stachybotrys chartarum (S. atra) is considered to be one of the more serious threats to people living and working in water-damaged buildings. This mold has a long history of being responsible for animal toxicoses, and in recent years, being associated with infant pulmonary hemosiderosis (bleeding in the lungs) of infants exposed to spores of this fungus in their homes. S. atra produces a variety of potent toxins and immunosuppressant agents, including a novel class of diterpenes (atranones) of unusual structure. More research is needed to determine the impact to health resulting from inhalation of toxigenic mold spores. PMID- 11922099 TI - Biological control of Fusarium head blight of wheat and deoxynivalenol levels in grain via use of microbial antagonists. AB - Efforts to reduce mycotoxin contamination in food logically start with minimizing plant infection by mycotoxin producing pathogens. Fusarium graminearum (perfect state, Gibberella zeae) infects wheat heads at flowering, causing the disease Fusarium head blight (FHB) and losses of over 2.6 billion dollars in the U.S. during the last 10 years. The pathogen often produces deoxynivalenol (DON) resulting in grain size and quality reduction. Highly resistant wheat cultivars currently are not available for reducing FHB, and labeled fungicides are not consistently effective. The feasibility of biologically controlling FHB is currently being evaluated. Microbial isolates obtained from wheat anthers were screened for their ability to utilize tartaric acid, a compound that is poorly utilized by F. graminearum and could be utilized in formulations of biological control agents. Four strains that utilized tartaric acid and three that did not were effective in reducing FHB disease severity by up to 95% in greenhouse and 56% in field trials. Additional research programs around the globe have identified other antagonist strains with potential for biologically controlling FHB. Though a considerable body of research remains to be completed, strategies and microorganisms for biologically controlling FHB have reached an advanced stage of development and offer the promise of being an effective tool that could soon contribute to the reduction of FHB severity and DON contamination of grain in commercial agriculture. PMID- 11922101 TI - Novel assays and sensor platforms for the detection of aflatoxins. AB - The importance of the aflatoxins from food safety and economic standpoints has continued to drive the development of new analytical methods for these mycotoxins. Currently the widely used methods for measurement of aflatoxins fall into two groups, the established chromatographic methods and traditional enzyme linked immunosorbent assays (ELISAs). Recently substantial progress has been made in the application of new technologies to the monitoring of aflatoxins. In particular, several research groups have developed biosensors for detection ofthe toxins as well as presumptive tests for fungal infection. Biosensors have been developed in a variety of formats including surface plasmon resonance, fiber optic probes, and microbead-based assays. The sensitivity and selectivity of the biosensors and of the presumptive tests has reached the level the where the application of these techniques to the screening of foods warrants further investigation. PMID- 11922100 TI - Sampling wheat for deoxynivalenol. AB - The variability associated with testing wheat for deoxynivalenol (DON) was measured using a 0.454 kg sample, a Romer mill, 25 g of comminuted subsample and the Romer Fluoroquant analytical method. The total variability was partitioned into sampling, sample preparation, and analytical variability components. Each variance component was found to be a function of the DON concentration and equations were developed to predict each variance component using regression techniques. The effects of sample size, subsample size, and number of aliquots on reducing the variability of the DON test procedure were also determined. Using the test procedure described above, the coefficient of variation (CV) associated with testing wheat at 5 ppm DON was found to be 13.4%. The CVs associated with sampling, sample preparation, and analysis were 6.3, 10.0, and 6.3%, respectively. The sample variations associated with testing wheat are relatively small when compared to CVs associated with testing other commodities for other mycotoxins such as aflatoxin in peanuts. Even with the use of a small sample size (0.454 kg), the sampling variation was not the largest source of error as found in other mycotoxin test procedures. PMID- 11922102 TI - Electrospray mass spectrometry for fumonisin detection and method validation. AB - Fumonisins are a structurally related group of mycotoxins, characterized by a 19 20 carbon aminopolyhydroxy-alkyl chain which is diesterified with propane-1,2,3 tricarboxylic acid (tricarballylic acid). These mycotoxins are commonly found in corn and corn-based food products and have been linked to a variety of animal toxicities. The widespread prevalence of fumonisins and the toxicity associated with ingestion has resulted in a number of analytical methods for determining the amount of fumonisins present in foods. Among the most common of these methods are liquid chromatographic (LC) separation with fluorescence detection, enzyme-linked immunosorbent assay (ELISA) and LC/mass spectrometry. LC and ELISA give quantitative results while LC/MS provide quantitative analysis as well as confirmation of identity of the fumonisins. PMID- 11922103 TI - A system of complementary genes in hybrids between Drosophila koepferae and D. buzzatii: a Markov chain model allows to make inferences about their number and relationships. AB - In backcrosses between D. koepferae and D. buzzatii, the disruption of a system of species-specific complementary factors brings about hybrid male inviability. This system consists of a lethal factor, hmi-1, linked to the X chromosome of D. koepferae, and several conspecific autosomal suppressors. However, hmi-1 hybrid males can also be rescued by factors present in some strains of D. buzzatii. The present work aims to estimate the number of hmi-1 suppressors in one of these strains by means of Markov chains. The obtained results allow discarding models with one or more chromosomes having independent suppressor effect. On the other hand, models having n chromosomes that interact in groups of r, being 1 < r < or = n, to produce rescue effect, provide good approximations to the observed results. The best fit to the data is obtained with four or five chromosomes with suppressor effect, interacting epistatically in groups of three to rescue the viability of hmi-1 males. PMID- 11922104 TI - Molecular evolutionary analysis of a histone gene repeating unit from Drosophila simulans. AB - A repeating unit of the histone gene cluster from Drosophila simulans containing the H1, H2A, H2B and H4 genes (the H3 gene region has already been analyzed) was cloned and analyzed. A nucleotide sequence of about 4.6 kbp was determined to study the nucleotide divergence and molecular evolution of the histone gene cluster. Comparison of the structure and nucleotide sequence with those of Drosophila melanogaster showed that the four histone genes were located at identical positions and in the same directions. The proportion of different nucleotide sites was 6.3% in total. The amino acid sequence of H1 was divergent, with a 5.1% difference. However, no amino acid change has been observed for the other three histone proteins. Analysis of the GC contents and the base substitution patterns in the two lineages, D. melanogaster and D. simulans, with a common ancestor showed the following. 1) A strong negative correlation was found between the GC content and the nucleotide divergence in the whole repeating unit. 2) The mode of molecular evolution previously found for the H3 gene was also observed for the whole repeating unit of histone genes; the nucleotide substitutions were stationary in the 3' and spacer regions, and there was a directional change of the codon usage to the AT-rich codons. 3) No distinct difference in the mode or pattern of molecular evolution was detected for the histone gene repeating unit in the D. melanogaster and D. simulans lineages. These results suggest that selectional pressure for the coding regions of histones, which eliminate A and T, is less effective in the D. melanogaster and D. simulans lineages than in the other GC-rich species. PMID- 11922105 TI - Genome analysis of Agrobacterium tumefaciens: linkage map and genetic features of the left region of the linear chromosome. AB - In addition to a unique tumor-inducing (Ti) plasmid, the plant pathogenic bacterium Agrobacterium tumefaciens has an unconventional chromosomal organization. Our previous studies on A. tumefaciens MAFF301001 revealed that it possesses a 2 Mb linear and a 2.8 Mb circular chromosome plus a 206.479 kbp Ti plasmid (pTi-SAKURA). In this study, a linkage map for the left half of its linear chromosome covering a 900 kbp region was constructed and the number of potential genes existing in the region was estimated. The linkage map consists of 31 BAC and 8 lambda phage recombinants without any gaps. It confirmed the size and all the structural landmarks indicated in the corresponding region of our previously constructed physical map for the linear chromosome. Sequencing analysis of the end-regions of each linking clone led to the identification of 6 genes and another 27 potential genes or ORFs, including genes and/or gene clusters responsible for homologous recombination (ruvB), trehalose/maltose sugar transport (thuR, thuG) and alanine catabolism (dadR). Two virulence-related gene homologues (attK and celB), previously reported in the circular chromosome of a different strain of A. tumefaciens were found in this region. These findings will provide a ready-to-use linkage map for further functional analysis of the linear chromosome. PMID- 11922106 TI - Auxin response factor family in rice. AB - We isolated 11 rice genes homologous to the genes encoding auxin response factors (ARFs) in Arabidopsis. All of the genes encoded a well-conserved amino acid sequence in the N-terminal region, which is considered to be a DNA-binding domain (DBD). Phylogenetic analysis based on comparison of the DBDs indicated that rice has one or two closely related orthologs corresponding to a given respective ARF gene in Arabidopsis. We also analyzed the amino acid sequences of another conserved domain in the C-terminal conserved domain (CTD), which was shared by almost all the rice ARFs, with the exception of OsETTIN1 and OsETTIN2. These results agreed well with the evolutionary relationship deduced from the DBD comparison. In contrast to many ARFs, OsETTIN1 and OsETTIN2 do not contain the conserved C-terminal domain, but do share another consensus motif that is also found in Arabidopsis ETTIN. All of the above observations indicate that rice has functionally diversified ARF genes whose structures and functions correspond to those of various Arabidopsis ARFs, with one or two rice ARFs corresponding to a given Arabidopsis ARF. Thus, auxin signal transduction mechanisms may be well conserved between monocot and dicot plants. PMID- 11922107 TI - Mutations that cause amino acid substitutions at the invariant positions in homeodomain of OSH3 KNOX protein suggest artificial selection during rice domestication. AB - KNOX homeodomain (HD) proteins encoded by KNOTTED1-like homeobox genes (KNOX genes) are considered to work as important regulators for plant developmental and morphogenetic events. We found that OSH3, one of the KNOX genes isolated from a cultivar of Oryza sativa (Nipponbare), encodes a novel HD, which has two amino acid substitutions at invariant positions. Sequence analysis of OSH3 from various domesticated and wild species of rice has revealed that these substitutions are distributed only in Japonica and Javanica type of O. sativa, two groups of domesticated rice in Asia. Surprisingly, nucleotide sequences in the first intron are almost conserved in the rice strains that have the substitutions at the invariant amino acids. Overexpression studies revealed that these invariant amino acids are critical for the function of OSH3 in vivo. The facts that these substitutions occurred specifically at the functionally important amino acids and the sequences are conserved in intron where neutral mutations accumulate suggest the substitutions at the invariant positions of OSH3 have been fixed by artificial selections during domestication. Based on these observations, we hypothesize that OSH3 is responsible for one of the traits that are selectively introduced during the domestication of most of Japonica and a part of Javanica type of rice. PMID- 11922108 TI - Defects in glycosylphosphatidylinositol (GPI) anchor synthesis activate Hog1 kinase and confer copper-resistance in Saccharomyces cerevisisae. AB - Las21/Gpi7 contains a heavy-metal-associated motif at its N-terminus. When this motif was disrupted by amino acid substitution, the cells acquired weak copper resistance. We found that the previously isolated las21 mutants were strongly resistant to copper. Metallothionein is necessary for the expression of the copper-resistance of the las21 mutants. However, hyper-production of metallothionein is unlikely to be the cause of copper-resistance of the las21 mutants. Copper-sensitive mutants (collectively called Cus mutants) were isolated from the las21delta and characterized. One of the Cus genes was found to be PBS2, which encodes Hog1 MAP kinase kinase, indicating that the Hog1 MAP kinase pathway is needed for the expression of copper-resistance of the las21 mutants. As expected, the las21delta hog1delta strain was no longer copper-resistant. We found that Hog1 was constitutively activated in las21delta cells and in ssk1delta las21delta cells but not in sho1delta las21delta cells. Inactivation of either FSR2/MCD4 or MPC1/GPI13, both of which are involved in GPI anchor synthesis, like LAS21, caused a similar level of constitutive activation of Hog1 kinase and copper-resistance as found in the las21delta strain. The constitutive activation was canceled by introducing the sskl mutation, but not the sho1 mutation, in each GPI anchor mutant tested, suggesting that the defect in GPI anchor synthesis specifically affects the Slnl branch of the MAP kinase pathway. Since the wild type cells grown in YPD containing 0.5 M NaCl do not show copper-resistance, mere activation of Hog1 is not sufficient for expression of copper-resistance. We propose that a defect in GPI anchor synthesis has multiple consequences, including activation of the Hog1 MAP kinase cascade and conferring copper resistance. PMID- 11922109 TI - Effects of yogurt supplemented with brewer's yeast cell wall on constipation and intestinal microflora in rats. AB - Brewer's yeast cell wall (BYC) is a valuable foodstuff that prevents constipation in rats. In this study, the effects of yogurt supplemented with brewer's yeast cell wall (BYC-Y) on constipated male Sprague-Dawley rats induced by loperamide (2 mg/kg body weight) were investigated. The preventive effect of BYC-Y on experimentally constipated rats was examined and compared with that of nonfiber diet (Control) and standard yogurt (Yogurt) as the control diets. The number of feces and fecal dry weight were greater in rats fed with BYC-Y than in those fed with Control or Yogurt. Moreover, the effect of BYC on the improvement of cecal microflora in constipated experimental model rats was evaluated. The number of Lactobacillus, Eubacterium, and total anaerobes per unit cecal contents increased (p<0.03) as a result of BYC ingestion. These findings indicate that BYC-Y or BYC is effective for the alleviation of constipation and bowel environment. PMID- 11922110 TI - Hypocholesterolemic effect of indigestible fraction of Chlorella regularis in cholesterol-fed rats. AB - The effects of Chlorella regularis powder (CP) and Chlorella regularis indigestible fraction (CIF) on serum and liver lipid concentrations and on fecal steroid excretion were estimated in rats fed diets containing 5 g/kg cholesterol and 2.5 g/kg sodium cholate. The ingestion of 12.7% CP or 5.3% CIF did not influence food intake or growth. CP and CIF decreased the levels of serum cholesterol, but had no effect on the levels of serum triacylglycerol and phospholipid. Liver cholesterol contents were lower in the CP and CIF groups than in the control group, but CP and CIF did not affect liver triacylglycerol content. CP and CIF increased the total amount of fecal neutral steroids excreted, but did not modify the total bile acid excretion. However, the soluble bile acid concentrations of reconstituted fecal water in the rats fed CP and CIF diets were lower than the control value. Moreover, CP and CIF had a high bile acid binding capacity in vitro. These results indicated that CIF had a hypocholesterolemic effect and enhanced fecal neutral steroid excretion while decreasing the soluble fecal bile acid concentration. PMID- 11922111 TI - Effects of carnitine coingested caffeine on carnitine metabolism and endurance capacity in athletes. AB - The purpose of this study was to examine whether caffeine (CAF), carnitine (CAR), or CAF+CAR mixture administration affects exercise endurance time via carnitine metabolism. Water (CON), CAF, CAR, or CAF+CAR mixture was administered to five male rugby athletes participating in this study by a randomized double-blind fashion who were made to ride a cycle ergometer for exercise. The CAF effect on exercise endurance time was small, but the CAR trial significantly increased the exercise endurance time compared with CON trial; a further CAF+CAR mixture trial had greater effects on the exercise endurance time than those of a CON, CAF, or CAR trial. A CAR or CAF+CAR mixed trial increased urinary nonesterified carnitine (NEC) and total carnitine (TCAR), but no changes were observed in acid-soluble acylcarnitine (ASAC) and acid-insoluble acylcarnitine (AIAC) excretion. A CAR or CAF+CAR mixed trial resulted in higher levels of plasma NEC, ASAC, and TCAR fractions than the CON and CAF trials did on exhaustion time. Total cholesterol, triglyceride, and free fatty acid in blood were significantly increased at exhaustion time, but they were not affected in the CAF or the CAR trial. These results suggest that carnitine ingestion could promote fat oxidation, resulting in higher endurance performance in athletes, and especially these ergogenic effects of carnitine coingested with caffeine may be greater than those of carnitine alone. PMID- 11922112 TI - Cooperation of divalent ions and thiamin diphosphate in regulation of the function of pig heart pyruvate dehydrogenase complex. AB - The role of Mg2+, Ca2+, and Mn2+ in regulation of purified pig heart pyruvate dehydrogenase complex (PDC) containing endogenous thiamin diphosphate (TDP) was studied. It was found that the effects of the cations depended on the presence of exogenous TDP. In the absence of added TDP, the divalent cations led to a shortening of a lag phase of the PDC reaction and a strong reduction of the Km value for pyruvate. The relative efficiency of the three types of ions are presented as follows: Mn2+>Ca2+>Mg2+. The other sources claim that in the presence of exogenous TDP, which alone strongly increased the affinity of PDC for pyruvate, any significant additional effects of the cations were not observed. However, Mg2+, Ca2+, and Mn2+ decreased the Km value for CoA in both cases, the absence and presence of exogenous TDP, in approximately a similar extent (about twofold). The affinity of PDC for NAD+ seems to be not sensitive to the presence of the divalent cations. The data obtained suggest that Mg2+, Ca2+, and Mn2+ can cooperate with TDP as positive regulatory effectors of pig heart PDC on the level of pyruvate dehydrogenase and lipoamide acetyltransferase components of the complex. PMID- 11922113 TI - Myocardial stereological adaptations in wistar rats fed with different high-fat diets during 18 months. AB - This study has the purpose of investigating the influence of different high-fat experimental diets on myocardial structure in rats. Twenty-seven male rats were fed from 21 d old (postnatal age) until 18 mo old with one of the following supplemented diets: soybean oil (S) (n= 6), canola oil (CA) (n= 8), or lard and egg yolk (LE) (n= 6) or canola oil+ lard and egg yolk (CA+LE) (n=7). The blood pressure (BP) was measured, and after the sacrifice the cardiac biometry and the myocardial stereology were determined: cross-sectional area of cardiomyocyte (A), volume density (Vv), surface density (Sv), and length density (Lv) in relation to the cardiomyocytes (cm), connective tissue (ct), and blood vessels (v). The CA group rats had lower BP, A[cm], and Vv[ct]; they had greater Vv[cm], Sv[cm], Vv[v], Lv[v], and Sv[v] than the other groups. The S rats had intermediary values for the myocardium and blood vessel parameters between the CA and LE group rats. These results support the notion that the long-term use of canola oil in the diet is better to preserve the myocardium structure, including microvascularization, than soybean oil or lard and egg yolk. PMID- 11922114 TI - Antifatigue effect of fresh royal jelly in mice. AB - We investigated the antifatigue effect of royal jelly (RJ), which had been stored at -20 degrees C from immediately after collection, in male Std ddY mice. The mice were accustomed to swimming in an adjustable-current swimming pool, then subjected to forced swimming five times during 2 wk, and the total swimming period until exhaustion was measured. They were separated into three groups with equal swimming capacity, which were administered RJ, RJ stored at 40 degrees C for 7 d (40-7d RJ), or the control solution including casein, cornstarch, and soybean oil before swimming. All mice were forced to swim for 15 min once; then the maximum swimming time to fatigue was measured after a rest period. The swimming endurance of the RJ group significantly increased compared with those of the other groups. The mice in the RJ group showed significantly decreased accumulation of serum lactate and serum ammonia and decreased depletion of muscle glycogen after swimming compared with the other groups, whereas there was no significant difference between the 40-7d RJ group and the control group in these parameters after swimming. A quantitative analysis of constituents in RJ showed that 5 7-kDa protein, which we previously identified as a possible freshness marker of RJ, was specifically degraded in RJ stored at 40 degrees C for 7 d, whereas the contents of various vitamins, 10-hydroxy-2-decenoic acid, and other fatty acids in RJ were unchanged. These findings suggest that RJ can ameliorate the physical fatigue after exercise, and this antifatigue effect of RJ in mice seems to be associated with the freshness of RJ, possibly with the content of 5 7 kDa protein. PMID- 11922115 TI - Analytical method of measuring tea catechins in human plasma by solid-phase extraction and HPLC with electrochemical detection. AB - We developed an analytical method for measuring tea catechins in plasma by solid phase extraction (SPE), followed by HPLC with a coulometric electrochemical detector. The plasma was mixed with an equal volume of acetonitrile to precipitate protein, and catechins in the resulting supernatant were extracted by SPE, using a C18 cartridge. To correct the extraction efficiency, ethyl gallate was simultaneously added with acetonitrile as an internal standard. Plasma samples were treated in microtubes, and evaporation and SPE were performed by the use of a vacuum centrifuge and vacuum manifold for SPE. The use of these instruments allowed the handling of a large number of samples simultaneously. In this method, (-)-epicatechin (EC), (-)-epicatechin-3-O-gallate (ECg), (-) epigallocatechin (EGC), (-)-epigallocatechin-3-O-gallate (EGCg), and ethyl gallate could be detected as a single peak with high sensitivity. For an analysis of the conjugated form of catechins, plasma samples were treated with glucuronidase and sulfatase. Type H-2 beta-glucuronidase effectively digested the conjugated forms, and the enzyme also converted EGCg and ECg to their nongallated form. When the concentrations of catechins in plasma were analyzed in subjects who took a single dose of catechin liquid, the concentration of free EGCg in plasma reached a maximum of 300 nM at 1 h after intake; those of the other free form of catechins increased only slightly after the intake. The concentration of total catechins (free+conjugated forms) in plasma increased up to 2 h after the intake. PMID- 11922116 TI - Stars to eyes by adaptive optics. PMID- 11922117 TI - Overestimation of heights in virtual reality is influenced more by perceived distal size than by the 2-D versus 3-D dimensionality of the display. AB - One important aspect of the pictorial representation of a scene is the depiction of object proportions. Yang, Dixon, and Proffitt (1999 Perception 28 445-467) recently reported that the magnitude of the vertical-horizontal illusion was greater for vertical extents presented in three-dimensional (3-D) environments compared to two-dimensional (2-D) displays. However, because all of the 3-D environments were large and all of the 2-D displays were small, the question remains whether the observed magnitude differences were due solely to the dimensionality of the displays (2-D versus 3-D) or to the perceived distal size of the extents (small versus large). We investigated this question by comparing observers' judgments of vertical relative to horizontal extents on a large but 2 D display compared to the large 3-D and the small 2-D displays used by Yang et al (1999). The results confirmed that the magnitude differences for vertical overestimation between display media are influenced more by the perceived distal object size rather than by the dimensionality of the display. PMID- 11922118 TI - A matching advantage for dynamic human faces. AB - In a series of three experiments, we used a sequential matching task to explore the impact of non-rigid facial motion on the perception of human faces. Dynamic prime images, in the form of short video sequences, facilitated matching responses relative to a single static prime image. This advantage was observed whenever the prime and target showed the same face but an identity match was required across expression (experiment 1) or view (experiment 2). No facilitation was observed for identical dynamic prime sequences when the matching dimension was shifted from identity to expression (experiment 3). We suggest that the observed dynamic advantage, the first reported for non-degraded facial images, arises because the matching task places more emphasis on visual working memory than typical face recognition tasks. More specifically, we believe that representational mechanisms optimised for the processing of motion and/or change over-time are established and maintained in working memory and that such 'dynamic representations' (Freyd, 1987 Psychological Review 94 427-438) capitalise on the increased information content of the dynamic primes to enhance performance. PMID- 11922119 TI - 'United' we stand. PMID- 11922120 TI - A double neon colour illusion. AB - A new visual illusion is presented. When two neon-coloured illusory bands overlap each other, the filling-in at the region of overlap is perceived as different from those of the overlapping bands, resulting in an additional illusory shape at the region of overlap. An experiment with ten naive participants was performed to measure the illusory percept. The results show that illusory surfaces may interact with each other, and, to a certain extent, create new illusory percepts. PMID- 11922121 TI - Context and the motion aftereffect: occlusion cues in the test pattern alter perceived direction. AB - A horizontally moving vertical grating viewed through a diamond-shaped aperture can be made to appear to move either upwards or downwards by introduction of appropriate depth-ordering cues at the boundaries of the aperture (Duncan et al. 2000 Journal of Neuroscience 20 5885-5897). The grating is perceived to move towards (and sliding under) occluding 'near' surfaces, and parallel to 'far' surfaces. Here we show that these depth-ordering cues affect the perceptual interpretation of the motion aftereffect (MAE) as well. After adaptation to unambiguous horizontal motion, the MAE direction deviates from horizontal towards near surfaces. However, the influence of depth-ordering cues on the illusory motion of the MAE is generally less than that seen for 'real' motion. Implications for theories of depth-motion and depth-MAE interactions are discussed. PMID- 11922122 TI - Scintillating lustre and brightness induced by radial lines. AB - Gray disks inserted into the central gaps of an Ehrenstein pattern appear to lighten up and scintillate with each movement of the eye or stimulus pattern. We call this phenomenon scintillating lustre. Both phenomena-illusory brightness and scintillating lustre-depend on the presence of the radial inducing lines converging onto the gaps. Without the radii the gray disks appear matte. Using parametric stimulus variation, we show that the strength of scintillating lustre covaries with line-induced brightness enhancement when the length, width, number, and contrast of the radial lines, as well as the size of the gaps in the Ehrenstein figure, are varied. Following the proposal by Anstis (2000, Vision Research 40 2551-2556), we suggest that lustre results from a competition between the ON and OFF visual pathways. Whereas Helmholtz's binocular gloss is elicited by stereoscopically fused incremental and decremental stimuli, the present study demonstrates that lustre can also arise from the interaction between line-induced brightness (illusory increment) and a dark gray disk (physical decrement). PMID- 11922123 TI - A planar and a volumetric test for stereoanomaly. AB - Stereoanomaly is the failure to see differences in depth when the viewer is presented with stimuli having different magnitudes of stereoscopic disparity. In the absence of eye movements, everyone suffers from stereoanomaly for extremely large disparities. Typically, such disparities are seen at the same depth as monocular stimuli. However, about 30%, of the population exhibit some form of stereoanomaly even for very small disparities, provided eye movements are avoided. In some cases, the sign of the disparity will be confused, and the perceived depth will be incorrectly seen as 'behind' rather than 'in front of' the fixation point, for example. Because anomalies provide useful information about perceptual mechanisms, tests that measure and quantify the extent of a blindness are important investigative tools for research. Here we offer two easy to-administer tests for stereoanomaly. The first test is based on depth judgments of two bars relative to a fixation point. The second test involves judgments of volumetric stimuli, seen stereoscopically. In each case, subjects indicate depth by setting a rectangle (with fixed base) to match the perceived depth. Although both tests are correlated, some differences in stereo processing are seen, depending upon whether or not the stimuli are presented near the point of fixation. PMID- 11922124 TI - Conditions under which stereopsis and motion perception are blind. AB - We describe modified random-dot stereograms in which the corresponding elements differ from non-corresponding elements in colour, size, and luminance. Despite these visible differences between the elements, depth perception collapses when the spatially integrated luminous flux is similar for the corresponding and non corresponding elements. Our results suggest that a low-pass spatial filter precedes the mechanism that recognises disparity. A similar phenomenon is observed for the perception of coherent motion in random-dot kinematograms. Our modified stereograms and kinematograms may find other uses when experimenters wish to study the contribution of colour to visual processes and require a method of eliminating edge artifacts. PMID- 11922125 TI - Is lightness induction a pictorial illusion? AB - Lightness induction, or simultaneous lightness contrast (we prefer the term lightness induction since contrast has another meaning in the visual literature, namely, the relative intensity of the stimulation), was studied for a 3-D object (Adelson's wall of blocks) and its 2-D pictorial representations. A statistically significant lightness induction effect was found only for the pictures but not for the 3-D object. No lightness induction effect was found for the 3-D object under either monocular or binocular viewing conditions. PMID- 11922126 TI - The pursuit of Leonardo's constraint. AB - Leonardo da Vinci (1452-1519) identified two stimulus situations that cannot be painted faithfully on a canvas: (a) when two objects are located in the same direction with respect to the painter's head, and (b) when parts of a surface are visible to one eye, but occluded from the other eye. He analysed these situations in terms of rays being emitted from the two eyes and, aside from the origin of the rays, the projective geometry he used was correct. His analyses showed that what can be seen from two vantage points cannot be represented on a canvas, because a 'correct' painting must be created from a single 'station point'. He was struck by the consequence of this fact that the depth seen on a canvas cannot match that of viewing the scene with two eyes. Subsequent visual scientists focused on Leonardo's observation about the lack of vivid depth in a picture. We argue that a complete understanding of what we see in the two stimulus situations requires consideration of visual direction in addition to visual depth. More specifically, we argue that the visual directions of the two objects, (a) above, and the visual direction of the monocular areas, (b) above, are dependent upon the constraint that two opaque objects cannot be represented in the same direction. Demonstrations that readers can perform, and that support this argument, are provided on the Perception website at http://www.perceptionweb.com/perc0102/ono.html. PMID- 11922127 TI - Color constancy, lightness constancy, and the articulation hypothesis. PMID- 11922128 TI - Articulation effects in lightness: historical background and theoretical implications. AB - The concept of articulation was first introduced by Katz [1935 The World of Colour (London: Kegan Paul, Trench, Trubner & Co)] to refer to the degree of complexity within a field. Katz, who created the basic research methods for studying lightness constancy, found that the greater the degree of articulation within a field of illumination, the greater the degree of constancy. Even though this concept has been largely forgotten, there is much empirical evidence for Katz's principle, and the effects on lightness are very strong. However, when articulation is increased within a framework that does not coincide with a region of illumination, constancy is weakened. Kardos (1934 Zeitschrift fur Psychologie Erganzungband 23) advanced the concept of co-determination, according to which the lightness of a surface is determined relative to more than one field of illumination. Gilchrist et al (1999 Psychological Review 106 795-834) argue that the fields concept should be replaced by the more operational frameworks concept and that a wide variety of lightness errors can be explained by a modification of the Katz principle: the greater the articulation within a perceptual framework, the stronger the anchoring of lightness values within that framework. PMID- 11922129 TI - Scene articulation: dependence of illuminant estimates on number of surfaces. AB - The ability of observers to detect changes in illuminant over two scenes containing different random samples of reflecting surfaces was determined in an experiment with Mondrian-like patterns containing different numbers of coloured patches. Performance was found to improve as the number of patches increased from 9 to 49. In principle, observers could have used space-average scene colour as the cue ('grey-world' hypothesis) or the colour of the brightest surface in the scene ('bright-is-white' hypothesis), as the two cues generally covary. In a second experiment, observers matched illuminants across different patterns in which the space-average cue and the brightest-patch cue were independently manipulated. The articulation of the patterns was varied: the number of patches increased from 49 (patch width 1 deg visual angle) to over 50000 (patch width 0.03 deg), while the gamut of colours was held constant. Space-average colour was found to be the dominant cue with all patterns except for those with the largest patches. PMID- 11922131 TI - Colour constancy in goldfish and man: influence of surround size and lightness. AB - Colour constancy was investigated by using a series of 10 simultaneously presented surface colours ranging in small steps from green through gray to red purple. Goldfish were trained to select one medium test field when the entire setup was illuminated with white light. In the tests, either red or green illumination was used. Colour constancy, as inferred from the choice behaviour, was perfect under green illumination when the test fields were presented on a gray or a white background, but imperfect on a black background. Under red illumination and a white background, however, colour constancy was overcompensated. Here, a colour contrast effect was observed. The influence of background lightness was also found when the surround was restricted to a narrow annulus of 4-11 mm width (test field diameter: 14 mm). By applying colour metrics it could be shown that the von Kries coefficient law can describe the overall effect of colour constancy. For an explanation of the effect of surround size and lightness, lateral inhibitory interactions have to be assumed in addition, which are also responsible for simultaneous colour contrast. Very similar results were obtained in experiments with the same colours in human subjects. They had to name the test field appearing 'neutral' under the different illumination and surround conditions, as tested in the goldfish experiment. PMID- 11922130 TI - Articulation: brightness, apparent illumination, and contrast ratios. AB - Luminance edges in the environment can be due to regions that differ in reflectance or in illumination. In three experiments, we varied the spatial organization of 10 achromatic (simulated) surfaces so that some arrangements were consistent with an ecologically valid and parsimonious interpretation of 5 surfaces under two different illuminants. A constant contrast-ratio along a luminance edge in the scene allows this interpretation. The brightness of patches in this condition was compared to their brightness with minimally different spatial arrangements that fail to maintain the constant contrast-ratio criterion. When the spatial arrangement of the 10 surfaces included a luminance edge satisfying the constant contrast-ratio criterion, brightness changed systematically, compared to arrangements without such a luminance edge. We account for the results by positing that a luminance edge with a constant contrast-ratio segments the scene into regions of lower and higher illumination, with the same effect as a difference in real physical illumination: all else equal, a given surface appears brighter under higher than under lower illumination. PMID- 11922132 TI - Articulation in the context of edge classification. AB - Many researchers believe the human visual system classifies luminance edges into those produced by reflectance edges or those produced by illumination edges, yet this classification process is not completely understood. One suggestion is that heuristics are used for edge classification. For example. specific contrast relationships at the luminance edge ('codirectional contrast invariance' and 'transversal luminance-ratio preserving') may distinguish an illumination edge from a reflectance edge on the one hand, and from a translucent edge on the other. Distinct from luminance junctions, these features are global characteristics of the luminance pattern that make distinguishing between different types of edge easier with more highly articulated scenes. I demonstrate that apparent translucency, nonreversing X-junctions, and single-reversing X junctions are insufficient on their own to produce the lightness illusion of Adelson's well-known tile pattern. While tolerating violations of the codirectional contrast invariance and transversal-luminance-ratio presersving without reversing the sign of contrast, the visual system is quite sensitive to such contrast reversal at the luminance edge. I account for this by suggesting that humans process lightness in terms of an ordinal, rather than interval, (or ratio) scale. PMID- 11922133 TI - Second-order statistics of colour codes modulate transformations that effectuate varying degrees of scene invariance and illumination invariance. AB - We argue, from an ethology-inspired perspective, that the internal concepts 'surface colours' and 'illumination colours' are part of the data format of two different representational primitives. Thus, the internal concept of 'colour' is not a unitary one but rather refers to two different types of 'data structure', each with its own proprietary types of parameters and relations. The relation of these representational structures is modulated by a class of parameterised transformations whose effects are mirrored in the idealised computational achievements of illumination invariance of colour codes, on the one hand, and scene invariance, on the other hand. Because the same characteristics of a light array reaching the eye can be physically produced in many different ways, the visual system, then, has to make an 'inference' whether a chromatic deviation of the space-averaged colour codes from the neutral point is due to a 'non-normal', ie chromatic, illumination or due to an imbalanced spectral reflectance composition. We provide evidence that the visual system uses second-order statistics of chromatic codes of a single view of a scene in order to modulate corresponding transformations. In our experiments we used centre surround configurations with inhomogeneous surrounds given by a random structure of overlapping circles, referred to as Seurat configurations. Each family of surrounds has a fixed space-average of colour codes, but differs with respect to the covariance matrix of colour codes of pixels that defines the chromatic variance along some chromatic axis and the covariance between luminance and chromatic channels. We found that dominant wavelengths of red-green equilibrium settings of the infield exhibited a stable and strong dependence on the chromatic variance of the surround. High variances resulted in a tendency towards 'scene invariance', low variances in a tendency towards 'illumination invariance' of the infield. PMID- 11922134 TI - The influence of chromatic and achromatic variability on chromatic induction and perceived colour. AB - Judgments of the colour of a surface are influenced by the colour of the surrounding. To determine whether only the average colour of the surrounding matters, or also the chromatic variability, judgments in colourful scenes are often compared with ones in which a target is surrounded by a plain background that provides the same average physical illumination of the retina as the colourful scene. The variability sometimes makes a difference (eg Shevell and Wei, 1998 Vision Research 38 1561-1566), and sometimes it does not (eg Brenner and Cornelissen, 1998 Vision Research 38 1789-1793). Is this because of the nonlinearity in cone responses? We designed scenes that stimulated the cones in an equivalent manner, both on average and in terms of variability, and yet differed markedly in chromatic variability. The more colourful surroundings had considerably less influence on subjects' colour judgments. We conclude that early cone-specific regulation of sensitivity cannot be responsible for the change in perceived colour, and deduce that chromatic induction takes place after contrast gain control. PMID- 11922135 TI - An empirical study of the traditional Mach card effect. AB - The traditional achromatic Mach card effect is an example of lightness inconstancy and a demonstration of how shape and lightness perception interact. We present a quantitative study of this phenomenon and explore the conditions under which it occurs. The results demonstrate that observers show lightness constancy only when sufficient information is available about the light-source position, and the perceptual task required of them is surface identification rather than direct colour-appearance matching. An analysis and comparison of these results with the chromatic Mach card effect (Bloj et al 1999 Nature 402 877 879) demonstrate that the luminance effects of mutual illumination do not account for the change in lightness perception in the traditional Mach card. PMID- 11922136 TI - Surface-illuminant ambiguity and color constancy: effects of scene complexity and depth cues. AB - Two experiments were conducted to study how scene complexity and cues to depth affect human color constancy. Specifically, two levels of scene complexity were compared. The low-complexity scene contained two walls with the same surface reflectance and a test patch which provided no information about the illuminant. In addition to the surfaces visible in the low-complexity scene, the high complexity scene contained two rectangular solid objects and 24 paper samples with diverse surface reflectances. Observers viewed illuminated objects in an experimental chamber and adjusted the test patch until it appeared achromatic. Achromatic settings made tinder two different illuminants were used to compute an index that quantified the degree of constancy. Two experiments were conducted: one in which observers viewed the stimuli directly, and one in which they viewed the scenes through an optical system that reduced cues to depth. In each experiment, constancy was assessed for two conditions. In the valid-cue condition, many cues provided valid information about the illuminant change. In the invalid-cue condition, some image cues provided invalid information. Four broad conclusions are drawn from the data: (a) constancy is generally better in the valid-cue condition than in the invalid-cue condition: (b) for the stimulus configuration used, increasing image complexity has little effect in the valid cue condition but leads to increased constancy in the invalid-cue condition; (c) for the stimulus configuration used, reducing cues to depth has little effect for either constancy condition: and (d) there is moderate individual variation in the degree of constancy exhibited, particularly in the degree to which the complexity manipulation affects performance. PMID- 11922137 TI - Neuroserpin, a neuroprotective factor in focal ischemic stroke. AB - Because recent studies have indicated that tissue plasminogen activator (tPA) aggravates neurodegenerative processes in many neural pathologies, we studied whether the endogenous tPA antagonist neuroserpin has a neuroprotective effect in an animal model of focal ischemic stroke. After induction of a focal ischemic stroke in the mouse by occlusion of the middle cerebral artery, we found that microglial cells accumulated in the marginal zone of the infarct are the most important source for both plasminogen activators, tPA and uPA. To investigate the effect of neuroserpin on the size and the histology of the infarct we produced transgenic mice overexpressing neuroserpin approximately sixfold in the nervous system. In the brain of these mice the total tPA activity in the uninjured tissue was strongly reduced. After induction of a focal ischemic stroke in the transgenic mice by a permanent occlusion of the middle cerebral artery (MCA), the infarcts were 30% smaller than in the wild-type mice. Immunohistochemical analyses and in situ hybridization revealed an attenuation of the microglial activation in the reactive zone. Concomitantly, the microglial production of tPA and uPA, as well as the PA-activity in the infarct region was markedly reduced. Thus, our results indicate that neuroserpin reduces microglial activation and, therefore, the PA activity and has a neuroprotective role after focal ischemic stroke. PMID- 11922138 TI - Notch signaling can inhibit Xath5 function in the neural plate and developing retina. AB - Neuronal differentiation is regulated by both positive and negative regulatory factors; however, precisely how these factors interact to regulate retinogenesis is still unclear. We have examined the ability of the Notch pathway to modulate the function of the basic helix-loop-helix factor Xath5. Overexpression of Xath5 by RNA injection into cleavage-stage blastomeres promotes ectopic neurogenesis at neural plate stages and ganglion cell differentiation in the developing retina. We found that these activities of Xath5 could be inhibited by coexpression of activated Notch. Notch inhibition of Xath5 function was reversed by coexpression with the zinc finger protein X-MyT1. The Notch effector enhancer-of-split related 1 (ESR1) also blocked Xath5 activity but efficient inhibition by ESR1 required the DNA binding basic domain and the conserved WRPW motif. In addition, ESR1 inhibited the ability of Xath5 to directly activate the expression of XBrn3d, a transcription factor involved in retinal ganglion cell development. Xath5 could upregulate expression of X-Delta-1, ESR1, and ESR3, suggesting that Xath5 participates in a regulatory loop with the Notch pathway. PMID- 11922139 TI - Association between integrin-dependent migration capacity of neural stem cells in vitro and anatomical repair following transplantation. AB - In previous transplantation studies using neural stem cell lines immortalized by the temperature-sensitive SV40 large T-antigen, we have shown that animals with experimental hippocampal lesions resulting from four vessel occlusion recover spatial memory functions more effectively when grafted with the MHP36 cell line than with the MHP15 cell line [Gray et al. (1999). Philos. Trans. R. Soc. London Biol. Sci. 354:1407-1421]. In the present study, we have investigated the cellular and molecular basis of these differences in repair capacity both in vivo and in vitro. Using the same model of hippocampal damage we have shown that following transplantation MHP36 cells migrate and align within the damaged CA1 of the ipsilateral hippocampus. MHP15 cells, in contrast, migrate in a more indiscriminate pattern that does not reflect the anatomy of the region. To analyze the migratory properties of these two cell lines in more detail, we performed migration assays at a nonpermissive temperature on the extracellular matrix substrates laminin, fibronectin, and vitronectin. These showed that MHP36 cells have a greater migration potential than the MHP15 cells. While the pattern of cell surface extracellular matrix receptors of the integrin family was identical in both cell lines, the different degrees of migration on vitronectin were both blocked by inhibitors of alphaV integrins. Differences in integrin signaling therefore contribute to the greater migration potential of the repairing MHP36 cell line. PMID- 11922140 TI - Emx2 promotes symmetric cell divisions and a multipotential fate in precursors from the cerebral cortex. AB - Distinct sets of precursor cells generate the mammalian cerebral cortex. During neurogenesis most precursors are specified to generate a single cell type and only few are multipotent. The cell-intrinsic molecular determinants of these distinct lineages are not known. Here we describe that retroviral transduction of the transcription factor Emx2 in precursors from the cerebral cortex results in a significant increase of large clones that are generated mostly by symmetric cell divisions and contain multiple cell types, comprising neurons and glial cells. Thus, Emx2 is the first cell-intrinsic determinant able to instruct CNS precursors towards a multipotential fate. To evaluate the role of endogenous Emx2 in cortical precursors, we examined cell division in Emx2-/- mice. These analyses further supported the role of endogenous Emx2 in the regulation of symmetric cell divisions in the developing cortex. PMID- 11922141 TI - A short cytoplasmic domain of the amyloid precursor protein induces apoptosis in vitro and in vivo. AB - The amyloid precursor protein presents several cleavage sites leading to the release of its entire C-terminal domain into the cytoplasm. During apoptosis, this C-terminal domain can be cleaved at amino acid 664 by caspases 3, 6, and 8 and can thus generate two peptides N- and C-terminal to amino acid 664 (C31). Recently, it was shown that the C31 induces apoptosis after transfection into N2A and 293 T cell lines. We have analyzed here, by internalization into neurons, the physiological consequences of the entire C-terminal domain (APP-Cter) and of its membrane proximal sequence corresponding to the N-terminal peptide unmasked after caspase cleavage. We find that whereas micromolar concentrations of APP-Cter are harmless, the peptide extending from the membrane (amino acid 649) to the caspase cleavage site (amino acid 664) in the same range of concentrations induces DNA fragmentation, cleavage of actin at a caspase-sensitive site, and activates caspase 3. A mutated version of this sequence (tyrosine 653 replaced by an aspartate) abolishes the effect in vitro and in vivo. Taken together, this report suggests the existence of a new mechanism contributing to Alzheimer's Disease associated cell death. PMID- 11922142 TI - M1 muscarinic acetylcholine receptors activate extracellular signal-regulated kinase in CA1 pyramidal neurons in mouse hippocampal slices. AB - Activation of extracellular signal-regulated kinases (ERK) is crucial for many neural functions, including learning, memory, and synaptic plasticity. As muscarinic acetylcholine receptors (mAChR) modulate many of the same higher brain functions as ERK, we examined mAChR-mediated ERK activation in mouse hippocampal slices. The cholinergic agonist carbachol caused an atropine-sensitive ERK activation in the dendrites and somata CA1 pyramidal neurons. To determine the responsible mAChR subtype, we combined pharmacologic and genetic approaches. Pretreatment with M1 antagonists inhibited ERK activation. Furthermore, mAChR induced ERK activation was absent in slices from M1 knockout mice. ERK activation was normal in slices derived from other mAChR subtype knockouts (M2, M3, and M4), although these other subtypes are expressed in many of the same neurons. Thus, we demonstrate divergent functions for the different mAChR subtypes. We conclude that M1 is responsible for mAChR-mediated ERK activation, providing a mechanism by which M1 may modulate learning and memory. PMID- 11922143 TI - The Lnx family proteins function as molecular scaffolds for Numb family proteins. AB - Drosophila Numb functions as a cell fate determinant during neurogenesis. We isolated a novel mammalian protein, Lnx2, which interacts with mammalian Numb and Numblike. Lnx2 and the related Lnx1 are multimodular proteins that bind to Numb via their NPXY motifs. In addition, Lnx proteins form oligomers either via their PDZ domains binding to PDZ-binding consensus motifs located in their C-termini or by homophilic oligomerization of their RING fingers. Therefore, Lnx proteins may form large networks by homomeric binding. In situ hybridization analysis revealed complementary patterns of Lnx1 and Lnx2 expression in developing and adult brain, although in several structures they are present in the same cell populations. Moreover, their expression patterns overlap with those of the Numb proteins. Oligomerization of Lnx2 and Numb binding occurs simultaneously. Therefore, our findings suggest that Lnx proteins may serve as molecular scaffolds that localize unrelated, interacting proteins, such as Numb, to specific subcellular sites. PMID- 11922144 TI - Characterization of a new brain-derived proteoglycan inhibiting retinal ganglion cell axon outgrowth. AB - A proteoglycan was identified and isolated from physiological saline extracts of chick embryo brains by using a new monoclonal antibody (hybridoma clone mab Te38). The purified proteoglycan displayed an apparent molecular mass of 2500 3500 kDa, which became reduced to 370 and 600 kDa after digestion with chondroitinase ABC or chondroitinase AC. After additional treatment with keratanase the 600-kDa band was no longer detectable in Western blots. The specific epitope recognized by mab Te38 is an O-linked carbohydrate associated with the core protein. Tenascin-C, an extracellular matrix protein known to associate with several proteoglycans, copurified with the mab Te38 proteoglycan on the immunoaffinity column. Mab Te38 binds to the surface of nonneuronal cells; in sections from the primary visual system, expression is restricted to cells in the optic fissure, the dorsal optic nerve, and the chiasm. No retinal cells were found to express the mab Te38 epitope. The isolated molecule inhibited axon outgrowth from retinal explants when offered bound to a substrate consisting of either matrigel or collagen, chondroitinase treatment did not alter the inhibitory properties. The distribution and in vitro function of the Te38 proteoglycan indicate that it may serve a role in guidance of retinal ganglion cell axons. PMID- 11922146 TI - Differential modulation of sodium channel gating and persistent sodium currents by the beta1, beta2, and beta3 subunits. AB - Brain sodium channels are complexes of a pore-forming alpha subunit with auxiliary beta subunits, which are transmembrane proteins that modulate alpha subunit function. The newly cloned beta3 subunit is shown to be expressed broadly in neurons in the central and peripheral nervous systems, but not in glia and most nonneuronal cells. Beta1, beta2, and beta3 subunits are coexpressed in many neuronal cell types, but are differentially expressed in ventromedial nucleus of the thalamus, brain stem nuclei, cerebellar Purkinje cells, and dorsal root ganglion cells. Coexpression of beta1, beta2, and beta3 subunits with Na(v)1.2a alpha subunits in the tsA-201 subclone of HEK293 cells shifts sodium channel activation and inactivation to more positive membrane potentials. However, beta3 is unique in causing increased persistent sodium currents. Because persistent sodium currents are thought to amplify summation of synaptic inputs, expression of this subunit would increase the excitability of specific groups of neurons to all of their inputs. PMID- 11922145 TI - Fibroblast growth factor-2 inhibits myelin production by oligodendrocytes in vivo. AB - Fibroblast growth factor-2 (FGF-2) controls in part the timely differentiation of oligodendrocytes into the myelin-producing cells of the CNS. However, although differentiated oligodendrocytes express FGF receptors (R), the effect of FGF-2 on myelin-producing oligodendrocytes in vivo was unknown. In the present study, we show that delivery of FGF-2 into the cerebrospinal fluid of anaesthetized rat pups, aged postnatal day (P) 6 to 9, induced a severe loss of myelin in the caudal anterior medullary velum (AMV). Furthermore, we show that the caudal AMV was myelinated at the time of treatment, so the effects of FGF-2 represent a loss of myelin and not delayed differentiation. This was confirmed by injection of platelet-derived growth factor-AA (PDGF-AA), a factor known to affect the differentiation of PDGF-alphaR expressing oligodendrocyte progenitors, but which did not induce myelin loss in the caudal AMV and did not affect differentiated oligodendrocytes, which do not express PDGF-alphaR. Compared to controls treated with saline or PDGF-AA, FGF-2 induced an accumulation of PLP protein and MBP mRNA within the somata of myelin-producing oligodendrocytes. The results indicate that FGF receptor signalling disrupts myelin production in differentiated oligodendrocytes in vivo and interrupted the transport of myelin-related gene products from the oligodendrocyte cell body to their myelin sheaths. PMID- 11922147 TI - Medical treatment of ectopic pregnancy using parental methotrexate. AB - Between January and December 1999, we were able to diagnose five cases of unruptured ectopic pregnancy, using a transabdominal convex sector transducer. All the five patients presented with lower abdominal pains but were in stable clinical state. They were treated with two doses of 50mg methotrexate intravenously, 48 hours apart. Four of the patients had clinical, immunological, and sonographic evidence of resolution. The only failure was in the patient whose gestation sac was 4.8cm in its maximal diameter. She eventually had laparotomy and salpingectomy. PMID- 11922148 TI - Total antioxidant status in non-insulin-dependent diabetes mellitus patients in Ghana. AB - Total antioxidant status (TAS) was measured in thirty-five (35) non-insulin dependent diabetes mellitus (NIDDM) patients aged 40 to 65 (mean +/- SE 49.6 +/- 1.0) years. Patients were on diet and oral hypoglycaemic drug (Daonil) therapy with fasting plasma glucose (FPG) >7.8mmol/l. Similar measurements were carried out in thirty-four apparently healthy individuals within the same age range (mean + SE 46.3 +/- 1.1 years) and FPG <6.4 mmol/L. FPG was measured by glucose oxidase method and TAS by colorimetric method. Comparing the two groups, TAS was significantly reduced in the NIDDM patients (p<0.001). An inverse correlation between FPG and TAS suggests the existence of lower antioxidant defence in uncontrolled NIDDM. A good control of FPG accompanied with antioxidant therapy could help reduce free radical activity and minimise complications associated with increased free radical activity in diabetic patients. PMID- 11922149 TI - Clinical profile of acute otitis media among Nigerian children. AB - A one-year prospective clinical study to assess principal clinical symptoms, signs and bacteriological isolates in Nigerian children with acute otitis media. This is a multicentre study conducted from January-December, 1998 in three University Teaching Hospitals. A strict recruitment protocol was followed. Patient symptoms had to be less than one week before enrollment. Eighty-one children were recruited into the study. The major findings indicative of acute otitis media were ear pain (92.6%), fever (88.9%), redness of the tympanic membrane (89.9%) and loss of light reflex (84%). Rupture of the tympanic membrane occurred in 15 patients (18.5%) Staphylococcus aureus was the most common pathogen (50%) isolated from the ear swabs. A high index of suspicion is necessary to diagnose acute otitis media early in children especially those who may have fever as the main symptom. This will minimise the frequency of tympanic membrane perforations. PMID- 11922150 TI - Social characteristics, HIV/AIDS knowledge, preventive practices and risk factors elicitation among prisoners in Lagos, Nigeria. AB - BACKGROUND: Although many behavioral research studies and public enlightenment campaigns have been undertaken by both government and non-governmental organizations in the general public, no such study has been documented on prison inmates in Nigeria. This study aimed at documenting the social characteristics, HIV/AIDS knowledge and preventive practices of selected prisoners in Nigeria. It also elicited risk factors HIV/AIDS transmission in Nigeria prisons. METHODS: A cross-sectional study of prison inmates using an anonymous risk-factors identification questionnaire was undertaken in January 1997. The Kiri-kiri (maximum, medium and female) prisons were selected by balloting. Thereafter, two hundred and fifty two inmates were selected by systematic random sampling method using the full listing of all inmates as at the time of the survey. The study comprised of an interview session using a well structured questionnaire to seek information about their social data, their knowledge about HIV/AIDS including its transmission and preventive social data, and their indulgence in HIV/AIDS risky behaviour. RESULTS: The majority (53.6%) of the respondents were in the age group 20-29 years, 18 (7.1%) were less than 20 years old one of whom was in the maximum security prison and three were females (table 1). The majority (52%) had secondary education while 9.9% had tertiary education and 7.1% had no formal education. About 97.2% of the study population had heard about AIDS although only 20.6% had known or seen someone with AIDS before and about 34.1% knew the causative agent of AIDS. 60.3% knew the correct mode of transmission of AIDS. 15.5% claimed fidelity and 12.7 % claimed use of condom for casual sexual contact, were measures that could help prevent AIDS but 7.9% did not know any preventive measure. Since hearing about AIDS, 59.5% claimed to have taken steps to protect themselves. 42.7% of the 89 who had not taken any protective steps against AIDS had no knowledge of how to protect themselves. About 56.3% claimed to have used condom before although only 38.7% used it for their last sexual exposure while 28.2% claimed they used it for all casual sexual intercourse. Many (42.8%) said they knew that homosexuality was the most prevalent sexual practice in the prison while 28.6% claimed there was no sexual practice and 13.1% feigned ignorance of any sexual practices in the prisons. Many (53.2%) claimed to have multiple sexual partners although 94.8% denied any sexual practice whilst still in prison. CONCLUSION: This study demonstrated that (i) almost all THE prisoners studied had heard of AIDS although only a few had seen or known a case of AIDS; (ii) despite the fact that many of them knew the correct modes of transmission, many indulged in high risk behaviours for AIDS transmission; (iii) there is a considerable proportion of receptive naive inmates who stand the risk of being infected due to their high level of ignorance about HIV/AIDS. Well designed information, education and communication (IEC) programmes on AIDS with such formidable support structures as the provision of harm-reduction devices and risk reduction counselling are urgently recommended for the Nigerian prisoners to effectively combat the imminent HIV/AIDS epidemic among the prison inmates. PMID- 11922151 TI - Aetiology of endomyocardial fibrosis (EMF). AB - On epidemiological basis EMF behaves like a vector transmitted disease. The cardiac pathologies of EMF and HES are identical. In some cases of HES, hypereosinophilia may return to normal, leaving residual heart disease that is exactly like EMF. Most temporary residents from Europe and North America who developed EMF while resident in the endemic areas of Africa had hypereosinophilia that was induced by helminths. In our case studies from the EMF endemic areas of Nigeria, most children with acute idiopathic myocarditis associated with helminth induced hypereosinophilia, developed clinical EMF on follow up. We showed also that the rate of decline in the incidence of hypereosinophilia in EMF cases was significantly related to the duration of symptoms. Our studies and other observations show that EMF, like HES is a multiple system disease with similar organ damage. The morphologic evolution of cardiac damage in EMF appears similar to that reported for HES; with a stage of myocarditis/pericarditis, followed by a stage of cardiac necrosis, a stage of thrombosis and by the chronic fibrotic stage. Also during larval migration, all the helminths associated with EMF induce the same spectrum of damage in the central and peripheral nervous system, in the lungs, kidneys and skin, as are reported for HES. The cardiovascular damage reported for these worms (which include hypersensitivity vasculitis, acute myocarditis/ pericarditis) are also similar to what is reported for HES. Acute endomyocardial necrosis and thrombosis that are similar to what is found in HES, have been documented in Trichinella Spiralis and in filariasis. Increased cerium concentrations have been documented in the endocardium of EMF cases from South India. It remains to be established whether cerium excess, which is known to stimulate collagen synthesis does accelerate the process of endomyocardial fibrosis, following cardiac necrosis (which may have been triggered by helminths and the associated hypereosinophilia). PMID- 11922152 TI - Chronic hepatomegaly in steady state haemoglobin S children: some clinical and abdominal duplex ultrasonographic observations. AB - Chronic hepatomegaly (CH) is an index of severe disease in children with sickle cell anaemia (HbS). We studied 14HbS children with CH and two groups of controls made up of age and sex-matched HbA and HbS children without CH. The objective was to determine and compare hepatic perfusions in the children and also determine which clinical and ultrasonographic features are associated with CH. Hepatic perfusion was assessed using abdominal duplex ultrasonography which made possible the measurement of portal vein diameter (PVD) and the velocity of portal blood flow. The mean weights, blood pressure profiles and ultrasonographic splenic span and common bile duct diameters (BDD) did not differ significantly among the three groups. Similarly, the man PVDs of the three groups were not significantly different despite the large livers of the CH group. The CH group however showed linear growth faltering, a significantly lower mean haematocrit, faster pulse rate but paradoxically reduced hepatic portal blood flow velocity than HbA controls. Because of the disproportion of PVD relative to liver size in HbS patients with CH and the reduced portal blood flow velocity, hepatic perfusion is probably poorer in these patients. HbS children with CH therefore need careful management of hepatic and anaemic crises to prevent severe hypoxic liver damage. PMID- 11922153 TI - Lack of association between some risk factors and hepatitis B surface antigenaemia in children with sickle cell anaemia. AB - The risk factors for transmission of Hepatitis B virus (HBV) as well as the relationship between HBs Antigeneamia and some socio-cultural factors including socio-economic status of the families of the children, tribe, religion, rural or urban domicile and the number of persons per bedroom, were studied among 507 children with sickle cell anaemia (SCA) and 501 controls at the Jos University Teaching Hospital (JUTH). Twenty-three percent (22.7%) of the children with SCA were HBsAg positive compared with 19.6% among the control group. None of the risk factors studied, including blood transfusion, parenteral injections, intravenous infusions, hospitalization, ulcers, traditional uvulectomy, surgery, contact with case of known hepatitis, mass immunizations, circumcision, ear piercing and traditional scarifications was significantly associated with HBV infection in both children with SCA and control group and also between children with SCA compared with controls. There was also no association between HBs Antigenaemia and the number of blood transfusions, intravenous infusion or hospitalization in both children with SCA and controls. Similarly, there was no association between the socio-cultural factors studied and HBs Antigenaemia. The result of this study suggests that HBV infection is common in children in this environment and other factors yet to, be identified may play a very important role in the transmission of HBV infection. The high prevalence of HBsAg obtained in both children with SCA and controls, and the lack of identifiable association between the risk factos studied and HBsAntigenaemia suggest that these children may have equally been exposed earlier in life. PMID- 11922154 TI - Pain assessment in Nigerians--visual analogue scale and verbal rating scale compared. AB - The usefulness of 2 methods of pain assessment was determined in a cohort of Nigerians who had pain as a symptom and were receiving physiotherapy for various indication. The English and Yoruba versions of two Pain Rating Scales, the Visual Analogue Scale (VAS) and Verbal Rating Scale (VRS) with 1 to 4 or 1 to 5 intensity scales (VRS-4 or VRS-5) were employed for the assessment of pain in 100 patients. The mean Pain Score on the 4-point VRS Scale was 2.49 +/- 0.72 for the 5-point VRS 2.1 +/- 1.18 and for the VAS 4.93 +/- 2.5. Correlation analysis for corresponding groups of patients showed a significant positive relationship between the VAS and VRS-4 (r = 0.68 P<0.001) VAS and VRS-5 (r = 0.64 P <0.001) indicating that both VAS and VRS constitute useful tools for pain assessment in Nigerian patients. PMID- 11922155 TI - Evaluation of 56 trabeculectomy operations at Ago-Iwoye, Ogun State, Nigeria. AB - Records of 56 eyes of 34 patients who had trabeculectomy between February 1998 and October 1999 at the St. Mary's Catholic Eye Hospital Ago-Iwoye, Ogun State were reviewed retrospectively. There were 22 (64.7%) males and 12 (35.3%) females. Age range was from 18 years to 80 years, mean age 54 years. 22 patients had bilateral trabeculectomy and 12 patients had uniocular surgery. Intra ocular pressure control less than 21 mmHg post operatively was 74.1%; 96.2% was achieved with additional use of antiglaucoma medication; 3.7% could not be controlled on medication post operatively. Here was a statistically significant decrease in visual acuity post operatively. The Intra-operative and post-operative complications were minimal. PMID- 11922156 TI - Intestinal malrotation: experience in Zaria, Nigeria. AB - This report retrospectively reviews the presentation, management and outcome in 14 patients with intestinal malrotation. There were 7 males and 7 females, with a median age of 4 months (range 5 days-22 years). Most neonates and infants presented with persistent or recurrent vomiting while older children had recurrent abdominal pain. Five patients (36%) including 2, < 1 year and 3 older children developed midgut volvulus. Malrotation was an incidental finding at laparotomy for unrelated conditions in 2 patients (14%). Overall, preoperative diagnosis was not made in any patient. Associated congenital anomalies were present in 3 (21%) patients. Treatment was by Ladd's procedure in 12 (86%) patients and caecopexy in 2 (14%) without recurrence of symptoms in any. One patient each developed wound infection and adhesive intestinal obstruction requiring relaparotomy respectively. Mortality was 2 (14%) from overwhelming infection and hypokalaemia following midgut volvulus. Children presenting with recurrent or persistent abdominal pain, vomiting or failure to thrive may well have malrotation and should be investigated promptly and the condition corrected to avoid midgut volvulus which is attended by significant morbidity and mortality. PMID- 11922157 TI - Anterior abdominal wall reconstruction with fascia lata. AB - Anterior abdominal wall is a complex fasciomuscular structure, defects of which may arise as a result of infection, trauma, malignancy and herniation. Different techniques have been devised to repair these defects with varying result, availability and cost implications. In the communication, the use of fascia lata for repair of major anterior abdominal wall defect in five patients is reported. The result suggests that this is a useful technique that is associated with satisfactory outcome and minimal morbidity. PMID- 11922158 TI - Gastric outlet obstruction in Ogbomoso, Nigeria. AB - Forty-three patients were operated upon for gastric outlet obstruction over an eight year period. Six (14%) of these patients bad malignant gastric outlet obstruction. Three groups of procedures were used: truncal vagotomy and drainage (TV-D) in 32 patients, highly selective vagotomy and drainage (HSV-I)) in 8 patients, and gastric resection with or without vagotomy in 3 patients. Post operatively, it took an average of 6.6 days for patients to recommence oral intake (6.2 days for patients who underwent truncal vagotomy and 8.5 days for those who underwent highly selective vagotomy). There were no deaths; immediate post-operative morbidity included gastric atony in 21% and wound infection in 7% of the patients. Long-term complications include postvagotomy diarrhea in one patient and dumping in two patients. One patient had a recurrence of gastric outlet obstruction necessitating re-operation. PMID- 11922159 TI - Measles HI-antibody levels in Lagos children, Nigeria: a follow-up study to resurgence of measles in Lagos metropolis. AB - A total of 138 children aged between 0-36 months were bled and screened for measles haemagglutination inhibiting (HI)-antibody. Twenty seven children were from age group (0-4) months with 16 (59.3%) seropositive while, 67 and 44 were bled in the group (5-9) and > or = 12 months with 29 (43.3%) and 31 (70.5%) sero positive sera respectively. The distribution of the antibody titre pattern among various age groups suggest vaccination against measles should be at age 9 month. The antibody titres pattern of seroconverted vaccinees also showed that the vaccine administered at our study centre were potent enough to elicit good and protective immune response in vaccinees when given at age 9 month. Our study agrees with the current policy of vaccinating children against measles at age 9 months in Nigeria and should therefore be continued. Based on observations in this study, we suggest; continued administration of the Edmonston-Zagreb strain of measle vaccine currently-in-use in Nigeria, mother's breast-feeding of children for a longer period before the apporpriate age for vaccination, and the possible administration of Vitamin A-a potent immune enhancer, as a supplement during immunization programmes in Nigeria. The introduction of a booster dose of measles vaccine at elementary school age in order to care for the residual unvaccinated children may be a good strategy for measles eradication necessary to be considered in Nigeria and might probably forestall instances of measles outbreaks in children during their second year in school. PMID- 11922160 TI - Vitamin A status of pre-school children in Ibadan (South West Nigeria), risk factors and comparison of methods of diagnosis. AB - Vitamin A deficiency (VAD) and protein energy malnutrition (PEM), sharing common aetiological factors, are important public health problems in many developing, countries. A cross-sectional survey of the vitamin A status of 128 well nourished and 230 malnourished pre-school children was carried out in order to define factors associated with increased risks of VAD and also to determine the predictive values of CIC-T in identifying serum retinol of < 10 microg/dl in these children. The proportional morbidity rates of VAD defined by serum retinol concentrations (7.3%) and CIC-T (6.2%) was similar (p>0.05), and children aged < 3 years accounted for 70% of VAD cases. VAD occurred in 6.3% and 7.8% of well nourished and malnourished children respectively. The risk of VAD was increased following measles, history of persistent diarrhoea and wasting. The predictive value of CIC-T is highly dependent on CIC-T such that abnormal and normal smears classification appears to be very robust and predictive of serum retinol of < 10 microg/dl, with sensitivity of 83.3% (95%CI: 61.8-94.5), and specificity of 73.3% (95%CI: 68.3-78.5). Judging by the proportional morbidity rate in this study, VAD appears to be a significant public health problem in both malnourished and well nourished Nigerian children, especially in children < 3 years of age. The history of measles and persistent diarrhoea appear to increase the risk of VAD. The simplicity, sensitivity and specificity of CIC-T suggest that this procedure is a good screening tool for epidemiological survey of vitamin A status. PMID- 11922162 TI - Medical examination findings among army recruits in Ilorin. AB - This paper illustrates the need for proper medical examination of subjects or candidates registering or entering into new vocations particularly in the armed and paramilitary forces. Sixty army recruits who presented for medical examination at the University of Ilorin Teaching Hospital were studied. The study showed that nineteen (31.6%) of the subjects had positive pathological findings. These include: High blood pressure (20%) Visual refractive errors (5%) Remarkable chest X-ray changes (5%) Inguinal hernia (1.6%). It was concluded that candidates and subjects who wish to embark on new vocations would benefit from proper pre employment medical assessment and recommendations were made in that line. PMID- 11922161 TI - Prevalence of children with disabilities in central region, Ghana. AB - A cross sectional survey was conducted in Central Region, Ghana to determine the prevalence of children with disabilities. Forty-seven (1.8%) out of a total of 2,556 children under fifteen years of age had disabilities. About a third, had difficulty with movement of which post poliomyelitis infection disability formed the majority. Twelve children (25.5%) had difficulty with hearing and speech (deaf and dumb). Three children were reported as having epilepsy and 2 others had mental retardation. There was no case of total blindness, however, two children complained of poor vision and three others had a squint. Other disabilities identified, included one child each with kyphosis (hunch back), hydrocephalus (very large head) and extensive facial scars from burns. Disability was more common among children who had not been immunized or who did not have immunization card at the time of interview. The prevalence of disability was higher among female children in the rural community and in children with no formal education. Prevalence of disability increased with age. The prevalence of disability was 14.4 per 1,000 for children (1-5) years, 16.6 per 1000 for children (6-9) years and 3.7 per 1,000 for (10-15) years age group. Younger children had better immunization status and lower disability rates. Thirty percent of the children with disability said they were experiencing discrimination especially among the female children. This study has demonstrated that there is a need to intensify the preventive efforts at reducing the problem of childhood disability and to increase public support to the disabled children by avoiding discrimination and encouraging education and training of the disabled children. PMID- 11922163 TI - Pattern of suicides in Ile-Ife, Nigeria. AB - This is a study of pattern of autopsy findings in cases of completed suicides, with respect to age, sex and methods employed over a period of 11 years and 2 months. Suicides are generally reported to be rare in the less developed countries. Yet sudden deaths of indetermined aetiology are a well-recognized entity. Besides, it has been established that depressive disorders, a major cause of suicides, hitherto thought to be rare in Africa, is now recognized to be fairly common, presenting in a rather subtle form with features of somatization. Suicides in Ife were found to be 0.4 per 100,000 population with a higher incidence in males with a ratio 3.6 to 1. The majority of the victims were in the third decade of life. The majority of suicides were committed with Gammalin 20 and the local Dane gun. Those who used the gun applied it to the head and neck region in 76% of the cases. PMID- 11922164 TI - Serum electrolyte, urea and creatinine in clinically stable Nigerian newborn. AB - Serum electrolytes urea and creatinine of 80 clinically stable normal newborn comprising 60 preterm and 20 term appropriate for gestational age babies were prospectively studied. A negative correlation between the serum sodium, potassium, urea, creatinine and the gestational age was found. A statistical significant difference in the mean values of serum urea and creatinine in both preterm and term babies was obtained but there was no statistical significant difference in the mean value of serum sodium and potassium. PMID- 11922165 TI - Indigenous hospital technology in Nigeria: problems and recommendations. AB - In Nigeria, after gaining independence there arose a national desire for industrialization in order to reduce the high cost of sustaining national development. This desire has its accompanying problems which are examined with reference to indigenous hospital technology. Solutions to these problems are also suggested. PMID- 11922166 TI - Fibular on-lay graft in the management of radial giant cell tumour--a case report. AB - The management of bone loss following tumour resection poses a problem particularly in the upper limb where limb preservation is paramount. For bone loss less than 6cm, nonvascularized fibular graft has been advocated whereas in bone defects larger than this, vascularized fibular graft is the preferred option. In this case study, we have used a nonvascularized fibular on-lay graft (supplemented with cancellous bone graft) in the management of a distal radial bone loss of ten centimeters following resection of a giant cell tumour with remarkable success. PMID- 11922167 TI - Quadriceps tendon rupture in an adolescent. AB - Quadriceps tendon rupture is said to be rare in patients younger than 40 years of age. We report a case of quadriceps tendon rupture in a 13 year old athletic boy. PMID- 11922168 TI - Case report: difficult spinal anaesthesia for caesarian section in two obese pregnant patients. AB - Two obese patients presented for Caesarean Section for poor progress in labour. Spinal anaesthesia was chosen because of obesity, gravidity and worry about possible related airway problems in both patients. In both cases the needle seemed to be short. In case 1, no cerebrospinal fluid (CSF) was obtained, and she was subsequently done under General Anaesthesia (GA); in case 2, CSF was obtained when the needle was pressed firmly into the patients back but flow stopped if the pressure was released. These case reports serve to illustrate that although spinal anaesthesia for obese pregnant patients may seem the safer alternative, mechanical difficulty may be considerable and eventually a GA may be considered in spite of possible risk of difficult intubation. PMID- 11922169 TI - Nasopharyngeal carcinoma with distant metastasis. AB - We report a case of nasopharyngeal carcinoma with Chest Metastasis in a 38-year old man. The patient presented with nasal obstruction, epistaxis, a huge neck mass and conductive hearing loss in the right ear. Examination under anaesthesia revealed a mass in the nasopharynx, which was confirmed on histology to be squamous carcinoma. He responded remarkably well to external radiotherapy with disappearance of primary tumor and neck metastasis. One year later he presented with thoraco-lumbar spine pain and cough. The nasopharynx and neck remained free of tumor while radiographs demonstrated multiple metastasis to the lungs and vertebrae. PMID- 11922170 TI - Congenital peritoneal encapsulation of small intestine: a case report. AB - Congenital peritonel encapsulation of the small intestine is a rare malformation. It is associated with clinical features which may cause diagnostic difficulties and more often it is recognised only at exploratory laparotomy. We present a case in a 15 year old female, who was treated and she had a good result. Embryological explanation for the pathology is also considered. It is a disease entity which should be recognised, especially when encountered at laparotomy and treated appropriately. PMID- 11922171 TI - Going molecular. PMID- 11922172 TI - Focus on animal welfare. PMID- 11922173 TI - Detection of infectious agents in laboratory rodents: traditional and molecular techniques. AB - Methods to detect infectious agents in laboratory animals have traditionally been serological and culture based. Molecular methods to detect infectious agents in laboratory animals are being used more routinely. Confusion as to when and how to use molecular methods abounds. In this review, we present a guide to the weaknesses and strengths of using traditional and molecular methods for the detection of infectious agents in laboratory animals. PMID- 11922174 TI - Comparison of the deduced amino acid sequence of guinea pig adenovirus hexon protein with that of other mastadenoviruses. AB - PURPOSE: We sought to isolate, clone, and determine the nucleic acid sequence of the guinea pig adenovirus (GPAdV) hexon gene. From this, the amino acid sequence of the cloned portion was deduced and compared with a set of mastadenovirus hexons. METHODS: The DNA isolated from a histologic section of infected guinea pig lung was subjected to high-fidelity amplification, using degenerate primers complementary to a conserved nucleic acid sequence near the 3' end of the hexon gene of mastadenoviruses and a 5' primer from GenBank accession No. X95630 (GPAdV hexon gene partial sequence). The amplified product was cloned, the nucleic acid sequence was determined, and the amino acid sequence was deduced and compared with the hexon amino acid sequences of 25 mastadenoviruses. RESULTS: The cloned fragment comprised 1,603 base pairs (bp) [approximately 50%]) of the hexon. Of the initial 278 nucleic acids of the clone, 276 were identical with GenBank accession No. X95630, and the deduced amino acid sequences of both were identical. The deduced GPAdV hexon amino acid sequence from the clone aligned with structural regions NT, V1, DE1, and FG1 described for human adenovirus types 2 and 5. The GPAdV hexon had < 50% similarity in amino acid sequence, compared with hexons of 25 other mastadenoviruses. Analysis of regional peptide similarities revealed the GPAdV hexon to be more similar to animal mastadenoviruses and human subgroups A, C and F than to other human subgroups. CONCLUSIONS: The cloned portion of the GPAdV hexon contained a sequence nearly identical to that of GenBank accession No. X95630. Compared with the truncated amino acid sequences of human adenovirus types 2 and 5, the deduced GPAdV hexon amino acid sequence was similar in areas structurally conserved, but different in areas associated with type-specific antigenicity. PMID- 11922175 TI - Regional variations in the distributions of small intestinal intraepithelial lymphocytes (IELs) in BALB/c +/+, nu/+, and nu/nu mice. AB - Regional variations in intraepithelial lymphocytes (IELs) in the small intestine were examined in BALB/c +/+, nu/+, and nu/nu mice. The small intestine was obtained from 11- to 12-week-old mice and divided equally into three (proximal, middle, and distal) parts. The IELs were isolated from each part of the intestine, and the total numbers of IELs in nu/+ and nu/nu mice were about a fifth of those in +/+ mice. Regional variations in the distribution of the IEL alphabeta, but not the gammadelta T-cell subset were found by use of flow cytometry in +/+ and nu/+ mice. On the other hand, such differences were not found in nu/nu mice, suggesting that thymus-independent development of T cells is not different among regions. Different local expansion of thymus-dependent alphabeta T cells may cause the regional variations seen in the distribution of alphabeta T cell IELs in +/+ and nu/+ mice. PMID- 11922176 TI - Transoral intratracheal inoculation method for use with neonatal rats. AB - BACKGROUND AND PURPOSE: Studying the effects of toxic and infective compounds on the respiratory system requires a reliable method for delivering inoculum into the distal region of the lung. Although transoral intratracheal inoculation methods have been well documented for adult rats, to the authors' knowledge, a reliable method has not been validated for neonatal rats. The purpose of the study reported here was to develop a simple method for transoral inoculation in rat neonates. METHODS: Seven-day-old Fischer 344 rats were anesthetized with halothane, and a spinal needle was inserted in the tracheal lumen, by use of illumination and a modified otoscope. Meconium was injected into the lungs as a marker, and the neonates were kept under close observation. After euthanasia at 24 h, lungs were removed and fixed in formalin, and the microscopic distribution of the inoculum was assessed in the left, right cranial, middle, median, and caudal lung lobes. RESULTS: Microscopic examination of lungs indicated that intratracheal inoculation was achieved in 100% of neonatal lungs and the inoculum was consistently distributed in the alveoli of all pulmonary lobes. Important complications or mortality were not observed in the neonates. CONCLUSIONS: Intratracheal inoculation of neonatal rats is possible by use of a modified otoscope for transoral illumination. This technique is simple and reproducible and ensures, without complications, widespread distribution of inoculum in the lungs of neonatal rats. PMID- 11922177 TI - Three-dimensional magnetic resonance imaging of lung and liver tumors in mice by use of transversal multislice magnetic resonance images. AB - PURPOSE: To diagnose lung and liver tumors experimentally induced in mice in three-dimensional magnetic resonance (MR) images constructed by superimposing transversal multislice MR images of thoracic and abdominal regions taken under a high magnetic field of 7.05 tesla (T). METHODS: Lung and liver tumors were induced by administration of urethane to A/J mice and implantation of transplantable colon-26 cells into BALB/c mice, respectively. Two-dimensional (2 D) multislice MR images from the thoracic to abdominal regions were taken under the proton density-weighted conditions. Each organ in the 2-D MR images was pseudocolored, and a three-dimensional (3-D) image was constructed by superimposing them on a UNIX computer, using volume-rendering software. RESULTS: In the normal mouse, each organ in the thoracic and abdominal regions was three dimensionally imaged and was clearly distinguished from the others. In mice with tumors in the lung or liver, the pathologic changes in the tissue could be visualized in 3-D images. CONCLUSIONS: The MR images three-dimensionally constructed by use of a method combining MR imaging under a high magnetic field of 7.05 T and a computer technique using volume-rendering software was useful for diagnosis of lung and liver tumors experimentally induced in mice. PMID- 11922178 TI - Diagnostic polymerase chain reaction assays for identification of murine polyomaviruses in biological samples. AB - PURPOSE: Mouse polyoma virus and K virus are murine polyomaviruses frequently used in carcinogenicity and cellular biology studies in mice. These viruses can cause persistent infections, which increase the likelihood of transmission through transplantation of cells from infected mice. To identify polyomavirus infected biological samples, several diagnostic polymerase chain reaction (PCR) assays were developed. METHODS: Polyomavirus-family and virus-specific PCR assays were designed and optimized for specificity and sensitivity. The generic (polyomavirus-family) PCR assay and mouse polyoma virus-specific assays were compared with the mouse bioassay for diagnosis of infected cellular samples. RESULTS: Specificity of the PCR assays was confirmed by testing a battery of other murine viruses. The mouse polyoma virus PCR test was the most sensitive assay, detecting as few as 2,000 copies of homologous virus. The K virus PCR assay was about eightfold less sensitive, and the generic PCR test was the least sensitive. Mouse polyoma virus and generic PCR assays amplified mouse polyoma virus in the inoculum and tissues from experimentally infected mice, and performed better than did the mouse bioassay. CONCLUSIONS: Results of this study confirm that PCR is a specific and sensitive method for detection of murine polyomaviruses in biological samples. PMID- 11922179 TI - The obese Gottingen minipig as a model of the metabolic syndrome: dietary effects on obesity, insulin sensitivity, and growth hormone profile. AB - The objective of the study reported here was to induce obesity in the female Gottingen minipig to establish a model of the human metabolic syndrome. Nine- to ten-month-old female Gottingen minipigs received a high-fat high-energy (HFE) diet or a low-fat, low-energy (LFE) diet. The energy contents derived from fat were 55 and 13 %, respectively. After 5 weeks, animals were subjected to dual energy x-ray absorptiometry (DEXA) scanning, intravenous glucose tolerance testing (IVGTT), and 6-h growth hormone profile recording. After treatment, mean body weight of pigs of the LFE group was 21.0 +/- 0.4 kg, and was 26.8 +/- 0.2 kg in pigs of the HFE group (P < 0.0001). The DEXA scanning indicated that the fat content of the LFE group was 10.0 +/- 1.2 % versus 15.2 +/- 0.7 % in the HFE group (P < 0.003). Triglycerides concentration was significantly (P < 0.05) increased in pigs of the HFE group (0.24 +/- 0.03 mM), compared with that in pigs of the LFE group (0.13 +/- 0.04 mM). Preprandial plasma glucose and insulin concentrations were not affected, but insulin area under the curve during IVGTT was significantly high in the obese animals. Growth hormone (GH) secretion was low in both groups of pigs. The obese minipig shares some of the metabolic impairments seen in obese humans, and may thus serve as a model of the metabolic syndrome. PMID- 11922180 TI - Mamu-DQA1 allele and genotype frequencies in a randomly sampled breeding colony of rhesus macaques (Macaca mulatta). AB - We studied the allelic and genotypic distribution of the major histocompatibility class-II locus DQA1 observed in a random sample of Indian rhesus macaques (Macaca mulatta) from a major breeding facility in the United States. The DNA was isolated from whole blood samples collected between 1991 and 1994 from 65 Indian rhesus monkeys. Polymerase chain reaction-restriction fragment length polymorphism analysis (PCR-RFLP), which involves use of specific amplification of DQA1 exon 2 and subsequent restriction digestion of the 242-base pair fragment, was used to genotype the animals for the 20 known macaque (Mamu)-DQA1 alleles. Frequencies for four alleles (DQA1*240x, *2502, *2503 and *0102) differed significantly from those reported in a smaller sample of rhesus macaques from the German Primate Center. The modest genetic survey of Mamu-DQA1 genotypes presented here will be particularly useful in designing epidemiologic studies that investigate associations between immunogenetic background and disease susceptibility in macaque models of human disease. PMID- 11922181 TI - Intestinal cannulation: model for study of the midgut of the pig. AB - PURPOSE: To develop a pig model that would enable repeated biopsy specimen collection and endoscopic monitoring of the gut. This would increase precision of the experiment and reduce the number of experimental animals required. METHODS: Six 10-week-old Yorkshire pigs underwent surgery, and a cannula was inserted in the cecum. Two pigs served as non-operated controls. The health status of the animals was monitored by clinical, hematologic, and biochemical examinations and by studies of gut motility and microbial flora. The experimental period lasted for eight weeks and approximately 45 biopsy specimens were obtained from each animal. RESULTS: Repeated endoscopy was performed and biopsy specimens were taken. Adverse effects on the animal's health were not apparent, and differences were not evident in transit time of digesta or in diversity of the gut microbial flora. After surgery there was a transient increase in the concentrations of haptoglobin, serum amyloid A, and plasma cortisol, and in body temperature and white blood cell count. CONCLUSIONS: It is possible to use an intestinal cannula in the cecum both for endoscopy and biopsy specimen collection. The procedures did not influence health status of the pigs, nor alter gut function. The method will be useful in experimental infection studies as well as in other physiologic investigations. PMID- 11922182 TI - Atypical piscine mycobacteriosis in Japanese medaka (Oryzias latipes). AB - Japanese medaka, (Oryzias latipes), small, freshwater, tropical cyprinodonts, are principally used for toxicologic and carcinogenicity assays, but are finding more applications in developmental genetic and biological research. An increase in mortality began in brood stock of adult medaka that had been shipped and housed separately by sex. Initially, mortality averaged one fish daily and began in females two weeks after they were received. Cohabitation began eight weeks after arrival. After four to six weeks of cohabitation in different spawning aquaria, mortality was observed in males. Clinical signs of disease included loss of scale luster and color, with subsequent blanching of dorsal flank musculature, small raised nodules on various external surfaces, emaciation, fraying of fin tips, and equilibrium disturbances. Histologic examination of affected adults revealed multi-organ granulomatous inflammation with intracellular acid-fast bacilli. Specimens from 46 juvenile medaka that were spawned from affected adults, were submitted for culture and histologic evaluation. Of 18 fish, two had lesions similar to those of adults. The organism isolated from the remaining fish was identified as Mycobacterium fortuitum. Due to atypical rapid progression of disease, spread of M. fortuitum to progeny, and poor prognosis, the entire colony was euthanized. PMID- 11922183 TI - Spontaneous polar anterior subcapsular lenticular opacity in Sprague-Dawley rats. AB - A spontaneous focal polar anterior subcapsular lenticular opacity characterized by focal epithelial proliferation was found in Charles River Sprague-Dawley rats from various breeding facilities around the world (France, Japan, and the United States). The incidence of this change slightly increased with age up to a maximal incidence of 9.8% in 28- to 35-week-old male rats (French source). Over that period, there was little change in the size of the opacity; however some rats that were examined over longer periods (more than 2 years of age) developed secondary anterior cortical changes, and rarely, histologic findings of pigmentation and/or mineralization. The lenticular change was present throughout the life of the animals and had no sex predilection; mode of inheritance was not investigated. Due to its small size, this lens opacity is more easily identified by use of slit lamp biomicroscopy than by use of indirect ophthalmoscopy, and serial sections of the eye aid in locating it for histologic evaluation. PMID- 11922185 TI - Esophageal actinomycosis in a patient with AIDS. AB - Actinomycosis has been rarely reported in patients with HIV/AIDS in contrast to other opportunistic and common pathogens. We report a case of esophageal ulcer disease, secondary to actinomycosis occurring in a patient with recurrent odynophagia. The diagnosis was made histologically only after repeated upper endoscopy with biopsies. PMID- 11922184 TI - A voice that wraps around the body--communication problems in the advanced stages of non-small cell lung cancer. AB - INTRODUCTION: Significant problems in clinician-patient communication have been described in the oncology literatures. Advanced stage non-small lung cancer a devastating disease, can cause the communication between survivors, significant others, and clinicians to falter. To date, however, no studies have used qualitative methods to examine experiential aspects of living with non-small cell lung cancer. Nor have any studies evaluated the tools survivors might use to repair some of the damage caused by living with this disease. METHODS: Exploratory, two-part qualitative design. RESULTS: Survivors of non-small cell lung cancer live with multiple fears and losses. These include a diminished sense of self, the loss of health, fears of pain in a future tainted by the threat of death, and increased feelings of alienation due to the loss of previous sources of meaning in life. These experiences significantly affect cancer survivors abilities to communicate with clinicians and significant others. CONCLUSIONS: Survivors of non-small cell lung cancer often have difficulty sharing their experiences with others not suffering a similar affliction. Through their narratives with other survivors, however, patients are better able to initiate a biopsychosocial mechanism which enables them to create a cognitive map. This cognitive map helps survivors share their experiences with others, thereby repairing some of the damage caused by this disease, including the harm done to their communication with other people. PMID- 11922186 TI - Origins of cancer therapy. AB - This is a brief overview of the development of cancer therapy with a focus on systemic therapy. The modern era of chemotherapy developed at Yale University Medical School during World War II, a fact that has been generally unrecognized until recently. The observations preceding and involved in the discovery of effective drugs for cancer seem particularly pertinent for this anniversary year. PMID- 11922187 TI - Original memoirs: the control of bleeding in operations for brain tumors: with the description of silver "clips" for the occlusion of vessels inaccessible to the ligature. 1911. AB - One of the chief objects of concern in intracranial surgery should be the avoidance of any unnecessary loss of blood, for at best, in many cases of brain tumor associated with venous stasis, bleeding is likely to be so excessive as to necessitate postponement of the final steps of the procedure until a second or even a third session. The common methods of blood stilling by sponge, clamp, and ligature are largely inapplicable to intracranial surgery, particularly in the presence of bleeding from the nervous tissues themselves, and any device which serves as an aid to hemostasis in these difficult operations will bring a number of them to a safe termination at a single sitting, with less loss of blood and less damage to the brain itself. In addition to the more familiar tourniquet for the scalp, and wax for diploetic and emissary bleeding, suggestions are offered as to the use of gauze pledgets, dry sterile cotton, fragments of raw muscle and other tissues, as well as sections of organizing blood-clots for superficial meningeal bleeding, and silver "clips" for inaccessible individual points ether in dura or brain. The successful consummation of any critical operation often depends upon seeming trifles. It is, however, the scrupulous observance of surgical minutiae that makes possible the safe conduct of major intracranial performances--performances which a few years ago were attended in most cases by a veritable dance Macaber. PMID- 11922188 TI - Outbreak of bacterial conjunctivitis at a college--New Hampshire, January-March, 2002. AB - During February 1-14, 2002, approximately 100 students presented to a New Hampshire college's student health service with clinical signs of conjunctivitis (Figure 1). The cause of conjunctivitis was initially thought to be viral. However, because of the high number of cases, eye cultures were collected from 12 consecutive students; Streptococcus pneumoniae was isolated from cultures of all 12 students. The medical director of the student health service notified the New Hampshire Department of Health and Human Services about the outbreak and on February 22, the state health department requested assistance from CDC. This report summarizes preliminary results of the investigation of this outbreak, which indicate that an uncommon strain of pneumococcus caused this outbreak and that health-care providers should consider pneumococcus as a cause of conjunctivitis among college students. PMID- 11922189 TI - Update: allograft-associated bacterial infections--United States, 2002. AB - Tissue allografts are commonly used in orthopedic surgical procedures; in 1999, approximately 650,000 musculoskeletal allografts were distributed by tissue processors (1). A rare complication of musculoskeletal allografts is bacterial infection (2,3). After the reported death of a recipient of an allograft contaminated with Clostridium spp. (an anaerobic spore and toxin-forming organism) (3), CDC investigated this case and solicited additional reports of allograft-associated infections; 26 cases have been identified. This report summarizes the investigation of these cases and describes additional steps given to a tissue processor to enhance tissue transplant safety. PMID- 11922190 TI - Chagas disease after organ transplantation--United States, 2001. AB - Chagas disease (American trypanosomiasis) is caused by the protozoan parasite Trypanosoma cruzi. Chagas disease following solid-organ transplantation has occurred in Latin America, where Chagas disease is endemic, but has not been reported previously in the United States. This report describes three cases in the United States of T cruzi infection associated with transplantation of organs from a single donor. CDC and the U.S. organ transplantation organizations will consider whether to recommend screening of potential donors for T cruzi infection and, if so, which donors to screen, how to screen, and what to do if the screening tests are positive. PMID- 11922191 TI - Cat-scratch disease in children--Texas, September 2000-August 2001. AB - Cat-scratch disease (CSD), a bacterial infection caused by Bartonella henselae, has emerged as a relatively common and occasionally serious zoonotic disease among children and adults. To illustrate the spectrum of clinical manifestations of CSD observed during a 1-year period, Texas Children's Hospital (TCH) in Houston reviewed the medical records of 32 children evaluated at TCH during September 2000-August 2001 whose antibody titers indicated recent Bartonella infection. This report summarizes the evaluations of these cases and highlights four manifestations of infection with this pathogen in children. The findings emphasize that although CSD is generally a mild, self-limited illness, the differential diagnosis often includes more serious conditions (e.g., lymphoma, carcinoma, mycobacterial or fungal infection, or neuroblastoma) that might result in protracted hospital stays and lengthy treatments before diagnosis. Timely assessment of CSD is important, particularly when invasive diagnostic measures are being considered. PMID- 11922192 TI - Acquired rifamycin resistance in persons with advanced HIV disease being treated for active tuberculosis with intermittent rifamycin-based regimens. AB - Rifamycin drugs (i.e., rifampin, rifabutin, and rifapentine) are essential for short-course chemotherapy in persons with active tuberculosis (TB). However, adverse drug-drug interactions complicate the concurrent use of rifamycins and protease inhibitor drugs in persons with active TB who also are infected with human immunodeficiency virus (HIV-TB). CDC has recommended use of rifabutin in place of rifampin in multidrug regimens for the treatment of active TB in HIV-TB because rifabutin can be administered with antiretroviral treatment regimens that include protease inhibitors (1,2). These recommendations included twice-weekly intermittent therapy. Because intermittent rifabutin-based regimens had not been evaluated in clinical trials of HIV-TB, CDC's TB Trials Consortium (TBTC) initiated TBTC Study 23, a single-arm trial of twice-weekly rifabutin-based therapy for treatment of HIV-TB. PMID- 11922193 TI - Some thoughts about the etiopathogenesis of temporal arteritis--a review. AB - The three most striking features of temporal arteritis (TA) are the high incidence in populations of Nordic origins, the remarkable rarity of the disease among persons younger than 50 years, and the predilection for the superficial temporal arteries. At present, neither genetic susceptibility, environmental factors nor occurrence of specitic infections offer complete explanations to why Scandinavians are at increased risk of contracting TA. Similarly, recent developments in immunology and pathology of inflamed arteries cannot explain the preference of TA for this particular medium-size artery or why the disease almost exclusively affects persons older than 50 years. Although significant contributions regarding the pathogenesis of TA during the last decade are encouraging, the etiology of this disease still remains an enigma. PMID- 11922194 TI - Radiologic features poorly predict clinical outcomes in knee osteoarthritis. AB - OBJECTIVES: To assess the impact of radiographic severity and progression on pain and disability. METHODS: Measurements of mean joint space width (JSW), narrowest join space (NJS) point and assessment of symptoms by the WOMAC questionnaire were performed at baseline and after three years in 212 subjects over 50 years with primary knee OA. RESULTS: At baseline, JSW and NJS were not significantly correlated with the scores recorded for the WOMAC global index or its pain, stiffness or function subscales. A statistically significant correlation was observed between the joint space narrowing over three years and the changes observed in the pain subscale of the WOMAC during the same period. The three-year changes in the global WOMAC index in patients within the lowest and the highest quartiles of mean joint space width at baseline showed, in both cases, a statistically (p<0.05) significant favorable difference between patients treated with glucosamine sulphate and those having received placebo. CONCLUSION: Radiographic and clinical progressions of the disease are significantly associated but the clinical relevance of the association is questionable. PMID- 11922195 TI - Abnormal sonographic findings in the asymptomatic arthritic shoulder. AB - OBJECTIVE: To assess the diagnostic value of ultrasonography (US) in the evaluation of arthritic shoulder joints, especially in painless shoulders. METHODS: US examinations were performed in 57 consecutive patients with rheumatoid arthritis (114 shoulders) and in 32 controls (32 shoulders), using a 7.5 MHz linear probe and a standardized study protocol. US findings were compared with clinical, laboratory, and radiological data to find any relationship. RESULTS: Abnormal sonographic findings were found in 80 shoulders (70%); the most common were lesions in the supraspinatus tendon (38%), subacromial-subdeltoid bursitis (29%), bone erosions of the humeral head (20%), glenohumeral joint ellusion (19%), and biceps tendinitis (13%). Although US abnormalities were most frequent in patients with painful shoulders or abnormal findings on physical examination or radiography, a high rate of alterations was found in asymptomatic shoulders (51%), in normal shoulders on physical examination (44%) and in normal shoulders on radiographic assessment (61%). Differences of US findings in relation to time of evolution of rheumatoid arthritis, patient's age, and radiographic stage in hand and/or wrist joints were not found. CONCLUSION: US abnormalities in the shoulder joint are frequent in rheumatoid arthritis, both in patients with and without shoulder complaints as well as in patients with normal findings on physical examination. PMID- 11922196 TI - QT dispersion and cardiac involvement in patients with rheumatoid arthritis. AB - OBJECTIVE: To investigate the dispersion of repolarization variables in patients with rheumatoid arthritis (RA). METHODS: Electrocardiography (ECG) and Doppler echocardiography were performed on 40 patients with RA, which were divided into two groups according to the duration of disease and in 48 healthy controls. RESULTS: All patients had significantly longer QT dispersion (QTd) and corrected QT dispersion (QTc-d) values (p<0.05). The mean values of diastolic function variables were significantly different in all patients compared to healthy controls (p<0.05). There were no statistically significant differences between patient groups in terms of diastolic function variables except IVRT. However, QTd and QTc-d were significantly longer in patients with disease duration over 5 years (p<0.05). CONCLUSION: We conclude that repolarization heterogeneity and diastolic dysfunction are commonly seen in RA, and QTd is significantly longer in those patients with a disease duration over 5 years compared to those with new onset RA. PMID- 11922197 TI - Relationship between body mass index and fibromyalgia features. AB - OBJECTIVE: to evaluate the relationship between body mass index (BMI) and features of the fibromyalgia syndrome (FMS). METHODS: 211 female patients with FMS seen consecutively in our rheumatology clinic were analyzed. Spearman correlation was used. Further, FMS features were compared at different levels of BMI (kg/m2), e.g., < 25.00 vs > or = 25.00 (normal vs overweight). P value of < or = 0.01 was accepted as significant. RESULTS: A significant positive correlation was found between BMI and age (p<0.001) and a negative correlation between BMI and education (p<0.009). Health Assessment Questionnaire (HAQ) score was significantly correlated with BMI (p<0.001), whereas fatigue and number of tender points (TP) showed a trend (p=0.035 and 0.037, respectively). CONCLUSION: The HAQ score is significantly associated with BMI in FMS with a trend towards significance for fatigue and TP. Weight loss may improve physical functioning in this disorder. PMID- 11922198 TI - Evaluation of lipid profile, macular toxicity and clinical manifestations according to APO E genotype in systemic lupus erythematosus and rheumatoid arthritis patients treated with chloroquine. AB - OBJECTIVE: To investigate the effect of APO E gene polymorphism over lipid profile, macular toxicity and clinical manifestations in RA and SLE patients treated with chloroquine. MATERIALS AND METHODS: We studied 45 RA and 29 SLE patients treated with chloroquine who were classified based on the therapeutic regime of chloroquine into three groups: A) Cumulative dose of 100-300 g, B) >300 g and C) Never received chloroquine. Clinical evaluation, fasting lipid profile, visual field testing and stereoscopic photos of the retina were performed. APO E genotype was determined by PCR-RFLP. RESULTS: Reduced apo B levels in RA and SLE according to the cumulative dose of chloroquine 2/3 APO E genotype in a subset of SLE patients were observed. Macular toxicity was independent of both APO E genotype and cumulative chloroquine dose. CONCLUSIONS: Reduced apo B levels were observed associated to chloroquine treatment and 2/3 APO E genotype. PMID- 11922199 TI - Pro-inflammatory role of serotonin and interleukin-6 in arthritis and spondyloarthropathies--measurement of disease activity by bone scan and effect of steroids. AB - OBJECTIVE: As serotonin is a mediator of inflammatory joint disease, serum levels were investigated in human patients with arthritis for a possible corresponding role as a disease marker. DESIGN: 48 patients were evaluated by bone scan for disease activity. 5-HT and CRP were measured in the whole group, and IL-6 in those not yet receiving corticosteroids. The pro-inflammatory parameters were compared to each other and to scintigraphic features. RESULTS: The serum levels of serotonin did not correspond to disease activity measured by CRP, IL-6 or activity on joints in skeletal scintigraphy. No difference was seen in comparison to the values of a control group, but when glucocorticoid treatment was included, low 5-HT serum values were observed. A significant correlation between CRP and IL 6 as indicators of inflammation and bone scan results versus CRP could be shown. CONCLUSION: The measurement of serum serotonin provides no relevant information about disease activity in synovial inflammation. For monitoring osteoarthritis and synovial inflammation, bone scan and laboratory determination of CRP and IL-6 together appear to present useful information about infestation in the disease process. PMID- 11922200 TI - Pulmonary miliary tuberculosis in a patient with anti-TNF-alpha treatment. AB - No difference in the number of serious adverse events was reported in previous clinical trials in rheumatoid arthritis (RA) and inflammatory bowel disease (IBD) patients treated with TNF-alpha blockade, but a patient died because of disseminated tuberculosis. A tuberculosis reactivation in a patient with IBD and anti-TNF-alpha treatment has been recently reported. Very recently 70 cases of tuberculosis were reported from the FDA Adverse Event Reporting System. We report a case of pulmonary miliary tuberculosis in a RA patient treated with TNF-alpha blockade. The important role of TNF-alpha in defence against tuberculosis and possible mechanisms of anti-TNF-alpha agents impairing tuberculosis immune response are discussed. PMID- 11922201 TI - Measuring health related quality of life in patients with rheumatoid arthritis- reliability, validity, and responsiveness of a Swedish version of RAQoL. AB - OBJECTIVE: To adapt the Rheumatoid Arthritis Quality of Life (RAQoL) questionnaire for Swedish patients and evaluate psychometric properties in a prospective study. METHODS: Reliability was assessed in 61 patients filling in RAQoL two times with one week's interval. 114 patients completed RAQoL and Nottingham Health Profile (NHP) on 2-3 occasions 6 months apart. Validity was evaluated comparing RAQoL-scores to disease-related variables and NHP subscales. Standardized response mean was applied to calculate responsiveness with the RA related variables as external indicators of change. RESULTS: Test-retest reliability was high and internal consistency sufficient. RAQoL correlated as expected to NHP section scores. In a multivariate model the Stanford Health Assessment Questionnaire disability index (HAQ) and general health could explain 40% and disease activity measures 13% of the variance of RAQoL. Correlations between change scores of clinical variables and RAQoL and NHP were weak but positive. Standardized response means regarding change of disease activity, HAQ, and general health were small but in the same range for both RAQoL and NHP. CONCLUSION: The Swedish RAQoL had similar measurement properties as the original version. However, responsiveness regarding condition specific measures was not better than for the generic instrument NHP. PMID- 11922202 TI - A shifting landscape: what will be next FDG in PET oncology? AB - The tumor-seeking agent most widely used in positron emission tomography (PET) is 2-(18)F-fluorodeoxy-D-glucose (FDG). The clinical usefulness of FDG PET has already been proved in detecting, staging and restaging various kinds of malignant tumors, but nuclear medicine physicians suffer from a "diagnostic dilemma," in which a relatively high false positive ratio of FDG PET in diagnosing malignant tumors prevails. To increase more specific tumor uptake or more specific tumor characterization, numerous PET radiopharmaceuticals have been developed, and some of them are being tested in clinical trials. This review will briefly survey the tumor uptake mechanism and clinical significance of representative non-FDG PET radiopharmaceuticals used in clinical trials for patients with cancers. PMID- 11922203 TI - Performance evaluation of OSEM reconstruction algorithm incorporating three dimensional distance-dependent resolution compensation for brain SPECT: a simulation study. AB - Iterative reconstruction techniques such as an ordered subsets-expectation maximization (OSEM) algorithm can easily incorporated various physical models of attenuation or scatter. We implemented OSEM reconstruction algorithm incorporating compensation for distance-dependent blurring due to the collimator in SPECT. The algorithm was examined by computer simulation to estimate the accuracy for brain perfusion study. METHODS: The detector response was assumed to be a two-dimensional Gauss function and the width of the function varied linearly with the source-to-detector distance. The attenuation compensation (AC) was also included. To investigate the properties of the algorithm, we performed computer simulations with the point source and digital brain phantoms. In the point source phantom, the uniformity of FWHM for the radial, tangential and longitudinal directions was evaluated on the reconstruction image. As for the brain phantom, quantitative accuracy was estimated by comparing the reconstructed images with the true image by the mean square error (MSE) and the ratio of gray and white matter counts (G/W). Both noise free and noisy simulations were examined. RESULTS: In the point source simulation, FWHM in radial, tangential and longitudinal directions were 14.7, 14.7 and 15.0 mm at the image center and were 15.9, 9.83 and 10.6 mm at a distance of 15 cm from the center by using FBP, respectively. On the other hand, they were 8.12, 8.12 and 7.83 mm at the image center, and were 7.45, 7.44 and 7.01 mm at 15 cm from the center by OSEM with distance-dependent resolution compensation (DRC). An isotropic and stationary resolution was obtained at any location by OSEM with DRC. The spatial resolution was also improved about 6.5 mm by OSEM with DRC at the image center. In the brain phantom simulation, the blurring at the edge of the brain structure was eliminated by using OSEM with both DRC and AC. The G/W was 2.95 and 2.68 for noise free and noisy cases, respectively, when no compensation was performed. But the values for G/W without and with noise became 3.45 and 3.21 with AC only and were improved to 3.75 and 3.71 with both AC and DRC. The G/W approached the true value (4.00) by using OSEM with both AC and DRC even when there was statistical noise. CONCLUSION: In conclusion, OSEM reconstruction including the distance dependent resolution compensation algorithm was reasonably successful in achieving isotropic and stationary resolution and improving the quantitative accuracy for brain perfusion SPECT. PMID- 11922204 TI - 99mTc(V)-DMSA scintigraphy in monitoring the response of bone disease to vitamin D3 therapy in renal osteodystrophy. AB - Renal osteodystrophy (ROD) is a common and serious complication for uremic patients and patients are treated with 1,25-dihydroxyvitamin D3. The bone scanning agent 99mTc-phosphate has also been used to evaluate in ROD but it is not clear that bone scintigraphy has a role in the follow-up of treatment. In this study 99mTc(V)-DMSA scintigraphy was performed in eleven patients [age 40.7 +/- 17.3 (mean +/- SD) yr] with ROD before and after vitamin D3 therapy. Images were obtained after hemodialysis performed following tracer injection to maintain normal blood levels of the radiopharmaceutical and to reduce soft tissue activity. Lumbar vertebra-to-soft tissue uptake ratios (LUR) were quantified with the planar 99mTc(V)-DMSA images. Alkaline phosphatase and parathyroid hormone levels after treatment had significantly decreased compared with pre-therapy. In all patients there was visually decreased uptake in bone structures after treatment. After treatment the mean LUR ratio was significantly lower than those of before treatment (3.59 +/- 2.63 vs. 1.65 +/- 0.62; p = 0.01). LUR values were correlated with pre-therapy alkaline phosphatase and parathyroid hormone. These findings indicate that 99mTc(V)-DMSA scintigraphy is sensitive in evaluating the response of ROD to vitamin D3 therapy. PMID- 11922205 TI - Myocardial viability assessment with gated SPECT Tc-99m tetrofosmin % wall thickening: comparison with F-18 FDG-PET. AB - OBJECT: This study was designed to assess the value of gated SPECT Tc-99m tetrofosmin (TF) wall thickening (WT) in addition to TF exercise (Ex)/rest myocardial SPECT, in comparison with F-18 fluorodeoxyglucose (FDG)-PET. METHODS: The study population consisted of 33 patients with old myocardial infarction (27 men and 6 women; mean age, 62 +/- 8 years old). All patients underwent Ex/rest TF SPECT and glucose loading FDG-PET. Polar map images of Ex/rest TF were generated and divided into 24 segments for further analysis. We classified LV segments according to the exercise-rest perfusion scintigraphy. LV segments with less than 70% of the maximum TF activity on the exercise image were defined as stress induced defects. Among these, the segments whose TF activity increased by 10% from exercise to rest images or exceeded 70% of the maximum uptake were defined as reversible (viable) defects. The remaining defects on the rest image were irreversible (non-viable) defect segments, and were considered for viability study on the basis of %WT. %WT was calculated according to the standard method: [(counts ES - counts ED)/counts ED] x 100. A viable segment on gated SPECT was defined as a segment whose %WT exceeded the lower limit of the normal value (mean - SD). PET viability was defined as FDG uptake exceeding 50% of the maximum count. RESULTS: Among the 792 segments evaluated in the 33 patients studied, there were 689 PET viable segments. Of the 689 segments analyzed, 198 (29%) were identified as having defects on Ex images. Among these defects, 55 (8%) were reversible or partially reversible, as evidenced by rest images, and 143 (21%) were irreversible. Of the irreversible segments on Ex/rest images, 106 (15%) demonstrated no apparent WT by gated TF SPECT, whereas 37 (6%) segments with irreversible defects did have apparent WT. Overall, the sensitivity of Ex/rest TF perfusion imaging was 79%. Sensitivity was improved from 79% to 85% by combining %WT and perfusion data, but specificity was reduced from 70% to 56%. CONCLUSION: %WT evaluated from gated TF imaging enhanced myocardial viability assessment in comparison with FDG-PET. PMID- 11922206 TI - Serial change in 123I-MIBG myocardial scintigraphy in non-insulin-dependent diabetes mellitus. AB - PURPOSE: We performed 123I-MIBG (MIBG) myocardial scintigraphy twice in patients with non-insulin-dependent diabetes mellitus (NIDDM) to investigate whether MIBG distribution was improved by pertinent clinical control. To determine the influential factors for MIBG distribution, we investigated the association between various clinical parameters and the serial change in MIBG uptake parameters. PATIENTS AND METHODS: Twenty NIDDM patients with no cardiac disorders were evaluated. Planar images were taken at 30 minutes (early) and 3 hours (delayed) after MIBG injection. The heart-to-upper-mediastinum uptake ratio (H/M) and washout ratio (WR) were calculated as parameters for estimating cardiac sympathetic function. Patients were divided into two groups, eight in the improved group and twelve in the unimproved group, according to the serial change in H/M. The mean interval between the baseline and the follow up study was 2.1 +/ 0.6 year. Differences between the means of the laboratory data in patients in both groups were compared for the baseline and the follow up study by using the paired t-test. As a means of determining the influential factors for a serial change of MIBG uptake, Fisher's exact test was performed to evaluate the association between the serial change in cardiac MIBG parameters and changes in other clinical parameters, such as blood sugar (BS) control, BS control method (insulin therapy), serum cholesterol control, and severity of diabetic complications. We also analyzed the association between the changes in CV(R-R) (coefficient variance of R-R intervals at rest ECG) or NCV (velocity of posterior tibial nerve) and those of other clinical parameters. Associations among these neurological parameters (MIBG parameters, CV(R-R) and NCV) were also analyzed. RESULTS: Paired t-tests showed a significant decrease in fasting blood sugar and fructosamine in the improved group in the follow up study compared to those in the baseline study. Nevertheless, Fisher's exact test showed no significant association between FBS, HbA1C, fructosamine and the improvement in cardiac MIBG uptake. The only significant association was observed between the serial change in H/M and the BS-control method (insulin therapy). Within the neurological parameters, a significant association was noted between the serial changes in H/M and CV(R-R). CONCLUSION: Although BS control was likely to be an important factor, it did not always ameliorate cardiac MIBG uptake. Based on the significant association between the BS-control method (insulin therapy) and MIBG uptake change, the severity of diabetes mellitus was likely to be a more influential factor. It was suggested that cardiac MIBG uptake could improve within the mild stage if controlled by diet therapy or an oral hypoglycemic agent in NIDDM. PMID- 11922207 TI - Accuracy of plasma sample methods for determination of glomerular filtration rate with 99mTc-DTPA. AB - The plasma sample method following a single injection of radioactive markers has been proved to be simple and accurate for the determination of glomerular filtration rate (GFR) in clinical practice. The aim of this study was to assess clinical accuracy of single-, two- and multi-sample methods. METHODS: The study was performed on 50 patients with various degrees of renal dysfunction (29 males and 21 females; aged 27 to 90 years). As a reference the true GFR (GFRt) was determined by means of the two-compartment model curve fitting 10 plasma samples following a single-injection of 99mTc-DTPA. The GFRt was compared to the GFR estimated by the Christensen and Groth's single-sample (GFRcg), two-sample (GFR2s) and multi-sample (GFRm) between 75 and 300 min after the injection. The GFRs by two- and multi-sample methods were determined with the slope and intercept algorithm and its overestimation was corrected by Brochner-Mortensen's formula. RESULTS: In 49 patients with GFR between 12 and 169 ml/min/1.73 m2, the standard deviation of difference (95% limits of agreement) between GFRt and GFRcg at 180 min was 6.513 ml/min/l73 m2 (-16.5 approximately 9.5 ml/min/1.73 m2), which was somewhat closer than 7.311 ml/min/1.73 m2 (-12.5 approximately 16.5 ml/min/1.73 m2) in GFR2s in slow clearance phase at 120 min and 240 min. However, the single-sample method tended to show some scattering in GFR below 30 and above 140 ml/min/ 1.73 m2. On the contrary, the 2-sample method tended to be scattered in GFR above 120 ml/min/ 1.73 m2. CONCLUSION: In view of its accuracy and technical simplicity, the single-sample method is first choice in a routine practice. The two-sample method is essential of choice for a patient in whom the GFR is expected to be below 30 ml/min/1.73 mi2. These two methods may be chosen selectively in dependence on the preserved renal function which is expected at time of the test. PMID- 11922208 TI - The usefulness of dipyridamole thallium-201 single photon emission computed tomography for predicting perioperative cardiac events in patients undergoing non cardiac vascular surgery. AB - The aim of this study was to evaluate the usefulness of dipyridamole Tl-201 myocardium single photon emission computed tomography (201Tl-SPECT) for predicting perioperative cardiac events in patients with arteriosclerosis obliterans (ASO) and abdominal aortic aneurysm (AAA) undergoing non-cardiac vascular surgery. METHODS: Preoperative dipyridamole 201Tl-SPECT imaging in association with clinical risk assessment was performed in 224 consecutive patients (97 ASO and 127 AAA). RESULTS: The patients were classified into three groups, including low-risk (n = 173, 77%), intermediate-risk (n = 39, 18%), and high-risk (n = 12, 5%) groups according to the clinical risk stratification. The prevalence of reversible Tl-201 defect was significantly higher in the high-risk group than that in the low-risk group (83% vs. 14%, p < 0.001). In 180 patients who underwent vascular surgery, 9 patients (5.0%) had perioperative cardiac events, including heart failure (n = 1), unstable angina (n = 2), and other cardiac events such as arrhythmias (n = 6). The clinical variables including the clinical risk stratification did not significantly correlate with the perioperative cardiac events. In contrast, the reversible defect on 201Tl-SPECT was the only variable to predict perioperative cardiac events by a stepwise logistic regression analysis (odds ratio 7.0, 95% confidence interval 1.7-28.0, p = 0.007). It was also a significant predictor of perioperative cardiac events in a subgroup of low risk patients (odds ratio 11.6, 95% confidence interval 2.3 57.4, p = 0.004). The sensitivity and specificity of the reversible defect for predicting perioperative cardiac events were 55.6% and 84.8% in all operated patients, and 57.1% and 89.7% in low risk patients, respectively. CONCLUSIONS: The preoperative dipyridamole 201Tl-SPECT was useful for predicting perioperative cardiac events in patients with vascular diseases, even in patients identified as having a low risk based on the clinical risk assessment. PMID- 11922209 TI - New bone-seeking agent: animal study of Tc-99m-incadronate. AB - OBJECTIVE: Disodium cycloheptylaminomethylenediphosphonate monohydrate (incadronate disodium) is a third-generation bisphosphonate compound which potently inhibits bone resorption, and a highly effective drug in the treatment of metastatic bone disease. We first labeled incadronate disodium with 99mTc, and examined its biodistribution and bone uptake after intravenous injection in rats to assess its potential for clinical use as a bone-seeking agent for judgment of the therapeutic effect of incadronate on bone metastases. Bone scan with 99mTc labeled incadronate (99mTc-incadronate) may yield important information prior to the use of incadronate for treatment of bone metastases. METHODS: Synthesis of 99mTc-incadronate was carried out by reduction of 99mTc-pertechnetate in the presence of SnCl2 and N2 gas. Normal rats were injected with 18.5 MBq (0.5 mCi) 99mTc-incadronate in a volume of 0.1 ml intravenously and then sacrificed at 15 min, 30 min, 1 h or 2 h (six rats at each time point) after injection. Samples of muscle, stomach, small intestine, kidney, liver and bone (femur) were taken and weighed. In addition, a 1-ml sample of blood was drawn from the heart, and urine was taken from the urinary bladder immediately after sacrifice. Samples were measured for radioactivity and expressed as percent uptake of injected dose per gram or per milliliter (% ID/g or ml). Bone-to-blood and bone-to-muscle uptake ratios were determined from the % ID/g or ml values for these organs. RESULTS: The greatest accumulation of 99mTc-incadronate was found in bone. Radioactivity in bone was as high as 3.22 +/- 0.68% ID/g at 2 hours after injection. Scintigraphic images of 99mTc-incadronate in normal rats revealed highly selective skeletal uptake. CONCLUSION: 99mTc-incadronate exhibited high uptake in bone, and relatively low uptake in soft tissue, suggesting that it may be useful as a bone-seeking agent for judgment of the therapeutic effect of incadronate on bone metastases, by determining the degree of its accumulation in metastatic bone lesions. PMID- 11922210 TI - Coincidental visualization of an atypical bronchial carcinoid on Tc-99m-sestamibi scan in Kallmann's syndrome. AB - We report a case of an atypical bronchial carcinoid showing sestamibi uptake. A 27-year-old man with Kallmann's syndrome and hyperparathyroidism had parathyroid scintigraphy with 99mTc-sestamibi. A focal uptake was observed on the right perihilar region, and this right perihilar mass was demonstrated on computed tomography and proved histologically to be an atypical bronchial carcinoid tumor. Factors which may explain the tumoral avidity for sestamibi are increased blood flow, transmembrane potentials of plasma and mitochondrial membranes and the relative number of mitochondria present in the cells of this carcinoid tumor. The importance of this case is the coincidence of an atypical carcinoid in a patient with significant failure of secondary sexual characteristics, right renal agenesis and bilateral anosmia associated with Kallmann's syndrome. PMID- 11922211 TI - Usefulness of dynamic renal scintigraphy in a patient with renal arteriovenous fistula treated by transcatheter embolization. AB - Tc-99m-DTPA renal scintigraphy was performed before and after transcatheter coil embolization in a patient with renal arteriovenous fistula (AVF). Before embolization, scintigraphy showed a characteristic accumulation pattern compatible with the hemodynamics of an aneurysmal type of AVF in the vascular phase. After embolization, the effects of treatment, such as a change in the split renal function on the treated-side and the disappearance of the shunt flow through the fistula were evident. It was concluded that dynamic renal scintigraphy was very useful for patients with an aneurysmal type of renal AVF scheduled for transcatheter embolization. PMID- 11922212 TI - A case of general paresis showing marked treatment-associated improvement of cerebellar blood flow by quantitative imaging analysis. AB - We describe a patient with general paresis who developed progressive dementia and a cerebellar syndrome including wide-based gait, slurred speech, and intention tremor. Quantitative analysis by means of a Patlak plot of single-photon emission computed tomography (SPECT) with 99mTc-ethyl cysteinate dimer showed generally low blood flow in the cerebrum and the cerebellum. After antisyphilitic therapy, blood flow in the brain, especially in the cerebellum, improved noticeably, as did the cognitive disorder and the cerebellar syndrome. PMID- 11922213 TI - Radionuclide cisternography in intracranial hypotension syndrome. AB - A 44-year-old male patient complaining of severe headache right after twisting his body during aerobic exercises in a swimming pool underwent In-111 DTPA radionuclide cisternography. Leakage of cerebrospinal fluid (CSF) was proved on the right side of T2 and T3 vertebrae. Cisternography after bed and conservative treatments demonstrated the disappearance of abnormal tracer accumulations. Radionuclide cisternography is of great value in diagnosing cerebral-spinal fluid leak and in evaluating the therapeutic effect. PMID- 11922214 TI - Sialolithiasis. PMID- 11922215 TI - Acellular dermal graft: a human histologic case report. AB - It is the aim of this article to present a case report involving the biopsy and subsequent histologic analysis of an acellular dermal matrix. The area of the graft in contact with the root surface did not demonstrate a histologic attachment, but was defined as fibrous tissue apposition to the root surface. In addition, the coronal portion of the graft did not appear to be revascularized or revitalized as would be expected with an autogenous soft tissue graft. Finally, no new cementum formation was seen, but displacement of the junctional epithelium was observed. PMID- 11922216 TI - Progressive bone adaptation of titanium implants during and after orthodontic load in humans. AB - The aim of the present work was the evaluation of implant stability and periimplant bone reaction by histologic and clinical evaluation after therapeutic orthodontic loads. Forty-one adult patients received titanium implants as an orthodontic anchorage device; 12 patients received a retromolar or palatal implant to obtain tooth movement. Seven implants were removed at the end of the orthodontic therapy, after 2, 4, 6, and 12 months of orthodontic load, and processed for histologic examination. It was possible to distalize maxillary and mandibular molars and a group of teeth (molars and premolars), and to obtain tipping, uprighting, intrusion, extrusion, and transfer of anchorage in other parts of the mouth. The results showed that orthodontic therapy is facilitated and quickened by the use of implants. All implants remained stable in the bone up to 12 months of loading, and all were osseointegrated. Microfractures, microcracks, and microcalli were observed around implants that had been placed in both low- and high-density bone. The remodeling rate was still elevated after 18 months. PMID- 11922217 TI - Effect of platelet-rich plasma on bone growth and osseointegration in human maxillary sinus grafts: three bilateral case reports. AB - Platelet-rich plasma is an autologous product that is derived from whole blood through the process of gradient density centrifugation. The proposed value of this product in dental implantology and in bone augmentation procedures lies in the ability to incorporate high concentrations of the growth factors PDGF TGF beta1, TGF-beta2, and IGF as well as fibrin, into the graft mixture. Research has shown an increased bone maturation rate and improved bone density when this product, or its recombinant growth factors, is added to small bony defects or to larger defects that use autogenous bone as the grafting material. This study tested the efficacy of platelet-rich plasma in three bilateral sinus graft cases with grafts of anorganic bovine bone that contained minimal or no autogenous bone. Histomorphometric analysis indicated that the addition of platelet-rich plasma to the grafts did not make a significant difference either in vital bone production or in interfacial bone contact on the test implants. PMID- 11922219 TI - What does the future hold for periodontal tissue engineering? PMID- 11922218 TI - GTR for root coverage: a long-term follow-up. AB - The purpose of this study was to evaluate the long-term stability of the results obtained when guided tissue regeneration (GTR) is used to obtain root coverage. A mean root coverage of 92.3% was obtained at 6 months postoperative. However, at the final postoperative visit (mean 25.3 months), the mean root coverage was reduced to 58.8%. This difference was statistically and clinically significant. There was a statistically significant increase in recession (1.4 mm) and a statistically significant loss in attachment levels (1. 7 mm) between the 6-month and final postoperative evaluations. The results of this study question the long term stability of the result when GTR is used for root coverage. PMID- 11922220 TI - The ramp mattress suture: a new suturing technique combined with a surgical procedure to obtain papillae between implants in the buccal area. AB - This article has been written to show the opportunity and eventually the predictability to obtain new papillae between implants and a better esthetic result by the use of a new suturing technique. After raising a full-thickness flap from the palatal to the vestibular side, it can be stabilized in such a position using a new suturing technique (ramp mattress suture) to apply pressure and tearing forces on the flap in an apicocoronal direction at the vestibular site and an opposite traction in a coronoapical direction at the palatal site. The ramp mattress suture seems to be capable of pulling the flap in an apicocoronal direction in the vestibular site, as well as in a coronoapical direction in the palatal site. Thanks to such a mattress suture, it will be possible to obtain a more coronal gingival margin. After an adequate healing period of approximately 5 weeks, a vestibular scalloped gingivectomy is performed around the vestibular surface of the abutment to create either a scalloped gingival margin or interproximal papillae only in the vestibular area, forming a gingival ramp in a palatovestibular direction to reasonably reduce the residual increased vestibular depth and optimize the esthetic result. Eight patients, for a total of 56 papillae, were treated with this new suturing technique. The esthetic results satisfied both clinician and patient expectations. PMID- 11922221 TI - Surgical thickening of the existing gingiva and reconstruction of interdental papillae around implant-supported restorations. AB - With the present knowledge of osseointegration and implants, it seems reasonable to hope that implants will be placed in a way that provides optimal function, esthetics, and phonetics. The esthetic objectives of implant dentistry should be similar to those of conventional prosthodontics, and the esthetics for implants begins with implant placement. Often, because of limitations in position and/or quality of bone, implants may have to be placed in locations that are not ideal in the axial position. Preangulated and customized abutments have been used in some situations primarily to salvage poorly placed implants. An adequate zone of attached gingiva is also necessary around these restorations to hide the periimplant restoration boundary and provide for a good soft tissue profile, which is the essence of esthetics. There are numerous situations where implants have been placed in areas of minimal keratinized gingiva. The crown margin around these areas of thin gingiva is usually supragingival; therefore, the esthetic appearance is poor Three case reports, which are part of a preliminary study, outline a plastic surgical technique of tunneling/pouching and the use of submerged connective tissue grafts. This technique allows the thickening of the existing gingiva but is also useful for reconstructing interdental papillae around implant-supported restorations. PMID- 11922222 TI - A rare lesion of the periodontium: the gingival cyst of the adult--a report of three cases. AB - This article describes the clinical aspects and the histologic features of three cases of gingival cyst of the adult. The diagnostic problems of this rare entity are discussed. Radiographic examination to determine the presence of bone loss must be undertaken to differentiate the gingival cyst of the adult from the more frequent lateral periodontal cyst. PMID- 11922223 TI - Classification of altered dental esthetics. AB - Clinical procedures designed to reestablish the idealized framework for the achievement of optimal esthetic restorative results have been frequently reported. Yet complex cases, where remarkable improvements of esthetic appearance are achievable in spite of preoperative conditions, are infrequently presented because the presentation standards appear to be set for idealized situations. If a classification system that reflects dental esthetic compromises and ideals existed, cases could be categorized accordingly, and realistic expectations could be applied to assess the effectiveness and results of a course of therapy. Considering that there is not currently a published classification system to this end, it is the purpose of this article to tender a classification of altered dental esthetics. PMID- 11922224 TI - Early functional loading using Branemark dental implants. AB - The present review article discusses the one- and two-stage surgical protocol for dental implant placement as well as the critical amount of micromotion at the bone-implant interface to obtain proper osseointegration. The relevant literature supporting the hypothesis that "splinting of individual implants as soon as possible following installation via a rigid fixed device will most likely decrease the micromotion at the bone-implant interface thus facilitating proper bone healing (osseointegration)" is reported. As a consequence of this approach, the treatment period can be significantly reduced. Finally, the importance of an objective evaluation of the bone quality and initial implant stability is highlighted. The information thus obtained via tools already available will facilitate the decision as to whether to load implants immediately, early, or late, and the term "individual functional loading" is coined. PMID- 11922225 TI - Multidrug resistant salmonellosis: an escalating problem. PMID- 11922226 TI - Antimicrobial susceptibility pattern of Enterococcus species from urinary tract infections. AB - OBJECTIVE: To determine the extent of drug-resistance among Enterococcus species in, in vitro experiments. Test strains were isolated from cases with urinary tract infection. Antimicrobial sensitivity was determined with particular reference to the beta-lactams, glycopeptide and aminoglycoside groups of antibiotics. METHODS: Studies were carried out on pure cultures of enterococci isolated from urine specimens of 147 patients. Freshly voided, aseptically collected, midstream specimens of urine were cultured by using standard techniques. The identification and speciation of the strains of enterococci were performed by employing a battery of tests. In vitro drug-susceptibility tests of enterococci were performed against the following antibiotics: penicillin, ampicillin, ampi-sulbactum, co-amoxi-clavulanate, ciprofloxacin, vancomycin, teicoplanin, gentamicin and streptomycin, by employing the disk diffusion method. MIC values for penicillin, ampicillin, vancomycin and teicoplanin against enterococci were determined by the agar dilution technique. MIC determinations were further carried out by the E-test method, for the glycopeptide antibiotics. Prevalence of high level resistance to aminoglycoside antibiotics was determined by the agar dilution method. RESULTS: The most common species isolated was E. faecalis (87.07%), followed by E. faecium (10.88%) and E. durans (2.05%). The magnitude of resistance to both penicillin and ampicillin among the Enterococcus spp. was 23.13%, and that to ciprofloxacin was 55.78%. The three antibiotics: nitrofurantoin, co-amoxi-clavulanate, and ampi-sulbactum, showed resistance of 0.78%, 8.16% and 2.72%, respectively. High-level aminoglycoside resistance among strains of enterococci for streptomycin and gentamicin was found to be 33.84% and 36.92%, respectively. All the strains were susceptible to the glycopeptide antibiotics tested. CONCLUSION: The emergence of enterococci with alarming rates of resistance concomitantly to penicillins and aminoglycosides highlights the need for a more rational and restricted use of antimicrobials, in order to minimize the selection and spread of such strains. An early detection of these problem pathogens is also important for preventing any treatment failures. PMID- 11922227 TI - Squatting, blood pressure and stroke. AB - Most Indians still adopt squatting posture in toilets. In a group of 67 healthy volunteers, squatting produced a small (8.09 +/- 7.04 mm Hg) but significant rise in systolic blood pressure (SBP) but not in diastolic (DBP). However, in a group of randomly selected treated hypertensives (N=104) squatting produced a much greater and significant rise in both SBP (14.46 +/- 11.63 mm Hg) and DBP (9.10 +/ 9.19 mm Hg). The possible clinical significance of this rise of BP in squatting has been evaluated in 100 consecutive CT proved patients with stroke by analysing their stroke onset data in relation to time, place, posture and activity. Most strokes (52%) occurred in the morning hours (5 am-9 am) and at home (86%) and over a third (36%) while in toilets. Thirty six percent of strokes occurred when the subjects squatted, mostly during defecation. More than half of hemorrhagic strokes occurred in the squatting position. The relationship of these clinical observations with the BP changes noted above on squatting appears to be more than fortuitous. We would suggest that hypertensive subjects and those at risk of stroke should avoid squatting and urge physicians to check squatting BP while monitoring anti-hypertensive therapy. PMID- 11922228 TI - A comparative evaluation of the effects of oral lorazepam, alprazolam and diazepam on venous admixture. AB - OBJECTIVE: To compare the effects of oral diazepam, lorazepam and alprazolam premedication on venous admixture. MATERIAL AND METHODS: One hundred fifty patients divided in three groups were included in the study. The venous admixture was determined using the ISO-shunt nomogram. The values obtained 90 minutes after administration of the drugs were compared with the values before the drug administration. The Student's t-test was applied to find out the significance. RESULTS: These were highly significant change in increase in venous admixture (Qs/Qt) in group I patients 90 minutes after premedication as compared to premedication values. There was statistically insignificant difference in venous admixture (Qs/Qt) in group II and group III patients 90 minutes after premedication as compared to premedication values. CONCLUSION: From the present study it can be concluded that 2 mg of oral lorozepam given 90 minutes before surgery to healthy patients have significant effects on venous admixture. However, the effects of alprazolam and diazepam had no significant effect on venous admixture. PMID- 11922229 TI - A randomized placebo controlled trial of Inolter (herbal product) in the treatment of type 2 diabetes. AB - OBJECTIVE: To evaluate the safety and effectiveness of Inolter in achieving glycemic control and changes in lipid profile of newly diagnosed type 2 diabetics. MATERIAL AND METHODS: STUDY DESIGN: A randomized, placebo-controlled double blind. Sixty newly diagnosed patients of type-2 diabetes were randomized. Of these, 30 were put on Inolter monotherapy and 30 patients on placebo. Efficacy of Inolter was determined by observing glycemic control by estimating glycosylated haemoglobin (HbA1c) and fasting blood sugar (FBS), alteration of lipid profile by serum cholesterol, LDL, VLDL, HDL and triglycerides. STATISTICAL METHODS: Student's unpaired 't' test. RESULTS: Patients compliance was satisfactory and no adverse effects were observed. Better hypoglycemic effect was observed with Inolter both respect to fasting blood sugar (mean 65.4 +/- 63.72 mg/dl) compared to placebo (mean 26.45 +/- 38.3 mg/dl) (p < 0.001). The mean change, with Inolter, in serum cholesterol (21.33 +/- 26.05 mg/dl), serum triglyceride (30.36 +/- 32.62 mg/dl), VLDL (8.85 +/- 3.25 mg/dl), LDL (10.00 +/- 4.48 mg/dl) and HDL (-2.33 +/- 5.66 mg/dl) was also significantly different (p < 0.001) from those given placebo, where it was 15.7 +/- 20.14 mg/dl, 8.70 +/- 13.24, 0.10 +/- 4.10 mg/dl, -0.70 +/- 4.58 mg/dl and 0.65 +/- 0.00 mg/dl, respectively. CONCLUSION: Inolter appears to be an useful adjunctive therapy to exercise and diet control in newly diagnosed type 2 diabetes mellitus. PMID- 11922230 TI - Evaluation of clinical criteria for the diagnosis of leptospirosis. AB - OBJECTIVES: Leptospirosis has a wide range of clinical presentation and therefore, clinical suspicion of the infection is often difficult. The objective of this study is to find out the usefulness of the clinical and epidemiological criteria in the diagnosis of leptospirosis and its comparison with microagglutination test (MAT). METHODS: A total of 118 patients with undiagnosed fever of more than seven days duration were included in the study. Their clinical presentation was scored on the basis of a clinical criteria. Sera of the patients were tested for antibodies against leptospira with the help of microagglutination test using a battery of antigens. The usefulness of the criteria was evaluated and compared with microagglutination test. RESULTS: A total of 44 out of 118 (37.28%) patients could be provisionally diagnosed as cases of leptospirosis on the basis of the clinical criteria. Eighteen of these 44 (40.9%) patients had serological evidence of leptospirosis. The criteria had a sensitivity of 81.81%, specificity of 72.91%, a positive predictive value of 40.9% and a negative predictive value of 94.59% when compared with microagglutination test. CONCLUSIONS: The criteria had a moderate sensitivity and specificity. Considering the non-specific signs and symptoms of this infection, the positive predictive value is significantly high. The criteria has a high negative predictive value and this would help the clinicians exclude the diagnosis of leptospirosis with precision. PMID- 11922231 TI - Comparative evaluation of market spacer and home made spacer in the management of bronchial asthma. AB - INTRODUCTION: A study was conducted to compare the efficacy of market available spacer (with valve) and home made spacer (without valve)--Bislery bottle. MATERIAL AND METHODS: Fifteen patients of bronchial asthma were included in the study. With the use of both devices there was significant bronchodilator effect. The reversibility using Bislery bottle was same as with spacer (market available) while comparing the FVC, FEV1, FEV1/FVC %, FEF 25-75 and PEFR value. RESULT: The difference in percent change in reversibility values by both the devices was not statistically significant (p < 0.05). CONCLUSION: We concluded that the Bislery bottle (without valve) is very cheap compared to market-available spacer and is equally effective which, therefore, can be substituted in bronchial asthma patients, who are unable to afford the cost of market available spacers. PMID- 11922232 TI - Inhibition of membrane Na+-K+ ATPase activity: a common pathway in central nervous system disorders. AB - OBJECTIVES: The study was conducted to assess the role of hypothalamic digoxin in neuropsychiatric and systemic disorders. A hypothesis regarding the central role of hypothalamic digoxin in neuroimmunoendocrine integration is proposed. METHODOLOGY: Blood samples from patients of CNS glioma, multiple sclerosis, systemic lupus erythematosis, subacute sclerosing panencephalitis, primary generalized epilepsy, Parkinson's disease, Down syndrome, AIDS dementia with neuropsychiatric features, syndrome X with multiple lacunar state, senile dementia, familial group (a family with familial coexistence of schizophrenia, Parkinson's disease, primary generalized epilepsy, malignant neoplasia, rheumatoid arthritis and syndrome X over three generations), schizophrenia and manic depressive psychosis were analysed for RBC membrane Na+-K+ ATPase, levels of digoxin and Mg++. RESULTS: Inhibition of RBC membrane Na+-K+ ATPase activity was observed in most cases along with increase in the levels of serum digoxin and decrease in the level of serum Mg++. CONCLUSION: The decreased Na+-K+ ATPase activity can be due to increased digoxin, which is a potent inhibitor of this enzyme. The inhibition of Na+-K+ ATPase can contribute to increase in intracellular calcium and decrease in magnesium, which can result in 1) defective neurotransmitter transport mechanism, 2) neuronal degeneration and apoptosis, 3) mitochondrial dysfunction, 4) defective golgi body function and protein processing dysfunction, 5) immune dysfunction and oncogenesis. The mechanism of how increased intracellular calcium and decreased magnesium can contribute to the above effects is discussed. PMID- 11922233 TI - Current concepts in the management of acute ischemic stroke. AB - The treatment of stroke is multifaceted, and is governed by three, well established goals: 1) To minimize the extent of brain; 2) To medically support the stroke patient; and 3) To prevent further brain injury secondary to initial event or repeated vascular insults. Certain guidelines for emergency care, based on clinical experience and knowledge of pathophysiology, have been accepted in many institutions as the standard. This article provides guidelines about the current management of acute ischemic stroke based on currently available data from clinical trials. The target audiences for this article are emergency room physicians and neurologists, who manage the patients during the first few hours after stroke. PMID- 11922234 TI - Human strongyloidiasis in AIDS era: its zoonotic importance. AB - Human strongyloidiasis is caused by a nematode Strongyloides stercoralis. Many species cause strongyloidiasis in animals. The parasite has predilection to one host only but the host specificity is not strict. When animal species infects humans there is intense allergic reaction in the form of cutaneous larva currens and larva migrans. Therefore, strongyloidiasis in strict terms is a zoonotic disease. The strongyloides species have three stages. The parasitic form inside the host, the free form stage in the soil or water that moults to infective third stage. The later infects the host through skin and migrate to the heart and lung and finally swallowed back to cause intestinal infection. However, in some cases intense pulmonary manifestations may take place. The Strongyloides stercoralis has unique feature of moulting from parasitic form to infective stage within the body, rather than coming out and forming free living stage and causing autoinfection. This may lead to latent infection for indefinite period in an immunocompetant person but fatal hyper or disseminated infection in immunocompromised person like patients of AIDS, organ transplant recipients, cancer and other patients put on immunosuppressive therapy, in whom it can involve any organ of the body. Because this group of patients in last few years have increased tremendously in Africa and South-East Asia, more and more cases of strongyloidiasis are being reported in english literature. The diagnosis of intestinal strongyloidiasis is made by repeated stool smear examinations and in extraintestinal strongyloidiasis the appropriate specimen is examined for the rhabditiform larvae. Recently serological tests have also been developed that can be used for epidemiological purposes. The drug of choice for the treatment of strongyloidiasis remains thiabendazole but due to its unacceptable side effects other medicines like albendazole and ivermectine are being used more frequently. The prevention of the infection is possible by adopting good personal hygiene and safe drinking water supply. PMID- 11922235 TI - Health care delivery in India today. PMID- 11922236 TI - Ciprofloxacin-induced QTc prolongation. AB - Ciprofloxacin has been widely used for treating infections and has been found to have very low cardiovascular side effects. QTc prolongation with the use of ciprofloxacin is yet to be reported in literature. A case report highlighting QTc prolongation by use of ciprofloxacin is being presented. PMID- 11922237 TI - Quadriplegia complicating ossification of the posterior longitudinal ligament. AB - We report a case of quadriplegia complicating ossification of posterior longitudinal ligament (OPLL) in a patient who was also found to have diffuse idiopathic skeletal hyperostosis (DISH). She also had osteomalacia (Vit. D deficiency) with secondary hyperparathyroidism. There could be a cause and effect relationship between the abnormal biochemistry and OPLL. PMID- 11922239 TI - Myopathy secondary to simvastatin in a type 2 diabetic patient. AB - The most important but rare adverse effect of simvastatin is myopathy. In megatrials with simvastatin, the overall incidence of myopathy is 0.025%. We present a case of myopathy presenting as proximal muscle weakness in both upper limbs secondary to simvastatin which reversed spontaneously after cessation of the drug. PMID- 11922238 TI - Jean Corvisart (1755-1821) first day cover and stamp of France 1964. PMID- 11922240 TI - Linear scleroderma and autoimmune hemolytic anaemia. AB - Coexistence of localized scleroderma with other autoimmune disorders is not seen frequently. It has been reported to occur with myasthenia gravis, hemiatrophy and systemic lupus erythematosus. In this report we describe an association wherein linear scleroderma coexisted with autoimmune haemolytic anaemia. PMID- 11922241 TI - Respiratory failure in acute intermittent porphyria. AB - We report two patients of acute intermittent porphyria (AIP) who presented with acute respiratory failure. Only one such previous report could be found. Occasionally, neuropathy may be the presenting feature in AIP which may progress to respiratory embarrassment. The cause of this neuropathy has been hypothesized to be direct neurotoxicity of delta-ALA by interaction with GABA receptor, altered tryptophan metabolism and may be heme depletion in nerve cells. PMID- 11922242 TI - Sleep disorders versus epilepsy: use of video EEG as a diagnostic tool. AB - Sleep disorders, especially parasomnias, are easily confused with the nocturnal epilepsies. Two cases are presented to show how misdiagnosis can occur, and video electroencephalogram can be used to resolve the issue so as to prescribe the specific treatment, avoid wrong treatment and unnecessary suffering. PMID- 11922243 TI - Cerebrotendinous xanthomatosis. PMID- 11922244 TI - Acute pituitary necrosis following snake envenomation. PMID- 11922245 TI - Life threatening stridor due to bilateral recurrent laryngeal nerve palsy as an isolated manifestation of intermediate syndrome. PMID- 11922246 TI - Burkitt's lymphoma. PMID- 11922247 TI - Who will bell the cat? PMID- 11922248 TI - Bruxism as presenting feature of Parkinson's disease. PMID- 11922249 TI - Coomb's negative autoimmune hemolytic anemia with thrombocytopenia (Evan's syndrome). PMID- 11922250 TI - Relapse pattern of Plasmodium vivax in Mumbai: a study of 283 cases of vivax malaria. PMID- 11922251 TI - Isoniazid chemoprophylaxis in Indian scenario: need for a practical approach? PMID- 11922252 TI - Filarial lymphoedema and alpha-1-adrenergic receptor blockade. PMID- 11922253 TI - Clinical practice guidelines on bronchial asthma in adults. AB - Effective management of asthma requires a partnership between the patient and the health care provider. This depends on the patient having a good understanding of the disease and its management. As the level of literacy is widely variable in India, patient education methods may have to be tailored according to the patient's ability to understand. In any case, patients with asthma should be able to a) Understand that asthma is a chronic disease requiring prolonged treatment b) Understand that treatment for asthma should be continued even when symptoms have abated c) Understand that control of asthma needs to be assessed by using objective methods periodically e.g. PEFR measurement d) Recognise when asthma control is deteriorating e) Take appropriate steps when asthma control deteriorates f) Understand the basic nature of the drugs they need to take e.g. preventers, relievers, and protectors g) Recognise the advantages of the inhaled route over the oral route h) Take steps to avoid "triggers" which can aggravate or precipitate an asthma attack. Patients who are more literate can be instructed further on drug interactions, safety, efficacy and side effects of steroids as well as the influence of other diseases on asthma and its treatment. PMID- 11922254 TI - Acellular dermal matrix allografts to achieve increased attached gingiva. Part 2. A histological comparative study. AB - BACKGROUND: In part 1 of this study, we compared the clinical efficacy of freeze dried acellular dermal matrix (ADM) allograft in 6 patients with autogenous free gingival graft (FGG) in 6 patients for increasing the width of attached gingiva in the mandibular anterior area. The purpose of the present study was to histologically compare the microstructure of ADM and FGG treated sites from the same group. METHODS: Biopsies were harvested from all 12 patients at 6 months postsurgery. The biopsies included the grafted sites with adjacent alveolar mucosa and gingiva propria and also donor palatal mucosa saved at the time of surgery. The 5 microm thick, neutral buffered formalin fixed, paraffin-embedded tissue sections were stained with hematoxylin and eosin (H&E), Masson's trichrome, and Verhoeff-van Gieson stains in order to investigate the density of collagen and elastic fibers. Additional sections were stained with periodic acid Schiff (PAS) and Papanicolaou's stain to identify the presence of glycogen granules in the epithelial layer and to highlight the keratin layer respectively. RESULTS: The unique appearance of ADM-derived tissue did not parallel any known oral mucosa. The connective tissue portion contained dense to extremely dense collagen fibers along with scattered elastic fibers. The demarcations between the ADM graft and the coronal gingiva as well as the apical alveolar mucosa were usually not very defined. A moderate to thin epithelial layer, with heterogeneous expression of keratinization and flat epithelium-connective tissue interface, covered the lamina propria. Both the thickness of the epithelium and the degree of keratinization decreased in apical direction, being mostly para- or orthokeratinized in the area close to gingiva and non-keratinized adjacent to the alveolar mucosa. In the FGG-treated sites, the density of collagen fibers was less than in ADM-derived tissue, palatal mucosa, and gingiva. Elastic fibers were very sparse, comparable to gingiva, but much less than in ADM-derived tissue. The epithelium was moderate, somewhat thinner but the shape of the rete ridges resembled that of palatal mucosa. Similar to gingiva epithelium, the epithelium of the FGG-treated area was relatively uniform in both thickness and keratinization, mostly para-keratinized with a well defined border to the non keratinized alveolar mucosa. Underneath the FGG-alveolar mucosa junction, a scar band composed of extremely dense collagen fibers consistently existed. CONCLUSIONS: The results of this 6-month histological evaluation suggest that: 1) the resultant tissue types of ADM grafts were similar to "scar" tissue; 2) the non-vital dermal matrix of ADM allograft lacked the capability of directing cyto differentiation of the covering epithelium; 3) autogenous FGG-derived tissue was neither identical to donor palatal mucosa nor to adjacent gingiva propria; 4) the connective tissue of donor palatal mucosa only partially contributed to the differentiation of the epithelium covering the FGG-treated area; and 5) the epithelium/connective tissue microenvironment surrounding the recipient site influenced the epithelial differentiation of the graft; this may play a more critical role in ADM grafting than in the grafting of autogenous FGG. PMID- 11922255 TI - Histomorphometrical analysis of bone formed after maxillary sinus floor augmentation by grafting with a combination of autogenous bone and demineralized freeze-dried bone allograft or hydroxyapatite. AB - BACKGROUND: Maxillary sinus floor augmentation procedures are currently the treatment of choice when the alveolar crest of the posterior maxilla is insufficient for dental implant anchorage. This procedure aims to obtain enough bone with biomaterial association with the autogenous bone graft to create volume and allow osteoconduction. The objective of this study was to histologically and histometrically evaluate the bone formed after maxillary sinus floor augmentation by grafting with a combination of autogenous bone, from the symphyseal area mixed with DFDBA or hydroxyapatite. METHODS: Ten biopsies were taken from 10 patients 10 months after sinus floor augmentation using a combination of 50% autogenous bone plus 50% demineralized freeze-dried bone allograft (DFDBA group) or 50% autogenous bone plus 50% hydroxyapatite (HA group). Routine histological processing and staining with hematoxylin and eosin and Masson's trichrome were performed. RESULTS: The histomorphometrical analysis indicated good regenerative results in both groups for the bone tissue mean in the grafted area (50.46+/ 16.29% for the DFDBA group and 46.79+/-8.56% for the HA group). Histological evaluation revealed the presence of mature bone with compact and cancellous areas in both groups. The inflammatory infiltrate was on average nonsignificant and of mononuclear prevalence. Some biopsies showed blocks of the biomaterial in the medullary spaces close to the bone wall, with absence of osteogenic activity. CONCLUSIONS: The results indicated that both DFDBA and HA associated with an autogenous bone graft were biocompatible and promoted osteoconduction, acting as a matrix for bone formation. However, both materials were still present after 10 months. PMID- 11922256 TI - Distraction osteogenesis and histogenesis in beagle dogs: the effect of gradual mandibular osteodistraction on bone and gingiva. AB - BACKGROUND: No study has systematically evaluated the effect of distraction osteogenesis on the gingival tissues. Therefore, this study was designed to analyze the newly formed bone and gingiva during the consolidation period of mandibular osteodistraction using standard histologic techniques. METHODS: Seventeen skeletally mature male beagle dogs underwent 10 mm of bilateral interdental mandibular lengthening. After distraction, the regenerates were allowed to consolidate for 0, 2, 4, 6, or 8 weeks, then the animals were sacrificed and tissues harvested for analysis. RESULTS: Mineralization began at the host bone margins at the end of the distraction period, followed by a progressive increase in bone surface area, with a concomitant decrease in fibrous tissue. The gingiva initially underwent mild inflammatory and reactive changes during distraction and during the first few weeks of consolidation. The rate of bone formation gradually increased from the end of distraction to the fourth week of consolidation, at which time it remained constant until sometime before the eighth week, when it tapered off slightly as remodeling began. From the second through the eighth week of consolidation, regenerative changes and neohistogenesis were seen in the gingival tissues. CONCLUSIONS: Osteodistraction has the potential to drastically decrease the total treatment time for alveolar bone augmentation prior to dentoalveolar implant placement since the regenerate bone rapidly mineralizes within approximately 8 to 10 weeks after the distraction period and the gingiva responds favorably to increased length by regeneration rather than by degeneration. Although the results appear favorable, similar data should be evaluated in human clinical trials. PMID- 11922257 TI - Periodontopathic bacteria in young healthy subjects of different ethnic backgrounds in Los Angeles. AB - BACKGROUND: The present study determined risk indicators for oral colonization by Actinobacillus actinomycetemcomitans, Porphyromonas gingivalis, Bacteroides forsythus, and Treponema denticola in 150 children and adolescents, 4 to 16 years of age, living in Los Angeles, California. METHODS: Fifty Caucasians, 50 Hispanics, and 50 Asian-Americans completed a questionnaire on demographic characteristics. 16S rRNA-based polymerase chain reaction identification was employed to determine the presence of test bacteria in unstimulated saliva. Step wise logistic regression analysis identified explanatory variables (risk indicators) accounting for the salivary presence of periodontopathic bacteria. RESULTS: A. actinomycetemcomitans occurred in 15%, P. gingivalis in 15%, B. forsythus in 14%, and T. denticola in 18% of all subjects. Two or more pathogens were detected in 20% of Hispanic subjects and in 12% of Asian-American subjects but not in any Caucasians (P = 0.0005, chi square test). However, no stable multivariate model including ethnicity was found for multiple pathogens. Risk for harboring any pathogens increased with the length of time lapse from last dental visit (odds ratio [OR], 4.46; 95% confidence interval [CI], 1.83 to 12.21), and decreased with higher education level of the mother (OR, 0.258; 95% CI, 0.052 to 0.875). Risk for harboring 2 or more periodontal pathogens decreased with the years the parents had resided in the United States (OR, 0.95; 95% CI, 0.901 to 0.992). Risk for harboring A. actinomycetemcomitans decreased as the number of years the parents had resided in the United States increased (OR, 0.91; 95% CI, 0.86 to 0.95), and decreased with higher income level of the father (OR, 0.201; 95% CI, 0.038 to 0.948). Girls were at higher risk for harboring P. gingivalis (OR, 2.55; 95% CI, 1.02 to 7.03), but at lower risk for carrying T. denticola (OR, 0.42; 95% CI, 0.17 to 0.98). CONCLUSIONS: This study showed that salivary occurrence of periodontopathic bacteria in young individuals was related to the length of time the parents had lived in the United States, education level of the mother, length of time since last dental visit, and gender, but apparently not to ethnicity per se. PMID- 11922258 TI - Tongue piercing: impact of time and barbell stem length on lingual gingival recession and tooth chipping. AB - BACKGROUND: The increasing popularity of tongue piercing has prompted several case reports documenting oral complications of this practice. However, there are no studies assessing potentially significant parameters. The purpose of this study was to evaluate the effect of time (years of wear) and tongue barbell size (stem length) on gingival recession and tooth chipping. METHODS: Fifty-two adults (mean age 22) with tongue piercings were examined for gingival recession on the lingual aspect of the 12 anterior teeth and for tooth chipping anywhere in the mouth. Subjects were grouped according to years of wear (0 to 2, 2 to 4, and 4+ years) and barbell stem length (long > or =1.59 cm, or short <1.59 cm). Data analysis was based on binomial test and non-parametric tests. RESULTS: No subject with a tongue piercing <2 years (group 0-2) exhibited lingual recession or tooth chipping. Lingual recession was found on mandibular central incisors in 50% of subjects wearing long barbells for 2 or more years. Tooth chipping was found on molars and premolars in 47% of subjects with a tongue piercing for 4+ years. CONCLUSIONS: Tongue piercing is associated with lingual recession of mandibular anterior teeth and chipping of posterior teeth. Long-term use of a tongue barbell increases the prevalence of these complications. Barbell stem length appears to differentially affect prevalence of recession and chipping. Since the overwhelming majority of subjects with tongue piercings are young adults, cessation efforts are needed to target this population. PMID- 11922259 TI - Relationships between clinical attachment level and spine and hip bone mineral density: data from healthy postmenopausal women. AB - BACKGROUND: There are physiological reasons to expect an association between bone mineral density of the spine and hip and attachment loss. To this point, however, most studies have found no correlation. METHODS: The 135 patients in this report were part of a randomized controlled trial of estrogen replacement. All patients were in good oral health at entry and received annual oral prophylaxis as part of the study. Standard probing measurements were made with a pressure sensitive probe at 6 sites on each tooth. Bone mineral density was measured with dual energy x-ray absorbtiometry at the lumbar spine (anterior-posterior and lateral) and proximal femur (neck, trochanter, intertrochanter, Ward's triangle, and total area). These procedures were performed at baseline and at annual intervals for 3 years. RESULTS: Correlations between cross-sectional measurements of clinical attachment level and bone mineral density were very weak, and did not approach statistical significance (-0.06 < or =r < or =0.10, 0.15 < or =P < or =0.75). A few somewhat stronger correlations were found between longitudinal changes in bone mineral density and attachment (-0.20 < or = r < or =-0.02, 0.02 < or = P < or =0.81). Although the correlations in the longitudinal changes were weak, they were consistently in the direction of greater bone mineral density being associated with less attachment loss. CONCLUSIONS: There is no clear association between clinical attachment level and bone mineral density of the lumbar spine and proximal femur, whether examined on a cross-sectional or longitudinal basis. Patterns in the data suggest there may be a weak association in the longitudinal changes. PMID- 11922260 TI - A method to assess the clinical outcome of ridge augmentation procedures. AB - BACKGROUND: Alveolar ridge augmentation procedures are often needed prior to implant placement or prosthetic rehabilitation. The purpose of this study was to develop a quantitative method to assess the clinical outcome of such procedures. METHODS: Two volumetric methods, based on the volume difference between the pre- and postoperative ridge, were developed. Either a transparent vacuum stent material (method A) or a silicone impression material in a custom acrylic tray (method B) was used to make an impression of the postoperative stone cast. This impression was then filled with polyvinylsiloxane bite registration material (PBRM) and placed on the preoperative cast. The volume of PBRM at the site of the augmented ridge (corresponding to the achieved ridge volume increase, in ml) was measured gravimetrically. Two independent examiners used both methods, in random order, to assess the outcome of 6 diverse clinical cases. Triplicate measurements were made (method precision). Method accuracy was assessed by fabrication of metal castings (known volumes) applied on a preoperative cast to simulate an augmentation procedure. RESULTS: Intra- and inter-examiner reproducibility was high (intraclass correlation coefficients > or =0.84) for both methods. The volumetric measurements obtained by method B showed excellent correlation with the predetermined augmentation volumes (r2 = 0.99; y = 0.94x + 0.12), in contrast to the measurements obtained by method A (r2 = 0.46; y = 0.43x + 1.37). CONCLUSIONS: The developed volumetric method (method B) is both precise and accurate in assessing the outcome of ridge augmentation. This simple, cost effective, and easy to implement method should be helpful in clinical studies of ridge augmentation procedures. PMID- 11922261 TI - A novel mutation of the cathepsin C gene in Papillon-Lefevre syndrome. AB - BACKGROUND: Papillon-Lefevre syndrome (PLS) is a disorder that involves destruction of the periodontium and abnormal hyperkeratosis of the palms of the hands and soles of the feet. Mutations of the lysosomal protease cathepsin C gene (CTSC) have been associated with PLS. However, genotypic and phenotypic correlation has not been established. In the present study we investigated the CTSC gene in a Brazilian cohort affected by PLS. METHODS: Eight consanguineous members of a kindred with PLS were studied. DNA was extracted and all exons of the gene amplified by the polymerase chain reaction (PCR) using specific primers. Mutations were identified by DNA sequencing of the coding region and introns of the CTSC gene. RESULTS: Sequence analysis of CTSC from subjects affected by PLS identified a novel mutation (587T --> C) in exon 4, predicted to cause a Leu196Pro amino acid substitution. Three of 3 subjects were homozygous for cathepsin C mutations inherited from a common ancestor. One patient was heterozygous and showed plantar hyperkeratosis without periodontal disease. Two other family members were also heterozygous but did not present palmoplantar hyperkeratosis and/or periodontal disease. CONCLUSIONS: This study describes a novel mutation of the cathepsin C gene in a Brazilian kindred with Papillon Lefevre syndrome. PMID- 11922262 TI - Periodontal changes in liver cirrhosis and post-transplantation patients. II: radiographic findings:. AB - BACKGROUND: Liver disease and transplantation affect bone turnover. The role of cylosporin A (CsA) in aggravating bone loss is controversial. The aim of the present study was to examine the effect of liver cirrhosis, transplantation, and immunosuppressive therapy with either CsA or tacrolimus on alveolar bone height. METHODS: The experimental group consisted of 13 liver cirrhosis (LC) patients. A second experimental group included 24 post-liver transplantation patients (PT) receiving CsA or tacrolimus. Seventeen healthy subjects formed a control group. Panoramic x-rays were taken and digitized using a computer-based measurement software to assess alveolar bone height of all available teeth. RESULTS: Bone loss in the PT group (4.57+/-0.56 mm) was significantly higher than the control (C) (2.73+/-0.38 mm); however, it was significantly lower (P = 0.0005) than the LC (6.47+/-0.75 mm). Likewise, alveolar bone loss showed a trend for negative correlation (R = 0.404, P = 0.06) with the duration of immunosuppressive therapy post-liver transplantation. CONCLUSIONS: Liver cirrhosis patients demonstrated greater bone loss compared to healthy controls. Restoration of liver functions following transplantation seems to have the potential to reverse some of these radiographic changes. Further longitudinal studies will be necessary to substantiate these findings. PMID- 11922263 TI - The causes of early implant bone loss: myth or science? AB - The success of dental implants is highly dependent on integration between the implant and intraoral hard/soft tissue. Initial breakdown of the implant-tissue interface generally begins at the crestal region in successfully osseointegrated endosteal implants, regardless of surgical approaches (submerged or nonsubmerged). Early crestal bone loss is often observed after the first year of function, followed by minimal bone loss (< or =0.2 mm) annually thereafter. Six plausible etiologic factors are hypothesized, including surgical trauma, occlusal overload, peri-implantitis, microgap, biologic width, and implant crest module. It is the purpose of this article to review and discuss each factor Based upon currently available literature, the reformation of biologic width around dental implants, microgap if placed at or below the bone crest, occlusal overload, and implant crest module may be the most likely causes of early implant bone loss. Furthermore, it is important to note that other contributing factors, such as surgical trauma and penimplantitis, may also play a role in the process of early implant bone loss. Future randomized, well-controlled clinical trials comparing the effect of each plausible factor are needed to clarify the causes of early implant bone loss. PMID- 11922264 TI - External root resorption following partial-thickness connective tissue graft placement: a case report. AB - We report an unusual case of external root resorption (ERR) that developed in a 37-year-old black male approximately 1 year following routine partial thickness connective tissue graft surgery. The lesion was accessed via flap surgery, thoroughly root planed, and the mucoperiosteal flap replaced. The site healed uneventfully and the patient has been closely observed for over 1 year without symptoms or recurrence of the resorptive lesion and the affected tooth remained vital. Clinicians performing partial-thickness connective tissue grafts should be alert to the possible occurrence of root resorption over extended periods of time. The authors speculate that retention of the donor periosteum with placement on the recipient dentin and root biomodification may limit the resorptive response following connective tissue graft procedures to treat tooth root recession. PMID- 11922265 TI - Formation of a cyst-like area after a connective tissue graft for root coverage. AB - BACKGROUND: Many techniques have been developed to obtain root coverage. There have been few complications reported with these techniques. This case report chronicles the development of a cyst-like area after a connective tissue partial thickness double pedicle graft was used to obtain root coverage. METHODS: The root coverage procedure was performed as previously reported. Postoperatively, in the surgical area, the patient developed occasional swelling. It was possible to express a white material from the area. A biopsy was taken to determine the cause of the clinical findings. RESULTS: Histological evaluation revealed the development of a cyst-like area. CONCLUSION: The development of cysts does not seem to be a common event. However, this case report demonstrates that they are a possible complication of root coverage procedures. PMID- 11922266 TI - Application of enamel matrix derivative in autotransplantation of an impacted maxillary premolar: a case report. AB - BACKGROUND: The success of tooth transplantation or replantation depends on the viability of periodontal ligament in the planted tooth. Mechanical injury to periodontal tissues frequently results in dental root resorption and dental ankylosis, which leads to the failure of transplantation or replantation. Enamel matrix derivative (EMD) has been recently used to induce periodontal regeneration. In this report, we show a clinical case of EMD application in the transplantation of an inversely impacted and immature tooth. METHODS: An impacted second premolar was found in the right maxilla of a 16 year-old girl. The tooth was inversely impacted and the dental root was incomplete. When transplantation was carried out, EMD was applied to the periodontal tissues of the extracted premolar. The tooth was fixed at the correct position and the clinical condition was followed for evaluation for 6 months. RESULTS: Radiographs after 3 months exhibited new bone formation surrounding the transplanted tooth. After 6 months, considerable growth of dental root was evident, periodontal ligament-like radiolucency appeared, the vital reaction of the planted tooth was detected, and there were no signs of root resorption or ankylosis. CONCLUSION: Short-term results from this case indicate that EMD application was effective in the transplantation of an inversely impacted and immature tooth and that EMD might contribute to the growth of dental root and to the prevention of ankylosis. PMID- 11922267 TI - Dilated cardiomyopathy: concepts derived from gene deficient and transgenic animal models. AB - Genetic forms of human dilated cardiomyopathy (DCM) are briefly discussed, and a variety of animal models of genetic DCM are presented, some of which are caused by the gene mutations that also cause DCM in humans. The forms of DCM related to mutations or deletion of genes coding for extrasarcomeric or intrasarcomeric proteins, as well as to overexpression or knockout of genes in the beta adrenergic signaling pathway, are included. Finally, novel approaches to treatment in experimental animal models are discussed, including double transgenesis and newer recombination methods, as well as in vivo somatic gene transfer which, based on initial experiments in animals, seems likely to find eventual application in human cardiac failure. PMID- 11922268 TI - Proliferative signaling and disease progression in heart failure. AB - Therapy for heart failure has traditionally been directed to such short-term functional abnormalities as low cardiac output, high filling pressures, and fluid retention. More recently, it has become clear that therapy must also inhibit the proliferative responses that contribute to the progressive deterioration of the failing heart. That heart failure is more than a hemodynamic disorder became apparent when clinical trials showed that drugs that improve such functional abnormalities as high venous pressure and low cardiac output failed to improve long-term prognosis. Most vasodilators, in spite of alleviating short-term problems caused by excessive afterload, increase long-term mortality; the notable exceptions are ACE inhibitors, the ability of which to prolong survival and inhibit remodeling can be attributed to inhibition of proliferative signaling. Other clinical trials showed that inotropic drugs, while providing immediate relief of symptoms, generally shorten long-term survival, whereas beta-adrenergic receptor blockers, which inhibit proliferative signaling by norepinephrine, improve prognosis. These findings can be explained by crossovers between functional and proliferative signaling, among the most important of which is the ability of neurohumoral mediators, such as norepinephrine and angiotensin II, to stimulate maladaptive hypertrophy, remodeling, apoptosis and other deleterious proliferative responses in the failing heart. The emerging understanding of the role of cytoskeletal and cell adhesion molecules in activating maladaptive proliferative responses suggests additional targets for therapy, and the rapid pace of discovery in molecular biology promises additional opportunities to inhibit this abnormal signaling, which causes progressive ventricular dilatation (remodeling) and cardiac cell death, now recognized to be major problems in this syndrome. PMID- 11922269 TI - Triggers and circadian distribution of the onset of acute aortic dissection. AB - The purpose of this study was to clarify the activities that trigger the onset of acute aortic dissection (AAD) and their relation to the occurrence of AAD. The study group comprised 444 consecutive patients referred for spontaneous AAD. From the hospital medical records, the activities that triggered AAD could be identified in 307 of these: most (86.6%) AAD episodes occurred in relation to physical (73.6%) or mental (13%) activities. In the older (> or =61 years) population, AAD occurred significantly more frequent during sleep or rest than in the younger (< or =60 years) population (16.9% vs 7.6%, p=0.020). The relationship between the time of onset and the triggering activity of AAD could be assessed in 267 patients. The onset of AAD was predominantly during the day: 63.3% of the episodes occurred between 06.00h and 18.00h, and were significantly more related to physical or mental activities than the nighttime events (95.3% vs 70.4%, p<0.0001). Most (86.6%) of the AAD episodes were related to physical or mental stress, particularly those that occurred during the day. PMID- 11922270 TI - Atrial components contributing to pseudo r' deflection in lead V1 in slow/fast atrioventricular nodal reentrant tachycardia: analysis of the atrial activation sequence by basket catheter isochronal mapping. AB - Electrocardiographic recognition of the P' wave during tachycardia is very useful in the diagnosis of supraventricular tachycardias. In slow/fast (S/F) atrioventricular nodal reentrant tachycardia (AVNRT), no discrete P' waves are observed on ECG and pseudo r' deflection in lead V1 (pseudo r') is commonly recognized. However, the atrial components that contribute to the genesis of pseudo r' in lead V1 have not been described and this study aimed to clarify them by analysis of the whole activation sequence of the right atrium using Basket catheter isochronal mapping. The study group comprised 48 patients with AVNRT. Pseudo r' was defined as an upward deflection in the terminal portion of the QRS complex during tachycardia that was not recognized during sinus rhythm and it occurred in 45 patients (94%). During S/F AVNRT, the retrograde atrial activation was earliest on His bundle electrogram, followed by the coronary sinus ostium, distal coronary sinus and high right atrium. Only the high lateral aspect of the right atrium was activated after the end of the QRS complex. The interval between the onset of QRS in multiple surface ECG leads and the atrial activities on high right atrium was similar to the V-r' interval in lead V1 (111+/-20ms, 117+/-11 ms) and correlated with the V-r' interval (r=0.56). Pseudo r' deflection in lead V1 is a highly sensitive indicator of S/F AVNRT, and appears to result from the activation of the superolateral aspect of the right atrium. PMID- 11922271 TI - Effects of a single oral dose of cilostazol on epicardial coronary arteries and hemodynamics in humans. AB - Cilostazol, a novel cyclic adenosine monophosphate phosphodiesterase type III inhibitor, has been developed as an antiplatelet drug with a vasodilating action on peripheral arteries. The present study was designed to test, in humans, whether cilostazol can dilate the epicardial coronary arteries and what are its hemodynamic effects. Eight patients with chest pain syndrome were subjected to serial quantitative coronary arteriography immediately before and at 30, 60 and 150min after a single oral dose of cilostazol (200mg). Luminal cross-sectional areas (mm2) at the proximal and distal sites of major coronary arteries (6 segments at each sampling time) were significantly increased at 150 min after taking the drug. The percent increases relative to the baseline values were 25+/ 7 (6.8+/-0.8-->8.3+/-1.0*) and 42+/-7% (2.1+/-0.3-->3.0+/-0.4*) in the right coronary artery, 24+/-5 (5.1+/-0.7-->6.1+/-0.8*) and 28+/-10% (1.6+/-0.31-->9+/ 0.3*) in the left anterior descending artery, and 14+/-6 (5.9+/-0.9-->6.6+/-0.9*) and 24+/-10% (1.3+/-0.2-->1.5+/-0.2*) in the left circumflex artery, respectively (*p<0.05 vs baseline). This action, relative to that of nitroglycerine, was between 27% and 54%. Moreover, small but sustained decreases in systolic pulmonary pressure and stroke work index were observed. Thus, cilostazol has a mild coronary vasodilating action with minimal hemodynamic effects, thereby giving it a possible role in the treatment of coronary artery disease. PMID- 11922272 TI - Relationship between cardiac sympathetic function and baroreceptor sensitivity after acute myocardial infarction. AB - The relationship between baroreceptor sensitivity (BRS) and cardiac sympathetic nerve function after acute myocardial infarction (AMI) was investigated in 34 patients. BRS was measured during the Valsalva maneuver and cardiac sympathetic function was assessed by the washout rate (WR) of I-123 meta-iodo-benzyl guanidine (123I-MIBG) with planar image (global WR) and polar map analysis (regional WR). Gated left ventricular ejection fraction (LVEF) measured by left ventriculography as a parameter of ventricular function was measured with quantitative gated SPECT (QGS). The gated LVEF correlated with global WR (r= 0.36, p=0.034) and regional WR of normal area (r=-0.46, p=0.006), but not with BRS, although BRS correlated with global WR (r=-0.43, p=0.015) and regional WR of normal area (r=-0.72, p<0.0001). After AMI, baroreceptor function is linked to sympathetic activation, as elucidated by the regional WR of normal area, which suggests that separation of the infarcted area from the non-infarcted myocardium is necessary for evaluating sympathetic activation after AMI and that the regional kinetics of 123I-MIBG in the normal area are a more suitable marker of activated cardiac nerve function than global 123I-MIBG kinetics. PMID- 11922273 TI - Hepatocyte growth factor production may be related to the inflammatory response in patients with acute myocardial infarction. AB - Hepatocyte growth factor (HGF) is a well-known powerful proliferative factor of vascular endothelial cells and it has been reported that plasma HGF concentrations are increased in acute myocardial infarction (AMI), although the mechanisms are not yet well delineated. Serum HGF levels and C-reactive protein (CRP) were measured in 22 patients with unstable angina pectoris (UAP) (15 males, 7 females; class IIb or IIIb of the Braunwald classification), 60 patients with AMI (37 males, 23 females; average time from the onset of symptoms to admission 4.6+/-0.7h, range, 0.5-12h), and 20 normal subjects. Immediate angioplasties were performed in 51 patients with AMI, and the time course of the HGF levels were measured in 31 patients among them. Heparin dramatically increased the HGF level and it declined to the normal range 18h after heparin injection. Blood samples were taken before heparin treatment, or at least 24h after. Serum HGF levels on admission was significantly increased in UAP (mean+/-SE: 0.30+/-0.03ng/ml, p<0.01), and AMI (0.27+/-0.02ng/ml, p<0.01) compared with the normal subjects (0.19+/-0.01 ng/ml). Even in the early stage (within 3 h of onset of symptoms to admission, average time was 1.8+/-0.1 h), serum HGF levels were already elevated (0.25+/-0.02 ng/ml, p<0.05). There was no significant difference between the HGF levels in UAP and AMI. Fifty-one of the 60 patients with AMI underwent immediate percutaneous transluminal coronary angioplasty and blood samples were obtained from 31 of them on days 7, 14, and 21 after MI. Serum HGF levels peaked on day 7 (0.34+/-0.04ng/ml, p<0.01) and there was a weak relationship between peak creatine kinase and serum HGF levels at that time. A statistically significant correlation was found between peak CRP and serum HGF levels on day 7 (r=0.62: p<0.001). Serum HGF levels decreased to nearly normal by day 21 (0.22+/-0.01 ng/ml). The study shows that serum HGF levels during the early stage of AMI increased significantly and peaked by day 7 after the onset, at which time there was a strong correlation with peak CRP levels. These data suggest that HGF production may be related to the inflammatory response in AMI. PMID- 11922274 TI - Incidence and clinical characteristics of chronic pulmonary thromboembolism in Japan compared with acute pulmonary thromboembolism: results of a multicenter registry of the Japanese Society of Pulmonary Embolism Research. AB - The incidence of acute pulmonary thromboembolism (APTE) in Japan is quoted as being extremely low compared with the United States, and the incidence and clinical characteristics of chronic pulmonary thromboembolism (CPTE) in Japan is unknown, so this study investigated these aspects of CPTE in 309 patients with APTE and 68 patients with CPTE. The ratio of the incidence of CPTE to APTE was 0.22 and there was no significant difference in age or sex between the APTE and CPTE patients. All of the predisposing factors for pulmonary thromboembolism, except for thrombophilia, were more frequently seen in the patients with APTE. There are some differences in the incidence and clinical characteristics of CPTE compared with APTE between Japanese and American patients in Japan, suggesting that the pathogenesis of CPTE in Japan may differ from that in the USA. PMID- 11922275 TI - Prognostic significance of exercise plasma noradrenaline levels for cardiac death in patients with mild heart failure. AB - The present study was designed to determine whether exercise plasma noradrenaline (NA) levels could predict cardiac death in patients with mild heart failure in whom the plasma NA levels were only minimally elevated. Treadmill exercise testing with serial measurement of plasma NA and plasma adrenaline were performed in 142 patients with heart failure (New York Heart Association class I-II; age, 58+/-12 years) and 26 age-matched normal subjects. During a median follow-up of 9.6 years, 27 cardiac deaths occurred among the patients. By univariate Cox proportional hazard analysis, left ventricular end-systolic dimension (p<0.001), age (p<0.01), peak exercise heart rate (p<0.01), exercise plasma NA level (p<0.01) and left ventricular ejection fraction (p<0.001) were identified as significant prognostic markers. In a multivariate analysis, exercise plasma NA level was identified as the most powerful prognostic marker (p<0.001), followed by left ventricular end-systolic dimension and peak exercise heart rate. In addition, from the Kaplan-Meier analysis, patients with a supramedian level of exercise plasma NA concentration (NA > or =840pg/ml) had a significantly lower survival rate than those with an inframedian level (p<0.01). Exercise plasma NA levels can provide prognostic information in patients with mild heart failure, which suggests an important role of exercise-induced activation of sympathetic nervous system activity in the prognosis of patients with mild heart failure. PMID- 11922276 TI - Association between plasma lipoprotein(a) and endothelial dysfunction in normocholesterolemic and non-diabetic patients with angiographically normal coronary arteries. AB - The present study was designed to examine whether elevated levels of lipoprotein(a) (Lp(a)) are related to the impairment of the endothelium-dependent vasoresponse to acetylcholine (ACh) in normocholesterolemic and non-diabetic human normal coronary arteries. ACh (30 microg) was injected into the left main coronary artery of 31 patients (serum low-density cholesterol <160mg/dl and fasting plasma glucose <126mg/dl) with angiographically normal coronary arteries, and the relation between diameter change and lipid levels was analyzed. The mean diameter change of all coronary segments examined (segments 6, 8, 11 and 13) was reduced by 14.6+/-26.5% in response to ACh, but increased by 23.3+/-6.0% in response to nitroglycerin, suggesting endothelial dysfunction in those arteries. The mean diameter change of the left anterior descending artery or left circumflex artery in each patient was negatively correlated only with the level of Lp(a). Stepwise multiple regression analysis also revealed that only Lp(a) among the lipids showed significant correlation with impaired vasodilation (p=0.033). These findings suggest that elevated levels of plasma Lp(a) might be a strong predictor of endothelial dysfunction in normocholesterolemic and non diabetic subjects. PMID- 11922277 TI - In vivo echocardiographic detection of cardiovascular lesions in apolipoprotein E knockout mice using a novel high-frequency high-speed echocardiography technique. AB - Apolipoprotein E-knockout (apoE-KO) mice have been used for studying atherogenesis, but the in vivo features including cardiovascular function have not yet been reported. This study aimed to noninvasively evaluate cardiovascular lesions in 6 apoE-KO mice and 6 control (C57BL/6) mice using transthoracic echocardiography performed using an originally developed linear scanner that permits a high-speed scan with wideband high-frequency ultrasound. Two independent observers evaluated and scored the degree of atherosclerotic changes in the aortic root from 2-dimensional long-axis and short-axis images. M-mode measurements included left ventricular end-diastolic dimension (LVDd), posterior wall thickness (LVPWT), fractional shortening, aortic root dimension and rate of systolic expansion of the aorta (%SEAo). The wall thickness of the aortic root was measured from the serial histological sections. Significant differences between apoE-KO and C57BL/6 mice were found in the atherosclerotic score, %SEAo, LVDd and LVPWT. The atherosclerotic score and %SEAo were significantly correlated with the aortic wall thickness. Transthoracic echocardiography with a high frequency ultrasound system can detect atherosclerotic lesions and the decreased distensibility of the ascending aorta, as well as secondary changes in left ventricular geometry, in apoE-KO mice. PMID- 11922278 TI - Vasopressin inhibits sarcolemmal ATP-sensitive K+ channels via V1 receptors activation in the guinea pig heart. AB - To examine the effect of vasopressin on the sarcolemmal ATP-sensitive K (K(ATP)) channel, cell-attached, insideout and open-cell-attached methods of patch clamp techniques were used in isolated guinea pig ventricular myocytes. Suppressing both glycolytic and oxidative ATP production attained K(ATP) channel activation. In the cell-attached mode, vasopressin inhibited KATP channels in a concentration dependent manner with an IC50 of 15.1+/-1.8 nmol/L. In the inside-out configuration, vasopressin failed to block K(ATP) channels. In the cell-attached mode, manning compound (1 micromol/L), a V1 receptor-selective antagonist, blocked the inhibitory action of vasopressin, although OPC-31260 (1 micromol/L), a V2 receptor-selective antagonist could not affect the action of vasopressin. In addition, vasopressin lost its inhibitory action on K(ATP) channels when the channel was activated by pinacidil, a K channel opener and in the open-cell attached mode effected by streptolysin-O. Thus, the inhibitory action of vasopressin K(ATP) channels may occur via V1 receptor related mechanism. PMID- 11922279 TI - Inhomogeneous derangement of cardiac autonomic nerve control in diabetic rats. AB - The present study compared autonomic nervous function in Kob [Spontaneously Diabetic, Bio-Breeding (BB)] rats with control Wistar rats to determine the development of cardiac neuropathy in diabetic rats. Telemetric ECG signals were obtained from an ECG radio-transmitter placed in a dorsal subcutaneous pouch of male Kob and Wistar rats for 30min every 6h at a sample rate of 5kHz. Heart rate (HR) and HR variability (HRV) were analyzed in each group by power spectrograms obtained by a fast Fourier transform algorithm. RR interval, total power (TP), low frequency (LF) power (0.04-0.67 Hz), high frequency (HF) power (0.79-1.48 Hz) and LF/HF ratio were also measured. The Kob rats had lower HRV than the control Wistar rats; HR, TP, and HF power, but not the LF/HF ratio, in the Kob rats were significantly lower than those of the control rats (p<0.001). However, in the Kob rats the response of these parameters to a muscarinic antagonist (atropine: 2mg/kg) was left intact, but their response to a beta-adrenergic antagonist (propranolol: 4mg/kg) was impeded. Autonomic nervous control of HR in spontaneously diabetic rats was inhomogeneously deranged in terms of the balance in sympathetic and parasympathetic tone, not only in the baseline condition, but also in the regulatory systems, including postsynaptic receptor function. PMID- 11922281 TI - Recovery of cardiac function after living-donor lung transplantation in a patient with primary pulmonary hypertension. AB - Legislation for brain death and organ transplantation was passed in Japan in 1997, but there is still a great shortage of brain-death donors. Primary pulmonary hypertension (PPH) is a progressive disease that is usually followed by death within 5 years of diagnosis. Continuous infusion of prostacyclin is effective, but some patients will ultimately require heart-lung or lung transplantation. The first case of bilateral living-donor lobar lung transplantation (LDLLT) for PPH in Japan is reported. The recipient was a 19-year old woman who was diagnosed as PPH at the age of 14 years and began intravenous prostacyclin therapy. Initially her symptoms improved, but she returned to New York Heart Association class IV in 2000. In January 2001, she underwent bilateral LDLLT. Postoperative echocardiography showed that the right ventricular diameter had decreased and septal wall motion had normalized, resulting in a round-shaped left ventricle. Right heart catheterization demonstrated that cardiac output and pulmonary arterial pressure had normalized. The right ventricular ejection fraction improved from 15% to 77%. The patient was discharged from hospital after 60 days postoperatively. LDLLT will become one of the options in Japan for end stage PPH. PMID- 11922280 TI - Nipradilol can prevent left ventricular systolic and diastolic dysfunction after myocardial infarction in rats. AB - The purpose of this study was to examine the effects of nipradilol on the cardiac function and mRNA expression in Wistar rats with a myocardial infarction (MI) that was created by ligation of the anterior descending coronary. Ten mg x kg(-1) x day(-1) of nipradilol were administrated to the rats in random order, and hemodynamic and Doppler-echocardiographic findings and myocardial mRNA expression were analyzed at 4 weeks after MI. Although left ventricular end-diastolic pressure (LVEDP) and central venous pressure (CVP) were increased in the MI rats, nipradilol significantly reduced the degree of the increase in both parameters. MI also significantly increased the weight of the left and right ventricles, and increased the left ventricular end-diastolic dimension (LVDd), effects that were attenuated by nipradilol. The MI rats showed decreased fractional shortening as systolic dysfunction and decreased E wave deceleration rate as diastolic dysfunction, and nipradilol significantly prevented these. Nipradilol significantly suppressed the increase in the non-infarcted myocardial mRNA expression of atrial natriuretic peptide, brain natriuretic peptide and collagen I and III. In conclusions, nipradilol prevents the cardiac remodeling that is accompanied by systolic and diastolic dysfunction, and inhibits abnormal myocardial gene expression after MI. PMID- 11922282 TI - Hyponatremic-hypertensive syndrome associated with renovascular hypertension: a case report. AB - Renovascular hypertension occasionally manifests clinically as electrolyte disorders and albuminuria in addition to elevated blood pressure. A 49-year-old man who had renovascular hypertension also had severe hypokalemia, hyponatremia, polyuria and polydipsia that were treated by an angiotensin-converting enzyme inhibitor and resection of an atrophic kidney with a compromised blood supply. This is a case of hyponatremic-hypertensive syndrome related to renovascular hypertension and occurring as an additional abnormality. PMID- 11922283 TI - Percutaneous transseptal mitral valvuloplasty in the presence of undegenerated septum primum. AB - A 37-year-old woman had progressive shortness of breath and mitral stenosis was diagnosed. Despite the unusual finding of undegenerated septum primum on echocardiography and angiography, percutaneous transseptal mitral commissurotomy was successfully performed in this patient with rheumatic mitral stenosis under the guidance of online transesophageal echocardiography. PMID- 11922284 TI - Infected papillary fibroelastoma attached to the atrial septum. AB - A 61-year-old woman had intermittent fever of 2 months' duration following a dental extraction. On admission, her body temperature was 39.2 degrees C. A mid systolic murmur was heard at the apex on ausculation. A 2-dimensional echocardiogram revealed a mobile, heavy stick-like mass with vegetation (5.0 x 1.5 cm) attached to the left atrial septum. Multiple blood cultures grew Streptococcus constellatus. On diagnosis of an infected left atrial myxoma, antibiotics were administered daily and 4 weeks later, the left atrial tumor was resected. The tumor was 5.3cm long, 1.5cm in diameter at the inter-atrial wall and had vegetation on the free edge. On microscopic examination, colonies of Gram positive cocci were found in the thrombus, on the papillary fibroelastoma. After treatment with antibiotics for a further 4 weeks, the patient was discharged. This is the first report of infected papillary fibroelastoma. PMID- 11922285 TI - Action potential duration, activation-recovery interval and effective refractoriness: a well known but unresolved trinity. PMID- 11922286 TI - A microfabricated thermal field-flow fractionation system. AB - A microscale thermal field-flow fractionation (micro-TFFF) system has been designed, fabricated, and characterized. Motivation for miniaturization of TFFF systems was established by examining the geometrical scaling of the fundamental TFFF theory. Miniaturization of conventional macroscale TFFF systems was made possible through utilization of micromachining technologies. Fabrication of the micro-TFFF system was discussed in detail. The micro-TFFF system was characterized for plate height versus flow rate, single-component polystyrene retention, and multicomponent polystyrene separations. Retention, thermal diffusion coefficients, and maximum diameter-based selectivity values were extracted from separation data and found comparable with macroscale TFFF system results. Retention values ranged from 0.33 to 0.46. Thermal diffusion coefficients were between 3.0 x 10(-8) and 5.4 x 10(-8) cm2/s x K. The maximum diameter-based selectivity was 1.40. PMID- 11922287 TI - Separation in slalom chromatography: stretching and velocity dependence. AB - Slalom chromatography (SC) is used for the separation of large double-stranded DNA molecules. In this technique, the progression of the DNA fragments through the closed column packing follows the flow direction and is like a snake edging is way into long grass. A novel mathematical model is developed in this paper to describe this hydrodynamic phenomenon. The results obtained provided a model for the resolution between two adjacent peaks on a chromatogram. As well, a chromatographic response function was used to obtain the most efficient separation conditions for a mixture of DNA fragments with sizes higher than 15 kbp in a minimum analysis time. PMID- 11922288 TI - A high-throughput continuous sample introduction interface for microfluidic chip based capillary electrophoresis systems. AB - The development of efficient sample introduction and pretreatment systems for microfluidic chip-based analytical systems is important for their application to real-life samples. In this work, world-to-chip interfacing was achieved by a novel flow-through sampling reservoir featuring a guided overflow design. The flow-through reservoir was fabricated on a 30 x 60 x 3 mm planar glass chip of crossed-channel design used for capillary electrophoresis separations. The 20 microL sample reservoir was produced from a section of plastic pipet tip and fixed at one end of the sampling channel. Sample change was performed by pumping 80-microL samples sandwiched between air segments at approximately 0.48 mL/min flow rate through the flow-through reservoir, introduced from an access hole on the bottom side of the chip. A filter paper collar wrapped tightly around the reservoir guided the overflowing sample solution into a plastic trough surrounding the reservoir and then to waste. The performance of the system was demonstrated in the separation and determination of FITC-labeled arginine, glycine, phenylalanine, and glutamic acid with LIF detection, by continuously introducing a train of different samples through the system without electrical interruption. Employing a separation channel of 4 cm (2-cm effective separation length) and 1.4-kV separation voltage, maximum throughputs of 80/h were achieved with <4.1% carryover and precisions ranging from 1.5% for arginine to 2.6% RSD (n = 11) for glycine. The sampling system was tested in the continuous monitoring of the derivatizing process of amino acids by FITC over a period of 4 h, involving 166 analytical cycles. An outstanding overall precision of 4.8% RSD (n = 166) was achieved for the fluorescein internal standard. PMID- 11922289 TI - Feasibility of collection and analysis of airborne carbonyls by on-sorbent derivatization and thermal desorption. AB - The most commonly used method for analysis of airborne carbonyls is to collect the analytes on solid sorbents coated with a suitable derivatization agent, followed by solvent desorption and liquid injection for analysis by high-pressure liquid chromatography. We have explored a new approach by combining on-sorbent derivatization and thermal desorption to measure airborne carbonyls. More specifically, carbonyls in the air are collected onto an O-(2,3,4,5,6 pentafluorobenzyl)hydroxylamine (PFBHA)-coated Tenax sorbent packed in a tube with dimensions identical to those of a gas chromatography (GC) injector liner. The derivatives are then released by in-injection port thermal desorption to a GC column for analysis. Gaseous carbonyls, including formaldehyde, acetaldehyde, benzaldehyde, glyoxal, and methylglyoxal, at ppbv levels are shown to be effectively collected (> or = 92% collection efficiency) onto the sampling tubes at a flow rate of 20 mL/min under both < 1% and 71% relative humidity. The collection efficiency drops as the sampling flow rate increases, and the degree of decrease is compound dependent. The derivatization agent, at a level of approximately 127 nmol/tube, is thermally desorbed and eluted from the GC column without compromising the determination of any carbonyl-PFBHA derivatives. Detection limits of low ppbv to sub-ppbv are achieved for a sample air volume of 4.8 L. Using this new method, we have measured formaldehyde, acetaldehyde, benzaldehyde, glyoxal, and methylglyoxal to be 4.9-16.3, 0.6-8.2, <5.9, 0.5-4.1, and <2.4 ppbv, respectively, in the ambient atmosphere at the university bus stop. This method is less labor intensive than the solvent desorption technique and avoids use of organic solvents. Other classes of airborne polar species can be measured through the same approach by selecting an appropriate derivatization agent. PMID- 11922290 TI - Electrokinetic chromatography utilizing two pseudostationary phases providing ion exchange and hydrophobic interactions. AB - The electrokinetic chromatographic separation of a series of inorganic and organic anions was achieved by utilizing an electrolyte system comprising a cationic soluble polymer (poly(diallydimethylammonium chloride, PDDAC) and a neutral beta-cyclodextrin (beta-CD) as pseudostationary phases. The separation mechanism was a combination of electrophoresis, ion-exchange (IE) interactions with PDDAC, and hydrophobic interactions with beta-CD. The extent of each chromatographic interaction was independently variable, allowing for control of the separation selectivity of the system. IE interactions could be varied by changing either the PDDAC concentration or the concentration of a competing ion (e.g., chloride) in the BGE, while the hydrophobic interactions could be varied by changing the concentration of beta-CD. The separation system was very robust, with the reproducibility of the migration times being <0.7% RSD. A mathematical model that predicted the mobilities of analytes under varying experimental conditions was derived and was shown to give good correlation (r2 = 0.9804) between predicted and experimental migration times. Parameters derived from the model were in good agreement with the ion-exchange and hydrophobic characteristics of the analytes. The model was also applied successfully to the optimization of conditions for the separation of a mixture of analytes or for conditions under which particular analytes migrated in a desired order. That is, the opportunity to tune the separation selectivity has been demonstrated. PMID- 11922291 TI - Examination of theoretical models in external voltage control of capillary electrophoresis. AB - Control of electroosmosis by an external voltage in capillaries of varying geometry was examined and investigated. Controlled geometric factors included inner and outer radii, external electrode coverage area, and the method of voltage application. The behavior of the flow in response to the external voltage from earlier work and this study were compared to existing literature models. A noticeable lack of correlation between the current modeling theories and the published data is noted. In light of these results, suggestions for further areas of investigation of a description of external voltage flow control mechanism are suggested. PMID- 11922292 TI - Field measurement of acid gases and soluble anions in atmospheric particulate matter using a parallel plate wet denuder and an alternating filter-based automated analysis system. AB - We present a new fully automated instrument for the measurement of acid gases and soluble anionic constituents of atmospheric particulate matter. The instrument operates in two independent parallel channels. In one channel, a wet denuder collects soluble acid gases; these are analyzed by anion chromatography (IC). In a second channel, a cyclone removes large particles and the aerosol stream is then processed by another wet denuder to remove potentially interfering gases. The particles are then collected by one of two glass fiber filters which are alternately sampled, washed, and dried. The washings are preconcentrated and analyzed by IC. Detection limits of low to subnanogram per cubic meter concentrations of most gaseous and particulate constituents can be readily attained. The instrument has been extensively field-tested; some field data are presented. Results of attempts to decipher the total anionic constitution of urban ambient aerosol by IC-MS analysis are also presented. PMID- 11922294 TI - Accelerated solvent extraction followed by on-line solid-phase extraction coupled to ion trap LC/MS/MS for analysis of benzalkonium chlorides in sediment samples. AB - Benzalkonium chlorides (BACs) were successfully extracted from sediment samples using a new methodology based on accelerated solvent extraction (ASE) followed by an on-line cleanup step. The BACs were detected by liquid chromatography/ion trap mass spectrometry (LC/MS) or tandem mass spectrometry (MS/MS) using an electrospray interface operated in the positive ion mode. This methodology combines the high efficiency of extraction provided by a pressurized fluid and the high sensitivity offered by the ion trap MS/MS. The effects of solvent type and ASE operational variables, such as temperature and pressure, were evaluated. After optimization, a mixture of acetonitrile/water (6:4 or 7:3) was found to be most efficient for extracting BACs from the sediment samples. Extraction recoveries ranged from 95 to 105% for C12 and C14 homologues, respectively. Total method recoveries from fortified sediment samples, using a cleanup step followed byASE, were 85% for C12BAC and 79% for C14BAC. The methodology developed in this work provides detection limits in the subnanogram per gram range. Concentrations of BAC homologues ranged from 22 to 206 microg/kg in sediment samples from different river sites downstream from wastewater treatment plants. The high affinity of BACs for soil suggests that BACs preferentially concentrate in sediment rather than in water. PMID- 11922293 TI - Chemiluminometric sensor for simultaneous determination of L-glutamate and L lysine with immobilized oxidases in a flow injection system. AB - A chemiluminometric flow-through sensor for simultaneous determination of L glutamate (Glu) and L-lysine (Lys) in a single sample has been developed. Immobilized uricase, immobilized peroxidase, support material, coimmobilized glutamate oxidase/peroxidase, support material, and coimmobilized lysine oxidase/peroxidase were packed sequentially in a transparent PTFE tube, and the tube was placed in front of a photomultiplier tube as a flow cell. A three-peak recording was obtained by one injection of the sample solution. The peak height of the first peak was due to the concentrations of urate and other reductants in the sample; the immobilized uricase was used to decompose urate, and the hydrogen peroxide produced was decomposed with a luminol-hydrogen peroxide reaction by immobilized peroxidase. The peak heights of the second and third peaks were free from the interferences from the reductants and were dependent only on the concentrations of Glu and Lys, respectively. Calibration graphs for Glu and Lys were linear at 40-1,000 and 50-1,200 nM, respectively. The sampling rate was 11/h without carryover. The sensor was stable for two weeks. The sensor system was applied to the simultaneous determination of Glu and Lys in serum. PMID- 11922295 TI - Porous graphitic carbon as stationary phase for LC-ICPMS separation of arsenic compounds in water. AB - A new liquid chromatographic separation method was developed for the speciation of the four main arsenic compounds present in water. Arsenite (As(III)), dimethylarsinic acid (DMA), monomethylarsonic acid (MMA) and arsenate (As(V)) were separated on a recently introduced stationary phase: porous graphitic carbon (PGC). The separation was first obtained under formic acid gradient conditions, but an adsorption phenomenon of As(V) on PGC was observed. To overcome this problem, As(V) was backflushed, and an efficient separation of the four solutes was achieved within 10 min. Extremely low detection limits (ranging from 10 to 70 ng x L(-1)) were obtained by coupling LC with an ICPMS. The method was successfully applied to different spiked mineral waters and a naturally arsenic containing freshwater. PMID- 11922296 TI - Hot microelectrodes. AB - Heat generation at disk microelectrodes by a high-amplitude (few volt) and high frequency (0.1-2 GHz) alternating voltage is described. This method allows changing electrode temperature very rapidly and maintaining it well above the boiling point of solution for a very long time without any indication of boiling. The size of the hot zone in solution is determined by the radius of the electrode. There is no obvious limit in regard to the electrode size, so theoretically, by this method, it should be possible to create hot spots that are much smaller than those created with laser beams. That could lead to potential applications in medicine and biology. The heat-generating waveform does not electrically interfere with normal electroanalytical measurements. The noise level at hot microelectrodes is only slightly higher, as compared to normal microelectodes, but diffusion-controlled currents at hot microelectrodes may be up to 7 times higher, and an enhancement of kinetically controlled currents may be even larger. Hot microelectrodes can be used for end-column detection in capillary electrophoresis and for in-line or in vivo analyses. Temperature gradients at hot microelectrodes may exceed 1.5 x 10(5) K/cm, which makes them useful in studies of Soret diffusion and thermoelectric phenomena. PMID- 11922297 TI - Bioimprinting of polymers and sol-gel phases. Selective detection of yeasts with imprinted polymers. AB - Coated quartz crystal microbalances were modified with a surface-imprinting process using whole yeast cells. These molded polymer and sol-gel surfaces show honeycomb-like structures as shown by atomic force microscopy. Reinclusion of cells allows a selective on-line monitoring of these microorganism concentrations in water over 5 orders of magnitude. The sensitivity to cells holds up in growth media up to 21 g/L. Even cell fragments can be detected in flowing conditions. The highly robust polymers on the sensor devices are suitable for biotechnological applications. PMID- 11922298 TI - Properties of vapor detector arrays formed through plasticization of carbon black organic polymer composites. AB - Arrays of vapor detectors have been formed through addition of varying mass fractions of the plasticizer diethylene glycol dibenzoate to carbon black-polymer composites of poly(vinyl acetate) (PVAc) or of poly(N-vinylpyrrolidone). Addition of plasticizer in 5% mass fraction increments produced 20 compositionally different detectors from each polymer composite. Differences in vapor sorption and permeability that effected changes in the dc electrical resistance response of these compositionally different detectors allowed identification and classification of various test analytes using standard chemometric methods. Glass transition temperatures, Tg, were measured using differential scanning calorimetry for plasticized polymers having a mass fraction of 0, 0.10, 0.20, 0.30, 0.40, or 0.50 of plasticizer in the composite. The plasticized PVAc composites with Tg < 25 degrees C showed rapid responses at room temperature to all of the test analyte vapors studied in this work, whereas composites with Tg > 25 degrees C showed response times that were highly dependent on the polymer/analyte combination. These composites showed a discontinuity in the temperature dependence of their resistance, and this discontinuity provided a simple method for determining the Tg of the composite and for determining the temperature or plasticizer mass fraction above which rapid resistance responses could be obtained for all members of the test set of analyte vapors. The plasticization approach provides a method for achieving rapid detector response times as well as for producing a large number of chemically different vapor detectors from a limited number of initial chemical feedstocks. PMID- 11922299 TI - Introduction to the principles of ultramicroheptodes in ring-disk interactions. AB - The use of ring-disk ultramicroelectrodes as tips as a means of extending the methodology of the scanning electrochemical microscope is described. Electrodes consisting of one centered disk with six interconnected disks surrounding it served as the ring-disk electrodes. Basic experiments illustrate the behavior in the feedback mode and the generation collection mode. The dependence on the electrodes' size, both potentials, the tip-sample distance, and the local properties of the underying sample were studied. When approaching an electrode to the surface of a sample, steady-state collection efficiencies between zero and unity were elegantly altered by adjusting the distance between tip and sample. The shielding factor could be varied between 0.25 and 0.82 the same way. Concerning feedback methods, the results presented illustrate the new principle of applying a precisely located external stimulus as a separate electrochemical means of analyzing the sample's response. Obviously, this technique can be extended to irreversible redox mediators, which are not treated here explicitly, but give this concept even greater flexibility. The interpretation of such data can be deduced directly from this contribution. PMID- 11922300 TI - Perbrominated closo-dodecacarborane anion, 1-HCB11Br11-, as an ion exchanger in cation-selective chemical sensors. AB - The 2,3,4,5,6,7,8,9,10,1 1,12-undecabromocarborane anion, 1-HCB11Br11- (UBC-) has been evaluated for its suitability as an ion exchanger in solvent polymeric membrane electrodes and bulk optodes. Experiments comparing the chemical stability of the perhalogenated carborane anion to that of the best lipophilic tetraphenylborate, 3,5-[bis(trifluoromethyl)phenyl]borate (TFPB-), demonstrated that in the presence of 0.2 M acetic acid TFPB- was completely lost within 6 h, while the concentration of UBC- decreased by less than 10% in the same time period. Thin-film bulk optodes containing BME-44 as potassium-selective ionophore, ETH 5294 as chromo-ionophore, and UBC- as ionic sites exhibited a K+ response similar to analogous optodes containing TFPB-, with comparable selectivities over Na+ and Ca2+. Potentiometric measurements evaluating the selectivity behavior of UBC- in both ionophore-free and ionophore-containing electrodes were performed. Ionophore-free PVC membranes containing UBC- as ion exchanger and either DOS or NPOE as plasticizer also demonstrated selectivity similar to TFPB--containing membranes. Sodium-selective membranes containing the ionophore 4-tert-butylcalix[4]arenetetraacetic acid tetraethyl ester (sodium ionophore X) and UBC- as ionic sites showed a Nernstian response for sodium and selectivity comparable to that found in analogous electrodes containing TFPB-. PMID- 11922301 TI - Green electrochemiluminescence from ortho-metalated tris(2 phenylpyridine)iridium(III). AB - The electrochemiluminescence (ECL) of Ir(ppy)3 (ppy = 2-phenylpyridine) is reported in acetonitrile (CH3CN), mixed CH3CN/H20 (50:50 v/v), and aqueous (0.1 M KH2PO4) solutions with tri-n-propylamine as an oxidative-reductive coreactant. ECL efficiencies (phi(ecl), photons emitted per redox event) of 0.00092 in aqueous, 0.0044 in mixed, and 0.33 in CH3CN solutions for Ir(ppy)3 were obtained using Ru(bpy)3(2+) (bpy = 2,2'-bipyridine) as a relative standard (phi(ecl) = 1). Photoluminescence (PL) efficiencies of 0.039, 0.050, and 0.069 were obtained in aqueous, mixed, and acetonitrile solutions, respectively, compared to Ru(bpy)3(2+) (phi(em) = 0.042). The ECL spectra were identical to photoluminescence spectra (lambda(max) approximately equal to 517 nm), indicating formation of the same metal-to-ligand (MLCT) excited states in both ECL and PL. The ECL is linear over several orders of magnitude in mixed and acetonitrile solution with theoretical detection limits (blank plus three times the standard deviation of the noise) of 1.23 nM in CH3CN and 0.23 microM in CH3CN/ H20 (50:50 v/v). PMID- 11922302 TI - Diffusion and concentration of molecular probes in thermoresponsive poly(N isopropylacrylamide) hydrogels: effect of the volume phase transition. AB - Poly(N-isopropylacrylamide), NIPA, thermoresponsive hydrogels with well-defined concentrations of an electroactive probe, 1,1'-ferrocenedimethanol, Fc(MeOH)2, were prepared. The discontinuous reversible volume phase transition of such gels occurs at 32 +/- 1 degrees C and results in a release of approximately 93% of the solution from the polymeric network. Transport of Fc(MeOH)2 in both swollen and collapsed gels was studied using steady-state voltammetry and chronoamperometry at platinum microelectrodes. The diffusion coefficient of Fc(MeOH)2 in collapsed gels was approximately 2 orders of magnitude smaller than that in swollen gels. UV/vis spectroscopic studies showed that for 3.0% NIPA gel, the concentration of Fc(MeOH)2 in the collapsed phase was approximately 6 times higher than that in released solution and 4.5 times higher than in the original swollen gel. PMID- 11922304 TI - A method to accurately determine the extent of solid-phase reactions by monitoring an intermediate in a nondestructive manner. AB - The application of Fourier transform infrared (FT-IR) spectroscopy to the quantitation of effective nucleoside loading on a solid support, via detection of a reaction intermediate, was developed. 4,5-Dicyanoimidazole (DCI) activator is reacted with a known amount of polystyrene-bound phosphoramidite to form a diisopropylamine-DCI salt complex that is detected by FT-IR spectroscopy at a unique wavenumber of 1104 cm(-1). Based on a linear standard curve, the species' absorbance at 1104 cm(-1) is then directly translated into an appropriate concentration. This concentration characterizes the number of active phosphoramidite sites on the solid support. This method consistently determines the loading extent of nucleosides (deoxy-A/T/G/C) on the solid support to the nearest 0.02 mmol/g. The coupling reaction extent of a deoxy-T derivative with the support-bound phosphoramidite compares well with the nucleoside loading, as determined by the FT-IR-based analytical method. The nucleoside loading was further verified by phosphorus elemental analysis of the 3'-phosphate linker concentration. PMID- 11922303 TI - Determination of thiols by capillary electrophoresis with amperometric detection at a coenzyme pyrroloquinoline quinone modified electrode. AB - A chemically modified electrode has been developed as a detector for the sensitive and selective determination of thiol-containing compounds following capillary electrophoresis separation. Electrodes were constructed by entrapment of the coenzyme pyrroloquinoline quinone (PQQ) into a polypyrrole (PPy) matrix on a 245-microm graphite electrode during electropolymerization of pyrrole in the presence of PQQ. PQQ serves as an efficient biocatalyst to mediate the oxidation of thiols at a substantially reduced overpotential relative to an unmodified electrode. Furthermore, this design takes advantage of the pH-dependent reversible electrochemical properties of PQQ, which facilitates optimization of separation and detection conditions. The PQQ/PPy-modified electrode was incorporated as an end-column detector, and a separation of homocysteine, cysteine, N-acetylcysteine, and glutathione was developed. Detection limits for these four thiols were determined to be 11, 23, 104, and 134 nM, respectively, with mass detection limits ranging from 0.29 to 3.48 fmol. The PQQ/PPy electrode was also found to be very reproducible in run-to-run, day-to-day, and electrode to-electrode comparisons. The utility of this electrode was demonstrated for the detection of cysteine in dietary supplements and human urine, resulting in excellent agreement with reported values. PMID- 11922305 TI - Prediction of peptide ion collision cross sections from topological molecular structure and amino acid parameters. AB - Quantitative structure-property relationships (QSPRs) have been developed to predict the ion mobility spectrometry (IMS) collision cross sections of singly protonated lysine-terminated peptides using information derived from topological molecular structure and various amino acid parameters. The primary amino acid sequence alone is sufficient to accurately predict the collision cross section. The models were built using multiple linear regression (MLR) and computational neural networks (CNNs). The best MLR model found contains six descriptors and predicts 94 of 113 peptides (83%) to within 2% of their experimentally determined values. The best CNN model using the same six descriptors predicts 105 of the 113 peptides (93%) to within 2% of their experimentally determined values. The best overall CNN model, using a different set of six descriptors, predicts 109 of the 113 peptides (96%) to within 2% of their experimentally determined values. In addition, this model can discriminate among peptides having identical amino acid composition, but differing in primary amino acid sequence. This represents a capability not found in previously described models. The descriptors used in the models presented may provide some insight into the nature of peptide ion folding in the gas phase. PMID- 11922306 TI - Combined use of conventional and second-derivative data in the SIMPLISMA self modeling mixture analysis approach. AB - Simple-to-use interactive self-modeling mixture analysis (SIMPLISMA) is a successful pure variable approach to resolve spectral mixture data. A pure variable (e.g., wavenumber, frequency number, etc.) is defined as a variable that has significant contributions from only one of the pure components in the mixture data set. For spectral data with highly overlapping pure components or significant baselines, the pure variable approach has limitations; however, in this case, second-derivative spectra can be used. In some spectroscopies, very wide peaks of components of interest are overlapping with narrow peaks of interest. In these cases, the use of conventional data in SIMPLISMA will not result in proper pure variables. The use of second-derivative data will not be successful, since the wide peaks are lost. This paper describes a new SIMPLISMA approach in which both the conventional spectra (for pure variables of wide peaks) and second-derivative spectra (for pure variables of narrow peaks, overlapping with the wide peaks) are used. This new approach is able to properly resolve spectra with wide and narrow peaks and minimizes baseline problems by resolving them as separate components. Examples will be given of NMR spectra of surfactants and Raman imaging data of dust particle samples taken from a lead and zinc factory's ore stocks that were stored outdoors. PMID- 11922307 TI - Programmable delivery of DNA through a nanopipet. AB - We report the pulsed delivery of single-stranded DNA molecules through a nanopipet. The conical geometry of the pipet leads to a localized electric field, since all of the potential drop occurs in the tip region. Pulsatile delivery of DNA molecules can be achieved in an experimentally simple way with high precision by controlling the applied voltage. Single-molecule detection and fluorescence correlation spectroscopy in the nanopipet enable us to determine the number of molecules delivered. Anomalous slow diffusion of the DNA molecules in the pipet has also been observed. This nanopumping technique may have potential applications in local drug delivery and nanofabrication of biomolecules on surfaces in aqueous environments. PMID- 11922308 TI - Fast characterization of cheeses by dynamic headspace-mass spectrometry. AB - This study describes a rapid method to characterize cheeses by analysis of their volatile fraction using dynamic headspace-mass spectrometry. Major factors governing the extraction and concentration of the volatile components were first studied. These components were extracted from the headspace of the cheeses in a stream of helium and concentrated on a Tenax TA trap. They were then desorbed by heating and injected directly into the source of a mass spectrometer via a short deactivated silica transfer line. The mass spectra of the mixture of volatile components were considered as fingerprints of the analyzed substances. Forward stepwise factorial discriminant analysis afforded a limited number of characteristic mass fragments that allowed a good classification of the batches of cheeses studied. PMID- 11922309 TI - Quantitative analysis of amoxycillin and its major metabolites in animal tissues by liquid chromatography combined with electrospray ionization tandem mass spectrometry. AB - A sensitive and specific method for the quantitative determination of amoxycillin and its major metabolites (amoxycilloic acid, amoxycillinpiperazine-2',5'-dione) in animal tissue samples using liquid chromatography combined with positive electrospray ionization tandem mass spectrometry (LC/ESI-MS/MS) is presented. A liquid extraction using an aqueous 0.01 M potassium dihydrogenphosphate solution as the extraction solvent was performed for a preliminary sample cleanup. The extracts were further purified by a solid-phase extraction using an octadecyl (C18) column. Ampicillin was used as the internal standard. Chromatographic separation of the analytes of interest was achieved on a reversed-phase Hypersil column (100 x 3 mm i.d., dp, 5 microm), using a mixture of 9.6 mM pentafluoropropionic acid in water and acetonitrile as the mobile phase. Gradient elution was performed. To obtain as high sensitivity and selectivity as possible, the mass spectrometer was operated in the multiple reaction monitoring mode. The method was validated for the analysis of amoxycillin and its investigated metabolites in various porcine tissues, kidney, liver, muscle, and fat, according to the requirements defined by the European Community. Calibration graphs were prepared for all tissues, and good linearity was achieved over the concentration range tested (25-500 ng/g, r > or = 0.9974, and goodness of fit < or = 9.6). A limit of quantification of 25 ng/g was obtained for amoxycillin and its metabolites in all tissues, which corresponds to half the maximum residue limit for amoxycillin. Limits of detection ranged from 2.3 to 12.0 ng/g for amoxycillin and from 1.1 to 15.1 ng/g and 0.2 to 2.4 ng/g for amoxycilloic acid and amoxycillinpiperazine-2',5'-dione, respectively. The results for the within-day precision and the trueness fell within the ranges specified. The method has been successfully used for the quantitative determination of amoxycillin and its major metabolites in tissue samples from pigs medicated via the drinking water, proving the usefulness of the developed method for application in the field of residue analysis. PMID- 11922310 TI - A tandem mass spectrometer for improved transmission and analysis of large macromolecular assemblies. AB - We report the design and first applications of a tandem mass spectrometer (a quadrupole time-of-flight mass spectrometer) optimized for the transmission and analysis of large macromolecular assemblies. Careful control of the pressure gradient in the different pumping stages of the instrument has been found to be essential for the detection of macromolecular particles. Such assemblies are, however, difficult to analyze by tandem-MS approaches, because they give rise to signals above m/z 3,000-4,000, the limit for commercial quadrupoles. By reducing the frequency of the quadrupole to 300 kHz and using it as a narrow-band mass filter, we show that it is possible to isolate ions from a single peak at m/z 22,000 in a window as narrow as 22 m/z units. Using cesium iodide cluster signals, we show that the mass range in the time-of-flight (TOF) analyzer extends beyond m/z 90,000, in theory to more than m/z 150,000. We also demonstrate that the resolution of the instrument is greater than 3,000 at m/z 44,500. Tandem-MS capabilities are illustrated by separating components from heterooligomeric assemblies formed between tetrameric transthyretin, thyroxine, retinol-binding protein, and retinol. Isolation of a single charge state at m/z 5,340 in the quadrupole and subsequent collision-induced dissociation (CID) in the gas-filled collision cell leads to the formation of ions from individual subunits and subcomplexes, identified by their mass and charge in the TOF analyzer. PMID- 11922311 TI - Molecular weight distributions of heavy aromatic petroleum fractions by Ag+ electrospray ionization mass spectrometry. AB - The ability of electrospray ionization mass spectrometry (ESI MS) to analyze heavy aromatic petroleum fractions using silver nitrate as a reagent compound to form characteristic adduct ions has been examined. The complexation of aromatic compounds containing long alkyl substituents with the silver ion leads to the formation of abundant adduct ions such as [M + Ag]+ and [2M + Ag]+. The concentration of the [2M + Ag]+ ions can be reduced by increasing the sampling cone voltage. Molecular ions and other adduct ions may also be formed depending on the structure of the aromatic molecule. Results obtained from the analysis of representative heavy petroleum fractions and vacuum residues by the Ag+ ESI MS method and conventional ionization methods were in good agreement. The current method extends the applicability of electrospray ionization to the analysis of neutral hydrocarbons in heavy aromatic petroleum fractions. It is simple and compatible with widely available LC/MS instrumentation. The extreme complexity of the Ag PMID- 11922312 TI - DNA fragment sizing by single molecule detection in submicrometer-sized closed fluidic channels. AB - The fabrication of fluidic channels with dimensions smaller than 1 microm is described and characterized in respect to their use for detection of individual DNA molecules. The sacrificial layer technique is used to fabricate these devices as it provides CMOS-compatible materials exhibiting low fluorescence background. It also allows creating microfluidics circuitry of submicrometer dimensions with great control. The small dimensions facilitate single molecule detection and minimize events of simultaneous passage of more than one molecule through the measurement volume. The behavior of DNA molecules inside these channels under an applied electrical field was first studied by fluorescence correlation spectroscopy using M13 double-stranded DNA. A linear relationship between the flow speed and applied electric field across the channel was observed. Speeds as high as 5 mm/s were reached, corresponding to only a few milliseconds of analysis time per molecule. The channels were then used to characterize a mixture of nine DNA fragments. Both the distribution and relative proportions of the individual fragments, as well as the overall concentration of the DNA sample, can be deduced from a single experiment. The amount of sample required for the analysis was approximately 10,000 molecules, or 76 fg. Other potential applications of these submicrometer structures for DNA analysis are discussed. PMID- 11922313 TI - Design and synthesis of Mg2+-selective fluoroionophores based on a coumarin derivative and application for Mg2+ measurement in a living cell. AB - Novel Mg2+ fluorescent molecular probes (KMG-20-AM and KMG-27-AM; where AM is an acetoxymethyl group) based on a coumarin possessing a charged beta-diketone structure were designed and synthesized. These fluorescent probes produced a red shift from 425 to 445 nm in the absorption spectra after formation of a complex with Mg2+. The fluorescence spectra of these probes also showed a red shift from 485 to 495 nm and an increasing fluorescence intensity after formation of a complex with Mg2+. The optimum experimental conditions were excitation wavelength of 445 nm and a monitored wavelength of 500 nm, where these probes functioned as an indicator showing an image of increasing fluorescence in the presence of Mg2+. These probes showed a "seesaw-type" fluorescent spectral change with the isosbestic point at 480 nm due to the light excitation at 445 nm, which indicates that ratiometry can be used for the measurement. The molecular probes formed a 1:1 complex with Mg2+ and the dissociation constant (Kd) was 10.0 mM for KMG-20. The association constants of the probes with Mg2- were approximately 3 times higher than that with Ca2+, which showed that the selectivity of Mg2+ versus Ca2+ for these probes was over 200 times higher than that for commercially available Mg2+ fluorescent molecular probes such as mag-fura-2, Magnesium Green. As an application of these probes, intracellular fluorescent imaging of Mg2+ was demonstrated using a fluorescent microscope. After the addition of KMG-20-AM and KMG-27-AM into PC12 cells, a strong fluorescence was observed in the cytoplasm and a weak fluorescence in the nuclei region. After treatment with a high-K+ medium, the fluorescence intensity increased due to increasing intracellular Mg2+. The real image of Mg2+ release from the magnesium store was successfully observed with these Mg2+ fluorescent probes. PMID- 11922314 TI - Effective time averaging of multiplexed measurements: a critical analysis. AB - Multiplexing and time averaging of signal are effective noise reduction protocols applied in many analytical measurement systems. The efficacy of these protocols may be reduced by random occurrences of high-magnitude noise that do not conform to the statistical distribution of noise for all other measurements in the data set. This high-magnitude noise, which may have an insignificant probability of occurrence for a single measurement, almost certainly affects data collected in a multichannel, multiplexed modality, such as Fourier transform infrared (FT-IR) spectroscopic imaging employing focal plane array detectors. To recover time averaging advantages in these cases, we present a general coaddition method that uses two statistical measures, the mean and median of the ensemble of measurements of a signal, to obtain a better estimate of the true signal than that estimated by time averaging alone. This method, termed median filtered time averaging, is shown to be an effective noise removal procedure for FT-IR imaging data. The effects of noise removal on time averaging and multiplexing are examined theoretically and are demonstrated for hyperspectral infrared microspectroscopic imaging data obtained from human skin biopsies by using a rapid data acquisition procedure. PMID- 11922315 TI - Immobilization of DNA hydrogel plugs in microfluidic channels. AB - Acrylamide-modified DNA probes are immobilized in polycarbonate microfluidic channels via photopolymerization in a polyacrylamide matrix. The resulting polymeric, hydrogel plugs are porous under electrophoretic conditions and hybridize with fluorescently tagged complementary DNA. The double-stranded DNA can be chemically denatured, and the chip may be reused with a new analytical sample. Conditions for photopolymerization, hybridization, and denaturation are discussed. We also demonstrate the photopolymerization of plugs containing different DNA probe sequences in one microfluidic channel, thereby enabling the selective detection of multiple DNA targets in one electrophoretic pathway. PMID- 11922316 TI - Biosensor for dengue virus detection: sensitive, rapid, and serotype specific. AB - A serotype-specific RNA biosensor was developed for the rapid detection of Dengue virus (serotypes 1-4) in blood samples. After RNA amplification, the biosensor allows the rapid detection of Dengue virus RNA in only 15 min. In addition, the biosensor is portable, inexpensive, and very easy to use, making it an ideal detection system for point-of-care and field applications. The biosensor is coupled to the isothermal nucleic acid sequence-based amplification (NASBA) technique with which small amounts of virus RNA are amplified using a simple water bath. During the NASBA reaction, a generic sequence is attached to all RNA molecules as described earlier (Wu, S. J.; Lee, E. M.; Putvatana, R.; Shurtliff, R. N.; Porter, K R.; Suharyono, W.; Watt, D. M.; King, C. C.; Murphy, G. S.; Hayes, C. G.; Romano, J. W. J. Clin. Microbiol. 2001, 39, 2794-2798.). It has been shown earlier that Dengue virus can be detected specifically using two DNA probes: a first probe hybridized with the attached generic sequence and, therefore, bound to every amplified RNA molecule; and a second probe either bound to all four Dengue virus serotypes or chosen to be specific for only one serotype. These probes were utilized in the biosensor described in this publication. For a generic Dengue virus biosensor, the second probe is complementary to a conserved region found in all Dengue serotypes. For identification of the individual Dengue virus serotypes, four serotype-specific probes were developed (Wu, S. J.; Lee, E. M.; Putvatana, R.; Shurtiff, R. N.; Porter, K. R.; Suharyono, W.; Watt, D. M.; King, C. C.; Murphy, G. S.; Hayes, C. G.; Romano, J. W. J. Clin. Microbiol. 2001, 39, 2794-2798.). The biosensor is a membrane-based DNA/RNA hybridization system using liposome amplification. The generic DNA probe (reporter probe) is coupled to the outside of dye-encapsulating liposomes. The conserved or Dengue serotype specific probes (capture probes) are immobilized on a polyethersulfone membrane strip. Liposomes are mixed with amplified target sequence and are then applied to the membrane. The mixture is allowed to migrate along the test strip, and the liposome-target sequence complexes are immobilized in the capture zone via hybridization of the capture probe with target sequence. The amount of liposomes present in the immobilized complex is directly proportional to the amount of target sequence present in the sample and can be quantified using a portable reflectometer. The different biosensor components have been optimized with respect to sensitivity and, foremost, specificity toward the different serotypes. An excellent correlation to a laboratory-based detection system was demonstrated. Finally, the assay was tested using a limited number of clinical human serum samples. Although Dengue serotypes 1, 2 and 4 were identified correctly, serotype 3 displayed low cross reactivity with biosensors designed for detection of serotypes 1 and 4. PMID- 11922317 TI - A programmable fragmentation analysis of proteins by in-source decay in MALDI-TOF mass spectrometry. AB - Here we describe an algorithm for identifying peptides/ proteins of known sequence and unknown peptides from partial spectra generated by an in-source decay (ISD) technique coupled with matrix-assisted laser desorption/ ionization time-of-flight mass spectrometry. The identification of protein fragments is processed with a software program called CMATCH, which generates candidate subsequences for both known peptides/proteins and unknown peptides for the major product ions in the spectral range m/z 400-5000 and then matches these to known protein sequences contained in a reference database for the known peptides/proteins. CMATCH, which is compiled for MSDOS or WINDOWS95/NT, has two main advantages: first, the candidate subsequences are generated automatically without the need for supplementary information concerning the distribution of either N-terminal or C-terminal ions in the spectra for both known peptides/proteins and unknown peptides; second, the highest coordinated homologous sequences are picked up automatically from the reference database as the best matches with known peptides/proteins. Examples from the ISD spectra of several test proteins demonstrate the efficacy of this protein identification software. PMID- 11922318 TI - UV resonance raman spectroscopic detection of nitrate and nitrite in wastewater treatment processes. AB - The 204- and 229-nm excited UV resonance Raman spectra of wastewater solutions containing sodium nitrite and nitrate were measured in the concentration range 7 microM to 3.5 mM (0.1-50 ppm nitrogen). The other chemical species present in wastewater do not interfere with Raman measurements of NO2-/NO3- bands. We observe detection limits of < 14 microM (< 200 ppb) for both NO2- and NO3-. UV resonance Raman spectroscopy appears to be an excellent tool for on-line monitoring of NO2-/NO3- in wastewater for the real-time control of water treatment plants. PMID- 11922319 TI - Pin-printed chemical sensor arrays for simultaneous multianalyte quantification. AB - A new approach to rapidly produce micrometer-scale sensor elements into reusable multianalyte chemical sensor arrays is demonstrated. By using pin printing technology in concert with sol-gel processing methods, we form discrete xerogel based microsensors on a planar substrate. We illustrate the new approach by forming discrete 02- and pH-responsive sensing elements into arrays that allow one to simultaneously determine O2 and pH in aqueous samples. The pin printing method allows one to prepare sensor elements that are on the order of 100 microm in diameter, 1-2 microm thick, at a rate of approximately one sensor element per second with a single pin. Within a given calibrated array, the sensor element-to sensor element response is reproducible to within 5%, the sensor element short- and long-term reproducibilities are 3 and 6%, respectively, and the array-to array response reproducibility is 11%. These results demonstrate the potential of this methodology for rapidly forming ensembles of reusable sensor arrays for simultaneous multianalyte detection. PMID- 11922321 TI - Virulence of recent isolates of Mycoplasma synoviae in turkeys. AB - Systemic Mycoplasma synoviae (MS) infection was induced experimentally in commercial turkeys with recent MS isolates (K4822D and K4774J) from turkey breeder flocks that exhibited no clinical signs typical of MS infection except for a low incidence of swollen footpads. The virulence of each strain was compared by evaluating gross and microscopic lesions, serologic responses, and MS isolation rates at 10 and 21 days postchallenge and by comparing these results with those obtained from a known virulent isolate (K1968), another previously characterized field isolate (K4463B), and unchallenged controls. All strains induced lesions typical of infectious synovitis but showed distinct differences in the extent of the gross and microscopic lesions and in the isolation rates from the tissues in turkeys. K1968 induced the most extensive lesions in hock and stifle joints and footpads, but strains K4822D, K4774J, and K4463B all induced synovitis and were similar in virulence for synovial tissues. Very mild respiratory lesions were induced by all of the strains studied. All strains yielded strong positive serologic responses. We concluded that these recent field isolates, although able to induce synovitis, are less virulent for turkeys than a known virulent strain. Nevertheless, under severe experimental challenge, these strains have the capability of causing lesions that may be incompatible with economical turkey production. PMID- 11922320 TI - Lymphocyte proliferation response during Eimeria tenella infection assessed by a new, reliable, nonradioactive colorimetric assay. AB - The application of a tetrazolium salt, WST-8,2-(2-methoxy-4-nitrophenyl)-3-(4 nitrophenyl)-5-(2,4-disulfophenyl)-2H tetrazolium, monosodium salt to the lymphocyte proliferation assay in the chicken system was evaluated. Proliferation of concanavalin (Con A)-induced splenic lymphocytes and peripheral blood lymphocytes (PBL) was evaluated with WST-8 and 3-(4,5-dimethylthiazol-2-yl)-2,5 diphenyltetrazolium bromide (MTT). Coefficients of correlation (r) between these two reagents were 0.98 and 0.97 in splenic lymphocytes and PBL, respectively. In general, the sensitivity of the WST-8 assay was significantly higher than that of the MTT assay, and the standard deviations of the WST-8 assay were significantly lower than those of the MTT assay. The WST-8 assay was fast and highly reproducible and provided a good indication of mitogen-induced proliferation of spleen cells induced by Con A. With the use of the WST-8 assay, splenic mitogenic response of chickens infected with Eimeria decreased transiently at 7 days but increased significantly at 10 days after primary infection compared with that of uninfected chickens. Additionally, the measurement of interleukin (IL)-2 production with WST-8 was highly reproducible and showed a significant increase in IL-2 production upon stimulation of Eimeria tenella-immune spleen cells with Con A. After E. tenella infection, splenic IL-2 production increased significantly at 7 days post-primary and at 2 days post-secondary infection. The WST-8 assay is fast, simple, and more reproducible and sensitive than the MTT assay. This study demonstrates the effectiveness of the WST-8 assay to assess cell-mediated immune response of chickens in normal and disease states. PMID- 11922322 TI - Isolation from turkey breeder hens of a reassortant H1N2 influenza virus with swine, human, and avian lineage genes. AB - Type A influenza viruses can infect a wide range of birds and mammals, but influenza in a particular species is usually considered to be species specific. However, infection of turkeys with swine H1N1 viruses has been documented on several occasions. This report documents the isolation of an H1N2 influenza virus from a turkey breeder flock with a sudden drop in egg production. Sequence analysis of the virus showed that it was a complex reassortant virus with a mix of swine-, human-, and avian-origin influenza genes. A swine influenza virus with a similar gene complement was recently reported from pigs in Indiana. Isolation and identification of the virus required the use of nonconventional diagnostic procedures. The virus was isolated in embryonated chicken eggs by the yolk sac route of inoculation rather than by the typical chorioallantoic sac route. Interpretation of hemagglutination-inhibition test results required the use of turkey rather than chicken red blood cells, and identification of the neuraminidase subtype required the use of alternative reference sera in the neuraminidase-inhibition test. This report provides additional evidence that influenza viruses can cross species and cause a disease outbreak, and diagnosticians must be aware that the variability of influenza viruses can complicate the isolation and characterization of new isolates. PMID- 11922323 TI - Changes in serum ovotransferrin levels in chickens with experimentally induced inflammation and diseases. AB - A competitive enzyme immunoassay was developed to measure the changes in serum levels of ovotransferrin (OTF) during inflammation and infectious diseases in chickens. The assay is based on the competition of serum OTF with a fixed concentration of biotin-labeled OTF to bind to a rabbit anti-chicken transferrin antibody immobilized on microtiter wells. After several washing steps, the antibody-bound biotinylated OTF is probed with streptavidin-horseradish peroxidase conjugate (HRP) followed by a colorimetric detection of the HRP activity. The relative changes in the optical density of color are plotted against the competing concentrations of OTF with logarithmic regression to generate a standard curve that is used to determine the concentrations of OTF in unknown samples. Serum had no effect on the measurement of OTE By this method, the time course changes of serum OTF levels in 4-wk-old male broiler chickens that were subjected to inflammation by croton oil injection were measured. The results showed croton oil-induced inflammation elevated serum OTF levels at 16 hr postinjection. OTF levels reached a peak by 72 hr, remained high through 120 hr, and returned to a basal level of olive oil-injected controls by 240 hr. There were no changes in serum OTF levels at any of the above time points in olive oil injected control chickens. For studies with poultry diseases, specific-pathogen free (SPF) male chickens were challenged with known bacterial and viral pathogens, and serum was collected at the height of the infection, i.e., 7 days after the challenge. Compared with uninjected controls, the SPF chickens challenged with Escherichia coli, fowl poxvirus, respiratory enteric orphan virus, infectious bursal disease virus, infectious bronchitis virus, or infectious laryngotracheitis virus had higher levels of OTF in serum. Inflammation-induced changes in serum OTF levels were also evident in the changes in the density of a 65-kD band protein corresponding to OTF. These results demonstrate that serum OTF may be a nonspecific clinical marker of inflammation associated with traumatic or infectious avian diseases. PMID- 11922324 TI - Distribution of staphylococcal enterotoxin genes among Staphylococcus aureus isolates from poultry and humans with invasive staphylococcal disease. AB - Food poisoning by Staphylococcus aureus affects hundreds of thousands of people each year. Staphylococcus aureus also causes invasive diseases such as arthritis (in poultry) and septicemia (in poultry and humans). Foodborne disease is caused by the ingestion of a staphylococcal enterotoxin (SE). Enterotoxin has also been associated with other S. aureus illnesses in humans and domestic animals. In this study, polymerase chain reaction was used to detect the staphylococcal enterotoxin genes, SEA, SEB, SEC, SED, and SEE, in S. aureus isolates associated with invasive disease in poultry and humans. In the 34 poultry isolates, only one isolate was found to contain a SE gene, sec. In the 41 human isolates, over 51% tested positive for an SE gene with 12.2% positive for the gene for SEA, 2.4% for SEB, 22% for SEC, 24.4% for SED, and 0 for SEE. The disparity between the rates for SE gene(s) in poultry and human isolates suggests a lesser role for the enterotoxins in invasive poultry disease than in human disease. PMID- 11922325 TI - Serologic detection of experimental Salmonella enteritidis infections in laying hens by fluorescence polarization and enzyme immunoassay. AB - Detection of infected poultry flocks is essential for controlling eggborne transmission of Salmonella enteritidis to humans. The present study evaluated the detection of antibodies in the sera of experimentally infected chickens by a fluorescence polarization assay with a tracer prepared from the O-polysaccharide of S. enteritidis and an enzyme-linked immunosorbent assay (ELISA) with an S. enteritidis flagellin antigen. In two trials, groups of specific-pathogen-free laying hens were infected orally with either 10(6) or 10(8) colony-forming units (CFU) of S. enteritidis (phage type 13a) or with 10(8) CFU of Salmonella typhimurium. Serum samples were collected before inoculation and at five subsequent weekly intervals. Both assays successfully detected the majority of hens infected with S. enteritidis at either dose level, but they also identified a substantial number of hens infected with S. typhimurium as seropositive. The fluorescence polarization test detected S. enteritidis infection significantly more often and cross-reacted with sera from hens infected with S. typhimurium significantly less often than the ELISA. The fluorescence polarization assay also offered advantages in terms of speed and methodologic simplicity. PMID- 11922326 TI - Pathotypes of avian Escherichia coli as related to tsh-, pap-, pil-, and iuc-DNA sequences, and antibiotic sensitivity of isolates from internal tissues and the cloacae of broilers. AB - One hundred four Escherichia coli isolates were collected from internal tissues and the cloacae of broilers with colibacillosis or from the cloacae of healthy birds. The isolates were tested for the presence of DNA sequences for temperature sensitive hemagglutinin (tsh), for P (pap) and F1 (pil) fimbriae, and for aerobactin synthesis (iuc) by DNA/DNA hybridization. The isolates were also tested for O1, O2, and O78 serogroups, serum and antibiotic resistance, and virulence in day-old chickens. The Tsh/Pil/Iuc was the major pathotype detected in 53.8% of isolates from internal tissues, as compared with only 28.8% of isolates from the cloacae. The Tsh/Pap/Iuc pathotype was detected at a lower frequency (15.4%) but only in isolates from internal tissues. Among the virulence associated marker genes, tsh and iuc were detected in most of the isolates from internal tissues (90.4% and 92.3%), as compared with only 51.9% and 63.5% of isolates from the cloacae, respectively, pap was detected to a lesser extent, in 25% of isolates but only from internal tissues. In contrast to the pil gene, the tsh-, pap-, and iuc-DNA sequences were more frequently detected in isolates from internal tissues than in isolates from the cloacae. O-antigen typing revealed that 25% of isolates belonged to serogroups O1 (4.8%), O2 (9.6%), and O78 (10.6%). Although most isolates appeared to be resistant to serum, only isolates from internal tissues were virulent in day-old chickens in contrast to isolates from the cloacae. More than 10% of isolates were resistant to most of the antibiotics used for the study. However, less resistance to enrofloxacin and norfloxacin was observed. Our data suggest that the Tsh/Pil/Iuc and Tsh/Pap/Iuc pathotypes and Tsh and Iuc virulence-associated markers are important factors of avian pathogenic E. coli. Enrofloxacin appeared to be the best choice for treatment of the infection. PMID- 11922327 TI - Pathogenic and immunosuppressive effects of avian pneumovirus in turkeys. AB - Avian pneumovirus (APV) causes a respiratory disease in turkeys. The virus has been associated with morbidity and mortality due to secondary infections. Our objective was to determine if APV caused immunosuppression in the T-cell or B cell compartments and to study the pathogenesis of the disease in APV maternal antibody-lacking 2-wk-old commercial turkeys. APV was administered by the eyedrop/intranasal route. Observations were made for gross lesions, viral genome, and T-cell mitogenesis and cytokine secretion at 3, 5, 7, 14, and 21 days postinoculation (DPI). During the acute phase of the disease that lasted for about 1 wk, the turkeys exposed to APV showed clinical signs characterized by nasal discharge and sinus swelling. Virus genome was detected by in situ hybridization in cells of turbinates and trachea at 3 and 5 DPI. At 3 and 5 DPI, spleen cells of the birds infected with APV markedly decreased proliferative response to concanavalin A (Con A). Con A and lipopolysaccharide stimulation of spleen cells from virus-exposed turkeys resulted in accumulation of nitric oxide inducing factors (NOIF) in the culture fluid. NOIF were not detected in culture fluids of Con A-stimulated spleen cells of virus-free turkeys. APV did not compromise the antibody-producing ability of turkeys against several extraneous antigens such as Brucella abortus and tetanus toxoid. PMID- 11922328 TI - Development, characterization, and preliminary evaluation of a temperature sensitive mutant of Ornithobacterium rhinotracheale for potential use as a live vaccine in turkeys. AB - A temperature-sensitive (Ts) mutant strain of Ornithobacterium rhinotracheale (ORT) was developed after exposure of the wild-type organism to N-methyl-N'-nitro N-nitrosoguanidine. The Ts mutant strain grew at 31 C but had its growth inhibited at 41 C unlike wild-type parent strain. The Ts mutant and parent strains were characterized. Morphologic and biochemical properties of wild-type and mutant strains did not show any differences. The strains were also characterized by polymerase chain reaction (PCR)-based fingerprinting methods. Results showed similar patterns in repetitive sequences by repetitive PCR (enterobacterial repetitive intergenic consensus, highly conserved repeated DNA elements present in Streptococcus pneumoniae (BOX), repetitive extragenic palindromic, and Salmonella enteritidis repetitive element primers); however, random amplified polymorphic DNA fingerprinting was able to differentiate mutant and parent strains showing a unique pattern for each of the ORT strains. The rationale for the use of a Ts strain as a vaccine is based on the ability of the mutant to colonize the upper respiratory tract but not the lower respiratory tract and systemic system of the birds, where the wild-type strain causes severe lesions. In a preliminary evaluation, Ts strain of ORT was recovered from tracheas and choanae of Ts-treated turkeys for 13 days postadministration of the strain either in drinking water or by oculonasal instillation. Humoral immune response was detected in Ts-vaccinated but not in control group birds after 3 wk postadministration. Results suggest that Ts strain of ORT has promising potential use as a live vaccine for ORT. PMID- 11922329 TI - Pathogenesis and tissue distribution of a variant strain of infectious bursal disease virus in commercial broiler chickens. AB - The detection of either infectious virus, viral antigen, and/or viral RNA in different tissues of commercial broilers inoculated at 1 day of age with E/Del variant strain of infectious bursal disease virus (IBDV) was investigated at 2, 4, and 6 wk postinoculation (PI). Virus was readily isolated from homogenates of bursa, cecal tonsils, and bone marrow at 2 and 4 wk PI. Virus isolation coupled with immunoperoxidase assay or reverse transcription-polymerase chain reaction for IBDV-specific RNA extended the window of IBDV detection in the bursa of Fabricius to 6 wk PI. Serology indicated an active early virus infection; however, viral pathology was observed later and beginning at 4 wk PI. This study indicates that variant strains of IBDV may be present in commercial broilers longer than previously thought, and cecal tonsils and bone marrow may serve as nonbursal lymphoid tissues supporting virus replication at later time points PI. PMID- 11922330 TI - Results of salmonella isolation from poultry products, poultry, poultry environment, and other characteristics. AB - Five hundred sixty-nine Salmonella were isolated out of 4745 samples from poultry products, poultry, and poultry environment in 1999 and 2000 from the Pacific northwest. These Salmonella were identified to their exact source, and some were serogrouped, serotyped, phage typed, and tested for antibiotic sensitivity. Food product samples tested included rinse water of spent hens and broilers and chicken ground meat. Poultry environment samples were hatchery fluff from the hatcheries where eggs of grandparent broiler breeders or parent broiler breeder eggs were hatched and drag swabs from poultry houses. Diagnostic samples were of liver or yolk sac contents collected at necropsy from the young chicks received in the laboratory. Of these samples tested, 569 were Salmonella positive (11.99%). Ninety-two Salmonella were serogrouped with polyvalent somatic antisera A-I and the polymerase chain reaction. Somatic serogroups B and C comprised 95.25% of all the Salmonella. Out of a total of 569 positive samples, 97 isolates of Salmonella were serotyped. A total of 16 serotypes and an unnamed Salmonella belonging to serogroup C1 were identified. The Salmonella serotypes were heidelberg (25.77%); kentucky (21.64%); montevideo (11.34%); hadar and enteritidis (5.15% each); infantis, typhimurium, ohio, and thompson (4.12% each); mbandaka and cerro (3.09% each); senftenberg (2.06%); berta, istanbul, indiana, and saintpaul (1.03% each); and an unnamed monomorphic Salmonella (2.06%). Ninety two Salmonella were tested for drug sensitivity with nine different antimicrobials. All of the 92 Salmonella were resistant to erythromycin, lincomycin, and penicillin except one sample (S. berta), which was moderately sensitive to penicillin. All of the tested Salmonella were susceptible to sarafloxacin and ceftiofur. The percentages of Salmonella susceptible to sulfamethoxazole-trimethoprim, gentamicin, triple sulfa, and tetracycline were 97.83%, 92.39%, 86.96%, and 82.61%, respectively. PMID- 11922331 TI - Minimization of pathologic changes in Ornithobacterium rhinotracheale infection in turkeys by temperature-sensitive mutant strain. AB - The protection elicited by a temperature-sensitive (Ts) mutant of Ornithobacterium rhinotracheale (ORT) vaccine against challenge with pathogenic strain was investigated. In Experiment 1, specific serologic response to ORT was detected in 12%-19% of Ts-vaccinated birds at 3 wk postvaccination by either drinking water or oculo-nasal instillation. At 7 days postchallenge, 100% of Ts vaccinated turkeys of all groups were able to respond with an ORT-specific antibody response, but the control group was not, suggesting the potential of Ts strain to evoke immune protection. The study also revealed a statistically significant ability of the Ts strain to protect vaccinated turkeys against gross lesions caused by the pathogenic strain of ORT in treated groups vs. control. In Experiment 2, seroconversion was detected by enzyme-linked immunosorbent assay in birds after they were given the Ts strain in drinking water in field conditions. The results of the field study showed mean scores of gross lesions of nonvaccinated/challenged groups to be up to seven times higher than those of the vaccinated/challenged group. In addition, reisolation rates and quantification of ORT colonies per gram of lung tissue were significantly lower for vaccinated/challenged than for nonvaccinated/challenged turkeys. In conclusion, results from laboratory and field experiments suggest that use of the Ts mutant strain of ORT as a live vaccine would be a suitable method to evoke protection against ORT infection in turkeys. PMID- 11922332 TI - Virulence analysis of a Salmonella gallinarum strain by oral inoculation of 20 day-old chickens. AB - In order to know the effect of in vitro passages on the pathogenicity of the Salmonella gallinarum strain INTA 91, a lyophilized culture was compared with the same strain recently isolated from a sick bird. The mean lethal dose (LD50) of the orally administered lyophilized culture was determined as 2.04 x 10(8) colony forming units (CFU)/chicken. There was no correlation between the LD50 dose and the degree of disease produced; doses 10 or 100 times higher than the calculated LD50 did not produce a more severe disease. In trial 1, chickens were challenged with 1.02 x 10(9) CFU per chicken (5LD50) of the lyophilized strain and reached 52.2% mortality at the end of the assay. In trial 2, three different groups of chickens were infected with a recent isolate of the same strain: 2.04 x 10(8) CFU/chicken, 4.1 x 10(8) CFU/chicken, and 2.1 x 10(9) CFU/chicken (i.e., 1LD50, 2LD50, and 10LD50 of the dose calculated for the lyophilized strain, respectively). These chicken groups presented higher mortality rates (90%, 100%, and 95%, respectively) than previous trials, showing that the S. gallinarum strain used here increased its virulence by in vivo infected chicken passage. In all assays, the disease started after an incubation period of around 5-6 days. To obtain reliable and reproducible results in future challenge experiments, a fixed limited number of in vitro passages of the S. gallinarum strain must be determined. PMID- 11922334 TI - Effect of a commercial competitive exclusion culture on reduction of colonization of an antibiotic-resistant pathogenic Escherichia coli in day-old broiler chickens. AB - One-day-of-age broiler chickens were administered a commercial competitive exclusion (CE) product and then challenged by three different methods with an Escherichia coli O78:K80 that was pathogenic for poultry and resistant to six antibiotics. Three challenge methods were used on 2-day-old broilers: direct challenge, precolonized seeder, and instant seeder. Direct challenge was accomplished by administering the challenge E. coli per os. The precolonized seeder challenge had two chicks that had received the challenge E. coli 24 hr previously, whereas the instant seeder challenge had two chicks given the challenge E. coli per os with immediate placement with the experimental birds. One oral dose of the commercial CE product significantly reduced the colonization of the small intestine, large intestine, and ceca by the highly antimicrobial resistant poultry pathogenic E. coli O78:K80 at 7 and 14 days postchallenge by all three challenge methods. The overall mean reductions in colonization were 3.0 log10 for the large intestine, 3.0 log10 for the small intestine, and 4.0 log10 for the cecum. The most severe challenge method, on the basis of the least amount of reduction of colonization of the challenge E. coli by the CE, was by the direct oral gavage at 2 days of age. PMID- 11922333 TI - Histopathologic and bacteriologic evaluations of cellulitis detected in legs and caudal abdominal regions of turkeys. AB - The objective of this study was to identify the causative agent of cellulitis in turkeys. Eighteen flocks from nine producers were sampled at the local processing plant, and 37 birds with cellulitis on legs or caudal thoracic area were obtained. None of the 37 birds with cellulitis had lesions in other organs. On gross examination, lesions were categorized into two groups: cellulitis with unopened skin lesions (type a) and cellulitis with opened skin lesions (type b). Histopathologically, cellulitis with unopened skin lesions had dermal necrosis with underlying fibrin and inflammatory exudate but cellulitis with open skin lesions had chronic granulomatous/granulation tissue-type reaction associated with foreign material. A complete bacteriologic study was conducted on 25 of 37 birds. Bacteria were isolated from 12 of the 25 birds with cellulitis lesions. No aerobic, microaerophilic, or anaerobic bacteria were isolated from the remaining 13 birds with cellulitis lesions. Escherichia coli was isolated in low numbers in mixed cultures with Proteus mirabilis, Lactobacillus spp., Klebsiella spp., and Staphylococcus spp. in 9 of 12 lesions. The remaining few cases yielded P. mirabilis in pure culture or in mixed culture with Pseudomonas aeruginosa. Types a and b cellulitis lesions in turkeys could be associated with primary contact dermatitis and skin abrasions, respectively. Their occurrence is likely associated with different management practices. PMID- 11922335 TI - Interleukin-2 production in SC and TK chickens infected with Eimeria tenella. AB - SC and TK inbred chicken strains display differential protective immunity to coccidiosis, SC being more resistant and TK susceptible to disease. In this study, the association between interleukin (IL)-2 and disease phenotype was assessed by cytokine quantification in serum, duodenum, cecum, and spleen cell cultures of SC and TK chickens experimentally infected with Eimeria tenella. In general, after primary infection, SC and TK strains produced equivalent amounts of IL-2 in all sources examined. However, after secondary infection, SC animals displayed significantly greater IL-2 levels in serum and the duodenum compared with strain TK. IL-2 production after reinfection with Eimeria may be an important factor contributing to the genetic differences in coccidiosis between SC and TK chickens and provides a rational foundation for cytokine-based immunotherapeutic approaches to disease control strategies. PMID- 11922336 TI - Performance and oocyst shedding in broiler chickens orally infected with Cryptosporidium baileyi and Cryptosporidium meleagridis. AB - To study effects of experimental cryptosporidiosis, broiler chickens were infected per os with 5 x 10(5) oocysts of Cryptosporidium baileyi and Cryptosporidium meleagridis. In the first experiment, chickens were infected with oocysts of C. baileyi at the age of 7, 14, and 21 days. In the second experiment, chickens were infected with oocysts of C. baileyi, C. meleagridis, or both cryptosporidial species at the age of 7 days. Although clinical signs of infection were apparent, neither final live weight nor mortality was significanty influenced in chickens infected with a single Cryptosporidium species. In chickens infected with C. meleagridis, the growth retardation was observed in the 2-wk period after infection. The compensatory growth, however, started when the oocyst shedding had ceased. The number of oocysts in excreta specimens of chickens infected with C. meleagridis was two to three times lower than in excreta of chickens infected with C. baileyi. Chickens infected with both C. baileyi and C. meleagridis (5 x 10(5) oocysts of each) had significantly lower final live weight and worse feed efficiency than chickens of other groups. Concurrent infection did not influence individual C. baileyi or C. meleagridis oocyst shedding. PMID- 11922337 TI - Field evaluation of tylosin premix in layers previously vaccinated with a live Mycoplasma gallisepticum vaccine. AB - Mycoplasma gallisepticum infection results in numerous clinical signs including a reduction in egg production in laying chickens. Attempts to prevent mycoplasmosis have included vaccination with both killed and attenuated live M. gallisepticum strains. Live vaccines provide reduction in clinical signs and have been shown to replace indigenous strains when used in a consistent program for several placements. Antibiotic therapy is another option for controlling losses associated with mycoplasmosis. Therapeutic antibiotics with activity against mycoplasma approved for use in poultry include tetracyclines and tylosin. These drugs also are approved for feed efficiency when administered in the feed at levels below the therapeutic index for mycoplasma. The data presented here suggest that birds vaccinated with the live 6/85 strain of M. gallisepticum and then fed tylosin, at the approved level for feed efficiency, exhibit a serologic vaccine response similar to that of unmedicated birds but show improved feed efficiency. PMID- 11922338 TI - In vitro antibiotic susceptibility of field isolates of Mycoplasma synoviae in Argentina. AB - Minimum inhibitory concentrations (MICs) were determined in vitro for 7 antibiotics (aivlosin, enrofloxacine, tylosin, tiamulin, kitasamycin, chlortetracycline, and oxytetracycline) against eight recent local Argentinean isolates and two standard strains of Mycoplasma synoviae. Aivlosin (3-acetyl-4" isovaleryl tylosin tartrate), tylosin, and tiamulin showed the lowest MICs with MIC90s of 0.006, 0.012, and 0.05 microg/ml, respectively. Except one strain that showed resistant values to chlortetracycline (> or = 12.5 microg/ml), all the analyzed strains were susceptible in different degrees to all the antibiotics tested. In this study, the improved activity of the tylosin-derived drug, aivlosin, was confirmed because it showed, in most strains, MIC values half those for tylosin. PMID- 11922339 TI - Direct polymerase chain reaction detection of Campylobacter spp. in poultry hatchery samples. AB - A rapid, sensitive, and specific polymerase chain reaction (PCR) assay was developed for the direct detection of Campylobacter in environmental samples from hatcheries. PCR, with a set of primers specific for the Campylobacter flaA short variable region (SVR), detected the presence of Campylobacter in both fluff and eggshell samples; however, a determination of whether the organism was living or dead could not be made. Conventional cultural methods detected no Campylobacter from the same samples. An additional benefit of the direct PCR assay is it allows for the production of a product that can be sequenced to provide further epidemiologic information. PMID- 11922340 TI - The influence of stress from transport and handling on hematologic and clinical chemistry blood parameters of racing pigeons (Columba livia domestica). AB - In the present study, the influence of stress from handling and transport on some frequently examined blood parameters of racing pigeons was evaluated. After 3 hr, there was a highly significant (P < 0.01) increase in the number as well as in the percentage of heterophils and decrease of lymphocytes. In clinical chemistries, increases of creatine kinase and glucose and a decrease of uric acid were observed. There was a mean decrease of the total white blood count of >15% that was less significant (P < 0.05). Changes in lactate dehydrogenase, basophils, and monocytes did not prove to be significant; eosinophils, aspartate aminotransferase, total protein, and the packed cell volume were not influenced by stress. PMID- 11922342 TI - Four-legged broiler chicken with two cloacae and three ceca. AB - This case report examines an Arbor Acres female x Peterson male crossbred 6-wk old female broiler chicken with four legs, two cloacae, and three ceca. The intestines occupied the caudodorsal portion of the abdominal cavity with three ceca attached to the terminal end of the ileum. The left lateral cecum was larger and had a divided distal end that terminated into 2-cm-long blind sacs. The rectum was dilated and divided into two cloacae that each opened on the dorsolateral aspect of both the right and left cloacal elevator muscles. The two extra legs were attached to the pygostyle by cartilage and skin tissue. Both femoral heads of the extraneous legs were fused together. The extra legs were smaller in size (approximately 17 cm in length) compared with the normal legs (approximately 22 cm). The femoral heads of the two normal legs were disarticulated from the acetabulum. PMID- 11922341 TI - Fowl adenovirus serotype 4 associated with outbreaks of infectious hydropericardium in Haryana, India. AB - During 1998, hydropericardium syndrome was observed among 3-to-6-wk-old broilers in 45 different flocks of Haryana, India, with mortality ranging between 10% and 30%. Fowl adenovirus (FAV) was isolated from one of the affected flocks by chicken embryo liver cell culture. Serum neutralization test and polymerase chain reaction assay coupled with restriction enzyme analysis confirmed that the isolated virus belonged to FAV serotype 4. The disease was reproduced in 28-day old broilers by subcutaneous and oral inoculation of isolated FAV4 alone. Typical hydropericardium and basophilic intranuclear inclusions in hepatocytes were observed in experimental birds by day 4 postinoculation. PMID- 11922343 TI - Neonatal avian pneumovirus infection in commercial turkeys. AB - Eleven market turkey flocks developed a respiratory disease characterized by coughing, swollen sinuses and nasal discharge. These symptoms first appeared between 3 and 16 days of age. Avian pneumovirus (APV) RNA was detected by reverse transcriptase (RT)-polymerase chain reaction (PCR) in six of six flocks tested. APV was detected by immunohistochemistry in turbinates of three of three affected flocks tested. Virus isolation attempts were negative. Ten of 11 flocks became seropositive on the APV enzyme-linked immunosorbent assay. Five weeks prior to hatch of these affected market turkeys, several breeder flocks in one geographic area had developed clinical signs and experienced decline in egg production typical of APV infection. In two breeder flocks, acute and convalescent sera indicated APV infection during the period of declining egg production. Attempts to detect APV RNA by RT-PCR from choanal cleft swabs of newly hatched poults were successful. Attempts to isolate the virus from these PCR-positive samples were negative. PMID- 11922344 TI - Avulsion of the common retinaculum in meat turkeys. AB - An increase in leg condemnations was experienced in several commercial meat flocks of male and female turkeys. Affected legs had hematomas around and above the tibiotarsus-tarsometatarsus (hock) joint. Avulsion of the common retinaculum from the external condyle was observed when the skin and muscular fascia were removed. A farm investigation was performed and similar changes were observed in turkeys between 10 and 19 wk of age. Interestingly, birds with avulsion of this fibrinous bridge were not lame. Body weights were normal, no infectious agent was detected, and mineral contents of bones and tendons were within normal limits. Avulsion of the retinaculum might not have been diagnosed in the past because the birds are not lame and the lesion is tightly covered by the skin and muscular fascia. PMID- 11922345 TI - Haemoproteus lophortyx infection in bobwhite quail. AB - This report chronicles recurring outbreaks of Haemoproteus lophortyx infection in captive bobwhite quail. Clinically, the signs of infection included reluctance to move, ruffled appearance, prostration, and death. These signs were associated with parasitemia, anemia, and the presence of large megaloschizonts in skeletal muscles, particularly those of the thighs and back. The average cumulative mortality for flocks experiencing outbreaks was over 20%. In a typical outbreak, mortality rose when the birds were 5-6 wk of age, peaked in 8-to-10-wk-old quail, and declined rapidly when the quail were 9-11 wk old. Outbreaks occurred exclusively between the months of May and October, and warm weather was determined to be a risk factor for H. lophortyx mortality. This protozoan most likely overwinters in native California quail in the area and is transmitted to quail on the ranch by an insect vector that emerges in warm weather. Infection of the large population of naive bobwhite quail on the ranch leads to amplification of H. lophortyx, resulting in epidemics in successive flocks. PMID- 11922346 TI - Salmonella enterica serovar enteritidis burden in broiler breeder chicks genetically associated with vaccine antibody response. AB - The relationship between antibody response to Salmonella enteritidis vaccine and internal organ burden of S. enteritidis is not fully understood. The genetic relationship, therefore, between postchallenge S. enteritidis burden and antibody response to S. enteritidis vaccine was determined in broiler breeder chicks. Sibling chicks from a broiler breeder male line were either inoculated with a pathogenic S. enteritidis or vaccinated with a commercial S. enteritidis vaccine. Spleen, liver, cecal wall, and cecal content samples from S. enteritidis challenged chicks (n = 120) were cultured for enumeration of bacteria. Unchallenged chicks (n = 314) were vaccinated at 11 days of age, and serum samples were taken at 10 days postvaccination. Antibody response to vaccination and number of S. enteritidis in cecal content cultures were negatively correlated (-0.772), demonstrating that genetic potential for greater antibody response to S. enteritidis vaccine is associated with lesser S. enteritidis bacterial burden in cecal content of broiler breeder chicks. The findings suggest that genetic selection for vaccine antibody responsiveness can lower bacterial burden in the gut lumenal content and, thus, potentially reduce contamination of poultry products at processing. PMID- 11922347 TI - Myxoid leiomyosarcoma of the gizzard in a broiler chicken. AB - A leiomyosarcoma was found in the gizzard of a 57-day-old female broiler chicken weighing 1.8 kg. Grossly, the tumor mass, 13.0 x 8.5 x 10.0 cm, enveloped the gizzard and had a gelatinous appearance due to the rich production of mucin. Miliary metastatic tumors were noted in the liver. Histopathologically, there was marked production of mucus throughout the tumor tissue, and densely or loosely arranged long spindle-shaped leiomyosarcoma cells proliferated. The tumor cells had a low rate of mitosis, showed slight cellular atypia, and, immunohistochemically, were positive for actin, alpha-smooth muscle actin, and desmin. Electron microscopically, various amounts of microfibrils with focal densities, dense patches, and basal plates were observed. PMID- 11922348 TI - Isolation of a reovirus from poult enteritis and mortality syndrome and its pathogenicity in turkey poults. AB - Poult enteritis and mortality syndrome (PEMS) is an acute, infectious intestinal disease of turkey poults, characterized by high mortality and 100% morbidity, that decimated the turkey industry in the mid-1990s. The etiology of PEMS is not completely understood. This report describes the testing of various filtrates of fecal material from control and PEMS-affected poults by oral inoculation into poults under experimental conditions, the subsequent isolation of a reovirus, ARV CU98, from one of the PEMS fecal filtrates, and in vivo and in vitro studies conducted to determine the pathogenicity of ARV-CU98 in turkey poults. In order to identify a filtrate fraction of fecal material containing a putative etiologic agent, poults were challenged in two independent experiments with 220- and 100-nm filtrates of fecal material from PEMS-negative and PEMS-positive poults. The 100 nm filtrate was chosen for further evaluation because poults inoculated with this filtrate exhibited mortality and significantly lower (P < or = 0.05) body weight and relative bursa weight, three clinical signs associated with PEMS. These results were confirmed in a third experiment with 100-nm fecal filtrates from a separate batch of PEMS fecal material. In Experiment 3, body weight and relative bursa and thymus weights were significantly lower (P < or = 0.05) in poults inoculated with 100-nm filtrate of PEMS fecal material as compared with poults inoculated with 100-nm filtrate of control fecal material. Subsequently, a virus was isolated from the 100-nm PEMS fecal filtrate and propagated in liver cells. This virus was identified as a reovirus on the basis of cross-reaction with antisera against avian reovirus (FDO strain) as well as by electrophoretic analysis and was designated ARV-CU98. When inoculated orally into poults reared under controlled environmental conditions in isolators, ARV-CU98 was associated with a higher incidence of thymic hemorrhaging and gaseous intestines. In addition, relative bursa and liver weights were significantly lower (P < or = 0.05) in virus-inoculated poults as compared with controls. Virus was successfully reisolated from virus-challenged poults but not from control birds. Furthermore, viral antigen was detected by immunofluorescence in liver sections from virus-challenged poults at 3 and 6 days postinfection and virus was isolated from liver at 6 days postinfection, suggesting that ARV-CU98 replicates in the liver. In addition to a decrease in liver weight, there was a functional degeneration as indicated by altered plasma alanine aminotransferase and aspartate aminotransferase activities in virus poults as compared with controls. Although this reovirus does not induce fulminating PEMS, our results demonstrated that ARV-CU98 does cause some of the clinical signs in PEMS, including intestinal alterations and significantly lower relative bursa and liver weights. ARV-CU98 may contribute directly to PEMS by affecting the intestine, bursa, and liver and may contribute indirectly by increasing susceptibility to opportunistic pathogens that facilitate development of clinical PEMS. PMID- 11922349 TI - Virulence factors of Escherichia cofi from cellulitis or colisepticemia lesions in chickens. AB - This study was designed to compare virulence factors of cellulitis-derived Escherichia coli to colisepticemic E. coli in order to clarify whether E. coli associated with cellulitis comprise a unique subset of pathogenic E. coli. Isolates were tested for serotype, capsule, aerobactin production, colicin production, the presence of the iss gene, and serum resistance. Untypable isolates made up the greatest percentage of each group. Serotypes O2 and O78 were the most commonly identified among both groups of isolates. No statistical differences in the distribution of aerobactin or colicin production, capsule, or iss gene were observed between groups. Cluster analysis showed that 90% of the E. coli isolates had greater than 42% livability in serum-resistance tests. No separation of colisepticemic vs. cellulitis E. coli isolates was observed on the basis of SR. Colicin production by E. coli was highly correlated with serum resistance (P = 0.0029). These data suggest that cellulitis E. coli have virulence traits similar to those of colisepticemic E. coli. PMID- 11922350 TI - Detection of infectious laryngotracheitis virus in formalin-fixed, paraffin embedded tissues by nested polymerase chain reaction. AB - Infectious laryngotracheitis virus (ILTV) is routinely diagnosed by histopathologic examination of trachea, eyelid, and lung tissues. Lesions consistent with infectious laryngotracheitis (ILT) infection include syncytial cell formation with intranuclear inclusion bodies. These changes are present during the acute phase of infection. To increase the sensitivity of detecting ILT, a nested polymerase chain reaction (PCR) was developed for detection of ILTV DNA. Nested PCR assay was specific for the amplification of ILTV DNA and did not amplify a variety of other avian pathogens. To further validate the ability of this assay to detect ILT, nested PCR was performed in formalin-fixed, paraffin embedded tissues from 35 cases of respiratory disease. Of the 35 cases, 12 were considered ILT suspects on the basis of initial clinical observation. Eleven of the 12 ILT-suspect cases were diagnosed as ILT, and the remaining 24 were diagnosed as nonspecific tracheitis (NST) by histopathologic examination. Histopathologically positive samples were confirmed by direct fluorescent antibody test and virus isolation. Of the 11 ILT-positive cases, 10 were positive by nested PCR. In addition, ILTV DNA was detected in 7 of the 24 samples diagnosed as NST upon histopathologic examination. Therefore, by nested PCR, ILTV DNA was detected in tissues independently of the presence of syncytial cells, intranuclear inclusions, or both. ILT nested PCR is a specific and sensitive assay capable of detecting ILT at different stages of infection and can be utilized in combination with histopathological examination to accelerate the diagnosis of ILT infection. PMID- 11922351 TI - Effect of antibiotics on in vitro and in vivo avian cartilage degradation. AB - Antibiotics are used in the livestock industry not only to treat disease but also to promote growth and increase feed efficiency in less than ideal sanitary conditions. However, certain antibiotic families utilized in the poultry industry have recently been found to adversely affect bone formation and cartilage metabolism in dogs, rats, and humans. Therefore, the first objective of this study was to determine if certain antibiotics used in the poultry industry would inhibit in vitro cartilage degradation. The second objective was to determine if the antibiotics found to inhibit in vitro cartilage degradation also induced tibial dyschondroplasia in growing broilers. Ten antibiotics were studied by an avian explant culture system that is designed to completely degrade tibiae over 16 days. Lincomycin, tylosin tartrate, gentamicin, erythromycin, and neomycin sulfate did not inhibit degradation at any concentration tested. Doxycycline (200 microg/ml), oxytetracycline (200 microg/ml), enrofloxacin (200 and 400 microg/ml), ceftiofur (400 microg/ml), and salinomycin (10 microg/ml) prevented complete cartilage degradation for up to 30 days in culture. Thus, some of the antibiotics did inhibit cartilage degradation in developing bone. Day-old chicks were then administered the five antibiotics at 25%, 100%, or 400% above their recommended dose levels and raised until 21 days of age. Thiram, a fungicide known to induce experimental tibial dyschondroplasia (TD), was given at 20 ppm. Birds were then killed by cervical dislocation, and each proximal tibiotarsus was visually examined for TD lesions. The results showed that none of these antibiotics significantly induced TD in growing boilers at any concentration tested, whereas birds given 20 ppm thiram had a 92% incidence rate. PMID- 11922352 TI - Growth and infectivity assays of the Israeli vaccine strain of fowl poxvirus in chicken embryo fibroblasts. AB - The Israeli vaccine strain of fowl poxvirus grows efficiently in chicken embryo fibroblasts but not in cell lines derived from monkey kidney or human fibroblasts. We developed two assays for the titration of the infectivity of this virus in secondary cultures of chicken embryo fibroblasts. The first is a focus assay, in which minimum essential medium and SeaKem ME agarose were used for the overlay media. Under these conditions, clear virus foci appeared after 5 days of incubation at 37 C. The second assay is a semiautomatic colorimetric test based on the ability of live cells in culture to reduce the yellow tetrazolium salt 3 [4,5-dimethylthiazol-2-yl]-2,5-diphenyltetrazolium bromide (MTT; thiazolyl blue) to its formazan derivative. The reagent was added to infected chicken embryo fibroblasts in 96-well plates 10 days after infection. The formazan formed during 2 hr was extracted with dimethyl sulfoxide, and its absorbance was read by an automatic microplate spectrophotometer. A good correlation of the infectivity titers of the virus was obtained by the two methods. PMID- 11922353 TI - Commemoration of the centenary of the birth of Sir Reginald Watson-Jones. PMID- 11922354 TI - Paget's disease of the bone and its management. PMID- 11922355 TI - Total knee arthroplasty in patients with haemophilia who are HIV-positive. PMID- 11922356 TI - Fractures of the glenoid treated by operation. A 5- to 23-year follow-up of 22 cases. AB - We describe the results after open reduction and internal fixation of 22 consecutive displaced fractures of the glenoid with a mean follow-up of ten years. A posterior approach was used in 16 patients and an anterior in six, the approach being chosen according to the Ideberg classification of the fractures. The fixation failed in two patients, one of whom required a further operation. There were two cases of deep infection. At follow-up the median Constant score was 94% (mean 79%, range 17 to 100). The score was less than 50% in four patients, including the two who became infected. A further two had an associated complete palsy of the brachial plexus. PMID- 11922357 TI - The role of angiography in the management of haemorrhage from major fractures of the pelvis. AB - In a series of 150 consecutive patients with unstable fractures of the pelvis, angiography was performed in 23 (15%) who had uncontrolled hypotension. There were three anteroposterior compression (APC), eight lateral compression (LC) and 12 vertical shear (VS) injuries. Arterial sources of haemorrhage were identified in 18 (78%) patients and embolisation was performed. Angiography was required in 28% of VS injuries. The morphology of the fracture was not a reliable guide to the associated vascular injury. Ten (43%) patients died, of whom six had had angiography as the first therapeutic intervention. Five of these had a fracture which was associated with an increase in pelvic volume (APC or VS) which could have been stabilised by an external fixator. Based on our findings we recommend skeletal stabilisation and, if indicated, laparotomy to deal with sources of intraperitoneal blood loss before pelvic angiography. Embolisation of pelvic arterial bleeding is a worthwhile procedure in patients with hypotension which is unresponsive to these interventions. PMID- 11922358 TI - A prospective randomised trial of internal fixation versus arthroplasty for displaced fractures of the neck of the femur. Functional outcome for 450 patients at two years. AB - It remains a matter of debate whether displaced fractures of the neck of the femur should be treated by internal fixation or arthroplasty. We have compared the two methods with regard to complications, mortality and functional outcome. We studied 409 patients, aged 70 years and over, with subcapital fractures graded as Garden 3 or 4, in a two-year prospective multicentre study from 12 Swedish hospitals. They were randomised to internal fixation or arthroplasty. Patients who were mentally confused, bedridden or in a nursing-home were excluded from the survey. After two years the rate of failure was 43% in the internal fixation (IF) and 6% in the arthroplasty group (p < 0.001). In the IF group 36% had impaired walking and 6% had severe pain compared with 25% and 1.5%, respectively, in the arthroplasty group (both p < 0.05). There was no difference in mortality. With a high rate of failure and poor functional outcome after IF, we recommend primary arthroplasty for displaced fractures of the neck of the femur in patients over 70 years of age. PMID- 11922359 TI - Somatisation predicts the outcome of treatment in patients with low back pain. AB - We have assessed the influence of somatisation on the outcome of treatment in 81 patients with chronic low back pain. All, irrespective of whether treatment was surgical or conservative, had a significantly better (p < 0.001) health-related quality of life at follow-up on all but one scale of the SF-36. Lower health related quality of life at follow-up correlated significantly with a higher tendency to somatise before treatment and at follow-up. A logistic regression analysis yielded two factors which predicted the outcome; somatisation (p < 0.001) and 'doctor shopping' (the number of physicians consulted before the present inpatient treatment, p < 0.001). These factors accurately distinguished between patients with good and those with poor outcomes in 82%. Patients with somatisation and 'doctor shopping' were at a higher risk for a poor outcome. The results show the relevance of somatisation in the outcome of treatment in patients with low back pain. PMID- 11922360 TI - The position of the tourniquet on the upper limb. AB - Our aim was to determine if a tourniquet placed on the forearm has any advantage in clinical practice over the usual position on the upper arm. We randomised 50 patients who were undergoing an open operation for carpal tunnel syndrome under local anaesthesia into two groups. One had a tourniquet on the upper arm and the other on the forearm. The blood pressure, pulse, and level of pain were recorded at intervals of five minutes during the operation. The surgeons were also asked to evaluate the quality of the anaesthesia, the bloodless field, and the site of the tourniquet. The patients tolerated the tourniquet on the upper arm and forearm equally well. The surgeons had some difficulties when it was placed on the forearm. We therefore recommend placement of a tourniquet on the upper arm for operations on the hand and wrist which are carried out under local anaesthesia. PMID- 11922361 TI - Distraction lengthening by callotasis in the hand. AB - The clinical results of 26 digits (18 patients) lengthened by distraction callotasis were evaluated and the factors which influenced healing were analysed. There were 14 men and four women, with a mean age of 39 years. All digits had suffered traumatic amputation. There were eight thumbs and 18 fingers. The level of the site of the osteotomy was at the proximal metaphysis in ten, the middle diaphysis in 13 and the distal metaphysis in three. Although the proposed length was achieved in 23 of the 26 digits, five required additional bone grafts. The rate of healing was 96 days/cm in the digits without complications such as callus fracture or poor callus formation, and 158 days/cm in those with complications. Lengthening at the proximal metaphysis gave a better result than at the diaphysis or distal metaphysis. PMID- 11922362 TI - Total knee arthroplasty in neuropathic arthropathy. AB - We describe the results of total knee arthroplasty (TKA) undertaken for severe, neurosyphilitic Charcot arthropathy in ten patients (19 knees). A cemented condylar, constrained prosthesis was implanted in all but two knees. The mean follow-up was 5.2 years (5 to 6). The mean knee score before operation was 36.5 points (30 to 42) which improved to 76 points (58 to 90) after operation as judged by the Hospital for Special Surgery score. At final follow-up three knees (16%) had aseptic loosening which required salvage by an arthrodesis, six (31%) were functioning poorly and ten (53%) were satisfactory. We conclude that although Charcot arthropathy is not an absolute contraindication to total knee replacement, there is a high incidence of serious complications. PMID- 11922363 TI - Ability to kneel after total knee replacement. AB - We have investigated the ability to kneel after total knee replacement. We asked 75 patients (100 knees) at least six months after routine uncemented primary total knee replacement, to comment on and to demonstrate their ability to kneel. Differences between the perceived and actual ability to kneel were noted. In 32 knees patients stated that they could kneel without significant discomfort. In 54 knees patients avoided kneeling because of uncertainties or recommendations from third parties (doctors, nursing staff, friends, etc). A total of 64 patients was actually able to kneel without discomfort or with mild discomfort only and 12 of the remainder were unable to kneel because of problems which were not related to the knee. Twenty-four patients therefore were unable to kneel because of discomfort in the knee. There was no difference between the 'kneelers' and 'non kneelers' with regard to overall knee score, range of movement and the presence of patellar resurfacing. PMID- 11922364 TI - Varus inclination of the distal femur and high tibial osteotomy. AB - We have analysed retrospectively the relationship between the axial parameters of alignment of the lower limb and the recurrence of varus deformity after high tibial osteotomy. We studied 29 patients (37 knees) with a mean age at surgery of 66 years. The mean follow-up was for 7.4 years (5 to 10.5). Recurrence of varus deformity was defined as an increase in the femorotibial angle of 3 degrees or more, compared with that obtained six months after the operation. There were four patients (four knees) with recurrence of varus deformity. They had a greater varus inclination of the distal femur than those without varus recurrence. An association between varus inclination of the distal femur and horizontal obliquity of the joint surface was observed. Excessive obliquity prevents the shift of weight-bearing to the lateral compartment, and may cause a recurrence of varus deformity after high tibial osteotomy. PMID- 11922365 TI - Treatment of cartilage defects of the talus by autologous osteochondral grafts. AB - We reviewed retrospectively 11 patients who had been treated surgically by open autologous osteochondral grafting for symptomatic chondral or osteochondral defects of the dome of the talus between 1996 and 1999. The mean ages of the eight men and three women were 34.2 and 25.9 years, respectively, with a mean time to follow-up of 24 months. The results of functional outcome were prospectively obtained using the MODEMS AAOS foot and ankle follow-up questionnaire, the AOFAS ankle-hindfoot scale and the Hannover scores for the ankle. The grafts were harvested from the ipsilateral knee. Good to excellent results were obtained for the ankle without adverse effects on the knee. We believe that autologous osteochondral grafting should be considered for the patient with a symptomatic osteochondral defect of the talus. PMID- 11922366 TI - Cannulation of simple bone cysts. AB - We describe a consecutive series of 26 patients with simple bone cysts who were treated by curettage, multiple drilling and continuous decompression by the insertion of either a cannulated screw or a pin. In the first 15 patients we used titanium cannulated screws (group 1) and in the next 11 a cannulated hydroxyapatite pin (group 2). Satisfactory healing was achieved in 12 patients in group 1 (80%) and in all in group 2. This technique seems to be a promising option for the treatment of simple bone cysts. The cannulated hydroxyapatite pin is recommended because of its higher success rate and the fact that it does not need to be removed. PMID- 11922367 TI - Peripheral osteoid osteoma. Is there still a place for traditional surgery? AB - We treated 106 patients with a peripheral osteoid osteoma by conventional surgical methods; 81 had curettage and 25 en-bloc resection. The rate of local recurrence after curettage was 12% and after en-bloc resection 4.5%. Postoperative fractures were observed in 3% after curettage and in 4.5% after en bloc resection. We compared our findings with those reported in the literature after minimally invasive treatment and concluded that curettage can be regarded as the treatment of choice in patients in whom minimally invasive methods do not offer any advantage, for example, for subperiosteal tumours which are readily accessible, or when the diagnosis is unclear and further histological analysis is required. PMID- 11922368 TI - Congenital insensitivity to pain. Orthopaedic manifestations. AB - We reviewed 13 patients with congenital insensitivity to pain. A quantitative sweat test was carried out in five and an intradermal histamine test in ten. DNA examination showed specific mutations in four patients. There were three clinical presentations: type A, in which multiple infections occurred (five patients); type B, with fractures, growth disturbances and avascular necrosis (three patients); and type C, with Charcot arthropathies and joint dislocations, as well as fractures and infections (five patients, four with mental retardation). Patient education, shoeware and periods of non-weight-bearing are important in the prevention and early treatment of decubitus ulcers. The differentiation between fractures and infections should be based on aspiration and cultures to prevent unnecessary surgery. Established infections should be treated by wide surgical debridement. Deformities can be managed by corrective osteotomies, and shortening by shoe raises or epiphysiodesis. Joint dislocations are best treated conservatively. PMID- 11922369 TI - Osteochondritis dissecans of the knee in children. A comparison of MRI and arthroscopic findings. AB - The treatment of osteochondritis dissecans (OCD) in children and adolescents is determined by the stability of the lesion and the state of the overlying cartilage. MRI has been advocated as an accurate way of assessing and staging such lesions. Our aim was to determine if MRI scans accurately predicted the subsequent arthroscopic findings in adolescents with OCD of the knee. Some authors have suggested that a high signal line behind a fragment on the T2 weighted image indicates the presence of synovial fluid and is a sign of an unstable lesion. More recent reports have suggested that this high signal line is due to the presence of vascular granulation tissue and may represent a healing reaction. We were able to improve the accuracy of MRI for staging the OCD lesion from 45% to 85% by interpreting the high signal T2 line as a predictor of instability only when it was accompanied by a breach in the cartilage on the T1 weighted image. We conclude that MRI can be used to stage OCD lesions accurately and that a high signal line behind the OCD fragment does not always indicate instability. We recommend the use of an MRI classification system which correlates with the arthroscopic findings. PMID- 11922370 TI - Natural history of infantile tibia vara. AB - We have observed the natural history without treatment of 46 limbs in 29 patients with infantile tibia vara and a metaphyseal-diaphyseal angle (MDA) of more than 11 degrees. The femorotibial angle (FTA) and MDA were measured, and Langenskiold's classification of radiological changes in the proximal medial metaphysis of the tibia was used. In 22 limbs which were not in Langenskiold stages II to III the condition resolved spontaneously without treatment. Of the remaining 24 which were in stages II to III, in 18 it resolved spontaneously by the age of six years, but six showed little or no improvement at the latest follow-up. It was impossible to differentiate by measuring the FTA or MDA whether spontaneous resolution could be expected before the age of four years. There was no difference in the rate of resolution of the deformity between those patients who had been treated by a brace and those who had received no treatment. We advise no initial treatment but review at six-monthly intervals until the age of four years, even in patients with Langenskiold stage-II to stage-III deformity. When a deformity persisted or progressed we carried out a corrective osteotomy after the age of four years. PMID- 11922371 TI - A simple guide to determine the magnification of radiographs and to improve the accuracy of preoperative templating. AB - Templates are used in the preoperative planning of many orthopaedic procedures. The magnification of the bones on preoperative radiographs can vary despite using standardised radiological techniques. Templates will give misleading measurements unless this magnification is quantified. A coin may be used to calculate the magnification, with significant improvement in the accuracy of templating (p = 0.05). A group of patients undergoing uncemented arthroplasty of the hip was at high risk for intraoperative fracture of the femur because the magnification of the radiograph was larger than that of the template. PMID- 11922372 TI - Methicillin-resistant Staphylococcus aureus osteomyelitis of the scaphoid from a catheter in the radial artery. AB - We describe the development of methicillin-resistant Staphylococcus aureus osteomyelitis of the scaphoid in a 49-year-old man from an infection occurring around a catheter in the radial artery. Total scaphoidectomy and appropriate antibiotic therapy eradicated the infection. PMID- 11922373 TI - Insulin-like growth factor-I enhances cell-based repair of articular cartilage. AB - Composites of chondrocytes and polymerised fibrin were supplemented with insulin like growth factor-I (IGF-I) during the arthroscopic repair of full-thickness cartilage defects in a model of extensive loss of cartilage in horses. Repairs facilitated with IGF-I and chondrocyte-fibrin composites, or control defects treated with chondrocyte-fibrin composites alone, were compared before death by the clinical appearance and repeated analysis of synovial fluid, and at termination eight months after surgery by tissue morphology, collagen typing, and biochemical assays. The structure of cartilage was evaluated histologically by Toluidine Blue reaction and collagen type-I and type-II in situ hybridisation and immunohistochemistry. Repair tissue was biochemically evaluated by DNA assay, proteoglycan quantitation and characterisation, assessment of collagen by reverse phase high-performance liquid chromatography, and collagen typing using cyanogen bromide digestion and peptide separation by polyacrylamide gel electrophoresis. The results at eight months showed that the addition of IGF-I to chondrocyte grafts enhanced chondrogenesis in cartilage defects, including incorporation into surrounding cartilage. Gross filling of defects was improved, and the tissue contained a higher proportion of cells producing type-II collagen. Measurements of collagen type II showed improved levels in IGF-I-treated defects, supporting in situ hybridisation and immunohistochemical assessments of the defects. IGF-I improves the repair capabilities of chondrocyte-fibrin grafts in large full thickness repair models. PMID- 11922374 TI - Intervertebral disc degeneration adjacent to a lumbar fusion. An experimental rabbit model. AB - Our study establishes a rabbit model of disc degeneration which requires neither a chemical nor physical injury to the disc. Disc degeneration similar to that seen in man was created at levels proximal (L4-L5) and caudal (L7-S1) to a simulated lumbar fusion and was studied for up to nine months after arthrodesis. Loss of the normal parallel arrangement of collagen bundles within the annular lamellae was observed in intervertebral discs adjacent to the fusion at three months. By six months there was further disorganisation as well as loss of distinction between the lamellae themselves. By nine months the structure of the disc had been replaced by disorganised fibrous tissue, and annular tears were seen. There was an initial cellular proliferative response followed by loss of chondrocytes and notochordal cells in the nucleus pulposus. Degeneration was accompanied by a decrease in the monomer size of proteoglycans. Narrowing of the disc space, endplate sclerosis and the formation of osteophytes at adjacent disc spaces were observed radiologically. PMID- 11922375 TI - Endotoxin contamination of particles produces misleading inflammatory cytokine responses from macrophages in vitro. AB - Particulate prosthetic materials are often studied by adding them to monocytic cells in vitro and measuring the release of cytokines as an indicator of their inflammatory potential. Endotoxin is known to be a contaminant of particle preparations and also stimulates the release of cytokines. It is usual to use a proprietary endotoxin test to avoid erroneous results. Four different formulations of cement were found to be free from endotoxin using standard, gel clot tests but stimulated different levels of release of cytokines from macrophages. These differences were explained when a more sensitive, kinetic endotoxin assay showed that release correlated with minor contamination with endotoxin. In a repeat experiment using cement particles with low or undetectable levels of endotoxin by kinetic assay, differences in the ability of the formulations to stimulate the release of cytokines were not seen. Endotoxin is adsorbed on to the surface of particles and it is this combination which stimulates increased release of cytokines. In both the above methods for determination of endotoxin, the water in which the particles had been soaked was examined rather than the particles directly. Further investigations showed that a kinetic assay directly on a particle suspension is the most sensitive method to measure contamination with endotoxin. PMID- 11922376 TI - Perfusion of the femoral head during surgical dislocation of the hip. Monitoring by laser Doppler flowmetry. AB - We used laser Doppler flowmetry (LDF) with a high energy (20 mW) laser to measure perfusion of the femoral head intraoperatively in 32 hips. The surgical procedure was joint debridement requiring dislocation or subluxation of the hip. The laser probe was placed within the anterosuperior quadrant of the femoral head. Blood flow was monitored in specific positions of the hip before and after dislocation or subluxation. With the femoral head reduced, external rotation, both in extension and flexion, caused a reduction of blood flow. During subluxation or dislocation, it was impaired when the posterosuperior femoral neck was allowed to rest on the posterior acetabular rim. A pulsatile signal returned when the hip was reduced, or was taken out of extreme positions when dislocated. After the final reduction, the signal amplitudes were first slightly lower (12%) compared with the initial value but tended to be restored to the initial levels within 30 minutes. Most of the changes in the signal can be explained by compromise of the extraosseous branches of the medial femoral circumflex artery and are reversible. Our study shows that LDF provides proof for the clinical observation that perfusion of the femoral head is maintained after dislocation if specific surgical precautions are followed. PMID- 11922377 TI - Radiation therapy for the prevention of heterotopic ossification at the elbow. PMID- 11922378 TI - Excision of hemivertebrae in the management of congenital scoliosis. PMID- 11922379 TI - The Lubinus patellofemoral arthroplasty: a five- to ten-year prospective study. PMID- 11922380 TI - The Lubinus patellofemoral arthroplasty: a five- to ten-year prospective study. PMID- 11922381 TI - A prospective comparative analysis of mobility in osteoarthritic knees: does lifestyle have an influence? PMID- 11922382 TI - Screening for amblyopia in preschool children: results of a population-based, randomised controlled trial. ALSPAC Study Team. Avon Longitudinal Study of Pregnancy and Childhood. AB - INTRODUCTION: The rationale for preschool vision screening programmes has recently been questioned. Evidence about the effects of early treatment is needed, but it is not known how early the target conditions can reliably be detected. In this study, an intensive programme comprising several different screening methods, used at different ages up to 37 months, was compared with the usual practice of visual surveillance and ad hoc referrals. METHODS: Two groups were randomly selected from children in a population birth cohort study. The control group (n = 1461) received visual surveillance only. The intervention group (n = 2029) was offered in addition a programme of regular visual assessments by orthoptists testing visual acuity, ocular alignment, stereopsis and non-cycloplegic photorefraction. RESULTS: The intervention group programme yielded more children with amblyopia (1.6% vs. 0.5%, p < 0.01), and was more specific (95% vs. 92%, p < 0.01), than the control programme. The individual components of the intervention programme were compared. The cover test and visual acuity tests were poorly sensitive until the children were 37 months, but were always >99% specific. Photorefraction was more sensitive than acuity testing at all ages below 37 months, with specificity >95% at 31 and 37 months. CONCLUSIONS: Photorefraction would have detected more children less than 37 months of age with straight-eyed amblyopia than did visual acuity testing, but with more false positives. At 37 months, photorefraction plus a cover test would have been comparable in effectiveness to visual acuity testing plus a cover test. PMID- 11922384 TI - Incidence of and risk factors for diabetic retinopathy in diabetic clinic attenders. AB - The incidence and risk factors for the development of diabetic retinopathy during a mean (SD) follow-up period of 4.6 (2.9) (range 1-12.4) years have been examined among 3424 patients (1878 males and 1546 females) with diabetes mellitus from three outpatient clinics at the University Hospital, Nottingham. The mean (SD) age of participants was 49.2 (17.9) years with a mean (SD) duration of diabetes of 7.3 (9.0) years at initial registration. Among the 3424 patients free of retinopathy at initial registration who attended the clinic at least twice in the period 1979-1992, the incidence of any retinopathy was 59.6 (57.8 male and 61.8 female) per 1000 person-years based on 15,571 person-years of follow-up. The incidence rate of retinopathy was 72% higher among insulin-treated than among non insulin-treated noninsulin-dependent diabetes mellitus (NIDDM) clinic attenders. Using a Cox's Proportional Hazards Model for insulin-dependent diabetes (IDDM) and NIDDM (insulin and non-insulin-treated) diabetes separately, longer duration of diabetes, higher systolic blood pressure and poor metabolic control were significant independent predictors of retinopathy for all three groups. Never smoking was a significant independent predictor of retinopathy for the insulin dependent diabetes groups. Lower body mass index, proteinuria and age were predictors of retinopathy only for non-insulin-treated NIDDM patients. Gender and creatinine had no significant independent association with retinopathy when other covariates were considered. These findings will help the identification of those patients at particular risk of retinopathy so that clinic time for screening of eyes can be appropriately focused. PMID- 11922383 TI - Evaluating a visual field screening test for glaucoma: how the choice of the gold standard affects the validity of the test. AB - OBJECTIVE: To examine the effects of the gold standard on sensitivity, specificity, and area under the ROC curve when determining the validity of a screening test. METHODS: Fifty-eight people were recruited from a Veterans' Administration eye clinic. Screening included testing with the 26-point oculokinetic perimeter (OKP). We used different gold standards to categorize patients as positive or negative for glaucoma. The glaucoma hemifield test (GHT) and corrected pattern standard deviation (CPSD) from the Humphrey visual field analyzer (HVF) were used as objective gold standards. Ophthalmologist review of the HVF, using two different methods for classifying visual field defects, was also used as a gold standard. RESULTS: Using the review of the HVF that included mild, moderate, or severe defects, the area under the ROC curve was not significantly different from 50%. However, for all the other gold standards, the area under the ROC curve was significantly different from 50%. CONCLUSIONS: In determining the ability of the OKP to differentiate between glaucoma and normal subjects, the choice of a gold standard led to different conclusions about its usefulness as a screening test for glaucoma. Furthermore, sensitivity and specificity varied widely depending on which gold standard was used, and what cutoff point was used for the OKP. Although clinician review of the HVF may be used when specific criteria are given to define visual field defects, using objective measures from the HVF is easier and will allow for better comparisons between studies. PMID- 11922385 TI - Reliability and reproducibility of a Chinese-language visual function assessment. AB - OBJECTIVE: The purpose of the study was to validate a Chinese-language visual function assessment within the context of a routine cataract surgery practice and to assess the contribution of the method of questionnaire administration. DESIGN: The visual function assessment (VFA) was translated into Chinese. Two groups of study subjects were recruited: Chinese who did not speak English and Chinese conversant in English. Consecutive preoperative cataract patients of Chinese ancestry presenting to an urban ophthalmology practice were recruited. The questionnaire was administered in person or by telephone interview. Pre-operative visual acuity was recorded. Visual function scores were analyzed to assess reliability and correlation with visual acuity. RESULTS: Among the 186 potential study subjects, 155 patients completed the study The Chinese-language visual function assessment had good internal consistency (Cronbach alpha = 0.97, inter item correlations = 0.43 to 0.96) . Reliability (with regard to the English version) and test-retest reproducibility of the Chinese questionnaire were strong with intraclass correlation coefficients greater than 0.60. The method of administration contributed to the measures of reliability and reproducibility. CONCLUSION: These results show that a Chinese-language version of the VFA questionnaire is reliable and valid. In industrialized countries with large Chinese-speaking populations and newly developed countries of East and Southeast Asia, the visual function assessment may be helpful in assisting routine clinical patient evaluation and cross-cultural outcome assessment programmes. Our findings also suggest that self-administered visual function assessments may be more reliable and valid than interview-generated assessments. PMID- 11922386 TI - Visual impairment in a Taiwanese population: prevalence, causes, and socioeconomic factors. AB - PURPOSE: We sought to estimate the prevalence of visual impairment among an elderly population in urban Taiwan, determine the causes of visual impairment, and gain information about certain socioeconomic factors associated with visual impairment. METHODS: A population-based survey of ocular diseases in residents aged 50 years or older was conducted in Peitou Precinct, Taipei, Taiwan. All participants underwent a comprehensive ophthalmic examination, including visual acuity measurement, using standardized protocols. Demographic data, marital status, employment status, and educational level were gathered through an interview. RESULTS: A total of 2034 participants completed the visual acuity measurements. The prevalence of visual impairment (defined as best-corrected visual acuity in the better eye <6/18) among this population was 2.75% (95% confidence intervals: 2.04%-3.46%). Three major causes of visual impairment were cataract (30.4%), high myopic macular degeneration (25.0%), and age-related macular degeneration (14.3%). In multivariate analysis, age was positively correlated with visual impairment (P < 0.001), and higher education level was associated with a significant decrease in the odds of being visually impaired (P < 0.001). No relation was found between visual impairment and sex, or marital and employment status. CONCLUSIONS This study provides the first information about the prevalence and causes of visual impairment in Taiwan, and identifies age and educational level as the most important factors related to visual impairment. Additional educational programs should be developed to improve individual awareness of age-related ocular diseases and the availability of current ophthalmic intervention. PMID- 11922387 TI - Stable karyotypes in epithelial cancer cell lines despite high rates of ongoing structural and numerical chromosomal instability. AB - Most human tumors and tumor cell lines exhibit numerical and structural chromosomal abnormalities. The goal of this study was to determine the ongoing rates of structural and numerical instability in selected cancer cell lines and to investigate the consequences of these rates to karyotypic progression. We studied two colorectal (HCT-116 and HT-29) and two ovarian (SKOV-3 and OVCAR-8) cancer cell lines and their single cell subclones. We found that the signature karyotypes of all four cell lines were distinct and each aberrant. Whereas high rates of ongoing structural and/ or numerical chromosomal instability could be demonstrated in all cell lines, there was a relative stability of the consensus karyotype over many generations. No new clonal structural chromosomal reconfigurations emerged and the few numerical changes of karyotypes were restricted to abnormal chromosomes. This implies a kind of genomic optimization under the conditions of cell culture and suggests a link between genomic stabilization and cell propagation. We have been able to support this possibility by computer modeling. We did not observe a profound difference in the rates of numerical or structural instability in the cell lines with a replication error phenotype (RER+) versus the other cell lines. PMID- 11922388 TI - Systemic distribution and tumor localization of adoptively transferred lymphocytes in mice: comparison with physiologically based pharmacokinetic model. AB - The mechanisms by which tumors are able to evade cellular immune responses are still largely unknown. It is likely, however, that the initial recruitment of lymphocytes to tumor vessels is limited by cell retention in normal tissue, which results in a low flux of these cells into the tumor vasculature. We grew MCaIV (mouse mammary carcinoma) tumors in the leg of SCID mice and injected 111In-oxine labeled, primed T lymphocytes directed against the tumor intravenously. The systemic distribution of cells in normal organs was similar between mice injected with primed and control lymphocyte populations, except for a delayed clearance of primed lymphocytes from the lungs. Kinetics of lymphocyte localization to the tumor were identical between the primed and control lymphocyte populations. Splenectomy before the injection of primed lymphocytes increased delivery of cells to the lungs and liver after 1 hour with no significant improvement in tumor localization. Within 24 to 168 hours after injection, localization of cells in the liver of splenectomized mice was higher than in the control group. However, no significant difference in tumor localization was observed between groups. A physiologically based compartmental model of lymphocyte distribution predicted the compartmental sequestration and identified model parameters critical for experimental planning and therapeutic optimization. PMID- 11922389 TI - MYC expression promotes the proliferation of neural progenitor cells in culture and in vivo. AB - Primitive neuroectodermal tumors (PNETs) are pediatric brain tumors that result from defects in signaling molecules governing the growth and differentiation of neural progenitor cells. We used the RCAS-TVA system to study the growth effects of three genetic alterations implicated in human PNETs on a subset of neural progenitor cells that express the intermediate filament protein, nestin. The genetic alterations tested were: 1) overexpression of the cellular oncoprotein, MYC; 2) activation of transcription factor, beta-catenin; and 3) haploinsufficiency of Ptc, the hedgehog receptor gene. The RCAS-TVA system uses an avian retroviral vector, RCAS, to target gene expression to specific cell types in transgenic mice. To express exogenous genes in neural progenitor cells, we used Ntv-a mice. In these mice, the Nestin gene promoter drives expression of TVA, the cell surface receptor for the virus. Ectopic expression of MYC, but not activated beta-catenin, promoted the proliferation of neural progenitor cells in culture and in the cerebral leptomeninges in vivo. These effects were equally penetrant in mice with Ptc+/- and Ptc+/+ genetic backgrounds. Although overexpression of MYC is not sufficient to cause intraparenchymal tumors, it may facilitate PNET formation by sustaining the growth of undifferentiated progenitor cells. PMID- 11922391 TI - Cytotoxic cyplasin of the sea hare, Aaplysia punctata, cDNA cloning, and expression of bioactive recombinants in insect cells. AB - A 56-kDa protein isolated from the mucus of the European sea hare Aplysia punctata shows a prefer ential toxicity to autonomously growing transformed mammalian cells. Cell death induced by this protein differs from both apoptosis and necrosis. The cytotoxic effects are irreversible and become apparent at nanomolar concentrations in a cell type-dependent manner. In contrast, injection of micromolar concentrations into mice is tolerated without apparent negative consequences. Microsequencing of the 56-kDa protein released a peptide sequence whose corresponding nucleotide sequence was used as probe to screen A. punctata RNA-based cDNA and to select cDNA clones encoding polypeptides comprising the target peptide. Two closely related cDNA were detected. The cDNA encoding a polypeptide 558 aa in length was considered to reflect a bonafide clone encoding the cytotoxic protein. Its protein-coding section was recloned in vectors suitable for expression in Escherichia coli, in mammalian cells, and in insect cells, respectively. The E. coli-expressed polypeptide was biologically inactive. Transfected mammalian cells expressed a cytotoxic factor and died thereof as if treated with the genuine cytotoxic protein. In contrast, transfected insect cells, which proved to be much less sensitive when treated with the genuine protein, expressed the cytotoxic factor and continued to proliferate, allowing to establish stable insect cell lines expressing sufficient amounts of the cytotoxic factor for further characterization. PMID- 11922390 TI - Hypoxia-inducible regulation of a prodrug-activating enzyme for tumor-specific gene therapy. AB - Previous studies have suggested that tumor hypoxia could be exploited for cancer gene therapy. Using hypoxia-responsive elements derived from the human vascular endothelial growth factor gene, we have generated vectors expressing a bacterial nitroreductase (NTR) gene that can activate the anticancer prodrug CB1954. Stable transfectants of human HT1080 tumor cells with hypoxia-inducible vectors were established with G418 selection. Hypoxic induction of NTR protein correlated with increased sensitivity to in vitro exposure of HT1080 cells to the prodrug. Growth delay assays were performed with established tumor xenografts derived from the same cells to detect the in vivo efficacy of CB1954 conversion to its cytotoxic form. Significant antitumor effects were achieved with intraperitoneal injections of CB1954 both in tumors that express NTR constitutively or with a hypoxia inducible promoter. In addition, respiration of 10% 02 increased tumor hypoxia in vivo and enhanced the antitumor effects. Taken together, these results demonstrate that hypoxia-inducible vectors may be useful for tumor-selective gene therapy, although the problem of delivery of the vector to the tumors, particularly to the hypoxic cells in the tumors, is not addressed by these studies. PMID- 11922392 TI - Fibroblast growth factor-1 induced promatrilysin expression through the activation of extracellular-regulated kinases and STAT3. AB - The MMP, matrilysin (MMP-7), has been shown to be overexpressed in prostate cancer cells and to increase prostate cancer cell invasion. Prostate stromal fibroblasts secrete factor(s), including fibroblast growth factor-1 (FGF-1) that induces promatrilysin expression in LNCaP cells. In the present study, we investigated the signal transduction pathway involved in the FGF-1-induced expression of promatrilysin. FGF-1 treatment significantly increased the activation of extracellular signal-regulated kinases 1 and 2 (ERK1 and ERK2). This induction was time-dependent and was sustained until 24 hours after treatment. Treating the cells with MEK1/2 inhibitor (PD98059) eliminated ERK activation completely and blocked FGF-1-mediated induction of promatrilysin expression. Transient transfection studies with human matrilysin promoter resulted in a four- to five-fold increase in reporter luciferase enzyme activity that was blocked by the MEK1/2 inhibitor (PD98059). Serine phosphorylation of signal transducer and activator of transcription 3 (STAT3) was observed after FGF 1 treatment and pretreatment with 20 microM PD98059-abolished STAT3 phosphorylation. Transient transfection with dominant negative STAT3 inhibited FGF-1-induced transactivation of the matrilysin promoter indicating that STAT3 plays an important role in FGF1-induced matrilysin expression. We propose that the FGF-1-induced signaling pathway that leads to promatrilysin expression is ERK dependent and leads to phosphorylation of Ser-727 on STAT3, phosphorylated STAT3, then binds and transactivates the matrilysin promoter. Our results demonstrate that ERK-MAP kinase and transcription factor STAT3 are important components of FGF-1-mediated signaling, which induce promatrilysin expression in LNCaP cells. PMID- 11922393 TI - Glucocorticoids and polyamine inhibitors synergize to kill human leukemic CEM cells. AB - Glucocorticoids are well-known apoptotic agents in certain classes of lymphoid cell malignancies. Reduction of intracellular polyamine levels by use of inhibitors that block polyamine synthesis slows or inhibits growth of many cells in vitro. Several such inhibitors have shown efficacy in clinical trials, though the toxicity of some compounds has limited their usefulness. We have tested the effects of combinations of the glucocorticoid dexamethasone (Dex) and two polyamine inhibitors, difluoromethylornithine (DFMO) and methyl glyoxal bis guanylhydrazone (MGBG), on the clonal line of human acute lymphoblastic leukemia cells, CEM-C7-14. Dex alone kills these cells, though only after a delay of at least 24 hours. We also evaluated a partially glucocorticoid-resistant c-Myc expressing CEM-C7-14 clone. We show that Dex downregulates ornithine decarboxylase (ODC), the rate-limiting enzyme in polyamine synthesis. Pretreatment with the ODC inhibitor DFMO, followed by addition of Dex, enhances steroid-evoked kill slightly. The combination of pretreatment with sublethal concentrations of both DFMO and the inhibitor of S-adenosylmethionine decarboxylase, MGBG, followed by addition of Dex, results in strong synergistic cell kill. Both the rapidity and extent of cell kill are enhanced compared to the effects of Dex alone. These results suggest that use of such combinations in vivo may result in apoptosis of malignant cells with lower overall toxicity. PMID- 11922394 TI - Identification of semaphorin3B as a direct target of p53. AB - A cDNA microarray analysis indicated that Semaphorin3B (Sema3B), a gene whose product is involved in axon guidance and axonal repulsion, is inducible by p53. Introduction of exogenous p53 into a glioblastoma cell line lacking wild-type p53 (U373MG) dramatically induced expression of Sema3B mRNA. An electrophoretic mobility shift assay and a reporter assay confirmed that a potential p53 binding site present in the promoter region had p53-dependent transcriptional activity. Expression of endogenous Sema3B was induced in response to genotoxic stresses caused by adriamycin treatment or UV irradiation in a p53-dependent manner. Ectopic expression of Sema3B in p53-defective cells reduced the number of colonies in colony formation assays. These results suggest that Sema3B might play some role in regulating cell growth as a mediator of p53 tumor-suppressor activity. PMID- 11922396 TI - Efficacy of omalizumab, an anti-immunoglobulin E antibody, in patients with allergic asthma at high risk of serious asthma-related morbidity and mortality. AB - AIM: Add-on therapy with omalizumab, an anti-immunoglobulin E antibody, is effective in improving disease control in patients with allergic asthma of varying severity. The aim of the present study was to determine the efficacy of omalizumab in a subgroup of patients at high risk of serious asthma-related morbidity and mortality. METHODS: A meta-analysis was performed of three randomised, double-blind, placebo-controlled studies (studies 1, 2 and 3) that enrolled 1412 patients with moderate or severe allergic asthma, all requiring daily treatment with inhaled corticosteroids (ICS). Omalizumab was administered subcutaneously every 2 or 4 weeks at a total 4-weekly dose of at least 0.016 mg/kg/IgE [IU/ml]. Each study consisted of a 16-week steroid-stable phase and a 12-16-week steroid-reduction phase, followed by a 24-week extension phase (studies 1 and 2 only). The primary outcome measure was the annualised rate of significant asthma exacerbation episodes (sAEEs) during the steroid-stable phase for the pooled subgroup of 254 high-risk patients (omalizumab, n = 135; placebo, n = 119). sAEEs were those requiring a doubling of baseline ICS dose (studies 1 and 2 only) or use of systemic steroids (all three studies). RESULTS: Overall, the number of patients with at least one sAEE during the steroid-stable phase was reduced from 35% (42/119) with placebo to 18% (24/135) with omalizumab. Mean sAEE rates were 1.56 and 0.69 per patient-year, respectively, a reduction of 56% with omalizumab (p = 0.007). Similar reductions in exacerbations in favour of omalizumab were observed for the whole study period and for all AEEs. In those with a history of hospitalisation in the last year, 6/49 (12%) on placebo vs. 2/44 (4.5%) on omalizumab were re-hospitalised during the study period. Patients treated with omalizumab also showed significantly greater improvements from baseline in PEFR (p = 0.026), overall AQoL (p = 0.042) and mean nocturnal (p = 0.007) and mean total (p = 0.011) asthma symptom scores compared with placebo. CONCLUSIONS: In patients at high risk of serious asthma-related morbidity and mortality, treatment with omalizumab offers the potential to halve the rate of asthma exacerbations and improve disease control. PMID- 11922397 TI - The acute and sub-chronic effects of levocetirizine, cetirizine, loratadine, promethazine and placebo on cognitive function, psychomotor performance, and weal and flare. AB - AIM: To compare the central and peripheral H1 inhibitory effects of acute and sub chronic doses of levocetirizine (L-CTZ), cetirizine (CTZ), loratadine (LOR) and promethazine (PRM) versus placebo, using a battery of psychomotor and cognitive tests together with measures of the weal and flare reaction. PRM was included in the study as a positive internal control to validate the sensitivity of the psychometric test battery to the CNS effects of the various treatments. METHODS: Twenty healthy volunteers (18-50 years) received L-CTZ 5mg, CTZ 10 mg, LOR 10 mg, PRM 30 mg and placebo once daily for four days in a five-way, double-blind, crossover study. For each treatment condition, subjects were assessed using a psychometric test system and a pinprick weal and flare response to 100 mg/ml histamine solution at baseline and at 1, 2, 3 ,4, 6, 8, 10 and 122 hours post dose on days 1 and 4. The psychometrics comprised critical flicker fusion (CFF), choice reaction time (CRT), a continuous tracking task (CTT) and subjective rating scales for sedation (LARS). On days 2 and 3, subjects took their medication at pre-designated times while out of the unit. RESULTS: The verum (PRM) established the sensitivity of the test battery: a significant overall reduction in CFF thresholds on both days 1 and 4 (p < 0.05); an overall significant increase (impairment) in recognition, motor and total reaction times on day 1 (p < 0.05); a significant impairment of both the tracking accuracy and reaction time aspects of the CTT task on day 1 (p < 0.005) and significantly higher ratings of subjective sedation on day 1 (p < 0.05). L-CTZ, CTZ and LOR were not distinguishable from placebo in any of the objective and subjective tests at any time point on either day 1 or day 4. With regards to the peripheral inhibitory effects, L-CTZ inhibited both the weal and flare reaction, with maximum inhibition (almost 100%) occurring within two hours of drug ingestion. CTZ also showed evidence of potent peripheral inhibition of histamine, whereas PRM, and especially LOR, showed only a weak weal and flare reaction which had completely attenuated at day 4. CONCLUSIONS: In a study where the psychometric assessments were shown to be sensitive to impairment, L-CTZ 5 mg was found following both initial and repeated doses, but also to be demonstrably free from disruptive and sedative effects on objective measures of psychomotor and cognitive function. Similarly, CTZ showed evidence of pronounced antihistaminic activity and significantly reduced weal and flare scores after both acute and repeated doses, again without evidence of cognitive or psychomotor impairment. LOR also was non-sedative but the antihistaminic reaction was demonstrably weak. PMID- 11922395 TI - Androgen-dependent regulation of human MUC1 mucin expression. AB - MUC1 mucin is transcriptionally regulated by estrogen, progesterone, and glucocorticoids. Our objective was to determine whether androgen receptor (AR) activation regulates expression of MUC1. The following breast and prostatic cell lines were phenotyped and grouped according to AR and MUC1 protein expression: 1) AR+MUC1 + [DAR17+19 (AR transfectants of DU-145), ZR-75-1, MDA-MB-453, and T47D]; 2) AR-MUC1 + [DZeo1 (AR-vector control), DU-145, BT20, MDA-MB-231, and MCF7]; 3) AR+MUC1 - (LNCaP and LNCaP-r). Cell proliferation was determined using the MTT assay in the presence of synthetic androgen R1881, 0.1 microM to 1 microM. Cell surface MUC1 expression was determined by flow cytometry in the presence or absence of oestradiol, medroxy progesterone acetate or R1881, with and without 4 hydroxy-flutamide (4-OH), a nonsteroidal AR antagonist. The functional significance of MUC1 expression was investigated with a cell-cell aggregation assay. Only AR+MUC1 + cell lines showed a significant increase in MUC1 expression with AR activation (P (range) =.01 to .0001), reversed in the presence of 4-OHF. Cell proliferation was unaffected. Increased expression of MUC1 was associated with a significant (P (range) = .002 to .001) reduction in cell-cell adhesion. To our knowledge, this is the first description of androgen-dependent regulation of MUC1 mucin. This is also functionally associated with decreased cell-cell adhesion, a recognised feature of progressive malignancy. These findings have important implications for physiological and pathological processes. PMID- 11922398 TI - Parenteral troxerutin and carbazochrome combination in the treatment of post hemorrhoidectomy status: a randomized, double-blind, placebo-controlled, phase IV study. AB - Flavonoids, such as troxerutin, have been shown to be safe and effective agents for the treatment of chronic venous insufficiency. The fixed combination between troxerutin 150 mg and carbazochrome 1.5 mg (Fleboside ampoules) was previously shown to have a good efficacy and safety profile in non-surgical patients with acute uncomplicated hemorrhoids. The purpose of this randomized, double-blind, placebo-controlled study was to investigate the efficacy and tolerability of the active combination in the treatment of post-hemorrhoidectomy patients. 30 patients were randomized to receive one of two treatments: troxerutin 150 mg and carbazochrome 1.5 mg, or placebo, i.m. 3 ml ampoules twice a day for five consecutive days after the surgical procedure, starting from the day of surgery. Efficacy parameters were assessed as follows: at baseline (T1), after the first administration (T2; day of surgery), the second day after the surgical procedure (T3), and the fifth day after the surgical procedure (T4); hemorrhoidal symptoms based on a visual analogue scale (VAS): pain, discharge, bleeding, inflammation, and pruritus; analgesic intake, if any; time to restore a physiological defecation; edema evaluation (based on a four-point scale: 0 = absent; 1 = mild; 2 = moderate; 3 = severe); camera pictures taken at T1 and T4 (in selected patients); and blood coagulation tests. Analysis between treatment groups revealed a highly significant difference at T3 and T4 for the total VAS score (p = 0.007 and p = 0.001, respectively) in favor of the active combination treatment. A statistically significant difference was also observed for bleeding and pruritus at T3 and for these two parameters and both inflammation and edema at T4 (p < 0.001) in favor of the active combination group. No adverse events were reported. Neither the active combination nor placebo affected blood coagulation tests. We conclude that intramuscular administration of the fixed combination of troxerutin 150 mg and carbazochrome 1.5 mg is effective, well tolerated and superior to placebo in improving hemorrhoidal and post-surgical symptoms during the five days following surgery. PMID- 11922399 TI - Cizolirtine citrate (E-4018) in the treatment of chronic neuropathic pain. AB - This study was performed to determine the efficacy and safety of oral cizolirtine citrate, a novel agent, in the treatment of chronic neuropathic pain. Cizolirtine was tested in a double-blind, placebo-controlled, two-way crossover study, having previously been shown to have significant analgesic and anti-hyperalgesic action in neuropathic pain models and preliminary human studies. Twenty-five patients with neuropathic pain, which was persistent for at least three months, and scored > 30 mm on a 100 mm visual analogue scale (VAS), were included. A subgroup of five patients had primary skin allodynia, i.e. pain evoked by non-noxious stimuli in the territory of the injured nerve. Cizolirtine 200 mg or placebo was administered twice daily for a treatment period of 21 days, each separated by a washout interval of 7 days. Assessments of skin allodynia were performed using the graded monofilaments (von Frey hairs) on days 1 (predose), 14 and 21 (90 min postdose). All patients were instructed to maintain a daily pain diary throughout the study. Results showed that the differences in VAS and allodynia scores between cizolirtine and placebo treatments were not significant in the overall analysis (p > or = 0.05); cizolirtine was well tolerated. In a subgroup of five patients with primary allodynia, a 53% reduction in VAS score from baseline at rest (p = 0.007) and 55% on movement (p = 0.0002) at day 21 was observed with cizolirtine, as compared to 8% at rest (p = 0.5215) and 13% on movement (p = 0.4187) with placebo. Similarly, allodynia improved with cizolirtine (p = 0.03) but not with placebo (p = 0.9) in this subgroup. Cizolirtine may be effective in primary allodynia after peripheral nerve injury, and a further trial in a larger number of such subjects is warranted. PMID- 11922400 TI - Atorvastatin plus pravastatin for the treatment of heterozygous familial hypercholesterolaemia--a pilot study. AB - The most common side-effect of statins, mainly during dose titration, is liver toxicity, In these cases, sufficient control of low density lipoprotein cholesterol (LDL-C) in patients with heterozygous familial hypercholesterolaemia (HFH) becomes problematical. In patients with intolerance to resins as well, especially in the presence of coronary artery disease (CAD), it is practically impossible to reach the LDL-C treatment goal. This study included seven HFH patients with CAD, who presented with alanine amino transferase levels greater than three times the upper normal limit during dose titration of atorvastatin or simvastatin of from 20 mg/day to 40 mg/day. They could not tolerate concomitant cholestyramine administration, and presented with LDL-C levels significantly higher than the treatment goal (100 mg/dl; 2.6 mmol/l). In these patients, a combination of two statins with different pharmacokinetics (20 mg/day of atorvastatin plus 40mg/day of pravastatin) was administered for a mean period of one year. Efficacy was compared with that of monotherapy with each drug alone and with that of 40 mg of atorvastatin in 13 patients, who could also not tolerate resin co-administration, and that of 40 mg/day of atorvastatin plus 12 g of cholestyramine in 30 patients, with similar pretreatment LDL-C levels. No increase in serum transaminases and no symptom or sign of myopathy was recorded during the administration of the combination of the two statins for a mean period of 12 months. The atorvastatin plus pravastatin regimen was more effective than both monotherapies and equally effective with the 40 mg of atorvastatin and the 40 mg of atorvastatin plus 12 g of cholestyramine regimens in reducing LDL-C (59% vs. 57% and 61%, respectively) and triglyceride levels (31% vs. 32% and 28%, respectively), while it also had a better effect on high density lipoprotein cholesterol (13% vs. 7% and 8%). The data suggest that the atorvastatin pravastatin combination has a highly beneficial effect on all lipid parameters, without causing hepatotoxicity, in HFH patients with CAD who are sensitive to higher doses of statins in monotherapy. These results require confirmation in larger studies. PMID- 11922401 TI - The effects of single and repeated administration of ebastine on cognition and psychomotor performance in comparison to triprolidine and placebo in healthy volunteers. AB - OBJECTIVE: The cognitive and psychomotor effects of 10 mg, 20 mg and 30 mg ebastine, a second generation H1-receptor antagonist, were compared with sustained release triprolidine 10 mg (as a verum) and placebo in 10 healthy volunteers in a double-blind, randomised cross-over study. METHODS: Following each dose, subjects were required to perform a series of tests of cognitive function and psychomotor performance at 1 h, 2 h, 4 h and 8 h post-dose on days 1 and 5. The test battery consisted of critical flicker fusion, choice reaction time, simulated car tracking task, Sternberg memory scanning task, assessment of subjective sedation (LARS) and subjective evaluation of sleep (LSEQ). RESULTS: Ebastine at all doses investigated was not statistically significant from placebo in any of the objective tests used. However, as expected for a positive internal control, a number of objective measures were significantly disrupted by triprolidine (p < 0.05). Triprolidine produced an overall increase of the peripheral reaction time component of the simulated car tracking task (SCTT), the difference with placebo reaching statistical significance on day 1, 8 h post-dose (p < 0.05). The mean tracking accuracy scores were also significantly impaired following the administration of triprolidine after 8h on day 1 (p < 0.05). Triprolidine also produced a clear decrement on the SMST (Sternberg Memory Scanning Task), which was significantly different from placebo, at 4 h and 8 h post-dose on day 1. Subjective reports of sedation (LARS) were significantly greater at 2 h and 4 h following triprolidine administration on day 1 and ebastine (30 mg) was rated as sedative 4 h following administration on day 5. The perceived sedative activity of ebastine 30 mg was also reflected in the subjective reports on the LSEQ on day 1 (p < 0.05). CONCLUSIONS: These results allow the conclusion that ebastine, at its recommended therapeutic doses of 10-20 mg, is demonstrably free from impairment on objective aspects of psychomotor and cognitive function in a study where the psychometric assessments were shown to be sensitive to disruptive effects, as evidenced by the action of the positive control, triprolidine 10 mg. PMID- 11922402 TI - Pure antiestrogens and breast cancer. AB - Tamoxifen, which is the most commonly used drug for treatment of breast cancer, has both estrogen agonist and antagonist actions. Pure antiestrogens are devoid of any estrogen agonist effects. ICI 182,780 (fulvestrant) (Faslodex) and ICI 164,384 are competitive inhibitors of estrogen by binding to the estrogen receptor (ER). Preclinical and clinical studies show that fulvestrant and ICI 164,384 are more potent than tamoxifen in inhibiting the growth of breast cancer cells. They are devoid of any estrogen-agonist action on the uterus and vagina but lack the beneficial effects of tamoxifen on the bone and serum lipid profile. Fulvestrant is the first pure antiestrogen to complete phase III clinical trials. Such studies have shown that fulvestrant is at least as good as anastrozole in the treatment of post-menopausal women with advanced breast cancer who had relapsed or progressed on prior endocrine therapy. The drug was well tolerated and only minor side-effects were reported. Its potential role in the adjuvant setting will be determined by its adverse effects on bone mass and serum lipids. EM-800 and EM-652 are the most potent pure antiestrogens and EM-652 has the highest affinity of all antiestrogens to ER. They have no stimulatory effects on the uterus or vagina. It seems reasonable to expect that pure antiestrogens will be good alternatives to tamoxifen and aromatase inhibitors in the treatment of breast cancer. PMID- 11922404 TI - Assessing the impact of migraine. AB - Migraine is a remarkably disabling condition, although unpredictable and heterogeneous in frequency, duration and severity. It can be difficult to manage in primary care, where it is under-recognised, under-diagnosed and under-treated. Proposals have been made that migraine care could be improved by incorporating assessments of migraine impact into management strategies. Research has shown that measuring headache-related disability, together with assessments of pain intensity, headache frequency, tiredness, mood alterations and cognition can be used to assess the impact of migraine on sufferers' lives and society. From this research, two simple and brief impact tools were developed: the Migraine Disability Assessment (MIDAS) Questionnaire and the Headache Impact Test (HIT). Both tools are scientifically valid measures of migraine severity and have the potential to improve communication between patients and their physicians, assess migraine severity and act as outcome measures to monitor treatment efficacy. Each of these tools offers its own advantages. For example, HIT was designed for greater accessibility (on the Internet at www.headachetest.com and www.amlhealthy.com, and as a paper-based form known as HIT-6) and has a wider coverage of the spectrum of headache than MIDAS. Impact tools are also being increasingly recommended as part of generalised headache management guidelines to produce an individualised treatment plan for each patient in concert with other clinical assessments. It is not possible as yet to recommend unequivocally the optimal impact tool for use in primary care, but it should be usable by GPs, pharmacists, nurses and patients, and for research purposes. PMID- 11922403 TI - Comparative haemodynamic responses to the first dose of short- and long-acting ACE inhibitors in patients with congestive heart failure. AB - BACKGROUND: Angiotensin Converting Enzyme inhibitors (ACEi's) confer significant mortality and morbidity benefits in all functional grades of chronic heart failure (CHF). However, physicians' concerns regarding the possible occurrence of first-dose hypotension appear to be a contributing factor to their under utilisation in both hospital and primary care settings. We investigated whether long-acting and short-acting ACEi's differ in their haemodynamic responses to the first-dose in patients with CHF. METHOD: This was a multicentre, randomised, open, two-parallel-group study of captopril 6.25 mg and perindopril 2 mg. 240 patients with CHF, age 68.9 +/- 8.9 years, of whom 66% were male, NYHA II-IV, with average blood pressure baseline values of 132.2 +/- 16.2/78.5 +/- 10.5 mmHg for systolic and diastolic blood pressure, and left ventricular ejection fraction (LVEF) of 31.3 +/- 7.4% received either captopril (n = 124) or perindopril (n = 116). Blood pressure was continuously monitored during the 8 h following drug intake. Minimum levels and maximum decreases in systolic, diastolic and mean arterial pressures were measured, along with the incidence of hypotensive episodes defined as mean blood pressure (MBP) fall > 20 mmHg, whether symptomatic or not. Subgroups of patients distributed according to age, baseline blood pressure (BP) and LVEF were subsequently analysed. RESULTS: Overall, a statistically significant treatment effect in favour of perindopril was observed. First-dose hypotension was observed more frequently following captopril than perindopril administration, with lower MAP minimal levels (78.0 +/- 8.9 vs. 84.5 +/- 10.1 mmHg, p < 0.0001), greater maximum falls (17.6 +/- 8.3 vs. 12.8 +/- 7.3 mmHg, p < 0.0001) and more frequent hypotensive episodes (42% vs. 15%, p < 0.0001). The incidence of at least one symptomatic episode was also significantly higher with captopril (10 patients vs. one patient, p = 0.029). Subgroup analyses according to age (< or = 70 years or > 70 years) or LVEF (< or = 30% or > 30%) reflected the main result. CONCLUSION: Initiation of treatment with ACE inhibitors is associated with different haemodynamic and clinical tolerances in CHF patients, regardless of their risk for hypotension, with possible clinical implications. PMID- 11922405 TI - Does the gloss hide the dross? PMID- 11922406 TI - Urinary fluoride excretion of young children exposed to different fluoride regimes. AB - AIMS: To compare 24-hour urinary fluoride excretion in young children exposed to different fluoride regimes. DESIGN: Twenty-four-hour urine samples were collected from children aged between 1.8 and 5.2 years. Samples were collected from Cork, Ireland (n=19) where the water is fluoridated to a concentration between 0.8 and 1.0 mg/l; Knowsley, UK, where the water fluoride concentration is <0.1 mg/l (n=22); and from children in Knowsley drinking milk containing 0.5 mg fluoride in nursery school each day (n=16). The volume of the samples was measured, they were analysed for fluoride concentration and the 24-hour urinary fluoride excretion was calculated. RESULTS: It was found that the mean fluoride excretion in response to usual conditions of fluoride intake in these children was 0.21 mg (SD=0.14) in non-fluoridated Knowsley; 0.36 mg (SD=0.11) in fluoridated Cork and 0.30 mg (SD=0.10) in the children drinking fluoridated school milk. CONCLUSIONS: The daily fluoride excretion in these children, corrected for age and fluoride ingested from toothpaste, appeared to indicate that the fluoride intake in the children drinking fluoridated school milk was somewhere between those living in an optimally fluoridated area and those in a low fluoride area. PMID- 11922407 TI - Predictors of Norwegian dentists' decisions to recommend replacement of teeth at the time of extraction. AB - OBJECTIVE: To study predictors of dentists' recommendations to replace the teeth which they extracted for adult patients in Norway in 1998. DESIGN: Cross sectional postal questionnaire survey. PARTICIPANTS: A systematic random sample of 1,500 Norwegian general dental practitioners and response rate of 67% after two reminders. After exclusions, 302 dentists who had extracted 822 permanent teeth for 587 patients (16-92 years old) over a period of two weeks, were accepted for the study. RESULTS: According to the dentists, 30% of the patients were in definite need of a replacement for the extracted tooth or teeth, 29% were cases open to question and 41% had no need for a replacement. Model I: Tooth type extracted and the number of teeth extracted per patient were the most important predictors of the dentists definitely recommending a replacement after tooth extraction. Model II: When including the group of patients for whom a replacement was open to question, the odds of the dentists recommending a replacement for having an extracted anterior tooth or premolar increased from 13 (95% CI 6.16, 26.07) to 21 (95% CI 10.51, 40.69). Demographic characteristics had no significant effect on the dentists' decision. CONCLUSIONS: The two most important predictors for recommending a replacement after extraction were the position of the tooth or teeth and the number of teeth extracted. The reason why the number of remaining teeth had no significant independent effect was possibly because this group of patients had a relatively high number of natural teeth. PMID- 11922408 TI - Mothers' sense of coherence and their adolescent children's oral health status and behaviours. AB - OBJECTIVE: To investigate the relationship between mothers' sense of coherence (SOC) and their adolescent children's oral health. SOC is the central construct of the salutogenesis model of health, which seeks to explain factors that promote health. METHOD: A cross-sectional study was conducted in Goiania-GO, Brazil, on 664 15-year-old children randomly selected from schools, and their mothers. Data were collected through questionnaires, the short version of Antonovsky's SOC Scale (13-item) and clinical dental examinations. Multiple logistic regression and polytomous ordered regression were used in the data analysis. Two sets of outcome variables were selected for the analyses: oral health status (dental caries, oral cleanliness, and periodontal disease), and oral health-related behaviours (daily frequency of sugar intake, toothbrushing frequency, and pattern of dental attendance). RESULTS: Adolescents whose mothers had higher levels of SOC score had lower levels of dental caries and gingival bleeding after probing, and were less likely to visit the dentist mainly when in trouble than those whose mothers had lower levels of SOC. CONCLUSION: Mothers' SOC was associated with their children's levels of dental caries and periodontal disease, as well as pattern of dental attendance. Interventions designed to improve or maintain the oral health of young people should take into consideration the family environment. PMID- 11922409 TI - Multilevel survival analysis of amalgam restorations amongst RAF personnel. AB - OBJECTIVE: To introduce the concepts of multilevel survival analysis through an investigation into the longevity of amalgam restorations. BASIC RESEARCH DESIGN: The multilevel Cox proportional hazard model is illustrated using amalgam restoration data comprising three levels: repeated restorations at level-1, teeth at level-2, and subjects at level-3. The outcome was duration of amalgam restoration survival. Single-level and multilevel Cox methods are contrasted. PARTICIPANTS: The data were from a survey of amalgam restorations (reported elsewhere), involving 200 RAF personnel aged between 16 and 37 years at enlistment between 1947 and 1979, having served continuously for a minimum of 16 years prior to 1994. RESULTS: Differences existed between single-level and multilevel methods; the latter being the method of choice. Initial caries experience was a good predictor of longevity. Molar teeth fared worse than pre molars and MOD & B, MOD & L, and MOD & BL restorations experienced considerably greater risk of failure than did MOD, MO, DO and MO/DO ext types, which in turn fared worse than occlusal restorations. Root-treated and pinned teeth also experienced an elevated risk of premature failure. There was a moderate but significant increase in restoration failure amongst subjects who were seen by more dentists throughout their service. CONCLUSIONS: The application of multilevel modelling to survival analysis provides an appropriate and powerful solution to the problem of lack of independence amongst dental restorations. It is beneficial that studies undertake a multilevel analysis in preference to ignoring hierarchy or omitting swathes of information in order to perform a single-level analysis. PMID- 11922410 TI - Self-assessments of general and oral health in persons with chronic whiplash related disorders. AB - OBJECTIVE: The objectives of the study were 1) to describe general health and daily functioning in a group of subjects with whiplash-associated injuries, 2) to analyse self-assessments of oral disease relative to background factors, 3) to investigate if the Medical Outcomes Study Short Form 36 (SF-36) Health Survey variables were related to the oral disease variables. DESIGN: Questionnaires included the SF-36 Health Survey, and self-assessed oral health with regard to relevant background factors such as gender, age, education and marital status. Logistic regression modelling was performed. PARTICIPANTS: Members of a nationwide Swedish association enlisting persons who have problems concerning a whiplash injury (n = 1,928) were included. A total of 979 persons participated in the study, a response rate of 50.8%. RESULTS: More than twice as many females (n=680) as men (n=296) responded to the questionnaire. A general trend, in comparing SF-36 scores between the present study group and the normative sample, was that persons with whiplash associated disorders scored considerably lower on all scales. Older age and being of female gender, together with poorer physical functioning and poorer general health, were related to poorer oral health status. CONCLUSION: Self-assessed oral health was correlated to impairment in general health, a finding which is in agreement with an integrated holistic understanding of human health and disease. One causative chain for this may be through the stress mechanisms. PMID- 11922411 TI - The 80:20 phenomenon: help or hindrance to planning caries prevention programmes? AB - OBJECTIVE: to compare the outcomes of population segmentation analyses according to caries experience using the distribution of dmft and the Super Profiles geodemographic classification. BASIC RESEARCH DESIGN: The study population consisted of all 15,747 children in seven districts in the North West Region, England who were examined in whole population surveys during the 1995/6 NHS epidemiological survey of 5-year-old children. Market penetration analyses were used to segment this population according to caries experience by their dmft score and by the Super Profiles geodemographic classification. Lorenz curves were plotted and Gini coefficients were calculated from the outputs. MAIN OUTCOME MEASURES: When dmft was used to segment the population approximately 80% of carious teeth were found in some 42% of the total population, producing a Lorenz Curve with a Gini coefficient of 66%. The population was then segmented using Super Profiles Target Markets. The target markets were ranked according to caries experience and the denominator population living in each of these area types. Some 81% of the total population dmft was found in the topmost ranked 73.8% of the total population. The resultant Lorenz curve produced a Gini coefficient of 16.2%. CONCLUSIONS: About half of the population disease was confined to a minority of the population but not to the extent of 80% of the disease in 20% of the population. Although these high risk children were more commonly found in underprivileged area types, they did not live exclusively in a small number of deprived areas. These findings shed doubt on the wisdom of a targeted approach to oral health promotion and disease prevention programmes. PMID- 11922412 TI - Quantifying dental inequality--developing the methodology. AB - OBJECTIVE: To develop a simple quantitative measure of dental health inequality. METHOD: Define equality based on statistical theory and construct a formula to compare actual and theoretical disease distributions. RESULT: A new index (DHII) is proposed, based on the ratio of the distribution of the actual disease to a (theoretical) Poisson distribution. An example using caries data is provided. CONCLUSION: Use of DHII will allow quantitative measurement of dental health inequality. PMID- 11922413 TI - The dental caries experience of 12-year-old children in England and Wales. Surveys coordinated by the British Association for the Study of Community Dentistry in 2000/2001. AB - DESIGN: This paper reports the results of standardised clinical caries examinations of 106,694 twelve-year-old children from England, Wales, the Isle of Man and Jersey. These 2000/01 coordinated surveys are the latest in a series which seek to monitor the dental health of children and to assess the delivery of dental services. METHOD: The criteria and conventions of the British Association for the Study of Community Dentistry were used. Representative samples were drawn from participating health authorities and boards, and caries was diagnosed at the caries into dentine (D3) threshold using a visual method without radiography or fibre-optic transillumination. RESULTS: Once again a wide variation in caries prevalence across the United Kingdom was demonstrated, with mean values for D3MFT for the English 'regions' (of the National Health Service) and Wales ranging from 0.63 in the West Midlands to 1.31 in Wales. The mean value for DMFT across England Wales was 0.89 (D3T=0.39, MT=0.07, FT=0.43). Overall, 38% of children had evidence of caries experience at the dentinal level of detection (D3MFT>0), although the means ranged between 30% (West Midlands) and 51% (Wales). The mean D3MFT for those with disease at this threshold was 2.35. Trends over time demonstrate an improvement of 11% in overall D3MFT since 1996/97, compared to the 15% seen over the previous four year period. FT and care index fell. The number of fillings provided in 2000/01 and thus the care index, remained low. On average across England and Wales, only 48% of the dentinal caries experience identified by survey examinations of permanent teeth was seen as fillings (range in individual districts: 30% to 78%). The use of the SiC index highlights dental health inequalities. CONCLUSION: Taken together, these findings demonstrate the continuing need for more effective preventive strategies and treatment services for this important age group. PMID- 11922414 TI - Uptake of trace elements by rice plants inoculated with Pyricularia oryzae. AB - The ability of rice plants inoculated with Pyricularia oryzae (P. oryzae) to take up trace elements was studied by the radioactive multitracer technique. Among various elements, only Mn, Co, Zn, Se, Rb, Sr, Tc, and Re were found to be transferred to rice shoots from soil. The concentrations of essential elements, Mn and Zn, in the shoots of rice plants inoculated with P. oryzae were slightly higher than those in the control plant shoots, while Se, Rb, Tc, and Re showed almost the same concentrations for both the shoots. PMID- 11922415 TI - Surface dose with grids in electron beam radiation therapy. AB - This investigation attempts to solve the problem of the lack of skin-sparing effect in electron radiation therapy and to increase the tolerance of skin to radiation using the grid technique. Electron grid therapy involves the mounting of a Cerrobend grid in the electron cone. Film dosimetry was employed to measure the relative surface dose and the percentage depth dose profile of electron grid portals. Various grid hole diameters (d = 0.45, 1.0, 1.5 cm) and grid hole spacings (s = 0.4, 0.2 cm) were considered for electron beams from 6 to 14 MeV. Experimental results indicate that the electron grid technique can reduce the relative surface dose in electron radiation therapy. Degradations of the relative surface dose depend on the percentage of open area in the grid portal. A proper grid design allows the surface dose to be reduced and the range of nonhomogeneous doses to be limited to a depth at which the target volume can receive a homogeneous dose. The grid technique can lower the surface dose in electron radiation therapy. PMID- 11922416 TI - Optimal design of radioactive particle tracking experiments for flow mapping in opaque multiphase reactors. AB - In the past decade, radioactive particle tracking techniques have emerged in the field of chemical engineering and have become increasingly popular for non invasive flow mapping of the hydrodynamics in multiphase reactors. Based on gamma ray sensitization of an array of scintillation detectors, the Computer Automated Radioactive Particle Tracking (CARPT) technique measures flow fields by monitoring the actual motion path of a single discrete radioactive flow follower which has the physical properties of the phase whose motion is being followed. A limitation to the accuracy of CARPT lies in the error associated with the reconstruction of the tracer particle position which affects the space-resolution capability of the technique. It is of interest, therefore, to minimize this error by choosing wisely the best hardware and an optimal configuration of CARPT detectors' array. Such choices are currently based on experience, without firm scientific basis. In this paper, through theoretical modeling and simulation, we describe how the accuracy of a radioactive particle tracking setup may be assessed a priori. Through an example of a proposed implementation of CARPT on a gas-solids riser, we demonstrate how this knowledge can be used for choosing the hardware required for the experiment. Finally, we show how the optimal arrangement of detectors can be effected for maximum accuracy for a given amount of monetary investment for the experiment. PMID- 11922417 TI - Monte Carlo simulation of surface percent depth dose. AB - This work studied the surface percent depth dose of 6 and 15 MV X-rays, 10 x 10 cm2 and 20 x 20 cm2 fields by Monte Carlo simulation. The OMEGA/BEAM code, an EGS4 user code developed by the NRCC, was used. The linac, Siemens PRIMUS, was accurately modeled according to the ion chamber and CEA film measurement, and the phase space data generated from this linac were collected to simulate dose distribution in water. The water phantom had radius 30 cm and thickness 10 cm. The percent depth doses at zero depth, PDDsurface, for 6 MV X-rays were 13.85 +/- 0.11% and 23.21 +/- 0.20% for the 10 x 10 cm2 and 20 x 20 cm2 fields, respectively. For 15 MV X-rays, PDDsurface values were 8.83 +/- 0.07% and 18.60 +/- 0.12% for the 10 x 10 cm2 and 20 x 20 cm2 fields, respectively. PMID- 11922418 TI - An approach to the evaluation of the activity of the DNA repair enzyme O6 methylguanine-DNA-methyl-transferase in tumor tissue in vivo: syntheses of 6 benzyloxy-9-(2-[18F]fluoroethyl)-9H-purin-2-yl-amine and 6-benzyloxy-7-(2 [18F]fluoroethyl)-7H-purin-2-yl-amine. AB - The resistance of tumor cells to the cytostatic activity of methylating and chloroethylating anticancer drugs is determined by the level of expression of the DNA repair protein O6-methylguanine-DNA-methyl-transferase (MGMT). The synthesis of labelled 6-benzyloxy-9H-purin-2-ylamine derivatives should hence allow a quantification of the MGMT status of tumor and non-target tissue in vivo. 6 benzyloxy-9-(2-fluoroethyl)-9H-purin-2-yl-amine and 6-benzyloxy-7-(2-fluoroethyl) 7H-purin-2-yl-amine were synthesized and evaluated in vitro, both showing an affinity of 1.8 microM. 6-benzyloxy-9-(2-[18F]fluoroethyl)-9H-purin-2-yl-amine and 6-benzyloxy-7-(2-[18F]fluoroethyl)-7H-purin-2-yl-amine were synthesized by alkylation of 6-benzyloxy-9H-purin-2-ylamine with 1-[18F]fluoro-2-tosylethane in optimized yields of 41% and 20%, respectively. Biodistribution studies were performed in nude mice, carrying mex+ (MGMT expressing) and mex- tumors. PMID- 11922419 TI - Electrochemical cell for separation of [18F]fluoride from irradiated 18O-water and subsequent no carrier added nucleophilic fluorination. AB - An electrochemical cell was designed allowing the anodic deposition of n.c.a. [18F]fluoride solubilized in an 18O-water target and subsequent n.c.a. nucleophilic 18F-fluorination. The recovery of the deposited [18F]fluoride can be achieved in the presence of an aprotic solvent containing a phase-transfer catalyst (PTC). The radioisotope adsorbed electrochemically at the cylindrical surface of a glassy carbon electrode can be dried easily by washing the cell twice with dry aprotic solvents while maintaining a low electric field. This simple washing step makes an azeotropic drying process obsolete. Accordingly, less basic cryptates like [K within 2.2.2.]oxalate or triflate can be used for nucleophilic 18F-fluorination because loss of activity as a consequence of azeotropic drying under conditions of low basicity does not occur. The usefulness of this technique is exemplified for the n.c.a. synthesis of various 18F-labelled compounds. The radiotracers were synthesized with higher radiochemical yields and under much easier conditions than the conventional 18F-fluorination procedure which includes an additional drying step of the PTC. PMID- 11922420 TI - Dosimetry of ionizing radiation using an iodide/iodate aqueous solution. AB - Details of a new aqueous chemical dosimeter consisting of a mixture of KI (0.6 M) and the electron scavenger KIO3 (0.1 M) in 0.01 M borate buffer (pH 9.25) are presented. Exposure was either to gamma sources (137Cs, 60Co) or to linear accelerator (LINAC) radiations (18-MeV electrons, bremsstrahlung X-ray spectrum). G values were obtained for the formation of triiodide; the absorbance at its maximum was then measured at 352 nm. The dose response was linear up to 6000 Gy, the lower limit of detection being approximately 0.25 Gy. G values calculated from the initial slopes of the dose-response curves were 14.1 +/- 0.8 for 137Cs radiations and 13.8 +/- 0.4 and 13.9 +/- 0.8 for the NIST and AFRRI 60Co radiations, respectively. G values obtained for the electron and bremsstrahlung radiations were 12.2 +/- 1.9 and 11.9 +/- 1.8, respectively. The iodide/iodate dosimeter extends the range of detection an order of magnitude both above and below the accepted detection limits of the Fricke dosimeter. PMID- 11922421 TI - Improvement of the capabilities of PGAA by coincidence techniques. AB - Applicability of the gamma-gamma regional coincidence method to prompt gamma activation analysis has been examined in a series of model experiments. It is shown that the requirement of coincidence with a spectral range instead of a single peak greatly improves the signal to background ratio, yet preserves the linear relationship between concentration and analytical signal. The method is especially suitable for trace analysis of solutions containing hydrogen, and of matrices containing boron or other strong single gamma-ray emitters. PMID- 11922422 TI - Decay scheme of 126La isomers. AB - The beta+/EC decay of 126La produced by the 94Mo(36Ar, 3pn) reaction has been studied using the on-line isotope separator connected to an AVF cyclotron. Gamma ray singles, gamma-gamma angular correlation and internal conversion electron measurements were carried out. From analysis of these data, 33 new levels in 126Ba were found and the decay scheme containing 137 gamma-rays and 50 excited states was constructed. The probable spin and parity of the two beta-decaying states in 126La were found to be 4+/-, 5+/- for the high-spin isomer with a half life of 54(2) s and 0-, 1+/-, 2- for the low-spin isomer with a half-life shorter than 50 s. PMID- 11922423 TI - Identification and dose assessment of irradiated cumin by EPR spectrometry. AB - The use of electron paramagnetic resonance spectroscopy to accurately distinguish irradiated from unirradiated cumin and assess the absorbed dose to radiation processed cumin is examined. The results were successful for identifying both irradiated and unirradiated cumin. Additive reirradiation of the cumin produces a reproducible dose response function, which can be used to assess the initial dose by back-extrapolation. Third-degree polynomial and exponential functions were used to fit the EPR signal/dose curves. It was found that the 3rd degree polynomial function provides satisfactory results without correction for decay of free radicals. The exponential fit to the data cannot be used without correction of decay of free radicals. The stability of the radiation-induced EPR signal of irradiated cumin was studied over a storage period of 6 months. The additive reirradiation of some samples was carried out at different storage times (10, 20 and 30 days) after initial irradiation. PMID- 11922424 TI - Feeding of the 232Th levels from the decay of 236U. AB - Population of the 6+ 332 keV level in 232Th from the decay of 236U was observed for the first time. The 49, 112, and 171 keV gamma-ray energies and relative intensities from the decay of 236U were measured. PMID- 11922425 TI - The Plasmodium falciparum var gene switching rate, switching mechanism and patterns of parasite recrudescence described by mathematical modelling. AB - Recrudescing Plasmodium falciparum parasitaemia is attributed to the switching of PfEMP1, a variant antigen family encoded by the var gene repertoire, and the host's immune response. We have developed a mathematical model which incorporates var gene switching, and variant specific, non-variant specific and non-specific immunity. By conducting a sensitivity analysis of the model we have defined the parameter limits which produce chronic and recrudescing infections. We explore 3 switching mechanisms: ordered, random and uncoupled switching. We show that if var genes switch on and off independently at variable rates through the repertoire a chronic clinical infection is predicted. The fastest switching-on rate that produces a chronic infection is 0.03% per generation. The model predicts that non-variant specific immunity plays an important role in reducing disease severity. This work illustrates the complex relationship between the malaria parasite and its host and shows that var gene switching at rates substantially slower than 2% are essential for parasite survival. PMID- 11922427 TI - Population dynamics of untreated Plasmodium falciparum malaria within the adult human host during the expansion phase of the infection. AB - A retrospective analysis was performed of parasite count data recorded from the first 7 days of blood or mosquito transmitted Plasmodium falciparum infections given for the treatment of neurosyphilis in the USA before 1963. The objective of this study was to characterize initial growth dynamics before host defences have significant effects on the infecting parasite population. Of the 328 patients' data available for analysis, 83 were excluded because they had received anti malarial treatment during the first 7 days of the patent infection. Nonlinear mixed effects modelling was performed to estimate the parameters of interest; 'parasite multiplication rate per 48 h' (PMR), and length of the parasite life cycle (periodicity). The parasitaemia versus time profiles showed great variability between patients. The mean population estimate of 'PMR' was approximately 8, and was highly dependent on the P. falciparum 'strain'. PMR also varied significantly between patients with a 90% prediction interval varying from 5.5 to 12.3-fold. Both intrinsic parasite multiplication rate (an intrinsic virulence determinant), and host susceptibility and defence contribute to expansion of the parasite biomass and thus disease severity in falciparum malaria. PMID- 11922426 TI - Immunogenic properties of the Plasmodium vivax vaccine candidate MSP1(19) expressed as a secreted non-glycosylated polypeptide from Pichia pastoris. AB - The 19 kDa C-terminal region of the merozoite surface protein 1 (MSP1(19)) is one of the most promising vaccine candidates against the erythrocytic forms of malaria. In the present study, a gene encoding the Plasmodium vivax MSP1(19) epitope (PvMSP1(19)) and the Pan-Allelic DR epitope (PADRE) was expressed in the methylotrophic yeast Pichia pastoris. A non-glycosylated form of the recombinant protein rPvMSP1(19)-PADRE was purified from culture supernatants. This recombinant protein maintains its antigenicity, being recognized by a very high percentage (85.6%) of sera from Brazilian individuals naturally exposed to P. vivax. The antibody immune response elicited by rPvMSP1(19)-PADRE was compared in C57BL/6 mice immunized with different adjuvant formulations. After 3 immunizing doses, antibody titres induced in the presence of the adjuvants monophosphoryl lipid A, trehalose dicorynomycolate and cell wall skeleton or alum plus CpG ODN 1826 were as high as titres generated by Complete Freund's Adjuvant. Based on these immunological studies, we concluded that rPvMSP1(19)-PADRE deserves further evaluation in pre-clinical immunizations against P. vivax in non-human primates. PMID- 11922428 TI - The persistence of Theileria parva infection in cattle immunized using two stocks which differ in their ability to induce a carrier state: analysis using a novel blood spot PCR assay. AB - An improved Theileria parva DNA detection assay based on the polymerase chain reaction (PCR) using primers derived from the 104 kDa antigen (p104) gene was developed to detect parasite DNA in blood spots on filter paper. The specificity of the assay was validated using DNA from a wide range of cattle-derived and buffalo-derived stocks of T. parva. DNA of T. annulata, T. buffeli, T. lestoquardi, T. mutans and T. taurotragi was not amplified using the p104 primers. The detection threshold of the assay was approximately 1-2 parasites/microl of infected blood. PCR amplification using the p104 primers was applied to sequential samples from groups of cattle experimentally infected with either the T. parva Marikebuni stock that induces a long-term carrier state or the Muguga stock, which does not induce a carrier state. The study extended for up to 487 days post-infection and PCR data from defined time points were compared with parasitological microscopy and serological data, together with xenodiagnosis by experimental application of ticks. Microscopy first detected piroplasms between days 13 and 16 after infection whereas all cattle became PCR +ve between days 9 and 13. Animals infected with the Muguga stock of T. parva had parasite DNA in the peripheral blood, which could be detected by PCR, for between 33 and 129 days post-infection in different animals. By contrast parasite DNA in the blood of cattle infected with the Marikebuni stock could be detected consistently from day 9 up to 487 days, when the study terminated. The data suggest that the nature and persistence of the carrier state may differ markedly between different T. parva parasite stocks. PMID- 11922429 TI - Chromosomal size variation in Trypanosoma cruzi is mainly progressive and is evolutionarily informative. AB - The evolutionary significance of chromosome size polymorphism was explored in a representative panel of 26 Trypanosoma cruzi stocks. We tested a progressive model (aCSDI) assuming that the larger the size difference between homologous chromosomes, the more divergent the parasites are. This was contrasted with a non progressive model (Jaccard's distance), in which any chromosome size difference has the same weight. ACSDI-based dendrograms were very similar to those built-up from multilocus enzyme electrophoresis (MLEE) and random amplified polymorphic DNA (RAPD) data: structuring in 2 major lineages (T. cruzi I and T. cruz II) and 5 small subdivisions within T. cruzi II was identical, and branching was very similar. Furthermore, a significant correlation (P < 0.001) was observed between aCSDI and phenetic distances calculated from MLEE and RAPD data. In contrast, analysis of chromosome size polymorphism with Jaccard's distance generated dendrograms with relatively long branches, causing most branching points to cluster close together, which generates statistically uncertain branching points. Our results thus support a model of progressive chromosome size-variation and show that despite an extensive polymorphism, chromosomal sizes constitute valuable characters for evolutionary analyses. Furthermore, our data are consistent with the clonal evolution model previously proposed for T. cruzi. PMID- 11922430 TI - Purification of a 64 kDa antigen from Trypanosoma evansi that exhibits cross reactivity with Trypanosoma vivax. AB - Trypanosoma evansi and Trypanosoma vivax are the most extensively distributed trypanosomes responsible for diseases in livestock. Western blot and indirect immunofluorescence assays revealed a high immunological cross-reaction between these two parasites. An antigen with an apparent molecular mass of 64 kDa (p64), which exhibited cross-reactivity with T. vivax, was purified to homogeneity from a Venezuelan isolate of T. evansi. This antigen is glycosylated, contains a glycosyl-phosphatidylinositol anchor and appeared to be localized through the cell except in the nucleus, indicating that it could primarily be confined to the parasite surface. These results, together with its relative abundance and apparent molecular weight, suggest that p64 probably corresponds to the soluble form of a variable surface glycoprotein from T. evansi. Anti-p64 polyclonal antibodies, raised on mice, recognized a 53 kDa polypeptide band from a Venezuelan isolate of T. vivax on Western blots. Additionally, sera obtained from naturally infected animals also recognized p64, suggesting its potential use as a diagnostic reagent. Mild acid treatment only slightly decreased the immunorecognition of p64, suggesting its potential use as a diagnostic reagent. Mild acid treatment only slightly decreased the immunorecognition of p64, demonstrating that another relevant cross-reacting epitope, different than the inositol-1,2-cyclic phosphate of the cross-reacting determinant, must exist in p64. To date, p64 represents the first antigen isolated and partially characterized from T. evansi. PMID- 11922431 TI - Antibodies induced in mice by a DNA-construct coding for the elastase of Schistosoma mansoni recognize the enzyme in secretions and preacetabular glands of cercariae. AB - A DNA-construct coding for the elastase of the parasite Schistosoma mansoni was prepared from adult S. mansoni worm RNA which was reverse transcribed into cDNA. The gene coding for the elastase was amplified using primers specific for the sequence of cercarial elastase and was cloned into a mammalian expression vector. Expression of the elastase gene at the transcriptional level was achieved for the first time in transfected mammalian cells (COS-7) and was also successful in muscle tissue of mice injected with the DNA-construct. These mice developed antibodies recognizing in Western blots the elastase from cercarial secretions. Also, these antibodies reacted in immunofluorescence tests with the preacetabular glands of cercariae, i.e. the site of origin for elastase. Thus, the DNA construct induced the expression of elastase in mice and formation of antibodies that recognized the native antigen. PMID- 11922432 TI - Comparison of european Trichobilharzia species based on ITS1 and ITS2 sequences. AB - Schistosomes are parasites of considerable medical and veterinary importance and, therefore, all aspects of their biology have been intensively studied. In contrast, our knowledge of species of the largest genus, Trichobilharzia, is insufficient. Because morphological characterization of Trichobilharzia species provides a limited number of criteria for species determination, molecular data are required. In the present paper, we sequenced internal transcribed spacers ITS1 and ITS2, and 5.8S ribosomal RNA (rRNA) genes of 3 European Trichobilharzia species (T. regenti, T. szidati and T. franki). We showed that ITS1 and ITS2 sequences can be used in species identification. Repetitive elements were found in ITS1 of all 3 Trichobilharzia species; their number and length varied depending on the species. Phylogenetic analysis showed that the visceral T. franki is more related to the nasal T. regenti, than to the visceral T. szidati. The newly designed primer, which is specific for T. regenti, might be used as a tool for diagnosis of this potential pathogen. PMID- 11922433 TI - Needle in a haystack: involvement of the copepod PARACARTIA grani in the life cycle of the oyster pathogen Marteilia refringens. AB - Marteilia refringens is a major pathogen of the European flat oyster, Ostrea edulis Linnaeus. Since its description, the life-cycle of this protozoan parasite has eluded discovery. Attempts to infect oysters experimentally have been unsuccessful and led to the hypothesis of a complex life-cycle involving several hosts. Knowledge of this life-cycle is of central importance in order to manage oyster disease. However, the exploration of M. refringens life-cycle has been previously limited by the detection tools available and the tremendous number of species to be screened in enzootic areas. In this study, these two restrictions were circumvented by the use of both molecular detection tools and a mesocosm with low biodiversity. Screening of the entire fauna of the pond for M. refringens DNA was systematically undertaken using PCR. Here, we show that the copepod Paracartia (Acartia) grani is a host of M. refringens. Not only was DNA of M. refringens consistently detected in P. grani but also the presence of the parasite in the ovarian tissues was demonstrated using in situ hybridization. Finally, successful experimental transmissions provided evidence that P. grani can be infected from infected flat oysters. PMID- 11922434 TI - Triclabendazole-resistant Fasciola hepatica: beta-tubulin and response to in vitro treatment with triclabendazole. AB - Resistance in Fasciola hepatica to triclabendazole ('Fasinex') has emerged in several countries. Benzimidazole resistance in parasitic nematodes has been linked to a single amino acid substitution (phenylalanine to tyrosine) at position 200 on the beta-tubulin molecule. Sequencing of beta-tubulin cDNAs from triclabendazole-susceptible and triclabendazole-resistant flukes revealed no amino acid differences between their respective primary amino acid sequences. In order to investigate the mechanism of triclabendazole resistance, triclabendazole susceptible and triclabendazole-resistant flukes were incubated in vitro with triclabendazole sulphoxide (50 microg/ml). Scanning and transmission electron microscopy revealed extensive damage to the tegument of triclabendazole susceptible F. hepatica, whereas triclabendazole-resistant flukes showed only localized and relatively minor disruption of the tegument covering the spines. Immunocytochemical studies, using an anti-tubulin antibody, showed that tubulin organization was disrupted in the tegument of triclabendazole-susceptible flukes. No such disruption was evident in triclabendazole-resistant F. hepatica. The significance of these findings is discussed with regard to the mechanism of triclabendazole resistance in F. hepatica. PMID- 11922435 TI - The osmotic relationship of the swimbladder nematode Anguillicola crassus with seawater eels. AB - The study was designed to test the hypothesis that the majority of individuals in a population of Anguillicola crassus can tolerate seawater by osmoconformation with the blood plasma of the eel host. The osmolality of the pseudocoelomic fluid of pre-adult and adult A. crassus was compared to the osmolality of eel plasma after eels were maintained in laboratory freshwater for 2 weeks (short-term transfer) or 3 months (long-term transfer) or in natural seawater for 2 h (acute transfer), 2 weeks or 3 months. The majority of A. crassus (at least 90% of the tested population) osmoconform with their hosts in seawater within +/-30 mOsm/kg of host osmolality. Some pre-adults and adults (15-21% of the total population) were unable to withstand osmotic stress resulting in vacuolation of the hypodermis and intestinal wall, and cuticular detachment. The reasons for variation in the tissue tolerance of A. crassus to increased osmolality of host plasma are unknown and are not related to maturity, sex or conditions in the swimbladder. Osmoconformation in the majority of the population, however, enables parasites to survive and reproduce for long periods in seawater eels. The adaptation of A. crassus to its euryhaline host has played an important part in the rapid spread of the swimbladder nematode in populations of the European eel. PMID- 11922436 TI - Spectrum and outcome of atrioventricular septal defect in fetal life. AB - OBJECTIVES OF THE STUDY: to analyse the features and outcomes of different types of atrioventricular septal defects, detected during fetal life, as compared to postnatal data. MATERIAL AND METHODS: We analysed retrospectively the data concerning 82 fetuses with atrioventricular septal defect, diagnosed from 19 through 37 weeks gestation with a median of 26 weeks. In 46 cases (56.1%), the diagnosis has been made before the age of 24 weeks. RESULTS: Characteristics of the series--in 44 fetuses the atrioventricular septal defect was not associated with other cardiac anomalies, while 38 fetuses had a more complex form. Chromosomal anomalies were present in 33 of the fetuses (40.2%), more frequently in cases without associated intracardiac defects (56.8%). Trisomy 21 occurred in just over one quarter the series, and in 43.2% of cases without associated defects. In addition, 11% of fetuses had trisomy 18, and one had trisomy 13. Extracardiac anomalies were present in 12 of the fetuses (14.6%), more frequently in cases without associated abnormalities. Of fetuses with more complex defects, 46.4% had hypoplasia of the left ventricle and aorta. Complete atrioventricular block was present in 10 of the fetuses (12.2%), mainly in fetuses with other malformations, and particularly with left isomerism. Recurrence of congenital heart disease was observed in 5 of the fetuses (6.1%). OUTCOME: In 25 instances (30.5%) the parents opted for termination of pregnancy. Of 57 cases that continued through pregnancy, 9 fetuses died prior to term (15.8%), 32 died postnatally (56.13%) and only 16 fetuses (28.1%) survived. Overall, the mortality was higher in cases with associated malformations, in those with heart failure or those with atrioventricular block. Cardiac surgery was performed in 19 infants, with 5 dying postoperatively, and one late. CONCLUSIONS: Our data show a high prevalence of atrioventricular septal defect associated with other malformations when diagnosed during fetal life. This combination is less frequently associated with chromosomal and extracardiac anomalies, but more often with obstructive lesions of the left heart and with atrioventricular block. The association results in a less favourable outcome. PMID- 11922437 TI - Cleft of the mitral valve in patients with Down's syndrome. AB - Differentiation between a cleft of the mitral valve and the cleft of the left side of an atrioventricular septal defect--a lesion commonly found in patients with Down's syndrome--is surgically important since the distribution of the conduction tissue varies between the 2 lesions. We sought to determine if cleft of the mitral valve occurs also in patients with Down's syndrome. We studied 5 patients with Down's syndrome and cleft of the mitral valve followed in our institution. Echocardiography showed in all 5 patients a cleft dividing the anterior (aortic) leaflet of mitral valve with normal papillary muscle position, mural leaflet size, and ratio of the inlet/outlet dimension of the left ventricle. Associated cardiac lesions were present in all 5 patients: perimembranous ventricular septal defect in 3, ostium secundum atrial septal defect in 2 and patent ductus arteriosus in 2 patients. During the 5.6 years (0.2 11) of the follow-up period, surgical repair of the cleft was never indicated since the mitral regurgitation through the cleft remained mild or absent in all the patients. Two patients underwent closure of a ventricular septal defect, with atrial septal defect closure in one and ductal ligation in 2. One patient died suddenly at home, without evidence of a cardiac cause. In conclusion, a cleft of the mitral valve has important developmental and morphologic differences with atrioventricular septal defect and may occur in patients with Down's syndrome. If surgical repair of the cleft or of associated cardiac lesion is indicated, it is necessary to distinguish it from atrioventricular septal defect where the conduction axis is displaced posteriorly and may be exposed during surgery. PMID- 11922438 TI - In pursuit of excellence: monitoring results and assessment of performance in congenital cardiac surgery. PMID- 11922439 TI - The T wave as a marker of dispersion of ventricular repolarization in premature infants before and while on treatment with the I(Kr) channel blocker cisapride. AB - AIMS: Measurement of electrocardiographic intervals to assess dispersion in ventricular repolarization may be helpful in the assessment of the risk of ventricular arrhythmia. We measured QTc, QT dispersion, and T wave intervals in premature infants before and while on treatment with the I(Kr) blocker cisapride as markers for dispersion in ventricular repolarization. METHODS AND RESULTS: We enrolled 15 non-ventilated premature infants with a mean gestational age of 30.5 weeks, ranging from 26.5 to 33.5 weeks, and mean postnatal age of 24 days, with a range from 5 to 51 days. A digital 12 lead electrocardiogram was recorded prior to and 3 days after administering cisapride at a dose of 0.8 mg/kg/day. Serum electrolytes were simultaneously measured. Electrocardiographic measurements before and after included: QT, QTc Bazett, QT dispersion, R-R, T wave interval peak to end, T wave interval peak to end/onset Q to T wave peak, T wave axis, T wave maximum voltage and QRS-T angle. A paired t test and analysis of variance was used to compare the variables before and during treatment. The QTc, T wave interval peak to end and the ratio T wave interval peak to end/onset Q to T peak increased significantly following treatment with cisapride. Results expressed as before and during treatment were for QTc: 429 (65) ms versus 454 (29) ms p < 0.02; for T wave interval peak to end: 65 (11) ms versus 103 (24) p <0.01, for the ratio T wave interval peak to end/onset Q to T peak: 0.32 (0.06) versus 0.55 (0.16) p < 0.001. Treatment with the I(Kr) blocker did not significantly alter the QT dispersion, T wave voltage, angle or QRS-T angle. CONCLUSION: The interval from the peak to the end of the T wave and the ratio of this value to the onset Q to T peak interval, represents regional dispersion of repolarization across the ventricular wall. This is a potentially useful clinical index in the assessment of arrhythmic risk in premature infants being treated by blockade of the I(Kr) channels. PMID- 11922440 TI - Identification of gender differences in the thickness of the left ventricular wall by echocardiography in children. AB - We assessed the thickness of the normal left ventricular wall according to gender by performing two-dimensional echocardiography in children. While a few studies have evaluated left ventricular wall thicknesses in children using two dimensional echocardiography, there is little information related to the influence of gender throughout childhood. We evaluated 251 patients (128 males and 123 females) with innocent murmurs but without congenital heart anomalies, or coronary artery lesions or arrhythmias, as determined by repeated echocardiographic, and electrocardiographic examinations. Intra-ventricular septal thicknesses at end-diastole and end-systole, and left ventricular posterior thicknesses at end-diastole and end-systole were determined in the parasternal short-axis view. Significant gender differences were observed in the thickness of all four measurements. There was a linear relationship between the thickness of the left ventricular wall and height. Height was the most appropriate and practical index for assessing the thickness of the wall of the left ventricle in children. PMID- 11922441 TI - Three-dimensional computed tomography in children with compression of the central airways complicating congenital heart disease. AB - PURPOSE: We investigated the quality and usefulness of spiral computed tomography and three-dimensional reconstruction in children with obstruction of the central airways as a complication of congenital heart disease. MATERIALS AND METHODS: Spiral computed tomography with three-dimensional reconstruction was performed in 49 children with obstruction of the central airways seen as a complication of congenital heart disease. Spiral scanning was performed during breathing in 40, and after sedation with chloral hydrate in 38. Contrast medium was administered through a pedal venous route in 42. We analyzed the motion artifact, additional information provided by, and clinical usefulness, of the three-dimensional images. We also investigated the factors influencing the quality of the images. RESULTS: Stenoses were seen in the trachea in 21 patients, and in bronchuses in 28. Their causes were an anomaly of the aortic arch in 6, posterior displacement of the aortic arch in 7, posterior displacement of the ascending aorta in 5, compression of the brachiocephalic artery in 5, absent pulmonary valve syndrome in 6, displaced or dilated cardiovascular structure in 17, and pulmonary arterial sling in 2. Motion artifact caused mild or negligible degradation of images in all patients except 6. Breath-holding in non-sedated children produced more severe motion artifact than did cardiovascular pulsation. Three-dimensional images provided additional information over two-dimensional images in 11, and provided clinically useful information in 10. Contrast injection via the pedal route was better for the quality of three-dimensional images than brachiocephalic injection (p = 0.013). CONCLUSIONS: Three-dimensional computed tomography is useful in evaluation of obstruction of the central airways in children with congenital heart disease. Despite the fact that motion artifact is unavoidable, the quality of three-dimensional images is acceptable for making a proper and accurate diagnosis. A pedal route is recommended for injection of contrast medium. PMID- 11922442 TI - Coil occlusion of the small patent arterial duct without arterial access. AB - BACKGROUND: Arterial access is traditionally considered mandatory during coil occlusion of the patent arterial duct. Arterial access necessitates heparinization and carries the risk of femoral artery occlusion in small children. METHODS AND RESULTS: Between July 1999 and May 2001, we attempted coil occlusion of patent arterial ducts in 104 patients without arterial access. The patients were aged from 3 months to 14 years. The median age was 2 years. They weighed 3-35 kg. The median weight was 9.8 kg. The duct diameter at pulmonary artery insertion was 1.8-3.5 mm. The patients were selected on basis of echocardiographic evaluation of duct diameter at pulmonary artery insertion and morphology of the ampulla. Doppler color flow imaging was used in the catheterization laboratory to confirm duct closure. Arterial access was required in 21 patients. The reasons included accidental puncture in 5 patients, failure to obtain venous access in 1 patient, aortic embolization in 3 patients, poor echo images in 2 patients, requirement for additional coils in 8 patients and, failure to cross the duct from pulmonary artery in 2 patients. The fluoroscopic time ranged from 2.2 to 20 min with a mean of 5.3+/-3.8 min. Immediate closure was achieved in 98 patients and this included 79 of the 83 patients in whom arterial access was avoided. Color Doppler 3-24 h later showed residual flow in 2 patients. Four patients had new-onset left pulmonary artery turbulence with peak gradients below 5 mm of mercury. Coil embolization occurred in 6 patients and all coils were retrieved. Three-month follow up information was available for 78 patients. Small residual ductal leaks were seen in 4 patients, 2 of whom had leaks at 24 h. Two patients had recanalized their ducts. CONCLUSION: It is feasible to occlude small patent arterial ducts with coils using venous access alone in carefully selected patients with excellent immediate and short-term results. PMID- 11922443 TI - Congenital atresia of the orifice of the left coronary artery. AB - A 3-month-old infant developed signs of cardiac failure, which was initially attributed to cardiomyopathy. At 8 months, further investigations showed evidence of myocardial ischaemia with reversal of the flow of blood in the left coronary artery, which received no demonstrable inflow from the aorta. An anomalous connection of this artery with the pulmonary trunk was diagnosed but, at surgery, it was found that the arterial orifice was completely atretic, although the main stem was of normal size. A left internal thoracic arterial graft to the anterior descending coronary artery was performed, but he died on the third day after the operation. Postmortem examination showed a small dimple within the aorta at the site of the orificial atresia, extensive myocardial infarction, and two zones of myocardial bridging of the anterior descending coronary artery. We discuss the relationship of coronary orificial atresia with single coronary artery. Although they are related, they typically have different and contrasting clinical presentations. The possible role of the myocardial bridging is also considered. PMID- 11922444 TI - Images in congenital heart diseases. Documentation of decreased adrenergic receptors in left heart failure complicating aortic coarctation in infancy. PMID- 11922445 TI - Images in congenital heart disease. Partial pentalogy of Cantrell. PMID- 11922446 TI - Multiple valvar replacements for hypereosinophilic syndrome. AB - A boy with familial eosinophilia had the hypereosinophilic syndrome, with involvement of mitral and tricuspid valves. Between the ages of 11 and 20 years, he underwent eight surgical procedures on his atrioventricular valves. The pathology included recurrent thrombotic vegetative masses related to hypereosinophilia. Initial repair of the mitral valve was shortlived, but recurrent repairs of the tricuspid valve were helpful. Mechanical prostheses inserted in the mitral position thrombosed despite anticoagulant therapy, and bioprosthetic valves deteriorated with thrombus, fibrosis, or tearing. The hypereosinophilic syndrome is unusual in children, and produces additional problems with valvar surgery. PMID- 11922447 TI - Aortopulmonary collateral arteries in a child with trisomy 21. AB - We describe an infant born prematurely at 30 weeks gestation with Down's syndrome who became dependent on oxygen at 3 weeks of age after an uneventful initial neonatal period. There had been no evidence of bronchopulmonary dysplasia. An isolated aortopulmonary collateral artery of moderate size was mistakenly diagnosed as persistent patency of the arterial duct on echocardiography, and subsequently successfully occluded using two coils with an excellent clinical result. To the best of our knowledge, this is the first description of a congenital aortopulmonary collateral artery in a symptomatic infant with Down's syndrome and no evidence of bronchopulmonary dysplasia. We discuss the possible etiologies of these collateral arteries. It is important to include aortopulmonary collateral arteries in the differential echocardiographic diagnosis of an arterial duct. PMID- 11922448 TI - Exertional pulmonary edema revealing anomalous origin of the left coronary artery from the right coronary aortic sinus. AB - We report a 12-year-old child with anomalous origin of the left coronary artery from the right coronary aortic sinus, the artery taking a proximal intramural course. The anomalous artery was reimplanted into the left coronary aortic sinus. Postoperative stenosis was successfully treated with percutaneous transluminal angioplasty and implantation of a stent. PMID- 11922449 TI - Use of an Amplatzer ductal occluder to close a persistent left superior caval vein which reopened after a total cavopulmonary anastomosis. AB - A 7-year-old boy developed increasing cyanosis after a total cavopulmonary connection with a 3 mm fenestration in the baffle. Catheterisation performed 4 years and 7 months after the operation showed reopening of a left superior caval vein draining into the pulmonary venous atrium. Due to the large size of the left superior caval vein, and the absence of intrinsic stenosis, we chose to use an Amplatzer ductal device to occlude the reopened vein. The procedure was safe and successful. PMID- 11922450 TI - Re: Reduction in levels of triidothyronine following the first stage of the Norwood reconstruction for hypoplastic left heart syndrome. PMID- 11922451 TI - Re: Successful treatment using heparin of the bronchial cast syndrome after the Fontan procedure. PMID- 11922452 TI - Re: Diuretics in the treatment of bronchial casts in congenital heart disease. PMID- 11922455 TI - Sequential dependence of freeze-drying and irradiation on biomechanical properties of rat bone. AB - Femurs of Sprague-Dawley rats were subjected to in vitro 3-Mrad irradiation and/or freeze-drying to investigate whether these processes have an order dependent effect on the biomechanical properties of bone. Four experimental groups were designated-irradiated (RAD); freeze-dried (FD); irradiated, then freeze-dried (RAD/FD); and freeze-dried, then irradiated (FD/RAD). After the various treatments, the bones were inspected for microfractures and then torsion tested. Microfractures were seen in more than 90% of the specimens that had undergone freeze-drying alone or with irradiation. Regarding the normalized relative ratios of torque, there was a statistically significant difference (P < .05) between the RAD group (1.0) and the 3 other groups (FD, .32; RAD/FD, .40; FD/RAD, .14). Differences among the FD, RAD/FD, and FD/RAD groups were not significant. However, a trend was bones were weaker than FD and noted: FD/RAD RAD/FD bones. PMID- 11922454 TI - Vertebroplasty: a new treatment strategy for osteoporotic compression fractures. AB - Vertebroplasty is a procedure in which bone cement is injected into a fractured vertebral body in an attempt to stabilize fractured segments and reduce pain. This procedure was originally used to treat spinal lesions caused by metastases and has recently been used to treat severe bone loss caused by osteoporosis. In this article, we review the current treatment for osteoporosis, introduce vertebroplasty with its associated efficacy and risks, and describe kyphoplasty. PMID- 11922456 TI - Rhodotorula minuta: an unusual fungal infection in hip-joint prosthesis. AB - Fungal infections of total joint arthroplasty are uncommon complications, and most cases have been associated with Candida species. In this article, we present a case in which a total hip arthroplasty was infected with Rhodotorula minuta. Intraoperative gram stain showed yeast formsand led to early detection and immediate treatment with amphotericin B and bacitracin. Successful reimplantation was performed 12 months later. To our knowledge, this is the first reported case of arthroplasty infection caused by R minuta. PMID- 11922457 TI - Acute calcific discitis in adults. AB - Acute calcific discitis in the pediatric population is a well-described entity and may rarely involve the adult population. Acute calcific discitis does occur in adults, presenting clinically with the abrupt onset of severe back pain with a classic radiographic calcification of the disc similar to that of a discogram; it resolves with nonsurgical symptomatic treatment. To our knowledge, no cases of "idiopathic" adult calcific discitis have been reported in the orthopedic literature. We report 2 adult patients with symptomatic thoracic disc calcifications that closely resemble those observed in children and briefly discuss the relevant clinical and radiographic features, along with a review of the literature. PMID- 11922458 TI - Thoracolumbar flexion-distraction injuries combined with vertebral body fractures. AB - Surgery was performed on 25 patients with combination injuries (flexion distraction injury plus vertebral body fracture): 8 patients with anterior-column failure (compression) and 17 patients with anterior-column plus middle-column failure (burst). Patients with compression received posterior instrumentation and underwent fusion; patients with burst received posterior instrumentation and later underwent anterior decompression and fusion. Eleven patients in the burst group had a neurologic deficit. Single dural tears were discovered in 7 patients during the posterior-instrumentation procedure. By the end of the follow-up period (mean, 34.4 months; range, 18-76 months), neither implant failure nor loss of correction had occurred. Combined mechanisms may go unrecognized and thereby result in increased morbidity and inappropriate treatment. Proper evaluation of the posterior elements is of utmost importance for the diagnosis of flexion distraction injuries with vertebral body fractures. After diagnosis, treatment should be started with a posterior procedure. PMID- 11922459 TI - Unconventional alternative for hook plate fixation of the cervicothoracic junction. AB - The most common cause for instability at the cervicothoracic junction (C7-T1) is traumatic fracture-dislocation. Surgical treatment of these unstable injuries has gained wide acceptance as the treatment of choice. The hook plate is the device of choice for achieving dorsal fusion of these injuries. It guarantees independent stability that is sufficient even in cases without anterior stabilization. Because of the high cost of commercially available hook plate systems, we resorted to the use of modified semi-tubular plates for posterior fixation of the cervicothoracic junction. Semi-tubular plates are widely available, cost effective, and can be easily modified to function as a hook plate. PMID- 11922460 TI - Knee arthoplasty: recently developed ceramics offer new solutions. AB - Knee arthroplasty has reached a high degree of reliability. Many of the remaining long-term loosening cases are due to the polyethylene wear debris. The introduction of the alumina ceramic to polyethylene bearing couple had been a promising means for its reduction. The discussion of its applications in total knee replacements shows the possibilities and limitations of this bearing couple. The complete avoidance of the polyethylene debris can be expected from the articulation of alumina ceramic against itself, as long-term follow-up studies of total hip replacements clearly indicate. However, this combination can only be considered with a design of the prostheses in which any incongruity of the articulating surfaces can be excluded. As this necessitates the introduction of an additional component into the relatively narrow space between the femoral condyle and the tibia replacement, a further condition is the availability of a material which, whilst maintaining the tribological advantages of the alumina ceramic, has a much higher mechanical strength than the medical grade alumina ceramic available before. As the specifications of a recently developed modification of an alumina ceramic are in the desired range, with an improvement by a factor of two for nearly all relevant properties, a total knee replacement in which all articulating surfaces are in complete congruity and consist of this extremely wear resistant modified alumina ceramic could be designed, manufactured, and tentatively implanted in a human cadaver knee. PMID- 11922462 TI - Improvement in the morphology of Ti-based surfaces: a new process to increase in vitro human osteoblast response. AB - Surface roughness has been shown to be an influencing parameter for cell response. In this experience we attempted to compare the effect of roughness organization of Ti6A14V or pure titanium substrates on human osteoblast (hOB) response (proliferation, adhesion). Surface roughness was extensively analyzed at scales above the cell size (macro-roughness) or below the cell size (micro roughness) by calculation of relevant classic amplitude parameters (Ra, Rt) and original frequency parameters (Order, Delta). We developed a new process to prepare isotropic surfaces (electro-erosion), which were compared to isotropic surfaces obtained by polishing and anisotropic surfaces obtained by machine tooling. The hOB response on electro-eroded (EE) Ti6A14V surfaces or pure titanium (Ti) surfaces was largely increased when compared to polished or machine tooled surfaces after 21 days of culture. Moreover, the polygonal morphology of hOB on these EE surfaces was very close to the aspects of hOB in vivo on human bone trabeculae. By a complete description of the surface topography of EE surfaces, we concluded that when the topography was considered below the cell scale, hOB appreciated their isotropic smooth aspect, although when the topography was considered above the cell scale they appreciated their rough isotropic 'landscape' formed by many 'bowl-like nests' favouring cell adhesion and growth. Electro-erosion is a promising method for preparation of bone implant surfaces, as it could easily be applied to preparation of most biomaterials with complex geometries. PMID- 11922461 TI - Surface modification of superparamagnetic magnetite nanoparticles and their intracellular uptake. AB - Superparamagnetic magnetite nanoparticles were surface-modified with poly (ethylene glycol) (PEG) and folic acid, respectively, to improve their intracellular uptake and ability to target specific cells. PEG and folic acid were successfully immobilized on the surfaces of magnetite nanoparticles and characterized using fourier transform infrared spectra. The nanoparticle internalization into mouse macrophage (RAW 264.7) and human breast cancer (BT20) cells was visualized using both fluorescence and confocal microscopy, and quantified by inductively coupled plasma emission spectroscopy (ICP). After the cells were cultured for 48 h in the medium containing the nanoparticles modified with PEG or folic acid, the results of fluorescence and confocal microscopy showed that the nanoparticles were internalized into the cells. The ICP measurements indicated that the uptake amount of PEG-modified nanoparticles into macrophage cells was much lower than that of unmodified nanoparticles. while folic acid modification did not change the amount of the uptake. However, for breast cancer cells, both PEG and folic acid modification facilitated the nanoparticle internalization into the cells. Therefore, PEG and folic acid modification of magnetite nanoparticles could be used to resist the protein adsorption and thus avoid the particle recognition by macrophage cells, and to facilitate the nanoparticle uptake to specific cancer cells for cancer therapy and diagnosis. PMID- 11922463 TI - Effect of filler content on mechanical and dynamic mechanical properties of particulate biphasic calcium phosphate--polylactide composites. AB - A bioabsorbable self-reinforced polylactide/biphasic calcium phosphate (BCP) composite is being developed for fracture fixation plates. One manufacturing route is to produce preimpregnated sheets by pulling polylactide (PLA) fibres through a suspension of BCP filler in a PLA solution and compression moulding the prepreg to the desired shape. To aid understanding of the process, interactions between the matrix and filler were investigated. Composite films containing 0 0.25 volume fraction filler, produced by solvent casting, were analysed using SEM, tensile testing and dynamic mechanical analysis (DMA). Homogeneous films could be made, although some particle agglomeration was seen at higher filler volume fractions. As the filler content increased, the failure strain decreased due to a reduction in the amount of ductile polymer present and the ultimate tensile strength (UTS) decreased because of agglomeration and void formation at higher filler content. The matrix glass transition temperature increased due to polymer chain adsorption and immobilization onto the BCP particles. Complex damping mechanisms, such as particle-particle agglomeration, may exist at the higher BCP volume fractions. PMID- 11922464 TI - Ocular disposition and tolerance of ganciclovir-loaded albumin nanoparticles after intravitreal injection in rats. AB - Cytomegalovirus (CMV) infection mainly affects endothelial cells of ocular vessels, optic nerve and the retina, resulting in direct or autoimmune damages, uveoretinitis and disturbed vision. The use of colloidal carriers for the intravitreal delivery of ganciclovir may prolong its residence in the eye, minimizing the opacification observed for macroscopic implants. The aim of this work was to evaluate the ocular toxicity induced by the prolonged presence of ganciclovir-loaded bovine serum albumin nanoparticles after their intravitreal injection. The intraocular disposition of these carriers was also studied by immunochemistry. Two weeks post-injection, a significant amount of nanoparticles remained in the vitreous cavity, mainly in a thin layer overlying the retina and in the area close to the blood aqueoUs barrier. Their prolonged residence in the eve seemed to be well tolerated and the histological evaluation of the retina, mainly the photoreceptor layer, and adjacent tissues revealed the absence of inflammatory reactions or alterations in the tissue architecture (i.e. cellular infiltrations or vascular inflammation). In addition, nanoparticles neither alter the expression and distribution of arrestin and rhodopsin autoantigens nor the mineralocorticoid receptor. In summary, the vision was not affected by autoimmune phenomena or alterations in the behavior of ophthalmic cells due to the intravitreal injection of these nanoparticles. PMID- 11922465 TI - Residual stress effects on fracture energies of cement-bone and cement-implant interfaces. AB - The effects of residual stresses, which are caused by the temperature difference arising after polymerisation of bone cement, on the fracture energies of cement bone and cement-implant interfaces have been examined by using both experimental and numerical works. Only fracture loads of the test specimen having interfacial cracks have been measured in the experimental stage. The values of fracture loads and temperature difference after polymerisation have been applied to finite element models of the test specimens to calculate critical J-integral values of these both interfaces in the numerical stage. In addition, fracture energies of bone and cement, have been obtained by experimentally, using three-point bending test method The results have shown that residual stresses can produce changes in the fracture energies of these bimaterial systems, especially in cement implant interface and J(Ic) values of interfaces are considerably smaller than the experimentally determined J(Ic) values of cement and bone. PMID- 11922466 TI - Influence of different surface modification treatments on poly(D,L-lactic acid) with silk fibroin and their effects on the culture of osteoblast in vitro. AB - The objective of this study was to investigate the efficiency of two treatments for poly(D,L-lactic acid) (PDLLA) surface modification using silk fibroin. one chemical treatment and one physical treatment: 1-(3-dimethylaminopropyl)-3 ethylcarbodiimide (WSC) and entrapment. The properties of control films, WSC modified and entrapment-treated PDLLA films were investigated by water contact angle measurement and electron spectroscopy for chemical analysis (ESCA). The water-contact angle measurement indicated the change of hydrophilicity and the ESCA analysis suggested that the modified PDLLA film using silk fibroin became enriched with nitrogen atoms. The biocompatibility of PDLLA film might be altered, which in turn would affect the functions of cells that were seeded on it. Therefore, attachment and proliferation of osteoblasts that were seeded on modified PDLLA films and control films were examined. Cell viability was evaluated by the MTT assay and differentiated cell function was assessed by measuring alkaline phosphatase activity. These results suggested that silk fibroin was used to modify PDLLA surface via WSC and that entrapment could improve the interactions between osteoblasts and PDLLA films. The entrapment treatment was more effective thin WSC treatment to accomplish the goal of surface modification. PMID- 11922467 TI - Apoptosis and cytokine release in human monocytes cultured on polystyrene and fibrinogen-coated polystyrene surfaces. AB - The effects of polystyrene (PS) material surface preadsorption with fibrinogen (3 mg/ml) and a low concentration of lipopolysaccharide (LPS; 10 ng/ml) and polystyrene particles (PS; 10(5)/ml) on human monocyte adhesion, viability and cytokine release were studied during 24h culture in vitro. LPS caused an upregulation of CD14 in adherent cells. In comparison with unstimulated cells on uncoated polystyrene surfaces, LPS did not alter the number of adherent cells but caused a markedly increased release of the proinflammatory cytokines (IL-1alpha and TNF-alpha) and the down-regulating IL-10. The expression of indicators of various stages of cell death, TdT, annexin-V, propidium iodide (PI) and lactate dehydrogenase (LDH), were unaltered, decreased, decreased and increased, respectively, after LPS stimulation. PS particles (3 microm psi) caused an increased DNA fragmentation but had a reduced proportion of annexin-V and PI positive cells in comparison with unstimulated cells on uncoated PS. In contrast, 1microm psi particles had a similar proportion of TdT, annexin-V and PI expressing cells as unstimulated controls. Cultures stimulated with particles (irrespective of size), had a similar concentration of proinflammatory cytokines as unstimulated controls, whereas a higher level of IL-10 was detected. Precoating of PS with fibrinogen revealed an enhanced cell adhesion and a concomitant reduction of CD14 expression. irrespective of stimulation with various agonists. The proportions of TdT, annexin-V and PI positive cells were unaltered or reduced on fibrinogen-coated PS in both unstimulated and agonist challenged cultures. However, depending on the presence and type of agonist, fibrinogen mediated either a markedly increased (LPS) or equivalent (particles and unstimulated) IL-1alpha and TNFalpha release. Further, in comparison with uncoated substrates, fibrinogen was associated with a reduction of IL-10 release, irrespective of the type of stimuli. These observations, using low concentrations of bacterial and material products, indicate that fibrinogen modulates cell material interactions and up- and down-regulates specific events depending on the nature/ type of immediate stimuli. PMID- 11922468 TI - Dexamethasone/PLGA microspheres for continuous delivery of an anti-inflammatory drug for implantable medical devices. AB - The purpose of this research was to develop polylactic-co-glycolic acid (PLGA) microspheres for continuous delivery of dexamethasone for over a 1-month period, in an effort to suppress the acute and chronic inflammatory reactions to implants such as biosensors, which interfere with their functionality. The microspheres were prepared using an oil-in-water emulsion technique. The oil phase was composed of 9:1 dichloromethane to methanol with dissolved PLGA and dexamethasone. Some microspheres were predegraded for 1 or 2 weeks. Ten percent of polyethylene glycol was added to the oil phase in alternative formulations to delay drug release. The in vitro release studies were performed in a constant temperature (37 C) warm room, in phosphate-buffered saline at sink conditions. Drug loading and release rates were determined by HPLC-UV analysis. The standard microsphere systems did not provide the desired release profile since, following an initial burst release, a delay of 2 weeks occurred prior to continuous drug release. Predegraded microspheres started to release dexamethasone immediately but the rate of release decreased after only 2 weeks. A mixed standard and predegraded microsphere system was used to avoid this delay and to provide continuous release of dexamethasone for 1 month. PMID- 11922469 TI - AFM and SFG studies of pHEMA-based hydrogel contact lens surfaces in saline solution: adhesion, friction, and the presence of non-crosslinked polymer chains at the surface. AB - The surfaces of two types of soft contact lenses neutral and ionic hydrogels- were characterized by atomic force microscopy (AFM) and sum-frequency-generation (SFG) vibrational spectroscopy. AFM measurements in saline solution showed that the presence of ionic functional groups at the surface lowered the friction and adhesion to a hydrophobic polystyrene tip. This was attributed to the specific interactions of water and the molecular orientation of hydrogel chains at the surface. Friction and adhesion behavior also revealed the presence of domains of non-crosslinked polymer chains at the lens surface. SFG showed that the lens surface became partially dehydrated upon exposure to air. On this partially dehydrated lens surface, the non-crosslinked domains exhibited low friction and adhesion in AFM. Fully hydrated in saline solution, the non-crosslinked domains extended more than tens of nanometers into solution and were mobile. PMID- 11922470 TI - Mucoadhesive polymers with immobilized proteinase inhibitors for oral administration of protein drugs. AB - A new approach to overcome the degradation of protein drugs by proteolytic enzymes and their targeting to the blood through the digestive apparatus was developed. The approach is based on the immobilization of drugs into the polymeric hydrogel containing glycoprotein--ovomucoid from duck egg whites. This glycoprotein inhibits the activity of proteolytic enzymes and acts as a biospecific ligand to lectins on the walls of the gastrointestinal tract. PMID- 11922471 TI - Structural evolution of sol-gel-derived hydroxyapatite. AB - Structural evolution upon transformation of sol to gel, and gel to final ceramic during the synthesis of hydroxyapatite is investigated using Fourier transform infrared (FTIR) analysis, X-ray diffraction (XRD), thermal behavior (DTA and TGA), and electron microscopy examination (SEM/TEM). The sol was first thermally aged at 45 C for various time periods up to 120 min. The colloidal sol, which may have an oligomeric structure, was relatively stable against coagulation. Upon drying, the sol particles consolidated into dry gel through van der Waals attraction, and showed X-ray amorphous phosphate structure. The solid gels showed a particulate microstructure, composed of primary particles of about 8-10 nm in diameter. The amorphous gel transformed into crystalline apatite at temperatures > 300 C. The calcined gels showed a nano-scale microstructure, with grains of 20 50 nm in diameter. Through an appropriate heat treatment between 300 and 400d degrees C. the apatite prepared using current process exhibits a nano-scale, low crystallinity, carbonated apatitic structure, which closely resembles that of human bone apatite. PMID- 11922472 TI - TCP is hardly resorbed and not osteoconductive in a non-loading calvarial model. AB - Tricalciumphosphate (TCP) has been used as a ceramic bone substitute material in the orthopedic field as well as in craniofacial surgery. Some controversies exist concerning the osteoconductive potential of this material in different implantation sites. This study was designed to evaluate the biological response of calvarial bone towards TCP granules under non-loading conditions to assess the potential of TCP as a biodegredable and osteoconductive bone substitue material for the cranial vault. Full-thickness non-critical size defects were made bilaterally in the calvaria of 21 adult Wistar rats. One side was filled by TCP granules, the contralateral side was left empty and used as a control. Animals were sacrified in defined time intervals up to 6 months. Bone regeneration was analyzed with special respect toward the micromorphological and microanalytical features of the material-bone interaction by electron microscopy and electron diffraction analysis. Histologic examination revealed no TCP degradation even after 6 months of implantation. In contrast, a nearly complete bone regeneration of control defects was found after 6 months. At all times TCP was surrounded by a thin fibrous layer without presence of osteoblasts and features of regular mineralization. As far as degradation and substitution are concerned, TCP is a less favourable material tinder conditions of non-loading. PMID- 11922473 TI - Analysis of a phosphorylcholine-based polymer coating on a coronary stent pre- and post-implantation. AB - There has been a move towards surface treatments for metallic coronary stents in an effort to improve their compatibility within the body and to provide a vehicle for the delivery of therapeutics. The Biodiv Ysio range of stents is characterised by a biocompatible coating comprised of a crosslinked phosphorylcholine (PC)-based polymer. In addition to a review of some of the data collected to support safety and efficacy of this device, this paper also describes a number of techniques that have been employed to both visualise and quantify the coating on the stent. Explantation of both coated and uncoated stents from porcine coronary arteries revealed that both coated and uncoated stents were >90% endothelialised after 5 days. Typical histological analysis of stented vessel sections after 4 and 12 weeks implantation showed the presence of cell types characteristic of the inflammatory response associated with the trauma caused by stent placement, with no evidence for any additional coating-related adverse inflammatory sequelae. Finally, it was demonstrated by AFM and SEM that both the thickness and force required to remove the coating were essentially unchanged after 6 months implantation. Thus, both the long-term stability and relative biological inertness of the coating has been confirmed in vivo, supporting its use as a vehicle for local drug delivery. PMID- 11922474 TI - Interactions between resin monomers and commercial composite resins with human saliva derived esterases. AB - Cholesterol esterase (CE) and pseudocholinesterase (PCE) have been reported to degrade commercial and model composite resins containing bisphenylglycidyl dimethacrylate (BisGMA), triethylene glycol dimethacrylate (TEGDMA) or the latter in combination with urethane modified BisGMA monomer systems. In addition, human saliva has been shown to contain esterase like activities similar to CE and PCE. Hence, it was the aim of the current study to determine to what extent human saliva could degrade two common commercial composite resins (Z250 from 3M Inc. and Spectrum TPH from L.D. Caulk) which contain the above monomer systems. Saliva samples from different volunteers were collected, processed, pooled, and freeze dried. TEGDMA and BisGMA monomers were incubated with human saliva derived esterase activity (HSDEA) and their respective hydrolysis was monitored using high performance liquid chromatography (HPLC). Both monomers were completely hydrolyzed within 25 h by HSDEA. Photopolymerized composites were incubated with buffer or human saliva (pH 7.0 and 37 C) for 2, 8 and 16 days. The incubation solutions were analyzed using HPLC and mass spectrometry. Surface morphology characterization was carried out using scanning electron microscopy. Upon biodegradation, the Z250 composite yielded higher amounts of BisGMA and TEGDMA related products relative to the TPH composite. However, there were higher amounts of ethoxylated bis-phenol A released from the TPH material. In terms of total mass of products released, human saliva demonstrated a greater ability to degrade Z250. In summary, HSDEA has been shown to contain esterase activities that can readily catalyze the biodegradation of current commercial composite resins. PMID- 11922475 TI - In vivo degradation and biocompatibility of a new class of alternate poly(ester anhydrides) based on aliphatic and aromatic diacids. AB - The degradation, tissue compatibility, and toxicology of a novel class of alternate poly(ester-anhydrides) were assessed in rats. It was observed that the degradation rate of the polymers in vivo was slower than that in vitro. In addition, erosion and intact zone were observed for all the polymers. IR and SEM analysis of the outer erosion and inner intact zone revealed that the outer zone degraded more rapidly than the inner zone. Such results were similar to that in vitro. All the studied poly(ester-anhydrides) produced mild inflammatory reactions and tissue encapsulation by layers of fibroblastic cells in vivo. Observation of liver and kidney tissue by light microscopy suggested the hydrolytic products of the studied poly(ester-anhydrides) had no harmful effects on the normal tissue/organs. In addition, the polymer and the breakdown products were found to be non-mutagenic by examination of micronucleus in bone marrow. PMID- 11922476 TI - Hydroxyapatite deposition by alternating soaking technique on poly(vinyl alcohol) coated polyethylene films. AB - A poly(vinyl alcohol) (PVA)-coating on polyethylene films, prepared by repetitive adsorption/drying in an aqueous PVA solution, accelerated hydroxyapatite (HAp) deposition by an altemate soaking in aqueous solutions containing Ca2+ and PO4(3 ) ions. X-ray photoelectron spectra of the surface of the HAp-deposited film showed the presence of calcium and phosphorus of a suitable peak ratio for HAp formation. X-ray diffraction analyses also revealed peaks corresponding to HAp. Scanning electron microscopic observation showed the surface of the HAp layer to be smooth, with nano-ordered dotted threads in networks. A simple PVA coating on a surface will serve as a novel system for accelerated HAp formation via alternating soaking. PMID- 11922477 TI - Rat osteoblast functions on the o-carboxymethyl chitosan-modified poly(D,L-lactic acid) surface. AB - In this study, the functions of rat osteoblasts on o-carboxymethyl chitosan modified poly(D,L-lactic acid) (PDLLA) films were investigated in vitro. The surface characterization was measured by contact angle and electron spectroscopy for chemical analysis (ESCA). Cell adhesion and proliferation were used to assess cell behavior on the modified surface and control. The MTT assay was used to determined cell viability and alkaline phosphatase (ALP) activity was performed to evaluate differentiated cell function. Compared to the control films, cell adhesion of osteoblasts on o-carboxymethyl chitosan-modified PDLLA films was significantly higher (p < 0.05) after 6 and 8 h culture, and osteoblast proliferation was also significantly higher (p < 0.01) between 4 and 7 days. The MTT assay suggested cell viability of osteoblasts cultured on o-carboxymethyl chitosan modified PDLLA films was significantly greater (p < 0.05) than that seeded on control one, and the ALP activity of cells cultured on modified PDLLA films was significantly higher (p < 0.01) than that found on control. These results give the first evidence that o-carboxymethyl chitosan could be used to modify PDLLA surface for improving biocompatibility. PMID- 11922478 TI - Effect of preparation conditions on properties and permeability of chitosan sodium hexametaphosphate capsules. AB - Capsules were obtained by interpolymer complexation between chitosan (polycation) and sodium hexametaphosphate (SMP, oligoanion). The effect of the preparation conditions on the capsule characteristics was evaluated. Specifically, the influence of variables such as pH, ionic strength, reagent concentration, and additives on the capsule permeability properties was investigated using dextran as a model permeant. The capsule membrane permeability was found to increase by decreasing the chitosan/SMP ratio as well as adding mannitol to the oligoanion recipient bath. Increasing the ionic strength or the pH of the initial chitosan solution was also found to enhance the membrane permeability, moving the membrane exclusion limit to higher values. Generally, the capsules prepared tinder all tested conditions had a relatively low permeability which rarely exceeded a molecular cut-off of 40 kD based on dextran standards. Furthermore, the diffusion rate showed a strong temporal dependence, indicating that the capsules prepared under various conditions exhibit different apparent pore size densities on the surface. The results indicated that, in order to obtain the desired capsule mass transfer properties, the preparation conditions should be carefully considered and adjusted. Adding a polyol as well as low salt amount (less than 0.15%) is preferable as a means of modulating the diffusion characteristics, without disturbing the capsule mechanical stability. PMID- 11922479 TI - Tailor-made core-shell nanospheres for antisense oligonucleotide delivery: IV. Adsorption/release behaviour. AB - The adsorption/release behaviour of oligodeoxynucleotides (ODNs) on double functional core-shell polymethylmethacrylate nanospheres, with a narrow size distribution, is described. The outer shell consists of alkyl or glycolic chains containing permanently-charged quaternary ammonium groups. Ion pair formation between negatively-charged ODN phosphate groups and positively-charged groups, present on the nanosphere surface, is the main mechanism of interaction. The amount of adsorbed ODN depends on both the ODN concentration and the nanosphere surface charge density. An adsorption-induced swelling mechanism is proposed in which a modification of the charged diffuse layer around the nanospheres increases the ODN binding site accessibility with increasing ODN concentration. Adsorption on the nanosphere surface prevents serum degradation of the ODNs. ODN release is negligible in the presence of culture medium but occurs gradually in the presence of serum. No significant cytotoxicity of the free nanoparticles was found in PBMC and CEM cells after 24 h at ODN concentrations required for antisense activity. PMID- 11922480 TI - Adsorption studies of human serum albumin, human gamma-globulins, and human fibrinogen on the surface of p(S/PGL) microspheres. AB - Adsorption of human serum albumin (HSA), human gamma-globulins (gammaG), and human fibrinogen (Fb) onto the surface of poly(styrene/alpha-t-butoxy-omega vinylbenzyl-polyglycidol) microspheres (P(S/PGL)) with controlled fraction of polyglycidol in the interfacial layer was investigated. The microspheres were synthesized by the emulsifier-free radical copolymerization of styrene and alpha t-butoxy-omega-vinylbenzyl-polyglycidol macromonomer (PGL). Macromonomers with number average molecular weights Mn = 950 and 2,700 were used in the syntheses. Fraction of polyglycidol in the microsphere surface layer was varied from 0.22 to 0.44, depending on the composition of the monomer feed. It was found that the maximal surface concentration of adsorbed proteins and the equilibrium constant of protein adsorption decreased with increased fraction of polyglycidol in the microsphere surface layer. For microspheres with the highest fraction of polyglycidol at the surface the maximal surface protein concentration was c. ten times lower and the adsorption equilibrium constant was c. one hundred times lower than for the reference polystyrene microspheres. The dependence of maximal surface concentration of adsorbed proteins on the fraction of polyglycidol in the particle interfacial layer indicated random distribution of polyglycidol chains without formation of polyglycidol and polystyrene patches at the microspheres surface. PMID- 11922481 TI - Neutrophil phagocytosis following inoculation of Salmonella choleraesuis into swine. AB - Neutrophils are an important mediator of host defence, especially in early stages of infection. A major function of neutrophils is the uptake and killing of invading microbes. Little is known about the effect of neutrophil activity on the pathogenesis and development of the carrier state in swine following infection with Salmonella choleraesuis. A human whole-blood microassay using flow cytometry was modified to measure the effect of S. choleraesuis infection in vivo on the rate of ingestion, or rate of uptake, of homologous bacteria by porcine neutrophils. Pigs were inoculated intranasally with 5-8 x 10(8) CFU S. choleraesuis and blood was collected in heparinized tubes at -5, 0, 1, 2, 3 and 4 days post inoculation (PI). Heat-killed S. choleraesuis were labelled with fluorescein isothiocyanate and incubated for various times with diluted whole blood. Red blood cells were lysed, external non-phagocytized bacteria were quenched with a commercially available lysing solution, and fluorescence from internalized bacteria labelled with fluorescein isothiocyanate was detected by flow cytometry. The rate of uptake by neutrophils did not increase until 2 days PI and then remained elevated to 4 days PI. The minimal uptake of S. choleraesuis early after exposure to these organisms may provide an opportunity for the pathogen to colonize and/or replicate to levels that facilitate establishment of a carrier state or clinical infection in swine. PMID- 11922482 TI - Measurement of cortisol metabolites in faeces of ruminants. AB - Twenty-one metabolites were detected in faecal samples collected after infusion of (14C)cortisol into the jugular vein of sheep. Using high-performance liquid chromatography/radiometric analysis plus mass spectrometry. One group of metabolites had molecular weights of between 302 and 308, and another group of 350, which indicates that the substances have a C19O3 or a C21O4 structure. Therefore, an enzyme immunoassay against 5beta-androstane-3alpha-ol-11,17-dione 17-CMO:BSA was established. Faecal samples were collected from 10 cows immediately after transport and then during a course in which non-invasive diagnostic procedures were being taught (course 1). For comparison, faeces were sampled from another 5 cows that were being used for teaching invasive procedures (course 2). Six cows from a university farm served as controls. In the animals used in course 1, the highest concentrations of cortisol metabolites were measured immediately after transport to the university (median value: 2.2 micromol/kg faeces). During the first 5 days at the university, the concentrations decreased to 0.52 micromol/kg (median) and remained at this level during the rest of the course. The median concentration in the samples that were taken during coursc 2 (collected about 2 months after transport) was 0.48 micromol/kg. There was no significant difference in the excretion of cortisol metabolites between these cows and the controls. We conclude from these data that, using the enzyme immunoassay against 5beta-androstane-3alpha-ol-11,17-dione 17-CMO, we were able to detect transport/novel environment stress but not the potential disturbance that cows experience during diagnostic procedures. PMID- 11922483 TI - Pharmacokinetics of pefloxacin in goats after intravenous or oral administration. AB - The plasma concentrations and pharmacokinetics of the fluoroquinolone antimicrobial agent pefloxacin, following the administration of a single intravenous (10 mg/kg) or oral (20 mg/kg) dose, were investigated in healthy female goats. The antimicrobial activity in plasma was measured at predetermined times after drug administration by an agar well diffusion microbiological assay, using Escherichia coli (ATCC 25922) as the test organism. Concentrations of the drug > or = 0.25 microg/ml were maintained in plasma for up to 6 and 10 h after intravenous (i.v.) or oral administration of pefloxacin, respectively. The concentration time data for pefloxacin in plasma after i.v. or oral administration conformed to two- and one-compartment open models, respectively. Plasma pefloxacin concentrations decreased rapidly during the initial phase after i.v. injection, with a distribution half-life (t(1/2alpha)) of 0.10 +/- 0.01 h. The terminal phase had a half-life (t(1/2beta)) of 1.12 +/- 0.21 h. The volume of distribution at steady state (Vdss), mean residence time (MRT) and total systemic clearance (ClB) of pefloxacin were 1.08 +/- 0.09 L/kg, 1.39 +/- 0.23 h and 821 +/ 88 (ml/h)/kg, respectively. Following oral administration of pefloxacin, the maximum concentration in the plasma (Cmax) was 2.22 +/- 0.48 microg/ml and the interval from administration until maximum concentration (tmax) was 2.3 +/- 0.7 h. The absorption half-life (t(1/2ka)) mean absorption time (MAT) and elimination half-life of pefloxacin were 0.82 +/- 0.40, 4.2 +/- 1.0 and 2.91 +/- 0.50 h, respectively. The oral bioavailability of pefloxacin was 42% +/- 5.8%. On the basis of the pharmacokinetic data, a dosage regimen of 20 mg/kg, i.v. at 8 h intervals or orally twice daily, is suggested for treating infections caused by drug-sensitive pathogens in goats. PMID- 11922484 TI - The effect of melatonin on the secretion of progesterone in sheep and on the development of ovine embryos in vitro. AB - Two experiments were carried out in order to determine whether melatonin can improve secretion of progesterone in vivo, and its effect on embryonic development in vitro. In the first experiment, blood samples were collected from 5 ewes at 15 min intervals for 2 h at 7 and 10 days after withdrawal of progestagen pessaries. The first hour constituted a control period, which ended with an intravenous administration of 3 microg/(kg bw)(0.75) melatonin. All the ewes on day 7 and three of the ewes on day 10 showed a progesterone response to melatonin challenge, defined as an increase in the plasma progesterone concentration in at least two consecutive samples during the post-treatment period above the mean+2SD of the values in the pre-treatment period. A paired t test revealed a significant effect of melatonin on the overall plasma progesterone concentrations before and after the challenge, both on day 7 (pre, 0.61 +/- 0.11; post, 0.73 +/- 0.13 ng/ml; p<0.01) and day 10 (pre, 1.16 +/- 0.19; post, 1.30 +/- 0.20 ng/ml; p<0.05). Ninety-one thawed embryos (46 morulae and 45 blastocysts) were used in the second experiment, being cultured with or without 1 microg/ml melatonin. If the embryos were blastocysts when the culture started. melatonin increased the percentage that had hatched after 24 h of culture (p<0.01), and there was a lower percentage of degenerated embryos at the end of the incubation period (p<0.05). It may be concluded that melatonin treatment in sheep can increase both fertility and prolificacy by improving luteal function and embryonic survival. PMID- 11922485 TI - Ruminal metabolism in sheep of saponins from Yucca schidigera. AB - Fifty-five mg per kg live weight of crystallized Yucca schidigera saponins, corresponding to 26 mg/kg live weight of sapogenins, was given daily intraruminally to two lambs for 11 consecutive days. Neither of the lambs showed any sign of toxicity throughout the experimental period. One lamb was killed 5 h after the last dose and GC-MS analysis of the free and conjugated sapogenin content samples of liver, and of the contents of the rumen, omasum, abomasum, duodenum, jejunum, ileum, colon and rectum, of faecal samples collected before dosing started, and of parts of the administered Yucca saponin were performed. The Yucca material contained mainly sarsasapogenin and smilagenin saponins. Ingested saponins were quickly hydrolysed in the rumen to free sapogenins and, in part, epimerized at C-3 to afford episapogenins. The absorption of free sapogenins appeared to occur in the jejunum. The metabolism of Yucca saponins was identical to that of Narthecium ossifragum saponins, and it is suggested that Yucca saponins could replace N. ossifragum saponins for toxicity studies on the latter plant. PMID- 11922486 TI - The effect of Brucella abortus on hydrogen peroxide and nitric oxide production by bovine polymorphonuclear cells. AB - The effect of Brucella on the generation of microbicidal reactive oxygen and nitrogen metabolites by bovine peripheral polymorphonuclear cells (PMNs) was investigated. The PMNs were recovered from the peripheral blood of control calves and experimental calves previously vaccinated against brucellosis. Significantly larger quantities of NO and H2O2 were generated by PMNs from control and experimental calves following activation by heat-killed whole cells or outer membrane protein of Brucella abortus than by non-activated cells (p<0.05-0.01). In contrast, generation of H2O2 and NO decreased when PMNs were exposed to the lipopolysaccharide of Brucella. However, the generation of H2O2 and NO by activated PMNs from the control and experimental calves did not differ significantly. PMID- 11922487 TI - Tic disorders: when habit forming neural systems form habits of their own? AB - Tourette syndrome (TS), obsessive-compulsive disorder (OCD) and related conditions are prevalent disorders affecting as many as 0.3-3% of the population. They are frequently chronic and can be associated with marked impairment and disability. Although clinical care has improved over the past decade, a significant number of patients fail to respond adequately or experience intolerable side effects. The etiology of these disorders is unknown. Compelling evidence suggests that the vulnerability to develop TS and OCD is mediated by both genetic and environmental factors, and that neural systems located in the basal ganglia and functionally related brain structures are involved in their pathogenesis. Based on explicit models of pathogenesis for TS and OCD and building on work accomplished over the past two decades, an array of clinical, neuropsychological, genetic, neuroimaging, epidemiological neurobiological, and treatment studies have been completed or are underway at the Child Study Center at Yale University. A multidisciplinary team of investigators has joined forces to test specific hypotheses through the integration and translation of basic and clinical neuroscience research. All subjects have been studied using identical clinical, neuropsychological, genetic, neurobiological, and pharmacological techniques. Current conceptualizations of TS have been shaped by advances in clinical phenomenology, genetics, systems neuroscience and the emerging understanding of the role of the basal ganglia in implicit learning and habit formation, neuroimmunology and psychopharmacology. An appreciation of the premonitory urges that precede tics and temporal dynamics of tics have provided useful viewpoints from which to regard the natural history of TS. While the long term outcome of TS can be relatively benign, the presence of comorbid conditions such as attention deficit/hyperactivity disorder (ADHD), OCD or a major affective disorder can have lasting untoward consequences. The identification of susceptibility genes in TS will doubtless point in new therapeutic directions for treatment, as will the characterization of the putative autoimmune mechanisms active in subgroup of patients. Continued success in functional in vivo neuroimaging studies will lead to the targeting of specific brain circuits for more intensive study. Although ideal anti-tic therapies are not available, recently completed clinical trials with alpha-adrenergic agents and atypical neuroleptics are encouraging. Given these developments, TS can be considered a model disorder to study the dynamic interplay of genetic vulnerabilities, epigenetic events, and neurobiological systems active during early brain development. It is likely that the research paradigms utilized in these studies and many of the empirical findings resulting from them, will be relevant to other disorders of childhood onset and to our understanding of normal development. PMID- 11922489 TI - Pulse wave velocity: is it a practical diagnostic modality? PMID- 11922488 TI - Sequential tonometry as a practical method to estimate truncal pulse wave velocity. AB - BACKGROUND: Pulse wave velocity (PWV) is a surrogate marker of arteriosclerosis. It can be measured by recording pressure or flow wave signals at two recording sites of an arterial segment simultaneously or sequentially. However, it is unknown how the values of pulse wave velocity derived from various techniques can be compared. Thus, the aims of the present study were to investigate whether different techniques result in different values of PWV. METHODS: We measured aortic PWV from 101 uremic and non-uremic patients using both applanation tonometry and Doppler velocimetry by sequential method. To evaluate whether there is a difference between the sequentially and simultaneously derived PWV, simultaneous method using 2 tonometer or 2 Doppler probes was also applied in 50 and 23 subjects, respectively. RESULTS: There was no significant difference between PWV derived from sequential and simultaneous tonometry (933+/-310 cm/s and 919+/-301 cm/s,p = 0.09). Likewise, there was no significant difference between PWV derived from sequential and simultaneous velocimetry (778+/-241 cm/s and 761+/-205 cm/s,p = 0.35). However, PWV determined by sequential tonometry was significantly greater than (949+/-315 cm/s and 735+/-208 cm/s, respectively,p < 0.001), yet also significantly correlated with that determined by sequential velocimetry (r = 0.83, p < 0.001). CONCLUSIONS: Sequential applanation tonometry is a practical approach to measure PWV in view of technical convenience and the cost of equipment and manpower. Furthermore, we should be cautious in interpreting PWV derived from different methodologies as greater PWV value might be obtained by applanation tonometry than by Doppler velocimetry. PMID- 11922491 TI - Retinopathy of prematurity: screening, incidence and risk factor analysis. PMID- 11922490 TI - Retinopathy of prematurity: screening, incidence and risk factors analysis. AB - BACKGROUND: The sequela of retinopathy of prematurity (ROP) is an important cause of infant blindness. This study was designed to screen the high-risk premature infants and investigate the incidence and risk factors associated with the development of ROP. METHODS: From October 1997 to October 1998, all premature infants with birth body weight (BBW) less than 2000 gm or gestational age (GA) less than 36 weeks were enrolled and underwent ophthalmologic examination at 4 to 6 weeks of age at Taipei Veterans General Hospital. The perinatal variables were analyzed to evaluate their correlation with the development of ROP. RESULTS: In totally 108 premature infants, the incidence for development of ROP was 25% (27 in 108 patients). The threshold ROP occurred in 15 eyes (7%). The average BBW and GA (1267+/-341 gm and 29.7+/-2.7 weeks) were significantly lower in ROP group than in the non-ROP group (1703+/-368 gm and 32.3+/-2.2 weeks). The artificial ventilation for more than 5 days, chronic lung disease and periventricular leukomalacia were significant risk factors associated with highest rate of ROP. The respiratory distress syndrome, intraventricular hemorrhage, congenital heart disease and sepsis were significant risk factors accompanied by moderate rate of ROP. CONCLUSION: Low birth body weight and young gestational age are the most important risk factors in the development of ROP. The analysis of risk factors will be helpful in understanding and prediction of ROP formation in high-risk neonates. The timely clinical screening retina examination of high-risk premature infants is important to prevent the development of advanced ROP. PMID- 11922493 TI - Rectal administration of misoprostol for the management of retained placenta--a preliminary report. AB - BACKGROUND: Retained placenta is one of the serious complications of childbirth, and misoprostol is known to be a potent uterotonic agent. Therefore, we proposed that rectal misoprostol also may facilitate placental separation in women with retained placenta by its ability to increase uterine contractility. METHODS: The placenta was diagnosed as retained if it was not expelled within 40 minutes after vaginal birth. Then, 800 microg of misoprostol was inserted rectally and the patient observed thereafter. RESULTS: A total of 18 parturients who had retention of the placenta were studied; all the placentas were spontaneously expelled within 35 minutes. The side effects involved included nausea 17%, vomiting 11%, diarrhea 22%, shivering 33%, and pelvic cramping pain 44%. All these discomforts resolved within 24 hours. CONCLUSIONS: Our study demonstrated that misoprostol per rectum is a safe and effective technique and may be a useful alternative to manual removal of retained placentas. PMID- 11922492 TI - Effects of verapamil on coronary vascular resistance in rabbits: measurement with pulsed Doppler velocimetry. AB - BACKGROUND: Verapamil is an effective vasodilator. The purpose of this study was to investigate the in vivo effect of verapamil on coronary blood flow velocity and vascular resistance in anesthetized, open-chest rabbits. METHODS: Twenty-one male New Zealand white rabbits were anesthetized, and a 3-mm suction-type pulsed Doppler velocimeter probe was applied to the proximal part of the left anterior descending coronary artery after median sternotomy. The rabbits received intravenous bolus infusion of 4 different doses of verapamil (0.01 mg/kg, n = 5; 0.1 mg/kg, n = 5; 1 mg/kg, n = 5, and 10 mg/kg, n = 6). The percent changes in coronary blood flow velocity and coronary vascular resistance were examined. RESULTS: There was 10.0+/-1.6% increase in coronary blood flow (CBF) and 12.5+/ 1.9% reduction in coronary vascular resistance (CVR) after infusion of 0.01 mg/kg of verapamil. The CBF increased 23.0+/-9.5% and CVR decreased 24.2+/-5.2% after infusion of 0.1 mg/kg of verapamil. Infusion of 1 mg/kg of verapamil induced 34.8+/-10.5% increase in CBF and 32.6+/-2.5% reduction in CVR. The CBF increased 41.1+/-14.8% and CVR decreased 45.1+/-5.4% after infusion of 10 mg/kg of verapamil. CONCLUSIONS: Compared with baseline condition, all doses of verapamil increased coronary blood flow velocity and decreased coronary vascular resistance significantly in anesthetized, open-chest rabbits. PMID- 11922494 TI - A progressive growing inflammatory pseudotumor of the liver. AB - Inflammatory pseudotumors of the liver are very rare, and their etiology and pathogenesis remain unclear. The diagnosis is often difficult to make because these masses often mimic other lesions such as primary neoplasms, metastases or liver abscesses. Herein, we report a rare case of progressive growing hepatic pseudotumor in a 47-year-old man. The patient presented with body weight loss and general malaise. A series of radiological examinations showed the progressive growth of the hepatic tumor from 3.5 cm to 10.0 cm in diameter within 8 months. He underwent a right lobectomy of the liver, and the final diagnosis was proven by pathology. There were no complications in the post-operative course. PMID- 11922495 TI - Hemoclip-assisted endoscopic polypectomy of large superior duodenal angle polyp using a needle knife. AB - Endoscopic polypectomy of a large polyp can be difficult due to inability to snare the polyp. The difficulty may increase when the polyp is located at turning corner of the bowel. We presented a case of a 3 cm-sized large pedunculated polyp located at the superior duodenal angle that was not amenable to conventional snare polypectomy, but was instead successfully resected by hemoclip-assisted and needle knife method. Such experience has not been reported in the English literature. PMID- 11922496 TI - Propylthiouracil-induced hemolytic anemia. AB - Propylthiouracil (PTU)-induced hemolytic anemia is extremely rare. We reported a case of Graves' disease with these unusual clinical manifestations. A 41-year-old female presented with recurrent attacks of severe hemolytic anemia after PTU therapy. Sugar water test and erythrocytes osmotic fragility test revealed no cellular membrane defect of red blood cells. Antinuclear antibody, direct and indirect Coombs' tests were all negative and glucose-6-phosphate dehydrogenase activity was also within normal limits. PTU was not discontinued promptly due to unrecognizableness of such a rare case until two months later with recurrent attacks of severe hemolytic anemia. 1-131 therapy was performed on suspicion of related hemolytic anemia. Unfortunately, challenge of PTU occurred incidentally after discontinuation of PTU followed by severe hemolytic anemia. The diagnosis of PTU-induced hemolytic anemia was established thereafter. A MEDLINE search revealed only one such case reported in English literature. This is the first case report in Taiwan. It should be kept in mind that hemolytic anemia may be a rare complication of PTU therapy. PMID- 11922497 TI - Carvedilol reverses elevated pulmonary vascular resistance in a child with dilated cardiomyopathy. AB - There is little experience using beta-blocker therapy for the management of congestive heart failure (CHF) in children. We present the case of a child with idiopathic dilated cardiomyopathy and CHF, referred for cardiac transplantation, in whom carvedilol reversed elevated pulmonary vascular resistance. PMID- 11922499 TI - Left pulmonary venous stenosis with left isomerism heart: a case report of intravascular ultrasound imaging. PMID- 11922498 TI - Persistence of the right valve of the sinus venosus resulting in an unusually prominent Chiari network remnant mimicking cor triatriatum dexter. PMID- 11922500 TI - Aortico-cameral communication from left sinus Valsalva aneurysm to right atrium via a tortuous tunnel with aneurysmal dilatation. PMID- 11922501 TI - About congenital polyvalvular disease. PMID- 11922502 TI - Circulatory arrest and renal function in open-heart surgery on infants. AB - This study was designed to evaluate the impact of circulatory arrest on renal function in open-heart surgery on infants. Renal function was described by diuresis, urine/plasma creatinine ratio, creatinine clearance, urinary albumin, and N-acetyl-beta-D-glucosaminidase measurements. Seven patients who underwent circulatory arrest (group 1) were compared with 37 patients operated on with cardiopulmonary bypass without circulatory arrest (group 2). In group 1, bypass time was 164 minutes and the lowest body temperature was 25.6 degrees C (median), compared with 106 minutes and 31.3 degrees C in group 2 (p < 0.05). Although diuresis and creatinine clearance revealed no differences between the groups, urine measurements, which had normal values before cardiopulmonary bypass, increased during reperfusion to 58.6 (range 16.2-75.5) mg gCrea(-1) albumin and to 14.8 (range 1.6-21.8) U gCrea(-1) N-acetyl-beta-D-glucosaminidase in group 1, compared with 8.1 (range 0-127.7) mg gCrea(-1) and 1.9 (range 0-47.8) U gCrea(-1) in group 2 (p < 0.05). Thus, deep hypothermic circulatory arrest (DHCA) subjected the kidney to ischemia reperfusion injury. Although the findings are mild and do not indicate severe ischemic renal damage, potential renal damage by DHCA should be taken into account when planning surgical procedures for congenital heart disease patients with additional risks of acute renal failure. PMID- 11922503 TI - Autosomal dominant myocardial disease diagnosed by fetal presentation of proband with an aneurysm of the muscular interventricular septum. AB - We describe an inherited form of a disorder in which four patients spanning three generations were affected with congenital myocardial disease. The youngest member of the family, diagnosed as a fetus with a large aneurysm of the muscular interventricular septum, demonstrates an antenatal pathogenic process. Study of the specific findings in each patient suggests that congenital aneurysms of the muscular interventricular septum may be in some way associated with specific developmental pathways of the heart. We believe that screening family members of patients with muscular interventricular septal aneurysms may be indicated to assess for silent myocardial disease. PMID- 11922504 TI - Growth after neonatal arterial switch operation for D-transposition of the great arteries. AB - The objective of this study was to evaluate growth in infants with d transposition of the great arteries (d-TGA) who underwent switch operation (anatomical correction) in the early neonatal period. Growth data (at birth and 1, 3, 6, 12, and 24 months of age) were retrieved in 23 infants with d-TGA who survived the operation without major complications. Measures were transformed into z scores and compared with normative data for age. Following surgery, infants showed low z scores for weight, height, and head circumference. Weight and height showed a trend to reach normal ranges between 6 and 12 months of age, and the z scores were positive at 24 months. z scores for head circumference were still negative at 24 months of age. No dietary interventions were implemented during this period, and the infants were fed their own mothers' milk or, before discharge, high-calorie, preterm formula. Following anatomical correction for d TGA in the early neonatal period, infant growth shows a characteristic pattern, with parameters normalized by 12 months of age in most cases. Head growth shows a different pattern, although gross motor development seems unaffected. PMID- 11922505 TI - Around PediHeart: cardiomyopathy and renal failure. PMID- 11922506 TI - Comparison of atrial septal defect closure using amplatzer septal occluder with surgery. AB - Our study reports the results of a comparison of closure of atrial septal defect (ASD) surgically with transcatheter closure using the Amplatzer septal occluder. Patients having an ASD and a surgical closure or transcatheter closure between January 1999 and July 2000 were selected. There were 103 patients who had ASD. All 64 patients in group 1 (surgery) had a successful operation, with only 2 patients with a mild residual shunt. There were 39 patients enrolled for transcatheter closure of the ASD (group 2). Four patients were excluded initially. The median age for group 1 was 25 years (range 2.3-64 years) compared to 11.7 years (range 2-69 years) in group 2 (p = 0.035). In group 1, the mean ASD diameter measured was 28.4 +/- 10.2 mm compared to 23.4 +/- 5.7 mm in group 2 (p = 0.003). In 29 patients, devices were deployed with sizes from 10 mm to 30 mm (median 24 mm). Three patients were excluded because a larger device (>30 mm) was not available and devices were not successfully deployed in another 3 patients. One patient had a device embolized into the right ventricle (surgical removal and closure of the ASD). Complications occurred in 13 patients in group 1 and 4 patients in group 2. Complete occlusion occurred in 27 of 28 group 2 patients (96.4%) during the follow-up period (10.2 +/- 5.4 months). The Amplatzer septal occluder is a new device for closure of different-sized ASDs. The intermediate term follow-up demonstrated excellent closure results. The benefit for each patient was demonstrated in less morbidity and less time spent in the hospital. PMID- 11922507 TI - Studies of magnesium in congenital long QT syndrome. AB - We studied the role of magnesium (Mg) in congenital long QT syndrome (LQTS). Twenty-two congenital LQTS patients and 30 control subjects were included in this study. We measured serum Mg (SMg) level and Mg retention (MgR) level, and evaluated the role of Mg (a high MgR level reflects Mg deficiency in the body). The influence of intravenous Mg infusion on Mg level was evaluated. Relatively low SMg level and high MgR level (LQTS:control = 53:33%, p < 0.01) were recognized in congenital LQTS patients, but there was an overlap with controls. Mg supplementation did not shorten QT interval and there was no significant correlation between Mg levels and QTc interval. Patients with syncopal history showed a higher MgR level (syncope (+):syncope (-) = 70:46%, p < 0.01) and intravenous Mg infusion improved Mg deficiency. These results suggest that some (not all) congenital LQTS patients are in a Mg-deficient state, which may be associated with syncope, and Mg supplementation may prevent recurrent syncope in these patients. Because there are several subtypes of congenital LQTS, perhaps with genetic testing Mg deficiency may be identified as a significant cofactor in some forms, whereas in other forms it is not relevant. PMID- 11922508 TI - The modified Blalock-Taussig shunt: a 6-year experience from a developing country. AB - Since 1992 we have performed the modified Blalock-Taussig shunt (MBTS) for cyanotic children in Libya. This retrospective study reviews our results as a developing country, comparing them with those in the literature, and makes suggestions to improve our results. Between May 1992 and May 1998, 94 children (58 males and 36 females) underwent 100 MBTSs in Mesallata Cardiothoracic Centre, Libya. The age ranged from 4 days to 15 years, (median 12 months) and 25 patients were neonates. Patients' weights ranged from 3 to 31 kg (median 6.4 kg). Eighty nine shunts were performed on the left side and 11 on the right. A 6-mm polytetrafluoroethylene graft was used in 68 children and a 4-mm graft in 32 cases. Tetralogy of Fallot (TOF) comprised the majority of cases (63; 67%), the remaining 31 (33%) included tricuspid atresia (13), pulmonary atresia (9), univentricular heart complex (6), and others (3). Acute shunt failure occurred in 3 cases (2 with 4-mm and 1 with 6-mm grafts), all of which had not received perioperative heparin. Hospital mortality was 6% (6 patients); risk factors were neonates, a diagnosis other than TOF, and emergency surgery. There were 12 late deaths, 6 of which were due to sepsis. Follow-up was achieved in 82 of 88 early survivors for a period of 2 to 60 months (median 26 months). All surviving patients had subjective and objective improvement. We conclude that MBTS is an excellent palliative procedure for children requiring a systemic-pulmonary shunt in developing as well as developed countries. This series shows an early mortality rate comparable to that of other studies, but the late mortality is higher, mainly due to sepsis, which warrants further attention. PMID- 11922509 TI - Experience with a DICOM-compatible digital pediatric echocardiography laboratory. AB - A digital pediatric echocardiography laboratory, without videotape redundancy was established at Children's Hospital of Wisconsin in December 1998. To characterize the experience, 1198 consecutive patient studies were reviewed-50% from the first 2 months after establishing the digital protocol and 50% from the last 2 months available. Each study was stored using a protocol that was based on capture of single beat clips of relevant two-dimensional/color Doppler imaging and static frame spectral Doppler tracings. Studies were digitally compressed using a DICOM compatible JPEG algorithm at 20:1 and edited with deletions of redundant clips to minimize archival storage needs. Study quality was uniformly excellent, and no errors were attributable to the digital protocol or compression-related loss of information. The average study required 21.5 +/- 11.4 MB of storage space with 35.4 +/- 12.3 total clips/study captured. Studies reviewed from the earlier experience were not significantly larger (23.6 +/- 14 vs 19.7 +/- 8.1 MB, 35.6 +/ 12.5 vs 35.2 +/- 12 clips) than those done recently. Studies in patients with isolated ventricular septal defect used comparable storage (23.7 +/- 8.9 MB, 42.8 +/- 11.5 clips) to that of the group as a whole. More complex congenital heart disease studies were slightly larger-tetralogy of Fallot (28.2 +/- 19.5 MB, 43.4 +/- 13.9 clips), transposition of the great arteries (30.6 +/- 17.4 MB, 40.3 +/- 16.7 clips), and single ventricle (29.7 +/- 19.6 MB, 39.9 +/- 12 clips)--although this trend was not significant. This study suggests that digital pediatric echo is feasible using a DICOM-compatible protocol with maintenance of diagnostic integrity despite compression of study size to allow rapid archival storage and retrieval. PMID- 11922510 TI - Noninvasive measurement of cardiac output in critically ill children. AB - This study was performed to evaluate the hemodynamic status of children admitted to the intensive care unit, using suprasternal and transesophageal Doppler ultrasound, and to establish a suitable noninvasive technique to monitor trends in cardiac output in critically ill children. Twenty children were studied over a period of 6 months. The median age was 32.5 months and weight 14.5 kg. Minute distance (MD), which is a linear cardiac output parameter, was assessed. Seven simultaneous pairs of measurements of MD were made using transesophageal Doppler (TED) and suprasternal Doppler (SSD) by the same operator. Following a fluid challenge, seven repeat pairs of measurements were made. The mean percentage changes for MD by TED and SSD were 21.84 (SD 9.97) and 5.75 (SD 7.32). The average coefficients of variation for measurements of MD by TED and SSD were 2.34% and 15.98%, respectively. The mean difference in percentage change between MD, measured by TED and SSD, was 27.59 with a 95% confidence interval and wide limits of agreement. The repeatability of TED measurements was good, but the measurements by SSD were wide and erratic with poor reproducibility. Our study shows that TED is easy to use, reliable, and very useful for monitoring hemodynamic changes in critically ill children. PMID- 11922511 TI - Coronary artery bypass grafting for Kawasaki disease. AB - Five patients with a history of Kawasaki disease underwent coronary revascularization at Children's Memorial Hospital (1988-2000). Acute disease occurred at 11 weeks to 5 years of age and revascularization procedures were performed at 8 months to 12 years (mean 6 years; interval from disease onset 5 months to 9 years). Surgical indications included abnormal stress testing with angiographic confirmation of severe coronary artery stenosis (n = 3), severe coronary artery stenosis with echocardiographic evidence of intracoronary thrombus (n = 1), and ischemic electrocardiogram changes and ventricular tachycardia during angiography (n = 1). All revascularization procedures used internal thoracic arteries including one free internal thoracic artery graft. There were no postoperative deaths (follow-up 1 month to 11 years). All patients are asymptomatic. One patient developed myocardial ischemia 4 years postoperatively with occlusion of the circumflex coronary artery (not previously grafted). This was treated successfully with percutaneous coronary angioplasty and stent placement. All grafts are patent with the exception of a single right internal thoracic artery graft which underwent involution 30 months postprocedure with concurrent recannulization of the right coronary artery. Coronary revascularization should be considered in the young patient with severe coronary abnormalities secondary to Kawasaki disease. PMID- 11922512 TI - Divided right atrium associated with extensive coronary vein abnormalities. AB - This report describes a case of a divided right atrium associated with coronary vein abnormalities. An 18-year-old woman who had a past history of surgery for repair of a divided right atrium and atrial septal defect developed exertional dyspnea 8 years after the surgery. Selective coronary angiography showed a dilated right coronary artery with a fistulous communication to the right atrium, tortuous coronary veins draining directly into the cardiac chambers, and the absence of the coronary sinus. Embryologically, regression failure of the right venous valve is hypothesized to have created both the divided right atrium and extensive coronary vein abnormalities. PMID- 11922514 TI - Successful treatment of a child with inferior vena cava thrombosis using a temporary inferior vena cava filter. AB - We describe a 6-year-old girl with Epstein-Barr virus-related hemophagocytic lymphohistiocytosis who developed inferior vena cava (IVC) thrombosis during chemotherapy. Treatment with an anticoagulant, thrombolytic agent and short-term placement of an IVC filter was followed by complete resolution. Pulmonary perfusion scintigraphy performed subsequently revealed no evidence of pulmonary embolism. We conclude that temporary placement of an IVC filter can be an important part of the treatment of IVC thrombosis in children. PMID- 11922513 TI - Cerebral and coronary embolization of a valvular tumor. AB - Stroke and myocardial infarction are not commonly seen in children; the occurrence of both conditions in the same well child is unusual. Undifferentiated cardiac tumors are rarely encountered in the pediatric population. We present a case of stroke and myocardial infarction as a consequence of an undifferentiated valvular tumor in a previously healthy child. This case emphasizes the critical importance of serial clinical examinations as part of a complete stroke workup. It suggests the need for additional research on the use of thrombolytic therapy for pediatric cerebral and myocardial infarction and stresses individualization of cardiac tumor treatment plans. PMID- 11922515 TI - Successful thrombolytic therapy of pulmonary embolism associated with urosepsis in an infant. AB - Childhood pulmonary embolism (PE) is a rare but serious condition marked by hypoxemia, shock, right-sided heart failure, and significant risk for fatality. Recommended treatment options include surgical embolectomy, anticoagulation, and thrombolysis. This report describes the successful use of recombinant tissue plasminogen activator to treat PE associated with urosepsis in a 34-day-old infant and reverse severe hemodynamic compromise. Diagnosis of proximal PE and monitoring its treatment were successfully achieved by echocardiography. PMID- 11922516 TI - Around PediHeart: aneurysm of the right ventricular apex in a 24-week fetus. PMID- 11922517 TI - Onset of complete atrioventricular block 15 years after ventricular septal defect surgery. AB - A 4-month-old girl with a ventricular septal defect underwent cardiac surgery. During the operation, complete atrioventricular block (CAVB) occurred, but on postoperative day 16 it converted to normal sinus rhythm with persistent complete right bundle branch block and left anterior hemiblock. Fourteen years after the operation, second-degree atrioventricular block was detected, but her general condition was good. Fifteen years after the operation, CAVB occurred. The intracardiac electrophysiologic recording revealed A-H and H-V block. A permanent pacemaker was implanted, and following the implantation she completely recovered. PMID- 11922518 TI - Around PediHeart: Amplatzer Septal Occluder device. PMID- 11922519 TI - Restrictive interatrial communication with protein-losing enteropathy and coagulopathy in hypoplastic left heart syndrome after Norwood palliation. AB - The Norwood procedure is one option for neonates born with hypoplastic left heart syndrome. We describe a case of an infant with hypoplastic left heart syndrome, palliatively repaired with the Norwood procedure. The infant developed restriction of the interatrial communication, despite atrioseptectomy at the first stage of palliation. Consequently, a protein-losing enteropathy with severe coagulopathy developed which resolved after a repeat atrioseptectomy. PMID- 11922520 TI - Accessory mitral valve tissue in combination with subaortic membrane and significant impairment of left ventricular function. AB - A 6-year-old girl was found to have a combination of accessory mitral valve tissue and subaortic membrane. The association caused left ventricular outflow tract obstruction leading to severe left ventricular dysfunction. Surgical relief of the obstruction resulted in immediate improvement of ventricular function. PMID- 11922521 TI - Coronary artery dilatation exceeding 4.0 mm during acute Kawasaki disease predicts a high probability of subsequent late intima-medial thickening. AB - We used intravascular ultrasound (IVUS) to compare the degree of coronary artery dilatation during the acute phase of Kawasaki disease with the extent of intima medial thickening more than 10 years later. We wanted to determine if there was a threshold degree of dilatation that was highly predictive of later thickening. Twenty-eight patients with a mean age of 17.3 +/- 1.7 years were studied; the mean interval from the initial selective coronary angiography to the IVUS study was 15.0 +/- 1.6 years. We measured the maximum intima medial thickness of selected coronary arterial segments in IVUS images and measured the largest diameters of the corresponding coronary arterial segments in the initial coronary angiograms. A significant correlation was found between the initial diameters of the coronary arteries and the intima medial thickness more than 10 years later in the right coronary, the left anterior descending coronary, and the left circumflex arteries. The coefficient of correlation was 0.77 (n = 120, p < 0.0001), and for the bifurcation of the left coronary artery it was 0.50 (n = 26, p < 0.01). For this study, abnormal intima medial thickness was defined as more than 0.40 mm. When the initial coronary arterial dilatation exceeded 4.0 mm, the sensitivity was 28/31 (90%) and the specificity was 87/89 (98%) in the right coronary, the left anterior descending coronary, and the left circumflex arteries. For the bifurcation of the left coronary artery, the sensitivity was 14/21 (67%) and the specificity was 5/5 (100%). PMID- 11922522 TI - Successful radiofrequency ablation of atypical left ventricular outflow tachycardia guided by epicardial activation signals. AB - A 12-year-old girl, with incessant atypical idiopathic left ventricular tachycardia, underwent successful radiofrequency ablation of ventricular tachycardia focus originating from a subepicardial site adjacent to the left coronary cusp. Ventricular tachycardia was successfully eliminated by targeting an endocardial site concordant to the epicardial site of early activation. Epicardial mapping was useful in locating an effective ablation site, and aortic root angiography was used to avoid potential injury to important structures, such as the left main coronary artery and left coronary cusp. PMID- 11922523 TI - Combining an MPH and/or a preventive medicine residency with family practice training. PMID- 11922525 TI - Doctors, patients ... and (excuse me!) interruptions. PMID- 11922526 TI - FP training expanded in People's Republic of China. PMID- 11922527 TI - Advising from a preceptor's perspective. PMID- 11922528 TI - Family medicine month in a university-based program: establishing a family medicine identity. AB - "Family Medicine Month," a rotation forfirst-year residents, was developed to clarifyfirst-year residents'new roles as family physicians. The rotation explored the meaning and history of the specialty, as well as teaching core family medicine clinical and behavioral skills. Twenty residents who participated in the rotation in 1999 and 2000 indicated satisfaction with the rotation and endorsed its usefulness. They also reported greater self efficacy in performing family practice skills after the rotation, compared with before the rotation. Results suggest that a curriculum based on the context of family medicine as a specialty enhances resident satisfaction and self efficacy. PMID- 11922529 TI - Family practice during a family practice residency. PMID- 11922530 TI - Malpractice: a poetic encounter. PMID- 11922531 TI - Resource-based relative value units: a primer for academic family physicians. AB - The Resource-based Relative Value Scale (RBRVS) is the prevailing model used to reimburse physician services today. Based on empirical research, relative value units (RVUs) quantify the relative work, practice expense, and malpractice costs for specific physician services to appropriately establish payment. The fee schedule, implemented by the Health Care Financing Administration in 1992, dramatically affected physician reimbursement, with the goal of correcting disparities across disciplines. In the first 6 years, Medicare payments to family physicians increased by 36%, while payments to specialists decreased by as much as 18%. Recent changes include new practice expense estimates and adjustment of payment based on facility type. The impact of RVUs is even more widespread as many private payers use the fee schedule to set payment rates and as RVUs become the yardstick for physician productivity measures. Despite the initial successes, the ability of RVUs to capture the work done by primary care providers is still limited. Primary care services today are not as easily quantified as surgical procedures, and coding limitations hinder documentation of services. Rapid changes in health care make comparisons to work done 2 decades ago difficult. Understanding the strengths and limitations of RVUs as they apply to family physicians is fundamental to safeguarding the role of primary care. PMID- 11922532 TI - The state of community medicine training in family practice residency programs. AB - OBJECTIVES: This paper describes the state and nature of community medicine training in family practice residency programs. METHODS: A random sample of 224 family practice residency programs was surveyed about the perceived value of community medicine in their residency, the teaching modalities they use, the extent to which their training provides competency in four defined dimensions of community medicine, and which program characteristics and curricula were predictive of higher perceived competency. RESULTS: The participation rate of our survey was 72%. Respondents ranked professional interest, institutional support, and departmental support highly. Less than half the programs provide instruction in community-oriented primary care (COPC), and less than half rate their department's involvement in the community highly. Most programs report that their training provides at least a moderate level of competency in four defined dimensions of community medicine. Curricular methods that are predictive of perceived competency include health department clinical experiences, home visits, cultural sensitivity training, participation in a longitudinal project, meetings with community leaders, and instruction in COPC. CONCLUSIONS: Community medicine is valued in residency curricula, but there is limited uniformity in curricular content and methods. Active and structured education modalities might be more likely to result in competency in community medicine. PMID- 11922533 TI - Prevalence of community-oriented primary care knowledge, training, and practice. AB - BACKGROUND AND OBJECTIVES: Recent recommendations requiring resident training in community-oriented primary care (COPC) indicate a continued interest among family medicine educators. This study examines COPC-related aspects of training and practice and whether or not respondents report COPC knowledge. The study also compares residency program and physician responses. METHODS: A total of 400 randomly selected practicing physicians and 470 residency directors were asked about COPC curricular and practice experiences. Physicians were asked if they practice COPC. Programs were asked if they taught COPC. Both were asked if they were knowledgeable about COPC. RESULTS: Response rates for practicing physicians and programs were 58.4% and 71.8%, respectively; 38.8% of programs teach COPC, and 6.7% of physicians reported that they practice COPC. Sixty-seven percent of programs and 19% of physicians reported COPC knowledge. Programs with knowledge of COPC conducted more COPC-related activities than those without such knowledge. This relationship was not seen among practicing physicians. CONCLUSIONS: Aspects of COPC exist in training and in practice environments. Knowledge about COPC is associated with differences in programs' COPC activities but not in the COPC activities of practicing physicians. Programs and physicians differ in COPC implementation in training and practice. PMID- 11922534 TI - Working with impaired residents: trials, tribulations, and successes. AB - Impairment of physicians' ability to practice medicine safely and effectively is relatively common. Chemical dependency, the leading cause of physician impairment, has a lifetime prevalence of approximately 10%-15% among physicians. Statistics from physician health programs indicate that family physicians are overrepresented among impaired physicians. It is therefore important for family practice residencies to monitor for and deal with physician impairment. Over the past 11 years, we have worked with eight impaired residents: five with chemical dependency, two with cognitive impairment, and one with an affective disorder Seven of the eight residents are currently practicing medicine, six in family practice. Based on our experience and the literature, we have developed an algorithm that includes the recognition, intervention, and aftercare of impaired residents. The long-term success of the majority of impaired residents with whom we have worked suggests that the trials and tribulations of working with this potentially difficult group of residents are well worth the effort. PMID- 11922535 TI - Self-directed learning: looking at outcomes with medical students. AB - BACKGROUND AND OBJECTIVES: Self-directed learning (SDL) skills are thought to be associated with lifelong learning. This study assessed the degree of readiness for SDL in third-year medical students who participated in a problem-based learning (PBL) curriculum during thefirst 2 years of medical school. METHODS: A total of 182 third-year medical students at the University of Texas Medical Branch at Galveston were given the Self-directed Learning Readiness Scale (SDLRS). RESULTS: The observed mean (235.81 [range 183-284]) for the combined group was significantly higher than the mean reported for general adult learners (214), though slightly lower than scores reported in studies of other medical students and professionals. Ratings of students by clinical preceptors correlated with SDLRS scores. CONCLUSIONS: Students in our integrated medical curriculum had scores on the SDLRS that correlated with clinical performance and probably represented a readiness for SDL. PMID- 11922536 TI - Use of a peripheral dexa measurement for osteoporosis screening. AB - BACKGROUND AND OBJECTIVES: Osteoporosis is a substantial cause of morbidity and mortality in the United States. While screening for this disease is important, few studies have evaluated the role of an osteoporosis screening device in afamily practice setting. This study evaluated the influence of a peripheral-dexa (p-dexa) heel bone mineral density (BMD) measurement and a patient education program on changes in pharmacologic treatment for suspected osteopenia or osteoporosis by primary care physicians over a 1-year follow-up. METHODS: Using a computerized database (ages 50 to 75 years), 1,927 women were identified. An invitation was mailed to register for a screening to have a p-dexa heel BMD scan done and to attend an osteoporosis prevention presentation. Medication history, age, height, weight, and risk factors for osteoporosis were collected. A follow up chart review was carried out on women who werefound to have heel BMD T-scores of <-.6 (suggested by the World Health Organization). Date of menopause onset, pharmacotherapy for osteoporosis, calcium, vitamin D, and physician intervention were assessed. RESULTS: There were 292 women (15.2%) who self registered, obtained BMD testing, and attended an educational program. Of these women, 87 (30%) had at least one risk factor for osteoporosis, in addition to menopause. Mean BMD was .489 +/- .113 gm/cm2 (normal >.42 gm/cm2). A post-screening chart review was completed in 102 women (36.6%) at greatest risk for osteoporosis based on a T-score <-.6. Following the intervention, 26 women were started on antiresorptive therapy (primarily estrogen), and three additional women had a second antiresorptive agent added to estrogen. CONCLUSIONS: P-dexa heel BMD has utilityfor screening patients at riskfor osteoporosis. However, only 15% of invited women attended the screenings, and pharmacotherapy treatment did not significantly change after screening in the majority of women at riskfor osteoporosis, based on p-dexa screening. PMID- 11922537 TI - Backlash [editorial]. PMID- 11922538 TI - Reflections from the cave: an essay on video precepting. PMID- 11922539 TI - Inadequate follow-up time bias: the appearance of genetic anticipation and temporal trends in IBD. PMID- 11922540 TI - Cancer family history and genetic testing: are malpractice adjudications waiting to happen? PMID- 11922541 TI - The natural history of HCV infection in African Americans. PMID- 11922542 TI - How many times have you done this procedure, doctor? PMID- 11922543 TI - Management of primary sclerosing cholangitis. AB - Guidelines for clinical practice are intended to indicate preferred approaches to medical problems as established by scientifically valid research. Double blind, placebo-controlled studies are preferable, but reports and expert review articles are also utilized in a thorough review of the literature conducted through the National Library of Medicine's MEDLINE. When only data that will not withstand objective scrutiny are available, a recommendation is identified as a consensus of experts. Guidelines are applicable to all physicians who address the subject, without regard to specialty training or interests, and are intended to indicate the preferable but not necessarily the only acceptable approach to a specific problem. Guidelines are intended to be flexible and must be distinguished from standards of care that are inflexible and rarely violated. Given the wide range of specifics in any health care problem, the physician must always choose the course best suited to the individual patient and the variables in existence at the moment of decision. Guidelines are developed under the auspices of the American College of Gastroenterology and its Practice Parameters Committee and approved by the Board of Trustees. Each has been intensely reviewed and revised by the Committee, other experts in the field, physicians who will use them, and specialists in the science of decision of analysis. The recommendations of each guideline are therefore considered valid at the time of their production based on the data available. New developments in medical research and practice pertinent to each guideline will be reviewed at an established time and indicated at publication to assure continued validity. PMID- 11922544 TI - Portal vein thrombosis: a concise review. AB - Portal vein thrombosis (PVT) is an uncommon cause for presinusoidal portal hypertension. Although several predisposing conditions are known to exist in the background of PVT, there still remains a proportion of patients in whom the etiology is not known and the pathogenesis is unclear. In this review we summarize the literature on PVT and present the current knowledge about the precipitating factors of PVT. Further, we discuss the advances in the radiological diagnosis that have improved diagnostic accuracy and are noninvasive. Finally, we discuss the treatment options for patients who have varying extents of thrombosis in the portal vein and specifically focus on PVT that is encountered before and after liver transplantation. PMID- 11922545 TI - NSAID inhibition of GI cancer growth: clinical implications and molecular mechanisms of action. AB - Epidemiological studies suggest that aspirin and other non-steroidal anti inflammatory drugs (NSAIDs) reduce the incidence of and mortality from colorectal, gastric, and esophageal cancers. The precise mechanisms by which NSAIDs exert their chemopreventive effects are not fully explained, but likely involve inhibition of cyclo-oxygenase, the enzyme that converts arachidonic acid to prostaglandins. Two isoforms of this enzyme, cyclo-oxygenase 1 (COX-1) and COX 2, have been identified. COX-2 is absent in normal mucosa but is overexpressed in colonic, gastric, and esophageal cancers, as well as their precursor lesions. The inhibition of COX-2 through either pharmacological agents or gene deletion results in suppression of colonic polyp formation. NSAIDs reduce colonic, gastric, and esophageal cancer cell growth, in part, by inducing apoptosis. However, the antineoplastic effects of NSAIDs may be partly independent of their ability to inhibit COX-2. The mechanisms involved in the antineoplastic actions of NSAIDs include inhibition of angiogenesis (essential for delivery of oxygen and nutrients to a growing tumor), induction of apoptosis (which is usually reduced in cancer cells) by stimulation of proapoptotic genes, and direct inhibition of cancer cell growth by blocking signal transduction pathways responsible for cell proliferation. PMID- 11922546 TI - Clinical, endoscopic, and functional studies in 408 patients with Barrett's esophagus, compared to 174 cases of intestinal metaplasia of the cardia. AB - OBJECTIVE: The pathophysiology of gastroesophageal reflux disease (GERD) has been studied extensively in patients with long-segment Barrett's esophagus (LSBE), but few reports have explored GERD pathophysiology in patients who have short-segment Barrett's esophagus (SSBE) or intestinal metaplasia at the cardia (IMC). We aimed to compare clinical, endoscopic, histological, and functional features in patients with LSBE, SSBE, and IMC. METHODS: We identified 582 patients who had intestinal metaplasia at the squamocolumnar junction in the distal esophagus and divided them into three groups based on the extent of columnar-lined esophagus observed endoscopically: 1) patients with IMC who had no columnar-lined esophagus (i.e., the squamocolumnar and gastroesophageal junctions coincided), 2) patients with LSBE who had >3 cm of columnar-lined esophagus, and 3) patients with SSBE who had <3 cm of columnar-lined esophagus. All patients had esophageal manometric evaluation, and 24-h esophageal pH monitoring was performed to determine the extent of acid and bile (bilirubin) reflux. RESULTS: There were 174 patients with IMC, 155 with LSBE, and 25 with SSBE. Compared to patients with LSBE and SSBE, patients with IMC had significantly lower frequencies of GERD symptoms, hiatal hernia, and erosive esophagitis; significantly higher lower esophageal sphincter pressures; and significantly shorter durations of acid and bile reflux. Between patients with SSBE and LSBE, significant differences were found in the frequency of hiatal hernia and duration of acid reflux (both greater in the patients with LSBE). Also, dysplasia was significantly more frequent in patients with LSBE than in those with SSBE or IMC. CONCLUSION: GERD symptoms, signs, and physiological abnormalities are found more often in patients with Barrett's esophagus than in those with IMC, and the duration of acid reflux in patients with LSBE is greater than that in patients with SSBE. These findings suggest that the extent of intestinal metaplasia in the esophagus is related directly to the severity of underlying GERD. PMID- 11922547 TI - Gastroesophageal reflux induced by white wine: the role of acid clearance and "rereflux". AB - OBJECTIVES: White wine has been demonstrated to induce gastroesophageal reflux (GER) in healthy people and GER patients. This GER is characterized by reflux episodes of prolonged duration. Our aim was to explore the pathogenesis of the prolonged reflux duration. METHODS: Twelve healthy volunteers received in a randomized order 300 ml of white wine and tap water together with a standardized meal. Esophageal pH and motility were continuously monitored by a glass pH electrode and a strain gauge manometry probe (four measuring points in the esophagus and one in the pharynx to register swallowing) for 90 min after ingestion. Blinded to the ingested beverage, we calculated the fraction of time esophageal pH was <4, the number of reflux episodes and their duration, the swallowing and contraction rate, the contraction amplitude, and the distribution of primary, secondary, simultaneous, and nonpropagated contractions. The motility analysis was separately performed for periods with and without GER. During GER, the time until occurrence of the first contraction, its type, the type of the contraction that raises pH to >4, and the number of peristaltic contractions necessary to raise pH to >4 were also determined. The percentage of GER episodes with simultaneous contractions and failed peristalsis (nontransmitted swallows and nonpropagated contractions) as calculated. The percentage of GER episodes with signs of "rereflux" (further pH drop, common cavity phenomenon in the motility trace) into the acidic esophagus was also determined. The mean reflux duration and the number of peristaltic contractions needed to raise pH to >4 were recalculated by taking the rereflux events into account. RESULTS: White wine significantly increased the fraction of time esophageal pH was <4, reflux frequency, and reflux duration compared to water. During periods without GER, no differences in the motility data were observed between wine and water. During GER, the contraction rate after white wine was significantly lower because of an increase in nontransmitted swallows. The time until occurrence of the first contraction after GER was significantly prolonged after white wine. Primary peristalsis was the main first and clearance contraction type. The percentage of GER episodes with simultaneous contractions and with failed peristalsis was significantly increased with wine. Similarly, the percentage of GER episodes with rereflux was significantly increased. The "corrected" mean reflux duration was still prolonged relative to water, but the difference was no longer significant. The numbers of peristaltic contractions necessary to raise pH to >4, which significantly differed for conventionally defined GER episodes between wine and water, were similar when counted only from onset of the latest rereflux event until pH rose to >4. CONCLUSIONS: The pathogenesis of white wine-induced GER episodes of long duration is 2-fold. First, white wine provokes a disturbed esophageal clearance due to an increase in simultaneous contractions and in failed peristalsis. The second mechanism is the occurrence of repeated reflux events into the esophagus when pH is still acidic from a previous reflux episode. PMID- 11922548 TI - Improving the analysis of esophageal acid exposure by a new parameter: area under H+. AB - OBJECTIVES: We aimed to compare the data provided by 24-h continuous esophageal pH monitoring in a group of patients with gastroesophageal reflux disease (GERD) to those from a group of healthy volunteers using both conventional parameters and calculated area under the curve of hydrogen ion activity (AUH+), a new value that describes the true acid exposure, through both duration and depth of acidity changes. METHODS: Thirty healthy controls and 60 patients with GERD (30 symptomatic patients without endoscopic esophagitis or nonerosive GERD and 30 symptomatic patients with Savary I-IV endoscopic esophagitis or erosive GERD) were enrolled in a study based on 24-h pH monitoring to compare reference values by means of receiver operating characteristic (ROC) discriminant analysis. RESULTS: The best ROC cutoff value for nonerosive GERD patients was AUH+ = 103.7 (mmol/L) x min with sensitivity of 76.7% and specificity of 93.3%. The best ROC cutoff value for erosive GERD patients was AUH+ = 114.1 (mmol/L) x min with sensitivity of 100% and specificity of 96.7%. These cutoff values increase the sensitivity by 16.7% for nonerosive GERD patients and 10% for erosive GERD patients when compared to a common parameter such as the percentage of total time pH is <4 with a limit of 4.2%. CONCLUSIONS: AUH+ is a valid quantitative parameter to measure 24-h esophageal acid exposure. It may be a reliable and significant clinical aid because it is a more sensitive test in discriminating negative or positive adult patients with or without esophagitis who are submitted to 24-h esophageal pH monitoring. PMID- 11922550 TI - Intestinal metaplasia of the gastric cardia: A prospective study with enhanced magnification endoscopy. AB - OBJECTIVE: There are no endoscopic features that distinguish intestinal metaplasia of the cardia (CIM) from the normal cardia. Biopsy specimens are therefore randomly obtained from normal-appearing mucosa with significant potential sampling errors. Enhanced magnification endoscopy involves the combined use of magnification endoscopy with acetic acid instillation. This study assessed the value of enhanced magnification endoscopy in detecting CIM. METHODS: Patients undergoing elective upper endoscopy were invited to participate in the study. Patients were included if the squamocolumnar junction and the esophagogastric junction were judged to be at the same level. Enhanced magnification endoscopy was performed with 3% acetic acid instillation. Standard endoscopy was followed by magnification endoscopy and repeated after acetic acid spraying. Surface patterns were characterized before and after acetic acid spraying. The observed surface patterns were compared with histological results obtained from a single targeted biopsy specimen of each pattern. RESULTS: The overall prevalence of CIM was 34.8% (86/247 patients). After excluding 52 patients because of endoscopic evidence of Barrett's esophagus, 195 patients were eligible for participation in the study. In the study group, CIM was detected in 86 patients (44.1%) in targeted biopsy samples. No dysplasia was identified. Enhanced magnification endoscopy detected four different patterns of the mucosal surface: I) round pits, II) reticular, III) villous, and IV) ridged. The yields of detection of intestinal metaplasia according to endoscopic patterns were I) 0%, II) 5.3% (odds ratio = 0.05), III) 57.7% (odds ratio = 7.5, p = 0.0001), and IV) 95.8% (odds ratio = 42.8, p = 0.0001). CONCLUSIONS: CIM is more common than previously reported. Enhanced magnification endoscopy identifies two characteristic endoscopic patterns, villous (pattern III) and ridged (pattern IV), with outstanding clarity and resolution that correlate with histological identification of CIM with a single targeted biopsy sample. Enhanced magnification endoscopy will permit longitudinal studies of an entity that can be identified endoscopically. PMID- 11922549 TI - Esomeprazole (40 mg) compared with lansoprazole (30 mg) in the treatment of erosive esophagitis. AB - OBJECTIVES: Esomeprazole, the S isomer of omeprazole, has been shown to have higher healing rates of erosive esophagitis than omeprazole. This study compared esomeprazole with lansoprazole for the healing of erosive esophagitis and resolution of heartburn. METHODS: This United States multicenter, randomized, double blind, parallel group trial was performed in 5241 adult patients (intent to-treat population) with endoscopically documented erosive esophagitis, which was graded by severity at baseline (Los Angeles classification). Patients received 40 mg of esomeprazole (n = 2624) or 30 mg of lansoprazole (n = 2617) once daily before breakfast for up to 8 wk. The primary efficacy endpoint was healing of erosive esophagitis at week 8. Secondary assessments included proportion of patients healed at week 4, resolution of investigator-recorded heartburn, time to first and time to sustained resolution of patient diary recorded heartburn, and proportion of heartburn-free days and nights. RESULTS: Esomeprazole (40 mg) demonstrated significantly higher healing rates (92.6%, 95% CI = 91.5-93.6%) than lansoprazole (30 mg) (88.8%, 95% CI = 87.5-90.0%) at week 8 (p = 0.0001, life-table estimates, intent-to-treat analysis). A significant difference in healing rates favoring esomeprazole was also observed at week 4. The difference in healing rates between esomeprazole and lansoprazole increased as baseline severity of erosive esophagitis increased. Sustained resolution of heartburn occurred faster and in more patients treated with esomeprazole. Sustained resolution of nocturnal heartburn also occurred faster with esomeprazole. Both treatments were well tolerated. CONCLUSIONS: Esomeprazole (40 mg) is more effective than lansoprazole (30 mg) in healing erosive esophagitis and resolving heartburn. Healing rates are consistently high with esomeprazole, irrespective of baseline disease severity. PMID- 11922551 TI - Predictive factors of GI lesions in 241 women with iron deficiency anemia. AB - OBJECTIVES: GI blood loss is the most common cause of iron deficiency anemia (IDA) in postmenopausal women and menstrual blood loss in premenopausal women. We aimed to evaluate the diagnostic yield of endoscopy in women with IDA and to define predictive factors of a GI lesion. METHOD: Clinical, biological, endoscopic, and histological data from patients with IDA were systematically collected on a computer. Multivariate analysis (logistic regression) was performed to determine whether these data were associated with a GI lesion. RESULTS: Between January, 1989 and June, 1999, 241 consecutive women had endoscopies for IDA (mean age = 52.3 +/- 21.8 yr). A substantial GI lesion was detected in 119 patients (49.4%). Ten patients (4%) had both upper and lower GI lesions. A source of IDA was revealed by upper endoscopy in 86 cases (35.6%) and by colonoscopy in 33 (13.7%). The most common upper lesions were peptic ulceration (42/241 [17.4%]), esophagitis (15/241 [6.2%]), and cancer (9/241 [3.7%]). Colonic cancer (15/241 [6.2%]) and polyps (10/241 [4.1%]) were the most frequent lesions detected by colonoscopy. Predictive factors (odds ratio, 95% CI) of GI lesions diagnosed by endoscopy were abdominal symptoms (8.3, 3.9-17.2), age > 50 yr (4.4, 2.1-9.2), and Hb < 9 g/dl (3, 1.5-6.1). Thirty-one women (13%) had none of these predictive factors; in this group only two lesions were identified (one esophagitis and one duodenal ulcer). The positive predictive value of these three independent predictors was 87%, and the negative predictive value was 93.5%. CONCLUSION: Endoscopy revealed a source of IDA in 49.4% of cases. Three predictive factors of GI lesion were identified. Endoscopic investigation should be avoided in women without these three predictive factors. Conversely, these factors are strongly associated with a GI lesion. PMID- 11922552 TI - Evaluation of the Helicobacter pylori stool antigen test for monitoring eradication therapy. AB - OBJECTIVE: We aimed to clarify the time course of the Helicobacter pylori stool antigen (HpSA) level during and after eradication therapy and to determine the appropriate time of measurement of HpSA to evaluate eradication. METHODS: The subjects were 47 patients who were positive for H. pylori. The patients underwent a 1-wk regimen of triple therapy. The outcome of the eradication therapy was judged by urea breath test, culture, and histology 6 wk after the end of treatment. The HpSA level was serially measured nine times from before therapy until 12 wk after the end of therapy. RESULTS: In the group with successful eradication, HpSA became negative immediately after the end of therapy and the negativity persisted. In contrast, in the noneradication group HpSA became negative immediately after therapy, but became positive again within 2 wk after the end of therapy. The mean absorbance value of the HpSA test on the 4th day after the initiation of eradication therapy was significantly higher in the noneradication group than in the group with successful eradication. The diagnostic accuracy of the HpSA test increased to > or = 90% 2 wk after therapy and thereafter. Comparison of the diagnostic accuracy at 4 wk after the end of therapy showed no significant differences with that at 2, 6, 8, and 12 wk after the end of therapy. CONCLUSIONS: The course of the HpSA levels during and after eradication therapy in patients with successful eradication was quite different from that in noneradication patients. Measurement of HpSA was useful to evaluate eradication, and the appropriate evaluation of the outcome of treatment could be made as early as 2 wk after the end of therapy. PMID- 11922553 TI - Upper GI malignancy, uncomplicated dyspepsia, and the age threshold for early endoscopy. AB - OBJECTIVES: We sought to determine the frequency with which patients with upper GI malignancies present with uncomplicated dyspepsia and to determine the impact of changing the threshold for early endoscopy from 45 to 55 yr, as has recently been recommended in European studies. METHODS: Three hundred forty-one patients with upper GI malignancies at two Milwaukee teaching hospitals were identified from electronic databases. Trained physician-investigators reviewed the charts using a specifically designed questionnaire. Staging of GI cancers was performed using the TNM classification to determine operability. Uncomplicated dyspepsia was defined as dyspepsia without alarm symptoms/signs (dysphagia, vomiting, weight loss, early satiety, GI bleeding, or anemia). RESULTS: There were 341 patients with upper GI malignancies with a mean age of 68 yr (range = 22-94). Twenty-one patients were younger than 45 and 65 were under 55. Uncomplicated dyspepsia was the presenting symptom in five of 65 (7.7%) patients younger than 55 and one of 21 (4.8%) patients under 45 (p = 0.99). Five patients 55 or younger presented with uncomplicated dyspepsia. All had advanced, inoperable disease (stage III or higher). CONCLUSIONS: 1) In a racially diverse United States population, upper GI malignancy presenting without alarm symptoms is uncommon. 2) Raising the threshold for immediate endoscopy in uncomplicated dyspepsia from age 45 to 55 increases the risk of missing an underlying malignancy but may not affect outcome, as incurable disease was present at diagnosis within a few weeks of the onset of symptoms. PMID- 11922554 TI - GI symptoms in diabetes mellitus are associated with both poor glycemic control and diabetic complications. AB - OBJECTIVE: Diabetes mellitus is associated with an increased prevalence of GI symptoms, but the mechanisms underlying symptoms are poorly defined and controversial. We aimed to determine whether there is a relationship between GI symptoms and both diabetic complications and glycemic control. METHODS: We performed a cross-sectional questionnaire study of 1101 subjects with diabetes mellitus recruited from outpatient clinics (n = 209) and the community (n = 892). Data on eight GI symptom groups, complications of diabetes (retinopathy, neuropathy, nephropathy), and self-reported glycemic control were obtained from a validated questionnaire. Glycated hemoglobin was measured in 463 of the subjects, The association between diabetic complications, glycemic control, and GI symptoms was assessed using logistic regression analysis, adjusted for demographic and clinical factors. RESULTS: Of the 1101 subjects, 57% reported at least one complication. Diabetic complications were independently associated with both symptom complexity (number of symptom groups reported) (adjusted odds ratio = 1.92 per symptom group [95% CI = 1.51-2.45]) and seven of the eight GI symptom groups. For all symptom groups, the association was explained by self-reported symptoms of peripheral neuropathy. Poor glycemic control measured by both self report and Hb A1c was an independent risk factor for upper GI symptoms, whereas other potential risk indicators, including duration and type of diabetes, were not significant. CONCLUSIONS: GI symptoms in diabetes mellitus may be linked to diabetic complications, particularly peripheral neuropathy, and to poor glycemic control. PMID- 11922555 TI - Octreotide relaxes the hypertensive sphincter of Oddi: pathophysiological and therapeutic implications. AB - OBJECTIVES: As our understanding of the pathophysiology of sphincter of Oddi dysfunction (SOD) expands, new avenues arise for pharmacological intervention. Recent evidence suggests that SOD results from a loss of myenteric plexus inhibitory neurons resulting in unopposed cholinergic tone. Octreotide inhibits postganglionic cholinergic neurons, and thus we hypothesize that administration of octreotide will decrease sphincteric pressure in individuals with SOD. METHODS: Thirty-eight patients presenting with recurrent abdominal pain and SOD (basal pressure > 40 mm Hg) were studied. The study was prospective, placebo controlled, and blinded. Patient allocation was consecutive. Sphincter of Oddi manometry was performed in standard fashion. The test group (n = 19) received octreotide acetate (100 microg i.v.), and the control group (n = 19) received i.v. saline. Basal, phasic, and duct pressures as well as phasic amplitude and frequency were recorded before and 3 min after the i.v. infusion. Changes in these parameters before and after i.v. infusions were compared. RESULTS: Octreotide caused a statistically significant reduction in peak and basal sphincter of Oddi pressures relative to saline (p < 0.01 and p < 0.001). Octreotide did not significantly affect wave amplitude, wave frequency, or duct pressure. CONCLUSIONS: Octreotide has the potential to be a valuable addition to the armamentarium for the medical management of SOD. PMID- 11922556 TI - Electronic pancreatoscopy for the diagnosis of pancreatic diseases. AB - OBJECTIVES: Diagnostic pancreatoscopy was performed in 56 patients using the newly developed peroral electronic pancreatoscope (PEPS). The diagnostic value of the PEPS for pancreatic diseases was evaluated. METHODS: The PEPS (external diameter = 2.1 mm, bidirectional tip deflection) was developed with a minute 50,000-pixel interline charge-coupled device. Pancreatoscopy was performed by means of an endoscopic retrograde approach without sphincterotomy. The cases examined were normal (three), chronic pancreatitis (32), pancreatic cancer (eight), and intraductal papillary mucinous tumors (13). RESULTS: Of the 56 cases, 42 (75%) were adequately seen. In normal cases, fine capillary vessels were clearly visualized on the smooth whitish-pink mucosa. Findings in chronic pancreatitis included protein plugs, calcified stones, rough whitish mucosa, scar formation, edema, erythema, and indistinct capillary appearance. All the stenoses of chronic pancreatitis could be differentiated from those of pancreatic cancer with the PEPS. In the pancreatic cancer cases, all patients had stenoses or duct cutoffs; most cases had friable mucosa with erythema and erosive changes, and a single patient had a compressed pancreatic duct wall covered with normal epithelium. In the cases of intraductal papillary mucinous tumors, papillary tumors were visualized with extreme clarity. In the case of adenocarcinoma, the PEPS revealed oval-shaped tumors with spotty redness or villous tumors with dilation of capillary vessels. Moderate acute pancreatitis was recognized after pancreatoscopy in one of the 56 cases (1.8%). CONCLUSIONS: Electronic pancreatoscopy with the PEPS is feasible in most patients and technically safe, and improves diagnostic yield over conventional pancreatoscopy. PMID- 11922557 TI - Combined ligation and sclerotherapy versus ligation alone for secondary prophylaxis of esophageal variceal bleeding: a meta-analysis. AB - OBJECTIVES: Variceal ligation has been shown to be superior to sclerotherapy in prevention of rebleeding and improving survival in patients with cirrhosis. However, 25% of patients will rebleed before completion of treatment. A number of trials have compared the combination of ligation and sclerotherapy to ligation alone in achieving rapid and complete eradication of esophageal varices, with conflicting results. METHODS: Two reviewers independently identified seven randomized, controlled trials that compared endoscopic variceal ligation with the combination of sclerotherapy and ligation for the treatment of esophageal varices. Studies were identified by searching MEDLINE, reviewing references from retrieved articles, and scanning abstracts from conference proceedings. For each outcome, odds ratios (ORs) were calculated using fixed-effects and random-effects models. The Mantel-Haenszel test for statistical heterogeneity was used to assess the validity of combining results from individual studies. RESULTS: No significant difference was seen in cessation of actively bleeding varices (OR = 1.01, 95% CI = 0.43-2.36), variceal rebleeding (OR = 1.12, CI = 0.69-1.81), and mortality (OR = 1.1, CI = 0.70-1.74) in patients with variceal ligation versus patients receiving the combination treatment of ligation and sclerotherapy. Treatment sessions required to achieve complete variceal eradication were similar in the two treatment arms. A significantly higher incidence of esophageal stricture was seen in combination therapy (p < 0.001). CONCLUSIONS: The combination of ligation and sclerotherapy offers no advantage over ligation alone in prevention of rebleeding and in reduction of mortality. It is also associated with a higher complication rate of esophageal stricture. PMID- 11922559 TI - Generational differences in the age at diagnosis with Ibd: genetic anticipation, bias, or temporal effects. AB - OBJECTIVE: Previous cross-sectional research has demonstrated generational differences in age at diagnosis (AAD) in inflammatory bowel disease (IBD). This observation has at times been ascribed to genetic anticipation, but could also be due to biases related to case ascertainment or follow-up or to temporal changes in IBD epidemiology. We aimed to explore this issue using a population-based database. METHODS: In 1995 we used the comprehensive administrative databases in the province of Manitoba, Canada to establish a population-based IBD Research Registry that includes clinical and demographic information for persons. We contacted those subjects within our Research Registry who reported having any family members with IBD and their family members for verification of diagnosis and AAD. Differences in AAD between familial pairs were calculated. In addition, to assess whether duration of follow-up accounted for generational differences in AAD, we computed the mean AAD for subjects with and without family histories of IBD based on age at the time of interview (i.e., < 45 and > or = 45 yr of age). RESULTS: Of the 2445 persons with IBD in the Research Registry, 548 reported positive family histories, and 315 of these (58%) were reached by telephone. There were 169 Crohn's disease and 146 ulcerative colitis subjects with positive family histories. The mean AADs for the parents, aunts/uncles, and grandparents were significantly greater than the mean AADs for the children, nieces/nephews, and grandchildren, respectively. There was a doubling of the mean AAD when comparing the grandparent/grandchild cases with the parent/child or aunt/uncle niece/nephew cases. No statistically significant difference in anticipation was observed, whether or not the older generation was male or female or had Crohn's disease or ulcerative colitis. The AAD was substantially greater for those interviewed at > or = 45 yr of age for subjects with and without family histories. However, there was no substantial difference in mean AAD between familial and nonfamilial subjects. CONCLUSION: The present study has demonstrated that there is a tendency for children to be younger than their parents at the time of diagnosis of familial IBD, and that this difference in mean AAD is almost doubled for grandparent/grandchild pairs. However, we conclude that these differences are most likely due to a bias based on length of follow-up or recent multigenerational temporal changes in the risk of IBD, or both. PMID- 11922558 TI - Prospective validation of the Rockall risk scoring system for upper GI hemorrhage in subgroups of patients with varices and peptic ulcers. AB - OBJECTIVES: The Rockall risk assessment score was devised to allow prediction of the risk of rebleeding and death in patients with upper GI hemorrhage. The score was derived by multivariate analysis in a cohort of patients with upper GI hemorrhage and subsequently validated in a second cohort. Only 4.4% of patients included in the initial study had esophageal varices, and analysis was not performed according to the etiology of the bleeding. Our aim was to assess the validity of the Rockall risk scoring system in predicting rebleeding and mortality in patients with esophageal varices or peptic ulcers. METHODS: Admissions (n = 358) over 32 months to a single specialist GI bleeding unit were scored prospectively. The distribution of episodes of rebleeding and mortality by Rockall score were statistically analyzed using Fisher's exact test with 99% CIs calculated using a Monte Carlo method. The Child-Pugh score was determined in patients with esophageal varices. RESULTS: The Rockall score was predictive of both rebleeding and mortality in patients with variceal hemorrhage (both ps < 0.0005), as was the Child-Pugh score (p = 0.001 and p < 0.0005, respectively). The initial Rockall score was predictive of mortality in patients with peptic ulcers (p = 0.01), although the complete score was not (p > 0.05). The complete score did, however, predict rebleeding in these patients (p = 0.001). CONCLUSION: This is the first study to validate the Rockall score in specific subgroups of patients with esophageal varices or peptic ulcers and suggests that it is particularly applicable to variceal hemorrhage. PMID- 11922560 TI - Budesonide and mesalazine in active Crohn's disease: a comparison of the effects on quality of life. AB - OBJECTIVES: Controlled ileal release budesonide and slow release mesalazine are both used to treat mild to moderate active Crohn's disease, although data show that budesonide is more effective in inducing remission. When comparing different treatment options, the effects of agents on health-related quality of life must be considered as well as efficacy. In this study, we sought to compare the effects of budesonide and mesalazine on the health-related quality of life of patients with active Crohn's disease. METHODS: The study included 182 patients with Crohn's Disease Activity Index scores between 200 and 400. Patients were randomized in a double blind, double dummy, multicenter study to receive 9 mg of budesonide, once daily (n = 93), or 2 g of mesalazine, b.i.d. (n = 89), for 16 wk. Quality of life was assessed at baseline and after 2, 4, 8, 12, and 16 wk of treatment using the Psychological General Well-Being index. In addition, a physician's global evaluation was used to assess how symptoms affected patients' normal activities. RESULTS: Patients treated with budesonide experienced significantly greater improvement in Psychological General Well-Being scores than the group treated with mesalazine after 2, 8, 12, and 16 wk. All components of this index showed greater improvements in the budesonide-treated group than in the mesalazine group at 12 and 16 wk. The physician's global evaluation showed significantly greater improvements in the budesonide group than in the mesalazine group at all visits. CONCLUSION: Budesonide (9 mg once daily) improves health related quality of life to a greater extent than mesalazine (2 g b.i.d.) in patients with mild to moderate active Crohn's disease. PMID- 11922561 TI - Outcome of patients with radiation enteritis treated with home parenteral nutrition. AB - OBJECTIVES: Intestinal failure requiring either surgery or home parenteral nutrition (HPN) develops in approximately 5% of patients treated with radiation. The aim of the study was to determine survival, duration of HPN, and complications associated with HPN in patients with intestinal failure after radiation therapy. METHODS: Fifty-four patients with radiation enteritis who received HPN were studied (39 women and 15 men with a mean age of 57.9 yr). Retrospective data were collected from the patients' medical records dated between 1975 and 1999. The probability of survival was calculated by the Kaplan Meier method. RESULTS: HPN was initiated a median of 20 months (range = 2-432) from the start of radiation therapy. The mean number of intestinal operations for radiation-related complications was 2.2/patient (range = 0-6). The causes of intestinal failure resulting from radiation therapy were intestinal obstruction (27 patients), short bowel (17), malabsorption (five), fistula (three), and dysmotility (two). The mean duration of HPN was 20.4 months (range = 2-108). At last follow-up, 37 patients (68%) were dead, most as a result of recurrent cancer. One patient died of catheter sepsis, and no other deaths were directly related to HPN. The overall estimated 5-yr probability of survival on HPN calculated by Kaplan-Meier analysis was 64%. CONCLUSIONS: HPN is a reasonable treatment option in patients with intestinal failure as a result of radiation enteritis. Survival and complications associated with HPN in patients with radiation enteritis seem to be similar to those in other HPN-treated groups. PMID- 11922562 TI - Lymphonodular hyperplasia of the terminal ileum associated with colitis shows an increase gammadelta+ t-cell density in children. AB - OBJECTIVE: Recently we reported a close association between lymphonodular hyperplasia (LNH) of the bulb of the duodenum and increased densities of intraepithelial gammadelta+ T-cells in subjects with untreated food allergies. In this study we sought to determine whether children with LNH of the terminal ileum (TI) show a similar correlation. METHODS: The mucosal specimens taken by colonoscopy from the TIs of 22 children with LNH of the TI without colitis, 13 with right-sided colitis or pancolitis, nine with left-sided colitis, eight with Crohn's disease, and three endoscopically healthy subjects were studied for T cell subsets with monoclonal antibodies using a three-layer peroxidase staining method. RESULTS: LNH of the TI was found in 32 of the 55 subjects (58%). In 22 it was the only endoscopic finding, but in nine of 13 subjects (69%) it was related to right-sided colitis or pancolitis. In patients with left-sided colitis or Crohn's disease it was diagnosed only rarely. In the whole study population, LNH of the of the TI showed a significant association with the increment in the density of gammadelta+ T-cells. The subjects with LNH of the TI and colitis starting from the cecum showed the highest values, discriminating them statistically from any other study group. Accordingly their gammadelta+/CD3+ ratio was high. Even in the subjects with LNH of the TI without colitis, the increment in gammadelta+ T-cells was significant as compared with the subjects with left-sided colitis. Upregulations of D-related expression on the mucosa of the TI were similar regardless of the presence of LNH or colitis or an increment in gammadelta+ T-cells. CONCLUSION: Our preliminary observations showed increased densities of intraepithelial gammadelta+ T-cells and elevated gammadelta+/CD3+ ratios in subjects with LNH on the mucosa of the TI, especially if related to colitis starting at the cecem, but not in subjects with typical left-sided colitis or granulomatous Crohn's disease. The study also provides further evidence suggesting the significance of food-borne antigens in the pathogenetic mechanism of right-sided colitis or pancolitis. The finding also indicates the significance of classifying colitis into gammadelta-positive and -negative diseases, and has implications for the treatment of these entities. PMID- 11922563 TI - Low free and total insulinlike growth factor I (IGF-I) and IGF binding protein-3 levels in chronic inflammatory bowel disease: partial normalization during prednisolone treatment. AB - OBJECTIVES: Catabolism and growth impairment are well known complications of chronic inflammatory bowel disease (CIBD). This may be caused by disease activity itself and/or the medical treatment, which may lead to changes in the growth hormone and insulinlike growth factor I (IGF-I) axis. Interest has focused on corticosteroids, as they are known to influence the growth hormone/IGF-I axis. METHODS: The aim of the present study was to examine changes in total and free IGF-I, IGF binding proteins (IGFBPs), and IGFBP-3 protease activity in 10 patients with severe acute activity in colon CIBD before and during high dose prednisolone treatment (1 wk) and 3 months tapering. Eight healthy subjects served as controls. RESULTS: Total and free IGF-I were significantly reduced by 35% and 53%, respectively, before prednisolone treatment (p < 0.05), and free IGF I remained reduced even by the end of the study period relative to controls (p < 0.05). IGFBP-3 was reduced by 16% before (p < 0.05), with normalization during prednisolone treatment and tapering relative to controls. There was no evidence of increase in IGFBP-3 protease activity. IGFBP-1 was increased before and tended also to be increased during prednisolone treatment and tapering. CONCLUSIONS: Marked changes in serum total and free IGF-I and IGFBPs were demonstrated in patients with severe exacerbation of CIBD without complete normalization during high dose prednisolone treatment and tapering. These changes may partly be involved in the catabolism of active CIBD patients. PMID- 11922564 TI - Synergistic inhibitory effect of cyclosporin A and vitamin D derivatives on T lymphocyte proliferation in active ulcerative colitis. AB - OBJECTIVE: 1,25-Dihydroxyvitamin D3 [1,25(OH)2D3], the hormonal active form of vitamin D3, could represent a potentially therapeutic agent in autoimmune diseases. Cyclosporin A (CsA) shows immunoregulatory properties, which, in many respects, seem to be similar to those of 1,25(OH)2D3. Our aim was to investigate the possible synergistic effect exerted by CsA in combination with 1,25(OH)2D3 or its nonhypercalcemic analogues, EB 1089 and KH 1060, on the proliferative response of T lymphocytes obtained from active ulcerative colitis patients. METHODS: The T lymphocyte-enriched population was treated with phytohemagglutinin and CsA (doses from 1 ng to 1000 ng/ml) alone or in association with 1,25(OH)2D3 or EB 1089 or KH 1060 (0.1, 1, 10 nM final concentration). Cell proliferation was determined by [3H]thymidine incorporation and analyzed on day 5 of culture. RESULTS: After incubation with CsA, T lymphocyte proliferation was significantly inhibited in comparison with the vehicle-treated cultures. However, T lymphocytes from ulcerative colitis patients were significantly more sensitive to CsA than those from healthy controls. The inhibition in T lymphocyte proliferation, after treatment of the cultures with CsA associated with either 1,25(OH)2D3 or EB 1089 or KH 1060, was synergistic at well-defined concentrations. CONCLUSIONS: Taking into account the lowest CsA dose (1 ng/ml), the highest synergistic inhibition in the proliferation of T lymphocytes prepared from ulcerative colitis patients was found combining CsA and 10 nM of 1,25(OH)2D3 or 10 nM of EB 1089 or KH 1060 at the three concentrations. The results obtained, associating the lowest CsA dose and the lowest KH 1060 concentration, may suggest an alternative therapeutic approach in these patients, reducing the dose, and consequently the toxicity, of CsA. PMID- 11922565 TI - HLA-DQ typing in the diagnosis of celiac disease. AB - OBJECTIVE: More than 95% of celiac patients share the major histocompatibility complex II class human leukocyte antigen (HLA) DQ2 or DQ8 haplotype; patients negative for both types are unlikely to suffer from celiac disease. Our aim was to investigate whether HLA-DQ2 and -DQ8 typing is helpful when diagnosis is uncertain because of the absence of unequivocal small bowel villous atrophy. METHODS: HLA-DQ2 and -DQ8 typing was carried out in 59 patients evincing nondiagnostic small bowel mucosal lesion or positive celiac serology, and in 17 patients maintaining a gluten-free diet without biopsy-proven celiac disease. HLA findings were compared to small bowel mucosal morphology; intraepithelial lymphocytes; and serum endomysial (EmA), reticulin, tissue transglutaminase (anti tTG) and gliadin antibodies. RESULTS: Of the 59 patients evincing only minor small bowel mucosal changes or positive celiac disease serology, 22 (37%) were negative for DQ2 and DQ8. All EmA-positive patients had celiac-type HLA, but antireticulin antibody, anti-tTg, and antigliadin antibody were also present in HLA-DQ2- and -DQ8-negative individuals. Eleven of 17 patients (65%) observing a gluten-free diet before small bowel biopsy did not share celiac-type HLA. None of the 17 had apparent villous atrophy. Serum EmA and anti-tTG were negative in all. HLA-DQ typing is less expensive than follow-up biopsy in the exclusion of celiac disease. CONCLUSIONS: HLA-DQ2 and -DQ8 determination is useful in exclusion, probably lifelong, of celiac disease in individuals with an equivocal small bowel histological finding. The low specificity of this test must, however, be borne in mind. PMID- 11922566 TI - Hepatitis C infection in African Americans: its natural history and histological progression. AB - OBJECTIVE: The aim of this retrospective analysis was to determine the natural history of hepatitis C virus infection in African Americans versus non-African Americans by evaluating the clinical, virological, and histological findings. METHODS: We examined in a retrospective manner the demographics, mode of infection, virological features, and histological progression of HCV infection in African Americans versus non-African Americans. There were 355 patients who met criteria based on adequate liver biopsy specimens and exclusion of other hepatic diseases. RESULTS: African Americans (n = 112) were significantly more likely to be infected with genotype 1 virus (88%) than were non-African Americans (n = 243; 67%; p < or = 0.001). Baseline HCV RNA levels were similar, although baseline ALT values were significantly lower in African Americans (80.0 microl +/- 5.5 vs 112.1 microl +/- 6.2; p < or = 0.001). African Americans were significantly older at the time of presentation and were significantly more likely to be women (p < or = 0.02). In African Americans, there was a trend toward less cirrhosis (22% vs 30%; p < or = 0.1) and significantly less piecemeal necrosis on liver biopsy. Non African Americans had significantly higher fibrosis scores, ALT values, and piecemeal necrosis ratings, and tended to progress more rapidly to cirrhosis. This difference in histological progression between the two groups was not explained by differences in alcohol consumption. CONCLUSION: The lower ALT, piecemeal necrosis scores, and slower progression of fibrosis in African Americans may reflect less immunological recognition of HCV-infected liver cells. PMID- 11922567 TI - Prevalence of autonomic dysfunction in cirrhotic and noncirrhotic portal hypertension. AB - OBJECTIVE: Autonomic dysfunction is common in patients with cirrhosis of the liver, but more so in patients with decompensated state, and is associated with increased mortality. We evaluated the presence and extent of autonomic dysfunction in patients with extrahepatic portal venous obstruction (EHPVO) and noncirrhotic portal fibrosis (NCPF), diseases with relatively preserved liver functions. METHODS: Heart rate variability in response to standing, deep breathing, and Valsalva maneuver and blood pressure response to sustained handgrip and standing were studied in 18 patients with EHPVO (13 mol/L, 5 F, mean age 15.2 +/- 6 yr), 12 patients with NCPF (5 mol/L, 7 F, mean age 26.4 +/- 8 yr), 15 patients with cirrhosis (7 mol/L, 8 F, mean age 12.6 +/- 6 yr), and 17 healthy controls (11 mol/L, 6 F, mean age 18.6 +/- 3 yr). Time-domain parameters of heart rate variability on 24-h ambulatory monitoring were assessed in all the patients. RESULTS: Autonomic dysfunction was observed in 67% of EHPVO, 25% of NCPF, and 80% of cirrhotic subjects but none of the healthy controls (p < 0.05). Four of five time-domain heart rate variability indices showed significant abnormalities in patients with EHPVO (p < 0.05) and cirrhosis (p < 0.05), when compared with patients with NCPF and healthy controls. CONCLUSIONS: Autonomic dysfunction is frequently encountered in patients with EHPVO and cirrhosis, and the presence of autonomic dysfunction in patients with noncirrhotic portal hypertension suggests a primary role of portal hypertension per se in the dysfunction. PMID- 11922568 TI - Tumor necrosis factor genetic polymorphisms and response to antiviral therapy in patients with chronic hepatitis C. AB - OBJECTIVE: Hepatitis C virus (HCV) is the major causal agent of non-A, non-B hepatitis and the leading indication for liver transplantation worldwide. The emerging field of immunogenetics has confirmed the significant role of heritability in host immune responses to infectious pathogens. Both the major and non-major histocompatibility complex genes are increasingly identified as candidate genes hypothesized to influence the susceptibility to, or the course of, a particular disease. We hypothesized that polymorphisms within the major histocompatibility complex class III region that encode for tumor necrosis factors (TNF)-alpha and TNF-beta might be predictive of response to antiviral therapy in patients with chronic hepatitis C. METHODS: A total of 155 subjects, including 110 HCV-seropositive individuals undergoing antiviral therapy and 45 ethnically similar HCV-negative controls, were studied. The HCV-positive patients had undergone antiviral treatment with either interferon monotherapy (n = 73) or in combination with ribavirin (n = 37) and were categorized as either nonresponders, sustained responders, or relapsers. Sixty (55%) patients had genotype 1 (1a or 1b). Genomic DNA was extracted, followed by polymerase chain reaction amplification and sequencing for two promoter TNF-alpha variants (at positions -238 and -308), as well as restriction fragment length analysis for four polymorphic loci within the TNF-beta gene (NcoI, TNFc, aa13, aa26). RESULTS: Although there was a trend toward higher frequency of the A allele in the TNF 238 promoter among HCV-infected patients (12% vs 4%), there were no significant differences in the distribution of the genotypic polymorphisms between patients and controls. Patients with the TNF 238 A allele had higher pretreatment viral loads as compared with patients homozygous for the wild type allele (7.2 x 10(6) +/- 4.2 x 10(6) copies/ml vs 3.8 x 10(6) +/- 0.34 x 10(6) copies/ml, p = 0.03). However, there was no association between TNF genetic markers, including multiple haplotypic combinations, and response to therapy. In addition, there was no correlation with these polymorphic loci and histological severity of liver disease. CONCLUSIONS: Although previous work has suggested potential roles for TNF in the pathogenesis of HCV infection, we were unable to identify any link between TNF genetic polymorphisms and histological severity or response to antiviral therapy. PMID- 11922569 TI - The cost-effectiveness of hepatitis A vaccination in patients with chronic hepatitis C viral infection in the United States. AB - OBJECTIVE: Hepatitis A vaccination is recommended for patients with chronic hepatitis C. Our aim was to analyze the cost-effectiveness of hepatitis A vaccination in these patients. The specific strategies evaluated were: no vaccination, targeted vaccination, and universal vaccination. METHODS: Clinical estimates were based on published data. Costs estimates were based on published data and institutional Medicare reimbursement rates. Health-related quality-of life weights were derived from published data and expert estimates. The target population consisted of patients 45 yr of age with chronic hepatitis C followed every 6 months until death. We adopted a societal perspective. RESULTS: Compared with no vaccination, targeted vaccination was associated with an incremental cost effectiveness ratio of $51,000 per quality-adjusted life-year. The incremental cost-effectiveness ratio of universal vaccination compared with targeted vaccination was $3,900,000 per quality-adjusted life-year. The results were particularly sensitive to the incidence of hepatitis A, probability of fulminant hepatic failure, and costs of hepatitis A antibody screening and vaccination. CONCLUSIONS: Targeted vaccination for hepatitis A in patients with chronic hepatitis C may be a cost-effective strategy to decrease the morbidity and mortality associated with hepatitis A superinfection. Universal vaccination is not a cost-effective alternative to targeted vaccination in this target population. PMID- 11922571 TI - Semiannual and annual surveillance of cirrhotic patients for hepatocellular carcinoma: effects on cancer stage and patient survival (Italian experience). AB - OBJECTIVES: Surveillance of cirrhotic patients for early detection of hepatocellular carcinoma, based on ultrasonography and alpha1-fetoprotein determination, is a recommended practice. However, it has not been proved that this procedure can improve patient survival. METHODS: We conducted a multicenter retrospective study on 1051 consecutive patients with hepatocellular carcinoma. The criteria for eligibility were presence of underlying cirrhosis, and description of cancer stage and modalities of its diagnosis. Among 821 patients fulfilling these criteria, the tumor was detected during semiannual surveillance in 215 individuals (group 1), during annual surveillance in 155 (group 2), and as a result of symptoms or incidentally in 451 (group 3). Survival of patients under surveillance was corrected for lead time. RESULTS: Cancer stage was similar in groups 1 and 2 and was less advanced than in group 3 (p < 0.001). The frequency of ablative treatments or chemoembolization was similar in groups 1 and 2 and was greater than in group 3 (p < 0.001). Both surveillance programs doubled the prevalence of potential candidates for liver transplantation (68.5% and 62.5%) with respect to group 3 (32.3%, p < 0.001). However, only 15 patients underwent transplantation. In groups 1 and 2, the 5-yr survival was equivalent and was greater than in group 3 (p < 0.001). By segregating patients according to severity of cirrhosis, the benefit was confined to compensated cirrhosis (adjusted relative risk of death for patients under surveillance: 0.59 [95% CI = 0.45-0.78]). CONCLUSIONS: Semiannual and annual surveillance equally improve the survival of cirrhotic patients with hepatocellular carcinoma and greatly increase the amenability rate to liver transplantation. When access to liver transplantation is limited, this benefit is restricted to patients with a good cirrhosis-related prognosis. PMID- 11922570 TI - Awareness of genetic testing for colorectal cancer predisposition among specialists in gastroenterology. AB - OBJECTIVES: Adult gastroenterologists practicing in New York State were surveyed to determine their practice with regard to identifying family histories consistent with inherited forms of colorectal cancer, and to assess their awareness of cancer genetic counseling and molecular genetic testing for familial adenomatous polyposis (FAP) and hereditary nonpolyposis colorectal cancer (HNPCC). METHODS: A closed-ended questionnaire was mailed to 815 gastroenterologists identified through the membership Directory of the American Gastroenterological Association (1998). Two mailings resulted in a response rate of 35%. RESULTS: In all, 99% of the gastroenterologists obtained a family history from their patients, and 95% were aware of cancer genetic counseling. However, only 51% would routinely refer patients for genetic counseling before providing cancer predisposition testing. In addition, only 52% were aware of the availability of genetic tests for FAP and 34% for HNPCC. Presented with a family history consistent with HNPCC, 79% could identify the syndrome, 26% recommended genetic counseling for the consultand, and 16% advised appropriate screening, according to current recommendations. CONCLUSIONS: The majority of gastroenterologists do obtain a family history on their patients. However, there is a need for physician education regarding the recognition of pedigrees consistent with inherited colorectal cancer, the genetic counseling process, and the availability of mutation testing for FAP and HNPCC. PMID- 11922572 TI - Selenoprotein levels in patients with colorectal adenomas and cancer. AB - OBJECTIVES: Selenium is a trace mineral that, as a constituent of certain selenoproteins, acts as an antioxidant. Results of studies addressing a cancer protective effect of selenium have been controversial. The present study measured selenoprotein-P, extracellular glutathione peroxidase, and plasma selenium in patients with colon cancer and adenomatous colon polyps to determine whether patients who develop colorectal adenomas or cancer are selenium deficient. METHODS: Patients who presented to an endoscopy center for colonoscopy or who were referred to our institution with a newly diagnosed colorectal cancer were offered enrollment in the trial. Each patient underwent phlebotomy, usually immediately after colonoscopy. In all, 103 patients were enrolled in the study. Of these, 33 patients were found to have colorectal cancer, 35 adenomatous colon polyps, and 17 normal examinations. A total of 18 patients had other diagnoses and were not included in the study group. RESULTS: The mean age for the colorectal cancer group was 69 yr, for the adenomatous colon polyp group 62 yr, and for the normal group was 56 yr. The adenomatous colon polyp and normal groups were predominantly female. Based on one way analysis of variance tests, there was no significant difference in selenoprotein-P or plasma selenium levels or extracellular glutathione peroxidase activity among the three groups (p = 0.28, 0.098, and 0.35 respectively). CONCLUSIONS: The present data suggest that patients with adenomatous colon polyps and those with colorectal cancer are not selenium deficient. PMID- 11922573 TI - Cased surgical set. PMID- 11922574 TI - Extraction, isolation, and SDS-PAGE analysis of purified gastric mucin in a patient with Menetrier's disease. AB - Menetrier's disease is a rare condition characterized by marked proliferation of gastric mucosa with variable mucus secretion and achlorhydria. Although crude mucus secretion and gastric aspirates have been evaluated in this disease for output of dry matter, hexosamine, fucose, protein content, and transforming growth factor alpha activity, we report for the first time the isolation, purification, and gel electrophoresis of mucin from crude mucus scrapings. The fragmentation pattern of mucin in Menetrier's disease demonstrated less large polymeric mucin than the control. There was also a band of approximately 55-65 kd M, on polyacrylamide gel electrophoresis similar to that found in gastric carcinoma or peptic ulcer, but absent in the control specimens. PMID- 11922575 TI - PPIs meet the liver. PMID- 11922576 TI - "Hot spots" can burn you. PMID- 11922577 TI - Reevaluation of colon cancer staging--stricture presence reflects advanced disease? PMID- 11922578 TI - Endoscopic markers of celiac disease. PMID- 11922579 TI - Re: Fefferman et al.--"Recurrence" of chronic pancreatitis appearing on a patient with underlying Crohn's disease. PMID- 11922580 TI - A case of Peutz-Jeghers syndrome associated with duodenal carcinoma and sickle cell anemia. PMID- 11922581 TI - Cerebral blood flow in basal ganglia is correlated with clinical signs of hepatic encephalopathy in patients with liver cirrhosis. PMID- 11922582 TI - A pilot study to evaluate stomach cancer risk reduction by Helicobacter pylori eradication. PMID- 11922583 TI - Needle track metastasis of primary liver carcinoma after percutaneous needle aspiration. PMID- 11922584 TI - Lactoferrin inhibits hepatitis C virus viremia in chronic hepatitis C patients with high viral loads and HCV genotype 1b. PMID- 11922585 TI - Autoimmune hepatitis and interferon beta-1a for multiple sclerosis. PMID- 11922586 TI - Acquired and hereditary thrombotic risk factors in patients with acute mesenteric vein thrombosis. PMID- 11922587 TI - Duodenoscopic diagnosis of perforated periampullary diverticulitis. PMID- 11922588 TI - Gastric ulcer as a rare complication of percutaneous ethanol injection for hepatocellular carcinoma. PMID- 11922589 TI - Massive steatosis hepatis: an unusual manifestation of Whipple's disease. PMID- 11922590 TI - Usefulness of endoscopic appearance for choosing a biopsy target site and determining complete remission of primary gastric lymphoma of mucosa-associated lymphoid tissue after eradication of Helicobacter pylori infection. PMID- 11922592 TI - Severe cholestatic hepatitis in a patient taking acitretin. PMID- 11922591 TI - Spontaneous regression of hepatocellular carcinoma associated with elevated levels of interleukin 18. PMID- 11922593 TI - Sequential solubilization of proteins precipitated with trichloroacetic acid in acetone from cultured Catharanthus roseus cells yields 52% more spots after two dimensional electrophoresis. AB - Sample preparation is still the most critical step in two-dimensional gel electrophoresis (2-DE), and needs to be optimized for each type of sample. To analyze the proteome of the medicinal plant Catharanthus roseus, we developed and evaluated a sequential solubilization procedure for the solubilization of proteins after precipitation in trichloroacetic acid and acetone. The procedure includes solubilization with a conventional urea buffer followed by a stronger solubilizing buffer containing thiourea. The sequential solubilization of the precipitated proteins results in very different spot patterns following 2-DE. The number of protein spots which could be detected in both samples of the sequential solubilization was only about 10% of the total number of spots. Compared to solubilization in a single step, the total number of spots that could be detected in the sequential solubilization procedure was increased by 52%. The method described is simple and is applicable to different types of plant tissue. PMID- 11922594 TI - Histometrics: improvement of the dynamic range of fluorescently stained proteins resolved in electrophoretic gels using hyperspectral imaging. AB - Most image-based analyses, using absorbance or fluorescence of the spatial distribution of identifiable structures in complex biological systems, use only a very small number of dimensions of possible spectral data for the generation and interpretation of the image. We here extend the concepts of hyperspectral imaging, being developed in remote sensing, into analytical biotechnology. The massive volume of information contained in hyperspectral spectroscopic images requires multivariate analysis in order to extract the chemical and spatial information contained within the data. We here describe the use of multivariate statistical methods to map and quantify common protein staining fluorophores (SYPRO Red, Orange and Tangerine) in electrophoretic gels. Specifically, we find (a) that the 'background' underpinning limits of detection is due more to proteins that have not migrated properly than to impurities or to ineffective destaining, (b) the detailed mechanisms of staining of SYPRO red and orange are apparently not identical, and in particular (c) that these methods can provide two orders of magnitude improvement in the detection limit per pixel, to levels well below the limit observable optically. PMID- 11922595 TI - Improved silver staining protocols for high sensitivity protein identification using matrix-assisted laser desorption/ionization-time of flight analysis. AB - In proteomics it is essential to be able to detect proteins separated by gel electrophoresis at high sensitivity. Silver staining is currently the most popular method. Here we present silver staining protocols that are optimized for staining sensitivity, peptide recovery and compatibility with digestion and mass spectrometry. PMID- 11922596 TI - Improvement of an in-gel tryptic digestion method for matrix-assisted laser desorption/ionization-time of flight mass spectrometry peptide mapping by use of volatile solubilizing agents. AB - The combination of matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS), in-gel enzymatic digestion of proteins separated by two-dimensional gel electrophoresis and searches of molecular weight in peptide-mass databases is a powerful and well established method for protein identification in proteomics analysis. For successful protein identification by MALDI-TOF mass spectrometry of peptide mixtures, critical parameters include highly specific enzymatic cleavage, high mass accuracy and sufficient numbers and sequence coverage of the peptides which can be analyzed. For in-gel digestion with trypsin, the method employed should be compatible both with enzymatic cleavage and subsequent MALDI-TOF MS analysis. We report here an improved method for preparation of peptides for MALDI-TOF MS mass fingerprinting by using volatile solubilizing agents during the in-gel digestion procedure. Our study clearly demonstrates that modification of the in-gel digestion protocols by addition of dimethyl formamide (DMF) or a mixture of DMF/N,N-dimethyl acetamide at various concentrations can significantly increase the recovery of peptides. These higher yields of peptides resulted in more effective protein identification. PMID- 11922597 TI - Bioinformatic assessment of mass spectrometric chemical derivatisation techniques for proteome database searching. AB - Identification of proteins from the mass spectra of peptide fragments generated by proteolytic cleavage using database searching has become one of the most powerful techniques in proteome science, capable of rapid and efficient protein identification. Using computer simulation, we have studied how the application of chemical derivatisation techniques may improve the efficiency of protein identification from mass spectrometric data. These approaches enhance ion yield and lead to the promotion of specific ions and fragments, yielding additional database search information. The impact of three alternative techniques has been assessed by searching representative proteome databases for both single proteins and simple protein mixtures. For example, by reliably promoting fragmentation of singly-charged peptide ions at aspartic acid residues after homoarginine derivatisation, 82% of yeast proteins can be unambiguously identified from a single typical peptide-mass datum, with a measured mass accuracy of 50 ppm, by using the associated secondary ion data. The extra search information also provides a means to confidently identify proteins in protein mixtures where only limited data are available. Furthermore, the inclusion of limited sequence information for the peptides can compensate and exceed the search efficiency available via high accuracy searches of around 5 ppm, suggesting that this is a potentially useful approach for simple protein mixtures routinely obtained from two-dimensional gels. PMID- 11922598 TI - Combined automated PCR cloning, in vitro transcription/translation and two dimensional electrophoresis for bacterial proteome analysis. AB - The most popular approach for proteomics analysis is based on the combination of two-dimensional gel electrophoresis and mass spectrometry (MS). Although very effective, the method suffers from a number of limitations, the most serious one being the necessity to have expensive and sophisticated instrumentation requiring handling by skilled personnel. Here we propose an alternative approach which may offer some advantages over the current methods, at least for some specific applications. The method is based on two-dimensional gel separation of radiolabeled synthetic proteins derived from transcription/translation reactions of linear polymerase chain reaction amplified genes. The gel is autoradiographed and this is superimposed on the sample gel whose protein spots have to be identified. Matching between autoradiographs and sample gel spots allows immediate protein identification. The method has been validated identifying six proteins from a membrane protein preparation of Neisseria meningitidis MC58 strain. All proteins were correctly identified as judged by confirmation analysis with MS. The approach is particularly useful when a specific subset of proteins needs to be identified in a complex protein mixture. PMID- 11922599 TI - Isolation and proteomic characterization of the major proteins of the nucleolin binding ribonucleoprotein complexes. AB - Nucleolin (NCL) is one of the most abundant nucleolar proteins of exponentially growing eukaryotic cells. It is known to interact only transiently with rRNA and preribosomal particles and not to be detectable in mature cytoplasmic ribosomes, and is believed to function as multi-protein complexes during ribosome biogenesis and maturation. However, those multiprotein complexes remain only partially characterized due to the difficulty of conventional protein analysis methods. Here we report isolation of NCL-binding protein complex and its proteomic characterization with the use of an analytical method based on matrix-assisted laser desorption/ionization-time of flight analysis coupled with searching peptide mass databases. The NCL-binding protein complex was isolated by immunoprecipitation with anti-Flag antibody from human kidney 293 cells that were transfected with the Flag-tagged NCL gene, and showed RNA integrity for holding their protein constituents. Interaction between NCL and its binding complex was disrupted by an RNA oligonucleotide with a NCL recognition element, indicating that NCL binds to the ribonucleoprotein (RNP) complex mainly through the sequence specific protein-RNA interaction. We confirmed that an RNA-binding domain of NCL alone was sufficient to hold the entire NCL-binding RNP complex, indicating the strict binding specificity of NCL to the isolated RNP complex in 293 cells. We identified forty ribosomal proteins from both the large and small subunits, and twenty nonribosomal proteins. These results together suggest that the isolated NCL-binding RNP complex is a preribosomal particle present in the nucleolus of 293 cells. PMID- 11922600 TI - Identification and mapping of human saphenous vein medial smooth muscle proteins by two-dimensional polyacrylamide gel electrophoresis. AB - Changing smooth muscle phenotype and abnormal cell proliferation are important features of vascular pathology, including the failure of saphenous vein bypass grafts. We have characterised and mapped protein expression in human saphenous vein medial smooth muscle, using two-dimensional (2-D) polyacrylamide gel electrophoresis. The 2-D system comprised a nonlinear immobilised pH 3-10 gradient in the first dimension (separating proteins with isoelectric point values between pH 3-10), and 12%T total gel concentration sodium dodecyl sulphate polyacrylamide gel electrophoresis in the second dimension (separating proteins in the range 14,000-200,000 Daltons). Using a combination of peptide mass fingerprinting by matrix-assisted laser desorption/ionisation-time of flight mass spectrometry and partial amino acid sequencing by nanospray tandem mass spectrometry, a subset of 149 protein spots was analysed, with 129 protein spots being identified and mapped. The data presented here are an important addition to the limited knowledge of venous medial smooth muscle protein expression in vivo. Our protein map will facilitate the identification of proteins differentially expressed in human saphenous vein bypass grafts. In turn, this may lead to the elucidation of molecular events involved in saphenous vein bypass graft failure. The map should also provide a basis for comparative studies of protein expression in vascular smooth muscle of varying origins. PMID- 11922601 TI - Oncoprotein 18 levels and phosphorylation mediate megakaryocyte polyploidization in human erythroleukemia cells. AB - Megakaryocytes undergo an unusual cell cycle during differentiation that results in polyploidy through largely unknown mechanism(s). It has been shown that serine phosphorylation of oncoprotein 18 (Op18) is required for cell cycle progression specifically at the G2/M transition. Moreover, mutant forms of Op18 that are defective in one or more of the four serine residues induce G2/M arrest and subsequent polyploidization. Op18 phosphorylation is rapidly induced with phorbol myristate acetate (PMA) treatment in a wide range of human cells. In this study, we investigated the role of Op18 in PMA induced polyploidization during megakaryocyte differentiation of the human erythroleukemia cell line. Crucial to the molecular analysis of megakaryocyte differentiation, is the ability to fractionate cell populations with different ploidy levels. We have utilized cell elutriation as a fractionation strategy to analyze Op18 expression in synchronized cell subpopulations in different phases of the cell cycle or with progressive megakaryocyte polyploidization. In the absence of PMA, increased phosphorylation of Op18 was observed in HEL cells during cell cycle progression, as for other proliferating cells. However, in contrast to Jurkat leukemia cells chosen as control, HEL cells exhibited a lack of Op18 phosphorylation in response to PMA, which was accompanied by polyploidization and differentiation along the megakaryocytic lineage. To further determine the role of Op18 in polyploidization, HEL cells were transfected with different Op18 expression constructs. Differences in cell survival and polyploidization were observed between high and low Op18 expressors. An increased Op18 level reduced cell survival during the early stage of PMA induced megakaryocyte differentiation, but enhanced polyploidization efficiency. Our findings suggest that maintenance of a high level of unphosphorylated Op18 is required for efficient polyploidization during the differentiation program of megakaryocytes. PMID- 11922602 TI - Establishment of a root proteome reference map for the model legume Medicago truncatula using the expressed sequence tag database for peptide mass fingerprinting. AB - We have established a proteome reference map for Medicago truncatula root proteins using two-dimensional gel electrophoresis combined with peptide mass fingerprinting to aid the dissection of nodulation and root developmental pathways by proteome analysis. M. truncatula has been chosen as a model legume for the study of nodulation-related genes and proteins. Over 2,500 root proteins could be displayed reproducibly across an isoelectric focussing range of 4-7. We analysed 485 proteins by peptide mass fingerprinting, and 179 of those were identified by matching against the current M. truncatula expressed sequence tag (EST) database containing DNA sequences of approximately 105,000 ESTs. Matching the EST sequences to available plant DNA sequences by BLAST searches enabled us to predict protein function. The use of the EST database for peptide identification is discussed. The majority of identified proteins were metabolic enzymes and stress response proteins, and 44% of proteins occurred as isoforms, a result that could not have been predicted from sequencing data alone. We identified two nodulins in uninoculated root tissue, supporting evidence for a role of nodulins in normal plant development. This proteome map will be updated continuously (http://semele.anu.edu.au/2d/2d.html) and will be a powerful tool for investigating the molecular mechanisms of root symbioses in legumes. PMID- 11922603 TI - Delineation of in vivo assembled multiprotein complexes via biomolecular interaction analysis mass spectrometry. AB - Biomolecular interaction analysis mass spectrometry (BIA-MS) is a multiplexed bioanalytical approach used in analysis of proteins from complex biological mixtures. It utilizes surface-immobilized ligands for protein affinity retrieval, surface plasmon resonance for monitoring the ligand-protein interaction and matrix-assisted laser desorption/ionization-time of flight mass spectrometry for revealing the masses of the biomolecules retrieved by the ligand. In order to explore the utility of BIA-MS in delineation of multiprotein complexes, an in vivo assembled protein complex comprised of retinol binding protein (RBP) and transthyretin (TTR) was investigated. Antibodies to RBP and TTR were utilized as ligands in the analysis of the protein complex present in human plasma. The RBP TTR complex was retrieved by the anti-RBP antibody as indicated by the presence of both RBP and TTR signals in the mass spectra. RBP signals were not observed in the mass spectra of the material retained on the anti-TTR derivatized surface. In addition, the mass-specific detection in BIA-MS allowed detection of RBP and TTR analyte variants. PMID- 11922604 TI - African swine fever virus-induced polypeptides in porcine alveolar macrophages and in Vero cells: two-dimensional gel analysis. AB - High-resolution two-dimensional electrophoresis followed by computer analysis has been used to study quantitatively the patterns of protein synthesis produced in porcine alveolar macrophages and in Vero cells infected with African swine fever virus (ASFV). Initially, a protein database for each cell type was constructed. The porcine alveolar macrophage database includes 995 polypeptides (818 acidic, isoelectric focusing (IEF) and 177 basic, nonequilibrium pH gradient electrophoresis (NEPHGE)) whereas the Vero database contains 1,398 polypeptides (1,127 acidic, IEF and 271 basic, NEPHGE). Taking these databases as reference, ASFV highly virulent strain E70 induces 57 acid and 43 basic polypeptides in porcine alveolar macrophages, which account for most of the information content of the virus DNA. The kinetics of synthesis of the virus-induced polypeptides showed the existence of three classes of proteins: one whose synthesis starts early after infection, continues for a period and then switches off; another whose synthesis also starts early but continues for prolonged periods; and a third which requires DNA replication. The attenuated, cell adapted, strain BA71V induces 92 acidic and 37 basic proteins in Vero cells. Significant differences were observed when comparing the patterns of polypeptides induced by the two viral strains. In both cell systems studied, ASFV infection produces a general shutoff of protein synthesis that affects up to 65% of the cellular proteins. Interestingly, 28 proteins of porcine alveolar macrophages and 48 proteins of Vero cells are stimulated at least two times by ASFV infection. PMID- 11922605 TI - Comparative proteomics to establish genetic relationships in the Brassicaceae family. AB - The comparisons of the same proteome (aerial part of etiolated seedlings) in different species and genus of the plant family Brassicaceae (cabbages, mustards, rapes, radishes and Arabidopsis) was undertaken to establish genetic proximities between them. Genetic distances were calculated on the basis of common and distinct spots. The construction of phenetic trees and factorial correspondence analysis that were performed are in very good agreement with our actual knowledge of the taxonomy of the Brassicaceae. The position of the tetraploids, for instance, is in between the positions of their constitutive genome donor representatives. Comparative proteomics may be helpful to shorten the transfer between model and agronomic target species. PMID- 11922606 TI - Proteomic identification of glutathione S-transferases from the model nematode Caenorhabditis elegans. AB - Glutathione affinity chromatography and two-dimensional electrophoresis (2-DE) were used to purify glutathione binding proteins from Caenorhabditis elegans. All proteins identified after peptide mass fingerprinting using matrix-assisted laser desorption/ionization-time of flight were found to belong to the glutathione S transferase (GST) superfamily. From the 26 individual spots identified, 12 different GSTs were isolated. Of these, five were found on the gel only once, whilst the remaining seven were represented by 21 separate spots. Most of the GSTs identified were of the nematode specific class, however, three Alpha class GSTs, a Pi and a Sigma class GST were also isolated. PMID- 11922607 TI - The SELDI-TOF MS approach to proteomics: protein profiling and biomarker identification. AB - The need for methods to identify disease biomarkers is underscored by the survival-rate of patients diagnosed at early stages of cancer progression. Surface enhanced laser desorption/ionization time-of-flight mass spectrometry (SELDI-TOF MS) is a novel approach to biomarker discovery that combines two powerful techniques: chromatography and mass spectrometry. One of the key features of SELDI-TOF MS is its ability to provide a rapid protein expression profile from a variety of biological and clinical samples. It has been used for biomarker identification as well as the study of protein-protein, and protein-DNA interaction. The versatility of SELDI-TOF MS has allowed its use in projects ranging from the identification of potential diagnostic markers for prostate, bladder, breast, and ovarian cancers and Alzheimer's disease, to the study of biomolecular interactions and the characterization of posttranslational modifications. In this minireview we discuss the application of SELDI-TOF MS to protein biomarker discovery and profiling. PMID- 11922608 TI - Cloning and characterization of a proliferation-associated cytokine-inducible protein, CIP29. AB - We identified a novel erythropoietin (Epo)-induced protein (CIP29) in lysates of human UT-7/Epo leukemia cells using two-dimensional gel analysis and cloned its full-length cDNA. CIP29 contains 210 amino acids with a predicted MW of 24 kDa, and has a N-terminal SAP DNA-binding motif. CIP29 expression was higher in cancer and fetal tissues than in normal adult tissues. CIP29 mRNA expression is cytokine regulated in hematopoietic cells, being up-regulated by Epo in UT7/Epo cells, and by thrombopoietin (Tpo), FLT3 ligand (FL) and stem cell factor (SCF) in primary human CD34(+) cells. Up-regulation of CIP29 in UT7/Epo cells by Epo was associated with cell cycle progression but not with antiapoptosis. Epo withdrawal reduced CIP29 expression concomitant with cell cycle arrest. Overexpression of CIP29-GFP in HEK293 cells enhances cell cycle progression. CIP29 appears to be a new cytokine regulated protein involved in normal and cancer cell proliferation. PMID- 11922609 TI - Identification and characterization of Streptococcus pneumoniae Ffh, a homologue of SRP54 subunit of mammalian signal recognition particle. AB - Recent studies have demonstrated that bacteria possess an essential protein translocation system similar to mammalian signal recognition particle (SRP). Here we have identified the Ffh, a homologue of the mammalian SRP54 subunit from S. pneumoniae. Ffh is a 58-kDa protein with three distinct domains: an N-terminal hydrophilic domain (N-domain), a G-domain containing GTP/GDP binding motifs, and a C-terminal methionine-rich domain (M-domain). The full-length Ffh and a truncated protein containing N and G domains (Ffh-NG) were overexpressed in E. coli and purified to homogeneity. The full-length Ffh has an intrinsic GTPase activity with k(cat) of 0.144 min(-1), and the K(m) for GTP is 10.9 microM. It is able to bind to 4.5S RNA specifically as demonstrated by gel retardation assay. The truncated Ffh-NG has approximately the same intrinsic GTPase activity to the full-length Ffh, but is unable to bind to 4.5S RNA, indicating that the NG domain is sufficient for supporting intrinsic GTP hydrolysis, and that the M domain is required for RNA binding. The interaction of S. pneumoniae Ffh with its receptor, FtsY, resulted in a 20-fold stimulation in GTP hydrolysis. The stimulation was further demonstrated to be independent of the 4.5S RNA. In addition, a similar GTPase stimulation is also observed between Ffh-NG and FtsY, suggesting that the NG domain is sufficient and the M domain is not required for GTPase stimulation between Ffh and FtsY. PMID- 11922611 TI - Cloning and tissue distribution of three murine alpha/beta hydrolase fold protein cDNAs. AB - We have cloned 3 novel murine cDNAs encoding proteins containing an alpha/beta hydrolase fold; a catalytic domain found in a very wide range of enzymes. These proteins belong to the prosite UPF0017 uncharacterized protein family and we have named them lung alpha/beta hydrolase 1, 2, and 3 (LABH) since they were cloned from lung cDNA. All have 9 coding exons, encoding 412, 425, and 411 residue proteins respectively (46-48 kDa); LABH1 being closely related to LABH3 having 45% identity. All 3 proteins have a single predicted amino-terminus transmembrane domain. An alignment of family members from different phyla enabled the identification of the LABH1 catalytic triad as Ser211, Asp337, and His366. mRNA expression levels of LABH1 and 3 were highest in liver and LABH2 highest in testis. These findings suggest that the LABH proteins consist of a novel family of membrane bound enzymes whose function has yet to be determined. PMID- 11922610 TI - EVI1 is expressed in megakaryocyte cell lineage and enforced expression of EVI1 in UT-7/GM cells induces megakaryocyte differentiation. AB - 3q21q26 syndrome, an acute myeloid leukemia (AML) or myelodysplastic syndrome (MDS) with chromosomal translocations or inversions between the bands 3q21 and 3q26, is frequently associated with dysmegakaryocytopoiesis and increased platelet counts at the initial diagnosis. Since the EVI1 gene at 3q26 is transcriptionally activated in 3q21q26 syndrome, we assessed the role of EVI1 gene expression in the abnormal megakaryocytic differentiation in 3q21q26 syndrome. RT-PCR analysis of various types of hematopoietic cells revealed that the EVI1 gene is expressed specifically in CD34(+) cells, megakaryocytes, and platelets. UT-7 is a human immature megakaryoblastic leukemia cell line with dependence for the growth on granulocyte-macrophage colony-stimulating factor (GM CSF) (designated at UT-7/GM) and with a differentiation capacity to erythroid (UT 7/EPO) and megakaryocytic lineages (UT-7/TPO) by erythropoietin (EPO) and thrombopoietin (TPO), respectively. Among three UT-7 sublines, UT-7/GM, UT-7/EPO, and UT-7/TPO, expression of the EVI1 gene was detected at low levels in UT-7/GM and UT-7/EPO cells, but was detected at a higher level in UT-7/TPO cells. When UT 7/GM cells were cultured with TPO, the level of EVI1 expression was increased, along with increased numbers of polynuclear megakaryocytes and expression of the platelet factor 4 (PF-4) gene. Furthermore, forced expression of the EVI1 gene in UT-7/GM cells changed their morphology to polynuclear megakaryocytes, stopped their growth, and induced cell death within a month. These data indicate that expression of the EVI1 gene is involved in progression of megakaryocytic differentiation and, thus, the dysmegakaryocytopoiesis in 3q21q26 syndrome could be partly due to an enhanced differentiation capacity of leukemia cells and/or megakaryocytes by constitutive expression of the EVI1 gene. PMID- 11922612 TI - The short MCK1350 promoter/enhancer allows for sufficient dystrophin expression in skeletal muscles of mdx mice. AB - First-generation adenovirus vectors (AdV) have been used successfully to transfer a human dystrophin minigene to skeletal muscle of mdx mice. In most studies, strong viral promoters such as the cytomegalovirus promoter/enhancer (CMV) were used to drive dystrophin expression. More recently, a short version of the muscle creatine kinase promoter (MCK1350) has been shown to provide muscle-specific reporter gene expression after AdV-mediated gene delivery. Therefore, we generated a recombinant AdV where dystrophin expression is controlled by MCK1350 (AdVMCKdys). AdVMCKdys was injected by the intramuscular route into anterior tibialis muscle of mdx mice shortly after birth. Dystrophin expression was assessed at 20, 30, and 60 days after AdV-injection. At 20 days, muscles of AdVMCKdys-injected mdx mice showed a high number of dystrophin-positive fibers (mean: 365). At 60 days, the number of dystrophin-positive fibers was not only maintained, but increased significantly (mean: 600). In conclusion, MCK1350 allows for sustained dystrophin expression after AdV-mediated gene transfer to skeletal muscle of newborn mdx mice. In contrast to previous studies, where strong viral promoters were used, dystrophin expression driven by MCK1350 peaks at later time points. This may have implications for the future use of muscle specific promoters for gene therapy of Duchenne muscular dystrophy. PMID- 11922613 TI - Intestinal metabolism and absorption of cholecystokinin analogs in rats. AB - Intestinal metabolism and poor permeability were known to be major barriers for oral absorption of large peptide drugs. Dimensionless wall permeability values of C-terminal octa- and tetra-peptides cholecystokinin analogs (CCK8 and CCK4) were estimated and found out to be greater than 1, suggesting no permeability-limited absorption for CCK analogs. Thus, a strategy employing enzyme inhibitors and a specific delivery site to improve the absorption was developed and tested with CCK8, followed by identification of metabolites of the analogs and their participating enzymes in rabbit brush-border membrane vesicles. Thiorphan and amastatin, a specific enzyme inhibitor for enkephalinase and aminopeptidase, respectively, in pH 4 buffer solution were coadministered with CCK8 to the ileum in fistulated rats. The absolute bioavailability (F) of CCK8 was 5.4% and increased to 19% in the presence of the enzyme inhibitors, while the F values following oral administration were close to zero. These results indicate that peptide oral delivery is possible. PMID- 11922614 TI - Role of tryptophan hydroxylase phe313 in determining substrate specificity. AB - The active site residue phenylalanine 313 is conserved in the sequences of all known tryptophan hydroxylases. The tryptophan hydroxylase F313W mutant protein no longer shows a preference for tryptophan over phenylalanine as a substrate, consistent with a role of this residue in substrate specificity. A tryptophan residue occupies the homologous position in tyrosine hydroxylase. The tyrosine hydroxylase W372F mutant enzyme does not show an increased preference for tryptophan over tyrosine or phenylalanine, so that this residue cannot be considered the dominant factor in substrate specificity in this family of enzymes. PMID- 11922615 TI - Shikonin stimulates glucose uptake in 3T3-L1 adipocytes via an insulin independent tyrosine kinase pathway. AB - Type 2 diabetes is due to defects in both insulin action and secretion. In an attempt to discover small molecules that stimulate glucose uptake, similar to insulin, a cell-based glucose uptake screening assay was performed using 3T3-L1 adipocytes. Shikonin, a substance originally isolated from the root of the Chinese plant that has been used as an ointment for wound healing, was thus identified. Shikonin stimulated glucose uptake and potentiated insulin-stimulated glucose uptake in a concentration-dependent manner in 3T3-L1 adipocytes. Stimulation of glucose uptake was also observed in rat primary adipocytes and cardiomyocytes. Like insulin, shikonin-stimulated glucose uptake was inhibited by genistein, a tyrosine kinase inhibitor, and enhanced by vanadate, a tyrosine phosphatase inhibitor. However, in contrast to insulin, shikonin-stimulated glucose uptake was not strongly inhibited by wortmannin, a specific inhibitor of phosphatidylinositol 3-kinase (PI3K). In vitro phosphorylation analyses revealed that shikonin did not induce tyrosine phosphorylation of the insulin receptor, but significantly induced both Thr-308 and Ser-473 phosphorylation of Akt. Our results suggest that in 3T3-L1 adipocytes, shikonin action is not mediated primarily via the insulin receptor/PI3K pathway, but rather via another distinct tyrosine kinase-dependent pathway leading to glucose uptake involving Akt phosphorylation. PMID- 11922616 TI - Metal-catalyzed oxidation of extracellular matrix proteins promotes human mesangial cell apoptosis and is associated with enhanced expression of Bax and caspase activation. AB - Oxidative injury in glomerular disease may oxidize extracellular matrix proteins which might modulate their interaction with mesangial cells and thereby account for the hypocellularity seen in advanced glomerulosclerosis. In this study we evaluated whether oxidation of extracellular matrix could modulate human mesangial cell apoptosis. Human mesangial cells were seeded onto plates coated with unmodified (control) or oxidized Matrigel, laminin, or type IV collagen. Mesangial cell apoptosis was increased on oxidized Matrigel as well as on oxidized laminin and type IV collagen. Mesangial cells behaved similarly on plates coated with control and oxidized forms of the integrin ligand-containing peptide GRGDSP. Cells on oxidized matrix demonstrated enhanced expression of Bax, increased fragmentation of PARP, and diminished apoptosis in the presence of the interleukin-1 beta converting enzyme inhibitor Ac-Tyr-Val-Ala-Asp-aldehyde. These data suggest that oxidation of extracellular matrix proteins may enhance human mesangial cell apoptosis via a mechanism that appears to involve enhanced expression of Bax and caspase activation. This may account for irreversible mesangial hypocellularity in glomerulosclerosis. PMID- 11922617 TI - Human eukaryotic initiation factor 4AII associates with hepatitis C virus NS5B protein in vitro. AB - Hepatitis C virus (HCV) NS5B protein has been shown to have RNA-dependent RNA polymerase (RdRp) activity by itself and is a key enzyme involved in viral replication. Using analyses with the yeast two-hybrid system and in vitro binding assay, we found that human eukaryotic initiation factor 4AII (heIF4AII), which is a component of the eIF4F complex and RNA-dependent ATPase/helicase, interacted with NS5B protein. These two proteins were shown to be partially colocalized in the perinuclear region. The binding site in HCV NS5B protein was localized within amino acid residues 495 to 537 near the C terminus. Since eIF4A has a helicase activity and functions in a bidirectional manner, the binding of HCV NS5B protein to heIF4AII raises the possibility that heIF4AII facilitates the genomic RNA synthesis of NS5B protein by unwinding the secondary structure of the HCV genome and is a host component of viral replication complex. PMID- 11922618 TI - Expression and function of NJ-1 surface antigen in megakaryopoiesis. AB - Immunostaining with NJ-1 monoclonal antibody (MoAb) revealed that NJ-1 is expressed on megakaryocytes (MKs). NJ-1-positive and lineage-negative progenitor cells have a higher potency to proliferate and differentiate into MKs. MKs were divided into NJ-1(+)MKs and NJ-1(-)MKs. NJ-1(+)MKs are immature MKs because of their low potential to generate pro-platelets. When cultured CD41-positive MK cells were analyzed with RT-PCR, we found that the expression of NJ-1 is down regulated. NJ-1(+)MKs have a high adherent potential to endothelial cells comparing with NJ-1(-)MKs, and this binding ability could be inhibited by the NJ 1-Fc fusion protein. We hypothesize that NJ-1(+)MKs are immature MKs and the NJ-1 molecule is involved in MK adhesion to endothelial cells. PMID- 11922619 TI - Apoptosis and related proteins during parturition in prostaglandin F receptor deficient mice. AB - This study investigated whether apoptosis and related proteins are involved in parturition by comparative observation of FP-deficient mice without labor and wild type mice with vaginal delivery. We examined the expression of apoptosis, Fas, FasL, active caspase-3 and bcl-2 proteins in the amnion, placenta and decidua. DNA laddering in the amnion, placenta and decidua tissue did not significantly differ between FP-deficient and wild type mice on day 18 of pregnancy. Similar TUNEL staining results were found in all tissues of FP deficient mice compared with those of wild type mice. A higher intensity of apoptotic cells was found in the decidua basalis. The index of TUNEL-positive cells were not significantly different in the amnion, placenta and decidua of FP deficient mice compared with that of wild type mice on day 18 of pregnancy. Specific bands for Fas were clearly observed in the amnion, placenta and decidua tissue. FasL specific bands were observed in the placenta and decidua, but a few in amnion tissue. A great number of active caspase-3 specific bands were detected in decidua, while a few such bands were detected in the placenta and few bands in the amniotic tissue. Bands for bcl-2 were detected in the amnion, placenta and decidua tissue. The weakest band was in decidual tissue. Fas, FasL, active caspase-3, and bcl-2 specific bands did not show any significant differences between the two groups. These findings demonstrate that apoptosis, Fas, FasL, caspase-3, and Bcl-2 occur in mouse term placenta that is not involved in parturition. PMID- 11922620 TI - Investigation of antigen delivery route in vivo and imune-boosting effects mediated by pH-sensitive liposomes encapsulated with K(b)-restricted CTL epitope. AB - Using fluorescein isothiocyanate (FITC)-conjugated H-2K(b) CTL epitope (SIINFEKL) as a model system, we investigated the antigen delivery route by pH-sensitive liposomes in vivo. Fluorescence was initially detected in lymph nodes at 3 h after immunization, and its intensity reached a peak value in superticial inguinal lymph node at 9 h. No trace could be detected in spleen even with prolonged monitoring for up to 24 h. These results strongly suggest that the presentation of CTL-peptide antigen vehicled by pH-sensitive liposomes exclusively occurs in lymph nodes. In mice immunized with the H-2K(b) CTL epitope encapsulated pH-sensitive liposomes, peptide-specific CTL response was detected at day 3. The response was strongly augmented by the second immunization and persisted up to at least 45 days. These results suggest that pH-sensitive liposome formula functions as a potential adjuvant of peptide antigens and is useful for the induction of antigen specific CTLsv in vivo. PMID- 11922621 TI - Arachidonate release and eicosanoid generation by group IIE phospholipase A(2). AB - The heparin-binding group II subfamily of secretory phospholipase A(2)s (sPLA(2)s), such as sPLA(2)-IIA and -IID, augments stimulus-induced arachidonic acid (AA) release through the cellular heparan sulfate proteoglycan (HSPG) dependent pathway when transfected into HEK293 cells. Here we show that the closest homolog, sPLA(2)-IIE, also promotes stimulus-induced AA release and prostaglandin (PG) production similar to those elicited by HSPG-dependent sPLA(2)s. Confocal laser microscopic analysis demonstrates the location of sPLA(2)-IIE in cytoplasmic punctate compartments. sPLA(2)-IIE also enhances leukotriene (LT) production and granule exocytosis by RBL-2H3 mastocytoma cells. Expression of sPLA(2)-IIE was highly upregulated in mice injected with lipopolysaccharide (LPS) and in mice with experimental atopic dermatitis. These observations suggest that this enzyme plays a role in the inflammatory process, as proposed for other group II subfamily sPLA(2)s. PMID- 11922622 TI - Isolation of the interacting molecules with GEX-3 by a novel functional screening. AB - To screen for important molecules that interact with a gene of interest in Caenorhabditis elegans (C. elegans), we established a novel functional screening system using the yeast two-hybrid system with the RNA interference technique. Our screening system makes it possible to identify the molecular machinery involved in the function of a gene of interest starting with the cDNA of this gene. As a model case, we examined the molecular machinery involved in the function of GEX 3, an essential factor of tissue morphogenesis. We identified many interacting molecules by yeast two-hybrid screening and could detect some functional interactions using this novel functional screening system. PMID- 11922623 TI - Prediction of the tertiary structure of the beta-secretase zymogen. AB - beta-Secretase, also known as BACE, is a transmembrane aspartyl protease, which generates the N terminus of Alzheimer's disease amyloid beta-peptide. The activity of beta-secretase is the rate-limiting step of brain plaques production in vivo, and hence is a potential target for disease modifying drugs for Alzheimer's disease. To better understand the mechanism of action of beta secretase and help explore novel strategies for drug discovery for Alzheimer's disease, it is important to elucidate the three-dimensional structure of its zymogen. Based on the X-ray structure of the enzyme's protease domain and the X ray structure of pepsinogen, a model of the three-dimensional structure of the beta-secretase zymogen has been constructed. Comparison of the computed structure of pro-BACE with X-ray structures of pepsinogen and progastricsin (two other pro aspartyl proteases) reveals a significant difference in the relationship of the pro-segment to the catalytic aspartates. In both pepsinogen and progastricsin a lysine side-chain in the pro-segment forms a salt bridge to the two catalytic aspartates, occupying the position normally occupied by a catalytic water. In the pro-BACE model there is no salt bridge, and the corresponding residue-a proline does not interact at all with the catalytic residues. These findings can be used to elucidate the recent observations that the pro-domain of beta-secretase does not suppress activity as in a strict zymogen but does appear to facilitate proper folding of an active protease domain. The predicted three-dimensional structure of beta-secretase zymogen and the relevant findings might also provide useful insights for rational design of effective drugs against Alzheimer's disease. PMID- 11922624 TI - Skeletal muscle injury induces hepatocyte growth factor expression in spleen. AB - Hepatocyte growth factor (HGF) is present in skeletal muscle and facilitates skeletal muscle regeneration by activating quiescent satellite cells and stimulating their proliferation. However, possible involvement of HGF from non muscle organs during muscle regeneration is still uncovered. Since liver injury induces HGF expression in distal HGF-producing organs such as lung, kidney and spleen, we examined if this is the case in muscle injury in analogy. In rat femoral muscle, HGF protein levels were elevated within 1 h after muscle injury, with a simultaneous proteolytic activation of HGF protein. Semiquantitative RT PCR analysis revealed an elevation of HGF mRNA expression after muscle injury in the liver and spleen, and also an increase of HGF protein levels in the spleen, suggesting the presence of endocrine HGF-inducing factor(s) during muscle regeneration. Indeed, the sera from the rat with muscle regeneration were capable of inducing HGF mRNA expression when applied to primary cultured spleen cells from intact rats. These results indicated that skeletal muscle injury induces HGF expression in the non-muscle HGF-producing organs, especially in the spleen, and suggested the possible involvement of non-muscle organ-derived HGF in activation/proliferation of satellite cells during muscle regeneration. PMID- 11922625 TI - Identification and characterization of a novel cancer/testis antigen gene CAGE. AB - We applied serological analysis of cDNA expression library technique to identify cancer-associated genes. We screened cDNA expression libraries of human testis and gastric cancer cell lines with sera of patients with gastric cancers. We identified a gene whose expression is testis-specific among normal tissues. We cloned and characterized this novel gene. It contains D-E-A-D box domain and encodes a putative protein of 630 amino acids with possible helicase activity. It showed wide expression in various cancer tissues and cancer cell lines. The corresponding gene was named cancer-associated gene (CAGE). PCR of human x hamster Radiation Hybrids showed localization of CAGE on the human chromosome Xp22. Transient transfection of CAGE showed predominantly nuclear localization. Both Western blot and plaque assay indicated seroreactivity of CAGE protein. We found that demethylation played a role in the activation of CAGE in some cancer cell lines that do not express it. Cell synchronization experiments showed that the expression of CAGE was related with cell cycle. This suggests that CAGE might play a role in cellular proliferation. Because CAGE is expressed in a variety of cancers but not in normal tissues except testis, this gene can be a target of antitumor immunotherapy. PMID- 11922626 TI - Inhibition of hydroxyapatite crystal growth by bone proteoglycans and proteoglycan components. AB - The small leucine-rich proteoglycans (SLRPs) interact with hydroxyapatite (HAP) and have been demonstrated to be important modulators of mineralisation. In the present study we have examined the effect of bone SLRPs, purified bone glycosaminoglycan (GAG) chains and core proteins as well as commercial chondroitin 4-sulphate, chondroitin 6-sulphate and desulphated chondroitin on HAP crystal growth. Seeded HAP growth experiments revealed that addition of bone GAG chains resulted in almost complete inhibition of crystal growth (93%), with addition of core proteins and intact PGs resulting in 55 and 37% inhibition, respectively. In contrast, commercial chondroitin 4-sulphate was significantly less inhibitory compared with the bone SLRPs and components, yielding only a 6% reduction in HAP-induced crystal growth at the same concentration. Significantly, chondroitin 6-sulphate was found to be noninhibitory, whilst desulphated chondroitin was inhibitory to seeded HAP growth. The data indicate that direct adsorption of SLRPs to growth sites and their ability to bind calcium are significant determinants in the inhibitory process. In addition, PG/GAG chemistry and the conformation of the macromolecules in solution have also been shown to be important. This work provides new information regarding the role of bone SLRPs and their components in the regulation of the mineralisation process. PMID- 11922627 TI - Presence of a helix in human CD4 cytoplasmic domain promotes binding to HIV-1 Nef protein. AB - The Nef proteins of simian and human immunodeficiency viruses are known to directly bind and downregulate the CD4 receptor of infected cells. Recent results suggest that residues forming an alpha-helix N-cap in the CD4 cytoplasmic domain play a role in binding of CD4 to human immunodeficiency virus type 1 Nef protein. We determined the dissociation constants between Nef and several CD4 peptides that contain or do not contain the respective alpha-helix N-cap. Further, we compared helical secondary structure content of these CD4 peptide variants by circular dichroism spectroscopy. We conclude that presence of an alpha-helix in CD4 cytoplasmic domain increases CD4 affinity to Nef. In addition, the amino acid sequence of residues forming the helix N-cap influences CD4 affinity to Nef, too. Finally, the structural changes induced in Nef and CD4 upon binding to each other are investigated. PMID- 11922628 TI - AQR1 gene (ORF YNL065w) encodes a plasma membrane transporter of the major facilitator superfamily that confers resistance to short-chain monocarboxylic acids and quinidine in Saccharomyces cerevisiae. AB - We report results on the functional analysis of Saccharomyces cerevisiae ORF YNL065w, predicted to code for a protein belonging to the poorly characterized major facilitator superfamily (MFS) of transporters that are involved in multidrug resistance (MDR). YNL065w is important for a moderate increase of yeast tolerance to ketoconazole and to the cationic dye crystal violet; it protects the cell against short-chain monocarboxylic acids (C(2)-C(6)), but not against highly liposoluble acids such as octanoic acid or the phenoxyacetic-acid herbicides 2,4 D and MCPA; it is also a determinant of resistance to the antiarrhytmic and antimalarial drug quinidine. The encoding ORF was, thus, denominated the AQR1 gene. Results obtained using an AQR1-lacZ fusion indicate that gene expression is very low and it is not stimulated under weak acid stress. The encoded putative transporter was localized in the plasma membrane by fluorescence microscopy observation of the overproduced Aqr1-GFP fusion protein distribution. PMID- 11922629 TI - Secreted CEACAM1 splice variants in rat cell lines and in vivo in rat serum. AB - The widely expressed adhesion receptor CEACAM1 is a member of the carcinoembryonic antigen (CEA) family within the immunoglobulin (Ig) superfamily of glycoproteins. While the expression of transmembrane isoforms has been described in detail, only little is known about soluble isoforms. By RT-PCR characterization of rat pheochromocytoma PC12 and mammary adenocarcinoma MTC cell lines, two novel splice variants, designated CEACAM1-4C1 and CEACAM1-4C2, lacking the transmembrane region, were identified. In addition, we demonstrate the expression of transmembrane CEACAM1-4L and CEACAM1-4S with a truncated cytoplasmic domain. The C-termini of CEACAM1-4C2 and CEACAM1-L are identical, which allowed the specific in vitro and in vivo detection of the soluble CEACAM1 4C2 protein by an antiserum generated against the CEACAM1-L cytoplasmic part. Functionally, soluble CEACAM1 could inhibit CEACAM1-mediated aggregation of CHO cells. In conclusion, our data define a new mechanism for the appearance of functionally active rat CEACAM1 protein in body fluids. PMID- 11922630 TI - Loops within the CD11c I domain critical for specific recognition of fibrinogen. AB - The I domains of CD11 are responsible for the binding of ligands and have a unique structure with 6-7 alpha helices and 6 beta sheets with interconnecting loops. To determine loops recognizing fibrinogen in CD11c I domain, five oligopeptides corresponding to CD11c loops were used to prevent fibrinogen binding to isolated CD11c I domain. The results of the inhibition experiment indicated that all of the loops except the one between E-beta sheet and 6-alpha helix were involved in the binding to fibrinogen. The peptide beta D alpha 5 and alpha 3 alpha 4 showed higher inhibitory activity than others, and the combination of four peptides blocked fibrinogen binding to the I domain completely. These peptides (beta A alpha 1, alpha 3 alpha 4, beta D alpha 5 and beta F alpha 7) could block THP-1 cell binding to fibrinogen coated surface as well. Alanine substitution of amino acids on the I domain such as Y249A and Q201A (which are on the loops beta D-alpha 5 and alpha 3-alpha 4 respectively) abolished fibrinogen binding, while mutation on the loop beta E-alpha 6 (Q273A) had no effect on fibrinogen binding. Taken together, the results from this study suggest that the loops on the top of CD11c I domain such as loop beta A-alpha 1, alpha 3-alpha 4, beta D-alpha 5 and beta F-alpha 7 are involved in fibrinogen binding, and two loops (alpha 3-alpha 4 and beta D-alpha 5) are more important than others for the recognition of fibrinogen. PMID- 11922631 TI - Osteoblastic differentiation induced by transcription factor decoy against estrogen receptor alpha gene. AB - Osteoblasts have been shown to express both isoforms of estrogen receptor (ER alpha and ER beta). As a tool for the study of endogenous regulation of these genes the decoy strategy was employed. Human MG-63 osteoblast-like cells were transfected with a DNA decoy molecule containing a putative negative cis-element (DNA-102) located in the C distal promoter of ER alpha gene. Using real-time quantitative RT-PCR, we found that the DNA-102, but not scrambled DNA, produced a 36-fold increase in the level of total ER alpha mRNA and a 12-fold increase in the level of mRNA for the F isoform that is transcribed from the upstream F promoter, which is predominantly used in osteoblasts. This effect appears to be controlled by estrogen since 17-beta-estradiol downregulated the mRNA increase. Notably, the same decoy was able to induce a 6-fold increase in ER beta mRNA transcription, indicating the coregulation of the ER alpha and ER beta expression. An increase in OPN but not in BMP4 expression was also observed. In addition, in decoy-treated cells, the cell growth decreased together with an increase in alkaline phosphatase activity. These findings indicated that DNA-102 decoy was able to induce a more differentiated osteoblastic phenotype. The augmentation of ER alpha and ER beta expression by the decoy approach may offer a further possibility for patient response to estrogenic therapy in the treatment of diseases related to estrogen deficiency. PMID- 11922632 TI - Identification and characterization of a novel hypertonicity-responsive element in the human aquaporin-1 gene. AB - The AQP1 gene is transcriptionally upregulated in response to hypertonicity. However, the molecular mechanism in hypertonicity-induced transcription in the AQP1 gene has been poorly understood. Here we report a novel hypertonicity response element (HRE) in the AQP1 gene. A critical cis-acting element to hypertonicity in the AQP1 promoter is located at -54 to -46 and the change from GCTCCCCCC to GCTTTCCCC completely abolished the hypertonic induction and osmotic response, thus implicating the importance of the first CC sequence in the region. Two prominent DNA-protein complexes were observed electrophoretic mobility shift assay (EMSA), and the band intensities in nuclear extracts from osmotically stressed cells were much higher than that of isotonic nuclear extracts. The EMSA supershift assay with anti-Sp1 antibody showed that two retarded bands did not bind to anti-Sp1 antibody, suggesting that the DNA binding proteins bound to the HRE are not Sp1 family proteins. These data suggest that the transcription of the AQP1 by hypertonicity in renal cells is upregulated by the interaction with putative DNA binding proteins to a novel HRE located at -54 to -46 in the AQP1 gene. PMID- 11922633 TI - Genetic variants of the receptors for thromboxane A2 and IL-4 in atopic dermatitis. AB - Thromboxane A2 (TXA2) is an arachidonate metabolite which is considered to relate to chronic inflammation in atopic diseases characterized by elevated immunoglobulin E productivity. The elevation of immunoglobulin E levels involves many molecules including interleukin-4 (IL-4) and interleukin-4 receptor alpha chain (IL-4R alpha). To assess whether genetic variants of TXA2 receptor, IL-4 and IL-4R alpha genes relate to the elevation of serum immunoglobulin E levels in patients with atopic dermatitis (AD), we conducted an association study of genetic polymorphisms of TXA2 receptor (795C/T), IL-4 (-589C/T), and IL-4R alpha (Ile50Val) in a Japanese population (n = 789). The TXA2 receptor 795TT genotype strongly related to AD with high serum immunoglobulin E concentrations. AD patients with both TXA2 receptor 795TT genotype and the IL-4R alpha Ile50/Ile50 genotype showed the greatest immunoglobulin E concentrations. These results suggest TXA2 receptor polymorphism strongly interacts with IL-4R alpha polymorphism as a major determinant of high serum immunoglobulin E levels in AD. PMID- 11922634 TI - Increased plasma HB-EGF associated with obesity and coronary artery disease. AB - The mechanism by which the obese subjects are more associated with vascular disease remains unclear. We reported that the adipose tissues produce and secrete many bioactive molecules, conceptualized as adipocytokines. Heparin-binding epidermal growth factor (EGF)-like growth factor (HB-EGF), produced locally by vascular macrophages and smooth muscle cells, has been suggested to induce the migration and proliferation of vascular smooth muscle cells. The current study reveals that (1) HB-EGF mRNA is abundantly expressed in human adipose tissue, (2) HB-EGF mRNA increases in the fat tissues of obese mice, (3) plasma HB-EGF levels increase in parallel with fat accumulation in human, and (4) the subjects with coronary artery disease have higher plasma HB-EGF levels, associated with fat accumulation. These results suggest that increased plasma HB-EGF derived from the accumulated fat contributes to the higher incidence of vascular disease in obesity, proposing HB-EGF as an adipocytokine directly linking adipovascular axis. PMID- 11922635 TI - Definition of new targets in cardiovascular prevention from young into old age. PMID- 11922636 TI - A prediction rule for left ventricular dilatation post-MI? PMID- 11922637 TI - Coronary bypass surgery vs coronary angioplasty: a look to the past for new insights into the future. PMID- 11922638 TI - Atrial fibrillation--a curable condition? PMID- 11922639 TI - Enterovirus infection of the heart--a causal or contributory factor in chronic rheumatic heart disease? PMID- 11922640 TI - Dobutamine stress echocardiography: the long and short of it. PMID- 11922641 TI - A personal ultrasound imager (ultrasound stethoscope). A revolution in the physical cardiac diagnosis! PMID- 11922642 TI - Combined effects of systolic blood pressure and serum cholesterol on cardiovascular mortality in young (<55 years) men and women. AB - AIMS: To evaluate the combined effects of the two most frequent modifiable risk factors, systolic blood pressure and serum cholesterol, on cardiovascular and coronary mortality, in a large French population aged 18 to 55 years. METHODS AND RESULTS: We studied 108,879 men (mean age 39.1+/-9.4 years) and 84,931 women (mean age 37.3+/-10.0 years) who had a health check-up at the IPC Center between 1978 and 1988. Mortality data for a mean period of 13 years were analysed. Systolic blood pressure and cholesterol levels were classified according to the cut-points proposed by international guidelines. In men, the prevalence of high cholesterol was more than twice as high in hypertensives as in normotensives; in women, it was more than three times higher. The combination of these two risk factors has additive effects on cardiovascular disease and coronary heart disease risk. In men, a borderline elevation of both systolic blood pressure (130-139 mmHg) and cholesterol (200-239 mg x dl(-1)) leads to a three- to four-fold increase in cardiovascular disease risk. Men with systolic blood pressure >or=160 mmHg represent a small percentage (about 5%) who have a 10-fold increase in cardiovascular disease and coronary heart disease risk, especially when high cholesterol is present. In women of the same age, similar trends were observed, but the results were less significant, probably due to the low cardiovascular disease mortality rates. CONCLUSIONS: In conclusion, in French subjects under 55 years of age, a combination of high systolic blood pressure and high serum cholesterol dramatically increased cardiovascular disease and coronary heart disease risk, especially in men. A more aggressive public health policy is needed to prevent the development of risk factors in younger subjects. PMID- 11922643 TI - Prediction of 6 months left ventricular dilatation after myocardial infarction in relation to cardiac morbidity and mortality. Application of a new dilatation model to GISSI-3 data. AB - AIMS: To predict the long-term left ventricular volume index early after myocardial infarction and to investigate the relationship between long-term left ventricular dilatation risk and clinical outcome. METHODS AND RESULTS: By applying a previously developed dilatation model, we predicted the 6-month left ventricular volume index early after myocardial infarction (median 9 days) in 13,679 GISSI-3 patients, to identify patients at high risk of long-term left ventricular dilatation. The left ventricular systolic and diastolic volume indexes at 6 months were predicted with r=0.72 and r=0.68, respectively, in the subgroup of patients in whom a pre-discharge echo was available (n=7842). Patients predicted to be at risk for long-term left ventricular dilatation had an increased risk of mortality (RR 1.87, 95% CI: 1.48 to 2.36) and heart failure at 6 months (RR 2.59, 95% CI:2.04 to 3.28), but no increased risk of reinfarction at 6 months (RR 1.12, 95% CI: 0.87 to 1.45) or of angina pectoris (RR 1.07, 95% CI: 0.95 to 1.20). CONCLUSION: Our prediction of long-term left ventricular dilatation, obtained by applying our new dilatation model in over 13,000 GISSI-3 patients, correlated well with mortality and heart failure after myocardial infarction. Therefore, our new dilatation model may contribute to more efficient risk stratification early after myocardial infarction. PMID- 11922644 TI - Coronary artery bypass graft surgery and percutaneous transluminal coronary angioplasty. Twenty-year clinical outcome. AB - AIMS: The purpose of this study is to compare the long-term outcome (up to 20 years) of coronary artery bypass surgery (CABG) with percutaneous transluminal coronary angioplasty (PTCA) in a consecutive patient series at a single centre. Survival is similar after CABG and PTCA up to 8 years follow-up in patients with multivessel disease, with a reduced need for repeat revascularization after CABG. As coronary artery disease is a lifetime disease, longer-term follow-up of these revascularization therapies is necessary to help clinical decision-making. METHODS AND RESULTS: The CABG study population consisted of the first 1041 consecutive patients who underwent a first elective coronary bypass surgery between 1970 and 1980. The PTCA study population consisted of 702 consecutive patients who underwent a first elective coronary angioplasty procedure between 1980 and 1985. Mortality and subsequent revascularization up to 20 years were captured. Survival rates were adjusted using proportional hazards methods to account for baseline differences. RESULTS: The unadjusted survival rates were 92%, 77%, 57% and 49% after CABG at respectively, 5-, 10-, 15- and 17 years and 91%, 80%, 64% and 59% after PTCA. In the multivessel disease subgroup, survival was similar with a benefit apparent after CABG in the first 8 years of follow-up. The therapy chosen, CABG or PTCA, was a univariate predictor of mortality in favour of PTCA (RR: 1.28; 95% CI: 1.10-1.49), but after correction for baseline characteristics, the relative risk of mortality for CABG vs PTCA was comparable (RR: 1.03; 95% CI: 0.87-1.24). The adjusted survival curves in the subgroup of diabetic elderly patients with multivessel disease were similar after the tenth year with only a slightly better survival in the CABG population in the first 10 years. Repeat intervention was more frequently required after PTCA during the first 8 years, but after this time more frequently in the CABG group. CONCLUSION: When comparing CABG and PTCA it can be concluded that both strategies are equally effective in terms of 20-year survival. In particular, after more than 10 years all differences tend to disappear. While repeat intervention was significantly higher in the first year after PTCA, after 7-8 years, reintervention was greater in patients who had initial CABG. PMID- 11922645 TI - Clinical characteristics and outcome of patients with early (<2 h), intermediate (2-4 h) and late (>4 h) presentation treated by primary coronary angioplasty or thrombolytic therapy for acute myocardial infarction. AB - AIMS: We examined the clinical characteristics and outcome of patients with early (<2 h), intermediate (2-4 h) and late (>4 h) presentation treated by primary angioplasty or thrombolytic therapy for acute myocardial infarction. METHODS AND RESULTS: We studied 2635 patients enrolled in 10 randomized trials of primary angioplasty (n=1302) vs thrombolytic therapy (n=1333) in acute myocardial infarction, and baseline characteristics of the two groups were comparable. Increase in presentation delay is associated with older age, female gender, diabetes and an increased heart rate. We classified the patients according to the time delay from symptom onset to presentation into three categories: early presentation (<2 h), intermediate presentation (2-4 h), and late presentation (>or=4 h). At 30 days the combined rate of death, non-fatal reinfarction and stroke in patients presenting early was 5.8% in the angioplasty group vs 12.5% in the thrombolysis group, in patients with intermediate presentation, 8.6% vs 14.2%, respectively, and in patients presenting late 7.7% vs 19.4%, respectively. With increasing time from symptom onset to presentation, all major adverse cardiac event rates show a trend to a larger increase in the thrombolysis group compared to the angioplasty group, both at 30 days and at 6 months after the acute event. CONCLUSIONS: Major adverse cardiac event rates are lower after angioplasty compared to thrombolysis, irrespective of time to presentation. With increasing time to presentation major adverse cardiac event rates increase after thrombolysis but appear to remain relatively stable after angioplasty. PMID- 11922646 TI - Efficacy of an additional MAZE procedure using cooled-tip radiofrequency ablation in patients with chronic atrial fibrillation and mitral valve disease. A randomized, prospective trial. AB - AIMS: This study is the first prospective randomized trial evaluating the efficacy of an antiarrhythmic surgical procedure in patients with chronic atrial fibrillation undergoing mitral valve replacement. METHODS AND RESULTS: Thirty consecutive patients with chronic atrial fibrillation undergoing mitral valve replacement were randomized for an additional modified MAZE-operation using intra operatively cooled-tip radiofrequency ablation (group A) or mitral valve replacement alone (group B). Biatrial contraction was studied and functional capacity was evaluated in spiro-ergometry 6 months after surgery. Thirty-day mortality was 0% in both groups. After 12 months, sinus rhythm was reinstituted significantly more often in patients of group A (cumulative rate of sinus rhythm 0.800) compared to patients in group B (0.267) (P<0.01). 66.7% of patients in sinus rhythm of group A had documented biatrial contraction. Electrocardioversion showed long-term success in only 17% of patients in group A and 0% in group B. Maximal aerobic uptake at the 6-month spiro-ergometry revealed no significant difference (9.3 vs 8.5 ml x min(-1) kg(-1), P=0.530). CONCLUSIONS: A modified MAZE operation using cooled-tip radiofrequency ablation can be safely combined with mitral valve surgery and is highly effective in restoring sinus rhythm. Biatrial contraction is found in 66.7% of patients with sinus rhythm undergoing mitral valve replacement plus the MAZE operation. PMID- 11922647 TI - Enterovirus replication in valvular tissue from patients with chronic rheumatic heart disease. AB - AIMS: To investigate the involvement of enterovirus infection in chronic, rheumatic heart disease. METHODS AND RESULTS: Formalin-fixed, paraffin-embedded, surgical samples of valve tissue were examined for the presence of enteroviral RNA and virus capsid protein VP1 by in situ hybridization and immunostaining. Of 53 cases, 33 were patients with chronic rheumatic heart disease and 20 had Marfan's syndrome or degenerative valve disease. Enterovirus RNA was detected in 8 (24.2%) of 33 patients with chronic rheumatic heart disease by in situ hybridization using strand-specific oligonucleotide probes, complementary to conserved sequences in enterovirus genomic (positive strand) RNA. The replication template (negative strand) RNA also was found in seven of these eight cases. The viral capsid protein VP1 was detected in 16 (48.5%) of 33 patients with chronic rheumatic heart disease by immunohistochemistry and correlated with viral RNA detection. Virus was localized generally to valvular tissue. Neither viral RNA nor capsid protein VP1 were found in valvular tissue from any of the 20 comparison cases. CONCLUSIONS: This is the first demonstration of detection and localization of both enterovirus RNA and capsid protein in chronic rheumatic heart disease. The presence of negative strand RNA and VP1 indicates enteroviral RNA replication and protein synthesis and suggests an aetiological role of enterovirus in the pathogenesis of chronic rheumatic heart disease. PMID- 11922648 TI - Determinants of costs and resource utilization associated with open heart surgery. AB - AIMS: This study sought to determine the patient- and the therapy-related determinants of in-hospital costs for patients undergoing heart surgery at the University Hospital in Zurich. METHODS AND RESULTS: We performed a retrospective analysis of all adult cardiac surgical patients from the canton St. Gallen who were covered by a fixed fee arrangement (29,500 Swiss francs (19,470 Euro)) and referred to our institution during 1998. A total of 201 patients (143 (71%) male) with basic insurance were hospitalized in 1998 under the fixed fee arrangement. The mean age of the patients was 61.4 years (95% confidence intervals (CI): 60; 63). With the help of univariate analysis, the following pre-operative characteristics were found to be significantly associated with cost: age (P<0.001), pre-operative cardiac diagnosis (coronary vs valvular heart disease) (P<0.001) and EuroSCORE (P<0.0001). A significant correlation was also found between intra-operative variables and costs (P<0.0001) as well as between postoperative variables and costs (P<0.0001). A linear regression model based on EuroSCORE, operation time and postoperative infection status is able to predict costs for patients (all P -values <0.0001, except for P<0.05 for operation time, R(2)=0.565). CONCLUSIONS: These results suggest that both pre-operative (patient related) and intra-operative (therapy- and patient-related) variables are predictors of costs in cardiac surgical patients. PMID- 11922649 TI - A comparison of wall motion analysis and systolic left ventricular long axis function during dobutamine stress echocardiography. AB - AIMS: To compare long-axis function and wall motion analysis for the detection of significant coronary artery stenoses in patients with single and multivessel disease. METHODS AND RESULTS: We performed dobutamine stress echocardiography in 67 subjects, 14 with normal coronary anatomy, and 53 with significant coronary disease. A blunted increase in mean long-axis shortening of <0.25 cm was the best discriminator for coronary artery disease (sensitivity 85%, specificity 81%). Using this threshold, long axis function gave a sensitivity of 88% and specificity 89% for the detection of coronary artery disease in patients with normal resting wall motion while wall motion abnormality analysis had a sensitivity 73% and specificity 94%. Of 26 patients with a resting wall motion abnormality, 14 (54%) had multivessel disease. Long axis function detected multivessel disease in 12 of these (sensitivity 86%) compared with nine (sensitivity 64%) for wall motion analysis. CONCLUSION: Long axis function provides a promising, quantitative adjunct to wall motion analysis for the detection of coronary ischaemia using dobutamine stress echocardiography in patients with single and multivessel disease and with resting wall motion abnormalities. PMID- 11922650 TI - Interference of two common European digital cellular phones with implantable cardioverter-defibrillators. PMID- 11922651 TI - Maintainance of sinus rhythm after electrical cardioversion of persistent atrial fibrillation. PMID- 11922655 TI - Building a bridge: engineering spinal cord repair. AB - Injuries to the spinal cord that result in disruption of axonal continuity have devastating consequences for injured patients. Current therapies that use biologically active agents to promote neuronal survival and/or growth have had modest success in allowing injured neurons to regrow through the area of the lesion. Strategies for successful regeneration will require an engineering approach. We propose the design of cell-free grafts of biocompatible materials to build a bridge across the injured area through which axons can regenerate. There are three critical regions of this bridge: the on-ramp, the surface of the bridge itself, and the off-ramp. Each of these regions has specific design requirements, which, if met, can promote regeneration of axons in the injured spinal cord. These requirements, and proposed solutions, are discussed. PMID- 11922656 TI - Changes in astroglial GLT-1 expression after neural transplantation or stab wounds. AB - Uncontrolled release of glutamate from damaged brain initiates events that result in excitotoxic neuronal death. Glutamate uptake by specialized astroglial transporters is essential for control of extracellular glutamate levels. Many studies have demonstrated a reduction in astrocytic GLT-1 expression after different forms of injury. Because extensive neuronal death does not occur after direct cortical stab wounds and viable developing neurons populate fetal CNS grafts, we hypothesized that reactive astroglia associated with these procedures might maintain or up-regulate GLT-1. We examined the temporal and spatial distribution of GLT-1, GFAP and nestin proteins by confocal double-label immunohistochemistry combined with a new methodology in which precise brain areas are microdissected and analyzed for protein content by immunoaffinity chromatography. In stab wounds, GLT-1 protein content did not change compared to normal cortex, as determined by direct protein measurements; GLT-1 colocalized with nestin- and GFAP(+) astroglia adjacent to the lesion. In contrast, host reactive astroglia adjacent to grafts significantly upregulated GLT-1 by 3 days postoperative. The GFAP protein analysis suggests that increased GLT-1 is not the result of greater numbers of activated astroglia around grafts, but that developing graft tissue influences adjacent host astroglia to upregulate GLT-1. GLT-1 protein within grafts was rapidly accelerated to mature levels by just three days, and was expressed by the nestin(+) cell population. These data, which demonstrate immunoexpression of GLT-1 protein combined with a new method for protein measurement in situ indicate that, in contrast to other injury models, astroglial GLT-1 is upregulated or maintained following invasive CNS procedures. (c)2002 Elsevier Science (USA). PMID- 11922657 TI - Protective effect of parvalbumin on excitotoxic motor neuron death. AB - The mechanism responsible for the selective vulnerability of motor neurons in amyotrophic lateral sclerosis (ALS) is poorly understood. Several lines of evidence indicate that susceptibility of motor neurons to Ca(2+) overload induced by excitotoxic stimuli is involved. In this study, we investigated whether the high density of Ca(2+)-permeable AMPA receptors on motor neurons gives rise to higher Ca(2+) transients in motor neurons compared to dorsal horn neurons. Dorsal horn neurons were chosen as controls as these cells do not degenerate in ALS. In cultured spinal motor neurons, the rise of the cytosolic Ca(2+) concentration induced by kainic acid (KA) and mediated by the AMPA receptor was almost twice as high as in spinal neurons from the dorsal horn. Furthermore, we investigated whether increasing the motor neuron's cytosolic Ca(2+)-buffering capacity protects them from excitotoxic death. To obtain motor neurons with increased Ca(2+) buffering capacity, we generated transgenic mice overexpressing parvalbumin (PV). These mice have no apparent phenotype. PV overexpression was present in the central nervous system, kidney, thymus, and spleen. Motor neurons from these transgenic mice expressed PV in culture and were partially protected from KA-induced death as compared to those isolated from nontransgenic littermates. PV overexpression also attenuated KA-induced Ca(2+) transients, but not those induced by depolarization. We conclude that the high density of Ca(2+) permeable AMPA receptors on the motor neuron's surface results in high Ca(2+) transients upon stimulation and that the low cytosolic Ca(2+)-buffering capacity of motor neurons may contribute to the selective vulnerability of these cells in ALS. Overexpression of a high-affinity Ca(2+) buffer such as PV protects the motor neuron from excitotoxicity and this protective effect depends upon the mode of Ca(2+) entry into the cell. PMID- 11922658 TI - Transplanted clonal neural stem-like cells respond to remote photic stimulation following incorporation within the suprachiasmatic nucleus. AB - Multipotent neural stem-like cells (NSCs) obtained from one brain region and transplanted to another region appear to differentiate into neuronal and glial phenotypes indigenous to the implantation site. Whether these donor-derived cells are appropriately integrated remains unanswered. In order to test this possibility, we exploited the suprachiasmatic nucleus (SCN) of the hypothalamus, site of a known circadian clock, as a novel engraftment target. When a clone of NSCs initially derived from neonatal mouse cerebellum was transplanted into mouse embryos, the cells incorporated within the SCN over a narrow gestational window that corresponded to the conclusion of SCN neurogenesis. Immunocytochemical staining suggested that donor-derived cells in the SCN synthesized a peptide neurotransmitter (arginine vasopressin) characteristic of SCN neurons. Donor derived SCN cells reacted to light pulses by expressing immunoreactive c-Fos protein in a pattern that is appropriate for native SCN cells. This region specific and physiologically appropriate response to the natural stimulation of a remote sensory input implies that donor-derived and endogenous cells formed true SCN chimeras, suggesting that exogenous NSCs engrafted to ectopic locations can integrate in a meaningful fashion. PMID- 11922659 TI - Positive effect of transplantation of hNT neurons (NTera 2/D1 cell-line) in a model of familial amyotrophic lateral sclerosis. AB - Transplantation of hNT Neurons derived from the human teratocarcinoma cell-line (NTera2/D1) has been shown to ameliorate motor dysfunction in a number of injury or disease models in which the deficits are fairly localized. However, these cells have not been used before in a model with more extensive neurodegeneration. The aim of this study is to determine the effects of hNT Neuron transplants on motor neuron function in a mouse model of familial amyotrophic lateral sclerosis (FALS) in which there is a substitution of Alanine for Glycine at position 93 of the human SOD1 gene (G93A). Amyotrophic lateral sclerosis is a fatal degenerative motor neuron disease affecting the spinal cord, brainstem, and cortex. This disease clinically manifests as progressive muscular weakness and atrophy, leading to paralysis and death within 3-5 years of diagnosis. The FALS represents 10-13% of all cases. A range of behavioral tests was used to examine spontaneous locomotor activity, coordination, and muscle strength of mice. Long-term (10-11 weeks) transplantation of hNT Neurons into the L(4)-L(5) segments of the ventral horn spinal cord of FALS(G93A) mice at 7 weeks of age (before onset of overt behavioral symptoms of disease) delayed the onset of motor dysfunction for at least 3 weeks. The average lifespan of the transplanted mice was 128 days compared to 106 days for media-injected group. The last mouse in the hNT Neuron transplanted group was euthanized at 135 days of age when it display partial paralysis of the hindlimbs. Immunohistochemical analysis of the implanted spinal cords demonstrated the survival of grafted hNT Neurons and showed many healthy appearing motor neurons near the implant site. These results suggest that hNT Neuron transplantation may be a promising therapeutic strategy for ALS. PMID- 11922660 TI - Intrapericardial procaine affects volume expansion-induced fos immunoreactivity in unanesthetized rats. AB - Acute volume expansion is associated with a specific pattern of Fos expression and the goal of the present study was to evaluate the contribution of cardiac receptors to this response. Adult male rats were instrumented with pericardial catheters introduced at the level of the thymus. Rats were also catheterized for measuring blood pressure, heart rate, central venous pressure, and intravenous infusion. Each rat received a 200-microl intrapericardial (i.p.c) injection of 2% procaine or 0.9% NaCl. Rats were then volume expanded with isotonic saline (10% body weight in 10 min) or given a control infusion (0.01 ml/min for 10 min). Ninety minutes after the start of the infusion, the rats were anesthetized and perfused transcardially. Their brains were sectioned and processed for Fos, dopamine-beta-hydroxylase, and oxytocin immunocytochemistry. Volume expansion plus i.p.c. saline produced a significant increase in Fos expression in the nucleus of the solitary tract, the ventrolateral medulla, the area postrema, the locus coeruleus, the paraventricular nucleus of the hypothalamus, the perinuclear zone of the supraoptic nucleus, and oxytocin neurons in the supraoptic nucleus. The i.p.c. procaine significantly blocked Fos expression produced by the volume expansion in the all of the regions examined except for the area postrema and the SON oxytocin neurons. PMID- 11922661 TI - TNFalpha transport across the blood-brain barrier is abolished in receptor knockout mice. AB - The presence of transport systems at the blood-brain barrier (BBB) enables some cytokines in blood to reach specific targets in the brain and spinal cord. The "transporters" function in a way different from conventional receptors, in that cytokines are chaperoned from blood to the CNS rather than being degraded in the specialized endothelial cells composing the BBB. Here we present the first study to determine whether the transporter for tumor necrosis factor-alpha (TNFalpha) is identical to its receptors. Three types of TNFalpha receptor knockout mice were used, and the influx of (125)I-TNFalpha from blood to brain and blood to spinal cord was measured. In either p55 or p75 receptor knockout mice, the influx of (125)I-TNFalpha was significantly, but not completely, decreased in spinal cord, whereas the decrease in brain was not statistically significant. This indicates that both receptors are partially involved in the transport of TNFalpha across the BBB but that neither receptor is the sole transporter. By contrast, in double knockout mice lacking both p55 and p75 receptors, the entry of (125)I TNFalpha into brain and spinal cord was completely abolished. Therefore, both receptors are necessary for transporting TNFalpha across the BBB. The results clearly demonstrate that the transport of TNFalpha across the BBB is a complicated process involving additive or even synergistic activities of both receptors, thus differing from typical ligand-receptor binding and downstream signal transduction. PMID- 11922662 TI - Spontaneous limbic seizures after intrahippocampal infusion of brain-derived neurotrophic factor. AB - The results of several studies have contributed to the hypothesis that BDNF promotes seizure activity, particularly in adult hippocampus. To test this hypothesis, BDNF, vehicle (phosphate-buffered saline, PBS), or albumin was infused directly into the hippocampus for 2 weeks using osmotic minipumps. Rats were examined behaviorally, electrophysiologically, and anatomically. An additional group was tested for sensitivity to the convulsant pilocarpine. Spontaneous behavioral seizures were observed in BDNF-infused rats (8/32; 25%) but not in controls (0/20; 0%). In a subset of six animals (three BDNF, three albumin), blind electrophysiological analysis of scalp recordings contralateral to the infused hippocampus demonstrated abnormalities in all BDNF rats; but not controls. Neuronal loss in BDNF-treated rats was not detected relative to PBS- or albumin-treated animals, but immunocytochemical markers showed a pattern of expression in BDNF-treated rats that was similar to rats with experimentally induced seizures. Thus, BDNF-infused rats had increased expression of NPY in hilar neurons of the dentate gyrus relative to control rats. NPY and BDNF expression was increased in the mossy fiber axons of dentate gyrus granule cells relative to controls. The increase in NPY and BDNF expression in BDNF-treated rats was bilateral and occurred throughout the septotemporal axis of the hippocampus. Mossy fiber sprouting occurred in five BDNF-treated rats but no controls. In another group of infused rats that was tested for seizure sensitivity to the convulsant pilocarpine, BDNF-infused rats had a shorter latency to status epilepticus than PBS-infused rats. In addition, the progression from normal behavior to severe seizures was faster in BDNF-treated rats. These data support the hypothesis that intrahippocampal BDNF infusion can facilitate, and potentially initiate, seizure activity in adult hippocampus. PMID- 11922663 TI - Postnatal development of zinc-rich terminal fields in the brain of the rat. AB - The appearance and distribution of zinc-rich terminal fields in the rat forebrain was analyzed at 12 stages of postnatal development using the selenium method. Zinc stain was detected in neonates in piriform, cingulate, and motor cortices, septal area, and hippocampal formation. In the neocortex, a laminar pattern appeared progressively following an inside-out gradient: layer VI at postnatal day 0 (P0), layer V at P1, layers Va and Vb at P5, layer II-III at P9, and layer IV at P12. In the hippocampal formation the layered pattern in the dentate molecular layer appeared at P1-P3, and in the hilus and mossy fibers the stain was observed at P5. Patches in the caudate-putamen were sharply delimited at P1 P3. At these ages, staining was observed in the amygdaloid complex. In the thalamic and hypothalamic nuclei, stain appeared at P5-P7. Thus, a general increase in vesicular zinc over different telencephalic areas was determined until P15-P21, which was followed by a slight decrease thereafter (at P41). The increased stain in zinc-rich terminal fields is consistent with the development of telencephalic circuits. The rise in zinc might be relevant for the establishment and maturation of these circuits. On the other hand, the decrease in staining for zinc at later stages might be due to methodological problems but it might also reflect pruning of supernumerary connections and programmed cell death affecting zinc-rich circuits. PMID- 11922664 TI - Astrocyte delivery of glial cell line-derived neurotrophic factor in a mouse model of Parkinson's disease. AB - Primary astrocytes were genetically modified ex vivo to express recombinant glial cell line-derived neurotrophic factor (GDNF) and subsequently were tested for their ability to provide neuroprotection to dopaminergic neurons in a 6 hydroxydopamine (6-OHDA) mouse model of Parkinson's disease. A replication defective retrovirus was constructed, which contained the rat GDNF sequence and a sequence encoding a beta-galactosidase (beta-gal)/neomycin phosphotransferase fusion protein, linked via an internal ribosomal entry site. Murine astrocytes transduced with this vector secreted GDNF into the culture media at the rate of 115 +/- 34 pg/24 h/10(5) cells and expressed cytoplasmic beta-gal, whereas control nontransduced astrocytes were negative for GDNF production and cytoplasmic beta-gal expression. Mice that received implants of GDNF-producing astrocytes into the striatum or nigra displayed elevated levels of GDNF compared to mice that received control nontransduced astrocytes. In addition, tissue content of GDNF was increased bilaterally and in brain regions both proximal and distal to the graft, even though astrocyte migration away from the graft site did not occur. Importantly, GDNF-producing astrocytes provided marked neuroprotection of nigral dopaminergic perikarya, and partial protection of striatal dopaminergic fibers, when implanted into the midbrain 6 days prior to a retrograde 6-OHDA lesion, as assessed by tyrosine hydroxylase immunohistochemistry. Similarly, GDNF producing astrocytes prevented the acquisition of amphetamine-induced rotational behavior in 6-OHDA-treated mice and completely prevented dopamine depletion within the substantia nigra, as assessed by high-performance liquid chromatography. These results indicate that continuous exposure to low levels of GDNF provided by transgenic astrocytes provides marked neuroprotection of nigral dopaminergic neurons. (c)2002 Elsevier Science (USA). PMID- 11922665 TI - Excitatory amino acids differentially regulate the expression of GDNF, neurturin, and their receptors in the adult rat striatum. AB - Glial cell line-derived neurotrophic factor (GDNF) family ligands are important regulators of neuronal development and maintenance of the connectivity in the basal ganglia and show neuroprotective activities in several paradigms of brain injury. The mRNAs of two members of this family, GDNF and neurturin, and also their receptors have been detected in the basal ganglia. In the present work, we analyzed the time course changes in the expression of these neurotrophic factors and receptors in the adult rat striatum, induced by quinolinate or kainate excitotoxicity. Our results show that stimulation of NMDA or non-NMDA receptors induced different effects on the mRNA levels analyzed. Expression of GDNF and its preferred receptor, GDNF family receptor-alpha1 (GFRalpha1), was transiently up regulated by quinolinate and kainate, but with differing intensity and temporal pattern. Immunohistochemical analysis showed that, although GDNF and GFRalpha1 were initially localized in neurons, excitotoxicity induced the expression of these proteins in astrocyte-like cells. Neurturin mRNA levels were only up regulated after quinolinate injection, whereas quinolinate or kainate injection did not modify GFRalpha2 mRNA. The mRNA for the common receptor, c-Ret, was up regulated by both agonists with similar temporal pattern but with differing intensity. Immunohistochemical analysis showed that c-Ret protein was located on neurons. These changes in mRNA levels and protein localization of GDNF family components could reflect an endogenous trophic response of striatal cells to different excitotoxic insults. PMID- 11922666 TI - Amyloid beta-peptide decreases neuronal glucose uptake despite causing increase in GLUT3 mRNA transcription and GLUT3 translocation to the plasma membrane. AB - Amyloid beta-peptide (Abeta) has been shown to impair glucose uptake in cultured hippocampal neurons and shortens their survival time. Abeta appears to inhibit neuronal glucose uptake by activating Gs-coupled receptors and the cAMP-PKA system. In this study, Abeta inhibition of neuronal glucose uptake was studied by assaying translocation of glucose transporter isoform GLUT3, transcription of GLUT3 mRNA, and fusion of GLUT3-containing vesicles with the plasma membrane. Cultured hippocampal neurons exposed to 10 microM Abeta25-35 or Abeta1-40 for 3 or 24 h showed a significant decrease in glucose uptake. To assess the regulatory role of Abeta on neuronal glucose uptake, translocation of GLUT3 from the cytosol to the plasma membrane was studied by the plasma membrane lawn assay and transcription of GLUT3 mRNA by in situ hybridization. In spite of a decrease in glucose uptake, Abeta25-35 and Abeta1-40 (10 microM) markedly promoted GLUT3 translocation to the plasma membrane by 30 min. Abeta25-35 also up-regulated transcription of GLUT3 mRNA by 12 h. High extracellular K(+) increased immunolabeling of the exofacial (i.e., extracellular) epitope of GLUT3 at the plasma membrane and Abeta25-35 inhibited this increase. Based on these data we propose that Abeta increases translocation of GLUT3-containing vesicles, but inhibits their fusion with the plasma membrane. PMID- 11922667 TI - A novel method of labeling and characterizing migrating neurons in the developing central nervous system. AB - Neuronal migration, a discrete event in the developing nervous system, is currently being intensively investigated using a variety of anatomical and molecular approaches. Using 4-chloromethyl benzoyl amino tetramethyl rhodamine (CMTMR) coated particles, we describe here a novel and efficient method of tracer labeling to investigate cell migration in embryonic and postnatal brain. Further, we demonstrate that application of CMTMR facilitates the labeling of a large number of migrating cells and enables the characterization of their phenotypes with immunohistochemical and in situ hybridization techniques. We also illustrate that CMTMR labeling is ideally suited for time-lapse imaging of the behavior and dynamics of migrating cells. PMID- 11922668 TI - In vivo dissection of the Tat translocation pathway in Escherichia coli. AB - The bacterial Tat pathway is capable of exporting folded proteins carrying a special twin arginine (RR) signal peptide. By using two in vivo reporter proteins, we assessed factors that affect Tat pathway transport. We observed that, like the intact RR signal peptide, those with a KR or RK substitution were still capable of mediating the translocation of the folded green fluorescent protein (GFP). However, the translocation efficiency decreased in the order of RR>KR>RK. The KK motif was unable to mediate GFP translocation. The translocation of the RR-GFP fusion required TatA, TatB and TatC proteins. By exploiting the periplasmic bactericidal property of colicin V (ColV), we constructed a translocation-suicide probe, RR-ColV. The translocation of RR-ColV fully inhibited the growth of wild-type Escherichia coli and those of the DeltatatD and DeltatatE mutants. In contrast, the deletion of the tatC gene blocked RR-ColV in the cytoplasm and this strain exhibited a normal growth phenotype. Interestingly, the growth of DeltatatA and tatB mutants was inhibited partially by RR-ColV. Moreover, KR, RK and KK motifs were capable of mediating the ColV translocation with a decreasing RR=KR>RK>KK efficiency. In addition to TatE and TatC proteins, either TatA or TatB was sufficient for the translocation of RR-ColV or KR-ColV. In contrast, TatA plus the conserved N-terminal domain of TatB were required to mediate the killing effect of ColV fused to the less-efficient RK signal peptide. Taken together, these results suggest that a fully efficient Tat pathway transport is determined by the sequence of the signal peptide, the composition of the Tat apparatus, and the intrinsic characteristics of exported proteins. PMID- 11922669 TI - Bacteriophage Mu genome sequence: analysis and comparison with Mu-like prophages in Haemophilus, Neisseria and Deinococcus. AB - We report the complete 36,717 bp genome sequence of bacteriophage Mu and provide an analysis of the sequence, both with regard to the new genes and other genetic features revealed by the sequence itself and by a comparison to eight complete or nearly complete Mu-like prophage genomes found in the genomes of a diverse group of bacteria. The comparative studies confirm that members of the Mu-related family of phage genomes are genetically mosaic with respect to each other, as seen in other groups of phages such as the phage lambda-related group of phages of enteric hosts and the phage L5-related group of mycobacteriophages. Mu also possesses segments of similarity, typically gene-sized, to genomes of otherwise non-Mu-like phages. The comparisons show that some well-known features of the Mu genome, including the invertible segment encoding tail fiber sequences, are not present in most members of the Mu genome sequence family examined here, suggesting that their presence may be relatively volatile over evolutionary time. The head and tail-encoding structural genes of Mu have only very weak similarity to the corresponding genes of other well-studied phage types. However, these weak similarities, and in some cases biochemical data, can be used to establish tentative functional assignments for 12 of the head and tail genes. These assignments are strongly supported by the fact that the order of gene functions assigned in this way conforms to the strongly conserved order of head and tail genes established in a wide variety of other phages. We show that the Mu head assembly scaffolding protein is encoded by a gene nested in-frame within the C terminal half of another gene that encodes the putative head maturation protease. This is reminiscent of the arrangement established for phage lambda. PMID- 11922670 TI - Comparative thermodynamics for monomer and dimer sequence-dependent binding of a heterocyclic dication in the DNA minor groove. AB - Phenylamidine cationic groups linked by a furan ring (furamidine) and related symmetric diamidine compounds bind as monomers in the minor groove of AT sequences of DNA. DB293, an unsymmetric derivative with one of the phenyl rings of furamidine replaced with a benzimidazole, can bind to AT sequences as a monomer but binds more strongly to GC-containing minor-groove DNA sites as a stacked dimer. The dimer-binding mode has high affinity, is highly cooperative and sequence selective. In order to develop a better understanding of the correlation between structural and thermodynamic aspects of DNA molecular recognition, DB293 was used as a model to compare the binding of minor-groove agents with AT and mixed sequence DNA sites. Isothermal titration calorimetry and surface plasmon resonance results clearly show that the binding of DB293 and other related compounds into the minor groove of AT sequences is largely entropy driven while the binding of DB293 as a dimer into the minor groove of GC containing sequences is largely enthalpy-driven. At 25 degrees C, for example, the AT binding has DeltaG degrees, DeltaH degrees and TDeltaS degrees values of 9.6, -3.6 and 6.0 kcal/mol while the values for dimer binding to a GC-containing site are -9.0, -10.9 and -1.9 kcal/mol (per mol of bound compound), respectively. These results show that the thermodynamic components for binding of compounds of this type to DNA are very dependent on the structure, solvation and sequence of the DNA binding site. PMID- 11922671 TI - Multi-resolution contour-based fitting of macromolecular structures. AB - A novel contour-based matching criterion is presented for the quantitative docking of high-resolution structures of components into low-resolution maps of macromolecular complexes. The proposed Laplacian filter is combined with a six dimensional search using fast Fourier transforms to rapidly scan the rigid-body degrees of freedom of a probe molecule relative to a fixed target density map. A comparison of the docking performance with the standard cross-correlation criterion demonstrates that contour matching with the Laplacian filter significantly extends the viable resolution range of correlation-based fitting to resolutions as low as 30 A. The gain in docking precision at medium to low resolution (15-30 A) is critical for image reconstructions from electron microscopy (EM). The new algorithm enables for the first time the reliable docking of smaller molecular components into EM densities of large biomolecular assemblies at such low resolutions. As an example of the practical effectiveness of contour-based fitting, a new pseudo-atomic model of a microtubule was constructed from a 20 A resolution EM map and from atomic structures of alpha and beta tubulin subunits. PMID- 11922672 TI - HIV-1 nucleocapsid protein activates transient melting of least stable parts of the secondary structure of TAR and its complementary sequence. AB - The nucleocapsid protein NCp7 of HIV-1 possesses a nucleic acid chaperone activity that is critical in minus and plus strand transfer during reverse transcription. The minus strand transfer notably relies on the ability of NCp7 to destabilize the stable stem with five contiguous, double-stranded segments of both the TAR sequence at the 3' end of the viral genome and the complementary sequence, cTAR, in minus strong-stop DNA. In order to examine the nature and the extent of NCp7 destabilizing activity, we investigated, by absorbance and fluorescence spectroscopy, the interaction of TAR and cTAR with a (12-55)NCp7 peptide containing the zinc-finger motifs but lacking the ability to aggregate the oligonucleotides. The absorbance changes in the UV band of cTAR show that seven to eight base-pairs, on average, are melted per oligonucleotide at a ratio of one peptide to 7.5 nucleotides. In contrast, the melting of TAR does not exceed an average of one base-pair per oligonucleotide. This may be linked to the greater stability of TAR, since a strong correlation between NCp7 destabilizing effect and oligonucleotide stability was observed. The effect of (12-55)NCp7 on the stem terminus was investigated by using a cTAR molecule doubly labeled at the 3' and 5' ends by a donor/acceptor couple. In the absence of the peptide, about 80 % of the oligonucleotides are in a dark non-fluorescent state, having a close proximity of the two dyes. The remaining 20 % are distributed between three fluorescent species, having either the terminal segment, the two terminal segments or all segments of the stem melted. This is in line with a fraying mechanism wherein the stem terminus fluctuates rapidly between open and closed states. Addition of (12-55)NCp7 shifts the equilibrium toward the open species, suggesting that NC enhances fraying of the stem terminus. Taken together, our data suggest that NCp7 activates the transient opening of base-pairs in the least stable parts of the stem. Also, this activity of NCp7 was found to be dependent on the zinc-finger motifs, since no melting was observed with a fingerless NCp7 peptide. PMID- 11922673 TI - 20 S proteasomes are imported as precursor complexes into the nucleus of yeast. AB - The mechanism by which yeast 20 S proteasomes are imported into the nucleus is still unresolved. Here, we provide the first evidence that 20 S proteasomes are imported as precursor complexes into the nucleus. By using the srp1-49 mutant which is deficient in nuclear import of cargos with classical nuclear localization sequences (cNLS), we show that proteasome precursor complexes associate with importin/karyopherin alphabeta, the cNLS receptor, and that they accumulate inside the cytoplasm. Reconstitution assays revealed that only precursor complexes are targeted to the nuclear envelope (NE) by karyopherin alphabeta. In support, the green fluorescent protein (GFP)-labelled maturation factor Ump1, marking precursor complexes, mainly localizes to the nucleus and around the NE. Our data suggest that nuclear 20 S proteasomes are finally matured inside the nucleus. PMID- 11922674 TI - Solution structure of the N-terminal domain of a potential copper-translocating P type ATPase from Bacillus subtilis in the apo and Cu(I) loaded states. AB - A putative partner of the already characterized CopZ from Bacillus subtilis was found, both proteins being encoded by genes located in the same operon. This new protein is highly homologous to eukaryotic and prokaryotic P-type ATPases such as CopA, Ccc2 and Menkes proteins. The N-terminal region of this protein contains two soluble domains constituted by amino acid residues 1 to 72 and 73 to 147, respectively, which were expressed both separately and together. In both cases only the 73-147 domain is folded and is stable both in the copper(I)-free and in the copper(I)-bound forms. The folded and unfolded state is monitored through the chemical shift dispersion of 15N-HSQC spectra. In the absence of any structural characterization of CopA-type proteins, we determined the structure of the 73-147 domain in the 1-151 construct in the apo state through 1H, 15N and 13C NMR spectroscopies. The structure of the Cu(I)-loaded 73-147 domain has been also determined in the construct 73-151. About 1300 meaningful NOEs and 90 dihedral angles were used to obtain structures at high resolution both for the Cu(I)-bound and the Cu(I)-free states (backbone RMSD to the mean 0.35(+/-0.06) A and 0.39(+/ 0.07) A, respectively). The structural assessment shows that the structures are accurate. The protein has the typical betaalpha(betabeta)alphabeta folding with a cysteine in the C-terminal part of helix alpha1 and the other cysteine in loop 1. The structures are similar to other proteins involved in copper homeostasis. Particularly, between BsCopA and BsCopZ, only the charges located around loop 1 are reversed for BsCopA and BsCopZ, thus suggesting that the two proteins could interact one with the other. The variability in conformation displayed by the N terminal cysteine of the CXXC motif in a number of structures of copper transporting proteins suggests that this may be the cysteine which binds first to the copper(I) carried by the partner protein. PMID- 11922675 TI - Interaction of the myogenic determination factor myogenin with E12 and a DNA target: mechanism and kinetics. AB - The myogenic determination factors MyoD, myogenin, myf5, and MRF4 are members of the basic helix-loop-helix (bHLH) family of transcription factors and crucial agents of myogenesis. The bHLH regions of these proteins enable them to dimerize with E proteins, another class of the bHLH family, and to bind a specific DNA element known as an E box (CANNTG consensus sequence), which results in the activation of muscle-specific gene expression. As a model for such assembly of the myogenic determination factor/E protein-DNA ternary complex, we have studied the physiologically relevant association of myogenin, E12, and the 3' E box of the acetylcholine receptor (AChR) alpha-subunit gene enhancer. Using the technique of electrophoretic mobility shift assay combined with order-of-addition and time-course experiments, we find that heterodimerization of myogenin with E12 occurs prior to DNA-binding. In addition, we deduce the dissociation (Kd) and rate (k) constants for each step in the formation of the myogenin/E12-DNA ternary complex. Kinetic simulations indicate that at 37 degrees C myogenin and E12 heterodimerize with a Kd of 36 microM (k(on) of 573 M(-1) x s(-1) and k(off )of 0.0205 x s(-1)), and that subsequently the heterodimer binds the AChR alpha subunit gene enhancer 3' E box with a Kd of 8.8 nM (with possible k(on) and k(off) values ranging from 1.0x10(8) to 14.1x10(8) M(-1) x s(-1), and 0.875 to 12.3 s(-1), respectively). PMID- 11922676 TI - The crystal structure of the sorcin calcium binding domain provides a model of Ca2+-dependent processes in the full-length protein. AB - Sorcin is a 21.6 kDa calcium binding protein, expressed in a number of mammalian tissues that belongs to the small, recently identified penta-EF-hand (PEF) family. Like all members of this family, sorcin undergoes a Ca2+-dependent translocation from cytosol to membranes where it binds to target proteins. For sorcin, the targets differ in different tissues, indicating that it takes part in a number of Ca2+-regulated processes. The sorcin monomer is organized in two domains like in all PEF proteins: a flexible, hydrophobic, glycine-rich N terminal region and a calcium binding C-terminal domain. In vitro, the PEF proteins are dimeric in their Ca2+-free form, but have a marked tendency to precipitate when bound to calcium. Stabilization of the dimeric structure is achieved by pairing of the uneven EF-hand, EF5. Sorcin can also form tetramers at acid pH. The sorcin calcium binding domain (SCBD, residues 33-198) expressed in Escherichia coli was crystallized in the Ca2+-free form. The structure was solved by molecular replacement and was refined to 2.2 A with a crystallographic R factor of 22.4 %. Interestingly, the asymmetric unit contains two dimers. The structure of the SCBD leads to a model that explains the solution properties and describes the Ca2+-induced conformational changes. Phosphorylation studies show that the N-terminal domain hinders phosphorylation of SCBD, i.e. the rate of phosphorylation increased twofold in the absence of the N-terminal region. In addition, previous fluorescence studies indicated that hydrophobic residues are exposed to solvent upon Ca2+ binding to full-length sorcin. The model accounts for these data by proposing that Ca2+ binding weakens the interactions between the two domains and leads to their reorientation, which exposes hydrophobic regions facilitating the Ca2+-dependent binding to target proteins at or near membranes. PMID- 11922677 TI - Nucleation and propagation of the collagen triple helix in single-chain and trimerized peptides: transition from third to first order kinetics. AB - The kinetics of triple helix formation from single non-crosslinked peptide chains were studied for the collagen models (ProProGly)10 and (ProHypGly)10 in a broad concentration range and compared with those in nucleated trimers. At very low peptide concentrations the reaction order is 3 but decreases at higher concentrations. For (ProProGly)10 the third order rate constant is 800 M(-2) x s( 1) at 7 degrees C, which corresponds to a very long half time of 15 hours at 60 microM chain concentration. For (ProHypGly)10 the rate constant is about 1000 fold higher, which is consistent with the stabilizing effect of 4-hydroxyproline in collagens. The concentration dependence of the reaction order is explained by a nucleation mechanism in which a very unstable dimer is in fast equilibrium with the monomeric chains and addition of the third chain occurs in a rate-limiting step. At high concentrations nucleation is faster than propagation of helix formation and propagation becomes rate-limiting. To test this hypothesis an artificial nucleus was introduced by fusion of (ProProGly)10 with the trimeric foldon domain of T4 phage or the crosslinking domain of collagen III GlyProProGlyProCysCysGlyGlyGly. These domains were recombinantly attached to the C terminus of (GlyProPro)10 and link the three chains in a similar way to the C terminal propeptide domain in collagen III. This results in a local intrinsic chain concentration of about 1 M. A first order reaction is observed for the folding of the triple helix in (GlyProPro)10foldon with a half time of 8.3 minutes, which approximately matches the rate of folding from single chains at 1 M peptide concentration. A high activation energy of 54 kJ/mol is found for this reaction, whereas the temperature dependence of the nucleation step is close to zero, confirming earlier findings on natural collagens that cis-trans isomerization of peptide bonds is the rate-limiting step in propagation. PMID- 11922678 TI - Conformational change of the E-F interhelical loop in the M photointermediate of bacteriorhodopsin. AB - The conformation of the structured EF interhelical loop of bacteriorhodopsin and its change in the M photointermediate were assessed by measuring the rate of reaction of 16 single engineered cysteine residues along the loop with water soluble sulfhydryl reagents. The exposure to the bulk in the unilluminated state determined with the cysteine reaction correlated well with the degree of access to water calculated from the crystallographic structure of the loop. The EF-loop should be affected by the well-known outward tilt of helix F in the M and N intermediates of the photocycle. A second mutation in each cysteine mutant, the D96N residue replacement, allowed full conversion to the M state by illumination. The reaction rates measured under these conditions indicated that buried residues tend to become more exposed, and exposed residues become more buried in M. This is to be expected from tilt of helix F. However, the observation of increased exposure of four residues near the middle of the loop, where steric effects are only from other loop residues, indicate that the conformation of the EF-loop itself is changed. Thus, the motion of the loop in M is more complex than expected from simple tilt of helix F, and may include rotation that unwinds its twist. PMID- 11922694 TI - Peer review of manuscripts. AB - Analysis of the reproducibility of peer review of manuscripts by means of the kappa statistic is fatally flawed from the point of view of statistical theory. An alternative, simple, method of analysis is proposed. On this basis, agreement among reviewers for the Journal of Clinical Neuroscience is at least as good as that reported recently for other clinical neuroscience journals. Nevertheless, a broad review of peer review processes demonstrates that they are far from satisfactory. Might electronic publishing of scientific articles provide a solution? PMID- 11922695 TI - Moving targets and ghosts of the past: outcome measurement in brain tumour therapy. AB - Evaluation of novel therapies for brain tumours should logically consider quality and quantity of patient survival as primary endpoints. The urgency of the problem, however, frequently leads investigators to use surrogate endpoints and historical controls in order to more rapidly evaluate outcome. To examine the impact of the use of surrogate endpoints and historical controls on the evaluation of innovative brain tumour therapy, selective literature review of three content areas (intraarterial chemotherapy for malignant glioma, interstitial brachytherapy for malignant glioma and stereotactic radiosurgery for cerebral metastasis and malignant glioma) was carried out. The impact of surrogate outcome measures and use of historical controls was assessed by comparing the results of trials using these methods and randomised clinical trials. In the evaluation of both intraarterial chemotherapy and interstitial brachytherapy, promising results in early phase trials were not confirmed in randomised clinical trials. This result can be explained by selection bias and predicted by the use of controls carefully selected from large treatment data bases. In the evaluation of stereotactic radiosurgery, early phase trials are promising, but randomised clinical trials have not yet been done. Prior experience suggests that the early promising results with stereotactic radiosurgery should be subjected to randomised clinical trial validation before being considered proven. Careful selection of controls for early phase trials is necessary if erroneous conclusions are to be avoided. PMID- 11922696 TI - Ischaemic brain oedema. AB - Ischaemic brain oedema appears to involve two distinct processes, the relative contribution and time course of which depend on the duration and severity of ischaemia, and the presence of reperfusion. The first process involves an increase in tissue Na+ and water content accompanying increased pinocytosis and Na+, K+ ATPase activity across the endothelium. This is apparent during the early phase of infarction and before any structural damage is evident. This phenomenon is augmented by reperfusion. A second process results from a more indiscriminate and delayed BBB breakdown that is associated with infarction of both the parenchyma and the vasculature itself. Although, tissue Na+ level still seems to be the major osmotic force for oedema formation at this second stage, the extravasation of serum proteases is an additional potentially deleterious factor. The relative importance of protease action is not yet clear, however, degradation of the extracellular matrix conceivably leads to further BBB disruption and softening of the tissue, setting the stage for the most pronounced forms of brain swelling. A number of factors mediate or modulate ischaemic oedema formation, however, most current information comes from experimental models, and clinical data on this microcosmic level is lacking. Clinically significant brain oedema develops in a delayed fashion after large hemispheric strokes and is a cause of substantial mortality. Neurological signs appear to be at least as good as direct ICP measurement and neuroimaging in detecting and gauging the secondary damage produced by stroke oedema. The neuroimaging characteristics of the stroke, specifically the early involvement of greater than half of the MCA territory, are, however, highly predictive of the development of severe oedema over the subsequent hours and days. None of the available medical therapies provide substantial relief from the oedema and raised ICP, or at best, they are temporizing in most cases. Hemicraniectomy appears most promising as a method of avoiding death from brain compression, but the optimum timing and manner of patient selection are currently being investigated. All approaches to massive ischaemic brain swelling are clouded by the potential for survival with poor functional outcome. It is possible to manage blood pressure, serum osmolarity by way of selective fluid administration, and a number of other systemic factors that exaggerate brain oedema. Broad guidelines for treatment of stroke oedema can therefore be given at this time. PMID- 11922697 TI - Mentors in neuroscience. Peter Orlebar Bishop. PMID- 11922698 TI - Relation of lipid and lipoprotein(a) to ischaemic stroke. AB - The relationship of lipids and Lp(a) to ischemic stroke hasn't been established yet. Our aim was to determine lipid profile and vascular risk factors in stroke patients and compare them with control subjects. Seventy-nine consecutive patients with ischemic stroke were analyzed by total cholesterol, HDL-C, LDL-C, triglyceride, Lp(a) and doppler ultrasonography and vascular risk factors were recorded. Thirty control subjects of same ages were compared with the patient group. Lp(a) and lipids were correlated with stroke subtype and carotid atherosclerosis. There was no statistical significance between patients and control subjects related to total cholesterol, triglyceride, HDL-C, LDL-C and Lp(a) (P>0.05). Atherotrombotic and lacunar strokes didn't show any difference correlated with lipids and Lp(a). Hypertension and diabetes mellitus were important risk factors with (OR=4.50, 95% CI=1.25-16.22) and (OR=4.43, 95% CI=1.79-10.93) respectively. These results were statistically significant (P<0.05). Total cholesterol (308.67+/-85.82) and Lp(a) (32.10+/-17.30) values showed statistical significance (P<0.05) in patients with marked stenosis when compared with patients of normal doppler ultrasonography. Hypertension and diabetes mellitus were found as independent risk factors for ischemic stroke. Lipids and Lp(a) were not independent for atherotrombotic and lacunar stroke. Lp(a) concentration and carotid atherosclerosis in ultrasonography were associated significantly. PMID- 11922699 TI - Cytokine secretion by peripheral blood mononuclear cells in myasthenia gravis. AB - We studied spontaneous secretion of anti-acetylcholine receptor antibody (AChRAb), IgG and cytokines by peripheral blood mononuclear cells (PBMC) from 19 MG patients without therapy and 10 normal controls. IgG secretion was higher in the culture medium of MG than in that of normal controls. AChRAb secretion was correlated with IgG secretion in MG. Interleukin (IL)-2, IL-4, IL-5, IL-6, IL-10, IL-13, interferon (IFN)-gamma and tumor necrosis factor (TNF)-alpha secreted by PBMC from MG patients were not different from those produced by those from normal controls. IgG secretion was, however, correlated with the secretions of IL-5 and IL-6 in MG. Spontaneous B cell activation was suspected in patients with MG. PMID- 11922700 TI - Tumours around the foramen of Monro: clinical and neuroimaging features and their differential diagnosis. AB - The clinical and neuroimaging features of 20 patients with lateral ventricular tumours located around the foramen of Monro were reviewed retrospectively with special emphasis on the differential diagnoses. Histologic types were: eight neurocytomas, four subependymal giant cell astrocytomas (SGCAs), three subependymomas, two fibrillary astrocytomas, and one each of pilocytic astrocytoma, malignant astrocytoma and malignant teratoma. The mean age of the patients with neurocytoma was 29.6 years, with SGCA 13.3 years and with subependymoma 55.3 years. All tumours appeared nodular in shape, and on computed tomography (CT) neurocytomas were either isodense or highdense with the brain, while all subependymomas and SGCAs were lowdense. Calcification was observed in two SGCAs, and one neurocytoma. Five neurocytomas and all four SGCAs showed mild to moderate contrast enhancement, while all three subependymomas showed either no, or scarce, enhancement. Magnetic resonance imaging (MRI) studies were available in 10 patients, with the signal characteristics of four neurocytomas and three SGCAs being nonspecific, while two subependymomas were both hypointense on T1-weighted images and hyperintense on T2-weighted images. Thus important features for differential diagnosis included age of the patient and density on precontrast CT. In this series, either an extensive excision of the tumour or a partial removal, thus relieving the obstruction of the foramina of Monro, usually provided long term survival, with 18 patients surviving a mean of 10.8 years. PMID- 11922701 TI - Resective surgery in infants and young children with intractable epilepsy. AB - Despite the relatively high incidence of epilepsy during the first few years of life, and its documented adverse impact on development, few studies have documented the long term outcomes following epilepsy surgery in infants and young children. We report a consecutive series of 11 patients under 3 years of age who underwent excisional surgery for intractable seizures. A total of 16 resective procedures were performed, and comprised functional hemispherectomy, temporal lobectomy, and lesionectomy. The mean follow-up period was 3 years. At follow-up, 73% (8/11) were seizure-free, with over half of these patients able to cease anti epileptic medications. Acceleration of neurodevelopment was seen in all patients in whom preoperative developmental delay was present. Quality of life was enhanced in all patients. This series confirms that excellent results can be obtained following excisional surgery in well-selected infants and young children with intractable seizures. PMID- 11922702 TI - Clinical characteristics of dural arteriovenous fistula. AB - Intracranial dural arteriovenous fistula (DAVF) is an uncommon neurosurgical condition; in particular, it has been infrequently reported in Korea. To understand the general clinical characteristics of DAVFs, the authors reviewed 53 cases and analyzed factors affecting DAVF hemorrhage of and treatment outcome. Since 1980 we have encountered 480 pial and 53 DAVFs, a ratio of 9.1 to 1. The age of these patients ranged from 1 month to 71 years, the most common being in the 6th decade, and females exceeded males by 1.65 to 1. All lesions except three were single, and symptoms were related to location and the venous drainage pattern. The most common location was the cavernous sinus, accounting for about 64% of cases, with the result that the most common clinical symptoms of DAVFs were ocular, namely proptosis and chemosis. The next was tinnitus also found in transverse-sigmoid sinus DAVFs. Intracranial hemorrhage was seen in eight cases,(15%) the primary cause of hemorrhage was retrograde intracranial venous drainage (P=0.017), and one hemorrhage was observed in cases with no intracranial venous drainage. Intracranial hemorrhage was more frequently in transverse sigmoid than cavernous sinus DAVFs (P=0.049), and this proved to be so even where there was intracranial venous drainage. However, two of 34 patients with cavernous DAVFs became blind in one eye, demonstrating that in such patients, the clinical course could be aggressive. Thirteen patients were treated conservatively. The conservative treatment group was comprised of 13 patients, two of three patients with transverse-sigmoid sinus DAVF expired, and 7 of 10 with cavernous sinus DAVF experienced a clinical improvement or cure. Surgical excision was performed in only two patients. A total of 39 patients underwent embolization; clinical cure was achieved in 13, improvement of symptoms in 12, an unchanged or aggravated result occurred in 9, one died, and four were lost to follow up. During intervention, there was one hemorrhagic complication, owing to obstruction of the venous outflow with embolic materials. In this study, the most common location of DAVFs was the cavernous sinus. The cortical venous drainage remains the primary determinant of intracranial hemorrhage. Common indications for treatment include hemorrhage and neurological deficit. Endovascular treatment is preferred in the majority of cases except tentorial DAVF. The goal of embolization in cavernous DAVF is the alleviation of symptoms, not angiographic cure. But transverse-sigmoid sinus DAVF with venous restriction and leptomeningeal drainage should be treated aggressively. PMID- 11922703 TI - Treatment of high risk or recurrent meningiomas with hydroxyurea. AB - Recurrent, irresectable meningioma constitutes an uncommon but significant problem. Many systemic therapies have been tested without clear evidence of efficacy. More recently, two reports have suggested that hydroxyurea has activity in this context. This study examined the efficacy and toxicity of hydroxyurea in the treatment of 15 patients with high risk meningioma, residual meningioma post resection and progressive meningioma. Hydroxyurea was well tolerated although two patients ceased therapy because of skin rashes. There were no objective responses. Eleven patients achieved stable disease including eight patients who had documented progression prior to commencing hydroxyurea. These results are consistent with previous reports. In conclusion, hydroxyurea may provide some clinical benefit in patients with progressive meningioma by delaying progression of disease. PMID- 11922705 TI - Images in neuroscience: 2. Adrenoleukodystrophy. PMID- 11922704 TI - The neurological abnormalities and operative findings in the transcallosal approach for large juxtasellar-ventricular craniopharyngiomas. AB - From February 1994 to June 1999, there were 36 patients who underwent microsurgical resection for craniopharyngiomas. Of6 adult patients, large tumours (>4 cm) with extension from intrasellar or parasellar space to the foramen of Monro were found. There were 4 male and 2 female patients with ages from 18 to 44 years (mean 32.4 years). All of them underwent one-stage interhemispheric transcallosal total exc ision of tumours. This approach affords good overhead view of tumours and surrounding structures.Preoperatively, decreased visual acuity and restriction in the visual field was found in 5 patients. Studies of anterior pituitary function revealed far below the normal range in all patients. Polyuria was noted in 4 patients. All of 6 patients showed abnormally high levels of blood cholesterol and triglyceride and 4 patients were obese. Tumours with extension into the pituitary fossa were found in 2 cases. A relatively caudal location in the third Ventricle was found in case 6, who showed better preoperative visual function and no polyuria. There were no major surgical complications encountered. One patient with high cholesterol (731 mg%) died of acute pancreatitis 6 months after surgery. For large craniopharyngiomas without pre-chiasmatic extension, proper treatment could be achieved under substitution therapy and one stage transcallosal total excision of tumours with preservation of important neural structures. PMID- 11922706 TI - Microtubule-associated protein 2 (MAP-2) is expressed in low and high grade diffuse astrocytomas. AB - The expression of neuronal antigens in diffuse astrocytomas has not been thoroughly evaluated. We have investigated the expression of microtubule associated protein 2 (MAP-2), synaptophysin and non-phosphorylated epitopes of neurofilament protein (NFP) by immunohistochemistry in 15 low grade diffuse astrocytomas and 15 glioblastomas. MAP-2 was strongly expressed in 97% of cases, using an antibody to both low and high molecular weight isoforms. An antibody specific to high molecular weight isoforms of MAP-2 (hmw-MAP-2) revealed weaker, focal staining in 60% of cases with greater expression in the glioblastomas (P=0.027). NFP was expressed in 50% of cases, but was generally weak and focal. There was little evidence of synaptophysin expression. We conclude that MAP-2 expression in astrocytomas is due predominantly to low molecular weight isoforms, which may be expressed in astrocytes as well as neurons. Focal expression of hmw MAP-2 and NFP, however, suggest that neuronal antigens may be expressed, particularly in high grade astrocytomas. Immunopositivity for these antigens should not preclude the diagnosis of diffuse astrocytoma. PMID- 11922707 TI - Deep brain stimulation of the pedunculopontine region in the normal non-human primate. AB - Lesions in the pedunculopontine nucleus (PPN) cause akinesia. The PPN degenerates in Parkinson's disease. Stimulation of the PPN region induces stepping movements in rats and cats. These findings suggest that the PPN may play an important role in akines ia and that stimulating it may alleviate akinesia.Therefore, we have stereotactically implanted a macroelectrode in the left PPN region in a normal macaque to investigate the effects of deep brain stimulation at different frequencies. Motor activity was recorded with an infra-red counter and behaviour videotaped. At frequencies above 45 Hz there was significantly more severe akinesia than at lower frequencies (paired t -test, n=15, P<0.005). At 100 Hz, there was gross impairment of postural control. At low frequencies (5-30 Hz), stimulation induced a 5-Hz tremor in the right arm. We conclude that stimulating the PPN region at high frequency causes akinesia, whereas low frequencies induce some positive motor effects. PMID- 11922708 TI - MRI evaluation of the diaphragmal opening: using MRI parallel to the transsphenoidal surgical approach. AB - Twenty-six adult diaphragma sellae and infundibulae were examined by MRI parallel to the transsphenonidal surgical plane with attention given to the diaphragmal opening. The diaphragmal opening was observed in 11 cases (42.3%). The anteroposterior diame ter of the opening ranged from 4.0 to 14.0 mm (mean 8.8 mm), and the lateral diameter ranged from 6.0 to 14.0 mm (mean 9.5 mm). In the cases of open diaphragma sellae, the infundibulum tended to be located in the posterior part of the diaphragma sellae but this was not statisticallysignificant. On MRI parallel to the transsphenoidal surgical approach, the anatomy of the dia phragma sellae was well evaluated. PMID- 11922709 TI - Identification of treated target points for parkinsonism on gamma knife follow-up MR images. AB - OBJECT: Identification of treated target points for Parkinsonism on follow-up MR images is difficult because of different orientations employed between the treatment and the follow-up MR scan. In the present work, the treated target points for Parkinsonism can be easily found, once the anterior-commissure (AC) and posterior-commissure (PC) have been defined in Leksell GammaPlan. METHODS: The follow-up MR images must first be defined with the Cartesian co-ordinate system using a non-fiducial based technique. The mathematics of 3-dimensional coordinate geometry is then applied to locate the treated target points for Parkinsonism. Spreadsheet computer software helped to calculate the exact coordinates of the treated target points quickly and accurately. CONCLUSIONS: The coordinates of treated target points can be found easily based on the coordinates of the AC and PC points on the follow-up MR images. Different orientations employed between the treatment and the follow-up MR scan are no longer a problem. PMID- 11922710 TI - Accuracy of sampling methods in morphometric studies of human sural nerves. AB - The aim of this study was to ascertain the minimum sample required to accurately measure the total number of myelinated fibres, mean myelinated fibre density (MFD), myelinated fibre diameter (Ds) and axonal diameter (Da) in morphometric studies of sural nerve biopsies. Measurements were obtained by sampling a single fascicle or systematic sampling of up to 50% of the total transverse fascicular area of two control and eighteen pathological sural nerves showing varying degrees of demyelination and axonal degeneration. MFD and fibre size were heterogeneous between fascicles in both control and pathological sural nerves, and morphometric results from one fascicle and systematic sampling of up to 50% of the total transverse fascicular area did not accurately represent the whole myelinated fibre population in the sural nerve. For accurate morphometric data it is necessary to quantitate all the myelinated fibres in the sural nerve. PMID- 11922711 TI - Hyperintense MCA branch sign on FLAIR-MRI. AB - We report three patients with cardioembolic stroke or transient ischaemic attack. Fluid-attenuated inversion recovery (FLAIR) MRI within hours of symptom onset, demonstrated linear hyperintensities on the surface of the cortex corresponding to neurologic deficits. This unusual finding was indicative of MCA branch occlusion that was confirmed or suggested with angiography. Ultra-early evaluation with FLAIR- and diffusion-MRI may help establish the diagnosis of acute ischaemic stroke particularly due to embolic MCA branch occlusion. PMID- 11922712 TI - Non-herpetic fulminant meningoencephalitis with periodic lateralised epileptiform discharges. AB - This report concerns two patients with fulminant meningoencephalitis presenting in status epilepticus with periodic lateralised epileptiform discharges (PLEDs) observed on electroencephalogram. The titer of herpes simplex virus type 1 antibody was not elevated in either serum or cerebrospinal fluid, and acyclovir was not effective for either patient. Corticosteroid therapy was dramatically effective, however, suggesting that autoimmune inflammatory diseases were the underlying systemic disorders. PLEDs can thus be associated with steroid responsive inflammatory meningoencephalitis. PMID- 11922713 TI - Mesial temporal lobe epilepsy with lateral temporal lobe abnormalities in magnetoencephalography and glucose metabolism. AB - Magnetoencephalography (MEG) and positron emission tomography (PET) revealed abnormal findings in the lateral temporal lobe of a 22 year old female with mesial temporal lobe epilepsy. Electroencephalography identified the epileptogenic focus in the left mesial temporal lobe and standard anterior temporal lobectomy resulted in a good surgical outcome. These discrepancies can be explained by the presence of anatomical and functional pathways between the mesial and lateral temporal structures, or pathophysiological abnormalities in both the mesial and lateral temporal lobes. Careful evaluation is necessary for analysis of MEG and PET findings in patients with temporal lobe epilepsy. PMID- 11922714 TI - Basilar invagination and Chiari malformation associated with cerebellar atrophy: report of two treated cases. AB - We report two patients with an unusual complex of anomalies wherein the basilar invagination and Chiari malformation was associated with marked cerebellar atrophy. Both patients presented with relatively severe lower cranial nerve deficits and showed clinical improvement following a posterior foramen magnum bony decompression. The pathogenesis of the anomalies is discussed and the rationale of treatment is analysed. PMID- 11922715 TI - Growth hormone-secreting pituitary adenoma confined to the sphenoid sinus associated with a normal-sized empty sella. AB - We present a case of growth hormone (GH)-secreting ectopic pituitary adenoma confined to the sphenoid sinus associated with a normal-sized empty sella. It has been well known that acromegaly is sometimes associated with an empty sella. However, such a case usually has a macroadenoma and an empty sella that is large. The authors considered the possible mechanisms of the association between a normal-sized empty sella and an ectopic pituitary adenoma in the sphenoid sinus as the following. Primary empty sella existed originally, and the pituitary adenoma developed later. The adenoma extended into the sphenoid sinus because of the pulsatile intracranial cerebrospinal fluid pressure. PMID- 11922716 TI - Total extrusion of a cranial peritoneal shunt per rectum. AB - This case report demonstrates an unusual complication of a cranial peritoneal shunt. The shunt perforated the bowel and was extruded per rectum. This caused significant anxiety for the individual but did not cause any serious morbidity. PMID- 11922717 TI - Operative findings in cases of trigeminal neuralgia without vascular compression: proposal of a different mechanism. AB - Trigeminal neuralgia is known to be caused by vascular compression at the trigeminal root entry zone (REZ) and microvascular decompression provides good outcome in most of cases. However, in some cases, no vascular compression was observed at the REZ. Over the last 2(1/2) years, the first author operated on 53 cases of trigeminal neuralgia with microvascular decompression and encountered nine cases where no offending vessels were noted at or near the REZ. They were divided into two groups: five cases involving an initial operation and four cases involving a second operation. In the former, arachnoid thickening, angulation or torsion of the root axis were common findings. Dissection of thick arachnoid around the root along the whole length reversed the root to be straight and flaccid. Complete pain relief was noted in four of five cases. In one case of atypical pain, constant facial pain remained. In the latter four cases, where the first operations were done more than 4 years before, thick granulation was noted around REZ without new offending vessels in two cases. In the remaining two cases, where no offending vessels were noted in the first operation, thick adhesion of a distal portion of the root with dura on the pyramidal bone was noted. Meticulous dissection of t he whole length of the root was done and complete pain relief was obtained. Delayed but complete pain relief in these nine cases was noted. Based on operative findings, arachnoid thickening or granulomatous adhesion between the root and surrounding structures can cause an abnormal course of the trigeminal nerve root, which causes root angulation and/or torsion. They can also cause pulsatile movement of the trigeminal nerve root. This tethering effect can promote abnormal root stretching force, especially at REZ, which might promote hyperexitability of the nerve. This speculative mechanism suggests that it is important to make the root free along the entire length, especially at its distal portion in cases with no offending vessels. PMID- 11922718 TI - An unusual posterior mediastinal lipoblastoma with spinal epidural extension presenting as a painful suprascapular swelling: case report and a brief review of the literature. AB - Lipoblastoma is a rare benign pediatric neoplasm of fetal-embryonal fat with little risk of recurrence following total microsurgical excision, but it may progress to local invasion or infiltration if not treated surgically. No adjuvant therapies are usually necessary once the tumor is excised. It is best diagnosed on histopathological studies following excision. An unusual posterior mediastinal lipoblastoma in a 2-year-old Omani girl with spinal epidural extension clinically manifested as a progressive painful suprascapular swelling is reported. It was initially construed to be a benign lipoma, but progressively increasing pain and mild imbalance whilst walking with a tendency to fall on the right side prompted neurosurgical referral and eventual total excision without any added morbidity. Interesting clinical and neuroimaging (CT & MRI) findings are presented and discussed, with a brief review of the literature. PMID- 11922719 TI - Aneurysm of the anterior inferior cerebellar artery (AICA) associated with high flow lesion: report of two cases and review of literature. AB - OBJECTIVE AND IMPORTANCE: Although aneurysms of the anterior inferior cerebellar artery (AICA) are rare lesions, their occurrence in combination with high-flow lesions in the same arterial territory is even more striking. Two cases of an AICA aneurysm in combination with a high -flow lesion are described. CLINICAL PRESENTATION: In one case, a 52-year-old female presented with cerebellar syndrome as the result of a left-sided cerebellar tumor. Angiography revealed a highly vascularized tumor and a broad-based aneurysm at the offspring of the left AICA. In the second case, a 17-year-old female presented with a right-sided cerebellar hemorrhage. Angiography revealed a large peripheral AICA aneurysm and a distal arteriovenous malformation (AVM) fed by the AICA. INTERVENTION: In the first case, a left lateral suboccipital craniotomy was performed and a highly vascularized tumor was removed. The AICA aneurysm could not be adequately clipped and was subsequently wrapped with muscle and reinforced with fibrin glue. Pathological examination of the tumor revealed a hemangioblastoma. Five years after surgery, the patient experienced a subarachnoid hemorrhage. Subsequent vertebral angiography revealed local enlargement of the known AICA aneurysm just at the superior aspect, but the patient refused further treatment. In the second case, the patient sustained a novel cerebellar rebleed while awaiting surgery. A right-sided lateral retromastoid suboccipital craniotomy was performed and the AICA aneurysm could be successfully clipped. More peripherally, the AVM with two draining veins could be totally removed. Postoperative angiography revealed no residual aneurysm or AVM. CONCLUSION: Several aspects of these cases are discussed, such as the rare occurrence of AICA aneurysm and the contribution of high-flow lesions to the genesis of the AICA aneurysms. PMID- 11922720 TI - Malignant astrocytoma of the conus medullaris treated by spinal cordectomy. AB - We present a case of malignant astrocytoma of the conus medullaris in a 48-year old man treated by spinal cordectomy. Preoperative examination revealed a tumor at the T12 to L1 level, and intraparenchymal invasion up to T8. The spinal cord was amputated caudally to the root entry zones of the T9 sensory roots. Additional cordectomies were repeated three times because of tumor infiltration at the cut end. At each procedure, the cord was segmentally transected just caudal to the root entry zones of the p reserving-aid sensory roots to minimize the neural deficit. The final transected level was between T3 and T4, and the cut end did not pathologically reveal any tumor invasion. However, the patient died from tumor recurrence and dissemination. Although the attempt to control the tumor by long segment cordectomy was unsuccessful, spinal cordectomy with wide margin may be a possible treatment for patients with malignant astrocytoma of the conus medullaris presenting with complete deficit below the lesion a nd no dissemination, if in an early stage. PMID- 11922721 TI - Localising the source of pneumocephalus: a diagnostic problem. AB - Temporal bone fractures are a common site of origin for pneumocephalus in the trauma setting, but due to the variegated appearance of the skull base, these are not always evident on conventional scanning techniques. We present a case that illustrates this diagnostic difficulty along with the principles of treating pneumocephalus with Ventriculoperitoneal (VP) shunt in situ. PMID- 11922722 TI - Cellular blue naevus of the scalp. PMID- 11922725 TI - Impact load on the triangular fibrocartilage of the wrist: a cadaver study. AB - OBJECTIVE: The aim of this study was to study the role of the triangular fibrocartilage of the wrist in attenuating and transmitting axial force on the ulnar side of the wrist. DESIGN: Ten biopsies from the triangular fibrocartilage of fresh cadaver wrists were subjected to repetitive axial load during 4 h under reproducible conditions. Another five biopsies were subjected to a higher load and for a longer compression time. Finally, five biopsies were compressed at a (three times) higher compression rate. The amount of force transmitted and attenuated as well as the loaded deformation was measured. RESULTS: From the first experiment we concluded that 53% of the axial force was attenuated. More force was attenuated (61%) if the axial load was increased but still kept within the physiological boundaries. However, increasing the compression rate beyond the physiological boundaries showed that only very little force is attenuated (11.2%). CONCLUSION: The triangular fibrocartilage of the wrist has an important force attenuating function and should not be easily resected. PMID- 11922726 TI - Gallium nitrate accelerates partial thickness wound repair and alters keratinocyte integrin expression to favor a motile phenotype. AB - The nitrate form of the Group III transitional element gallium (GN) increases expression of specific structural components of the provisional wound matrix (i.e., collagen type I, fibronectin) in human dermal fibroblasts. To evaluate the potential of GN as a therapeutic option in management of cutaneous trauma, GN treated partial thickness porcine wounds and experimentally "injured" human keratinocyte (NHK) monolayer cultures were compared with mirror image control (i.e., saline-treated) sites. GN suppressed cell proliferation in both models, as determined by reduced Ki-67 reactivity and significant lengthening of keratinocyte cell cycle transit times, while effectively promoting reepithelialization. The primary effect of GN was apparently to promote cell migration, as neither epidermal thickness nor epidermal differentiation was altered as a result of GN exposure in vivo or in vitro. Significantly enhanced epidermal reepithelialization was associated with alterations in expression of several keratinocyte integrin subunits. GN induced a significant increase in alpha5 expression. alpha5beta1 switching is a characteristic of the motile phenotype in the setting of cutaneous injury. Concomitantly, GN treatment also induced a dramatic (70%) decrease in the expression of the alpha3 subunit; alpha3beta1 binds laminin 5 and is associated with hemidesmosome formation and reestablishment of a nonmotile phenotype. Taken together, the GN-induced changes in integrin expression favor acellular migration. While the molecular mechanism of GN action on resident cells of the skin remains to be defined, these data suggest that GN administration which represses MMP activity in the wound and increases matrix synthesis also accelerates NHK motility and, thereby, may be a useful therapeutic agent for wound repair. PMID- 11922728 TI - Tamoxifen decreases fibroblast function and downregulates TGF(beta2) in dupuytren's affected palmar fascia. AB - BACKGROUND: Dupuytren's contracture is a fibroproliferative disorder that is associated with increased collagen deposition. Isoforms of transforming growth factor beta (TGF(beta)), normally TGF(beta1) and TGF(beta2), are involved in the progressive fibrosis of Dupuytren's disease. It has been suggested that downregulation of TGF(beta) may be useful in the treatment of the condition. Tamoxifen, a synthetic nonsteroidal antiestrogen, is known to modulate the production of TGF(beta). This study examined the role of tamoxifen in decreasing fibroblast function and downregulating TGF(beta2). METHODS: Primary cultures of fibroblasts were obtained from Dupuytren's affected fascia and carpal tunnel affected fascia as a control. Collagen lattices were prepared and populated with the fibroblasts. The fibroblast-populated collagen lattices (FPCL) were then measured for contraction every 24 h for 5 days. Supernatant was obtained from the culture medium following completion of the FPCL portion of the experiment and used for a TGF(beta2) immunoassay. RESULTS: Dupuytren's affected fibroblasts contracted the FPCLs significantly more than carpal tunnel control fibroblasts. Treating the fibroblasts with tamoxifen caused a decreased contraction rate in both Dupuytren's affected fibroblasts and carpal tunnel controls. There was increased TGF(beta2) expression in the Dupuytren's affected fascia group compared to the carpal tunnel control group. Tamoxifen decreased TGF(beta2) expression in Dupuytren's affected fascia group but not in the carpal tunnel control group. CONCLUSION: TGF(beta) appears to be the key cytokine in the fibrogenic nature of Dupuytren's disease. Tamoxifen treatment has been demonstrated to decrease the function of fibroblasts derived from Dupuytren's affected fascia and downregulated TGF(beta2) production in these same fibroblasts. These data suggest a method to manipulate and control Dupuytren's contracture in the clinical setting. PMID- 11922727 TI - Lexipafant inhibits postsurgical adhesion formation. AB - BACKGROUND: PAF and its antagonists have been studied in the pathophysiology of various inflammatory conditions. This study investigates the effects of a platelet activating factor antagonist, lexipafant, on peritoneal adhesion formation and wound healing. MATERIALS AND METHODS: Forty-eight Wistar albino rats (300-350 g) were divided into four equal groups; adhesion-induced lexipafant (AL), adhesion-induced saline (AS), sham-operated lexipafant (SL), and sham operated saline (SS). All rats underwent a midline laparotomy under sterile conditions. The anterior wall of the left uterine horn was scraped to cause hemorrhages in adhesion-induced groups. Following peritoneal injections of either saline or lexipafant, the incisions were closed in layers. On the 14th day, the rats were killed and adhesions were scored from 0 (none) to 4 (dense). Tissue samples from the adhesions and the left horn of uterus were examined biochemically for hydroxyproline content, and serum IL-6 levels were determined. RESULTS: The adhesion formation score was significantly increased in the AS group compared to the SL and AL groups (P < 0.001). The IL-6 levels of the AS group were higher than those of the other groups (P < 0.05). There was no significant difference in hydroxyproline content between groups (P > 0.05). CONCLUSIONS: Lexipafant plays a role in the prevention of adhesion formation without affecting wound healing. PMID- 11922729 TI - Late complication of aortoiliac stent placement- atheroembolization of the lower extremities. AB - BACKGROUND: Atheroembolization following aortoiliac stent placement is uncommon. The purpose of this study was to examine the management and risk factors of lower extremity atheroembolization following aortoiliac stent placement for occlusive disease. MATERIALS AND METHODS: From March 1993 to February 2001, the hospital records of all patients who developed thromboembolic events following aortoiliac stent placement were reviewed. Risk factor analysis was performed by comparing with the control group, which consisted of 493 patients treated with aortoiliac stents during the study period who did not develop atheroembolic complications. Patients with cardiac etiologies or aortic aneurysms as the source of embolization as well as those who developed acute embolization following stent deployment (<30 days) were excluded. RESULTS: Atheroembolization occurred in eight patients (12 iliac artery stents and 1 aortic stent) at intervals ranging from 9 to 43 months (mean 22 months) following aortoiliac stent placement. Arteriography in all patients implicated the stented artery as the source of atheroembolism. Five corrective operations (two aorto-bifemoral bypasses, one ileofemoral bypass, and two aortoiliac endarterectomies) along with two concomitant femoropopliteal thrombectomies were performed successfully in five patients. The remaining three patients were treated with either thrombolysis and/or additional stent placement, which resulted in either iliac occlusion or recurrent embolic symptoms (P < 0.05). All 3 patients subsequently underwent bypass procedures (one ileofemoral and two femorofemoral bypasses). There was no perioperative mortality. During a mean follow-up of 16 months (range 3 to 45 months), two patients required minor amputations, whereas one required major leg amputation. No further episodes of atheroembolism occurred in the involved limbs following surgical bypass procedures. Risk factor analysis failed to identify potential variables that correlated with atheroembolism following aortoiliac stent placement. CONCLUSION: Patients with atheromatous embolization following aortoiliac stent placement should be evaluated aggressively. The treatment of choice is surgical correction or bypass with exclusion of the offending embolic source. Although intra-arterial stent placement in the atheroembolic stented iliac artery is feasible, it may provide a less durable result. PMID- 11922730 TI - Advantages of positron emission tomography over computed tomography in mediastinal staging of non-small cell lung cancer. AB - BACKGROUND: New treatment algorithms in early stage non-small cell lung cancer (NSCLC) involving preoperative chemotherapy require accurate clinical staging of the mediastinum. This study compares the accuracy of 2-[fluorine-18]fluoro-2 deoxy-d-glucose (FDG) positron emission tomography (PET) scanning with that of computed tomography (CT) scanning in the clinical staging of non-small cell lung cancer. MATERIALS AND METHODS: A retrospective review was performed on 52 patients with NSCLC who were evaluated with both CT and PET scans. All patients had their mediastinal lymph nodes sampled by mediastinoscopy or at the time of thoracotomy for pulmonary resection. Each imaging study was evaluated separately and correlated with histopathologic results. RESULTS: For detecting mediastinal metastases the sensitivities of PET and CT scans were 67 and 50%, respectively; specificities were 91 and 65%, respectively; accuracies were 88 and 63%, respectively; positive predictive values were 50 and 16%, respectively; negative predictive values were 95 and 88%, respectively. PET scans were significantly better than CT scans at detecting mediastinal metastases (PET, 4/8; CT, 3/19) (P = 0.01). CONCLUSIONS: PET scanning is superior to CT scanning for clinical staging of the mediastinum in NSCLC. A more confident decision regarding stratification of patients into current treatment algorithms can be made when the decision is based on PET scanning rather than the current "gold standard" of CT scanning. PMID- 11922731 TI - Effect of blood flow occlusion on laser hyperthermia for liver metastases. AB - BACKGROUND: Interstitial laser hyperthermia (ILH) is an in situ ablative technique for the treatment of colorectal liver metastases. A significant factor limiting tumor destruction is hepatic blood flow. Modulation of hepatic blood flow may increase the size of tumor necrosis achieved. Our aim was to investigate the effect of blood flow occlusion on ILH-induced necrosis in both tumor and normal liver tissue. MATERIALS AND METHODS: A model of colorectal liver metastases in male inbred CBA mice was used. ILH was applied to normal liver and tumor tissue using a bare optical quartz fiber from an SYL500 Nd:YAG surgical laser generator, with and without hepatic blood flow occlusion, and the extent of necrosis was studied. Tumor blood flow was assessed by laser Doppler flowmetry and scanning electron microscopy. RESULTS: Hepatic blood flow occlusion resulted in a significant reduction in blood flow in normal liver tissue (37.9% +/- 5.8, P < 0.001) and in the periphery of the tumor (17.5% +/- 7.8, P < 0.001). It did not affect the blood flow in the center of the tumor (13.4% +/- 4.3, P = 0.07). ILH of normal liver tissue, at low power (2 W), with hepatic blood flow occlusion, resulted in a significant increase in the diameter of necrosis. This effect was not seen when higher power (5 W) was used in normal liver. No significant effect was noted within tumor tissue at either power setting. CONCLUSION: The overall effect of hepatic blood flow occlusion in ILH-induced tissue necrosis appears to be negligible in tumor tissue. Its main applicability appears to be at the tumor host interface, where a decrease in blood flow may lead to higher temperatures and therefore to a greater degree of tumor cell destruction. PMID- 11922732 TI - Augmentation of wound healing with translation initiation factor eIF4E mRNA. AB - BACKGROUND: Initiation of translation is the rate-limiting step in protein synthesis; eIF4E increases translational efficiency by facilitating ribosome scanning. eIF4E is present in cells in rate-limiting amounts; chronic overexpression of eIF4E causes cell transformation by upregulating growth-related proteins. Biolistic delivery of epidermal growth factor (EGF) increases wound healing; transiently increasing wound eIF4E levels with biolistic mRNA transmission may further augment wound healing without oncogenesis. PATIENTS AND METHODS: Midline fascial wounds were created in rats and biolistically treated with gold particles carrying mRNA encoding for hEGF with or without eIF4E prior to suture closure; control animals received blank bullets. The animals were sacrificed at 7 or 14 days for determination of peak wound bursting strength on a tensiometer. Results are expressed as means +/- standard deviation; statistics were via analysis of variance. RESULTS: [Table: see text]. CONCLUSIONS: Simultaneous biolistic delivery of EGF mRNA with eIF4E mRNA significantly increases wound breaking strength compared to that in control animals or treatment with EGF mRNA alone without risk of cellular transformation. Further studies of translational activation to augment wound healing are warranted. PMID- 11922734 TI - Morphologic study of small intestinal submucosa as a body wall repair device. AB - BACKGROUND: The extracellular matrix (ECM) derived from porcine small intestinal submucosa (SIS) has been used as a constructive scaffold for tissue repair in both preclinical animal studies and human clinical trials. Quantitative characterization of the host tissue response to this xenogeneic scaffold material has been lacking. MATERIALS AND METHODS: The morphologic response to a multilaminate form of the SIS-ECM was evaluated in a chronic, 2-year study of body wall repair in two separate species: the dog and the rat. Morphologic response to the SIS-ECM was compared to that for three other commonly used bioscaffold materials including Marlex mesh, Dexon, and Perigard. Quantitative measurements were made of tissue consistency, polymorphonuclear cell response, mononuclear cell response, tissue organization, and vascularity at five time points after surgical implantation: 1 week, 1, 3, and 6 months, and 2 years. RESULTS: All bioscaffold materials functioned well as a repair device for large ventral abdominal wall defects created in these two animal models. The SIS-ECM bioscaffold showed a greater number of polymorphonuclear leukocytes at the 1-week time point and a greater degree of graft site tissue organization after 3 months compared to the other three scaffold materials. There was no evidence for local infection or other detrimental local pathology to any of the graft materials at any time point. CONCLUSIONS: Like Marlex, Dexon, and Perigard, the SIS-ECM is an effective bioscaffold for long-term repair of body wall defects. Unlike the other scaffold materials, the resorbable SIS-ECM scaffold was replaced by well organized host tissues including differentiated skeletal muscle. PMID- 11922733 TI - Retinoids inhibit squamous cell carcinoma growth and intercellular communication. AB - BACKGROUND: Retinoids have been shown to inhibit the growth of squamous cell carcinoma and other malignancies. They have also been shown to alter gap junctional intercellular communication (GJIC) and the expression of connexins, the protein subunits of gap junctions. We report in this study that the alteration of GJIC by retinoids may be directly related to inhibitory effects on cell growth. MATERIALS AND METHODS: SCC-13 cells were treated with all-trans retinoic acid (tRA) and 13-cis retinoic acid (cRA) at 10(-7) and 10(-6) M concentrations in culture. No treatment and ethanol vehicle controls were included for each experiment. Serial cell counts of parallel cultures were performed to determine cell growth. The parachute technique was performed in combination with fluorescence activated cell sorting (FACS) analysis to determine GJIC. Northern and Western blot analysis were performed to assess connexin mRNA and protein expression. RESULTS: The growth rate was inhibited for cells treated with tRA (10(-6) M) (P < 0.05) and cRA (10(-6) M) (P = 0.068) vs. vehicle control. GJIC was significantly inhibited with both tRA (10(-7) and 10(-6) M) (P < 0.001) and cRA (10(-7) and 10(-6) M) (P < 0.001) at 24, 48, and 96 h as determined by FACS analysis. To correlate GJIC with cell growth, we studied the effect of glycyrrhetinic acid, a known inhibitor of GJIC. Glycyrrhetinic acid also significantly inhibited cell growth (P < 0.05) vs. control. Connexin 26 and connexin 43 mRNA and protein expression were not significantly altered after retinoid treatment. CONCLUSION: Retinoic acids inhibit both cell growth and GJIC in SCC-13 cells. Retinoids may inhibit cell growth through alteration of GJIC in SCC-13 cells. PMID- 11922735 TI - Expression of PH-20 in normal and neoplastic breast tissue. AB - BACKGROUND: Tumor metastasis involves a sequence of interrelated steps, of which penetration beyond the basement membrane is an essential component. Hyaluronic acid (HA) is a major structural component of the complex proteoglycans found in extracellular matrices and basement membranes. Hyaluronidase (PH-20) degrades HA, resulting in the disruption of basement membrane integrity and possible tumor dissemination. MATERIALS AND METHODS: Total RNA was extracted from samples (n = 51) of normal breast tissue (n = 12), ductal carcinoma in situ (DCIS) (n = 12), infiltrating ductal breast adenocarcinoma (n = 13), and metastatic breast cancer to lymph nodes (n = 14). RT-PCR was used to determine the relative level of PH-20 in each specimen. RESULTS: PH-20 was detected in 41/51 (80.4%) of the specimens evaluated. PH-20 was present in 12/12 (100%) normal breast tissues; 8/12 (66.7%) DCIS; 13/13 (100%) invasive breast cancers; and 8/14 (57.1%) metastases. Of those specimens in which PH-20 was detected, there were increased levels of PH-20 in metastatic breast cancer to lymph nodes compared to DCIS and invasive breast cancer. Stratification of specimen by race revealed that African American women had higher levels of PH-20 with invasive and metastatic beast cancer. CONCLUSIONS: Increased levels of PH-20 are noted in invasive and metastatic breast cancer compared to DCIS. Tumors from African American women with invasive and metastatic breast cancer demonstrated higher levels of PH-20 than Caucasians. Varying levels of PH-20 in mammary tissue may contribute to early invasion and metastasis of breast cancer. PMID- 11922736 TI - Impact of polymer pore size on the interface scar formation in a rat model. AB - BACKGROUND: The surgical therapy of hernias is increasingly based on reinforcement with alloplastic material, in particular surgical meshes. The biological response to these foreign bodies largely depends on the selected material and its structure. In comparison to the physiological scar process following a simple abdominal wall incision, the chronic inflammation at the interface to the polymers lead to specific morphological alterations. MATERIALS AND METHODS: In the present study two meshes with different pore sizes were implanted into rats: a heavy-weight and small-pore-sized mesh (hw-mesh) made of nonabsorbable polypropylene monofilaments and a low-weight large-pore-sized mesh consisting of polypropylene and of absorbable polyglactin multifilaments (lw mesh). A suture repair of a laparotomy served as control. After 7, 14, 21, and 90 days the mesh area was analyzed with regard to tissue and cellular response. RESULTS: Over the whole observation period morphometric analysis indicated an improved integration of the lw-mesh with reduction of both inflammation and fibrosis, whereas the hw-mesh induced an intense chronic inflammation concomitant with an intensified bridging scar reaction. On the cellular level these findings correspond to an elevated cell turnover, characterized by increased rates of apoptotic and proliferating cells. In contrast, the tissue reaction to the lw mesh achieved levels almost similar to those of the physiological scaring process in the control group. CONCLUSION: In conclusion, the present data confirm the development of a chronic inflammatory foreign body reaction at the interface to both hw-meshes and lw-meshes; however, the use of lw-meshes showed superior tissue integration. With regard to the quite similar polymer surface the pore size appears to be of major importance in tissue reaction and for the biocompatibility of mesh structures. PMID- 11922737 TI - Roles of the jejunum and ileum in the first-pass effect as absorptive barriers for orally administered tacrolimus. AB - BACKGROUND: The immunosuppressant tacrolimus shows poor and variable bioavailability following oral administration in clinical use. Recently, the hepatic and intestinal metabolisms, or first-pass effect, of tacrolimus have been suggested to be responsible for its bioavailability. In the present study, we investigated the respective contribution of the jejunum and ileum to the first pass effect of tacrolimus in rats. METHODS: The metabolism of tacrolimus in everted sacs of the duodenum, jejunum, and ileum was examined. Tacrolimus was administered intravenously or intraintestinally to sham-operated, jejunum resected, or ileum-resected rats. Blood samples were collected over a 240-min period, and whole-blood tacrolimus concentrations were measured by semiautomated microparticle enzyme immunoassay. The pharmacokinetic parameters of tacrolimus in each group were estimated. RESULTS: The metabolic activity of tacrolimus appeared to be the highest in the everted sacs of the duodenum. The bioavailability of tacrolimus in the jejunum- or ileum-resected rats was higher than that in sham operated controls. On the other hand, the time to peak concentration in the jejunum-resected rats was about twofold slower than those in ileum-resected and sham-operated rats. CONCLUSIONS: These results suggested that the first-pass effect of tacrolimus in the small intestine shows regional differences and the extraction of tacrolimus in the small intestine consists of the amount of extraction in the jejunum and ileum. In addition, the ileum rather than the jejunum as a graft of segmental small bowel transplantation would be useful to avoid the adverse effects of tacrolimus. PMID- 11922738 TI - Functional capacity of the thyroid autograft: an experimental study. AB - BACKGROUND: The aim of this study was to investigate the functional capacity of thyroid autografts after total thyroidectomy in a rabbit model. MATERIAL AND METHODS: Thirty-eight rabbits underwent total thyroidetomy. One of the two thyroid lobes was cut into 1-mm pieces and was introduced intramuscularly into the right quadriceps muscle (group A, n = 10), the right rectus abdominalis muscle (group B, n = 10), and the right sacrodorsalis muscle (group C, n = 10). Another group of 8 rabbits underwent total thyroidectomy without autologous implantation and served as the control group (group D). The animals were observed for 8 weeks with weekly measurements of thyroid hormones. At the end of the 8th week, a scintigram was performed. The autografts were removed 2 days later. RESULTS: The thyroid hormone levels showed a gradual decrease until the 2nd-5th week after implantation, followed by a gradual increase and establishment of euthyroid levels between the 5th and 8th weeks. Respectively, an increase of thyreotropin hormone was noted with maximal values in the 4th week, followed by a gradual decrease until the end of the 8th week. The scintigram at the end of the 8th week revealed the presence of functional thyroid tissue in all cases. Functional thyroid follicles were found in all animals who survived. In 35.7% of the autografts, we noted the development of fibrous tissue and gigantocytic granulomas in the periphery, which could be interpretated as a "foreign body" reaction. CONCLUSION: Thyroid autografts can completely substitute thyroid function after total thyroidectomy. PMID- 11922739 TI - Single vs. dual vessel porcine extracorporeal liver perfusion. AB - BACKGROUND: The use of porcine extracorporeal liver perfusion (PECLP) to provide temporary hepatic support for patients in fulminant hepatic failure has been limited by the fact that individual perfusions can be sustained for only a few hours. Inadequate liver function and/or hemodynamic instability are the major contributing factors for early interruption of PECLP. Recent reports suggest that the choice of single (portal vein only) vs dual (portal vein and hepatic artery) vessel perfusion may influence the duration of perfusion. We hypothesize that PECLP with single vessel perfusion (SVP) is associated with worse liver function and greater hemodynamic instability than PECLP with dual vessel perfusion (DVP). MATERIALS AND METHODS: To eliminate the potentially confounding influences of liver failure and xenograft rejection, liver isografts procured from White Landrace pig donors were perfused by either SVP or DVP via an extracorporeal circuit established with normal White-Landrace pig recipients. The function of perfused livers was evaluated by measuring production of bile and Factors V and VIII, clearance of ammonia and lactate, and extraction of O(2) at baseline and at 0, 1, 3, 6, 12, and 24 h after initiation of PECLP. The impact of PECLP on recipient hemodynamic status was assessed by monitoring BP, heart rate, urine output, O(2) saturation, etc. Among other parameters evaluated were serum albumin and total protein and hepatic release of IL-1beta and nitric oxide to assess their possible contributions to hemodynamic instability. RESULTS: DVP and SVP livers cleared ammonia and lactate similarly. Both approaches were associated with progressive hypoalbuminemia and hypoproteinemia. DVP livers produced more bile and Factor V and were associated with less recipient hypotension and IL 1beta and NO release than SVP livers. CONCLUSIONS: Livers with DVP function better than livers with SVP. The duration of PECLP can be limited by recipient hypotension, although this complication is less severe with DVP than with SVP. PMID- 11922740 TI - Glycerol suppresses proliferation of rat hepatocytes and human HepG2 cells. AB - BACKGROUND: In fulminant hepatic failure (FHF), the ability of surviving hepatocytes to proliferate is diminished. Therefore, it is important that medical therapy cause no further impairment of liver regeneration. In FHF, intracranial hypertension secondary to brain edema is the most common cause of brain injury and death and glycerol is used in some countries to treat this complication. Glycerol has been long known to suppress the growth of various cell types. We therefore decided to examine the effect of glycerol on hepatocyte proliferation in vitro and in vivo in rats subjected to partial (2/3) hepatectomy. Additionally, we investigated the effect of glycerol on the proliferation of HepG2 cells. MATERIALS AND METHODS: Mitogen-induced primary rat hepatocytes were cultured in a hormonally defined Dulbecco's modified Eagle's medium containing increasing amounts of glycerol (0.5, 1.0, 2.0, 4.0%). HepG2 cells were cultured in minimal essential medium/10% FBS. After 2 days, HepG2 cells were exposed to glycerol (1.0-2.0-4.0%) and harvested after 48 h. Control dishes contained no glycerol. Cell proliferation was measured by the incorporation of [(3)H]thymidine and/or bromodeoxyuridine (BrdU). In vivo, Sprague-Dawley rats were subjected to standard partial 2/3 hepatectomy and assigned to intraportal administration of either 400 microl of glycerol or saline. Rats were killed after 1, 2, 3, 5, and 7 days. Liver weight/body weight ratio and BrdU uptake were measured. RESULTS: In all cultures tested, glycerol suppressed the growth of cells in a dose-dependent manner. In vivo, a single intraportal dose of glycerol slowed the liver regenerative response. CONCLUSIONS: This study demonstrated that glycerol has a potent growth-inhibitory effect on hepatocyte proliferation in vivo and in vitro. Remarkably, glycerol inhibited the proliferation of liver cancer cells as well. The results of this study have important clinical implications. PMID- 11922741 TI - Ischemic preconditioning improves mitochondrial tolerance to experimental calcium overload. AB - BACKGROUND: Ca(2+) overload leads to mitochondrial uncoupling, decreased ATP synthesis, and myocardial dysfunction. Pharmacologically opening of mitochondrial K(ATP) channels decreases mitochondrial Ca(2+) uptake, improving mitochondrial function during Ca(2+) overload. Ischemic preconditioning (IPC), by activating mitochondrial K(ATP) channels, may attenuate mitochondrial Ca(2+) overload and improve mitochondrial function during reperfusion. The purpose of these experiments was to study the effect of IPC (1) on mitochondrial function and (2) on mitochondrial tolerance to experimental Ca(2+) overload. METHODS: Rat hearts (n = 6/group) were subjected to (a) 30 min of equilibration, 25 min of ischemia, and 30 min of reperfusion (Control) or (b) two 5-min episodes of ischemic preconditioning, 25 min of ischemia, and 30 min of reperfusion (IPC). Developed pressure (DP) was measured. Heart mitochondria were isolated at end-Equilibration (end-EQ) and at end-Reperfusion (end-RP). Mitochondrial respiratory function (state 2, oxygen consumption with substrate only; state 3, oxygen consumption stimulated by ADP; state 4, oxygen consumption after cessation of ADP phosphorylation; respiratory control index (RCI, state 3/state 4); rate of oxidative phosphorylation (ADP/Deltat), and ADP:O ratio) was measured with polarography using alpha-ketoglutarate as a substrate in the presence of different Ca(2+) concentrations (0 to 5 x 10(-7) M) to simulate Ca(2+) overload. RESULTS: IPC improved DP at end-RP. IPC did not improve preischemic mitochondrial respiratory function or preischemic mitochondrial response to Ca(2+) loading. IPC improved state 3, ADP/Deltat, and RCI during RP. Low Ca(2+) levels (0.5 and 1 x 10(-7) M) stimulated mitochondrial function in both groups predominantly in IPC. The Control group showed evidence of mitochondrial uncoupling at lower Ca(2+) concentrations (1 x 10(-7) M). IPC preserved state 3 at high Ca(2+) concentrations. CONCLUSIONS: The cardioprotective effect of IPC results, in part, from preserving mitochondrial function during reperfusion and increasing mitochondrial tolerance to Ca(2+) loading at end-RP. Activation of mitochondrial K(ATP) channels by IPC and their improvement in Ca(2+) homeostasis during RP may be the mechanism underlying this protection. PMID- 11922742 TI - An evaluation of the effectiveness of extended lymph node dissection in patients with gastric cancer: a retrospective study of 1403 cases at a single institution. AB - BACKGROUND: Many investigators have reported that extended lymph node dissection (D2 dissection) is probably an effective procedure. However, the theory that D2 dissection leads to an improvement in survival has not been confirmed in randomized trials. We attempted to confirm the effectiveness of D2 dissection with gastrectomy for gastric cancer. MATERIALS AND METHODS: Gastric cancer patients (1403) underwent curative resection by D1 (991 patients) or D2 (412 patients) dissection with gastrectomy. Survival rates calculated for all patients and subdivided for stage, depth of invasion, and lymph node metastasis were compared between the two groups. The diagnosis of lymph node metastasis was compared between macroscopic and histological findings. RESULTS: There was no significant difference in the survival of patients overall. However, in the patients with stage II, T1 or T2, or N1 disease, the survival of the D2 group was significantly better than that of the D1 group. The false positive rates of lymph node metastasis were 53.3% in the N1 group, 26.2% in the N2 group, and 9.2% in the N3 group. In a considerable proportion of the N1 and N2 patients, histological findings proved more or fewer metastases than macroscopic diagnosis. CONCLUSIONS: Metastatic lymph nodes should be resected as far as possible. D2 dissection with gastrectomy is recommended for T1, N1 or T2, N1 disease, particularly in younger patients. PMID- 11922743 TI - Protective effect of a new C5a receptor antagonist against ischemia-reperfusion injury in the rat small intestine. AB - BACKGROUND: The complement system is a major contributor to the pathogenesis of intestinal ischemia-reperfusion (I/R) injury. We have studied the action of an orally active complement factor 5a (C5a) receptor antagonist, the cyclic peptide AcF-(OPdChaWR) [Ac-Phe(Orn-Pro-d-cyclohexylalanine-Trp-Arg)] against local and remote intestinal I/R injuries in rats. MATERIALS AND METHODS: Anesthetized rats were administered with AcF-(OPdChaWR) at doses of 1 mg/kg intravenously or 0.3, 1, or 10 mg/kg orally with pyrogen-free saline for sham control animals. The superior mesenteric artery was occluded for 30 min and the intestine reperfused for 120 min. Changes associated with tissue injury were assessed by neutropenia, intestinal edema, serum tumor necrosis factor-alpha, serum haptoglobin, plasma aspartate aminotransferase, and histopathology. RESULTS: Pretreatment with either a single intravenous dose (1 mg/kg), or a single oral dose (10 mg/kg) of AcF (OPdChaWR) significantly inhibited I/R induced neutropenia, the elevated serum levels of tumor necrosis factor-alpha, haptoglobin, and plasma aspartate aminotransferase, as well as intestinal edema. Histological analysis of AcF (OPdChaWR)-treated I/R animals showed markedly reduced mucosal layer damage compared to that of untreated rats. CONCLUSIONS: These results indicate that a potent antagonist of C5a receptors on human cells protects the rat small intestine from I/R injury after oral or intravenous administration. Small molecule C5a antagonists may have some therapeutic utility in reperfusion injury. PMID- 11922744 TI - Periarterial papaverine application improves intraoperative kidney function during laparoscopic donor nephrectomy. AB - BACKGROUND: Laparoscopic donor nephrectomy decreases disincentives to donation frequently associated with the disadvantages of open surgery. However, concerns have been raised regarding graft quality, since the incidence of delayed graft function is higher when compared with open procedures. This may be caused by amelioration of kidney perfusion due to the elevated intraabdominal pressure and to a mechanically induced renal angiospasm during donation. This study was addressed to reveal whether the renal periarterial application of papaverine is able to enhance renal blood flow during laparoscopic nephrectomy. MATERIALS AND METHODS: Twelve male piglets underwent left laparoscopic donor nephrectomy after endoscopic occlusion of the right renal vessels and ureter. Urine output and creatinine clearance were determined as indicators of renal blood flow. In the treatment group (n = 6) papaverine hydrochloride was administered to the tissue surrounding the renal artery prior to preparation of the vessels and results were compared with those of controls (n = 6). Free sodium excretion was measured to preclude prerenal failure. RESULTS: In the control group the mean urine output was 0.015 ml/min/kg and the mean creatinine clearance was 0.95 ml/min/kg. In pigs treated with papaverine the mean urine output was 0.052 ml/min/kg and the mean creatinine clearance was 2.22 ml/min/kg. The differences were significant (urine output, P = 0.02; creatinine clearance, P = 0.038). CONCLUSIONS: Papaverine improves renal function during laparoscopic kidney harvest when applied in the vicinity of the renal artery prior to vascular preparation. PMID- 11922745 TI - Effect of prophylactic intracerebroventricular injection of methylprednisolone on nitrogen and catecholamine excretion in the urine after laparotomy in rats. AB - BACKGROUND: This study was carried out to determine whether the prophylactic injection of glucocorticoid into the intracerebroventricular (i.c.v.) space reduced TNF-alpha and IL-1beta mRNA synthesis in the brain after laparotomy, resulting in a reduction of nitrogen excretion in the urine. PATIENTS AND METHODS: Male SD rats (body wt., 225-250 g, n = 114) were catheterized into the i.c.v. space on day 0. On day 4, the rats were assigned to four groups: (1) Control, (2) laparotomy (Trauma), (3) intraperitoneal (i.p.) injection of methylprednisolone (MP) plus laparotomy (IPMP), and (4) i.c.v. injection of MP plus laparotomy (ICVMP). Either 3 or 24 h after surgery, the animals were sacrificed. TNF-alpha and IL-1beta mRNA levels in tissues, including the brain cortex and hypothalamus, were measured by RT-PCR. The amounts of nitrogen and catecholamine excretion in the 24-h urine were determined. RESULTS: The i.p. injection of MP reduced TNF-alpha and IL-1beta mRNA levels in all the tissues 3 h after laparotomy compared with those of the Trauma group. The icv injection of MP prevented elevation of the TNF-alpha and IL-1beta mRNA levels in the brain (cortex, TNF-alpha, ICVMP 0.43 +/- 0.06, P < 0.05, vs Trauma; cortex, IL-1beta, ICVMP 0.25 +/- 0.09, P < 0.05, vs. Trauma; hypothalamus, TNF-alpha, ICVMP 0.31 +/ 0.04, P < 0.05, vs. Trauma; hypothalamus, IL-1beta, ICVMP 0.25 +/- 0.14, P < 0.05, vs. Trauma), but did not inhibit an increase in TNF-alpha and IL-1beta mRNA levels in the liver and skeletal muscle. Both nitrogen and catecholamine excretions in the urine were decreased by ip and by i.c.v. injection of MP compared to those of the Trauma group (nitrogen, ICVMP 559.3 +/- 52.0 mg/day, P < 0.05, vs. Trauma; catecholamine, ICVMP 13.8 +/- 1.8 microg/day, P < 0.05, vs. Trauma). CONCLUSION: A reduction in TNF-alpha and IL-1beta mRNA synthesis in the brain due to prophylactic injection of MP into the icv space reduced the catabolic response after laparotomy. PMID- 11922746 TI - Angiogenesis induced by the injection of peripheral leukocytes and platelets. AB - BACKGROUND: Peripheral leukocytes and platelets (LAPs) contain many kinds of cells with the ability to secrete several growth factors and cytokines. We attempted to induce therapeutic angiogenesis by injecting self-LAPs into a rat ischemic hindlimb model. MATERIALS AND METHODS: Supernatants from cultured LAPs were used for the endothelial cell (EC) proliferation assay, and LAPs were used in a cornea model to evaluate angiogenic potency. LAPs were injected directly into the male Dark Agouti rat ischemic hindlimb model, after which a microangiogram was done and the capillary/muscle fiber ratio was examined histologically. ELISA revealed the levels of contributing growth factors and cytokines present in the ischemic muscles. RESULTS: The EC proliferation assay showed that the supernatants of LAPs accelerated proliferation and that the LAPs induced angiogenesis in the cornea model. The microangiograms and histological evaluation revealed that angiogenesis was induced more effectively in the rats injected with LAPs (LAP group) than in the those injected with phosphate-buffered saline (PBS group). The levels of basic fibroblast growth factor (bFGF) in the ischemia, PBS, and LAP groups were significantly increased compared to those in the sham group. The level of interleukin-1beta (IL-1beta) in the LAP group was significantly more elevated than in the other groups. CONCLUSIONS: The injection of self-LAPs induced angiogenesis in a rat ischemic hindlimb model. Ischemia caused an elevation in the level of bFGF and also in IL-1beta derived from LAPs, which contributed to angiogenesis. This is a novel, yet simple and safe method of inducing therapeutic angiogenesis. PMID- 11922747 TI - Affinity-purification of Transferrin-binding protein B under nondenaturing conditions. AB - The commonly used purification procedures for Transferrin-binding protein B (TbpB) are based on an affinity chromatography step using resins onto which human transferrin had been immobilized. These protocols involve protein elution using denaturing buffer solutions. Here we present an improved protocol which permits protein elution under nondenaturing conditions using chelating agents such as phosphate or compounds containing a pyrophosphate group. Furthermore, isothermal titration calorimetry experiments of the purified protein with holotransferrin have been shown to be a reliable method to assess the purity and activity of the purified material. PMID- 11922748 TI - Roles of NADPH-P450 reductase and apo- and holo-cytochrome b5 on xenobiotic oxidations catalyzed by 12 recombinant human cytochrome P450s expressed in membranes of Escherichia coli. AB - Drug oxidation activities of 12 recombinant human cytochrome P450s (P450) coexpressed with human NADPH-P450 reductase (NPR) in bacterial membranes (P450/NPR membranes) were determined and compared with those of other recombinant systems and those of human liver microsomes. Addition of exogenous membrane-bound NPR to the P450/NPR membranes enhanced the catalytic activities of CYP2C8, CYP2C9, CYP2C19, CYP3A4, and CYP3A5. Enhancement of activities of CYP1A1, CYP1A2, CYP1B1, CYP2A6, CYP2B6, CYP2D6, and CYP2E1 in membranes was not observed after the addition of NPR (4 molar excess to each P450). Exogenous purified human cytochrome b5 (b5) further enhanced catalytic activities of CYP2A6, CYP2B6, CYP2C8, CYP2E1, CYP3A4, and CYP3A5/NPR membranes. Catalytic activities of CYP2C9 and CYP2C19 were enhanced by addition of b5 in reconstituted systems but not in the P450/NPR membranes. Apo b5 (devoid of heme) enhanced catalytic activities when added to both membrane and reconstituted systems, except for CYP2E1/NPR membranes and the reconstituted system containing purified CYP2E1 and NPR. Catalytic activities in P450/NPR membranes fortified with b5 were roughly similar to those measured with microsomes of insect cells coexpressing P450 with NPR (and b5) and/or human liver microsomes, based on equivalent P450 contents. These results suggest that interactions of P450 and NPR coexpressed in membranes or mixed in reconstituted systems appear to be different in some human CYP2 family enzymes, possibly due to a conformational role of b5. P450/NPR membrane systems containing b5 are useful models for prediction of the rates for liver microsomal P450-dependent drug oxidations. PMID- 11922749 TI - Soluble expression of a functional recombinant cytolytic immunotoxin in insect cells. AB - We have previously described the production of a recombinant melittin-based cytolytic immunotoxin (IT), scFv-mel-FLAG, in bacterial cells. While the IT exhibited specific cytotoxicity for a human lymphoblastoid cell line, HMy2, yields from expression were low. Here, we describe a baculovirus expression system for the overexpression and secretion of scFv-mel-FLAG. A novel snake phospholipase A2 inhibitor signal peptide was used to aid in the secretion of the immunotoxin. Sf21 insect cells infected with the recombinant virus secreted soluble scFv-mel-FLAG into the culture medium from which it was purified directly on an affinity column. The final yield of scFv-mel-FLAG was estimated at 3-5 mg/L, which was an improvement of 30-fold compared to expression in the prokaryotic system. The cell binding characteristics of the recombinant IT were assessed by flow cytometry using the antigen expressing cell line HMy2. ScFv-mel FLAG bound specifically to HMy2 cells in direct binding assays and this binding was completely inhibited in the presence of an excess of soluble antigen. Significant cytotoxicity for HMy2 cells, measured by leakage of cytosolic LDH, was also observed for the IT at a concentration of 60 pmol/10(4) cells. Cytotoxicity was concentration dependent and was specific for antigen-positive cells. Thus the baculovirus expression system, under the control of a novel secretion signal, can be used for the production of soluble and functional recombinant cytolytic immunotoxins. To our knowledge, this is the first report of expression of a recombinant immunotoxin in the baculovirus expression vector system. PMID- 11922750 TI - Secretory production and purification of functional full-length streptavidin from Bacillus subtilis. AB - Streptavidin is a versatile molecule for many in vitro and in vivo applications. To optimize the production of the full-length streptavidin in a soluble and functional form via secretion using Bacillus subtilis as the expression host, three different strategies were used. These strategies include the construction of a synthetic streptavidin gene, the installation of a transcription terminator, and the use of a sporulation mutant strain. In comparison with the wild-type streptavidin gene in expression studies, a combination of these approaches resulted in a 2.3-fold increase in streptavidin production. The production yields in complex and semidefined media were 94 and 24 mg/liter, respectively. A simple purification scheme which requires only a single ion-exchange matrix was designed to purify streptavidin to homogeneity directly from the culture supernatant. Purified streptavidin was in full length with good biotin binding capacity (3.2 binding sites available per tetramer). A combination of this expression system and purification scheme would be useful for production and purification of high quality functional streptavidin for characterizations and practical applications. PMID- 11922751 TI - Design, production, and characterization of an engineered biotin ligase (BirA) and its application for affinity purification of staphylokinase produced from Bacillus subtilis via secretion. AB - A major attraction in using Bacillus subtilis as an expression host for heterologous protein production is its ability to secrete extracellular proteins into the culture medium. To take full advantage of this system, an efficient method for recovering the target protein is crucial. For secretory proteins which cannot be purified by a simple scheme, in vitro biotinylation using biotin ligase (BirA) offers an effective alternative for their purification. The availability of large amounts of quality BirA can be critical for in vitro biotinylation. We report here the engineering and production of an Escherichia coli BirA and its application in the purification of staphylokinase, a fibrin-specific plasminogen activator, from the culture supernatant of Bacillus subtilis via in vitro biotinylation. BirA was tagged with both a chitin-binding domain and a hexahistidine tail to facilitate both its purification and its removal from the biotinylated sample. We show in this paper how, in a unique way, we solved the problem of protein aggregation in the E. coli BirA production system to achieve a yield of soluble functional BirA hitherto unreported in the literature. Application of this novel BirA to protein purification via in vitro biotinylation in general will also be discussed. Biotinylated staphylokinase produced in the study not only can act as an intermediate for easy purification, it can also serve as an important element in the creation of a blood clot targeting and dissolving agent. PMID- 11922752 TI - Expression in Escherichia coli and purification of hexahistidine-tagged human tissue transglutaminase. AB - Recent evidence suggests that aberrant transglutaminase activity is associated with a wide variety of diseases. Tissue transglutaminase is the most widely distributed of the six well-characterized transglutaminases in humans. We describe a method for expressing hexahistidine-tagged human tissue transglutaminase in Escherichia coli BL21(DE3) using the pET-30 Ek/LIC expression vector. Purification of the expressed enzyme from suspensions of E. coli cells treated with CelLytic B Bacterial Cell Lysis/Extraction Reagent was accomplished by immobilized metal (Ni2+) affinity column chromatography. The procedure typically yields highly purified and highly active recombinant human tissue transglutaminase in about 1 day (about 0.6 mg/from a 1-liter culture). PMID- 11922753 TI - Expression, purification, and isotope labeling of a gp120 V3 peptide and production of a Fab from a HIV-1 neutralizing antibody for NMR studies. AB - Most human immunodeficiency virus type 1 (HIV-1) neutralizing antibodies in infected individuals and in immunized animals are directed against the third variable loop (V3) of the envelope glycoprotein (gp120) of the virus. This loop plays a crucial role in phenotypic determination, cytopathicity (syncytium induction), and coreceptor usage of HIV-1. The human monoclonal antibody 447-52D was found to neutralize a broad spectrum of HIV-1 strains. In order to solve the solution structure of the V3MN peptide bound to the 447-52D Fab fragment by NMR, large quantities of labeled peptide and a protocol for the purification of the Fab fragment were needed. An expression plasmid coding for the 23-residue V3 peptide of the HIV-1MN strain (V3MN peptide, YNKRKRIHIGPGRAFYTTKNIIG) linked to a derivative of the RNA-binding domain of hnRNCP1 was constructed. The fusion protein attached to the V3 peptide prevents its degradation. Using this system, U 15N, U-13C,15N, and U-13C,15N, 50% 2H labeled fusion protein molecules were expressed in Escherichia coli grown on rich Celtone medium with yields of about 240 mg/liter. The V3MN peptide was released by CNBr cleavage and purified by RP HPLC, giving final yields of 6-13 mg/liter. This expression system is generally applicable for biosynthesis of V3-related peptides and was also used to prepare the V3JR-FL. The 447-52D Fab fragment was obtained by a short enzymatic papain cleavage of the whole antibody. Preliminary NMR spectra demonstrate that full structural analysis of the V3MN complexed to the 447-52D Fab is feasible. This system enables studies of the same epitope bound to different HIV-1 neutralizing antibodies. PMID- 11922754 TI - Combined use of regulatory elements within the cDNA to increase the production of a soluble mouse single-chain antibody, scFv, from tobacco cell suspension cultures. AB - In order to facilitate production and secretion of a soluble form of a small, single-chain antibody ScFv (32 kDa) in tobacco cell suspension culture, several modifications were made simultaneously to the antibody cDNA that included elements that have been shown to regulate the expression of proteins in plants. The scFv cDNA was initially ligated into a binary vector under the control of the CaMV 35S promoter and the T7 terminator for expression in tobacco suspension culture. Subsequently, modifications were engineered into the cDNA for enhancement of scFv production. These included the following: (i) the signal peptide (SP) of the tobacco pathogenesis-related protein PR1a which was added in frame to the N-terminal end of scFv cDNA; (ii) a 5'-nontranslated region from the tobacco etch virus (TEV leader sequence), which was fused to the N-terminal end of the SP; and (iii) the endoplasmic reticulum retention signal peptide KDEL, which was added to the C-terminal end of the scFv protein. Using a modified disruption method involving pectinase, the highest expression of total scFv (344 ng scFv/g cell) occurred when the plant leader sequence, the TEV sequence, and the KDEL peptide were all present in the expression construct. Although the addition of the KDEL sequence significantly increased the total yield of protein 5.4-fold, it did not increase the overall amount of protein secreted. These studies indicate that while the SP is very important in promoting secretion of the scFv, it had little influence on increasing scFv secretion levels even when both the TEV and the KDEL sequences significantly increased overall protein levels. PMID- 11922755 TI - Expression and refolding of recombinant human alpha-tocopherol transfer protein capable of specific alpha-tocopherol binding. AB - alpha-Tocopherol transfer protein (alpha-TTP) is a cytosolic protein found predominantly in mammalian liver that is proposed to be responsible for the stereoselective uptake of alpha-tocopherol from the diet. Although recombinant alpha-TTP has been reported previously, little detail has been provided about the yields and competency of the recovered protein at binding tocopherols and other ligands. In this work, we report the successful expression and refolding of a recombinant human alpha-TTP. Ligation-independent cloning generated a construct in pET-30 encoding an alpha-TTP fusion protein (pET-30/ttp) containing a six histidine tag and an S-tag, each cleavable by a separate protease upon expression in Escherichia coli. Overexpression of the protein led to the formation of inclusion bodies that were solubilized in 8 M urea and purified by metal chelate affinity chromatography. Another construct in pET-28b (pET-28b/ttp) provided a soluble protein product after expression that contained a 40-amino-acid N terminal extension, which can be reduced to 21 amino acids by cleavage with thrombin. The success of different refolding experiments was assessed using a Lipidex gel-based tocopherol binding assay. The best recovery of refolded recombinant alpha-TTP fusion capable of binding alpha-tocopherol was provided by matrix-assisted refolding in the presence of 0.5 M arginine. Cleavage of the fusion protein with Factor Xa successfully generated the full-length wild-type protein with no additional N-terminal amino acids. The resulting purification scheme provides recombinant alpha-TTP in good yield and purity for investigation of both its structure and its binding affinities for different ligands including natural and synthetic tocols. PMID- 11922756 TI - Osmoregulatory isoform of dihydroxyacetone phosphate reductase from Dunaliella tertiolecta: purification and characterization. AB - The osmoregulatory isoform of dihydroxyacetone phosphate (DHAP) reductase (Osm DHAPR) is an enzyme unique to Dunaliella, photosynthetic unicellular green algae adapted to extreme environments. This is the first report of purification of an isoform of DHAP reductase from Dunaliella, specifically the osmoregulatory isoform that is involved in the synthesis of free glycerol for osmoregulation in extreme environments, such as high salinity. The Osm-DHAPR is cold labile, inactivated by ammonium sulfate, forms a strong complex with Rubisco, and is unstable in the absence of glycerol. These difficulties have been addressed, and a four-step procedure has been developed to purify the Osm-DHAPR from Dunaliella tertiolecta: precipitation of Rubisco by polyethylene glycol, followed by successive chromatography on DEAE cellulose, Sephacryl S-200, and Red Agarose. Yield of the purified enzyme was 3.6%, with a specific activity of 938 micromol.min-1.mg-1 of protein and a subunit molecular mass of approximately 38 kDa. A maximum specific activity of 2580 micromol.min-1.mg-1 of protein could be achieved by assay with 150 mM NaCl. The Osm-DHAPR had little preference for NADH or NADPH, but it is highly specific for DHAP. Other metabolites of glycolysis, the tricarboxylic acid cycle, and the C3 reductive photosynthetic carbon cycle were not reduced by the enzyme. The purified enzyme was stimulated three-fold by 150 to 250 mM NaCl/KCl and by 25 mM MgCl2. Detergents, lipids, or long-chain acyl CoA derivatives, all of which inhibited the chloroplastic glyceride form of DHAP reductase, did not affect the activity of Osm-DHAPR. The Osm-DHAPR has different properties than the other chloroplastic isoform of DHAP reductase from plants and algae for glycerol phosphate formation and triglyceride synthesis. PMID- 11922757 TI - Utility of (His)6 tag for purification and refolding of proplasmepsin-2 and mutants with altered activation properties. AB - Plasmepsin-2 is a malarial aspartic proteinase that has been implicated in the initial steps of hemoglobin degradation in parasites and thus represents an attractive antimalarial target. Escherichia coli expressed proplasmepsin-2 is capable of activation at acidic pH by autocatalytic cleavage of the pro part region, which results in products of different length. We designed a 10-amino acid deletion in the pro part region that allows faster generation of homogeneous enzyme upon activation. Incorporation of a (His)6 tag onto the N-terminus of the pro part enables on-column refolding of proplasmepsin-2 and simplifies proenzyme purification and pro part separation after activation. The proposed purification procedure results in highly pure and easily crystallizable enzyme. PMID- 11922758 TI - Expression, purification, and structural characterization of human histone H4. AB - Recombinant human histone H4 (hH4) was produced in milligrams quantities in Escherichia coli, without altering the codons of the original cDNA sequence. The hH4 cDNA was subcloned into the pQE30 expression vector, in frame with a sequence encoding an N-terminal stretch of six histidine residues. Purification to electrophoretic homogeneity was obtained by nickel-chelating chromatography, followed by gel filtration. The final yield of the entire expression and purification process was about 1 mg of pure histone H4 per liter of bacterial culture. SDS-PAGE analysis showed for the recombinant H4 a molecular weight corresponding to the expected one (12,535 Da). Circular dichroism spectroscopy was used to estimate the secondary structural composition of recombinant histone, when it is isolated from the physiological core particle. It was observed that under these conditions histone H4 exhibits an altered secondary conformation. In order to induce the recombinant histone to assume a conformation more similar to the one measured when it is organized inside the nucleosome, we resuspended it in buffers at increasing ionic strengths and in the presence of different concentrations of trifluoroethanol. We tried also to mimic the physiological situation of histone H4 by adding an equimolar amount of a commercial DNA to the protein solution. Finally, an estimation of protein thermal stability was evaluated by spectropolarimetry. PMID- 11922759 TI - A flavoprotein encoded in Selenomonas ruminantium is characterized after expression in Escherichia coli. AB - Selenomonas ruminantium is an obligate anaerobe that is very important for the provision of vitamin B12 to ruminants, which are particularly dependent upon this cofactor. One important use for vitamin B12 in anaerobic bacteria is for the utilization of glycerol as carbon source. A new flavoprotein has been found expressed by Escherichia coli from a plasmid created as part of a gene library of S. ruminantium. The 2.5-kb fragment of chromosomal DNA responsible for protein expression contains parts of two operons. Only one polypeptide (the flavoprotein) encoded by the S. ruminantium DNA is produced in E. coli in large amounts. The gene for the flavoprotein has been identified and is probably transcribed as part of an operon involved in glycerol metabolism in S. ruminantium. The flavoprotein has been purified and its molecular properties have been examined. Sequence analysis showed that this protein is a divergent member of the family of nitroreductases. Pure protein is a homodimer with a molecular weight of 44,500, containing one molecule of FMN per dimer. Like other nitroreductases, this protein forms a complex with pyridine nucleotide (NADPH), but unlike other nitroreductases, it fails to be reduced in this complex at a biologically significant rate. It has none of the common catalytic properties of other members of the nitroreductase family. PMID- 11922761 TI - Overexpression, purification, and biochemical characterization of the extracellular human CD83 domain and generation of monoclonal antibodies. AB - CD83 is a 45-kDa glycoprotein and member of the immunoglobulin (Ig) superfamily. It is the best known marker for mature dendritic cells. Although the precise function of CD83 is not known, its selective expression and upregulation together with the costimulators CD80 and CD86 suggests an important role of CD83 in the induction of immune responses. To perform functional studies and to elucidate its mode of action it is vital to obtain recombinant expressed and highly purified CD83 molecules. Therefore, the external Ig domain of human CD83 (hCD83ext) was expressed as a GST fusion protein (GST-hCD83ext) and the soluble protein was purified under native conditions. The fusion protein was purified using GSTrap columns followed by anion-exchange chromatography. GST-hCD83ext was then cleaved using thrombin and soluble hCD83ext was further purified using GSTrap columns and finally by a preparative gel filtration as a polishing step and used for further characterization. The purified GST-hCD83 fusion protein was also used to generate monoclonal anti-CD83 antibodies in a rat system. Two different monoclonal antibodies were generated. Using these antibodies, CD83 was specifically recognized in FACS and Western blot analyses. Furthermore, we showed that native CD83 is glycosylated and that this glycosylation influences the binding of the antibodies in Western blot analyses. Finally, the purified hCD83ext protein was analyzed by one-dimensional NMR and these analyses strongly indicate that hCD83ext is folded and could therefore be used for further structural and functional studies. PMID- 11922760 TI - Tabtoxin-resistant protein: overexpression, purification, and characterization. AB - One of the self-protection mechanisms in Pseudomonas syringae pv. tabaci, a pathogen of tobacco wildfire, is thought to be due to its tabtoxin-resistance gene (ttr). In this study, the ttr gene was inserted into an expression vector, pQE30, and successfully expressed in Escherichia coli M15 at high levels. The purified recombinant tabtoxin-resistant protein (TTR) had an apparent molecular mass of about 21 kDa on SDS-PAGE as well as by mass spectroscopy and had a pI of 6.6 on isoelectric focusing-PAGE. Spectral analysis showed that TTR possesses a maximum fluorescence wavelength (lambda(max)) of 325 nm upon excitation at 282 nm and a positive band with a maximum at 195 nm and a broad negative band with a minimum at 215 nm in the far-UV CD spectrum. The spectrophotometric assay demonstrated the strong detoxification activity of TTR. These results are the first report of the characterization of the purified tabtoxin-resistant protein. Its capacity to detoxify tabtoxinine-beta-lactam shows that it must be one of the self-protection mechanisms in pv. tabaci. PMID- 11922762 TI - High-level expression and secretion of recombinant mouse endostatin by Escherichia coli. AB - The expression of murine endostatin was achieved by placing its gene downstream of an alkaline phosphatase gene (phoA) promoter. To ensure proper folding and secretion of the recombinant protein, the mouse endostatin was fused with alkaline phosphatase signal peptide. SDS/polyacrylamide gel electrophoresis analysis of the culture medium of recombinant Escherichia coli cells revealed that endostatin was efficiently secreted. The signal peptide was efficiently cleaved during secretion as demonstrated by N-terminal amino acid sequencing. The maximum yield of secreted endostatin during fermentation was 40 mg/liter. Up to 28 mg of endostatin was purified from 1 liter of cell culture broth. The biological activity of recombinant protein was tested in a bovine aortic endothelial (BAE) cell proliferation assay. The recombinant endostatin inhibited the growth of BAE cells stimulated by basic fibroblast growth factor, and its ED50 was comparable to that from a previous report. Flow cytometric measurements of BAE cells cultivated in medium with endostatin demonstrated a cell cycle arrest mainly in the G0/G1 phase and a decrease in the S phase. PMID- 11922763 TI - Affinity extraction of dye- and metal ion-binding proteins in polyvinylpyrrolidone-based aqueous two-phase system. AB - Affinity extraction of dye- and metal ion-binding proteins, respectively, in a polyvinylpyrrolidone (PVP40)-Reppal PES 100 two-phase system was investigated. Due to the ability of PVP to complex azo dyes and inorganic ions, covalent coupling of the ligands was not essential. Cibacron Blue F3GA was used as the ligand for extraction of lactate dehydrogenase (LDH) from porcine muscle, while copper ions were used for extraction of B. stearothermophilus LDH with a fusion tag of six histidine residues (His6-LDH) from recombinant Escherichia coli homogenate. The binding strength of the enzymes to their respective ligands was only slightly reduced in the presence of PVP. The partition coefficient of Cibacron Blue and Cu2+ ions in the two-phase systems composed of different concentrations of PVP and Reppal was in the range of 20-30, with maximal partitioning being observed in the 17% (w/w) PVP40-10% Reppal PES100 system. Only a minor leakage of the ligands to the bottom phase was observed with time. The partitioning of porcine LDH to the PVP phase was increased 100-fold, and a maximal recovery of 89% was obtained in the two-phase system loaded with 0.2% (w/w) Cibacron Blue. The enzyme was quantitatively recovered with further purification from the PVP-dye phase using a secondary extraction step with 170 mM phosphate or alternatively with 100 mM phosphate containing NADH or NaCl. A more than 10-fold increase in the partition coefficient of His6-LDH was achieved in the two-phase system loaded with 0.4% (w/w) copper sulfate compared to the system lacking the metal ions. The enzyme was also back-extracted into phosphate phase in the presence of imidazole. PMID- 11922764 TI - Effect of tandem rare codon substitution and vector-host combinations on the expression of the EBV gp110 C-terminal domain in Escherichia coli. AB - Gp110 of Epstein-Barr virus (EBV) is a glycoprotein that functions exclusively during the assembly of EBV nucleocapsid and the release of infectious EBV. Its C terminal tail domain (gp110 CTD) is essential for gp110's function and may provide signals that are responsible for the assembly and release of EBV. In the present study, to get large amounts of gp110 CTD for structural analysis, the effects of vector system, codon usage, and host strain on expression levels of gp110 CTD in Escherichia coli have been investigated. The coding region of gp110 CTD (11 kDa) was subcloned into the expression vectors pSE 280, pET-15b, pET-29a, pMAL-c2x, and pGEX-4T-1. Except the pMAL-c2x construct, all the others failed to express detectable amounts of recombinant gp110 CTD. Substituting a tandem rare AGA (Arg) codon with a synonymous CGC (Arg) codon facilitated expression of the recombinant protein, while a protease-deficient host E. coli strain helped in the accumulation of a soluble form of gp110 CTD fusion. The secondary structures of the obtained recombinant gp110 CTD purified from soluble extracts and inclusion bodies were compared using circular dichroism analysis. In aqueous solutions, both samples equally adopt a mixed alpha-helix and beta-sheet conformation as well as a partly unordered structure. Notably, in the membrane-mimicking environments the helical propensity of gp110 CTD increased up to the previously predicted level based on its sequence, suggesting that gp110 CTD may fold into a more stable conformation through interactions with the cell membrane. PMID- 11922765 TI - A mammalian expression system for rapid production and purification of active MAP kinase phosphatases. AB - Expression of enzymatically active mammalian proteins in Escherichia coli can proven to be a challenging task due to poor solubility, improper folding, and lack of adequate posttranslational modification. Expression of mammalian proteins using baculovirus or yeast systems is time-consuming and may also be subject to inadequate modification. In order to overcome these technical difficulties, we have developed a mammalian expression system for the convenient subcloning of cDNA fragments, high-level expression, and one-step purification of enzymatically active proteins. The mammalian expression vector pEBG that expresses glutathione S-transferase fusion proteins was modified to create an SrfI restriction site in the multiple cloning site. The protein coding sequences of MAP kinase phosphatase 1 (MKP-1), MAP kinase phosphatase-2 (MKP-2), and the tumor suppressor PTEN were PCR-amplified using Pfu DNA polymerase and cloned into the SrfI site through SrfI digestion-coupled ligation. The resulting plasmids were transiently transfected into 293T cells using FuGENE 6 transfection reagent. Forty eight hours after transfection, cells were harvested and bioactive recombinant proteins were purified by glutathione-Sepharose beads. Protein yield, which ranged from 200 to 700 microg, was more than adequate for biochemical studies. The usefulness of this versatile system for studying protein function and its potential application for proteomics research are discussed. PMID- 11922766 TI - Purification and characterization of the major lipoprotein (P28) of Spiroplasma apis. AB - The plasma membrane of Spiroplasma apis contains a 28-kDa major protein (P28), like other spiroplasmas which also possess a main 26- to 28-kDa membrane polypeptide, called spiralin. In the work described here, we have developed a simple and efficient method for the purification of P28 of this mollicute, a wall less eubacteria. Proteins were first selectively extracted from the isolated membrane with the mild detergents (i) sodium N-lauroylsarcosinate (Sarkosyl) and (ii) 3-[(3-cholamidopropyl)dimethylamonio]-1-propyl sulfonate (Chaps) and subjected to size-exclusion HPLC in the presence of Chaps. The P28-enriched fraction was thereafter subjected to the second chromatographic step involving cation exchange HPLC in the presence of the same detergent. P28 was purified at the milligram level (yield, 40%). Metabolite labeling with [14C]palmitic acid and chemical analysis of P28 indicated that it is covalently modified by two O-ester bound fatty acids and one amide-linked chain and contains a S-glycerylcysteine at the N-terminus. By charge-shift electrophoresis, Triton X-114 phase separation, and growth inhibition tests it was shown that P28 is a typical amphiphilic protein exposed, at least partly, at the cell surface. Together, our data provided evidence that P28 is a "classical" lipoprotein (i.e., triacylated) like the members of the spiralin family. PMID- 11922767 TI - Expression and purification of homogenous proteins in Saccharomyces cerevisiae based on ubiquitin-FLAG fusion. AB - The construction of an expression vector for increased expression of cytoplasmic proteins in Saccharomyces cerevisiae is described. To enhance the yield of expressed proteins, fusion of ubiquitin to an octapeptide (a FLAG tag) upstream of the respective model genes was applied. During protein maturation ubiquitin is efficiently removed by yeast autologous hydrolases, generating the FLAG octapeptide at the N-terminus. Fusion proteins were recognized by the specific monoclonal antibody M1 directed against the FLAG tag. The FLAG-tagged proteins were purified to homogeneity by immunoaffinity chromatography using an anti-FLAG M1 agarose. Different model proteins, green fluorescent protein, green fluorescent protein-human lysozyme, green fluorescent protein elongation initiahon factor 5a, green fluorescent protein-rapamycin-selective 25-kDa immunophilin, and green fluorescent protein-heat shock protein 90 beta have been selected to demonstrate the efficiency of the new vector construct. PMID- 11922768 TI - Characterization of an antibody Fv fragment that binds to the human, but not to the rat neurotensin receptor NTS-1. AB - The cDNAs coding for the heavy and light chain variable domains of an antibody, recognizing the human G-protein-coupled receptor for neurotensin, NTS-1, were obtained from a hybridoma cell line, B-N6. The Fv B-N6 fragment was expressed in Escherichia coli and purified. To characterize the properties of the antibody fragment, human and rat high-affinity neurotensin receptors were expressed in E. coli in functional form, linked at their N-termini to the maltose-binding protein. Fv B-N6 was found to compete for [3H]neurotensin binding to the human neurotensin receptor, but not to the rat neurotensin receptor, with IC50 values of 1.6 microM (membrane-bound receptor) and 1.9 microM (detergent-solubilized, purified receptor). The formation of a relatively stable complex of Fv B-N6 with purified human neurotensin receptor fusion protein was also demonstrated by gel filtration experiments. The Fv B-N6 fragment will be used to isolate a high affinity binder to the human neurotensin receptor as a valuable tool for cocrystallization and receptor structure determination. PMID- 11922769 TI - Expression of FLAG fusion proteins in insect cells: application to the multi subunit transcription/DNA repair factor TFIIH. AB - The multi-subunit transcription/DNA repair factor TFIIH was used as a model system to show that the expression of FLAG fusion proteins in insect cells constitutes a versatile tool for both structural and functional investigations. In the present study, we have constructed recombinant baculoviruses expressing the four core TFIIH subunits fused at their N-terminus to the FLAG peptide. Using these recombinant viruses we have established protocols based on anti-FLAG immunoaffinity chromatography that allow the systematic analysis of pairwise interaction within multiprotein complexes and have developed a double tag strategy (FLAG and hexahistidine tags) for the identification and purification of stable TFIIH subcomplexes. A simple purification procedure was developed that leads to the isolation of recombinant TFIIH containing the full set of subunits. The purified recombinant TFIIH was shown to be active in a transcription assay and to be structurally homologous to the endogenous complex by electron microscopy and image analysis. PMID- 11922770 TI - Delandin, a chitinase-like protein with antifungal, HIV-1 reverse transcriptase inhibitory and mitogenic activities from the rice bean Delandia umbellata. AB - An antifungal protein with a chitinase-like N-terminal sequence, designated delandin, was isolated from the rice bean. The protein exhibited a molecular weight of 28 kDa and was adsorbed on both blue Affi-Gel and SP-Toyopearl. It exerted antifungal action toward Mycosphaerella arachidicola, Botrytis cinerea, Fu- sarium oxysporum, Rhizoctonia solani, and Colletotrichum gossypii and inhibited the activity of human immunodeficiency virus 1 reverse transcriptase. The protein inhibited translation in rabbit reticulocyte lysate with a low potency. It elicited a mitogenic response from mouse splenocytes. PMID- 11922771 TI - An X-prolyl dipeptidyl aminopeptidase from Lactococcus lactis: cloning, expression in Escherichia coli, and application for removal of N-terminal Pro-Pro from recombinant proteins. AB - A novel pepX gene was cloned from isolated DNA of Lactococcus lactis by PCR. The deduced amino acid sequence of the 89-kDa protein showed 94, 93, 65, and 44% identity with the pepX protein from Lactococcus lactis subsp. cremoris, Lactococcus lactis subsp. lactis, Lactobacillus delbruecki subsp. bulgaricus, and Lactobacillus helveticus, respectively, and contained a serine protease G-K-S-Y-L G consensus motif. The pepX gene has been cloned into pET17b and was expressed at a high level in Escherichia coli BL21 (DE3) LysS. PepX was purified to approximate homogeneity with ammonium sulfate precipitation and DEAE Sephadex A 50 chromatography. Optimal pepX activity was observed at pH 8.0 and 37 degrees C. According to SDS-PAGE analysis, pepX has a molecular mass of approximately 89 kDa. The peptidase can remove completely the unwanted X-Pro from the N-terminal of the target protein, releasing the naturally active protein and peptide, revealing a prospective application of pepX in large-scale production of pharmaceutical protein and peptide products. PMID- 11922772 TI - Estrogen-dependent regulation of the expression of hepatic Cyp2b and 3a isoforms: assessment using aromatase-deficient mice. AB - The role of estrogen in the expression and induction of hepatic Cyp2b and Cyp3a isoforms was studied using mice [Ar (-/-) mice] lacking aromatase, a key enzyme for estrogen biosynthesis. The expression of P450s was determined by reverse transcription-polymerase chain reaction, immunoblotting, and measuring testosterone 6beta- and 16alpha-hydroxylase activity as markers. Basic expression of Cyp3a11 mRNA and protein was seen in both sexes of Ar (+/+) mice. Disruption of the aromatase gene caused an increase in the expression of Cyp3a11 protein, although the mRNA level remained unchanged. Female-specific Cyp3a41 disappeared in Ar (-/-) mice, and this could not be reversed by administration of exogenous beta-estradiol to adult knockout mice. The constitutive expression of female specific Cyp2b9 also disappeared on disrupting the aromatase gene. However, in clear contrast to Cyp3a41, some individual Ar (-/-) mice exhibited expression of this form following treatment with exogenous beta-estradiol. Disruption of the aromatase gene had no effect on PB-mediated induction of Cyp2b10 or on the noninducible nature of Cyp2b9, Cyp3a11, and Cyp3a41. These results suggest that (1) Cyp3a11 is suppressed by estrogen; (2) the expression of female-specific Cyp3a41 is programmed by neonatal and/or infantile exposure to estrogen; (3) maintenance of the expression of female-specific Cyp2b9 requires estrogen in adults; and (4) endogenous estrogen plays little, if any, role in the mechanism by which PB induces Cyp2b10. PMID- 11922773 TI - Assessment of interactions of diverse ternary mixtures in an estrogen receptor alpha reporter assay. AB - This study used an MCF-7 cell based ER-alpha reporter gene assay to assess chemical interactions within the following ternary mixtures: (1) three synthetic pesticides, methoxychlor (MXC), o,p-DDT, and dieldrin; (2) three polyaromatic hydrocarbons, benzo[a]pyrene (BAP), 1,2-benzanthracene (BENZ), and chrysene (CHRY); and (3) an endogenous estrogen, [17beta-estradiol, (E(2))]; a phytoestrogen, genistein (GEN); and a synthetic estrogen, o,p-DDT. A full factorial design in which four concentrations of each chemical were assessed in all possible combinations (64 treatment groups) was utilized. In addition, mixtures were tested in both a low range (concentrations near the individual chemical response thresholds) and a high range ( approximately 2-10x higher) experiment. A response surface was estimated using a nonlinear mixed model, and the cumulative response in each mixture was evaluated for departure from additivity. The mixture of E(2), GEN, and DDT exhibited antagonistic interactions (p < 0.001) in both concentration ranges. However, specific interactions between E(2)/GEN and E(2)/DDT differed between the low and high range concentrations. The BAP/BENZ/CHRY mixture did not depart significantly from additivity (p = 0.66) in either concentration range, although response levels were generally low. The MXC/DDT/dieldrin mixture did not depart significantly from additivity in either the high (p = 0.065), or low dose range (p = 0.506), with generally minimal responses dominated by MXC and DDT. This methodology has allowed for a rigorous statistical evaluation of potential departures from additive interactions in endocrine active mixtures. In no case was a significantly greater-than-additive (synergistic) interaction observed. PMID- 11922774 TI - Antitumor effects of miconazole on human colon carcinoma xenografts in nude mice through induction of apoptosis and G0/G1 cell cycle arrest. AB - Miconazole (MIC), a promising oral antifungal agent, has been used worldwide in the treatment of superficial mycosis. In this study, we demonstrated that MIC dose dependently arrested various human cancer cells at the G0/G1 phase of the cell cycle. The protein levels of p53, p21/Cip1, and p27/Kip1 were significantly elevated by MIC treatment in COLO 205 cells. Electrophoretic mobility gel shift assays showed that the nuclear extracts of the MIC-treated COLO 205 cells exerted a significant binding between wild-type p53 and its consensus-binding site present in the p21/Cip1 promoter. These results suggested that the p53-associated signaling pathway is involved in the regulation of MIC-induced cancer cell growth arrest. By immunoblot analysis, we demonstrated that cyclin D3 and cyclin dependent kinase-4 (CDK4) protein levels were inhibited by MIC treatment in the cancer cells. Significant therapeutic effect was further demonstrated in vivo by treating nude mice bearing COLO 205 tumor xenografts with MIC (50 mg/kg ip). The protein expression of p53 was significantly increased in MIC-treated tumor tissues by immunohistochemical staining and Western blotting analysis. DNA fragmentation and TUNEL assay were performed and demonstrated that apoptosis occurred in tumor tissues treated with MIC. Our study provides the novel mechanisms of antitumor effects of MIC and such results may have significant applications for cancer chemotherapy. PMID- 11922775 TI - Interferon-alpha2b secretion by adenovirus-mediated gene delivery in rat, rabbit, and chimpanzee results in similar pharmacokinetic profiles. AB - Gene delivery, with subsequent protein synthesis and secretion, in vivo has been proposed as an alternative way to deliver a therapeutic protein to the systemic circulation. Interferon-alpha (IFN) protein is effective in the treatment of viral and malignant diseases but has short serum half-life that requires frequent administration. An E1 region-deleted adenovirus vector encoding human IFN-alpha2b gene driven by the cytomegalovirus immediate early promoter (rAd-IFN) was generated to assess the serum concentration-time profiles of expressed IFN protein in animal models. Intravenous administration of rAd-IFN, normalized for body weight, resulted in dose-dependent serum IFN concentrations that persisted 8 40 days with similar concentration-time profiles in rats and rabbits. We sought to determine if serum concentration-time profiles in the rat and rabbit animal models would be predictive for a larger animal and would therefore be relevant models for potential dosing of human patients. Two chimpanzees (approximately 70 kg) dosed with rAd-IFN by intravenous administration normalized to body weight achieved serum IFN concentration-time profiles similar to those observed in rats and rabbits. The role of the immune response in limiting the persistence of transgene expression was highlighted by the persistence of serum IFN concentrations for over 200 days in beige/SCID immunodeficient mice. These studies suggest that serum concentration of secreted transgene products after gene delivery in small animal models may be highly predictive for larger species and will help define dosing strategies in human patients. PMID- 11922776 TI - Endotoxin potentiation of trichothecene-induced lymphocyte apoptosis is mediated by up-regulation of glucocorticoids. AB - Exposure to bacterial endotoxin (lipopolysaccharide, LPS) is quite common and may increase human susceptibility to chemical-induced tissue injury. The purpose of this study was to identify mechanisms by which LPS potentiates lymphoid tissue depletion in B6C3F1 mice exposed to the common food-borne trichothecene mycotoxin, vomitoxin (VT). As demonstrated by DNA fragmentation and flow cytometric analysis, apoptosis in thymus, Peyer's patches, and bone marrow was marked in mice 12 h after administering Escherichia coli LPS (0.1 mg/kg body wt ip) concurrently with VT (12.5 mg/kg body wt po), whereas apoptosis in control mice or mice treated with either toxin alone was minimal. Based on observed increases in tumor necrosis factor-alpha (TNF-alpha) and interleukin (IL)-6 serum concentrations following LPS and VT cotreatment, the roles of these cytokines in apoptosis potentiation were assessed. Injection with rolipram, an inhibitor of TNF-alpha expression, or use of IL-6 knockout mice was ineffective at impairing thymic apoptosis induction by the toxin cotreatment, suggesting that these cytokines did not mediate LPS potentiation. Toxin cotreatment increased splenic cyclooxygenase-2 mRNA expression, suggesting possible involvement of prostaglandins in apoptosis. However, indomethacin, a broad spectrum inhibitor of cyclooxygenases, failed to block thymus apoptosis. Toxin cotreatment increased serum corticosterone and, furthermore, RU 486, a glucocorticoid receptor antagonist, significantly abrogated apoptosis in thymus, Peyer's patches, and bone marrow following LPS + VT exposure. The results presented herein and the known capacity of glucocorticoids to cause apoptosis indicate that hypothalamic pituitary-adrenal axis plays a key role in LPS potentiation of trichothecene induced lymphocyte apoptosis. PMID- 11922777 TI - Role of rat multidrug resistance protein 2 in plasma and biliary disposition of dibromosulfophthalein after microsomal enzyme induction. AB - We have previously demonstrated that microsomal enzyme inducers phenobarbital (PB) and pregnenolone-16alpha-carbonitrile (PCN), but not 3-methylcholanthrene (3 MC) and benzo(a)pyrene (BaP), increase expression and function of rat Multidrug Resistance Protein 2 (Mrp2), a canalicular organic anion transporter. Thus, the purpose of this study was to determine whether Mrp2 protein induction alters the biliary and plasma dispositions of dibromosulfophthalein (DBSP). After four daily ip injections of PB, PCN, 3-MC, BaP, or vehicle, DBSP (100 mg/kg) was injected iv and was measured in blood and bile over a 40-min period. PB and PCN significantly enhanced plasma disappearance and biliary excretion of DBSP, whereas 3-MC and BaP did not. To determine whether the enhanced plasma disappearance and biliary excretion was entirely due an increase in Mrp2, PCN was also administered ip daily for 4 days to Mrp2-null Eisai hyperbilirubinemic (EHBR) rats and then injected iv with DBSP. PCN significantly increased plasma DBSP disappearance in EHBR rats during early time intervals (2-20 min), but not at later time intervals (25-40 min). PCN did not increase DBSP biliary excretion in EHBR rats, but actually decreased it at later time intervals. In summary, the increase in Mrp2 protein after microsomal enzyme induction is responsible for increased biliary DBSP excretion. Furthermore, the increase in Mrp2 protein after microsomal enzyme induction is not responsible for the enhanced plasma DBSP disappearance at early time points, yet may influence plasma DBSP disappearance at later time points. This study also demonstrates the importance of compensatory hepatic transporters in eliminating DBSP by alternative pathways other than Mrp2. PMID- 11922778 TI - Scientific significance on the basis of the quality of mitochondrial preparations with which the experiments were performed. PMID- 11922780 TI - Diastereoselective synthesis of psi[(E)-CH=CMe]- and psi[(Z)-CH=CMe]-type dipeptide isosteres by organocopper-mediated anti-S(N)2' reaction. AB - [reaction: see text] Acyclic psi[(E)-CH=CMe]- and psi[(Z)-CH=CMe]-type dipeptide isosteres were efficiently synthesized. In a key reaction, alpha-alkylation of gamma-mesyloxy-beta-methyl-alpha,beta-unsaturated esters with organocyanocuprates in diethyl ether or tetrahydrofuran preferentially afforded the psi[(E)-CH=CMe]- or psi[(Z)-CH=CMe]-isomer, respectively, via anti-S(N)2' mechanism. PMID- 11922781 TI - Diastereoselective synthesis of psi[(E)-CMe=CH]- and psi[(E)-CMe=CMe]- type dipeptide isosteres based on organocopper-mediated anti-S(N)2' reaction. AB - [reaction: see text] A straightforward synthetic route for the synthesis of diastereomerically pure psi[(E)-CMe=CH]- and psi[(E)-CMe=CMe]-type dipeptide isosteres was developed on the basis of regio- and stereoselective anti-S(N)2' alkylation of 3-(N-Boc-5-methyl-4-substituted-oxazolidin-2-on-5-yl)acrylates with organocopper reagents. PMID- 11922782 TI - The effect of long-range circumannular non-bonding orbital interaction on the inversion of groups on nitrogen in 2,2,4,4-tetramethyl-3-imino-1-cyclobutanones. AB - [structure: see text] The free energy of activation, deltaG*, for nitrogen inversion was calculated from the coalescence temperature for the methyl signals obtained via variable temperature NMR for a series of imino derivatives of the title compounds. There was a direct correlation between the long-range nonbonding circumannular orbital interactions between the heteroatoms in the 1,3-positions and the deltaG* for inversion of groups on nitrogen. Values of deltaG* range from 26 kcal/mol on systems with maximum interaction in the transition state (I and II) to 17 kcal/mole on those systems with minimal interaction (III and IV). PMID- 11922783 TI - Studies toward the total synthesis of diterpene antibiotic guanacastepene A: construction of the hydroazulenic core. AB - [reaction: see text] As a part of studies aimed toward the total synthesis of biologically important natural product guanacastepene A of contemporary interest, a new and concise route to a fully functionally endowed hydroazulenic core is delineated. The strategy involves the building of the requisite stereochemical features on a endo-tricyclo[5.2.1.0(2,6)]decane matrix and excision of the five membered ring through a retro-Diels-Alder reaction. Generation of the seven membered ring to access the hydroazulenic framework was achieved employing ring closure metathesis (RCM) reaction as the key step. PMID- 11922784 TI - 4,5-diazafluorene-based overcrowded alkene: a new ligand for transition metal complexes. AB - [structure: see text] A new ligand system, where a 4,5-diazafluorene-type chelate and a methoxybenzoxanthene unit are coupled by a double bond has been synthesized and fully characterized including X-ray structure. The synthesis and UV-vis spectra of Ru(II), Os(II), and Re(I) complexes with the above-mentioned ligand are also shown. PMID- 11922785 TI - Enhanced regioselectivity of Yang photocyclization in the crystalline state. AB - [reaction: see text] In contrast to the solution state, where Yang photocyclization of ketones of general structure 1 leads to equal amounts of cyclobutanols 2 and 3 (by abstraction of HA and HB, respectively), irradiation in the crystalline state is much more regioselective, favoring either 2 or 3 depending on the nature of the substituent X. X-ray crystallography and molecular mechanics calculations reveal the source of this remarkable solid-state regioselectivity. PMID- 11922786 TI - Facile, efficient conjugation of a trifunctional lanthanide chelate to a peripheral benzodiazepine receptor ligand. AB - [reaction: see text] Receptor-mediated imaging and therapy of diseased tissue is rapidly gaining favor in the medical community. The synthesis and facile aqueous/organic coupling of a peripheral-type benzodiazepine receptor ligand to a cyclen-based fluorophore is described herein. The contrast agent QM-CTMC-PK11195, when chelated with lanthanides, produces bright luminescence and good MRI contrast and can potentially serve as an imaging and demarcation agent for certain types of cancers. PMID- 11922787 TI - Deglycobleomycin: solid-phase synthesis and DNA cleavage by the resin-bound ligand. AB - [structure: see text] A greatly improved solid-phase synthesis of deglycobleomycin using a Dde-based linker is reported. The resin-bound deglycobleomycin could be completely deblocked and assayed for DNA plasmid relaxation, sequence-selective DNA cleavage, and light production from a molecular beacon. PMID- 11922788 TI - Total synthesis of cis-solamin. AB - [structure: see text] A convergent total synthesis of cis-solamin and its diastereomer was accomplished using VO(acac)2-catalyzed diastereoselective epoxidation followed by cyclization of bis-homoallylic alcohol as the key step. By comparison of the optical rotation of two possible diastereomers, it is suggested that the absolute configuration of natural cis-solamin is 1a. PMID- 11922789 TI - Total syntheses of (-)-fumiquinazolines C, E, and H. AB - [structure: see text] Total syntheses of the heptacyclic fumiquinazolines C and H have been accomplished efficiently using FmocNHCH(CH2SePh)CO2H as the precursor for the requisite dehydrofumiquinazoline. PMID- 11922790 TI - Nazarov reagents for convergent and expedient synthesis of new steroids. AB - [reaction: see text] A new methodology for the convergent synthesis of tetracyclic compounds was developed. Two new bicyclic Nazarov reagents of type II were synthesized, and their cycloaddition with 2-carbomethoxy-2-cyclohexenone I was studied. This cycloaddition afforded new interesting steroidal backbones. PMID- 11922791 TI - Somocystinamide A, a novel cytotoxic disulfide dimer from a Fijian marine cyanobacterial mixed assemblage. AB - [reaction: see text] Bioassay-guided investigation of the extract from a Lyngbya majuscula/Schizothrix sp. assemblage of marine cyanobacteria led to the discovery of somocystinamide A (1), an extraordinary disulfide dimer of mixed PKS/NRPS biosynthetic origin. Somocystinamide A (1) was highly acid-sensitive, rapidly and completely converting to a characterizable derivative (2). Compound 1 exhibits significant cytotoxicity against mouse neuro-2a neuroblastoma cells (IC50 = 1.4 microg/mL), whereas 2 has no activity. PMID- 11922792 TI - A novel photochromic system of 4,5-dialkenylthiophenes constructed by the smarium diiodide promoted coupling reactions of thiophene-2-carboxylate with aryl ketones. AB - [reaction: see text] The SmI2-promoted coupling reaction of ethyl thiophene-2 carboxylate with aryl ketones (2 equiv), followed by acid-catalyzed dehydration and oxidative aromatization, gave dialkenylthiophenes 1b-d, which underwent electrocyclizations upon irradiation with 300-nm light in CH3CN solution to give the corresponding closed-ring species with absorption lambda(max) approximately 425 nm. The interconversion between dialkenylthiophenes and their corresponding closed-ring species constitutes a novel photochromic system bearing an ester group for potential uses in linkage and wavelength tuning. PMID- 11922793 TI - A strategy for the synthesis of aryl alpha-ketoamides based upon the acylation of anions derived from cyanomethylamines followed by oxidative cleavage. AB - [reaction: see text] Cyanomethylamines, prepared by alkylation of amines with chloroacetonitrile, were deprotonated using NaHDMS in THF, reacted with heteroaryl or substitutedphenyl esters, and then oxidized by adding Clorox(TM) to afford aryl alpha-ketoamides in a single operation in good overall yields. PMID- 11922794 TI - Enantioselective aziridination of alkenes with N-aminophthalimide in the presence of lead tetraacetate-mediated chiral ligand. AB - [reaction: see text] Reaction of various N-enoyl oxazolidinones 5a-f with N aminophthalimide and lead tetraacetate in the presence of camphor-derived chiral ligands provides the desired N-phthalimidoaziridines 6a-f in good to high enantiomeric excess (67-95% ee) at 0 degrees C within 15 min. The absolute stereochemistry of the corresponding aziridine derivatives was established by chemical correlations. PMID- 11922795 TI - Nucleophilic addition reaction of 2-trimethylsilyloxyfuran to N-gulosyl-C alkoxymethylnitrones: synthetic approach to polyoxin C. AB - [reaction: see text] The stereoselectivity of nucleophilic addition of 2 trimethylsilyloxyfuran to N-gulosyl-C-alkoxymethylnitrones was investigated. It was found that the selectivity was highly dependent on the bulkiness of the C substituent of the nitrone. The major adducts were elaborated into the key intermediate of polyoxin C. PMID- 11922796 TI - 1-dodecyloxy-4-perfluoroalkylbenzene as a novel efficient additive in aldol reactions and Friedel-Crafts alkylation in supercritical carbon dioxide. AB - [reaction: see text] 1-Dodecyloxy-4-perfluoroalkylbenzenes accelerated organic reactions such as aldol-type reactions and Friedel-Crafts alkylation in supercritical carbon dioxide. The perfluoroalkylbenzene worked as a surfactant in the system, and formation of emulsions during the reactions was observed. PMID- 11922797 TI - Tuning rate of the bergman cyclization of benzannelated enediynes with ortho substituents. AB - [reaction: see text] The Bergman cyclization of benzannelated enediynes is highly sensitive to ortho substitution. This finding opens possibilities for the rational design of conformer-specific and pH-dependent DNA-cleaving agents. PMID- 11922798 TI - Structure, stereochemistry, and biological activity of integramycin, a novel hexacyclic natural product produced by Actinoplanes sp. that inhibits HIV-1 integrase. AB - [structure: see text] HIV-1 integrase is a critical enzyme for viral replication, and its inhibition is an emerging target for potential antiviral chemotherapy. We have discovered a novel inhibitor, integramycin, from screening of fermentation extracts using an in vitro assay. Integramycin possesses a hexacyclic ring system and exhibited an IC50 value of 4 microM against HIV-1 integrase (strand transfer). The isolation, structure elucidation, stereochemistry, conformation, and biological activity has been described. PMID- 11922799 TI - Magnesium halide-catalyzed anti-aldol reactions of chiral N acylthiazolidinethiones. AB - [reaction: see text] Diastereoselective direct aldol reactions of chiral N acylthiazolidinethiones occur in high yield with preference for the illustrated anti diastereomer. This reaction is catalyzed by 10% MgBr2.OEt2 in the presence of triethylamine and chlorotrimethylsilane. Yields range from 56 to 93% with diastereoselectivity up to 19:1 for a variety of N-acylthiazolidinethiones and unsaturated aldehydes. PMID- 11922800 TI - p-Methoxybenzyl ether cleavage by polymer-supported sulfonamides. AB - [reaction: see text] p-Methoxybenzyl ethers have been found to transfer from alcohols to sulfonamides in the presence of catalytic trifluoromethanesulfonic acid. This process for protecting group removal can be performed in solution with yields >94%. Through the use of sulfonamide-functionalized ("safety-catch") resins, p-methoxybenzyl ethers can be cleaved in excellent yields with minimal purification. PMID- 11922801 TI - A short synthesis of the erythrina skeleton and of (+/-)-alpha-lycorane. AB - [reaction: see text] A new nonchain 5-endo radical cyclization starting with xanthates was exploited in a short synthesis of (+/-)-alpha-lycorane and the erythrina ring system. PMID- 11922802 TI - Synthesis of 1,2-dithiolane analogues of leucine for potential use in peptide chemistry. AB - [reaction: see text]1,2-Dithiolanes present several points of interest for both peptide and medicinal chemistry, yet no chiral alpha-amino acids containing this five-membered heterocyclic system are available. We report here the first synthesis of N- and C-protected derivative of (S)-2-amino-3-(1,2-dithiolan-4 yl)propionic acid (Adp) and its 1,3-dithiolic form. PMID- 11922803 TI - Palladium-catalyzed tandem cyclization of allenyl-aldehydes and -ketones with aryl iodides and Bu3SnSnBu3. AB - [reaction: see text] The arylative cyclization of allenyl-aldehydes with aryl iodides and hexa-n-butyldistannane to form substituted cyclopentanols and cyclohexanols by tandem palladium-catalyzed carbostannylation and allylation was achieved under mild conditions. PMID- 11922804 TI - Synthesis of L-oxetanocin. AB - [reaction: see text] Hitherto unknown L-oxetanocin has been synthesized from L xylose in 16 steps via a ribonolactone derivative. PMID- 11922806 TI - The diverted di-pi-methane rearrangement; mechanistic and exploratory organic photochemistry. AB - [reaction: see text] An unusual diversion of the di-pi-methane rearrangement has been encountered. The reaction is characteristic of di-pi-methane systems having one vinyl group bearing one or two carbonyl groups and provides a synthesis of dihydrofurans. PMID- 11922805 TI - Concise synthesis of anhydrovinblastine from leurosine. AB - [reaction: see text] The Cp(2)TiCl-mediated deoxygenation of leurosine (1) afforded anhydrovinblastine (4) in good yield. Furthermore, as the reaction proceeded via a carbon-centered radical intermediate, this transient was also trapped by a hydrogen-atom donor to afford selectively reduced alkaloid 10. PMID- 11922807 TI - A short synthesis of (+/-)-epiasarinin. AB - [reaction: see text] Epiasarinin, an endo-endo furofuran, has been synthesized from piperonal via a five-step route with good stereocontrol. The sequence involves Darzens condensation, alkenyl epoxide-dihydrofuran rearrangement, and a Lewis acid mediated cyclization. PMID- 11922808 TI - Broadly effective enantioselective Diels-Alder reactions of 1-amino-substituted 1,3-butadienes. AB - [reaction: see text] A broad range of substituted 1-amino-1,3-butadienes undergo enantioselective Diels-Alder reactions with methacrolein in the presence of 5 mol % of Cr(III)-salen complex 1. The reactions are carried out conveniently, at room temperature, and they afford the cycloadducts in high yields and excellent ee's. PMID- 11922809 TI - A new resin-bound universal isonitrile for the Ugi 4CC reaction: preparation and applications to the synthesis of 2,5-diketopiperazines and 1,4-benzodiazepine-2,5 diones. AB - [reaction: see text] The preparation and synthetic applications of a novel resin bound isonitrile are described. The resin is an example of a novel convertible isonitrile that can be utilized in the Ugi multicomponent reaction. Base activation of the resin-bound Ugi product results in cleavage via formation of a N-acyloxazolidone that is then trapped as a carboxylic acid ester. This resin and the methodology described are suitable for synthesizing diversity libraries of 2,5-diketopiperazines and 1,4-benzodiazepine-2,5-diones. PMID- 11922810 TI - Regioselective synthesis of 3-alkynyl-5-bromo-2-pyrones via Pd-catalyzed couplings on 3,5-dibromo-2-pyrone. AB - [reaction: see text] 3,5-Dibromo-2-pyrone underwent facile Pd(0)-catalyzed coupling reactions with various alkynes to give rise to the corresponding 3 alkynyl-5-bromo-2-pyrones with good to excellent chemical yields and regioselectivity. PMID- 11922811 TI - Controlled photocyclization, photodimerization, and photoisomerization of stilbazole salts within Nafion membranes. AB - [reaction: see text] Water- and methanol-swollen Nafion membranes were used as microreactors to successfully control the photochemical reaction pathway of stilbazole derivatives. Of particular interest is the production of azaphenanthrene (the product not obtainable in homogeneous solution) in high yield under high bulk concentration. PMID- 11922812 TI - Self-assembly and characterization of supramolecular [60]fullerene-containing 2,6 diacylamidopyridine with uracil derivative by hydrogen-bonding interaction. AB - [structure: see text] Novel self-assembly systems of uracil derivatives with organofullerene by a three-point hydrogen-bonding interaction were designed and established. The formation of hydrogen bonding was established by 1H NMR studies in CDCl3. PMID- 11922813 TI - Convergent synthesis of the E'FGH' ring fragment of ciguatoxin 1B via an acetylene cobalt complex strategy. AB - [reaction: see text] A convergent synthesis of the E'FGH' ring fragment of ciguatoxin has been accomplished through (i) coupling between the E' ring acetylide and the H' ring-aldehyde, (ii) stereoselective F ring cyclization via an acetylene cobalt complex, (iii) conversion to a carbonyl function, and (iv) reductive hydroxy-ketone cyclization to construct the G ring. PMID- 11922814 TI - Efficient loading of sulfonamide safety-catch linkers by Fmoc amino acid fluorides. AB - [reaction: see text] Fmoc-protected amino acid fluorides were found to be excellent reagents for the acylation of sulfonamide safety-catch linkers (SCL) suitable for the subsequent preparation of peptide C-terminal thioesters. High loadings were obtained on different types of resins with low levels of epimerization. PMID- 11922816 TI - Generation of the first tellurium-containing diheteropentalene. AB - [reaction: see text] The known diester dibromomethylthiophene was converted to the tellurolo[3,4-c]thiphene, the first example of a diheteropentalene containing a tellurophene annelated across the 3,4-c position to thiophene. It is stable in solution and adds to DMAD across the 2,5-positions in tellurophene. The intermediate loses tellurium and collapses to a tetracarbomethoxy benzo[c]thiophene derivative. PMID- 11922815 TI - Pyran annulation: asymmetric synthesis of 2,6-disubstituted-4-methylene tetrahydropyrans. AB - [reaction: see text] A reaction process for the asymmetric construction of a variety of cis or trans disubstituted pyrans is described. This sequences allows for the asymmetric convergent union of two aldehydes with silyl-stannane reagent 1 in a two-step process: catalytic asymmetric allylation of the first aldehyde using 1 with a BITIP catalyst, followed by reaction of the alcohol so obtained with a second aldehyde and TMSOTf. PMID- 11922817 TI - A new ligand scaffold for catalytic asymmetric alkylzinc additions to aldehydes. AB - [reaction: see text] 1,3-Azole derivatives of 2-aminocyclohexanecarboxylic acid represent a new class of bidentate ligands for metal-mediated catalytic asymmetric synthesis. N-[2-(4-Isopropyl-4,5-dihydrooxazol-2 yl)cyclohexyl]methanesulfonamide (6) is particularly well suited for the addition of alkylzinc reagents to aliphatic aldehydes in high enantiomeric excess. PMID- 11922818 TI - Preparation of substituted piperazinones via tandem reductive amination-(N,N' acyl transfer)-cyclization. AB - [reaction: see text] A one-pot, tandem reductive amination-transamidation cyclization reaction was employed to produce substituted piperazin-2-ones in good yields. Various amino acid methyl esters and transferable acyl groups were examined to establish the reaction's scope. PMID- 11922819 TI - Reversing the stereoselectivity of the intermolecular Pauson-Khand reaction: formation of endo-fused norbornadiene adducts. AB - [reaction: see text] An unprecedented endo-selective and regioselective intermolecular Pauson-Khand reaction takes place when heterobimetallic (Mo-Co) complexes derived from N-(2-alkynoyl)oxazolidinones or sultams are heated in the presence of norbornadiene. PMID- 11922820 TI - Glutathionyl transferase catalyzed addition of glutathione to COMC: a new hypothesis for antitumor activity. AB - [reaction: see text] Data are presented indicating that the potent antitumor activity of 2-crotonyloxymethyl-(4R,5R,6R)-4,5,6-trihydroxy-2-cyclohexenone (COTC) and 2-crotonyloxymethyl-2-cyclohexenone (COMC) is not likely the result of glyoxalase I inhibition, as has long been assumed. An alternative hypothesis is presented, based on the finding that COMC is a substrate for human glutathionyl transferase, which produces a transient, highly electrophilic glutathionylated 2 exomethylenecyclohexanone that can covalently modify proteins and nucleic acids. PMID- 11922821 TI - The N-hydroxymethyl group for stereoselective conjugate addition: application to the synthesis of (-)-statine. AB - [reaction: see text] Efficient synthesis of enantiomerically pure (-)-statine was achieved with the stereoselective intramolecular conjugate addition of the hydroxyl group tethered to the amino group of a configurationally stable N-Boc-L leucinal derivative. PMID- 11922822 TI - A novel photorearrangement of a cyclohexadiene derivative of C60. AB - [reaction: see text] Photorearrangement of tetraalkoxycarbonyl-substituted cyclohexadiene derivatives of C(60) yields not only well-known bis(fulleroid) but also bis(methano)fullerene. Existence of a labile and structurally new intermediate is observed in the reaction mixture. The discovery of the compound suggests the existence of another possible pathway giving those two products other than the widely accepted [4 + 4]/[2 + 2 + 2] mechanism. PMID- 11922823 TI - Catalytic, asymmetric trans-selective hetero Diels-Alder reactions using a chiral zirconium complex. AB - [reaction: see text] The first catalytic, asymmetric 2,3-trans-selective hetero Diels-Alder reaction has been developed. The reactions of aldehydes with Danishefsky's dienes proceeded smoothly to afford the pyranone derivatives in high yields with high trans-selectivities and enantioselectivities in the presence of a chiral zirconium complex, which was prepared from zirconium tert butoxide and (R)-3,3'-diiodobinaphthol or its derivatives, primary alcohol, and a small amount of water. This reaction was applied to the concise synthesis of (+) prelactone C. PMID- 11922824 TI - Synthesis of 2-C-methyl-D-erythritol 2,4-cyclopyrophosphate. AB - [reaction: see text] The synthesis of 2-C-methyl-D-erythritol 2,4 cyclopyrophosphate, a biochemical intermediate in the deoxyxylulose pathway of isoprenoid biosynthesis, was accomplished in four steps. Bisphosphorylation of 2 C-methyl-D-erythritol 1,3-diacetate, followed by carbodiimide cyclization and deprotection, led to the title compound in 42% overall yield. PMID- 11922825 TI - Highly diastereoselective nitrone cycloaddition onto a chiral ketene equivalent: asymmetric synthesis of cispentacin. AB - [reaction: see text] A highly diastereoselective intramolecular nitrone cycloaddition onto a chiral ketene equivalent, obtained by Horner-Wadsworth Emmons olefination of either enantiomer of bis-sulfinyl phosphonate 6, is described. Cycloaddition gave 5,5-disubstituted isoxazolidine 10 in good yield as a single diastereomer. Catalytic hydrogenolysis of 10 furnished either enantiomer of optically pure cis-2-aminocyclopentane-1-carboxylic acid. PMID- 11922826 TI - Fast synthesis of aryl triflates with controlled microwave heating. AB - [reaction: see text] Synthesis of aryl triflates from phenols using N phenyltriflimide requires only 6 min for completion when conducted with controlled microwave heating. The methodology was applied to both solution- and solid-phase conditions. Ten different aryl triflates were synthesized and isolated in good yields. Applications in high-throughput chemistry are suggested. PMID- 11922827 TI - N-Carbamate-assisted stereoselective synthesis of chiral vicinal amino sulfides. AB - [reaction: see text] Simply mixing amino alcohol A and thiol in toluene and TFA provided the corresponding amino sulfide B in excellent chemical yield and diastereoselectivity. A double SN2 process initiated by O-5 participation of the neighboring N-carbamate group was advanced to explain the overall retention of configuration at the chiral benzylic center. PMID- 11922828 TI - Lewis acid promoted phenylseleno group transfer tandem radical cyclization reactions. AB - [reaction: see text] A new Lewis acid promoted phenylseleno group transfer tandem radical cyclization method was developed. In the presence of Lewis acids such as Yb(OTf)3 or Mg(ClO4)2, under photolysis condition at low temperature (-45 degrees C), various unsaturated alpha-phenylseleno beta-keto amides underwent radical cyclization reactions to give monocyclic or bicyclic products in a highly efficient, regioselective, and stereoselective manner. PMID- 11922832 TI - Protein folding: is hierarchical versus nonhierarchical a productive issue? AB - A coarse-grained simulation accessing relevant folding timescales for beta lactoglobulin was corroborated experimentally and reveals a dynamic role for nonnative structures dictated by local propensity vis-a-vis the large-scale context. This picture prompts us to shift focus, leaving aside the hierarchical vs. nonhierarchical controversy. PMID- 11922833 TI - Pathway diversity and concertedness in protein folding: an ab-initio approach. AB - Making use of an ab-initio folding simulator, we generate in vitro pathways leading to the native fold in moderate size single- domain proteins. The assessment of pathway diversity is not biased by any a priori information on the native fold. We focus on two study cases, hyperthermophile variant of protein G domain (1gb4) and ubiquitin (1ubi), with the same topology but different context dependence in their native folds. We demonstrate that a quenching of structural fluctuations is achieved once the proteins find a stationary plateau maximizing the number of highly protected hydrogen bonds. This enables us to identify the folding nucleus and show that folding does not become expeditious until a concerted event takes place generating a topology able to prevent water attack on a maximal number of hydrogen bonds. This result is consistent with the standard nucleation mechanism postulated for two-state folders. Pathway diversity is correlated with the extent of conflict between local structural propensity and large-scale context, rather than with contact order: In highly context-dependent proteins, the success of folding cannot rely on a single fortuitous event in which local propensity is overruled by large-scale effects. We predict mutational Pi values on individual pathways, compute ensemble averages and predict extent of surface burial and percentage of hydrogen bonding on each component of the transition state ensemble, thus deconvoluting individual folding-route contributions to the averaged two-state kinetic picture. Our predicted kinetic isotopic effects find experimental support and lead to further probes. Finally, the molecular redesign potentiality of the method, aimed at increasing folding expediency, is explored. PMID- 11922834 TI - Homology modeling of falcipain-2: validation, de novo ligand design and synthesis of novel inhibitors. AB - Increasing resistance of malaria parasites, in particular Plasmodium falciparum, demands a serious search for novel targets. Cysteine protease in P. falciparum, encoded by a previously unidentified gene falcipain 2, provides one such target to design chemotherapeutic agents for treatment of malaria. In fact, a few cysteine protease inhibitors have been shown to inhibit growth of cultured malarial parasites. In absence of a crystal structure for this enzyme, homology modeling proved to be a reasonable alternative to study binding requirements of the enzyme. A homology model for falcipain 2 was developed and validated by docking of known vinyl sulfone inhibitors. Further, based on the observations of these studies, novel isoquinoline inhibitors were designed and synthesized, which exhibited in vitro enzyme inhibition at micromolar concentrations. PMID- 11922835 TI - Molecular dynamics simulations of conformational behavior of linear RGD peptidomimetics and cyclic prodrugs in aqueous and octane solutions. AB - Conformations available to a class of cyclic prodrugs and corresponding linear RGD peptidomimetics were explored using 1 ns length molecular dynamics simulations performed with the program CHARMM. Water and octane, modeled explicitly, were used as solvents to mimic the change of the environment experienced by the solutes upon partition from water to membrane in the trans cellular transport process. In water, the linear peptidomimetics tended to populate extended-like structures, characterized by strong favorable interactions with solvent and low intrinsic stability. In these extended conformations the charged termini are able to assume large distances, above 15 A for the longest systems. These linear peptidomimetics have been found to exhibit the highest potency in experimental studies, in accord with the trends experimentally observed for RGD peptides. In contrast, in octane compact conformers of the linear peptidomimetics were favored, with all charged groups aggregated and shielded from solvent, exhibiting high intrinsic stability and weak solute solvent interactions. Our calculations predict a large unfavorable energy change for transferring the linear systems from water to octane, in agreement with experimental findings that these compounds are not transported via the trans cellular pathway. The cyclic prodrugs did not exhibit major structural differences between the simulations in water and octane, adopting turn-like conformations in both solvents. The limited response of the cyclic structures to changes in the environment leads to energies of transfer from water to octane that are also unfavorable, but much less so than for the linear molecules. This effect is in accord with the observed enhanced passive trans-cellular transport of the cyclic prodrugs. PMID- 11922836 TI - Structural and ICAM-1-docking properties of a cyclic peptide from the I-domain of LFA-1: an inhibitor of ICAM-1/LFA- 1-mediated T-cell adhesion. AB - The purpose of this work was to study the conformation of cyclic peptide 1, cyclo(1,12)-Pen1-Ile2-Thr3-Asp4-Gly5-Glu6-Ala7- Thr8-Asp9-Ser10-Gly11-Cys12-OH, derived from the I-domain of the LFA-1 alpha-subunit. We found that cyclic peptide 1 can bind to the D1-domain of ICAM-1 and inhibit ICAM-1/LFA-1-mediated homotypic and heterotypic T-cell adhesion. To understand the bioactive conformation and binding requirements for cyclic peptide 1, its solution structure was studied using NMR, CD, and molecular dynamics simulations. Furthermore, possible binding properties between the cyclic peptide and the D1 domain of ICAM-1 were evaluated using docking experiments. This cyclic peptide has a stable betaII -turn at Asp4- Gly5-Glu6-Ala7 and a betaI-turn at Pen1-Ile2 Thr3-Asp4; a less stable betaV-turn is found at the C-terminal region. The beta turn at Asp4- Gly5-Glu6-Ala7 was also found in the X-ray structure of the I domain of LFA-1. Our CD studies showed that the peptide binds to calcium/magnesium and forms a 1:1 (peptide:calcium/magnesium) complex with low cation concentrations and multiple types of complexes with higher cation concentrations. Binding to divalent cations causes a conformational change in peptide 1; this is consistent with our previous study that binding of peptide 1 to ICAM-1 was influenced by divalent cations. Docking studies show the interaction between cyclic peptide 1 and the D1-domain of ICAM-1; it indicates that the Ile2-Thr3-Asp4-Gly4-Glu6-Ala7-Thr8 sequence interacts with the F and C strands of the D1-domain. Finally, these studies will help us design a new generation of selective peptides that may bind better to the D1-domain of ICAM-1. PMID- 11922837 TI - Molecular dynamics simulations of calcium-free calmodulin in solution. AB - A 4-ns molecular dynamics simulation of calcium-free calmodulin in solution has been performed, using Ewald summation to treat electrostatic interactions. Our simulation results were mostly consistent with solution experimental studies, including NMR, fluorescence and x-ray scattering. The secondary structures within the N- and C-terminal domains were conserved in the simulation, with trajectory structures similar to the NMR-derived model structure 1CFD. However, the relative orientations of the domains, for which there are no NMR restraints, differed in details between the simulation and the 1CFD model. The most interesting information provided by the simulations is that the dynamics of calcium-free calmodulin in solution is dominated by slow rigid body reorientations of the domains. The interdomain distance fluctuated between 29 and 39 A, and interdomain orientation angle, defined as the pseudo-dihedral formed by the four calcium binding sites, varied between -2 degrees and 108 degrees. Similarly, the domain linker region also exhibited significant fluctuations, with its length varying in the 34-45 A range and its bend angle in the 10-100 degrees range. The simulations are in accord with fluorescence results suggesting that calcium-free calmodulin is more compact and more flexible than the calcium activated form. Surprisingly, quite similar solvent accessibilities of the hydrophobic patches were seen in the calcium-free trajectory described in this work and previously generated calcium loaded calmodulin simulations. Thus, our simulations suggest a reexamination of the standard model of the structural change of calmodulin upon calcium binding, involving exposure of the hydrophobic patches to solvent. PMID- 11922838 TI - Molecular dynamics study of Ca(2+) binding loop variants of silver hake parvalbumin with aspartic acid at the "gateway" position. AB - The helix-loop-helix (i.e., EF-hand) Ca(2+) ion binding motif is characteristic of a large family of high-affinity calcium ion binding proteins, including the parvalbumins, oncomodulins and calmodulins. In this work we describe a set of molecular dynamics computations on the major parvalbumin from the silver hake (SHPV-B) and on functional fragments of this protein, consisting of the first four helical regions (the ABCD fragment), and the internal helix-loop- helix region (the CD fragment). In both whole protein and protein fragments (i.e., ABCD and CD fragments), the 9th loop residue in the calcium ion binding site in the CD helix-loop-helix region (the so-called "gateway" position) has been mutated from glutamic acid to aspartic acid. Aspartic acid is one of the most common residues found at the gateway position in other (non-parvalbumin) EF- hand proteins, but has never been found at the gateway position of any parvalbumin. (Interestingly, aspartic acid does occur at the gateway position in the closely related rat and human oncomodulins.) Consistent with experimental observations, the results of our molecular dynamics simulations show that incorporation of aspartic acid at the gateway position is very disruptive to the structural integrity of the calcium ion coordination site in the whole protein. The aspartic acid mutation is somewhat less disruptive to the calcium ion coordination sites in the two parvalbumin fragments (i.e., the ABCD and CD fragments), presumably due to the higher degree of motional freedom allowable in these protein fragments. One problem associated with the E59D whole protein variant is a prohibitively close approach of the aspartate carboxyl group to the CD calcium ion observed in the energy-minimized (pre-molecular dynamics) structure. This steric situation does not emerge during energy-minimization of the wild-type protein. The damage to the structural integrity of the calcium ion coordination site in the whole protein E59D variant is not relieved during the molecular dynamics simulation. In fact, during the course of the 300 picosecond simulation, all of the calcium ion ligands leave the primary coordination sphere. In addition, the conserved hydrogen- bonds (in the short beta-sheet structure) that links the CD site to the symmetry-related EF site (in the non-mutated whole protein) is also somewhat disrupted in the E59D whole protein variant. These results suggest that the Ca(2+) ion binding deficiencies in the CD loop are related, at least in part, to the unique interaction that exists between the paired CD and EF hands in the whole protein. Our theoretical results correlate well with previous studies on engineered EF-hand proteins and with all of our experimental evidence on whole silver hake parvalbumin and enzymatically-generated parvalbumin fragments. PMID- 11922839 TI - Molecular dynamics simulation of 7, 8-dihydro-8-oxoguanine DNA. AB - To elucidate the effect of guanine lesion produced by the oxidative damage on DNA, 1 nanosecond molecular dynamics simulations of native and oxidized DNA were performed. The target DNA molecules are dodecamer duplex d(CGCGAATTCGCG)(2) and its derivative duplex d(C(1)G(2)C(3)(8 oxoG)(4)A(5)A(6)T(7)T(8)C(9)G(10)C(11)G(12).d(C(13)G(14)C(15)G(16)A(17)A(18)T(19) (20)C(21)G(22)C(23)G(24), which has one oxidized guanine, 7, 8-dihydro-8 oxoguanine (8-oxoG), at the fourth position. The local structural change due to the lesion of 8-oxoG and the global dynamic structure of the 8-oxoG DNA were studied. It was found that the 8-oxoG DNA remained structurally stable during the simulation due to newly produced hydrogen bonds around the (8-oxoG)(4) residue. However, there were distinguishable differences in structural parameters and dynamic property in the 8-oxoG DNA. The conformation around the (8-oxoG)(4) residue departed from the usual conformation of native DNA and took an unique conformation of epsilon-zeta in B(II) conformation and chi in high anti orientation at the (8-oxoG)(4) residue, and adopted a very low helical twist angle at the C(3):G(22)-(8-oxoG)(4):C21) step. Further analysis by principal component analysis indicated that the formation of the hydrogen bonds around the (8-oxoG)(4) residue plays a role as a trigger for the conformational transition of the 8-oxoG DNA in the conformational space. PMID- 11922840 TI - The structure and dynamics of the U4/U6 snRNP: implications for pre-mRNA splicing and use as a model system to investigate the RNA-mediated effects of (5F)Ura. AB - Pre-mRNA splicing is one of the most complex and intricate processes in eukaryotic cell biology. Over the past decade, my laboratory has been interested in determining the structures of RNA components of the spliceosome, and in investigating how the structure, stability and dynamics of these RNA components are perturbed by nucleoside analog substitution. In particular, we have investigated the U4/U6 snRNA complex as a model system for understanding the biophysical basis for the RNA-mediated effects of the widely-used anticancer drug 5-fluorouracil ((5F)Ura). In this review, our studies that have provided novel information concerning the structure of U4 snRNA and its interactions with U6 snRNA and the Sm (or common) snRNA binding proteins are summarized. These studies have also quantified the structural and thermodynamic consequences of (5F)Ura in this model system. Our work to date provides the foundation on which future studies investigating the biophysical basis for spliceosomal assembly and for clarifying the mechanisms of anticancer drugs targeted at nucleic acid-mediated processes will be developed. PMID- 11922841 TI - Explicit solvent molecular dynamics simulation of duplex formed by the modified oligonucleotide with alternating phosphate/phosphonate internucleoside linkages and its natural counterpart. AB - Impact of the internucleoside linkage modification by inserting a methylene group on the ability of the modified oligonucleotide to hybridize with a natural DNA strand was studied by fully solvated molecular dynamics (MD) simulations. Three undecamer complexes were analyzed: natural dT(11).dA(11) duplex as a reference and two its analogs with alternating modified and natural linkages in the deoxyadenosine chain. The isopolar, non-isosteric modified linkages were of 5'-O PO(2)-CH(2)-O-3' (5'PC3') or 5'-O-CH(2)-PO(2)-O-3' (5'CP3') type. Simulations were performed by using the AMBER 5.0 software package with the force field completed by a set of parameters needed to model the modified segments. Both modifications were found to lead to double helical complexes, in which the thymidine strand as well as deoxyriboses and unmodified linkages in the adenosine strand adopted conformations typical for the B-type structure. For each of the two conformational richer modified linkages two stable conformations were found at 300 K: the -ggt and ggt for the 5'PC3' and ggg, tgg for the 5'CP3', respectively. Both modified chains adopted helical conformations with heightened values of the inclination parameter but without affecting the Watson-Crick hydrogen bonds. PMID- 11922842 TI - Mechanical model of the nucleosome and chromatin. AB - A theoretical framework for evaluating the approximate energy and dynamic properties associated with the folding of DNA into nucleosomes and chromatin is presented. Experimentally determined elastic constants of linear DNA and a simple fold geometry are assumed in order to derive elastic constants for extended and condensed chromatin. The model predicts the Young s modulus of extended and condensed chromatin to within an order of magnitude of experimentally determined values. Thus we demonstrate that the elastic properties of DNA are a primary determinant of the elastic properties of the higher order folded states. The derived elastic constants are used to predict the speed of propagation of small amplitude waves that excite an extension(sound), twist, bend or shear motion in each folded state. Taken together the results demonstrate that folding creates a hierarchy of time, length and energy scales. PMID- 11922843 TI - Poly(rA).poly(rU) with Ni(2+) ions at different temperatures: infrared absorption and vibrational circular dichroism spectroscopy. AB - Phase transitions were studied of the sodium salt of poly(rA).poly(rU) induced by elevated temperature without Ni(2+) and with Ni(2+) in 0.07 M concentration in D(2)O (approximately 0.4 [Ni]/[P]). The temperature was varied from 20 degrees C to 90 degrees C. The double-stranded conformation of poly(rA).poly(rU) was observed at room temperature (20 degrees C-23 degrees C) with and without Ni(2+) ions. In the absence of Ni(2+) ions, partial double- to triple-strand transition of poly(rA).poly(rU) occurred at 58 degrees C, whereas only single- stranded molecules existed at 70 degrees C. While poly(rU) did not display significant helical structure, poly(rA) still maintained some helicity at this temperature. Ni(2+) ions significantly stabilized the triple-helical structure. The temperature range of the stable triple-helix was between 45 degrees C and 70 degrees C with maximum stability around 53 degrees C. Triple- to single-stranded transition of poly(rA).poly(rU) occurred around 72 degrees C with loss of base stacking in single-stranded molecules. Stacked or aggregated structures of poly(rA) formed around 86 degrees C. Hysteresis took place in the presence of Ni(2+) during the reverse transition from the triple-stranded to the double stranded form upon cooling. Reverse Hoogsteen type of hydrogen-bonding of the third strand in the triplex was suggested to be the most probable model for the triple-helical structure. VCD spectroscopy demonstrated significant advantages over infrared absorption or the related electronic CD spectroscopy. PMID- 11922844 TI - Low temperature solution structures and base pair stacking of double helical d(CGTACG)(2). AB - Solution structures and base pair stacking of a self- complementary DNA hexamer d(CGTACG)(2) have been studied at 5, 10 and 15 degrees C, respectively. The stacking interactions among the center base pair steps of the DNA duplex are found to improve when the terminal base pairs became less stable due to end fraying. A new structural quantity, the stacking sum (Sigma(s)), is introduced to indicate small changes in the stacking overlaps between base pairs. The improvements in the stacking overlaps to maintain the double helical conformation are probably the cause for the observed temperature dependent structural changes in double helical DNA molecule. A detailed analysis of the helical parameters, backbone torsion angles, base orientations and sugar conformations of these structures has been performed. PMID- 11922845 TI - Flexible elements in the structure of promoter DNA as probed by cationic surfactant binding. AB - A susceptibility of promoter DNA for adaptive conformational transitions has been studied using a cationic surfactant dodecyltrimethylammonium bromide (C(12)TAB) as a model DNA-binding ligand. DNAse 1 and KMnO(4) were utilized as structure specific reagents. Both reagents revealed ligand-induced perturbations in the double helix of promoters T7A1 and T7D. These conformational transitions appeared to be strongly associated with pyrimidine-purine steps, which have non-random distribution within RNA polymerase contact region of the promoter DNA and are present in the binding sites for a majority of transcription regulation proteins. Potential flexibility of these elements creates therefore a specific media for transcription complex formation. Molecular mechanism of DNA interaction with C(12)TAB is discussed. PMID- 11922846 TI - Distinguishing "looped-out" and "stacked-in" DNA bulge conformation using fluorescent 2-aminopurine replacing a purine base. AB - The conformation of a bulged DNA base, whether looped-out of the DNA helix or stacked-in between the flanking bases, can be distinguished using fluorescence spectroscopy of an inserted fluorescent base. If 2-aminopurine, a structural analog of adenine and guanine, is placed in duplex DNA as the bulged base replacing an adenine or guanine, it loops out of the DNA helix into solution. This is determined by the decrease or increase of 2-aminopurine fluorescence during DNA thermomelting: if the 2-aminopurine base stacks into the helix, its fluorescence increases or remains about the same during DNA duplex melting, but if the 2-aminopurine base loops out of the helix, its fluorescence decreases upon melting of the DNA duplex. PMID- 11922847 TI - DNA binding properties of novel dansylated distamycin analogues in which the fluorophore is directly conjugated to the N-methyl-pyrrole. AB - Polyamides that are structural analogues of the naturally occurring DNA minor groove binding antibiotic distamycin (Dst) are promising candidates as gene modulators. Developing strategies for the large scale screening and monitoring of the cellular distribution of such ligands would aid the faster discovery of molecules, which would have eventual utility in molecular biology and medicine. Attachment of fluorescent tags would be a useful step towards this end. A fundamental question in this connection is whether the tag modifies the DNA binding affinity of the parent compounds. Towards answering this question, we have developed two oligopeptides that bear the dansyl (N, N dimethylaminonaphthalene sulfonamido fluorophore) coupled directly to the N terminus of the conjugated N-methylpyrrole carboxamide network, and possess three or four N-methyl pyrrole carboxamide units (abbreviated as Dn3 and Dn4 respectively). DNA binding abilities of these molecules were assessed from fluorescence titration experiments, duplex-DNA T(m) analysis (employing both UV and fluorescence spectroscopy), induced circular dichroism measurements (ICD), salt dependence of ICD and apparent binding constant measurements (K(app)) employing ethidium bromide (EtBr) displacement assay. Both these molecules reported DNA binding in the form of an enhanced fluorescence emission. As judged from the ICD measurements, salt dependence of ICD, T(m) analysis and K(app) measurements, the binding affinities of the molecules that possessed dansyl group at their N-termini were lower than the ones with equivalent number of amide units, but possessed N-methylpyrrole carboxamide unit at their N- termini. These results would have implications in the future design of fluorescent polyamides. PMID- 11922849 TI - Identifying early decline of physical function in community-dwelling older women: performance-based and self-report measures. AB - BACKGROUND AND PURPOSE: The ability to identify early decline in physical function is important, but older people experiencing decline may fail to report the early changes in physical function. The purpose of this study was to compare the descriptions of physical function in community-dwelling older women obtained using performance-based and self-report measures. SUBJECTS AND METHODS: One hundred seventy community-dwelling women with a mean age of 74.3 years (SD=4.3, range=56.6-83.6) completed the activities of daily living (ADL), instrumental activities of daily living (IADL) and social activity (SA) sections of the Functional Status Questionnaire (FSQ). They also completed performance-based measures of gait speed and the 7-item Physical Performance Test (PPT). RESULTS: The majority of the women scored at the ceiling for the self-report measures of function (ADL=77%, IADL=61%, SA=94%), whereas only 7% scored at the ceiling for the PPT and 30% scored at the ceiling for gait speed (defined as >1.2 m/s). For 2 items of the FSQ, sensitivity was low (8% and 9%) and specificity was high (97% and 98%) compared with performance on the PPT. DISCUSSION AND CONCLUSION: In this sample of community-dwelling older women, performance-based measures identified more limitations in physical function than did self-report measures. PMID- 11922850 TI - The development and testing of APTA Clinical Performance Instruments. American Physical Therapy Association. AB - BACKGROUND AND PURPOSE: The purposes of this article are to describe the process of developing the physical therapist (PT) and physical therapist assistant (PTA) Clinical Performance Instruments (CPIs) and to present the available information on the psychometric properties of each instrument. SUBJECTS: Two hundred seventeen PTA students and 282 PT students participated in the pilot studies of the CPIs, and 181 PTA students and 319 PT students participated in field studies. METHODS: To construct each instrument, content was first gathered from a variety of instruments and American Physical Therapy Association documents related to PT and PTA practice and education. Data compiled during the pilot and field study phases of the project led to the construction of the fourth (final) versions of the CPIs, which although not studied are currently in use. RESULTS: Intraclass correlation coefficients (ICC [2,1]) measuring the interrater reliability of the CPI total score were good (ICC=.87) for the PT total score and moderate (ICC=.77) for the PTA total score. Construct validity was supported by the substantial differences in mean CPI score for students completing first as compared with final clinical experiences, by the correlation between CPI item scores and total days of clinical experience, and by the lack of correlation with the Social Skills Inventory score. DISCUSSION AND CONCLUSION: Sale of the fourth (final) versions of the PT CPI occurred in November 1997 and of the PTA CPI in March 1998. Data based on psychometric evaluation of the final version have not yet been collected and reported. In the task force's opinion, the third drafts can provide reliable and valid measurements of PT or PTA student clinical performance. The fourth versions were based on this iteration. PMID- 11922851 TI - The effect of transcutaneous electrical stimulation on spinal motor neuron excitability in people without known neuromuscular diseases: the roles of stimulus intensity and location. AB - BACKGROUND AND PURPOSE: The Hoffmann reflex (H-reflex) is widely acknowledged as an indirect indicator of spinal motor neuron excitability. The purpose of this study was to determine whether transcutaneous electrical stimulation (TES), applied over the dorsiflexors or plantar flexors of the ankle, would alter the soleus muscle's H-reflex. Attention was focused on the roles of stimulus intensity and location. SUBJECTS: Thirty-two volunteers without known neuromuscular diseases (17 women [53%]; mean years of age=27, SD=7.3, range=21 48) were studied. METHODS: Subjects were randomly assigned to 1 of 4 groups, and TES was administered for 15 minutes. Stimulation site and intensity varied according to group assignment. H-reflexes were recorded before and for 10 minutes after TES. RESULTS: H-reflex amplitudes increased following TES at sensory threshold, whereas H-reflex amplitudes did not change following TES at 1.5 times motor threshold. The site of stimulation did not influence the resulting H reflexes. DISCUSSION AND CONCLUSION: Low-intensity TES increases H-reflex amplitudes (and presumably the excitability of spinal motor neurons to Ia afferent input) in subjects without known neuromuscular diseases. High-intensity TES had little influence on H-reflex amplitudes. PMID- 11922852 TI - Reliability of safe maximum lifting determinations of a functional capacity evaluation. AB - BACKGROUND AND PURPOSE: Functional capacity evaluations (FCEs) are measurement tools used in predicting readiness to return to work following injury. The interrater and test-retest reliability of determinations of maximal safe lifting during kinesiophysical FCEs were examined in a sample of people who were off work and receiving workers' compensation. SUBJECTS: Twenty-eight subjects with low back pain who had plateaued with treatment were enrolled. Five occupational therapists, trained and experienced in kinesiophysical methods, conducted testing. METHODS: A repeated-measures design was used, with raters testing subjects simultaneously, yet independently. Subjects were rated on 2 occasions, separated by 2 to 4 days. Analyses included intraclass correlation coefficients (ICCs) and 95% confidence intervals. RESULTS: The ICC values for interrater reliability ranged from.95 to.98. Test-retest values ranged from.78 to.94. DISCUSSION AND CONCLUSION: Inconsistencies in subjects' performance across sessions were the greatest source of FCE measurement variability. Overall, however, test-retest reliability was good and interrater reliability was excellent. PMID- 11922853 TI - Agility and perturbation training for a physically active individual with knee osteoarthritis. AB - BACKGROUND AND PURPOSE: People with knee osteoarthritis (OA) sometimes report episodes of knee instability that limit their activities of daily living. The episodes of instability are similar to those reported by people with knee ligament injuries. The authors believe that modifications of interventions that are used to promote knee stability in individuals with knee ligament injuries can be used to enhance knee stability and function in people with knee OA. The purpose of this case report is to describe the development and implementation of an agility and perturbation training program that was used in conjunction with traditional rehabilitation activities for a patient with knee OA. CASE DESCRIPTION: The patient was a 73-year-old woman with a diagnosis of bilateral knee OA. Her chief complaints were knee pain and episodes of partial "giving way" at the knee during walking, stair climbing, and participation in tennis and golf. The patient participated in 12 treatment sessions at a frequency of 2 visits per week. The rehabilitation program consisted of lower-extremity stretching, strengthening and endurance exercises that were supplemented with a variety of walking-based agility training techniques, and perturbation training techniques. OUTCOME: At the completion of rehabilitation, the patient was able to walk on level surfaces and stairs and return to playing golf and tennis without episodes of instability and with reduced pain. DISCUSSION: Supplementing rehabilitation programs for people with knee OA with a modified agility and perturbation training program may assist them in returning to higher levels of physical activity with less pain and instability following rehabilitation. Further research is needed to determine the role of agility and perturbation training in people with knee OA. PMID- 11922854 TI - Tissue adaptation to physical stress: a proposed "Physical Stress Theory" to guide physical therapist practice, education, and research. AB - The purpose of this perspective is to present a general theory--the Physical Stress Theory (PST). The basic premise of the PST is that changes in the relative level of physical stress cause a predictable adaptive response in all biological tissue. Specific thresholds define the upper and lower stress levels for each characteristic tissue response. Qualitatively, the 5 tissue responses to physical stress are decreased stress tolerance (eg, atrophy), maintenance, increased stress tolerance (eg, hypertrophy), injury, and death. Fundamental principles of tissue adaptation to physical stress are described that, in the authors' opinion, can be used to help guide physical therapy practice, education, and research. The description of fundamental principles is followed by a review of selected literature describing adaptation to physical stress for each of the 4 main organ systems described in the Guide to Physical Therapist Practice (ie, cardiovascular/pulmonary, integumentary, musculoskeletal, neuromuscular). Limitations and implications of the PST for practice, research, and education are presented. PMID- 11922855 TI - Therapeutic potential of serotonin antagonists in depressive disorders. AB - Although the precise neurochemical imbalances in depression are still unknown, a role for the neurotransmitter 5-hydroxytryptamine (serotonin) has been implicated since the identification of the first effective antidepressants, imipramine and iproniazid. This led to the development of the selective serotonin re-uptake inhibitors which are widely used in the treatment of depression and depressive disorders, including generalised anxiety disorder, social phobia, obsessive compulsive disorder etc. Studies involving chronic administration in rats led to the hypothesis that the different classes of antidepressant treatment produce a common neuroadaptive change, namely an enhancement of serotonin neurotransmission, albeit via different pre and postsynaptic mechanisms. From this, it was suggested that serotonin antagonists should induce similar neuroadaptive changes, either directly or through a potentiation of other antidepressant agents. Here, the profiles of novel serotonin antagonists currently in preclinical development are reviewed and their therapeutic potential is assessed. PMID- 11922856 TI - Non-surgical management of spinal cord injury. AB - Spinal cord injury remains a devastating neurological condition with limited therapeutic opportunities. Since decompressive surgery and high-dose methylprednisolone have limited utility for most patients, spinal cord injury clearly represents a major medical challenge. Experimental evidence has suggested that secondary cellular injury processes may be a realistic target for therapeutic intervention with the goal of inhibiting the progression of detrimental changes that normally follows traumatic injury to the cord. Preventing or reducing this delayed cellular injury may alone improve neurological recovery or facilitate future regenerative approaches to the injured cord. This review summarises recent advances in the development of pharmacological agents targeting the acute phase of spinal cord injury as well as potential strategies to facilitate regeneration of the spinal cord. PMID- 11922857 TI - Emerging strategies for drug development in motor neuron disease. AB - Until recently, attempts to identify disease-modifying treatments in motor neuron disease have largely failed. This was due to a lack of understanding of the disease pathogenesis and poor clinical trial design. Recent advances in the genetics of motor neuron disease and the maturation of tissue culture techniques have increased our understanding of the mechanisms of cell death in motor neuron disease. This has in turn led to better in vitro and transgenic animal models, allowing for a more systematic study of the disease process and analysis of potential therapeutic agents. With the advent of these models, motor neuron disease drug research has entered a new era. It is now possible to pursue mass screening of compounds in the in vitro models. Successful compounds in vitro can be advanced to the transgenic animal models. Once toxicity and potential therapeutic effects in animals are known, agents which remain promising can be taken to Phase I, II and III clinical trials. This will increase the likelihood of success and is more desirable than methods previously utilised. A hierarchical approach such as this will be even more important as multi-drug regimens are to be studied. The number of combinations for these multi-drug regimens becomes prohibitive without a screening method. PMID- 11922858 TI - The potential of dopamine agonists in drug addiction. AB - The use of dopamine agonists in alcohol, stimulant and nicotine dependence has been examined. The direct agonists, such as bromocriptine and pergolide, have not shown utility in alcohol or cocaine abuse and dependence in larger controlled trials. Indirect agents, such as selegiline, may be helpful in cocaine or nicotine abuse and larger clinical trials are underway. Disulfiram may also raise dopamine levels and has shown promise for cocaine dependence. Other indirect agents, such as mazindol and methylphenidate, have not proven effective for cocaine addiction but have not been tested in alcohol or nicotine abuse. Agents for subtypes of dopamine receptors, such as D3, and the use of partial agonists may be useful future treatment approaches. Animal studies also suggest that tailoring treatment to subgroups of patients based on genotype may improve responses. PMID- 11922859 TI - Treating restless legs syndrome: current pathophysiological concepts and clinical trials. AB - Restless legs syndrome is a distinctive clinical syndrome with a prevalence of about 5% in the general population. One of the outstanding characteristics of restless legs syndrome is its extreme responsiveness to dopaminergic agents. Together with the latest pathophysiological and genetic findings, recent epidemiological and clinical data give a new insight into the classification of restless legs syndrome, thus building the theoretical foundation for the development of new pharmacological methods in its treatment. Current efforts within this area focus on establishing dopaminergic substances for therapy. The hypothesis of a disturbed iron metabolism in restless legs syndrome has been revived by recent theoretical considerations. The present review attempts to explain current strategies of treatment for restless legs syndrome in relation to aetiological, genetic and pathophysiological findings. PMID- 11922860 TI - Developments in the treatment of chronic hepatitis C. AB - Hepatitis C virus is the most common chronic, blood-bourne infection, affecting 170 million people worldwide, approximately 3% of the global population. Of those infected with hepatitis C virus, 50 - 85% will develop chronic hepatitis C. Although hepatitis C is primarily a disease of the liver, a diagnosis is currently defined by the presence of the hepatitis C virus and treatment success is defined by the clearance of the virus. IFN-alpha is currently the mainstay of chronic hepatitis C therapy; the antiviral and anti-inflammatory components of IFN target both the infectious and the hepatic manifestations of the disease. However, even in combination with ribavirin, interferon therapy is not fully efficacious. Recently, the search for a more effective treatment has led investigators to optimise interferon therapy by developing pegylated interferons. Challenges facing our current treatment of hepatitis C virus include lack of efficacy in patients with difficult-to-treat disease, such as patients with cirrhosis or infected with hepatitis C virus genotype 1 (who represent a majority of US hepatitis C virus infections), the toxicity of combination therapy, the expense and difficulty of therapy and the poor reception of these treatments by many patients. The development of new hepatitis C antiviral agents is critical to our management of this disease. A number of approaches are under investigation, including long-acting interferons, immunomodulators, antifibrotics, specific hepatitis C virus-derived enzyme inhibitors, drugs that either block hepatitis C virus antigen production from RNA or prevent normal processing of hepatitis C virus proteins and other molecular approaches to treating hepatitis C virus, such as ribozymes and antisense oligonucleotides. PMID- 11922861 TI - Recent developments in carbapenems. AB - Carbapenems are beta-lactam antibiotics characterised by the presence of a beta lactam ring with a carbon instead of sulfone in the 4-position of the thyazolidinic moiety. The first carbapenem to be utilised in therapy was imipenem, the N-formimidoyl derivative of thienamycin. Imipenem is coadministered with cilastatin, an inhibitor of human renal dehydropeptidase I, as imipenem is hydrolysed by this enzyme. Meropenem was the first carbapenem with a 1-beta methyl group and 2-thio pyrrolidinyl moiety, which renders this antibiotic stable to renal dehydropeptidase I. Other carbapenems for parenteral administration later discovered include biapenem, panipenem, ertapenem, lenapenem, E-1010, S 4661 and BMS-181139. Carbapenems which are orally administered include sanfetrinem, DZ-2640, CS-834 and GV-129606. Carbapenems have an ultra-broad spectrum of antibacterial activity and stability to almost all clinically relevant beta-lactamases. This differentiates them from all other currently available classes of beta-lactam antibiotics. However, Class B beta-lactamases, along with some rare Class A and D enzymes, are able to hydrolyse these antibiotics. Although Class B enzymes are generally chromosomally-encoded (isolated from Stenotrophomonas maltophilia, Aeromonas spp., Bacillus cereus, Bacteroides fragilis, Flavobacterium spp. and Legionella gormanii), plasmid metallo-beta-lactamases now are appearing in B. fragilis, Pseudomonas aeruginosa, Acinetobacter baumannii and members of Enterobacteriaceae such as Serratia marcescens and Klebsiella pneumoniae. The number of these enzymes compared to the number of other beta-lactamase types is still low, however, it is likely that they will spread due to the increased selective pressure of carbapenem use. The very broad spectrum of antimicrobial activity associated with a good clinical efficacy and a favourable safety profile makes the carbapenems valuable as 'first line' antibiotics in initial empirical therapy for the treatment of severe infections. PMID- 11922862 TI - Therapies in development for community-acquired pneumonia. AB - The current use of antimicrobials has become more complex due to the extensive emergence of antibiotic resistance. The single most important approach in resistance control is probably the judicious use of chemotherapeutic agents. New agents that may be of use in the treatment of community-acquired pneumonia are currently in development. Antimicrobials can be grouped according to their mechanism of action. These include protein synthesis inhibitors (ketolides, oxazolidinones, streptogramins and glycylcyclines), nucleic acid synthesis inhibitors (fluoroquinolones), peptidoglycan synthesis inhibitors (beta-lactams and glycopeptides) and agents interfering with membrane function (cationic peptides and lipopeptides). Among those agents under development, only the oxazolidinones, the cationic peptides and the lipopeptide antibiotics can be truly regarded as structurally novel inhibitors as the other agents are analogues of existing compounds which have been in clinical use for many years. PMID- 11922863 TI - Pharmaceutical thrombosis prevention in cardiovascular disease. AB - Cardiovascular diseases are a leading cause of morbidity and mortality in modern society. As a result of this, great efforts have been made to establish regimens for prophylaxis and treatment of such disorders. Pharmacological intervention is also a prerequisite for the success of other therapeutic approaches, e.g. coronary angioplasty. Prevention of platelet aggregation is a goal that can be achieved by counteracting various receptors on the platelet surface. The main attentions for such interventions are focused on inhibiting the glycoprotein IIb/IIIa receptor. So far, they are limited to intravenous usage. Adenosine diphosphate receptor inhibitors are available for intravenous and oral usage. Their effect is, at least partly, also exerted via the counteraction of adenosine diphosphate-mediated activation of the glycoprotein IIb/IIIa complex. An oral direct thrombin inhibitor is under clinical evaluation. This review focuses on atherothrombotic disorders, but recent advances within new fields of anticoagulation (i.e., treatment of severe septic shock and a novel approach to prevent thromboembolic disorder during surgery) should not be overlooked. PMID- 11922864 TI - Pimagedine: a novel therapy for diabetic nephropathy. AB - Hyperglycaemia is directly involved in causing long-term diabetic complications. The non-enzymatic glycation of proteins, yielding irreversible advanced glycation end products and advanced glycation end products-derived protein crosslinking, participates in the development of diabetic complications, such as diabetic nephropathy. Diabetic nephropathy is becoming a major medical problem with increasing numbers of these patients progressing to end stage renal disease, thus requiring renal replacement therapy. While several interventions may slow the development and progression of diabetic nephropathy, there is no effective treatment to prevent or reverse the disease. Pimagedine (aminoguanidine HCl) has been shown to be an effective agent in reducing the severity of the structural and functional alterations associated with experimental diabetic nephropathy. Preliminary studies suggest a beneficial effect of pimagedine in treating patients with diabetic nephropathy. In summary, these observations support a role for advanced glycation end products inhibitors, like pimagedine, in the management of diabetic nephropathy, either alone or in combination with other therapies. PMID- 11922865 TI - Nerve growth factor selectively regulates expression of transcripts encoding ribosomal proteins. AB - BACKGROUND: NGF exerts a variety of actions including promotion of neuronal differentiation and survival. The PC12 rat pheochromocytoma cell line has proved valuable for studying how NGF works and has revealed that the NGF mechanism includes regulation of gene expression. Accordingly, we used SAGE (Serial Analysis of Gene Expression) to compare levels of specific transcripts in PC12 cells before and after long-term NGF exposure. Of the approximately 22,000 transcripts detected and quantified, 4% are NGF-regulated by 6-fold or more. Here, we used database information to identify transcripts in our SAGE libraries that encode ribosomal proteins and have compared the effect of NGF on their relative levels of expression. RESULTS: Among the transcripts detected in our SAGE analysis, 74 were identified as encoding ribosomal proteins. Ribosomal protein transcripts were among the most abundantly expressed and, for naive and NGF-treated PC12 cells, represented 5.2% and 3.5%, respectively, of total transcripts analyzed. Surprisingly, nearly half of ribosomal protein transcripts underwent statistically significant NGF-promoted alterations in relative abundance, with changes of up to 5-fold. Of the changes, approximately 2/3 represented decreases. A time course revealed that the relative abundance of transcripts encoding RPL9 increases within 1 hr of NGF treatment and is maximally elevated by 8 hr. CONCLUSIONS: These data establish that NGF selectively changes expression of ribosomal protein transcripts. These findings raise potential roles for regulation of ribosomal protein transcripts in NGF-promoted withdrawal from the cell cycle and neuronal differentiation and indicate that regulation of individual ribosomal protein transcripts is cell- and stimulus-specific. PMID- 11922867 TI - Elevated antibody to D-alanyl lipoteichoic acid indicates caries experience associated with fluoride and gingival health. AB - : BACKGROUND: Acidogenic, acid-tolerant bacteria induce dental caries and require D-alanyl glycerol lipoteichoic acid (D-alanyl LTA) on their cell surface. Because fluoride inhibits acid-mediated enamel demineralization, an elevated antibody response to D-alanyl LTA may indicate subjects with more acidogenic bacteria and, therefore, an association of DMFT with fluoride exposure and gingival health not apparent in low responders. METHODS: Cluster analysis was used to identify low antibody content. Within low and high responders (control and test subjects), the number of teeth that were decayed missing and filled (DMFT), or decayed only (DT) were regressed against fluoride exposure in the water supply and from dentrifice use. The latter was determined from gingival health: prevalences of plaque (PL) and bleeding on probing (BOP), and mean pocket depth (PD). Age was measured as a possible confounding cofactor. RESULTS: In 35 high responders, DMFT associated with length of exposure to fluoridated water (F score), PL and BOP (R2 = 0.51, p < 0.001), whereas in 67 low D-ala-IgG responders, DMFT associated with PL, age, and PD (R2 = 0.26, p < 0.001). BOP correlated strongly with number of 7 7 decayed teeth (DT) in 54 high responders (R2 = 0.57, p < 0.001), but poorly in 97 low responders (R2 = 0.12, p < 0.001). The strength of the PD association with DMFT, or of BOP with DT, in high responders significantly differed from that in low responders (p < 0.05). CONCLUSION: Caries associates with gingival health and fluoridated water exposure in high D-alanyl LTA antibody responders. PMID- 11922866 TI - Respiratory syncytial virus and TNF alpha induction of chemokine gene expression involves differential activation of Rel A and NF-kappa B1. AB - BACKGROUND: Respiratory syncytial virus (RSV) infection of airway epithelial cells stimulates the expression and secretion of a variety of cytokines including the chemotactic cytokines interleukin-8 (IL-8), monocyte chemoattractant protein 1 (MCP-1), and RANTES (regulated upon activation, normal T cell expressed and secreted). Chemokines are important chemoattractants for the recruitment of distinct sets of leukocytes to airway sites of inflammation. RESULTS: We have shown previously that chemokine expression is regulated in airway epithelial cells (A549) in a stimulus-specific manner in part through the redox-responsive transcription factors AP-1 and NF-kappaB. In this study, we examined the NF kappaB-mediated effects of RSV and the proinflammatory cytokine TNFalpha on the induction of IL-8, MCP-1 and RANTES chemokine gene expression in A549 epithelial cells. The results demonstrate that RSV induces chemokine expression with distinct kinetics that is associated with a specific pattern of NF-kappaB binding activity. This distinction was further demonstrated by the differential effects of the NF-kappaB inhibitors dexamethasone (DEX) and N-acetyl-L-cysteine (NAC). NAC preferentially inhibited RSV induced chemokine expression, whereas DEX preferentially inhibited TNFalpha induced chemokine expression. DNA binding studies using NF-kappaB subunit specific binding ELISA demonstrated that RSV and TNFalpha induced different NF-kappaB binding complexes containing Rel A (p65) and NF-kappaB1 (p50). Both TNFalpha and RSV strongly induced Rel A the activation subunit of NF-kappaB, whereas only TNFalpha was able to substantially induce the p50 subunit. Consistent with the expression studies, RSV but not TNFalpha induction of Rel A and p50 were markedly inhibited by NAC, providing a mechanism by which TNFalpha and RSV can differentially activate chemokine gene expression via NF-kappaB. CONCLUSIONS: These data suggest that RSV induction of chemokine gene expression, in contrast to TNFalpha, involves redox-sensitive NF-kappaB complexes containing predominantly Rel A. PMID- 11922868 TI - A nomenclature for X-linked amelogenesis imperfecta. AB - Mutations of the X-chromosome amelogenin gene (AMELX) are associated with amelogenesis imperfecta (AI) phenotypes (OMIM no. 301200). Currently, 12 different AMELX mutations have been identified in individuals with abnormal enamel characteristic of AI. A notable feature of AI is the variable clinical phenotype, spurring interest in genotype-phenotype correlations. It is important that researchers and clinicians have an informative and reliable means of reporting and communicating these molecular defects. Therefore, the purpose here was to present a systematic nosology for reporting the genomic, cDNA and protein consequences of AMELX mutations associated with AI. The proposed nomenclature adheres to conventions proposed for other conditions and can be adopted for the autosomal forms of AI as the molecular basis of these conditions becomes known. PMID- 11922870 TI - A postero-anterior videofluorographic study of the intra-oral management of food in man. AB - Recent studies of the manipulation and reduction of food in the human mouth have used lateral-projection videofluorography (VFG) and so have concerned antero posterior food movement. To determine the degree to which (a) food was chewed bilaterally as opposed to unilaterally, (b) the frequency of side changes, (c) the contribution of tongue and cheek activity to food manipulation, and (d) the effect of food consistency on these activities, nine young adults were recorded eating 7g cubes of four foods (banana, biscuit [cookie] and two types of meat: "tender" and "tough") in the postero-anterior projection. Videotapes recorded at 30 frames/s were acquired to disk; data were analysed as single frames and in slow-motion. As expected, the meat samples, being fibrous, required more chewing and manipulation, allowing details of the process to be established. Food was ingested in the midline; the tongue then positioned the cube on the occlusal plane of one side by a combination of pushing, tilting and twisting movements before any occlusal contact (stage I transport). During processing, food was kept on the occlusal surface by a combination of rhythmic tongue-pushing moving the food buccally (41% of cycles), and cheek-pushing (28% of cycles) returning it in the lingual direction. This reciprocating movement ensured that different parts of the food were subjected to occlusal force in successive cycles. Bilateral chewing was common and associated with either: (a) a near-symmetrical closing movement with tooth-food-tooth contact occurring almost simultaneously on both sides, or (b) an identifiable "active side" but a jaw movement extended medially to carry the lower molars through a power stroke on the "balancing" side. During the sequence, food requiring further chewing might be moved to the erstwhile balancing-side (balancing-side shift); or across the mouth to the tooth row or vestibule on the other side and "stored" for later reduction (segregation shift). Towards the end of the sequence, triturated material was moved to the midline (aggregation shift) for bolus formation and deglutition. While distinct patterns of mediolateral and vertical jaw movements seem to be associated with shift and transport cycles, these connections have not yet been established with sufficient robustness to support predictions of intra-oral events from jaw movement profiles alone. PMID- 11922869 TI - Amelogenesis imperfecta phenotype-genotype correlations with two amelogenin gene mutations. AB - Amelogenin, the predominant matrix protein in developing dental enamel, is considered essential for normal enamel formation, but its exact functions are undefined. Mutations in the AMELX gene that encodes for amelogenin protein cause X-linked amelogenesis imperfecta (AI), with phenotypes characterized by hypoplastic and/or poorly mineralized enamel. Eight different AMELX deletion and substitution mutations have been reported to date. The purpose here was to evaluate the genotype and phenotype of two large kindreds segregating for X linked AI. Phenotypically affected males in family 1 had yellowish-brown, poorly mineralized enamel; those in family 2 had thin, smooth, hypoplastic enamel. Heterozygous females in both kindreds had vertical hypoplastic grooves in their enamel. DNA was obtained from family members; exons 1-7 of AMELX were amplified and sequenced. Mutational analysis of family 1 revealed a single-base-pair change of A-->T at nucleotide 256, resulting in a His-->Leu change. Analysis of family 2 revealed deletion of a C-nucleotide in codon 119 causing a frameshift alteration of the next six codons, and a premature stop codon resulting in truncation of the protein 18 amino acids shorter than the wild-type. To date, all mutations that alter the C-terminus of amelogenin after the 157th amino acid have resulted in a hypoplastic phenotype. In contrast, other AMELX mutations appear to cause predominantly mineralization defects (e.g. the mutation seen in family 1). This difference suggests that the C-terminus of the normal amelogenin protein is important for controlling enamel thickness. PMID- 11922871 TI - Nanoindentation mapping of the mechanical properties of human molar tooth enamel. AB - The mechanical behavior of dental enamel has been the subject of many investigations. Initial studies assumed that it was a more or less homogeneous material with uniform mechanical properties. Now it is generally recognized that the mechanical response of enamel depends upon location, chemical composition, and prism orientation. This study used nanoindentation to map out the properties of dental enamel over the axial cross-section of a maxillary second molar (M(2)). Local variations in mechanical characteristics were correlated with changes in chemical content and microstructure across the entire depth and span of a sample. Microprobe techniques were used to examine changes in chemical composition and scanning electron microscopy was used to examine the microstructure. The range of hardness (H) and Young's modulus (E) observed over an individual tooth was found to be far greater than previously reported. At the enamel surface H>6GPa and E>115GPa, while at the enamel-dentine junction H<3GPa and E<70GPa. These variations corresponded to the changes in chemistry, microstructure, and prism alignment but showed the strongest correlations with changes in the average chemistry of enamel. For example, the concentrations of the constituents of hydroxyapatite (P(2)O(5) and CaO) were highest at the hard occlusal surface and decreased on moving toward the softer enamel-dentine junction. Na(2)O and MgO showed the opposite trend. The mechanical properties of the enamel were also found to differ from the lingual to the buccal side of the molar. At the occlusal surface the enamel was harder and stiffer on the lingual side than on the buccal side. The interior enamel, however, was softer and more compliant on the lingual than on the buccal side, a variation that also correlated with differences in average chemistry and might be related to differences in function. PMID- 11922872 TI - Movement of temporomandibular joint tissues during mastication and passive manipulation in miniature pigs. AB - Movement is an important aspect of the biomechanics of the temporomandibular joint (TMJ). To track the relative movements of TMJ components, radio-opaque markers were implanted in the left squamosal bone, mandible and retrodiscal tissue of miniature pigs. Medial-lateral (ML) and dorsal-ventral (DV) fluoroscopic records were made 8-10 weeks later during chewing and passive manipulation. Marker movements were digitized from the videotapes. During passive manipulation, the deformation of the lateral capsule was also measured with a differential variable-reluctance transducer. The results provide new details about porcine chewing pattern, which is distinguished by a regularly alternating chewing side. During masticatory opening, the mandible had a centre of rotation (CR) well inferior to the condyle and close to the angle. In contrast, the passive opening movement showed a higher CR location close to the condylar neck, indicating a different motion from masticatory opening. The retrodiscal tissue followed the movements of the mandibular condyle during both mastication and passive manipulation. The lateral capsule elongated during ipsilateral shifts and retrusion, implying a possible role in limiting such movements. These movement characteristics provide a useful reference for studies on the TMJ using pigs. PMID- 11922873 TI - Mercury distribution and concentration in rats fed powdered dental amalgam. AB - The aim was to evaluate the distribution and elimination of mercury in the tissues of rats exposed to powdered dental amalgam. The study comprised of three groups: the first (A) was an unexposed control, the second (B) was fed 8.3mg of powdered amalgam weekly for 12 weeks, and the third (C) was fed 25.0mg of powdered amalgam weekly for 4 weeks followed by either a 4- or 8-week period of non-exposure. The total amount of amalgam administered in each of the latter two groups was 0.1g. Animals were killed at 4, 8 and 12 weeks, and their kidneys, liver and brains were removed. Mercury present in tissues from one side was analysed by cold-vapour atomic absorption spectrometry; tissues from the other side were stained autometallographically. No mercury was detected in any tissue of control group A. In group B, the mercury concentration in the kidneys was significantly higher than that of the control, whereas in group C, the mercury concentration in both the liver and kidneys decreased significantly during the period of non-exposure. Mercury was not detected in brain tissue from any group. The light-microscopic findings were in agreement with the quantitative results. Mercury grains were most common in the renal cortex, especially in the proximal tubule. Although there was some mercury accumulation in the tissues of the two experimental groups, no pathological reactions were noted. These data suggest that, in the rat, the ingestion of dental amalgam in small quantities for a limited period does not lead to morphological changes in the liver, brain or kidneys. It seems likely that mercury does not accumulate in brain tissue and that any accumulation in the kidneys and liver is at least partially reduced after ingestion ceases. PMID- 11922874 TI - Bone geometry in cercopithecoid mandibles. AB - This study explores the relation between cortical bone geometry in the mandibular corpus and in vivo masticatory stress patterns and dietary specialization in cercopithecoid primates. Cortical bone distribution in the mandibles of three species of Old World monkeys (Macaca fascicularis, Procolobus badius, Lophocebus albigena) was measured by computed tomography. The arrangement of bone within sections was quantified as (1) the ratio of cortical area to the enclosed (total) subperiosteal area; (2) the ratio of orthogonal second moments of area; and (3) size-adjusted measures of cortical area and regional thickness. Cross-sectional geometry differed among samples, but consistent patterns of cortical thinning and bone area were found within individual sections. This consistency was despite the marked differences in diet and feeding behavior that distinguish the three taxa. Lingually thin and basally thick cortical bone was found in the three monkeys; previously published data suggest that this pattern may be stereotypical among anthropoid primates. It is hypothesized that the interactive effects of shear, bending and torsion produce eccentric loads in corpus sections, which are mirrored by this asymmetrical arrangement of cortical bone. When interpreted against existing data for other primate groups, these results are consistent with the hypothesis that masticatory-loading profiles are broadly similar across anthropoids despite the distinctive occlusions found among the suborder. Understanding of the impact of diet on jaw morphology is, therefore, not improved by considerations of cortical bone distribution, i.e. the inference of diet from jaw form is best predicated on considerations of relative corpus size rather than cross-sectional geometry. PMID- 11922875 TI - Identification of mouse submaxillary gland protein in mouse saliva and its binding to mouse oral bacteria. AB - The mouse submaxillary gland protein (mSMGP) is highly expressed in the submandibular gland of the adult mouse and rat. It shares 51% identity at the amino-acid level with a human protein, the prolactin-inducible protein (PIP)/gross cystic disease fluid protein 15 (GCDFP-15), which has been found in saliva, tears, sweat, seminal plasma, submucosal glands of the lung and amniotic fluid. More recently, the human PIP has been reported to bind to bacterial strains normally found in the mouth, ear canal and human skin. Sequence analysis of mSMGP/PIP earlier identified the presence of a signal peptide, suggesting that it is a secreted protein. Here, by Western blotting, mSMGP/PIP has been identified in mouse saliva. To investigate further the role of this secreted protein, its ability to bind specifically to oral bacteria was examined; the hypothesis was that mSMGP/PIP is involved in non-immune host defence by binding to bacteria. Several bacterial strains, found to belong to the genera Streptococcus, Aerococcus, Pseudomonas, Staphylococcus, Sphingomonas, Vibrio and Aeromonas, were isolated from the mouse oral cavity. Following incubation of these bacteria with (35)S-labeled, in vitro-translated mSMGP/PIP, the protein was found to bind specifically and selectively to several but not all strains tested, showing the highest affinity for the streptococci. The protein also bound specifically to an Aerococcus sp., and a low binding interaction with the Pseudomonas and Staphylococcus spp. was observed. The conservation of SMGP sequences among several animal species suggests that this protein may play an important part in the biology of the submandibular gland. As the function of the mSMGP/PIP is still undetermined, these findings provide insight into a possible involvement of this protein in host defence. PMID- 11922876 TI - The effect of pulpward pressure on the response to 50% lidocaine (lignocaine) applied to exposed dentine in cats. AB - The responses of intradental nerves can be blocked by 50% lidocaine applied to exposed, etched dentine. The aim here was to analyse electrophysiologically the contribution of pulpward pressure to the anaesthetic effect of lidocaine applied to exposed dentine. In 17 adult cats with intact lower canines, approximately 1mm of the coronal tip of the canine was removed and the exposed dentine acid-etched. Pulpward pressure was applied to the exposed dentine through a chamber fixed to the tip of the tooth. Cervical and coronal electrodes of silver wire and Ag/AgCl mixture were placed, the cervical 2mm from the gingival margin of the canine and the coronal into the coronal pulp chamber. The inferior alveolar nerve was surgically exposed and single pulpal nerve fibres responding to electrical stimulation of the canine were identified. The chamber was filled with 50% lidocaine solution, and 0 (atmospheric pressure), 1.3, 5.0 or 10.0 kPa pressure was applied to the chamber for 20 min. Orthodromic and antidromic action potentials were compared before and after the lidocaine application. When lidocaine was applied at 0, 1.3 or 5.0 kPa (n=5 each) for 20 min, all the orthodromic responses to cathodal electrical stimulation with the coronal electrode were blocked. Almost all orthodromic responses (12/15) remained unblocked when the cathodal electrical stimulation was applied with the cervical electrode. Applying 10.0 kPa pressure to the lidocaine solution blocked all the pulpal nerve responses (n=5) to both stimulation modes. The amplitude of the antidromic compound action potential was significantly decreased when the applied pressure was increased. These results suggest that increased pulpward pressure may effectively force anaesthetic solution more rapidly through the dentinal tubules into the pulp. PMID- 11922877 TI - A potential role for thalamocingulate circuitry in human maternal behavior. AB - BACKGROUND: Little is known about the regional brain basis of human maternal behavior. To understand this better, we have been examining brain activity in mothers listening to infant cries. METHODS: We measured functional Magnetic Resonance Imaging brain activity in healthy, breastfeeding first-time mothers with young infants while they listened to infant cries, white noise control sounds, and a rest condition. Based on the thalamocingulate theory of maternal behavior and pilot work, we hypothesized that the cingulate, medial thalamus, medial prefrontal cortex, and right orbitofrontal cortex would display more activity with infant cries than with white noise (comparison 1) and would uniquely activate with the cries, meaning that these regions would display activity with cry minus rest but not with white noise minus rest (comparison 2). RESULTS: In hypothesized regions, the group displayed more activity in the medial thalamus, medial prefrontal and right orbitofrontal cortices with both comparisons. The anterior and posterior cingulate cortex displayed more activity only with comparison 1. In non-hypothesized brain regions, several other structures thought important in rodent maternal behavior displayed activity with both comparisons including the midbrain, hypothalamus, dorsal and ventral striatum, and vicinity of the lateral septal region. CONCLUSIONS: Our results partially support our hypotheses and are generally consistent with neuroanatomical studies of rodent maternal behavior. PMID- 11922878 TI - Temporal coherence in ultradian sleep EEG rhythms in a never-depressed, high-risk cohort of female adolescents. AB - BACKGROUND: Previous work has indicated that low temporal coherence of ultradian sleep electroencephalographic rhythms is characteristic of depressed patients and of depressed women, in particular. It may also be evident in one quarter of those at high risk, based on a family history of depression. METHODS: The present study evaluated temporal coherence of sleep electroencephalographic rhythms in 41 adolescent girls with a maternal history of depression (high risk) and 40 healthy controls (low risk). The entire sample was followed clinically every 6 months for 2 years. RESULTS: Temporal coherence was significantly lower among the high-risk girls than in controls. Regression analyses predicted group from coherence values and correctly classified 70% of the high-risk group with a false-positive rate of 5% among controls. Moreover, 54% of the high-risk girls were identified with extreme low coherence. On clinical follow up, 14 girls showed depressive symptoms, 9 in the high-risk group (22.5%) and 5 controls (12.2%). Six met DSM-IV criteria for first-episode major depressive disorder, five high-risk and one control. Most importantly, 41% of those identified as having the most abnormal coherence values either showed symptoms of depression or met diagnostic criteria upon follow up. CONCLUSIONS: Low temporal coherence is evident in adolescent girls at high risk for depression. The more abnormal the coherence, the greater the risk of a first episode of major depressive disorder within 2 years of sleep study, approximately 10 times greater than in controls. PMID- 11922879 TI - The effects of an orally administered cholinergic agonist on REM sleep in major depression. AB - BACKGROUND: Centrally active cholinergic agents such as arecoline and physostigmine shorten rapid eye movement (REM) latency, reduce REM interval times, or both and do so preferentially in patients with depression. We tested an orally administered cholinergic agonist (donepezil HCL 10 mg [Aricept]) to determine whether this agent also alters REM timing in depressed patients (n = 8) compared with age- and gender-matched control subjects (n = 8). METHODS: All subjects were studied for 3 consecutive nights in the sleep laboratory. The design was a fixed-order placebo-donepezil protocol to accommodate the long half life of donepezil. Night 1 served as an adaptation night. On night 2, placebo was administered at 8:00 PM. On night 3, donepezil was administered at 8:00 PM. RESULTS: The cholinergic challenge distinguished the groups. In depressed patients REM latency was reduced compared with baseline (47.6 vs. 64.4, p =.04) following administration of donepezil. Control subjects showed no response: REM latency after donepezil was virtually identical to baseline REM latency (71.7 vs. 69.3). CONCLUSIONS: These data indicate that donepezil is likely to be useful in testing hypotheses related to cholinergic function in mood disorders. PMID- 11922880 TI - Plasma corticotropin-releasing factor in depressed patients before and after the dexamethasone suppression test. AB - BACKGROUND: Plasma cortisol, beta-endorphin, corticotropin, corticotropin releasing factor, and salivary cortisol concentrations, resting and after ingestion of 1 mg of dexamethasone, were investigated in depressed patients and controls. METHODS: Fourteen outpatients from the psychiatric department diagnosed with depressive disorder (ICD-10 Classification) participated in the study. The comparison group consisted of 12 healthy volunteers from the hospital staff. All hormones were measured using direct iodine-125 radioimmunoassay, except corticotropin-releasing factor, which included a sample preextraction and concentration step. RESULTS: The basal plasma cortisol and corticotropin releasing factor levels in depressive disorder were significantly higher than in the healthy group. After dexamethasone administration, corticotropin-releasing factor plasma values decreased significantly in the depressed group, but showed no significant changes in the controls. In depressive disorder baseline values correlated significantly for salivary cortisol and plasma cortisol, salivary cortisol and plasma corticotropin-releasing factor and plasma corticotropin and beta-endorphin. Similar correlations were found in the healthy subjects, except for salivary cortisol and plasma corticotropin-releasing factor. CONCLUSIONS: These findings indicate that the increased corticotropin-releasing factor plasma concentrations demonstrated in depressive disorder reflect the hypothalamic corticotropin-releasing factor hypersecretion evidenced in this illness. Therefore, measurements of plasma corticotropin-releasing factor levels can be considered a reliable tool for investigating the role of this peptide in the pathophysiology of depression. PMID- 11922881 TI - Monoamine depletion in unmedicated depressed subjects. AB - BACKGROUND: Although significant evidence suggests that diminished monoamine function is associated with clinical depression, catecholamine or indoleamine depletion alone has not been associated with significant mood changes in unmedicated depressed subjects or never-depressed control subjects. This study assesses the integrated role of these monoamine systems in depressed patients. METHODS: Unmedicated depressed subjects underwent a 2-week, double-blind, random ordered crossover study consisting of the following active and control conditions respectively: indoleamine (via tryptophan depletion) plus catecholamine (via alpha-methyl-paratyrosine administration) depletion and, separately, indoleamine plus sham (via diphenhydramine administration) catecholamine depletion. Ten subjects completed both conditions; two subjects were withdrawn after active testing and one after control testing. RESULTS: Mean Hamilton Depression Rating Scale (HDRS) scores decreased progressively throughout the study days (baseline 26.7 points +/- 1.7 SEM and termination 20.0 +/- 2.4, active depletion; baseline 26.1 points +/- 2.3 SEM and termination 23.2 +/- 2.6, control testing) but did not differ between groups. Only three patients demonstrated 20% or greater increases from baseline HDRS at any point during the observation days. CONCLUSIONS: Overall, results show that simultaneous disruptions of indoleamine and catecholamine function do not exacerbate symptoms in unmedicated depressed subjects, thus lending further support to the notion that monoamines regulate mood in actively depressed patients via indirect mechanisms. PMID- 11922882 TI - Effects of different frequencies of transcranial magnetic stimulation (TMS) on the forced swim test model of depression in rats. AB - BACKGROUND: Repetitive transcranial magnetic stimulation has been demonstrated in humans as well as in animal models to have an antidepressant effect, but the optimal frequency of stimulation is not known. We examined this question in a rat model of depression. METHODS: Young male Sprague-Dawley rats were allocated to two placebo (restraint and sham transcranial magnetic stimulation), one active control (imipramine), and four transcranial magnetic stimulation groups at 1, 5, 15 and 25 Hz and 1000 stimuli each. The Porsolt Swim Test was performed on day 1 (experiment 1). In an extension (experiment 2), the treatments were repeated on days 2 through 5, and the Swim Test repeated on days 3, 5, and 7. RESULTS: After one treatment session, all transcranial magnetic stimulation groups had significantly reduced immobility times compared with sham stimulation (p =.000), but the higher frequencies (15 and 25 Hz) did not differ significantly from lower (1 and 5 Hz) frequencies. After three sessions, all transcranial magnetic stimulation groups were different from placebo, and the rapid transcranial magnetic stimulation groups had lower immobility times than the slow transcranial magnetic stimulation groups (p =.035). After five sessions, only 15- and 25-Hz groups were different from control, and on day 7, only the 25-Hz group had reduced immobility. There was an overall difference between fast and slow transcranial magnetic stimulation (p =.010), and 1 Hz was different from the other three transcranial magnetic stimulation conditions (p =.016). CONCLUSIONS: Repetitive transcranial magnetic stimulation reduces immobility time in the Forced Swim Test model of depression, suggesting an antidepressant effect, which is evident at a range (1-25 Hz) of frequencies. With repeated administration, the findings suggest that the antidepressant effect of the higher frequencies, as for imipramine, is likely to be sustained, although the model used for this (i.e., repeating the Swim Test) requires further validation. PMID- 11922883 TI - Association between promoter polymorphic haplotypes of interleukin-10 gene and schizophrenia. AB - BACKGROUND: Schizophrenia is one of the most severe psychiatric disorders, with a worldwide incidence of 1%. Several reports show abnormal cytokine levels in psychotic patients and indicate a possible role of the immune response system in the pathogenesis of schizophrenia. Increased concentrations of interleukin 10 (IL 10) have been found in plasma of schizophrenic patients, suggesting its potential role as a candidate gene for susceptibility to schizophrenia. IL-10 gene maps on chromosome 1 (q31-q32), a locus associated with genetic susceptibility to schizophrenia. Three functional haplotypes of the gene (GCC, ACC, ATA) have been described, derived from different combinations of three "single nucleotide polymorphisms" and directly related to the expression levels of the protein. METHODS: We analyzed allele, genotype, and haplotype distributions in an association case-control study involving 106 schizophrenic patients and 143 unrelated healthy volunteers using polymerase chain reaction (PCR)-Single Strand Conformation Polymorphism and PCR Restriction Fragment Length Polymorphism methods. RESULTS: Our results show a significant increase of GCC homozygotes (the high IL-10-producing haplotype) in schizophrenic patients compared to control subjects (chi(2) = 13, p =.023; odds ratio = 3.03; 95% confidence interval, 1.274 7.355). CONCLUSIONS: These data could partly explain the abnormal secretion of IL 10 occurring in schizophrenic patients in response to infections or different stressors and suggest a potential role of IL-10 as a candidate gene for susceptibility to schizophrenia. PMID- 11922884 TI - Reduced communication between frontal and temporal lobes during talking in schizophrenia. AB - BACKGROUND: Communication between the frontal lobes, where speech and verbal thoughts are generated, and the temporal lobes, where they are perceived, may occur through the action of a corollary discharge. Its dysfunction may underlie failure to recognize inner speech as self-generated and account for auditory hallucinations in schizophrenia. METHODS: Electroencephalogram was recorded from 10 healthy adults and 12 patients with schizophrenia (DSM-IV) in two conditions: talking aloud and listening to their own played-back speech. Event-related electroencephalogram coherence to acoustic stimuli presented during both conditions was calculated between frontal and temporal pairs, for delta, theta, alpha, beta, and gamma frequency bands. RESULTS: Talking produced greater coherence than listening between frontal-temporal regions in all frequency bands; however, in the lower frequencies (delta and theta), there were significant interactions of group and condition. This finding revealed that patients failed to show an increase in coherence during talking, especially over the speech production and speech reception areas of the left hemisphere, and especially in patients prone to hallucinate. CONCLUSIONS: Reduced fronto-temporal functional connectivity may contribute to the misattribution of inner thoughts to external voices in schizophrenia. PMID- 11922886 TI - Spatial selective attention and inhibition in schizophrenia patients during acute psychosis and at 4-month follow-up. AB - BACKGROUND: Attentional abnormalities may lie at the core of cognitive symptoms in schizophrenia, but it is unclear how they relate to symptoms. The major aim of our study was to understand the relation between spatial attention and clinical symptoms from acute to chronic state. METHODS: Thirty-six acutely psychotic schizophrenia patients and 42 matched control subjects were assessed on three spatial attention measures: target location detection, interference (concurrent inhibition of distractor), and negative priming (subsequent inhibition of distractor). Symptoms were assessed by the Positive and Negative Syndrome Assessment Scale. Four months later, the same subjects were re-tested, and symptoms were re-assessed. RESULTS: Symptoms were significantly reduced at the follow-up. Schizophrenia patients were slower at detecting target location than control subjects, but they improved significantly over time. Schizophrenia patients and control subjects did not differ on the interference task. Negative priming was abolished during acute psychosis, but 4 months later it was restored. Positive symptoms were correlated with reduced negative priming but not with interference, nor with target detection. Negative priming during acute psychosis was significantly correlated with the clinical symptoms at the follow-up. CONCLUSIONS: These results suggest that reduced negative priming may be associated with increased clinical, symptoms especially the positive symptoms. PMID- 11922885 TI - Modulation of brain and serum glutamatergic concentrations following a switch from conventional neuroleptics to olanzapine. AB - BACKGROUND: Recent evidence suggests that effects upon glutamatergic transmission may contribute to the therapeutic action of certain atypical antipsychotic agents. METHODS: Glutamate concentrations were measured in serum and were estimated (Glx/Cr) in cingulate cortex by proton magnetic resonance spectroscopy (MRS) in schizophrenia patients while they were being treated with conventional neuroleptics and then 8 weeks after switching to olanzapine. Serum glutamate concentrations were obtained from 11 subjects, and MRS estimates of Glx/Cr were available from 10 subjects at both time points. RESULTS: Serum glutamate concentrations significantly increased after the switch from conventional agents to olanzapine; brain glutamate (Glx/Cr) did not change significantly; however, brain glutamate (Glx/Cr) concentrations increased significantly in patients who exhibited an improvement in negative symptoms with olanzapine compared with patients with no change or worsening of negative symptoms. CONCLUSIONS: Comparisons performed following the switch from conventional agents to olanzapine are consistent with previous studies of clozapine and provide additional preliminary evidence supporting the hypothesis that effects on excitatory amino acid activity may contribute to olanzapine's efficacy for treating negative symptoms. PMID- 11922887 TI - 3D laser surface scanning and geometric morphometric analysis of craniofacial shape as an index of cerebro-craniofacial morphogenesis: initial application to sexual dimorphism. AB - BACKGROUND: Over early fetal life, when disturbances in schizophrenia have been posited and craniofacial dysmorphogenesis reported, cerebral morphogenesis proceeds in embryological intimacy with craniofacial morphogenesis. Digitization technologies now allow 3D recording of craniofacial surface landmarks and modeling of craniofacial shape differences using geometric morphometrics. METHODS: Using normal sexual dimorphism as an exemplar, facial surfaces of 131 Medical School employees [82 females, 49 males] were recorded in 3D using a portable, hand-held laser scanner; 3D coordinate data were then analyzed using geometric morphometrics. RESULTS: Males and females differed markedly on an omnibus test of craniofacial shape. Logistic regression analysis of 16 principal components of shape variability, explaining 84.9% of the overall sample variance, generated 8 principal components as significant and independent discriminators. On visualization, the female face is wider and flatter; the eyes are more lateral, anterior and are further apart, and nasal bridge is posterior; the nose is smaller; the lips are fuller and the chin more forward. These findings are complementary to sexual dimorphism in cerebral structures. CONCLUSIONS: This technique reliably discriminates geometric features of craniofacial morphology that are associated with aspects of cerebral morphology, and may inform on putative neurodevelopmental disorders characterised by dysmorphogenesis. PMID- 11922888 TI - Johanson-Blizzard syndrome--a case study, behavioral manifestations, and successful treatment strategies. AB - Mental retardation has multiple causes, including genetic, and is often accompanied by severe behavioral difficulties. These behavioral problems frequently disrupt care, reduce quality of life, and represent risk of injury. In treating special populations where biologic interventions are most important, the psychiatrist must often combine appropriate pharmacologic interventions with behavioral strategies, employing staff support and using a multidisciplined team approach. This article reviews a case in which a person with Johanson-Blizzard syndrome, a rare genetic disorder whose physical manifestations have been described in the literature, is successfully treated using a combination of psychotropic medication and behavioral programming. Target behaviors of severe obsessive compulsive disorder and aggression were ameliorated using these strategies.Currently, specific pharmacologic interventions in combination with structured behavioral programming represents the most successful method of dealing with various forms of behavioral problems linked to mental retardation. PMID- 11922889 TI - Vitronectin is implicated as the matrix takes control of neointima formation. PMID- 11922890 TI - Altered Na/Ca exchange activity in cardiac hypertrophy and heart failure: a new target for therapy? AB - Increased Na/Ca exchange (NCX) expression may be part of the genetic reprogramming in cardiac remodeling. In this review we address the following questions: (1) Is increased NCX activity a general feature of cardiac remodeling in hypertrophy and heart failure? (2) How does this contribute to the contractile and electrical phenotype of hypertrophy and heart failure? (3) Should be consider NCX a potential therapeutic target? From a review of the literature we found that NCX activity can be increased, unchanged, or even downregulated during cardiac remodeling. When NCX activity is increased, it can be considered compensatory for contractile function, but with negative side-effects, including an increased risk of arrhythmias. Changes in activity do not necessarily reflect changes in gene expression. Altered NCX activity can also be a consequence of changes in other Ca(2+) fluxes or in [Na(+)](i) homeostasis. The role of NCX in contractile alterations and arrhythmogenesis varies with the different stimuli or stages of cardiac remodeling. Pharmacological block of NCX in heart failure or hypertrophy may thus be useful, but most likely only in specific conditions, perhaps as part of a combined approach. Development of drugs that target only a specific mode of the exchanger may offer a further advantage. PMID- 11922891 TI - Calcineurin and hypertrophic heart disease: novel insights and remaining questions. AB - In the past 2 years, an emerging body of research has focused on a novel transcriptional pathway involved in the cardiac hypertrophic response. Ever since its introduction, the significance of the calcineurin-NFAT module has been subject of controversy. The aim of this review is to provide both an update on the current status of knowledge and discuss the remaining issues regarding the involvement of calcineurin in hypertrophic heart disease. To this end, the molecular biology of calcineurin and its direct downstream transcriptional effector NFAT are discussed in the context of the genetic studies that established the existence of this signaling paradigm in the heart. The pharmacological mode-of-action and specificity of the calcineurin inhibitors cyclosporine A (CsA) and FK506 is discussed, as well as their inherent limitations to study the biology of calcineurin. A critical interpretation is given on studies aimed at analyzing the role of calcineurin in cardiac hypertrophy using systemic immunosuppression. To eliminate the controversy surrounding CsA/FK506 usage, recent studies employed genetic inhibitory strategies for calcineurin, which confirm the pivotal role for this signal transduction pathway in the ventricular hypertrophy response. Finally, unresolved issues concerning the role of calcineurin in cardiac pathobiology are discussed based upon the information available, including its controversial role in cardiomyocyte viability, the reciprocal relationship between myocyte Ca(2+) homeostasis and calcineurin activity and the relative importance of calcineurin in relation to other hypertrophic signaling cascades. PMID- 11922892 TI - Tumor necrosis factor-alpha and myocardial remodeling in progression of heart failure: a current perspective. AB - A milestone in the progression of congestive heart failure (CHF) is myocardial remodeling. Left ventricular (LV) remodeling during the progression of CHF is accompanied by changes in the structure of the myocardial extracellular matrix. Recent clinical and experimental studies have noted that increased release of tumor necrosis factor alpha (TNF-alpha) can contribute to LV myocardial remodeling. Experimental studies have noted that the induction of TNF-alpha can result in LV dilation and proceed to LV pump dysfunction. The biological effects of TNF-alpha are mediated through TNF receptors that are present on all nucleated cells in the heart. TNF receptor activation can induce a number of cellular and molecular events which contribute to LV remodeling in CHF, and include changes in myocyte size and viability and alterations in myocardial structure/composition. In vitro studies have demonstrated that TNF receptor activation can cause the induction of a proteolytic system. This proteolytic system, the matrix metalloproteinases (MMPs), is upregulated in models of LV dysfunction and possesses the capacity to degrade a wide variety of extracellular matrix components. Therefore, one pathway by which TNF-alpha can influence LV myocardial remodeling is through the induction of a specific portfolio of MMP species. Future basic and clinical studies which directly alter TNF receptor activity and measure myocardial MMP species and the relation to LV remodeling will provide new insight into this disease process and future therapeutic modalities. PMID- 11922893 TI - Relation between local myocardial growth and blood flow during chronic ventricular pacing. AB - Several studies have shown that, per unit mass, myocardial blood flow (MBF) and oxygen consumption are similar in hypertrophic and non-hypertrophic ventricles. This observation may be explained by the degree of myocardial growth matching the increase in oxygen demand. Such matching may, however, not be perfect at the local level, because substantial heterogeneity of MBF exists within the ventricular wall. We investigated to what extent local growth and MBF are matched after redistribution of workload within the left ventricular (LV) wall. Redistribution of workload was established by ventricular pacing at physiological heart rate, which induces asynchronous activation and contraction. Local wall mass (2D-echocardiography) and MBF (fluorescent microspheres) were determined in the canine LV wall before (t=0) and after 6 months of normal sinus rhythm (SHAM group, n=5) or 6 months of pacing at the LV free wall (PACE group, n=8). During acute pacing MBF (ml/min/g) increased with increasing distance to the pacing site. Local relative MBF (rMBF, local MBF normalized to mean MBF in the LV wall) varied from 0.8 adjacent to the pacing site to 1.2 in remote regions. After 6 months of pacing these regional differences had disappeared, probably due to changes in wall mass, which increased with increasing distance to the pacing site (by up to 39+/-13%). In SHAM animals rMBF at t=0 correlated well with rMBF 6 months later (r=0.71). In PACE animals, however, this correlation was poor (r=0.33), because rMBF increased in regions close to the pacing site with initial rMBF<1 and rMBF decreased in regions remote from the pacing site with initial rMBF>1. CONCLUSIONS: After redistribution of workload within the LV wall as induced by ventricular pacing, local load-regulated growth tends to equalize MBF distribution, but local adaptation of MBF also depends on initial MBF. PMID- 11922895 TI - Increased effects of C-type natriuretic peptide on cardiac ventricular contractility and relaxation in guanylyl cyclase A-deficient mice. AB - OBJECTIVE: The natriuretic peptides (NPs), atrial (ANP), B-type (BNP), and C-type (CNP) natriuretic peptides as well as their respective receptor-guanylyl cyclases (GC-A for ANP and BNP, and GC-B for CNP) are expressed in the heart. However, the local role of NPs in the regulation of cardiac contractility and the mutual interactions of NPs remain controversial. In the present study we evaluated the effects of ANP and CNP on cardiac function of wild-type (GC-A +/+) and GC-A deficient (GC-A -/-) mice. METHODS: The effects of NPs and their molecular mechanisms were assessed in the isolated perfused mouse working heart preparation. RESULTS: In GC-A +/+ hearts, CNP exerted a biphasic action: an immediate increase in inotropy and lusitropy, followed by a slowly developing negative inotropic effect. These effects were mimicked by the cGMP-analogue, 8 pCPT-cGMP. In contrast, ANP did not affect cardiac function. In GC-A -/- hearts, the immediate contractile responses to CNP and 8-pCPT-cGMP were significantly enhanced. CNP increased cardiac cGMP levels and stimulated phospholamban (PLB) phosphorylation; the effect on PLB, but not cGMP, was enhanced in GC-A -/- hearts. In addition, cardiac expression of cGMP-dependent protein kinase (cGK I) was significantly increased in GC-A -/- mice. CONCLUSION: CNP exerts a biphasic, initially positive inotropic and lusitropic, then negative inotropic effect in isolated working mouse hearts. A putative mechanism contributing to the immediate contractile responses is cGMP/cGK I-dependent phosphorylation of PLB and subsequent activation of the sarcoplasmic reticulum Ca(2+)-pump. ANP has no direct effects on cardiac contractility but chronic absence of its receptor, GC A, results in increased responsiveness to CNP. PMID- 11922894 TI - Pyruvate dehydrogenase and the regulation of glucose oxidation in hypertrophied rat hearts. AB - OBJECTIVE: Coupling of glucose oxidation to glycolysis is lower in hypertrophied than in non-hypertrophied hearts, contributing to the compromised mechanical performance of hypertrophied hearts. Here, we describe studies to test the hypothesis that low coupling of glucose oxidation to glycolysis in hypertrophied hearts is due to reduced activity and/or expression of the pyruvate dehydrogenase complex (PDC). METHODS: We examined the effects of dichloroacetate (DCA), an inhibitor of PDC kinase, and of alterations in exogenous palmitate supply on coupling of glucose oxidation to glycolysis in isolated working hypertrophied and control hearts from aortic-constricted and sham-operated male Sprague-Dawley rats. It was anticipated that the addition of DCA or the absence of palmitate would promote PDC activation and consequently normalize coupling between glycolysis and glucose oxidation in hypertrophied hearts if our hypothesis was correct. RESULTS: Addition of DCA or removal of palmitate improved coupling of glucose oxidation to glycolysis in control and hypertrophied hearts. However, coupling remained substantially lower in hypertrophied hearts. PDC activity in extracts of hypertrophied hearts was similar to or higher than in extracts of control hearts under all perfusion conditions. No differences were observed between hypertrophied and control hearts with respect to expression of PDC, PDC kinase, or PDC phosphatase. CONCLUSIONS: Low coupling of glucose oxidation to glycolysis in hypertrophied hearts is not due to a reduction in PDC activity or subunit expression indicating that other mechanism(s) are responsible. PMID- 11922896 TI - Regression of cardiomyocyte hypertrophy in SHR following chronic inhibition of the Na(+)/H(+) exchanger. AB - OBJECTIVE: Experiments were performed to examine the effect of chronic inhibition of the Na(+)/H(+) exchanger isoform-1 (NHE-1) on cardiac hypertrophy of spontaneously hypertensive rats (SHR). METHODS: SHR were orally treated during 1 month with two different doses (0.3 and 3.0 mg/kg/day) of the NHE-1 inhibitor, cariporide, or nifedipine (10.0 mg/kg/day). RESULTS: The two doses of cariporide did not differ in their effects after 1 month of treatment, since both induced a slight decrease in systolic blood pressure (SBP) of approximately 6 mmHg and regression of the heart weight to body weight ratio (mg/g) from 3.28+/-0.05 to 3.04+/-0.05 (0.3 mg) and 2.99+/-0.10 (3.0 mg, P<0.05). Nifedipine, given for the same period, produced similar reduction in the hypertrophy index (3.03+/-0.05), but with a much greater decrease in arterial pressure (35.6+/-7.4 mmHg). Chronic treatment with cariporide induced a complete regression of the augmented cross sectional area of left ventricular myocytes without significant changes in collagen content, serum procollagen 1 propeptide levels or myocardial distensibility. CONCLUSIONS: NHE inhibition represents a novel approach to induce regression of pathological hypertrophy of the heart. The finding can be rationalized mechanistically by previous in vitro studies suggesting a role of the NHE in the development of myocardial hypertrophy. PMID- 11922897 TI - Reduced swelling-activated Cl(-) current densities in hypertrophied ventricular myocytes of rabbits with heart failure. AB - OBJECTIVE: Hypertrophied myocytes of failing hearts have prolonged action potential durations. It is unknown how the swelling-activated Cl(-) current (I(Cl,swell)) affects the abnormal AP configuration. METHODS: We studied I(Cl,swell) in ventricular myocytes isolated from failing and age-matched normal rabbit hearts. We applied whole-cell patch-clamp methodology and activated I(Cl,swell) by lowering tonicity of the superfusate. RESULTS: Neither with ruptured-patch nor with amphotericin B perforated-patch, whole-cell clamp we found I(Cl,swell) active under isotonic conditions in either the normal or the hypertrophied failing heart (HFH) myocytes. I(Cl,swell) caused AP shortening and resting membrane potential (V(m)) depolarization in an osmotic gradient-dependent fashion. However, in the HFH myocytes swelling-induced AP changes were significantly smaller, even though the cells underwent the same relative change in planar cell surface area. Voltage-clamp experiments revealed that in HFH myocytes I(Cl,swell) current density was approximately 50% reduced. CONCLUSION: Reduced I(Cl,swell) densities in HFH myocytes cause limited AP shortening and V(m) depolarization upon swelling of the cells. PMID- 11922898 TI - Neuropeptide Y modifies the hypertrophic response of adult ventricular cardiomyocytes to norepinephrine. AB - OBJECTIVE: The hypertrophic response of adult rat cardiomyocytes to norepinephrine via alpha-adrenoceptor stimulation is limited by an inhibitory cross-talk of simultaneously beta-adrenoceptor stimulation. On the other hand, neuropeptide Y (NPY), known to be co-secreted with norepinephrine from intramural nerve endings of the heart, exerts an anti-beta-adrenergic effect. Therefore, it should be expected that NPY enhances the hypertrophic response to norepinephrine. This hypothesis was addressed in the present study. METHODS: Isolated adult ventricular cardiomyocytes from rats were used. As parameters of hypertrophic growth we investigated cell volume, cross-sectional area, protein mass. Protein and RNA synthesis were determined by incorporation of [(14)C]phenylalanine or [(14)C]uridine, respectively. RESULTS: Norepinephrine (1 micromol/l) did not significantly increase protein or RNA synthesis. In co-presence of NPY (100 nmol/l), however, norepinephrine increased protein synthesis by 44% and RNA synthesis by 18%. Under the same conditions, NPY enhanced the effect of norepinephrine on cell volume from +6.4 to +18.2%, its effect on cross-sectional area from +16 to +23%, and increased the protein/DNA ratio from 32.5 to 35.6 mg/mg. In parallel, norepinephrine caused a translocation of PKC-alpha and PKC delta into the particular fractions and this effect of norepinephrine was also enhanced by co-presence of NPY. In contrast, NPY did not enhance ERK-activation caused by norepinephrine. CONCLUSION: Our study indicates the anti-beta adrenergic effect of NPY is sufficient to modulate the hypertrophic response of adult ventricular cardiomyocytes to norepinephrine. The results suggest that the hypertrophic effect of norepinephrine via alpha-adrenoceptor stimulation can be modulated by co-release of NPY from intramural nerve endings. PMID- 11922899 TI - Role of P(1) purinergic receptors in myocardial ischemia sensory transduction. AB - OBJECTIVES: To characterize the role that cardiac sensory P(1) purinergic (adenosine A(1) or A(2)) receptors play in transducing myocardial ischemia. METHODS: Porcine nodose ganglion cardiac sensory neuron adenosine A(1) or A(2) receptor function was studied in situ during control states as well as in the presence of the peptides bradykinin and substance P or focal ventricular ischemia. The responses of porcine nodose ganglion cardiac and non-cardiac afferent neuronal somata to adenosine were also studied in vitro. RESULTS: Local application of A(1) or A(2) adenosine receptor agonists modified the activity generated by ventricular sensory neurites associated with 70 and 74% of identified nodose ganglion cardiac afferent somata in situ, respectively, exciting most neurons. In contrast, adenosine reduced the excitability of nodose ganglion cardiac afferent neuronal somata in vitro. Bradykinin and substance P affected 56 and 63%, respectively, of tested afferent neurons. The capacity of ventricular sensory neurites to transduce signals relating to these peptides was virtually eliminated by the presence of P(1) purinergic receptor antagonists. So was their capacity to transduce focal ventricular ischemia. Since most cardiac sensory neurites responded differently to adenosine in vivo than did cardiac afferent neuronal somata in vitro, it appears that the transduction properties of cardiac afferent neurons need to be characterized in situ. CONCLUSIONS: Most ventricular sensory neurites associated with nodose ganglion afferent neurons possess adenosine A(1) and/or A(2) receptors that play a primary role in transducing myocardial ischemic events to central neurons. These data support clinical observations implicating cardiac sensory purinoceptors in transducing myocardial ischemic events. PMID- 11922900 TI - Cardiotrophin-1 (CT-1) can protect the adult heart from injury when added both prior to ischaemia and at reperfusion. AB - OBJECTIVES: To determine whether the cytokine cardiotrophin-1 (CT-1) can protect the adult heart against ischaemia/reperfusion when added either prior to ischaemia or at reperfusion. BACKGROUND: CT-1 has previously been shown to protect cultured embryonic or neonatal cardiocytes from cell death. To assess the therapeutic potential of CT-1, it is necessary to determine whether this effect can be observed in adult cardiac cells both in culture and most importantly in the intact heart. METHODS: We examined the protective effect of CT-1 both in cultured adult rat cardiocytes and in the rat intact heart. In both cases, the cardiac cells were exposed to hypoxia/ischaemia followed by reoxygenation/reperfusion and CT-1 was administered either prior to hypoxia/ischaemia or at reoxygenation/reperfusion. RESULTS: CT-1 has a protective effect in reducing ischaemic damage in the intact heart ex vivo as assayed by infarct size to area at risk ratio (20% compared to 35%). Similar protective effects against cell death were noted in adult cells in vitro. Both in vitro and ex vivo CT-1 can exert a protective effect when added at the time of reoxygenation/reperfusion as well as prior to the hypoxic/ischaemic stimulus (cell death reduced from 50 to 20% in TUNEL assay, infarct size to zone at risk ratio reduced from 35 to 20%). These protective effects are blocked by an inhibitor of the p42/p44 MAPK pathway. CONCLUSION: CT-1 can protect adult cardiac cells both in vitro and in vivo when added both prior to or after the hypoxic/ischaemic stimulus. The potential therapeutic benefit of CT-1 when added at the time of reperfusion following ischaemic damage is discussed. PMID- 11922901 TI - Impact of HMG CoA reductase inhibition on small GTPases in the heart. AB - OBJECTIVE: Members of the Rho GTPase family, Rac1 and RhoA have been suggested to be mediators of cardiac hypertrophy in mice. Rho proteins are posttranslationally isoprenylated. In addition to cholesterol-lowering, statins inhibit the isoprenylation of small G proteins. Therefore, it was tested if these drugs inhibit Rac1 and RhoA activity in cardiomyocytes and, thereby, prevent angiotensin II-mediated expression of atrial natriuretic factor (ANF) and myosin light chain (MLC)-2 in the heart. METHODS AND RESULTS: Western and Northern analysis of rat neonatal cardiomyocytes and H9C2 cells showed inhibition of basal and angiotensin-stimulated Rac1 expression, membrane-translocation and activity after statin treatment. Similarly, basal and stimulated RhoA membrane expression was inhibited. Statins concentration- and time-dependently downregulated basal as well as angiotensin-induced expression of ANF by 86+/-2.3% and 89+/-1.7%, as well as MLC-2 by 75+/-4.1% and 84+/-6%, respectively. Direct inhibition of Rac GTPase by overexpression of the dominant negative mutant RacN17 or by Clostridium sordellii lethal toxin in rat H9C2 cells inhibited ANF expression by 70+/-4.9% and 78+/-10%, respectively. Inhibition of RhoA by Clostridium botulinum C3 transferase or the dominant negative mutant RhoN19 reduced ANF mRNA by 19+/-11% and 23+/-8%, respectively. To test these findings in vivo, spontaneously hypertensive rats were treated with atorvastatin, leading to a decrease in cardiac Rac1 and RhoA activity as determined by [35S]-GTP gamma S-binding assays by 61+/-16% and 72+/-24%, and downregulation of MLC-2 as well as ANF mRNA expression by 31+/-16% and 80+/-24%, respectively. CONCLUSIONS: (1) Statins downregulate the activity of small G proteins in cardiomyocytes in culture as well as in vivo. (2) Inhibition of Rac1 and RhoA by statins reduces myocardial expression of ANF and MLC-2. (3) Targeting myocardial Rho GTPases by statins may be a novel treatment strategy to prevent cardiac hypertrophy. PMID- 11922902 TI - Redistribution of connexin45 in gap junctions of connexin43-deficient hearts. AB - OBJECTIVE: Adult ventricular myocytes express two gap junction channel proteins: connexin43 (Cx43) and connexin45 (Cx45). Cx43-deficient mice exhibit slow ventricular epicardial conduction, suggesting that Cx43 plays an important role in intercellular coupling in the ventricle. Cx45 is much less abundant than Cx43 in working ventricular myocytes. Its role in ventricular conduction has not been defined, nor is it known whether expression or distribution of Cx45 is altered in Cx43-deficient mice. The present study was undertaken to determine (1) whether expression of Cx45 is upregulated and (2) whether gap junction structure and distribution are altered in Cx43-deficient mice. METHODS: Ventricular tissue from neonatal Cx43(+/+), Cx43(+/-) and Cx43(-/-) and adult Cx43(+/+) and Cx43(+/-) mice was analyzed by immunoblotting and confocal immunofluorescence microscopy. RESULTS: Total Cx45 protein abundance measured by immunoblotting was not different in Cx43-deficient or null hearts compared to wild-type control hearts. However, the amount and distribution of Cx45 immunoreactive signal measured by quantitative confocal analysis were markedly reduced in both Cx43(+/-) and Cx43( /-) hearts. CONCLUSION: Although the total content of Cx45 is not upregulated in Cx43-deficient hearts, the localization of Cx45 to cardiac gap junctions depends on the expression level of Cx43 and is dramatically altered in mice that express no Cx43. PMID- 11922903 TI - Bcl-xl reduces doxorubicin-induced myocardial damage but fails to control cardiac gene downregulation. AB - OBJECTIVE: We recently reported that doxorubicin (Dox), an effective anti-cancer drug, induces apoptosis in cardiac myocytes in association with reduction of Bcl xl expression. In the present study, we further examined whether overexpression of Bcl-xl ameliorates Dox-induced cardiac myocyte damage. METHODS AND RESULTS: Overexpression of the Bcl-xl gene by adenovirus vector resulted in an 11-fold increase in Bcl-xl protein in neonatal rat cardiac myocytes (BCL) compared to that in cells with beta-galactosidase gene transfection (CTL). Although Dox treatment generated similar amounts of reactive oxygen species (ROS) in BCL and CTL, cell viability was maintained and the number of apoptotic cardiac myocytes was significantly decreased in BCL. Cytochrome c release and enhanced caspase-3 activity after Dox treatment were significantly suppressed and Bax expression level was decreased in BCL. Cardiac-specific gene expression is known to be inhibited by Dox. The expression of cardiac alpha-actin and sarcoplasmic reticulum Ca(2+)-ATPase 2a mRNA was equally inhibited in BCL and CTL after Dox treatment. CONCLUSIONS: Overexpression of Bcl-xl in cardiac myocytes failed to regulate Dox-induced ROS generation and cardiac-specific gene downregulation but inhibited apoptosis accompanied by reduction of Bax protein. PMID- 11922904 TI - Significant prolongation of atrial monophasic action potential duration: short term reverse electrophysiological changes after internal cardioversion of atrial fibrillation. AB - OBJECTIVE: Since short action potentials and short refractory periods facilitate the induction of atrial reentry, this maladaptation has been proposed as the pathophysiological basis of the frequent immediate recurrences of atrial fibrillation (IRAF) after internal cardioversion. However, short-term reverse electrophysiological changes of the atria after cardioversion have not been studied in humans. METHODS: Thirty-seven patients with chronic atrial fibrillation of 16+/-19 months and ten patients with an atrial fibrillation duration < or =48 h underwent internal cardioversion. Antiarrhythmic medication was only continued in 10 patients (21%), who were on amiodarone before cardioversion. Atrial monophasic action potential duration at 90% repolarization (APD(90)), sinus rate, P wave duration and interatrial conduction times between high right atrium and coronary sinus were recorded at min 0, 1, 3, 5, 10, 15 and 20 after cardioversion. RESULTS: Internal cardioversion was successful in all patients, but twelve of the patients with chronic AF (32%) and three of the patients with intermittent AF (30%) had one to four episodes of IRAF after 16+/ 28 s. There was a significant 52+/-30 ms APD(90) prolongation, 83% of which occurred in min 0-3 (P<0.0001) and 17% in min 3-20 (P<0.05) after internal cardioversion. There was no significant temporal change in sinus rate, P-wave and interatrial conduction time during the time studied. APD(90) prolongation and its time dependence did not show a detectable difference in subgroups with chronic AF, IRAF, left atrial size >40 mm and treatment with amiodarone. CONCLUSIONS: There is a significant prolongation of action potential duration in min 0-3 after internal cardioversion of atrial fibrillation, whereas sinus rate and intra- and interatrial conduction time remain unchanged. APD(90) prolongation in min 0-3 shows a temporal relationship to the high rate of immediate recurrences of atrial fibrillation during this time interval. The data imply that there is a transient recovery of atrial refractoriness after cardioversion and suggest a mechanism of the high rate of early recurrences of atrial fibrillation. PMID- 11922905 TI - Vitronectin is up-regulated after vascular injury and vitronectin blockade prevents neointima formation. AB - OBJECTIVE: Smooth muscle cell (SMC) migration involves interactions with extracellular matrix (ECM) and is an important process in response to arterial wall injury. We investigated the expression and the functional role of vitronectin (VN) in the response after vascular injury. METHODS: VN and alpha v beta 3/beta 5 integrin expressions were investigated after balloon carotid injury of Sprague-Dawley rats. Adventitial delivery of blocking antibodies to VN, alpha v beta 5 and beta 3 integrins were performed to assess their roles in neointima formation. In vitro, migration assays were carried out on human SMC. RESULTS: Immunohistochemistry and in situ hybridization for VN showed an upregulation of VN during the early time points of intima formation. alpha v beta 3/beta 5 integrins expression correlated with VN expression. After 7 days, blocking antibodies to VN, alpha v beta 5 and beta 3 induced a significant decrease on intimal area associated with a decrease in intimal cell counts. A slight decrease in intimal cell proliferation without any effect on apoptosis was observed after VN blockade. In vitro, migrating SMC strongly expressed VN after injury and neutralizing anti-VN antibody inhibited SMC migration. Blocking experiment with anti-alpha v beta 5 and -alpha v beta 3 integrin antibodies showed that not only VN-alpha v beta 3 but also VN-alpha v beta 5 interactions are required for SMC migration. CONCLUSION: This study characterizes the VN-ECM interaction in SMC and supports the role of VN in mediating SMC migration and neointimal formation in response to injury. PMID- 11922906 TI - Chronic endothelin receptor blockade prevents renal vasoconstriction and sodium retention in rats with chronic heart failure. AB - OBJECTIVE: Importance of endothelin in mediating the chronic renal alterations of chronic heart failure was studied in rats chronically treated with bosentan after myocardial infarction. METHODS: Rats were subjected to coronary artery ligation and were treated for 8 weeks with placebo or bosentan, a dual ET(A) and ET(B) receptor antagonist, (approximately 100 mg/kg/day) as food admix. Sham-operated rats served as normal controls. Cardiac and renal functions were measured at the end of 8-week treatment. RESULTS: Bosentan significantly reduced the elevated left ventricular end-diastolic pressure (from 26.6+/-3.3 to 11.4+/-2.2 mmHg, P<0.001) and the increased heart-to-body-weight ratio seen in untreated rats with myocardial infarction. Bosentan prevented the marked increase in renal vascular resistance (bosentan, 7.7+/-0.6; untreated, 15.6+/-2.5 mmHg/ml/min; P<0.001). This led to a significant increase in renal plasma flow resulting in a decrease in filtration fraction. Bosentan furthermore increased urinary sodium excretion. CONCLUSIONS: Prolonged ET receptor blockade in rats with myocardial infarction has chronic renal vasodilatory effect and improves renal sodium excretory function. Thus, dual ET antagonists such as bosentan might be useful in the treatment of the progressive renal failure associated with human chronic heart failure. PMID- 11922907 TI - Expression of latent TGF-beta binding proteins and association with TGF-beta 1 and fibrillin-1 following arterial injury. AB - OBJECTIVES: Transforming growth factor-beta (TGF-beta), a potent regulator of wound healing and scar formation, is thought to have a key role in the response to arterial injury. Latent TGF-beta binding proteins (LTBPs), members of the fibrillin superfamily, govern TGF-beta1 release, targeting and activation in vitro and also play a role as structural components of fibrillin-rich microfibrils. Despite the potential of LTBPs to modulate the response to arterial injury through either or both of these mechanisms, as yet their expression and function in the injured vasculature remain poorly defined. METHODS: In this study, a porcine model of coronary angioplasty was used to investigate LTBP-1 and LTBP-2 synthesis and their association with TGF-beta 1 and fibrillin-1. RESULTS: After angioplasty, increased LTBP-1 and LTBP-2 immunostaining was detected in a similar distribution to increased TGF-beta 1 expression in the neointima and in the neoadventitia. Overnight organ cultures revealed the formation of large latent TGF-beta 1 complexes containing LTBP-1. Increased LTBP-1 proteolysis after arterial injury correlated with increased active and latent TGF-beta levels. LTBP 2 synthesis increased in response to arterial injury but was neither present in large latent complexes nor proteolytically processed. LTBP-1 and LTBP-2 both co localised to fibrillin-rich fibrillar structures in the neointima and adventitia. CONCLUSIONS: These data suggest that LTBP-1 may have a TGF-beta 1 binding role in the arterial response to injury, and that LTBP-1 and LTBP-2 may have a structural role in association with microfibrils within the developing neointimal lesion. LTBP-1 proteolysis is potentially an important regulatory step for TGF-beta activation in the vasculature and inhibition of proteolysis could represent a novel therapeutic modality for controlling the arterial injury response. PMID- 11922908 TI - Thrombostatin, a bradykinin metabolite, reduces platelet activation in a model of arterial wall injury. AB - OBJECTIVE: Thrombin activates platelets and contributes to the occlusion of arteries following thrombolytic therapy or angioplasty. Thrombostatin (RPPGF), the angiotensin converting enzyme degradation product of bradykinin, inhibits alpha-thrombin induced platelet activation. We hypothesized that thrombostatin prevents platelet aggregation and adhesion after balloon angioplasty (BA). METHODS: Platelet-rich plasma (PRP) was obtained from 22 Beagle dogs before sacrifice and 10% of the PRP was labeled with 111In. Carotid arteries were then removed from each dog and mounted in a dual perfusion chamber and intimal injury was performed with BA. 111In-PRP with or without thrombostatin or aspirin alone was perfused through the arteries for 60 min. During perfusion, platelet volume was measured using a Coulter counter and a laser-light scattering technique. Platelet adhesion to arteries was measured by radioactivity count. RESULTS: Arterial injury alone compared to non-injury increased platelet volume in the circuit by 1.4 times (x) (P<0.05) using a Coulter counter or 1.8x (P<0.05) using laser-light scattering and increased platelet adhesion by 2.3x (P<0.01). When compared to BA injury alone, the addition of thrombostatin reduced platelet volume by 1.8x (P<0.03) as measured by Coulter counter or 1.9x (P<0.01) by laser light scattering and platelet adhesion by 4.2x (P<0.05). Compared to BA injury alone, aspirin reduced platelet volume by 1.2x (P<0.01) as assessed by Coulter counter or 1.5x (P<0.03) using laser-light scattering and platelet adhesion by 1.8x (P<0.02). CONCLUSION: Thrombostatin or aspirin independently decreases evidence of platelet activation in the canine carotid artery model of BA injury. PMID- 11922909 TI - AAV-mediated VEGF gene transfer into skeletal muscle stimulates angiogenesis and improves blood flow in a rat hindlimb ischemia model. AB - OBJECTIVES: Clinical trials on therapeutic angiogenesis using vascular endothelial growth factor (VEGF) are ongoing, however the benefits of these therapies are still controversial. To establish a more efficient gene transfer method for ischemic diseases, we investigated the therapeutic potential of adeno associated virus (AAV)-mediated VEGF gene transfer. METHODS: We produced VEGF(165)-expressing AAV vectors (AAV-VEGF). HEK-293 cells were transduced with AAV-VEGF in vitro and VEGF expression and secretion were examined. We used a rat ischemic hindlimb model and AAV-VEGF was administered intramuscularly into the ischemic limb. Gene expression was evaluated by RT-PCR and ELISA. Six weeks after gene transfer, we measured the blood flow of limb vessels and the skin temperature of limbs. Histochemical examination was performed to illustrate capillary growth. RESULTS: Western blotting and ELISA revealed VEGF protein expression and secretion from AAV-VEGF-transduced HEK-293 cells. VEGF mRNA and protein expression was consistently observed in the injected muscle at least 10 weeks after the injection, while no VEGF mRNA could be detected at remote organs. The mean blood flow in AAV-VEGF-transduced ischemic limbs was significantly higher than in AAV-LacZ-transduced limbs. Capillary density was significantly higher in AAV-VEGF-injected tissues than in AAV-LacZ-injected tissues. CONCLUSIONS: This study demonstrates that (1) AAV-mediated VEGF gene transfer into rat skeletal muscles is efficient and stable without ectopic expression, and (2) AAV-mediated VEGF gene transfer stimulates angiogenesis and thereby improves blood flow in a rat hindlimb ischemia model. These findings suggest that AAV mediated VEGF gene transfer may be useful for treatment of ischemic diseases. PMID- 11922910 TI - The atherogen 3-methylcholanthrene induces multiple DNA adducts in mouse aortic smooth muscle cells: role of cytochrome P4501B1. AB - OBJECTIVE: 3-Methylcholanthrene (MC), a polycylic aromatic hydrocarbon, induces atherogenesis in mice fed an atherogenic diet. In this study, we tested the hypothesis that MC would induce DNA adducts in mouse aortic smooth muscle cells (SMCs) and that cytochrome P4501B1 (CYP1B1) plays an important role in the activation of MC to genotoxic intermediates. METHODS: Cultured SMCs were treated with MC or the vehicle dimethyl sulfoxide (DMSO), and DNA was isolated after 24 h. In some experiments, the cells were pre-treated with the CYP1B1 inhibitor 1 ethynylpyrene (EP) prior to exposure to MC. DNA adducts were determined by the 32P-postlabeling assay. Aryl hydrocarbon hydroxylase assay was measured by fluorimetry. RESULTS: MC induced formation of 12 DNA adducts that were not observed in DMSO-treated cells. DNA adduct formation was dose-dependent, with maximum response observed at 3 microM. Pre-treatment of cells with EP dramatically suppressed DNA adduct formation by MC. MC treatment caused induction of CYP1B1, but not CYP1A1. CONCLUSION: The induction of high levels of multiple DNA adducts in SMCs by MC suggests that SMCs have a functional enzymatic machinery capable of metabolically activating MC to genotoxic metabolites. The significant inhibition by EP of MC-induced DNA adduct formation indicated that CYP1B1 was the primary CYP enzyme responsible for formation of genotoxic metabolites that may play a role in the induction of atherosclerosis by MC. PMID- 11922911 TI - Chronic antioxidant supplementation attenuates nuclear factor-kappa B activation and preserves endothelial function in hypercholesterolemic pigs. AB - OBJECTIVE: Hypercholesterolemia (HC), a pro-oxidant condition, activates nuclear factor-kappa beta (NF-kappa B) and is associated with coronary endothelial dysfunction. The physiological significance of in vivo chronic antioxidant intervention on HC-induced NF-kappa B activation and coronary endothelial function remains unclear. METHODS: Four groups of pigs were studied after 12 weeks of normal diet, normal diet with concomitant antioxidant intervention (100 IU/kg of vitamin E and 1 g of vitamin C daily), 2% HC diet, or HC diet+antioxidant supplementation. NF-kappa B activation and the nitric oxide (NO) pathway were investigated by Western blotting and immunohistochemistry, while oxidative stress was evaluated by coronary artery tissue radical scavenger activity and levels of vitamin E and C. Endothelial function was studied in vitro by coronary vasoreactivity to bradykinin and substance P. RESULTS: HC animals had increased activation of NF-kappa B, decreased endothelial NO synthase expression, and decreased radical scavenger system activity, associated with impaired coronary endothelial function. Antioxidant supplementation in HC normalized NF kappa B activation and NO bioactivity, and preserves coronary endothelial function. CONCLUSIONS: This study demonstrates for the first time that in vivo chronic interruption of the endogenous oxidative stress cascade reduces HC induced NF-kappa B activation and normalizes NO bioactivity in association with preservation of coronary endothelial function. This study suggests a role for increased oxidative stress and NF-kappa B activation in early atherosclerosis. PMID- 11922912 TI - Reversal of chronic hypoxia-induced alterations in pulmonary artery smooth muscle electromechanical coupling upon air breathing. AB - OBJECTIVE: Chronic hypoxia (CH) induces selective pulmonary hypertension which is accompanied by structural and functional alterations in the pulmonary vasculature. Little information is available on the regression of CH-induced functional alterations of pulmonary wall. In the present work, we investigated the reversal of CH-induced pulmonary hypertension with a special focus on alterations in the electrophysiological properties of pulmonary artery smooth muscle cells (PAMCs) after normoxia recovery. METHODS: Rats were exposed to a hypobaric environment for 3 weeks (CH rats) and then subjected to a normoxic environment for 3 weeks (normoxia-recovery group) and compared with rats maintained in a normoxic environment (control rats). Electrophysiological properties of PAMCs were studied using conventional microelectrodes and patch clamp technique. RESULTS: CH rats exhibited a threefold increase in pulmonary blood pressure compared to control rats and this increase was fully reversed following 3 weeks of normoxia. PAMCs from CH rats were depolarised (about 20 mV), had an elevated calcium concentration and exhibited a hypersensitivity to 4 aminopyridine (4-AP) of membrane potential as well as the tone of arterial rings compared with tissues from control rats. Whole cell patch-clamp recordings indicated that voltage gated potassium channel currents I(Kv) and I(K(N)) were decreased in PAMCs from CH rats with a hyper sensitivity of I(K(N)) to 4-AP. CH induced alterations in electrophysiological properties of PAMCs were also fully reversed after 3 weeks of normoxia recovery. CONCLUSIONS: Both the increase in the pulmonary blood pressure and alterations in electrophysiological properties of PASMCs simultaneously reverse after normoxia recovery. This complete reversibility of all of the CH-induced pulmonary vascular alterations suggests that curative treatments for PAHT may now be designed aimed at targeting the very limited key factors implicated in hypoxia sensing. PMID- 11922913 TI - Genotype at a promoter polymorphism of the interleukin-6 gene is associated with baseline levels of plasma C-reactive protein. AB - OBJECTIVE: Baseline concentrations of plasma C-reactive protein (CRP) are associated with coronary heart disease. Interleukin-6 (IL-6) regulates CRP gene expression; a promoter polymorphism (-174G/C) of the IL-6 gene has been shown to influence IL-6 transcription but the relationship between genotype at this polymorphism and circulating levels of inflammatory markers remains unclear. We hypothesised that plasma CRP would be a heritable phenotype that would be influenced by genotype at this polymorphism. METHODS: We measured baseline plasma CRP and determined genotypes at the -174G/C polymorphism of the IL-6 gene in 588 members of 98 nuclear families. The heritability of plasma CRP and the association of plasma CRP with genotype were determined using variance components methods. RESULTS: Baseline CRP levels were highly heritable (h(2)=0.39, P<0.0000001). Presence of the -174C allele was associated with higher baseline CRP levels, both in the whole population (P=0.01), and in the founders only (n=128, P=0.001). Family-based analyses confirmed the association (P=0.02) suggesting that it arises from chromosomal proximity or identity of the typed polymorphism with a genetic variant influencing baseline CRP levels. CONCLUSIONS: Baseline plasma CRP is a significantly heritable cardiovascular risk factor. Levels are associated with genotype at the -174G/C polymorphism of the IL-6 gene. PMID- 11922914 TI - Oxidant stress mechanism of homocysteine potentiating Con A-induced proliferation in murine splenic T lymphocytes. AB - OBJECTIVE: An elevated plasma homocysteine (Hcy) level is considered an independent risk factor for atherosclerosis. However, the mechanisms by which hyperhomocysteinemia induces atherosclerosis are only partially understood. The effect of Hcy on T lymphocyte proliferation and its mechanisms were examined in normal and hyperhomocysteinemia ApoE-knockout mice. METHODS: The mouse splenic T cells were treated with Hcy, related compounds and/or antioxidants in the presence or absence of Concanavalin A (Con A). DNA synthesis, cell apoptosis, interleukin-2 level and production of reactive oxygen species (ROS) were measured. RESULTS: Hcy (0.3-3.0 mM) and related compounds with thiol (-SH), such as cysteine and glutathione significantly potentiated Con A-induced proliferation and partially inhibited apoptosis in T lymphocytes, but it had no direct effect on resting T lymphocyte. ApoE-knockout mice with hyperhomocysteinemia (the level of plasma Hcy was 20.3+/-2.9 vs. 2.6+/-0.6 microM in control group, P<0.05) had a significant promotion of T-cell proliferation in response to Con A. Hcy (0.3-3.0 mM) also increased the intracellular ROS. Radical scavengers reduced Hcy effect. CONCLUSIONS: These data indicate that ROS generated by thiol (-SH) of Hcy auto oxidation are involved in Hcy effect on Con A-induced T lymphocyte proliferation. These findings suggest a novel mechanism may be involved in chronic inflammatory progression of atherosclerosis with hyperhomocysteinemia. PMID- 11922915 TI - Re: The sinoatrial node, connexin distribution patterns and specific immunodetection of connexin45. PMID- 11922917 TI - Epitope analysis of human immunoglobulin D with a mouse monoclonal anti-Fab delta chain antibody. AB - BACKGROUND: The clinical significance of IgD measurements in serum is limited as the frequency of abnormal concentrations is rare. METHODS: We prepared a mouse monoclonal antibody and a rabbit polyclonal antibody against Fab delta and Fc delta chain and compared epitope recognition by the monoclonal antibody against Fab delta (anti-Fab delta mono) with that by other antibodies. RESULTS: Anti-Fab delta mono specifically reacted with purified IgD and Fab delta of myelomatous origin, but not with other isotypes and light chains. In 19 of 22 myeloma sera, the monoclonal antibody recognized intact IgD and/or its fragments when analyzed by immunoblotting. Of these there were only four cases in which possible Fab delta fragments were identified. The other three sera showed no reactivity with the antibody and the IgD value was low on a chemiluminescence enzyme immunoassay. CONCLUSIONS: These findings indicate the presence of at least two immunochemically different IgD molecules in the sera. No positive reaction with any synthetic peptide was observed for the antibody on delta-chain ranging from N terminus of JH, C delta 1 to a hinge region. We suggest that the epitope recognized by the antibody is related to the variable region or a conformational structure on the Fd delta region of IgD. PMID- 11922918 TI - Is the neutrophil reactive oxygen species production measured by luminol and lucigenin chemiluminescence intra or extracellular? Comparison with DCFH-DA flow cytometry and cytochrome c reduction. AB - BACKGROUND: Polymorphonuclear neutrophils (PMNs) are crucial in host defense against invading microorganisms through reactive oxygen species (ROS) production. However, generated ROS released in excess into media can damage the host tissue. It is therefore essential, when exploring oxygen species production, to discriminate between its intracellular (IC) and extracellular (EC) localization. Several methods of ROS detection are commonly used. However, the literature shows that it is not always clear whether the species detected are IC or EC, especially with the chemiluminescence technique. METHODS: We compared PMN ROS production, determined by chemiluminescence, using two different probes (luminol and lucigenin) with that measured by 2'-7'-dichlorofluorescin diacetate (DCFH-DA) flow cytometry for IC production and by cytochrome c reduction for EC production. RESULTS: We found that luminol-dependent chemiluminescence explored IC ROS production more specifically (r=0.77, p<0.01: correlation between luminol amplified chemiluminescence and DCFH-DA flow cytometry). Lucigenin-amplified chemiluminescence and cytochrome c reduction were closely related (r=0.55, p<0.01). CONCLUSION: Luminometry detection can thus afford reproducible information on intracellular ROS kinetic production using luminol and extracellular ROS detection using lucigenin, simply and at low cost. PMID- 11922919 TI - Elevated soluble CD40 ligand is related to the endothelial adhesion molecules in patients with acute coronary syndrome. AB - BACKGROUND: Increasing evidence indicates that the CD40-CD40L interaction plays a pivotal role in the inflammatory regulation of atherosclerosis. Adhesion molecules especially the vascular adhesion molecules also play an important role in the pathogenesis of atherosclerosis which act as markers of inflammation. These inflammatory factors render vulnerability to the atherosclerotic plaque by triggering the fissure, rupture, and subsequent thrombosis, leading to the clinical scenario of unstable angina and acute myocardial infarction. METHODS: The difference of sCD40L concentration in different subtype of coronary heart disease and its relationship with vascular adhesion molecules was investigated. Enzyme-linked Immunosorbent Assay (EIA) was used to measure the serum sCD40L, soluble intercellular adhesion molecule-1 (sICAM-1), and soluble vascular cell adhesion molecule-1 (sVCAM-1). RESULTS: The sCD40L concentration was significantly higher in patients with acute coronary syndrome (ACS) (3.17+/-2.84 ng/ml) than in controls (1.19+/-1.05 ng/ml, p<0.01) and in patients with stable coronary heart disease (1.61+/-1.46 ng/ml, p<0.05). The sCD40L concentration was positively correlated with sICAM-1 (r=0.413, p<0.01), triglycerides (TG) (r=0.23, p<0.05), apoB (r=0.248, p<0.05), and HDL-cholesterol (r=-0.253, p<0.05). CONCLUSIONS: The sCD40L concentration was increased in acute coronary syndrome, suggesting the possible relation of CD40L to the pathogenesis. The serum CD40L concentration was positively correlated with adhesion molecule and was negatively associated with serum high-density lipoprotein cholesterol (HDL-C). PMID- 11922920 TI - Plasma alpha2-HS glycoprotein concentrations in patients with acute myocardial infarction quantified by a modified ELISA. AB - BACKGROUND: Human alpha2-HS glycoprotein (alpha2-HSG) is synthesized and secreted by the liver into circulation. Plasma concentrations of alpha2-HSG decrease significantly following infection, inflammation and malignancy. Since increased plasma concentrations of C-reactive protein are observed in patients with acute myocardial infarction (AMI), we hypothesized that plasma concentrations of alpha2 HSG would decrease during the initial phase of AMI and begin to increase in the recovery phase. METHODS: Twenty patients diagnosed with AMI were recruited for the study. A sensitive and specific ELISA was developed to assay alpha2-HSG concentrations in plasma. RESULTS: In AMI patients, plasma alpha2-HSG concentrations were decreased (281.3+/-25.8 mg/l, ranging from 132 to 489 mg/l on admission) compared to healthy individuals (312.3+/-9.9 mg/l, ranging from 210 to 450 mg/l) (P= 0.142). Interestingly, 40% of AMI patients demonstrated alpha2-HSG concentrations below 200 mg/l compared to none in the healthy control group. During the recovery period, alpha2-HSG concentrations begin to increase, with a mean+/-SEM of 290.1+/-22.1 mg/l. Regression analysis comparing plasma alpha2-HSG concentrations on admission to concentrations on discharge showed a significant positive correlation in matched-pair patient samples (P<0.01, r=0.45). CONCLUSIONS: We conclude that, in contrast to C-reactive protein, alpha2-HSG functions as a negative acute phase protein in AMI patients. Plasma alpha2-HSG concentrations start to decrease within a few hours after the onset of AMI and return to near normal concentrations during the recovery period (5-7 days after AMI). PMID- 11922921 TI - Tongkyutang inhibits mast cell-dependent allergic reactions and inflammatory cytokines secretion. AB - BACKGROUND: Tongkyutang (TKT) is an Oriental herbal prescription, which has been successfully applied for the treatment of allergic disorders, mainly allergic rhinitis in clinical medicine. However, its effect in experimental models remains unknown. METHODS: In a mouse model, the role of TKT was examined in mast cell dependent allergic reactions and secretion of inflammatory cytokines. RESULTS: TKT concentration-dependently inhibited the ear-swelling response induced by intradermal injection of compound 48/80. TKT inhibited the compound 48/80-induced degranulation from mast cells in ear tissue. TKT dose-dependently inhibited the histamine release from the rat peritoneal mast cells by compound 48/80. TKT also showed inhibition of anti-dinitrophenyl IgE antibody-induced passive cutaneous anaphylaxis reaction by oral administration. Furthermore, TKT inhibited both IL 1beta and TNF-alpha secretion induced by PMA and A23187, respectively. CONCLUSIONS: Our findings provide evidence that TKT inhibits the mast cell dependent allergic reactions and inflammatory cytokines secretion. PMID- 11922922 TI - Evaluation of the Wampole Laboratories ELISA-based assay for Epstein-Barr virus serology. AB - BACKGROUND: Epstein-Barr virus (EBV) infection is associated with infectious mononucleosis, Burkitt's lymphoma, and nasopharyngeal carcinoma. Serologic diagnosis of acute EBV infections has been the method of choice, and tests are available as indirect fluorescent antibody (IFA)- and ELISA-based assays. OBJECTIVE: To evaluate the ELISA-based EBV assay from Wampole Laboratories (Cranbury, NJ). METHODS: One hundred fifty-two consecutive samples received for comprehensive EBV serology were analyzed. RESULTS: A comparison of the Wampole Laboratories' ELISA system with the Gull/Meridian Diagnostics (Cincinnati, OH) IFA, and ELISA assays showed 88% concordance for anti-viral capsid antigen (VCA) IgM (n=177); 79% concordance for anti-VCA IgG (n=177); 87% concordance for anti NA IgG (n=172); and 48% concordance for anti-EA IgG (n=165). Using the results from all four antibody assays to identify patients with acute infection, the Wampole system had a 67% concordance with the Gull/Meridian (n=164). CONCLUSIONS: Differences in the specificity of the anti-EA IgG assays (i.e. reactivity against the D component of early antigen (EA-D) (Wampole) vs. reactivity to EA-D and the R component of early antigen (EA-R) (Gull/Meridian)) may have lead to poor concordance (48%) for this particular assay. Because the Wampole system had a <70% overall concordance with the Gull/Meridian system in diagnosis of acute infection, these data suggest that further studies are needed to determine the true clinical sensitivity and specificity of this system. PMID- 11922923 TI - The effect of different buffers and amounts of intestinal alkaline phosphatase isoforms on total alkaline phosphatase activity. AB - BACKGROUND: The transphosphorylating accepter buffers (2-amino-2-methyl-1 propanol, AMP; N-methyl-D-glucamine, MEG; diethanolamine, DEA and 2 ethylaminoethanol, EAE) have been widely used for the measurement of serum total alkaline phosphatase activity (ALP) in clinical laboratories, and the individual isozyme are activated differently by respective buffers. MATERIALS AND METHODS: We examined the activity of serum ALP using four buffers with levels of both high molecular weight intestinal alkaline phosphatase (HIAP) and normal molecular weight intestinal alkaline phosphatase (NIAP). We classified 80 healthy subjects into two groups of blood group B or O secretors (n=36) and other blood groups (n=44). RESULTS: The mean ALP activities at fasting in blood group B or O secretors from AMP, MEG, DEA and EAE methods were 15.5%, 24.0%, 11.0% and 22.1% higher than those in other blood groups, respectively. The reference ranges of ALP activity at fasting with the AMP method in blood group B or O secretors and other blood groups were 63.5+/-17.4 U/l (mean+/-S.D.) and 55.0+/-14.5 U/l (mean+/ S.D.), respectively. The difference between the reference ranges of ALP activity in blood group B or O secretors and other blood groups was statistically significant (p<0.01). HIAP and NIAP in serum at fasting only appeared in blood group B or O secretors, and the activities of HIAP and NIAP were 4.7+/-3.4 U/l (mean+/-S.D.) and 2.2+/-1.2 U/l (mean+/-S.D.), respectively. The activity of ALP (HIAP+NIAP) in blood group B or O secretors was 56.6+/-15.1 U/l (mean+/-S.D.), and this reference range was approximately the same as the ALP activity (55.0+/ 14.5 U/l) of other blood groups. The same results were observed with MEG, DEA and EAE methods. CONCLUSIONS: These results suggested that the differences in ALP activity in blood group B or O secretors and other blood groups were closely related to the HIAP and NIAP levels. PMID- 11922924 TI - Assessing nitrate metabolism in the intestinal tract by measuring breath nitric oxide and nitrous oxide, and its clinical significance. AB - BACKGROUND: The toxicity of dietary nitrate (NO3-) is controversial. One reason is nitrate metabolism in the intestine is so complicated that it is far from fully understood. There is no study measuring breath nitric oxide (NO) and nitrous oxide (N2O) after ingesting vegetables and high-nitrate food at the same time. METHODS: Breath samples from 10 healthy young and 10 healthy old subjects were collected at 15-min intervals for 5 h after ingestion of 100 g of lettuce and during fasting (control). Breath NO and N2O were analyzed by a chemiluminescence and an IR-PAS analyzer respectively. RESULTS: N2O maximum concentration and excretions increased significantly after ingesting lettuce in each group [303 (30) vs. 750 (81) ppb, 771 (72) vs. 1668 (146) microg in young; 442 (52) vs. 1092 (109) ppb, 1088 (125) vs. 2100 (183) microg in old subjects; mean (SE), P<0.01], while NO did not. In addition, breath NO was strongly influenced by ambient NO, which varied greatly. N2O maximum level in old subjects after ingesting lettuce was higher than that of young subjects (750 vs. 1092 ppb, P<0.05), and significantly higher N2O concentration levels were seen at 30, 45, 60, and 105 min in old subjects. CONCLUSIONS: A large amount of N2O produced in the intestine and normal nitrate intake do not influence the breath NO concentration, probably due to its relatively small production. Higher maximum N2O concentration after ingesting lettuce in old subject is probably because more bacteria, which rapidly reduce dietary nitrate in the upper intestinal tract, inhabit the gut in old age. Our results suggested that breath N2O is a useful noninvasive maker to estimate dietary nitrate reduction in the intestinal tract. PMID- 11922925 TI - Stability of total plasma homocysteine in perinatology. AB - BACKGROUND: Typical assay methods for total homocysteine in human plasma involve EDTA-containing whole blood. Unfortunately, rapid increases of the plasma homocysteine concentrations due to cellular export are observed when the EDTA containing samples are not stored on ice and processed shortly after collection. This is a cumbersome procedure in perinatal settings, whereby delivery usually takes place at unpredictable times. METHODS: The stability of homocysteine was assessed from six placental and neonatal blood samples collected in citrate buffer. Samples were stored at 4 degrees C and tested at regular intervals for the first 24 h. RESULTS: There was no statistical difference in homocysteine concentrations as observed over the study storage period (mean coefficient of variation [CV] 4.9%). CONCLUSIONS: Citrated samples can be left in a refrigerator for at least 24 h with no effect on the plasma homocysteine concentrations. PMID- 11922927 TI - Mercurimetric determination of chloride in sweat. PMID- 11922926 TI - Quantitative RT-PCR analysis demonstrates that synthesis of the recombinant fibrinogen is dependent on the transcription and synthesis of gamma-chain. AB - BACKGROUND: The purpose of this study was to examine the relationship between the production of secreted fibrinogen and the synthesis of gamma-chain mRNA. METHODS: We transfected a gamma-chain expression vector into Chinese hamster ovary cells already expressing both Aalpha- and Bbeta-chains of fibrinogen and measured fibrinogen output concentrations by ELISA. We quantified both gamma-chain and Bbeta-chain mRNA concentrations using the recently developed TaqMan fluorogenic detection system. RESULTS: The concentration of secreted fibrinogen into the media positively correlated with the amount of fibrinogen contained in the cell lysates. Additionally, quantitative mRNA assays revealed that the fibrinogen concentration in the cell lysates correlated well with the concentration of gamma chain mRNA (r=0.7077, p<0.01) but not with the concentration of Bbeta-chain mRNA (r=0.0224, NS). CONCLUSIONS: These results demonstrate that the amount of recombinant fibrinogen produced in cells transfected with the gamma-chain vector, also expressing normal Aalpha- and Bbeta-chains, is dependent on the transcription of gamma-chain mRNA. Namely, in this recombinant expression system using a two-step transfection procedure, gamma-chain synthesis is the rate limiting factor for fibrinogen production. This quantitative method to measure mRNA may prove very useful for further in vivo analysis of fibrinogen gene transcription. PMID- 11922929 TI - Evaluation: salvation or nemesis of medical informatics? AB - The currently prevailing paradigms of evaluation in medical/health informatics are reviewed. Some problems with application of the objectivist approach to the evaluation of real-rather than simulated-(health) information systems are identified. The rigorous application of the objectivist approach, which was developed for laboratory experiments, is difficult to adapt to the evaluation of information systems in a practical real-world environment because such systems tend to be complex, changing rapidly over time, and often existing in a variety of variants. Practical and epistemological reasons for the consequent shortcomings of the objectivist approach are detailed. It is argued that insistence on the application of the objectivist principles to real information systems may hamper rather than advance insights and progress because of this. Alternatives in the form of the subjectivist approach and extensions to both the objectivist and subjectivist approaches that circumvent the identified problems are summarized. The need to include systems engineering approaches in, and to further extend, the evaluation methodology is pointed out. PMID- 11922931 TI - Evaluation in the design of health information systems: application of approaches emerging from usability engineering. AB - This paper examines the role of evaluation in the design of health care information systems. A framework is presented for considering evaluation in the context of software development processes, in particular, the systems development life cycle (SDLC). Variations on standard design methodologies are then discussed, including methods based on rapid development and continual evaluation of prototype systems. Usability testing is presented as a key method for conducting evaluations during iterative system development. The emergence of design methodologies, where evaluation is viewed as a central part of the development cycle is also discussed. Evaluation methodologies are then considered along a continuum, ranging from studies involving a high degree of experimental control to observational approaches. A full cycle approach to evaluation of health care systems is argued for, involving deployment of new methods across the SDLC. Implications for future work exploring the integration of design and evaluation processes in health informatics are discussed. PMID- 11922930 TI - Improving system quality through software evaluation. AB - The role of evaluation is examined with respect to quality of software in healthcare. Of particular note is the failure of the Therac-25 radiation therapy machine. This example provides evidence of several types of defect which could have been detected and corrected using appropriate evaluation procedures. The field of software engineering has developed metrics and guidelines to assist in software evaluation but this example indicates that software evaluation must be extended beyond the formally defined interfaces of the software to its real-life operating context. PMID- 11922932 TI - Evaluation in health informatics: computer simulation. AB - The evaluation of complex medical informatics applications involves not only the information system, but also its impact on the organizational environment in which it is implemented. In instances where these applications cannot be evaluated with traditional experimental methods, computer simulation provides a flexible approach to evaluation. The construction of a computer simulation model involves the development of a model that represents important aspects of the system under evaluation. Once validated, the model can be used to study the effects of variation in system inputs, differences in initial conditions and changes in the structure of the system. Three examples are discussed, namely, a wide-area health care network, physician order entry into a hospital information system, and the use of an information system designed to prevent medical errors that lead to adverse drug events in hospitals. PMID- 11922933 TI - Beyond effectiveness: the evaluation of information systems using A Comprehensive Health Technology Assessment Framework. AB - A Comprehensive Health Technology Assessment Framework is presented as a conceptual tool for decision-making about health technologies, including information technologies. The aim of the model is to provide an empirical, evidence-based foundation for health technology decisions. The major framework dimensions are (1) population at risk, (2) population impact, (3) economic concerns, (4) social context (including ethical, legal, and political concerns), and (5) technology assessment information. This multi-disciplinary approach provides guidelines on use of appropriate information in aligning 'stakeholder wants' and 'population needs'. PMID- 11922935 TI - The TEAM methodology for the evaluation of information systems in biomedicine. AB - The TEAM evaluation methodology for information systems in biomedicine (Total Evaluation and Acceptance Methodology) is a unifying methodology for any computer based information system based on a three dimensional framework; these dimensions being Role, Time and Structure. The theory is derived from how the information system relates to the general system where it should operate, the properties of information flow within a general system and the relation between a system and its models. As a system can in theory be modelled from many perspectives, a perspective to be modelled is built up by formulating criteria relevant to that perspective which can be evaluated by quantitative and qualitative assessment methods. Key characteristics of the methodology include the insistence on a global rather than partial approach to the evaluation of information systems, also the dynamic nature of an information system which is continually in modification as it more successfully deals with the inherent complexity of the environment in which it is operating. The role dimension identifies four main categories, designer, specialist user, end user and stakeholder from which several sub-categories may be identified. The time dimension has four main phases towards relative stability of the information system. The structural dimension distinguishes strategic, tactical or organisational and operational levels that often are confused together with risk of dilution in current approaches. It is believed that this framework and methodology can provide a basis for future standardisation of evaluation methodologies. PMID- 11922934 TI - Evaluation in health informatics: social network analysis. AB - Social network analysis comprises a set of research methods that can be used to analyze the relationships among entities such as people, departments, and organizations. The purpose of the analysis is to discover patterns of relationships that affect both individual and organizational attitudes and behavior such as the adoption, diffusion, and use of new medical informatics applications. This paper presents an introduction to the concepts and methods of social network analysis. Several applications to health informatics are described. PMID- 11922936 TI - 'CHEATS': a generic information communication technology (ICT) evaluation framework. AB - This paper describes a generic framework for the evaluation of information communication technologies. This framework, CHEATS, utilises both qualitative and quantitative research methods and has proved appropriate in multiple clinical settings including telepsychiatry, teledermatology and teleeducation. The paper demonstrates how a multidisciplinary approach is essential when evaluating new and emerging technologies, particularly when such systems are implemented in real service as opposed to a research setting. PMID- 11922937 TI - A proposal to use a balanced scorecard to evaluate Information for Health: an information strategy for the modern NHS (1998-2005). AB - The author was invited to assist in the development of an evaluation methodology for the Strategy. One of the conundrums of measuring the information management & technology (IM&T) function is that infrastructure investments cannot be cost justified on a return on investment basis. The balanced scorecard (BSC) is a means to evaluate corporate performance from four different perspectives: the financial perspective, the internal business process perspective, the customer perspective, and the learning and growth perspective. An IM&T BSC for Information for Health was recommended as means of allowing managers to see the positive and negative impacts of IM&T activities on the factors that are important to the NHS as a whole. PMID- 11922938 TI - GPI-anchoring of GM-CSF results in active membrane-bound and partially shed cytokine. AB - Granulocyte-macrophage colony-stimulating factor (GM-CSF) can induce the generation and activation of dendritic cells (DCs), the most potent of antigen presenting cells (APCs). Tumors secreting GM-CSF have been shown to induce strong anti-tumor immune responses. In this report, we have constructed a glycosylphosphatidyl-inositol (GPI) anchored form of GM-CSF (GPI-GM-CSF). This protein subsequently was found expressed on the cell membrane and sensitive to phosphatidyl-inositol-specific phospholipase C (PIPLC), confirming that it is GPI anchored. However, GM-CSF was also found in the culture supernatant of cells expressing GPI-GM-CSF. Inhibition studies using brefeldin A and para-formaldehyde fixation revealed that GM-CSF found in the supernatant was not secreted, but due to shedding or proteolytic cleavage. Accumulation of GM-CSF in the media from isolated membranes was time and temperature-dependent. The released portion represented 10-15% of all membrane-bound GM-CSF after 72h under culture conditions. GPI-GM-CSF retained functional activity to induce bone marrow cell proliferation and administration of GPI-GM-CSF expressing membranes induced the generation of DCs in vivo. These results demonstrate that GPI-anchored GM-CSF retains all functional activity of native GM-CSF while gaining the ability to attach to cell membranes. The ability of GPI-GM-CSF to be expressed on membranes and be partially released, can possibly lead to formation of a cytokine gradient, while retaining ability to target associated membrane antigens to DCs. This novel form of GM-CSF may have wide range of clinical applicability. PMID- 11922939 TI - TREM-1, MDL-1, and DAP12 expression is associated with a mature stage of myeloid development. AB - The triggering receptor expressed on myeloid cells (TREM-1) and the myeloid DAP12 associating lectin (MDL-1) are two recently identified receptors which associate non-covalently with DAP12 to form receptor complexes involved in monocytic activation and inflammatory response. In this study, we investigated whether the expression of TREM-1, MDL-1, and DAP12 correlated with myelomonocytic differentiation. Northern and RT-PCR revealed a strong expression of TREM-1, MDL 1, and DAP12 in peripheral blood-derived CD14(+) mature monocytes in contrast to undifferentiated bone marrow CD34(+) stem cells, and in the differentiated versus undifferentiated U937 cells. TREM-1 and MDL-1 RNA expression was also more elevated in adult than fetal tissues and in normal than malignant cells. These findings suggest that the TREM-1/DAP12 and MDL-1/DAP12 signaling pathways are features of mature differentiated myelomonocytic cells. In addition, expression of an alternative mRNA TREM-1 splice variant (TREM-1sv) was detected that might translate into a soluble receptor with potential as a regulator of myeloid activation. PMID- 11922940 TI - Influence of terminal residue on adjacent disaccharide immunogenicity. AB - Aberrant O-glycosylation of cell surface mucin antigens is characteristic of epithelial cancer cells. For example, Thomsen-Friedenreich disaccharide (TFD) is a chemically well-defined carbohydrate antigen with a documented link to malignancy. There have been many attempts to improve immune response to carbohydrate antigens, for use in immunotherapy. As part of an alternative strategy to improve carbohydrate immunogenicity, we studied the influence of terminal benzyl (Bzl) or p-nitrophenyl (pNP) residue on immunogenicity of adjacent TFD. Mice immunized with keyhole limpets hemocyanin-TFD (KLH-TFD), KLH TFD(alpha)Bzl, or KLH-TFD(alpha)pNP produced anti-KLH antibodies, which were analyzed by enzyme-linked immunosorbent assay (ELISA). KLH-TFD did not give significant anti-TFD antibody titer, confirming the poor immunogenicity of TFD. Immunization with KLH-TFD(alpha)Bzl and KLH-TFD(alpha)pNP raised antibody titers against TFD(alpha)Bzl and TFD(alpha)pNP, respectively. KLH-TFD(alpha)Bzl also gave higher anti-TFD antibody response, whereas KLH-TFD(alpha)pNP did not, indicating that terminal Bzl residue improves immune response to adjacent carbohydrate. Analysis of anti-TFD(alpha)Bzl or anti-TFD(alpha)pNP IgG antibodies by competitive ELISA, using carbohydrate-related antigens as inhibitors, demonstrated their high specificity to their respective antigens. Anti TFD(alpha)pNP antibody was not inhibited by TFD, but was significantly inhibited by GalNAc(alpha)pNP. The fact that p-nitrophenol (pNPol) has more competitive ability that GalNAc indicates that terminal polar residue is the main target antigen. In contrast, anti-TFD(alpha)Bzl antibody was inhibited to a similar degree by GalNAc(alpha)Bzl and TFD, confirming the carbohydrate recognition by antibodies yielded by terminal non-polar modification of the immunogen. PMID- 11922941 TI - Antibody-mediated neutralization of cytomegalovirus: modulation of efficacy induced through the IgG constant region. AB - Antibodies can neutralize the infectious properties of human cytomegalovirus (CMV). In vivo, the major neutralization determinants are located on glycoprotein B (gB). Recombinant human antibodies, that carry different constant regions (IgG1, IgG3 and the synthetic variant IgG3mA) against two of these epitopes were investigated for their ability to recruit the complement cascade for destruction of the virus. It was shown that all variants of an antibody against the antigenic domain (AD)-2 epitope displayed a similar neutralization activity despite the fact that improved C1q binding was observed for IgG3 and IgG3mA over the IgG1 variant. In contrast, an antibody against the AD-1 epitope carrying the normal IgG3 constant region, was less efficient than its IgG1 counterpart in neutralizing the virus in the absence of complement. However, it restored its activity in the presence of complement to the level of the naturally occurring IgG1 version. The same antibody was substantially more potent in neutralizing the virus in the presence of complement if it carried the IgG3mA constant region. This demonstrates the importance of the constant domain for the biological activity of AD-1 specific antibodies, a factor that should be taken into account when using antibody-based therapeutics or when inducing antibodies by vaccination. PMID- 11922942 TI - T cell receptor excision circle assessment of thymopoiesis in aging mice. AB - Signal joint T cell receptor delta (TCRD) excision circles (TRECs) are episomal DNA circles generated by the DNA recombination process that is used by T lymphocytes to produce antigen-specific alpha/beta T cell receptors. Measurement of TRECs in thymocytes and peripheral blood T cells has been used to study thymus output in chickens and humans. We have developed a real-time quantitative-PCR assay for the specific detection and quantification of mouse TCRD episomal DNA circles excised from the TCRA locus during TCRA gene rearrangement (mTRECs). We found that the mouse TCRD TRECs detected with this assay were predominantly in naive phenotype CD4(+) and CD8(+) T cells. In a series of aged mice (range 6-90 week-old) we determined the absolute number of thymocytes and the number of molecules of mTRECs/100,000 thymocytes. We found that the absolute number of thymocytes dramatically decreased with age (P<0.05) and that molecules of mTREC/100,000 thymocytes also declined with mouse age (P<0.05). Splenocytes were isolated from aging mice and the frequency of naive phenotype CD4 and CD8 cells determined. There was a significant drop in both CD4 and CD8 naive peripheral T cells in the aged mice over time. mTREC analysis in purified CD4(+) and CD8(+) splenocytes demonstrated a constant level of mTRECs in the CD4 compartment until age 90 weeks, while the mTRECs in the CD8 compartment fell with age (P<0.05). By combining the mouse TREC assay with T cell phenotypic analysis, we demonstrated that IL-7 administration to young mice induced both increased thymopoiesis and peripheral T cell proliferation. In contrast, IL-7 treatment of aged mice did not augment thymopoiesis, nor induce expansion of splenic T cells. Thus, thymus output continues throughout murine adult life, and the thymic atrophy of aging in mice is not reversed by administration of IL-7. PMID- 11922944 TI - B cell activation leads to upregulated expression of the murine Sik-similar protein gene. AB - B cells stimulated by the combination of CD40L plus anti-Ig were screened for upregulated gene expression in an unbiased fashion through differential display. An inducible transcript was obtained that corresponds to a sequence previously isolated from pre-B cells and termed murine Sik-similar protein (mSSP). The mSSP gene is predominantly expressed in lymphoid tissues, including spleen and thymus, as well as testis, with lesser amounts in kidney and ovary. Among lymphocyte cell lines, mSSP expression varies widely. The mSSP protein is localized to the nucleus in NIH3T3 cells, and its expression in BAL-17 B cells varies with cell cycle progression. Expression of mSSP increased significantly within the first hours of B cell treatment with either CD40L, anti-IgM, or phorbol myristate acetate (PMA) with or without ionomycin. The predicted amino acid sequence of mSSP bears some homology to proteins involved in ribosomal RNA synthesis and processing. mSSP is a previously identified pre-B cell gene now shown for the first time to be an activation-responsive transcript in mature, primary B cells. PMID- 11922943 TI - c-Rel is a selective activator of a novel IL-4/CD40 responsive element in the human Ig gamma4 germline promoter. AB - Induction of isotype switching to a specific C(H) gene correlates with the transcriptional activation of the same gene in germline (GL) configuration. Expression of correctly spliced GL transcripts is necessary to target a switch region for recombination. In human B cells, the IgE and IgG4 isotypes are both induced by IL-4 through sequential switching, but are functionally antagonistic because IgG4 appears to have IgE-blocking activity. In order to understand the molecular mechanisms that regulate IgG4 production, we undertook a systematic analysis of the gamma4 GL promoter. A HindIII/NaeI region (-421/+474) highly conserved in the human gamma locus mediated the synergistic activation of a reporter gene in response to IL-4 and CD40 cross-linking. STAT6 binding to the proximal gamma4 GL promoter was essential for both IL-4-induced activation and CD40-dependent enhancement of transcription. Of note, a 45bp region (-76/-32) centered around the STAT6 binding motif drove robust synergistic activation of a heterologous fos promoter upon stimulation with IL-4 and CD40 cross-linking. This finding suggested that the (-76/-32) region may contain a novel IL-4/CD40 responsive element (RE). Electrophoretic mobility shift assays (EMSA) analysis using BL-2 nuclear extracts and in vitro translated NF-kappaB/Rel family proteins revealed the presence of a motif that overlaps the 5' end of the STAT6 element and binds selectively c-Rel. A mutation that abrogated c-Rel, but not STAT6, binding strongly impaired the CD40-induced enhancement of IL-4-dependent gamma4 GL transcription in reporter assays. These results indicate that c-Rel is selectively involved in the CD40-dependent activation of the IL-4/CD40 RE in the proximal gamma4 GL promoter. PMID- 11922945 TI - Molecular basis for the cross-reactivity of antibodies elicited by a natural anatoxin with alpha- and beta-toxins from the venom of Tityus serrulatus scorpion. AB - A non-toxic protein (TsNTxP) isolated from the venom of the noxious scorpion Tityus serrulatus (Ts) induces polyclonal antibodies cross-reactive with several toxins from the venom, in sharp contrast to anti-toxin antibodies which are toxin specific. To try to uncover the molecular basis for these unusual properties, peptide scanning experiments were performed and indicated that the N- and C terminal parts of TsNTxP enclose continuous epitopes (residues 1-15 and 47-61). Antibodies raised against peptides corresponding to these two regions were found to have neutralizing properties against a mixture of all toxic proteins from the T. serrulatus venom, indicating that residues 1-15 and 47-61 correspond to neutralizing epitopes. The identification of key antigenic residues within these two epitopes revealed that several of them are well conserved in the amino-acid sequences of the three main toxins (Ts II, Ts IV and Ts VII) from the venom: Glu 3, Tyr 5, Asp 8, Asp 50, Trp 55 and Lys 61. A single key-residue (Glu 58) is unique to TsNTxP. By using homology modeling, a model of the three-dimensional structure of TsNTxP was obtained. The antigenically important residues from TsNTxP were found to be surface exposed, with five of them clustered on the facet of the protein reported to enclose the active site of toxins. Residues equivalent to the seven key-residues of the anatoxin were also found to be exposed in the active toxins from T. serrulatus venom. These results show that antibodies elicited by the non-toxic protein TsNTxP recognized, within the N- and C-terminal parts of toxins of T. serrulatus, conserved and surface exposed residues which might also be involved in the toxic action of the proteins. PMID- 11922947 TI - Prediction of 'drug-likeness'. AB - Recent developments in combinatorial chemistry and high-throughput screening have dramatically increased the scale on which drug discovery programs are carried out. Along with these advances has come a need for automated methods of determining which compounds from a library should be synthesized and screened. These methods range from simple counting schemes to sophisticated machine learning techniques such as neural networks. While many of these methods have performed well in validation studies, the field is still in its formative stage. This paper reviews a number of computational techniques for identifying drug-like molecules and examines challenges facing the field. PMID- 11922948 TI - Prediction of intestinal permeability. AB - This review focuses on computational methods for the prediction of passive intestinal permeability. Existing computational models are surveyed and assessed in terms of descriptors, model type/complexity, speed of computation, predictive performance, and interpretability. Challenges to the successful computational prediction of intestinal permeability, i.e. data quantity, measurement imprecision, confounding factors such as solubility, metabolism, or active efflux, and the need for robust statistical methods, are also discussed. PMID- 11922949 TI - Computational approaches to the prediction of the blood-brain distribution. AB - This review attempts to summarise present knowledge related to the theoretical modelling of drug transport across the blood-brain barrier. Several computational protocols are described ranging from quantum mechanics-based approaches through molecular mechanics-related techniques to simple and fast procedures based on only the 2-D graph of the investigated structures. Amazingly, few descriptors have been shown to influence the derived relationships in a significant manner and a cornerstone in most of the described models are terms describing hydrogen bonding. A very quick quantitative assessment of the brain partitioning of a compound has also been devised using the following two rules: If N+O (the number of nitrogen and oxygen atoms) in a molecule is less than or equal to five, it has a high chance of entering the brain. The second rule predicts that if log P-(N+O) is positive then log BB is positive. PMID- 11922950 TI - Progress in understanding the structure-activity relationships of P-glycoprotein. AB - Efflux out of cells by P-glycoprotein (P-gp) represents a serious liability for pharmaceuticals, particularly for anti-cancer drugs. Consequently, identification of compounds as potential substrates is important for understanding their bioavailability. Also, the development of agents which reverse this multi-drug resistance phenotype has received considerable attention. Assays for determining these activities are reviewed. Recent literature and studies into the structure activity relationships (SAR) of the resulting data are discussed. Multiple binding sites and other complicating factors have prevented the development of a truly general, conclusive SAR either for substrate or inhibitory activities. Consequently, many models have tended to address only very general properties, such as lipophilicity and size. However, progress has been made in the last few years toward more specific SAR suggesting well-defined structural features responsible for both activities. The future of understanding the details of P-gp SAR lies in more specific assays that target specific binding sites and mechanisms of action. PMID- 11922951 TI - Modeling of active transport systems. AB - Transport proteins have critical physiological roles in nutrient transport and may be utilized as a mechanism to increase drug absorption. However, we have little understanding of these proteins at the molecular level due to the absence of high-resolution crystal structures. Numerous efforts have been made to characterize the P-glycoprotein efflux pump, the peptide transporter (PepT1) and the apical sodium-dependent transporter (ASBT) which are important not only for their native transporter function but also as drug targets to increase absorption and bioactivity. In vitro and computational approaches have been applied to gain some insight into these transporters with some success. This represents an opportunity for optimizing molecules as substrates for the solute transporters and providing a further screening system for drug discovery. Clearly the future growth in knowledge of transporter function will be led by integrated in vitro and in silico approaches. PMID- 11922952 TI - Prediction of drug solubility from structure. AB - The aqueous solubility of a drug is an important factor affecting its bioavailability. Numerous computational methods have been developed for the prediction of aqueous solubility from a compound's structure. A review is provided of the methodology and quality of results for the most useful procedures including the model implemented in the QikProp program. Viable methods now exist for predictions with less than 1 log unit uncertainty, which is adequate for prescreening synthetic candidates or design of combinatorial libraries. Further progress with predictive methods would require an experimental database of highly accurate solubilities for a large, diverse collection of drug-like molecules. PMID- 11922953 TI - Pharmacophore modeling of cytochromes P450. AB - Understanding the binding of ligands in the active site of a membrane-bound protein is difficult in the absence of a crystal structure. When these proteins are the enzymes involved in drug metabolism, it leaves little option but to use site-directed mutagenesis and in vitro studies to provide critical information relating to determinants of binding affinity. Pharmacophore models and three dimensional quantitative structure-activity relationships have been used either alone or in combination with protein homology models to provide this information for cytochrome P450s. At present, their application has been directed to the major enzymes but this may escalate in future as more in vitro data are generated for other P450s. The following review outlines the methodologies and models as well as future prospects for applying these technologies to P450s in the hope that future drugs will be selected with increased metabolic stability and fewer incidences of undesirable drug-drug interactions. PMID- 11922954 TI - Comparative modelling of cytochromes P450. AB - The superfamily of enzymes known as the cytochromes P450 (P450s) comprises a wide ranging class of proteins with diverse functions. They are known, amongst other things, to be involved in the hormonal regulation of metabolism and reproduction, as well as having a major clinical significance through their association with diseases such as cancer, diabetes and hepatitis. Knowledge of the three dimensional (3D) structure of a protein gives insight into its function. The 3D structures of P450s are therefore of considerable scientific interest. A number of high-resolution structures of P450s have been determined by X-ray crystallography and studies of these structures have provided valuable insights into the mechanism of these enzymes. Only one of these structures is mammalian and as yet there is no structural information on human P450s in the public domain. Until such a structure is solved it is necessary to employ alternative methods to gain structural insight into how human P450s perform their biological function. Here we report on the use of comparative modelling to predict the structure of human P450s based on knowledge of their amino acid sequences plus the 3D structures of other (not human) P450s. As an illustrative example of these techniques we have modelled the structure of P450 2C5 using five bacterial P450 structures as templates. We examine the importance of selecting suitable templates, obtaining a good amino acid sequence alignment, and evaluating the models generated. To improve the quality of the models an iterative cycle of sequence alignment, model building, and model evaluation is employed. The result is a model with excellent stereochemistry, good amino acid side chain environment properties, and a Calpha trace similar to the crystal structure. PMID- 11922955 TI - Computer systems for the prediction of xenobiotic metabolism. AB - The aim of pharmaceutical research and development is to ensure a continuing pipeline of new chemical entities (NCEs) displaying high therapeutic efficacy with few or no side effects. Failure of promising lead candidates late in the drug discovery processes is regarded as commercially unacceptable in today's increasingly competitive business environment. An inappropriate ADME/Toxicity profile in humans is the major cause of failure of lead candidates in late clinical stages of drug development. Combinatorial chemistry techniques coupled with high throughput screening protocols means that pharmaceutical companies are now dealing with an unprecedented number of NCEs on an annual basis. As a consequence, screening for undesirable ADME/Toxicity properties in the early stages of drug development, preferably pre-synthesis, is now considered the essential paradigm. In silico assessment of NCEs is rapidly emerging as the next wave of technology for early ADME/Toxicity prediction. In this review, we discuss the major commercially available products for the assessing the potential metabolic activity of xenobiotic substances in mammalian systems. PMID- 11922956 TI - Computer systems for the prediction of toxicity: an update. AB - In order to survive in the current economic climate, the pharmaceutical, agrochemical and personal product companies are required to produce large numbers of new, effective products whilst significantly reducing development time and costs. With the advent of combinatorial chemistry and high-throughput screening (HTS), the numbers of new candidate structures coming out of the discovery cycle has increased significantly. This has created a demand for faster screening of the toxicological properties of these candidates. Not surprisingly, computer methods for toxicity prediction offer an attractive solution to this problem because of their ability to screen large numbers of structures even before synthesis has occurred. In this paper the major, commercially available computer software systems for toxicity prediction are discussed together with their main strengths and limitations. PMID- 11922957 TI - Physiologically-based pharmacokinetic simulation modelling. AB - Drug selection is now widely viewed as an important and relatively new, yet largely unsolved, bottleneck in the drug discovery and development process. In order to achieve an efficient selection process, high quality, rapid, predictive and correlative ADME models are required in order for them to be confidently used to support critical financial decisions. Systems that can be relied upon to accurately predict performance in humans have not existed, and decisions have been made using tools whose capabilities could not be verified until candidates went to clinical trial, leading to the high failure rates historically observed. However, with the sequencing of the human genome, advances in proteomics, the anticipation of the identification of a vastly greater number of potential targets for drug discovery, and the potential of pharmacogenomics to require individualized evaluation of drug kinetics as well as drug effects, there is an urgent need for rapid and accurately computed pharmacokinetic properties. PMID- 11922958 TI - Bisphosphonates and atherosclerosis. AB - Bisphosphonates are used for the treatment of bone resorption, hypercalcemia, osteoporosis and Paget's disease. Etidronate, pamidronate and clodronate also inhibit the development of experimental atherosclerosis without altering serum lipid profile. Bisphosphonates inhibit the arterial calcification, lipid accumulation and fibrosis. They accumulate extensively in arterial walls and suppress macrophages in atheromatous lesions. In macrophage cultures, bisphosphonates inhibit the cellular accumulation and degradation of atherogenic LDL-cholesterol and foam cell formation. Further, they inhibit various enzymes involved in cell signal transduction and cholesterol biosynthesis. Recently, etidronate has been shown to inhibit the thickening of carotid arterial wall even in man. PMID- 11922959 TI - Responses of isolated perfused uterine vascular beds of nonpregnant and pregnant rats to endogenous and exogenous nitric oxide. AB - The responses to endothelial vasodilators and exogenous nitric oxide (NO) were characterized in intact isolated uterine vascular beds of nonpregnant, midpregnant and late-pregnant rats perfused with Kreb's buffer (37 degrees C, 5% CO(2) in air, pH approximately 7.4) containing 2% dextran and indomethacin. Phenylephrine increased perfusion pressure in the vascular beds equally in all three groups. In the presence of phenylephrine, N(omega)-nitro-L-arginine methyl ester (L-NAME) significantly augmented perfusion pressure in the order: nonpregnant14 degrees /s), the monocular hOKN becomes asymmetrical with a temporo to nasal preferred direction. For visual stimulation, we presented random-dot-patterns projected by a planetarium inside a perimeter. We tested four rotation axes of the planetarium, yaw (0 degrees -180 degrees ), roll (90 degrees -270 degrees ), diagonal (45 degrees -225 degrees ) and anti-diagonal (135 degrees -315 degrees ). In every position, the visual stimulus turned in clockwise and counter-clockwise direction. In a subregion of the pretectum of nine fish, we recorded 47 direction-selective neurons. Analysis of tuning-curves and preferred direction vectors show that these neurons encode both horizontal (yaw) and vertical (roll) visual stimulus directions. These results suggest that the control of horizontal and vertical OKN might not segregate into different nuclei in fish. PMID- 11923005 TI - Gbx2 expression in the late embryonic chick dorsal thalamus. AB - The expression pattern of the transcription factor gene Gbx2 in the forebrain of chicken embryos (embryonic day 14) was mapped using digoxigenin-labeled riboprobes and compared with the expression of the transcription factors Pax6 and Nkx2.2. The topographic analysis of Gbx2 expression on coronal and sagittal sections discriminated the positions and boundaries of diverse neuronal nuclei belonging to the dorsal thalamus from neighboring territories (the epithalamus, ventral thalamus, pretectum, and the underlying basal plate). The differential expression of Gbx2 within the dorsal thalamus clearly corresponds with the existence of four primary subdivisions identified in a previous study from this laboratory [13]: the anteroventral region and dorsal, intermediate, and ventral tiers. The subhabenular region turned out not to express Gbx2; this possibly implies it needs to be distinguished as a fifth separate dorsal thalamus subdivision. PMID- 11923006 TI - Organization of the mouse dorsal thalamus based on topology, calretinin immnunostaining, and gene expression. AB - To better understand the organization and evolution of the dorsal thalamus, we have made a first approach to analyze the possible histogenetic compartments of the mammalian dorsal thalamus using mouse embryos. For that, we have analyzed the expression of the proneural gene Math4a and the protein calretinin. Our results suggest the existence of rostrodorsal, caudoventral, and ventral compartments in the embryonic dorsal thalamus of the mouse, which partly parallel the dorsoventral histogenetic tiers postulated in the dorsal thalamus of sauropsids. The rostrodorsal compartment of the mouse dorsal thalamus is characterized by expression of Math4a, and it appears to include sensory and motor thalamic nuclei projecting to the dorsal pallium (isocortex). This compartment appears equivalent to the lemnothalamus proposed by Butler in tetrapods based on hodological grounds. The caudoventral and ventral compartments of the mouse dorsal thalamus lack expression of Math4a in the mantle, but they are characterized by several populations of calretinin-immunorective neurons that show projections to the claustroamygdaloid region in the ventrolateral pallium. More studies will be needed to analyze if the compartments proposed in this study represent true histogenetic units, and to find homologous developmental fields in all vertebrates. PMID- 11923007 TI - A proposed homology between the reptilian dorsomedial thalamic nucleus and the mammalian paraventricular thalamic nucleus. AB - We have compared the reptilian dorsomedial thalamic nucleus with the mammalian paraventricular thalamic nucleus from a topographic, chemoarchitectonic, and hodological point of view. Both nuclei are localized to a similar position in the dorsalmost aspect of the dorsal thalamus (midline nuclei). They also are uniformly calretinin-immunoreactive, both their cells and neuropil are strongly immunostained for calretinin, and the dorsomedial nucleus presents a strong calbindin immunoreactivity as well. Finally, the reptilian and the mammalian nuclei share a set of afferent and efferent connections with a number of forebrain structures. On the whole, this set of data allows us to propose that the dorsomedial nucleus and the paraventricular thalamic nucleus are homologous. Both represent an important relay station in pathways connecting the hypothalamus with telencephalic areas involved in visceral and motivational aspects of behavior, such as nucleus accumbens and the central amygdala. PMID- 11923008 TI - Afferent and efferent projections of the dorsal anterior thalamic nuclei in the lizard Podarcis hispanica (Sauria, Lacertidae). AB - The aim of this study was to investigate the afferent and efferent connections of the anterior thalamic nuclei in the lizard Podarcis hispanica. To identify potential sources of sensory inputs and to determine the fine organization of the projections of these thalamic nuclei to the telencephalon, we injected the sensitive tracer biotinylated dextran amine (BDA) into different nuclei of the anterior dorsal thalamus. We also injected BDA into several telencephalic areas in order to corroborate the results of thalamic injections. Our results show that the anterior thalamic nuclei receive projections from multiple areas and nuclei distributed throughout most of the brain, from rhombencephalon to telencephalon, and project to several telencephalic areas. The nucleus dorsolateralis anterior receives somatic (visual, somatosensory, auditory) as well as visceral inputs, and it is thus an important gateway for the relay of multisensory information to the telencephalon. PMID- 11923010 TI - Comparison of thalamic populations in mammals and birds: expression of ErbB4 mRNA. AB - The expression of ErbB4 mRNA was examined in dorsal thalamic regions of chicks and rats. In rats, ErbB4 expression was observed in the habenular nuclei, the paraventricular nucleus, intermediodorsal nucleus, the central medial thalamic nucleus, the posterior nucleus, the parafascicular nucleus, the subparafascicular nucleus, the suprageniculate nucleus, the posterior limitans nucleus, the medial part of the medial geniculate nucleus, the peripeduncular nucleus, the posterior intralaminar nucleus, the lateral subparafascicular nucleus, the lateral posterior nucleus, and all ventral thalamic nuclei. In chicks, expression was observed in the subhabenular nucleus, the dorsomedialis posterior nucleus, the dorsointermedius posterior nucleus, the nucleus of the septomesencephalic tract, and areas surrounding the rotundus and ovoidalis nuclei, including the subrotundal and suprarotundal areas, and all ventral thalamic nuclei. Most cells within ovoidalis and rotundus were not labeled. The similar pattern of afferent and efferent projections originating from ErbB4-expressing regions of the mammalian and bird dorsal thalamus suggests that ErbB4 hybridizing cells may be derived from a single anlage that migrates into multiple thalamic regions. Most neurons in the rotundus and ovoidalis nuclei of chick may be homologous to unlabeled clusters of neurons intermingled with ErbB4-expressing cells of the mammalian posterior/intralaminar region. PMID- 11923009 TI - Thalamo-telencephalic connections: new insights on the cortical organization in reptiles. AB - Tracer injections into the dorsal tier of the lacertilian dorsal thalamus revealed an extensive innervation of the cerebral cortex. The medial cortex, the dorsomedial cortex, and the medial part of the dorsal cortex received a bilateral projection, whereas the lateral part of dorsal cortex and the dorsal part of the lateral cortex received only an ipsilateral thalamic projection. Thalamocortical fibers were found superficially in all cortical regions, but in the dorsal part of the lateral cortex, varicose axons within the cellular layer were also observed. The bilateral thalamocortical projection originates from a cell population located throughout the dorsolateral anterior nucleus, whereas the ipsilateral input originates mainly from a rostral neuronal subpopulation of the nucleus. This feature suggests that the dorsolateral anterior nucleus consists of various parts with different projections. The dorsal subdivision of the lateral cortex displayed hodological and topological (radial glia processes) features of a dorsal pallium derivative. After tracer injections into the dorsal cortex of lizards, we found long descending projections that reached the striatum, the diencephalic basal plate, and the mesencephalic tegmentum, which suggests that it may represent a sensorimotor cortex. PMID- 11923011 TI - The pallial amygdala of amniote vertebrates: evolution of the concept, evolution of the structure. AB - Embryological studies indicate that the amygdala includes pallial structures, namely the cortical amygdala (olfactory and vomeronasal) and the basolateral complex deep to it. In squamate reptiles, the cortical amygdala includes secondary olfactory (the ventral anterior amygdala) and vomeronasal centres (the nucleus sphericus). In birds, the situation is far less clear, due to the relative underdevelopment of the chemosensory systems. The basolateral amygdala of squamate reptiles includes two ventropallial structures: the posterior dorsal ventricular ridge and the lateral amygdala. Like their mammalian counterparts, these centres give rise to glutamatergic projections to the striatal (centromedial) amygdala and the ventromedial hypothalamus. Using the same criteria, the caudal neostriatum and the ventral intermediate archistriatum may represent the ventral pallial amygdala of birds. The basal nucleus of the mammalian amygdala is a lateropallial territory. In reptiles, the lateral pallium includes the dorsolateral amygdala, which, like the mammalian basal nucleus, projects bilaterally to the striatum/accumbens and receives distinct cholinergic and dopaminergic innervations. In the avian brain, the same embryological, hodological, and histochemical criteria are met by the area temporo-parieto occipitalis, the caudolateral neostriatum and the dorsal intermediate archistriatum. Therefore, the projections from these structures to the paleostriatum and the lobus paraolfactorius are amygdalostriatal, rather than corticostriatal connections. PMID- 11923012 TI - Understanding the basic circuitry of the cerebral hemispheres: the case of lizards and its implications in the evolution of the telencephalon. AB - The organization of the cerebral hemispheres of mammals is characterized by corticostriatal glutamatergic projections and striatopallidal GABAergic ones, plus the descending projections of the pallium and subpallium to extratelencephalic targets. The present review of the available neuroanatomical data on the forebrain of lizards suggests that the telencephalon of reptiles also follows this basic pattern of connectivity. In addition, we show that this basic circuitry includes a pallido-cortical projection, therefore forming a cortico striato-pallido-cortical circuit. The analysis of this circuitry for the medial, dorsal, lateral, and ventral pallial divisions in reptiles and mammals leads to the following conclusions: (1) The medial and dorsal cortices of lizards together appear to be equivalent to the medial pallium of mammals. (2) The projection from the lacertilian dorsal cortex to the striatum proper resembles the subiculo striatal projection of mammals, rather than the isocortical projection to the caudatus-putamen. (3) Most of the dorsal striatum of reptiles is engaged in the corticostriatal circuit corresponding to the ventral pallium (the anterior dorsal ventricular ridge), and therefore, it is not equivalent to the mammalian caudatus putamen, which is involved in the circuit of the dorsal pallium. (4) The main and accessory olfactory bulbs also follow this pattern of connections. PMID- 11923013 TI - Development and evolution of the collopallium in amniotes: a new hypothesis of field homology. AB - Embryological development is uniquely positioned to illuminate both hodology in adult brains and its inherited genetic bases. The lateral corner of the lateral ventricle in mammals is a particularly crucial region where cell migration patterns, transiently formed connections, axonal growth kinetics, and fasciculation patterns are complex and interactive. Based on hodology, the sauropsid anterior dorsal ventricular ridge (ADVR) has been proposed as the homologue on a one-to-one basis of the mammalian lateral neocortex (LNC), the basolateral amygdalar complex (BLA), or the claustrum-endopiriform nucleus (CE). Data on gene expression patterns during development have indicated ADVR homology with parts of the latter two structures rather than with LNC. Collothalamic nuclei (the set of dorsal thalamic nuclei that receive their predominant input from the midbrain roof) project to part of BLA and to LNC. Recent findings demonstrate a complex pattern of mutually overlapping but noncongruent gene expression territories and collothalamic projections, which suggests a new, collopallial field hypothesis that the ADVR is homologous as a field to all three structures LNC, BLA, and CE. This hypothesis accounts for current hodological and developmental data as well as for lack of a CE in monotremes and for an abnormal subcortical lamina of gray matter that results from a genetic abnormality in humans. PMID- 11923014 TI - An hypothesis on the early evolution of the development of the isocortex. AB - We propose an hypothesis on the evolutionary origin of the unique inside-out developmental gradient of the isocortex, in which deep layers originate before superficial layers. This contrasts with the development of the reptilian cortex, which originates in an outside-in gradient. In mice, a mutated protein, reelin, produces the reeler phenotype, whose cortex has an outside-in neurogenetic gradient like in reptiles. Reelin is normally located in the marginal layer of the developing cerebral cortex, and its normal function has been proposed to be a stop signal that prevents radially migrating cells from moving into the marginal zone. Additionally, mutations on the kinase Cdk5, or in its neuronal-specific activator p35, produce a deficit similar to reeler in that the neurogenetic gradient is outside-in. However, contrary to reeler, in which no cell-sparse layer I is observed, in these mice, a well-defined layer I exists, which suggests that migrating cells respond normally to reelin. Apparently, Cdk5/p35 participate in permitting cortical cells to move across pre-existing (earlier produced) cortical layers, in order to be able to contact reelin once they reach the marginal zone. We suggest that the evolutionary advent of the mammalian cortical inside-out gradient became partly possible through the activation of the Cdk5/p35 pathway, which permitted migrating cells to move across layers of older cells. At about the same time, reelin became an important element in cortical development as it prevented neuronal migration into the marginal zone (cortical layer I) and facilitated the migration of neurons past postmigratory elements. PMID- 11923015 TI - Reelin immunoreactivity in the adult neocortex: a comparative study in rodents, carnivores, and non-human primates. AB - Recent evidence indicates that, in addition to playing a crucial role in early cortical development, intercellular signaling mediated by the protein Reelin may be widely active in the adult neocortex. The extent of Reelin distribution and its functional role in the adult are not clear yet. Here, we have examined Reelin immunoreactivity in the neocortex of an adult rodent (rat, Rattus norvegicus), a carnivore (ferret, Mustela putorius), and a primate (macaque monkeys Macaca nemestrina, Macaca mulatta) at the optic microscope level. Our data show that the neocortex of all three species contains several morphologically distinct populations of interneurons whose perikaryon and proximal dendritic processes are heavily immunoreactive for Reelin. The laminar distribution of these cells is species-specific. In addition, discrete reelin-immunoreactive pericellular structures are present in virtually all neocortical neurons of macaques. PMID- 11923016 TI - Patch/matrix patterns of gray matter differentiation in the telencephalon of chicken and mouse. AB - The mammalian striatum, a subpallial area, consists of two compartments (patches/striosomes and matrix) that differ in their neuronal birth dates, connectivity, neurochemistry, and molecular make-up. For example, members of the cadherin family of adhesion molecules (cadherin-8 and OL-protocadherin) are differentially expressed by the striosomes and the striatal matrix. A patch/matrix type of organization also has recently been found in the ventral hyperstriatum and the neostriatum of the chicken pallium, where cell clusters of similar birthdates ("isochronic" clusters) are surrounded by a matrix of cells that are born at a different time. Immunostaining with antibodies against cadherins reveals a similar arrangement of cell clusters. In the avian neostriatum, cadherin-7-positive cell clusters ("islands") are surrounded by a matrix of cells that express R-cadherin. The islands coincide, at least in part, with the isochronic cell clusters, as shown by pulse-labeling with bromodeoxyuridine. Likewise, isochronic clusters of the hyperstriatum ventrale relate to patchy heterogeneities in the cadherin-7 immunoreactivity pattern. Cadherins are known to mediate the aggregation and sorting of cells during development in many organs. Their differential expression by isochronic cell populations in the mammal subpallium and avian pallium suggests a common morphogenetic mechanism that regulates the formation of the patch/matrix patterns in these regions. PMID- 11923017 TI - Neuroepithelial origin of the insular and endopiriform parts of the claustrum. AB - The lateral and ventral pallia have been proposed as the source of neurons for the insular and endopiriform claustra, respectively. However, this correlation is controversial. Here, we analysed this relationship by labelling radial glia in coronal slices of the telencephalon of paraformaldehyde-fixed rabbit embryos (E18 E28) and newborn rabbits with an anti-vimentin antibody or with the fluorescent dye DiI. The radial glia that crossed the claustrum was anchored to the neuroepithelium of the lateral ventricular angle (LVA) at all ages studied. The LVA was deep at E18, but it subsequently become shallower, because of the apposition of the portion of its walls proximal to the vertex of the LVA. At E18, the radial glia that crossed most of the insular claustrum extended from the lateral wall of the LVA (presumptive lateral pallium), and the radial glia that crossed either the most ventral part of the insular claustrum or the endopiriform claustrum proceeded from the medial wall of the LVA (presumptive ventral pallium). These results suggest that although the endopiriform claustrum originates from the ventral pallium, the insular claustrum originates from both the lateral and the ventral pallial portions. PMID- 11923018 TI - Spatial memory and hippocampal pallium through vertebrate evolution: insights from reptiles and teleost fish. AB - The forebrain of vertebrates shows great morphological variation and specialized adaptations. However, an increasing amount of neuroanatomical and functional data reveal that the evolution of the vertebrate forebrain could have been more conservative than previously realized. For example, the pallial region of the teleost telencephalon contains subdivisions presumably homologous with various pallial areas in amniotes, including possibly a homologue of the medial pallium or hippocampus. In mammals and birds, the hippocampus is critical for encoding complex spatial information to form map-like cognitive representations of the environment. Here, we present data showing that the pallial areas of reptiles and fish, previously proposed as homologous to the hippocampus of mammals and birds on an anatomical basis, are similarly involved in spatial memory and navigation by map-like or relational representations of the allocentric space. These data suggest that early in vertebrate evolution, the medial pallium of an ancestral fish group that gave rise to the extant vertebrates became specialized for processing and encoding complex spatial information, and that this functional trait has been retained through the evolution of each independent vertebrate lineage. PMID- 11923019 TI - Continuity and discontinuity of the radial scaffolding in the forebrain of a lizard embryo. AB - We have used ventricular injections of the fluorescent carbocyanine dye DiI (Molecular Probes, Eugene, OR, USA) to demonstrate the scaffolding of the radial ventricular cells in the embryonic forebrain of lizards. The results reveal changes of density, orientation, or thickness of the radial processes between adjacent regions. On the whole, they support the idea that the hypothalamus together with the telencephalic stalk and the telencephalic hemispheres make up the anterior region of the neural tube or secondary prosencephalon, as proposed by the prosomeric model. In contrast, the thalamic eminence seems part of the diencephalon. PMID- 11923020 TI - GFAP expression withdraws--a trend of glial evolution? AB - This study is a summary of investigations in the last decade with several collaborators on representatives of different vertebrate stocks. The results suggest that in the main vertebrate stocks (Agnathi, Chondrichthyes, Actinopterygii, Sarcopterygii-Amniotes), which had their parallel brain evolutions from the laminar brains to the elaborated ones, the astroglia also developed in parallel, and had a common trend of evolution. With growing brain complexity, free astrocytes arose and tended to predominate, and the spontaneous glial fibrillary acidic protein (GFAP)-expression regressed, in several areas. In the mammalian, avian, teleost, and batoid brains, therefore, large areas display a paucity, almost a lack of GFAP-immunoreactivity. The GFAP-expression in the GFAP-free areas seems to be inducible only in the presence of free astrocytes. PMID- 11923021 TI - Functional circuitry of the avian basal ganglia: implications for basal ganglia organization in stem amniotes. AB - Histochemical, pathway tracing, and neuropeptide/neurotransmitter localization studies in birds, reptiles and mammals during the 1970s and 80s clearly showed that the telencephalon in all amniotes consists of a prominent ventrally situated subpallial region termed the basal ganglia, and a large overlying region involved in higher order information processing termed the pallium or cortex. These studies also showed that the basal ganglia in all extant amniote groups possessed neurochemically and hodologically distinct striatal and pallidal territories. More recently, studies of the localization of genes controlling regional brain development have confirmed the homology of the basal ganglia among amniotes. In our ongoing studies, we have identified several aspects of the functional organization of the basal ganglia that birds also share with mammals. These include: (1) an extensive glutamatergic "cortico"-striatal input and distinctive, cell-type specific localization of glutamate receptor subtypes; (2) an extensive, presumptively glutamatergic intralaminar thalamic input to striatal neurons; (3) an extensive dopaminergic input from the midbrain targeting both substance P (SP) type and enkephalin (ENK) type striatal projection neurons, with SP-type striatal neurons seemingly richer in the D-1 type dopamine receptor; and (4) SP+ and ENK+ striatal outputs giving rise to functionally distinct so-called direct and indirect motor output pathways, with the direct pathway having a pallido-thalamo motor cortex loop and the indirect pathway relaying back to the direct circuit via the subthalamic nucleus. These findings suggest that the major aspects of the cellular organization and functional circuitry of the basal ganglia in stem amniotes were already as observed in living amniotes, as therefore presumably was its key role in movement control. Because the organization of the basal ganglia of anamniotes is clearly less elaborate than in amniotes, and because the basal ganglia and cortex in amniotes are clearly extensively interconnected structures, it seems likely that stem amniotes were characterized by a major step forward in the grade of telencephalic organization of both the basal ganglia and the pallium. PMID- 11923022 TI - Electrophysiological analysis of a songbird basal ganglia circuit essential for vocal plasticity. AB - The discrete, interconnected nuclei of the songbird brain, collectively termed the song system, underlie the learning and production of song. Two main forebrain pathways have been identified that contribute to song production, learning, and adult plasticity. A posterior "motor pathway" including nucleus HVc (used as the proper name), the robust nucleus of the archistriatum (RA) and descending projections to the brainstem, is essential for song production. An "anterior forebrain pathway," arising from HVc, passing through area X of the lobus parolfactorius, the medial portion of the dorsolateral nucleus of the anterior thalamus and the lateral magnocellular nucleus of the anterior neostriatum, and finally terminating in RA, is essential for song learning and adult plasticity. The fact that the lobus parolfactorius is thought to form a part of the avian striatum implies several predictions for the connections of area X and for the properties of its neurons. Here, we review the existing anatomical and electrophysiological data bearing on the nature of area X as a striatal structure. In general, the data strongly favor the notion that area X is striatal. One set of observations, however, is at odds with that idea, and we provide and partially test the hypothesis that area X also contains a pallidal component. We discuss further tests of this idea and implications for thinking of the song system as a basal ganglia loop similar to that described in mammals. PMID- 11923023 TI - A putative striato-dorsal thalamic pathway in lizards. AB - Striatal targets related to the dorsal thalamus were studied in reptiles. The lateral striatum projects to globus pallidus and to three cellular groups associated to the lateral forebrain bundle: the anterior entopeduncular nucleus, the suprapeduncular nucleus, and the ventromedial thalamic nucleus. The projection is heavier on the suprapeduncular nucleus, which in turn projects on nucleus rotundus in the dorsal thalamus. Nucleus rotundus is the origin of a prominent projection to the lateral striatum among other forebrain areas. The intermediomedial striatum projects also to globus pallidus and to the three cellular groups associated with the lateral forebrain bundle, but in this case, the projection is heavier on the ventromedial thalamic nucleus. The latter nucleus targets a number of nuclear aggregates in the ventral tier of the dorsal thalamus, which in turn project to the intermediomedial striatum. As in mammals, the striatum in reptiles may influence through these pathways its input from the dorsal thalamus. PMID- 11923024 TI - Striato-amygdaloid transition area lesions reduce the duration of tonic immobility in the lizard Podarcis hispanica. AB - Neuroanatomical data suggest that the lizard striato-amygdaloid transition area is homologous with the mammalian central amygdala. In order to investigate possible functional similarities, tonic immobility was induced in adult lizards and its duration recorded. Each lizard was then randomly assigned to one of three treatments: (1) bilateral striato-amygdaloid transition area lesions, (2) bilateral dorsal cortex lesions or (3) untreated controls. Three days after trial 1, each lizard was subjected to a second trial and the tonic immobility duration recorded. The mean tonic immobility duration in lizards with striato-amygdaloid transition area lesions was significantly shorter (80.5%; p < 0.0033) in trial 2 than in trial 1. There were no inter-trial differences within dorsal cortex lesioned lizards or untreated controls. There was a significant treatment effect on tonic immobility duration in trial 2 (p < 0.0001). The mean tonic immobility duration of lizards with striato-amygdaloid transition area lesions was significantly shorter than that of dorsal cortex-lesioned lizards (72.2%; p < 0.01) or untreated controls (78.2%; p < 0.01). There was no significant difference in mean tonic immobility duration between dorsal cortex-lesioned lizards and untreated controls. Tonic immobility is considered to be an anti predator behaviour that reflects the underlying state of fear. Therefore, the reduced tonic immobility duration in lizards with striato-amygdaloid transition area lesions reflects a reduction of fear. These results provide the first data to indicate a functional similarity between the lizard striato-amygdaloid transition area and the mammalian central amygdala. PMID- 11923025 TI - Neural substrates for processing chemosensory information in snakes. AB - Snakes interact with their chemical environment through their olfactory and vomeronasal systems. The present report summarizes advances on neural substrates for processing chemosensory information. First, the efferent and centrifugal afferent connections of the main and accessory olfactory bulbs were reinvestigated. Second, the afferent and efferent connections of the nucleus sphericus, the main target of the accessory olfactory bulb, were characterized. The nucleus sphericus gives rise to a very small projection to the hypothalamus, but it does project to other telencephalic structures where olfactory and vomeronasal information could converge. Third, the intra-amygdaloid circuitry and the amygdalo-hypothalamic projections were described. The medial amygdala, for instance, receives both vomeronasal and olfactory inputs and projects to the hypothalamus, namely, to the lateral posterior hypothalamic nucleus. Fourth, because the lateral posterior hypothalamic nucleus projects to the hypoglossal nucleus, the motor center controlling the tongue musculature, this projection could constitute a pathway for chemosensory information to influence tongue flicking behavior. In summary, vomeronasal information is mostly relayed to the hypothalamus not via the nucleus sphericus but through other telencephalic structures. Convergence of olfactory and vomeronasal information appears to occur at different levels in the telencephalon. A neural substrate for the chemosensory control of tongue-flicking behavior is provided. PMID- 11923026 TI - Serotonergic innervation of the telencephalon in the domestic chick. AB - The serotonergic system in the telencephalon of the domestic chick was investigated using an antibody specific to serotonin (5-HT). Most parts of the forebrain, such as the different subdivisions of the visual Wulst and the neostriatum, displayed a rather uniform, moderate to dense innervation of serotonergic (5-HT+) fibers. However, some highly distinct area-specific differences could be observed. Primary sensory areas such as the ectostriatum, layer L2 of field L, and the rostral part of the nucleus basalis displayed very few 5-HT+ fibers. In contrast, the dorsal part of the archistriatum intermedium, the nucleus taeniae, a medial part of the lobus parolfactorius and the dorsomedial part of the hippocampus displayed an extremely dense serotonergic innervation. In general, three different types of 5-HT+ axons could be distinguished. The most common was a fine, highly varicose type, whereas beaded axons, exhibiting larger varicosities, and a thick non-varicose type, exhibiting occasional swellings, were much sparser. In summary, these findings indicate that the serotonergic innervation of the avian telencephalon is extensive but site specific, and is organized in a highly similar way to that in mammals. The high accumulation of 5-HT+ fibers in the dorsal part of the archistriatum intermedium points to a prominent role for 5-HT in fear behavior. PMID- 11923027 TI - Species differences in the projections from the entorhinal cortex to the hippocampus. AB - Both differences and similarities exist between mammalian species in the projections from entorhinal cortex to the hippocampal formation. In most species, layer II cells of the entorhinal cortex project to the dentate gyrus, and they terminate in the outer two-thirds of the molecular layer of the dentate gyrus. The axons from layer III cells project bilaterally to areas CA(1) and CA(3) of the hippocampus, terminating in the stratum lacunosum moleculare. We have analyzed these projections in mice, and in general, the entorhinal cortex-to hippocampus projections are similar to those in rats. Axons from layer II neurons terminate in the outer and middle thirds of the molecular layer of the dentate gyrus, and axons from layer III neurons terminate bilaterally in the stratum lacunosum moleculare of areas CA(1) and CA(3), and in the molecular layer of the subiculum. However, in contrast to rat, mouse entorhinal cortex neurons do not appreciably project to the contralateral dentate gyrus. Most species, including mice, show a similar topographical organization of the entorhinal-hippocampal projections, with neurons in the lateral part of both the lateral and medial entorhinal cortex projecting to the dorsal part or septal pole of the hippocampus, whereas the projection to the ventral hippocampus originates primarily from neurons in medial parts of the entorhinal cortex. PMID- 11923028 TI - Comparative aspects of the olfactory portion of the entorhinal cortex and its projection to the hippocampus in rodents, nonhuman primates, and the human brain. AB - The entorhinal cortex is a component of the hippocampal formation characterized by its projection to the dentate gyrus. Cortical input is relayed to the hippocampus through the entorhinal cortex. The entorhinal cortex can be recognized best in mammals, and it reaches its maximal importance in both human and nonhuman primates. In primates, but not in rodents, the olfactory projection from the olfactory bulb is the only direct sensory projection reaching the entorhinal cortex, where it defines a rostromedial subfield (olfactory subfield of the entorhinal cortex, EO). In this communication, we consider some comparative aspects of this projection in rodents, nonhuman primates, and human brains, in relation to the forward projection to the hippocampus. We base our analysis on the following three considerations: (1) Topology: Topologically, the entorhinal cortex lies medial to the rhinal fissure, well developed in all mammals except in humans. EO maintains the same topological relationship to the rhinal fissure in the three mammalian orders. (2) Cytoarchitecture: The rostromedial portion shows poorer lamination compared with caudomedial portions of the entorhinal cortex in the three mammalian orders examined. (3) Connectivity: In rodents, the olfactory projection covers the whole extent of the entorhinal cortex, whereas it decreases substantially in macaques, where it roughly covers 15% of the entorhinal cortex. The human EO as defined by topological and cytoarchitectonic criteria may be even less than in macaques. Interestingly, good evidence exists that all of these mammalian orders maintain the same topographical scheme of projection to the rostral end of the hippocampus. PMID- 11923029 TI - Functional asymmetry of emotions in primates: new findings in chimpanzees. AB - In the past 15 years, there have been a number of studies conducted on asymmetries in the perception and production of facial expressions in human and non-human primates as a means of inferring hemispheric specialization for emotions. We review these studies to assess continuity and discontinuity between species in these emotional processes. We further present new data on asymmetries in the production of facial expressions in a sample of captive chimpanzees. Objective measures (hemimouth length and area) and subjective measures (human judgement's of chimeric stimuli) indicate that chimpanzees' facial expressions are asymmetric, with a greater involvement of the left side of the face (right hemisphere) in the production of emotional responses. Left hemimouth was bigger than the right in the facial expressions of pant-hooting, play, and silent bared teeth (p < 0.05) and it extended laterally more than the right in the categories of pant-hooting, silent bared-teeth, and scream face (p < 0.05). Human judges also reported that the left side of the faces was emotionally more intense in the case of the play and silent bared-teeth categories (p < 0.01). Thus, chimpanzees, like humans and some other non-human primates, show a right hemisphere specialization for facial expression of emotions, which suggests that this functional asymmetry is homologous in all these species. PMID- 11923030 TI - Benefits, adverse effects and drug interactions of herbal therapies with cardiovascular effects. AB - Because the use of herbal therapies in the U.S. is escalating, it is essential to be aware of clinical and adverse effects, doses and potential drug-herb interactions. A consumer poll in 1998 indicated that one-third of respondents use botanical remedies, and nearly one in five taking prescription medications also used herbs, high-dose dietary supplements or both. An estimated 15 million adults are at risk for potential adverse interactions involving prescription medications and herbs or vitamin supplements, yet most practicing physicians have little knowledge of herbal remedies or their effects. Herbal products are marketed without the proof of efficacy and safety that the Food and Drug Administration (FDA) requires of drugs. The Dietary Supplement and Health Education Act of 1994 allocates responsibility to manufacturers for ensuring safety and efficacy with no specific requirements to submit documentation. Manufacturers may state a product's physiologic effects but may not make claims for the treatment or cure of specific diseases. Consumers and practitioners have little information about product safety, contraindications, interactions or effectiveness and are reliant on manufacturers to provide accurate labeling. Recently, the growing number of foods with herbs has raised concerns at the FDA, which requires evidence that food additives are safe. Considering that the growing appeal of herbal remedies is likely to continue, physicians, particularly cardiologists, must become familiar with the available cardiovascular information on herbs. This review highlights the existing data on the efficacy, adverse effects and interactions for herbal therapies that impact on the cardiovascular system. PMID- 11923031 TI - A contemporary overview of percutaneous coronary interventions. The American College of Cardiology-National Cardiovascular Data Registry (ACC-NCDR). AB - The American College of Cardiology (ACC) established the National Cardiovascular Data Registry (ACC-NCDR) to provide a uniform and comprehensive database for analysis of cardiovascular procedures across the country. The initial focus has been the high-volume, high-profile procedures of diagnostic cardiac catheterization and percutaneous coronary intervention (PCI). Several large-scale multicenter efforts have evaluated diagnostic catheterization and PCI, but these have been limited by lack of standard definitions and relatively nonuniform data collection and reporting methods. Both clinical and procedural data, and adverse events occurring up to hospital discharge, were collected and reported according to uniform guidelines using a standard set of 143 data elements. Datasets were transmitted quarterly to a central facility for quality-control screening, storage and analysis. This report is based on PCI data collected from January 1, 1998, through September 30, 2000.A total of 139 hospitals submitted data on 146,907 PCI procedures. Of these, 32% (46,615 procedures) were excluded because data did not pass quality-control screening. The remaining 100,292 procedures (68%) were included in the analysis set. Average age was 64 +/- 12 years; 34% were women, 26% had diabetes mellitus, 29% had histories of prior myocardial infarction (MI), 32% had prior PCI and 19% had prior coronary bypass surgery. In 10% the indication for PCI was acute MI < or =6 h from onset, while in 52% it was class II to IV or unstable angina. Only 5% of procedures did not have a class I indication by ACC criteria, but this varied by hospital from a low of 0 to a high of 38%. A coronary stent was placed in 77% of procedures, but this varied by hospital from a low of 0 to a high of 97%. The frequencies of in-hospital Q-wave MI, coronary artery bypass graft surgery and death were 0.4%, 1.9% and 1.4%, respectively. Mortality varied by hospital from a low of 0 to a high of 4.2%. This report presents the first data collected and analyzed by the ACC-NCDR. It portrays a contemporary overview of coronary interventional practices and outcomes, using uniform data collection and reporting standards. These data reconfirm overall acceptable results that are consistent with other reported data, but also confirm large variations between individual institutions. PMID- 11923032 TI - Development of a risk adjustment mortality model using the American College of Cardiology-National Cardiovascular Data Registry (ACC-NCDR) experience: 1998 2000. AB - OBJECTIVES: We sought to develop and evaluate a risk adjustment model for in hospital mortality following percutaneous coronary intervention (PCI) procedures using data from a large, multi-center registry. BACKGROUND: The 1998-2000 American College of Cardiology-National Cardiovascular Data Registry (ACC-NCDR) dataset was used to overcome limitations of prior risk-adjustment analyses. METHODS: Data on 100,253 PCI procedures collected at the ACC-NCDR between January 1, 1998, and September 30, 2000, were analyzed. A training set/test set approach was used. Separate models were developed for presentation with and without acute myocardial infarction (MI) within 24 h. RESULTS: Factors associated with increased risk of PCI mortality (with odds ratios in parentheses) included cardiogenic shock (8.49), increasing age (2.61 to 11.25), salvage (13.38) urgent (1.78) or emergent PCI (5.75), pre-procedure intra-aortic balloon pump insertion (1.68), decreasing left ventricular ejection fraction (0.87 to 3.93), presentation with acute MI (1.31), diabetes (1.41), renal failure (3.04), chronic lung disease (1.33); treatment approaches including thrombolytic therapy (1.39) and non-stent devices (1.64); and lesion characteristics including left main (2.04), proximal left anterior descending disease (1.97) and Society for Cardiac Angiography and Interventions lesion classification (1.64 to 2.11). Overall, excellent discrimination was achieved (C-index = 0.89) and application of the model to high-risk patient groups demonstrated C-indexes exceeding 0.80. Patient factors were more predictive in the MI model, while lesion and procedural factors were more predictive in the analysis of non-MI patients. CONCLUSIONS: A risk adjustment model for in-hospital mortality after PCI was successfully developed using a contemporary multi-center registry. This model is an important tool for valid comparison of in-hospital mortality after PCI. PMID- 11923033 TI - The impact of renal insufficiency on clinical outcomes in patients undergoing percutaneous coronary interventions. AB - OBJECTIVES: We sought to determine the effect of varying degrees of renal insufficiency on death and cardiac events during and after a percutaneous coronary intervention (PCI). BACKGROUND: Patients with end-stage renal disease have a high mortality from coronary artery disease. Little is known about the impact of mild and moderate renal insufficiency on clinical outcomes after PCI. METHODS: Cardiac mortality and all-cause mortality were determined for 5,327 patients undergoing PCI from January 1, 1994, to August 31, 1999, at the Mayo Clinic, based on the estimated creatinine clearance or whether the patient was on dialysis. RESULTS: In-hospital mortality was significantly associated with renal insufficiency (p = 0.001). Even after successful PCI, one-year mortality was 1.5% when the creatinine clearance was > or =70 ml/min (n = 2,558), 3.6% when it was 50 to 69 ml/min (n = 1,458), 7.8% when it was 30 to 49 ml/min (n = 828) and 18.3% when it was < 30 ml/min (n = 141). The 18.3% mortality rate for the group with < 30 ml/min creatinine clearance was similar to the 19.9% mortality rate in patients on dialysis (n = 46). The mortality risk was largely independent of all other factors. CONCLUSIONS: Renal insufficiency is a strong predictor of death and subsequent cardiac events in a dose-dependent fashion during and after PCI. Patients with renal insufficiency have more baseline cardiovascular risk factors, but renal insufficiency is associated with an increased risk of death and other adverse cardiovascular events, independent of all other measured variables. PMID- 11923034 TI - Limited role of coronary angioplasty and stenting in coronary spastic angina with organic stenosis. AB - OBJECTIVES: We investigated the efficacy of percutaneous coronary intervention (PCI) in patients with coronary spastic angina (CSA) and severe organic stenosis. BACKGROUND: Coronary spasm occurs at the site of organic stenosis in most patients with CSA and severe stenosis, whereas multivessel spasm occurs frequently in those with normal coronary arteries. The incidence of multivessel spasm and the efficacy of PCI in patients with CSA and severe stenosis have not been fully elucidated. METHODS: Forty-five patients with CSA and severe stenosis underwent spasm provocative testing with intracoronary acetylcholine before and 7 +/- 3 months after PCI (20 patients had angioplasty and 25 patients had stenting), when all patients were free of restenosis. RESULTS: Spasm was induced at the site of severe stenosis in 30 patients (66.7%) with (n = 12) or without (n = 18) spasm induced in another vessel. In the remaining 15 patients, spasm was induced at a different site in the stenotic vessel and/or in another vessel. Repeat provocative tests were performed in 43 of 45 patients. Although spasm was never induced at exactly the same site of the initial stenosis that had been dilated, spasm was induced at a different site in the dilated vessel and/or in another vessel, in 33 (76.7%) of 43 patients. Multivessel spasm occurred in 28 (62.2%) of 45 patients on one or both provocations. CONCLUSIONS: Spasm was frequently induced at a site different from the initial stenosis, even in the absence of restenosis after PCI. Calcium antagonists should be continued in most patients with CSA who show no restenosis after PCI. PMID- 11923035 TI - Transcutaneous detection of aortic arch atheromas by suprasternal harmonic imaging. AB - OBJECTIVES: The goal of the present study was to examine whether suprasternal harmonic imaging (SHI) (i.e., harmonic imaging from the suprasternal windows) can visualize protruding arch atheromas (PAAs) and reliably predict the presence or absence of significant lesions. BACKGROUND: Protruding arch atheromas are a major source of cerebral and peripheral embolism and probably the most frequent cause of stroke during cardiac catheterization and open-heart surgery. Preprocedural screening by transesophageal echocardiography (TEE) would be desirable but is limited by the nature of the examination. METHODS: Of 354 patients who underwent a TEE study in our laboratory during the study period, 106 were referred for detection of a source of embolism. Findings were classified based on the French Aortic Plaque study criteria as: 1) no or minimal atherosclerotic changes; 2) PAAs < 4 mm; 3) PAAs > or =4 mm or presence of a mobile component. RESULTS: Adequate transcutaneous image quality could be achieved in 89 patients (84%). Protruding arch atheromas were present in 42 patients (47%) and absent in 47 (53%). Positive and negative predictive values for large PAAs on TEE were 91% and 98%, respectively. In one case, SHI detected a complex PAA inaccessible for TEE due to interposition of the left bronchus as demonstrated by dual helical computed tomography. Inter-observer agreement for SHI was 91%. CONCLUSIONS: Suprasternal harmonic imaging reliably predicted or excluded the presence of PAAs in a sizable, consecutive group of patients referred to TEE for detection of a source of embolism. It represents an excellent screening test and provides complimentary views of regions, which may be blind spots for TEE. PMID- 11923036 TI - Soluble intercellular adhesion molecule-1 and long-term risk of acute coronary events in patients with chronic coronary heart disease. Data from the Bezafibrate Infarction Prevention (BIP) Study. AB - OBJECTIVES: The goal of this study was to assess soluble intercellular adhesion molecule-1 (sICAM-1) level as a predictor of future acute coronary events in patients with chronic coronary heart disease (CHD). BACKGROUND: Increased sICAM-1 concentration has been shown to be associated with the incidence of CHD in healthy persons. Its significance in patients with CHD has been scarcely investigated. METHODS: We designed a prospective, nested case-control study. Sera were collected from patients with CHD enrolled in a secondary prevention trial that evaluated the efficacy of bezafibrate in reducing coronary events. We measured baseline sICAM-1 concentration in the sera of patients who developed subsequent cardiovascular events (cases: n = 136) during follow-up (mean: 6.2 years) and in age- and gender-matched controls (without events: n = 136). RESULTS: Baseline serum concentrations of sICAM-1 were significantly higher in cases versus controls (375 vs. 350 ng/ml; p < 0.05). Each 100 ng/ml increase in sICAM-1 concentration was associated with 1.27 (95% confidence interval [CI]: 1.00 to 1.63) higher relative odds of coronary events. Soluble ICAM-1 concentration in the highest quartile (>394 ng/ml) was associated with significantly higher odds of coronary events (compared with the lowest quartile), even after multivariate adjustment (2.31, 95% CI: 1.02 to 5.50). After adding fibrinogen and total white blood cell count to the multivariate model, the relative odds were 2.12 (95% CI: 0.88 to 5.35) and 2.70 (95% CI: 1.10 to 7.05), respectively. CONCLUSIONS: Elevated sICAM-1 concentration in CHD patients is associated with increased risk of future coronary events independent of other traditional risk factors. PMID- 11923037 TI - Evaluation of carotid artery and aortic intima-media thickness measurements for exclusion of significant coronary atherosclerosis in patients scheduled for heart valve surgery. AB - OBJECTIVES: We assessed the value of carotid intima-media thickness (CIMT) and thoracic aorta intima-media thickness (AoIMT) in ruling out significant coronary artery disease (CAD) in patients scheduled for heart valve surgery. BACKGROUND: Evaluation of CAD is needed in most patients undergoing heart valve surgery because of the high surgical morbidity in patients with significant CAD, raising the need for sensitive tests to exclude CAD. Coronary angiography is the reference standard, but this invasive procedure is not cost-effective, because more than two-thirds of these patients do not have significant CAD. METHODS: In a pilot study, CIMT and AoIMT cutoff values separating low- from high-risk groups were determined in 96 patients by using receiver-operating characteristic curves. Then, a prospective study was conducted in 152 patients to determine the statistical power of these cutoff values used alone or in combination. In both studies, carotid artery ultrasonography and transesophageal echocardiography were performed before coronary angiography and valve surgery. RESULTS: In the pilot study, CIMT < 0.55 mm and AoIMT < 3 mm were excellent predictors of the absence of CAD. In the prospective study, CIMT and AoIMT criteria were independent predictors of significant CAD in these patients, as assessed by logistic regression analysis. Carotid IMT criterion had 100% sensitivity and 100% negative predictive value. For the AoIMT criterion, sensitivity was 98%, and negative predictive value 99%. Combining the two criteria did not change sensitivity and negative predictive value but increased specificity to 78%. CONCLUSIONS: Measurements of CIMT and AoIMT may be useful in selecting patients who do not require coronary angiography before heart valve surgery. PMID- 11923039 TI - Myocardial viability testing and impact of revascularization on prognosis in patients with coronary artery disease and left ventricular dysfunction: a meta analysis. AB - OBJECTIVES: This study pools data from published series examining late survival with revascularization versus medical therapy after myocardial viability testing in patients with severe coronary artery disease (CAD) and left ventricular (LV) dysfunction. BACKGROUND: Previous observational studies have suggested survival benefit in such patients if they are revascularized when myocardial viability is detected on imaging tests. METHODS: A MEDLINE database search returned 24 viability studies reporting patient survival using thallium perfusion imaging, F 18 fluorodeoxyglucose metabolic imaging or dobutamine echocardiography. Annual death rates were extracted, pooled and analyzed with a random effects model. The risk-adjusted relationship between severity of LV dysfunction, presence of viability and survival benefit associated with revascularization was assessed by meta-regression. RESULTS: There were 3,088 patients (2,228 men), ejection fraction 32 +/- 8%, followed for 25 +/- 10 months. In patients with viability, revascularization was associated with 79.6% reduction in annual mortality (16% vs. 3.2%, chi-square = 147, p < 0.0001) compared with medical treatment. Patients without viability had intermediate mortality, trending to higher rates with revascularization versus medical therapy (7.7% vs. 6.2%, p = NS). Patients with viability showed a direct relationship between severity of LV dysfunction and magnitude of benefit with revascularization (p < 0.001). There was no measurable performance difference for predicting revascularization benefit between the three testing techniques. CONCLUSIONS: This meta-analysis demonstrates a strong association between myocardial viability on noninvasive testing and improved survival after revascularization in patients with chronic CAD and LV dysfunction. Absence of viability was associated with no significant difference in outcomes, irrespective of treatment strategy. PMID- 11923038 TI - Postprandial endothelial activation in healthy subjects and in type 2 diabetic patients: role of fat and carbohydrate meals. AB - OBJECTIVES: To compare the effect of a high-fat meal and a high-carbohydrate meal (pizza), with and without antioxidant vitamins, on endothelial activation in healthy subjects and in patients with type 2 diabetes mellitus. BACKGROUND: The postprandial state is becoming increasingly acknowledged to affect some early events of atherogenesis. METHODS: In a randomized, observer-blinded, crossover study, 20 newly diagnosed type 2 diabetic patients and 20 age- and gender-matched healthy subjects received two meals at one-week intervals: a high-fat meal (760 calories) and an isoenergetic high-carbohydrate meal (non-cheese pizza). In all subjects, the same meals were repeated immediately following ingestion of vitamin E, 800 IU, and ascorbic acid, 1,000 mg. RESULTS: In normal subjects, the high-fat meal increased the plasma levels of tumor necrosis factor-alpha (TNF-alpha), interleukin-6 (IL-6), intercellular adhesion molecule-1 (ICAM-1) and vascular cell adhesion molecule-1 (VCAM-1), which were prevented by vitamins. No change in these parameters occurred after pizza ingestion or pizza ingestion with vitamins. In diabetic patients, basal concentrations of glucose, cytokines and adhesion molecules were significantly higher than in nondiabetic controls. Both meals significantly increased cytokine and adhesion molecule levels, but the increase was more sustained following the high-fat meal. There was no significant change from baseline when vitamin supplementation accompanied each meal. There was a relationship between changes in serum triglycerides and changes in TNF-alpha (r = 0.39, p < 0.01), IL-6 (r = 0.28, p < 0.05) and VCAM-1 (r = 0.25, p < 0.05), and between changes in plasma glucose and changes in IL-6 (r = 0.36, p < 0.01) and ICAM-1 (r = 0.31, p < 0.02). CONCLUSIONS: An oxidative mechanism mediates endothelial activation induced by post-meal hyperlipidemia and hyperglycemia. PMID- 11923040 TI - Myocardial viability and prognosis in patients with ischemic left ventricular dysfunction. PMID- 11923041 TI - Cardiac resynchronization therapy restores optimal atrioventricular mechanical timing in heart failure patients with ventricular conduction delay. AB - We characterized the relationship between systolic ventricular function and left ventricular (LV) end-diastolic pressure (LVEDP) in patients with heart failure (HF) and baseline asynchrony during ventricular stimulation. The role of preload in the systolic performance improvement that can be obtained in HF patients with LV stimulation is uncertain.We measured the maximum rate of increase of LV pressure, LVEDP, aortic pulse pressure (PP) and the atrioventricular mechanical latency (AVL) between left atrial systole and LV pressure onset in 39 patients with HF. Two subgroups were identified: "responder" if PP improved, or "nonresponder."Maximum hemodynamic improvement occurred at an atrioventricular (AV) delay that did not decrease LVEDP. Left ventricular and biventricular (BV) stimulation increased systolic hemodynamics significantly, despite no significant increase in LVEDP. All parameters decreased when the LVEDP was decreased by shorter AV delay. Left ventricular and BV stimulation provided better hemodynamics than right ventricular (RV) stimulation. For the nonresponder subgroup, systolic hemodynamics only worsened during AV delay shortening. For the responder subgroup, optimum PP was achieved when AVL was near zero. Restoration of optimal left atrial-ventricular mechanical timing partly contributes to the hemodynamic improvements observed in this patient subgroup. However, preload alone cannot explain the differences seen between RV and BV stimulation and the contradictory PP decreases even at maximal preload in the nonresponder subgroup. These results may be explained by a site-dependent mechanism such as the degree of ventricular synchrony. Caution should be taken in these patients when optimizing AV delays using echocardiography techniques that focus on LV inflow. PMID- 11923042 TI - Deconditioning fails to explain peripheral skeletal muscle alterations in men with chronic heart failure. AB - It remains controversial whether the skeletal muscle alterations in chronic heart failure (CHF) are due to disease pathophysiology or result from chronic deconditioning. The purpose of this study was to compare the skeletal muscle of CHF patients to peak oxygen consumption (peak VO(2)) matched sedentary controls. It has been established that skeletal muscle abnormalities are related to the exercise intolerance observed in patients with CHF. We studied the skeletal muscle of sedentary controls and patients with CHF matched for age, gender and peak VO(2). Hypothesis testing for the effects of group (CHF vs. normal), gender, and the interaction group x gender were performed. For capillary density only gender (p = 0.002) and the interaction of group x gender (p = 0.007) were significantly different. For 3-hydroxyl coenzyme A (CoA) dehydrogenase only group effect (p = 0.004) was significantly different. Mean values for capillary density were 1.46 +/- 0.28 for CHF men versus 1.87 +/- 0.32 for sedentary control men, 1.40 +/- 0.32 for CHF women versus 1.15 +/- 0.35 for sedentary control women. The activities for 3-hydroxyl CoA dehydrogenase were 3.09 +/- 0.88 for CHF men versus 4.05 +/- 0.42 for sedentary control men, 2.93 +/- 0.72 for CHF women versus 3.51 +/- 0.78 for sedentary control women. This study suggests that women and men adapt to CHF differently: men develop peripheral skeletal muscle abnormalities that are not attributable to deconditioning; women do not develop the same pathologic responses in skeletal muscle when compared with normal women matched for aerobic capacity. PMID- 11923043 TI - Reduction of insulin-like growth factor-I expression in the skeletal muscle of noncachectic patients with chronic heart failure. AB - OBJECTIVES: We sought to assess the role of insulin-like growth factor-I (IGF-I) in muscle wasting in chronic heart failure (CHF), serum concentrations and local muscular IGF-I expression were determined in patients with severe CHF. BACKGROUND: Chronic heart failure is associated with progressive muscle atrophy, leading to cardiac cachexia. Skeletal muscle disuse and inflammatory activation with elevated cytokine levels have been implicated; however, the pathomechanism involved remains largely unknown. METHODS: Serum levels of IGF-I were measured by competitive solid phase immunoassay in 47 patients with severe CHF (left ventricular ejection fraction < or =30%) and 15 age-matched healthy subjects (HS). Insulin-like growth factor-I and IGF-I receptor expression were assessed in vastus lateralis biopsies by real-time PCR and Western blot analysis. RESULTS: Although serum IGF-I was not significantly different (175 +/- 10 ng/ml in CHF vs. 170 +/- 12 ng/ml in HS, p = NS), local muscle IGF-I mRNA expression was reduced by 52% in CHF (6.7 +/- 0.4 vs. 14.0 +/- 0.9 arbitrary units in HS, p < 0.001). This was accompanied by an increase in IGF-I receptor mRNA expression (86.8 +/- 5.4 in CHF vs. 23.1 +/- 1.8 arbitrary units in HS, p < 0.001). Local IGF-I expression was significantly correlated with muscle cross-sectional area (R = 0.75, p = 0.01). Chronic heart failure patients with a body mass index of < 25 kg/m(2) showed signs of peripheral growth hormone (GH) resistance, as indicated by elevated serum GH, and reduced IGF-I levels. CONCLUSIONS: In CHF patients, muscle IGF-I expression is considerably reduced in the presence of normal serum IGF-I levels, possibly contributing to early loss of muscle mass. These findings are consistent with a potential role of IGF-I for skeletal muscle atrophy in CHF. PMID- 11923044 TI - Antecedent hypertension and heart failure after myocardial infarction. AB - OBJECTIVES: We sought to assess the relationship of antecedent hypertension to neurohormones, ventricular remodeling and clinical heart failure (HF) after myocardial infarction (MI). BACKGROUND: Heart failure is a probable contributor to the increased mortality observed after MI in those with antecedent hypertension. Hence, neurohormonal activation, adverse ventricular remodeling and a higher incidence of clinical HF may be expected in this group. However, no previous report has documented serial postinfarction neurohumoral status, serial left ventricular imaging and clinical outcomes over prolonged follow-up in a broad spectrum of patients with and without antecedent hypertension. METHODS: Inpatient events were documented in 1,093 consecutive patients (436 hypertensive and 657 normotensive) with acute MI. In 68% (282 hypertensive, 465 normotensive) serial neurohormonal sampling and radionuclide ventriculography were performed one to four days and three to five months after infarction. Clinical outcomes were recorded over a mean follow-up of two years. RESULTS: Plasma neurohormones were significantly higher in hypertensives than in normotensives one to four days and three to five months after infarction. From similar initial values, left ventricular volumes increased significantly in hypertensives, compared with normotensives. Left ventricular ejection fraction rose significantly in normotensive but not hypertensive patients. Together with higher inpatient (8.1% vs. 4.4%, p < 0.002) and post-discharge mortality (9.5% vs. 5.5%, p = 0.043), hypertensive patients incurred more inpatient HF (33% vs. 24%, p < 0.001) and more late HF requiring readmission to hospital (12.4% vs. 5.5%, p < 0.001). Antecedent hypertension predicted late HF in patients >64 years of age with neurohormonal activation and early left ventricular dilation. CONCLUSIONS: Antecedent hypertension interacts with age, neurohumoral activation and early ventricular remodeling to confer greater risk of HF after MI. PMID- 11923045 TI - Severe energy deprivation of human hibernating myocardium as possible common pathomechanism of contractile dysfunction, structural degeneration and cell death. AB - OBJECTIVES: We tested the hypothesis that severe alterations in myocardial energy metabolism play an important role in the pathophysiology of human hibernating myocardium (HHM). BACKGROUND: It is well established that a disturbed myocardial energy metabolism results in impairments of contractile performance, structure and viability. All of these are important characteristics of HHM. METHODS: In 16 patients with documented coronary artery disease and impaired left ventricular function, HHM was preoperatively detected by thallium-201 scintigraphy, radionuclide ventriculography and low-dose dobutamine echocardiography. These regions were validated as HHM by their recovery of contractile function three months following revascularization. During open-heart surgery, transmural biopsies were removed from the hibernating areas and analyzed both biochemically and morphologically. These findings were compared to normal human myocardium. All metabolite contents given were normalized for the degree of fibrosis (control: 9.8 +/- 0.5%; HHM: 28.1 +/- 3.0%; p < 0.05), providing myocellular contents. RESULTS: In HHM, decreased contents (micromol/g wet weight) in adenosine triphosphate (ATP) (control: 4.17 +/- 0.26; HHM: 1.72 +/- 0.25; p < 0.001), creatine phosphate (5.67 +/- 0.70 vs. 0.84 +/- 0.13; p < 0.001) and creatine (27.6 +/- 3.19 vs. 11.2 +/- 1.56; p < 0.0001) were found, but contents in lactate (2.22 +/- 0.26 vs. 25.38 +/- 3.53; p < 0.001), purine bases (0.58 +/- 0.09 vs. 1.26 +/- 0.13; p < 0.001) and protons (pH units: 7.199 +/- 0.01 vs. 6.59 +/- 0.07; p < 0.001) were increased. Levels in adenosine diphosphate, adenosine monophosphate and inorganic phosphate remained unchanged. Energy depletion in HHM was reflected by decreases in the free energy of ATP hydrolysis and in energy charge. CONCLUSIONS: These data confirm our hypothesis that HHM is energy depleted myocardium, exhibiting signs of chronic reduction in resting blood flow and a downregulation of energy turnover. The alterations in energy metabolism observed may become operative in triggering and maintaining contractile dysfunction, continuous tissue degeneration and cardiomyocyte loss. PMID- 11923046 TI - The effect of supplemental L-arginine on tolerance development during continuous transdermal nitroglycerin therapy. AB - OBJECTIVES: This study was designed to assess the effect of oral L-arginine on the development of tolerance during continuous transdermal nitroglycerin (TD-GTN) therapy. BACKGROUND: Continuous TD-GTN therapy leads to complete tolerance within 24 to 48 h. The mechanism(s) responsible for nitrate tolerance are unclear, but there is increasing evidence that nitroglycerin (GTN) leads to superoxide anion production. The trigger for this is unknown, but there is evidence that GTN alters nitric oxide synthase (NOS) function and also leads to reduced L-arginine availability at its site of action with NOS. METHODS: Fourteen patients with stable angina pectoris and reproducible treadmill walking time (TWT) until the onset of moderate angina were studied in a placebo-controlled, crossover study. Transdermal GTN (0.4 mg/h) was applied daily for two periods of 5 to 10 days with the patch left in place for 24 h each day. Capsules containing L-arginine (700 mg) or placebo were administered four times daily during a period of TD-GTN therapy. Treadmill walking time was determined before and 4 h after study capsules on day 1 before TD-GTN to assess the effect of L-arginine on exercise performance. On the last day, TWT was determined at 0 h (24 h after TD-GTN and 9 h after study capsule) and 4 h after TD-GTN reapplication and study capsule. After a 5 to 10 day washout period, the study was repeated with the opposite study capsule. RESULTS: Treadmill walking time until the onset of moderate angina was not influenced by the short-term administration of L-arginine. During continuous TD-GTN, the administration of L-arginine increased TWT 4 h and 24 h after patch application. This was significantly greater than TWT during administration of placebo capsules (p < 0.05). CONCLUSIONS: The administration of L-arginine modified or prevented the development of nitrate tolerance during continuous TD-GTN therapy. PMID- 11923048 TI - Endocarditis beyond the annulus: eat your heart out. PMID- 11923047 TI - Periannular extension of infective endocarditis. AB - OBJECTIVES: This prospective study was designed to assess the current clinical course, risk factors, microbiologic profile and echocardiographic findings of patients with left-sided endocarditis and perivalvular complications. BACKGROUND: Periannular complications worsen the prognosis of patients with endocarditis. The relation between these complications and the clinical and microbiologic data has not been clearly defined. METHODS: In this clinical cohort study, 211 patients with left-sided endocarditis, according to the Duke criteria, were prospectively recruited. All patients underwent conventional and transesophageal echocardiography. The mean follow-up interval was 151 days. RESULTS: Perivalvular complications were detected in 78 patients (37%). The incidence of periannular extension of infection in native and prosthetic valves was 29% and 55%, respectively. The presence of prosthesis (relative risk [RR] 1.88, 95% confidence interval [CI] 1.35 to 2.64) and previous endocarditis (RR 1.78, 95% CI 1.16 to 2.7) were the only pre-existing heart conditions associated with perivalvular complications. Aortic infection (RR 1.8, 95% CI 1.23 to 2.66) and the development of atrioventricular (AV) block (RR 2.55, 95% CI 1.91 to 3.41) were related with the existence of these complications. Coagulase-negative staphylococci were very common in patients with perivalvular complications (RR 1.77, 95% CI 1.21 to 2.59), and small vegetations were more frequent in these patients (RR l.45, 95% CI 0.95 to 2.22). An operation was more frequently performed in patients with perivalvular complications, but mortality was similar in patients with and without these complications. CONCLUSIONS: Aortic infection, prosthetic endocarditis, new AV block and coagulase-negative staphylococci were independent risk factors of periannular complications. The period between symptom onset and diagnosis, the incidence of pericardial effusion and persistent signs of infection were similar between patients with and without perivalvular complications. Patients with perivalvular complications did not demonstrate a difference in the presence or size of vegetations or the frequency of embolism. An operation was more frequently performed in these patients, but mortality was similar in both groups. PMID- 11923049 TI - Echocardiographic predictors of adverse outcomes in primary pulmonary hypertension. AB - OBJECTIVES: The aim of this study was to evaluate the relationships between echocardiographic findings and clinical outcomes in patients with severe primary pulmonary hypertension (PPH). BACKGROUND: Primary pulmonary hypertension is associated with abnormalities of right heart structure and function that contribute to the poor prognosis of the disease. Echocardiographic abnormalities associated with PPH have been described, but the prognostic significance of these findings remains poorly characterized. METHODS: Echocardiographic studies, invasive hemodynamic measurements and 6-min walk tests were performed and outcomes prospectively followed in 81 patients with severe PPH. Subjects were participants in a 12-week randomized trial examining the effects of prostacyclin plus conventional therapy compared with conventional therapy alone. RESULTS: During the mean follow-up period of 36.9 +/- 15.4 months, 20 patients died and 21 patients underwent transplantation. Pericardial effusion (p = 0.003) and indexed right atrial area (p = 0.005) were predictors of mortality. Pericardial effusion (p = 0.017), indexed right atrial area (p = 0.012) and the degree of septal shift in diastole (p = 0.004) were predictors of a composite end point of death or transplantation. In multivariable analyses incorporating clinical, hemodynamic and echocardiographic variables, pericardial effusion and an enlarged right atrium remained predictors of adverse outcomes. Six-minute walk results, mixed venous oxygen saturation and initial treatment randomization were also independently associated with a poor prognosis. CONCLUSIONS: Pericardial effusion, right atrial enlargement and septal displacement are echocardiographic abnormalities that reflect the severity of right heart failure and predict adverse outcomes in patients with severe PPH. These characteristics may help identify patients appropriate for more intensive medical therapy or earlier transplantation. PMID- 11923050 TI - A comparison of mechanical and laser transmyocardial revascularization for induction of angiogenesis and arteriogenesis in chronically ischemic myocardium. AB - OBJECTIVES: The purpose of the present study was to compare the use of a mechanical transmyocardial implant (TMI) device with transmyocardial laser revascularization (TMR) for induction of therapeutic angiogenesis and arteriogenesis in the chronically ischemic heart. BACKGROUND: Prior experimental studies have demonstrated evidence for neovascularization after both mechanical and laser transmyocardial revascularization, although a long-term comparison of the two techniques has not been performed. METHODS: Using an established model of chronic hibernating myocardium, mini-swine underwent 90% proximal left circumflex (LCx) coronary artery stenosis. One month later, baseline positron emission tomography (PET) and dobutamine stress echocardiography (DSE) were performed to quantitate regional myocardial blood flow (MBF) and function. Animals then underwent TMR with a holmium:yttrium-aluminum-garnet (holmium:YAG) laser (n = 5), TMI (n = 5), or sham redo-thoracotomy (n = 5). In the TMR group, the entire LCx region was treated with transmural laser channels at a density of 1/cm(2). Transmyocardial implants were placed transmurally at a similar density in the LCx region of the TMI group. Six months later, the PET and DSE studies were repeated, and the animals were euthanized. RESULTS: Six months after TMR, there was a significant increase over baseline in resting MBF to the lased LCx region (68.9 +/- 4.6% vs. 89.3 +/- 3.0% reference non-ischemic septal segments; p < 0.001). This increased MBF was accompanied by a significant improvement in LCx regional wall motion during peak dobutamine stress (p = 0.04). Compared with baseline, there was no change in LCx region MBF six months after either TMI (72.9 +/- 4.8% vs. 85.7 +/- 3.4%; p = 0.10) or sham redo-thoracotomy (75.6 +/- 4.6% vs. 80.1 +/- 5.0%; p > 0.2). Likewise, there was no significant change in rest or stress wall motion by DSE six months postoperatively in either group. Overall vascular density was increased only in the TMR-treated regions six months postoperatively. The difference between groups was most notable for a twofold increase in the number of small arterioles seen in the lased (4.4 +/- 0.3 arterioles per high power field; p < 0.001 vs. both TMI and sham) compared with TMI (2.2 +/- 0.2) and sham (1.9 +/- 0.2)-treated regions. CONCLUSIONS: Mechanical transmyocardial revascularization with a TMI device does not appear to promote physiologically significant angiogenesis or arteriogenesis in the chronically ischemic porcine heart and cannot be recommended for clinical trials at this time. Infrared laser mediated injury mechanisms may be important for inducing therapeutic neovascularization with direct myocardial revascularization techniques. PMID- 11923051 TI - Improved myocardial ischemia/reperfusion injury in mice lacking tumor necrosis factor-alpha. AB - OBJECTIVES: This study sought to assess the role of tumor necrosis factor-alpha (TNF-alpha) in myocardial ischemia/reperfusion (I/R) injury using TNF-alpha knockout (KO) mice. BACKGROUND: Tumor necrosis factor-alpha is thought to be involved in the pathogenesis of myocardial I/R injury by promoting leukocyte infiltration of the myocardium. However, the precise role of TNF-alpha in I/R injury is still unknown. METHODS: The hearts in TNF-alpha KO and wild-type (WT) mice were exposed by left lateral thoracotomy, and the left coronary artery was occluded for 30 min then reperfused for 120 min. RESULTS: The infarct size in TNF alpha KO mice was significantly reduced compared with WT mice. The frequency of arrhythmia was decreased, and cardiac function during reperfusion was significantly improved in TNF-alpha KO mice compared with WT mice. The activation of nuclear factor-kappaB (NF-kappaB), the expression of chemokines and adhesion molecules and the infiltration of leukocytes were also significantly reduced in TNF-alpha KO mice, compared with WT mice. These findings provide evidence that TNF-alpha aggravates I/R injury. CONCLUSIONS: Tumor necrosis factor-alpha exacerbates myocardial I/R injury at an early stage of reperfusion by activating NF-kappaB, thereby inducing chemokines and adhesion molecules and facilitating leukocyte infiltration. PMID- 11923052 TI - Heart hospitals: are they the next wave? PMID- 11923053 TI - President's page: convocation address: the art of medicine. PMID- 11923054 TI - ACC revised recommendations for training in adult cardiovascular medicine. Core Cardiology Training II (COCATS 2). (Revision of the 1995 COCATS training statement). PMID- 11923055 TI - Factors confounding the successful extrapolation of in vitro CYP3A inhibition information to the in vivo condition. AB - For the most part, the majority of adverse drug-drug interactions, which are pharmacokinetic in origin, can be understood in terms of alterations of cytochrome P450-catalyzed reactions. Much is known about the human P450 enzymes, and in many cases it is possible to apply this information to clinically related issues. Of the relatively small subset of the total number of human P450s, CYP3A4 appears to be responsible for the largest fraction of the drug oxidation reactions. As a consequence many important drug-drug interactions observed in the clinic are associated with drugs which are principally metabolized by CYP3A4. The two major reasons for drug-drug interactions involving CYP3A4 are induction and inhibition, with inhibition appearing to be the more important in terms of known clinical problems. Fortunately, with the available knowledge of human P450s and in vitro reagents, it is possible to do experiments with drugs to predict the in vivo condition. The goal of these studies is not only to improve predictions about which drugs might show serious P450 interaction problems, but also to decrease the number of in vivo interaction studies that must be performed in drug development. The focus of the current report is to describe some of the confounding factors associated with in vitro drug inhibition studies and the impact of these issues on in vitro/in vivo extrapolations. PMID- 11923056 TI - Pharmaceutical education in the wake of genomic technologies for drug development and personalized medicine. AB - The development of safe and effective new therapeutics is a long, difficult, and expensive process. Over the last 20-30 years, recombinant DNA (rDNA) technology has provided a multiple of new methods, molecular targets and DNA-based diagnostics to pharmaceutical research that can be utilized in assays for screening and developing potential biopharmaceutical drugs. In parallel, new innovative approaches to drug delivery systems were discovered and reached the market. Pharmaceutical biotechnology, pharmacogenomics, combinatorial chemistry, in close relation to high-throughput screening technologies, and bioinformatics are major advances that give a new direction to pharmaceutical sciences. To meet with the needs of this new dynamic era of pharmaceutical research and health care environment, pharmaceutical education has to set new priorities to keep pace with the challenges related to genomic technologies. The development of new initiative education programs, for both undergraduate and graduate curricula, in pharmacy has to be focused on preparing pharmacists oriented for both pharmacy practice and drug research and development. This can be achieved by providing future pharmacists with knowledge, skills and attitudes to be more competitive in the health care system, pharmacy practice-related fields, pharmaceutical industry and drug research and development areas, or finally in academia. Educators and pharmacy school members have the responsibility of deciding how, to what extent, by which methods, and/or in which way these changes and new directions in the education programs should be developed. PMID- 11923057 TI - Pharmacokinetics and effects of succinylcholine in African elephant (Loxodonta africana) and impala (Aepyceros melampus). AB - The phenomenon of slow onset of succinylcholine (Sch) effect in elephants was investigated by analyzing blood concentrations of Sch and its metabolite choline in elephant and impala. To assess whether the slow onset phenomenon is related to the pharmacokinetics of Sch following i.m. administration, we analyzed the time course of plasma concentrations of intact drug and its metabolite and determined its pharmacological effects. Blood samples were obtained from anaesthetized elephant (n=6) and impala (n=7) following i.m. administration of a lethal dose of Sch. Time from Sch injection to onset of apnoea and to death was significantly longer for elephant than impala (mean+/-S.D. apnoea 4.4+/-1.5 and 2.3+/-0.9 min, respectively; death 32.6+/-7.3 and 6.2+/-3.4 min, respectively). The C(max) was not different between elephants and impala (20.3+/-7.9 vs. 14.4+/-6.8 nmol ml-1, respectively) but the t(max) was significantly longer for elephants (23.0+/-7.6 vs. 3.7+/-2.2 min). Analysis of the plasma Sch and choline concentrations over time revealed that the relative amount of Sch entering the circulation within the first 30 s after i.m. injection is greater for impala than elephant. No greater rate in the plasma hydrolysis of Sch in elephant compared to impala was apparent. PMID- 11923059 TI - The effect of amiloride on the in vivo effective permeability of amoxicillin in human jejunum: experience from a regional perfusion technique. AB - The purpose of this human intestinal perfusion study (in vivo) was twofold. Firstly, we aimed to determine the effective in vivo jejunal permeability (P(eff)) of amoxicillin and to classify it according to the Biopharmaceutics Classification System (BCS). Secondly, we investigated the acute effect of amiloride on the jejunal P(eff) of amoxicillin. Amoxicillin, a beta-lactam antibiotic, has been reported to be absorbed across the intestinal mucosa by both passive diffusion and active transport. A regional single-pass perfusion of the jejunum was performed using a Loc-I-Gut perfusion tube in 14 healthy volunteers. Each perfusion lasted for 200 min and was divided into two periods of 100 min each. The concentration of amoxicillin entering the jejunal segment was 300 mg/l in both periods, and amiloride, an inhibitor of the Na+/H+ exchanger, was added at 25 mg/l in period 2. The concentrations of amoxicillin and amiloride in the outlet jejunal perfusate were measured with two different HPLC-methods. Antipyrine and [14C]PEG 4000 were added as internal standards to the perfusion solution. Amiloride had no significant effect on the jejunal P(eff) of amoxicillin. The human in vivo jejunal P(eff) for amoxicillin was 0.34+/-0.11 x 10(-4) and 0.46+/-0.12 x 10(-4) cm/s in periods 1 and 2, respectively. The high jejunal P(eff) for amiloride was 1.63+/-0.51 x 10(-4) cm/s which predicts an intestinal absorption of more than 90%. Following the BCS amoxicillin was classified as a low P(eff) drug, and amiloride had no acute effect on the in vivo jejunal P(eff) of amoxicillin. PMID- 11923058 TI - A novel method of producing a microcrystalline beta-sitosterol suspension in oil. AB - This paper describes a novel method of producing a microcrystalline oral suspension containing beta-sitosterol in oil for the treatment of hypercholesterolaemia. beta-Sitosterol pseudopolymorphs with different water contents were crystallized from acetone and acetone-water solutions. Structural analyses of the crystals were performed by Karl-Fisher titration, thermogravimetric analyses, X-ray diffraction and near infrared spectroscopy. The suspensions studied were composed of different concentrations of beta-sitosterol, oil and water. Suspensions were prepared by crystallization of hot concentrated solution of beta-sitosterol in oil by cooling with simultaneous agitation and water addition. The structural analyses of the suspensions were performed by X ray diffraction, near infrared spectroscopy and optical microscopy. The viscosity of the suspensions was analysed as a function of shear stress. beta-Sitosterol was observed to exist in three different forms: anhydrous, hemihydrated and monohydrated crystals. By changing both the beta-sitosterol and the water concentration of the suspension, the crystal size and shape could be controlled. Addition of water resulted in the formation of monohydrated needle-shaped crystals instead of platy-like anhydrous crystals. Needle-shaped particles formed structured suspensions with shear thinning behaviour. By increasing the volume fraction of solid particles in suspension by increasing the water and/or sterol concentration, the viscosity increased. A high sterol concentration resulted in high supersaturation and thus formation of small crystals. PMID- 11923060 TI - Formulation-dependent food effects demonstrated for nifedipine modified-release preparations marketed in the European Union. AB - The objective of this study was a comparative investigation of the influence of concomitant food intake on the bioavailability of two nifedipine-containing controlled-release formulations. Adalat OROS and CORAL were compared in a randomised, non-blind, four-way crossover design in 24 healthy, male subjects after single dose administration following a high fat American breakfast or an overnight fast of 12 h, respectively. Plasma samples were withdrawn until 48 h post-dose. In the fasted state, the bioavailability (AUC and C(max) values) was lower for CORAL than for Adalat OROS. Under fed conditions, differences in bioavailability between both products were markedly increased. With respect to the therapeutic use of both products, the most important finding was the significant dose-dumping effect observed after fed administration of CORAL, resulting in nifedipine plasma concentrations of nearly three- to four-fold in 11 of 24 volunteers. The mean ratio of C(max) was 235% comparing CORAL with Adalat OROS under these conditions. The formulation-dependent food interaction observed in this study may be therapeutically relevant, especially in the case of changing administration conditions or switching from one product to the other. PMID- 11923061 TI - Characterization of the pharmacokinetics of buprenorphine and norbuprenorphine in rats after intravenous bolus administration of buprenorphine. AB - The purpose of this project was to characterize the dose-dependent pharmacokinetics of buprenorphine (BN) and norbuprenorphine (NBN), the primary metabolite, after intravenous administration of different doses of BN to rats. Adult male Sprague-Dawley rats were divided into six groups and received a single intravenous bolus dose of 0.1, 0.3, 1, 3, 10 or 30 mg/kg of BN. A separate study was performed where BN and NBN were simultaneously administered intravenously (1 mg/kg+1 mg/kg). Plasma samples were obtained by centrifugation of the blood and analyzed for BN and NBN using a sensitive and selective gas chromatographic-mass spectrometric (GC-MS) bio-analytical method. Noncompartmental and compartmental methods were used to perform pharmacokinetic data analysis. BN declined triexponentially with a dose-dependent increase in its volume of distribution, V(ss) (8.37-18.2 l/kg) and clearance CL (2.70-6.10 l/h per kg). The pharmacokinetics of NBN were linear and biexponential. Coadministration of BN and NBN resulted in a significant increase in the volume of distribution and clearance of BN. The present results suggest that the nonlinear disposition in the clearance and volume of distribution of BN can be attributed, in part, to the increasing concentration of the metabolite. PMID- 11923062 TI - Fast dispersible ibuprofen tablets. AB - Fast dispersible tablets disintegrate either rapidly in water, to form a stabilized suspension, or disperse instantaneously in the mouth to be swallowed without the aid of water. A direct compression method was used to prepare these two types of tablets containing coated ibuprofen as a high dosed model drug. The properties of the water dispersible tablet, such as porosity, hardness, disintegration time and increase in viscosity after dispersion, were investigated. The selected tablet formulation, containing 26% galactomannan and 5% crospovidone, disintegrates before the galactomannan starts to swell. These tablets disperse in water within 40 s and show a crushing strength of 95 N. To develop an orodispersible tablet, a rotatable central composite design was applied to predict the effects of the quantitative factors mannitol and crospovidone as well as compression force on the characteristics of the tablet. Special emphasis was paid to the development of a wetting test, replacing the normal disintegration method. An optimum tablet formulation, containing 34% mannitol and 13% crospovidone, provides a short wetting time of 17 s and a sufficient crushing strength of 40 N. In conclusion, fast dispersible tablets with acceptable hardness and desirable taste could be prepared within the optimum region. PMID- 11923063 TI - Synthesis of 1,3-thiazine derivatives and their evaluation as potential antimycobacterial agents. AB - A series of eight 5,6-dihydro-4H-1,3-thiazine derivatives was synthesized by the BF3 x Et2O-catalyzed reaction of selected alpha,beta-unsaturated ketones with thiobenzamide at room temperature. The antimycobacterial activities of these compounds were determined against Mycobacterium tuberculosis H37Rv (ATCC 27294) using the Alamar blue susceptibility assay. Three compounds, 5-hydroxy-3-phenyl-4 aza-2-thiabicyclo[3.3.1]none-3-ene 3a, 4-hydroxy-4-methyl-6-pentyl-2-phenyl-5,6 dihydro-4H-1,3-thiazine 3b, and 4-ethyl-4-hydroxy-2-phenyl-5,6-dihydro-4H-1,3 thiazine 3c exhibited inhibitory activities of 97, 77 and 76%, respectively, at a concentration of 6.25 microg/ml. The actual MIC99 for the most active of these compounds, 3a, was also determined to be >6.25 microg/ml. These results, and especially those for 3a, suggest that 1,3-thiazines are potential lead compounds in the search for new antitubercular agents. PMID- 11923064 TI - Cancer and metals and metal compounds: part I--carcinogenesis. PMID- 11923065 TI - The paradox of arsenic: molecular mechanisms of cell transformation and chemotherapeutic effects. AB - Arsenic is a well-documented carcinogen that also appears to be a valuable therapeutic tool in cancer treatment. This creates a paradox for which no unified hypothesis has been reached regarding the molecular mechanisms that determine whether arsenic will act as a carcinogen or as an effectual chemotherapeutic agent. Much of our knowledge with respect to the actions of arsenic has been drawn from epidemiological or clinical studies. The actions of arsenic are likely to be related to cell type, arsenic species, and length and dose of exposure. Arsenic unquestionably induces apoptosis and may specifically target certain tumor cells. Research data strongly suggest that arsenic influences distinct signaling pathways involved in mediating proliferation or apoptosis, including mitogen-activated protein kinases, p53, activator protein-1 or nuclear factor kappa B. The primary purpose of this review is to examine recent findings, from this laboratory and others, that focus on the molecular mechanisms of arsenic's actions in cell transformation and as a therapeutic agent. PMID- 11923066 TI - Carcinogenicity, mutagenicity and teratogenicity of manganese compounds. AB - Manganese, an essential trace element, is one of the most used metals in the industry. Recently, several new manganese compounds have been introduced as fungicide, as antiknock agent in petrol and as contrasting agent in nuclear magnetic resonance tomography. Manganese displays a somewhat unique behaviour with regard to its toxicity. It is relatively non-toxic to the adult organism except to the brain where it causes Parkinson-like symptoms when inhaled even at moderate amounts over longer periods of time. Relatively high doses of manganese affect DNA replication and repair in bacteria and causes mutations in microorganism and mammalian cells although the Ames test does not appear to be particularly responsive to manganese. In mammalian cells, manganese causes DNA damage and chromosome aberrations. Information on organic manganese derivatives is still insufficient. Large amounts of manganese affect fertility in mammals and are toxic to the embryo and foetus. The fungicide MANEB and the contrasting agent MnDPDP also can be embryotoxic, but the latter only at doses much higher than those clinically employed. Information on the anti-knock agent MMT is inadequate. On the other hand, manganese deficiency can also affect fertility and be teratogenic. Information on cancer due to manganese is scanty but the results available do not indicate that inorganic manganese is carcinogenic. More information is desirable with regard to the organic manganese derivatives. It may surprise that an agent that causes mutations is not also carcinogenic. The experience with manganese shows that conclusions with regard to carcinogenicity of an agent based on the observation of mutations are subject to uncertainties. Altogether, it appears that, because of the very high doses at which positive effects have been found, manganese would not represent a significant carcinogenic risk to the population and workers. Care must, however, be exercised with respect to central-nervous symptoms after chronic exposure and with respect to effects on the embryo. Pregnant women should not be exposed to manganese at the work place. PMID- 11923067 TI - Nickel essentiality, toxicity, and carcinogenicity. AB - The increasing utilization of heavy metals in modern industries leads to an increase in the environmental burden. Nickel represents a good example of a metal whose use is widening in modern technologies. As the result of accelerated consumption of nickel-containing products nickel compounds are released to the environment at all stages of production and utilization. Their accumulation in the environment may represent a serious hazard to human health. Among the known health related effects of nickel are skin allergies, lung fibrosis, variable degrees of kidney and cardiovascular system poisoning and stimulation of neoplastic transformation. The mechanism of the latter effect is not known and is the subject of detailed investigation. This review provides an analysis of the current state in the field. PMID- 11923068 TI - Copper and carcinogenesis. AB - Metal ions play an important role in biological systems, and without their catalytic presence in trace or ultratrace amounts many essential co-factors for many biochemical reactions would not take place. However, they become toxic to cells when their concentrations surpass certain optimal (natural) levels. Copper is an essential metal. Catalytic copper, because of its mobilization and redox activity, is believed to play a central role in the formation of reactive oxygen species (ROS), such as O2-* and *OH radicals, that bind very fast to DNA, and produce damage by breaking the DNA strands or modifying the bases and/or deoxyribose leading to carcinogenesis. The chemistry and biochemistry of copper is briefly accounted together with its involvement in cancer and other diseases. PMID- 11923069 TI - The iron metabolism of neoplastic cells: alterations that facilitate proliferation? AB - For many years it has been known that neoplastic cells express high levels of the transferrin receptor 1 (TfR1) and internalize iron (Fe) from transferrin (Tf) at a tremendous rate. Considering the high requirement of neoplastic cells for Fe, understanding its metabolism is vital in terms of devising potential new therapies. Apart from TfR1, a number of molecules have been identified that may have roles in Fe metabolism and cellular proliferation. These molecules include transferrin (Tf), the oestrogen-inducible transferrin receptor-like protein, transferrin receptor 2 (TfR2), melanotransferrin (MTf), ceruloplasmin, and ferritin. In the present review these latter molecules are discussed in terms of their potential functions in tumour cell Fe metabolism and proliferation. Further studies are essential to determine the specific roles of these proteins in the pathogenesis of cancer. PMID- 11923070 TI - Magnesium-DNA interactions and the possible relation of magnesium to carcinogenesis. Irradiation and free radicals. AB - Magnesium deficiency causes renal complications. The appearance of several diseases is related to its depletion in the human body. In radiotherapy, as well as in chemotherapy, especially in treatment of cancers with cis-platinum, hypomagnesaemia is observed. The site effects of chemotherapy that are due to hypomagnesaemia are decreased using Mg supplements. The role of magnesium in DNA stabilization is concentration dependent. At high concentrations there is an accumulation of Mg binding, which induces conformational changes leading to Z DNA, while at low concentration there is deficiency and destabilization of DNA. The biological and clinical consequences of abnormal concentrations are DNA cleavage leading to diseases and cancer. Carcinogenesis and cell growth are also magnesium-ion concentration dependent. Several reports point out that the interaction of magnesium in the presence of other metal ions showed that there is synergism with Li and Mn, but there is magnesium antagonism in DNA binding with the essential metal ions in the order: Zn>Mg>Ca. In the case of toxic metals such as Cd, Ga and Ni there is also antagonism for DNA binding. It was found from radiolysis of deaerated aqueous solutions of the nucleoside 5'-guanosine monophosphate (5'-GMP) in the presence as well as in the absence of magnesium ions that, although the addition of hydroxyl radicals (*OH) has been increased by 2-fold, the opening of the imidazole ring of the guanine base was prevented. This effect was due to the binding of Mg2+ ions to N7 site of the molecule by stabilizing the five-member ring imitating cis-platinum. It was also observed using Fourier Transform Infrared spectroscopy, Raman spectroscopy and Fast Atom Bombardment mass spectrometry that *OH radicals subtract H atoms from the C1', C4' and C5' sites of the nucleotide. Irradiation of 5'-GMP in the presence of oxygen (2.5 x 10(-4) M) shows that magnesium is released from the complex. There is spectroscopic evidence that superoxide anions (O2-*) react with magnesium ions leading to magnesium release from the complex. From radiolysis data it was suggested that magnesium ions can act as radiosensitizers in the absence of oxygen, while in the presence of oxygen they act as protectors and stabilizers of DNA. PMID- 11923071 TI - The role of oxidative stress in mechanisms of metal-induced carcinogenesis. AB - Metals are necessary for the normal functioning of cells and the survival of organisms. However, exposure to higher than the physiological levels of several metals may lead to tumor development. Although the exact molecular mechanism(s) of metal-induced carcinogenesis is not clear, a vast body of evidence indicates that metal-induced generation of reactive oxygen species (ROS) may play a central role in this process. Two main pathways of ROS-induced effects are discussed in this chapter: (i) increased DNA damage induced either directly or indirectly by impeding DNA repair, and (ii) modulation of nuclear transcriptional factor activities, such as NF-kappaB and AP-1, through mitogen-activated protein kinases signal transduction mechanisms. PMID- 11923072 TI - Intracellular signal transduction of cells in response to carcinogenic metals. AB - Epidemiological and animal studies suggest that several metals and metal containing compounds are potent mutagens and carcinogens. These metals include chromium, arsenic, vanadium, nickel, and others. During the last two decades, chemical and cellular studies have contributed enormously to our understanding of the mechanisms of metal-induced pathophysiological processes. Although each of these metals is unique in its mechanism of action, some common signaling molecules, such as reactive oxygen species (ROS), may be shared by many of the carcinogenic metals. New techniques are now available to reveal the mechanisms of carcinogenesis in precise molecular terms. In this review, we focused our attentions on carcinogenic metal-induced signal transduction pathways leading to the activation of NF-kappaB, cell apoptosis and cell cycle progression, three crucial steps or events involved in the transformation and carcinogenesis. This review summarizes current knowledge and our recent studies concerning intracellular signal transduction pathways initiated by carcinogenic metals and the cross-talk that occurs among these pathways in cells in response to metals. PMID- 11923073 TI - The ecophysiology of air-breathing in crabs with special reference to Gecarcoidea natalis. AB - To succeed on land rather than in water, crabs require a suite of physiological and morphological changes, and ultimately the ability to reproduce without access open water. Some species have modified gills to assist in gas exchange but accessory gas exchange organs, usually lungs, occur in many species. In accomplished air-breathers the lung becomes larger and more vascularised with pulmonary vessels directing oxygenated haemolymph to the heart. The relative abundance of O(2) in air promotes relative hypoventilation and thus an internal hypercapnia to drive CO(2) excretion. Land crabs have a dual circulation via either lungs or gills and shunting between the two may depend on respiratory media or exercise state. During their breeding migration on Christmas Island Gecarcoidea natalis maintained arterial Po(2) by branchial O(2) uptake, while pulmonary O(2) pressure was reduced; partly because exercise doubled relative haemolymph flow through the gills. Related species rely on elevated haemocyanin concentration and affinity for O(2) to assist uptake but this compromises unloading at the tissues and thus the aerobic scope of tissues. Aquatic crabs exchange salt and ammonia with water via the gills but in land crabs this is not possible. Birgus latro has adopted uricotelism but other species excrete ammonia in either the urine or as gas. Land crabs minimise urinary salt loss using a filtration-reabsorption system analogous to the kidney. Urine is redirected across the gills where salt reabsorption occurs in systems under hormonal control, although in G. natalis this is stimulatory and in B. latro inhibitory. While crabs occupy a range of habitats from aquatic to terrestrial, these species do not comprise a physiological continuum but across the crab taxa individual species possess appropriate and specific physiological features to survive in their individual habitat. PMID- 11923075 TI - Physiological variation in insects: large-scale patterns and their implications. AB - In this paper we demonstrate how broad scale comparative physiology has an important role to play in informing a variety of assumptions made in macroecology. We do so by examining large-scale geographic variation in insect development, thermal tolerance and metabolic rate. From these studies, and those from the literature on insect water loss and thermoregulation, we show that there is often a bias to the geographic extent of available empirical data. Studies of cold hardiness are most usually undertaken at high latitudes, while investigations of upper thermal tolerances and water loss are most common in warm arid regions. Likewise, we demonstrate that much variation in insect physiological tolerances is partitioned at higher taxonomic levels, which has important implications for comparative physiology. Intriguingly, data on the full range of variables we review are available for only three species. We also show that, despite its importance, body size is regularly reported in only some kinds of investigations (metabolic rate, water loss rate), whereas in others (upper lethal temperature, cold hardiness, development) this variable is often ignored. In short, although large-scale comparative physiology can contribute considerable understanding to both physiology and ecology, there is much that remains to be done. PMID- 11923074 TI - Adaptive heterothermy and selective brain cooling in arid-zone mammals. AB - Adaptive heterothermy and selective brain cooling are regarded as important thermal adaptations of large arid-zone mammals. Adaptive heterothermy, a process which reduces evaporation by storing body heat, ought to be enhanced by ambient heat load and by water deficit, but most mammals studied fail to show at least one of those attributes. Selective brain cooling, the reduction of brain temperature below arterial blood temperature, is most evident in artiodactyls, which possess a carotid rete, and traditionally has been considered to protect the brain during hyperthermia. The development of miniature ambulatory data loggers for recording body temperature allows the temperatures of free-living wild mammals to be measured in their natural habitats. All the African ungulates studied so far, in their natural habitats, do not exhibit adaptive heterothermy. They have low-amplitude nychthemeral rhythms of temperature, with mean body temperature over the night exceeding that over the day. Those with carotid retes (black wildebeest, springbok, eland) employ selective brain cooling but zebra, without a rete, do not. None of the rete ungulates, however, seems to employ selective brain cooling to prevent the brain overheating during exertional hyperthermia. Rather, they use it at rest, under moderate heat load, we believe in order to switch body heat loss from evaporative to non-evaporative routes. PMID- 11923076 TI - The timing of hibernation in Tasmanian echidnas: why do they do it when they do? AB - We investigated the patterns of hibernation and arousals in seven free-ranging echidnas Tachyglossus aculeatus setosus (two male, five female) in Tasmania using implanted temperature data loggers. All echidnas showed a 'classical' pattern of mammalian hibernation, with bouts of deep torpor interrupted by periodic arousals to euthermia (mean duration 1.04+/-0.05 (n=146). Torpor bout length increased as body temperature fell during the hibernation season, and became more variable as temperature rose again. Hibernation started in late summer (February 28+/-5 days, n=6) and males aroused just before the winter solstice (June 15+/-3 days, n=3), females that subsequently produced young aroused 40 days later (July 25+/-3, n=4) while females that did not produce young hibernated for a further two months (arousal Sept 27+/-5, n=7). We suggest that hibernation in Tasmanian echidnas can be divided into two phases, the first phase, marked by declining minimum body temperatures as ambient temperature falls, appears to be obligatory for all animals, while the second phase is 'optional' and is utilised to varying amounts by females. We suggest that early arousal and breeding is the favoured option for females in good condition, and that the ability to completely omit breeding in some years, and hibernate through to spring is an adaptation to an uncertain climate. PMID- 11923077 TI - Xylose as a nectar sugar: from biochemistry to ecology. AB - Studies of nectar sugar composition in the Proteaceae, an ancient southern hemisphere plant family, have demonstrated that xylose comprises up to 39% of nectar sugar in two genera, Protea and Faurea, and may therefore represent a substantial fraction of the energy available to pollinators of these plants. Although insect and bird pollinators of Protea species are averse to xylose, mice (Aethomys namaquensis) will drink pure xylose, which is metabolized either by gut bacteria or by the mouse tissues. In the form of xylan polymers, the pentose sugar D-xylose is a structural component of plant cell walls, and there is considerable biotechnological interest in xylose fermentation. Bacteria and yeasts convert D-xylose to D-xylulose and thence via the pentose phosphate pathway to fructose-6-phosphate, which is either oxidized or fermented to ethanol. Gut symbionts of rodent pollinators may be analogous to ruminal xylose metabolizing bacteria. The presence of xylose in Protea and Faurea nectar remains puzzling in view of pollinator aversions: even for rodent pollinators, it is the least preferred nectar sugar. In the generalized pollination systems of the Proteaceae, a coevolutionary explanation for nectar xylose as an attractant for mammalian pollinators is probably less likely than one involving plant physiology, with xylose in phloem sap being secreted passively into the nectar. PMID- 11923078 TI - Birth in marsupials. AB - Birth is an event that allows the relatively immature marsupial to move from the internal environment of the uterus to the external environment of the pouch. The newborn marsupial passes down from the uterus to the urogenital sinus and then makes its way to the pouch and attaches to the teat at a very early stage of development. From the studies available, there appear to be three methods used by the newborn to move from the uterus to the pouch. In marsupials with a forward pouch such as the red kangaroo, tammar wallaby and the brushtail possum, the mother positions her urogenital sinus below the pouch and the newborn climb upward towards the pouch. The young climb with a swimming motion, moving the head from side-to-side and use the forearms in alternate strokes. In the bandicoot with a backward facing pouch, the mother positions the urogenital sinus above the pouch and the young slither down into the pouch. The young do not have a definite crawl, as seen with the macropodids and possum. The third method of birth has been observed in the marsupials without a definite pouch that have a mammary region that develops as the young grow in size. This type of pouch is observed in the dasyurids. The mother was noted to stand on four legs with her hips raised so that the urogenital sinus was above the pouch and the newborn young crawled downwards from the sinus to the pouch. In all species, birth was completed in 2-4 min. PMID- 11923079 TI - Evolution of viviparity: what can Australian lizards tell us? AB - Historically, Australia has been important in the study of, and the development of hypotheses aimed at understanding, the evolution of viviparity in amniote vertebrates. Part of the importance of Australia in the field results from a rich fauna of skinks, including one of the broadest ranges of diversity of placental structures within one geographic region. During the last decade, we have focussed our studies on one lineage, the Eugongylus group of skinks of the subfamily Lygosominae because it contains oviparous species and some that exhibit complex placentae. Our specific objective has been to attempt to understand the fundamental steps required when viviparity, and ultimately complex placentae, evolve from oviparous ancestors. We have taken a three-prong approach: (1) detailed study of the morphology and ontogeny of the placentae of key species at the light microscope level; (2) study of changes in the uterus associated with pregnancy, or the plasma membrane transformation; and (3) measures of the net exchange of nutrients across the placenta or eggshell of key species. In turn, we have found that: (1) details of the morphology and ontogeny of placentae are more complex that originally envisaged, and that the early conclusions about a sequence in the evolution of complex placentae was naive; (2) a plasma membrane transformation occurs in viviparous, but not oviparous lizards, and thus may be a fundamental feature of the evolution of viviparity in amniotes; and (3) species with more complex chorioallantoic placentae tend to transport more nutrients across the placenta during pregnancy than those with simpler chorioallantoic placentae but, because the correlation is not tight, the importance of the omphaloplacenta in transporting nutrients may have been overlooked. Also, the composition of yolk of highly matrotrophic species is broadly similar, but not identical, to the yolk of oviparous species. Some of the interpretation of our data within the context of our specific objective is not yet possible, pending the publication of a robust phylogeny of Eugongylus group skinks. Once such a phylogeny is available, we are in a position to propose specific hypotheses about the evolution of viviparity that can be tested using another lineage of amniotes, possibly Mabuya group skinks. PMID- 11923080 TI - Pollination by passerine birds: why are the nectars so dilute? AB - Bird-pollinated flowers are known to secrete relatively dilute nectars (with concentrations averaging 20-25% w/w). Many southern African plants that are pollinated by passerine birds produce nectars with little or no sucrose. Moreover, these hexose nectars are extremely dilute (10-15%). This suggests a link between sugar composition and nectar concentration. Nectar originates from sucrose-rich phloem sap, and the proportion of monosaccharides depends on the presence and activity of invertase in the nectary. Hydrolysis of sucrose increases nectar osmolality and the resulting water influx can potentially convert a 30% sucrose nectar into a 20% hexose nectar, with a 1.56 times increase in volume. Hydrolysis may also increase the gradient for sucrose transport and thus the rate of sugar secretion. When sucrose content and refractometer data were compared, some significant correlations were seen, but the occurrence of sucrose-rich or hexose-rich nectars can also be explained on phylogenetic grounds (e.g. Erythrina and Protea). Hexose nectars may be abundant enough to drip from open flowers, but evaporation leads to much variability in nectar concentration and increases the choices available to pollinators. PMID- 11923081 TI - Brain-specific lipids from marine, lacustrine, or terrestrial food resources: potential impact on early African Homo sapiens. AB - The polyunsaturated fatty acid (PUFA) composition of the mammalian central nervous system is almost wholly composed of two long-chain polyunsaturated fatty acids (LC-PUFA), docosahexaenoic acid (DHA) and arachidonic acid (AA). PUFA are dietarily essential, thus normal infant/neonatal brain, intellectual growth and development cannot be accomplished if they are deficient during pregnancy and lactation. Uniquely in the human species, the fetal brain consumes 70% of the energy delivered to it by mother. DHA and AA are needed to construct placental and fetal tissues for cell membrane growth, structure and function. Contemporary evidence shows that the maternal circulation is depleted of AA and DHA during fetal growth. Sustaining normal adult human brain function also requires LC PUFA.Homo sapiens is unlikely to have evolved a large, complex, metabolically expensive brain in an environment which did not provide abundant dietary LC-PUFA. Conversion of 18-carbon PUFA from vegetation to AA and DHA is considered quantitatively insufficient due to a combination of high rates of PUFA oxidation for energy, inefficient and rate limited enzymatic conversion and substrate recycling. The littoral marine and lacustrine food chains provide consistently greater amounts of pre-formed LC-PUFA than the terrestrial food chain. Dietary levels of DHA are 2.5-100 fold higher for equivalent weights of marine fish or shellfish vs. lean or fat terrestrial meats. Mammalian brain tissue and bird egg yolks, especially from marine birds, are the richest terrestrial sources of LC PUFA. However, land animal adipose fats have been linked to vascular disease and mental ill-health, whereas marine lipids have been demonstrated to be protective. At South African Capesites, large shell middens and fish remains are associated with evidence for some of the earliest modern humans. Cape sites dating from 100 to 18 kya cluster within 200 km of the present coast. Evidence of early H. sapiens is also found around the Rift Valley lakes and up the Nile Corridor into the Middle East; in some cases there is an association with the use of littoral resources. Exploitation of river, estuarine, stranded and spawning fish, shellfish and sea bird nestlings and eggs by Homo could have provided essential dietary LC-PUFA for men, women, and children without requiring organized hunting/fishing, or sophisticated social behavior. It is however, predictable from the present evidence that exploitation of this food resource would have provided the advantage in multi-generational brain development which would have made possible the advent of H. sapiens. Restriction to land based foods as postulated by the savannah and other hypotheses would have led to degeneration of the brain and vascular system as happened without exception in all other land based apes and mammals as they evolved larger bodies. PMID- 11923082 TI - The effects of the deep-sea environment on transmembrane signaling. AB - Membrane-associated processes may be particularly susceptible to perturbation by the high hydrostatic pressures and low temperatures of the deep ocean. Transmembrane signaling by guanyl nucleotide binding protein (G protein) coupled receptors (GPCRs) is affected at a number of steps: (1) agonist activation of the GPCR; (2) the interaction of the receptor with the heterotrimeric G protein; (3) the G protein GTPase cycle; and (4) the activation and function of the effector element, adenylyl cyclase. The effects of low temperature and high hydrostatic pressures on the A(1) adenosine receptor-inhibitory G protein (G(i))-adenylyl cyclase signaling complex were examined in teleost fishes from three families, Scorpaenidae, Macrouridae and Moridae. In a comparison of teleost fishes, rat and chicken, species with body temperatures from 1 to 40 degrees C, at atmospheric pressure, A(1) adenosine receptor agonist binding is conserved at the body temperature of the species. In the marine teleost fishes examined, increased pressure decreases agonist efficacy. There are differences among species in the effects of increased hydrostatic pressure on G protein interactions with receptors, GTP binding to G protein alpha subunits and the intrinsic GTPase activity of alpha subunits. Adenylyl cyclase activity and modulation are affected by increased pressure in all the species examined, except Antimora rostrata which was unaffected by pressure changes. At pressures approximating those which the species experience in situ adenylyl cyclase activity retains its sensitivity to modulators. To understand the physiological consequences of impaired cell signaling several prototypical human diseases are discussed. PMID- 11923083 TI - Comparison of growth and lipid composition in the green abalone, Haliotis fulgens, provided specific macroalgal diets. AB - Lipid composition of abalone was examined over a one-year interval. A feeding trial was designed to cover a full reproductive cycle in young adult green abalone, Haliotis fulgens, consisting of five diet treatments: the macrophytic algal phaeophyte Egregia menziesii, rhodophyte Chondracanthus canaliculatus, chlorophyte Ulva lobata, a composite of the three algae and a starvation control. The lipid class, fatty acid, sterol and 1-O-alkyl glyceryl ether profiles were determined for foot, hepatopancreas/gonad tissues and larvae. The major fatty acids were 16:0, 18:0, 18:1(n-7)c, 18:1(n-9)c, 20:4(n-6), 20:5(n-3) and 22:5(n 3), as well as 14:0 for abalone fed brown and red algae. 4,8,12 Trimethyltridecanoic acid, derived from algae, was detected for the first time in H. fulgens (hepatopancreas complex, 1.2-13.9%; larvae, 0.5% of total fatty acids). Diacylglyceryl ethers were present in larvae (0.6% of total lipid). The major 1-O-alkyl glycerols were 16:0, 16:1 and 18:0. Additionally, 18:1(n-9) was a major component in hepatopancreas/gonad and larvae. The major sterol was cholesterol (96-100% of total sterols). Highest growth rates were linked to temperature and occurred in abalone fed the phaeophyte E. menziesii (43 microm.day(-1), 56 mg.day(-1) yearly mean), an alga containing the highest levels of C(20) polyunsaturated fatty acids and the highest ratio of 20:4(n-6) to 20:5(n 3). This study provides evidence of the influence of diet and temperature on seasonal changes in abalone lipid profiles, where diet is most strongly related to body mass and temperature to shell length. The allocation of lipids to specific tissues in green abalone clarifies their lipid metabolism. These results provide a basis for improving nutrition of abalone in mariculture through formulation of artificial feeds. PMID- 11923084 TI - Myofibril-bound serine protease and its endogenous inhibitor in mouse: extraction, partial characterization and effect on myofibrils. AB - The protein content of muscle is determined by the relative rates of synthesis and degradation. The balance between this process determines the number of functional contractile units within each muscle cell. Myofibril-bound protease, protease M previously reported in mouse skeletal muscle could be solubilized from the myofibrillar fraction by salt and acid treatment and partially purified by Mono Q and Superose 12 chromatography. Isolated protease M activity in vitro on whole myofibrils resulted in myosin, actin, troponin T, alpha-actinin and tropomyosin degradation. Protease M is serine type and was able to hydrolyze trypsin-type synthetic substrates but not those of chymotrypsin type. In gel filtration chromatography, protease M showed Mr 120.0 kDa. The endogenous inhibitor (MHPI) is a glycoprotein (110.0 kDa) that efficiently blocks the protease M-dependent proteolysis of myofibrillar proteins in a dose-dependent way, as shown by electrophoretic analysis and synthetic substrates assays. Protease M-Inhibitor system would be implicated in myofibrillar proteins turnover. PMID- 11923085 TI - Identification of two distinct E-NTPDases in liver of goldfish (Carassius auratus L.). AB - We have recently reported the existence of ATPase activity capable of hydrolyzing extracellular ATP and localized at the external cell membrane of goldfish hepatocytes [Am. J. Physiol. (1998) 274 R1031]. In the present study, we investigated whether one or more enzymes of the ATP diphosphohydrolase family (called E-NTPDases) are responsible for the hydrolysis of extracellular ATP and other nucleotides. Using soluble extracts from goldfish liver, enzyme activity was detected in the presence of ATP (32.1+/-4.0 nmol Pi liberated mg protein(-1) min(-1)), ADP (20.7+/-3.3 nmol Pi liberated mg protein(-1) min(-1)) and UTP (20.7+/-1.2 nmol Pi liberated mg protein(-1) min(-1)). In line with the presence of this hydrolytic activity, liver samples separated by non-denaturing gel electrophoresis and subsequently exposed to either ATP, ADP or UTP yielded a single band with enzyme activity and similar electrophoretic mobility. Subsequent SDS-PAGE electrophoresis of the active bands resulted in the appearance of two protein bands with molecular masses of 70 and 64 kDa. Immunoblotting of soluble extracts and microsomes obtained from goldfish liver, using a monoclonal antibody against CD39 (a well-known E-NTPDase), detected a single 97-kDa protein. The enzyme activity measured in solution and in native gels, together with structural information from denaturing gels plus immunoblots, points to the existence, in goldfish liver, of at least two different E-NTPDases. PMID- 11923086 TI - Interannual and between species comparison of the lipids, fatty acids and sterols of Antarctic krill from the US AMLR Elephant Island survey area. AB - Antarctic euphausiids, Euphausia superba, E. tricantha, E. frigida and Thysanoessa macrura were collected near Elephant/ Island / during 1997 and 1998. Total lipid was highest in E. superba small juveniles (16 mg g(-1) wet mass), ranging from 12 to 15 mg in other euphausiids. Polar lipid (56-81% of total lipid) and triacylglycerol (12-38%) were the major lipids with wax esters (6%) only present in E. tricantha. Cholesterol was the major sterol (80-100% of total sterols) with desmosterol second in abundance (1-18%). 1997 T. macrura and E. superba contained a more diverse sterol profile, including 24 nordehydrocholesterol (0.1-1.7%), trans-dehydrocholesterol (1.1-1.5%), brassicasterol (0.5-1.7%), 24-methylenecholesterol (0.1-0.4%) and two stanols (0.1-0.2%). Monounsaturated fatty acids included primarily 18:1(n-9)c (7-21%), 18:1(n-7)c (3-13%) and 16:1(n-7)c (2-7%). The main saturated fatty acids in krill were 16:0 (18-29%), 14:0 (2-15%) and 18:0 (1-13%). Highest eicosapentaenoic acid [EPA, 20:5(n-3)] and docosahexaenoic acid [DHA, 22:6(n-3)] occurred in E. superba (EPA, 15-21%; DHA, 9-14%), and were less abundant in other krill. E. superba is a good source of EPA and DHA for consideration of direct or indirect use as a food item for human consumption. Lower levels of 18:4(n-3) in E. tricantha, E. frigida and T. macrura (0.4-0.7% of total fatty acids) are more consistent with a carnivorous or omnivorous diet as compared with herbivorous E. superba (3.7 9.4%). The polyunsaturated fatty acid (PUFA) 18:5(n-3) and the very-long chain (VLC-PUFA), C(26) and C(28) PUFA, were not present in 1997 samples, but were detected at low levels in most 1998 euphausiids. Interannual differences in these biomarkers suggest greater importance of dinoflagellates or some other phytoplankton group in the Elephant Island area during 1998. The data have enabled between year comparisons of trophodynamic interactions of krill collected in the Elephant Island region, and will be of use to groups using signature lipid methodology. PMID- 11923087 TI - Precursors of three unique cysteine-rich peptides from the scorpion Buthus martensii Karsch. AB - Scorpion venoms contain a large number of small peptides with diverse primary structures and unique pharmacological functions. From a cDNA library prepared from venom glands of the Chinese scorpion Buthus martensii Karsch, clones encoding precursors of three unique cysteine-rich peptides named BmTXKS3, BmTXLP2 and BmAP1 have been isolated and sequenced. These precursors are composed of 54, 94 and 89 amino acids, respectively, containing a signal peptide in their N termini. Sequence analysis shows that BmTXKS3 and BmTXLP2 are two novel members of a scorpion toxin family sharing cysteine-stabilized alpha-helical folds. BmAP1 possesses a distinctive cystine framework, which is similar to some serine protease inhibitors and the segments of several extracellular proteins. PMID- 11923088 TI - Characteristics of protectant synthesis of infective juveniles of Steinernema carpocapsae and importance of glycerol as a protectant for survival of the nematodes during osmotic dehydration. AB - Two hypotheses on the synthesis of the protectants glycerol and trehalose of the infective juveniles (IJs) of Steinernema carpocapsae during osmotic dehydration were tested and utilised to evaluate the function and importance of glycerol on survival of the nematodes during osmotic dehydration. This was achieved by comparing the changes in survival, morphology, behaviour and levels of glycerol, trehalose and permeated compounds of the IJs dehydrated in seven hypertonic solutions at two temperature regimes: (1) 5 degrees C for 15 days; and (2) 23 degrees C for 1 day followed by 5 degrees C for another 14 days. The results substantiate both hypotheses tested: (1) the permeability of the IJs to various compounds, such as sucrose or ethylene glycol, when they are dehydrated in hypertonic solutions of these compounds; and (2) suppression of the synthesis of protectant glycerol but not trehalose when IJs are dehydrated at low temperature. The results also showed that: (1) although trehalose was the preferred dehydration protectant, glycerol played an important role in rapidly balancing the osmotic pressure when IJs were exposed in hypertonic solutions; (2) the presence of glycerol was essential for the IJs to survive and function properly even under moderate osmotic dehydration, especially when IJs were dehydrated in salt solutions; and (3) some exogenous compounds permeated into IJs during osmotic dehydration such as ethylene glycol, may function in the same way as glycerol and significantly improve the survival and function of the IJs. The results indicate that each of the protectants glycerol and trehalose has a specific function and neither is replaceable by the other. PMID- 11923089 TI - Separation of highly fluorescent proteins by SDS-PAGE in Acroporidae corals. AB - This study characterized the spectral properties of Acropora tenuis, A. nasuta, A. secale, and A. aspera, all of which showed strong colorful fluorescence under ultraviolet light-A (black light). The emission maxima of fluorescence from the intact corals were 517, 482, 484, and 514 nm in A. tenuis, A. nasuta, A. secale, and A. aspera, respectively. Using a soluble fraction of cell-free extract of the corals, we applied a method of sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) to separate each fluorescent protein component contained in the corals. Green fluorescent bands were detected in all Acropora examined, although their apparent molecular mass and relative content were different. A. aspera had two orange bands in addition to the green one. The major excitation and emission peaks of the orange fluorescence bands were almost identical (476 and 478 nm), however, they were discernible by their spectral profiles and molecular masses. Some biochemical properties of the highly fluorescent proteins of Acropora are described and implications of the results are discussed. PMID- 11923090 TI - Organization of the lipoprotein lipase gene of red sea bream Pagrus major. AB - Lipoprotein lipase (LPL) is a key enzyme of lipid deposition and metabolism. To investigate the mechanism of lipid deposition in fish, as a first step, we have characterized the LPL gene of a marine teleost red sea bream Pagrus major by cDNA and genomic structure analysis. The red sea bream LPL gene encodes 511 amino acids and spans approximately 6.3 kb of the genome. The coding region is organized into ten exons and nine introns. In comparison with the LPL of other animals, the deduced amino acid sequence shows a high degree of similarity with a conservation of functional domains, e.g. catalytic triad, N-glycosylation sites, lipid and heparin binding regions. The 1.1 kb of 5' flanking region contains two CCAAT, sequences homologous to Oct-I site and response elements for hormones including glucocorticoid, insulin and thyroid hormone. The results of the present study will facilitate further study of the function and regulation of the LPL in non-mammalian vertebrates. PMID- 11923091 TI - Sulfatides and arylsulfatase A activity in major salivary glands of hamster (Mesocricetus auratus) after adenocarcinoma induction in oral cavity. AB - A biochemical study of sulfatides and arylsulfatase A (ASA) was carried out in the submandibular and sublingual glands of the male and female hamster Mesocricetus auratus after experimental induction of oral adenocarcinoma by 7,12 dimethylbenzanthracene (DMBA). Hamster experimental groups included control animals, animals treated with beta-carotene, animals treated with DMBA, and animals treated with DMBA plus beta-carotene. Oral cavity treatment with DMBA induced carcinogenesis in the buccal mucosa, but not in the major salivary glands, where nevertheless, the morphology and expression of both parameters examined changed. In fact, sulfatide concentrations and enzyme activity increased significantly, while in control and beta-carotene-treated hamsters they were similar in both glands and sexes. After administration of DMBA plus beta carotene, sulfatide concentration decreased, as did ASA activity, slightly in the submandibular gland and remarkably so in the sublingual one of female hamsters. Thin-layer chromatography (TLC) analysis of lipid patterns, after DMBA treatment, revealed considerable differences, not only in sulfatides, but also in other lipid fractions, as well as between the two glands and two sexes. These findings show that oral cavity treatment with DMBA is not able to induce carcinogenesis in the major salivary glands examined; however, it does cause considerable metabolic changes. PMID- 11923092 TI - Expression, purification and DNA-binding activity of tilapia muscle-specific transcription factor, MyoD, produced in Escherichia coli. AB - MyoD is one of several helix-loop-helix proteins regulating muscle-specific gene expression. Using a reverse transcription-polymerase chain reaction, 5'-rapid cDNA end amplification, and plaque hybridization, MyoD cDNA was cloned from the mRNA of tilapia dorsal skeletal muscle. The 1015 bp MyoD cDNA product contained an 846 bp open reading frame with flanking regions of 115 and 64 bp at the 5'- and 3'-ends, respectively. Results showed that the tilapia MyoD sequence, which includes one polypeptide of 281 amino acids, shared sequence identities of 64.3, 64.1, 62.6 and 62.4% with those of zebrafish, carp, and two rainbow trout, respectively. Results from a molecular phylogenic tree assay showed that the tilapia MyoD was more closely related to those of other fishes than of higher vertebrates. Using Escherichia coli, a pET expression system, and an Ni(2+)-NTA column, we purified approximately 35 kDa recombinant tilapia MyoD. Results from an electrophoretic mobility shift assay demonstrated that the purified E. coli produced tilapia MyoD was capable of binding to the DNA fragment sequence CA(C/T)(C/A)TG. PMID- 11923093 TI - Sciatic nerve transection decrease substance P immunoreactivity in the lumbosacral spinal cord of the frog (Rana catesbeiana). AB - Using immunohistochemistry and optical densitometry, substance P (SP) was investigated in the lumbar spinal cord of the frog Rana catesbeiana after sciatic nerve transection. In control animals, there was a high density of SP fibers in the Lissauer's tract and in the mediolateral band of the dorsal gray matter. Other SP immunoreactive fibers were observed in the dorsal part of the lateral funiculus and in the ventral horn. No SP label was found in any cell bodies. After axotomy, SP immunoreactive fibers decreased in the Lissauer's tract on the same side of the lesion. The other regions remained labeled. The changes were observed at 3 days following axonal injury and persisted at 5, 8 and 15 days. At 20 days, there was no significant difference between the axotomized side and the control one, thus indicating a recovery of the SP expression. These results indicate that the frog may be used as a model to study the effects of peripheral axotomy, contributing to elucidate the SP actions in the pain neuropath. PMID- 11923094 TI - Role of antioxidant systems in induced nutritional pancreatic atrophy in chicken. AB - One-day-old chicks were reared using diets differing in their vitamin E and/or selenium content. The purpose of this research was to detect any possible imbalance in the antioxidant defense system, which could be related to development of nutritional pancreatic atrophy. Mitochondrial membranes from animals deficient in both nutrients, or just vitamin E, submitted to peroxidizability 'in vitro' had the production of TBARS greatly enhanced. Measurements of the 2-GSH/GSSG ratio suggested that selenium and vitamin E, the latter in higher magnitude, were responsible for maintenance of the reducing capacity of the cell. Enzymatic defense systems against oxidative stress were also studied. The results indicated that the total antioxidant enzymatic activity of pancreatic cells was not sufficient to scavenge all the ROS generated in the nutritionally deficient animals. The present study suggests that nutritional deficiency of selenium and/or vitamin E generates one imbalance between pro oxidant and antioxidant systems in chicken pancreas, leading to oxidative stress and pancreatic atrophy. PMID- 11923095 TI - 1-O-Hexadecyl-2-desoxy-2-amino-sn-glycerol, a substrate for human sphingosine kinase. AB - The substrate specificity of human sphingosine kinase was investigated using a bacterially expressed poly(His)-tagged protein. Only the D-erythro isomer of the sphingoid bases, sphinganine and sphingenine, was effectively phosphorylated. Long chain 1-alkanols, alkane-1,2-diols, 2-amino-1-alkanol or 1-amino-2-alkanol and short chain 2-amino-1,3-alkanediols were very poor substrates, indicating that the kinase is recognizing the chain length and the position of the amino and secondary hydroxy group. A free hydroxy group at carbon 3 is not a prerequisite, however, since 1-O-hexadecyl-2-desoxy-2-amino-sn-glycerol was an efficient substrate with an apparent K(m) value of 3.8 microM (versus 15.7 microM for sphingenine). This finding opens new perspectives to design sphingosine kinase inhibitors. It also calls for some caution since it cannot be excluded that this ether lipid analogue is formed from precursors that are frequently used in research on platelet activating factor or from phospholipid analogues which are less prone to degradation. PMID- 11923097 TI - Active taurocholic acid flux through hepatoma cells increases the cellular pool of unesterified cholesterol derived from lipoproteins. AB - The effect of bile acid flux on the fate of lipoprotein-derived cholesterol was studied in bile acid-transporting McNtcp.18 hepatoma cells. The intracellular unesterified cholesterol (UC) concentration rose when McNtcp.18 cells grown in the presence of either high density lipoproteins (HDL) or low density lipoproteins (LDL) were incubated with taurocholic acid (TCA). This effect was more pronounced when the exogenous source of cholesterol was HDL. The presence of TCA in the culture medium of McNtcp.18 cells had no discernible effect on the uptake of cholesteryl esters (CE) from either lipoprotein. TCA treatment of cells preincubated with either lipoprotein did not affect cholesterol synthesis but antagonized the stimulation of cholesterol esterification in cells that were incubated with LDL. The CE concentration in cells treated with TCA was decreased, relative to cells not incubated with TCA, suggesting that cellular CE stores were also hydrolyzed. The TCA treatment reduced the amount of total cholesterol released into the medium by the lipoprotein-treated cells, which was coincident with the reduction in the amount of apolipoprotein B in the culture medium. However, the proportion of UC released into the medium by the lipoprotein-treated cells was increased in cells capable of active bile acid transport. The results indicate that active bile acid flux through hepatoma cells increases the cellular pool of UC derived from lipoproteins. The UC released by the cells into the culture medium under this condition may represent cholesterol destined for direct biliary secretion. PMID- 11923096 TI - Functional implications of post-translational modifications of phospholipases D1 and D2. AB - Our previous studies showed that truncation of the N-terminal 168 amino acids of rat brain phospholipase D1 (rPLD1) abolishes its response to protein kinase C (PKC) and greatly diminishes its palmitoylation and Ser/Thr phosphorylation. In this study, we show that the response to PKC as well as the palmitoylation and Ser/Thr phosphorylation were restored when the truncated rPLD1 mutant (rPLD1(DeltaN168)) was coexpressed with a fragment containing the N-terminal 168 amino acids. Immunoprecipitation experiments showed that the N-terminal fragment associated with rPLD1(DeltaN168) when coexpressed in COS 7 cells and that palmitoylation of Cys(240) and Cys(241) was not necessary for the association. In addition, we found that rat PLD2 (rPLD2) was palmitoylated on Cys(223) and Cys(224) in COS 7 cells. Mutation of both these cysteines reduced the basal activity of rPLD2, however its response to PMA stimulation in vivo was retained. As in the case of rPLD1, loss of palmitoylation weakened membrane association of rPLD2. In summary, the N-terminal 168-amino-acid fragment of rPLD1 can associate with truncated rPLD1(DeltaN168) to restore its palmitoylation, Ser/Thr phosphorylation and PKC response. Although rPLD2 differs from rPLD1 in many properties, it is palmitoylated at the corresponding conserved cysteine residues in COS 7 cells. PMID- 11923098 TI - Comparative cytotoxic and cytoprotective effects of taurohyodeoxycholic acid (THDCA) and tauroursodeoxycholic acid (TUDCA) in HepG2 cell line. AB - This study was performed to compare the effects of two hydrophilic bile acids, taurohyodeoxycholic acid (THDCA) and tauroursodeoxycholic acid (TUDCA), on HepG2 cells. Cytotoxicity was evaluated at different times of exposure by incubating cells with increasing concentrations (50-800 micromol/l) of either bile acid, while their cytoprotective effect was tested in comparison with deoxycholic acid (DCA) (350 micromol/l and 750 micromol/l)-induced cytotoxicity. Culture media, harvested at the end of each incubation period, were analyzed to evaluate aspartate transaminase (AST), alanine transaminase and gamma glutamyltranspeptidase release. In addition, the hemolytic effect of THDCA and TUDCA on human red blood cells was also determined. At 24 h of incubation neither THDCA nor TUDCA was cytotoxic at concentrations up to 200 and 400 micromol/l. At 800 micromol/l both THDCA and TUDCA induced a slight increase in AST release. At this concentration and with time of exposure prolonged up to 72 h, THDCA and TUDCA induced a progressive increase of AST release significantly (P<0.05) higher than that of controls being AST values for THDCA (2.97+/-0.88 time control value (tcv) at 48 h and 4.50+/-1.13 tcv at 72 h) significantly greater than those of TUDCA (1.50+/-0.20 tcv at 48 h and 1.80+/-0.43 tcv at 72 h) (P<0.01). In cytoprotection experiments, the addition of 50 micromol/l THDCA decreased only slightly (-5%) AST release induced by 350 micromol/l DCA, while the addition of 50 micromol/l TUDCA was significantly effective (-23%; P<0.05). Higher doses of THDCA or TUDCA did not reduce toxicity induced by 350 micromol/l DCA, but were much less toxic than an equimolar dose of DCA alone. At the concentration used in this experimental model neither THDCA nor TUDCA was hemolytic; however at a very high concentration (6 mmol/l) both bile acids induced 5-8% hemolysis. We conclude that bile acid molecules with a similar degree of hydrophilicity may show different cytotoxic and cytoprotective properties. PMID- 11923099 TI - Delta6-, Stearoyl CoA-, and Delta5-desaturase enzymes are expressed in beta-cells and are altered by increases in exogenous PUFA concentrations. AB - In the evolution of Type II diabetes, an initial period of hyper-fatty acidemia leads to an insulin secretory defect which triggers overt hyperglycemia and frank diabetes. The mechanism by which elevated free fatty acids contribute to beta cell dysfunction, however, is not clearly understood. We recently reported that arachidonic acid (20:4) or linoleic acid (18:2) supplementations result in increases in abundances of long chain polyunsaturated fatty acids in INS-1 beta cell membrane lipids, suggesting that beta-cells express desaturases that catalyze generation of unsaturated fatty acids. As expression of desaturases by beta-cells has not yet been addressed, we initiated studies to examine this issue using INS-1 beta-cells and find that they express messages for the Delta6-, stearoyl CoA-, and Delta5-desaturase. Supplementation of the INS-1 beta-cells with arachidonic acid leads to decreased expression of all three desaturases, presumably in response to the decreased need for endogenous generation of unsaturated fatty acids. In contrast, linoleic acid supplementation promoted minimal changes in the three desaturases. These findings demonstrate for the first time that beta-cells express regulatable desaturases. Additionally, reverse transcriptase-polymerase chain reaction analyses reveal expression of the desaturases in native pancreatic islets. It might be speculated that long-term elevations in fatty acids can also adversely influence desaturase activity in beta-cells and affect PUFA composition in beta-cell membranes contributing to beta-cell membrane structural abnormalities and altered secretory function. PMID- 11923100 TI - Torpor-associated fluctuations in surfactant activity in Gould's wattled bat. AB - The primary function of pulmonary surfactant is to reduce the surface tension (ST) created at the air-liquid interface in the lung. Surfactant is a complex mixture of lipids and proteins and its function is influenced by physiological parameters such as metabolic rate, body temperature and breathing. In the microchiropteran bat Chalinolobus gouldii these parameters fluctuate throughout a 24 h period. Here we examine the surface activity of surfactant from warm-active and torpid bats at both 24 degrees C and 37 degrees C to establish whether alterations in surfactant composition correlate with changes in surface activity. Bats were housed in a specially constructed bat room at Adelaide University, at 24 degrees C and on a 8:16 h light:dark cycle. Surfactant was collected from bats sampled during torpor (2535 degrees C). Alterations in the lipid composition of surfactant occur with changes in the activity cycle. Most notable is an increase in surfactant cholesterol (Chol) with decreases in body temperature [Codd et al., Physiol. Biochem. Zool. 73 (2000) 605 612]. Surfactant from active bats was more surface active at higher temperatures, indicated by lower ST(min) and less film area compression required to reach ST(min) at 37 degrees C than at 24 degrees C. Conversely, surfactant from torpid bats was more active at lower temperatures, indicated by lower ST(min) and less area compression required to reach ST(min) at 24 degrees C than at 37 degrees C. Alterations in the Chol content of bat surfactant appear to be crucial to allow it to achieve low STs during torpor. PMID- 11923101 TI - Both isoforms of mammalian phosphatidylinositol transfer protein are capable of binding and transporting sphingomyelin. AB - The structurally related mammalian alpha and beta isoforms of phosphatidylinositol (PtdIns) transfer protein (PITP) bind reversibly a single phospholipid molecule, preferably PtdIns or phosphatidylcholine (PtdCho), and transport that lipid between membrane surfaces. PITPbeta, but not PITPalpha, is reported extensively in the scientific literature to exhibit the additional capacity to bind and transport sphingomyelin (CerPCho). We undertook a detailed investigation of the lipid binding and transfer specificity of the soluble mammalian PITP isoforms. We employed a variety of donor and acceptor membrane lipid compositions to determine the sensitivity of recombinant rat PITPalpha and PITPbeta isoforms toward PtdIns, PtdCho, CerPCho, and phosphatidate (PtdOH). Results indicated often striking differences in protein-phospholipid and protein membrane interactions. We demonstrated unequivocally that both isoforms were capable of binding and transferring CerPCho; we confirmed that the beta isoform was the more active. The order of transfer specific activity was similar for both isoforms: PtdIns>PtdCho>CerPCho>>PtdOH. Independently, we verified the binding of CerPCho to both isoforms by showing an increase in holoprotein isoelectric point following the exchange of protein-bound phosphatidylglycerol for membrane associated CerPCho. We conclude that PITPalpha and PITPbeta are able to bind and transport glycero- and sphingophospholipids. PMID- 11923102 TI - The identification of intestinal scavenger receptor class B, type I (SR-BI) by expression cloning and its role in cholesterol absorption. AB - The molecular mechanisms of cholesterol absorption in the intestine are poorly understood. With the goal of defining candidate genes involved in these processes a fluorescence-activated cell sorter-based, retroviral-mediated expression cloning strategy has been devised. SCH354909, a fluorescent derivative of ezetimibe, a compound which blocks intestinal cholesterol absorption but whose mechanism of action is unknown, was synthesized and shown to block intestinal cholesterol absorption in rats. Pools of cDNAs prepared from rat intestinal cells enriched in enterocytes were introduced into BW5147 cells and screened for SCH354909 binding. Several independent clones were isolated and all found to encode the scavenger receptor class B, type I (SR-BI), a protein suggested by others to play a role in cholesterol absorption. SCH354909 bound to Chinese hamster ovary (CHO) cells expressing SR-BI in specific and saturable fashion and with high affinity (K(d) approximately 18 nM). Overexpression of SR-BI in CHO cells resulted in increased cholesterol uptake that was blocked by micromolar concentrations of ezetimibe. Analysis of rat intestinal sections by in situ hybridization demonstrated that SR-BI expression was restricted to enterocytes. Cholesterol absorption was determined in SR-B1 knockout mice using both an acute, 2-h, assay and a more chronic fecal dual isotope ratio method. The level of intestinal cholesterol uptake and absorption was similar to that seen in wild type mice. When assayed in the SR-B1 knockout mice, the dose of ezetimibe required to inhibit hepatic cholesterol accumulation induced by a cholesterol containing 'western' diet was similar to wild-type mice. Thus, the binding of ezetimibe to cells expressing SR-B1 and the functional blockade of SR-B1-mediated cholesterol absorption in vitro suggest that SR-B1 plays a role in intestinal cholesterol metabolism and the inhibitory activity of ezetimibe. In contrast studies with SR-B1 knockout mice suggest that SR-B1 is not essential for intestinal cholesterol absorption or the activity of ezetimibe. PMID- 11923103 TI - IL-2 mediates adjuvant effect of dendritic cells. AB - Research into the biology of dendritic cells (DCs) has shown that they play a central role in priming early and late immune responses. The recent finding that DCs produce interleukin-2 (IL-2) after microbial challenge suggests that this could be the mechanism by which DCs link innate and adaptive immunity. This hypothesis is supported also by the observation that murine cytomegalovirus, which is able to establish a persistent infection, can interfere with the ability of infected DCs to produce IL-2 and prime T cells. PMID- 11923104 TI - Cross-priming versus cross-tolerance: are two signals enough? AB - Depending on their state of maturation, dendritic cells (DCs) can cross-prime or cross-tolerize T cells. Mature DCs provide T-cell-receptor ligands (signal 1) and costimulatory molecules (signal 2), and activate T cells. Immature DCs provide signal 1, but not signal 2, and tolerize T cells. However, new data show that it is not the expression of costimulatory molecules alone that determines whether DCs induce immunity or tolerance, because mature DCs can induce tolerance also. PMID- 11923105 TI - Challenges and issues in new vaccine development. AB - The Euroconference on Vaccines of the Future: from Rational Design to Clinical Development was held at the Pasteur Institute, Paris, France from 17-19 October 2001. PMID- 11923106 TI - Telomere-based therapy for immunosenescence. AB - The International Workshop on Telomeres and the Immune System: Ageing and Novel Intervention Strategies was held at Birmingham University, UK from 1-3 December 2001. PMID- 11923111 TI - Immunoneutralization and anti-idiotype production: two-sided applications of leptin. AB - The neuroendocrine and immune systems are linked through a complex bi-directional network, in which hormones modify immune function, and the immune system, through the action of cytokines, affects neuroendocrine responses involved in the maintenance of body homeostasis. The adipocyte-derived, peptide hormone leptin is a pleiotropic molecule belonging to the helical cytokine family. On pp. 182-187, Matarese et al. suggest the possibility of new leptin-based therapeutic strategies for the treatment of both infection and autoimmune disease. PMID- 11923112 TI - Balancing susceptibility to infection and autoimmunity: a role for leptin? AB - The immune responses to many infections have long been known to share features with autoimmune responses. In particular, both types of response are typified by the enhanced reactivity of T helper 1 cells - with high levels of interleukin-2, interferon gamma and tumor necrosis factor alpha - and are accompanied often by organ-specific and/or systemic damage and the production of IgG. Paradoxically, the geographical distributions of incidence of infectious diseases and autoimmunity are complementary, rather than coincident. In less-developed societies, an epidemiological association between susceptibility to infection and malnutrition has been observed, whereas in affluent countries, an increased incidence of autoimmune diseases has been described. We suggest that these observations can be explained partly by taking into consideration the immune effects of the adipocyte-derived hormone leptin, which has been shown recently to act as a link between nutritional status and the immune response. PMID- 11923113 TI - Defining the dynamics of self-assembled Fas-receptor activation. AB - Fas-Fas-ligand, the most well investigated apoptotic signaling system, plays a pivotal role in several human diseases. Its signaling through multiple pathways; multiple, complex regulatory points; and recently described tendency to undergo ligand-independent self-assembly warrant a comprehensive understanding of its physiologically well controlled function. In this review, we attempt to develop such a perspective on the regulation of Fas signaling. The multistep Fas regulation model presented here should be helpful in devising effective interventions for conditions resulting from the dysregulation of Fas. PMID- 11923114 TI - The importance of lytic and nonlytic immune responses in viral infections. AB - Antiviral immune effector mechanisms can be divided broadly into lytic and nonlytic components. We use mathematical models to investigate the fundamental question of which type of response is required to combat different types of viral infection. According to our model, the relative roles of the two types of component depend on the cytopathicity of the virus relative to its rate of replication. If the viral cytopathicity is low relative to the rate of viral replication, the model predicts that a combination of lytic and nonlytic effector mechanisms is likely to be required to resolve the disease, particularly if the virus replicates at a fast rate. By contrast, if viral cytopathicity is high relative to the replication rate of the virus, then lytic and nonlytic mechanisms can, in principle, resolve the infection independently. We discuss our findings in the context of specific viral infections and use our model to interpret empirical data. PMID- 11923115 TI - Cytokines as a link between innate and adaptive antitumor immunity. AB - Recent studies indicate that cytokines produced by cells of the innate defense system play an essential role in influencing the immune response towards protective antitumor immunity. These cytokines might act as first 'danger signals' in alerting the immune system. By promoting the differentiation and activation of dendritic cells, antigen presentation and T-cell-mediated immune responses, these cytokines could be powerful natural adjuvants for the development of cancer vaccines. With regard to type I interferons - cytokines with a long history of clinical use - this new knowledge could be translated easily into novel strategies for the prevention and treatment of human malignancies. PMID- 11923116 TI - Stimulation of local antibody production: parenteral or mucosal vaccination? AB - Mucosal vaccines aim to prevent the penetration of pathogens, including HIV, into the body. Their cost can be low compared with curative therapies and thus, they are well suited to the fight against pandemic diseases in developing countries. Efficacy has been demonstrated for the oral poliovirus vaccine, but only very few other vaccines administered by the mucosal route are available commercially at present. Tremendous research efforts have now improved significantly the classical approach of these vaccines, and alternative methods of immunization, based on new concepts of mucosal immunity, are being developed. PMID- 11923117 TI - Antifertility vaccines. AB - The possibility of using immunization as a method of birth control has been explored actively since the 1930s, with several different sperm, egg or hormonal antigens having been studied as suitable targets for intervention. However, it is only in the past decade that the efficacy of vaccination against fertility has become established firmly in both humans and free-roaming animal populations. We will review recent progress in the continuing development of antifertility vaccines, with an emphasis on vaccines intended ultimately for use in humans, whilst highlighting also some of the notable successes achieved with vaccines produced for use in other species. PMID- 11923118 TI - Future prospects for vaccines to control fertility. AB - Vaccination is used routinely to protect against infectious disease and is being explored increasingly as a method of protection against tumors. Also, it has been established that vaccination using antigens associated with reproduction can protect against undesired pregnancy. Substantial progress over the past decade suggests that, if the remaining immunological and socioeconomic issues can be resolved, antifertility vaccines could be a valuable, additional method of family planning. PMID- 11923121 TI - Shifting the LDL-receptor paradigm in familial hypercholesterolemia: novel insights from recent kinetic studies of apolipoprotein B-100 metabolism. AB - Familial hypercholesterolemia (FH) is a dominantly inherited disorder associated with elevated plasma cholesterol concentrations and premature cardiovascular disease. In addition to impaired low density lipoprotein (LDL) receptor-mediated clearance of low density lipoproteins in FH, evidence from in vitro and in vivo studies suggests that hepatic oversecretion of apoB may contribute to the hypercholesterolemia. The proposed association between apoB secretion and FH may, however, be a function of the class of LDL receptor defect. Hepatic cholesterol pools appear to regulate apoB secretion and LDL receptor activity. Therefore, therapeutic regulation of cholesterogenesis in FH may have the dual effect of reducing hepatic apoB secretion and upregulating the LDL receptor. These effects may also be genetically determined. PMID- 11923122 TI - Environmental modulation of atherosclerosis end points in familial hypercholesterolemia. AB - In familial hypercholesterolemia (FH), early coronary heart disease (CHD) is a complex trait that results from a large monogenic component of susceptibility due to elevated LDL cholesterol. This was demonstrated by observation of the high risk of early CHD in FH subjects compared with the general population. However, not all subjects with a LDLR gene mutation suffer with early CHD. Furthermore, studies in extended multigenerational families showed that even for this strong monogenic effect the environment could substantially modulate the age at death from CHD. Anecdotal examples of apparent modulation of atherosclerosis severity by lifestyle changes were also seen in other monogenic metabolic problems, such as hepatic lipase deficiency and Dunnigan-type familial partial lipodystrophy. Thus, even within apparently clear-cut rare monogenic metabolic diseases, such as FH, among carriers there can be a variability in the expression of important quantitative end points, such as early CHD. In some cases, the environment, including lifestyle factors, appears to play a key role in modulating the disease severity. This complexity could have implications for diagnosis and treatment. PMID- 11923123 TI - MEDPED and the Spanish Familial Hypercholesterolemia Foundation. AB - We analysed clinical and genetic data of 819 non-related familial hypercholesterolemia patients. No differences on baseline low density lipoprotein (LDL) cholesterol levels between females and males were observed. High density lipoprotein (HDL) cholesterol values were higher in females than in males (57+/ 15 vs. 47.7+/-13, respectively, P<0.01). Premature cardiovascular disease was present in 21.7% of cases (30.8% in males and 14.3% in females, P<0.001). A significant and positive correlation was observed between cardiovascular disease and age, gender, tobacco, total and LDL cholesterol levels at diagnosis, and negatively with HDL cholesterol levels. Analysis of LDL-receptor gene have been performed in 350 cases and 86 different mutations have been found. PMID- 11923124 TI - The management of dysplastic Barrett's esophagus: where do we go from here? PMID- 11923125 TI - Utilization of sentinel lymph node mapping to determine pathologic outcomes for patients receiving neoadjuvant chemotherapy for locally advanced breast cancer. PMID- 11923126 TI - Adjuvant chemoradiotherapy in gastric cancer: wave goodbye to extensive surgery? PMID- 11923127 TI - High-grade dysplasia within Barrett's esophagus: controversies regarding clinical opinions and approaches. AB - Barrett's esophagus with high-grade dysplasia is a well-known risk factor for the development of esophageal adenocarcinoma, which has become the predominant form of esophageal cancer in the United States. This review addresses four major fundamental issues that shape our treatment decisions regarding high-grade dysplasia within Barrett's esophagus: (1) the poorly defined natural history of high-grade dysplasia in its progression to adenocarcinoma, (2) the potentially high morbidity and mortality of esophageal resection for high-grade dysplasia, (3) the difficulty in detecting cancer among dysplastic cells during endoscopy, and (4) the controversial role of endoscopic mucosal ablative therapy for high grade dysplasia. Until there are more accurate surveillance methods, better biochemical or molecular markers in predicting cancerous progression, or more effective minimally invasive methods of treatment, esophagogastrectomy must be considered the standard means of managing patients with Barrett's esophagus and high-grade dysplasia. Regular rigorous systematic surveillance and endoscopic mucosal ablation are alternative treatment options that are available but should be used only under strict conditions. The decision to proceed in a certain direction is quite complex and challenging and ideally requires the feedback of patients who are properly educated about the controversies surrounding this disease. PMID- 11923128 TI - A prospective trial of preoperative chemotherapy in resectable breast cancer: predictors of breast-conservation therapy feasibility. AB - BACKGROUND: The role of preoperative chemotherapy for breast cancer is evolving. We initiated a prospective trial of sequential preoperative paclitaxel and doxorubicin-based combination chemotherapy in patients with stage I (tumor >1 cm), II, or IIIA disease and evaluated its effect on breast-conservation therapy (BCT) eligibility. METHODS: Pathology findings for the initial 100 consecutive patients who underwent surgery were analyzed. RESULTS: The median tumor size at presentation was 2.4 cm, and 39% of patients were deemed eligible for BCT. After chemotherapy, the median tumor size decreased to 1.0 cm (P <.001), and 59% of patients seemed BCT eligible (BCT conversion rate 34% among patients initially assessed as BCT ineligible; P <.001). Final pathology confirmed BCT feasibility in 90% of patients assessed as BCT candidates before surgery. The pathology from mastectomy specimens revealed BCT feasibility in 11 (27%) of 41 patients deemed BCT ineligible. Multivariate analysis revealed lobular histology, multicentricity, and calcifications, but not age, initial tumor size, or nodal status to predict final pathology indicating BCT ineligibility. CONCLUSIONS: Induction chemotherapy improves BCT eligibility for breast cancer patients. Improved breast imaging methods after chemotherapy are necessary to improve accuracy in predicting the feasibility of BCT, especially in patients presenting with diffuse calcifications or multicentricity. PMID- 11923129 TI - Sentinel lymphadenectomy after neoadjuvant chemotherapy for breast cancer may reliably represent the axilla except for inflammatory breast cancer. AB - BACKGROUND: After neoadjuvant chemotherapy, women with locally advanced breast cancer (LABC) undergo a modified radical mastectomy or lumpectomy with axillary lymph node dissection (ALND) and radiotherapy. Sentinel lymphadenectomy (SL) is accepted for axillary evaluation in early breast cancer. We assessed the feasibility and predictive value of SL after neoadjuvant chemotherapy. METHODS: Eligible women received neoadjuvant therapy for LABC and were scheduled to undergo a definitive surgical procedure. Vital blue dye SL was attempted followed by level I and II axillary dissection. RESULTS: SL was successful in 29 of 34 patients (detection rate, 85%). Thirteen patients (45%) had positive nodes, and eight (28%) had negative nodes on both SL and ALND. In five patients (17%), the sentinel node was the only positive node identified. Overall, there was a 90% concordance between SL and ALND. The false-negative rate and negative predictive value were 14% and 73%, respectively. Among the subgroup without inflammatory cancer, the detection and concordance rates were 89% and 96%, respectively. The false-negative rate was 6%, and the negative predictive value was 88%. CONCLUSIONS: SL after neoadjuvant chemotherapy may reliably predict axillary staging except in inflammatory breast cancer. Further studies are required to assess the utility of SL as the only mode of axillary evaluation in these women. PMID- 11923130 TI - Analysis of sentinel lymph node mapping with immediate pathologic review in patients receiving preoperative chemotherapy for breast carcinoma. AB - BACKGROUND: Sentinel lymph node mapping (SLNM) and neoadjuvant chemotherapy are becoming established components of therapy for selected patients with breast carcinoma. However, neoadjuvant therapy has been considered a relative contraindication to SLNM. In an effort to learn whether patients who have received preoperative chemotherapy can undergo accurate SLNM, we evaluated our experience with this technique. METHODS: From January 1997 to June 2000, SLNM and axillary lymph node dissection were concurrently performed in 35 patients who received preoperative chemotherapy. Mapping was performed with (99m)Tc sulfur colloid only in one patient and Lymphazurin dye only in 15 patients, and the two methods were combined in the remainder. RESULTS: SLNM successfully identified a sentinel lymph node in 30 (86%) patients. Metastatic disease was identified in the sentinel lymph nodes of four patients during surgery. The intraoperative pathologic diagnosis proved to be correct in 19 (79%) of 24 patients. The final pathologic diagnosis of the sentinel lymph node reflected the status of the axillary contents in all patients in whom it was identified. CONCLUSIONS: These results demonstrate that SLNM can be consistently performed in patients receiving preoperative chemotherapy for breast cancer, suggesting the utility of this technique in this patient population. PMID- 11923131 TI - Final results of the Department of Defense multicenter breast lymphatic mapping trial. AB - BACKGROUND: Lymphatic mapping and sentinel lymph node (SLN) biopsy have the potential to become the standard of care for nodal staging in breast cancer patients, but their widespread utility outside of university-based centers has not been determined. This study describes the final results from a national multi institutional trial designed to determine the role of preoperative lymphoscintigraphy in breast lymphatic mapping, the rate of success for finding an SLN, and the rate of skip metastasis for patients with invasive breast cancer across all practice scenarios. METHODS: Lymphatic mapping techniques involving the combined use of blue dye and radiocolloid were taught to participating surgeons through a formal 2-day training course at the Moffitt Cancer Center. In protocol 1, surgeons performed their first 20 to 25 cases of breast mapping with SLN biopsy followed by complete axillary lymph node dissection. In protocol 2, after the learning phase, surgeons did not perform axillary lymph node dissection unless a SLN was positive for metastatic disease. RESULTS: Forty-two institutions, including 12 university-based research centers, participated in the trial. From July 1, 1997, through January 31, 1999, a total of 965 patients were accrued. Lymphoscintigraphy identified drainage to an axillary SLN 64% of the time, but by using sensitive handheld gamma probes at the time of the operation, an axillary SLN could be identified 86% of the time. The rate of success for finding an axillary SLN was 92.8% for cases performed at the Moffitt Cancer Center. For other university centers, the rate of success of identifying an axillary SLN was 91.4%, and for other community/regional hospitals in the study, it was 85.2%. For cases in which protocol 1 was followed, the rate of false negative SLN biopsy was 4%. There was no axillary nodal recurrence after a negative SLN in protocol 2 when a negative SLN biopsy was followed by observation. The median follow-up for the patients on protocol 2 was 16 months. CONCLUSIONS: These data show a high rate of success for finding an axillary SLN and a low rate of skip metastasis in a national multicenter study of lymphatic mapping for breast cancer. This study suggests that SLN biopsy for breast cancer can be performed successfully in community/regional hospitals, as well as in major university-based centers. PMID- 11923132 TI - Predictors of locoregional recurrence among patients with early-stage breast cancer treated with breast-conserving therapy. AB - BACKGROUND: Our aim was to identify predictors of locoregional recurrence (LRR) in patients with early-stage breast cancer treated with breast-conserving therapy (BCT) and long-term follow-up. METHODS: From 1970 to 1994, 1153 patients with stage I to II breast cancer underwent BCT and radiotherapy at our institution. Patients with prior breast cancer or other primary malignancies were excluded. Clinical and pathologic characteristics evaluated were age, race, tumor size, stage, pathologic tumor margins, axillary nodal involvement, estrogen and progesterone receptor status, Black's nuclear grade, type of surgery, and use of adjuvant therapy. RESULTS: Of 1083 patients, 54% presented with stage I disease and 46% with stage II disease. Median age was 50 years, and median follow-up was 9 years. Axillary nodes were positive in 31% of the patients who underwent axillary dissection. LRR developed in 6%, LRR followed by systemic recurrence in 5%, and systemic recurrence alone in 13%; 76% had no evidence of recurrence at last follow-up. Age, tumor size, positive lymph nodes, and not receiving chemotherapy or hormonal therapy were independent predictors of LRR. Disease specific survival among patients with LRR was similar to that among patients with no recurrence. CONCLUSIONS: Multidisciplinary treatment strategies should be used to accomplish durable locoregional control after BCT. PMID- 11923133 TI - Pathologic and biological prognostic factors of breast cancers in short- and long term hormone replacement therapy users. AB - BACKGROUND: Breast cancer tumors occurring in hormone replacement therapy (HRT) users are less aggressive, but no studies have compared tumor aggressiveness among HRT users by length and mode of therapy. METHODS: A total of 1105 consecutive postmenopausal patients treated for operable breast cancer at the European Institute of Oncology were identified. Women exposed to HRT were compared with HRT nonusers by clinical stage at presentation and pathologic and biological tumor characteristics. HRT duration and administration modality were analyzed in relation to tumor characteristics in the HRT group. RESULTS: Better stage distribution, including smaller pathologic tumor diameter and fewer involved axillary lymph nodes, was seen in the HRT group. Estrogen receptor positive tumors were more frequent in the control group, but this tendency was reversed with longer exposure to HRT. Histological grade III tumors were less frequent in the HRT group. More favorable prognostic factors were associated with HRT >5 years. The proliferative fraction was higher in patients with exposure <1 vs. >5 years and in oral versus transdermal users. CONCLUSIONS: Breast cancers developing during HRT have better prognostic characteristics than those seen in HRT nonusers. A trend toward better prognostic characteristics with increasing duration of HRT was seen. PMID- 11923134 TI - The effect of prior breast biopsy method and concurrent definitive breast procedure on success and accuracy of sentinel lymph node biopsy. AB - BACKGROUND: It has been suggested that sentinel lymph node (SLN) biopsy for breast cancer may be less accurate after excisional biopsy of the primary tumor compared with core needle biopsy. Furthermore, some have suggested an improved ability to identify the SLN when total mastectomy is performed compared with lumpectomy. This analysis was performed to determine the impact of the type of breast biopsy (needle vs. excisional) or definitive surgical procedure (lumpectomy vs. mastectomy) on the accuracy of SLN biopsy. METHODS: The University of Louisville Breast Cancer Sentinel Lymph Node Study is a prospective multi-institutional study. Patients with clinical stage T1-2, N0 breast cancer were eligible. All patients underwent SLN biopsy and completion level I/II axillary dissection. Statistical comparison was performed by chi(2) analysis. RESULTS: A total of 2206 patients were enrolled in the study. There were no statistically significant differences in SLN identification rate or false negative rate between patients undergoing excisional versus needle biopsy. The SLN identification and false-negative rates also were not statistically different between patients who had total mastectomy compared with those who had a lumpectomy. CONCLUSIONS: Excisional biopsy does not significantly affect the accuracy of SLN biopsy, nor does the type of definitive surgical procedure. PMID- 11923136 TI - Poor prognosis associated with thrombocytosis in patients with gastric cancer. AB - BACKGROUND: Thrombocytosis is commonly associated with malignant disease and has recently been suggested to be a poor prognostic indicator in patients with lung cancer and gynecological cancers. The prevalence of thrombocytosis in patients with gastric cancer was reviewed, and its association with poor prognosis was investigated. METHODS: Platelet count (PLT) and hemoglobin concentrations (Hb) were reviewed in 369 consecutive patients with histologically verified gastric cancer from 1994 to 2000. Differences between categories were analyzed with analysis of variance, and survival was compared by using the log-rank test on the Kaplan-Meier life table. Multivariate Cox regression analysis was used to evaluate whether thrombocytosis is an independent prognostic marker. RESULTS: Thrombocytosis was found in 42 patients, and anemia was found in 200 patients. PLT was negatively correlated with Hb. Mean PLT was significantly increased in patients with noncurative operations. There was a positive correlation between the depth of tumor invasion and PLT. One- and 3-year survival expectancies in patients with or without thrombocytosis were 52.4% and 23.4% and 85.7% and 72.9%, respectively. PLT was identified as an independent prognostic factor after lymph node metastasis and depth of tumor invasion. CONCLUSIONS: Thrombocytosis is an independent prognostic indicator of survival in patients with gastric cancer. PMID- 11923135 TI - Surgical treatment variation in a prospective, randomized trial of chemoradiotherapy in gastric cancer: the effect of undertreatment. AB - BACKGROUND: Intergroup 0116 (Southwest Oncology Group 9008), a national, multicenter, two-armed, prospective, randomized trial of adjuvant postoperative chemoradiotherapy, has demonstrated significant benefit. METHODS: We prospectively captured complete surgical information, including the treatment of various lymph node stations, for 553 of the 556 eligible participants in this trial. Before any survival analysis, we coded D level by using the Japanese general rules and used the Maruyama program to estimate the likelihood of disease in undissected regional node stations, defining the sum of these estimates as the Maruyama Index of Unresected Disease (MI). We analyzed survival with Cox multivariate regression. RESULTS: Fifty-four percent of participating patients underwent D0 lymphadenectomy. The median MI was 70 (range, 0-429). In contrast to D level, MI proved to be an independent prognostic factor, even with adjustment for the potentially linked variables of T stage and number of positive nodes. We detected no significant interaction between surgical or pathologic variables and the favorable effect of adjuvant treatment, but the power to detect such interaction was generally low. CONCLUSIONS: MI, a measure of unresected regional nodal disease in gastric cancer, proved an independent predictor of survival. Surgical undertreatment, as observed in this trial, clearly undermined survival. PMID- 11923138 TI - Primary thyroid lymphoma: can the diagnosis be made solely by fine-needle aspiration? AB - BACKGROUND: Primary malignant lymphoma of the thyroid accounts for <5% of all thyroid malignancies and is primarily treated with chemotherapy and external beam radiation. With the advent of modern immunophenotypic analyses, fine-needle aspiration (FNA) can potentially obviate the need for surgical procedures. METHODS: To investigate the utility of FNA, data from 23 consecutive patients with primary malignant thyroid lymphoma evaluated at the Johns Hopkins Hospital from July 1985 to April 2000 were analyzed. RESULTS: Patients were categorized into two groups: those diagnosed before 1993 (group 1, n = 12) and those diagnosed after 1993 (group 2, n = 11). Although patients in group 1 were slightly older, there were no other differences between the groups with regard to sex, tumor grade, or tumor stage. Although no patient in group 1 was successfully diagnosed by FNA alone, seven patients (63%) in group 2 were diagnosed solely by FNA (P =.019, chi(2) analysis). Therefore, all 12 patients in group 1, but only 4 of 11 patients in group 2, required open surgical biopsy. CONCLUSIONS: Primary thyroid lymphoma is an uncommon malignancy usually treated nonsurgically once the diagnosis is established. In most patients with malignant lymphoma of the thyroid, FNA, should obviate the need for open surgical biopsy. PMID- 11923137 TI - An experimental evaluation of three preoperative radiation regimens for resectable rectal cancer. AB - BACKGROUND: We investigated the degree of tumor cell killing after radiotherapy regimens commonly used in clinical practice in comparison with an accelerated schedule. METHODS: Mtln3 mammary adenocarcinoma tumor cells were inoculated subcutaneously in the hind leg of syngeneic Fischer 344 rats. Tumors were irradiated with 5 x 5 Gy in 5 days, 10 x 3 Gy over 10 days, or 5 x (2 x 3) Gy in 5 days. After excision of the irradiated tumors, the dye exclusion, a tetrazolium based colorimetric and the clonogenic assays were used to determine tumor cell viability and surviving fractions. RESULTS: Estimated potential doubling time values indicate a rapid proliferation capacity, comparable with potential doubling time values in human rectal cancer. The dye exclusion and clonogenic assays revealed a significantly higher degree of cell killing after the hypofractionated and the accelerated regimens of, respectively, 5 x 5 Gy and 5 x (2 x 3) Gy over 5 days compared with 10 x 3 Gy over 10 days. CONCLUSIONS: A shorter treatment time offered the best therapeutic efficacy. The schedule involving two daily fractions of 3 Gy over 5 days should be less toxic than 5 x 5 Gy and may therefore provide a therapeutic advantage. PMID- 11923139 TI - Adverse effects of the antiangiogenic agent angiostatin on the healing of experimental colonic anastomoses. AB - BACKGROUND: Antiangiogenic cancer therapy is likely to be administered long term for sustained suppression of tumor outgrowth. Surgeons will encounter more patients undergoing such therapy. Therefore, it is essential to know the effects of antiangiogenic agents on physiological angiogenesis, as occurs during the healing of colonic anastomoses. METHODS: Angiostatin was generated from human plasma and administered continuously. In 38 mice, the right colon was anastomosed after transection: group 1 (n = 13), anastomotic healing under angiostatin treatment from surgery until death (day 7); group 2 (n = 13), phosphate-buffered saline controls. For healing on discontinuation of treatment, group 3 (n = 6) received angiostatin treatment preceding surgery during 4 days; group 4 (n = 6) included controls. On day 7, all mice were inspected for signs of anastomotic leakage. Bursting pressure measurements were performed to test anastomotic strength. Neovascularization was assessed semiquantitatively by immunohistochemistry. RESULTS: Mice treated with angiostatin postoperatively showed significantly more signs of leakage, more adhesions, and peritonitis. One mouse died on day 5. Five mice had paralytical ileus. The bursting pressure in group 1 was 135 +/- 20 mm Hg, versus 175 +/- 12 mm Hg in group 2 (mean +/- SEM). Significantly fewer new vessels were found surrounding the anastomosis in the treated group (6.6 +/-.9) versus controls (16 +/- 1.6). All controls, as well as those animals treated with angiostatin only until surgery (group 3), displayed normal healing and showed no signs of peritonitis or ileus. CONCLUSIONS: Angiostatin impairs anastomotic healing in mice. However, on discontinuation of antiangiogenic therapy, normal anastomotic healing is promptly restored. PMID- 11923140 TI - Phosphatase and tensin analog gene overexpression engenders cellular death in human malignant mesothelioma cells via inhibition of AKT phosphorylation. AB - BACKGROUND: Abnormal phosphatase and tensin analog (PTEN) gene expression has been noted in neoplasms. The PTEN protein cleaves phosphate groups from cellular growth kinases, inhibiting tumor propagation. A downstream target of PTEN is AKT, a serine-threonine kinase that when activated inhibits apoptosis. We sought to determine whether PTEN overexpression in mesothelioma cells would engender hypophosphorylation of AKT and apoptosis. METHODS: Human malignant mesothelioma cell lines REN and I-45 were transfected with adenoviral vectors AdPTEN and AdBgal (marker gene) at various multiplicities of infection (MOI). Cell viability was measured using a colorimetric assay, and apoptosis was assessed by morphology and subG1 fluorescence-activated cell sorter (FACS) analysis. PTEN protein and AKT phophorylation were evaluated by Western blot, and AKT kinase activity was evaluated by functional assay. RESULTS: Increased cellular killing was noted with AdPTEN gene transfer. The ratio of cell killing with AdPTEN to AdBgal widened with increasing MOI and was statistically significant at all MOI. Cells demonstrated apoptosis by morphologic and subG1 FACS analyses. Cells overexpressing PTEN demonstrated decreased phosphorylated (active) AKT and AKT kinase activity compared with controls. CONCLUSIONS: Overexpression of PTEN engenders apoptosis in mesothelioma by AKT hypophosphorylation. The forced overexpression of PTEN may prove useful clinically in this treatment-resistant neoplasm. PMID- 11923141 TI - Comment on the article "Highest isotope count does not predict sentinel node positivity in all breast cancer patients," by Martin et al., August 2001, Annals of Surgical Oncology. PMID- 11923142 TI - Initial antiretroviral regimens. PMID- 11923143 TI - The long case versus objective structured clinical examinations. PMID- 11923144 TI - Falling neonatal autopsy rates. PMID- 11923145 TI - Protein conjugate pneumococcal vaccines. PMID- 11923146 TI - Global nursing shortages. PMID- 11923147 TI - "Unlawfully treated" woman to move to new unit. PMID- 11923148 TI - Pfizer gets a public dressing down over promoting unlicensed drugs. PMID- 11923150 TI - Transplants from live patients scrutinised after donor's death. PMID- 11923151 TI - BMA negotiator calls for more male medical students. PMID- 11923152 TI - Cancer money siphoned off to pay debts. PMID- 11923153 TI - New test simplifies genetic testing for breast cancer. PMID- 11923154 TI - Overall health of Hungary's Roma worse than average population. PMID- 11923156 TI - Paediatricians meet to tackle child abuse in former Soviet bloc. PMID- 11923157 TI - Systematic review and meta-analysis of evidence for increasing numbers of drugs in antiretroviral combination therapy. AB - OBJECTIVE: To assess the evidence for the effectiveness of increasing numbers of drugs in antiretroviral combination therapy. DESIGN: Systematic review, meta analysis, and meta-regression of fully reported randomised controlled trials. All studies included compared quadruple versus triple therapy, triple versus double therapy, double versus monotherapy, or monotherapy versus placebo or no treatment. PARTICIPANTS: Patients with any stage of HIV infection who had not received antiretroviral therapy. MAIN OUTCOME MEASURES: Changes in disease progression or death (clinical outcomes); CD4 count and plasma viral load (surrogate markers). SEARCH STRATEGY: Six electronic databases, including Medline, Embase, and the Cochrane Library, searched up to February 2001. RESULTS: 54 randomised controlled trials, most of good quality, with 66 comparison groups were included in the analysis. For both the clinical outcomes and surrogate markers, combinations with up to and including three (triple therapy) were progressively and significantly more effective. The odds ratio for disease progression or death for triple therapy compared with double therapy was 0.6 (95% confidence interval 0.5 to 0.8). Heterogeneity in effect sizes was present in many outcomes but was largely related to the drugs used and trial quality. CONCLUSIONS: Evidence from randomised controlled trials supports the use of triple therapy. Research is needed on the effectiveness of quadruple therapies and the relative effectiveness of specific combinations of drugs. PMID- 11923158 TI - Ten years of neonatal autopsies in tertiary referral centre: retrospective study. AB - OBJECTIVES: To measure the neonatal autopsy rate at a tertiary referral centre and identify trends over the past decade. To identify factors that may influence the likelihood of consent being given for autopsy. To examine any discordance between diagnoses before death and at autopsy. DESIGN: Retrospective review of patients' records. SETTING: Tertiary neonatal referral centre affiliated to university. OUTCOME MEASURES: Sex, gestational age, birth weight, type of delivery, and length of stay in neonatal unit for baby. Maternal age, marital status, history of previous pregnancies, and details of who requested permission for autopsy. Concordance between diagnoses before death and at autopsy. RESULTS: An autopsy was performed in 209/314 (67%) cases. New information was obtained in 50 (26%) autopsies. In six (3%) cases this information was crucial for future counselling. In 145 (74%) there was complete concordance between the clinical cause of death and the findings at autopsy. From 1994 onwards the autopsy rate in the neonatal unit fell. The only significant factor associated with consent for autopsy was increased gestational age. CONCLUSIONS: Important extra information can be gained at neonatal autopsies. This should help parents to make an informed decision when they are asked to give permission for their baby to have an autopsy. These findings are of particular relevance in view of the recent negative publicity surrounding neonatal autopsies and the general decline in the neonatal autopsy rate over the decade studied. PMID- 11923159 TI - Synergism between allergens and viruses and risk of hospital admission with asthma: case-control study. AB - OBJECTIVE: To investigate the importance of sensitisation and exposure to allergens and viral infection in precipitating acute asthma in adults resulting in admission to hospital. DESIGN: Case-control study. SETTING: Large district general hospital. PARTICIPANTS: 60 patients aged 17-50 admitted to hospital over a year with acute asthma, matched with two controls: patients with stable asthma recruited from the outpatient department and patients admitted to hospital with non-respiratory conditions (inpatient controls). MAIN OUTCOME MEASURES: Atopic status (skin testing and total and specific IgE), presence of common respiratory viruses and atypical bacteria (polymerase chain reaction), dust samples from homes, and exposure to allergens (enzyme linked immunosorbent assay (ELISA): Der p 1, Fel d 1, Can f 1, and Bla g 2). RESULTS: Viruses were detected in 31 of 177 patients. The difference in the frequency of viruses detected between the groups was significant (admitted with asthma 26%, stable asthma 18%, inpatient controls 9%; P=0.04). A significantly higher proportion of patients admitted with asthma (66%) were sensitised and exposed to either mite, cat, or dog allergen than patients with stable asthma (37%) and inpatient controls (15%; P<0.001). Being sensitised and exposed to allergens was an independent associate of the group admitted to hospital (odds ratio 2.3, 95% confidence interval 1.0 to 5.4; P=0.05), whereas the combination of sensitisation, high exposure to one or more allergens, and viral detection considerably increased the risk of being admitted with asthma (8.4, 2.1 to 32.8; P=0.002). CONCLUSIONS: Allergens and viruses may act together to exacerbate asthma. PMID- 11923160 TI - Circadian pattern in occurrence of renal colic in an emergency department: analysis of patients' notes. PMID- 11923162 TI - Autologous transfusion. PMID- 11923161 TI - Randomised controlled trial of short bursts of a potent topical corticosteroid versus prolonged use of a mild preparation for children with mild or moderate atopic eczema. AB - OBJECTIVE: To determine whether a three day burst of a potent corticosteroid is more effective than a mild preparation used for seven days in children with mild or moderate atopic eczema. DESIGN: Randomised, double blind, parallel group study of 18 weeks' duration. SETTING: 13 general practices and a teaching hospital in the Nottingham area. PARTICIPANTS: 174 children with mild or moderate atopic eczema recruited from general practices and 33 from a hospital outpatient clinic. INTERVENTIONS: 0.1% betamethasone valerate applied for three days followed by the base ointment for four days versus 1% hydrocortisone applied for seven days. MAIN OUTCOME MEASURES: Primary outcomes were total number of scratch-free days and number of relapses. Secondary outcomes were median duration of relapses, number of undisturbed nights, disease severity (six area, six sign atopic dermatitis severity scale), scores on two quality of life measures (children's life quality index and dermatitis family impact questionnaire), and number of patients in whom treatment failed in each arm. RESULTS: No differences were found between the two groups. This was consistent for all outcomes. The median number of scratch-free days was 118.0 for the mild group and 117.5 for the potent group (difference 0.5, 95% confidence interval -2.0 to 4.0, P=0.53). The median number of relapses for both groups was 1.0. Both groups showed clinically important improvements in disease severity and quality of life compared with baseline. CONCLUSION: A short burst of a potent topical corticosteroid is just as effective as prolonged use of a milder preparation for controlling mild or moderate atopic eczema in children. PMID- 11923166 TI - Bullying in medicine. Those who can, do; those who can't, bully. PMID- 11923163 TI - ABC of clinical electrocardiography: Broad complex tachycardia-Part II. PMID- 11923167 TI - Violence in the workplace. Delirium should be considered. PMID- 11923168 TI - Language of self harm is somatic and needs to be learnt. PMID- 11923169 TI - Statins as the new aspirin. Conclusions from the heart protection study were premature. PMID- 11923170 TI - Case of cholestatic hepatitis with celecoxib did not fulfil international criteria. PMID- 11923171 TI - Mini-cholecystectomy is often used in gallbladder surgery. PMID- 11923172 TI - Several databases give free access now. PMID- 11923173 TI - Medline and PubMed will be able to synthesise clinical data. PMID- 11923174 TI - Vitamin A programme in Assam probably caused hysteria. PMID- 11923175 TI - Full time forensic pathology service. Practical alternative to forensic pathology service exists. PMID- 11923176 TI - Same shortcomings of NHS have existed for years. PMID- 11923177 TI - Therapeutics needs to be better taught. PMID- 11923182 TI - Proactive and retroactive interference in implicit odor memory. AB - To test the hypothesis that longevity of odor memory is due to strong proactive interference (reduction of new learning by prior learning) and to absence of retroactive interference (reduction of prior memory by new learning), subjects, matched in age and gender with those of a previous experiment, were unknowingly exposed in two sessions to the weak concentrations of lavender or orange used before. Implicit odor memory was later tested in a separate experiment. Comparison of the results with those of the previous experiment showed that both proactive and retroactive interference occurred. These results have implications for the general theory about implicit memory for new associations, which may have to be amended when non-verbal material is used. The longevity of odor memory should be explained by the improbability of occurrence of incidences that provoke retroactive interference rather than by the absence of the retroactive interference itself. PMID- 11923183 TI - A putative pheromone receptor gene is expressed in two distinct olfactory organs in goats. AB - Mammals possess two anatomically and functionally distinct olfactory systems. The olfactory epithelium (OE) detects volatile odorants, while the vomeronasal organ (VNO) detects pheromones that elicit innate reproductive and social behavior within a species. In rodent VNO, three multigene families that encode the putative pheromone receptors, V1Rs, V2Rs and V3Rs, have been expressed. We have identified the V1R homologue genes from goat genomic DNA (gV1R genes). Deduced amino acid sequences of gV1R genes show 40-50% and 20-25% identity to those of rat and mouse V1R and V3R genes, respectively, suggesting that the newly isolated goat receptor genes are members of the V1R gene family. One gene (gV1R1 gene) has an open reading frame that encodes a polypeptide of 309 amino acids. It is expressed not only in VNO but also in OE. In situ hybridization analysis revealed that gV1R1 -expressing cells were localized in neuropithelial layers of VNO and OE. These results suggest that the goat may detect pheromone molecules through two distinct olfactory organs. PMID- 11923184 TI - Adenovirus-mediated WGA gene delivery for transsynaptic labeling of mouse olfactory pathways. AB - Detailed knowledge of neuronal connectivity patterns is indispensable for studies of various aspects of brain functions. We previously established a genetic strategy for visualization of multisynaptic neural pathways by expressing wheat germ agglutinin (WGA) transgene under the control of neuron type-specific promoter elements in transgenic mice and Drosophila. In this paper, we have developed a WGA-expressing recombinant adenoviral vector system and applied it for analysis of the olfactory system. When the WGA-expressing adenovirus was infused into a mouse nostril, various types of cells throughout the olfactory epithelium were infected and expressed WGA protein robustly. WGA transgene products in the olfactory sensory neurons were anterogradely transported along their axons to the olfactory bulb and transsynaptically transferred in glomeruli to dendrites of the second-order neurons, mitral and tufted cells. WGA protein was further conveyed via the lateral olfactory tract to the olfactory cortical areas including the anterior olfactory nucleus, olfactory tubercle, piriform cortex and lateral entorhinal cortex. In addition, transsynaptic retrograde labeling was observed in cholinergic neurons in the horizontal limb of diagonal band, serotonergic neurons in the median raphe nucleus, and noradrenergic neurons in the locus coeruleus, all of which project centrifugal fibers to the olfactory bulb. Thus, the WGA-expressing adenovirus is a useful and powerful tool for tracing neural pathways and could be used in animals that are not amenable to the transgenic technology. PMID- 11923185 TI - Concentration effects of green odor on event-related potential (P300) and pleasantness. AB - The effects of eight compounds, constituting the so-called "natural green odor", including leaf alcohol, on the event-related potential (P300) were investigated. In experiments with a series of single compounds, each of these eight compounds could be characterized by an overall change consisting predominantly of an increase, a decrease or no change in the amplitude of P300, whereas in experiments with a series of two-component mixtures, noticeable synergism could not be demonstrated, contrary to our expectation. Experiments with leaf alcohol (3Z-hexenol) performed at two concentrations showed a significantly different degree of pleasantness and an increase or decrease in the amplitude of P300 depending on their concentration, suggesting that concentration is important in odorant-presentation studies. PMID- 11923186 TI - Spatial representation of odours in the antennal lobe of the moth Spodoptera littoralis (Lepidoptera: Noctuidae). AB - Glomeruli within the antennal lobe (AL) of moths are convergence sites for a large number of olfactory receptor neurons (ORNs). The ORNs target single glomeruli. In the male-specific cluster of glomeruli, the macroglomerular complex (MGC), the input is chemotypic in that each glomerulus of the MGC receives information about a specific component of the conspecific female sex pheromone. Little is known about how neurons that detect other odorants arborize in and amongst glomeruli. The present study focuses on how sex pheromones and biologically relevant semiochemicals are represented in the ALs of both sexes of the moth Spodoptera littoralis. To assess this, we optically measured odour evoked changes of calcium concentration in the ALs. Foci of calcium increase corresponded in size and shape with anatomical glomeruli. More than one glomerulus was normally activated by a specific non-pheromonal odorant and the same glomerulus was activated by several odorants. All odorants and pheromone components tested evoked unique patterns of glomerular activity that were highly reproducible at repeated stimulations within an individual. Odour-evoked patterns were similar between individuals for a given odorant, implicating a spatial olfactory code. In addition, we demonstrated that activity patterns evoked by host-plant related volatiles are similar between males and females. PMID- 11923187 TI - Axon navigation in the mammalian primary olfactory pathway: where to next? AB - The process of establishing long-range neuronal connections can be divided into at least three discrete steps. First, axons need to be stimulated to grow and this growth must be towards appropriate targets. Second, after arriving at their target, axons need to be directed to their topographically appropriate position and in some cases, such as in cortical structures, they must grow radially to reach the correct laminar layer. Third, axons then arborize and form synaptic connections with only a defined subpopulation of potential post-synaptic partners. Attempts to understand these mechanisms in the visual system have been ongoing since pioneer studies in the 1940s highlighted the specificity of neuronal connections in the retino-tectal pathway. These classical systems-based approaches culminated in the 1990s with the discovery that Eph-ephrin repulsive interactions were involved in topographical mapping. In marked contrast, it was the cloning of the odorant receptor family that quickly led to a better understanding of axon targeting in the olfactory system. The last 10 years have seen the olfactory pathway rise in prominence as a model system for axon guidance. Once considered to be experimentally intractable, it is now providing a wealth of information on all aspects of axon guidance and targeting with implications not only for our understanding of these mechanisms in the olfactory system but also in other regions of the nervous system. PMID- 11923188 TI - A computational system for simulating and analyzing arrays of biological and artificial chemical sensors. AB - We have designed an approach for modeling olfactory pathways by which one can explore how the properties of individual receptors affect the information coding capacity of an entire system. The effect of receptor tuning breadth on system performance was explored explicitly. We presented model sensory arrays with sets of stimuli randomly and uniformly distributed in an "olfactory space". Arrays of uniformly sized model receptors responding to 25-35% of the stimuli gave the best performance as measured by the ability to capture the most information about the stimulus set. Arrays of variably sized model receptors that were both more broadly and more narrowly tuned than this optimum could, however, perform better than uniform arrays. This method and the results obtained using it suggest a framework for considering the growing body of evidence on the functional properties of individual olfactory receptor and relay neurons from a systems coding perspective. PMID- 11923189 TI - Amino acid odorants stimulate microvillar sensory neurons. AB - The olfactory epithelium (OE) of zebrafish is populated with ciliated and microvillar olfactory sensory neurons (OSNs). Whether distinct classes of odorants specifically activate either of these unique populations of OSNs is unknown. Previously we demonstrated that zebrafish OSNs could be labeled in an activity-dependent fashion by amino acid but not bile acid odorants. To determine which sensory neuron type was stimulated by amino acid odorants, we labeled OSNs using the ion channel permeant probe agmatine (AGB) and analyzed its distribution with conventional light- and electron-microscope immunocytochemical techniques. Approximately 7% of the sensory epithelium was labeled by AGB exposure alone. Following stimulation with one of the eight amino acids tested, the proportion of labeled epithelium increased from 9% for histidine to 19% for alanine; amino acid stimulated increases in labeling of 2-12% over control labeling. Only histidine failed to stimulate a significant increase in the proportion of labeled OSNs compared to control preparations. Most amino acid sensitive OSNs were located superficially in the epithelium and immuno-electron microscopy demonstrated that the labeled OSNs were predominantly microvillar. Large numbers of nanogold particles (20-60 per 1.5 microm(2)) were associated with microvillar olfactory sensory neurons (MSNs), while few such particles (<15 per 1.5 microm(2)) were observed over ciliated olfactory sensory neurons (CSNs), supporting cells (SCs) and areas without tissue, such as the lumen above the OE. Collectively, these findings indicate that microvillar sensory neurons are capable of detecting amino acid odorants. PMID- 11923190 TI - High-throughput expression profiling techniques. PMID- 11923191 TI - Subtraction-coupled custom microarray analysis for gene discovery and gene expression studies in the CNS. AB - The revolution in our knowledge about the genomes of organisms gives rise to the question, what do we do with this information? The development of techniques allowing high throughput analysis of RNA and protein expression, such as cDNA microarrays, provide for genome-wide analysis of gene expression. These analyses will help bridge the gap between systems and molecular neuroscience. This review discusses the advantages of using a subtractive hybridization technique, such as a representational difference analysis, to generate a custom cDNA microarray enriched for genes relevant to investigating complex, heterogeneous tissues such as those involved in the chemical senses. Real and hypothetical examples of these experiments are discussed. Benefits of this approach over traditional microarray techniques include having a more relevant clone set, the potential for gene discovery and the creation of a new tool to investigate similar systems. Potential pitfalls may include PCR artifacts and the need for sequencing. However, these disadvantages can be overcome so that the coupling of subtraction techniques to microarray screening can be a fruitful approach to a variety of experimental systems. PMID- 11923192 TI - DNA microarray analysis of the aging brain. AB - To examine molecular events associated with brain aging and its retardation by caloric restriction (CR), we have employed high-density oligonucleotide arrays providing data on 6347 genes to define transcriptional patterns in two brain regions (cerebellum and neocortex). Male C57BL/6 mice were either fed normally or subjected to CR. To investigate aging, 5 month (young adult) and 30 month-old normally fed mice were compared. To study CR, 30 month-old control and CR mice were compared. In both brain regions, aging resulted in a gene expression profile suggestive of a marked inflammatory response, oxidative stress and reduced neuronal plasticity and neurotrophic support. In the brain, CR selectively attenuated the age-associated induction of genes encoding inflammatory and stress responses. In addition to providing an improved understanding of the aging process, the use of DNA microarrays generates panels of hundreds of transcriptional biomarkers of molecular aging, providing a new tool to measure biological age on a tissue-specific basis. These studies suggest that genomic approaches may be useful in understanding the molecular basis of the aging process in experimental animals. PMID- 11923193 TI - A transcriptional profile of multicellular development in Dictyostelium discoideum. AB - A distinct feature of development in the simple eukaryote Dictyostelium discoideum is an aggregative transition from a unicellular to a multicellular phase. Using genome-wide transcriptional analysis we show that this transition is accompanied by a dramatic change in the expression of more than 25% of the genes in the genome. We also show that the transcription patterns of these genes are not sensitive to the strain or the nutritional history, indicating that Dictyostelium development is a robust physiological process that is accompanied by stereotypical transcriptional events. Analysis of the two differentiated cell types, spores and stalk cells, and their precursors revealed a large number of differentially expressed genes as well as unexpected patterns of gene expression, which shed new light on the timing and possible mechanisms of cell-type divergence. Our findings provide new perspectives on the complexity of the developmental program and the fraction of the genome that is regulated during development. PMID- 11923194 TI - Comparison of the generic neuronal differentiation and neuron subtype specification functions of mammalian achaete-scute and atonal homologs in cultured neural progenitor cells. AB - In the vertebrate peripheral nervous system, the proneural genes neurogenin 1 and neurogenin 2 (Ngn1 and Ngn2), and Mash1 are required for sensory and autonomic neurogenesis, respectively. In cultures of neural tube-derived, primitive PNS progenitors NGNs promote expression of sensory markers and MASH1 that of autonomic markers. These effects do not simply reflect enhanced neuronal differentiation, suggesting that both bHLH factors also specify neuronal identity like their Drosophila counterparts. At high concentrations of BMP2 or in neural crest stem cells (NCSCs), however, NGNs like MASH1 promote only autonomic marker expression. These data suggest that that the identity specification function of NGNs is more sensitive to context than is that of MASH1. In NCSCs, MASH1 is more sensitive to Notch-mediated inhibition of neurogenesis and cell cycle arrest, than are the NGNs. Thus, the two proneural genes differ in other functional properties besides the neuron subtype identities they can promote. These properties may explain cellular differences between MASH1- and NGN-dependent lineages in the timing of neuronal differentiation and cell cycle exit. PMID- 11923196 TI - Overexpression of Snail family members highlights their ability to promote chick neural crest formation. AB - The Snail gene family of transcription factors plays crucial roles in different morphogenetic processes during the development of vertebrate and invertebrate embryos. In previous studies of function interference for one of the family members, Slug, we showed its involvement and neural crest formation in the chick embryo. Now we have carried out a series of gain-of-function experiments in which we show that Slug overexpression in the neural tube of the chick embryo induces an increase in neural crest production. The analysis of electroporated embryos shows that Slug can induce the expression of rhoB and an increase in the number of HNK-1-positive migratory cells, indicating that it lies upstream of them in the genetic cascade of neural crest development. The increase in neural crest production after Slug overexpression was confined to the cranial region, indicating that the mechanisms of crest induction somehow differ between head and trunk. The expression of the two vertebrate family members, Slug and Snail, is peculiar with respect to the neural crest. Slug is not expressed in the premigratory crest in the mouse, whereas it is expressed in this cell population in the chick and the opposite is true for Snail ( Sefton, M., Sanchez, S. and Nieto M. A. (1998) Development 125, 3111-3121). This raises the question of whether they can be functionally equivalent. To test this hypothesis both intra- and interspecies, we have performed a series of ectopic expression experiments by electroporating chick and mouse Snail in the chick embryo hindbrain. We observe that both genes elicit the same responses in the neural tube. Our results indicate that they can be functionally equivalent, although the embryos show a higher response to the endogenous gene, chick Slug. PMID- 11923195 TI - HUA ENHANCER2, a putative DExH-box RNA helicase, maintains homeotic B and C gene expression in Arabidopsis. AB - Reproductive organ identity in Arabidopsis is controlled by the B, C and SEPALLATA classes of floral homeotic genes. We have identified a recessive mutation in a novel gene, HUA ENHANCER2, which, when combined with mutations in two weak class C genes, HUA1 and HUA2, leads to the production of third whorl sepal-petal-stamens and fourth whorl sepal-carpels. Quadruple mutant analysis and in situ localization of A, B, C and SEPALLATA floral homeotic RNAs suggest that HUA ENHANCER2 is required for the maintenance of B and C gene expression in the reproductive whorls. In addition to its role in floral homeotic gene expression, HUA ENHANCER2 is required for normal spacing and number of perianth organ primordia. We show that HUA ENHANCER2 encodes a putative DExH-box RNA helicase that is expressed in specific patterns in the inflorescence meristem and developing flowers. As a possible ortholog of the yeast exosome-associated protein, Dob1p (Mtr4p), HUA ENHANCER2 may affect floral organ spacing and identity through the regulation of protein synthesis or mRNA degradation. Therefore, our studies on HUA ENHANCER2 not only demonstrate that B and C gene expression is established and maintained separately, but also implicate the existence of post-transcriptional mechanisms in the maintenance of B and C gene expression. PMID- 11923197 TI - A pea seed mutant affected in the differentiation of the embryonic epidermis is impaired in embryo growth and seed maturation. AB - During legume seed development the epidermis of the embryos differentiates into a transfer cell layer which mediates nutrient uptake during the storage phase. This specific function of the epidermal cells is acquired at the onset of embryo maturation. We investigated this process in the pea seed mutant E2748. The epidermal cells of the mutant embryo, instead of turning into transfer cells, enlarge considerably and become vacuolated and tightly associated with adjacent seed tissues. Expression of a sucrose transporter gene that is upregulated in wild-type transfer cells decreases in the mutant and changes its spatial pattern. This indicates that the outermost cell layer of mutant cotyledons cannot acquire transfer cell morphology but loses epidermal cell identity and does not function as a sucrose uptake system. Seed coat growth as well as composition, concentration and dynamics of sugars within the endospermal vacuole are unchanged. The loss of epidermal identity has severe consequences for further embryo development and is followed by disruption of the symplast within the parenchyma, the breach of the developmental gradient, lower sucrose and starch levels and initiation of callus-like growth. It is concluded that the E2748 gene controls differentiation of the cotyledonary epidermis into transfer cells and thus is required for the regional specialisation with a function in embryo nutrition. PMID- 11923198 TI - The homeoprotein Xiro1 is required for midbrain-hindbrain boundary formation. AB - The isthmic organizer, which patterns the anterior hindbrain and midbrain, is one of the most studied secondary organizers. In recent years, new insights have been reported on the molecular nature of its morphogenetic activity. Studies in chick, mouse and zebrafish have converged to show that mutually repressive interactions between the homeoproteins encoded by Otx and Gbx genes position this organizer in the neural primordia. We present evidence that equivalent, in addition to novel, interactions between these and other genes operate in Xenopus embryos to position the isthmic organizer. We made use of fusion proteins in which we combined Otx2 or Gbx2 homeodomains with the E1A activation domain or the EnR repressor element which were then injected into embryos. Our results show that Otx2 and Gbx2 are likely to be transcriptional repressors, and that these two proteins repress each other transcription. Our experiments show that the interaction between these two proteins is required for the positioning of the isthmic organizer genes Fgf8, Pax2 and En2. In this study we also developed a novel in vitro assay for the study of the formation of this organizer. We show that conjugating animal caps previously injected with Otx2 and Gbx2 mRNAs recreate the interactions required for the induction of the isthmic organizer. We have used this assay to determine which cells produce and which cells receive the Fgf signal. Finally, we have added a novel genetic element to this process, Xiro1, which encode another homeoprotein. We show that the Xiro1 expression domain overlaps with territories expressing Otx2, Gbx2 and Fgf8. By expressing wild-type or dominant negative forms of Xiro1, we show that this gene activates the expression of Gbx2 in the hindbrain. In addition, Xiro1 is required in the Otx2 territory to allow cells within this region to respond to the signals produced by adjacent Gbx2 cells. Moreover, Xiro1 is absolutely required for Fgf8 expression at the isthmic organizer. We discuss a model where Xiro1 plays different roles in regulating the genetic cascade of interactions between Otx2 and Gbx2 that are necessary for the specification of the isthmic organizer. PMID- 11923199 TI - The elongation factors Pandora/Spt6 and Foggy/Spt5 promote transcription in the zebrafish embryo. AB - Precise temporal and spatial control of transcription is a fundamental component of embryonic development. Regulation of transcription elongation can act as a rate-limiting step during mRNA synthesis. The mechanisms of stimulation and repression of transcription elongation during development are not yet understood. We have identified a class of zebrafish mutations (pandora, sk8 and s30) that cause multiple developmental defects, including discrete problems with pigmentation, tail outgrowth, ear formation and cardiac differentiation. We demonstrate that the pandora gene encodes a protein similar to Spt6, a proposed transcription elongation factor. Additionally, the sk8 and s30 mutations are null alleles of the foggy/spt5 locus, which encodes another transcription elongation factor. Through real-time RT-PCR analysis, we demonstrate that Spt6 and Spt5 are both required for efficient kinetics of hsp70 transcription in vivo. Altogether, our results suggest that Spt6 and Spt5 play essential roles of comparable importance for promoting transcription during embryogenesis. This study provides the first genetic evidence for parallel functions of Spt6 and Spt5 in metazoans and establishes a system for the future analysis of transcription elongation during development. PMID- 11923200 TI - Emx2 regulates the proliferation of stem cells of the adult mammalian central nervous system. AB - The appropriate control of proliferation of neural precursors has fundamental implications for the development of the central nervous system and for cell homeostasis/replacement within specific brain regions throughout adulthood. The role of genetic determinants in this process is largely unknown. We report the expression of the homeobox transcription factor Emx2 within the periventricular region of the adult telencephalon. This neurogenetic area displays a large number of multipotent stem cells. Adult neural stem cells isolated from this region do express Emx2 and down-regulate it significantly upon differentiation into neurons and glia. Abolishing or, increasing Emx2 expression in adult neural stem cells greatly enhances or reduces their rate of proliferation, respectively. We determined that altering the expression of Emx2 affects neither the cell cycle length of adult neural stem cells nor their ability to generate neurons and glia. Rather, when Emx2 expression is abolished, the frequency of symmetric divisions that generate two stem cells increases, whereas it decreases when Emx2 expression is enhanced. PMID- 11923201 TI - spiel ohne grenzen/pou2 is required for zebrafish hindbrain segmentation. AB - Segmentation of the vertebrate hindbrain leads to the formation of a series of rhombomeres with distinct identities. In mouse, Krox20 and kreisler play important roles in specifying distinct rhombomeres and in controlling segmental identity by directly regulating rhombomere-specific expression of Hox genes. We show that spiel ohne grenzen (spg) zebrafish mutants develop rhombomeric territories that are abnormal in both size and shape. Rhombomere boundaries are malpositioned or absent and the segmental pattern of neuronal differentiation is perturbed. Segment-specific expression of hoxa2, hoxb2 and hoxb3 is severely affected during initial stages of hindbrain development in spg mutants and the establishment of krx20 (Krox20 ortholog) and valentino (val; kreisler ortholog) expression is impaired. spg mutants carry loss-of-function mutations in the pou2 gene. pou2 is expressed at high levels in the hindbrain primordium of wild-type embryos prior to activation of krx20 and val. Widespread overexpression of Pou2 can rescue the segmental krx20 and val domains in spg mutants, but does not induce ectopic expression of these genes. This suggests that spg/pou2 acts in a permissive manner and is essential for normal expression of krx20 and val. We propose that spg/pou2 is an essential component of the regulatory cascade controlling hindbrain segmentation and acts before krx20 and val in the establishment of rhombomere precursor territories. PMID- 11923202 TI - Disruption of mesodermal enhancers for Igf2 in the minute mutant. AB - The radiation-induced mutation minute (Mnt) in the mouse leads to intrauterine growth retardation with paternal transmission and has been linked to the distal chromosome 7 cluster of imprinted genes. We show that the mutation is an inversion, whose breakpoint distal to H19 disrupts and thus identifies an enhancer for Igf2 expression in skeletal muscle and tongue, and separates the gene from other mesodermal and extra-embryonic enhancers. Paternal transmission of Mnt leads to drastic downregulation of Igf2 transcripts in all mesodermal tissues and the placenta. Maternal transmission leads to methylation of the H19 differentially methylated region (DMR) and silencing of H19, showing that elements 3' of H19 can modify the maternal imprint. Methylation of the maternal DMR leads to biallelic expression of Igf2 in endodermal tissues and foetal overgrowth, demonstrating that methylation in vivo can open the chromatin boundary upstream of H19. Our work shows that most known enhancers for Igf2 are located 3' of H19 and establishes an important genetic paradigm for the inheritance of complex regulatory mutations in imprinted gene clusters. PMID- 11923204 TI - Onset of neuronal differentiation is regulated by paraxial mesoderm and requires attenuation of FGF signalling. AB - While many neuronal differentiation genes have been identified, we know little about what determines when and where neurons will form and how this process is coordinated with the differentiation of neighbouring tissues. In most vertebrates the onset of neuronal differentiation takes place in the spinal cord in a head to tail sequence. Here we demonstrate that the changing signalling properties of the adjacent paraxial mesoderm control the progression of neurogenesis in the chick spinal cord. We find an inverse relationship between the expression of caudal neural genes in the prospective spinal cord, which is maintained by underlying presomitic mesoderm and FGF signalling, and neuronal differentiation, which is repressed by such signals and accelerated by somitic mesoderm. We show that key to this interaction is the ability of somitic mesoderm to repress Fgf8 transcription in the prospective spinal cord. Our findings further indicate that attenuation of FGF signalling in the prospective spinal cord is a prerequisite for the onset of neuronal differentiation and may also help to resolve mesodermal and neural cell fates. However, inhibition of FGF signalling alone does not promote the formation of neurons, which requires still further somite signalling. We propose a model in which signalling from somitic tissue promotes the differentiation of the spinal cord and serves to co-ordinate neural and mesodermal development. PMID- 11923203 TI - The activity of the Drosophila morphogenetic protein Bicoid is inhibited by a domain located outside its homeodomain. AB - The Drosophila morphogenetic protein Bicoid (Bcd) is a homeodomain-containing activator that stimulates the expression of target genes during early embryonic development. We demonstrate that a small domain of Bcd located immediately N terminally of the homeodomain represses its own activity in Drosophila cells. This domain, referred to as a self-inhibitory domain, works as an independent module that does not rely on any other sequences of Bcd and can repress the activity of heterologous activators. We further show that this domain of Bcd does not affect its properties of DNA binding or subcellular distribution. A Bcd derivative with point mutations in the self-inhibitory domain severely affects pattern formation and target gene expression in Drosophila embryos. We also provide evidence to suggest that the action of the self-inhibitory domain requires a Drosophila co-factor(s), other than CtBP or dSAP18. Our results suggest that proper action of Bcd as a transcriptional activator and molecular morphogen during embryonic development is dependent on the downregulation of its own activity through an interaction with a novel co-repressor(s) or complex(es). PMID- 11923205 TI - Annexin IV (Xanx-4) has a functional role in the formation of pronephric tubules. AB - Vertebrate kidney organogenesis is characterised by the successive formation of the pronephros, the mesonephros and the metanephros. The pronephros is the first to form and is the functional embryonic kidney of lower vertebrates; although it is vestigial in higher vertebrates, it is a necessary precursor for the other kidney types. The Xenopus pronephros is a simple paired organ; each nephron consists of a single large glomus, one set of tubules and a single duct. The simple organisation of the pronephros and the amenability of Xenopus laevis embryos to manipulation make the Xenopus pronephros an attractive system in which to study organogenesis. It has been shown that pronephric tubules can be induced to form in presumptive ectodermal tissue by treatment with RA and activin. We have used this system in a subtractive hybridisation screen that resulted in the cloning of Xenopus laevis annexin IV (Xanx-4). Xanx-4 transcripts are specifically located to the developing pronephric tubules, and the protein to the luminal surface of these tubules. Temporal expression shows zygotic transcription is upregulated at the time of pronephric tubule specification and persists throughout pronephric development. The temporal and spatial expression pattern of Xanx-4 suggests it may have a role in pronephric tubule development. Overexpression of Xanx-4 yields no apparent phenotype, but Xanx-4 depletion, using morpholinos, produces a shortened, enlarged tubule phenotype. The phenotype observed can be rescued by co-injection of Xanx-4 mRNA. Although the function of annexins is not yet clear, studies have suggested a role for annexins in a number of cellular processes. Annexin IV has been shown to have an inhibitory role in the regulation of epithelial calcium-activated chloride ion conductance. The enlarged pronephric tubule phenotype observed may be attributed to incorrect modulation of exocytosis, membrane plasticity or ion channels and/or water homeostasis. In this study, we demonstrate an in vivo role for annexin IV in the development of the pronephric tubules in Xenopus laevis. PMID- 11923207 TI - The MAPK pathway triggers activation of Nek2 during chromosome condensation in mouse spermatocytes. AB - Chromosome condensation during the G2/M progression of mouse pachytene spermatocytes induced by the phosphatase inhibitor okadaic acid (OA) requires the activation of the MAPK Erk1. In many cell systems, p90Rsks are the main effectors of Erk1/2 function. We have identified p90Rsk2 as the isoform that is specifically expressed in mouse spermatocytes and have shown that it is activated during the OA-triggered meiotic G2/M progression. By using the MEK inhibitor U0126, we have demonstrated that activation of p90Rsk2 during meiotic progression requires activation of the MAPK pathway. Immunofluorescence analysis indicates that activated Erks and p90Rsk2 are tightly associated with condensed chromosomes during the G2/M transition in meiotic cells. We also found that active p90Rsk2 was able to phosphorylate histone H3 at Ser10 in vitro, but that the activation of the Erk1/p90Rsk2 pathway was not necessary for phosphorylation of H3 in vivo. Furthermore, phosphorylation of H3 was not sufficient to cause condensation of meiotic chromosomes in mouse spermatocytes. Other proteins known to associate with chromatin may represent effectors of Erk1 and p90Rsk2 during chromosome condensation. Nek2 (NIMA-related kinase 2), which associates with chromosomes, plays an active role in chromatin condensation and is stimulated by treatment of pachytene spermatocytes with okadaic acid. We show that inhibition of the MAPK pathway by preincubation of spermatocytes with U0126 suppresses Nek2 activation, and that incubation of spermatocyte cell extracts with activated p90Rsk2 causes stimulation of Nek2 kinase activity. Furthermore, we show that the Nek2 kinase domain is a substrate for p90Rsk2 phosphorylation in vitro. These data establish a connection between the Erk1/p90Rsk2 pathway, Nek2 activation and chromosome condensation during the G2/M transition of the first meiotic prophase. PMID- 11923206 TI - Inhibition of Rho family GTPases by Rho GDP dissociation inhibitor disrupts cardiac morphogenesis and inhibits cardiomyocyte proliferation. AB - Studies of Rho GTPases in Drosophila and Xenopus suggest that Rho family proteins may play an important role in embryogenesis. A reverse genetic approach was employed to explore the role of Rho GTPases in murine cardiac development. Cardiac-specific inhibition of Rho family protein activities was achieved by expressing Rho GDIalpha, a specific GDP dissociation inhibitor for Rho family proteins, using the alpha-myosin heavy chain promoter, active at embryonic day (E)8.0 during morphogenesis of the linear heart tube. RhoA, Rac1 and Cdc42 activities were significantly inhibited, as shown by decreased membrane translocation of these proteins in the transgenic hearts. Transgenic F1 mice for each of two independent lines expressing the highest levels of the transgene, died around E10.5. Homozygotes of the middle copy-number lines, in which Rho GDIalpha expression was increased four-fold over normal levels, were also embryonic lethal. Cardiac morphogenesis in these embryos was disrupted, with incomplete looping, lack of chamber demarcation, hypocellularity and lack of trabeculation. Cell proliferation was inhibited in the transgenic hearts, as shown by immunostaining with anti-phosphohistone H3, a marker of mitosis. In addition, ventricular hypoplasia was associated with up-regulation of p21, an inhibitor of cyclin-dependent kinases, and with down-regulation of cyclin A, while cell survival was not affected. These results reveal new biological functions for Rho family proteins as essential determinants of cell proliferation signals at looping and chamber maturation stages in mammalian cardiac development. PMID- 11923209 TI - Spatial patterns of ecdysteroid receptor activation during the onset of Drosophila metamorphosis. AB - Ecdysteroid signaling in insects is transduced by a heterodimer of the EcR and USP nuclear receptors. In order to monitor the temporal and spatial patterns of ecdysteroid signaling in vivo we established transgenic animals that express a fusion of the GAL4 DNA binding domain and the ligand binding domain (LBD) of EcR or USP, combined with a GAL4-dependent lacZ reporter gene. The patterns of beta galactosidase expression in these animals indicate where and when the GAL4-LBD fusion protein has been activated by its ligand in vivo. We show that the patterns of GAL4-EcR and GAL4-USP activation at the onset of metamorphosis reflect what would be predicted for ecdysteroid activation of the EcR/USP heterodimer. No activation is seen in mid-third instar larvae when the ecdysteroid titer is low, and strong widespread activation is observed at the end of the instar when the ecdysteroid titer is high. In addition, both GAL4-EcR and GAL4-USP are activated in larval organs cultured with 20-hydroxyecdysone (20E), consistent with EcR/USP acting as a 20E receptor. We also show that GAL4-USP activation depends on EcR, suggesting that USP requires its heterodimer partner to function as an activator in vivo. Interestingly, we observe no GAL4-LBD activation in the imaginal discs and ring glands of late third instar larvae. Addition of 20E to cultured mid-third instar imaginal discs results in GAL4-USP activation, but this response is not seen in imaginal discs cultured from late third instar larvae, suggesting that EcR/USP loses its ability to function as an efficient activator in this tissue. We conclude that EcR/USP activation by the systemic ecdysteroid signal may be spatially restricted in vivo. Finally, we show that GAL4-EcR functions as a potent and specific dominant negative at the onset of metamorphosis, providing a new tool for characterizing ecdysteroid signaling pathways during development. PMID- 11923208 TI - Early embryonic expression of FGF4/6/9 gene and its role in the induction of mesenchyme and notochord in Ciona savignyi embryos. AB - In early Ciona savignyi embryos, nuclear localization of beta-catenin is the first step of endodermal cell specification, and triggers the activation of various target genes. A cDNA for Cs-FGF4/6/9, a gene activated downstream of beta catenin signaling, was isolated and shown to encode an FGF protein with features of both FGF4/6 and FGF9/20. The early embryonic expression of Cs-FGF4/6/9 was transient and the transcript was seen in endodermal cells at the 16- and 32-cell stages, in notochord and muscle cells at the 64-cell stage, and in nerve cord and muscle cells at the 110-cell stage; the gene was then expressed again in cells of the nervous system after neurulation. When the gene function was suppressed with a specific antisense morpholino oligo, the differentiation of mesenchyme cells was completely blocked, and the fate of presumptive mesenchyme cells appeared to change into that of muscle cells. The inhibition of mesenchyme differentiation was abrogated by coinjection of the morpholino oligo and synthetic Cs-FGF4/6/9 mRNA. Downregulation of beta-catenin nuclear localization resulted in the absence of mesenchyme cell differentiation due to failure of the formation of signal producing endodermal cells. Injection of synthetic Cs-FGF4/6/9 mRNA in beta catenin-downregulated embryos evoked mesenchyme cell differentiation. These results strongly suggest that Cs-FGF4/6/9 produced by endodermal cells acts an inductive signal for the differentiation of mesenchyme cells. On the other hand, the role of Cs-FGF4/6/9 in the induction of notochord cells is partial; the initial process of the induction was inhibited by Cs-FGF4/6/9 morpholino oligo, but notochord-specific genes were expressed later to form a partial notochord. PMID- 11923210 TI - Drosophila Apc1 and Apc2 regulate Wingless transduction throughout development. AB - Inactivation of the Adenomatous Polyposis Coli (APC) tumor suppressor triggers the development of most colorectal carcinomas. APC is required for targeted degradation of beta-catenin, the central transcriptional activator in the Wnt/Wingless (Wg) signal transduction pathway; however, the precise biochemical functions of APC remain uncertain. The two Drosophila homologs of APC (Apc1 and Apc2) appear to have predominantly different tissue distributions, different subcellular localizations and mutually exclusive phenotypes upon inactivation. Unexpectedly, we have found that despite these differences, simultaneous reduction in both Drosophila Apc proteins results in the global nuclear accumulation of beta-catenin and the constitutive activation of Wg transduction throughout development. This redundancy extends even to functions previously thought to be specific to the individual Apc homologs. Together, these results reveal that the combined activity of Apc1 and Apc2 allows a tight regulation of transcriptional activation by beta-catenin and suggest that APC proteins are required for the regulation of Wnt transduction in all cells. PMID- 11923212 TI - Permeability barrier dysfunction in transgenic mice overexpressing claudin 6. AB - A defective epidermal permeability barrier (EPB) in premature birth remains a leading cause of neonatal death as a result of its associated complications, which include poor temperature stability, infection by micro-organisms through the skin, and the outflow of water. Despite its importance in survival, the mechanisms involved in the formation and maintenance of the EPB are not well understood. To address the possibility that claudins, a new superfamily of tight junctional molecules, are involved, we engineered transgenic mice with claudin 6 (Cldn6) overexpressed via the involucrin (Inv) promoter. Interestingly, the Inv Cldn6 transgenic animals die within 2 days of birth, apparently due to the lack of an intact EPB as evidenced by increased water loss and the penetration of X gal through the skin. Barrier dysfunction was manifested biochemically by the aberrant expression of late epidermal differentiation markers, including K1, filaggrin, loricrin, transglutaminase 3, involucrin, repetin, members of the SPRR family and the transcriptional regulator Klf4. The overall claudin profile of the epidermis was also modified. Our data suggest that repetin and SPRR1A and 2A are downregulated in response to the downregulation of Klf4 in the transgenic animals, which would contribute to decreased protein crossbridging leading to fragile, defective cornified envelopes. These results provide new insights into the role of claudin 6 in epithelial differentiation and EPB formation. In addition, the epidermal phenotype of these transgenic mice, which is very reminiscent of that in pre-term infant skin, suggest that they will be an important and novel model for studies on human premature EPB-related morbidity. PMID- 11923211 TI - PAG-3, a Zn-finger transcription factor, determines neuroblast fate in C. elegans. AB - During Caenorhabditis elegans development, the patterns of cell divisions, cell fates and programmed cell deaths are reproducible from animal to animal. In a search for mutants with abnormal patterns of programmed cell deaths in the ventral nerve cord, we identified mutations in the gene pag-3, which encodes a zinc-finger transcription factor similar to the mammalian Gfi-1 and Drosophila Senseless proteins. In pag-3 mutants, specific neuroblasts express the pattern of divisions normally associated with their mother cells, producing with each reiteration an abnormal anterior daughter neuroblast and an extra posterior daughter cell that either terminally differentiates or undergoes programmed cell death, which accounts for the extra cell corpses seen in pag-3 mutants. In addition, some neurons do not adopt their normal fates in pag-3 mutants. The phenotype of pag-3 mutants and the expression pattern of the PAG-3 protein suggest that in some lineages pag-3 couples the determination of neuroblast cell fate to subsequent neuronal differentiation. We propose that pag-3 counterparts in other organisms determine blast cell identity and for this reason may lead to cell lineage defects and cell proliferation when mutated. PMID- 11923213 TI - The Snail repressor positions Notch signaling in the Drosophila embryo. AB - The maternal Dorsal nuclear gradient initiates the differentiation of the mesoderm, neurogenic ectoderm and dorsal ectoderm in the precellular Drosophila embryo. Each tissue is subsequently subdivided into multiple cell types during gastrulation. We have investigated the formation of the mesectoderm within the ventral-most region of the neurogenic ectoderm. Previous studies suggest that the Dorsal gradient works in concert with Notch signaling to specify the mesectoderm through the activation of the regulatory gene sim within single lines of cells that straddle the presumptive mesoderm. This model was confirmed by misexpressing a constitutively activated form of the Notch receptor, Notch(IC), in transgenic embryos using the eve stripe2 enhancer. The Notch(IC) stripe induces ectopic expression of sim in the neurogenic ectoderm where there are low levels of the Dorsal gradient. sim is not activated in the ventral mesoderm, due to inhibition by the localized zinc-finger Snail repressor, which is selectively expressed in the ventral mesoderm. Additional studies suggest that the Snail repressor can also stimulate Notch signaling. A stripe2-snail transgene appears to induce Notch signaling in 'naive' embryos that contain low uniform levels of Dorsal. We suggest that these dual activities of Snail, repression of Notch target genes and stimulation of Notch signaling, help define precise lines of sim expression within the neurogenic ectoderm. PMID- 11923214 TI - Axial skeletal defects caused by mutation in the spondylocostal dysplasia/pudgy gene Dll3 are associated with disruption of the segmentation clock within the presomitic mesoderm. AB - A loss-of-function mutation in the mouse delta-like3 (Dll3) gene has been generated following gene targeting, and results in severe axial skeletal defects. These defects, which consist of highly disorganised vertebrae and costal defects, are similar to those associated with the Dll3-dependent pudgy mutant in mouse and with spondylocostal dysplasia (MIM 277300) in humans. This study demonstrates that Dll3(neo) and Dll3(pu) are functionally equivalent alleles with respect to the skeletal dysplasia, and we suggest that the three human DLL3 mutations associated with spondylocostal dysplasia are also functionally equivalent to the Dll3(neo) null allele. Our phenotypic analysis of Dll3(neo)/Dll3(neo) mutants shows that the developmental origins of the skeletal defects lie in delayed and irregular somite formation, which results in the perturbation of anteroposterior somite polarity. As the expression of Lfng, Hes1, Hes5 and Hey1 is disrupted in the presomitic mesoderm, we suggest that the somitic aberrations are founded in the disruption of the segmentation clock that intrinsically oscillates within presomitic mesoderm. PMID- 11923215 TI - Elimination of vascular fibrointimal hyperplasia by somatostatin receptor 1,4 selective agonist. AB - The somatostatin analogs octreotide and lanreotide, selective to receptor subtypes 2 and 5, failed clinical efficacy for the prevention of restenosis after percutaneous transluminal angioplasty. These findings might have been the result of targeting a wrong subset of receptors. In rat arteries, subtypes 1 and 4 are expressed 3-4 times more prominently than 2 and 5, and subtype 1 is the nearly exclusive subtype in atherosclerotic human vessels. Here, we demonstrate that daily s.c. injections (50-500 microg/kg/d) of CH275 (DesAA1,2,5(D W8,IAmp9)Somatostatine-14), selective to subtypes 1 and 4, dose-dependently inhibited intimal hyperplasia 14 days after rat carotid denudation injury (for intimal area P=0.0002 across the dose range). CH275 was more effective than somatostatin-14 (equal affinity to all five subtypes, P=0.03), or octreotide (selective to subtypes 2 and 5, P=0.098). When rats were given the peptides for 14 days with end-point at 28 days, CH275 still significantly inhibited intimal area expansion. Both CH275 and octreotide inhibited the outgrowth of cells from postinjury aortic tissue punch-explants and the distance migrated in vitro, but not cell replication, which indicated that the effects of somatostatin analogs were directed on the migration of intimal cell progenitors rather than on their proliferation. PMID- 11923216 TI - Apoptosis in hypoxic human pancreatic islets correlates with HIF-1alpha expression. AB - To become insulin independent, patients with type 1 diabetes mellitus require transplantation of at least two donor pancreata because of massive beta-cell loss in the early post-transplantation period. Many studies describing the introduction of new immunosuppressive protocols have shown that this loss is due to not only immunological events but also nonimmunological factors. To test to what extent hypoxia may contribute to early graft loss, we analyzed the occurrence of apoptotic events and the expression of hypoxia-inducible factor 1 (HIF-1), a heterodimeric transcription factor consisting of an oxygen-dependent alpha subunit and a constitutive beta subunit. Histological analysis of human and rat islets revealed nuclear pyknosis as early as 6 h after hypoxic exposure (1% O2). Moreover, immunoreactivity to activated caspase-3 was observed in the core region of isolated human islets. Of note, both of these markers of apoptosis topographically overlap with HIF-1alpha immunoreactivity. HIF-1alpha mRNA was detected in islets from human and rat as well as in several murine beta-cell lines. When exposed to hypoxia, mouse insulinoma cells (MIN6) had an increased HIF-1alpha protein level, whereas its mRNA level did not alter. In conclusion, our data provide convincing evidence that reduced oxygenation is an important cause of beta-cell loss and suggest that HIF-1alpha protein level is an indicator for hypoxic regions undergoing apoptotic cell death. These observations suggest that gene expression under the control of HIF-1 represents a potential therapeutic tool for improving engraftment of transplanted islets. PMID- 11923217 TI - Specific PAF antagonist WEB-2086 induces terminal differentiation of murine and human leukemia cells. AB - A pharmacological approach to neoplasia by differentiation therapy relies on the availability of cytodifferentiating agents whose antitumor efficacy is usually assayed first on malignant cells in vitro. Using murine erythroleukemia cells (MELCs) as the model, we found that WEB-2086, a triazolobenzodiazepine-derived PAF antagonist originally developed as an anti-inflammatory drug, induces a dose dependent inhibition of MELC growth and hemoglobin accumulation as a result of a true commitment to differentiation. MELCs treated for 5 days with 1 mM WEB-2086 show greater than or equal to 85% benzidine-positive cells, increased expression of alpha- and beta-globin genes, and down-regulation of c-Myb. This differentiation pattern, which does not involve histone H4 acetylation and is abrogated by the action of phorbol 12-myristate 13-acetate, recalls the pattern induced by hexamethylene bisacetamide (HMBA). In addition to MELCs, human erythroleukemia K562 and HEL and myeloid HL60 cells are massively committed to maturation by WEB-2086 and, with some differences, by its analog, WEB-2170. This suggests that WEB-2086, structurally distant from other known inducers, might be a member of a new class of cytodifferentiation agents active on a broad range of transformed cells in vitro and useful, prospectively, for anticancer therapy due to their high tolerability in vivo. PMID- 11923218 TI - Recovery from osteoporosis through skeletal growth: early bone mass acquisition has little effect on adult bone density. AB - It is often assumed that bone mineral accretion should be optimized throughout childhood to maximize peak bone mass. In contrast, we hypothesized that bone mineral acquisition early in life would have little or no effect on adult bone mass because many areas of the juvenile skeleton are replaced in toto through skeletal growth. To test this hypothesis, we induced osteoporosis by administering dexamethasone to 5-week-old rabbits for 5 weeks and then allowed them to recover for 16 weeks. Tibial bone mineral density (ash weight/volume) was decreased in the dexamethasone-treated animals at the end of treatment but recovered completely. Bone structure in the femur was assessed by histomorphometry. Trabecular and cortical bone in the distal metaphysis was made osteoporotic by dexamethasone, but was then replaced through endochondral bone formation and recovered. Periosteal bone formation rate in the diaphysis was decreased during dexamethasone treatment but afterwards rebounded above controls and normalized cortical width. Our data suggest that bone mineral acquisition early in life has little effect on adult bone density because the juvenile bone is largely replaced through growth. If this concept generalizes, then interventions to maximize peak bone mass should be directed at adolescents rather than young children. PMID- 11923219 TI - A novel secreted splice variant of vascular endothelial cell growth inhibitor. AB - Vascular endothelial cell growth inhibitor (VEGI), a member of the tumor necrosis factor (TNF) family, is an endothelial cell-specific inhibitor of angiogenesis. Overexpression by cancer cells of a secretable VEGI fusion protein resulted in abrogation of xenograft tumor progression, but overexpression of full-length VEGI was completely without effect. This finding indicates that secretion is essential for VEGI action. Here we report the identification of two new VEGI isoforms consisting of 251 and 192 amino acid residues. Both isoforms show endothelial cell-specific expression and share a C-terminal 151-residue segment with the previously described VEGI, which comprises 174 residues. The isoforms are generated from a 17 kb human gene by alternative splicing. Their expression is regulated in parallel by inflammatory cytokines TNF-alpha and interferon-gamma. VEGI-251, the most abundant isoform, contains a putative secretion signal. VEGI protein is detected in conditioned media of endothelial cells and VEGI-251 transfected mammalian cells. Overexpression of VEGI-251 in endothelial cells causes dose-dependent cell death. VEGI-251-transfected cancer cells form xenograft tumors of reduced growth rate and microvessel density compared with tumors of empty vector transfectants. These findings support the view that endothelial cell-secreted VEGI may function as an autocrine inhibitor of angiogenesis and a naturally existing modulator of vascular homeostasis. PMID- 11923220 TI - Cell migration through defined, synthetic ECM analogs. AB - We have developed synthetic hydrogel extracellular matrix (ECM) analogues that can be used to study mechanisms involved in cell migration, such as receptor ligand interactions and proteolysis. The biomimetic hydrogels consist of bioinert polyethylene glycol diacrylate derivatives with proteolytically degradable peptide sequences included in the backbone of the polymer and adhesive peptide sequences grafted to the network. Hydrogels have been developed that degrade as cells secrete proteolytic enzymes. Adhesive peptide sequences grafted to the hydrogel provide ligands that can interact with receptors on the cell surface to mediate adhesion and spreading. In this study, we have characterized the effects of adhesive ligand density on fibroblast migration through collagenase-degradable and plasmin-degradable hydrogels and on smooth muscle cell migration through elastase-degradable hydrogels. In all three cases, we found that cell migration has a biphasic dependence on adhesion ligand concentration, with optimal migration at intermediate ligand levels. Furthermore, both adhesive and proteolytically degradable sequences were required for cell migration to occur. These synthetic ECM analogues may be useful for 3-D mechanistic studies of many aspects of cell migration PMID- 11923221 TI - Phytanic acid, a natural peroxisome proliferator-activated receptor (PPAR) agonist, regulates glucose metabolism in rat primary hepatocytes. AB - Phytanic acid, a metabolite of the chlorophyll molecule, is part of the human diet and is present in normal human serum at low micromolar concentrations. It was previously shown to be a ligand of the 9-cis-retinoic acid receptor and peroxisome proliferator-activated receptor (PPAR) a. PPAR agonists are widely used in the treatment of type 2 diabetes. Here, we report that phytanic acid is not only a transactivator of PPARa, but it also acts via PPARb and PPARg in CV-1 cells that have been cotransfected with the respective full-length receptor and an acyl-CoA oxidase-PPAR-responsive element-luciferase construct. We observed that, in contrast to other fatty acids, phytanic acid at physiological concentrations enhances uptake of 2-deoxy-D-glucose in rat primary hepatocytes. This result could be explained by the increase in mRNA expression of glucose transporters-1 and -2 and glucokinase, as determined by quantitative real-time reverse transcriptase-polymerase chain reaction. Compared with the PPARg-specific agonist ciglitazone, phytanic acid exerts only minor effects on the differentiation of C3H10T1/2 cells into mature adipocytes. These results clearly demonstrate that phytanic acid acts via different PPAR isoforms to modulate expression of genes involved in glucose metabolism, thus suggesting a potential role of phytanic acid in the management of insulin resistance. PMID- 11923222 TI - DNA oligonucleotide treatment corrects the age-associated decline in DNA repair capacity. AB - Age-related decline in DNA repair capacity (DRC) is associated with decreased constitutive levels of p53 and other nucleotide excision repair proteins. To determine whether pretreatment of cells with small DNA oligonucleotides compensates for decreased DRC in the elderly, fibroblasts from donors of different ages were pretreated with thymidine dinucleotide (pTT), a 5' phosphorylated 9 base oligonucleotide (p9mer) or diluent alone for 48 h, then UV irradiated with solar-simulated light. Western blot analysis revealed age associated decreases of 40%-80% between newborn and old adult donor cells in the constitutive protein levels of p53, p21, XPA, RPA, ERCC1, and PCNA. Treatment with pTT or p9mer up-regulated these proteins by 200%-650% at 24, 48, and 72 h. Moreover, pretreatment with oligonucleotides significantly increased the removal rate of photoproducts as determined by reacting DNA with thymine dimer-specific antibodies: 40+/-5% vs. 20+/-9% and 15+/-11% remained after 24 h in diluent, pTT and p9mer treated cells, respectively. Oligonucleotide-treated adult cells removed thymine dimers at least as rapidly as diluent treated newborn cells, demonstrating that pTT and p9mer completely corrected the age-associated decrease in DRC. Our studies suggest that topical oligonucleotide treatment may enhance DRC in older adults and thus reduce the carcinogenic risk from solar UV irradiation in this age group. PMID- 11923223 TI - Ammonia induces MK-801-sensitive nitration and phosphorylation of protein tyrosine residues in rat astrocytes. AB - Astrocytes play a key role in the pathogenesis of ammonia-induced neurotoxicity and hepatic encephalopathy. As shown here, ammonia induces protein tyrosine nitration in cultured rat astrocytes, which is sensitive to the N-methyl-D aspartate (NMDA) receptor antagonist MK-801. A similar pattern of nitrated proteins is produced by NMDA. Ammonia-induced tyrosine nitration depends on a rise in [Ca2+]i, IkB degradation, and NO synthase (iNOS) induction, which are prevented by MK-801 and the intracellular Ca2+ chelator 1,2-bis(o aminophenoxy)ethane-N,N,N',N'-tetraacetic acid (BAPTA-AM). Moreover, the increase in tyrosine nitration is blunted by L-NMMA, 1400W, uric acid, Cu, Zn-superoxide dismutase/catalase treatment, and methionine-sulfoximine, which indicate the involvement of reactive nitrogen intermediates and intracellular glutamine accumulation. Such reactive nitrogen intermediates additionally mediate ammonia induced phosphorylation of the MAP-kinases Erk-1/Erk-2 and p38MAPK. Among the proteins, which are tyrosine -nitrated by ammonia, glyceraldehyde-3-phosphate dehydrogenase, the peripheral-type benzodiazepine receptor, Erk-1, and glutamine synthetase are identified. Ammonia-induced nitration of glutamine synthetase is associated with a loss of enzymatic activity. Astroglial protein tyrosine nitration is found in brains from rats after acute ammonia-intoxication or after portacaval anastomosis, indicating the in vivo relevance of the present findings. The production of reactive nitrogen intermediates and protein tyrosine nitration may alter astrocyte function and contribute to ammonia neurotoxicity. PMID- 11923224 TI - Effects of altered glucocorticoid sensitivity in the T cell lineage on thymocyte and T cell homeostasis. AB - The homeostatic regulation that controls total thymocyte and peripheral T-cell numbers is not clearly understood. We describe here a direct hormonal influence of endogenous levels of glucocorticoids (GCs) on thymocyte and peripheral T-cell homeostasis independent of indirect systemic effects of GCs. The results were obtained by generating transgenic mice with an altered GC sensitivity targeted to thymocytes and peripheral T cells by increasing or decreasing glucocorticoid receptor (GR) expression specifically in thymocytes and peripheral T cells. A twofold increase in GC sensitivity resulted in a major decrease in thymocyte number, affecting all subpopulations, although single-positive CD8+ cells were less influenced. In the thymus, this was due to increased apoptosis in the organ, whereas proliferation of thymocyte populations was unaffected. In the periphery, a pronounced reduction in T-cell number was seen, demonstrating an effect of endogenous GCs also on T-cell homeostasis. The effects were confirmed in transgenic mice with reduced GR expression, which showed increased thymocyte and T-cell numbers. Thus, our data demonstrate that physiological GC levels are directly involved in controlling the size of both thymocyte and T-cell pools. PMID- 11923225 TI - Preeclampsia is associated with altered Ca2+ regulation and NO production in human fetal venous endothelial cells. AB - Preeclampsia (PE) is a leading cause of maternal hypertension in pregnancy, fetal growth restriction, premature birth, and fetal and maternal mortality (1). Activation and dysfunction of the maternal and fetal endothelium in PE may be the consequence of increased oxidative stress associated with circulating lipid peroxides (2-4), and in cases of severe maternal hypertension, uterine and umbilical artery waveforms are abnormal (5). We have investigated PE-associated abnormalities in the regulation of intracellular Ca2+ ([Ca2+]i) and cyclic guanosine monophosphate (cGMP) production (index of nitric oxide [NO]) in human fetal umbilical vein endothelial cells. Basal [Ca2+]i was slightly elevated in PE cells, whereas agonist-stimulated Ca2+ entry was reduced in cells from PE compared with normal term or age-matched preterm pregnancies. Furthermore, PE cells exhibited a decreased permeability to Ba2+ but an increased permeability to Mn2+ and Gd3+, suggesting that PE is associated with phenotypic alterations in fetal endothelial cation channel(s). Basal and histamine-stimulated cGMP levels were elevated in PE compared with preterm or normal cells, implying an increased NO production in PE. However, immunoblots for endothelial NO synthase (eNOS) and soluble guanylyl cyclase (sGC) revealed reduced eNOS expression in PE and preterm cells, with negligible changes in sGC levels. This study provides important and novel insights into abnormalities of fetal endothelial cells isolated from women with PE, reveal ing an altered cation membrane permeability and activity of eNOS sGC pathway. As these changes are sustained in culture in vitro, this may reflect long-term "programming" of the fetal cardiovascular system. PMID- 11923226 TI - High-resolution scanning patch-clamp: new insights into cell function. AB - Cell specialization is often governed by the spatial distribution of ion channels and receptors on the cell surface. So far, little is known about functional ion channel localization. This is due to a lack of satisfactory methods for investigating ion channels in an intact cell and simultaneously determining the channels' positions accurately. We have developed a novel high-resolution scanning patch-clamp technique that enables the study of ion channels, not only in small cells, such as sperm, but in submicrometer cellular structures, such as epithelial microvilli, fine neuronal dendrites, and, particularly, T-tubule openings of cardiac myocytes. In cardiac myocytes, as in most excitable cells, action potential propagation depends essentially on the properties of ion channels that are functionally and spatially coupled. We found that the L-type calcium and chloride channels are distributed and colocalized in the region of T tubule openings, but not in other regions of the myocyte. In addition, chloride channels were found in narrowly defined regions of Z-grooves. This finding suggests a new synergism between these types of channels that may be relevant for action potential propagation along the T-tubule system and excitation-contraction coupling. PMID- 11923227 TI - Extracellular signal-regulated kinase activation predominantly in Muller cells of retina with endotoxin-induced uveitis. AB - PURPOSE: To evaluate the consequences of mitogen-activated protein kinase (MAPK) mediated signaling in retinas with endotoxin-induced uveitis (EIU). METHODS: EIU was induced with footpad inoculation of lipopolysaccharide (LPS). To identify the expression and activity of extracellular signal-regulated kinase (ERK), c-Jun NH2 terminal kinase (JNK), and p38, Western blot analysis and immunohistochemistry (IHC) were performed using antibodies against these kinases and phosphorylated forms. To evaluate the ERK mRNA expression level, semiquantitative reverse transcription-polymerase chain reaction (RT-PCR) was performed. To identify cell species that express phosphorylated (p)-ERK, simultaneous demonstration of p-ERK and glial fibrillary acidic protein (GFAP) was performed with combined IHC and in situ hybridization. Dexamethasone (Dex) was used to reduce the LPS-induced inflammatory stimulus, and changes in p-ERK expression were evaluated by Western blot analysis after treatment. RESULTS: Only p-ERK among the phosphorylated MAPKs increased after LPS stimulation, according to Western blot analysis. p-ERK increased after LPS injection, whereas both the Western blot and RT-PCR studies showed no apparent changes in ERK-1 and -2 expression. IHC revealed that strong p ERK-positive staining initially appeared in the Muller cell bodies. Thereafter p ERK immunostaining was also observed transiently in the radial processes of the Muller cells. The double-labeling study revealed that almost all Muller cells were positive for GFAP and p-ERK. Dex treatment substantially reduced expression of p-ERK, beginning 12 hours after treatment. CONCLUSIONS: The present study suggests that LPS stimulation activates ERK in Muller cells, whereas the total amount of ERK is unchanged. Because the LPS-induced p-ERK level was reduced by Dex treatment, its expression seems to be associated with ocular inflammatory stimulus. Because the inflammatory stimulus elicited in EIU upregulated ERK activity in Muller cells, activated Muller cells may play a crucial role in protecting retinal cells from such inflammation. PMID- 11923228 TI - Presence of a fluid-conducting meshwork in xenografted cutaneous and primary human uveal melanoma. AB - PURPOSE: Recently, it was reported that tumor cells themselves generate channels and networks in three-dimensional culture and can be found lining channels (some containing red blood cells [RBCs]) in vivo, and they express endothelial or vascular genes in aggressive uveal melanoma. The implications of these data for current insights in the involvement of angiogenesis in tumor growth, metastasis and therapeutic intervention are considerable. Therefore, this possibility was investigated in the current study. METHODS: Thirty human uveal melanomas and 20 xenografts of human cutaneous melanoma were analyzed by Azan histochemistry and immunostaining of endothelial markers. Additionally, in xenografted tumors a tracer study was performed with confocal microscopy and immunoelectron microscopy. RESULTS: Lumina or spaces without endothelial lining containing RBCs were not detected in any lesion. Functional evaluation of the vasculature in xenografts demonstrated rapid tracer appearance both inside and outside blood vessels. Outside blood vessels it spread along matrix networks of arcs and back to-back loops. Confocal microscopy showed that this extracellular matrix was deposited as stromal sheets around nests of tumor cells. Laminin immunostaining revealed that between sheets surrounding adjacent nests, spaces were present. These spaces were filled, however, with collagen and different types of cells, including cells stained for macrophage markers. CONCLUSIONS: Although no evident endothelium-free and RBC-containing channels were present in the tissues examined, there are fluid-conducting spaces in the form of stromal sheets between nests of tumor cells. In this stromal network, blood vessels are embedded. The authors postulate that this extracellular matrix tissue represents a fluid conducting meshwork. PMID- 11923229 TI - Preservation of retinal morphology and functions in royal college surgeons rat by nilvadipine, a Ca(2+) antagonist. AB - PURPOSE: The Royal College of Surgeons (RCS) rat is the most extensively studied animal model for understanding the molecular pathology in inherited retinal degeneration, such as retinitis pigmentosa (RP). The purpose of the present study was to evaluate the pharmacologic effects of several Ca(2+) antagonists on the retinal degeneration of RCS rats. METHODS: Several Ca(2+) antagonists, diltiazem, nicardipine, nilvadipine, and nifedipine, were intraperitoneally administered and retinal morphology and functions analyzed. RESULTS: Among the Ca(2+) antagonists, only intraperitoneally administered nilvadipine preserved retinal morphology and electroretinogram responses in RCS rats during the initial stage of retinal degeneration. Studies using immunohistochemistry, RT-PCR, and Western blot analysis revealed significant enhancement of rhodopsin kinase and alphaA crystallin expression and suppression of caspase 1 and 2 expression in the retina of nilvadipine-treated rats. CONCLUSIONS: These data suggest that nilvadipine is beneficial for the preservation of photoreceptor cells in RCS rats and can be used to treat some patients with RP. PMID- 11923230 TI - Assessment of the impact of vision impairment. AB - PURPOSE: To describe the psychometric characteristics of the Impact of Vision Impairment (IVI) Profile and evaluate its validity and reliability over time and between different forms of administration. METHODS: The IVI is a 32-item questionnaire developed to measure the impact of vision impairment on restriction of participation in daily activities in five domains of functioning. Each item is rated on a six-level scale from "no difficulty" to "can't do because of vision." The IVI was administered by trained interviewers to 115 people with impaired vision (visual acuity less than 6/12 or visual field deficit) who attended the Royal Victorian Eye and Ear Hospital, a vision rehabilitation agency, or a self help group for people with impaired vision. Data were also collected on demographic characteristics of participants, cause of vision impairment, and distance and near vision. General health status was assessed with the Short Form 12 (SF-12) of the Physical and Mental Health Summary Scales. A subset of participants completed the IVI twice, either 1 to 2 weeks apart or by different forms of administration (different interviewers or self). RESULTS: Internal consistency of total and domain average IVI scores was high (alpha = 0.80-0.96) and sequential elimination of items did not affect consistency. Total and domain average IVI scores correlated moderately with both near and distance vision (r = 0.21-0.31) but did not correlate with physical or general health or comorbidity. Total and domain average IVI scores correlated most closely with global measures of restriction of participation (r = 0.44-0.82). Principal-components analysis confirmed that all IVI items contribute to one underlying theme and tended to confirm two of the five domains: emotional reaction to vision loss and mobility. The first three components explained 43%, 8%, and 6% of the variation in the data. Guttman split-half reliability coefficients between different forms of administration and over time ranged from 0.73 to 0.94 for domain and total IVI scores. Mean absolute difference for domain and total scores between administrations was less than 1 step for all domains and the total score. CONCLUSIONS: This study provides support that the IVI has sufficient internal and construct validity to measure the effect of vision impairment on restriction of participation in daily activities. The IVI demonstrates acceptable reliability over a short period and yields consistent results between interviewers. The IVI can also be self-administered with assurance that the results will be comparable to those that would have been obtained by a trained interviewer. Therefore, the psychometric characteristics of the IVI support its use in assessment of the vision rehabilitation needs of people with impaired vision. Its stability over time indicates that it has potential to evaluate outcomes of intervention. PMID- 11923231 TI - Longitudinal study of trachomatous trichiasis in The Gambia: barriers to acceptance of surgery. AB - PURPOSE: Investigation of compliance with surgery for trachomatous trichiasis has become a priority of the World Health Organization. This study was conducted to investigate attitudes toward trichiasis and its treatment and to determine the rate of surgical uptake in The Gambia. METHODS: A 1-year longitudinal study was performed in 190 subjects with trichiasis. Persons with major trichiasis (involving five lashes or more) were referred for surgery, and those with minor trichiasis were advised to epilate. Outcome measures included attitudes toward trichiasis and its treatment, reported barriers to surgical uptake, acceptance rates for surgery, and factors affecting acceptance. RESULTS: Twenty-three percent (95% confidence interval [CI] 16.5%-30.6%) of subjects with major trichiasis attended for surgery during the year. Degree of ignorance about surgery, symptoms impeding work, and a multiple income source for the head of household predicted attendance. Reported lack of time predicted nonattendance. Sixty-eight percent of patients who had undergone surgery were trichiasis free at last follow-up. CONCLUSIONS: Poor attendance for surgery remains a problem in The Gambia. Barriers include ignorance and lack of time and money. Health education and surgical delivery strategies are needed to overcome these barriers. Regular audit of surgical results is necessary, with retraining where indicated. PMID- 11923232 TI - Analysis of customized corneal ablations: theoretical limitations of increasing negative asphericity. AB - PURPOSE: To determine the ablation depths of customized myopic excimer laser photoablations performed to change corneal asphericity after laser in situ keratomileusis (LASIK) and photorefractive keratectomy (PRK). METHODS: A mathematical model of aspheric myopic corneal laser surgery was generated. The initial corneal surface was modeled as a conic section of apical radius R(1) and asphericity Q(1). The final corneal surface was modeled as a conic section of apical R(2) and asphericity Q(2), where R(2) was calculated from the paraxial optical formula for a given treatment magnitude (D), and Q(2) was the intended final asphericity. The aspheric profile of ablation was defined as the difference between the initial and final corneal profiles for a given optical zone diameter (S), and the maximal depth of ablation was calculated from these equations. Using the Taylor series expansion, an equation was derived that allowed the approximation of the central depth of ablation (t(0)) for various magnitudes of treatment, optical zone diameters, and asphericity. In addition to the Munnerlyn term (M), incorporating Munnerlyn's approximation (-D small middle dot S(2)/3), the equation included an asphericity term (A) and a change of asphericity term (Delta). This formula (t(0) = M + A + Delta) was used to predict the maximal depth of ablation and the limits of customized asphericity treatments in several theoretical situations. RESULTS: When the initial and final asphericities were identical (no intended change in asphericity; Q(1) = Q(2); Delta = 0), the maximal depth of ablation (t(0) = M + A) increased linearly with the asphericity Q(1). To achieve a more prolate final asphericity (Q(2) < Q(1); dQ < 0; Delta > 0), the maximal depth of ablation (M + A + Delta) was increased. For treatments in which Q(2) was intended to be more oblate than Q(1) (Q(2) > Q(1); dQ > 0; Delta < 0), the maximal depth of ablation was reduced. These effects sharply increased with increasing diameters of the optical zone(s). Similarly, in the case of PRK, the differential increase in epithelial thickness in the center of the cornea compared with the periphery resulted in increased oblateness. CONCLUSIONS: Aspheric profiles of ablation result in varying central depths of ablation. Oblateness of the initial corneal surface, intentional increase in negative asphericity, and enlargement of the optical zone diameter result in deeper central ablations. This may be of clinical importance in planning aspheric profiles of ablation in LASIK procedures to correct spherical aberration without compromising the mechanical integrity of the cornea. PMID- 11923233 TI - BIGH3 mutation spectrum in corneal dystrophies. AB - PURPOSE: To investigate the molecular pathology underlying BIGH3-related corneal dystrophies (CDs) and to further delineate genotype-phenotype specificity. METHODS: Sixty-one index patients with CDs were subjected to phenotypic and genotypic characterization. The corneal phenotypes of all patients were assessed by biomicroscopy and documented by slit lamp photography. The BIGH3 gene was amplified exon by exon from constitutional DNA to perform single-strand conformation polymorphism (SSCP) analysis, followed by direct bidirectional sequencing of abnormal conformers. RESULTS: The phenotypes of CDs were classified as lattice CD in 30 patients, Groenouw type I in 12 (CDGGI), Avellino in 7 (CDA), Reis-Buckler in 8 (CDRB), and Thiel-Behnke in 4 (CDTB). Fifty occurrences of 16 distinct mutations were identified, including 8 novel mutations responsible for lattice type IIIA in three patients (CDLIIA), intermediate type I/IIIA (CDLI/IIIA) in four patients, and atypical CDL with deep deposits in one patient (CDL-deep). CONCLUSIONS: Disease-causing mutations were identified in 80% of the patients (50/61). All mutations localize in two regions of kerato-epithelin: the amino acid R124 and BIGH3 fasc domain 4. This study also confirms the mutation hot spot at positions R124 and R555 with nearly 50% of the mutations targeting these two amino acids (24/50). In addition the corneal phenotypes induced by changes at R124 and R555 are amino acid specific: R124C in CDLI, R555W and R124S in CDGGI, R124H in CDA, R124L in CRRB, and R555Q in CDTB. In CDLIIIA, CDLI/IIIA, and CDL-deep the genotype-phenotype correlation is domain specific, with all changes occurring at the boundary or within the fasc4 domain. PMID- 11923234 TI - Expression of integrins and MMPs during alkaline-burn-induced corneal angiogenesis. AB - PURPOSE: To determine in a corneal alkaline burn model of angiogenesis whether the expression of integrins and MMPs is consistent with a VEGF-induced angiogenic response. METHODS: Neovascularization in female Sprague-Dawley rats was induced by alkaline cauterization of the central cornea. RT-PCR for integrins alpha(1), alpha(2), beta(3), and beta(5); the endothelial marker CD31; and metalloproteinases MMP-2 and MT1-MMP was performed on naive corneas and on cauterized corneas 72 and 288 hours after cautery. Analyses of protein and MMP expression were conducted on naive corneas and on cauterized corneas 24, 72, 120, and 168 hours after cautery by immunofluorescence microscopy and gelatin zymography. RESULTS: RT-PCR indicated a correlation between the induced angiogenic response and the expression of alpha(1) and beta(3) integrin subunits and MT1-MMP. Immunohistochemical analysis indicated that alpha(1), alpha(2), alpha(5), and beta(5) integrins and MMP-2 and MT1-MMP were expressed on the newly developing vasculature. The beta(3) integrin was preferentially expressed on platelets. CONCLUSIONS: Integrin expression during neovascularization of rat corneas in response to alkaline injury correlates with an angiogenic response that uses the VEGF/alpha(v)beta(5) pathway. MMP-2 and MT1-MMP, but not MMP-9, are expressed in a pattern consistent with their involvement in the angiogenic response. PMID- 11923235 TI - Identification of insulin in the tear film and insulin receptor and IGF-1 receptor on the human ocular surface. AB - PURPOSE: Insulin produces pleiotropic effects on sensitive tissues, including the ocular surface, through the tyrosine kinase insulin receptor. Cerebrospinal fluid and secreted fluids, such as milk and saliva, have been reported to contain insulin. In the present study, the presence of insulin was examined in tear film, and the expression of insulin and insulin-like growth factor (IGF)-1 receptor was examined in the human cornea and conjunctiva. METHODS: Stimulated tear samples collected from 33 volunteers (17 men, 16 women), aged 23 to 51 years, who were fed or fasted for 12 hours, were assayed for total protein and insulin content by the biuret dye test and a radioimmunoassay, respectively. Frozen sections of human cornea (n = 4) and conjunctiva (n = 3) were incubated with anti-insulin receptor and anti-IGF-1 receptor antibodies and developed with a secondary antibody-peroxidase conjugate. RESULTS: Insulin was detected in all tear samples analyzed, the mean concentration being 0.404 +/- 0.129 ng/mL. There were no gender-related differences. In fed subjects, tears tended toward a higher insulin content than those in fasted individuals. There was no linear correlation between insulin and total protein content (mean, 4.61 +/- 0.79 mg/mL) in the tear film. Insulin and IGF-1 receptors were detected in the plasma membrane and cytoplasm of corneal and conjunctival epithelial cells. CONCLUSIONS: To the best of the authors' knowledge, this study represents the first demonstration of insulin in human tear film and the presence of insulin and IGF-1 receptor on the human ocular surface. These results suggest that the pancreatic hormone may play a metabolic and/or mitogenic role on the ocular surface. PMID- 11923236 TI - Retroviral gene therapy vectors for prevention of excimer laser-induced corneal haze. AB - PURPOSE: To determine the in vivo efficacy and safety of a retroviral vector bearing an antiproliferative dominant negative mutant cyclin G1 (dnG1) construct, when used for the prevention of corneal haze after phototherapeutic keratectomy (PTK). METHODS: For in vivo efficacy studies, a 6-mm-diameter, 150-microm-deep transepithelial PTK, performed with a clinical 193-nm ArF excimer laser (VISX Star2, Santa Clara, CA) was performed on the left eyes of 20 adult New Zealand White rabbits. The surgically altered eyes were subsequently treated with eye drops containing: a retroviral vector bearing a dnG1 construct (dnG1; n = 7), a control retroviral vector (null vector) bearing only the neomycin resistance, neo(r), gene (n = 7), or a retroviral vector bearing an antisense cyclin G1 (aG1) construct (n = 6). The time of closure of the corneal epithelial defect was monitored daily with fluorescein staining. Corneal haze was evaluated before surgery and at 2, 3, and 4 weeks after surgery, with a digital imaging system. Biodistribution studies for detection of potential vector dissemination to nontarget organs were conducted by PCR-based assay. RESULTS: The re epithelialization rate was similar among treatment groups, with complete closure of the corneal epithelial defect at 72 hours (P > 0.05). Significant corneal haze developed in the null and aG1 vector-treated groups (P 10 years' duration) and 20 corneas of 20 age-matched (+/-5 years) control subjects who had never worn contact lenses, were examined by confocal microscopy in vivo. The contact lens wearers removed their lenses 12 to 24 hours before the examination. Full-thickness images were recorded from the central and temporal cornea, and bright objects (keratocyte nuclei) in images were manually counted to calculate keratocyte density. Stromal intensity (backscatter) was measured by calculating the mean grayscale value (corrected for camera and light source variations) from the center of stromal images. Epithelial thickness was determined from the distance between images of the surface epithelium and subbasal nerve plexus. Central corneal sensitivity was measured by Cochet-Bonnet esthesiometry and correlated with the number of nerve fiber bundles in the subbasal nerve plexus. RESULTS: Full-thickness central and temporal keratocyte densities in contact lens wearers were 22,122 +/- 2,676 cells/mm(3) (mean +/- SD) and 20,731 +/- 2,627 cells/mm(3), respectively, and were not significantly different from central and temporal keratocyte densities in control subjects (P = 0.29). The minimum detectable difference in cell density was 11% (2346 cells/mm(3) and 2235 cells/mm(3) in central and temporal stroma, respectively). Temporal epithelial thickness was 46.3 +/- 4.7 microm in contact lens wearers and 50.9 +/- 4.7 microm in control subjects (P = 0.02). Central epithelial thickness and stromal backscatter did not differ between contact lens wearers and control subjects (P > 0.05). Corneal sensitivity was lower in contact lens wearers than it was in control subjects (P = 0.05) and did not correlate with the number of nerve fiber bundles in the subbasal nerve plexus. CONCLUSIONS: Long-term daily contact lens wear and its associated stromal hypoxia and acidosis have no demonstrable effect on keratocyte density. The temporal epithelium is thinner in corneas of long-term contact lens wearers than in control subjects. Decreased corneal sensitivity in contact lens wearers is not accompanied by decreased nerve fiber bundle density. PMID- 11923240 TI - Decreased levels of the goblet cell mucin MUC5AC in tears of patients with Sjogren syndrome. AB - PURPOSE: To determine whether the relative amounts of mucin mRNA in the conjunctival epithelium and mucin protein in the tears are altered in patients with Sjogren syndrome compared with healthy individuals. METHODS: Tear fluid was collected from the inferior fornix of normal subjects (n = 17) and patients with Sjogren syndrome (n = 11) after instillation of 60 microL sterile water onto the ocular surface. Immediately after tear fluid collection, conjunctival epithelium was obtained by filter paper-stripping from the bulbar temporal region for mRNA isolation. Primers to nontandem repeat sequences of the gel-forming mucin MUC5AC and the membrane-spanning mucins MUC1 and MUC4 were used in real-time RT-PCR to determine relative abundance of MUC mRNA in patients with Sjogren syndrome in relation to that of normal subjects. Enzyme-linked immunosorbent assay was performed on neuraminidase-treated tears, using a polyclonal antibody against a synthetic peptide mimicking the deduced amino acid sequence from the D3 region of MUC5AC. RESULTS: The number of RNA transcripts for the goblet cell-specific mucin MUC5AC in the conjunctival epithelium of patients with Sjogren syndrome was significantly lower than in normal individuals. No significant changes were detected when analyzing the mRNA levels of the mucins expressed by the stratified epithelium of the conjunctiva, MUC1 and MUC4. Protein levels of the goblet cell mucin MUC5AC were significantly reduced in the tear fluid of patients with Sjogren syndrome, corroborating mRNA data obtained using real-time RT-PCR. CONCLUSIONS: The tear fluid of patients with Sjogren syndrome has reduced levels of the goblet cell-specific mucin MUC5AC, which correlates to decreased levels of conjunctival MUC5AC mRNA. The authors propose that deficiency of MUC5AC mucin in tears constitutes one of the mechanisms responsible for tear film instability in Sjogren syndrome. PMID- 11923242 TI - Impact of rigid gas-permeable contact lens extended wear on corneal epithelial barrier function. AB - PURPOSE: To measure the effect of hypoxia and eye closure on epithelial permeability to fluorescein (P(dc)) during rigid lens extended wear (EW). METHODS: Central corneal thickness (CT) and P(dc) were measured in 42 subjects with an optical pachometer and automated scanning fluorophotometer, respectively. All subjects had been successfully wearing rigid gas-permeable (RGP) lenses on a 6-night EW regimen, and each individual was randomized to wear either medium- or high-oxygen-permeable (Dk) RGP lenses (two types of siloxane-fluorocarbon polymer lenses with Dk of 49 and 92). CT and P(dc) measurements were performed at an afternoon visit (baseline) and were repeated in the morning after 8 hours of overnight wear. Subjects slept with a patch over the right eye. The patch was not removed until immediately before the morning measurement. RESULTS: The mean overnight swelling response for subjects in the medium-Dk group was greater than that in the high-Dk group. Results of a paired t-test indicate that the eye wearing the medium-Dk lens with a patch overnight had a significant increase in epithelial permeability. Results of mixed-effect models suggest that eye closure and lens-induced hypoxia are significant factors in altering P(dc). CONCLUSIONS: The results indicate that corneal epithelial permeability increases with hypoxic dose and that epithelial barrier function is impaired by overnight rigid lens wear. PMID- 11923241 TI - Effect of alloantibodies on corneal allograft survival. AB - PURPOSE: The precise role of antibodies in corneal transplantation is ambiguous, with evidence to support as well as repudiate their involvement in graft rejection. Accordingly, this study was undertaken to investigate the direct contribution of donor-specific antibodies to corneal graft rejection. METHODS: Serum samples from CB6F1 rejecters of orthotopically grafted C3H/Hej corneas were tested by ELISA for elevated levels of donor-specific alloantibody. Orthotopic corneal allograft rejection was also examined in B-cell-deficient mice. In a prospective study, naive BALB/c T-cell-deficient nude mice and complement depleted nude mice were passively infused with immune donor-specific serum and grafted with fully allogeneic C57BL/6J corneas. The incidence and speed of graft rejection were observed in each case. The susceptibility of corneal cells to antibody-mediated lysis was tested in vitro. RESULTS: Seventy percent of the CB6F1 hosts that rejected the C3H/Hej corneal allografts possessed significantly elevated levels of alloantibody in serum. Although BALB/c corneal allografts were rejected by B-cell-deficient mice at the same incidence as wild-type control mice, their mean survival time (MST) was significantly longer than that of their wild-type counterparts. Serum of BALB/c mice immunized against C57BL/6J alloantigens produced complement-dependent cytolytic activity against C57BL/6J corneal cells in vitro. Passive transfer of this alloantiserum to T-cell deficient BALB/c nude mice produced complement-dependent corneal lesions, resulting in significantly increased opacity of C57BL/6J corneal grafts, compared with the relatively clear grafts in control hosts. CONCLUSIONS: Alloantibody, although not necessary for corneal graft rejection, can produce extensive injury to corneal allografts in a complement-dependent manner. PMID- 11923243 TI - Keratocyte activation and apoptosis in transplanted human corneas in a xenograft model. AB - PURPOSE: To study keratocyte activation and cellular apoptosis in transplanted human corneas during the early postoperative period. METHODS: Ten human donor corneas preserved for 6 days at 4 degrees C were transplanted into the eyes of 10 adult cats. After confocal and specular microscopy in vivo 1 week after keratoplasty, the cats were killed, and the fixed corneas were examined by TUNEL assay and by scanning (SEM) and transmission electron microscopy (TEM). RESULTS: Abnormal keratocytes, in which portions of cell bodies and processes as well as nuclei were visible, were present in all corneas and occupied the anterior 16 to 562 microm of the stroma. By TEM in the same corneas, these abnormalities represented keratocytes that were activated to a repair phenotype. Only 0% to 1% of all corneal cells were apoptotic by TUNEL assay, except for the donor keratocytes near the wound, where 7% were apoptotic. The midstromal keratocyte density was decreased at 13,936 +/- 5,910 cells/mm(3) (mean +/- SD), and the endothelial cell density was 2,298 +/- 688 cells/mm(2), representing an endothelial cell loss of 7% +/- 16%. CONCLUSIONS: Substantial keratocyte activation and low levels of cellular apoptosis occur 1 week after human corneal transplantation. The human-to-cat xenograft model of corneal transplantation demonstrated endothelial cell loss and other clinical findings similar to human allografts. The model will be useful for preclinical testing of new methods of long-term corneal preservation and of donor endothelial cell augmentation, as well as the study of human corneal wound healing and keratocyte replacement during the early postoperative period. PMID- 11923244 TI - A novel method to evaluate residence time in humans using a nonpenetrating fluorescent tracer. AB - PURPOSE: The purpose of these investigations was to develop an improved method for measuring precorneal residence time (RT) and to demonstrate its efficacy with novel formulations. METHODS: A biomicroscope was adapted for use as a clinical fluorometer. Using a nonpenetrating fluorescent probe (FITC-dextran, 70,000 73,000 molecular weight [MW]), RT was estimated as the time to return to baseline (gross RT) and from parameters derived from least-squares regression fits to the decay data (area under the curve [AUC], elimination rate, and time for 50% of the signal to be eliminated [T(50)]). One rabbit and two human studies were conducted. The studies were randomized, double-masked, and controlled. Repeatability in humans was examined in 15 subjects (six determinations per subject, n = 90 total). RESULTS: The FITC-dextran tracer did not penetrate into corneal tissue. The rabbit gross RTs were 14.5, 15.0, and 16.0 minutes for three low-viscosity solutions (eta = 2.7-7.7 mPa/sec) and 22.5 minutes for a more viscous solution (eta = 357 mPa/sec). For a high-viscosity (eta approximately equal 30,000 mPa/sec) gel in humans, the method demonstrated approximately a twofold increase in gross RT and AUC compared with buffered saline. Repeatability of the method appeared acceptable, with intersubject variability the most significant factor affecting precision. CONCLUSIONS: The new method is safe and convenient and offers comprehensive RT data. Furthermore, it appears to differentiate among formulations. However, as with other tear-influenced parameters, there is significant variability. Thus, sufficient sample sizes are necessary for meaningful comparative investigations. PMID- 11923245 TI - Binocular coordination of saccades in children with strabismus before and after surgery. AB - PURPOSE: To examine the quality of binocular coordination of saccades in children with various types of strabismus and the effect of strabismus surgery. METHODS: Eight subjects were tested (5-15 years old): five with convergent strabismus, three with divergent strabismus. A standard saccade paradigm was used to elicit horizontal saccades to target LEDs (5 degrees to 15 degrees ). Saccades from both eyes were recorded simultaneously with the photograph-electric Skalar IRIS device (Delft, The Netherlands). This task was run before and about 3 weeks after strabismus surgery. RESULTS: Before surgery, the difference in the amplitude of the saccade between the left eye and the right eye was larger (15% of the saccade size) than in normal children of similar age. After strabismus surgery for all subjects the squint angle was reduced, and the amplitude of the disconjugacy of saccades decreased significantly, dropping to normal values (6%). As in normal children, postsaccadic eye drift (both its conjugate and its disconjugate components) was small in amplitude. The difference compared with normal subjects was that disconjugate drift did not restore the disconjugacy of the saccade itself (e.g., in normal subjects drift is convergent when saccade disconjugacy is divergent and vice versa). Rather, disconjugate drift tended to drive the eyes toward static eye misalignment (e.g., the drift was mostly convergent for convergent strabismics and divergent for divergent strabismics). Surgery had no significant effect on either component of the drift. CONCLUSIONS: The improvement of the binocular coordination of the saccades could be due, at least partially, to central adaptive mechanisms rendered possible by surgical realignment of the eyes. Separate mechanisms control the binocular coordination of saccades and the alignment of the eyes during the postsaccadic fixation period. PMID- 11923246 TI - Transcriptional profile of rat extraocular muscle by serial analysis of gene expression. AB - PURPOSE: Although extraocular muscle (EOM) is skeletal muscle, aspects of its biology are unlike other striated muscle. In this study, the broad molecular genetics profile underlying the novel EOM phenotype was examined. METHODS: Serial analysis of gene expression (SAGE) was used to quantify adult rat EOM gene transcripts. SAGE isolates and sequences 10-bp tags from defined locations in mRNA-derived cDNA. Tag sequence-location was used to extract transcript identity from a curated SAGE database, and detection frequencies reflected abundance of corresponding mRNAs. RESULTS: The 54,764 expressed sequence tags generated and sequenced from EOM included 17,602 unique tags. Of the unique tags, 7.8% were detected at high to intermediate levels (>or=5 copies), 19.3% at lower levels (2 4 copies), and 72.9% as single copies; 40% of the tags matched known expressed sequence tags (ESTs), most of which (85.7%) represented a unique EST. Tags without matches in the SAGE database and those expressed as single copies only were not considered further. SAGE tags expressed at more than 0.1% of total transcripts reflected several aspects of muscle biology, including sarcomeric structure, energy metabolism, and ribosomal protein expression. Genes highly expressed in EOM were compared with other existing muscle expression databases to identify conserved and novel patterns in EOM. CONCLUSIONS: The data provide a normative gene expression database and a novel molecular signature that will facilitate study of EOM development and function and of the mechanisms behind its preferential targeting or sparing in neuromuscular disease. PMID- 11923247 TI - Correlation of the binocular visual field with patient assessment of vision. AB - PURPOSE: To determine which measures of the binocular visual field correlate best with the patient's assessment of vision. METHODS: Esterman binocular visual field testing and four other binocular visual field tests (designated peripheral 20 dB [p20], peripheral 22 dB [p22], central 24 dB [c24] and central 26 dB [c26]) were performed in 101 patients with glaucoma or suspected glaucoma. Scores from these five tests, as well as binocular visual field scores calculated from monocular testing (best-location summation and probability summation), were correlated with performance on the National Eye Institute's Visual Function Questionnaire (VFQ) 25 and Short-Form (SF)-36 quality of life instruments, as well as with the linear rating scale utility test. RESULTS: The mean percentage of correct responses was 87%, 69%, 59%, 78%, and 71% for the Esterman, p20, p22, c24, and c26 tests, respectively. The distribution of scores was much broader for the p20 and p22 tests than for the Esterman test. The mean decibels for the binocular visual fields calculated from the monocular visual fields were 21.5 +/- 7.7 dB for the best-location algorithm and 25.1 +/- 6.7 dB for the probability-summation algorithm. The binocular visual field score calculated with the best-location algorithm correlated better with the overall, general vision, distance activities, and peripheral vision domains of theVFQ-25 (partial correlation coefficients of 0.48, 0.48, 0.49, and 0.51, respectively) than did the probability-summation algorithm and all five binocular visual field tests. The best-location algorithm also had the strongest correlation with the linear rating scale utility test (partial correlation coefficient, 0.40). CONCLUSIONS: In this sample of clinic-based patients with glaucoma or suspected glaucoma, a global score derived from a combination of two monocular fields correlated better with patient assessment of vision than did the Esterman and four novel binocular visual field tests. PMID- 11923248 TI - Distribution of myocilin and extracellular matrix components in the juxtacanalicular tissue of human eyes. AB - PURPOSE: To examine ultrastructurally the composition of extracellular matrix (ECM) materials, the distribution of myocilin, and the colocalization of myocilin with ECM components in the juxtacanalicular tissue (JCT) of normal human eyes. METHODS: Postembedding immunoelectron microscopic studies were performed with antibodies specific for major ECM components, including fibronectin, laminin, vitronectin, tenascin, elastin, fibrillin-1, microfibril-associated glycoprotein (MAGP)-1, decorin, versican, and five types of collagen (I, III, IV, V, and VI). Hyaluronic acid was localized with the use of biotinylated hyaluronic acid binding protein. Colloidal gold labeling was also performed using an anti-human myocilin polyclonal antibody. Colocalization of myocilin with ECM components was examined by double labeling, using different-sized gold particles. The possible interaction between myocilin and ECM molecules was evaluated by in vitro binding assays. RESULTS: Amorphous basement membrane-like materials in the JCT were confirmed to be made up chiefly of collagen type IV, laminin, and fibronectin. Elastin was localized to the central core of sheath-derived plaques. Fibronectin, fibrillin-1, MAGP-1, decorin, and type VI collagen were all localized to clusters of the banded material in the sheath surrounding the core, where several types of collagen, glycoproteins, and proteoglycans were also detected. Myocilin was found to associate mainly with the sheath material, overlapping extensively in distribution with fibronectin, fibrillin-1, and MAGP-1 and moderately with decorin and type VI collagen. Its localization was distinct from that of elastin. Interactions of myocilin with molecules such as fibronectin and fibrillin-1 were confirmed biochemically. CONCLUSIONS: This study illustrated ultrastructurally the composition of ECM materials in the JCT of normal human eyes. The key finding was the association of myocilin with microfibrillar architecture in sheath derived plaques where pathologic changes have been documented to occur in eyes of patients with primary open-angle glaucoma. PMID- 11923250 TI - Immunohistologic evidence for retinal glial cell changes in human glaucoma. AB - PURPOSE: Glial cells are structurally and functionally linked to neuronal tissues. Pathologically, the cells may be activated and characterized by increased size and number and altered cellular properties. In glaucoma, pathologic mechanisms within the anterior optic nerve may include glial activation. This study examines morphologic changes of glial cells in the retinas of glaucomatous eyes compared with age-matched control retinas. METHODS: Paraffin processed or flatmounted retinas from 17 human donor eyes [7 normal (donor age, 87.3 +/- 8.3 years) and 10 glaucomatous (donor age, 87.1 +/- 6.9 years)] were examined. With immunohistochemical methods, retinal glial cells were stained with an antibody to glial fibrillary acidic protein (GFAP). The morphology of the glial cells in normal and glaucomatous retinas was evaluated with fluorescence microscopy. RESULTS: Three types of glial cells were identified in flatmounted retinas with differing distributions in the peripapillary region, the nerve fiber layer (NFL), and along the capillaries. Compared with normal eyes, in glaucomatous retinas the glial cells in the peripapillary region showed an increase in density and exhibited a deformation of the end feet. The astrocytes distributed among the NFL showed little difference from normal. The astrocytes accompanying the capillary bed showed a redistribution in the glaucomatous retinas. The quantification of glial cells in paraffin-processed glaucomatous retinas exhibited a profound increase in density and a significant increase of GFAP immunoreactivity in contrast to the lightly stained glial cells in normal retinas. CONCLUSIONS: The activation of glial cells in the glaucomatous retina was characterized in changes of intensity of GFAP immunoreactivity and morphology around the larger blood vessels, compared with age-matched normal retinas. The relationships between glial cells, neuronal cells, and the vasculature, as well as the potential role of glial cells in pathologic mechanisms during different stages of neuronal damage in glaucoma, are discussed. PMID- 11923249 TI - Caspase activation and amyloid precursor protein cleavage in rat ocular hypertension. AB - PURPOSE: Retinal ganglion cell (RGC) death in glaucoma involves apoptosis. Activation of caspases and abnormal processing of amyloid precursor protein (APP) are important events in other chronic neurodegenerations, such as Alzheimer's disease (AD). The retinal expression and activation of caspases and the patterns of caspase-3-mediated APP processing in ocular hypertensive models of rat glaucoma were investigated. METHODS: RGC death was produced in one eye by chronic exposure to increased intraocular pressure (IOP) or by optic nerve transection. Elevated IOP was produced by obstruction of aqueous humor outflow with laser coagulation or limbal hypertonic saline injection. Caspase activity and APP processing in the retina were examined by RNase protection assay (RPA), immunocytochemistry, immunoblot assay, and colorimetric assay. RESULTS: RPA revealed elevations of caspase-3 mRNA, as well as other apoptosis-related mRNAs. Immunocytochemistry showed caspase-3 activation in RGCs damaged by ocular hypertension. The generation of the caspase-3-mediated APP cleavage product (DeltaC-APP) was also increased in ocular hypertensive RGCs. Western immunoblot assay and colorimetry revealed significantly more activated caspase-3 in ocular hypertensive retinas than in control retinas. The activated form of caspase-8, an initiator caspase, and amyloid-beta, a product of APP proteolysis and a component of senile plaques in AD, were detected in RGCs by immunohistochemistry significantly more often in ocular hypertensive than in control retinas. The amounts of full-length APP were reduced and amyloid-beta-containing fragments were increased in ocular hypertensive retinas by Western immunoblot assay. CONCLUSIONS: Rat RGCs subjected to chronic ocular hypertension demonstrate caspase activation and abnormal processing of APP, which may contribute to the pathophysiology of glaucoma. PMID- 11923251 TI - Effect of administration of CTLA4-Ig as protein or cDNA on corneal allograft survival. AB - PURPOSE: To examine the role of the CD28-CD80-CD86 pathway of T-lymphocyte costimulation in corneal allograft rejection and the effect of blockade of that pathway on graft survival. METHODS: Kinetics of CD80 and CD86 expression in the cornea and draining lymph nodes were examined by RT-PCR and immunohistochemistry in untreated allograft recipients in a high-responder rat model. The effect of blockade of CD28-mediated costimulation was first examined by ex vivo incubation of excised Brown Norway rat donor cornea with the inhibitory protein CTLA4-Ig or an adenovirus vector (AdCTLA) expressing CTLA4-Ig, before grafting into Lewis rat recipients. A second group of graft recipients received systemic posttransplantation treatment with either CTLA4-Ig or AdCTLA. RESULTS: Expression of CD80 mRNA was increased in both donor and recipient cornea 16 hours after transplantation, whereas CD86 was detected constitutively, with no significant early increase. Immunohistochemistry on day 5 after transplantation demonstrated major histocompatibility complex (MHC) class II expression, no CD80, and only a trace of CD86 in corneal allografts. In lymph nodes strong MHC class II, weak CD80, and moderate CD86 expression was noted. Both donor cornea and recipient treatment with CTLA4-Ig resulted in prolonged allograft survival. AdCTLA was found to induce sustained secretion of bioactive CTLA4-Ig from corneas infected ex vivo. Survival of corneal allografts incubated with AdCTLA was marginally prolonged, and systemic treatment with AdCTLA significantly prolonged survival. CONCLUSIONS: Protein- or gene-based administration of CTLA4-Ig prolongs allograft survival by treatment of either the recipient or the donor tissue ex vivo before grafting. PMID- 11923252 TI - Extensive deposits of complement C3d and C5b-9 in the choriocapillaris of eyes of patients with diabetic retinopathy. AB - PURPOSE: To determine the presence of activated complement components in eyes affected by diabetic retinopathy. METHODS: Eyes of 50 deceased donors with diabetic retinopathy and of 10 nondiabetic subjects with uveal melanoma (n = 6) or phthisical eyes (n = 4), as well as eyes of 16 deceased donors without diabetic retinopathy were subjected to immunohistochemical studies, using a panel of antibodies directed against candidate markers of complement activation. RESULTS: Extensive deposits of complement C5b-9 complexes were detected in the choriocapillaris immediately underlying the Bruch membrane and densely surrounding the capillaries, in all 50 diabetic retinopathy specimens. Complement deposition was sometimes also observed around the larger choroidal vessels. Similarly intense staining for C5b-9 was absent in 25 of the 26 of the other donor eyes. Positive staining was observed in a case of systemic amyloidosis. Staining for C3d positively correlated with C5b-9 staining, corroborating the notion that complement activation had occurred in situ. Furthermore, positive staining was found for vitronectin, which forms stable complexes with extracellular C5b-9. When present, deposits under the pigment epithelium and drusen also stained positively for the activated complement components, independent of diabetic retinopathy. In contrast, there was no positive staining for C-reactive protein (CRP), mannose-binding lectin (MBL), C1q, or C4, indicating that complement activation did not occur through a C4-dependent pathway. CONCLUSIONS: The presence of C3d, C5b-9, and vitronectin indicates that complement activation occurs to completion, possibly through the alternative pathway in the choriocapillaris in eyes affected by diabetic retinopathy. Complement activation at this site may evoke a spectrum of pathologic sequelae that could contribute to ocular tissue disease and visual impairment. PMID- 11923253 TI - Corneal pathogenesis of Staphylococcus aureus strain Newman. AB - PURPOSE: To determine the pathogenic role of gamma- and alpha-toxin in a rabbit model of Staphylococcus aureus keratitis. METHODS: S. aureus strains Newman (expressing gamma-toxin), Newman Delta(hlg) (deficient in gamma-toxin), Newman Delta(hlg)/pCU1 hlg(+) (chromosomal gamma-toxin-deficient mutant rescued by a plasmid encoding gamma-toxin), and Newman Delta(hla) (alpha-toxin-deficient) were intrastromally injected into rabbit corneas. Eyes were scored by slit lamp examination (SLE), and bacterial colony-forming units (CFU) per cornea were determined at 15, 20, and 25 hours after infection. Histologic examination of corneas was performed. Rabbits were immunized against alpha-toxin and subsequently challenged with S. aureus strain Newman. Western blot analyses of culture supernatants were performed to detect alpha-toxin production. RESULTS: All strains grew equivalently, producing approximately 7 log CFU per cornea at 25 hours after infection. SLE scores at 20 and 25 hours after infection revealed that strains Newman Delta(hlg) and Newman Delta(hla), although virulent, caused significantly less ocular damage and inflammation than their parent or the gamma toxin genetically rescued strain (P ET-1 > CCh; their effects on PGE(2) release were in the following order: PGF(2alpha) > CCh > ET-1; and their effects on inositol phosphate production were in the following order: CCh > ET-1 > PGF(2alpha). Both the basal- and stimulated release of AA were significantly higher in the g-HCM cells than in the n-HCM cells (for PGF(2alpha), 60% vs. 151%). The basal release of PGE(2) in g-HCM cells was two- to fivefold higher than that of n-HCM cells, and there are alterations in the effects of the agonists on PGE(2) release. Agonist-induced inositol phosphate production in g-HCM cells was considerably lower than that of n-HCM cells (CCh, 58% vs. 421%), and the amount of PLC-beta(1) expressed in g-HCM cells, compared with that in n-HCM cells, was markedly reduced (by 44%). CONCLUSIONS: These data are the first to show that basal and agonist induced AA release and inositol phosphate production as well as expression of PLC beta(1) are altered in g-HCM cells compared with that of n-HCM cells. The molecular mechanisms underlying these alterations in g-HCM cells could include changes in sensitivity and number of receptors, overexpression of PLA(2) and the cyclooxygenases, and underexpression of PLC-beta(1). Alterations in these signaling pathways in g-HCM cells could contribute to changes in the uveoscleral outflow pathway, which may lead to an increase in IOP in patients with glaucoma. Comparative studies on the signaling pathways in g- and n-HCM cells can provide important information about the regulation of uveoscleral outflow and the pathologic course of glaucoma. PMID- 11923257 TI - Role of syndecan-1 in leukocyte-endothelial interactions in the ocular vasculature. AB - PURPOSE: Leukocyte endothelial interactions are a key feature of ocular angiogenesis but also play a role in nonproliferative vascular alterations as are found in early diabetes or uveitis. The adhesion of leukocytes to endothelial cells during inflammation is a multistep process that involves leukocyte rolling, adhesion, and extravasation mediated by selectins, cell adhesion molecules (CAMs), integrins, and chemokines. Heparan sulfate (HS) is known to bind to and modify the function of these molecules under physiological conditions. In this study, the role of the HS proteoglycan syndecan-1 in mediating leukocyte endothelial interactions in the ocular vasculature was investigated. METHODS: Mice carrying a deletion in the gene encoding the cell surface HS proteoglycan syndecan-1 (sdc1) were used to study the interactions of leukocytes and endothelial cells in vivo, using a perfusion technique with FITC-coupled ConA and intravital microscopy. RESULTS: In a retina perfusion model, Sdc1(-/-) mice showed increased leukocyte adhesion that was largely attributable to the leukocytes. Intravital microscopy studies revealed a dramatic increase in adhesion after tumor necrosis factor (TNF)-alpha treatment of sdc1(-/-) mice compared with similarly treated wild-type mice. The higher degree of leukocyte adhesion may account for the increase in inflammation-mediated corneal angiogenesis observed in sdc1(-/-) mice. CONCLUSIONS: The results indicate a role for syndecan-1 as a negative regulator of leukocyte-mediated inflammatory responses. Thus, syndecan-1 could have use as a target for prevention of pathologic leukocyte-endothelial interactions in angiogenesis and inflammation. PMID- 11923258 TI - Effects of topical antiglaucoma eye drops on prostaglandin E(2)-induced aqueous flare elevation in pigmented rabbits. AB - PURPOSE: To evaluate the role of topical instillation of some antiglaucoma agents on experimental elevation of aqueous flare induced by prostaglandin E(2) (PGE(2)) in pigmented rabbits. METHODS: Transcorneal diffusion of PGE(2) (25 microg/mL or 7.09 x 10(-2) mM) with the use of a glass cylinder was achieved to produce aqueous flare elevation in pigmented rabbits. An antiglaucoma agent was topically administered before application of PGE(2). Aqueous flare was measured with a laser flare cell meter. RESULTS: A single instillation of apraclonidine 1.15%, two instillations of epinephrine 1.25%, two instillations of dipivefrin 0.1%, and two instillations and one instillation of dipivefrin 0.04% eye drops inhibited 98%, 96%, 87%, 73%, and 47% of PGE(2)-induced aqueous flare elevation, respectively. Timolol 0.5%, nipradilol 0.25%, dorzolamide 1%, and pilocarpine 2% eye drops had no effects on the increase of PGE(2)-induced flare. CONCLUSIONS: Apraclonidine, epinephrine, and dipivefrin eye drops inhibit PGE(2)-induced elevation of aqueous flare in pigmented rabbits. PMID- 11923259 TI - Enhancement of HCO(3)(-) permeability across the apical membrane of bovine corneal endothelium by multiple signaling pathways. AB - PURPOSE: In this study, the involvement of signaling pathways in the regulation of HCO(3)(-) permeability across the apical membrane of the corneal endothelium was examined. METHODS: Cultured bovine corneal endothelial cells (CBCECs) were grown to confluence on permeable membranes. Apical and basolateral sides were perfused with a HCO(3)(-)-rich Cl(-)-free Ringer's solution (28.5 mM; pH 7.5). Relative changes in apical HCO(3)(-) permeability were assayed by pulsing the apical perfusion bath with a low-HCO(3)(-) Cl(-)-free Ringer's solution (2.85 mM; pH 6.5), in the presence or absence of agonists or inhibitors, and comparing the rates of change in intracellular pH (pH(i)), as measured with a pH-sensitive dye. Ca(2+)-activated signaling was measured with the Ca(2+)-sensitive dye Fura-2. Qualitative changes in membrane potential (E(m)) were measured with a voltage sensitive dye. RT-PCR using calcium-activated chloride channel (CLCA)-specific primers was used to examine the expression of CLCA in the corneal endothelium. RESULTS: The adenoceptor agonist adenosine (20 M) enhanced HCO(3)(-) permeability by a factor of 2. Forskolin (40 microM) exerted a 6.3-fold increase of HCO(3)(-) permeability, which was inhibited by the Cl(-) channel blockers, glibenclamide (50 microM) and niflumic acid (100 microM). Adenosine triphosphate (ATP) and ATPgammaS, P(2) receptor agonists that increased intracellular Ca(2+) in corneal endothelium, enhanced HCO(3)(-) permeability by 87% and 79%, respectively. ATPgammaS induced depolarization of the E(m), consistent with anion channel activation, rather than activation of Ca(2+)-dependent K(+) channels, which could secondarily increase extrusion of anions by E(m) hyperpolarization. Cyclopiazonic acid (CPA), an endoplasmic reticulum (ER) Ca(2+)-pump inhibitor that increased [Ca(2+)](i), also enhanced HCO(3)(-) permeability by 95%. Both the calmodulin kinase II (CaMKII) inhibitor KN-62 and the PKC inhibitor bisindolylmaleimide I (BIMI), decreased HCO(3)(-) permeability induced by ATPgammaS. The PKC activator PMA also increased HCO(3)(-) permeability by a factor of 1.8. RT-PCR using CLCA specific primers showed the expression of CLCA1 in both fresh and cultured BCECs. CONCLUSIONS: Activation of adenoceptors and purinoceptors enhances HCO(3)(-) permeability across the apical membrane of the cultured corneal endothelium. Multiple signaling pathways (PKA, PKC, and Ca(2+)/CaMKII) contribute to the HCO(3)(-) transport in cultured corneal endothelium. Both cAMP and Ca(2+) activated Cl(-) channels (possibly CLCA) may be involved in HCO(3)(-) transport. PMID- 11923260 TI - Net glutathione secretion across primary cultured rabbit conjunctival epithelial cell layers. AB - PURPOSE: Metabolism and transport of glutathione (GSH), the endogenous thiol antioxidant, in conjunctival tissue to date are poorly understood. The purpose of the present study was to define transport characteristics of GSH in primary cultured rabbit conjunctival epithelial cells (RCECs). METHODS: RCECs were grown on membrane filters to exhibit tight barrier properties (transepithelial electrical resistance, TEER, approximately 1 k(Omega)/cm(2)). Uptake, efflux, and transepithelial transport of GSH were determined in the presence or absence of extracellular Na(+) under conditions of inhibition of GSH biosynthesis and degradation. Uptake was determined at 15 minutes after instillation of (3)H-GSH to the apical or basolateral bathing fluid. GSH efflux was estimated from the time course of release of prebiosynthesized (35)S-GSH. Transepithelial transport was assessed by instillation of (3)H-GSH in either the apical or basolateral bathing fluid, followed by sampling from respective contralateral sides. RESULTS: Apical uptake and efflux showed Na(+) dependency up to 65%. GSH uptake in the initial 15 minutes was linear in the presence of 1 mM GSH (labeled and unlabeled) in Na(+)-containing buffer. The uptake rate was higher from the apical fluid than from the basolateral fluid. A Hill analysis of the Na(+)-dependent process yielded a coupling ratio for Na(+) to GSH of 1.25:1. The efflux rate of GSH into the apical fluid was marginally dependent on the apical presence of Na(+) and was significantly greater than that in the basolateral fluid. Basolateral efflux of GSH was primarily Na(+) independent, whereas basolateral uptake almost exclusively was Na(+) dependent. Depolarizing the RCEC membrane potential decreased GSH efflux into either apical or basolateral fluids (5 pmol/min small middle dot 10(6) cells). Hyperpolarization significantly increased the rate of GSH efflux into the apical fluid (120 pmol/min small middle dot 10(6) cells), whereas the basolateral efflux was not affected. Apparent permeability of GSH across RCEC layers was approximately eight times higher in the basolateral-to apical (secretion) direction than the opposite (absorption) direction. CONCLUSIONS: GSH is transported across RCEC membranes by both Na(+)-dependent and -independent processes. Analysis of the Na(+)-dependent uptake process gave an approximate 1:1 coupling ratio for Na(+)-GSH cotransport. The Na(+)-independent component is highly sensitive to cell membrane potential. Net secretion of GSH into the apical fluid may play a role in the protection of conjunctival tissue and tear film from oxidant insults. PMID- 11923261 TI - Cytoprotective effect of thioredoxin against retinal photic injury in mice. AB - PURPOSE: To determine the protective role of thioredoxin (TRX), an endogenous redox (reduction and oxidation) regulator, against retinal photic injury in mice. METHODS: Four-week-old BALB/c mice were exposed to white fluorescent light (8000 lux) for 2 hours. The number of both the photoreceptor cell nuclei and the TUNEL positive photoreceptor cell nuclei were counted to determine the severity of damage. Expression of endogenous TRX was analyzed in the retinal samples by immunohistochemistry and Western blot. Recombinant (r)TRX or mutant rTRX, in which cysteines in the active site are replaced with serines, was injected intravitreously into BALB/c mice before light exposure. Oxidized and tyrosine phosphorylated proteins were analyzed in retinal samples to examine the antioxidative effect of TRX. The number of photoreceptor cell nuclei and the DNA ladder in the retinal samples were analyzed. RESULTS: A significant reduction was observed in the number of photoreceptor cells and induction of TUNEL-positive nuclei after light exposure. TRX expression was enhanced in both the neural retina and retinal pigment epithelium after light exposure. The amounts of oxidized and tyrosine-phosphorylated proteins decreased in the neural retinas of the rTRX-treated mice compared with the vehicle- or mutant rTRX-treated mice. The reduction of photoreceptor cells and formation of a DNA ladder were suppressed by rTRX pretreatment but not with mutant rTRX. CONCLUSIONS: TRX is induced in the retinal tissue after light exposure. Intraocular injection of rTRX suppresses photo-oxidative stress. TRX intensification may be a useful therapeutic strategy to prevent retinal photic injury. PMID- 11923262 TI - Expression of pigment epithelium-derived factor in normal adult rat eye and experimental choroidal neovascularization. AB - PURPOSE: Pigment epithelium-derived factor (PEDF) is a protein produced by the retinal pigment epithelial (RPE) cells. Recent studies have implicated PEDF in activities that are inhibitory to angiogenesis. In this study, the expression of PEDF was investigated in normal rat eyes and in eyes with experimentally induced choroidal neovascularization and compared with the expression of vascular endothelial growth factor (VEGF). METHODS: Choroidal neovascularization was induced by laser photocoagulation in rat eyes. At intervals of up to 2 weeks after photocoagulation, the eyes were removed and prepared for in situ hybridization and immunohistochemical study. In situ hybridization was performed with digoxigenin-labeled PEDF riboprobes. Protein expression of PEDF and VEGF was studied immunohistochemically. RESULTS: In normal adult rat eyes, PEDF mRNA was observed mainly in the corneal epithelial and endothelial cells, lens epithelial cells, ciliary epithelial cells, retinal ganglion cells, and the RPE cells. During the development of choroidal neovascularization, PEDF mRNA, PEDF protein, and VEGF protein were strongly detected in many cells within the laser lesions at 3 days after photocoagulation, after which levels gradually declined. However, PEDF was still expressed in the RPE cells that proliferated and covered the neovascular tissues at 2 weeks, whereas VEGF protein was weakly expressed in endothelial cells in choroidal neovascularization. CONCLUSIONS: PEDF is expressed in different cell types of normal rat eyes. The expression of PEDF was detected in the choroidal neovascular tissues induced by photocoagulation, and these findings suggest that PEDF may modulate the process of choroidal neovascularization. PMID- 11923264 TI - An in vitro study of ceftazidime and vancomycin concentrations in various fluid media: implications for use in treating endophthalmitis. AB - PURPOSE: To investigate the precipitation process of a mixture of vancomycin and ceftazidime by equilibrium dialysis and determine its subsequent effect on the level of free antibiotics for treatment of endophthalmitis. METHODS: Concentrations of vancomycin and ceftazidime in an equilibrium dialysis chamber were measured during the equilibrium process by high-performance liquid chromatography. Normal saline (NS), balanced salt solution (BSS), and vitreous were used separately as the medium of dialysis. RESULTS: Precipitation of ceftazidime occurred at 37 degrees C but not at room temperature and did not affect the pH of the medium. It formed precipitate on its own or when mixed with vancomycin in all the three media of NS, BSS, and vitreous. More precipitation was formed if ceftazidime was initially prepared in BSS than in NS. After 168 hours in the dialysis chambers, ceftazidime prepared in NS precipitated to 54% of that in vitreous, compared with 88% if prepared in BSS. At 48 hours, ceftazidime prepared in NS decreased from an initial concentration of 137.5 to 73.4 microg/mL in vitreous medium and to 6.3 microg/mL if prepared in BSS. Precipitation of vancomycin was negligible. CONCLUSIONS: Based on this in vitro investigation, ceftazidime precipitates in vitreous at body temperature, regardless of the presence of vancomycin. NS is preferred to BSS as a preparation medium for antibiotics for intravitreal injection, because the extent of ceftazidime precipitation is less. However, due to precipitation, the concentration of free ceftazidime in vitreous may not be sufficiently high for antibacterial activity against most common organisms. PMID- 11923263 TI - Functional magnetic resonance imaging of the retina. AB - PURPOSE: This study explored the feasibility of mapping the retina's responses to visual stimuli noninvasively, by using functional magnetic resonance imaging (fMRI). METHODS: fMRI was performed on a 9.4-Tesla scanner to map activity-evoked signal changes of the retina-choroid complex associated with visual stimulation in anesthetized cats (n = 6). Three to 12 1-mm slices were acquired in a single shot using inversion-recovery, echo-planar imaging with a nominal in-plane resolution of 468 x 468 microm(2). Visual stimuli were presented to the full visual field and to the upper and lower visual fields. The stimuli were drifting or stationary gratings, which were compared with the dark condition. Activation maps were computed using cross-correlation analysis and overlaid on anatomic images. Multislice activation maps were reconstructed and flattened onto a two dimensional surface. RESULTS: fMRI activation maps showed robust increased activity in the retina-choroid complex after visual stimulation. The average stimulus-evoked fMRI signal increase associated with drifting-grating stimulus was 1.7% +/- 0.5% (P < 10(-4), n = 6) compared with dark. Multislice functional images of the retina flattened onto a two-dimensional surface showed relatively uniform activation. No statistically significant activation was observed in and around the optic nerve head. Hemifield stimulation studies demonstrated that stimuli presented to the upper half of the visual field activated the lower part of the retina, and stimuli presented to the lower half of the visual field activated the upper part of the retina, as expected. Signal changes evoked by the stationary gratings compared with the dark basal condition were positive but were approximately half that evoked by the drifting gratings (1.0% +/- 0.1% versus 2.1% +/- 0.3%, P < 10(-4)). CONCLUSIONS: To the best of our knowledge, this is the first fMRI study of the retina, demonstrating its feasibility in imaging retinal function dynamically in a noninvasive manner and at relatively high spatial resolution. PMID- 11923265 TI - Nature of the cone ON-pathway dysfunction in melanoma-associated retinopathy. AB - PURPOSE: To establish the basis for an ON-pathway abnormality of the cone system in melanoma-associated retinopathy (MAR) through analysis of the electroretinogram (ERG) and visual evoked potential (VEP). METHODS: Two patients with MAR syndrome whose sera produced immunolabeling of retinal bipolar cells participated in the study. Full-field ERGs were recorded in response to brief flashes, to rapid-on and rapid-off sawtooth stimuli at a temporal frequency of 8 Hz, and to sine-wave stimuli at temporal frequencies ranging from 8 to 96 Hz. Fundamental responses to the sine-wave stimuli were evaluated within the context of a vector-summation model of the depolarizing bipolar cell (DBC) and hyperpolarizing bipolar cell (HBC) contributions to the response fundamental. VEPs were recorded to the onset of luminance increments and decrements that had contrasts of 10%, 20%, and 50%. The patients' results were compared with those of age-similar control subjects. RESULTS: The patients with MAR showed abnormal ERG responses to luminance increments, consisting of a marked attenuation of the initial portion of the b-wave, but their ERG responses to luminance decrements were normal in amplitude and timing. The ERG temporal response functions of the patients with MAR had normal amplitudes at frequencies of 32 Hz and higher, with a constant phase lag across these frequencies, but larger-than-normal amplitudes at frequencies below 32 Hz, and a phase lead at 8 Hz. Their VEP responses showed a marked delay to increments but only a minimal delay to decrements. CONCLUSIONS: Within the context of the vector-summation model, the ERG findings in the patients with MAR are more consistent with an attenuation of the DBC contribution to the ERG response than with a DBC response delay. The delayed VEP responses of the patients with MAR to luminance increments may represent a late response of the OFF system to increment onset. PMID- 11923266 TI - Correlation between rod photoreceptor numbers and levels of ocular pigmentation. AB - PURPOSE: Ocular melanin synthesis modulates rod photoreceptor production, because in albino eyes, rod numbers are reduced by approximately 30%. In this study, rod numbers and ocular rhodopsin concentrations were measured in intermediate pigmentation phenotypes to determine whether proportional reductions in melanin are correlated with proportional changes in rod numbers. Further, patterns of cell production and death were examined around the time of birth, when rod production peaks, to determine whether there are abnormalities in these features associated with hypopigmentation. METHODS: Four mouse pigmentation phenotypes were used: fully pigmented, albino, Beige, and Himalayan. The latter two are intermediate-pigmentation phenotypes, with Beige having markedly more pigment than Himalayan. Ocular melanin concentrations were measured during development and at maturity. Rods were counted at maturity and measurements of ocular rhodopsin undertaken. Mitotic and pyknotic cells were also counted in neonates. RESULTS: Rods and ocular rhodopsin were reduced in both Beige and Himalayan mice below levels found in fully pigmented mice, but not to levels found in albino animals. This was more marked in Himalayan than Beige mice, reflecting the lower concentration of melanin found in the former compared with the latter, both in development and at maturity. Although patterns of cell production were elevated in the hypopigmented animals, such patterns varied. CONCLUSIONS: Rod numbers are modulated within a range between that in fully pigmented and albino phenotypes by the concentration of ocular melanin. However, in these animals, there is no obvious correlation between these events and patterns of cell production and death in neonates. PMID- 11923267 TI - Inflammatory response after scatter laser photocoagulation in nonphotocoagulated retina. AB - PURPOSE: Macular edema is one of the most serious adverse effects after retinal scatter laser photocoagulation. It has been suggested that the changes in the distribution of retinal blood flow or the inflammatory reaction after photocoagulation may be involved in the pathogenesis of macular edema, but little information is available about its exact mechanism. This study was designed to evaluate quantitatively leukocyte-endothelial cell interactions and vascular permeability in the nonphotocoagulated portions of the retina after partial scatter laser photocoagulation. METHODS: Argon laser photocoagulation was performed in one half of the retina in male pigmented rats (n = 90). In the other half of the retina, leukocyte dynamics after photocoagulation were evaluated in vivo with acridine orange digital fluorography. Retinal vessel permeability was quantified by using Evans blue dye. RESULTS: Scatter laser photocoagulation caused significant inflammatory leukocyte-endothelial interactions not only in the photocoagulated but also in the untreated half of the retina. In the nonphotocoagulated half of the retina, the number of leukocytes rolling along the major retinal veins increased after photocoagulation and peaked at 12 hours (14.3 +/- 4.5 cells/min per vessel). Leukocyte accumulation in the untreated half of the retina increased after photocoagulation, with a peak of 47.5 +/- 13.0 cells/mm(2) 24 hours after photocoagulation. Retinal vascular permeability in the untreated half of the retina gradually increased after photocoagulation. CONCLUSIONS: Scatter laser photocoagulation increased leukocyte rolling and subsequent accumulation in both the photocoagulated and the untreated portions of the retina. The accumulated leukocytes may be involved in the augmented vascular permeability in the untreated retina, resulting in retinal edema after photocoagulation. PMID- 11923268 TI - Effect of dietary zeaxanthin on tissue distribution of zeaxanthin and lutein in quail. AB - PURPOSE: The xanthophyll carotenoids (lutein and zeaxanthin) are hypothesized to delay progression of age-related macular degeneration. The quail has a cone dominant retina that accumulates carotenoids. The purpose of these experiments was to characterize the carotenoid composition of retina, serum, liver, and fat in quail and to determine whether dietary enrichment with zeaxanthin alters zeaxanthin or lutein concentrations in these tissues. METHODS: Quail were fed for 6 months with a commercial turkey diet (T group; n = 8), carotenoid-deficient diet (C- group; n = 8), or a carotenoid-deficient diet supplemented with 35 mg 3R,3'R-zeaxanthin per kilogram of food, (Z+ group; n = 8). Zeaxanthin was derived from Sphingobacterium multivorum (basonym Flavobacterium). Carotenoids in serum, retina, liver, and fat were analyzed by HPLC. RESULTS: As in the primate fovea, the retina accumulated zeaxanthin, lutein, and cryptoxanthin, and preferentially absorbed zeaxanthin (P < 0.005). In contrast, lutein was preferentially absorbed by liver (P < 0.01) and fat (P < 0.0001). In supplemented females, zeaxanthin increased approximately 4-fold in retina, and 74-, 63- and 22-fold in serum, liver, and fat, respectively. In males, zeaxanthin was elevated approximately 3 fold in retina, and 42-, 17-, and 12-fold in serum, liver, and fat, respectively. Birds fed the Z+ diet absorbed a higher fraction of dietary lutein into serum, but lutein was reduced in the retina (P < 0.05). CONCLUSIONS: Xanthophyll profiles in quail mimic those in primates. Dietary supplements of zeaxanthin effectively increased zeaxanthin concentrations in serum, retina, liver, and fat. The robust response to zeaxanthin supplementation identifies the quail as an animal model for exploration of factors regulating delivery of dietary carotenoids to the retina. PMID- 11923269 TI - Involvement of oxidative mechanisms in blue-light-induced damage to A2E-laden RPE. AB - PURPOSE: The lipofuscin fluorophore A2E is known to be an initiator of blue-light induced apoptosis in retinal pigment epithelial cells (RPE). The purpose of this study was to evaluate the role of oxidative mechanisms in mediating the cellular damage. METHODS: Human RPE (ARPE-19) cells that had accumulated A2E were exposed to blue light in the presence and absence of oxygen, and nonviable cells were quantified. Potential suppressors (histidine, azide, 1,4-diazabicyclooctane [DABCO], and 1,3-dimethyl-2-thiourea [DMTU]) and enhancers (deuterium oxide [D(2)O] and 3-aminotriazole [3-AT]) of oxidative damage, were also screened for their ability to modulate the frequency of nonviable cells. A2E in PBS, with and without an oxygen-depleter or singlet-oxygen quencher and A2E-laden RPE, were exposed to 430-nm light and examined by reversed-phase high performance liquid chromatography (HPLC) and fast atom bombardment mass spectrometry (FAB-MS). RESULTS: The death of blue-light-illuminated A2E-laden RPE was blocked in oxygen depleted media. When A2E-laden RPE were transferred to D(2)O-based media and then irradiated (480 nm), the number of nonviable cells was increased, whereas the latter was decreased in the presence of histidine, DABCO, and azide. Conversely, no affect was observed with 3-AT and DMTU. When A2E, in either acellular or cellular environments, was irradiated at 430 nm, FAB-MS revealed the generation of a series of higher molecular mass derivatives of A2E. The sizes of these species increased by increments of mass 16. The generation of these photo products was accompanied by the consumption of A2E, the latter being diminished, however, when illumination was performed after oxygen depletion and in the presence of a singlet-oxygen quencher. CONCLUSIONS: The augmentation of cell death in the presence of D(2)O and the protection afforded by quenchers and scavengers of singlet oxygen, indicates that the generation of singlet oxygen may be involved in the mechanisms leading to the death of A2E-containing RPE cells after blue light illumination. The finding that irradiation also produces oxygen dependent photochemical changes in A2E, indicates that the effects of singlet oxygen may be mediated either directly or through the generation of reactive photo-products of A2E. PMID- 11923270 TI - Polyamine-dependent migration of retinal pigment epithelial cells. AB - PURPOSE: Migration of retinal pigment epithelial (RPE) cells can be triggered by disruption of the RPE monolayer or injury to the neural retina. Migrating cells may re-establish a confluent monolayer, or they may invade the neural retina and disrupt visual function. The purpose of this study was to examine the role of endogenous polyamines in mechanisms of RPE migration. METHODS: Endogenous polyamine levels were determined in an immortalized RPE cell line, D407, using HPLC. Activities of the two rate-limiting enzymes for polyamine synthesis, ornithine decarboxylase (ODC), and S-adenosylmethionine decarboxylase (SAMdc), were measured by liberation of ((14)CO(2))(.) Migration was assessed in confluent cultures by determining the number of cells migrating into a mechanically denuded area. All measurements were obtained both in control cultures and in cultures treated with synthesis inhibitors that deplete endogenous polyamines. Subcellular localization of endogenous polyamines was determined using a polyamine antibody. RESULTS: The polyamines, spermidine and spermine, as well as their precursor, putrescine, were normal constituents of RPE cells. The two rate-limiting synthetic enzymes were also present, and their activities were stimulated dramatically by addition of serum to the culture medium. Cell migration was similarly stimulated by serum exposure. When endogenous polyamines were depleted, migration was blocked. When polyamines were replenished through uptake, migration was restored. Polyamine immunoreactivity was limited to membrane patches in quiescent cells. In actively migrating and dividing cells, immunoreactivity was enhanced throughout the cytoplasm. CONCLUSIONS: Polyamines are essential for RPE migration. Pharmacologic manipulation of the polyamine pathway could provide a therapeutic strategy for regulating anomalous migration. PMID- 11923271 TI - A survey of molecular expression by photoreceptors after experimental retinal detachment. AB - PURPOSE: To describe changes in the localization patterns and levels of rod and cone photoreceptor proteins after experimental retinal detachment (RD). METHODS: Cat retinas were detached for 1, 3, 7, or 28 days, at which time the eyecups were placed in fixative for immunocytochemical analysis or homogenized for biochemistry. Immunocytochemistry was performed using 19 probes for molecules known to be associated with photoreceptors. Protein concentrations were determined using enzyme-linked immunosorbent assay or Western blot analysis. Cone cell death was analyzed by double labeling with TdT-dUTP terminal nick-end labeling and cone-specific antibodies. RESULTS: Although some cones died, many survived long-term RD. Although their profiles may have changed, rod photoreceptors continued to express most of the molecules studied as long as they were alive. In contrast, the cones failed to label with almost all probes specific to them after 3 to 7 days of detachment. The exception was phosducin, which localized to both rods and cones and, in 28-day detachments, increased to 180% of the amount in normal retina. CONCLUSIONS: Rods and cones respond differently to RD. This difference may account for a faster return of rod vision and for the lingering changes in color vision and acuity that are often reported after successful reattachment surgeries. PMID- 11923272 TI - Alterations of slow and fast rod ERG signals in patients with molecularly confirmed Stargardt disease type 1. AB - PURPOSE: To investigate the slow and fast rod signals of the scotopic 15-Hz flicker ERG in patients with molecularly confirmed Stargardt disease type I (STGD1). There is evidence that these slow and the fast rod ERG signals can be attributed to the rod bipolar-AII cell pathway and the rod-cone coupling pathway, respectively. METHODS: Twenty-seven patients with STGD1 with mutations in both alleles of the ABCA4 gene were included. Scotopic ERG response amplitudes and phases to flicker intensities ranging from -3.37 to -0.57 log scotopic troland x sec (log scot td x sec) were measured at a flicker frequency of 15 Hz. In addition, scotopic standard ERGs were obtained. Twenty-two normal subjects served as controls. RESULTS: The amplitudes of both the slow and fast rod ERG signals were significantly reduced in the STGD1 group. The phases of the slow rod signals lagged significantly, whereas those of the fast rod signals did not. The standard scotopic ERG did not reveal significant alterations. CONCLUSIONS: The results provide evidence that a defective ABCA4 transporter can functionally affect both the rod bipolar-AII cell pathway and the rod-cone coupling pathway. In STGD1, the scotopic 15-Hz flicker ERG may reveal subtle abnormalities at different sites within the rod system that remain undetected by standard ERG techniques. PMID- 11923273 TI - Effects of haloperidol on K(+) currents in acutely isolated rat retinal ganglion cells. AB - PURPOSE: Effects of haloperidol on K(+) currents (IKs) of rat retinal ganglion cells (RGCs) were examined, with the hypothesis that its alteration of IKs explains alterations in the pattern electroretinogram (PERG). METHODS: Fast blue was injected into superior colliculi of rats (3-8 days old) to identify RGCs under epifluorescence illumination after retrograde transport to retinas. Retinas were dissected, treated enzymatically, and dissociated with trituration. Effects of haloperidol on membrane currents at -70 mV, voltage-dependent IK, and Ca(2+) dependent K(+) currents (K(Ca)) were examined by whole-cell patch voltage clamp. Na(+) currents were abolished by tetrodotoxin (1 microM; TTX). Voltage-gated IKs were isolated by Ca(2+)-free perfusate. Persistent and transient components of the voltage-sensitive IKs were isolated by prepulses, and sensitivity of each component to tetraethylammonium (TEA, 20 mM) and 4-aminopyridine (5 mM) was tested. K(Ca) was identified by its response to TEA, charybdotoxin (CTX), and apamin. Haloperidol (0.01-100 microM) was instilled into the perfusate dissolved in dimethyl sulfoxide (DMSO). RESULTS: Currents recorded at -70 mV were not affected by haloperidol, whereas the persistent component of the voltage dependent IK was reversibly reduced by haloperidol, with a dose dependence fitted with the Hill equation (median inhibitory concentration [IC(50)] = 4.2 microM). The transient component of the voltage-gated IK was less sensitive to haloperidol. Haloperidol (10 nM) blocked the apamin-sensitive K(Ca) but not the CTX-sensitive K(Ca). CONCLUSIONS: Haloperidol reduced voltage-dependent IKs in RGCs, but at a higher concentration than that needed to antagonize dopamine receptors. Haloperidol (10 nM) blocked the apamin-sensitive K(Ca) which modulates the firing rate of RGCs and may contribute to the alteration of PERG. PMID- 11923274 TI - Effects of the duration of early strabismus on the binocular responses of neurons in the monkey visual cortex (V1). AB - PURPOSE: To determine how the duration of early strabismus influences the severity of loss of disparity sensitivity in V1 neurons and the effects of extensive poststrabismus visual experience on the maintenance of functional binocular connections. METHODS: Concomitant strabismus was optically simulated in 10 rhesus monkeys using a prism-rearing procedure. The onset of strabismus was kept constant at 4 weeks of age and the duration was maintained for 2, 4, or 8 weeks. In one group of monkeys (infants), the neurophysiological experiments were conducted immediately after the period of rearing with prisms. In another group (adults), after the termination of the prism-rearing regimen at either 8 or 12 weeks of age, the monkeys were kept in a normal housing environment until maturity and behavioral testing was conducted before the recording experiments to determine the animal's monocular and binocular visual capacities. To assess the effects of the period of early strabismus on binocular interactions in V1, extracellular single-unit recording methods were used in anesthetized and paralyzed monkeys, and dichoptic sine-wave gratings were used as stimuli. RESULTS: In all strabismic monkeys, the sensitivity of V1 units to interocular spatial phase disparity (disparity sensitivity) was significantly reduced, and the prevalence of binocular suppression was higher than that found in age-matched control animals. Although 8 weeks of strabismus resulted in a slightly larger loss of disparity sensitivity, the overall effects of the duration of strabismus were surprisingly small in infant strabismic monkeys. After poststrabismus visual experience, a small but significantly higher degree of disparity sensitivity was noted in V1 if prism-rearing was terminated after 4 weeks of strabismus (i.e., at 8 weeks of age), but not after 8 weeks of strabismus (i.e., at 12 weeks of age). CONCLUSIONS: A brief period (2 weeks) of misalignment after the emergence of stereopsis is sufficient to drastically reduce the functional binocular connections in V1, and longer periods of strabismus result in little additional loss in disparity sensitivity. Clinically, these results suggest that taking corrective measures for infantile esotropes before the known onset age for stereopsis may be important for maintaining better binocular sensory function and better interocular alignment at later stages of development. PMID- 11923276 TI - Sensitivity and specificity of frequency doubling perimetry in neuro-ophthalmic disorders: a comparison with conventional automated perimetry. AB - PURPOSE: Frequency-doubling technology (FDT) perimetry was developed as a screening test for glaucoma. Patients with damage to the neuro-ophthalmic sensory visual pathways have different patterns of visual loss than patients with glaucoma. The current study was designed to determine the sensitivity and specificity of FDT as a screening test, compared with conventional automated perimetry (CAP) in neuro-ophthalmic disorders and to test the extent to which it may isolate the M(y) cells. METHODS: FDT and CAP were performed in 97 patients with sensory neuro-ophthalmic disorders and 42 subjects from the general population. The total and pattern-deviation probability plots for test loci common to the two perimetric tests were compared. The gold standard was an unequivocal clinical diagnosis. RESULTS: The sensitivity of FDT was 81.3%, with a specificity of 76.2%. The difference in sensitivity and specificity of CAP, 87.5% and 81.0%, respectively, was not statistically significant (by chi(2) test). In subjects with optic neuropathies, the similarity of the defect shown on FDT and CAP was judged good or fair in 62 of 72 cases. The extent of the defect as seen with FDT and CAP was equal in 41 of 72 cases, more extensive with FDT in 12, and more extensive with CAP in 19. In the patients with hemianopia, scattered abnormal test locations with FDT testing masked the hemianopic nature of the defect in 15 of 25 patients. Also, test locations along the vertical midline in densely hemianopic areas were seen with FDT testing in some patients with hemianopia, probably due to light scatter across the vertical midline and into the uninvolved hemianopic field. CONCLUSIONS: FDT has sensitivity and specificity similar to that of CAP for detecting visual field defects in patients with optic neuropathies. However, defects in patients with hemianopias may be missed because of the presence of scattered abnormal test locations and failure to detect test locations along the vertical meridian. The defects demonstrated by both tests in patients with optic neuropathies are similar in number, extent, and shape of the defects. This suggests FDT may not be isolating the magnocellular (M) cells with nonlinear responses to stimulus contrast (M(y) cells) in patients with visual loss. PMID- 11923275 TI - The effect of letter spacing on reading speed in central and peripheral vision. AB - PURPOSE: Crowding, the adverse spatial interaction due to proximity of adjacent letters, has been suggested as an explanation for slow reading in peripheral vision. The purpose of this study was to examine whether reading speed can be improved in normal peripheral vision by increasing the letter spacing. Also tested was whether letter spacing imposes a different limit on reading speed of small versus large print. METHODS: Six normal observers read aloud single, short sentences presented on a computer monitor, one word at a time, by rapid serial visual presentation (RSVP). Reading speeds were calculated based on the RSVP exposure durations yielding 80% correctly read words. Letters were rendered in Courier, a fixed-width font. Testing was conducted at the fovea, 5 degrees and 10 degrees in the inferior visual field. The critical print size (CPS) was first determined for each observer by measuring reading speeds for four print sizes, using the standard letter spacing (center-to-center separation of adjacent letters; standard Courier spacing: 1.16 times the width of the lowercase x). Text was then presented at 0.8 x or 1.5x CPS, and reading speed was measured for five letter spacings, ranging from 0.5 times to 2 times the standard spacing. RESULTS: As expected, reading speed was highest at the fovea, decreased with eccentricity, and was faster for the larger print size. At all eccentricities and for both print sizes, reading speed increased with letter spacing, up to a critical letter spacing, and then either remained constant at the same reading speed or decreased slightly for larger letter spacings. The value of the critical letter spacing was very close to the standard letter spacing and did not depend on eccentricity or print size. CONCLUSIONS: Increased letter spacing beyond the standard size, which presumably decreases the adverse effect of crowding, does not lead to an increase in reading speed in central or peripheral vision. PMID- 11923277 TI - Engineering-enhanced protein secretory expression in yeast with application to insulin. AB - Adaptation to efficient heterologous expression is a prerequisite for recombinant proteins to fulfill their clinical and biotechnological potential. We describe a rational strategy to optimize the secretion efficiency in yeast of an insulin precursor by structure-based engineering of the folding stability. The yield of a fast-acting insulin analogue (Asp(B28)) expressed in yeast was enhanced 5-fold by engineering a specific interaction between an aromatic amino acid in the connecting peptide and a phenol binding site in the hydrophobic core of the molecule. This insulin precursor is characterized by significantly enhanced folding stability. The improved folding properties enhanced the secretion efficiency of the insulin precursor from 10 to 50%. The precursor remains fully in vitro convertible to mature fast-acting insulin. PMID- 11923278 TI - Absence of metabolic cross-correction in Tay-Sachs cells: implications for gene therapy. AB - We have investigated the ability of a receptor-mediated gene transfer strategy (cross-correction) to restore ganglioside metabolism in fibroblasts from Tay Sachs (TS) patients in vitro. TS disease is a GM2 gangliosidosis attributed to the deficiency of the lysosomal enzyme beta-hexosaminidase A (HexA) (beta-N acetylhexosaminidase, EC ). The hypothesis is that transduced cells overexpressing and secreting large amounts of the enzyme would lead to a measurable activity in defective cells via a secretion-recapture mechanism. We transduced NIH3T3 murine fibroblasts with the LalphaHexTN retroviral vector carrying the cDNA encoding for the human Hex alpha-subunit. The Hex activity in the medium from transduced cells was approximately 10-fold higher (up to 75 milliunits) than observed in non-transduced cells. TS cells were cultured for 72 h in the presence of the cell medium derived from the transduced NIH3T3 cells, and they were analyzed for the presence and catalytic activity of the enzyme. Although TS cells were able to efficiently uptake a large amount of the soluble enzyme, the enzyme failed to reach the lysosomes in a sufficient quantity to hydrolyze the GM2 ganglioside to GM3 ganglioside. Thus, our results showed that delivery of the therapeutic HexA was not sufficient to correct the phenotype of TS cells. PMID- 11923279 TI - Cell surface targeting and clustering interactions between heterologously expressed PSD-95 and the Shal voltage-gated potassium channel, Kv4.2. AB - Kv4.2 is a voltage-gated potassium channel that is critical in controlling the excitability of myocytes and neurons. Processes that influence trafficking and surface distribution patterns of Kv4.2 will affect its ability to contribute to cellular functions. The scaffolding/clustering protein PSD-95 regulates trafficking and distribution of several receptors and Shaker family Kv channels. We therefore investigated whether the C-terminal valine-serine-alanine-leucine (VSAL) of Kv4.2 is a novel binding motif for PSD-95. By using co immunoprecipitation assays, we determined that full-length Kv4.2 and PSD-95 interact when co-expressed in mammalian cell lines. Mutation analysis in this heterologous expression system showed that the VSAL motif of Kv4.2 is necessary for PSD-95 binding. PSD-95 increased the surface expression of Kv4.2 protein and caused it to cluster, as shown by deconvolution microscopy and biotinylation assays. Deleting the C-terminal VSAL motif of Kv4.2 eliminated these effects, as did substituting a palmitoylation-deficient PSD-95 mutant. In addition to these effects of PSD-95 on Kv4.2 distribution, the channel itself promoted redistribution of PSD-95 to the cell surface in the heterologous expression system. This work represents the first evidence that a member of the Shal subfamily of Kv channels can bind to PSD-95, with functional consequences. PMID- 11923280 TI - Akt enhances Mdm2-mediated ubiquitination and degradation of p53. AB - p53 plays a key role in DNA damage-induced apoptosis. Recent studies have reported that the phosphatidylinositol 3-OH-kinase-Akt pathway inhibits p53 mediated transcription and apoptosis, although the underlying mechanisms have yet to be determined. Mdm2, a ubiquitin ligase for p53, plays a central role in regulation of the stability of p53 and serves as a good substrate for Akt. In this study, we find that expression of Akt reduces the protein levels of p53, at least in part by enhancing the degradation of p53. Both Akt expression and serum treatment induced phosphorylation of Mdm2 at Ser186. Akt-mediated phosphorylation of Mdm2 at Ser186 had little effect on the subcellular localization of Mdm2. However, both Akt expression and serum treatment increased Mdm2 ubiquitination of p53. The serum-induced increase in p53 ubiquitination was blocked by LY294002, a phosphatidylinositol 3-OH-kinase inhibitor. Moreover, when Ser186 was replaced by Ala, Mdm2 became resistant to Akt enhancement of p53 ubiquitination and degradation. Collectively, these results suggest that Akt enhances the ubiquitination-promoting function of Mdm2 by phosphorylation of Ser186, which results in reduction of p53 protein. This study may shed light on the mechanisms by which Akt promotes survival, proliferation, and tumorigenesis. PMID- 11923281 TI - TLR4 and MD-2 expression is regulated by immune-mediated signals in human intestinal epithelial cells. AB - The normal intestinal epithelium is not inflamed despite contact with a high density of commensal bacteria. Intestinal epithelial cells (IEC) express low levels of TLR4 and MD-2 and are lipopolysaccharide (LPS)-unresponsive. We hypothesized that immune-mediated signals regulate the expression of TLR4 and MD 2 in IEC. Expression of TLR4 and MD-2 was examined in normal colonic epithelial cells or intestinal epithelial cell lines. The effect of the cytokines interferon (IFN)-gamma, IFN-alpha, and tumor necrosis factor-alpha (TNF-alpha) on TLR4 and MD-2 expression was examined by reverse transcription-PCR and Western blot. NF kappaB transcriptional activation and interleukin-8 secretion were used as measures of LPS responsiveness. Native colonic epithelial cells and IEC lines express a low level of TLR4 and MD-2 mRNA. IFN-gamma regulates MD-2 expression in both IEC lines, whereas IFN-gamma and TNF-alpha regulate TLR4 mRNA expression in IEC lines. Pre-incubation with IFN-gamma and/or TNF-alpha sensitizes IEC to LPS dependent interleukin-8 secretion. To examine MD-2 transcriptional regulation, we cloned a 1-kb sequence proximal to the MD-2 gene translational start site. This promoter directed expression of a reporter gene in endothelial cells and IEC. IFN gamma positively regulated MD-2 promoter activity in IEC. Co-expression of a STAT inhibitor, SOCS3, blocked IFN-gamma-mediated MD-2 promoter activation. T cell derived cytokines lead to increased expression of TLR4 and MD-2 and LPS-dependent pro-inflammatory cytokine secretion in IEC. IFN-gamma regulates expression of the critical TLR4 co-receptor MD-2 through the Janus tyrosine kinase-STAT pathway. Th1 cytokines may initiate or perpetuate intestinal inflammation by altering toll like receptor expression and bacterial reactivity. PMID- 11923282 TI - Regulation of metallothionein transcription by the metal-responsive transcription factor MTF-1: identification of signal transduction cascades that control metal inducible transcription. AB - Every living organism must detoxify nonessential metals and carefully control the intracellular concentration of essential metals. Metallothioneins, which are small, cysteine-rich, metal-binding proteins, play an important role in these processes. In addition, the transcription of their cognate genes is activated in response to metal exposure. The zinc finger transcription factor MTF-1 plays a central role in the metal-inducible transcriptional activation of metallothionein and other genes involved in metal homeostasis and cellular stress response. Here we report that the phosphorylation of MTF-1 plays a critical role in its activation by zinc and cadmium. Inhibitor studies indicate that multiple kinases and signal transduction cascades, including those mediated by protein kinase C, tyrosine kinase, and casein kinase II, are essential for zinc- and cadmium inducible transcriptional activation. In addition, calcium signaling is also involved in regulating metal-activated transcription. In contrast, cAMP-dependent protein kinase may not be directly involved in the metal response. Contrary to what has been reported for other transcription factors, inhibition of transcriptional activation does not impair the binding of MTF-1 to DNA, suggesting that phosphorylation is not regulating DNA binding. Elevated phosphorylation of MTF-1 is observed under condition of protein kinase C inhibition, suggesting that specific dephosphorylation of this transcription factor contributes to its activation. PMID- 11923284 TI - Crystal structures of deoxy- and carbonmonoxyhemoglobin F1 from the hagfish Eptatretus burgeri. AB - Hagfish are extremely primitive jawless fish of disputed ancestry. Although generally classed with lampreys as cyclostomes ("round mouths"), it is clear that they diverged from them several hundred million years ago. The crystal structures of the deoxy and CO forms of hemoglobin from a hagfish (Eptatretus burgeri) have been solved at 1.6 and 2.1 A, respectively. The deoxy crystal contains one dimer and two monomers in a unit cell, with the dimer being similar to that found in lamprey deoxy-Hb, but with a larger interface and different relative orientation of the partner chains. Ile(E11) and Gln(E7) obstruct ligand binding in the deoxy form and make room for ligands in the CO form, but no interaction path between the two hemes could be identified. The BGH core structure, which forms the alpha1beta1 interface of all vertebrate alpha2beta2 tetrameric Hbs, is conserved in hagfish and lamprey Hbs. It was shown previously that human and cartilaginous fish Hbs have independently evolved stereochemical mechanisms other than the movement of the proximal histidine to regulate ligand binding at the hemes. Our results therefore suggest that the formation of the alpha2beta2 tetramer using the BGH core and the mechanism of quaternary structure change evolved between the branching points of hagfish and lampreys from other vertebrates. PMID- 11923283 TI - CheW binding interactions with CheA and Tar. Importance for chemotaxis signaling in Escherichia coli. AB - The initial signaling events underlying the chemotactic response of Escherichia coli to aspartic acid occur within a ternary complex that includes Tar (an aspartate receptor), CheA (a protein kinase), and CheW. Because CheW can bind to CheA and to Tar, it is thought to serve as an adapter protein in this complex. The functional importance of CheW binding interactions, however, has not been investigated. To better define the role of CheW and its binding interactions, we performed biochemical characterization of six mutant variants of CheW. We examined the ability of the purified mutant CheW proteins to bind to CheA and Tar, to promote formation of active ternary complexes, and to support chemotaxis in vivo. Our results indicate that mutations which eliminate CheW binding to Tar (V36M) or to CheA (G57D) result in a complete inability to form active ternary complexes in vitro and render the CheW protein incapable of mediating chemotaxis in vivo. The in vivo signaling pathway can, however, tolerate moderate changes in CheW-Tar and CheW-CheA affinities observed with several of the mutants (G133E, G41D, and 154ocr). One mutant (R62H) provided surprising results that may indicate a role for CheW in addition to binding CheA/receptors and promoting ternary complex formation. PMID- 11923285 TI - Increased expression of Hsp40 chaperones, transcriptional factors, and ribosomal protein Rpp0 can cure yeast prions. AB - The Sup35 (eRF3) translation termination factor of Saccharomyces cerevisiae can undergo a prion-like conformational conversion, thus resulting in the [PSI(+)] nonsense-suppressor determinant. In vivo this process depends critically on the chaperone Hsp104, whose lack or overexpression can cure [PSI(+)]. The use of artificial prion [PSI(+)PS] based on a hybrid Sup35PS with prion domain from the yeast Pichia methanolica allowed us to uncover three more chaperones, Ssb1, Ssa1, and Ydj1, whose overexpression can cure prion determinants. Here, we used the [PSI(+)PS] to search a multicopy yeast genomic library for novel factors able to cure prions. It was found that overexpression of the Hsp40 family chaperones Sis1 and Ynl077w, chaperone Sti1, transcriptional factors Sfl1 and Ssn8, and acidic ribosomal protein Rpp0 can interfere with propagation and manifestation of [PSI(+)PS] in a prion strain-specific manner. Some of these factors also affected the manifestation and propagation of conventional [PSI(+)]. Excess of Sfl1, Ssn8, and Rpp0 influenced at least one of the tested chaperone-specific promoters, SSA4, HSP104, and model promoters, with either the heat shock or stress response elements. Thus, the induction of chaperone expression by these proteins could explain their prion-curing effects. PMID- 11923286 TI - Multilayer formation upon compression of surfactant monolayers depends on protein concentration as well as lipid composition. An atomic force microscopy study. AB - The determinants for the formation of multilayers upon compression of surfactant monolayers were investigated by compressing films, beyond the squeeze-out plateau, to a surface tension of 22 millinewtons/m. Atomic force microscopy was used to visualize the topography of lipid films containing varying amounts of native surfactant protein B (SP-B). These films were compared with films containing synthetic peptides based on the N terminus of human SP-B: monomeric mSP-B-(1-25) or dimeric dSP-B-(1-25). The formation of typical hexagonal network structures as well as the height of protrusions were shown to depend on the concentration of SP-B. Protrusions of bilayer height were formed from physiologically relevant concentrations of 0.2-0.4 mol % (4.5-8.5 wt %) SP-B upwards. Much higher concentrations of SP-B-(1-25) peptides were needed to obtain network structures, and protrusion heights were not equal to those found for films with native SP-B. A striking observation was that while protrusions formed in films of 1,2-dipalmitoyl-sn-glycero-3-phosphocholine (DPPC)/1,2-dipalmitoyl-sn glycero-3-(phospho-rac-(1-glycerol)) (DPPG) (80/20) had single bilayer thickness, those formed in DPPC/1-palmitoyl-2-oleoyl-sn-glycero-3-(phospho-rac-(1-glycerol)) (80/20) had various heights of multilayers, whereas those seen in DPPC/1 palmitoyl-2-oleoyl-sn-glycero-3-phosphocholine/DPPG (60/20/20) were mainly of bilayer height. For the first time direct observations by atomic force microscopy show (i) that a certain minimal concentration of SP-B is required for the formation of layered protrusions upon film compression, (ii) that protrusion height depends on whether the phospholipids contain an unsaturated fatty acyl chain, and (iii) that protrusion height also depends on whether the unsaturated acyl chain is present in phosphatidylcholine or in phosphatidylglycerol. PMID- 11923287 TI - Characterization of type II phosphatidylinositol 4-kinase isoforms reveals association of the enzymes with endosomal vesicular compartments. AB - Phosphorylation of phosphatidylinositol (PI) to PI 4-phosphate is one of the key reactions in the production of phosphoinositides, lipid regulators of several cellular functions. This reaction is catalyzed by multiple enzymes that belong either to the type II or the type III family of PI 4-kinases. Type III enzymes are structurally similar to PI 3-kinases and are sensitive to PI 3-kinase inhibitors. In contrast, the recent cloning of the first type II PI 4-kinase enzyme defined a novel enzyme family. Here we characterize a new member of this family, the type IIbeta enzyme that has been identified in the NCBI data base based on its homology to the first-cloned type IIalpha enzyme. The type IIbeta enzyme has a primary transcript size of approximately 3.8 kb and shows wide tissue distribution. It contains an open reading frame of 1.4 kb, encoding a protein of approximately 54 kDa. Sequence comparison reveals a high degree of similarity to the type IIalpha enzyme within the C-terminal catalytic domain but significantly lower homology within the N-terminal region. Expression of both enzyme yields increased PI 4-kinase activity that is associated with the microsomal membrane fractions and is significantly lower for the type IIbeta than the type IIalpha form. Both enzymes use PI as their primary substrate and have no detectable activity on PI monophosphates. Epitope-tagged as well as green fluorescent protein-tagged forms of both enzymes localize primarily to intracellular membranes and show prominent co-localization with early endosomes and recycling endosomes but not with the Golgi. These compartments participate in the processing of both the transferrin receptor and the G protein-coupled AT(1A) angiotensin receptor. Our data indicate the existence of multiple forms of type II PI 4-kinase in mammalian cells and suggest that their functions are related to the endocytic pathway. PMID- 11923288 TI - Characterization of a family of IAA-amino acid conjugate hydrolases from Arabidopsis. AB - The mechanisms by which plants regulate levels of the phytohormone indole-3 acetic acid (IAA) are complex and not fully understood. One level of regulation appears to be the synthesis and hydrolysis of IAA conjugates, which function in both the permanent inactivation and temporary storage of auxin. Similar to free IAA, certain IAA-amino acid conjugates inhibit root elongation. We have tested the ability of 19 IAA-l-amino acid conjugates to inhibit Arabidopsis seedling root growth. We have also determined the ability of purified glutathione S transferase (GST) fusions of four Arabidopsis IAA-amino acid hydrolases (ILR1, IAR3, ILL1, and ILL2) to release free IAA by cleaving these conjugates. Each hydrolase cleaves a subset of IAA-amino acid conjugates in vitro, and GST-ILR1, GST-IAR3, and GST-ILL2 have K(m) values that suggest physiological relevance. In vivo inhibition of root elongation correlates with in vitro hydrolysis rates for each conjugate, suggesting that the identified hydrolases generate the bioactivity of the conjugates. PMID- 11923289 TI - Activation of Maf/AP-1 repressor Bach2 by oxidative stress promotes apoptosis and its interaction with promyelocytic leukemia nuclear bodies. AB - The oxidative stress response operates by inducing the expression of genes that counteract the stress. We show here that the oxidative stress-responsive transcription factor Bach2 is a generic inhibitor of gene expression directed by the 12-O-tetradecanoylphorbol-13-acetate response element, the Maf recognition element, and the antioxidant-responsive element. The Bach2-enhanced green fluorescent protein bicistronic retrovirus was used to monitor the fate of Bach2 expressing cells at the single cell level. Bach2 exerted an inhibitory effect on NIH3T3 cell proliferation and caused massive apoptosis upon mild oxidative stress in both NIH3T3 and Raji B-lymphoid cells. Interestingly, Bach1, a highly homologous protein, could not induce cell death, demonstrating the specificity for the apoptosis induction. Although both oxidative stress and leptomycin B, an inhibitor of nuclear export, induce nuclear accumulation of Bach2, the leptomycin B-induced nuclear accumulation of Bach2 was not sufficient to elicit apoptosis. Upon oxidative stress, Bach2 formed nuclear foci that associated with promyelocytic leukemia nuclear bodies. Our results suggest that Bach2 constitutes a cell lineage-specific system that couples oxidative stress and cell death and that inhibition of 12-O-tetradecanoylphorbol-13-acetate response element, the Maf recognition element, and the antioxidant-responsive element upon oxidative stress may be critical determinants for apoptosis. PMID- 11923290 TI - Targeted expression of a protease-resistant IGFBP-4 mutant in smooth muscle of transgenic mice results in IGFBP-4 stabilization and smooth muscle hypotrophy. AB - The insulin-like growth factor-binding protein 4 (IGFBP-4), the most abundant IGF binding protein produced by rodent smooth muscle cells (SMC), is degraded by specific protease(s) potentially releasing IGF-I for local bioactivity. IGFBP-4 protease(s) recognizes basic residues within the midregion of the molecule. We constructed a mutant IGFBP-4 with the cleavage domain substitution 119 KHMAKVRDRSKMK-133 to 119-AAMAAVADASAMA-133. Myc-tagged native and IGFBP-4.7A retained equivalent IGF-I binding affinity. Whereas native IGFBP-4 was cleaved by SMC-conditioned medium, IGFBP-4.7A was completely resistant to proteolysis. To explore the function of the protease-resistant IGFBP-4 in vivo, expression of the mutant and native proteins was targeted to SMC of transgenic mice by means of a smooth muscle alpha-actin promoter. Transgene expression was confined to SMC-rich tissues in all lines. Bladder and aortic immunoreactive IGFBP-4/transgene mRNA ratios in SMP8-BP4.7A mice were increased by 2- to 4-fold relative to SMP8-BP4 mice, indicating that the IGFBP-4.7A protein was stabilized in vivo. SMP8-BP4.7A mice had lower aortic, bladder, and stomach weight and intestinal length relative to SMP8-BP4 counterparts matched for protein expression by Western blotting. Thus, IGFBP-4.7A results in greater growth inhibition than equivalent levels of native IGFBP-4 in vivo, demonstrating a role for IGFBP-4 proteolysis in the regulation of IGF-I action. PMID- 11923291 TI - Thrombospondin stimulates focal adhesion disassembly through Gi- and phosphoinositide 3-kinase-dependent ERK activation. AB - The matricellular protein thrombospondin (TSP) stimulates stress fiber and focal adhesion disassembly through a sequence (hep I) in its heparin-binding domain. TSP/hep I signals focal adhesion disassembly by binding cell surface calreticulin (CRT) and activating phosphoinositide 3-kinase (PI3K). However, other components of this signaling pathway have not been identified. We now show that TSP induces focal adhesion disassembly through activation of pertussis toxin (PTX)-sensitive G proteins and ERK phosphorylation. PTX pretreatment inhibits TSP/hep I-mediated focal adhesion disassembly as well as PI3K activation. In addition, membrane permeable Galpha(i2)- and Gbetagamma-blocking peptides inhibit hep I-mediated focal adhesion disassembly. Hep I stimulates a transient increase in ERK activation, which is abrogated by both PTX and PI3K inhibitors. Inhibiting ERK activation with MEK inhibitors blocks hep I-mediated focal adhesion disassembly, indicating that ERK activation is required for cytoskeletal reorganization. G protein signals and ERK phosphorylation are induced by TSP binding to cell surface CRT, because CRT null mouse embryonic fibroblasts (MEF) fail to stimulate ERK phosphorylation in response to TSP/hep I treatment. These data show that G(i) protein and ERK, in concert with PI3K, are stimulated by TSP.CRT interactions at the cell surface to induce de-adhesive changes in the cytoskeleton. PMID- 11923292 TI - ATP hydrolysis by mammalian RAD51 has a key role during homology-directed DNA repair. AB - Disruption of the gene encoding RAD51, the protein that catalyzes strand exchange during homologous recombination, leads to the accumulation of chromosome breaks and lethality in vertebrate cells. As RAD51 is implicated in BRCA1- and BRCA2 mediated tumor suppression as well as cellular viability, we have begun a functional analysis of a defined RAD51 mutation in mammalian cells. By using a dominant negative approach, we generated a mouse embryonic stem cell line that expresses an ATP hydrolysis-defective RAD51 protein, hRAD51-K133R, at comparable levels to the endogenous wild-type RAD51 protein, whose expression is retained in these cells. We found that these cells have increased sensitivity to the DNA damaging agents mitomycin C and ionizing radiation and also exhibit a decreased rate of spontaneous sister-chromatid exchange. By using a reporter for the repair of a single chromosomal double-strand break, we also found that expression of the hRAD51-K133R protein specifically inhibits homology-directed double-strand break repair. Furthermore, expression of a BRC repeat from BRCA2, a peptide inhibitor of an early step necessary for strand exchange, exacerbates the inhibition of homology-directed repair in the hRAD51-K133R expressing cell line. Thus, ATP hydrolysis by RAD51 has a key role in various types of DNA repair in mammalian cells. PMID- 11923293 TI - The central domain of Escherichia coli TyrR is responsible for hexamerization associated with tyrosine-mediated repression of gene expression. AB - TyrR from Escherichia coli regulates the expression of genes for aromatic amino acid uptake and biosynthesis. Its central ATP-hydrolyzing domain is similar to conserved domains of bacterial regulatory proteins that interact with RNA polymerase holoenzyme associated with the alternative sigma factor, sigma(54). It is also related to the common module of the AAA+ superfamily of proteins that is involved in a wide range of cellular activities. We expressed and purified two TyrR central domain polypeptides. The fragment comprising residues 188-467, called TyrR-(188-467), was soluble and stable, in contrast to that corresponding to the conserved core from residues 193 to 433. TyrR-(188-467) bound ATP and rhodamine-ATP with association constants 2- to 5-fold lower than TyrR and hydrolyzed ATP at five times the rate of TyrR. In contrast to TyrR, which is predominantly dimeric at protein concentrations less than 10 microm in the absence of ligands, or in the presence of ATP or tyrosine alone, TyrR-(188-467) is a monomer, even at high protein concentrations. Tyrosine in the presence of ATP or ATPgammaS promotes the oligomerization of TyrR-(188-467) to a hexamer. Tyrosine-dependent repression of gene transcription by TyrR therefore depends on ligand binding and hexamerization determinants located in the central domain polypeptide TyrR-(188-467). PMID- 11923295 TI - Lysolipids do not inhibit influenza virus fusion by interaction with hemagglutinin. AB - The interaction of a spin-labeled lysophosphatidylcholine analog with intact and bromelain-treated influenza viruses as well as with the bromelain-solubilized hemagglutinin ectodomain has been studied. The inhibition of fusion of influenza viruses with erythrocytes by the lysophosphatidylcholine analog was similar to that observed for non-labeled lysophosphatidylcholine. Only a weak interaction of the lysophosphatidylcholine analog with the hemagglutinin ectodomain was observed even upon triggering the conformational change of the ectodomain at a low pH. A significant interaction of spin-labeled lysophosphatidylcholine with the hemagglutinin ectodomain of intact viruses was observed neither at neutral nor at low pH, whereas a strong interaction of the lipid analog with the viral lipid bilayer was evident. We suggest that the high number of lipid binding sites of the virus bilayer and their affinity compete efficiently with binding sites of the hemagglutinin ectodomain. We conclude that the inhibition of influenza virus fusion by lysolipids is not mediated by binding to the hemagglutinin ectodomain, preventing its interaction with the target membrane. The results unambiguously argue for an inhibition mechanism based on the action of lysolipid inserted into the lipid bilayer. PMID- 11923294 TI - 5-Hydroxytryptamine 4(a) receptor is coupled to the Galpha subunit of heterotrimeric G13 protein. AB - Serotonin (5-hydroxytryptamine (5-HT)) is an important neurotransmitter that regulates multiple events in the central nervous system. Many of the 5-HT functions are mediated via G protein-coupled receptors that are coupled to multiple heterotrimeric G proteins, including G(s), G(i), and G(q) subfamilies (Martin, G. R., Eglen, R. M., Hamblin, M. W., Hoyer, D., and Yocca, F. (1998) Trends Pharmacol. Sci. 19, 2-4). Here we show for the first time that the 5 hydroxytryptamine 4(a) receptor (5-HT(4(a))) is coupled not only to heterotrimeric G(s) but also to G(13) protein, as assessed both by biochemical and functional assays. Using reconstitution of 5-HT(4(a)) receptor with different G proteins in Spodoptera frugiperda (Sf.9) cells, we have proved that agonist stimulation of receptor-induced guanosine 5'-(3-O-thio)triphosphate binding to Galpha(13) protein. We then determined that expression of 5-HT(4(a)) receptor in mammalian cells induced constitutive- as well as agonist-promoted activation of a transcription factor, serum response element, through the activation of Galpha(13) and RhoA. Finally, we have determined that expression of 5-HT(4(a)) receptor in neuroblastoma x glioma NIE-115 cells cause RhoA-dependent neurite retraction and cell rounding under basal conditions and after agonist stimulation. These data suggest that by activating 5-HT(4(a)) receptor-G(13) pathway, serotonin plays a prominent role in regulating neuronal architecture in addition to its classical role in neurotransmission. PMID- 11923297 TI - Characterization of oligopeptides that cross-react with carbohydrate-specific antibodies by real time kinetics, in-solution competition enzyme-linked immunosorbent assay, and immunological analyses. AB - Phage displaying random cyclic 7-mer, and linear 7-mer and 12-mer peptides at the N terminus of the coat protein, pIII, were panned with the murine monoclonal antibody, 9-2-L379 specific for meningococcal lipo-oligosaccharide. Five cyclic peptides with two sequence motifs, six linear 7-mers, and five linear 12-mers with different sequence motifs were identified. Only phage displaying cyclic peptides were specifically captured by and were antigenic for 9-2-L379. Monoclonal antibody 9-2-L379 exhibited "apparent" binding affinities to the cyclic peptides between 11 and 184 nm, comparable with lipo-oligosaccharide. All cyclic peptides competed with the binding of 9-2-L379 to lipo-oligosaccharide with EC(50) values in the range 10-105 microm, which correlated with their apparent binding affinities. Structural modifications of the cyclic peptides eliminated their ability to bind and compete with monoclonal antibody 9-2-L379. Mice (C3H/HeN) immunized with the cyclic peptide with optimal apparent binding affinity and EC(50) of competition elicited cross-reactive antibodies to meningococcal lipo-oligosaccharide with end point dilution serum antibody titers of 3200. Cyclic peptides were converted to T-cell-dependent immunogens without disrupting these properties by C-terminal biotinylation and complexing with NeutrAvidin. The data indicate that constrained peptides can cross-react with a carbohydrate-specific antibody with greater specificity than linear peptides, and critical to this specificity is their structural conformation. PMID- 11923298 TI - Transcriptional program of mouse osteoclast differentiation governed by the macrophage colony-stimulating factor and the ligand for the receptor activator of NFkappa B. AB - Cytokines macrophage colony stimulating factor (M-CSF) and the receptor activator of NFkappaB ligand (RANKL) induce differentiation of bone marrow hematopoietic precursor cells into bone-resorbing osteoclasts without the requirement for stromal cells of mesenchymal origin. We used this recently described mouse cell system and oligonucleotide microarrays representing about 9,400 different genes to analyze gene expression in hematopoietic cells undergoing differentiation to osteoclasts. The ability of microarrays to detect the genes of interest was validated by showing expression and expected regulation of several osteoclast marker genes. In total 750 known transcripts were up-regulated by > or =2-fold, and 91% of them at an early time in culture, suggesting that almost the whole differentiation program is defined already in pre-osteoclasts. As expected, M-CSF alone induced the receptor for RANKL (RANK), but also, unexpectedly, other RANK/NFkappaB pathway components (TRAF2A, PI3-kinase, MEKK3, RIPK1), providing a molecular explanation for the synergy of M-CSF and RANKL. Furthermore, interleukins, interferons, and their receptors (IL-1alpha, IL-18, IFN-beta, IL 11Ralpha2, IL-6/11R gp130, IFNgammaR) were induced by M-CSF. Although interleukins are thought to regulate osteoclasts via modulation of M-CSF and RANKL expression in stromal cells, we showed that a mix of IL-1, IL-6, and IL-11 directly increased the activity of osteoclasts by 8.5-fold. RANKL induced about 70 novel target genes, including chemokines and growth factors (RANTES (regulated on activation, normal T cell expressed and secreted), PDGFalpha, IGF1), histamine, and alpha1A-adrenergic receptors, and three waves of distinct receptors, transcription factors, and signaling molecules. In conclusion, M-CSF induced genes necessary for a direct response to RANKL and interleukins, while RANKL directed a three-stage differentiation program and induced genes for interaction with osteoblasts and immune and nerve cells. Thus, global gene expression suggests a more dynamic role of osteoclasts in bone physiology than previously anticipated. PMID- 11923300 TI - Insulin-like growth factor-binding protein-5 (IGFBP-5) stimulates growth and IGF I secretion in human intestinal smooth muscle by Ras-dependent activation of p38 MAP kinase and Erk1/2 pathways. AB - Insulin-like growth factor-binding protein-5 (IGFBP-5) and insulin-like growth factor-I (IGF-I) are produced by human intestinal smooth muscle cells. Endogenous IGF-I stimulates growth and increases IGFBP-5 secretion. IGFBP-5 augments the effects of IGF-I by facilitating interaction of IGF-I with the IGF-I receptor tyrosine kinase. Andress (Andress, D. L. (1998) Am. J. Physiol. 274, E744-E750) and Berfield et al. (Berfield, A. K., Andress, D. L., and Abrass, C. K. (2000) Kidney Int. 57, 1991-2003) have shown that in osteoblasts and kidney mesangial cells, IGFBP-5 stimulates proliferation and filopodia formation independently of IGF-I, presumably by activating a distinct IGFBP-5 receptor serine kinase. The present study determined whether IGFBP-5 exerts direct effects on growth in human intestinal smooth muscle cells and identified the intracellular signaling pathways involved. IGFBP-5 caused a concentration-dependent increase in [(3)H]thymidine incorporation and an increase in IGF-I secretion that occurred independently of IGF-I and the IGF-I receptor tyrosine kinase. IGFBP-5-induced phosphorylation of p38 MAP kinase, which was abolished by SB203580, or expression of a dominant negative Ras mutant, Ras(S17N), and phosphorylation of Erk1/2, which was abolished by a Raf1 kinase inhibitor, U1026, or expression of Ras(S17N). IGFBP-5-stimulated [(3)H]thymidine incorporation and IGF-I secretion were partly inhibited by SB203580 or U1026 and abolished by the combination of the two inhibitors or by expression of Ras(S17N). These data show that IGFBP-5 stimulates growth and IGF-I secretion in human intestinal smooth muscle cells by activation of p38 MAP kinase-dependent and Erk1/2-dependent pathways that are independent of IGF-I. A positive feedback mechanism therefore links IGFBP-5 and IGF-I secretion that reinforces their individual effects on growth. PMID- 11923299 TI - Kinetic and structural basis of reactivity of pentaerythritol tetranitrate reductase with NADPH, 2-cyclohexenone, nitroesters, and nitroaromatic explosives. AB - The reaction of pentaerythritol tetranitrate reductase with reducing and oxidizing substrates has been studied by stopped-flow spectrophotometry, redox potentiometry, and X-ray crystallography. We show in the reductive half-reaction of pentaerythritol tetranitrate (PETN) reductase that NADPH binds to form an enzyme-NADPH charge transfer intermediate prior to hydride transfer from the nicotinamide coenzyme to FMN. In the oxidative half-reaction, the two-electron reduced enzyme reacts with several substrates including nitroester explosives (glycerol trinitrate and PETN), nitroaromatic explosives (trinitrotoluene (TNT) and picric acid), and alpha,beta-unsaturated carbonyl compounds (2 cyclohexenone). Oxidation of the flavin by the nitroaromatic substrate TNT is kinetically indistinguishable from formation of its hydride-Meisenheimer complex, consistent with a mechanism involving direct nucleophilic attack by hydride from the flavin N5 atom at the electron-deficient aromatic nucleus of the substrate. The crystal structures of complexes of the oxidized enzyme bound to picric acid and TNT are consistent with direct hydride transfer from the reduced flavin to nitroaromatic substrates. The mode of binding the inhibitor 2,4-dinitrophenol (2,4-DNP) is similar to that observed with picric acid and TNT. In this position, however, the aromatic nucleus is not activated for hydride transfer from the flavin N5 atom, thus accounting for the lack of reactivity with 2,4-DNP. Our work with PETN reductase establishes further a close relationship to the Old Yellow Enzyme family of proteins but at the same time highlights important differences compared with the reactivity of Old Yellow Enzyme. Our studies provide a structural and mechanistic rationale for the ability of PETN reductase to react with the nitroaromatic explosive compounds TNT and picric acid and for the inhibition of enzyme activity with 2,4-DNP. PMID- 11923301 TI - A point mutation that confers constitutive activity to CXCR4 reveals that T140 is an inverse agonist and that AMD3100 and ALX40-4C are weak partial agonists. AB - CXCR4 is a G protein-coupled receptor for stromal-derived factor 1 (SDF-1) that plays a critical role in leukocyte trafficking, metastasis of mammary carcinoma, and human immunodeficiency virus type-1 infection. To elucidate the mechanism for CXCR4 activation, a constitutively active mutant (CAM) was derived by coupling the receptor to the pheromone response pathway in yeast. Conversion of Asn-119 to Ser or Ala, but not Asp or Lys, conferred autonomous CXCR4 signaling in yeast and mammalian cells. SDF-1 induced signaling in variants with substitution of Asn-119 to Ser, Ala, or Asp, but not Lys. These variants had similar cell surface expression and binding affinity for SDF-1. CXCR4-CAMs were constitutively phosphorylated and present in cytosolic inclusions. Analysis of antagonists revealed that exposure to AMD3100 or ALX40-4C induced G protein activation by CXCR4 wild type, which was greater in the CAM, whereas T140 decreased autonomous signaling. The affinity of AMD3100 and ALX40-4C binding to CAMs was less than to wild type, providing evidence of a conformational shift. These results illustrate the importance of transmembrane helix 3 in CXCR4 signaling. Insight into the mechanism for CXCR4 antagonists will allow for the development of a new generation of agents that lack partial agonist activity that may induce toxicities, as observed for AMD3100. PMID- 11923302 TI - Mks1p is required for negative regulation of retrograde gene expression in Saccharomyces cerevisiae but does not affect nitrogen catabolite repression sensitive gene expression. AB - The Tor1/2p signal transduction pathway regulates nitrogen catabolite repression (NCR)-sensitive (GAP1, GAT1, DAL5) and retrograde (CIT2, DLD3, IDH1/2) gene expression by controlling intracellular localization of the transcription activators, Gln3p and Gat1p, and Rtg1p and Rtg3p, respectively. The accepted pathway for this regulation is NH(3) or excess nitrogen dash, vertical Mks1p dash, vertical Ure2p dash, vertical Gln3p --> DAL5, and rapamycin or limiting nitrogen dash, vertical Torp --> Tap42 dash, vertical Mks1p --> Rtg1/3p --> CIT2, respectively. In current models, Mks1p positively regulates both Gln3p (and DAL5 expression) and Rtg1/3p (and CIT2 expression). Here, in contrast, we show the following. (i) Mks1p is a strong negative regulator of CIT2 expression and does not effect NCR-sensitive expression of DAL5 or GAP1. (ii) Retrograde carbon and NCR-sensitive nitrogen metabolism are not linked by the quality of the nitrogen source, i.e. its ability to elicit NCR, but by the product of its catabolism, i.e. glutamate or ammonia. (iii) In some instances, we can dissociate rapamycin induced CIT2 expression from Mks1p function, i.e. rapamycin does not suppress Mks1p-mediated down-regulation of CIT2 expression. These findings suggest that currently accepted models of Tor1/2p signal transduction pathway regulation require revision. PMID- 11923303 TI - Biochemical characterization of aspartyl phosphate phosphatase interaction with a phosphorylated response regulator and its inhibition by a pentapeptide. AB - The RapA and RapB proteins are aspartyl phosphate phosphatases that specifically dephosphorylate the Spo0F approximately P intermediate response regulator of the phosphorelay signal transduction system for sporulation initiation in Bacillus subtilis. The approximately 48-kDa His-tag derivative proteins were purified by metal affinity chromatography, and their molecular and biochemical characteristics were studied. RapA and RapB were found to be dimers in solution. Enzymatic activity was strongly dependent upon maintaining reducing conditions during purification and storage. RapA phosphatase activity on Spo0F approximately P is inhibited in vivo by a pentapeptide generated from the phrA gene. Native gel assays demonstrated that the RapA dimer forms a stable complex with two molecules of Spo0F approximately P or with its PhrA pentapeptide inhibitor. The pentapeptide was shown to displace Spo0F approximately P from a preformed complex with RapA. The structural organization of Rap phosphatases in tetratricopeptide repeats provides insights on the mechanisms of RapA interaction with its substrate and its inhibitor. PMID- 11923304 TI - Cloning and characterization of methenyltetrahydrofolate synthetase from Saccharomyces cerevisiae. AB - The folate derivative 5-formyltetrahydrofolate (folinic acid; 5-CHO-THF) was discovered over 40 years ago, but its role in metabolism remains poorly understood. Only one enzyme is known that utilizes 5-CHO-THF as a substrate: 5,10 methenyltetrahydrofolate synthetase (MTHFS). A BLAST search of the yeast genome using the human MTHFS sequence revealed a 211-amino acid open reading frame (YER183c) with significant homology. The yeast enzyme was expressed in Escherichia coli, and the purified recombinant enzyme exhibited kinetics similar to previously purified MTHFS. No new phenotype was observed in strains disrupted at MTHFS or in strains additionally disrupted at the genes encoding one or both serine hydroxymethyltransferases (SHMT) or at the genes encoding one or both methylenetetrahydrofolate reductases. However, when the MTHFS gene was disrupted in a strain lacking the de novo folate biosynthesis pathway, folinic acid (5-CHO THF) could no longer support the folate requirement. We have thus named the yeast gene encoding methenyltetrahydrofolate synthetase FAU1 (folinic acid utilization). Disruption of the FAU1 gene in a strain lacking both 5 aminoimidazole-4-carboxamide ribonucleotide (AICAR) transformylase isozymes (ADE16 and ADE17) resulted in a growth deficiency that was alleviated by methionine. Genetic analysis suggested that intracellular accumulation of the purine intermediate AICAR interferes with a step in methionine biosynthesis. Intracellular levels of 5-CHO-THF were determined in yeast disrupted at FAU1 and other genes encoding folate-dependent enzymes. In fau1 disruptants, 5-CHO-THF was elevated 4-fold over wild-type yeast. In yeast lacking MTHFS along with both AICAR transformylases, 5-CHO-THF was elevated 12-fold over wild type. 5-CHO-THF was undetectable in strains lacking SHMT activity, confirming SHMT as the in vivo source of 5-CHO-THF. Taken together, these results indicate that S. cerevisiae harbors a single, nonessential, MTHFS activity. Growth phenotypes of multiply disrupted strains are consistent with a regulatory role for 5-CHO-THF in one carbon metabolism and additionally suggest a metabolic interaction between the purine and methionine pathways. PMID- 11923306 TI - Crystal structure of Lyme disease variable surface antigen VlsE of Borrelia burgdorferi. AB - VlsE is an outer surface lipoprotein of Borrelia burgdorferi that undergoes antigenic variation through an elaborate gene conversion mechanism and is thought to play a major role in the immune response to the Lyme disease borellia. The crystal structure of recombinant variant protein VlsE1 at 2.3-A resolution reveals that the six variable regions form loop structures that constitute most of the membrane distal surface of VlsE, covering the predominantly alpha-helical, invariant regions of the protein. The surface localization of the variable amino acid segments appears to protect the conserved regions from interaction with antibodies and hence may contribute to immune evasion. PMID- 11923305 TI - Mitotic activation of protein-tyrosine phosphatase alpha and regulation of its Src-mediated transforming activity by its sites of protein kinase C phosphorylation. AB - During mitosis, the catalytic activity of protein-tyrosine phosphatase (PTP) alpha is enhanced, and its inhibitory binding to Grb2, which specifically blocks Src dephosphorylation, is decreased. These effects act synergistically to activate Src in mitosis. We show here that these effects are abrogated by mutation of Ser180 and/or Ser204, the sites of protein kinase C-mediated phosphorylation within PTPalpha. Moreover, either a Ser-to-Ala substitution or serine dephosphorylation specifically eliminated the ability of PTPalpha to dephosphorylate and activate Src even during interphase. This explains why the substitutions eliminated PTPalpha transforming activity, even though PTPalpha interphase dephosphorylation of nonspecific substrates was only slightly decreased. This occurred without change in the phosphorylation of PTPalpha at Tyr789, which is required for "phosphotyrosine displacement" during Src dephosphorylation. Thus, in addition to increasing PTPalpha nonspecific catalytic activity, Ser180 and Ser204 phosphorylation (along with Tyr789 phosphorylation) regulates PTPalpha substrate specificity. This involves serine phosphorylation dependent differential modulation of the affinity of Tyr(P)789 for the Src and Grb2 SH2 domains. The results suggest that protein kinase C may participate in the mitotic activation of PTPalpha and Src and that there are intramolecular interactions between the PTPalpha C-terminal and membrane-proximal regions that are regulated, at least in part, by serine phosphorylation. PMID- 11923307 TI - Molecular characterization of the Arabidopsis thaliana flavoprotein AtHAL3a reveals the general reaction mechanism of 4'-phosphopantothenoylcysteine decarboxylases. AB - The Arabidopsis thaliana flavoprotein AtHAL3a, which is linked to plant growth and salt and osmotic tolerance, catalyzes the decarboxylation of 4' phosphopantothenoylcysteine to 4'-phosphopantetheine, a key step in coenzyme A biosynthesis. AtHAL3a is similar in sequence and structure to the LanD enzymes EpiD and MrsD, which catalyze the oxidative decarboxylation of peptidylcysteines. Therefore, we hypothesized that the decarboxylation of 4' phosphopantothenoylcysteine also occurs via an oxidatively decarboxylated intermediate containing an aminoenethiol group. A set of AtHAL3a mutants were analyzed to detect such an intermediate. By exchanging Lys(34), we found that AtHAL3a is not only able to decarboxylate 4'-phosphopantothenoylcysteine but also pantothenoylcysteine to pantothenoylcysteamine. Exchanging residues within the substrate binding clamp of AtHAL3a (for example of Gly(179)) enabled the detection of the proposed aminoenethiol intermediate when pantothenoylcysteine was used as substrate. This intermediate was characterized by its high absorbance at 260 and 280 nm, and the removal of two hydrogen atoms and one molecule of CO(2) was confirmed by ultrahigh resolution mass spectrometry. Using the mutant AtHAL3a C175S enzyme, the product pantothenoylcysteamine was not detectable; however, oxidatively decarboxylated pantothenoylcysteine could be identified. This result indicates that reduction of the aminoenethiol intermediate depends on a redox-active cysteine residue in AtHAL3a. PMID- 11923308 TI - Absence of sterol regulatory element-binding protein-1 (SREBP-1) ameliorates fatty livers but not obesity or insulin resistance in Lep(ob)/Lep(ob) mice. AB - Obesity is a common nutritional problem often associated with diabetes, insulin resistance, and fatty liver (excess fat deposition in liver). Leptin-deficient Lep(ob)/Lep(ob) mice develop obesity and those obesity-related syndromes. Increased lipogenesis in both liver and adipose tissue of these mice has been suggested. We have previously shown that the transcription factor sterol regulatory element-binding protein-1 (SREBP-1) plays a crucial role in the regulation of lipogenesis in vivo. To explore the possible involvement of SREBP-1 in the pathogenesis of obesity and its related syndromes, we generated mice deficient in both leptin and SREBP-1. In doubly mutant Lep(ob/ob) x Srebp-1(-/-) mice, fatty livers were markedly attenuated, but obesity and insulin resistance remained persistent. The mRNA levels of lipogenic enzymes such as fatty acid synthase were proportional to triglyceride accumulation in liver. In contrast, the mRNA abundance of SREBP-1 and lipogenic enzymes in the adipose tissue of Lep(ob)/Lep(ob) mice was profoundly decreased despite sustained fat, which could explain why the SREBP-1 disruption had little effect on obesity. In conclusion, SREBP-1 regulation of lipogenesis is highly involved in the development of fatty livers but does not seem to be a determinant of obesity in Lep(ob)/Lep(ob) mice. PMID- 11923309 TI - Cyclomaltodextrinase, neopullulanase, and maltogenic amylase are nearly indistinguishable from each other. AB - Over 20 enzymes denoted as cyclomaltodextrinase, maltogenic amylase, or neopullulanase that share 40-86% sequence identity with each other are found in public data bases. These enzymes are distinguished from typical alpha-amylases by containing a novel N-terminal domain and exhibiting preferential substrate specificities for cyclomaltodextrins (CDs) over starch. In this research field, a great deal of confusion exists regarding the features distinguishing the three groups of enzymes from one another. Although a different enzyme code has been assigned to each of the three different enzyme names, even a single differentiating enzymatic property has not been documented in the literature. On the other hand, an outstanding question related to this issue concerns the structural basis for the preference of these enzymes for CDs. To clarify the confusion and to address this question, we have determined the structures of two enzymes, one from alkalophilic Bacillus sp. I-5 and named cyclomaltodextrinase and the other from a Thermus species and named maltogenic amylase. The structure of the Bacillus enzyme reveals a dodecameric assembly composed of six copies of the dimer, which is the structural and functional unit of the Thermus enzyme and an enzyme named neopullulanase. The structure of the Thermus enzyme in complex with beta-CD led to the conclusion that Trp47, a well conserved N-terminal domain residue, contributes greatly to the preference for beta-CD. The common dimer formation through the novel N-terminal domain, which contributes to the preference for CDs by lining the active-site cavity, convincingly indicates that the three groups of enzymes are not different enough to preserve the different names and enzyme codes. PMID- 11923310 TI - Functional proteolytic complexes of the human mitochondrial ATP-dependent protease, hClpXP. AB - Human mitochondrial ClpP (hClpP) and ClpX (hClpX) were separately cloned, and the expressed proteins were purified. Electron microscopy confirmed that hClpP forms heptameric rings and that hClpX forms a hexameric ring. Complexes of a double heptameric ring of hClpP with hexameric hClpX rings bound on each side are stable in the presence of ATP or adenosine 5'-(3-thiotriphosphate) (ATPgammaS), indicating that a symmetry mismatch is a universal feature of Clp proteases. hClpXP displays both ATP-dependent proteolytic activity and ATP- or ATPgammaS dependent peptidase activity. hClpXP cannot degrade lambdaO protein or GFP-SsrA, specific protein substrates recognized by Escherichia coli (e) ClpXP. However, eClpX interacts with hClpP, and, when examined by electron microscopy, the resulting heterologous complexes are indistinguishable from homologous eClpXP complexes. The hybrid eClpX-hClpP complexes degrade eClpX-specific protein substrates. In contrast, eClpA can neither associate with nor activate hClpP. hClpP has an extra C-terminal extension of 28 amino acids. A mutant lacking this C-terminal extension interacts more tightly with both hClpX and eClpX and shows enhanced enzymatic activities but still does not interact with eClpA. Our results establish that human ClpX and ClpP constitute a bone fide ATP-dependent protease and confirm that substrate selection, which differs between human and E. coli ClpX, is dependent solely on the Clp ATPase. Our data also indicate that human ClpP has conserved sites required for interaction with eClpX but not eClpA, implying that the modes of interaction with ClpP may not be identical for ClpA and ClpX. PMID- 11923311 TI - Fibroblast growth factor-10 is a mitogen for urothelial cells. AB - Fibroblast growth factor (FGF)-10 plays an important role in regulating growth, differentiation, and repair of the urothelium. This process occurs through a paracrine cascade originating in the mesenchyme (lamina propria) and targeting the epithelium (urothelium). In situ hybridization analysis demonstrated that (i) fibroblasts of the human lamina propria were the cell type that synthesized FGF 10 RNA and (ii) the FGF-10 gene is located at the 5p12-p13 locus of chromosome 5. Recombinant (r) preparations of human FGF-10 were found to induce proliferation of human urothelial cells in vitro and of transitional epithelium of wild-type and FGF7-null mice in vivo. Mechanistic studies with human cells indicated two modes of FGF-10 action: (i) translocation of rFGF-10 into urothelial cell nuclei and (ii) a signaling cascade that begins with the heparin-dependent phosphorylation of tyrosine residues of surface transmembrane receptors. The normal urothelial phenotype, that of quiescence, is proposed to be typified by negligible levels of FGF-10. During proliferative phases, levels of FGF-10 rise at the urothelial cell surface and/or within urothelial cell nuclei. An understanding of how FGF-10 works in conjunction with these other processes will lead to better management of many diseases of the bladder and urinary tract. PMID- 11923313 TI - Separate analysis of twin-arginine translocation (Tat)-specific membrane binding and translocation in Escherichia coli. AB - The twin-arginine translocation (Tat) pathway exports those precursor proteins to the periplasmic space of bacteria that harbor a twin-arginine (RR) consensus motif in their signal sequences. We have reproduced translocation of several Tat substrates into inside-out plasma membrane vesicles from Escherichia coli. Translocation proceeding at an efficiency of up to 20% occurs specifically via the Tat pathway as indicated by (i) its requirement for elevated levels of the TatABC proteins in the membrane vesicles, (ii) competition by an intact twin arginine signal peptide, and (iii) susceptibility toward dissipation of the transmembrane H(+) gradient. The latter treatment, while blocking translocation, still allows for functional membrane association of Tat precursors. This is shown by the finding that translocation of isolated membrane-bound Tat precursor is restored upon re-energization of the vesicles. PMID- 11923312 TI - Complete reconstitution of the human coenzyme A biosynthetic pathway via comparative genomics. AB - The biosynthesis of CoA from pantothenic acid (vitamin B5) is an essential universal pathway in prokaryotes and eukaryotes. The CoA biosynthetic genes in bacteria have all recently been identified, but their counterparts in humans and other eukaryotes remained mostly unknown. Using comparative genomics, we have identified human genes encoding the last four enzymatic steps in CoA biosynthesis: phosphopantothenoylcysteine synthetase (EC ), phosphopantothenoylcysteine decarboxylase (EC ), phosphopantetheine adenylyltransferase (EC ), and dephospho-CoA kinase (EC ). Biological functions of these human genes were verified using a complementation system in Escherichia coli based on transposon mutagenesis. The individual human enzymes were overexpressed in E. coli and purified, and the corresponding activities were experimentally verified. In addition, the entire pathway from phosphopantothenate to CoA was successfully reconstituted in vitro using a mixture of purified recombinant enzymes. Human recombinant bifunctional phosphopantetheine adenylyltransferase/dephospho-CoA kinase was kinetically characterized. This enzyme was previously suggested as a point of CoA biosynthesis regulation, and we have observed significant differences in mRNA levels of the corresponding human gene in normal and tumor cells by Northern blot analysis. PMID- 11923314 TI - A RAC protein-binding site in the internal transcribed spacer 2 of Pre-rRNA transcripts from Schizosaccharomyces pombe. AB - The interdependence of steps in the processing of the eukaryotic preribosomal rRNA transcripts indicate that rRNA processing, at least in part, acts as a quality control mechanism to help ensure that only functional rRNA is incorporated into mature ribosomes. In search of structural components that underlie this interdependence, we have isolated a large protein complex or RAC that contains an independent binding site for all four of the transcribed spacers in the nascent pre-rRNA. In this study the RAC-binding site in the internal transcribed spacer 2 sequence of Schizosaccharomyces pombe rRNA transcripts was identified, and the influence of this site on rRNA maturation was assessed. Modification exclusion analyses indicate that the protein complex interacts with a helical domain previously shown to contain features common to both the internal transcribed spacer 1 and the 3'-external transcribed spacer. Mutagenic analyses in vitro confirm an interaction with this sequence, and parallel analyses in vivo indicated a critical role in both the maturation of the rRNA components of the large subunit as well as the 18 S rRNA component of the small subunit. Hybridization analyses also indicated greatly elevated levels of unprocessed nascent RNA. These effects are contrasted with mutations in other regions of the secondary structure that resulted in some reduction of plasmid-derived mature rRNA but no elevated levels of the precursor molecules. The significance with respect to rRNA maturation and the interdependences in rRNA processing are discussed. PMID- 11923315 TI - Repair of clustered DNA lesions. Sequence-specific inhibition of long-patch base excision repair be 8-oxoguanine. AB - Ionizing radiation induces clustered DNA damage where two or more lesions are located proximal to each other on the same or opposite DNA strands. It has been suggested that individual lesions within a cluster are removed sequentially and that the presence of a vicinal lesion(s) may affect the rate and fidelity of DNA repair. In this study, we addressed the question of how 8-oxoguanine located opposite to normal or reduced abasic sites would affect the repair of these sites by the base excision repair system. We have found that an 8-oxoguanine located opposite to an abasic site does not affect either the efficiency or fidelity of repair synthesis by DNA polymerase beta. In contrast, an 8-oxoguanine located one nucleotide 3'-downstream of the abasic site significantly reduces both strand displacement synthesis supported by DNA polymerase beta or delta and cleavage by flap endonuclease of the generated flap, thus inhibiting the long-patch base excision repair pathway. PMID- 11923316 TI - hsp90 is required for heme binding and activation of apo-neuronal nitric-oxide synthase: geldanamycin-mediated oxidant generation is unrelated to any action of hsp90. AB - It is established that neuronal NO synthase (nNOS) is associated with the chaperone hsp90, although the functional role for this interaction has not been defined. We have discovered that inhibition of hsp90 by radicicol or geldanamycin nearly prevents the heme-mediated activation and assembly of heme-deficient apo nNOS in insect cells. This effect is concentration-dependent with over 75% inhibition achieved at 20 microm radicicol. The ferrous carbonyl complex of nNOS is not formed when hsp90 is inhibited, indicating that functional heme insertion is prevented. We propose that the hsp90-based chaperone machinery facilitates functional heme entry into apo-nNOS by the opening of the hydrophobic heme binding cleft in the protein. Previously, it has been reported that the hsp90 inhibitor geldanamycin uncouples endothelial NOS activity and increases endothelial NOS-dependent O(2)() production. Geldanamycin is an ansamycin benzoquinone, and we show here that it causes oxidant production from nNOS in insect cells as well as with the purified protein. At a concentration of 20 microm, geldanamycin causes a 3-fold increase in NADPH oxidation and hydrogen peroxide formation from purified nNOS, whereas the non-quinone hsp90 inhibitor radicicol had no effect. Thus, consistent with the known propensity of other quinones, geldanamycin directly redox cycles with nNOS by a process independent of any action on hsp90, cautioning against the use of geldanamycin as a specific inhibitor of hsp90 in redox-active systems. PMID- 11923317 TI - Plasticity of recognition of the 3'-end of mischarged tRNA by class I aminoacyl tRNA synthetases. AB - Certain aminoacyl-tRNA synthetases prevent potential errors in protein synthesis through deacylation of mischarged tRNAs. For example, the close homologs isoleucyl-tRNA synthetase (IleRS) and valyl-tRNA synthetase (ValRS) deacylate Val tRNA(Ile) and Thr-tRNA(Val), respectively. Here we examined the chemical requirements at the 3'-end of the tRNA for these hydrolysis reactions. Single atom substitutions at the 2'- and 3'-hydroxyls of a variety of mischarged RNAs revealed that, while acylation is at the 2'-OH for both enzymes, IleRS catalyzes deacylation specifically from the 3'-OH and not from the 2'-OH. In contrast, ValRS can deacylate non-cognate amino acids from the 2'-OH. Moreover, for IleRS the specificity for a 3'-O location of the scissile ester bond could be forced to the 2'-position by introduction of a 3'-O-methyl moiety. Cumulatively, these and other results suggest that the editing sites of these class I aminoacyl-tRNA synthetases have a degree of inherent plasticity for substrate recognition. The ability to adapt to subtle differences in mischarged RNAs may be important for the high accuracy of aminoacylation. PMID- 11923318 TI - Cancer-associated cleavage of cytokeratin 8/18 heterotypic complexes exposes a neoepitope in human adenocarcinomas. AB - The intermediate filament network in simple glandular epithelial cells predominantly consists of heterotypic complexes of cytokeratin 8 (K8) and cytokeratin 18 (K18). In contrast to other cytokeratins, K8 and K18 are persistently expressed during malignant transformation, but changes in cell morphology are accompanied by alterations in the intermediate filament network. To study molecular changes, K8 and K18 were purified from surgically removed colon cancer and normal epithelia tissues. Western blotting and amino acid sequencing revealed the presence of abundant K8 and K18 fragments, truncated at the N terminus, from cancerous, but not normal, epithelial cells. The fragmentation pattern indicates proteolysis mediated by several enzymes, including trypsin-like enzymes. The cancer-associated forms of K8 and K18 are specifically recognized by the human antibody, COU-1, cloned from the B cells of a cancer patient. We demonstrate that COU-1 recognizes a unique conformational epitope presented only by a complex between K8 and K18. The epitope is revealed after proteolytic removal of the head domain of either K8 or K18. A large panel of recombinant K8 and K18 fragments, deleted N- or C-terminally, allowed for the localization of the COU-1 epitope to the N-terminal part of the rod domains. Using surface plasmon resonance, the affinity of COU-1 for this epitope was determined to be 10(9) x m(-1), i.e. more than 2 orders of magnitude higher than for intact heterotypic K8/K18 complexes. The cellular distribution of truncated K8/K18 heterotypic complexes in viable adenocarcinomas cells was probed using COU 1 showing small fibrillar structures distinct from those of intact K8/K18 complexes. Previously we demonstrated the binding and subsequent internalization of recombinant Fab COU-1 to live cancer cells. We have thus characterized a cancer neoepitope recognized by the humoral immune system. The results have biological as well as clinical implications. PMID- 11923320 TI - Miscellaneous catalase-negative, gram-positive cocci: emerging opportunists. PMID- 11923319 TI - Regulation of the Saccharomyces cerevisiae Slt2 kinase pathway by the stress inducible Sdp1 dual specificity phosphatase. AB - The Slt2/Mpk1 mitogen-activated protein kinase (MAPK) cell integrity pathway is involved in maintenance of cell shape and integrity during vegetative growth and mating in Saccharomyces cerevisiae. Slt2 is activated by dual phosphorylation of a threonine and tyrosine residue in response to several environmental stresses that perturb cell integrity. Negative regulation of Slt2 is achieved via dephosphorylation by two protein-tyrosine phosphatases, Ptp2 and Ptp3, and a dual specificity phosphatase, Msg5. In this study, we provide genetic and biochemical evidence that the stress-inducible dual specificity phosphatase, Sdp1, negatively regulates Slt2 by direct dephosphorylation. Deletion of SDP1 exacerbated growth defects due to overexpression of Mkk1(p386), a constitutively active mutant of Slt2 MAPK kinase, whereas overexpression of Sdp1 suppressed lethality caused by Mkk1(p386) overexpression. The heat shock-induced phosphorylation level of Slt2 was elevated in an sdp1Delta strain compared with that of the wild type, and heat shock-activated phospho-Slt2 was dephosphorylated by recombinant Sdp1 in vitro. Under normal growth conditions, an Sdp1-GFP fusion protein was localized to both the nucleus and cytoplasm. However, the Sdp1-GFP protein translocated to punctate spots throughout the cell after heat shock. SDP1 transcription was induced by several stress conditions in an Msn2/4-dependent manner but independent of the Rlm1 transcription factor, a downstream target activated by Slt2. Induction of SLT2 by high osmolarity was dependent on Rlm1 transcription factor and Hog1 kinase, suggesting cross-talk between Slt2 and Hog1 MAPK pathways. These studies demonstrate regulation of Slt2 activity and gene expression in coordination with other stress signaling pathways. PMID- 11923321 TI - Pseudoepidemic due to a unique strain of Mycobacterium szulgai: genotypic, phenotypic, and epidemiological analysis. AB - In mid-1999, we noted multiple isolations at the Veterans Affairs Medical Center (VAMC) Houston Tex. of an unusual nonpigmented Mycobacterium species. Since, on the basis of 16S rRNA gene sequence analysis, the strains were identical to the Mycobacterium szulgai type strain and since M. szulgai has been reported only rarely as a commensal or environmental isolate, we were concerned about laboratory contamination, nosocomial spread, or even the possibility that this could be a novel organism associated with disease. Our investigation found that from 1999 to 2000, 37 strains of M. szulgai were isolated from patients at the VAMC (the base rate for the previous 10 years had been <1 isolation per year). We compared the phenotypic properties and genetic relatedness of these 37 strains (31 of which were nonpigmented) as well as eight stock strains and the M. szulgai type strain. All strains were similar in cellular fatty acid patterns, growth rates, and biochemical characteristics. However, we found three genogroups by gene sequence analysis. Genogroup I comprised the M. szulgai type strain, all the tested nonpigmented strains (27 of the 31 strains were tested), two pigmented strains isolated in 1999 and 2000, and five pigmented stock strains. Genogroup II comprised five pigmented strains: three were isolated from 1999 to 2000 and two were stock strains. The single strain (isolated in 1996) in genogroup III was pigmented and was the only strain associated with disease. Whereas the randomly amplified polymorphic DNA (RAPD) patterns of all nonpigmented strains were identical, indicating that they came from a common source (the pseudoepidemic strain), the RAPD patterns of the other strains were varied. In our investigation for a possible source, we found that there were no common reagents, specimen processing or patient locations, or procedures linking the 31 pseudoepidemic strains. However, a nonpigmented M. szulgai strain with a gene sequence and RAPD pattern identical to those of the pseudoepidemic strain was recovered from a water storage tank serving the hospital. We concluded that the strains most likely originated from hospital water, which transiently inoculated our patients. Although no disease was associated with this cluster of isolates, the event was costly because identification was problematic and we could not easily discount the isolations, since most of the patients were immunocompromised and were candidates for opportunistic infection. PMID- 11923322 TI - Enterococcus gilvus sp. nov. and Enterococcus pallens sp. nov. isolated from human clinical specimens. AB - Light yellow-pigmented (strain PQ1) and yellow-pigmented (strain PQ2), gram positive, non-spore-forming, nonmotile bacteria consisting of pairs or chains of cocci were isolated from the bile of a patient with cholecystitis (PQ1) and the peritoneal dialysate of another patient with peritonitis (PQ2). Morphologically and biochemically, the organisms phenotypically belonged to the genus Eterococcus. Whole-cell protein (WCP) analysis and sequence analysis of a segment of the 16S rRNA gene suggested that they are new species within the genus Enterococcus. PQ1 and PQ2 displayed less than 70% identities to other enterococcal species by WCP analysis. Sequence analysis showed that PQ1 shared the highest level of sequence similarity with Enterococcus raffinosus and E. malodoratus (sequence similarities of 99.8% to these two species). Sequence analysis of PQ2 showed that it had the highest degrees of sequence identity with the group I enterococci E. malodoratus (98.7%), E. raffinosus (98.6%), E. avium (98.6%), and E. pseudoavium (98.6%). PQ1 and PQ2 can be differentiated from the other Enterococcus spp. in groups II, III, IV, and V by their phenotypic characteristics: PQ1 and PQ2 produce acid from mannitol and sorbose and do not hydrolyze arginine, placing them in group I. The yellow pigmentation differentiates these strains from the other group I enterococci. PQ1 and PQ2 can be differentiated from each other since PQ1 does not produce acid from arabinose, whereas PQ2 does. Also, PQ1 is Enterococcus Accuprobe assay positive and pyrrolidonyl-beta-naphthylamide hydrolysis positive, whereas PQ2 is negative by these assays. The name Enterococcus gilvus sp. nov. is proposed for strain PQ1, and the name Enterococcus pallens sp. nov. is proposed for strain PQ2. Type strains have been deposited in culture collections as E. gilvus ATCC BAA-350 (CCUG 45553) and E. pallens ATCC BAA-351 (CCUG 45554). PMID- 11923323 TI - Genetic diversity of Pneumocystis carinii f. sp. hominis based on variations in nucleotide sequences of internal transcribed spacers of rRNA genes. AB - A variety of genes have been used to type Pneumocystis carinii. In the present study, nucleotide sequence variations in the ITS1 and ITS2 internal transcribed spacer (ITS) regions of the rRNA genes were used to type Pneumocystis carinii f. sp. hominis DNA obtained from the lungs of 60 human immunodeficiency virus infected individuals. These regions were amplified by PCR, cloned, and sequenced. Multibase polymorphisms were identified among samples. Several new genotypes are reported on the basis of the nucleotide sequence variations at previously unreported positions of both the ITS1 and the ITS2 regions. Twelve new ITS1 sequences were observed, in addition to the nine sequence types reported previously. The most common was type E, which was observed in 60.5% of the samples. The sequence variations in the ITS1 region were mainly located at positions 5, 12, 23, 24, 45, 53, and 54. Sixteen new ITS2 types were also identified, in addition to the 13 types reported previously. The most common was type g (26.6%). The sequences of the ITS2 regions in most specimens were different from the previously published sequence at bases 120 and 166 through 183. The most common variations observed were deletions at positions 177 through 183. The presence of more than one sequence type in some patients (60%) suggested the occurrence of coinfection with multiple P. carinii strains. The genetic polymorphism observed demonstrates the degree of diversity of Pneumocystis strains that infect humans. Furthermore, the high degree of polymorphism suggests that these genes are evolving faster than other genes. Consequently, the sequence information derived is useful for purposes such as examination of the potential of person-to-person transmission and recurrent infections but perhaps not for other genotyping applications that rely on more stable genetic loci. PMID- 11923324 TI - Polymorphic amplified typing sequences provide a novel approach to Escherichia coli O157:H7 strain typing. AB - Escherichia coli O157:H7 (O157) strains are commonly typed by pulsed-field gel electrophoresis (PFGE) following digestion of genomic DNA with the restriction enzyme XbaI. We have shown that O157 strains differ from each other by a series of discrete insertions or deletions, some of which contain recognition sites for XbaI, suggesting that these insertions and deletions are responsible for the differences in PFGE patterns. We have devised a new O157 strain typing protocol, polymorphic amplified typing sequences (PATS), based on this information. We designed PCR primer pairs to amplify genomic DNA flanking each of 40 individual XbaI sites in the genomes of two O157 reference strains. These primer pairs were tested with 44 O157 isolates, 2 each from 22 different outbreaks of infection. Thirty-two primer pairs amplified identical fragments from all 44 isolates, while eight primer pairs amplified regions that were polymorphic between isolates. The isolates could be differentiated solely on the basis of which of the eight polymorphic amplicons was detected. PATS correctly identified 21 of 22 outbreak pairs as being identical or highly related, whereas PFGE correctly identified 14 of the 22 outbreak pairs as being identical or highly related; PATS was also able to type isolates from three outbreaks that were untypeable by PFGE. However, PATS was less sensitive than PFGE in discriminating between outbreaks. These data suggest that typing by PATS may provide a simple procedure for strain typing of O157 and other bacteria and that further evaluation of the utility of this method for epidemiologic investigations is warranted. PMID- 11923325 TI - Molecular characterization of vancomycin-resistant Enterococci repopulating the gastrointestinal tract following treatment with a novel glycolipodepsipeptide, ramoplanin. AB - We characterized baseline and repopulating stool isolates recovered during a phase II trial of ramoplanin for the treatment of patients with stool carriage of vancomycin-resistant enterococci (VRE). Repopulation with a strain with a related genotype was found in 74, 60, and 53% of individuals in groups treated with placebo, 100 mg of ramoplanin, and 400 mg of ramoplanin, respectively. All ramoplanin-treated patients with a culture positive for VRE at day 7 had a relapse caused by a genotypically related isolate. In ramoplanin-treated patients, antibiotics with activities against anaerobic organisms were associated with positive cultures on day 7 (relative risk [RR] = 8.8; P = 0.004), and the avoidance of such antibiotics was significantly associated with culture negativity through day 21 (RR = 0.16; P = 0.02). PMID- 11923326 TI - Genotyping of a homogeneous group of Yersinia pestis strains isolated in the United States. AB - Yersinia pestis, the causative agent of deadly plague, is considered a reemerging infectious disease and a significant biological terrorism threat. The present project focused on epidemiological investigation of the genetic variability of well-documented strains of Y. pestis from the United States by pulsed-field gel electrophoresis (PFGE) and restriction fragment length polymorphism (RFLP) analysis with insertion sequences IS100 and IS285 as probes. We examined 37 U.S. Y. pestis strains and isolates of a single ribotype, ribotype B, recovered between 1939 and 1998 from patients, animals, and fleas. Our results showed that all isolates had similar PFGE patterns, but minor differences such as missing, additional, and shifted bands were found among almost all strains if they came from different parent strains. The 37 strains and isolates were divided into 26 PFGE types. RFLP analysis with IS100 as a probe divided these strains and isolates into 16 types, with 43% belonging to IS100 type 1. Typing with IS285 as a probe was less specific and led to only four RFLP types, with 81% belonging to type 1. Similarity analysis with BioNumerics software showed that all strains shared >or=80, 86, and 91% similarities on dendrograms prepared from digitized PFGE, IS100 RFLP analysis, and IS285 RFLP analysis images, respectively. Our results demonstrate that PFGE offers an increased ability to discriminate between strains (Simpson's index of diversity, 0.98) and therefore can significantly improve epidemiological studies related to the origin of new plague isolates. PMID- 11923327 TI - Recombinant BBK32 protein in serodiagnosis of early and late Lyme borreliosis. AB - Borrelial protein BBK32 was evaluated as an antigen in the serodiagnosis of early and disseminated Lyme borreliosis (LB). bbk32 was cloned and sequenced from eight isolates of the three pathogenic Borrelia species. The identities between the amino acid sequences of the BBK32 proteins from Borrelia burgdorferi sensu stricto, B. garinii, and B. afzelii isolates were 71 to 100%. By immunoglobulin G (IgG) Western blotting (WB) or enzyme-linked immunosorbent assay (ELISA), up to 74 and 100% of acute- and convalescent-phase samples, respectively, from 23 patients with erythema migrans (EM) were positive for recombinant BBK32 protein from B. afzelii. In the serology of disseminated LB, the three variant BBK32 antigens cross-reacted. In total, 14 of 14 samples from patients with neuroborreliosis and 15 of 15 samples from patients with Lyme arthritis were positive. The specificities of the IgG ELISA with the variant BBK32 antigens for EM and disseminated borreliosis were 81 to 92% and 89 to 95%, respectively. Our findings indicate that the BBK32 proteins are promising serodiagnostic antigens for the detection of early and disseminated LB but that variant BBK32 proteins may be needed either in parallel or in combination with an immunoassay for LB to cover all the relevant borrelial species that cause the disease. PMID- 11923329 TI - Six-year molecular analysis of Burkholderia cepacia complex isolates among cystic fibrosis patients at a referral center for lung transplantation. AB - Over a 6-year period, Burkholderia cepacia complex species were isolated from cystic fibrosis (CF) patients receiving care at The University of North Carolina Hospitals (clinic CF patients) and from those referred from other treatment centers. Fifty-six isolates collected from 30 referred patients and 26 clinic CF patients were characterized by pulsed-field gel electrophoresis (PFGE) and were assayed by PCR to detect the cable pilin gene, cblA. PFGE results indicated that six separate clusters (clusters A to F) were present among the 56 isolates and that three clusters (clusters A, B, and E) consisted only of isolates from referred patients infected with B. cepacia complex isolates prior to referral. However, one cluster (cluster C) consisted of isolates from four CF patients, and hospital records indicate that this cluster began with an isolate that came from a referred patient and that spread to three clinic CF patients. Cluster D consisted of two isolates from clinic CF patients, and hospitalization records are consistent with nosocomial, patient-to-patient spread. cblA was present in only 4 of the 56 isolates and included isolates in cluster E from the referred patients. Our results indicate a lack of spread of a previously characterized, transmissible clone from referred patients to our clinic CF population. Only two instances of nosocomial, patient-to-patient spread could be documented over the 6 year period. An additional spread of an isolate (cluster F) from a referred patient to a clinic patient could not be documented as nosocomial and may have been the result of spread in a nonhospitalized setting. The majority (36 of 56) of our B. cepacia complex-infected CF patients harbor isolates with unique genotypes, indicating that a diversity of sources account for infection. These data suggest that CF patients infected with B. cepacia complex and referred for lung transplantation evaluation were not a major source of B. cepacia complex strains that infected our resident CF clinic population. PMID- 11923328 TI - Population structure and genetic diversity of Actinobacillus actinomycetemcomitans strains isolated from localized juvenile periodontitis patients. AB - The phylogeny of 20 Actinobacillus actinomycetemcomitans strains isolated from patients with localized juvenile periodontitis (LJP) was investigated by using partial sequence analysis of 16S rRNA genes, arbitrarily primed PCR (AP-PCR), and four additional PCR assays that amplified polymorphic regions in the leukotoxin (lkt), cytolethal distending toxin (cdt), major fimbrial subunit (flp-1), and serotype-specific O polysaccharide gene clusters. Our analysis also included four strains isolated from healthy subjects and nine reference strains. We found that A. actinomycetemcomitans strains comprised three major phylogenetic lineages. One lineage consisted of serotype b strains, a second lineage consisted of serotype c strains, and a third lineage consisted of serotype a, d, e, and f strains. 16S rRNA sequences within each lineage were highly conserved (<1% base substitutions), whereas sequences between lineages were exceptionally divergent (1.9 to 5.0% substitutions). Two strains exhibited 16S rRNA sequences that were even more distantly related to those of the three major lineages (2.7 to 6.7% substitutions), indicating that additional minor lineages or variants exist. The distribution of 16S rRNA sequences and lkt, cdt, flp-1, and AP-PCR genotypes was consistent with a clonal population structure, with little evidence of assortative recombination between strains of different serotypes. Strains from all three major lineages were recovered from LJP patients, suggesting that phylogenetically diverse strains of A. actinomycetemcomitans carry pathogenic potential. PMID- 11923330 TI - Quantitative detection of Neospora caninum in bovine aborted fetuses and experimentally infected mice by real-time PCR. AB - We report the development of a real-time PCR assay for the quantitative detection of Neospora caninum in infected host tissues. The assay uses the double-stranded DNA-binding dye SYBR Green I to continuously monitor product formation. Oligonucleotide primers were designed to amplify a 76-bp DNA fragment corresponding to the Nc5 sequence of N. caninum. A similar method was developed to quantify the 28S rRNA host gene in order to compare the parasite load of different samples and to correct for the presence of potential PCR-inhibiting compounds in the DNA samples. A linear quantitative detection range of 6 logs with a calculated detection limit of 10(-1) tachyzoite per assay was observed with excellent linearity (R(2) = 0.998). Assay specificity was confirmed by using DNA from the closely related parasite Toxoplasma gondii. The applicability of the technique was successfully tested in a variety of host brain tissues: (i) aborted bovine fetuses classified into negative or positive Neospora-infected animals according to the observation of compatible lesions by histopathological study and (ii) experimentally infected BALB/c mice, divided into three groups, inoculated animals with or without compatible lesions and negative controls. All samples were also tested by ITS1 Neospora nested PCR and a high degree of agreement was shown between both PCR techniques (kappa = 0.86). This technique represents a useful quantitative diagnostic tool to be used in the study of the pathogenicity, immunoprophylaxis, and treatment of Neospora infection. PMID- 11923331 TI - Mitochondrial DNA restriction fragment length polymorphism (RFLP) and 18S small subunit ribosomal DNA PCR-RFLP analyses of Acanthamoeba isolated from contact lens storage cases of residents in southwestern Korea. AB - We applied ribosomal DNA PCR-restriction fragment length polymorphism (RFLP) and mitochondrial DNA (mtDNA) RFLP analyses to 43 Acanthamoeba environmental isolates (KA/LH1 to KA/LH43) from contact lens storage cases in southwestern Korea. These isolates were compared to American Type Culture Collection strains and clinical isolates (KA/E1 to KA/E12) from patients with keratitis. Seven riboprint patterns were seen. To identify the species of the isolates, a phylogenetic tree was constructed based on the comparison of riboprint patterns with reference strains. Four types accounted for 39 of the isolates belonging to the A. castellanii complex. The most predominant (48.8%) type was A. castellanii KA/LH2 type, which had identical riboprint and mtDNA RFLP patterns to those of A. castellanii Castellani, KA/E3 and KA/E8. The riboprint and mtDNA RFLP patterns of the KA/LH7 (20.9%) type were identical to those of A. castellanii Ma, a corneal isolate from the United States. The riboprint and mtDNA RFLP patterns of the KA/LH1 (18.6%) type were the same as those of A. lugdunensis L3a, KA/E2, and KA/E12. The prevalent pattern for each type of Acanthamoeba in southwestern Korea was very different from that from southeastern Korea and Seoul, Korea. It is noteworthy that 38 (88.4%) out of 43 isolates from contact lens storage cases of the residents in southwestern Korea revealed mtDNA RFLP and riboprint patterns identical to those found for clinical isolates in our area. This indicates that most isolates from contact lens storage cases in the surveyed area are potential keratopathogens. More attention should be paid to the disinfection of contact lens storage cases to prevent possible amoebic keratitis. PMID- 11923332 TI - Genotypes and clinical phenotypes of hepatitis B virus in patients with chronic hepatitis B virus infection. AB - Genotype C of hepatitis B virus (HBV) has been shown to be associated with a poor clinical outcome, compared to genotype B. To explore the clinical phenotypes, with special reference to the seroconversion of hepatitis B e antigen (HBeAg) and frequency of acute exacerbation between patients infected with HBV genotypes B and C, a cohort of 272 Taiwanese patients with chronic HBV infection was analyzed. According to the status of HBeAg at enrollment and frequency of acute exacerbation during the follow-up period, five groups of patients with distinct clinical phenotypes were categorized. Of the 272 HBV carriers, 185 (68%) were infected with HBV genotype B and the remaining 87 (32%) were infected with genotype C. Among them, 150 (55%) were positive for HBeAg and patients with genotype C infection tended to have a higher positive rate of HBeAg than those with genotype B infection (63 versus 51%). Genotype B was more prevalent than genotype C in different groups of HBV carriers. However, the prevalence of genotype C in patients with multiple episodes of acute exacerbation who failed to have HBeAg seroconversion was significantly higher than in all 272 patients (50 versus 32%, P = 0.025), in those with HBeAg seroconversion after only one episode of acute exacerbation (50 versus 12%, P = 0.01), or in those negative for HBeAg at enrollment and without acute exacerbations (50 versus 23%, P = 0.002). In conclusion, patients with genotype C infection have a more aggressive clinical phenotype than do those with genotype B infection, which contributes to the former group's progressive liver disease and poor clinical outcomes. PMID- 11923333 TI - Ribosomal DNA-directed PCR for identification of Achromobacter (Alcaligenes) xylosoxidans recovered from sputum samples from cystic fibrosis patients. AB - The opportunistic human pathogen Achromobacter (Alcaligenes) xylosoxidans has been recovered with increasing frequency from respiratory tract culture of persons with cystic fibrosis (CF). However, confusion of this species with other closely related respiratory pathogens has limited studies to better elucidate its epidemiology, natural history, and pathogenic role in CF. Misidentification of A. xylosoxidans as Burkholderia cepacia complex is especially problematic and presents a challenge to effective infection control in CF. To address the problem of accurate identification of A. xylosoxidans, we developed a PCR assay based on a 16S ribosomal DNA sequence. In an analysis of 149 isolates that included 47 A. xylosoxidans and several related glucose-nonfermenting species recovered from CF sputum, the sensitivity and specificity of this PCR assay were determined to be 100 and 97%, respectively. The availability of this assay will enhance identification of A. xylosoxidans, thereby facilitating study of the pathogenic role of this species and improving infection control efforts in CF. PMID- 11923335 TI - Relatively alcohol-resistant mycobacteria are emerging pathogens in patients receiving acupuncture treatment. AB - Acupuncture has been gaining popularity as a form of alternative medicine. In the past, only blood-borne viruses and anecdotal reports of bacterial infections have been associated with acupuncture. We report on four patients with mycobacterial infections complicating acupuncture who were encountered in a 2-year period. All had clinical and/or radiological lesions at acupuncture point- and meridian specific locations. There was no other history of trauma or other clinical foci of infections, and the chest radiographs were normal. Histological studies of biopsy specimens of all four patients showed changes compatible with chronic inflammation, with granulomatous inflammation present in three patients and acid fast bacilli present in two. Conventional biochemical tests and whole-cell fatty acid analysis for identification were inconclusive for all four nonpigmented mycobacteria recovered from tissue biopsies. 16S rRNA gene sequencing showed that the strains from two patients were Mycobacterium chelonae and that those from the other two were Mycobacterium nonchromogenicum. Alcohol resistance assay using the quantitative suspension test revealed that all four strains showed prolonged survival in 75% alcohol compared to other skin flora. Mycobacterial infections transmitted by acupuncture are an emerging problem. A high index of suspicion is essential to recognize this clinical syndrome, and strict implementation of proper infection control guidelines for acupuncture is mandatory. PMID- 11923334 TI - Laccase and melanization in clinically important Cryptococcus species other than Cryptococcus neoformans. AB - The laccase enzyme and melanin synthesis have been implicated as contributors to virulence in Cryptococcus neoformans. Since isolations of Cryptococcus species other than C. neoformans from clinical specimens have been increasing, we examined the laccase activities of C. albidus, C. laurentii, C. curvatus, and C. humicola. Incubation of cells with epinephrine produced adrenochrome color in C. albidus, C. laurentii, and C. curvatus but not in C. humicola. Activity was always less than in C. neoformans. Laccase was detected in the soluble fractions of disrupted C. albidus, C. laurentii, and C. curvatus cells. Activity staining of partially purified enzyme after nondenaturing polyacrylamide gel electrophoresis revealed that laccases from C. albidus, C. laurentii, and C. curvatus migrated more slowly than that from C. neoformans. One strain of C. curvatus exhibited two melanin bands. Thus, several clinically emerging Cryptococcus species express laccase and can synthesize melanin. PMID- 11923336 TI - Escherichia coli ehl1 gene-positive serotype O18ac:H31 associated with an outbreak of diarrhea in a neonatal nursery in Neuquen City, Argentina. AB - Between 9 October and 12 November 1996, an outbreak of bloody diarrhea occurred in the neonatal nursery ward of the Policlinico Neuquen, in Neuquen, a city in the southwestern region of Argentina. Seven patients of the intermediate care unit were affected. Isolates of Escherichia coli O18ac:H31 that were non-lactose and -sorbitol fermenting were recovered from outbreak cases. Although the strains were negative for a number of virulence factors typically found in diarrheagenic groups of E. coli, all isolates from the present neonatal outbreak possessed the enterohemolysin gene, ehl1. All isolates showed resistance to the antibiotics ampicillin and chloramphenicol. These isolates showed a low adherence property in HeLa cells without any recognizable pattern. In order to evaluate the outbreak dissemination in the neonatology ward, a prevalence study was conducted on 13 November. Stool specimens were obtained from 16 neonates hospitalized in the sector and from 33 medical staff members. E. coli isolates with identical biochemical characteristics of the outbreak strain were recovered from 11 of 16 inpatients and from 4 of 33 staff members during the prevalence study. A total of 15 E. coli strains recovered both from the outbreak and the prevalence study were processed by random amplified polymorphic DNA (RAPD)-PCR and pulsed-field gel electrophoresis (PFGE). By RAPD-PCR 14 of 15 strains showed patterns with 85 to 100% similarity, and by PFGE these strains were identical, each showing a unique pattern with 15 bands between 40 and 400 kb. One strain isolated from a nurse during the prevalence study presented a pattern not related to the others, and this was characterized as E. coli O81:HNM resistant to ampicillin only and negative for all the virulence factors studied. This outbreak occurred despite strict regulations in place to prevent cross-infection in the hospital. Postoutbreak prevalence studies were performed weekly thereafter without detecting new cases. PMID- 11923337 TI - Phenotypic and molecular characterization of clinical isolates of Mycobacterium elephantis from human specimens. AB - Eleven strains of a rapidly growing mycobacterium were isolated from patient specimens originating from various regions of the province of Ontario, Canada, over a 2-year period. Unique high-performance liquid chromatography (HPLC) and PCR-restriction enzyme pattern analysis (PRA) profiles initially suggested a new Mycobacterium species, while sequencing of the 16S rRNA gene revealed a sequence match with Mycobacterium sp. strain MCRO 17 (GenBank accession no. X93028), an isolate determined to be unique which is to date uncharacterized, and also a close similarity to M. elephantis (GenBank accession no. AJ010747), with six base pair variations. A complete biochemical profile of these isolates revealed a species of mycobacteria with phenotypic characteristics similar to those of M. flavescens. HPLC, PRA, and 16S rRNA sequencing of strain M. elephantis DSM 44368(T) and result comparisons with the clinical isolates revealed that these strains were in fact M. elephantis, a newly described species isolated from an elephant. All strains were isolated from human samples, 10 from sputum and 1 from an axillary lymph node. PMID- 11923339 TI - Biofilm production by isolates of Candida species recovered from nonneutropenic patients: comparison of bloodstream isolates with isolates from other sources. AB - Biofilm production has been implicated as a potential virulence factor of some Candida species responsible for catheter-related fungemia in patients receiving parenteral nutrition. We therefore compared clinical bloodstream isolates representing seven different Candida species to each other and to those from other anatomical sites for the capacity to form biofilms in glucose-containing medium. Potential associations between the capacity to form biofilms and the clinical characteristics of fungemia were also analyzed. Isolates included the following from nonneutropenic patients: 101 bloodstream isolates (35 C. parapsilosis, 30 C. albicans, 18 C. tropicalis, 8 C. glabrata, and 10 other Candida species isolates) and 259 clinical isolates from other body sites (116 C. albicans, 53 C. glabrata, 43 C. tropicalis, 17 C. parapsilosis, and 30 other Candida species isolates). Organisms were grown in Sabouraud dextrose broth (SDB) containing a final concentration of 8% glucose to induce biofilm formation, as published previously. Biofilm production was determined by both visual and spectrophotometric methods. In this medium, biofilm production by C. albicans isolates was significantly less frequent (8%) than that by non-C. albicans Candida species (61%; P < 0.0001). The overall proportion of non-C. albicans Candida species isolates from the blood that produced biofilms was significantly higher than that of non-C. albicans Candida isolates obtained from other sites (79% versus 52%; P = 0.0001). Bloodstream isolates of C. parapsilosis alone were significantly more likely to be biofilm positive than were C. parapsilosis isolates from other sites (86% versus 47%; P = 0.0032). Non-C. albicans Candida species, including C. parapsilosis, were more likely to be biofilm positive if isolates were derived from patients whose candidemia was central venous catheter (CVC) related (95%; P < 0.0001) and was associated with the use of total parenteral nutrition (TPN) (94%; P < 0.005). These data suggest that the capacity of Candida species isolates to produce biofilms in vitro in glucose-containing SDB may be a reflection of the pathogenic potential of these isolates to cause CVC-related fungemia in patients receiving TPN. PMID- 11923338 TI - Epidemiology of extended-spectrum beta-lactamase-producing Enterobacter isolates in a Spanish hospital during a 12-year period. AB - Fifteen Enterobacter clinical isolates (11 Enterobacter cloacae isolates, 3 Enterobacter aerogenes isolates, and 1 Enterobacter gergoviae isolate), representing 0.4% of all Enterobacter isolates recovered in our hospital from 1989 to 2000, were suspected of harboring an extended-spectrum beta-lactamase (ESBL). These isolates were recovered from 14 different patients. ESBLs were transferred by conjugation into an Escherichia coli recipient strain. Pulsed field gel electrophoresis (PFGE) revealed a single clone of E. aerogenes and six different clones of E. cloacae. Four of these E. cloacae clonal types were represented by only one isolate each, but the other two were represented by three and four isolates, respectively. Isoelectric focusing, susceptibility phenotyping, PCR analysis, and sequencing demonstrated the presence of three different ESBLs. The most frequent was the recently characterized CTX-M-10 ESBL, which was found in the E. gergoviae isolate and in all but one of the E. cloacae isolates. The remaining E. cloacae isolate harbored a TEM-27 ESBL, and the three E. aerogenes isolates harbored a TEM-24 ESBL. PFGE revealed that our E. aerogenes strain was indistinguishable from the French TEM-24-producing E. aerogenes endemic clone. Although a low prevalence of ESBL-producing Enterobacter isolates was found in our institution over a 12-year period, a diversity of nonepidemic E. cloacae clones was detected, as was the persistence of the CTX-M-10 beta lactamase. The presence of the TEM-24-producing E. aerogenes French clone in our institution also demonstrates the intercountry dissemination of ESBL-producing isolates. PMID- 11923340 TI - Quantitative detection of Borrelia burgdorferi in 2-millimeter skin samples of erythema migrans lesions: correlation of results with clinical and laboratory findings. AB - Variability of disease manifestations has been noted in patients with Lyme disease. A contributing factor to this variation may be the number of spirochetes present in infected patients. We evaluated clinical and laboratory findings for patients with erythema migrans with regard to the number of Borrelia burgdorferi organisms detected by quantitative PCR (qPCR) in 2-mm skin biopsy specimens. B. burgdorferi was detected in 80% (40 of 50) of the specimens tested; the mean number of spirochetes in these specimens ranged over 3 orders of magnitude (10 to 11,000 spirochetes per 2-mm biopsy specimen). Larger numbers of spirochetes were significantly associated with a shorter duration of the erythema migrans skin lesion (P = 0.020), smaller skin lesions (P = 0.020), and infection with a specific genotype of B. burgdorferi (P = 0.008) but not with the number or severity of symptoms. Skin culture positivity was significantly associated with skin lesions containing larger numbers of spirochetes (P = 0.019). PMID- 11923341 TI - Comparison of DNA hybridization and PCR assays for detection of putative pathogenic enteroadherent Escherichia coli. AB - The correlation of the different adherence patterns with DNA probes and PCR primers for the identification of Escherichia coli was analyzed in isolates from children, less than 2 years of age with or without diarrhea, from different regions of Brazil. A total of 1,428 isolates obtained from 338 patients and 322 control children were studied. The enteropathogenic E. coli (EPEC) adherence factor (EAF) probe was shown to be as good as the HEp-2 adhesion assay for the detection of typical EPEC strains. The DNA probes used to detect diffusely adhering E. coli and enteroaggregative E. coli (EAEC) showed low sensitivities (64 and 50%, respectively), and the best method of identifying these organisms in clinical research remains the HEp-2 adherence assay. The "bundle-forming pilus" (BFP) and the EAEC PCR assays could be used instead of the DNA probes as a screening method for typical EPEC and EAEC carrying the EAEC probe sequence in the clinical laboratory. In our study, only typical EPEC strains that carried EAF and BFP were associated with acute diarrhea. PMID- 11923342 TI - Transmission studies of Babesia microti in Ixodes ricinus ticks and gerbils. AB - In order to investigate the possible role of Ixodes ricinus as a vector of zoonotic Babesia microti infection in Europe, a European rodent isolate (HK) and a zoonotic American isolate (GI) were studied in transmission experiments. PCR detected B. microti in the blood and spleens of infected gerbils (Meriones unguiculatus) and also in laboratory-induced infections of I. ricinus ticks. B. microti DNA was detected by PCR in all pooled samples of nymphs and the majority of adults that had fed as larvae and nymphs, respectively, on gerbils with acute infection of the European isolate, confirming that I. ricinus could serve as a vector in Europe. The American isolate, GI, proved to be equally infective for larval and nymphal I. ricinus as the HK strain, despite a very different appearance in gerbil erythrocytes. Nymphs infected with the HK and GI strains readily infected gerbils. In contrast to the finding in acute infections, ticks that fed on gerbils with chronic infections of HK and GI did not become infected. It was also found that the HK strain was not transmitted transovarially. The finding that a B. microti strain (GI) from a distant geographical region (United States) can infect and be transmitted by I. ricinus suggests that other European B. microti strains, in addition to the HK strain used here, are probably infective for I. ricinus, supporting the view that infection of humans with European B. microti may be a regular occurrence. PMID- 11923343 TI - Limited genetic diversity of recent invasive isolates of non-serotype b encapsulated Haemophilus influenzae. AB - Invasive infections caused by non-type b encapsulated Haemophilus influenzae have increased in frequency in the last decade. This change prompted us to characterize the genetic relationships of 48 recently isolated invasive H. influenzae type a (Hia), e (Hie), and f (Hif) strains by comparison of restriction digest patterns (RDPs). Recent Hia isolates exhibited moderate genetic diversity, with the majority segregating into two major clonotypes. Recent Hie and, especially, Hif strains displayed considerably restricted genetic diversity. In particular, all but one Hif strain segregated into a single clonotype, and half of these isolates had identical RDPs. These results are consistent with the hypothesis that the increased incidence of disease due to non type b encapsulated H. influenzae reflects the emergence of hypervirulent clones, especially in the case of Hif. Alternatively, it is possible that non-type b encapsulated H. influenzae strains have limited overall genetic diversity. PMID- 11923344 TI - Simultaneous differentiation and typing of Entamoeba histolytica and Entamoeba dispar. AB - Sequences corresponding to some of the polymorphic loci previously reported from Entamoeba histolytica have been detected in Entamoeba dispar. Comparison of nucleotide sequences of two loci between E. dispar strain SAW760 and E. histolytica strain HM-1:IMSS revealed significant differences in both repeat and flanking regions. The tandem repeat units varied not only in sequence but also in number and arrangement between the two species at both the loci. Using the sequences obtained, primer pairs aimed at amplifying species-specific products were designed and tested on a variety of E. histolytica and E. dispar samples. Amplification results were in complete agreement with the original species classification in all cases, and the PCR products displayed discernible size and pattern variations among the isolates. PMID- 11923345 TI - Evolution of the IS6110-based restriction fragment length polymorphism pattern during the transmission of Mycobacterium tuberculosis. AB - Interpretation of the molecular epidemiological data of Mycobacterium tuberculosis is dependent on the validity of the assumptions that have been made. It is assumed that the IS6110 banding pattern is sufficiently stable to define epidemiological events representing ongoing transmission. However, molecular epidemiological data also support the observation that the IS6110 banding pattern may change over time. Factors affecting this rate may include the nature and duration of disease in a host and the opportunity to experience different host environments during the transmission cycle. To estimate the rate of IS6110 change occurring during the process of transmission, M. tuberculosis isolates from epidemiologically linked patients were genotypically characterized by restriction fragment length polymorphism (RFLP) analysis. The identification of IS6110 banding pattern changes during ongoing transmission suggested that a rate could be estimated. IS6110 change was significantly associated with strains with >5 IS6110 elements (P = 0.013) and was not observed in low-copy-number isolates. The minimum rate of appearance of variant strains was calculated to be 0.14 variant cases per source-case per year. This data suggest that clustering of isolates based on identical RFLP patterns is expected to underestimate transmission in patients infected with high-copy-number isolates. A model based on the rate of appearance of both variant and invariant strains demonstrates that the genotypically defined population structure may change by 18.6% during the study period of approximately 6.5 years. The implications for the use of RFLP data for epidemiologic study are discussed. PMID- 11923346 TI - Mitochondrial telomeres as molecular markers for identification of the opportunistic yeast pathogen Candida parapsilosis. AB - Recent studies have demonstrated that a large number of organisms carry linear mitochondrial DNA molecules possessing specialized telomeric structures at their ends. Based on this specific structural feature of linear mitochondrial genomes, we have developed an approach for identification of the opportunistic yeast pathogen Candida parapsilosis. The strategy for identification of C. parapsilosis strains is based on PCR amplification of specific DNA sequences derived from the mitochondrial telomere region. This assay is complemented by immunodetection of a protein component of mitochondrial telomeres. The results demonstrate that mitochondrial telomeres represent specific molecular markers with potential applications in yeast diagnostics and taxonomy. PMID- 11923347 TI - Usefulness of multilocus sequence typing for characterization of clinical isolates of Candida albicans. AB - Molecular characterization of Candida albicans isolates is essential for understanding the epidemiology of nosocomial infections caused by this yeast. Here, we investigated the potential value of multilocus sequence typing (MLST) for characterizing epidemiologically related or unrelated C. albicans strains of various clinical origins. Accordingly, we sequenced the internal regions (loci) of six selected housekeeping genes of 40 C. albicans clinical isolates and 2 reference strains. In all, 68 polymorphic nucleotide sites were identified, of which 65 were found to be heterozygous in at least one isolate. Ten to 24 different genotypes were observed at the different loci, resulting, when combined, in 39 unique genotype combinations or diploid sequence types (DSTs). When MLST was applied to 26 epidemiologically unrelated isolates and the 2 reference strains, it allowed the identification of 27 independent DSTs, thus demonstrating a discriminatory power of 99.7. Using multidimensional scaling together with the minimum spanning tree method to analyze interstrain relationships, we identified six groups of genetically related isolates on the basis of bootstrap values of greater than 900. Application of MLST to 14 epidemiologically related isolates showed that those recovered from patients in the same hospital ward during the same 3 months had specific DSTs, although 73% of these isolates were genetically very close. This suggests that MLST can trace minute variations in the sequences of related isolates. Overall, MLST proved to be a highly discriminatory and stable method for unambiguous characterization of C. albicans. PMID- 11923348 TI - Epidemiology of candidemia: 3-year results from the emerging infections and the epidemiology of Iowa organisms study. AB - Bloodstream infections due to Candida species cause significant morbidity and mortality. Surveillance for candidemia is necessary to detect trends in species distribution and antifungal resistance. We performed prospective surveillance for candidemia at 16 hospitals in the State of Iowa from 1 July 1998 through 30 June 2001. Using U.S. Census Bureau and Iowa Hospital Association data to estimate a population denominator, we calculated the annual incidence of candidemia in Iowa to be 6.0 per 100,000 of population. Candida albicans was the most common species detected, but 43% of candidemias were due to species other than C. albicans. Overall, only 3% of Candida species were resistant to fluconazole. However, Candida glabrata was the most commonly isolated species other than C. albicans and demonstrated some resistance to azoles (fluconazole MIC at which 90% of the isolates tested are inhibited, 32 microg/ml; 10% resistant, 10% susceptible dose dependent). C. glabrata was more commonly isolated from older patients (P = 0.02) and caused over 25% of candidemias among persons 65 years of age or older. The investigational triazoles posaconazole, ravuconazole, and voriconazole had excellent in vitro activity overall against Candida species. C. albicans is the most important cause of candidemia and remains highly susceptible to available antifungal agents. However, C. glabrata has emerged as an important and potentially antifungal resistant cause of candidemia, particularly among the elderly. PMID- 11923349 TI - Genome scale comparison of Mycobacterium avium subsp. paratuberculosis with Mycobacterium avium subsp. avium reveals potential diagnostic sequences. AB - The genetic similarity between Mycobacterium avium subsp. paratuberculosis and other mycobacterial species has confounded the development of M. avium subsp. paratuberculosis-specific diagnostic reagents. Random shotgun sequencing of the M. avium subsp. paratuberculosis genome in our laboratories has shown >98% sequence identity with Mycobacterium avium subsp. avium in some regions. However, an in silico comparison of the largest annotated M. avium subsp. paratuberculosis contigs, totaling 2,658,271 bp, with the unfinished M. avium subsp. avium genome has revealed 27 predicted M. avium subsp. paratuberculosis coding sequences that do not align with M. avium subsp. avium sequences. BLASTP analysis of the 27 predicted coding sequences (genes) shows that 24 do not match sequences in public sequence databases, such as GenBank. These novel sequences were examined by PCR amplification with genomic DNA from eight mycobacterial species and ten independent isolates of M. avium subsp. paratuberculosis. From these analyses, 21 genes were found to be present in all M. avium subsp. paratuberculosis isolates and absent from all other mycobacterial species tested. One region of the M. avium subsp. paratuberculosis genome contains a cluster of eight genes, arranged in tandem, that is absent in other mycobacterial species. This region spans 4.4 kb and is separated from other predicted coding regions by 1,408 bp upstream and 1,092 bp downstream. The gene upstream of this eight-gene cluster has strong similarity to mycobacteriophage integrase sequences. The GC content of this 4.4 kb region is 66%, which is similar to the rest of the genome, indicating that this region was not horizontally acquired recently. Southern hybridization analysis confirmed that this gene cluster is present only in M. avium subsp. paratuberculosis. Collectively, these studies suggest that a genomics approach will help in identifying novel M. avium subsp. paratuberculosis genes as candidate diagnostic sequences. PMID- 11923350 TI - Detection and multigenic characterization of a herpesvirus associated with malignant catarrhal fever in white-tailed deer (Odocoileus virginianus) from Missouri. AB - Between 1998 and 2001, tissues from four captive white-tailed deer were observed to have histologic lesions of systemic lymphocytic vasculitis. These lesions suggested malignant catarrhal fever, although epizootic hemorrhagic disease and bluetongue were included in the differential diagnosis. Initial diagnostic efforts, including virus isolation and reverse transcription-PCR for epizootic hemorrhagic disease virus and bluetongue virus, failed to identify an etiologic agent. However, consensus primer PCR targeted to the herpesvirus DNA polymerase gene detected viral genomic DNA in each of these four cases. Nucleotide sequence analysis of the amplified product demonstrated that the detected virus was identical over the compared region to the recently described malignant catarrhal fever virus of white-tailed deer (H. Li, N. Dyer, J. Keller, and T. B. Crawford, J. Clin. Microbiol. 38:1313-1318, 2000). Additional nucleotide sequencing of both the DNA polymerase gene and DNA packaging gene followed by phylogenetic analysis solidified this newly recognized herpesvirus as a member of the Gammaherpesvirinae and suggests that this virus, along with ovine herpesvirus 2, alcelaphine herpesvirus 1, alcelaphine herpesvirus 2 and caprine herpesvirus 2, may be part of a separate clade within this subfamily. PMID- 11923351 TI - Frequency of mutations conferring resistance to nucleoside reverse transcriptase inhibitors in human immunodeficiency virus type 1-infected patients in Korea. AB - A nested PCR and direct sequencing methods were used to define human immunodeficiency virus type 1(HIV-1) reverse transcriptase codons 41 to 219 in DNA from 127 peripheral blood mononuclear cell samples obtained from 35 patients treated with nucleoside reverse transcriptase inhibitors (NRTI). The follow-up period after the initiation of NRTI therapy was 61.8 +/- 31 months (mean and standard deviation). In addition to NRTI therapy, 32 of 35 patients were simultaneously treated with Korean red ginseng. The annual decrease in the CD4(+) T-cell count over 5 years was 13.2/microl. Twenty-eight (80%) of the 35 patients had mutations conferring resistance to NRTI. The frequencies of K70R, T215S/Y/F (i.e., mutation of T at codon 215 to S, Y, or F), D67N/E, K219Q, T69N/S/A, M41L, and L210W mutations conferring resistance to zidovudine were 57.6, 36.4, 36.4, 27.2, 24.2, 21.2, and 12.1%, respectively. Mutations conferring resistance to didanosine and lamivudine were detected in 2 (L74V and M184I; 14.2%) of 11 patients tested and in 4 (M184V; 57%) of 7 patients tested, respectively. In particular, the frequency of T69N/S/A increased sharply after more than 48 months of zidovudine monotherapy. However, Q151M was not detected. As the first report on the frequency of NRTI resistance mutations in Korea, our data suggest that genotypic antiretroviral drug testing should be considered for the design of better drug regimens to improve the management of HIV-1-infected patients. PMID- 11923352 TI - Detection by reverse transcription-PCR and genetic characterization of field isolates of swine hepatitis E virus from pigs in different geographic regions of the United States. AB - Hepatitis E virus (HEV) is an important public health concern in many developing countries. HEV is also endemic in some industrialized counties, including the United States. With our recent discovery of swine HEV in pigs that is genetically closely related to human HEV, hepatitis E is now considered a zoonotic disease. Human strains of HEV are genetically heterogenic. So far in the United States, only one strain of swine HEV has been identified and characterized from a pig. To determine the extent of genetic variations and the nature of swine HEV infections in U.S. pigs, we developed a universal reverse transcription-PCR (RT-PCR) assay that is capable of detecting genetically divergent strains of HEV. By using this universal RT-PCR assay, we tested fecal and serum samples of pigs of 2 to 4 months of age from 37 different U.S. swine farms for the presence of swine HEV RNA. Thirty-four of the 96 pigs (35%) and 20 of the 37 swine herds (54%) tested were positive for swine HEV RNA. The sequences of a 348-bp region within the ORF2 gene of 27 swine HEV isolates from different geographic regions were determined. Sequence analyses revealed that the 27 U.S. swine HEV isolates shared 88 to 100% nucleotide sequence identities with each other and 89 to 98% identities with the prototype U.S. strain of swine HEV. These U.S. swine HEV isolates are only distantly related to the Taiwanese strains of swine HEV, with about 74 to 78% nucleotide sequence identities; to most known human strains of HEV worldwide, with <79% sequence identities; and to avian HEV, with 54 to 56% sequence identities. Phylogenetic analysis showed that all the U.S. swine HEV isolates identified in this study clustered in the same genotype with the prototype U.S. swine HEV and the two U.S. strains of human HEV. The data from this study indicated that swine HEV is widespread and enzoonotic in U.S. swine herds and that, as is with human HEV, swine HEV isolates from different geographic regions of the world are also genetically heterogenic. These data further raise potential concerns for zoonosis, xenozoonosis, and food safety. PMID- 11923353 TI - rpoB gene sequence-based identification of Staphylococcus species. AB - The complete sequence of rpoB, the gene encoding the beta subunit of RNA polymerase was determined for Staphylococcus saccharolyticus, Staphylococcus lugdunensis, S taphylococcus caprae, and Staphylococcus intermedius and partial sequences were obtained for an additional 27 Staphylococcus species. The complete rpoB sequences varied in length from 3,452 to 3,845 bp and had a 36.8 to 39.2% GC content. The partial sequences had 71.6 to 93.6% interspecies homology and exhibited a 0.08 to 0.8% intraspecific divergence. With a few exceptions, the phylogenetic relationships inferred from the partial rpoB sequences were in agreement with those previously derived from DNA-DNA hybridization studies and analyses of 16S ribosomal DNA gene sequences and partial HSP60 gene sequences. The staphylococcal rpoB sequence database we established enabled us to develop a molecular method for identifying Staphylococcus isolates by PCR followed by direct sequencing of the 751-bp amplicon. In blind tests, this method correctly identified 10 Staphylococcus isolates, and no positive results were obtained with 10 non-Staphylococcus gram-positive and gram-negative bacterial isolates. We propose partial sequencing of the rpoB gene as a new tool for the accurate identification of Staphylococcus isolates. PMID- 11923354 TI - Detection of Mycoplasma pneumoniae in spiked clinical samples by nucleic acid sequence-based amplification. AB - Isothermal nucleic acid sequence-based amplification (NASBA) was applied to the detection of Mycoplasma pneumoniae. M. pneumoniae RNA prepared from a plasmid construct was used to assess the sensitivity of the assay, and an internal control for the detection of inhibitors was constructed. The sensitivity of the NASBA assay was 10 molecules of wild-type M. pneumoniae RNA generated in vitro and 5 color-changing units (CCU) of M. pneumoniae. An appropriate specimen preparation procedure was developed: after protease treatment of the respiratory specimens, guanidine thiocyanate lysis solution (4.7 M guanidine thiocyanate [Sigma-Aldrich NV], 46 mM Tris-HCl [Merck, Darmstadt, Germany], 20 mM EDTA [Sigma Aldrich NV], 1.2% [wt/vol] Triton X-100 [Sigma-Aldrich NV], pH 6.2.) was added. With spiked throats, nasopharyngeal aspirates, bronchoalveolar lavage specimens, and sputum specimens, the sensitivity of the NASBA assay in the presence of the internal control was 2 x 10(4) molecules of in vitro-generated RNA or 5 CCU of M. pneumoniae. The sensitivity of the NASBA assay was comparable to that of a PCR targeted to the P1 adhesin gene. Fifteen clinical specimens positive for M. pneumoniae by PCR were also positive by NASBA. These results indicate that the sensitivity of detection of M. pneumoniae in spiked respiratory samples by NASBA is high. Together with the use of the internal control, the assay merits evaluation as a diagnostic tool. PMID- 11923355 TI - Identification of medically relevant Nocardia species with an abbreviated battery of tests. AB - Identification of Nocardia to the species level is useful for predicting antimicrobial susceptibility patterns and defining the pathogenicity and geographic distribution of these organisms. We sought to develop an identification method which was accurate, timely, and employed tests which would be readily available in most clinical laboratories. We evaluated the API 20C AUX yeast identification system as well as several biochemical tests and Kirby-Bauer susceptibility patterns for the identification of 75 isolates encompassing the 8 medically relevant Nocardia species. There were few biochemical reactions that were sufficiently unique for species identification; of note, N. nova were positive for arylsulfatase, N. farcinica were positive for opacification of Middlebrook 7H11 agar, and N. brasiliensis and N. pseudobrasiliensis were the only species capable of liquefying gelatin. API 20C sugar assimilation patterns were unique for N. transvalensis, N. asteroides IV, and N. brevicatena. There was overlap among the assimilation patterns for the other species. Species-specific patterns of susceptibility to gentamicin, tobramycin, amikacin, and erythromycin were obtained for N. nova, N. farcinica, and N. brevicatena, while there was overlap among the susceptibility patterns for the other isolates. No single method could identify all Nocardia isolates to the species level; therefore, a combination of methods was necessary. An algorithm utilizing antibiotic susceptibility patterns, citrate utilization, acetamide utilization, and assimilation of inositol and adonitol accurately identified all isolates. The algorithm was expanded to include infrequent drug susceptibility patterns which have been reported in the literature but which were not seen in this study. PMID- 11923357 TI - New yeast species, Malassezia dermatis, isolated from patients with atopic dermatitis. AB - Malassezia species are considered to be one of the exacerbating factors in atopic dermatitis (AD). During examination of the cutaneous colonization of Malassezia species in AD patients, we found a new species on the surface of the patients' skin. Analysis of ribosomal DNA sequences suggested that the isolates belonged to the genus MALASSEZIA: They did not grow in Sabouraud dextrose agar but utilized specific concentrations of Tween 20, 40, 60, and 80 as a lipid source. Thus, we concluded that our isolates were new members of the genus Malassezia and propose the name Malassezia dermatis sp. nov. for these isolates. PMID- 11923356 TI - Genetic and phenotypic differences between Legionella pneumophila strains. AB - Legionnaires' disease is a potentially lethal pneumonia that is primarily due to infection by the species Legionella pneumophila, although more than 40 other species are known. Certain L. pneumophila subgroups, particularly serogroup 1, are associated with the majority of the epidemics. The genetic bases for these differences in virulence have not been determined. Three strains, AA100, JR32, and Lp01, have been used in many molecular pathogenesis studies of L. pneumophila. We found genetic differences between these strains by PCR and Southern analyses that may be related to their ability to cause disease. We also examined the distribution of these genetic loci in clinical and environmental isolates of Legionella and found a correlation between the presence of two of these loci, rtxA and lvh, and the ability to cause disease in humans. Examination of the interactions of these strains with host cells suggested that they differ in important phenotypic characteristics including adherence, entry, and intracellular replication. Furthermore, in the mouse model of infection they display differing levels of replication in lungs. These studies emphasize the importance of further investigation into the genetic makeup of these strains, which is likely to lead to the identification of additional factors involved in Legionella pathogenesis. PMID- 11923358 TI - Detection of mammalian reovirus RNA by using reverse transcription-PCR: sequence diversity within the lambda3-encoding L1 gene. AB - Reoviruses infect virtually all mammalian species, and infection of humans is associated with mild gastrointestinal or upper respiratory illnesses. To improve reovirus detection strategies, we developed a reverse transcription-PCR technique to amplify a fragment of the reovirus L1 gene segment. This assay was capable of detecting 44 of 44 reovirus field isolate strains and was sufficiently sensitive to detect nearly a single viral particle (1.16 +/- 0.13) per PCR of prototype strain type 3 Dearing. Pairwise comparisons of the 44 partial L1 gene sequences revealed that nucleotide variability ranged from 0 to 24.7%, with most of the nucleotide polymorphism occurring at synonymous positions. Phylogenetic trees generated from amplified L1 gene sequences suggest that multiple alleles of the L1 gene cocirculate in nature and that genetic diversity of the L1 gene is largely independent of the host species, geographic locale, or date of isolation. Phylogenetic trees constructed from the L1 gene sequences are distinct from those constructed from the four reovirus S-class gene segments, which supports the hypothesis that reovirus gene segments reassort in nature. This study establishes a new sensitive and specific technique for the identification of mammalian reoviruses and enhances our understanding of reovirus evolution. PMID- 11923359 TI - Clonal spread of Staphylococcus aureus heterogeneously resistant to vancomycin in a university hospital in Korea. AB - Since vancomycin-intermediate Staphylococcus aureus (VISA) was first reported in Japan in 1997, there has been great concern that heterogeneous vancomycin intermediate S. aureus (hetero-VISA) is the putative precursor of VISA. To investigate the prevalence, clinical significance, and molecular epidemiology of S. aureus with reduced susceptibility to vancomycin, all consecutive isolates of S. aureus isolated from clinical specimens from December 1998 to August 1999 at Asan Medical Center were screened for VISA and hetero-VISA by using brain heart infusion agar containing 4 microg of vancomycin/ml. Screen-positive isolates were confirmed by susceptibility testing and population analysis of subpopulations with reduced susceptibility to vancomycin. The isolates confirmed as hetero-VISA were typed by pulsed-field gel electrophoresis (PFGE). Medical records were reviewed to evaluate the clinical significance and risk factors for the acquisition of hetero-VISA. Of the 4,483 isolates that were tested, 53 were screen positive; no VISA was detected, but 24 isolates (0.54%) from 22 patients were hetero-VISA. All but two strains appeared to be clones of the Korean VISA strain, AMC11094, in the PFGE analysis. A total of 18 patients were in intensive care units, and 16 underwent major surgeries during the same admission. Only 10 of the 22 patients had previous methicillin-resistant S. aureus infections and 11 had previous vancomycin or teicoplanin therapy. Only 7 of the 22 patients from whom hetero-VISA strains were isolated were infected, and the remaining 15 patients were colonized. All seven infected patients were successfully treated with vancomycin. These results suggest that hetero-VISA can be treated with vancomycin, but the spread of hetero-VISA clonal to VISA is of concern, since many believe that VISA can arise from hetero-VISA, although this phenomenon was not observed in this study. PMID- 11923360 TI - Catheter-related candidemia caused by Candida lipolytica in a patient receiving allogeneic bone marrow transplantation. AB - Candida lipolytica was recovered from the blood and the central venous catheter in a patient receiving allogeneic bone marrow transplantation. Two C. lipolytica strains from different geographical areas and the ATCC 9773 strain of C. lipolytica were used as controls. C. lipolytica was identified by standard methods. MICs indicated antifungal susceptibilities to amphotericin B, fluconazole, and itraconazole for all strains. In vitro testing and scanning electron microscopy showed that C. lipolytica was capable of producing large amounts of viscid slime material in glucose-containing solution, likely responsible for the ability of the yeast to adhere to catheter surfaces. Restriction fragment length polymorphisms revealed an identical profile for all clinical isolates, unrelated to those observed for the control strains. This finding suggested the absence of microevolutionary changes in the population of the infecting strain, despite the length of the sepsis and the potential selective pressure of amphotericin B, which had been administered to the patient for about 20 days. The genomic differences that emerged between the isolates and the control strains were indicative of a certain degree of genetic diversity between C. lipolytica isolates from different geographical areas. PMID- 11923361 TI - Timing of inoculation of the pouch makes no difference in increased detection of Trichomonas vaginalis by the InPouch TV method. AB - The InPouch TV is a method which combines a wet preparation and a culture method to detect Trichomonas vaginalis. The top portion of the InPouch TV essentially functions as a slide to be examined under the microscope. If the initial examination is negative, the specimen is pushed down into the bottom pouch, which serves as a broth for cultivation. The issue of timing has not been specifically addressed for optimal processing. To assess the effect of timing on the inoculation of the bottom pouch, we conducted a study designed to determine which procedure had better sensitivity, that of delaying inoculation of the bottom pouch until the initial examination on the top pouch is performed (method A) or that of immediately inoculating the bottom pouch (method B). In addition, we compared the sensitivity of the InPouch TV to that of the traditional wet mount. Fifty of 498 specimens were positive. Methods A and B had identical results: 31 specimens were initially positive regardless of transit time, and incubation yielded another 19 positives. The wet preparation detected 36 positive specimens. The sensitivities of the methods were 100% for the InPouch TV (including examination on receipt and after incubation) and 72% for the traditional wet mount. In conclusion, the InPouch TV method is more sensitive than the traditional method and no detectable differences were observed with timing of the inoculation of the top or bottom pouch. PMID- 11923362 TI - Detection of a human rotavirus with G12 and P[9] specificity in Thailand. AB - G12 rotavirus has not been detected anywhere in the world since the first detection of a human strain, L26 (G12, P1B[4]), in the Philippines in 1990. In this study, we isolated a human rotavirus (strain T152) with a VP7 of G12 specificity from the stool of an 11-month-old diarrheic patient in Thailand. The strain T152 exhibited a long RNA pattern and subgroup I specificity. In the comparison of the nucleotide and amino acid sequences of the VP7 gene of strain T152 with those of rotaviruses with different G type specificities, strain T152 showed the highest identity, 90.9 and 93.9%, respectively, to G12 prototype strain L26. In contrast, the VP4 gene of strain T152 showed the highest identity with P[9] specificity of human strains K8 and AU-1 and feline strains Cat2 and FRV-1, with homologies of 89.3 to 90.6% at the nucleotide level and 93.9 to 95.6% at the amino acid level. Thus, strain T152 was found to be a natural reassortant strain with G12 and P[9] specificities. PMID- 11923363 TI - Reactivity of convalescent-phase hemolytic-uremic syndrome patient sera with the megaplasmid-encoded TagA protein of Shiga toxigenic Escherichia coli O157. AB - A cosmid library of Shiga toxigenic Escherichia coli (STEC) O157:H7 strain EDL933 DNA was screened for clones capable of reacting with convalescent-phase serum from a patient with hemolytic-uremic syndrome (HUS), in an attempt to identify candidate virulence genes. One of the immunoreactive clones contained a portion of the large plasmid pO157, and the immunoreactive gene product was identified as TagA. The function of this 898-amino-acid protein is unknown, but it exhibits 42% amino acid sequence identity and 63% similarity to a 312-amino-acid region of a ToxR-regulated lipoprotein of Vibrio cholerae. Antibodies to E. coli O157 TagA were detected in sera from other HUS patients with O157 STEC infection but not in those from patients whose illnesses were caused by other STEC types or in healthy controls. These data demonstrate that TagA is expressed in vivo and provide circumstantial evidence for a role in the pathogenesis of the disease. The tagA gene is present only in STEC strains belonging to serogroup O157, and so antibodies to TagA are a potentially useful serological marker for infections due to such strains. PMID- 11923364 TI - Immunoglobulin M (IgM)-glycoinositolphospholipid enzyme-linked immunosorbent assay: an immunoenzymatic assay for discrimination between patients with acute toxoplasmosis and those with persistent parasite-specific IgM antibodies. AB - In the present study we developed an enzyme-linked immunosorbent assay (ELISA) to measure immunoglobulin M (IgM) specific for glycoinositolphospholipids (GIPL) derived from tachyzoite membrane (IgM-GIPL ELISA). The sensitivity and specificity of the assay were compared with those of commercially available Toxoplasma-specific IgM serological tests, namely, immunofluorescence assay (IFA) with fixed tachyzoites and capture ELISA employing tachyzoite extracts. Our results show that all patients with acute toxoplasmosis, as determined by clinical data and conventional serological tests, were also positive by the IgM GIPL ELISA. Interestingly, many patients that were classified as indeterminate, who had IgG with high avidity but positive results in the IgM-specific IFA and capture ELISA, were negative by the IgM-GIPL ELISA. Finally, we tested the sera from patients with rheumatoid arthritis and various parasitic infections and found no evidence of false positives in the IgM-GIPL ELISA. PMID- 11923365 TI - In vitro activity of nystatin compared with those of liposomal nystatin, amphotericin B, and fluconazole against clinical Candida isolates. AB - We investigated the in vitro activity of nystatin and liposomal nystatin against 103 Candida isolates to determine the effect of both time and medium on MICs. We also compared the nystatin MICs with those of amphotericin B and fluconazole. Testing was performed in accordance with the National Committee for Clinical Laboratory Standards M27-A microdilution methodology with RPMI 1640, RPMI 1640 supplemented with glucose to 2% (RPMI-2), and antibiotic medium 3 supplemented with glucose to 2% (AM3). While nystatin MICs were similar to or slightly lower than liposomal nystatin MICs in RPMI 1640 and RPMI-2, they were markedly higher than liposomal nystatin MICs in AM3. Use of AM3 and determination of the MIC after 24 h of incubation provided a slightly wider range of liposomal nystatin MICs (0.06 to >16 microg/ml). Under these conditions, the MICs at which 90% of isolates were inhibited of nystatin and liposomal nystatin were 2 and 1 microg/ml, respectively. Nystatin and liposomal nystatin in general showed good activity against all Candida spp. tested. Although the MICs of nystatin and liposomal nystatin tended to rise in parallel with the amphotericin B MICs, nystatin and liposomal nystatin MICs of 1 to 2 and 0.5 to 1 microg/ml, respectively, were obtained for seven and six, respectively, of nine isolates for which amphotericin B MICs were >or=0.25 microg/ml. No correlation between fluconazole and nystatin or liposomal nystatin MICs was observed. As amphotericin B MICs of >or=0.25 microg/ml correlate with in vitro resistance, these results suggest that liposomal nystatin might have activity against some amphotericin B resistant isolates. In vivo testing in animal models is required for clarification of this issue. PMID- 11923366 TI - Rapid and sensitive oligonucleotide ligation assay for detection of mutations in human immunodeficiency virus type 1 associated with high-level resistance to protease inhibitors. AB - A sensitive, specific, and high-throughput oligonucleotide ligation assay (OLA) for the detection of genotypic human immunodeficiency virus type 1 (HIV-1) resistance to Food and Drug Administration-approved protease inhibitors was developed and evaluated. This ligation-based assay uses differentially modified oligonucleotides specific for wild-type or mutant sequences, allowing sensitive and simple detection of both genotypes in a single well of a microtiter plate. Oligonucleotides were designed to detect primary mutations associated with high level resistance to amprenavir, nelfinavir, indinavir, ritonavir, saquinavir, and lopinavir, including amino acid substitutions D30N, I50V, V82A/S/T, I84V, N88D, and L90M. Plasma HIV-1 RNA from 54 infected patients was amplified by reverse transcription-PCR and sequenced by using dideoxynucleotide chain terminators for evaluation of mutations associated with drug resistance. These same amplicons were genotyped by the OLA at positions 30, 50, 82, 88, 84, and 90 for a total of 312 codons. The sensitivity of detection of drug-resistant genotypes was 96.7% (87 of 90 mutant codons) in the OLA compared to 92.2% (83 of 90) in consensus sequencing, presumably due to the increased sensitivity of the OLA. The OLA detected genetic subpopulations more often than sequencing, detecting 30 mixtures of mutant and wild-type sequences and two mixtures of drug-resistant sequences compared to 15 detected by DNA sequencing. Reproducible and semiquantitative detection of the mutant and the wild-type genomes by the OLA was observed by analysis of wild-type and mutant plasmid mixtures containing as little as 5% of either genotype in a background of the opposite genome. This rapid, simple, economical, and highly sensitive assay provides a practical alternative to dideoxy sequencing for genotypic evaluation of HIV-1 resistance to antiretrovirals. PMID- 11923367 TI - Evaluation of a new combined antigen and antibody human immunodeficiency virus screening assay, VIDAS HIV DUO Ultra. AB - Combined antigen and antibody screening (fourth-generation) assays reduce the diagnostic window period between the time of human immunodeficiency virus (HIV) infection and laboratory diagnosis by 4 days, on average, in comparison to antibody-only (third generation) enzyme immunoassays (EIAs). The aim of the present study was to assess whether the new VIDAS HIV DUO Ultra (Biomerieux, Marcy-l'Etoile, France) showed an improved sensitivity and specificity in comparison to licensed fourth-generation assays. A total of 16 seroconversion panels, 15 cell culture supernatants infected with different HIV type 1 (HIV-1) subtypes, and 257 potentially cross-reactive serum samples were tested with VIDAS DUO HIV Ultra, Genscreen Plus HIV Ag-Ab, Enzygnost HIV Integral, Enzymun-Test HIV Combi, Genscreen HIV 1/2, version 2 (third-generation EIA), and Genetic Systems HIV-1 Ag EIA (p24 antigen assay). VIDAS HIV DUO Ultra showed a comparable sensitivity to the single p24 antigen assay in seroconversion panels and a dilution series of virus lysates. The diagnostic window was reduced with VIDAS HIV DUO Ultra by 3.82 days, on average, in comparison with the fourth-generation assay with the lowest sensitivity of the antigen detection module. HIV-1 infection was detected 5.88 days earlier than with third-generation EIA. The mean time delay between reverse transcription-PCR and VIDAS HIV DUO Ultra was only 2.31 days. The specificity of fourth-generation assays after retesting ranged between 98.1 and 100%. In conclusion, VIDAS HIV DUO Ultra can replace single antigen screening for laboratory diagnosis and screening of HIV infection in blood donors. There was no evidence for a second diagnostic window due to impaired sensitivity of the antibody detection module of all the fourth generation EIAs evaluated in the present study. The specificity after initial and/or repeated testing of VIDAS HIV DUO Ultra was equivalent to that of a third generation assay. PMID- 11923368 TI - Identification of a novel methicillin-resistant Staphylococcus aureus epidemic clone in Cordoba, Argentina, involved in nosocomial infections. AB - Infections caused by methicillin-resistant Staphylococcus aureus (MRSA) are increasingly a main health concern worldwide for hospitalized patients. In addition, the prevalence of community-acquired infection has risen continuously during the last few years. Some MRSA clones spread easier than others within the hospital environment and therefore are frequently implicated in outbreaks. Thus, the spread of a unique epidemic multiresistant clone, the so-called South American clone, is the main cause of nosocomial infections produced by this bacterium in Brazil and in some regions of Argentina, Chile, and Uruguay. In the present work we describe the identification of a novel clone of MRSA that is involved in nosocomial infections and that shows a prevalence as high as that for the South American clone. A total of 53 consecutive single-patient MRSA isolates were recovered during a 3-month period (May to July 1999) from six different hospitals (955 beds) in Cordoba. The isolates were initially typed according the antibiotic resistance and phage susceptibility patterns, followed by genotyping using pulsed-field gel electrophoresis (PFGE). PFGE analysis of the 53 MRSA isolates revealed six major types (A to F) and 25 subtypes. The B-type DNA pattern was indistinguishable from that of the South American epidemic clone observed in 34% of the isolates. A novel highly prevalent clone, showing the A type DNA pattern and representing 38% of the isolates, was also identified. Moreover, the most frequent subtype of the A clonal family triggered an outbreak in a hospital 2 months later, further confirming its epidemic feature. PMID- 11923369 TI - Rapid detection of Shiga toxin-producing bacteria in feces by multiplex PCR with molecular beacons on the smart cycler. AB - We have developed a rapid (1-h) real-time fluorescence-based PCR assay with the Smart Cycler thermal cycler (Cepheid, Sunnyvale, Calif.) for the detection of Shiga toxin-producing Escherichia coli (STEC), as well as other Shiga toxin producing bacteria. Based on multiple-sequence alignments, we have designed two pairs of PCR primers that efficiently amplify all variants of the Shiga toxin genes stx(1) and stx(2), respectively. These primer pairs were combined for use in a multiplex assay. Two molecular beacons bearing different fluorophores were used as internal probes specific for each amplicon. Assays performed with purified genomic DNA from a variety of STEC strains (n = 23) from diverse geographic locations showed analytical sensitivities of about 10 genome copies per PCR. Non-STEC strains (n = 20) were also tested, and no amplification was observed. The PCR results correlated perfectly with the phenotypic characterization of toxin production in both STEC and non-STEC strains, thereby confirming the specificity of the assay. The assay was validated by testing 38 fecal samples obtained from 27 patients. Of these samples, 26 were PCR positive for stx(1) and/or stx(2). Compared with the culture results, both the sensitivity and the negative predictive value were 100%. The specificity was 92%, and the positive predictive value was 96%. Moreover, this assay detected STEC from a sample in which the STEC concentration was at the limit of detection of the conventional culture methods and from a sample in which STEC was not detected by the conventional culture methods. This real-time PCR assay is simple, rapid, sensitive, and specific and allows detection of all Shiga toxin-producing bacteria directly from fecal samples, irrespective of their serotypes. PMID- 11923370 TI - Identification, characterization, and distribution of a Shiga toxin 1 gene variant (stx(1c)) in Escherichia coli strains isolated from humans. AB - By using sequence analysis of Shiga toxin 1 (Stx 1) genes from human and ovine Stx-producing Escherichia coli (STEC) strains, we identified an Stx1 variant in STEC of human origin that was identical to the Stx1 variant from ovine STEC, but demonstrated only 97.1 and 96.6% amino acid sequence identity in its A and B subunits, respectively, to the Stx1 encoded by bacteriophage 933J. We designated this variant "Stx1c" and developed stxB(1) restriction fragment length polymorphism and stx(1c)-specific PCR strategies to determine the frequency and distribution of stx(1c) among 212 STEC strains isolated from humans. stx(1c) was identified in 36 (17.0%) of 212 STEC strains, 19 of which originated from asymptomatic subjects and 16 of which were from patients with uncomplicated diarrhea. stx(1c) was most frequently (in 23 STEC strains [63.9%]) associated with stx(2d), but 12 (33.3%) of the 36 STEC strains possessed stx(1c) only. A single STEC strain possessed stx(1c) together with stx(2) and was isolated from a patient with hemolytic-uremic syndrome. All 36 stx(1c)-positive STEC strains were eae negative and belonged to 10 different serogroups, none of which was O157, O26, O103, O111, or O145. Stx1c was produced by all stx(1c)-containing STEC strains, but reacted weakly with a commercial immunoassay. We conclude that STEC strains harboring the stx(1c) variant account for a significant proportion of human STEC isolates. The procedures developed in this study now allow the determination of the frequency of STEC strains harboring stx(1c) among clinical STEC isolates and their association with human disease in prospective studies. PMID- 11923371 TI - Evaluation of an automated sample preparation protocol for quantitative detection of hepatitis C virus RNA. AB - The COBAS AMPLIPREP instrument for automated sample preparation has recently been introduced. In this study, the COBAS AMPLIPREP/COBAS AMPLICOR HCV MONITOR test, which includes this new molecular device, was evaluated and compared to the COBAS AMPLICOR HCV MONITOR test, which includes a manual extraction protocol. Interassay and intra-assay variation, precision, and linearity were determined, and a total of 130 clinical specimens were investigated. For determination of interassay variation, coefficients of variation were found to be between 9 and 59% for the COBAS AMPLIPREP/COBAS AMPLICOR HCV MONITOR test and between 13 and 69% for the COBAS AMPLICOR HCV MONITOR test. For determination of intra-assay variation, coefficients of variation were found to be between 7 and 13% for the COBAS AMPLIPREP/COBAS AMPLICOR HCV MONITOR test and between 8 and 16% for the COBAS AMPLICOR HCV MONITOR test. When precision of the COBAS AMPLIPREP/COBAS AMPLICOR HCV MONITOR test was tested, all results were found to be within +/-0.5 log of the expected results. Determination of linearity resulted in a quasilinear curve over 3 logs. When clinical samples were tested with the COBAS AMPLIPREP/COBAS AMPLICOR HCV MONITOR test and compared with the COBAS AMPLICOR HCV MONITOR test, all results were found within +/-0.5 log. In conclusion, the assay, which included the new molecular device, proved to be suitable for the routine molecular laboratory. It was found to be laborsaving and easy to handle. PMID- 11923372 TI - Quantitative detection of Mycoplasma genitalium from first-pass urine of men with urethritis and asymptomatic men by real-time PCR. AB - We developed a TaqMan-based real-time PCR assay for quantifying Mycoplasma genitalium. This assay is able to specifically quantify concentrations of the M. genitalium 16S rRNA gene ranging from 10(7) to 10 copies/reaction. Using the TaqMan assay, we quantified the M. genitalium 16S rRNA gene in first-pass urine of men with urethritis and asymptomatic men who were positive for M. genitalium by PCR- and phylogeny-based assay. Of 130 men with gonococcal urethritis (GU), five were positive for M. genitalium. The mycoplasma load for each specimen was <5 x 10 copies/ml. Of 84 men with chlamydial non-GU (CNGU), seven were positive for M. genitalium. One man had an M. genitalium load of <5 x 10 copies/ml, and six men had loads ranging from 1.1 x 10(7) to 2.7 x 10(2) copies/ml. Of 86 men with nonchlamydial NGU (NCNGU), 17 were positive for M. genitalium. The mycoplasma loads for these men ranged from 3.3 x 10(6) to 2.3 x 10(2) copies/ml. Of 76 asymptomatic men, only two were positive for M. genitalium. For these men, the loads were 2 x 10(2) and <5 x 10 copies/ml. The patients with NGU had significantly higher concentrations of M. genitalium in their first-pass urine than did men with GU (P < 0.01) or asymptomatic men (P < 0.05). In addition, M. genitalium loads were significantly higher in men with NCNGU than those in asymptomatic men (P < 0.05). The quantitative assessment of M. genitalium loads by the TaqMan assay will provide useful information for understanding the pathogenicity of this mycoplasma in the urogenital tract. PMID- 11923373 TI - Dynamics of dissemination and outer surface protein expression of different European Borrelia burgdorferi sensu lato strains in artificially infected Ixodes ricinus nymphs. AB - Unfed Ixodes ricinus nymphs were infected with eight different strains and clones of Borrelia afzelii and B. garinii by capillary feeding. Except one B. afzelii clone, all expressed OspC in culture. Tick midguts and salivary glands were investigated at different time intervals for the presence of borreliae and for OspA and OspC phenotypes by immunofluorescence with simultaneous staining of OspA and OspC with monoclonal antibodies. Both species were transmittable to I. ricinus. All OspC-expressing strains and clones were able to disseminate into the salivary glands. In contrast, the OspC-negative B. afzelii clone was not detectable in the salivary glands, an indication that OspC plays an important role in dissemination. OspA-positive borreliae prevailed in the midgut. OspC positives were more frequent in the salivary glands than in the midgut. Notably, simultaneously OspA- and OspC-negative borreliae were detected in both organs. Kinetics of dissemination varied with the strains. The OspC-positive B. afzelii clone and all B. garinii OspA type 4 strains were detectable in the salivary glands right after feeding, while one B. garinii OspA type 6 strain invaded the salivary glands with a delay of 24 h. These findings support the hypothesis that OspA is abundantly expressed in unfed ticks while upregulation of OspC is also a prerequisite for dissemination in the vector for the Eurasian species B. afzelii and B. garinii. However, we found strain-specific dynamics of Osp expression and strain-specific kinetics of systemic infection in the vector tick and it appears that additional factors are involved in the initiation and regulation of the dissemination process. PMID- 11923374 TI - Evaluation of eight rapid screening tests for acute leptospirosis in Hawaii. AB - Leptospirosis is a major public health problem throughout the world. Clinical recognition of leptospirosis is challenging, and the definitive serologic diagnostic assay, the microscopic agglutination test, is time-consuming and difficult to conduct. Various serologic screening tests have been developed, but their performance among ill persons in the United States has not been established. Eight screening tests were compared using 379 serum samples obtained in 1998 and 1999 from a series of 236 patients (33 with confirmed infection). The median number of days between illness onset and specimen collection was 9. The overall sensitivity, by specimen, for each test was as follows: indirect hemagglutination assay (MRL Diagnostics, Cypress, Calif.), 29%; INDX Leptospira Dip-S-Tick (PanBio InDx, Inc., Baltimore, Md.), 52%; Biognost IgM IFA test (Bios GmbH Labordiagnostik, Grafelfing, Germany), 40%; Biolisa IgM ELISA (Bios GmbH, Labordiagnostik), 48%; Leptospira IgM ELISA (PanBio Pty Ltd., Brisbane, Australia), 36%; SERION ELISA classic Leptospira (Institut Virion*Serion GmbH, Wurzburg, Germany), 48%; LEPTO Dipstick(Organon-Teknika, Ltd., Amsterdam, The Netherlands), 34%; Biosave latex agglutination test (LATEX; Bios GmbH Labordiagnostik), 86%. Test specificity ranged from 85 to 100% among all tests except LATEX, for which the specificity was significantly lower, at 10%. Test sensitivity was particularly low (<25%) for all tests (except LATEX) on specimens collected during the first week of illness. This is the most comprehensive field trial of leptospirosis screening tests reported to date. The data indicate that immunoglobulin M detection tests have limited utility for diagnosing leptospirosis during the initial evaluation of patients seen in Hawaii, a time when important therapeutic decisions are made. Improved leptospirosis screening tests are needed. PMID- 11923375 TI - Cloning of a truncated Babesia equi gene encoding an 82-kilodalton protein and its potential use in an enzyme-linked immunosorbent assay. AB - To isolate Babesia equi genes encoding immunodominant proteins, a cDNA expression library prepared from B. equi mRNA was immunoscreened with B. equi-infected horse serum. Eighteen positive cDNA clones were obtained, and the clone that showed the strongest immunoreactivity, designated Be82, was further characterized. The Be82 gene consisted of 1,953 bp and contained a partial open reading frame lacking the 5'-terminal sequence. As shown by Western blot analyses, immune sera from mice intraperitoneally injected with the Be82 gene product recognized the 82- and 52 kDa proteins of B. equi but not those of Babesia caballi. The glutathione S transferase fusion protein expressed in Escherichia coli that was purified and used as the antigen in the enzyme-linked immunosorbent assay reacted specifically with B. equi-infected horse sera. These results suggest that the Be82 gene product is a potential diagnostic antigen candidate in the detection of B. equi infection in horses that will be useful both in the performance of epidemiological studies and in the granting of quarantine passes. PMID- 11923376 TI - Identification of the Brucella melitensis vaccine strain Rev.1 in animals and humans in Israel by PCR analysis of the PstI site polymorphism of its omp2 gene. AB - Adverse effects of strain persistence and secretion in milk have been encountered with the Brucella melitensis vaccine strain Rev.1. Field isolates obtained from vaccinated animals and from a human resembled the vaccine strain Rev.1 by conventional bacteriological tests. The lack of a specific molecular marker that could specifically characterize the commercial vaccine strain prevented confirmation of the homology of the Rev.1-like field isolates to the vaccine strain. The composition of the omp2 locus from two gene copies with differences in their PstI restriction endonuclease sites was used to establish an epidemiologic fingerprint for the omp2 gene in the Rev.1 vaccine strain. Primers designed to amplify DNA sequences that overlap the PstI site revealed a single 282-bp DNA band common to all Brucella spp. Agarose gel electrophoresis of the PstI digests of the PCR products from strains 16M and the vaccine strain Rev.1 revealed a distinctive profile that included three bands: one band for the intact 282-bp fragment amplified from omp2a and two bands resulting from the digestion of the amplified omp2b gene fragment, 238- and 44-bp DNA fragments, respectively. Amplified fragments of 37 Rev.1-like isolates, including 2 human isolates, also exhibited this pattern. In contrast, DNA digests of all other Israeli field isolates, including atypical B. melitensis biotype 1 and representatives of the biotype 2 and 3 isolates, produced two bands of 238 and 44 bp, respectively, corresponding with the digestion of both omp2a and omp2b genes. This method facilitates identification of the Rev.1 vaccine strain in both animals and humans in Israel. PMID- 11923378 TI - Identification of two phylogenetically related organisms from feces by PCR for detection of Salmonella spp. AB - Two previously reported PCR methods were evaluated to determine whether they are as sensitive and specific as conventional culture methods in detecting Salmonella spp. from feces. Bovine and equine feces were enriched overnight in brain heart infusion broth and assayed using PCR methods and primer sets described by other investigators. A total of 774 fecal specimens were tested using a primer set (invE-A primer set) that amplifies a region spanning the invasin E and A genes of Salmonella enterica serovar Typhimurium. A subset of these fecal specimens (306 of the 774 total) were tested using primers (hisJ primer set) that amplify a portion of the histidine transport J gene. The PCR required 24 h to obtain results, whereas it took 5 to 7 days to identify Salmonella spp. by culture. PCR detection of Salmonella spp. using the hisJ primers and the invE-A primers had a sensitivity of 93.3 and 80%, respectively, and a specificity of 85.6 and 98.6%, respectively, compared with bacterial culture. Amplification of 42 culture negative fecal specimens (of 306 total specimens) generated a DNA fragment that corresponded to the molecular weight of the amplified hisJ gene. The hisJ generated amplicons from six culture-negative and six culture-positive specimens were sequenced and analyzed using DNA sequence alignment and phylogenetic analysis software. A neighbor-joining dendrogram of the DNA sequences of both sets of hisJ amplicons revealed two distinct groups-one group of amplicons from culture-positive specimens identical to the hisJ gene of S. enterica serovar Typhimurium and a second group of amplicons from culture-negative specimens that were more closely related to hisJ of S. enterica serovar Typhimurium than to other hisJ sequences present in nucleotide databases. PMID- 11923377 TI - Comparison of gyrA mutations, cyclohexane resistance, and the presence of class I integrons in Salmonella enterica from farm animals in England and Wales. AB - This study is focused on real-time detection of gyrA mutations and of the presence of class I integrons in a panel of 100 veterinary isolates of Salmonella enterica from farm animals. The isolates were selected on the basis of resistance to nalidixic acid, representing a variety of the most prevalent serotypes in England and Wales. In addition, organic solvent (cyclohexane) resistance in these isolates was investigated in an attempt to elucidate the presence of efflux pump mechanisms. The most prevalent mutation among the isolates studied was Asp87-Asn (n = 42), followed by Ser83-Phe (n = 38), Ser83-Tyr (n = 12), Asp87-Tyr (n = 4), and Asp87-Gly (n = 3). Two distinct subpopulations were identified, separated at the 1-mg/liter breakpoint for ciprofloxacin: 86% of isolates with mutations in codon 83 showed MICs of >or=1 mg/liter, while 89.8% of isolates with mutations in codon 87 presented MICs of or=2.0 mg/liter. Thirty-four isolates contained class I integrons, with 71% of the S. enterica serovar Typhimurium isolates and 6.9% of isolates belonging to other serotypes containing such elements. The methods used represent sensitive ways of investigating the presence of gyrA mutations and of detecting class-I integrons in Salmonella isolates. The results can be obtained in less than 1 h from single colonies without the need for purifying DNA. PMID- 11923379 TI - One-point population analysis and effect of osmolarity on detection of hetero vancomycin-resistant Staphylococcus aureus. AB - The original screening test and one-point population analysis (OPPA) were compared for the detection of hetero-vancomycin-resistant Staphylococcus aureus, and the influence of osmolarity on those tests was evaluated. The positivity rates and the reproducibilities were similar for both tests. The addition of NaCl increased the MIC as well as the positivity rates for both tests and the reproducibility of OPPA. PMID- 11923380 TI - Aspergillus galactomannan antigen in the cerebrospinal fluid of bone marrow transplant recipients with probable cerebral aspergillosis. AB - The Aspergillus galactomannan test was performed on cerebrospinal fluid and serum samples from 5 patients with probable cerebral aspergillosis and from 16 control patients. Cerebrospinal fluid galactomannan levels were significantly higher in aspergillosis patients, and most galactomannan was produced intrathecally. Comparison of serum galactomannan values in pulmonary and cerebral aspergillosis patients showed significant overlapping. Detection of Aspergillus galactomannan in cerebrospinal fluid may be diagnostic of cerebral aspergillosis. PMID- 11923381 TI - Performance of four chromogenic urine culture media after one or two days of incubation compared with reference media. AB - Four chromogenic urine culture media were compared to culture on blood agar, MacConkey agar, and CLED (cysteine-, lactose-, and electrolyte-deficient) agar for detection of uropathogens in 1,200 urine specimens. After 2 nights of incubation, 96% of all isolates were recovered on blood agar, 96% were recovered on CLED agar, 92% were recovered on CPS ID2, 96% were recovered on CHROMagar Orientation from BBL, 95% were recovered on CHROMagar Orientation from The CHROMagar Company, and 95% were recovered on Chromogenic UTI Medium. PMID- 11923382 TI - Novel IS6110 insertion sites in the direct repeat locus of Mycobacterium tuberculosis clinical strains from the St. Petersburg area of Russia and evolutionary and epidemiological considerations. AB - A modification of spoligotyping with primers derived from the direct repeat (DR) and IS6110 sequences was used to identify IS6110 insertions in the DR locus of Mycobacterium tuberculosis clinical strains from the St. Petersburg area of Russia. Novel IS6110 insertions were identified: (i) in two epidemiologically unlinked Beijing family strains, an asymmetrical direct insertion in DR37; (ii) in a non-Beijing strain, an asymmetrical insertion in the opposite orientation in DR38; (iii) in another non-Beijing strain, a direct insertion in DR38 and one in the opposite orientation in DR14 (DR numbering is according to standard spoligotyping). Our results strengthen an observation that the DR locus structure is extremely conserved in the Beijing genotype. Asymmetrical insertions prevented detection of the adjacent spacer by standard spoligotyping. This, therefore, should be taken into consideration when similar spoligoprofiles that differ only in signals 37 and 38 are interpreted. PMID- 11923383 TI - Prospective evaluation of the new chromogenic medium Candida ID, in comparison with Candiselect, for isolation of molds and isolation and presumptive identification of yeast species. AB - We conducted a prospective evaluation of Candida ID chromogenic medium (bioMerieux, Marcy l'Etoile, France) with 786 clinical specimens in comparison with Candiselect medium (Bio-Rad, Marnes la Coquette, France). Candida ID chromogenic medium identified 97.7% of Candida albicans strains; enabled presumptive identification of C. tropicalis, C. lusitaniae, C. guillermondii, and C. kefyr and better detection of yeast combinations (11.4% more often); and was more sensitive for the isolation of filamentous fungi (17.7% more often). However, Candida ID chromogenic medium appeared to be less selective vis-a-vis bacteria, with bacterial colonies sometimes pigmented blue. PMID- 11923384 TI - Molecular evidence of Helicobacter cinaedi organisms in human gastric biopsy specimens. AB - One hundred twenty-six urease-negative gastric biopsy specimens were evaluated for the presence of Helicobacter genus-specific 16S ribosomal DNA (rDNA) and H. pylori-specific glmM DNA sequences by PCR. The species specificity of the glmM PCR assay was demonstrated, as H. pylori was the only Helicobacter species that yielded the expected glmM amplicon. Most urease-negative specimens (118 of 126 specimens) lacked Helicobacter DNA. However, 8 of 126 urease-negative specimens contained Helicobacter 16S rDNA. In order to identify the Helicobacter species present in urease-negative gastric biopsy specimens, 16S rDNA amplicons were cloned and sequenced. Sequence comparisons were performed by analyses of the sequences in public sequence databases. Two samples contained 16S rDNA that was identified as H. cinaedi with 100% identity and that spanned approximately 400 bp (398 and 398 bp, respectively). In contrast, multiple differences (97% identity; 390 of 398 bp) were observed with H. pylori 16S rDNA in this region. This finding was verified by sequencing an overlapping 537-bp fragment within the 5' portion of 16S rDNA. Although the clinical findings were consistent with H. pylori infection (e.g., duodenal ulcer disease), rapid urease testing and DNA sequence analyses suggested the presence of H. cinaedi organisms and the absence of H. pylori in two human antral biopsy specimens. This study represents the first report of an enteric urease-negative helicobacter in the human stomach. Although these organisms were previously associated with extragastric infections, the roles of these organisms in the pathogenesis of chronic gastritis or peptic ulcer disease remain unclear. PMID- 11923386 TI - Evaluation of the Abbott LCx quantitative assay for measurement of human immunodeficiency virus RNA in plasma. AB - Plasma human immunodeficiency virus RNA in 491 clinical specimens was measured by LCx. There was a strong correlation with the results provided by other methods (r(2) values of 0.93 with Cobas-Monitor version 1.5 and of 0.95 with Quantiplex version 3.0). However, values were uniformly higher with LCx than with Quantiplex when non-B subtypes were tested. PMID- 11923385 TI - Evaluation of a triplex PCR assay to discriminate Staphylococcus aureus from coagulase-negative Staphylococci and determine methicillin resistance from blood cultures. AB - A triplex PCR targeting the 16S rRNA, mecA, and nuc genes was developed for identification of staphylococci and detection of methicillin resistance. After validation of the assay with a collection of strains of staphylococci and enterococci (n = 169), the assay was evaluated with cultures of blood with gram positive cocci from 40 patients. Accurate results were obtained for 59 (98%) of 61 cultures within 6 h of growth detection. PMID- 11923387 TI - Introduction of new clones of penicillin-nonsusceptible Streptococcus pneumoniae in Hong Kong. AB - Analysis of penicillin-nonsusceptible Streptococcus pneumoniae (PNSP) isolates in Hong Kong by use of a combination of antibiogram typing, serotyping, multilocus sequence typing, and pulsed-field gel electrophoresis indicated that the dissemination of PNSP was the result of the spread of international clones: variants of the Spain(23F)-1 or Spain(6B)-2 clones were the predominant PNSP isolates from 1994 to 1997 and remained so, but Taiwan(19F)-14 and Taiwan serotype 6B clones were disseminated in Hong Kong in 1999 and 2000. Concomitant changes in antibiotic susceptibility profiles, with the rate of susceptibility to chloramphenicol rising from 10% in the period from 1994 to 1997 to 31% (P < 0.001) in 1999 and 2000, were noted to accompany the shift of clones. PMID- 11923388 TI - Community-acquired Acinetobacter baumannii bacteremia in adult patients in Taiwan. AB - We determined the clinical and microbiologic characteristics of community acquired Acinetobacter baumannii bacteremia in 19 adult patients. We found that malignancy was the most frequent underlying disease. The overall mortality rate was 58%. All 14 available isolates were identified as genomic species 2 (A. baumannii) by 16S ribosomal DNA sequencing and were found to be genetically distinct by pulsed-field gel electrophoresis. PMID- 11923389 TI - Four strains of Escherichia coli O157:H7 isolated from patients during an outbreak of disease associated with ground beef: importance of evaluating multiple colonies from an outbreak-associated product. AB - This report describes the investigation of a ground-beef-associated outbreak that involved five genetically distinct patient strains of Escherichia coli O157:H7. Human and product isolates were evaluated by pulsed-field gel electrophoresis (PFGE) with two endonucleases. The multiple-strain etiology of this outbreak underscores the importance of isolating and evaluating multiple colonies from outbreak-related products and comparing two endonuclease PFGE patterns of all product and human isolates identified during outbreak periods. This investigation emphasizes the importance of interviewing all confirmed and suspected case patients during the outbreak period, regardless of the PFGE pattern of their isolate, to confirm or rule out an epidemiologic link to the outbreak. PMID- 11923391 TI - Epidemic strains of Shigella sonnei biotype g carrying integrons. AB - Class 2 integrons (Tn7) were found in all randomly selected epidemic (n = 27) and preepidemic (n = 13) strains of multiresistant Shigella sonnei biotype g. A class 1 integron was also found in two epidemic strains. Gene cassettes within these integrons account for resistance to commonly used therapeutic agents. PMID- 11923390 TI - Detection and characterization of rotaviruses in hospitalized neonates in Blantyre, Malawi. AB - In five separate fecal collections spanning three years, group A rotaviruses were detected by enzyme-linked immunosorbent assay in 35 (25%) of 142 specimens obtained from nondiarrheic, hospitalized neonates in Blantyre, Malawi. Molecular characterization of each strain identified, for the first time in neonates, a short electropherotype, genotype P[6], G8 strain type, similar to the dominant, cocirculating community strain detected in symptomatic infants in Blantyre. Partial sequence analysis of the VP4 and NSP4 genes of neonatal and community strains failed to identify changes which could explain the differences in clinical outcome. Neonatal serotype G8 rotaviruses should be considered as potential rotavirus vaccine candidates for use in Malawi. PMID- 11923392 TI - Use of phylogenetic analysis of hepatitis C virus (HCV) hypervariable region 1 sequences to trace an outbreak of HCV in an autodialysis unit. AB - Hemodialysis patients are at high risk of infection by hepatitis C virus (HCV). The aim of this study was to investigate an HCV outbreak that occurred in an autodialysis unit by using epidemiological and molecular methods. Seroconversion to HCV antibody (anti-HCV) was observed in two patients over an 18-month period; two other patients had previously been recorded as anti-HCV positive. All four patients involved in the outbreak were tested for HCV RNA, and hepatitis C genotype determination was accomplished by a reverse hybridization assay. Furthermore, part of hypervariable region 1 (HVR1) of the hepatitis C genome was amplified and sequenced in samples from all HCV RNA-positive patients. Phylogenetic analysis of the nucleotide sequences obtained was carried out in order to investigate any possible epidemiological linkages among patients. The nucleotide sequences of the HVR1 regions of both newly infected patients were found to be identical to sequences of samples from previously recorded anti-HCV positive original patients, suggesting that they were infected by the same isolate. Molecular and epidemiological analysis suggested that nosocomial patient to-patient transmission was the most likely explanation for the virus spread in the autodialysis unit under study. PMID- 11923393 TI - Production of BRO beta-lactamases and resistance to complement in European Moraxella catarrhalis isolates. AB - Of the 419 Moraxella catarrhalis isolates collected during the 1997-1999 European SENTRY surveillance study, 385 (92%) were beta-lactamase positive. Twenty-two (5.7%) produced BRO-2 beta-lactamase. Twenty-one new mutations were found in the putative promoter region of the bro genes. Nineteen percent of all isolates tested were complement sensitive. Resistance to beta-lactams is not linked to the phylogenetic lineages associated with susceptibility to complement. PMID- 11923395 TI - Rapid particle agglutination test for human immunodeficiency virus: hospital based evaluation. AB - The performance of a rapid particle agglutination test for human immunodeficiency virus (HIV) (Capillus HIV type 1 [HIV-1]/HIV-2) on hospital samples is compared with enzyme-linked immunosorbent assays. The test had a sensitivity and specificity of 99 and 98.9%, respectively. In addition, the test was reactive on plasma samples from all individuals infected with HIV-1 subtype C. This test can safely be used for voluntary counseling and testing in India. PMID- 11923394 TI - Emerging extended-spectrum beta-lactamases in Proteus mirabilis. AB - Beta-lactamase production was detected in 147 (52%) of 282 consecutive nonduplicate Proteus mirabilis isolates obtained over a 1-year period from the S. Matteo Hospital of Pavia (northern Italy). Seventy isolates (48% of the beta lactamase producers) were found to produce extended-spectrum beta-lactamases (ESBLs), identified as PER-1 (first report in this species) and TEM-52 in 52 and 18 isolates, respectively. Analysis of clonal diversity of the ESBL producers suggested different spreading patterns for the two ESBL determinants. PMID- 11923396 TI - Probable mother-to-infant transmission of Pneumocystis carinii f. sp. hominis infection. AB - A mother and her 4.5-week-old infant had Pneumocystis carinii pneumonia contemporaneously. Genotyping of P. carinii f. sp. hominis DNA at three independent loci showed the same genotype in samples from mother and infant. These data suggest transmission of P. carinii organisms from the mother to her infant. PMID- 11923397 TI - Equine abortion associated with the Borrelia parkeri-B. turicatae tick-borne relapsing fever spirochete group. AB - Direct amplification and sequencing of the 16S rRNA gene and a variable region of the flagellin gene from fetal liver-associated spirochetes belonging to the Borrelia parkeri-B. turicatae tick-borne relapsing fever spirochete group with a late-term abortion in a mare are described. PMID- 11923398 TI - Serratia ficaria endophthalmitis. AB - We report a case of Serratia ficaria endophthalmitis in a 73-year-old man. The patient's ocular history included a chemical burn, glaucoma, and corneal transplantation. S. ficaria is part of the fig tree ecosystem and is rarely isolated from clinical specimens. When it has been previously implicated as an agent of disease, the patients have been treated successfully and there have been no complications. In our patient, however, the infection resulted in the loss of the infected eye. This case illustrates that S. ficaria infection in a compromised patient can have serious consequences. PMID- 11923399 TI - Prevalence of the Sat gene among clinical isolates of Shigella spp. causing travelers' diarrhea: geographical and specific differences. PMID- 11923400 TI - Anticoagulants interfere with PCR used to diagnose invasive aspergillosis. PMID- 11923401 TI - Detection of Staphylococcus aureus Clinical Isolates Harboring the ica Gene Cluster Needed for Biofilm Establishment. PMID- 11923402 TI - Improving real-time PCR genotyping assays by asymmetric amplification. PMID- 11923403 TI - Detection of staphylococci with reduced susceptibility to glycopeptides. PMID- 11923404 TI - Complete deletion of the neurotrophin receptor p75NTR leads to long-lasting increases in the number of basal forebrain cholinergic neurons. AB - Cholinergic neurons innervating cortical structures are among the most affected neuronal populations in Alzheimer's disease. In rodents, they express high levels of the neurotrophin receptor p75NTR. We have analyzed cholinergic septohippocampal neurons of the medial septal nucleus in p75exonIII (partial p75NTR knock-out) and p75exonIV (complete p75NTR knock-out) mice, in their original genetic background and in congenic strains. At postnatal day 15, the p75exonIII mutation leads to a moderate increase (+13%) in these neurons among littermates only after back-crossing in a C57BL/6 background. In contrast, the null p75exonIV mutation, which prevents expression of both the full-length and the shorter p75NTR isoforms, results in a 28% neuronal increase, independent of genetic background. The incomplete nature of the p75NTR mutation used previously, coupled with difficulties in delineating the mouse medial septum and the impact of the genetic background on cell numbers, all contribute to explain previous difficulties in establishing the role of p75NTR in regulating cholinergic neuron numbers in the mouse forebrain. PMID- 11923405 TI - Rapid neuromodulatory actions of integrin ligands. AB - Extracellular matrix (ECM) proteins and their receptors, the integrins, actively participate in the control of many fundamental cellular functions in the developing nervous system, including the regulation of cell migration, differentiation, and survival and the control of neurite outgrowth. ECM-integrin interactions in the mature nervous system are commonly considered to be more static in nature and of little importance in the regulation of neuronal function. In contrast, we demonstrate that integrins and their ligands are capable of rapid neuromodulatory actions. Specifically, we show that integrin ligands can alter neuronal pacemaker properties, intracellular free Ca2+ levels, and voltage-gated Ca2+ currents in a matter of minutes. These findings indicate that ECM-integrin interactions play a dynamic role in regulating the physiological status of mature neurons, a process that may contribute to synaptic plasticity, neural regeneration, and neuropathology. PMID- 11923406 TI - Alterations in exocytosis induced by neuronal Ca2+ sensor-1 in bovine chromaffin cells. AB - A variety of Ca2+ binding proteins are known to play an integral role in catecholamine release from synapses as well as secretory cells, such as chromaffin cells. The Drosophila protein frequenin and its mammalian homolog neuronal Ca2+ sensor-1 (NCS-1) belong to a family of Ca2+ sensors with EF hands that bind Ca2+ and then interact with other proteins. Frequenin/NCS-1 has been shown to enhance exocytotic activity in addition to altering Ca2+ channel regulation. To better understand how NCS-1 regulates stimulus-secretion coupling, bovine chromaffin cells were infected with Semliki Forest virus (SFV) vectors containing the rat NCS-1 gene. Cells were studied in the perforated whole-cell patch-clamp configuration. Membrane capacitance was monitored as an indicator of exocytosis-endocytosis. Exocytosis elicited by membrane depolarization was not significantly different between cells infected with SFV expressing green fluorescent protein (GFP) or GFP plus NCS-1, except that the overexpression of NCS-1 resulted in a faster rundown in exocytosis. When cells were stimulated with histamine, NCS-1 overexpression led to higher exocytosis, as well as [Ca2+]i elevation. Immunocytochemistry showed a similar distribution of NCS-1 and phosphatidylinositol 4-kinase beta (PI4Kbeta). NCS-1 and PI4Kbeta coimmunoprecipitate, opening up the possibility that the two proteins directly interact. These results suggest that NCS-1 may regulate cellular activity through the modulation of the phosphatidylinositol signaling pathway. PMID- 11923407 TI - Noradrenergic depletion potentiates beta -amyloid-induced cortical inflammation: implications for Alzheimer's disease. AB - Degeneration of locus ceruleus (LC) neurons and reduced levels of norepinephrine (NE) in LC projection areas are well known features of Alzheimer's disease (AD); however, the consequences of those losses are not clear. Because inflammatory mediators contribute to AD pathogenesis and because NE can suppress inflammatory gene expression, we tested whether LC loss influenced the brain inflammatory gene expression elicited by amyloid beta (Abeta). Adult rats were injected with the selective neurotoxin N-(2-chloroethyl)-N-ethyl-2 bromobenzylamine (DSP4) to induce LC death and subsequently injected in the cortex with Abeta (aggregated 1 42 peptide). DSP4 treatment potentiated the Abeta-dependent induction of inflammatory nitric oxide synthase (iNOS), interleukin (IL)-1beta, and IL-6 expression compared with control animals. In contrast, the induction of cyclooxygenase-2 expression was not modified by DSP4 treatment. In control animals, injection of Abeta induced iNOS primarily in microglial cells, whereas in DSP4-treated animals, iNOS was localized to neurons, as is observed in AD brains. Injection of Abeta increased IL-1beta expression initially in microglia and at later times in astrocytes, and expression levels were greater in DSP4 treated animals than in controls. The potentiating effects of DSP4 treatment on iNOS and IL-1beta expression were attenuated by coinjection with NE or the beta adrenergic receptor agonist isoproterenol. These data demonstrate that LC loss and NE depletion augment inflammatory responses to Abeta and suggest that LC loss in AD is permissive for increased inflammation and neuronal cell death. PMID- 11923408 TI - Synaptically released acetylcholine evokes Ca2+ elevations in astrocytes in hippocampal slices. AB - Recent results have demonstrated the existence of bidirectional communication between glial cells and neurons. We investigated in brain slices whether rat hippocampal astrocytes respond to acetylcholine synaptically released by an extrinsic pathway. We stimulated the stratum oriens/alveus, which contains cholinergic afferents from the septum and diagonal band of Broca, and recorded whole-cell membrane currents and intracellular Ca2+ levels of astrocytes located in the hippocampal stratum oriens. Nerve-fiber stimulation evoked a long-lasting inward current and increased the Ca2+ levels in astrocytes. Both astrocytic responses were abolished by tetrodotoxin or Cd2+ and were increased by 4 aminopyridine, indicating that the responses were attributable to synaptically released neurotransmitter. The inward current was inhibited by glutamate transporter antagonists, indicating that it was attributable to the electrogenic glutamate transporter activity. The synaptically evoked intracellular Ca2+ elevations were not affected by glutamate receptor antagonists but were abolished by atropine, indicating that they were mediated by muscarinic cholinergic receptors. Thapsigargin prevented the Ca2+ elevation but did not modify the inward current, indicating that the Ca2+ signal was attributable to intracellular Ca2+ mobilization. These results indicate that hippocampal astrocytes respond to acetylcholine released by synaptic terminals. The synaptically released acetylcholine acts on muscarinic receptors, mobilizing Ca2+ from the intracellular stores. Different regions in the recorded astrocytes showed independent stimulus-induced Ca2+ variations, suggesting the existence of subcellular domains in the astrocytic responses evoked by the synaptic cholinergic activity. Therefore, our results show the existence of cholinergic neuron-astrocyte signaling and suggest that astrocytes are a target of axonal inputs from different brain areas. PMID- 11923409 TI - The age-related decrease in CNS remyelination efficiency is attributable to an impairment of both oligodendrocyte progenitor recruitment and differentiation. AB - The age-associated decrease in the efficiency of CNS remyelination has clear implications for recovery from demyelinating diseases such as multiple sclerosis (MS) that may last for several decades. Developing strategies to reverse the age associated decline requires the identification of how the regenerative process is impaired. We addressed whether remyelination becomes slower because of an impairment of recruitment of oligodendrocyte progenitors (OPs) or, as is the case in some MS lesions, an impairment of OP differentiation into remyelinating oligodendrocytes. The OP response during remyelination of focal, toxin-induced CNS demyelination in young and old rats was compared by in situ hybridization using probes to two OP-expressed mRNA species: platelet-derived growth factor alpha receptor and the OP transcription factor myelin transcription factor 1 (MyT1). We found that the expression patterns for the two OP markers are very similar and reveal a delay in the colonization of the demyelinated focus with OPs in the old animals compared with the young animals. By comparing the mRNA expression pattern of MyT1 with that of the myelin proteins myelin basic protein and Gtx, we found that in the old animals there is also a delay in OP differentiation that increases with longer survival times. These results indicate that the age-associated decrease in remyelination efficiency occurs because of an impairment of OP recruitment and the subsequent differentiation of the OPs into remyelinating oligodendrocytes, and that strategies aimed at ameliorating the age associated decline in remyelination efficiency will therefore need to promote both components of the regenerative process. PMID- 11923410 TI - G-protein alpha subunit isoforms couple differentially to receptors that mediate presynaptic inhibition at rat hippocampal synapses. AB - Presynaptic receptors that are coupled to heterotrimeric G-proteins are found throughout the brain and are responsible for modulating synaptic transmission. At least 10 G-protein-coupled receptors (GPCRs) reduce transmission in hippocampal neurons. Additionally, hippocampal neurons express up to 17 different Galpha, Gbeta, and Ggamma subunits, making for a striking array of possible heterotrimer compositions and GPCR-heterotrimer interactions. The identity of the Galpha subunit is likely a critical determinant in coupling specificity between GPCRs and their molecular effectors mediating presynaptic inhibition. We studied the role of four Galpha(i/o) subunits (Galpha(o1), Galpha(i1,) Galpha(i2), and Galpha(i3)) in mediating presynaptic inhibition in hippocampal neurons by expressing pertussis toxin-insensitive (PTx-ins) Galpha(i/o) mutants. PTx treatment of these cells disrupts coupling of endogenous subunits, leaving only the mutant Galpha subunits to couple with native GPCRs and betagamma subunits. Successful rescue of presynaptic inhibition indicates that the expressed mutant Galpha subunit can couple to the GPCR of interest. All four PTx-ins Galpha subunits rescued presynaptic inhibition by adenosine A1 receptors. A PTx-ins Galpha subunit also rescued adenosine A1-mediated inhibition of spontaneous vesicle fusion frequency. Of the remaining GPCRs tested, cannabinoid CB1, somatostatin, and GABA(B) receptors displayed an alpha subunit-dependent selectivity in binding to G-protein heterotrimers, whereas group III metabotropic glutamate receptor-mediated inhibition was not rescued by expression of any of the four PTx-ins Galpha subunits. Differential coupling of G-protein alpha subunits may be a means of achieving specificity between different GPCRs and their molecular targets for mediating presynaptic inhibition. PMID- 11923411 TI - Specificity of glomerular targeting by olfactory sensory axons. AB - Axons from olfactory sensory neurons (OSNs) expressing a specific odorant receptor (OR) project to specific subsets of glomeruli in the olfactory bulb (for review, see Mombaerts, 1999, 2001). The aim of this study was to examine the trajectories that subsets of axons from OSNs expressing the same OR follow within the olfactory nerve and olfactory nerve layer (ONL) of adult mice. Using confocal microscopy, we generated serial reconstructions of axons from M72-IRES-tauGFP expressing OSNs as they coursed within the ONL and into glomeruli. GFP-expressing axons were loosely aggregated in the outer ONL; however, as they entered the inner ONL, the majority fasciculated with other GFP-expressing axons before entering the glomerular neuropil. Although the vast majority of axons entered the glomerulus from the directly apposed ONL, some followed tortuous courses through and/or around adjacent glomeruli before terminating in the target glomerulus. Similar observations were made on subpopulations of axons in M71-IRES-tauGFP and P2-IRES-tauGFP mice. Ultrastructural analyses of labeled M72 glomeruli showed no evidence of axodendritic synapses other than those with GFP-labeled axon terminals. These data are consistent with the notion that OSN axons are highly precise in targeting glomeruli and that glomeruli, in turn, are highly homogeneous with regard to the OR expressed by the innervating OSNs. Because some single axons could follow idiosyncratic trajectories to the target glomerulus, it appears that stable homotypic fasciculation is not a prerequisite for correct targeting. PMID- 11923412 TI - The toll-like receptor TLR4 is necessary for lipopolysaccharide-induced oligodendrocyte injury in the CNS. AB - The immediate or innate immune response is the first line of defense against diverse microbial pathogens and requires the expression of recently discovered toll-like receptors (TLRs). TLR4 serves as a specific receptor for lipopolysaccharide (LPS) and is localized on the surface of a subset of mammalian cells. Although innate immunity is a necessary host defense against microbial pathogens, the consequences of its activation in the CNS can be deleterious, as we show here in a developing neural model. We examined the major non-neuronal cell types in the CNS for expression of TLR4 and found that microglia expressed high levels, whereas astrocytes and oligodendrocytes expressed none. Consistent with TLR4 expression solely in microglia, we show that microglia are the only CNS glial cells that bind fluorescently tagged lipopolysaccharide. Lipopolysaccharide led to extensive oligodendrocyte death in culture only under conditions in which microglia were present. To determine whether TLR4 is necessary for lipopolysaccharide-induced oligodendrocyte death in mixed glial cultures, we studied cultures generated from mice bearing a loss-of-function mutation in the tlr4 gene. Lipopolysaccharide failed to induce oligodendrocyte death in such cultures, in contrast to the death induced in cultures from wild-type mice. Finally, stereotactic intracerebral injection of lipopolysaccharide into the developing pericallosal white matter of immature rodents resulted in loss of oligodendrocytes and hypomyelination and periventricular cysts. Our data provide a general mechanistic link between (1) lipopolysaccharide and similar microbial molecular motifs and (2) injury to oligodendrocytes and myelin as occurs in periventricular leukomalacia and multiple sclerosis. PMID- 11923414 TI - The Ras1-mitogen-activated protein kinase signal transduction pathway regulates synaptic plasticity through fasciclin II-mediated cell adhesion. AB - Ras proteins are small GTPases with well known functions in cell proliferation and differentiation. In these processes, they play key roles as molecular switches that can trigger distinct signal transduction pathways, such as the mitogen-activated protein kinase (MAPK) pathway, the phosphoinositide-3 kinase pathway, and the Ral-guanine nucleotide dissociation stimulator pathway. Several studies have implicated Ras proteins in the development and function of synapses, but the molecular mechanisms for this regulation are poorly understood. Here, we demonstrate that the Ras-MAPK pathway is involved in synaptic plasticity at the Drosophila larval neuromuscular junction. Both Ras1 and MAPK are expressed at the neuromuscular junction, and modification of their activity levels results in an altered number of synaptic boutons. Gain- or loss-of-function mutations in Ras1 and MAPK reveal that regulation of synapse structure by this signal transduction pathway is dependent on fasciclin II localization at synaptic boutons. These results provide evidence for a Ras-dependent signaling cascade that regulates fasciclin II-mediated cell adhesion at synaptic terminals during synapse growth. PMID- 11923413 TI - Myosin-1c interacts with hair-cell receptors through its calmodulin-binding IQ domains. AB - Myosin-1c plays an essential role in adaptation of hair-cell mechanoelectrical transduction. To mediate adaptation, myosin-1c must interact directly or indirectly with other components of the transduction apparatus, including the mechanically gated transduction channel. As a first step toward identifying myosin-1c receptors, we used recombinant myosin-1c fragments to identify specific binding sites in hair cells and to biochemically characterize their interaction with myosin-1c. Myosin-1c fragments bound to tips of hair-cell stereocilia, the location of transduction and adaptation. Surprisingly, this interaction did not depend on the C-terminal tail of myosin-1c, proposed previously to be the receptor-binding site of the molecule. Instead, the interaction of myosin-1c with stereociliary receptors depended on its calmodulin-binding IQ domains. This interaction was blocked by calmodulin, which probably bound to a previously unoccupied IQ domain of myosin-1c. The calcium-sensitive binding of calmodulin to myosin-1c may therefore modulate the interaction of the adaptation motor with other components of the transduction apparatus. PMID- 11923415 TI - Disease-specific human glycine receptor alpha1 subunit causes hyperekplexia phenotype and impaired glycine- and GABA(A)-receptor transmission in transgenic mice. AB - Hereditary hyperekplexia is caused by disinhibition of motoneurons resulting from mutations in the ionotropic receptor for the inhibitory neurotransmitter glycine (GlyR). To study the pathomechanisms involved in vivo, we generated and analyzed transgenic mice expressing the hyperekplexia-specific dominant mutant human GlyR alpha1 subunit 271Q. Tg271Q transgenic mice, in contrast to transgenic animals expressing a wild-type human alpha1 subunit (tg271R), display a dramatic phenotype similar to spontaneous and engineered mouse mutations expressing reduced levels of GlyR. Electrophysiological analysis in the ventral horn of the spinal cord of tg271Q mice revealed a diminished GlyR transmission. Intriguingly, an even larger reduction was found for GABA(A)-receptor-mediated inhibitory transmission, indicating that the expression of this disease gene not only affects the glycinergic system but also leads to a drastic downregulation of the entire postsynaptic inhibition. Therefore, the transgenic mice generated here provide a new animal model of systemic receptor interaction to study inherited and acquired neuromotor deficiencies at different functional levels and to develop novel therapeutic concepts for these diseases. PMID- 11923416 TI - Cell type- and input-specific differences in the number and subtypes of synaptic GABA(A) receptors in the hippocampus. AB - Networks of parvalbumin (PV)-expressing basket cells are implicated in synchronizing cortical neurons at various frequencies, through GABA(A) receptor mediated synaptic action. These cells are interconnected by GABAergic synapses and gap junctions, and converge with a different class of cholecystokinin expressing, PV-negative basket cells onto pyramidal cells. To define the molecular specializations in the synapses of the two basket cell populations, we used quantitative electron microscopic immunogold localization of GABA(A) receptors. Synapses formed by PV-positive basket cells on the somata of pyramidal cells had several-fold higher density of alpha1 subunit-containing receptors than synapses made by PV-negative basket cells, most of which were immunonegative. The density of the beta2/3 subunits was similar in the two populations of synapse, indicating similar overall receptor density. Synapses interconnecting parvalbumin expressing basket cells contained a 3.6 times higher overall density of GABA(A) receptor (beta2/3 subunits) and 3.2 times higher density of alpha1 subunit labeling compared with synapses formed by boutons of PV-positive basket cells on pyramidal cells. Thus, PV-positive basket cells mainly act through alpha1 subunit containing GABA(A) receptors, but the receptor density depends on the postsynaptic cell type. These observations, together with previously reported enrichment of the alpha2 subunit-containing receptors in synapses made by PV negative basket cells, indicate that the number and subtypes of GABA(A) receptors present in different synapse populations are regulated by both presynaptic and postsynaptic influences. The high number of GABA(A) receptors in synapses on basket cells might contribute to the precisely timed phasing of basket cell activity. PMID- 11923417 TI - Involvement of the alpha3 subunit in central nicotinic binding populations. AB - The alpha3 subunit gene was one of the first neuronal nicotinic acetylcholine receptor (nAChR) subunits to be cloned (Boulter et al., 1986), but direct evidence of alpha3 subunit contributions to mammalian central nAChR populations has not been presented. The studies reported here used mice engineered to contain a null mutation in the alpha3 nAChR subunit gene (Xu et al., 1999) to examine the involvement of the alpha3 subunit in central nAChR populations. Heterologously expressed alpha3beta2 and alpha3beta4 nAChRs are pharmacologically similar to native [125I]alpha-conotoxin MII (alpha-CtxMII)-binding and 3-(2(S) azetidinylmethoxy)pyridine dihydrochloride (A85380)-resistant [125I]epibatidine binding nAChR subtypes, respectively. The hypothesis that both native sites are alpha3-subtype nAChRs was tested using quantitative autoradiography in alpha3 null mutant mice. Somewhat surprisingly, deletion of the alpha3 nAChR subunit gene did not affect expression of the great majority of [125I]alpha-CtxMII binding sites, indicating that they do not correspond to heterologously expressed alpha3beta2 nAChRs. The only exception to this was observed in the habenulointerpeduncular tract, where alpha3-dependent [125I]alpha-CtxMII binding was observed. This finding may suggest the presence of an additional, minor nicotinic population in this pathway. In contrast, most -resistant [125I]epibatidine-binding nAChRs were dependent on alpha3 gene expression, suggesting that they do indeed correspond to an alpha3 nAChR subtype. However, widespread but lower levels of alpha3-independent -resistant [125I]epibatidine binding were also seen. Again, this may indicate the existence of an additional, minor population of non-alpha3 -resistant sites. PMID- 11923418 TI - Regulated cationic channel function in Xenopus oocytes expressing Drosophila big brain. AB - Big brain (bib) is a neurogenic gene that when mutated causes defects in cell fate determination during Drosophila neurogenesis through an unknown mechanism. The protein Big Brain (BIB) has sequence identity with the major intrinsic protein family that includes the water- and ion-conducting aquaporin channels. We show here that BIB expressed heterologously in Xenopus oocytes provides a voltage insensitive, nonselective cation channel function with permeability to K+ > Na+ >> tetraethylammonium. The conductance, activated in response to endogenous signaling pathways in BIB-expressing oocytes, is decreased after treatment with 20 microm insulin and is enhanced with 10 microm lavendustin A, a tyrosine kinase inhibitor. Western blot analysis confirms that BIB is tyrosine-phosphorylated. Both tyrosine phosphorylation and the potentiating effect of lavendustin A are removed by partial deletion of the C terminus (amino acids 317-700). Current activation is not observed in control oocytes or in oocytes expressing a nonfunctional mutant (BIB E71N) that appears to be expressed on the plasma membrane by confocal microscopy and Western blotting. These results indicate that BIB can participate in tyrosine kinase-regulated transmembrane signaling and may suggest a role for membrane depolarization in the neurogenic function of BIB in early development. PMID- 11923419 TI - Nicotinic acetylcholine receptors interact with dopamine in induction of striatal long-term depression. AB - The dorsal striatum participates in motor function and stimulus-response or "habit" learning. Acetylcholine (ACh) is a prominent neurotransmitter in the striatum and exerts part of its actions through nicotinic cholinergic receptors. Activation of these receptors has been associated with the enhancement of learning and certainly is instrumental in habitual use of nicotine. Nicotinic receptors have also been suggested to be a possible therapeutic target for disorders of the basal ganglia. In this report we show that the activation of nicotinic acetylcholine receptors in the dorsal striatum contributes to dopamine (DA)- and activity-dependent changes in synaptic efficacy. High-frequency activation of glutamatergic synapses onto striatal neurons results in a long-term depression (LTD) of synaptic efficacy that is dependent on the activation of dopamine receptors. This stimulation also produces robust increases in extracellular dopamine concentration as well as strong activation of cholinergic striatal interneurons. Antagonists of nicotinic acetylcholine receptors inhibit striatal LTD. However, on coapplication of dopamine reuptake inhibitors with nicotinic receptor antagonists, activity-induced striatal LTD is restored. Dopamine release is modulated by activation of nicotinic receptors in the dorsal striatum, and activation of nicotinic receptors during high-frequency synaptic activation appears to be capable of interacting with dopaminergic actions that lead to striatal LTD. Our results suggest that stimulation of mechanisms involved in striatal synaptic plasticity is an important role for striatal nicotinic acetylcholine receptors and that these mechanisms may contribute to the enhancement of learning and habit formation produced by nicotine intake. PMID- 11923420 TI - Glutamate but not glycine agonist affinity for NMDA receptors is influenced by small cations. AB - NMDA receptor currents desensitize in an agonist-dependent manner when either the glutamate or glycine agonist is subsaturating. This may result from a conformational change in the NMDA receptor protein that lowers glutamate and glycine binding site affinity induced by co-agonist binding, channel opening, or ion permeation. We have used whole-cell voltage clamp of cultured hippocampal neurons with agonist paired-pulse protocols to demonstrate that glutamate and glycine dissociate 7.9- and 6.8-fold slower in the absence of their respective co agonists than when their co-agonists are present. Paired-pulse and desensitization protocols were used to show that co-agonist binding and channel opening are sufficient to cause a reduction in glycine affinity, but extracellular sodium or magnesium binding was required in addition to conformational changes leading to channel opening to reduce glutamate binding site affinity. Use of cesium or potassium as the major extracellular cation prevented the reduction of glutamate affinity. In addition, the use of choline-, sodium-, or cesium-based intracellular solutions did not alter desensitization characteristics, indicating that the site responsible for reduction of glutamate affinity is not in the intracellular domain. The fact that the reduction of glutamate affinity is dependent on certain small extracellular cations whereas the reduction of glycine affinity is insensitive to such cations indicates that conformational changes induced by the binding of glutamate are not completely paralleled by the conformational changes induced by glycine. Although glutamate and glycine are essential co-agonists, these data suggest that they have differential roles in the process of NMDA receptor activation. PMID- 11923421 TI - Localization of activator protein-1 complex with DNA binding activity in mitochondria of murine brain after in vivo treatment with kainate. AB - To elucidate mechanisms underlying mitochondrial dysfunctions induced by glutamate, we have examined the effects of in vivo treatment with the ionotropic glutamate receptor agonist kainate on localization of the transcription factor activator protein-1 (AP-1) in mitochondria as well as nuclei of murine brain. A systemic administration of kainate dramatically enhanced AP-1 DNA binding in both mitochondrial and nuclear extracts of mouse cerebral cortex and hippocampus 1 hr to 3 d later. Unlabeled AP-1 probe selectively competed for AP-1 DNA binding in mitochondrial extracts of cortex and hippocampus obtained from mice injected with kainate. Supershift and immunoblotting analyses revealed participation of c-Fos, Fos-B, and Jun-B proteins in potentiation by kainate of mitochondrial AP-1 DNA binding in cortex and hippocampus. An immunohistochemical study demonstrated marked expression by kainate of c-Fos protein in the pyramidal and dentate granular layers, whereas an immunoelectron microscopic analysis showed localization of c-Fos protein within mitochondria, as well as nuclei, of the CA1 pyramidal and dentate granular cells in hippocampus obtained 2 hr after the administration of kainate. Mitochondrial AP-1 DNA binding was inhibited by particular unlabeled oligonucleotides containing sequences similar to the AP-1 site found in the noncoding region of mitochondrial DNA. Kainate markedly potentiated binding of radiolabeled oligonucleotide probes containing sequences effective in competing for AP-1 DNA binding in hippocampal mitochondrial extracts. These results suggest that kainate may facilitate expression of the AP 1 complex and subsequent translocation into mitochondria to participate in mechanisms associated with transcriptional regulation of mitochondrial DNA in murine hippocampus. PMID- 11923422 TI - Failure of brain-derived neurotrophic factor-dependent neuron survival in mouse trisomy 16. AB - The neurotrophin, brain derived neurotrophic factor (BDNF), exerts multiple effects on the development and maintenance of the nervous system, including regulating synaptic plasticity and promoting neuron survival. Here we report the selective failure of BDNF-dependent survival in cultured hippocampal neurons from the trisomy 16 (Ts16) mouse, an animal model of Down syndrome. This failure is accompanied by overexpression of a truncated, kinase-deficient isoform (T1) of the BDNF receptor tyrosine receptor kinase B (trkB). Adenovirus-mediated introduction of exogenous full-length trkB into Ts16 neurons fully restored BDNF dependent survival, whereas exogenous truncated trkB expression in normal, euploid neurons reproduced the Ts16 BDNF signaling failure. Thus, the failure of Ts16 neurons to respond to BDNF is caused by dysregulation of trkB isoform expression. Such a neurotrophin signaling defect could contribute to developmental and degenerative disorders of the nervous system. PMID- 11923423 TI - A proximal promoter domain containing a homeodomain-binding core motif interacts with multiple transcription factors, including HoxA5 and Phox2 proteins, and critically regulates cell type-specific transcription of the human norepinephrine transporter gene. AB - Expression of the norepinephrine transporter (NET), which mediates the reuptake of norepinephrine into presynaptic nerve terminals, is restricted to noradrenergic (NA) neurons. We have demonstrated previously that the 9.0 kb upstream sequences and the first intron residing in the 5' untranslated area are critical for high-level and NA cell-specific transcription. Here, using transient transfection assays, we show that 4.0 kb of the 5' upstream sequences contains sufficient genetic information to drive reporter gene expression in an NA cell type-specific manner. Three functional domains appear to be potentially important for the regulation of human NET (hNET) gene transcription: an upstream enhancer region at -4.0 to -3.1 kb, a proximal domain at -133 to -75 bp, and a middle silencer region between these two domains. DNase I footprinting analysis of the proximal promoter region shows that a subdomain at -128 to -80 bp is protected in a cell-specific manner. We provide evidence that multiple protein factors interact with the proximal promoter domain to critically regulate the transcriptional activity of the hNET gene. In the middle of this proximal subdomain resides a homeodomain (HD)-binding core motif, which interacts with HD factors, including Phox2a and HoxA5, in an NA-specific manner. Cotransfection analyses suggest that HoxA5 and Phox2a may transactivate the hNET gene promoter. Together with previous studies indicating direct activation of dopamine beta hydroxylase transcription by Phox2a/2b, the present results support a model whereby Phox2 proteins may coordinately regulate the phenotypic specification of NA neurons by activating both NA biosynthetic and reuptake genes. PMID- 11923425 TI - Descending 5-hydroxytryptamine raphe inputs repress the expression of serotonergic neurons and slow the maturation of inhibitory systems in mouse embryonic spinal cord. AB - Spontaneous synchronous rhythmic activities are a common feature of immature neuronal networks. Although the mechanisms underlying such activities have been studied extensively, whether they might be controlled by modulatory information remains questionable. Here, we investigated the role of descending serotonergic (5-HT) inputs from the medulla to the spinal cord in the maturation of rhythmic activity. We found that in spinal cords maintained, as a whole, in organotypic culture without the medulla, the maturation of spontaneous activity is similar to that found in spinal cords developed in utero. Interestingly, in organotypic cultures without the medulla (i.e., devoid of descending inputs), numerous intraspinal neurons expressed 5-HT, unlike in spinal cords cultivated in the presence of the medulla or matured in utero. We demonstrated that this 5-HT expression was specifically dependent on the absence of 5-HT fibers and was repressed by 5-HT itself via activation of 5-HT(1A) receptors. Finally, to verify whether the expression of 5-HT intraspinal neurons could compensate for the lack of descending 5-HT fibers and play a role in the development of spontaneous activity, we blocked the 5-HT synthesis using p-chlorophenylalanine methyl ester in cultures devoid of the medulla. Surprisingly, we found that this pharmacological treatment did not prevent the development of spontaneous activity but accelerated the maturation of intraspinal inhibition at the studied stages. Together, our data indicate that descending 5-HT raphe inputs (1) repress the expression of spinal serotonergic neurons and (2) slow the maturation of inhibitory systems in mouse spinal cord. PMID- 11923424 TI - Cdk5/p35 regulates neurotransmitter release through phosphorylation and downregulation of P/Q-type voltage-dependent calcium channel activity. AB - Cyclin-dependent kinase 5 (Cdk5) is a proline-directed serine/threonine kinase with close structural homology to the mitotic Cdks. The complex of Cdk5 and p35, the neuron-specific regulatory subunit of Cdk5, plays important roles in brain development, such as neuronal migration and neurite outgrowth. Moreover, Cdk5 is thought to be involved in the promotion of neurodegeneration in Alzheimer's disease. Cdk5 is abundant in mature neurons; however, its physiological functions in the adult brain are unknown. Here we show that Cdk5/p35 regulates neurotransmitter release in the presynaptic terminal. Both Cdk5 and p35 were abundant in the synaptosomes. Roscovitine, a specific inhibitor of Cdk5 in neurons, induced neurotransmitter release from the synaptosomes in response to membrane depolarization and enhanced the EPSP slopes in rat hippocampal slices. The electrophysiological study using each specific inhibitor of the voltage dependent calcium channels (VDCCs) and calcium imaging revealed that roscovitine enhanced Ca2+ influx from the P/Q-type VDCC. Moreover, Cdk5/p25 phosphorylated the intracellular loop connecting domains II and III (L(II-III)) between amino acid residues 724 and 981 of isoforms cloned from rat brain of the alpha1A subunit of P/Q-type Ca2+ channels. The phosphorylation inhibited the interaction of L(II-III) with SNAP-25 and synaptotagmin I, which were plasma membrane soluble N-ethylmaleimide-sensitive factor attachment protein (SNAP) receptor (SNARE) proteins and were required for efficient neurotransmitter release. These results strongly suggest that Cdk5/p35 inhibits neurotransmitter release through the phosphorylation of P/Q-type VDCC and downregulation of the channel activity. PMID- 11923426 TI - Cell density and N-cadherin interactions regulate cell proliferation in the sensory epithelia of the inner ear. AB - Sensory hair cells in the inner ears of nonmammalian vertebrates can regenerate after injury. In many species, replacement hair cells are produced by the proliferation of epithelial supporting cells. Thus, the ability of supporting cells to undergo renewed proliferation is a key determinant of regenerative ability. The present study used cultures of isolated inner ear sensory epithelia to identify cellular signals that regulate supporting cell proliferation. Small pieces of sensory epithelia from the chicken utricle were cultured in glass microwells. Under those conditions, cell proliferation was inversely related to local cell density. The signaling molecules N-cadherin, beta-catenin, and focal adhesion kinase were immunolocalized in the cultured epithelial cells, and high levels of phosphotyrosine immunoreactivity were present at cell-cell junctions and focal contacts of proliferating cells. Binding of microbeads coated with a function-blocking antibody to N-cadherin inhibited ongoing proliferation. The growth of epithelial cells was also affected by the density of extracellular matrix molecules. The results suggest that cell density, cell-cell contact, and the composition of the extracellular matrix may be critical influences on the regulation of sensory regeneration in the inner ear. PMID- 11923427 TI - Rapid, experience-dependent changes in levels of synaptic zinc in primary somatosensory cortex of the adult mouse. AB - Electrophysiological studies have established that the adult cerebral cortex undergoes immediate functional reorganizations after perturbations of the sensory periphery. These activity-dependent modifications are thought to be mediated via the rapid regulation of the synaptic strength of existing connections. Recent studies have implicated synaptic zinc as contributing to activity-dependent mechanisms of cortical plasticity, such as long-term potentiation and long-term depression, by virtue of its potent ability to modulate glutamatergic neurotransmission. To investigate the role of synaptic zinc in cortical plasticity, we examined changes in the barrel-specific distribution of zinc in axon terminals innervating the primary somatosensory cortex of adult mice at different time points after whisker plucking. In layer IV of normal adult mice, zinc staining in the barrel field was characterized by intense staining in inter barrel septae and low levels of staining in barrel hollows. Within 3 hr, and up to 1 week after the removal of a row of whiskers, zinc staining increased significantly in barrel hollows corresponding to the plucked whiskers. With longer survival times, levels of zinc staining gradually declined in deprived barrel hollows, returning to normal levels by 2-3 weeks after whisker removal. Increased levels of zinc staining in deprived barrel hollows were highly, negatively correlated with the length of whiskers as they regrew. These results indicate that levels of synaptic zinc in the neocortex are rapidly regulated by changes in sensory experience and suggest that zinc may participate in the plastic changes that normally occur in the cortex on a moment-to-moment basis. PMID- 11923428 TI - Spontaneous retinal activity is tonic and does not drive tectal activity during activity-dependent refinement in regeneration. AB - During development, waves of activity periodically spread across retina to produce correlated activity that is thought to drive activity-dependent ordering in optic fibers. We asked whether similar waves of activity are produced in the retina of adult goldfish during activity-dependent refinement by regenerating optic fibers. Dual-electrode recordings of spontaneous activity were made at different distances across retina but revealed no evidence of retinal waves in normal retina or during regeneration. Retinal activity was tonic and lacked the episodic bursting associated with waves. Cross-correlation analysis showed that the correlated activity that was normally restricted to near neighbors (typically seen across 100-200 microm and absent at >500 microm) was not altered during regeneration. The only change associated with regeneration was a twofold reduction in ganglion cell firing rates. Because spontaneous retinal activity is known to be sufficient to generate refinement during regeneration in goldfish, we examined its effect on tectal activity. In normal fish, acutely eliminating retinal activity with TTX rapidly reduced tectal unit activity by >90%. Surprisingly, during refinement at 4-6 weeks, eliminating retinal activity had no detectable effect on tectal activity. Similar results were obtained in recordings from torus longitudinalis. After refinement at 3 months, tectal activity was again highly dependent on ongoing retinal activity. We conclude that spontaneous retinal activity drives tectal cells in normal fish and after regeneration but not during activity-dependent refinement. The implications of these results for the role of presynaptic activity in refinement are considered. PMID- 11923429 TI - Caspase inhibitors attenuate 1-methyl-4-phenylpyridinium toxicity in primary cultures of mesencephalic dopaminergic neurons. AB - Parkinson's disease is characterized by a loss of dopaminergic nigrostriatal neurons. This neuronal loss is mimicked by the neurotoxin 1-methyl-4 phenylpyridinium (MPP+). MPP+ toxicity is mediated through inhibition of mitochondrial complex I, decreasing ATP production, and upregulation of oxygen radicals. There is evidence that the cell death induced by MPP+ is apoptotic and that inhibition of caspases may be neuroprotective. In primary cultures of rat mesencephalic dopaminergic neurons, MPP+ treatment decreased the number of surviving dopaminergic neurons in the cultures and the ability of the neurons to take up [3H]dopamine ([3H]DA). Caspase inhibition using the broad-spectrum inhibitor benzyloxycarbonyl-Val-Ala-Asp-fluoromethylketone (zVAD-fmk) spared MPP+ treated dopaminergic neurons and increased somatic size. There was a partial restoration of neurite length in zVAD-fmk-treated cultures, but little restoration of [3H]DA uptake. Peptide inhibitors of caspases 2, 3, and 9, but not of caspase 1, caused significant neuroprotection. Two novel caspase inhibitors were tested for neuroprotection, a broad spectrum inhibitor and a selective caspase 3 inhibitor; both inhibitors increased survival to >90% of control. No neuroprotection was observed with an inactive control compound. MPP+ treatment caused chromatin condensation in dopaminergic neurons and increased expression of activated caspase 3. Inhibition of caspases with either zVAD-fmk or a selective caspase 3 inhibitor decreased the number of apoptotic profiles, but not expression of the active caspase. We conclude that MPP+ toxicity in primary dopaminergic neurons involves activation of a pathway terminating in caspase 3 activation, but that other mechanisms may underlie the neurite loss. PMID- 11923430 TI - Estrogen regulates the development of brain-derived neurotrophic factor mRNA and protein in the rat hippocampus. AB - During development, estrogen has a variety of effects on morphological and electrophysiological properties of hippocampal neurons. Brain-derived neurotrophic factor (BDNF) also plays an important role in the survival and differentiation of neurons during development. We examined the effects of gonadectomy with and without estrogen replacement on the mRNA and protein of BDNF and its receptor, trkB, during early postnatal development of the rat hippocampus. We used immunocytochemistry to demonstrate that estrogen receptor alpha (ERalpha) and BDNF were localized to the same cells within the developing hippocampus. BDNF and ERalpha were colocalized in pyramidal cells of the CA3 subregion and to a lesser extent in CA1. To determine whether BDNF mRNA was regulated by estrogen during development, we gonadectomized male rat pups at postnatal day 0 (P0) and examined mRNA and protein levels from P0 to P25 using real-time reverse transcription-PCR and Western blot analysis. After gonadectomy, BDNF mRNA levels are significantly reduced on P7, but after treatment of gonadectomized animals with estradiol benzoate on P0, levels at all ages were similar to those in intact animals. BDNF mRNA changes after gonadectomy are accompanied by an increase in the levels of BDNF protein, which were reduced by estrogen treatment at P0. We also examined the effect of postnatal estrogen treatment on trkB. There were no significant changes in trkB mRNA or protein in gonadectomized or estrogen-replaced animals. These results suggest that a direct interaction may exist between ERalpha and BDNF to alter hippocampal physiology during development in the rat. PMID- 11923431 TI - Metabotropic glutamate receptor 5 upregulation in A-fibers after spinal nerve injury: 2-methyl-6-(phenylethynyl)-pyridine (MPEP) reverses the induced thermal hyperalgesia. AB - Metabotropic glutamate receptor 5 (mGluR5) protein increased after sciatic nerve section in ipsilateral L4 and L5 DRG neuronal profiles, with most of the increase occurring in myelinated A-fiber somata. mGluR5 also increased in lamina II of the ipsilateral spinal cord and the proximal sciatic nerve stump in this model. After L5 spinal nerve ligation, mGluR5 immunoreactivity increased dramatically not only in damaged L5 but also in the neighboring undamaged L4. Interestingly, after partial sciatic nerve section, mGluR5 expression did not change in either L4 or L5 DRG neuronal profiles. Both spinal nerve ligation and sciatic nerve partial section produced significant mechanical and thermal hyperalgesia and tactile allodynia. After partial sciatic nerve section, the mGluR5-specific antagonist 2 methyl-6-(phenylethynyl)-pyridine (MPEP) had no effect on any of these behaviors. However, after L5 spinal nerve ligation, although MPEP failed to alter the induced tactile allodynia or mechanical hyperalgesia, it dose dependently reversed the developed thermal hyperalgesia. Therefore, reversal of thermal hyperalgesia by MPEP correlates with increased mGluR5 in lumbar DRG A-fiber somata after nerve injury. Furthermore, A-fibers in the uninjured L4 DRG after L5 spinal nerve ligation that have increased mGluR5 are the same A-fibers that newly express vanilloid receptor 1 after such injury. Together, these results suggest that, after L5 spinal nerve injury, mGluR5 expression on A-fibers is essential to the development of thermal hyperalgesia. After partial nerve section, however, it is unlikely that thermal responses are mediated through mGluR5 because no such increase in mGluR5 is detected in this model and MPEP is ineffective. PMID- 11923432 TI - Target interaction regulates distribution and stability of specific mRNAs. AB - Several factors regulate export of mRNAs from neuronal cell bodies. Using in situ hybridization and RT-PCR, we examined how target interaction influences the distribution of mRNAs expressed in sensory neurons (SNs) of Aplysia maintained in cell culture. Interaction with a synaptic target has two effects on the distribution of mRNA encoding an SN-specific peptide, sensorin: the target affects the accumulation of sensorin mRNA at the axon hillock and the stability of sensorin mRNA exported to distal sites. Synapse formation with motor neuron L7 results in the accumulation of high levels of sensorin mRNA in the axon hillock of the SN and in SN neurites contacting L7. SNs cultured alone or in contact with motor neuron L11, with which no synapses form, show a more uniform distribution of sensorin mRNA in the cytoplasm of the SN cell body, with little expression in neurites. Contact with L7 or L11 had little or no effect on the distribution of two other mRNAs in the cytoplasm of SN cell bodies. Sensorin mRNA exported to SN neurites after 1 d in culture is more stable when the SN contacts L7 compared with SN neurites that contact L11. After removal of the SN cell body, the amounts of sensorin mRNA already exported to the neurites are greater when neurites contact L7 compared with neurites in contact with L11. The results indicate that target interaction and synapse formation regulate both the accumulations of specific mRNAs destined for export and their stability at distant sites. PMID- 11923433 TI - Enriched odor exposure increases the number of newborn neurons in the adult olfactory bulb and improves odor memory. AB - In the mammalian forebrain, most neurons originate from proliferating cells in the ventricular zone lining the lateral ventricles, including a discrete area of the subventricular zone (SVZ). In this region, neurogenesis continues into adulthood. Most of the cells generated in the SVZ are neuronal precursors with progeny that migrate rostrally along a pathway known as the rostral migratory stream before they reach the main olfactory bulb (MOB) where they differentiate into local interneurons. The olfactory system thus provides an attractive model to investigate neuronal production and survival, processes involving interplay between genetic and epigenetic influences. The present study was conducted to investigate whether exposure to an odor-enriched environment affects neurogenesis and learning in adult mice. Animals housed in either a standard or an odor enriched environment for 40 d were injected intraperitoneally with bromodeoxyuridine (BrdU) to detect proliferation among progenitor cells and to follow their survival in the MOB. The number of BrdU-labeled neurons was not altered 4 hr after a single BrdU injection. In contrast, the number of surviving progenitors 3 weeks after BrdU injection was markedly increased in animals housed in an enriched environment. This effect was specific because enriched odor exposure did not influence hippocampal neurogenesis. Finally, we showed that adult mice housed in odor-enriched cages display improved olfactory memory without a change in spatial learning performance. By maintaining a constitutive turnover of granule cells subjected to modulation by environmental cues, ongoing bulbar neurogenesis could be associated with improved olfactory memory. PMID- 11923434 TI - Pathological CNS autoimmune disease triggered by traumatic spinal cord injury: implications for autoimmune vaccine therapy. AB - Lymphocytes respond to myelin proteins after spinal cord injury (SCI) and may contribute to post-traumatic secondary degeneration. However, there is increasing evidence that autoreactive T-lymphocytes may also convey neuroprotection and promote functional recovery after CNS injury. To clarify the role of myelin autoreactive lymphocytes after SCI, we performed contusion injuries in the thoracic spinal cord of transgenic (Tg) mice in which >95% of all CD4+ T lymphocytes are reactive with myelin basic protein (MBP). We observed significantly impaired recovery of locomotor and reflex function in Tg mice compared with non-Tg (nTg) littermates. Measures of functional impairment in Tg mice correlated with significantly less white matter at the injury site, and morphometric comparisons of injured Tg and nTg spinal cords revealed increased rostrocaudal lesion expansion (i.e., secondary degeneration) in Tg mice. Rostrocaudal to the impact site in SCI-nTg mice, demyelination was restricted to the dorsal funiculus, i.e., axons undergoing Wallerian degeneration. The remaining white matter appeared normal. In contrast, lymphocytes were colocalized with regions of demyelination and axon loss throughout the white matter of SCI-Tg mice. Impaired neurological function and exacerbated neuropathology in SCI-Tg mice were associated with increased intraspinal production of proinflammatory cytokine mRNA; neurotrophin mRNA was not elevated. These data suggest that endogenous MBP-reactive lymphocytes, activated by traumatic SCI, can contribute to tissue injury and impair functional recovery. Any neuroprotection afforded by myelin-reactive T-cells is likely to be an indirect effect mediated by other non CNS-reactive lymphocytes. Similar to the Tg mice in this study, a subset of humans that are genetically predisposed to autoimmune diseases of the CNS may be adversely affected by vaccine therapies designed to boost autoreactive lymphocyte responses after CNS trauma. Consequently, the safe implementation of such therapies requires that future studies define the mechanisms that control T-cell function within the injured CNS. PMID- 11923435 TI - Intravitreal injection of the attenuated pseudorabies virus PRV Bartha results in infection of the hamster suprachiasmatic nucleus only by retrograde transsynaptic transport via autonomic circuits. AB - Intravitreal injection of the attenuated strain of pseudorabies virus (PRV Bartha) results in transneuronal spread of virus to a restricted set of central nuclei in the rat and mouse. We examined the pattern of central infection in the golden hamster after intravitreal inoculation with a recombinant strain of PRV Bartha constructed to express enhanced green fluorescent protein (PRV 152). Neurons in a subset of retinorecipient nuclei [i.e., suprachiasmatic nucleus (SCN), intergeniculate leaflet, olivary pretectal nucleus (OPN), and lateral terminal nucleus] and autonomic nuclei [i.e., paraventricular hypothalamic nucleus and Edinger-Westphal nucleus (EW)] are labeled by late stages of infection. Infection of the EW precedes infection in retinorecipient structures, raising the possibility that the SCN becomes infected by retrograde transsynaptic infection via autonomic (i.e., EW) circuits. We tested this hypothesis in two ways: (1) by removing the infected eye 24 hr after PRV 152 inoculation, well before viral infection first appears in the SCN; and (2) by examining central infection after intravitreal PRV 152 injection in animals with ablation of the EW. The pattern and time course of central infection were unchanged after enucleation, whereas EW ablation before intravitreal inoculation eliminated viral infection in the SCN. The results of EW lesions along with known connections between EW, OPN, and SCN indicate that intravitreal injection of PRV Bartha produces a retrograde infection of the autonomic innervation of the eye, which subsequently labels a restricted set of retinorecipient nuclei via retrograde trans-synaptic infection. These results, taken together with other genetic data, indicate that the mutations in PRV Bartha render the virus incapable of anterograde transport. PRV Bartha is thus a retrograde transsynaptic marker in the CNS. PMID- 11923436 TI - Neural correlates of encoding space from route and survey perspectives. AB - The neural mechanisms underlying ground-level spatial navigation have been investigated, but little is known about other kinds of spatial navigation. Functional magnetic resonance imaging was used to identify differences in brain activation for two types of spatial information, information from the ground level perspective (route) and information from a global perspective (survey). Participants were scanned during the encoding of two different virtual reality environments, one from each perspective. Comparisons of brain activation during route and survey encoding suggested that both types of information recruited a common network of brain areas, but with important differences. Survey encoding recruited a subset of areas recruited by route encoding, but with greater activation in some areas, including inferior temporal cortex and posterior superior parietal cortex. Route encoding, in contrast, recruited regions that were not activated by survey encoding, including medial temporal lobe structures, anterior superior parietal cortex, and postcentral gyrus. These differences in brain activation are associated with differences in memory performance for the two types of spatial information and contribute to specification of brain components of spatial knowledge. PMID- 11923437 TI - Gene expression profiling reveals alterations of specific metabolic pathways in schizophrenia. AB - Dysfunction of the dorsal prefrontal cortex (PFC) in schizophrenia may be associated with alterations in the regulation of brain metabolism. To determine whether abnormal expression of genes encoding proteins involved in cellular metabolism contributes to this dysfunction, we used cDNA microarrays to perform gene expression profiling of all major metabolic pathways in postmortem samples of PFC area 9 from 10 subjects with schizophrenia and 10 matched control subjects. Genes comprising 71 metabolic pathways were assessed in each pair, and only five pathways showed consistent changes (decreases) in subjects with schizophrenia. Reductions in expression were identified for genes involved in the regulation of ornithine and polyamine metabolism, the mitochondrial malate shuttle system, the transcarboxylic acid cycle, aspartate and alanine metabolism, and ubiquitin metabolism. Interestingly, although most of the metabolic genes that were consistently decreased across subjects with schizophrenia were not similarly decreased in haloperidol-treated monkeys, the transcript encoding the cytosolic form of malate dehydrogenase displayed prominent drug-associated increases in expression compared with untreated animals. These molecular analyses implicate a highly specific pattern of metabolic alterations in the PFC of subjects with schizophrenia and raise the possibility that antipsychotic medications may exert a therapeutic effect, in part, by normalizing some of these changes. PMID- 11923438 TI - The neural correlates of moral sensitivity: a functional magnetic resonance imaging investigation of basic and moral emotions. AB - Humans are endowed with a natural sense of fairness that permeates social perceptions and interactions. This moral stance is so ubiquitous that we may not notice it as a fundamental component of daily decision making and in the workings of many legal, political, and social systems. Emotion plays a pivotal role in moral experience by assigning human values to events, objects, and actions. Although the brain correlates of basic emotions have been explored, the neural organization of "moral emotions" in the human brain remains poorly understood. Using functional magnetic resonance imaging and a passive visual task, we show that both basic and moral emotions activate the amygdala, thalamus, and upper midbrain. The orbital and medial prefrontal cortex and the superior temporal sulcus are also recruited by viewing scenes evocative of moral emotions. Our results indicate that the orbital and medial sectors of the prefrontal cortex and the superior temporal sulcus region, which are critical regions for social behavior and perception, play a central role in moral appraisals. We suggest that the automatic tagging of ordinary social events with moral values may be an important mechanism for implicit social behaviors in humans. PMID- 11923439 TI - Functional asymmetries in ON and OFF ganglion cells of primate retina. AB - Functional asymmetries in the ON and OFF pathways of the primate visual system were examined using simultaneous multi-electrode recordings from dozens of retinal ganglion cells (RGCs) in vitro. Light responses of RGCs were characterized using white noise stimulation. Two distinct functional types of cells frequently encountered, one ON and one OFF, had non-opponent spectral sensitivity, relatively high response gain, transient light responses, and large receptive fields (RFs) that tiled the region of retina recorded, suggesting that they belonged to the same morphological cell class, most likely parasol. Three principal functional asymmetries were observed. (1) Receptive fields of ON cells were 20% larger in diameter than those of OFF cells, resulting in higher full field sensitivity. (2) ON cells had faster response kinetics than OFF cells, with a 10-20% shorter time to peak, trough and zero crossing in the biphasic temporal impulse response. (3) ON cells had more nearly linear light responses and were capable of signaling decrements, whereas OFF cells had more strongly rectifying responses that provided little information about increments. These findings suggest specific mechanistic asymmetries in retinal ON and OFF circuits and differences in visual performance on the basis of the activity of ON and OFF parasol cells. PMID- 11923440 TI - Imaging attentional modulation of pain in the periaqueductal gray in humans. AB - Pain is an unpleasant sensory and emotional experience usually triggered by stimulation of peripheral nerves and often associated with actual or potential tissue damage. It is well known that pain perception for patients and normal subjects can be modulated by psychological factors, such as attention, stress, and arousal. Our understanding of how this modulation occurs at a neuroanatomical level is poor. Here we neuroanatomically defined a key area in the network of brain regions active in response to pain that is modulated by attention to the painful stimulus. High-resolution functional magnetic resonance imaging was used to define brain activation to painful heat stimulation applied to the hand of nine normal subjects within the periaqueductal gray region. Subjects were asked to either focus on or distract themselves from the painful stimuli, which were cued using colored lights. During the distraction condition, subjects rated the pain intensity as significantly lower compared with when they attended to the stimulus. Activation in the periaqueductal gray was significantly increased during the distraction condition, and the total increase in activation was predictive of changes in perceived intensity. This provides direct evidence supporting the notion that the periaqueductal gray is a site for higher cortical control of pain modulation in humans. PMID- 11923441 TI - Impaired conditioned fear and enhanced long-term potentiation in Fmr2 knock-out mice. AB - FRAXE mental retardation results from expansion and methylation of a CCG trinucleotide repeat located in exon 1 of the X-linked FMR2 gene, which results in transcriptional silencing. The product of FMR2 is a member of a family of proteins rich in serine and proline, members of which have been associated with transcriptional activation. We have developed a murine Fmr2 gene knock-out model by replacing a fragment containing parts of exon 1 and intron 1 with the Escherichia coli lacZ gene, placing lacZ under control of the Fmr2 promoter. Expression of lacZ in the knock-out animals indicates that Fmr2 is expressed in several tissues, including brain, bone, cartilage, hair follicles, lung, tongue, tendons, salivary glands, and major blood vessels. In the CNS, Fmr2 expression begins at the time that cells in the neuroepithelium differentiate into neuroblasts. Mice lacking Fmr2 showed a delay-dependent conditioned fear impairment. Long-term potentiation (LTP) was found to be enhanced in hippocampal slices of Fmr2 knock-out compared with wild-type littermates. To our knowledge, this mouse knock-out is the first example of an animal model of human mental retardation with impaired learning and memory performance and increased LTP. Thus, although a number of studies have suggested that diminished LTP is associated with memory impairment, our data suggest that increased LTP may be a mechanism that leads to impaired cognitive processing as well. PMID- 11923442 TI - CNS site of action and brainstem circuitry responsible for the intravenous effects of nicotine on gastric tone. AB - The purposes of our study were to determine (1) the effects of intravenous (i.v.) nicotine on gastric mechanical function of anesthetized rats, (2) the CNS site of action of nicotine to produce these effects, (3) the CNS nicotinic acetylcholine receptor (nAChR) subtype(s) responsible for mediating the i.v. effects of nicotine, and (4) the brainstem neurocircuitry engaged by i.v. nicotine for eliciting its gastric effects. This was accomplished by monitoring intragastric pressure (gastric tone) and contractility of the fundus and antrum while administering five doses of i.v. nicotine and microinjecting nicotine into specific brainstem nuclei. Additionally, c-Fos expression in the brainstem after i.v. nicotine and pharmacological agents were used as tools to identify the CNS site and circuitry and reveal the nAChR subtype(s) mediating the gastric effects of nicotine. Using these experimental approaches, we found the following. (1) When given intravenously in doses of 56.5, 113, 226, 452, and 904 nmol/kg, nicotine elicited only inhibitory effects on gastric mechanical function. The most sensitive area of the stomach to nicotine was the fundus, and this effect was mediated by the vagus nerve at doses of 56.5, 113, and 226 nmol/kg. (2) The CNS site of action and nAChR subtype responsible were glutamatergic vagal afferent nerve terminals in the medial subnucleus of the tractus solitarious (mNTS) and alpha4beta2, respectively. (3) The brainstem neurocircuitry that was involved appeared to consist of a mNTS noradrenergic pathway projecting to the dorsal motor nucleus of the vagus (DMV). This pathway seems to be activated via nitriergic interneurons engaged by vagally released glutamate in the mNTS and results in alpha2 adrenergic receptor-mediated inhibition of DMV neurons projecting to the fundus and controlling gastric tone. PMID- 11923443 TI - Parkinson-like neurodegeneration induced by targeted overexpression of alpha synuclein in the nigrostriatal system. AB - Recombinant adeno-associated viral vectors display efficient tropism for transduction of the dopamine neurons of the substantia nigra. Taking advantage of this unique property of recombinant adeno-associated viral vectors, we expressed wild-type and A53T mutated human alpha-synuclein in the nigrostriatal dopamine neurons of adult rats for up to 6 months. Cellular and axonal pathology, including alpha-synuclein-positive cytoplasmic inclusions and swollen, dystrophic neurites similar to those seen in brains from patients with Parkinson's disease, developed progressively over time. These pathological alterations occurred preferentially in the nigral dopamine neurons and were not observed in other nondopaminergic neurons transduced by the same vectors. The degenerative changes were accompanied by a loss of 30-80% of the nigral dopamine neurons, a 40-50% reduction of striatal dopamine, and tyrosine hydroxylase levels that was fully developed by 8 weeks. Significant motor impairment developed in those animals in which dopamine neuron cell loss exceeded a critical threshold of 50-60%. At 6 months, signs of cell body and axonal pathology had subsided, suggesting that the surviving neurons had recovered from the initial insult, despite the fact that alpha-synuclein expression was maintained at a high level. These results show that nigral dopamine neurons are selectively vulnerable to high levels of either wild-type or mutant alpha-synuclein, pointing to a key role for alpha-synuclein in the pathogenesis of Parkinson's disease. Targeted overexpression of alpha synuclein in the nigrostriatal system may provide a new animal model of Parkinson's disease that reproduces some of the cardinal pathological, neurochemical, and behavioral features of the human disease. PMID- 11923444 TI - NG2 is a major chondroitin sulfate proteoglycan produced after spinal cord injury and is expressed by macrophages and oligodendrocyte progenitors. AB - Several extracellular matrix (ECM) molecules have been identified as potent inhibitors of neurite outgrowth in vitro and are believed to limit axonal growth after CNS injury. Recent studies have shown that different members of the chondroitin sulfate proteoglycan (CSPG) class of putatively inhibitory ECM molecules are expressed after a number of CNS injuries. The purpose of this study was to evaluate the relative amounts of individual CSPGs expressed after spinal cord injury (SCI) and identify their cells of origin. Evaluation of total soluble CSPGs 2 weeks after dorsal column lesion in the rat demonstrated that NG2 is highly upregulated and is a major CSPG species. Immunocytochemical analysis further demonstrated that NG2 expression is upregulated within 24 hr of injury, peaks at 1 week, and remains elevated for at least an additional 7 weeks. NG2 expression results from a multicellular response to injury, including both reactive macrophages and oligodendrocyte progenitors; astrocytes were not identified as a major source of NG2. Immunocytochemical analysis of other CSPG family members 7 d after injury showed moderate upregulation of versican, brevican, and neurocan, and downregulation of phosphacan. Axonal tracing experiments demonstrated dense NG2 labeling adjacent to the forward processes of transected corticospinal tract axons in a spatial profile that could restrict axonal growth. Thus, NG2 is a major component of this putatively inhibitory class of ECM molecules expressed at sites of SCI and may restrict axonal regeneration. PMID- 11923445 TI - Electrotonically mediated oscillatory patterns in neuronal ensembles: an in vitro voltage-dependent dye-imaging study in the inferior olive. AB - Spatiotemporal profiles of ensemble subthreshold neuronal oscillation were studied in brainstem slices using high-speed voltage-sensitive dye imaging. After local electrical stimuli, the overall voltage profile demonstrated coherent oscillatory waves that spread over the inferior olive (IO). These oscillations were also observed in concurrently obtained intracellular recordings from IO neurons. Over the first few seconds after the stimuli, the optically recorded oscillations clustered into coherent groups comprising hundreds of neurons. Statistical analysis of the spatial profiles of these clusters revealed size fluctuation around stable core regions that were surrounded by a rim the diameter of which varied in time during the oscillation period. The neuronal ensemble oscillations were calcium derived and had an average frequency range of 1-7 Hz. This rhythmic response demonstrated a different spatiotemporal distribution in the presence of picrotoxin, which induced the merging of neuronal clusters into larger areas of coherent activity. The possibility that such clustering is a consequence of intrinsic oscillations in ensembles of coupled neurons was tested using mathematical modeling. PMID- 11923446 TI - Long-term consequences of switching handedness: a positron emission tomography study on handwriting in "converted" left-handers. AB - Until some decades ago, left-handed children who attended German schools were forced to learn to write with their right hand. To explore the long-term consequences of switching handedness, we studied the functional neuroanatomy of handwriting in 11 adult "converted" left-handers and 11 age-matched right handers. All participants had used exclusively their right hand for writing since early childhood. Using [15O]H2O positron emission tomography, changes in normalized regional cerebral blood flow (rCBF) were assessed while participants repetitively wrote a stereotyped word with their right hand. The kinematics of handwriting did not differ between converted left-handers and right-handers. In innate right-handers, handwriting caused a preponderant left-hemispheric activation of parietal and premotor association areas. In contrast, converted left-handers demonstrated a more bilateral activation pattern with distinct activation foci in the right lateral premotor, parietal, and temporal cortex. Moreover, foci in the right rostral supplementary motor area and the right inferior parietal lobule demonstrated a positive linear relationship between the degree of "left-handedness" and normalized rCBF during right-hand writing. Functional activity in the primary sensorimotor cortex was not affected by handedness. Our findings provide evidence for persisting differences in the functional neuroanatomy of handwriting between right-handers and converted left handers, despite decades of right-hand writing. Right-hemispheric activation in converted left-handers may reflect suppression of unwanted left-hand movements. Alternatively, this activity may represent persistent left-handedness and, as such, demonstrate a hemispheric asymmetry of hand movement representations in cortical motor association areas in relation to the direction and degree of handedness. PMID- 11923447 TI - Bimodal interactions in the superior colliculus of the behaving cat. AB - Bimodal enhancement, a form of nonlinear summation of physiological responses from two sensory modalities, has been demonstrated in the intermediate layers of the superior colliculus (SCi) and is thought to be a manifestation of a neural mechanism underlying behavioral facilitation to such stimuli. Most physiological studies, however, have been performed in anesthetized animals. We tested for bimodal enhancement in the SCi of behaving cats trained to orient to acoustic, visual, and bimodal stimuli. Surprisingly, we never observed the large enhanced responses reported in anesthetized animals, even when we varied the time between presentation of the visual and acoustic stimuli and/or decreased the level of the stimuli. Using three different behavioral paradigms, we found no support for enhanced interactions between auditory and visual modalities. Prominent depressive effects were seen, however, particularly when the cats were required to fixate a visual target during presentation of an acoustic stimulus. PMID- 11923448 TI - Effects of reversible inactivation of the neonatal ventral hippocampus on behavior in the adult rat. AB - Rats with neonatal excitotoxic damage of the ventral hippocampus display in adulthood a variety of abnormalities reminiscent of schizophrenia and are used as an animal model of this disorder. In the present study, we hypothesized that transient inactivation of ventral hippocampal activity during a critical developmental period may be sufficient to disrupt normal maturation of relevant brain systems and produce similar lasting behavioral changes. We infused tetrodotoxin (TTX) or artificial CSF into the ventral hippocampus on postnatal day 7 (P7) and assessed behavioral changes in response to stress, amphetamine, and (+)-5-methyl-10,11-dihydro-5H-dibenzo [a,d] cyclohepten-5,10-imine maleate in juvenile (P35) and young adult (P56) rats. In adulthood, rats infused neonatally with TTX displayed motor hyperactivity after pharmacological stimulation and after stress compared with sham controls. Analogous TTX infusions in adult animals did not alter these behaviors later in life. These data suggest that transient loss of ventral hippocampal function during a critical time in maturation of intracortical connections permanently changes the development of neural circuits mediating certain dopamine- and NMDA-related behaviors. These results represent a potential new model of aspects of schizophrenia without involving a gross anatomic lesion. PMID- 11923449 TI - The physiological role of 5-HT2A receptors in working memory. AB - Dorsolateral prefrontal cortex has an essential role in the cognitive process of working memory, dysfunction of which is considered to be a core deficit in schizophrenia. Although this cortical region is densely innervated with 5-HT2A receptors to which atypical antipsychotic drugs bind with high affinity, little is known of the influence of this serotonin receptor subtype on prefrontal function. We addressed this issue by examining the effects of iontophoresis of selective receptor ligands on prefrontal neurons possessing spatially tuned delay activity, or "memory fields," in monkeys performing a delayed-response task. Memory fields of putative pyramidal cells were attenuated by iontophoresis of 5 HT2A antagonists, which primarily produced a reduction in delay activity for preferred target locations. Conversely, 5-HT2A stimulation by alpha-methyl-5-HT or 5-HT itself, accentuated the spatial tuning of these neurons by producing a modest increase in activity for preferred target locations and/or a reduction in activity for nonpreferred locations. The agonist effects could be reversed by the selective antagonist MDL100,907, and were dose-dependent, such that high levels attenuated spatial tuning by profoundly reducing delay activity. A role for feedforward inhibitory circuitry in these effects was supported by the finding that 5-HT2A blockade also attenuated the memory fields of putative interneurons. We conclude that prefrontal 5-HT2A receptors have a hitherto unrecognized role in the cognitive function of working memory, which involves actions at both excitatory and inhibitory elements within local circuitry. PMID- 11923450 TI - Synchronized neuronal discharge in the basal ganglia of parkinsonian patients is limited to oscillatory activity. AB - It has been proposed that an increase in synchronization between neurons in the basal ganglia contributes to the clinical features of Parkinson's disease (PD). To examine this hypothesis, we looked for correlations in the discharge activity of pairs of neurons in the globus pallidus internus (GPi), globus pallidus externus (GPe), and the substantia nigra pars reticulata (SNr). Recordings were performed in PD patients undergoing functional stereotactic mapping for pallidotomy (eight patients) or subthalamic nucleus deep brain stimulation (four patients). A double-microelectrode setup was used to simultaneously record from neurons separated by distances as small as 250 microm. In the five pallidotomy patients without limb tremor during the procedure, none of the 73 GPi pairs and 15 GPe pairs displayed synchronous activity. In the three pallidotomy patients with limb tremor, 6 of 21 GPi pairs and 5 of 29 GPe pairs displayed oscillatory synchronization in the frequency range of the ongoing limb tremor (3-6 Hz) or at higher frequencies (15-30 Hz). Synchronized activity was not observed in the SNr (10 pairs). The findings indicate that oscillatory synchronization between pairs of GPi or GPe neurons is found in patients with limb tremor. These results also suggest that overt neuronal synchronization, which may be attributable to an increase in direct synaptic connections or common collateral afferent inputs, is not present in the basal ganglia of patients with PD. PMID- 11923451 TI - Direct and indirect excitation of laterodorsal tegmental neurons by Hypocretin/Orexin peptides: implications for wakefulness and narcolepsy. AB - Compelling evidence links the recently discovered hypothalamic peptides Hypocretin/Orexin (Hcrt/Orx) to rapid eye movement sleep (REM) control and the sleep disorder narcolepsy, yet how they influence sleep-related systems is not well understood. We investigated the action of Hcrt/Orx on mesopontine cholinergic (MPCh) neurons of the laterodorsal tegmental nucleus (LDT), a target group whose function is altered in canine narcolepsy and appears pivotal for normal REM and wakefulness. Extracellular recordings from mouse brainstem slices revealed that Hcrt/Orx evoked prolonged firing of LDT neurons. Whole-cell recordings revealed that Hcrt/Orx had actions on both presynaptic neurons and at postsynaptic sites. Hcrt/Orx produced an increase in frequency and amplitude of spontaneous EPSCs without equivalent effect on IPSCs, by triggering action potentials and enhancing spike-evoked synaptic transmission in glutamatergic afferents. Postsynaptically, Hcrt/Orx produced an inward current and an increase in membrane current noise, which were accompanied by a conductance increase. These persisted in TTX, ionotropic glutamate receptor antagonists, and low extracellular calcium. Both presynaptic and postsynaptic actions were specific because they were not mimicked by an Hcrt/Orx fragment, and both actions were observed for cholinergic and noncholinergic LDT neurons. Finally, extracellular recordings during postsynaptic potential blockade demonstrated that postsynaptic actions of Hcrt/Orx alone could evoke prolonged firing. In the context of other recent work, our findings suggest that Hcrt/Orx neurons may coordinate the activity of the entire reticular activating system during waking. Moreover, these findings address specific hypotheses regarding the cellular mechanisms underlying REM disregulation in narcolepsy. PMID- 11923452 TI - Alpha1b-adrenergic receptors control locomotor and rewarding effects of psychostimulants and opiates. AB - Drugs of abuse, such as psychostimulants and opiates, are generally considered as exerting their locomotor and rewarding effects through an increased dopaminergic transmission in the nucleus accumbens. Noradrenergic transmission may also be implicated because most psychostimulants increase norepinephrine (NE) release, and numerous studies have indicated interactions between noradrenergic and dopaminergic neurons through alpha1-adrenergic receptors. However, analysis of the effects of psychostimulants after either destruction of noradrenergic neurons or pharmacological blockade of alpha1-adrenergic receptors led to conflicting results. Here we show that the locomotor hyperactivities induced by d-amphetamine (1-3 mg/kg), cocaine (5-20 mg/kg), or morphine (5-10 mg/kg) in mice lacking the alpha1b subtype of adrenergic receptors were dramatically decreased when compared with wild-type littermates. Moreover, behavioral sensitizations induced by d amphetamine (1-2 mg/kg), cocaine (5-15 mg/kg), or morphine (7.5 mg/kg) were also decreased in knock-out mice when compared with wild-type. Ruling out a neurological deficit in knock-out mice, both strains reacted similarly to novelty, to intraperitoneal saline, or to the administration of scopolamine (1 mg/kg), an anti-muscarinic agent. Finally, rewarding properties could not be observed in knock-out mice in an oral preference test (cocaine and morphine) and conditioned place preference (morphine) paradigm. Because catecholamine tissue levels, autoradiography of D1 and D2 dopaminergic receptors, and of dopamine reuptake sites and locomotor response to a D1 agonist showed that basal dopaminergic transmission was similar in knock-out and wild-type mice, our data indicate a critical role of alpha1b-adrenergic receptors and noradrenergic transmission in the vulnerability to addiction. PMID- 11923453 TI - Cortical synaptic arrangements of the third visual pathway in three primate species: Macaca mulatta, Saimiri sciureus, and Aotus trivirgatus. AB - The koniocellular (K) pathway is one of three pathways from the lateral geniculate nucleus (LGN) to primate visual cortex (V1). K pathway projections to the cytochrome oxidase (CO) blobs of V1 suggest involvement in chromatic processing given reports that the CO blobs in diurnal primates contain cells selective for color. K LGN layers and CO blobs, however, are also well developed in nocturnal primates such as owl monkeys, which are likely to be color blind. Thus, the K pathway plays either different roles in different species or some as yet unidentified common role(s). Because synaptic arrangements underlie functional mechanisms, the purpose of this investigation was to compare the synaptic circuitry related to the K pathway within the CO blobs of two diurnal primates (macaque monkeys and squirrel monkeys) and one nocturnal primate (owl monkey). Presynaptic K axons were labeled with wheat germ agglutinin-HRP, and presynaptic and postsynaptic profiles in CO blobs were identified with post embedding immunocytochemistry for GABA and glutamate. In all three species, K axon terminals are glutamatergic and larger than local axon terminals, suggesting that they have a greater impact on postsynaptic CO blob targets than signals arriving via layer IV from the P or M pathways. A greater proportion of K axons, however, synapse with larger glutamatergic shafts in the diurnal monkeys than in the nocturnal owl monkey, perhaps reflecting the importance of color within the K pathway of these diurnal species. Alternatively, the loss of color vision in the owl monkey could impact K pathway circuitry earlier in the pathway. The basic similarities between K axon circuitry within the CO blobs of the three primate species examined also could indicate that this pathway plays some common role or roles across species. PMID- 11923454 TI - Conservation of spatial memory function in the pallial forebrain of reptiles and ray-finned fishes. AB - The hippocampus of mammals and birds is critical for spatial memory. Neuroanatomical evidence indicates that the medial cortex (MC) of reptiles and the lateral pallium (LP) of ray-finned fishes could be homologous to the hippocampus of mammals and birds. In this work, we studied the effects of lesions to the MC of turtles and to the LP of goldfish in spatial memory. Lesioned animals were trained in place, and cue maze tasks and crucial probe and transfer tests were performed. In experiment 1, MC-lesioned turtles in the place task failed to locate the goal during trials in which new start positions were used, whereas sham animals navigated directly to the goal independently of start location. In contrast, no deficit was observed in cue learning. In experiment 2, LP lesion produced a dramatic impairment in goldfish trained in the place task, whereas medial and dorsal pallium lesions did not decrease accuracy. In addition, none of these pallial lesions produced deficits in cue learning. These results indicate that lesions to the MC of turtles and to the LP of goldfish, like hippocampal lesions in mammals and birds, selectively impair map-like memory representations of the environmental space. Thus, the forebrain structures of reptiles and teleost fish neuroanatomically equivalent to the mammalian and avian hippocampus also share a central role in spatial cognition. Present results suggest that the presence of a hippocampus-dependent spatial memory system is a primitive feature of the vertebrate forebrain that has been conserved through evolution. PMID- 11923455 TI - Natural stimulation of the nonclassical receptive field increases information transmission efficiency in V1. AB - We have investigated how the nonclassical receptive field (nCRF) affects information transmission by V1 neurons during simulated natural vision in awake, behaving macaques. Stimuli were centered over the classical receptive field (CRF) and stimulus size was varied from one to four times the diameter of the CRF. Stimulus movies reproduced the spatial and temporal stimulus dynamics of natural vision while maintaining constant CRF stimulation across all sizes. In individual neurons, stimulation of the nCRF significantly increases the information rate, the information per spike, and the efficiency of information transmission. Furthermore, the population averages of these quantities also increase significantly with nCRF stimulation. These data demonstrate that the nCRF increases the sparseness of the stimulus representation in V1, suggesting that the nCRF tunes V1 neurons to match the highly informative components of the natural world. PMID- 11923456 TI - Cocaine administered into the medial prefrontal cortex reinstates cocaine-seeking behavior by increasing AMPA receptor-mediated glutamate transmission in the nucleus accumbens. AB - One of the major determinants of reinstatement to cocaine use among human addicts is acute reexposure to the drug, which often precipitates cocaine craving and relapse. We used an animal model of cocaine relapse to determine the role of the glutamatergic pathway from the medial prefrontal cortex (mPFC) to the nucleus accumbens in the reinstatement of cocaine-seeking behavior after a cocaine priming injection. Rats were trained to self-administer cocaine intravenously on a second order schedule. Responding was extinguished subsequently by substituting saline for cocaine. During subsequent reinstatement sessions, drug-seeking behavior was assessed after noncontingent priming injections. Results indicated that reinstatement induced by a systemic cocaine injection was blocked by intra mPFC administration of the dopamine antagonist flupenthixol. Consistent with this finding, administration of cocaine directly into the mPFC reinstated cocaine seeking behavior. Administration of cocaine into the nucleus accumbens also reinstated drug seeking, whereas microinjection of cocaine into the neostriatum or lateral septum did not. Reinstatement of cocaine seeking induced by intra-mPFC cocaine was blocked by administration of the AMPA receptor antagonist CNQX into the nucleus accumbens. Administration of the NMDA receptor antagonist AP-5 into the nucleus accumbens had variable effects on reinstatement induced by intra-mPFC cocaine in that AP-5 had no effect in some animals but augmented reinstatement in others. Subsequent experiments showed that intra-accumbal microinjection of AP-5 alone dose-dependently reinstated cocaine seeking. These data indicate that the glutamatergic pathway from the mPFC to the nucleus accumbens plays an important role in cocaine priming-induced reinstatement of drug seeking. Moreover, the present results demonstrate that AMPA and NMDA receptors in the nucleus accumbens have opposing roles in the reinstatement of cocaine-seeking behavior. PMID- 11923458 TI - Cortical activation during spoken-word segmentation in nonreading-impaired and dyslexic adults. AB - We used magnetoencephalography to elucidate the cortical activation associated with the segmentation of spoken words in nonreading-impaired and dyslexic adults. The subjects listened to binaurally presented sentences where the sentence-ending words were either semantically appropriate or inappropriate to the preceding sentence context. Half of the inappropriate final words shared two or three initial phonemes with the highly expected semantically appropriate words. Two temporally and functionally distinct response patterns were detected in the superior temporal lobe. The first response peaked at approximately 100 msec in the supratemporal plane and showed no sensitivity to the semantic appropriateness of the final word. This presemantic N100m response was abnormally strong in the left hemisphere of dyslexic individuals. After the N100m response, the semantically inappropriate sentence-ending words evoked stronger activation than the expected endings in the superior temporal cortex in the vicinity of the auditory cortex. This N400m response was delayed for words starting with the same two or three first few phonemes as the expected words but only until the first evidence of acoustic-phonetic dissimilarity emerged. This subtle delay supports the notion of initial lexical access being based on phonemes or acoustic features. In dyslexic participants, this qualitative aspect of word processing appeared to be normal. However, for all words alike, the ascending slope of the semantic activation in the left hemisphere was delayed by approximately 50 msec as compared with control subjects. The delay in the auditory N400m response in dyslexic subjects is likely to result from presemantic-phonological deficits possibly reflected in the abnormal N100m response. PMID- 11923457 TI - Reduction of stress-induced behavior by antagonism of corticotropin-releasing hormone 2 (CRH2) receptors in lateral septum or CRH1 receptors in amygdala. AB - Although corticotropin-releasing hormone (CRH), a regulator of stress responses, acts through two receptors (CRH1 and CRH2), the role of CRH2 in stress responses remains unclear. Knock-out mice without the CRH2 gene exhibit increased stress like behaviors. This profile could result either directly from the absence of CRH2 receptors or indirectly from developmental adaptations. In the present study, CRH2 receptors were acutely blocked by alpha-helical CRH (alpha(h)CRH, CRH1/CRH2 antagonist; 0, 30, 100, and 300 ng) infusion into the lateral septum (LS), which abundantly expresses CRH2 but not CRH1 receptors. Freezing, locomotor activity, and analgesia were tested after infusion. Intra-LS alpha(h)CRH blocked shock-induced freezing without affecting activity or pain responses; infusions into lateral ventricle or nucleus of the diagonal band had no effects. The same behavioral profile was obtained with d-Phe-CRH((12-41)) (100 ng), another CRH1/CRH2 antagonist. A selective CRH1 antagonist (NBI27914), in doses that reduced freezing on intra-amygdala (central nucleus) infusion (0, 0.2, and 1.0 microg), did not affect freezing when infused into the LS. Ex vivo autoradiography revealed that binding of [125I]sauvagine, a mixed CRH1/CRH2 agonist, was prevented in the LS by previous intra-LS infusion of alpha(h)CRH but not NBI27914. In vitro studies demonstrated that [125I]sauvagine binding in the LS could be inhibited by a CRH1/CRH2 antagonist but not by the selective CRH1 receptor antagonist, confirming that in the LS, alpha(h)CRH antagonized exclusively CRH2 receptors. Acute antagonism of CRH2 receptors in the LS thus produces a behaviorally, anatomically, and pharmacologically specific reduction in stress-induced behavior, in contrast to results of recent knock-out studies, which induced congenital and permanent CRH2 removal. CRH2 receptors may thus represent a potential target for the development of novel CRH system anxiolytics. PMID- 11923459 TI - Development of response timing and direction selectivity in cat visual thalamus and cortex. AB - Single-unit recordings were made in the dorsal lateral geniculate nucleus (LGN) and visual cortex of kittens that were 4-13 weeks of age. Responses to visual stimuli were analyzed to determine the relationship between two related facets of the behaviors of the cells: direction selectivity (DS) and timing. DS depends on timing differences within the receptive field. Cortical DS was present at all ages, but its temporal frequency tuning changed. In kittens, DS was more common at high (approximately 4 Hz) than low ( approximately 1 Hz) temporal frequencies. This is in contrast to adults, in which DS is tuned to low frequencies, more common at 1 Hz than at 4 Hz (Saul and Humphrey, 1992a). In adult cats, the LGN provides the cortex with a wide range of timings that are also observable in cortical receptive fields (Saul and Humphrey, 1990, 1992b; Alonso et al., 2001). In kittens, LGN and cortical timing were immature. Most cells showed long-latency sustained responses. At low temporal frequencies, the variance in timing was small, but at higher frequencies, all timings were well represented. The timing data thus matched the temporal frequency tuning of DS. Kittens show DS at high temporal frequencies because of the abundance of inputs with different timing at high frequencies. As cells in the LGN mature, more low-frequency timing differences become available to the cortex, allowing DS at low frequencies to become possible for more cortical cells. PMID- 11923460 TI - Completing the corticofugal loop: a visual role for the corticogeniculate type 1 metabotropic glutamate receptor. AB - The way in which the brain deals with sensory information relies not only on feedforward processing of signals from the periphery but also on feedback inputs. This is the case of the massive projection back from layer 6 in the visual cortex to the thalamus, for which, despite being the greatest single source of synaptic contacts, the functional role still remains unclear. In the cat lateral geniculate nucleus, part of this cortical feedback is mediated by type 1 metabotropic glutamate receptors (mGluR1s), which are exclusively located on distal segments of the relay-cell dendrites. Here we show that in adult cats the cortex uses a synaptic drive mediated by these receptors (mGluR1) specifically to enhance the excitatory center of the thalamic receptive field. Moreover the effect is maximum in response to those stimuli that effectively drive cortical cells, and importantly, it does not affect the spatiotemporal structure of the thalamic receptive field. Therefore, cortex, by closing this corticofugal "loop," is able to increase the gain of its thalamic input within a focal spatial window, selecting key features of the incoming signal. PMID- 11923461 TI - Activity patterns in a model for the subthalamopallidal network of the basal ganglia. AB - Based on recent experimental data, we have developed a conductance-based computational network model of the subthalamic nucleus and the external segment of the globus pallidus in the indirect pathway of the basal ganglia. Computer simulations and analysis of this model illuminate the roles of the coupling architecture of the network, and associated synaptic conductances, in modulating the activity patterns displayed by this network. Depending on the relationships of these coupling parameters, the network can support three general classes of sustained firing patterns: clustering, propagating waves, and repetitive spiking that may show little regularity or correlation. Each activity pattern can occur continuously or in discrete episodes. We characterize the mechanisms underlying these rhythms, as well as the influence of parameters on details such as spiking frequency and wave speed. These results suggest that the subthalamopallidal circuit is capable both of correlated rhythmic activity and of irregular autonomous patterns of activity that block rhythmicity. Increased striatal input to, and weakened intrapallidal inhibition within, the indirect pathway can switch the behavior of the circuit from irregular to rhythmic. This may be sufficient to explain the emergence of correlated oscillatory activity in the subthalamopallidal circuit after destruction of dopaminergic neurons in Parkinson's disease and in animal models of parkinsonism. PMID- 11923462 TI - Role of dopamine D2-like receptors in cocaine self-administration: studies with D2 receptor mutant mice and novel D2 receptor antagonists. AB - Dopamine receptor subtypes have been classified generally as D1-like (e.g., D1, D5) or D2-like (D2, D3, D4), and converging evidence suggests that D2-like receptors may be especially important in mediating the abuse-related effects of cocaine. However, it has been difficult to differentiate the roles of the D2-like receptor subtypes in the behavioral effects of cocaine because of the relatively low selectivity of drugs for D2, D3, and D4 receptors in vivo. The goal of the present series of studies was to investigate the contributions of D2-like receptor subtypes in the reinforcing effects of cocaine using new genetic and pharmacological tools. First, we evaluated cocaine self-administration behavior, and related effects of nonselective D2-like drugs, in mutant mice that lack the D2 receptor but express D3 and D4 receptors. When high doses of cocaine on the descending limb of the cocaine dose-effect function were available, D2 mutant mice self-administered at higher rates than their heterozygous or wild-type littermates, but the ascending limb of the cocaine dose-effect function did not differ between genotypes. Elevated rates of drug intake were not attributable to nonspecific increases in response rate, because response rates maintained by presentation of a range of food concentrations were significantly lower in D2 mutant mice than in wild-type mice. In wild-type mice, pretreatment with the D2 like antagonist eticlopride increased rates of self-administration of high doses of cocaine, and the D2-like agonist quinelorane served as a positive reinforcer when substituted for cocaine. However, these effects of eticlopride and quinelorane were not observed in mice that lacked the D2 receptor. Next, we compared the effects of novel antagonists selective for different D2 receptor subtypes on cocaine self-administration behavior in outbred rats. In rats, a D2 selective antagonist increased rates of self-administration of high doses of cocaine and also combinations of cocaine and the D2-like agonist quinelorane, whereas D3/D4 antagonists were ineffective. Collectively, these findings suggest that the D2 receptor is not necessary for cocaine self-administration, but this receptor subtype is involved in mechanisms that limit rates of high-dose cocaine self-administration. Our results also suggest that D3 and D4 receptors do not play major roles in the modulation of cocaine self-administration by D2-like drugs. PMID- 11923463 TI - Evaluation of ligand-dependent changes in AR structure using peptide probes. AB - Mutations in the AR are frequently found in relapsed prostate cancers, some of which permit antiandrogens as well as nonandrogenic compounds to function as androgens. However, the molecular mechanism(s) by which these mutations enable this aberrant AR pharmacology is still unknown. To explore this issue, we used a series of LxxLL-containing peptides (L, leucine; x, any amino acid) to probe the conformation of the AF-2/coactivator binding pocket of AR and AR mutants when complexed with different ligands. We have identified in a previous study two peptides that bind to the wild-type AR in an agonist-dependent manner. Interestingly, we found these same peptides also interacted with several AR variants that are frequently found in antihormone refractory prostate cancers, in the presence of either androgens or antiandrogens. This suggests that the agonist activity of antiandrogens and other physiologically relevant ligands occurs because they, in the background of these mutations, allow AR-AF2 to adopt an active conformation. Initially, this result appeared to contradict the findings of others that suggest that coactivator binding to AR-AF2 is not required for AR activity. In probing this paradox further, however, we determined that the role of AR-AF2 appears to be to stabilize the overall structure of the receptor, allowing the amino terminus to interact with appropriate coactivators. This conclusion is supported by our finding that overexpression of the AF2-binding peptides blocks the interaction between the amino and carboxyl termini of AR but does not attenuate AR transcriptional activity. This can be explained by the fact that overexpression of the LxxLL-containing peptide or the amino terminus of AR appears to have a similar effect on the AR-ligand binding domain, as both have the ability to stabilize agonist binding by decreasing ligand off-rate. Thus, we believe that resistance in certain prostate cancers occurs as a consequence of receptor mutations that enable antagonist-and/or nonclassical ligand-bound AR to present a wild-type-like AF-2 conformation. PMID- 11923464 TI - The amino terminus of the human AR is target for corepressor action and antihormone agonism. AB - Antiandrogens inhibit the ligand-induced transactivation by the androgen receptor (AR) and have a widespread use in the treatment of prostate cancer but their mode of action is not fully understood. Here we show that the ability of the antiandrogen cyproterone acetate (CPA) to inhibit transactivation by the human AR (hAR) involves the corepressor SMRT (silencing mediator for retinoic acid and thyroid hormone receptor). We detect binding of SMRT to hAR when treating with the antiandrogen CPA, but not with the antihormones casodex or hydroxyflutamide. Interestingly, we find that SMRT binds to the N terminus of the hAR. Thereby, SMRT modulates the activity of hAR in receptor-negative CV1 cells. In addition, we have used receptor point mutants that exhibit normal transactivation potential and unchanged partial agonistic activity when treated with CPA, but lack both SMRT binding and SMRT-mediated inhibition of CPA-bound AR. This indicates that mechanisms involved in hAR-mediated transactivation are distinct from antihormone induced receptor inactivation. Furthermore, we show that treatment of transfected cells with a cAMP analog or coexpression of the catalytic subunit of PKA, known to activate hAR, inhibits the binding of SMRT to the AR. This suggests that the association of SMRT with hAR is regulated at the level of cross-talk mechanisms and that ligand-independent receptor activation is due to corepressor dissociation. Taken together, we provide novel insights in AR regulation, antihormone action, and functional nuclear receptor-corepressor interaction. PMID- 11923465 TI - The effects of estrogen-responsive element- and ligand-induced structural changes on the recruitment of cofactors and transcriptional responses by ER alpha and ER beta. AB - Estrogen signaling is mediated by ER alpha and -beta. ERs are converted from an inactive form to a transcriptionally active state through conformational changes induced by ligand and estrogen-responsive element (ERE) sequences. We show here that ER alpha and ER beta bind to an ERE independently from ER ligands. We found that although the binding affinity of ER beta for an ERE is 2-fold lower than that of ER alpha, both ERs use the same nucleotides for DNA contacts. We show that both EREs and ligands are independent modulators of ER conformation. Specifically, the ERE primarily determines the receptor-DNA affinity, whereas the structure of the ER ligand dictates the affinity of ER for particular cofactors. We found that the ligand-dependent cofactor transcriptional intermediary factor 2, through a distinct surface, also interacts with ER alpha preferentially and independently of ligand. The extent of interaction, however, is dependent upon the ER-ERE affinity. In transfected cells, ER alpha is more transcriptionally active than ER beta. The ERE sequence, however, determines the potency of gene induction when either ER subtype binds to an agonist. Antagonists prevent ERs from inducing transcription independently from ERE sequences. Thus, ERE- and ligand-induced structural changes are independent determinants for the recruitment of cofactors and transcriptional responses. The ability of ER alpha to differentially recruit a cofactor could contribute to ER subtype-specific gene responses. PMID- 11923466 TI - The presence of both the amino- and carboxyl-terminal domains in the AR is essential for the completion of a transcriptionally active form with coactivators and intranuclear compartmentalization common to the steroid hormone receptors: a three-dimensional imaging study. AB - To clarify the physiological significance of the intranuclear speckled distribution, or foci formation, of liganded steroid receptors, the subnuclear distribution of green (GFP), yellow (YFP), and cyan (CFP) fluorescent protein tagged receptors and coactivators was investigated. The foci formation of 5 alpha dihydrotestosterone (DHT)-bound AR-GFP in COS7 cells was abolished by the cotransfection of a CBP Delta (118-2393) fragment eliciting a dominant negative effect on the transactivation capacity of the AR. The N-terminal AR fragment (AR AF-1-YFP), which has a strong constitutive transactivation function, formed foci without DHT, whereas the C-terminal AR fragment (AR-AF-2-CFP), which has a quite low transactivation function, was distributed homogeneously even in the presence of DHT. The reporter gene assay showed a synergism between the transactivation functions of AR-AF-1 and AR-AF-2. This synergism was not reflected by the above two-dimensional imaging. In contrast, a three-dimensional imaging method clearly showed a difference in the intranuclear spatial distribution. The DHT-bound wild type AR-GFP alone or AR-AF-1-YFP plus DHT-bound AR-AF-2-CFP was distributed as approximately 300 discrete spots in one nucleus, whereas AR-AF-1-YFP alone was distributed as one volume in a reticular pattern. Furthermore, not only AR but also the glucocorticoid receptor-YFP, ER alpha -GFP, and YFP-tagged SRC-1, TIF2, and CBP were found to be accumulated in identical spots in the presence of ligand. All of the above results indicate that CBP is one of the factors essential for foci formation of the AR, and may propose the hypothesis that transcriptionally activated steroid receptors, regardless of the type of receptor, are transferred to common compartments (foci) and form a complex with coactivators, and this process is essential to full transactivation. PMID- 11923467 TI - Selective intranuclear redistribution of PPAR isoforms by RXR alpha. AB - The intracellular localization of transcriptionally active green fluorescent protein (GFP) chimeras linked to PPARs for human PPAR alpha (GFP-PPARh alpha) and mouse PPAR alpha, beta, and gamma 1 (GFP-PPARm alpha, GFP-PPARm beta, and GFP PPARm gamma, respectively) was examined in the mouse hepatoma cell line, Hepa-1, using fluorescence microscopy. A predominantly nuclear and diffuse distribution of each isoform was found in both the presence and absence of specific ligands for each receptor. GFP-PPARm alpha-G (containing a Glu282Gly substitution of PPARm alpha) and a phosphorylation mutant, GFP-PPARm gamma-A (containing a Ser82Ala substitution of PPARm gamma), exhibited altered transcriptional activities, but displayed similar intracellular localization patterns compared with their respective wild-type receptors. Coexpression of nuclear receptor corepressor suppressed, whereas steroid receptor coactivator-1 enhanced the transcriptional activity of each of the GFP-PPAR isoforms, but did not discernibly alter their intracellular distributions, both in the presence and absence of PPAR ligands. Interestingly, coexpression of the obligate heterodimeric partner of PPARs, RXR alpha, resulted in an intranuclear redistribution of the GFP-PPARm gamma isoform characterized by a reticulated pattern of the green fluorescent label for PPAR gamma within the nucleus, but not in nucleoli, and a heightened concentration of the fluorescent label surrounding nucleolar structures and at the nuclear membrane. Conversely, coexpression of yellow fluorescent protein-RXR alpha and native PPARm gamma resulted in a similar distribution of the yellow fluorescent tag. This localization pattern was not discernibly altered by PPAR gamma or RXR alpha-specific ligands. These results implicate RXR alpha in the nuclear reorganization of PPAR gamma and suggest that PPAR gamma colocalizes with RXR alpha at specific locations within the nucleus independent of added ligand. PMID- 11923469 TI - Lysine 183 and glutamic acid 157 of the TSH receptor: two interacting residues with a key role in determining specificity toward TSH and human CG. AB - A naturally occurring mutation in the ectodomain of the TSH receptor (TSHr), K183R, has been described recently in a familial case of gestational hyperthyroidism. Hyperthyroidism was explained by the widening of the specificity of the mutant receptor toward human CG (hCG). In the present study, we attempted to understand in molecular terms the structure-phenotype relationships of this mutant in light of the available structural model of TSHr ectodomain established on the template of the atomic structure of the porcine ribonuclease inhibitor. To this aim, we studied by site-directed mutagenesis and functional assays in transfected COS cells the effects of substituting amino acids with different physicochemical properties for lysine 183. Unexpectedly, all TSHr mutants displayed widening of their specificity toward hCG. Molecular dynamics simulations suggested that the gain of function would be secondary to the release of a nearby glutamate residue (E157) from a salt bridge with K183. This hypothesis was supported by further site-directed mutagenesis experiments showing that the presence of an acidic residue in position 157, or in its vicinity, was required to observe the increase in sensitivity to hCG (an acidic residue in position 183 can partially fulfill the role of a free acidic residue in position 157 when tested on the background of a E157A mutant). Our results suggest also that additional natural mutations (especially K183M, N, or Q) in position 183 of TSHr are expected to be found in gestational hyperthyroidism. PMID- 11923470 TI - Activation of the cAMP pathway by the TSH receptor involves switching of the ectodomain from a tethered inverse agonist to an agonist. AB - Several lines of evidence indicate that constraining intramolecular interactions between transmembrane domains are required to maintain G protein-coupled receptors in an inactive conformation in the absence of agonist. For the glycoprotein hormone receptors, which harbor a long amino-terminal ectodomain responsible for hormone binding, it has been suggested that the ectodomain could contribute to these negative constraints. To test this hypothesis, we expressed at the surface of COS-7 cells mutants of the TSH receptor in which variable portions of the amino-terminal ectodomain are replaced by a 19-residue tag from bovine rhodopsin. Whereas none of the rhodopsin-tagged truncated mutants could be activated by saturating concentrations of TSH, the constructs with the shortest amino-terminal extension displayed increased constitutive activity toward the cAMP pathway, when compared with the wild-type holoreceptor. The shortest truncated construct was strongly activated by the introduction of mutations in transmembrane segment VI (D633A), or in the third intracellular loop (A623I) of the receptor. The magnitude of the stimulation was similar to that observed when the same mutations were introduced in the intact wild-type receptor. On the contrary, the shortest truncated construct was unaffected by activating mutations affecting residues of the extracellular loop region (I486F, I568T) or the top of transmembrane segment VII (del658-661). Together, our results are compatible with a model in which activation of the cAMP pathway by the TSH receptor involves switching of the ectodomain from a tethered inverse agonist to a true agonist. PMID- 11923472 TI - Dax1 expression is dependent on steroidogenic factor 1 in the developing gonad. AB - The nuclear hormone receptor DAX1 has been implicated in mammalian gonad development and sex determination. The expression of the gene in the gonad follows a dynamic pattern in time and place in the embryo and the adult. We have undertaken the first in vivo study of the regulation of Dax1 expression. Using a transgenic mouse approach we have identified a novel 500-bp region 4 kb upstream of the mouse Dax1 start codon that is essential for LacZ reporter gene expression in the embryonic gonad. Within this region, a highly conserved steroidogenic factor 1 (SF1) consensus-binding site is necessary to direct LacZ expression to the embryonic gonad implicating SF1 in the regulation of Dax1 in the developing gonad. Consistent with this, Dax1 is expressed at much reduced levels in gonads of embryos that are deficient in SF1. In addition, our results show that SF1 consensus-binding sites close to the start of Dax1 transcription are important in regulating levels of expression in the developing gonad. These studies have identified the critical in vivo regulatory region for expression of Dax1 in the early gonad and provide novel information on how a specific enhancer element acts in different cell types at different stages of development. PMID- 11923473 TI - The nuclear receptor coactivators p300/CBP/cointegrator-associated protein (p/CIP) and transcription intermediary factor 2 (TIF2) differentially regulate PKA-stimulated transcriptional activity of steroidogenic factor 1. AB - Steroidogenic factor-1 (SF-1) is a member of the nuclear receptor superfamily that plays essential roles in the development of endocrine organs. Steroid receptor coactivator 1 and transcription intermediary factor 2 (TIF2) belong to the p160 coactivator family that mediates transcriptional activation by several nuclear receptors, including SF-1. Here, it is reported that another of the p160 coactivators, p/CIP, interacts with SF-1 through the activation function-2 domain. Both p300/CBP/cointegrator-associated protein (p/CIP) and TIF2 potentiated SF-1-mediated transcription from two reporter gene constructs in transfected nonsteroidogenic COS-1 cells and in adrenocortical Y1 cells. PKA was shown to stimulate SF-1 transcriptional activity, and coexpression of p/CIP together with the PKA catalytic subunit stimulated SF-1-mediated transactivation even further. In contrast, PKA catalytic subunit overexpression impaired the ability of TIF2 to potentiate SF-1-dependent transcription. Activation of PKA also inhibited the TIF2-mediated coactivation of other nuclear receptors such as PPAR alpha/-gamma and liver X receptor-alpha. The TIF2 mRNA levels were not affected by PKA, but instead we found that PKA activation led to a decrease in the levels of TIF2 protein. Moreover, the C-terminal activation domain 2 of TIF2 was required for the inhibitory effect of PKA, suggesting that this region is the target for the PKA-mediated down-regulation. Thus, in contrast to the regulation of p/CIP and steroid receptor coactivator 1, we suggest that activation of PKA leads to selective down-regulation of TIF2 and subsequently repression of TIF2 coactivator function. PMID- 11923474 TI - PRL activates the cyclin D1 promoter via the Jak2/Stat pathway. AB - PRL promotes cell growth and differentiation in the mammary gland, which has implications for breast cancer as well as normal development. Our data demonstrate that PRL significantly increases proliferation of mammary carcinoma cells. PRL also increases cyclin D1 levels 2-fold, which can be inhibited by actinomycin D, suggesting that transcriptional increases in cyclin D1 are important. Using a defined Chinese hamster ovary cell model system, we demonstrate that the activity of a cyclin D1 promoter-luciferase construct increases after PRL treatment. Furthermore, this increase in promoter activity is predominantly mediated by the Jak2/Stat5 signaling pathway. The cyclin D1 promoter contains two consensus sequences for PRL-induced Stat binding (GAS sites). Disruption of Stat binding to the distal GAS site destroys PRL-induced promoter activity, whereas disruption of the proximal site has no effect. We have shown by EMSA that PRL induces Stat5a and 5b to bind to the distal GAS site, and immunoprecipitation and subsequent Western analysis of nuclear extracts from PRL treated cells indicate that Stat5a and 5b can interact as a heterodimer in this system. These data suggest that cyclin D1 may be a target gene for PRL in normal lobuloalveolar development, as well as in the development and/or progression of mammary cancer. PMID- 11923475 TI - Transcriptional regulation of the human PRL-releasing peptide (PrRP) receptor gene by a dopamine 2 Receptor agonist: cloning and characterization of the human PrRP receptor gene and its promoter region. AB - PRL-releasing peptide receptor (PrRPR) mRNA was expressed in pituitary adenomas but was not detected in patients treated with bromocriptine, a specific agonist of dopamine 2 (D2) receptor. Although medical treatment with bromocriptine is effective for patients with pituitary adenomas, little is known about the molecular mechanisms of gene regulation mediated by D2 receptors. The cloned human PrRPR gene spanned approximately 2.0 kb and contained two exons and one intron. Two functional polyadenylation signals located at 510 and 714 bp downstream from the stop codon. A primer extension analysis demonstrated two major transcriptional start sites at 139 and 140 bp upstream from the translational start site and an additional minor site at -161. The promoter region contained several putative binding sites for transcriptional factors including pituitary-specific transcription factor (Pit 1), activator protein 1 (AP-1), and specificity protein (Sp1), but no typical TATA or CAAT box. This promoter showed the strong activity in the pituitary-derived GH4C1 cells, and the region between -697 and -596 bp was responsible for the stimulation both by forskolin and overexpression of cAMP response element binding protein (CREB). These stimulations were significantly suppressed by incubation with bromocriptine in a dose- and time-dependent manner, and the mutant CREB (S133A) completely abolished the inhibitory events of bromocriptine. However, EMSA studies demonstrated that CREB did not bind to this region, to which an approximately 60 kDa protein was strongly bound, and that antibodies against CREB, c-Fos, and Sp1 did not supershift this complex. Furthermore, the amount of this unknown protein was apparently reduced by treatment with bromocriptine. A series of mutation analyses demonstrated that the specific sequence, 5'-cccacatcat-3', was required for both the binding to the 60-kDa protein and the repression by bromocriptine. Therefore, the transcriptional repression of the PrRPR gene by bromocriptine required CREB but was independent of direct binding of CREB to the gene and that the sequence -663 -- -672, 5'-cccacatcat-3', bound to the 60-kDa protein appeared to be critical for this event. PMID- 11923476 TI - Variant amino acids in the extracellular loops of murine and human vasopressin V2 receptors account for differences in cell surface expression and ligand affinity. AB - Cloning and sequencing of the murine chromosomal region XB harboring the murine vasopressin V(2) receptor (mV(2)R) gene and comparison with the orthologous human Xq28 region harboring the human vasopressin V(2) receptor (hV(2)R) revealed conservation of the genomic organization and a high degree of sequence identity in the V(2)R coding regions. Despite an identity of 87% of the amino acid sequences, both receptors show marked functional differences upon stable expression in Chinese hamster ovary cells: the mV(2)R displayed a 5-fold higher affinity for [(3)H]AVP than the human ortholog; similar differences were found for the AVP-mediated activation of adenylyl cyclase. Saturation binding experiments with transiently transfected intact COS.M6 cells showed that the mV(2)R was 3- to 5-fold less abundantly expressed at the cell surface than the hV(2)R. Laser scanning microscopy of fusion proteins consisting of the V(2)Rs and green fluorescent protein (GFP) (mV(2)R/GFP, hV(2)R/GFP) demonstrated that the hV(2)R/GFP was efficiently transported to the plasma membrane, whereas the mV(2)R/GFP was localized mainly within the endoplasmic reticulum. Chimeric hV(2)Rs, in which the first and/or second extracellular loop(s) were replaced by the corresponding loop(s) of the mV(2)R, revealed that the second extracellular loop accounts for the differences in ligand binding, but the first extracellular loop accounts for the reduced cell surface expression. The exchange of lysine 100 by aspartate in the first extracellular loop of hV(2)R was sufficient to reduce cell surface expression, which was accompanied by intracellular retention as observed in laser scanning microscopy analysis. Conversely, the exchange of aspartate 100 by lysine in the mV(2)R increased the cell surface expression and resulted in predominant plasma membrane localization. Thus, a single amino acid difference in the first extracellular loop between mV(2)R and hV(2)R determines the efficiency of cell surface expression. PMID- 11923477 TI - Lysine 270 in the third intracellular domain of the oxytocin receptor is an important determinant for G alpha(q) coupling specificity. AB - To identify structural elements important to specific G alpha(q) coupling in the oxytocin receptor (OTR), intracellular domains were exchanged between OTR and G alpha(s)-coupled vasopressin V(2) receptors (V(2)Rs). Substitution of sequence from the second (2i) and third (3i) intracellular domains of V(2)R into comparable positions in OTR markedly reduced ligand affinity and resulted in a loss of G alpha(q) coupling. Substitution of the 2i domain of OTR into V(2)R decreased ligand affinity and vasopressin-stimulated adenylyl cyclase activity and only slightly increased phosphatidylinositide turnover. In contrast, substitution of the OTR3i domain into V(2)R produced a receptor chimera with high ligand affinity, decreased vasopressin-stimulated adenylyl cyclase activity, and markedly enhanced ligand-stimulated phosphatidylinositide turnover. The C terminal 36 amino acids, but not the N-terminal 13 amino acids, of the OTR3i domain contained the determinants critical for enhanced activation of PLC. Mutation of a single lysine in the C-terminal OTR3i sequence to the corresponding V(2)R residue (valine) eliminated the enhanced ability of the V(2)R chimera to stimulate PLC but did not affect maximal adenylyl cyclase stimulation. Furthermore, mutation of this residue (K270) in wild-type OTR completely abolished the ability of the receptor to stimulate phosphatidylinositide turnover, with only a small reduction in ligand affinity. These data demonstrate that OTR K270 is critically important in the stimulation by OTR of phosphatidylinositide turnover and that this determinant can also increase this activity in the V(2)R chimera. Mutation of K270 also adversely affects the ability of OTR to stimulate ERK1/2 phosphorylation. Therefore, this residue plays an important role in the specificity of OTR/G alpha(q)/PLC coupling. PMID- 11923478 TI - Angiotensinogen gene expression is dependent on signal transducer and activator of transcription 3-mediated p300/cAMP response element binding protein-binding protein coactivator recruitment and histone acetyltransferase activity. AB - Angiotensin II, a potent vasoactive peptide produced by proteolysis of the angiotensinogen (AGT) prohormone, plays a critical role in cardiovascular homeostasis. Recently we showed that IL-6 induces human (h)AGT transcription by activating the signal transducers and activators of transcription (STATs). Here we investigated the role of the coactivator p300/cAMP response element binding protein-binding protein (CBP) in STAT3-mediated hAGT gene expression. Overexpression of adenovirus 12S E1A, which binds and inactivates p300/CBP, strongly inhibited basal and stimulated hAGT transcription, whereas a mutant E1A defective in binding p300/CBP did not. Conversely, ectopic expression of p300 and CBP potentiated inducible hAGT promoter activity. Coimmunoprecipitation assays revealed STAT3-p300 interaction upon IL-6 stimulation. The STAT3-p300 association requires the STAT3 C-terminal transactivation domain, as STAT3 deleted of transactivation functions as a dominant-negative inhibitor and does not associate with p300/CBP. The observation that IL-6 stimulation increases histone H4 acetylation of the endogenous hAGT promoter, and expression of p300 deficient in histone acetyltransferase activity down-regulates hAGT promoter activity both suggest that p300 histone acetyltransferase activity is required for hAGT expression. Finally, treatment of HepG2 cells with a histone deacetylase inhibitor increased the hAGT mRNA abundance by 2- to 3-fold. Taken together, our results indicate that IL-6-inducible expression of the hAGT promoter is mediated by physical association of the COOH terminus of STAT3 with p300/CBP, the recruitment of which targets histone acetylation and results in chromatin remodeling. PMID- 11923479 TI - The synergistic activity of thyroid transcription factor 1 and Pax 8 relies on the promoter/enhancer interplay. AB - The transcription factors, thyroid transcription factor 1 (TTF-1) and Pax 8, play a pivotal role in the transcriptional regulation of the thyroid differentiation marker genes and in the differentiation of the thyroid follicular cells. They have a very restricted tissue distribution, and the thyrocyte is the only cell type with the simultaneous expression of these factors. Here we show that TTF-1 and Pax 8 cooperatively activate their target genes and that their synergistic activity requires the cross-talk between enhancer and gene promoter. We have characterized the cis and trans requirements of the TTF1/Pax 8 synergistic activity on the thyroperoxidase gene. We show that their synergy is also important for thyroglobulin gene transcription. PMID- 11923480 TI - Knockout of PKC alpha enhances insulin signaling through PI3K. AB - Insulin stimulates glucose transport and certain other metabolic processes by activating atypical PKC isoforms (lambda, zeta, iota) and protein kinase B (PKB) through increases in D3-polyphosphoinositides derived from the action of PI3K. The role of diacylglycerol-sensitive PKC isoforms is less clear as they have been suggested to be both activated by insulin and yet inhibit insulin signaling to PI3K. Presently, we found that insulin signaling to insulin receptor substrate 1 dependent PI3K, PKB, and PKC lambda, and downstream processes, glucose transport and activation of ERK, were enhanced in skeletal muscles and adipocytes of mice in which the ubiquitous conventional diacylglycerol-sensitive PKC isoform, PKC alpha, was knocked out by homologous recombination. On the other hand, insulin provoked wortmannin-insensitive increases in immunoprecipitable PKC alpha activity in adipocytes and skeletal muscles of wild-type mice and rats. We conclude that 1) PKC alpha is not required for insulin-stimulated glucose transport, and 2) PKC alpha is activated by insulin at least partly independently of PI3K, and largely serves as a physiological feedback inhibitor of insulin signaling to the insulin receptor substrate 1/PI3K/PKB/PKC lambda/zeta/iota complex and dependent metabolic processes. PMID- 11923481 TI - Identification of the critical sequence elements in the cytoplasmic domain of leptin receptor isoforms required for Janus kinase/signal transducer and activator of transcription activation by receptor heterodimers. AB - Two predominant splice variants of the leptin receptor (LEPR) are coexpressed in leptin-responsive tissues: the long form, LEPRb, characterized as the signal transducing receptor, and the signaling-defective short form, LEPRa. It is unknown whether heterodimers of these isoforms are capable of signal transduction via the Janus kinase (JAK)/signal transducer and activator of transcription (STAT) pathway. To address this question, chimeric receptors were constructed consisting of the transmembrane and intracellular parts of LEPRb and LEPRa fused with the extracellular domains of either the alpha- or beta-subunit of the IL-5 receptor. This strategy allows the directed heterodimerization of different LEPR cytoplasmic tails and excludes homodimerization. In COS-7 and HEPG2 cells, chimeric receptor heterodimers of LEPRa and LEPRb failed to activate the JAK/STAT pathway, whereas receptor dimers of LEPRb gave rise to the expected ligand dependent activation of JAK2, phosphorylation of STAT3, and STAT3-dependent promoter activity. Markedly lower amounts of JAK2 were found to be associated with immunoprecipitated LEPRa chimeras than with LEPRb chimeras. Analysis of a series of deletion constructs indicated that a segment of 15 amino acids in addition to the 29 amino acids common to LEPRa and LEPRb was required for partial restoration of JAK/STAT activation. Site-directed mutagenesis of the critical sequence indicated that two hydrophobic residues (Leu896, Phe897) not present in LEPRa were indispensable for receptor signaling. These findings show that LEPRa/LEPRb heterodimers cannot activate STAT3 and identify sequence elements within the LEPR that are critical for the activation of JAK2 and STAT3. PMID- 11923483 TI - Disturbances in glucose-tolerance, insulin-release, and stress-induced hyperglycemia upon disruption of the Ca(v)2.3 (alpha 1E) subunit of voltage-gated Ca(2+) channels. AB - Multiple types of voltage-activated Ca(2+) channels (T, L, N, P, Q, R type) coordinate Ca(2+)-dependent processes in neurons and neuroendocrine cells. Expressional and functional data have suggested a role for Ca(v)2.3 Ca(2+) channels in endocrine processes. To verify its role in vivo, Ca(v)2.3(-/-) mutant mice were generated, thus deficient in alpha 1E/R-type Ca(2+) channel. Intraperitoneal injection of D-glucose showed that glucose tolerance was markedly reduced, and insulin release into plasma was impaired in Ca(v)2.3-deficient mice. In isolated islets of Langerhans from these animals, no glucose-induced insulin release was detected. Further, in stressed Ca(v)2.3-deficient mice, the rate of glucose release into the blood was only 29% of that observed for wild-type animals. Thus, the deletion of Ca(v)2.3 causes deficits not only in insulin release but also in stress-induced hyperglycemia. The complex phenotype of Ca(v)2.3-deficient mice has dual components related to endocrine and neurological defects. The present findings provide direct evidence of a functional role for the Ca(v)2.3 subunit in hormone secretion and glucose homeostasis. PMID- 11923484 TI - Endocrine-related resources from the National Institutes of Health. PMID- 11923482 TI - Insulin and TNF alpha induce expression of the forkhead transcription factor gene Foxc2 in 3T3-L1 adipocytes via PI3K and ERK 1/2-dependent pathways. AB - We have recently identified the winged helix/forkhead gene Foxc2 as a key regulator of adipocyte metabolism that counteracts obesity and diet-induced insulin resistance. This study was performed to elucidate the hormonal regulation of Foxc2 in adipocytes. We find that TNF alpha and insulin induce Foxc2 mRNA in differentiated 3T3-L1 cells with the kinetics of an immediate early response (1-2 h with 100 ng/ml insulin or 5 ng/ml TNF alpha). This induction is, in both cases, attenuated by the PI3K inhibitor wortmannin as well as the MAPK kinase inhibitor PD98059. Furthermore, we show that stimulation of 3T3-L1 adipocytes with phorbol 12-myristate-13-acetate or 8-(4-chlorophenyl)thio-cAMP induces the expression of Foxc2. Interestingly, we find that the basal level of Foxc2 mRNA is down regulated whereas hormonal responsiveness increases during differentiation of 3T3 L1 from preadipocytes to adipocytes. At the protein level, immunoblots with Foxc2 antibody demonstrated an induction of Foxc2 by insulin and TNF alpha in nuclear extracts of 3T3-L1 adipocytes. EMSA of nuclear proteins from phorbol-12-myristate 13-acetate- and TNF alpha-treated 3T3-L1 adipocytes using a forkhead consensus oligonucleotide revealed specific binding of a Foxc2/DNA complex. In conclusion, our data suggest that insulin and TNF alpha regulate the expression of Foxc2 via a PI3K- and ERK 1/2-dependent pathway in 3T3-L1 adipocytes. Also, signaling pathways downstream of PKA and PKC induce the expression of Foxc2 mRNA. PMID- 11923485 TI - Smallpox and bioterrorism. PMID- 11923486 TI - Smallpox, October 1945. PMID- 11923487 TI - Smallpox vaccination policy--the need for dialogue. PMID- 11923488 TI - Images in clinical medicine. Eczema vaccinatum--a timely reminder. PMID- 11923489 TI - Dose-related effects of smallpox vaccine. AB - BACKGROUND: We conducted a double-blind, randomized trial of three dilutions of vaccinia virus vaccine in previously unimmunized adults in order to assess the clinical success rates, humoral responses, and virus-specific activity of cytotoxic T cells and interferon-gamma-producing T cells. METHODS: Sixty healthy adults were inoculated intradermally by bifurcated needle with undiluted vaccine (dose, 10(7.8) plaque-forming units [pfu] per milliliter), a 1:10 dilution (dose, 10(6.5) pfu per milliliter), or a 1:100 dilution (dose, 10(5.0) pfu per milliliter); there were 20 subjects in each group. The subjects were monitored with respect to vesicle formation (an indicator of successful vaccination), the viral titer at the time of peak lesion formation, antiviral antibodies, and cellular immune responses. RESULTS: A vaccinia vesicle developed in 19 of the 20 subjects who received undiluted vaccine (95 percent), 14 of the 20 who received the 1:10 dilution (70 percent), and 3 of the 20 who received the 1:100 dilution (15 percent). One month after vaccination, 34 of 36 subjects with vesicles had antibody responses, as compared with only 1 of 24 subjects without clinical evidence of vaccinia virus replication. Vigorous cytotoxic T-cell and interferon gamma responses occurred in 94 percent of subjects with vesicles, and a cytotoxic T-cell response occurred in only one subject without a vesicle. CONCLUSIONS: The vaccinia virus vaccine (which was produced in 1982 or earlier) still has substantial potency when administered by a bifurcated needle to previously unvaccinated adults. Diluting the vaccine reduces the rate of successful vaccination. The development of vesicular skin lesions after vaccination correlates with the induction of the antibody and T-cell responses that are considered essential for clearing vaccinia virus infections. PMID- 11923490 TI - Clinical responses to undiluted and diluted smallpox vaccine. AB - BACKGROUND: To evaluate the potential to increase the supply of smallpox vaccine (vaccinia virus), we compared the response to vaccination with 10(8.1), 10(7.2), and 10(7.0) plaque-forming units (pfu) of vaccinia virus per milliliter. METHODS: In this randomized, single-blind, prospective study, 680 adults who had not been previously immunized were inoculated intradermally with undiluted vaccine (mean titer, 10(8.1) pfu per milliliter), a 1:5 dilution, or a 1:10 dilution of vaccinia virus with use of a bifurcated needle, and the site was covered with a semipermeable dressing. Subjects were monitored for vesicle formation (an indicator of the success of vaccination) and adverse events for 56 days after immunization. RESULTS: Success rates did not differ significantly among the groups and ranged from 97.1 to 99.1 percent after the first vaccination. Both the undiluted and diluted vaccines were reactogenic. In addition to the formation of pustules, common adverse events included the formation of satellite lesions, regional lymphadenopathy, fever, headache, nausea, muscle aches, fatigue, and chills consistent with the presence of an acute viral illness. Generalized and localized rashes, including two cases of erythema multiforme, were also observed. CONCLUSIONS: When given by a bifurcated needle, vaccinia virus vaccine can be diluted to a titer as low as 10(7.0) pfu per milliliter (approximately 10,000 pfu per dose) and induce local viral replication and vesicle formation in more than 97 percent of persons. PMID- 11923491 TI - Diagnosis and management of smallpox. PMID- 11923492 TI - The case for voluntary smallpox vaccination. PMID- 11923493 TI - Experimental quantum cloning of single photons. AB - Although perfect copying of unknown quantum systems is forbidden by the laws of quantum mechanics, approximate cloning is possible. A natural way of realizing quantum cloning of photons is by stimulated emission. In this context, the fundamental quantum limit to the quality of the clones is imposed by the unavoidable presence of spontaneous emission. In our experiment, a single input photon stimulates the emission of additional photons from a source on the basis of parametric down-conversion. This leads to the production of quantum clones with near-optimal fidelity. We also demonstrate universality of the copying procedure by showing that the same fidelity is achieved for arbitrary input states. PMID- 11923494 TI - Genetic dissection of transcriptional regulation in budding yeast. AB - To begin to understand the genetic architecture of natural variation in gene expression, we carried out genetic linkage analysis of genomewide expression patterns in a cross between a laboratory strain and a wild strain of Saccharomyces cerevisiae. Over 1500 genes were differentially expressed between the parent strains. Expression levels of 570 genes were linked to one or more different loci, with most expression levels showing complex inheritance patterns. The loci detected by linkage fell largely into two categories: cis-acting modulators of single genes and trans-acting modulators of many genes. We found eight such trans-acting loci, each affecting the expression of a group of 7 to 94 genes of related function. PMID- 11923496 TI - Biological weapons and international law. PMID- 11923495 TI - Alteration of lymphocyte trafficking by sphingosine-1-phosphate receptor agonists. AB - Blood lymphocyte numbers, essential for the development of efficient immune responses, are maintained by recirculation through secondary lymphoid organs. We show that lymphocyte trafficking is altered by the lysophospholipid sphingosine-1 phosphate (S1P) and by a phosphoryl metabolite of the immunosuppressive agent FTY720. Both species were high-affinity agonists of at least four of the five S1P receptors. These agonists produce lymphopenia in blood and thoracic duct lymph by sequestration of lymphocytes in lymph nodes, but not spleen. S1P receptor agonists induced emptying of lymphoid sinuses by retention of lymphocytes on the abluminal side of sinus-lining endothelium and inhibition of egress into lymph. Inhibition of lymphocyte recirculation by activation of S1P receptors may result in therapeutically useful immunosuppression. PMID- 11923498 TI - High-energy physics. Cost overruns will hit research at CERN. PMID- 11923497 TI - Advising government. DARPA and Jason divorce in spat over membership. PMID- 11923499 TI - Cosmology. Cosmic ripples confirm universe speeding up. PMID- 11923500 TI - Germany. Academic reform law made more flexible. PMID- 11923501 TI - Seismology. Deep quakes slow but very steady. PMID- 11923502 TI - Environmental fellowships. Will Congress catch EPA's falling STAR? PMID- 11923503 TI - Neuroscience. Neurons turn a blind eye to eye movements. PMID- 11923504 TI - Telomeres. Chromosome end game draws a crowd. PMID- 11923505 TI - Tackling cancer at the telomeres. PMID- 11923506 TI - Water quality. Microbiologists on the trail of polluting bacteria. PMID- 11923507 TI - Water quality. Is E. coli distinct enough to join the hunt? PMID- 11923508 TI - France. Biologist wins battle over bureaucratic fungus. PMID- 11923509 TI - Argentina. Economic crash brings ill winds for science. PMID- 11923510 TI - Entomology. Getting the behavior of social insects to compute. PMID- 11923512 TI - Paleoclimate. Ice sheet collapse and sea level change. PMID- 11923511 TI - Global genetic resources. Science and the Convention on Biological Diversity. PMID- 11923513 TI - Psychology. The effects of media violence on society. PMID- 11923514 TI - Earth sciences. The study of superfloods. PMID- 11923515 TI - Biomedicine. Defining the "S" in SERMs. PMID- 11923516 TI - Structural biology and biochemistry. Retrospective: Max Perutz (1914-2002). PMID- 11923517 TI - Neuroscience. Moving through the landscape. PMID- 11923518 TI - Materials science. Deformation of nanostructures. PMID- 11923519 TI - Matrix metalloproteinase inhibitors and cancer: trials and tribulations. AB - For at least 30 years, matrix metalloproteinases (MMPs) have been heralded as promising targets for cancer therapy on the basis of their massive up-regulation in malignant tissues and their unique ability to degrade all components of the extracellular matrix. Preclinical studies testing the efficacy of MMP suppression in tumor models were so compelling that synthetic metalloproteinase inhibitors (MPIs) were rapidly developed and routed into human clinical trials. The results of these trials have been disappointing. Here we review the studies that brought MPIs into clinical testing and discuss the design and outcome of the trials in light of new information about the cellular source, substrates, and mode of action of MMPs at different stages of tumor progression. The important lessons learned from the MPI experience may be of great value for future studies of MPIs and for cancer drug development in general. PMID- 11923521 TI - Chemists look to follow biology lead. PMID- 11923522 TI - Can chemists assemble a future for molecular electronics? PMID- 11923523 TI - Building better photonic crystals. PMID- 11923524 TI - Toward self-organization and complex matter. AB - Beyond molecular chemistry based on the covalent bond, supramolecular chemistry aims at developing highly complex chemical systems from components interacting through noncovalent intermolecular forces. Over the past quarter century, supramolecular chemistry has grown into a major field and has fueled numerous developments at the interfaces with biology and physics. Some of the conceptual advances and future challenges are profiled here. PMID- 11923525 TI - Synthesis beyond the molecule. AB - Weak, noncovalent interactions between molecules control many biological functions. In chemistry, noncovalent interactions are now exploited for the synthesis in solution of large supramolecular aggregates. The aim of these syntheses is not only the creation of a particular structure, but also the introduction of specific chemical functions in these supramolecules. PMID- 11923526 TI - Functional materials based on self-assembly of polymeric supramolecules. AB - Self-assembly of polymeric supramolecules is a powerful tool for producing functional materials that combine several properties and may respond to external conditions. We illustrate the concept using a comb-shaped architecture. Examples include the hexagonal self-organization of conjugated conducting polymers and the polarized luminance in solid-state films of rodlike polymers obtained by removing the hydrogen-bonded side chains from the aligned thermotropic smectic phase. Hierarchically structured materials obtained by applying different self organization and recognition principles and directed assembly form a basis for tunable nanoporous materials, smart membranes, preparation of nano-objects, and anisotropic properties, such as proton conductivity. PMID- 11923527 TI - Crystal engineering: from structure to function. AB - Modern crystal engineering has emerged as a rich discipline whose success requires an iterative process of synthesis, crystallography, crystal structure analysis, and computational methods. By focusing on the molecular recognition events during nucleation and growth, chemists have uncovered new ways of controlling the internal structure and symmetry of crystals and of producing materials with useful chemical and physical properties. PMID- 11923528 TI - Self-assembly of phase-segregated liquid crystal structures. AB - Additional functionality can be incorporated into liquid crystalline materials by using phase segregation and self-assembly. Intermolecular interactions such as hydrogen bonding and ionic interactions play key roles in the formation of these complex structures. One-, two-, and three-dimensional phase-segregated structures on various scales of length are formed by self-assembly of a variety of partially incompatible molecules. Such structures can enhance anisotropic properties such as ionic conductivity. PMID- 11923529 TI - Self-assembly at all scales. AB - Self-assembly is the autonomous organization of components into patterns or structures without human intervention. Self-assembling processes are common throughout nature and technology. They involve components from the molecular (crystals) to the planetary (weather systems) scale and many different kinds of interactions. The concept of self-assembly is used increasingly in many disciplines, with a different flavor and emphasis in each. PMID- 11923530 TI - Periodic slow earthquakes from the Cascadia subduction zone. PMID- 11923531 TI - Hybrid nanorod-polymer solar cells. AB - We demonstrate that semiconductor nanorods can be used to fabricate readily processed and efficient hybrid solar cells together with polymers. By controlling nanorod length, we can change the distance on which electrons are transported directly through the thin film device. Tuning the band gap by altering the nanorod radius enabled us to optimize the overlap between the absorption spectrum of the cell and the solar emission spectrum. A photovoltaic device consisting of 7-nanometer by 60-nanometer CdSe nanorods and the conjugated polymer poly 3(hexylthiophene) was assembled from solution with an external quantum efficiency of over 54% and a monochromatic power conversion efficiency of 6.9% under 0.1 milliwatt per square centimeter illumination at 515 nanometers. Under Air Mass (A.M.) 1.5 Global solar conditions, we obtained a power conversion efficiency of 1.7%. PMID- 11923532 TI - Theory of quantum annealing of an Ising spin glass. AB - Probing the lowest energy configuration of a complex system by quantum annealing was recently found to be more effective than its classical, thermal counterpart. By comparing classical and quantum Monte Carlo annealing protocols on the two dimensional random Ising model (a prototype spin glass), we confirm the superiority of quantum annealing relative to classical annealing. We also propose a theory of quantum annealing based on a cascade of Landau-Zener tunneling events. For both classical and quantum annealing, the residual energy after annealing is inversely proportional to a power of the logarithm of the annealing time, but the quantum case has a larger power that makes it faster. PMID- 11923533 TI - A phase separation model for the nanopatterning of diatom biosilica. AB - Diatoms are encased in an intricately patterned wall that consists of amorphous silica. Species-specific fabrication of this ornate biomineral enables taxonomists to identify thousands of diatom species. The molecular mechanisms that control this nanofabrication and generate the diversity of patterns is not well understood. A simple model is described, in which repeated phase separation events during wall biogenesis are assumed to produce self-similar silica patterns in smaller and smaller scales. On the basis of this single assumption, the apparently complex patterns found in the valves of the diatom genus Coscinodiscus can be predicted. Microscopic analysis of valves in statu nascendi from three different Coscinodiscus species supports the conclusions derived from the model. PMID- 11923534 TI - Atomic-level observation of disclination dipoles in mechanically milled, nanocrystalline Fe. AB - Plastic deformation of materials occurs by the motion of defects known as dislocations and disclinations. High-resolution transmission electron microscopy was used to directly reveal the individual dislocations that constitute partial disclination dipoles in nanocrystalline, body-centered cubic iron that had undergone severe plastic deformation by mechanical milling. The mechanisms by which the formation and migration of such partial disclination dipoles during deformation allow crystalline solids to fragment and rotate at the nanometer level are described. Such rearrangements are important basic phenomena that occur during material deformation, and hence, they may be critical in the formation of nanocrystalline metals by mechanical milling and other deformation processes. PMID- 11923535 TI - Orbital influence on Earth's magnetic field: 100,000-year periodicity in inclination. AB - A continuous record of the inclination and intensity of Earth's magnetic field, during the past 2.25 million years, was obtained from a marine sediment core of 42 meters in length. This record reveals the presence of 100,000-year periodicity in inclination and intensity, which suggests that the magnetic field is modulated by orbital eccentricity. The correlation between inclination and intensity shifted from antiphase to in-phase, corresponding to a magnetic polarity change from reversed to normal. To explain the observation, we propose a model in which the strength of the geocentric axial dipole field varies with 100,000-year periodicity, whereas persistent nondipole components do not. PMID- 11923537 TI - Senescence induced by altered telomere state, not telomere loss. AB - Primary human cells in culture invariably stop dividing and enter a state of growth arrest called replicative senescence. This transition is induced by programmed telomere shortening, but the underlying mechanisms are unclear. Here, we report that overexpression of TRF2, a telomeric DNA binding protein, increased the rate of telomere shortening in primary cells without accelerating senescence. TRF2 reduced the senescence setpoint, defined as telomere length at senescence, from 7 to 4 kilobases. TRF2 protected critically short telomeres from fusion and repressed chromosome-end fusions in presenescent cultures, which explains the ability of TRF2 to delay senescence. Thus, replicative senescence is induced by a change in the protected status of shortened telomeres rather than by a complete loss of telomeric DNA. PMID- 11923536 TI - Role of a ubiquitin-like modification in polarized morphogenesis. AB - Type I ubiquitin-like proteins constitute a family of protein modifiers. Here we report the identification of a posttranslational protein modifier from Saccharomyces cerevisiae, Hub1. Overexpression of Hub1 resulted in enhanced conjugate formation when its carboxyl-terminal residue was deleted, suggesting that mature Hub1 may be produced by proteolytic processing. In vivo targets of Hub1 conjugation included cell polarity factors Sph1 and Hbt1. In the hub1Delta mutant, the subcellular localization of both Hbt1 and Sph1 was disrupted, and cell polarization during the formation of mating projections was defective. Consistent with these polarization defects, the hub1Delta mutant was deficient in mating. PMID- 11923538 TI - Visualization of a Ran-GTP gradient in interphase and mitotic Xenopus egg extracts. AB - The small guanosine triphosphatase Ran is loaded with guanosine triphosphate (GTP) by the chromatin-bound guanine nucleotide exchange factor RCC1 and releases import cargoes in the nucleus during interphase. In mitosis, Ran-GTP promotes spindle assembly around chromosomes by locally discharging cargoes that regulate microtubule dynamics and organization. We used fluorescence resonance energy transfer-based biosensors to visualize gradients of Ran-GTP and liberated cargoes around chromosomes in mitotic Xenopus egg extracts. Both gradients were required to assemble and maintain spindle structure. During interphase, Ran-GTP was highly enriched in the nucleoplasm, and a steep concentration difference between nuclear and cytoplasmic Ran-GTP was established, providing evidence for a Ran-GTP gradient surrounding chromosomes throughout the cell cycle. PMID- 11923539 TI - Neural mechanisms of saccadic suppression. AB - In normal vision our gaze leaps from detail to detail, resulting in rapid image motion across the retina. Yet we are unaware of such motion, a phenomenon known as saccadic suppression. We recorded neural activity in the middle temporal and middle superior temporal cortical areas during saccades and identical image motion under passive viewing conditions. Some neurons were selectively silenced during saccadic image motion, but responded well to identical external image motion. In addition, a subpopulation of neurons reversed their preferred direction of motion during saccades. Consequently, oppositely directed motion signals annul one another, and motion percepts are suppressed. PMID- 11923540 TI - Cortical neurons encoding path and place: where you go is where you are. AB - We recorded neuronal activity in monkey medial superior temporal (MST) cortex during movement on a motorized sled. Most neurons showed a preferred heading direction, but some responded only when that heading was part of a particular path. Others responded only when the animal was at a certain place in the room, regardless of its path to that place. Video simulations of the self-movement scene evoked path, but not place, responses. Stationary positioning in the room revealed location preferences that matched place preferences recorded during movement. We conclude that MST encodes heading, path, and place information to support visuospatial orientation. PMID- 11923542 TI - Television viewing and aggressive behavior during adolescence and adulthood. AB - Television viewing and aggressive behavior were assessed over a 17-year interval in a community sample of 707 individuals. There was a significant association between the amount of time spent watching television during adolescence and early adulthood and the likelihood of subsequent aggressive acts against others. This association remained significant after previous aggressive behavior, childhood neglect, family income, neighborhood violence, parental education, and psychiatric disorders were controlled statistically. PMID- 11923543 TI - Are oral appliances a substitute for nasal positive airway pressure? PMID- 11923541 TI - Molecular determinants for the tissue specificity of SERMs. AB - Selective estrogen receptor modulators (SERMs) mimic estrogen action in certain tissues while opposing it in others. The therapeutic effectiveness of SERMs such as tamoxifen and raloxifene in breast cancer depends on their antiestrogenic activity. In the uterus, however, tamoxifen is estrogenic. Here, we show that both tamoxifen and raloxifene induce the recruitment of corepressors to target gene promoters in mammary cells. In endometrial cells, tamoxifen, but not raloxifene, acts like estrogen by stimulating the recruitment of coactivators to a subset of genes. The estrogen-like activity of tamoxifen in the uterus requires a high level of steroid receptor coactivator 1 (SRC-1) expression. Thus cell type and promoter-specific differences in coregulator recruitment determine the cellular response to SERMs. PMID- 11923544 TI - Vocal cord dysfunction and laryngeal hyperresponsiveness: a function of altered autonomic balance? PMID- 11923545 TI - British Thoracic Society Winter Meeting 2001. PMID- 11923547 TI - Can mandibular advancement devices be a satisfactory substitute for short term use in patients on nasal continuous positive airway pressure? AB - BACKGROUND: Mandibular advancement devices (MADs) can successfully control both snoring and obstructive sleep apnoea (OSA). Many patients on nasal continuous positive airway pressure (NCPAP) for OSA would like a more portable alternative, even if only temporarily. This study assesses what proportion of patients with OSA already on NCPAP can successfully use a MAD for short periods (up to 1 month) as a temporary alternative to NCPAP. METHODS: Fifty patients with OSA, already on NCPAP for at least 3 months, were recruited by invitation. They were provided with a simple fixed MAD estimated to provide 75% of maximum mandibular protrusion. Sleep studies using a portable home recorder were performed on and after three nights without NCPAP to provide control data. Following acclimatisation to the MAD, sleep studies were also planned after 3, 7, and 28 days while using the MAD. If their overnight >4% SaO(2) dips per hour deteriorated to >20 or the Epworth sleepiness score (ESS) rose to >9 (or increased by >4 over baseline) on nights 3 or 7, they were then deemed to have failed the trial and were withdrawn. RESULTS: Of the 50 patients entered, one had inadequate teeth for a MAD and 31 gave up trying to use the device during the acclimatisation period because of side effects. Of the 18 prepared to use the device, two patients failed at night 3, five at night 7, and two at night 28. Thus, nine patients remained controlled by our criteria at night 28. On average, sleep study indices while using the MAD were poor compared with the night on NCPAP. CONCLUSIONS: Simple MADs are poorly tolerated by patients with OSA already on NCPAP. OSA was adequately controlled by our criteria in 32% of those recruited for the equivalent of a weekend, in 22% for 1 week, and in 18% for up to 1 month. Better tolerated devices would be likely to improve on these figures. PMID- 11923546 TI - Managing passengers with respiratory disease planning air travel: British Thoracic Society recommendations. PMID- 11923549 TI - Case-control study of severe life threatening asthma (SLTA) in adults: psychological factors. AB - BACKGROUND: Severe life threatening asthma (SLTA) is important in its own right and as a proxy for asthma death. In order to target hospital based intervention strategies to those most likely to benefit, risk factors for SLTA among those admitted to hospital need to be identified. Adverse psychological factors are purported risk factors for asthma death and SLTA /near fatal asthma. A study was undertaken to determine whether, in comparison with patients admitted to hospital with acute asthma, those with SLTA have specific adverse psychological factors. METHODS: A case-control study was undertaken. Cases (n=77) were admitted to the intensive care unit with SLTA (mean (SD) pH 7.17 (0.15), PaCO(2) 10.7 (5.0) kPa). Controls (n=239) were admitted to general wards with acute asthma and were matched only by date of index attack. An interviewer administered questionnaire was undertaken 24-48 hours after admission. A random sample of community based asthmatics was recruited to provide normative data on asthmatics for comparison with cases and hospital controls. RESULTS: The risk of SLTA increased with age (OR 1.04/year, 95% CI 1.01 to 1.07) and was less for women (OR 0.36, 95% CI 0.20 to 0.68). These variables were controlled for in all further analyses. There was a high prevalence of psychological disorder in both cases and matched controls, but there was no difference in prevalence of caseness for anxiety or depression, total (or individual) life events in last 12 months, availability of general or disease specific social support, nor in any of the domains of the Attitudes and Beliefs about Asthma Questionnaire (emotional (mal) adjustment, doctor-patient relationship, stigma, self-efficacy). Cases (SLTA) were less likely to have had previous emotional counselling (25% v. 35%, p<0.05). However, when comparison was made with a community based group of asthmatic patients, those admitted to hospital with acute asthma (SLTA and hospital controls) had a higher prevalence of anxiety and depression, higher total life events, and higher prevalence of certain specific life events. CONCLUSIONS: There was considerable psychological morbidity generally (and anxiety specifically) in those admitted with acute asthma. Specific adverse psychological factors were not risk factors for SLTA, when comparison was made with those admitted to hospital with acute asthma, but adverse psychological factors were a risk factor for hospitalisation for acute asthma (including SLTA). Psychological risk factors for adverse events in asthma are dependent both on the type of event under study and the comparison group used. PMID- 11923548 TI - Airway inflammation, basement membrane thickening and bronchial hyperresponsiveness in asthma. AB - BACKGROUND: There are few data in asthma relating airway physiology, inflammation and remodelling and the relative effects of inhaled corticosteroid (ICS) treatment on these parameters. A study of the relationships between spirometric indices, airway inflammation, airway remodelling, and bronchial hyperreactivity (BHR) before and after treatment with high dose inhaled fluticasone propionate (FP 750 microg bd) was performed in a group of patients with relatively mild but symptomatic asthma. METHODS: A double blind, randomised, placebo controlled, parallel group study of inhaled FP was performed in 35 asthmatic patients. Bronchoalveolar lavage (BAL) and airway biopsy studies were carried out at baseline and after 3 and 12 months of treatment. Twenty two normal healthy non asthmatic subjects acted as controls. RESULTS: BAL fluid eosinophils, mast cells, and epithelial cells were significantly higher in asthmatic patients than in controls at baseline (p<0.01). Subepithelial reticular basement membrane (rbm) thickness was variable, but overall was increased in asthmatic patients compared with controls (p<0.01). Multiple regression analysis explained 40% of the variability in BHR, 21% related to rbm thickness, 11% to BAL epithelial cells, and 8% to BAL eosinophils. The longitudinal data corroborated the cross sectional model. Forced expiratory volume in 1 second improved after 3 months of treatment with FP with no further improvement at 12 months. PD(20) improved throughout the study. BAL inflammatory cells decreased following 3 months of treatment with no further improvement at 12 months (p<0.05 v placebo). Rbm thickness decreased in the FP group, but only after 12 months of treatment (mean change -1.9, 95% CI -3 to -0.7 microm; p<0.01 v. baseline, p<0.05 v. placebo). A third of the improvement in BHR with FP was associated with early changes in inflammation, but the more progressive and larger improvement was associated with the later improvement in airway remodelling. CONCLUSION: Physiology, airway inflammation and remodelling in asthma are interrelated and improve with ICS. Changes are not temporally concordant, with prolonged treatment necessary for maximal benefit in remodelling and PD(20). Determining the appropriate dose of inhaled steroids only by reference to symptoms and lung function, as specified in current international guidelines, and even against indices of inflammation may be over simplistic. The results of this study support the need for early and long term intervention with ICS, even in patients with relatively mild asthma. PMID- 11923550 TI - Role of cysteinyl leukotrienes in adenosine 5'-monophosphate induced bronchoconstriction in asthma. AB - BACKGROUND: Adenosine induced bronchoconstriction in patients with asthma is thought to be mediated by the synthesis and release of autacoids from airway mast cells. In vitro, adenosine induced constriction of asthmatic bronchi is blocked by a combination of specific histamine and cysteinyl leukotriene receptor antagonists, but the relative contribution of these mediators in vivo is unclear. We hypothesised that adenosine induced bronchoconstriction in asthmatic patients may be blocked by pretreatment with the orally active selective cysteinyl leukotriene-1 (CysLT(1)) receptor antagonist, montelukast. METHODS: In a randomised, double blind, crossover study, oral montelukast (10 mg) or placebo was administered once daily on two consecutive days to 18 patients with mild to moderate persistent atopic asthma. Incremental doses of adenosine 5' monophosphate (AMP) from 0.39 to 400 mg/ml were inhaled by dosimeter and the dose producing a 20% fall in FEV(1) (PC(20)AMP) after AMP inhalation was recorded. Leukotriene E(4) (LTE(4)) urinary concentrations were measured by enzyme immunoassay 4 hours after AMP challenge. RESULTS: Montelukast pretreatment provided highly significant protection against adenosine induced bronchoconstriction, with geometric mean PC(20)AMP values of 52.6 mg/ml (95% CI 35.2 to 78.7) after placebo and 123.9 mg/ml (95% CI 83.0 to 185.0) after montelukast (p=0.006). The geometric mean of the montelukast/placebo PC(20)AMP ratio was 2.4 (95% CI 1.3 to 4.2). Montelukast had no significant effect on 4 hour urinary excretion of LTE(4) compared with placebo. CONCLUSIONS: Selective CysLT(1) receptor antagonism with montelukast provides highly significant protection against AMP induced bronchoconstriction in patients with atopic asthma, implying that cysteinyl leukotrienes are generated from airway mast cells through preferential activation of their A(2B) receptors. PMID- 11923551 TI - A defective type 1 response to rhinovirus in atopic asthma. AB - BACKGROUND: Rhinoviruses (RVs) are the most frequent precipitants of the common cold and asthma exacerbations, but little is known about the immune response to these viruses and its potential implications in the pathogenesis of asthma. METHODS: Peripheral blood mononuclear cells (PBMC) from patients with atopic asthma and normal subjects were exposed to live or inactivated RV preparations. Levels of interferon (IFN)gamma and interleukins IL-12, IL-10, IL-4, IL-5 and IL 13 were evaluated in the culture supernatants with specific immunoassays. RESULTS: Exposure of PBMC to RVs induced the production of IFNgamma, IL-12, IL 10, and IL-13. Cells from asthmatic subjects produced significantly lower levels of IFNgamma and IL-12 and higher levels of IL-10 than normal subjects. IL-4 was induced only in the asthmatic group, while the IFNgamma/IL-4 ratio was more than three times lower in the asthmatic group. CONCLUSIONS: This evidence suggests that the immune response to RVs is not uniquely of a type 1 phenotype, as previously suggested. The type 1 response is defective in atopic asthmatic individuals, with a shift towards a type 2 phenotype in a way similar, but not identical, to their aberrant response to allergens. A defective type 1 immune response to RVs may be implicated in the pathogenesis of virus induced exacerbations of asthma. PMID- 11923552 TI - Home based neuromuscular electrical stimulation as a new rehabilitative strategy for severely disabled patients with chronic obstructive pulmonary disease (COPD). AB - BACKGROUND: Passive training of specific locomotor muscle groups by means of neuromuscular electrical stimulation (NMES) might be better tolerated than whole body exercise in patients with severe chronic obstructive pulmonary disease (COPD). It was hypothesised that this novel strategy would be particularly effective in improving functional impairment and the consequent disability which characterises patients with end stage COPD. METHODS: Fifteen patients with advanced COPD (nine men) were randomly assigned to either a home based 6 week quadriceps femoris NMES training programme (group 1, n=9, FEV(1)=38.0 (9.6)% of predicted) or a 6 week control period before receiving NMES (group 2, n=6, FEV(1)=39.5 (13.3)% of predicted). Knee extensor strength and endurance, whole body exercise capacity, and health related quality of life (Chronic Respiratory Disease Questionnaire, CRDQ) were assessed. RESULTS: All patients were able to complete the NMES training programme successfully, even in the presence of exacerbations (n=4). Training was associated with significant improvements in muscle function, maximal and endurance exercise tolerance, and the dyspnoea domain of the CRDQ (p<0.05). Improvements in muscle performance and exercise capacity after NMES correlated well with a reduction in perception of leg effort corrected for exercise intensity (p<0.01). CONCLUSIONS: For severely disabled COPD patients with incapacitating dyspnoea, short term electrical stimulation of selected lower limb muscles involved in ambulation can improve muscle strength and endurance, whole body exercise tolerance, and breathlessness during activities of daily living. PMID- 11923553 TI - Nationwide prevalence of sporadic and familial idiopathic pulmonary fibrosis: evidence of founder effect among multiplex families in Finland. AB - BACKGROUND: The prevalence of sporadic and familial idiopathic pulmonary fibrosis (IPF) cases in Finland was evaluated according to the revised recommendations of the American Thoracic Society. METHODS: All Finnish pulmonary clinics (n=29) were included in the primary screening. Hospital data bases were used to identify patients with the diagnosis "alveolitis fibroticans idiopathica" (J84.1 in ICD-10 classification). The total number of patients with IPF was extrapolated based on the evaluation of random samples of case records in different centres. Families with more than one potentially affected member were identified from a questionnaire study and the diagnosis was verified from the medical records. RESULTS: Using this approach, the nationwide prevalence of IPF in Finland was estimated to be 16-18/100 000. In 90% of the patients lung involvement was assessed by high resolution computed tomographic (HRCT) scanning and in 31% a surgical biopsy specimen was available, further confirming the diagnosis. Seventeen multiplex families with 2-5 affected family members were identified, giving a prevalence of 5.9/million for familial IPF in Finland. Both multiplex and sporadic families were clustered in Eastern Finland. This clustering reflects the demographic history of Finland in the 16th century and suggests that multiplex families may share a common ancestor in the last 20-25 generations. CONCLUSION: The familial form explained 3.3-3.7% of all Finnish cases of IPF diagnosed according to the revised international guidelines. Geographical clustering of multiplex families suggests a recent founder effect in patients with familial IPF. PMID- 11923554 TI - Randomised trial of intrapleural urokinase in the treatment of childhood empyema. AB - BACKGROUND: The role of intrapleural fibrinolytic agents in the treatment of childhood empyema has not been established. A randomised double blind placebo controlled trial of intrapleural urokinase was performed in children with parapneumonic empyema. METHODS: Sixty children (median age 3.3 years) were recruited from 10 centres and randomised to receive either intrapleural urokinase 40 000 units in 40 ml or saline 12 hourly for 3 days. The primary outcome measure was length of hospital stay after entry to the trial. RESULTS: Treatment with urokinase resulted in a significantly shorter hospital stay (7.4 v 9.5 days; ratio of geometric means 1.28, CI 1.16 to 1.41 p=0.027). A post hoc analysis showed that the use of small percutaneous drains was also associated with shorter hospital stay. Children treated with a combination of urokinase and a small drain had the shortest stay (6.0 days, CI 4.6 to 7.8). CONCLUSION: Intrapleural urokinase is effective in treating empyema in children and significantly shortens hospital stay. PMID- 11923555 TI - Lung restricted T cell receptor AV2S3+ CD4+ T cell expansions in sarcoidosis patients with a shared HLA-DRbeta chain conformation. AB - BACKGROUND: Sarcoidosis is a systemic disease of unknown aetiology frequently affecting the lungs. CD4+ T cells, in particular, accumulate in the lungs, implicating them in the pathogenesis of the disease. METHODS: T cell receptor (TCR) variable (V) gene expression on bronchoalveolar lavage (BAL) fluid T cells and the HLA DR alleles of 121 Scandinavian patients with sarcoidosis was determined. RESULTS: As expected from our previous results, almost every DRB1*0301 (i.e. DR17) positive patient (67/69) had significantly increased numbers of AV2S3+ CD4+ T cells in the BAL fluid but normal levels in peripheral blood (that is, lung restricted expansions) compared with only six of 52 DRB1*0301 negative patients. Detailed genotypic HLA analysis showed that these six DRB1*0301 negative patients with lung restricted AV2S3+ T cell expansions had another HLA allele in common-the HLA-DRB3*0101 allele (also called DR52a)-which was not found in any other DRB1*0301 negative patient. A new group of sarcoidosis patients was therefore identified, characterised by a strict correlation between a distinct HLA allele and lung accumulated T cells expressing a particular TCR V segment. Furthermore, the HLA-DRB1*0301 and HLA-DRB3*0101 encoded molecules showed similarities, with identical amino acid sequences in regions important for antigen binding which may enable them to bind and present the same or similar antigenic peptides. CONCLUSIONS: HLA-DRB3*0101 as well as DRB1*0301 positive sarcoidosis patients may have the capacity to present specific sarcoidosis associated antigens in such a way that AV2S3+ CD4+ T cells are stimulated preferentially, generating lung restricted AV2S3+ T cell expansions. PMID- 11923557 TI - gammadelta T lymphocytes in the peripheral blood of patients with tuberculosis with and without HIV co-infection. AB - BACKGROUND: Several recent studies suggest that gammadelta T lymphocytes play an important role in immunity against Mycobacterium tuberculosis. However, the dynamics of these cells in the peripheral blood of patients with tuberculosis (TB) with and without HIV infection is not fully understood. A study was undertaken to evaluate the profile of the gammadelta T cell population in patients at the time the diagnosis of TB was established. METHODS: A cross sectional study was performed in consecutive TB patients from the Department of Infectious Diseases, Spedali Civili, Brescia. CD4+, CD8+ and Vdelta1 and Vdelta2 T cell counts were analysed. Lymphocyte surface membrane expression was evaluated with the FITC-TCRgammadelta, -Vdelta1, -Vdelta2 and PE-Vdelta1 monoclonal antibodies. Blood donors and HIV seropositive asymptomatic individuals acted as controls. RESULTS: Seventy four TB patients were evaluated, 20 of whom (27%) were co-infected with HIV. HIV seronegative TB patients (n=54) had total gammadelta T cells and Vdelta1 subsets comparable to those in blood donors (n=39). However, the percentage with the Vdelta2 subset was significantly lower in patients with TB than in controls (median 1.5 v 2.1; p=0.05). Responsiveness to PPD was not associated with predominance of a specific gammadelta T cell subset. HIV seropositive individuals had a decreased percentage of circulating Vdelta2 cells at a level similar to that in HIV seronegative TB patients, regardless of the presence of active TB. CONCLUSIONS: HIV seronegative TB patients and HIV infected individuals (with or without active TB) have a reduced number of circulating Vdelta2 T cells compared with healthy individuals. Whether TB and HIV infection share a common mechanism causing Vdelta2 T cell depletion still needs to be established. PMID- 11923556 TI - Expression of p21 in SV40 large T antigen positive human pleural mesothelioma: relationship with survival. AB - BACKGROUND: Mesothelioma is the most commonly occurring primary pleural neoplasm. Several studies have documented an increase in the incidence of this malignancy during the last decades. Although the association between asbestos exposure and development of mesothelioma is generally accepted, the exact mechanism of carcinogenesis is unknown. Recently, Simian virus 40 large T antigen (SV40 Tag) expression has been detected in pleural mesothelioma. The ability of SV40 oncoproteins to inactivate p53 and retinoblastoma tumour suppressor proteins has been proposed as an important step in the pathogenesis of human mesothelioma. METHODS: To obtain a better understanding of the molecular mechanisms of the pathogenesis of mesothelioma, the expression of the cell cycle inhibitor p21(WAF1/CIP1) (p21), a downstream target of p53, was evaluated immunohistochemically in a group of 29 mesothelioma specimens already characterised for the presence of SV40 Tag sequences. RESULTS: Statistical analysis did not reveal any correlation between p21 expression and histopathological type of mesothelioma using the kappa(2) test (p=0.577). A significant positive relationship was found between p21 expression level and the patients' overall survival according to the Kaplan-Meier survival curves and using a log rank test (median difference in survival 7 months, 95% CI 4.8 to 9.9; p<0.001). CONCLUSIONS: Determination of p21 expression bears a prognostic significance in patients affected with mesothelioma, further underlining the role of SV40 in the pathogenesis of malignant pleural mesothelioma. PMID- 11923558 TI - Caffeine decreases exhaled nitric oxide. AB - BACKGROUND: Caffeine is known to inhibit phosphodiesterases, to mobilise intracellular calcium, and to act as an antagonist at adenosine receptors, all of which can potentially alter nitric oxide (NO) production. It was therefore hypothesised that caffeine may alter exhaled NO (eNO) levels. METHODS: In a randomised, single blind, crossover manner, 12 normal subjects consumed either (1) coffee and a placebo capsule, (2) decaffeinated coffee and a capsule of 200 mg caffeine, or (3) decaffeinated coffee and a placebo capsule. Serum caffeine levels were measured at baseline and 1 hour later. Exhaled NO levels were also measured at baseline and each hour for 4 hours. RESULTS: A significant percentage fall in mean (SE) eNO from baseline was seen 1 hour after either caffeinated coffee or a caffeine capsule when compared with placebo (13.5 (4.0)%, p=0.009 and 19.0 (3.8)%, p=0.001, respectively). CONCLUSION: Caffeine causes a significant decrease in eNO which will need to be considered when designing trials to measure eNO levels. The mechanism may be via adenosine receptor antagonism or by altering levels of cGMP. PMID- 11923559 TI - Freeze fracture study of airway epithelium from patients with primary ciliary dyskinesia. AB - BACKGROUND: The airway cilia of patients with primary ciliary dyskinesia (PCD) exhibit several anomalies when studied by transmission electron microscopy, but little is known about the ultrastructural organisation of ciliary membranes in these patients. Freeze fracture replication of airway epithelium from patients with PCD provides a means of achieving high resolution views of cell membrane structure. Ciliary necklaces are a specialised structural feature of ciliary membranes thought to serve as a timing mechanism for ciliary beat, and their characterisation in the cilia of patients with PCD may contribute new insights into the pathophysiology of this syndrome. METHODS: The nasal epithelium of three patients with PCD was freeze fractured and replicated with platinum and carbon shadowing. The resultant preparations were examined by transmission electron microscopy and the ciliary necklaces were compared with similar preparations of nasal biopsy specimens from normal healthy subjects. RESULTS: The ciliary necklaces of the three patients with PCD were normal with no overt differences from those of healthy individuals. CONCLUSIONS: The defective ciliary motility observed in patients with PCD does not appear to result from membrane dysfunction associated with overt disorganisation of ciliary necklace structure. PMID- 11923561 TI - An unusual case of congenital short trachea with very long bronchi mimicking bronchial asthma. AB - Case reports of a short trachea with early branching of the main bronchi are uncommon. The case is presented of a 64 year old woman with upper airway obstruction due to this anatomical abnormality which caused breathlessness and wheezing that was misdiagnosed (and treated) as bronchial asthma for many years. PMID- 11923562 TI - Smoking cessation. PMID- 11923560 TI - The pulmonary physician in critical care * 4: Nosocomial pneumonia. AB - Much progress has been made in the understanding of nosocomial pneumonia but important issues in diagnosis and treatment remain unresolved. The controversy over diagnostic tools should be closed. Instead, every effort should be made to increase our ability to make valid clinical predictions about the presence of ventilator associated pneumonia and to establish criteria to guide restricting empirical antimicrobial treatment without causing patient harm. More emphasis must be put on local infection control measures such as routine surveillance of pathogens, definition of controlled policies of antimicrobial treatment, and effective implementation of strategies of prevention. PMID- 11923563 TI - Conventional RIA underestimates cortisol suppression in the presence of prednisolone. PMID- 11923564 TI - Fibrosing alveolitis in patients with RA. PMID- 11923565 TI - Measuring granulocyte apoptosis in airway inflammation. PMID- 11923566 TI - Aqueous oxygen hyperbaric reperfusion in a porcine model of myocardial infarction. AB - OBJECTIVES: The purpose of the study was to test the hypothesis that intracoronary aqueous oxygen (AO) hyperbaric reperfusion reduces myocardial injury after prolonged coronary occlusion. Background. Attenuation of ischemia/reperfusion injury by the use of hyperbaric oxygen (HBO) administered during reperfusion has been demonstrated for a wide variety of tissues, including myocardium. We have recently developed a more practical, catheter-based, site specific method for delivery of oxygen at hyperbaric levels with aqueous oxygen infusion. METHODS: Following a 60-minute balloon occlusion of the left anterior descending coronary artery in swine, intracoronary AO hyperoxemic perfusion (50 mL blood/minute; 1.5 mL AO/minute; mean pO2 = 834 104 mmHg) was performed for 90 minutes after a 15-minute period of normoxemic autoreperfusion (physiologic reperfusion). Control groups consisted of autoreperfusion alone; active normoxemic perfusion (50 mL/minute) for 90 minutes; and hyperoxemic perfusion with a hollow fiber oxygenator (HFO) for 90 minutes. Results. A significant improvement in left ventricular ejection fraction was noted by ventriculography at 105 minutes of reperfusion (ANOVA, p < 0.05), compared to the 15-minute autoreperfusion period, only in the AO and HFO groups. Mean percent infarct size (area of necrosis)/(area at risk), quantitative post-mortem hemorrhage score, and myocardial myeloperoxidase levels at 3 hours of reperfusion were significantly less in the AO group (ANOVA, p < 0.05), but not in the HFO group, compared to normoxemic groups. Conclusions. The results demonstrate that intracoronary hyperbaric reperfusion with AO, but not with a membrane oxygenator, attenuates myocardial ischemia/reperfusion injury. PMID- 11923568 TI - Incremental progress? The roles of direct stenting and routine troponin measurement after percutaneous coronary intervention. PMID- 11923567 TI - Myocardial injury after apparently successful coronary stenting with or without balloon dilation: direct versus conventional stenting. AB - Direct stenting (i.e., stenting without balloon predilation) is a novel approach to the percutaneous treatment of coronary artery lesions. This approach, by reducing aggression to the vessel wall and immediately sealing the dissections created by balloon inflation by the endoprosthesis, may also significantly lower the rate of procedural ischemic complications. Our purpose was to measure cardiac troponin T (cTnT), creatine kinase and its isoform CKMB after apparently successful elective stent implantation with conventional stenting (CS) or direct stenting (DS) and to compare the procedural myocardial injury between these 2 approaches. Enzyme levels were measured before and 16 hours post-procedure. A second-generation commercial ELISA cTnT assay (Boehringer Mannheim Corporation) was used to measure cTnT with a cut-off of 0.1 ng/dl. There was no abnormality in any of the 3 enzymes in either group before the procedure. Although the incidence of cTnT was elevated in 4 of 37 patients (10.8%) in the DS group and in 5 out of 23 patients (21.7%) in the CS group at 16 hours post-procedure, these values did not reach statistical significance (p > 0.05). Creatine kinase and CKMB levels were not elevated in any of the patients. CTnT and CKMB measurements are needed to detect this minor myocardial damage. Randomized studies with larger patient populations should be conducted to compare the two different approaches. PMID- 11923569 TI - Safety and efficacy of using 0.052-inch Gianturco coil for closure of large ( > or = 4mm) patent ductus arteriosus. AB - Transcatheter closure of patent ductus arteriosus (PDA) is now a well-established treatment alternative to surgery in many cardiology centers. Of all the methods used, transcatheter coil occlusion is the preferred therapy. For small PDA, the method using 0.038" Gianturco coils has proven safe and effective. However, this therapeutic strategy has encountered some difficulties with large PDA. This study provides an alternative strategy, using 0.052" Gianturco coil and complete closure of residual shunt with multiple coils to close large PDA. Fifteen patients underwent transcatheter coil occlusion of large ( > or = 4mm) patent ductus arteriosus. The intermediate success rate was 86.7%. There were four complications and only two patients had to be referred for surgery. PMID- 11923570 TI - Coil closure of the large patent arterial duct: cheap, but how easy and effective? PMID- 11923571 TI - Laser angioplasty and laser-induced thrombolysis in revascularization of anomalous coronary arteries. AB - Acute coronary syndromes such as unstable angina and myocardial infarction are attributed to a pathophysiologic process that involves rupture of atherosclerotic plaque and subsequent thrombosis. Percutaneous intervention of anomalous coronary arteries in patients who present with acute coronary syndromes impose unique technical challenges related to the specific anatomic course and morphology of these vessels. Selection of appropriate guiding catheter configuration, choice of supportive guidewire, and proper delivery and activation of debulking devices and stents are important steps toward achieving adequate results. Excimer laser angioplasty is a debulking technology for removal of atherosclerotic plaque and associated thrombi. To date, application of laser angioplasty in anomalous coronary arteries is unreported. We herein present clinical data and discuss technical aspects related to performance of excimer laser angioplasty in three symptomatic patients with acute coronary syndrome, two having an anomalous right coronary artery and one with an anomalous circumflex artery. The delivery of laser energy in these cases resulted in rapid thrombolysis of an occlusive thrombus, successful debulking of the underlying atherosclerotic plaque, facilitation of adjunct balloon angioplasty and stenting, and ultimately, improved clinical condition. PMID- 11923572 TI - Predictors of recurrent restenosis after coronary stenting: an analysis of 197 patients. AB - One of the major limitations in coronary stenting is in-stent restenosis. This study was aimed to identify clinical, angiographic, and procedural factors that may be related to recurrent in-stent restenosis. We analyzed consecutive 197 patients who underwent coronary stenting. Follow-up angiography was available in 170 patients and repeat balloon angioplasty was performed for in-stent restenosis. These patients were subdivided into 3 groups: group A consisted of 100 patients that were never restenosed, group B had 49 patients restenosed once, and in group C were 21 patients restenosed more than twice. Group C was more often female (48%) and included diabetes mellitus patients (52%). Lesion location, reference vessel size and diameter stenosis were similar for all groups. However, the incidence of calcified lesions tended to be higher (50% vs. 29%; p = 0.07), and lesion length was longer in group C than in group A (11.9+/- 5.4 mm vs. 9.0+/- 3.9 mm; p < 0.01). Diameter stenosis after predilation as well as after stenting was significantly higher in group C than in group A (50+/- 10% vs 39+/- 10%; p < 0.01, 32+/- 8% vs. 19+/- 10%; p < 0.01). The incidence of diffuse type of in-stent restenosis was significantly higher in group C than in group B (62% vs. 14%; p < 0.01). Multivariate logistic regression analysis identified diameter stenosis after stenting (p = 0.0022), female (p = 0.0135), and diameter stenosis after predilatation (p = 0.0233) as the significant correlate of recurrent in-stent restenosis. In conclusion, the major recurrent in stent restenosis predictors identified included female gender, final diameter stenosis, and diameter stenosis after predilatation. PMID- 11923573 TI - Implantation of a dual-chamber pacemaker in a patient with persistent left superior vena cava using a a steerable stylet. AB - A persistent left superior vena cava (PLSVC) is a rare abnormality that presents a challenge to the implanter. To our knowledge, this is the first case report of implantation of dual-chamber pacemaker via PLSVC utilizing a steerable stylet (the Locater; St. Jude Medical, Inc., Minnetonka, Minnesota) with chronic follow up showing stable threshold. PMID- 11923574 TI - Primary stenting in a patient with acute myocardial infarction and primary antiphospholipid syndrome. AB - We present a patient who had recurrent coronary thrombosis due to primary antiphospholipid syndrome. The patient was treated with streptokinase during the first myocardial infarction and primary stenting during reinfarction. To our knowledge, this is the first case in which the occluded right coronary artery was treated with primary stent implantation. PMID- 11923575 TI - Spontaneous coronary artery dissection in a young woman precipitated by retching. AB - Spontaneous coronary artery dissection as a cause of acute myocardial ischemia is a rare entity that has been associated with several different clinical profiles and precipitating events. The recognition of this entity as the cause of acute ischemia is important because the therapeutic considerations may be different than that for ischemia due to a ruptured atherosclerotic plaque. We report a case of spontaneous coronary artery dissection in a 31-year-old female that was induced by prolonged, forceful retching. To our knowledge, this is the first reported case of such an association. Prolonged retching should be added to the list of causes of spontaneous coronary artery dissection. PMID- 11923576 TI - Late-term myocardial infarction after surgical ligation of a giant coronary artery fistula. AB - A case is presented of a patient with recurrent myocardial infarctions after surgical treatment of a giant coronary artery fistula. The etiology was due to thrombus development in a large blind pouch, with propagation into more proximal vessels. Stenting of the proximal vessel provided temporary benefit, but recurrence eventually required surgical closure of the blind pouch. PMID- 11923577 TI - Spontaneous dissection bilateral internal mammary arterial grafts. AB - Spontaneous coronary artery dissection is rare. We report a case of a 49-year-old male with quadruple bypass grafts who presented with acute myocardial infarction due to spontaneous dissection of both internal mammary artery grafts. PMID- 11923578 TI - Extensive right coronary artery dissection following cutting balloon treatment of in-stent restenosis. PMID- 11923579 TI - Iliac artery stenosis causing post-renal transplant hypertension: successful management by percutaneous angioplasty and stent implantation. AB - Atherosclerotic occlusion of the native iliac arteries and/or transplant renal artery is a major cause of post-transplant hypertension. Iliac artery stenosis mimics renovascular hypertension and may cause renal dysfunction in transplant recipients. We report a case of a 61-year-old renal transplant recipient with native bilateral iliac artery stenoses and coronary artery disease. He presented with severe hypertension and was managed successfully with angioplasty and stenting of native iliac arteries. PMID- 11923580 TI - Risk stratification in patients with unstable angina and non-ST segment elevation myocardial infarction: evidence-based review. PMID- 11923581 TI - Ethanol decreases the efficiency of phosphorylation of thymidine kinase in a human T-lymphocytic cell line. AB - BACKGROUND: Thymidine kinase (TK) and thymidylate kinase (TMPK) are the two rate limiting enzymes in the cascade of activation of the anti-human immunodeficiency virus (HIV) drug 3'-azido-3'-deoxythymidine (AZT) to its active triphosphate form. We examined the effect of ethanol and a combination of ethanol and AZT on TK and TMPK activities in human Jurkat T cells. METHODS: Jurkat T cells were exposed to 0.2 and 0.5% (v/v) ethanol concentrations alone or in combination with 5 or 10 microM AZT for 48 hr in growth medium. TK and TMPK activities were determined by measuring the conversion of [3H] substrates (thymidine or AZT for TK and thymidine monophosphate for TMPK) to their respective monophosphate or diphosphate forms. The effect of ethanol on the transcriptional activity of TK was determined by reverse transcription-polymerase chain reaction and on the growth of Jurkat cells by [3H]-thymidine incorporation and cell-cycle analysis. RESULTS: Treatment of Jurkat cells with 0.2 and 0.5% ethanol concentrations resulted in 25 and 50% decreases (p < or = 0.05) in TK activity, respectively. No significant changes were observed in the TMPK activities. However, ethanol decreased the formation of thymidine diphosphate from thymidine in coupled TK/TMPK reactions, suggesting that decreases in TK activity could result in an overall decrease in the phosphorylation of AZT. The effect of ethanol on TK was independent of its transcript level. AZT in combination with ethanol decreased the inhibitory effect of ethanol on TK activity. However, it did not block the ethanol effect even at higher concentrations. Ethanol significantly decreased the proliferative capacity and cell cycle progression of the Jurkat cells. CONCLUSIONS: Our in vitro study in human Jurkat T cells indicates that at physiologically achievable concentrations in humans, ethanol can decrease TK activity through decreases in cell proliferation, and it suggests that ethanol ingestion in HIV-1-infected individuals could compromise activation of AZT and related drugs through decreased TK activity. PMID- 11923582 TI - Alcohol-related ERP changes recorded from different modalities: a topographic analysis. AB - BACKGROUND: There is controversy in the literature regarding the relationship between event-related-potential (ERP) abnormalities in abstinent alcoholics and stimulus-processing modality (i.e., visual versus auditory). The first purpose of this study was to address questions about whether ERP abnormalities observed in alcoholics are modality specific. The second purpose was to employ current source density (CSD) analyses to investigate topographic differences between alcoholics and controls within each modality. METHODS: Data were collected from 30 sober male alcoholics and 39 normal males in a typical auditory oddball task and in a visual oddball paradigm with novel stimuli, with an extensive set of 61 scalp electrodes. Visual and quantitative assessment of CSD maps as well as analyses of variances on both raw and normalized ERP data were performed. RESULTS: Positive findings were limited to the N1 and P3 components. The visual N1 amplitude was significantly smaller in alcoholics than in controls at the parietal region; no significant group differences in N1 were found in the auditory modality. Alcoholics had widespread reductions in P3 amplitudes in both modalities compared with controls, although in the frontal region this effect was partially due to the influence of age. These P3 reductions in alcoholics were statistically more pronounced in the posterior compared with the anterior regions regardless of modality. Topographically, sources in CSD maps were weaker in alcoholics than in controls; in the frontal and central regions, the weakness was more pronounced in the auditory modality but, in parietal and occipital regions, it was more pronounced in the visual modality. CONCLUSIONS: The results suggest that, in abstinent alcoholics, abnormalities in auditory ERPs may be localized to more anterior sources, while abnormalities in visual ERPs may be localized to more posterior sources. ERP topographic features are more sensitive than amplitude measurements in assessing alcoholic-related modality effects. PMID- 11923583 TI - Microdialysis of dopamine in the nucleus accumbens of alcohol-preferring (P) rats during anticipation and operant self-administration of ethanol. AB - BACKGROUND: The present study was designed to test directly whether a contextual stimulus for access to ethanol would acquire the ability to enhance locomotor activity and dopamine efflux in the nucleus accumbens (NAc) of alcohol-preferring (P) rats. The study also explored the association between elevated locomotor activity and NAc dopamine efflux during operant self-administration of ethanol. METHODS: Adult female P rats were randomly assigned to operantly self-administer either 15% (v/v) ethanol or 0.0125% (w/v) saccharin. Both groups were trained in a daily 30-min two-lever concurrent operant task (FR-3) to orally self-administer ethanol or saccharin, with water on the alternate lever. A third (control) group was trained to self-administer water on both levers. All groups were also acclimated in the operant chambers to periods of habituation, anticipation, and postadministration. RESULTS: Compared with controls, the ethanol group, but not the saccharin group, showed significant increases in locomotor activity as well as increased NAc dopamine efflux during the first 10 min of the anticipation period. During the first 10 min of the self-administration period, locomotor activity was significantly increased in both the ethanol and saccharin groups compared with control values. The ethanol group, but not the saccharin group, showed significant increases in NAc dopamine efflux during the 20th and 30th min of the self-administration period and during the first 10 min of the postadministration period. CONCLUSIONS: The findings suggest that acquisition of signal-induced anticipation of self-administered ethanol is associated with increases in locomotor activity and extracellular levels of dopamine in the NAc of P rats. Such associations may be important to the development and maintenance of ethanol-seeking behaviors. The findings also indicate that operant self administration of ethanol is associated with increases in extracellular levels of dopamine in the NAc of P rats. PMID- 11923584 TI - Mecamylamine modifies the pharmacokinetics and reinforcing effects of alcohol. AB - BACKGROUND: Central nicotinic cholinergic receptors modify alcohol-induced mesolimbic dopamine activation, which seems to be important in the reinforcing properties of alcohol. Consistent with this model, acute administration to rats of the tertiary nicotinic receptor antagonist mecamylamine blocks both alcohol consumption and alcohol-induced dopamine release in the nucleus accumbens. This study was conducted to test the hypothesis that, during the ascending limb of the blood alcohol concentration curve, mecamylamine would reduce the stimulating and pleasurable effects of an intoxicating dose of alcohol in humans. METHODS: Ten female and 10 male volunteers with no history of alcohol or substance use disorders, including nicotine dependence, completed the study. During two laboratory sessions, subjects consumed three aliquots of an alcohol-containing drink, with a total ethanol content of 0.7 g/kg (in women) or 0.8 g/kg (in men), over a 30-min period. Two hours before the first drink, subjects were pretreated with mecamylamine or placebo, with the order of sessions counterbalanced. Primary outcome measures included the Drug Effect Questionnaire, the central stimulation subscale of the Alcohol Sensation Scale, and the stimulant subscale of the Biphasic Alcohol Effects Scale. Breath alcohol level (BAL) was examined to identify the ascending and descending limbs of the blood alcohol curve and to assess pharmacokinetic interactions between alcohol and mecamylamine. RESULTS: Significant effects of time, study drug, and their interaction were observed. Compared with placebo, mecamylamine reduced BAL. After controlling for BAL at each time point, mecamylamine also reduced the Drug Effect Questionnaire and Alcohol Sensation Scale stimulant subscale scores, with a trend for a similar effect on the Biphasic Alcohol Effects Scale score. CONCLUSIONS: Mecamylamine seems to modify both the pharmacokinetic profile of alcohol and the rewarding effects of alcohol in healthy volunteers. PMID- 11923585 TI - CDT, GGT, and AST as markers of alcohol use: the WHO/ISBRA collaborative project. AB - BACKGROUND: Estimates of the performance of carbohydrate deficient transferrin (CDT) and gamma glutamyltransferase (GGT) as markers of alcohol consumption have varied widely. Studies have differed in design and subject characteristics. The WHO/ISBRA Collaborative Study allows assessment and comparison of CDT, GGT, and aspartate aminotransferase (AST) as markers of drinking in a large, well characterized, multicenter sample. METHODS: A total of 1863 subjects were recruited from five countries (Australia, Brazil, Canada, Finland, and Japan). Recruitment was stratified by alcohol use, age, and sex. Demographic characteristics, alcohol consumption, and presence of ICD-10 dependence were recorded using an interview schedule based on the AUDADIS. CDT was assayed using CDTect and GGT and AST by standard methods. Statistical techniques included receiver operating characteristic (ROC) analysis. Multiple regression was used to measure the impact of factors other than alcohol on test performance. RESULTS: CDT and GGT had comparable performance on ROC analysis, with AST performing slightly less well. CDT was a slightly but significantly better marker of high risk consumption in men. All were more effective for detection of high-risk rather than intermediate-risk drinking. CDT and GGT levels were influenced by body mass index, sex, age, and smoking status. CONCLUSIONS: CDT was little better than GGT in detecting high- or intermediate-risk alcohol consumption in this large, multicenter, predominantly community-based sample. As the two tests are relatively independent of each other, their combination is likely to provide better performance than either test alone. Test interpretation should take account sex, age, and body mass index. PMID- 11923586 TI - Metadoxine in acute alcohol intoxication: a double-blind, randomized, placebo controlled study. AB - BACKGROUND: At present there are only intriguing and preliminary clinical results regarding the efficacy of metadoxine (pyridoxol L-2-pyrrolidone-5-carboxylate) in acute alcohol intoxication. The present study was planned with the aim of investigating the effectiveness of metadoxine in the management of patients affected by acute ethanol intoxication. METHODS: A double-blind, randomized, multicenter, placebo-controlled trial was carried out on 58 patients of both sexes with acute ethanol intoxication. Patients were treated with a single dose of 900-mg intravenous metadoxine (n = 29) or with placebo (n = 29). Patients were clinically and biochemically evaluated at 0.5, 1, 2, 3, 6, 9, and 12 hr after treatment. RESULTS: Treatment with metadoxine significantly decreased the half life of ethanol in blood (from 6.70 +/- 1.84 to 5.41 +/- 1.99 hr; p < 0.013) and showed a faster rate of ethanol elimination. The effects on ethanol half-life in blood were accompanied by a faster onset of recovery from intoxication, defined as the time of the transition of blood ethanol levels to the immediately lower range defined by intoxication categories (in g/liter: 0 to 0.5, absent; 0.51 to 1.0, mild; 1.1 to 2.5, moderate; >2.5, severe). Thus the median time to onset of recovery was 0.95 hr with metadoxine and 2.34 hr with placebo (p = 0.013). The effects of treatment on blood alcohol levels were paralleled by a significant decrease in the rating of the toxic clinical symptomatology. At 2 hr the improvement of toxic symptoms (in percent of maximum possible) was 68 +/- 28 vs. 44 +/- 27% in controls (p < 0.002). CONCLUSIONS: In patients with acute ethanol intoxication metadoxine accelerated the elimination of ethanol from blood, which led to faster recovery from intoxication, and improved the behavioral toxic symptomatology. Metadoxine could be helpful in the management of acute ethanol intoxication. PMID- 11923587 TI - Paternal alcohol exposure affects sperm cytosine methyltransferase messenger RNA levels. AB - BACKGROUND: Although paternal alcohol exposure has been shown to affect the growth and behavior of offspring, the mechanisms underlying these effects still remain to be elucidated. This study examines one possible mechanism, namely, altered genomic imprinting as reflected by changes in sperm cytosine methyltransferase messenger RNA (mRNA) levels. METHODS: Male rats were treated with alcohol for 9 weeks before breeding. Resulting fetuses were counted and weighed, and paternal sperm was examined for changes in cytosine methyltransferase mRNA levels. RESULTS: Alcohol did not affect mating, fecundity, or litter size, but it did result in significantly decreased mean fetal weight, increased fetal runt incidence in offspring, and decreased cytosine methyltransferase mRNA levels in paternal sperm, compared with pair-fed and ad libitum controls. CONCLUSIONS: Alcohol-induced reductions in cytosine methyltransferase mRNA levels may reflect altered genomic imprinting caused by reduced DNA methylation, which, in turn, may lead to the expression of normally silent paternal alleles and may be a mechanism for paternal alcohol effects. PMID- 11923588 TI - Grape polyphenols inhibit chronic ethanol-induced COX-2 mRNA expression in rat brain. AB - BACKGROUND: Chronic ethanol has been shown to increase oxidative stress leading to neurodegenerative changes in the brain. Oxidative stress may up-regulate extracellular signal regulated kinases (ERK1/2) and, subsequently, the arachidonic acid cascade mediated by phospholipase A2 (PLA2) and cyclooxygenase (COX-2). Our earlier study showed that grape polyphenols (GP) could ameliorate oxidative damage to synaptic membrane proteins due to chronic ethanol treatment. This study was aimed at examining the effects of GP on mRNA expression of ERK1/2, cytosolic PLA2 (cPLA2), and COX-2 in different brain regions after chronic ethanol treatment. METHODS: Male Sprague-Dawley rats were fed a Lieber-DeCarli liquid diet with ethanol or isocaloric amount of maltose, with or without GP for 2 months. In situ hybridization was carried out using coronal brain sections through the hippocampus. RESULTS: Quantitative in situ hybridization showed no changes in ERK1 and cPLA2 mRNA levels in cortical areas and hippocampus after ethanol and/or GP administration. However, a decrease in ERK2 and an increase in COX-2 mRNA level was found in the hippocampus of ethanol-treated animals. GP completely inhibited the increase in COX-2 due to ethanol treatment. CONCLUSION: Increase in COX-2 expression may be an underlying mechanism for the increase in oxidative stress induced by chronic ethanol administration. Dietary supplementation of GP may have a beneficial role in inhibiting certain alcohol effects. PMID- 11923589 TI - Inhibitory effects of ethanol on rat mesangial cell proliferation via protein kinase C pathway. AB - A large body of evidence has shown that ethanol inhibits the cell growth and cell proliferation in a variety of cell types. However, it has not been studied whether ethanol inhibits the proliferation of mesangial cells (MC) in the kidney. We examined the effects of ethanol on cell proliferation in cultured rat MC. Treatment with ethanol (10-200 mM) for 48 hr inhibited [(3)H]thymidine incorporation into MC in a concentration-dependent manner. The same concentrations of ethanol also inhibited the increase in cell number of MC. GF109203X and chelerythrine chloride, inhibitors for protein kinase C, eliminated the inhibitory effects of ethanol; and protein kinase C activator, PMA, mimicked the effects of ethanol. In contrast, neither the protein kinase A inhibitor H-89 nor the protein kinase G inhibitor KT5823 had any effect. These findings suggest that ethanol has inhibitory effects on the proliferation of MC, probably via activation of the protein kinase C pathway. PMID- 11923590 TI - Ethanol enhances activation-induced caspase-3 dependent cell death in T lymphocytes. AB - BACKGROUND: Clinical and experimental studies have shown that an important deleterious consequence of excessive alcohol consumption is immunosuppression, specifically, a depletion in the mature CD4+ T-cell population. A predominant mechanism involved in T-cell depletion is activation-induced cell death (AICD). Although it is well documented that ethanol intake can cause depletion of CD4+ T cells, the mechanism of how alcohol mediates its effects is unclear. METHODS: The results were based on data from three separate experiments presented as mean +/- standard deviation (SD). Jurkat CD4+ T cells and peripheral blood lymphocytes were treated with 25 mM of ethanol (12-18 hr), followed by stimulation with mitogens Conconavalin A (5 microg/ml) and Phytohemmaglutinin (1 microg/ml) or T cell receptor ligation (anti-CD3 antibody (5 microg/ml)) for 6 hr, and then harvested for measurement. The apoptotic cell death markers measured include cell viability, Caspase-3-like activity, and DNA fragmentation. RESULTS: We demonstrate that alcohol pretreatment enhances AICD of Jurkat CD4+ T cells and peripheral blood lymphocytes upon activation by CD3-crosslinking or stimulation with Conconavalin A and Phytohemmaglutinin. Furthermore, we find that the ethanol mediated enhancement of T cells to apoptosis involves increased activation of Caspase-3 and can be abrogated by treatment with a specific inhibitor of Caspase 3. CONCLUSIONS: Our data indicate that ethanol can sensitize CD4+ T cells to enhanced stimulation-induced Caspase-3 activation and to subsequent AICD. This is, perhaps, an important mechanism in alcohol-induced immunosuppression. PMID- 11923591 TI - Effects of lorazepam treatment for multiple ethanol withdrawals in mice. AB - BACKGROUND: Although many alcohol-dependent patients present with a history of prior detoxifications, the efficacy and safety of pharmacotherapy in the context of multiple ethanol withdrawal experiences have not been extensively studied. The purpose of this study was to evaluate the ability of lorazepam treatment for multiple withdrawals to prevent or blunt the development/expression of sensitized central nervous system hyperexcitability during a subsequent untreated withdrawal episode. A mouse model of withdrawal sensitization involving repeated ethanol withdrawals was used. METHODS: Adult male C3H/He mice were exposed to different patterns of chronic ethanol vapor in inhalation chambers. One group received four cycles of 16 hr of ethanol exposure separated by 8-hr withdrawal periods, another group was tested after a single 16-hr exposure period, and a final group served as ethanol-naive controls. These groups were further divided into lorazepam dosage (0.25-1.0 mg/kg) conditions. Lorazepam was administered 1 hr into each of the first three withdrawal cycles (or equivalent times); no drug injections were given during the final (fourth) withdrawal cycle. The ability of lorazepam treatment to alter development and expression of sensitized handling-induced convulsions (HIC), as well as changes in pentylenetetrazol seizure threshold dosage during an untreated withdrawal episode, was examined. Separate animals were used to assess the effects of lorazepam treatment on blood ethanol clearance and plasma levels of the benzodiazepine during the test withdrawal cycle. RESULTS: Lorazepam dose-dependently reduced HIC activity during successive withdrawal cycles, and this resulted in attenuated expression of the sensitized HIC response during the acute phase of a subsequent untreated withdrawal episode. However, HIC activity was exacerbated at later time points during this final test withdrawal in mice that had received lorazepam treatment for earlier withdrawals. A similar pattern of results was obtained for changes in pentylenetetrazol seizure threshold dosage. These results do not seem to be due to pharmacokinetic factors, because peak blood ethanol levels, rate of ethanol elimination, and plasma levels of lorazepam did not significantly differ among groups during the final test withdrawal cycle. CONCLUSIONS: Blocking central nervous system hyperexcitability during repeated ethanol withdrawals with lorazepam effectively blunts the development and expression of sensitized seizure activity during the acute phase of a subsequent unmedicated withdrawal episode. At later time points, withdrawal-related seizure activity was exacerbated, and this is possibly reflective of an interaction between protracted ethanol withdrawal and withdrawal from the benzodiazepine. The clinical implications of these findings suggest that repeated use of benzodiazepines for treatment of multiple ethanol withdrawals may have some initial beneficial effects, but such treatment may also place patients at increased risk of seizures at later time points. PMID- 11923592 TI - Ethanol-enhanced cytotoxicity of alkylating agents. AB - BACKGROUND: Although ethanol itself is not genotoxic, chronic alcohol consumption increases the risk of neoplastic disease. The mechanism by which ethanol exerts a cocarcinogenic effect is not well established, and the aim of this study was to determine whether exposure to ethanol increased the cytotoxicity of known carcinogens. METHODS: To assess cell survival, the ability of Chinese hamster A10 cells, which express alcohol dehydrogenase, to form colonies was determined after exposure to ethanol and other substances, including both genotoxicants and non DNA-reactive cytotoxic agents. RESULTS: 1-Methyl-3-nitro-1-nitrosoguanidine (MNNG) is an alkylating agent that forms covalent bonds with DNA. The cytotoxicity of MNNG at concentrations of 0.17 to 0.68 microM was markedly enhanced when cells were also treated with 50 mM ethanol. When combined with 0.34 microM MNNG, concentrations of ethanol as low as 2 mM exacerbated the toxicity of this alkylating agent. When these experiments were repeated in the presence of 4 methylpyrazole, an inhibitor of alcohol dehydrogenase, pretreatment with ethanol did not affect the toxicity of MNNG. When ethanol treatment was combined with exposure to other carcinogens, as well as agents that do not directly damage DNA, the cytotoxicity of the DNA-reactive agents 4-nitroquinoline-N-oxide, mitomycin C, and 6-chloro-9-(3-[2-chloroethyl]aminopropylamino)-2-methoxyacridine was modestly enhanced, and that of a second alkylating agent, ethyl methanesulfonate, was markedly increased. CONCLUSIONS: The results are consistent with impairment of DNA repair processes, particularly base excision repair, by acetaldehyde, as a mechanism by which ethanol increases the genotoxicity of certain genotoxic agents. PMID- 11923593 TI - Chronic ethanol treatment and withdrawal alter ACPD-evoked calcium signals in developing Purkinje neurons. AB - BACKGROUND: Alcohol exposure during human fetal development can result in fetal alcohol syndrome, a condition characterized by central nervous system dysfunction. Detailed studies in animal models of fetal alcohol syndrome show that the cerebellar region is particularly sensitive to alcohol exposure during early development; however, the cellular mechanisms underlying the alcohol sensitivity of the immature cerebellum are poorly understood. METHODS: Primary neuronal cultures of cerebellar cells were prepared from embryonic day 20 rat pups. Cultures were exposed to ethanol (33 mM; 150 mg/100 ml) during the main period of morphological development of the Purkinje neurons, from 6 to 17 days in vitro. After the ethanol treatment, the response of Purkinje neurons to the selective metabotropic glutamate receptor (mGluR) agonist (1S,3R)-1 aminocyclopentane-1,3-dicarboxylic acid (ACPD; 300 microM) was examined in parallel fura-2 Ca2+ imaging and current-clamp experiments. In an additional set of experiments, ethanol-treated cultures were allowed to withdraw from ethanol overnight before recordings were performed. RESULTS: In Ca2+ imaging studies, the mean peak amplitude of ACPD-evoked Ca2+ signals was depressed in the dendritic region of chronic ethanol-treated Purkinje neurons compared with control neurons (p < 0.05, unpaired t test), whereas there was no apparent difference in the somatic region. In contrast, peak ACPD-evoked Ca2+ signals were enhanced in both the somatic and dendritic regions of withdrawn Purkinje neurons compared with control neurons. Parallel current-clamp studies showed no consistent effect of chronic ethanol treatment or ethanol withdrawal on the membrane response to ACPD. CONCLUSIONS: These results show that prolonged ethanol exposure and early withdrawal lead to alterations in mGluR-evoked Ca2+ signaling in cerebellar Purkinje neurons. Metabotropic GluRs in the Purkinje neuron play important roles in cerebellar development and function, suggesting that alterations of mGluR signaling pathways by ethanol may play a key role in the actions of ethanol on the developing cerebellum. PMID- 11923594 TI - Effects of combined systemic alcohol and central nicotine administration into ventral tegmental area on dopamine release in the nucleus accumbens. AB - BACKGROUND: Alcoholism is associated with a higher incidence of smoking. The mesolimbic dopaminergic pathway is believed to play an important role in the reinforcing effects of both ethanol and nicotine. This study was undertaken to determine whether simultaneous administration of systemic ethanol and microinjection of nicotine into the ventral tegmental area (VTA) would result in exaggerated release of dopamine (DA) in the shell region of nucleus accumbens. METHODS: Microdialysis was applied in awake, freely moving adult male Wistar rats, and DA concentration in the dialysate was measured by HPLC-electrochemical detectors. RESULTS: Systemic administration of ethanol or microinjection of nicotine into VTA resulted in a dose-dependent increase in DA release (extracellular DA concentration). Simultaneous administration of lower doses of nicotine and ethanol resulted in an additive effect on the released DA. This additive effect was not observed with higher doses of nicotine and ethanol. Administration of the nicotinic antagonist mecamylamine into VTA completely blocked ethanol-induced DA release. CONCLUSIONS: These data support the hypothesis that the reinforcing effects of ethanol are at least partially mediated through the nicotinic receptors in the VTA. Furthermore, administration of selective nicotinic antagonists may be of therapeutic potential in reducing the rewarding effects of ethanol. The data also suggest that the combined effects of ethanol and nicotine on the "reward pathway" may be a contributing factor to the high incidence of smoking in alcoholics. PMID- 11923595 TI - Corpus callosum, pons, and cortical white matter in alcoholic women. AB - BACKGROUND: To measure the effect of alcohol abuse on white matter brain macrostructure in women with alcoholism and to determine whether observed abnormalities interact with age. METHODS: Quantitative measures of corpus callosum area, cortical white matter volume, and pons volume were derived from magnetic resonance imaging scans obtained from 34 women with DSM-III-R alcoholism (aged 28-64, mean 41 years) and 35 healthy women (aged 22-65, mean 42 years). Transverse relaxation time of the pons was also obtained. RESULTS: No significant group differences in any brain measures were observed. However, in alcoholics greater length of sobriety was associated with more cortical white matter, and higher lifetime levels of alcohol consumption were associated with smaller volumes and prolonged transverse relaxation time in the pons. CONCLUSIONS: Despite a lack of overall deficits in white matter macrostructural size in alcoholic women, certain white matter structures showed alcohol exposure vulnerability whereas others showed evidence of recovery with abstinence. PMID- 11923597 TI - Effects of managed care on alcohol and other drug (AOD) treatment. AB - The article represents the proceedings of a symposium at the 2001 RSA Meeting in Montreal, Canada. The organizer/chair was Stephen Magura. The presentations examined: (1) How managed care organization policies may affect enrollees' use of alcohol and other drug (AOD) treatment, by Constance Horgan and associates; (2) The determinants of patients' access to and utilization of AOD treatment in a large health maintenance organization, by Jennifer R. Mertens and Constance Weisner; (3) The impact on treatment access and costs of a statewide carve-out for AOD treatment for Medicaid, by Donald Shepard and associates; and (4) The predictive validity of a new patient assessment technology developed, in part, to better justify AOD treatment in response to the demands of managed care, by Stephen Magura and associates. PMID- 11923596 TI - Ethanol-induced translocation of protein kinase A occurs in two phases: control by different molecular mechanisms. AB - BACKGROUND: Cyclic adenosine monophosphate (cAMP)-dependent protein kinase A (PKA) regulates cellular functions. The specificity of PKA-mediated phosphorylation is determined primarily by PKA localization to sub-cellular sites. Chronic exposure to ethanol causes sustained translocation of the PKA catalytic subunit (C) from the Golgi to the nucleus in NG108-15 cells. Here we find that this is preceded by a transient short-term ethanol-induced translocation of PKA C. Different molecular mechanisms appear to underlie early and late phases of ethanol-induced translocation of PKA subunits. METHODS: The time course and localization of PKA C and regulatory (RII) subunits was assessed by immunocytochemistry in NG108-15 cells in the presence of ethanol, adenosine receptor (A2) blockade, and inhibitors of PKA activity and RNA and protein synthesis. RESULTS: Ethanol induces an early phase (<30 min) of C translocation to the cytoplasm and nucleus. This requires cAMP via adenosine A2 receptor activation. C then returns to the Golgi area after 60 min. A second phase of C translocation occurs during continuing exposure to ethanol (>12 hr). Re accumulation of nuclear C no longer requires A2 or cAMP. RII also translocates to the nucleus during chronic treatment with ethanol. Both C and RII remain in the nucleus as long as ethanol is present. Unlike the early phase of ethanol induced translocation, the second phase of PKA subunit translocation requires protein and RNA synthesis. CONCLUSIONS: We identify two distinct phases of ethanol-induced PKA translocation which appear to be regulated by different molecular mechanisms. The first requires A2 signaling and cAMP; the later phase requires RNA and protein synthesis. The two phases of ethanol-induced PKA translocation observed in cell lines may contribute to changes in PKA signaling, cAMP-dependent gene expression, and the initiation and maintenance of sustained drinking behavior in experimental animals. PMID- 11923598 TI - Using acquired knowledge and new technologies in alcoholism treatment trials. AB - This article represents the proceedings of a symposium held at the 2001 RSA meeting in Montreal, Canada. The organizer and chair was Barbara A. Flannery and the discussant was Raye Z. Litten. The presentations were (1) The use of biomarkers in alcohol-dependence treatment trials, by John P. Allen; (2) Strategies for enhancing patient compliance in clinical treatment trials, by Helen M. Pettinati; (3) The predictive utility of an alcohol-craving measure, by Barbara A. Flannery; (4) What should be the primary outcome measures in a clinical trial, by Damaris J. Rohsenow; (5) Innovative strategies for assessing functional outcomes in alcoholism treatment clinical trials, by Ron A. Cisler. PMID- 11923599 TI - Rates of circumferential resection margin involvement vary between surgeons and predict outcomes in rectal cancer surgery. AB - OBJECTIVE: To analyze the potential variability in rates of circumferential resection margin (CRM) involvement between different surgeons and time periods and to determine the suitability of using CRM status as an immediate predictor of outcome after rectal cancer surgery. SUMMARY BACKGROUND DATA: After disease stage has been taken into account, survival in rectal cancer has been shown to be very variable between surgeons and institutions. One of the major factors influencing survival is local recurrence, and this in turn is strongly related to inadequate tumor excision, particularly at the CRM. METHODS: In a study involving 608 patients who underwent surgery for rectal cancer in Leeds during the 12-year period 1986 to 1997, the authors examined the role of CRM status as an immediate predictor of likely outcome, paying particular attention to its relationships with different surgeons and time periods. RESULTS: Of 586 patients on whom full clinical follow-up was obtained, 165 (28.2%) had CRM involvement by carcinoma on pathologic examination. Up to the end of 1998, 105 (17.9%) patients had developed local recurrence. A significantly higher proportion (38.2%) of CRM-positive patients developed local recurrence than CRM-negative ones (10.0%). Kaplan-Meier survival analysis showed significant improvements in survival for CRM-negative patients over CRM-positive patients. Survival analysis in relation to two gastrointestinal surgeons and a group of other surgeons showed survival improvements that paralleled a reduction in the rates of CRM involvement for the two gastrointestinal surgeons during the period of the study. No improvement in survival or reduction in rates of CRM involvement was seen in the group of other surgeons. CONCLUSIONS: These results show that CRM status may be used as an immediate predictor of survival after rectal cancer surgery and serves as a useful indicator of the quality of surgery. The frequency of CRM involvement can be used both for overall surgical audit and for monitoring the value of training programs in improving rectal surgery by individual surgeons. Its use in the current MRC CR07 study is valid and the best indicator of a requirement for further local therapy. PMID- 11923600 TI - Prognostic evaluation of stage B colon cancer patients is improved by an adequate lymphadenectomy: results of a secondary analysis of a large scale adjuvant trial. AB - OBJECTIVE: To determine if the extent of lymphadenectomy (number of recovered lymph nodes) was associated with long-term outcome in patients operated on for stage B and C colon cancer. SUMMARY BACKGROUND DATA: Lymphatic spreading is the main prognostic indicator in colon cancer patients, although the optimal extent of lymphadenectomy and its prognostic impact are still unknown. METHODS: In 3,648 patients (median follow-up 3.6 years) enrolled in two consecutive INTACC multicentric trials on adjuvant therapy for colon cancer, we studied the association of the number of recovered nodes with overall survival and relapse free survival by means of univariate and Cox regression analysis. RESULTS: The worst overall survival was related to ages > 65 (risk ratio [RR] = 1.30), higher grading (RR = 1.96). Better overall survival was related to female gender (RR = 0.80) and to higher number of recovered nodes (8-12 nodes, RR = 0.46, 13-17 nodes, RR = 0.76, nodes > or = 18, RR = 0.79). The same pattern was observed for relapse free survival. Longer overall and relapse free survival were related to a higher number of recovered nodes with P =.034 and P =.003 respectively (stratified analysis for absence or presence of positive nodes). Stage B patients with fewer than 7 nodes in the specimen had both shorter overall survival (P =.0000) and relapse free survival (P =.0016) than the other B patients. Outcome of stage C patients was not related to the number of recovered nodes (P =.28 and 0.12 respectively). The interaction test between stage of disease and number of recovered nodes was statistically significant (P =.017). CONCLUSIONS: Stage B patients with a small number of examined nodes may be understaged. Thus, these patients might be considered for adjuvant therapy because of their poorer life expectancy than other stage B patients. For stage C patients, the number of recovered nodes does not seem to affect long-term outcome. PMID- 11923601 TI - The effect of the surgeon and the pathologist on patient survival after resection of colon and rectal cancer. PMID- 11923603 TI - Transcontinental robot-assisted remote telesurgery: feasibility and potential applications. AB - OBJECTIVE: To show the feasibility of performing surgery across transoceanic distances by using dedicated asynchronous transfer mode (ATM) telecommunication technology. SUMMARY BACKGROUND DATA: Technical limitations and the issue of time delay for transmission of digitized information across existing telecommunication lines had been a source of concern about the feasibility of performing a complete surgical procedure from remote distances. METHODS: To verify the feasibility and safety in humans, the authors attempted remote robot-assisted laparoscopic cholecystectomy on a 68-year-old woman with a history of abdominal pain and cholelithiasis. Surgeons were in New York and the patient in Strasbourg. Connections between the sites were done with a high-speed terrestrial network (ATM service). RESULTS: The operation was carried out successfully in 54 minutes without difficulty or complications. Despite a round-trip distance of more than 14,000 km, the mean time lag for transmission during the procedure was 155 ms. The surgeons perceived the procedure as safe and the overall system as perfectly reliable. The postoperative course was uneventful and the patient returned to normal activities within 2 weeks after surgery. CONCLUSIONS: Remote robot assisted surgery appears feasible and safe. Teletransmission of active surgical manipulations has the potential to ensure availability of surgical expertise in remote locations for difficult or rare operations, and to improve surgical training worldwide. PMID- 11923604 TI - TP53 genotype but not p53 immunohistochemical result predicts response to preoperative short-term radiotherapy in rectal cancer. AB - OBJECTIVE: To evaluate and compare the predictive power of p53 gene analysis versus p53 immunohistochemical staining in terms of response to preoperative short-term radiotherapy using 25 Gy in operable rectal cancer. SUMMARY BACKGROUND DATA: Recent studies show that p53 may be a determinant of radiosensitivity being required for induction of apoptosis in case of radiation-induced DNA damage. METHODS: Preirradiation biopsy samples of 64 patients with rectal carcinoma were analyzed. Genetic alterations of the p53 gene were detected by complete direct sequencing of exons 2 to 10. Expression of the nuclear phosphoprotein p53 was assessed by immunohistochemical staining. Results were correlated with histopathology of resected specimens and follow-up data, respectively. RESULTS: Mutations of the p53 gene were present in 45% of tumors. Patients with a normal p53 gene had a significant survival advantage. Comparing pre- and postradiotherapy T category, a reduction was seen in patients with normal p53 genotype only. A mutant p53 genotype was highly specific in indicating stable disease concerning T category after irradiation. Protein overexpression was detected in 61%. Overexpression of the p53 protein was not related to survival or response. The concordance between immunohistochemistry and sequencing was only 0.51. CONCLUSIONS: The authors show that downstaging after short-term radiation may occur but is seen in tumors with normal p53 gene only. Moreover, p53 genotype but not p53 immunohistochemistry is predictive for response to preoperative short term radiotherapy and patient survival. PMID- 11923602 TI - Locoregional therapies for hepatocellular carcinoma: a critical review from the surgeon's perspective. AB - OBJECTIVE: This article reviews the current results of various locoregional therapies for hepatocellular carcinoma (HCC), with special reference to the implications for surgeons. SUMMARY BACKGROUND DATA: Resection or transplantation is the treatment of choice for HCC, but most patients are not suitable candidates. The past decade has witnessed the development of a variety of locoregional therapies for HCC. Surgeons are faced with the challenge of adopting these therapies in the management of patients with resectable or unresectable HCC. METHODS: A review of relevant English-language articles was undertaken based on a Medline search from January 1990 to August 2001. RESULTS: Retrospective studies suggested that transarterial chemoembolization is an effective treatment for inoperable HCC, but its perceived benefit for survival has not been substantiated in randomized trials, presumably because its antitumor effect is offset by its adverse effect on liver function. Nonetheless, it remains a widely used palliative treatment for HCC not amenable to resection or ablative therapies, and it also plays an important role as a treatment of postresection recurrence and as a pretransplant therapy for transplantable HCC. Better patient selection, selective segmental chemoembolization, and treatment repetition tailored to tumor response and patient tolerance may improve its benefit-risk ratio. Transarterial radiotherapy is a less available alternative that produces results similar to those of chemoembolization. Percutaneous ethanol injection has gained wide acceptance as a safe and effective treatment for HCCs 3 cm or smaller. Uncertainty in tumor necrosis limits its potential as a curative treatment, but its repeatability allows treatment of recurrence after ablation or resection of HCC that is crucial to prolongation of survival. Cryotherapy affords a better chance of cure because of predictable necrosis even for HCCs larger than 3 cm, but its use is limited by a high complication rate. There has been recent enthusiasm for heat ablation by microwave, radiofrequency, or laser, which provides predictable necrosis with a low complication rate. Preliminary data indicated that radiofrequency ablation is superior to ethanol injection in the radicality of tumor ablation. The advent of more versatile radiofrequency probes has allowed ablation of HCCs larger than 5 cm. Recent studies have suggested that combined transarterial embolization and heat ablation is a promising strategy for large HCCs. Thus far, no randomized trials comparing various thermoablative therapies have been reported. It is also uncertain whether a percutaneous route, laparoscopy, or open surgery affords the best approach for these therapies. Thermoablative therapies have been combined with resection or used to treat postresection recurrence, and they have also been used as a pretransplant therapy. However, the value of such strategies requires further evaluation. CONCLUSIONS: Advances in locoregional therapies have led to a major breakthrough in the management of unresectable HCC, but the exact role of the various modalities needs to be defined by randomized studies. Novel thermoablative techniques provide the surgeon with an exciting opportunity to participate actively in the management of unresectable HCC. Locoregional therapies are also useful adjuncts in the management of patients with resectable or transplantable disease. Hence, surgeons must be equipped with the latest knowledge and techniques of ablative therapy to provide the most appropriate treatment for the wide spectrum of patients with HCC. PMID- 11923606 TI - Increased adhesion molecule expression in serosal fibroblasts isolated from patients with inflammatory bowel disease is secondary to inflammation. AB - OBJECTIVE: To examine the expression of adhesion molecules by serosal and dermal fibroblasts in patients with inflammatory bowel disease. SUMMARY BACKGROUND DATA: The pathophysiologic process that leads to stricture formation in Crohn's disease (CD) is unknown. Serosal fibroblasts in these patients have an enhanced ability to contract collagen. This property may be reflected in fibroblast adhesion molecule expression, which in turn may be constitutive or secondary to the inflammatory process in patients with CD. METHODS: Fibroblasts were isolated from inflamed and macroscopically normal serosa of patients with CD or ulcerative colitis (UC) and from normal serosa of patients with colon cancer. Dermal fibroblasts were also isolated from the wound edge. Cell surface and whole cell expression of ICAM-1 were evaluated by flow cytometry and Western blot analysis, respectively. NFkappaB was measured by mobility shift assay in parallel experiments. Interleukin 1beta was added to the culture medium. RESULTS: Expression of ICAM-1 and NFkappaB, increased in patients with both CD and UC, was unaltered by interleukin 1beta. The whole cell concentration of ICAM-1 was greater in patients with CD than in patients with UC. Dermal fibroblasts did not display these features. CONCLUSIONS: Patients with inflammatory bowel disease display enhanced ICAM-1 expression in serosal fibroblasts but not dermal fibroblasts, indicating a secondary response to inflammation. PMID- 11923605 TI - Quantitative detection of disseminated free cancer cells in peritoneal washes with real-time reverse transcriptase-polymerase chain reaction: a sensitive predictor of outcome for patients with gastric carcinoma. AB - OBJECTIVE: To evaluate the feasibility of real-time reverse transcriptase polymerase chain reaction (RT-PCR) detection of free cancer cells in the peritoneal washes as a prognostic indicator for patients with gastric carcinoma. SUMMARY BACKGROUND DATA: Peritoneal lavage cytology (CY) is an excellent prognostic determinant but lacks sensitivity. This can be improved by using RT PCR to quantitate carcinoembryonic antigen (CEA) mRNA in peritoneal washes. METHODS: Peritoneal washes were obtained from 189 patients with gastric carcinoma during laparotomy. CEA mRNA levels and CEA/GAPDH mRNA ratios were quantified using a real-time PCR system with fluorescent hybridization probes. Receiver operating characteristic plots were used to determine which of these parameters should be used as a marker for the intraperitoneal cancer cells. The prognostic significance of its positivity was then evaluated by Kaplan-Meier curves, and its value as an independent prognostic factor was evaluated by multivariate analysis. RESULTS: The sensitivity and specificity of real-time RT-PCR with an optimal cutoff value were 80% and 94%; those for conventional cytology were 56% and 91%. The survival of 16 patients who were CY-PCR+ was poor and approached that of 35 CY+ patients. Recurrence as peritoneal carcinomatosis was frequent among PCR+ patients but rare for their PCR- counterparts. PCR+ was a significant independent prognostic factor, along with the presence of node metastasis and serosal invasion, but CY+ was not. CONCLUSIONS: Quantitative RT-PCR of peritoneal washes can replace cytologic examination as a tool for the sensitive evaluation of the risk of intraperitoneal recurrence in patients with gastric carcinoma. PMID- 11923607 TI - Association of color Doppler vascularity index and microvessel density with survival in patients with gastric cancer. AB - OBJECTIVE: The purpose of this study was to investigate the clinical usefulness of microvessel density (MVD) and an in vivo angiogenesis parameter, color Doppler vascularity index (CDVI), in patients with gastric cancer. SUMMARY BACKGROUND DATA: Many studies have reported a significant association between the degree of MVD-evaluated angiogenesis with the clinicopathologic factors and prognosis of patients with various solid tumors. All these studies were accomplished on tissue sections retrospectively obtained from surgical specimens. However, an in vivo method to assess tumor angiogenesis for human malignancies is highly desirable for diagnostic purpose, treatment planning, and follow-up. The CDVI is a new ultrasound parameter for evaluating in vivo angiogenesis, has a good correlation with status of lymph node metastasis in cervical carcinoma, and can predict distant metastasis and survival in colon cancer patients. Therefore, the CDVI may also be useful to assess in vivo angiogenesis in human gastric cancer. METHODS: A total of 79 patients with gastric cancer were enrolled in this study, and microvessel density was evaluated by using immunohistochemical staining of surgical specimens with anti-CD-34 antibody. Tumors were sonographically visible in 31 patients. The CDVI of each tumor was determined using transabdominal color Doppler ultrasound. The CDVI was defined as the ratio of the number of the colored pixels within a tumor section to the number of total pixels in that specific tumor section, and was calculated by using Encomate software (Electronic Business Machine Co. Ltd., Taipei, Taiwan). Correlation between MVD, CDVI and clinicopathologic factors and patient survival was studied. RESULTS: The MVD was significantly correlated with vascular invasion by multiple linear regression analysis. Although the survival of patients with high MVD (> 32) was significantly worse than those with low MVD (< 32) by univariate analysis, vascular invasion was an independent prognostic factor by Cox proportional hazard model. There was a linear correlation between CDVI and MVD (r =.495, P =.005). Moreover, in patients with a high CDVI (> 11%), the survival rate was significantly lower than that in those with low CDVI (< or = 11%, P =.005). None of the patients with high CDVI (> 11%) survived 2 years after curative resection. In addition to vascular invasion, the CDVI was another independent prognostic factor in the patients with stage III gastric cancer. CONCLUSIONS: Vascular invasion was an important prognostic indicator in gastric cancer. The high CDVI was a good preoperative indicator of early death in stage III gastric cancer patients. Thus, the CDVI may be helpful in selecting patients with gastric cancer for neoadjuvant chemotherapy and/or anti-angiogenic therapy. PMID- 11923608 TI - Human macrophage metalloelastase worsens the prognosis of pancreatic cancer. AB - OBJECTIVE: To evaluate the role of human macrophage metalloelastase (HME) in pancreatic cancer. SUMMARY BACKGROUND DATA: HME, a member of the human matrix metalloproteinase family, possesses elastolytic activity and is critical for the degradation of extracellular matrix proteins. Inasmuch as tumor invasion and metastasis formation require lysis of extracellular matrix, HME plays a critical role in both processes. METHODS: HME expression was analyzed by Northern blot analysis, reverse transcriptase-polymerase chain reaction, Western blot analysis, and immunohistochemistry in 39 pancreatic cancer tissues and 13 normal controls. The molecular data were related to clinicopathologic parameters and patient survival. RESULTS: In human pancreatic cancer, overexpression of HME mRNA was present in 25 of 39 pancreatic cancer tissues (64%) and in five pancreatic cancer cell lines. In contrast, low levels of HME mRNA expression were present in 13 normal pancreatic tissues samples. By Western blot analysis, high levels of HME were found in pancreatic cancer tissues and in the pancreatic cancer cell lines compared with the normal controls. Fifty-six percent of the cancer samples exhibited HME immunoreactivity in the cancer cells, and 63% in the stromal cells. Analysis of the survival data revealed that patients whose tumors exhibited HME mRNA overexpression lived significantly shorter compared with patients whose tumors did not overexpress HME. No relationship between HME expression and tumor stage, tumor grading, or presence of lymph node metastases was found. CONCLUSIONS: These findings indicate that HME participates in pancreatic cancer progression and that its presence worsens the prognosis. These data suggest a benefit of its inhibition in the treatment of pancreatic cancer. PMID- 11923609 TI - Accuracy of multiphasic helical computed tomography and intraoperative sonography in patients undergoing orthotopic liver transplantation for hepatoma: what is the truth? AB - OBJECTIVE: To determine the real value of liver imaging in cirrhosis by macro- and histomorphologic examination of the entire organ after orthotopic liver transplantation for hepatocellular carcinoma (HCC). SUMMARY BACKGROUND DATA: In comparative studies, a virtual sensitivity of up to 94% is described for helical computed tomography in HCC staging. The tumor detection rate of intraoperative ultrasonography (IOUS) is reported to be almost 100%. METHODS: This prospective observational study comprised 23 patients with HCC in cirrhosis admitted for orthotopic liver transplantation. Results of preoperative triphasic helical computed tomography (HCT) and IOUS were correlated with histopathologic results after 3-mm-slicing of the explanted liver. RESULTS: Overall, 179 liver segments were examined by HCT, IOUS, and MHM. Fifty-two malignant lesions and 10 dysplastic nodules were revealed by MHM. Using HCT, 13 HCCs could not be identified in 8 patients and 15 results were falsely positive in 10 patients. The detection rate of dysplastic nodes was 40% for HCT and 60% for IOUS. IOUS missed four HCCs in four patients and had six false-positive results in six patients. In a segment-based analysis, the overall accuracy of IOUS was significantly higher for IOUS (95.5%) versus HCT (89.9%). In the lesion-by-lesion analysis, the sensitivity was 92.3% for IOUS and 75.0% for HCT, with a significant difference. CONCLUSIONS: Correlation of explanted liver pathologic results offers precise evaluation of imaging modalities. The data of this histopathologically based study confirm that IOUS is significantly superior in staging HCC in cirrhosis versus CT, even after technical refinements through enhanced multiphasic high velocity helical scanning. PMID- 11923610 TI - Long-term results with multimodal adjuvant therapy and liver transplantation for the treatment of hepatocellular carcinomas larger than 5 centimeters. AB - OBJECTIVE: To determine the long-term results of liver transplantation for hepatocellular carcinoma (HCC) measuring 5 cm or larger treated in a multimodality adjuvant protocol. SUMMARY BACKGROUND DATA: Transplant has been established as a viable treatment of HCC measuring less than 5 cm, but the results for larger tumors have been disappointing. Several studies have shown promising preliminary results when combining transplant with preoperative transarterial chemoembolization and/or perioperative systemic chemotherapy in the treatment of advanced HCC that is not amenable to resection. However, follow-up in the studies has been limited and the number of patients has been small. METHODS: Beginning in October 1991, all patients with unresectable HCC measuring 5 cm or larger, as measured by computed tomography, were considered for enrollment in the authors' multimodality protocol. Entry criteria required that all patients be free of extrahepatic disease based on computed tomography scans of the chest and abdomen and bone scan and have a patent main portal vein and major hepatic veins on duplex ultrasonography. Patients received subselective arterial chemoembolization with mitomycin C, doxorubicin, and cisplatin at the time of diagnosis, repeated as necessary based on tumor response. Patients received a single systemic intraoperative dose of doxorubicin (10 mg/m(2)) before revascularization of the new liver and systemic doxorubicin (50 mg/m(2)) every 3 weeks as tolerated, for a total of six cycles, beginning on the sixth postoperative week. RESULTS: Eighty patients were enrolled; 37 were eventually excluded, due mainly to disease progression while on the waiting list, and 43 underwent liver transplant. Mean pathologic tumor diameter was 5.8 +/- 2.7 cm. Median follow-up of surviving transplanted patients was 55.1 +/- 24.9 months. There were two (4.7%) perioperative deaths. Median overall survival was significantly longer in transplanted patients (49.9 +/- 10.42 months) than in those who were excluded (6.83 +/- 1.34 months). Overall and recurrence-free survival rates in transplanted patients at 5 years were 44% and 48%, respectively. A tumor size larger than 7 cm and the presence of vascular invasion correlated significantly with recurrence. Recurrence-free survival at 5 years was significantly higher for the 32 patients with tumors measuring 5 to 7 cm (55%) than the 12 patients with tumors larger than 7 cm (34%). CONCLUSIONS: A significant proportion of patients with HCC measuring 5 cm or larger can achieve long-term survival after liver transplantation in the context of multimodal adjuvant therapy. Patients with tumors measuring 5 to 7 cm have significantly longer recurrence-free survival compared with those with larger tumors. PMID- 11923612 TI - Alteration of chemoattractant receptor expression regulates human neutrophil chemotaxis in vivo. AB - OBJECTIVE: To elucidate the mechanisms that regulate human neutrophil delivery in vivo, as well as the mechanisms that lead to observed reduction in polymorphonuclear (PMN) delivery to remote sites in septic patients. METHODS: Alterations in human PMN chemoattractant receptor expression and chemotactic function in vivo were evaluated in two distinct experiments: exudate PMNs (PMNs that have undergone transmigration to skin window blisters in controls) and septic PMNs (circulating PMNs from septic patients in the intensive care unit) were both separately compared with control circulating PMNs. RESULTS: Exudate PMNs displayed increased C5a receptors and C5a chemotaxis, and reduced interleukin-8 receptors (both IL-8 RA and IL-8 RB) and IL-8 chemotaxis. Septic PMNs displayed reduced C5a and IL-8 receptors and decreased C5a chemotaxis but no change in IL-8 chemotaxis. IL-8 but not C5a receptor gene expression decreased in parallel to receptor alteration. CONCLUSIONS: These results suggest that change in PMN chemoattractant receptor expression serves to regulate PMN chemotaxis in vivo; that exudate PMN chemotaxis depends more on C5a than IL-8; and that diminished chemoattractant receptors and chemotaxis in septic PMNs may explain decreased PMN delivery in these patients. PMID- 11923611 TI - Induction of tumor-specific cytotoxic T lymphocytes in cancer patients by autologous tumor RNA-transfected dendritic cells. AB - OBJECTIVE: To demonstrate the feasibility of inducing tumor antigen-specific immune responses in patients with metastatic cancer using total tumor RNA-loaded dendritic cells (DCs). SUMMARY BACKGROUND DATA: The authors have shown that DCs transfected with mRNA encoding defined tumor antigens induce tumor antigen specific T-cell responses in vitro and in vivo. There may be significant advantages to inducing immune responses against the entire repertoire of antigens expressed by a patient's autologous tumor. METHODS: RNA was extracted from a metastatic colon cancer and used to load autologous DCs. The DCs were coincubated with autologous T cells and the cytolytic activity of the T cells was assessed by the ability to lyse the autologous tumor cells. RNA was then extracted from a metastatic lung cancer and used to load autologous DCs, followed by four injections of the DC vaccine given every 4 weeks. Tumor antigen-specific cytotoxic T lymphocyte activity was then evaluated by testing peripheral blood mononuclear cells for their ability to lyse an antigen-expressing target. RESULTS: DCs transfected with the total RNA content of autologous tumor cells stimulated antigen-specific T-cell responses that are capable of recognizing and lysing autologous, primary tumor cells in vitro. Tumor-specific immune responses were induced in a patient with a carcinoembryonic antigen-expressing adenocarcinoma after immunization with autologous DCs transfected with total tumor RNA. CONCLUSIONS: DCs transfected with total tumor RNA may represent a method for inducing immune responses against the entire repertoire of tumor antigens of surgically resected malignancies. PMID- 11923613 TI - Impaired monocyte IL-12 production before surgery as a predictive factor for the lethal outcome of postoperative sepsis. AB - OBJECTIVE: To investigate whether monocyte paralysis resistant to interferon gamma (IFN-gamma) costimulation may exist before surgery and postoperative infection and may correlate with the outcome of postoperative sepsis. SUMMARY BACKGROUND DATA: Several studies have correlated monocyte paralysis during the course of sepsis with lethal outcome. Although the authors' previous work indicated that preoperative defects in monocyte interleukin (IL)-12 production are associated with the development of severe postoperative sepsis, the functional state of monocytes before surgery and infection and its significance for sepsis requires further analysis. METHODS: In a prospective study, monocyte functions of 1,113 consecutive patients were examined before major visceral surgery. Monocytes were isolated from peripheral blood and were stimulated in vitro with IFN-gamma and lipopolysaccharide. The secretion of IL-12 p70, IL-12 p40, IL-10, and tumor necrosis factor was measured. RESULTS: Preoperative monocyte secretion of IL-12 p70 and IL-12 p40 was significantly reduced in patients who developed lethal postoperative sepsis compared with sepsis survivors and patients with uneventful postoperative recovery. Moreover, preoperative monocyte IL-12 production was an independent predictive factor for the lethal outcome of postoperative sepsis by multivariate analysis. Preoperative monocyte IL-10 production was impaired in the sepsis group but did not correlate with death from sepsis. Preoperative monocyte tumor necrosis factor secretion was comparable between patients with uneventful recovery, sepsis survivors, and nonsurvivors. Thus, impaired preoperative monocyte IL-12 secretion in patients developing lethal postoperative sepsis did not result from an overproduction of IL-10 or from a generalized monocyte paralysis. The association between impaired preoperative monocyte IL-12 production and death from sepsis was also not explained by gender differences, underlying malignant disease, tumor type, neoadjuvant therapy, or age. CONCLUSIONS: These results identify a selective preoperative defect in monocyte IL-12 production as a predictive factor for the lethal outcome of postoperative sepsis. These data suggest that a partial preoperative monocyte paralysis severely impairs the host defense against postoperative infection, resulting in an increased risk of lethal sepsis. PMID- 11923615 TI - Variation in death rate after abdominal aortic aneurysmectomy in the United States: impact of hospital volume, gender, and age. AB - OBJECTIVE: To determine whether high-volume hospitals (HVHs) have lower in hospital death rates after abdominal aortic aneurysm (AAA) repair compared with low-volume hospitals (LVHs). SUMMARY BACKGROUND DATA: Select statewide studies have shown that HVHs have superior outcomes compared with LVHs for AAA repair, but they may not be representative of the true volume-outcome relationship for the entire United States. METHODS: Patients undergoing repair of intact or ruptured AAAs in the Nationwide Inpatient Sample (NIS) for 1996 and 1997 were included (n = 13,887) for study. The NIS represents a 20% stratified random sample representative of all U.S. hospitals. Unadjusted and case mix-adjusted analyses were performed. RESULTS: The overall death rate was 3.8% for intact AAA repair and 47% for ruptured AAA repair. For repair of intact AAAs, HVHs had a lower death rate than LVHs. The death rate after repair of ruptured AAA was also slightly lower at HVHs. In a multivariate analysis adjusting for case mix, having surgery at an LVH was associated with a 56% increased risk of in-hospital death. Other independent risk factors for in-hospital death included female gender, age older than 65 years, aneurysm rupture, urgent or emergent admission, and comorbid disease. CONCLUSIONS: This study from a representative national database documents that HVHs have a significantly lower death rate than LVHs for repair of both intact and ruptured AAA. These data support the regionalization of patients to HVHs for AAA repair. PMID- 11923614 TI - Inosine exerts a broad range of antiinflammatory effects in a murine model of acute lung injury. AB - OBJECTIVE: To investigate the effects of inosine on the acute lung inflammation induced by lipopolysaccharide (LPS) in vivo and on the activation and cytotoxicity elicited by proinflammatory cytokines on human lung epithelial (A549) cells in vitro. SUMMARY BACKGROUND DATA: Inosine is an endogenous purine recently shown to exert immunomodulatory and antiinflammatory effects. METHODS: Mice challenged with intratracheal LPS (50 microg) were treated after 1, 6, and 12 hours with inosine (200 mg/kg intraperitoneal) or vehicle. After 24 hours, bronchoalveolar lavage fluid was obtained to measure proinflammatory (tumor necrosis factor-alpha [TNF-alpha], interleukin [IL]-1beta, IL-6), and antiinflammatory (IL-10, IL-4) cytokines, chemokines (MIP-1alpha and MIP-2), myeloperoxidase activity and total cell counts, nitric oxide production, and proteins. Lung histology and immunohistochemical detection of 3-nitrotyrosine, a marker of nitrosative stress, were performed in inflated-fixed lungs. In vitro, cell viability and production of the chemokine IL-8 were evaluated in A549 cells stimulated with a mixture of cytokines in the presence or absence of inosine. RESULTS: Inosine downregulated the LPS-induced expression of TNF-alpha, IL-1beta, IL-6 and MIP-2 and tended to reduce MIP-1alpha, whereas it enhanced the production of IL-4. Total leukocyte counts, myeloperoxidase, nitric oxide production, and proteins were all significantly decreased by inosine. The purine also improved lung morphology and suppressed 3-nitrotyrosine staining in the lungs after LPS. Inosine attenuated the cytotoxicity and the expression of IL-8 induced by proinflammatory cytokines in A549 cells. CONCLUSIONS: Inosine largely suppressed LPS-induced lung inflammation in vivo and reduced the toxicity of cytokines in lung cells in vitro. These data support the proposal that inosine might represent a useful adjunct in the therapy of acute respiratory distress syndrome. PMID- 11923616 TI - Lipomas of the cord and round ligament. AB - OBJECTIVE: To determine the incidence, significance, and anatomy of spermatic cord and round ligament lipomas. METHODS: This was a retrospective review of 280 hernia repairs on 217 patients performed by a single surgeon (M.E.A.) from January 1996 to January 2000. The incidence of cord lipoma and relationship to inguinal hernia were evaluated. Further, when identified at the time of laparoscopic preperitoneal hernia repair, the anatomy of the lipomas was studied both at the time of surgery and again on review of videotapes. RESULTS: One hundred ninety-nine laparoscopic and 81 open inguinal hernia repairs were performed on 192 male patients and 25 female patients. Sixty-three lipomas of the cord were identified for an incidence of 22.5%. Overall, 18 cord lipomas were found in groins without hernias, and these were identified before surgery in 10 (2 by physical examination, 7 by groin ultrasound, and 1 by magnetic resonance imaging). The remaining nine were misidentified as a hernia before surgery. Fourteen of these patients presented with groin pain and four were asymptomatic. Forty-five lipomas were associated with hernias and were characterized as a hernia by examination in 43 instances. There were 32 (51%) cord lipomas associated with indirect hernias, 11 (17%) with direct hernias, and 1 each with pantaloon and femoral hernias. Nine lipomas were found in women, seven presenting with groin pain and six found without an associated peritoneal defect. Two patients presented with symptomatic cord lipomas after laparoscopic hernia repair. A lipoma of the cord is herniated fat that appears to originate from the retroperitoneal fat outside and posterior to the internal spermatic fascia and protrudes through the internal ring lateral to the cord. They are generally not visible by transperitoneal inspection unless manually reduced. CONCLUSIONS: Lipomas of the cord and round ligament occur with a significant incidence. They can cause hernia-type symptoms in the absence of a true hernia (associated with a peritoneal defect). They should be considered in the patient with groin pain and normal examination results. They can be easily overlooked at the time of laparoscopic hernia repair, and this can lead to an unsatisfactory result. PMID- 11923617 TI - Andre Toupet: surgeon technician par excellence. AB - Andre Toupet is best known for the posterior fundoplication that bears his name, currently used for the treatment of gastroesophageal reflux disease (GERD) or completing Heller's myotomy and subject today to intense discussions. This was not different in 1963, when Toupet proposed his technique at a time when the Nissen fundoplication was emerging as the treatment of choice for GERD. Behind the procedure, we discover a man with great surgical talent and meticulous attention to technical details who opposed criticism with hard work and strong family values. PMID- 11923620 TI - Resection of hepatocellular carcinoma with out preoperative tumor biopsy. PMID- 11923621 TI - Prospective comparison of stereotactic core biopsy and surgical excision as diagnostic procedures for breast cancer patients. PMID- 11923622 TI - Portal venous and enteric exocrine drainage versus systemic venous and bladder exocrine drainage of pancreas grafts. Clinical outcome of 40 consecutive transplants recipients. PMID- 11923623 TI - Intraabdominal infection several months following laparoscopic cholecystectomy. PMID- 11923624 TI - Developmental vasculopathies as manifestations of dysregulated histogenetic signal transduction pathways. PMID- 11923625 TI - Stroke and the child's brain: an overview of epidemiology, syndromes and risk factors. AB - Childhood stroke is emerging as a serious and frequent disorder. In contrast to adult stroke, the study of childhood stroke is in a very early stage of research development with no randomized controlled trials conducted to date outside of sickle cell stroke. The results of research in adults have limited applicability to children with stroke due to fundamental age-related differences in the neurological, cerebrovascular and coagulation systems. In recent years clinical and basic research studies have improved our understanding of childhood stroke. Population-based studies have resulted in an increased awareness of the frequencies and features of stroke syndromes in children. Cohort and case-control studies have led to an enhanced understanding of the multiple risk factors underlying childhood stroke. Advances in neuroimaging and in genetic and other laboratory testing approaches have resulted in an increased definition of stroke subtypes and risk factors. PMID- 11923626 TI - Arterial ischemic stroke in childhood: the role of plasma-phase risk factors. AB - The role of plasma-phase risk factors for stroke in the pediatric age group is presently unclear due to the lack of sufficiently large prospective studies, and due to the fact that these risk factors do not apply uniformly to newborns, children with sickle cell disease, and older children. Available evidence indicates that factor V Leiden, prothrombin 20210A, and lipoprotein (a) are all important in the pathogenesis of arterial ischemic stroke in older children, but the role of other plasma-phase risk factors remains uncertain. The contribution of these risk factors to newborn stroke and the stroke of children with sickle cell disease is similarly unclear, likely because the ischemia in affected children is predominantly due to nonhematologic perinatal events and erythrocyte adhesion to endothelium with obstruction of flow in the cerebral microcirculation, respectively. Evaluation of childhood stroke should, in our view, always be performed from the standpoint of the presenting clinical symptoms, diagnostic imaging, and determination of plasma-phase risk factors. Therapeutic anticoagulation and use of antiplatelet agents at present focus on the older child. PMID- 11923627 TI - Diseases of adenosine triphosphate synthesis in children. AB - There is an expanding understanding of primary genetic oxidative-phosphorylation disorders and the recognition of new multi-system clinical phenotypes in the energy metabolism diseases. Although initially recognized in association with mitochondrial DNA mutations, there is progress in the more laborious identification of nuclear DNA encoded genes relevant to mitochondrial structure and function. More pathogenic mitochondrial DNA and nuclear DNA mutations have been identified. Diagnosis of these disorders is often difficult and relies on a concurrence of findings, including recognition of a variety of clinical signs and symptoms, biochemical marker screening, electron transport chain enzyme measurements, and mitochondrial DNA or nuclear DNA mutation assay of genes relevant to mitochondrial structure, function or adenosine triphosphate metabolic pathways. Clinical diagnostic assessment now can be augmented by physiologic imaging techniques, including nuclear magnetic resonance spectroscopy and positron emission tomography. These capabilities should be increasingly helpful for studies of clinical progression and therapeutic intervention. Biologic studies, in families and patients, are beginning to address the factors of mitochondrial replication and segregation that underlie cellular/tissue heteroplasmy and clinical variability. Most epigenetic factors affecting organ specific and phenotypic variability, however, remain to be elaborated. PMID- 11923628 TI - Perinatal brain injury in the preterm and term newborn. AB - Major advances in understanding the cellular mechanisms of brain injury have presented a host of potential targets for intervention. This is particularly true of hypoxic-ischemic injury, the most important form of perinatal brain injury. As the window for effective clinical intervention may be particularly narrow in the fetus and newborn because of the often-delayed and subtle presentation of the onset of the insult, recent focus has been on defining and countering the more delayed mechanisms of brain injury. Recent insights into the mechanisms of oligodendrocyte injury and the role of inflammatory substances in perinatal brain injury are also discussed. PMID- 11923629 TI - Infectious and inflammatory disorders of the circulatory system and stroke in childhood. AB - The role of infectious and inflammatory causes of stroke is much more significant in children than in adults. Conversely, that of atherosclerotic disease, ischaemic heart disease and hypertensive haemorrhages has a lesser prominence in children. Bacterial meningitis caused by Streptococcus pneumoniae, Haemophilus influenzae, or Neiserria meningitidis has been known to cause stroke in children. The mechanism appears to be the spread of meningeal inflammation to involve the walls of intracranial vessels, resulting in arterial thrombosis with ischaemia or rupture with haemorrhage. Other infections caused by atypical bacterial agents such as Mycoplasma tuberculosis and viral agents such as varicella-zoster virus have also been well documented as causes of stroke. Non-infectious, inflammatory causes of stroke, such as collagen vascular disease and primary angiitis of the central nervous system, have been reported in children as well as adults. In this review, we will focus on recent advances in the field of childhood stroke caused by infectious and inflammatory disorders. PMID- 11923630 TI - Surgical approaches to vascular anomalies of the child's brain. AB - The population of pediatric patients with cerebrovascular disease presents a unique set of challenges to the neurosurgeon. The wide scope of pathology, including arteriovenous malformations, cavernous malformations and moyamoya disease, coupled with marked advances in diagnosis and treatment of these conditions over the past several years has resulted in a proliferation of the literature related to this subject. The present review provides an overview of current methods of surgical treatment for pediatric cerebrovascular diseases, focusing on indications and use of recent refinements of pre-existing surgical approaches, and newly evolved operative techniques developed to treat these entities. PMID- 11923631 TI - Seizure prediction and the preseizure period. AB - Beginning in the 1970s engineers designed systems to predict epileptic seizures based upon quantitative changes in the electroencephalogram, which they hypothesized began well in advance of clinical seizure onset. These efforts flourished in the 1990s, as independent laboratories demonstrated evidence of a 'preseizure period' up to 20 min prior to clinical symptoms in patients implanted with intracranial electrodes during evaluation for epilepsy surgery. Years later, clinical and laboratory experiments leave little doubt that a preseizure period exists in temporal lobe and perhaps other forms of epilepsy. Its existence, however, raises fundamental questions about what constitutes a seizure, what brain regions are involved in seizure generation, and whether discrete interictal, preictal, ictal and post-ictal physiologies exist, or blend together in a continuous process. Pressing milestones, necessary for clinical utility, are: (1) demonstrating prospective seizure prediction from prolonged human data sets, (2) elucidating mechanisms underlying seizure precursors and (3) implementing these algorithms on implantable hardware platforms. The notion of a preseizure state is catalyzing new clinical and basic science research, which has the potential to dramatically increase our understanding of epilepsy, and to generate exciting new therapies for patients. PMID- 11923632 TI - The role of mitochondria in epileptogenesis. AB - Mitochondrial dysfunction has gained considerable interest as a potential cause of epileptic seizures and therapy-resistant forms of severe epilepsy. Impairment of mitochondrial function has recently been observed in the seizure focus of human and experimental epilepsy. Additionally, a broad variety of mutation of mitochondrial DNA leading to the inhibition of mitochondrial respiratory chain or directly of mitochondrial adenosine triphosphate synthesis in epileptogenic areas of the human brain has been associated with epileptic phenotypes. Since mitochondrial oxidative phosphorylation provides the major source of adenosine triphosphate in neurons, and mitochondria participate in cellular Ca2+ homeostasis they can modulate neuronal excitability and synaptic transmission. Furthermore, mitochondria are intimately involved in pathways leading to the neuronal cell death characteristic for the areas of epileptogenesis. PMID- 11923633 TI - Epilepsy surgery within the temporal lobe and its short-term and long-term effects on memory. AB - In patients with pharmacologically intractable, complex-partial seizures, surgical excision of the involved temporal lobe may have significant therapeutic benefit. Resection of mesial structures of the temporal lobes, however, entails a significant risk of decreased memory function. Recent advances in the assessment of memory changes following temporal lobectomy surgery emphasize the complexity of subjective ratings of memory functioning in this population. Neuroimaging tools useful in the diagnostic evaluation of epilepsy surgical candidates have now been shown to be useful in predicting memory change in the post-surgical period. Functional magnetic resonance imaging appears to provide significant information regarding hemispheric representation of language in the temporal lobe epilepsy patient, and the use of this technique to predict memory status following surgery appears promising. Clinical studies involving patients who had temporal lobectomy surgeries have also revealed changes in emotional learning related to the degree of amygdala involvement. Moreover, there is increasing evidence to suggest that differential changes in emotional learning occur among patients with right versus left temporal lobe resections. PMID- 11923634 TI - When is positron emission tomography really necessary in epilepsy diagnosis? AB - Positron emission tomography can be used for localization of epileptic foci, and preoperative functional mapping. Rapid improvements in magnetic resonance imaging, however, have restricted the need for positron emission tomography to a minority of patients who have unrevealing magnetic resonance imaging scans. Positron emission tomography will continue to be of value in investigations of the pathophysiology of seizure disorders. PMID- 11923636 TI - Vagus nerve stimulation: where are we? AB - Now nearly 5 years post-approval, vagus nerve stimulation has emerged as a major non-pharmacological treatment for epilepsy. The place of vagus nerve stimulation among antiepileptic drugs and other surgical therapies is still evolving. This review evaluates the role of vagus nerve stimulation in light of recently published research of its mechanism(s) of action, long-term efficacy, safety and tolerability, and application to other disorders besides epilepsy. PMID- 11923635 TI - Rasmussen's encephalitis: a role for autoimmune cytotoxic T lymphocytes. AB - The present review describes advances in Rasmussen's encephalitis (also known as Rasmussen's syndrome), an unihemispheric intractable epileptic disease with persistent inflammation. Specific attention is given to the recent recognition of cytotoxicity by CD8+/granzyme-B-positive T lymphocytes as a new pathogenic mechanism of neuronal damage. PMID- 11923637 TI - Treating patients with psychogenic non-epileptic seizures. AB - While the diagnostic features of psychogenic non-epileptic seizures have been better characterized in recent years, comparatively little is written about management. This review provides guidance to clinicians involved in the treatment of patients with psychogenic non-epileptic seizures and generates ideas for future research. It summarizes the recent literature specifically dealing with the treatment of such seizures and draws on the wider psychiatric literature on effective treatments for patients with other medically unexplained symptoms. PMID- 11923638 TI - Safety and pharmacokinetics of p792, a new blood-pool agent: results of clinical testing in nonpatient volunteers. AB - RATIONALE AND OBJECTIVES: To evaluate the safety and pharmacokinetics of P792, a new macromolecular blood-pool agent for magnetic resonance imaging (MRI), in nonpatient volunteers. METHODS: This was a single blind, placebo-controlled, ascending-dose study in 32 healthy male volunteers, randomized to receive a single intravenous dose of P792 (0.0065, 0.013, 0.026, and 0.039 mmol/kg). The safety controls consisted of complete pre- and postdose physical examinations, measurement of vital signs, clinical laboratory investigations, and monitoring of adverse events (up to 22 days after injection). For pharmacokinetic analysis, the determination of P792 was performed using the ICP-MS technique for blood and urine samples up to 22 days. RESULTS: No serious adverse events occurred during the study. There were no clinically significant changes in vital signs, or clinical laboratory findings. P792 blood half-life, distribution volume, and renal clearance are consistent with the definition of a rapid clearance blood pool agent (RCBPA) as defined previously. CONCLUSION: P792 appeared to be a safe and well-tolerated RCBPA in nonpatient subjects. Phase II studies will be conducted to evaluate the efficacy of the blood-pool agent for vascular, perfusion, and permeability imaging in MRI. PMID- 11923639 TI - Monomer-coated very small superparamagnetic iron oxide particles as contrast medium for magnetic resonance imaging: preclinical in vivo characterization. AB - RATIONALE AND OBJECTIVES: Preclinical in-vivo characterization of a newly developed MR contrast medium consisting of very small superparamagnetic iron oxide particles (VSOP) coated with citrate (VSOP-C184). METHODS: VSOP-C184 (core diameter: 4 nm; total diameter: 8.6 nm; relaxivities in water at 0.94 T (T1) 20.1 and (T2) 37.1 l/[mmol*sec]) was investigated to determine its pharmacokinetics, efficacy, acute single dose toxicity, repeated dose toxicity, and genotoxicity. RESULTS: The plasma elimination half-life at 0.045 mmol Fe/kg was 21.3 +/- 5.5 minutes in rats and 36.1 +/- 4.2 minutes in pigs, resulting in a T1-relaxation time of plasma of < 100 milliseconds for 30 minutes in pigs. The particles are mainly cleared via the phagocytosing system of the liver. MR angiography at a dose of 0.045 mmol Fe/kg shows an excellent depiction of the thoracic and abdominal vasculature in rats and of the coronary arteries in pigs. The LD50 in mice is > 17.9 mmol Fe/kg. A good tolerance and safety profile was found. CONCLUSIONS: The experiments indicate, that VSOP-C184 may be a well tolerated and safe contrast medium for MR imaging that can be effectively used for MR angiography including visualization of the coronary arteries. PMID- 11923641 TI - Magnetic resonance imaging of nude mice with heterotransplanted high-grade squamous cell carcinomas: use of a low-loaded, covalently bound Gd-Hsa conjugate as contrast agent with high tumor affinity. AB - RATIONALE: Malignant tumors often show an increased uptake and metabolism of plasma proteins, especially albumin. OBJECTIVES: Determine whether the accumulation of low loaded Gd-albumin improves visualization of malignant tumors by MRI. METHODS: Twelve nude mice with heterotransplanted squamous cell carcinomas were studied. The signal intensity of tumor, blood, liver, kidney and muscle tissue was studied in MR images after application of Gd-albumin during a period of 144 hours. MRI results were histologically correlated after simultaneously injection of Gd- and fluorescein-labeled albumins in 9 nude mice. RESULTS: Although liver and kidney had a maximum increase in signal intensity within 30 minutes, tumors showed a delayed 51% increase in the 24 hours after application. Histologic and fluorescence evaluation demonstrated albumin localization in tumors predominantly in stroma and necroses. CONCLUSIONS: Gd albumin is efficiently accumulated in SCC transplants. MRI with low loaded Gd albumin may offer relevant opportunities for recognizing tumors sensitive to a therapy with cyostic drug-labeled albumins. PMID- 11923640 TI - Identifying tumor vascular permeability heterogeneity with magnetic resonance imaging contrast agents. AB - RATIONALE AND OBJECTIVES: Dynamic contrast enhanced (DCE) MR mammography (MRM) uses tumor capillary density differences for prognosis. The heterogeneous response of permeability-surface area products (PS = Kp<-->t) was examined in mammary tumors, as a function of contrast agent size, to determine what effect ROI size might have on PS and prognosis. METHODS: DCE FLASH signal intensities were converted to gadolinium concentrations by a standard curve, which was fitted by a two-compartment model for the tumor's extravascular extracellular space (EES) volume fraction (ve), and the tumor volume normalized transfer rate between plasma and EES (Kp<-->t/VT). RESULTS: For Gd-DTPA ve = 9% to 13% Kp<-->t/VT = 0.01 to 0.06 minutes-1, and the macromolecular agent, PAMAM-TU-DTPA G = 4 ve = 0.8% to 1% Kp<-->t/VT = 0.008 to 0.04 minutes(-1). Significant differences in Kp< ->t/VT for local regions were found for both agents relative to the whole tumor and the macromolecular agent had greater dynamic range. CONCLUSIONS: Smaller ROI values or pixels should yield more accurate assessment of neovascularization. PMID- 11923642 TI - Biodistribution of a 153 Gd-folate dendrimer, generation = 4, in mice with folate receptor positive and negative ovarian tumor xenografts. AB - RATIONALE AND OBJECTIVES: An important characteristic of targeted contrast agents is how they are tolerated in a biologic environment and their localization in the surrounding tissues in addition to target tissue. We evaluate the biodistribution of a gadolinium Gd 153-folate-dendrimer in high affinity folate-receptor (hFR) positive and negative ovarian tumor xenografts. METHODS: The 153Gd-folate dendrimer chelate was prepared by exchanging 153Gd with nonradioactive gadolinium for 1 week, followed by extensive filtration. Athymic mice with hFR-positive (n = 3) and negative tumors (n = 3) were injected intravenously and counted using a whole-body counting system with a 80 to 150 keV counting window. RESULTS: The hFR positive tumors accumulate 3.6% +/- 2.8% injected dose/g, whereas only background counts were found in hFR-negative tumors. The folate-dendrimer's tumor-to-blood ratio of 12.6, in hFR-positive tumors, was approximately 5.7 to 17.0 fold better than those obtained with monoclonal antibodies targeted to the folate receptor. CONCLUSIONS: Biodistribution studies confirm previous MRI findings and show that the accumulation of the folate-dendrimer requires the expression of the hFR. PMID- 11923643 TI - Seeking consensus: contrast ultrasound in radiology. PMID- 11923644 TI - A physiologic flow chamber model to define intravascular ultrasound enhancement of fibrin using echogenic liposomes. AB - RATIONALE AND OBJECTIVES: Echogenic immunoliposomes (ELIP) for enhancement of vasoactive and pathologic components of endothelium and atherosclerosis have been developed. A physiologic flow chamber model has been developed to define intravascular ultrasound enhancement of a fibrin interface. METHODS: A IgG ELIP was used, which nonspecifically associated with fibrin, to demonstrate the suitability of this model. With varying doses of IgG ELIP, the fibrin wells were imaged at 1, 2, 4, 6, and 9 minutes. RESULTS: IgG ELIP enhanced fibrin versus saline (P < 0.005) was visible at 1 minute, lasted at least 9 minutes, and at 6 minutes the interface enhanced 27% +/- 6.1%. Enhancement was caused by increases in interface thickness and brightness. Enhancement increased with dose up to 8 mg lipid (n = 4 per time point). CONCLUSION: This model can quantitate the components of IVUS enhancement of an interface produced by ELIP. This model may allow for further development and understanding of ELIP and other targeted ultrasound contrast agents. PMID- 11923645 TI - Effect of iomeprol on rat hippocampal slice synaptic transmission: comparison with other X-ray contrast agents. AB - RATIONALE AND OBJECTIVES: All contrast agents should be neurologically safe because although some are not indicated for procedures, such as myelography, just the same they may come in contact with nervous tissue during contrast-enhanced imaging. This is because even when they are intravascularly injected, the presence of undiagnosed blood-brain barrier damage may allow them to penetrate the brain barrier. In the present study, we investigated the neurologic safety of iomeprol by studying in vitro its potential effects on the central nervous system (CNS) synaptic transmission. Other widely used x-ray contrast agents were also assessed for comparative purposes. METHODS: CNS synaptic transmission was evaluated in terms of evoked field potentials recorded from the pyramidal region of rat hippocampal slices. The field potentials were evoked by electrical stimulation of the Schaffer collateral pathway. The effects of the contrast agents were evaluated in terms of number and amplitude of population spikes (PS) and as the maximal slope of the excitatory postsynaptic potentials (EPSP). The contrast agents were tested at final concentrations of 3, 10, and 30 mg(iodine)/mL in iso-osmolal condition with respect to artificial cerebrospinal fluid (CSF). RESULTS: Iomeprol, like ioversol, principally exerted a mild inhibitory effect on CNS synaptic transmission, an effect that was preceded by a weak, transient excitation. Iopentol exerted a rapid and complete inhibition of synaptic transmission without showing any excitatory effects. Iobitridol, though belonging to the nonionic monomeric class, exerted, surprisingly, an epileptogenic action at the highest concentration, whereas its inhibitory action was slow and mild. Diatrizoate, as expected, exerted an epileptogenic activity even at the lowest concentration, followed by a marked inhibitory action. Ioxaglate, as expected because it is an ionic though dimeric contrast agent, exerted an epileptogenic action at the intermediate concentration, whereas it barely demonstrated an inhibitory effect at all. All the contrast agent effects observed in the study reversed or tended to reverse during washout. CONCLUSIONS: Even taking in account the limitation because of the use of an in vitro approach and high contrast agent concentrations, we can conclude that the positive neuro tolerability of iomeprol is further confirmed by this model as it proved to be devoid of epileptogenic activity and, among the contrast agents exhibiting inhibitory action, it was the contrast agent with the least amount of activity. In addition, contrary to that generally reported in the literature, nonionic, low osmolal contrast agents are not all identical in their neuro-tolerability when assessed in the rat hippocampal slice model. PMID- 11923646 TI - A surface-modified chylomicron remnant-like emulsion for percutaneous computed tomography lymphography: synthesis and preliminary imaging findings. AB - RATIONALE AND OBJECTIVES: To assess a surface-modified emulsion as a percutaneous CT lymphographic agent in normal dogs. METHODS: An iodinated chylomicron remnant like microemulsion was formulated with a mean particle size of 91.3 nm and an iodine concentration of 91 mg I/mL. Contrast material (2 mL) was injected into the subcutaneous tissues of the metatarsus and metacarpus of six normal dogs to enhance popliteal and cervical lymph nodes, respectively. CT images were acquired at 0, 15, 30, 45, 60, 240, 480, and 1440 minutes. RESULTS: Significant lymph node enhancement occurred in as little as 15 minutes after injection and persisted at least 8 hours. Node opacification was most pronounced at 1 to 4 hours postinjection and exceeded 200 HU in some nodes (precontrast attenuation = 45 HU). Marked enhancement of popliteal efferent lymphatics and of iliac and sacral node groups also occurred indicating distribution to second order nodes. Attenuation of enhanced nodes reverted to precontrast levels by 24 hours. CONCLUSION: The new surface-modified, chylomicron remnant-like emulsion provided marked, selective enhancement of targeted lymph nodes after subcutaneous administration. Moreover, the formulation produced significant opacification of more distant node groups from a single injection. PMID- 11923647 TI - Prospective observations of emerging psychosis. AB - Because delays in treatment of psychosis may be associated with poorer outcomes, intervention focus has shifted to the prodromal phase of illness. However, knowledge about this phase has been limited to retrospective reconstructions of symptoms once psychosis is already present. The following article offers a new, prospective view of the development of schizophrenia starting from the late prodromal phase of illness. As we use the term, late prodromal phase of illness means at imminent risk of conversion to schizophrenia. In this article, entry, conversion, and discharge data are presented on a sub-population of eight patients who received treatment for psychosis at onset while participating in a prospective double-blind placebo-controlled treatment study of the prodrome to psychosis. Through disguised case examples, this article then focuses on the course of illness progression in three of eight cases who converted to schizophreniform psychosis while randomized on either active medication or placebo. Discussion focuses on the dimensional quality of the transition both from the prodrome to psychosis and from psychosis to recovery. PMID- 11923648 TI - Insight and work performance in schizophrenia. AB - Research has linked impaired insight in schizophrenia to poorer medication compliance and poorer treatment outcome. The current study attempts to replicate previous findings that impaired insight is related to deficits in work function. To examine this question, 121 participants with schizophrenia or schizoaffective disorder enrolled in vocational rehabilitation were classified as having unimpaired (N = 65), or impaired (N = 56) insight. Next, participants were assigned a work placement and their work performance assessed on the third, fifth, and seventh weeks of work by using the Work Behavior Inventory. Among the 85 participants who completed these weeks of work, a multivariate analysis of variance and subsequent analysis of variance showed participants with impaired insight had significantly poorer ratings of work quality, work habits, cooperativeness, and personal presentation. When measures of global intelligence and executive function were entered as covariates in individual analysis of covariance, groups differed on measures of cooperativeness and personal presentation. Implications for rehabilitation are discussed. PMID- 11923649 TI - Young adults with attention deficit hyperactivity disorder: subtype differences in comorbidity, educational, and clinical history. AB - The present study sought to examine subtype differences in comorbidity and in antisocial, educational, and treatment histories among young adults (ages 17-27) with attention deficit hyperactivity disorder (ADHD). Comparisons were made between ADHD Combined Type (ADHD-C; N = 60) and Predominantly Inattentive Type (ADHD-I; N = 36) relative to each other and to a community control group of 64 adults. Both ADHD groups had significantly less education, were less likely to have graduated from college, and were more likely to have received special educational placement in high school. Both groups also presented with a greater likelihood of dysthymia, alcohol dependence/abuse, cannabis dependence/abuse, and learning disorders, as well as greater psychological distress on all scales of the SCL-90-R than the control group. Both ADHD groups were more likely to have received psychiatric medication and other mental health services than control adults. In comparison with ADHD-I, adults with ADHD-C differed in only a few respects. The C-type adults were more likely to have oppositional defiant disorder, to experience interpersonal hostility and paranoia, to have attempted suicide, and to have been arrested than the ADHD-I adults. These results are generally consistent with previous studies of ADHD in children, extend these findings to adults with ADHD, and suggest that the greater impulsivity associated with the ADHD-C subtype may predispose toward greater antisocial behavior and its consequences than does ADHD-I type in adults. PMID- 11923650 TI - Science-based policy for psychosocial interventions in refugee camps: a Cambodian example. AB - No longer are the high rates of psychiatric morbidity associated with mass violence in refugee populations invisible to the humanitarian assistance community. However, identification of mental health risk and protective factors that can be utilized by policy planners is still lacking. The objective of this report is to provide an analytic approach to determining these factors. A description is provided from the first large-scale epidemiological study of Cambodian refugees confined to the Thailand-Cambodian border in the 1980s and 1990s. The original data from this study are reanalyzed to evaluate the mental health impact of psychosocial factors subject to the influence of camp authorities, such as opportunities in the refugee camp environment and personal behaviors, in addition to trauma. The results suggest the extraordinary capacity of refugees to protect themselves against mental illness despite horrific life experiences. The recommendation emerges for refugee policy makers to create programs that support work, indigenous religious practices, and culture-based altruistic behavior among refugees. As refugee mental health policy receives increasing attention from the international community, it must consist of recommendations and practices based on scientific analysis and empirical evidence. PMID- 11923651 TI - Peritraumatic dissociation and physiological response to trauma-relevant stimuli in Vietnam combat veterans with posttraumatic stress disorder. AB - A recent study found that female rape victims with acute posttraumatic stress disorder (PTSD) who received a high score on the Peritraumatic Dissociative Experiences Questionnaire exhibited suppression of physiological responses during exposure to trauma-related stimuli. The goal of our present study was to test whether the same relationship holds true for male Vietnam combat veterans with chronic PTSD, using secondary analyses applied to data derived from a Veteran's Affairs Cooperative Study. Vietnam combat veterans (N = 1238) completed measures to establish combat-related PTSD diagnostic status, extent of PTSD-related symptomatic distress, and presence of dissociative symptoms during their most stressful combat-related experiences. Extreme subgroups of veterans with current PTSD were classified as either low dissociators (N = 118) or high dissociators (N = 256) based on an abbreviated version of the Peritraumatic Dissociative Experiences Questionnaire. Dependent variables reflected subjective distress along with heart rate, skin conductance, electromyographic, and blood pressure data when responding to neutral and trauma-related audiovisual and imagery presentations. Veterans in the current PTSD group had significantly higher dissociation scores than did veterans in the lifetime and never PTSD groups. Among veterans with current PTSD, high dissociators reported greater PTSD-related symptomatic distress than did low dissociators, but the groups did not differ with respect to physiological reactivity to the trauma-related laboratory presentations. Our results replicate the previously reported relationship between peritraumatic dissociation and PTSD status in Vietnam combat veterans. However, we found no association between peritraumatic dissociation and the extent of physiological responding to trauma-relevant cues in male veterans with chronic combat-related PTSD. PMID- 11923652 TI - Reliability and validity of the Japanese-language version of the impact of event scale-revised (IES-R-J): four studies of different traumatic events. AB - The authors developed the Japanese-language version of the Impact of Event Scale Revised (IES-R-J) and investigated its reliability and validity in four different groups: workers with lifetime mixed traumatic events, survivors of an arsenic poisoning case, survivors of the Hanshin-Awaji earthquake, and survivors of the Tokyo Metro sarin attack. Evidence includes retest reliability and internal consistency of the IES-R-J. Posttraumatic stress disorder (PTSD) and partial PTSD cases indicated significantly higher scores than non-PTSD cases. The IES-R-J can be a useful self-rating diagnostic instrument particularly for survivors with PTSD symptoms as a clinical concern (PTSD + partial PTSD) by using a 24/25 cutoff in total score. In analysis of scale structure, the majority of intrusion and hyperarousal items were subsumed under the same cluster, whereas avoidance items made up a separate cluster. Female patients indicated higher scores than male patients. A negative weak correlation between age and the score was found only among female earthquake survivors. The IES-R-J can be used as a validated instrument in future international comparative research. PMID- 11923653 TI - Inpatient versus day hospital treatment for chronic, combat-related posttraumatic stress disorder: a naturalistic comparison. AB - This study adopted a quasi-experimental design to compare the treatment outcomes of inpatient-outpatient programs and day hospital programs for chronic, combat related posttraumatic stress disorder. Data were drawn from 202 Vietnam veterans who had completed treatment at four programs across Australia. The veterans were assessed on a range of psychological and social variables at intake to the programs and followed up at 3 and 9 months after discharge. A significant main effect was found for time, with veterans from both program models demonstrating improvements that were maintained over the 9-month follow-up period. Group by time effects were not significant, which suggests that inpatient-outpatient programs are not more efficacious than the less expensive day hospital alternatives. In line with current mental health policy directions, the current study lends broad support to the recommendation that treatment services for veterans with posttraumatic stress disorder be delivered in the least restrictive environment. PMID- 11923654 TI - Do panic symptoms during periods of remission predict relapse of panic disorder? AB - This study investigated the relationship between panic symptoms during remission and subsequent relapse of panic disorder. Research subjects were 169 individuals, enrolled in the Harvard/Brown Anxiety Research Project, who were in episodes of panic disorder at intake and remitted during the 8-year follow-up period. Panic symptoms during remission were examined as predictors of panic disorder relapse. For the relapses that did occur, we report the level of symptomatology during the previous 4 weeks. Depending on symptom severity and whether or not the relapse included agoraphobia, panic symptoms during remission were associated with an approximate two- to threefold increased risk of relapse. However, on examination of symptoms during the 4 weeks before relapse, we found that the majority of relapses were preceded by no panic symptoms. Thus, although panic symptoms during remission may indicate an increased risk of subsequent relapse, the absence of symptoms during remission does not indicate that relapse is unlikely. PMID- 11923655 TI - Assessing aggression severity with the revised staff observation aggression scale. PMID- 11923656 TI - Executive and motor skill functioning among cocaine-dependent schizophrenics and non-drug-abusing schizophrenics. PMID- 11923658 TI - Preoperative magnetic resonance imaging screening for a surgical decision regarding the approach for anterior spine fusion at the cervicothoracic junction. AB - STUDY DESIGN: In a study investigating the correlation between a set of designed criteria and judgments of surgical experience, 100 cervical magnetic resonance images from different patients were used. OBJECTIVES: To demonstrate reliable and reproducible anatomic measurements that can aid spine surgeons in selecting surgical approaches for anterior spine fusion in the cervicothoracic region. SUMMARY OF BACKGROUND DATA: Surgical approaches to the cervicothoracic junction vary among surgeons. Whereas sternotomy provides maximum exposure, less extensive approaches are preferred to minimize surgical trauma, provided surgical goals are not compromised. No quantitative criteria currently exist to determine before surgery the least invasive surgical approach for sufficiently exposing pertinent anatomy. METHODS: Thirteen geometric variables designed to be clinically practical and to expose important anatomic relations were used to evaluate 100 sagittal scout cervical magnetic resonance image sequences. An experienced spine surgeon independently rated each image for the most appropriate surgical approach to the C5-T2 region. The ratings were tested for interrater reliability using a second spine surgeon. After testing for interrater and intrarater reliability, the geometric measurements were correlated with the surgeon's selected surgical approaches for each intervertebral segment (P < 0.05). RESULTS: Instrument manubrial thoracic distances, reflecting standardized heights of intervertebral discs above or below the superior tip of the manubrium, were the most reliable, reproducible, and correlative with the choice of surgical approach. All the measurements but one, the instrument manubrial thoracic distance for T1/T2, demonstrated interrater and intrarater reliability, with an interclass correlation of at least 0.70. The primary surgeon-investigator indicated the anterior approach with sternotomy (n = 3) or the transverse cervical approach (n = 97) for the C7/T1 exposure, and the anterior approach with sternotomy (n = 43) or the transverse cervical approach (n = 57) for the T1/T2 exposure. The interrater questionnaire reliability results indicated statistical agreement between the primary surgeon-investigator and the second cervical spine surgeon at all vertebral segments evaluated. Instrument manubrial thoracic distances showed the strongest significant correlation with the surgical approach, demonstrating a statistical power of 1. For the C7/T1 exposure, the instrument manubrial thoracic distance for C7/T1 was 1.9 +/- 2 cm (95% confidence interval [CI] = 1.41 to 2.22) for the transverse cervical approach, and -3.3 +/- 1.3 cm (95% CI = -4.79 to 1.75)] for the anterior approach with sternotomy. The instrument manubrial thoracic distance measurements for C5/C6, C6/C7, and T1/T2 also showed nonoverlapping 95% confidence intervals for the transverse cervical versus the anterior approach with sternotomy for the C7/T1 exposure. For the T1/T2 exposure, all four instrument manubrial thoracic distance measurements again showed statistically significant differences between approaches, with nonoverlapping 95% confidence intervals and a statistical power of 1. In addition, the measurements elaborating the anterior-to-posterior distance of the thoracic outlet and the measurements of the angle between the planes of the intervertebral disc and the sternum also showed statistically significant differences between approaches for the T1/T2 segment, with a statistical power of at least 0.9. CONCLUSIONS: Strong correlations exist between objective measurements and the choice of surgical approach for anterior spine fusion. Among investigated anatomic relations, the instrument manubrial thoracic distance correlated most reliably with the surgeons' choice of the anterior approach. Such objective measurements represent tools that cervical spine surgeons can use to determine the surgical approach. PMID- 11923659 TI - The effect of cage stiffness on the rate of lumbar interbody fusion: an in vivo model using poly(l-lactic Acid) and titanium cages. AB - STUDY DESIGN: A goat interbody fusion model using poly-(L-lactic acid) and titanium cages was designed to evaluate the effect of cage stiffness on lumbar interbody fusion. OBJECTIVE: To investigate the effect of cage stiffness on the rate of interbody fusion. SUMMARY OF BACKGROUND DATA: Various types of cages considerably exceed the stiffness of vertebral bone, which ultimately may lead to postoperative complications. To avoid these complications, poly-(L-lactic acid) cages with limited stiffness have been designed. The mechanical integrity of the cages remains intact for at least 6 months. METHODS: Interbody fusions were performed at L3-L4 of 15 Dutch milk goats, and one of three cages was randomly implanted: 1) a titanium cage (n = 3), 2) a stiff poly-(L-lactic acid) cage (n = 6), or 3) a flexible poly-(L-lactic acid) cage (n = 6). Interbody fusion was assessed radiographically by three independent observers 3 and 6 months after surgery. RESULTS: At 3 months, all the poly-(L-lactic acid) specimens showed ingrowth of new bone, but with radiolucency in the fusion mass. At 6 months, solid arthrodesis was observed in four of six poly-(L-lactic acid) specimens, advanced ingrowth in one specimen, and infection in one specimen. Titanium cages showed ingrowth of bone, but with radiolucency in the fusion mass. Interbody fusion using poly-(L-lactic acid) cages showed a significantly higher rate statistically (P = 0.016) and more complete fusion than titanium cages of the same design. CONCLUSIONS: The reduced stiffness of poly-(L-lactic acid) cages showed enhanced interbody fusion, as compared with titanium cages after 6 months. Bioabsorbable poly-(L-lactic acid) cages thus may be a viable alternative to current interbody cage devices, thereby avoiding the concomitant problems related to their excessive stiffness. However, the bioabsorbability of the poly-(L-lactic acid) cages awaits investigation in a long-term study currently underway. PMID- 11923660 TI - Asymmetrical flexible tethering of spine growth in an immature bovine model. AB - STUDY DESIGN: A 12-week bovine survival study of tethering anterior spine growth that included untethered control subjects. OBJECTIVE: To determine the effects that a flexible cable attached to the anterolateral aspect of the thoracic spine has on spine growth in rapidly growing calves. SUMMARY OF BACKGROUND DATA: The search for a way to correct scoliosis without the use of an arthrodesis continues in an attempt to maintain normal spine mobility. Experience in the hemiepiphyseal stapling of long bones has provided a background rationale for attempting growth modulation in the spine. It is postulated that a mechanical tether to the anterior and lateral growth of the spine in a growing child with scoliosis may allow spontaneous correction of sagittal and coronal plane deformity obviating the need for an arthrodesis. METHODS: Eight calves (age, 3-4 weeks; weight, 47 +/ 4.6 kg) underwent right-side thoracotomies exposing the thoracic spine. Laterally directed anterior vertebral body screws were placed into each body and two vertebrae, either T6 and T7 or T8 and T9, were tethered with a stainless steel cable. After 12 weeks, radiographs were obtained to evaluate the degree of deformity that had developed. In addition, biomechanical testing to determine the range of motion in the tethered and untethered segments was performed. RESULTS: The calves increased their weight 153% during the 12-week postoperative period. The radiographic analysis demonstrated scoliosis of 11.6 degrees +/- 4.8 degrees in the tethered levels, as compared with 0.3 degrees +/- 1.7 degrees in the control segments (P < 0.0001). Similarly, kyphosis developed in the tethered segments (5.1 degrees +/- 5.8 degrees ), as compared with -1.8 degrees +/- 3.1 degrees at the control levels (P = 0.01). There was a significant wedging of the disc in the tethered (6.8 degrees +/- 1.6 degrees ) as compared with the untethered (0.7 degrees +/- 2 degrees ) segments (P < 0.0001). There was a trend toward lower height of the vertebrae on the tethered right side, as compared with the left side in the tethered segments (P = 0.075), whereas no side-to-side difference was noted in the control subjects (P = 0.48). Biomechanical analysis showed that the tether did not affect the range of motion in axial rotation or flexion-extension. However, the tether did restrict lateral bending as compared with that of the control subjects. When the tether was cut, the range of motion returned to levels matching that of the untethered control subjects. CONCLUSIONS: Anterolateral tethering of the spine creates kyphosis and scoliosis in this rapidly growing bovine model. The spinal tether limited motion primarily in lateral flexion. However, total lateral bending motion returned to levels comparable with control motion segments after removal of the tether. This method of spine growth modulation may provide a possible treatment for the correction of spine deformities without arthrodesis in patients who are skeletally immature. The exact mechanisms of growth modulation and the effects of tethering on disc function and integrity are unknown and deserve further study. PMID- 11923661 TI - Muscle spindle distribution, morphology, and density in longus colli and multifidus muscles of the cervical spine. AB - STUDY DESIGN: Tissue blocks comprising muscle and bone from C5 to C7 segments were harvested at autopsy from 16 individuals ranging in age from 4 to 77 years. The prevertebral longus colli and postvertebral multifidus muscle pairs from one side in each individual were randomly selected for this study of muscle spindles. OBJECTIVES: To determine muscle spindle distribution, morphology, and density for the longus colli and multifidus in caudal segments of the human cervical spine, and to assess whether changes are evident from infancy to old age. SUMMARY OF BACKGROUND DATA: Age-related changes to the osteoligamentous framework of the cervical spine have been well documented. Postural modification accompanies these structural alterations, but there have been limited attempts to document whether muscle sustains a comparable level of morphologic alteration. Previous studies have examined muscle spindles in the neck muscles of various animal models and in a variety of isolated human muscles. However, most of these studies incurred bias through sampling and methodologic assumptions. METHODS: The longus colli and multifidus were resected between C5 and C7, and between left and right pairs selected randomly for spindle analysis. These vertebral segments were selected deliberately because they form the apex of the cervical lordosis and the site at which the greatest age-related modification occurs. The tissue was processed in paraffin, sectioned, and then stained by Masson's trichrome. Spindle characteristics were examined using light microscopy and analyzed by unbiased stereologic methods. A one-sample paired t test was used to ascertain whether the differences in spindle density between the two muscles were statistically significant. RESULTS: The longus colli has a high density of muscle spindles, which appear clustered and concentrated anterolaterally, away from the vertebral body. The multifidus has a low density of muscle spindles, which are found predominantly as single units concentrated closely to the vertebral lamina. No change in spindle distribution, morphology, and density were observed with age. CONCLUSIONS: The current study examined spindle characteristics for an intrinsic neck muscle pair whose coactivation contributes to segmental stability of the cervical spine. The distribution and morphology of muscle spindles differ between the longus colli and the multifidus. In addition, these muscles have significant differences in terms of mean spindle density. Spindle characteristics represent one of many factors that govern proprioceptive regulation in skeletal muscle, and in neck muscles, the central connectivity of these receptors remains undefined. Therefore, although there are anatomic differences between the neck flexor and extensor, the functional implications of these differences are not clear. It is also of interest that spindle characteristics remain unchanged in these intrinsic muscles whose underlying segments are subject to age-related osteoligamentous changes. PMID- 11923662 TI - Biomechanical analysis of anterior scoliosis instrumentation: differences between single and dual rod systems with and without interbody structural support. AB - STUDY DESIGN: Nondestructive biomechanical testing was performed on bovine lumbar spines instrumented with multilevel scoliosis type anterior spine constructs. OBJECTIVE: To determine the biomechanical effects from the number of anterior rods (1 vs 2) and the effects of interbody structural support on construct stiffness after anterior multisegmental instrumentation. SUMMARY OF BACKGROUND DATA: Corrective surgery using anterior instrumentation for thoracolumbar and lumbar scoliosis has been performed with single rod and, more recently, with dual rod constructs. The biomechanical effect of one- or two-rod anterior instrumentation systems on construct stiffness and the addition or absence of interbody structural support have not been defined adequately in the literature. METHODS: Eight bovine lumbar spines each underwent instrumentation using four different constructs: one rod without interbody support; one rod with titanium mesh interbody support at the L2-L3, L3-L4, and L4-L5 disc spaces; two rods alone; and two rods with interbody support. Nondestructive cyclic testing in flexion-extension (+/-5 Nm), lateral bending (+/-5 Nm), and torsion (+/-2 Nm) were performed. The construct stiffness (Nm/ degrees ) of the four implant configurations was compared. RESULTS: With the addition of a second rod, the construct was significantly stiffer than a single rod construct in flexion (P = 0.006), extension (P = 0.02), and torsion (P = 0.01), but not in lateral bending. The addition of interbody structural support to the rod systems resulted in significantly stiffer constructs than those without cages in flexion (P = 0.03), but not in the other loading conditions (extension, lateral bending, torsion). CONCLUSIONS: Dual rod constructs were stiffer in torsion and flexion-extension loading than single rod constructs. Neither the number of rods nor the use of structural mesh interbody support had any effect on lateral bending stiffness. However, in a single rod system, the addition of interbody support increased stiffness in flexion. The use of structural support in dual rod constructs may be helpful in "setting" the desired lordosis, but adds little to construct stiffness. PMID- 11923663 TI - Biomechanical study and digital modeling of traction resistance in posterior thoracic implants. AB - STUDY DESIGN: Posterior implants were used to test the resistance of seven human thoracic spines to traction, and results were compared with those of a digital study. OBJECTIVES: To compare the use of hook and screw fixation methods for posterior thoracic surgery in a study supplemented by a digital analysis using the finite-element method. SUMMARY OF BACKGROUND DATA: The maximum break strength of pedicular screws on bone has already been studied several times. Far fewer studies have investigated the stresses that may be applied to pedicular and laminar hooks or compared these hooks with pedicular screws in the thorax. METHODS: For this study, seven human thoracic spines were used, identifying 49 groups of two vertebrae each. The assemblies used a bolt in the medullary cavity. The central diameter of this bolt could be varied using tubes to suit the individual medullary cavity, spreading the stresses over the entire centrum during traction. The assembly was placed on an Instron machine. Vertical traction was applied up to the maximum break strength, that is, the strength recorded at the top of the traction curve read out directly on the Instron machine. Traction speed in all the tests was 5 mm per minute. Four pedicular screws and two pedicular-laminar clips were used alternately. In these tests, 25 values were obtained with hooks and 24 with screws. The group with screw fixation was tested in two halves, with 12 tests using 4-mm diameter screws and 12 tests using 5-mm diameter screws. For the digital study, the vertebra model comprised nearly 63,000 nodes and 14,000 elements. Elastic field calculations were carried out using the Abaqus finite-element calculation method. RESULTS: Each time pedicular laminar traction was used, the pedicles broke at the base. Where screws were used, a medial fissure was found at the base of the pedicle. For the hooks, the maximum break strength was 1150 +/- 388 N. It was 820 +/- 418 N when 4-mm diameter screws were used and 1395 +/- 435 N when 5-mm screws were used. The most fragile vertebrae were found to be at T5-T6 and T7-T8. The screw-instrumented model showed that stresses were concentrated at the medial part of the pedicle, inside the medullary cavity. Use of a long screw did not lead to noticeable stress reduction. The hook-instrumented model showed that the stresses were greatest in the lower part of the pedicles. CONCLUSIONS: Screw fixation is the most appropriate from a mechanical point of view. During tests, however, screw fixation falls short of its promises. This leads to the hypothesis that in most cases screws pull out because the bony anchoring of the pedicle screwing fails. Screws are less effective if their anchoring in the pedicles cannot be guaranteed, which is probably the reason for their relative weakness. Screw diameter should be chosen according to the size of each pedicle. The findings showed that hooks impose additional stresses on the vertebrae. PMID- 11923664 TI - Microsurgical transdural discectomy with laminoplasty: New treatment for paracentral and paracentroforaminal cervical disc herniation associated with spinal canal stenosis. AB - STUDY DESIGN: A clinical study of the surgical procedure for cervical disc herniation was conducted. OBJECTIVES: To describe microsurgical transdural discectomy with laminoplasty, and to assess the clinical outcome of this surgical technique. SUMMARY OF BACKGROUND DATA: A posterior approach for cervical disc herniation has been considered risky, and few reports on a transdural approach to this disorder have appeared in the past decade. However, a transdural approach with recent innovations (a microsurgical technique, intraoperative spinal cord monitoring, and laminoplasty) has not been reported. METHODS: For this study, 30 patients with myelopathy or radiculomyelopathy accompanied by cervical disc herniation, aged 30 to 77 years (mean, 55 years), underwent microsurgical transdural discectomy with laminoplasty. Preoperative images showed multisegmental disc degeneration, developmental canal stenosis, or both for all the patients. The intraoperative evoked spinal cord potentials were recorded for neurophysiologic assessment. The follow-up period averaged 52 months (range, 24 118 months). RESULTS: The operative time averaged 239 minutes (range, 160-340 minutes), and the mean blood loss was 169 mL (range, 30-701 mL). The Japanese Orthopedic Association score improved from 3.5 to 15 (mean, 11.4) before surgery to 9 to 17 (mean, 15.2) after surgery. The intraoperative evoked spinal cord potentials indicated the affected spinal cord level and reflected the severity of myelopathy. Postoperative cerebrospinal fluid leakage, pseudomeningocele, and progression to cervical deformity were not observed. Transient palsy of the C5 nerve root was observed in two patients with C4-C5 central cervical disc herniation. CONCLUSIONS: Microsurgical transdural discectomy with laminoplasty can be performed safely as a selected surgical option for paracentral and paracentroforaminal cervical disc herniation with multisegmental canal stenosis. PMID- 11923665 TI - Posterolateral endoscopic excision for lumbar disc herniation: Surgical technique, outcome, and complications in 307 consecutive cases. AB - STUDY DESIGN: A retrospective review involving 307 consecutive cases of lumbar disc herniation managed by posterolateral endoscopic discectomy was conducted. OBJECTIVES: To describe a contemporary posterolateral endoscopic decompression technique for radiculopathy secondary to lumbar disc herniation; to evaluate the efficacy of the technique as it is applied to lumbar disc herniation including primary herniation, reherniation, intracanal herniation, and extracanal herniation; and to report outcome and complications. SUMMARY OF BACKGROUND DATA: The concept of percutaneous posterolateral nucleotomy was introduced in 1973. The development of the related equipment and technique had witnessed a slow and lengthy evolution. METHOD: A retrospective assessment of 307 patients was performed at least 1 year after their index operation. The outcome was graded according to a modified MacNab method. A patient-based outcome questionnaire also was incorporated into the study. RESULTS: The surgeon-performed assessment showed satisfactory results in 89.3% of the cases. The rate of response to the questionnaire was 91%. The responses indicated that 90.7% of the respondents were satisfied with their surgical outcome and would undergo the same endoscopic procedure again if faced with a similar herniation in the future. The poor outcome occurred in 10.7% of the primary group and 9.7% of the questionnaire group. The combined major and minor complication rate was 3.5%. CONCLUSIONS: The surgical outcome of posterolateral endoscopic discectomy for lumbar disc herniation is comparable with that for the traditional open transcanal microdiscectomy. Intracanal and extracanal herniations, reherniations, and incidental lateral recess stenosis can be addressed by the same approach. PMID- 11923666 TI - Analysis of chondrex (YKL-40, HC gp-39) in the cerebrospinal fluid of patients with spine disease. AB - STUDY DESIGN: The expression of chondrex (YKL-40, HC gp-39) was measured in the cerebrospinal fluid of from patients with spine diseases. OBJECTIVES: To quantify the levels of chondrex in human cerebrospinal fluid, and to clarify the nature of its expression. SUMMARY OF BACKGROUND DATA: Chondrex is a newly discovered 40-kDa glycoprotein identified originally in the whey secretions of nonlactating cows. It is secreted by a human osteosarcoma cell line, human articular cartilage chondrocytes, and human fibroblasts. However, the function of chondrex in chondrogenesis is unknown, and the expression of chondrex in human cerebrospinal fluid has never been reported. METHODS: The concentration of chondrex in human cerebrospinal fluid was measured by sandwich immunoassay with antihuman chondrex antibodies. Cerebrospinal fluid samples were collected from two groups of patients. Group 1, the control group, consisted of 34 trauma patients. Group 2 consisted of 130 patients with spine diseases: 29 with cervical spondylotic myelopathy, 30 with lumbar disc herniation, 35 with lumbar canal stenosis, and 36 with scoliosis. All values are expressed as the mean +/- standard deviation. RESULTS: The concentration of chondrex in Group 1 (control group) was 113.8 +/- 48.3 ng/mL. The concentrations of chondrex in Group 2 were 245.3 +/- 107.2 ng/mL in cervical myelopathy, 143.2 +/- 53.6 ng/mL in lumbar disc herniation, 241.5 +/- 77.2 ng/mL in lumbar canal stenosis, and 71.4 +/- 33.9 ng/mL in scoliosis. The concentrations of chondrex in cervical myelopathy, lumbar canal stenosis, and lumbar disc herniation were significantly higher than in the control group (P < 0.05). CONCLUSIONS: In this study, the chondrex concentration was high in spine diseases causing spinal stenosis. The authors believe that chondrex is expressed in cerebrospinal fluid as a result of damage or stress to the neural structure, and that it could be a new marker for spine diseases. PMID- 11923667 TI - Cochrane review on the role of surgery in cervical spondylotic radiculomyelopathy. AB - STUDY DESIGN: This study involved a search of MEDLINE (1966 to 2000), EMBASE (1980 to 2000), and the Cochrane Controlled Trials Register. The authors of the identified randomized controlled trials were contacted to detect any additional published or unpublished data. The trials selected for this study included all the truly unconfounded or quasi-randomized controlled investigations allocating patients with cervical radiculopathy or myelopathy to 1) "best medical management" or "decompressive surgery (with or without some form of fusion) plus best medical management," or 2) "early decompressive surgery" or "delayed decompressive surgery." Two reviewers independently selected trials for inclusion, assessed trial quality, and extracted the data. OBJECTIVES: To determine whether surgical treatment of cervical radiculopathy or myelopathy is associated with improved outcome, as compared with conservative management, and whether the timing of surgery (immediate or delayed because of persistence or progression of relevant symptoms and signs) has an impact on outcome. SUMMARY OF BACKGROUND DATA: Cervical spondylosis causes pain and disability by compressing the spinal cord or roots. Surgery to relieve the compression may reduce the pain and disability. However, it is associated with a small but definite risk. This study sought to assess the balance of risk and benefit from surgery. METHODS: Two trials involving a total of 130 patients were included. One trial with 81 patients compared surgical decompression with either physiotherapy or cervical collar immobilization in patients with cervical radiculopathy. RESULTS: The short term effects of surgery, in terms of pain, weakness, or sensory loss were superior. However, at 1 year no significant differences between the groups were observed. Another trial with 49 patients compared the effects of surgery with those of conservative treatment in patients who had a mild functional deficit associated with cervical myelopathy. No significant differences were observed between the groups up to 2 years after treatment. CONCLUSIONS: The data from the reviewed trials were inadequate to provide reliable conclusions on the balance of risk and benefit from cervical spine surgery for spondylotic radiculopathy or myelopathy. PMID- 11923668 TI - Ten-year follow-up study of lower thoracic hemivertebrae treated by convex fusion and concave distraction. AB - STUDY DESIGN: A retrospective review of patient records with recent clinical and radiologic assessment was conducted. OBJECTIVE: To evaluate the long-term results of fully segmented hemivertebrae treated by convex fusion combined with instrumented concave subcutaneous distraction. SUMMARY OF BACKGROUND DATA: Convex fusion has been described for the treatment of hemivertebrae in children, whereas distraction without fusion has been shown to enhance spinal growth. No long-term follow-up studies have combined these two methods. METHODS: Between 1986 and 1994, six consecutive patients (5 males and 1 female) with hemivertebrae located at T11 or T12 underwent convex anterior and posterior fusion as well as concave subcutaneous distraction without fusion. RESULTS: The mean age at surgery was 3.4 years. The mean follow-up period was 10.8 years (range, 8-14 years). There was a mean improvement of 41% in the coronal deformity, from a mean angle of 49 degrees before surgery to 29 degrees at the latest follow-up assessment. In four of the cases, this correction was achieved immediately after surgery and did not significantly change despite repeated distraction. The kyphosis improved in three cases, remained unchanged in one case, and deteriorated in two cases. In these two cases, an adjacent wedge vertebra contributed to the kyphotic deformity. CONCLUSIONS: Although growth-mediated correction was seen in only two cases, this procedure could be recommended for children with severe deformities and decompensation in the lower thoracic spine. It is safer than hemivertebra excision, with less risk of spinal cord injury. The concave distraction produces immediate improvement in the coronal balance, such that there is no need to wait for uncertain growth-mediated correction in patients who undergo convex fusion only. PMID- 11923670 TI - Point of view. PMID- 11923669 TI - Sagittal plane correction in idiopathic scoliosis. AB - STUDY DESIGN: Patients with idiopathic scoliosis who had undergone posterior fusion by means of posterior multisegmented hook instrumentation were studied retrospectively. OBJECTIVES: To present the changes in projected thoracic hypokyphosis and the behavior of lumbar lordosis within and below the fusion. SUMMARY OF BACKGROUND DATA: Scoliosis is a three-dimensional deformity of the spine. The idiopathic cases usually exhibit a flattening of the sagittal curves, which had further deteriorated when the Harrington technique was used. The consequences included the flat back, angular increase of the lumbar lordosis below the fusion, and low back pain. Previous studies showed no or only moderate correction of thoracic hypokyphosis when using Cotrel-Dubousset instrumentation or its modifications were used. Harrington rod systems resulted in decreased lumbar lordosis in the fusion area and increased lordosis below the fusion. No background data were found concerning the effects of multisegmented hook instrumentation on the lumbar spine within and below the fusion. METHODS: For this study, 306 patients with idiopathic scoliosis who had undergone posterior spinal fusion with multisegmented hook systems using the derotation maneuver were analyzed after a mean follow-up period of 5 years and 4 months. The coronal plane curvature, the sagittal plane projection of the thoracic kyphosis, and the lumbar lordosis within and below the fusion were evaluated. RESULTS: The average coronal plane correction was 67.1%. Analysis of the sagittal contours demonstrated that the preoperative thoracic hypokyphosis (less than 20 degrees between T4 and T12) increased by an average of 12 degrees, and that 55.1% of hypokyphotic backs were corrected to the normal range (20 degrees to 40 degrees ). In patients with frank lordosis (kyphosis less than 10 degrees ), the degree of correction was higher (average, 16 degrees ), but complete correction was achieved in only 38.5% of the cases. In patients with mild lordosis (kyphosis between 10 degrees and 20 degrees ), the average correction was 8 degrees, and 71.3% of the patients were in the normal range after surgery. The normal preoperative thoracic kyphosis was preserved in 81.3% of the cases. In the lumbar area, the Cotrel-Dubousset instrumentation was capable of correcting the preoperative hypolordosis (less than -20 degrees between L1 and L5) in 94.4% of the cases. The normal preoperative lordosis (-20 degrees to -60 degrees ) was preserved in 97.9% of the cases. The hyperlordosis was corrected in all cases. Analysis of the data in terms of lower fusion limit showed that the lower the caudal hook, the greater the increase in the segmental lordosis within the fusion, without any increase distal to the fusion. No segmental hyperlordosis was observed below the fusion. CONCLUSIONS: The Cotrel-Dubousset technique ensures considerable sagittal correction of the spine. In the course of scoliosis correction, it is possible to preserve the normal preoperative sagittal profile of the spine, to correct the slightly flattened thoracic kyphosis, to increase materially the kyphosis of the frankly hypokyphotic spine, to preserve or restore normal lumbar lordosis in a considerable percentage of the cases, to avoid angular segmental hyperlordosis at the level of the first disc below the fusion, and to avoid retrolisthesis of the last fused vertebra. PMID- 11923671 TI - Multisurgeon assessment of coronal pattern classification systems for adolescent idiopathic scoliosis: reliability and error analysis. AB - STUDY DESIGN: Three scoliosis surgeons and one orthopedic fellow were presented the anteroposterior radiographs of 70 patients with adolescent idiopathic scoliosis. All the reviewers assigned a type to each curve according to the classification systems of H. A. King and R. W. Coonrad. OBJECTIVES: To compare multisurgeon reliability in applying the classification systems of H. A. King and R. W. Coonrad, and to analyze controversially classified curve patterns. SUMMARY OF BACKGROUND DATA: The system most commonly used to classify adolescent idiopathic scoliosis is King's classification. However, because of poor interobserver reliability, the validity of this system is questioned. In contrast, high interobserver reliability is reported for Coonrad's classification system, which is used less frequently in clinical practice. METHODS: Interobserver agreement and intraobserver reproducibility were tested. Kappa coefficients were used to test reliability. Between the observers, the divergent assignments to curve patterns were analyzed in both quantitative and qualitative terms. An error analysis was performed. RESULTS: Paired comparisons showed a mean interobserver kappa coefficient of 0.45 for King's and 0.38 for Coonrad's classification systems. According to Svanholm et al, these values indicate poor reliability in terms of interobserver agreement. Error analyses for both classification systems showed that the reason for poor reproducibility is disagreement among the observers about structural upper thoracic and structural lumbar curves. CONCLUSIONS: Neither the King nor the Coonrad method appears to have sufficient interobserver reliability. To improve reliability, the authors recommend that the structural stigmas of the upper thoracic and lumbar curves be unequivocally described. PMID- 11923672 TI - Platelet calmodulin levels in adolescent idiopathic scoliosis: do the levels correlate with curve progression and severity? AB - STUDY DESIGN: This ongoing longitudinal study evaluates simultaneous radiographic and platelet calmodulin determinations for patients with idiopathic scoliosis who are skeletally immature. OBJECTIVES: To determine whether platelet calmodulin levels correlate with curve progression and severity. SUMMARY OF BACKGROUND DATA: A previous study based on a single calmodulin determination and a single radiograph identified higher calmodulin levels in progressive curves and in higher magnitude curves. A longitudinal study was needed to demonstrate the relation of calmodulin to curve changes for individual patients over time during the growth period. METHODS: In this study, 55 patients with idiopathic scoliosis of varying types and severity were followed longitudinally with serial radiographs and platelet calmodulin determinations. A Risser sign was recorded for each radiograph at each visit. RESULTS: Calmodulin levels increased in all the patients with progressive curves (13/13), remained stable in 73% of the patients with nonprogressive curves (11/15), and were higher generally in curves greater than 30 degrees and double structural curves. Calmodulin levels usually decreased in patients undergoing brace treatment (14/17) or spine fusion (9/10). CONCLUSIONS: It appears that platelet calmodulin levels correlate closely with curve progression and stabilization by bracing or spine fusion. Correlation with nonprogressive curves was not as consistent, with 27% noncorrelation. Longer follow-up evaluation and enrollment of additional patients will be necessary to determine whether calmodulin may serve as a biochemical marker of curve progression and to help identify stable and progressive curves. PMID- 11923673 TI - Outcome and complications of long fusions to the sacrum in adult spine deformity: luque-galveston, combined iliac and sacral screws, and sacral fixation. AB - STUDY DESIGN: A retrospective study of adults with long fusion to the sacrum using three different fixations was performed. OBJECTIVE: To compare the long term clinical results and complications associated with three methods of lumbosacral fixation for adult spine deformities: Luque-Galveston, combined iliac and sacral screws, and sacral screws. SUMMARY OF BACKGROUND DATA: The preferred technique for long fusion to the sacrum is controversial, and surgery for adult deformity is fraught with significant technical difficulties and high complication rates. No clinical study compares the long-term outcome of long fusion to the sacrum using these different methods of lumbosacral fixation. METHODS: This study included 54 consecutive patients who underwent elective combined anterior and posterior surgical reconstruction for adult spine deformity with a minimum follow-up period of 2 years. The patients were divided into three groups on the basis of the surgical method used for the posterior spine instrumentation. Group 1 consisted of 11 patients with smooth L-rod and segmental sublaminar wire instrumentation (Luque-Galveston technique). Group 2 consisted of 36 patients with posterior Isola segmental instrumentation and combined iliac and sacral screws. Group 3 consisted of 12 patients with Isola segmental instrumentation using bicortical sacral screws. Five patients were revised to another fixation group, giving a total of 59 cases. Radiographic, clinical results, and long-term outcome data were obtained using the modified Scoliosis Research Society (SRS) outcome instrument. RESULTS: There were 26 late complications. Pseudarthrosis developed in 10 patients, requiring revision surgery: 4 (36%) in the Group 1, 5 (14%) in Group 2, and 1 (8.5%) in Group 3. Comparison of the modified SRS outcomes showed no difference among the groups. The average SRS grand total score was 73.4% for Group 1, 70.9% for Group 2, and 62.6% for Group 3. Overall, 76% of the patients were satisfied with their outcome. The presence of perioperative complications or pseudarthrosis significantly correlated with a lower satisfaction score (P = 0.012 and P = 0.048, respectively). Sagittal plane decompensation significantly correlated with a higher pain score (P = 0.035). Patients with prior surgeries scored lower on the self-image questions than patients with no prior surgery (P = 0.007). CONCLUSIONS: Attention to sagittal balance is critical in these patients. Revision surgery is as safe and effective as primary surgery. According to the current findings, the Luque-Galveston fixation technique has an unacceptably high rate of pseudarthrosis, and this method is not recommended for adult deformities. Currently, the authors are using bicortical and triangulated sacral screws with an anterior interbody support in patients with good bone stock, but only when the spine balance is restored. Otherwise, they recommend using iliac fixation, although there is a higher rate of painful hardware, requiring removal. PMID- 11923674 TI - Dorsal root entry zone lesioning used to treat central neuropathic pain in patients with traumatic spinal cord injury: a systematic review. AB - STUDY DESIGN: A systematic review was conducted for studies of any design. OBJECTIVES: To evaluate the effectiveness and safety of dorsal root entry zone lesioning in treating central neuropathic pain of patients with traumatic spinal cord injury. SUMMARY OF BACKGROUND DATA: Central neuropathic pain has a great impact on the quality of life of many patients with traumatic spinal cord injury. Dorsal root entry zone lesioning has become one of several treatment options. No previous systematic reviews were found that address this indication for the procedure. METHODS: Several computerized databases were searched along with reference lists of eligible articles and personal files of advisory panel members. Only primary studies of patients older than 13 years with both traumatic spinal cord injury and central neuropathic pain receiving dorsal root entry zone lesioning were included. Two assessors independently performed data extraction and quality assessment for all the studies. RESULTS: The selection criteria were met by 11 studies. All were case series of 5 to 54 patients. As reported by 10 of the studies, at least 50% of the patients attained more than 50% pain relief or experienced no pain limitation of activity and no need for narcotics. However, all the studies had poorly defined eligibility criteria or none at all, no control groups, and inadequate reporting of adverse effects. Only one study provided adequate patient description. None of the studies reported the validity, reliability, or responsiveness of outcome measures. CONCLUSIONS: Reviewed studies indicate promising results from dorsal root entry zone lesioning for the treatment of central neuropathic pain in selected patients with traumatic spinal cord injury. However, the strength of the evidence provided by the studies was poor in terms of study design, outcome measures, reports on the severity of adverse effects, patient selection criteria, and patient description. For these reasons, the evidence is weak for the use of dorsal root entry zone lesioning to relieve central neuropathic pain in patients with traumatic spinal cord injury. PMID- 11923675 TI - Patient education on the internet: opportunities and pitfalls. AB - The Internet, an increasingly pervasive part of our culture, has vastly increased the rate at which information is disseminated. Unfortunately, a substantial portion of medical information on the Internet is false or misleading. Realizing this, many patients would like their physicians to help guide their online searches for medical information, but most physicians do not feel comfortable enough with the Internet to provide this service. A recent poll by Cyberdialogue (2000) found that 70% of all patients would like their physicians to recommend a health care web site for their condition, but that only 4% of patients receive such a recommendation. In recommending medical web sites, physicians need to consider the quality of the web site, the quality of the content, and especially the ethics of the site. The most important ethical standard a web site must meet is the clear separation of its editorial comment from promotional material. Patients do not have enough knowledge to distinguish unbiased information from information designed to push a product or service. There are organizations such as the Health on the Net Foundation and the E-Health Code of Ethics that have established clear quality and ethical criteria, but very few resources exist for these organizations to use in policing the sites that claim to adhere to their criteria. The National Institutes of Health recently started a consumer web site (www.medlineplus.gov) that takes web sites that adhere to their stringent criteria and places them on their web site, thus ensuring proper compliance. Patient education from sources other than face-to-face contact with a physician should be seen as an adjunct to the medical practice of physicians, and not as competition. Information is a form of therapy, and should lead to better referrals, more realistic outcome expectations, better treatment compliance, and better outcomes. As long as patients are referred to information that is unbiased, well-written, and not proprietary, their decision making should be enhanced, bringing them into the decision-making loop. This allows the patient to be much more involved with his or her care. As the medical profession changes from a "gatekeeper" model to a patient choice model, patient education will take on a much greater role. PMID- 11923676 TI - Primary epidural hemangiopericytoma in the lumbar spine: a case report. AB - STUDY DESIGN: A case report of primary epidural hemangiopericytoma in the lumbar spine and a review of the literature are presented. OBJECTIVE: To present the result of pathologic diagnosis using immunohistochemical staining and the treatment of spinal hemangiopericytoma. SUMMARY OF BACKGROUND DATA: Spinal hemangiopericytoma is a very rare soft tissue tumor with specific pathologic features and a clinical course featuring high rates of recurrence and metastasis. METHODS: A 39-year-old woman reported numbness in both legs. Neither sensory abnormalities nor muscle weakness was present in her lower extremities. Magnetic resonance imaging showed a tumor dorsal to the thecal sac at L1-L2. After L1 and L2 laminectomy, the tumor with its dural base was resected en bloc. RESULTS: The patient's clinical and neurologic symptoms disappeared after surgery. Microscopic examination showed oval- or spindle-shaped cells with slightly acidic cytoplasm and oval nuclei. Silver staining emphasized fibers around tumor cells. The test results for the tumor cells were positive for vimentin staining, but negative for alpha-TM staining using thrombomodulin, a marker for endothelial cells. On the basis of these pathologic findings, the tumor was diagnosed as a hemangiopericytoma, a type of tumor composed of mesenchymal hemangiopericytes. Neither recurrence nor metastasis of the tumor was found during the 2-year follow up period after surgery. CONCLUSIONS: Soft tissue hemangiopericytoma is a well recognized entity considered to be an aggressive neoplasm with a high rate of recurrence and a propensity to metastasize. Immunohistochemical investigation was essential for the diagnosis of this tumor. Although hemangiopericytoma very rarely occurs in the spine, surgeons treating patients with this tumor should be aware of its metastatic potential. PMID- 11923677 TI - Dumbbell-shaped meningioma with cystic degeneration in the thoracic spine: a case report. AB - STUDY DESIGN: A case report of dumbbell-shaped meningioma in the thoracic spine with cystic degeneration for which a combined posterior microscopic and anterior thoracoscopic approach was used is described. OBJECTIVES: To report a rare case of dumbbell-shaped meningioma, and to describe a thoracoscopic surgical approach for paravertebral tumor. SUMMARY OF BACKGROUND DATA: Dumbbell-shaped tumors arising from nerve root sheaths are very common. However, a dumbbell-shaped meningioma is relatively rare, and reports on thoracoscopic resection of dumbbell and paravertebral tumors are still very few in number. METHODS: The dumbbell meningioma was managed surgically. Because of the paravertebral tumor's intrathoracic expansion, a combined posterior and anterior approach was used. With the posterior approach, microscopic resection using hemilaminotomy was performed, whereas with the anterior approach, the thoracoscopic approach was used instead of thoracotomy. RESULTS: Complete resection of the tumor was performed successfully. The patient regained his functional ability to walk soon after surgery because of less pain than with thoracotomy. CONCLUSIONS: With combined microscopic and thoracoscopic surgery, dumbbell-shaped meningioma could be completely resected. The thoracoscopic approach for the removal of a paravertebral tumor can be a good alternative to thoracotomy. PMID- 11923678 TI - Combined odontoid and jefferson fracture in a child: a case report. AB - STUDY DESIGN: A case of combined odontoid and Jefferson fracture is reported. OBJECTIVE: To alert spine physicians to the rare combination of an odontoid and Jefferson fracture in a child. METHODS: A 5-year old boy presented with neck pain and torticollis after falling on his head from a four-wheeler that had rolled over. A computed tomography scan confirmed a combined odontoid and Jefferson fracture. RESULTS: The child was successfully treated nonsurgically with a hard cervical orthosis. At this writing, the child clinically is asymptomatic 2 years after the injury. DISCUSSION: The fall on to the head caused the body weight to be transmitted to the atlas. The resulting force vector produced the classic Jefferson fracture of the atlas. As the atlas fracture spread with continued compressive and axial forces, tension was exerted on the alar ligaments (check ligaments), leading to the avulsion fracture of the odontoid. CONCLUSIONS: This is only the second reported case of a child with a combined Jefferson and odontoid fracture. This diagnosis should be considered in the evaluation of a child with neck pain and torticollis from a fall on the top of the head. PMID- 11923679 TI - Logistical problems have a tendency to hinder cornea procurement. PMID- 11923680 TI - The preclinical model of choice. PMID- 11923681 TI - Cyclosporin monitoring based on C2 sampling. PMID- 11923682 TI - Intrathymic islet transplantation in the canine: I. Histological and functional evidence of autologous intrathymic islet engraftment and survival in pancreatectomized recipients. AB - BACKGROUND: Although an attractive alternative to daily insulin therapy, allogeneic pancreatic islet transplantation has yielded suboptimal results in clinical trials, in contrast to islet allotransplantation in animal models, which have demonstrated consistent success. The successful transplantation of isolated islets to the thymus, with a single concomitant dose of antilymphocyte serum, has been demonstrated in rodents, and more significantly, such intrathymic islet allografts have been shown to induce recipient tolerance toward subsequent extrathymic donor strain islet allografts. Intrathymic islet autotransplantation has been pursued, as a prelude to studies of allogeneic IT islet transplantation and tolerance induction, in canine, porcine, and non-human primate models, to assess the large animal thymus as a site capable of supporting a viable islet graft. However, little functional or histological evidence has established definitive survival of islets transplanted within the thymus of a phylogenetically advanced species, which may be requisite to tolerance induction. This study describes the successful intrathymic autotransplantation of isolated islets using a canine model. METHODS: Purpose-bred juvenile dogs, aged 4-6 months, underwent partial (n=4), or total pancreatectomy (n=11), and transplantation of autologous islets. The pancreas (or pancreatic limb) was distended with collagenase solution, and digested using a modification of the semiautomated system of Ricordi. Islets were purified by discontinuous gradient centrifugation, using Euroficoll (ficoll in Euro-Collin's kidney preservation solution). Partially pancreatectomized canines underwent IT transplantation of purified autologous islets (8000+/-4000 IEs), and were killed 8 weeks posttransplant. Totally pancreatectomized canines underwent transplantation of autologous islets to the liver (via portal vein embolization, n=5, IPO group) or the thymus (via direct IT injection, n=6, IT group), and were serially evaluated for a period of 8 weeks posttransplant to assess fasting blood glucose (FBG), serum insulin (SI) levels, and i.v. glucose tolerance (IVGTTs). K values (defined as the %-decrease/minute of the log(e) of blood glucose values) were calculated from IVGTT results. RESULTS: After autotransplantation in this cohort of animals, five of five IPO, and three of six IT islet recipients, remained normoglycemic (mean FBG< or =250 mg%) immediately posttransplant, and all recipients exhibited significantly elevated SI levels compared to apancreatic controls (n=10, followed 72 hr postpancreatectomy). Normal k values (=-1.1) were observed in two of five IPO, and in one of six IT recipients, 8 weeks after transplantation, and thymic tissue insulin content was increased compared to non-islet-bearing thymi (93.7+/ 48.6 ng/g tissue vs. 0.7+/-0.4 ng/g tissue). At 8 weeks posttransplantation thymi from both partially and totally pancreatectomized animals were resected and processed for histological examination. Microscopic analysis of islet-bearing thymi revealed positive staining for islet-specific hormones (insulin and glucagon) within all IT recipients., Identification of islets within thymi of hyperglycemic IT recipients was problematic as islet beta cells were highly degranulated as a result of the recipients glycemic state. CONCLUSIONS: These results indicate that autologous islets, transplanted to the canine thymus, engraft, function, and survive for up to 8 weeks after islet autotransplantation to the canine thymus and establish the feasibility of intrathymic islet transplantation in a phylogenetically advanced animal model. The ability of islets to survive within the thymic environment for a period of at least 8 weeks after transplantation suggests that the successful induction of specific unresponsiveness secondary to intrathymic transplantation will not be impaired or limited by the inability of a viable islet mass to survive within the thymus for a sufficient period. PMID- 11923683 TI - Analysis of robust innate immune response after transplantation in the absence of adaptive immunity. AB - BACKGROUND: Both animal models and clinical outcomes studies of transplantation suggest that antigen-independent mechanisms can alter graft survival and function. It has been suggested that antigen-independent processes interact with alloantigen-specific responses to augment the rejection reaction. A major link between antigen-specific adaptive immunity and pro-inflammatory stimuli is innate immunity. During transplantation, innate immunity may be stimulated by multiple factors, including ischemia, reperfusion, sterile injury, systemic stress, and cell death. METHODS: We investigated the hypothesis that transplantation induces a potent innate immune response in a murine model of vascularized solid organ transplantation. In our studies, we analyzed three experimental groups: (a) alymphoid group in which both the donor and recipients strains lacked an adaptive immune response due to deletion of the recombinase activating gene, thus blocking production of both T cell and B cell antigen receptors; (b) syngeneic group in which the donors and recipients were genetically identical; and (c) allogeneic group in which the donors and recipients had a complete MHC mismatch. To analyze a large number of parameters we determined the level of expression of a panel of cytokines, chemokines, receptors, and cell surface markers by RNase protection assays. In addition, serum cytokines were determined by ELISA and the infiltration of inflammatory cells was assessed by histology. RESULTS: Our results showed macrophage infiltration and up-regulation of multiple cytokines, chemokines, and chemokine receptors within the first day after transplantation in all groups, including the syngeneic and alymphoid recipients. CONCLUSIONS: Our study demonstrated a robust innate immune response that is independent of adaptive immunity and natural killer cell responses. PMID- 11923685 TI - Effects on T-cell maturation and proliferation induced by lactational transfer of cyclosporine to nursing pups. AB - BACKGROUND: Pregnancy after allotransplantations is becoming a more common occurrence, and the immunosuppressant of choice is cyclosporine (CsA) for these patients. Consequently, the effect of CsA on prenatal and postnatal immune development and function in the infant is an increasingly important clinical issue. The purpose of this study was to evaluate the potential problems of maternal CsA exposure on neonatal T-cell maturation and proliferation after lactational transfer of CsA in an animal model. METHODS: CsA was administered daily (subcutaneous) for 20 days during lactation, beginning the day of parturition using two dose levels (15 and 25 mg/kg body weight/day) in conjunction with saline controls. RESULTS: Considerable amounts of CsA were passed to the newborn rats with neonatal blood levels equal to that of the mothers for the 25-mg/kg/day dose and 55% for the 15 mg/kg dose. There was a significant reduction in thymus/body-weight ratio and thymus cellularity for the pups born to mothers dosed at 15 or 25 mg/kg/day of CsA. The thymus from the CsA exposed pups showed an almost complete loss of the medullary region with no apparent change in the thymic cortex. The CsA-treated mothers and their pups (15 and 25 mg/kg/day dose) had a significant increase in the percentage of CD4+CD8+ thymocytes and a significant decrease in the percentages of CD4+, CD3hi, and T cell receptor (TCR)hi thymocyte phenotype subsets and CD4/CD8 ratios. Thymocyte proliferative responses to concanavalin A + interleukin-2 were also significantly decreased in the mother and pup after both doses of CsA. In contrast to the mothers that showed no change in splenocyte proliferative responses, their pups showed decreased responses at both the 15- and 25-mg/kg doses. All immune alterations due to CsA lactational exposure in the pups were back to control levels after 30 days of postweaning CsA cessation. CONCLUSIONS: This study clearly demonstrates that neonatal exposure to CsA via lactational transfer can cause significant alterations in T-cell maturation and inhibition of lymphoproliferative responsiveness to mitogen activation. Although the CsA blood level in human transplant patients is normally much lower, this data indicate a potential for increased risk to opportunistic infections due to altered immune components in babies exposed to long-term CsA. PMID- 11923684 TI - Blockade of CD40-CD154 at the time of donor-specific blood transfusion does not lead to prolonged kidney allograft survival in nonhuman primates. AB - BACKGROUND: In rodents it has been demonstrated that blockade of the CD40-CD154 (CD40L) pathway at the time of donor-specific blood transfusion (DST) can result in indefinite graft survival. Because it has been reported in the past that DST in monkeys can have a favorable effect on graft outcome and that blockade of the CD40-CD154 pathway can lead to prolonged kidney graft survival in monkeys, we have combined anti-CD154 treatment with DST in a monkey kidney graft model. The aim of this study was to investigate the immunosuppressive potential of blocking the CD40-CD154 interaction at the time of a DST in rhesus monkeys. METHODS: One donor-derived blood transfusion was given on day -15 after the first anti-CD154 injection. The anti-CD154 antibody was given on days -15, -13, -11, -9, and -7. The kidney was transplanted on day 0. Cyclosporine was given after kidney trans plantation. RESULTS: No major difference in graft survival was observed between the groups. The animals died due to grade II acute rejection. At the time of transplantation, no antibody response could be detected directed against donor antigens. After transplantation, all animals surviving for more than 3 weeks had antidonor antibodies. There were no differences in the intragraft events analyzed by real time reverse transcriptase-polymerase chain reaction. CONCLUSIONS: DST under the cover of relatively high levels of anti-CD154 failed to result in prolonged graft survival or prevent the formation of antidonor antibodies, when cyclosporine was given after transplantation. PMID- 11923686 TI - Long-term kidney graft survival by delayed T cell ablative treatment in rhesus monkeys. AB - BACKGROUND: Tolerance to organ allografts in primates including man has been elusive, although in rodents and pigs tolerance can be achieved to organ allografts with relatively short courses of immunosuppressive treatment. In all varieties of graft acceptance that do not require full-dose maintenance immunosuppression, immunological engagement of donor and recipient and an early unstable period have been observed. On the basis of the hypothesis that elimination of aggressive T cell function should tip the balance in favor of an operationally tolerant state, experiments have been performed in monkeys allowing recipient-donor interaction before T-cell ablation and a short course of immunosuppression. METHODS: Rhesus monkeys received an allogeneic kidney graft from a MHC-mismatched donor. The animals either received anti-CD3 immunotoxin (FN18-CRM9) alone, started 2 days after transplantation, or in combination with a short course of cyclosporine (CsA) and/or rapamycin (RAPA), started at 5 days after transplantation. Kidney function was followed by monitoring serum creatinine levels and regular biopsies. Humoral and cellular antidonor immunity was tested in vitro before and at several time points after transplantation. RESULTS: Graft survival of monkeys that received CsA alone (mean survival time (MST)=29.3) was significantly prolonged compared with the controls (MST=6). FN18 CRM9 treatment alone also resulted in prolonged graft survival (MST=29.4). The combined treatment of FN18-CRM9 and CsA and/or RAPA resulted in prolonged graft survival after all immunosuppression was stopped (MST=207.8). CONCLUSIONS: It seems feasible to postpone immunosuppression posttransplantation and yet prevent allograft rejection without the need of permanent immunosuppression. PMID- 11923687 TI - Serum anti-pig antibodies as potential indicators of acute humoral xenograft rejection in pig-to-cynomolgus monkey kidney transplantation. AB - BACKGROUND: Hyperacute rejection of solid organ pig xenografts in nonhuman primates has been overcome by using donors transgenic for human complement regulatory proteins, but grafts are still susceptible to humoral (antibody mediated) rejection. We investigated whether circulating xenoreactive antibodies are a useful indicator of this xenograft rejection. METHODS: Five assays were employed in a retrospective analysis on 20 selected cynomolgus monkey recipients of renal xenografts transgenic for human decay-accelerating factor, with survival between 4 and 60 days. The assays included hemolytic and hemagglutination assays and the measurement of immunoglobulin (Ig)G and IgM binding to porcine endothelial cells and leukocytes, and to the Gal alpha 1-3Gal trisaccharide (Gal) antigen. To assess non-Gal-directed antibodies, sera were absorbed with a Gal coated resin. A predictive value was defined as an increase in antibody levels before a decline in graft failure (>20% increase in creatinine levels) and humoral rejection in graft pathology. RESULTS: Data on hemolytic anti-pig antibody correlated with those on IgM antibody to endothelial cells, leukocytes, and Gal. In absorbed sera IgM and IgG antibody to endothelial cells and leukocytes correlated with each other, indicative for an elicited antibody response to non-Gal antigens. Sixteen animals showed humoral rejection, and in all but two animals one or more assays was considered of predictive value. On the other hand, increased antibody levels were noted in two animals without signs of rejection in graft pathology and in two cases with cellular xenograft rejection. CONCLUSIONS: It is recommended to use multiple assays (preferably hemolytic, anti Gal, and anti-endothelial cell) to be able to fully monitor the peripheral antibody responses in pig-to-primate xenograft recipients. PMID- 11923688 TI - Overcoming severe renal ischemia: the role of ex vivo warm perfusion. AB - BACKGROUND: The ability to effectively utilize kidneys damaged by severe (2 hr) warm ischemia (WI) could provide increased numbers of kidneys for transplantation. The present study was designed to examine the effect of restoring renal metabolism after severe WI insult during ex vivo warm perfusion using an acellular technology. After warm perfusion for 18 hr, kidneys were reimplanted and evaluated for graft function. METHODS: Using a canine autotransplant model, kidneys were exposed to 120 min of WI. They were then either reimplanted immediately, hypothermically machine perfused (4 degrees C) for 18 hr with Belzer's solution, or transitioned to 18 hr of warm perfusion (32 degrees C) with an acellular perfusate before implantation. RESULTS: Warm perfused kidneys with 120 min of WI provided life-sustaining function after transplantation, whereas the control kidneys immediately reimplanted or with hypothermic machine perfusion did not. The mean peak serum creatinine in the warm perfused kidneys was 3.7 mg/dl, with the mean peak occurring on day 2 and normalizing on day 9 posttransplant. CONCLUSIONS: These results indicate that 18 hr of ex vivo warm perfusion of kidneys is feasible. Furthermore, recovery of renal function during warm perfusion is demonstrated, resulting in immediate function after transplantation. The use of ex vivo warm perfusion to recover function in severe ischemically damaged kidneys could provide the basis for increasing the number of transplantable kidneys. PMID- 11923689 TI - Cardiovascular morbidity and mortality after orthotopic liver transplantation. AB - BACKGROUND: Hyperlipidemia and hypertension have been reported in liver allograft recipients and contribute to an increased risk of ischemic heart disease (IHD) after orthotopic liver transplantation (OLT). The aims of the study were (1) to determine the prevalence of risk factors for IHD in these patients and (2) to compare the observed incidence of cardiovascular events and related mortality in allograft recipients with a matched population. METHODS: One hundred ten consecutive adults (50 male) who attended for review after OLT (median follow-up 3.9 years; range 0.1-17.9) were assessed for cardiovascular risk factors using current blood pressure, diabetic status, and smoking history and measurements of total cholesterol, high-density lipoprotein cholesterol, and triglyceride concentrations. Cardiovascular events and cardiovascular mortality data were collected from the prospective database of all adult liver allograft recipients and compared to matched data from myocardial infarction registries and Office for National Statistics data, respectively. RESULTS: Raised serum cholesterol (>5.0 mmol/L) was found in 48 (44%) patients (18 male), and systolic hypertension (>140 mmHg) was found in 69 (63%) patients (27 male). The relative risk of ischemic cardiac events was 3.07 (95% [confidence interval] CI, 1.98-4.53) and the relative risk for cardiovascular deaths was 2.56 (95% CI, 1.52-4.05) in allograft recipients compared to an age-matched population without transplants. CONCLUSIONS: Liver allograft recipients have a greater risk of cardiovascular deaths and ischemic events than an age- and sex-matched population. The prevalence of raised cholesterol concentrations in patients after OLT is similar to those in previous reports. Moderate hypertension and hyperlipidemia may be more detrimental in patients after OLT compared to non-transplant patients without these risk factors. PMID- 11923690 TI - Anti-CD20 monoclonal antibody (rituximab) therapy for acute cardiac humoral rejection: a case report. AB - Humoral or antibody-mediated rejection in cardiac transplant recipients is mediated by donor-specific cytotoxic antibodies and is histologically defined by linear deposits of immunoglobulin and complement in the myocardial capillaries. Antibody-mediated rejection often is accompanied by hemodynamic compromise and is associated with reduced long-term graft survival. Standard immunosuppression, designed to target T cell immune function, is largely ineffective against this B cell-driven process. Current treatment options for humoral rejection are limited by a lack of specific anti-B cell therapies. We present the case of a 50-year-old woman with hemodynamically significant humoral rejection resistant to steroids, cyclophos-phamide, and plasmapheresis who responded to the addition of anti-CD20 monoclonal antibody therapy (rituximab). One year posttransplant, the patient is rejection-free, with normal left ventricular systolic function and coronary arteries. PMID- 11923691 TI - Treatment of recurrent allograft dysfunction with intravenous hematin after liver transplantation for erythropoietic protoporphyria. AB - Erythropoietic protoporphyria (EPP) is a rare inherited disorder of the heme biosynthetic pathway in which toxic levels of protoporphyrins often precipitate in the liver, leading to cirrhosis, liver failure, and the need for liver transplantation (OLT). Because the underlying enzyme defect in EPP is bone marrow derived, the risk for recurrent EPP allograft dysfunction is high. Although plasmapheresis may ameliorate acute allograft disease, strategies to maintain disease remission are needed. A 59-year-old man who underwent OLT for hepatic EPP experienced increased bilirubin and aminotransferases on postoperative day 700. Allograft biopsy demonstrated recurrent EPP. He was managed initially with plasmapheresis, hypertransfusion, and infusions of i.v. hematin. After normalization of liver tests, the hematin infusions have been given intermittently, are well tolerated, and associated with normal allograft function for nearly 2 years. This is the first case of the use of hematin given post-OLT to help achieve and maintain remission of allograft EPP disease. PMID- 11923692 TI - Compared to mycophenolate mofetil, rapamycin induces significant changes on growth factors and growth factor receptors in the early days post-kidney transplantation. AB - BACKGROUND: The new immunosuppressive drug Rapamycin (Rapa) is endowed with a mechanism of action that is distinct from that of calcineurin inhibitors. It has been claimed that Rapa does not significantly modulate either the cytokine expression or the transcription of several growth factors in mitogen-activated T cells. Previously, we reported that fine-needle aspiration biopsy (FNAB) sample cultures synthesize a large array of cytokines, and some of them may be powerful predictors of acute rejection in renal transplants. We hypothesized that Rapa may induce significant changes on cytokine production by FNAB sample cultures and on serum cytokine receptors when compared to other immunosuppressive drugs. METHODS: Kidney transplants treated with CsA-Rapa-Pred (Rapa group) were compared with transplants treated with CsA-mycophenolate mofetil-Pred (MMF group). They were studied on day 7 posttransplantation, and they remained rejection free for at least the first 6 months. FNAB samples were cultured and the supernatants were collected at 48 hr of incubation and analyzed by ELISA for interleukin 1 receptor antagonist (IL-1ra), IL-2, IL-6, IL-10, IL-18, monocyte chemotactic protein 1 (MCP-1), soluble tumor necrosis factor I, and transforming growth factor (TGF) beta(1). The soluble receptors for IL-1, IL-2, IL-6, and tumor necrosis factor alpha, together with IL-2 and IL-18 were also measured in serum. RESULTS: Significant differences were observed when comparing Rapa with the MMF group. IL 18 and TGF-beta(1) synthesis were up-regulated, whereas IL-6 and MCP-1 were down regulated in FNAB sample cultures. The Rapa group showed a significant down regulation of each cytokine receptor and of IL-2 in serum. CONCLUSIONS: Rapa was associated with a decreased synthesis of primarily monocyte-derived cytokines and enhanced production of TGF-beta(1), which in an appropriate cytokine milieu may promote allograft tolerance. The down-regulation of cytokine receptors and IL-2 may be associated with a depressed immune response towards the kidney allograft. PMID- 11923694 TI - Obtaining cornea donation consent by telephone. AB - BACKGROUND: Cornea donation process comes up against difficulties in obtaining families' consent. A face-to-face interview is often not possible for logistical reasons. We carried out a prospective study of the effectiveness of telephone contact in obtaining donation consent. METHODS: Consent was obtained by a single, nonmedical, hospital coordinator. If a face-to-face interview was not possible, a telephone interview was conducted using a standardized procedure. RESULTS: Over a 21-month period, 334 families were contacted, either in a face-to-face interview (142, 42.5%) or by telephone (192, 57.5%). Donation consent was obtained in 66.5% of cases, 106 times by telephone and 116 times in face-to-face interview. The acceptance rate was 55.2% by telephone and 81.6% face to face (P<0.001). In total, 47.7% of the cornea recovery consents were obtained after telephone interview. CONCLUSIONS: Telephone interview is an effective method for obtaining consent to cornea donation. Although the acceptance rate using this method is lower than by face-to-face interview, using the telephone should not be overlooked as this enabled procurement of nearly half the corneas in our hospital. PMID- 11923693 TI - Exposure-response relationships for everolimus in de novo kidney transplantation: defining a therapeutic range. AB - BACKGROUND: Exposure, safety, and efficacy data from the two everolimus randomized, double-blind phase 3 trials were evaluated to identify a therapeutic concentration range applicable in de novo kidney transplantation. METHODS: A total of 695 evaluable everolimus-treated patients received either 0.75 or 1.5 mg bid in addition to corticosteroids and cyclosporine (troughs 150-400 ng/ml in month 1 and 100-300 ng/ml thereafter). A total of 3355 everolimus trough levels (Cmin) were obtained in weeks 1 and 2 and months 1, 2, 3, and 6 after transplantation. Each patient's average Cmin was calculated and the values were divided into quintiles: 1.0-3.4, 3.5-4.5, 4.6-5.7, 5.8-7.7, 7.8-15.0 ng/ml (139 patients per quintile). Efficacy was freedom from biopsy-confirmed acute rejection. Safety measures were maximum total cholesterol and triglyceride levels and minimum leukocyte and platelet counts. A sigmoid exposure-response model was used to test the significance of these Cmin-efficacy and Cmin-safety relationships. RESULTS: Freedom from acute rejection was significantly related to Cmin with an incidence of 68% at 1.0-3.4 ng/ml, 81-86% at 3.5-7.7 ng/ml, and 91% at 7.8-15.0 ng/ml (P=0.03). The incidence of hypercholesterolemia, defined as >6.5 mmol/liter, ranged from 76 to 87% over the exposure range without a significant relation to Cmin (P=0.37). The incidence of hypertriglyceridemia, defined as >2.9 mmol/liter, rose from 59 to 77% across the exposure groups (P=0.02). Leukocytopenia, defined as <4x10(9)/liter, occurred in 11-19% of patients across the exposure quintiles showing no relationship to Cmin (P=0.76). The incidence of thrombocytopenia, defined as <100x10(9)/liter, occurred in 2 years=67.3% and 65.8%; <2 years=64.1% and 64.1%). A significant difference in survival was seen in CsA-based immunosuppression (71.2%, 68.1%, 65.4%, and 61%) versus tacrolimus-based immunosuppression (85.8%, 84.7%, 83.3%, and 82.9%) at 1, 3, 5, and 10 years, respectively (P=0.0001). The maximum difference in survival was noted in the first 3 months between CsA and tacrolimus; thus, indicating there may have been other factors (nonimmunological factors) involved in terms of donor and recipient selection and technical issues. The mean annual death rate beyond 2 years posttransplant was 0.47%, with the mean annual death rate for patients who received tacrolimus-based immunosuppression being significantly lower than those who received CsA-based immunosuppression (0.14% vs. 0.8%; P=0.001). The most common etiologies of graft loss were hepatic artery thrombosis (33.4%), acute or chronic rejection (26.6%), and primary nonfunction (16.7%). Of note, retransplantation for graft loss because of acute or chronic rejection occurred only in those patients who received CsA-based immuno-suppression. CONCLUSION: The overall 20-year actuarial survival for pediatric liver transplantation is 64%. Survival has increased by 20% in the last 12 years with tacrolimus-based immunosuppression. Although this improvement may be the result of several factors, retransplantation as a result of acute or chronic rejection has been completely eliminated in patients treated with tacrolimus. PMID- 11923698 TI - Kidney transplantation after liver transplantation from the same donor: four cases of successful steroid withdrawal. AB - BACKGROUND: Administration of corticosteroids to kidney recipients has hampered the complete clinical success of kidney transplantation. Because most organ transplantation in Japan is living-related, we had the experience of performing kidney transplantation (KT) after liver transplantation (LT) from the same donor in four patients and successfully withdrew corticosteroid administration. METHODS: Three pediatric and one adult patient received kidney allografts from 3 to 10 months after LT from the same donor. The immunosuppressive regimen consisted of a corticosteroid and tacrolimus. The steroid was withdrawn after KT in all four patients. After complete withdrawal of the steroid, DNA was extracted from two recipients and examined by polymerase chain reaction to detect microchimerism. A mixed lymphocyte reaction (MLR) and cell-mediated lymphocytotoxicity assay (CML) were performed to test for donor-specific hyporesponsiveness. RESULTS: Steroid withdrawal was successfully accomplished after KT in every patient. No steroid-withdrawal-associated complications were observed. In the three pediatric patients, remarkable catch-up growth was observed after steroid withdrawal. In the two patients tested, donor DNA was not detected by polymerase chain reaction, suggesting the absence of microchimerism. MLR and CML showed that recipient lymphocytes reacted against donor lymphocytes at the same level as against the third-party lymphocytes. CONCLUSION: Steroid withdrawal was successfully achieved in four kidney recipients who had received a liver allograft from the same donor. The MLR and CML findings indicated the absence of donor-specific hyporesponsiveness in vitro. Although the precise mechanism is not yet clear, KT after LT from the same donor should be considered as a method that allows steroids to be withdrawn from the immunosuppressive regimen of KT. PMID- 11923700 TI - Recipient RAS gene variants and renal allograft function. AB - BACKGROUND: Genetic variants of the renin-angiotensin system (RAS) have been implicated in the progression of native kidney diseases. A decreased long-term renal allograft function has also been associated with increased activity of RAS, which may be genetically determined. METHODS: The effect of the angiotensinogen (AGT), angiotensin-converting enzyme (ACE), angiotensin type 1 receptor (AGT1R), and aldosterone synthase (CYP11B2) genotypes on renal function was investigated in 223 first-allograft recipients. Graft function was estimated by yearly determinations of serum creatinine. Genotyping was performed for the M235T-AGT, the I/D-ACE, the A1166C-AGT1R, and the -344T/C-CYP11B2 gene polymorphisms using polymerase chain reaction. RESULTS: The percentage of patients with preserved stable graft function up to 15 years after transplantation was higher when mean blood pressure was <97 mmHg, than when it was >117 mmHg (60 vs. 25% of patients). The CYP11B2 genotype predicted long-term stable graft function with more patients suffering from worsening renal function with the CYP11B2 TT than the CC genotype (P=0.002). There was a weak association between the AGT1R genotype (P=0.037), but not the AGT or ACE genotypes, and a preserved long-term graft function. Cox proportional hazards estimation showed no interactions between the observed effect of CYP11B2 genotype on renal function over time and the number of HLA class I and II matches, other RAS genotypes, graft function, or mean blood pressure at 1 year after transplantation. CONCLUSIONS: The rate of decline in renal allograft function is strongly associated with the CYP11B2 but not AGT, ACE, or AGT1R genotypes. This finding suggests that certain genetic factors related to the RAS are important determinants of long-term renal allograft function. PMID- 11923699 TI - Improved clinical outcomes for liver transplant recipients using cyclosporine monitoring based on 2-hr post-dose levels (C2). AB - BACKGROUND: A prospective, open-label, study was conducted at 29 centers in 9 countries, involving 307 de novo liver transplant patients to compare the clinical usefulness of monitoring 2-hr post-dose cyclosporine (CsA) levels (C2) with conventional trough cyclosporine blood levels (pre-dose) (C0). METHODS: Neoral oral therapy was initiated at 15 mg/kg/day and dose adjusted according to predetermined C2 or C0 target level ranges. The primary efficacy variable was treatment failure at 3 months, where evaluation was based on a composite endpoint of biopsy-proven rejection, treatment for rejection, graft loss, death, or premature withdrawal/discontinuation from the study. RESULTS: Baseline characteristics were similar between groups. Graft loss at 12 weeks (retransplantation or death) occurred in 6.8% C2 and in 7.0% C0 patients. Overall incidence of treated acute rejection was lower for C2 (23.6%) than C0 patients (31.6%) (P=0.144, Cochran-Mantel-Haenszel [CMH] test). In hepatitis C virus (HCV) negative patients, the incidence of rejection in the C2 group was significantly less than in the C0 group (21.2% vs. 33.0%; P<0.05), whereas in HCV-positive patients, the rejection rate was similar in both groups (26.7% for C2 group vs. 27.3% for C0 group: P=0.81). C2 patients (n=16) who reached minimum target CsA levels by day 3 had a notably low incidence of rejection (12.5%), whereas there was no difference in the incidence of rejection in C0 patients, irrespective of time to reach target level. For biopsy-proven acute rejections (21.6% for C2 vs. 30.4% for C0), the incidence of moderate and severe histological diagnosis was significantly lower in the C2 group than in the C0 group (47% vs. 73%; P=0.01). Safety profiles were similar between the two groups, with few patient withdrawals due to adverse events (9.5% for C2; 7.0% for C0). CONCLUSIONS: Using C2 monitoring, the overall incidence of acute cellular rejection was lower compared with the C0 group, and the histological severity of acute rejections was shown to be significantly milder for the C2 group, indicative of good long-term prognosis. These data demonstrate that the use of C2 monitoring is superior to C0 and results in a reduction in the incidence and severity of acute cellular rejection without detrimental effect on the drug safety profile. PMID- 11923701 TI - Complete freedom from rejection after intestinal transplantation using a new tolerogenic protocol combined with low immunosuppression. AB - BACKGROUND: Intestinal transplantation (Itx) remains the most difficult form of transplantation. This is due to the high immunogenicity of the bowel that currently obligates Itx patients to heavy immunosuppression, which causes infection, posttransplant lymphoproliferative disease (PTLD), and drug toxicity. Wider application of Itx depends on the development of tolerogenic strategies to promote engraftment while reducing the need for immunosuppression. We applied a strategy to clinical Itx that combines intraportal donor-specific blood transfusion with a deliberately low immunosuppression protocol (no high-dose steroids; lower tacrolimus level). METHODS: A 55-year-old patient received a combined liver/Itx. Donor-specific whole blood was taken from the donor during procurement and transfused in the recipient portal vein after graft reperfusion. For induction immunosuppression, no intravenous bolus of steroids was given; only two doses of anti-interleukin 2 receptor antibody were administered. The patient received posttransplantation maintenance immunosuppression with lower tacrolimus levels than average (15 ng/ml first month; 5-10 ng/ml thereafter), low-dose azathioprine (1 mg/kg first to third months; 0.5 mg/kg thereafter), and low-dose steroids (Medrol 8 mg twice daily first and second months; 4 mg twice thereafter). The patient was monitored for rejection, graft-versus-host disease, infection, and PTLD. Protocol biopsy specimens were taken from the distal ileum (2 per week). RESULTS: Clinical, endoscopic, and histologic signs of rejection did not develop. Chimerism was identified at day 28. Graft-versus-host disease was absent clinically. Chimerism was self-limiting and disappeared without modifying baseline immunosuppression and without observing a change in graft function. The patient remained free of systemic opportunistic infections, PTLD, and drug toxicity. Total parenteral nutrition was stopped at 7 weeks after transplantation. The patient remains free of total parenteral nutrition and free of rejection at 14 months after transplantation. CONCLUSIONS: We describe an Itx patient who remained rejection free despite receiving significantly lower immunosuppression than average. We hypothesize that intraoperative immunomodulation via intraportal donor-specific blood transfusion in the absence of nonspecific overimmunosuppression promoted Itx acceptance. PMID- 11923702 TI - The clinical use of various blood compartments for cytomegalovirus (CMV) DNA quantitation in transplant recipients with CMV disease. AB - BACKGROUND: The quantitation of cytomegalovirus (CMV) DNA is a cornerstone in the management of CMV disease in transplant recipients. However, a consensus as to what is the optimal blood compartment for the detection and quantitation of CMV DNA in peripheral blood is nonexistent. METHODS: With an automated quantitative assay, we have simultaneously quantified the CMV DNA load in whole blood (WB), plasma (PL), peripheral blood leukocytes (PBL), and peripheral blood mononuclear cells (PBMC) in 319 samples from 17 transplant recipients with 19 episodes of CMV disease that were treated with 2 weeks of intravenous ganciclovir. RESULTS: Higher levels of CMV DNA were observed in WB than PL (PL minus WB mean difference, 0.67 log; 95% confidence interval, -1.02 to -0.32; P=0.0009). This observation was most evident before treatment with intravenous ganciclovir (pretreatment geometric mean CMV DNA was 45,412 copies per ml of WB vs. 14,995 copies per ml of PL). In contrast, the CMV DNA levels between PBL and PBMC were highly comparable throughout the course of CMV disease and its treatment. Intravenous ganciclovir exerted a uniform effect on the four blood compartments with no statistically significant difference in the degree and rate of CMV DNA decline between WB and PL and between PBL and PBMC. CONCLUSIONS: Although our study demonstrates the adequacy of all blood compartments for CMV DNA quantification, the higher sensitivity of WB and its yield of higher CMV DNA render it an optimal sample for monitoring CMV DNA load during CMV disease in immunocompromised patients. PMID- 11923704 TI - Association of CD8+ natural killer T cells in the liver with neonatal tolerance phenomenon. AB - BACKGROUND: Neonatal tolerance is a very interesting phenomenon, because even allogeneic skin grafts are not rejected in these mice at the adult stage. However, the underlying mechanism remains unclear. METHODS: In this study we prepared such tolerant C57BL/6 (B6) mice (H-2b) by the injection of allogeneic lymphocytes of BALB/c origin (H-2d) at the neonatal stage. RESULTS: The total number of liver lymphocytes in these tolerant mice was found to increase when it was examined at the adult stage. Nevertheless, the retention of allogeneic lymphocytes that were injected at the neonatal stage was highest in the spleen. It is speculated that these allogeneic lymphocytes stimulate the hepatic immune system via the portal vein and that such stimulation maintains the tolerance phenomenon. Indeed, these tolerant mice showed elevated levels of IL-2R beta+ CD3int cells (i.e., extrathymic T cells) and NK1.1+ CD3int cells (i.e., NKT cells) in the liver. Even more interestingly, the number and proportion of CD8+ NKT cells, which are usually a minor population in normal mice, increased among NKT cells in the liver of tolerant mice. This became much more prominent when tolerant mice were grafted with allogeneic (H-2d) skin. CONCLUSION: In conjunction with additional data from a cell-transfer experiment and a splenectomy experiment, our results suggest that CD8+ NKT cells in the liver of tolerant mice might be intimately associated with the neonatal tolerance phenomenon. PMID- 11923703 TI - Liver transplantation for Budd-Chiari syndrome. AB - BACKGROUND: Budd-Chiari syndrome (BCS) is a clinical condition characterized by hepatic venous outflow obstruction secondary to an underlying systemic predisposition to thrombosis. METHODS: We reviewed our experience of 19 adult patients who underwent orthotopic liver transplantation for BCS from April 1988 to May 1999 to assess their long-term outcome and specific complications related to this procedure. RESULTS: Of these patients, 13 presented with chronic and 6 with acute liver failure. At presentation predisposing factors included polycythemia rubra vera in five, an undefined myeloproliferative disorder in four, essential thrombocythemia in two, presence of lupus anticoagulant in one, antiphospholipid antibody positivity in one, post-gestational in one, oral contraceptive pill in one, and idiopathic in four. Five patients had undergone previous porto-systemic shunt. Of the 19 patients, 16 are alive at a median follow-up of 89 months (range 1-119) with 2 patients developing disease recurrence at 4 months and 7 years posttransplant, respectively. Four patients have been retransplanted: one for progressive graft dysfunction due to nodular regenerative hyperplasia secondary to azathioprine toxicity, two for hepatic artery thrombosis (one soon after and the other 47 months posttransplant), and one for recurrent BCS. Three patients have died: one from an intra-abdominal bleed secondary to acute hemorrhagic pancreatitis 8 years posttransplant, another from acute myeloid leukemia at 6 years posttransplant, and the third patient from graft failure secondary to severe rejection 1 month posttransplant. CONCLUSION: Liver transplantation for BCS provides good long-term survival with acceptable morbidity. Long-term survival may be prejudiced by progression of the underlying hematological disorders. PMID- 11923705 TI - Modulation of IFN-gamma-induced immunogenicity by phosphatidylethanolamine-linked hyaluronic acid. AB - BACKGROUND: The present study was conducted to examine the possibility of modulating interferon (IFN-gamma)-induced immunogenicity by a novel compound that is composed of a PLA2 inhibitor linked to hyaluronic acid (HYPE). METHODS: HYPE was tested for its effect on IFN-gamma-induced expression of MHC class I, class II, and intercellular adhesion molecule (ICAM-1) in cultured endothelial and renal proximal tubular cells by flow cytometric analysis (FACS) as well as its ability to influence T cell activation in mixed lymphocyte reaction (MLR) or after mitogen stimulation. RESULTS: In FACS, a profound inhibition in MHC class I and ICAM-1 staining was observed in stimulated or unstimulated cells that were incubated with HYPE. This was not due to down-regulation of antigen expression and only occurred when monoclonal antibodies, but not when polyclonal antibodies, were used. HYPE inhibited the induction of MHC class II in both cell types after IFN-gamma stimulation in a dose-dependent manner. Moreover, the induction of class II transactivator (CIITA) was completely inhibited under these conditions, most likely because it blocked the binding of IFN-gamma to the cell membrane. Addition of HYPE to MLR inhibited the proliferation of T cells and the secretion of interleukin (IL)-2, IFN-gamma, and IL-10. This was not observed when HYPE was added together with anti-CD3 or phytohemagglutinin (PHA). CONCLUSION: Our study provides experimental evidence that HYPE has immunosuppressive features. This makes the compound an interesting candidate as an immunosuppressive drug, not only in organ transplantation, but also in diseases where IFN-gamma is overexpressed. PMID- 11923706 TI - A two-step strategy for enlargement of left arterial branch in a living related liver graft with dual arterial supply. AB - The use of small caliber arteries is probably responsible for the higher hepatic artery thrombosis rate initially reported after living related liver transplantation. We described a two-step strategy generating flow-induced enlargement of a small diameter artery in case of left graft dual arterial supply. The smaller arterial branch was ligated during a laparoscopic first-step procedure inducing a 30% enlargement of the remaining branch. The second-step donor hepatectomy was performed 1 week later using a larger artery for successful vascular anastomosis. The flow-induced enlargement of donor hepatic artery may help to reduce hepatic artery thrombosis risk after pediatric living related liver transplantation. PMID- 11923707 TI - Sustained remission of an extensive monoclonal, Epstein-Barr virus-associated diffuse large B cell lymphoma in a kidney-pancreas transplant recipient. AB - BACKGROUND: Epstein-Barr virus associated posttransplant high-grade large cell lymphoma (EBV-PTL) is thought to be incurable solely by cessation of immunosuppression. We report a case of EBV-PTL resolving rapidly after alleviation of immunosuppression. A strong cytotoxic memory T cell (CTL) reaction against two individual recipient EBV strains appears to be involved. METHODS: EBV strain identification in donor lymphocytes, tumor cells of EBV-PTL, and circulating lymphocytes of recipient was performed by latent membrane protein 1 fingerprinting. RESULTS: Stage IIIB EBV-PTL of recipient type was cured through strong autologous CTL reaction consecutive to alleviation of immunosuppression. Latent membrane protein 1 fingerprinting identified three different EBV strains; namely in lymphoma and peripheral lymphocytes of donor and recipient. CONCLUSION: In the setting of solid organ transplantation, rigorous reduction of immunosuppression may lead to cure of high grade EBV-PTL, if the recipient delivers a strong host CTL reaction against the epitopes of EBV latency gene products. Infection with two individual EBV strains earlier in life may increase the number of specific CTL clones and increase the memory capacity. PMID- 11923708 TI - Dendritic cells cross-presenting viral antigens derived from autologous cells as a sensitive tool for visualization of human cytomegalovirus-reactive CD8+ T cells. AB - BACKGROUND: CD8+ T lymphocytes are essential to contain viral infections, such as human cytomegalovirus (HCMV). To visualize these clinically important effector cells, we used dendritic cells (DC), which efficiently present exogenous antigens by MHC class I molecules (cross-presentation). METHODS AND RESULTS: Immature DC were fed with apoptotic debris derived from autologous HCMV-infected macrophages. After maturation, the antigen-loaded DC were used to stimulate lymphocytes from the same donor. We could detect high numbers of interferon-gamma producing CD8+ T cells in HCMV-seropositive but not in HCMV-seronegative healthy individuals and in only one of five HCMV-seropositive immunosuppressed patients. In contrast, autologous DC infected with live HCMV virions stimulated antiviral CD8+ T lymphocytes to a lesser extent. CONCLUSION: We suggest that DC cross-presenting HCMV antigens derived from autologous macrophages could serve as a valuable tool to monitor and possibly enhance the antiviral T-cell response in patients after transplantation. PMID- 11923709 TI - Chlamydia pneumoniae respiratory infection after allogeneic stem cell transplantation. AB - Chlamydia pneumoniae is a common cause of upper and lower respiratory tract infections in immunocompetent patients; however, its role as a respiratory pathogen in immunocompromised hosts has been infrequently recognized. We describe C. pneumoniae lower respiratory tract infection in a 19-year-old male after allogeneic stem cell transplantation. The patient developed fever on day +14, and a subsequent computed tomography scan of the chest revealed a right lateral pleural-based opacity, which was then resected during thoracoscopy. Diagnosis was made by culture and staining of the resected tissue with C. pneumoniae-specific monoclonal antibodies, and azithromycin was administered. To the best of our knowledge, this is the first report of C. pneumoniae respiratory infection after stem cell or marrow transplantation. C. pneumoniae often coexists with other etiologic agents of pneumonia in immunocompromised patients. Considering the infrequency of infections from this organism in this clinical setting, one must still rule out other more likely respiratory pathogens. PMID- 11923710 TI - Tacrolimus-associated severe bilateral corneal ulcer after renal transplantation. PMID- 11923711 TI - Surgical resection of pulmonary metastases after orthotopic liver transplantation for hepatocellular carcinoma. PMID- 11923712 TI - Tacrolimus-induced nephrotoxicity unmasked by induction of the CYP3A4 system with St John's wort. PMID- 11923713 TI - Pitfalls of domino transplant. PMID- 11923715 TI - The role of a clinical doctorate in the advancement of practice. PMID- 11923714 TI - Challenges faced by TVNs in the UK and abroad. PMID- 11923716 TI - Use of vacuum-assisted closure therapy following foot amputation. AB - This case study highlights the use of vacuum-assisted closure (VAC) in a diabetic man following a partial transverse amputation of his foot. In this situation infection-free healing is imperative in order to salvage the limb and prevent further trauma. VAC therapy facilitates rapid granulation of wounds and reduces bacterial colonization rates. This method was adopted as a suitable therapy for treatment of a patient who suffered from a complex wound at high-risk of reinfection. PMID- 11923717 TI - A nurse's aid to clinical selection of pressure-reducing equipment. AB - The selection of appropriate pressure-relieving equipment for individuals in the community is an important process in the prevention of pressure ulcers. The clinical decision is made by community nurses who are accountable for their actions and who therefore have to have clear rationale for the decisions they make. Risk tools are not a completely reliable or valid method of assessing patients. The current national guidelines (National Institute for Clinical Excellence (NICE), 2001) are not specific in relation to the selection or choice of equipment. This article explores how these decisions are made. PMID- 11923718 TI - The tissue viability nurse and effective documentation. AB - The problems associated with inadequate record keeping in tissue viability have been highlighted in a number of related legal proceedings, professional misconduct cases (UKCC 2001a,b), and investigations by the Health Service Commissioner (Ombudsman) (HSC, 2001). Each serves as a reminder of the importance of nursing documentation towards protecting the welfare of patients and clients. A rising tide in clinical negligence (National Audit Office, 2001), and in an increase in the number of nurses removed from the UKCC register (UKCC 2001c), indicate a greater likelihood of nurses (including specialists), being called to provide evidence of standards of care when investigating, predicting and managing clinical risk. This article examines the continued importance of documentary evidence to support such inquiries, clarifying the tissue viability nurse's responsibility in producing accurate and effective records in all aspects of an increasingly expanding role. PMID- 11923719 TI - Debating the relative unimportance of pressure-reducing equipment. AB - This article examines some assumptions underlying the provision of pressure reducing equipment, and argues that failure to 'act first and ask questions later' may be a key source of pressure damage. Indeed, it is argued that prevention of pressure damage can be simplified by identifying three groups of patients: those who have pressure ulcers; those who will develop them in faction is not taken; and those who will not get them. The linkage of risk assessment scores and guidelines is challenge as erroneous and misleading. PMID- 11923720 TI - Pressure ulcer prevention and wound management in Bulgaria. PMID- 11923721 TI - Health inequalities in the firing line. PMID- 11923722 TI - The concept of fatalism and New Labour's role in tackling inequalities. AB - In the past, the focus for Labour administrations' responses to inequalities within the UK has been an anti-poverty strategy. Prime Minister Tony Blair has introduced a fresh perspective on such an analysis by drawing much wider relationship issues, as well as structural ones, to define the problem and therefore come up with the solution. The sound-bite summary of these issues is social exclusion. This expansion of the definition of the problem does have implications for health policy and health workers. The past focus dating from Beveridge (1942) on dealing with disease alone is deemed as a deficient approach. Social exclusion thinking links health into every aspect of the inequalities agenda, in order to deal with the basic problem of 'fatalism'. PMID- 11923723 TI - The practice nurse's role in the management of epilepsy. AB - Epilepsy is a serious and common neurological condition of which the medical, psychological and social implications are far-reaching. This article aims to discuss the practice nurse's role in responding to the needs of clients with epilepsy. This is explored from a primary health care perspective, focusing on nursing interventions and implications for practice. The potential exists for community nursing intervention to improve quality of life for clients through responsive service provision and by challenging the inequality that appears to exist with regard to it. The available research is overtly pharmacological in origin and nursing research in this field is extremely sparse. PMID- 11923724 TI - The pathophysiology of different types of leg ulcers. AB - Leg ulcers are wounds on the leg of 6 weeks or more duration, and may range in size from very small to very large. They are caused primarily by deterioration in the peripheral circulation of the affected individual, although this may arise from a number of conditions. This article discusses the major conditions leading to the formation of a leg ulcer, in order to improve understanding of the rationale for leg ulcer management techniques. PMID- 11923725 TI - The effect of bacteria on leg ulcer healing. AB - It is a common belief that bacterial infection will delay the healing of leg ulcers, or lead to a deterioration in the wound bed or surrounding skin. Despite many years of research however, there is still no agreement on whether this is in fact the case. This article reviews a selection of the available literature regarding the effects of bacteria on leg ulcer healing. Attempts have been made to distinguish differing levels of bacterial presence (e.g. contamination, colonization, infection), but these terms are still loosely applied and consequently ill-defined. Furthermore, the variety of treatment and assessment methods used in the various studies hampers their comparison. The article concludes that future studies should adhere to a uniform methodology in order to provide clear evidence on which to base practice. PMID- 11923726 TI - Preventing unnecessary suffering: an audit of a leg ulcer clinic. AB - Unnecessary patient suffering and increased costs in leg ulcer care were reported in an Effective Health Care bulletin (NHS Centre for Reviews and Dissemination (NHSCRD), 1997). Follow-up assessment of patients with a history of venous ulceration has beneficial effects for patients' quality of life, and for effective and efficient management of resources for community nurses. This article describes the process of implementing evidence-based practice in a leg ulcer clinic in South London within existing resources, which resulted in the successful prevention of leg ulcer recurrence. PMID- 11923727 TI - Head lice: changing the expensive chemotherapy culture. AB - The revulsion prompted by the discovery of head lice on a child's head, and the money spent by the NHS each year on supplying licensed medicines for head lice, is disproportionate to the medical impact that lice have on their hosts. This article discusses the evidence available on the effectiveness of the commonest parasiticidal preparations, and argues the case for a more rational, simpler and cost-effective response to this common problem. PMID- 11923728 TI - Implementing the NHS plan. PMID- 11923729 TI - Patients must be treated according to clinical need. PMID- 11923730 TI - Nurses must enhance care in the wake of Bristol. PMID- 11923731 TI - Pain management 2: the use of pyschological approaches to pain. AB - This article is the second of a two-part series looking at the psychological and social factors which affect pain perception. The first article (Vol 10(14): 903 11) examined neurophysiological and psychological factors of pain perception. This article considers the importance of the nurse-patient interaction in the management of patients' pain. It outlines the adoption of several psychological approaches which could be utilized by nurses when dealing with patients in pain. The need to encourage a feeling of control over pain is discussed in relation to the use of patient-controlled analgesia and the development of positive coping skills. The value of good communication skills is emphasized as being essential in encouraging patients in pain to utilize these psychological approaches. PMID- 11923732 TI - Caring for men with breast cancer: causes, symptoms and treatment. AB - Male breast cancer is relatively rare when considering more common cancers that affect men such as lung cancer. Approximately 200 men are diagnosed each year in the UK with breast cancer. This article outlines some of the causes of male breast cancer: physiological changes, oestrogen-producing tumours, and certain drugs. Risk factors such as a particular hereditary gene, age, ethnic factors, and geographical variations are discussed. Symptoms that men may present with are described and nurses are asked to encourage men to seek early advice if they have any concerns regarding their breasts. Treatment options are outlined; these treatments are very similar to those offered to women with breast cancer. However, it must be noted that because male breast cancer is uncommon it is difficult to accumulate extensive data concerning the condition. Finally, the important role the nurse plays in supporting men with breast cancer is discussed. Nurses often act as advocates for women with breast cancer and this article suggests that they are in an ideal position to do the same for men with breast cancer. PMID- 11923733 TI - The NSF for mental health from a transcultural perspective. AB - Since 1997, the new Labour Government has taken steps to revitalize the NHS in an attempt to ensure a safe, sound and supportive service for all of its customers. The drive towards an efficient and productive system of health care has been echoed through policy directives with operational strategies, as evident in various Department of Health (DoH) publications, including the NHS Plan (DoH, 2000). The National Service Framework (NSF) for Mental Health (DoH, 1999a) sets out a broad vision for the future, a vision where principles of care, quality and standard of services for the mentally ill are optimized. However, this vision is short of ideas which implicitly address issues directly related to the mental health needs of minority ethnic groups. As an area which is already exposed to severe criticism, this article seeks to adopt a 'transcultural' perspective on the NSF, exploring issues deemed necessary for the success of the NSF in meeting the needs of ethnic minorities. An initial examination of current problems sets the scene for future direction where principles of partnership in care, antioppressive practice, transcultural nursing, and education and training are seen as imperatives which need exploration. The author argues that it is imperative that these principles are fully examined and implemented into practice, to ensure a service based on quality standards and equity principles. For nurses it means the development and learning of new working principles and skills. PMID- 11923734 TI - Rapid urinalysis assays for the diagnosis of urinary tract infection. AB - Urinary tract infection (UTI) in premenopausal women is a frequent complaint in general practice. UTI is usually diagnosed on the basis of clinical symptoms and the use of one or more laboratory tests, the most common being rapid urinalysis reagent assays (urine dip) or midstream urine culture. In order to correlate the leucocyte esterase results of a rapid urinalysis assay with direct urine microscopy for pyuria, undiluted non-centrifuged urine samples from 206 volunteer female healthcare professionals were subjected to analysis using direct urine microscopy using a counting chamber and a rapid urinalysis assay. Of the 206 specimens, 74 were positive for leucocyte esterase using the rapid urinalysis assay, and 39 specimens demonstrated significant pyuria (greater than or equal to 10 leucocytes/microl) on direct microscopy. When the leucocyte esterase results were correlated with the direct urine microscopy results, an assay reading of 15 leucocytes/microl ('trace' on the visual scale) had a sensitivity of 91%, specificity of 79%, positive predictive value of 53% and a negative predictive value of 97%. An assay reading of 25 leucocytes/microl ('+' on the visual scale) or greater had a sensitivity of 63%, specificity of 95%, positive predictive value of 75% and a negative predictive value of 91%. In premenopausal, non pregnant females, a rapid urinalysis assay result of 25 leucocytes/microl or greater will predict significant pyuria on urine microscopy with reasonable confidence, thereby reducing the need for more costly urine cultures. PMID- 11923735 TI - Legal aspects of consent 11: compulsory Caesarean sections. AB - Case Scenario: Miss MB required a Caesarean section in order to save her fetus. However, while she gave consent to the operation, she suffered from a needle phobia which caused her to panic and refuse the preliminary anaesthetic. The trust applied for a declaration that the Caesarean section could take place on the grounds that the needle phobia rendered her mentally incapacitated and therefore the operation should proceed in her best interests (Re MB (an adult: medical treatment), [1997]). PMID- 11923736 TI - Dansac Invent convex in the management of flush ileostomy. AB - This article examines the use of the Dansac Invent convex drainable stoma appliance. A review of the literature highlights the indications for use of convexity and discusses the potential contraindications. A short case study describes the use of convex appliances in a patient with a flush, high-output ileostomy. PMID- 11923737 TI - Doctors' education needs to be more holistic. PMID- 11923738 TI - What role for the arts in palliative care? PMID- 11923739 TI - Pressure ulcer prevention in palliative care 1: results of a UK survey. AB - Pressure ulcer prevention is an important aspect of palliative care nursing. This article reports part 1 of the results of a postal questionnaire survey of all 206 palliative care inpatient units listed in the St Christopher's' information service hospice directory. The key areas surveyed were current pressure ulcer prevention policies in use in UK palliative care units and current practice in relation to pressure ulcer risk assessment. Issues arising from these results are also discussed: the key elements of a pressure ulcer policy and the role and timing of pressure ulcer risk assessment. PMID- 11923740 TI - The measurement, causes and effective management of cancer-related fatigue. AB - A review of research into cancer-related fatigue undertaken since 1995 is presented. The manner in which such fatigue varies with cancer diagnosis, stage of disease and anti-cancer treatment is discussed, and the causes of cancer related fatigue are categorized according to whether they are cancer-specific, common to other chronic illnesses or common to the general population. Interventions to alleviate fatigue are discussed in terms of whether they are pharmacological or non-pharmacological in nature. It is concluded that cancer related fatigue is a common problem with a major impact on quality of life. It shares a common aetiology with other forms of fatigue. Graded aerobic exercise has been shown in randomized controlled trials to be an effective intervention in specific patient groups. Less direct evidence supports the use of psychological interventions, but there is very little evidence to support the use of pharmacological treatment, with the possible exception of erythropoietin therapy for anaemic patients undergoing chemotherapy. PMID- 11923741 TI - Music therapy in palliative care: current perspectives. AB - As the music therapy profession has developed internationally over the last 25 years, so has its role in palliative care. Music is a highly versatile and dynamic therapeutic modality, lending itself to a variety of music therapy techniques used to benefit both those living with life-threatening illnesses and their family members and caregivers. This article will give a broad overview of the historical roots of music therapy and introduce the techniques that are employed in current practice. By combining a review of mainstream music therapy practice involving musical improvisation, song-writing and receptive/recreational techniques with case material from my own experience, this article aims to highlight the potential music therapy holds as an effective holistic practice for palliative care, whatever the care setting. PMID- 11923743 TI - A philosophical discussion of end-of-life decision-making methods for incompetent patients. AB - This article provides an analysis of decision-making methods that can be used with incompetent patients (patients who are unable to make decisions for themselves). It is intended as a general discussion of the relevant issues rather than an examination of legal or policy concerns. The advance directive principle (whereby a person leaves instructions for what to do if they become unable to make their own decisions) is discussed with reference to practical and philosophical issues. Substituted judgement (where surrogate decision makers make decisions that the patient who is incompetent would make if competent) is then discussed followed by the principle of best interests, which requires consideration beyond purely medical interests. The need to make patient-centred quality-of-life judgements as a component of best interests decisions is argued. Conclusions reached include the notion that the advance directive principle and the substituted judgement principle facilitate the concept of self-determination. The best interests principle helps to ensure that patients ending up in this vulnerable position are protected. PMID- 11923742 TI - Perception of empowerment by family caregivers of patients with a terminal illness in Hong Kong. AB - The purpose of this study is to provide an in-depth description of the nurse facilitated empowering experiences of family caregivers of terminally ill patients in Hong Kong. Data were collected through in-depth interviews with 24 family caregivers of patients with terminal cancer. Grounded theory strategies served as the basis for data collection and analysis. Data were analysed in terms of the strategies, consequences, and interactions involved in the empowering experience. Themes included establishing an engaged relationship; providing information, knowledge and skills; affirming self-worth; and giving reassurance that the patient is under good care. The consequences of empowerment included a trusting relationship, having confidence to carry out caregiving tasks, acceptance of patient's death, sustained self-worth and feeling peace at heart. The focus of empowering experiences was related to the quality of life of the dying patients. Family caregivers require information, education, encouragement and support, and nurses are in an ideal position to meet this need. Fundamental in the empowering process is the formation of a trusting relationship and as a result the caregivers feeling they are not being abandoned or left alone in the caring process. The results of this study can help nurse interact more effectively with family caregivers, giving them direction for effective nursing practice when caring for patients with a terminal illness. PMID- 11923744 TI - Why not natural death? PMID- 11923745 TI - Food refusal and dysphagia in older people with dementia: ethical and practical issues. AB - Food refusal poses difficulties for nurses and care staff and can place older patients with dementia at risk of undernutrition. The dangers of and reasons for food refusal in these patients are explored and the fundamental ethical and legal issues raised in caring for these patients are examined. Practical guidance and solutions are offered through dietetics and speech and language therapy with the aim of helping nurses and care staff, as well as professional carers in palliative care, and explore ways of critically examining and coping with these dilemmas. PMID- 11923746 TI - Celebrating 150 years of care at the Royal Marsden Hospital. PMID- 11923747 TI - The nature of terminal malignant bowel obstruction and its impact on patients with advanced cancer. AB - Patients with bowel obstruction due to advanced cancer often have a poor prognosis and suffer distressing symptoms that can be difficult to manage. Research to date has focused on medical interventions and the measurement and control of symptoms. Little attention has been given to patients' perceptions of their quality of life or the impact of their illness. This study aimed to explore patients' lived experience of bowel obstruction. Semi-structured interviews were conducted with ten patients with bowel obstruction due to gynaecological or gastric cancer. Data analysis was based on an adaptation of Giorgi (1975) and Parse et al's (1985a,b) phenomenological method. Findings suggest that the 'meaning' of being unable to eat is more significant for patients than the nutritional or biological loss of food. For some patients, bowel obstruction marks a process of transition from life to death which is characterized by a sense of social disengagement and disrupted identity. Interviews highlighted aspects of the nurse-patient relationship that were therapeutic in their own right. PMID- 11923748 TI - Presenting a case for involving children with a terminal illness in clinical trials. AB - Today's approaches to the treatment of childhood malignancies are the result of past clinical trials. In order to improve survival it is essential we continue to seek benefit from clinical trials. However, entry of terminally ill children into a phase I or phase II clinical trial, involving a novel, potentially therapeutic, agent is highly contentious. This article will argue in favour of a child's active and full participation in the decision to enter into a phase I or phase II clinical trial. A formal study of the role of children in clinical trials, especially phase I and phase II trials, is urgently required if standards of best practice are to be laid down and subsequently measured. PMID- 11923749 TI - A model of palliative care for the adolescent with cancer. AB - Adolescents are a distinct group in paediatric and adult cancer and palliative care specialities. The process that is experienced by the patient and his/her family of the transition from health to living with a life-threatening illness and from a life-threatening to a life-limited illness raises specific issues for service provision. The adolescent population presents with a wide variety of physical and emotional maturity that highlights the need for health professionals to be equipped with skills in adolescent care, cancer care and specialist palliative care. Due to the small number of patients who will require such a service in any one health district in the UK this is not usually realistic. This article discusses a model of care that promotes collaborative professional practice in a cancer centre between the paediatric department and adult specialist palliative care team that in turn extends a philosophy of care into cancer units and the community setting. PMID- 11923750 TI - The ethics and practicalities of consent in palliative care research: an overview. AB - In order to ensure that further developments and improvements are made in palliative care, research is essential. Palliative care is no different from other specialities in that it needs a scientific foundation on which to base its practice. Research in palliative care is particularly difficult, however, because of the population under study. Research in palliative care presents a 'minefield' of ethical issues. One of the major issues is how to obtain informed consent from patients. This article discusses the practical and ethical issues surrounding consent for quantitative research in palliative care, and offers some guidance to health professionals considering the issue with patients. PMID- 11923751 TI - Guidelines in respect of advance directives: the position in England. AB - The fundamental aim of the advance directive is to provide a means for the patient to continue to exercise autonomy and shape the end of his/her life. The principle is not new. Patients who are aware of approaching death have for many years discussed with their health-care team how they wish to be treated. The advance directive registers these views in a more formal way. However, inconsistencies persist in the understanding of the legal issues around advance directives in the UK, and suggest that national guidelines would encourage National Health Service trusts to develop local policies to improve understanding and compliance with the law. This article draws on English law to address misunderstandings and proposes which information advance directives should include. PMID- 11923752 TI - Supportive and palliative care networks: a new model for integrated care. AB - Cancer is one of the biggest killers in the UK, and recent Government health policy has responded to this with a number of specific developments. Foremost among these is 'The NHS Cancer Plan' (Department of Health, 2000), which established cancer networks as a new model of integrated cancer care service provision. In response, palliative care providers have initiated the development of networks for supportive services of patients alongside the cancer networks to improve coordination of care. This article will briefly discuss the parallel development of cancer and supportive care networks and examine what form the latter will eventually take. PMID- 11923753 TI - Enhancing creativity to improve palliative care: the role of an experiental self care workshop. AB - Creativity plays an important part in all aspects of life. Although the nature of creativity is hypothesized in a variety of ways, there is no common agreement as to how or why it manifests. In the USA and UK, creative ability is often restricted by education, and nursing education programmes appear to be no exception. This article discusses the development of a weekend workshop programme for health professionals working with older people and in palliative care, using Alligood's (1991) hypotheses, that there are positive correlations between creativity, self-actualization and empathy. Actualization and creativity are closely related to self-care and there is evidence that 'selfishness' is an essential ingredient in the actualizing process. There is also evidence that an empathic relationship between health workers and their patients has a significant positive impact on interpersonal care processes. If the creative potential of nurses and carers can be enhanced through self-care, their ability to empathize may be increased. The focus of the workshops is on creative methods of self-care, using experiential exercises in relaxation, music, touch, massage and silence. Evaluation of the workshops revealed that the potential of using this method to develop care practices is worthy of investigation and development. PMID- 11923755 TI - A randomized trial comparing injection therapy with hemoclip and with injection combined with hemoclip for bleeding ulcers. AB - BACKGROUND: A randomized comparative study was conducted of injection therapy with epinephrine-polidocanol (1%) versus hemoclip application, versus injection combined with hemoclip for bleeding peptic ulcers. METHODS: One hundred five patients were randomized and 101 could be evaluated (46 had active spurting or oozing of blood; 55 a visible vessel). Patients were randomized to 1 of the 3 treatment modalities during endoscopy performed within 12 hours of admission. Endoscopy was repeated after 1 day or at recurrence of bleeding and before discharge. In case of recurrent bleeding, patients were retreated with the same modality. RESULTS: Initial failure or the rate of early recurrence of bleeding was highest (but not statistically significant) in the hemoclip group (13/35; 37%), versus the injection (5/34; 15%) and combination (8/32; 25%) groups. Overall failure was significantly (p = 0.01) different among the 3 groups with the highest rate in the hemoclip group (12/35; 34%), versus the injection (2/34; 6%) and combination therapy (8/32; 25%) groups. The use of hemoclips alone appeared to fail because of difficulty with hemoclip placement and incomplete vessel compression. Complications included 1 perforation in the injection group and possibly 1 case of septic arthritis in the combination therapy group. CONCLUSION: In this study, endoscopic treatment of bleeding peptic ulcers with the hemoclip was inferior overall to injection therapy. PMID- 11923754 TI - Empirical treatment based on "typical" reflux symptoms is inappropriate in a population with a high prevalence of Helicobacter pylori infection. AB - BACKGROUND: Empirical therapy or early endoscopy have been recommended as acceptable management options for GERD. The objective of this study was to determine whether diagnosis and empirical treatment based on reflux symptoms alone are appropriate as initial management for patients with gastroesophageal reflux. METHOD: Consecutive patients presenting with weekly reflux symptoms were evaluated with a structured questionnaire followed by endoscopy. Patients with dyspepsia as the predominant symptom, "alarm" symptoms (weight loss, dysphagia, or bleeding), history of peptic ulcer or gastric surgery, or recent nonsteroidal anti-inflammatory drugs intake were excluded. RESULTS: Four hundred sixty patients were studied: 82 (18%) were found to have peptic ulcer disease and 78 (95%) were infected with Helicobacter pylori. Concomitant erosive esophagitis was found in 26 (32%) of these patients with peptic ulcer disease. In the remaining 378 patients, 218 (58%) had erosive esophagitis and 1 had esophageal cancer. Among the 159 patients with no endoscopic lesion, 148 (93%) had relief of symptoms when treated with a proton pump inhibitor. Multivariate analysis showed that male gender (OR: 1.8, p = 0.03), age greater than 60 years (OR: 2.2, p = 0.01) and H pylori infection (OR: 3.6, p = 0.008) were significantly associated with a diagnosis of peptic ulcer disease. Coexisting dyspeptic symptom was not a predictor (p = 0.13) for peptic ulcer disease. CONCLUSIONS: In populations with a high prevalence of H pylori infection, a significant proportion of patients with GERD have concomitant peptic ulcer disease. Empirical treatment based on "typical" GERD symptoms alone may not be appropriate. PMID- 11923756 TI - Octreotide versus hydrocortisone versus placebo in the prevention of post-ERCP pancreatitis: a multicenter randomized controlled trial. AB - BACKGROUND: Octreotide is a potent inhibitor of pancreatic secretion, and corticosteroids suppress humoral and cellular activity. Both agents may reduce the frequency of post-ERCP pancreatitis. The aim of this study was to determine the effectiveness of octreotide and hydrocortisone in preventing post-ERCP pancreatitis. METHODS: Three hundred fifty-four patients were entered in to a multicenter randomized controlled trial of 100 microg subcutaneous octreotide (Group 1) versus 100 mg intravenous hydrocortisone (Group 2) versus normal saline solution as placebo (Group 3). All medications were administered approximately 30 minutes before the procedure. Patients were assessed clinically and serum amylase was also measured before the procedure and 3, 12, and 24 hours after the procedure. RESULTS: Three hundred forty patients were included in the analysis. Pancreatitis was observed in 11 of 112 patients (9.8%) in Group 1, 8 of 113 (7.1%) patients in Group 2, and in 15 of 115 (13.0%) patients in Group 3 (p = 0.32). The mean length of hospitalization in days was similar in all 3 groups: mean (SD) for Groups 1, 2, and 3 were, respectively, 3.6 (1.6) versus 2.9 (0.6) versus 4.3 (1.8) (p = 0.13). Multivariate logistic regression analysis showed that number of pancreatic injections, suspicion of sphincter dysfunction, therapeutic procedure, and age were risk factors for pancreatitis. CONCLUSIONS: The results of this trial indicate that octreotide and hydrocortisone do not prevent ERCP-induced pancreatitis. PMID- 11923757 TI - Complications of endoscopic sphincterotomy: can heparin prevent acute pancreatitis after ERCP? AB - BACKGROUND: An exploratory analysis of a prospective study of risk factors for acute pancreatitis after ERCP combined with endoscopic sphincterotomy showed that the frequency of acute pancreatitis was lower in patients who received heparin compared with patients not treated with heparin. The study was continued to further analyze the effect of heparin on the frequency of acute pancreatitis. METHODS: Potential risk factors for acute pancreatitis and outcomes were evaluated prospectively for all ERCP procedures with endoscopic sphincterotomy performed between September 1994 and December 1998. The results were analyzed by univariate and multivariate methods to determine risk factors for complications. Heparin was administered to 32.9% of the patients (heparin group [HEP group], n = 268) for various clinical reasons (low-molecular-weight heparin, n = 208, unfractionated heparin n = 60). A group of 547 patients who did not receive heparin served as control patients (CON group). RESULTS: Eight hundred fifteen patients underwent ERCP with endoscopic sphincterotomy; acute pancreatitis occurred in 6.4% (n = 52). The frequency of acute pancreatitis was significantly lower in the HEP group versus the CON group in the final multivariate model, which included significant risk factors for acute pancreatitis (HEP group: 3.4%, 9/268 vs. CON group: 7.9%, 43/547; p = 0.005). HEP did not increase the risk of hemorrhage (HEP group: 1.1%, 3/268, 2 severe, none fatal vs. CON group: 2.0%, 11/547, 3 severe, 2 fatal). HEP (p = 0.005; OR 0.3: 95% CI [0.16, 0.73]) and the number of risk factors present (p = 0.0001; OR 2.5: 95% CI [1.80, 3.50]) influenced the frequency of acute pancreatitis independently. CONCLUSIONS: Heparin was significantly associated with an extremely low frequency of post-ERCP pancreatitis without increasing the risk of hemorrhage after endoscopic sphincterotomy. Because this effect could not be attributed to other known or suspected confounders, our conclusion was that heparin administration before ERCP reduces the risk of pancreatitis. PMID- 11923758 TI - Accuracy of a narrow-diameter battery-powered endoscope in sedated and unsedated patients. AB - BACKGROUND: Esophagoscopy with a portable battery-powered endoscope could provide a safe, inexpensive, and minimally invasive way to screen for Barrett's esophagus or esophageal varices. The use of such an instrument in an unsedated fashion has not been previously evaluated. METHODS: Patients referred for an EGD were recruited to undergo an additional examination with the battery-powered endoscope before EGD. In phase 1, (n = 42) patients received conscious sedation before the battery-powered endoscopic examination. In phase 2, (n = 56) patients were not sedated and were given the option of a peroral (n = 43) or transnasal (n = 13) endoscopy. Examiners were blinded to patient history and procedure indications. Esophageal findings, ease of intubation, optical quality, and patient comfort for the battery-powered endoscope and standard EGD were recorded by the endoscopist. RESULTS: Ninety-eight patients (60 men, 38 women, mean age 53 years) were recruited. The sensitivity for detecting Barrett's esophagus, esophageal tumors, and esophageal varices was 54.5%, 66.7%, and 80%, respectively. Ease of intubation and patient comfort as perceived by the endoscopist were not significantly different between the battery-powered endoscope and EGD. Optical quality was ranked as less than 4 (on a 5-point scale with 5 = standard EGD and 1 = poor) in 42% of battery-powered endoscopic examinations. There were no complications. CONCLUSION: The accuracy of esophageal examination with a 3.1-mm endoscope is substantially inferior to standard EGD. Thus, the battery-powered endoscope would not be useful for screening patients for Barrett's esophagus or varices unless improvements in optical quality and visualization are made. PMID- 11923759 TI - Endotherapy of early onset idiopathic chronic pancreatitis: results with long term follow-up. AB - BACKGROUND: Idiopathic chronic pancreatitis that presents at age 35 years or younger has been classified as early onset type and is often characterized by chronic severe pain. Endotherapy, with drainage of the main pancreatic duct, can lead to control of pain if ductal hypertension is an important cause. Long-term results of endotherapy in patients with early onset idiopathic chronic pancreatitis are reported herein. METHODS: This retrospective study consists of 11 patients (6 men, 5 women; mean age 24.2 years, range 16-34 years) treated endoscopically in a 6.5-year period. The indication for treatment was pain in all patients and all had a dilated main pancreatic duct on pancreatography. The objectives of endoscopic treatment were to obtain good drainage of the pancreatic duct and complete clearance of ductal stones. RESULTS: Treatment was successful in all patients with no procedure-related mortality and with mild complications. Seven patients remained free of pain relapses after a mean follow-up of 78.3 months (37-116 months). Seven relapses of pain were recorded in the remaining 4 patients. Endoscopic retreatment was successful in all cases. The difference between the number of hospitalizations during the year before treatment (mean 2.2, range 1-9) and the year after (mean 0.3, range 0-2) was statistically significant (p < 0.01). Statistical significance was maintained at 3 and 6 years' follow-up. CONCLUSIONS: Endoscopic treatment could be regarded as the initial management of choice for patients with early onset idiopathic chronic pancreatitis. PMID- 11923761 TI - A prospective evaluation of outcome in patients referred for PEG placement. AB - BACKGROUND: PEG feeding is not recommended for short-term use because the 30-day mortality after PEG placement is substantial. The primary aim of this study was to prospectively identify factors predictive of survival in patients referred for PEG placement. METHODS: All patients for whom gastroenterology consultation was sought for feeding PEG placement were prospectively studied. Demographic data, Charlson comorbidity index, and functional status were recorded at entry. After PEG placement, patients were followed for up to 12 months. RESULTS: Of the 67 patients for whom consultation was requested, 58 were eligible for the study and 50 underwent PEG placement. The 7-day and 30-day mortality rates in the PEG placement group were 4% and 20%, respectively. In multivariate analysis, only the Charlson index > or =4 was associated with decreased survival time (relative hazard = 2.9: 95% CI [1.20, 7.21], p = 0.019). Median survival in patients with Charlson comorbidity index > or =4 was significantly shorter than that in patients with Charlson index < 4 (p = 0.013). CONCLUSIONS: A Charlson comorbidity index > or =4 was significantly associated with shorter patient survival after initial consultation. Careful consideration of predictive factors of survival may improve patient selection for feeding PEG placement. PMID- 11923760 TI - ERCP features in 27 patients with autoimmune pancreatitis. AB - BACKGROUND: Autoimmune pancreatitis has been proposed as a new clinical entity. However, the ERCP features of this entity have not been well described. METHODS: Clinical and radiographic features in 27 patients with a diagnosis of autoimmune pancreatitis were evaluated. RESULTS: All 27 patients were at first suspected to have pancreatic cancer. The patients were predominantly elderly men and presented with jaundice or mild symptoms and pancreatic enlargement, but no attack of acute pancreatitis. A cholestatic biochemical profile and serum IgG elevation were usually present. A diffusely or segmentally irregular and narrow main pancreatic duct and a positive response to corticosteroid therapy were characteristic. During observation without treatment, serial pancreatography in 2 patients demonstrated progression of diffuse, irregular narrowing of the main pancreatic duct over periods, respectively, of 2 and 6 months. Both the irregular narrowing of the main pancreatic duct and distal bile duct strictures improved in various degrees in response to corticosteroid therapy. CONCLUSIONS: Segmental or diffuse irregular narrowing of the main pancreatic duct are ERCP features of autoimmune pancreatitis. PMID- 11923762 TI - EUS in the diagnosis of early chronic pancreatitis: a prospective follow-up study. AB - BACKGROUND: The aim of this prospective, follow-up study was to investigate the value of EUS in the diagnosis of alcohol-induced chronic pancreatitis. METHODS: One hundred thirty patients with known (n = 51) or clinically suspected (n = 79) chronic pancreatitis were included. Patients with a history of chronic use of alcohol and recurrent abdominal pain underwent endoscopic retrograde pancreatography and EUS. The 38 patients with normal endoscopic retrograde pancreatography but signs of chronic pancreatitis on EUS were included in a follow-up program. RESULTS: All patients with chronic pancreatitis confirmed by retrograde pancreatography (n = 92; 70.8%) had ductal or parenchymal changes detectable with EUS. Among 38 patients (29.2%) with normal retrograde pancreatography, 32 (84.2%) presented with morphologic features consistent with chronic pancreatitis by EUS. During follow-up (median 18 months, range 6-25 months) chronic pancreatitis was confirmed by repeat endoscopic retrograde pancreatography in 22 of these 32 patients (68.8%). On the basis of these follow up data, the sensitivities of EUS and endoscopic retrograde pancreatography at the time of the first examination were, respectively, 100% and 80.7% (p < 0.001). CONCLUSION: EUS detects chronic pancreatitis in all cases if endoscopic retrograde pancreatography was suggestive for chronic pancreatitis. However, EUS is more sensitive than endoscopic retrograde pancreatography in the detection of early morphologic changes of chronic pancreatitis in patients with abdominal pain and a history of chronic and continued ingestion of alcohol. PMID- 11923763 TI - Hemodynamic assessment of the left gastric vein in patients with esophageal varices with color Doppler EUS: factors affecting development of esophageal varices. AB - BACKGROUND: An understanding of the development of esophageal varices is important in the evaluation of risk of variceal hemorrhage. To clarify factors affecting the development of esophageal varices, the morphology and hemodynamics of the left gastric vein were analyzed with color Doppler EUS. METHODS: Sixty seven patients with esophageal varices underwent color Doppler EUS. Seventeen had small varices (F1), 32 had medium varices (F2), and 18 had large varices (F3). RESULTS: Hepatofugal blood flow velocity in the left gastric vein trunk increased as the size of the varices increased (p < 0.0001), whereas the diameter did not increase. The left gastric vein bifurcates into anterior and posterior branches. As the size of the varices enlarged, the branch pattern was more likely to be anterior branch dominant (p = 0.041). There was no significant difference between the 3 size groups of esophageal varices with respect to the size of the paraesophageal collaterals. The detection rate and diameter of the perforating vein increased as the size of the varices increased (p = 0.032 and 0.012, respectively). CONCLUSION: Blood flow velocity in the left gastric vein trunk, branches, and perforating veins may regulate blood flow supplying the esophageal varices and contribute to their development. These findings are important to understanding the pathogenesis of esophageal varices. PMID- 11923764 TI - Duration of antibiotic therapy for cholangitis after successful endoscopic drainage of the biliary tract. AB - BACKGROUND: Drainage of the obstructed biliary tree is the mainstay of therapy for patients with acute cholangitis; antibiotic therapy is complementary. It is unknown whether it is necessary to continue therapy with antibiotics once biliary drainage is achieved and signs of systemic inflammation have subsided. METHODS: Patients who presented with acute cholangitis and were successfully treated at ERCP were studied retrospectively. Patients were followed for 6 months after ERCP. RESULTS: Eighty patients fulfilled study criteria. In 46% of patients blood cultures grew microorganisms. All patients recovered from the episode under study. Antibiotic therapy after ERCP was given for a median duration of 3 days (range: 0-42). Forty-one patients received antibiotic therapy for 3 days or less, 19 for 4 or 5 days, and 20 patients longer than 5 days. The 3 groups were well matched. In none of the patients did the index episode of cholangitis result in a secondary complication not present at the time of ERCP. The percentage of patients with recurrent cholangitis (24%) was not statistically different for the 3 groups (p = 0.80). CONCLUSIONS: Short-duration antibiotic therapy (3 days) appears sufficient when adequate drainage is achieved and fever is abating. PMID- 11923765 TI - Do patients with recurrent choledocholithiasis after endoscopic sphincterotomy benefit from regular follow-up? AB - BACKGROUND: As many as 24% of patients who undergo endoscopic sphincterotomy for the removal of bile duct stones have recurrent biliary complications develop for which there is no effective method of prevention. The aim of this study was to determine whether patients who undergo routine clinical follow-up after endoscopic sphincterotomy for bile duct stones have a different outcome than those who do not. METHODS: All patients who had endoscopic sphincterotomy for bile duct stones were scheduled for follow-up visits, liver function tests, and transabdominal US every 3 to 6 months. ERCP was performed whenever symptoms recurred, or abnormal liver function or US was noted. The patients themselves decided whether to adhere to the follow-up schedule; this was not a randomized trial. RESULTS: Seven hundred sixty-seven patients underwent endoscopic sphincterotomy for bile duct stones from October 1990 to July 1999. Seventy-seven (10%) were found to have recurrent bile duct stones. Three patients who had undergone Billroth II gastrectomy were excluded. Among the remaining 74 patients (52 men, 22 women; mean age 65 years), 42 (57%) had a juxtapapillary diverticulum and 21 (28%) an intact gallbladder. The mean time to recurrence of bile duct stones was 19.7 months (range 5-72 months). Sixty-four patients (87%) had recurrent bile duct stones within 3 years. Fifty-one (69%) were followed regularly (Group A) and 23 (31%) were not (Group B). At the time of stone recurrence, 20 patients in Group A (39%) and 1 in Group B (4%) were asymptomatic (p = 0.002). Liver function tests were normal in 17 patients (13 Group A, 4 Group B). Endoscopic treatment for recurrent bile duct stones was successful in 46 patients (90%) in Group A and 16 (70%) in Group B (p = 0.04); surgical treatment was successful in all 5 patients in Group A and 4 of the 7 patients in Group B. Two patients in Group B were treated by nasobiliary drainage and biliary endoprosthesis insertion caused by extremely large stones and poor condition; both died (acute pancreatitis and sepsis). CONCLUSION: Regular follow-up after endoscopic sphincterotomy detects recurrent bile duct stones early and thus avoids complications of bile duct stones. PMID- 11923766 TI - Endoscopic management and long-term follow-up of Dieulafoy's lesions in the upper GI tract. AB - BACKGROUND: The aim of this study was to retrospectively evaluate the short-and long-term effectiveness of different methods of endoscopic treatment for bleeding Dieulafoy's lesions. METHODS: Patients were allocated into 2 groups according to the hemostatic method applied: (1) injection group (epinephrine and/or ethanolamine oleate), and (2) thermal coagulation group (heat probe), either alone or combined with epinephrine injection. The combination of epinephrine and ethanolamine oleate was used in 5 patients, epinephrine alone in 3, ethanolamine oleate alone in one, heat probe and epinephrine in 8, and heat probe alone in 1 patient. RESULTS: Dieulafoy's lesions were found in 18 (1%) of 1750 patients with acute nonvariceal upper GI bleeding. Comorbid conditions were present in 5 (28%) patients. Initial hemostasis was achieved endoscopically in 13 patients (72%) and permanent hemostasis in 17 patients (94%). Bleeding recurred in 5 patients (2 with concomitant disease) in the injection group; 3 were successfully retreated by heat probe coagulation and epinephrine injection, 1 with hemoclip application and 1 by surgery. There was no recurrent bleeding in thermal treatment group. Thermal treatment was significantly superior to injection (p = 0.0029). CONCLUSIONS: Endoscopic thermal coagulation with or without epinephrine injection should be the initial treatment of choice for Dieulafoy's lesions. Mortality is lowest in patients with no significant comorbidity and an unremarkable medical history. PMID- 11923768 TI - Endoscopic examination with the duodenoscope at ERCP: frequency of lesions and accuracy of detection. AB - BACKGROUND: Little is known about the yield and accuracy of endoscopic examination with the duodenoscope after ERCP, and there appears to be no uniform standard of practice for endoscopic examination in this setting. METHODS: Over a 1-year period, all patients undergoing ERCP also underwent endoscopic examination of the duodenum, stomach, and esophagus with the duodenoscope upon completion of ERCP. Patients were then placed in the left lateral decubitus position and upper endoscopy was performed with a standard forward-viewing endoscope. The same investigator performed both endoscopic examinations. Patients were queried about prior ulcer disease, upper GI symptoms, and use of antisecretory medication as well as nonsteroidal anti-inflammatory drugs. RESULTS: During the 12-month study, 368 patients (mean age 53 years) underwent 474 procedures. The most frequent indication for ERCP was suspected choledocholithiasis (24%). Endoscopic examination was normal in 42% of patients, and the most frequent endoscopic finding was hiatal hernia (132 patients, 36%). The lesions most frequently missed with the duodenoscope were trauma at the upper esophageal sphincter (12 patients) and small benign gastric polyps (3 patients). Examination with the duodenoscope was superior to the forward-viewing endoscope for identification of Schatzki's ring. No carcinoma, gastric varices, or severe esophagitis were missed with the duodenoscope. CONCLUSIONS: Given the frequency of endoscopic lesions, a yield of positive findings similar to that obtained with the forward-viewing endoscope, and the ease of performing endoscopy with the duodenoscope, endoscopic examination of the esophagus, stomach, and duodenum with the duodenoscope should become standard practice after ERCP. PMID- 11923767 TI - Retroflexed endoscopic band ligation of bleeding internal hemorrhoids. AB - BACKGROUND: Elastic band ligation is a well-established nonoperative method for treatment of bleeding internal hemorrhoids, stages II-III. Usually, one or two bands are placed at a single session by using rigid instruments. The aim of this study was to assess the feasibility, tolerability, safety, and efficacy of multiple band ligation of internal hemorrhoids performed in one session by using a flexible endoscope with an attached band ligation device in the retroflexed position. METHODS: Eighty-three patients with chronically bleeding and/or prolapsing internal hemorrhoids were treated by retroflexed endoscopic band ligation. From 1 to 6 bands were placed in a single session. Bands were targeted at the apex and proximal body of the internal hemorrhoid so that final band placement was entirely proximal to the dentate line. Malpositioned bands were removed by using a novel method. Patients were followed prospectively to assess tolerance, complications, and efficacy. Retreatment was offered if the desired result was not achieved. RESULTS: A mean of 3.0 (SD 1.2) bands (range 1-6) were placed in a single session. Five percent of bands were malpositioned and removed. Patients were followed for 26 (17) months (range 1-52 months). An excellent result was achieved in 80% of patients with stage II hemorrhoids. Patients with stage II hemorrhoids were more likely to have an excellent result compared with patients with stage III hemorrhoids (80% vs. 54%, p < 0.01). Retroflexed endoscopic band ligation was well tolerated overall. The rate of major, nonfatal complications was 4%. CONCLUSIONS: Retroflexed endoscopic band ligation is a feasible, well-tolerated, effective, and safe for treatment of bleeding stage II internal hemorrhoids. A novel method of endoscopic band removal is described. PMID- 11923769 TI - The role of juxtapapillary duodenal diverticula in biliary stone disease. AB - BACKGROUND: Duodenal diverticula are acquired lesions found more often in older patients; when located near the major duodenal papilla they are called juxtapapillary. The prevalence of juxtapapillary duodenal diverticula (JDD) in the general population is around 20%; they are often associated with biliary lithiasis. This study assessed the association between JDD and biliary stone disease, particularly choledocholithiasis. METHODS: Four hundred fifty ERCPs were performed in 420 patients from 1995 through 1999. The results for 300 ERCPs that were performed in 285 patients for suspected biliary lithiasis were analyzed. RESULTS: JDD were present in 21.42% of the 420 patients; most were found in patients in the eighth and ninth decades of life. Patients with JDD had bile duct stones alone more often than patients without JDD (44% vs. 24%). According to their features, these bile duct stones were characterized as mainly primary. The existence of JDD influences bile duct diameter irrespectively of the presence of bile duct stones. CONCLUSIONS: JDD are important causative factors in the formation of bile duct stones. PMID- 11923770 TI - Relationship between age and site of colorectal cancer based on colonoscopy findings. AB - BACKGROUND: Colorectal cancer occurs more frequently in older people. Because the population of aged persons is increasing, a better understanding of the characteristics of colorectal cancer with respect to age would be useful. The purpose of this study was to determine whether there is any relationship between the site of colorectal adenoma and adenocarcinoma in the colon and age. METHODS: Colonoscopy was performed (September 1995 to December 1998) on 2942 consecutive patients (1907 men, 1035 women; mean age 61 years, range 11 to 95 years) with no history of colorectal adenoma, adenocarcinoma, or inflammatory bowel disease. The occurrence of colorectal neoplasia, histologically proven as adenoma or adenocarcinoma, was analyzed for a possible association between site in the colon and patient age. RESULTS: Adenocarcinoma was found in 191 patients (196 lesions). The proportion of patients with right-sided colonic adenocarcinoma increased with patient age: < 50 years, 15% (2/13); 50 to 59 years, 21% (8/39); 60 to 69 years, 32% (18/57); 70 to 79 years, 42% (25/49); > or =80 years, 57% (16/28). The proportion of patients with right-sided adenoma did not significantly differ among age groups: < 50 years, 40% (98/246); 50 to 59 years, 41% (280/678); 60 to 69 years, 46% (459/1001); 70 to 79 years, 53% (270/508); and > or =80 years, 57% (87/152). CONCLUSION: The frequency of right-sided colon cancer increases with patient age. Hence, colonoscopy may be indicated in the elderly for colorectal cancer screening. Over half of colon carcinomas may be missed if sigmoidoscopy alone is used for screening. PMID- 11923771 TI - Practical radiation safety and protection for the endoscopist during ERCP. PMID- 11923773 TI - Recurrent hemorrhage caused by ileal carcinoid. PMID- 11923772 TI - Bleeding jejunal Dieulafoy lesion. PMID- 11923774 TI - The "scarf-ring sign" of ball valve syndrome. PMID- 11923775 TI - Gastric cancer metastatic to the colon. PMID- 11923776 TI - Fluorescein electronic endoscopy: a novel method for detection of early stage gastric cancer not evident to routine endoscopy. AB - BACKGROUND: Fluorescence endoscopy with fluorescein sodium in the stomach was evaluated by using a newly developed fluorescence electronic endoscopic system. METHODS: Sixteen patients with early stage gastric cancer diagnosed by white light endoscopy and chromoendoscopy underwent fluorescein electronic endoscopy before surgery. The resection specimens underwent thorough histopathologic evaluation. RESULTS: About 10 seconds after intravenous injection of fluorescein, fluorescence appeared and immediately spread throughout the gastric surface. A few minutes later, differentiated early stage gastric cancers with more abundant stroma than surrounding normal mucosa exhibited significantly stronger fluorescence, and those with less stroma exhibited weaker fluorescence than the surrounding normal mucosa. Undifferentiated early stage gastric cancers, in which the stroma became wider because foveolae were collapsed from malignant invasion, expressed stronger fluorescence intensity. In all cases, the borders of early stage gastric cancers were clearly demonstrated. Among the 16 patients, 6 accompanying flat lesions and 1 tiny lesion not evident by routine endoscopy were detected. The extent of the cancers, as determined by fluorescence endoscopy, were similar to those determined histopathologically. CONCLUSIONS: Fluorescein electronic endoscopy is useful in determining the extent within the mucosa of gastric cancers when this is obscure by standard endoscopic observation, and for detecting extremely early stage cancer that is not evident by conventional endoscopic observation. PMID- 11923777 TI - 2-Octyl-cyanoacrylate (Dermabond), a new glue for variceal injection therapy: results of a preliminary animal study. AB - BACKGROUND: Endotherapy of bleeding gastric varices is problematic. The aim of this descriptive study in an animal model was to compare 2-octyl-cyanoacrylate (Dermabond), a Food and Drug Administration-approved agent for superficial wound closure, to N-butyl-2-cyanoacrylate (Histoacryl), an agent available outside of the United States for the endoscopic treatment of bleeding gastric varices. METHODS: Eight New Zealand white rabbits were randomly assigned to either the study agent Dermabond or the control agent, Histoacryl. Both active agents were equally mixed with ethiodized poppy seed oil (Ethiodol) and injected into the auricular vein. The animals were euthanized at various time intervals after injection. A 2 cm strip of tissue on either side of the injection site along with the adjacent perivascular tissues were resected for histologic evaluation. RESULTS: The use of 0.5 mL of Dermabond effectively induced vascular occlusion compared with 0.2 mL of Histoacryl. The histologic changes acutely and at 1 week were similar with each mixture. CONCLUSION: Dermabond may be useful in the treatment of gastric variceal bleeding, but further studies are necessary to determine dose response rates in animals and humans. PMID- 11923778 TI - A new technique for endoscopic mucosal resection with an insulated-tip electrosurgical knife improves the completeness of resection of intramucosal gastric neoplasms. AB - BACKGROUND: En bloc resection is optimal for the cure of gastric neoplasms by endoscopic mucosal resection (EMR). A new technique was developed for EMR by using an insulated-tip electrosurgical knife (IT-EMR). This is a report on the clinical application of IT-EMR. METHODS: IT-EMR of 123 gastric tumors was performed in 120 patients. The en bloc resection rate, completeness of resection, and associated complications were evaluated. The local recurrence rate was studied for 90 intramucosal lesions followed for more than 6 months without further treatment. RESULTS: The en bloc resection rate for all lesions was 54% (67/123 lesions). The en bloc resection rates were 82% (27/33) for lesions 10 mm or less in size, 54% (29/54) for those between 11 mm and 20 mm, and 31% (11/36) for those of over 20 mm. Complete resection rates in the cases with en bloc resection were 78% (21/27) for lesions 10 mm or less in size, 76% (22/29) for those between 11 mm and 20 mm, and 73% (8/11) for those over 20 mm. There were no episodes of major bleeding that required blood transfusion or surgical intervention; minor bleeding including oozing occurred in 38% (47/123). Perforation occurred in 1 case (1/123; 0.8%). The local recurrence rate for lesions resected en bloc was significantly lower than that for lesions resected as multiple fragments (respectively, 2/49; 4.1% vs. 7/41; 17%: p = 0.041). CONCLUSIONS: IT-EMR is feasible in clinical practice and has a high en bloc resection rate. En bloc resection may reduce the rate of local recurrence. PMID- 11923779 TI - Persimmon bezoars: a successful combined therapy. AB - BACKGROUND: Persimmon phytobezoar, although an infrequent entity, is not rare in some countries. Because of their particular features, management of diospyrobezoars is difficult. A number of surgical, endoscopic, and pharmacologic treatments have been proposed with variable success. This is a description of our experience with 10 patients with a new combination therapy. METHODS: Ten patients (7 men, 3 women; mean age 46.4 years) were treated with a new fragmentation technique consisting of a saw-like effect with a large polypectomy snare followed by administration of cellulase, cysteine, and metoclopramide. RESULTS: Resolution was achieved in 8 patients. Complications (intestinal obstruction) developed in 2 patients. CONCLUSIONS: Our combined therapy is safe and highly efficacious but caution must be exercised because intestinal obstruction can occur if large fragments pass through the pylorus. A more extensive study is required to assess these clinical observations. PMID- 11923780 TI - Endoscopic and histologic features of sodium phosphate bowel preparation-induced colonic ulceration: case report and review. PMID- 11923781 TI - Lymphoplasmacytic sclerosing pancreatitis and cholangitis. PMID- 11923782 TI - Diffuse esophageal leiomyomatosis: case report and review. PMID- 11923783 TI - Preoperative localization of a neuroendocrine tumor of the pancreas with EUS guided fine needle tattooing. PMID- 11923784 TI - Gastric arteriovenous malformation: Doppler EUS-guided diagnosis and therapy. PMID- 11923785 TI - Spontaneous resolution of multiple fundic gland polyps after cessation of treatment with lansoprazole and Nissen fundoplication: a case report. PMID- 11923786 TI - Endoscopic management of afferent loop syndrome of malignant etiology. PMID- 11923787 TI - Duodenal nodular lymphoid hyperplasia caused by giardiasis infection in a patient who is immunodeficient. PMID- 11923788 TI - Jejunal perforation complicating PEG insertion in a child. PMID- 11923789 TI - The future of simulators in GI endoscopy: an unlikely possibility or a virtual reality? PMID- 11923790 TI - Meta-analysis, part I. PMID- 11923791 TI - Renal artery stenosis: a cardiovascular perspective. AB - BACKGROUND: Renal artery stenosis (RAS) is an important clinical entity that can lead to uncontrolled hypertension and progressive renal failure. The most common causes of RAS are atherosclerosis and fibromuscular dysplasia. Because the diagnosis of renovascular hypertension is established only when revascularization of a stenosed renal artery results in cure or improvement in patients hypertension, establishment of a causal relationship between RAS and hypertension continues to present a challenge. Therefore, a high index of suspicion is essential in the diagnosis of this condition. METHODS: Multiple tests, both invasive and noninvasive, are available as screening methods. Angiography remains the gold standard for diagnosis of RAS. Besides its value in establishing the diagnosis, it provides anatomic information regarding the site and severity of stenoses and appropriate revascularization strategies. Magnetic resonance angiography and duplex ultrasonography are the most promising and accurate noninvasive screening tests available, even in the presence of renal insufficiency. With advances in percutaneous transluminal angioplasty techniques, including renal artery stenting, many more patients are eligible for less invasive and effective revascularization strategies compared with the traditional surgical procedures. RESULTS: Revascularization of a stenosed renal artery is associated with preservation of renal function and better control of hypertension, unstable angina, and congestive heart failure. Because atherosclerotic RAS is associated with generalized atherosclerosis, aggressive risk factor modification and antiplatelet therapy are integral in the management of RAS regardless of the revascularization strategy. PMID- 11923792 TI - Cost and cost-effectiveness studies in heart failure research. AB - BACKGROUND: Heart failure is a major and increasing cause of death and disability and accounts for significant resource use. In the United States alone, the prevalence is 4.6 million, with an incidence rate of 550,000 new cases a year and approximately 957,000 hospitalizations a year. METHODS AND RESULTS: Methods of evaluating cost and outcome and of comparing cost with outcome are reviewed. Economic and cost-effectiveness studies in heart failure research, especially those related to clinical trials, are reviewed in the therapeutic areas of digoxin, angiotension-converting enzyme inhibition, beta blockers, disease management, and transplantation. CONCLUSION: In an era in which economic constraints on medical resource use limit the ability to give all services to all patients, economic studies can help guide more rational decision making. Economic studies in heart failure can be expected to improve and so help society to make better, more informed choices. PMID- 11923793 TI - Study of the Therapeutic Effects of Intercessory Prayer (STEP): study design and research methods. AB - BACKGROUND: The effect of intercessory prayer (IP) on outcome in cardiac cases has been evaluated previously, but results are controversial. The goals of the Study of the Therapeutic Effects of Intercessory Prayer (STEP) are to evaluate the effects of receipt of additional study IP and awareness of receipt of additional study IP on outcomes in patients undergoing coronary artery bypass graft surgery. STEP is not designed to determine whether God exists or whether God does or does not respond to IP. METHODS: STEP is a multicenter, controlled trial of 1802 patients in 6 US hospitals, randomized to 1 of 3 groups. Two groups were informed that they may or may not receive 14 consecutive days of additional IP starting the night before coronary artery bypass graft surgery; Group 1 received IP, Group 2 did not. A third group (Group 3) was informed that they would receive additional IP and did so. Three mainstream religious sites provided daily IP for patients assigned to receive IP. At each hospital, research nurses blinded to patient group assignment reviewed medical records to determine whether complications occurred, on the basis of the Society for Thoracic Surgeons definitions. A blinded nurse auditor from the Coordinating Center reviewed every study patient's data against the medical record before release of study forms. RESULTS: The STEP Data and Safety Monitoring Board reviewed patient safety and outcomes in the first 900 study patients. Patients were enrolled in STEP from January 1998 to November 2000. PMID- 11923794 TI - Pharmacodynamic profile of the direct thrombin antagonist bivalirudin given in combination with the glycoprotein IIb/IIIa antagonist eptifibatide. AB - BACKGROUND: Because of the adverse characteristics associated with heparin, direct antagonists of thrombin have been investigated as anticoagulants during percutaneous coronary interventions. However, the hematologic and clinical interactions between direct thrombin antagonists and inhibitors of platelet glycoprotein IIb-IIIa are incompletely explored. METHODS: Forty-two patients who underwent elective percutaneous coronary intervention were randomized to receive a bivalirudin 1.0 mg/kg bolus followed by a 4-hour infusion at 2.5 mg/kg/h; a bivalirudin 0.75 mg/kg bolus followed by a 4-hour infusion at 1.75 mg/kg; or a heparin 60 U/kg bolus. All the patients also received eptifibatide, given as 2 sequential boluses of 180 microg/kg followed by a 2 microg/kg/min infusion for 18 to 24 hours, and aspirin. RESULTS: After the bolus dose of the study drug, turbidimetric platelet aggregation in response to 5 micromol/L adenosine diphosphate increased in patients assigned to heparin but not those assigned to bivalirudin. After eptifibatide, platelet aggregation was eliminated in all 3 treatment groups. The effect of heparin and the effects of both bivalirudin regimens on the formation of thrombin antithrombin complexes and prothrombin fragment 1.2 were comparable. Neither agent affected the formation of platelet monocyte complexes or expression of CD 63 lysosomal antigen. There were no major bleeding events, and a single non-Q-wave myocardial infarction (MI) occurred in a patient treated with bivalirudin. CONCLUSION: These findings show the feasibility of combining the direct thrombin antagonist bivalirudin with a potent antagonist of platelet glycoprotein IIb-IIIa. Clinical trials are needed to assess the safety and efficacy of this combination. PMID- 11923795 TI - Glycoprotein IIb-IIIa inhibition with abciximab and postprocedural risk assessment: lessons from the evaluation of platelet IIb/IIIa inhibitor for stenting trial and implication for ad hoc use of glycoprotein IIb-IIIa antagonists. AB - BACKGROUND: Angiographic features of vessels in which stents have been deployed can be used to predict the risk of postprocedural ischemic events. The purpose of this study was to compare the effects of abciximab in patients with and without high-risk postprocedure features. METHODS AND RESULTS: Protocol-mandated stent implantation was performed in 1586 patients in the Evaluation of Platelet IIb/IIIa Inhibitor for Stenting trial, 783 of whom received abciximab, and was successful in 97% of the patients. High-risk features were defined as the presence of Thrombolysis In Myocardial Infarction (TIMI) flow <3, presence of thrombus or major dissection, or residual stenosis >10%. The primary endpoint was a composite of death, myocardial infarction, and urgent target vessel revascularization at 30 days. High-risk features were present in 21% of the patients. In patients without high-risk features after stent placement, abciximab reduced the primary endpoint from 9.0% to 3.9% (P <.001) compared with 16.2% to 8.6% (P =.046) in patients in whom high-risk features were present. There was no statistical evidence of interaction between abciximab treatment and the presence or absence of high-risk features. CONCLUSION: Glycoprotein IIb-IIIa antagonism with abciximab is equally effective in prevention of a composite of ischemic events in patients with and without high-risk features after stent placement. However, patients in whom high-risk features are present after stent placement are at increased risk of ischemic cardiac events even with abciximab treatment. PMID- 11923796 TI - The Ultegra rapid platelet-function assay: comparison to standard platelet function assays in patients undergoing percutaneous coronary intervention with abciximab therapy. AB - BACKGROUND: Despite the proven benefit of glycoprotein IIb/IIIa inhibitors during percutaneous coronary intervention, significant interpatient variability exists in antiplatelet response. Furthermore, a diminished degree of platelet inhibition is an independent predictor of adverse cardiac events, highlighting the need for accurate and precise monitoring of platelet function. METHODS: Patients (n = 192) who underwent elective percutaneous coronary intervention at 4 centers were enrolled. The following 3 time points were studied: 1, baseline, before abciximab bolus administration; 2, during, within 1 hour of abciximab bolus administration; and 3, post, 24 hours after abciximab bolus administration or at the time of patient discharge, whichever occurred first. The following 3 assays were compared at all time points: Ultegra rapid platelet-function assay (Ultegra RPFA), conventional turbidometric platelet aggregometry, and receptor binding assay with [125I]-abciximab. Variability in Ultegra RPFA measurements between operators was determined with performance of the assays at the point of care and in the laboratory. A sub-study of 22 patients at 1 center was performed in which the laboratory scientist performed all 3 assays in duplicate at each time point. RESULTS: Comparison with the receptor binding assay and conventional platelet aggregometry in 120 patients showed that the Ultegra RPFA correlated with aggregometry (r = 0.89) and with the receptor binding assay (r = 0.89). There was good agreement (r = 0.80) between values obtained by intended users and those obtained by laboratory scientists. Furthermore, Ultegra RPFA values had equivalent precision to the standard assays. CONCLUSION: The Ultegra RPFA has equivalent accuracy and precision when compared with the 2 reference assays studied. Ultegra RPFA measurements are not operator-dependent and are not influenced by concomitant medications, hematologic parameters, or demographics. PMID- 11923797 TI - Assessment of the safety and efficacy of the DUETT vascular hemostasis device: final results of the safe and effective vascular hemostasis (SEAL) trial. AB - OBJECTIVE: We sought to determine the safety and efficacy of the novel DUETT vascular hemostasis device in comparison with standard manual compression after diagnostic and interventional coronary procedures. BACKGROUND: Vascular hemostasis devices are increasingly used to improve patient comfort and speed mobilization after coronary and peripheral vascular procedures. Currently available devices have certain limitations, however. METHODS: At 16 clinical sites, 630 patients who underwent diagnostic or interventional coronary procedures were randomized 5:3 to the DUETT sealing device or standard manual compression. The primary study end points were time to hemostasis and ambulation and the incidence of major vascular complications at 30 days. RESULTS: Time to hemostasis from the completion of the procedure (catheter removal; median) was 14 minutes (interquartile range [IQR], 10, 17 minutes) in the DUETT group and 195 minutes (IQR, 46, 351 minutes) in the standard compression group (P <.001), and time from sheath removal (median) was 7 minutes (IQR, 6, 8 minutes) and 20 minutes (IQR, 15, 30 minutes) for the 2 groups, respectively (P <.001). Time to ambulation from catheter removal (median) was 338 minutes (IQR, 223, 526 minutes) in the DUETT group and 705 minutes (IQR, 400, 1120 minutes) in the standard compression group (P <.001). Major complications occurred in 3.6% of the DUETT group and 1.7% of the standard compression group (P =.22), with a diminishing risk of complications in the DUETT group as experience was accrued. Similar benefits from DUETT use were seen in patients who underwent both diagnostic and interventional procedures. CONCLUSION: The DUETT sealing device allows immediate arterial sheath removal after both diagnostic and interventional procedures, dramatically reducing time to hemostasis and patient ambulation without compromising patient safety in comparison with standard compression techniques. PMID- 11923798 TI - Coronary stent implantation in patients older than 75 years of age: clinical profile and initial and long-term (3 years) outcome. AB - OBJECTIVE: The objective of this study was to compare the initial and long-term outcome of elderly and younger patients after coronary stent implantation. METHODS: The evolutions of 76 patients aged >75 years and of 860 patients aged < or =75 years who underwent consecutive stenting (from June 1991 to June 1997) were compared in a cohort study. RESULTS: The elderly patients had lower left ventricular ejection fractions (0.58 +/- 0.14 vs 0.61 +/- 0.13; P =.03) and more frequently had unstable angina (78.9% vs 55.3%; P <.0001), previous heart failure (10.5% vs 4.9%; P =.03), and multivessel disease (68.4% vs 58.3%; P =.08). After the procedure, the elderly patients showed a higher inhospital mortality rate (6.6% vs 2.4%; P =.03) and myocardial infarction rate (5.3% vs 1.7%; P =.04). The long-term follow-up period (mean, 3.2 +/- 1.4 years; median, 3.0 years) showed in the elderly a higher mortality rate (15.4% vs 5.8%; P =.006), a lower rate of repeat revascularization (9.2% vs 19.7%; P =.04), and a similar incidence rate of major adverse cardiac events (27.7% vs 28.2%; P =.93). Multivariate analysis of the elderly group identified female gender (hazard ratio, 2.19; 95% CI, 1.18 to 4.06; P =.012) and presence of multivessel disease (hazard ratio, 2.35; 95% CI, 1.05 to 5.26; P =.037) as independent predictors of further events. CONCLUSION: Patients aged >75 years have a less favorable baseline profile and higher inhospital and 3-year mortality rates. However, the incidence rate of major adverse cardiac events in the long term is acceptable and similar to that of younger patients. PMID- 11923799 TI - Natural history of small and medium-sized side branches after coronary stent implantation. AB - OBJECTIVE: Our purpose was to identify angiographic and procedural predictors for acute and late side branch occlusion after coronary stent implantation. METHODS: We evaluated 185 patients with 185 lesions with 255 side branches with a mean reference diameter of 1.45 +/- 0.38 mm; the lesions were covered by 240 stents. Angiographic follow-up was completed in 99 patients with 133 side branches 206 +/ 120 days after stent implantation and clinical follow-up was available in 136 patients. Side branch occlusion (SBO) was defined as a Thrombolysis In Myocardial Infarction (TIMI) flow < or =1. RESULTS: Acute SBO affected 54 side branches in 49 patients and was not associated with death or Q-wave infarction. By logistic regression, independent predictors for acute SBO were (1) the reference side branch diameter (RLD) at baseline (OR [odds ratio] 0.217, 95% CI 0.07-0.67, P =.008); (2) an ostial side branch stenosis before stenting (OR 2.96, 95% CI 1.26 6.95, P =.013); (3) the involvement of the side branch origin within the lesion of the parent vessel (OR 2.77, 95% CI 1.17-6.57, P =.021); and (4) the balloon-to artery ratio (OR 4.66, 95% CI 1.18-18.42, P =.028). Among the initially occluded side branches, 81.8% were spontaneously reperfused at follow-up. Late SBO involved 12% of the side branches without impaired antegrade flow after stenting and was predicted by the initial RLD of the side branch (OR 0.07, 95% CI 0.01 0.8, P =.032). Chronic SBO occurred in 13.5% of cases and was also predicted by the baseline RLD (OR 0.13, 95% CI 0.02-0.8, P =.028). CONCLUSIONS: Acute SBO after stenting occurred in 21.2% of cases and had a benign course. Most acutely occluded side branches underwent late spontaneous reperfusion. A baseline side branch diameter >1.4 mm predicted a preserved antegrade flow immediately after stent implantation, as well as during follow-up. PMID- 11923800 TI - Expanding the outcomes in clinical trials of heart failure: the quality of life and economic components of EPHESUS (EPlerenone's neuroHormonal Efficacy and SUrvival Study). AB - BACKGROUND: Although quantification of the impact of treatment on survival remains the cornerstone of clinical research, the assessment of a broader range of disease outcomes is growing increasingly important. This manuscript provides an overview of the considerations made regarding quantification of a range of clinical and economic outcomes in the EPHESUS (EPlerenone's neuroHormonal Efficacy and SUrvival Study) study, a 6200-patient, randomized, controlled trial of aldosterone blockade in patients with heart failure as a complication of acute myocardial infarction. METHODS AND RESULTS: Relevant clinical outcomes include disease progression (mortality, hospitalization), health status (symptoms, functioning, and quality of life), and costs. The primary hypothesis for the quality of life component of EPHESUS is that eplerenone will improve health status as defined by the Kansas City Cardiomyopathy Questionnaire summary score. In addition to the Kansas City Cardiomyopathy Questionnaire, the Short Form-12, the EuroQOL, the Medical Outcomes Study-Depression Scale and an Anxiety measure will be collected throughout the trial. Health care resource utilization including hospitalizations, emergency room visits, outpatient procedures and tests, and medications will also be collected. Analyses will estimate and describe the effect of aldosterone blockade on costs over time. If both the clinical effect and costs are greater for patients receiving aldosterone blockade, then eplerenone's cost-effectiveness will be estimated. CONCLUSION: The EPHESUS trial has been designed to quantify a wide range of clinical outcomes. This broad range of data will allow a comprehensive assessment of the potential benefits of aldosterone blockade on patient health status and costs. PMID- 11923801 TI - Antiarrhythmic effects of azimilide in paroxysmal supraventricular tachycardia: efficacy and dose-response. AB - BACKGROUND: Azimilide is a novel classification III antiarrhythmic agent that blocks both I(Kr)and I(Ks)and shows evidence of efficacy in patients with atrial fibrillation or flutter. Its effect on paroxysmal supraventricular tachycardia (PSVT) has not been reported. METHODS AND RESULTS: One hundred ninety-three patients with symptomatic PSVT were enrolled in 4 double-blind, randomized, placebo-controlled clinical trials with almost identical trial design (supraventricular arrhythmia-1 [SVA-1], SVA-2, SVA-3, and SVA-4), performed as a separate stratum of studies that also included a stratum of patients with atrial fibrillation or flutter. Patients received oral azimilide (100 mg in SVA-1 and SVA-3, 35 or 75 mg in SVA-2, and 125 mg in SVA-4) or matching placebo twice daily for 3 days (loading period), followed by once-daily dosing (maintenance period). The primary outcome variable was the first recording of a symptomatic supraventricular arrhythmia (either PSVT, atrial fibrillation, or atrial flutter) recorded on a transtelephonic electrocardiographic event recorder. The duration of study was 270 days in SVA-1 and SVA-2 and 180 days in SVA-3 and SVA-4. Combined analysis results of the PSVT stratum from the 4 studies showed a dose response relationship in prolongation of time to recurrence (P =.02). In the combined 100-mg daily dose, the hazard ratio (placebo:azimilide) was 2.35 (P =.023). The hazard ratio for the 125-mg daily dose measured 1.28 (P = not significant). However, the time to recurrence was prolonged when the patients receiving 100 and 125 mg daily were combined and compared with placebo (P =.02). There were no deaths and 1 case of torsades de pointes. CONCLUSION: These trial results suggest a significant dose-related suppression of PSVT with azimilide, with a low risk of serious adverse events. PMID- 11923802 TI - Comparison of treadmill scores with physician estimates of diagnosis and prognosis in patients with coronary artery disease. AB - OBJECTIVE: Our purpose was to compare exercise test scores and ST measurements with a physician's estimation of the probability of the presence and severity of angiographic disease and the risk of death. The American College of Cardiology/American Heart Association exercise testing guidelines provide equations to calculate treadmill scores and recommend their use to improve the predictive accuracy of the standard exercise test. However, if physicians can estimate the probability of coronary artery disease and prognosis as well as the scores, there is no reason to add this complexity to test interpretation. METHODS: A clinical exercise test was performed and an angiographic database was used to print patient summaries and treadmill reports. The clinical/treadmill test reports were sent to expert cardiologists and to 2 other groups, including randomly selected cardiologists and internists. They classified the patients summarized in the reports as having a high, low, or intermediate probability for the presence of any severe angiographic disease and estimated a numerical probability from 0% to 100%. The Social Security Death Index was used to determine survival status of the patients. RESULTS: Twenty-six percent of the patients had severe angiographic disease, and the annual mortality rate for the population was 2%. Forty-five expert cardiologists returned estimates on 473 patients, 37 randomly chosen practicing cardiologists returned estimates on 202 patients, 29 randomly chosen practicing internists returned estimates on 162 patients, 13 academic cardiologists returned estimates on 145 patients, and 27 academic internists returned estimates on 272 patients. When probability estimates for presence and severity of angiographic disease were compared, in general, the treadmill scores were superior to physicians' and ST analysis at predicting severe angiographic disease. When prognosis was estimated, treadmill prognostic scores did as well as expert cardiologists and better than most other physician groups. CONCLUSION: Estimates of the presence of clinically significant and severe angiographic coronary artery disease provided by scores were superior to physician estimates and ST analysis alone. Estimates of prognosis provided by scores were similar to the estimates made by expert cardiologists and more accurate than the estimates made by most other physician groups. PMID- 11923803 TI - Comparison of 2-dimensional echocardiography and myocardial perfusion imaging for diagnosing myocardial infarction in emergency department patients. AB - BACKGROUND: Both 2-dimensional echocardiography and myocardial perfusion imaging (MPI) with technetium-99m based agents have been used to identify patients in the emergency department with myocardial infarction (MI). However, the inclusion of small numbers of patients in prior studies limits the accurate assessment of sensitivity of the 2 techniques. METHODS: Gated MPI was used as part of the initial triage process in patients initially considered at low to moderate risk for acute coronary syndromes (no ST elevation or depression). Patients diagnosed with MI also underwent echocardiography. MPI results were considered positive if there was a perfusion defect associated with abnormal wall motion or thickening, and echocardiographic results were considered positive if there were segmental wall motion abnormalities or ejection fraction of less than 40%. RESULTS: Both tests were performed on 141 patients. The sensitivities for MI for echocardiography (91%; 95% CI, 86%-95%) and MPI (89%; 95% CI, 83%-94%) were similar. Patients who had either negative echocardiographic results (peak creatine kinase level [CK], 325 +/- 206 vs 582 +/- 614 U/L; P =.003) or negative MPI results (peak CK, 313 +/- 227 vs 590 +/- 620 U/L; P =.001) had smaller MIs as estimated with peak CK values. Ejection fraction was highly correlated between the 2 techniques (r = 0.82; P <.001). CONCLUSION: Both echocardiography and MPI have a high sensitivity for identifying patients in the emergency department who have MI. False negative studies with either technique were associated with small MIs. PMID- 11923804 TI - Usefulness of an echo-contrast agent for assessment of coronary flow velocity and coronary flow velocity reserve in the left anterior descending coronary artery with transthoracic doppler scan echocardiography. AB - BACKGROUND: This study assessed the feasibility of transthoracic Doppler scan echocardiography (TTDE) combined with echo-contrast agent in measuring coronary flow velocity (CFV) and coronary flow velocity reserve (CFVR) in the left anterior descending artery. METHODS: In 68 consecutive patients who underwent cardiac angiography, TTDE was recorded before and after induction of a hyperemic condition with intravenous administration of adenosine triphosphate (0.14 mg/kg/min). After CFV values returned to baseline, the same measurements were repeated while an echo-contrast agent was continuously infused. CFVR was assessed as the ratio of hyperemic to basal CFV. The pulsed wave Doppler scan quality was graded from 1 to 3 (TTDE score: 1, no signal detection; 2, poor definition of outline; 3, optimal outline definition). RESULTS: Before enhancement, CFVR could not be measured in 20 patients. Prevalence of delayed stenosis (Thrombolysis In Myocardial Infarction [TIMI] II grade flow) in these patients (30%) was significantly greater than in those whose CFVRs could be measured without enhancement (2%; P <.01). TTDE scores both at baseline and after hyperemia were significantly improved with contrast enhancement (before, 2.8 +/- 0.6; after, 3.0 +/- 0.3; P <.01; before, 2.6 +/- 0.7; after, 3.0 +/- 0.3; P <.01; respectively). Overall contrast enhancement increased the rate of successful CFVR measurement from 70% to 97% (P <.01). Sensitivity and specificity of significant left anterior descending artery stenosis detection with CFVR of <2.0 were 94.4% and 87.8%, respectively. CONCLUSION: These data suggest that administration of echo contrast agent improves pulse wave Doppler scan quality and thus the feasibility of measuring CFVR. PMID- 11923805 TI - Effects of bisoprolol fumarate on left ventricular size, function, and exercise capacity in patients with heart failure: analysis with magnetic resonance myocardial tagging. AB - BACKGROUND: Recent data suggest that beta-blockers can be beneficial in subgroups of patients with chronic heart failure (CHF). For metoprolol and carvedilol, an increase in ejection fraction has been shown and favorable effects on the myocardial remodeling process have been reported in some studies. We examined the effects of bisoprolol fumarate on exercise capacity and left ventricular volume with magnetic resonance imaging (MRI) and applied a novel high-resolution MRI tagging technique to determine myocardial rotation and relaxation velocity. METHODS: Twenty-eight patients (mean age, 57 +/- 11 years; mean ejection fraction, 26 +/- 6%) were randomized to bisoprolol fumarate (n = 13) or to placebo therapy (n = 15). The dosage of the drugs was titrated to match that of the the Cardiac Insufficiency Bisoprolol Study protocol. Hemodynamic and gas exchange responses to exercise, MRI measurements of left ventricular end-systolic and end-diastolic volumes and ejection fraction, and left ventricular rotation and relaxation velocities were measured before the administration of the drug and 6 and 12 months later. RESULTS: After 1 year, heart rate was reduced in the bisoprolol fumarate group both at rest (81 +/- 12 before therapy versus 61 +/- 11 after therapy; P <.01) and peak exercise (144 +/- 20 before therapy versus 127 +/ 17 after therapy; P <.01), which indicated a reduction in sympathetic drive. No differences were observed in heart rate responses in the placebo group. No differences were observed within or between groups in peak oxygen uptake, although work rate achieved was higher (117.9 +/- 36 watts versus 146.1 +/- 33 watts; P <.05) and exercise time tended to be higher (9.1 +/- 1.7 minutes versus 11.4 +/- 2.8 minutes; P =.06) in the bisoprolol fumarate group. A trend for a reduction in left ventricular end-diastolic volume (-54 mL) and left ventricular end-systolic volume (-62 mL) in the bisoprolol fumarate group occurred after 1 year. Ejection fraction was higher in the bisoprolol fumarate group (25.0 +/- 7 versus 36.2 +/- 9%; P <.05), and the placebo group remained unchanged. Most changes in volume and ejection fraction occurred during the latter 6 months of treatment. With myocardial tagging, insignificant reductions in left ventricular rotation velocity were observed in both groups, whereas relaxation velocity was reduced only after bisoprolol fumarate therapy (by 39%; P <.05). CONCLUSION: One year of bisoprolol fumarate therapy resulted in an improvement in exercise capacity, showed trends for reductions in end-diastolic and end-systolic volumes, increased ejection fraction, and significantly reduced relaxation velocity. Although these results generally confirm the beneficial effects of beta-blockade in patients with chronic heart failure, they show differential effects on systolic and diastolic function. PMID- 11923806 TI - Endogenous nitric oxide prevents myocardial ischemia in patients with hypertension and left ventricular hypertrophy. AB - BACKGROUND: Left ventricular hypertrophy (LVH) caused by chronic pressure overload is associated with increased risk of myocardial ischemia without epicardial coronary artery disease. We aimed to test the hypothesis that endogenous nitric oxide (NO) prevents myocardial ischemia in patients with LVH. METHODS: Epicardial coronary blood flow (Doppler wire and quantitative coronary arteriography) and myocardial lactate metabolism (paired arterial and coronary sinus blood sampling) were measured in 12 patients with hypertension, LVH, and angiographically normal epicardial coronary arteries and in 7 control subjects. Measurements were done under 3 pacing protocols: with no treatment (control), with intracoronary N(G)-monomethyl-L-arginine (L-NMMA; NO synthesis inhibitor), and with intracoronary L-arginine (NO substrate). RESULTS: In control subjects the myocardial lactate extraction ratio was normal and stable during the 3 pacing protocols. In contrast, lactate uptake was significantly decreased from 0.21 +/- 0.05 to 0.10 +/- 0.06 (P <.05) during L-NMMA pacing in patients with LVH; in 6 of them, lactate production was demonstrated. After L-arginine administration, the lactate extraction ratio during pacing was normalized (0.18 +/- 0.04) and lactate production was not observed in any patient. The level of myocardial lactate uptake at peak pacing after L-NMMA was correlated with that under untreated condition (P <.0001). CONCLUSIONS: In patients with hypertension, LVH, and angiographically normal coronary arteries, inhibition of endogenous NO synthesis in the coronary circulation unmasked myocardial ischemia during tachycardia, and L-arginine reversed the adverse effects of L-NMMA. Although the precise mechanism remains to be determined, our results suggest that constitutive NO in the coronary circulation plays an anti-ischemic role in this population. PMID- 11923807 TI - Septal wall thinning and systolic dysfunction in patients with hypertrophic cardiomyopathy caused by a cardiac troponin I gene mutation. AB - BACKGROUND: Lysine 183 deletion in the cardiac troponin I gene is 1 of the mutations that causes hypertrophic cardiomyopathy (HCM). However, the clinical course and determinants of poor prognosis in patients with this mutation have not been well established. METHODS AND RESULTS: We analyzed 10 probands with HCM caused by this mutation and their family members. Forty-six of these 79 subjects were found to be carriers, and 33 were non-carriers. All non-carriers had a percent fractional shortening (%FS) of >25% at all ages. By contrast, 7 of 24 carriers >40 years of age had a %FS of <25%, and no carriers <40 years of age had a %FS of <25%. The change in interventricular septal thickness and the change in %FS were significantly correlated (R = 0.758; P =.0017). CONCLUSION: These results suggest that about 30% of patients with HCM caused by a lysine 183 deletion mutation in the cardiac troponin I gene have systolic dysfunction develop after 40 years of age, and that patients with this mutation whose interventricular septal thickness shows a serial decrease should be followed-up closely for development of systolic dysfunction. PMID- 11923808 TI - Left ventricular remodeling in the first year after acute myocardial infarction and the predictive value of N-terminal pro brain natriuretic peptide. AB - BACKGROUND: Left ventricular (LV) remodeling after myocardial infarction (MI) has received much attention because of its severe impact on morbidity and mortality rates. However, the incidence and extent of LV remodeling in a modern infarct population who were offered antiremodeling treatment in compliance with daily clinical practice is unknown. The purpose of this study was to clarify this issue and to evaluate the predictive value of N-terminal pro brain natriuretic peptide (NT-proBNP). METHODS: Forty-two patients with a first transmural MI were examined after 1 week, 1 month, 3 months, 6 months, and 1 year with blood samples and magnetic resonance imaging. RESULTS: In 12 patients (29%), LV end-diastolic volume index (LVEDVI) and LV end-systolic volume index (LVESVI) increased by 24% and 22% (P <.0001; P =.01). In 12 patients (29%), LVEDVI and LVESVI decreased by 19% and 23% (P <.0001; P =.0005), whereas the remaining 18 patients (43%) had stable conditions regarding these LV measures. LV ejection fraction at baseline was significantly reduced in all patient categories but was unchanged over time. Elevated NT-proBNP level at baseline was identified as an independent predictor of increase in LVEDVI during follow-up examination (P =.007). A baseline level of NT-proBNP >115 pmol/L identified patients who later had LV dilatation develop with a sensitivity and specificity of 89% and 68% (area under curve = 0.77). CONCLUSION: In this 1-year follow-up study of patients with a first transmural MI, approximately 30% had significant increments develop in LVEDVI and LVESVI, and LV ejection fraction remained unchanged. Patients in whom LV dilatation developed could be identified early after the MI with elevated plasma levels of NT-proBNP. PMID- 11923809 TI - Effects of sildenafil citrate (viagra) on cardiac repolarization and on autonomic control in subjects with chronic heart failure. AB - BACKGROUND: Cases of sudden death associated with sildenafil citrate use have been reported in men with coronary artery disease. The aim of this study was to investigate the drug's effect on cardiac repolarization and sinus autonomic and vascular control in men with mild chronic heart failure (CHF; New York Heart Association classification II). Changes in these variables could predispose patients to malignant ventricular arrhythmias. METHOD: We measured QT dispersion, the QT-RR slope, and the index of QT variability (QTVI) and analyzed spectral power of RR and systolic blood pressure variability in 10 men with dilated cardiomyopathy and in 10 control subjects after administration of a single 50-mg oral dose of sildenafil citrate or placebo at rest (not followed with any attempt at intercourse). RESULTS: In both groups, oral sildenafil citrate decreased the systolic blood pressure (P <.05) and increased the heart rate (P <.05). In subjects with CHF, it also increased the QT-RR (P <.001) and QTVI (from -0.45 +/- 0.07 to -0.27 +/- 0.07; P <.001), but in controls, it increased the QTVI (from 1.20 +/- 0.08 to -0.78 +/-.014; P <.001). In these subjects and controls, oral sildenafil citrate induced a significant reduction in high frequency, expressed in absolute power (subjects with CHF: from 4.04 +/- 0.14 to 3.43 +/- 0.16 natural logarithm ms2; P <.001; controls: from 5.61 +/- 0.44 to 4.98 +/- 0.32 natural logarithm ms2; P <.05) and in normalized units (P <.05). In subjects with CHF but not in controls, it also significantly increased the low frequency to high frequency ratio (from 1.3 +/- 0.12 to 1.89 +/- 0.16; P <.001) and low frequency expressed in normalized units (P <.05). Sildenafil citrate caused no significant changes in the QT interval or dispersion. CONCLUSION: These findings indicate that, in men with heart failure, sildenafil citrate reduces vagal modulation and increases sympathetic modulation, probably through its reflex vasodilatory action. The autonomic system changes induced with sildenafil citrate could alter QT dynamics. Both changes could favor the onset of lethal ventricular arrhythmias. At the dose usually taken for erectile dysfunction, sildenafil citrate has no direct effect on cardiac repolarization (QT interval or dispersion). PMID- 11923811 TI - Effects of thyroid replacement therapy on arterial blood pressure in patients with hypertension and hypothyroidism. AB - BACKGROUND: Hypothyroidism is frequently accompanied by cardiac dysfunction, increased vascular resistance, and a greater prevalence of hypertension. Treatment of hypothyroidism may lead to normalization of blood pressure, although some patients may exhibit sustained hypertension. The mechanism of this condition may be the alterations in aortic stiffness. METHODS: Aortic stiffness was studied in 30 patients who never received treatment for hypertension or hypothyroidism, 15 patients with normal blood pressure and hypothyroidism, and 15 patients with hypertension and normal thyroid function. Thirty healthy age- and sex-matched subjects with normal thyroid function served as control subjects. Aortic diameter evaluated by M-mode echocardiography and blood pressure measured by a sphygmomanometer were used to calculate aortic stiffness index. RESULTS: Patients with high blood pressure and hypothyroidism, those with normal blood pressure and hypothyroidism, and those with hypertension and normal thyroid function showed increased aortic stiffness index (18.8 +/- 6.4, 11.7 +/- 3.5, and 19.2 +/- 5.3 vs 9.5 +/- 2.7; P <.001) compared with control subjects. In 15 patients with hypertension and hypothyroidism, levothyroxine therapy showed only a small decrease in blood pressure (151/105 +/- 9/9 mm Hg, group A). The remaining 15 patients showed complete normalization of blood pressure (118/83 +/- 8/3 mm Hg, group B). Aortic stiffness index was increased in group A compared with group B both before and after treatment (before, 24.0 +/- 4.1 vs 13.7 +/- 3.2; and after, 22.3 +/- 4.2 vs 11.1 +/- 2.9; P <.001 for both comparisons). Felodipine was given to patients in group A after levothyroxine was administered, resulting in normalization of blood pressure and a significant decrease of aortic stiffness index (P <.001). Aortic stiffness index was decreased in patients with hypothyroidism and hypertension after administration of levothyroxine (9.5 +/- 2.2; P <.001) and felodipine (14.5 +/- 7.5; P <.001) therapy, respectively. Percent changes in systolic blood pressure showed a significant correlation with percent changes in aortic stiffness index in all patients (r = 0.65, P <.001). After multivariate adjustment, aortic stiffness index (odds ratio = 1.9932; confidence interval = 1.1481 to 3.4605) was significantly associated with incomplete normalization of blood pressure. CONCLUSIONS: Patients with hypertension and hypothyroidism have increased aortic stiffness. Aortic stiffness is decreased in all patients, whereas hypertension is completely reversible in 50% of patients by hormone replacement therapy. Sustained hypertension may be due to increased aortic stiffness. PMID- 11923810 TI - Blood pressure control and hormone replacement therapy in postmenopausal women at risk for coronary heart disease. AB - BACKGROUND: Coronary heart disease (CHD) in women is strongly associated with estrogen deprivation. For example, risk for CHD increases dramatically after menopause. However, the role of hormone replacement therapy (HRT) in CHD prevention currently is unresolved. To better understand CHD in women, the precise mechanisms by which estrogen affects circulatory function require clarification. Evidence suggests that exogenous estrogen may affect blood pressure (BP) control, but its interaction with other CHD risk factors has not been systematically characterized. The present study examines the role of mildly elevated resting BP, family history of CHD, and HRT on BP responses to stress in postmenopausal women. METHODS: Postmenopausal women on long-term HRT were recruited along with a control group of postmenopausal women not on HRT. These women were divided into higher versus lower risk for CHD on the basis of resting BP and family history of CHD. BP control mechanisms were assessed before, during, and after a computer-controlled laboratory stressor. RESULTS: Results indicate that women with elevated resting BP and positive family history of CHD have exaggerated BP reactivity to stress and that HRT inhibits this effect. CONCLUSIONS: This study suggests that unmedicated postmenopausal women with mildly elevated resting BP and positive family history of CHD have altered BP control as indicated by exaggerated BP responses to stress. HRT eliminates the cumulative effect of resting BP and family history on BP reactivity, suggesting that the circulatory effects of estrogen replacement may operate, at least in part, through normalization of BP reactivity in higher-risk postmenopausal women. PMID- 11923812 TI - Comparative efficacy of fibrinogen and platelet supplementation on the in vitro reversibility of competitive glycoprotein IIb/IIIa receptor-directed platelet inhibition. AB - BACKGROUND: Platelet surface glycoprotein IIb/IIIa (alphaIIb/beta3) receptor inhibition, with prevention of fibrinogen binding and platelet aggregation, concomitantly attenuates arterial thrombotic capacity and impairs protective hemostasis, 2 divergent platelet-dependent processes. PURPOSE: Because the currently available, Food and Drug Administration-approved small molecule glycoprotein IIb/IIIa receptor antagonists are considered "competitive" inhibitors and there is limited information on the reversibility of platelet inhibition with fibrinogen or platelet supplementation, the following series of in vitro experiments were performed. METHODS AND RESULTS: Washed platelets from 24 healthy volunteers were suspended in tyrodes buffer and incubated with achievable (in vivo) concentrations of either tirofiban or eptifibatide before activation with thrombin receptor agonist peptide (15 micromol/L). Platelet aggregation was inhibited by 40% to 50%, but reversal was achieved with fibrinogen supplementation in a concentration-dependent manner. In a separate series of in vitro experiments, platelet inhibition exceeding 90% was established with tirofiban (average concentration 9.28 microg/L) and eptifibatide (average concentration 95.4 microg/L). Recovery of platelet aggregation to at least 50% was achieved after the addition of fibrinogen (0.76-0.80 g/L), platelets (2.4 x 10(11)/L), or their combination. There was an inverse relationship between plasma baseline fibrinogen and the amount of supplemental fibrinogen needed to restore platelet aggregability (r = -0.60; P <.01). CONCLUSION: The reversibility of glycoprotein IIb/IIIa-directed platelet inhibition is influenced by cell surface receptor availability and intrinsic pharmacodynamic mechanism of action. Fibrinogen supplementation with fresh frozen plasma or cryoprecipitate either alone or in combination with platelet transfusion represents an important and readily available treatment consideration for restoring hemostatic potential and managing major hemorrhagic complications associated with the administration of small-molecule, competitive glycoprotein IIb/IIIa receptor antagonists. PMID- 11923813 TI - Increased postprandial lipemia in patients with normolipemic peripheral arterial disease. AB - METHODS: We evaluated the postprandial lipid metabolism in patients with normolipemic peripheral arterial disease (PAD) after the administration of an oral fat load. Total cholesterol, low-density lipoprotein cholesterol (LDL-C), high-density lipoprotein cholesterol (HDL-C), HDL2 and HDL3 subfractions, triglycerides (TGs), lipoprotein(a), and LDL size were determined at baseline and for 8 hours after the meal. RESULTS: In patients with PAD, TGs increased significantly at the 4th, 6th, and 8th hours postprandially; in control subjects, TGs increased at the 4th and 6th hours. HDL decreased significantly at the 4th, 6th, and 8th hours in patients with PAD and at the 6th hour in control subjects. The magnitude of postprandial lipemia, expressed as "the area under the incremental curve for TGs," was higher in patients with PAD than in control subjects (770 +/- 476 vs 391 +/- 195 mg/dL at 8 hours, P <.05). Multiple regression analysis showed that baseline TGs were positively related to the magnitude of postprandial lipemia (P =.01) and that LDL size was negatively related (P =.05). CONCLUSIONS: This is the first documentation of postprandial behavior in patients with normolipemic PAD, suggesting the relevance of postprandial lipoprotein metabolism in the pathogenesis of peripheral atherosclerosis. PMID- 11923814 TI - Poor prognosis and related factors in adults with Eisenmenger syndrome. AB - BACKGROUND: Little information is available regarding the clinical history and long-term prognosis of patients with Eisenmenger syndrome without diagnosis until adulthood. The purpose of this study was to investigate the long-term prognosis and factors predictive of the prognosis in patients with Eisenmenger syndrome diagnosis in adulthood. METHODS AND RESULTS: We retrospectively reviewed the clinical data of 106 patients with Eisenmenger syndrome diagnosis in adulthood. Presence of congenital heart defects had neither been noticed nor confirmed with cardiac catheterization until adulthood in most of the patients in our study. After the diagnosis was confirmed, 42 patients died during a mean follow-up period of 7.6 years. The mean survival time from diagnostic catheterization to death was 5.4 years in nonsurvivors. Survival rate was 98% at 1 year, 77% at 5 years, and 58% at 10 years, with the Kaplan-Meier method. Elevated right atrial pressure (>7 mm Hg) and reduced systemic blood flow (<2.9 L/min) were independently associated with increased mortality rate of adults with Eisenmenger syndrome. CONCLUSION: Patients with Eisenmenger syndrome without diagnosis until adulthood had a poor prognosis. Reduced systemic blood flow and elevated right atrial pressure were associated with high mortality rates in adults with Eisenmenger syndrome. PMID- 11923815 TI - Morphologic changes in the microcirculation induced by chronic smoking habit: a videocapillaroscopic study on the human labial mucosa. AB - BACKGROUND: Cigarette smoking has been identified as a major risk factor for cardiovascular and pulmonary diseases. Although an impressive literature does exist on the subject, no evidence is available on morphologic changes induced with chronic smoking habit in the human microcirculation. SUBJECTS AND METHODS: One hundred healthy subjects, 50 smokers and 50 nonsmokers, underwent videocapillaroscopy of the labial mucosa. For each subject, the caliber of capillary loops, the number of visible capillary loops, the background optical transmittance, the tortuousness of capillary loops, the presence of microaneurysms, the presence of microhemorrhages, and the cumulative smoking habit (pack-year index) were noted. RESULTS: Smokers had a lower caliber of capillary loops (P <.001), with a higher number of visible capillary loops (P <.001), a lower background optical transmittance (P <.001), and a more marked tortuousness of capillary loops (P <.001). Microaneurysms and microhemorrhages were absent in nonsmokers, and 1 smoker of 3 had microaneurysms alone, and 1 smoker of 3 had both microaneurysms and microhemorrhages. A significant correlation was found between cumulative smoking habit and tortuousness of capillary loops (P <.001) and between cumulative smoking habit and total score (tortuousness score + microaneurysm score + microhemorrhage score; P <.005). CONCLUSION: Chronic smoking habit does induce significant morphologic changes in the microcirculation of the human labial mucosa, and these changes can be easily and comfortably recorded with videocapillaroscopy. PMID- 11923817 TI - The numerology of idiocy. PMID- 11923818 TI - Trivial pursuits. PMID- 11923816 TI - Low molecular weight heparin (bemiparin sodium) and the coagulation profile of patients with heart failure. AB - BACKGROUND: Congestive heart failure (CHF) is associated with a hypercoagulable state. PATIENTS AND METHODS: A single-center, randomized, double-blind, placebo controlled trial was performed to test the hypothesis that a prophylactic dose of low molecular weight heparin (bemiparin sodium 3500 IU/daily subcutaneously) will modify a hypercoagulable state in CHF. This study included 100 patients with CHF (New York Heart Association classification II to IV). All patients underwent 3 blood tests, at baseline (before randomization), 24 hours after randomization, and before hospital discharge or within 10 days from randomization. RESULTS: In comparison of baseline bemiparin sodium 3500 IU/daily subcutaneously with after 24 hours, there was a significant decrease in plasma levels of D-dimer (-13.8 ng/mL; P =.01) and prothrombin fragments 1 and 2 (-0.11 nmol/L; P =.01), whereas protein C was significantly increased (+3.5%; P =.03). In comparison of baseline bemiparin sodium 3500 IU/daily subcutaneously with after 4 to 10 days of therapy, there were significantly decreased plasma levels for factor VII:c (-3.0%; P =.01), D-dimer (-44.0 ng/mL; P =.002), and thrombin-antithrombin complex (-0.7 microg/L; P =.0001), whereas protein C was significantly increased (+16.0%; P =.03). On the other hand, in the group of patients treated with placebo after 24 hours, a significant decrease was observed of protein C (-4.0%; P =.04). After 24 hours, the changes from baseline were significantly different for some of the hemostatic factors in comparison of bemiparin sodium 3500 IU/daily and placebo (factor VII:c: -1.7 versus 0.0%; P =.04; D-dimer: -14 versus +24.3 ng/mL; P =.009; prothrombin fragments 1 and 2: -0.11 versus +0.11 nmol/L; P =.01; protein C: +3.5 versus -4.0%; P =.01). Also at discharge, the changes from baseline were different for some of the markers in comparison of bemiparin sodium with placebo (D-dimer: -44 versus 3.8 ng/mL; P =.002; thrombin-antithrombin complex: -0.70 versus +0.14 microg/L; P =.002; protein C: +16.0 versus +0.5%; P =.02). CONCLUSION: Our findings suggest that a hypercoagulable state in heart failure can be modified with bemiparin sodium therapy. PMID- 11923819 TI - US wary of South Korean plans for Californian biocenter. PMID- 11923820 TI - US biotech prepares to fight generic biologics. PMID- 11923821 TI - Nutraceuticals tread business tightrope in Europe. PMID- 11923822 TI - National academy panel urges USDA to toughen reviews of transgenic plants. PMID- 11923823 TI - European Union in disarray over GM seeds. PMID- 11923824 TI - Japanese rules hinder commercialization of publicly funded projects. PMID- 11923826 TI - Prescription for an ailing pharmaceutical industry. PMID- 11923827 TI - Protecting privacy while revealing data. PMID- 11923828 TI - April Fools--sell-side analysts discover biotechnology. PMID- 11923830 TI - Standards for modeling. PMID- 11923831 TI - Wheat, rye, and barley on the cob? PMID- 11923834 TI - Cell-based therapies go live. PMID- 11923835 TI - Trading "wet-work" for network. PMID- 11923836 TI - Alternative splicing in parallel. PMID- 11923837 TI - Turning back the developmental clock. PMID- 11923838 TI - DNA sandwiches with silver and gold. PMID- 11923840 TI - Profiling alternative splicing on fiber-optic arrays. AB - The human transcriptome is marked by extensive alternative mRNA splicing and the expression of many closely related genes, which may be difficult to distinguish using standard microarray techniques. Here we describe a sensitive and specific assay for parallel analysis of mRNA isoforms on a fiber-optic microarray platform. The method permits analysis of mRNA transcripts without prior RNA purification or cDNA synthesis. Using an endogenously expressed viral transcript as a model, we demonstrated that the assay readily detects mRNA isoforms from as little as 10-100 pg of total cellular RNA or directly from a few cells. Multiplexed analysis of human cancer cell lines revealed differences in mRNA splicing and suggested a potential autocrine mechanism in the development of choriocarcinomas. Our approach may be useful in the large-scale analysis of the role of alternative splicing in development and disease. PMID- 11923842 TI - Cloned rabbits produced by nuclear transfer from adult somatic cells. AB - We have developed a method to produce live somatic clones in the rabbit, one of the mammalian species considered up to now as difficult to clone. To do so, we have modified current cloning protocols proven successful in other species by taking into account both the rapid kinetics of the cell cycle of rabbit embryos and the narrow window of time for their implantation after transfer into foster recipients. Although our method still has a low level of efficiency, it has produced several clones now proven to be fertile. Our work indicates that cloning can probably be carried out successfully in any mammalian species by taking into account physiological features of their oocytes and embryos. Our results will contribute to extending the use of rabbit models for biomedical research. PMID- 11923841 TI - Multiplexed protein profiling on microarrays by rolling-circle amplification. AB - Fluorescent-sandwich immunoassays on microarrays hold appeal for proteomics studies, because equipment and antibodies are readily available, and assays are simple, scalable, and reproducible. The achievement of adequate sensitivity and specificity, however, requires a general method of immunoassay amplification. We describe coupling of isothermal rolling-circle amplification (RCA) to universal antibodies for this purpose. A total of 75 cytokines were measured simultaneously on glass arrays with signal amplification by RCA with high specificity, femtomolar sensitivity, 3 log quantitative range, and economy of sample consumption. A 51-feature RCA cytokine glass array was used to measure secretion from human dendritic cells (DCs) induced by lipopolysaccharide (LPS) or tumor necrosis factor-alpha (TNF-alpha). As expected, LPS induced rapid secretion of inflammatory cytokines such as macrophage inflammatory protein (MIP)-1beta, interleukin (IL)-8, and interferon-inducible protein (IP)-10. We found that eotaxin-2 and I-309 were induced by LPS; in addition, macrophage-derived chemokine (MDC), thymus and activation-regulated chemokine (TARC), soluble interleukin 6 receptor (sIL-6R), and soluble tumor necrosis factor receptor I (sTNF-RI) were induced by TNF-alpha treatment. Because microarrays can accommodate approximately 1,000 sandwich immunoassays of this type, a relatively small number of RCA microarrays seem to offer a tractable approach for proteomic surveys. PMID- 11923843 TI - Computational modeling of the dynamics of the MAP kinase cascade activated by surface and internalized EGF receptors. AB - We present a computational model that offers an integrated quantitative, dynamic, and topological representation of intracellular signal networks, based on known components of epidermal growth factor (EGF) receptor signal pathways. The model provides insight into signal-response relationships between the binding of EGF to its receptor at the cell surface and the activation of downstream proteins in the signaling cascade. It shows that EGF-induced responses are remarkably stable over a 100-fold range of ligand concentration and that the critical parameter in determining signal efficacy is the initial velocity of receptor activation. The predictions of the model agree well with experimental analysis of the effect of EGF on two downstream responses, phosphorylation of ERK-1/2 and expression of the target gene, c-fos. PMID- 11923844 TI - Identification of genes that function in the TNF-alpha-mediated apoptotic pathway using randomized hybrid ribozyme libraries. AB - Now that the sequences of many genomes are available, methods are required for the rapid identification of functional genes. We describe here a simple system for the isolation of genes that function in the tumor necrosis factor-alpha (TNF alpha)-mediated pathway of apoptosis, using RNA helicase-associated ribozyme libraries with randomized substrate-binding arms. Because target-site accessibility considerably limits the effective use of intracellular ribozymes, the effectiveness of a conventional ribozyme library has been low. To overcome this obstacle, we attached to ribozymes an RNA motif (poly(A)-tail) able to interact with endogenous RNA helicase(s) so that the resulting helicase-attached, hybrid ribozymes can more easily attack target sites regardless of their secondary or tertiary structures. When the phenotype of cells changes upon introduction of a ribozyme library, genes responsible for these changes may be identified by sequencing the active ribozyme clones. In the case of TNF-alpha mediated apoptosis, when a ribozyme library was introduced into MCF-7 cells, surviving clones were completely or partially resistant to TNF-alpha-induced apoptosis. We identified many pro-apoptotic genes and partial sequences of previously uncharacterized genes using this method. Our gene discovery system should be generally applicable to the identification of functional genes in various systems. PMID- 11923846 TI - Combined transcriptome and genome analysis of single micrometastatic cells. AB - In human cancer, early systemic spread of tumor cells is recognized as a leading cause of death. Adjuvant therapies are administered to patients after complete resectioning of their primary tumors to eradicate the few residual and latent metastatic cells. These therapeutic regimens, however, are currently designed without direct information about the presence or nature of the latent cells. To address this problem, we developed a PCR-based technique to analyze the transcriptome of individual tumor cells isolated from the bone marrow of cancer patients. From the same cells, genomic aberrations were identified by comparative genomic hybridization. The utility of this approach for understanding the biology of occult disseminated cells and for the identification of new therapeutic targets is demonstrated here by the detection of frequent extracellular matrix metalloproteinase inducer (EMMPRIN; CD147) expression which was verified by immunostaining. PMID- 11923845 TI - Engineering N-glycosylation mutations in IL-12 enhances sustained cytotoxic T lymphocyte responses for DNA immunization. AB - Interleukin-12 (IL-12), consisting of p40 and p35 subunits, produces both p70 heterodimer and free p40. p70 is essential for the induction of T-helper 1 (Th1) and cytotoxic T-cell (CTL) immunity, whereas p40 inhibits p70-mediated function. Here, we found that mutations introduced into N-glycosylation sites (N220 of murine p40 and N222 of human p40) reduced secretion of p40 but not p70. Co immunization of N220 mutant mIL-12 gene with hepatitis C virus (HCV) E2 DNA significantly enhanced long-term E2-specific CD8+ T-cell response and protection against tumor challenge compared with that of wild type. Our results indicate that the ratio of p70 to p40 is important for generating sustained long-term cell mediated immunity. Thus, the mutant IL-12 could be utilized for the development of DNA vaccines as an adjuvant for the generation of long-term memory T-cell responses. PMID- 11923847 TI - Genome-wide detection of chromosomal imbalances in tumors using BAC microarrays. AB - Chromosomal imbalances such as deletions and amplifications are common rearrangements in most tumors. Specific rearrangements are consistently associated with specific tumor types or stages, implicating the role of the genes in a region of chromosomal imbalance in tumor initiation and progression. The development of comparative genomic hybridization (CGH) has obviated the need to obtain metaphase spreads from tumors, so that the chromosomal imbalances in many solid tumors may be revealed using an extracted genomic DNA sample. However, the resolution of the cytogenetic method remains and the extreme technical difficulty of CGH has restricted its use. Conceptually, DNA microarray-based CGH is an obvious solution to all of the limitations of conventional CGH. Although arrays have been used for CGH studies, their success has been limited by poor specific signal-to-noise ratios. Here we demonstrate a microarray-based CGH method that allows reliable detection of chromosomal deletions and amplifications with high resolution. Our microarray system is fundamentally different from most current microarray technologies in that activated DNA is printed on natural glass surfaces while other systems almost exclusively focus on activating the surfaces, a strategy that invariably introduces hybridization backgrounds. The concept of using pre-modification may be generally applied for making arrays of other biological materials, as modifying the substrates will be more controllable in solution than on surfaces. PMID- 11923849 TI - Nontraditional publications and their effect on patentable inventions. PMID- 11923848 TI - Expression of exogenous protein in the egg white of transgenic chickens. AB - Using a replication-deficient retroviral vector based on the avian leukosis virus (ALV), we inserted into the chicken genome a transgene encoding a secreted protein, beta-lactamase, under the control of the ubiquitous cytomegalovirus (CMV) promoter. Biologically active beta-lactamase was secreted into the serum and egg white of four generations of transgenic chickens. The expression levels were similar in successive generations, and expression levels in the magnum of the oviduct were constant over at least 16 months in transgenic hens, indicating that the transgene was stable and not subject to silencing. These results support the potential of the hen as a bioreactor for the production of commercially valuable, biologically active proteins in egg white. PMID- 11923853 TI - Careers in regulatory affairs: from practitioner to professional. PMID- 11923855 TI - FDA requests comments on extralabel drug use in animals. PMID- 11923856 TI - For want of some references... PMID- 11923857 TI - Mass in the oral cavity of cultured Summer flounder. PMID- 11923859 TI - A virtual tour of the Guide for zebrafish users. AB - PHS-funded and AAALAC-accredited facilities are required to use the Guide as the basis for setting up a zebrafish care and use program. The authors describe how they accomplished this task at the University of Oregon Zebrafish Facility. PMID- 11923860 TI - Diagnostic techniques for clinical investigation of laboratory zebrafish. AB - Unexpected morbidity and mortality of aquatic animal models represent a significant problem for researchers. The authors outline the basic procedures used to diagnose disease outbreaks in laboratory zebrafish colonies, and provide a basic framework for initiating clinical investigations. PMID- 11923861 TI - Use of platyfishes and swordtails in biological research. AB - The use of platyfishes and swordtails as research models is on the rise. The authors review the basic biology and care of these animals and describe their common uses in research. PMID- 11923862 TI - Recirculating systems for zebrafish. PMID- 11923864 TI - FDA publishes validation guide for electronic records and signatures. PMID- 11923863 TI - Peering into the brain. PMID- 11923865 TI - The letter is in the mail, right? PMID- 11923866 TI - Dyspnea in Yorkshire-cross weanling swine. Diagnosis: ventricular septal defect. PMID- 11923867 TI - Samples, sample selection, and statistics: living with uncertainty. AB - The author explains how to determine how many animals in a population need to be sampled when monitoring for pathogens. He concludes that even with perfect testing programs, there is always a chance that an infected animal will be missed. PMID- 11923868 TI - Diagnostic molecular microbiology in laboratory animal health monitoring and surveillance programs. AB - Molecular microbiology provides a rapid, reliable, sensitive, and specific means of detecting pathogens in laboratory animals. The author discusses the interpretation of the results of molecular microbiological testing done in conjunction with serologic, bacteriologic, and histopathologic tests, focusing on molecular microbiology as it fits in an overall program of animal health diagnostic profiling. PMID- 11923869 TI - Damage control: a guide to dealing with an infectious break. AB - A change in the microbial status of laboratory animals can represent a disruptive event in the research process. The author suggests a sequence of events from the time a facility learns of a potential infectious "break," through investigation of its source, and its ultimate control. PMID- 11923870 TI - Modification of the anal tape method for detection of pinworms in rodents. AB - A modification of Graham's anal tape technique for detection of oxyurid worm eggs involves a coverslip or microscope slide covered with a thin layer of adhesive. The worm eggs stick to the adhesive and can be directly observed under the microscope. PMID- 11923871 TI - Severe impairment of interleukin-1 and Toll-like receptor signalling in mice lacking IRAK-4. AB - Toll-like receptors (TLRs), which recognize pathogen-associated molecular patterns, and members of the pro-inflammatory interleukin-1 receptor (IL-1R) family, share homologies in their cytoplasmic domains called Toll/IL-1R/plant R gene homology (TIR) domains. Intracellular signalling mechanisms mediated by TIRs are similar, with MyD88 (refs 5-8) and TRAF6 (refs 9, 10) having critical roles. Signal transduction between MyD88 and TRAF6 is known to involve the serine threonine kinase IL-1 receptor-associated kinase 1 (IRAK-1) and two homologous proteins, IRAK-2 (ref. 12) and IRAK-M. However, the physiological functions of the IRAK molecules remain unclear, and gene-targeting studies have shown that IRAK-1 is only partially required for IL-1R and TLR signalling. Here we show by gene-targeting that IRAK-4, an IRAK molecule closely related to the Drosophila Pelle protein, is indispensable for the responses of animals and cultured cells to IL-1 and ligands that stimulate various TLRs. IRAK-4-deficient animals are completely resistant to a lethal dose of lipopolysaccharide (LPS). In addition, animals lacking IRAK-4 are severely impaired in their responses to viral and bacterial challenges. Our results indicate that IRAK-4 has an essential role in innate immunity. PMID- 11923872 TI - Deubiquitination of p53 by HAUSP is an important pathway for p53 stabilization. AB - The p53 tumour suppressor is a short-lived protein that is maintained at low levels in normal cells by Mdm2-mediated ubiquitination and subsequent proteolysis. Stabilization of p53 is crucial for its tumour suppressor function. However, the precise mechanism by which ubiquitinated p53 levels are regulated in vivo is not completely understood. By mass spectrometry of affinity-purified p53 associated factors, we have identified herpesvirus-associated ubiquitin-specific protease (HAUSP) as a novel p53-interacting protein. HAUSP strongly stabilizes p53 even in the presence of excess Mdm2, and also induces p53-dependent cell growth repression and apoptosis. Significantly, HAUSP has an intrinsic enzymatic activity that specifically deubiquitinates p53 both in vitro and in vivo. In contrast, expression of a catalytically inactive point mutant of HAUSP in cells increases the levels of p53 ubiquitination and destabilizes p53. These findings reveal an important mechanism by which p53 can be stabilized by direct deubiquitination and also imply that HAUSP might function as a tumour suppressor in vivo through the stabilization of p53. PMID- 11923873 TI - Bop encodes a muscle-restricted protein containing MYND and SET domains and is essential for cardiac differentiation and morphogenesis. AB - Many transcription factors regulate specific temporal-spatial events during cardiac differentiation; however, the mechanisms that regulate such events are largely unknown. Using a modified subtractive hybridization method to identify specific genes that influence early cardiac development, we found that Bop is expressed specifically in cardiac and skeletal muscle precursors before differentiation of these lineages. Bop encodes a protein containing MYND and SET domains, which have been shown to regulate transcription by mediating distinct chromatin modifications. We show that m-Bop is a histone deacetylase-dependent transcriptional repressor. Targeted deletion of Bop in mice disrupted maturation of ventricular cardiomyocytes and interfered with formation of the right ventricle. Normal expression of Hand2, a transcription factor essential for right ventricular development, in cardiomyocyte precursors is dependent upon m-Bop. These results indicate that m-Bop is essential for cardiomyocyte differentiation and cardiac morphogenesis. PMID- 11923874 TI - Defective prelamin A processing and muscular and adipocyte alterations in Zmpste24 metalloproteinase-deficient mice. AB - The mouse ortholog of human FACE-1, Zmpste24, is a multispanning membrane protein widely distributed in mammalian tissues and structurally related to Afc1p/ste24p, a yeast metalloproteinase involved in the maturation of fungal pheromones. Disruption of the gene Zmpste24 caused severe growth retardation and premature death in homozygous-null mice. Histopathological analysis of the mutant mice revealed several abnormalities, including dilated cardiomyopathy, muscular dystrophy and lipodystrophy. These alterations are similar to those developed by mice deficient in A-type lamin, a major component of the nuclear lamina, and phenocopy most defects observed in humans with diverse congenital laminopathies. In agreement with this finding, Zmpste24-null mice are defective in the proteolytic processing of prelamin A. This deficiency in prelamin A maturation leads to the generation of abnormalities in nuclear architecture that probably underlie the many phenotypes observed in both mice and humans with mutations in the lamin A gene. These results indicate that prelamin A is a specific substrate for Zmpste24 and demonstrate the usefulness of genetic approaches for identifying the in vivo substrates of proteolytic enzymes. PMID- 11923875 TI - beta-cell-specific deletion of the Igf1 receptor leads to hyperinsulinemia and glucose intolerance but does not alter beta-cell mass. AB - Regulation of glucose homeostasis by insulin depends on the maintenance of normal beta-cell mass and function. Insulin-like growth factor 1 (Igf1) has been implicated in islet development and differentiated function, but the factors controlling this process are poorly understood. Pancreatic islets produce Igf1 and Igf2, which bind to specific receptors on beta-cells. Igf1 has been shown to influence beta-cell apoptosis, and both Igf1 and Igf2 increase islet growth; Igf2 does so in a manner additive with fibroblast growth factor 2 (ref. 10). When mice deficient for the Igf1 receptor (Igf1r(+/-)) are bred with mice lacking insulin receptor substrate 2 (Irs2(-/-)), the resulting compound knockout mice show a reduction in mass of beta-cells similar to that observed in pancreas of Igf1r(-/ ) mice (ref. 11), suggesting a role for Igf1r in growth of beta-cells. It is possible, however, that the effects in these mice occur secondary to changes in vascular endothelium or in the pancreatic ductal cells, or because of a decrease in the effects of other hormones implicated in islet growth. To directly define the role of Igf1, we have created a mouse with a beta-cell-specific knockout of Igf1r (betaIgf1r(-/-)). These mice show normal growth and development of beta cells, but have reduced expression of Slc2a2 (also known as Glut2) and Gck (encoding glucokinase) in beta-cells, which results in defective glucose stimulated insulin secretion and impaired glucose tolerance. Thus, Igf1r is not crucial for islet beta-cell development, but participates in control of differentiated function. PMID- 11923878 TI - Impact of environmental endocrine-active substances on public health. PMID- 11923879 TI - Genetic polymorphisms and metabolism of endocrine disruptors in cancer susceptibility. AB - Epidemiological studies have estimated that approximately 80% of all cancers are related to environmental factors. Individual cancer susceptibility can be the result of several host factors, including differences in metabolism, DNA repair, altered expression of tumor suppressor genes and proto-oncogenes, and nutritional status. Xenobiotic metabolism is the principal mechanism for maintaining homeostasis during the body's exposure to xenobiotics. The balance of xenobiotic absorption and elimination rates in metabolism can be important in the prevention of DNA damage by chemical carcinogens. Thus the ability to metabolize and eliminate xenobiotics can be considered one of the body's first protective mechanisms. Variability in individual metabolism has been related to the enzymatic polymorphisms involved in activation and detoxification of chemical carcinogens. This paper is a contemporary literature review on genetic polymorphisms involved in the metabolism of endocrine disruptors potentially related to cancer development. PMID- 11923880 TI - On the rumors about the silent spring. Review of the scientific evidence linking occupational and environmental pesticide exposure to endocrine disruption health effects. AB - Occupational exposure to some pesticides, and particularly DBCP and chlordecone, may adversely affect male fertility. However, apart from the therapeutic use of diethylstilbestrol, the threat to human reproduction posed by "endocrine disrupting" environmental contaminants has not been supported by epidemiological evidence thus far. As it concerns other endocrine effects described in experimental animals, only thyroid inhibition following occupational exposure to amitrole and mancozeb has been confirmed in humans. Cancer of the breast, endometrium, ovary, prostate, testis, and thyroid are hormone-dependent, which fostered research on the potential risk associated with occupational and environmental exposure to the so-called endocrine-disrupting pesticides. The most recent studies have ruled out the hypothesis of DDT derivatives as responsible for excess risks of cancer of the reproductive organs. Still, we cannot exclude a role for high level exposure to o,p'-DDE, particularly in post-menopausal ER+ breast cancer. On the other hand, other organochlorine pesticides and triazine herbicides require further investigation for a possible etiologic role in some hormone-dependent cancers. PMID- 11923881 TI - Secular variations in sperm quality: fact or science fiction? AB - The debate concerning the possible degradation in human sperm quality began in the 1970s, was revived at the beginning of the 1990s and has continued to mobilize the scientific community ever since. After the meta-analysis by Carlsen et al. (1992) showing a decline in human semen quality over the last 50 years, several groups investigated the sperm characteristics of more or less homogeneous groups of men who had provided semen at the same center for 10 to 20 years. A significant decrease in sperm concentration was reported in some studies, but not in others. Meanwhile, there is an increasing number of reports suggesting that physical and chemical factors introduced and spread by human activity in the environment may have contributed to sperm decline. At the end of the 20th century the debate on declining semen quality is not closed. The lack of certainty and the serious consequences that such a decline would have on the fertility of human populations make this an important public health issue at the start of the 21st century. For this reason, intensive research should be developed in both fundamental and epidemiological domains, particularly in South America, where industrial and agricultural pollution pose a serious threat to the population. PMID- 11923882 TI - Changes in male reproductive health and effects of endocrine disruptors in Scandinavian countries. AB - Male reproductive health has deteriorated in many ways during the last decades. The incidence of testicular cancer has rapidly increased in Europe and European derived populations. Sperm concentrations have declined and sperm motility and morphology have worsened in many areas. Both adverse trends have been shown to be associated with year of birth. Older birth cohorts have better reproductive health than the younger generations. Incidences of cryptorchidism and hypospadias have also increased according to several studies. The reasons for secular trends are unknown, but the rapid pace of the change points to environmental causes. Endocrine disrupting chemicals have been hypothesized to influence male reproductive health. PMID- 11923883 TI - Endocrine disruptors and congenital anomalies. AB - The specialized literature was reviewed concerning the suspected increasing secular trends in the frequency of female births, male genital congenital anomalies, abnormal sperm counts, and testicular cancer. Although no risk factors could be identified yet, the observed sex ratio decline during the last decades has been considered to be an effect of certain pollutants on normal hormone activity, and human reproductive development. Reported increasing trends in the frequencies of hypospadias and cryptorchidism are very difficult to be interpreted due to the large variability in the registered frequency of these malformations due to operational as well as biological reasons. PMID- 11923884 TI - Rationale for the study of the human sex ratio in population studies of polluted environments. AB - The human secondary sex ratio remains a subject of substantial interest. The possibility has been raised that environmental chemical exposures have played a role in the changes associated with the sex ratio in a number of countries. The possibility that such an effect may be present is supported at least theoretically by the observation that clomiphene citrate, a drug used in the treatment of infertility with powerful estrogenic and anti-estrogenic properties, has profound effects on the sex ratio resulting in significantly fewer males at birth. Using a model of causality based on the clinical identification of adverse drug effect methodology one may improve the objectivity of the assessment of significant environmental exposures on this human reproductive outcome. PMID- 11923886 TI - Food consumption and adipose tissue DDT levels in Mexican women. AB - This article analyzes food consumption in relation to levels of DDE (the principal metabolite of DDT) in the adipose tissue of 207 Mexican women residing in States with high and low exposure to DDT. Data on the women's dietary habits and childbearing history were obtained from a personal interview. Adipose tissue DDE levels were measured by gas-liquid chromatography and compared by analysis of variance (ANOVA) and multiple linear regression. Adipose tissue DDE levels increased significantly with age (p = 0.005) and residence in coastal areas (p = 0.002) and non-significantly with the consumption of onion, cauliflower, prickly pear, squash blossoms, sweet corn, broad beans, chili pepper sauce, ham, and fish. Even so, during breastfeeding there was a non-significant reduction in these levels. The findings suggest that certain foods serve as vehicles for DDE residues and confirm that breastfeeding is a mechanism for the elimination of this insecticide, which accumulates over the years in the human body. PMID- 11923885 TI - Human reproductive system disturbances and pesticide exposure in Brazil. AB - The observation of reproductive disturbances in humans and in the wildlife has been reported in the last decade in different countries. Exposure to different chemicals possibly acting in the endocrine system or endocrine disruptors, including pesticides, has been a hypothesis raised to explain the observed changes. This paper aimed to present results of an epidemiological ecologic study carried out to explore population data on pesticides exposure in selected Brazilian states in the eighties and human reproductive outcomes in the nineties. Pearson correlation coefficients were ascertained between available data pesticides sales in eleven states in Brazil in 1985 and selected further reproductive outcomes or their surrogates. Moderate to high correlations were observed to infertility, testis, breast, prostate and ovarian cancer mortality. Despite the restrains of ecologic studies to establish cause-effect relationships, the observed results are in agreement with evidence supporting a possible association between pesticides exposure and the analyzed reproductive outcomes. PMID- 11923887 TI - Dioxin exposure and porcine reproductive hormonal activity. AB - To characterize the action of 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) during both the follicular and luteal phases of the ovarian cycle, the direct effect of TCDD was investigated in vitro using a system of primary monolayer cell culture. Granulosa and theca cells were collected from the preovulatory follicles and cultured as a co-culture, thus resembling follicles in vivo. Luteal cells were isolated from the corpora lutea collected during the midluteal phase. In both cases cells were isolated from the ovaries of animals exhibiting natural estrus cycle. Results of these experiments suggest that TCDD decreases estradiol secretion by follicular cells and progesterone secretion by luteal cells in a dose-dependent manner. It was also shown that TCDD disrupts steroidogenesis through its influence on the activity of enzymes involved in the steroid biosynthesis cascade. In luteal cells, its action is mediated via the aryl hydrocarbon receptor (AhR) and is probably independent of estrogen receptor (ER) stimulation. Endocrine disruptors that interfere with estradiol production in the follicles can act as ovulatory disruptors, and while interfering with progesterone production by luteal cells they can act as abortifacients. PMID- 11923888 TI - Occurrence of imposex in Thais haemastoma: possible evidence of environmental contamination derived from organotin compounds in Rio de Janeiro and Fortaleza, Brazil. AB - There are indications that the widespread use of organotin compounds (TBT and TPT) as antifoulings, as stabilizers in plastic and as pesticides, has severely affected several species of marine organisms. The most striking effect of TBT and TPT as hormonal disruptors is the development of male organs in females of gastropods, currently denominated imposex. This syndrome can lead to the sterilization and death of affected organisms. The present work gives an overview of the present state of knowledge on imposex occurrence and reports results of a survey conducted in Guanabara Bay, Rio de Janeiro and in several sites along the coast of Fortaleza, Ceara State. Different stages of imposex development were verified in this survey, however, the most prominent levels appeared associated to known spot sources of TBT and TPT. PMID- 11923889 TI - Organic micropollutants on river sediments from Rio de Janeiro State, Southeast Brazil. AB - The paper is a contribution for the knowledge upon concentrations and fate of different kinds of organic micropollutants in Tropical River system from a very industrialized region in Brazil. The presented data was obtained during three years of an International Research Project between Brazilian and Dutch institutions. The sediments were sampled at the Paraiba do Sul-Guandu river watershed, the most important watercourse of Rio de Janeiro state, where up 90% of the population depends on its water for domestic uses. After extraction with non-polar solvents in a hot sohxlet device and clean up using chromatographic columns, three classes of organic micropollutants were analyzed: organochlorine insecticides (OCs), polychlorinated biphenyls (PCBs) and polynuclear aromatic hydrocarbons (PAHs). The organochlorines, including the PCBs were scarcely present in the collected samples probably reflecting the restrictions of use of this class of compounds in the Brazilian market. However, the PAHs levels were high at the vicinity of a huge steelworks located in the city of Volta Redonda. This contamination is probably due to the massive use of coal in the above-cited metallurgical plant. PMID- 11923890 TI - Endocrine disrupting chemicals: harmful substances and how to test them. AB - This paper presents an analysis of the opinions of different groups from: scientists, international regulatory bodies, non-governmental organizations and industry; with an interest in the problem of identifying chemical substances with endocrine disrupting activity. There is also discussion of the consequences that exposure to endocrine disruptors may have for human health, considering concrete issues related to: the estimation of risk; the tests that must be used to detect endocrine disruption; the difficulties to establish an association between dose, time of exposure, individual susceptibility, and effect; and the attempts to create a census of endocrine disruptors. Finally, it is proposed that not all hormonal mimics should be included under the single generic denomination of endocrine disruptors. PMID- 11923891 TI - Organizational and activational effects of estrogenic endocrine disrupting chemicals. AB - Endocrine disruption is a hypothesis of common mode of action that may define a set of structurally varied chemicals, both natural and synthetic. Their common mode of action may suggest that they produce or contribute to similar toxic effects, although this has been difficult to demonstrate. Insights from developmental biology suggest that development of hormone sensitive systems, such as the brain and the genitourinary tract, may be particularly sensitive to EDCs. Because these systems are both organized and later activated by hormones, the brain and vagina may be valuable model systems to study the toxicity of EDCs in females and to elucidate mechanisms whereby early exposures appear to affect long term function. PMID- 11923892 TI - Endocrine disruptors and dental materials: health implications associated with their use in Brazil. AB - This study analyzes international historical trends in the use of different types of materials in dental practice. The author describes the chemical properties of their ingredients and the potential and observed adverse effects in patients and dental technicians resulting from clinical or occupational exposure to various metals like beryllium, used to produce metal alloys. The growing use of various products (resin cements, ionomer cements, aesthetic restorative materials, resins, endodontal cements, and others) based on the compound bisphenol-A, whose chemical structure is similar to that of estrogen. Considering the demographic and contemporary work force characteristics of those involved in dental practice in the Brazil, the study highlights the possible effect of the use of these materials in both male and female patients and all age strata, as well as in health professionals with occupational exposure to products containing bisphenol A. PMID- 11923893 TI - Health surveillance and endocrine disruptors. AB - The author discusses the extreme relevance of research on the presence of endocrine disruptors (EDs) in products of interest to health surveillance (HS). Focusing on EDs, the author highlights the urgency of changes already under way in the direction of HS. The shift should be from product and product-registration approaches to the productive process and its realization in consumption, generation of contaminants, and alterations in the health of workers and the overall population. He briefly describes: regulatory gaps for dealing with EDs; difficulty in evaluating risk and suspension of the production and use of products with its characteristics and the need, as exemplified by such products, to enhance the inter-relationship among all stakeholders and to turn HS into a state-of-the-art technological setting, associated with the academic community and accountable to the public. The author reports on measures already taken in relation to EDs, including the establishment of a reference laboratory for analyzing persistent organic pollutants (POPs), interruption of the use of various POPs in Brazil and an initial review of requirements for registering pesticides under the Brazilian National Health Surveillance Agency (ANVISA). PMID- 11923895 TI - Sperm characteristics in a sample of healthy adolescents in Sao Paulo, Brazil. AB - The article presents preliminary data from a prospective investigation in a sample of healthy 14-17-year-old students from a technical school in Sao Paulo, Brazil. Ninety-six Tanner stage 5 and thirty-one Tanner stage 4 adolescents were evaluated: testicular volume was measured using a Prader orchidometer, and semen analysis was performed according to standard procedures. Median testicular volume was 20.0 ml among Tanner 5 students and 15.0 ml in Tanner 4 students in both the right and left testes. No significant differences were found in sperm volume or motility. Median concentration was 66.0 million/ml for Tanner 5 and 47.0 million/ml for Tanner 4 subjects. Morphological patterns showed abnormal forms in 81.9% of Tanner 5 and 93.6% of Tanner 4 adolescents. Oligospermia (sperm concentration < 5 million/ml) was observed in 7.3% of Tanner stage 5 and 12.9% of Tanner stage 4 individuals. Azoospermia was observed in 3 students (1.8%), with counts less than 1.0 in 8 students (4.8%). The authors discuss the observed results, analyzing the potential implications arising from biological development and potential environmental exposures. PMID- 11923894 TI - Serum levels of organochlorine pesticides and polychlorinated biphenyls among inhabitants of Greater Metropolitan Rio de Janeiro, Brazil. AB - Levels of persistent organochlorine pesticides (OCPs) and polychlorinated biphenyls (PCBs) were determined in the blood serum of people living and working in the urban area of greater Rio de Janeiro city. Blood samples from 33 volunteers (16 males, 17 females, 19-63 years old) were taken in January 1999. OCP residues (op'DDT, pp'DDT, pp'DDD, pp'DDE, Aldrin, Dieldrin, Endrin, Heptachlor, Heptachlor-epoxide, alpha-, beta- and gamma-Hexachlorocyclo-hexane, Hexachlorobenzene) and PCBs (congeners: 28, 52, 101, 138, 153, 180) were extracted with n-hexane and analyzed by gas chromatography with electron capture detection. Except for pp'DDE (detection limit = 1.4 microg/L) no other OCP residue was found in the samples. No PCB congener (detection limit = 2.0 microg/L) was detected either. pp'DDE was found in 17 out of 33 samples in concentrations that ranged from 1.4 to 8.4 microg/L of serum or, on a fat basis, from 0.200 to 3.452 microg/g of serum lipids. Percentage of positive samples (%) and levels of pp'DDE (range of positive samples) increased from the youngest to the oldest group (< or = 29 yrs: 10%, 0.278 microg/g; 30-39 yrs: 60%, 0.200-0.765 microg/g; > or = 40 yrs: 77%, 0.257-3.452 microg/g). PMID- 11923896 TI - A preliminary psychosocial study of Parkinson's patients after fetal tissue transplantation. AB - A novel and promising approach currently available is the grafting of fetal nigra tissues into the brain of a Parkinson's patient. However, the degree of success for such treatment has mainly been evaluated by histological, radiological and physical assessments, with little or no attention paid to its effects on the patient's psychosocial status. In order to provide a more comprehensive profile, a survey was conducted using questionnaires and interviews to examine the physio psycho-social progress of six Parkinson's patients who underwent fetal tissue grafting in Taiwan. Results indicated that the physiological effects varied among patients. Noticeably an improvement in walking was the most gratifying, but progress in dressing and writing ability seemed disappointing. From the psychological point of view, depression was commonly felt by patients. Other feelings included fear, ambivalence, and change in body image. In addition, all six patients found that their relationship with spouse became worse; they also reduced social interaction to the minimum. From the findings, we also noticed that inadequate knowledge contributed to part of the results. Therefore, some nursing suggestions especially educating the patients and family about the facts and outcome of fetal tissue transplantation are highly recommended. These may help Parkinson's patients adjust better after undergoing this novel treatment. PMID- 11923897 TI - Relationship between attendance at breakfast and school achievement among nursing students. AB - This survey by correlation study with partial qualitative design examined the effect of attendance at breakfast with school achievement in 710 first-year nursing students at a nursing institute in Northern Taiwan. Dietary attendance cards were scanned by computer before each meal. Since the school is located at a geographically isolated suburban area, this program was compulsory for all first year nursing students and was included in the tuition fee. Comparison of attendance at breakfast over the four-month semester (from September 1999 to January 2000) and final semester class ranking showed a lower attending rate of breakfast (attendance at breakfast equal to or under 60% of mean times in a 4 month semester) was significantly associated with lower scores on tests in six individual classes and a class rank; while a higher frequency of breakfast attendance (attendance at breakfast equal to or over 85% of mean times in a 4 month semester) was significantly associated with higher scores on tests in six individual classes and a higher class rank (p <.001). To determine the relationships between frequency of attending breakfast and practicing health promotion behaviors, the Chinese Health Promoting Scale was used. We also found that those who skipped breakfast frequently had more negative health promotion behaviors than those who attended breakfast more regularly (p <.001). These results indicate that school achievement is negatively affected when nursing students omit breakfast. The research outcome supports the importance of breakfast in school achievement. PMID- 11923898 TI - Effects of systematic nursing instruction of mothers on using medication and on health status of asthmatic children. AB - The purpose of this study was to investigate the effects of systematic nursing instruction for mothers of children with asthma on their knowledge of the medication, ability to use inhalation devices, self-efficacy of medication use, and on children's health status. A quasi-experimental study was conducted at the Pediatric Allergy Clinic of a major academically based medical center in Taipei, Taiwan. The sample consisted of 82 mothers and their children recruited by purposive sampling. The intervention group (n = 41) received systematic nursing instruction from the researchers and the control group (n = 41) received regular nursing care from staff nurses. The effects of systematic nursing instruction were evaluated two months later using four questionnaires: the mother's knowledge of medication, inhaling steps, self-efficacy of medication use, and child's health status. The results of this study showed that systematic nursing instruction of mothers of children with asthma significantly improved their knowledge of the medication, ability to use inhalation devices, and self efficacy, as well as decreasing the number of episodes of respiratory disease of children over a two month period. To sum up, these findings demonstrate that systematic nursing instruction of mothers about asthma medication not only benefited mothers directly, but also improved children's health status indirectly. PMID- 11923899 TI - Perception of risk by home health care nurses in Taiwan. AB - A study of cross-sectional design was used to understand the level of risk perceived by home health care nurses in Taiwan and its correlators. A total of 413 self-administrated structured questionnaires were sent by mail. 300 responses were received, with 258 valid ones. The response rate was 72.6%. The 90 occupational hazards fell into six risk factors: "professional", "traffic", "geographical", "clients' condition and interaction among clients and families", "law" and "organization". The highest scores among the six factors were associated with "law", "clients' condition and interaction among clients and families", and "organization". Marital status and home health care training were significant variables related to the level of risk perceived. Respondents who were married or had home health care training had higher level of risk perceived in the six risk factors than those who were not married or without any training. The variable of years of experience in home health care had a significantly negative correlation with both level of risk perceived for the factors "professional" and "clients' condition and interaction among clients and families". There was a significant positive correlation between the number of visits made by respondents and level of risk perceived for the traffic and geographical factors. There was no statistically significant correlation between the demographics of respondents and level of risk perceived in law. The variables educational level, whether received home health care training, and whether obtained training certificate showed no significant difference in six levels of risk perceived by respondents. The study proposed several suggestions for practice, education, policy and future study. PMID- 11923900 TI - Parenting stress and parents' willingness to accept treatment in relation to behavioral problems of children with attention-deficit hyperactive disorder. AB - The purpose of this research was to explore parenting stress and parents' willingness to accept treatment in relation to the behavioral problems of children with Attention-Deficit Hyperactive Disorder (ADHD). A total of 100 subjects, either fathers or mothers of children with ADHD, were recruited by convenience sampling from southern Taiwan. The results of this research were: (1) the standardized score for parents' perception of the behavioral problems of their ADHD children was 67.09. (2) The standardized score for parents perception of parenting stress was 62.00. (3) The parents who were most willing to accept treatment were those in the parent training group. (4) The behavioral problems of ADHD children were significantly related to parenting stress (p .001). (5) ADHD children's behavioral problems and parental self-awareness of psychological and emotional problems were the two variables most influential on parenting stress, and recognition of the pathological cause was the most influential factor in willingness to accept treatment. Through this research we also found that the parents hoped that a parent training group would be established. Therefore, nursing personnel can try to develop parent training groups in order to relieve parents' parenting stress. PMID- 11923901 TI - Promoting physical activity among Taiwanese and American adolescents. AB - The purpose of this paper is to present a comparative analysis of the determinants of physical activity among adolescents in Taiwan and in the United States. Pender's Health Promotion Model (HPM) served as the theoretical framework for both studies (Pender, 1996). The major determinants of health behavior in the HPM are perceived benefits, perceived barriers, self-efficacy, activity-related affect, interpersonal influences, situational influences, commitment to a plan of action, and immediate competing demands. In Taiwan, 969 middle school students (55% males; 45% females) from Taipei provided data for the study. In the United States, the sample was collected from 286 late elementary and middle school students (48% males; 52% females). Results showed the gender differences in activity levels were apparent in the youths from both countries. In both Taiwanese and American youths, boys were more active than girls. Cross-cultural differences in the importance of barriers to physical activity emerged. Among Taiwanese adolescents, barriers did not emerge as a significant direct predictor of physical activity as they did among US adolescents. Perceived efficacy directly predicted physical activity among Taiwanese youths while it indirectly predicted physical activity and appeared to be mediated by beliefs regarding exercise benefits and barriers among American youths. In addition, the paths of effect for interpersonal influences were different when Taiwanese and US youths were compared. The findings from this paper have important and culturally relevant information that can inform future physical activity intervention studies with diverse adolescents. PMID- 11923902 TI - Correlations among serum calcium, vitamin D and parathyroid hormone levels in the elderly in southern Taiwan. AB - This study correlates serum vitamin D levels to related hormones and dietary intakes among 57 elderly Chinese above the age of 65 who were living in the same community in rural Southern Taiwan (Pingtung) and who had no conditions or drug intake known to interfere with the metabolism of vitamin D. Demographic characteristics, past medical history, medications, and dietary intake were collected via questionnaires. Venous blood samples were collected for analyses of serum 25-hydroxyvitamin D (25(OH)D), parathyroid hormone (PTH) and calcium levels. Our results showed subjects in this study to have normal mean values of serum 25(OH)D, PTH and calcium levels. The mean serum 25(OH)D level was 36.21 (+/ 6.37) ng/ml, the mean serum PTH level 29.24 (+/- 18.62) pg/ml and the mean serum calcium level 9.14 (+/- 0.52) mg/dl. While the mean serum 25(OH)D and calcium values were not found to be significantly different between men and women, the mean serum PTH level was significantly higher in women (33.42 +/- 20.00 pg/ml) than in men (23.07 +/- 14.66 pg/ml) (p <.05), and serum PTH levels were significantly negatively correlated to serum calcium (r = -.33, p <.05) but not 25(OH)D (r = -.21). A higher intake of calcium was significantly associated with higher serum calcium levels (r =.29, p <.05), but not with serum 25(OH)D levels. Results from this study suggested that the elderly people living in Pingtung, a particularly sunny region, had normal serum 25(OH)D levels. The fact that the elderly women studied had higher serum PTH levels and that these levels were negatively correlated to serum calcium levels suggests that a higher PTH level in the elderly women may be related to susceptibility for osteoporosis. In an effort to provide optimal nursing care for the elderly by minimizing hip fractures and related morbidity, further nursing studies are needed to study the effects of the environment, dietary intake and bone metabolism. PMID- 11923903 TI - The correlation between perceptions of control and hope status in home-based cancer patients. AB - The purpose of this study was to analyze correlation between perceptions of control and status of hope in home-based cancer patients. One hundred thirty seven home-based cancer patients from two hospital-based oncology centers in Taipei were interviewed using a structured questionnaire. The results showed that subjects' perceptions of control and status of hope were at a moderate level. Patient's religion, marital status and stage of cancer were significantly related to their hope status. The greater the age and the more serious the symptoms, the lower the status of hope was. The healthier the patients, the higher the status of hope was. Perceptions of control for patients was positively correlated with status of hope, when patients felt more sense of control in their ill situation, they had a higher status of hope. The findings of this study suggest that it is advantageous to establish support groups for home-based cancer patients and their families, so as to improve perceptions of control and facilitate their hope status. In addition, the model of case management can alleviate hopelessness due to physical function changes and distress of symptoms. In the future, more research on how to increase perception of control in patients, especially on internal control, is necessary in order to make appropriate and specific care plans for cancer patients. PMID- 11923910 TI - A genomewide linkage scan for quantitative-trait loci for obesity phenotypes. AB - Obesity is an increasingly serious health problem in the world. Body mass index (BMI), percentage fat mass, and body fat mass are important indices of obesity. For a sample of pedigrees that contains >10,000 relative pairs (including 1,249 sib pairs) that are useful for linkage analyses, we performed a whole-genome linkage scan, using 380 microsatellite markers to identify genomic regions that may contain quantitative-trait loci (QTLs) for obesity. Each pedigree was ascertained through a proband who has extremely low bone mass, which translates into a low BMI. A major QTL for BMI was identified on 2q14 near the marker D2S347 with a LOD score of 4.04 in two-point analysis and a maximum LOD score (MLS) of 4.44 in multipoint analysis. The genomic region near 2q14 also achieved an MLS >2.0 for percentage of fat mass and body fat mass. For the putative QTL on 2q14, as much as 28.2% of BMI variation (after adjustment for age and sex) may be attributable to this locus. In addition, several other genomic regions that may contain obesity-related QTLs are suggested. For example, 1p36 near the marker D1S468 may contain a QTL for BMI variation, with a LOD score of 2.75 in two-point analysis and an MLS of 2.09 in multipoint analysis. The genomic regions identified in this and earlier reports are compared for further exploration in extension studies that use larger samples and/or denser markers for confirmation and fine-mapping studies, to eventually identify major functional genes involved in obesity. PMID- 11923911 TI - A genomewide scan for loci involved in attention-deficit/hyperactivity disorder. AB - Attention deficit/hyperactivity disorder (ADHD) is a common heritable disorder with a childhood onset. Molecular genetic studies of ADHD have previously focused on examining the roles of specific candidate genes, primarily those involved in dopaminergic pathways. We have performed the first systematic genomewide linkage scan for loci influencing ADHD in 126 affected sib pairs, using a approximately 10-cM grid of microsatellite markers. Allele-sharing linkage methods enabled us to exclude any loci with a lambda(s) of > or =3 from 96% of the genome and those with a lambda(s) of > or =2.5 from 91%, indicating that there is unlikely to be a major gene involved in ADHD susceptibility in our sample. Under a strict diagnostic scheme we could exclude all screened regions of the X chromosome for a locus-specific lambda(s) of >/=2 in brother-brother pairs, demonstrating that the excess of affected males with ADHD is probably not attributable to a major X linked effect. Qualitative trait maximum LOD score analyses pointed to a number of chromosomal sites that may contain genetic risk factors of moderate effect. None exceeded genomewide significance thresholds, but LOD scores were >1.5 for regions on 5p12, 10q26, 12q23, and 16p13. Quantitative-trait analysis of ADHD symptom counts implicated a region on 12p13 (maximum LOD 2.6) that also yielded a LOD >1 when qualitative methods were used. A survey of regions containing 36 genes that have been proposed as candidates for ADHD indicated that 29 of these genes, including DRD4 and DAT1, could be excluded for a lambda(s) of 2. Only three of the candidates-DRD5, 5HTT, and CALCYON-coincided with sites of positive linkage identified by our screen. Two of the regions highlighted in the present study, 2q24 and 16p13, coincided with the top linkage peaks reported by a recent genome-scan study of autistic sib pairs. PMID- 11923912 TI - A combined analysis of genomewide linkage scans for body mass index from the National Heart, Lung, and Blood Institute Family Blood Pressure Program. AB - A combined analysis of genome scans for obesity was undertaken using the interim results from the National Heart, Lung, and Blood Institute Family Blood Pressure Program. In this research project, four multicenter networks of investigators conducted eight individual studies. Data were available on 6,849 individuals from four ethnic groups (white, black, Mexican American, and Asian). The sample represents the largest single collection of genomewide scan data that has been analyzed for obesity and provides a test of the reproducibility of linkage analysis for a complex phenotype. Body mass index (BMI) was used as the measure of adiposity. Genomewide linkage analyses were first performed separately in each of the eight ethnic groups in the four networks, through use of the variance component method. Only one region in the analyses of the individual studies showed significant linkage with BMI: 3q22.1 (LOD 3.45, for the GENOA network black sample). Six additional regions were found with an associated LOD >2, including 3p24.1, 7p15.2, 7q22.3, 14q24.3, 16q12.2, and 17p11.2. Among these findings, the linkage at 7p15.2, 7q22.3, and 17p11.2 has been reported elsewhere. A modified Fisher's omnibus procedure was then used to combine the P values from each of the eight genome scans. A complimentary approach to the meta-analysis was undertaken, combining the average allele-sharing identity by descent (pi) for whites, blacks, and Mexican Americans. Using this approach, we found strong linkage evidence for a quantitative-trait locus at 3q27 (marker D3S2427; LOD 3.40, P=.03). The same location has been shown to be linked with obesity-related traits and diabetes in at least two other studies. These results (1) confirm the previously reported obesity-susceptibility locus on chromosomes 3, 7, and 17 and (2) demonstrate that combining samples from different studies can increase the power to detect common genes with a small-to-moderate effect, so long as the same gene has an effect in all samples considered. PMID- 11923915 TI - [Masticatory muscles. Part II. Functional properties of the masticatory muscle fibers]. AB - The masticatory muscles in human beings and in other mammals show a number of specific adaptations. Their muscle fibres contain at least four different isoforms of myosin heavy chain (MHC) and many fibres express more than one kind simultaneously. This implicates a continuous range of fibre contraction speeds for these muscles. Most or all fibres have a high oxidative capacity and consequently are very resistant against fatigue. The fast muscle fibres of the masticatory muscles appear to have smaller calibers than the slow ones. This condition is particular and points to mild atrophy. The motor units of the jaw muscles are characterised by large fibre numbers per unit, but concentrated into small subvolumes of muscle. Because of the anatomical heterogeneity of the muscles, motor units are capable of generating quite a variety of force directions. The fibres of jaw muscle motor units often belong to different fibre types, as far as their MHC-composition is concerned. For this reason, the units cannot be subdivided into clear-cut types, but show a continuous range of contraction times. PMID- 11923914 TI - Generalized T2 test for genome association studies. AB - Recent progress in the development of single-nucleotide polymorphism (SNP) maps within genes and across the genome provides a valuable tool for fine-mapping and has led to the suggestion of genomewide association studies to search for susceptibility loci for complex traits. Test statistics for genome association studies that consider a single marker at a time, ignoring the linkage disequilibrium between markers, are inefficient. In this study, we present a generalized T2 statistic for association studies of complex traits, which can utilize multiple SNP markers simultaneously and considers the effects of multiple disease-susceptibility loci. This generalized T2 statistic is a corollary to that originally developed for multivariate analysis and has a close relationship to discriminant analysis and common measure of genetic distance. We evaluate the power of the generalized T2 statistic and show that power to be greater than or equal to those of the traditional chi2 test of association and a similar haplotype-test statistic. Finally, examples are given to evaluate the performance of the proposed T2 statistic for association studies using simulated and real data. PMID- 11923913 TI - Genetic dissection of the human leukocyte antigen region by use of haplotypes of Tasmanians with multiple sclerosis. AB - Association of multiple sclerosis (MS) with the human leukocyte antigen (HLA) class II haplotype DRB1*1501-DQB1*0602 is the most consistently replicated finding of genetic studies of the disease. However, the high level of linkage disequilibrium (LD) in the HLA region has hindered the identification of other loci that single-marker tests for association are unlikely to resolve. In order to address this issue, we generated haplotypes spanning 14.754 Mb (5 cM) across the entire HLA region. The haplotypes, which were inferred by genotyping relatives of 152 patients with MS and 105 unaffected control subjects of Tasmanian ancestry, define a genomic segment from D6S276 to D6S291, including 13 microsatellite markers integrated with allele-typing data for DRB1 and DQB1. Association to the DRB1*1501-DQB1*0602 haplotype was replicated. In addition, we found that the class I/extended class I region, defined by a genomic segment of approximately 400 kb between MOGCA and D6S265, harbors genes that independently increase risk of, or provide protection from, MS. Log-linear modeling analysis of constituent haplotypes that represent genomic regions containing class I (MOGCA D6S265), class III (TNFa-TNFd-D6S273), and class II (DRB1-DQB1) genes indicated that having class I and class II susceptibility variants on the same haplotype provides an additive effect on risk. Moreover, we found no evidence for a disease locus in the class III region defined by a 150-kb genomic segment containing the TNF locus and 14 other genes. A global overview of LD performed using GOLD identified two discrete blocks of LD in the HLA region that correspond well with previous findings. We propose that the analysis of haplotypes, by use of the types of approaches outlined in the present article, should make it possible to more accurately define the contribution of the HLA to MS. PMID- 11923916 TI - [Variations among dentists in the diagnosis of caries and assessment of dental restorations]. AB - It is well-known that dentists differ in judgement when confronted with the same clinical situation. To study this phenomenon, 2 groups of 5 general dental practitioners have assessed the material of 20 patients, each consisting of a set of 2 bite wing radiographs and 2 photographs of the (pre)molar region. The teeth were examined for caries and the quality of the restorations, supported by a protocol which was mailed in advance. The examiner variability was expressed as Cohen's kappa, calculated per pair of GDPs and subsequently averaged per group. The interexaminer agreement was fair for the diagnosis of caries and was highest for the assessment of approximal caries on the bite wing radiograph. The agreement with respect to the quality assessment of dental restorations proved to be poor on average. The intra-examiner agreement varied widely between examiners. No difference could be established between the groups. It can be concluded that, without training and calibration, a sufficient inter- and intra-examiner agreement among GDPs is difficult to reach. Consequently, a substantial variation is found in the individual GDP's restorative treatment planning. PMID- 11923917 TI - [Relation between mercury and Alzheimer's disease?]. AB - Mercury from dental silver amalgam has been suggested to be linked with Alzheimer's disease. The percentage in the population (formerly) possessing amalgam restorations exceeds that of Alzheimer patients. Therefore, low levels of mercury in the brain are unlikely to cause the disease. However, studies indicating increased levels of mercury in Alzheimer brains compared to controls have been criticised. In contrast, it has been proven that an inherited trait accounts for a minor proportion of Alzheimer's patients. Moreover, education--or related psychosocial factors--instead of amalgam has been found to be associated with the disease. Therefore, it does not seem warranted as yet to conclude that mercury is an etiological factor in Alzheimer's disease. PMID- 11923918 TI - [Left-handedness in dental undergraduates and orthodontic specialists]. AB - A questionnaire was devised involving a group of dental students (n = 70) and a group comprising all consultant orthodontists in the UK (n = 170) to investigate the prevalence and the role of handedness in dental specialisation. Subjects were classified as being pure left-, mixed- or pure right-handed according to responses to a hand preference questionnaire and the results were compared with a very similar previous study of the general population. The prevalence of sinistrality (classified by writing) was recorded as 8.6% among dental students and 17.2% among orthodontists; this compares with 7.4% among the general population. More mixed-handers presented in both the dental groups compared to the general population. This agreed with the right shift theory of laterality. No significant correlation was noted between handedness and any other variable between the two dental groups. PMID- 11923919 TI - [Herpetic gingivostomatitis on the basis of infection with HSV type 2]. PMID- 11923920 TI - [Globalization of health. II]. PMID- 11923921 TI - [Sjogren's syndrome. Consequences for oral health]. AB - Sjogren's syndrome is an auto-immune disease involving exocrine glands causing amongst others xerostomia and dry eyes. Many other tissues may be affected as well. Early recognition of this disease may help to alleviate signs and symptoms and is, thus, important. The aim of this paper is to provide the dentist with knowledge to enable him to timely diagnose the oral component of Sjogren's syndrome as well as to give guidelines for the treatment. PMID- 11923922 TI - [Masticatory muscles. Part VI. Masticatory muscles and movement of the lower jaw]. AB - The movement paths of the kinematic center of the temporomandibular joint were recorded by means of a jaw movement recording system (OKAS-3D) under 3 conditions: 1. free open and close movements; 2. free opening and loaded closing movements (subjects closed against a small, manually applied, downward directed force on the chin); and 3. during chewing of chewing gum. During free jaw movements, the opening path of the kinematic center lies above the closing path. During loaded closing movements, the opening and closing paths coincide. This indicates that during opening and loaded closing, the condyle-disc complex is slightly pressed against the articular eminence. However, during free closing, there is more space between the articulating surfaces. During gum chewing, the opening and closing paths of the condyle coincide on the balancing side, on the working side they don't. Thus, the joint on the balancing side is loaded and the joint on the working side is not. PMID- 11923923 TI - [Serious periodontal destruction in the elderly. Intensive treatment or an overdenture prosthesis?]. AB - A minority of dentate elderly people have advanced periodontal disease. Is both non-surgical and surgical periodontal treatment or overdenture treatment the therapy of choice? Research data comparing these treatments, from patients' or professionals' point of view, are as yet not available. Concerning efficacy and cost-effectiveness, overdenture treatment appears to be superior to periodontal treatment. PMID- 11923925 TI - [Over the limits... an analysis of dental care delivery]. PMID- 11923924 TI - [Occlusion repair or occlusion loss? Results of preliminary treatment as the starting point]. AB - Epidemiological data confirm that the number of elderly with an impaired dentition is increasing. The treatment of patients with an impaired dentition should be focused on maintaining oral function during life-time. However this goal is hardly achievable because of the complicating factors in an impaired dentition. This article describes the Nijmegen Dental School approach in the treatment of impaired dentition and stresses the importance of the preliminary treatment in order to answer the question whether restoring or extracting teeth is the most appropriate treatment modality. PMID- 11923926 TI - [Bite force and the force of movement in the masticatory system]. PMID- 11923927 TI - [Complaints caused by palladium-containing alloys]. PMID- 11923928 TI - [Dutch public health in the future I]. PMID- 11923929 TI - Transmissible spongiform encephalopathies: the viral concept and an application. PMID- 11923930 TI - Bovine spongiform encephalopathy and its relationship to the new variant form of Creutzfeldt-Jakob disease. An account of bovine spongiform encephalopathy, its cause, the clinical signs and epidemiology including the transmissibility of prion diseases with special reference to the relationship between bovine spongiform encephalopathy and the variant form of Creutzfeldt-Jakob disease. PMID- 11923931 TI - Animal transmissible spongiform encephalopathy: clinical and diagnostic aspects. PMID- 11923932 TI - The challenge for the public health system. PMID- 11923933 TI - [URE3] and [PSI]: prions of Saccharomyces cerevisiae. PMID- 11923934 TI - Structural biology of prions. PMID- 11923935 TI - Strain variations and species barriers. PMID- 11923936 TI - Resistance of transmissible spongiform encephalopathy agents to decontamination. PMID- 11923937 TI - Human prion diseases: cause, clinical and diagnostic aspects. PMID- 11923938 TI - The prion theory: background and basic information. PMID- 11923939 TI - Epidemiology and risk factors of transmissible spongiform encephalopathies in man. PMID- 11923940 TI - Patient-physician relationships in the information age. AB - The ready and free availability of information that characterizes the Information Age--with health information on the Internet and direct-to-consumer advertising of prescription drugs as two of these major information sources--is bringing about significant changes in patient-physician relationships. By developing a matrix of intended and unintended consequences and desirable and undesirable consequences, it's possible to have a better understanding of the impact of the free flow of information on this traditional relationship. Ultimately, any marketing approach to these newly empowered patients must take their level of health care knowledge into consideration. PMID- 11923941 TI - Assessing the marketing and development of health research organizations. PMID- 11923942 TI - Dimensions of the ideal HMO brand. PMID- 11923943 TI - Forecasting U.S. health care's future. PMID- 11923944 TI - Capturing and retaining more referral sources. PMID- 11923945 TI - [Selective modulators of estrogen receptors]. PMID- 11923946 TI - [Occupational medicine in the 19th century]. PMID- 11923947 TI - [Classification of systemic vasculitides]. AB - Several classifications of systemic vasculitides have been proposed: the Chapel Hill nomenclature, the classification criteria of the American College of Rheumatology and Lie's classification. All have their advantages and disadvantages. The role of ANCA (antineutrophil cytoplasmic antibodies) in classification of vasculitides is now accepted. The classification systems and the contribution of ANCA have led to better distinguishing vasculitis of medium calibre vessels from that involving small vessels, and in particular to distinguish between periarteritis nodosa and microscopic polyangiitis. Polyarteritis nodosa is characterised by involvement of medium calibre vessels (which can lead to vascular nephropathy or other ischemic organ involvement), whereas microscopic polyangiitis can be responsible, among other signs, for glomerulonephritis and pulmonary capillaritis. PMID- 11923949 TI - [Periarteritis nodosa]. AB - Periarteritis nodosa is a systemic vasculitis involving arteries with a diameter greater than 70-150 microns. Large vaccination against hepatitis B virus explains the decreased frequency of its association with hepatitis B virus observed in less than 10% of cases. Weight loss, fever, livedo, myalgias or arthralgias, mononeuropathy, gastrointestinal tract involvement, renal insufficiency, and cardiac failure are the most frequent clinical manifestations. Inflammatory syndrome and increased white blood cell count are common. Renal or digestive microaneurysms are present in more than 60% of cases. Prognosis is dependent of a five factors score corticosteroids with or without immunosuppressive drugs, result in 5-year survival rate over 60%. In hepatitis B virus-related periarteritis nodosa, corticosteroids are rapidly stopped; plasma exchanges in combination with antiviral agents (interferon alpha or lamivudine) give excellent results. PMID- 11923948 TI - [Causes and mechanisms of vasculitides]. AB - Vasculitic lesions are defined by necrosis of the vascular wall with perivascular inflammatory infiltrates. Secondary forms of vasculitis are observed in some systemic diseases, after infections, secondary to drugs or to malignancies. Usually, these secondary forms are characterised by the presence of immunoglobulin and complement deposits and thus probably caused by immune complex deposits. Conversely, necrotizing vasculitis occurring without known causal factor are named primary vasculitis, are not associated with immune deposits. Wegener's granulomatosis, microscopic polyangitis, Churg and Strauss syndrome and the isolated form of necrotizing and crescent glomerulonephritis are closely associated with the anti-neutrophil cytoplasm auto-antibodies (ANCA). The two main auto-antigen targets recognised by ANCA are lysosomal enzymes, proteinase 3 and myeloperoxidase, both contained in azurophilic granules of polymorphonuclear neutrophils and in primary lysosomes of monocytes. It has been demonstrated that ANCA antigens are expressed at the neutrophil (and monocyte) membrane after preactivation and are accessible to antibodies. Thus, ANCA may amplify neutrophils and monocyte activation. However, the causal or induced nature of anti-neutrophil auto-immunity in vasculitis remains unknown. PMID- 11923950 TI - [Microscopic polyangiitis]. AB - Microscopic polyangiitis is a systemic necrotizing vasculitis affecting small vessels, without granulomata. Patients present with rapidly progressive glomerulonephritis, often associated with cutaneous, musculoskeletal, neurological or gastrointestinal involvement. Antineutrophilcytoplasmic antibodies are frequently found, most often with a perinuclear pattern and a antimyeloperoxidase specificity. Diagnosis is based on the evidence of a necrotizing glomerulonephritis, or histological vasculitis, in the absence of extravascular granulomata or sign of macroscopic polyarteritis nodosa. Corticosteroids and immunosuppressive drugs have improved the prognosis, but relapses are frequent. PMID- 11923951 TI - [Wegener's granulomatosis]. AB - Wegener's granulomatosis is characterized by 3 major lesions: necrosis, granulomatosis, and vasculitis. Although all organs may be involved, chronic rhinosinusitis, pulmonary involvement (especially with the typical imaging pattern of cavitary nodules), and glomerulonephritis are the most common clinical manifestations. Antineutrophil cytoplasmic autoantibodies (characteristically cytoplasmic with anti-proteinase 3 specificity) are of considerable help for diagnosis. Remission of disease is obtained with corticosteroids and cyclophosphamide, but relapses are frequent, and treatment toxicities are a major concern. PMID- 11923952 TI - [Mixed cryoglobulinemia]. AB - Cryoglobulins are immunoglobulins that persist in serum, precipitate with cold temperature, and resolve when rewarmed. Cryoglobulinemia are part of systemic vasculitis, and they are in the subgroup of immune complex type vasculitis. Mixed cryoglobulinemia are associated with malignant B lymphoproliferative disorders, auto-immune and chronic infectious diseases, particularly hepatitis C virus infection. Mixed cryoglobulinaemia has a variable prognosis, depending mainly on renal involvement, systemic extension, and severity of hypertension. In pilot studies, alpha interferon seemed promising to treat symptomatic mixed cryoglobulinemia, because of its immunomodulating and antiviral properties. In severe or resistant forms of mixed cryoglobulinemia, plasmapheresis or immunosuppressive agents may be useful. PMID- 11923953 TI - [Rheumatoid purpura]. AB - Schonlein-Henoch purpura is a clinical syndrome characterized by the association of skin, joint and gastrointestinal symptoms consisting respectively in purpura, arthralgia and abdominal pain. The incidence of renal symptoms ranges from 33 to 50%. The long term prognosis is related to the renal disease which is characterized by a glomerular involvement, with hematuria and sometimes proteinuria. In more severe cases, a nephrotic syndrome is present with a decrease in renal function. A renal biopsy is performed when proteinuria is in excess of 1 g/day. It shows a mesangial proliferation and in more severe cases extracapillary proliferation while mesangial IgA deposits are seen by immunofluorescence. Severe forms require a treatment with corticosteroids, started with methylprednisolone pulses. Treatment should be started early in the course of the disease before glomerular crescents become fibrous. PMID- 11923955 TI - [Data, projections, and perspectives of doctor Cooper]. PMID- 11923954 TI - [Treatment of systemic vasculitides]. AB - Systemic vasculitis, which had been a consistently fatal disorder, became curable, with a sometimes chronic progression, when treatment with corticosteroids in association with immunosuppressive drugs was introduced. Treatment must be adapted according to causative factors, confirmed prognostic indicators, risk of recurrence and the need to reduce to a minimum iatrogenic side effects. Treatment of periarteritis nodosa, not linked to hepatitis B virus or to the Churg and Strauss syndrome, is based on high-dose corticosteroid treatment, associated with cyclophosphamide in intravenous bolus in severe forms. Microscopic polyangiitis and Wegener's granulomatosis usually require treatment with immunosuppressive drugs. These vasculitides have a high rate of recurrence and are responsible for high morbidity. Vasculitides associated to viral infections should receive specific treatment: plasma exchange and antiviral treatment (interferon, lamivudin) for periartiritis nodosa with hepatitis B, and interferon and ribavirin for cryoglobulinaemia associated with hepatitis C virus. PMID- 11923956 TI - [Lower limb edema. Diagnostic approach]. PMID- 11923957 TI - [Acute delirium status. Diagnostic approach and management in emergency situations]. PMID- 11923958 TI - [Epidemiology of transmissible diseases. Surveillance methods, notification, research on an outbreak in a community and preventive measures]. PMID- 11923959 TI - [Acute hypertensive crisis. Diagnosis and management in emergency situations]. PMID- 11923960 TI - [Purpura. Diagnostic approach]. PMID- 11923961 TI - [Angina. Etiology, diagnosis, course, treatment]. PMID- 11923962 TI - [Typhoid and paratyphoid fever. Epidemiology, etiology, physiopathology, diagnosis, course, treatment]. PMID- 11923963 TI - [Neurologic complications attributable to alcohol use. Diagnosis, treatment]. PMID- 11923964 TI - Revolutionary change may be next for a tired AMA. With dues declining, doc group could collect 'tithe' as umbrella organization for specialty, medical societies. PMID- 11923965 TI - Buying spree. New investor-owned start-ups adding hospitals. PMID- 11923966 TI - Deja vu all over again. N.J. malpractice insurer under fire after huge loss. PMID- 11923967 TI - One-stop shopping. Forget the typical gift shop. As retail options soar, patients might think they're at the mall, not the community hospital. PMID- 11923968 TI - Building the brand. Construction & design survey shows healthcare's focus on expanding specialized services. PMID- 11923969 TI - Ready, aim, litigate. AHA takes feds to court to stop HHS from eliminating Medicaid loophole. AB - The American Hospital Association and several other groups are doing their best to stop HHS from eliminating the upper payment limit loophole, and the AHA is even suing HHS, but time is running out for the payment plan worth $27 billion over the next 10 years to State Medicaid programs. This is only the fourth time in the past decade that the AHA has sued the federal government. PMID- 11923970 TI - A new chapter in my life. PMID- 11923971 TI - WORLD welcomes new executive director. PMID- 11923972 TI - HIV and pregnancy: treatment guidelines updated. PMID- 11923973 TI - Reducing the risks of conception. PMID- 11923974 TI - Our first line of defense against bioterrorism. Part 1. AB - Health care's response to the anthrax outbreak highlights our critical need to enlist sophisticated information technology (IT) tools in the defense against bioterrorism. Part 1 of a two-part series explains what went wrong in the most recent attack and suggests how a national IT infrastructure might help in the future. PMID- 11923975 TI - Honey, it's about attitude. PMID- 11923976 TI - Unravel pain standards for higher compliance. AB - Since implementation in 2000, the Joint Commission has made minor--yet intricate- changes to its new pain standards to gain higher compliance. PMID- 11923977 TI - Expert witnessing brings nursing expertise into the legal arena. AB - Get the basics regarding nurses and expert witnessing. PMID- 11923978 TI - Past victim, future victim? AB - To effectively reduce and prevent workplace violence, nurse managers can offer training sessions that include assessing risk for past and current victimization. PMID- 11923979 TI - Survival. AB - A 500-bed hospital enlists more than one-third of its staff to prepare for a Joint Commission survey. PMID- 11923980 TI - Let's bring balance to health care. AB - Consider implementing the balanced scorecard--a performance management method that allows you to view your work from four perspectives: customer, internal business, learning and growth, and financial. PMID- 11923981 TI - Rework the workload. AB - Kindred Healthcare, Inc., the nation's largest full-service network of long-term acute care hospitals, initiated a 3-year strategic plan to re-evaluate its workload management system. Here, follow the project's most important and difficult phase--designing and implementing the patient classification system. PMID- 11923982 TI - One heart, countless paths. AB - One of the many things I love about the nursing profession is the immeasurable number of roles it offers us. Clinical options exist in acute care, long-term care, ambulatory care, school nursing, home care, and hospice care, to name a few. If management and research "call" you, you can pursue your passion in health care delivery systems, clinical research institutes, the pharmaceutical industry, or in any number of medical device and technology companies. PMID- 11923983 TI - Beyond the psychomotor realm. AB - Enhance the accuracy and continuity of your competence assessment by defining nursing skills with the research-based Nursing Interventions Classification. PMID- 11923984 TI - Data sparks new care model. AB - Nurses at Hennepin County Medical Center (HCMC), a public teaching hospital in Minneapolis, Minn., combined clinical data and critical thinking savvy to create positive change. PMID- 11923985 TI - Monitoring vital signs in vital times. AB - Vital signs monitoring advances make it possible for caregivers to assess patients with the same equipment, from admission until discharge. Before purchasing a monitoring system, be sure to clearly understand your patients' short- and long-term monitoring needs throughout the hospital. PMID- 11923986 TI - A new radiology resource. PMID- 11923987 TI - Saving Frank. PMID- 11923988 TI - Magnet hopitals: A key to reducing the nursing shortage. PMID- 11923989 TI - A chilling reality. PMID- 11923990 TI - Survey of instruments. Glucose analyzers 2002--sweeter than ever. PMID- 11923991 TI - Analysis of quality of life data in controlled clinical trials. PMID- 11923992 TI - Simultaneous analysis of quality of life and survival data. AB - In many phase III clinical trials, particularly in the field of cancer, the comparison of treatments is based on both length of survival and quality of life. Subjects are followed over time until death and during this period, quality of life is assessed on a number of occasions. Simultaneous analysis of these two outcomes supplements the comparison of treatments in terms of each outcome independently with an assessment of the net effect. In addition, it provides a means of accounting for the informative dropout due to death of patients within the time frame of the quality of life study. The methods also have the potential to be extended to allow for informative dropout from the quality of life study prior to death. There are a number of broad approaches for the simultaneous analysis of quality of life and survival data. The most widely used approach in clinical research is quality-adjusted survival analysis, where treatments are compared in terms of a composite measure of quality and quantity of life. The paper reviews the different techniques for quality-adjusted survival analysis, illustrating the methodology by application to data from a phase III clinical trial in pancreatic cancer. In addition, alternative approaches using multistate survival analysis and joint modelling methods are also discussed. PMID- 11923993 TI - A review of ordinal regression models applied on health-related quality of life assessments. AB - There has been increasing emphasis in medical research on the design and analysis of quality of life scales. Many quality of life scales are ordinal and statistical methods such as ordinal regression models have been reviewed on a number of occasions. However, when such models are applied, the way the data have been generated is often overlooked. In this paper we illustrate the use of ordinal regression models, in particular the proportional odds model, the partial proportional odds model and the stereotype model in the MRC Cognitive Function and Ageing Study (MRC CFAS). The partial proportional odds and the stereotype models are often under-utilized, perhaps due to the lack of available software. However, in this paper, analysis based on these models has been carried out using the popular statistical software package SAS and macros devised in SAS. Furthermore, bootstrapping techniques have been applied to obtain valid estimates of the standard errors of the parameters in the stereotype model. Strikingly different results were obtained using the different ordinal regression models. We conclude that the way the data have been generated is particularly important for the analysis of quality of life assessments. Different methods of generating scores yield data with different properties. It is now possible to fit a variety of ordinal regression models and so select the appropriate one that correctly models the data. PMID- 11923994 TI - Analysing longitudinal continuous quality of life data with dropout. AB - Quality of Life (QL) is becoming an increasingly popular endpoint in phase III cancer clinical trials. However, there is still no agreement as to what is the optimal approach to analysis. In this paper we review some concepts which should be considered during a QL analysis. We present two modelling approaches that have been substantively developed in other research fields: selection models and pattern-mixture models. These models are compared using data from an EORTC clinical trial in poor-prognosis prostate cancer patients. It is illustrated that, although selection models and pattern mixture are probabilistically equivalent, they may shed completely different light on data from a modeller's point of view. PMID- 11923995 TI - The analysis of repeated multivariate binary quality of life data: a hierarchical model approach. AB - Many quality of life measuring instruments consist of a number of questions that are answered on ordinal scales. Often these responses are then totalled to give a summary score for each quality of life domain within the instrument. This, however, may lose valuable information about individual aspects of patient quality of life and also can have little intuitive meaning. Here we present an alternative analysis, in which dichotomized individual items of the questionnaire are analyzed. We first show how a hierarchical logistic regression model for repeated binary data can be extended to the multivariate case. We then use such a model for analyzing the prevalence of six symptoms in a palliative treatment trial in non-small-cell lung cancer. The analysis provides information about the correlations between symptoms, both between and within person. If appropriate, it also permits the estimation of a treatment effect common to all symptoms. Methods for model checking are discussed. We conclude that this methodology can provide a more intuitive and informative analysis of quality of life data than that obtained by considering summary scores. PMID- 11923996 TI - Infant holding policies and practices in neonatal units. AB - PURPOSE: To describe neonatal unit policies and practices regarding the holding of infants by parents. DESIGN: U.S. national cross-sectional descriptive survey. SAMPLE: Nurses representing 215 neonatal units providing regional-level (22 percent), community-level (45 percent), and intermediate-level (33 percent) care. MAIN OUTCOME VARIABLE: Proportion of respondents who reported neonatal intensive care unit criteria permitting parents and family members to hold their infant by conventional and skin-to-skin methods. RESULTS: All but one of the neonatal units offered parents the chance to hold their extubated infants conventionally; 73 percent offered parents to hold their extubated infants skin-to-skin. Sixty-four percent of neonatal units offered parents the option to hold their intubated infants conventionally. In contrast, only 45 percent of the units offered parents to hold their intubated infants skin-to-skin. Factors that influenced parental holding practices included potential benefits to infants and parents, concerns about infant safety, and level of support from neonatal staff members. PMID- 11923997 TI - Transition to the neonatal nurse practitioner role: making the change from the side to the head of the bed. AB - Role transition is never easy, but is complicated by the experienced neonatal nurse's frustration with reverting to a student role and becoming a novice practitioner, sometimes after years of developing a reputation as an expert nurse. This article discusses this transition, focusing on the skills needed to move successfully from nurse to nurse practitioner. Common to all advanced practice transitions are stages similar to those Benner identifies in her novice to-expert theory of nursing practice. Feelings of frustration and inadequacy are common during the first year as an NNP. Studies focusing on role transition and role development suggest that a strong nursing identity is important for success in the NNP practice environment. Strategies to enhance the transition are discussed. PMID- 11923998 TI - Infant crying: nature, physiologic consequences, and select interventions. AB - This article describes the nature of infant crying, the physiologic events and changes associated with it, and appropriate nursing interventions for infant crying. A cry is a series of four movements that basically resembles a Valsalva maneuver. Documented immediate and long-term sequelae of crying include increased heart rate and blood pressure, reduced oxygen level, elevated cerebral blood pressure, initiation of the stress response, depleted energy reserves and oxygen, interrupted mother-infant interaction, brain injury, and cardiac dysfunction. Caregivers are encouraged to answer infant cries swiftly, consistently, and comprehensively. Kangaroo care is an efficient method for preventing, minimizing, and halting crying. Other interventions for crying include swaddled holding, a pacifier, sugar water, a sweet-tasting nonsucrose solution, heartbeat sounds, distraction by lullabies or mother's voice, rhythmic movement, and reduction of external stimuli. PMID- 11923999 TI - Research methodologies in neonatal nursing. PMID- 11924000 TI - The neonatal liver, Part 1: embryology, anatomy, and physiology. AB - The liver is the largest organ in the body and is critical to a number of metabolic, regulatory, and detoxification processes. These include the production of bile, metabolic processing of nutrients, synthesis and regulation of plasma proteins and glucose, and biotransformation of drugs and toxins. PMID- 11924002 TI - Neural cowpaths in the NICU. PMID- 11924001 TI - Dysphagia and oral feeding problems in the premature infant. PMID- 11924003 TI - [Antitumor immune response: what are the roles for gliomas?]. AB - Major advances achieved in immunology along the last decade have considerably improved our appraisal of the interactions between tumor cells and cells of the immune system (including lymphocytes), leading to the current development of immunotherapies as a new treatment strategy against cancer. Due to their localization in the brain, glioma are usually considered to arise in an immunoprivileged site, precluding immune intervention. Recent data challenge such a nihilism. Indeed, glioma infiltrating lymphocytes include T cell clonal expansions, suggesting a response against glioma antigens not yet identified. Furthermore, there is now cumulative evidence that activated T cells can traffic through the central nervous system and that T cells primed by astrocytoma cells in the periphery can recirculate and reach the brain to mediate their anti-tumor effects. Finally, multiple mechanisms by which glioma evade the immune response have been elucidated. However, there are still important enigmas, such as the nature of glioma antigens and the steps of antigen presentation in the brain. Responding to these crucial questions should facilitate the development of immunotherapies against brain tumors, avoiding deleterious auto-immune reactions. PMID- 11924004 TI - [Amyotrophic lateral sclerosis and animal models]. AB - Amyotrophic lateral sclerosis (ALS) is a progressive neurodegenerative disease which affects cortical, bulbar and spinal motoneurones. The cause of the disease, probably due to several factors, is still unknown and the survival delay of patients with ALS generally does not exceed 3-5 years. Animals models provide a unique opportunity to study pathological features and to evaluate potential therapeutic effects of news treatments. Natural disease models, neurotoxins or viral-induced models and more recently transgenic models with genetic anomalies mimicking those found in ALS patients have been extensively studied. This review summarizes the most relevant clinical and pathological advances issuing from these animal studies. PMID- 11924005 TI - [Long-term treatment of Alzheimer's disease: followup of a cohort of 255 patients treated with lacrine for four years]. AB - We describe the follow-up of a cohort of 255 Alzheimer's disease (AD) patients (81 males, 174 females) treated by tacrine during 4 years. We performed the survey of hepatic, cholinergic and general tolerance. Drug efficacy was measured by MMS examination on weeks 0, 18, 30, 52, 104, 156 and 208. A total of 190 patients (74.5 percent) were dropped out of this study, 75 (29 percent) for adverse events. We found 85 hepatic (33 percent), 79 cholinergic (31 percent), 31 (12 percent) neuropsychiatric and 72 general (28 percent) side effects. In term of drug efficacy we observed a global decline of 2.5 MMS points during the first year and 2 MMS points between W52 and W156. Tacrine's symptomatic efficacy, defined as the number of patients improved or stabilized at W30, was present in 50 patients (46 percent) among the 109 patients reaching W30. The intensity of symptomatic efficacy was expressed by a 2.7 MMS points increase in 37 patients improved on W30. The long term effects of Tacrine, measured by the MMS score at one year, showed a positive impact as the MMS was 2.5 points above the expected score in non treated AD patients. This study raises the practical problem of optimal cholinesterase inhibitors use in AD and the theoretical question of long term action of cholinesterase inhibitors on cerebral lesions of AD. PMID- 11924006 TI - [IContribution of lexical recall in the Set Test in Alzheimer disease screening]. AB - We administered the Set Test to 128 control subjects and 82 patients with Alzheimer's disease. The subjects were to produce, within 15 seconds, items within specific categories: colors, animals, fruits and cities. Control subjects obtained a mean score of 40.3 +/- 3.6 and Alzheimer's disease patients a mean score of 2.7 +/- 4. We found a significant effect of item category, age and cultural level only in control subjects. There was an interaction between sex and item category in both groups. We observed a significant effect of the level of dementia as measured with the MMS only in the Set Test. The Set Test appears to require more attentional resources than fluency within two minutes. PMID- 11924007 TI - [Presenting a battery for assessing spatial neglect. Norms and effects of age, educational level, sex, hand and laterality]. AB - The aim of this study was to build up a battery for assessing spatial neglect, then to analyse the norms and potential effects of age, education level, sex, hand used, and laterality. It was also to contribute evaluating the pseudoneglect phenomenon described by Heilman, which consists in a tendency of normal subjects to neglect the right peripersonal space. Tasks selected were presented to important groups of normal subjects, most often larger than 450. The battery comprised of a bell cancellation test, scene copy, clock drawing, two line bisection tasks, identification of overlapping figures, text reading, writing task, and the representational task of the France map. For each of them, different variables were selected, especially investigating the difference between performance in the right and the left hemispaces. This study allowed defining the threshold values (percentiles 5 and 95) for deciding of the pathological character of a patient performance. It also showed that the pseudoneglect phenomenon is more obvious in some tasks such as line bisection, and probably also in the representational task of the France map and writing. His importance and at times his side were influenced by the factors we studied, with between tasks differences, but also by the nature of the task to be performed, and especially his verbal component. PMID- 11924009 TI - Associations between a cerebellar motor dysfunction scale and movement initiation and completion in olivopontocerebellar atrophy. AB - Correlations were drawn between subscores of a cerebellar motor dysfunction (CMD) scale and simple reaction time (RT) and movement time (MT) in patients diagnosed with olivopontocerebellar atrophy (OPCA). Poor performance on CMD subscores was correlated with long disease duration and lengthening of visual and auditory RT and MT, with the motor speed subscore showing the highest correlations. OPCA patients with severe cerebellar HMPAO uptake deficiency on single photon emission computed tomography scans had poorer performances on three of the four subscores (static equilibrium, dynamic equilibrium and nystagmus, and motor speed) than patients with mild deficiency. These results are discussed in terms of the role of cerebellar afferent/efferent pathways on different types of motor dysfunction in patients with cerebellar atrophy. PMID- 11924008 TI - [Hearing loss in multiple sclerosis: clinical, electrophysiologic and radiological study]. AB - Clinical presentation of multiple sclerosis (MS) is highly heterogeneous. Deafness is considered to be rare. The aim of this study was to determine the frequency of acute deafness in MS and to evaluate prognosis. The occurrence of acute deafness was systematically noted in a cohort of 400 consecutive clinically definite MS patients. Acute deafness was observed in 14 patients (3.5 p. cent) with a mean age of 28 years (15 to 48) and after a mean of 2.3 years (0 to 10) of disease duration. In all but two patients, deafness was associated with disease exacerbations, highly suggestive of infratentorial lesions. MRI showed hyperintensities in the pons or the midbrain in 11 cases. Deafness disappeared in all cases excepted for one patient. After a mean follow-up of 7.2 years (1 to 17), all the patients remain in the relapsing-remitting form of the disease with a mean EDSS of 2.4 (0 to 4.5). Acute deafness is observed in 3.5 p. cent of MS cases and occurs more frequently during the first years of the disease. Although follow-up of a larger cohort is necessary, deafness could be of good prognostic factor in MS. PMID- 11924010 TI - [Postural and locomotor evaluation of normal pressure hydrocephalus: a case report]. AB - Generally, the diagnosis of idiopathic normal pressure hydrocephalus (NPH) is raised in elderly patients with an association of gait disturbances, cognitive impairment, urinary incontinence and widening of the ventricles. Gait disturbances appear as the predominant clinical symptom of NPH, and may precede other symptoms. The main objectives of this study were to determine the effect of ventriculoperitoneal shunt (VPS) on gait and to compare the results obtained in HPN with those obtained in Parkinson's disease (PD). The performance of a patient with NPH on specific postural and locomotion tasks was analyzed during three successive experimental sessions: the first one before VPS, the second one week after VPS and the third three months after VPS. The patient was instructed to walk at his natural speed and also to maintain postural control for one minute. Four gait parameters (mean velocity, stride length, cadency and step width) were calculated for each session to assess the subject's performance. In this NPH patient there was a significant improvement in gait performance before and after VPS. This improvement concerned mean velocity, stride length and step cadency but did not involve the width component which remained unchanged. Before VPS, the NPH patient walked slower and with a shorter stride length than a group of patients with PD. After VPS his performance was better than the PD group except for cadency. Step width remained longer for the HPH patient than in PD patients for all three sessions. From this study, it appears that gait in idiopathic NPH exhibits a specific temporospatial pattern distinct from that in PD or normal age matched controls. Changes in gait can be evaluated shortly after treatment of NPH. Gait disorders, excepting dynamic balance, may normalize as soon as one week after surgery. PMID- 11924011 TI - [Hemichorea, hemiballism disclosing non-ketotic hyperglycemia]. AB - Non ketotic hyperglycemia is a metabolic disorder which can induce various hyperkinetic disorders. We report the case of a 74-year old woman admitted to hospital because of choreo-ballic movements of her left hemibody. Laboratory tests revealed non-ketotic hyperglycemia. CT scan showed an increased density in the right putamen and head of caudate. Magnetic resonance imaging disclosed a hyperintense signal involving the right putamen on T1-weighted images. On T2 weighted images, there was a slight hyperintensity in the right putamen and head of caudate. Correction of the underlying hyperglycemia led to complete resolution of abnormal movements. Pathophysiology of this clinicoradiologic syndrome is briefly discussed. PMID- 11924012 TI - [Rapidly evolving diabetic mononeuritis multiplex. Favorable outcome after immunosuppressive treatment]. AB - A 61 year-old man with type 2 diabetes mellitus presented with an extremely rapid and aggressive mononeuritis multiplex. Four months after onset, he had severe postural hypotension and at least 6 cranial nerves and 4 somatic nerves were involved. Extensive work-up failed to discover any etiology for the neuropathy apart from diabetes. Treatment with corticosteroids, i.v. immunoglobulins and cyclosporin was followed by progressive but sustained improvement. This case and few other published ones suggest that some particularly aggressive forms of diabetic neuropathy have an immune mechanism and may be treated favorably with immunosuppressor drugs. PMID- 11924014 TI - [Hyperproteinemia at 65g/l in a probable case of meningo-radicular tuberculosis]. PMID- 11924013 TI - [Radicular pain revealing Hodgkin's disease]. AB - We report on a case of Hodgkin's disease, revealed in a 52 year-old woman by isolated neurological signs. Sciatica is an uncommon pattern of discovery that is particularly misleading. It may lead to a delayed diagnosis all the more prejudicial since it discloses an advanced stage of the disease. The neurological involvement is always associated with a bone lesion. This latter can be discovered at an early stage through magnetic resonance imaging. PMID- 11924015 TI - [Central pontine and extra-pontine myelinolysis]. PMID- 11924016 TI - [Evaluation and treatment of non-traumatic intracerebral hemorrhages]. PMID- 11924018 TI - [Physiopathology of idiopathic hypersomnia. Current studies and new orientations]. AB - In 1976 Bedrich Roth coined the term "idiopathic hypersomnia" and described two forms of the disease, one monosymptomatic, manifested only by excessive daytime sleepiness, and one polysymptomatic, characterized by excessive daytime sleepiness, nocturnal sleep of abnormally long duration and signs of "sleep drunkenness" on awakening. In comparison with that of narcolepsy, the pathophysiology of idiopathic hypersomnia remains poorly known. There are two main reasons for that: the absence of clinical and polysomnographic criteria pathognomonic or at least characteristic of the condition, as the cataplexies and the sleep onset REM periods of narcolepsy, and also the absence of a natural animal model comparable with the canine model of narcolepsy. The first investigations have stressed the frequent familial pattern of idiopathic hypersomnia. Later on biochemical assays have been performed in the CSF with results in favour of a dysfunction of noradrenergic systems. In the light of the two process model of sleep regulation in which sleep propensity is determined by a homeostatic process S and a circadian process C and of the later three-process model of regulation in which sleepiness/alertness are simulated by the combined action of a homeostatic process, a circadian process and sleep inertia, we suggest that idiopathic hypersomnia is not a pathological entity in itself, but rather the consequence of chronic sleep deprivation in very long sleepers. PMID- 11924017 TI - [Recommendations for the creation of neuro-vascular units]. PMID- 11924019 TI - [Disorders of arousal]. AB - There are three slow wave sleep (SWS) parasomnias: confusional arousal, sleepwalking and night terrors grouped together in arousal disorders because impaired arousal from sleep has been postulated as a cause of these disorders. The onset of these disorders at the beginning of the night in slow wave sleep (SWS) is a typical feature. Night sleep is characterised by a great sleep fragmentation specially in SWS, and by a very strong SWS intensity reflected by both an increase of slow wave activity (SWA) before the parasomnia and a slower decay of wave activity (SWA) during the night. An abnormal deep sleep associated with a high SWS fragmentation is responsible for the occurrence of arousal disorders parasomnias. PMID- 11924020 TI - [Sleep talking]. AB - Sleep talking is very common in the general population. Its prevalence remains stable from childhood through adulthood. Sleep talking is often associated with other parasomnias: sleep walking, sleep terrors or REM sleep behavior disorders. It may arise from either REM or non REM sleep, when associated with REM sleep it is more comprehensible and often associated with clear sentences and recall of sleep mentation. Sleep talking is a benign entity and does not require any treatment; however an exceptional organic cause or psychopathology should be suspected if the onset is late (after 25 years); if the mental content is too violent or too emotional. PMID- 11924021 TI - [Treatment of parasomnias]. AB - In this article the most frequent and interesting treatments for parasomnias are reviewed. Arousal disorders, REM sleep behavior disorders and nightmares, bruxism (which can occur in any sleep state) are considered. The pathophysiological mechanisms of parasomnias are unknown, explaining why there is rarely a specific treatment precipitating. Management depends on the patient's age, the frequency and intensity of the episodes, the familial structure, the presence of precipitating and risk factors, psychological disorders and the parasomnia itself. Different therapeutic approaches are described including (1), explanation and reassurance, (2) counselling to avoid precipitating factors such as irregular sleep patterns, alcohol or drug intake (3) strategies to avoid injury (4) psychological approaches such as behavioral therapies in some parasomnias such as nightmares or arousal disorders. However, very few controlled studies have been done making it difficult to provide recommendations based on controlled clinical trials. Pharmacologic treatments are rarely necessary except in REM sleep behavior disorders; they are most often needed for short term. PMID- 11924022 TI - The somatotropic axis and sleep. AB - We review the evidence suggesting that hypothalamic growth hormone (GH)-releasing hormone (GHRH) stimulates sleep and growth hormone secretion simultaneously. GHRH injected into the cerebral ventricles, systemic circulation or the preoptic region enhances non-REM sleep (NREMS) in rats, rabbits and mice, and GHRH administered systemically promotes NREMS in humans. GHRH may also stimulate REMS but this effect is indirect and requires the presence of GH. Inhibition of endogenous GHRH (antibodies, antagonist, somatostatin, high doses of GH or IGF-1) suppresses both NREMS and GH secretion. Mutant rats and mice with deficiencies of GHRH signaling, and transgenic mice with decreased GHRH production sleep less than normal animals. Hypothalamic GHRH mRNA and GHRH content display diurnal variations and change in response to sleep deprivation. The NREMS-promoting activity of GHRH is independent of GH and is mediated by the preoptic region. It is suggested that promotion of NREMS and stimulation of GH are parallel outputs of hypothalamic GHRH through which anabolic activities in the body are are synchronized to periods of sleep. PMID- 11924023 TI - [Melatonin and circadian rhythm]. AB - Melatonin is an indole hormone that is produced by the pineal gland, mainly at night, with a peak around 3.00 a.m. under normal environmental conditions. This endogenic secretion cycle is generated by the suprachiasmatic nuclei in response to the day/night alternation. Light either suppresses or entrains melatonin production according to the time of light exposure. Melatonin can be viewed as the "hand" of the internal clock and is regulated via the central nervous and sympathetic systems. Melatonin synchronizes biological cycles, particularly the temperature and sleep/wake cycles. Exogenous melatonin can influence the endogenous secretion of melatonin according to a phase response curve, an effect that provides a rationale for the use of melatonin to treat disorders of biological rhythms (rapid time-zone change syndrome, delayed sleep phase syndrome, desynchronization in blind subjects or shift workers, insomnia in the elderly). PMID- 11924024 TI - [Delayed wakefulness-sleep rhythm syndrome and melatonin. Synthesis of existing studies]. AB - Delayed sleep phase syndrome involves undesirable late bed times and arising times with extreme difficulties in falling asleep and in awakening at a desired clock time. These patients present a delayed circadian system. Chronotherapy and phototherapy are designed to have a training effect on the circadian system. Response to these treatments varies widely and depends on the patient's motivation and associated psychological disorders. Other treatments have been proposed with less evident results. The few studies testing the effect of melatonin in delayed sleep phase syndrome concern a small number of patients and present methodological drawbacks. It can be concluded from these studies however that exogenous melatonin influences endogenous secretion more than other secretion rhythms. The effect on sleep time is significant but clinically moderate. More studies are needed to examine the effect of exogenous melatonin as a treatment strategy in delayed sleep phase syndrome. PMID- 11924025 TI - [Advanced sleep phase syndrome]. AB - The Advanced Sleep Phase Syndrome (ASPS) is a sleep disorder characterized by an early sleep onset and early awakening without any disturbance of the sleep structure. The management of this disease requires clinical and laboratory investigations in an attempt to confirm the phase advance of body core temperature and melatonin rhythm. The use of light therapy, possibly associated with chronotherapy or melatonin intake has been proposed. The evolution is variable. Seven subjects, aged 15 to 72 were diagnosed in our sleep disorders unit by mean of sleep log, actigraphy, sleep and temperature recording. The sleep onset and sleep offset times were approximately the same according to sleep log, actigraphy and night polysomnography. The nadir of body core temperature was at 01:38 +/- 01:03. Two familial cases were identified of which 1 was investigated in constant routine condition with hourly blood sampling. An advanced phase of melatonin and cortisol was evidenced. The disease temporarily improved in 3 cases with light therapy and in one case with the association of light therapy and chronotherapy. These data show the difficulties of the management and the treatment of this rarely diagnosed disease. PMID- 11924026 TI - [Disorders of wakefulness and sleep in blind patients]. AB - The goal of this paper was to summarize three studies focused on sleep/wake disorders in blind subjects. The first study was an epidemiology survey performed in 1073 blind subjects in comparison with non-blind controls. The blind had more episodes of insomnia and free running rhythms. They also took more sleeping pills and complained of more daytime somnolence. The seriousness of the sleep disorders was related to the seriousness of the blindness. In the second study, 78 blind children were compared with seeing children. They had more insomnia and more parasomnias but there was not any more free running. Finally, polysomnography was performed in 26 free running blind subjects in comparison with 26 controls. Total sleep time and sleep efficiency were lower in the blind. Sleep latency was increased and REM sleep was disturbed (longer latency and percentage decreases). There was no difference concerning slow wave sleep. Factorial analysis showed that factors such as being born blind, having ocular prosthesis, being single or having children had no influence on sleep. Working did however have an influence. PMID- 11924027 TI - [Photoperiod phototherapy and wakefulness-sleep rhythm disorders]. AB - Bright light therapy is a recent physical treatment in chronodisabled situations. The most recognized indication is the seasonal affective disorder. However, any disease or dysfunction where a misalignment of sleep-wake and circadian rhythms may be suspected is a potential tool for this treatment. Analyses of the literature throughout the interpretation methods of the evidence based medicine indicate that bright light therapy, if not a standard, could be recommended in a number of circadian rhythm sleep disorders, mainly the delayed and advanced sleep phase syndromes. Time aspects are essential for the success of phototherapy. From this point of view, easy and practical technological means or methods, allowing to shape a Phase Response Curve in each individual to be treated, should be clear progress. A future extension of indications will also depend on the checking of essential hypotheses linking circadian and sleep-wake rhythms in diseases such as psychophysiological insomnia, multiple sclerosis, brain dysgeneses or dementias. At last, a non negligible advantage of bright light therapy appears to be its relative safety. PMID- 11924028 TI - [Respiratory disorders during sleep and myasthenia]. AB - A sleep apnea syndrome is described by several studies in patients with myasthenia gravis. Apnea and hypopnea are mainly not obstructive and occur predominantly in rapid eye movement (REM) sleep. They are associated with oxygen desaturation. Although the sleep apnea index is not correlated with myasthenia gravis severity, it becomes less important as myasthenia gravis improves. Risk factors for the development of sleep apnea in myasthenia gravis patients are age, restrictive pulmonary syndrome, diaphragmatic weakness and daytime alveolar hypoventilation. Sleep apnea are not related to a central cholinergic effect in myasthenia gravis caused either by anticholinesterase used to treat myasthenia gravis or by antibodies to muscle acetylcholine receptors. Indeed acetylcholine receptors in brain are antigenically distinct from acetylcholine receptors in skeletal muscle. Sleep apneas are more likely caused by peripheral mechanisms. Correlation between sleep apnea and total lung capacity as well as the importance of diaphragmatic weakness in myasthenic patients may explain their predominance in REM sleep and their reduction with clinical improvement in the myasthenia gravis. PMID- 11924029 TI - [Respiratory disorders during sleep in degenerative diseases of the brain stem]. AB - Sleep-disordered breathing may be present in patients with degenerative diseases affecting the brainstem. Indeed, this last structure contains the executive system of rapid eye movement (REM) sleep (tegmentum of the pons), of respiratory drive (medulla oblongata and pons) and motor neurons of upper airways dilators (fifth, seventh, ninth, tenth and twelfth cranial roots). Patients with Parkinson's disease suffer frequently from insomnia, partly caused by nocturnal motor disability, and from REM sleep behavior disorder. In 20 percent of the patients, excessive daytime sleepiness is caused by a sleep apnea syndrome, with a partly levodopa-dependent upper airway dysfunction. In 40 percent of the patients, sleepiness mimics a secondary narcolepsy and may be associated with hypnagogic hallucinations. During supranuclear palsy, REM sleep is progressively curtailed with rare sleep-disordered breathing. Patients with multiple systemic atrophy may present a nocturnal stridor caused by laryngeal palsy and benefit from tracheotomy or continuous nasal positive airway pressure. Seldom sleep and respiratory studies in genetic ataxic diseases suggest a normal respiratory drive, occasional diaphragmatic dysfunction and night hypopneas. During amyotrophic lateral sclerosis, the progressive loss of phrenic nerve leads to a diaphragmatic dysfunction, dyspnea and a lesser survival. Adequate ventilation is jeopardized during REM sleep with a consequent loss of this state. PMID- 11924030 TI - [Respiratory disorders during sleep in amyotrophic lateral sclerosis]. AB - Amyotropic lateral sclerosis is a severe degenerative disease of the nervous system. Progressive death of central and peripheral motor neurons leads to rapidly extensive paralysis of the four limbs, the bulbar region and the respiratory system. Sleep respiratory disorders is a useful sign to follow in these patients. Apnea is generally central rather than obstructive. Hypoventilation, particularly during REM sleep has been reported and causes nocturnal desaturation. These patients also appear to have a disrupted sleep pattern with a longer phase I and a shorter total duration of sleep. These nocturnal respiratory disorders observed in amyotropic lateral sclerosis are similar to those observed in muscle diseases. Management must however be undertaken very early in these patients because outcome can be rapidly fatal after the development of significant desaturation. Nasal ventilation can improve not only quality of life but also life expectancy for these patients. PMID- 11924032 TI - Role of serotonin in sleep mechanisms. AB - Since near the time of its discovery over 40 years ago, the serotonergic system has been implicated in the regulation of the sleep-wake cycle. While early studies indicate that serotonin (5-HT) was associated with the initiation and maintenance of sleep, later studies indicate that serotonergic neurons also play a role in inhibiting sleep. As reviewed in this paper, the complex effects of 5 HT in the regulation of sleep is due in part to the fact that 5-HT can act at different areas of the brain that have been associated with the control of sleep and wake. In addition, the recent discovery of multiple 5-HT receptors through the mammalian brain led to the finding that different 5-HT receptors are selectively involved in the regulation of the different sleep states. PMID- 11924031 TI - [Pathology of craniocervical junction and sleep disorders]. AB - Sleep apnea is a manifestation which has recently been recognized in anomalies of craniovertebral junction. The main reported cases involve Arnold-Chiari malformation, and clinical manifestation is a central sleep apnea syndrome. The pathophysiological mechanisms of such a sleep apnea are a blunted bulbar chemical drive (in hypercapnic patients) or an increased bulbar chemical drive which destabilizes the breathing pattern during sleep (in normo/hypocapnic patients). As this sleep apnea can be the initial manifestation of a Arnold-Chiari malformation, craniocervical magnetic resonance imaging is recommended when a central sleep apnea without evident aetiology is discovered in a young patient. Other anomalies, such as bone malformations (platybasia for example) or syryngobulbomyelia can be involved. Other respiratory manifestations can be observed such as obstructive sleep apnea syndrome, diaphragmatic or vocal cord paralysis. These manifestations may be life-threatening and may lead to decompressive surgery. PMID- 11924033 TI - [Influence of cerebral and peripheral nitric oxide on sleep/wake cycle in the rat]. AB - Through a polygraphic and pharmacological approach conducted in the rat, it is established that L-NAME (N-nitro-L-arginine methyl ester; an inhibitor of the endothelial and neuronal NO-synthases), administered intraperitoneally (i.p., 100 mg/kg) increases significantly slow wave sleep (SWS) and paradoxical sleep (PS). On the contrary, 7-NI (7-Nitro-indazole; an inhibitor of the neuronal NO synthase), when administered locally in the nRD (n. raphe dorsalis), decreases PS without SWS changes. Finally, SIN-1 (molsidomine, a NO donor), also administered locally in the nRD (200 ng/0.2 microliter), increases PS without effects on SWS. According to the fact that L-NAME is not efficient on the cerebral NO fraction when administered i.p., it appears likely that this compound, mainly through its vascular effects, might inhibit SWS and PS. On the contrary, the effects, respectively inhibitory and facilitatory, observed on PS after nRD local injection of 7-NI and SIN-1 1 indicate that the NO-ergic component of this nucleus might facilitate PS. PMID- 11924034 TI - [Variations of hypothalamic and cortical prostaglandins and monoamines reveal transitions in arousal states: microdialysis study in the rat]. AB - Brain microdialysis coupled with EEG recording allowed us to track dynamic neurochemical changes every 3 or 6 minutes in relation to sleep/wake cycles. We chose to investigate prostaglandins (PG) and monoamines (catecholamines, serotonin and metabolites) because of their respective role in the states of vigilance, mainly suggested by pharmacological approaches, and because of the known interactions between PGs and monoamines. We focused on the paraventricular and ventromedial nuclei of the hypothalamus for their involvement in the relationships between feeding, metabolic rate and sleep, and the prefrontal cortex for its role in vigilance. These studies revealed a few changes in prostaglandin or monoamine levels as a function of a given state of vigilance. In particular, serotonin levels were higher during wakefulness than during sleep in both hypothalamus and cortex. Both hypothalamic and cortical PGE2 levels were higher during wakefulness than during slow wave sleep and still higher during paradoxical sleep in the cortex. Cortical PGD2 showed an exactly reverse profile of PGE2. These changes are in agreement with the described awaking action of PGE2 and with the hypnogenic action of PGD2. Our most informative findings were the sequential changes around transitions from one state to another that allow to predict the moment of onset of both sleep and wakefulness. Both in hypothalamus and in cortex, ondulatory patterns of PGE2 were encountered around the transitions between states. PGE2 was high in the middle of wakefulness, then regularly dropped announcing the occurrence of sleep, where the drop persisted before giving place to a rise in prediction to the next period of wakefulness. A similar profile was also observed for cortical serotonin, but its low levels reached a plateau during sleep. Cortical dopamine levels showed sudden and dramatic drops during short periods of wakefulness closely surrounding slow wave sleep. In some instances, as in the case of PGE2, similar profiles of variations could be found both in the hypothalamus and cortex. But in most cases, different and even opposite profiles were encountered in those two structures. Interestingly, in some instances, the pattern of changes in both prostaglandins and monoamines were similar, as for example between hypothalamic PGE2 and dopamine as well as between cortical PGE2 and serotonin. These similarities support the idea of the suggested interaction between prostaglandins and monoamines, in particular concerning their involvement in the regulation of sleep/wake cycles. PMID- 11924035 TI - [Vascular disorders and obstructive sleep apnea syndrome]. AB - The relationships between obstructive sleep apnoea syndrome (OSAS) and vascular diseases are still under discussion, but increasing evidence demonstrates that the OSAS is an independent risk factor for stroke, coronary artery diseases and hypertension. Many recent studies have found a 70 to 95 percent frequency of OSAS in patients with acute stroke. Furthermore the presence of OSAS in stroke patients could lead to a poor outcome. The potential mechanisms linking OSAS and stroke and probably multiple (arterial hypertension, cardiac arrhythmia, increased atherogenesis, coagulation disorders, and modifications of cerebral metabolism and hemodynamic). Despite numerous incertainties, OSAS should be systematically screened when it is clinically suspected in patients with acute vascular event (stroke, coronary artery diseases). However, the best time for treatment with continuous positive airway pressure remains to be determined. PMID- 11924036 TI - [Obstructive sleep apnea syndrome and heart failure]. AB - Heart failure has an increasing prevalence in middle age adults. The prognosis is very poor even with improved medical therapy and heart transplants. The outcome is related to the neurohumoral disease resulting from heart failure which leads to sympathetic activation that in turns worsens the prognosis. About half of the patients have sleep breathing disorders with variable proportions of central and obstructive apneas. Obstructive apneas are acutely deleterious to ventricular function. On the long run, they may be responsible for a worsening of the disease due to the permanent sympathetic activation seen in obstructive sleep apnea. It is therefore important to detect sleep apnea in patients and to apply a treatment. The best therapeutic procedure in obstructive events appears to be CPAP, provided hemodynamic status is closely monitored. PMID- 11924037 TI - [Obstructive sleep apnea syndrome and cardiovascular risk]. AB - Numerous studies have shown that obstructive sleep apnea syndrome (OSAS) is associated with an increased cardiovascular morbidity and mortality. Obstructive sleep apnea syndrome and cardiovascular disorders are frequent diseases. They share several risk factors such as age, gender, obesity, smoking, and alcohol. Therefore it is difficult to demonstrate that OSAS is a cardiovascular risk factor, independent of previously known factors. Recent epidemiological studies and trials, consistent with the results of previous studies, have demonstrated a strong association between OSAS and systemic hypertension. They also suggest that there is a possible cause-effect relation between OSAS and coronary artery disease or cardiac arrhythmias. Studies demonstrating that early recognition and treatment of OSAS may be effective in reducing these cardiovascular complications are still needed. PMID- 11924038 TI - Sleep and stroke. AB - Although the incidence of strokes is not maximal during sleeping hours, several lines of evidence make it probable that sleep in combination with breathing disorders like snoring and obstructive apneas are risk factors for ischemic strokes: the natural history of snoring and obstructive sleep apnea shows a higher incidence of strokes than in undisturbed sleep, the prevalence of snoring and sleep apneas in stroke patients is by far higher than in non-stroke patients; odds-ratios of stroke are higher in snorers and apneic patients than in normals, although the correction for confounders seems never perfect. The analysis of potential pathomechanisms linking sleep disordered breathing to strokes is another approach to the main topic: snoring and sleep apnea induce hypertension and arrhythmia, the carotid intima-media-thickness is increased, carotid atheromas are more common among apneics than among normals, the flow in the A. cerebri media is as well altered as the reaction to angiotensine II, noradrensine, isoproterenol and bradykinin. Homocysteine is increased, plasminogen activator inhibitor type 1 is inhibited and platelets are activated leading to an increased risk of thrombosis. There are no studies showing the effectiveness of treatment with nasal continuous positive airway pressure (nCPAP) on the rehabilitation of apneic stroke patients, but the outcome of non-apneic stroke patients is better than that of apneic stroke patients. PMID- 11924039 TI - [Is a genetic approach to insomnia possible?]. AB - The genetic approach of the sleep-wake cycle allowed the identification of several genes involved in the paradoxical sleep regulation and in narcolepsy. Concerning insomnia, a genetic study is more complex. Insomnia is indeed a wakefulness disorder defined by non quantitative biological parameters. The genetic approach of insomnia needs a more precise definition. PMID- 11924040 TI - Neuroendocrine dysregulation in primary insomnia. AB - Recent research has pointed to a functional link between stress, disturbed sleep, psychiatric disorders, ageing, and neuroendocrine dysfunctions. In particular, increased activation of the hypothalamic-pituitary-adrenal (HPA) axis--expressed as elevated plasma cortisol levels--was shown in physiological ageing and patients with psychiatric disorders. We found increased evening and nocturnal plasma cortisol concentrations in patients with primary insomnia. Considering that both ageing and psychiatric disorders are commonly associated with sleep disturbances, our results implicate that elevated cortisol concentrations are a rather unspecific feature of disturbed sleep. Furthermore, our data revealed a strong positive correlation between evening cortisol secretion and the number of nocturnal awakenings in both insomniac patients and controls. Since nocturnal exposure to increased HPA activity promotes sleep fragmentation even in healthy controls, increased evening cortisol levels may be a crucial factor in inducing and maintaining sleep disturbances. We therefore propose a model of HPA dysregulation in insomnia. This model is based on the arousal theory of insomnia and the strong correlation between evening cortisol secretion and sleep fragmentation as a pathophysiological mechanism of a vicious cycle of insomnia. In patients with long-lasting insomniac complaints we found decreased nocturnal plasma melatonin levels thereby indicating a labilisation of circadian rhythm functions. Taken together, the neuroendocrine dysregulation seems to be more expressed in chronic insomnia than in acute insomnia and may be a contributing factor in maintaining disturbed sleep. PMID- 11924041 TI - [Neurophysiological basis of insomnia: role of cyclic alternating patterns]. AB - During non-REM (NREM) sleep it is possible to identify two complementary conditions of arousal stability and arousal instability. Unstable sleep, which can be detected in all stages, is expressed by the recurrence of arousal complexes (sequences of K-complexes, delta bursts, K-alpha, conventional arousals), which translate a brief (10-15 s) activation of the sleeping brain. These repetitive arousal complexes compose the cyclic alternating pattern (CAP). During stable NREM sleep, arousal complexes are rare or absent and the EEG lacks any cyclic pattern (non-CAP). CAP is a spontaneous feature of normal sleep as it is typically involved in stage changes and nocturnal motor activity. The physiological amount of CAP rate (ratio of CAP time to NREM sleep time) varies with age according to a U-shape curve. Within these ranges, sleep is perceived as continuous and restorative. Conversely, an excess of CAP rate fragments sleep and impairs its quality. Polysomnographic investigation reveals that untreated insomniac patients exhibit significantly higher values of CAP rate compared to healthy sleepers. Effective hypnotic treatment restores physiological amounts of CAP rate, with specific differences between the administered drugs. The sensitivity of CAP parameters to drug manipulation can provide circumscribed information on the hypnotic properties of active compounds. PMID- 11924042 TI - [Psychological vulnerability to insomnia]. AB - Insomnia in widespread health with important personal, occupational, and economic consequences. Multiple causes (psychologic, medical, pharmacologic, environmental) are recognized, but little is known about risks predisposing certain individuals to chronic insomnia. For instance, what is the role of psychological vulnerability? Although the response to this question might appear obvious, valid scientific evidence is lacking. We present a conceptual model of insomnia distinguishing vulnerability, triggering and maintaining factors. We also discuss different types of evidence supporting the notion of psychological vulnerability in insomnia. Finally, we examine the clinical implications and propose avenues of future research. PMID- 11924043 TI - [Adenosine in sleep regulation]. AB - Sleepiness increases with duration of sleep deprivation. Rebound sleep together with enhancement of slow wave activity are characteristic of the subsequent recovery period. These homeostatic properties of the regulation of sleep wakefulness are mediated by central adenosinergic modulations. The involvement of adenosine in sleep processes has been known for a long time, as illustrated by the fact that blockade of adenosine receptors by caffeine promotes wakefulness. However, its mechanisms of action in these processes have only recently been thoroughly investigated, notably by use of microdialysis techniques in free moving animals. In the central nervous system, adenosine acts as a neurotransmitter, but it is not released from synaptic vesicles in specific neurons. Adenosine is synthesized in neurons and glial cells and is released into the extracellular space when energy expenditure exceeds energy production. Adenosine acts at A1 receptors to inhibit target transmitter release and to hyperpolarize neurons, and at A2A receptors, possibly activating GABAergic inhibitory systems. Extracellular concentrations of adenosine in most brain areas are deceased during sleep compared to wakefulness, but these modifications are linked to sleep regulatory mechanisms, essentially in the basal forebrain. In the basal forebrain and in the cortex, adenosine levels are considerably enhanced during sustained prolonged wakefulness and decrease progressively during the recovery period. This enhancement of adenosine concentration would be responsible for sleep rebound and for slow wave activity observed after sleep deprivation, since these effects are mimicked by infusion of A1 agonists into the basal forebrain and are counteracted by treatment with adenosinergic antagonists such as caffeine or theophylline. The action of adenosine on sleep-wakefulness regulation would be accounted for by an inhibitory influence on wakefulness promoting cholinergic neurons of the basal forebrain and the mesopontine area, and by facilitation of sleep-related neurons in the hypothalamic preoptic nucleus. Through these mechanisms, adenosine would be, more than a sleep modular, a homeostatic signal regulating sleepiness and sleep rebound, which are both associated with prolonged wakefulness. PMID- 11924044 TI - [Motor control of the upper airways]. AB - During sleep upper airway resistance increases proportionally with a slightly diminished air flow. This can however be compensated for by increasing inspiratory force that further aggravates airway collapse and resistance. Apnea during sleep results from an interaction between the different respiratory and pharynx muscles. Sleep modifies the activity of these muscles leading to increased resistance and muscle fatigue, then eventually histological modifications. Electrical activity is not always proportional to the generated force and muscle shortening, so the effect is not always an increased permeability. In apneic patients, the efficacy of these muscles is preserved but tissue response may be poor. Histological adaptations are variable and depend on the level of the pharynx and the variability of the site of obstruction. This raises the question of electrical stimulation of certain muscles and the choice of the site of stimulation. PMID- 11924045 TI - [Obstructive sleep apnea syndrome and the upper airway muscles]. AB - Pharyngeal muscles are prone to distorsion during inspiratory negative pressure. Some pharyngeal muscles, called pharyngeal dilatators, exhibit tonic and/or phasic inspiratory activity. At the velar level, tensor palatini, glossopharyngeus, palatopharyngeus and musculus uvula drive the airflow towards nasal or buccal breathing. Genioglossus and geniohyoid muscles exert a forward propulsion to the tongue. The contraction of the dilatators muscles precedes the diaphram contraction. These muscles show a poor endurance. In obstructive sleep apnea syndrome, they contract during unfavorable metabolic (hypoxia and hypercapnia) as well as mechanical (excentrical contraction) conditions. Histological changes occur in upper airway dilator muscles: muscle volume and proportion of type Ila fibers are increased. These changes, considered as compensatory mechanisms, are variable within the different studied pharyngeal muscles. The initial pharyngeal obstructive site, and its further extension, could be determined by the different strength and endurance properties of these muscles. PMID- 11924046 TI - [Imaging in obstructive sleep apnea syndrome]. AB - Obstructive sleep apneas result from pharyngeal collapse during sleep. Upper airways may be investigated using either static or dynamic techniques during wakefulness or sleep. In OSAS patients, imaging has different goals: to describe the anatomy of the pharynx and the sequence of the pharyngeal collapse, to try to select patients that will benefit from surgery, and to analyze the results of different treatments. PMID- 11924047 TI - Nocturnal sleep features in narcolepsy: a model-based approach. AB - The internal structure of night sleep in subjects with narcolepsy significantly differs as compared to the controls. The differences consist in the presence of sleep onset REM periods (SOREMPs) and in a longer duration (120 minutes versus 90 minutes) of the NREM-REM cycles. Another difference consists in the lack of increase in REM sleep duration during the night, cycle after cycle. These characteristics, taken as a whole, suggest that the sleep structure of narcoleptic patients is due to an imbalance between the homeostatic process and the enhanced pressure of REM inducing mechanisms. With regard to the intrasleep dynamics the current extension of the Two-process model cannot give a satisfactory explanation because REM sleep is included as an external trigger with no intrinsic rhythmic property. Neurobiologic studies lead to consider the REM cyclic occurrence as the result of reciprocal interaction between two populations of REM-ON and REM-OFF cells. The introduction in the Two-process model extended to the intra-night dynamics of an ultradian oscillator, based on a REM-ON--REM-OFF reciprocal interaction, allows the theoretical possibility to simulate a SOREMP. This revised model accounts for the progressive extension of REM sleep-periods duration in the course of the night. The sleep structure of narcoleptic patients can be simulated leaving unaltered the pressure of the homeostatic process and enhancing the REM inducing pressure. Such an enhanced REM pressure can be achieved by increasing the value of the numerical coefficient, which represents the strength of connection between the two types of REM-ON and REM-OFF cells. This modification allows not only to obtain a stronger ultradian oscillator but also a longer periodicity of REM sleep occurrence. By coupling the homeostatic process, considered as normal, to such a modified ultradian oscillator, our model can explain the intra-night sleep dynamics of narcoleptic subjects. A REM-ON REM-OFF dysregulation can be hypothesized to explain the pathophysiological basis of nocturnal sleep features in narcolepsy. PMID- 11924049 TI - [Genetics of human narcolepsy]. AB - Narcolepsy is known to be a complex disorder; both genetic and environmental factors play a role in its pathophysiology. Although narcolepsy presents one of the tightest association with a specific HLA antigen (DQB1*0602), there is strong evidence that non-HLA genes also confer susceptibility, both monoaminergic and hypocretinergic systems seem to be involved and may interfere with the phenotype. Implication of the hypocretin system is well-established in both canine and murine narcolepsy (caused by mutation) and a consistent reduction in hypocretin neuron seems to be the cause of human narcolepsy. An autoimmune process is probable. PMID- 11924048 TI - [Circadian and ultradian cycles in narcolepsy]. AB - In narcolepsy, homeostatic process is preserved while sleep/wake circadian process is impaired. Other circadian components (body temperature, endocrine secretions, subjective sleepiness) are preserved. This circadian system weakness permits the occurrence of a very strong ultradian component, modulating sleep/wake rhythm. So, a 4 hour ultradian rhythmicity of Slow Wave Activity, and a 2 hour NREM/REM cycle longer than in normal subjects, has been evidenced in narcoleptic patients. These circadian and ultradian alterations can explain a major part of the narcoleptic symptoms. PMID- 11924050 TI - [Immunological aspects of narcolepsy]. AB - Since the discovery of an almost 100 p. cent association of human narcolepsy with the HLA gene DQB1*0602, research has been focused on autoimmune mechanisms. Epidemiological data (age of onset, triggering factors, association with multiple sclerosis) would lend support to this hypothesis. However it has remained largely impossible to demonstrate immune abnormalities in blood or CSF by means of usual techniques. The canine form of the disease was supposed to be also immunologically mediated, since a linkage with a human immunoglobulin-related gene had been demonstrated. This was eventually demonstrated to be a pseudo linkage, the real cause being a mutation in the closely related hypocretin receptor gene. This recently discovered neuropeptide is clearly involved in some aspects of sleep regulation. Soon thereafter, hypocretin deficiency was found in human narcoleptics, due to a severe neuronal loss in the hypothalamus; gliosis having been evidenced, it may be considered as the evidence of a prior inflammatory reaction, possibly due to an immune attack. PMID- 11924051 TI - "Net-notes for nurses". PMID- 11924052 TI - Professional liability insurance. Myth vs. fact. PMID- 11924053 TI - Is your employer's coverage enough? PMID- 11924054 TI - Final congressional action: House, Senate pass Nurse Reinvestment Act; VA nurse bill cleared for president's signature. PMID- 11924055 TI - Coalitions of nursing specialty organizations unite. PMID- 11924056 TI - If I could do it all over again, I would become a nurse. PMID- 11924057 TI - The diabetes epidemic doesn't have to be part of African American history. PMID- 11924058 TI - [Briefly stated]. PMID- 11924059 TI - [Do we have too many or too few nursing personnel?]. PMID- 11924060 TI - [Technical anesthesia assistants--yes or no?]. PMID- 11924061 TI - [Guidelines for resuscitation of circulation and respiration]. PMID- 11924062 TI - [Risks and benefits. Performance assessment in intensive care]. PMID- 11924063 TI - [Who is responsible for malpractice in the operating room? 1: Tutoring in malpractice law]. PMID- 11924064 TI - [Personnel development in geriatric nursing. 1]. PMID- 11924065 TI - [Preventing the risks of accidental falls. The Hendrich Accidental Falls Risk Scale]. PMID- 11924066 TI - Quality professionals must take lead to eliminate wrong-site surgery. PMID- 11924067 TI - Pain management is high priority for surveyors. PMID- 11924068 TI - Patient safety: just old wine in a new bottle? PMID- 11924069 TI - Bioterrorism watch. Ease of access to deadly chemicals may be the greatest threat to hospital readiness. PMID- 11924071 TI - Bioterrorism watch. Signs and symptoms of chemical exposures. PMID- 11924072 TI - Hypokalemic thyrotoxic periodic paralysis: a case report and review. AB - Hypokalemic thyrotoxic periodic paralysis (HTPP) is a rare disorder affecting primarily men of Asian descent. Males are 20 times more likely to be affected than females. The following case describes a young Native American female with HTPP. PMID- 11924073 TI - Make colorectal cancer screening. A part of routine care. PMID- 11924074 TI - Universal leukocyte reduction--is it worth it? PMID- 11924075 TI - Anticonvulsant-induced bone disease. PMID- 11924076 TI - A world without sanctuary. PMID- 11924077 TI - Generating hypotheses after 14 years of marital followup; or, how should one speculate? A reply to DeKay, Greeno, and Houck. AB - Gottman and Levenson (2002) for the purpose of post hoc speculation, developed a number of ordinary least squares regressions to model the length of marriage of divorcing couples in a 14-year longitudinal study. We believe that our analyses are appropriate for our purpose. We do not agree with DeKay, Greeno, and Houck (2002) that a duration-model approach would have been more appropriate, and instead argue that the analyses used are more powerful and generate more interesting speculations. When speculating, one makes an important contribution just by being interesting, not necessarily by being right. The purpose of post hoc speculation is to generate discussion, and we are pleased that even at the outset we have accomplished this goal. In this reply to DeKay et al., we argue that the two-process model for earlier versus later divorcing that we propose is both interesting and clinically useful. PMID- 11924078 TI - Blinding and deafening moments, and threatening futures: in the wake of September 11, 2001. PMID- 11924079 TI - Counseling and couple therapy for infertile couples. AB - The article describes a two-tier, interdisciplinary design for the psychological counseling and therapy of childless couples. It is solution- and resource oriented and avoids psychopathological ascriptions. Couples are supported in coming to terms with the crisis of a physical disorder and its emotional consequences; they are also aided in developing prospects and options for a future without a biological child. The procedure is explained in detail and provides a model suitable for application at reproduction medicine centers and gynecological and andrological practices. Sample interventions illustrate the therapeutic attitude advocated. PMID- 11924080 TI - Clinicians' fidelity to a manual-based family treatment as a predictor of the one year course of bipolar disorder. AB - This study assessed whether therapist adherence to the family focused treatment model for patients with bipolar disorder and their relatives was associated with patient outcomes at one year after treatment entry. A total of 78 videotaped sessions of FFT consisting of 26 families with a member with bipolar disorder (3 sessions/family) were rated on fidelity using the Therapist Competence/Adherence Scale (TCAS; see Endnote 1, p. 130). Patients' outcomes (relapse status) were assessed using the Brief Psychiatric Rating Scale (BPRS) and selected items from the Schedule of Affective Disorders and the Schizophrenia-Change (SADS-C) scale (measured at 3-month intervals for 12 months). Contrary to expectations, therapist fidelity was not related to overall outcome as assessed by the BPRS and the SADS-C. Among patients who did relapse, higher levels of cooperation among therapists predicted a later date for relapse than did lower levels of cooperation. Surprisingly, and in opposition to the study's hypotheses, patients who were hospitalized because of relapses had therapists who were rated as more competent in their ability to conduct the problem-solving module of FFT. Study implications are discussed. PMID- 11924081 TI - Ambiguous loss: working with families of the missing. PMID- 11924082 TI - The power of community. PMID- 11924083 TI - The helpers and the helped: viewing the mental health profession through the lens of September 11. PMID- 11924084 TI - Living with threat and uncertainty: what the Kosavars tell us. PMID- 11924085 TI - The loss of innocence. PMID- 11924086 TI - Why do they hate us? PMID- 11924087 TI - Hard times for the forces of compassion. PMID- 11924088 TI - Bouncing forward: resilience in the aftermath of September 11. PMID- 11924089 TI - Healing traumatized children: creating illustrated storybooks in family therapy. AB - In this article we describe the therapeutic practice of creating illustrated storybooks in family therapy with traumatized children. Illustrated stories offer a predictable structure to sessions and facilitate engagement and participation of children in therapy. The therapeutic emphasis of storybooks can be adjusted to take into account a child's life story, verbal capacity, level of anxiety, and traumatic hyperarousal. The creation of storybooks is an active process that embraces important aspects of trauma-specific interventions, including expression of trauma-related feelings; clarification of erroneous beliefs about the self, others, or the traumatic event; and externalization of traumatic stimuli into artwork, allowing for exposure and habituation of the arousal response. A focus on visual images together with narrative takes advantage of children's developmental capacities and spontaneous pleasure in the creation of art, thus minimizing anxiety and enhancing feelings of mastery, competence, and hope. The creation of storybooks is compatible with family interventions that foster a safe family context, strengthen attachment relationships, insure appropriate structure and boundaries, and enhance parenting capacity as well as those interactions that facilitate understanding and dialogue between family members. PMID- 11924090 TI - Conversations with Salma Abugideiri: to live the greater jihad. PMID- 11924091 TI - Notes for a cultural history of family therapy. AB - The official history of family therapy describes its beginnings as a daring technical and philosophical departure from traditional individual treatment in the 1960s, inspired especially by the "system thinking" of Gregory Bateson. This celebrated origin story needs to be supplemented with a longer and larger history of both practice and thought about the family, and that is the subject of this article. The longer history goes back to the founding of social work by Mary Richmond, of pragmatism by William James, and of the organic view of social systems intervention by John Dewey. Seen against this background, family therapy is, among other things, a consequence of the development of persistent elements of American professional culture, experience, and philosophy. The taking of this historical-anthropological view discloses also the origins of two other histories that have made their contribution to the development of family therapy: a science of observing communication processes that starts with Edward Sapir and leads to contemporary conversation analysis, and a history of mesmerism in the United States that culminates in Milton Erickson and his followers. PMID- 11924092 TI - A two-factor model for predicting when a couple will divorce: exploratory analyses using 14-year longitudinal data. AB - This article examines 14-year longitudinal data and attempts to create a post hoc model that uses Time-1 data to "predict" the length of time the marriage will last. The sample consists of the 21 couples (of 79 studied) who divorced over a 14-year period. A two-factor model is proposed. One factor is the amount of unregulated volatile positive and negative affect in the marriage, and this factor predicts a short marriage length for the divorcing couples. A second factor is called "neutral affective style," and this factor predicts a long marriage length for the divorcing couples. This model is compared to a Time-1 model of ailing marriage in which Time-1 marital satisfaction is used to predict the timing of divorce. PMID- 11924093 TI - As a Japanese American. PMID- 11924094 TI - Searching for a two-factor model of marriage duration: commentary on Gottman and Levenson. AB - Gottman and Levenson (2002) report a number of post hoc ordinary least squares regressions to "predict" the length of marriage, given that divorce has occurred. We argue that the type of statistical model they use is inappropriate for answering clinically relevant questions about the causes and timing of divorce, and present several reasons why an alternative family of models called duration models would be more appropriate. The distribution of marriage length is not bimodal, as Gottman and Levenson suggest, and their search for a two-factor model for explaining marriage length is misguided. Their regression models omit many variables known to affect marriage length, and instead use variables that were pre-screened for their predictive ability. Their final model is based on data for only 15 cases, including one unusual case that has undue influence on the results. For these and other technical reasons presented in the text, we believe that Gottman and Levenson's results are not replicable, and that they should not be used to guide interventions for couples in clinical settings. PMID- 11924095 TI - The notion of heredity was first known in Islamic thinking some 1400 years ago. PMID- 11924096 TI - Genetic diseases in Arab populations. PMID- 11924097 TI - Spectrum of genetic disorders and the impact on health care delivery: an introduction. AB - Until recently, infectious diseases and malnutrition-related disorders constituted the major cause of ill health and mortality in the world population. However, advances in treatment of such disorders and increased understanding of the molecular basis of heredity have led to genetically transmitted conditions becoming a major cause of morbidity and mortality. Several disorders, including chromosomal (Down syndrome, Turner syndrome), single-gene (sickle-cell disease, thalassaemia, glucose-6-phosphate dehydrogenase deficiency, haemophilia, inborn errors of metabolism) and multifactorial disorders (coronary artery disease, arteriosclerosis, diabetes mellitus, hypertension, obesity) are common and becoming increasingly important. As there is no agreed-upon definitive cure with acceptable risk, these disorders are a significant burden on the health care delivery system. This is because the chronic nature of genetic diseases requires lifelong medical attention, expensive supportive and symptomatic therapy and specialist care. This review outlines the genetic disorders, their impact on health care delivery systems and the general framework required to prevent and control these disorders. PMID- 11924098 TI - Review of the spectrum of genetic diseases in Bahrain. AB - This paper looks at some of the studies on genetic disorders conducted in Bahrain. The disorders covered include: genetic blood disorders, metabolic disorders, chromosomal disorders, including Down syndrome, and cystic fibrosis. The rate of consanguinity in Bahrain and the results of premarital counselling are also discussed. PMID- 11924099 TI - Preventive aspects of genetic morbidity: experiences of the Canadian model. AB - Genetic disorders are emerging as a major health problem in industrialized countries, especially with greater control of environmental diseases. Of total admissions to major paediatric hospitals, 50% are due to genetic disorders, and at least 10% of all adult admissions to hospitals involve major genetic contributions. Several programmes aimed at preventing or ameliorating these genetic disorders are being implemented. Identifying people at risk of genetic disease has helped decrease the burden of such diseases on families and society. Early recognition also leads to greater success of treatment and improves outcome and prognosis. In Arab populations, genetic disorders are still not perceived as a major health problem, even though they are widely prevalent. Applying similar programmes of early detection, maternal screening, neonatal screening, carrier testing and susceptibility testing will significantly help reduce the impact of these disorders in our populations. PMID- 11924100 TI - Counselling about genetic disease: an Islamic perspective. AB - Genetic counselling is the process by which an individual or a family obtains information about a genetic condition that may affect them, so that they can take the appropriate decisions about marriage, reproduction and health management. Islamic teaching encourages counselling and stresses that the counsellor should be considerate, compassionate and should keep the secrets of the person or family involved. He/she should not impose his/her views on the clients. Some Arab countries encourage premarital medical examination to detect carriers of common hereditary diseases such as thalassaemia. However, this is still controversial, as it infringes the human rights of individuals. Reproductive options open to carriers of hereditary diseases are outlined in this paper, such as prenatal diagnosis, adoption, donation of a sperm, ova or preembryo and preimplantation diagnosis, and their acceptability within Islam is discussed. PMID- 11924102 TI - Human mutations and their detection by gene and linkage analysis, allele sharing and association methods. AB - It has been 20 years since DNA analysis was first used in the detection of sickle cell anaemia. Here, techniques for detecting human mutations are reviewed. We describe direct detection of mutations using restriction enzyme analysis and polymerase chain reaction amplification to detect gene deletions, rearrangements and point mutations. Indirect detection of mutations include the use of DNA polymorphisms in linkage analysis. PMID- 11924103 TI - Appraisal of sickle-cell and thalassaemia genes in Saudi Arabia. AB - A comprehensive national survey of the distribution of the sickle-cell (Hb S) gene and thalassaemia genes was initiated in 1982, with more than 30,055 blood samples collected. The Hb S, alpha- and beta-thalassaemia gene frequency range was 0.005-0.145, 0.01-0.40 and 0.01-0.15 respectively in various areas of Saudi Arabia. We present here an appraisal of sickle-cell and thalassaemia gene occurrence in the Saudi population, based on our studies conducted over 10 years in different regions of Saudi Arabia. PMID- 11924101 TI - Potential usefulness of preimplantation genetic diagnosis in the control and prevention of genetic diseases. AB - Prenatal diagnosis of molecular mutations can be of immense value, since diagnosis followed by genetic counselling provides the most appropriate approach to genetic diseases control and prevention. However, ethical, psychosocial and religious considerations hamper adoption of prenatal diagnosis in communities where termination of a pregnancy may not be acceptable. Recently, preimplantation genetic diagnosis has attracted considerable interest. This involves in vitro fertilization, followed by genetic disorder diagnosis using polar bodies or cells extracted from a blastomere stage. The normal blastomere is implanted in the womb and pregnancy proceeds naturally. If an abnormality is diagnosed, the blastomere is not implanted, thus preventing pregnancy with the affected fetus. This paper outlines the potential usefulness of preimplantation genetic diagnosis in the control and prevention of genetic disease in our part of the world. PMID- 11924104 TI - Haplotypes of the beta-globin gene as prognostic factors in sickle-cell disease. AB - We collaborated with researchers from Egypt, Syrian Arab Republic and Jordan in a study of patients with sickle-cell disease from those countries, and from various parts of Saudi Arabia, in order to investigate the influence of genetics on the clinical presentation of the disease, and to attempt to determine the origin of the sickle-cell gene in Arabs. Our results suggest that beta-globin gene haplotypes influence the clinical presentation of sickle-cell disease, and that there are at least two major foci for the origin of the sickle-cell gene, one in the eastern part of Saudi Arabia, and the other in the populations of North Africa and the north-western part of the Arabian peninsula. PMID- 11924105 TI - Pattern for alpha-thalassaemia in Yemeni sickle-cell-disease patients. AB - A group of Yemeni patients with sickle-cell disease (SCD) and normal Hb AA individuals living in Riyadh were studied to determine the incidence of the alpha gene molecular defect. Blood samples were obtained from 26 SCD patients and 19 controls (the Hb AA group). In the SCD patients the frequency of single alpha gene deletion (-alpha/alpha alpha) was 0.346, compared to 0.263 in the Hb AA group. The frequency of two gene deletion (-alpha/-alpha) was 0.231 (0.0 for the Hb AA group). In one Hb AA case, a triple alpha-gene arrangement (alpha alpha alpha/alpha alpha) was found (frequency 0.053). The results suggest that alpha thalassaemia occurs frequently in Yemeni SCD patients. Further studies to determine the overall frequency of alpha-thalassaemia in the Republic of Yemen would be of value for patient management. PMID- 11924106 TI - Prevalence of thalassaemia in schoolchildren in north-eastern Badia, Jordan. AB - The prevalence of the different types of thalassaemia and that of iron deficiency anaemia was investigated in 1020 schoolchildren (age range: 6-15 years) from the north-eastern Badia region of Jordan. beta-thalassaemia minor was the most prevalent (3.04%), followed by alpha 2-thalassaemia (2.06%). Only three cases of alpha 1-thalassaemia and one case of beta-thalassaemia major were found. Iron deficiency anaemia was diagnosed in 54 children (5.3%) (33 males, 21 females). The mean values of the blood characteristics of the normal, haemoglobinopathic and iron-deficient children were examined and compared. PMID- 11924107 TI - Haemostatic disorders in nonsplenectomized and splenectomized thalassaemic children. AB - A group of 40 thalassaemic patients (20 splenectomized and 20 nonsplenectomized) from the Haematology Unit of Tanta University Hospital (age range: 3-14 years) were studied to identify the mechanisms by which haemorrhagic and thrombotic complications occur in thalassaemic patients. The patients' levels of protein C, antithrombin III and in vitro platelet aggregation in response to collagen were compared with those of 20 controls. The study suggests that thrombocytosis, increased platelet aggregation and decreased natural coagulation inhibitors (protein C and antithrombin III) in splenectomized thalassaemic children may be significant in thrombotic complications in such patients. Defective platelet aggregation and prothrombin activity in nonsplenectomized children may also give rise to haemorrhagic tendencies. PMID- 11924108 TI - Communicating basic genetics to patients with sickle-cell disease. AB - This report describes an approach to explaining the basic genetics of sickle-cell disease to patients and their families in an uncomplicated yet sufficiently informative way. The areas covered include: the cause of the disease, its pathophysiology, and the importance of genetic counselling. PMID- 11924109 TI - Molecular studies on Yemeni sickle-cell-disease patients: Xmn I polymorphism. AB - Our studies of the Saudi population have shown that in patients with mild presentation of sickle-cell disease (SCD) from Saudi Arabia's eastern region, the prevalence of polymorphic sites is high. However, the prevalence is very low in patients with severe SCD from the south-west of the country. We expanded these studies to a group of Yemeni patients with severe SCD, resident in Riyadh. We investigated a total of 60 chromosomes carrying the sickle-cell (Hb S) gene and 14 chromosomes carrying the Hb A gene. Amongst the Hb AA group, the prevalence was 42.9% and 57.1% for the presence (+) and absence (-) of Xmn I polymorphic sites. In the Hb SS individuals, the prevalence of Xmn I polymorphic sites was similar to the prevalence reported in the south-western region of Saudi Arabia. PMID- 11924110 TI - Spectrum of hereditary coagulation factor deficiencies in eastern province, Saudi Arabia. AB - In a 7-year retrospective analysis (1991-97) of the records of the Department of Haematology, Regional Laboratory and Blood Bank (Dammam), 54 patients from all parts of Saudi Arabia's Eastern Province were diagnosed with hereditary coagulation factor deficiencies. The largest group of patients, 42 haemophiliacs, included 4 non-Saudi patients. There were 39 haemophilia A or factor VIII deficiency patients, 2 haemophilia B or factor IX deficiency patients and 1 combined factor VIII and V deficiency patient. There were 5 Saudi patients with probable factor XIII deficiency, and 7 patients, all but one who were Saudi, had von Willebrand disease. The distribution of haemophilia patients in Eastern Province showed some differences compared with those reported from Riyadh and from Western countries. Among Saudis in Eastern Province, the number with suspected factor XIII deficiency, although low, was higher than that reported for other regions. The number of patients with haemophilia B and von Willebrand disease was lower than expected, when compared with the number of haemophilia A cases. PMID- 11924111 TI - Classic homocystinuria: clinical, biochemical and radiological observations, and therapeutic outcome of 24 Saudi patients. AB - We considered the clinical, biochemical and radiological findings, and response to pyridoxine (vitamin B6) of 24 classic homocystinuric patients (15 females, 9 males) diagnosed at King Faisal Specialist Hospital and Research Centre. Common clinical findings included ectopia lentis (20 patients), skeletal system involvement (18 patients), vascular system involvement (9 patients) and mental retardation (all patients to varying degrees). A number of unusual findings were reported. The parents of 21 patients were first-degree relatives and 19 patients had at least one other family member affected by the same disease. Only 4 patients responded to pyridoxine; their methionine level decreased to almost normal range. PMID- 11924112 TI - Tyrosinaemia type II: an easily diagnosed metabolic disorder with a rewarding therapeutic response. AB - We retrospectively reviewed clinical and biochemical data of four patients diagnosed with tyrosinaemia type II. Diagnosis was established by high plasma tyrosine and normal plasma phenylalanine levels using plasma high-pressure liquid chromatography and tandem mass spectrometry. All patients were mildly mentally retarded and had painful non-pruritic and hyperkeratotic plaques on the soles and palms. There were no ophthalmic symptoms. The patients dramatically responded clinically and biochemically to a diet restricted in tyrosine and phenylalanine. PMID- 11924113 TI - Glutathione reductase deficiency in Saudi Arabia. AB - Glutathione reductase (GR) is a ubiquitous enzyme required for the conversion of oxidized glutathione (GSSG) to reduced glutathione (GSH) concomitantly oxidizing reduced nicotinamide adenine dinucleotide phosphate (NADPH) in a reaction essential for the stability and integrity of red cells. Mutations in the GR gene and nutritional deficiency of riboflavin, a co-factor required for the normal functioning of GR, can cause GR deficiency. We conducted a study on 1691 Saudi individuals to determine the overall frequency of GR deficiency and to identify whether the deficiency results from genetic or acquired causes or both. The activity of GR was measured in freshly prepared red cell haemolysate in the presence and absence of flavin adenine dinucleotide (FAD) and the activity coefficient (AC) was determined. Samples with low GR activity (> 2.0 IU/g haemoglobin) both in the presence and absence of FAD and an AC between 0.9 and 1.2 were considered GR-deficient. Samples with AC > or = 1.3 were considered riboflavin-deficient. The overall frequency of partial GR deficiency was 24.5% and 20.3% in males and females respectively. In addition, 17.8% of males and 22.4% of females suffered from GR deficiency due to riboflavin deficiency. This could be easily corrected by dietary supplementation with riboflavin. No cases of severe GR deficiency were identified. PMID- 11924114 TI - Biotinidase deficiency: a treatable genetic disorder in the Saudi population. AB - Biotinidase deficiency is an autosomal recessive genetic disorder which is not uncommon in the Saudi population. Biotinidase is responsible for biotin recycling and biotin is an essential cofactor for activation of the carboxylase enzymes. Absence of biotinidase leads to infantile or early childhood encephalopathy, seizure disorder, dermatitis, alopecia, neural deafness and optic atrophy. The disease can be diagnosed by simple fluorometric enzyme assay. Treatment with biotin is both cheap and simple, resulting in rewarding clinical recovery and normalization of the biochemical, neuroradiological and neurophysiological parameters. If neglected, however, a patient may die of acute metabolic acidosis or may suffer from permanent neural deafness and optic atrophy, with mental and motor handicap. We describe the detection and treatment of 20 cases of biotinidase deficiency in our hospital and recommend the introduction of a neonatal screening programme for this disorder. PMID- 11924115 TI - Use of FISH technique in the diagnosis of chromosomal syndromes. AB - Major chromosome abnormalities are present in 0.65% of all neonates. Fluorescent in situ hybridization (FISH) is useful in diagnosing microdeletion syndromes that would otherwise be difficult to diagnose using standard cytogenetics. In this study, we used FISH analysis in the laboratory diagnosis of 4 patients with Prader-Willi Syndrome [del(15)(q11.2q12)], 4 patients with DiGeorge syndrome [del(22)(q11.2q11.23)] and 4 patients with Williams syndrome [del(7)(q11.23q11.23)]. High-resolution chromosome analysis in all these patients was either normal or inconclusive but all the syndromes were confirmed using FISH. We recommend cytogenetic analysis should always be supplemented with FISH to diagnose all cases suspected of a microdeletion syndrome. PMID- 11924116 TI - Application of DNA-based tests for diagnosis of spinal muscular atrophy in Saudi Arabia. AB - We examined the deletion of the survival motor neuron (SMN) and neuronal apoptosis inhibitory protein (NAIP) genes in patients with spinal muscular atrophy (SMA) using polymerase chain reaction followed by restriction site assay methods. The study included 16 Saudi patients (9 SMA type I and 7 SMA type II) and 6 healthy Saudi volunteers. The homozygous deletions of exons 7 and 8 of the telomeric SMN gene, and exon 5 of the NAIP gene were found in all SMA type I patients. Exons 7 and 8 of telomeric SMN were deleted in all SMA type II patients. However, exon 5 of NAIP was deleted in three of the seven cases. All control volunteers and all family members of the patients had normal SMN and NAIP. The incidence of NAIP deletion was higher in the more severe SMA cases and the dual deletion of the SMN and NAIP genes was more common in Saudi SMA type I patients compared with patients of other ethnic groups. PMID- 11924117 TI - Geographic distribution of cystic fibrosis transmembrane regulator gene mutations in Saudi Arabia. AB - A descriptive study was undertaken to characterize cystic fibrosis transmembrane regulator (CFTR) gene mutations in the Saudi Arabian cystic fibrosis (CF) population in relation to their clinical picture, demographic features and ethnic origin. From October 1992 to September 1997, 70 patients (46 families) diagnosed with CF were screened for CFTR mutations. A total of 12 mutations were identified in 34 families (70% of the CF alleles in the study group). Most of the families were native Saudis, and in 88% of the families the parents were in consanguineous marriages. The most common Saudi mutations were 1548delG and I1234V. There was no significant difference in the clinical picture between patients of different ethnic origins with the same CFTR mutation. PMID- 11924118 TI - Diabetes mellitus, hypertension and obesity--common multifactorial disorders in Saudis. AB - Diabetes mellitus, hypertension and obesity are among the multifactorial disorders that occur at a higher prevalence in older age groups. Their prevalence is affected by both genetic and environmental factors. We investigated the distribution of diabetes mellitus, hypertension and obesity in Saudi males and females by conducting a household screening survey during the period 1992-1996 of the adult population (> 14 years) in five different areas of Saudi Arabia. Height, weight, age and other essential details were recorded and diastolic and systolic blood pressures measured. Glucose levels were measured in blood taken after fasting and 2 hours after a glucose load. The data were used to classify the individuals as diabetic, glucose intolerant and normal, using WHO criteria. The individuals were further classified as type 1 diabetes mellitus and type 2 diabetes mellitus. The overall prevalence of diabetes mellitus was 9.7% and 7.0%, obesity 13.05% and 20.26%, overweight 27.23% and 25.20%, and hypertension 5.39% and 3.65% in the adult male and female populations respectively. A significant increase was observed in the prevalence of diabetes, obesity and hypertension with age in both males and females. In addition, the prevalence of obesity and overweight was significantly higher in the individuals with diabetes mellitus. PMID- 11924119 TI - [Heterozygosity in patients with pulmonary tuberculosis showing varying responses to therapy]. AB - The distribution of the levels of heterozygosity was analyzed by 9 loci of genetic markers: PI, TF, PGM1, ACPI, HP, GC, GLO1 C3, and ESD in two groups of patients with pulmonary tuberculosis who had improvements (Group 1, n = 71) and failures (Group 2, n = 35). The heterozygosity observed in the groups was compared with that calculated by the Hardy-Weinberg law by using data on healthy controls (n = 328; the locus ESD was investigated in 78 healthy individuals). The analysis indicated that there were statistically significant deviations of the observed heterozygosities, g1, at 4 loci (GC, PI, C3, and ACPI) from the expected ones; h1 calculated from the data in the control group. The observed heterozygisities were higher than the expected ones at 3 loci (PI, C3, and ACPI), and at the GC locus. the observed heterozygosity being lower than the expected one. Comparing the observed heterozygosities. g1, within the loci, by using Fisher's exact test revealed significant differences between the groups of patients and healthy controls at the same loci, which showed significant differences between the observed and expected heterozygosities. There were no differences between the groups of patients by the observed heterozygosities. The mean expected heterozygosity were h = 0.386 +/- 0.056. The mean observed heterozygosity, were g = 0.415 +/- 0.037, 0.402 +/- 0.061, 0.371 +/- 0.055 in Groups 1 and 2 and in the controls, respectively. There were no differences between the mean expected and obsorved heterozygosities or between the mean observed heterozygosities in the three groups under study. It is proposed that a single locus rather than the mean heterozygosities should be used as a generalized nonspecific measure of genetic control over diseases while the former can show the involvement of a specific marker locus to develop a disease, the latter can simply veil the effects of each of the loci alone. Thus, the findings produce strong evidence for that there is a genetic control in the development of pulmonary tuberculosis. PMID- 11924121 TI - [Types and variants of cerebral circulation in healthy young people in clino stasis and anti-orthostasis]. AB - This paper presents the results of investigations of 199 apparently healthy individuals aged 18-22 years. Analysis of the findings yielded concepts of the types of cerebral circulation--the types of cerebral microcirculation and the variants brain volumia--the variants of summarized pulse blood filling of this organ. To define the types and variants of cerebral hemodynamics makes it possible to predict the magnitude and direction of brain vasomotor responses aimed at optimizing arterial blood stream into the region and venous blood outflow under antiorthostasis. The types and variants of cerebral circulation that most and least favourable as to a risk for regional venous congestion. The views of the types and variants of cerebral circulation open up fresh opportunities for in-depth study of the most important parameters of cardiovascular performance. A typological analysis of cerebral circulation can find a wide application in clinical practice for "the passportization" of the most important physiological parameters of regional blood flow. PMID- 11924120 TI - [Integrative study of bronchial asthma in perimenopausal women]. AB - The paper presents results of complex study of the specific features of the course of bronchial asthma (BA) in the peri- and early postmenopausal periods, including investigations of hormonal and immunological statuses, bone tissue mineral density and blood lipids, and evaluates the effects of peri- and early postmenopause and climacteric syndrome of varying severities on the course of BA. A drug test using estrogens has been developed to evaluate their effects on bronchial airflow. It has been shown that estrogens do not negatively affect bronchial airflow in most patients with BA in the peri- and early postmenopausal periods. The efficiency of replacement hormone therapy with estrogen-progestogen drugs has been for the first time estimated in these patients. Treatment with the estrogen-progestogen drug clinonorm increases the effects of endogenous cortisol, which makes it possible to reduce a dosage of glucocorticosteroids and to increase the mineral density of bone tissue. PMID- 11924122 TI - [Lectin-dependent platelet aggregation and agglutination in patients with unstable angina]. AB - It is necessary to study the membrane receptors of platelets to understand the mechanism of their aggregation for effective treatment of patients with ischemic heart disease (IHD). The aim of this paper is to estimate the significance of carbohydrate residues of the glycoprotein receptors of platelet membranes and thrombogenesis in patients with unstable angina (UA). A total of 142 patients with UA and 20 healthy individuals were examined. The carbohydrate structure of platelet membranes was studied using lectins. Lectin-dependent platelet aggregation in patients with US was found to be associated with the activation of intracellular systems and to be fibrinogen/GP IIb/IIIa-dependent. The patients with US had elevated levels of fusoco-, N-acetyl-D-galactosamine-, and mannosamine-specific carbohydrate residues onto the platelet membrane surface. Higher platelet capacity for agglutination and aggregation by altering the carbohydrate specificity of GP IIb/IIIa receptors may be one of the causes of thrombogenesis in patients with US. PMID- 11924123 TI - [Use of the multi-component VP-4 vaccine to prevent acute respiratory diseases at preschool children's establishments]. AB - The multicomponent VP-4 vaccine from Klebsiella pneumonia, Staphyloccocus aureus, Proteus vulgaris, Escherichia coli antigens was used to prevent acute respiratory diseases (ARD) at preschoolchildren's establishments. Prevention was effected during an influenza epidemic with a drastic rise in the incidence of ARD. The VP 4 use involved 3 nasal and 6-9 oral administrations of the drug at an interval of 3-4 days. The duration of the course was 26 +/- 4 days. The preventive action of VP-4 on the incidence of ARD was followed up for 14 months after vaccination. One group comprised 89 vaccinated children, a matched group included 69 children whose age was 2 to 6 years. Immunoprophylaxis was found to promote a reduction in the number of children who had 4 ARD episodes or more during the whole follow-up. Administration of VP-4 prevented the development of repeated episodes ARD in a group of children aged 2-3 years and in frequently ill children who had 3 infections or more in the previous year. The slight reactogenicity of the drug and its long-term preventive effect allow this method of immunoprophylaxis to be recommended for frequently ill children, for those at high risk, and for those in the junior age group (under 4 years) before going to preschoolchildren's establishments. PMID- 11924124 TI - [Affinity sorbents having magnetic properties in the clinical picture and diagnosis of communicable and non-communicable diseases]. AB - Affine magnetic sorbents which have no analogs in the practice of our country have been for the first time developed for the rapid diagnosis of various life threatening diseases (plague, cholera, anthrax, glanders, meliodosis, tularemia, leptospirosis, dysentery, viral hepatitis A) and for the identification of their causative agents. The efficacy of new magnet-controlling test systems has been repeatedly confirmed by their applications in epidemiological events and emergencies: in the epidemiological surveillance of viral hepatitis A in Stavropol and in the Caucasian Mineralnye Vody towns, Stavropol Territory (1994), in the identification of cholera patients, in the detection of transmission factors, when monitoring during large epidemic out-bursts of cholera in Stavropol (1990), Daghestan (1994), as well as in the microbiological monitoring during military conflicts in the Chechen Republic (1995). The application of the sorbents has shown that their sensitivity is 4-5 times as much as that of conventional serological assays. In addition, biotechnologies for the production of polyacrylamide and composite aluminosilicate affine immunosorbents with magnetic properties have been developed. They have been used as the basis for designing immobilized granulated antigen reagents for the immunodiagnosis, differential diagnosis, evaluation of the time course and severity of a disease, the efficiency of therapy in patients with systemic scleroderma, proliferative arthritis, systemic lupus erythematosus, juvenile rheumatoid arthritis, osteochondrosis. PMID- 11924125 TI - [Effects of medium-weight molecules isolated from plasma of healthy and burnt animals on the cellular composition of cultured bone marrow erythroblastic islets]. AB - The so-called median-weight molecular fractions (M, B = 0.5-1.5 kDa) exerting a marked hemopoietic effect in the cultured bone marrow erythroblastic islets were isolated from the plasma deproteinates of healthy dogs and animals with acute inflammation (thermal skin burn) by Sephadex G-15 gel chromatography. Acute inflammation is attended by the occurrence of blood medium-weight molecular components that suppress erythropiesis and by the concurrent high rate of granulocytopoiesis in the cultured bone marrow erythroblastic islets. Medium weight molecular fractions from the blood of healthy dogs substantially were found to suppress erythropoiesis and to stimulate granulocytopoiesis in the cultured erythroblastic islets. PMID- 11924126 TI - [Molecular genetic methods for the detection of rifampicin-resistant Mycobacterium tuberculosis strains]. AB - RCR-heteroduplex (GDA) and chip methods were used to detect rifampricin-resistant (RR) and rifampicin-sensitive (RS) Mycobacterium tuberculosis (MTB) in the samples from patients (sputum) and in the clinical isolates of MTB from these patients (MB/BacT liquid medium and Lowenstein Jensen's (LJ) solid medium. The efficiency of detecting RR and RS of MTB (from the sputum) is 100 and 92.3% in the chip and GDA tests, respectively. Correlations between GDA (sputum) and drug test (LJ) were 91.7%, that of chip (sputum) and drug test LJ, 88.5%, chip (sputum) and chip clinical isolates (LJ), 100%. The efficacy of GDA and chip in the detection of RR of MTB strains is under discussion. PMID- 11924127 TI - [Orotic acid as a metabolic agent]. AB - The paper reviews clinical and experimental studies into the mechanisms of action of orotic acid (OA). OA has been shown to take an active participation in metabolic processes in the body. As a pyrimidine precursor, it plays a key role in the biosynthesis of nucleic acids and protein, regulates water-salt exchange, by increasing diuresis and reducing the volume of extracellular fluid. OA is also a cellular fixative of magnesium by producing pronounced antiarrhythmic, vasodulator, and cardioprotective effects. OA has ascertained to stimulate erythro- and leukopoiesis. The involvement of OA in metabolic processes explains its cardio- and neuroprotective effects. By enhancing the resistance of myocytes to ischemia, OA favourably affects the clinical course of myocardial infarction and on manifestations of heart failure. OA has been noted to have an angioprotective action and to play an important role in the energy provision of the hypertrophic myocardium, by increasing its contractility. The ability to enhance the functional reserves of the heart adapted to higher exercises accounts for its use in sportive medicine. When there are emergency emotional and vestibular stimuli, OA drugs show an antistressor actions and are effective in treating patients with borderline nervous and mental disorders. Whether OA can be used to treat gastrointestinal diseases is to be clarified. PMID- 11924128 TI - [The spectrum and level of low-weight molecular compounds of microbial origin in familial Mediterranean fever]. AB - Familial Mediterranean fever (FMF) is a disease whose etiopathogenesis is not clarified yet. The infectious theory of FMF has not been confirmed either. Nevertheless, the involvement of microbes in the trigger of an inflammatory process cannot be excluded since today's well-known pathogenetic processes in FMF are to be directly related to the key cells of an inflammatory response. According to the existing concept, homeostasis of small molecules originating from microbes (SMOM) in healthy individuals is achieved due to adequate immune system function with the preserved biocenosis of a macroorganism whose disturbance with resultant immune shifts triggers a cascade of inflammatory reactions in the body. An attempt was taken to reveal the participation of microorganisms at the onset of an inflammation in case of FMF by using chromatographic mass spectrometry to detect chemical components of microorganisms and their vital activity products. The method allows one to screen a large number of microbial markers in a clinical sample. Pronounced impairments in the homeostasis of non-protein SMOM were found in the blood of examined patients with FMF (n = 16). There was a uniformity of deviations from the normal values in all the examinees in the episodes and episode-free periods. These qualitative and quantitative deviations basically differ from the direction of changes with other diseases (n 59) or healthy individuals (n = 18). All significant deviations affect non-traditional participants of an inflammatory process in the host. The similar microecological breakages in the human body and their consequences have not been earlier detected and investigated. The findings show it necessary to continue studies to receive an answer to the question as to whether the detected homeostatic features of SMOM in patients with FMF are primary or what is their role in the etiopathogenesis. PMID- 11924129 TI - [Prospects of experimental and clinical determination of 8-hydroxy-2 deoxyguanosine as a biological marker of oxidative stress]. PMID- 11924130 TI - [Current views on the role and composition of intestinal microflora in healthy adults]. PMID- 11924131 TI - [Epidemiological surveillance of nosocomial pyo-septic infections]. AB - The paper analyzes the parameters of epidemiological surveillance of nosocomial pyoseptic infections (NPSI) at surgical hospitals. It considers its basic concepts and presents the authors own findings. Particular emphasis is laid on guidelines for detecting NPSI cases and on epidemiological surveillance. A method of a follow-up is proposed to solve problems in the informational and analytical support of surveillance. PMID- 11924132 TI - National Occupational Standards for healthcare scientists. PMID- 11924133 TI - The suitability of ISO 9001 as a quality system for a medical illustration department. AB - The Medical illustration Department within Bradford Hospitals NHS Trust is typical of many such units in the UK that are facing increasing competition from the commercial sector and Private Finance Initiatives, and at the same time, being driven by NHS initiatives towards having a demonstrable quality system. It was considered that having a recognized quality management system would ensure that the service provided was more efficient and effective, improve quality, guard against competition, be an asset if the organization was benchmarked against others, and provide evidence of quality assurance to our customers. This paper summarizes the history of quality management, discusses a number of quality systems in the NHS in general, and in Bradford Hospitals NHS Trust in particular, and puts forward the reasons for choosing ISO9001. Finally, the process leading to accreditation is described. PMID- 11924134 TI - Qualifications--credit where credit is due. PMID- 11924135 TI - Making the change. PMID- 11924136 TI - Chemonucleolysis of lumbar disc herniation with ethanol. AB - PURPOSE: To demonstrate the efficiency of the treatment of the lumbar disk herniation (LDH) with absolute alcohol. MATERIALS/METHODS: From June 1997 to September 2000, 118 patients with 126 LDH, 69 males and 49 females, aged 19 to 77 (mean age: 51.1 years) were treated with absolute alcohol nucleolysis. In 75 cases, the LDH were parasagittal, in 30 foraminal, in 17 medial and in 4 extraforaminal. In most cases, the sensitive symptoms and motor or reflex abnormalities corresponded to the lumbar disc herniation level. Nucleolysis was made in ambulatory condition and each patient underwent general anesthesia. All procedures were performed in surgical conditions and controlled under digital fluoroscopy. The disc puncture was laterally made at the junction of middle and posterior thirds of the disc, under strict lateral fluoroscopic control. A discogram was obtained before ethanol injection. Injection dose of absolute alcohol was 0.4 ml. The realization time was about 15 minutes. RESULT: Total improvement of symptoms was obtained in 97.55% of cases. Two patients remained with low-back pain and the failure treatment rate was 0.84% (1 case). CONCLUSION: Nucleolysis with ethanol is a very effective, safe and low cost treatment for any kind of LDH, that allows to treat several levels during the same procedure, and can be repeated several times. Nucleolysis with ethanol can be made in ambulatory condition and be proposed to patients who refuse surgical treatment and have allergic background. PMID- 11924137 TI - Dysembryoplastic neuroepithelial tumors: CT, MR findings and imaging follow-up: a study of 53 cases. AB - PURPOSE: To evaluate CT and MRI features and long term imaging follow-up of a large series of dysembryoplastic neuroepithelial tumors (DNTS). PATIENTS AND METHODS: We retrospectively analyzed CT (100%) and MR imaging (83%) findings of 53 patients with complex (n = 14), simple (n = 6) or non specific histological forms (n = 33) of DNTS. All patients underwent epilepsy surgery for the treatment of drug resistant partial seizures. Preoperative radiological follow-up from two to 10 years (81%) and a post-operative follow-up from one to 13 years (92%) were available. RESULTS: DNTs are intracortical tumors with no mass effect and no peritumoral edema. An associated deformity of the overlying skull was observed in 44% of the 34 patients with a cortical lesion of the convexity. We found a contrast enhancement of the lesion in 21% of cases, a calcic hyperdensity in 36% of cases and a cystic part in 7.5% of cases. DNTs were hypodense (82%) on CT examinations and had a decreased signal on the T1 Weighted Images (95%) and a hypersignal in T2 Weighted Images (100%) on MR imaging. Eighty-one percent of patients had a mean preoperative radiological follow-up of four years and the tumor was stable in size in all cases; 92% of patients had a mean post-operative radiological follow-up of 4.5 years and no recurrence was seen. CONCLUSION: Three radiological features of DNTs are helpful for the diagnosis: cortical location, absence of mass effect and no surrounding edema. Clinical, radiological and histopathological findings have to be considered together in order to assess the diagnosis and to differentiate DNTs, which are stable lesions from gliomas. PMID- 11924138 TI - [What is your diagnosis? Sturge-Weber syndrome]. PMID- 11924139 TI - [Imaging of a historical case of vertebral hydatidosis]. AB - Radiological, computed tomography and magnetic resonance imaging findings in a case of spinal hydatidosis are reported. PMID- 11924140 TI - Pilocytic astrocytoma: unusual feature. AB - Juvenile pilocytic astrocytoma is a well-defined brain tumor. It most often occurs in children and young adults. It is located in the posterior fossa and has typical imaging features, associating cystic and strongly contrast enhancing mural nodule. After complete surgical removal, its prognosis is excellent. Adult cases are seldom observed. They develop almost exclusively within the cerebral hemispheres and share the same imaging and prognostic characteristics as the pediatric forms. We describe the case of a 42-years-old man presenting with a huge heterogenous posterior fossa lesion. Histopathological analysis of the lesion after surgical resection diagnosed a juvenile pilocytic astrocytoma. These peculiar location and imaging features in an adult patient may be misdiagnosed for infectious lesions and must be recognized. PMID- 11924141 TI - Spinal epidural angiolipoma. AB - Spinal epidural angiolipoma is a rare benign tumor predominantly located in the mid-thoracic region. The authors report a case of spinal epidural angiolipoma in a 36-year old woman who presented with subacute paraplegia. Clinico-pathological and MRI findings of this uncommon tumor are discussed. PMID- 11924142 TI - [Diagnosis and follow-up of pseudo-meningiomatous metastasis of prostatic cancer: value of MRI]. AB - We present the case of a patient with neurological signs, who underwent a CT scan then a MRI which displayed a meningiomatous lesion. The general and biological examination revealed a metastatic prostatic carcinoma, and the meningeal lesion was considered a secondary lesion. Follow-up during hormonal therapy showed the regression of the meningeal lesion and thus confirmed our hypothesis, which avoided surgical biopsy. PMID- 11924143 TI - [Calcified cerebral hydatid cyst. Report of a case]. AB - The cerebral localization of the hydatid cyst is still rare (1 to 4% of the cases). The calcified cerebral hydatid cyst is exceptional. We report one case collected in our department. It is about a patient of 28 years old, that the case histories consist in convulsive attacks since the age of 8 years old, hospitalized because of a left hemiparesis with progressive installation. The cerebral computerized tomography showed a right parieto-occipital that is largely calcified. During the operation, a calcified hydatid cyst discovered with various daughter vesicles and a cerebral gliosis, the latter has been extracted entirely. The evolution has been favourable with improvement of the left hemiparesis. The calcified hydatid cyst of brain is still exceptional, its symptomatology is the same as the safe CHC, but it poses some diagnostic and therapeutic problems. PMID- 11924144 TI - [Atypical cerebral lymphoma with ocular onset: report of a case]. AB - A 72 year-old woman is hospitalized and explored in the neurological department because of dementia discovered after a vitrectomy performed to treat a relapsing chronic uveitis. MRI examination shows atypical hyperintense white matter lesions on T2 weighted images related to Lyme disease. Worsening of clinical status, despite appropriate medical treatment and apparition of enhanced nodules MR images rules out the diagnosis of lyme disease and is attributed to brain metastases. The search for primitive tumor is not contributive and a brain biopsy is performed. It discloses a B cells non-hodgkin lymphoma. This case report stresses two points: first, a lymphoma must be one of the diagnosis to evocate if imaging shows an enhancing nodule, or extensive hypersignal of the white matter, second: it emphasizes the value of vitreous analysis searching for lymphoma during chronic uveitis associated to neurological symptoms. PMID- 11924145 TI - Leucoencephalopathy after heroin inhalation. A case with partial regression of MRI lesions. AB - We report the case of a 41 year old patient who developed a severe cerebellar ataxia. MRI findings were suggestive of myelin damage with symmetrical involvement of the cerebellar hemispheres and, to a lesser extent, the decussation of the superior cerebellar peduncles, the corticospinal tracts and the centrum semiovale. He had been inhaling heroin for the last 5 years. Two years after stopping heroin, he showed clinical improvement with partial regression of the MRI lesions. MRI findings of leucoencephalopathy after heroin inhalation are well described in the literature, however longitudinal studies are rare. It is the purpose of this report to show that clinical and MRI features can be characteristic of this leucoencephalopathy and that regression of white matter lesions can be seen after heroin withdrawal. PMID- 11924146 TI - [Contribution of imaging to the diagnosis of Darier-Ferrand's cranio-facial dermatofibrosarcoma. Report of 2 cases]. AB - Dermatofibrosarcoma protubans of Darier-Ferrand is observed in about 0.1% of al malignant skin tumors. Head and face localization in uncommon, estimated to account for about 1 to 10% of all lesions. Relapse is frequent and metastasis in uncommon. Diagnosis is often made late. Surgery is the basic treatment. We report two cases of dermatofibrosarcoma of Darier-Ferrand with head and face localizations. In one case, the tumor extended inside the skull and spread to the superior longitudinal sinus. This is the only such reported case found in the literature. CT scan is more useful for studying the bony tables and the cranial content. MRI provides precision for soft tissue tumors and angio-MRI is particularly useful for analysing with accuracy vascular relations of the tumor. PMID- 11924147 TI - [Nosocomial pneumopathy acquired under mechanical ventilation. Suspicion criteria, bacteriologic diagnosis, and remission criteria]. AB - In intensive care patients, pneumonia acquired under ventilatory assistance is the second most frequent nosocomial infection after urinary tract infection. Duration of ventilatory assistance and stay in the intensive care unit are both increased in this leading cause of death due to nosocomial infection. A large number of studies and national and international consensus conferences have been devoted to determining precise criteria leading to clinical suspicion of pneumonia acquired under ventilatory assistance and the appropriate elements for bacteriological diagnosis. The criteria retained in these different studies are neither precise nor reproductable. Based on data in the literature and our their clinical experience the members of the Outcomerea association constituted working groups to elaborate a set of guidelines that could be applied in routine clinic in response to three questions: 1) what criteria are suggestive of pneumonia acquired under ventilatory assistance, 2) what bacteriological tools are needed to confirm diagnosis, 3) how and why should cure be defined. We present a review of the literature and the conclusions of the working groups. PMID- 11924148 TI - [Positron emission tomography (PET) with [18F]-FDG in bronchopulmonary cancer and its impact on medical decision at the time of diagnosis, staging, or recurrence evaluation]. AB - Clinical usefulness of [18F]-FDG imaging, performed by means of a dedicated or a "hybrid" PET machine, has been recognised in France since November 1998. Among the clinical indications, three major clinical settings of lung cancer have been included: characterisation, staging and detection of recurrences. After a brief presentation of the PET scintigraphic imaging modality, authors report on the experience of the nuclear medicine team of Hospital Tenon and summarise the results in literature. For tumour characterisation, a recent meta-analysis obtained a 96% sensitivity, a 73% specificity, a 91% positive predictive value and a 90% negative predictive value, the performances being better for lesions greater than 1 cm. For staging, an increase greater than 15% both in sensitivity and specificity has been observed with dedicated or "hybrid" PET versus CT for N staging. Detection of distant metastases was also more accurate using [18F]-FDG. A similar increase was observed in the detection of recurrence, in accordance with our study; some authors described even better results. A better anatomical delineation of the lesions detected with FDG can be achieved by means of image fusion with CT; this technique is likely to develop as a routine tool in the near future. Finally, FDG imaging led to modification of patient's management in 37% of the cases according to a recent meta-analysis versus 53% of the cases in our retrospective survey concerning the first year of installation of a dedicated PET machine. This rate was equal with dedicated PET and with CDET. In 46% of the cases an inter-modality change occurred, and in 7% an intra-modality change consisting mainly in adaptation of the surgical procedure. As soon as the FDG examination became available, its clinical impact, in the French medical context, appeared to reach the highest values that were published internationally. PMID- 11924149 TI - [Pulmonary circulatory amputation in chronic respiratory insufficiency. Simple, anodyne, and fast evaluation]. AB - In chronic respiratory insufficiency, vascular bed amputation is a key element of the prognosis. The assessment of the carbon monoxide transfer capacity of the lungs is often impossible for technical reasons. However, the measurement of CO uptake (VCO), normalized by the reject of CO2 (VCO2) is a straightforward and repeatable measure for most healthy individuals. Its deficit is most likely due to either a circulatory impairment of the "gas-exchanging organ" and/or an alveolary ventilation failure. AIM: To define the relationship between hypoxemia and VCO/VCO2 deficit. METHOD: The subjects were classified into four causes of the disease: pneumonectomy, kyphoscoliosis, emphysema and chronical bronchitis. VCO/VCO2 was measured with an open circuit and simultaneous arterial blood-gas analysis. RESULTS: With a few exceptions, VCO/VCO2 deficit was always observed and more important that hypoxemia. VCO/VCO2 and PaO2 values were not related in the case of emphysema, but strongly correlated and proportionally impaired in both pneumonectomy and kyphoscoliosis cases. CONCLUSION: This rapid, simple and well-tolerated measurement of VCO/VCO2 could provide a much-improved method of prognosis over the current CO measures. PMID- 11924150 TI - [Role of pharmacists in the management of asthma in Africa. Survey among pharmacists from the city of Abidjan]. AB - This questionnaire survey was conducted to assess delivery of asthma drugs in community pharmacies in Abidjan, Ivory Coast. Participation rate was 88.69%. All participants stated they provide care for asthma patients either during acute episodes or between acute episodes. It was found that a considerable number of patients come to community pharmacies for care without prior medical prescription. In case of acute episodes, care is provided by the pharmacist in 59.80% of the cases. Aerosol devices are the most widely used although the technical procedures required for correct inhalation were found to be poorly understood by pharmacists. Slow-release corticosteroids are widely delivered for chronic care. Pharmacists also provide information and advice on drug compliance and concerning the importance of regular medical care and eviction of allergens. PMID- 11924151 TI - [Radiographic and bacteriologic data on pulmonary tuberculosis in Bamako as a function of HIV status]. AB - This prospective study included patients treated for tuberculosis between January 998 and April 1999 in the pneumology unit of the Point G Hospital in Bamako. The purpose was to analyze chest x-ray and bacteriological findings in HIV+ and HIV- patients. All patients had clinical and radiographic signs suggestive of tuberculosis with one or several sputum samples and HIV serology. Among the 127 patients, 36 were HIV positive (28.3%). There was no sex predominance in the HIV+ patients (p = 0.3). The most common radiographic aspect associated nodules and infiltration irrespective of the immune status (p = 0.014). Characteristic features of the chest x-rays in HIV-positive patients were: frequent military forms and rare cavernous forms (p = 0.007). Sputum tests were positive more often in HIV-negative than HIV-positive patients (p = 0.0003). The diagnosis of tuberculosis was retained in 11.7% of the patients with negative microbiology despite repeated samples. A normal chest x-ray was observed solely in one HIV negative patient. PMID- 11924152 TI - [Chordoma: atypical mediastinal mass. Report of a case]. AB - Chordoma is a rare tumor which develops from remnant notochord tissue. Sacro coccygeal and spheno-occipital localizations predominate. We report a thoracic chordoma of the fifth thoracic vertebra with mediastinal expansion in a 70-year old woman. Pathology confirmed the diagnosis revealing a lobulated architecture, presence of physaliphorous cells and intracellular mucoid substance. Immunohistochemistry can also be very helpful in atypical cases: positive for anti-cytokeratine, antivimetin and anti-protein S100 antibodies. Spontaneous outcome is fatal. Early and complete resection is the only way to improve prognosis. PMID- 11924153 TI - [Pseudo-tumoral aspergilloma]. AB - Aspergilloma is the most common form of pulmonary aspergillosis, generally developing pre-existing lung cavities. Fiberoptic bronchoscopy is required in case of hemoptysis. We report the case of a 74-year-old man with pulmonary aspergilloma where fiberoptic bronchoscopy visualized mycetoma and cavitation. Visualization and biopsy of the fungus ball during fiberoptic bronchoscopy is rare. PMID- 11924154 TI - [Bilateral excavated nodular pneumopathy: amebiasis? Report of a case]. AB - Pleuropulmonary amibiasis is generally secondary to hepatic amibiasis with migration into the thorax. Direct pulmonary involvement is exceptional. We report a case of pulmonary ambiasis without associated liver involvement causing multiple bilateral pulmonary lesions. PMID- 11924155 TI - [Serous fibrinous pleurisy disclosing hydatid cyst of the liver fistulizing into the pleura]. AB - Hydatic cyst of the liver remains a serious public health problem in Morocco and several other Mediterranean countries. This benign affection can cause serious sometimes fatal complications such as cyst rupture into the biliary ducts or the thorax. We present the case of a patient with serofibrous pleurisy and recall the often late diagnosis of hydatic cyst. Biliary pleural fistulization is a rare but serious complication. PMID- 11924156 TI - Protecting the privacy of patients' health information. Summary of the final regulation. PMID- 11924157 TI - Release of behavioral health, developmental disabilities, HIV, and substance abuse information: guidelines for legal compliance. PMID- 11924158 TI - Kigris. FDA approves first biologic for sepsis. PMID- 11924159 TI - Coreg. New indication for heart failure drug. PMID- 11924160 TI - Tracleer. Tablets ease symptoms of rare lung disorder. PMID- 11924161 TI - Protonix. First i.v. proton pump inhibitor approved. PMID- 11924162 TI - Spectracef. New cephalosporin antibiotic approved. PMID- 11924163 TI - Treating HIV. On-and-off therapy works--and cuts costs too. PMID- 11924164 TI - Preparing for bioterrorism. PMID- 11924165 TI - How to change a PICC dressing. PMID- 11924166 TI - Putting a patient with diabetes in the driver's seat. PMID- 11924167 TI - What you should know about chronic grief. PMID- 11924168 TI - Facing neuro assessment fearlessly. PMID- 11924169 TI - Expanding the perioperative role. The surgeon's assistant. PMID- 11924170 TI - Consenting to perioperative visitors. PMID- 11924171 TI - Profile of the events team. PMID- 11924172 TI - Preparing for Europe. PMID- 11924173 TI - The it boy. PMID- 11924174 TI - Law and professional practice. Accountability and implications. AB - The role of the operating theatre nurse has expanded greatly in recent years and mistakes or errors are inevitably made. The majority of mistakes are usually easily corrected or go unnoticed, but those that aren't can produce substantial implications for the patient and the practitioner. PMID- 11924175 TI - [COPD: 3 challenges, one global answer]. PMID- 11924176 TI - [Occupational chronic obstructive pulmonary disease (COPD)]. PMID- 11924177 TI - [A plea for standardized thoracoscopy in malignant pleural mesothelioma]. PMID- 11924178 TI - [Positron emission imaging and lung cancer]. PMID- 11924179 TI - [Invasive pulmonary aspergillosis in hematology]. PMID- 11924180 TI - [Value of 18FDG-CDET in the evaluation of operable bronchial cancer]. AB - The purpose of this study was to investigate the utility of 18fluorodeoxyglucose (FDG) coincidence detection position emission tomography (CDET) in the evaluation of metastatic mediastinal lymph nodes in patients with potentially operable non small-cell lung cancer (NSCLC). A prospective study was performed in thirty patients with newly suspected NSCLC. Thoracic computed tomography (CT), FDG CDET, and invasive surgical staging were performed in patients. Blinded prospective interpretation was performed for each test and compared to pathological staging obtained by mediastinoscopy and/or by thoracotomy. Patients were followed for six months to detect occult metastases. The sensitivity and specificity of CDET for the detection of mediaStinal lymph nodes were 75% and 94.4% respectively. The corresponding value for CT were 50% and 80.9%. Three patients with N1 disease were classified as N0 by CDET. With regard to definitive surgical node staging, CDET could identify nodal disease in 26 patients and CT only in 18 patients (n = 30). FDG full-ring positron emission tomography (PET) is the most accurate non invasive method for the detection and staging of lung cancer. In addition, FDG CDET shows high accuracy for the detectability of pulmonary lesions with a diameter at least 2 cm and the evaluation of lymph node in NSCLC. PMID- 11924181 TI - [Invasive aspergillosis in the leukemic patient]. AB - Invasive pulmonary aspergillosis (IPA) remains a life threatening complication in immuno-compromised and especially in neutropenic patients. We report our experience in the diagnosis and therapeutic management of IPA in 8 patients with acute leukemia. All patients were neutropenic (PNN < 100/mm3, mean duration = 37 days) when IPA was diagnosed. Clinical signs included fever above 39 degrees and cough in all cases, chest pain in 4 cases, hemoptysis in 3 cases, rales in 5 cases. Chest x ray showed one lesion in 4 cases and multiple lesions in 4 cases. The diagnosis of IPA was established by bronchoalveolar lavage (BAL) in 5 cases, tissue biopsy in one case, positive sputum in one case and it was highly probable in one case. Thoracic computed tomographic (CT) scans were preformed after diagnosis confirmation of IPA and showed one or multiple lesions with air crescent signs. Serological tests were positive in 4 cases late in the course of IPA. All patients were treated with i.v. Amphotericin B. Outcome was favorable in 5 cases and three patients died by massive hemoptysis (in two cases) and systemic aspergillosis (in one case). Early diagnosis and appropriate treatment are essential to improve IPA prognosis. PMID- 11924182 TI - [Respiratory risk in carpenters and cabinet makers]. AB - They are many risks relating to the wood; they are caused by natural components of wood, products of conservation, chemical agents and parasites of wood. We have carried out a retrospective survey which concerned exposed workers and controls in twenty small handicraft workshops in the joiners' souk of Marrakesh, it has enabled us to evaluate the prevalence of the clinical symptoms and disorders of respiratory function in 242 exposed subjects to the wood dust and 121 controls. This enquiry consisted of a questionnaire (European Coal and Steel Community: ECSC and the World Health Organisation: WHO), a clinical examination and a spirometry. Sixty-one point nine % of those exposed had clinical respiratory symptoms versus only 21.5% of controls. Rhinitis, asthma, conjunctivitis, chronic bronchitis and dermATitis were significantly more frequent in those exposed than among the non-exposed, with respectively 55.8%, 14.5%, 24.8%, 21.1% and 12.8% versus 16.5%, 6.6%, 8.3%, 5.8% and 4.9%. Exposure was the cause of respiratory symptoms because among non-smokers, exposed workers were more symptomatic than controls. Smoking exhibited a potentializing effect on airborne occupational contaminants because among exposed workers disorders were 1.8 times more frequent in smokers than non-smokers. A variable degree of respiratory obstruction was found among 30.1% of the exposed individuals versus 12.4% of the unexposed subjects. The effect of exposure was certain because among the non-smokers, 15% of exposed subjects had altered respiratory function versus 4% of unexposed persons. It is imperative to implement an occupational health service and to develop means for collective and individual prevention to maximally reduce the risk. PMID- 11924183 TI - [18FDG-PET applications in thoracic oncology]. AB - PositIon emission tomography using 18-fluorodeoxyglucose (FDG-PET) is an imaging technique based on metabolic criteria rather than morphological criteria. We discuss the contribution of this technique in the field of pulmonary oncology and we summarize our work which has demonstrated the performance capacity of PET: to discriminate the malignant nature of a solitary pulmonary nodule; to improve sensitivity over CT for mediastinal assessment in non-small cell lung cancer; to seRach for distant metastasis in patients with non-small cell lung cancer; to complement CT imaging to better dissociate tumor residue or recurrence from post therapeutic sequelae in non-small cell lung cancer. Finally, we present different expected progress for the future. PMID- 11924184 TI - [Mechanical ventilation-related variability of stroke volume. Clinical evaluation and therapeutic implications]. AB - Mechanical ventilation induces cyclic changes in left ventricular stroke volume. These variations are mainly related to the expiratory decrease in left ventricular preload following the inspiratory decrease in right ventricular filling and ejection. Therefore, the magnitude of the respiratory changes in left ventricular stroke volume reflect the sensitivity of the heart to the cyclic changes in preload induced by mechanical insufflation. At the bedside, the respiratory changes in left ventricular stroke volume can be assessed by the analysis of the arterial pressure (arterial catheter) or aortic blood velocity (echocardiography) wave forms. The respiratory changes in arterial pressure and in aortic blood velocity have been shown to be accurate predictors of fluid responsiveness and of the hemodynamic effects of positive end-expiratory pressure. PMID- 11924185 TI - [Tracheal replacement: new perspectives]. AB - Tracheal replacement remains a real challenge in thoracic surgery. For tracheal resections greater than 5 to 6 cm, end to end anastomosis is not possible. Several solutions including prostheses, tracheal homografts and various autologous tissues have been proposed for tracheal replacement but all are associated with multiple complications and surgical difficulties. None of the numerous experimental studies have produced consistent results allowing for a standardized clinical approach. We propose the use of an autologous aortic graft for tracheal replacement. In a series of experiments, we have observed a progressive transformation of the arterial graft into a tracheal tissue after implantation. This observation raises interesting questions concerning histological adaptation and may offer new perspectives in human tracheal replacement. PMID- 11924186 TI - [Chronic respiratory failure in children. Evaluation and management]. AB - Chronic respiratory insufficiency in childhood can be caused by a heterogeneous group of diseases of which the management and prognosis differ. Bronchopulmonary dysplasia in infants and cystic fibrosis and neuromuscular diseases in older children, represent the most common causes of chronic respiratory insufficiency. The improvements in medical treatments, with the development of long term oxygen therapy and non invasive mechanical ventilatory assistance, have contributed to the increased survival and the better quality of life of these children. An objective evaluation of treatments, such as long term oxygen therapy and non invasive mechanical ventilation, are warranted to improve the indications and benefits of these therapies. PMID- 11924187 TI - [Hypoxemia secondary to inferior vena cava return into left atrium]. AB - The case of a right-to-left shunt-induced hypoxemia with an abnormal return of the inferior vena cava (AIVCR) into the left atrium (LA) is reported in a 30-year old male with cyanosis and polycythemia. The chest X ray and the lung CT scan was normal. Spirometry was normal but the transfert-CO coefficient (KCO) was lowered. Hypoxemia was observed at rest and worsening during exercise. The alveolo arterial oxygen tension difference under hyperoxia was increased (56 kPa). Contrast echocardiography (CEch) suggested the presence of an AIVCR with a right to-left shunt only observed by the inferior route. The inferior vena cava (IVC) angiography and the magnetic resonance imaging demonstrated an AIVCR characterized by a direct drainage of IVC in the left atrium. The good tolerance can be explained by the association of AIVCR with an inter-auricular septal defect resulting in a left-to-right shunt which partially corrected the right-to left shunt. After surgical treatment, arterial blood gases normalized, KCO remained low and CEch became negative. PMID- 11924188 TI - [Acquired right-to-left shunt with normal right heart pressure]. AB - Intracardiac right-to-left shunting through a patent ovale foramen (POF) despite normal right-sided pressures is an uncommon cause of dyspnea with posture worsening in adults. We report herein 6 cases which sum up etiological circumstances, diagnostic workup and management of this syndrome. The main reported etiopathologic mechanism is consistent with a change in the internal configuration of the right atria leading to preferential blood flow from inferior vena cava through patent foramen ovale. The treatment lies on surgical closure of the septal defect, which can now be performed using a noninvasive percutaneous procedure. PMID- 11924189 TI - [Fat embolism with lung hemorrhage]. AB - Post-traumatic fat embolism was disclosed by a picture of alveolar hemorrhage. Acute hypoxemia associated with dense bilateral pulmonary infiltrates was observed in a 21 year-old woman, 4 days after an accident with closed tibial fracture. Cruoric pulmonary thromboembolism was ruled out, as was an acute pulmonary edema. Neither infectious nor immunologic etiology was found. The diagnosis of alveolar hemorrhage was based on bronchoalveolar lavage. Lipid droplets in macrophages stained by "Oil Red O" established the relationship with fat embolism. The outcome was favorable. The association of fat embolism and alveolar hemorrhage has already been reported, but remains rare. PMID- 11924190 TI - [Meningeal metastasis of pleural mesothelioma]. AB - Meningeal metastases from malignant mesotheliomas are rarely observed. We report the case of a 54-year-old man with an asymptomatic pleural effusion and simultaneous sensitive and motor disorders of the right hemibody. A meningeal localization of a pleural malignant mesothelioma was discovered and confirmed by a comparative immuno-histological analysis. Here we present a differential diagnosis and a review of the literature to give prominence to diagnostic pitfalls in this rare disease. PMID- 11924191 TI - [Metastatic gastric calcifications and bronchial neuroendocrine tumor]. AB - We report the case of a 38 year-old man admitted for the treatment of a neuroendocrine carcinoma of the lung with multiple bone metastases. At the diagnosis, the serum biochemistry revealed an evidence of malignant hypercalcemia with acute renal impairment. At this point, a Tc 99-m MDP bone scan was performed and showed intense uptake throughout the gastric walls. The patient underwent a repeat bone scan after normalization of biochemistry; no more abnormal uptake was noted in the stomach. Hypercalcemia is an abnormality of the calcium metabolism frequently associated with malignant condition. Metastatic calcification results from increased accumulation of the calcium-phosphate salts in different tissues related to a local physiological alcalinity. Usually reversible, metastatic calcifications appear as various extraskeletal uptake at Tc 99-m MDP bone scan. PMID- 11924192 TI - [Pulmonary eosinophilia with excavated opacities possibly induced by salazosulfapyridine]. PMID- 11924193 TI - [Reactivation of tuberculosis after antiretroviral treatment in an HIV-infected patient]. PMID- 11924194 TI - [Current antibiotic therapy: lower respiratory tract infections in children]. PMID- 11924195 TI - [Current antibiotic therapy: exacerbated chronic bronchitis]. PMID- 11924196 TI - Nursing quality indicators. Research proves nursing's impact on patient care. PMID- 11924198 TI - Spotlight on psychiatric nursing: enlisting a unified force for the future of the profession. PMID- 11924197 TI - "I've got a right to sing the blues; or, recognizing depression in the elderly". PMID- 11924199 TI - In the hands of women. The female condom initiative. PMID- 11924200 TI - Where are the recruiters? PMID- 11924201 TI - Fetal heart assessments. Determining which technique to use. PMID- 11924202 TI - Silence is not golden. PMID- 11924203 TI - When the unexpected happens. AB - Last week I was working an evening shift when one of my clients had a serious unexpected incident. The client's family was present at the time and asked many questions and took notes. I felt I was being watched and judged. I was unsure about what information I should or could provide the family, but remembered someone once saying that it is better to say nothing in this type of situation. However, I felt it was important that I keep the family informed and offer empathy and care. Because I was unsure of what to do in this situation, I contacted an RNABC nursing practice consultant and reviewed our hospital policies and procedures. I know I did the right thing by keeping the family informed. PMID- 11924204 TI - CDC director to step down March 31. PMID- 11924205 TI - Federal task force considers condom distribution in prisons. PMID- 11924206 TI - Blood transfusion. Trial judge damaged credibility of expert witness. PMID- 11924207 TI - Illinois considers restoring AIDS drug assistance funding. PMID- 11924208 TI - Fast-food chain must pay damages for needlestick incident. PMID- 11924209 TI - Elizabeth Dole announces Senate run to replace Helms. PMID- 11924210 TI - CDC issues HIV prevention fact sheets for injection drug use. PMID- 11924212 TI - New HIV/AIDS center expands services to Native Americans. PMID- 11924211 TI - AIDS researcher Fauci off shortlist for NIH directorship. PMID- 11924213 TI - Bush appointees take helm at advisory council. PMID- 11924214 TI - Oregon physician under scrutiny for approving medical marijuana. PMID- 11924215 TI - CDC surveillance report notes slowdown in new HIV/AIDS rates. PMID- 11924216 TI - Nurses committed to strike goals: safe patient care, respect, equitable wages. PMID- 11924217 TI - Nursing at New York's Ground Zero. PMID- 11924218 TI - True leaders emerge at OHSU strike. PMID- 11924219 TI - OHSU nurses fight for future of nursing. PMID- 11924220 TI - Historic contract negotiated in Redmond. PMID- 11924221 TI - Breast cancer support resource available. PMID- 11924222 TI - Minimize exposure to DEHP. PMID- 11924223 TI - [Treatment of periodontitis associated with cyclosporin-induced severe gingival hyperplasia with regular mechanical plaque removal in a patient with renal transplantation (Case report)]. AB - A clinical case of a middle-aged kidney transplanted woman is presented. The woman has been taking Cyclosporin-A for over 13 years and has had advanced periodontitis and severe gingival swellings and gingival inflammation. The kidney transplant patient was treated and followed up for approx. four years. The treatment protocol included very thorough mechanical scaling and root planing, oral hygienic instructions and the regular professional maintenance program resulted in complete remission of the gingival overgrowth and stabilization of the periodontal condition. The gingival and periodontal conditions showed a continuing improvement over the time despite of the continuous CSA administration. PMID- 11924224 TI - [Effect of chewing sugar-free gum on dental caries]. AB - Previous in vivo studies have demonstrated that chewing sugar-free gum after eating reduces the development of dental caries. To investigate the extrapolation of these findings, a two-year clinical study was conducted on 547 schoolchildren in Budapest. Subjects in the "gum" (test) group were instructed to chew one stick of commercially available sorbitol-sweetened chewing gum for 15-20 minutes after meals, three times daily. The "control" group was not provided with chewing gum. After two years, excluding white spots, the "gum" group exhibited a 38.7% caries increment reduction compared to the "control" group. Including white spots, a corresponding 33.1% reduction was indicated. These results clearly suggest that even in a population with moderate caries prevalence and normal oral hygiene habits (including the use of fluoride dentifrices), an after-meal gum chewing regimen can significantly reduce the rate of caries development. PMID- 11924225 TI - [Extent of microleakage in relation to root canal morphology]. AB - The purpose of this study was to study with standardized methods the length and extent of the microleakage developing around the root canal fillings. The preparational errors were evaluated in twenty straight and curved root canal using computer graphic analysis. The step back technique was used for root canal preparation. The shape of the instrumented root canals were analyzed macroradiographically in order to evaluate unwanted procedural root canal deformities. The root canals were obturated with lateral condensation technique using AH-26 sealer and guttapercha points. After the teeth having been made transparent a silver nitrate penetration test was performed in vacuum and both the linear dye penetration and the extent of dye penetration surface area were measured from four directions with computer assisted digitized method. The linear dye penetration values were higher in straight root canals than in curved ones: 2.22 + 1.17 mm and 1.81 + 1.24 mm respectively. The dye penetration surface area was slightly also larger in straight than in curved root canals: 1.3 +/- 0.71 mm2 vs. 1.06 +/- 0.5 mm2. Nevertheless the differences were statistically non significant. In curved root canals the highest dye penetration values were measured at the inner curvature of the canals. This findings was also corroborated by a coincidence test. PMID- 11924226 TI - [Cleft lift and palate surgery from the viewpoint of orthodontics, maxillo-facial and reconstructive surgery--state of the art in Hungary]. AB - The author reviews the literature of cleft lip--and palate concerning on the incidence and treatment of these disorders. Describes the most often used surgical treatments, emphasises the importance of team work, details the surgical and orthodontic roles. The author reviews the treatment plan of cleft lip and palate applied at the Oral--and Maxillofacial Clinic of the Semmelweis University and suggests to introduce it as a general procedure. The author nominates to create a Hungarian Cleft Centre based on the declaration of EuroCleft project. PMID- 11924227 TI - [Diagnosis of Helicobacter pylori infection from dental plaque]. AB - The key role of Helicobacter pylori (Hp) in gastric inflammations is discussed extensively in the literature. In the present study in 140 school-children aged 12 years, Hp was traced in dental plaque by use of the polymerase chain reaction (PCR). Oral Hp was detected in the case of 11 children, where subsequent UBT did not confirm gastric Hp infection, however during family research in 8 of the 11 families at least one of the parents was found Hp positive. It was concluded that the non-invasive dental plaque PCR may play a significant role in the early indication of Hp infection. PMID- 11924228 TI - [Systemic antibiotic treatment in routine practice. Exacerbated chronic bronchitis, lower respiratory tract infections in children, acute otitis media, acute sinusitis in children, acute sinusitis in adults]. PMID- 11924229 TI - [Management of carcinomatous metastatic pleurisy with unknown primary neoplasm]. AB - The primary cancer remains unknown in about 7% of all cases of metastatic carcinomatous pleurisy. Thoracoscopic pleural biopsy provide good quality histology material. The pathology examination, including immunohistochemistry using a panel of well-defined antibodies can provide the diagnosis of neoplastic pleurisy of malignant carcinomatous proliferation, and help search for differentiation and rule out an identifiable primary carcinoma. When histology does not provide a satisfactory clue to the primary cancer, frequent origins include lung cancer, breast or ovary cancer in women, prostate cancer in men, and digestive tract cancer. The possible contribution of positron emission tomography remains to be evaluated. Several chemotherapy protocols can be proposed but the beneficial effect has been difficult to assess due to the heterogeneous nature of the histological forms treated. The carboplatin-paclitaxel-etoposide protocol appears to be promising in cases with an unknown primary cancer but there has been no specific assessment of this chemotherapy schedule in a subgroups of patients with carcinomatous pleurisy with an unknown primary cancer. Better understanding of the biological profile of highly metastatic cancers should be helpful in determining more targeted therapeutic schemes. PMID- 11924230 TI - [Endobronchial tuberculosis. Report of 21 cases]. AB - Since the advent of antituberculosis drugs, endobronchial tuberculosis has become an exceptional finding, often in a misleading context. Cases reported generally occur in patients with immune deficit. We report 21 cases of endobronchial tuberculosis in immunocompetent patients (mean age 37 years). Clinical and radiological signs were not specific. Bronchial fibroscopy was essential for certain diagnosis, particularly in budding forms. Early treatment reduced the risk of complications. Based on our experience with these 21 cases, we propose a discussion of the pathogenic mechanisms involved and the diagnostic difficulties encountered as well as the disease course and risk of complications that can be expected. PMID- 11924231 TI - [Pleomorphic adenoma with bilateral pulmonary metastasis]. AB - We report a new case of pleomorphous adenoma of the submaxillary glands with multiple lung metastases. Histological proof was obtained on the thoracoscopic surgical specimen. Clinically, this benign tumor presents as a malignant tumor. Diagnosis has been a subject of debate; surgical resection is indicated. Diagnosis is achieved by elimination in a patient with one or several nodules occurring in a context of recurrent pleomorphous adenoma. PMID- 11924232 TI - [Congenital cystic adenomatoid malformation of the lung. Prenatal diagnosis in a case]. AB - We report a case of congenital cystic adenomatoid lung malformation, type I in the Stocker classification, diagnosed at ultrasonography at 27 weeks gestation. The lesion stabilized as was the hydramnios. Vaginal delivery was uneventful. Early surgery with lobectomy led to a favorable outcome at 8 months follow-up. Congenital adenomatoid lung malformation is a rare finding. Prenatal ultrasound diagnosis has enabled early treatment and improved prognosis. PMID- 11924233 TI - [Intrathoracic textiloma caused by transdiaphragmatic migration after biliary surgery. Report of a case]. AB - Intrathoracic textiloma is very rare and the transdiaphragmatic migration after abdominal surgery is exceptional. The diagnosis was suspected by history of surgery and radiology (CT), the treatment is only surgical. We report a case of intrathoracic textiloma after biliary surgery 30 years before and we describe the clinical, radiological features and treatment of this severe complication including clinical and medico-legal consequences. PMID- 11924234 TI - [Classical palpebral edema]. PMID- 11924235 TI - [Methodology of clinical trials]. PMID- 11924236 TI - [Detection of lung cancer]. PMID- 11924237 TI - [Neoadjuvant treatment for early stages of non-small-cell lung carcinoma (NSCLC)]. PMID- 11924238 TI - [Adjuvant treatment for early stages of non-small-cell lung carcinoma (NSCLC)]. PMID- 11924239 TI - [Therapeutic management of non-small-cell lung carcinoma (NSCLC) of stage III A]. PMID- 11924240 TI - [Does treatment for malignant pleural mesothelioma exist?]. PMID- 11924241 TI - [Management of non-small-cell lung carcinoma (NSCLC) of stage III B]. PMID- 11924242 TI - [Phase III clinical trials concerning non-small-cell lung carcinoma (NSCLC) presented at the ASCO 2001]. PMID- 11924243 TI - [New therapeutic approaches in non-small-cell lung carcinoma (NSCLC), stages IV]. PMID- 11924244 TI - [New prospects in the treatment of non-small-cell lung carcinoma (NSCLC): new biological factors]. PMID- 11924246 TI - Risky business? Do infertility treatments damage babies' genes? Doctors used to think not. Now they are not so sure. PMID- 11924245 TI - [Lymph node excision in the surgical treatment of bronchopulmonary cancer]. PMID- 11924247 TI - Transplants. A womb of one's own. PMID- 11924248 TI - What's a guy to do? Most men who get prostate cancer don't die from it. So why get tested? Good question. PMID- 11924249 TI - Nursing heroes: caring at a time of national tragedy. PMID- 11924250 TI - Mandatory overtime: conflicts of conscience? PMID- 11924251 TI - Transitions in American hospitals: the necessary reshaping is taking place. "Turnaround" processes in organizations. AB - American hospitals are reshaping and redefining the way they are managed and the way they provide care. More American healthcare organizations are stating that they are in a turnaround situation. Although there are no definite guidelines or criteria that place an organization in a turnaround situation, there are certain circumstances that are frequently present. These circumstances most often include a somewhat sudden change from a financially favorable situation. This may include revenue lost and/or increases in cost from operations, decreases in volumes that are most positive and favorable to the organization, sudden changes in key management or board representation, and sudden loss of key managers and staff needed for the successful operation of the organization. Any one of these factors or several can place an organization in such jeopardy that quick and decisive actions must be taken to ensure survival and future success. The specific conditions of a turnaround and the effectiveness of quick and decisive actions are critical for the survival of an organization or a system. PMID- 11924252 TI - The heartbreak of parents patriae. PMID- 11924253 TI - Responses to questions commonly asked about health law. PMID- 11924254 TI - Institutional integrity: values in action. One institution's approach to "grounding" itself in its core values. PMID- 11924255 TI - The Illinois bishops on death and dying. A pastoral letter presents Church teaching in a succinct and understandable manner. PMID- 11924256 TI - The community of concern. An ethical discernment process should include and empower all people relevant to the decision. AB - 1. Always begin by asking explicitly and methodically: Who else would make this a more adequate community of concern? 2. Remember that the need for the community of concern flows from two considerations: the unavoidable presence of constricted consciousness and the nature of moral choice as beneficence--difficult choices concerning human dignity. 3. Pay special attention to the community of concern at the beginning and at key points throughout the process. 4. Define the primary community of concern so that relevant issues include: The decision's impact on the public, and on values, functions, and relationships expertise needed Buy-in and implementation Existing commitments Individuals at the margin who are consistently forgotten 5. Define the secondary community of concern so that relevant issues include: Determining who beyond this primary group will enrich the process With input In implementation In mitigating the harm that results In what ways and how frequently should these individuals/groups be involved? PMID- 11924257 TI - Market segmentation. Sixth in a series examining revenue growth strategies in a difficult health care market. PMID- 11924258 TI - Pastoral care in the LTC setting. Just as death is part of life, spiritual care should be part of long-term care. PMID- 11924259 TI - Recruiting a culturally diverse workforce. Welcoming employees from different cultures is both good business and "good mission". PMID- 11924260 TI - Labor and Catholic health care. The Church's social teaching continues to provide guidance for workplace problems. PMID- 11924261 TI - A values-guided "downsizing". A St. Louis System tried to put its employees' welfare first. PMID- 11924262 TI - A "just wage": more than dollars. Some Catholic executives have drawn up a list of principles concerning compensation. PMID- 11924263 TI - Creating a positive work climate. Ascension Health, St. Louis, bases its human resources policies on Church social teaching. PMID- 11924264 TI - The threat of bioterrorism. PMID- 11924265 TI - Under the big top. PMID- 11924266 TI - When the medium is the message. PMID- 11924267 TI - [Research and scientific progress in 2001]. PMID- 11924268 TI - [Arterial hypertension--latest discoveries. The Danish Society of Hypertension]. PMID- 11924269 TI - [Association between Helicobacter pylori infection and ventricular cancer--more than a suspicion. The Danish Society of Gastroenterology]. PMID- 11924270 TI - [Fibrosis--a nephrological challenge. The Danish Society of Nephrology]. PMID- 11924271 TI - [Treatment of heart arrhythmias. The Danish Cardiologic Society]. PMID- 11924272 TI - [Sepsis--new therapeutic modalities. The Danish Society of Infectious Medicine]. PMID- 11924273 TI - [Progress within pulmonary medicine 2001. The Danish Society of Pulmonary Medicine]. PMID- 11924275 TI - [Clinical pharmacology 2001--important events. The Danish Society of Clinical Pharmacology]. PMID- 11924274 TI - [Late effects after treatment of childhood leukemia. The Danish Society of Pediatrics]. PMID- 11924276 TI - [Monopoly of genetic diagnostics? The Danish Society of Medical Genetics]. PMID- 11924277 TI - [Molecular targeted treatment of hematological diseases with monoclonal antibodies and tyrosine kinase inhibitors. The Danish Society of Hematology]. PMID- 11924278 TI - [Dermatovenereology. The Danish Society of Dermatology]. PMID- 11924279 TI - [Liason psychiatry. The Danish Society of Psychiatry]. PMID- 11924280 TI - [MR urography, a new diagnostic method. The Danish Society of Radiology]. PMID- 11924281 TI - [New computer based therapeutic possibilities in head and neck cancer. IMRT and CAS. The Danish Society of Head and Neck Oncology]. PMID- 11924282 TI - [Significant newer fields within clinical neurophysiology. The Danish Society of Clinical Neurophysiology]. PMID- 11924283 TI - [Scintigraphy combined with CT. The Danish Society of Clinical Physiology and Nuclear Medicine]. PMID- 11924284 TI - [Treatment of varicose veins in Denmark--still place for improvements. The Danish Society of Vascular Surgery]. PMID- 11924285 TI - [Non-prescription postcoital contraceptives. The Danish Society of Obstetrics and Gynecology]. PMID- 11924286 TI - [Challenges of future occupational medicine. The Danish Society of Occupational and Environmental Medicine]. PMID- 11924287 TI - [Research concerning early childhood. The Danish Society of Child and Adolescent Psychiatry]. PMID- 11924288 TI - [Gastroenterological and other research in developing countries. Cooperation between developing and developed countries]. PMID- 11924289 TI - [Concerning the Danish National Committee for Continuing Medical Education and its decision of discontinuation of the subspecialty of clinical neurophysiology]. PMID- 11924290 TI - [Use of cellular telephones and risk of cancer. A Danish cohort study]. AB - INTRODUCTION: The use of cellular telephones has been suggested to increase the risk of cancer. MATERIAL AND METHODS: We carried out a retrospective cohort study of the incidence of cancer in all 420,095 users of cellular telephones during the period of 1982 to 1995. RESULTS: Overall, 3391 cancers were found, with 3825 expected, which yielded a significantly decreased standardised incidence ratio of 0.89. No increased incidence was seen for cancers of the brain or nervous system, of the salivary gland or for leukaemia, cancers which were of a priori interest. DISCUSSION: The results do not support the hypothesis of an association between the use of these telephones and tumours of the brain or salivary gland, leukaemia, or other cancers. PMID- 11924291 TI - [Is use of surgical caps and masks obsolete during percutaneous heart catheterization?]. AB - INTRODUCTION: Despite the lack of scientific evidence, surgical caps and masks are worn routinely by many physicians and nurses in the catheterisation laboratories to avoid local and generalised infections. When we changed our practice, we performed a randomised study to assure the quality of our routine. MATERIAL AND METHODS: All patients undergoing left- or right-sided cardiac catheterisation were randomly allocated to our five laboratories. About two months after the procedure, the patients were sent a questionnaire concerning signs and symptoms of possible inflammation or infection after the procedure. RESULTS: Of 1,034 patients, 855 (82.7%) responded to the questionnaire. Although 25 patients in the caps and mask group vs 19 patients in the other group (6.1% vs 4.3%, ns) had complaints from the procedural access site in the groin, none of these could be ascribed to definite infection. DISCUSSION: The use of caps and masks during percutaneous cardiac catheterisation procedures is based on the concept that the infection rate of patients is reduced. The size of the catheter used, procedure time, and the use of caps and masks by both the surgeon and the assisting staff are discussed. CONCLUSION: The routine use of caps and masks does not seem to have that much beneficial impact on the occurrence of procedure related inflammations or infections in the cardiac catheterisation laboratory. PMID- 11924292 TI - [Secretin treatment of autism?]. PMID- 11924293 TI - [Smoking habits of Danes through 50 years: heavy smokers and non-heavy smokers]. PMID- 11924294 TI - [Methodological problems of questionnaire studies 2]. PMID- 11924295 TI - [Diagnosis of infectious diseases. Brain abscess]. PMID- 11924296 TI - [The editor of Ugeskrift for Laeger]. PMID- 11924297 TI - Two new departments in the Diabetes Educator. PMID- 11924298 TI - The advanced practice diabetes educator. PMID- 11924299 TI - The utility of a portable patient record for improving ongoing diabetes management. PMID- 11924300 TI - Put into practice. Drinking perceptions and management strategies of college students with diabetes. PMID- 11924301 TI - Drinking perceptions and management strategies of college students with diabetes. AB - PURPOSE: The purpose of this research was to explore how college students with diabetes perceive and manage alcohol consumption. METHODS: Fifteen college students with diabetes attending a large northeastern university participated in a single semistructured interview that focused on the impact of starting college on diabetes management and situational obstacles to diet-related self-care. Interviews were audiotaped, transcribed verbatim, and analyzed using the constant comparative method of analysis. RESULTS: Students perceived alcohol as a pervasive part of the university environment and felt strong peer pressure to drink in alcohol-related social situations. Students described 3 distinct drinking practices and identified factors that shaped decisions about drinking. Most students developed personal strategies to manage drinking, usually without guidance. CONCLUSIONS: Some of the students' strategies appeared reasonable for avoiding adverse outcomes of drinking; however, other strategies may have increased their risk of hypoglycemia or poor glucose control. More research is needed to understand how students' management strategies influence diabetes control and to explore how education and counseling from healthcare providers can improve students' management of drinking. PMID- 11924302 TI - Culturally competent dietary education for southern rural African Americans with diabetes. AB - PURPOSE: The purpose of this article is to describe a culturally competent, dietary self-management intervention designed to improve physiological outcomes, diabetes self-management, and costs of care for high-risk African Americans with type 2 diabetes. METHODS: A longitudinal, quasi-experimental design was used to evaluate the effectiveness of the intervention for 23 adult African Americans from a physician practice group in rural South Carolina. The intervention consisted of 4 low-fat dietary education classes, 6 discussion groups, and follow up. Intervention sessions were provided by a dietitian and nurse case managers and framed as social events; families were encouraged to participate. RESULTS: Data suggest that the intervention significantly improved fat-related dietary habits, A1C values, fasting blood glucose, and frequency of acute care visits. A trend in reduction of lipids and weight also was observed. CONCLUSIONS: Culturally competent dietary self-management provides a meaningful approach to focused diabetes education for rural African Americans. Integrating nursing case management provides an innovative method of addressing the more global issues of delivery of care to underserved rural populations and decreasing the high costs of care. PMID- 11924303 TI - Beliefs and attitudes of African Americans with type 2 diabetes toward depression. AB - PURPOSE: This qualitative study was conducted with African Americans with type 2 diabetes to explore beliefs and attitudes about depression. METHODS: Twenty-five adults participated in 4 focus groups. The sessions were recorded, transcribed, and analyzed. The Health Belief Model was used as a theoretical framework for the design and analysis of the focus group data. RESULTS: Five themes pertinent to depression management emerged: (1) There were misconceptions about the etiology of depression and individual vulnerability to depression. (2) Depression was perceived as severe. (3) Treatment was perceived as beneficial. (4) Stigma was a significant barrier to seeking treatment. (5) Cues to action did not appear to change beliefs. CONCLUSIONS: Despite awareness about the severity of depression and benefits of treatment, several barriers and erroneous beliefs may interfere with the ability of African Americans with type 2 diabetes to seek and adhere to treatment for depression. PMID- 11924304 TI - Insulin lispro update. AB - PURPOSE: This paper provides a review of the literature and clinical studies for insulin lispro and updated information on its advantages over regular insulin for various populations of people with diabetes. METHODS: Information was gathered from a search of Medline articles and from review of clinical studies. RESULTS: Patients in various special populations using insulin lispro, with proper adjustment of basal insulin, had a greater reduction in hemoglobin A1C and fewer episodes of hypoglycemia than patients on regular insulin. CONCLUSIONS: More recently published literature shows that due to its faster onset and shorter duration of action, insulin lispro is useful for not only lowering A1C values, but also for reducing hypoglycemic events in various populations with diabetes, including pediatric and pregnant patients, those with gastroparesis, and insulin pump users. PMID- 11924305 TI - Hospital restructuring and downsizing: taking stock: a symposium, Part 1. PMID- 11924306 TI - Hospital reengineering: an evolving management innovation: history, current status and future direction. AB - This article summarizes six years of research on reengineering in hospitals and is the result of two national surveys and eighteen site visits to hospitals that engaged in reengineering in the 1990s. The research shows that actual hospital reengineering differs substantially from that which was initially proposed by early promoters of reengineering. However, this evolved reengineering continues to be implemented by the majority of hospitals in the United States. The authors illustrate how extensive reductions of managers and changes of nursing models have been in the past six years. Data comparing financial and cost competitiveness changes are also shown. The authors then explore the continued experiences of two early proponents of reengineering and find that their competitive outcomes to be in contrast with their early statements. Finally, the authors suggest a number of reasons that may impact on the success or failure of reengineering. PMID- 11924307 TI - Hospital restructuring: does it adversely affect care and outcomes? AB - The past decade has witnessed pronounced changes in the organization of U.S. hospitals, many the direct result of restructuring and reengineering initiative intended to decrease costs and increase productivity. Little is known about how these initiatives have affected clinical care and patient outcomes. Using data from a variety of sources, the authors describe initiatives that hospitals undertook over this period, indicate how staffing changed relative to the case mix of patients receiving care, and examine changes in nursing practice environments over the period from 1996 to 1998. The authors found that apparent increases in nurse-to-patient ratios may be deceiving and that increases in patient acuity and nurses' responsibilities may have increased the workload of nurses in hospitals in ways, when coupled with a deteriorating practice environment, may adversely affect patient outcomes. PMID- 11924308 TI - Managing the consequences of hospital cutbacks: the role of workforce reduction practices. AB - In recent years, hospitals in Canada as elsewhere have witnessed unprecedented downsizing of their workforces. It is generally assumed that planned workforce reductions can have deleterious consequences on an organization's human resources. Scholars and practitioners alike have identified a number of humane or progressive approaches that are widely considered to be effective for organizations undergoing downsizing. This study examines the impact that workforce reduction approaches have on perceptions of organizational performance in a large sample of Canadian hospitals undergoing workforce reductions. PMID- 11924309 TI - Downsizing-initiated job transfer of hospital nurses: how do the job transferees fare? AB - In this longitudinal panel study, the authors compared the reactions to hospital amalgamation of 66 nurses who had been transferred to a different unit for a downsizing-related reason (bumped/displaced, unit closed, redundancy) with the reactions of 181 nurses who remained on their same unit. Prior to any job transfers, the two groups perceived comparable levels of support and held similar attitudes towards their job and the hospital. Two years later, after job transfers had taken place, transferred nurses perceived significantly lower coworker support. They also reported a significantly greater decrease in organizational commitment than nurses who were not transferred. However, both groups reported a significant decrease between time a and time 2 in perceived organizational support, satisfaction with amount of work and career future, hospital identification, and organization trust. Overall, the results indicate that the downsizing associated with the amalgamation of the hospitals had a highly negative effects not only on those nurses who were transferred because of the downsizing but also on those nurses who remained on their original unit. PMID- 11924310 TI - Hospital restructuring: impact on nurses mediated by social support and a perception of challenge. AB - Health care has not escaped the economic rationalization experienced throughout the western world during the 1990s. In Australia, the introduction of Casemix and its accompanying budget cuts has required major restructuring in hospitals. Change can be stressful yet a consultative style, beliefs that change in challenging, and good support were proposed as mediators of the negative effects of change for nurses. This article presents data from 201 nurses working in three Australian hospitals. The results indicate that, despite restructuring changes during the previous 12 months and their high impact upon hospital and nurse conditions, nurses maintained a strong sense of professional efficacy. However, these changes and their impact also predicted nurse burnout. Nurses felt unable to challenge the actual restructuring changes (closed units/beds) however and saw working against the impact of these changes (reduced time, resources) as a challenge. Management's communications about the changes was proposed as a mediator against burnout but this hypothesis was not supported. Nurses considered management to have adopted a top-down approach and this non-consultative style of communication also predicted nurse burnout. The restructuring changes, their impact, and poor communication style, and burnout also predicted nurses' intentions to quit; however, a sense of professionalism mediated this intent. The implications of the findings are discussed. PMID- 11924311 TI - Examining the changing structure of the American health care system: a symposium. PMID- 11924312 TI - Administrators and mergers: a study of administrators' perceptions. AB - Health care mergers have become the organizational strategy of choice in the 20th century to contain costs and to survive economically. Several studies have been conducted on the economic impact of mergers on the organization and on the managerial staff of merged organizations. Areas where there has been limited research are in the realm of the impact of mergers on the employees, the industry, and the public. To address these areas, a study was conducted of the perceptions of health care administrators of both merged and non-merged organizations in the state of Florida. A primary finding of the study was that administrators of merged organizations held a higher opinion of the benefits of mergers in their organizations and in the industry than did administrators in non merged organizations. An unanticipated finding of the impact of mergers was that the increased competition has created a wary, hostile environment for administrators, many of whom refused to participate in this study believing it was being conducted by their competition. The rationale for mergers in health care is based on the market value of competition and future research needs to focus more on the unintended consequences of competition in health care to which this study points. PMID- 11924313 TI - Medicaid managed care and provider consolidation. AB - In the thrust toward constructing economic value, health care provider firms have been consolidating at a marked rate. Medicaid managed care programs have been rapidly emerging with the objectives of containing health care costs and improving services for beneficiaries. However, there are concerns that the trend toward achieving market efficiency through merger is largely incongruent with the economic and health value objectives of Medicaid managed care programs in the states. Discordance among value objectives arises primarily because of inefficient and market concentrating horizontal merger strategies employed by firms and disruptions in quality of care that occur during the transition to integrated health care systems. By promoting vertical integration strategies and filling in the quality gaps created by an active merger environment, Medicaid offices advance state objectives of cost containment and quality while recognizing that providers operate in a complex and competitive environment that necessitates consolidation for organizational survival. PMID- 11924314 TI - Protecting the public interest: issues in contracting managed behavioral health care. AB - In recent years, state governments have begun contracting out behavioral health care to large managed for-profit organizations. This approach to mental health service delivery has resulted in realignment in roles for both state agencies and the managed care organizations. Overnight, state agencies have been transformed into contract managers of multi-million dollar capitated health plans. Managed care companies, in turn, moving into the public-sector market must adjust their service delivery to include key concepts of public interest involvement such as consumer input, public comment, and outreach. This article analyzes three public sector transitions. Common issues in these transformations are used as a springboard for identifying key concerns which states need to monitor when developing managed care contracts. PMID- 11924315 TI - The role of a county health department as a linking-pin organization in a network. AB - This is a case study of a local health department's role in the formation and operation of a network of health care providers. The local health department served as a linking-pin organization in the network and engaged in actions to legitimize itself and the network it formed. Legitimacy management requires the development of pragmatic, moral, and cognitive strategies that can be used for gaining, maintaining or repairing legitimacy. The results of this study indicate a unique government legitimacy management profile in which there is a predominant use of cognitive and pragmatic actions intended to gain legitimacy. The implications of this profile are discussed. PMID- 11924316 TI - Strategic management and health workforce policy. AB - Among the many consequences of health care restructuring is the impact such changes have on the training requirements for the health professions. Since workforce planning has been difficult and sometimes controversial in relatively stable times, it is likely to be even more problematic amid the turbulent changes ahead as the U.S. health care system restructures for the 21 century. Strategic management models emphasizing stakeholder involvement offer a middle ground between the extremes of government mandates and free markets by engaging a variety of participants with a stake in the planning outcome. The following report on the New Jersey effort to engage a variety of health care stakeholders in a participatory management process to shape the state physician workforce may provide useful insights for both managers and policy-makers. PMID- 11924317 TI - [Conflictual opinions on the future of nursing education: search for consensus has consensus]. PMID- 11924318 TI - [Otorhinolaryngology--V. Hemorrhage of the ENT area requires rapid management]. PMID- 11924319 TI - [German Organ Transplantation Foundation: partner of the hospital]. PMID- 11924320 TI - [Organ donation, procurement and transplantation: a responsibility for life]. PMID- 11924321 TI - [Nursing care of a potential organ donor: brain death... and now what?]. PMID- 11924322 TI - [Deciding on organ donation: when fate overtakes reality]. PMID- 11924323 TI - [Work in a transplantation center of the Stuttgart clinic: before organ arrangements the calculations are done]. PMID- 11924324 TI - [Nursing a patient with kidney transplantation: characterized by a changed life style]. PMID- 11924325 TI - [Special nursing processes after orthotopic heart transplantation: the goal is independent living]. PMID- 11924326 TI - [Transplantation Nursing Professional Circle: a platform for nurses]. PMID- 11924327 TI - [Learning in nursing: problem based learning]. PMID- 11924328 TI - [Precision in intensive care: the unit with the greatest requirements]. PMID- 11924329 TI - [Stress--causes, reactions, management: everyone experiences it, no one likes it]. PMID- 11924330 TI - [Terminal care nursing in Australia: experiences at the other end of the world]. PMID- 11924331 TI - Clinical practice guidelines. USPSTF recommendations. AB - The set of USPSTF recommendations provides a valuable resource for clinicians in varied practice settings. Although the 2nd edition of recommendations addressed over 200 topics, only six statements have been released as part of the 3rd edition thus far. Others, not reviewed in this article include: screening for newborn hearing, screening for bacterial vaginosis during pregnancy, and screening for lipid disorders. The recommendations are conservative and, like all clinical practice guidelines, should be considered in light of one's philosophy of practice. The recommendations meet many of the criteria addressed in the initial column of this series. The factors considered by the panel members are identified, including accuracy of available screening methodologies, cost factors, and benefit/risk determinations. The evidence on which recommendations are based is rated. Exceptions to the recommendations are identified. For instance, the authors of the paper on skin cancer screenings indicate that they did not consider studies based on persons with familial skin cancers. Although no recommendation is made for or against skin cancer screenings, ways in which the skin can be assessed during physical examinations performed for other reasons are identified. The importance of individual patient preference is included. The USPSTF panel is multi-disciplinary and includes [table: see text] (http://www.guideline.gov). The statement on aspirin therapy was published in the Annals of Internal Medicine and is available, along with a second article discussing the evidence in more depth (Hayden, Pignone, Phillips, & Mulrow, 2002), at: http://www.annals.org. Print versions are available through the Agency for Healthcare Research and Quality (AHRQ) Publications Clearinghouse (1-800-358 9295). Readers are encouraged to obtain full copies of the recommendations that are applicable to their practice as they become available and to assess their potential application in practice. PMID- 11924332 TI - Diagnosing and treating Alzheimer's disease: a practitioner's overview. AB - PURPOSE: To provide an overview of current diagnostic protocols of Alzheimer's disease (AD), screening techniques, tests, and a review of standard and new treatments. DATA SOURCES: Selected articles from the scientific literature, online sources, and standard texts were examined. CONCLUSIONS: Standard tools, such as the Geriatric Depression Scale, the Mini Mental State Evaluation, and the 7-Minute Screen, are useful in the primary care setting for screening elderly patients. Other tools, such as the Clinical Dementia Rating (CDR) and the Global Deterioration Scale, are useful for staging and monitoring progression of disease and response to treatment. Clinical diagnostic testing is still in developmental stages, but the hope is to have a reliable and objective diagnostic tool in order to diagnose AD in the earliest stages. IMPLICATIONS FOR PRACTICE: The diagnosis and treatment of AD is becoming a more frequent challenge in the primary care office. Clinicians must keep abreast of the rapid changes in new technologies in order to make informed diagnostic and therapeutic decisions. PMID- 11924333 TI - The primary care nurse practitioner: advocate for self care. AB - PURPOSE: To critically analyze the role of the primary care nurse practitioner (NP) as advocate in promoting health through self care. DATA SOURCES: Selected philosophical and theoretical texts and articles, scientific literature, professional standards, and ethical arguments. CONCLUSIONS: Philosophical reasoning and nursing theory must be used to validate and direct human practice. If self-care and well-being are desirable as an end and are defined as a movement toward self-determination, then the philosophical goal of the NP must be the protection and promotion of self care and autonomy through client advocacy. IMPLICATIONS FOR PRACTICE: The role of the NP must be defined by its philosophy of care rather than by its function. The influence of philosophical reasoning, nursing, and critical social theory impact the ability of the NP to promote self care through advocacy. Understanding the lived experience is an essential component of advocacy. PMID- 11924334 TI - The master's portfolio: validating a career in advanced practice nursing. AB - PURPOSE: To describe the process of portfolio development and faculty review that offers graduate credit within a framework of structural empowerment and mentoring for documented educational and clinical accomplishments for family nurse practitioners (FNPs). DATA SOURCES: Selected literature and examples from one student's portfolio to illustrate the process. CONCLUSIONS: The degree granting process assists certified FNPs to fulfill career aspirations through achieving graduate level education at the University of Massachusetts, Amherst, School of Nursing. IMPLICATIONS FOR PRACTICE: The portfolio process affords practicing FNPs not only the opportunity to acquire an advanced degree but a framework for examining and validating a career. For the student in this exemplar, it was a "significant, introspective, satisfying learning experience," a rare opportunity in a busy professional life. PMID- 11924335 TI - The role of exercise in the African-American woman with type 2 diabetes mellitus: application of the health belief model. AB - PURPOSE: To determine whether aging African-American women with a diagnosis of type 2 diabetes mellitus who maintain a regular exercise regimen possess different health beliefs and benefit from greater glycemic control than those who do not exercise regularly. DATA SOURCES: A 32-item health belief model diabetes scale was administered to a convenience sample of 31 African-American women with type 2 diabetes. CONCLUSIONS: A group t-test identified statistically significant differences between "exercisers" and "non-exercisers" in perceived benefits and barriers to exercise and glycemic control. IMPLICATIONS FOR PRACTICE: Findings may be utilized in the development of diabetic educational programs and compliance-enhancing treatment interventions that focus on the barriers and benefits of diabetes self-management. PMID- 11924336 TI - The relationship of actual height loss with health-seeking behaviors and risk factors in perimenopausal and menopausal women. AB - PURPOSE: To explore whether there is a difference in the perceived height and actual height in the perimenopausal or menopausal woman and discover the difference's effect on osteoporosis health-seeking preventive behaviors and risk factor awareness. DATA SOURCES: Sixty-three perimenopausal and menopausal women, aged 45-70 years, who presented at three Midwestern clinics for their annual physical exam. Data were collected with The Osteoporosis Questionnaire, which included the Osteoporosis Risk Questionnaire, Health-O-Meter height measuring stick, tape measure with inch-rule for arm span measurements, and balance scale for weight. CONCLUSIONS: There were a significant relationship between actual height loss and osteoporosis risk factors (r = 0.41595, p = 0.0007) and a trend for a relationship between adjusted height loss and osteoporosis risk factors (r = 0.2407, p = 0.0574). IMPLICATIONS FOR PRACTICE: Due to the great expense of current testing for bone mass density loss, the results of this study may help clinicians more readily identify markers for increased risk of osteoporosis through simple height measurements and osteoporosis risk factor assessments during annual visits. PMID- 11924337 TI - Challenges of outcomes research for nurse practitioners. AB - PURPOSE: To describe the numerous methodological challenges nurse practitioners (NPs) face in designing and conducting outcomes research and provide practical tips to implement an outcomes study within an institution. DATA SOURCES: Review of world wide scientific literature on outcomes research. CONCLUSIONS: Nurse practitioners must be aware of the challenges of conducting outcomes research. Challenges associated with variable definition, designing outcomes studies, use of data sets, and instrument development and selection must be understood prior to undertaking an outcome study. IMPLICATIONS FOR PRACTICE: Valuable studies have laid a foundation for evidence of quality care provision by NPs. Now is the time to measure patient outcomes of the NP care longitudinally over significant periods of time. PMID- 11924338 TI - California releases proposed nurse-to-patient ratios for acute care hospitals. PMID- 11924339 TI - Rural and public health: connecting vision, values, and technology. PMID- 11924340 TI - Safe nursing and patient care act of 2001. PMID- 11924341 TI - National nursing legislation updates. PMID- 11924342 TI - Time after time. Mandatory overtime in the U.S. economy. PMID- 11924343 TI - The Verona Benchmark: applying evidence to improve the quality of partnership. PMID- 11924344 TI - Benchmarking to promote better health. PMID- 11924345 TI - Benchmarking for investment for health. PMID- 11924346 TI - Promoting health in Wales--strengthening partnerships for investment for health. PMID- 11924347 TI - Using the Verona Benchmark to introduce health promotion concepts and the investment for health approach in the city of Nova Gorica, Slovenia. PMID- 11924348 TI - Principles, methodology and practices of investment for health. PMID- 11924349 TI - Investment for health in an old mining industry area of Sweden. PMID- 11924350 TI - The Verona Initiative: the process of developing partnerships at local level. PMID- 11924351 TI - Recovering maxillofacial trauma patients: the hidden problems. AB - This longitudinal study highlights the psychological and functional problems that can result from maxillofacial trauma. This is the first study to report outcome at one year. A total of 147 patients admitted for surgery following facial trauma were recruited over a seven-month period. Three questionnaires were used to record patient-derived levels of dysfunction: the Hospital Anxiety Depression scale, a modified University of Washington Quality of Life questionnaire and five non-validated facial trauma items. At one year 46 patients (31%) responded. Although there were significant improvements in scores from pre-operatively to one year, with all patients being discharged from outpatient follow-up, there was a substantial level of subjective symptomatology. Most notable was the level of anxiety and depression, which were present in 30% of the sample at both time points. Health-care professionals tend to underestimate the long-term effects of maxillofacial trauma. To improve patient care, greater appreciation of these problems is required at the time of initial management. PMID- 11924352 TI - Do we need to reappraise our method of interpreting the ankle brachial pressure index? AB - This prospective study aimed to investigate if there is a relationship between ankle brachial pressure index (ABPI) and brachial systolic pressure when measured using Doppler ultrasound. Sixty-two limbs (36 patients) in which arterial disease and diabetes mellitus could be excluded using published blood pressure and flow criteria were included in the study. All measurements were made with a Scimed PVL 50 (Bristol UK) using a revised version of the Whiston method and a one-minute treadmill walk. Statistical analysis was carried out using Pearson's product moment correlation and the t test. A relationship was found between the ABPI and brachial systolic pressure, with statistical analysis indicating that the ABPI was highest in hypotensive patients and lowest in hypertensive patients (r = 0.385, p < 0.01). This may suggest that the current practice of assigning one particular ABPI value as normal (1.00) is not reliable and that the way in which the index is interpreted needs to be reappraised. PMID- 11924353 TI - Malignant wound management: what dressings do nurses use? AB - This paper reports on the findings from the first part of a three-phase project that aimed to identify nursing strategies used in the management of malignant wounds. The difficulties relating to the management of these wounds and the significant physical and psychological impact on patients are described. A quantitative postal survey aiming to identify the types of dressing used in the care of malignant wounds was sent to specialist nurses working in oncology and palliative care in New South Wales, Australia. Additional qualitative data showed that the major issues were coping with odour and meeting the financial costs of the dressing products. The long list of products compiled for this research demonstrates the complexities nurses face when selecting dressings for the management of malignant wounds. Furthermore, there are no clear recommendations to guide nursing practice. This study provides a framework for subsequent phases of the project and will hopefully lead to the development of guidelines for best practice in malignant wound management. PMID- 11924354 TI - A difficult wound to treat? PMID- 11924355 TI - Klippel-Trenaunay syndrome: what are the implications for wound care? AB - Patients with Klippel-Trenaunay syndrome may present with problems such as allergic contact dermatitis and ulceration. This case study explains how it should be diagnosed and treated. PMID- 11924356 TI - Producing criteria for static mattress tender and purchase. AB - When selecting mattresses criteria such as health and safety plus clinical and cost-effectiveness should be considered. These can then form the basis of contract tender specifications. PMID- 11924358 TI - Nutrition and wound healing: what is the link? AB - Given the important role played by nutrients in wound healing, all patients with wounds should receive a nutritional assessment. Action must then be taken to address any deficiency. PMID- 11924357 TI - Exploring the use of an alginate dressing for diabetic foot ulcers. AB - Few studies have investigated the use of dressings on patients with diabetic foot ulceration. This paper reports the results of a non-comparative, two-centre study investigating the performance characteristics of SeaSorb dressing in patients with this condition. This was an exploratory study to determine the potential use of this dressing, and not a large randomised control trial. All patients had diabetic foot ulceration of 1 cm or more in diameter, classified as Wagner grade I or II, with an ankle brachial pressure index of > 0.4. The investigators carried out objective assessments including photography and ulcer tracing as well as subjective assessments of exudate levels, wound bed condition, peri-ulcer skin condition, ease of application and removal of the dressing, dressing conformability, discomfort during dressing application and removal, and clinical signs of infection. Investigators recruited 41 patients in two sites (31 in site one and 10 in site two). Of these, the findings relating to 39 patients could be evaluated. The patients were treated for a maximum of six weeks or until the ulcer healed. The study found that 28.2% (11/39) of the ulcers healed within the six-week period. Overall, there was a significant reduction in mean ulcer area from 2.8 cm 2 to 1.02 cm 2 from week 0 to week six. The relative ulcer area showed a significant decrease from 100% to 33%. For patients who experienced ulcer pain (11 patients), its intensity decreased over the six weeks. Severe maceration was not reported for any patient throughout the study. Six patients required treatment for infection. A total of 12 adverse events were reported: seven mild to moderate and five severe. None were directly attributed to the study dressing. PMID- 11924359 TI - [Harmony of the facial profile. Part IV. Sagittal rules]. AB - Different inclines characterize the respective parts of the profile. These inclines are identified by the values of the angles created by the individual reference line and the lines representing the inclines. It is shown how these inclines should relate to achieve individual formal perfection of the profile. PMID- 11924360 TI - [Dental treatment and socioeconomic status in the period of 1981-1995]. AB - Between 1981 and 1995 the number of people in the Netherlands with a complete set of dentures fell by an average of 0.8 per cent per year. This means that the number of Dutch inhabitants with complete dentures dropped from 3.5 million in 1981 to 2.5 million in 1995. In general, relatively more people with a lower than with a higher socio-economic status have complete dentures. These socio-economic differences did not change in the period 1981-1995. In the same period, the number of consultations at which the dentist provided preventive treatment increased by 1 per cent per year. The results do not show any important differences in preventive aid between socio-economic groups. Changes in the reimbursement system for dental treatment, introduced on 1 January 1995, have led to a decrease of the number of dental consultations by people on a low income with non-private health insurance. PMID- 11924361 TI - [Dental research in the Netherlands. Citations in publications]. AB - This study examines the citation frequency (via the Science Citation Index) of publications by Dutch dental scientists that have appeared in the international literature since World War II. It was found that most citations were obtained by a relatively small group of researchers, many of them working in the fields of cariology and periodontology. High scores were particularly found among scientists who had their academic training in disciplines other than dentistry. Out of 109 frequently cited papers over 25% was microbiology-oriented and most of them could be characterized as original basic dental research. PMID- 11924362 TI - ['No space for the freeway space']. PMID- 11924363 TI - [Salivary tests for caries and erosion risk factors]. AB - Apart from a health questionnaire and oral health examination with a clinical history, it is also important to inquire after oral and salivary complaints. Supplementary it is useful to assess dietary habits in patients at risk and to perform a number of simple salivary tests. Interpretation of all data obtained through these investigations, will enable the identification of a number of caries and erosion risk factors. On the basis of these risk factors a preventive treatment plan can be made for the patient. These data provide the possibility to evaluate the success of the preventive treatment plan after a few years and to make adjustments, if necessary. PMID- 11924364 TI - [Sleep apnea. Some snoring people also have trouble during the day]. PMID- 11924365 TI - [Trends in dental caries prevalence amongst schoolchildren in the Hague. A comparison with the results from a survey in 1996 and previous years]. AB - Starting in 1969, periodic cross-sectional examinations of schoolchildren have been carried out in the city of The Hague (the Netherlands). The results of 1996 show that no further change in caries prevalence has taken place in 5- and 7-year old Dutch children of low, medium and high SES. In 11-year-old children of medium and high SES a further decrease in caries prevalence could be observed in 1996. Between 1989 and 1996 a significant caries decrease in the deciduous dentition was found amongst 5-year-old children from Turkey and Morocco. No improvement of dental health in 7-year-olds was found. Since 1989 an increase in percentages of caries-free 11-year-olds from Turkey took place whereas this percentage in children from Morocco tended to stabilize (26 and 29% respectively in 1989 and 1996). PMID- 11924366 TI - [Dentistry for children. Introduction]. PMID- 11924367 TI - [Unhealthy differences. Part I. Socioeconomic deprivation and health]. AB - Socioeconomic deprivation often concurs with deprivation in health. In this paper, the available evidence on socioeconomic health differences in The Netherlands is summarized. Furthermore explanations for the existence of these differences are given. Socioeconomic deprivation often leads to a worse health, but the reverse can also occur sometimes. Policies on the causes of these health differences usually concern fields like income distribution and employment. Health care can offer a compensation for the adverse health effects of socioeconomic deprivation. PMID- 11924368 TI - [Unhealthy differences. Part II. Dental aspects]. AB - In the seventies dental caries was present in almost all Dutch children. Nowadays the child population can be divided into two groups: children with no dental caries and children with a relatively high level of dental caries. The lowest percentage of caries free children is found in low SES (= Socio-economic Status) groups. Within the low SES group children of Turkish of Moroccan origin have a higher caries prevalence compared to the youngsters of Dutch origin. However, differences in the level of caries prevalence between children of Dutch and Turkish or Moroccan origin in recent years have declined. The level of oral health in children from risk groups seems to be due to irregular toothbrushing and consequently irregular use of fluoride toothpaste. PMID- 11924369 TI - [Caries treatment strategy. 'Tip the balance'--drilling or not drilling]. AB - Traditionally caries treatment strategy has been based on the detection of caries lesions, and directed to a restorative procedure whereby the detected defects were filled. However, the detection of lesions is not synonymous to caries diagnosis. The main purpose of caries diagnosis should be the determination of the caries activity. The diagnosis is based on several parameters, which are evaluated by an intellectual process in a dentally trained mind. The treatment should be confined, in the first instance, to stopping the caries process, and to tipping the balance in the right direction, so that the caries can cure or anyway is prevented from extending. The treatment philosophy has to be pointed to a general approach of the disease caries in the patient who it concerns on the one side, and to a local approach of individual lesions on the other hand. In the second case a lesion can be treated in a symptomatic, invasive manner or, even better, by causal, not-invasive treatment. PMID- 11924370 TI - [Prevention of caries in children]. AB - There is a stagnation in the previously ongoing improvement of the dental health of children. Therefore, an evaluation of the guidelines for dental care is necessary. Prevention should be based on an improvement of home care. It is advised to use a dentifrice with 500 to 750 ppm fluoride till the age of five. Professional applications in the dental office are most beneficial when early signs of decay are observed. Indications for professional applications are proposed. PMID- 11924371 TI - ['Hidden caries' and radiographic studies in children's teeth]. AB - Proximal cavities in deciduous molars are often clinically detected at the time the pulp is involved in the caries process. Besides the early detection of lesions in deciduous teeth in children with a high caries-risk the bite-wing also plays a role in the detection of occlusal 'hidden caries' in non-risk children. In case of 'hidden caries' curative intervention is indicated. Guidelines for the use of bite-wing radiograph's in children are discussed. PMID- 11924372 TI - [Dental restoration materials in pediatric dentistry]. AB - Restorative materials in pediatric dentistry have to fulfill special requirements. They should be easy to handle and applicable in a not always dry mouth. They should potentially be adhesive in order to avoid too much mechanical preparation. They do not have to be extremely wear resistant as the dwell time of the restorations is relatively short. Glass-ionomer cements and in particular the resin modified types possess properties which make them almost ideal for the required purpose. PMID- 11924373 TI - [Periodontitis in children]. AB - In children with a healthy periodontal condition the pocket depth may vary between 1 to 2 mm. In the mixed dentition it may increase up to 3 mm and in the permanent dentition the pocket depth usually decreases again to between 1 and 2 mm. If a pocket of 5 mm or more is diagnosed one should be aware of periodontal breakdown, i.e. periodontitis. Periodontitis is usually defined on the basis of age, localisation and the presence of systemic diseases. Frequent periodontal probing is necessary to detect periodontal breakdown in an early stage and enables subsequent periodontal treatment. Involvement of the parents in the treatment is important. Their major contribution in the treatment is daily careful cleaning of the dentition of the child. PMID- 11924374 TI - [Amelogenesis imperfecta in young patients]. AB - Amelogenesis imperfecta is a genetic disturbance in the formation of enamel. The condition can be classified as a hypoplastic type and a hypomineralized type. Both types are normally very inconvenient for the patient and treatment should be started as soon as possible. Dental treatment should aim at maintaining the height of the bite and improvement of occlusal function and esthetics. In a few case presentations the possibilities and limitations of adhesive composite restorations in young patients with amelogenesis imperfecta are discussed. PMID- 11924375 TI - [Cleft lip and palate: thoughts about associated anomalies]. AB - During two years all new patients referred to the cleft palate team in Nijmegen, were examined by a clinical dentist, to evaluate the occurrence of associated anomalies or syndromes. In 33% of the patients with all types of clefts associated anomalies were found. A percentage of 56% was found for cleft palate only, and of 14% for cleft lip (palate). It is important to make a classifying diagnosis of these associated anomalies because they may have important implications for recurrence risks and prognosis. The percentage of associated anomalies is the highest in the group of patients with cleft palate only or submucous clefts. Therefore, the dentists have to be aware of the presence of submucous clefts and their consequences. PMID- 11924376 TI - [Dentition without end]. PMID- 11924377 TI - [The influence of scientific publications on the general dental practice]. PMID- 11924378 TI - [Fall congress NVT: 'ethics in the practice']. PMID- 11924379 TI - [From reparation to regeneration: devil's work or God's wonder?]. PMID- 11924380 TI - [Plaque: full blood vessels, hollow jaws]. PMID- 11924381 TI - [Oral pathology in children. Part II. Abnormalities of the teeth]. AB - An overview is presented of the various abnormalities of the teeth in children. Caries and periodontal diseases have not been discussed since each dentist is familiar with these diseases. Abnormalities of the teeth may occur as part of various syndromes. Occasionally, the dental findings lead to the detection of such underlying syndrome. PMID- 11924382 TI - [Amalgam use and mercury emission in the Netherlands]. AB - An overview is presented of the emission of mercury to the environment by restoring teeth with amalgam, extraction, replacement of amalgam, and the final phase of teeth. Important input data were the trade figures of amalgam. The emission of mercury from amalgam fillings are characterized by diffuse spreading. Many small sources are together a substantial source of the leakage of mercury to the environment. Amalgam adhering to cotton-wool and in extracted teeth deliver a relevant contribution to the mercury load in municipal waste. The total mercury emission from dental amalgam to the environment in the Netherlands is at the estimate 500 kg a year, with a worst case maximum of 935 kg. PMID- 11924383 TI - [Practice guidelines in dentistry. An insight into the Dutch situation]. AB - Practice guidelines provide opportunities for monitoring and improving dental health care. A survey was conducted among 78 departments at dental societies, scientific dental associations and health care insurance companies in the Netherlands to obtain insight into the number and quality of practice guidelines available. The response rate was 67%. Written materials were analysed and 21 practice guidelines were identified as such. The results of the analysis indicate that many initiatives to construct and implement practice guidelines have been taken. However, these initiatives were insufficiently coordinated and existing guidelines may be subject to improvement. Main improvement would be to demonstrate the validity and reliability of the existing guidelines and, more in general, to provide a sound scientific basis to the development and implementation of practice guidelines. PMID- 11924384 TI - [Tooth wear. Classification and terminology]. AB - Tooth wear rarely occurs as a result of one factor alone. Non-carious loss of tooth tissue can be subdivided into attrition, demastication, abrasion, fractionation, and erosion. Clinically, the tooth tissue loss may result from any combination of these types of wear. The first essential principle in the management of tooth wear is, as far as possible, to remove the cause. Secondly, restoration may be indicated. PMID- 11924385 TI - [Gallium: an alternative for amalgam?]. AB - Some of the physical properties of gallium-based dental alloys for restorative goals equal those of dental silver amalgam, but the results of the relatively few and short-term studies of their clinical behaviour differ. Corrosion, discoloration, rough surface, expansion and fractures of the margins and possibly the teeth, may pose serious problems and are assessed to make the clinical use premature. Handling characteristics are less favourable than for silver amalgam. Gallium alloys release a substantial amount of gallium. Results of toxicity studies with cell cultures differ. Compared to some brands of silver amalgams the alloys seem to be somewhat more cytotoxic during a longer period of time. However, the gallium released by the restorations would be insufficient to harm the patients. Implants of (early) alloys evoked more serious reactions than silver amalgam. Sensitization is not reported, but few restorations have been made and allergic studies are scarce. PMID- 11924386 TI - [Oral pathology in children. Part III. Abnormalities of the jaw bones]. AB - Diseases of the jaw bones in children can be monostotic or polyostotic. Examples are fibrous dysplasia and Langerhans cell granulomatosis. Sarcomas of the jaw bones are rare, but do occur with some preference in children. The dentist should be aware of the clinical and even more so of the rather typical radiologic changes in such diseases. PMID- 11924387 TI - [Beware of pitfalls. Problems in the somatization patient]. AB - Somatization is a process that makes that psychological and social problems are experienced physically and presented as a physical complaint. This phenomenon should be considered as a potentially dangerous pitfall for the dentist. Not recognizing the underlying problems may lead of unnecessary and inadequate treatments. The purpose of this article is to provide the dentist with practical guidelines for proper assessment, treatment and referral of this patient category. PMID- 11924388 TI - [Problems with dental information given to patients. A report by the consumer organisation]. AB - The objective of this study was to investigate the information problems of Dutch dental patients in the years 1993-1995. Using a self-developed scale, the dental files of a consumer organisation were analysed and the information problems counted and categorised. Of all the dental dossiers 48% concerned information problems. The largest categories were: insufficient information about finances, insufficient information about the treatment/outcomes and the negligence of the information, complaints and questions of the patients. Together these categories make up 75% of the information problems. The results of this study stress that dentists should offer better structured information. More attention should also be paid to the abilities of the dentists to be communicative and to manage conflicts. PMID- 11924389 TI - [Jan Willem: a special case?]. PMID- 11924390 TI - [Focal infection: new points of view]. PMID- 11924391 TI - [Series on masticatory muscles--introduction]. PMID- 11924392 TI - [Masticatory muscles. Part I. Functional anatomy of the masticatory muscles]. AB - The anatomical design of the jaw-closing and jaw-opening muscles is of importance for their capability to produce forces and displacements. The length of the sarcomeres is a major determinant for both force and shortening range. The maximal work, force and shortening range each muscle is capable of producing, are proportional to the architectural parameters volume, physiological cross sectional area and fiber length, respectively. Compared to the jaw openers, the jaw closers are characterized by shorter sarcomeres at the closed jaw, larger masses of contractile and tendinous tissue, larger physiological cross-sectional areas, shorter fibers and shorter moment arms. In addition, architectural features vary across the muscles of the same functional group and in some muscles significant intramuscular differences are found. In general, the jaw closing muscles have architectural features that suit them for force production, whereas the jaw-opening muscles are better designed to produce velocity and displacement. PMID- 11924393 TI - [Supplementary insurance: yes or no? An investigation into the motives of patients]. AB - Of the group of dentate adults taking part in the Public Health insurance scheme, 78% has opted for a supplementary insurance for dental care after the dental health care reform on January 1, 1995. For edentulous people this figure is 33%. Both for people who choose for a supplementary insurance and for those who did not, financial motives were most important. For the dentates visiting a dentist regularly, the time period between two check-ups has grown from 6.0 months to 6.9 months. The extent to which check-ups are postponed is not different for people with or without a supplementary insurance for dental care. PMID- 11924394 TI - [Temporomandibular joint cracking: background and diagnosis]. AB - Although patients often report painless clicking of their temporomandibular joint, treatment is not required. However, sometimes a clicking joint leads to a painful limitation of the movements of this joint. Why this happens is unknown. It is therefore recommended to monitor the clicking joint over time. Opto electronic recording of condylar movements might offer additional possibilities in the assessment of clicking joints. PMID- 11924395 TI - [Changes in secretion and composition of saliva with aging]. AB - About 30% of the elderly suffer from oral dryness and related complaints. Therefore, it is often assumed that salivary secretion reduces with age. Nevertheless, salivary flow rate data in healthy elderly--that are persons without any medication and not suffering from any (treatment related) salivary gland disorder--revealed no significant age-related decrease other than a slight decrease of the secretion from the (sero)mucous glands under conditions of minimal or extended stimulation. No age-related changes were observed in the secretion of parotis saliva. In contrast, some age-related changes occur in the organic composition of saliva. The reported decrease in the concentrations of kallikrein, sIgA, low-molecular mucin and high-molecular mucin might be related to a somewhat decreased immunologic and non-immunologic defense of the oral cavity in the elderly. PMID- 11924396 TI - [An osteoma, to treat or not to treat?]. PMID- 11924397 TI - [Health globalization. I]. PMID- 11924398 TI - [Theme: implants in the upper jaw]. PMID- 11924399 TI - [Success of implants in the moderately resorbed edentate maxilla]. AB - In applications of implants in the moderately resorbed maxilla factors such as bone quantity and the loading of the bone are of importance for the prognosis. Long-term studies have shown that for the maxilla the results were less good than for the mandible. The main reason for this is the condition of the bone, especially reduced bone quantity together with a thin corticalis an low bone density. Beside these factors there are also the general negative factors such as rheumatoid arthritis, osteoporosis, smoking and radiotherapy. The prognosis of implant supported prostheses is not primarily determined by the number of implants, but by the condition of the bone and to some extent by the splinting effect of the implants. PMID- 11924400 TI - [Implants for the extremely resorbed edentulous maxilla]. AB - The success rate of implants and implant-supported superstructures in the (extremely) resorbed edentulous mandible has as of yet not been equalled by similar treatment options carried out in the resorbed maxilla. The pre-treatment phase (diagnostics, indication and treatment planning), if carried out mutually by the prosthodontist and the implantologist, is of great importance if one wants to obtain high standard treatment goals. To ensure an optimal retention and support of the superstructure and loading of the implants, the insertion of preferably a minimum of six implants, evenly distributed over the maxilla is necessary. PMID- 11924401 TI - [Classic method of implantation in the upper jaw]. PMID- 11924402 TI - [Technical problems with superstructures for the maxilla]. AB - The dental lab is an important factor in treating patients with severely resorbed maxillae with implants, but it still occurs that the dental technician is not or not enough involved in the treatment planning of these patients. In this article a number of frequently occurring problems is discussed, especially the number and the location of implants, the space for the superstructure and the planning of the costs. Some suggestions for improvements are proposed. PMID- 11924403 TI - [Pre-implantation surgery of the atrophied maxilla. A review of the literature]. AB - The aim of preprosthetic reconstructive surgery of the resorbed maxilla is to create sufficient bone of good quality for implant placement. The classification of the edentulous jaws by Cawood and Howell is mostly used as point of departure in planning preprosthetic reconstructive surgery. In this review article some procedures and results are mentioned as described in recent literature. PMID- 11924404 TI - [Splitting and widening of a narrow jaw ridge in the edentulous maxilla]. AB - One of the most frequently occurring problems in oral implantology, especially in the maxilla, is the reduction of the alveolar ridge, caused by bone resorption in the edentulous and partly edentulous maxilla. A usually successful but clinically sometimes complicated way of solving this problem is the augmentation of the resorbed parts of residual ridges, particularly when autologous bone is being used. Implant placement usually follows in a later stage. The bone-splitting and bone-widening technique, on the other hand, is a more obvious method for the immediate placement of implants in those cases where the dimensions of the residual ridge are reduced by only some degree. When the resorption of the residual ridge is extreme, this method can eventually be combined with guided tissue regeneration. Crestosplit instruments are specially developed osteotomes for cleaving and widening the crest of the residual ridge, whilst, at the same time, compressing the cancellous bone. In applying this procedure, a bone preparation can be made with dimensions very close to the diameter of the implant to be placed, and without losing important bone-parts as in the case of drilling. Upon the completion of this procedure, only the last drill with the dimensions of the implant has to be used and the implant can thus be placed in the same session. In edentulous cases where the resorption is almost nil, bone preparation with the aid of this method is also to be preferred as the compression of the corticocancellous bone will considerably increase the bone contact with the implant surface. Especially in the maxilla this will lead to a better prognosis of the survival rate of the implant and to better esthetic results of the final prosthetic restoration. PMID- 11924405 TI - [Surgical correction of the intermaxillary relation for placement of implants]. AB - The loss of teeth is associated with alveolar bone resorption. Severe resorption of the maxillary alveolar process may cause persistent instability and loss of retention of the upper denture. Large intermaxillary discrepancies may complicate the treatment with implants to retain maxillary overdentures. Reconstructive surgical treatment methods that create sufficient bone volume for the placement of implants and at the same time normalize intermaxillary relationships, provide the basis for adequate oral rehabilitation. PMID- 11924406 TI - [Complications of the surgical treatment of the severely resorbed edentulous maxilla]. AB - Complications during reconstructive surgery for placement of endosseous implants in the severely resorbed maxilla are described, based on recent literature. Detailed long term follow-up studies on complications are lacking. Clinical experience has shown that this type of surgery is prone to complications. These complications must be taken into account when indicating and planning this type of surgery. PMID- 11924407 TI - [Augmentation of the maxillary sinus floor and alveolar ridge for placement of endosseous implants in the edentulous maxilla]. AB - Placement of endosseous implants in the atrophic maxilla is often restricted because of lack of supporting bone limiting placement of implants of adequate length in a prosthodontically optimal position. There are several surgical procedures to create sufficient volume of bone for the placement of implants. In this paper, a technique is described for augmentation of the maxillary sinus floor and simultaneous widening of the alveolar crest with autogenous bone. PMID- 11924408 TI - [Bone quality after sinus floor augmentation]. AB - The problem of insufficient alveolar bone in the edentulous maxilla caused by resorption and pneumatization can be overcome by augmentation of the sinus floor to increase bone volume for the placement of dental implants. The quality of bone which is achieved after sinus floor augmentation is hardly known. This study describes the histologic results obtained three till six months after sinus floor augmentation with autogenous bone from the iliac crest in patients with severe maxillary bone resorption. The bone biopsies taken from the implant sites showed a substantial bone volume with a mature trabecular pattern and active bone growth. It was concluded that with the obtained bone quality, the sinus floor augmentation procedure can be a good treatment modality for the rehabilitation with implants in patients with severe maxillary bone atrophy. PMID- 11924409 TI - [Five-year evaluation of implants in the resorbed maxilla]. AB - The survival of implants placed in the resorbed maxillae was investigated. Both edentulous and partially edentulous patients were evaluated including those who underwent 'sinus lift' procedures. The group of patients with a sinus floor augmentation showed a five-year cumulative survival rate varying between 100% for fixed bridges on implants in the partially edentulous maxillae and 75.6% for implants under overdentures placed in severely resorbed edentulous maxillae. It is concluded that placement of implants in the augmented sinus-floor is justified, if the patient is well informed. The procedure can provide a good solution for the prosthetic problems of patients with a resorbed maxilla. PMID- 11924410 TI - [A prosthodontic concept using implants in the resorbed edentulous maxilla]. AB - For the treatment of the atrophic edentulous maxilla implant supported overdentures provide the opportunity to obtain proper lip support, speech and cleanliness. A superstructure with high rigidity is needed for an optimal distribution of occlusal loading, to ensure good prognosis for implant survival. A prosthodontic concept using a hybrid construction meets these criteria. It consists of a cast and precision-milled mesostructure fixed to the implants, over which that a removable cast prosthodontic device is placed to which the teeth are attached. It is essential to follow a strict treatment protocol to obtain a good and predictable result. PMID- 11924411 TI - [General discussion and summary]. PMID- 11924412 TI - [Viral hepatitis]. PMID- 11924413 TI - [Systemic antibiotics in the treatment of periodontitis]. PMID- 11924414 TI - [Rational use of antibiotics by the dentist can prevent microbial resistance]. AB - Worldwide resistance of microorganisms to antibiotics is becoming a problem of major concern. An antibiotic policy which tries to control the development of resistance by rational use is needed in all fields where antibiotics are prescribed. In dental practice, apart from endocarditisprophylaxis, prophylaxis is leukopenic patients and prophylaxis after dental implants, antibiotics can be used to support local therapy of an dentoalveolar abscess or refractory periodontitis. In the majority of these cases, a small spectrum penicillin is therapy of choice. PMID- 11924415 TI - [Masticatory muscles. Part III. Biomechanics of the masticatory muscles]. AB - The masticatory muscles are able to produce forces. These forces may cause movements of the lower jaw. Furthermore, they can be applied by the teeth for the generation of bite or chewing forces. During these kind of processes the temporomandibular joints will be loaded also. The interaction between forces and movements in the masticatory system is complex but obeys the relatively simple laws of mechanics. By application of these laws the development of joint loading, force patterns and movements during masticatory function and dysfunction can be understood. This is illustrated by a few examples of both statical and dynamical masticatory performance. PMID- 11924416 TI - [Aluminum oxide abutments for implants]. PMID- 11924417 TI - [Diseases of the pharynx: diphtheria]. PMID- 11924418 TI - [Dentistry, 50 years as an academic study. Introduction]. PMID- 11924419 TI - [Education and qualification of dentists in the Dutch legislation]. AB - Education and qualification of dentists are linked together in a certain way. The developments with respect to that are described, successively in the period before 1947 and afterwards, since the training of dentists became a university education. This improved education, leading to a broader qualification for the newly trained dentists. As a result of a new law handling of the obtained qualification by a dentist will be related to continuing education to be competent to practice dentistry according to the scientific state of affairs in dentistry. PMID- 11924420 TI - [Dentist and title. The importance of science for dentistry]. AB - Dentistry as a science is relatively new. In the Netherlands dental research has developed progressively during the last decades. This however, has only slightly influenced the training of dental students. In this article the interest of science to dentistry and dental training is explained from the point of view of dentistry as a developing science, from the point of view of the problem solving clinician and from the learning student. PMID- 11924421 TI - [The development of dental research in the Netherlands]. AB - From the acquisition of an academic status (ius promovendi) in 1947 till now, dental research in the Netherlands has reached maturity. An even increasing body of PhD-theses and publications have found their way towards the international scientific dental community. Since the early nineties most research groups have been brought together in the 'Interuniversitaire Onderzoekschool Tandheelkunde' (Netherlands Institute for Dental Sciences), a collaboration of the academic dental institutions in Amsterdam, Nijmegen and Utrecht. PMID- 11924422 TI - [Fifty years of dissertations by dentists]. AB - From 1947 the faculties of dentistry of the Dutch universities are entitled to grant Ph.D. grades for dental theses. In the fifty years that have elapsed since, about 500 dentists defended their thesis; the vast majority (6/7) during the second half of the period. This article describes the first thesis (dating from 1956-1973) of each of the five dental faculties that have existed in the Netherlands, followed by a short analysis of the entire group of 500 dissertations. In recent years dissertations are hardly ever written in Dutch, English taking over nearly entirely. Subjects in the field of social dentistry seem to loose foot. The increase in numbers of dental dissertations is stagnating (at a level of 20-30/year), just as is the case for medical theses (600 700/year). PMID- 11924423 TI - [Quality of the sterilization process in the dental practice]. AB - A survey was performed among 249 Dutch dentists on the use and functioning of sterilizers in their practice and on the quality control of the sterilization process. 99% of the dentists use a sterilizer. Only 1% uses only a thermodesinfector. 7.3% of the sterilizers tested did not function properly, based on a microbiological test. 76% of the sterilizers are not serviced on a regular basis. These sterilizers show more often failures in the sterilization process. It is concluded that although over 92% of the sterilizers in Dutch dental offices do function well, improvement on the quality control of the sterilization process is feasible and necessary. Besides an annual service, a regular control of the sterilization process itself, e.g. by using biological indicators, in combination with physical and/or chemical monitoring, is one of het methods to achieve this. PMID- 11924424 TI - [Masticatory muscles. Part IV. The masticatory muscles do not work homogeneously]. AB - The temporalis and masseter muscles have a complex architectural design with large attachment areas. As a consequence, each of these muscles is capable of producing a large number of mechanical effects. In addition, within each muscle the muscle fibres and sarcomeres undergo different excursions during jaw movements. This leads to intramuscular differences in the possible excursion range and the capacity to produce force. Depending on the desired motor task, the nervous system is able to vary both the magnitude and direction of muscle force by selective activation of specific muscle portions. The anterior temporalis and the superficial and deep masseter are capable of generating large forces, e.g. during biting and chewing. The posterior temporalis and the anterior and posterior deep masseter are suited to establish a precise adjustment of forces and movements during laterotrusion and protrusion/retrusion. PMID- 11924425 TI - [An unusual bump on the tongue]. PMID- 11924427 TI - [Masticatory muscles. Part V. Geometry of the masticatory muscles and cranial morphology]. AB - Studies focussed on the interactions between masticatory function and the variation in craniofacial height have received an important impulse by the availability of non-invasive imaging techniques like CT and MRI. These techniques allow for in vivo determination of the cross-sectional area and spatial orientation of the human jaw muscles. In recent MRI studies it has been established that the jaw muscles of long-face subjects are up to 30% smaller than those of normal individuals, while the position of the muscles was fairly comparable in both groups. The maximum bite force of long-face subjects is roughly half that of normals. The observed variation of the in vivo data of normal and long-face jaw muscle geometry does explain only half of the difference in their average maximum bite force. The jaw muscles of long-face and normal subjects are presumably different with respect to their force generating capacity per unit of cross-sectional area, which may be attributed to a different muscle fiber type composition. PMID- 11924426 TI - [The new fight against mutant streptococci]. AB - Mutans streptococci are an important factor in the aetiology of dental caries. New ways of combating these bacteria have become available with the development of molecular biological techniques. One of these strategies involves the genetic modification of mutans streptococci to develop less cariogenic mutants. Acid production and the production of intracellular polysaccharides have been eliminated. But it is not clear yet whether these less cariogenic mutant strains are able to replace the normal mutans streptococci in dental plaque. Immunization is another possibility to suppress mutans streptococci. Antibodies against proteins involved in adherence of mutans streptococci have been raised by active immunization. It is also possible to raise antibodies in cow serum and milk, in eggs of chickens and even in tobacco plants. After isolation, these antibodies are to be applied topically. In the future, these new strategies may prevent or reduce caries. However, further research is necessary to resolve the obstacles on the way to commercial application. PMID- 11924428 TI - [Photodynamic therapy of the oral mucosa]. AB - Photodynamic therapy (PDT) is a new therapeutic modality in head and neck oncology. PDT is based on the accumulation of a photosensitive dye in (pre)malignant tissue. When sensitizer containing tissue is exposed to light of a proper wavelength and dose, a photochemical reaction between the dye and the light will occur. The result may be tumornecrosis. Besides a tumor destructive effect, photosensitizers are also capable to show fluorescence, when stimulated by light of an appropriate wavelength. This fluorescence photo-detection (PD) can be used for early detection and localization of tumors or even premalignant epithelial changes, in tissues exposed to the delivered light. In this study several aspects of PDT and PD are described. PMID- 11924429 TI - [Changes in oral health in adults. Results of studies performed in 1983 and 1995]. AB - In 1995 a dental survey among adults aged 25-54 years was performed in 's Hertogenbosch (The Netherlands). Aim was to study trends in oral health after 1983, a year in which a similar study was performed in the same city. Caries data were obtained by clinical examination only. The percentage of edentulous persons in the sample decreased with about 50% between 1983 and 1995. In 1995 in all age categories of dentate persons, the mean number of FT was higher, and the mean numbers of DT and MT were lower than in 1983. In persons with a natural dentition under the age of 35 the mean number of DMFT decreased significantly. It was concluded that oral health in adults living in 's-Hertogenbosch, as measured by caries prevalence, treatment level of caries and percentage of edentulous persons, improved significantly between 1983 and 1995. It is supposed that the trends found in 's-Hertogenbosch are indicative of changes in oral health in The Netherlands. PMID- 11924430 TI - [Mental problems in the dental practice: a compulsive disorder]. AB - The aim of the present paper is to inform the reader about individuals who are suffering from severe mental disorders, such as obsessive-compulsive disorder (OCD). A case story enlightens the fact that anxiety disorders like OCD can have profound implications for the dental treatment plan. PMID- 11924431 TI - [The reproduction of the interocclusal relationship with the aid of a 'jig']. PMID- 11924432 TI - [Diseases of the pharynx: whooping cough]. PMID- 11924433 TI - [Maxillofacial trauma. How to treat?]. AB - Fractures of the maxillofacial skeleton, often accompanied by dentoalveolar injuries, are no exception. Dentists may become primarily involved in the diagnosis of such a trauma and the treatment of dentoalveolar injuries. More often they will play an important role in the aftercare of such a patient (endodontics, prosthodontics). This paper describes the general principles of the treatment of such patients. The specific treatment of maxillofacial trauma will be described in the contributions of other authors in this special issue. PMID- 11924434 TI - [Epidemiology of facial injuries]. AB - Epidemiological studies of maxillofacial trauma are numerous in the literature, but as a general rule their results are applicable to the investigated cohort only. Nevertheless, major differences in pattern of maxillofacial fractures have been reported between these studies. The longitudinal Nijmegen study (period 1960 1987) describes changes in the pattern of maxillofacial fractures for the Dutch situation. Traffic accidents still are the major cause of maxillofacial trauma, but the contribution of violent crimes and sports is rising. In addition, alcohol abuse as a factor in maxillofacial trauma has increased from 3% to 12% between 1960 and 1987. The current Dutch and international registration and classification systems need consent to allow for comparison of the results. Such systems should include ranking according to etiology, means of transport and age. Insight in the pattern of accidents, including maxillofacial trauma, will result in better ways of prevention of such accidents. PMID- 11924435 TI - [Concussion and contusion of the brain]. AB - A summary of the presentation, diagnosis and therapy of head injury is given. The article is focussed on mild head injury, the most frequent type of head injury. The difference between concussion and contusion is explained. Symptoms and abnormalities found at neurological examination are discussed. The Glasgow Coma Scale is described as a tool to diagnose the severity of head injury. The value of additional radiological investigation is discussed. The therapy which depends on the severity of head injury and concomitant complications, is given. Information about eventual persisting complaints and deficits is important. After care depends on the severity of the injury and the persistence of neurological signs and symptoms. Outcome measurement is done by the Glasgow Outcome Scale. It is emphasized that patients may exhibit persistent complaints although they return to work or school. PMID- 11924436 TI - [Soft tissue injuries of the face]. AB - Traumatic lesions of the soft tissues of the face may have an important social impact in affecting vital functions of the face as well as esthetics. Despite of some tolerance in adapting to tissue loss, major tissue of the face needs to be reconstructed at all levels. Primary treatment is most successful if reconstructions are performed atraumatically, using well vascularized tissues and respecting the esthetic units of the face. Functional structures such as the eye lids, the lips, the facial nerve and the parotid duct should be repaired as a primary procedure. Scar maturation is a prerequisite before performing secondary corrections. PMID- 11924438 TI - [Mandibular fracture]. AB - This article describes the present status of diagnostic and therapeutic methods to treat mandibular fractures. In view of the needs of a functional rehabilitation as complete as possible, considerations about treatment procedures are presented. Fracture treatment of the mandible in this respect means anatomical reduction and stabilisation of the fracture fragments in order to achieve complete restoration of the bony continuity, preferable without intermaxillary fixation, and restoration of speech and chewing functions enhanced by normal dental hygiene. The role and development of fixation techniques are described and discussed. Finally the role of the general practitioner at the time of diagnostics and initial treatment as well as regarding postoperative treatment and interpretation of treatment modalities is discussed. PMID- 11924437 TI - [Fracture healing of the face. General principals]. AB - To obtain good and uncomplicated primary fracture healing, anatomical repositioning, direct bone apposition and stable fixation are mandatory. The course of primary fracture healing starts with acute inflammation (cellular and vascular), stabilization (formation of new bone), and remodeling. Besides good fixation, local factors such as age and type of fracture will influence the healing. If good reposition, apposition or adequate stabilization of the bony fragments are not achieved, the bone healing will be secondary. In secondary healing fracture repair and stabilization occurs through the formation of callus. PMID- 11924440 TI - [Zygomatic fractures]. AB - The zygomatic bone is important for the midfacial contour and for protection of the orbital contents. Zygomatic fractures frequently occur. Because they are hidden behind a simple black eye in many instances, there is a potential risk of missing the diagnosis. Fresh zygomatic fractures are relatively easy to treat, but late secondary corrections are difficult and often have dissatisfying results. The most important clinical signs of a zygomatic fracture are: flattening of the zygomatic contour, edema, haematoma, unilateral nose-bleeding, disturbed sensibility, diplopia, restricted mandibular movements, contour defects. In this paper the clinical signs of a zygomatic fracture are emphasized to enable the general practitioner to examine the patient with a simple black eye appropriately and be able to recognize a zygomatic fracture. PMID- 11924439 TI - [Fractures of the condylar process]. AB - Fractures of the mandibular condyle are frequently seen. There is no consensus on the most appropriate treatment modality. Closed reduction with help of arch bars and guiding elastics is in most cases sufficient. Open reduction and fixation with a plate osteosynthesis or a specially designed lag screw is occasionally performed. Intracapsular fractures of the condylar head are only operated in case of severely reduced mobility, or ankylosis. Fractures of the mandibular condyle normally heal uneventfully. Complications mostly occur in bilateral condylar neck fractures, or intracapsular fractures. PMID- 11924441 TI - [Fractures of the midface]. AB - Midfacial fractures have a wide variety of appearance. The original classification by Le Fort is still in use. Nasal-orbital-ethmoidal (NOE) fractures deserve special attention. Next to clinical evaluation detailed computer tomography is necessary. Extended open reduction and internal fixation with various mini- and microplate osteosyntheses as well as immediate reconstruction with calvarian bone are essential components of surgical treatment aiming for adequate function and esthetics. Reduction of increased inner intercanthal distances is a substantial part of the treatment of NOE fractures. Surgical repair within 48-72 hours benefits the final result and limits the number and extent of secondary corrections. PMID- 11924442 TI - [Secondary posttraumatic correction of the face]. AB - Any part of the face may present residual deviations in form and/or function after trauma, even if primary treatment has been executed optimally. Secondary corrections may be indicated as a consequence. The techniques are usually the same as for primary treatment. However since secondary corrections quite often are a consequence of complicated primary problems, it does not surprise, that frequently complex deformities present for secondary surgery. Depending on the nature of the lesion percentages of success between 60 and almost a 100% may be expected. This indicates that additional treatment after primary care may be a way out for quite a number of residual deformities. PMID- 11924443 TI - [Dietetic care for patients after maxillofacial trauma]. AB - Patients suffering from maxillofacial trauma and subsequent intermaxillary fixation are temporary at risk of severe malnutrition and substantial weight loss. Eventually, these consequences may lead to impaired fracture healing and wound healing, and thus to deteriorated overall functional recovery. Adequate evaluation of several specific risk factors warrants the opportunity to stratify individual patients to be at either high or low risk for nutrition related health problems. Customary dietary assessment and counselling can be provided when necessary. PMID- 11924444 TI - [Bacterial meningitis]. PMID- 11924445 TI - [The cerebellum: from motor coordination to cognitive function]. AB - Clinical data in man, as well as experimental results in animals, classically involve the cerebellum in the coordination of ballistic movements and in their accompanying postural adjustment. The cerebellum intervenes in the coding of the order and duration of contraction of the different protagonist muscular groups contributing to the same movement. In normal life, this is an automatic, non conscious procedure. Recent studies seem to indicate that the human neocerebellum (lateral hemispheres and dentate nuclei) plays a role in the regulation of some neocortical cognitive functions. This new functional aspect of cerebellar activity has been inferred from the results obtained by three quite different domains: neuroanatomical data showing the existence of, sometimes reciprocal, pathways between the neocerebellum and associative and limbic areas in primates, neuropsychological data assessing the presence, in some cerebellar patients, of purely cognitive impairments, and data from functional imagery pointing out cerebellar activation in healthy subjects during non motor tasks. II would ensue that, thanks to new cortical targets. The cerebellum could regulate sensorial, procedural, linguistic and emotional activities, so that a cerebellar lesion could be followed by a cognitive and affective syndrome, depending on the importance and on the location of the lesion. PMID- 11924446 TI - [Treatment of newly diagnosed epileptic crises. A French experience]. AB - CAROLE is a prospective survey of 1942 children and adults presenting at least one unprovoked epileptic seizure first diagnosed between May 1 1995 and June 30 1996 by 243 French neurologists and neuropediatricians. In half these cases, the patient had already experienced more than one seizure at the time of diagnosis. Patients presenting only one seizure but either a case history or significant EEG or neuroimaging abnormalities suggestive of a high risk of recurrence were classified as cases of epilepsy. Drug treatment was prescribed to 73 p. 100 of the cohort. Treatment was far more frequent in cases where seizure had recurred (90 p. 100) than in cases of first seizure (54 p. 100) (p < 0.0001). In the latter category, the older the patients, the more likely they were to be treated (69 p. 100 of patients > or = 60 years old versus 49 p. 100 of those under 24 years of age, p = 0.001). Those with multiple onset were more often treated than those with single onset (48 p. 100 of cases of single seizure versus 80 p. 100 of clustered seizures and 90 p. 100 of status epilepticus, p < 0.0001), as were cases of complex partial seizure (68 p. 100 versus just under 50 p. 100 of cases of generalized seizure, p = 0.0001), those with epileptiform EEG abnormalities (67 p. 100 of cases of cryptogenic partial epilepsy and 63 p. 100 of idiopathic generalized epilepsy versus 42 p. 100 of isolated seizure, p < 0.0001) and those with cerebral lesions visible by neuroimagery (89 p. 100 of cases of symptomatic partial epilepsy). The decision not to treat was generally based on clinical signs of non-gravity. Understandably, cases of idiopathic partial epilepsy were less treated than other syndromes, 22 p. 100 in the group of first seizure patients, 67 p. 100 in the group where seizure had recurred before diagnosis. A single-drug therapy was recommended to 1389 of the 1411 patients treated, in 63 p. 100 of cases sodium valproate, in 30 p. 100 carbamazepine. These data reveal not only the attitude to treatment of the specialists concerned, but also the desires or refusals of patients or patients' parents. The present study was limited to this question of treatment or no treatment. A further study will be necessary to investigate to what extent patients followed the treatment prescribed and how these cases evolved. PMID- 11924447 TI - [Procedural learning and Parkinson disease: implication of striato-frontal loops]. AB - OBJECTIVE: To investigate procedural learning in non demented patients with idiopathic Parkinson's disease (PD). BACKGROUND: Experimental evidence implicate the basal ganglia in procedural learning. Selective impairment has more recently been described in patients with frontal lesions. METHODS: The performance of 20 demented patients and 15 matched normal controls was studied in the serial reaction time task (SRTT). Performance on procedural task was further compared with that of 9 normal controls and with patients' performance on tests assessing explicit memory, executive functions and global efficiency. RESULTS: The group of patients with PD showed impaired procedural learning. The difference of response time between the repeated and the non-repeated blocks was smaller in PD when compared to controls. Subsequent analyses separated PD patients into two subgroups according to their performance on SRTT, measured by the rebound effect. PD patients whose learning was normal differed from PD patients whose learning was impaired on performance in tests sensitive to frontal lobe dysfunction. CONCLUSION: These results confirm the implication of the striatum in procedural learning and suggest that performance on cognitive procedural learning depends on the striato-frontal circuits. PMID- 11924448 TI - [Idiopathic SUNCT (short lasting unilateral neuralgiform headache attacks with conjunctival injection, tearing, sweating and rhinorrhea) syndrome: 2 new cases]. AB - The syndrome of short-lasting unilateral, neuralgiform attacks of pain in the peri orbital area associated with conjunctival injection and tearing (SUNCT) is a rare disorder affecting mainly males. We report two French patients (1 male and 1 female) with SUNCT syndrome, 27 and 28 years of age respectively. Both patients had short (30 sec), frequent (30-100/day) excruciating pain located at the peri orbital area, associated with conjunctival injection, tearing, rhinorrhea, ptosis and others vasomotor symptoms. Clinical examination and imaging were normal. Most drugs used in the treatment of migraine, cluster headache, trigeminal neuralgia, and other short-lasting headaches were not successful. PMID- 11924449 TI - [Identification of factors influencing hospital admission delay after ischemic cerebrovascular stroke. Study of a rural population]. AB - We studied the time of arrival of 235 consecutive patients admitted to the emergency department of a University Hospital located in a rural area after the first symptoms of ischemic stroke or TIA. Among the following factors, we determined those that might be involved in delayed admission: place of symptom onset, time and place of onset of the first symptoms, contact with a general practitioner before admission time, mode of transportation, clinical score, impairment of consciousness, presence of seizures, heart complaints or headache, age and past medical history of cerebrovascular, cardiovascular and hypertension diseases. Half of the patients arrived within 4 h 10 of symptom discovery and 55 p. cent arrived within 6 hours. The percentage of patients arriving within 3 h (p = 0.001) and 6 h (p = 0.001) was higher for those who had a stroke during the day (8 a.m.-8 p.m.) than during the evening and night. The other characteristics associated with a shorter delay included a low neurological score on the Mathew's Stroke Scale (p < 0.001 at 3 h and p = 0.001 at 6 h) and younger age (p = 0.015 at 3 h). Presence of headache delayed admission (p = 0.010). Forty-five percent of patients arrive at the hospital 6 hours after the discovery of symptoms, too late to receive optimal stroke therapy. Widespread public education on stroke is necessary to reduce the delay of admission, particularly for old patients and in case of mild to moderate deficits. PMID- 11924450 TI - [Cerebral glioblastomas and systemic metastases]. AB - Glioblastomas are highly malignant but locally invasive tumors. In only rare instances do extra-neural metastasis occur. We report herein 2 cases of such dissemination occurring 9 and 15 month after the initial diagnosis and treatment of supratentorial gliomas. Metastasis involved the lung, the kidney and the liver in one case and the bones in the other case. Both patients died within 3 month after diagnosis of the systemic metastasis despite chemotherapy. PMID- 11924451 TI - [Susac syndrome in a man]. AB - The association of an acute encephalopathy, neurosensory hearing loss and retinal branch artery occlusions in a 35 year old man as well as the absence of systemic disease suggest the diagnosis of Susac's syndrome. This is the eighth male case described, the syndrome being more frequently reported in females. PMID- 11924452 TI - [Signet-ring cell carcinomatous meningitis complicating esophageal neoplasia]. AB - Leptomeningeal invasion is an uncommon complication of systemic solid cancers. We describe a patient with such an infiltration caused by a signet-ring cells esophageal adenocarcinoma. The diagnosis required repeated cytologic examinations of cerebrospinal fluid while clinical signs and biological data were not conclusive, mimicking acute infectious meningoencephalitis. Magnetic resonance imaging was highly contributive in establishing the diagnosis. Nevertheless this is a very rare complication of esophageal cancers as this metastatic location occurred in advanced stages of the disease and as we know the poor prognosis of those cancers. PMID- 11924453 TI - [Secondary subdural hematoma in dural metastasis of prostatic adenocarcinoma]. AB - Non traumatic subdural hematoma secondary to dural metastases is a rare event but is well documented. We report a case of a 73-year-old man who presented a subdural hematoma secondary to dural metastases from prostatic adenocarcinoma. The diagnostic was made during the operation. The bone and the dura-mater were infiltrated by tumor cells. Mechanisms of transdural invasion by metastatic adenocarcinoma and production of hematoma are discussed and the relevant literature is reviewed. PMID- 11924454 TI - [Carpal tunnel syndrome with an unusual cause: a malignant nerve sheath tumor of the median nerve]. AB - We report on the follow-up of a patient who developed symptoms suggestive of carpal tunnel syndrome. Symptoms were however atypical with involvement of the nondominant hand and with selective, fascicular, electroneurographic changes. During the surgical decompression of the median nerve at the wrist a tumor was found, corresponding to an isolated malignant peripheral nerve sheath tumor (MPNST) of mild type. A course of local radiation therapy was completed, with no sign of recurrence, and a normalization of the serum level of neurone specific enolase. PMID- 11924455 TI - [Epilepsy surgery followed by schizophrenia]. PMID- 11924456 TI - [Paraneoplastic peripheral neuropathies]. PMID- 11924457 TI - [Lewis and Sumner syndrome]. PMID- 11924458 TI - [Pain in peripheral neuropathies: physiopathology, clinical evaluation and medical management]. PMID- 11924459 TI - [Neurophysiologic exploration of the brachial and lumbosacral plexus]. PMID- 11924460 TI - [Simultaneous bilateral paralysis of cranial nerves V and VI unmasking neurosarcoidosis: symmetrical involvement of Meckel's cave]. PMID- 11924462 TI - Rapid testing stalled. PMID- 11924461 TI - [Sensory disorders in multiple sclerosis (before optic neuropathy)]. PMID- 11924463 TI - You don't own me. PMID- 11924464 TI - Rapid new world. PMID- 11924465 TI - Patent primer. PMID- 11924466 TI - Independent review. PMID- 11924467 TI - HIV & breast feeding: what's a mother to do? PMID- 11924468 TI - [Danish physicians and smoking habits among the Danes. What to do?]. PMID- 11924469 TI - [Should blood pressure be regulated during the acute phase of apoplexy?]. PMID- 11924470 TI - [Treatment of nicotine addiction. Drug therapy for smoking cessation]. AB - Most Danish smokers are addicted to nicotine. Treatment with nicotine replacement therapy or bupropion doubles the one-year success rate after cessation of smoking. Pregnant smokers who are unable to stop smoking without medical treatment can safely be offered nicotine replacement therapy. The health risk of smoking is particularly high in patients with cardiovascular disease. Reluctance to use nicotine products for patients with heart disease is not supported by scientific evidence, and either bupropion or nicotine products can safely be administered to them. PMID- 11924471 TI - [Retirement plan for a 70-year-old. Intravenous urography disembarks from uroradiology]. AB - Over the last 70 years, intravenous urography (IVU) has played a major role in the work-up of diseases in the kidneys and the upper urinary tract. However, modern cross-sectional modalities have shown that IVU does not fulfill current requirements. Patients with renal colic are better examined by non-enhanced CT scanning, as it finds more stones (60 versus 100%). Furthermore, a disease outside the upper urinary tract is found in about 15% of patients with renal colic, and in 3% another kidney or upper urinary tract disease is found. For patients with haematuria, CT urography is a better alternative, as it combines the advantages of CT with those of IVU. IVU misses about 15% of renal tumours above 3 cm in diameter and almost all below 3 cm. MR urography is the newest way of imaging the upper urinary tract; its exact role is still undetermined, but it is recommended for children, young persons, and pregnant women. After 70 years of good performance, it is time for IVU to retire. PMID- 11924472 TI - [Strategy for peroperative fluid therapy--balancing the central blood volume]. AB - The strategy for preoperative volume treatment is not clear, partly because of the difficulty in monitoring the central blood volume (CBV). The cardiovascular responses to hypovolaemic shock are reviewed with emphasis on the often very low heart rate, which develops when the CBV is reduced by about 30%. A proposal has been put forward that normovolaemia is the CBV that does not limit cardiac output. The possibility of monitoring CBV by electrical impedance is also reviewed. It is concluded that the deviations reflect the CBV accurately in both experimental and clinical studies. A strategy that maintains the CBV ensures cerebral oxygenation even during extensive haemorrhage. PMID- 11924473 TI - [Ambulance transportation and prehospital treatment in connection with admission for suspected acute myocardial infarction]. AB - INTRODUCTION: The aim was to describe ambulance transportation and pre-hospital treatment in connection with admission for suspected acute myocardial infarction. MATERIAL AND METHODS: For all patients with suspected acute coronary syndrome who were urgently admitted to the Cardiological Department, Odense University Hospital between 3 August 1998 and 6 December 1998, information about ambulance transportation and pre-hospital treatment was collected through interviews with the patients and study of ambulance records, admission notes, and hospital medical records. In addition, details of the regarding response times were obtained from Falck's emergency service and from nurses' papers. RESULTS: Altogether 279 patients (83%) were transported by ambulance. Half the ambulances arrived at the hospital after 34 minutes (range 11-140 minutes), but every third ambulance took more than 40 minutes to reach the hospital. The pre-hospital treatment of all the patients was: oxygen 69%, nitroglycerin sublingually 46%, nitrous oxide 2%, defibrillation 1.4%, acetylsalicylic acid 9%, morphine injection 8%, and ECG monitoring 57%. CONCLUSION: The study showed that there were quality problems, as every third ambulance took more than 40 minutes to reach the hospital. It also showed that acetylsalicylic acid and morphine were used only to a limited extent in a pre-hospital situation. PMID- 11924474 TI - [The relation between life style, socioeconomic factors, social networks and suboptimal self-assessed health]. AB - INTRODUCTION: The aim of the present study was to assess the relation between these life-style factors and suboptimal self-reported health. METHODS: A cross sectional study was carried out on a random sample of 12,040 men and women from Copenhagen, who had answered a questionnaire regarding smoking, alcohol intake, physical activity, socio-economic status, social network, and self-perceived health. RESULTS: Smoking, high alcohol intake, and physical inactivity are strongly associated with a suboptimal self-reported health. DISCUSSION: The assumption of a correlation between a high quality of life and a shorter life cannot be confirmed by this study. PMID- 11924475 TI - [Mortality associated with physical activity in leisure time, at work, in sports and cycling to work]. AB - INTRODUCTION: Previous studies have shown that high levels of physical activity are associated with lower mortality in middle-aged men. Few studies have investigated this association in women and the independent effects of cycling to work and participation in sports. MATERIAL AND METHODS: The participants were 13,445 women and 17,441 men aged 20 to 93 years. Self-reported physical activity included general questions about leisure time physical activity and physical activity at work, sports participation, and cycling to work. Adjustment was made for blood pressure, total cholesterol, triglyceride, body mass index, smoking, and educational level. RESULTS: During 433,000 person-years of observation, 2738 women and 4672 men died. Physical inactivity during leisure time predicted mortality in both men and women in all age groups. In women and men, the most physically active in leisure time experienced only half the mortality of the sedentary. Even in the moderately and highly active persons, sports participants experienced only half the mortality of non-participants. Physical activity at work predicted mortality in women only. The men and women who rode a bicycle to work had a 39% lower risk of mortality after multivariate adjustment including leisure time physical activity. DISCUSSION: Leisure time physical activity was inversely associated with all-cause mortality in both men and women in all age groups. Benefit was found from moderate leisure time physical activity with further benefit from sports activity and bicycling as transportation. Benefit from physical activity at work was found in women only. PMID- 11924476 TI - [Qualitative assessment of pain relief and functional improvement after coronary bypass surgery. A questionnaire survey among 527 patients]. AB - INTRODUCTION: We wanted to assess postoperative pain relief and functional improvement in all CABG patients (546) from Gentofte County Hospital (GCH) in 1997, so as to elicit better information on what to expect after CABG. MATERIAL AND METHODS: A descriptive cross-sectional survey, was carried out in 1999. All surviving patients (527) were sent a questionnaire with 17 questions in five categories: The course of the operation, pain level, medicine consumption postoperatively, working situation, and educational level. The inclusion criterion was CABG in GCH in 1997. RESULTS: One hundred and five women and 441 men (mean age 74 and 67 years) were entered. Response rate: 98%. Mortality rate: 0.9% in 30 days and 3.5% in two years. Sixty-four percent were satisfied with the course of the operation. In two years 33% were rehospitalised and 6% had a new CABG/PTCA. Other frequent diagnoses for readmission were AMI, arrhythmia, and angina. Considerable wound discomfort was reported by 21% (chest) and 13% (leg/arm). Marked pain relief and functional improvement was achieved postoperatively (95%). Postoperative angina was not associated with limitation in daily activities. Postoperative medicine consumption: 52% antihypertensives and 20% anti-angina medicine. The return-to-work rate, for those working before the operation was 75% in about two months, with a slight preponderance in those employed in the private sector. DISCUSSION: The study shows a high survival rate in two years after CABG. Considerable pain relief and functional improvement was achieved, and only a very small group derived no benefit from the operation. A large amount of patients had wound discomfort, which leaves an unsolved problem. PMID- 11924477 TI - [Allergic contact eczema because of diethylthiourea in neoprene rubber]. AB - Three cases are reported of acute allergic contact dermatitis caused by diethylthiourea in the neoprene rubber used in a diving suit and orthopedic braces. Patch testing should be done with the specific thiourea compounds, because testing with material from the suspected products may be negative. Cross reactivity between thiourea compounds is uncommon. PMID- 11924478 TI - [Gabapentin treatment of glossopharyngeal neuralgia with cardiac syncope]. AB - A 61-year-old man with glossopharyngeal neuralgia (GPN) associated with cardiac syncope was successfully treated with gabapentin. The treatment of GPN with the antiepileptics is discussed with focus on gabapentin, because of its few side effects in comparison with other anticonvulsants. PMID- 11924479 TI - [Blood pressure determination]. PMID- 11924480 TI - [Quality assurance of treatment of ischemic syndrome at the Fredericia hospital]. PMID- 11924481 TI - [Weight control and physical activity--significance for the development of cancer]. PMID- 11924482 TI - [Away with the lagging behind when it comes to the average length of life]. PMID- 11924483 TI - [Back pain]. PMID- 11924484 TI - [Hepatitis C treatment without liver biopsy]. PMID- 11924486 TI - [Driver's licence--again]. PMID- 11924485 TI - [Placebo and the evidence burden]. PMID- 11924487 TI - [Management of stomas]. PMID- 11924488 TI - [Written declaration]. PMID- 11924490 TI - Theory and practice of parallel direct optimization. AB - Our ability to collect and distribute genomic and other biological data is growing at a staggering rate (Pagel, 1999). However, the synthesis of these data into knowledge of evolution is incomplete. Phylogenetic systematics provides a unifying intellectual approach to understanding evolution but presents formidable computational challenges. A fundamental goal of systematics, the generation of evolutionary trees, is typically approached as two distinct NP-complete problems: multiple sequence alignment and phylogenetic tree search. The number of cells in a multiple alignment matrix are exponentially related to sequence length. In addition, the number of evolutionary trees expands combinatorially with respect to the number of organisms or sequences to be examined. Biologically interesting datasets are currently comprised of hundreds of taxa and thousands of nucleotides and morphological characters. This standard will continue to grow with the advent of highly automated sequencing and development of character databases. Three areas of innovation are changing how evolutionary computation can be addressed: (1) novel concepts for determination of sequence homology, (2) heuristics and shortcuts in tree-search algorithms, and (3) parallel computing. In this paper and the online software documentation we describe the basic usage of parallel direct optimization as implemented in the software POY (ftp://ftp.amnh.org/pub/molecular/poy). PMID- 11924491 TI - Some statistical aspects of the maximum parsimony method. AB - The last three decades have seen considerable debate concerning the relative merits and problems associated with two competing approaches to phylogeny- approaches based on the parsimony principle versus maximum likelihood methodology. Although the two approaches may seem quite opposed, there are in fact some close relationships between them. For example, we describe a recent result that shows how maximum parsimony can be regarded as a type of maximum likelihood estimator when there is no common mechanism between sites (such as might occur with morphological data and certain forms of molecular data). Distinguishing between this and other implementations of maximum likelihood helps clarify some of the dispute that has surrounded the two methodologies. We also provide a brief overview of some mathematical and statistical properties of the maximum parsimony criterion. PMID- 11924489 TI - DNA multiple sequence alignments. AB - In this chapter we examine the procedure of multiple sequence alignment. We first examine the heuristic procedures commonly used in multiple sequence alignment. Next we examine sources of ambiguity involved in the alignment procedure. We suggest that several alignment parameters be employed to examine alignment sensitivity. We end by presenting an experiment with humans showing the ambiguity involved in manual alignment. PMID- 11924492 TI - 'Pluralism' and the aims of phylogenetic research. AB - In science, and particularly in the field of phylogenetic systematics, investigators may choose among different methods to analyze their data. These methods include neighbor-joining (or other genetic distance approaches), maximum likelihood, and cladistic parsimony, among others. These distinct methods of analysis differ considerably in how they process information from the observed data. However, many published molecular analyses utilize trees generated under more than one of these methods, which we will call a 'pluralistic' approach. Here, we explore the statistical, philosophical and operational aspects of the pluralistic approach. We suggest that the pluralistic approach is misguided from all three perspectives and we propose an alternative, logically consistent, strategy as an aim of phylogenetic research. PMID- 11924493 TI - Molecular systematics and the origin of species: new syntheses or methodological introgressions? AB - The advent of molecular phylogenetics stimulated the need to reprise for many discussions surrounding species concepts. The interpretation of cladograms as accurate representations of phylogeny, when the characters upon which they are based exhibit a reticulate pattern, is inconsistent with the epistemological axiom of hierarchy we assign to the cladistic method (Brower, 2000c). Discrepancies in the interpretation of cladograms would appear to account for differences in the kinds of questions to which they are applied. The philosophical and empirical issues surrounding this subject are examined in this chapter. PMID- 11924494 TI - Is morphology still relevant? AB - The utility of morphological data in modern systematics has recently been challenged because strong selection pressures are thought to create widespread patterns of convergent evolution at this level. This concern has led to suggestions that morphological data should be excluded either from all analyses or at least from analyses where there is conflict with molecular data. These concerns, however, are generally unwarranted and excluding data is not a defensible strategy for dealing with problems that do exist. We emphasize the importance of empirical responses, such as collecting additional and diverse data and exploring taxa and data set interactions, rather than the implementation of a priori assumptions, to overcoming many of the concerns associated with combining morphological and molecular data. Numerous factors may create biases in both molecules and morphology. While these biases are prevalent enough to cause widespread incongruence, they highlight the importance of combining, rather than separating, data. Morphological data also offer distinct advantages over molecular data, such as the inclusion of fossil taxa, cost-effectiveness and presence of biases different from those in molecular data. PMID- 11924495 TI - Species to genera: phylogenetic inference in the Hawaiian Drosophilidae. AB - Systematic studies at any taxonomic level require careful planning, even before genes are sequenced or morphological characters scored. Molecular systematists working at the level between species and genera must select gene regions which are suitable for the divergence within the group being examined and decide how many and which ingroup and outgroup taxa to sample in the analysis. This chapter will use studies of the Hawaiian Drosophilidae to illustrate strategies useful in (1) selecting nucleotide sequences for divergences between the levels of species and genera, (2) designing ingroup and outgroup taxon sampling schemes, and (3) performing phylogenetic analysis on data sets with large numbers of taxa and characters. PMID- 11924496 TI - Development, homology and systematics. AB - The basic issue of recognizing and delimiting characters derived from different levels of biological organization, including molecular, cellular, morphological and behavioral levels, has been addressed previously in the cladistic literature. But when considering new sources of information (such as the proliferating Evo Devo data), it is critical to review traditional theoretical and methodological approaches to their interpretation. This is especially important because the conclusions of a phylogenetic analysis are dependent upon the initial recognition and definition of the characters, the basic units of comparison in phylogeny reconstruction. This chapter explores the role of recent Evo-Devo studies in systematics and attempts to place the Evo-Devo literature into a systematics context. PMID- 11924497 TI - Gene family phylogenetics: tracing protein evolution on trees. AB - How have proteins taken on the remarkable diversity of biochemical and physiological functions necessary to create and maintain complex organisms? The majority of proteins are organized hierarchically into families and superfamilies, reflecting an ancient and continuing process of gene duplication and divergence. The techniques of molecular phylogenetics, developed to recover the nested hierarchy of taxa from character information in their gene sequences, can also reconstruct the evolutionary relationships among genes and provide a conceptual foundation for comparative evolutionary analysis of proteins and their functions. In this review, I outline the application of phylogenetic approaches to issues in gene family studies, beginning with the inference of phylogeny and the assessment of the two types of homology by which genes in a family can be related: orthology (common descent from a cladogenetic event) and paralogy (common descent from a gene duplication event). I show how the phylogenetic approach makes possible novel kinds of comparative analysis, including detection of exon shuffling, reconstruction of the evolutionary diversification of gene families, tracing of evolutionary change in protein function at the amino acid level, and prediction of structure-function relationships. A marriage of the principles of phylogenetic systematics with the copious sequence data being generated by molecular biology and genomics promises unprecedented insights into the nature of biological organization and the historical processes that created it. PMID- 11924498 TI - Evolution of spider silk proteins: insight from phylogenetic analyses. AB - Spider silks have astounding mechanical properties. In fact, dragline silk has greater tensile strength than commonly used synthetic materials such as nylon filament and capture spiral silk is among the most elastic protein known. The recent cloning of spider silk genes has revealed that silk proteins are composed of tandem arrayed ensembles of a small number of amino-acid sequence motifs. These repetitive motifs form the structural modules within silk fibers, and ae critical for determining the mechanical attributes of the silk. In this chapter, I examine the evolution of these motifs in the 11 published spider silk gene sequences. Extensive rearrangements of the motifs have occurred among the orthologous and paralogous proteins. Phylogenetic analyses suggest that numerous length mutations and recombination events have taken place in orthologous genes from closely related species and even within sets of alleles from the same species. Such genetic events appear to be critical for the homogenization of amino acid repeats within the silk proteins. The characterization of additional silk genes will clarify the relationships among novel amino acid motifs, the homogenization of motifs within a protein, and the function of silk fibers. PMID- 11924499 TI - Comparative methods and evolution. AB - Comparative biology provides a framework for the study of evolution, by seeking answers to the question of why traits evolved. However, the difficulties of making inferences about the biological causes of trait variation and covariation resulted in the development of very different approaches to comparative analyses, which are outlined and contrasted here. The Homology approach differs from all other approaches as it attempts to explain historical uniques, whereas the various homoplasy approaches draw inferences from repeated origination of correlated traits. A distinction is made between those approaches deriving information from correlated transitions of characters on a cladogram (Homoplasy I) and those that simply extract correlated differences between pairs of phylogenetically independent taxa (Homoplasy II). The latter approach has been implemented in well-developed analytical procedures ("comparative methods") which, however, are principally non-historical and thus their inferences about evolution are indirect. In contrast, the study of correlation of character transitions seeks to explain trait variation as the outcome of evolution, but is hampered by the difficulty of reconstructing ancestral character states, a prerequisite for the analysis of correlated transitions. Model-based techniques for ancestral state reconstruction are being developed, but the lack of data contained in single characters limits the biological reality of parameter intensive models. PMID- 11924500 TI - The use of physiological data to corroborate cospeciation events in symbiosis. AB - The symbiotic association between sepiolid squids (Family Sepiolidae) and luminous bacteria (Genus Vibrio) provides an unusually tractable model to study the evolution and speciation of mutualistic partnerships. Both host and symbiont can be cultured separately, providing a new avenue to test phylogenetic congruence through molecular and physiological techniques. Combining both molecular and morphological data as well as measuring the degree of infectivity between closely related pairs can help decipher not only patterns of co speciation between these tightly linked associations, but can also shed new light on the evolution of specificity and recognition among animal-bacterial associations. PMID- 11924501 TI - Reexamining microbial evolution through the lens of horizontal transfer. AB - Our ability to understand the evolution of microbial organisms revolves around a central and increasingly unsettled question: what is the nature of the mode of inheritance? The extent to which genetic information is passed vertically from parent to daughter or horizontally between distant relatives must guide reconstructions and inferences of evolutionary history, and has direct bearing on any ideas about the mechanisms of selection and diversification. Recent evidence suggests that we may have previously underestimated the contribution of horizontal gene transfer, and the dynamics and extent of this process are only beginning to be understood. The recent flood of complete genome sequences of microorganisms has already presented us with a vast array of data from which to test our hypotheses about the evolution of the entire tree of life, but what remains unclear is how we can make sense of this unwieldy data set. Analyses of this newly available data set should include explicit examinations of the contributions of both types of inheritance. PMID- 11924502 TI - Higher-level systematic analysis of birds: current problems and possible solutions. AB - Avian systematics has a rich history, as evolutionary biologists have long been interested in this conspicuous and diverse group of vertebrates. Many prominent scientists, and evolutionary biologists in particular, have focused their efforts on birds. Perhaps no other group of vertebrates is so well studied. Yet, despite the attention paid to this group, much about the history of the class Aves remains controversial, both with respect to the origin of birds and the history since that origin. This puts avian systematists in a unique position, with so much information available and so many unanswered questions to pursue. The fact that avian ordinal relationships are still the center of much controversy speaks to the difficulty of the problem. While many prominent morphologists have worked on avian relationships, relatively few morphological studies have identified characters with informative variation for interordinal relationships. Molecular data offer the hope for phylogenetic information not present (or not discovered) in avian anatomy. Since the first study of avian proteins for the purposes of systematics (Sibley, 1960), several prominent molecular systematists have devoted tremendous time and resources to solving the problems of avian relationships using molecular characters (see Barrowclough, 1992 and Sheldon and Bledsoe, 1993 and references therein). So far, their efforts have not produced adequate resolution, at least not in the minds of most practicing systematists. Here, we first outline what we think we do know about higher order avian systematics and discuss some specific cases of molecular data applied to this question. Next, we consider some of the problems which may be blocking a clearer understanding of avian relationships. We then go on to offer some new directions for systematists working on this difficult group. PMID- 11924503 TI - Relative quality of different systematic datasets for cetartiodactyl mammals: assessments within a combined analysis framework. AB - High congruence, support, stability, resolution and decisiveness are seen as positive attributes by many systematists. Within a cladistic context, the consistency index, the retention index, branch support, data decisiveness, the number of nodes resolved in a strict consensus tree and the incongruence length difference are direct measures of these qualities. Phylogenetic analyses of 29 datasets for cetartiodactyl mammals show that for a particular character partition, these indices can vary radically in separate versus combined analysis of datasets. The quality of any single dataset is of little importance in comparison to a thorough sampling of the available character space. PMID- 11924504 TI - Cladistics, populations and species in geographical space: the case of Heliconius butterflies. AB - The paradox of the species in evolutionary thought has promoted much debate and numerous incompatible definitions and concepts. This chapter argues that although the phylogenetic species concept (the author's version of it, at least) is no more accurate a description of "speciesness" than any other species concept (indeed, the notion of accuracy is irrelevant, as will be seen), it links species definition to species diagnosis via explicit criteria, which renders phylogenetic species more amenable to empirical testing than species defined by other concepts. The practical implications of cladistic species concepts for determining the boundaries between geographically differentiated sister taxa are explored using the example of Heliconius, based on my work and the recent studies of Mallet and others. The problem of circumscription is also addressed, with particular reference to the concepts of subspecies and geographical races. PMID- 11924505 TI - Phylogeny of the holometabolous insect orders based on 18S ribosomal DNA: when bad things happen to good data. AB - The purpose of this chapter is two-fold. First, all available 18S rDNA sequences for the Holometabola to reappraise their phylogenetic relationships will be compiled. Second, these data and analyses will be used to highlight general problems in using molecular data to infer higher-level phylogeny. PMID- 11924506 TI - Relationships among metazoan phyla as inferred from 18S rRNA sequence data: a methodological approach. AB - The relationships among the phyla of Metazoa have been investigated by several authors. Different genes have been applied to this problem, but only the ribosomal gene 18S rRNA has been investigated for enough phyla so as to attempt an answer to the question of how the current living forms are related to each other (only one phylum, the Loricifera, is missing). In this chapter, I propose an alternative way to analyze the data obtained from ribosomal genes, or other non-coding genes that show sequence length variation. PMID- 11924507 TI - [What can we do with cyclodextrins?]. PMID- 11924508 TI - [Effect of cyclodextrins on fungal degradation of fluorene]. AB - The purpose of this study was to improve the bioavailability of fluorene (PAH) by the use of complexing agents, cyclodextrins. The biodegradation tests were performed in liquid medium batches; fluorene was quantified by HPLC. Experimental results showed the enhancement of fluorene degradation by Penicillium italicum and Phanerochaete chrysosporium in the presence of branched cyclodextrins. PMID- 11924509 TI - [Reduction of the volatility of volatile organic compounds with cyclodextrins: application to their capture]. AB - The interactions existing between beta-cyclodextrin, hydroxypropyl-beta cyclodextrin and six volatile organic compounds (VOC) were studied and used for capture of such pollutants. The reduction of volatility for these VOC appeared to be equivalent for the two kinds of cyclodextrins at equal concentrations, while a higher solubility conferred higher efficiency to hydroxypropyl-beta-cyclodextrin. The use of such cyclodextrins in traps allowed a two-fold increase in VOC capture compared with genuine water. Application of this process to an industrial site showed its efficacity qualitatively while several optimization procedure are needed to achieve quantitative efficacy. PMID- 11924510 TI - [Use of cyclodextrins in the formulation of polyalkylcyanoacrylate nanoparticles charged with different active ingredients]. AB - Cyclodextrins were used to improve the loading capacity of biodegradable pol(yisobutyl cyanoacrylat)e nanoparticles, which were obtained by anionic polymerization in aqueous medium. We investigated the feasibility of blank nanoparticles in the presence of a series of cyclodextrins (5 mg/ml) and poloxamer 188 (1%). The smaller particles (87 +/- 3 to 103 +/- 6 nm) were obtained in the presence of hydroxypropyl beta- or gamma-cyclodextrin. The nanoparticle loading capacity investigated in the presence of hydroxypropyl beta cyclodextrin, in the previous conditions, on a series of steroids revealed an increase varying from 5.5 times (megestrol acetate) to 130 times (prednisolone). Differential scanning calorimetry study of the active ingredient (progesterone) in the nanoparticles, revealed an amorphous or molecular state. The in vitro release of the active ingredient occurred very rapidly but reached a plateau depending on the nanoparticle size and the dissolution medium nature. All the active ingredient was released in the presence of esterases. The addition of a preformed hydroxypropyl beta-cyclodextrin/saquinavir inclusion compound to the preparation medium of poly(isobutyl [or] isohexyl cyanoacrylate) nanoparticles, resulted in a 20-fold increase in the encapsulation yield. Presently, poly(isobutyl cyanoacrylate) hydroxypropyl beta-cyclodextrin combined nanoparticles loaded with doxorubicin are in phase II clinical trials. PMID- 11924511 TI - Novel approach to the study of the chiral discrimination. Mechanism in a series of imidazole derivatives using HPLC. AB - In high performance liquid chromatography (HPLC) using poly (octadecylsiloxane) as a stationary phase, phosphate buffer as a mobile phase, a series of R-S imidazole derivatives as solutes, and beta-CD (beta-CD) and hydroxypropyl-beta-CD (HP-beta-CD) as chiral selectors, a study on the hydrophobic effect on both the solute complexation with the chiral selector and chiral discrimination mechanisms was carried out by varying the sucrose concentration c in the mobile phase and the column temperature T. An original mathematical treatment was developed to calculate the degree of complexation, rho (the percent of complexed guest solute), and the number of sucrose molecules excluded from both the uncomplexed solute-RP18 stationary phase interface (when the solute transfer occurred) and the solute-CD interface during the complexation process. This number of sucrose molecules was an image of the relative intensity of the hydrophobic effect and the inclusion degree of the solute in the chiral selector. Different Van't Hoff plot shapes of the degree of complexation and sucrose molecule number were observed with beta-CD and HP-beta-CD indicating a change in the solute inclusion and chiral discrimination. PMID- 11924512 TI - [Wheat gliadin nanoparticle size control by determination of the solubility parameters of these plant proteins]. AB - Gliadins nanoparticles, protein complex from wheat gluten, were prepared by coacervation (macromolecules desolvatation). The objective was to prepare nanometric colloidal systems whith both smallest particle size and polidispersity index. Macromolecules solvent choice is predominant. Their total solvatation enables of control the desolvatation process and thence particle sizes. Different pharmaceutical solvents were assayed. In order to select the best of them, a thermodynamical approach was used. It rests on the determination of the solubility parameter delta G of the gliadins complex. The best solvent is the one whose solubility parameter (calculated or experimentally determined) is equal to the plant protein's. PMID- 11924513 TI - [Saponins and hemolytic activity. Saponins and glycosides from five species of Sapindaceae]. AB - The haemolytic activity of saponins has been a known phenomenon for many years. In this work, structure-activity relationships were established from various triterpene saponins distinguishing hemolytic activity between mono- and bidesmosidic saponins. Saponins structure in Sapindaceae serve as a chimiotaxonomic criteria for dividing the species into two sub-families. In order to verify this criteria, five species were studied: Smelophyllum capense and Dimocarpus fumatus from the sub-family Sapindoideae, Filicium decipiens, Hippobromus pauciflorus, and Harpullia cupanioides from the sub-family Dodonaeoideae. Structure of eleven new saponins were elucidated from four species by the study of homo- and heteronuclear advanced NMR experiments and MS. Other metabolites were also isolated from Dimocarpus fumatus, including isoprenypchromenes and three new glycosides of long chain fatty alcohols. PMID- 11924514 TI - [Chemistry of cosmetics in antiquity]. AB - Several texts, statues and paintings denote the importance of make up and eye medicines since the earliest periods of Egyptian history. We have investigated cosmetic powders that were preserved in original alabaster and reed containers. Quantitative crystallographic and chemical analysis of the mineral and organic components revealed surprising facts. In addition to the well known galena PbS and cerussite PbCO3, two unexpected constituents have been identified: laurionite PbOHCl and phosgenite Pb2 (CO3) Cl2, which are rare halide minerals found in lead slag only in certain places where the sea water has weathered lead debris left over from silver mining operations in Antiquity. Alteration of natural lead minerals is also unlikely, given the excellent state of conservation of the reed vessels. This evidence indicates that laurionite and phosgenite were synthesised artificially. Support for this statement comes from recipes of medicinal products to be "used in ophthalmology" reported by Greco-Roman authors such as Dioscorides and Pline (1st Century B.C.): silver foam PbO is crushed and mixed with rock salt and sometimes with natron (Na2CO3). The reaction seems to be straightforward. However, our experiments in the laboratory have shown a major difficulty, arising from the concomitant production of alkali, which raises the pH and leads to different products. It follows that the Egyptians very early mastered this kind of chemical synthesis and technology, a fact of great importance in the History of Sciences. Fire-based technology had been mastered to manufacture Egyptian Blue pigments since the third millennium B.C. The present results now suggest that wet chemistry was already known 4000 years ago. This key finding provides a new insight into the chemical technology of far greater antiquity than has previously been believed. Yet, an important question remains relative to the ultimate motivation for these technological developments. If the Egyptians initially only aimed at different nuances of grey, they could use cerussite, an easily accessible mineral, crushed and mixed with galena. Thus, one should seek other motivations; we shall propose to consider a religious-ritual context, ultimately linked to a pharmaceutical aspect. PMID- 11924515 TI - [Patentability of the genome: what's the strategy for France?]. PMID- 11924516 TI - HSDM and the world' most populous country. PMID- 11924518 TI - Medical missions mend fences in Vietnam. PMID- 11924517 TI - Honduras diary. PMID- 11924519 TI - Heart failure satellite meeting--focus on new and evolving strategies. PMID- 11924521 TI - PROGRESS will change the way we view stroke treatment. PMID- 11924520 TI - Weight loss can be as simple as 10, 20, 30--Knoll launches blueprint for the use of sibutramine in weight management. PMID- 11924522 TI - New insights from PCI-CURE study. PMID- 11924523 TI - Papers from the Twenty-first Meeting of the International Society for Clinical Biostatistics. Trent, Italy, September 4-8, 2000. PMID- 11924524 TI - [Medical-economic evaluation of percutaneous transluminal angioplasty and aortocoronary bypass in the management of coronary artery disease (excluding myocardial infarction). December 2000]. PMID- 11924525 TI - Gene therapy with apoptin induces regression of xenografted human hepatomas. AB - The chicken anemia virus-derived Apoptin protein shows remarkable specificity; namely, it induces apoptosis in tumor cells, but not in normal diploid cells. We have exploited the Apoptin gene for use in cancer gene therapy. Here we demonstrate that adenovirus-mediated intratumoral transfer and expression of the Apoptin gene results in regression or complete remission of human hepatomas grown as xenografts in immune-deficient mice, and significantly increases their survival long term. Early after intratumoral injection, Apoptin could be detected in significant quantities by Western blot analyses and immunohistochemistry. Furthermore, cell death and disruption of the tumor integrity were apparent in the transduced regions. This experimental gene therapeutic strategy constitutes a unique example of specific antitumor activity using a virus-derived gene with broad-spectrum applicability. PMID- 11924526 TI - Inhibition of septic shock in mice by an oligopeptide from the beta-chain of human chorionic gonadotrophin hormone. AB - Human chorionic gonadotrophin (hCG) is a heterodimeric placental glycoprotein hormone required in pregnancy. In human pregnancy urine and in commercial hCG preparations (c-hCG) it occurs in a variety of forms, including breakdown products. Several reports have suggested modulation of the immune system by intact hormone, but such effects of breakdown products have not been reported. In a related article (Hum Immunol 62:1315, 2001), it is reported that a 400-2000 Dalton (Da) fraction from c-hCG and from human pregnancy urine inhibits Th1 mediated diabetes in NOD mice. The active component(s) were called natural (immuno)modulatory pregnancy factor(s) (NMPF). This study reports that a single treatment with the same low molecular weight NMPF fraction up to 24-h after high dose lipopolysaccharide (LPS) injection inhibited septic shock in mice. This counteracting effect of NMPF paralleled the downregulation of the effects of LPS on the production of macrophage migration inhibitory factor (MIF) by spleen cells, on the plasma level of liver aminotransferase, and on the expression of several splenic lymphocyte and macrophage surface markers. Based on the primary structure of the beta-chain of hCG a synthetic hexapeptide Valine-Leucin-Proline Alanine-Leucine-Proline (VLPALP) was designed, which demonstrated it to have the same protective effects as the 400-2000 Da NMPF fraction. These results indicate a new strategy for the treatment of septic shock and the potential of therapeutic use of this synthetic oligopeptide. PMID- 11924527 TI - Polymorphisms at the TNF locus in Chinese Han population. AB - One hundred sixty-four unrelated healthy individuals from Chinese Han population were investigated in order to define the distribution of eight polymorphic loci within the tumor necrosis factor (TNF) gene cluster and determine their relationship between the high polymorphic microsatellite TNFa, b, d, and other elements. The cloning and sequencing for five microsatellites were simultaneously done. In this study, the distribution of TNF alleles apparently vary from other ethnic groups. A new allele was detected and confirmed. It should be emphasized that a very strong association between TNFd8 and TNFe4 is reported and d8e4 haplotype appears to be specific to the population studied. In addition, five extended haplotypes were established in this population: a6b5c1d8e4TNF308-1TNF betaNco1-1TNFAspH1-2, a2b1c2d5e1TNF308-1TNF-betaNco1-2TNFAspH1-2, a11b4c1d4e3TNF308-1TNF-betaNco1-2TNFAspH1-1, a10b4c1d4e3TNF308-1TNF-betaNco1 2TNFAspH1-1, and a2b3c1d2e3TNF308-2TNFAspH1-2. Data suggest that important ethnic differences may exist and that it is a necessary initiative for further research. PMID- 11924528 TI - A practice is the business of the professional person (what they didn't tell you about business in dental school). PMID- 11924529 TI - Effect of age on thrombosis and morbidity in the mouse-limb transplantation model. AB - Murine models of limb transplantation have been recently described. Because of the technical challenge, non-technical factors that influence the success rate and are easily controlled should be considered. This study investigated the effect of recipient animal age on anastomotic thrombosis, morbidity, and mortality. Twelve allogeneic heterotopic hindlimb transplants were performed using the femoral vessels with end-to-end arterial and end-to-side venous anastomoses. Group 1 (n=8) consisted of 2 to 3-month-old mice weighing 17 to 20 g, and Group 2 (n=7) included 7 to 8-month-old mice weighing 24 to 27 g. In Group 1, 6/8 (75 percent) transplants were successful, while in Group 2, only 1/7 (14 percent) allografts survived (p<0.05). A statistically significant difference in the incidence of vascular compromise of the ipsilateral recipient hindlimb was also noted (p<0.01). The authors conclude that although smaller, juvenile, inbred mice have a higher anastomotic patency rate, with greater collateral vasculature in the hindlimbs, and are therefore more suitable for limb transplantation research. PMID- 11924530 TI - [Editorial]. PMID- 11924531 TI - Exon/intron organisation of human proteasome PROS-27 K gene. AB - The exon/intron structure of the human proteasome PROS-27 gene was established by means of partial sequencing of its genomic clones and comparison with the chromosome 14 sequences from the data bases. The gene contains seven exons spanning over 19kb. Introns of the gene contain numerous Alu type repeats, Mer 2 and LINE type repeats. Pattern of the repeats indicates conservatism of the sequence. PMID- 11924532 TI - Sequencing and chromosomal assignment of the rat endothelial-derived lipase gene (Lipg). AB - Part of the nucleotide sequence of the Lipg gene in the rat was established using primers based on the mRNA sequence described in the mouse. The rat intron sequence served as a template for designing primers for the specific amplification of rat Lipg. A rat-hamster radiation hybrid (RH) panel was used for chromosomal assignment of the rat Lipg gene. The Lipg gene was found to be located on rat chromosome 18 in the vicinity of the marker D18Mit11; a region reported to be homologous with both human and mouse chromosome 18. PMID- 11924533 TI - Effects of low doses of inhaled nitric oxide combined with oxygen for the evaluation of pulmonary vascular reactivity in patients with pulmonary hypertension. AB - The purpose of this study was to evaluate the hemodynamic effects of inhaled nitric oxide in oxygen (NO + O2) in patients with pulmonary hypertension. Eighteen patients (median age 31.5 months) with pulmonary hypertension inhaled through a mask 100% O2 and 20 parts per million NO + inspired O2 fraction (FiO2) at 0.4. Hemodynamic measurements were made at baseline and after O2 and NO + O2 administration. The pulmonary vascular resistance index decreased after inhalation of O2 and NO + O2 (p = 0.0018 and p = 0.0003, respectively), the decrease being significantly greater after NO + O2 (p = 0.0311). Concerning the transpulmonary pressure gradient, a reduction occurred in values after O2 and NO + O2 inhalation when compared with baseline values (p = 0.0014 and p = 0.0008). In patients with congenital heart disease, an increase occurred in pulmonary blood flow after O2 (p = 0.0089) and NO + O2 (p = 0.0019) compared with baseline values, and an increase also occurred in the pulmonary/systemic blood flow ratio after NO + O2 (p = 0.0017). The main side effect related to NO + O2 was pulmonary congestion in 3 patients. Low doses of NO combined with O2 demonstrated a selective pulmonary vasodilator response in patients with pulmonary hypertension. Despite its use for testing pulmonary reactivity, inhalation of NO + O2 should be carefully administered because of the potential risk of pulmonary congestion. PMID- 11924534 TI - Does chronic pacing affect exercise capacity after Mustard operation for transposition of the great arteries? AB - Late survival after Mustard repair of transposition of the great arteries is generally good but is often characterized by progressive deterioration of ventricular function and by late postsurgical arrhythmias, thus imposing the need for permanent pacing. To evaluate how chronic pacing affects long-term exercise capacity, we compared two groups of these patients: group 1, comprising 12 patients, aged 9.0 +/- 2.6 years, without pacemaker; and group 2, comprising 18 patients, aged 9.3 +/- 2.0 years, with pacemaker. Patient evaluation included history, physical examination electrocardiograph, Holter monitoring, and echocardiography. Pacing modes were as follows: AAI (6 patients), AAIR (9 patients), VVI (2 patients), and VVIR (1 patient). At exercise test we evaluated exercise tolerance, maximum heart rate, blood pressure, oxygen consumption, and cardiac output at rest and at peak exercise. The two groups were comparable for all variables examinated. All pacemakers showed normal function. During the exercise, 11 of 12 patients in group 1 showed sinus rhythm, and in group 2, 11 patients showed sinus rhythm, 5 junctional rhythm, and 2 continuous pacing. There were no significant differences between groups. Chronically paced Mustard patients but with restoration of spontaneous rhythm during the exercise test do not show reduced exercise tolerance in comparison with nonpaced Mustard patients. PMID- 11924535 TI - Plastic bronchitis in children with Fontan palliation: analogue to protein losing enteropathy? AB - We studied a 6-year-old boy and a 2-year-old girl with bronchitis fibroplastica following Fontan operation. Large endobronchial casts of rubber-like consistency resulted in life-threatening pulmonary failure. In one patient symptoms improved after optimizing heart function with diuretics, and in the other a dramatic improvement with the resolution of the clinical symptoms and normalized serum albumin followed subcutaneous high-molecular-weight heparin treatment. The severe relapse after discontinuation of the heparin medication and the once more successful treatment with heparin suggest that in addition to optimizing heart function, high-molecular-weight heparin might be a therapeutic option for this poorly understood condition. PMID- 11924536 TI - Clinical evaluation of changes in the morphology of eosinophils. AB - In order to diagnose and monitor patients with allergic diseases, we studied parameters which reflect not only the amount of eosinophils but also their state of activation. Morphologic features reveal additional information on the activity state of eosinophils. The number of nuclear lobes, cell size, amount of vacuoles and density of specific granules are considered characteristic features of eosinophils. We conclude that the size of eosinophils is a useful parameter to distinguish patients with and without eosinophilia. The treatment with corticosteroids did not affect morphological characteristics such as the number of vacuoles, granulation density, cell diameter and the nucleus/cell surface ratio, but the number of lobes per nucleus, a marker of eosinophil maturation, decreased significantly. PMID- 11924537 TI - Relationships between serum markers of monocyte/macrophage activation in type 1 Gaucher's disease. AB - We studied 44 patients with type 1 Gaucher's disease (16 non-treated patients and 28 treated with enzyme replacement therapy). We measured serum levels of chitotriosidase (ChT), neopterin, angiotensin-converting enzyme (ACE), adenosine deaminase (ADA) and beta-hexosaminidase (Hex) and its major isoenzymes Hex A and Hex B. In the untreated group of patients, the increase in serum levels was ChT>neopterin>ACE> ADA>Hex, with all decreasing significantly in treated patients (p< 0.001). Highly significant correlations were obtained between the markers of monocyte/macrophage activation which were tested (p<0.001). However, partial correlations between serum Hex B (with Hex A constant) and ChT, ACE, neopterin and ADA did not reach statistical significance. This suggests that hepatocytes are the major cellular source of this isoenzyme. Similarly, partial correlation of ChT with neopterin, with the other variables constant, was not significant, which would suggest a different expression of these two markers in Gaucher's disease. PMID- 11924538 TI - Effects of (pre-)analytical variables on activated protein C resistance determined via a thrombin generation-based assay. AB - The normalized activated protein C sensitivity ratio (nAPC-sr) determined with an assay that quantifies the effect of APC on thrombin formation initiated via the extrinsic coagulation pathway identifies hereditary and acquired defects of the protein C system. We investigated the influence of assay conditions (analytical variables) and plasma handling (pre-analytical variables) on nAPC-sr obtained with this APC resistance test. The effect of the analytical variables (CaCl2, phospholipid and APC concentrations and the concentration and source of tissue factor) was determined in pooled normal plasma. Inhibition of thrombin formation by APC was dependent on the APC concentration and was also affected by the tissue factor, Ca2+ and phospholipid concentrations. Thus, strict standardization of reactant concentrations is required to obtain reproducible nAPC-sr. Three different tissue factor preparations were compared by determining nAPCsr in plasma samples obtained from 90 healthy individuals. nAPC-sr were similar for all three tissue factor preparations although, compared with the noncommercially available tissue factor used in earlier studies, values determined with commercial tissue factor preparations showed larger variation. Pre-analytical variables, investigated in plasma of nine volunteers (3 normal individuals and 6 individuals with an APC-resistant phenotype) were: concentration of anticoagulant (3.2% vs. 3.8% trisodiumcitrate), time before processing of blood (0, 4 and 24 h), centrifugation speed, storage temperature of plasma (-20 degrees C vs. -80 degrees C) and sample thawing. Multiple linear regression analysis showed that only the citrate concentration affected the nAPC-sr, which was higher in samples collected in 3.2% trisodiumcitrate than in samples collected in 3.8% trisodiumcitrate. PMID- 11924539 TI - Cavanagh's syndrome (congenital thenar hypoplasia). AB - Thenar hypoplasia can be an isolated defect, as in Cavanagh's syndrome, can be present with cardiac (Holt-Oram syndrome) or eye (Okihiro's syndrome) disorders, or can be associated with hand anomaly, as in Haas's malformation. Vascular abnormality may be associated with thenar hypoplasia, which has been demonstrated in Okihiro's syndrome. Cavanagh's syndrome is a rare anomaly of the upper extremities that presents with unilateral or bilateral hypoplasia of the thenar eminence. Typical clinical, radiographic, and electrophysiologic findings emphasize the diagnosis. Differentiation from carpal tunnel syndrome is important to prevent unnecessary intervention. Electrophysiologic and radiographic findings are necessary tools for the physician to establish a correct diagnosis and make an appropriate referral. Because of its rarity, we present the case of an 8-year old girl with this syndrome. PMID- 11924540 TI - The impact of error training and individual differences on training outcomes: an attribute-treatment interaction perspective. AB - The authors examined the effectiveness of error training for trainees with different levels of cognitive ability, openness to experience, or conscientiousness. Participants (N = 181) were randomly assigned to control, error-encouragement, or error-avoidance conditions and trained to perform a decision-making simulation. Declarative knowledge, task performance, and self efficacy were measured posttraining. Findings suggest the effectiveness of error training is dependent on the cognitive ability or dispositional traits of trainees. High cognitive ability or more open individuals benefit more from error encouragement training than low cognitive ability or less open individuals. Conscientiousness has a negative effect on self-efficacy when trainees are encouraged to make errors. PMID- 11924541 TI - The intoxicated witness: effects of alcohol on identification accuracy from showups. AB - The effect of alcohol on identification accuracy is potentially an important topic. This study examined the effects of alcohol consumption on identification accuracy from showups, the identification procedure most likely to be used by police with intoxicated witnesses. The blood alcohol level of people exposed to a target was measured. In the target-present showup condition, blood alcohol level was not significantly related to correct identification rate. In the target absent showup condition, the higher the blood alcohol level, the more people were likely to make a false identification. Implications for law enforcement and future research directions are discussed. PMID- 11924542 TI - Responses to formal performance appraisal feedback: the role of negative affectivity. AB - This study examined the effects of performance appraisal feedback on job and organizational attitudes of tellers (N = 329) in a large international bank. Negative affectivity moderated the link between favorable appraisal feedback and job attitudes. Among the higher rated performers, attitudes were improved 1 month after being notified of favorable appraisal results (Time 2). Improved attitudes persisted 6 months after the performance appraisal (Time 3) among tellers with low negative affectivity but not among those with high negative affectivity. Among the lower rated performers, mean levels of attitudes did not change significantly during the study. PMID- 11924543 TI - UV absorbance dependent toxicity of acridine to the marine diatom Phaeodactylum tricornutum. AB - The present study seeks quantitative measures for photoenhanced toxicity under natural light regimes by comparing the effects of an aromatic compound under natural and laboratory light. To this purpose, the influence of light irradiance and spectral composition on the extent of photoenhanced toxicity of acridine, a three-ringed azaarene, to the marine diatom Phaeodactylum tricornutum was analyzed. Under laboratory light containing ultraviolet radiation (UV), the 72-h EC50 growth value for acridine was 1.55 microM. Under natural light, a 72-h EC50 value for acridine below the lowest test concentration (0.44 microM) was observed. Under both laboratory and natural light, the toxicity of acridine was equally enhanced by total UV (UV-A and UV-B) and UV-A radiation, while in the absence of UV no enhancement of toxicity was observed. Hence, the UV-A region of light was dominant in the photoenhanced toxicity of acridine to P. tricornutum, in accordance with its absorption spectrum in the UV-A region. Therefore, the total amount of UV radiation absorbed by aqueous acridine was calculated for each separate treatment. The amount of UV absorbed by acridine effectively described the effect of acridine on the growth of P. tricornutum in a dose-response dependent manner. It is concluded that photoenhanced toxicity of aromatic compounds expressed as a function of the actually absorbed UV may circumvent some of the variability between studies using different concentrations of the phototoxic compounds and light sources. The UV quantity absorbed by these compounds allows a comparison with the absorption characteristics of natural waters and, thus, is a key parameter to determine the role of photoenhanced toxicity in water. PMID- 11924544 TI - Predicting arsenic solubility in contaminated soils using isotopic dilution techniques. AB - An isotopic dilution assay was developed to measure radiolabile As concentration in a diverse range of soils (pH 3.30-7.62; % C = 1.00-6.55). Soils amended with 50 mg of As kg(-1) (as Na2HAsO4 x 7H2O) were incubated for over 800 d in an aerated "microcosm" experiment. After 818 d, radiolabile As ranged from 27 to 57% of total applied As and showed a pH-dependent increase above pH 6. The radiolabile assay was also applied to three sets of soils historically contaminated with sewage sludge or mine-spoil. Results reflected the various geochemical forms in which the arsenic was present. On soils from a sewage disposal facility, radiolabile arsenate ranged from 3 to 60% of total As; mean lability was lower than in the equivalent pH range of the microcosm soils, suggesting occlusion of As into calcium phosphate compounds in the sludge-amended soils. In soils from mining areas in the U.K. and Malaysia, radiolabile As accounted for 0.44-19% of total As. The lowest levels of lability were associated with extremely large As concentrations, up to 17,000 mg kg(-1), from arsenopyrite. Soil pore water was extracted from the microcosm experiment and speciated using "GEOCHEM". The solid<==>solution equilibria of As in the microcosm soils was described by a simple model based on competition between HAsO4(2-) and HPO4(2-) for "labile" adsorption sites. PMID- 11924545 TI - A mass balance model describing multiyear fate of organochlorine compounds in a high Arctic lake. AB - Data collected over a 3-year study of a high arctic watershed and lake are used to understand the fate of organochlorine compounds (OCs) and form the basis of a mass balance contaminant fate model. The model uses the fugacity/aquivalence approach to describe OC dynamics between air, stream inflows and outflow, the water column, and surficial sediments. The steady-state model results indicate that stream inflows contributed from 96 to >99% of total chemical loadings, but 57-98% of total loadings were lost from the lake via the outlet, the percentage of which is controlled by the hydrologic regime of the high arctic lake. Conversely, only 0.4-3.4% of loadings were retained within the sediments due to the high export rate, minimal scavenging from the water column and low organic carbon fraction of the sediments. Using the unsteady-state model, which includes year-round processes, degradation was estimated to account for losses of 7-32% for the more persistent OCs and 42-50% for the less persistent OCs (alpha-HCH, gamma-HCH, and endosulfan I). If loadings were eliminated, water column concentrations would decline with half-lives <1 year for less persistent OCs and 1-2 years for the more persistent OCs, whereas the half-lives for OCs in sediment are 8-25 years. PMID- 11924546 TI - 1H and 11B NMR study of p-toluene boronic acid and anhydride. PMID- 11924547 TI - Need for caesarean sections in west Africa. PMID- 11924548 TI - Distribution study of 3,4-methylenedioxymethamphetamine and 3,4 methylenedioxyamphetamine in a fatal overdose. AB - In this study, regional tissue distributions of the amphetamine analogue 3,4 methylenedioxymethamphetamine (MDMA, "ecstasy") and its metabolite 3,4 methylenedioxyamphetamine (MDA) in a fatal overdose are presented. Quantitation of MDMA and MDA levels occurred in blood samples taken centrally (right and left heart and main adjacent great vessels) and peripherally (subclavian and femoral blood). In addition, MDMA and MDA concentrations were determined in cardiac and iliopsoas muscle, both lungs, liver, both kidneys, spleen, the four brain lobes, cerebellum and brainstem, and adipose tissue. Finally, MDMA and MDA levels were determined in serum, vitreous humor, urine, and bile. For all samples, a fully validated high-pressure liquid chromatography procedure with fluorescence detection was used. The found substances were also identified with liquid chromatography-tandem mass spectrometry. Our data confirm that blood sampling from an isolated peripheral vein is recommended for MDMA and MDA. In addition, the vitreous humor MDMA level indicates that this fluid can be an interesting alternative when a suitable blood sample is missing. Considering the substantial differences in concentrations in blood samples taken from various sites in the body and the high levels in some tissues (e.g., in liver), we concluded that the influence of postmortem redistribution should be taken into account in the interpretation of toxicological data when an appropriate peripheral sample cannot be obtained or when blood samples are not available because of putrefaction. PMID- 11924549 TI - Impact of wildfires on the air quality of Mexico City, 1992-1999. AB - Wildfires in Mexico increased in 1998, compared to information for the last 6 years. The average number of wildfires in the Mexico City Metropolitan Area (MCMA) for this year (1998) were 58% (1916 events) more events than the 1992-1997 (average cases 1217 events). Mexico City affected area corresponds to 1.3% of the national affected area. The purpose of this paper is to evaluate the impact on the particles air quality due to the wildfire emissions at the MCMA and surrounding areas. Using the corresponding US EPA emission factors for wildfires, the tons of particulate matter, nitrogen oxides, carbon monoxide, and total hydrocarbons emitted by this source for the MCMA case were obtained. The calculated emissions during wildfires were correlated with the levels of particles present in the atmosphere. A comparison of the concentration levels of particles, both as PM10 as well as TSP, were made for the years 1992-1998, during wet and dry season, being March, April, and May the critical months due to the presence of wildfires. A good correlation is observed between particulate wildfire emissions and particulate air quality, being stronger for TSP. A clear impact on the particles air quality due to the increase of wildfires in 1998, is observed when this year is compared with 1997, presenting an increment of 200 300% for some monitoring stations. PMID- 11924550 TI - Tolerance and acclimation to zinc of Ceriodaphnia dubia. AB - Zinc is an essential metal for all living organisms. However, so far, little or no attention has been paid to the consequences of zinc deficiency or acclimation to this metal during culturing and testing on toxicity test results. In this study, the cladoceran Ceriodaphnia dubia was acclimated for 10 generations to four zinc concentrations ranging from 0 to 100 microg Zn/l and changes in zinc tolerance were monitored using acute (48 h) and chronic (9 days) assays. C. dubia deprived of zinc and acclimated to 13 microg Zn/l had a lower fitness in comparison with organisms acclimated to 50 and 100 microg Zn/l. In the two lowest versus the two highest acclimation concentrations the 9dEC50 values (on immobility) were 358-387 microg Zn/l versus 486-489 microg Zn/l; the mean number of young per female was 11-18 versus 25-32; and the time to first brood was 4.7 5.0 days versus 4.0-4.3 days. Moreover, the coefficient of variation of all parameters tested was highest in the two lowest acclimation concentrations. The results indicate that culturing test animals in media lacking trace metals such as zinc could give rise to animals that are unnaturally sensitive to those same metals daring toxicity tests. PMID- 11924552 TI - Dutch goats with a thyroglobulin mutation. PMID- 11924551 TI - Use of aminoglutethimide in the treatment of pituitary-dependent hyperadrenocorticism in the dog. AB - The aim of this study was to evaluate the efficacy and safety of aminoglutethimide in the treatment of dogs with pituitary-dependent hyperadrenocorticism (PDH). Ten dogs were diagnosed with PDH based on clinical and laboratory data, adrenal function tests (adrenocorticotropic hormone [ACTH] stimulation test and urinary cortisol/creatinine ratio [UCCR] combined with a high dose oral dexamethasone suppression test) and ultrasonographic evaluation of the adrenal glands. Aminoglutethimide was administered daily at a dose of 15 mg/kg bodyweight for one month. Median basal cortisol concentration and post-ACTH cortisol concentration one month after treatment were significantly lower than pretreatment values. Complete response was achieved in one dog, and partial response was obtained in three dogs. Severe side effects of anorexia, vomiting and weakness occurred in one dog and medication was withdrawn. Two further dogs developed decompensations of concurrent diseases and medication was stopped in these animals as well. Mild toxicity occurred in four dogs. Moderate to severe elevations in liver enzymes occurred in all dogs. The efficacy of this drug is lower than that observed using mitotane and ketoconazole, and adverse effects limit its use. Aminoglutethimide, using the protocol described, cannot be recommended for long-term management of PDH in the dog. PMID- 11924553 TI - The nursing portfolio: a reflection of a professional. AB - A nursing portfolio is a means of documenting skills such as communication, creativity, and critical thinking. More important, it reflects professional development, self-evaluation, and achievement of goals. Designing and implementing a portfolio enables traditional nursing students and RNs returning to school to describe their achievements during a specified period of time. For the adult student seeking scholastic or clinical credit for past work or life experiences, the portfolio provides a creative and structured way for this to be obtained without taking a standardized test. PMID- 11924554 TI - First confirmation of porcine colonic spirochaetosis caused by Brachyspira pilosicoli in Iberian pigs in Spain. PMID- 11924556 TI - Point and click. (If only it were that easy.). AB - Along with tight reimbursement and confounding regulations, any list of big headaches for health care leaders includes information technology. We've been promised so much from IT for so long, and have been disappointed so many times, who can blame us for being skeptical, if not downright cynical? When is the last time an investment in information technology actually improved your bottom line? But we know we can't dismiss IT altogether. At the end of the rainbow there really is efficiency, security and better care for patients. So how close are we? In this issue, H&HN takes a hard look at technology--what it can do for us today and tomorrow. We look at return on investment, examining whether hospitals are measuring what they get back against what they put in; a budding effort to bring a common platform to health care IT; handheld computers; electronic medical records; and computerized physician order entry. The technology is out there, but it's fair to ask: is it really clicking yet? PMID- 11924555 TI - Use of alternative pharmacotherapy in management of cardiovascular diseases. AB - OBJECTIVES: To review use of alternative pharmacotherapy (AP) in patients with cardiovascular disease (CVD) and significant drug interactions between AP and traditional CVD medications. STUDY DESIGN: A literature search of MEDLINE and the National Complementary and Alternative Medicine database was done using these search terms: supplements, vitamins, garlic, fish oil, L-arginine, soy, coenzyme Q10, herbs, phytosterols, chelation therapy, alternative medicine, and CVD. PATIENTS AND METHODS: English human clinical trials measuring surrogate and clinical end points. RESULTS: Antioxidants have not been consistently proven beneficial in reducing cardiovascular mortality. Fish oils may be beneficial in patients with hypertension and hypercholesterolemia, but therapeutic doses need to be defined. Use of coenzyme Q10 in patients with heart failure has not demonstrated consistent benefits. Garlic may lower blood pressure and cholesterol levels, but also may increase bleeding, so its use in CVD patients should be monitored. Clinical studies with small sample sizes have demonstrated that L arginine may be useful to prevent and treat CVD. The Food and Drug Administration recommends 25 g/day of soy protein as part of a diet low in saturated fats for cholesterol reduction. Plant sterols are recommended by the American Heart Association and the National Cholesterol Education Program Expert Panel as adjunct therapy to reduce low-density lipoprotein. No data support use of chelation therapy. Some APs interact with common prescription CVD medications (eg, gingko and ginseng with warfarin, St. John's Wort with digoxin). CONCLUSIONS: The benefits of APs as part of the treatment for CVD are controversial. Routine use is not recommended. PMID- 11924557 TI - Mutational and haplotype analysis of AGL in patients with glycogen storage disease type III. AB - Glycogen storage disease type III (GSD III) is a rare autosomal recessive inherited disorder caused by a deficiency of the glycogen-debranching enzyme (AGL). We investigated two GSD III patients and identified four different mutations. Nucleotide sequence analysis revealed patient 1 of Chinese descent to be a compound heterozygote for a novel nonsense mutation, R34X, and the splicing mutation (IVS32-12A > G) reported in a Japanese patient. Patient 2 of Japanese origin was found to be compound heterozygous for a novel nonsense mutation, Y1148X, and the splicing mutation (IVS14+1G > T) that we had described previously. To determine whether splicing mutations occurred independently, we performed intense AGL haplotype analysis using 21 intragenic polymorphic markers plus a novel polymorphism IVS32-97 A/G in the vicinity of the IVS32 splicing mutation. Patient 1 of Chinese origin and the Japanese patient homozygous for the IVS32-12A > G were found to have different haplotypes, indicating the IVS32-12A > G mutation to be a recurrent mutation. This is the first recurrent mutation established by intense haplotyping in the AGL gene. PMID- 11924558 TI - [Diseases of food origin]. PMID- 11924559 TI - Disease incidence and immunological traits for the selection of healthy pigs. A review. AB - Disease is a major issue in animal production systems and society demands that the use of medicines and vaccines be reduced. This review describes the breeding approaches that could be used to improve disease resistance and focuses especially on their application to pigs. Disease reduction by genetic means has certain advantages through cumulative and permanent effects, and direct and indirect selection methods are available. Direct selection for disease incidence requires, besides a unique pig identification and disease registration system, challenge routines that are inconvenient in intensive pig production. Indirect selection for the expression of immune capacity may be an alternative but requires detailed knowledge of the different components of the immune system. There is ample opportunity for genetic improvement of the immune capacity because immune traits show substantial genetic variation between pigs. We therefore conclude that indirect selection via immune traits is very interesting, also for practical implementation, and that there is an urgent need for knowledge, within lines, about the genetic relationships between immune capacity traits and resistance to specific diseases or to disease incidence in general. Furthermore, knowledge about the relationship between immune system traits and production traits is needed as well as knowledge about the effect of selection on the epidemiology of disease at a farm/population level and on the host-pathogen interaction and coevolution. PMID- 11924560 TI - The Dutch Brucella abortus monitoring programme for cattle: the impact of false positive serological reactions and comparison of serological tests. AB - The Dutch national Brucella abortus eradication programme for cattle started in 1959. Sporadic cases occurred yearly until 1995; the last infected herd was culled in 1996. In August 1999 the Netherlands was declared officially free of bovine brucellosis by the European Union. Before 1999, the programme to monitor the official Brucella-free status of bovine herds was primarily based on periodical testing of dairy herds with the milk ring test (MRT) and serological testing of all animals older than 1 year of age from non-dairy herds, using the micro-agglutination test (MAT) as screening test. In addition, serum samples of cattle that aborted were tested with the MAT. The high number of false positive reactions in both tests and the serum agglutination test (SAT) and complement fixation test (CFT) used for confirmation seemed to result in unnecessary blockade of herds, subsequent testing and slaughter of animals. For this reason, a validation study was performed in which three indirect enzyme-linked immunosorbent assays (ELISAs), the CFT and the SAT were compared using a panel of sera from brucellosis-free cattle, sera from experimentally infected cattle, and sera from cattle experimentally infected with bacteria which are known to induce cross-reactive antibodies (Pasteurella, Salmonella, Yersinia, and Escherichia). Moreover, four ELISAs and the MRT were compared using a panel of 1000 bulk milk samples from Brucella-free herds and 12 milk samples from Brucella abortus- infected cattle. It is concluded that the ELISA obtained from ID-Lelystad is the most suitable test to monitor the brucelosis free status of herds because it gives rise to fewer false-positive reactions than the SAT. PMID- 11924561 TI - Muscarinic cholinergic receptor blockade impairs free fatty acid mobilization during fasting in pigeons (Columba livia). AB - The effects of intracerebroventricular pretreatment with muscarinic (scopolamine or methylscopolamine; 2.7 nmol or 5.4 nmol) or nicotinic (mecamylamine, 2.7 nmol or 5.4 nmol) cholinergic receptor antagonists on plasma free fatty acid increases induced by intracerebroventricular injections of carbachol in conscious resting pigeons (Columba livia) were examined. Plasma glucose levels were also measured throughout the experiments. Pretreatment with methylscopolamine suppressed the lipolytic effect of carbachol injections, while mecamylamine left this response unchanged. Neither carbachol treatment alone, nor the pretreatments with cholinergic agents affected glucose levels. Subsequently, the effects of intracerebroventricular injections of methylscopolamine were investigated in 24-h food-deprived pigeons. The increase in free fatty acid levels after fasting was of a magnitude similar to that observed after carbachol treatment; intracerebroventricular injections of methylscopolamine (5.4 nmol) transiently but powerfully decreased plasma free fatty acids in 24-h food-deprived pigeons to levels comparable to those of free-feeding animals. The fasting-induced decrease in glucose levels was not affected by this treatment. These data indicate that the lipolytic response induced by carbachol may be mediated by central muscarinic cholinergic receptors and that this central cholinergic mechanism partially contributes to plasma free fatty acid increases observed during fasting. Furthermore, the absence of effects on glucose levels suggests that these cholinergic mechanisms participate selectively in the lipolytic component of the metabolic response to fasting. PMID- 11924562 TI - Excitation-contraction coupling in cardiac muscle of lobster (Homarus americanus); the role of the sarcolemma and sarcoplasmic reticulum. AB - The T-tubules and sarcoplasmic reticulum (SR) serving excitation-contraction (EC) coupling in lobster (Homarus americanus) cardiac muscle are similar to those in mammalian myocardium. Tetanic contraction is elicited by a burst of action potentials from the cardiac ganglion. In this study we evaluated the roles of the sarcolemma and SR in EC coupling of the ostial valve muscle (orbicularis ostii m. or OOM) of lobster heart. The OOM was mounted in a bath with saline on a microscope stage; force was measured by strain gauge. [Ca2+]i was measured using iontophoretically micro-injected fura-2 salt. Peak [Ca+]i, peak tetanic force and time to peak [Ca2+]i increased with that of stimulus train duration (TD), to a maximum at a TD of 500 ms. Force increased with [Ca2+]. Cd2+ reduced force by 90%; ryanodine and caffeine reduced tetanic [Ca2+]i transients by 80% and 70%, and force by 90% and 80%, respectively. Ryanodine, caffeine and cyclopiazonic acid slowed the decline of [Ca2+]i and force during relaxation. Relaxation required [Na+]o. The rate of decline of [Ca2+]i appeared to be a sigmoidal function of the [Ca2+]i and increased for any [Ca2+]i with TD. Inactivity slowed relaxation of force; stimulation accelerated relaxation. These data suggest important contributions of Ca2+ transport both across the sarcolemma and across the SR membrane during EC-coupling of lobster cardiac muscle, while average cytosolic [Ca2+]i regulates the rate of [Ca2+]i elimination during relaxation. PMID- 11924563 TI - Simple roost nests confer large energetic savings for sparrow-weavers. AB - White-browed sparrow-weavers (Plocepasser mahali, body mass 40 g) are group living passerines adapted to the semi-arid environment of north-eastern and south western Africa. During winter, the nocturnal ambient temperature of these regions often falls below 0 degrees C. imposing conditions demanding an increase in thermoregulatory heat production. Individuals roost throughout the year in inverted U-shaped roost nests. We investigated the energetic advantages of roosting by measuring nest and ambient temperatures in the field, as well as the resting metabolic rate (RMR) of the birds. The sparrow-weavers exhibited a wide thermoneutral zone (13 degrees C - 32 degrees C). Although RMR at thermoneutrality (40.2 J g.h(-1)) conforms with those of other passerines. the value at 0 degrees C (74.8 J g.h(-1)) is significantly lower than expected. The slope of the line below the lower critical temperature is unexpectedly steep, however, and appears to cause the physiological requirement for nest roosting due to a high cost of thermoregulation at low temperatures, perhaps due to shivering or non-shivering thermogenesis. The nest temperature at 0 degrees C ambient is 5 degrees C. resulting in a saving of some 7% in the energy spent during winter nights when food resources are in short supply compared with the rest of the year. PMID- 11924564 TI - CQI 'business as usual' is no help for patient safety. PMID- 11924565 TI - Patient notes: transient ischemic attacks. PMID- 11924566 TI - [Frequency of vaginosis caused by Gardnerella vaginalis]. PMID- 11924567 TI - The 5' and 3' extremities of the satellite tobacco necrosis virus translational enhancer domain contribute differentially to stimulation of translation. AB - The translational enhancer domain (TED) of satellite tobacco necrosis virus (STNV) RNA stimulates translation of uncapped RNAs autonomously. Here we set out to identify the 5' and 3' extremities of TED and features of these sequences with respect to translation. We found that both in wheat germ extract and in tobacco protoplasts, the 5' border is confined to 3 nt. Mutational analysis revealed that the autonomous function of TED is sensitive to 5' flanking sequences. At the 3' end of TED, 23 nt have a cumulative, quantitative effect on translation in wheat germ extract, whereas in tobacco protoplasts, the most 3' 14 nt of these 23 nt do not enhance translation. The 5' and 3' sequence requirements triggered the development of a new secondary structure model. In this model, TED folds into a phylogenetically conserved stem-loop structure in which the essential 5' nucleotides base-pair with the 3' nucleotides that stimulate translation both in vitro and in vivo. Importantly, the 14 3' nucleotides in TED that stimulate translation in the wheat germ extract only do not require the predicted base pairing in order to function. The discrepancy between in vitro and in vivo sequence requirements thus correlates with potential base-pairing requirements, opening the possibility that TED contains two functional domains. PMID- 11924568 TI - Use of alternative medicine by patients in a rural family practice clinic. AB - BACKGROUND AND OBJECTIVES: There has been an increasing awareness of the use of alternative medicine and its effect on health care in the United States. However, no previous study has looked at its use among primary care patients in a rural setting. We conducted this study to determine the patterns of use of alternative medicine in this population. METHODS: A questionnaire was distributed to 750 adult patients in a family practice clinic in northern Pennsylvania. RESULTS: Our response rate was 88% (664/750). Forty-seven percent of patients reported using at least one form of alternative medicine during the past year The most-common types used were chiropractic (used by 17.2% of respondents), relaxation techniques (16.9%), herbal medicine (16.9%), and massage (14.2%). The patients surveyed used alternative medicine more for its benefits than because of dissatisfaction with conventional medicine. Only 51% of patients told their physician about their use of alternative medicine. CONCLUSIONS: A significant number of rural family practice patients are using alternative medicine. To better address their patients' needs, primary care physicians should routinely ask patients about their use of alternative medicine and advise them accordingly. PMID- 11924569 TI - Overcoming barriers to the early detection and treatment of chronic kidney disease and improving outcomes for end-stage renal disease. AB - Chronic kidney disease (CKD) is a growing public health problem. Morbidity and mortality rates from CKD and end-stage renal disease remain high despite the advent of new knowledge, therapies, and clinical practice guidelines. Consequently, the reasons for the underdiagnosis and undertreatment of CKD must be addressed and appropriate preventive and therapeutic interventions implemented. Managed care organizations (MCOs) can facilitate this process by implementing intervention programs that target clinicians and patients with kidney disease. These programs should promote 1) early detection of CKD and comorbid conditions; 2) use of appropriate outcome measures to stratify patient care; 3) implementation of strategies to delay disease progression and prevent or treat complications; and 4) adequate preparation for and timely initiation of renal replacement therapy. Early detection and proactive care of predialysis patients not only improve outcomes and quality of life, but also reduce costs for society and MCOs. PMID- 11924570 TI - Invariant recognition of feature combinations in the visual system. AB - The operation of a hierarchical competitive network model (VisNet) of invariance learning in the visual system is investigated to determine how this class of architecture can solve problems that require the spatial binding of features. First, we show that VisNet neurons can be trained to provide transform-invariant discriminative responses to stimuli which are composed of the same basic alphabet of features, where no single stimulus contains a unique feature not shared by any other stimulus. The investigation shows that the network can discriminate stimuli consisting of sets of features which are subsets or supersets of each other. Second, a key feature-binding issue we address is how invariant representations of low-order combinations of features in the early layers of the visual system are able to uniquely specify the correct spatial arrangement of features in the overall stimulus and ensure correct stimulus identification in the output layer. We show that output layer neurons can learn new stimuli if the lower layers are trained solely through exposure to simpler feature combinations from which the new stimuli are composed. Moreover, we show that after training on the low-order feature combinations which are common to many objects, this architecture can- after training with a whole stimulus in some locations--generalise correctly to the same stimulus when it is shown in a new location. We conclude that this type of hierarchical model can solve feature-binding problems to produce correct invariant identification of whole stimuli. PMID- 11924571 TI - Osteoporosis and bone marrow mastocytosis: dissociation of skeletal responses and mast cell activity during long-term bisphosphonate therapy. PMID- 11924572 TI - Human osteoblasts' proliferative responses to strain and 17beta-estradiol are mediated by the estrogen receptor and the receptor for insulin-like growth factor I. AB - The mechanism by which mechanical strain and estrogen stimulate bone cell proliferation was investigated using monolayer cultures of human osteoblastic TE85 cells and female human primary (first-passage) osteoblasts (fHOBs). Both cell types showed small but statistically significant dose-dependent increases in [3H]thymidine incorporation in response to 17beta-estradiol and to a single 10 minute period of uniaxial cyclic strain (1 Hz). In both cell types, the peak response to 17beta-estradiol occurred at 10(-8) - 10(-7) M and the peak response to strain occurred at 3500 microstrain ((mu)epsilon). Both strain-related and 17beta-estradiol-related increases in [3H]thymidine incorporation were abolished by the estrogen receptor (ER) modulator ICI 182,780 (10-8 M). Tamoxifen (10(-9) - 10(-8) M) increased [3H]thymidine incorporation in both cell types but had no effect on their response to strain. In TE85 cells, tamoxifen reduced the increase in [3H]thymidine incorporation associated with 17beta-estradiol to that of tamoxifen alone but had no such effect in fHOBs. In TE85 cells, strain increased medium concentrations of insulin-like growth factor (IGF) II but not IGF-I, whereas 17beta-estradiol increased medium concentrations of IGF-I but not IGF-II. Neutralizing monoclonal antibody (MNAb) to IGF-I (3 microg/ml) blocked the effects of 17beta-estradiol and exogenous truncated IGF-I (tIGF-I; 50 ng/ml) but not those of strain or tIGF-II (50 ng/ml). Neutralizing antibody to IGF-II (3 microg/ml) blocked the effects of strain and tIGF-II but not those of 17beta estradiol or tIGF-I. MAb aIR-3 (100 ng/ml) to the IGF-I receptor blocked the effects on [3H]thymidine incorporation of strain, tIGF-II, 17beta-estradiol, and tIGF-I. HOBs and TE85 cells, act similarly to rat primary osteoblasts and ROS 17/2.8 cells in their dose-related proliferative responses to strain and 17beta estradiol, both of which can be blocked by the ER modulator ICI 182,780. In TE85 cells (as in rat primaries and ROS 17/2.8 cells), the response to 17beta estradiol is mediated by IGF-I, and the response to strain is mediated by IGF-II. Human cells differ from rat cells in that tamoxifen does not block their response to strain and reduces the response to 17beta-estradiol in TE85s but not primaries. In both human cell types (unlike rat cells) the effects of strain and IGF-II as well as estradiol and IGF-I can be blocked at the IGF-I receptor. PMID- 11924573 TI - Adjusting bone mass for differences in projected bone area and other confounding variables: an allometric perspective. AB - The traditional method of assessing bone mineral density (BMD; given by bone mineral content [BMC] divided by projected bone area [Ap], BMD = BMC/Ap) has come under strong criticism by various authors. Their criticism being that the projected bone "area" (Ap) will systematically underestimate the skeletal bone "volume" of taller subjects. To reduce the confounding effects of bone size, an alternative ratio has been proposed called bone mineral apparent density [BMAD = BMC/(Ap)3/2]. However, bone size is not the only confounding variable associated with BMC. Others include age, sex, body size, and maturation. To assess the dimensional relationship between BMC and projected bone area, independent of other confounding variables, we proposed and fitted a proportional allometric model to the BMC data of the L2-L4 vertebrae from a previously published study. The projected bone area exponents were greater than unity for both boys (1.43) and girls (1.02), but only the boy's fitted exponent was not different from that predicted by geometric similarity (1.5). Based on these exponents, it is not clear whether bone mass acquisition increases in proportion to the projected bone area (Ap) or an estimate of projected bone volume (Ap)3/2. However, by adopting the proposed methods, the analysis will automatically adjust BMC for differences in projected bone size and other confounding variables for the particular population being studied. Hence, the necessity to speculate as to the theoretical value of the exponent of Ap, although interesting, becomes redundant. PMID- 11924575 TI - Pictorial CME. Spastic quadriplegia due to cervical cord compression. PMID- 11924574 TI - Ciprofloxacin-resistant typhoid with incomplete response to cefotaxime. PMID- 11924576 TI - Theriogenology question of the month. Mineralization of the puppies was suggestive that the bitch would whelp in approximately 11 days. PMID- 11924577 TI - Results of root canal treatment in dogs: 127 cases (1995-2000). AB - OBJECTIVE: To evaluate results of root canal treatment in dogs. DESIGN: Retrospective study. SAMPLE POPULATION: 127 tooth roots in 64 dogs. PROCEDURE: Radiographs obtained before surgery, immediately after surgery, and during follow up examinations after surgery were evaluated by 2 individuals. Treatment was considered successful if the periodontal ligament space was normal and possible preoperative root resorption, if present, had ceased. Treatment was considered to show no evidence of failure if possible preoperative root resorption had ceased but a preexisting periapical lesion had remained the same or only decreased in size and not complete resolved. Treatment was considered to have failed if a periapical lesion or root resorption developed subsequent to endodontic treatment, if a preexisting periapical lesion had increased in size, or if possible preoperative root resorption appeared to continue after endodontic treatment. RESULTS: Follow-up time ranged from 1 to 60 months (mean, 13 months). Treatment was classified as successful for 87 (69%) roots, as showing no evidence of failure for 33 (26%) roots, and as having failed for 7 (6%) roots. The success rate was lower for canine teeth than for maxillary fourth premolar teeth. Roots with a preexisting periapical lucency or preexisting root resorption had lower success rates. The use of intracanal medication and the method and quality of obturation were not associated with outcome. CONCLUSIONS AND CLINICAL RELEVANCE: Results suggest that root canal treatment offers a viable option for salvage of periodontally sound but endodontically diseased teeth in dogs. PMID- 11924578 TI - Three wars that never happened. AB - This article discusses three serious wars that were averted and the three men who averted them. In 1478-79, Pope Sixtus IV's hatred of the Medici culminated in aggressive war against Florence, supported by his powerful ally King Ferrante of Naples. The initial stags of this war were indecisive, but it was about to become much more serious, probably involving all the Italian states and possibly meaning the total destruction of Florence. Lorenzo il Magnifico sailed to Naples, convinced Ferrante this more serious war was against his interests and obtained a generous peace. In 1861, the British Government responded to the boarding of a British ship by a vessel of the American North with a peremptory letter. Albert, Prince Consort, though dying of typhoid fever amended the letter to save Lincoln's face and thus averted war with the North. From 1871 to 1890, Otto von Bismarck worked for a stable peace between the European powers to be attained by arranging meetings of most or all of them to accustom them to solving disputes by negotiation. Two such meetings in Berlin secured 36 years of peace between the powers, despite many disputes, and in particular averted war for possessions in Africa, which could have involved them all. PMID- 11924579 TI - The study of modified calcium hydroxides with surfactants for acid gas removal during incineration. AB - The primary objective of the present work is to use additives to extend the sulfation reaction of the calcium hydroxide (Ca(OH)2) used to control SO2 emission from incineration processes. There are two reasons for adding surfactants (surface-active agent): (1) to provide an appropriate dispersion of the Ca(OH)2', thus preventing particle agglomeration due to humidity; (2) to alter the sulfation reaction environment by adsorbing heterogeneous materials on the surface of the Ca(OH)2 to extend the adsorption equilibrium. A dry scrubber integrated with a fabric filter was employed to study the effect of surfactants on the removal efficiency of acid gas in the flue gas with Ca(OH)2 as the sorbent. The operating parameters evaluated include: (1) the different surfactants (calcium lignosulfonate, sodium lignosulfonate, alkyl naphthalene sodium sulfonate and beta-naphthalene sodium sulfonate condensates) and (2) the composition of acid gas (i.e. sulfur dioxide (SO2), nitrogen oxide (NO) and hydrogen chloride (HCl)). The results show that modified Ca(OH)2 with surfactants could effectively decrease the emission of acid gas during incineration. Different additives had individual absorption efficiencies on different acid gases. On the whole, sodium lignosulfonate and beta-naphthalene sodium sulfonate condensates had better sorption capacity for SO2 and NO, but not for HCl. In addition, when SO2 coexisted with NO and HCl, the concentration of NO and HCl will result in decrease or increase of the removal efficiency of SO2. PMID- 11924580 TI - Pig manure as a co-composting material for biodegradation of PAH-contaminated soil. AB - Pig manure at three different ratios of 12.5%, 25% and 50% (w/w dry weight basis) was amended with a soil spiked with 100 mg kg(-1) each of three polycyclic aromatic hydrocarbons (phenanthrene, anthracene, and pyrene) to investigate its effect on the biodegradation of these PAHs in a bench-scale composting system. An increase in pig manure amendment was effective in enhancing the amounts of soluble organic carbon, ammoniacal nitrogen, and soluble phosphorous in the composting mass. It could also increase the populations of total thermophilic and mesophilic bacteria as well as PAH-degrading bacteria, but this pattern was restricted only to the early stage of the composting process. High amounts of pig manure in the composting mass reduced the seed germination or root growth of cress seeds, but the composting process was effective in reducing the phytotoxic effects of the compost. Amendment of pig manure was beneficial to PAH removal during composting treatment and maximum removal rate at the end of composting accounted for 90% of the initial concentrations of PAHs. A pig manure application rate of 25% showed the most efficient removal of 3-ringed PAHs (phenanthrene and anthracene), while no significant difference in pyrene removal for those receiving 25 or 50% pig manure amendment. Taking into consideration the effects of pig manure on seed germination and available nutrients in the composting mass, this study suggested that a pig manure amendment of 25%, i.e., 3:1 ratio of contaminated soil: pig manure, is recommended to co-compost with PAH-contaminated soil. PMID- 11924581 TI - Denitrification of high NO3(-)-N containing wastewater using elemental sulfur; nitrogen loading rate and N2O production. AB - The effects of both concentration and loading rates of nitrate on denitrification efficiency and N2O production during the autotrophic denitrification with elemental sulfur were evaluated. Experiments were conducted with three continuously fed sulfur packed bed reactors (SPBRs) with different influent nitrate concentrations. The loading rate at which nitrate removal efficiency was greater than 95% decreased from 2.46 to 1.64 kg NO3(-)-N m(-3) day(-1) as the influent NO3(-)-N concentration increased from 175 to 700 mg 1(-1). The nitrate removal rate per unit volume of sulfur layer can be expressed as alpha (ALR)n, where ALR is the applied loading rate. The range of alpha and n values were 0.72 0.83 and 0.91-0.94, respectively. The maximum nitrate removal rate was estimated as 3.9 to 4.5 kg NO3(-)-N m(-3) day(-1) depending on the influent NO3(-)-N concentration. The loading rate for complete denitrification without N2O production was 1.37 kg NO3(-)-N m(-3) day(-1) at the influent NO3(-)-N concentration of 175 mg 1(-1). N2O production became significant when the influent nitrate concentration increased while the loading rate remained the same. The batch test indicated that the amount of N2O production per unit mass of nitrogen was greater with nitrite than with nitrate showing that N2O production was directly related to the concentration of nitrite and nitrate. PMID- 11924582 TI - Gamma ray detection using sol-gel glass doped with lanthanide ions. AB - In this work we discuss the synthesis, characterization and the use of rare earth doped silica glass prepared by the sol-gel technique as a gamma radiation detector. We obtain a glass material doped with rare earth ions in a high local symmetry, in contrast to the low symmetry found in glasses produced by conventional melting methods and technologies. The behavior of the luminescence spectra of the excited states of rare earth indicates a strong dependence with gamma radiation doses, where the magnetic dipole transition 5D0-->7F1 of the Eu3+ ions presents an huge enhancement of 900% for irradiation doses up to 400 Gy. PMID- 11924583 TI - [A study of job and smoking habits in Japanese hospital nurses]. PMID- 11924584 TI - Apparent resistance of Parascaris equorum to macrocylic lactones. PMID- 11924585 TI - Primary central chondrosarcoma. PMID- 11924586 TI - Oxidation of orthopaedic UHMWPE. AB - Retrieved EtO sterilised acetabular cups usually show much less degradation than gamma-ray sterilised cups. Some of our retrieved EtO sterilised cups did, however, reveal unexpected bulk oxidation. It was observed that this oxidation was always accompanied by whitening of the material. This whitening was found to be due to a break-up of the compression moulded material into its original particles. It was noticed that there was no oxidation in all parts, where the break-up and whitening appeared. The oxidation did, however, occur exclusively in the parts where there was a badly consolidated material. Upon examining shelf aged, unsterilised samples, it was found that the degradation was also present here. This shows that the observed phenomenon is not due to the service in vivo and that it must originate from the processing step. Just as for the retrieved samples, the shelf aged cups only showed oxidation in the bulk and not at the surface. It was concluded that the material used for the cups had been badly fused together during the compression moulding and that the machining had created a bad stress situation in the cups leading to a break-up of the particles. The mechanism that initiates the oxidation is not known, but it is believed that the distribution depends on how the internal stresses have acted to break up the structure. In the areas where the particles have been separated, there is probably a higher availability of oxygen than what is normally observed in UHMWPE. PMID- 11924587 TI - Immobilization of a nonsteroidal antiinflammatory drug onto commercial segmented polyurethane surface to improve haemocompatibility properties. AB - A method has been developed in which a layer of p-aminosalicylic acid (4-amino-2 hydroxybenzoic acid) (PAS), a water soluble pharmaceutical compound of the nonsteroidal anti-inflammatory drug (NSAID) class with antiaggregant platelet activity, is covalently immobilized onto a segmented polyurethane, Biospan (SPU) surface. Thus, SPU surfaces were modified by grafting of hexamethylenediisocyanate. and the free isocyanate remaining on the SPU surface were then coupled through a condensation reaction to amine groups of p aminosalicylic acid. The bonding of PAS from aqueous solution onto SPU surface was studied by ATR-FTIR. UV and fluorescence spectroscopy. Plateau levels of coupled PAS were reached within 1.2 microg/cm2 using PAS solution concentrations of 1mg/ ml. The surface wettability of the polymeric films measured by contact angle indicate that the introduction of the PAS turns the surface more hydrophilic (theta(water) = 43.1 +/- 2.1) relatively to the original SPU films (theta(water) = 70.3 +/- 1.9). The in vitro albumin (BSA) adsorption shows that the PAS-SPU films adsorb more BSA (250/microgmm2) than the original SPU (112 microg mm2). Thrombogenicity was assessed by measuring the thrombus formation and platelet adhesion of the SPU containing PAS relatively to nonmodified SPU surfaces. The polymeric surfaces with immobilized PAS had better nonthrombogenic characteristics as indicated by the low platelet adhesion, high adsorption of albumin relatively to fibrinogen and low thrombus formation, making them potentially good candidates for biomedical applications. PMID- 11924588 TI - Degradable polyphosphazene/poly(alpha-hydroxyester) blends: degradation studies. AB - Biomaterials based on the polymers of lactic acid and glycolic acid and their copolymers are used or studied extensively as implantable devices for drug delivery, tissue engineering and other biomedical applications. Although these polymers have shown good biocompatibility, concerns have been raised regarding their acidic degradation products, which have important implications for long term implantable systems. Therefore, we have designed a novel biodegradable polyphosphazene/poly(alpha-hydroxyester) blend whose degradation products are less acidic than those of the poly(alpha-hydroxyester) alone. In this study, the degradation characteristics of a blend of poly(lactide-co-glycolide) (50:50 PLAGA) and poly[(50% ethyl glycinato)(50% p-methylphenoxy) phosphazene] (PPHOS EG50) were qualitatively and quantitatively determined with comparisons made to the parent polymers. Circular matrices (14mm diameter) of the PLAGA, PPHOS-EG50 and PLAGA-PPHOS-EG50 blend were degraded in non-buffered solutions (pH 7.4). The degraded polymers were characterized for percentage mass loss and molecular weight and the degradation medium was characterized for acid released in non buffered solutions. The amounts of neutralizing base necessary to bring about neutral pH were measured for each polymer or polymer blend during degradation. The poly(phosphazene)/poly(lactide-co-glycolide) blend required significantly less neutralizing base in order to bring about neutral solution pH during the degradation period studied. The results indicated that the blend degraded at a rate intermediate to that of the parent polymers and that the degradation products of the polyphosphazene neutralized the acidic degradation products of PLAGA. Thus, results from these in vitro degradation studies suggest that the PLAGA-PPHOS-EG50 blend may provide a viable improvement to biomaterials based on acid-releasing organic polymers. PMID- 11924589 TI - Disodium chlodronate prevents bone resorption in experimental periodontitis in rats. AB - BACKGROUND: Periodontitis is a chronic disease characterized by alveolar bone loss and inflammatory changes. We studied the effect of disodium chlodronate (CD), a bisphosphonate used in metastatic and metabolic bone disease as a bone resorbing drug, in an experimental periodontitis model (EPD) focusing on anti resorptive and anti-inflammatory parameters. METHODS: A nylon thread ligature was placed around the left maxillary molars of 72 male Wistar rats who were sacrificed after 7 or 11 days. Groups were treated daily with CD (1, 5, or 25 mg/kg/sc) starting at day 0 until day 7 (prophylactic CD) or from day 5 until day 11 (curative CD) after periodontitis induction. Non-treated group (NT) consisted of rats subjected to periodontitis that received no pharmacological treatment. Alveolar bone loss (ABL) was measured as the distance between the cuspid tips and the alveolar bone. The right jaw was used as control. The hemiarcades were processed for histopathologic analysis. RESULTS: In NT group there was significant ABL, severe mononuclear cells influx, and increase in osteoclast numbers. Prophylactic CD treatment decreased the ABL 25.8%, 61.6%, and 75.5% as compared to NT for the 1, 5, and 25 mg/kg CD doses, respectively. Curative CD treatment decreased the ABL 20%, 62%, and 69% as compared to NT for the 1, 5 and 25 mg/kg CD doses, respectively. Both prophylactic and curative CD decreased histological changes, as compared to NT rats (P <0.01). CONCLUSIONS: CD has both bone sparing and anti-inflammatory activity in EPD in rats when administered as a pretreatment or in an ongoing process. The possibility of using CD as an alternative treatment in human periodontitis should be considered. PMID- 11924590 TI - Efficacy of chlorhexidine mouthrinses with and without alcohol: a clinical study. AB - BACKGROUND: Plaque control is the main method for preventing periodontal diseases. Chlorhexidine digluconate mouthrinse is widely recognized as helping to maintain plaque control. Most of these mouthrinses contain alcohol, making them impractical for many patients, including those with oral mucosal hypersensitivity. Mouthrinses without alcohol might cause fewer side effects, but also be less efficient. In this study, we evaluated the efficacy of a 0.12% chlorhexidine mouthrinse without alcohol against one with 11% ethanol and a placebo. METHODS: This a double-blind, parallel group study with 96 patients who tested 3 mouthrinses containing 1) chlorhexidine digluconate 0.12% sodium fluoride 0.05%, and ethanol 11% (group 1; CHX-A); 2) the same solution without alcohol (group 2; CHX-NA); and a placebo (group 3; P). Plaque and bleeding indexes were recorded in all patients prior to treatment and at 14 and 28 days. RESULTS: There were significant differences in plaque, gingivitis, and papilla bleeding indexes in both chlorhexidine rinses compared to placebo, but no differences between the 2 CHX products. CONCLUSIONS: In this study, the alcohol free rinse was as effective as one containing alcohol in controlling plaque and reducing gingival inflammation. Therefore, it would seem that its use can be recommended in all patients, but especially in patients for whom the use of alcohol is contraindicated. PMID- 11924591 TI - Rational design of a polymer specific for microcystin-LR using a computational approach. AB - A computational approach for the design of a molecularly imprinted polymer (MIP) specific for Cyanobacterial toxin microcystin-LR is presented. By using molecular modeling software, a virtual library of functional monomers was designed and screened against the target toxin, employed as a template. The monomers giving the highest binding energy were selected and used in a simulated annealing (molecular dynamics) process to investigate their interaction with the template. The stoichiometric ratio observed from the simulated annealing study was used in MIP preparation for microcystin-LR. The monomers were copolymerized with a cross linker in the presence of the template. A control (blank) polymer was prepared under the same conditions but in the absence of template. A competitive assay with microcystin-horseradish peroxidase conjugate was optimized and used to evaluate the affinity and cross-reactivity of the polymer. The performance of the artificial receptor was compared to the performance of monoclonal and polyclonal antibodies raised against the toxin. The results indicate that imprinted polymer has affinity and sensitivity comparable to those of polyclonal antibodies (the detection limit for microcystin-LR using the MIP-based assay was found to be 0.1 microg L-1), while superior chemical and thermal stabilities were obtained. Moreover, cross-reactivity to other toxin analogues was very low for the imprinted polymer, in contrast to the results achieved for antibodies. It is anticipated that the polymer designed could be used in assays, sensors, and solid phase extraction. PMID- 11924592 TI - Detection of hydrogen peroxide at mesoporous platinum microelectrodes. AB - Mesoporous (H(I)-ePt) platinum microelectrodes electrodeposited from the hexagonal (H(I)) lyotropic liquid crystalline phase are shown to be excellent amperometric sensors for the detection of hydrogen peroxide over a wide range of concentrations. Good reproducibility, high precision, and accuracy of measurements are demonstrated. Mesoporous microelectrodes retain the high rates of mass transport typical of conventional microelectrodes, and their high real surface area greatly enhances their catalytic activity. This unique combination of properties overcomes the limitations of previous amperometric hydrogen peroxide sensors and yields outstanding qualitative and quantitative results. PMID- 11924593 TI - An in-depth analysis of electric cell-substrate impedance sensing to study the attachment and spreading of mammalian cells. AB - The attachment and spreading of fibroblast cells on a gold surface coated with fibronectin or ovalbumin were studied by a modified electric cell-substrate impedance sensor. In this system, cells were cultured in a well, equipped with a detecting gold electrode (surface area of 0.057 mm2) and a gold counter electrode (18 mm2). Based on a comprehensive theoretical framework, the impedance of the electrode-electrolyte interface and a cell layer was precisely obtained for frequencies ranging from 1 to 10 kHz. Surface concentrations of the protein adsorbed on the gold surface were determined by a surface plasmon resonance biosensor. The resistance change of the electrode-electrolyte interface at 4 kHz increased linearly with the number of fibroblast cells attached on the detecting electrode. The slope of the linear relationship appeared to depend on the type of coating protein. As the surface area occupied by the cells was also proportional to the cell number, the resistance change was in turn proportional to the area covered by the cells. PMID- 11924594 TI - Thermal unfolding of proteins studied by coupled reversed-phase HPLC-electrospray ionization mass spectrometry techniques based on isotope exchange effects. AB - In this study, a deuterium exchange procedure has been employed to evaluate the thermal stability of globular proteins under conditions that replicate their interactive behavior in reversed-phase high performance chromatographic (RP-HPLC) systems. In particular, this investigation has permitted the conformational stability of two proteins, hen egg white lysozyme (HEWL) and horse heart myoglobin (HMYO) to be examined under different temperature and low-pH solvent regimes. The results confirm that this experimental approach provides an efficient strategy to explore fundamental conformational features of polypeptides or proteins in their folded and partial unfolded states under these interactive conditions. In particular, this analytical procedure permits insight to be readily gained into the processes that occur when polypeptides and globular proteins interact with lipophilic liquid/ solid interfaces in the presence of water-organic solvent mixtures at different temperatures. PMID- 11924595 TI - The spectrum of subdivided left atrium: diagnosis, treatment and outcome in eight patients. AB - From January 1996 to April 2001, we treated eight patients with subdivided left atrium. Their ages at diagnosis ranged from newborn to 6.4 years. Prominent clinical symptoms were failure to thrive, respiratory symptoms, demand for oxygen, and congestive cardiac failure. Diagnosis was made by transthoracic echocardiography in all cases. Cardiac catheterization was necessary only in those patients who had associated cardiac anomalies, or suspected signs of pulmonary hypertension. In 7 patients, surgery was performed immediately after diagnosis, but one preterm infant died before operation due to neonatal sepsis and respiratory distress syndrome. In the postoperative period, one patient developed a severe capillary leak syndrome, and died due to irreversible congestive cardiac failure. The other 6 patients have all been followed up, with good results in the short- and intermediate-term at a mean of 34.3+/-20.2 months. The infants were thriving, had a reduction in the frequency of infections of the respiratory tract, no significant arrhythmias, and showed early recovery from pulmonary hypertension and right ventricular hypertrophy. PMID- 11924596 TI - Cardiac diverticulum and omphalocele: Cantrell's pentalogy or syndrome. AB - Omphaloceles and left ventricular diverticulums are rare disorders. Although either is known to occur on its own, the combination is highly suggestive of the so-called pentalogy of Cantrell. This syndrome is a combination of deformities involving midline structures, with exteriorisation of the heart, or 'ectopia cordis', as the most severe malformation. A cause has yet to be identified, though genes located on the X-chromosome may be involved. We discuss a neonate who presented with an omphalocele and a palpable diverticulum of the left ventricle. An omphalocele, especially when above the umbilicus, is an indication for further investigation for deformities as seen in the spectrum of Cantrell's pentalogy, especially cardiac malformations and anterior diaphragmatic herniation. A left ventricular diverticulum is usually associated with Cantrell's syndrome. When found, it is usually accompanied by other intracardiac malformations, so that again further examination is indicated. In our patient, there was an atrial septal defect within the oval fossa, along with a ventricular septal defect and unobstructed albeit anomalous venous pulmonary return to the left atrium. Early surgical intervention seems to be indicated, as spontaneous rupture, arrhythmias, and thromobogenicity of the ventricular diverticulum have all been reported. PMID- 11924597 TI - Hplc detection of patulin in apple juice with GC/MS confirmation of patulin identity. AB - The official patulin LC procedure was further examined (AOAC 995.10). Juice or juice concentrate was extracted with ethyl acetate and cleaned up with sodium carbonate. Patulin in the dried extract was determined by reversed-phase LC with UV detection (280 nm) in 1% THF aqueous solution after evaporation of the ethyl acetate. An end-capped C18 column was required to separate patulin from hydroxymethylfurfural. Patulin was detected in approximately half of the >1000 extracts examined. Only ca 10% of the extracts contained patulin at levels greater than 50 microg/L (50 ppb). Some presumptive findings were confirmed by capillary gas chromatography/mass spectrometry as the trimethyl silyl derivative using electron ionization or as underivatized patulin using negative ion chemical ionization. Trifluoropropylmethyl polysiloxane capillary columns provided superior gas chromatography of underivatized patulin compared to phenyl/methyl polysiloxane and methyl polysiloxane columns. PMID- 11924598 TI - Risk assessment of deoxynivalenol in food: concentration limits, exposure and effects. AB - The mycotoxin, deoxynivalenol (DON), is produced world-wide by the Fusarium genus in different cereal crops. We derived a provisional TDI of 1.1 microg/kg body weight (bw) and proposed a concentration limit of 129 microg DON/kg wheat based on this TDI and a high wheat consumption of children. In the period September 1998-January 2000, the average DON concentration in wheat was 446 microg/kg (n = 219) in The Netherlands. During this period, the dietary intake of DON exceeded the provisional TDI, especially in children. Eighty percent of the one-year-olds showed a DON intake above the provisional TDI and 20% of these children exceeded twice the provisional TDI. Our probabilistic effect assessment shows that at these exposure levels, health effects may occur. Suppressive effects on body weights and relative liver weight were estimated at 2.2 and 2.7%. However, the large confidence intervals around these estimates indicated that the magnitudes of these effects are uncertain. PMID- 11924599 TI - Morphological characterisation of three isolates of Heterorhabditis Poinar, 1976 from the 'Irish group' (Nematoda: Rhabditida: Heterorhabditidae) and additional evidence supporting their recognition as a distinct species, H.downesi n.sp. AB - The morphological variation of three representative isolates of the 'Irish group' of Heterorhabditis was examined. First generation hermaphrodites were characterised by having a blunt and mucronate tail. Females (second generation) and third-stage infective juveniles were also distinguished by the morphology of the tail and the presence of a refractile projection in the tail tip. Males were characterised by the position of the excretory pore and by the value of ratio SW. These morphological features do not fit the description of currently recognised Heterorhabditis species, and provide additional evidence in support for the consideration of the Irish group as a new species. A description of this species, as H. downesi n. sp., is provided. PMID- 11924600 TI - [Colonoscopy or sigmoidoscopy: risk of isolated right colon lesions]. AB - BACKGROUND: Colorectal cancer is an important cause of death in western countries. Screening methods are based on flexible sigmoidoscopy, a cheap, effective, and less painful procedure, but some important lesions on the right colon can be missed. AIM: Evaluate how many important lesions would be missed if colonoscopy indicated only for patients with distal lesions identified during flexible sigmoidoscopy. MATERIAL AND METHODS: All consecutive colonoscopy performed in the Endoscopy Unit of the Gastroenterology Division of Federal University of Sao Paulo, SP, Brazil, with polyps and cancer suspicious lesions were reviewed. Patients with a second procedure, inflammatory bowel disease or colonic surgery were excluded. RESULTS: We reviewed 101 patients with 38 possible cancer lesions and 70 polyps. The mean age was 62 years (SD 13.7 years) and 45 were male. Forty five lesions were in the right colon, namely 23 adenomas, 10 adenocarcinomas, 1 lymphoma and 11 benign lesion. Among such lesions, 28 were isolated in the right colon (16 adenomas and 7 adenocarcinomas). CONCLUSION: In our study 23 patients (22.77%) had adenomas or adenocarcinomas isolated in the right colon, that would be missed if flexible sigmoidoscopy was used alone. PMID- 11924602 TI - Simultaneous flow cytometric analysis of phagocytosis and oxidative burst activity in equine leukocytes. AB - This paper describes a method for simultaneously measuring phagocytosis and oxidative burst activity in equine peripheral blood leukocytes by flow cytometry. Opsonized propidium iodide-labelled Staphylococcus aureus (PI-Sa) was used to measure the uptake of bacteria by equine phacocytes and the oxidative burst activity by oxidation of dihydrorhodamine 123. The requirements to achieve optimal activity of phagocytosis and oxidative burst are described. The advantage of the simultaneous technique is that it provides both independent and comparative values for phagocytosis and the oxidative burst, for the detection of impaired mechanisms of microbial destruction. Furthermore, the technique allows evaluation of opsonization activity in this context. PMID- 11924603 TI - Pathogenicity of a Hong Kong-origin H5N1 highly pathogenic avian influenza virus for emus, geese, ducks, and pigeons. AB - The H5N1 type A influenza viruses that emerged in Hong Kong in 1997 are a unique lineage of type A influenza viruses with the capacity to transmit directly from chickens to humans and produce significant disease and mortality in both of these hosts. The objective of this study was to ascertain the susceptibility of emus (Dramaius novaehollandiae), domestic geese (Anser anser domesticus), domestic ducks (Anas platyrhynchos), and pigeons (Columba livia) to intranasal (i.n.) inoculation with the A/chicken/Hong Kong/220/97 (H5N1) highly pathogenic avian influenza virus. No mortality occurred within 10 days postinoculation (DPI) in the four species investigated, and clinical disease, evident as neurologic dysfunction, was observed exclusively in emus and geese. Grossly, pancreatic mottling and splenomegaly were identified in these two species. In addition, the geese had cerebral malacia and thymic and bursal atrophy. Histologically, both the emus and geese developed pancreatitis, meningoencephalitis, and mild myocarditis. Influenza viral antigen was demonstrated in areas with histologic lesions up to 10 DPI in the geese. Virus was reisolated from oropharyngeal and cloacal swabs and from the lung, brain, and kidney of the emus and geese. Moderate splenomegaly was observed grossly in the ducks. Viral infection of the ducks was pneumotropic, as evidenced by mild inflammatory lesions in the respiratory tract and virus reisolation from oropharyngeal swabs and from a lung. Pigeons were resistant to HK/220 infection, lacking gross and histologic lesions, viral antigen, and reisolation of virus. These results imply that emus and geese are susceptible to i.n. inoculation with the HK/220 virus, whereas ducks and pigeons are more resistant. These latter two species probably played a minimal epidemiologic role in the perpetuation of the H5N1 Hong Kong-origin influenza viruses. PMID- 11924601 TI - Present concepts on the inflammatory modulators with special reference to cytokines. AB - The pro- and anti-inflammatory cytokines create a network of interactions between cells that lead to both stimulatory and inhibitory responses that maintain an effective homeostatic regulation. The anti-inflammatory cytokines are a family of peptides that modulate the pro-inflammatory cytokine response. Cytokines act in concert with non-cytokine mediators, such as prostaglandin E2, glucocorticosteroids, lipocortins, and catecholamines. This review highlights new developments in our understanding of the pathophysiology of inflammation and gives an example of a more recent approach to the modulation of acute systemic inflammatory disorders; activation of beta2-adrenergic receptors on macrophages. In this respect the potent beta2-adrenergic agonist clenbuterol seems of therapeutic interest. PMID- 11924604 TI - Experimental reproduction of malabsorption syndrome with different combinations of reovirus, Escherichia coli, and treated homogenates obtained from broilers. AB - Attempts to reproduce malabsorption syndrome (MAS) by oral inoculation with several different combinations including intestinal homogenate, reovirus, and hemolytic Escherichia coli obtained from MAS-affected chickens and intestinal homogenate from healthy chickens (healthy homogenate) were performed in 1-day-old specific-pathogen-free (SPF) broilers. The MAS homogenate, serving as a positive control, induced weight gain depression and intestinal lesions such as cystic crypts of Lieberkuhn, villus atrophy, and lymphoid and/or granulocytic infiltration. The healthy homogenate, the formalin-treated MAS homogenate, the formalin-treated healthy homogenate, and phosphate-buffered saline caused neither weight gain depression nor intestinal lesions. We were able to reproduce both weight gain depression and intestinal lesions by inoculation of reovirus either combined with the formalin-treated MAS homogenate or combined with healthy homogenate. Surprisingly, when hemolytic E. coli was added to the combination of reovirus with formalin-treated MAS homogenate, this did not cause weight gain depression although this combination caused the described intestinal lesions. Identical results were obtained with the combination of formalin-treated MAS homogenate with hemolytic E coli or the combination of reovirus with hemolytic E. coli. The intestinal lesions were more severe and developed faster by combinations including reovirus and formalin-treated MAS homogenate. This study indicates that a combination of enteropathogenic reovirus with other agents or substances that are present in an intestinal homogenate from MAS-affected and healthy chickens can induce MAS in SPF broilers. Escherichia coli is not essential for induction of weight gain depression but can play a role in development of intestinal lesions. Furthermore, intestinal lesions alone will not always result in weight gain depression. PMID- 11924605 TI - Axonal transport of neurofilaments in normal and disease states. AB - Neurofilaments are among the most abundant organelles in neurones. They are synthesised in cell bodies and then transported into and through axons by a process termed 'slow axonal transport' at a rate that is distinct from that driven by conventional fast motors. Several recent studies have now demonstrated that this slow rate of transport is actually the consequence of conventional fast rates of movement that are interrupted by extended pausing. At any one time, most neurofilaments are thus stationary. Accumulations of neurofilaments are a pathological feature of several human neurodegenerative diseases suggesting that neurofilament transport is disrupted in disease states. Here, we review recent advances in our understanding of neurofilament transport in both normal and disease states. Increasing evidence suggests that phosphorylation of neurofilaments is a mechanism for regulating their transport properties, possibly by promoting their detachment from the motor(s). In some neurodegenerative diseases, signal transduction mechanisms involving neurofilament kinases and phosphatases may be perturbed leading to disruption of transport. PMID- 11924606 TI - Aged care, ethics and public policy. PMID- 11924607 TI - On being cared for by a nurse. AB - The hospitalised patient's experiences of nursing care, the nature and the value of the patient's and nurse's relationships are relatively unexplored territory in terms of Australian research. This is despite the importance and centrality of caring to the practice of nursing and the very significant contributions that overseas researchers have made to understanding the phenomenon of care in nursing practice. Given the social, economic and cultural differences between Australia and other countries, a study which examines Australian perspectives of the nursing caring experience is long overdue. That caring makes a difference to the patient's sense of well being should come as no surprise to those of us who care. In this paper the writer aims to demonstrate how that difference comes about. PMID- 11924608 TI - Rehabilitation nursing in Australia: a valid and valued specialty. AB - While rehabilitation nursing has been largely overlooked in Australia, it is becoming more visible. Its invisible nature is highlighted when a comparison to acute care nursing is made. However, through the activities of the rehabilitation nurses' professional nursing organisation and recent research there is a growing awareness of the nature of rehabilitation. That rehabilitation does not fit the current classifications for nursing specialties is also highlighted as a problem. This paper aims to expose rehabilitation nursing as a valid and valued specialty. PMID- 11924609 TI - The value of asthma camps for young people in Victoria, Australia. AB - PURPOSE: This paper investigates the impact of the Asthma Foundation of Victoria's educational camp program on children's knowledge of asthma and its management, their feelings about asthma, and their attitudes toward physical and social activities. Parents' observations of changes in their child's behaviour and attitudes are also reported. DESIGN AND METHODS: This research was descriptive and applied. It used questionnaires atfour stages (directly pre- and post-camp, three-four months and ten-15 months post-camp) of an asthma education camp program to assess child asthma knowledge levels. At three months post-camp, parental observations of children's attitudes and behaviours were assessed using a questionnaire. Children's feelings toward asthma were also assessed using a questionnaire pre- and post-camp. RESULTS: The children surveyed displayed a better knowledge of asthma and how to manage their condition immediately after the camp. This knowledge tended to return to pre-camp levels after ten months. The children also reported less anxiety and fear about their illness, a greater sense of wellbeing, and more confidence in participating in a whole range of physical and social activities. Many parents also noted positive changes in their children in terms of activities and asthma management at three months post-camp. CLINICAL IMPLICATIONS: Although there were limitations to sustaining knowledge gained in the asthma camping program, the camping experience provided a benefit for children in terms of promoting their mental and social wellbeing. When readers consider modernising asthma education (e.g. shorter camps, education in everyday social settings such as schools), they need to consider retaining the key ingredients of the more traditional camping program that supports good asthma management, wellbeing and social participation. PMID- 11924610 TI - Priorities for paediatric cancer nursing research in Western Australia: a Delphi study. AB - A Delphi study was undertaken to identify paediatric cancer nursing research priorities at the sole paediatric tertiary referral centre in Western Australia. Thirty one responses were received from the first Delphi round and 22 from the second round. The top 10 priorities identified were different from those identified in prior North American Delphi studies. The top ranked research topic was 'To effective methods of communication in a multi-disciplinary identify health care team'. The findings will provide a research agenda for cancer nursing at the study setting and may stimulate discussion and re-assessment of research priorities in other paediatric cancer settings. PMID- 11924611 TI - How do early parenting centres support women with an infant who has a sleep problem? AB - Some women who have an infant experiencing sleep problems arc so sleep deprived themselves that admission to an early parenting centre is needed to help them better manage the situation. This paper reports on a qualitative sttudy that used focus group methods to interview families who were admitted to an early parenting centre for persistent infant sleep problems. Results showed that parents needed a variety of strategies such as instructional, emotional and physical supports to develop confidence to manage infant's sleep disturbance at home. PMID- 11924612 TI - Retaining a viable workforce: a critical challenge for nursing. AB - Nursing is facing a crisis nationally and internationall, with Australia, the United States, New Zealand, Canada, the United Kingdom and Western Europe experiencing critical shortages of nurses. Problems with recruitment, retention and an ageing workforce means that attempts to ensure a viable nursing workforce must be placed at the top of the professional agenda. Strategies currently used to manage the crisis, such as overseas recruitment, are not sustainable and are ethically dubious. The demographic timebomb is ticking and up to half the current nursing workforce will reach retirement age by 2020. It is vital that there are adequate numbers of skilled and qualified nurses to take their places. Nursing and nurses are facing unprecedented challenges and pressures in the workplace. Job satisfaction is threatened as nurses are pressured to do more with less, Nursing productivity has increased phenomenally over the past ten years in response to increased demands and decreasing numbers of staff. The nursing workplace has disturbingly high levels of occupational violence, and many nurses operate within a culture of blame and scapegoating. There is evidence that organizational change is imposed upon nurses with little or no consultation and the literature reveals that this has a direct and negative effect on job satisfaction and on retention of nurses. This paper explores some of the critical issues that nursing must confront to be successful in establishing and maintaining a vigorous, dynamic and viable workforce. PMID- 11924613 TI - Enhancing access to and retention in the Bachelor of Nursing: a successful alternative. AB - This paper describes an innovative program to enhance access to the Bachelor of Nursing (BN) in theform of a nursing entrance test (NET). Discussion will focus on the NET as the meansfor enhancing access and retention. The results of a recent evaluation of the NET are discussed and used to demonstrate that the NET is an effective tool for the provision of access and retention in the Bachelor of Nursing for students who do not meet standard entry criteria. PMID- 11924614 TI - Turning tapes into text: issues surrounding the transcription of interviews. AB - Transcription of interview data is a common practice in qualitative health research. However, there has been little discussion of the techniques of transcription and the issues inherent in the use of transcription as a strategy for managing qualitative data in nursing publications. The process of transcription may disclose or obscure certain information. Researchers need to question practices of transcription that have been taken for granted and make transparent the processes used to preserve the integrity of data. This paperfirst examines research reported in nursing and allied health journals employing interviews for data collection and the attention given to the transcription phase. It then deals with issues of concern regarding the transcription of interviews, and offers suggestions for promoting validity. PMID- 11924615 TI - Overcoming obstacles in becoming a reflective nurse and person. AB - As a practical approach to overcoming obstacles in being a reflective nurse and person, this article confirms the personhood of nurses, and describes some types and methods of reflection, and their uses at work and in everyday home life. Practical and theoretical obstacles are identified, and strategies are offered for overcoming them. PMID- 11924616 TI - An action research approach to the development of a clinical pathway for women requiring Caesarean sections. AB - An action research framework was utilised to introduce a clinical pathway for women undergoing Caesarean section in the Maternity Unit at Westmead Hospital in 1998-1999. This style of research allowed the development of the clinical pathwayfor women having a Caesarean section to be progressively modified to adapt it to suit the needs of the multidisciplinary team and the clinical environment. The clinical pathway itself became a document that facilitated the education of the women, improving their preparation for discharge, aided the orientation and skill development of new staff and improved the continuity of care by enhancing the multidisciplinary approach to the women's care. It also identified the need for continuing staff education throughout the implementation and evaluation phases. PMID- 11924617 TI - Trajectory, transition and vulnerability in adult medical-surgical patients: a framework for understanding in-hospital convalescence. AB - Relatively recent changes in health care have seen shorter hospital stays and a considerable increase in day-only admissions, anticipated to involve 60% of all patients admitted for surgery and/or investigations within the next couple of years. Surprisingly little is known about the experiences of the approximately 40% of patients who will begin to convalesce in hospital following illness, trauma or surgery. Findings from a qualitative study, briefly reported here, identified a sense of vulnerability in transition as a significant discomfort. By examining assumptions underlying the care of patients recovering in hospital, the nexus between system-determined care trajectories transition, and patient vulnerability can be identified and problems suitably addressed. PMID- 11924618 TI - Restructuring of aged care facilities: easing the burden on public hospitals. PMID- 11924619 TI - Which patient should be resuscitated first? AB - The core principles of deontology are justice and beneficence. The principle of justice and egalitarianism guide health care professionals to treat every client with fairness and equity regardless the prognosis of illness, social and economic status of clients, the social and financial consequences impose on others. Whereas, the principle of beneficence and utilitarianism direct health care professionals to make an ethical decision to provide the maximum benefit and to minimise harm to the greatest number of people involved. When distributing limited health care resources, there could be conflicts between the principle of justice and beneficence or between egalitarians and utilitarians. At times health care professionals have to make a painful decision which only complies with certain ethical principles, but does not adhere to others. PMID- 11924620 TI - Identifying barriers to the application of evidence based practice in mental health nursing. AB - Although contemporary nursing literature urges clinicians to embrace the principles of evidence-based practice and utilise research to guide professional activity, this challenge has frequently gone unheeded by mental health nurses. Indeed the reality of merging research and practice is a complex process impeded by significant barriers. Examination of these barriers provides credible reasonfor mental health nurses' slow progress toward evidence-based nursing practice to date. However, exploration of these barriers also leads to the identification of strategies to overcome them, inevitably rendering the current situation untenable. PMID- 11924621 TI - Registered male: a discussion on men in the nursing profession. AB - The aim of this discussion paper is to alert the profession to issues surrounding men in nursing. There is a historical connection between men and the nursing profession; men having served as nurses long before Florence Nightingale. Literature suggests that women and men enter the nursing profession for the same reasons, however the paths which lead men to nursing differ. Feminist sociologists express alarm at the disproportionate number of males in managerial nursing positions and hold that this may have negative consequences for the profession. Male and female patients have a definite preference in regards to the gender of their nurse. Issues surrounding this preference will be explored. PMID- 11924622 TI - The impact of lifespan development and culture on the health of older migrant women. AB - Australia has a long history of accepting migrants from all over the world. As a consequence, one of the challenges in nursing is to provide culturally relevant and culturally safe care to people from varying ethnic backgrounds. One specific issue that nurses need to be aware of is the significance of the impact on migrant women in the "sandwich generation" when providing care their elderly relatives. Nurses can use lifes-pan developmental theory to effectively provide care for migrant groups, by being aware of the developmental level of th e person and its impact. PMID- 11924623 TI - A family's perception of a public hospital. AB - This article tells of one family's recent perceptions of major public hospital. Their story is a personal one hat revolves around the illness of a family member. Visiting their loved one every day meant they gained heightened awareness of the day-to-day running of the hospital. As the patient had to stay in hospital nger than originally anticipated the family members begin to observe details of contemporary hospital life not noticed on occasional visits. What they perceive leads them to question those often heard, and taken-for-granted, values of efficiency, accountability and competitiveness. PMID- 11924624 TI - Drowning not waving: burnout and mental health nursing. AB - Mental health nursing as one of the human services professions has frequently, been associated with stress and burnout. The three dimensions of burnout; emotional exhaustion, depersonalisation and reduced personal accomplishment have major implications for the quality of the service that mental health nurses are capable of giving. Various studies have demonstrated mental health nurses suffer a high degree of burnout. Other studies have identified the stressors associated with mental health nursing that may be precursors to burnout. The Evaluation of the National Mental Health Strategy (1998) demonstrated that the Strategy was achieving many of its objectives. The objectives that have been achieved in service delivery have produced a work environment that has many of the identified stressors associated with burnout. Therefore, it is imperative that strategies are implemented that address the problem of stress and burnout in the current mental health system. PMID- 11924625 TI - The role of the psychiatric consultation-liaison nurse in the improved care of patients experiencing mental health problems receiving care within a general hospital environment. AB - The Psychiatric Consultation-Liaison Nurse (PCLN) has an imporrtant role in supporting nurses and other health care professionals in caring for patients experiencing mental health problems in a non-psychiatric environment. The limited available research findings suggest the PCLN role to be effective in terms of staff satisfaction, and some suggestion of cost savings is apparent. This paper refers to a survey of the attitudes of health professionals (n-113) too the services provided by the PCLN in a large metropolitan hospital in Melbournre, Australia. The results indicate a high level of satisfaction with the services of the PCLN. Enhancement of the participants' professional role, the professional role of nursing and improving health outcomes was amongst the perceived benefits of the PCLN role. PMID- 11924627 TI - A comparison of a traditional and an accelerated basic nursing education program. AB - In this study, the grade point average of graduates of a traditional three-year and an accelerated two-year pre-service nursing education program was compared, using comparison points of the divergence and the finish of the two programs. Results showed that while the accelerated program students had a significantly higher grade point average at the point of divergence, by the end of the two programs there was no significant difference. The accelerated program resulted in academically equivalent graduates in a shorter time but graduates paid a price in terms of stress and under-achievements. PMID- 11924626 TI - An Australian study on the sociocultural context of menopause: directions for contemporary nursing practice. AB - This research project by Berger (1997) investigates the physical, psychological, and sociocultural menopause experiences of a group of seventy women aged 45-70 years from Brisbane, Australia. Thus far the narrative provided for mid-life women shows a preoccupation with medical/biological perspectives that emphasize negative images of menopausal women as estrogen deficient and diseased. This new study is considered to be one of the few more recent attempts that investigates menopausal women within a sociocultural context and explores women's views about menopause as a normal, natural transition part of the ageing process. A qualitative approach (comprising focus groups and interviews) was aimed at women in mid-life who were willing to share their insights of this complex phase, to document individual variations and broad patterns. Findings provided enhanced understanding of many menopause aspects (such as appropriate treatment) that remain controversial. Content analysis uncovered three emerging themes: 1) contrary to popular beliefs menopause usually signified wellness; 2) apart from biological changes, the sociocultural context played a pivotal role in modifying menopause experiences; and 3) women did not feel adequately supported by health professionals. The findings signify that nurses are ideally placed to assist menopausal women in a variety of health care settings. PMID- 11924628 TI - Pharmacy compounding: customizing prescription drugs. PMID- 11924629 TI - Avoiding the hazards of medical gases. PMID- 11924630 TI - Trying to look SUNsational? Complexity persists in using sunscreens. PMID- 11924631 TI - Bone marrow transplants come of age. New hope for deadly diseases. PMID- 11924632 TI - Overcoming juvenile diabetes with a little planning and high-tech tools. PMID- 11924633 TI - Scheme to sell gas grill igniters for pain relief backfires. PMID- 11924634 TI - The pill at 40. PMID- 11924635 TI - First antibiotic in new class of drugs fights resistant infections. PMID- 11924636 TI - Light sabers protect sight of people with age-related eye disorder. PMID- 11924637 TI - Contaminated antiseptic products recalled. PMID- 11924638 TI - Medical device treats female sexual arousal disorder. PMID- 11924639 TI - High-tech hope for heartburn sufferers. PMID- 11924640 TI - Plans for more rigorous review of bioengineered foods. PMID- 11924641 TI - Sharks get cancer too. PMID- 11924642 TI - Studies show antioxidants no antidote for disease. PMID- 11924643 TI - Saline breast implants stay on market as experts warn about risks. PMID- 11924644 TI - Distribution of polyketide synthase genes in bacterial populations. PMID- 11924645 TI - A retrospective analysis of two independent prospective cartilage repair studies: autogenous perichondrial grafting versus subchondral drilling 10 years post surgery. AB - BACKGROUND: Experimental data indicate that perichondrial grafting to restore articular cartilage defects will result in repair with hyaline-like cartilage. In contrast. debridement and drilling results in repair with fibro-cartilage. In this retrospective study the long-term clinical results of both procedures were compared to evaluate the theoretical benefit of repair with hyaline-like tissue. METHODS: From two independent studies patients were selected with a cartilage defect in their knee. The selection was performed using strict inclusion criteria published elsewhere [Bouwmeester et al. Int. Orthop. 21 (1997) 313]. The patients were treated with either a perichondrium transplantation (PT group, n = 14) or with an 'open' debridement and drilling procedure (DD group, n = 11). The results of both procedures after 10-11 years were evaluated using the Hospital for Special Surgery Knee Score (HSSS), X-ray examination, clinical examination and visual analogue scales (VAS) for pain during walking and at rest. RESULTS: Both procedures resulted in a general improvement compared to the situation before the operation. After an average of 10 years in the PT group there were three failures, in the DD group none, success rates were 78% and 100%, respectively. When comparing the successful PT patients with the DD patients, there were no differences in HSSS and VAS data. Both groups showed an equal number of irregular operation surface sites on X-ray (PT 9/11 versus DD 8/10). CONCLUSIONS: This study shows that clinically at 10 years follow-up no difference was observed between debridement and drilling and perichondrium transplantation for treatment of an isolated cartilage defect. This raises questions about ongoing research to develop methods in order to improve the results of debridement and drilling as therapy for an isolated cartilage defect in a young patient (< or = 40 years). PMID- 11924646 TI - Biomechanical regulation of type I collagen gene expression in ACLs in organ culture. AB - In this study, an ex vivo organ culture system that allows the application of controlled loads to the anterior cruciate ligament (ACL) was designed and used to characterize the influence of a step input in mechanical load on gene expression. A procedure for isolating bone-ACL-bone (B-ACL-B) complexes from rat knees was developed. After harvest and 24 hour culture, B-ACL-B complexes exhibited percentages of viability similar to that in intact ACLs (approximately 90%). Application of a physiologically relevant load of 5 N (superimposed on a I N tare load) resulted in changes in levels of mRNA encoding type I collagen. While levels of type I collagen mRNA significantly increased 32+/-13% (mean +/- standard errors of the mean (SEM)) over controls within the first hour of loading, levels decreased significantly to 44+/-9% of control after 2 h. Displacements induced by the 5 N load were measured by video dimensional analysis. Calculated axial strains of 0.141+/-0.034 were achieved rapidly during the first hour and remained essentially unchanged thereafter. These results demonstrate the feasibility of maintaining ligaments in organ culture and illustrate the time course expression of type I collagen following the application of a mechanical load. PMID- 11924647 TI - Endogenous norepinephrine regulates blood flow to the intact rat tibia. AB - The goal of our study was to determine if endogenous norepinephrine (NE) has a role in the regulation of basal blood flow to intact bone. The experimental plan was to measure bone blood flow before and after pharmacological blockade of alpha adrenergic receptors. A significant increase in blood flow after receptor blockade would suggest that endogenous norepinephrine exerts a tonic constrictor effect on the vessels supplying blood to the bone. Mature, male rats were anesthetized with Inactin. Arterial blood pressure and left tibia blood flow (laser Doppler flowmetry) were measured. A cannula was inserted into the right iliac artery and advanced to the aortic bifurcation to deliver drugs into the left hindlimb circulation, including the left tibia vasculature. Bolus injection of norepinephrine caused a dose-dependent decrease in bone blood flow (30-40%). Blockade of alpha-adrenergic receptors with phentolamine or phenoxybenzamine attenuated by more than 50% the norepinephrine-induced decrease in bone blood flow. In separate rats that had not received exogenous norepinephrine, injection of phentolamine alone decreased bone vascular resistance by 34+/-3%. Similarly, phenoxybenzamine decreased resistance by 25+/-4%. These results are consistent with the conclusion that alpha-adrenergic receptors mediate a significant constriction of blood vessels which participate in the partial control of basal blood flow to the intact rat tibia. PMID- 11924648 TI - Kallikreins and kininogens in saliva and plasma of patients presenting with rheumatoid arthritis. AB - We evaluated the activities of salivary kallikrein and tissue/plasma kallikreins, and the plasma levels of high-molecular (HKg) and low-molecular (LKg) weight kininogens in patients with rheumatoid arthritis. The patients exhibited higher levels of the active salivary kallikrein compared to the controls. In contrast, the total salivary kallikrein activity of patients was not different from controls. In plasma from the patients, tissue kallikrein activity or plasma prekallikrein activity was not significantly different from the controls. Plasma HKg levels observed in patients were higher than in controls, whereas plasma LKg levels did not differ significantly from controls. Our results showed that most of the salivary kallikrein seen in patients is in its active form, suggesting the presence of systemic or local factors with the ability to activate salivary pro kallikrein. PMID- 11924649 TI - Anti-neutrophil cytoplasmic autoantibody (ANCA) profiles in propylthiouracil induced lupus-like manifestations in monozygotic triplets with hyperthyroidism. AB - We describe the ANCA profile of two monozygotic triplets (A and B) treated with propylthiouracil (PTU) for hyperthyroidism who developed LE-like manifestations. Triplet C also developed hyperthyroidism but was not treated with PTU and never experienced LE-like symptoms. Triplet A and B showed a marked rise in P-ANCA titer to 1:1280 after PTU was introduced whereas triplet C never had a titer higher than 1:80. Consecutive sera were investigated for ANCA to six different neutrophil granule proteins. Triplet A and B, but not C, both developed a strongly positive elastase-ANCA. Our results confirm the importance of a genetic factor influencing the susceptibility to drug-induced LE. PMID- 11924650 TI - Intervertebral discs which cause low back pain secrete high levels of proinflammatory mediators. AB - Herniated intervertebral disc tissue has been shown to produce a number of proinflammatory mediators and cytokines, but there have been no similar studies using discs from patients with discogenic low back pain. We have compared the levels of production of interleukin-6 (IL-6), interleukin-8 (IL-8) and prostaglandin E2 (PGE2) in disc tissue from patients undergoing discectomy for sciatica (63) with that from patients undergoing fusion for discogenic low back pain (20) using an enzyme-linked immunoabsorbent assay. There was a statistically significant difference between levels of production of IL-6 and IL-8 in the sciatica and low back pain groups (p < 0.006 and p < 0.003, respectively). The high levels of proinflammatory mediator found in disc tissue from patients undergoing fusion suggest that production of proinflammatory mediators within the nucleus pulposus may be a major factor in the genesis of a painful lumbar disc. PMID- 11924651 TI - Genetic susceptibility in Dupuytren's disease. TGF-beta1 polymorphisms and Dupuytren's disease. AB - Dupuytren's disease is a benign fibroproliferative disease of unknown aetiology. It is often familial and commonly affects Northern European Caucasian men, but genetic studies have yet to identify the relevant genes. Transforming growth factor beta one (TGF-beta1) is a multifunctional cytokine which plays a central role in wound healing and fibrosis. It stimulates the proliferation of fibroblasts and the deposition of extracellular matrix. Previous studies have implicated TGF-beta1 in Dupuytren's disease, suggesting that it may represent a candidate susceptibility gene for this condition. We have investigated the association of four common single nucleotide polymorphisms in TGF-beta1 with the risk of developing Dupuytren's disease. A polymerase chain reaction-restriction fragment length polymorphism method was used for genotyping TGF-beta1 polymorphisms. DNA samples from 135 patients with Dupuytren's disease and 200 control subjects were examined. There was no statistically significant difference in TGF-beta1 genotype or allele frequency distributions between the patients and controls for the codons 10, 25, -509 and -800 polymorphisms. Our observations suggest that common TGF-beta1 polymorphisms are not associated with a risk of developing Dupuytren's disease. These data should be interpreted with caution since the lack of association was shown in only one series of patients with only known, common polymorphisms of TGF-beta1. To our knowledge, this is the first report of a case-control association study in Dupuytren's disease using single nucleotide polymorphisms in TGF-beta1. PMID- 11924652 TI - Survival analysis of human meniscal transplantations. AB - We describe a prospective survival analysis of 63 consecutive meniscal allografts transplanted into 57 patients. The lateral meniscus was transplanted in 34, the medial meniscus in 17, and both menisci (combined) in the same knee in six. For survival analysis we used persistent pain or mechanical damage as clinical criteria of failure. A total of 13 allografts failed (5 lateral, 7 medial, 1 medial and lateral). A significant negative correlation (p = 0.003) was found between rupture of the anterior cruciate ligament (ACL) and successful meniscal transplantation. A significant difference (p = 0.004) in the clinical results was found between lateral and medial meniscal transplants. The cumulative survival rate of the lateral, medial and combined allografts in the same knee, based on the life-table method and the Kaplan-Meier calculation, was 76%, 50% and 67%, respectively. The survival of medial meniscal allografts may improve when reconstruction of the ACL is carried out at the same time as meniscal transplantation in an ACL-deficient knee. PMID- 11924653 TI - Anatomical reconstruction and Evans tenodesis of the lateral ligaments of the ankle. Clinical and radiological findings after follow-up for 15 to 30 years. AB - In this retrospective study, we assessed the outcome in 99 patients who underwent reconstruction of the lateral ligaments of the ankle for chronic anterolateral instability with a minimum follow-up of 15 years. Two techniques were compared: 54 patients had an anatomical reconstruction (AR group) and 45 had an Evans tenodesis (ET group). They were followed up for 19.9+/-3.6 years and 21.8+/-4.6 years, respectively. During follow-up, seven patients in the AR group and 17 in the ET group required a further operation (p = 0.004). At follow-up, significantly more patients (n = 15) in the ET group had limited dorsiflexion than in the AR group (n = 6, p = 0.007) and 18 in the ET group had a positive anterior drawer test compared with seven in the AR group (p = 0002). In the ET group 27 had tenderness on palpation of the ankle compared with 15 in the AR group (p = 0.001). Stress radiographs showed ligamentous laxity significantly more often in the ET group (n = 30) than in the AR group (n = 13, p < 0.001). The mean values for talar tilt and anterior talar translation were significantly higher in the ET group than in the AR group (p < 0.001, p = 0.007, respectively). There were degenerative changes on standard radiographs in 32 patients in the AR group and 35 in the ET group (p = 0.05). Four patients in the ET group had developed severe osteoarthritis compared with none in the AR group (p = 0.025). Assessment of functional stability revealed a mean Karlsson score of 83.7+/-10.4 points in the AR group and 67.0+/-15.8 points in the ET group (p < 0.001). According to the Good rating system, 43 patients in the AR group had good or excellent results compared with 15 in the ET group (p < 0.001). Compared with anatomical reconstruction, the Evans tenodesis does not prevent laxity in a large number of patients. Long-standing ligamentous laxity leads to degenerative change in the ankle, resulting in chronic pain, limited dorsiflexion and further operations. The functional result deteriorates more rapidly after the Evans tenodesis than after anatomical reconstruction. PMID- 11924654 TI - Quiz page. Urinary obstruction. PMID- 11924655 TI - Left ventricular assist device. PMID- 11924656 TI - Left ventricular assist device. PMID- 11924657 TI - Left ventricular assist device. PMID- 11924658 TI - Goal-directed therapy for severe sepsis. PMID- 11924659 TI - Goal-directed therapy for severe sepsis. PMID- 11924660 TI - An outbreak of primary pneumonic tularemia. PMID- 11924661 TI - Will parity in coverage result in better mental health care? PMID- 11924662 TI - Simple intervention to improve detection of child abuse in emergency departments. AB - PROBLEM: Child abuse is easily overlooked in a busy emergency department. DESIGN: Two stage audit of 1000 children before and after introduction of reminder flowchart. BACKGROUND AND SETTING: An emergency department in a suburban teaching hospital seeing about 4000 injured preschool children a year. KEY MEASURES FOR IMPROVEMENT: Number of records in which intentional injury was adequately documented and considered and the number of children referred for further assessment before and after introduction of reminder flowchart into emergency department notes. STRATEGIES FOR CHANGE: Nurses were asked to insert a reminder flowchart for assessing intentional injury into the notes of all children aged 0 5 years attending the department with any injury and to record the results of checking the child protection register. EFFECT OF CHANGE: Documentation of all eight indicators that intentional injury had been considered had increased in the second audit. Records of compatibility of history with injury and consistency of history increased from less than 2% to more than 70% (P<0.0001). More children were referred for further assessment in the second audit than the first, although the difference was not significant (6 (0.6%) v 14 (1.4%), P=0.072). The general level of awareness and vigilance increased in the second audit, even for children whose records did not contain the flowchart. LESSONS LEARNT: Inclusion of a simple reminder flowchart in the notes of injured preschool children attending the emergency department increases awareness, consideration, and documentation of intentional injury. Rates of referral for further assessment also increase. PMID- 11924663 TI - Evidence base of clinical diagnosis: Rational, cost effective use of investigations in clinical practice. PMID- 11924664 TI - Bullying in medicine. Both sides need help when bullying happens. PMID- 11924665 TI - Bullying in medicine. We must all learn from our unacceptable behaviour. PMID- 11924667 TI - Redefining leadership. Successful leadership requires range of styles. PMID- 11924668 TI - Redefining leadership. Hospital service needs doctors who are better educated and trained. PMID- 11924669 TI - Violence in the workplace. Mentally ill patients are responsible for their actions. PMID- 11924670 TI - Violence in the workplace. Zero tolerance is not helpful in mental health. PMID- 11924671 TI - Statins as the new aspirin. National Institute for Clinical Excellence should assess statins. PMID- 11924672 TI - Full time forensic pathology service. Role of clinical necropsy in medical practice needs to be strengthened. PMID- 11924673 TI - Why conservationists should heed Pokemon. PMID- 11924674 TI - Assessing the megatons to megawatts program. PMID- 11924675 TI - Assessing the megatons to megawatts program. PMID- 11924676 TI - Aristotle's view of a creator. PMID- 11924677 TI - Changes in organ perfusion and weight ratios in post-simulated microgravity recovery. AB - Head-down tilt models have been used as ground-based simulations of microgravity. Our previous animal research has demonstrated that there are significant changes in fluid distribution within 2 h after placement in a 45 degrees head-down tilt (45HDT) position and these changes in fluid distribution were still present after 14 days of 45HDT. Consequently, we investigated changes in fluid distribution during recovery from 16 days of 45HDT. Changes in radioactive tracer distribution and organ/body weight ratio were examined in rats randomly assigned to a 45HDT or prone control group. The 45HDT rats were suspended for 16 days and then allowed to recover at the prone position 0, 77, 101, or 125 h post-suspension. Animals were injected with technetium-labeled diethylenetriamine pentaacetate (99mTcDTPA, MW=492 amu, physical half-life of 6.02 h) and then killed 30 min post-injection. Lungs, heart, liver, spleen, kidneys, and brain were harvested, weighed, and measured for radioactive counts. Statistical analyses included two-way analysis of variance (ANOVA) that compared 45HDT versus controls at the four experimental time points. The organ weight divided by the body weight ratio for the brain, heart, kidneys and liver in the 45HDT rats was significantly different than the control rats, regardless of time (treatment). There was no difference between the different time points (time). The average 99mTcDTPA count divided by the organ weight ratio values for the heart, liver, and spleen were significantly higher in the 45HDT group than the control group. The average counts for the heart and spleen were significantly higher at 77, 101, and 125 h than at time zero. We conclude that the major organs have different recovery patterns after 45HDT for 16 days in the rat. PMID- 11924678 TI - Changes in mood status and neurotic levels during a 20-day bed rest. AB - This study evaluated changes of mood status and depressive and neurotic levels in nine young male subjects during a 20-day 6 degrees head-down tilting bed rest and examined whether exercise training modified these changes. Participants were asked to complete psychometrical inventories on before, during, and after the bed rest experiment. Depressive and neurotic levels were enhanced during bed rest period according to the Japanese version of Zung's Self-rating Depression Scale and the Japanese version of the General Health Questionnaire. Mood state "vigor" was impaired and "confusion" was increased during bed rest and recumbent control periods compared to pre-bed rest and ambulatory control periods according to the Japanese version of Profiles of Mood State, whereas the mood "tension-anxiety", "depression-dejection", "anger-hostility" and "fatigue" were relatively stable during experiment. Isometric exercise training did not modify these results. Microgravity, along with confinement to bed and isolation from familiar environments, induced impairment of mental status. PMID- 11924679 TI - Repair of the achilles tendon rupture: a functional comparison of three surgical techniques. AB - A retrospective comparison of three surgical procedures for the acute treatment of closed ruptures of the Achilles tendon is presented: 1) open repair, 2) a percutaneous technique, and 3) a combined mini-open and percutaneous technique. The authors compared the results of 52 tendon ruptures, including 15 open repairs, 15 percutaneous repairs, and 22 combined repairs, with minimal follow-up of 12 months. The authors evaluated the parameters of strength, performance, use of shoes, time of returning to work, range of ankle motion, calf circumference, pain, ability to perform 20 toe-raises on each side, and MRI findings. In comparison to the uninjured leg, the strength of the involved extremity was 74% in the patients with an open repair, 88% in patients with a percutaneous repair, and 92% in the combined mini-open and percutaneous procedure. The combined mini open and percutaneous repair gave significantly better results than the other surgical procedures. On MRI, the area of the posterior calf of the injured leg, in comparison to the contralateral uninjured leg was 82% in open repair, 81% in the percutaneous procedure, and 91% in combined mini-open and percutaneous procedure. These differences were found to be statistically significant (p < .01). On the basis of their results, the authors prefer to perform the combined percutaneous and mini-open repair of Kakiuchi for the repair of acute Achilles tendon ruptures. PMID- 11924680 TI - A comparison of soft-tissue anchors in tendo achilles reattachment. AB - This prospective study evaluated four soft-tissue fixation modalities, used in seven different combinations, to reattach the tendo Achilles in 34 cadaveric specimens. Ultimate loads, elastic moduli, and modes of failure were evaluated by loading the specimen in a cantilevered fashion on an Instron. Mann-Whitney U tests were performed to compare the failure load data for statistical significance. Although the use of two Mitek SuperAnchors showed better load resistance than one anchor (p < .01), there was no significant improvement between using two or three anchors (one anchor 116 +/- 24 N, two anchors 234 +/- 21 N, three anchors 277 +/- 80 N). Two Bionx Bankart Tacks demonstrated no significant difference over using a single tack (one tack 178 +/- 57 N, two tacks 214 +/- 86 N). No statistical difference was observed between the screw and washer systems (screw with polyacetal resin washer 307 +/- 80 N, screw with metal washer 290 +/- 81 N). Both screw and washer systems did show greater stability when compared with a single Mitek SuperAnchor (p < .01) or a single Bionx Bankart Tack (p < .05). Similar analyses using the Mann-Whitney U tests were performed on the elastic modulus data. Analysis of the displacement data among all groups showed no statistical difference. Observations of the mode of failure exhibited 86% of Mitek SuperAnchor failed secondary to suture, and 70% of the Bionx Bankart Tack and 90% of the screw and washer systems failed because of the tendon shearing around the fixation. The comparisons of cost-effectiveness among the fixations showed the Synthes screw and polyacetal resin spiked washer to have the lowest cost to load ratio ($0.15/N). PMID- 11924681 TI - First metatarsophalangeal joint arthrodesis: quantitative mechanical testing of six-hole dorsal plate versus crossed screw fixation in cadaveric specimens. AB - Quantitative strength analysis of first metatarsophalangeal joint arthrodesis was performed using two fixation techniques: a small 6-hole plate with an interfragmentary screw or two crossed lag screws. Twelve matched-pair fresh frozen cadaveric specimens (24 trials) were used for direct comparison of each of the two fixation techniques. All joint surfaces were prepared with power conical reamers utilizing a standard technique. The fixation construct was stressed to failure on each specimen using a computer-integrated materials tester. Fixation stiffness defined as force (load) over displacement and point of ultimate failure was evaluated. The six-hole plate and interfragmentary screw fixation method was a statistically stiffer form of fixation (p > .01) and displayed a greater point of ultimate failure (p > .002) under the laboratory conditions. PMID- 11924682 TI - Predislocation syndrome: a review and retrospective analysis of eight patients. AB - Predislocation syndrome is defined as an either an acute, subacute, or chronic inflammatory process involving the lesser metatarsophalangealjoints. If left untreated, this process can lead to plantarplate and capsular attenuation and metatarsophalangeal joint instability. A retrospective analysis were performed on eight patients who were treated with a flexor tendon transfer for this condition. A subjective analysis and chart review were performed to determine the final outcome of these patients. All patients had a chief complaint of a painful second metatarsophalangeal joint with seven of eight patients having an associated hallux valgus deformity. Excellent results were achieved in six patients. Residual stiffness was the primary complaint of two patients following surgery, suggesting that metatarsophalangeal joint stiffness may be a potential problem after flexor digitorum longus tendon transfers. However, we concluded that the flexor digitorum longus tendon transfer remains an excellent procedure for second toe instability and late-stage predislocation syndrome. A literature review describing the epidemiology, symptom complex, physical findings, radiographic signs and therapies used to manage predislocation syndrome were also discussed. PMID- 11924683 TI - Verrucous carcinoma of the foot: a review and report of two cases. AB - Verrucous carcinoma is an uncommon form of squamous cell carcinoma. Clinical and histological features of this condition are reviewed. Excision is the treatment of choice due to local aggressiveness and infrequent metastasis. Two cases involving the foot are presented as well as a survey of previously documented cases of this condition. PMID- 11924684 TI - Complex regional pain syndrome: a report of two cases recalcitrant to usual treatment protocols. AB - In this report the authors present a review of complex regional pain syndrome and two case reports of complex regional pain syndrome that were recalcitrant to the usual treatments. The first case presented is a middle-aged female who developed signs and symptoms of post-traumatic complex regional pain syndrome. The second case is a woman with a pre-existing history of complex regional pain syndrome whose condition worsened after surgery despite appropriate perioperative precautions. These cases are unique because in both cases an early diagnosis of complex regional pain syndrome was established, yet they were both resistant to the usual treatment protocols. PMID- 11924686 TI - Functional instability of the ankle and the role of neuromuscular control: a comprehensive review. AB - A review of current knowledge of the clinical syndrome of functional ankle instability is presented. Recent evidence has demonstrated that the majority of patients with functional instability of the ankle do not have mechanical hypermobility of the ankle joint. Functional instability of the ankle results from a loss of neuromuscular control. Components of neuromuscular control include proprioception, muscle strength, muscle reaction time, and postural control. Proprioceptive deficits lead to a delay in peroneal reaction time, which appears to be a peripheral reflex. Proprioception and eversion muscle strength improve with the use of passive supportive devices. Balance and postural control of the ankle appear to be diminished after a lateral ankle sprain and can be restored through training that is mediated through central nervous mechanisms. Methods of detecting deficits in neuromuscular control are presented along with rehabilitation techniques to treat functional instability of the ankle. PMID- 11924685 TI - Neurilemoma of the medial plantar nerve: a case report. AB - The incidence of neurilemoma in the foot is uncommon. Neurilemoma of the medial plantar nerve distal to the tarsal tunnel, with symptoms isolated to the foot, is rare. A case presentation of neurilemoma arising from the distal portion of the medial plantar nerve that was identified and excised is presented with a 19-month follow-up. PMID- 11924687 TI - Fibular groove deepening for recurrent peroneal subluxation. PMID- 11924688 TI - The single K-wire fixation of the Mitchell bunionectomy. PMID- 11924689 TI - Increasing upper level slots. PMID- 11924690 TI - Suicide prevention: broadening the field toward a public health approach. PMID- 11924691 TI - Factors associated with the medical severity of suicide attempts in youths and young adults. AB - This study examined factors associated with the medical severity of suicide attempts focusing on demographic characteristics, mental health characteristics, and the circumstances of the suicide attempt. Analyses were based on 153 nearly lethal suicide attempters and 47 less lethal suicide attempters aged 13-34 years who presented to emergency departments in Houston, Texas. The results show that young age was significantly associated with a nearly lethal suicide attempt. Prior suicide attempts, hopelessness, depression, and help-seeking (ever) were significantly and negatively associated with a nearly lethal suicide attempt. None of the suicide attempt factors occurring prior to the attempt were associated with a nearly lethal suicide attempt. PMID- 11924692 TI - Interviewing suicide "decedents": a fourth strategy for risk factor assessment. PMID- 11924693 TI - Alcohol consumption and nearly lethal suicide attempts. AB - We conducted a case-control study of the association between nearly lethal suicide attempts and facets of alcohol consumption; namely, drinking frequency, drinking quantity, binge drinking, alcoholism, drinking within 3 hours of suicide attempt, and age began drinking. Subjects were 13-34 years of age. In bivariable analyses, all measures were associated with nearly lethal suicide attempts. Odds ratios ranged from 2.4 for alcoholism to 7.0 for drinking within 3 hours of attempt. All exposure variables except age began drinking exhibited a J-shaped relationship between alcohol exposure and nearly lethal suicide attempt. After controlling for potential confounders and other measures of alcohol exposure, drinking within 3 hours of attempt remained most strongly (odds ratios > 6) associated. Alcoholism remained significantly associated in most models, but at lower strength. PMID- 11924694 TI - The influence of geographic mobility on nearly lethal suicide attempts. AB - Teenagers and young adults are very mobile and mobility has been identified as a potential risk factor for suicidal behavior. We conducted a population-based, case-control study of nearly lethal suicide attempts with 153 cases and 513 controls. Study participants were asked about changing residence over the past 12 months. Results indicate that moving in the past 12 months is positively associated with a nearly lethal suicide attempt (adjusted odds ratio of 2.1, with 95% confidence interval of 1.4-3.3), as are specific characteristics of the move (e.g., frequency, recency, distance, and difficulty staying in touch). These findings confirm and extend prior ecologic research by demonstrating a relationship, at the individual level, between the geographic mobility of adolescents and young adults and nearly lethal suicide attempts. PMID- 11924695 TI - Characteristics of impulsive suicide attempts and attempters. AB - Suicide attempts often are impulsive, yet little is known about the characteristics of impulsive suicide. We examined impulsive suicide attempts within a population-based, case-control study of nearly lethal suicide attempts among people 13-34 years of age. Attempts were considered impulsive if the respondent reported spending less than 5 minutes between the decision to attempt suicide and the actual attempt. Among the 153 case-subjects, 24% attempted impulsively. Impulsive attempts were more likely among those who had been in a physical fight and less likely among those who were depressed. Relative to control subjects, male sex, fighting, and hopelessness distinguished impulsive cases but depression did not. Our findings suggest that inadequate control of aggressive impulses might be a greater indicator of risk for impulsive suicide attempts than depression. PMID- 11924696 TI - Medical conditions and nearly lethal suicide attempts. AB - Physical illness has been studied as a risk factor for suicidal behavior, but little is known about this relationship among younger persons. We conducted a population-based, case-control study in Houston, Texas, from November 1992 through September 1995. The final sample consisted of 153 case- and 513 control subjects aged 13 to 34 years. Case patients were identified at hospital emergency departments and met criteria for a nearly lethal suicide attempt. Control subjects were recruited via a random-digit-dial telephone survey. Case patients were more likely than controls to report having any serious medical conditions (crude OR = 3.23; 95% CI = 2.12-4.91). After controlling for age, race/ethnicity, alcoholism, depression, and hopelessness, the adjusted odds ratio for men was 4.76 (95% CI = 1.87-12.17), whereas the adjusted odds ratio for women was 1.60 (95% CI-0.62-4.17), suggesting that young men with medical conditions are at increased risk for nearly lethal suicide attempts. Increased efforts to identify and appropriately refer these patients are needed. PMID- 11924697 TI - Help-seeking behavior prior to nearly lethal suicide attempts. AB - The association between help-seeking and nearly lethal suicide attempts was evaluated using data from a population-based, case-control study of 153 13- to 34 year-old suicide attempt case-patients treated at emergency departments in Houston, Texas, and a random sample of 513 control-subjects. Measures of help seeking included whether the participant sought help for health/emotional problems in the past month, type of consultant contacted, and whether suicide was discussed during the interaction. Overall, friends/family were consulted most frequently (48%). After controlling for potential confounders, case-patients were less likely than control-subjects to seek help from any consultant (OR = 0.5, 95% CI = 0.3-0.8) or a professional (e.g., physician, counselor) consultant (OR = 0.5, 95%CI = 0.29-0.8). Among those who sought help, case-patients were more likely than to discuss suicide (OR = 2.6, 95% CI = 1.2-5.4), particularly with professionals (OR = 11.8, 95% CI = 3.2-43.2). Our findings suggest that efforts to better understand the role of help-seeking in suicide prevention, including help sought from family and friends, deserves further attention. PMID- 11924698 TI - An unmatched case-control study of nearly lethal suicide attempts in Houston, Texas: research methods and measurements. AB - This article details the research methods and measurements used in conducting a population-based, case-control study of nearly lethal suicide attempts among persons aged 13-34 years, residing in Houston, Texas. From November 1992 to July 1995, we interviewed 153 case subjects presenting at one of three participating hospital emergency departments and used random digit dialing to identify 513 control subjects residing in the same catchment area in which cases were enlisted. Unlike most research in this area, this study was designed to extend our understanding of suicidal behavior and prevention activities beyond identification and treatment of depression and other mental illnesses. We discuss the overall strengths and weaknesses of our study design and conclude that this methodology is well suited for studying rare outcomes such as nearly lethal suicide. PMID- 11924699 TI - Studying survivors of nearly lethal suicide attempts: an important strategy in suicide research. AB - The series of articles in this special issue of SLTB from the Centers for Disease Control and Prevention (CDC) study of nearly lethal suicide attempts represents an important contribution to research on suicidal behavior. The investigative approach used is attractive, yet also challenging in terms of research methodology. In this commentary I take the opportunity to highlight certain aspects of this line of research as well as to comment on the specific findings of the CDC study and their relationship to existing knowledge. I also discuss the future potential for this research approach and further questions that might be addressed by it. PMID- 11924700 TI - Determination of firing distance using the rhodizonate staining technique. AB - The histological staining technique using rhodizonate is also effective for the determination of the firing distance by examining the distribution and intensity of the staining reaction. The differentiation between absolute close-range shots and long-range shots is generally possible without any doubt. The method is not recommended for routine examinations but it is very useful for cases lacking the possibility to investigate smoke and powder deposits in a criminalistic manner, i.e. surgical skin biopsies of hospitalised victims and skin highly altered by the effects of fire, water or by post-mortem decomposition. PMID- 11924701 TI - Preliminary approach to elucidate the role of pigment as a binding site for drugs and chemicals in anagen hairs: pigments as carriers for 3H-haloperidol in HaCaT/Sk-Mel-1 co-cultures. AB - In view of the melanin-binding characteristics of haloperidol and its differential uptake by pigment- and non-pigment-producing cells, a co-culture of HaCaT with Sk-Mel-1 cell lines was performed to investigate whether melanosomes act as carriers for drug molecules associated with the pigments. Initially, HaCaT and Sk-Mel-1 cells were separately cultivated in the presence of 3H-haloperidol (400 pmol/ml medium ) for 28 days followed by subsequent co-cultivation in the absence of 3H-haloperidol for 5 days. The transfer of pigments into the keratinocytes during co-culture was confirmed by transmission electron microscopy. After the co-culture experiments a striking increase (> or = 50%) of 3H-haloperidol was observed in the pigmented HaCaT cells compared to the unpigmented keratinocytes. The present study proved the role of pigments as carriers for melanin-associated drug molecules. The results supported the hypothesis that hair pigment might be a factor affecting the outcome of hair assays for particular categories of commonly used licit and illicit substances. The chosen cell lines and the developed co-culture system may represent suitable in vitro models to study differential drug uptake into cell populations present in the skin or in the growing hair follicle as well as to elucidate drug uptake due to melanocyte-keratinocyte interactions. PMID- 11924702 TI - Detection of cardiomyocyte apoptosis in forensic autopsy cases. AB - The purpose of the present study was to determine reliable parameters for the detection of apoptotic cells for use as a diagnostic marker during the early stage of acute myocardial infarction (AMI) in forensic autopsy cases. Myocardial tissues taken from forensic autopsy cases were examined by immunohistochemical and molecular-biological methods using the terminal deoxynucleotidyl transferase mediated dUTP biotin nick end-labelling (TUNEL) and the DNA laddering methods. In cases of AMI with a time period between 2 h from onset to death and 20 h post mortem time, the nuclei of cardiomyocytes were stained positive with the TUNEL method and DNA fragmentation of myocardial cells was detected by agarose gel electrophoresis. Similar findings were obtained in cases of carbon monoxide (CO) intoxication. However, no apoptotic cells were found in other cases such as methamphetamine (MAP) intoxication, tetrodotoxin intoxication, alcohol intoxication, asphyxia, head injury, heart injury or myocarditis. These findings suggested that it would be possible to apply TUNEL-positive cells as a diagnostic marker during the early stages of AMI. PMID- 11924703 TI - TUNEL: a useful screening method in sudden cardiac death. AB - The primary objective of this study was to investigate if detection of apoptosis in the heart can be used to diagnose early myocardial ischaemia. The material consisted of myocardial tissue from autopsy cases: 10 cases with occlusive, thrombotic coronary artery disease and acute myocardial infarction, 10 cases of sudden cardiac death without coronary artery disease (CAD) and 8 controls without cardiovascular disease and with known causes of death. Necrotic changes in the myocardium were detected with hematoxylin-erythrosin-saffron, Mallory's PTAH stain and with antibodies against complement 9. Apoptotic nuclei were visualised with two different kits using the terminal deoxynucleotidyl transferase-mediated desoxyuridinetriphosphate nick end-labeling (TUNEL) method on histological sections. In the patients with CAD, early myocardial infarction was found in one defined area of the ventricular wall; apoptotic myocyte nuclei were observed not in the necrotic lesions, but evenly spread usually without a gradient, all over the myocardium with a mean number per high power field of 29% (range 3-56%) of the total number of myocyte nuclei. In the sudden cardiac deaths without CAD, necrosis was scarce and distributed both focally and irregularly in both the left and right ventricular walls. With few exceptions, the percentage of apoptotic myocyte nuclei exceeded 20% in all sections (mean 24%, range 0-68%). No difference was seen between patients with CAD and those without CAD (p > 0.05). With the TUNEL method, positively stained nuclei were seen very early and extensively all over the myocardium. It is not certain that they represent true apoptosis induced by ischemia, but TUNEL appears to be a useful screening method in cases where sudden cardiac death is suspected. PMID- 11924704 TI - Dioxin in infants--an environmental hazard? AB - The concentrations of the most common dioxin and dibenzofuran congeners were measured in different tissues (e.g. liver, kidneys, subcutaneous fatty tissue and spleen) from 27 infants who died suddenly and unexpectedly. The cases could be subdivided into 2 groups consisting of 15 infants who died in 1991/1992 and in 12 infants who died in 1996/1997. The autopsies were carried out using a standardised protocol and additionally the parents were asked to supply details of the nutritional conditions. The age of the mother and the birth order of the infants were also recorded. From the results obtained by correlating these parameters with the dioxin concentrations three main factors could be established: 1) there was a significant decrease in the total dioxin concentration in infant tissues from 1991/1992 to 1996/1997 indicating a decrease in the environmental dioxin levels due to a decrease in dioxin emission, 2) the birth order was inversely and the duration of breast feeding directly proportional to the dioxin concentrations thus showing that the mothers can decontaminate themselves by breast feeding and 3) an accumulation of specific dioxin congeners was observed in the liver tissue but the pathophysiological significance of these observations is not yet fully understood. Because of the well-known beneficial effects of breast feeding and considering the results of the present study, this type of infant nutrition can be recommended without any restrictions. PMID- 11924705 TI - Immunohistochemical detection of an ABO-incompatible blood transfusion in formalin-fixed tissue. AB - A fatal incident of an ABO incompatible erythrocyte transfusion in a 75-year-old male patient who suffered from dilated cardiomyopathy with cardiac failure is reported. Blood group A red cells were transfused to the unintended recipient who had blood group O. The patient died 45 min after the incompatible erythrocyte transfusion. The way the incident happened remained unclear and the immunohistochemical detection of ABO incompatible erythrocytes in formaldehyde fixed paraffin-embedded kidney, lung, liver and spleen tissue provided the only material evidence of the transfusion error. PMID- 11924706 TI - A death resulting from inadvertent intravenous infusion of enteral feed. AB - A female patient suffering from the after-effects of an intracerebral hemorrhage, inadvertently received approximately 50 ml of enteral feed containing high molecular weight dextrin intravenously and died 6 h later despite intensive emergency resuscitation attempts. The total quantity of enteral feed received was calculated from the amounts of dextrin measured in the blood. This is the first report describing how the total quantity of enteral feed administered intravenously was determined using biochemical analysis. PMID- 11924707 TI - Paternity testing after pregnancy termination using laser microdissection of chorionic villi. AB - We report an unusual case of paternity testing from residues of chorionic villi 5 weeks after pregnancy termination. The autopsy of a 32-year-old female homicide victim revealed the presence of intact chorionic villi at the former placenta implantation site. Fetal cells were selectively isolated by laser-induced microdissection of the remaining villi to avoid contamination with maternal DNA. Simultaneous amplification of 12 STR loci in 2 PCR reactions resulted in a combined probability of paternity of 99.94%. This case demonstrates that laser assisted microdissection and multiplex STR typing provide tools for paternity testing performed on endometrial mucosa long after the product of conception was removed by therapeutic abortion. PMID- 11924708 TI - Sudden death due to rupture of the arteria pancreatica magna: a complication of an immature pseudocyst in chronic pancreatitis. AB - Massive haemorrhage due to rupture of single pancreatic or peripancreatic vessels is a very rare but potentially lethal complication of acute and chronic pancreatitis. The splenic, gastroduodenal, and pancreatoduodenal arteries are the more commonly involved vessels, and rupture occurs mostly as a complication of large mature pseudocysts. We report a sudden death due to massive bleeding caused by rupture of the great pancreatic artery (arteria pancreatica magna), a complication of a small immature pseudocyst, in a 49-year-old male alcoholic with inactive chronic pancreatitis. PMID- 11924709 TI - Confusing exit gunshot wound--"two for the price of one". AB - During the post-mortem examination of a 36-year-old shooting victim, two entrance gunshot wounds in the back and only one corresponding exit wound in the chest were detected. The likelihood of a retained fragmented or whole bullet, or an embolic phenomenon was eliminated by full body x-ray examination. The only remaining plausible explanation was that the two bullet tracts had converged within the thorax, eventually exiting through a common exit wound. Dissection of the projectiles' path confirmed this unusual phenomenon. Autopsy techniques for gunshot wound cases are discussed. PMID- 11924710 TI - Capillary zone electrophoresis and artificial neural networks for estimation of the post-mortem interval (PMI) using electrolytes measurements in human vitreous humour. AB - Determination of electrolyte concentrations (mainly potassium) in vitreous humour has long been considered an important tool in human death investigations for the estimation of the post-mortem interval (PMI). On the basis of its well known potential in ion analysis, capillary zone electrophoresis (CZE) has recently been applied to achieve a rapid and simultaneous determination of inorganic ions in this extracellular fluid. In the present work, artificial neural networks (ANN) were applied for modelling of the relationship of multicomponent CZE analysis of K+, NH4+, Na+, and Ba2+ ions in vitreous humour with PMI. In a study based on 61 cases with different causes of death and a known PMI ranging from 3 to 144 h, the use of ANNs considering all inorganic ion data from the human vitreous humour, achieved a substantial improvement of post-mortem interval prediction. Good linear correlation was observed (r2 = 0.98) and in comparison to the traditional linear least squares (LLS) method applied only to K+ levels in the vitreous humour, the prediction of PMI with ANN was improved by a factor of 5 from approximately +/- 15 h to less than 3 h. PMID- 11924711 TI - Maternal death in pregnancy from HELLP syndrome. A report of three medico-legal autopsy cases with special reference to distinctive histopathological alterations. AB - Maternal death from HELLP syndrome, a complication of (pre-) eclampsia during pregnancy or postpartum, is rarely encountered in forensic pathology. We report three cases of HELLP syndrome with fatal outcome putting the main focus on the histopathological features of the disease. We found an almost identical histopathological pattern in the liver (periportal coagulation necrosis, hepatic haemorrhages sharply demarcated by an extended fibrin network from the surrounding unaffected liver parenchyma, focal leukostasis in liver sinusoids and swelling of Kupffer's cells, absence of inflammatory cellular infiltrates in liver plates, lack of fatty transformation of hepatocytes) and kidneys (bloodless glomeruli with swollen and vacuolated intracapillary cells, cigar-shaped capillary loops, enlarged glomerular tufts with herniation of capillary loops into the proximal convoluted tubules, swelling of mesangial cells) in all three cases. The histopathological alterations in the liver and kidneys can be considered characteristic for the disease and their presence may enable the forensic pathologist to establish the definite post-mortem diagnosis of HELLP syndrome in questionable cases. PMID- 11924712 TI - Determination of chronic abuse of the anaesthetic agents midazolam and propofol as demonstrated by hair analysis. AB - A 44-year-old female nurse in a department of anesthesiology was found dead at home. An empty bottle of Hypnovel (midazolam 5 mg/5 ml) and a hypodermic syringe were found near the corpse. The nurse was a known abuser of anaesthetic agents for many years. A complete screening for general unknown substances by FPIA, GC/MS, head space GC/MS and HPLC/DAD revealed the simultaneous presence of midazolam, propofol and ethanol in femoral blood. Segmental analysis of a 6-cm long hair strand revealed the presence of midazolam and propofol in each 2-cm long segment. Repetitive consumption of the two anaesthetic agents during the last 6 months before the death was therefore demonstrated. These compounds were also detected in pubic and axillary hairs. Self-administration of midazolam and propofol without respiratory assistance and medical control certainly contributed to the death. PMID- 11924713 TI - Preliminary approach to elucidate the role of pigment as a binding site for drugs and chemicals in anagen hair: differential uptake of 3H-haloperidol by pigment producing compared to non-pigment-producing cell lines. AB - A striking difference was observed for cellular-bound drug in HaCaT and Sk-Mel-1 cells for a fixed drug exposure time of 72 h and varying 3H-haloperidol concentrations in the culture media. Drug uptake was dependent on drug concentration and linearly correlated for both the non-pigment- and the pigment producing cells which however was different in magnitude. In an additional investigation the time course of drug uptake during 3H-haloperidol exposure (400 pmol/ml; 28 days) revealed increasing drug concentrations in the Sk-Mel-1 population, whereas drug concentrations in the keratinocytes reached a plateau within a short time period. In contrast to the HaCaT cells no tendency to saturation was observed for the pigment-producing cell line. At the end of the experiments 3H-haloperidol concentrations in Sk-Mel-1 were found to be approximately tenfold higher than in HaCaT. PMID- 11924714 TI - Lack of association between Y chromosome Alu insertion polymorphism and hypertension. AB - There is an inherited paternal predisposition to hypertension. Y chromosome alphoid satellite variation was recently reported to be linked to diastolic blood pressure. To determine whether there is also a Y chromosome marker linked to hypertension, we investigated the prevalence of the Y chromosome Alu insertion polymorphism (YAP) at DYS287 and its association with hypertension in the Aomori population in the northern area of Honshu Island, Japan. YAP was present in 98 of 285 male residents and absent in the rest. The YAP prevalence in the present study would appear to suggest that the present study population represents the general male population in central Japan. Within the study population, there were 110 hypertensive subjects and 104 normotensive subjects. YAP frequency in the hypertensive subjects was not different from that in the normotensive subjects. These results suggest that the YAP is not likely to be a genetic-susceptibility factor for hypertension in the Aomori population. PMID- 11924715 TI - Novel mechanisms of the antiproliferative effects of amlodipine in vascular smooth muscle cells from spontaneously hypertensive rats. AB - The calcium channel blocker amlodipine continues to be of interest due to its potential proven ability to hinder the progression of atherosclerosis and reduce the number of clinical ischemic events. Vascular smooth muscle cells (VSMC) from spontaneously hypertensive rats (SHR) are useful in the study of atherosclerosis because they show exaggerated growth with production of angiotensin II (Ang II) by conversion to the synthetic phenotype. To clarify mechanisms of the antiproliferative effects of amlodipine, we evaluated effects of the expression of growth factors, the changes in phenotype, and the proliferation of VSMC from SHR. Amlodipine significantly inhibited basal DNA synthesis and proliferation of VSMC from SHR. Amlodipine also inhibited expression of platelet-derived growth factor (PDGF) A-chain, transforming growth factor beta1 (TGF-beta1) and basic fibroblast growth factor (bFGF) mRNAs in VSMC from SHR. Decreases in levels of PDGF A-chain and bFGF mRNAs in VSMC from SHR were greater with amlodipine than with nifedipine. Amlodipine significantly inhibited expression of the synthetic phenotype markers osteopontin and matrix Gla mRNAs, indicating that it inhibited the exaggerated growth of VSMC from SHR and suppressed the change from the contractile phenotype to the synthetic phenotype. Thus, amlodipine may be a beneficial therapeutic agent for patients with hypertensive vascular diseases. PMID- 11924716 TI - Discriminating factors for recurrent hypertension in patients with primary aldosteronism after adrenalectomy. AB - Patients with primary aldosteronism show relatively high rates of hypertension after adrenalectomy, but the risk factors for postoperative hypertension remain unclear. Forty-six patients with primary aldosteronism (PA) who had undergone adrenalectomy between 1976 and 1998 were enrolled in this study. Follow-up information including blood pressure (BP) and cardiovascular complications was collected by means of correspondence or telephone contact. At discharge BP was normalized in 34 patients (72%); hypertension persisted in the remaining 12 patients, but BP control was significantly improved. The patients who remained hypertensive at discharge had longer durations of hypertension than did those with normalized BP. After an average follow-up period of 12.2 years, 16 of 34 BP normalized patients (47%) had recurrent hypertension. Age at adrenalectomy, preoperative serum creatinine level and systolic blood pressure at discharge were significantly higher in patients with recurrent hypertension than in those without it. A multivariate logistic regression analysis revealed that only the level of serum creatinine was independently associated with the incidence of recurrent hypertension. Patients with serum creatinine of 0.9 mg/dl or greater had significantly higher rates of recurrent hypertension than those with lower values of serum creatinine. Cardiovascular complications occurred in 5 patients prior to the surgery and in 2 patients during the follow-up period. Although the severity of renal involvement is subclinical, renal damage may play an important role in the development of hypertension during a long period after adrenalectomy in patients with PA. PMID- 11924717 TI - Continuous blockade of L-type Ca2+ channels suppresses activation of calcineurin and development of cardiac hypertrophy in spontaneously hypertensive rats. AB - We examined whether Ca2+ channel blockers inhibit the activation of the Ca2+ dependent phosphatase calcineurin and the development of cardiac hypertrophy in spontaneously hypertensive rats (SHR). We randomly divided 12-week-old SHR into three groups, one each receiving vehicle, bolus injection or continuous infusion of nifedipine (10 mg/kg/day) from 12 to 24 weeks of age. Systolic blood pressure (BP) and heart rate were measured every week after the treatment using the tail cuff plethysmography method. After 4, 8 and 12 weeks of treatment, 6 rats of each group were subjected to examinations that included an assay for calcineurin activity in the heart, magnetic resonance imaging (MRI), histology and Northern blot analysis. Continuous infusion of nifedipine consistently reduced BP, whereas bolus injection resulted in a fluctuation of BP. Continuous infusion of nifedipine not only reduced left ventricular mass but also decreased the transverse diameter of cardiomyocytes, interstitial fibrosis and the expression of the atrial natriuretic peptide and brain natriuretic peptide genes in the heart, while bolus injection of nifedipine did not significantly attenuate any of these hypertrophic responses in SHR. The activity of calcineurin in the heart was strongly suppressed by continuous but not bolus infusion of nifedipine in SHR. The results indicate that continuous blockade of Ca2+ channels with nifedipine effectively suppresses the development of cardiac hypertrophy in SHR, possibly through inhibition of the calcineurin activity. PMID- 11924718 TI - Expression and role of angiotensin II type 2 receptor in the kidney and mesangial cells of spontaneously hypertensive rats. AB - Angiotensin II type 2 (AT2) receptor is developmentally regulated and exerts antiproliferative and proapoptotic actions. Genetic ablation of this receptor in mice affects regulation of blood pressure, but the involvement of the AT2 receptor in the pathogenesis of hypertension remains unknown. In the present study, we examined developmental changes of angiotensin receptor subtypes in the kidney of stroke-prone spontaneously hypertensive rats (SHRSP), and compared them with those in normotensive Wistar-Kyoto rats (WKY). We also investigated the regulation and functional role of the AT2 receptor in cultured mesangial cells. Receptor binding and Northern blot analyses revealed that AT2 receptor expression is significantly lower in the SHRSP kidney than in the WKY kidney during the perinatal period, while AT1 receptor expression is not different between them. In WKY mesangial cells, AT2 receptor stimulation exerted a potent antiproliferative effect; this effect was not observed in SHRSP cells lacking the AT2 receptor expression. The expression of interferon regulatory factor (IRF)-1 paralleled the growth-dependent induction of AT2 receptor in WKY mesangial cells, and transfection of IRF-1 antisense oligonucleotide significantly suppressed AT2 receptor expression, indicating IRF-1-dependent regulation of AT2 receptor expression in mesangial cells. However, this induction was inefficient in SHRSP cells. Thus, we found impaired AT2 receptor expression in the SHRSP kidney in vivo and in mesangial cells in vitro. The unbalanced expression of renal angiotensin receptor subtypes with exaggerated AT1 receptor signaling during early life in SHRSP may play a role in the programming for hypertension and related renal injury. PMID- 11924719 TI - Kynureninase is a novel candidate gene for hypertension in spontaneously hypertensive rats. AB - The rostral ventrolateral medulla (RVLM) plays a critical role in the tonic and reflexive regulation of arterial blood pressure. Recent studies have demonstrated that injection of kynurenic acid (KYN) into the RVLM of spontaneously hypertensive rats (SHR) decreases arterial blood pressure. We hypothesized that a relative increase in the excitatory amino acid-mediated drive of RVLM vasomotor neurons in SHR may be due to derangement of one of the enzymes that affect the KYN level in the brain. We selected kynureninase, kynureninase hydroxylase, kynurenine aminotransferase type I, and kynurenine aminotransferase type II as candidates that may affect the KYN level in the brainstem. We conducted association studies between polymorphisms of these genes and blood pressure in an F2 population derived from SHR and Wistar-Kyoto rats (WKY). The cosegregation analysis indicated that only the kynureninase gene (KYNU) polymorphism influenced systolic blood pressure (SBP) and residuals of systolic blood pressure after adjusting for heart rate and body weight (RSBP). KYNU was found to be located on rat chromosome 3, and quantitative trait loci analysis at this locus indicated that the logarithms of the odds scores for KYNU in terms of SBP and RSBP were 2.0 and 3.3, respectively. This association with blood pressure decreased in proportion to the distance from KYNU. The expression level of KYNU mRNA in the brainstem was about 3.1 and 2.9 times higher in 10-week-old and 16-week-old SHR than in age-matched WKY, respectively. The increased expression of KYNU in SHR is thought to decrease the KYN level. KYNU seems to be one of the genes that contributes to hypertension in SHR. PMID- 11924720 TI - A case of metastatic extra-adrenal pheochromocytoma 12 years after surgery. AB - At the age of 53, a 65-year-old man had been diagnosed with extra-adrenal pheochromocytoma in the retroperitoneum and underwent total tumorectomy. Afterward, he had his serum catecholamine periodically measured in an outpatient clinic. In February 1999, 12 years after surgery, he complained of lower left abdominal pain. Computed tomography and magnetic resonance imaging revealed an osteolytic lesion in thoracic vertebrae 11Th (Th 11). Although his basal serum and urine catecholamines were at normal levels, glucagon injection increased blood pressure and plasma catecholamine levels. 131I-metaiodobenzylguanidine (MIBG) scintigraphy was specifically taken up to Th 11. By bone biopsy, the osteolytic lesion in Th 11 was finally diagnosed with metastasis of pheochromocytoma. For post-operative pheochromocytoma, long-term follow-up involving biochemical tests, including serum catecholamines, and MIBG is needed. PMID- 11924721 TI - Estimation of myocardial cell damage on the basis of mean electrocardiographic voltage and anatomical left ventricular mass. AB - Left ventricular mass (LVM) as assessed by magnetic resonance imaging (MRI, LVM(MRI)) and electrocardiographic (ECG) voltage reflect different pathological features. We hypothesized that ECG voltage is related to the electrical potential of cardiac muscle cells (electrical LVM) and to anatomical LVM as evaluated by MRI, and that the divergence between electrical LVM and anatomical LVM reflects the degree of myocardial damage. Because adipose tissue has high electrical resistance, we previously found a very strong correlation between body-fat corrected mean ECG voltage (Vfm) and LVM as estimated by echocardiography in patients with essential hypertension. In this study we compared LVM(MRI), Vfm, the ratio of Vfm x 10(2)/LVM(MRI), and the results of 99mTc tetrofosmin scintigraphy in patients with and without myocardial infarction (MI). We studied 33 patients without Ml and 26 patients with Ml. Vfm significantly correlated with LVM(MRI) in patients without MI (r=0.71, p<0.01). The ratio of Vfm x 10(2)/LVM(MRI) apparently reflected the relation between electrical LVM and anatomical LVM. Vfm x 10(2)/LVM(MRI) in patients with MI was smaller than that in patients without MI (0.98+/-0.28 vs. 1.42+/-0.29, p<0.01). Vfm x 10(2)/LVM(MRI) decreased as 99mTc score increased (r=-0.66, p<0.01). Our results indicate that Vfm is a useful index of electrical LVM and that Vfm x 10(2)LVM(MRI) reflects the electrical potential of the viable myocardium in total anatomical LVM. PMID- 11924722 TI - PPARgamma2 pro12Ala polymorphism and insulin resistance in Japanese hypertensive patients. AB - We investigated the relationship between peroxisome proliferator-activated receptor gamma (PPARgamma) Pro12Ala substitution and insulin resistance in subjects with normal insulin secretory capacity, since it has been reported that PPARgamma may affect not only insulin resistance but also insulin secretion. We examined 81 Japanese male patients with untreated essential hypertension using the glucose clamp technique. We found 77 subjects with Pro/Pro and 4 subjects with Pro/Ala genotype, and the glucose disposal rate was not significantly different between the two groups. Fasting plasma glucose, fasting immunoreactive insulin, total cholesterol, HDL cholesterol, and triglyceride were not significantly different between the two groups. There were also no significant differences between groups in homeostasis model assessment of insulin resistance (HOMA-R) values, area under the curve (AUC) for plasma glucose, or AUC for IRI in 75 g OGTT. Because insulin sensitivity is likely to be determined by polygenic factors, we also investigated beta3 adrenergic receptor Trp64Arg polymorphism as a possible determinant of insulin resistance. In conclusion, no significant association was observed between PPARgamma2 substitution and insulin sensitivity in the present cohort of Japanese hypertensive patients. PMID- 11924723 TI - Genotype-specific association between circulating soluble cellular adhesion molecules and carotid intima-media thickness in community residents: J-SHIPP study. Shimanami Health Promoting Program. AB - Plasma levels of soluble forms of cellular adhesion molecules (CAMs) and their relationships with carotid intima-media thickness (IMT) were investigated in community residents. Plasma levels of soluble intercellular adhesion molecule-1 (sICAM-1) and soluble vascular cell adhesion molecule-1 (sVCAM-1) were measured by ELISA in 200 community residents in Japan. Carotid IMT showed a weak but significant positive correlation with the plasma levels of both sICAM-1 (r=0.175, p=0.013) and sVCAM-1 (r=0.19, p=0.0075). Gene polymorphisms of angiotensin converting enzyme (ACE) insertion/deletion (I/D), angiotensinogen (AGT) M235T, angiotensin II type 1 receptor (AT1R) A1166C and apolipoprotein E (apoE) were determined for each subject. The plasma level of sVCAM-1 tended to be lower in subjects with the ACE DD genotype than in those with the ACE ID and II genotypes (373+/-94, 421+/-133, 443+/-135 ng/ml, respectively, p=0.056). However, there were no genotype-specific differences in the plasma levels of soluble forms of CAMs for the other genes examined. In a separate analysis, the plasma level of sICAM-1 was significantly associated with carotid IMT in ACE D carriers (ID + DD) (r=0.28, p=0.002), AGT M carriers (MT + MM) (r=0.32, p=0.0045), and subjects with apoE4 (r=0.35, p=0.036). In contrast, the plasma level of sVCAM-1 showed significant positive correlations with carotid IMT in subjects with the ACE II genotype (r=0.33, p=0.0027) or AGT TT genotype (r= 0.22, p=0.015), and subjects with apoE E2/E3 or E3/E3 (r=0.16, p=0.043). Stepwise regression analysis showed that plasma sVCAM-1 was independently associated with carotid IMT in subjects with the ACE II genotype or apoE4 genotype. Similarly, the plasma level of sICAM 1 was independently associated with carotid IMT in AGT M carriers. These findings suggest that genetic background could be involved in the association between plasma CAMs and atherosclerosis. PMID- 11924725 TI - Influence of plasma aldosterone on left ventricular geometry and diastolic function in treated essential hypertension. AB - Since aldosterone is known to promote interstitial fibrosis in cardiac tissues, it is possible that aldosterone may influence cardiac structure and function. In the present study, we investigated whether plasma aldosterone concentration (PAC) is related to the distinct patterns of left ventricular (LV) geometry and LV diastolic function in treated essential hypertension. In 92 patients with chronically treated essential hypertension, two-dimensional and Doppler echocardiographic examinations were performed and LV inflow velocities were measured for evaluation of LV diastolic function. When patients were divided into four groups by the different LV geometric patterns, PAC in patients with eccentric hypertrophy was significantly higher than in those with concentric hypertrophy (15.2+/-2.1 vs. 10.0+/-0.7 ng/dl, p<0.01). However, the ratio of the peak velocity of early diastolic filling to that of atrial filling (EIA), an index of LV diastolic function, was significantly decreased in patients with concentric hypertrophy compared with those showing normal geometry. In the relationship between PAC and LV diastolic function, PAC was negatively correlated with EIA (r=-0.35, p<0.05) only in the subgroup with normal relative wall thickness (i.e., without the concentric change in LV geometry). A multiple linear regression analysis showed that PAC was one of the independent determinants of E/A in the overall subject group. These observations indicate that PAC is associated with the eccentric change in LV geometry in patients with treated essential hypertension and also suggest that the increase in PAC participates in the impairment of LV diastolic function apart from the concentric change in LV geometry, although concentric hypertrophy clearly impairs LV diastolic function. PMID- 11924724 TI - Intensive blood pressure reduction is beneficial in patients with impaired cardiac function coexisting with chronic renal insufficiency. AB - Both in CHF (congestive heart failure) and CRI (chronic renal insufficiency), blood pressure reduction is beneficial for preservation of cardiac and renal function. However, it is uncertain how much blood pressure reduction is appropriate in patients with both CHF and coexisting CRI. In the present study, we examined whether intensive blood pressure reduction is more beneficial in these patients than the usually accepted level of reduction. Thirty-five men and 21 women of average age 63+/-5 years suffering from both CHF and CRI were selected from 316 patients attending the Kidney Disease Center of Saitama Medical School Hospital. All participants had an ejection fraction (EF) of less than 55% as determined by echocardiography. Renal function was evaluated by 24-h creatinine clearance (GFR), and a GFR of less than 50 ml/min was regarded as indicating renal insufficiency. Patients were divided into 2 groups according to the target blood pressure: in group I, blood pressure (BP) was lowered to less than 120/75 mmHg and in group II, blood pressure was lowered to less than 130/80 but more than 121/76 mmHg. The daily doses of basic antihypertensive agents were amlodipine 5 to 20 mg, benazepril 2.5 to 5 mg, guanabenz 2 to 8 mg and furosemide 20 to 60 mg. At the end of a 2-year follow-up period, the BP in group I was controlled at the level of 118+/-4/73+/-3 mmHg with good maintenance of EF (46+/ 4 to 60+/-4%) and GFR (44+/-4 to 40+/-3 ml/min). In group II, BP was maintained at 128+/-4/81+/-2 mmHg, accompanied by a reduction of EF (46+/-4 to 42+/-3%) and a significant reduction of GFR (44+/-3 to 35+/-3 ml/min). These results suggest that intensive blood pressure reduction might be beneficial in cases complicated by cardiorenal failure. PMID- 11924726 TI - Low-dose doxazosin improved aortic stiffness and endothelial dysfunction as measured by noninvasive evaluation. AB - Evaluation of atherosclerosis is important in the treatment of hypertension. To evaluate the preventive effects of a small amount of alpha-blockade, arterial and endothelial dysfunction were measured by noninvasive tests, i.e., pulse wave velocity, acceleration plethysmography and strain-gauge plethysmography, in patients with essential hypertension. Fifteen patients (65+/-3 years old) with essential hypertension (WHO stage I or II) were analyzed in this study. We performed noninvasive evaluations to measure aortic stiffness and endothelial dysfunction, in addition to measuring blood pressure, cholesterol profile, and levels of cells adhesion molecules and nitric oxide before and 6 and 12 months after the start of doxazosin treatment (1.0 mg/day). Blood pressure and heart rate did not significantly change during treatment. The pulse wave velocity index was significantly reduced both at 6 (7.72+/-0.23 m/s; p<0.05) and 12 (7.34+/-0.26 m/s; p<0.05) months after the start of treatment compared to the pretreatment level that at baseline. There was also a significant improvement in b/a after 12 months (-0.46+/-0.04; p<0.05) and in d/a after 6 months (-0.38+/-0.03; p<0.05) and 12 months (-0.39+/-0.03; p=0.05) compared to the pretreatment values. Moreover, reactive hyperemia evaluated by strain-gauge plethysmography after 6 months (1.34+/-0.11; p<0.05) and 12 months (1.49+/-0.16; p<0.05) was significantly improved compared to that before treatment, and NOx was significantly increased after 12 months (89.7+/-15.7 micromol/l; p<0.005). These data suggest that a low dose of doxazosin may play an important role in improving arterial stiffness and endothelial dysfunction without changing cardiac hemodynamics. PMID- 11924727 TI - Blood pressure control assessed by home, ambulatory and conventional blood pressure measurements in the Japanese general population: the Ohasama study. AB - To assess blood pressure control in the Japanese population, we analyzed previously obtained measurements of conventional, home and ambulatory blood pressures in 1,174 subjects aged > or =40 in a Japanese community. On the basis of conventional blood pressure values and the use of antihypertensive medication, participants were classified as normotensive, untreated hypertensive and treated hypertensive subjects. When 140/90, 135/85 and 135/85 mmHg were used as the hypertension criteria for conventional, home and ambulatory blood pressure measurements, respectively, all three blood pressure values were higher in untreated and treated hypertensive subjects than in normotensive subjects. Among the treated hypertensive subjects, approximately half were classified as hypertensive not only by conventional blood pressure, but also by home or ambulatory measurements. Approximately 10% of the subjects defined as normotensive by conventional blood pressure measurement were classified as hypertensive by home or ambulatory measurements, whereas 60% of the untreated hypertensive subjects as defined by conventional blood pressure measurement had normal home or ambulatory blood pressure values. Therefore, we concluded that 1) the poor blood pressure control in treated hypertensive subjects was attributable not only to the white coat effect but also to inadequate control of blood pressure; and 2) a certain percentage of subjects were misclassified as hypertensive or normotensive by conventional blood pressure measurement. PMID- 11924728 TI - Association study between a novel single nucleotide polymorphism of the promoter region of the prostacyclin synthase gene and essential hypertension. AB - The purpose of this study was to investigate whether an association exists between the promoter region of the prostacyclin synthase gene and essential hypertension (EH). Using the polymerase chain reaction-single strand conformation polymorphism (PCR-SSCP) method, we discovered a novel single nucleotide polymorphism (SNP), T-192G, in the 5'-flanking region. We performed an association study using the SNP in 200 patients and 200 controls. The allele frequency distribution in the two groups was not significantly different. Thus, this SNP in the PGIS gene is not associated with EH. PMID- 11924729 TI - Questionnaire survey on the Japanese guidelines for treatment of hypertension in the elderly: 1999 revised version. AB - A questionnaire survey was administered to Japanese clinical specialists in hypertension in order to gauge their opinions on the 1999 revised version of the Guidelines for Hypertension in the Elderly prepared by the Comprehensive Research Project on Aging and Health of the Ministry of Health and Welfare. Out of 162 council members of the Japanese Society of Hypertension, 122 (75%) replied. The majority (93%) of respondents approved of the guidelines in general, and 72% of them approved of the age-related setting of a therapeutic goal for blood pressure. Sixty-five percent of respondents selected long-acting Ca antagonists, ACE inhibitors and low-dose diuretics as first-line agents for hypertension without complications in the elderly. The results of the questionnaire survey should be reflected in the next version of the guidelines. PMID- 11924730 TI - A new method for evaluation of split renal cortical blood flow with contrast echography. AB - The recent development of contrast echography has made renal enhancement possible through an intravenous injection of microbubble-based contrast. In animal models, tissue perfusion can be quantified using contrast echography by measurement of the rate at which microbubbles replenish tissue after their ultrasound-induced destruction. Our purpose in this study was to evaluate renal blood flow with contrast echography in humans. To increase the sensitivity for microbubbles, we used a combination of power Doppler harmonic and intermittent imaging. The pulsing interval (PI) was changed from 10 cardiac cycles to 1 cardiac cycle during an intravenous infusion of the contrast agent, and alterations in the intensity of the renal cortex were represented as a decline ratio (DR). In 24 patients with various renal diseases, we were able to observe all 48 kidneys with adequate enhancement of the renal cortex. At PI of 10 cardiac cycles, the enhancement was homogeneous and strong, while, obviously, changing PI from 10 to 1 cardiac cycles caused a decline of enhancement. An excellent correlation was found between DR using contrast echography and renal plasma flow determined by clearance and radionuclide measurements. An excellent correlation was found between the DR values determined by contrast echography and the renal plasma flow values determined using clearance and radionuclide measurements. These results suggest that DR may be useful for evaluation of both total and split renal blood flow. Thus the contrast echographic method presented here could succeed in assessing renal cortical blood flow less invasively than conventional methods in humans. PMID- 11924731 TI - Perindopril reverses myocyte remodeling in the hypertensive heart. AB - Studies have shown that the renin-angiotensin system (RAS) plays an important role in cardiac remodeling induced by hypertension. However, the role of this system on myocyte remodeling remains unclear. In the present study, we have assessed the effect of perindopril, an angiotensin converting enzyme (ACE) inhibitor, in spontaneously hypertensive rats (SHRs) as a means to evaluate the role of RAS in myocyte remodeling. We also investigated the effect of beta blockade on myocyte remodeling. We used female SHRs at 12 weeks of age. They were divided into four experimental groups: a control group, group C; low dose perindopril group (0.3 mg/kg/day, p.o.), group PL; high dose perindopril group (3 mg/kg/day, p.o.), group PH; and bisoprolol group (60 mg/kg/day, p.o.), group B. We isolated myocytes from these rats after 4 weeks. LV myocyte volume and cross sectional area decreased in groups PL and PH compared to group C. LV myocyte length decreased in group PH compared to group C. However, there was no morphological change in LV myocytes in group B compared to group C. In summary, ACE inhibitors reversed cardiac hypertrophy mainly by a reduction in LV myocyte volume; however, beta blockade did not reverse myocyte remodeling. These results suggest that RAS plays an important role in myocyte remodeling in the hypertensive heart. PMID- 11924732 TI - A role of oxidative stress-generated eicosanoid in the progression of arteriosclerosis in type 2 diabetes mellitus model rats. AB - Diabetes mellitus (DM) is a well-established risk factor of cardiovascular diseases. We investigated the mechanism of the progression of arteriosclerosis in DM, focusing on the role of oxidative stress and insulin resistance in vivo. Male Otsuka Long-Evans Tokushima Fatty (OLETF) rats, an experimental model of type 2 DM, were assigned to 3 groups, based on supplementation with vitamin E (VE) or troglitazone (TR), a VE-derived agent which improves insulin-resistance. At 36 weeks, plasma and aortic tissue 8-iso-PGF2alpha contents, a vascular proliferating eicosanoid produced in vivo by oxidative stress, were measured by EIA. TGF-beta1 and TGF-beta1 receptor II were immunohistochemically analyzed. Histopathologically, medial area and the nuclear number of smooth muscle cells of the aorta were measured. The tissue 8-iso-PGF2alpha content (pg/g tissue) was significantly decreased by either VE or TR in the aorta (untreated-OLETF, 15,332+/-3,254 vs. TR-treated-OLETF, 7,092+/-1,992 or VE-treated-OLETF, 5,394+/ 836, both p<0.01), but that in plasma decreased by only VE. VE and TR improved the increased the level of the actual medial area and the number of smooth muscle cells. The expression of TGF-beta1 was reduced, but TGF-beta1 receptor II was not. 8-iso-PGF2alpha may play an important role in the progression of arteriosclerosis. Antioxidant treatment may promise significant clinical benefits in the early diabetic stage. PMID- 11924733 TI - Green coffee bean extract and its metabolites have a hypotensive effect in spontaneously hypertensive rats. AB - The effects of a water-soluble green coffee bean extract (GCE) on blood pressure were investigated using spontaneously hypertensive rats (SHR). There was a dose dependent reduction in blood pressure after a single ingestion (180 to 720 mg/kg, p.o.) or long-term ingestion (0.25 to 1% diet for 6 weeks) of GCE. A single oral ingestion (50 to 200 mg/kg) of 5-caffeoylquinic acid (5-CQA), the major component of GCE, dose-dependently decreased blood pressure, suggesting that 5-CQA is involved in the hypotensive effect of GCE in SHR. Because significant increases in caffeic acid (CA) or ferulic acid (FA) were detected in plasma after oral ingestion of 5-CQA in SHR, these acids (2.5, 5,10 micromol/kg) were intravenously injected into SHR under anesthesia and the carotid arterial pressure was measured. Of the two components, FA had a stronger depressor effect than CA. The depressor effect of FA (50 mg/kg, p.o.) was attenuated by the concurrent injection of atropine sulfate (5 mg/kg, s.c.), suggesting that the hypotensive effect of FA in SHR might be mediated via the muscarinic acetylcholine receptors. These findings indicate that oral ingestion of GCE or 5-CQA decreases blood pressure in SHR, and that FA, which is a metabolite of 5-CQA, is a candidate hypotensive component. PMID- 11924734 TI - Combined ab initio/empirical approach for optimization of Lennard-Jones parameters for polar-neutral compounds. AB - The study of small functionalized organic molecules in aqueous solution is a useful step toward gaining a basic understanding of the behavior of biomolecular systems in their native aqueous environment. Interest in studying amines and fluorine-substituted compounds has risen from their intrinsic physicochemical properties and their prevalence in biological and pharmaceutical compounds. In the present study, a previously developed approach which optimizes Lennard-Jones (LJ) parameters via the use of rare gas atoms combined with the reproduction of experimental condensed phase properties was extended to polar-neutral compounds. Compounds studied included four amines (ammonia, methylamine, dimethylamine, and trimethylamine) and three fluoroethanes (1-fluoroethane, 1,1-difluoroethane, and 1,1,1-trifluoroethane). The resulting force field yielded heats of vaporization and molecular volumes in excellent agreement with the experiment, with average differences less than 1%. The current amine CHARMM parameters successfully reproduced experimental aqueous solvation data where methylamine is more hydrophilic than ammonia, with hydrophobicity increasing with additional methylation on the nitrogen. For both the amines and fluoroethanes the parabolic relationship of the extent of methylation or fluorination, respectively, to the heats of vaporization were reproduced by the new parameters. The present results are also discussed with respect to the impact of parameterization approach to molecular details obtained from computer simulations and to the unique biological properties of fluorine in pharmaceutical compounds. PMID- 11924735 TI - Application of the frozen atom approximation to the GB/SA continuum model for solvation free energy. AB - The generalized Born/surface area (GB/SA) continuum model for solvation free energy is a fast and accurate alternative to using discrete water molecules in molecular simulations of solvated systems. However, computational studies of large solvated molecular systems such as enzyme-ligand complexes can still be computationally expensive even with continuum solvation methods simply because of the large number of atoms in the solute molecules. Because in such systems often only a relatively small portion of the system such as the ligand binding site is under study, it becomes less attractive to calculate energies and derivatives for all atoms in the system. To curtail computation while still maintaining high energetic accuracy, atoms distant from the site of interest are often frozen; that is, their coordinates are made invariant. Such frozen atoms do not require energetic and derivative updates during the course of a simulation. Herein we describe methodology and results for applying the frozen atom approach to both the generalized Born (GB) and the solvent accessible surface area (SASA) parts of the GB/SA continuum model for solvation free energy. For strictly pairwise energetic terms, such as the Coulombic and van-der-Waals energies, contributions from pairs of frozen atoms can be ignored. This leaves energetic differences unaffected for conformations that vary only in the positions of nonfrozen atoms. Due to the nonlocal nature of the GB analytical form, however, excluding such pairs from a GB calculation leads to unacceptable inaccuracies. To apply a frozen atom scheme to GB calculations, a buffer region within the frozen-atom zone is generated based on a user-definable cutoff distance from the nonfrozen atoms. Certain pairwise interactions between frozen atoms in the buffer region are retained in the GB computation. This allows high accuracy in conformational GB comparisons to be maintained while achieving significant savings in computational time compared to the full (nonfrozen) calculation. A similar approach for using a buffer region of frozen atoms is taken for the SASA calculation. The SASA calculation is local in nature, and thus exact SASA energies are maintained. With a buffer region of 8 A for the frozen-atom cases, excellent agreement in differences in energies for three different conformations of cytochrome P450 with a bound camphor ligand are obtained with respect to the nonfrozen cases. For various minimization protocols, simulations run 2 to 10.5 times faster and memory usage is reduced by a factor of 1.5 to 5. Application of the frozen atom method for GB/SA calculations thus can render computationally tractable biologically and medically important simulations such as those used to study ligand-receptor binding conformations and energies in a solvated environment. PMID- 11924736 TI - Linear scaling local correlation approach for solving the coupled cluster equations of large systems. AB - A linear scaling local correlation approach is proposed for approximately solving the coupled cluster doubles (CCD) equations of large systems in a basis of orthogonal localized molecular orbitals (LMOs). By restricting double excitations from spatially close occupied LMOs into their associated virtual LMOs, the number of significant excitation amplitudes scales only linearly with molecular size in large molecules. Significant amplitudes are obtained to a very good approximation by solving the CCD equations of various subsystems, each of which is made up of a cluster associated with the orbital indices of a subset of significant amplitudes and the local environmental domain of the cluster. The combined effect of these two approximations leads to a linear scaling algorithm for large systems. By using typical thresholds, which are designed to target an energy accuracy, our numerical calculations for a wide range of molecules using the 6-31G or 6-31G* basis set demonstrate that the present local correlation approach recovers more than 98.5% of the conventional CCD correlation energy. PMID- 11924737 TI - Ab initio prediction of helical segments in polypeptides. AB - An ab initio method has been developed to predict helix formation for polypeptides. The approach relies on the systematic analysis of overlapping oligopeptides to determine the helical propensity for individual residues. Detailed atomistic level modeling, including entropic contributions, and solvation/ionization energies calculated through the solution of the Poisson Boltzmann equation, is utilized. The calculation of probabilities for helix formation is based on the generation of ensembles of low energy conformers. The approach, which is easily amenable to parallelization, is shown to perform very well for several benchmark polypeptide systems, including the bovine pancreatic trypsin inhibitor, the immunoglobulin binding domain of protein G, the chymotrypsin inhibitor 2, the R69 N-terminal domain of phage 434 repressor, and the wheat germ agglutinin. PMID- 11924738 TI - Support Vector Machines for predicting HIV protease cleavage sites in protein. AB - Knowledge of the polyprotein cleavage sites by HIV protease will refine our understanding of its specificity, and the information thus acquired is useful for designing specific and efficient HIV protease inhibitors. The pace in searching for the proper inhibitors of HIV protease will be greatly expedited if one can find an accurate, robust, and rapid method for predicting the cleavage sites in proteins by HIV protease. In this article, a Support Vector Machine is applied to predict the cleavability of oligopeptides by proteases with multiple and extended specificity subsites. We selected HIV-1 protease as the subject of the study. Two hundred ninety-nine oligopeptides were chosen for the training set, while the other 63 oligopeptides were taken as a test set. Because of its high rate of self consistency (299/299 = 100%), a good result in the jackknife test (286/299 = 95%) and correct prediction rate (55/63 = 87%), it is expected that the Support Vector Machine method can be referred to as a useful assistant technique for finding effective inhibitors of HIV protease, which is one of the targets in designing potential drugs against AIDS. The principle of the Support Vector Machine method can also be applied to analyzing the specificity of other multisubsite enzymes. PMID- 11924739 TI - Prediction of aqueous solubility of drugs and pesticides with COSMO-RS. AB - The COSMO-RS method, originally developed for the prediction of liquid-liquid and liquid-vapor equilibrium constants based on quantum chemical calculations, has been extended to solid compounds by addition of a heuristic expression for the Gibbs free energy of fusion. By this addition, COSMO-RS is now capable of a priori prediction of aqueous solubilities of a wide range of typical neutral drug and pesticide compounds. Only three parameters in the heuristic expression have been fitted on a data set of 150 drug-like compounds. On these data an rms deviation of 0.66 log-units was achieved. Later, the model was tested on a set of 107 pesticides, which have been critically selected based on two experimental data sources and by a crosscheck with an independent HQSAR model. On this data set an rms of 0.61 log-units was achieved, without any adjustments to the structurally extremely diverse pesticides. This result verifies the ability of this extended COSMO-RS to predict aqueous solubilities of drugs and pesticides of almost arbitrary structural classes. The new method is COSMO-RSol. PMID- 11924740 TI - MM3 parametrization of four- and five-coordinated rhenium complexes by a genetic algorithm--which factors influence the optimization performance? AB - Genetic algorithms (GA) were used to solve one of the multidimensional problems in computational chemistry, the optimization of force field parameters. The correlation between the composition of the GA, its parameters (p(c), p(m)) and the quality of the results were investigated. The composition was studied for all combinations of a Simple GA/Steady State GA with a Roulette Wheel/Tournament Selector using different values each for crossover (0.5, 0.7, 0.9) and mutation rates (0.01, 0.02, 0.05, 0.10, 0.20). The results show that the performance is strongly dependent on the GA scheme, where the Simple GA/Tournament Selector yields the best results. Two new MM3 parameters were introduced for rhenium compounds with coordination number four (204) and coordination number five (205), the formal oxidation states of rhenium ranging from +V to +VII. A manifold of parameters (Re-C, N, O, S) was obtained by using a diverse set of CSD structures. The advantage of the GA vs. UFF calculations is shown by comparison of several examples. The GA optimized parameters were able to reproduce the geometrical data of the X-ray structures. PMID- 11924741 TI - Mechanism for large first hyperpolarizabilities of phosphonic acid stilbene derivatives. AB - This paper presents calculations of dipole moments (mu), static polarizabilities (alpha), and first hyperpolarizabilities (beta) of phosphonic acid stilbene derivatives calculated in the framework of density functional theory. These calculations were performed using a finite field approach implemented in the density functional program ALLCHEM and were of an all-electron type using local exchange-correlation functional and specially designed basis sets. The molecular structures have been fully optimized using the semiempirical program MSINDO. Some of the investigated stilbenes have been synthesized very recently while others are described for the first time. Donor and acceptor groups of these analogues have been modified and the influence of these changes on the first hyperpolarizabilities has been investigated. This work demonstrates that the nonlinear optical response beta of these compounds increases dramatically when the acceptor moiety is displaced by analogues containing alkali metal groups. A general mechanism for the design of novel nonlinear optical materials with large first hyperpolarizabilities is described. PMID- 11924742 TI - Chemical shift driven geometry optimization. AB - A new method for refinement of 3D molecular structures by geometry optimization is presented. Prerequisites are a force field and a very fast procedure for the calculation of chemical shifts in every step of optimization. To the energy, provided by the force field (COSMOS force field), a pseudoenergy, depending on the difference between experimental and calculated chemical shifts, is added. In addition to the energy gradients, pseudoforces are computed. This requires the derivatives of the chemical shifts with respect to the coordinates. The pseudoforces are analytically derived from the integral expressions of the bond polarization theory. Single chemical shift values attributed to corresponding atoms are considered for structural correction. As a first example, this method is applied for proton position refinement of the D-mannitol X-ray structure. A crystal structure refinement with 13C chemical shift pseudoforces is carried out. PMID- 11924743 TI - Efficient linear algebra routines for symmetric matrices stored in packed form. AB - Quantum chemistry methods require various linear algebra routines for symmetric matrices, for example, diagonalization or Cholesky decomposition for positive matrices. We present a small set of these basic routines that are efficient and minimize memory requirements. PMID- 11924744 TI - Spectral density calculation by using the Chebyshev expansion. AB - A method to calculate the spectral density of any state vectors with respect to a set of eigenstates of a Hamiltonian is presented. A spectral density operator, whose expectation value on the state vector gives the spectral density, is evaluated indirectly by using the Chebyshev expansion method. A spectral transformation function is introduced to improve resolution at the low energy region, at the expense of the one at the higher region. The predissociation spectrum of CO+ is calculated to demonstrate the method. PMID- 11924745 TI - Neuroticism and low self-esteem as risk factors for psychosis. AB - BACKGROUND: Low self-esteem and high neuroticism are common features in psychosis, but in the absence of longitudinal studies it is unclear whether they represent consequences of the illness or risk factors acting before illness onset. METHODS: A population sample of 3,929 individuals with no lifetime evidence of psychosis were interviewed with the Composite International Diagnostic Interview and were administered the Groningen Neuroticism Scale and the Rosenberg Self-Esteem Scale at baseline and 1 and 3 years later. At year 3, individuals with CIDI evidence of psychotic symptoms were interviewed by clinicians to identify incident psychotic or psychosis-like symptoms. RESULTS: Baseline neuroticism and self-esteem predicted first-ever onset of psychotic symptoms at year 3 (neuroticism, OR 1.16, 95% CI 1.09, 1.23; self-esteem, OR 1.09, 95% CI 1.01, 1.18).When adjusted for each other and for level of anxiety and depression, neuroticism was the strongest independent predictor for onset of psychotic symptoms (OR 1.16, 95% CI 1.07, 1.26). CONCLUSIONS: Neuroticism increases the risk for development of psychotic symptoms. Mechanisms of risk may involve certain cognitive styles associated with neuroticism, such as beliefs about the uncontrollability of certain events and experiences. The association between low self-esteem and psychosis may involve the area of overlap between self-esteem and neuroticism. PMID- 11924747 TI - Outcome measures of interepisode bipolar patients in a Turkish sample. AB - BACKGROUND: We aimed to study the correlations of several outcome measures in bipolar patients with the clinical features of interepisode period. METHODS: Bipolar patients who were diagnosed according to DSM-III-R or IV were contacted and asked for a further evaluation. Interepisode bipolar patients (n = 100) were interviewed with the Schedule for Affective Disorders and Schizophrenia (SADS). In addition the Brief Disability Questionnaire (BDQ), the Quality of Life Enjoyment and Satisfaction Questionnaire (Q-LES-Q) and the Global Assessment Scale (GAS) were administered to assess outcome. They were also asked to check the List of Life Events (LLE) for the last six months. RESULTS: Our results can be summarised as follows: (1) quality of life was predicted by current subthreshold depressive symptoms; (2) the number of previous depressive episodes, current subthreshold depressive and manic symptoms predicted disability; (3) the number of previous depressive episodes and the duration of hospitalisation as well as current subthreshold depressive and manic symptoms predicted overall functioning; (4) the number and distress level of life events were correlated with suicidal symptoms. CONCLUSIONS: Our findings suggest that outcome measures were correlated with subsyndromal disorder, the number of previous depressive episodes and the duration of hospitalisation. PMID- 11924746 TI - Health and well-being among 70-year-old migrants living in Sweden--results from the H 70 gerontological and geriatric population studies in Goteborg. AB - BACKGROUND: Comparative epidemiological studies in migrants in Sweden have shown increased prevalence of psychosocial morbidity in young adults, but there is paucity of information on health in people aged 65 years and over. AIMS: We aim to compare prevalence of mental, physical and social health problems, rates of hospital admission, and mortality in Swedish and non-Swedish born people aged 70 years living in Goteborg, Sweden, and examine associations between social factors, physical health, mood and life satisfaction in the ethnic groups. In addition, we aim to test for the hypothesis that differences in mental health between migrants and natives are explained by social disadvantages rather than ethnicity. METHOD: Semi-structured interviews were administered to 84 migrants (47 women and 37 men randomly selected) with the help of bilingual interpreters, and 409 Swedes (183 males and 226 females) used as "controls". Complementary health and social data obtained from official sources on the total sample (N = 764, including non-participants in overall medical interviews) were used in comparative analyses of in-patient care and mortality and to check for the possibility of sampling bias. RESULTS: Migrants--originating mainly from Estonia, Poland, Yugoslavia, Germany, Italy and Nordic countries other than Sweden- reported more dizziness, poor vision and urinary problems, and fewer gall bladder problems than indigenous people (p < 0.05). Migrants also had higher levels of anxiety and depression and bodily pain, and lower levels of general health, social and emotional functioning, satisfaction with physical health status, family contacts, housing conditions and economic status than natives (p <0.05). Satisfaction with physical health seemed to be one of the strongest factors related to a lower prevalence of anxiety and depression together with family support and time spent in leisure pursuits. No significant differences in inpatient care in several ICD categories and all-cause mortality were found between overall migrants and the control group notwithstanding differences in self-reported health. CONCLUSION: The results indicate poorer subjective health in older migrants than natives in Goteborg, and also point to a "healthy migrant" effect on survival. PMID- 11924748 TI - Causal explanations of distress and general practitioners' assessments of common mental disorder among punjabi and English attendees. AB - BACKGROUND: The literature on the primary care assessment of mental distress among Indian subcontinent origin patients suggests frequent presentations to general practitioner, but rarely for recognisable psychiatric disorders. This study investigates whether cultural variations in patients' causal explanatory models account for cultural variations in the assessment of non-psychotic mental disorders in primary care. METHODS: In a two-phase survey, 272 Punjabi and 269 English subjects were screened. The second phase was completed by 209 and 180 subjects, respectively. Causal explanatory models were elicited as explanations of two vignette scenarios. One of these emphasised a somatic presentation and the other anxiety symptoms. Psychiatric disorder was assessed by GPs on a Likert scale and by a psychiatrist on the Clinical Interview Schedule. RESULTS: Punjabis more commonly expressed medical/somatic and religious beliefs. General practitioners were more likely to assess any subject giving psychological explanations to vignette A and English subjects giving religious explanations to vignette B as having a significant psychiatric disorder. Where medical/somatic explanations of distress were most prevalent in response to the somatic vignette, psychological, religious and work explanations were less prevalent among Punjabis but not among English subjects. Causal explanations did not fully explain cultural differences in assessments. CONCLUSIONS: General practitioners' assessments and causal explanations are related and influenced by culture, but causal explanations do not fully explain cultural differences in assessments. PMID- 11924749 TI - Psychiatric disorders in adult children of parents with a history of psychopathology. AB - BACKGROUND: The relation between major categories of psychiatric problems in parents and psychiatric disorders in their adult children has been investigated in only a few community studies. METHODS: In this study, data from a representative sample of the Dutch population (N = 7147) (response rate: 69.7 %) were used to examine this relation. DSM-III-R disorders were assessed using the CIDI. Parental psychiatric symptoms and childhood adversities were assessed using self-report measures. RESULTS: It was found that psychiatric symptoms in parents are strongly related to psychiatric disorders in their (adult) children, independent of type of parental psychiatric symptom. Only maternal problem drinking was not significantly related to an increased risk for their children. When controlled for childhood adversities and demographic variables, most relations between psychiatric disorders and parental psychiatric symptoms remained significant. Parental anxiety symptoms, however, were not significantly related to psychiatric disorders in the children, including anxiety disorders. It was also found that anxiety disorders in the children were not related to most parental psychiatric symptoms. CONCLUSIONS: This study has once more made it clear that children whose parents have psychiatric problems constitute an important high-risk group and that prevention and early intervention in these children constitute an important public health issue. PMID- 11924750 TI - Pneumonia-associated acute glomerulonephritis. AB - OBJECTIVE: Post-infectious glomerulonephritis typically occurs 7-14days after an infection. However, in several children we observed acute glomerulonephritis (AGN) to develop concurrently with pneumonia. The objective of the study was to delineate the clinical course and outcome of pneumonia-associated AGN. STUDY DESIGN: The hospital database was searched from 1984 - 1999 for c+hildren admitted with both acute pneumonia and AGN, each diagnosis having been made within 72 hours of each other. RESULTS: 11 boys, age 3.8- 12.7 years, were identified. Ten children had lobar pneumonia and I had an interstitial infiltrate. All responded to antibiotic therapy with resolution of fever and respiratory symptoms. Only I child developed an empyema. The mean +/- SD hospital stay was 5.9 +/- 3.9 days. All patients had an abnormal urinalysis with hematuria (gross hematuria in 5), proteinuria and cellular casts. At presentation, 7 children had a serum creatinine > 1.0 mg/dl and creatinine clearance < or = 80 ml/min/1.73 m2; in all, serum creatinine returned to normal and the creatinine clearance was > 80 ml! min/1.73 m2 on follow-up. Nine of the 11 children had a low serum complement C3, 3 of whom also had low complement C4. Anti-streptolysin O (ASO) titers were elevated in all 10 children tested. Six children developed hypertension and received antihypertensive medications. Only I child was severely oliguric requiring peritoneal dialysis for 4 days. He underwent a kidney biopsy, which showed acute proliferative glomerulonephritis without crescents. Neither a biopsy nor dialysis was performed in the other children. At follow-up, blood pressure, urinalysis and serum complements had normalized in the 9 children in whom follow-up was available. CONCLUSION: Children with pneumonia who are found to have abnormal urinalysis. hypertension, azotemia or oliguria should be evaluated for concomitant glomerulonephritis. In most children, pneumonia associated AGN runs a benign course and has a good prognosis, however, in some short-term medical intervention may be necessary. PMID- 11924752 TI - Polycystic kidney disease at end-stage renal disease in the United States: patient characteristics and survival. AB - BACKGROUND: The patient characteristics and mortality associated with autosomal dominant polycystic kidney disease have not been characterized for a national sample of end-stage renal disease (ESRD) patients. METHODS: 375,152 patients in the United States Renal Data System were initiated on ESRD therapy (including patients who eventually received renal transplants) between January 1, 1992 and June 30, 1997 and analyzed in an historical cohort study of polycystic kidney disease. RESULTS: Of the study population, 5,799 (1.5%) had polycystic kidney disease. In logistic regression, polycystic kidney disease was associated with Caucasian race (odds ratio 3.31, 95% CI, 3.09-3.54), women (1.10, 1.04-1.16), receipt of renal transplant (4.15, 3.87-4.45), peritoneal dialysis (vs. hemodialysis, 1.37, 1.27-1.49), younger age, and more recent year of first treatment for ESRD. Use of pre-dialysis EPO but not the level of serum hemoglobin at initiation of ESRD was significantly higher in patients with polycystic kidney disease. Patients with polycystic kidney disease had lower mortality compared to patients with other causes of ESRD, but patients with polycystic kidney disease had a higher adjusted risk of mortality associated with hemodialysis (vs. peritoneal dialysis) compared to patients with other causes of ESRD (hazard ratio 1.40, 1.13-1.75). CONCLUSIONS: Hematocrit at presentation to ESRD was not significantly different in patients with polycystic kidney disease compared with patients with other causes of ESRD. Peritoneal dialysis is a more frequent modality than hemodialysis in patients with polycystic kidney disease, and patients with polycystic kidney disease had an adjusted survival benefit associated with peritoneal dialysis, compared to patients with other causes of renal disease. PMID- 11924751 TI - Dyspepsia and gastroparesis in chronic renal failure: the role of Helicobacter pylori. AB - AIMS: Many patients with chronic renal failure have dyspeptic symptoms. In the present study, we assessed the Helicobacter pylori (Hp) status, dyspeptic symptoms and gastric emptying rates in uremic patients. The present study was undertaken to compare chronic renal failure patients not under dialysis therapy (predialysis), hemodialysis (HD) patients and peritoneal dialysis (PD) patients for these variables and to search for a possible causative role of Hp. METHODS: We used a standardized questionnaire to assess dyspeptic symptoms. Gastric emptying rates were determined by the 13C-octanoic acid breath test. HD patients were examined outside a dialysis session, PD patients were examined with a "full" abdomen. Specific Helicobacter pylori IgG was measured by a second-generation enzyme-linked immunosorbent assay. RESULTS: Sixty-six HD patients. 58 predialytic patients and 28 PD patients were included. Prevalences of Hp infection were highest in HD patients (46.2%) and predialysis patients (42.3%) compared to PD patients (28.6%) (p < 0.02). On the contrary, the prevalence of dysmotility-like dyspepsia was higher in PD patients (67.9%) when compared to HD patients (33.3%) (p < 0.01) and predialytic patients (53.6%) (difference not significant). Neither dyspepsia nor delayed gastric emptying were related to the presence of Helicobacter pylori IgG antibodies. CONCLUSION: A positive Helicobacter status based on serology was not related to the presence of dyspepsia or gastroparesis in uremic patients, whether on dialysis therapy or not. Dyspeptic complaints as well as gastroparesis are most prevalent in patients on peritoneal dialysis. The physiopathological mechanisms and clinical impact of these findings merit further investigation. PMID- 11924753 TI - Accuracy and clinical utility of dialysis dose measurement using online ionic dialysance. AB - BACKGROUND: Statistical associations between urea removal and survival have been described in a number of publications. Urea removal during treatment may be quantified by the delivered dose of dialysis. Methods in clinical use to measure delivered dose are retrospective and reliant on accurate blood sampling. The new generation of single patient proportionating systems incorporate the facility to automatically measure ionic dialysance throughout dialysis. METHODS: In a prospective study on 9 anuric patients with a stable dialysis prescription, we have compared the agreement of the dose of dialysis determined from ionic dialysance (Dt/V) with that derived from equilibriated Kt/V (eKt/V) and Kt/V measured by direct dialysis quantification (Kt/V(DDQ)) using 2 types of hemodialysis membrane (hemophan and low-flux polysulfone). The variability of the delivered dose over a 1-month period was also determined. RESULTS: Ionic dialysance was independent of membrane type. It was comparable to that established for plasma urea water clearance for hemophan but lower for polysulfone (p < 0.001). The mean (+/- SD) delivered dose of dialysis (Dt/V) was similar for both membranes (1.18 +/- 0.15 (hemophan) and 1.18 +/- 0.11 (low-flux polysulfone)). Bland Altman comparisons showed the limits of agreement between Dt/V and Kt/V(DDQ) were +/- 0.17 and for Dt/V compared with eKt/V +/- 0.15. A 1 month measurement of Dt/V demonstrated considerable treatment to treatment variability indicating that delivered dose cannot be considered stable. CONCLUSION: The availability of online measurement of ionic dialysance provides a step towards monitoring dialysis more closely at the time of delivery, and its clinical application will ensure that a more constant dialysis dose is delivered. PMID- 11924755 TI - Ultrasound changes of the carpal tunnel in patients receiving long-term hemodialysis: a cross-sectional and longitudinal study. AB - BACKGROUND: Carpal tunnel syndrome (CTS) is one of the major problems of long term hemodialysis (HD), but sometimes difficult to distinguish from uremic or diabetic neuropathy by clinical symptoms. PATIENTS AND METHODS: To evaluate the diagnosis of CTS more precisely, we examined the ultrasonographic alterations of the carpal tunnel and tendons of 90 wrists from 45 patients undergoing HD for more than 5 years. We measured the thickness of the palmar radiocarpal ligament (PRL), corresponding to the posterior wall of the carpal tunnel (CT), and the width of the CT, and compared those values with sensory (SCV), motor conduction velocity (MCV) of the median nerve and clinical symptoms. In addition, we longitudinally measured CT and PRL in the same patients for 5 years, and compared ultrasonographic changes and clinical parameters. RESULTS: A linear positive relationship was found between HD duration and PRL thickness (r = 0.43, p < 0.01) or CT width (r = 0.53, p < 0.01). CT diameter was negatively correlated with MCV (r = -0.30, p < 0.01) and SCV (r = -0.33, p < 0.04). PRL thickness was also inversely correlated with MCV (r = -0.44, p < 0.01) and SCV (r = -0.46, p < 0.01) of the median nerve, respectively. The wrists with clinical CTS and/or previous CTS surgery had significantly greater CT and PRL values compared to patients without CTS (CT: 6.1 0.2vs. 8.0+/-0.3 mm,p<0.01;PRL: 1.9+/-0.1 vs. 3.6 +/- 0.2 mm, p < 0.01). There was a significant increase in CT width from 6.2 +/- 0.2 to 7.1 +/- 0.2 mm (p < 0.01) and PRL thickness from 2.4 +/- 0.2 to 2.8 +/- 0.2 mm (p <0.01) during the 5-year observation, respectively. PRL thickness was constantly increased at the rate of 0.4 mm during the study. However, no significant association was found between the 5-year increases in CT and PRL distance and age, gender, the prevalence of diabetes, or laboratory parameters such as blood beta2-microglobulin, pentosidine and Kt/V(urea). CONCLUSION: Our data suggest that echographic evaluation of the wrist tissue thickness was useful to assess the progression of CTS. Serial measurements of the wrist by echography may be helpful to clarify the advance of subclinical CTS in patients receiving long-term HD. PMID- 11924754 TI - Lymphopenia in dialysis patients: a preliminary study indicating a possible role of apoptosis. AB - Lymphopenia is a common finding in dialysis patients. Since infection rate and mortality associated with infection are high in dialysis patients, lymphopenia may be one of the contributing factors. In the present study, we evaluated the mechanism responsible for lymphopenia in these patients. Lymphocytes isolated from dialysis patients showed increased apoptosis (p < 0.001) when compared to lymphocytes isolated from healthy subjects (healthy subjects, 0.5 +/- 0.2% vs. dialysis patients, 8.8 +/- 0.7% apoptotic cells/field). Sera from dialysis patients promoted lymphocyte apoptosis in a time- and dose-dependent manner. These sera also enhanced lymphocyte DNA fragmentation into multiple integers of 180 base pairs in the form of a ladder pattern. Cellulose acetate membranes promoted T cell apoptosis when compared to polysulfone membranes and to control. Cellulose acetate dialysis membranes also appear to promote lymphocyte FasL expression. Similarly, dialysis sera enhanced T cell Fas as well as FasL expression. Neither the cellulose acetate nor polysulfone membranes could induce FasL expression on B cells. Similarly, dialysis sera failed to induce FasL expression on B cells. On the other hand, anti-FasL antibodies attenuated dialysis sera-induced apoptosis in T as well as B cells. Interestingly, dialysis serum showed a 5-fold increase in FasL content when compared with control serum. These results suggest that dialysis-associated factors can induce autocrine death in T cells but the help of activated T cells is required to induce death in B cells. PMID- 11924756 TI - Epoetin omega for treatment of anemia in maintenance hemodialysis patients. AB - After the synthesis of epoetins alpha and -beta, a third molecule of recombinant human erythropoietin (rHuEPO) was synthesized and was named epoetin-omega. The molecule of epoetin-omega is a sialoglycoprotein with smaller amounts of O-bound sugars, less acidic and with different hydrophylity than the other 2 epoetins. The purpose of the study was to assess the efficacy, safety and clinical tolerance of epoetin-omega for treatment of renal anemia. In an open-label, uncontrolled prospective clinical study, 22 end-stage renal disease patients (9 male and 13 female) were followed for 6 months. They all had a hemoglobin (Hb) value below 85 g/l, and were on regular hemodialysis therapy 3 times a week, 4 hours per session. The initial weekly dose of epoetin-omega was 90 units per kg of body weight (b.w.) divided in 3 equal portions and administered subcutaneously after each dialysis session. After correction of the hemoglobin, the dose of rHuEPO was individualized to keep Hb within target limits of 100-120 g/l. To follow efficacy and safety, a number of clinical and laboratory parameters were monitored. All patients responded well to the therapy with corrected hemoglobin after the 10th week of the study. The mean dose of epoetin-omega during the correction period never exceeded 100 U/kg b.w. per week. The average maintenance dose of rHuEPO was 50-60 U/kg b.w. per week. Iron was, where needed, supplied intravenously. We noted no change in serum urea. creatinine, phosphorus, and heparin dose per dialysis session. The prothrombin time improved during the study. Serum albumin increased. No change was observed in urea reduction ratio (URR), body weight and mean arterial pressure. One serious adverse event was noted: worsening of hypertension in 1 patient, with the development of hypertensive encephalopathy and severe headache. rHuEPO treatment was stopped. The blood pressure was effectively controlled by reducting her body weight by 5%. Thereafter, rHuEPO therapy was resumed with good blood pressure control. We could conclude that recombinant human erythropoietin-omega was an efficient and safe therapeutic agent for the treatment of renal anemia. PMID- 11924758 TI - Tuberous sclerosis complex diagnosed firstly in an elderly patient. PMID- 11924757 TI - Acute endocapillary proliferative glomerulonephritis associated with human parvovirus B19 infection. AB - A 36-year-old female developed acute nephritic syndrome associated with human parvovirus B19 (HPVB19) infection. Laboratory data showed proteinuria, hypocomplementemia, mild pancytopenia, the presence of immunoglobulin (Ig) M and IgG antibodies to HPVB 19 and positive reaction of serum HPVB19 DNA using a polymerase chain reaction (PCR). A renal biopsy showed endocapillary hypercellularity mainly of mononuclear cells with segmental apparent mesangiolytic change; fine granular IgM, IgG and C3 deposits were noted by immunofluorescence microscopy; relatively small electron-dense deposits were observed in the widened subendothelial spaces and the mesangium, and loosening of the mesangial matrix varied from place to place electron microscopically. PCR of HPVB19 DNA in the renal biopsy tissue was positive as well as in the peripheral blood. The histological findings suggested that immune-complex-mediated endocapillary proliferative glomerulonephritis is caused by acute HPVB 19 infection. We discuss the differences from poststreptococcal glomerulonephritis and the possible pathogenesis of acute endocapillary proliferative glomerulonephritis associated with HPVB19 infection. PMID- 11924759 TI - Radioprotection by a herbal preparation of Hippophae rhamnoides, RH-3, against whole body lethal irradiation in mice. AB - PURPOSE: Hippophae rhamnoides L. has been well documented to have anti-oxidative, immunostimulative and regenerative properties and therefore a herbal preparation of H. rhamnoides coded as RH-3 was investigated for its radioprotective action. MATERIALS AND METHODS: RH-3 was administered intraperitonially (i.p.) to mice 30 minutes before whole body irradiation and whole body survival, spleen Colony forming units (CFU) and haematological parameters were studied. To investigate free radical scavenging and antioxidant potential, Fenton reaction, radiation mediated OH radical scavenging and chemically generated superoxide anions scavenging were studied in vitro while inhibition of lipid peroxidation was studied in liver homogenate of mice. RESULTS: A dose of 30 mg/kg body weight of RH-3 rendered 82% survival as compared to no survival in irradiated control. The endogenous CFU counts in mouse spleen on 10th post-irradiation day with and without RH-3 demonstrated radioprotective effect. Various hematological parameters also corroborated the radioprotective effect of RH-3. In a dose dependent manner, RH-3 inhibited Fenton reaction and radiation mediated generation of hydroxyl radicals in vitro, superoxide anion mediated Nitroblue tetrazolium (NBT) reduction and FeSO4 mediated lipid peroxidation in liver. CONCLUSION: Free radical scavenging, acceleration of stem cell proliferation and immunostimulation are the radioprotective attributes, which require further investigations. PMID- 11924760 TI - Antimutagenic and anticarcinogenic effects of Phyllanthus amarus. AB - This study aimed to examine the antimutagenic and anticarcinogenic potential of Phyllanthus amarus Schum. et Thonn. using the bacterial preincubation mutation assay and an in-vivo alkaline elution method for DNA single-strand breaks in hamster liver cells. The aqueous extract of the entire plant showed an antimutagenic effect against induction by 2-aminofluorene (AF2), 2 aminoanthracene (2AA) and 4-nitroquinolone-1-oxide (4-NQO) in Salmonella typhimurium strains TA98 and TA100, and in Escherichia coli WP2 uvrA/pKM101. All the results were dose-dependent; however, inhibition of N-ethyl-N nitrosoguanidine (ENNG)-induced mutagenesis was observed only with S. typhimurium TA100. The extract also exhibited activity against 2-nitrofluorene (2NF) and sodium azide-induced mutagenesis with S. typhimurium TA98 and TA100, respectively. Based on the alkaline elution method, the plant extract prevented in vivo DNA single-strand breaks caused by dimethylnitrosamine (DMN) in hamster liver cells. When the extract was administered 30 min prior to the administration of DMN, the elution rate constant decreased more than 2.5 times, compared to that of control. These results indicate that P. amarus possesses antimutagenic and antigenotoxic properties. PMID- 11924761 TI - Recent findings of green tea extract AR25 (Exolise) and its activity for the treatment of obesity. AB - The green tea extract AR25 is an 80% ethanolic dry extract standardized at 25% catechins expressed as epigallocatechin gallate (EGCG). In vitro, green tea extract AR25 exerts a direct inhibition of gastric and pancreatic lipases and a stimulation of thermogenesis. In an open study, the effects of extract AR25 were evaluated in moderately obese patients. After 3 months, body weight was decreased by 4.6% and waist circumference by 4.48%. These results suggest the green tea extract AR25 to be a natural product for the treatment of obesity, which exerts its activity by several ways: inhibition of lipases and stimulation of thermogenesis. PMID- 11924762 TI - Comparison of antioxidative capacities and inhibitory effects on cholesterol biosynthesis of quercetin and potential metabolites. AB - The flavonol quercetin is known to be rapidly metabolized after ingestion by enterocytes and bacteria in the intestinal tract which may influence the biological, e.g. antioxidative potency of this compound. Therefore, quercetin and several of its possible metabolites were compared with regard to their antioxidant activity and their capacity to inhibit hepatocellular cholesterol biosynthesis. Using the 2,2,-diphenylpicrylhydrazyl radical scavenger assay, all compounds with an ortho diphenolic structure acted as strong antioxidants. In contrast, in a cellular assay focusing on lipid peroxidation in cultured rat hepatocytes challenged with tert.-butylhydroperoxide only the lipophilic compounds quercetin and 3,4-dihydroxytoluene were active. Concerning the inhibition of cholesterol biosynthesis, 3,4-dihydroxytoluene surprisingly mimicked the effect of quercetin in primary rat hepatocytes, but much less so in HepG2 cells. All other metabolites were almost ineffective in both cell types. These results suggest that some of the biological functions of flavonoids detectable by in vitro assays may persist in vivo as long as comparably potent metabolites are systemically present. PMID- 11924763 TI - Cyclooxygenase enzyme inhibitory compounds with antioxidant activities from Piper methysticum (kava kava) roots. AB - Cyclooxygenase enzyme inhibitory assay-guided purification of ethyl acetate extract of Piper methysticum (kava kava) roots yielded six biologically active compounds (1-7), which were purified using MPLC, preparative TLC and HPLC methods. These compounds were also evaluated for antioxidant activities. Dihydrokawain (1) and yangonin (6) showed the highest COX-I and COX-II inhibitory activities at 100 microg/ml, respectively. The lipid oxidation assay did not reveal antioxidant activities for demethoxyangonin (2), dihydrokawain (1), kawain (4), dihydromethysticin (5) or methysticin (7) at 50 microg/ml. The antioxidant activities of flavokawain A (3) and yangonin (6) could not be tested in the lipid oxidation assay due to solubility problems. However, yangonin and methysticin showed moderate antioxidant activities in the free radical scavenging assay at 2.5 mg/ml. PMID- 11924764 TI - Estrogenic and cholinergic properties of the methanol extract of Ruellia praetermissa Sceinf. ex. Lindau (Acanthaceae) in female rats. AB - In search for alternative drugs with pharmacological profile to replace hormone replacement therapy, the effects of MeOH extract of Ruellia praetermissa on the uterus and gestation in rats was investigated. 350 mg/kg/day of extract from days 1-9, 1-17 and 9-17 respectively, resulted in increase of the number of implantation sites (56 to 64) and the percentage of implantation (68.6 +/- 2.7 to 90.5 +/- 0.5%). There was also an increase in body weight (1-9 and 1-17) followed by a slight decrease (154 +/- 15.5 to 125 +/- 10) in the body weight at term. The number and the mean value of corpora lutea per female decreased from 25.4 +/- 1.6 to 14.00 +/- 1.6. The extract produced dose-related contraction of the isolated uterine muscle in vitro comparable to ACh. Atropine in doses from 3.4 x 10(-6) to 3 x 10(-3) microM antagonized the response of the uterus to ACh at 2 microM. It induced an increase (0.03 +/- 0.002 to 0.34 +/- 0.001 g) of the uterine weight comparable to that produced by using 3 microM estradiol (0.03 +/- 0.001 to 0.35 +/- 0.005 g). It could therefore be postulated that this extract possesses estrogenic and possible cholinergic effects. The estrogenic effect could have been generated by plant sterols (beta-sitosterol and stigmasterol) and flavonoids (luteolin and apigenin) while cholinergic effect could be due to iridoid glycosides. PMID- 11924765 TI - Complement modulating activity of Rwandan medicinal plants. AB - Forty-two ethanolic extracts of thirty-six Rwandan medicinal plants were investigated for their influence on complement-mediated hemolysis. The plants were selected on the base of their ethnomedicinal use in infections and autoimmune diseases. Eight plant extracts showed an inhibitory activity against the classical pathway of the complement system and ten plant extracts against the alternative pathway. Three plant extracts exhibited an interesting activity against both pathways, i.e. Aspilia pluriseta, Coleus kilimandschari, and Macaranga kilimandscharica (leaves and stem). Further study indicated that the complement inhibitory activity was not caused by chelation of bivalent cations or by direct action on the target erythrocytes. PMID- 11924766 TI - Antiviral activity of Rwandan medicinal plants against human immunodeficiency virus type-1 (HIV-1). AB - Selected plants used in Rwandan traditional medicine for the treatment of infections and/or rheumatoid diseases were investigated for antiviral activity in vitro against human immunodeficiency virus type-1 (HIV-1). Of the 38 tested 80% ethanolic extracts, belonging to plants of 21 different families only the extracts from the leaves of Aspilia pluriseta (Asteraceae) and Rumex bequaertii (Polygonaceae) had interesting selectivity indices (SI = ratio of the 50% cytotoxic concentration to the 50% effective antiviral concentration) higher than 1. Further fractionation of the initially antivirally inactive ethanolic extract of Tithonia diversifolia, however, led to an aqueous fraction with a high anti HIV-1 activity (SI > 461), indicating that the cytotoxicity of some plant components may mask the antiviral properties of the active plant substances in total plant extracts. PMID- 11924767 TI - Acute and chronic toxicity of Nigella sativa fixed oil. AB - We investigated the toxicity of the fixed oil of Nigella sativa L seeds in mice and rats through determination of LD50 values and examination of possible biochemical, hematological and histopathological changes. The acute toxicity of Nigella sativa fixed oil was investigated in mice. LD50 values, obtained by single doses, orally and intraperitoneally administered in mice, were 28.8 ml/kg body wt. p.o. [26.2-31.6] and 2.06 ml/kg body wt. i.p. [1.86-2.26], respectively. Chronic toxicity was studied in rats treated daily with an oral dose of 2 ml/kg body wt. for 12 weeks. Changes in key hepatic enzymes levels, including aspartate aminotransferase, alanine-aminotranferase, and gamma-glutamyltransferase and histopathological modifications (heart, liver, kidneys and pancreas) were not observed in rats treated with Nigella sativa after 12 weeks of treatment. The serum cholesterol, triglyceride and glucose levels and the count of leukocytes and platelets decreased significantly, compared to control values, while hematocrit and hemoglobin levels increased significantly. A slowing of body weight gain was also observed in Nigella sativa treated rats, as compared to control animals. The low toxicity of Nigella sativa fixed oil, evidenced by high LD50 values, key hepatic enzyme stability and organ integrity, suggests a wide margin of safety for therapeutic doses of Nigella sativa fixed oil, but the changes in hemoglobin metabolism and the fall in leukocyte and platelet count must be taken into consideration. PMID- 11924768 TI - Effect of Trichodesma indicum extract on cough reflex induced by sulphur dioxide in mice. AB - The effect of methanol extract of whole plants of Trichodesma indicum R. Br. has been investigated on sulphur dioxide (SO2) induced cough reflex in Swiss albino mice. The extract has demonstrated significant (p < 0.001) inhibition in frequency of cough in all the tested doses when compared with untreated control group. The effect persisted up to 90 min of its oral administration and also comparable to that of the effect exhibited by the standard drug (Codeine phosphate). This study confirmed the traditional use of this plant in the treatment of cough. Determination of underlying mechanism of beneficial effect is major topic requiring further comprehensive investigation. PMID- 11924769 TI - Safety and efficacy of ephedra and ephedrine for enhancement of athletic performance, thermogenesis and the treatment of obesity. PMID- 11924770 TI - Stevioside induces antihyperglycaemic, insulinotropic and glucagonostatic effects in vivo: studies in the diabetic Goto-Kakizaki (GK) rats. AB - Extracts of leaves from the plant Stevia rebaudiana Bertoni have been used in the traditional treatment of diabetes in Paraguay and Brazil. Recently, we demonstrated a direct insulinotropic effect in isolated mouse islets and the clonal beta cell line INS-1 of the glycoside stevioside that is present in large quantity in these leaves. Type 2 diabetes is a chronic metabolic disorder that results from defects in both insulin and glucagon secretion as well as insulin action. In the present study we wanted to unravel if stevioside in vivo exerts an antihyperglycaemic effect in a nonobese animal model of type 2 diabetes. An i.v. glucose tolerance test (IVGT) was carried out with and without stevioside in the type 2 diabetic Goto-Kakizaki (GK) rat, as well as in the normal Wistar rat. Stevioside (0.2 g/kg BW) and D-glucose (2.0 g/kg BW) were administered as i.v. bolus injections in anaesthetized rats. Stevioside significantly suppressed the glucose response to the IVGT in GK rats (incremental area under the curve (IAUC): 648 +/- 50 (stevioside) vs 958 +/- 85 mM x 120 min (control); P < 0.05) and concomitantly increased the insulin response (IAUC: 51116 +/- 10967 (stevioside) vs 21548 +/- 3101 microU x 120 min (control); P < 0.05). Interestingly, the glucagon level was suppressed by stevioside during the IVGT, (total area under the curve (TAUC): 5720 +/- 922 (stevioside) vs 8713 +/- 901 pg/ml x 120 min (control); P < 0.05). In the normal Wistar rat stevioside enhanced insulin levels above basal during the IVGT (IAUC: 79913 +/- 3107 (stevioside) vs 17347 +/- 2882 microU x 120 min (control); P < 0.001), however, without altering the blood glucose response (IAUC: 416 +/- 43 (stevioside) vs 417 +/- 47 mM x 120 min (control)) or the glucagon levels (TAUC: 5493 +/- 527 (stevioside) vs 5033 +/- 264 pg/ml x 120 min (control)). In conclusion, stevioside exerts antihyperglycaemic, insulinotropic, and glucagonostatic actions in the type 2 diabetic GK rat, and may have the potential of becoming a new antidiabetic drug for use in type 2 diabetes. PMID- 11924771 TI - The balancing act. PMID- 11924772 TI - Legal protection for rats, mice, and birds: long overdue and the right thing to do. PMID- 11924773 TI - The academic paper. PMID- 11924774 TI - A primer in epidemiologic methodology. AB - Epidemiology is defined as the study of the distribution and determinants of disease within populations. In addition to the requirements for disease surveillance, epidemiologic methods have numerous applications in laboratory animal science and can reveal important insights into the multifactoral mechanisms of disease, thereby aiding in the design of optimized intervention strategies. Observational approaches to data collection can be used to quantify the role of causal factors under natural circumstances, complementing the value of experimental studies in this field. The meaning and appropriate use of standard measures of disease frequency and exposure-disease relationships are reviewed, along with explanations of bias and confounding. Recommendations for reporting the methods and findings from this type of work in comparative medicine literature are presented. Aspects of model-based approaches to data analysis are introduced, offering further opportunities for gaining needed information from epidemiologic study of problems in laboratory animal medicine and management. PMID- 11924775 TI - Relations among measures of body composition, age, and sex in the common marmoset monkey (Callithrix jacchus). AB - Few studies of body composition have been done in New World primates. In the study reported here, four methods of assessing body composition (body weight, anthropometry, labeled-water dilution, and total body electroconductivity) were compared in 20 marmosets, aged 0.96 to 7.97 years. Males and females did not differ in any measure (P > 0.05). Body weight ranged from 272 to 466 g, and body fat estimates varied from 1.6 to 19.5%. Strong positive correlations were observed between total body water and total body electroconductivity (R2 = 0.77), body weight and fat-free mass (males R2 = 0.95; females R2 = 0.91), and body weight and fat mass (males R2 = 0.86; females R2 = 0.85; P < 0.01). Male and female slopes were equivalent (P > 0.05) for the regressions of fat and fat-free mass against body weight. Positive correlations also were observed between girth measures and fat-free mass (R2 = 0.48 to 0.78) and fat mass (R2 = 0.60 to 0.74; P < 0.01). A good second- order polynomial relationship was observed between age and fat-free mass for the combined sample (R2 = 0.64). Results indicated that: subjects were lean; there was no sexual dimorphism relative to measures; body weight provided a reliable estimate of fat and fat-free mass; and within-subject body weight changes reflected a similar relationship between body weight and fat free mass as did that across subjects. PMID- 11924776 TI - Effects of buprenorphine on immunogenicity and protective efficacy in the guinea pig keratoconjunctivitis model (Sereny test). AB - Shigellosis is a disease of global proportions, with an estimated 164.7 million episodes annually throughout the world as well as an estimated 1.1 million associated mortalities in developing countries. Due to increasing incidence, and continued emergence of multi-drug resistant strains, Shigella vaccine development is considered a top public health priority. The guinea pig keratoconjunctivitis model, the basis for the Sereny test, remains the most reliable in vivo indicator of virulence of Shigella strains and immunogenicity and protective efficacy of Shigella vaccine candidates. The model is effective in evaluating the ability of Shigella strains to invade the corneal epithelia of guinea pigs and spread to contiguous cells, with the more virulent strains causing ulcerative keratoconjunctivitis. However, analgesia is not routinely used to relieve this painful condition because of potential immunomodulation and confounding of experimental results. The objective of the study reported here was to evaluate use of buprenorphine hydrochloride as an analgesic during the Sereny test. Local and systemic immune responses were measured in guinea pigs given buprenorphine versus those responses in controls. Results of this study suggest that buprenorphine, administered at an analgesic dose of 0.05 mg/kg of body weight twice daily, can be successfully used with the model without significantly affecting immunologic evaluation of Shigella vaccine candidates. However, in buprenorphine-treated animals, there was a significant increase in the amount of mucopurulent ocular discharge, requiring frequent cleaning of the affected eyes. Additionally, animals treated with buprenorphine had significant reduction in body weight, in comparison with saline controls. PMID- 11924778 TI - Monoclonal antibodies reactive with dendritic cells of Mongolian gerbils. AB - Mongolian gerbils (Meriones unguiculatus) serve as an valuable model animal for several infectious diseases of medical and veterinary importance. Reagents available for characterization of the immune response of Mongolian gerbils are strictly limited. We describe three novel murine monoclonal antibodies (mAbs) to dendritic cells of Mongolian gerbils. These include HUSM-M.g.11 of IgG2b isotype, HUSM-M.g. 20 of IgG2a isotype, and HUSM-M.g.30 of IgG1 isotype. All of these mAbs had an identical profile of immunohistochemical reactions with various tissues taken from immune-naive Mongolian gerbils, and were intensively expressed on dendritic cells, including epidermal Langerhans cells, B-cell follicles, and the thymic reticulum. Positive reactions of the epidermis and intestinal mucosa with these mAbs were induced by cutaneous or intestinal infections with parasites. Competitive enzyme-linked immunosorbent assay and immunoblot analysis (western blotting) indicated that all of these mAbs recognize an identical peptide epitope on a molecule with approximate molecular mass of 29 kDa. These data suggest that the mAbs recognize major histocompatibility complex class-II molecules of gerbils. Use of described mAbs would facilitate characterization of immune responses as well as investigations on host responses to infections of medical and veterinary importance, using the gerbil model. PMID- 11924777 TI - Increased dietary fat prevents sleep deprivation-induced immune suppression in rats. AB - BACKGROUND AND PURPOSE: Fatty acid composition of rodent diets can affect baseline immune function as measured in vitro and in vivo. Stress, in a variety of forms, can also affect immune function. Possible interaction between diet and other stressors has not been fully explored. We examined the interaction between sleep deprivation stress and dietary fatty acid composition in altering lymphocyte responses to mitogen stimulation. METHODS: Rats were fed diets containing various sources of fatty acids, then were subjected to sleep deprivation. Splenocytes were harvested and assayed for responsiveness to various mitogens, using a 72-h proliferation assay. RESULTS: Rats subjected to sleep deprivation experienced significant suppression of in vitro proliferative response to various mitogens. This immune suppression was dependent on duration of sleep deprivation. Feeding sleep deprived rats a diet enriched in fatty acids abrogated the effect of sleep deprivation. CONCLUSIONS: The fat content of rodent diets can have a marked effect on baseline and stress-modulated immune responses. PMID- 11924779 TI - Evaluation of fentanyl transdermal patches in rabbits: blood concentrations and physiologic response. AB - In the study reported here, we sought to evaluate transdermal fentanyl patches for their ability to achieve detectable plasma concentrations with minimal adverse effects in New Zealand White rabbits. Fentanyl patches were applied to the dorsum after removing hair either by clipping or by application of a depilatory agent. Blood samples were collected every 12 h for a total of 96 h (24 h after patch removal) for determination of plasma fentanyl concentration. At those times, rabbits were assessed for changes in body temperature, heart rate, respiratory rate, and body weight. In rabbits with clipped hair, where rapid hair re-growth was not a mitigating factor, mean plasma fentanyl concentration reached a mean (+/- SEM) peak of 1.11 +/- 0.32 ng/ml at 24 h, decreased to 0.77 +/- 0.21 ng/ml at 72 h, and was negligible at 96 h. In rabbits with depilated hair, peak concentration was obtained at 12 h (6.7 +/- 0.57 ng/ml) and decreased gradually to 0.27 +/- 0.06 ng/ml at 72 h. In a second group of fentanyl-treated rabbits in which hair started growing back within 24 h, plasma fentanyl concentration was not detectable. Control and fentanyl-treated rabbits with clipped hair had no effect from the experimental manipulations other than slight loss in body weight. In the depilatory group, two rabbits appeared moderately sedated during the initial 12-h period, and had decreased respiratory rate for 24 h. In conclusion, rabbits tolerate the transdermal fentanyl patch well. Hair regrowth in rabbits may present a complicating factor that impedes dermal absorption of fentanyl. The application of a depilatory agent lead to early and rapid absorption of fentanyl causing undue sedation in some rabbits and lack of sustained plasma concentrations for the desired three-day period. PMID- 11924780 TI - New rat model for attention deficit hyperactive disorder (ADHD). AB - PURPOSE: In a strain of the Long-Evans Cinnamon (LEC) rats, we found spontaneously hyperactive animals designated as "wiggling," and established a congenic wiggling (Wig) rat by transferring the gene from the LEC to the Wistar King-Aptekman/Hokkaido (WKAH) strain. We evaluated the feasibility of the Wig rat for an animal model of human attention deficit hyperactive disorder (ADHD). METHODS: Mode of inheritance was examined by use of linkage analyses. Motor activity, behavior, and working memory were assessed by use of electric digital counters, open field test, and Y-maze and water-maze tests. RESULTS: The abnormal behavior, including hyperactivity, was transmitted in autosomal recessive mode. Diurnal and nocturnal motor activity of 12- to 14-week-old Wig rats was markedly higher than that of controls, and this hyperactivity was more prominent during nighttime than daytime. Ambulation in the open-field test was significantly increased in Wig rats, but rearing was decreased in Wig rats, compared with controls. Results of the Y-maze tests indicated that spontaneous alternation behavior was significantly impaired in Wig rats, although there was no difference in the total arm entries. The water-maze test could not be performed because, when exposed to water, Wig rats panicked and almost drowned. CONCLUSIONS: Wig rats are hyperactive and have impaired working memory and impulsive behavior, as assessed by the motor activity and open-field tests and the Y-maze test, and these abnormalities are transmitted by a single gene with Mendelian pattern. Wig rats represent an excellent animal model of human ADHD. PMID- 11924781 TI - Permanent jejunal fistula: promising method for obtaining small intestinal chyme without disturbing intestinal function. AB - Accurate information on changes in small intestinal microflora in dogs is rather limited because of difficulties in obtaining samples of small intestinal chyme. In the study reported here, intussuscepted nipple valves were surgically placed into the jejunum of seven laboratory beagles to obtain intestinal juice samples. The influence of the fistula on intestinal motility was determined by use of barium-impregnated polyethylene spheres (BIPS) and on microflora by use of bacterial culturing. The BIPS were fed two weeks before surgery and again five weeks after surgery. Bacterial samples were collected before (fecal samples), during (small intestinal samples) and 11 weeks after surgery. There were no surgical complications, and the animals tolerated the fistula well. Mean orocolic transit percentage was 93% before and 83% after surgery, and notable changes in gastrointestinal motility were not seen, except in one dog. The surgery did not markedly alter the bacterial flora in feces. Microflora did change in small intestinal samples; however, methodologic factors may explain most of these differences. In conclusion, the nipple valve is a promising method that creates easy and safe long-term access to the jejunum and appears not to have an influence on intestinal function. PMID- 11924782 TI - Campylobacter-induced enteritis and diarrhea in captive cotton-top tamarins (Saguinus oedipus) during the first year of life. AB - A prospective study of 43 cotton-top tamarins, from infancy to 6 to 17 months of age, was conducted to determine the epidemiology of Campylobacter spp. infection. Nine infants followed for one year in an isolation unit, where attendants wore protective clothing, did not become infected. In the main facility where 32 of 34 animals had repeated infections with C. coli, 6% of the infections developed initially in incubators, 66% in the nursery room, and 28% after transfer to the main colony. Fifteen of these tamarins also were infected with C. jejuni. Twenty percent of the infections developed initially in the nursery room and 80% in the colony. Polyacrylamide gel electrophoresis analysis of C. jejuni cultures revealed multiple reinfections with different strains. Both types of infections were most prevalent between 3 and 9 months of age. Campylobacterjejuni infection developed most frequently between April and June and C. coli infection developed between October and December. In the nursery, diarrhea developed most frequently at times when there was no infection with Campylobacter spp. Forty percent of animals with diarrhea in the nursery had C. coli and none had C. jejuni, whereas, in the colony, 49% had C. jejuni and 11% had C. coli infections. There was no association between these infections and diet or idiopathic colitis. PMID- 11924783 TI - Gastric dilatation syndrome associated with chronic nephropathy, hypergastrinemia, and gastritis in mice exposed to high levels of environmental antigens. AB - Gastric dilatation (GD) has been observed in Tac:(SW)fBR surveillance mice, with mean age of 10 months, that are exposed to high levels of environmental antigens during routine exposure to dirty bedding. The aim of the study reported here was to determine whether GD was associated with other systemic conditions affecting mice. Three groups of nine animals including-surveillance mice not exposed to dirty bedding (control), surveillance mice with out GD (NGD), and surveillance mice with GD (group GD)-had mean stomach weight with ingesta of 0.5 +/- 0.02 g, 1.09 +/- 0.07 g (P < 0.0001), and 2.54 +/- 0.4 g (P < 0.0001), respectively. Mean serum creatinine concentration was significantly higher in GD (1.6 +/- 0.25 mg/dl), compared with NGD (0.17 +/- 0.22 mg/dl, P < 0.0001) and control (0.2 +/- 0.16 mg/ dl, P < 0.0001) mice. In addition, lesions consistent with severe chronic nephropathy and mild gastritis were common in GD, compared with NGD and control mice. Finally, serum amidated gastrin concentration was significantly high in GD (179.37 +/- 53.86 pM, P < 0.03) and NGD (264.89 +/- 115.89 pM, P < 0.009), compared with control (60.77 +/- 8.39 pM) mice. Gastric dilatation syndrome is associated with chronic nephropathy, hypergastrinemia, and gastritis in surveillance mice exposed to high levels of environmental antigens. PMID- 11924784 TI - Laboratory animal medicine in a time of crisis. PMID- 11924785 TI - The balancing act/legal protection for rats, mice and birds. PMID- 11924786 TI - Sheep model in orthopedic research: a literature review. AB - The aim of the study reported here was to provide some basic and general information on the suitability of an experimental sheep model for conducting in vivo orthopedic studies. The authors have classified the fundamental aspects that should be carefully evaluated when using sheep as an experimental model in orthopedic research: factors strictly related to bone anatomy and formation; and factors strictly affecting bone physiology, such as gastrointestinal mineral and vitamin absorption, and reproductive cycle. Future investigations should address all of the aspects highlighted, since there is no animal with the same anatomic, biochemical, physiologic, and biological characteristics as those of human beings. Moreover, useful data for treating orthopedic patients are based not only on good planning and study design, but also on perfect knowledge of the animal used and the differences between the model and the human being. The authors hope that this report will contribute to extrapolation of reliable data for use of sheep in the orthopedics field. PMID- 11924787 TI - Primer for non-immunologists on immune-deficient mice and their applications in research. AB - Studies of immune deficiencies have a history as long as that of immunology. However, reports of two key spontaneous recessive mutations in mice (nude in 1966 1968 and scid in 1983) laid the foundations for widespread application of immune deficient rodents to a broad range of research topics. More recently, technologies modifying the mouse genome by transgenesis, gene ablation and crossbreeding for lines with multiple immune deficits have provided a large number of new types of immunologically impaired mice. The primary goals of this overview are to help non-immunologists understand key differences between some of the immunodeficient strains, develop an appreciation for the value of information derived from immunodeficient mouse-based research and to encourage expanded, creative use of these specialized research animals. Secondary goals are to promote greater awareness of unexpected outcomes that can arise when working with genetically immune-deficient mice, the need for vigilance in maintaining these research animals, and the care required in interpretation of the data that immune deficient modeling provides. Two illustrations on developing appropriate immune deficient animal models for a new research application conclude the review. PMID- 11924788 TI - Establishment of a deer mouse (Peromyscus maniculatus rufinus) breeding colony from wild-caught founders: comparison of reproductive performance of wild-caught and laboratory-reared pairs. AB - The deer mouse (Peromyscus maniculatus) is a natural reservoir for several human pathogens, but little is known about the mechanisms by which such pathogens are maintained in nature. As a first step toward developing a colony of deer mice that were permissive for infection with Sin Nombre (SN) hantavirus, we collected 68 wild P. maniculatus rufinus from central New Mexico. Mice from this cohort were used to establish 26 breeding pairs, of which 85% were fertile. In subsequent generations, fertility decreased slightly to 73% (N = 59) in laboratory-reared F1 and F2 pairs. Wild-caught females delivered 7.2 litters on average (range, 1 to 18), whereas laboratory-reared pairs delivered 5.5 (range, 1 to 13). The average time between pairing and first litter was 106 days for wild caught animals, whereas that for laboratory-reared pairs was 71 days. None of the pairs displayed a seasonal breeding preference. Cannibalistic behavior increased from 5% in founders to 26% in laboratory-reared pairs. Mean litter size for wild caught females was 4.3, whereas that for laboratory-reared dams was 4. Founding animals have been maintained in captivity for longer than 2 years, with only 2 deaths (4.8%). Our colony is competent for infection with SN virus. Thus, it should be useful for testing of models for maintenance of SN virus in wild rodents, and other aspects of the virus-host relationship. PMID- 11924789 TI - Differentiation of mouse hepatitis viruses in animal facilities in Japan by use of nucleotide analysis of the nucleocapsid gene. AB - The nucleotide sequences of the coding region of the nucleocapsid (N) gene of 12 mouse hepatitis virus (MHV) strains recently found in animal facilities in Japan were analyzed. Nucleotide sequencing was performed directly on polymerase chain reaction (PCR) products amplified by reverse transcription (RT) and polymerase chain reaction (RT-PCR) analysis from fecal samples or isolated viruses. Phylogenetic analysis of these MHV strains along with those reported previously indicated that sequence analysis of the N gene was a useful tool for differentiation of MHV strains,although most MHV strains in Japanese facilities were phylogenetically close. Results suggested that interchange of mice infected with MHV among facilities provided opportunities of introduction of MHV into otherwise MHV-free facilities and that the source of MHV infection could be traced by use of nucleotide analysis of the N gene. PMID- 11924790 TI - Detection of rodent parvoviruses by use of fluorogenic nuclease polymerase chain reaction assays. AB - Polymerase chain reaction (PCR) assays have proven useful for detection of rodent parvoviruses in animals and contaminated biological materials. Fluorogenic nuclease PCR assays combine PCR with an internal fluorogenic hybridization probe, eliminating post-PCR processing and potentially enhancing specificity. Consequently, three fluorogenic nuclease PCR assays were developed, one that detects all rodent parvoviruses, one that specifically detects minute virus of mice (MVM), and one that specifically detects mouse parvovirus 1 (MPV) and hamster parvovirus (HaPV). When rodent parvoviruses and other rodent DNA viruses were evaluated, the rodent parvovirus assay detected only rodent parvovirus isolates, whereas the MVM and MPV/HaPV assays detected only the MVM or MPV/ HaPV isolates, respectively. Each assay detected the equivalent of 10 or fewer copies of target template, and all fluorogenic nuclease PCR assays exceeded the sensitivities associated with previously reported PCR assays and mouse antibody production testing. In addition, each fluorogenic nuclease PCR assay detected the targeted parvovirus DNA in tissues obtained from mice experimentally infected with MVM or MPV. Results of these studies indicate that fluorogenic nuclease PCR assays provide a potentially high-throughput, PCR-based method to detect rodent parvoviruses in infected mice and contaminated biological materials. PMID- 11924791 TI - Variation of serum alpha2-macroglobulin concentration in healthy rats and rats inoculated with Staphylococcus aureus or subjected to surgery. AB - BACKGROUND AND PURPOSE: The aim of the study reported here was to investigate variations in the serum concentration of alpha2-macroglobulin (alpha2M) from healthy rats and rats inoculated with Staphylococcus aureus or subjected to surgery. METHODS: Concentration of alpha2M was measured by use of an enzyme linked immunosorbent assay. RESULTS: Serum alpha2M in healthy rats at intervals of 3 h, 1 day, and 1 week ranged from 17.5 to 38.0 (mean +/- SD, 27.2 +/- 6.6) microg/ml, 15.8 to 48.2 (27.9 +/- 8.7) microg/ml, and 17.2 to 39.6 (23.9 +/- 5.9) microg/ml, respectively. Concentrations of alpha2M did not undergo significant variations within individuals or between rats. Serum alpha2M concentration increased at one day and peaked two days after inoculation with S. aureus or being subjected to surgery. Peak concentration was eight to 33 times preinoculation values after inoculation with S. aureus, four to 25 times pre surgical values when rats were subjected to castration, and seven to 28 times pre surgical values when rats were subjected to oophorohysterectomy, respectively. CONCLUSIONS: Physiologic variation in the concentration of alpha2M in rats was not found. Induction of alpha2M in rats inoculated with S. aureus or subjected to surgery was documented. PMID- 11924792 TI - Similarity of bone ingrowth in rats and goats: a bone chamber study. AB - Bone ingrowth has been studied extensively in rats by use of bone chambers. However, it is not known whether results in small animals, with respect to bone ingrowth processes, are similar in large animals, in which more realistic models are often used. Since the metabolic rate in small animals is, in general, higher than that in larger species, we hypothesized that bone ingrowth in chambers develops more rapidly in small animals. Therefore, identical bone chambers were placed in the tibias of rats and goats. After 6 and 12 weeks, histologic and histomorphometric examinations were carried out to measure bone and tissue ingrowth distances. Bone ingrowth was higher in both species at 12, compared with 6 weeks (P < 0.01). Tissue ingrowth in general (including soft tissue) was less in rats than in goats at both time periods (P < 0.001). However, bone ingrowth did not differ between species. Thus, when differences in size of an osseous defect are corrected for, there seems to be only little influence of differences in body size. PMID- 11924793 TI - The pig as a model for excisional skin wound healing: characterization of the molecular and cellular biology, and bacteriology of the healing process. AB - A pig model of wound healing was developed by excision of 2-cm-diameter full thickness skin in young Yorkshire pigs. The results indicated that wound re epithelialization in this animal model took an average of 20 days. Analysis of cellular change was assessed by use of DNA quantification and determination of apoptotic cells in tissue sections. The results indicate that RNA and DNA contents paralleled each other throughout the healing process, and observed changes in the pattern of RNA and DNA content of the scar tissues were consistent with cell loss due to apoptosis in this model. Expression of mRNA for relevant genes was assessed by use of semiquantitative reverse transcription-polymerase chain reaction (RT-PCR) analysis, using porcine specific primer sets and RNA isolated from normal skin and specimens obtained at various times after wounding. The mRNA values for tumor necrosis factor-alpha (TNF-alpha), connective tissue growth factor (CTGF), insulin-like growth factor II (IGF-II), and decorin were significantly high at specific times after wounding, but mRNA values for the transcription factors (c-fos and c-jun) were significantly decreased. Quantitative bacteriologic results indicated that the total bacterial count in this animal model reached 10(9) colony-forming units (CFU)/g, with the highest value at post-wounding day 7, and Pseudomonas aeruginosa and Staphylocococci aureus were the most common bacteria detected in this model. Further definition of this model should identify unique points in the healing process, and such information could lead to development of therapeutic interventions to improve skin wound healing. PMID- 11924794 TI - Development of a novel intestinal and vascular access port (IVAP) rabbit model to study regiospecific oral absorption pharmacokinetics. AB - BACKGROUND AND PURPOSE: The limited availability and cost of many drugs prohibits routine use of the previously developed intestinal and vascular access port (IVAP) canine model by our group. A lower animal species model such as the rabbit is suitable for implanting intestinal and vascular access ports for investigating regiospecific intestinal absorption and hepatic elimination while requiring significantly lower doses of drugs. In addition, expression of certain cytochrome P450 enzymes and apical secretory and absorptive transporters in rabbit intestine is similar to that in humans making the rabbit a suitable model. METHODS: Individual 5-F Silastic catheters were placed in the proximal or distal portion of the small intestine or the colon of subject animals, while a 5-F Heparin Coated Polyurethane (HCP) catheter was implanted in the portal vein of each subject. The catheters were tunneled out of the abdomen and attached to separate subcutaneous access ports along the spine. The animals were allowed a two-week minimum recovery period prior to being used in pharmacokinetic studies. RESULTS AND DISCUSSION: After some initial difficulties, rabbits with IVAP implants proved to be an efficient and dependable model for investigating intestinal and hepatic extraction of drugs. Fluoroscopic visualization of intestinal and portal venous catheters indicated that surgically implanted catheters did not interfere with gastrointestinal motility or blood flow into the liver, respectively. Acute pH studies in the proximal portion of the small intestine were consistent with normal GI motility patterns. PMID- 11924795 TI - Coxiella burnetii infection in C.B-17 scid-bg mice xenotransplanted with fetal bovine tissue. AB - Two from a group of approximately 50 C.B-17 scid-bg mice were examined because of lethargy, dehydration, and rough coat. Three months prior to development of clinical signs of disease, mice of this study had been surgically implanted with fetal bovine liver, thymus, and lymph node. At necropsy, marked splenomegaly and mild hepatomegaly were observed in both animals. Large areas of necrosis and inflammation, with associated intracytoplasmic granular basophilic inclusions, were observed in histologic sections of multiple organs. Aerobic and anaerobic culturing of the liver yielded negative results. Six months after the initial case, four more reconstituted scid-bg mice from a different fetal donor had identical clinical, gross, and histologic signs of disease. To determine whether the basophilic inclusions represented an infective agent, 4-month-old immune naive C.B-17 scid-bg mice were inoculated intraperitoneally with a liver and spleen homogenate from an affected mouse. Two weeks after inoculation, mice developed clinical signs of disease and lesions identical to those seen in the signal mice. On ultrastructural examination of the liver, pleomorphic bacteria were found in large cytoplasmic vacuoles of hepatocytes. Bacterial DNA was amplified from the liver, using primers that amplify a segment of the 16S rRNA gene from many bacterial species. Sequencing of the polymerase chain reaction (PCR) product revealed gene sequence identical to that of Coxiella burnetii, the agent of Q-fever. These results highlight the need to consider infective agents of the donor species when working with xenografted animals. PMID- 11924796 TI - High mortality in a large-scale zebrafish colony (Brachydanio rerio Hamilton & Buchanan, 1822) associated with Lecythophora mutabilis (van Beyma) W. Gams & McGinnis. AB - Zebrafish (Brachydanio rerio) have become an important model system for studying vertebrate embryonic development and gene function through manipulation of genotype and characterization of resultant phenotypes. An established research zebrafish colony without substantial disease problems for more than 7 years of operation began experiencing appreciable mortalities in November of 1997. Young fish (fry), from five to 24 days after hatching, spontaneously developed elongate strands of organic material protruding from the mouth, operculum, and anal pore, leading workers in the laboratory to describe the infected fish as "bearded." Unlike typical freshwater fish fungal infections, the skin surface did not have evidence of fungal colonization. The disease was associated with progressive lethargy, reduced feeding, and subsequent mortality. From 10 to 100% of the fry in a given tank were affected. Initial examination indicated that the biofilm around the head of affected fry consisted of bundles of septate fungal hyphae, large numbers of mixed bacterial populations, and protozoans. Environmental samples of air and water in the laboratory were obtained to ascertain the source of the infective agent and to isolate and identify the fungus. A fungus identified as Lecythophora mutabilis was isolated repeatedly from infected fish and water samples from infected fish tanks, and from the main laboratory water supply tanks, but not from laboratory air. Some biofilm beards on fish were found to consist of relatively pure bacterial populations, and beards on occasional fish examined in the later part of the study consisted of hyphae and spores of the oomycete genus Aphanomyces. Lecythophora mutabilis did not invade tissues; however, elimination of the epizootic correlated with reduction in the number of L. mutabilis conidia in the water following modification of the laboratory water system by use of new filtration and sterilization systems. We conclude that the dense hyphal strands of L. mutabilis composing the predominant biofilm type, along with mixed bacteria and protozoa, contributed to the die-off in young fry by occluding the oral cavity and/or gills, leading to starvation and/or asphyxiation. PMID- 11924798 TI - Use of genetically modified mouse models for evaluation of carcinogenic risk: considerations for the laboratory animal scientist. AB - There has been increasing interest in the use of selected genetically modified (GM) mouse models for the testing of chemicals to determine their carcinogenic potential. GM mouse models are believed to be useful tools that offer mechanistically relevant insights for understanding and predicting the human response to chemical exposure. They have been proposed as alternatives to the traditional 2-year mouse oncogenicity bioassay. In this overview we will review the GM mouse models that have been proposed as bioassay alternatives and present some of the key laboratory animal science challenges that need to be considered when using these unique animals. PMID- 11924797 TI - Persistent transmission of mouse hepatitis virus by transgenic mice. AB - Variation in susceptibility to viral infection is well documented across mouse strains. Specific combinations of viral strains and murine hosts may favor viral infection and disease, and could potentially allow the unexpected development of chronic, persistent, or latent infections. In some genetically modified strains of mice, immune function and perhaps other physiologic or metabolic systems may be substantially or marginally impaired. In the case study reported here, we document the apparent persistent transmission of mouse hepatitis virus (MHV) over a two-year period by MHV-seropositive transgenic mice. Transmission occurred via direct contact with seropositive mice and exposure to contaminated bedding. However, MHV was not detected at diagnostic laboratories by use of viral isolation or reverse transcriptase-polymerase chain reaction (RT-PCR) analysis of tissues from MHV-seropositive animals. Our observation, together with the constantly expanding varieties of immune-impaired or poorly characterized murine hosts and the burgeoning dissemination of these animals throughout the biomedical research community, indicate that unexpected pathophysiologic presentations of common murine viral diseases may present new challenges to the biomedical research community in the future. PMID- 11924799 TI - Evaluation of diagnostic methods for Helicobacter bilis infection in laboratory mice. AB - Disease-susceptible (C3H) and -resistant (B6) immunocompetent and immunodeficient (C3H-scid and B6-rag1) mice were examined up to 10 weeks after inoculation with Helicobacter bilis (a prototype species of proven virulence). Infection was monitored weekly by use of fecal culture, polymerase chain reaction (PCR) nucleic acid amplification, membrane extract enzyme-linked immunosorbent assay (ELISA), and histologic examination. All mice became infected by three to five weeks after inoculation, on the basis of results of culture and PCR analysis of feces. The PCR analysis was more sensitive than culture at determining infection status, particularly during early infection. None of the mice had evidence of disease by week 10. Immunoglobulin G seroconversion was detectable in C3H mice by week eight and in B6 mice by week nine. Results indicated that culture and PCR analysis are more sensitive than is membrane extract ELISA serologic testing for detecting early infection in individual mice, regardless of genotype or immune status. Results underscore the need for improved seroassays for this important group of murine pathogens. PMID- 11924800 TI - Long-term colonization levels of Helicobacter hepaticus in the cecum of hepatitis prone A/JCr mice are significantly lower than those in hepatitis-resistant C57BL/6 mice. AB - Helicobacter hepaticus infection causes hepatitis in A/JCr mice but mild or no disease in C57BL/6 mice. Colonization of H. hepaticus in the cecum of experimentally infected A/JCr and C57BL/6 mice was quantified by use of real-time polymerase chain reaction (PCR) analysis with primers for the H. hepaticus cdtB gene and mouse 18srRNA. Eight-week-old mice were experimentally (n = 48) or sham (n = 24) infected with H. hepaticus, then were necropsied 6 months after infection. Liver specimens from experimentally infected mice had negative results of PCR analysis for H. hepaticus; thus, real-time quantification was not attempted. Quantitative PCR analysis of H. hepaticus in cecal specimens indicated that C57BL/6 mice were colonized to a greater extent than were A/JCr mice (P < 0.006). Appreciable typhlitis was not observed, but was consistent with that of previous reports; A/JCr mice developed more severe parenchymal necrosis, portal inflammation, and phlebitis in the liver (P < 0.0001), with mild disease observed in infected C57BL/6 mice. Thus, hepatitis in A/JCr mice caused by H. hepaticus infection is associated with significantly lower colonization levels of H. hepaticus in the cecum, compared with those of hepatitis-resistant C57BL/6 mice. Host responses of A/JCr mice that limit cecal colonization with H. hepaticus may have important roles in the pathogenesis of hepatic lesions. PMID- 11924801 TI - High intake of selenium, beta-carotene, and vitamins A, C, and E reduces growth of Helicobacter pylori in the guinea pig. AB - PURPOSE: Helicobacter pylori is a human gastroduodenal pathogen associated with type-B gastritis and gastric cancer. Low gastric tissue antioxidant levels are believed to increase the risk of developing gastric cancer. We investigated whether dietary antioxidant levels protect against infection and type-B gastritis in H. pylori-infected guinea pigs. METHODS: Dunkin-Hartley guinea pigs infected for 6 weeks with H. pylori were fed diets with various antioxidant levels. Stomach specimens were cultured, and gastritis was graded from 0 to 3. RESULTS: Supplementation with vitamins A, C, and E and with selenium yielded H. pylori recovery from 17% of challenged animals, compared with 43% of those fed a control diet. Gastritis was scored at 0.33 and 0.93, respectively. Supplementation with only vitamin C or astaxanthin had less effect on gastritis and recovery rate. In a second experiment, gastritis score in a group given vitamins A, C, E, and selenium and beta-carotene was 2.25 and in a control group, it was 2.57. The H. pylori recovery rate was 75 and 100%, respectively, with fewer colonies from animals given antioxidant supplementation (P < 0.05). CONCLUSIONS: A combination of antioxidants can protect against H. pylori infection in guinea pigs. In animal studies, antioxidant intake should be low to optimize development of H. pylori associated disease. Furthermore we established that H. pylori causes severe gastritis in guinea pigs. PMID- 11924802 TI - Rhesus macaque model for ovarian cancer chemoprevention. AB - PURPOSE: The objective of the study reported here was to explore whether a nonhuman primate model could be developed for chemoprevention of ovarian cancer. METHODS: An initial feasibility trial was done with three monkeys to determine tolerance for these drugs and for acquisition of surgical ovarian biopsy specimens. In the study, 19 female adult Macacca mulatta (rhesus macaques) were given fenretinide (4HPR) oral contraceptive (OCP), the combination of 4HPR+OCP, or no medication for three months. Laparotomy was performed before and after drug administration, and ovarian biopsy specimens were obtained to evaluate the potential for this animal as a model for ovarian cancer chemoprevention, as well as evaluating fluorescence spectroscopy and other potential biomarkers for ovarian cancer prevention studies. RESULTS: The monkeys tolerated the drugs, surgeries, and acquisition of multiple ovarian biopsy specimens with resultant minimal morbidity. On initial data analysis, fluorescence spectroscopy was the marker that appeared the most promising. CONCLUSIONS: On the basis of results of this study, this model merits further investigation. The rhesus monkey is an excellent candidate for a nonhuman primate model for ovarian cancer chemoprevention. PMID- 11924803 TI - Cyclosporine A-induced mammary hyperplasia and hyperprolactinemia in New Zealand White rabbits. AB - PURPOSE: To investigate the potential activity of cyclosporin A (CsA) to induce mammary hyperplasia in New Zealand White (NZW) rabbits. METHODS: Female NZW rabbits were used throughout experiments. To simulate the conditions of immunosuppression, CsA (10 mg/kg of body weight/d) was administered intravenously on a daily basis for 14 days and methylprednisolone (5 mg/kg/d) was administered on the first two days. The CsA (10 mg/kg/d) also was administered without methylprednisolone for 14 days to another cohort of rabbits. Mammary tissue of each rabbit was palpated and serially measured during this treatment period. The CsA was discontinued, and rabbits were monitored for 14 more days during the washout period. Sequential plasma concentrations of prolactin, 17beta-estradiol, and progesterone in each blood sample were determined by use of radioimmunoassay. RESULTS: All NZW rabbits treated with CsA and methylprednisolone for immunosuppression consistently developed striking mammary tissue hyperplasia. At the end of treatment with CsA and methylprednisolone, mammary glands had extensive changes consistent with actively lactating glands. Similar but less extensive hyperplasia developed in response to CsA alone. Plasma concentration of prolactin increased during treatment and decreased during the washout period. Plasma concentration of 17beta-estradiol increased during treatment and continued to increase during the washout period. Plasma progesterone concentration decreased at the end of treatment. On discontinuation of CsA, mammary hyperplasia regressed. CONCLUSIONS: Cyclosporine A, with or without methylprednisolone, induces mammary hyperplasia and hyperprolactinemia in NZW rabbits. This rabbit model may be a reliable in vivo system by which to study immunosuppressant induced structural and functional changes of mammary glands similar to those observed in humans. PMID- 11924804 TI - Parameters of glucose and lipid metabolism in the male Gottingen minipig: influence of age, body weight, and breeding family. AB - The pig is useful as a model for human physiology and pathophysiology and could be an important supplement to the many available rodent models of diabetes mellitus. Due to their small size, Gottingen minipigs are especially suitable for long-term studies. The aim of the study reported here was to establish reference values for a range of glucose and lipid homeostasis parameters of interest that could be used to identify possible diabetes-prone male Gottingen minipig individuals, families, or age groups. Plasma samples from nonfed animals were analyzed for glucose, leptin, fructosamine, insulin, C-peptide, triglyceride, free fatty acids, and total cholesterol values. Breeding family had significant effects only on plasma triglyceride concentrations (P < 0.001). Plasma concentrations of glucose (P = 0.012), fructosamine (P < 0.001) and triglycerides (P < 0.001) increased significantly with age, whereas total cholesterol concentration decreased significantly (P = 0.001) with age. Age did not influence other parameters. In conclusion, glycemia and insulinemia increased with age and body weight, possibly indicating a small deterioration in insulin sensitivity with age. It is, therefore, hypothesized that older, compared to younger animals may be more useful in the development of a model of type-2 diabetes mellitus. Furthermore, on the basis of decrease in cholesterol concentration with age, animals fed ad libitum with possibly a high calorie diet might be even more useful in the development of a type-2 diabetes mellitus model. PMID- 11924805 TI - Optimization of intraperitoneal injection anesthesia in mice: drugs, dosages, adverse effects, and anesthesia depth. AB - PURPOSE: The goals of the study were to find a safe intraperitoneal injection anesthesia protocol for medium-duration surgery in mice (e.g., embryo transfer/vasectomy) coupled with a simple method to assess anesthesia depth under routine laboratory conditions. METHODS: Eight anesthetic protocols consisting of combinations of dissociative anesthetics (ketamine, tiletamine), alpha2-agonists (xylazine, medetomidine), and/or sedatives (acepromazine, azaperone, zolazepam) were compared for their safety and efficacy (death rate, surgical tolerance), using observations and reflex tests. The four best protocols were further evaluated during vasectomy: physiologic measurements (respiratory rate, electrocardiogram, arterial blood pressure, body temperature, blood gas tensions, and acid-base balance) were used to characterize the quality of anesthesia. The reactions of physiologic parameters to surgical stimuli were used to determine anesthesia depth, and were correlated with reflex test results. RESULTS: The protocol with the highest safety margin and the longest time of surgical tolerance (54 min) was ketamine/ xylazine/acepromazine. Three further anesthetic combinations were associated with surgical tolerance: ketamine/ xylazine, ketamine/xylazinelazaperone, and tiletamine/xylazine/zolazepam (Telazol/xylazine). The protocols consisting of ketamine/medetomidine and ketamine/azaperone were not associated with clearly detectable surgical tolerance. The most reliable parameter of surgical tolerance under routine laboratory conditions was the pedal withdrawal reflex. CONCLUSIONS: The best intraperitoneal injection anesthesia regimen consisted of ketamine/xylazine/acepromazine. The dose must be adapted to the particulars of each experimental design (mouse strain, sex, age, mutation). This is best done by measuring surgical tolerance, using the pedal withdrawal reflex. PMID- 11924806 TI - Anesthesia protocol for hyperpnea-induced airway obstruction in the guinea pig. AB - Guinea pigs (GP; Cavia porcellus) are used extensively as an experimental animal model in a wide range of disciplines including respiratory physiology. Guinea pigs are difficult to anesthetize, and many investigators use paralytic agents to eliminate spontaneous respiratory movements; however, strict federal regulations and institutional policies governing use of paralytic agents are few. We report an anesthesia protocol, using the injectable anesthetic agents sodium pentobarbital (SP) and xylazine (XYL) for the GP that induces consistent anesthesia while eliminating use of paralytic agents. Sixty percent of the calculated SP dose (45 mg/kg of body weight) was given for anesthesia induction, followed by 50% of the calculated XYL dose (7 mg/kg) 15 min later. Depth of anesthesia was monitored by response to toe pinch, ECG, and spontaneous respiratory movements. The animals were given additional boosts of SP (5 to 15% of the original dose, i.p. or i.v.) if a change in anesthesia depth was noted. Thirty-one animals completed the hyperpnea-induced bronchoconstriction (HIB) study with no fatalities. Using this protocol, we collected consistent, repeatable, and reliable data without use of propranolol or skeletal muscle paralytics. We believe that this protocol is not restricted to the GP and could be adapted for use in other terminal experiments. PMID- 11924807 TI - Abnormalities of gonadotrophs in the adenohypophysis of infertile male PD rats. AB - Male PD strain rats are sterile in the homozygous condition (pd/pd) due to abnormal spermatogenesis detectable at around nine weeks of age. Previous studies have indicated electron microscopic abnormalities in the Sertoli cells of pd/pd males at three and 12 weeks of age. Since spermatogenesis and Sertoli cell function depend on gonadotropins (luteinizing and follicle-stimulating hormones [LH and FSH]) and testosterone, production and/or secretion of these hormones might be altered in pd/pd males. The aim of the study reported here was to investigate the hormonal status of pd/pd males at three, six, and nine weeks of age. Although alteration was not evident in the LH-immunoreactive cells, FSH immunoreactive cells in pd/pd males were small in size with scant cytoplasm and were reduced in number and area (73 and 51% of phenotypically normal pd/+ males, respectively) at three weeks of age, although serum FSH concentration was similar to that in pd/+ males. At six and nine weeks of age, percentages of the areas occupied by LH- and FSH-immunoreactive cells in pd/pd males were higher than those in pd/+ males. Serum FSH concentration in pd/pd males was significantly high at nine weeks of age, although a difference in serum LH and testosterone concentration was not evident. These results suggest that FSH production in pd/pd males is decreased at three weeks of age. This might be associated with the Sertoli cell abnormalities and subsequent abnormal spermatogenesis seen in adult life. PMID- 11924808 TI - Latex sensitivity in a macaque (Macaca mulatta). AB - BACKGROUND AND HISTORY: An adult Macaca mulatta was examined because of a history of multiple episodes of conjunctivitis and an acute, pruritic, dermatitic eruption that affected the axillary and inguinal regions, forearms, thorax, and neck. METHODS AND RESULTS: Results of corneal staining, examination of skin scrapings and feces, fungal culture, CBC, and a thyroid profile (thyroxine/triiodothyronine concentrations) were negative or normal, with the exception of eosinophilia (1,040/mm3). Examination of a punch biopsy specimen of the skin indicated chronic, nonsuppurative eosinophilic dermatitis. Skin patch testing against 25 contact allergens was negative for a delayed-type hypersensitivity reaction. Allergen-specific IgE testing, using six monkey chow additives, also yielded negative results, but testing against latex revealed a strong positive result (0.74 KU/L) consistent with a latex allergy. A skin prick test performed by use of a latex supernatant revealed significant inflammation at the latex site at 72 h and one week. Vinyl gloves were substituted for latex gloves, and that resulted in a marked decrease in erythema, pruritus, and lichenification with no flares of dermatitis for four years. Repeat skin biopsy fourteen weeks after the original biopsy revealed normal epidermis; however, mild chronic active nonsuppurative, perifolliculitis persisted. CONCLUSION: Latex can induce allergic dermatitis in nonhuman primates and should be included in the differen tial diagnosis for atopic dermatitis. PMID- 11924809 TI - Thinking inside and outside the box. PMID- 11924810 TI - Re: Laboratory animal medicine in a time of crisis. PMID- 11924811 TI - Anesthesia update: agents, definitions, and strategies. AB - The trend in modern anesthesia is to "lighten up." This generally involves use of several drugs with selective and complementary actions. The pharmacokinetic properties of such drugs should allow rapid onset, rapid recovery, and rapid responses to changes in delivered doses. Peri-operative management issues also are inherent to use of modern drugs and techniques. For example, provisions must be in place for postoperative analgesia if rapid recovery is anticipated. Light anesthesia reduces morbidity and mortality, and reduces the drug, facility, and personnel costs associated with anesthesia. However, the requirements for anesthesia and the expertise of personnel administering anesthesia vary considerably. Many regulatory bodies and scientific journals require a description of how anesthesia adequacy and depth will be assessed, as well as extensive justification for the use of neuromuscular blocking agents. In environments where adequate experience and sophistication for the use of cutting edge drugs and methods are not available, older drugs and techniques may be adequate and preferable to protect animals from pain or distress. PMID- 11924812 TI - Evaluation of the rat stifle joint after transection of the cranial cruciate ligament and partial medial meniscectomy. AB - Osteoarthritis (OA) was induced in the rat stifle joint by partial medial meniscectomy (PMM) and transection of the cranial cruciate ligament (CCL). At 10 weeks after destabilization, joint morphologic and pathologic changes were observed, scored, and compared. The intact rat stifle joint was observed in a mid saggital plane. Articular cartilage of the distal portion of the femur and proximal portion of the tibia had thicker and thinner sites, and the thicker sites were located caudally on the distal portion of the femur and centrally on the proximal portion of the tibia. The two separate triangular portions of the medial meniscus observed in the mid-saggital plane contained a center of ossification in the cranial portion and fibrocartilage in the caudal portion. The synovium was one to three cells thick, and contained rare inflammatory cells. Although lesions were more severe in stifles after PMM, both treatments produced OA lesions that closely simulated OA lesions of other species. Lesions consistent with idiopathic OA included chondrocytic clones with increased metachromasia around them, chondrocytic death, loss of metachromasia, fibrillation, fissuring, erosion of articular cartilage, osteophyte formation, and variable synovial inflammation. The results indicate that PMM and CCL transection in the rat are useful in vivo models for study of the etiopathogenesis of OA and therapeutic efficacy of anti-arthritic drugs and treatment concepts. PMID- 11924813 TI - Left ventricular, systemic arterial, and baroreflex responses to ketamine and TEE in chronically instrumented monkeys. AB - Effects of prescribed doses of ketamine five minutes after application and influences of transesophageal echocardiography (TEE) on left ventricular, systemic arterial, and baroreflex responses were investigated to test the hypothesis that ketamine and/or TEE probe insertion alter cardiovascular function. Seven rhesus monkeys were tested under each of four randomly selected experimental conditions: (1) intravenous bolus dose of ketamine (0.5 ml), (2) continuous infusion of ketamine (500 mg/kg/min), (3) continuous infusion of ketamine (500 mg/kg/min) with TEE, and (4) control (no ketamine or TEE). Monkeys were chronically instrumented with a high fidelity, dual-sensor micromanometer to measure left ventricular and aortic pressure and a transit-time ultrasound probe to measure aortic flow. These measures were used to calculate left ventricular function. A 4-element Windkessel lumped-parameter model was used to estimate total peripheral resistance and systemic arterial compliance. Baroreflex response was calculated as the change in R-R interval divided by the change in mean aortic pressure measured during administration of graded concentrations of nitroprusside. The results indicated that five minutes after ketamine application heart rate and left ventricular diastolic compliance decreased while TEE increased aortic systolic and diastolic pressure. We conclude that ketamine may be administered as either a bolus or continuous infusion without affecting cardiovascular function 5 minutes after application while the insertion of a TEE probe will increase aortic pressure. The results for both ketamine and TEE illustrate the classic "Hawthorne Effect," where the observed values are partly a function of the measurement process. Measures of aortic pressure, heart rate, and left ventricular diastolic pressure should be viewed as relative, as opposed to absolute, when organisms are sedated with ketamine or instrumented with a TEE probe. PMID- 11924814 TI - Reduction in animal numbers by long-term implantation of intravenous and intra arterial catheters in thyroparathyroidectomized rats. AB - The thyroparathyroidectomized (TPTx) rat has been extensively used to study parathyroid hormone (PTH)-mediated bone resorption by measuring systemic Ca2+ concentrations. Animals have been traditionally used acutely; that is, they are often infused immediately after surgery and are sacrificed after a single use. To perform multiple experiments using a single group of animals we developed a system of long-term implanted intravenous/arterial catheters. Using calcitonin (CT) as a positive control, we successfully completed 12 separate controlled subexperiments documenting significant reductions in PTH-induced hypercalcemia in rats of the CT group. We then successfully completed two separate TPTx subexperiments, using a 3 x 3 Latin square experimental design. In both subexperiments, CT significantly inhibited the increase of blood Ca2+ concentration resulting from continuous PTH infusion. Our results indicate that, by combining the long-term use of catheters with the Latin square design, we can successfully reduce the number of animals used, increase the number of compounds screened, and improve the quality of the data. Although results of this study confirmed the acceptability of multiple infusions in anti-resorptive studies, investigations into the applicability of this set up to other areas of study requiring infusions and frequent blood sample collections seem appropriate. PMID- 11924815 TI - Transcutaneous blood gas monitoring in the rat. AB - Transcutaneous blood gas (TCBG) analysis is a noninvasive alternative method of estimation of blood gas tensions. The objective of the study reported here was to validate this method against standard blood gas (STBG) analysis in adult and juvenile Sprague-Dawley rats. We sought to establish the optimal TCBG probe site and temperature, to establish probe temperatures that would not cause thermal burns, to evaluate correlations between blood gas values (PaCO2 and PaO2) determined by use of TCBG and STBG, and to evaluate the sensitivity of the TCBG unit to changes in arterial blood gas partial pressures. Our results indicated that: in general, the xyphoid area was the optimal site for probe placement, with 44.5 degrees C being the optimal probe temperature for the highest correlation, but thermal burns may be a problem; probe temperatures of 42.5 degrees C (adults) and 42.0 degrees C (juveniles) do not cause thermal burns when left in place for three hours; probe temperatures of 44 degrees C (adults) and 42 degrees C (juveniles) resulted in moderate correlation between PaCO2 and PtcCO2; and the TCBG unit adequately responded to changes in arterial blood gas partial pressures. Neither PtcCO2 or PtcO2 reflect actual values of PaCO2 or PaO2, respectively. We concluded that TCBG analysis may be used as an indicator of change in PaCO2 with sufficient animal numbers under tightly controlled conditions, but not as an indicator of change in PaO2 in adult and juvenile rats. PMID- 11924816 TI - Anatomic and physiologic reference values in least shrews (Cryptotis parva). AB - BACKGROUND AND PURPOSE: The least shrew is an established animal model for reproductive and pharmacologic research. Biologic reference data are needed to assess animal health status and provide a rationale for use of novel statistical programs to evaluate the effects of orally administered substances in toxicologic and pharmacologic studies. METHODS: Organ weights, blood biochemical and hematologic values, and food and water consumption data were collected from 50 day-old shrews after two weeks' consumption of a standard feline diet. RESULTS: In general, data correlated well with values reported for other mammalian species. Plasma phosphorus concentration was high. There was a significant difference in food and water consumption per gram of body weight between shrews at lower and upper (+/- 1 SD) weight ranges for the study. The 3.2-g animals consumed 27% more food per gram of body weight than did the 5.0-g animals. CONCLUSIONS: The high phosphorus concentration was attributed to hemolysis resulting from the axillary cut method of blood sample collection. The small size of the shrew allowed demonstration of the Kleiber effect within a +/- 1 SD weight range in a single species. The phenomenon necessitates the use of statistical methods other than the typical tests establishing the significance of the differences between the means of groups for oral toxicologic and pharmacologic studies. PMID- 11924817 TI - Mycobacterium avium subspecies paratuberculosis triggers intestinal pathophysiologic changes in beige/scid mice. AB - We investigated whether infection of beige/scid mice with Mycobacterium avium subspecies paratuberculosis can induce intestinal pathophysiologic changes. Six week-old beige/scid mice were inoculated intraperitoneally with M. paratuberculosis, then were killed 32 weeks after inoculation when the small intestine was evaluated for physiologic and morphologic abnormalities. All infected mice developed clinical disease. The lamina propria of the intestine from infected mice was mildly infiltrated with mononuclear cells containing acid fast bacteria, and had significantly increased villus width. In vitro physiologic studies in Ussing chambers indicated that M. paratuberculosis infection caused significant abnormalities in intestinal transport parameters. Baseline short circuit current and potential difference were abnormally high in tissues from infected, compared with control mice, indicative of increased ion secretion. Baseline conductance was significantly decreased in infected mice, suggesting that intestinal tissue from infected mice was less permeable to ions. The change in short circuit current following transmural electrical and glucose stimulation was significantly reduced in intestines from infected mice, suggesting that inflamed intestine had neural and/or epithelial cell damage. We conclude that infection of beige/scid mice with M. paratuberculosis triggers significant intestinal pathophysiologic changes consistent with chronic inflammation. These functional abnormalities may contribute to the pathogenesis of the wasting syndrome seen in bovids with paratuberculosis. This animal model provides evidence that T cell-independent mechanisms are sufficient to cause mucosal pathophysiologic changes and inflammation in response to a specific pathogen, and may be of relevance to inflammatory bowel disease in humans. PMID- 11924818 TI - Effects of birth season, breed, sex, and sire family on cardiac morphology determined in pigs (Sus scrofa domestica) by use of echocardiography. AB - PURPOSE: Echocardiography played an important role in the screening and diagnosis of hypertrophic cardiomyopathy. In the study reported here, we attempted to evaluate the effects of birth season, breed, sex, and sire family on cardiac morphology determined in pigs by use of echocardiography. METHODS: A total of 411 pigs (mean body weight and age of 105.7 +/- 10.6 kg and 214.4 +/- 25.5 days, respectively) with different genetic backgrounds (Landrace, Yorkshire, and their two-way crossbred) were studied. Cardiac morphologic measurements included thickness of left ventricle and interventricular septum at end-systolic and end diastolic phases. Meanwhile, the statistical model included the following effects: birth season, breed, sex, interaction between breed and sex, sire family, body weight, and age. RESULTS: Mean cardiac morphologic measurements were as follows: thickness of the interventricular septum at end-systolic and end diastolic phases was 1.74 and 1.14 cm, respectively; and thickness of the left ventricular free wall at end-systolic and end-diastolic phases was 1.81 and 0.98 cm, respectively. Medium positive correlations existed among the cardiac morphologic measurements r = 0.31 to 0.53; P < 0.001). Pigs born in spring had significantly (P < 0.05) lower cardiac thickness at the end systolic phase than did pigs born in other seasons, and Landrace pigs had higher cardiac morphologic measurements than did Yorkshire and two-way crossbred pigs. Additionally, thickness of interventricular septum at the end-diastolic phase in male pigs was significantly higher than that in female pigs (P < 0.05). Cardiac morphologic measurements for the sire family were significantly (P < 0.05) different, and contributed 77.2 to 87.9% of the total variation, suggesting that genetic variation in cardiac morphology might exist in pigs. CONCLUSIONS: Cardiac morphology of pigs might be influenced by genetic background. The effects of birth season, breed, sire family, and sex should be adjusted when using pigs as an animal model for comparative cardiovascular studies. PMID- 11924819 TI - Circling mouse: possible animal model for deafness. AB - Mutant mice with abnormalities are potentially useful as models for studying human defects. Here we report a group of mice with abnormal behavioral patterns. A new spontaneous mutant mouse exhibited hyperactive behavior at about seven days of age, followed by tight circling behavior. Breeding studies suggest that this mutation is caused by a single gene defect inherited in an autosomal recessive manner. Consequently, this mutation is referred to as a circling (cir) mouse mutation with the gene symbol cir. Auditory test results identified clearly the hearing loss of the cir, compared with wild-type mice. Pathologic studies confirmed developmental defects in cochlea and spiral ganglions that were correlated to the abnormal behavior observed in the cir mice. Thus, cir mice may be useful as a model for studying inner ear abnormalities and deafness in humans. PMID- 11924820 TI - Comparison of the frequencies of major histocompatibility (MHC) class-II DQA1 and DQB1 alleles in Indian and Chinese rhesus macaques (Macaca mulatta). AB - The major histocompatibility complex (MHC) comprises related gene families, some of which are highly polymorphic, whose protein products mediate immune response. Rhesus macaques (Macaca mulatta) are a vital animal model for research in human diseases and are native to regions extending from Afghanistan in the west to the Eastern Plains of China and from Peking to the north, southward through islands of Southeast Asia. The distributions of MHC class-II Mamu DQA1 and Mamu DQB1 alleles in two groups of domestically bred rhesus macaques of Indian and Chinese origin and the Mamu DQA1 genotypes of a small number of Burmese rhesus macaques were compared. Major allelic differences were observed between the Indian and Chinese rhesus macaques, and gene diversity decreased from east to west. These and other intra-specific genetic differences among regional populations of rhesus macaques might influence the outcome of biomedical research in which they are used as subjects, and illustrate the importance of completely genetically characterizing subjects used as animal models in biomedical research. PMID- 11924821 TI - Anisocoria and middle cerebral artery saccular (berry) aneurysm in a rhesus macaque (Macaca mulatta). AB - A 27-year-old female rhesus macaque (Macaca mulatta) developed anisocoria. The left pupil was dilated and unresponsive to light. The macaque was euthanized because of unrelated reasons and the body was submitted for necropsy. On gross examination, a berry aneurysm of the right middle cerebral artery causing marked compression of the right optic tract was found. Arteriosclerotic changes were observed microscopically in the right middle cerebral and in the internal carotid arteries. The left iris was markedly degenerated, with atrophy of the constrictor muscle. Compression of the right optic tract may cause homonimus hemianopsia. A dilated and unresponsive left pupil indicated a lesion in the ipsilateral parasympathetic efferent pathway. In the absence of appreciable lesions of the left oculomotor nerve, the most likely cause of mydriasis was the iridic lesion. Intracranial aneurysms are common in humans (2 to 5%), but not in other species. Only about 10% of unruptured aneurysms are associated with neurologic deficits related to mechanical compression, such as visual deficits or anisocoria. Meticulous investigation of the ocular vascular and neural pathways led us to conclude that the anisocoria was unrelated to the aneurysm. To our knowledge, this report represents the first documented case of a naturally occurring intracranial aneurysm in nonhuman primates. PMID- 11924822 TI - Microbial ecology of human skin in health and disease. AB - Cultivation of human skin reveals numerous bacteria and at least one fungus to be normal inhabitants of this ecosystem; however, most of our knowledge about the microbiology of human skin was acquired decades ago. Modern techniques employing nucleic acid-based microbial identification methods demonstrate the limitations of cultivation for appreciating microbial diversity in many ecosystems. The application of modern molecular methods to the study of skin may offer new perspectives on the resident microfora, and new insights into the causes of antibiotic responsive dermatologic conditions, such as acne and rosacea. PMID- 11924823 TI - Skin microflora and bacterial infections of the skin. AB - The skin is a milieu for controlled bacterial growth. Skin supports the growth of commensal bacteria, which protect the host from pathogenic bacteria. Environmental and local factors, host immunity, and organism adherence and virulence are intricately related to cutaneous infection. Resident gram-positive bacteria include Staphylococcus, Micrococcus, and Corynebacterium sp. Staphylococcus aureus and Streptococcus pyogenes are notoriously pathogenic in the skin. In order for bacteria to be pathogenic, they must be able to adhere to, grow on, and invade the host. Bacteria possess numerous virulence genes that allow for growth in these privileged niches. Epidermal infections caused by S. aureus and S. pyogenes include impetigo and ecthyma. Dermal infections consist of erysipelas, cellulitis, and necrotizing fasciitis. The pilosebaceous unit is involved in folliculitis, furunculosis, and carbunculosis. Moreover, S. aureus and S. pyogenes produce toxins that may elicit a superantigen response, causing massive release of cytokines. Staphylococcal scalded skin syndrome, toxic shock syndrome, and scarlet fever are all superantigen-mediated. Gram-negative organisms such as Pseudomonas aeruginosa, Pasteurella multocida, Capnocytophaga canimorsus, Bartonella sp., Klebsiella rhinoscleromatis, and Vibrio vulnificus are not typical resident skin microflora but may cause cutaneous infection. PMID- 11924824 TI - Leishmaniasis as an emerging infection. AB - Leishmaniasis is a protozoan disease whose diverse clinical manifestations are dependent both on the infecting species of Leishmania and the immune response of the host. Transmission of the disease occurs by the bite of a sand fly infected with Leishmania parasites. Infection may be restricted to the skin in cutaneous leishmaniasis, limited to the mucous membranes in mucosal leishmaniasis, or spread internally in visceral leishmaniasis or kala azar. The overall prevalence of leishmaniasis is 12 million cases worldwide, and the global yearly incidence of all clinical forms approaches 2 million new cases (World Health Organization WHO/ LEISH/200.42, Leishmania/HIV Co-Infection in Southwestern Europe 1990-98: Retrospective Analysis of 965 Cases, 2000). In the last two decades, leishmaniasis, especially visceral leishmaniasis, has been recognized as an opportunistic disease in the immunocompromised, particularly in patients infected with human immunodeficiency virus. PMID- 11924825 TI - Varicella-zoster virus: a re-emerging infection. AB - Varicella-zoster virus (VZV) causes chickenpox (varicella), becomes latent in cranial nerve and dorsal root ganglia, and can reactivate many years later to produce shingles (zoster) and postherpetic neuralgia (PHN). Elderly and immunocompromised individuals are also at risk for complications of VZV reactivation involving the central nervous system (CNS), including myelitis, large-vessel encephalitis/granulomatous arteritis, small-vessel encephalitis, meningoencephalitis, and ventriculitis. Peripheral nervous system (PNS) complications range from zoster and postherpetic neuralgia to postinfectious polyneuritis (Guillain-Barre syndrome, GBS). These complications can occur with or without cutaneous manifestations. An increase in elderly and immunocompromised individuals will likely result in a higher prevalence of these conditions; therefore, VZV can be seen as a "re-emerging" infection of the early twenty-first century. In this review, we summarize our experience and the existing literature on CNS and PNS complications of VZV reactivation. Special attention is paid to reports of complications without rash, as these entities are more difficult to diagnose. PMID- 11924826 TI - Candida: a causative agent of an emerging infection. AB - Incidences of infections due to Candida have increased over the last 15-20 y. This increase in the incidence and the high associated mortality rate despite therapy has focused the attention on this disease and prompted investigators to undertake research aimed at understanding the pathogenesis of this disease as well as methods to treat it. This paper discusses recent developments in the Candida field and the impact they have on patient management. PMID- 11924827 TI - Skin diseases associated with human herpesvirus 6, 7, and 8 infection. AB - Relatively recently, the discovery and analysis of three new human herpesviruses, human herpesvirus (HHV)-6, HHV-7, and Kaposi's sarcoma-associated herpesvirus (KSHV), also known as HHV-8, has contributed greatly to our understanding of the pathogenesis of several common dermatoses. HHV-6 and HHV-7 are closely related beta-herpesviruses that have been linked with roseola (mostly HHV-6), severe drug eruptions (HHV-6), and pityriasis rosea (mostly HHV-7). KSHV is a gamma herpesvirus that is now believed to be the long sought after etiologic agent of Kaposi's sarcoma. The evidence for these skin disease associations and key findings from recent basic science investigations on viral pathogenesis are discussed in this review. In addition, possible therapeutic implications of these research studies are explored. PMID- 11924828 TI - Human papillomavirus and skin cancer. AB - Human papillomavirus (HPV) appears to be the most ubiquitous of the human viruses. Over 100 HPV types have been identified. A minority of HPV cause cutaneous warts and mucosal condylomata. The HPV that cause mucosal condylomata put the patient at various degrees of risk for developing cancers, particularly cervical cancer. The majority of HPV infect the skin of normal and immunocompromised individuals. In normal people, most of these HPV appear to establish a latent infection of the skin, most likely as normal flora residing in hair follicles; however, in patients with various systemic and localized depressions of cell-mediated immunity, some HPV infections appear to be involved in the development of nonmelanotic skin cancer and its precursor lesions in skin, usually in sunlight-exposed areas. Circumstantial evidence suggests that these HPV may have a role in promoting proliferative lesions of the skin, although their sites of active infection and mode of transmission to susceptible individuals remain unknown. PMID- 11924829 TI - Human papillomavirus infection and skin cancer risk in organ transplant recipients. AB - Warts and squamous cell carcinomas are important cutaneous complications in organ transplant recipients. The role of infection with human papillomaviruses (HPV) in the development of cutaneous squamous cell carcinoma is still unclear. An extremely diverse group of HPV types, mainly consisting of epidermodysplasia verruciformis (EV)-associated HPV types, can be detected in benign, premalignant, and malignant skin lesions of organ transplant recipients. Frequently, there are multiple HPV types present in single skin biopsies. Typically, the prevalence of viral warts rises steadily after transplantation and a strong association exists between the number of HPV-induced warts and the development of skin cancer. The interval between the transplantation to the development of warts is clearly shorter than the interval from transplantation to the diagnosis of the first skin cancer. A comparison of transplant recipients with and without skin cancer, however, showed an equally high prevalence of EV-HPV DNA in keratotic skin lesions in both groups of patients and the detection rate and spectrum of HPV infection in hyperkeratotic papillomas, actinic keratoses, and squamous cell carcinomas was also similar. HPV DNA can frequently be detected in patients with hyperproliferative disorders like psoriasis and antibodies against HPV in patients with regenerating skin (e.g., after extensive second degree burns). Latent infection with EV-HPV seems to be widespread. The hair follicle region might be the reservoir of EV-HPV. The E6 protein from a range of cutaneous HPV types effectively inhibits apoptosis in response to UV-light induced damage. It is therefore conceivable that individuals who are infected by EV-HPV are at an increased risk of developing actinic keratoses and squamous cell carcinomas, possibly by chronically preventing UV-light induced apoptosis. PMID- 11924830 TI - Immune reconstitution in HIV-1-infected individuals treated with potent antiretroviral therapy. AB - Potent combination antiretroviral therapy that was introduced in the mid-1990s for treatment of HIV-1 infection has resulted in unprecedented decreases in HIV-1 replication and increases in CD4+ T cell counts in many individuals. Coincident with the introduction of potent combination antiretroviral therapy, substantial declines in AIDS-related morbidity and mortality have been observed. Although these declines strongly suggest that significant immune reconstitution is occurring, increasing evidence suggests that immune reconstitution is neither uniform nor complete in all treated individuals. Clinical data suggest that some HIV-1-associated malignancies have not declined despite the new therapies, and that not all treated individuals reconstitute CD4+ T cell numbers to normal values. Laboratory studies reveal that immune responses to ubiquitous antigens are reconstituted, but that responses to rarely encountered antigens, such as tetanus, are not reconstituted without repeat vaccination. Many questions remain concerning the extent and clinical significance of the immune reconstitution that occurs in the setting of antiretroviral drug therapy. A better understanding of the nature of the immune reconstitution that results from potent antiretroviral therapy is critical to the optimal clinical management of HIV-1-infected individuals, and may provide important insights into the immunopathogenesis of HIV-1 infection as well. PMID- 11924831 TI - Human dendritic cells as targets of dengue virus infection. AB - Dengue virus infections are an emerging global threat. Severe dengue infection is manifested as dengue hemorrhagic fever and dengue shock syndrome, both of which can be fatal complications. Factors predisposing to complicated disease and pathogenesis of severe infections are discussed. Using immunohistochemistry, immunofluorescence, flow cytometry, and ELISA techniques, we studied the cellular targets of dengue virus infection, at both the clinical (in vivo) and the laboratory (in vitro) level. Resident skin dendritic cells are targets of dengue virus infection as demonstrated in a skin biopsy from a dengue vaccine recipient. We show that factors influencing infection of monocytes/macrophages and dendritic cells are different. Immature dendritic cells were found to be the cells most permissive for dengue infection and maybe early targets for infection. Immature dendritic cells exposed to dengue virus produce TNF-alpha protein. Some of these immature dendritic cells undergo TNF-alpha mediated maturation as a consequence of exposure to the dengue virus. PMID- 11924832 TI - The keratinocyte as a target for staphylococcal bacterial toxins. AB - Skin infections with Staphylococcus aureus are not only an important cause of morbidity and even mortality, but are thought to serve as initiation and/or persistance factors for numerous inflammatory skin diseases, including psoriasis and atopic dermatitis. One mechanism by which S. aureus can modulate the immune system is through the production of proteins such as superantigenic toxins, Protein A, as well through the cytolytic alpha-toxin. This review serves to discuss the biology of these three types of proteins, with emphasis on their ability to stimulate the production of powerful pro-inflammatory lipid- and protein-derived cytokines in keratinocytes. Characterization of interactions between these proteins and the keratinocyte can provide a better understanding of how bacterial infection modulates inflammatory skin diseases, as well as provide the basis for improved therapies involving antibacterial agents. PMID- 11924833 TI - Biologic effects of bacterial superantigens in a xenogeneic transplantation model for psoriasis. AB - Both clinical as well as experimental data support the concept of psoriasis being a T-cell-mediated immune disease possibly triggered by bacterial superantigens. Further analysis of its pathogenesis was facilitated by the generation of a xenogeneic transplantation model in which skin from psoriatic patients is grafted onto SCID mice lacking functional B and T cells. Applying this model it was demonstrated that psoriasis can be triggered by bacterial superantigens; this process depends on the presence of immunocytes. Mutated variants of the respective superantigens exhibiting no measurable affinity to HLA class II molecules can function as competitive inhibitors in vivo. PMID- 11924834 TI - Chlamydia pneumoniae and chronic skin wounds: a focused review. AB - The genus, Chlamydophilia, as obligate intracellular pathogens, induce chronic scarring in humans. Chlamydia pneumoniae, a common cause of pneumonia, infects endothelial cells and circulating macrophages. Evidence that C. pneumoniae is an opportunistic pathogen in chronic skin ulcers and other inflammatory skin conditions analogous to its role in atherosclerosis is reviewed. PMID- 11924835 TI - Prophylactic and therapeutic vaccines for genital papillomavirus infection. AB - The development of potential therapeutic and prophylactic vaccines for human papillomavirus (HPV) infection is a very exciting area of HPV research. There are a number of features of HPV biology that makes the development of a vaccine particularly difficult, although there are several examples of vaccines that have had spectacular success in the prevention of other viral diseases. Our poor understanding of the immune response to HPV infection is the first problem. We do not understand the mechanism by which spontaneous clearing of warts is generated and therefore cannot particularly target this pathway in the development of a vaccine. Furthermore, there is no in vitro culture system nor an animal model for HPV. Another problem is that there is no ready source of live virus that might be exploited for a live attenuated viral vaccine, such as was used with poliovirus. Although most other viruses spend a portion of their life cycle in the systemic circulation where they are vulnerable to neutralizing antibodies, HPV remain exclusively in the epithelium and thus antibodies must transverse the basement membrane and reach the other layers of the skin or mucosa to be effective in preventing infection. Significant progress is being made in the development of potential vaccine candidates despite these and other confounding factors. PMID- 11924836 TI - Anti-cytokine therapies in response to systemic infection. AB - In the past 5 y, the 28 d mortality in patients with sepsis syndrome has decreased somewhat but still ranges from 30% to 40%; mortality in those patients with septic shock and multiple organ failure is higher. This high mortality is observed despite intensive care units that deliver hemodynamic, metabolic, ventilatory, and renal support. Clearly some patients survive the ordeal but it remains frustrating not being able to stop the downhill course leading to multiple organ failure and death in these patients. New therapies have been sought and tested, including those preventing the biologic activity of two pro inflammatory cytokines, interleukin-1 (IL-1) and tumor necrosis factor (TNF). Based on animal studies, anti-TNF and IL-1 therapy has been used to "rescue" the patient who continues to deteriorate in the face of considerable support efforts. Unfortunately, these anticytokine therapies have not dramatically reduced 28 d mortality in double-blind, placebo-controlled trials involving nearly 10000 patients, although there is a consistent but statistically nonsignificant decrease in mortality associated with anticytokine therapies. On the other hand, the same anti-TNF and IL-1-based therapies have made a dramatic improvement in the local inflammation and progression of rheumatoid arthritis. It appears that systemic inflammation of sepsis requires more than anticytokine monotherapy to significantly reduce mortality. PMID- 11924837 TI - The cross-modal coordination of interpersonal timing: six-week-olds infants' gaze with adults' vocal behavior. AB - The purpose of this study was to examine the hypothesis that 6-week-old infants are capable of coordinated interpersonal timing within social interactions. Coordinated interpersonal timing refers to changes in the timing of one individual's behavior as a function of the timing of another individual's behavior. Each of 45, first-bom 6-week-old infants interacted with his or her mother and a stranger for a total of 14 minutes. The interactions were videotaped and coded for the gaze behavior of the infants and the vocal behavior of the mothers and strangers. Time-series regression analyses were used to assess the extent to which the timing of each of the infants' gazes was coordinated with the timing of the adults' vocal behavior. The results revealed that (a) coordinated timing occurs between infants and their mothers and between infants and strangers as early as when infants are 6 weeks old, and (b) strangers coordinated the timing of their pauses with the infants to a greater extent than did mothers. The findings are discussed in terms of the role of temporal sensitivity in social interaction. PMID- 11924838 TI - Readers' eye movements distinguish anomalies of form and content. AB - Evidence is presented that eye-movement patterns during reading distinguish costs associated with the syntactic processing of sentences from costs associated with relating sentence meaning to real world probabilities. Participants (N = 30) read matching sets of sentences that differed by a single word, making the sentence syntactically anomalous (but understandable), pragmatically anomalous, or non anomalous. Syntactic and pragmatic anomaly each caused perturbations in eye movements. Subsequent to the anomaly, the patterns diverged. Syntactic anomaly generated many regressions initially, with rapid return to baseline. Pragmatic anomaly resulted in lengthened reading times, followed by a gradual increase in regressions that reached a maximum at the end of the sentence. Evidence of rapid sensitivity to pragmatic information supports the use of timing data in resolving the debate over the autonomy of linguistic processing. The divergent patterns of eye movements support indications from neurocognitive studies of a principled distinction between syntactic and pragmatic processing procedures within the language processing mechanism. PMID- 11924839 TI - Distinct neurophysiological patterns reflecting aspects of syntactic complexity and syntactic repair. AB - Aspects of syntactic complexity and syntactic repair were investigated by comparing the event-related (brain) potentials (ERPs) for sentences of different syntactic complexity to those containing a syntactic violation. Previous research had shown that both aspects of syntactic processing are reflected in a late positivity (P600). Results from the present reading experiment demonstrate, however, that although both processing aspects elicit a late positivity, they are different in distribution. The repair-related positivity preceded by a negativity displayed a centroparietal distribution, whereas the complexity-related positivity showed a frontocentral scalp distribution. These data indicate that the P600 is not a unitary phenomenon. Moreover, the distributional differences strongly suggest that different neural structures underlie the two aspects of processing, namely syntactic repair and syntactic integration difficulties, most evident when processing syntactically complex sentences. PMID- 11924840 TI - Comprehending noun phrase arguments and adjuncts. AB - Two experiments investigated how readers comprehend noun phrase (NP) arguments and adjuncts. Previous research suggested that argument phrases are processed more quickly than adjunct phrases (Clifton, Speer, & Abney, 1991; Kennison, 1999; Schutze & Gibson, 1999; Speer & Clifton, 1998). The present experiments investigated whether the type of verb in the sentence context could influence how NP arguments and adjuncts were processed. Reading time was measured on sentences containing NP arguments and adjuncts preceded either by verbs occurring most frequently with NP arguments (biased transitive verbs) or by verbs occurring most frequently without NP arguments (biased intransitive verbs) (e.g., "Meredith read/performed every play/week."). In Experiment 1, reading time was measured using a self-paced phrase-by-phrase moving window. In Experiment 2, reading time was measured using eye tracking. The results of both experiments indicated that, following biased transitive verbs, NP arguments were processed more quickly than NP adjuncts. When NPsfollowed biased intransitive verbs, there was no significant difference between the processing time of NP arguments and adjuncts. PMID- 11924841 TI - The time relationships of the constituent components of the human electrocardiogram. AB - This paper reports the results of an examination of the timing relationships of the principal constituent components of the human electrocardiogram (ECG). ECG recordings were obtained from 21 healthy subjects, 10 male and 11 female aged between 13 and 65 years, over a wide range of heart rates extending from 46 to 184 beats per minute (bpm). A wavelet transform method based on the Mexican Hat wavelet was then used to precisely locate the positions of the onset, peak, termination and the duration of individual components in the ECG. Component times were then classified according to the heart rate associated with the cardiac cycle to which the component belonged. Second-order equations in the square root of the cardiac cycle time, TR-R of theform AT(1/2)R-R+BT R-R+C were fitted to the data obtained for each component to characterize its timing variation. These equations may be used to synthesize an ECG signal having a profile that varies with heart rate in a manner which reflects the in vivo variation. PMID- 11924842 TI - Preliminary study of potential for rapid prototype and surface scanned radiotherapy facemask production technique. AB - Preliminary investigations into the potential of an elegant technique to create a rudimentary facemask for patient immobilization during radiotherapy treatment are presented. This method combines modern technology to cause the patientfar less discomfort compared with current plaster of Paris (POP) face mould procedures. Near instantaneous patient face scanning is accomplished with charge-coupled devices for imaging in an optical surface scanning system. The surface generated data is input to a rapid prototype (RP) system that creates a life-size model of the patient's face topology. As proof of principle a basic prototype facemask was successfully constructed using this technique and some qualitative comparison measurements for position and surface dose were made. These initial results confirm the validity of this technique and justify the need for further quantitative studies to fully investigate the potential of RP facemasks over POP based methods for mask production. PMID- 11924844 TI - Force assessment of the stimulated arm flexors: quantification of contractile properties. AB - The primary objective was to develop equipment and evaluate protocols for non invasive assessment of contractile properties of human arm flexors. The research design consisted of a non-randomized control trial, with repeated measures. Data from six males and two females were gathered in a clinical research laboratory. The elbow flexor torque following motor point or direct nerve stimulation was measured in response to single pulses or short trains of electrical pulses. Length--tension relationships were determined; comparative data were obtained at the identified optimal muscle lengths. Twitch waveforms and peak torques following either type of stimulation were reproducible (within 10%). Peak torques following a 4-pulse small interpulse interval stimulation were nearly identical for motor-point activation and direct nerve stimulation (15.2+/- 6.6Nm for motor point stimulation; 14.5 +/- 6.6Nm for nerve stimulation). Average perceived pain indexes associated with 4-pulse stimuli were slightly higher following nerve stimulation (782 for nerve versus 6.23 for motor-point, n=8). A reliable methodology (motor-point stimulation) has been identified to perform stimulated force assessment of human arm flexors. PMID- 11924843 TI - Electrical current distribution under transthoracic defibrillation and pacing electrodes. AB - The known effect of high current density under the perimeter of defibrillation electrodes, leading to skin damage and even severe burns in some cases, has been considered by many investigators. Two main approaches for improvement were proposed: (i) interfacing with layers of varying and high resistivity and (ii) lengthening and shaping the perimeter line. Using finite element and physical modelling, it is shown that the second approach does not yield significant improvement in the distribution uniformity. Moreover, the application of high resistivity layers is unacceptable in dibrillation. The use of a low resistance layer with a diameter covering and extending over the metal plate by at least 2.5 mm results in better uniformity. A similar effect can be obtained by recessing the metal plate in an isolating support--an approach adopted from implantable neurostimulation electrodes. These two versions can be applied in combination. PMID- 11924846 TI - Differences in foetal heart rate variability from phonocardiography and abdominal electrocardiography. AB - The agreement of the phonocardiographic method to provide foetal heart rate variability (FHRV) indices equivalent to those derived from abdominal electrocardiography was tested. 15 pregnant women were recruited in order to obtain antepartum foetal phonocardiograms and abdominal electrocardiograms three minutes long. From the respective sound (SS) and electric (RR) time series, typical temporal and spectral indices of FHRV were computed and compared. Means of the SS and RR intervals were not significant (P> 0.05) and these showed a correlation r=0.98. However, the temporal indices, P(TOT), HF and LF/HF presented differences (P< 0.05), since SS values were higher. Spectral coherence decreased below 0.5 for frequencies above 0.28+ 0.07 Hz, where bands resembling maternal and foetal breathing movements were noted. Particularly above 0.28Hz, temporal and spectral FHRV indices derived from phonocardiography and electrocardiography show differences. Quality of the signal, processing techniques, and maternal and foetal respiratory factors could account to explain these. PMID- 11924845 TI - Permanent magnetic-levitation of rotating impeller: a decisive breakthrough in the centrifugal pump. AB - Magnetic bearings have no mechanical contact between the rotor and stator, and a rotary pump with magnetic bearings therefore has no mechanical wear and thrombosis. The magnetic bearings available, however, contain electromagnets, are complicated to control and have high energy consumption. Therefore, it is difficult to apply an electromagnetic bearing to a rotary pump without disturbing its simplicity, reliability and ability to be implanted. The authors have developed a levitated impeller pump using only permanent magnets. The rotor is supported by permanent radial magnetic forces. The impeller is fixed on one side of the rotor; on the other side the rotor magnets are mounted. Opposite these rotor magents, a driving magnet is fastened to the motor axis. Thereafter, the motor drives the rotor via magnetic coupling. In laboratory tests with saline, where the rotor is still or rotates at under 4,000 rpm, the rotor magnets have one point in contact axially with a spacer between the rotor magnets and the driving magnets. The contacting point is located in the center of the rotor. As the rotating speed increases gradually to more than 4000 rpm, the rotor will disaffiliate from the stator axially, and become fully levitated. Since the axial levitation is produced by hydraulic force and the rotor magnets have a giro effect, the rotor rotates very stably during levitation. As a left ventricular assist device, the pump works in a rotating speed range of 5,000-8,000 rpm, and the levitation of the impeller is assured by use of the pump. The permanent maglev impeller pump retains the advantages of the rotary pump but overcomes the disadvantages of the leviated pump with electromagnetic-bearing, and has met with most requirements of artificial heart blood pumps, thus promising to have more applications than previously. PMID- 11924847 TI - Improved ventilation with CFD. PMID- 11924848 TI - QRS detection using new wavelets. AB - This paper deals with a new wavelet (WVT) which has been developed and very effectively and efficiently used for the detection of QRS segments from the ECG signal. After carrying out the detection using five existing wavelets (two symmetric--WT1 and WT2--and three asymmetric--WT3, WT4 and WT5), two new wavelets (WT6 and WT7) were constructed and used for QRS detection. WT6 is a symmetric wavelet and has been constructed by a trial-and-error method. WT7 is an adaptive symmetric wavelet and adjusts its threshold as per the amplitude of the ECG signal. The accuracy of QRS detection obtained from WT6 is 99.8 % and from WT7 100%. The CSE DS-3 database has been used for tests. Both WT6 and WT7 have been proved to be superior in performance to the existing wavelets. Out of WT6 and WT7, WT7 holds high promise for error-free reliable QRS detection in computer aided feature extraction and disease diagnostics. PMID- 11924850 TI - C3 or C4 macrophytes: a specific carbon source for the development of semi aquatic and terrestrial arthropods in central Amazonian river-floodplains according to delta13C values. AB - C4 plant species were proposed to generally represent inferior food sources compared to C3 plants thus are avoided by herbivores, particularly insects. This was tested in semi-aquatic and terrestrial arthropods from Amazonian river floodplains by carbon isotope discrimination (delta13C). Two semi-aquatic grasshopper species (Stenacris f. fissicauda, Tucavaca gracilis-Acrididae) obtain their carbon during development from specific C4 macrophytes and two semi-aquatic species (Cornops aquaticum-Acrididae, Paulinia acuminata-Pauliniidae) from specific C3 macrophytes. The terrestrial millipede Mestosoma hylaeicum (Paradoxosomatidae) obtains about 45% of its carbon from roots of one C4 macrophyte during the development of immatures whereas adults use other food sources, including C3 trees. Results suggest, that (1) both C4 and C3 plants represent distinct hosts for terrestrial arthropods in Amazonia; (2) immatures may use plant species with a different photosynthetic pathway than adults. PMID- 11924849 TI - Food web implications of delta13C and delta15N variability over 370 km of the regulated Colorado River USA. AB - Dual stable isotope analysis in the regulated Colorado River through Grand Canyon National Park, USA, revealed a food web that varied spatially through this arid biome. Down-river enrichment of delta13C data was detected across three trophic levels resulting in shifted food webs. Humpack chub delta13C and delta15N values from muscle plugs and fin clips did not differ significantly. Humpback chub and rainbow trout trophic position is positively correlated with standard length indicating an increase in piscivory by larger fishes. Recovery of the aquatic community from impoundment by Glen Canyon Dam and collecting refinements for stable isotope analysis within large rivers are discussed. PMID- 11924851 TI - Stable isotopes in animal ecology: the effect of ration size on the trophic shift of C and N isotopes between feed and carcass. AB - The feeding ecology of living or extinct animal species is now frequently studied by analysis of stable isotope ratios in small quantities of carcass or remains such as bones or teeth. Although there are numerous papers on these applications in natural systems, the theoretical and experimental basis of this method is weak. In order to evaluate the effect of different feeding levels on the carbon and nitrogen trophic shift, an experiment was carried out in which fish (Nile tilapia, Oreochromis niloticus, initial weight 40.8 g) were fed for 4 weeks at three levels from slightly above maintenance to almost satiation. For each treatment, three fish were reared individually. The isotopic ratios of carbon and nitrogen in feeds and fish carcasses were determined and in the case of carbon this was done separately for lipids and lipid-free matter. The trophic shift was calculated at each feeding level from the delta13C and delta15N ratios of feed and fish. There was a significant trend towards higher values for the trophic shift at higher feeding rates in all fractions analysed. Although further research is required, it can be concluded that the effect of feeding level cannot be ignored when the diet of an animal has to be calculated from the ratios of isotopes in its body. PMID- 11924852 TI - The measurement of muscle protein synthesis in broilers with a flooding dose technique: use of 15N-labelled phenylalanine, GC-MS and GC-C-IRMS. AB - An experiment was carried out to measure fractional muscle protein synthesis rates (k(s)) in broilers with injection of a flooding dose of phenylalanine (1 ml/100 g body weight of 150 mM phenylalanine; 38 atom percent excess (APE) [15N]phenylalanine). K(s) was calculated from the [15N] enrichment in phenylalanine of tissue-free and protein-bound phenylalanine using both gas chromatography mass spectrometry (GC-MS) and gas chromatography combustion isotope ratio mass spectrometry (GC-C-IRMS) for measurements after a 10 min isotope incorporation period. The tertiary-butyldimethylsilyl (t-BDMS) derivatives of phenylalanine were used for gas chromatographic separation in both systems. GC-MS and GC-C-IRMS were calibrated for a range of 7 to 37 [15N]APE and 0 to 0.62 [15N]APE, respectively, and for sample sizes of 0.45 to 4.5 nmol phenylalanine and 7 to 40 nmol phenylalanine, respectively. Reproducibility of standards as a measure of precision varied from 0.06 to 0.29 [15N]APE and from 0.0004 to 0.0018 [15N]APE in GC-MS and GC-C-IRMS, respectively. K(s) was measured in the m. pectoralis major of broilers fed rye based diets (56%) which were provided either unsupplemented (-) or supplemented (+) with an enzyme preparation containing xylanase. K(s) in breast muscles was significantly increased from 21.8%/d to 23.9%/d due to enzyme supplementation. It can be concluded from the study that the measurement of protein synthesis in broilers with the flooding dose technique can be carried out by using [15N]phenylalanine, GC-MS and GC-C IRMS. PMID- 11924853 TI - Metabolic effects of HAY's diet. AB - The aim of the study was, to evaluate the metabolic effect of HAY's diet on protein turnover, fat oxidation, respiratory quotient, body fat and weight loss. Twelve healthy adults received an individually regular diet and thereafter a corresponding isocaloric and isonitrogenous 10-day HAY-diet. Protein turnover and 13C-fat oxidation were investigated after administration of [15N]glycine and an [U-13C]algae lipid mixture. The 15N and 13C enrichment in urine and breath were measured by isotope ratio mass spectrometry. The respiratory quotient was measured by indirect calorimetry. Body fat, total body water and lean body mass were estimated by bio-electric impedance analysis. HAY's diet led to a significantly higher 13C-fat oxidation (15.4 vs. 22.0% P < 0.01), corresponding to a lower respiratory quotient (0.88 vs. 0.81; P < 0.01), whereas the protein turnover remained constant in both diets (3.06 vs. 3.05 g/kg/day). HAY's diet did not reduce total body water, lean body mass, body fat and body weight (72.2 vs. 71.4 kg). PMID- 11924854 TI - Carbon isotope fractionation in the decarboxylation of phenylpropiolic acid in hydrogen donating media. AB - 13C kinetic isotope effect (KIE) in the decarboxylation of phenylpropiolic acid (PPA) in tetralin medium (Tn) has been determined at 409-432 K and found to be of magnitude similar to the 13C KIE observed in the decarboxylation of malonic acid where the rupture of the C-C bond is the rate determining step. 13C KIE equals 1.0318/at 136 degrees C in the decarboxylation of PPA in Tn medium. Intramolecular 13C KIE in the decarboxylation of malonic acid equals 1.0316 at this temperature. Thus it has been shown that the nearly "full" 13C KIE can be achieved by providing the excess hydrogen to Calpha of PPA (or to triple acetylene bond) using not only strong mineral acids as the source of protons but also by carrying out the decarboxylation in organic medium like tetralin. A mechanism of decarboxylation of PPA in Tn is suggested. PMID- 11924855 TI - Systematic and regularities in the origin of 2H patterns in natural compounds. PMID- 11924856 TI - The nonlinear response of sulfate to a decrease in SO2. PMID- 11924857 TI - Secondary organic aerosol formation from the irradiation of simulated automobile exhaust. AB - A laboratory study was conducted to evaluate the potential for secondary organic aerosol formation from emissions from automotive exhaust. The goal was to determine to what extent photochemical oxidation products of these hydrocarbons contribute to secondary organic aerosol (SOA) and how well their formation is described by recently developed models for SOA formation. The quality of a surrogate was tested by comparing its reactivity with that from irradiations of authentic automobile exhaust. Experiments for secondary particle formation using the surrogate were conducted in a fixed volume reactor operated in a dynamic mode. The mass concentration of the aerosol was determined from measurements of organic carbon collected on quartz filters and was corrected for the presence of hydrogen, nitrogen, and oxygen atoms in the organic species. A functional group analysis of the aerosol made by Fourier transform infrared (FTIR) spectroscopy indicated PMID- 11924858 TI - Simultaneous control of Hg0, SO2, and NOx by novel oxidized calcium-based sorbents. AB - Efforts to develop multipollutant control strategies have demonstrated that adding certain oxidants to different classes of Ca-based sorbents leads to a significant improvement in elemental Hg vapor (Hg0), SO2, and NOx removal from simulated flue gases. In the study presented here, two classes of Ca-based sorbents (hydrated limes and silicate compounds) were investigated. A number of oxidizing additives at different concentrations were used in the Ca-based sorbent production process. The Hg0, SO2, and NOx capture capacities of these oxidant enriched sorbents were evaluated and compared to those of a commercially available activated carbon in bench-scale, fixed-bed, and fluid-bed systems. Calcium-based sorbents prepared with two oxidants, designated C and M, exhibited Hg0 sorption capacities (approximately 100 microg/g) comparable to that of the activated carbon; they showed far superior SO2 and NOx sorption capacities. Preliminary cost estimates for the process utilizing these novel sorbents indicate potential for substantial lowering of control costs, as compared with other processes currently used or considered for control of Hg0, SO2, and NOx emissions from coal-fired boilers. The implications of these findings toward development of multipollutant control technologies and planned pilot and field evaluations of more promising multipollutant sorbents are summarily discussed. PMID- 11924859 TI - Effects of nitrogen and oxygen on biofilter performance. AB - Three laboratory-scale biofilters packed with inert material were used to study the nitrogen and oxygen requirements for biofiltration of methanol. Mixtures of methanol with inorganic nitrogen (NH3 or NO3) at nitrogen-to-carbon (N:C) ratios ranging from 0.015 to 0.4 were employed to reveal nitrogen effects on biofiltration. In the oxygen study, mixtures of air and oxygen at different oxygen contents were used. At low nitrogen levels, the removal rate increased with increasing N:C ratio for both NH3 and NO3. However, at high concentrations, NH3 had an inhibitory effect on biodegradation while the removal rate reached a plateau at high NO3 concentrations. Biofiltration with 63% oxygen in the inlet gas stream increased the maximum removal rate from 120 to 145 g/m3/hr after 3 days in comparison with biofiltration with air. However, a further increase in oxygen content up to 80% did not lead to a further improvement in biofilter performance, suggesting that both oxygen and biofilm thickness can be the relevant factors limiting biofilter performance and creating the plateau in removal rates at high loadings. PMID- 11924860 TI - Respirable particulate matter at an urban and nearby industrial location: concentrations and variability and synoptic weather conditions during high pollution episodes. AB - Continuous data of the concentration measurements of respirable suspended particulates (PM10, particles with aerodynamic diameter smaller than or equal to 10 pm) were analyzed. These measurements were carried out at an urban and nearby industrial location in northern Greece for the 5-year period 1996-2000. The time series concentration trend was examined, the seasonal and diurnal variations were identified, and the lognormality of the daily mean concentration data sets was tested. Over the 5-year data-gathering period, the days on which the U.S. Environmental Protection Agency (EPA) 24-hr PM10 standard was exceeded (episode days) were identified and their relation to prevailing synoptic-scale meteorological conditions was studied. The analysis led to useful information concerning the air quality levels, the contribution of the main pollution sources in this area, as well as some of the mechanisms that influence the PM10 concentrations. It also was proved that the measured PM10 concentrations are a result of a combination of processes including local anthropogenic sources, mesoscale transport, and resuspension. A complex system of sources and meteorological conditions modulate the heavy particulate pollution in the area of interest. PMID- 11924861 TI - Size distribution and diurnal and seasonal trends of ultrafine particles in source and receptor sites of the Los Angeles basin. AB - This paper presents results from a study conducted in two urban areas of southern California, Downey and Riverside, to examine the effect of different sources and formation mechanisms on the size distribution and temporal trends of ultrafine particles. Near-continuous data were collected for 5 months at each location. Our data clearly identified Downey as a source site, primarily affected by vehicular emissions from nearby freeways, and Riverside as a receptor site, where photochemical secondary reactions form a substantial fraction of particles, along with local vehicular emissions. In Downey, the diurnal trends of total particle number concentration and elemental carbon (EC) appear to be almost identical throughout the day and irrespective of season, thereby corroborating the role of primary emissions in the formation of these particles. This agreement between EC and particle number was not observed in Riverside during the warmer months of the year, while very similar trends to Downey were observed during the winter months in that area. Similarly, the size distribution of ultrafine particles in Downey was generally unimodal with a mode diameter of 30-40 nm and without significant monthly variations. The number-based particle size distributions obtained in Riverside were bimodal, with a significant increase in accumulation mode as the season progressed from winter to summer. During the warmer months, there was also an increase in sub-100-nm particles in the afternoon hours, between 2:00 p.m. and 4:00 p.m., that also increased with the temperature. The differences observed in the ultrafine particle distribution and temporal trends clearly demonstrated that mechanisms other than direct emissions play an important role in the formation of ultrafine particles in receptor sites of the Los Angeles Basin. PMID- 11924862 TI - Prediction of pressure drop in an orifice scrubber based on a Lagrangian approach. AB - A mathematical model has been developed to predict pressure drop in an orifice scrubber. This model is based on a Lagrangian approach for droplet movement and a particle-source-in-cell (PSI-CELL) model for calculating droplet concentration distribution. The k-epsilon turbulent model including body force due to the drag force between fluid and droplets was used to evaluate the fluid velocity distribution. The effect of orifice size on pressure drop and the correlations for mean droplet diameter have been studied. The results from the model have been compared with experimental data. This comparison shows excellent agreement between the calculated results and the experimental data. PMID- 11924863 TI - An atmospheric dispersion model for the environmental impact assessment of thermal power plants in Japan--a method for evaluating topographical effects. AB - An atmospheric dispersion model was developed for the environmental impact assessment of thermal power plants in Japan, and a method for evaluating topographical effects using this model was proposed. The atmospheric dispersion model consists of an airflow model with a turbulence closure model based on the algebraic Reynolds stress model and a Lagrangian particle dispersion model (LPDM). The evaluation of the maximum concentration of air pollutants such as SO2, NOx, and suspended particulate matter is usually considered of primary importance for environmental impact assessment. Three indices were therefore estimated by the atmospheric dispersion model: the ratios (alpha and beta, respectively) of the maximum concentration and the distance of the point of the maximum concentration from the source over topography to the respective values over a flat plane, and the relative concentration distribution [gamma(x)] along the ground surface projection of the plume axis normalized by the maximum concentration over a flat plane. The atmospheric dispersion model was applied to the topography around a power plant with a maximum elevation of more than 1,000 m. The values of alpha and beta evaluated by the atmospheric dispersion model varied between 1 and 3 and between 1 and 0.4, respectively, depending on the topographical features. These results and the calculated distributions of y(x) were highly similar to the results of the wind tunnel experiment. Therefore, when the slope of a hill or mountain is similar to the topography considered in this study, it is possible to evaluate topographical effects on exhaust gas dispersion with reasonable accuracy using the atmospheric dispersion model as well as wind tunnel experiments. PMID- 11924864 TI - On the relationship between acid rain and cloud type. AB - Spanish European Monitoring and Evaluation Programme (EMEP) stations were selected to relate acid rain episodes with the meteorological structure that caused the rainfall during a 5-year period. A principal component analysis (PCA) was used to determine the origin of major ions (SO4(2-), NO3-, Cl-, Ca2+, K+, Mg2+, and Na+) in the rainwater. In addition, the meteorological origin of the rain was identified. Previous works suggested a relationship between acid rain and storm convective clouds. However, statistical analyses of pH values show that only the short-lived convective phenomena may cause acid rain in Spain. In fact, rain generated by fronts and that related to long-lived convective systems is neutral or even slightly basic. Results suggest that the acid rain might be related to the meteorological time scale instead of to the cloud type. PMID- 11924865 TI - Modeling assessment of air emission flux of mercury from soils in terrestrial landscape components: model tests and sensitivities. AB - The abilities of a screening-level model to predict variations in elemental mercury (Hg0) air emissions from soils in terrestrial landscapes are examined by comparing simulation results to published observational data and by performing sensitivity analyses. Despite uncertainties and simplifications, the model results obtained offer some degree of confidence in the model's joint ability to relate readily available environmental parameters to airborne emissions of Hg predicted by coupling simple atmospheric and soil parameters with Hg cycling and transport algorithms. The model reasonably predicted the observational data in the considered data sets except for one site for which significant uncertainty was associated with model input data. Predictions are consistent with many trends observed in the field studies; better predictions were obtained for nonvegetated systems (relative errors between 0.4 and 9.7%) than for shaded-soil landscapes (relative errors between 2.3 and 27%). The model reflected field data showing that daily average emission rates of Hg0, formed by the reduction of Hg(II), are primarily controlled by changes in solar radiation, soil moisture, temperature, and, to a lesser extent, wind conditions. The model may have potential use in several preliminary studies to characterize trends of airborne Hg emitted from terrestrial sources to the atmosphere. PMID- 11924866 TI - Measurement and three-dimensional modeling of airflow and pollutant dispersion in an undersea traffic tunnel. AB - Airflow and pollutant dispersion in a cross-harbor traffic tunnel were experimentally and numerically studied. Concentrations of the gaseous pollutants CO, NOx, and total hydrocarbons (THC) at three axial locations in the tunnel, together with traffic flow rate, traffic speed, and types of vehicle were measured. Three-dimensional (3D) turbulent flow and dispersion of air pollutants in the tunnel were modeled and solved numerically using the finite volume method. Traffic emissions were modeled accordingly as banded line sources along the tunnel floor. The results reveal that cross-sectional concentrations are nonuniformly distributed and that concentrations rise with downstream distance. The piston effect of vehicles alone can provide 9-23% dilution of air pollutants in the tunnel, compounded to a 23-74% dilution effect according to the ventilation condition. PMID- 11924867 TI - Sleep deprivation in prion protein deficient mice sleep deprivation in prion protein deficient mice and control mice: genotype dependent regional rebound. AB - We have previously reported a larger and more prolonged increase of slow wave activity (SWA) in NREM sleep after sleep deprivation (SD) in prion protein deficient mice (PrP) compared to wild-type mice. Regional differences in the SWA increase were investigated by comparing the effect of 6 h SD on a frontal and occipital derivation in PrP deficient mice and wild-type mice. The larger increase of SWA after SD in PrP deficient mice was restricted to the occipital derivation. The difference appeared after the waking-NREM sleep transitions, making it unlikely that PrP is involved in the mechanisms enabling the transition to sleep. Our findings may reflect differences between the genotypes in the need for recovery in this particular brain region. PMID- 11924869 TI - Regulation of dynorphin gene expression by kappa-opioid agonist treatment. AB - The effects of K-opioid agonist treatment on prodynorphin mRNA expression in the rat brain were studied. Rats were treated with the selective kappa-opioid agonist U-69593 or vehicle for 5 days and prodynorphin mRNA was measured on day 8 (3 days after the last injection) or 22 (17 days after the last injection). On day 8 prodynorphin mRNA was increased in the hypothalamus and decreased in the striatum, frontal cortex, and hippocampus of rats treated with U-69593. On day 22, prodynorphin mRNA was increased in the hypothalamus, frontal cortex and striatum of U-69593 treated rats. These findings suggests that kappa-opioid receptor agonist treatment has long-term, continually changing effects on prodynorphin mRNA expression. PMID- 11924868 TI - Apoptotic genes expression in the lumbar dorsal horn in a model neuropathic pain in rat. AB - This study combines behavioural, molecular and morphological approaches to assess the occurrence of apoptosis in the rat spinal cord by 14-day sciatic nerve chronic constrictive injury (CCI). Thermal allodynia developed in the corresponding footpad 2-3 days after surgery, while morphological features, evaluated 14 days later, consisted in a decrease (23 +/- 7%) in laminae I-III cell number ipsilateral to CCI. Apoptosis occurrence was possibly suggested by the presence of some TUNEL-positive nuclei in this territory. The mRNA expression levels of the bcl-2 genes family was changed as follows: bax increased up to 40% in CCI vs the sham rats, while bcl-2 did not change; bcl-xS massively decreased (by 70% and 100%), while bcl-xL increased (by 40%) in CCI rats. Western blot analysis showed no change either on poly-ADP ribose polymerase (PARP) or p53 transcription factor in CCI and sham rats. These data suggest that in a chronic pain condition, where the acute phase has already resolved, specific apoptotic genes are still operative and possibly may serve as a critical change for cells surviving in the chronic pain state. PMID- 11924870 TI - Chronic paroxetine increases [3H]nociceptin binding in rat dorsal raphe nucleus. AB - Opioid-receptor-like I (ORLI) receptors, ORLI mRNA and nociceptin are particularly abundant in the limbic system and in central monoaminergic areas, brain regions involved in mood regulation and response to antidepressants. To analyse whether ORLI receptors adaptations occur during the first 3 weeks of an antidepressant treatment, we administered paroxetine to rats (10 mg/kg, i.p., once a day) for 4, 7,14 or 21 days. A significant increase (22-50%) in [3H]nociceptin binding sites density appeared specifically in the dorsal raphe nucleus after 4, 7 or 21 days of treatment, whereas no change was observed at any time in any other brain regions studied. These data constitute the first evidence of a modulation of ORLI receptors by an antidepressant treatment. PMID- 11924872 TI - The acute effects of exposure to the electromagnetic field emitted by mobile phones on human attention. AB - The aim of our study was to investigate the effects of acute mobile phone exposure on a range of tasks which tapped capacity and processing speed within the attentional system. Thirty-eight healthy volunteers were randomly assigned to either an experimental group which was exposed to a connected mobile phone or a control group in which the mobile phone was switched off. Subjects remained blind to mobile phone status throughout duration of study. The experimental group were exposed to an electromagnetic field emitted by a 900 MHz mobile phone for 30 min. Cognitive performance was assessed at three points (prior to mobile phone exposure, at 15 and 30 min post-exposure) using six cognitive neuropsychological tests (digit span and spatial span forwards and backwards, serial subtraction and verbal fluency). Significant differences between the two groups were evident after 5 min on two tests of attentional capacity (digit span forwards and spatial span backwards) and one of processing speed (serial subtraction). In all three instances, performance was facilitated following mobile phone exposure. No deficits were evident. These findings are discussed in terms of possible functional and neuroanatomical bases. PMID- 11924871 TI - Tissue plasminogen activator is required for striatal post-ischemic synaptic potentiation. AB - Recent experimental observations indicate that tPA plays a key role in the development of neuronal damage that follows cerebral ischemia and excitotoxicity. In an attempt to clarify how tPA favors ischemia-induced neuronal damage, we performed in vitro electrophysiological experiments in striatal slices by using mice selectively lacking this serine protease.We found that tPA ablation did not affect the membrane depolarization of striatal neurons exposed to combined oxygen and glucose deprivation but fully prevented the induction of NMDA-dependent post ischemic long-term synaptic potentiation. The absence of striatal post-ischemic pote ntiat ion observed in tPA-lacking mice may account for the significant neuroprotection observed in these animals after the occlusion of middle cerebral artery. PMID- 11924874 TI - Expression of protein kinase C in song control nuclei of deafened adult male Bengalese finches. AB - Auditory feedback is necessary to maintain singing in the adult male Bengalese finch (songbirds/oscines). Their song patterns are altered within a month following cochlear removal-induced deafness. Stabilization of song patterns occurs thereafter. To clarify what kind of changes appear in the brain of deafened birds, we examined immunohistochemically the expression of protein kinase C (PKC), considered a molecular marker for synaptic plasticity, in the robust nucleus of the archistriatum (RA), one of the song control nuclei in the forebrain of finches. Two weeks after cochlear removal, immunoreactive fibers and terminals in the RA transiently increased, and thereafter tended to decrease gradually. Moreover, the degree of song alteration and stability paralleled these changes in the RA. The immunoreactivity of the RA remained unchanged in intact birds. These results indicate that surgical deafening increases the expression of PKC in the RA. These changes in the RA are related to the alteration of song patterns in the deafened adult Bengalese finch. PMID- 11924873 TI - MRI and neuropsychological improvement in Huntington disease following ethyl-EPA treatment. AB - A 6-month randomized, placebo-controlled pilot study of the ethyl-ester of eicosapentaenoic acid (ethyl-EPA) was carried out in seven in-patients with advanced (stage III) Huntington's disease (three on ethyl-EPA, four on placebo; no significant difference in age or sex between the groups). After 6 months all the patients treated with ethyl-EPA improved on the orofacial component of the Unified Huntington's Disease Rating Scale while all the patients on placebo deteriorated on this scale (p < 0.03). Following subvoxel registration of follow up 3D MRI brain scans with baseline scans, subtraction images showed that while the placebo was associated with progressive cerebral atrophy, the ethyl-EPA was associated with a reverse process. We conclude that treatment with ethyl-EPA is associated with beneficial motor and MRI changes. PMID- 11924875 TI - Implicit learning of emotional information under anesthesia. AB - Surgical stress activates the amygdala and secretion of norepinephrine, both involved in memory and unconscious processing of emotionally negative information. This study examined surgically induced facilitation of implicit learning of emotionally negative information. Thirty patients undergoing laparoscopic cholecystectomy under general anesthesia were tested. Between 2 and 4 h after surgery, patients provided word-associates to cues previously presented (old) or not previously presented during anesthesia (new). Half the cues were emotionally negative and half neutral. Patients took less time to provide correct associates to old emotionally negative cues than to new emotionally negative cues (p < 0.05). Spectral edge frequency (SEF) of cerebral activity during surgery converged with this finding. Implicit learning during general anesthesia may be stronger for emotionally negative information and is detected by SE. PMID- 11924876 TI - Neuronal activity in primate striatum and pallidum related to bimanual motor actions. AB - To assess whether striatal and pallidal neurones may contribute to bimanual co ordination, two macaque monkeys were trained to perform a delayed conditional sequence of co-ordinated pull and grasp movements, executed either bimanually or unimanually. Most of the 58 task-related neurones, recorded from the caudate nucleus, putamen, external and internal divisions of the globus pallidus, exhibited an activity related to the execution of the movements. Only a quarter of neurones displayed preparatory activity. The majority of units exhibited a significant modulation of activity in unimanual trials irrespective of the hand used to perform the task. In bimanual trials, one-third of units exhibited discharge patterns reflecting a bimanual synergy, suggesting a possible role for basal ganglia in inter-limb co-operation. PMID- 11924877 TI - Getting ready to remember: the neural correlates of task set during recognition memory. AB - Event-related potentials (ERPs) were employed to investigate the neural correlates of episodic and semantic task sets. ERPs elicited by cues signalling an upcoming recognition memory test trial showed a sustained positivity relative to those signalling an upcoming semantic test trial, lasting from 500 ms post-cue until the arrival of the test item. However, this effect was present only on the second successive trial on which subjects performed the recognition task. Thus, when episodic vs semantic tasks vary trial-by-trial, the establishment of a recognition memory task-set is not achieved within a single trial. The findings are discussed in relation to the notion of episodic retrieval mode. PMID- 11924879 TI - Vestibulo-ocular, optokinetic and postural function in diabetes mellitus. AB - We compared vestibulo-ocular reflex, optokinetic reflex and postural function in subjects with insulin-dependent diabetes mellitus (IDDM) and non-insulin dependent diabetes mellitus (NIDDM), as well as non-diabetic controls. Both IDDM and NIDDM subjects exhibited significant deficits in gaze-holding in darkness (p < 0.05), small changes in vestibulo-ocular reflex (VOR) phase re velocity (p < 0.005) without a change in VOR gain, and a decrease in optokinetic reflex (OKR) slow phase velocity (p < 0.001). In addition, a smaller decrease was found in OKR quick phase amplitude (p < 0.02); postural sway was increased in both diabetic groups (p < 0.05), although this was not specific to the conditions of the Clinical Test of Sensory Interaction and Balance (CTSIB) that test vestibular contributions to postural stability. No differences were found in optokinetic afternystagmus or latency to circularvection. These results suggest that both IDDM and NIDDM are associated with deficits in gaze-holding, VOR and OKR function. PMID- 11924878 TI - Positive and negative emotional verbal stimuli elicit activity in the left amygdala. AB - The human amygdala's involvement in negative emotion is well established, but relatively little is known regarding its role in positive emotion. Here we examined the neural response to emotionally positive, negative, and neutral words using fMRI. Relative to neutral words, positive and negative emotional words elicited greater activity in the left amygdala. Positive but not negative words elicited activity in dorsal and ventral striatal regions which have been linked in previous neuroimaging studies to reward and positive affect, including caudate, putamen, globus pallidus, and accumbens. These findings provide the first direct evidence that the amygdala is involved in emotional reactions elicited by both positive and negative emotional words, and further indicate that positive words additionally activate brain regions related to reward. PMID- 11924880 TI - Scopolamine impairs eyeblink conditioning in cerebellar LTD-deficient mice. AB - We investigated the effect of the muscarinic acetylcholine receptor (mAChR) antagonist scopolamine on eyeblink conditioning in glutamate receptor subunit 62 null mice, which have severe impairments in cerebellar long-term depression (LTD). Mice were injected intraperitoneally with scopolamine (0.5 mg/kg) or saline, and conditioned using a delay paradigm with tone and periorbital shock but with no overlap between them. The saline-injected mutant mice learned this paradigm normally, as predicted from our previous study. When scopolamine was injected, learning was impaired more severely in the mutant mice than in the wild type mice. Basic sensory and motor performances were not affected. These results suggest that eyeblink conditioning in cerebellar LTD deficient mice depends largely on neural functions susceptible to blocking of mAChRs. PMID- 11924881 TI - Does ethanol activate G-protein coupled inwardly rectifying K+ channels? AB - G-protein coupled inwardly rectifying K+ (GIRK) channels have been reported to be targets of ethanol actions. We investigated if ethanol affects native GIRK channels in rat brain tissues at clinically relevant concentrations using brain slices containing the ventrolateral periaqueductal gray (PAG), an area related to pain regulation. Ethanol did not affect the membrane current elicited by hyperpolarization ramps at concentrations up to 150 mM. However, at 200-300 mM, which is above the lethal level, it activated a barium-sensitive GIRK current in 30-57% of neurons. In neurons unresponsive to ethanol, baclofen, the mu-opioid or nociceptin successfully activated GIRK channels. It is suggested that GIRK channels of the ventrolateral PAG are unlikely to be targets of the analgesic action of ethanol. PMID- 11924882 TI - A comparison of extracellular levels of phenytoin in amygdala and hippocampus of kindled and non-kindled rats. AB - Temporal lobe epilepsy is often refractory to antiepileptic drugs (AEDs). Accordingly, amygdala-kindled rats, a widely used model of temporal lobe epilepsy, have previously been found to be less responsive to AED treatment than non-kindled rats. In view of recent findings of over-expression of multidrug transporters in the blood-brain barrier of patients with pharmacoresistant epilepsy, one explanation for the finding of difficult-to-treat seizures in kindled rats would be a reduced penetration of AEDs into epileptogenic brain tissue. For evaluation of this possibility, we used brain microdialysis in order to compare extracellular levels of the AED phenytoin in amygdala and hippocampus of conscious, unrestrained amygdala-kindled and non-kindled rats. Consistent with the lower anticonvulsant efficacy of phenytoin in kindled rats, average phenytoin levels in dialysates of kindled rats were lower (up to 30%) than in non-kindled controls, but the differences were not statistically significant. This indicates that either the relatively large interindividual variation in dialysate levels of phenytoin masks functionally significant differences in individual kindled rats or that alterations in brain drug penetration are not involved in the lowered response of kindled rats to AEDs. PMID- 11924883 TI - A dopamine D3 receptor partial agonist blocks the expression of conditioned activity. AB - The partial dopamine D3 receptor agonist BP 897 attenuates cocaine seeking suggesting that BP 897 will attenuate conditioned activity to environmental stimuli paired with amphetamine. During conditioning, amphetamine (2.0 mg/kg) stimulated activity and co-treatment with BP 897 (1.0 mg/kg) had no effect. In the saline test, groups conditioned with amphetamine or amphetamine plus BP 897 showed conditioned activity. Treatment with BP 897 in the test following conditioning with saline produced no significant effect but following conditioning with amphetamine BP 897 blocked conditioned activity. Results extend previous findings that BP 897 attenuates responding for cocaine-paired stimuli to amphetamine-paired stimuli in a different paradigm and support the potential of BP 897 as a therapeutic agent for the prevention of drug seeking. PMID- 11924884 TI - Memory-based comparison process not attenuated by haloperidol: a combined MEG and EEG study. AB - Auditory P50 and N100 responses reflect preattentive processing, whereas subsequent mismatch negativity (MMN) response indexes memory-based comparison process. Divergent ERP responses have been found in schizophrenia and in Parkinson's disease (PD), which have abnormalities in cerebral dopamine activity. We used simultaneously magnetoencephalography and electroencephalography to investigate, whether a single dose of haloperidol, a dopamine D2-receptor antagonist, modulates preattentive auditory processing using a randomized, double blind, placebo-controlled crossover design. Our results showed that haloperidol did not alter MMN to frequency and duration changes, whereas the magnetic MMN to frequency change was significantly accelerated. The amplitude and latency changes of the electric and magnetic P50 and N100 were insignificant. Our results indicate that memory-based sound comparison and preceding cortical processing underlying stimulus detection are not attenuated by haloperidol, whereas haloperidol appears to accelerate preattentive sound comparison. PMID- 11924885 TI - Proton MR spectroscopic study at 3 Tesla on glutamate/glutamine in Alzheimer's disease. AB - To investigate metabolic changes in Alzheimer's disease (AD), we performed proton MR spectroscopy at 3T Spectra were acquired from the gray matter of the posterior cingulate gyrus and the precuneus, and from the parietooccipital white matter in nine AD patients and 12 controls. In patients, the N-acetyl group (NA)/creatine + phosphocreatine (Cr) ratios were decreased in both regions, and a decrease in the glutamate + glutamine (Glx)/Cr ratio and a correlation between the NA/Cr and Glx/Cr ratios were detected in the gray matter, but not in the white matter. These results suggest that NA and Glx metabolism are simultaneously affected in AD, however, metabolic changes in Glx are more profound in the gray matter than in the white matter. PMID- 11924886 TI - Tonic gravity changes alter gene expression in the efferent vestibular nucleus. AB - To assess effects of tonic gravity changes on efferent vestibular neurons, immediate early gene (IEG) protein expression was compared in the efferent vestibular nucleus (EVe) of adult rats exposed to microgravity during the NASA Neurolab Mission (STS-90), and matching control animals on the ground. Rats sacrificed 1 day and 12 days after launch (experiencing approximately 0g) showed significantly fewer IEG protein-expressing cells than corresponding ground control animals (at 1 g). Normal IEG protein-expressing cell numbers reappeared within I day after flight animals returned to earth, and were maintained at 14 days after landing. EVe neurons appear to play a role in gravity-induced modifications of vestibular hair cell synapses; a contribution to gravity-induced plasticity of the vestibulo-ocular reflex (VOR) is also suggested. PMID- 11924887 TI - PAF antagonist treatment reduces pro-inflammatory cytokine mRNA after spinal cord injury. AB - Platelet-activating factor (PAF) is a pro-inflammatory molecule which contributes to secondary damage after spinal cord injury (SCI). To test if PAF contributes to cytokine induction following SCI, female Long-Evans rats were pretreated with the PAF antagonist WEB 2170 prior to receiving a contusion injury at spinal cord level T10 using the NYU impactor. RNase protection assay (RPA) analysis revealed that IL-1alpha mRNA peaked at I h post-injury while IL-1beta and IL-6 mRNA levels were higher and peaked at 6 h.TNF-alpha mRNA was almost undetectable. All mRNA levels approached baseline by 24 h. Treatment with WEB 2170 (1 mg/kg, i.p.) 15 min prior to injury significantly decreased mRNA levels for all three cytokines at 6 h post-injury, but not at I h post-injury. These results demonstrate a role for PAF in proinflammatory cytokine induction after SCI. PMID- 11924888 TI - N-methyl-norsalsolinol, a putative dopaminergic neurotoxin, passes through the blood-brain barrier in vivo. AB - In earlier studies the dihydroxylated tetrahydroisoquinoline derivatives salsolinol and 2(N)-methyl-norsalsolinol (NMNorsal), a 2(N)-analogue of salsolinol, were identified as putative endogenous neurotoxins in patients with Parkinson's disease. Since a prominent blood-brain barrier (BBB) was described to exist for salsolinol, in the present study microdialysis experiments were performed to investigate the penetration of NMNorsal through the BBB into the caudate nucleus of the rat brain. After i.p. administration of NMNorsal (20 mg/kg), it could be detected in the dialysate of the caudate nucleus with a mean maximum after 40 min. There was no alteration in extracellular dopamine or 3,4 dihydroxyphenylacetic acid levels. Addition of the monoamine oxidase inhibitor pargyline (10 microM) to the perfusate did not modify NMNorsal levels in the caudate nucleus. To corroborate the microdialysis results, homogenates of the contralateral caudate nucleus were prepared and NMNorsal could also be detected. These findings indicate that NMNorsal is indeed able to pass through the blood brain barrier of the rat brain. PMID- 11924889 TI - A randomised, placebo-controlled, double blind study of treatment of Huntington's disease with unsaturated fatty acids. AB - Huntington's Disease (HD) is a serious dominantly inherited neurodegenerative disorder for which there are no current treatments. Open label and animal studies have suggested that highly unsaturated fatty acids (HUFAs) may be beneficial. Seventeen patients with HD were entered into a randomised, placebo-controlled, double blind trial of HUFA therapy. Patients were assessed on the Rockland Simpson Dyskinesia Rating Scale (RSDRS) and the Unified Huntington's Disease Rating Scale (UHDRS). On the RSDRS and the UHDRs motor scale patients on HUFA treatment improved while those on placebo deteriorated, with a significant difference between the two groups on the RSDRS. A similar trend was noted on the UHDRS functional performance scales. Little change was seen on the neuropsychology scales. There were no treatment-related adverse events. This is the first time that a significant improvement has been noted in a randomised trial in HD. The results are consistent with open label observations; a second placebo-controlled study in end-stage patients, and a study in a transgenic mouse model of HD. PMID- 11924890 TI - TGF-beta1-conditioned glial cell-derived dendritic cells inhibit expansion of MBP reactive T cells in vitro. AB - Resident microglial cells contribute to activation and expansion of T cells under inflammatory conditions within the CNS. However, there is no evidence how interactions between microglia and T cells affect CNS inflammation. We evaluated the effect of glial cell-derived dendritic cells (GC-DC) in expanding and eliminating myelin basic protein (MBP)-reactive T cells. GC-DC untreated with TGF beta1 (GC-DC0) primed antigen specific T cell proliferation, whereas GC-DC treated with TGF-1 (GC-DCbeta) effectively inhibited expansion of T cells via inducing IFN-y-expressing CD8+ T cells. Augmented IFN-gamma and/orTNF-alpha might also affect the elimination of MBP-reactive T cells. These results indicate that TGF-beta1-mediated functional skewing of GC-DC plays a critical role for the elimination of MBP-reactive T cells. PMID- 11924891 TI - Large-scale cortical displacement of a human retinotopic map. AB - The aim of the current study was to determine the retinotopic organization of a patient with congenital cortical dysgenesis and normal visual function. Using functional magnetic resonance imaging (fMRI), detailed retinotopic maps corresponding to the four visual field quadrants were projected onto cortical surfaces. Similar to control subjects, the upper right visual field mapped onto ventral left hemisphere and was retinotopically organized. The lower right visual field's cortical representation was also retinotopically organized, yet was displaced many centimeters anteriomedially. Moreover, the entire left visual field was represented in non-retinotopically organized islands in both hemispheres. These results indicate retinotopic maps can shift in both location and topography illustrating cortical reorganization presumably due to either cortical dysgenesis or functional displacement. NeuroReport PMID- 11924892 TI - Maturation of the auditory event-related potentials during the first year of life. AB - This study examined the maturation of cortical auditory event-related potentials (ERPs) from birth until 12 months of age. In the 15 infants studied, all ERP peaks observable at 12 months of age, the P150, N250, P350, and N450 were identifiable already at birth, As in previous studies, the amplitudes of the ERP peaks increased and latencies shortened with increasing age. In addition, the time courses of the amplitude growth of these peaks differed from each other. It was concluded, that the generators of all the infantile ERP peaks are functional already at birth, and that the maturational changes in the waveform morphology can mostly be accounted for by the changing relative strengths of the different generators. PMID- 11924893 TI - Striatal tachykinin and enkephalin mRNAs are normalized by serotonin2 and NMDA manipulation following dopamine depletion. AB - We have examined the effects of serotonin2 (5-HT2) stimulation and NMDA antagonism on preprotachykinin (PPT) and preproenkephalin (PPE) gene regulation in the dopamine (DA) depleted striatum. Following DA lesions, PPT mRNA expression was reduced (dorsomedial (DM) 44 +/- 9%, dorsolateral (DL) 40 +/- 4%), whereas PPE message levels were elevated (DM 207+/-28%, DL 198+/-25%). Within this state of dysregulated gene activity, DOI (5-HT2 agonist) increased PPT message levels (174 +/- 5%, DM; 153 +/- 13%, DL) without affecting PPE gene expression. In addition, MK-801 (NMDA antagonist) decreased PPE message levels (DM 59 +/- 10%, DL 52 +/- 7%) without significantly altering PPT mRNA expression. Combined application of DOI and MK-801 resulted in normalization of both PPTand PPE message. Statistical analysis revealed no drug interactions in this paradigm suggesting independent mechanisms for 5-HT2 and NMDA receptors in controlling neuropeptide production following DA depletion. PMID- 11924894 TI - African trypanosomes activate human fetal brain cells to proliferation and IFN gamma production. AB - We addressed the host-parasite interplay and the immunopathogenetic events occurring in the central nervous system (CNS) during human African trypanosomiasis. Human first trimester forebrain cells were stimulated with a trypanosome lymphocyte-triggering factor (TLTF) and studied for their immune response as exemplified by cell proliferation and IFN-gamma production. TLTF induced proliferation of human first trimester forebrain cells and IFN-gamma production at the mRNA and protein levels. Astrocytes are the major producers of IFN-gamma in response toTLTE These data illustrated for the first time a direct effect of a parasite factor on human brain cells. TargetingTLTF during the course of the disease may be considered in preventing the deadly neurological complications of human African trypanosomiasis. NeuroReport PMID- 11924895 TI - SURI and Kir6.1 subunits of K(ATP)-channels are co-localized in retinal glial (Muller) cells. AB - ATP-sensitive potassium channels (K(ATP)), unlike other inwardly rectifying potassium (Kir) channels, require two structurally diverse subunits to form functional channels: one member of the Kir6 channel family (Kir6.1 or Kir6.2), and one sulfonylurea receptor (SUR) of the ATP-binding cassette superfamily (SURI, SUR2A or SUR2B). We have previously shown that two pore-forming subunits of K(ATP)-channels are differently distributed in frog retina. Kir6.1 is localized in Miller (glial) cells, whereas Kir6.2 is found in neurons. Using immunocytochemistry, the present study reveals that in adult frog retina, SURI is restricted to Muller (glial) cells whereas SUR2A and SUR2B are found in neurons. These data suggest that functional K(ATP) channels in Muller cells may be formed by Kir6.1/SURI, and in neurons by Kir6.2/SUR2A and/or Kir6.2/SUR2B. PMID- 11924896 TI - Optokinetic stimulation and the egocentred midsagittal plane: an fMRI study. AB - Animal studies and observations of neglect patients suggest that the posterior parietal cortex is part of a system that codes ego-centred space. Few studies show the existence of areas involved in the representation of egocentred space in healthy humans. We investigated with fMRI and a conjunction protocol, the overlap of activity between optokinetic stimulation and a task of midline computation. Results showed that the right posterior parietal and frontal cortices were involved in both tasks (p < 0.0001). The evidence presented in this study provides the neuroanatomical substrate involved in the recovery of neglect during vestibular stimulation. This last point is of clinical interest for the rehabilitation of hemineglect. PMID- 11924897 TI - Neuroanatomical substrata of amusement and sadness: a PET activation study using film stimuli. AB - The aim of the present study was to explore the neuroanatomical substrata of film induced amusement and sadness, aiming at maximal reliability by using strict statistics. Film-induced changes in regional cerebral blood flow (rCBF) were measured using positron emission tomography and [15O]water. It appeared that many brain regions were involved similarly and symmetrically in both emotions. Activation of the occipitotemporal and anterior temporal cortex and cerebellum during both emotions agreed well with the earlier findings. Contrary to some earlier studies, the posterior cingulate gyrus and some frontal areas were deactivated in both emotions. The strict explorative analysis did not reveal activations in the limbic or subcorticai structures seen in some earlier studies. However, an additional analysis restricted to the subcortical and limbic system structures revealed bilateral activation of the amygdala in both target emotions. The outcomes of imaginal studies of emotions seem to be largely dependent on the conservativeness of the statistical analysis and very likely also on the method of emotion induction. PMID- 11924898 TI - Neuroprotection in the rat Parkinson model by intrastriatal GDNF gene transfer using a lentiviral vector. AB - We used a recombinant lentiviral vector (rLV) for gene delivery of GDNF to the striatum, and assessed its neuroprotective effects in the intrastriatal 6 hydroxydopamine (6-OHDA) lesion model. The level of GDNF expression obtained with the rLV-GDNF vector was dose-related and ranged between 0.9-9.3 ng/mg tissue in the transduced striatum, as determined by ELISA, and 0.2-3.0 ng/mg tissue were detected in the ipsilateral substantia nigra (SN), due to anterograde transport of the GDNF protein. GDNF expression was apparent at 4 days and maintained for > 8 months after injection. Striatal delivery of rLV-GDNF efficiently protected the nigral dopamine (DA) neurons and their projection, against the 6-OHDA lesion (65 77% of intact side). Sprouting of the lesioned axons was observed along the nigrostriatal pathway, precisely corresponding to the areas containing anterogradely transported GDNF. PMID- 11924899 TI - Cerebral blood flow and cognitive responses to rivastigmine treatment in Alzheimer's disease. AB - Twenty seven patients with mild AD were enrolled in a prospective open label controlled study of rivastigmine. Assessments included a range of neuropsychiatric and behavioural measures and rCBF using HMPAO SPECTat baseline, three and six months. Significant enhancement of frontal, parietal and temporal brain blood flow with related psychometric improvement was observed in twelve of the treated patients. A pattern of reduced rCBF and cognitive performance was observed in four unresponsive and eleven untreated patients. The results suggest that alterations in the clinical and cognitive status of patients receiving a cholinesterase inhibitor are paralleled by changes in rCBF. Longitudinal assessment with repeated imaging offers a method of better understanding the effects of cholinesterase inhibition on the AD brain. PMID- 11924900 TI - Effect of melatonin on rat spinal cord nociceptive transmission. AB - Melatonin has been shown to exert potent antinociception but the sites and mechanisms of action underlying this effect have not yet been clarified. The effect of melatonin on spinal cord nociceptive transmission was studied in rats by assessing wid-up activity in a C-fiber reflex responses paradigm evoked by repetitive (0.6 Hz) electric stimulation. Intraperitoneal administration of 1.25, 2.5, 5.0 and 10.0 mg/kg melatonin induced a dose-dependent inhibition of spinal wind-up activity, the higher dose of the drug used being able to depress completely the C reflex gain. Results indicate that melatonin markedly depresses spinal wind-up in rats, probably through hyperpolarization of dorsal horn neurons consecutive to melatonin binding to membrane receptors, and/or via intracellular interference with a NMDA receptor-dependent nitric oxide generating pathway. PMID- 11924901 TI - MIG--differential gene expression in mouse brain endothelial cells. AB - Different diseases of the CNS are associated with blood-brain barrier (BBB) damage and mononuclear cell infiltration. In order to study genes that may play a role in endothelial cell regulation in inflammatory CNS diseases, we performed differential gene expression (DGE) analysis using a mouse brain endothelial cell line. We found that interferon-gamma (IFNgamma)-induced monokine (MIG), a chemokine that plays a role in T lymphocyte and monocyte chemoattraction, is highly expressed in the presence of inflammatory cytokines. We show that MIG, produced by brain endothelial cells in vitro, is biologically active in attractingT lymphocytes and that it is possible to interfere with this mechanism of action using anti-MIG antibodies. We suggest that blocking MIG may be beneficial in CNS inflammation. We detected constitutive expression of the MIG receptor, CXCR3, on the surface of the endothelial cells and therefore hypothesize that it plays a role in maintaining the cytokine gradient at the region of CNS inflammation. PMID- 11924902 TI - Central nociceptive role of prostacyclin (IP) receptor induced by peripheral inflammation. AB - Prostacyclin (PGI2) is well known to play crucial roles in induction of edema and pain behavior in the periphery. In the present study, we investigated the central role of PGI2 in inflammatory pain. Intraplantar injection of carrageenan markedly induced the expression of prostacyclin receptor (IP receptor) mRNA with the maximum at 6 h, coincidently induction of the inducible form of cyclooxygenase (COX-2), although IP receptor mRNA was weakly expressed in the spinal cord of naive mice. Intrathecal administration of the IP agonist cicaprost induced mechanical hyperalgesia 6 h after carrageenan injection. These results suggest that PGI2 is involved in pain transmission at the spinal cord following expression of IP receptor mRNA induced by peripheral inflammation. PMID- 11924903 TI - Transcallosal conduction time measured by visual hemifield stimulation with face images. AB - Transcallosal conduction time (TCT) in human visual system was estimated from the hemispheric difference of the peak latencies of face-evoked magnetic field (N170m) by left/right hemifield stimulation using 14 8 channel whole-head magnetometers. The estimated TCTwas 18.0 + 6.2 ms (mean +/- s.d.). Although N 170m amplitude was larger in the right hemisphere of seven of nine subjects, no hemispheric asymmetry was observed in the TCT estimate. Transcallosal visual information was likely to be transferred in the higher order (non-retinotopic) visual cortices. Hemifield stimulation with face images is a useful non-invasive method to estimate the TCT in human visual system. PMID- 11924904 TI - New concepts in neonatal seizures. AB - The immature brain is more prone to seizures than the older brain as a result of an imbalance between excitatory and inhibitory input. The depolarizing, rather than hyperpolarizing effect of GABA(A) during the first week of life in the rodent, and the delay in postsynaptic GABA(B) inhibition coupled with the over expression of glutamatergic synapses contribute to this increased propensity toward seizures. It is now clear that seizures can be injurious to the immature brain, although the pattern of seizure-induced injury is age-related. While the immature brain is resistant to acute seizure-induced cell loss, there are functional abnormalities following seizures with impairment of visual-spatial memory and reduced seizure threshold. Neonatal seizures are also associated with a number of activity-dependent changes in brain development including altered synaptogenesis and reduction in neurogenesis. These results argue that neonatal seizures should no longer be considered as benign events. PMID- 11924905 TI - Mammalian chemosensory receptors. AB - Chemosensory receptors are critical for the survival of many mammalian species, and their genes can comprise up to 1% of mammalian genomes. Odorant, taste, and vomeronasal receptors are being discovered and functionally characterized at a rapid pace which has been further accelerated by the availability of the human genome sequence. Five multigene families, consisting of >1,000 genes in the mouse, have been proposed to encode functional chemoreceptors. Although all of the chemoreceptor gene families encode G-protein coupled receptors, they are largely unrelated and uniquely specialized for the processing of different chemosensory modalities. Using members of the families as molecular probes, great insights are being gained into the different organizational strategies used by these sensory systems to encode information in both the periphery and the brain. PMID- 11924906 TI - Death receptor ligands in tumors. AB - Activation of apoptosis via death receptors is a tightly regulated event, and the death pathway itself is open to interference on the part of soluble or membrane bound decoy receptors. The aggregation state of the death-inducing ligand is a crucial factor, particularly when these molecules are used as recombinant drugs against tumors. Whether tumors are sensitive to such ligands is determined by both the net abundance of death receptors versus decoy receptors and the balance between intracellular apoptotic and antiapoptotic mechanisms. This means that in vivo elimination of tumor cells by effector arms such as T lymphocytes, natural killer cells, macrophages, and dendritic cells is dependent on both the function of activated lymphoid cells and the genetic properties of tumor cells. Death receptor ligands, however, may be a double-edged sword. When expressed on cytotoxic T lymphocytes, natural killer cells, monocytes, and dendritic cells, they induce the apoptosis of many tumor cells, whereas their expression on tumor cells induces the apoptosis of killer cells. The in vivo result is influenced by the number of infiltrating cells, their state of activation, the cytokine repertoire in the tumor microenvironment, and the ability of the tumor to produce soluble factors inhibiting their cytolytic functions. PMID- 11924907 TI - Promoter methylation controls the expression of MAGE2, 3 and 4 genes in human cutaneous melanoma. AB - Cancer-testis antigens expressed by different-histotype transformed cells are suitable targets for tumor immunotherapy. However, their heterogeneous expression in neoplastic lesions limits the eligibility of patients for cancer-testis antigen-directed vaccination, and low levels of cancer-testis antigens' expression may impair immune recognition of malignant cells. Because of the primary clinical relevance of cancer-testis antigens' expression in neoplastic tissues, 68 unrelated or sequential metastatic lesions from 56 patients were used to characterize the molecular mechanisms regulating the presence and levels of expression of different cancer-testis antigens of the MAGE family (i.e., MAGE2, 3 and 4) in cutaneous melanoma. Polymerase chain reaction-based methylation analyses showed that methylation status of specific cytosine-guanine dinucleotides in the promoters of investigated cancer-testis antigens correlated with their heterogeneous expression within unrelated metastatic melanoma lesions, and with their homogeneous expression among sequential metastases from three patients with melanoma. Unlike methylated promoters, unmethylated promoters of MAGE2, 3 and 4 genes drove the expression of reporter gene-enhanced green fluorescent protein after transient transfection of cancer-testis antigen positive Mel 142 melanoma cells. Furthermore, de novo expression of MAGE3 gene induced by the treatment of Mel 195 melanoma cells with the DNA hypomethylating agent 5-aza-2'-deoxycytidine was associated with a 6%-12% demethylation of selected cytosine-guanine dinucleotides in its promoter. Finally, 5-aza-2' deoxycytidine induced a 16-fold increase of MAGE3 expression in Mel 313 melanoma cells expressing constitutively low levels of the antigen, but did not affect that of Mel 275 melanoma cells expressing high baseline levels of MAGE3. Overall, these findings identify promoter methylation as a shared mechanism directly regulating the expression of therapeutic cancer-testis antigens in metastatic melanomas, and foresee the clinical use of 5-aza-2'-deoxycytidine to design new chemoimmunotherapeutic strategies in patients with melanoma. PMID- 11924908 TI - Antitumor effects of Flt3 ligand in transplanted murine tumor models. AB - Administration of Flt3 ligand (FL) to mice causes dendritic and natural killer cells to increase but certain solid tumors to regress. Depending on the particular tumor model used, T cells and natural killer cells have been implicated in the protective immune response induced by FL. The current study examined the effects of FL administration on tumor establishment and progression in metastatic and primary tumor models to correlate anatomic location with immunotherapeutic efficacy. FL mediated significant (p < or = 0.05) therapeutic activity against pulmonary metastases of the murine MC-38 colon adenocarcinoma, particularly when cytokine administration was initiated before tumor inoculation. However, progressive intraabdominal tumors sometimes were observed even in the relative absence of pulmonary metastases. Significant, although less dramatic, antimetastatic effects were observed with MCA-205 and MCA-102 sarcomas and D5 (B16BL6) melanoma. In contrast, FL was ineffective against subcutaneous MC-38 tumors or against several intracranial tumors. This suggests that besides the administration dose, the efficacy of this cytokine depends on the tumor type and possibly the location of the inoculated tumor. Antitumor activities of FL were abolished by whole-body irradiation (500 cGy) and partially abolished by systemic depletion of CD8, CD4, or natural killer cells. The results indicate that optimization of FL immunotherapy of tumors will require a firmer understanding of the relative contributions of tumor burden, location, immune system requirements, and other factors. PMID- 11924909 TI - Potential to target dysregulated interleukin-2 receptor expression in canine lymphoid and hematopoietic malignancies as a model for human cancer. AB - Lymphohematopoietic malignancies are common spontaneous diseases of dogs whose clinical presentation and biologic behavior closely resemble their human counterparts. The goal of this study was to define the potential to use canine lymphoma and leukemia as suitable models to refine therapeutic approaches targeting the interleukin-2 receptor (IL-2R). The authors evaluated the patterns of IL-2R expression in 13 dogs with multicentric non-Hodgkin's lymphoma (NHL) and in six dogs with leukemia (acute lymphocytic leukemia, n = 3; chronic lymphocytic leukemia in blast crisis, n = 1; acute monoblastic leukemia, n = 2). The authors first cloned and sequenced the complete coding domains of the wild-type canine IL 2R alpha-chain gene. They next used qualitative reverse transcription polymerase chain reaction (RT-PCR) analysis to examine IL-2R alpha, beta, and gamma(c) subunit expression in the tumors. Messenger RNA (mRNA) for the interleukin-2 receptor alpha, beta, and gammac subunits that comprise the high-affinity receptor was present in samples from all dogs with NHL. Expression of functional surface IL-2R also was observed flow cytometrically in NHL cells from all four dogs tested. Leukemic cells from one dog with B cell acute lymphocytic leukemia and two dogs with acute monoblastic leukemia expressed mRNA for all three subunits, whereas cells from another dog with B cell leukemia and both dogs with T cell leukemia expressed only mRNA for the beta and gammac subunits that comprise the intermediate-affinity receptor. These results indicate that the IL 2R is commonly expressed in canine lymphohematopoietic malignancies, and support the suitability of this large-animal model to evaluate targeted IL-2R cancer therapy using approaches of interest in the treatment of humans with hemolymphatic cancers. PMID- 11924910 TI - Enhanced anti-B-cell tumor effects with anti-CD20 superantibody. AB - The data presented here describe a novel approach to enhance the use of antibodies in diagnostic and therapeutic applications. Using a peptide copied from a rare self-binding (autophilic) antibody structure, the authors were able to convert by chemical cross-linking an anti-CD20 antibody to a self-binding (autophilic) structure. The autophilic antibody exhibited better binding to target tumor cells than the naked antibody. By the mechanism of hyper-cross linking a B-cell receptor (CD20) on tumor cells, the rate of apoptosis is significantly increased, leading to strong inhibition of tumor growth in culture. The demonstration of enhanced binding and apoptosis targeting the CD20 B-cell marker serves as an example for developing second-generation therapeutic antibodies against non-Hodgkin lymphoma. PMID- 11924911 TI - Identification of endogenous HLA-A2-restricted reactivity against shared melanoma antigens in patients using the quantitative real-time polymerase chain reaction. AB - This study was conducted to determine whether reactivity to melanoma cells of pretreatment peripheral blood mononuclear cells (PBMCs) from patients with metastatic melanoma correlated with subsequent response to treatment with interleukin-2 (IL-2). The sensitivity of the quantitative real-time polymerase chain reaction (PCR) assay was optimized, including the total number of cells used (3 x 10(6) in 1 mL), the responder-to-stimulator cell ratio (5:1), the optimal time to incubate PBMCs with tumor (2 h), the appropriate tumor stimulators (melanoma cell lines differing only in the expression of histocompatibility leukocyte antigen [HLA-A2]), the duration of recovery in the culture of PBMCs after cryopreservation (18-24 h), and the medium used (Iscove, 10% human AB serum). Using this optimized assay to detect HLA-A2-restricted antitumor reactivity in the pretreatment PBMCs from patients with melanoma, positive reactive responses were detected in 7 of 28 patients with an objective clinical response to IL-2 therapy compared with 6 of 21 positive reactive responses in nonresponding patients. None of 12 healthy donors were positive in this study. Thus, there was no significant difference in the reactivity of pretreatment PBMCs when responders were compared with nonresponders, although the melanoma patients had an increased incidence of response compared with healthy donors (p = 0.05). The PBMCs from 11 of the 13 melanoma patients with pretreatment HLA-A2-restricted antimelanoma reactivity were tested against a panel of transfectants expressing known shared melanoma antigens. Anti-MART-1 reactivity was detected in the pretreatment PBMCs of three patients. It thus appears that some melanoma patients are immunologically primed to antigens expressed on the tumor surface, although the HLA-A2-restricted antimelanoma activity detected in this real-time PCR assay was not predictive of patients' responses to IL-2 therapy. PMID- 11924912 TI - CD20-directed serotherapy in patients with multiple myeloma: biologic considerations and therapeutic applications. AB - Clonotypic B cells circulating in patients with multiple myeloma (MM) express CD20, and it has been suggested that these cells may be clonogenic. Furthermore, 20% of patients with MM express CD20 on their bone marrow plasma cells (BMPCs). Therefore, the authors began a phase II clinical study to determine the activity of the anti-CD20 monoclonal antibody rituximab in MM patients. Nineteen previously treated MM patients received 375 mg/m2 rituximab per week for 4 weeks. Three months after initiation of treatment, patients were assessed for response and received a second course of therapy if their disease was stable (SD) or they achieved a partial response (PR). Six of 19 (32%) patients had either a PR (n = 1) or SD (n = 5), with a median time to treatment failure of 5.5 months (mean, 10.3 months; range, 3-27+ months). All six patients who had a PR or SD had CD20+ BMPC. Overall, rituximab therapy was well tolerated. Because most patients with MM poorly express CD20 on their BMPCs, the authors evaluated agents for their ability to induce CD20 expression and thereby facilitate rituximab binding on MM cells. These studies show that interferon-gamma (IFN-y) induced CD20 expression on MM BMPCs, MM B cells, and healthy donor BMPCs. In contrast, CD20 expression on chronic lymphocytic leukemia, follicular non-Hodgkin's lymphoma, healthy donor B cells, and progenitor cells was unaffected by IFN-y. Rituximab binding to the BMPCs of MM patients was also increased after culture with pharmacologically attainable levels of IFN-gamma (1-100 U/mL). In conclusion, these studies suggest that MM patients with CD20+ BMPCs may benefit from rituximab therapy. Furthermore, IFN-gamma induces CD20 expression on MM BMPCs and B cells and facilitates rituximab binding to MM BMPCs, providing the rationale for clinical trials to examine its use with CD20-directed serotherapies in MM. PMID- 11924913 TI - Safety and efficacy of high-dose interleukin-2 therapy in patients with brain metastases. AB - The authors determined the safety and efficacy of recombinant high-dose interleukin-2 administration in patients with brain metastases. This retrospective review included 1,069 patients with metastatic melanoma or renal cell carcinoma who received high-dose interleukin-2 alone or in combination with other immunotherapy or chemotherapy from July 1985-July 2000. All patients were evaluated for both toxicity and response. Only the first exposure to interleukin 2 was considered. Parameters evaluated among the groups included toxicity profiles, reasons for stopping treatment, number of interleukin-2 doses per cycle, and response to therapy. Three patient groups were compared. Group I (n = 27) comprised patients with previously treated brain metastases (surgery or radiation), group 2 (n = 37) comprised patients with untreated brain metastases, and group 3 (n = 1,005) comprised patients without brain metastases. For most comparisons between patients with brain metastases and those without, no significant differences were noted in toxicity profiles or reasons for stopping interleukin-2 therapy. Patients with previously treated brain metastases received fewer interleukin-2 doses per cycle (median, 6.5) than patients with previously untreated brain metastases (median, 7.5) or patients without brain metastases (median, 7.5). Patients with previously treated brain metastases demonstrated an 18.5% overall clinical response to interleukin-2 treatment. However, patients with evaluable (previously untreated) brain metastases had an overall 5.6% response rate, which was less than the 19.8% response rate of patients without brain metastases. Two of thirty-six patients with evaluable brain metastases demonstrated objective regression of intracranial and extracranial disease after receiving interleukin-2. Carefully selected patients with brain metastases can safely receive high-dose interleukin-2, and some can experience a response to treatment at intracranial and extracranial disease sites. PMID- 11924914 TI - Identification of a clinical-grade maturation factor for dendritic cells. AB - Dendritic cells (DC) are essential for the generation of primary adaptive immune responses, but their full immunostimulatory capacities are only reached upon maturation. The authors compared several clinical-grade adjuvants of bacterial origin to determine their ability to induce phenotypic and functional maturation of monocyte-derived DC (Dendritophages, Dphi; IDM, Paris, France) differentiated with granulocyte-macrophage colony-stimulating factor and interleukin-13 in single-use cell processors (VacCell; IDM, Paris, France). Monophosphoryl lipid A, Mycobacterium bovis bacillus Calmette-Guerin, and Ribomunyl (Pierre Fabre Medicament, Boulogne, France) all appeared able to provide the signal necessary to initiate Dphi maturation. However, only Ribomunyl (Pierre Fabre Medicament) (containing membrane and ribosomal fractions from four bacterial strains) allowed the authors to obtain a significant enhancement of allostimulatory abilities and cytokine production by Dphi in the absence of active cellular infection. Addition of interferon-gamma (IFN-gamma) to Ribomunyl resulted in more pronounced upregulation of CD83, major histocompatibility complex class I, and B7 molecules by Dphi. Moreover, the IFN-gamma addition modulated their cytokine secretion, allowing higher levels of bioactive interleukin-12 concomitant with lower levels of interleukin-10. In kinetic studies, Dphi contact with Ribomunyl and IFN-gamma for 6 hours was sufficient to trigger a maturation process that completed spontaneously. Thus, Ribomunyl in association with IFN-gamma represents a suitable agent for the ex vivo production of mature monocyte-derived DC that can be used as cellular vaccines to promote a potent type I immune response. PMID- 11924915 TI - In utero transplantation of haemopoietic stem cells. AB - In utero haematopoietic stem cell transplantation is a potentially valuable therapeutic approach that strives to take advantage of biological opportunities for cellular transplantation that exist in the early gestational fetus. However, with the exception of severe combined immunodeficiency, clinical application has been limited by minimal or no engraftment, suggesting the presence of significant barriers to engraftment within the fetal environnment. Research directed toward elucidating these barriers is progressing, and there is hope that the barriers to engraftment can be overcome in the near future. In the meantime, there are a limited number of specific disorders that are biologically favourable and may be amenable to treatment by this approach using conventional techniques. In the future, strategies that improve the competitive capacity of donor cells or the use of pre-natal transplantation to induce donor-specific tolerance followed by post-natal non-myeloablative enhancement of donor chimerism may allow broad clinical application of this approach. PMID- 11924916 TI - Allogeneic transplantation using peripheral blood stem cells. AB - Over the past 9 years there has been a remarkable increase in the use of peripheral blood stem cells (PBSC) for allogeneic transplantation, primarily for matched sibling transplants but also increasingly for unrelated donor transplantation. In 1999 over 50% of all sibling transplants and over 25% of unrelated donor transplants reported to the European Group for Blood and Marrow Transplantation (EBMT) used PBSC. The major reason for this increasing use of PBSC relates to the rapid haemopoietic recovery seen which mirrors the advantages of using PBSC in autologous transplantation. This improvement in engraftment is a consequence of the larger number of stem cells that can be collected from G-CSF mobilized peripheral blood compared to bone marrow. Evidence from randomized trials now shows a survival advantage for the use of PBSC in patients with advanced leukaemia. The reason for this improved survival appears primarily to relate to a reduced risk of transplant-related mortality and, possibly, a reduced risk of relapse, However, these randomized studies have also confirmed that there is an increased risk of chronic graft-versus-host disease associated with PBSC transplantation and further follow-up is required to determine the long-term impact on outcome. PMID- 11924917 TI - Allogeneic transplantation using non-myeloablative transplant regimens. AB - Reduced intensity (non-myeloablative) stem cell transplant (NST) preparative regimens are being increasingly used to exploit the curative potential of allogeneic stem cell transplantation without the morbidity and mortality associated with conventional transplantation. Growing confidence in the power of the allogeneic graft-versus-malignancy (GVM) effect makes such an approach attractive. Lower intensity transplants increase the degree of mixed chimerism, both in T cell and myeloid cell lineages. Currently a variety of NST treatment approaches are being developed and in this chapter their safety profile and the immunological characteristics of the mixed chimeric state are described. Results of NST in specific disease categories are still limited but the NST approach appears to have promise in the treatment of both haematological and non haematological malignancies because of the benefit of low toxicity coupled with a strong graft-versus-malignancy effect. NST regimens are also being explored in high-risk patients with non-malignant disorders. However, at present, there is insufficient data to determine whether NST should replace standard myeloablative transplants in specific disease groups. With their low toxicity, NST are well placed as platforms for future developments in transplant immunology to avoid GVHD and enhance the allograft effect against malignant diseases. PMID- 11924918 TI - Exploiting graft-versus-tumour responses using donor leukocyte infusions. AB - Allogeneic stem cell transplantation (SCT) has become the treatment of choice for some patients with haematological malignancies, allowing dose escalation of chemo radiotherapy beyond the limits imposed by bone marrow toxicity. However, it is now apparent that dose escalation alone does not eradicate the malignancy in many cases and that an associated immune-mediated graft-versus-malignancy effect may be equally important. Its presence is supported by the following observations: anecdotal reports that patients with relapsed leukaemia following SCT may re enter remission after withdrawal of immunosuppressive drugs; the lower risk of relapse associated with the development of graft-versus-host-disease (GVHD); and an increased risk of relapse in patients receiving syngeneic transplants or T cell depleted allogeneic marrow grafts. More directly compelling evidence has been provided by the efficacy of donor lymphocyte infusions, particularly for relapsed chronic-phase CML. Issues that remain to be resolved include the precise nature of the effector cells and their target antigens, the best strategies for separating graft-versus-malignancy from GVHD, the role of adjuvant chemotherapy/cytokines, and the role of non-myeloablative transplantation. PMID- 11924919 TI - Allogeneic transplantation across major HLA barriers. AB - Only one-fourth of patients with fatal haematological disorders and malignancies will have an HLA-matched sibling donor to create access to potentially curative allogeneic stem cell transplantation. Advances in medical management, preparation of the graft and prevention of transplant-related complications, particularly rejection and GVHD, now make it possible to use alternative donors who are not genetically HLA-histocompatible with the patient. Some patients identify unrelated donor options using adult volunteers or cord blood units. All patients have immediate access to one or more genetically half-matched (haplo-identical), HLA partially-mismatched related donors. Using safer non-cytotoxic therapies to ablate the patient's immune system, graft preparation to decrease T-lymphocyte and/or increase CD 34+ cell doses, and post-transplant GVHD and infection prophylaxis, rates >95% for engraftment and <25% for grade II-IV GVHD can be achieved. Thus, disease-free survival rates are comparable to other alternative donors with all outcomes adversely influenced by advanced disease status, poor medical performance and older age. PMID- 11924920 TI - Stem cell transplantation for autoimmune diseases. AB - Much progress has been made in the field of haemopoietic stem cell transplants (HSCTs) for severe autoimmune disorders. Theoretical considerations, animal data and anecdotal evidence suggested some time ago that intensive immunoablation followed by autologous HSCT could restore normal immune reactivity in patients with severe autoimmune disorders. Based on a concept statement issued in 1995, two European societies, the European League Against Rheumatism (EULAR) and the European Group for Blood and Marrow Transplantation (EBMT) began collecting phase I/II trial data in an international collaborative network. Sufficient information from more than 350 patients allows a preliminary assessment with level three evidence. Autologous HSCTs can induce remissions in all disease categories tested so far. Remissions can be transient or durable. HSCTs are associated with significant morbidity and mortality. Treatment-related mortality (TRM) is near 10% at 1 year and is associated with the intensity of the conditioning and the stage of the disease at the time of transplant. Marked interdisease differences exist. There are few data available in haematological autoimmune diseases, more in systemic sclerosis (SSc), systemic lupus erythematosus (SLE), juvenile idiopathic arthritis (JIA) and multiple sclerosis (MS). Patient selection has been recognized as a crucial element from the phase I-II trials. Patients with advanced disease, severely compromised organ function or irreversible organ damage should not be considered as candidates for HSCT. Prospective randomized studies should now determine the value of HSCT compared to standard therapy. Such trials are ongoing for patients with systemic sclerosis (ASTIS trial--Autologous Stem Cell Transplantation International Scleroderma Trial) or are planned for patients with multiple sclerosis (ASTIMS trial--Autologous Stem Cell Transplantation International Multiple Sclerosis Trial) and rheumatoid arthritis (ASTIRA trial--Autologous Stem Cell Transplantation International Rheumatoid Arthritis Trial). More phase II data are needed for other indications such as SLE and JIA. PMID- 11924921 TI - Developments in the treatment of post-transplant viral disease. AB - Stem cell transplantation is a potentially curative procedure for many types of haemopoietic malignancy but is associated with a period of intense immunosuppression which may last for 1 to 2 years. During this time new viral infections or reactivation of endogenous, latent virus can lead to a variety of clinical symptoms which, together, make a significant contribution to the morbidity and mortality of the procedure. Improvements in viral detection, prophylaxis and treatment of established disease are helping to minimize the clinical impact of viral disease, but a definitive solution to this problem must await improvements in our ability to reconstitute an effective immune system. Each of the viruses implicated in post-transplant disease has a unique relationship with the host immune system, and there is every hope that improvements in our understanding of viral immunology will be translated into more effective clinical control. PMID- 11924922 TI - Allogeneic transplantation for leukaemia using unrelated donors. AB - This chapter describes the current role of unrelated donor stem cell transplantation (UD-SCT) in the management of leukaemia. The available data are scant and incomplete and there are few randomized studies comparing UD-SCT with alternative therapies. Patients with many of the leukaemias require prolonged follow-up after allogeneic SCT to determine whether they are cured; the registry based comparisons that have been initiated reflect the results achievable some years ago and may not help us in deciding what is best in 2001. In addition, new therapies such as ST1571, even though the long-term outcome of patients treated with this agent is uncertain, may affect which patients with chronic myeloid leukaemia we decide to recommend for transplant. The focus here is on acute and chronic myeloid leukaemia, acute lymphoblastic leukaemia and chronic lymphocytic leukaemia, as well as the myelodysplastic syndromes. Patient selection, conditioning strategies, comparison with other therapies, timing of transplant and the major causes of treatment failure are discussed, and there is an exploration of where improvement will come from. PMID- 11924923 TI - Allogeneic transplantation for haemoglobinopathies. AB - Beta-thalassaemia major and sickle-cell disease (SCD) reduce lifespan and quality of life for >300000 children and young adults worldwide. The only cure for both disorders is allogeneic stem cell transplantation (SCT). The decision-making processes in recommending SCT for patients with thalassaemia and SCD are different. For thalassaemia, where transfusion-related iron overload is universal, SCT should be offered to all patients <17 years because long-term survival and thalassaemia-free survival are about 80 and 70% respectively. For thalassaemics unable to comply with medical treatment, SCT offers a significant survival advantage; however, for patients with optimal medical care, short-term survival after SCT is inferior to medical treatment, and SCT instead offers a life free from transfusions and iron chelation. The clinical heterogeneity of SCD means that SCT is recommended only for selected patients with severe disease, particularly sickle-related neurological problems, for whom long-term survival and SCD-free survival after SCT approach 92 and 86% respectively. We here review the evidence available to help physicians evaluate the role of SCT for individual patients with thalassaemia major or SCD. PMID- 11924924 TI - Prospects for gene therapy using haemopoietic stem cells. AB - Gene therapy has thus far promised much and delivered little. Much of this has been due to deficiencies in the reagents and methodologies employed in early clinical trials. Recent technological advances in vectors and haemopoietic stem cell manipulation, coupled with improved pre-clinical assays of gene transfer and expression in re-populating stem cells give cause for greater optimism. Here we review these advances and indicate areas requiring further development before clinical gene therapy in the haemopoietic system becomes a widely applicable treatment modality. PMID- 11924925 TI - Telomeres and telomerase in human health and disease. AB - Epigenetics refers to the durable changes affecting the genome of an individual during development and aging, but which are not necessarily passed on to subsequent generations. Among the best studied of these epigenetic changes is the shortening of chromosome ends or telomeres. Telomeres are specialized structures, consisting of characteristic DNA repeat sequences and the complex of associated proteins, which cap and protect chromosome ends and serve to preserve genome integrity. In most somatic cells, progressive rounds of cell division are associated with telomere shortening. Such progressive attrition of telomere length eventuates in loss of replicative capacity (cellular senescence). In order to protect the germline and the subpopulation of stem cells from senescence, mechanisms have evolved to prevent telomere attrition in these cellular compartments. The most common and best studied mechanism involves the activation of a ribonucleoprotein enzyme complex, known as telomerase. Activity of telomerase circumvents loss of replicative capacity, by preserving telomere length and chromosome integrity. Hence the detailed mechanisms governing the expression and activity of telomerase have been intensively studied in development and differentiation. Early embryonic development and cellular differentiation are associated with a progressive diminution in telomerase activity. This decrease in activity is principally mediated at the level of the promoter for the gene encoding the catalytic unit of the telomerase complex. Unraveling the detailed mechanisms involved in the regulation of telomere length and telomerase activity will have important and far-reaching implications in understanding many aspects of human health and disease, ranging from accelerated aging syndromes to cancer pathogenesis, among others. Furthermore, insights gleaned from continuing research in this area will likely be applicable to the development of strategies to circumvent cellular senescence in regenerative medicine and stem cell therapeutics in the years to come. PMID- 11924926 TI - Monogenic forms of obesity and diabetes mellitus. AB - Great progress has been made in identifying several genes and in understanding the molecular pathogenesis of inherited syndromes of obesity and diabetes mellitus (DM). In humans, mutations in leptin, leptin receptor, proopiomelanocortin (POMC), melanocortin-4 receptor (MC4R) and prohormone convertase 1 (PC1) have been described in patients with severe obesity. Most of these obesity disorders, with the exception of the MC4R mutations, exhibit recessive inheritance and a distinct phenotype with varying degrees of hypothalamic dysfunction, and they unravel the critical role of the central leptin and melanocortin pathways in human appetite control and energy homeostasis. Maturity onset diabetes of the young (MODY) is a genetically and clinically heterogeneous subtype of type 2 DM with early onset autosomal dominant inheritance and a primary defect in insulin secretion. To date, six MODY genes have been identified, the glucokinase gene and five beta cell-specific transcription factor genes, hepatocyte nuclear factor-1alpha (HNF-1alpha), HNF 1beta, HNF-4alpha, insulin promoter factor-1 (IPF-1) and NeuroD1/BETA2. Mitochondrial DNA mutations cause another form of DM with an insulin secretory defect that is commonly associated with neurosensory hearing impairment, and has strict maternal inheritance. At the other end of the spectrum are the inherited syndromes of insulin resistance that are caused by mutations in the insulin receptor gene and in the adipocyte-specific transcription factor PPARgamma. The advances in our knowledge of the phenotypic manifestations and underlying molecular mechanisms of genetic syndromes of obesity and DM raise expectations for molecular diagnosis, as well as for more etiological therapies and better prevention of the continuously increasing prevalence of obesity and DM in our modern societies. PMID- 11924927 TI - Pseudohypoparathyroidism: a multitude of mutations in the stimulatory G protein alpha subunit (Gsalpha). PMID- 11924928 TI - Growth hormone binding protein, insulin-like growth factor-I and short stature in two pygmy populations from the Philippines. AB - The molecular basis and biochemical mediators of genetic growth propensity and adult height achievement in the general population are largely unknown. Pygmies represent one extreme of the height spectrum that may provide important clues regarding this issue. Previous studies in pygmies from Africa and Papua-New Guinea have shown decreased serum levels of growth hormone binding protein (GHBP), the circulating ectodomain of the growth hormone receptor (GHR). By inference, a similar limitation in tissue GHR expression has been assumed to be responsible for the partial growth hormone (GH) resistance observed in African pygmies. It is not clear how generalizable this concept is to other populations. To address this question, we studied two pygmy populations from the Philippines (Aeta and Mamanwa people) that are unrelated to the African pygmies. Serum GHBP and IGF-I levels were significantly decreased in both pygmy populations, compared to normal-statured Philippino controls. The results, together with previous observations in African and New Guinean pygmies, indicate that short stature is associated with low serum GHBP levels in pygmy populations of diverse origins and in different parts of the world. This strengthens the tentative postulate that the GHBP/GHR system plays an important role in the genetic and perhaps nutritional determination of adult stature in humans. Molecular genetic studies of the GHR gene in various pygmy populations may shed further light on the mystery of pygmy short stature. PMID- 11924929 TI - Psychomotor development in the children of mothers with type 1 diabetes mellitus or gestational diabetes mellitus. AB - The purpose of this study was to examine psychomotor development in children born to mothers with type 1 diabetes mellitus (DM1) or gestational diabetes mellitus (GDM). The influence of metabolic control in pregnant diabetic mothers and complications during labor on their children's psychological and physical development was evaluated. The analysis included 59 children, 20 of mothers with GDM, 19 of mothers with DM1, and 20 children of healthy mothers. Clinical observations and medical history were recorded and children were assessed using the Brunet-Lezine Psychomotor Development Scale. Abnormalities were found more often in the children of mothers with DM1 whose illness was insufficiently controlled during pregnancy and of mothers with serious hypoglycemia while pregnant. Speech, eye-movement coordination and social aspects were affected. PMID- 11924930 TI - Clinical characteristics of non-immune-mediated, idiopathic type 1 (type 1B) diabetes mellitus in Japanese children and adolescents. AB - To clarify the characteristics of idiopathic type 1 (type 1B) diabetes mellitus (DM), we compared the clinical features of immune-mediated type 1 (type 1A) DM and type 1B DM in 85 Japanese children and adolescents with DM. The prevalence of type 1B DM was 16.5%. The patients with type 1B DM were significantly younger at diagnosis and had a higher frequency of preceding viral infection before onset, compared to those with type 1A DM. They displayed more severe metabolic decompensation with a higher frequency of ketoacidosis at diagnosis than patients with type 1A DM. They had strong, HLA-defined genetic susceptibility, similar to that in type 1A DM. Some patients with type 1B DM exhibited a remarkably abrupt onset and rapid loss of beta-cell capacity. From these findings, it is considered that type 1B DM differs from type 1A DM with respect to age at onset and the trigger event, such as viral infection, leading to rapid destruction of beta cells without autoimmunity in the etiology of the disease. PMID- 11924931 TI - Hypothalamic-pituitary-testicular function in prepubertal children with chronic liver disease. AB - Adult patients with chronic liver disease (CLD) show clinical and biochemical signs of hypogonadism and estrogenization. However, no information is available on hypothalamo-pituitary-testicular function in prepubertal or early pubertal children with CLD. Eighteen prepubertal children with CLD, aged 5.8+/-5.5 years (mean +/- SD; range 0.32-12.8), were studied. Most of them had moderate liver function abnormality. Height was slightly decreased (SDS: -1.44-/+1.88) but weight for height was adequate. Serum gonadotropins were evaluated as a function of age. In the age group younger than 1 year (n = 7), serum LH was elevated (4.88+/-6.22 IU/l) when compared with a group of 39 control children (1.2+/ 1.65), while serum FSH was normal. In this young group, serum testosterone was normal, but serum estradiol was significantly increased (24.1+/-19.7 pg/ml) when compared with the control group (6.5+/-3.54). In contrast, in the age group older than 2 years, no difference between patients with CLD and controls was observed, either in serum gonadotropins or in serum sex hormones. Taking the 18 patients with CLD together, serum SHBG (113.7+/-51 nmol/l; mean +/- SD) was significantly higher than in normal controls (76+/-38 nmol/l, n = 91, p <0.001). Moreover, and different from normal controls, no change with age was observed in serum SHBG, total testosterone or bioavailable testosterone (non-SHBG-bound). Normal testosterone response to hCG stimulation (>1 ng/ml) was found in a subgroup of 11 children with CLD. By contrast, eight of 11 patients with CLD had an inadequate decrease in SHBG after androgen stimulation. In conclusion, we observed that during the first year of life, a period which includes the postnatal activation of the hypothalamo-pituitary-testicular axis, there is an elevation of serum LH and serum estradiol that suggests the existence of a moderate deficiency of Leydig cell function. This disorder is no longer observed in older prepubertal children. Similar to reports in adults, children with CLD have elevation of serum SHBG levels. Furthermore, the lack of SHBG decrease and bioavailable testosterone increase with age, probably modulated by GH, suggests some degree of hepatic GH resistance in prepubertal patients with CLD. PMID- 11924932 TI - Adult height in advanced puberty with or without gonadotropin hormone releasing hormone analog treatment. AB - Advanced puberty is defined as the onset of puberty in girls at 8-10 years of age and in boys at 9-11 years. This study analyzes adult height in 57 children with advanced puberty to evaluate the results of treating children (9 girls and 8 boys) with gonadotropin hormone releasing hormone (GnRH) analog and the impact of advanced puberty on adult height in untreated children (31 girls and 9 boys). For treated girls, adult height predicted at the onset of treatment (151.9+/-1.7 cm) was similar to the final adult height (155.3+/-1.4 cm), but lower than target height (157.2+/-1.6 cm, p = 0.04). For untreated girls, adult height predicted at the initial evaluation (156.7+/-1 cm) was also similar to adult height (157+/-1 cm), but lower than the target height (157.6+/-1 cm, p = 0.03). The adult heights of both treated and untreated girls were similar to their target heights. For treated boys, adult height predicted at the onset of treatment (173.2+/-3.1 cm) was greater than the final adult height (164.1+/-2.1 cm, p = 0.01), which was lower than target height (170.4+/-1.2 cm, p = 0.01). For untreated boys, adult height predicted at the initial evaluation (170.8+/-2.7 cm) was similar to both the adult height (169.1+/-1.9 cm) and target height (170.2+/-1.2 cm). Height gains between the onset of puberty and adult height were similar in treated (29.9+/-2.3 cm in girls and 29.8+/-1.7 cm in boys) and untreated (28.6+/-1 and 33.1+/-2 cm) children. When expressed as SD, the adult height was significantly shorter than that at 4 years in treated girls (difference 1 SD, p = 0.03), in untreated girls (difference 0.9 SD, p = 0.0002) and in treated boys (difference 0.9 SD, p = 0.02), but it was similar to that in untreated boys. Adult height was below target height by >5 cm in seven girls (two of them treated) and five boys (four of them treated). In conclusion, treating advanced puberty did not change the adult height reached by girls, and was associated with reduced growth potential in boys. The adult heights of untreated children were similar to those predicted at the initial evaluation and to target heights, but in girls they were 1 SD lower than the height at 4 years. These data suggest that advanced puberty decreases the growth potential by about 5 cm, and that GnRH analog treatment does not prevent this. PMID- 11924933 TI - The relation between adrenal function and the severity of bronchial hyperresponsiveness in children as measured by the methacholine provocation test. AB - BACKGROUND: There is no satisfactory explanation why some individuals experience severe attacks of asthma, yet others, exposed to similar stimuli, have a milder form of the disease. OBJECTIVE: We tested the hypothesis that children with more severe disease may have relative adrenal insufficiency compared to the children with milder disease. PATIENTS AND METHODS: Sixteen children with chronic asthma aged 8-16 years old were studied. Adrenal function was evaluated by the 24-h excretion of urinary free cortisol (UFC) before and after ACTH stimulation, and by plasma cortisol levels before and 60 min after ACTH administration. The severity of bronchial hyperresponsiveness was evaluated by the methacholine provocation test. RESULTS: Nine children had 20% fall in forced expiratory volume in 1 sec (FEV1) after a provocative concentration (PC20FEV1) of methacholine > or =2.5 mg/ml and were considered as having mild-moderate bronchial hyperresponsiveness (Group A). Seven children had a PC20FEV1 of < or =1.25 mg/ml and were considered as having severe bronchial hyperresponsiveness (Group B). No significant difference was found between the peak plasma cortisol response to ACTH between the two groups (634+/-182 and 586+/-137 nmol/l, respectively). However, there was a significant statistical difference (p <0.01) in the 24-h UFC response to ACTH between the children from Group A (345+/-107 nmol/m2 ) and the children from Group B (161+/-125 nmol/m2). CONCLUSIONS: Based on the low levels of 24-h UFC secretion in severely asthmatic children in our study, we propose the encouragement of provision of a short course of inhaled steroids to be kept at home for the emergency therapy of those children identified as having high-risk asthma. PMID- 11924934 TI - Diurnal leptin rhythms in children treated with prednisolone once daily in the morning or in the evening. AB - Serum leptin levels exhibit a marked diurnal variation in children. The aim of the present study was to investigate whether the timing of administration of exogenous glucocorticoids, which have been found to increase serum leptin, affects the diurnal rhythm. Four girls and four boys aged 10.6 to 15.8 (mean 13.2) years with asthma were studied. The design was an open 2-period cross-over trial with a 1-day run in, two 4-day periods of prednisolone 5 mg in the morning or in the evening, with a 3-week washout in between treatment periods. During run in and on the last day of the prednisolone periods a fasting blood sample was drawn at 08.00 h, and thereafter samples were obtained every 2 h throughout the day until 08.00 h the next morning. Serum leptin was measured by a specific radioimmunoassay. During all periods, leptin levels were low during the day with a nadir at 10.00 h (run-in [mean +/- SEM]: 3.9+/-1.28; morning prednisolone: 5.2+/-1.58; evening prednisolone: 5.7+/-2.02 microg/l). Increases in leptin levels were detected from 20.00 h with zeniths at 24.00 h (run in: 7.2+/-1.86; evening prednisolone: 9.2+/-2.36 microg/l) and 02.00 h (morning prednisolone: 9.4+/-1.78 microg/l) (F = 115.5; p <0.01). As compared to run in, leptin levels were increased at all time points during prednisolone treatment in the morning (F = 16.0, p = 0.01) and in the evening (F = 12.6, p = 0.01). No statistically significant differences were found in leptin levels during prednisolone in the morning or in the evening (F = 0.44, p = 0.53). Therefore, the timing of administration of exogenous glucocorticoids does not affect diurnal leptin rhythms in children. PMID- 11924935 TI - Transient neonatal hypoglycemia--long-term effects on neurodevelopmental outcome. AB - OBJECTIVE: To investigate the frequency, etiology and consequences of neonatal hypoglycemia. STUDY DESIGN: Ninety-four infants admitted to Gazi University Hospital neonatal intensive care unit for hypoglycemia (blood glucose <2.2 mmol/l 140 mg/dl]) over the past 5 years were identified and investigated with regard to cause, duration of treatment and neurological outcome. RESULTS: The frequency of neonatal hypoglycemia in our unit was 94/1,023 (9.18%). Twelve infants with hypoglycemia were small for gestational age (SGA), 55 were appropriate for gestational age (AGA), and 27 were large for gestational age (LGA). The cause of the hypoglycemia was not identified in 53 infants. SGA infants required the longest duration of i.v. glucose infusion. Forty-eight patients received psychometric evaluation, one patient showed a language deficit and two patients showed motor deficits. CONCLUSION: Neonatal hypoglycemia is a dangerous condition for its acute and chronic complications, and may be observed in infants with no clear risk factors. However, if acted upon early, these complications are preventable with mostly very simple measures. PMID- 11924936 TI - A novel mutation in PIT-1: phenotypic variability in familial combined pituitary hormone deficiencies. AB - Mutations in PIT-1 have been described in several cases of familial combined pituitary hormone deficiencies. This study describes a novel PIT-1 mutation that was found in two siblings of a highly consanguineous family of Israeli-Arab origin. The missense mutation (G688A) causes a lysine (K) to glutamic acid (E) substitution at codon 230. This codon resides in the first helix of the POU homeodomain, which is directly involved in DNA binding. This amino acid is conserved in most homeodomain proteins, suggesting that the substitution disrupts the DNA-binding activity of the mutant protein. Two main observations are described: 1. The clinical presentation of the mutation involves intrauterine growth retardation. 2. One sibling had full deficency of growth hormone and thyroid stimulating hormone, whereas the other had only growth hormone deficiency. This is, to the best of our knowledge, a unique expression of a novel PIT-1 mutation. PMID- 11924937 TI - The DAFNE initiative: the methodology for assessing dietary patterns across Europe using household budget survey data. AB - OBJECTIVE: The DAta Food NEtworking (DAFNE) initiative was conceived in the 1980s and aims at using data already collected in the context of household budget surveys (HBSs) for the assessment of dietary patterns across Europe. DESIGN: HBSs are country-representative surveys conducted, with similar methodology, at regular time intervals by the National Statistical Offices of almost all European countries. DAFNE focuses at the designation of comparable, between European countries, categories of food and socio-demographic data, and the estimation of daily per capita food availability. With food and socio-demographic data classified into common categories, comparisons of food habits between European countries become possible. SETTING: Fifteen European countries with one or more HBS all undertaken during the period 1981-1999. SUBJECTS: Representative population samples in 15 European countries. RESULTS: DAFNE HBS data are comparable and allow a wide range of dietary analyses. Preliminary data document differences in food habits among European countries and provide insights into the socio-demographic determinants of food preferences. CONCLUSION: The DAFNE databank, if properly expanded and exploited, could become a valuable tool for national food and nutrition policy planning across Europe and for the identification of groups at higher risk for developing nutrition-related conditions. PMID- 11924938 TI - Methodological issues using household budget survey expenditure data for individual food availability estimation: Irish experience in the DAFNE pan European project. DAta Food NEtworking. AB - OBJECTIVE: Irish participation in the EU-supported DAta Food NEtworking (DAFNE) project required compliance with the overall aims and objectives. The Irish Household Budget Survey (HBS) expenditure data had to be transformed into a format compatible with the collaborative effort, by converting them into quantities of foodstuffs available per person per day. SETTING: The Irish 1987 HBS expenditure data on all commodities for 7705 households in the Republic of Ireland, collected using a 14-day diary kept by all members of the household aged 15 years and over. DESIGN: Following identification of 188 food items in the HBS dataset, retail prices per unit weight were sought for each food. Adjustment of prices, collected from a number of different sources, was made to those of 1987 using the Consumer Price Index. Simple models were used to estimate household food availability through application of the adjusted retail prices per unit weight to the expenditure data. The household level data were converted to food availability per person per day. An internal validation of quantities estimated using the retail prices was made using the 12 foodstuffs for which the Irish HBS collects expenses and quantities. RESULTS: The comparison of quantities published by the Irish Central Statistics Office for 12 foodstuffs in the Irish 1987 Household Budget Survey with the quantities estimated using equivalent expenditure data and corresponding retail prices showed agreement, with less than a 10% margin of error for 10 of the foods. CONCLUSION: In spite some difficulty in converting HBS food expenditure data into food availability per person per day, the DAFNE approach is potentially useful for Irish nutrition surveillance purposes and for facilitating comparisons of the Irish HBS food data with those of other European countries. PMID- 11924939 TI - Estimation of age- and gender-specific food availability from household budget survey data. AB - OBJECTIVE: To derive estimates of age-gender specific food availability, based on data collected at household level. DESIGN: Two alternative modelling approaches are described leading to linear and non-linear optimisation, respectively. The idea of penalised least squares is used for estimation of model parameters. The effect of household characteristics can be incorporated into both modelling approaches. SETTING: Household budget survey data from four European countries (Belgium, Greece, Norway and the United Kingdom), circa 1990. PMID- 11924940 TI - Methodology for rendering household budget and individual nutrition surveys comparable, at the level of the dietary information collected. AB - OBJECTIVE: To describe the methodology applied in order to render comparable, at the level of the dietary information collected, the household budget survey (HBS) and individual nutrition survey (INS) data from four European countries (Belgium, Greece, Norway and the United Kingdom). SETTING: In Belgium, data from the HBS of 1987-88 were compared with data from the Belgian Interuniversity Research on Nutrition and Health collected from 1980 to 1985. In Greece, data from the HBS undertaken in 1993-94 in the greater Athens area were compared with data collected around 1994 in the same region, in the context of the Greek segment of the European Prospective Investigation on Cancer and Nutrition study. In Norway, data from the HBS carried out in 1992, 1993 and 1994 were compared with the NORKOST study conducted in 1993-94. In the United Kingdom, data from four HBSs carried out in 1985, 1986, 1987 and 1988 were compared with the National Dietary and Nutritional Survey of British adults conducted in 1987-88. DESIGN: INS generated data were converted into 'HBS-like' estimates with the application of yield factors for weight changes during cooking, recipe-based calculations and edible proportion coefficients taking into account weight changes during the food preparation. The 'HBS-like' estimates thus obtained were compared with the original HBS values, after applying an adjustment factor for food spoiled or given to pets. CONCLUSION: The methodological considerations overviewed in the present paper indicate that a number of issues need to be taken into account before a proper comparison of the dietary data collected through surveys implemented with varied methodologies is carried out. PMID- 11924941 TI - A preliminary assessment of the use of household budget survey data for the prediction of individual food consumption. AB - OBJECTIVE: To compare individualised household budget survey (HBS) data with food consumption values derived from individual nutrition surveys (INSs). SETTING: Four European countries: Belgium, 1980-85 and 1987-88; Greece, 1993-94; Norway, 1992-94; and the United Kingdom, 1985-88. DESIGN: Household budget survey data were individualised with non-parametric models. Individual nutrition survey data were converted into 'HBS-like' estimates, with the application of recipe-based calculations and yield factors for weight changes during food preparation. To correct for over- and underreporting or recording in both surveys, quantities (in g day(-1)) of 14 principal food groups were expressed as fractions of total food quantity (in g day(-1)). For each food group, INS and HBS-derived mean values were calculated for 24 research units, jointly defined by country (four countries involved), gender (male, female) and age (younger, middle-aged and older). Pearson correlation coefficients were calculated and correlation diagrams were drawn. CONCLUSION: The results of this preliminary analysis show that there is value in the nutritional information derived from HBSs. However, additional and more sophisticated work is required, in order to derive reliable point and interval estimates of individual food consumption based on HBS data. PMID- 11924942 TI - Portuguese household food availability in 1990 and 1995. AB - OBJECTIVE: To examine the changes in Portuguese household food availability from 1990 to 1995. DESIGN: Using the DAta Food NEtworking (DAFNE) classification scheme for nutritional and socio-economic variables, data from the Portuguese Household Budget Surveys collected in 1989/90 and 1994/95 were analysed. The mean availability (quantity person(-1) day(-1)) for the total population, by locality and by level of education of the household head, was computed for each of the main food and beverage groups. SETTING: Portugal, 1989 to 1995. RESULTS: Although still among one of the countries with high availability of cereals, pulses, fish and seafood and olive oil, Portugal is gradually moving away from the traditional 'Mediterranean diet'. Between 1989 and 1995, the availability of complex carbohydrates and olive oil was reduced, while the availability of protein supplying food groups increased. Considerable disparities can be observed by locality and by the level of education of the household head. Households in urban areas and of higher socio-economic status are the main actors of the changes in 'traditional' food habits. CONCLUSION: Although these findings are based on household food availability rather than consumption, they indicate the direction of the changes taking place in the Portuguese diet and can be used effectively by agriculture specialists, nutrition experts and policy makers. PMID- 11924943 TI - An overview of food eaten outside the home in the United Kingdom National Food Survey and the new Expenditure and Food Survey. AB - OBJECTIVE: To summarise the methods of data collection on foods eaten outside the home in the National Food Survey (NFS) and the new (2001) Expenditure and Food Survey (EFS). DESIGN: The treatment of foods eaten outside the home in the NFS was reviewed as part of a merger process with the UK household budget Family Expenditure Survey (FES). Problem issues were identified and the way these problems are being dealt with in the EFS is indicated. SETTING: United Kingdom. RESULTS: Pilot EFS results indicate improved coverage of foods eaten outside the home, in a comparable period to the NFS. CONCLUSION: The new EFS, which represents a merger of the NFS and the FES at the level of data collection and validation, is likely to preserve the qualities of the previous surveys and represents an improvement over them. PMID- 11924944 TI - Comparability of household and individual food consumption data--evidence from Sweden. AB - OBJECTIVE: Comparison of household and individual food consumption. DESIGN, SETTING AND SUBJECTS: Combined household and individual food consumption survey carried out in Sweden in 1989. A random sample of 3000 subjects aged 0-74 years, the household to which the subject belonged constituted the household unit. Each household recorded all the foods it purchased over a 4-week period, except food eaten outside the home. For the selected subject, excluding children <1 year old, food intake was obtained with a simplified 7-day record. Food consumption from the two data sets was compared for the whole sample and for one-person households, respectively. RESULTS: Reasonable agreement (+/-20%) was seen for many major foods including cereal products, milk, cheese, meat and meat products, and fish, and after correction for inedible part, fruit and vegetables. Purchases were lower for sweet bakery products, alcoholic beverages and potatoes, while the opposite was seen for oils and fats, cream and sugar. For one-person households the two data sets showed similar differences in consumption patterns between men and women. The gross differences observed can be explained by factors like home baking, eating out and recorded level of processing, e.g. as raw food including inedible part or as a prepared dish. CONCLUSIONS: The results show reasonable agreement between purchased and eaten amounts for aggregated food groups but marked differences were seen for some important individual items. Household-based consumption data are useful for many purposes provided the limitations of the data are accounted for. PMID- 11924946 TI - The DAFNE databank as a simple tool for nutrition policy. DAta Food NEtworking. AB - OBJECTIVE: To identify, quantify and depict variation of food habits in Europe, using data from the DAFNE (DAta Food NEtworking) databank. SETTING: Household budget survey data of 12 European countries, namely Belgium, Germany, Greece, Hungary, Ireland, Italy, Luxembourg, Norway, Poland, Portugal, Spain and the United Kingdom, around 1990. RESULTS: Data from the DAFNE databank are presented in simple pictorial presentations, which reveal considerable disparities in food habits. Furthermore, there appears to be nutritional variation within countries by socio-demographic groups, defined by their residence and educational level. The distribution patterns of food availability provide insights into the determinants of food preferences, as conditioned by current forces. CONCLUSION: The factors influencing consumer choice are many and varied. Thus, in order to promote healthy eating, it is essential to identify the food habits of the target population. In this context, information derived from household budget surveys, used in the development of the DAFNE databank, could be very important. PMID- 11924945 TI - The Household Food Consumption and Anthropometric Survey in Poland. AB - OBJECTIVE: To provide an overview of the Household Food Consumption and Anthropometric Survey, undertaken in Poland from September to November 2000. DESIGN: A sub-sample of households participating in the countrywide, representative household budget survey (HBS) was selected to participate in the Household Food Consumption and Anthropometric Survey. Four thousand two hundred (4200) individuals provided 24-hour recalls that were subsequently evaluated. Body weight and height together with the mid-arm, waist and hip circumferences were measured. Laboratory analyses of contaminants that may be present in selected individual diets will be undertaken. SETTING: Poland, 2000. SUBJECTS: Four thousand two hundred individuals from 1362 households participating in the Polish HBS. RESULTS: Preliminary analyses of height and weight, as well as energy and nutrient intakes, were undertaken in a sub-sample of 484 boys and girls aged 10-15 years. Energy intakes were generally in line with or above the Polish Recommendations. CONCLUSIONS: Apart from serving as an important information source on individual food consumption and anthropometric status of the Polish population, the data collected through the Household Food Consumption and Anthropometric Survey will allow direct comparisons of food intake estimates, based on 24-hour recalls, with the results of the household budget survey. Such comparisons are expected to improve and refine interpretation of the data derived from both the individual nutrition survey and the household budget survey, and particularly to provide information on the importance of eating out, which is not covered in the Polish HBS. PMID- 11924948 TI - Endocarditis risks. PMID- 11924947 TI - Corporately speaking. PMID- 11924949 TI - Occlusal considerations. PMID- 11924950 TI - Paracetamol pain relief. PMID- 11924951 TI - Stimulating new bone repair therapy. PMID- 11924952 TI - Teething troubles? AB - The relationship between the eruption of the deciduous teeth and the general health of infants has been documented for over 5,000 years. A variety of physical disturbances (anything from minor upsets to potentially fatal illnesses) have historically been attributed to teething, however a number of recent publications have alluded to a clarification of some of the disputed features of teething. It is now accepted that the localised symptoms of teething vary between individuals, however, 'teething' continues to be an inappropriate diagnosis proffered by both healthcare professionals and lay people. Severe systemic upsets are unrelated to teething and, if present, the infant should be promptly referred to a physician for an accurate diagnosis and appropriate treatment. The treatment modalities used in teething have been diverse throughout the ages, frequently depending on the tenets of the medical profession and lay people, but now principally involve pain relief. This article examines the signs and symptoms frequently attributed to teething and their possible alternative causes. The contemporary principles of the management of teething are discussed, including supportive measures, the diverse range of available topical and systemic pharmacological preparations and the 'alternative' holistic therapies. PMID- 11924953 TI - Crowns and extra-coronal restorations: considerations when planning treatment. AB - Considerations when planning treatment is the third in the series of crowns and other extra-coronal restorations. Articles or chapters on treatment planning in restorative dentistry can make pretty dry reading, often built around a list of factors that might influence your decision-making. In truth though, planning and placing crowns or other extra-coronal restorations cannot be distilled into a series of lists. The decision-making involved requires experience, subtle understanding and a flexible approach, none of which come easily. PMID- 11924954 TI - Screening panoramic radiography of new adult patients: diagnostic yield when combined with bitewing radiography and identification of selection criteria. AB - OBJECTIVES: To measure the radiological diagnostic yield on screening panoramic radiographs taken of new adult patients and to identify selection criteria for panoramic radiography of new adult patients. DESIGN: Survey of 1,817 consecutive panoramic radiographs taken as 'routine' on new patients with statistical analysis of clinical and radiological findings. SETTING: All radiographs were obtained from 41 general dental practitioners (GDPs). The GDPs provided the clinical information about the patient obtained by history and examination. Collection of material occurred in 1998/1999. MATERIALS AND METHODS: Two dental radiologists recorded the radiological findings on each of 1,817 panoramic radiographs by consensus. Those findings that would have been identified from bilateral posterior bitewing radiographs of each patient were then excluded to give modified figures for radiological findings. MAIN OUTCOME MEASURES: Indices of diagnostic yield were devised and calculated for each radiograph from the data on radiological findings. Total diagnostic yield (DY) and modified diagnostic yield (MDY), after exclusion of findings identifiable on bitewing radiographs, were both calculated. Clinical indicators of a high MDY were identified using stepwise multiple regression analysis. RESULTS: MDY was 0 for 17% of the radiographs (all patients) and 23% of the radiographs when the asymptomatic group were considered. The clinical variables for which the significance was high (p < 0.001) were: increasing number of teeth with clinical suspicion of periapical pathology, presence of partially erupted teeth, increasing number of clinically evident carious lesions, partially dentate status and presence of crowns. CONCLUSION: Taking posterior bitewing radiographs of new adult patients would reduce the diagnostic yield identified solely by panoramic radiography. Using clinical factors derived from the history and examination as radiographic selection criteria modestly improves the odds of achieving a high diagnostic yield from panoramic radiography. PMID- 11924957 TI - A low dispersion geometry for microchip separation devices. AB - Curved channel geometries introduced on microchip separation devices to achieve greater separation distances often lead to large analyte dispersion, degrading the performance of these systems. While such electrokinetic dispersion may be minimized by reducing the channel width around the curved region, alternative strategies involving larger channel curvatures may be promising as well, depending on the application. For example, Culbertson et al. (Anal. Chem. 2000, 72, 5814-5819) recently demonstrated the effectiveness of gentle spiral geometries in carrying out separations of small molecules. For moderate and large Peclet number systems, however, larger spiral geometries are necessary to diminish electrokinetic dispersion of solute slugs which may not conform to the needs of the microchip format. In this work, we investigate a modified spiral geometry with a wavy wall along the inner track of the channel. Analysis shows that such width profiling may significantly improve the performance of the spiral geometry, making the design effective for larger Peclet number or smaller radii systems. Numerical simulations performed to optimize these modified spirals suggest equating transit times along the inner and the outer track of the channel as a useful design criterion for minimizing electrokinetic dispersion. An analytical model has been formulated to derive the optimal channel parameters based on this criteria which compares well with the simulation results. PMID- 11924955 TI - Dental pain and dental treatment of young children attending the general dental service. AB - OBJECTIVES: The objective was to examine the relationship between dental pain (and its sequelae), and the extent of restorative care provided for primary molars, amongst children who regularly attend a general dental practitioner. METHODS: A retrospective review of the clinical case notes of 677 children with caries who attended 50 general dental practitioners on a regular basis. Analyses were performed at the subject level. Logistic regression models were fitted for the dependent variables whether or not pain, a dental extraction for pain or sepsis and a course of antibiotics was recorded, after taking into account the proportion of carious teeth that were restored, the total number of carious teeth, the age caries was first recorded, gender and the clustering of the subjects within dental practices. RESULTS: Almost half of the children in the study (48%) were recorded as having at least one episode of pain. Total decay experience in the primary molars was a significant predictor of pain, extraction due to pain or sepsis and prescription of antibiotics. There was no significant association between the proportion of carious teeth restored and each of the three dependent variables. CONCLUSIONS: For those children who regularly attend their general dental practitioner and who have decay in their primary molars, dental pain is a common finding. Total decay experience in primary molars is the principal predictor of pain, extraction due to pain and the need for antibiotics, whilst the level of restorative care in the primary dentition is less important. In order to reduce the incidence of dental pain in young children, effective methods of preventing caries at the individual and public health levels need to be expanded. PMID- 11924956 TI - Quality assurance by service quality--an industrial approach to monitoring course delivery. AB - OBJECTIVE: To evaluate the use of an adapted customer service quality questionnaire as a management tool in the delivery of an undergraduate phantom head course. DESIGN: Two questionnaires, based upon an industrial definition of quality service, were devised and distributed for completion by second year dental students attending the management of dental caries course. These were applied to the academic years 1997/98 and 1999/2000. The first questionnaire was prospective, administered prior to course commencement, and sought to ascertain the level of importance attached to each dimension of course delivery. The second was retrospective and measured the level of satisfaction with course delivery. By comparing the 1997/98 responses for both questionnaire types, areas requiring improvement were identified. Where these areas were emphasised prospectively as important by the 1999/2000 students' improvement measures were introduced. Their effect was assessed by the subsequent retrospective satisfaction questionnaire. RESULTS: In all cases the questionnaire return rate was 100%. Although across the academic years overall student expectations did not differ significantly (P > 0.05) the emphasis of different components was found to vary according to academic year. CONCLUSION: The application of this technique assists the course organiser to make objective decisions on improving course delivery based upon the shortfalls of the previous year and the pre-course expectations of the following academic year. The industrial model thus translated well to an educational context and should prove useful in continually improving course delivery. PMID- 11924958 TI - A study of the critical criteria for analyte stability in high-temperature liquid chromatography. AB - There are three major impediments to the use of high-temperature ultrafast liquid chromatography. First, the stationary phase must be thermally stable. Over the past decade, a series of thermally stable, highly efficient stationary phases have been developed that can withstand temperatures exceeding 200 degrees C. Second, the temperature mismatch between the incoming eluent and the column must be minimized (<5 degrees C), because such a mismatch is a very serious cause of peak broadening, especially in ultrafast separations. The thermal mismatch problem can be significantly ameliorated at high column linear velocities by using narrow-bore columns (2.1-mm i.d.). Third, analytes that are exposed to high temperatures must be thermally stable on the time scale of the chromatographic run. We report here a study of the ability of a number of pharmaceuticals to withstand superambient temperatures on the time scale of fast separations. We propose criteria by which a particular analyte may be rejected as a candidate for high-temperature analysis, and we demonstrate that complex molecules are amenable to quantitation, even at temperatures in excess of 100 degrees C in the aqueous media. We also show that as the time an analyte spends on hot column decreases, the extent of on-column reaction decreases for those analytes that do react. Although the seminal work of Antia and Horvath addresses these issues from a theoretical perspective, we hope to further alleviate fear of the use of high temperatures in liquid chromatography through the empirical approach used here. PMID- 11924959 TI - Geometric scaling effects in electrical field flow fractionation. 2. Experimental results. AB - Geometric scaling of microelectrical field flow fractionation (micro-EFFF) systems is investigated experimentally and compared to theory and to macroscale EFFF systems. Experimental results are presented to demonstrate that the miniaturized system operates according to the scaling theory associated with the system. Demonstrated improvements in the channels include increased retention and resolution and decreased peak broadening, electrical time constants, relaxation time, power consumption, and sample size. Additionally, scaling effects related to the compression of separation zones in the miniaturized EFFF systems are discussed. PMID- 11924960 TI - Extending the solid-phase microextraction technique to high analyte concentrations: measurements and thermodynamic analysis. AB - The solid-phase microextraction (SPME) technique has been used historically to quantify analytes present at the parts per million level. However, the nonintrusive nature of SPME lends itself to other applications involving analytes at higher concentration. In the current work, the possibility of using the SPME technique to measure concentrated gaseous samples was examined. Pentane concentrations between 0 and 100% saturation were studied, over a temperature range of 20-45 degrees C. The results showed that, up to a critical mole fraction in the solid phase, the concentrations of pentane in the polymeric extracting solid and vapor phases were related by a constant, equal to Henry's constant. The temperature dependence of Henry's constant was shown to follow the predicted trend with temperature, as determined from rigorous thermodynamic calculations. Above the pentane concentration in the polymeric phase, the response deviated from linearity. The nonideality was captured in an activity coefficient. An activity coefficient model developed to describe the nonideality was found to be a function of the swollen volume of the SPME polymer phase. The results indicate that the SPME technique can be applied to high analyte concentrations, although difficulties may be encountered when multiple analytes are absorbed. PMID- 11924961 TI - Chromatofocusing using micropellicular column packings with computer-aided design of the elution buffer composition. AB - Micropellicular, anion-exchange column packings are used in chromatofocusing to demonstrate the resolution and speed achieved when proteins are separated under these conditions. Linear or concave pH gradients are produced with simple mixtures containing four or fewer individual buffering species instead of the more commonly used polyampholyte buffers. Computer-aided design methods are demonstrated for selecting the composition of the elution buffer to produce a pH gradient of a desired shape. The method is applied to high-resolution, analytical and preparative-scale separations involving horse myoglobin, human hemoglobin variants, and bovine carbonic anhydrase. A useful selection of buffering species is described capable of producing pH gradients of a variety of shapes in the range between pH 9.5 and 5.5. PMID- 11924962 TI - Fluorescence-labeled peptide pI markers for capillary isoelectric focusing. AB - Nineteen fluorescent pH standards or pI markers ranging pH 3.64-10.12 were developed for use in capillary isoelectric focusing using laser-induced fluorescence detection. Tetra- to tridecapeptides containing one cysteine residue were designed to focus sharply at their respective isoelectric points by including amino acids that contain charged side chains, the pKa values of which are close to the corresponding pI values. An iodoacetylated derivative of tetramethylrhodamine was coupled to the thiol group of cysteine to yield fluorescent pI markers. The pI values of the labeled peptides were precisely determined after isoelectric focusing on polyacrylamide gel slabs by direct measurement of the pH of the focused bands. The markers were subjected to capillary isoelectric focusing for 10-15 min in coated capillaries under conditions of low electroosmosis and were detected by means of a scanning laser induced fluorescence detector down to a level of subpicomolar range. The markers permitted the calibration of a wide-range pH gradient formed in a capillary by fluorescence detection for the first time and should facilitate the development of highly sensitive analytical methods based on a combination of capillary isoelectric focusing and laser-induced fluorescence detection. PMID- 11924963 TI - Gas-phase analysis of trimethylamine, propionic and butyric acids, and sulfur compounds using solid-phase microextraction. AB - Complaints due to odors are an important problem for the wastewater, composting, and animal agriculture industries. Accurate, objective measurement techniques are needed to monitor emissions, to develop new waste handling procedures, and to reduce the production of these volatile gases. Solid-phase microextraction was investigated as a technique for the determination of representative odorous gases. A flow-through Teflon chamber was used to expose the fibers to certified gas standards. A 75-microm carboxen-poly(dimethylsiloxane) (Car-PDMS) coating was used for trimethylamine (TMA), carbon disulfide (CS2), dimethylsulfide (DMS), and dimethyl disulfide (DMDS), and an 85-microm polyacrylate coating was used for propionic acid (PA) and butyric acid (BA). Using a 1-h fiber exposure time and a flow rate through the chamber of 72 mL/min, method detection limits were 2.38, 0.074, 0.150, 0.063, 1.85, and 1.32 ppbv for TMA, DMS, CS2, DMDS, PA, and BA, respectively. Enhanced detector signal was observed for all analytes under flow conditions, as compared to static conditions, and the porous nature of the Car PDMS coating appears to increase the time needed for analytes to reach equilibrium under flow conditions. PMID- 11924965 TI - Simple multiplex genotyping by surface-enhanced resonance Raman scattering. AB - The accurate detection of DNA sequences is essential for a variety of post human genome projects including detection of specific gene variants for medical diagnostics and pharmacogenomics. A specific DNA sequence detection assay based on surface-enhanced resonance Raman scattering (SERRS) and an amplification refractory mutation system (ARMS) is reported. Initially, generation of PCR products was achieved by using specifically designed allele-specific SERRS active primers. Detection by SERRS of the PCR products confirmed the presence of the sequence tested for by the allele-specific oligonucleotides. This lead directly to the multiplex genotyping of human DNA samples for the deltaF508 mutational status of the cystic fibrosis transmembrane conductance regulator gene using SERRS active primers in an ARMS assay. Removal of the unincorporated primers allowed fast and accurate analysis of the three genotypes possible in this system in a multiplex format without any separation of amplicons. The results indicate that SERRS can be used in modern genetic analysis and offers an opportunity for the development of novel assays. This is the first demonstration of the use of SERRS in multiplex genotyping and shows potential advantages over fluorescence as a detection technique with considerable promise for future development. PMID- 11924964 TI - Demonstration of four immunoassay formats using the array biosensor. AB - The ability of a fluorescence-based array biosensor to measure and quantify the binding of an antigen to an immobilized antibody has been demonstrated using the four different immunoassay formats: direct, competitive, displacement, and sandwich. A patterned array of antibodies specific for 2,4,6-trinitrotoluene (TNT) immobilized onto the surface of a planar waveguide and used to measure signals from different antigen concentrations simultaneously. For direct, competitive, and displacement assays, which are one-step assays, measurements were obtained in real time. Dose-response curves were calculated for all four assay formats, demonstrating the array biosensor's ability to quantify the amount of antigen present in solution. PMID- 11924966 TI - A flow injection on-line multiplexed sorption preconcentration procedure coupled with flame atomic absorption spectrometry for determination of trace lead in water, tea, and herb medicines. AB - One of the limitations in previous flow injection (FI) sorption preconcentration procedures in a knotted reactor (KR), which have been carried out exclusively with a single continuous sample injection over a certain period, is the relatively low retention efficiency (typically 40-50%). Although the sensitivity of such systems could be improved by properly increasing sample preconcentration time, sample loading flow rate, or both, further improvement of the sensitivity has been limited by the narrow linearity of the relationship between signal intensity and preconcentration time or sample loading time. In this work, a novel on-line FI multiplexed sorption preconcentration procedure with repetitive sample injections was developed to overcome the above problems in the previous systems. In contrast to previous FI preconcentration systems, the proposed multiplexed preconcentration procedure evenly divides a single longer sample injection step into several shorter substeps while the total preconcentration time is still kept constant. To demonstrate its merits, the proposed FI on-line KR multiplexed sorption preconcentration system was combined with flame atomic absorption spectrometry (FAAS) for determination of trace lead in water, tea, and herb medicines. The lead in the sample solution on-line reacted with ammonium pyrrolidine dithiocarbamate, and the resultant analyte complex was sorbed on the inner walls of the KR. The residual sample solution was then removed from the KR with an air flow. The above two steps were repeated eight times with a total preconcentration time of 120 s. The sorbed analyte was eluted from the KR with 4.5 mol L(-1) HCl for on-line FAAS detection. The present multiplexed preconcentration procedure with eight repetitive sample injections for a total preconcentration time of 120 s gave a retention efficiency of 92%, twice that obtained by one single sample injection preconcentration (47%). In addition, the linear ranges of the diagrams of absorbance against sample loading flow rate and sample loading time were extended, offering more potential for achieving high sensitivity by increasing sample loading rates or sample loading time compared to the previous one single continuous sample injection preconcentration procedure. At a sample loading flow rate of 3.6 mL min(-1) for a total preconcentration period of 120 s, an enhancement factor of 57 and a detection limit (3sigma) of 8 microg L(-1) were obtained. The precision was 1.4% (RSD, n = 11) at the 200 microg L(-1) level. The developed method was successfully applied to the determination of trace lead in various water samples, herb medicines, and a certified tea reference material. PMID- 11924967 TI - Direct determination of benzodiazepines in biological fluids by restricted-access solid-phase microextraction. AB - A biocompatible solid-phase microextraction (SPME) fiber was prepared using an alkyl-diol-silica (ADS) restricted-access material as the SPME coating. The ADS SPME fiber was able to simultaneously fractionate the protein component from a biological sample, while directly extracting several benzodiazepines, overcoming the present disadvantages of direct sampling in biological matrixes by SPME. The fiber was interfaced with an HPLC-UV system, and an isocratic mobile phase was used to desorb, separate, and quantify the extracted compounds. The calculated clonazepam, oxazepam, temazepam, nordazepam, and diazepam detection limits were 600, 750, 333, 100, and 46 ng/mL in urine, respectively. The method was confirmed to be linear over the range of 500-50000 ng/mL with an average linear coefficient (R2) value of 0.9918. The injection repeatability and intraassay precision of the method were evaluated over 10 injections, resulting in a RSD of approximately 6%. The ADS-SPME fiber was robust and simple to use, providing many direct extractions and subsequent determination of benzodiazepines in biological fluids. PMID- 11924968 TI - A selective optode membrane for histidine based on fluorescence enhancement of meso-meso-linked porphyrin dimer. AB - A plasticized polymer film, poly(vinyl chloride) (PVC) incorporated with a specific porphyrin dimer, is shown to exhibit significant and analytical usefulness for optical response toward histidine. The porphyrin dimer containing a free-base porphyrin and a covalently linked metalloporphyrin is shown to be weakly fluorescent as a result of the photoinduced intramolecular electron transfer from the inner free-base porphyrin in singlet excited state to a low spin cobalt(II). The fluorescence enhancement of the membrane by histidine is based on favorable extraction of histidine into the bulk organic membrane and complexation with the inner metallopophyrin moiety and inhibiting the electron transfer process. With the optode membrane described, histidine in a sample solution from 0.0045 to 1.53 mM can be determined. The calibration curve of the optode membrane for histidine shows a good correlation with the mathematically derived formalism and, thus, confirms the theoretically expected behavior. The sensor presented exhibits high selectivity toward histidine over several amino acids and common inorganic anions. The optical selectivity coefficients obtained for histidine over other biologically relevant amino acids and anions are shown to meet the selectivity requirements for the monitoring concentration levels of histidine in biological samples. The selective characteristic of the sensor has been discussed in the view of the coordination chemistry of metalloporphyrin. PMID- 11924970 TI - Controls and reliable conclusions. PMID- 11924969 TI - Enhancement of ionization efficiency by electrochemical reaction products in on line electrochemistry/electrospray ionization Fourier transform ion cyclotron resonance mass spectrometry. AB - A miniaturized two-electrode electrochemical (EC) cell was developed and was coupled on-line with an electrospray ionization Fourier transform ion cyclotron resonance mass spectrometer (ESI-FTICR MS). Electrochemistry on-line with mass spectrometry, EC/ESI-FTICR MS, of triphenylamine (TPA), which undergoes one electron oxidation to form a radical cation (TPA*+), demonstrates a significant sensitivity enhancement compared to ESI-FTICR MS. The on-line EC cell configuration with a stainless steel ES needle as the working electrode produces the highest sensitivity in EC/ESI-MS. The results provide evidence that, during the ES ionization, electrolytic reactions occur mainly in the ES tip region, as previously predicted. The results demonstrate that ESI-MS signal suppression by tetrabutylammonium perchlorate electrolyte, which can be a problem, is minimized in EC/ESI-MS. TPA*+ dimer tetraphenylbenzidine (TPB) can be detected by EC/ESI MS, together with TPA*+, as TPB*+ and TPB2+. The high mass resolving power of FTICR MS was exploited to identify TPB2+ dication in the presence of [TPA*+ - H*]+ ions of the same m/z, from their respective isotopic distributions. The dimer dication TPB2+ can be detected only in EC/ESI-MS. PMID- 11924971 TI - Whole-surface analysis of semiconductor wafers by accumulating short-time mapping data of total-reflection X-ray fluorescence spectrometry. AB - Total-reflection X-ray fluorescence (TXRF) spectrometry with no chemical preconcentration, often called "straight-TXRF", is now widely used in the semiconductor industry. The small detection area of TXRF enablesmapping measurement of contamination of the semiconductor surface, which is very useful in process characterization. However, the small detection area had been believed to limit rapid whole-surface analysis. Contrary to this general understanding, in this study we demonstrated that a new method, called "sweeping-TXRF", which is essentially short-time multipoint mapping by straight-TXRF, can rapidly provide an average concentration. A considerable problem of this method is the contribution of errors in glancing angle and areal element distribution to the fluorescence. Using statistics, we examined the errors and demonstrated that most of them are canceled and are not significant in actual semiconductor applications. The results of an experiment that measured localized 6 x 10(10) atoms cm(-2) nickel contamination supported the above conclusion. Applying sweeping-TXRF to existing TXRF instruments is easy-the only requirement is a small software modification. We believe that sweeping-TXRF will be utilized for rapid whole-surface analysis in many fields, especially in the semiconductor industry. PMID- 11924972 TI - Method for measuring oxygen diffusion coefficients of polymer films by luminescence quenching. AB - To ascertain the relationship between the physical properties of polymer supports and the observed response of luminescence-based oxygen sensors, a quenching-based method was developed to measure oxygen diffusion in polymers. The method offers advantages over existing quenching-based techniques since it allows a simple correction for films of high optical density, and the computations do not assume uniform oxygen concentration throughout the film. Diffusion coefficients (D) were measured for a series of sensors with [Ru(Ph2phen)3]Cl2 (Ph2phen = 4,7-diphenyl 1,10-phenanthroline) asthe luminophore and polystyrene, poly(trimethylsilylmethyl methacrylate), poly(butyl methacrylate), poly(trimethylsilylmethyl methacrylate co-butyl methacrylate), or poly(trimethylsilylmethyl methacrylate-co-1H, 1H heptafluorobutyl methacrylate) as the support. The solvent from which the films were cast was varied, and filler materials such as hydrophobic, amorphous silica or tributyl phosphate plasticizer were added. Results are interpreted by a domain model in which the local environment of the sensor, rather than the bulk properties of the polymer, is the most critical parameter in sensor design. PMID- 11924973 TI - Performances and limits of a parallel oscillator for electrochemical quartz crystal microbalances. AB - This paper describes a driving circuit for an electrochemical quartz crystal microbalance (EQCM) adapted to a wide range of applications. The oscillator is a Miller-type parallel oscillator using an operational transconductance amplifier (OTA). A theoretical study of the oscillating circuit led to the analytical expression of the microbalance frequency as well as to an overestimation of the error on the mass measurement. The reliability of the EQCM was then experimentally verified through electrochemical copper deposition and dissolution. The limit of operation of the EQCM was also investigated, both analytically and experimentally. This work shows that parallel oscillators using few electronic components allow a very reliable EQCM to be obtained for mass measurements on metallic films, even if they are highly damped. PMID- 11924974 TI - Quantitative trace element analysis of individual fly ash particles by means of X ray microfluorescence. AB - A new quantification procedure was developed for the evaluation of X-ray microfluorescence (XRF) data sets obtained from individual particles, based on iterative Monte Carlo (MC) simulation. Combined with the high sensitivity of synchrotron radiation-induced XRF spectroscopy, the method was used to obtain quantitative information down to trace-level concentrations from micrometer-sized particulate matter. The detailed XRF simulation model was validated by comparison of calculated and experimental XRF spectra obtained for glass microsphere standards, resulting in uncertainties in the range of 3-10% for the calculated elemental sensitivities. The simulation model was applied for the quantitative analysis of X-ray tube and synchrotron radiation-induced scanning micro-XRF spectra of individual coal and wood fly ash particles originating from different Hungarian power plants. By measuring the same particles by both methods the major, minor, and trace element compositions of the particles were determined. The uncertainty of the MC based quantitative analysis scheme is estimated to be in the range of 5-30%. PMID- 11924975 TI - In-channel electrochemical detection for microchip capillary electrophoresis using an electrically isolated potentiostat. AB - A new electrode configuration for microchip capillary electrophoresis (CE) with electrochemical (EC) detection is described. This approach makes it possible to place the working electrode directly in the separation channel. The "in-channel" EC detection was accomplished without the use of a decoupler through the utilization of a specially designed, electrically isolated potentiostat. The effect of the working electrode position on the separation performance (in terms of plate height and peak skew) of poly(dimethylsiloxane)-based microchip CEEC devices was evaluated by comparing the more commonly used end-channel configuration with this new in-channel approach. Using catechol as the test analyte, it was found that in-channel EC detection decreased the total plate height by a factor of 4.6 and lowered the peak skew by a factor of 1.3. A similar trend was observed for the small, inorganic ion nitrite. Furthermore, a fluorescent and electrochemically active amino acid derivative was used to directly compare the separation performance of in-channel EC detection to that of a widely used laser-induced fluorescence (LIF) detection scheme. In this case, it was found that the plate height and peak skew for both detection schemes were essentially equal, and the separation performance of in-channel EC detection is comparable to LIF detection. PMID- 11924976 TI - Improved sensitivity of a histamine sensor using an engineered methylamine dehydrogenase. AB - Methylamine dehydrogenase (MADH) may be immobilized in a polypyrrole (PPy) film on an electrode surface and used as an amperometric sensor for the determination of histamine. Using site-directed mutagenesis, phenylalanine 55 on the alpha subunit of MADH was converted to alanine. This alphaF55A MADH exhibits a 400-fold lower Km value for histamine than does native MADH when assayed in solution. An alphaF55A MADH-PPy sensor was constructed, and its properties were compared to that of the native MADH-PPy sensor. The alphaF55A MADH immobilized on the electrode exhibited Michaelis-Menten behavior in response to varied concentrations of histamine with an approximately 3-fold lower Km value than that exhibited by the immobilized native MADH. The detection limit for the native MADH PPy sensor was approximately 20 microM while the alphaF55A MADH-PPy sensor exhibited a detection limit of approximately 5 microM, a 4-fold increase compared to the native MADH-PPy sensor. This work highlights the potential value of using site-directed mutagenesis to engineer enzymes to alter and improve biosensor performance. PMID- 11924977 TI - Biomolecular recognition on well-characterized beads packed in microfluidic channels. AB - We describe a new approach for the analysis of biomolecular recognition in microfluidic channels. The method involves real-time detection of soluble molecules binding to receptor-bearing microspheres, sequestered in affinity column format inside a microfluidic channel. Identification and quantitation of analytes occurs via direct fluorescence measurements or fluorescence resonance energy transfer (FRET). We establish a model system that detects the FLAG epitope. The assay can potentially detect subfemtomole quantities of antibody with a high signal-to-noise ratio and a large dynamic range spanning nearly 4 orders of magnitude in analyte concentration in microliter-to-submicroliter volumes of analyte fluid. Kinetic and equilibrium constants for the reaction of this receptor-ligand pair are obtained through modeling of kinetic responses of the affinity microcolumn and are consistent with those obtained by flow cytometry. Because of the correlation between kinetic and equilibrium data obtained for the microcolumns, quantitative analysis can be done prior to the steady-state end point of the recognition reaction. This method has the promise of combining the utility of affinity chromatography with the advantage of direct, quantitative, and real-time analysis and the cost-effectiveness of microanalytical devices. The approach has the potential to be generalized to a host of bioaffinity assay methods including analysis of protein complexes and molecular assembly and microsystem-based multianalyte determinations. PMID- 11924978 TI - In situ FT-IR measurements of competitive vapor adsorption into porous thin films containing silica nanoparticles. AB - Vapor adsorption into porous ultrathin films on a gold surface is investigated with in situ surface plasmon resonance (SPR) and polarization-modulation infrared reflection-absorption spectroscopy (PM-IRRAS). The thin films are prepared by the electrostatic self-assembly of oppositely charged poly(L-lysine) (PL) and silica nanoparticles on a chemically modified gold surface. Characterization with ex situ SPR and PM-IRRAS demonstrates the buildup of multiple PL/SiO2 bilayers as well as an excellent correlation between the quantitative results from these two techniques. In situ vapor adsorption experiments with these thin films show evidence of porosity, reproducibility, and rapid reversibility. Exposure to acetone vapor (P/P0 = 0.032) causes the film to adsorb 9% acetone by volume, which corresponds to coverage of approximately one-half of the silica nanoparticle surface area. In situ PM-IRRAS provides much information about the molecular interactions occurring in the film upon adsorption or desorption of vapors. Dosing with a mixture of vapors leads to a competition for adsorption into the film, and PM-IRRAS results show that acetone slightly outcompetes nitromethane. These experiments with nanoparticle thin films demonstrate the advantages of using in situ PM-IRRAS for studying reversible adsorption in the presence of vapor mixtures. PMID- 11924979 TI - Development of microwave-assisted drying methods for sample preparation for dried spot micro-X-ray fluorescence analysis. AB - Although dried spot micro X-ray fluorescence (MXRF) is an effective analytical technique for trace elemental analysis, the sample preparation procedures currently used suffer from a number of drawbacks. These drawbacks include relatively long preparation times, lack of control of the sample preparation environment, and possibility of loss of volatile analytes during the drying process. Microwave-assisted drying offers several advantages for dried spot preparation, including control of the environment and minimized volatility because of the differences between microwave heating and conventional heating. A microwave-assisted drying technique has been evaluated for use in preparing dried spots for trace analysis. Two apparatus designs for microwave drying were constructed and tested using multielement standard solutions, a standard reference material, and a "real-world" semiconductor cleaning solution. Following microwave-assisted drying of these aqueous samples, the residues were redissolved and analyzed by ICPMS. Effective recovery was obtained using the microwave drying methods, demonstrating that the microwave drying apparatus and methods described here may be more efficient alternatives for dried spot sample preparation. PMID- 11924980 TI - Two-beam fluorescence cross-correlation spectroscopy in an electrophoretic mobility shift assay. AB - Two-beam fluorescence cross-correlation spectroscopy (FCCS) was used to resolve the bound and unbound fractions of fluorescently labeled single-stranded DNA (ssDNA) in a ssDNA-protein complex as the analyte solution flowed continuously through an electrophoresis capillary. Cross-correlation of the single molecule fluorescence from two spatially separate excitation laser beams resulted in cross correlation functions that consisted of well-resolved peaks characteristic of the different electrophoretic flow velocities of the bound and unbound ssDNA. This decoupled the molecular parameters of the bound and unbound ssDNA used to model the cross-correlation function, which enabled the relative concentrations to be determined without prior knowledge of the pure-component cross-correlation functions, as would be required in an analogous autocorrelation analysis. The relative concentrations of the bound and unbound ssDNA were determined by two beam FCCS within 2-6% precision, even for samples that contained as little as 5% unbound ssDNA, and were consistent with the results obtained by capillary electrophoresis (CE) separation of the same samples. Data sufficient to obtain these results was acquired in 10-15 s per sample. Fluorescently labeled poly(dT)39 complexed with the single stranded DNA binding protein of Escherichia coli served as the model system. The measured dissociation constant of 2.5+/-0.9 nM agreed with the literature value for this complex within experimental error. The CE/two-beam FCCS experiment described here is part of a family of techniques that use single molecule fluorescence detection to resolve different components in an electrophoresis system. Advantages of these methods relative to separations based CE include enhanced sensitivity, the potential for higher speed analyses, elimination of the sample plug injection step, and the ability to carry out the analysis in shorter flow channels. PMID- 11924981 TI - Complete electrolysis using a microflow cell with an oil/water interface. AB - A novel microflow cell with a nitrobenzene (NB)/water (W) interface was developed. A poly(tetrafluoroethylene) membrane filter was employed to prepare the NB/W interface which was formed over a thin channel (0.1 mm thick, 48 cm long) on a silver plate. The silver plate was electrolyzed in advance and served as an Ag/AgCl electrode for controlling the Galvani potential difference across the NB/W interface as well as detecting the current flowing through the interface. Using the microflow cell, complete electrolysis was accomplished for the interfacial transfer of a representative ion (i.e., tetramethylammonium ion). Thus, the present microflow cell was shown to be promising for coulometric (i.e., absolute quantitative) analysis of ions. Also, the microflow cell was shown to be useful for determination of the number of electrons for complicated charge transfer processes at the oil/water interface. PMID- 11924982 TI - Gel protein capillary extraction apparatus. electronic protein transfer. AB - A gel protein capillary extraction apparatus is developed and demonstrated for its rapid and effective transfer of SDS-protein complexes from polyacrylamide gel to a fused-silica capillary. The small dimensions of capillary columns permit the application of high voltages for achieving rapid and effective transfer of gel proteins. Furthermore, the fused-silica capillaries are internally coated with polyacrylamide for the elimination of electroosmotic pumping and protein adsorption onto the capillary wall. The extracted proteins are present in a highly concentrated solution plug as the result of field amplification and sample stacking during the extraction process. Three model proteins, including cytochrome c (14 kDa), ovalbumin (45 kDa), and beta-galactosidase (116 kDa), are visualized using coomassie blue staining and electrophoretically extracted from the gels with protein loading as low as 50 ng. The SDS-cytochrome c complexes extracted from a 50-ng protein loading are concentrated in a 30-nL solution plug inside the capillary with an estimated concentration of 0. 1 mg/mL or 10(-5) M. The capillary format allows the straightforward integration of a miniaturized trypsin-membrane reactor for on-line proteolytic digestion and ESI-MS analysis for protein/peptide identification. PMID- 11924983 TI - Single-channel microchip for fast screening and detailed identification of nitroaromatic explosives or organophosphate nerve agents. AB - A single-channel chip-based analytical microsystem that allows rapid flow injection measurements of the total content of organic explosive or nerve agent compounds, as well as detailed micellar chromatographic identification of the individual ones, is described. The protocol involves repetitive rapid flow injection (screening) assays--to provide a timely warning and alarm--and switching to the separation (fingerprint identification) mode only when harmful compounds are detected. While micellar electrokinetic chromatography, in the presence of sodium dodecyl sulfate (SDS), is used for separating the neutral nitroaromatic explosive and nerve agent compounds, an operation without SDS leads to high-speed measurements of the "total" explosives or nerve agent content. Switching between the "flow injection" and "separation" modes is accomplished by rapidly exchanging the SDS-free and SDS-containing buffers in the separation channel. Amperometric detection was used for monitoring the separation. Key factors influencing the sample throughput, resolution, and sensitivity have been assessed and optimized. Assays rates of about 360 and 30/h can thus be realized for the "total" screening and "individual" measurements, respectively. Ultimately, such development will lead to the creation of a field-deployable microanalyzer and will enable transporting the forensic laboratory to the sample source. PMID- 11924984 TI - Molecularly imprinted polymer coatings for open-tubular capillary electrochromatography prepared by surface initiation. AB - Molecularly imprinted polymer coatings were synthesized in fused-silica capillary columns by the use of a surface-coupled radical initiator. The coatings were prepared using either toluene, dichloromethane, or acetonitrile in the prepolymerization mixtures and were 0.15-2 microm thick as determined by scanning electron microscopy. Solvent-dependent differences in appearance were observed. All the molecularly imprinted polymer-based open-tubular capillary columns were able to separate the enantiomers of propranolol by means of electrochromatography. Electrochromatographic performance was found to be dependent on the type of solvent used during the synthesis. PMID- 11924985 TI - Collection, storage, and filtration of in vivo study samples using 96-well filter plates to facilitate automated sample preparation and LC/MS/MS analysis. AB - The benefits of high-throughput bioanalysis within the pharmaceutical industry are well established. One of the most significant bottlenecks in bioanalysis is transferring in vivo-generated study samples from their collection tubes during sample preparation and extraction. In most cases, the plasma samples must be stored frozen prior to analysis, and the freeze/thaw (F/T) process introduces thrombin clots that are capable of plugging pipets and automated liquid-transfer systems. A new approach to dealing with this problem involves the use of Ansys Captiva 96-well 20-microm polypropylene filter plates to collect, store frozen, and filter plasma samples prior to bioanalysis. The samples are collected from the test subjects, and the corresponding plasma samples are placed directly into the wells of the filter plate. Two Duoseal (patent pending) covers are used to seal the top and bottom of the plate, and the plate is stored at down to -70 degrees C. Prior to sample analysis, the seals are removed and the plate is placed in a 96-well SPE manifold. As the plasma thaws, it passes (by gravity or mild vacuum) through the polypropylene filter into a 96-well collection plate. A multichannel pipet or automated liquid-transfer system is used to transfer sample aliquots without fear of plugging. A significant advantage of this approach is that, unlike other methods, issues related to incomplete pipetting are virtually eliminated. The entire process is rapid since thawing and filtering take place simultaneously, and if a second F/T cycle is required for reanalysis, it is not necessary to refilter the samples (additional clotting was not observed after three F/T cycles). This technique was tested using monkey, rat, and dog plasma and sodium heparin and EDTA anticoagulants. To assess the possibility of nonspecific binding to the polypropylene filter, a variety of drug candidates from diverse drug classes were studied. Validation data generated for two Lilly compounds from distinct classes, before and after filtering, are presented in this paper as practical examples of this technique. While LC/MS/MS is the primary method of bioanalysis in our laboratory, the technique presented in this paper is applicable to other forms of detection as well. PMID- 11924986 TI - Selective detection of o-diphenols on copper-plated screen-printed electrodes. AB - Selective detection of o-diphenols (e.g., catechol, dopamine, and pyrogallol) in the presence of simple phenols, m- and p- derivative diphenols, and ascorbic acid has been demonstrated on copper-plated screen-printed electrodes (CuSPEs) in pH 7.4 phosphate buffer solution. The CuSPE showed an unusual catalytic response at 0.05 V versus Ag/AgCl selectively to o-diphenolic compounds. The o-diphenols can thus be determined amperometrically through direct electrochemical oxidation in low potentials (approximately 0 V), where the CuSPE is much less subject to interfering reactions. Such a catalytic phenomenon cannot be observed on conventional Pt and glassy carbon electrodes. The selective mechanism is explained in terms of the formation of cyclic five-member complex intermediate (Cu(II)-o-quinolate). Most important of all, the common drawbacks of electrode fouling through polymerization were completely overcome in this system. PMID- 11924987 TI - Comments on "Determination of low molecular weight silicones in plasma and blood of women after exposure to silicone breast implants by GC/MS". PMID- 11924988 TI - Antibodies choose the "right" catalysts. PMID- 11924989 TI - The science of detecting terror. PMID- 11924990 TI - False cyanide detection. PMID- 11924992 TI - ACS founders were analytical chemists, too. American Chemical Society. PMID- 11924993 TI - Software for MS protein identification. PMID- 11924991 TI - Isotopic analysis of dinosaur bones. PMID- 11924994 TI - Immunoassay with multicomponent protein microarrays fabricated by electrospray deposition. AB - Two new techniques were recently developed in these laboratories for fabrication of protein microarrays: electrospray deposition of dry proteins and covalent linking of proteins from dry deposits to a dextran-grafted surface. Here we apply these techniques to simultaneously fabricate 1200 identical microarrays. Each microarray, 0.6 x 0.6 mm2 in size, consists of 28 different protein antigens and allergens deposited as spots, 30-40 microm in diameter. The ability of the microarrays to detect IgG antibodies in plasma samples from mice immunized with different sets of antigens and IgE antibodies in human plasmas was examined using ELISA. Comparison of the microarray-based ELISAs with standard ELISAs in microtiter plates established that both techniques provided identical responses in 88% of all the antibody/antigen interactions tested. Both techniques showed similar antibody detecting sensitivity defined by the maximum dilution of serum at which a reliable signal distinguishable from the background was obtained. PMID- 11924995 TI - One-step concentration of analytes based on dynamic change in pH in capillary zone electrophoresis. AB - A novel strategy for one-step concentration of analytes during capillary electrophoresis (CE) is presented. A short platinum wire was inserted into the 75 microm-i.d. separation capillary. When a high voltage was applied for CE separation, a sharp pH gradient along the capillary was created dynamically by the electrolysis of water in the running buffer. Concentration of a large volume of injected analytes was accomplished by the change in analyte charge due to the dynamic pH gradient. Depending on the polarity of the applied potential and the direction of electroosmotic flow, either anions or cations can be concentrated. Several hundredfold concentration factors were achieved. Fluorescence imaging by a CCD camera was used to monitor 10 cm of the capillary near the platinum wire during the concentration process. The observations are consistent with a sweeping mechanism. PMID- 11924996 TI - Quantitative profiling of phospholipids by multiple precursor ion scanning on a hybrid quadrupole time-of-flight mass spectrometer. AB - A hybrid quadrupole time-of-flight mass spectrometer featured with ion trapping capabilities was employed for quantitative profiling of total extracts of endogenous phospholipids. Simultaneous acquisition of precursor ion spectra of multiple fragment ions allowed detection of major classes of phospholipids in a single experiment. Relative changes in their concentration were monitored using a mixture of isotopically labeled endogenous lipids as a comprehensive internal standard. Precursor ion scanning spectra were acquired simultaneously for acyl anions of major fatty acids in negative ion mode and identified the fatty acid moieties and their relative position at the glycerol backbone in individual lipid species. Taken together, a combination of multiple precursor ion scans allowed quantitative monitoring of major perturbation in phospholipid composition and elucidating of molecular heterogeneity of individual lipid species. PMID- 11924998 TI - Scintillation proximity assay using molecularly imprinted microspheres. AB - Molecularly imprinted microspheres were prepared as antibody binding mimics and used in scintillation proximity assay of a beta-adrenergic antagonist, (S) propranolol. By using small polymer beads, we were able to place an organic scintillator and an "antenna" component in close proximity to the imprinted binding sites. When the radioactive template bound to the polymer, radiation energy was effectively transferred, via the antenna component, to the scintillator to generate a fluorescence signal. Using molecularly imprinted microspheres instead of antibodies, we have demonstrated competitive scintillation proximity assays for (S)-propranolol in both organic and aqueous solvents. The experimental results were further validated by normal ligand binding analysis, where liquid scintillation counting was used for quantification. PMID- 11924997 TI - Quantifying peptides in isotopically labeled protease digests by ion mobility/time-of-flight mass spectrometry. AB - Ion mobility/time-of-flight techniques have been used to analyze mixtures of isotopically labeled peptides. The isotopic labels were generated by treatment of peptides with N-acetoxysuccinimide (or the deuterated analogue), which results in acetylation (or deuterioacetylation) of the primary amines (i.e., the N-terminus and lysine residues). The relative concentrations of a peptide in each sample are determined by comparing the peak intensities for isotopic pairs. An important consideration is that as mixtures become increasingly complex, isotopic pairs of peaks may overlap with other peaks in the mass spectrum. The influence of the acetyl and deuterioacetyl groups on the mobilities of peptides is considered. The coincidence in mobilities of isotopic pairs provides a means of distinguishing isotopic pairs from other isobaric interferences. PMID- 11924999 TI - Identification of archaeological adhesives using direct inlet electron ionization mass spectrometry. AB - Adhesives made from natural substances such as resins, tars, and waxes are found during excavations on archaeological sites dating back to prehistoric periods. Until now, their analysis was mainly performed by gas chromatography, possibly coupled to mass spectrometry, after extraction, purification, and derivatization of the samples. To minimize sampling and sample preparation of ancient organic remains, which are often preserved in tiny amounts, we have directly analyzed archaeological samples from Bronze and Iron Age periods by direct inlet electron ionization mass spectrometry. A series of contemporary natural and synthetic substances, including pine and pistacia resins, birch bark tar, beeswax, and plant oils, possibly used for adhesive fabrication during ancient times, was also investigated with the same technique as reference materials. Despite the complexity of their chemical composition, pine resin and birch bark tar were clearly identified in archaeological samples. Furthermore, mass spectrometry has been shown to be efficient for the identification of glues made of a mixture of beeswax, presenting a series of mass spectral peaks assigned to long-chain esters, and birch bark tar, whose mass spectrum presents characteristic peaks of lupane compounds. The intentional mixing of birch bark tar and beeswax during prehistory is reported here for the first time. PMID- 11925001 TI - Stabilization of anionic adducts in negative ion electrospray mass spectrometry. AB - Attachment of small anions to neutral molecules is an important ionization mechanism in negative ion electrospray mass spectrometry. In this report, the tendency for different anions to remain attached to selected analyte compound classes has been systematically investigated. A rationale for the formation and stability of preferred anionic adducts is proposed in light of thermodynamic considerations. A series of collision-induced dissociation experiments reveals that the gas-phase basicities of the deprotonated analyte molecule ([M - H]-) and the anion moiety play important roles in determining the stability of anionic adducts. Adducts of the form [M - H]-...H+...[anion]- manifest increased stability when the two anions have similar gas-phase basicities. Within certain limitations, the difference in deltaG degrees values for proton combination with [M - H]- and with [anion]- can be used as a first-order predictor of adduct stability. In addition, stability increases with the rising gas-phase basicities of the two moieties. The specific interaction between a small inorganic anion (bisulfate) and a neutral analyte molecule (alpha-D-glucose) in the form of multiple hydrogen bonding has also been affirmed by computer modeling to contribute to the stability of some anionic adducts. Last, the gas-phase basicity of deprotonated alpha-D-glucose (i.e., the gas-phase acidity of alpha-D-glucose) is determined by a "bracketing method" to be in the range of 1373-1407 kJ/mol. PMID- 11925000 TI - Oligonucleotide mixture analysis via electrospray and ion/ion reactions in a quadrupole ion trap. AB - Electrospray ionization combined with ion/ion reactions in a quadrupole ion trap can be used for the direct analysis of oligonucleotide mixtures. Elements to the success of this approach include factors related to ionization, ion/ion reactions, and mass analysis. This paper deals with issues regarding the ion polarity combination, viz., positive oligonucleotides/negative charge-transfer agent versus negative oligonucleotides/positive charge-transfer agent. Anions derived from perfluorocarbons appear to be directly applicable to mixtures of positive ions derived from electrospray of oligonucleotides, in direct analogy with positive protein ions. Conditions for forming positive oligonucleotide ions devoid of adducts were more difficult to establish than for forming relatively clean negative oligonucleotide ions. A new approach for manipulating negative ion charge states in the ion trap is described and is based on use of the electric field of the positive charge-transfer agent for storage of high-mass negative ions formed during the ion/ion reaction period. Oxygen cations are shown to be acceptable for charge-state manipulation of mixed-base oligomers but induce fragmentation in polyadenylate homopolymers. Protonated isobutylene (C4H9+), on the other hand, is shown to induce significantly less fragmentation of polyadenylate homopolymers. PMID- 11925002 TI - Coupling ion mobility separations, collisional activation techniques, and multiple stages of MS for analysis of complex peptide mixtures. AB - An ion trap/ion mobility/quadrupole/collision cell/time-of-flight mass spectrometer that incorporates a differentially pumped orifice-skimmer cone region at the back of the drift tube has been developed for the analysis of peptide mixtures. The combined approach allows a variety of strategies to be employed for collisionally activating ions, and fragments can be monitored by subsequent stages of mass spectrometry in a parallel fashion, as described previously (Anal. Chem. 2000, 72, 2737). Here, we describe the overall experimental approach in detail. Applications involving different aspects of the initial mobility separation and various collisional activation and parallel sequencing strategies are illustrated by examining several simple peptide mixtures and a mixture of tryptic peptides from beta-casein. Detection limits associated with various experimental configurations and the utility for analysis of complex systems are discussed. PMID- 11925003 TI - Rabbit cone bipolar cells: correlation of their morphologies with whole-cell recordings. AB - The superfused retinal slice preparation was used to examine the morphology and glutamate-activated whole-cell currents of rabbit bipolar cells. There were six morphologically distinct types of cone bipolar cells and a rod bipolar cell that had axon terminals stratifying in stratum 3 to 5 of sublamina-b. All of these bipolar cell types exhibited an outward current in response to the application of the metabotropic glutamate receptor, mGluR6, agonist AP-4 (APB), and had I/V curves indicative of membrane channel closure. Conversely, there were no currents activated during the application of kainate, the AMPA/kainate receptor agonist. These data demonstrate they were on-bipolar cells. In addition, there were six morphologically distinct cone bipolar cells that stratified in sublamina-a. Every cell with axonal arborizations in stratum 1 and 2 exhibited an inward current when the ionotropic glutamate receptor agonist kainate was applied. This current was blocked by application of the AMPA/kainate receptor antagonist CNQX. These cells also decreased their membrane resistance in response to kainate, a characteristic of the opening of channels within the plasma membrane. Without exception, no cells stratifying in sublamina-a responded to the mGluR6 agonist AP 4, further identifying them as off-bipolar cells. PMID- 11925004 TI - Lateral spread of adaptation as measured with the multifocal electroretinogram. AB - We examined whether lateral spread of adaptation can be observed in the electroretinogram in humans. Specifically, we tested whether the luminance level of a surrounding, nonmodulated annulus affects the multifocal electroretinogram (ERG) response of a modulated central area. Multifocal electroretinograms were recorded in response to an array of 37 unscaled hexagons subtending a retinal area of 38 deg x 35 deg. Responses were recorded in six control subjects. In the first series of experiments, only the center hexagon was modulated, while the surrounding 36 hexagons were held constant at either 0.45, 172, or 340 cd/m2. In a subsequent series of control experiments, modulation depth of the center hexagon was varied and the proximity of the surrounding hexagon systematically altered. For the center-modulated condition, response amplitude and implicit time for the first-order kernel response significantly decreased as a function of increasing surround luminance. Control experiments demonstrated that the effect of the surround illumination was not due to scattered light but was influenced by the proximity of the surrounding annulus. These results demonstrate that lateral adaptation influences can be measured using the multifocal ERG. PMID- 11925005 TI - Multiple cell targets for melatonin action in Xenopus laevis retina: distribution of melatonin receptor immunoreactivity. AB - In the retina of the African clawed frog (Xenopus laevis), melatonin is synthesized by the photoreceptors at night, and binds to receptors that likely mediate paracrine responses. Melatonin appears to alter the sensitivity of the retinal cells to light, and may play a key role in regulating important circadian events that occur in the eye. A polyclonal antibody was raised against a 13 amino acid peptide corresponding to a region of the third cytoplasmic loop of the Xenopus laevis Mel1c melatonin receptor. Western blot analysis revealed a major immunoreactive band of approximately 60 kD in neural retina and retinal pigment epithelium (RPE) membranes. Immunocytochemical labeling of sections of Xenopus eyes demonstrated intense melatonin receptor-like immunoreactivity in the inner plexiform layer (IPL). Immunolabeling with antibodies to glutamate decarboxylase (GAD) or tyrosine hydroxylase (TOH) appeared to co-localize with the melatonin receptor immunoreactivity in different sublaminas of the IPL. This suggests that both GABAergic and dopaminergic amacrine cells express melatonin receptor protein. There were also some melatonin receptor immunoreactive varicose fibers in the IPL that did not co-localize with either TOH or GAD, and may represent efferent fibers, since they could be followed into the optic nerve. Melatonin receptor immunoreactivity was also present on cell soma in the ganglion cell layer. Furthermore, a moderate level of melatonin receptor immunoreactivity was observed in the RPE and rod and cone photoreceptor cells. The presence of melatonin receptor immunoreactivity in these cells supports previous observations of melatonin receptor RNA expression in multiple cell types in the Xenopus retina. Expression of melatonin receptor protein in the photoreceptors suggests that melatonin may have a direct action on these cells. PMID- 11925006 TI - AMPA-preferring receptors mediate excitatory non-NMDA responses of primate retinal ganglion cells. AB - Glutamate and kainate-induced currents of primate ganglion cells were studied using the whole-cell patch-clamp technique in a retinal slice preparation. Antagonists and allosteric modulators of desensitization selective for either alpha-amino-3-hydroxy-5-methyl-4-isoazoleprionic acid (AMPA)- or kainate preferring receptors were used to determine the contributions of each type of receptor to excitatory responses. With synaptic transmission and NMDA receptors blocked, the AMPA-preferring receptor antagonist GYKI 52466 (30 microM-100 microM) reversibly blocked most of the glutamate-induced current in ganglion cells. GYKI 52466 also blocked the response in ganglion cells to focally applied kainate, suggesting that the current response to kainate arises from activation of AMPA-preferring receptors, and not kainate-preferring receptors. Both cyclothiazide (10 microM-100 microM) and the novel drug 4-[2 (phenylsulfonylamino)ethylthio]-2,6-difluoro-phenoxyacetamide (PEPA, 20 microM 100 microM), which selectively enhance responses mediated by AMPA-preferring receptors, enhanced glutamate-induced responses of ganglion cells. Since these drugs preferentially inhibit desensitization of the flip and flop splice variants, respectively, of AMPA-preferring receptors, it is likely that both splice variants are present on these ganglion cells. Concanavalin A, which selectively suppresses the desensitization of kainate-preferring receptors, had no effect on the glutamate-induced responses of ganglion cells. We conclude that the non-NMDA component of the excitatory, glutamatergic input to primate ganglion cells is mediated largely by AMPA-preferring receptors, with little, if any, kainate-preferring receptor-mediated response. PMID- 11925008 TI - Cortical and subcortical afferents to the nucleus reticularis tegmenti pontis and basal pontine nuclei in the macaque monkey. AB - Anatomical findings are presented that identify cortical and subcortical sources of afferents to the nucleus reticularis tegmenti pontis (NRTP) and basal pontine nuclei. Projections from the middle temporal visual area (MT), medial superior temporal visual area (MST), lateral intraparietal area (LIP), and areas 7a and 7b to the basal pontine nuclei were studied using 3H-leucine autoradiography. The results complemented a parallel study of retrograde neuronal labeling attributable to injecting WGA-HRP into NRTP and neighboring pontine nuclei. Small 3H-leucine injections confined to MT, MST, LIP, area 7a, or area 7b, produced multiple patches of pontine terminal label distributed as follows: (1) An injection within MT produced terminal label limited to the dorsolateral and lateral pontine nuclei. (2) Injections restricted to MST or LIP showed patches of terminal label in the dorsal, dorsolateral, lateral, and peduncular pontine nuclei. (3) Area 7a targets the dorsal, dorsolateral, lateral, peduncular, and ventral pontine nuclei, whereas area 7b projects, additionally, to the dorsomedial and paramedian pontine nuclei. Notably, no projections were seen to NRTP from any of these cortical areas. In contrast, injections made by other investigators into cortical areas anterior to the central sulcus revealed cerebrocortical afferents to NRTP, in addition to nuclei of the basal pontine gray. With our pontine WGA-HRP injections, retrograde neuronal labeling was observed over a large extent of the frontal cortex continuing onto the medial surface which included the lining of the cingulate sulcus and cingulate gyrus. Significant subcortical sources for afferents to the NRTP and basal pontine nuclei were the zona incerta, ventral mesencephalic tegmentum, dorsomedial hypothalamic area, rostral interstitial nucleus of the medial longitudinal fasciculus, red nucleus, and subthalamic nucleus. The combined anterograde and retrograde labeling data indicated that visuo-motor cortico-pontine pathways arising from parietal cortices target only the basal pontine gray, whereas the NRTP, together with select pontine nuclei, is a recipient of afferents from frontal cortical areas. The present findings implicate the existence of parallel direct and indirect cortico-pontine pathways from frontal motor-related cortices to NRTP and neighboring pontine nuclei. PMID- 11925007 TI - Retinal development of West Australian dhufish, Glaucosoma hebraicum. AB - An investigation of retinal specializations was carried out in larval and juvenile dhufish, Glaucosoma hebraicum (Glaucosomidae, Teleostei). The development of photoreceptors and formation of the retinal mosaic was followed by light and electron microscopy. At hatching the eye was undifferentiated. Cone photoreceptors were present by day 3 posthatch (dph), when exogenous feeding began. Single and multiple cones were present in a row arrangement from 3 dph to 20 dph, when the first rod nuclei were observed. Between 20 dph and approximately 3 months posthatch (mph), the row arrangement was replaced by a square mosaic of four double cones surrounding a single cone, and the cones increased in size, with the outer segments reaching up to 30 microm in length. During the period of spatial rearrangement, triple cones were often observed. From their first appearance, rod photoreceptors were added rapidly. Investigation of ganglion cell topography in 3-mph fish that had attained the adult-like square photoreceptor mosaic was carried out using retinal wholemounts. The highest densities of neurones in the ganglion cell layer were in temporal retina but no well-defined area centralis was observed. Microspectrophotometric measurements of the visual pigments within the outer segments of the photoreceptors of 3-mph fish revealed double cones with identical absorption spectra in each member of the outer segment, and the wavelength of maximum absorption (lambda(max)) located at 522 nm. Single cones were found to possess a visual pigment with lambda(max) at 460 nm and rods with a lambda(max) of 498 nm. The results imply that the larvae and juveniles are adapted for survival in coastal waters and may be active in relatively low light levels from early stages of development. PMID- 11925010 TI - Photoreceptors and visual pigments in the red-eared turtle, Trachemys scripta elegans. AB - Absorbance spectra of cone outer segments and oil droplets were recorded microspectrophotometrically in the retina of the red-eared turtle, Trachemys scripta elegans. There are four cone visual pigments, with lambda(max) = 617 nm (red sensitive), 515 nm (green sensitive), 458 nm (blue sensitive), and 372 nm (UV-sensitive). The red-sensitive pigment resides in single cones with red or orange oil droplets, and in both members of double cones. The principal member of the double cone contains an orange oil droplet, and the accessory member is droplet free. The green-sensitive pigment is situated in single cones with orange/dark yellow droplets. The blue-sensitive pigment is combined with the UV absorbing oil droplet in single cones. The UV-sensitive pigment resides in single cones with clear oil droplets that exhibited virtually no absorbance down to 325 nm. Thus, seven types of cones can be identified based on their morphology, oil droplet color, and the visual pigment absorbance. At the moment, this is the most complex cone system described for vertebrates. PMID- 11925009 TI - Disruption of transient photoreceptor targeting within the inner plexiform layer following early ablation of cholinergic amacrine cells in the ferret. AB - Photoreceptors in the ferret's retina have been shown to project transiently to the inner plexiform layer (IPL) prior to their differentiation of an outer segment. On postnatal day 15 (P-15), when this projection achieves maximal density, the photoreceptors projecting into the IPL extend primarily to one of two depths, coincident with the processes of cholinergic amacrine cells. The present study has used an excitotoxic approach employing subcutaneous injections of L-glutamate to ablate these cholinergic amacrine cells on P-7, in order to see whether their elimination alters this targeting of photoreceptor terminals within the IPL. The near-complete elimination of cholinergic amacrine cells at P-15 was confirmed, although the population of retinal ganglion cells was also affected, being depleted by roughly 50%. The rod opsin-immunopositive terminals in such treated ferrets no longer showed a stratified distribution, being found throughout the depth of the IPL, as well as extending into the ganglion cell layer. This effect should not be due to the partial loss of retinal ganglion cells, however, since optic nerve transection at P-2, which eliminates the ganglion cells entirely while leaving the cholinergic amacrine cell population intact, was shown not to affect the stratification pattern of the photoreceptors within the IPL. These results strongly suggest that the targeting of the photoreceptor terminals to discrete strata within the IPL is dependent upon the cholinergic amacrine cell processes. PMID- 11925011 TI - Tetrachromatic input to turtle horizontal cells. AB - Recent physiological experiments support behavioral and morphological evidence for a fourth type of cone in the turtle retina, maximally sensitive in the ultraviolet (UV). This cone type has not yet been included in the models proposed for connectivity between cones and horizontal cells. In this study, we examined the inputs of UV, S, M, and L cones to horizontal cells. We used the high resolution Dynamic Constant Response Method to measure the spectral sensitivity of horizontal cells without background light and after adaptation to UV, blue (B), green (G), and red (R) light. We concluded the following: (1) Tetrachromatic input to a Y/B horizontal cell was identified. The spectral-sensitivity curves of the cell in three of the adaptation conditions were well represented by L-, M-, and S-cone functions. Adaptation to blue light revealed a peak at 372 nm, the same wavelength location as that determined behaviorally in the turtle. A porphyropsin template could be closely fitted to the sensitivity band in that region, strong evidence for input from a UV cone. (2) The spectral-sensitivity functions of R/G horizontal cells were well represented by the L- and M-cone functions. There was no indication of UV- or S-cone inputs into these cells. (3) The spectral sensitivities of the monophasic horizontal cells were dominated by the L cone. However, the shape of the spectral-sensitivity function depended on the background wavelength, indicating secondary M-cone input. Connectivity models of the outer retina that predict input from all cone types are supported by the finding of tetrachromatic input into Y/B horizontal cells. In contrast, we did not find tetrachromatic input to R/G and monophasic horizontal cells. Chromatic adaptation revealed the spectral-sensitivity function of the turtle UV cone peaking at 372 nm. PMID- 11925012 TI - Molecular cloning and characterization of five opsin genes from the marine flatfish Atlantic halibut (Hippoglossus hippoglossus). AB - Most molecular studies on the visual system in fish have been performed on freshwater teleosts such as goldfish and zebrafish where cones and rods appear simultaneously. Many marine fishes have long larval phase in the upper pelagic zone before transformation into a juvenile and a benthic life style. The retina at the larval stages consists of only single cone cells; later during metamorphosis double cones and rods develop. The flatfish Atlantic halibut (Hippoglossus hippoglossus) is a typical example of a marine species with such a two-step retina development. In this study, we have cloned five different opsins from Atlantic halibut larvae and juvenile retinas. Sequence comparisons with other opsins and phylogenetic analysis show that the five genes belong to the opsins of long-wavelength sensitive (L); middle-wavelength sensitive, M(Cone) and M(Rod); and short-wavelength sensitive, S(Blue) and S(Ultraviolet), respectively. In situ hybridization analysis reveals expression in double cone (L and M(Cone)), single cone (S(Blue) and S(Ultraviolet)), and rod (M(Rod)) types of photoreceptor cells in juvenile halibut retina. The visual system in Atlantic halibut seems therefore to have all four types of cone photoreceptors in addition to rod photoreceptors. This work shows for the first time molecular isolation of a complete set of retinal visual pigment genes from a marine teleost and describes the first cloning of an ultraviolet-sensitive opsin type from a marine teleost. PMID- 11925013 TI - Alterations in NMDA receptor expression during retinal degeneration in the RCS rat. AB - To determine how a progressive loss of photoreceptor cells and the concomitant loss of glutamatergic input to second-order neurons can affect inner-retinal signaling, glutamate receptor expression was analyzed in the Royal College of Surgeons (RCS) rat, an animal model of retinitis pigmentosa. Immunohistochemistry was performed on retinal sections of RCS rats and congenic controls between postnatal (P) day 3 and the aged adult (up to P350) using specific antibodies against N-methyl-D-aspartate (NMDA) subunits. All NMDA subunits (NR1, NR2A-2D) were expressed in control and dystrophic retinas at all ages, and distinct patterns of labeling were found in horizontal cells, subpopulations of amacrine cells and ganglion cells, as well as in the outer and inner plexiform layer (IPL). NRI immunoreactivity in the inner plexiform layer of adult control retinas was concentrated in two distinct bands, indicating a synaptic localization of NMDA receptors in the OFF and ON signal pathways. In the RCS retina, these bands of NRI immunoreactivity in the IPL were much weaker in animals older than P40. In parallel, NR2B immunoreactivity in the outer plexiform layer (OPL) of RCS rats was always reduced compared to controls and vanished between P40 and P120. The most striking alteration observed in the degenerating retina, however, was a strong expression of NRI immunoreactivity in Muller cell processes in the inner retina which was not observed in control animals and which was present prior to any visible sign of photoreceptor degeneration. The results suggest functional changes in glutamatergic receptor signaling in the dystrophic retina and a possible involvement of Muller cells in early processes of this disease. PMID- 11925014 TI - Diurnal variation in synaptic ribbon length and visual threshold. AB - Previous work suggests that photoreceptor synaptic ribbon length and absolute dark-adapted threshold may vary during a 24-h diurnal cycle. To test this hypothesis, we examined the length of photoreceptor synaptic ribbons and the dark adapted threshold in black (+/+) and albino (c2J/c2J) C57BL/6J mice at six times over a 24-h period. Testing began 2 h after light onset (ZT 2:00) and continued at successive 4-h intervals (12 h:12 h light:dark). We determined the length of the synaptic ribbons in frozen sections by labeling them with an antibody specific for synaptic ribbons. Synaptic ribbons vary in length at different points in the diurnal cycle in both types of mice, but the synaptic ribbons in black mice are longer than those in albino mice by an average of 0.33 microm. The synaptic ribbons of black mice also have a larger response to changes in the light cycle. Ribbon length in black mice ranges from 1.66 microm to 1.4 microm, whereas ribbon length in albino mice ranges from 1.32 microm to 1.25 microm. The shortest ribbons are evident 6 h after light onset in both types of mice, whereas the longest ribbons appear within 2 h after light onset. These changes in synaptic ribbon length support the idea that photoreceptor synaptic ribbons are dynamic structures whose length changes over a 24-h diurnal cycle. Examining black and albino mice with a water-maze behavioral assay showed that visual thresholds in black and albino mice vary over the 24-h diurnal cycle. The visual thresholds of albino mice are elevated compared with black mice at all times tested. This is consistent with previous findings of visual thresholds in hypopigmented mice. The lowest threshold (greatest sensitivity) is present 2 h after light onset (ZT 2:00) and corresponds to the time when synaptic ribbons are the longest. The highest threshold is observed 6 h after light onset, the time when synaptic ribbons are shortest. These results show that synaptic ribbon length and visual sensitivity vary together in relation to the time. PMID- 11925015 TI - Voltage-gated calcium and sodium currents of starburst amacrine cells in the rabbit retina. AB - The voltage-gated calcium and sodium currents of starburst amacrine cells were examined in slices of the adult rabbit retina. ON-center starburst amacrine cells were targeted for whole-cell recording by prelabeling the retina with the nuclear dye 4'-6-diamidino-2-phenylindole hydrochloride (DAPI). Calcium currents were isolated using an external Ringer that contained tetrodotoxin to block sodium currents and barium to block potassium channels. When starburst amacrine cells were stepped to holding potentials positive to -50 mV, a series of voltage dependent calcium currents were activated. The calcium current peaked at -10 mV. The calcium currents kinetics were mainly sustained in nature, showing only a small amount of slow inactivation. Nickel (100 microM), a T-type channel blocker, had no effect on the calcium current. Application of the L-type channel agonist BAY K8644 (1-2.5 microM) had small variable effects on the calcium current while the L-type channel antagonist nifedipine (10 microM) had no effect. However, addition of a reported N-type calcium channel antagonist, omega-conotoxin G6A (1 microM), blocked a large portion of the calcium current, as did a more nonselective antagonist, omega-conotoxin M7C (200 nM). Agatoxin 4A (500 nM) reduced a smaller sustained calcium current component, implying a P/Q-type calcium channel was present on these neurons. In addition to the calcium currents, a fast voltage-gated sodium current was observed in many starburst cells. This current could be blocked by tetrodotoxin (200-500 nM). The differing kinetics and durations of the sodium and calcium currents could play important roles in the regulation of synaptic release and in the coordination of spiking by starburst amacrine cell dendrites during retinal development and in the encoding of motion across the retinal surface. PMID- 11925016 TI - Layer differences in the effect of monocular vision in light- and dark-reared kittens. AB - We compared the effect of 2 days of monocular vision on the ocular dominance of cells in the visual cortex of light-reared kittens with the effect in dark-reared kittens at 6, 9, and 14 weeks of age, and analyzed the results by layer. The size of the ocular-dominance shift declined with age in all layers in light-reared animals. There was not a large change in the ocular-dominance shift with age in dark-reared animals in any layer, suggesting that dark rearing largely keeps the cortex in the immature 6-week state until 14 weeks or longer, although there was a slight decrease in layers II, III, and IV, and a slight increase in layers V and VI. At 14 weeks, the difference between light- and dark-reared animals was smallest in layer IV, larger in layers II/III, and largest in layers V/VI, suggesting that dark rearing has a large effect on intracortical synapses and a small effect on geniculocortical synapses. There was a significant ocular dominance shift in layer IV at 14 weeks of age in both light- animals and dark reared animals, showing that the critical period for ocular-dominance plasticity is not ended at this age. While the ocular-dominance shift after 26 h of monocular deprivation in 6-week animals was similar in light- and dark-reared animals, after 14 h it was smaller in dark-reared animals, showing that ocular dominance changes occur more slowly in dark-reared animals at this age, in agreement with Mower (1991). Increases in selectivity for axis of movement after 26 h of monocular vision were seen in dark-reared animals at 6 weeks of age, but not at 9 or 14 weeks of age, showing that the critical period for axial selectivity ends earlier than the critical period for ocular dominance in dark reared animals, as it does in light-reared animals. PMID- 11925018 TI - Asymmetrical dynamics of voltage spread in retinal horizontal cell networks. AB - Lateral voltage spread in electrically coupled retinal horizontal cell networks is the substrate of center-surround antagonism in bipolar and ganglion cells. We studied its spatial and temporal properties in more detail in turtle L1 horizontal cells by using a contrast border as light stimulus. Experimental data were contrasted with expectations from a linear continuum model to specify the impact of nonlinearities. The assumptions for the diffusion term of the continuum model were justified by neurobiotin labeling. Measured voltage spread revealed two different length constants lambda+ and lambda0, under illuminated and nonilluminated regions of the retina, respectively, as predicted by the linear model. Length constants in the illuminated region showed strong temporal dynamics. For the initial phase of the horizontal cell responses lambda+ was larger than lambda0. This was also in accordance with the model. Right at the peak of the response, however, lambda+ dropped below lambda0 and did not change any more. It is this temporal reversal of asymmetry in voltage spread and not the decrease of lambda+ itself that is lacked by the linear model. The observed independence of the mean ratio lambda+/lambda0 from light intensity in both the peak and the plateau phases of horizontal cell responses contradicts the linear assumption, too. These two effects have to be addressed to local nonlinearities in the horizontal cell network like a negative feedback loop from photoreceptors and/or voltage-dependent conductances. Due to the failure of the linear model, firm conclusions about the membrane resistance and the coupling resistance of the horizontal cell network cannot be drawn from length constant measurements. PMID- 11925017 TI - Effects of monocular deprivation and reverse suture on orientation maps can be explained by activity-instructed development of geniculocortical connections. AB - Mature visual cortex shows a single, binocularly matched orientation map. This matching develops without visual experience. It persists despite early monocular deprivation that largely eliminates one eye's map, followed by reverse suture (deprivation of the previously open eye and opening of the previously deprived eye), even though the two eyes lack common visual experience in this case. These results have been interpreted to suggest that the structure of orientation maps either is innately predetermined or, if it arises through self-organization, is determined by external cues such as boundary conditions or a "scaffolding" of horizontal connections. We show, to the contrary, that these results are the expected outcomes if orientation maps develop through activity-instructed, correlation-based development of the geniculocortical connections without additional cues. A weak, binocularly correlated orientation map is known to exist before deprivation onset; we previously showed how this can arise through activity-instructed development. Now we show that this initial correlation between the two eyes' maps can persist or increase despite deprivation sufficient to cause massive loss of the deprived eye's geniculocortical synaptic strength, followed by reverse suture. Given sufficient early correlated map development, each map's fate is "dynamically committed": the two eyes' maps will converge upon a common outcome, even if developing independently. This dynamic fate commitment is retained even after severe deprivation. PMID- 11925019 TI - Progress toward tuberculosis control--India, 2001. AB - Every year, approximately 2 million persons in India develop tuberculosis (TB), accounting for one fourth of the world's new TB cases. Organized TB control activities have existed in India for 40 years; however, the quality of diagnosis and treatment of TB in the public and private sectors has been variable, and TB incidence and prevalence trends have not changed substantially over this time. In 1992, the Indian government established a Revised National Tuberculosis Control Programme (RNTCP) using the directly observed treatment, short-course (DOTS) strategy recommended by the World Health Organization (WHO) (3). The DOTS strategy consists of sustained government commitment, effective laboratory-based diagnosis, standard treatment given under direct observation, secure drug supply, and systematic monitoring and evaluation. RNTCP was implemented in pilot areas beginning in 1993; large-scale implementation of the program began in late 1998. This report summarizes the process, outcomes, and challenges of RNTCP in India. RNTCP has implemented DOTS rapidly and has yielded positive results in TB control; however, continued commitment from Indian government authorities and the international community is needed to sustain and expand this ongoing program. PMID- 11925020 TI - Tuberculosis outbreak on an American Indian reservation--Montana, 2000-2001. AB - During May 2000-January 2001, five tuberculosis (TB) cases, linked by contact and DNA fingerprinting, were reported from the Fort Belknap Indian Reservation in rural Montana. Before this, only one case of TB had been reported from the reservation since 1992. To determine the cause of the outbreak, the Fort Belknap Tribal Health Department and the Indian Health Service (IHS) conducted an investigation and requested assistance from the Montana State Department of Public Health and Human Services (DPHHS) and CDC to improve case finding and medical management of persons with TB. This report summarizes the results of the investigation and demonstrates how, in low incidence areas, rapid expansion of local capacity for TB control is critical to eliminate TB in the United States. PMID- 11925021 TI - Progress toward elimination of Haemophilus influenzae type b invasive disease among infants and children--United States, 1998-2000. AB - Haemophilus influenzae type b (Hib) was the leading cause of bacterial meningitis and a major cause of other serious invasive diseases among children aged <5 years in the United States before Hib conjugate vaccines became available in 1988. In 1991, all infants starting at age 2 months were recommended to receive Hib conjugate vaccines; by 1996, incidence of Hib invasive disease (i.e., illness clinically compatible with invasive disease, such as meningitis or sepsis, with isolation of the bacterium from a normally sterile site) among children aged <5 years had declined by >99%. This report presents 1998-2000 Haemophilus influenzae (Hi) surveillance data, which indicate that the incidence of reported Hib invasive disease remains low. Achieving the national health objective for 2010 of reducing to zero indigenous Hib invasive disease cases in children aged <5 years will require improved age-appropriate vaccination of children, complete reporting of vaccination and relevant medical histories, standardization of the serotyping procedure, and complete ascertainment and reporting of serotype for all Hi invasive disease cases. PMID- 11925022 TI - Cardiac xenotransplantation: ethics and potential application. PMID- 11925023 TI - Attempts to prepare suitable complement regulatory molecules for clinical xenotransplantation. PMID- 11925024 TI - The immunological hurdles to cardiac xenotransplantation. AB - The main hurdle to clinical application of cardiac xenotransplantation is the immune response of the recipient against the graft. Although all xenografts arouse an intense immune response, the effect of that response depends very much on whether the graft consists of isolated cells or an intact organ, such as the heart. Intact organs, which are transplanted by primary vascular anastomosis, are subject to severe vascular injury owing to the reaction of immune elements with the endothelial lining of donor blood vessels. Vascular injury leads to hyperacute rejection, acute vascular rejection, and chronic rejection. The immunological basis for these types of rejection and potential therapies, which might be used to avert them, are discussed. PMID- 11925025 TI - Genetic engineering for xenotransplantation. AB - Xenotransplantation is being pursued vigorously to solve the shortage of allogeneic donor organs. Experimental studies of the major xenoantigen (Gal) and of complement regulation enable model xenografts to survive hyperacute rejection. When the Gal antigen is removed or reduced and complement activation is controlled, the major barriers to xenograft survival include unregulated coagulation within the graft and cellular reactions involving macrophages, neutrophils, natural killer (NK) cells, and T lymphocytes. Unlike allografts, where specific immune responses are the sole barrier to graft survival, molecular differences between xenograft and recipient that affect normal receptor-ligand interactions (largely active at the cell surface and which may not be immunogenic), are also involved in xenograft failure. Transgenic strategies provide the best options to control antigen expression, complement activation, and coagulation. Although the Gal antigen can be eliminated by gene knockout in mice, that outcome has only become a possibility in pigs due to the recent cloning of pigs after nuclear transfer. Instead, the use of transgenic glycosyl transferase enzymes and glycosidases, which generate alternative terminal carbohydrates on glycolipids and glycoproteins, has reduced antigen in experimental models. As a result, novel strategies are being tested to seek the most effective solution. Transgenic pigs expressing human complement-regulating proteins (DAF/CD55, MCP/CD46, or CD59) have revealed that disordered regulation of the coagulation system requires attention. There will undoubtedly be other molecular incompatibilities that need addressing. Xenotransplantation, however, offers hope as a therapeutic solution and provides much information about homeostatic mechanisms. PMID- 11925026 TI - On-pump and off-pump coronary artery bypass grafting in the elderly: predictors of adverse outcome. AB - OBJECTIVE: To establish the role that coronary artery bypass grafting (CABG) without cardiopulmonary bypass (CPB) may have in improving perioperative outcomes of patients 70 years of age and older. BACKGROUND: Coronary revascularization in elderly patients is associated with morbidity and mortality rates higher than those observed in younger patients. The impact of CABG without CPB on perioperative outcomes has not been clearly established. METHODS: This retrospective, nonrandomized study consisted of 1,872 CABG patients. Of these, 1389 underwent CABG with CPB (CPB group) and 483 patients underwent CABG without CPB (off-pump group). Preoperative variables and outcomes were compared between the two groups. Multivariate logistic regression analysis was used to identify independent predictors of mortality, stroke, and adverse outcome. RESULTS: Demographics, Canadian Cardiovascular Society staging, operative priority, and other preoperative variables were comparable between the two groups. The prevalence of previous myocardial infarction was higher in the CPB group (62.6% vs 56.7%; p < 0.005), whereas the prevalence of calcified aorta and preoperative renal failure were higher in the off-pump group (5.4% vs 9.5%; p = 0.04 and 1.7% vs 3.3%; p = 0.04, respectively). Although the graft/patient ratio was higher in the CPB group (3.4 vs 1.9), these patients displayed more extensive coronary artery involvement. At univariate analysis, patients in the off-pump group had a higher rate of freedom from complications (88.2% vs 81.3%; p < 0.005) and a lower incidence of stroke (2.1% vs 4.2%; p = 0.034) than patients in the CPB group. Although there was a trend for a higher actual mortality in the off-pump group (4.8% vs 3.7%; p = ns), the risk adjusted mortality in this group was lower (1.9% vs 2.1%). Multivariate analysis showed that while the use of CPB correlated independently with an increased risk of overall complications, it was not associated with a higher probability of death or stroke. CONCLUSIONS: This investigation suggests that elderly patients undergoing CABG may benefit from off pump revascularization, as the use of CPB correlated independently with an increased risk of overall complications. However, CPB did not emerge as an independent predictor of death or stroke at multivariate analysis. PMID- 11925027 TI - The anticalcific effect of glutaraldehyde detoxification on bioprosthetic aortic wall tissue in the sheep model. AB - BACKGROUND: Increasing concentrations of glutaraldehyde (GA) lead to a decreased rather than increased calcification of bioprosthetic aortic wall tissue. This study determined to what extent the benefit of better cross-linking is masked by the intrinsic propensity of GA towards calcification. MATERIALS AND METHODS: Porcine aortic roots were immediately fixed at the abattoir at three different concentrations of GA (0.2%, 1.0%, and 3.0% for 1 week at 4 degrees C). Subsequently, roots underwent a GA extraction process using high volumes of Urazole solution (acetic acid buffer, pH 4.5, 37 degrees C, 1 week) followed by NaBH4 reduction (2 days, 37 degrees C). Roots were implanted in the distal aortic arch of young sheep for 6 weeks and 6 months. Calcium analysis was quantitatively done by atomic absorption spectrophotometry and qualitatively assessed by light microscopy on Von Kossa stains. RESULTS: There was a distinct anticalcification effect of GA detoxification after 6 weeks (56.8% to 97.9%; 95% confidence interval [CI]), which stabilized on a more moderate level after 6 months of implantation (19.1% to 31.6%; 95% CI). The most pronounced effect of GA extraction was seen in 0.2% fixed tissue, where aortic wall calcification was mitigated by 97% and 32% after 6 weeks and 6 months, respectively. Mitigation of aortic wall calcification was 71% (6 weeks) and 21% (6 months) in the 3.0% GA group. The combined effect of higher cross-link density and detoxification achieved an 82% (6 weeks) and 48% (6 months) reduction of calcium levels in the 3.0% GA group. In long-term implants (6 months), detoxification alone on top of standard 0.2% GA fixation was as effective (from 174.1 +/- 11.9 microg/mg without detoxification to 119.3 +/- 19.3 microg/mg with detoxification) as 3.0% fixation (114.8 +/- 10.0 microg/mg without detoxification to 91.3 +/- 11.5 microg/mg with detoxification). CONCLUSION: We were able to determine in the circulatory sheep model to what degree the intrinsic procalcific effect of GA counteracts the protective effect of higher cross-link density. Our study also established that the effect of detoxification is particularly pronounced in commercial low-grade fixation. PMID- 11925029 TI - Minimally invasive, nonendoscopic saphenectomy for coronary bypass surgery. AB - In 200 consecutive patients who underwent elective and emergency coronary bypass surgery, a minimally invasive, nonendoscopic technique for greater saphenous vein harvesting was used and the clinical results evaluated. The technique is easy, rapid, and routinely applicable in elective and emergency coronary bypass procedures. The cosmetic effect is excellent and the saphenectomy-related morbidity low. PMID- 11925028 TI - Should mild-to-moderate and moderate ischemic mitral regurgitation be corrected in patients with impaired left ventricular function undergoing simultaneous coronary revascularization? AB - INTRODUCTION: Mitral valve regurgitation (MR) occurring as a result of myocardial ischemia and global left ventricular (LV) dysfunction predicts poor outcome. This study assessed the feasibility of mitral valve (MV) surgery concomitant with coronary artery bypass grafting (CABG) in patients with mild-to-moderate and moderate ischemic MR and impaired LV function. MATERIALS AND METHOD: From January 1996 to July 2000, 49 patients (group 1) and 50 patients (group 2) with grade II and grade III ischemic MR and LV ejection fraction (EF) between 17% and 30% underwent combined MV surgery and CABG (group 1) or isolated CABG (group 2). LVEF (%), LV end-diastolic diameter (EDD) (mm), LV end-diastolic pressure (EDP) (mmHg), and LV end-systolic diameter (ESD) (mm) were 27.5 +/- 5, 67.7 +/- 7,27.7 +/- 4, and 51.4 +/- 7, respectively in group 1 versus 27.8 +/- 4, 67.5 +/- 6, 27.5 +/- 5, and 51.2 +/- 6, respectively in group 2. Groups 1 and 2 were divided into Groups 1A and 2A with mild-to-moderate MR (22 [45%] and 28 [56%] patients, respectively) and groups 1B and 2B with moderate MR (27 [55%] and 22 [46%], respectively). In group 1, MV repair was performed in 43 (88%) patients and MV replacement in 6 (12%) patients. RESULTS: Preoperative data analysis did not reveal any difference between groups. Five (10%) patients in group 1 died versus 6 (12%) in group 2 (p = ns). Within 6 months after surgery, LV function and its geometry improved significantly in group 1 versus group 2 (LVEF, p < 0.001; LVEDD, p = 0.002; LVESD, p = 0.003; and LVEDP (p < 0.001) improved significantly in group 1 instead of a mild improvement in Group 2). The regurgitation fraction decreased significantly in group 1 patients after surgery (p < 0.001). There was an inverse strong correlation between postoperative forward cardiac output and regurgitation fraction (p < 0.001). LVEF and LVESD improved significantly in group 1 versus group 2 patients (p = 0.04 and p = 0.02, respectively). The cardiac index increased significantly in group 1 and 2 (p < 0.001 and p = 0.03, respectively). LV function and geometry improved significantly postoperatively in group 1B versus group 2B (LVEDD, p = 0.027; LVESD, p = 0.014; LVEDP, p = 0.034; and LVEF, p = 0.02), instead of a mild improvement in group 1A versus group 2A (LVESD, p = 0.015; LVEF, p = 0.046; and LVEDD and LVEDP, p = 0.05). At follow-up, 4 (67%) of 6 patients undergoing MV replacement died versus 5 (11.5%) of 43 patients undergoing MV repair in group 1 (p = 0.007). The overall survival at 3 years in Group 2 was significantly lower than group 1 (p < 0.009). CONCLUSION: MV repair and replacement-preserving subvalvular apparatus in patients with impaired LV function offered acceptable outcomes in terms of morbidity and survival. Surgical correction of mild-to-moderate and moderate MR in patients with impaired LV function should be taken into consideration since it yields better survival and improved LV function. PMID- 11925030 TI - Retropericardial hematoma: pitfall in beating heart coronary surgery. AB - Beating heart coronary surgery, now an accepted technique of myocardial revascularization, is perceived to have fewer complications than the conventional on-pump approach. However, little attention has been paid to complications that may be specific to the off-pump method. We describe the unusual development of a large retropericardial hematoma following otherwise uneventful beating heart coronary surgery that resulted in global cardiac displacement. This was recognized and investigated promptly, and led ultimately to complete resolution without morbidity with expectant treatment alone. The implication of this observation is discussed along with other reports of perioperative complications following beating heart coronary surgery. PMID- 11925031 TI - Clinical applications of cardiovascular angiogenesis. AB - Angiogenesis is fundamental to both normal physiologic (wound healing) and pathologic (cancer) processes. Manipulation of divergent angiogenic signals promises effective therapy of atherosclerotic cardiovascular disease. Positive proangiogenic strategies promise collateral circulation to ischemic territories, while negative antiangiogenic strategies starve the fibromuscular proliferation within the atherosclerotic lesion. Indeed, recent phase 1 trials suggest that delivering DNA or recombinant protein to the site of vascular occlusion may stimulate physiologically significant collateral circulation in chronically ischemic myocardium. While symptomatic and functional improvement has been documented, toxicity profiles and effects on long-term patient survival are still unclear. The purposes of this article are as follows: (1) to review the pathophysiologic basis for pro- and antiangiogenic strategies in the treatment of cardiovascular disease, (2) to examine the clinical trials of proangiogenic gene or recombinant protein delivery into ischemic beds, and conversely, (3) to explore antiangiogenic strategies in the prevention and treatment of intimal neovascularization and smooth muscle proliferation within the vessel wall. PMID- 11925032 TI - Jasmonate-related mutants of Arabidopsis as tools for studying stress signaling. AB - Jasmonates are naturally occurring signal compounds that regulate plant growth and development, and are involved in plant responses to several environmental stress factors. The mode of action of jasmonates has been investigated traditionally by analysis of the effects of exogenous application of these compounds, including identification of jasmonate-responsive genes and determination of their expression and responsive promoter elements. In addition, jasmonate biosynthesis has been studied by identification of biosynthetic enzymes, use of inhibitors and determination of endogenous jasmonate levels. Recently, several mutants defective in jasmonate biosynthesis and signaling have been isolated and their phenotypes shed new light on the role of jasmonates and jasmonate signaling in plant responses to pathogens, insects and ozone. PMID- 11925033 TI - An auxin surge following fertilization in carrots: a mechanism for regulating plant totipotency. AB - All plants exhibit the property of cellular totipotency, whereby individual cells can regenerate into an entire organism. However little is known about the underlying mechanisms regulating totipotency. Using a preparative microtechnique, we report an 80-fold surge in the concentration of free auxin that is correlated with the initial stages of zygotic embryogenesis in carrots. The concentration of free IAA increases from a basal level of ca. 25 ng/g FW in unfertilized ovules to ca. 2.000 ng/g FW in the late globular and early heart stages, then back to the basal level in the torpedo stage. This initial increase in IAA levels is diagnostic of the activity of the tryptophan-mediated pathway for IAA biosynthesis, while the maintenance of the basal levels is attributed to the tryptophan-independent pathway for IAA biosynthesis. Our observations are consistent with the hypothesis that the sequential activation of alternative IAA biosynthetic pathways is a critical mechanism for regulating carrot (Daucus carota L. cv. Danvers 126) embryogenesis and other instances of plant totipotency. PMID- 11925034 TI - Antisense repression of cytosolic phosphoglucomutase in potato (Solanum tuberosum) results in severe growth retardation, reduction in tuber number and altered carbon metabolism. AB - The aim of this work was to investigate the role of cytosolic phosphoglucomutase (PGM; EC 5.4.2.2) in the regulation of carbohydrate metabolism. Many in vitro studies have indicated that PGM plays a central role in carbohydrate metabolism; however, until now the importance of this enzyme in plants has not been subject to reverse-genetics investigations. With this intention we cloned the cytosolic isoform of potato PGM (StcPGM) and expressed this in the antisense orientation under the control of the CaMV 35 S promoter in potato plants. We confirmed that these plants contained reduced total PGM activity and that loss in activity was due specifically to a reduction in cytosolic PGM activity. These plants were characterised by a severe phenotype: stunted aerial growth combined with limited root growth and a reduced tuber yield. Analysis of the metabolism of these lines revealed that leaves of these plants were inhibited in sucrose synthesis whereas the tubers exhibited decreased levels of sucrose and starch as well as decreased levels of glycolytic intermediates but possessed unaltered levels of adenylates. Furthermore, a broader metabolite screen utilising GC-MS profiling revealed that these lines contained altered levels of several intermediates of the TCA cycle and of amino acids. In summary, we conclude that cytosolic PGM plays a crucial role in the sucrose synthetic pathway within the leaf and in starch accumulation within the tuber, and as such is important in the maintenance of sink-source relationships. PMID- 11925035 TI - Structure and development of epiphylly in knox-transgenic tobacco. AB - Anatomical modifications and developmental patterns of tobacco (Nicotiana tabacum L.) plants transgenic for the barley (Hordeum vulgare L.) homeo box genes bkn-1 or bkn-3 were analysed and the morphogenetic processes interpreted. No appreciable difference between bkn-1 and bkn-3 transgenic tobacco was observed in the examined transgenic plants. They were dwarfed and developed epiphyllous appendages on all leaves except the cotyledons. Two major categories of epiphyllous structure were recognised: (i) vegetative shoots on juvenile heart shaped leaves and (ii) inflorescences or flowers on adult lanceolate leaves. Both types of epiphyllous structure originated in the subepidermal layers, either directly from individual meristems or from an expanded meristematic "cushion". Structural analysis confirmed that the occurrence of epiphylls in the tobacco transformants represented a switch from determinate to indeterminate leaf growth and reproduced the phenotype of the barley Hooded mutant, which had earlier been shown to be associated with the bkn-3 gene [K.J. Muller et al. (1995) Nature 374:727-730]. PMID- 11925036 TI - Myosin, microtubules, and microfilaments: co-operation between cytoskeletal components during cambial cell division and secondary vascular differentiation in trees. AB - The immunolocalisation of unconventional myosin VIII ('myosin') in the cells of the secondary vascular tissues of angiosperm (Populus tremula L. x P. tremuloides Michx. and Aesculus hippocastanum L.) and gymnosperm (Pinus pinea L.) trees is described for the first time and related to other cytoskeletal elements, as well as to callose. Both myosin and callose are located at the cell plate in dividing cambial cells, whereas actin microfilaments are found alongside the cell plate; actin and tubulin are both associated with the phragmoplast. Myosin and callose also localise to the plasmodesmata-rich pit fields in the walls of living cells, which are particularly abundant within the common walls between ray cells and between ray cells and axial parenchyma cells in the phloem and xylem. In those xylem ray cells that contact developing vessel elements and tracheids, myosin, tubulin, actin and callose are localised at the periphery of developing contact and cross-field pits; the respective antibodies also highlight the bordered pits between vessels and between tracheids. The aperture of the bordered pits, whose diameter diminishes as the over-arching border of these pits develops, also houses myosin, actin and tubulin. Myosin, actin and callose are also found together around the sieve pores of sieve elements and sieve cells. We suggest that an acto-myosin contractile system (a 'plant muscle') is present at the cell plate, the sieve pores, the plasmodesmata within the walls of long-lived parenchyma cells, and at the apertures of bordered pits during their development. PMID- 11925038 TI - Characterization and modelling of the hydrophobic domain of a sunflower oleosin. AB - The oleosins are a group of hydrophobic proteins present on the surface of oil bodies in seeds, where they are thought to prevent coalescence. They contain a central hydrophobic domain of 68-74 residues that is thought to form a loop into the triacylglycerol matrix of the oil body, but the conformation adopted by this sequence is uncertain. We have therefore expressed an oleosin cDNA from sunflower (Helianthus annuus L.) in Escherichia coli as a fusion with maltose-binding protein (MBP) and isolated a peptide corresponding to the hydrophobic domain by sequential digestion with factor Xa (to remove the MBP) followed by trypsin and Staphylococcus V8 protease to remove the N- and C-terminal domains of the oleosin. Circular dichroism spectroscopy of the peptide in two solvent systems chosen to mimic the environment within the oil body (trifluoroethanol and SDS) demonstrated high proportions of alpha-helical structure, with no beta-sheet. A model was therefore developed in which the domain forms an alpha-helical hairpin structure, the two helices being separated by a turn region. We consider that this model is consistent with our current knowledge of oleosin structure and properties. PMID- 11925037 TI - A critical role for ethylene in hydrogen peroxide release during programmed cell death in tomato suspension cells. AB - Camptothecin, a topo isomerase-I inhibitor used in cancer therapy, induces apoptosis in animal cells. In tomato (Lycopersicon esculentum Mill.) suspension cells, camptothecin induces cell death that is accompanied by the characteristic nuclear morphological changes such as chromatin condensation and nuclear and DNA fragmentation that are commonly associated with apoptosis in animal systems. These effects of camptothecin can effectively be blocked by inhibitors of animal caspases, indicating that, in tomato suspension cells, camptothecin induces a form of programmed cell death (PCD) with similarities to animal apoptosis (A.J. De Jong et al. (2000) Planta 211:656-662). Camptothecin induced cell death was employed to study processes involved in plant PCD. Camptothecin induced a transient increase in H2O2 production starting within 2 h of application. Both camptothecin-induced cell death and the release of H2O2 were effectively blocked by application of the calcium-channel blocker lanthanum chloride, the caspase specific inhibitor Z-Asp-CH2-DCB, or the NADPH oxidase inhibitor diphenyl iodonium, indicating that camptothecin exerts its effect on cell death through a calcium- and caspase-dependent stimulation of NADPH oxidase activity. In addition, we show that ethylene is an essential factor in camptothecin-induced PCD. Inhibition of either ethylene synthesis or ethylene perception by L-alpha-(2 aminoethoxyvinyl)glycine or silver thiosulphate, respectively, blocked camptothecin-induced H2O2 production and PCD. Although, in itself, insufficient to trigger H2O2 production and cell death, exogenous ethylene greatly stimulated camptothecin-induced H2O2 production and cell death. These results show that ethylene is a potentiator of the camptothecin-induced oxidative burst and subsequent PCD in tomato cells. The possible mechanisms by which ethylene stimulates cell death are discussed. PMID- 11925039 TI - Biochemical and morphological characterization of sulfur-deprived and H2 producing Chlamydomonas reinhardtii (green alga). AB - Sulfur deprivation in green algae causes reversible inhibition of photosynthetic activity. In the absence of S, rates of photosynthetic O2 evolution drop below those of O2 consumption by respiration. As a consequence, sealed cultures of the green alga Chlamydomonas reinhardtii become anaerobic in the light, induce the "Fe-hydrogenase" pathway of electron transport and photosynthetically produce H2 gas. In the course of such H2-gas production cells consume substantial amounts of internal starch and protein. Such catabolic reactions may sustain, directly or in directly, the H2-production process. Profile analysis of selected photosynthetic proteins showed a precipitous decline in the amount of ribulose-1,5-bisphosphate carboxylase-oxygenase (Rubisco) as a function of time in S deprivation, a more gradual decline in the level of photosystem (PS) II and PSI proteins, and a change in the composition of the PSII light-harvesting complex (LHC-II). An increase in the level of the enzyme Fe-hydrogenase was noted during the initial stages of S deprivation (0-72 h) followed by a decline in the level of this enzyme during longer (t >72 h) S-deprivation times. Microscopic observations showed distinct morphological changes in C. reinhardtii during S deprivation and H2 production. Ellipsoid-shaped cells (normal photosynthesis) gave way to larger and spherical cell shapes in the initial stages of S deprivation and H2 production, followed by cell mass reductions after longer S-deprivation and H2 production times. It is suggested that, under S-deprivation conditions, electrons derived from a residual PSII H2O-oxidation activity feed into the hydrogenase pathway, thereby contributing to the H2-production process in Chlamydomonas reinhardtii. Interplay between oxygenic photosynthesis, mitochondrial respiration, catabolism of endogenous substrate, and electron transport via the hydrogenase pathway is essential for this light-mediated H2-production process. PMID- 11925040 TI - Composition and content of glucosinolates in developing Arabidopsis thaliana. AB - The glucosinolate composition and content in various tissues of Arabidopsis thaliana (L.) Heynh. ecotype Columbia during development from seeds to bolting plants were determined in detail by high-performance liquid chromatography. Comparison of the glucosinolate profiles of leaves, roots and stems from mature plants with those of green siliques and mature seeds indicated that a majority of the seed glucosinolates were synthesized de novo in the silique. A comparison of the glucosinolate profile of mature seeds with that of cotyledons indicated that a major part of seed glucosinolates was retained in the cotyledons. Turnover of glucosinolates was studied by germination of seeds containing radiolabelled p hydroxybenzylglucosinolate (p-OHBG). Approximately 70% of the content of [14C]p OHBG in the seeds was detected in seedlings at the cotyledon stage and [14C]p OHBG was barely detectable in young plants with rosettes of six to eight leaves. The turn-over of p-OHBG was found to coincide with the expression of the glucosinolate-degrading enzyme myrosinase, which was detectable at very low levels in seedlings at the cotyledon stage, but which dramatically increased in leaves from plants at later developmental stages. This indicates that there is a continuous turnover of glucosinolates during development and not only upon tissue disruption. PMID- 11925041 TI - The growth defect of lrt1, a maize mutant lacking lateral roots, can be complemented by symbiotic fungi or high phosphate nutrition. AB - The growth of three maize (Zea mays L.) mutants, each impaired in the formation of one individual element of its root system, was compared under "natural" limiting phosphate conditions (0.1 mM). Mutant plants exhibiting a reduction in root hairs (rth3-1) or a depletion of crown and brace roots (rtcs) grew as well as the corresponding wild-type plants. However, mutant plants lacking lateral roots (lrt1) showed a strong reduction in plant growth. The growth defect of lrt1 was overcome when it was grown in association with an arbuscular mycorrhizal fungus, Glomus mosseae. Establishment of symbiosis was associated with the occurrence of a new type of lateral root. These new lateral roots were stunted and highly branched, giving rise to a bush-like structure. Supply of high phosphate (1 microM) ameliorated the growth of lrt1 plants too, but less efficiently than the symbiosis did. Hence, arbuscular mycorrhizal fungi as well as phosphate functionally complemented the lrt1 mutation. PMID- 11925042 TI - Molecular and physiological characterization of arabidopsis GAI alleles obtained in targeted Ds-tagging experiments. AB - Bioactive gibberellin (GA) is an essential regulator of vascular plant development. The GAI gene of Arabidopsis thaliana (L.) Heynh. encodes a product (GAI) that is involved in GA signalling. The dominant mutant gai allele encodes an altered product (gai) that confers reduced GA responses, dwarfism, and elevated endogenous GA levels. Recessive, presumed loss-of-function alleles of GAI confer normal height and resistance to the GA biosynthesis inhibitor paclobutrazol. One explanation for these observations is that GAI is a growth repressor whose activity is opposed by GA, whilst gai retains a constitutive repressor activity that is less affected by GA. Previously, we described gai-t6, a mutant allele which contains an insertion of a maize Ds transposable element into gai. Here we describe the molecular and physiological characterization of two further alleles (gai-t5, gai-t7) identified during the Ds mutagenesis experiment. These alleles confer paclobutrazol resistance and normal endogenous GA levels. Thus the phenotype conferred by gai-t5, gai-t6 and gai-t7 is not due to elevated GA levels, but is due to loss of gai, a constitutively active plant growth repressor. PMID- 11925043 TI - Glyoxysomal acetoacetyl-CoA thiolase and 3-oxoacyl-CoA thiolase from sunflower cotyledons. AB - Following chromatography on hydroxyapatite, the elution profile of the thiolase activity of the glyoxysomal fraction from sunflower (Helianthus annuus L.) cotyledons exhibited two peaks when the enzyme activity was assayed with acetoacetyl-CoA as substrate. Only one of these two activity peaks was detectable when a long-chain thiolase substrate was used in the activity assay. The proteins (thiolase I and thiolase II) underlying the two activity peaks detected with acetoacetyl-CoA were of glyoxysomal origin. They were purified using glyoxysomal matrices as starting material, and biochemically characterized. Thiolase I is an acetoacetyl-CoA thiolase (EC 2.3.1.9) exhibiting activity only towards acetoacetyl-CoA (Km = 11 microM). Its contribution to the total glyoxysomal thiolytic activity towards acetoacetyl-CoA amounted to about 15%. Thiolase II is a 3-oxoacyl-CoA thiolase (EC 2.3.1.16). The activity of the enzyme towards 3 oxoacyl-CoAs increased with increasing chain length of the substrate. Thiolase II exhibited a Km value of 27 microM with acetoacetyl-CoA as substrate. and Km values between 3 and 7 microM with substrates having a carbon chain length from 6 to 16 carbon atoms. The thiolase activity of the glyoxysomes towards acetoacetyl CoA and 3-oxopalmitoyl-CoA exceeded the glyoxysomal butyryl-CoA and palmitoyl-CoA beta-oxidation rates, respectively, by about 10-fold at all substrate concentrations employed (1-15 microM). PMID- 11925044 TI - Plant aging increases oxidative stress in chloroplasts. AB - Aging has received considerable attention in biomedicine, but little is known about the regulatory mechanisms responsible for the aging not associated with senescence in plants. This study provides new insights into the relationship between oxidative stress and plant aging, and points out chloroplasts as one of the target organelles of age-associated oxidative stress in plants. We simultaneously analyzed lipid oxidation, photosynthesis, chlorophyll content, de epoxidation state of the xanthophyll cycle, and levels of chloroplastic antioxidant defenses such as beta-carotene and alpha-tocopherol in leaves of the same age in 1-, 3- and 7-year-old Cistus clusii Dunal plants growing under Mediterranean field conditions. Enhanced formation of malondialdehyde in leaves (2.7-fold) and chloroplasts (2.8-fold), decreased photosynthetic activity (25%), and lower chlorophyll (ca. 20%) and chloroplastic antioxidant defense levels (ca. 25%-85%) were observed in 7-year-old plants, when compared with 1- and 3-year-old plants. The differences observed, which were associated with plant aging, were only noticeable in mature nonsenescing plants (7-year-old plants). No differences were observed between pre-reproductive (1-year-old plants) and young plants (3 year-old plants). This study shows that from a certain age, oxidative stress increases progressively in chloroplasts as plants age, whereas photosynthesis is reduced. The results indicate that the oxidative stress associated with the aging in plants accumulates progressively in chloroplasts, and that the contribution of oxidative stress to aging increases as plants age. PMID- 11925046 TI - Isolation and characterization of lipid in phloem sap of canola. AB - Phloem isolated from canola (Brassica napus L.) stems was found to contain phospholipid, diacylglycerol, triacylglycerol, steryl and wax esters, and comparatively high concentrations of unesterified fatty acids. Indeed, the composition of phloem lipid was markedly different from that of microsomal membranes and cytosol isolated from both leaves and stems. Specifically, phloem lipid consisted predominantly of unesterified fatty acids and was enriched in medium-chain fatty acids, in particular, lauric, myristic and pentadecanoic acids. This unique composition also distinguished phloem lipid from that of well characterized cytosolic lipid particles such as oil bodies found in plant cells. Moreover, levels of medium-chain fatty acids in the phloem increased when canola plants were stressed by exposure to sublethal doses of ultraviolet irradiation. Phloem levels of lauric acid, for example, increased by 11-fold upon treatment with sublethal ultraviolet irradiation. Spherical lipid particles were discernible in isolated phloem sap by electron microscopy, suggesting that the lipid in phloem is in the form of lipid particles. The presence of lipid in phloem may be reflective of long-distance lipid transport in plants, primarily in the form of free fatty acids. PMID- 11925045 TI - Starch biosynthesis from triose-phosphate in transgenic potato tubers expressing plastidic fructose-1,6-bisphosphatase. AB - A full length cDNA clone encoding plastidic fructose-1,6-bisphosphatase (cp FBPase), together with a transit peptide, was isolated from a potato (Solanum tuberosum L.) leaf cDNA library. Potato plants were transformed with the isolated cp-FBPase sequence behind a patatin class I promoter to ensure tuber-specific expression of the enzyme. Plant lines were selected which expressed up to 250 mU (g FW)-1 in the developing tubers, which is 10- to 20-fold the activity found in wild-type tubers. Intact amyloplasts were isolated from in vitro-grown minitubers developed in darkness. Comparison with marker enzymes showed that cp-FBPase activity in transgenic tubers, as well as the low FBPase activity in the wild type tubers, was localised inside the amyloplasts. The intact amyloplasts isolated from both wild-type and transgenic tubers synthesised starch from [U 14C] glucose-6-phosphate. Conversely, only the transgenic tubers expressing cp FBPase showed appreciable synthesis of starch from [U-14C] dihydroxyacetone phosphate, and this synthesis rate was correlated to the activity of cp-FBPase. Thus, the expression of cp-FBPase in tubers allows for a new route of starch biosynthesis from triose-phosphates imported from the cytosol. The transgenic tubers did not differ from wild-type tubers with respect to starch content, or the levels of neutral sugars and phosphorylated hexoses. PMID- 11925047 TI - Phytochelatin synthesis is not responsible for Cd tolerance in the Zn/Cd hyperaccumulator Thlaspi caerulescens (J. & C. Presl). AB - Thlaspi caerulescens (J. & C. Presl, "Prayon") is a heavy-metal hyperaccumulator that accumulates Zn and Cd to high concentrations (40,000 and 4,000 mg kg DW-1 respectively) without phytotoxicity. The mechanism of Cd tolerance has not been characterized but reportedly involves vacuolar sequestration. The role of phytochelatins (PCs) in metal tolerance in T. caerulescens and the related non accumulator T. arvense was examined. Although PCs were produced by both species in response to Cd, these peptides do not appear to be involved in metal tolerance in the hyperaccumulator. Leaf and root PC levels for both species showed a similar positive correlation with tissue Cd, but total PC levels in the hyperaccumulator were generally lower, despite correspondingly higher metal concentrations. The lack of a role for PCs in the hyperaccumulator's response to metal stress suggests that other mechanisms are responsible Cd tolerance. The lower level of leaf PCs in T. caerulescens also implies that Cd in the shoot is sequestered in a compartment or form that does not elicit a PC response. PMID- 11925048 TI - Expression of the pea S-adenosylmethionine decarboxylase gene is involved in developmental and environmental responses. AB - A cDNA, able to complement the S-adenosyl-L-methionine decarboxylase (SAMdC; EC 4.1.1.50)-defective yeast strain Y342, has been isolated from pea (Pisum sativum L.). Expression of the SAMdC gene was characterised during pea development. Northern analysis showed a differential expression of the pea SAMdC gene in vegetative and reproductive tissues. The highest SAMdC mRNA levels were found in undifferentiated callus and tissues with high rates of cell division, and at the onset of fruit development. SAMdC expression was also induced in senescing ovaries, probably in relation to an accumulation of spermine during ovary senescence. Finally, the levels of SAMdC transcripts in leaves and shoots were differentially affected by ozone treatment. PMID- 11925049 TI - Evidence for regulation of resistance in Arabidopsis to Egyptian cotton worm by salicylic and jasmonic acid signaling pathways. AB - Signaling cross-talk between wound- and pathogen-response pathways influences resistance of plants to insects and disease. To elucidate potential interactions between salicylic acid (SA) and jasmonic acid (JA) defense pathways, we exploited the availability of characterized mutants of Arabidopsis thaliana (L.) Heynh. and monitored resistance to Egyptian cotton worm (Spodoptera littoralis Boisd.; Lepidoptera: Noctuidae). This generalist herbivore is sensitive to induced plant defense pathways and is thus a useful model for a mechanistic analysis of insect resistance. As expected, treatment of wild-type Arabidopsis with JA enhanced resistance to Egyptian cotton worm. Conversely, the coil mutant, with a deficiency in the JA response pathway, was more susceptible to Egyptian cotton worm than wild-type Arabidopsis. By contrast, the nprl mutant, with defects in systemic disease resistance, exhibited enhanced resistance to Egyptian cotton worm. Pretreatment with SA significantly reduced this enhanced resistance of nprl plants but had no influence on the resistance of wild-type plants. However, exogenous SA reduced the amount of JA that Egyptian cotton worm induced in both npr1 mutant and wild-type plants. Thus, this generalist herbivore engages two different induced defense pathways that interact to mediate resistance in Arabidopsis. PMID- 11925050 TI - Mycorrhizal colonization of transgenic aspen in a field trial. AB - Mycorrhizal colonization of genetically modified hybrid aspen (Populus tremula x P. tremuloides Michx.) was investigated over 15 months in a field experiment. The aspen carried the rolC gene from Agrobacterium rhizogenes under control of either the constitutive cauliflower mosaic virus 35S promoter or the light-inducible rbcS promoter. Arbuscular mycorrhizas (AMs) were rare in all root samples, while fully developed ectomycorrhizas (EMs) were found in all samples. No significant differences in the degree of mycorrhizal colonization between aspen lines were seen with either AMs or EMs. The EM community on the release area was dominated by four fungal species that formed more than 90% of all mycorrhizas, while eleven EM types were found occasionally. Mycorrhizal diversity did not differ between transgenic and non-transgenic trees. The structure of mycorrhizal communities was similar for most aspen lines. The sole significant difference was found in the abundance and development of one of the four common EM morphotypes, which was rare and poorly developed on roots from the transgenic aspen line Esch5:35S-rolC #5 compared with non-transgenic controls. This effect is clone specific as the formation of this EM type was not affected by the transgene expression in the other transgenic line, Esch5:35S-rolC-#1. This is the first demonstration of a clonal effect influencing the ability of a transgenic plant to form a mycorrhizal symbiosis with a potential fungal partner. PMID- 11925052 TI - Effects of self-administered alcohol or sucrose preloads on subsequent consumption in the rat. AB - OBJECTIVE: The initial drink of alcohol is often conceptualized as "priming" the individual for the following drinking bout. For the alcoholic, this priming effect has been considered a key for the loss of control that then occurs. Although there have been a few animal studies examining the effects of an investigator-administered ethanol preload on subsequent ethanol self administration, the effects of a small self-administered oral preload on subsequent consumption have not been examined. METHOD: Adult, male rats, initiated to self-administer ethanol using the sucrose-substitution procedure, were given brief access periods to drink ethanol or water, 5 minutes prior to a second opportunity to press a lever for an additional 20-minute access to a 10% ethanol solution. A second group of rats were trained to press a lever to gain access to a 3% sucrose solution, and the effects of sucrose or water preloads were examined and compared with results in the ethanol group. RESULTS: In the ethanol group, both the ethanol and water preload intakes increased as preload access time increased and were not different from each other. However, ethanol preloads at the longer access times (60 seconds and 120 seconds) decreased subsequent ethanol consumption and at the highest time also affected ethanol seeking behavior. Equal volumes of water intake at these longer access times had no effects on subsequent ethanol consumption. In the sucrose group, sucrose preload intakes increased as access time increased, but water preload intakes did not. Neither sucrose nor water preloads had any effect on subsequent sucrose consumption. CONCLUSIONS: The data failed to find any priming effect of ethanol preloads in terms of increased subsequent ethanol consumption. It appears that a major factor in the regulation of ethanol intake for the rat in this training procedure is the postingestional effects of ethanol, because taste stimuli did not appear to be important. However, it appears that these ethanol postingestive stimuli are not identical to those involved in the regulation of sucrose consumption. PMID- 11925051 TI - Effects of tiagabine and diazepam on operant ethanol self-administration in the rat. AB - OBJECTIVE: Benzodiazepines (BDZ) are widely used in the treatment of anxiety and ethanol withdrawal. It has been suggested that this class of compounds may increase the reinforcing value of ethanol; however, the literature is scarce. Tiagabine has recently been introduced into clinical use as an anti-epileptic drug. It acts through inhibiting gamma-aminobutyric acid (GABA) reuptake, and thus represents a pharmacodynamically novel principle for potentiating GABAergic transmission. The objective of the present study was to examine whether these two manners of modulating GABAergic transmission would affect ethanol self administration in rats. METHOD: Rats were trained on an operant oral ethanol self administration task in a two-lever free-choice paradigm. When trained, subjects were treated with tiagabine (2, 6 and 18 mg/kg, intraperitoneally [i.p.]) or diazepam (0.5, 1.5 and 4.5 mg/kg, i.p.). Postsession blood alcohol concentrations and locomotor activity measures also were obtained. RESULTS: At nonsedating doses, neither tiagabine nor diazepam affected operant ethanol self administration. At the highest doses (18 and 4.5 mg/kg, respectively), both drugs suppressed ethanol self-administration but also induced significant suppression of locomotion, indicative of sedation. CONCLUSIONS: Systemic administration of either the GABA-uptake blocker, tiagabine, or the GABA/BDZ agonist, diazepam, at nonsedating doses does not seem to affect oral ethanol self-administration. PMID- 11925053 TI - Decomposing the relationships between pretreatment social network characteristics and alcohol treatment outcome. AB - OBJECTIVE: In Project MATCH a summary measure of network support for drinking identified matching and prognostic effects. The goals of the present analyses were (1) to determine which of the 11 component indexes are most predictive of treatment outcome regardless of treatment type and (2) to determine which of the indexes are most influential in the already demonstrated network support by treatment interaction effect. METHOD: This is a secondary data analysis of the outpatient arm of Project MATCH (N = 952), focusing primarily on the Important People and Activities instrument administered pretreatment. RESULTS: Patients with larger daily networks and patients with more abstainers/recovering alcoholics in their networks had a better prognosis. Patients with a higher network drinking frequency did better in Twelve Step Facilitation than in Motivational Enhancement Therapy. CONCLUSIONS: Three of the 11 indexes can be used to replicate the prognostic and matching effects found for an overall index of network support. These may be measured by a short form of the Important People and Activities instrument. PMID- 11925054 TI - Psychiatric diagnoses among Mission Indian children with and without a parental history of alcohol dependence. AB - OBJECTIVE: The primary goal was to determine whether Mission Indian children of alcoholics (COAs) have higher rates of psychiatric disorders than Mission Indian non-COAs. A secondary goal was to explore associations of other familial variables with rates of childhood psychiatric disorders. METHOD: Psychiatric diagnoses were obtained from the primary caregiver, using the Semi-Structured Assessment for the Genetics of Alcoholism (SSAGA), for 117 (63 female) Mission Indian children; 81 of the children were interviewed directly. Of the 117 subjects, 92 (79%) were COAs and 25 (21%) were not. RESULTS: There were no differences in rates of psychiatric disorders between COAs and non-COAs. Living in a household without a mother was associated with a higher rate of separation anxiety disorder, and living in a household without a father was associated with a higher rate of both oppositional defiant disorder and separation anxiety disorder. CONCLUSIONS: Parental alcohol dependence does not appear to relate to increased psychopathology among Mission Indian children, but living in a single parent household may be associated with increased rates of some psychiatric disorders. PMID- 11925055 TI - Drinking-driving as a component of problem driving and problem behavior in young adults. AB - OBJECTIVE: This study replicated and extended a conceptual model of drinking driving and its relationship to other problem behavior to determine (1) if the model characterized the problem driving and problem behavior of young adults from the general population and (2) if the model applied to women as well as to men. METHOD: Telephone survey data (measures of drinking-driving, drug-driving, risky driving, problem drinking, drug use and delinquent behavior) collected from young adults (N = 4,230, 53% female) were used in structural equation modeling. Two models were developed--one for problem driving and one for problem behavior--each testing the structural associations among latent variables and testing them as latent indicators of common second-order latent variables. RESULTS: The results supported the hypothesis that drinking-driving (along with problem drinking, drug use and delinquent behavior) is an indicator of problem behavior in the general population sample. Also, drinking-driving, drug-driving and risky driving were demonstrated to be indicators of a common construct: problem driving. The models fit the data equally well for both young men and young women, and a crossvalidation conducted on an independent sample confirmed the results. CONCLUSIONS: The results provide empirical support for the inclusion of drinking driving and problem driving in an extended model of Problem Behavior Theory. Drinking-driving, drug-driving and risky driving all relate to a common construct: problem driving. These results have important implications for research and for interventions to treat or prevent drinking-driving, especially among young adults. PMID- 11925056 TI - Familial and "on-the-street" risk factors associated with alcohol use among homeless and runaway adolescents. AB - OBJECTIVE: This study investigated factors associated with alcohol use among homeless and runaway adolescents, using a risk-amplification model. METHOD: Homeless and runaway adolescents (N = 536, 60% female) were recruited and interviewed by outreach workers directly on the streets, in shelters and in drop in centers in four Midwestern states. The average age was 16 years; ages ranged from 12 to 22. RESULTS: Parent alcohol problems were indirectly linked to adolescent drinking through familial abuse and its relationship to deviant peers, time on own and risky subsistence behaviors. Parent alcohol problems also predicted offspring alcohol use through parental rejection and its association with deviant peers and with risky subsistence behaviors. The strongest direct effects on alcohol use were hanging out with antisocial friends and participating in deviant behaviors in order to survive on the street. CONCLUSIONS: This study sheds light on the nature of alcohol use in a high-risk population. Family background and "on-the-street" (time on own) factors must be taken into consideration when treating alcohol misuse in street youth. The alternative is a vicious cycle whereby homeless youth may become homeless adults. PMID- 11925057 TI - Alcohol and juvenile justice contacts: a comparison of fee-for-service and capitated medicaid mental health services. AB - OBJECTIVE: This study examines whether youth with alcohol problems are especially vulnerable to juvenile justice contact when receiving public mental health services in fee-for-service versus capitated managed-care agencies. Capitation is a payment method often accompanying managed care; a fixed fee is paid per person enrolled, for a specific time period and range of services. METHOD: Youth services were studied in a statewide, quasi-experimental design from July 1994 to June 1997; subjects were 13,365 Medicaid-eligible youth (52% female) aged 10-17. Alcohol problems were measured using a clinician-generated psychiatric diagnosis and a subjective indicator of alcohol symptoms obtained from a psychometrically tested mental health treatment intake form. Analysis methods were hazard/survival rates, log-rank tests and Cox proportional hazards models. RESULTS: As measured by the presence of symptoms, alcohol-involved youth in capitated managed care were significantly less likely to have contact with the juvenile justice system, in uncontrolled analysis. In controlled analysis, they were also less likely; however, only at the level of a trend. CONCLUSIONS: Concerns regarding capitated managed mental health care (e.g., substitution of care or "cost-shifting") found no support. There was no evidence of a shifting of alcohol-involved youth receiving mental health services under capitated managed care to the juvenile justice system. If anything, such youth saw their likelihood of juvenile justice involvement decrease. PMID- 11925058 TI - Gender differences in collegiate risk factors for heavy episodic drinking. AB - OBJECTIVE: The present research examines gender differences in the way risk factors for heavy episodic drinking operate among undergraduate students. METHOD: A web-based survey was administered to students attending a large, midwestern research university in the spring of 1999. The sample consisted of 2,041 undergraduate students with a mean age of 21.1 years; 51% were female; 72% were white, 12% Asian, 5% African American, 4% Hispanic and 7% of other races. Heavy episodic drinking was defined as having five or more drinks in a row for men and four or more for women in the last 2 weeks. Two multiple risk factor approaches, continuous and categorical, were used to examine gender differences, with the latter focusing on three drinking patterns (nonheavy episodic drinking, heavy episodic drinking, and frequent heavy episodic drinking). RESULTS: The data indicated gender differences and similarities among risk factors for heavy episodic drinking within four major domains of risk factors (background, motivational, social context and behavioral). There were significant differences in the way class year and living arrangements operated as risk factors between undergraduate men and women. Gender similarities existed for precollege drinking, drinking motivations and several behavioral measures. The categorical analysis revealed several risk factors unique to the most harmful drinking pattern. CONCLUSIONS: This study lends support for gender differences between individual risk factors for heavy episodic drinking among undergraduate students. These differences have important implications for planning research and interventions aimed at reducing collegiate heavy episodic drinking. PMID- 11925059 TI - The roles of victim and offender alcohol use in sexual assaults: results from the National Violence Against Women Survey. AB - OBJECTIVE: The roles of victim and offender alcohol use in the outcomes of sexual assault incidents (rape completion, injury and medical care) were studied. METHOD: Data from 859 female sexual assault victims identified from the National Violence against Women Survey were examined. RESULTS: Logistic regression analyses demonstrated that offender drinking was associated with greater likelihood of rape completion, but was unrelated to physical injury or medical care when victim demographics and assault characteristics were controlled. Offender aggression was the strongest predictor of both victim injury and medical care outcomes (but not rape completion). Furthermore, neither victim drinking at the time of the incident nor victim past-year drinking was significantly related to assault outcomes. CONCLUSIONS: These results suggest that offender behavior is most important for predicting assault outcomes sustained by sexual assault victims. PMID- 11925060 TI - Characteristics of mothers of children with fetal alcohol syndrome in the Western Cape Province of South Africa: a case control study. AB - OBJECTIVE: Factors associated with alcohol consumption during pregnancy and with fetal alcohol syndrome (FAS) births were examined as part of a larger epidemiologic study of FAS in a community in the Western Cape Province of South Africa. METHOD: Using retrospective case-control methodology, 31 mothers who had given birth to FAS children 6 to 9 years previously were compared with 31 matched controls on a variety of demographic, socioeconomic, drinking, family and maternity variables. Descriptive analyses were utilized to determine major differential characteristics between the two groups. RESULTS: In this community with a very high rate of FAS and rather uniform low socioeconomic status, the two groups were found to be comparable with respect to age, annual income, ethnic background, age of initiation of regular drinking, age at birth of the index child, gravidity and parity. However, mothers of FAS children reported initiating drinking at an earlier age, as well as reporting higher rates of heavy alcohol consumption in their extended family, current use of alcohol, drinking before and during pregnancy, and smoking of tobacco (percentage who smoke) during each trimester of the pregnancy. Mothers of FAS children had lower educational attainment and reported lower religiosity than control mothers. CONCLUSIONS: This study in South Africa draws upon the experience of mothers of 31 children with FAS to confirm many of the same high-risk variables identified in maternal risk studies in the United States and Europe. Some factors associated with less maternal alcohol abuse in this high-risk population were also identified, which may be helpful for implementing prevention in this region as well as in other developing countries. PMID- 11925061 TI - The effects of alcohol and provocation on aggressive behavior in men and women. AB - OBJECTIVE: The purpose of this study was to investigate the manner in which acute alcohol consumption and provocation affect aggressive behavior in men and women. METHOD: Subjects were 102 (56 men) healthy social drinkers between 21 and 35 years of age. Aggression was measured using a modified version of the Taylor Aggression Paradigm, in which electric shocks are received from and administered to a fictitious opponent during a competitive task. RESULTS: Provocation was a stronger elicitor of aggression than either gender or alcohol. Under low provocation, men were more aggressive than were women. Under high provocation, men and women were equally aggressive, except in the case of extreme aggression, in which men again exhibited higher levels than did women. Alcohol increased aggression for men but not for women. CONCLUSIONS: These results are important because there now appears to be some convergence among studies indicating that, whereas provocation and alcohol are effective in lifting inhibitions against the expression of aggression for men, only provocation seems to be an effective aggression-elicitor for women. PMID- 11925062 TI - Genetic risk for alcoholism relates to level of response to alcohol in Asian American men and women. AB - OBJECTIVE: Previous studies have shown that Asians who possess a variant aldehyde dehydrogenase allele (ALDH2*2) have lower rates of alcohol consumption and dependence. Research in Asian men has shown that those with ALDH2*2 have greater responses to alcohol than do those without this genetic variant. The present study was designed to determine whether similar levels of response to alcohol, using objective and subjective measurements, are seen in men and women with different ALDH2 genotypes. METHOD: Participants (N = 30) were 16 men and 14 women, of whom five each were heterozygous for ALDH2*2. They were evaluated in response to alcohol and placebo beverage challenges, dosed according to estimated body water. Objective and subjective responses were measured every 30 minutes from baseline to 150 minutes after ingestion. RESULTS: Men and women with ALDH2*1/*2 had greater pulse-rate increases, greater observed flushing responses and greater subjective feelings of being dizzy, drunk and high compared with ALDH2*1/*1 participants, despite having equivalent breath alcohol concentrations. ALDH2*1/*2 participants also reported being less likely to drive, following this level of intoxication, compared with ALDH2*1/*1 participants. Some gender differences were found in subjective, but not objective, responses to alcohol, with women reporting lower levels of being high, nauseated and uncomfortable and having a lower total subjective rating scale score. CONCLUSIONS: This study suggests that low risk for alcoholism based on possession of an ALDH2*2 allele relates to greater response to alcohol in both men and women. PMID- 11925063 TI - Effects of gender and family history of alcohol dependence on a behavioral task of impulsivity in healthy subjects. AB - OBJECTIVE: Substance misusers are often considered impulsive, but it is unclear whether impulsivity precedes substance misuse or develops as a consequence of it. Because alcohol dependence has a clear familial component, a study comparing impulsivity in nonaffected individuals who differ with respect to paternal history of alcohol dependence may provide evidence of familial vulnerability to impulsivity. METHOD: 122 healthy individuals participated, none of whom misused alcohol or drugs; 58 were paternal history positive (PHP) and 64 were paternal history negative (PHN) for alcohol dependence. The paternal-history groups were balanced on gender, and the four paternal-history-by-gender groups were comparable with respect to demographic features. Participants were offered choices between monetary rewards (e.g., $34) available immediately and larger rewards (e.g., $50) available after delays ranging from 1 week to 6 months. This task measures a construct of impulsivity by assessing the rates at which individuals discount rewards delayed in time. RESULTS: Although discount rates in PHP men did not differ reliably from those in PHN men, PHP women had higher discount rates than PHN women. Post hoc contrasts revealed that PHN women had lower discount rates than the other three groups. Similar results were obtained when age, education, socioeconomic status, and scores on a measure of sociopathy were used as covariates. CONCLUSIONS: Paternal history of alcohol dependence is associated with greater discount rates among women. The lack of an effect for men may suggest different mechanisms by which risk is transmitted from alcohol dependent fathers to daughters compared with sons. Further research examining these relations and the implications that delay discounting has for drinking and related behaviors is warranted. PMID- 11925064 TI - Conditioned taste aversion and alcohol drinking: strain and gender differences. AB - OBJECTIVE: The present study examined the relationship between ethanol-induced conditioned taste aversion (CTA) and ethanol oral self-administration (OSA) in male and female rats (N = 183) from three related strains not genetically selected for their ethanol preference and differing in their emotional reactivity profile. The strains used were the Wistar Kyoto (WKY), Spontaneously Hypertensive (SHR) and Wistar Kyoto Hyperactive (WKHA). We hypothesized that differences between strains in sensitivity to the aversive properties of alcohol could explain the different propensities to drink alcohol solutions. METHOD: All animals were given three conditioning trials consisting of 20-minute access to saccharin solution followed by saline or ethanol injections (0.5, 1 or 1.5 g/kg, intraperitoneally). Animals subsequently had free access to ethanol OSA for 3 weeks, followed by two CTA trials. RESULTS: Ethanol injections produce a dose dependent reduction of saccharin consumption in all animals; moreover, the strength of the CTA is gender- and strain-dependent. Taste avoidance induced by ethanol injections disturbed the initiation of ethanol OSA in two strains (WKY and WKHA) but did not change subsequent long-term ethanol consumption in either strain. In addition, voluntary alcohol drinking experience does not attenuate ethanol-induced CTA, and no association was found between ethanol-induced CTA and ethanol OSA. CONCLUSIONS: The data confirm the large variation among strains and between genders in alcohol drinking and taste-aversion learning, but suggest that there is no relationship between the sensitivity to the aversive properties of alcohol and alcohol drinking. PMID- 11925065 TI - Changes in the glucosinolate-myrosinase defense system in Brassica juncea cotyledons during seedling development. AB - Optimal defense theory (ODT) predicts that plant defenses will be allocated to plant organs and tissues in proportion to their relative fitness values and susceptibilities to attack. This study was designed to test ODT predictions on the myrosinase-glucosinolate defense system in Brassica juncea by examining the relationships between the fitness value of B. juncea cotyledons and the levels and effectiveness of cotyledon defenses. Specifically, we estimated fitness value of cotyledons during plant development by measuring plant growth and seed production after cotyledon damage or removal at successive seedling ages. Cotyledon removal within five days of emergence had a significant impact on growth and seed production, but cotyledon removal at later stages did not. Consistent with ODT, glucosinolate and myrosinase levels in cotyledons also declined with seedling age, as did relative defenses against a generalist herbivore, Spodoptera eridania, as estimated by bioassay. Declines in glucosinolates were as predicted by a passive, allometric dilution model based on cotyledon expansion. Declines in myrosinase activity were significantly more gradual than predicted by allometric dilution, suggesting active retention of myrosinase activity as young cotyledons expand. PMID- 11925066 TI - Depletion of host-derived cyanide in the gut of the eastern tent caterpillar, Malacosoma americanum. AB - Using a colorimetric procedure, we assessed the HCN-p of black cherry leaves (Prunus serotina) ingested by the eastern tent caterpillar, Malacosoma americanum, and the cyanide content of the bolus as it passed thorough the caterpillar's digestive tract and into the detritus pool. The mean HCN-p of leaves in our study area was 1902 +/- 174 (SE) ppm. Young leaves found at the tips of growing branches, which the caterpillars preferred, had a significantly higher HCN-p (3032 +/- 258 ppm) than older leaves found at the middle (1542 +/- 243 ppm) or base of the shoot (1131 +/- 159 ppm). Following a bout of overnight feeding on young leaves, the cyanide content of the foregut and midgut boluses of early sixth-instar caterpillars averaged 631 +/- 161 ppm, and 14 +/- 3 ppm, respectively, indicating that host-derived cyanide is rapidly depleted as the bolus transits the gut. Some cyanide, however, remains. In three studies, the mean cyanide content of fresh fecal pellets ranged from approximately 20 to 38 ppm, while the dried, compacted pellets ranged from 63 to 85 ppm. Food in the foreguts of mature caterpillars dispersing over the ground in search of pupation sites had 417 +/- 99 ppm cyanide. The potential impact of this egested and caterpillar-transported cyanide on the consumer and detritivore communities is discussed. PMID- 11925067 TI - Influence of flavor on goat feeding preferences. AB - Goat feeding preferences for straw pellets flavored with ryegrass (Lolium perenne, cv. Belida) or white clover (Trifolium repens, cv. Huia) aromatic extracts, obtained by means of a cold aromatic extraction method, were assessed with cafeteria trials. Prior to the trials, odor differences between the two plant species, the two aromatic extracts, and the straw pellets sprayed with the two aromatic extracts were verified using sensory analysis performed by 30 human panelists. Since odor differences observed among fresh samples were still detectable in aromatic extracts and moistened pellets, the extraction method was considered effective in reproducing plant odors. Straw pellets sprayed with either distilled water (W) or ryegrass (R) or clover (C) aromatic extracts were used to assess flavor preferences of 12 female Rossa Mediterranea goats. Sprayed pellet preference was evaluated in two sessions conducted in two consecutive weeks. Each session consisted of three two-choice presentations performed on three consecutive days. In both sessions, food intake, proportion of food intake, time spent feeding, and proportion of time spent feeding were significantly affected by pellet type (0.05 > P > 0.001). In particular, straw pellets sprayed with ryegrass extract were highly selected compared to those sprayed with clover (0.01 > P > 0.001) or water (0.01 > P > 0.001). In addition, in the second session, the clover extract was preferred to distilled water (0.05 > P > 0.01). The results of this study gave two main indications: first, goat selectivity for rye-grass against clover was consistent even when straw pellets sprayed with odors of these plants were offered, and secondly, the addition of aromatic extracts to straw pellets increased the preference for pellets. PMID- 11925068 TI - Specific attraction of fig-pollinating wasps: role of volatile compounds released by tropical figs. AB - Floral scents often act as pollinator attractants. In the case of obligate and specific plant-pollinator relationships, the role of floral signals may be crucial in allowing the encounter of the partners. About 750 Ficus species (Moraceae) are involved in such interactions, each with a distinct species of pollinating wasp (Chalcidoidea, Agaonidae). Several species have been shown to release volatile compounds, but their role in pollinator attraction has rarely been simultaneously tested. We investigated the floral scents of four tropical fig species and combined chemical analysis with biological tests of stimulation of insects. Pollinators of three species were stimulated by the odor of their associated fig species and generally not by the odor of another species. The fourth actually comprised two distinct varieties. The main compound was often a different one in each species. Floral blends of different species always shared compounds, but ratios of these compounds varied among species. PMID- 11925069 TI - Antipredator defense of biological control agent Oxyops vitiosa is mediated by plant volatiles sequestered from the host plant Melaleuca quinquenervia. AB - The weevil Oxyops vitiosa is an Australian species imported to Florida, USA, for the biological control of the invasive weed species Melaleuca quinquenervia. Larvae of this species feed on leaves of their host and produce a shiny orange secretion that covers the integument. When this secretion is applied at physiological concentrations to dog food bait, fire ant consumption and visitation are significantly reduced. Gas chromatographic analysis indicates that the larval secretion qualitatively and quantitatively resembles the terpenoid composition of the host foliage. When the combination of 10 major terpenoids from the O. vitiosa secretion was applied to dog food bait, fire ant consumption and visitation were reduced. When these 10 terpenoids were tested individually, the sesquiterpene viridiflorol was the most active component in decreasing fire ant consumption. Fire ant visitation was initially (15 min after initiation of the study) decreased for dog food bait treated with viridiflorol and the monoterpenes 1,8-cineole and alpha-terpineol. Fire ants continued to avoid the bait treated with viridiflorol at 18 microg/mg dog food for up to 6 hr after the initiation of the experiment. Moreover, ants avoided bait treated with 1.8 microg/mg for up to 3 hr. The concentrations of viridiflorol, 1,8-cineole, and alpha-terpineol in larval washes were about twice that of the host foliage, suggesting that the larvae sequester these plant-derived compounds for defense against generalist predators. PMID- 11925070 TI - Antimicrobial activity of exocrine glandular secretion of Chrysomela larvae. AB - The exocrine glandular secretions of leaf beetle larvae of the taxon Chrysomela are well-known defensive devices used against some generalist predators. Salicylaldehyde is the major repellent component of secretions emitted by larvae of Chrysomela vigintipunctata and C. lapponica, which feed on salicin-rich Salicaceae. In this study, we examined whether salicylaldehyde is also active against the entomopathogenic fungus Metarhizium anisopliae. The germination and growth of this fungus was strongly inhibited when salicylaldehyde was applied directly onto the blastospores. The salicylaldehyde concentration of the larval secretions of the tested willow feeding Chrsysomela larvae was much higher than the one necessary to display this antifungal activity. Additionally, salicylaldehyde was shown to reduce germination and growth of M. anisopliae via the gas phase over a distance of 45 mm. Further studies on the antimicrobial activity of the salicylaldehyde-containing secretions of Chrysomela larvae revealed that they act nonspecifically against prokaryotic (Escherichia coli) and eukaryotic cells (Saccharomyces cerevisiae and Trichoplusia ni). All antimicrobial and cytotoxic effects detected proved to be due to salicylaldehyde. The larval secretions of the birch-feeding allospecies of C. lapponica, that do not contain salicylaldehyde, but mainly carboxylic acids and their esters, showed no detectable effects on bacteria or fungi and no cytotoxic effects against insect cells. The results are discussed with respect to their ecological relevance. PMID- 11925072 TI - Variations in caulerpenyne contents in Caulerpa taxifolia and Caulerpa racemosa. AB - Caulerpenyne (CYN) contents was measured in two Chlorophyceae algae, Caulerpa taxifolia and Caulerpa racemosa, between July 1999 and July 2000. Sampling was performed at three stations exhibiting increasing levels of competition with the seagrass Posidonia oceanica. Significant differences were observed as a function of the Caulerpa species, the season, and the level of competition. CYN concentrations were always greater in C. taxifolia, regardless of either season or level of competition (35-80 times greater, according to the season). For a given species, maximum concentrations were recorded in autumn (September/November) and minimum values occurred in spring (April/May). CYN contents decreased with increasing level of competition, whereas frond length increased over this same gradient. It would appear that when the algae are in competition with P oceanica, Caulerpa is more inclined to accelerate vegetative growth (competition for light) than to produce secondary metabolites. PMID- 11925071 TI - Characterization and toxicity of Amanita cokeri extract. AB - The nonprotein amino acids 2-amino-3-cyclopropylbutanoic acid and 2-amino-5 chloro-4-pentenoic acid were isolated from the mushroom Amanita cokeri. The cyclopropyl amino acid is toxic to the fungus Cercospora kikuchii, the arthropod Oncopeltus fasciatus (milk weed bug), and the bacteria Agrobacterium tumefaciens, Erwinia amylovora, and Xanthomonas campestris. Toxicity to bacteria was reversible by addition of isoleucine to the medium. No toxicity was observed for 2-amino-5-chloro-4-pentenoic acid. PMID- 11925073 TI - Mycorrhizal colonization and plant growth affected by aqueous extract of Artemisia princeps var. orientalis and two phenolic compounds. AB - The effects of an aqueous extract of Artemisia princeps var. orientalis and two phenolic compounds on mycorrhizal colonization and plant growth have been investigated. Greenhouse studies showed that the inhibitory effect of the extract on mycorrhizal colonization and plant growth increased in proportion to the concentration of the extract. When the mycorrhizal test plants were treated with an increasing concentration of phenolic compounds, the mycorrhizal colonization in roots of the test plant and the plant growth were decreased. There were strong indications that mycorrhizal fungi mitigated the inhibitory influence of shoot extract of A. princeps var. orientalis and phenolic compounds. PMID- 11925074 TI - Semiochemicals and social signaling in the wild European rabbit in Australia: I. Scent profiles of chin gland secretion from the field. AB - The European rabbit (Oryctolagus cuniculus) uses the secretion of the chin gland in the maintenance of social status. Previous work has concentrated on secretion collected directly from the animal. In this study, the analysis was conducted by collecting scent marks made by free-ranging animals. Scent marks were found to be concentrated at the center of the area controlled by a social group, and at the boundaries between two adjacent social groups. Only the mark from dominant animals could be identified. Marks were also collected from the skin of rabbits, where they had been placed by the dominant individual. The mark found on the head of a subordinate animal may, in the future, be used to identify the dominant animal of the social group, who placed the mark. PMID- 11925075 TI - Cuticular hydrocarbons and novel alkenediol iacetates from wheat stem sawfly (Cephus cinctus): natural oxidation to pheromone components. AB - The cuticular lipids of the wheat stem sawfly Cephus cinctus (Hymenoptera: Cephidae) were investigated as part of a chemical ecology project with this species. The major cuticular lipids were n-alkenes and n-alkanes. Alkenes were the most abundant and exhibited dramatic sexual dimorphism. (Z)-9-Tricosene accounted for about half of the total hydrocarbon in males but was nearly absent from females. The dominant alkenes in females were (Z)-9-pentacosene and (Z)-9 heptacosene. The alkane profiles were similar in both sexes, with n-tricosane being the most abundant, followed by n-pentacosane and n-heptacosane. In both sexes, there were minor amounts of alkanes and alkenes with other chain lengths and n-alkadienes of 29 and 31 carbons. In males, about one tenth of the surface lipids consisted of (Z)-9-alkene-1, omega-diol diacetates with 22-, 24-, and 26 carbon chains. The same compounds were also detected from females but in much smaller amounts. The structures of these novel diacetates were proven by synthesis. By analogy to methyl oleate, a well-studied food lipid, the alkenes and diacetates were expected to undergo slow oxidation in air to release specific aldehydes and other volatile products, and these were generally detected in volatiles collected from living sawflies. Atmospheric oxidation of the diacetates was also demonstrated in the absence of sawflies. One product from the diacetates, 9-acetyloxynonanal, was shown in other research to be particularly active electrophysiologically and was also attractive in the field. Aldehydes from the alkenes also showed strong electrophysiological activity. The concept of volatile pheromones originating from heavy, unsaturated cuticular lipids is discussed. PMID- 11925076 TI - Pheromone components of the wheat stem sawfly: identification, electrophysiology, and field bioassay. AB - Volatiles collections and cuticular extracts of the wheat stem sawfly, Cephus cinctus Norton (Hymenoptera: Cephidae), were analyzed by coupled gas chromatographic-electroantennographic detection (GC-EAD) in order to detect possible pheromone components of this species. Volatiles collections from male and female sawflies contained the same 13 GC-EAD-active compounds that stimulated both male and female antennae. GC-EAD-active compounds were identified by mass spectrometry, and the identities were verified by acquiring or synthesizing authentic standards with identical GC retention times and mass spectra. 9 Acetyloxynonanal gave the strongest EAD response. Additional GC-EAD-active compounds included 13-acetyloxytridecanal, aldehydes with 9-16 carbon chain lengths, acids with 8-10 carbon chain lengths, and phenylacetic acid. The last was instantly detectable by human nose when several males were allowed to interact, but not from isolated insects. On a per-insect basis, grouped males overall released higher amounts of the GC-EAD-active compounds than grouped females. Notable quantitative differences between the sexes were relatively higher amounts of 9-acetyloxynonanal, phenylacetic acid, and tetradecanal from males and hexadecanal from females. In the absence of an absolute, qualitative difference between sexes, these relative differences might be important for sex recognition. Field bioassay demonstrated the attractiveness of 9-acetyloxynonanal for male and female C. cinctus. The major compounds of male and female cuticular extracts showed no GC-EAD activity, but ozonolysis of extracts gave dramatically increased amounts of GC-EAD-active material. Hence, double-bond-containing cuticular compounds are suggested as precursors of the aldehydic pheromone components. PMID- 11925077 TI - Field verification of predator attraction to minnow alarm substance. AB - Fishes such as minnows in the superorder Ostariophysi possess specialized alarm substance cells (ASC) that contain an alarm cue. Alarm substance can only be released by damage to the epidermis; thus, the release of alarm substance is a reliable indicator of predation risk. When nearby minnows detect the cue, they adopt a range of antipredator behaviors that reduce their probability of predation. Predator-predator interactions afford prey an opportunity to escape and, thus, a fitness benefit that maintains alarm substance calls over evolutionary time. Here, we present data from a simple field experiment verifying that nearby predators are attracted to minnow alarm substance because it signals an opportunity to pirate a meal. Fishing lures were baited with sponge blocks scented with either (1) water (control for sponge odor and appearance), (2) skin extract from non-ostariophysan convict cichlids (superorder Acanthopterygii, Archocentrus "Cichlasoma" nigrofasciatus) to control for general injury-released cues from fish, or (3) skin extract from fathead minnows (superorder Ostariophysi, Pimephales promelas). Predator strike frequency on each sponge type was 1, 1, and 7 for water, cichlid, and minnow cues, respectively. These data provide the first field test using fish predators of the predator-attraction hypothesis for the evolution of Ostariophysan alarm substance cells. PMID- 11925078 TI - Acquired predator recognition by fathead minnows: influence of habitat characteristics on survival. AB - In this study we conditioned fathead minnows (Pimephales promelas) to recognize the odor of a perch (Perca flavescens) by exposing them to perch odor coupled with minnow alarm cue. We then staged encounters between the predator and prey in order to assess whether the predator odor training had any effect on survival of the prey. We tested for a survival effect in the presence and absence of shelter. Our results indicate that fish trained with alarm signals to recognize predators gained a survival benefit during staged encounters with a predator and that habitat characteristics influenced the survival of conditioned fish. PMID- 11925079 TI - HHS budget for HIV/AIDS research increases 8 percent. PMID- 11925080 TI - Children, physical activity, and public health: another call to action. PMID- 11925082 TI - More on sports physical examinations. PMID- 11925081 TI - Value of preparticipation physical examination. PMID- 11925083 TI - Clinical utility of common serum rheumatologic tests. AB - Serum rheumatologic tests are generally most useful for confirming a clinically suspected diagnosis. Testing for rheumatoid factor is appropriate when rheumatoid arthritis, Sjogren's syndrome or cryoglobulinemia is suspected. Antinuclear antibody testing is highly sensitive for systemic lupus erythematosus and drug induced lupus. Anti-double-stranded DNA antibodies correlate with lupus nephritis; the titer often corresponds with disease activity in systemic lupus erythematosus. Testing for anti-Ro (anti-SS-A) or anti-La (anti-SS-B) may help confirm the diagnosis of Sjogren's syndrome or systemic lupus erythematosus; these antibodies are associated with the extraglandular manifestations of Sjogren's syndrome. Cytoplasmic antineutrophil cytoplasmic antibody testing is highly sensitive and specific for Wegener's granulomatosis. Human leukocyte antigen-B27 is frequently present in ankylosing spondylitis and Reiter's syndrome, but the background presence of this antibody in white populations limits the value of testing. An elevated erythrocyte sedimentation rate (ESR) is a diagnostic criterion for polymyalgia rheumatica and temporal arteritis; however, specificity is quite low. ESR values tend to correlate with disease activity in rheumatoid arthritis and may be useful for monitoring therapeutic response. PMID- 11925085 TI - Information from your family doctor. Chronic fatigue syndrome: how to help yourself. PMID- 11925084 TI - Chronic fatigue syndrome: evaluation and treatment. AB - Severe fatigue is a common complaint among patients. Often, the fatigue is transient or can be attributed to a definable organic illness. Some patients present with persistent and disabling fatigue, but show no abnormalities on physical examination or screening laboratory tests. In these cases, the diagnosis of chronic fatigue syndrome (CFS) should be considered. CFS is characterized by debilitating fatigue with associated myalgias, tender lymph nodes, arthralgias, chills, feverish feelings, and postexertional malaise. Diagnosis of CFS is primarily by exclusion with no definitive laboratory test or physical findings. Medical research continues to examine the many possible etiologic agents for CFS (infectious, immunologic, neurologic, and psychiatric), but the answer remains elusive. It is known that CFS is a heterogeneous disorder possibly involving an interaction of biologic systems. Similarities with fibromyalgia exist and concomitant illnesses include irritable bowel syndrome, depression, and headaches. Therefore, treatment of CFS may be variable and should be tailored to each patient. Therapy should include exercise, diet, good sleep hygiene, antidepressants, and other medications, depending on the patient's presentation. PMID- 11925086 TI - Is thrombolytic therapy effective for pulmonary embolism? AB - Pulmonary embolism is a disorder that is associated with significant morbidity and mortality. Right-sided heart failure and recurrent pulmonary embolism are the main causes of death associated with pulmonary embolism in the first two weeks after the embolic event. Thrombolysis is a potentially lifesaving therapy when used in conjunction with standard anticoagulation. However, it has significant side effects and must therefore be used with caution. Indications for thrombolysis are not well defined and are thus controversial. The only current absolute indication is massive pulmonary embolism with hypotension. Other potential indications include right heart dysfunction, recurrent pulmonary embolism and the prevention of pulmonary hypertension. However, no evidence exists to show benefit of thrombolytic therapy over standard anticoagulation therapy for recurrent pulmonary embolism, mortality or chronic complications. Bleeding is the most common complication of thrombolysis and may be fatal. PMID- 11925087 TI - Smoking cessation: integration of behavioral and drug therapies. AB - Family physicians should take advantage of each contact with smokers to encourage and support smoking cessation. Once a patient is identified as a smoker, tools are available to assess readiness for change. Using motivational interviewing techniques, the physician can help the patient move from the precontemplation stage through the contemplation stage to the preparation stage, where plans are made for the initiation of nicotine replacement and/or bupropion therapy when indicated. Continued motivational techniques and support are needed in the action stage, when the patient stops smoking. Group or individual behavioral counseling can facilitate smoking cessation and improve quit rates. Combined use of behavioral and drug therapies can dramatically improve the patient's chance of quitting smoking. A plan should be in place for recycling the patient through the appropriate stages if relapse should occur. PMID- 11925088 TI - Information from your family doctor. How to stop smoking. PMID- 11925089 TI - Osteosarcoma: a multidisciplinary approach to diagnosis and treatment. AB - The treatment of osteosarcoma requires a multidisciplinary approach involving the family physician, orthopedic oncologist, medical oncologist, radiologist and pathologist. Osteosarcoma is a mesenchymally derived, high-grade bone sarcoma. It is the third most common malignancy in children and adolescents. The most frequent sites of origin are the distal femur, proximal tibia and proximal humerus. Patients typically present with pain, swelling, localized enlargement of the extremity and, occasionally, pathologic fracture. Most patients present with localized disease. Radiographs commonly demonstrate a mixed sclerotic and lytic lesion arising in the metaphyseal region of the involved bone. Computed tomography and bone scanning are recommended to detect pulmonary and bone metastases, respectively. Before 1970, osteosarcomas were treated with amputation. Survival was poor: 80 percent of patients died from metastatic disease. With the development of induction and adjuvant chemotherapy protocols, advances in surgical techniques and improvements in radiologic staging studies, 90 to 95 percent of patients with osteosarcoma can now be treated with limb sparing resection and reconstruction. Long-term survival and cure rates have increased to between 60 and 80 percent in patients with localized disease. PMID- 11925090 TI - Information from your family doctor. Osteosarcoma. PMID- 11925091 TI - Neonatal herpes simplex virus infections. AB - Neonatal herpes simplex virus infections can result in serious morbidity and mortality. Many of the infections result from asymptomatic cervical shedding of virus after a primary episode of genital HSV in the third trimester. Antibodies to HSV-2 have been detected in approximately 20 percent of pregnant women, but only 5 percent report a history of symptomatic infection. All primary episodes of HSV and secondary episodes near term or at the time of delivery should be treated with antiviral therapy. If active HSV infection is present at the time of delivery, cesarean section should be performed. Symptomatic and asymptomatic primary genital HSV infections are associated with preterm labor and low-birth weight infants. The diagnosis of neonatal HSV can be difficult, but it should be suspected in any newborn with irritability, lethargy, fever or poor feeding at one week of age. Diagnosis is made by culturing the blood, cerebrospinal fluid, urine and fluid from eyes, nose and mucous membranes. All newborns suspected to have or who are diagnosed with HSV infection should be treated with parenteral acyclovir. PMID- 11925092 TI - Information from your family doctor. Herpes during pregnancy-what it means, what to expect. PMID- 11925093 TI - Screening for bacterial vaginosis in pregnancy: recommendations and rationale. PMID- 11925094 TI - Punch biopsy of the skin. AB - Skin biopsy is one of the most important diagnostic tests for skin disorders. Punch biopsy is considered the primary technique for obtaining diagnostic full thickness skin specimens. It requires basic general surgical and suture-tying skills and is easy to learn. The technique involves the use of a circular blade that is rotated down through the epidermis and dermis, and into the subcutaneous fat, yielding a 3- to 4-mm cylindrical core of tissue sample. Stretching the skin perpendicular to the lines of least skin tension before incision results in an elliptical-shaped wound, allowing for easier closure by a single suture. Once the specimen is obtained, caution must be used in handling it to avoid crush artifact. Punch biopsies are useful in the work-up of cutaneous neoplasms, pigmented lesions, inflammatory lesions and chronic skin disorders. Properly administered local anesthesia usually makes this a painless procedure. PMID- 11925095 TI - Information from your family doctor. Punch biopsy of the skin. PMID- 11925096 TI - AAP issues recommendations for the management of sinusitis in children. American Academy of Pediatrics. PMID- 11925097 TI - Garlic supplements and HIV medication. PMID- 11925098 TI - New study for persons with HIV. PMID- 11925099 TI - A group practice disagrees about offering contraception. PMID- 11925100 TI - Protein phosphatase 2A is involved in the regulation of protein kinase A signaling pathway during in vitro chondrogenesis. AB - We have evaluated the importance of the Ser/Thr protein phosphorylation and dephosphorylation for chondrogenesis in high-density chicken limb bud mesenchymal cell cultures (HDCs) by using H89, a cell-permeable protein kinase inhibitor, and okadaic acid (OA), a phosphoprotein phosphatase (PP)-specific inhibitor molecule. When 20 nM OA was applied to the HDCs on Days 2 and 3 of culturing, it significantly inhibited protein phosphatase 2A (PP2A), enhanced cartilage formation, and elevated the activity of cAMP-dependent protein kinase (PKA). Application of 20 microM H89 significantly decreased the activity of PKA and blocked the chondrogenesis in HDCs. Furthermore, OA enhanced cartilage formation and elevated the suppressed activity of PKA even in the H89-pretreated HDCs. cGMP dependent protein kinase was not detected in HDCs, while protein kinase Cmu (PKCmu), which is also inhibited by nanomolar concentrations of H89, was present throughout the culturing period. Neither OA nor H89 influenced the expression of the catalytic subunit of PKA or the cAMP response element binding protein, CREB. However, a significantly elevated amount of Ser-133-phosphorylated-CREB (P-CREB) was detected following addition of OA, while H89 treatment resulted in a decrease of the amount of P-CREB. Our results demonstrate that PP2A plays a role in the regulation of the PKA signaling pathway and that the phosphorylation level of CREB is influenced by the activity of both enzymes during in vitro chondrogenesis. PMID- 11925101 TI - Ligand-independent oncogenic transformation by the EGF receptor requires kinase domain catalytic activity. AB - The retroviral oncogene S3-v-erbB is a transduced, truncated form of the avian EGF (ErbB-1) receptor. Infection of avian fibroblasts with a retroviral vector expressing S3-v-ErbB results in ligand-independent cell transformation, which is accompanied by the assembly of a transformation-specific phosphoprotein signaling complex and anchorage-independent cell growth. It previously had been reported, using lysine-721 mutants (K721), that kinase domain function was required for ErbB-mediated cell transformation. However, since these initial reports, several studies using aspartate-813 mutants (D813) have demonstrated the ability of kinase-impaired ErbB receptors to induce mitogenic signal transduction pathways and cell transformation in a ligand-dependent manner. To determine the necessity of ErbB receptor kinase domain catalytic activity in ligand-independent cell transformation, we created S3-v-ErbB-K(-), a kinase-impaired oncoprotein constructed by replacing aspartate-813 with alanine (D813A). Subcellular routing as well as cell surface membrane and nuclear localization of the S3-v-ErbB-K(-) mutant receptor were unaffected by impairment of kinase activity. In contrast, avian fibroblasts expressing S3-v-ErbB-K(-) do not form the characteristic transformation-specific phosphoprotein complex, or induce soft agar colony growth in vitro. These results suggest that in contrast to ligand-dependent oncogenic signaling, ligand-independent cell transformation by a constitutively activated mutant form of the EGF receptor requires receptor kinase catalytic activity. In addition, these results demonstrate that phosphorylation and assembly of downstream signaling complexes require tyrosine phosphorylation events that are directly mediated by oncogenic forms of the EGF receptor. PMID- 11925102 TI - Desensitization of the PDGFbeta receptor by modulation of the cytoskeleton: the role of p21(Ras) and Rho family GTPases. AB - Ligand-induced PDGF-type beta receptor (PDGFbeta-R) autophosphorylation is profoundly suppressed in cells transformed by activated p21(Ras). We report here that the integrity of the actin cytoskeleton is a critical regulator of PDGFbeta R function in the presence of p21(Ras). Morphological reversion of Balb cells expressing a constitutively activated p21(Ras), with re-formation of actin stress fibers and cytoskeletal architecture, rendering them phenotypically similar to untransformed fibroblasts, allowed recovery of ligand-dependent PDGFbeta-R autophosphorylation. Conversely, disruption of the actin cytoskeleton in Balb/c 3T3 cells obliterated the normal ligand-induced phosphorylation of the PDGFbeta R. The Rho family GTPases Rac and Rho are activated by p21(Ras) and are critical mediators of cell motility and morphology via their influence on the actin cytoskeleton. Transient expression of wild-type or constitutively active mutant forms of RhoA suppressed ligand-dependent PDGFbeta-R autophosphorylation and downstream signal transduction. These studies demonstrate the necessary role of Rho in the inhibition of PDGFbeta-R autophosphorylation in cells containing activated p21(Ras) and also demonstrate the importance of cell context and the integrity of the actin cytoskeleton in the regulation of PDGFbeta-R ligand induced autophosphorylation. PMID- 11925103 TI - Normal breast epithelial cells induce apoptosis of breast cancer cells via Fas signaling. AB - Fas/Fas ligand (Fas L) death pathway is an important mediator of apoptosis. Deregulation of Fas pathway is reported to be involved in the immune escape of breast cancer and the resistance to anti-cancer drugs. In this study, we demonstrated that conditioned medium by normal breast epithelial cells (NBEC-CM) induced apoptosis of MCF-7 and T-47D Fas-sensitive cells but had no effect on MDA MB-231 Fas-resistant cells. Inhibition of PI3 kinase or NF-kappaB by specific inhibitors or transient transfections restored the sensitivity of MDA-MB-231 cells to NBEC-induced apoptosis. Moreover, the constitutive activation of NF kappaB was controlled by PI3 kinase because inhibition of PI3 kinase reduced NF kappaB activity. Inducible activation of NF-kappaB rendered MCF-7 cells resistant to NBEC-CM- and Fas agonist antibody-triggered apoptosis. Therefore, constitutive or inducible activation of PI3 kinase and/or NF-kappaB in breast cancer cells rendered them resistant to NBEC-triggered apoptosis. In addition, Fas neutralizing antibody and dominant negative Fas abolished NBEC-triggered apoptosis. Western blot and confocal microscopy analysis showed an increase of membrane Fas/Fas L when cells were induced into apoptotis by NBEC-CM. Taken together, these data show that NBEC induced apoptosis in breast cancer cells via Fas signaling. PMID- 11925104 TI - CRM1-mediated nuclear export determines the cytoplasmic localization of the antiapoptotic protein Survivin. AB - Survivin is a member of the inhibitor of apoptosis (IAP) family of negative regulators of programmed cell death that is frequently overexpressed in human tumors. Survivin is not only involved in the regulation of apoptosis, but is also known to play a role in the control of cell cycle progression at the G2/M phase. Survivin is a predominantly cytoplasmic protein expressed in a cell cycle dependent manner, but the mechanism(s) that determine its nuclear-cytoplasmic localization have not been described. In this study, we report that Survivin is a nuclear shuttling protein that is actively exported from the nucleus via the CRM1 dependent pathway. Nuclear export of Survivin is independent of the export of other shuttling proteins that control the G2/M phase transition, such as cyclin B1 and cdc25. The carboxy-terminal domain of Survivin is both necessary and sufficient for its nuclear export, although this region does not contain a functional leucine-rich nuclear export signal. Differences in the amino acid sequence of this region determine the dramatically different localization of Survivin (in the cytoplasm) and its splicing variant Survivin-DeltaEx3 (in the nucleus). The carboxy-terminal end of Survivin-DeltaEx3 contains a bipartite nuclear localization signal, not present in Survivin, which mediates its strong nuclear accumulation. These data suggest that active transport between the nucleus and cytoplasm may constitute an important regulatory mechanism for Survivin function. PMID- 11925105 TI - Eukaryotic cell locomotion depends on the propagation of self-organized reaction diffusion waves and oscillations of actin filament assembly. AB - Actin filament (F-actin) assembly kinetics determines the locomotion and shape of crawling eukaryotic cells, but the nature of these kinetics and their determining reactions are unclear. Live BHK21 fibroblasts, mouse melanoma cells, and Dictyostelium amoebae, locomoting on glass and expressing Green Fluorescent Protein-actin fusion proteins, were examined by confocal microscopy. The cells demonstrated three-dimensional bands of F-actin, which propagated throughout the cytoplasm at rates usually ranging between 2 and 5 microm/min in each cell type and produced lamellipodia or pseudopodia at the cell boundary. F-actin's dynamic behavior and supramolecular spatial patterns resembled in detail self-organized chemical waves in dissipative, physico-chemical systems. On this basis, the present observations provide the first evidence of self-organized, and probably autocatalytic, chemical reaction-diffusion waves of reversible actin filament assembly in vertebrate cells and a comprehensive record of wave and locomotory dynamics in vegetative-stage Dictyostelium cells. The intensity and frequency of F-actin wavefronts determine locomotory cell projections and the rotating oscillatory waves, which structure the cell surface. F-actin assembly waves thus provide a fundamental, deterministic, and nonlinear mechanism of cell locomotion and shape, which complements mechanisms based exclusively on stochastic molecular reaction kinetics. PMID- 11925106 TI - Gap junctions assemble in the presence of cytoskeletal inhibitors, but enhanced assembly requires microtubules. AB - The role of cytoskeletal elements in gap junction (GJ) assembly has been studied using Novikoff hepatoma cells treated with cytochalasin B (CB) to disrupt actin filaments or with colchicine or nocodazole to disrupt microtubules. After 60 min of cell reaggregation, freeze-fracture was used to evaluate quantitatively the "initiation," "maturation," and "growth" phases of GJ assembly. The development of junctional permeability to fluorescent dyes was also analyzed. The only effects of CB on the structure or permeability of the developing junctions involved an elongation of GJ aggregates and a small decrease in formation plaque areas. Colchicine (but not the inactive form, lumicolchicine) prevented the enhancement of GJ growth by cholesterol, but its effect on basal growth was equivocal. Nocodazole inhibited the growth of GJ, even under basal conditions, without an effect on initiation. Nocodazole also blocked the forskolin-enhanced increase in the growth of GJs and, in living MDCK cells, reduced the movement of transport intermediates containing green fluorescent protein-tagged connexin43. Thus, neither actin filaments nor microtubules appear to restrict GJ assembly by anchoring intramembrane GJ proteins, nor are they absolutely required for functional GJs to form. However, microtubules are necessary for enhanced GJ growth and likely for facilitating connexin trafficking under basal conditions. PMID- 11925107 TI - Mutational analysis of the central centromere targeting domain of human centromere protein C, (CENP-C). AB - Human centromere protein C (CENP-C) is an essential component of the inner kinetochore plate. A central region of CENP-C can bind DNA in vitro and is sufficient for targeting the protein to centromeres in vivo, raising the possibility that this domain mediates centromere localization via direct DNA binding. We performed a detailed molecular dissection of this domain to understand the mechanism by which CENP-C assembles at centromeres. By a combination of PCR mutagenesis and transient expression of GFP-tagged proteins in HeLa cells, we identified mutations that disrupt centromere localization of CENP C in vivo. These cluster in a 12 amino acid region adjacent to the core domain required for in vitro DNA binding. This region is conserved between human and mouse, but is divergent or absent in invertebrate and plant CENP-C homologues. We suggest that these 12 amino acids are essential to confer specificity to DNA binding by CENP-C in vivo, or to mediate interaction with another as yet unidentified centromere component. A differential yeast two-hybrid screen failed to identify interactions specific to this sequence, but nonetheless identified 14 candidate proteins that interact with the central region of CENP-C. This collection of mutations and interacting proteins comprise a useful resource for further elucidating centromere assembly. PMID- 11925108 TI - Testis fascin (FSCN3): a novel paralog of the actin-bundling protein fascin expressed specifically in the elongate spermatid head. AB - During spermiogenesis, significant morphological changes occur as round spermatids are remodeled into the fusiform shape of mature spermatozoa. These changes are correlated with a reorganization of microfilaments and microtubules in the head and tail regions of elongating spermatids. There is also altered expression of specialized actin- and tubulin-associated proteins. We report the characterization of a novel, spermatid-specific murine paralog of the actin bundling protein fascin (FSCN1); this paralog is designated testis fascin or FSCN3. Testis fascin is distantly related to fascins but retains its primary sequence organization. cDNA clones of mouse testis fascin predict a 498 amino acid protein of molecular mass 56 kD that shares 29% identity with mouse fascin. Mapping of murine and human FSCN3 genes shows localization to the 7q31.3 chromosome. Northern analysis indicates that FSCN3 expression is highly specific to testis and that in situ hybridization further restricts expression to elongating spermatids. Antibodies raised against recombinant FSCN3 protein identify a band at 56 kD in testis, epididymis, and epididymal spermatozoa, suggesting that testis fascin persists in mature spermatozoa. In accord with the in situ hybridization results, immunofluorescent microscopy localizes testis fascin protein to areas of the anterior spermatid head that match known distributions of F-actin in the dorsal and ventral subacrosomal spaces. It is possible that testis fascin may function in the terminal elongation of the spermatid head and in microfilament rearrangements that accompany fertilization. PMID- 11925109 TI - Stimulation of matrix metalloproteinase-9 expression in human fibrosarcoma cells by synthetic matrix metalloproteinase inhibitors. AB - Enhanced expression and activation of matrix metalloproteinase-2 (MMP-2) and MMP 9 have been associated with tumor progression, invasion, and metastasis. The use of synthetic MMP inhibitors to block the proteolytic activity of these enzymes recently emerged as a potential therapeutic tool to treat cancer. In this study, we report that GI129471, a synthetic broad-spectrum MMP inhibitor, efficiently reduced the in vitro invasiveness of HT1080 cells through type IV collagen, a major component of basement membranes. This reduced invasion was paralleled by a complete inhibition of pro-MMP-2 activation; however, GI129471 strongly increased the amount of secreted pro-MMP-9, which could be subsequently activated through a plasminogen-dependent mechanism. Quantitative RT-PCR and northern blot analysis revealed that GI129471 specifically increased the MMP-9 mRNA steady-state level. Moreover, transient transfection of HT1080 cells with beta-galactosidase reporter vectors containing different lengths of the 5'-flanking region of the MMP-9 gene revealed an upregulation of the transcriptional activity of the corresponding promoter. Well-known modulators of MMP-9 expression such as Il-1beta and TNF alpha were not involved in this upregulation. These findings emphasize the complexity of the regulation of MMP expression and the requirement for a detailed characterization of the potential adverse side effects associated with the use of broad-spectrum MMPIs. PMID- 11925110 TI - Expression of different mutant p53 transgenes in neuroblastoma cells leads to different cellular responses to genotoxic agents. AB - The involvement of p53 as a determinant of chemosensitivity or radiosensitivity is not well understood and is complicated by numerous contradictory reports. Here we have addressed this issue using a series of isogenic clones derived from two neuroblastoma cell lines that express wild-type p53 genes, Nub7 and IMR32. Two different mutant p53 transgenes were used in an attempt to disrupt p53 function in the clones. Our findings indicate that the cellular response is dependent on the genotoxic agent used as well as on the specific p53 transgene used. Cellular radiosensitivity showed no association with apoptosis or with the ability of the cells to arrest in G1 after irradiation. An association was observed, however, between gamma-radiation sensitivity and DNA double-strand break rejoining activity. PMID- 11925111 TI - Bone morphogenetic protein-7 (OP1) and transforming growth factor-beta1 modulate 1,25(OH)2-vitamin D3-induced differentiation of human osteoblasts. AB - Bone morphogenetic proteins (BMPs) and transforming growth factor-beta (TGFbeta) are potent regulators of osteoblast differentiation and proliferation, processes that are crucial in bone remodeling. BMPs and TGFbeta act in concert with other local factors and hormones, among them 1,25(OH)2-vitamin D3 and insulin. Here we show that BMP7 inhibits 1,25(OH)2-vitamin D3-induced differentiation of human osteoblasts, whereas TGFbeta1 stimulates it, as assessed by assays for alkaline phosphatase (ALP) induction, matrix mineralization, and morphology changes. BMP7 or TGFbeta1 alone affects the differentiation of human osteoblasts. Similar results were obtained in assays for ALP induction using conditionally immortalized human osteoblasts (hFOB) and primary osteoblasts obtained from trabecular bone of the femoral head after hip replacement surgery. BMP7 stimulation led to a decrease of 1,25(OH)2-vitamin D3-induced binding of nuclear proteins to a vitamin D response element, as shown by electrophoretic mobility shift assay. Our results suggest that 1,25(OH)2-vitamin D3 modulates in opposite ways the effects of BMP7 and TGFbeta1 on osteoblast differentiation. PMID- 11925112 TI - Screening for ovarian cancer-risk, education, worry: path to appropriate use? PMID- 11925113 TI - Worry about ovarian cancer risk and use of ovarian cancer screening by women at risk for ovarian cancer. AB - OBJECTIVE: This study examined reports of perceived risk of ovarian cancer, worry, and screening use in a large sample of women. While screening for asymptomatic women is not generally recommended, in 1994 a consensus conference concluded that women with multiple affected relatives are at high risk for ovarian cancer and should be encouraged to participate in screening. The consensus report also suggested that women with a single affected first-degree relative are at elevated risk and while these women were not encouraged to get screening it was suggested that they may choose to pursue screening outside of a randomized trial [NIH Consensus Conference. JAMA 1995;273(6) 491-7]. METHODS: A total of 3257 women participated in this research by completing a mailed survey on ovarian cancer risk, worry, and use of screening. One hundred forty-two of these women were at high risk for this disease due to a strong family history. An additional 144 women were at elevated risk due to a single first-degree affected relative with ovarian cancer. RESULTS: Family history did predict perceived risk, difficulties due to worry, and use of ovarian cancer screening. However, the group of women most likely to report high levels of perceived risk and to have received screening for ovarian cancer were women with a single affected relative rather than those at high risk, for whom screening is recommended. CONCLUSIONS: These results suggest that many women need additional education about ovarian cancer risk. Most women overestimated their risk for this disease. Some average risk women get screening although it is not recommended outside of randomized trials, and a significant percentage of women at high risk fail to get recommended screening. PMID- 11925114 TI - Hereditary ovarian cancer. Lessons from the first twenty years of the Gilda Radner Familial Ovarian Cancer Registry. PMID- 11925115 TI - Anthracyclines in the treatment of gynecologic malignancies. AB - OBJECTIVE: The purpose of this article is to present a summary of the pharmacology of anthracyclines as well as to review the results of clinical trials including patients with gynecologic malignancies treated with anthracycline-based therapy. METHODS: We performed a MEDLINE literature search of relevant clinical trials for the scope of this review that evaluated anthracycline-based therapy in gynecologic malignancies. RESULTS: Doxorubicin has established activity in carcinomas that arise in the ovary, uterine cervix, and endometrium as well as in uterine sarcomas. However, doxorubicin has structural characteristics that limit its efficacy and safety. Newer anthracyclines with distinct structure, pharmacokinetics, pharmacodynamics, and toxicity profiles have been developed to overcome the limitations of doxorubicin and to further exploit the activity of anthracyclines. Epirubicin is characterized by a structural formula that confers similar cytotoxic antitumor activity with fewer associated side effects than its analogue. Most recently, pegylated liposomal formulations, with distinct pharmacokinetic properties and a favorable toxicity profile, have shown antitumor activity as salvage therapy in ovarian cancer. Intraperitoneal mitoxantrone is also associated with activity in ovarian cancer; however, its clinical use is limited by the severity of local adverse effects. CONCLUSIONS: The role of anthracyclines in the management of advanced gynecologic malignancies is important as part of first-line therapy or as a salvage approach. Newer anthracycline agents such as epirubicin and pegylated liposomal doxorubicin are associated with a more favorable toxicity profile. Clinical trials are under way to further explore the role of newer anthracycline-based regimens as first line or salvage treatment in gynecologic malignancies. PMID- 11925116 TI - Serum vascular endothelial growth factor and serum leptin in patients with cervical cancer. AB - OBJECTIVE: Serum levels of vascular endothelial growth factor (VEGF) can be seen as surrogate markers of angiogenesis. Recently, leptin, which is involved in the control of satiety and energy expenditure, was also shown to modulate angiogenesis. As angiogenesis plays an abundant role in cervical carcinogenesis, we evaluated serum VEGF and leptin in patients with cervical intraepithelial neoplasia (CIN) and cervical cancer. METHODS: Serum VEGF and leptin were measured in 84 patients with cervical cancer, in 28 patients with CIN I-III, and in 35 healthy women, using a commercially available enzyme-linked immunosorbent assay and radioimmunoassay, respectively. RESULTS: Serum VEGF was significantly elevated in patients with cervical cancer and in patients with CIN I-III compared to healthy women. In patients with cervical cancer serum VEGF was significantly correlated with tumor stage, but not with lymph node involvement and histological grade. Univariate and multivariate analyses showed that elevated pretreatment serum VEGF was not associated with the duration of disease-free and overall survival. Serum leptin did not differ among patients with cervical cancer, patients with CIN I-III, and healthy women. Serum leptin was significantly correlated with body mass index (BMI). All further analyses were performed with absolute and serum leptin corrected by BMI. No differences in serum leptin could be ascertained between patients with cervical cancer and patients with CIN I-III. Serum leptin was not associated with any clinicopathological parameter and patients' survival. No correlation between serum VEGF and leptin was found. CONCLUSIONS: It can be speculated that serum VEGF might be used as a surrogate marker of angiogenesis in patients with cervical cancer. Our data support the concept that VEGF plays a role in malignant transformation and tumor growth, but not in the lymphatic spread of cervical cancer. This is the first report on leptin in a gynecological malignancy. Our results show that serum leptin falls short of being a useful marker in patients with cervical cancer. PMID- 11925117 TI - Is lack of response to single-agent chemotherapy in gestational trophoblastic disease associated with dose scheduling or chemotherapy resistance? AB - OBJECTIVE: The aim of this study was to determine whether in the management of low-risk gestational trophoblastic neoplasia (GTN) the administration of 5-day courses of 12 microg/kg actinomycin D is effective following the failure of 1.25 mg/m(2) "pulsed" actinomycin D. METHODS: Patients with low-risk GTN who failed to respond to 1.25 mg/m(2) pulsed actinomycin were switched to the 5-day course of 12 microg/kg actinomycin. RESULTS: Patients with low-risk GTN who failed to respond to pulsed actinomycin were changed to the same chemotherapy agent, actinomycin D, given as a 5-day course at 12 microg/kg. Four of the five responded and one required methotrexate to achieve remission. CONCLUSIONS: Pulsed biweekly actinomycin and pulsed weekly methotrexate have been shown to have a higher failure rate than the 5-day regimens of the same medications. This study demonstrates that failure of pulsed actinomycin may be successfully treated by a 5-day course of the same medication. It appears that with the pulsed regimens cytotoxic exposure of trophoblastic cells to the medication is too brief and the 5-day course permits more cells to be in cycle. It is suggested that, following failure of a pulsed regimen, the patient is given the same chemotherapy as a 5 day course, rather than switching from actinomycin to methotrexate or vice versa. This conserves options for chemotherapy in GTN. PMID- 11925118 TI - Combination antigene therapy targeting c-myc and c-erbB(2) in the ovarian cancer COC(1) cell line. AB - OBJECTIVE: Antigene therapy targeting only one oncogene in ovarian cancer has made much progress, although it still has some limitations. To explore the potential for combination antigene therapy in ovarian cancer, we examined the in vitro effects of liposmal antisense phosphorothioate oligodeoxynucleotides targeting c-erbB(2) and c-myc (LF-c-erbB(2)/c-myc AS-ODNs) in the human ovarian cancer COC(1) cell line. METHODS: COC(1) cells were treated differently as follows: group A with single LF-c-erbB(2) AS-ODNs; group B with single LF-c-myc AS-ODNs; group C with combination LF-c-erbB(2)/c-myc AS-ODNs; and group D as untreated control. Cell proliferation was studied by MTT assay and clonal cultures. RT-PCR was used to measure gene expression of c-erbB(2) and c-myc before and after transfection. Morphologic changes in the COC(1) cells were observed with the electron microscope. RESULTS: Single antigene therapy targeting c-erbB(2) or c-myc could reduce target gene expression and inhibit COC(1) cell growth by 61.9 +/- 9.3 and 64.5 +/- 11.2%, respectively. However, combination antigene therapy could not only suppress expression of c-erbB(2) and c-myc simultaneously, but also inhibit COC(1) cell proliferation with a higher inhibitory rate of 82.6 +/- 12.1%. Apart from that, the combination agents could induce COC(1) cell apoptosis. CONCLUSIONS: Our study suggests that combination antigene therapy targeting c-erbB(2) and c-myc can inhibit COC(1) cell proliferation and gene expression of c-erbB(2) and c-myc. Furthermore, its effectiveness is much higher than that of individual antigene therapy. PMID- 11925119 TI - Effects of a lytic peptide conjugated to beta HCG on ovarian cancer: studies in vitro and in vivo. AB - OBJECTIVE: The aim of this study was to determine the in vitro and in vivo effects of the lytic peptide, hecate, alone and conjugated to a 15-amino-acid fragment of the beta-chain of hCG (hecate-beta hCG) on the ovarian carcinoma cell line NIH: OVCAR-3 and determine the expression of luteinizing hormone (LH)/human chorionic gonadotropin (hCG) receptors in cell cultures and tumor tissues. METHODS: For in vitro studies, hecate or hecate-beta hCG was added to cultures of ovarian cancer cells in the presence or absence of estradiol or follicle stimulating hormone. The cytotoxicity of lytic peptides was measured by trypan blue exclusion and lactate dehydrogenase release. For in vivo studies, OVCAR-3 xenografts were established in female athymic nude mice which were then treated once per week for 3 weeks with hecate or hecate-beta hCG via the lateral tail vein. An immunohistochemical method was used to analyze the expression of LH/hCG receptor in tumor and culture cells. RESULTS: In in vitro studies, both hecate beta hCG and hecate destroyed ovarian cancer cells (NIH: OVCAR-3) in a dose dependent manner. Removal of steroids from the culture medium reduced the sensitivity of the OVCAR-3 cell line to the hecate-beta hCG in a reversible manner. In in vivo studies, the average tumor volume and tumor burden in lytic peptide treated animals were reduced. In the groups of animals treated by hecate, hecate-beta hCG, and estradiol + hecate-beta hCG, tumor volumes after treatment expressed as a percentage of increase (197.4 +/- 21.72, 199.0 +/- 18.57, and 193.8 +/- 22.94%, respectively) were reduced, compared to control (263.0 +/- 21.72%) animals (P < 0.05). Immunocytochemical studies revealed the expression of LH/hCG receptor protein in the OVCAR-3 cells and tumor tissues. CONCLUSION: Hecate-beta hCG is a putative candidate for treating ovarian cancer. PMID- 11925120 TI - Positron emission tomography/computed tomography imaging for the detection of recurrent ovarian and fallopian tube carcinoma: a retrospective review. AB - PURPOSE: Imaging modalities to evaluate ovarian/fallopian tube cancer patients for recurrence are limited. Positron emission tomography (PET), computed tomography (CT), magnetic resonance imaging (MRI), and ultrasound lack the sensitivity to consistently detect recurrence or measurable disease in these patients. A new technique combines PET and CT (PET/CT) images to identify increased metabolic activity and to locate that signal with improved anatomic specificity. The objective of this study is to compare PET/CT, CT, and histologic findings in patients with recurrent ovarian/fallopian tube cancers. METHODS: Retrospective chart review of eight patients with primary ovarian (n = 6) or fallopian tube (n = 2) cancer was performed. All eight patients underwent initial cytoreductive surgery. Five patients initially received chemotherapy, one received radioactive phosphorus ((32)P), one received tamoxifen, and one received no therapy. Seven of eight patients had a suspected recurrence based on clinical examination, elevated CA-125 level, and/or abnormal CT findings; one patient requested a PET/CT. Histologic findings from surgery were correlated with PET/CT and CT findings. RESULTS: All eight patients had positive histology, and of these, seven patients had a negative CT and five patients had lesions that were correctly identified by PET/CT. CONCLUSIONS: Five of the eight (62%) patients had recurrent disease based on correlative histology with a positive PET/CT and a negative CT. These preliminary findings suggest that combined PET/CT may be an effective means of identifying patients with recurrent ovarian/fallopian tube cancer. Such patients could potentially proceed to salvage treatment and avoid the morbidity and expense of surgical assessment. Pilot studies comparing CT, PET, PET/CT, and histologic findings are underway. PMID- 11925121 TI - Evidence of an association between human papillomavirus and impaired chemotherapy induced apoptosis in cervical cancer cells. AB - OBJECTIVES: The aim of this study was to determine cervical cancer cell sensitivity to chemotherapy-induced apoptosis based on human papillomavirus (HPV) status. METHODS: CaSki (HPV-positive) and C33A (HPV-negative) cells were treated with camptothecin or cisplatin. Cellular viability was determined by trypan blue exclusion. Apoptotic indexes were determined by flow cytometric analysis of annexin V labeling and morphological changes. Mitochondrial release of cytochrome c was determined by immunofluorescence using confocal microscopy. Caspase 3 activation and bax expression were assessed by immunoblotting. RESULTS: CaSki cells displayed chemoresistance to both camptothecin and cisplatin. Low response to apoptogenic stimuli was evidenced by a marginal increase in the apoptotic cell fraction after camptothecin treatment (22.9 +/- 2.56%) compared with control (17.8 +/- 1.95%). Cisplatin (14.8 +/- 1.01%) caused a slight decrease in apoptosis. Mitochondrial release of cytochrome c and cleavage of caspase 3 could not be demonstrated in CaSki cells after treatment. Despite p53 mutation, C33A cells were sensitive to the antiproliferative effects of camptothecin and cisplatin. Mean apoptotic events were 17.5 +/- 0.33 for control, 42 +/- 1.76 for cisplatin, and 38.1 +/- 0.75 for camptothecin. An intact cytochrome c pathway was demonstrated in C33A cells leading to cleavage of caspase 3 after camptothecin treatment. The constitutive bax expression demonstrated in both cell lines displayed no change after camptothecin treatment. CONCLUSION: HPV-positive cervical cancer cells have an inherent resistance to chemotherapy-induced apoptosis. HPV-dependant inactivation of apoptotic regulators such as p53 and blockage of downstream events such as cytochrome c release and caspase 3 activation might be elemental to this cellular survival advantage provided by high-risk oncogenic papillomavirus. PMID- 11925122 TI - The effects of 5% imiquimod cream on high-grade vulval intraepithelial neoplasia. AB - OBJECTIVES: The aim of this study was to investigate the effects of topical 5% Imiquimod (3M Pharmaceuticals, St. Paul, Minnessota) on high-grade vulval intraepithelial neoplasia (VIN). METHODS: A prospective uncontrolled observational study was performed. Fifteen patients with histologically confirmed VIN 3 were asked to self-administer 5% Imiquimod cream to their vulval lesions up to three times weekly for 16 weeks. Review was conducted at 1, 2, 3, 4, 6, and 9 months postrecruitment. Lesions were photodocumented and at 4 months any areas demonstrating a clinical response were biopsied. RESULTS: Of 15 patients recruited, 4 demonstrated a clinical improvement in their disease, 3 of whom had negative biopsies posttreatment. Local side effects limited the frequency of application such that 7 patients applied the cream once weekly, 6 twice weekly, and 2 three times weekly. CONCLUSIONS: 5% Imiquimod cream appears to have an effect when used on high-grade VIN. The frequency of application was limited by local side effects which may have reduced the clinical responses seen. Measures to alleviate local side effects may allow more aggressive use of Imiquimod and lead to improved responses. PMID- 11925123 TI - Systematic review of first-line chemotherapy for newly diagnosed postoperative patients with stage II, III, or IV epithelial ovarian cancer. AB - OBJECTIVE: The aim of this study was to determine the optimal postoperative chemotherapy regimen for women with newly diagnosed stage II, III (micro or macro), or IV epithelial ovarian cancer. METHODS: A systematic search was conducted to find randomized controlled trials and meta-analyses published between 1980 and April 2001. RESULTS: A published meta-analysis found that, compared with non-platinum-based regimens, platinum, alone or in combination with other agents, improved survival when used as first-line chemotherapy for ovarian cancer. The meta-analysis did not detect a difference in efficacy between cisplatin and carboplatin. A published randomized trial of cisplatin plus paclitaxel versus carboplatin plus paclitaxel did not detect a significant difference in survival between these regimens. In two randomized trials, treatment with paclitaxel plus cisplatin resulted in improved survival compared with cyclophosphamide plus cisplatin. A randomized trial of paclitaxel plus cisplatin versus paclitaxel alone versus cisplatin alone detected no differences in survival among the three treatment groups. While hematologic adverse effects were more frequent with carboplatin than with cisplatin, nonhematologic adverse effects were less frequent with carboplatin. The addition of paclitaxel to cisplatin did not appear to increase the incidence of serious adverse effects. CONCLUSIONS: Intravenous carboplatin plus paclitaxel is the recommended postoperative chemotherapy regimen for newly diagnosed stage II-IV epithelial ovarian cancer. Intravenous cisplatin plus paclitaxel may also be considered a treatment option. Intravenous carboplatin as a single agent may be considered a treatment option in patients for whom paclitaxel is contraindicated or in patients who are unwilling to accept the adverse effects of paclitaxel chemotherapy. PMID- 11925124 TI - Laparoscopic retroperitoneal lymphadenectomy followed by immediate laparotomy in women with cervical cancer: a gynecologic oncology group study. AB - OBJECTIVES: From May 1993 until June 1997, the Gynecologic Oncology Group undertook a study of women with cancer of the cervix (Stage IA, IB, and IIA) who were about to undergo radical abdominal hysterectomy, pelvic lymphadenectomy, and aortic lymph node sampling. Immediately before this surgery, laparoscopy was performed and removal of the lymph nodes was attempted. There were two objectives: (1) to obtain information on the adverse effects and difficulties associated with laparoscopic lymph node removal and (2) to determine the adequacy of the lymph node removal. METHODS: Four methods determined the adequacy of the lymph node removal: (1) the surgeon's opinion during laparoscopy, (2) a photographic record reviewed by two independent observers, (3) inspection of the surgical sites at laparotomy, and (4) lymph node count. RESULTS: Seventy-three women were entered onto the study. Four patients were judged ineligible; 2 did not undergo laparoscopy and 17 women did not complete laparoscopic surgery because of metastatic lymph nodes judged unresectable or complications. Ten women were inevaluable. The remaining 40 women were completely evaluable for protocol objectives. All cases of bilateral laparoscopic aortic lymph node sampling were judged adequate by all four methods of evaluation. For laparoscopic pelvic lymphadenectomy 6 were judged incomplete at laparotomy, 3 of which were judged incomplete by independent reviewers. The mean number of right pelvic nodes removed was 16.6; left pelvic nodes 15.5; right aortic nodes 6.2; and left aortic nodes 5.9. CONCLUSIONS: Laparoscopic bilateral aortic lymph node sampling appeared to be reasonably safe and feasible. Laparoscopic therapeutic bilateral pelvic lymphadenectomy, although having a reasonable complication rate, demonstrated problems regarding adequacy, which are probably correctable. PMID- 11925125 TI - Phase II trial of topotecan and cisplatin in persistent or recurrent squamous and nonsquamous carcinomas of the cervix. AB - OBJECTIVE: Cisplatin is a standard treatment in advanced, recurrent cervical cancer. Because topotecan is an established treatment in gynecologic malignancies such as ovarian cancer and exhibits nonoverlapping toxicity with cisplatin, a phase II trial was conducted to evaluate the tolerability and antitumor activity of a cisplatin/topotecan doublet in persistent or recurrent cervical cancer patients. METHODS: Patients with bidimensionally measurable persistent or recurrent squamous cell and non squamous cell cervical cancer and adequate bone marrow were enrolled. Patients received 50 mg/m(2) of cisplatin intravenously over 1 h on Day 1 and 0.75 mg/m(2) of topotecan intravenously over 30 min on Days 1, 2, and 3 of 21-day cycles for six cycles or until disease progression. Tumor response and regimen toxicity were assessed using established Gynecologic Oncology Group criteria. RESULTS: Thirty-two of 35 enrolled patients were evaluable for toxicity and tumor response. All but 2 evaluable patients had received previous radiotherapy. No patient received prior chemotherapy. The cisplatin/topotecan doublet was well tolerated, with 77 and 78% of courses given without interruption or delay and at full doses, respectively. As anticipated, the most common toxicity was hematologic, with grade 3/4 neutropenia and thrombocytopenia reported in 30 and 10% of cycles, respectively. The overall response rate was 28% (9/32), with 3 complete and 6 partial responses. The antitumor response in nonirradiated fields (30%) was similar to the response observed in previously irradiated fields (33%), suggesting good drug penetration. Median duration of response was 5 months (range, 2 to 15+ months). An additional 9 (28%) patients achieved stable disease. Median survival was 10 months, with 3 patients in lasting remission. CONCLUSIONS: These results demonstrate that the cisplatin/topotecan combination is safe, well tolerated, and active in persistent or recurrent cervical cancer patients. A phase III, multicenter trial is under way (cisplatin/topotecan versus cisplatin) based on these favorable results to confirm the safety and efficacy profile in this patient population. PMID- 11925126 TI - Antiproliferative and apoptotic effects of iron chelators on human cervical carcinoma cells. AB - OBJECTIVE: Cervical carcinoma is a human papillomavirus (HPV)-associated cancer for which treatment options still mainly rely on surgical procedures, with or without adjuvant radiotherapy and chemotherapy. As iron may participate in the pathogenesis of viral infections and cancer in several ways, the present study was designed to investigate the effect of iron chelation on HPV-16- and HPV-18 positive cervical carcinoma cell lines. METHODS: Desferrioxamine and deferiprone, two chemically unrelated iron chelators, were used to investigate the effect of iron chelation on SiHa and HeLa cells. Proliferation was investigated by cells counts, by [(3)H]thymidine uptake assay, and by immunostaining with Ki-67 and proliferating cell nuclear antigen (PCNA). Apoptosis was determined by morphological analysis, by a TUNEL assay, and by flow cytometry detecting FITC conjugated annexin-V. RESULTS: Desferrioxamine and deferiprone induced a time- and dose-dependent inhibition of SiHa and HeLa cell growth. The inhibition of cell growth was associated with a decrease in the expression of both stable and total PCNA and Ki-67, a proliferation marker whose expression may predict survival in uterine cervical carcinoma. TUNEL assay, flow cytometry with annexin V-fluorescein, and morphological analysis indicated that iron chelation also induced a time- and dose-dependent apoptosis of both cell lines. This apoptotic effect was prevented by the addition of exogenous iron. CONCLUSION: These results show that iron chelation inhibits the growth and induces the apoptosis of HPV positive carcinoma cells. This suggests that iron chelators may represent a potential therapeutic approach for the management of cervical carcinoma. PMID- 11925127 TI - Anti-invasive effect of MMI-166, a new selective matrix metalloproteinase inhibitor, in cervical carcinoma cell lines. AB - OBJECTIVES: The aim of our study was to evaluate the anti-invasive effect of MMI 166, a new matrix metalloproteinase (MMP) inhibitor in cervical carcinoma cell lines. METHODS: We analyzed the invasive activities of cervical carcinoma cell lines (CAC-1, CaSki, and SiHa) and the gene expression of various matrix proteinases (matrix metalloproteinase-1 [MMP-1], MMP-2, MMP-9, membrane-type MMP type 1 [MT1-MMP], MT2-MMP, and MT3-MMP) and their inhibitors (tissue inhibitor of metalloproteinase type 1 [TIMP-1] and TIMP-2). The effect of MMI-166 was analyzed by in vitro invasion assay. The cytotoxicity of MMI-166 was determined by MTT assay. The gelatinase activity was analyzed by gelatin zymography. RESULTS: Cervical carcinoma cell lines, which produced both MMP-2 and MT1-MMP, showed invasive capacity in the in vitro invasion assay. The invasion of cervical carcinoma cells was suppressed by MMI-166. No remarkable suppression of the proliferation by MMI-166 was observed in the MTT assay. Gelatin zymography revealed complete suppression of MMP-2 activity by MMI-166. CONCLUSIONS: MMI-166 inhibited the MMP-2 activity in cervical carcinoma cells and it is useful for the regulation of cervical carcinoma cell invasion. PMID- 11925128 TI - Serum, pleural effusion, and ascites CA-125 levels in ovarian cancer and nonovarian benign and malignant diseases: a comparative study. AB - OBJECTIVES: Cancer antigen-125 (CA-125) is not a specific tumor marker and it is synthesized by normal and malignant cells of different origins. Recently it has been shown that various diseases are associated with increased CA-125 levels, especially in the presence of serosal fluid. The aim of this study is to investigate serum and fluid CA-125 levels in patients with different diseases. METHODS: A total of 133 patients and 23 healthy control cases were included in the study and divided into eight groups on the basis of disease and the presence of fluid in the serosal cavities. Serum and serosal fluid CA-125 levels were measured by a commercial enzyme immunoassay kit at the same time. Comparisons among the groups were made. RESULTS: Abnormal levels of serum CA-125 were observed in 76% of ovarian cancer patients; 96% in patients with ascites and 56% in patients without ascites. Moreover, elevated serum CA-125 levels were detected in 52% of patients with hepatic diseases, in 100% of patients with nongynecologic peritoneal carcinomatosis, and in 87% of patients with pleural effusion. Serum and fluid CA-125 levels were significantly higher in cases of ovarian cancer with ascites than in the other groups (P < 0.01). A positive correlation between serum CA-125 levels and ascites amounts was observed in cases of ovarian cancer with ascites (P < 0.01, r = 0.81). Furthermore, no correlation was observed between ovarian mass volume and serum CA-125 levels in ovarian cancer patients with stage I disease without ascites (P = 0.08, r = 0.48). CONCLUSIONS: Although CA-125 levels may be considered a sensitive tumor marker in patients with epithelial ovarian cancer, it was determined that high serum CA-125 levels were closely related to the presence of serosal fluids and serosal involvement, whatever the origin is. These results should be considered in the interpretation of CA-125 elevation in patients with ovarian cancer. PMID- 11925129 TI - Vulvar intraepithelial neoplasia treated with cavitational ultrasonic surgical aspiration. AB - OBJECTIVE: The aim of the study was to investigate the use of the cavitron ultrasonic surgical aspirator (CUSA) for the treatment of vulvar intraepithelial neoplasia (VIN) as it combines the advantage of laser removal of the superficial dermal layers without scars and the advantage of resection with collection of a pathological specimen. METHODS: Between 1992 and 1998, 37 patients with VIN were treated using the CUSA. Charts were reviewed retrospectively. RESULTS: The median age at diagnosis was 40 years. Eleven patients (30%) had been previously treated for VIN. Diagnosis was made by inspection before and after ascitic acid application, colposcopy, and multiple biopsies revealing VIN II in 8 patients (22%) and VIN III in 29 patients (78%). At least two quadrants of the vulva were involved in 16 cases (43%) and three or four quadrants in 12 cases (33%). Under anesthesia the CUSA was used to remove all lesions with a 1-cm margin. There were no complications except 1 admission for pain control. Healing was complete in 4 to 6 weeks and no patient developed scarring. Final pathology confirmed the preoperative diagnostic grade in 24 cases (65%), while upgrading to a higher dysplasia occurred in 4 patients (11%). A second treatment was necessary in 3 patients with widespread disease. Patients were followed for an average of 33 months. Thirteen recurrences (35%) developed after a median interval of 16 months. Recurrences were significantly (P = 0.004) more frequent if VIN involved hair-bearing tissue, 6 of 7 (86%) cases, in contrast to patients with disease confined to the labia minora and introitus, 7 of 30 (23%) cases. CONCLUSION: CUSA is an acceptable treatment alternative for VIN confined to non-hair-bearing vulvar skin. PMID- 11925130 TI - Disease persistence in patients with cervical intraepithelial neoplasia undergoing electrosurgical conization. AB - OBJECTIVE: Cone margin status has been reported to be the most important predictor of residual disease in patients with cervical intraepithelial neoplasia (CIN) undergoing electrosurgical excisional treatment. The primary aim of this study of patients treated with electrosurgical conization was to evaluate the association of cone margin status and other clinical and pathologic factors with the probability of residual disease. METHODS: The study population comprised 699 patients with at least one follow-up visit within 12 months of conization. Residual disease was defined as a histology diagnosis of CIN within 3 years of conization. Multivariate associations were evaluated with multiple logistic regression analysis. RESULTS: Attendance to follow-up was 97% for the second visit and 34% for the third visit. Residual disease was detected in a total of 38 patients (5.4%). The detection rate was 3.3% at the first visit, 2.1% at the second visit, and 0.4% at the third visit. An increased probability of residual disease was associated with a referral Pap smear reported as high-grade squamous intraepithelial neoplasia and carcinoma (odds ratio, 2.9; reference category, low grade squamous intraepithelial neoplasia). A decreased probability was associated with a squamocolumnar junction entirely visible at the first follow-up visit (odds ratio, 0.2; reference category, squamocolumnar junction not visible). Patient age, time period, lesion size, lesion site, grade of CIN, cone width, cone depth, and margin status had no influence. CONCLUSIONS: The determinants of residual disease in this study differed from those generally reported so far. Factors associated with effectiveness of electrosurgical treatment of CIN need further research. PMID- 11925131 TI - Detection of female lower genital tract dysplasia using orally administered 5 aminolevulinic acid induced protoporphyrin IX: a preliminary study. AB - OBJECTIVE: Previous studies have suggested that 5-aminolevulinic acid (ALA) may be used topically on the cervix to allow optical detection of cervical dysplasia, based on the fluorescence of protoporphyrin IX (PpIX) synthesized in situ from ALA. However, the uniformity of distribution of topically applied PpIX and the sensitivity and specificity of detection are not optimal. The current study was undertaken to demonstrate the feasibility of administering ALA by mouth (po) with the hypothesis that systemic administration might provide a more reliable diagnostic tool. METHODS: Oral ALA was administered to 14 patients with abnormal Pap smears in a dose- and time-intensity design. Institutional review board approval was obtained. A starting dose of 10 mg/kg of po ALA was administered and colposcopy was performed in 3 patients at 1 h, 3 patients at 2 h, 6 patients at 3 h, and 2 patients at 4 h. The study was written with the intent to increase the dose in 10 mg/kg increments if fluorescence was not detected; however, fluorescence was detected at the first dose level. Liver function tests were checked pre and post ALA and follow-up telephone calls were made regarding possible side effects. Both white and blue light colposcopy examinations were performed by two blinded clinicians and biopsies of all abnormal areas were performed. RESULTS: All patients tolerated po ALA well, with no systemic side effects. At the 10 mg/kg dose there was no reported nausea or photosensitivity. Optimal fluorescence was achieved at the 3-h time point, with quenching noted at the 4-h time point. Excellent absorption was documented with fluorescence of the lip demonstrated with Wood's lamp. In some cases fluorescence correlated with dysplasia on biopsy. CONCLUSION: 5-ALA given via the po route and at the dose and time period studied is well tolerated and affords fluorescence of the cervix. Future study is needed to demonstrate the successful identification of dysplastic lesions, with the ultimate goal of treating dysplasia of the lower genital tract with 5-ALA and light therapy. PMID- 11925132 TI - Phase I study of alternating doublets of topotecan/carboplatin and paclitaxel/carboplatin in patients with newly diagnosed, advanced ovarian cancer. AB - OBJECTIVE: The aim of this study was to evaluate topotecan with carboplatin in an alternating doublet with carboplatin and paclitaxel in first-line ovarian cancer. METHODS: Patients with newly diagnosed stage III/IV ovarian cancer were studied. The maximum tolerated dose (MTD) of topotecan (cycles 1, 3, 5, 7) in an alternating doublet regimen was determined through standard dose escalation in cohorts of three; doses of carboplatin (area under the curve [AUC] 4 to 5) and paclitaxel (175 mg/m(2), cycles 2, 4, 6, 8) were fixed. Dose-limiting toxicity (DLT) was defined only for cycle 1 as febrile neutropenia, prolonged grade 4 granulocytopenia, grade 4 thrombocytopenia, > or =grade 3 nonhematologic toxicity, or failure to recover in < or =7 days. The use of granulocyte colony stimulating factor (G-CSF) to permit further dose escalation was also studied. RESULTS: Thirty-seven patients received 142 cycles of topotecan/carboplatin. Hematologic DLTs included grade 4 neutropenia (59 events, 42% of cycles) and thrombocytopenia (32 events, 23% of cycles). Granulocytopenia was generally short lived, and only 2 cases of febrile neutropenia occurred. The MTD was 1.0 mg/m(2)/day topotecan and carboplatin AUC 4, alternating with 175 mg/m(2) paclitaxel and carboplatin AUC 4. Although G-CSF effectively managed myelosuppression, thrombocytopenia developed in later cycles, limiting further topotecan dose escalation. The median progression-free survival was 20.5 months, and elevated pretreatment CA-125 levels normalized in 29 of 34 (85%) patients. CONCLUSION: The establishment of a reasonably well-tolerated alternating doublet regimen, coupled with evidence of antitumor activity, provides the basis for further investigation of topotecan in first-line therapy of ovarian cancer. Topotecan (1.0 mg/m(2) daily for 3 days) was chosen for further evaluation in a phase II study. PMID- 11925133 TI - A phase I trial of prolonged oral etoposide and liposomal doxorubicin in ovarian, peritoneal, and tubal carcinoma: a gynecologic oncology group study. AB - OBJECTIVES: In an effort to explore second-line therapy in ovarian, peritoneal, and tubal carcinoma, a phase I trial combining prolonged oral etoposide and liposomal doxorubicin was conducted by the Gynecologic Oncology Group. METHODS: Liposomal doxorubicin (20 mg/m(2)) was administered intravenously over 1 h followed by oral etoposide at 50 mg/m(2)/day beginning on day 2. In the first phase of accrual, the number of days of oral etoposide was increased until its maximum tolerated dose (MTD) was determined based on hematologic toxicity. In the second phase, etoposide was given at the MTD while the dose of liposomal doxorubicin was escalated until its maximum tolerated dose was reached based on hematologic or nonhematologic toxicity. Cycles were repeated every 28 days for a maximum of 12 courses. Dose-limiting toxicity was defined as neutropenic sepsis, grade 4 thrombocytopenia, absolute neutrophil count <1000/microl or platelets <50,000 during treatment with etoposide, or > or =grade 3 mucositis/stomatitis, palmar-plantar erythrodyesthesia, or rash. RESULTS: Fifteen patients were accrued to the study's first phase, and 3 were accrued to the second phase. Dose-limiting hematologic toxicity occurred with 14 days of oral etoposide in combination with liposomal doxorubicin at 20 mg/m(2). Efforts to escalate the dose of liposomal doxorubicin to 30 mg/m(2) in combination with 12 days of oral etoposide at 50 mg/m(2) resulted in dose-limiting hematologic toxicity. Five of 17 (29%; 95% CI: 13-53%) evaluable patients experienced a response. CONCLUSION: The regimen of oral etoposide at 50 mg/m(2)/day for 12 days in combination with liposomal doxorubicin at a dose of 20 mg/m(2) is tolerable without supportive therapy. While this dose of oral etoposide has demonstrated activity as a single agent in ovarian cancer, liposomal doxorubicin has only been effective in ovarian cancer at higher doses. There are no immediate plans to study this combination further. PMID- 11925134 TI - Prospective assessment of quality of life of female cancer patients. AB - OBJECTIVE: The aims of this study were to compare the quality of life (QOL) of women with different cancer sites; to identify predictors of QOL; and to examine the agreement between patient self-reported QOL and QOL ratings provided by clinicians and significant others. METHODS: A prospective study was conducted including 248 patients with gynecologic and breast cancer. QOL data were collected at six time points before, during, and after treatment, using the EORTC QLQ-C30 and the Spitzer QL index (QL-I). RESULTS: Baseline assessments showed comparable QOL scores among patients with different gynecologic malignancies and breast cancer. During active treatment breast cancer patients had significantly higher mean scores in physical functioning compared to women with gynecologic cancers and higher scores in role functioning compared to patients with cervical cancer. After completion of treatment there were no statistically significant differences in QOL among the groups. For all women, global QOL and emotional functioning were mostly affected during and after treatment. Regression analysis showed that patients' global QOL was significantly predicted by severity of surgery (t = 3.903, P < 0.01) and pretreatment performance status (t = 3.116, P = <0.01). Comorbidity, family support, number of treatments, age, and stage of disease were not predictive. The comparison of patient self-rated QOL and observer-rated QOL showed that the QL-I mean scores of health providers and relatives were generally in close agreement with those of patients. Intraclass correlations were moderate to high during active treatment and excellent after completion of treatment. CONCLUSION: In female cancer patients, global QOL and emotional functioning are mostly affected during the course of disease, independent of their diagnosis. Significant others and health professionals are able to provide useful information on QOL of patients recovering from cancer. PMID- 11925136 TI - Modified radical vaginal hysterectomy with or without laparoscopic nerve-sparing dissection: a comparative study. AB - OBJECTIVE: Nerve-sparing dissection of the lateral portion of the cardinal ligament (paracervical lymphadenectomy) has been recently developed with the goal of reducing the late urinary adverse effects of radical hysterectomy without impairing the outcome. This work has been carried out in order to investigate the influence of additional paracervical lymph node dissection at the time of laparoscopically assisted modified radical hysterectomy on outcome and urinary sequelae. METHODS: A total of 95 patients underwent a modified radical hysterectomy using a combined laparoscopic and vaginal approach since 1991. In 47 of these patients treated since 1996 an additional laparoscopic paracervical lymphadenectomy was performed. The operative, postoperative, and survival outcomes were assessed. In 60 patients, 32 and 28 in the groups of patients with or without paracervical dissection, respectively, a careful interview on urinary symptoms was conducted by an independent investigator. RESULTS: Paracervical dissection involves no operative complication and lenghthens the postoperative urinary retention, but has no adverse influence on long-term urinary discomfort. The yield of paracervical dissection is negligible for small tumors: no positive node was found in 38 patients with tumors less than 2 cm in diameter. The outcome of patients after minimal access surgery for tumors less than 2 cm is excellent whether or not a paracervical dissection has been performed. CONCLUSION: Paracervical dissection does not worsen the late urinary symptoms after vaginal radical hysterectomy. It has a limited value in the surgical management of small size (less than 2 cm) cervical cancers, although it may prevent long-term lateropelvic recurrences. PMID- 11925135 TI - Detecting cytokeratin 19 mRNA in the peripheral blood cells of cervical cancer patients and its clinical-pathological correlation. AB - OBJECTIVE: The aim of this study was to study the presence of cytokeratin 19 (CK19)-expressing cancer cells in the blood of preoperative patients with FIGO stage Ib and IIb cervical cancers who received radical hysterectomy and to investigate the cells' clinical significance. METHODS: CK19 mRNA in the blood cells of the patients was detected preoperatively by a newly designed nested reverse transcriptase-polymerase chain reaction, which excluded pseudogenes a and b, performed on 84 patients with stage Ib and IIb cervical carcinoma. Possible correlations between clinicopathological factors were then analyzed. RESULTS: The sensitivity of this assay was 1 CK19-mRNA-positive cell per 10(6) peripheral blood mononuclear cells. Results showed that 21.4% of the 84 patients with cervical carcinoma had CK19-mRNA-positive cells in the blood, in comparison with 5.7% of the 35 patients with benign gynecological tumors and 0% of the 28 healthy controls (P = 0.037 and 0.006, respectively). The positive tests in the cervical cancer patients were not associated with prognostic factors including stage, pelvic lymph node metastasis, pathological types, bulky tumor size (> or =4 cm), differentiation, parametrial extension, lymphovascular space involvement, deep stromal invasion, or age. CONCLUSIONS: This study revealed the presence of circulating CK19-expressing cancer cells in the blood of patients with untreated early-stage cervical carcinomas, indicating that cervical cancer disseminated early. The survival effect of this phenomenon must be clarified. This detection assay provides an early checkpoint in the multistep process for developing metastasis in cervical cancer patients. PMID- 11925137 TI - Muscarinic receptors in cell lines from ovarian carcinoma: negative correlation with survival of patients. AB - OBJECTIVES: Tumor cells are similar in many respects to embryonic cells, indicating that embryonic genes are reactivated during malignant growth. In previous studies, we observed muscarinic acetylcholine receptors, which are expressed in embryonic cells during morphogenesis and are also found in human melanomas and melanoma cell lines. We determined the presence of muscarinic receptors in a collection of ovarian tumor cell lines for which clinical data were available. METHODS: Muscarinic receptor status of 39 cell lines derived from 34 patients was determined by Western blotting. RESULTS: Twenty-three cell lines were receptor positive, and 16, receptor negative. Kaplan-Meier analysis of receptor status of the tumor cell lines and survival time of patients from which the cell lines were established showed that expression of muscarinic receptors was associated with a reduced probability (P = 0.025) of survival: This is within the range of other established prognostic factors reported in the literature. CONCLUSIONS: A large percentage of ovarian tumor cell lines express muscarinic receptors. Muscarinic receptor expression is an embryonic trait and is correlated with reduced survival of patients. The results from this study provide further evidence of the involvement of muscarinic receptors in the progression of malignant carcinomas. PMID- 11925138 TI - PTEN mutations in uterine sarcomas. AB - OBJECTIVE: Uterinesarcomas comprise three main types: carcinosarcomas, leiomyosarcomas, and endometrial stromal sarcomas. Carcinosarcomas are highly aggressive neoplasms with a biphasic histology of carcinomatous and sarcomatous elements. It is now generally accepted that carcinosarcomas are biphasic tumors that have to be regarded as endometrial carcinomas where metaplasia occurs. Mutations of the PTEN tumor suppressor gene, located on 10q23, play a significant role in the pathogenesis of the endometrioid type of endometrial carcinoma. Loss of heterozygosity of chromosome 10q has been reported in uterine leiomyosarcoma. Since little is known about the molecular pathobiology, our goal was to investigate the potential role of the PTEN gene in the carcinogenesis of uterine sarcomas. METHODS: We examined 21 carcinosarcomas, 21 leiomyosarcomas, and 5 endometrial stromal sarcomas using exon-by-exon polymerase chain reaction-single strand conformation polymorphism analysis. RESULTS: Overall 8.5% (4/47) of uterine sarcomas were found to harbor somatic PTEN mutations. Of these, approximately 17% (3/18) were carcinosarcomas with endometrioid-type carcinoma components and approximately 5% (1/21) were leiomyosarcomas. No mutations were detected in carcinosarcomas with nonendometrioid carcinoma components (0/3) and in endometrial stromal sarcomas (0/5). CONCLUSIONS: These data suggest that intragenic PTEN mutations are involved in the genesis of uterine carcinosarcomas with endometrioid-type carcinoma components but rarely contribute to the pathobiology of uterine leiomyosarcomas. PMID- 11925139 TI - Infrared spectral features of exfoliated cervical cells, cervical adenocarcinoma tissue, and an adenocarcinoma cell line (SiSo). AB - OBJECTIVE: The objective of the present study is to evaluate and compare the infrared spectral features of normal and malignant exfoliated cervical cells, cells from malignant tissue, and the SiSo cell line. METHODS: Infrared spectra of cervical adenocarcinoma (CA) tissue, normal and malignant exfoliated cervical cells, and a uterine cervical adenocarcinoma cell line (SiSo) were obtained. Spectral qualities in terms of band intensity ratio and band position, which reflect configurational changes in the functional groups of the above samples, were measured. RESULTS: Spectral bands of CA tissue, exfoliated cells from CA, and the cell line were similar but markedly different from that of exfoliated normal cervical cells. Significant changes in bands at 1025 cm(-1) (glycogen), 1080 cm(-1) (glycogen and nucleic acids), 1155 cm(-1) (C-OH groups of serine, threonine, and tyrosine of cell proteins, and C-O groups of carbohydrates), 1240 cm(-1) (PO(2) groups of nucleic acids), 1400 cm(-1) (methyl group of lipids and proteins), and 1450 cm(-1) (methylene group of lipids and proteins) were noted in the CA tissue, exfoliated CA cells, and adenocarcinoma cell line compared with exfoliated normal cells. Marked shifts in band positions from 1080 to 1086 cm( 1), 1153 to 1160 cm(-1), and 935 to 970 cm(-1) in CA tissue, exfoliated CA cells, and the adenocarcinoma cell line were noted. CONCLUSION: Spectral bands of the adenocarcinoma cell line matched very well with those of cervical CA tissue and exfoliated CA cells in terms of position. In contrast, spectral bands of the SiSo cell line differed greatly from those of normal exfoliated cells. PMID- 11925140 TI - The significance of serum CA 125 elevation in malignant and nonmalignant diseases. AB - OBJECTIVES: The aims of the study were to investigate whether an elevated CA 125 level signals malignancies other than ovarian cancer and to find the cause of death for 247 women with elevated values among the 5550 women screened in 1986 1988 in the Stockholm population. METHODS: The Swedish Regional Cancer Registry delivered malignancy diagnoses among the 5550 women screened. The Cause of Death Registry gave the cause of death among the women with elevated CA 125 values. RESULTS: Patients with ovarian cancer were excluded. In 44 women with elevated CA 125 values other malignancies were reported to the Cancer Registry. They represent 18% of the entire group with elevated values. Among the 5297 women with normal CA 125 values 13% developed various malignancies. The difference between incidence of malignant disease in women with elevated values and women with normal values is significant, P = 0.02. Especially during the test-related period, from 1 year before to 1 year after the test, malignancies were detected in 6.9% of the population with elevated values and in only 1.6% with normal values (P = < 0.001). Breast cancer and lung cancer were overrepresented among women with elevated CA 125 values (P = 0.015 and < 0.001, respectively). Of the total 5500 women screened, 358 women died with different diagnoses. Elevated CA 125 values had been noted earlier in 25 women, and of these 20 died of malignant diseases, predominantly ovarian, breast, and lung cancer. CONCLUSIONS: Asymptomatic postmenopausal women with elevated CA 125 levels in ovarian carcinoma screening trials should, if ovarian cancer is excluded, be investigated for possible breast or lung cancer. The findings also suggest that elevated CA 125 level is a risk factor for death from malignant disease. PMID- 11925141 TI - Prospective evaluation of positron emission tomography for the detection of groin node metastases from vulvar cancer. AB - OBJECTIVE: We set out to determine the ability of positron emission tomography with fluorodeoxyglucose to detect groin lymph node metastases from vulvar cancer. METHODS: From January 2000 to August 2001, patients with squamous cell cancer of the vulva undergoing radical excision and lymphadenectomy were offered preoperative positron emission tomography. The imaging and pathologic status of each patient and groin were compared, and the sensitivity, specificity, and predictive value of positron emission tomography in predicting nodal metastasis were determined. RESULTS: Fifteen patients underwent positron emission tomography prior to exploration of 29 groins. Six patients had positive scans, suggesting metastases in 8 groins. Pathologically, 5 patients had metastases in 9 groins, with positron emission tomography demonstrating metastases in 4 of 5 patients and 6 of 9 groins with disease. On a patient-by-patients basis, positron emission tomography had a sensitivity of 80%, specificity of 90%, positive predictive value of 80%, and negative predictive value of 90% in demonstrating metastases. On a groin-by-groin basis, positron emission tomography had a sensitivity of 67%, specificity of 95%, positive predictive value of 86%, and negative predictive value of 86%. Positron emission tomography was more accurate in detecting extranodal metastases than disease confined within the groin nodes (P = 0.048). CONCLUSIONS: Positron emission tomography is relatively insensitive in predicting lymph node metastasis, and a negative study is not a reliable surrogate for a pathologically negative groin. However, the high specificity suggests that positron emission tomography is useful in planning radiation therapy and as an adjunct to lymphatic mapping and sentinel lymph node dissection. PMID- 11925142 TI - Phase I study of topotecan and radiation therapy in advanced cervical cancer. AB - OBJECTIVE: The goal of this study was to determine the maximal tolerated dose (MTD) of topotecan given with external-beam radiotherapy in advanced cervical cancer. METHODS: A prospective Phase I trial of topotecan given with standard external-beam radiotherapy was performed in patients with advanced squamous cell carcinoma of the cervix. Patients were treated with a starting dose of 0.5 mg/m(2) and escalated by 0.25 mg/m(2). Nine patients were treated. Hematologic and nonhematologic toxicity were measured. RESULTS: Patients were treated with 1.0 mg/m(2) daily for 5 days on Days 1-5 and 22-26 concomitantly with radiotherapy without significant toxicity. Grade III anemia in one case and Grade II leukopenia in two cases were seen in the three patients at this dose level. Dose-limiting toxicity was not reached. CONCLUSION: Topotecan can be safely administered at a dose of 1.0 mg/m(2) during external-beam radiotherapy for advanced cervical cancer. PMID- 11925143 TI - Rectus femoris myocutaneous flap for vulvoperineal reconstruction. AB - BACKGROUND: Many reconstructive procedures for large vulvoperineal defects have been described. The present report describes the use of the rectus femoris myocutaneous flap, which has not previously been described for this purpose. CASE: A 52-year-old woman suffered a local recurrence of a Bartholin's gland carcinoma after anterior exenteration and pubectomy. A palliative resection was performed which resulted in a large vulvoperineal defect with transpelvic herniation of the peritoneal contents. This was immediately reconstructed with a rectus femoris myocutaneous flap. Her postoperative course and healing were uneventful. CONCLUSION: This technique is an alternative method for vulvar reconstruction. It is especially useful for large defects when a gracilis or rectus abdominis flap is not available. PMID- 11925144 TI - Use of medroxyprogesterone acetate in the treatment of Mullerian adenosarcoma: a case report. AB - BACKGROUND: Mullerianadenosarcoma is a rare pelvic malignancy that most commonly arises from the endometrium. These tumors are relatively insensitive to chemotherapy and radiation and are primarily treated by surgical resection. We report a case of mullerian adenosarcoma arising outside of the uterus from a background of endometriosis treated with a combination of surgical resection and medroxyprogesterone acetate. CASE: A 43-year-old woman with a history of endometriosis was diagnosed with advanced extrauterine mullerian adenosarcoma. After suboptimal tumor dubulking surgery she was treated with medroxyprogesterone acetate. Ten months postoperatively she remains without evidence of disease. CONCLUSION: Medroxyprogesterone acetate may be a useful drug in the treatment of advanced mullerian adenosarcoma. PMID- 11925145 TI - Positron emission tomography for detecting clinically occult surgically resectable metastatic ovarian cancer. AB - BACKGROUND: The specific indications for PET imaging in patients with ovarian cancer have yet to be precisely defined. While PET has limited sensitivity for detecting small-volume (<1 cm) metastatic disease, accurate identification of larger tumor nodules may have a significant impact on clinical management and the selection of patients for cytoreductive surgery. CASES: The cases of two patients with suspected recurrent Stage IIIC serous ovarian cancer based solely on elevated CA125 levels and one patient with an apparent Stage IC poorly differentiated ovarian sex cord-stromal tumor who had macroscopic surgically resectable disease (>1 cm) identified by PET after negative or equivocal computed tomography are presented. CONCLUSION: PET imaging may be a useful technique for identifying potentially surgically resectable, macroscopic metastatic ovarian cancer when computed tomography findings are negative or equivocal. PMID- 11925146 TI - Successful laparoscopic removal of a solitary adrenal metastasis from ovarian carcinoma: a case report. AB - BACKGROUND: The adrenal gland is a site of metastatic spread of some tumors. Usually the presence of adrenal metastasis signals widespread disease; however, isolated adrenal metastases have been reported. The adrenal gland, along with certain other organs, is considered a pharmacological sanctuary. Several reports describe long-term survival after adrenalectomy due to isolated adrenal metastases, in particular from non-small-cell lung cancer. There are several reports of laparoscopic resection of isolated adrenal metastases. CASE: We present a rare case of a laparoscopically resected solitary adrenal metastasis originating from ovarian carcinoma. To the best of our knowledge there was no report in the literature of an isolated adrenal metastasis from ovarian carcinoma. The patient described developed an isolated left adrenal metastasis 11 months after completion of adjuvant chemotherapy for stage III ovarian cancer. The serum CA125 dropped from 365 to 35 postoperation. CONCLUSION: Isolated adrenal metastasis from ovarian cancer is a rare event, never before described in the literature. This case report describes a laparoscopic resection of an adrenal metastasis from ovarian cancer, and the drop of CA125 levels indicated complete resection of the metastasis. PMID- 11925147 TI - Choriocarcinoma and epithelial trophoblastic tumor: successful treatment of relapse with hysterectomy and high-dose chemotherapy with peripheral stem cell support: a case report. AB - BACKGROUND: Post-termchoriocarcinoma is a rare complication of pregnancy. The presence of epithelioid trophoblastic elements may lead to the persistence of locally invasive disease which is unresponsive to multiagent chemotherapy. Relapse is associated with an estimated mortality rate of 30%. CASE: We present a case of Stage IC post-term choriocarcinoma and epithelioid trophoblastic tumor. While the metastatic sites in the lungs responded to multiagent chemotherapy, a hysterectomy was required to treat persistent disease in the uterus. The patient relapsed within 4 months of completion of chemotherapy. Relapse was treated with high-dose chemotherapy with peripheral stem cell support. The patient is alive with no evidence of disease 23 months posttransplant. CONCLUSIONS: The application of multimodality treatment and high-dose chemotherapy resulted in a successful outcome for this patient, indicating a potential role for high-dose therapy in patients who suffer a relapse of choriocarcinoma. PMID- 11925148 TI - Cutaneous metastasis from carcinoma of the cervix at the drain site. AB - BACKGROUND: Metastasis to the skin occurs rarely in gynecologic cancer, especially in cervical carcinoma. Although carcinoma of the cervix is the second to the fourth most common malignancy in women, cutaneous involvement originating from cervical cancer is particularly unusual, even in the terminal stage of the disease. CASE: We present a case of cervical cancer recurrence with skin metastasis. The extensive skin lesion on the abdominal wall occurred 4 years after radical surgery and postoperative radiotherapy. This was a biopsy-proven metastasis from the patient's primary cervical carcinoma. CONCLUSION: As far we know this is the second case (after Copas et al., Gynecol Oncol 1995;56:102-4) of skin and subcutaneous tissues metastasis from cervical carcinoma at the drain site. Palliative chemotherapy and radiotherapy have a useful role in controlling symptoms. PMID- 11925149 TI - Endovascular management of ureteral-iliac artery fistulae with Wallgraft endoprostheses. AB - BACKGROUND: Ureteral-iliac artery fistulae are rare, yet potentially life threatening, causes of hematuria. Treatment has traditionally been surgical, but advances in endovascular technology have led to a few recent reports of therapy with covered stents. We report two cases of patients diagnosed with ureteral iliac artery fistulae who were treated with Wallgraft endoprostheses, a new, commercially available covered stent. CASES: We report two patients with gynecologic malignancies who presented with massive hematuria and hypotension and were subsequently proven to have ureteral-iliac arterial fistulae. Both patients had prior pelvic surgery, radiation, and chronic indwelling ureteral stents. Once the diagnosis was established, both patients were managed with endovascular covered stent placement. The patients' conditions stabilized, hematuria ceased, and both were discharged from the hospital without additional transfusion or surgical treatment. CONCLUSION: Endovascular therapy with covered stents is a safe, effective, and readily available method for the treatment of ureteral-iliac artery fistulae. PMID- 11925150 TI - Vulvar epithelioid sarcoma in pregnancy. AB - BACKGROUND: Epithelioid sarcoma is a soft tissue tumor rarely found centrally and even less commonly on the vulva. Vulvar sarcoma in pregnancy is also exceedingly rare with only five cases reported to date, none of which have been an epithelioid sarcoma. CASE: We report a case of a 29-year-old woman presenting with a vulvar epithelioid sarcoma at 36 weeks of gestation. The patient underwent a radical resection 6 weeks postpartum followed by chemotherapy. Despite a radical hemivulvectomy and doxorubicin and ifosfamide chemotherapy, she developed pulmonary metastasis and died of tumor-related pulmonary failure secondary to her disease 612 months after diagnosis. To our knowledge this is the first case of a vulvar epithelioid sarcoma presenting during pregnancy. The English literature is reviewed and a total of 18 previous cases of vulvar epithelioid sarcoma have been reported outside of pregnancy. Insight into the biological behavior and therapeutic management of this disease is discussed. CONCLUSION: The optimal management of vulvar epithelioid sarcoma remains to be determined. However, it would seem that early and aggressive surgical resection provides the best possibility for cure. The role of radiation and/or chemotherapy remains to be determined. PMID- 11925152 TI - The role of less radical surgery in selected cervical cancers. PMID- 11925151 TI - Retinoids enhance cisplatin-based chemoradiation in cervical cancer cells in vitro. AB - OBJECTIVE: The aim of this study was to determine whether receptor-dependent and receptor-independent retinoids sensitize cervical cancer cells to clinically relevant doses of concurrent radiation and cisplatin. METHODS: The clonogenic assay was performed on SiHa cervical carcinoma cultures treated with 5 microM 9 cis-retinoic acid (RA) or 3 microM 4-HPR for 3 days prior to and following concurrent treatment with 3 microM cisplatin and 2 Gy of Co(60) radiation. RESULTS: Neither 9-cis-RA nor 4-HPR significantly decreased survival for radiation only or cisplatin only (t test: P < 0.05), but both significantly decreased survival of cultures receiving concurrent chemoradiation (t test: 9-cis RA P = 0.045; 4-HPR P = 0.027). CONCLUSIONS: Both receptor-dependent and receptor independent retinoids enhance concurrent chemoradiation effects in vitro. PMID- 11925154 TI - Choices in creating continent urostomies following pelvic exenteration for gynecologic malignancies. PMID- 11925155 TI - A new high-yield synthesis of Cl(3)P=NSiMe(3), a monomeric precursor for the controlled preparation of high molecular weight polyphosphazenes. AB - The phosphoranimine, Cl(3)P=NSiMe(3), was prepared using a new, high-yield (>80%), one-pot synthesis via oxidation of the chlorophosphine, Cl(2)PN(SiMe(3))(2) with SO(2)Cl(2) in ether. Cl(3)P=NSiMe(3) is a valuable monomeric precursor in the synthesis of well-defined polyphosphazenes. PMID- 11925156 TI - Precipitation of a dioxouranium(VI) species from a room temperature ionic liquid medium. AB - The novel complex 1-butyl-3-methylimidazolium mu(4)-(O,O,O',O'-ethane-1,2-dioato) bis[bis(nitrato-O,O)dioxouranate(VI)] (1) has been precipitated from a room temperature ionic liquid medium containing 1-butyl-3-methylimidazolium nitrate, nitric acid, and acetone. X-ray analysis of complex 1 shows the unit cell contains four [C(4)mim](+) cations and two independent [[UO(2))(NO(3))(2)](2)(mu(4)-C(2)O(4))](2-) moieties, both of which are located about inversion centers. The [C(4)mim](+) cations are arranged such that they produce large channels in which the anions are located. This arrangement of [[(UO(2))(NO(3))(2)](2)(mu(4)-C(2)O(4))](2-) groups is unique to this compound. Crystal data for compound 1: M = 1154.56, monoclinic, space group P2(1)/c, a = 15.452(2) A, b = 20.354(3) A, c = 10.822(4) A, beta = 106.84(2) degrees, U = 3258(1) A(-)(3), Z = 4, mu = 10.023 mm(-1), R(int) = 0.0788. PMID- 11925157 TI - Selective cleavage of P-N bonds and the conversion of rhodium N-pyrrolyl phosphine complexes into diphosphoxane-bridged dimers. AB - Rhodium(I) complexes trans-[RhCl(CO)(PR(2)[NC(4)H(3)C(O)Me-2])(2)] (R = Ph, NC(4)H(4)) react with water to give the diphosphoxane-bridged dimers [Rh(2)Cl(2)(CO)(2)(mu-PR(2)OPR(2))(2)] following cleavage of the P-N bonds to the 2-acetyl-N-pyrrolyl groups. The two dimers have been crystallographically characterized and show a number of structural differences, with the PPh(2)OPPh(2) compound possessing semibridging chloride and carbonyl ligands whereas the P(NC(4)H(4))(2)OP(NC(4)H(4))(2) compound contains only terminal chlorides and carbonyls. No evidence for cleavage of the P-N bonds involving the unfunctionalized N-pyrrolyl groups in trans [RhCl(CO)(P[NC(4)H(4)](2)[NC(4)H(3)C(O)Me-2])(2)] was observed. PMID- 11925158 TI - Observation of the structural consequences of a reversible S = 3/2 and S = 1/2 spin crossover in the single crystal. AB - Highly saddle shaped iron(III) porphyrin complex 1 showing a novel spin crossover process between the S = 3/2 and S = 1/2 has been crystallographically analyzed at 298, 180, and 80 K. As the temperature is lowered, the lattice contraction has occurred specifically along the b-axis. Correspondingly, the iron-pyridine bonds, which tilt slightly from the b-axis, have decreased by 7.3%. In contrast, the lattice contractions along the a- and c-axes are much smaller and the iron porphyrin bonds, which almost coincide with the a- and c-axes, have shown much smaller contraction, ca. 2.2%. The large contraction of the specific bonds caused by packing force raises the energy level of the d(z)2 orbital and induces the spin transition. The detailed structural and lattice changes in the single crystal, which may be regarded as a superstructure parallel to a protein matrix, have been clarified. PMID- 11925159 TI - CrCpTp: a high-spin Cr(II) sandwich complex with a large structural distortion. AB - CrCpTp was synthesized from the reaction of [CrCpCl](2) and KTp. Magnetic measurements indicate it to have a high-spin (S = 2) electronic configuration from 5 to 300 K. A single-crystal X-ray study reveals bond lengths typical for a high-spin configuration and a pronounced Jahn-Teller distortion. The nature of this distortion was probed by DFT calculations and the variation in bond lengths successfully reproduced. The metal-based HOMO is significantly antibonding with respect to a single pyrazolyl ring only. PMID- 11925160 TI - Synthesis of Ln(III) chloride tetraphenylporphyrin complexes. AB - The isolation and identification of the first examples of anhydrous lanthanide chloride tetraphenylporphyrin complexes have been described. The purple complexes were generated by the reaction of dilithiotetraphenylporphyrin bis(dimethoxyethane) with lanthanide trichloride tris(tetrahydrofuran) salts to yield the products in up to 85% yield. The crystal structures for the holmium and ytterbium complexes are also presented. PMID- 11925161 TI - Enhanced selectivity for actinides over lanthanides with CMPO ligands secured to a C(3)-symmetric triphenoxymethane platform. AB - A ligand system containing three preorganized carbamoylmethylphosphine oxide (CMPO) moieties anchored onto a rigid C(3)-symmetric triphenoxymethane platform has been developed for facile metal complexation and subsequent extraction from aqueous acidic nuclear waste streams. Intended to mimic the 3:1 CMPO-actinide stoichiometry of the extracted species in the TRUEX nuclear waste treatment process, the CMPO arms on this ligand are oriented such that all three CMPO moieties can cooperatively bind a metal ion. Extractions of simulated nuclear waste streams (10(-4) M metal in 1 M nitric acid) with solutions of this ligand in methylene chloride (10(-3) M) reveal a high affinity for the actinide thorium and a very low, but constant, affinity for the lanthanides across the series. Thorium and five lanthanide (lanthanum, cerium, neodymium, europium, and ytterbium) nitrate complexes of this ligand have been synthesized and fully characterized by X-ray crystallography, (1)H and (31)P NMR spectra, and FT-ICR-MS to elucidate the mechanism of this unique actinide selectivity. All six oxygen donors from the three CMPO arms of the ligand and one or two nitrate counterions coordinate these metals to afford 2+ cationic complexes in every case. Because of the large size of the ligand, both the thorium and lanthanide complexes present similarly charged and sized surfaces to the extraction solvents, but the thorium complex is extracted quantitatively over the lanthanide complexes. A possible rationale for this extraction behavior difference is presented and further illustrated by the extraction properties of this ligand system for the alkali metals (lithium, sodium, potassium, rubidium, and cesium) as picrate salts and by the solid- and solution-state structures of its lithium picrate complex. PMID- 11925162 TI - Mechanistic diversity covering 15 orders of magnitude in rates: cyanide exchange on [M(CN)(4)](2-) (M = Ni, Pd, and Pt). AB - Kinetic studies of cyanide exchange on [M(CN)(4)](2-) square-planar complexes (M = Pt, Pd, and Ni) were performed as a function of pH by (13)C NMR. The [Pt(CN)(4)](2-) complex has a purely second-order rate law, with CN(-) as acting as the nucleophile, with the following kinetic parameters: (k(2)(Pt,CN))(298) = 11 +/- 1 s(-1) mol(-1) kg, DeltaH(2) (Pt,CN) = 25.1 +/- 1 kJ mol(-1), DeltaS(2) (Pt,CN) = -142 +/- 4 J mol(-1) K(-1), and DeltaV(2) (Pt,CN) = -27 +/- 2 cm(3) mol(-1). The Pd(II) metal center has the same behavior down to pH 6. The kinetic parameters are as follows: (k(2)(Pd,CN))(298) = 82 +/- 2 s(-1) mol(-1) kg, DeltaH(2) (Pd,CN) = 23.5 +/- 1 kJ mol(-1), DeltaS(2) (Pd,CN) = -129 +/- 5 J mol( 1) K(-1), and DeltaV(2) (Pd,CN) = -22 +/- 2 cm(3) mol(-1). At low pH, the tetracyanopalladate is protonated (pK(a)(Pd(4,H)) = 3.0 +/- 0.3) to form [Pd(CN)(3)HCN](-). The rate law of the cyanide exchange on the protonated complex is also purely second order, with (k(2)(PdH,CN))(298) = (4.5 +/- 1.3) x 10(3) s( 1) mol(-1) kg. [Ni(CN)(4)](2-) is involved in various equilibrium reactions, such as the formation of [Ni(CN)(5)](3-), [Ni(CN)(3)HCN](-), and [Ni(CN)(2)(HCN)(2)] complexes. Our (13)C NMR measurements have allowed us to determine that the rate constant leading to the formation of [Ni(CN)(5)](3-) is k(2)(Ni(4),CN) = (2.3 +/- 0.1) x 10(6) s(-1) mol(-1) kg when the following activation parameters are used: DeltaH(2)() (Ni,CN) = 21.6 +/- 1 kJ mol(-1), DeltaS(2) (Ni,CN) = -51 +/- 7 J mol( 1) K(-1), and DeltaV(2) (Ni,CN) = -19 +/- 2 cm(3) mol(-1). The rate constant of the back reaction is k(-2)(Ni(4),CN) = 14 x 10(6) s(-1). The rate law pertaining to [Ni(CN)(2)(HCN)(2)] was found to be second order at pH 3.8, and the value of the rate constant is (k(2)(Ni(4,2H),CN))(298) = (63 +/- 15) x10(6) s(-1) mol(-1) kg when DeltaH(2) (Ni(4,2H),CN) = 47.3 +/- 1 kJ mol(-1), DeltaS(2) (Ni(4,2H),CN) = 63 +/- 3 J mol(-1) K(-1), and DeltaV(2) (Ni(4,2H),CN) = - 6 +/- 1 cm(3) mol( 1). The cyanide-exchange rate constant on [M(CN)(4)](2-) for Pt, Pd, and Ni increases in a 1:7:200 000 ratio. This trend is modified at low pH, and the palladium becomes 400 times more reactive than the platinum because of the formation of [Pd(CN)(3)HCN](-). For all cyanide exchanges on tetracyano complexes (A mechanism) and on their protonated forms (I/I(a) mechanisms), we have always observed a pure second-order rate law: first order for the complex and first order for CN(-). The nucleophilic attack by HCN or solvation by H(2)O is at least nine or six orders of magnitude slower, respectively than is nucleophilic attack by CN(-) for Pt(II), Pd(II), and Ni(II), respectively. PMID- 11925163 TI - Two-electron transfer for Tl(aq)(3+)/Tl(aq)(+) revisited. Common virtual [Tl(II) Tl(II)](4+) intermediate for homogeneous (superexchange) and electrode (sequential) mechanisms. AB - Homogeneous and electrochemical two-electron transfers within the Tl(aq)(3+)/Tl(aq)(+) couple are considered on a common conceptual basis. For the 2 equiv electrochemical reduction of Tl(aq)(3+) to Tl(aq)(+), the intermediate state with a formal reduction potential, E(1) = 1.04 +/- 0.10 V vs the normal hydrogen electrode, was detected, different from the established value of 0.33 V for a Tl(3+)/Tl(2+) couple. Examination of obtained electrochemical (cyclic voltammetry (CV) and rotating disk electrode techniques, along with the CV-curve computer simulation procedure) and literature data indicate that the detected formal potential cannot be the property of electrode-adsorbed species, but rather of the covalently interacting dithallium intermediate [Tl(II)-Tl(II)](4+) located at the outer Helmholtz plane. The analysis of microscopic mechanisms, based on the recent hypothesis of H. Taube and the Marcus-Hush theory extended by Zusman and Beratan, and Koper and Schmickler, revealed that the homogeneous process most probably takes place through the superexchange inner-sphere two-electron-transfer mechanism, via an essentially virtual (undetectable) dithallium intermediate. In contrast, the electrochemical process occurs through a sequential mechanism, via the rate-determining step of Tl(aq)(2+) ion formation immediately followed by activationless formation of the metastable (CV-active) dithallium state. The second electrochemical electron-transfer step is fast, and shows up only in the peak height (but not in the shape) of the observed CV cathodic wave. The anodic wave for a microscopically reverse process of the oxidation of Tl(aq)(+) to Tl(aq)(3+) cannot be observed within the considered potential range due to the blocking of through-space electron transfer by the competitor process of ion transfer to the electrode. PMID- 11925164 TI - Bonding behavior of Co(CO)(3)L (L = CO, PPh(3)) building blocks in platinum cobalt carbonyl clusters. AB - The reaction of [Co(CO)(4)](-) with [Pt(2)Cl(mu-PPh(2))(PPh(3))(3)] (1:1 ratio), which was prepared in situ by reaction of aqueous HCl with the orthometalated complex [Pt(2)(mu-PPh(2))(mu-o-C(6)H(4)PPh(2))(PPh(3))(2)], afforded the deep green triangular cluster [Pt(2)Co(mu-PPh(2))(CO)(4)(PPh(3))(2)], 7. X-ray crystallographic analysis reveals that this new cluster contains two formally monoanionic fragments, PPh(2-) and Co(CO)(4-), that bridge a d(9)-d(9) Pt(I) Pt(I) metal-metal bond. Whereas tetracarbonylcobaltate is generally bonded to only one metal center as a 2e donor ligand, it is best viewed here as a formally 4e donor anionic metalloligand. This model leads then to the usual 16e count for each Pt center and relates this metalloligand to other anionic bridging ligands, such as PPh(2-). A comparative EHMO bonding analysis of [Co(CO)(3)L](-) (L = CO, PR(3)) fragments is presented which takes into account the possible coordination geometries about cobalt, trigonal bipyramidal (with C(3v) local symmetry) or edge capped tetrahedral when referring to the ligand polyhedron (with C(2v) or C(s) symmetry). The results support the description of this unusual bridging bonding mode. PMID- 11925165 TI - Substitution reactions on cyclometalated Pt(IV) complexes. Associative tuning by fluoro ligands and fluorinated substituents. AB - The substitution reactions of sulfide by phosphines on Pt(IV) complexes having a cyclometalated imine ligand, two methyl groups in a cis geometrical arrangement, and a halogen and a sulfide as ligands, [Pt (Me)(2)X(C-N)(SR(2))], have been studied as a function of temperature, solvent, and electronic and steric characteristics of the phosphines, sulfides, X, and C-N. In most of these cases, a limiting dissociative mechanism has been found, where the dissociation of the sulfide ligand corresponds to the rate-determining step. The intermediate species formed behaves as a true pentacoordinated Pt(IV) compound in a steady-state concentration only for the systems with SMe(2); for the bulkier SEt(2) and SBzl(2) leaving ligands the rate constants and activation parameters show an important degree of solvent dependence, which correlates with the ability of the solvent to form hydrogen bonds. The X-ray crystal structure of one of the dibenzyl sulfide complexes has been determined, and the geometrical arrangement of the ligands has been determined by NOE NMR measurements at low temperature. The nature of the solvent, imine, sulfide, and halogen ligands produces differences in the reaction rates, which can be quantified very well by the corresponding DeltaS values that move from +48 to -90 J K(-1) mol(-1). The reaction on [Pt(Me)(2)F(C(5)CF(4)CHNCH(2)Ph) (SMe(2))] has been found to take place via a mechanism that depends strongly on the bulkiness of the substituting phosphine. While for PCy(3) the reaction is dissociative, for smaller entering ligands the first associatively activated substitution mechanisms on organometallic Pt(IV) complexes have been established with values of DeltaH and DeltaS in the 28-44 kJ mol(-1) and -120 to -83 J K(-1) mol(-1) ranges. Important intramolecular hydrogen bonding in the starting material can be held responsible for this difference with the remaining systems. PMID- 11925166 TI - Excited-state distortions determined from structured luminescence of nitridorhenium(V) complexes. AB - The nitridorhenium(V) complexes ReNCl(2)(PCy3)(2) (1), ReNBr(2)(PCy3)(2) (2), ReNCl(2)(PPh3)(2) (3), and ReNBr(2)(PPh3)(2) (4) produce structured emission spectra upon excitation at low temperature. The origin, E(00), occurs at 15 775, 16 375, 15 875, and 16 300 cm(-1), respectively. The vibronic peaks are regularly spaced with an average energy separation corresponding to the Re triple bond N stretching frequency. The nitridorhenium stretching frequency ranges from 1095 to 1101 cm(-1), as determined by Raman and IR spectroscopy. The excited-state distortions are calculated by fitting the emission spectra. The excited state arises primarily from a d(xy) (ReN nonbonding) to d(yz) (ReN pi antibonding) transition. The rhenium-nitrogen bond length in the excited state is 0.08 A longer than in the ground electronic state, which is consistent with the difference in bond lengths of ReN bonds of bond order 3 and bond order 2.5 as determined from molecular structures. PMID- 11925167 TI - Structure and bonding in [M(6)O(19)](n-) isopolyanions. AB - The structure and bonding in [M(6)O(19)](n-) isopolyanions of Nb, Ta, Mo, and W have been investigated using density-functional methods. The computational experimental agreement is good for the geometrical parameters of Mo and W species but less satisfactory for Nb and Ta clusters. The electronic structure of the anions has been probed with molecular-orbital, Mulliken-Mayer, and bonding-energy approaches. The results have indicated that M-O interactions are largely M d-O p in character and that sigma and pi bonds link the metal centers to terminal and bridging (O(b)) oxygen atoms. Some M-O(b) bonds exhibit a [M(4)O(4)] closed-loop structure, but this orbital-interaction mode has not been found to make a particularly outstanding contribution to the bonding stability of the molecules. Mayer indexes correspond to (fractional) multiple, approximately single, and low order character for terminal, bridging, and internal bonds, respectively, and the valency analysis has yielded similar bonding capacities for the different oxygen atoms. A distribution of the negative charge over all types of oxygen sites and metal charges considerably smaller than the formal oxidation states have been obtained from the Mulliken analysis. Minimal structural changes have been detected on reduction of molybdates and tungstates, in accord with the general properties of the orbitals occupied by the added electrons. PMID- 11925168 TI - Structural, spectral, and magnetic properties of end-to-end di-mu-thiocyanato bridged polymeric complexes of Ni(II) and Co(II). X-ray crystal structure of di mu-thiocyanatobis(imidazole)nickel(II). AB - Thiocyanatonickel(II) and thiocyanatocobalt(II) complexes of the composition Ni(NCS)(2)(HIm)(2) (1) and Co(NCS)(2)(HIm)(2) (2), where HIm = imidazole, were prepared and studied. In particular, the crystal structure of Ni(NCS)(2)(HIm)(2) was determined by X-ray methods. This compound crystallizes in the monoclinic system, space group P2(1)/n, with a = 7.720(1) A, b = 5.557(1) A, c = 13.774(3) A, beta = 102.54(3) degrees, and Z = 2. Its structure consists of a one dimensional polymeric chain in which nickel(II) ions are bridged by two thiocyanate groups bonding in an end-to-end fashion in a trans arrangement. The Ni...Ni distance is 5.557(1) A. The crystal packing is determined by the intermolecular hydrogen bonds and ring-stacking interactions. From their X-ray powder-diffraction patterns and IR spectra, the complexes 1 and 2 were found to be mutually isomorphous. The coordination compounds were identified and characterized using elemental analysis, magnetic measurements, and infrared and ligand-field spectra. Both complexes are first examples of ferromagnetically coupled one-dimensional polymeric compounds with double end-to-end thiocyanate bridges. The magnetic properties of the title compounds were investigated over the 1.9-290 K temperature range. The compounds exhibit long-range magnetic ordering with T(c) equal to 5.0 and 5.5 K for 1 and 2, respectively. Their isothermal magnetization was also studied. The magnetostructural properties of the nickel(II) compound obtained are discussed and compared to those of other double end-to-end thiocyanate-bridged nickel(II) complexes. PMID- 11925169 TI - Synthesis and structural characterization of integrally oxidized, metal-free phthalocyanine compounds: [H(2)(pc)][IBr(2)] and [H(2)(pc)](2)[IBr(2)]Br.C(10)H(7)Br. AB - The first integrally oxidized metal-free phthalocyanine compounds have been synthesized by chemical oxidation. Phthalocyanine (H(2)(pc), pc = phthalocyaninato) was oxidized with IBr to afford the compounds [H(2)(pc)][IBr(2)] (1) and [H(2)(pc)](2)[IBr(2)]BrAC(10)H(7)Br (2), whose structures were determined by means of single-crystal X-ray diffraction methods: [H(2)(pc)][IBr(2)], P2(1)/c, a = 8.0272(9) A, b = 21.258(2) A, c = 18.1439(2) A, beta = 113.975(2) degrees, V = 2828.8(5) A(3), T = 153 K, Z = 4; [H(2)(pc)](2)[IBr(2)]Br.C(10)H(7)Br, P, a = 8.4724(6) A, b = 13.5794(10) A, c = 13.8403(10) A, alpha = 90.854(1) degrees, beta = 103.417(1) degrees, gamma = 97.049(1)E degrees, V = 1535.61(19) A(3), T = 153 K, Z = 1. The extended structure of [H(2)(pc)][IBr(2)] comprises slipped columns of pc rings stacked along the a axis in adjacent columns at approximately 70 degrees to one another. IBr(2-) ions occupy the interstitial columns. The extended structure of [H(2)(pc)](2)[IBr(2)]Br.C(10)H(7)Br comprises slant stacks of pc rings along the crystallographic a axis with IBr(2-) ions, Br(-) ions, and disordered 1 bromonaphthalene molecules in the adjacent, parallel columns. The overall reaction for the formation of 1 is 2H(2)(pc) + 4IBr --> 2[H(2)(pc)][IBr(2)] + I(2), and the overall reaction for the formation of 2 (not including solvent) is 2H(2)(pc) + 3IBr --> [H(2)(pc)](2)Br[IBr(2)] + I(2). PMID- 11925170 TI - Pairwise exchanges of oxo and imido groups in rhenium(VII) compounds. AB - The kinetic and thermodynamic parameters for the oxo and imido exchange reactions among MeReO(3), MeReO(2)(NR), MeReO(NR)(2), and MeRe(NR)(3) (R = 1-adamantyl, Ad; or 2,6-diisopropylphenyl, Ar) have been measured. The rate constant for the NAr series decreases from 0.27 to 0.0024 L mol(-1) s(-1) at 25 degrees C in benzene as the total number of participating imido groups increases from 2 to 4, indicating that steric effects play an important role in the kinetics of the ligand exchange reactions. But, with NAd, the values of k/L mol(-1) s(-1) are 0.2 (4 NAd), 100 (3 NAd), and 0.74 (2 NAd). The equilibrium constants, also subject to steric effects, are much larger than those predicted by ligand combination statistics and greatly favor the mixed oxo-imido compounds. The different steric demands by imido and oxo ligands are believed to be the main factor for the larger equilibrium constants because the equilibrium constant shows minimal dependence on temperature. The large negative activation entropies for the ligand exchange reactions are consistent with a metathesis mechanism featuring nearly concurrent interchange of oxo and imido groups. PMID- 11925171 TI - Catecholase activity of a series of dicopper(II) complexes with variable Cu OH(phenol) moieties. AB - The catecholase activity of a series of dicopper(II) complexes containing different numbers of phenol groups coordinated to the metal centers was studied to identify functional as well as structural models for the type III copper enzymes tyrosinase and catechol oxidase. The syntheses and characterization of complexes [Cu(2)(H(2)bbppnol)(mu-OAc)(H(2)O)(2)]Cl(2).2H(2)O (1) and [Cu(2)(Hbtppnol)(mu-OAc)](ClO(4))(2) (2) were previously reported by us (Inorg. Chim. Acta 1998, 281, 111-115; Inorg. Chem. Commun. 1999, 2, 334-337), and complex [Cu(2)(P1-O(-))(OAc(-))](ClO(4))(2) (3) was previously reported by Karlin et al. (J. Am. Chem. Soc. 1997, 119, 2156-2162). The catalytic activity of the complexes 1-3 on the oxidation of 3,5-di-tert-butylcatechol was determined spectrophotometrically by monitoring the increase of the 3,5-di-tert-butyl-o benzoquinone characteristic absorption band at about 400 nm over time in methanol saturated with O(2)/aqueous buffer pH 8 solutions at 25 degrees C. The complexes were able to oxidize 3,5-di-tert-butylcatechol to the corresponding o-quinone with distinct catalytic activity. A kinetic treatment of the data based on the Michaelis-Menten approach was applied. The [Cu(2)(H(2)bbppnol)(mu OAc)(H(2)O)(2)]Cl(2) small middle dot2H(2)O complex showed the highest catalytic activity of the three complexes as a result of a high turnover rate (k(cat) = 28 h(-1)) combined with a moderate substrate-catalyst binding constant (K(ass) = 1.3 x 10(3) M(-1)). A mechanism for the oxidation reaction is proposed, and reactivity differences, k(cat)/K(M) of the complexes, were found to be dependent on (DeltaE)(1,2), the difference in the driving force for the reduction reactions Cu(II)(2)/Cu(II)Cu(I) and Cu(II)Cu(I)/Cu(I)(2). PMID- 11925172 TI - A dinuclear manganese(II) complex with the [Mn(2)(mu-O(2)CCH(3))(3)](+) core: synthesis, structure, characterization, electroinduced transformation, and catalase-like activity. AB - Reactions of Mn(II)(PF(6))(2) and Mn(II)(O(2)CCH(3))(2).4H(2)O with the tridentate facially capping ligand N,N-bis(2-pyridylmethyl)ethylamine (bpea) in ethanol solutions afforded the mononuclear [Mn(II)(bpea)](PF(6))(2) (1) and the new binuclear [Mn(2)(II,II)(mu-O(2)CCH(3))(3)(bpea)(2)](PF(6)) (2) manganese(II) compounds, respectively. Both 1 and 2 were characterized by X-ray crystallographic studies. Complex 1 crystallizes in the monoclinic system, space group P2(1)/n, with a = 11.9288(7) A, b = 22.5424(13) A, c =13.0773(7) A, alpha = 90 degrees, beta = 100.5780(10 degrees ), gamma = 90 degrees, and Z = 4. Crystals of complex 2 are orthorhombic, space group C222(1), with a = 12.5686(16) A, b = 14.4059(16) A, c = 22.515(3) A, alpha = 90 degrees, beta = 90 degrees, gamma = 90 degrees, and Z = 4. The three acetates bridge the two Mn(II) centers in a mu(1,3) syn-syn mode, with a Mn-Mn separation of 3.915 A. A detailed study of the electrochemical behavior of 1 and 2 in CH(3)CN medium has been made. Successive controlled potential oxidations at 0.6 and 0.9 V vs Ag/Ag(+) for a 10 mM solution of 2 allowed the selective and nearly quantitative formation of [Mn(III)(2)(mu O)(mu-O(2)CCH(3))(2)(bpea)(2)](2+) (3) and [Mn(IV)(2)(mu-O)(2)(mu O(2)CCH(3))(bpea)(2)](3+) (4), respectively. These results have shown that each substitution of an acetate group by an oxo group is induced by a two-electron oxidation of the corresponding dimanganese complexes. Similar transformations have been obtained if 2 is formed in situ either by direct mixing of Mn(2+) cations, bpea ligand, and CH(3)COO(-) anions with a 1:1:3 stoichiometry or by mixing of 1 and CH(3)COO(-) with a 1:1.5 stoichiometry. Associated electrochemical back-transformations were investigated. 2, 3, and the dimanganese [Mn(III)Mn(IV)(mu-O)(2)(mu-O(2)CCH(3))(bpea)(2)](2+) analogue (5) were also studied for their ability to disproportionate hydrogen peroxide. 2 is far more active compared to 3 and 5. The EPR monitoring of the catalase-like activity has shown that the same species are present in the reaction mixture albeit in slightly different proportions. 2 operates probably along a mechanism different from that of 3 and 5, and the formation of 3 competes with the disproportionation reaction catalyzed by 2. Indeed a solution of 2 exhibits the same activity as 3 for the disproportionation reaction of a second batch of H(2)O(2) indicating that 3 is formed in the course of the reaction. PMID- 11925173 TI - Carboxyester hydrolysis catalyzed by a novel dicopper(II) complex with an alcohol pendant macrocycle. AB - A novel hexaaza macrocycle bearing two hydroxyethyl pendants (L), 3,6,9,16,19,22 hexaaza-6,19-bis(2-hydroxyethyl)tricyclo[22,2,2,2(11,14)]triaconta 1,11,13,24,27,29-hexaene, was synthesized as a potential binucleating ligand. The corresponding Cu(II) complex [Cu(2)LCl(2)]Cl(2) small middle dot5.5H(2)O was isolated as a blue crystal, triclinic, space group P with a= 9.4920(19) A, b = 4.783(3) A, c = 16.553(3) A, alpha = 63.87(3) degrees, beta = 86.10(3) degrees, gamma = 83.8(3) degrees, V = 2072.8(7) A(-3), Z = 2, R1 = 0.0658, and wR2 = 0.1839. Both Cu ions adopt the geometry of a distorted trigonal bipyramid in a pentacoordinated environment. A complexation study on the novel title complex has revealed that the alcoholic OH groups of the complex Cu(2)L exhibit an obvious acidity with rather low pK(a) values at 25 degrees C. The Cu(II)-bound alkoxides, which act as reactive nucleophiles toward the hydrolysis of 4-nitrophenyl acetate in 10% (v/v) CH(3)CN at 25 degrees C, with I = 0.10 (NaNO(3)) and pH 9.3, have shown a second-order rate constant, 0.41 +/- 0.02 M(-1) s(-1), a value that is approximately 10 times greater than the corresponding value for the mononuclear Cu(II) complex formed by a relatively simple tripodal ligand (L1). The pH-rate profile gave a sigmoidal curve. The possible catalytic mechanism has been proposed, and the reason for the high catalytic activity of the title complex has been discussed. PMID- 11925174 TI - Magnetically isolated Cu(II)Gd(III) pairs in the series [Cu(acacen)Gd(pta)(3)], [Cu(acacen)Gd(hfa)(3)], [Cu(salen)Gd(pta)(3)], and [Cu(salen)Gd(hfa)(3)], [acacen = N,N'-ethylenebis(acetylacetoniminate(-)), salen = N,N' ethylenebis(salicylideniminate(-)), hfa = 1,1,1,5,5,5-hexafluoropentane-2,4 dionate(-), pta = 1,1,1-trifluoro-5,5-dimethylhexane-2,4-dionate(-)]. AB - [Cu(salen)Gd(pta)(3)] (1), [Cu(acacen)Gd(pta)(3)] (2), and [Cu(acacen)Gd(hfa)(3)] (3) are three heterobimetallic [Cu(II)Gd(III)] complexes of general formula [Cu(SB)Gd(beta-dik)(3)], in which a N,N',O,O' Schiff base (SB) ligand [acacen = N,N'-ethylenebis(acetylacetoniminate(-)), salen = N,N' ethylenebis(salicylideneiminate(-))] tetracoordinates Cu(II) and chelates Gd(III) as a tris(beta-diketonate) complex [hfa = 1,1,1,5,5,5-hexafluoropentane-2,4 dionate(-); pta = 1,1,1-trifluoro-5,5-dimethylhexane-2,4-dionate(-)]. They crystallize as a triclinic structure (space group P). The cell parameters are a = 9.8616(10) A, b = 12.1976(13) A, c = 18.4187(22) A, alpha = 90.671(14) degrees, beta = 100.588(13) degrees, gamma = 103.684(12) degrees, V = 2113 A(3), and Z = 2 for 1; a = 9.7560(11) A, b = 12.2924(13) A, c = 18.9368(22) A, alpha = 88.449(14) degrees, beta = 87.269(14) degrees, gamma = 67.629(12) degrees, V = 2098 A(3), and Z = 2 for 2; and a = 12.5726(15) A, b = 15.5985(18) A, c = 18.3724(21) A, alpha = 85.963(13) degrees, beta = 85.411(14) degrees, gamma = 80.766(14) degrees, V = 3539 A(3), and Z = 4 for 3. The Cu(O,O')Gd bridging cores show folding angles about O,O' in the range 139 degrees -147 degrees and intramolecular Cu small middle dot small middle dot small middle dotGd distances of about 3.3 A. In the solid state, the molecules form centrosymmetric pseudodimers [Cu(SB)Gd(beta-dik)(3)](2), through the overlap of the Cu(SB) entities. Resulting intradimer Cu...Cu distances are 5.941(1) A for 1, 4.831(1) A for 2, and 4.511(1) and 3.868(1) A for 3 which comprises two symmetrically independent dimers. The temperature dependence of complexes 1-3 was investigated in the range 1.8-300 K and revealed weak ferromagnetic interactions. Results are discussed in light of the structural features and of available magnetostructural data for other heterobimetallic [Cu(II)Gd(III)] complexes, including [Cu(salen)Gd(hfa)(3)] (4) (Ramade, I.; Kahn, O.; Jeannin, Y.; Robert, F. Inorg. Chem. 1997, 36, 930-936). PMID- 11925175 TI - N-centered hexazirconium chloride clusters: excision and redox chemistry. AB - The tightly cross-linked solid Zr(6)Cl(15)N yields [(Zr(6)NCl(12))Cl(6)](3-) upon heating with bis(triphenylphosphine)iminium chloride (PPNCl), in MeCN at 90 degrees C. Purple solutions containing [(Zr(6)NCl(12))Cl(6)](3-) were obtained and characterized with (15)N NMR. Cyclic voltammetric (CV) measurements on the series of [(Zr(6)ZCl(12))Cl(6)](n-) cluster ions (Z = Be, B, C, and N) in acetonitrile reveal that these cluster ions exhibit multiple reversible redox waves at potentials that can be systematically understood, including a reversible redox wave corresponding to the [(Zr(6)NCl(12))Cl(6)](3-/4-) couple. Preparation of the reduced cluster ion, [(Zr(6)NCl(12))Cl(6)](4-), (with 15 cluster-bonding electrons) was achieved by reduction of [(Zr(6)NCl(12))Cl(6)](3-) with (C(5)(CH(3))(5))(2)Co. Several new N-centered cluster complexes: (PPN)(3)[(Zr(6)NCl(12))Cl(6)].CH(2)Cl(2), [(C(5)(CH(3))(5))(2)Co(+)](3)[(Zr(6)NCl(12))Cl(6)], and (Et(4)N)(4)[(Zr(6)NCl(12))Cl(6)].2CH(3)CN have been isolated and structurally characterized. PMID- 11925176 TI - Effect of asymmetry on the electronic delocalization in diiron and iron-cobalt mixed valence metallocenic compounds. AB - In this work, we report the synthesis and a study on the degree of electronic delocalization in the asymmetric mixed valence complexes [CpFe(C(8)H(6))Fe(C(8)H(7))](+), 3a(+), and [CpCo(C(8)H(6))Fe(C(8)H(7))](+), 3b(+), (Cp = C(5)Me(5), C(8)H(6) = pentalenyde, C(8)H(7) = hydropentalenyde, and = ((3,5(CF(3))(2)C(6)H(3))(4)B(-))). Electrochemical methods, (57)Fe Mossbauer spectroscopy, electronic spectroscopy, and electron paramagnetic resonance were used for this purpose. Although the anti conformation of the complexes precludes any metal-metal interaction, all the techniques employed show that 3a(+) is a electronic delocalized system, while 3b(+) behaves as two individual metallic centers with localized electron density. PMID- 11925177 TI - Unique reactivity of a tetradentate N(2)S(2) complex of nickel: intermediates in the production of sulfur oxygenates. AB - Complex 1 [(N,N'-dimethyl-N,N'-bis(2-sulfanylethyl)ethylenediamine)nickel(II)], previously shown to react with H(2)O(2) to produce the fully oxygenated disulfonate 5 [diaqua(N,N'-dimethyl-N,N'-bis(2 sulfonatoethyl)ethylenediamine)nickel(II)], has been explored in detail to explain the observed reactivity of this compound and to discern intermediates in the oxygenation reaction. Reaction of 1 with 1 equiv of methyl iodide results in the monomethylated square-planar nickel complex 2 [[(N,N'-dimethyl-N-(2 sulfanylethyl)-N'-(2-methylthioethyl)(ethylenediamine)nickel(II)] iodide], while a slight excess of methyl iodide results in the dimethylated complex 3 [diiodo(N,N'-dimethyl-N,N'-bis(2-methylthioethyl)ethylenediamine)nickel(II)], an X-ray structure of which has shown that the nickel ion is in an octahedral N(2)S(2)I(2) environment. Crystal data of 3: monoclinic, a = 8.865(3) A, b = 14.419(4) A, c = 14.389(6) A, beta = 100.19(3) degrees, V = 1810.2(12) A(3), space group P2(1)/n, Z = 4. The equatorial positions are occupied by the two cis amine N-atoms and the coordinated iodides, while the axial positions are occupied by the thioether sulfur atoms. In organic solvents, the dithiolate complex 1 reacts with molecular oxygen or H(2)O(2) to produce the mixed sulfinato/thiolato complex 4 [(N,N'-dimethyl-N-(2-sulfanylethyl)-N'-(2 sulfinatoethyl)(ethylenediamine)nickel(II)], and the fully oxidized product 5. X ray analysis of complex 4 reveals a square-planar geometry in which the nickel ion is coordinated by two cis-amine nitrogens, one thiolate sulfur donor, and one sulfinato sulfur donor. Crystal data of 4: orthorhombic, a = 11.659(2) A, b = 13.119(3) A, c = 16.869(3) A, V = 2580.2(9) A(3), space group Pbca, Z = 8. This complex is the only intermediate in the oxygenation reaction that could be isolated, and it is shown to be further reactive toward O(2) to yield the fully oxidized product 5. For a better understanding of the reactivity observed for 4, DFT calculations have been undertaken, which show a possible reaction path toward the fully oxidized product 5. PMID- 11925178 TI - Geometric hydration shells for anionic platinum group metal chloro complexes. AB - Solvation shells surrounding complex inorganic anions have not been extensively studied and are often mentioned with an amorphous picture in mind. We use a computational model previously validated against experimental results and ab initio quantum calculations (Lienke, A.; Klatt, G.; Robinson, D.; Koch, K. R.; Naidoo, K. J. Inorg.Chem. 2001, 40, 2352-2357) to investigate the nature of the hydration shells about simple platinum group metal chloro complexes ([PtCl(6)](2 ), [RhCl(6)](3-), [PtCl(4)](2-), and [PdCl(4)](2-)). Our simulations show that the hydration shells surrounding these complexes are symmetric and take on familiar geometric forms. We find that only the [RhCl(6)](3-) complex has a clearly defined second hydration shell while the [PtCl(6)](2-), [PtCl(4)](2-), and [PdCl(4)](2-) second hydration shells are more diffuse. PMID- 11925179 TI - Mild benzene-thermal route to GaP nanorods and nanospheres. AB - GaP nanorods and nanospheres were synthesized from a mild benzene-thermal route at 240 and 300 degrees C, respectively, using Na, P, and GaCl(3) as the starting materials. The structure of the products was identified as zinc blende phase by X ray powder diffraction (XRD). Transmission electron microscopy (TEM) images showed that, when the reaction temperature was 240 degrees C, the products were nanorods with widths of 20-40 nm and lengths of 200-500 nm and nanospheres with diameters of 20-40 nm. However, when the reaction temperature was increased to 300 degrees C, the products were only nanospheres, and the diameters increased to 40-60 nm. The reaction proceeded through a metallic gallium intermediate, and a solution-liquid-solid (SLS) mechanism was proposed for the one-dimensional growth. The products were also investigated by UV-vis absorption and X-ray photoelectron spectroscopy. PMID- 11925180 TI - New mononuclear, cyclic tetranuclear, and 1-D helical-chain Cu(II) complexes formed by metal-assisted hydrolysis of 3,6-di-2-pyridyl-1,2,4,5-tetrazine (DPTZ): crystal structures and magnetic properties. AB - The reactions of 3,6-di-2-pyridyl-1,2,4,5-tetrazine (DPTZ) with different Cu(II) salts generate two new ligands, 2,5-bis(2-pyridyl)-1,3,4-oxodiazole (L(1)) and N,N'-bis(alpha-hydroxyl-2-pyridyl)ketazine (H(2)L(2)), from the metal-assisted hydrolysis of DPTZ, and form three new complexes: a mononuclear complex [Cu(L(1))(2)(H(2)O)(2)] .2ClO(4) (1), a linear coordination polymer [Cu(L(1))(NO(3))(2)](8) (2), and a cyclic tetranuclear complex [Cu(4)(L(2))(2)(Im)(2)(NO(3))(4)(H(2)O)(2)] (3) (Im = imidazole). Crystal data for 1: space group P2(1)/n with a = 10.339(3) A, b = 10.974(2) A, c = 13.618(4) A, beta = 103.24(1) degrees, and Z = 2. Crystal data for 2: space group C2/c with a = 13.9299(14) A, b = 9.2275(9) A, c = 12.1865(13) A, beta = 111.248(2) degrees, and Z = 4. Crystal data for 3: space group P2(1)/n with a = 9.3422(14) A, b = 15.987(2) A, c = 13.963(2) A, beta = 108.587(3) degrees, and Z = 2. L(1) acts as a bidentate chelating ligand in 1 and as a bis-bidentate chelating ligand in 2 with the shortest intramolecular Cu...Cu distance of 6.093 A. L(2) is a hexadentate ligand to bridge four Cu(II) ions, forming an interesting neutral cyclic tetranuclear complex 3 with Cu...Cu distances varying from 4.484 to 9.370 A. The mechanism of the metal assisted hydrolysis of DPTZ is discussed in detail. Magnetic susceptibility measurements indicate that 2 shows weak ferromagnetic interaction (J = 2.85 cm(-1)) along the 1-D helical chain, and that 3 displays weak antiferromagnetic interaction (J = -1.19 cm(-1) for the N-N bridge) and ferromagnetic interaction (j = 0.11 cm(-1) for the O-C=N bridge) between the adjacent Cu(II) ions. PMID- 11925181 TI - Self-assembly of transition-metal-based macrocycles linked by photoisomerizable ligands: examples of photoinduced conversion of tetranuclear-dinuclear squares. AB - A series of hetero- and homometallic square complexes bridged by a photoactive 4,4'-azopyridine (AZP) or 1,2-bis(4-pyridyl)ethylene (BPE) ligand, cyclobis[[cis (dppf)M](mu-L)(2)(fac-Re(CO)(3)Br)](OTf)(4) (M = Pd, L = trans-AZP (5); M = Pt, L = trans-AZP (7); M = Pd, L = trans-BPE (8); M = Pt, L = trans-BPE (10)), cyclo[[cis-(dppf)M](mu-L)(2)(fac-Re(CO)(3)Br)](OTf)(2) (M = Pd, L = cis-AZP (6); M = Pd, L = cis-BPE (9)), [cis-(dppf)Pd(mu-trans-AZP)](4)(OTf)(8) (11), and [cis (dppf)Pd(mu-cis-AZP)](2)(OTf)(4) (12), where dppf is 1,1' bis(diphenylphosphino)ferrocene and OTf is trifluoromethanesulfonate anion, were prepared by thermodynamically driven self-assembly processes. The photophysical and photochemical properties of these complexes have been investigated, and all of them show a lack of luminescence in room temperature solution. Upon irradiation at 313 or 366 nm, Pd(II)-Re(I)-containing tetranuclear squares 5, 8, and 11 undergo photoisomerization and convert to their corresponding dinuclear complexes 6, 9, and 12, whereas Pt(II)-Re(I)-based squares 7 and 10 show only slow square disassembling processes. The tetranuclear squares can be fully recovered by heating the photoisomerized solution for several hours. PMID- 11925182 TI - Synthesis of tris- and tetrakis(pyrazol-1-yl)borate gold(III) complexes. Crystal structures of [Au[kappa(2)-N,N'-BH(Pz)(3)]Cl(2)] (pz = pyrazol-1-yl) and [Au[kappa(2)-N,N'-B(Pz)(4)](kappa(2)-C,N-C(6)H(4)CH(2)NMe(2)-2)]ClO(4).CHCl(3). AB - Na[BH(pz)(3)] and Na[AuCl(4)].2H(2)O react in water (1:1) to give [Au[kappa(2) N,N'-BH(pz)(3)]Cl(2)] (1) or, in the presence of NaClO(4) (2:1:1), the cationic complex [Au[kappa(2)-N,N'-BH(pz)(3)](2)]ClO(4) (2). The reactions of Na[B(pz)(4)] with the cyclometalated gold complexes [AuRCl(2)] and NaClO(4) (1:1:1) produce [Au[kappa(2)-N,N'-B(pz)(4)](R)]ClO(4) [R = kappa(2)-C,N-C(6)H(4)CH(2)NMe(2)-2 (3)] or [Au[kappa(2)-N,N'-B(pz)(4)](R)Cl] [R = C(6)H(3)(N=NC(6)H(4)Me-4')-2-Me-5 (4)], respectively, although 4 is better obtained in the absence of NaClO(4). The crystal structures of 1 and 3.CHCl(3) are reported. Both complexes display the gold center in square planar environments, two coordination sites being occupied by the chelating poly(pyrazolyl)borate ligands. PMID- 11925183 TI - Pentagon stability: cyclic delocalization of lone pairs through sigma conjugation and design of polycyclophosphanes. AB - The orbital-phase theory was applied to propose pentagon stability in a well defined manner. Cyclic delocalization of the lone pair electrons on the five membered ring atoms through the vicinal sigma bonds was shown to be favored by the orbital-phase properties. The pentagon stability was found to be outstanding in saturated phosphorus five-membered rings in the puckered conformation, and was substantiated by the negative strain energy of cyclopentaphosphane, P(5)H(5) (3). The relative increments of the remarkable increase in the strain energies of protonation on the different atoms in the most stable conformers supported the significance of the cyclic delocalization of the lone pairs. Pentagon stability led to the design of three novel polycyclic phosphanes, P(12)H(4) (18), P(13)H(3) (19), and P(14)H(2) (20), with low strain energies due to many puckered pentagon units in them. The low stability of the dodecahedron P(20) (22) was suggested by the high strain energy due to its planar pentagon units. The pentagon stability is less significant in the saturated nitrogen ring molecules due to the greater energy gap between the n and sigma orbitals. PMID- 11925184 TI - A DFT study of the electronic spectrum of the alpha-keggin anion [Co(II)W(12)O(40)](6-). AB - Multiplet splittings for several excited configurations of [Co(II)W(12)O(40)](6-) were calculated using DFT methods. In agreement with the experimental interpretation of the spectrum the calculations found that the first strong band corresponds to Co d-d transitions, but it is worth noting that superposed to these transitions there are charge transfer transitions from cobalt to tungsten. The calculations also showed the importance of Jahn-Teller distortions in the excited states. With the exception of the consequences derived from a smaller splitting of d cobalt orbitals the d-d spectrum of [CoCl(4)](2-) is similar to that of the more complex Keggin anion. Finally, the energy of the bielectronic transition (4)A(2) --> (4)T(1)(P) was estimated via an approximate procedure based on ligand field theory. PMID- 11925185 TI - Coordination chemistry of novel scorpionate ligands based on 3-cyclohexylpyrazole and 3-cyclohexyl-4-bromopyrazole. AB - The ligands [hydrotris(3-cyclohexylpyrazol-1-yl)borate, [Tp(Cy)](-), tetrakis(3 cyclohexylpyrazol-1-yl)borate, [pz(o)Tp(Cy)](-), and hydrotris(3-cyclohexyl-4 bromopyrazol-1-yl)borate, [Tp(Cy,4Br)](-) were synthesized and characterized as their Tl(I) derivatives. They were converted to a variety of tetrahedral LMX and octahedral LML' complexes, as well as to the dinuclear nickel carbonate complex [Ni(Tp(Cy))](2)(CO(3)), 4, and the compound Ni[Tp(Cy,4Br)][pz(Cy,4Br)](3)(H)(2), 5. The structures of Co[Tp(Cy)]Cl, 1, Co[Tp(Cy,4Br)]Cl, 2, Co[Tp(Cy,4Br)]NCS, 3, [Ni(Tp(Cy))](2)(CO(3)), 4, Ni[Tp(Cy,4Br)][pz(Cy,4Br)](3)(H)(2), 5, and Mo[Tp(Cy)](CO)(2)(eta(3)-methallyl), 6, were determined by X-ray crystallography. The structures of paramagnetic heteroleptic complexes Co[Tp(Cy)][Tp], Co[Tp(Cy)][Tp], Co[Tp(Cy,4Br)][Tp], and Co[Tp(Cy,4Br)][Tp] were established by NMR. The homoleptic compounds Co[Tp(Cy)](2) and Co[Tp(Cy,4Br)](2) rearrange thermally to Co[Tp(Cy)](2) and to Co[Tp((Cy,4Br))](2), respectively, containing one 5-cyclohexyl group/ligand. PMID- 11925186 TI - Synthesis and resolution of (R,R)-(+/-)-1,1,4,7,10,10-hexaphenyl-1,10-diarsa-4,7 diphosphadecane: new ligand for the stereoselective self-assembly of dicopper(I), disilver(I), and digold(I) helicates. AB - Diphenylvinylarsine oxide reacts with 1,2-bis(phenylphosphino)ethane in the presence of potassium tert-butoxide to give the anti-Markovnikov product (R,R) (+/-)/(R,S)-1,1,4,7,10,10-hexaphenyl-1,10-diarsa-4,7-diphosphadecane dioxide 1AsO,10AsO, which, upon reduction with HSiCl(3)/NEt(3) in boiling acetonitrile, affords in 84% overall yield the di(tertiary arsine)-di(tertiary phosphine) (R,R) (+/-)/(R,S)-diphars. After separation of the diastereomers by fractional crystallization, the (R,R)-(+/-) form of the ligand was resolved by metal complexation with (+)-di(mu-chloro)bis[(R)-1-[1-(dimethylamino)ethyl]-2-phenyl C(2),N]dipalladium(II): (R,R)-diphars, mp 87-88 degrees C, has [alpha](D)(21) = 18.6 (c 1.0, CH(2)Cl(2)); (S,S)-diphars has [alpha](D)(21) = +18.4 (c 1.0, CH(2)Cl(2)). The crystal and molecular structures of the complexes (M) [M(2)[(R,R)-diphars](2)](PF(6))(2) (M = Cu, Ag, Au) have been determined: [M (S(Cu),S(Cu))]-(-)-[Cu(2)[(R,R)-diphars](2)](PF(6))(2), orthorhombic, P2(1)2(1)2(1) (No. 19), a = 16.084(3) A, b = 18.376(3) A, c = 29.149(6) A, Z = 4; [M-(S(Ag),S(Ag))]-(+)-[Ag(2)[(R,R)-diphars](2)](PF(6))(2), triclinic, P1, a = 12.487(2) A, b = 12.695(4) A, c = 27.243(4) A, alpha = 92.06 degrees, beta = 95.19 degrees, gamma = 98.23 degrees, Z = 2; [M-(S(Au),S(Au))]-(-)-[Au(2)[(R,R) diphars](2)](PF(6))(2), orthorhombic, P2(1)2(1)2(1) (No. 19), a = 16.199(4) A, b = 18.373(4) A, c = 29.347(2) A, Z = 4. In the copper(I) and gold(I) helicates, each ligand strand completes 1.5 turns of an M helix in a parallel arrangement about the two chiral MAs(2)P(2) stereocenters of S configuration. The unit cell of the silver(I) complex contains one molecule each of the parallel helicate of M configuration and the conformationally related double alpha-helix of M configuration in which each ligand strand completes 0.5 turns of an M helix about two metal stereocenters of S configuration. Energy minimization calculations of the three structures with use of the program SPARTAN 5.0 gave results that were in close agreement with the core structures observed. PMID- 11925187 TI - The synthesis and characterization of compounds of the type Hg[1-C(6)H(4)-2 C(H)=NC(6)H(5-n)R(n)](2). AB - The organomercurial compounds Hg[1-C(6)H(4)-2-C(H)=NC(6)H(5-n)R(n)](2) (R = 4 NMe(2), 6a; 4-Me, 6b; 4-I, 6c; 4-NO(2), 6d; 2-(i)Pr, 6e; 2-Me, 6f; 2,6-(i)Pr(2), 6g; 2,6-Me(2), 6h) have been prepared in good overall yield from 2 bromobenzaldehyde. All of the compounds have been characterized by elemental analysis, (1)H NMR, (13)C[(1)H] NMR, and infrared spectroscopy. In addition, compounds 6a [C(30)H(30)HgN(4), triclinic, P, a = 6.20000(10) A, b = 9.2315(2) A, c = 10.9069(3) A, alpha = 85.8510(10) degrees, beta = 89.3570(10) degrees, gamma = 87.206(2) degrees, Z = 1], 6b [C(28)H(24)HgN(2), monoclinic, P2(1)/c, a = 12.8260(5) A, b = 14.0675(4) A, c = 6.1032(2) A, beta = 90.0990(10) degrees, Z = 2], 6g [C(38)H(44)HgN(2), triclinic, P, a = 8.2626(2) A, b = 9.8317(2) A, c = 11.8873(3) A, alpha = 103.6650(10) degrees, beta = 109.3350(10) degrees, gamma = 104.627(2) degrees, Z = 1], and 6h [C(30)H(28)HgN(2), monoclinic, P2(1)/c, a = 12.5307(2) A, b = 10.9852(2) A, c = 18.2112(2) A, beta = 104.0190(10) degrees, gamma = 87.206(2) degrees, Z = 4] have been characterized by low-temperature single-crystal X-ray diffraction studies, and two different molecular geometries about the central mercury atom have been observed; intramolecular contacts suggest a van der Waals radius for Hg of 2.1-2.2 A. PMID- 11925188 TI - Layered zirconium phosphate chloride dimethyl sulfoxide as a two-dimensional exchanger of anionic ligands. Part I. Substitution of chloride with inorganic monodentate ligands. AB - Crystalline ZrPO(4)Cl(CH(3))(2)SO was prepared by direct precipitation in the presence of oxalic acid as a zirconium complexing agent. The structure of ZrPO(4)Cl(CH(3))(2)SO, refined with the Rietveld method using X-ray powder diffraction data, was confirmed to be close to that of the compound prepared using gamma-zirconium phosphate as a precursor. Chloride anions directly bonded to zirconium were found to act as weak ligands; this made possible their replacement with other monodentate anionic ligands. The preparation and a preliminary characterization of a series of inorganic derivatives obtained by topotactic replacement of Cl with OH, Br, MSO(4) (M = H, NH(4), Na), NaMoO(4), and HCrO(4) anions is reported. The possibility of replacement of chloride also with organic anions, such as alkoxides and carboxylates, and the possibility of substituting also dimethyl sulfoxide with other neutral ligands, as shown by preliminary study, makes ZrPO(4)Cl(CH(3))(2)SO a useful and very flexible precursor for materials chemistry. PMID- 11925189 TI - Effect of metal-oxygen covalent bonding on the competition between Jahn-Teller distortion and charge disproportionation in the perovskites of high-spin d(4) metal ions LaMnO(3) and CaFeO(3). AB - The perovskites LaMnO(3) and CaFeO(3) consisting of high-spin d(4) transition metal ions undergo different types of distortions, i.e., a Jahn-Teller distortion in LaMnO(3) and a charge disproportionation in CaFeO(3). We investigated the electronic factor causing this difference on the basis of first principles spin polarized electronic band structure calculations for their ideal cubic structures and also tight-binding electronic band structure calculations for their ideal cubic and distorted structures. Our study shows that a charge disproportionation is favored over a Jahn-Teller distortion in CaFeO(3) because the covalent character is strong in the Fe-O bond, while the opposite is true for LaMnO(3) because the covalent character is weak in the Mn-O bond. In spin-polarized electronic band structure calculations, the covalency of the M-O (M = Fe, Mn) bond is enhanced in the up-spin bands but is reduced in the down-spin bands. Our analysis shows that electron-electron repulsion causes the energy gap between the metal 3d and the oxygen 2p orbitals to become larger for the down-spin than for the up-spin-orbital interactions. Thus in the d-block e(g) bands of both LaMnO(3) and CaFeO(3) the metal 3d orbital contribution is larger in the down-spin than in the up-spin bands. PMID- 11925190 TI - Metallonitrosyl fragment as electron acceptor: intramolecular charge transfer, long range electronic coupling, and electrophilic reactivity in the trans [NCRu(py)(4)(CN)Ru(py)(4)NO](3+) ion. AB - The new complex trans-[NCRu(py)(4)(CN)Ru(py)(4)NO](PF(6))(3) (I) was synthesized. In acetonitrile solution, I shows an intense visible band (555 nm, epsilon = 5800 M(-1) cm(-1)) and other absorptions below 350 nm, associated with d(pi) --> pi(py) and pi(py) --> pi(py) transitions. The visible band is presently assigned as a donor-acceptor charge transfer (DACT) transition from the remote Ru(II) to the delocalized [Ru(II)-NO(+)] moiety. Photoinduced release of NO is observed upon irradiation at the DACT band. Application of the Hush model reveals strong electronic coupling, with H(DA) = approximately 2000 cm(-1). The difference between the optical absorption energy and redox potentials for the donor and acceptor sites (Ru(III,II), 1.40 V, and NO(+)/NO, 0.50 V, vs Ag/AgCl, 3 M KCl, respectively) (hnu - DeltaE(red)) is 1.33 eV, a large value which probably relates to the significant changes in distances and angles for the Ru-N-O moiety upon reduction. UV-vis absorptions, IR frequencies, and redox potentials are solvent-dependent. Controlled potential reduction (of NO(+)) and oxidation (of Ru(II) associated with the dicyano-chromophore) of I afford stable species, [NCRu(II)(py)(4)(CN)Ru(py)(4)NO](2+) (I(red)) and [NCRu(III)(py)(4)(CN)Ru(py)(4)NO](4+) (I(ox)), respectively, which are characterized by UV-vis and IR spectroscopies. I(red) shows an EPR spectrum characteristic of [Ru(II)-NO(*)] complexes. Compound I is electrophilically reactive in aqueous solution above pH 5: values of the equilibrium constant for the reaction [NCRu(py)(4)(CN)Ru(py)(4)NO](3+)+ 2 OH(-) <--> [NCRu(py)(4)(CN)Ru(py)(4)NO(2)](+) + H(2)O, K = 3.2 +/- 1.4 x 10(15) M(-2), and of the rate constant for the nucleophilic addition of OH(-), k = 9.2 +/- 0.2 x 10(3) M(-1) s(-1)(25 degrees C, I = 1 M), are obtained, with DeltaH = 90.7 +/- 3.8 kJ mol(-1) and DeltaS = 135 +/- 13 J K(-1) mol(-1). The oxidized complex, I(ox), shows an enhanced electrophilic reactivity toward OH(-). This addition reaction is followed by irreversible processes, which most probably lead to disproportionation of bound nitrite and other products. PMID- 11925191 TI - Synthesis and structural comparison of triaryl(sulfonylimino)pnictoranes. AB - Triarylphosphanes 1 (Ar(3)P; Ar = Ph, 4-MeC(6)H(4)), triphenylarsane (2), and triarylstibanes 3 (Ar(3)Sb; Ar = 2-MeC(6)H(4), 2-MeOC(6)H(4)) reacted with trifluoromethanesulfonamide (7a) in the presence of equimolar diethyl azodicarboxylate to afford the corresponding triaryl(sulfonylimino)pnictoranes [Ar(3)M=NSO(2)CF(3); 8 (M = P), 9 (M = As), 10 (M = Sb)]. The Kirsanov-type reaction of triarylantimony dichlorides 5 (Ar(3)SbCl(2); Ar = 2-MeC(6)H(4), 2 MeOC(6)H(4)) and triarylbismuth dichlorides 6 (Ar(3)BiCl(2); Ar = 2-MeC(6)H(4), 2 MeOC(6)H(4), 2,4,6-Me(3)C(6)H(2)) with sulfonamides 7 (H(2)NSO(2)R; R = CF(3), 4 MeC(6)H(4), Me) in the presence of 2 equiv of potassium tert-butoxide yielded triaryl(sulfonylimino)-lambda(5)-stibanes 10 and -bismuthanes 11, respectively. The ortho-substitution in aryl ligands of 10 and 11 has been found to bring about considerable kinetic stabilization of the reactive Sb=N and Bi=N bonds. A structural comparison was made for a series of triaryl(sulfonylimino)pnictoranes 8-11 by IR spectroscopy and X-ray crystallography. In the IR spectra of 9-11, SO(2) asymmetric stretching absorptions (nu(SO2)) were observed at lower wavenumbers as compared to those of phosphorus counterparts 8. The difference in frequency (Deltanu(SO2)) from 8 increased progressively as the pnictogen element being utilized moved down the group 15 column on the periodic table. X-ray crystallographic analyses of eight of the triaryl(sulfonylimino)pnictoranes prepared confirmed the increasing single-bond character of the M=N bond, with the contribution from the canonical structure M(+)-N=S(O)-O(-) increasing in importance in the order P < As < Sb < Bi. Among all triaryl(sulfonylimino)pnictoranes examined, only imino-lambda(5)-bismuthanes 11 oxidized alcohols to carbonyl compounds. PMID- 11925192 TI - Lowering melting points in asymmetrically substituted Salen-copper(II) complexes exhibiting mesomorphic behavior. Structure of the mesogen Cu(5-hexyloxySalen). AB - In comparison with their symmetrical analogues, unsymmetrically substituted Cu Salen complexes show mesophases with lowered melting points. For terminally substituted complexes, symmetrical ones (R(1) = R(2)) have only an S(A) phase, while for unsymmetrical alkoxy substitution a monotropic S(E) phase occurs and the melting temperature decreases with no loss in mesophase stability. Lateral substitution, when it is symmetrical (R(3) = R(4)), lowers mesophase stability but not melting temperature, and when it is unsymmetrical, it greatly lowers both mesophase stability and melting temperature compared with the parent compound. Substitution at the imine carbon (R(5), R(6)) also lowers chemical stability (decomposition) of the compounds. The structure of the 5-hexyloxy complex (R(1) = R(2) = OC(6)H(13), R(3) = R(4) = R(5) = H) shows the pre-mesophasic arrangement likely adopted after melting. PMID- 11925193 TI - Oxygen transfer reactions. 4. Reaction of high valent oxoruthenium compounds with sulfides. AB - The oxidation of methoxy substituted benzyl phenyl sulfides can be used to distinguish between oxidants that react by single electron transfer (followed by oxygen rebound) and those which react by direct oxygen atom transfer in a two electron process. Transfer of a single electron results in the formation of an intermediate radical cation, which can undergo C-S bond cleavage and deprotonation reactions leading to the formation of methoxy substituted benzyl derivatives, methoxy substituted benzaldehydes, and diphenyl disulfide. The oxidation of 4-methoxybenzyl phenyl sulfide and 3,4,5-trimethoxybenzyl phenyl sulfide by oxidants known to participate in single electron transfers (Ce(4+), Mn(3+), and Cr(6+)) results in the formation of the corresponding benzaldehydes, benzyl alcohols, benzyl acetates, and benzyl nitrates in variable yields. However, the only products obtained from the oxidation of the same compounds with RuO(4), RuO(4-), and RuO(4)(2-) are sulfoxides and sulfones. Therefore, it is concluded that the oxidation of sulfides by oxoruthenium compounds likely proceeds by a concerted mechanism. PMID- 11925194 TI - Redox chemistry of tellurium bis(tert-butylamido)cyclodiphosph(V)azane disulfide and diselenide systems: a spectroscopic and structural study. AB - The redox chemistry of tellurium-chalcogenide systems is examined via reactions of tellurium(IV) tetrachloride with Li[(t)()BuN(E)P(mu-N(t)Bu)(2)P(E)N(H)(t)Bu] (3a, E = S; 3b, E = Se). Reaction of TeCl(4) with 2 equiv of 3a in THF generates the tellurium(IV) species TeCl(3)[HcddS(2)][H(2)cddS(2)] 4a [cddS(2) = (t)BuN(S)P(mu-N(t)Bu)(2)P(S)N(t)Bu] at short reaction times, while reduction to the tellurium(II) complex TeCl(2)[H(2)cddS(2)](2) 5a is observed at longer reaction times. The analogous reaction of TeCl(4) and 3b yields only the tellurium(II) complex TeCl(2)[H(2)cddSe(2)](2) 5b. The use of 4 equiv of 3a or 3b produces Te[HcddE(2)](2) (6a (E = S) or 6b (E = Se)). NMR and EPR studies of the 5:1 reaction of 3a and TeCl(4) in THF or C(6)D(6) indicate that the formation of the Te(II) complex 6a via decomposition of a Te(IV) precursor occurs via a radical process to generate H(2)cddS(2). Abstraction of hydrogen from THF solvent is proposed to account for the formation of 2a. These results are discussed in the context of known tellurium-sulfur and tellurium-nitrogen redox systems. The X ray crystal structures of 4a.[C(7)H(8)](0.5), 5a, 5b, 6a.[C(6)H(14)](0.5), and 6b.[C(6)H(14)](0.5) have been determined. The cyclodiphosph(V)azane dichalcogenide ligand chelates the tellurium center in an E,N (E = S, Se) manner in 4a.[C(7)H(8)](0.5), 6a.[C(6)H(14)](0.5), and 6b.[C(6)H(14)](0.5) with long Te N bond distances in each case. Further, a neutral H(2)cddS(2) ligand weakly coordinates the tellurium center in 4a small middle dot[C(7)H(8)](0.5) via a single chalcogen atom. A similar monodentate interaction of two neutral ligands with a TeCl(2) unit is observed in the case of 5a and 5b, giving a trans square planar arrangement at tellurium. PMID- 11925196 TI - Third-order nonlinear optical properties of complexes with MM triple and quadruple bonds (M = Mo, W) at 1064 nm by degenerate four-wave mixing. AB - Measurements of the third-order nonlinear optical responses of solutions of the metal-metal multiply bonded complexes Mo(2)(OPr(i))(6), W(2)(OBu(t))(6), M(2)(NMe(2))(6), M(2)(O(2)CBu(t))(4), and M(2)Cl(4)(PMe(3))(4) (M = Mo, W), using picosecond degenerate four-wave mixing at 1064 nm, are reported. These complexes display only very small instantaneous electronic polarizations when excited with cross-polarized beams. When the excitation beams are similarly polarized, a significant third-order optical response is detected, which is attributable to the formation of bulk thermal excitation gratings. Time-dependent measurements support this view. PMID- 11925195 TI - Oxidative damage by ruthenium complexes containing the dipyridophenazine ligand or its derivatives: a focus on intercalation. AB - Interactions with DNA by a family of ruthenium(II) complexes bearing the dppz (dppz = dipyridophenazine) ligand or its derivatives have been examined. The complexes include Ru(bpy)(2)(dppx)(2+) (dppx = 7,8-dimethyldipyridophenazine), Ru(bpy)(2)(dpq)(2+) (dpq = dipyridoquinoxaline), and Ru(bpy)(2)(dpqC)(2+) (dpqC = dipyrido-6,7,8,9-tetrahydrophenazine). Their ground and excited state oxidation/reduction potentials have been determined using cyclic voltammetry and fluorescence spectroscopy. An intercalative binding mode has been established on the basis of luminescence enhancements in the presence of DNA, excited state quenching, fluorescence polarization values, and enantioselectivity. Oxidative damage to DNA by these complexes using the flash/quench method has been examined. A direct correlation between the amount of guanine oxidation obtained via DNA charge transport and the strength of intercalative binding was observed. Oxidative damage to DNA through DNA-mediated charge transport was also compared directly for two DNA-tethered ruthenium complexes. One contains the dppz ligand that binds avidly by intercalation, and the other contains only bpy ligands, that, while bound covalently, can only associate with the base pairs through groove binding. Long range oxidative damage was observed only with the tethered, intercalating complex. These results, taken together, all support the importance of close association and intercalation for DNA-mediated charge transport. Electronic access to the DNA base pairs, provided by intercalation of the oxidant, is a prerequisite for efficient charge transport through the DNA pi stack. PMID- 11925197 TI - Five-coordinate iron(III) porphycenes: (1)H NMR, magnetic, and structural studies. AB - Five-coordinate iron(III) 2,7,12,17-tetrapropylporphycene (TPrPc)Fe(III)X (X = C(6)H(5)O(-), Cl(-), Br(-), I(-), ClO(4)(-)) complexes have been investigated. The (1)H NMR spectra demonstrate downfield shifts for pyrrole resonances [(TPrPc)Fe(III)(C(6)H(5)O), 65.3 ppm; (TPrPc)Fe(III)Cl, 28.5 ppm] but large upfield ones for (TPrPc)Fe(III)Br (-7.8 ppm), (TPrPc)Fe(III)I (-49.4 ppm), and (TPrPc)Fe(III)ClO(4) (-77.1 ppm) (294 K, CD(2)Cl(2)). The pyrrole chemical shifts span the remarkable +70 to -80 ppm range. The variable-temperature (1)H NMR spectra of (TPrPc)Fe(III)X demonstrate anti-Curie behavior with a sign reversal for (TPrPc)Fe(III)Cl. These behaviors are consistent with the admixed S = 3/2, 5/2 ground electronic state with a dominating contribution of the S = 3/2 one. In terms of the chemical shift, (TPrPc)Fe(III)(ClO(4)) can be considered as an example of the purest S = 3/2 state in the investigated series. The extent of the S = 5/2 contribution in the admixed S = 3/2, 5/2 ground electronic state, as gradated solely the basis of the pyrrole proton paramagnetic shifts, is controlled by the strength of the axial ligand, following the magnetochemical series (Evans, D. R.; Reed, C. A. J. Am. Chem. Soc. 2000, 122, 4660). Significantly iron(III) 2,7,12,17-tetrapropylporphycene, soluble in typical organic solvents, can be considered as a universal framework to classify the ligand strength in a magnetochemical series, consistently using the beta-H pyrrole paramagnetic shifts as a fundamental criterion. The structure of (TPrPc)Fe(III)Cl has been determined by X-ray crystallography. The iron is five coordinate with bonds of nearly equal length to the four pyrrole nitrogen atoms (Fe-N in the range 1.983(5)-2.006(6) A). The iron lies 0.583(1) A out of the mean plane of the macrocycle and 0.502(5) A out of the mean N(4) plane. In the solid, pairs of molecules are positioned about the center of symmetry so there is face to-face pi-pi contact. The mean plane separation is 3.38 A, and the lateral shift of the porphycene center along the Fe-N bond is 4.490 A. The distance from one porphycene center to the other is 5.62 A, and the iron-iron separation is 6.304(2) A. PMID- 11925198 TI - New hydrogen bond-supported 3-D molecular assembly from polyoxovanadate and tetramethylbiimidazole. PMID- 11925200 TI - In situ formation of allyl ketones via Hiyama-Nozaki reactions followed by a chromium-mediated Oppenauer oxidation. AB - In Hiyama-Nozaki reactions of allylchromium with aldehydes the expected products are homoallylalcohols. However, oxidation products derived from these, predominantly allyl ketones, can be common side products. This can be explained by an Oppenauer-(Meerwein-Ponndorf-Verley)-type mechanism (OMPV-reaction). The amount of oxidation is strongly dependent on the substitution pattern of the reaction partners and the reaction conditions. An appropriate choice of these can lead to preferential formation of ketones instead of the alcohols. In addition to its synthetic usefulness, the oxidation-reduction equilibrium is of the utmost importance for the design of enantioselective Hiyama-Nozaki reactions because it is also a potential racemization pathway. PMID- 11925201 TI - Enantioselective allyltitanations and metathesis reactions. Application to the synthesis of piperidine alkaloids (+)-sedamine and (-)-prosophylline. AB - An enantioselective synthesis of the piperidine alkaloids (+)-sedamine and (-) prosophylline is reported. The synthesis of (+)-sedamine has been achieved in 12 steps with an overall yield of 20% from benzaldehyde, and (-)-prosophylline was obtained in 15 steps with an overall yield of 9.2%, starting from D glyceraldehyde acetonide 14. The key steps are enantioselective allyltitanation reactions and ring-closing or cross-metathesis reactions. PMID- 11925203 TI - Dications of fluorenylidenes. Relationship between electrochemical oxidation potentials and antiaromaticity in diphenyl-substituted fluorenyl cations. AB - The antiaromaticity of a series of dications of p-substituted diphenylmethylidene fluorenes was explored using three criteria attributed to aromaticity/antiaromaticity. The relative stability of the dications (energetic criterion) was measured via the redox potentials obtained by electrochemical oxidation under very fast sweep rates with microelectrodes. Comparison of redox potentials with those of a model system, p-substituted tetraphenylethylenes, shows relatively small destabilization of the potentially antiaromatic fluorenylidene dication. However, the amount of destabilization is comparable with the limited electrochemical data available for other antiaromatic systems. Nucleus independent chemical shifts (NICS) were calculated for these dications (magnetic criterion) and indicated their antiaromaticity. A good linear relationship between experimental and calculated (B3LYP/6-31G(d)) (1)H and (13)C NMR shifts for the three dications, 3c, 3e, and 3f, for which NMR data has been reported, validated the accuracy of the NICS values. Bond length alternation/elongation (structural criterion) was explored via the harmonic oscillator model of aromaticity (HOMA) using the geometries calculated with density functional theory, but there was insufficient variation to evaluate relative antiaromaticity. In addition, the presence of benzannulation appears to restrict bond length alternation to such an extent that the magnitude of the HOMA index is of little use in evaluating the antiaromaticity of many polycyclic hydrocarbons. Both NICS values and redox potentials for formation of the dication in these systems show a strong linear correlation with sigma(p)(+) values, with the more antiaromatic fluorenylidene dication possessing the more electron withdrawing substituent. The correlation between NICS values and redox potentials is also good, as might be expected, suggesting a strong relationship between magnetic and energetic characteristics of antiaromaticity. However, magnetic characteristics appear to be a more sensitive probe than energetic characteristics evaluated through redox potentials or structural characteristics evaluated through HOMA calculations. PMID- 11925202 TI - Effect of multivalency on the performance of enantioselective separation media for chiral HPLC prepared by linking multiple selectors to a porous polymer support via aliphatic dendrons. AB - Chiral stationary phases (CSPs) containing L-proline indananilide chiral selectors attached through a multivalent dendritic linker to monodisperse macroporous poly(2-hydroxyethyl methacrylate-co-ethylene dimethacrylate) beads have been prepared using two different approaches. The convergent method involves the preparation of ligands in solution and their subsequent attachment to the support. The divergent approach is based on the stepwise "on-bead" formation of the linker using methods that are typical of solid-phase synthesis. While the convergent CSPs feature well-defined ligands, their loading is relatively low. In contrast, the divergent technique affords CSPs with higher loading but with more limited control over precise ligand architecture. Excellent enantioselectivities characterized by separation factors of up to 31 were achieved for the separation of racemic N-(3,5-dinitrobenzoyl)-alpha-amino acid alkyl amides with these new CSPs under normal-phase HPLC conditions. PMID- 11925205 TI - Regiocontrolled synthesis of enantiopure 3,3'-thiosubstituted biphenyls. AB - Sulfenylation of 6,6'-dimethoxy-2,2'-dihydroxybiphenyl, used as a racemic mixture and single enantiomers, by phthalimidesulfenyl chloride afforded the corresponding 3,3'-N,N'-dithiophthalimide with complete regioselectivity. Simple manipulations of the latter compound allowed access to the corresponding bis thiol or o-thioquinone as useful intermediates for the synthesis of new sulfur containing open-chain and macrocyclic C(2) enantiopure ligands. The application of this methodology to the preparation of a biphenyl bearing two cysteine units as potential HIV-1 protease inhibitor is also described. PMID- 11925204 TI - Diastereoselective reactions in glycine templates containing an ent-ardeemin fragment. AB - Self-consistent reaction field solvation models derived from SCF-MO calculations are shown to be reliable in modeling the diastereoselectivity of the reactions of the anion and cation derived from (4S)-2,4-dimethyl-2,4-dihydro-1H-pyrazino[2,1 b]quinazoline-3,6-dione (1) at C(1) with electrophiles and nucleophiles, respectively. The found anti/syn ratio of compound 8, which is a seco-ent ardeemin analogue obtained by alkylation of 1 with gramine methiodide, confirms this computational model. A close similarity between the calculated geometry of the piperazine ring in the anti isomers of 1,2,4-trialkyl derivatives and that deduced from their (1)H NMR (solution) and X-ray data has been also established. PMID- 11925206 TI - Synthesis of 2-alkylidenecyclopentanones via palladium-catalyzed carbopalladation/ring expansion of 1-(1-alkynyl)cyclobutanols. AB - The palladium-catalyzed cross-coupling of aryl halides or vinylic halides or triflates and 1-(1-alkynyl)cyclobutanols affords good yields of stereoisomerically pure 2-arylidene- or 2-(2-alkenylidene)cyclopentanones, respectively. The process involves (1) oxidative addition of the organic halide or triflate to Pd(0), (2) regioselective, intermolecular carbopalladation of the carbon-carbon triple bond of the 1-(1-alkynyl)cyclobutanol to produce a vinylic palladium intermediate, (3) regioselective ring expansion to a palladacycle, and (4) reductive elimination of the 2-alkylidenecyclopentanone with simultaneous regeneration of the Pd(0) catalyst. Generally, the best results are obtained by employing 10 mol % of Pd(OAc)(2), 20 mol % of PPh(3), 2 equiv of the aryl or vinylic iodide or vinylic triflate, 2 equiv of diisopropylethylamine, and n Bu(4)NCl in DMF as the solvent. PMID- 11925207 TI - Isomerization of substituted biphenyls by superacid. A remarkable confluence of experiment and theory. AB - Acid-catalyzed isomerization of dimethylbiphenyls is determined by the relative stability of intermediate carbocations, rather than the neutral products, and may be predicted by a simple semiempirical method (AM1). A general kinetic model for such isomerizations is suggested in which the rearrangement of an intermediate cation is the rate-limiting step. Control of regiochemistry of dialkylbiphenyls provides a useful entry into high-purity monomers for high-polymer synthesis. PMID- 11925208 TI - Intramolecularly competitive Ireland-Claisen rearrangements: scope and potential applications to natural product synthesis. AB - A variety of bis-allylic esters were prepared by vinylmetal addition to cycloalkenones followed by esterification either in situ or in a separate operation. For chiral cyclohexenones, the vinyl additions generally occurred with >10:1 diastereoselectivity. Although in some cases the bis-allylic esters proved to be sensitive to silica gel or other adsorbents, all of the esters examined could be isolated in acceptable purity. The Ireland-Claisen rearrangement of the bis-allylic esters occurred with complete regioselectivity via the exocyclic alkene. The alkene stereochemistry and the stereochemistry at C-2 and C-3 of the pentenoic acid products were consistent with a chairlike transition state in the rearrangement. Substituents at the carbons adjacent to the allylic carbinol carbon (i.e., C-2 or C-6 in cyclohexenone-derived substrates) directed the stereochemical course of the rearrangement. The rearrangements generally proceeded so as to place the larger of the C-2 or C-6 substituents in the pseudoequatorial position with respect to the chairlike transition state. For a bis-allylic ester bearing both a C-2-CH(3) and a C-6-OMEM substituent, the rearrangement product resulted from the nominally smaller OMEM substituent occupying a pseudoequatorial position with respect to the chairlike transition state. PMID- 11925209 TI - Diastereoselective nucleophilic substitution reactions of oxasilacyclopentane acetals: application of the "inside attack" model for reactions of five-membered ring oxocarbenium ions. AB - The additions of nucleophiles to oxocarbenium ions derived from oxasilacyclopentane acetates proceeded with high diastereoselectivity in most cases. Sterically demanding nucleophiles such as the silyl enol ether of diethyl ketone add to the face opposite the C-2 substituent. These reactions establish the syn stereochemistry about the newly formed carbon-carbon bond. Small nucleophiles such as allyltrimethylsilane do not show this same stereochemical preference: they add from the same face as the substituent in C-2-substituted oxocarbenium ions. The stereoselectivities exhibited by both small and large nucleophiles can be understood by application of the "inside attack" model for five-membered ring oxocarbenium ions developed previously for tetrahydrofuran derived cations. This stereoelectronic model requires attack of the nucleophile from the face of the cation that provides the products in their lower energy staggered conformations. Small nucleophiles add to the "inside" of the lower energy ground-state conformer of the oxocarbenium ion. In contrast, sterically demanding nucleophiles add to the inside of the envelope conformer where approach is anti to the C-2 substituent of the oxocarbenium ion, regardless of the ground state conformer population. PMID- 11925210 TI - A novel crown ether generation containing different heteroaromatic cations: synthesis, characterization, solid-phase (13)C NMR, X-ray crystal structure, and selective amino acid recognition. AB - The synthesis and structural characterization of a novel generation of crown ethers, 3, 5 and 6 containing pyrilium, thiopyrilium, and pyridinium subunits, respectively, are reported. The crown ether unit is potentially capable of forming host-guest complexes with inorganic and organic cations, while the heteroaromatic cationic unit is suitable to bind with anions. A variety of physicochemical methods including electrospray mass spectrometry, UV-vis spectroscopy, solution and solid-phase NMR, and X-ray crystallography were applied for structural characterization of the new crown ether derivatives. The (1)H and (13)C NMR studies indicate rapid rotation of the B9C3 unit about the C-C bond that connects the two units to each other. Single crystals for 3, 4, and 5 were successfully obtained, and their X-ray crystal structures were resolved. The perchlorate anion in 3 (orthorhombic, space group P2(1)2(1)2(1)) and 5 (orthorhombic, space group P2(1)) is far from O(+) and close to S(+). The solid phase structure of 3 and 5 show small deviation from planarity for the four aromatic rings, whereas two of the aromatic rings in 4 are out of heteroaromatic ring. Spectrophotometric studies in methanol solution revealed that the ligand 3 can be successfully applied to selective amino acid recognition. PMID- 11925211 TI - Rearrangement of 2-aryl-3,3-dichloroazetidines: intermediacy of 2-azetines. AB - An easy synthesis of 2-aryl-3,3-dichloroazetidines, a rather unexplored class of azaheterocycles, is described. The title compounds were easily obtained by reduction of the corresponding 4-aryl-3,3-dichloro-2-azetidinones with monochloroalane, which in turn were synthesized by a ketene-imine [2 + 2] cycloaddition. The reactivity of 3,3-dichloroazetidines with bases was investigated, yielding 2-[dimethoxy(aryl)methyl]aziridines by ring contraction when treated with sodium methoxide. Furthermore, reacting the 3,3 dichloroazetidines with sodium hydride in DMSO, followed by aqueous workup, afforded 1-alkyl-2-aroylaziridines, by hydrolysis of the intermediate 2-azetines and ring closure of the transient 3-amino-2-chloro-1-phenyl-1-propanone derivatives. Monitoring this reaction in an NMR tube, using sodium hydride in DMSO-d(6), allowed the characterization of the intermediate strained heterocyclic enamines, i.e., 2-azetines, by (1)H and (13)C NMR. PMID- 11925212 TI - Ipso substitution as a route to benzo[c]quinolizines and 4-hydroxycoumarins. AB - A convenient ipso substitution method for the preparation of benzo[c]quinolizine (2) and 4-hydroxy-3-(2'-pyridyl)coumarin (3) has been developed. The intramolecular nucleophilic substitution reaction of 3-oxo-2-(2'-pyridyl)-(2 halophenyl)propanoate (1) in refluxing xylenes gives initially benzo[c]quinolizine, while further heating results in the formation of 4 hydroxycoumarin. A mechanism has been proposed to rationalize the two competitive reaction pathways, and the role of HCl is discussed. Under optimized conditions, seven benzo[c]quinolizines and five coumarins were prepared in moderate to good yields. PMID- 11925213 TI - Investigation of the binding of epimer A of the covalent hydrate of 6,7 bis(trifluoromethyl)-8-D-ribityllumazine to a recombinant F22W Bacillus subtilis lumazine synthase mutant by (15)N[(19)F] REDOR NMR. AB - The two epimeric covalent hydrates A and B of 6,7-bis(trifluoromethyl)-8-D ribityllumazine are metabolically stable analogues of hypothetical intermediates proposed in the reactions catalyzed by riboflavin synthase and lumazine synthase. To confirm the stereochemical assignments previously based solely on results for epimer B, a (15)N[(19)F] REDOR NMR study was performed on the complex formed from epimer A and a recombinant, uniformly (15)N-labeled F22W mutant of Bacillus subtilis lumazine synthase. The results indicate that the fluorines of the ligands are closer to the side chain nitrogens of Arg127 and farther away from the side chain nitrogens of Lys135 in epimer B than in epimer A. These results are consistent with the assignment of the earlier 7R configuration of epimer A and the 7S configuration of epimer B. PMID- 11925215 TI - Asymmetric N1 unit transfer to olefins with a chiral nitridomanganese complex: novel stereoselective pathways to aziridines or oxazolines. AB - Chiral nitridomanganese complex 1 was found to be a highly potential N1 unit source for the asymmetric synthesis of aziridines and 2-oxazolines from olefins such as styrene and its derivatives. When sulfonyl chlorides were employed as activators of the complex in the presence of pyridine, pyridine N-oxide, and a silver salt, the reaction of olefins with complex 1 proceeded smoothly to afford the N-sulfonylated aziridines. The aziridination of styrene derivatives with complex 1 using 2-trimethylsilylethanesulfonyl chloride (SESCl) gave the N-SES aziridines, which were easily converted into chiral N-unsubstituted aziridines. It was found that the reaction was applicable to the asymmetric synthesis of 2 oxazolines from olefins when acyl chlorides were employed as activators. Complex 1 provided an effective asymmetric environment for trans-disubstituted styrenes in the reaction (up to 92% ee). This is the first example of a direct asymmetric synthesis of 2-oxazolines from olefins. Additional experiments, conducted during the course of this investigation, suggest that the isomerization of the N acylaziridine intermediate is involved in this reaction. PMID- 11925214 TI - Studies on novel peptidomimetics having bi-directional dispositions of hydroxylated D-Pro-Gly motifs anchored on a C(2)-symmetric iminosugar-based foundation. AB - A rigid pyrrolidine based scaffold comprising of 2,5-dideoxy-2,5-imino-D-idaric acid (1) is developed. Attachment of peptide strands to the carboxylic groups at both ends of this novel template led to the peptidomimetics 2 and 3. Conformational analysis by NMR studies revealed that compounds 2b, 3b and 2c, 3c take interesting turn structures (C(2) symmetric for 2c and 3c) in DMSO-d(6) consisting of identical intramolecular hydrogen bonds at two ends between LeuNH - > sugar-OH as depicted in structure A, whereas 2a and 3a display structures with regular beta-turns with hydrogen bonds between LeuNH --> Boc-C=O in one-half of their molecular frameworks (structure B), characteristic of the turn structures commonly observed in "D-Pro-Gly"-containing peptides. These results suggest that a cis hydroxyl group at the 3-position of the proline residue favors a pseudo beta-turn-like nine-membered ring structure in hydroxyproline-containing peptides involving an intramolecular hydrogen bond between the hydroxyl and the i + 2 backbone amide. PMID- 11925217 TI - Hydroxy-directed diastereoselective installation of a methyl group on indalone models and spiroketal potential precursors for the bafilomycin A(1) C15-C25 subunit. AB - Current efforts devoted to the synthesis of Bafilomycin A(1) led us to investigate a synthetic route through a spiroketal intermediate for the construction of the C15-C25 subunit. Preliminary studies for the diastereoselective installation of the methyl-16 cis with respect to the vicinal OH-15 group through radical opening of either siloxafuran intermediate 7 or cyclopropyl compounds 9 and 13 have been carried out using model compounds derived from commercial Indalone 6. In each case the expected "cis" diastereoisomer was obtained in good to excellent yield. Application of these results to Bafilomycin A(1) synthon led to the opposite "trans" stereoselectivity when alpha-carboxy- or alpha-keto-substituted spiroketals 4 or 19 were used. However, the expected potential intermediate has been obtained from the alpha hydroxymethyl cyclopropanated synthon 21. A Barton-Motherwell xanthate radical deoxygenation-cylopropane opening methodology, followed by a hydroboration oxidation of the exovinylic intermediate, delivered the expected product 22cis in high yield and excellent stereoselectivity. PMID- 11925218 TI - 3,3-Disubstituted allyl alcohols from palladium-catalyzed coupling of hydroaluminated propargyl alcohols with aryl iodides. AB - 3,3-Disubstituted prop-2-en-1-ols 4 were synthesized with a high degree of stereoselectivity from 3-substituted prop-2-yn-1-ols 1 via hydroalumination and subsequent Pd-catalyzed coupling with aryl iodides 3. PMID- 11925216 TI - Glaser-mediated synthesis and photophysical characterization of diphenylbutadiyne linked porphyrin dyads. AB - The Pd-mediated Glaser coupling of a zinc monoethynyl porphyrin and a magnesium monoethynyl porphyrin affords a mixture of three 4,4'-diphenylbutadiyne-linked dyads comprised of two zinc porphyrins (Zn-pbp-Zn), two magnesium porphyrins (Mg pbp-Mg), and one metalloporphyrin of each type (Zn-pbp-Mg). The latter is easily isolated due to the greater polarity of the magnesium versus the zinc chelate. Exposure of Zn-pbp-Mg to silica gel results in selective demetalation, affording Zn-pbp-Fb where Fb = free base porphyrin. This synthesis route employs the magnesium porphyrin as a latent form of the Fb porphyrin, thereby avoiding copper insertion during the Glaser reaction, and as a polar entity facilitating separation. The absorption spectrum of Zn-pbp-Mg or Zn-pbp-Fb is the sum of the spectra of the component parts, while in each case the fluorescence spectrum upon illumination of the Zn porphyrin is dominated by emission from the Mg or Fb porphyrin. Time-resolved absorption spectroscopy shows that the energy-transfer rate constants are (11 ps)(-1) and (37 ps)(-1) for Zn-pbp-Mg and Zn-pbp-Fb, respectively, corresponding to energy-transfer quantum yields of 0.995 and 0.983, respectively. The calculated Forster through-space rates are (1900 ps)(-1) and (1100 ps)(-1) for Zn-pbp-Mg and Zn-pbp-Fb, respectively. Accordingly, the through bond process dominates for both dyads with a through-bond:through-space energy transfer ratio of > or =97:1. Collectively, the studies show that the 4,4' diphenylbutadiynyl linker supports fast and efficient energy transfer between Zn and Mg or Fb porphyrins. PMID- 11925219 TI - Cycloaddition reaction of 2-azadienes derived from beta-amino acids with electron rich and electron-deficient alkenes and carbonyl compounds. Synthesis of pyridine and 1,3-oxazine derivatives. AB - Functionalized keto-enamines 6 were obtained by nucleophilic addition of enol ethers to the imine moiety of 2-azadienes derived from dehydroaspartic esters 4. Reactions of 2-azadiene 4c containing three electron-withdrawing substituents (CO(2)R) with enol ethers 5 in the presence of lithium perchlorate led to the formation of tetrahydropyridine derivatives 7 in a regio- and stereoselective fashion. 2H-[1,3]-oxazines 10 and pyridine derivatives 12 and 13 were obtained by heterocycloaddition reactions of electron-poor azadienes 4d-g containing two electron-withdrawing substituents (4-O(2)N-C(6)H(4), CO(2)R) in positions 1 and 4 with carbonyl derivatives (ethyl glyoxalate 9a and diethyl ketomalonate 9b) and the electron-deficient olefin tetracyanoethylene 11. PMID- 11925220 TI - Allylic phosphates and allylic phosphinates as electrophiles in efficient silylcupration reactions of acetylenes. AB - Atom-efficient stoichiometric silylcupration reactions of acetylenes followed by electrophilic trapping of the intermediate vinylcopper species with allylic phosphates have been developed. The reaction sequence was also carried out with the use of a catalytic amount of CuCN employing of both allylic phosphates and allylic phosphinates as electrophiles. The methods developed provide an easy access to silylated 1,4-diene systems. PMID- 11925222 TI - Synthesis of new transglycosidically tethered 5'-nucleotides constrained to a highly biologically relevant profile. AB - A new motif for restricting 5'-nucleotides to highly biologically relevant conformations has been developed. The 5',6-oxomethylene transglycosidically tethered versions of uridine 5'-monophosphate and 2'-deoxyuridine 5' monophosphate (1 and 2, respectively) were synthesized in 10-11 steps from their respective natural nucleoside precursors along routes general to the preparation of tethered versions of a wide variety of 5'-nucleotide-based compounds. In both routes, a shelf-stable 6-hydroxymethyl pyrimidine nucleoside 5'-carboxaldehyde is the key intermediate. It exists in a carbohydrate-like fashion in a cyclic hemiacetal form under aprotic conditions. The phosphorylated cyclic hemiacetals 1 and 2 were isolated as binary mixtures of 5'-diastereomers differing principally in the trajectory of the phosphate group with respect to the carbohydrate. By (1)H NMR, both 1 and 2 were demonstrated to be stable to hydrolysis at ambient temperature in D(2)O solution for at least 2 months. The oxomethylene transglycosidic tether as deployed in 1 and 2 leaves all of the native 5' nucleotide molecular recognition sites intact while it restricts the framework to a low-energy anti glycosyl conformation and an extended phosphate disposition. This provides a spatial presence that approximates nearly three-quarters of the protein-bound 5'-nucleotide ligands described in the Protein Data Bank. The tether has a low structural and electronic impact, occupies a region of space (over the beta-face of the furan ring) seldom penetrated by proteins, and should be accommodated as readily on purine-based 5'-nucleotide frameworks as on pyrimidine-based ones. Because of its unique and attractive features, this new motif for the conformational restriction of 5'-nucleotides is expected to be useful for producing probes of structure/function relationships and in assessing the conformational binding requirements that enzymes and receptor sites have for their natural 5'-nucleotide-based ligands. PMID- 11925221 TI - 5-[4-(1-Hydroxyethyl)phenyl]-10,15,20-triphenylporphyrin as a probe of the transition-state conformation in hydrolase-catalyzed enantioselective transesterifications. AB - 5-[4-(1-Hydroxyethyl)phenyl]-10,15,20-triphenylporphyrin (1a) and zinc porphyrin 1b were designed and synthesized to experimentally examine the validity of the transition-state model previously proposed for the lipase-catalyzed kinetic resolution of secondary alcohols. The lipases from Pseudomonas cepacia (lipase PS), Candida antarctica (CHIRAZYME L-2), Rhizomucor miehei (CHIRAZYME L-9), and Pseudomonas aeruginosa (lipase LIP) exhibited excellent enantioselectivity (E >100 at 30 degrees C). Subtilisin Carlsberg from Bacillus licheniformis (ChiroCLEC-BL) also showed high enantioselectivity for 1a (E = 140 at 30 degrees C), and the thermodynamic parameters were determined: DeltaDeltaH = -6.8 +/- 0.8 kcal mol(-1), DeltaDeltaS = -13 +/- 3 cal mol(-1) K(-1). Lipases and subtilisin showed R- and S-preference for 1, respectively. The mechanisms underlying the experimental observations are explained in terms of the transition-state models. The large secondary alcohol 1 is a powerful tool for investigating the conformation of the transition state of the enzyme-catalyzed reactions. The fact that 1 was resolved with high enantioselectivity strongly suggests that the gauche conformation, but not the anti conformation, is taken in the transition state, in agreement with the transition-state models involving the stereoelectronic effect. PMID- 11925223 TI - Investigation of synthetic hosts that model cation-pi sites found at protein binding domains. AB - Small cyclophanes containing aromatic groups and dialkyl ammonium ions were created as model systems of the cation-pi complexes found at some protein binding domains. The hosts had different shapes in order to investigate the effect the arrangement of ammonium ions to aromatic surfaces has on their reactivity. pK(a) values of the hosts were substantially different in DMSO or (95/5) DMSO/D(2)O solutions, which showed that the ions existed in different environments of the hosts. Electrostatic charges, as determined by density functional calculations, revealed that the magnitude of a cationic charge depends on its position relative to an aromatic ring. Association constants of the hosts bound to the sodium salt of N-acetyl phenylalanine in d(6)-DMSO and in (95/5) d(6)-DMSO/D(2)O solutions were inversely proportional to the magnitude of the hosts' acidity constants. These results suggest that the magnitude of the positive charge for cationic groups of cation-pi complexes is reduced by being associated with electron-rich faces of aromatic rings. The aromatic rings, however, lessen the desolvation penalty that must be overcome for ligand binding, giving an overall more favorable association. PMID- 11925224 TI - Structure/activity study of tris(2-aminoethyl)amine-derived translocases for phosphatidylcholine. AB - Sulfonamide and amide derivatives of tris(aminoethyl)amine (TREN) are known to facilitate phospholipid translocation across vesicle and erythrocyte membranes; that is, they act as synthetic translocases. In this report, a number of new TREN based translocases are evaluated for their abilities to bind phosphatidylcholine and translocate a fluorescent phosphatidylcholine probe. Association constants were determined from (1)H NMR titration experiments, and translocation half-lives were determined via 7-nitrobenz-2-oxa-1,3-diazol-4-yl (NBD)/dithionite quenching assays. A rough correlation exists between translocase/phosphatidylcholine association constants and translocation half-lives. The tris-sulfonamide translocases are superior to the tris-amide versions because they associate more strongly with the phospholipid headgroup. The stronger association is due to the increased acidity of the sulfonamide NHs as well as a molecular geometry (as shown by X-ray crystallography) that is able to form tridentate complexes with one of the phosphate oxygens. Two fluorescent translocase analogues were synthesized and used to characterize membrane partitioning properties. The results indicate that the facilitated translocation of phospholipids by TREN derived translocases is due to the formation of hydrogen-bonded complexes with the phospholipid headgroups. In the case of zwitterionic phosphatidylcholine, it is the neutral form of the translocases that rapidly associates with the phosphate portion of the phosphocholine headgroup. Complexation masks the headgroup polarity and promotes diffusion of the phospholipid-translocase complex across the lipophilic interior of the membrane. PMID- 11925225 TI - A highly enantioselective hetero-Diels-Alder reaction of aldehydes with Danishefsky's diene catalyzed by chiral titanium(IV) 5,5',6,6',7,7',8,8' octahydro-1,1'-bi-2-naphthol complexes. AB - The catalytic effect of chiral Lewis acids on the hetero-Diels-Alder reaction between aldehydes and Danishefsky's diene (1) has been investigated. A variety of combinations of different ligands and Lewis acids have been examined as catalysts for the hetero-Diels-Alder reaction between benzaldehyde and 1, and it has been found that the readily accessible Ti(IV)-H(8)-BINOL (TiHBOL) complex is a very effective catalyst for the reaction, leading to products with very high enantioselectivity (up to 99% ee) and yield (92%). The hetero-Diels-Alder reaction of other aldehydes with 1 under the catalysis of TiHBOL is a general reaction which proceeds well with very high enantioselectivity and isolated yield for various aldehydes at 0 degrees C to room temperature. Based on the experimental results, the proposed mechanism of the hetero-Diels-Alder reaction and the dihedral angle effects of ligands are discussed. PMID- 11925226 TI - Addition of bromine chloride and iodine monochloride to carbonyl-conjugated, acetylenic ketones: synthesis and mechanisms. AB - The reactions of 3-butyn-2-one (1), 3-hexyn-2-one (2), and 4-phenyl-3-butyn-2-one (3) with bromine chloride (BrCl) and iodine monochloride (ICl) in CH(2)Cl(2), CH(2)Cl(2)/pyridine, and MeOH are described. The data show that the major products in CH(2)Cl(2) are (Z)-AM (anti-Markovnikov) regioisomers. With the exception of 3 and ICl, the (E)-AM regioisomers predominate when pyridine was added as an acid scavenger. Minor amounts of the M regioisomers were formed with 1 and 2 and BrCl. The percentage of M regioisomer increased significantly with 1 and BrCl in MeOH, but MeOH had little affect on the other reactions. Isolation and stability of the products are discussed. Detailed evidence for the structures of the products, involving a combination of MS, (1)H and (13)C NMR, and IR, is presented; HRMS analyses are provided as proofs for all of the products. The acid catalyzed mechanism and the halonium ion mechanism are considered as possible pathways in the formation of the products. PMID- 11925227 TI - Nucleophilic catalysis with 1,8-diazabicyclo[5.4.0]undec-7-ene (DBU) for the esterification of carboxylic acids with dimethyl carbonate. AB - 1,8-Diazabicyclo[5.4.0]undec-7-ene (DBU) is an effective nucleophilic catalyst for carboxylic acid esterification with dimethyl carbonate (DMC). The reaction pathway of this new class of nucleophilic catalysis has been studied. A plausible, multistep mechanism is proposed, which involves an initial N-acylation of DBU with DMC to form a carbamate intermediate. Subsequent O-alkylation of the carboxylate with this intermediate generates the corresponding methyl ester in excellent yield. In the absence of DBU or in the presence of other bases, such as ammonium hydroxide or N-methylmorpholine, the same reaction affords no desired product. This method is particularly valuable for the synthesis of methyl esters that contain acid-sensitive functionality. PMID- 11925228 TI - Synthesis of 1,2,7,7a-tetrahydro-1aH-cyclopropa[b]quinoline-1a-carboxylic acid derivatives, doubly constrained ACC derivatives, by a remarkable cyclopropanation process. AB - Reaction of N-benzoyl-1,2,3,4-tetrahydroquinoline-2-carboxylic acid with acetic anhydride resulted in 1H,3H,5H-oxazolo[3,4-a]quinolin-3-one derivative 13. Different cyclopropanation processes were applied to 13, but only diazomethane in the presence of water furnished the hitherto unknown methyl 1,2,7,7a-tetrahydro 1aH-cyclopropa[b]quinoline-1a-carboxylate 14, which can be considered as a doubly constrained 1-aminocyclopropane-1-carboxylic acid system. The mechanism of the cyclopropanation was studied in detail. The new ACC ester 14 was transformed into fused tetracyclic hydantoin derivatives, which comprised a new type of heterocyclic system. PMID- 11925229 TI - Synthesis of some unsymmetrical bridged terpyridines. AB - Novel unsymmetrical terpyridines 1 and 2 are synthesized using intra- and intermolecular Michael additions as the key reactions, followed by the construction of the central pyridine ring. Terpyridine 1 represents a heretofore unknown hexacyclic ring system. PMID- 11925230 TI - Novel chiral biferrocene ligands for palladium-catalyzed allylic substitution reactions. AB - Eleven novel aminophosphine ligands have been synthesized, all of which contain a chiral 2,2' '-bridged biferroceno unit as part of a biferrocenoazepine substructure. The efficiency of these compounds as chiral auxiliaries in palladium-mediated allylic substitution reactions has been investigated. Depending on the degree of (steric) fit between proper ligands and cyclic or noncyclic substrates, reactions with 46-87% ee were achieved. The molecular structure of a palladium dichloride complex of one of the ligands was determined by X-ray diffraction and compared to its binaphthyl analogue. In the solid state, the azepine substructure of these two complexes adopts totally different conformations with either local C(2) (binaphthyl) or local C(1) (biferrocene derivative) symmetry. These structural changes are well-reproduced by empirical force field calculations and are also reflected in significantly different behavior in asymmetric catalysis. PMID- 11925231 TI - Transition-state variation in the nucleophilic substitution reactions of aryl bis(4-methoxyphenyl) phosphates with pyridines in acetonitrile. AB - The kinetics and mechanism of the reactions of Z-aryl bis(4-methoxyphenyl) phosphates, (4-MeOC(6)H(4)O)(2)P(=O)OC(6)H(4)Z, with pyridines (XC(5)H(4)N) are investigated in acetonitrile at 55.0 degrees C. In the case of more basic phenolate leaving groups (Z = 4-Cl, 3-CN), the magnitudes of beta(X) (beta(nuc)) and beta(Z) (beta(lg)) indicate that mechanism changes from a concerted process (beta(X) = 0.22-0.36, beta(Z) = -0.42 to -0.56) for the weakly basic pyridines (X = 3-Cl, 4-CN) to a stepwise process with rate-limiting formation of a trigonal bipyramidal pentacoordinate (TBP-5C) intermediate (beta(X) = 0.09-0.14, beta(Z) = -0.08 to -0.28) for the more basic pyridines (X = 4-NH(2), 3-CH(3)). This proposal is supported by a large negative cross-interaction constant (rho(XZ) = 1.98) for the former and a positive rho(XZ) (+0.97) for the latter processes. In the case of less basic phenolate leaving groups (Z = 3-CN, 4-NO(2)), the unusually small magnitude of beta(Z) values is indicative of a direct backside attack TBP-5C TS in which the two apical sites are occupied by the nucleophile and leaving group, ap(NX)-ap(LZ). The instability of the putative TBP-5C intermediate leading to a concerted displacement is considered to result from relatively strong proximate charge transfer interactions between the pi-lone pairs on the directly bonded equatorial oxygen atoms and the apical bond (n(O)(eq) - sigma(ap)). These are supported by the results of natural bond orbital (NBO) analyses at the NBO-HF/6-311+G//B3LYP/6-311+G level of theory. PMID- 11925232 TI - Synthesis and photochromism of spirobenzopyrans and spirobenzothiapyran derivatives bearing monoazathiacrown ethers and noncyclic analogues in the presence of metal ions. AB - Spirobenzopyrans bearing monoazathiacrown ethers and noncyclic analogues were synthesized, and their ion-responsive photochromism depending on the dual metal ion interaction with the crown ether and the phenolate anion moieties was examined using alkali and alkaline-earth metal ions, Ag(+), Tl(+), Pb(2+), Hg(2+), and Zn(2+). The prepared spirobenzopyrans showed a selective binding ability to Mg(2+) and Ag(+) with negative and positive photochromism, respectively. Among the metal ions, only Ag(+) facilitated photoisomerization to the corresponding merocyanine form. Depending on the ring size of the monoazathiacrown ether moieties, soft metal ions such as Hg(2+) and Ag(+) showed significant shifts in the UV-vis absorption spectra, while hard metal ions such as Mg(2+), Zn(2+), and Pb(2+) did not afford any meaningful shift. This result reflects that the monoazathiacrown ether and phenolate anion moieties prefer soft and hard metal ions, respectively. Therefore, the Mg(2+) and Ag(+) selectivities are mainly derived from the phenolate anion and monoazathiacrown ether moieties, respectively. On the other hand, a spirobenzothiapyran bearing 3,9-dithia-6 monoazaundecane showed a remarkable selectivity to Ag(+). PMID- 11925234 TI - Photochemistry of arylidene-beta-ionones: a highly efficient route to novel tricyclic ketones through intramolecular, exoselective photochemical (4 + 2) cycloadditions, occurring only in an aqueous-organic solvent. AB - (E,E)-Arylidene-beta-ionones (1a-f) are converted to 1,7,7-trimethyl-3-(E-2' arylethenyl)-2-oxabicyclo[4.4.0]deca-3,5-dienes (3a-f, approximately 90%) by irradiating in anhydrous solvents. Irradiation of (3a-f) in aqueous methanol results in Z,E-arylidene-beta-ionones (2), through retro-electrocyclization, which undergoes an intramolecular, exo-selective [4 + 2] photocycloaddition leading to 11-(exo)-aryl-1,7,7-trimethyl-tricyclo[4.4.0.1(2,4)]undec-5-ene-3-ones (8a-f, 60-80%). The latter rearrange over silica gel to afford, quantitatively, 5 aryl-7,11,11-trimethyl-tricyclo[5.4.0.0(3,6)]undec-1-ene-4-ones (5a-f). Irradiation of 1a-f in aqueous methanol leads to 8a-f, except in case of 1c,f wherein formation, respectively, of tricyclic ketones 9c (55%) and 9f (80%), derived from photodeconjugation in 2, followed by intramolecular [4 + 2] cycloaddition, is observed. PMID- 11925233 TI - Stereospecificity of an enzymatic monoene 1,4-dehydrogenation reaction: conversion of (Z)-11-tetradecenoic acid into (E,E)-10,12-tetradecadienoic acid. AB - In this article, we report the first stereochemical study of an enzymatic 1,4 dehydrogenation reaction, namely, the transformation of (Z)-11-tetradecenoic acid into (E,E)-10,12-tetradecadienoic acid, involved in the sex pheromone biosynthesis of the moth Spodoptera littoralis. The investigation was carried out using the labeled substrates (R)-[10-(2)H]- and (S)-[10-(2)H]-tridecanoic acids ((R)-2 and (S)-2, respectively) and (R)-[2,2,3,3,13-(2)H(5)]- and (S)-[2,2,3,3,13 (2)H(5)]-tetradecanoic acids ((R)-1 and (S)-1, respectively). Probes (R)-2 and (S)-2 were prepared as described in a previous article.(1) The synthesis of the pentadeuterated chiral substrates (R)-1 and (S)-1 was accomplished by kinetic resolution of the racemic 12-tridecyn-2-ol (6) with immobilized porcine pancreatic lipase. The enantiomerically pure alcohols (R)-6 and (S)-6 were transformed into the final acids (S)-1 and (R)-1, respectively, by a sequence of well-established reactions. The analyses of methanolyzed lipidic extracts from glands incubated separatedly with each individual probe showed that in the transformation of (Z)-11-tetradecenoic acid into (E,E)-10,12-tetradecadienoic acid, both pro-(R) hydrogen atoms at C-10 and C-13 are removed from the substrate. This is the first example reported of a desaturase with pro-(R)/pro (R) stereospecificities that gives rise to (E)-double bonds. A mechanistic explanation for the stereochemical outcome of this reaction is advanced. PMID- 11925235 TI - Experimental and theoretical insights regarding the cycloaddition reaction of carbohydrate-based 1,2-diaza-1,3-butadienes and acrylonitrile. A model case for the behavior of chiral azoalkenes and unsymmetric olefins. AB - A series of carbohydrate-based tetrahydropyridazines are prepared by the hetero Diels-Alder reaction of the chiral 1,2-diaza-1,3-butadienes 1 and 2 with acrylonitrile. Reactions are regiospecific, and the observed diastereoselection is consistent with a preferred attack to the Re face of the heterodiene unit, as the chiral sugar placed at C4 does largely protect the opposite Si face. The stereochemistry of the major cycloadduct 4 has been firmly established by an X ray crystallographic study that, in addition, reveals a conformation placing the cyano group in axial orientation. Cycloadducts such as 9 and 11, in which the axial cyano group and the carbohydrate moiety exhibit a cis relationship, undergo a facile E2 elimination that relieves the steric congestion. A detailed computational study is reported to provide better insight into the factors that influence this asymmetric cycloaddition. A DFT study (B3LYP/6-31G) on a reduced model does correctly predict the regiochemistry observed experimentally, while the facial diastereoselection is modeled at a semiempirical (PM3) level on the parent reagents, thereby accounting for the steric factor provided by the chiral substituent. The calculations also indicate that the axial orientation of the cyano group can be rationalized in terms of a stabilizing anomeric effect. PMID- 11925236 TI - Aromatization of 1,6,7,7a-tetrahydro-2H-indol-2-ones by a novel process. Preparation of key-intermediate methyl 1-benzyl-5-methoxy-1H-indole-3-acetate and the syntheses of serotonin, melatonin, and bufotenin. AB - Imine 7 of 1,4-cyclohexanedione mono-ethylene ketal 6 was reacted with maleic anhydride, affording the cyclized adduct 8. Methyl esterification of 8, accompanied by transacetalization, led to the dihydrooxindole derivative 10. Aromatization of 10 was then accomplished with POCl(3), leading directly to the key-intermediate title compound 11 in 74% yield from ketone 6. Serotonin, melatonin, and bufotenin were then obtained by standard reactions. PMID- 11925237 TI - Biosynthetic precursors of the lipase inhibitor lipstatin. AB - Three putative intermediates in the biosynthesis of the lipase inhibitor lipstatin were synthesized in stable isotope-labeled form and were added to fermentation cultures of Streptomyces toxytricini. Biosynthetic lipstatin was isolated and analyzed by NMR spectroscopy. [3,10,11,12-(2)H]-(3S,5Z,8Z)-3 hydroxytetradeca-5,8-dienoic acid (9) was shown to serve as a direct biosynthetic precursor of lipstatin. [7,8-(2)H(2)]Hexylmalonate (11) was also incorporated into lipstatin, albeit at a relatively low rate. The leucine moiety of [(13)C formyl,(15)N]-N-formylleucine (10) was diverted to lipstatin under loss of the (13)C-labeled formyl residue. PMID- 11925238 TI - Total synthesis of gibbilimbols A-D. AB - Gibbilimbols A-D (1-4) were synthesized in 32-49% yield over four steps from commercially available starting materials. A copper-catalyzed coupling of 4 methoxyphenylmagnesium bromide with various unsaturated alkyl bromides was the key step in assembling the (long-chain alkyl)phenol skeleton. PMID- 11925239 TI - Spectral properties and absolute rate constants for beta-scission of ring substituted cumyloxyl radicals. A laser flash photolysis study. AB - A laser flash photolysis study of the spectral properties and beta-scission reactions of a series of ring-substituted cumyloxyl radicals has been carried out. All cumyloxyl radicals display a broad absorption band in the visible region of the spectrum, which decays on the microsecond time scale, leading to a strong increase in absorption in the UV region of the spectrum, which is attributed to the corresponding acetophenone formed after beta-scission of the cumyloxyl radicals. The position of the visible absorption band is red-shifted by the presence of electron-donating ring substituents, while a blue-shift is observed in the presence of electron-withdrawing ring substituents, suggesting that + R ring substituents promote charge separation in the excited cumyloxyl radical through stabilization of the partial positive charge on the aromatic ring of an incipient radical zwitterion. Along this line, an excellent Hammett-type correlation between the experimentally measured energies at the visible absorption maxima of the cumyloxyl radicals and sigma(+) substituent constants is obtained. A red-shift is also observed on going from MeCN to MeCN/H(2)O for all cumyloxyl radicals, pointing toward a specific effect of water. The ring substitution does not influence to a significant extent the rate constants for beta-scission of the cumyloxyl radicals, which varies between 7.1 x 10(5) and 1.1 x 10(6) s(-1), a result that suggests that cumyloxyl radical beta-scission is not governed by the stability of the resulting acetophenone. Finally, k(beta) increases on going from MeCN to the more polar MeCN/H(2)O 1:1 for all cumyloxyl radicals, an observation that reflects the increased stabilization of the transition state for beta-scission through increased solvation of the incipient acetophenone product. PMID- 11925240 TI - Substituent effects on the thermal cis-to-trans isomerization of 1,3 diphenyltriazenes in aqueous solution. AB - The thermal cis-to-trans isomerization of some symmetrically p,p'-disubstituted 1,3-diphenyltriazenes has been studied by means of laser-flash photolysis techniques. The geometric isomerization is catalyzed by general acids and general bases as a result of acid/base-promoted 1,3-prototropic rearrangements. Acid catalysis becomes more prominent as the electron-donating character of the para substituent increases, while base catalysis becomes more important as the electron-withdrawing character of the para substituent increases. In addition, the rate ascribed to the interconversion of neutral cis rotamers through hindered rotation around the nitrogen-nitrogen single bond is found to decrease as the electron-withdrawing character of the para substituent increases. Rates of interconversion of neutral cis rotamers are also found to decrease with decreasing solvent polarity, which is indicative of the involvement of a polar transition state. On the other hand, kinetic investigations of the acid-catalyzed decomposition of target triazenes are consistent with an A1 mechanism. PMID- 11925241 TI - Daphnicyclidins J and K, unique polycyclic alkaloids from Daphniphyllum humile. AB - Two Daphniphyllum alkaloids with unprecedented polycyclic skeletons, daphnicyclidins J (1) and K (2), have been isolated from the stems of Daphniphyllum humile, and the structures and relative stereochemistry were elucidated on the basis of spectroscopic data. The absolute stereochemistry of 1 was established by chemical correlation with a known-related alkaloid, daphnicyclidin D (3), through a modified Polonovski reaction. PMID- 11925242 TI - Cephalocyclidin A, a novel pentacyclic alkaloid from Cephalotaxus harringtonia var. nana. AB - A novel alkaloid with an unprecedented fused-pentacyclic skeleton and six consecutive asymmetric centers, cephalocyclidin A (1), has been isolated from the fruits of Cephalotaxus harringtonia var. nana, and the structure was elucidated on the basis of spectroscopic data. The relative and absolute stereochemistry of 1 was determined by a combination of NOESY correlations, X-ray crystallographic data, and the exciton chirality method. PMID- 11925243 TI - Tuning of regioselectivity in the coupling reaction involving allenic/propargylic palladium species. AB - Two different types of coupling patterns for the Pd(0)-catalyzed coupling reaction of allenic/propargylic zinc reagents with organic halides or propargylic carbonates (acetate) with the corresponding organometallic reagents were observed. After studying the controlling factors on the regioselectivity of this reaction, we demonstrated that the steric hindrance of both reactants and the types of organic halides determine the regioselectivity of this coupling reaction. By subtle choosing of the substrates, the regioselectivity can be tuned. On the basis of these results, new methodologies for the highly regio- and stereoselective synthesis of 6-substituted hex-5-yn-2-enoates and 4,6-dialkylhexa 2,4,5-trienoates have been developed. Some of the products synthesized by the carbonate protocol cannot be prepared by the lithiation protocol because the regioselectivity of lithiation of dialkyl-substituted internal alkynes is an intrinsic problem. PMID- 11925244 TI - Ab initio study of 1,4-pentadienyl electrocyclic reactions. AB - The thermochemistry and transition states of the electrocyclic ring closures of the resonance-stabilized 1,4-pentadienyl radical to cyclopenten-3-yl, cyclobut-2 enylmethyl, and 2-vinylcyclopropyl are investigated at Hartree-Fock and coupled cluster levels of theory. The CCSD(T)//QCISD/cc-pVDZ calculations predict activation barriers of 130, 169, and 236 kJ/mol, respectively, and DeltaH values of -60, 115, and 155 kJ/mol. Experimental evidence for the appearance of vinylcyclopropyl following photolytic generation of pentadienyl is more likely the result of a distinct electrocyclic reaction than quenching of a two-step mechanism for formation of cyclopentenyl. Higher energy pathways for formation of polycyclic structures are also briefly examined. PMID- 11925245 TI - Kinetics of hydrolysis of 8-(arylamino)-2'-deoxyguanosines. AB - The 8-(arylamino)-2'-deoxyguanosines, or C-8 adducts, are the major adducts formed by reaction of N-arylnitrenium ions derived from carcinogenic and mutagenic amines with 2'-deoxyguanosine (d-G) and guanosine residues of DNA. The hydrolysis kinetics of three C-8 adducts 1a-c were determined by UV and HPLC methods at 20 degrees C under acidic, neutral, and mildly alkaline conditions. At pH < 2 the dominant hydrolysis process is spontaneous cleavage of the C-N bond of the doubly protonated substrate, 1H(2)(+2) (Scheme 2). The C-8 adducts are 2- to 5-fold more reactive than d-G under these conditions. At 3 < pH < 6 the hydrolysis kinetics are dominated by cleavage of the C-N bond of the monoprotonated nucleoside 1H(+). Under these conditions the hydrolysis kinetics are accelerated by 40- to 1300-fold over that of d-G. The rate increase appears to be caused by a combination of steric acceleration of C-N bond cleavage and a decrease in the ionization constant of 1H(+), K(a1), due to the electron-donating properties of the arylamino C-8 substituent. Under neutral pH conditions a slow (k(obs) approximately 10(-8) s(-1) to 5 x 10(-7) s(-1)) spontaneous cleavage of the C-N bond of the neutral nucleoside, 1, occurs that has not been previously reported for simple purine nucleosides. Finally, under mildly alkaline conditions a process consistent with spontaneous decomposition of the anion 1(-) or OH(-) induced decomposition of 1 is observed. The latter process has been observed for other purine nucleosides, including the closely related 1d, and involves nucleophilic attack of OH(-) on C-8 to cleave the imidazole ring of the purine. PMID- 11925246 TI - Chiral tricyclic iminolactone derived from (1R)-(+)-camphor as a glycine equivalent for the asymmetric synthesis of alpha-amino acids. AB - The development of a highly efficient and stereoselective methodology for the preparation of alpha-amino acids is described. The chiral template, tricyclic iminolactone 7, was synthesized from (1R)-(+)-camphor in five steps in 50% overall yield. Alkylation of iminolactone 7 afforded the alpha-monosubstituted products in good yields (74-96%) and excellent diastereoselectivities (>98%). Hydrolysis of the alkylated iminolactones furnished the desired alpha-amino acids in good yields and enantioselectivities with nearly quantitative recovery of the chiral auxiliary 4. PMID- 11925247 TI - Novel [2 + 2] photocycloaddition-induced rearrangement of bichromophoric naphthalene-tethered resorcinol ethers. AB - The first examples of sequential photocycloaddition-rearrangement reactions of naphthalene-tethered resorcinol ethers are described. Bichromophoric aromatic compounds with naphthalene and resorcinol ether moieties were irradiated in the presence/absence of a small amount of acid to give the corresponding cycloaddition-rearrangement products. From the determination of quantum yields, steady-state fluorescence spectral studies, and fluorescence lifetime measurements, the mechanism of this novel photoinduced multistep reaction was elucidated to involve the initial intramolecular exciplex formation, followed by the intramolecular [2 + 2] photocycloaddition between the two aromatic rings and the subsequent acid-catalyzed skeletal rearrangement of the resulting cyclobutane derivative leading to the final products. PMID- 11925248 TI - Decarbonylative radical cyclization of alpha-amino selenoesters upon electrophilic alkenes. A general method for the 6-azabicyclo[3.2.1]octane synthesis. AB - alpha-Amino selenoester-tethered electronically poor alkenes on treatment with tributyltin hydride or TTMSS undergo intramolecular radical cyclization to provide 6-azabicyclo[3.2.1]octanes through 1-aminomethyl radical intermediates. PMID- 11925249 TI - Titanium isopropoxide as efficient catalyst for the aza-Baylis-Hillman reaction. Selective formation of alpha-methylene-beta-amino acid derivatives. AB - The direct formation of alpha-methylene-beta-amino acid derivatives is achieved using the aza version of the Baylis-Hillman protocol. The products are readily formed in a three-component one-pot reaction between arylaldehydes, sulfonamides, and alpha,beta-unsaturated carbonyl compounds. The reaction is efficiently catalyzed by titanium isopropoxide and 2-hydroxyquinuclidine in the presence of molecular sieves. The protocol allows for structural variation of the substrates, tolerating electron-poor and electron-rich arylaldehydes and various Michael acceptors. PMID- 11925250 TI - Highly diastereoselective aziridination of imines with trimethylsilyldiazomethane. Subsequent silyl substitution with electrophiles, ring opening, and metalation of C-silylaziridines--a cornucopia of highly selective transformations. AB - Treatment of a range of N-sulfonyl (Ts and SES) imines derived from aromatic, heteroaromatic, aliphatic, and unsaturated aldehydes with trimethylsilydiazomethane gave C-silylaziridines in good yield (32-83%) and with high diastereoselectivity in favor of the cis product (80:20-100:0). In contrast, an alpha-imino ester gave predominantly the trans-aziridine (89:11) in high yield (91%). The synthetic potential of C-silylaziridines was investigated. Treatment with F(-) (tetrabutylammonium triphenyldifluorosilicate was used) in the presence of aldehydes gave the alpha-hydroxyaziridines in high yield and high diastereoselectivity (86:14-98:2) for the newly created stereogenic center. Complete retention of configuration was observed in the substitution of the silyl group with electrophiles in all cases. Trapping with deuterium (using CDCl(3) as electrophile) was also successful, but trapping with phosphate [using ClP(O)(OPh)(2)] and acetate (using Ac(2)O) was unsuccessful. In these latter cases ring opening by chloride and acetate, respectively, was observed. Further ring-opening reactions were effected using azide and thiolate nucleophiles and in all cases complete regioselectivity in favor of attack at the silyl-bearing carbon occurred. Complete regioselectivity was also observed in the carbonylative ring expansion using Co(2)(CO)(8) to give a beta-lactam. Treatment of cis-1-tosyl 2-phenyl/butyl-3-trimethylsilylaziridines with n-BuLi and subsequent quenching with MeI followed completely different pathways, depending on the 2-substituent. In the case of the 2-phenylaziridine, metalation was initiated alpha to the phenyl group and led finally to a fused tricyclic adduct with four stereogenic centers as a single diastereoisomer. In the case of the 2-butylaziridine, metalation occurred alpha to the silyl group and led to a trisubstituted silylaziridine, probably via an azirine intermediate. PMID- 11925251 TI - A general method for the preparation of 4- and 6-azaindoles. AB - Nitropyridines reacted with an excess of vinyl Grignard reagent to produce 4- or 6-azaindoles. Improved yields were obtained when a halogen atom was present at the position alpha to the nitrogen atom in the pyridine ring. PMID- 11925252 TI - Pyrene-armed calix[4]azacrowns as new fluorescent ionophores: "molecular taekowndo" process via fluorescence change. AB - A series of new calixarene-based fluoroionophores were synthesized. With our new calixarene derivative bearing a crown ether and an azacrown ether as two binding sites, the metal ion was found to selectively choose its better binding pocket between these two ligands. Interesting "molecular taekowndo" processes between Ag(+)-K(+), Cu(2+)-K(+), and Ag(+)-Cs(+) pairs were easily monitored via fluorescence change. PMID- 11925253 TI - Synthesis of syn and anti isomers of trans-cyclopropyl arginine. AB - There is currently considerable interest in arginine and its structural analogues in the context of nitric oxide synthase (NOS) substrates and inhibitors. Of particular interest are conformationally constrained arginine analogues used to probe the active sites of the three NOS isoforms. A simple procedure is described for the preparation of syn- and anti-trans-cyclopropyl arginine starting from the alpha-OBO-protected Cbz-dehydroglutamate. Cyclopropanation is effected by diazomethane addition followed by irradiation of the resulting pyrazoline and gives a 3:1 mixture of syn:anti isomers that can be separated by crystallization. Reduction of the ester to the alcohol followed by guanylation gives the fully protected cyclopropyl arginine analogues. The CBZ protecting groups are removed by hydrogenolysis and the OBO by mild acid treatment followed by base hydrolysis. PMID- 11925255 TI - Direct synthesis of 5-substituted naphthoquinones. AB - Peri-metalation of 4-(dimethylamino)-1-tert-butyldimethylsilyloxynaphthalene (5) followed by reaction with an electrophile and Jones' oxidation affords 5 substituted naphthoquinones. PMID- 11925254 TI - First total synthesis of artepillin C established by o,o'-diprenylation of p halophenols in water. AB - We have demonstrated that prenylation of p-halophenols was dependent on the solvent effect and succeeded in o,o'-diprenylation of p-halophenols in water. Following the Mizoroki-Heck coupling of the diprenyl-p-iodophenol 3c with methyl acrylate and then hydrolysis, we first synthesized artepillin C [3-(4-hydroxy-3,5 di(3-methyl-2-butenyl)phenyl)-2(E)-propenoic acid] (1), which is a biologically active constituent of propolis. These reactions may be applicable to the synthesis of various useful natural products such as 2,4,6-trisubstituted phenol derivatives. PMID- 11925256 TI - A convenient preparation of functionalized 1,8-dioxygenated naphthalenes from 6 alkoxybenzocyclobutenones. AB - An alternate route for the synthesis of naphthalene building blocks 1-4 has been developed, starting from readily available 6-alkoxybenzocyclobutenones. As thermolysis of a propynylbenzocyclobutenol only provided the naphthalene in low yield, allenyl-substituted benzocyclobutenols were investigated. The desired allenic precursors were prepared by a two-step procedure, which involved hydroxyl directed reduction of the chloropropargylbenzocyclobutenols obtained from addition of lithiopropargyl chloride to the benzocyclobutenones. Thermolysis of the allenic alcohols gave the desired naphthalenes in good yields. PMID- 11925258 TI - Preparation of 2,6-dimethyl-4-arylpyridine- 3,5-dicarbonitrile: a paired electrosynthesis. AB - Electrolysis of benzylthiocyanate, benzyl chloride, p-methylbenzyl chloride, p methoxybenzyl chloride, or toluene in acetonitrile, at platinum electrodes in a two compartments cell divided by a glass-frit diaphragm, affords 2,6-dimethyl-4 arylpyridine-3,5-dicarbonitrile as major product. PMID- 11925257 TI - Palladium-catalyzed carbonylative annulation of o-alkynylphenols: syntheses of 2 substituted-3-aroyl-benzo[b]furans. AB - We report here a general synthetic methodology for palladium-catalyzed carbonylative annulation of o-alkynylphenol to construct 2-substituted-3-aroyl benzo[b]furan. On the basis of the results, this methodology could be applied to a wider selection of iodide substrates to generate desired products. In accordance with mechanistic studies, this process involves coordination of cationic and less hindered acyl palladium complexes with o-alkynylphenols to create a desired cascade triad (coordination, nucleophilic addition, and reductive elimination). Consistent with this mechanism, addition of 1 equiv of AgBF(4) to palladium catalyst Pd(Ph(3)P)(4) generates an ideal candidate for this unique transformation. PMID- 11925259 TI - Unique charge-separated pyridinium-barbituric acid zwitterions. AB - A synthetic procedure for the preparation of the unusual charge-separated pyridinium barbiturate zwitterion 2 from 1,3-dimethylbarbituric acid and 2 pyridinecarbaldehyde in methanol was developed. The structure of the compound was confirmed with X-ray analysis to demonstrate the strong charge separation throughout the molecule. One would expect that this charge separation would increase its reactivity; however, contrary to this expectation, the compound is very stable in acidic media, and in the presence of a base, decarbonylation occurs on one barbituric acid while the zwitterionic moiety of the molecule stays intact. PMID- 11925260 TI - Regioselective lithiation and functionalization of 3-(benzyloxy)isothiazole: application to the synthesis of thioibotenic acid. AB - Direct functionalization of the 3-oxygenated isothiazole heteroaromatic parental system has not yet been reported in the literature. Here, we report the first regioselective lithiation of the 5-position of 3-(benzyloxy)isothiazole (4) using LDA in diethyl ether. The versatility of the methodology was explored by quenching with a variety of electrophiles to give the desired products 7a,b,d-g in 54-68% yield. Only benzoylation aiming at the synthesis of 7c was unsuccessful. Furthermore, a highly convergent synthesis of thioibotenic acid (1), the sulfur analogue of the neurotoxic natural product ibotenic acid, was carried out. PMID- 11925261 TI - Novel acid-catalyzed rearrangement of tetrahydro-1,2,3,4-tetrazines: unexpected formation of glycosazones. AB - The present contribution discloses a simple and unexpected acid-catalyzed cleavage of tetrahydrotetrazines leading to 1,2-bis(hydrazones). Incorporation of a chiral fragment derived from carbohydrates enables the rapid preparation of glycosazones, a family of compounds employed by Emil Fischer to elucidate the configuration of sugars. In addition, a mechanistic proposal accounts for experimental observations. PMID- 11925262 TI - Imidazopyridinium and pyridopyrimidium bromides: synthesis and hydrolysis. AB - The reactions of symmetrical and unsymmetrical 2,2'-dipyridylamines with 1,2 dibromoethane and 1,3-dibromopropane give imidazopyridinium and pyridopyrimidium bromides, respectively. These acetone/CH(2)Cl(2)-insoluble, highly fluorescent quaternary ammonium salts undergo addition/ring opening upon treatment with methanolic KOH to give pyridin-2-one derivatives. A sequential N,N dialkylation/ring-opening hydrolysis/N,N-dialkylation/ring-opening hydrolysis strategy was developed for the construction of unsymmetrical bis(pyridin-2-ones). PMID- 11925264 TI - Recent reviews. 64. PMID- 11925265 TI - Social support for families of children with mental retardation: comparison between Korea and the United States. AB - The informal and professional supports available for families of children with mental retardation in Korea and the United States were examined and the effects of culture and social support on maternal stress considered. Thirty-eight American and 40 Korean mothers participated in the home-visit interview. A pilot study identified 10 types of informal and 11 types of professional support for the two national groups. Results showed that American mothers received more informal and professional support in almost all domains of social support; Korean mothers experienced more stress. Possible explanations for these differences were explored. PMID- 11925266 TI - Assessing secondary conditions among adults with developmental disabilities: a preliminary study. AB - Pope (1992) asserted that there was a significant need to examine secondary conditions among individuals with development disabilities. In the present study we focused on that need. The development of a secondary conditions surveillance instrument is described, as are the results of a pilot survey conducted with adults receiving state developmental disabilities program supports and with their direct-care service providers. Results of a pilot survey are presented to illustrate how survey data might be used to improve systems of services and supports to enhance the health and participation of adults with developmental disabilities in community life. PMID- 11925267 TI - Employment and income status of adults with developmental disabilities living in the community. AB - A comprehensive national portrait of employment and income status of adults with developmental disabilities was estimated through secondary analyses of the 1990 and the 1991 Survey of Income and Program Participation. Results indicate that the majority of adults with developmental disabilities had very limited economic resources, even when earnings from employment and benefits from governmental income support programs were both included. The minority, who worked in a variety of occupations, were earning higher incomes than previous estimates. The overwhelming majority were unemployed. Implications of the low-income profile and potential underutilization of employment services was discussed. PMID- 11925268 TI - Medicaid HCBS Waivers and supported employment pre- and post-Balanced Budget Act of 1997. AB - Findings from a national survey of state mental retardation/developmental disability agencies regarding use of the Medicaid Home and Community Based Waiver to fund supported employment were reported. Numbers of individuals and funding levels were requested for day habilitation services for FYs 1997 and 1999, before and after the(P.L. 105-33), which removed eligibility restrictions for this service. Findings show that growth rates for this service far exceeded growth rates for other day services, with high growth rates in a small number of states. However, supported employment accounted for less than 16% of those receiving day habilitation services through the Waiver and only 12% of day habilitation funding, with the remainder going to day support, prevocational services, and other segregated options. PMID- 11925269 TI - Coping strategies and the impact of challenging behaviors on special educators' burnout. AB - Although challenging behaviors have been identified as a source of staff stress, few researchers have directly addressed this relationship. In the present study, 55 teachers and support staff in special schools for children with mental retardation completed questionnaires assessing burnout, coping strategies for challenging behavior, and their exposure to challenging behavior. Results showed that (a) use of maladaptive coping strategies for challenging behaviors constitutes a risk for staff burnout, (b) this risk is in addition to that associated with exposure to challenging behavior, and (c) use of maladaptive coping strategies moderated the impact of exposure to challenging behaviors on emotional exhaustion burnout. Implications for future research and for the support of staff working with individuals who have challenging behaviors are discussed. PMID- 11925270 TI - Factor stability of the Schalock and Keith (1993) Quality of Life Questionnaire. AB - In the 1990s, there was a significant change in how governments viewed publicly provided services. In the area of disability services, it has been suggested that providers could demonstrate their effectiveness with reference to the quality of life of their clients. One instrument often used in quality of life research for people with intellectual disabilities is the Quality of Life Questionnaire; however, before this instrument can be used with confidence, the reliability of its scores must be demonstrated. We investigated the stability of the four Quality of Life Questionnaire factors over various populations. Three of the four factors were found to be stable. This raises potential concern over the use of the Quality of Life Questionnaire in assessing service providers' effectiveness. PMID- 11925271 TI - Advancing research in decision-making capacity: an opportunity for leadership-and an obligation of geriatric psychiatry. PMID- 11925272 TI - Involving decisionally impaired subjects in research: the need for legislation. PMID- 11925273 TI - Provisional diagnostic criteria for depression of Alzheimer disease. AB - The authors, a group of investigators with extensive research and clinical experience related to both late-life depression and Alzheimer disease (AD), propose provisional affective and behavioral inclusion and exclusion diagnostic criteria for Depression of AD. PMID- 11925274 TI - Provisional diagnostic criteria for depression of Alzheimer disease: rationale and background. AB - This review provides the rationale and background for the development of diagnostic criteria for depression of Alzheimer disease (AD), including risk factors and neurobiological correlates, epidemiology, and clinical characteristics, along with course, assessment, treatment, economics, a description of the criteria, and future research directions. Overall, there is substantial research to suggest that the depression that may co-occur with AD is different from other depressive disorders. Further research is needed to better define core symptoms, clinical course, and efficacy of treatments. PMID- 11925275 TI - Improving understanding of research consent in middle-aged and elderly patients with psychotic disorders. AB - Individuals with schizophrenia may show impaired capacity to make decisions about participating in research, yet these patients also show considerable heterogeneity in decisional abilities. Problems with procedures contribute to patients' difficulties in understanding consent forms. Few studies have focused on improving comprehension of research consent in older patients with psychotic disorders. In this study, 80 middle-aged and elderly outpatients with schizophrenia or related psychotic disorders and 19 normal comparison subjects were randomized to receive a routine consent (RC) or enhanced consent (EC) procedure. The EC procedure consisted of a computerized slide show incorporating more structure and review of important information. A comprehension test was administered after the consent procedure; subjects were given up to three trials of the post-test to answer all of the questions correctly. Overall, the normal comparison subjects obtained a higher score on the post-consent comprehension test than the patients. Within each of these two groups, those who received EC had better comprehension than those who received RC. Interestingly, EC patients did not differ significantly from RC normal comparison subjects in their post test scores. Among the patients, comprehension test scores correlated with level of education and cognitive performance. PMID- 11925276 TI - Current state of research on decision-making competence of cognitively impaired elderly persons. AB - As the number of cognitively impaired elderly persons increases, the need for evidence-based assessments of their capacity to consent to medical treatment and research participation will grow. The authors conducted an electronic and manual literature search for all English-language articles examining the decision-making capacity of elderly persons with dementia or cognitive impairment, reviewing articles in relation to key areas of methodological, clinical, and policy importance. The 32 relevant studies identified were highly heterogeneous, even in their definitions and measurements of decisional capacity. Although incapacity is common, many persons with dementia are capable of making their own medical and research decisions. In Alzheimer disease, memory and executive-function deficits predict decisional impairment. Still, at least in early stages of dementia, interventions may improve decisional abilities. Short and simple cognitive screening may be useful by identifying persons in need of more intensive evaluations. The use of expert judgment-based methods may mitigate the problem of a lack of a criterion standard for competence. Research into the decision-making competence of cognitively impaired elderly persons is a growing field. It is beginning to yield findings with practical implications for preserving the autonomy and welfare of this group of vulnerable elderly patients. PMID- 11925277 TI - The California Scale of Appreciation: a new instrument to measure the appreciation component of capacity to consent to research. AB - Capacity to consent is one of the linchpins of the ethical conduct of clinical care and research, and it needs to be reliably measured. The authors describe the development of a new measure of the "appreciation" component of capacity, the California Scale of Appreciation (CSA), 18 items rated according to the concept of "patently false belief" (a belief that is grossly improbable); 39 patients with schizophrenia or a related psychotic disorder (27 outpatients and 12 inpatients) and 15 normal-comparison subjects participated. Each subject's audiotaped interview was rated by three evaluators. Answers to each item were scored as "capable," "incapable," or "uncertain capacity." Also, each subject was given an overall rating of one of these three categories by each rater. Total scores on the CSA were calculated and correlated with scores on standardized instruments for assessing psychopathology and cognitive impairment. The mean total CSA score was significantly lower in the patients than in the normal comparison subjects; however, a majority of the patients were found to be fully "capable" on the CSA. The CSA is a potentially useful instrument for measuring the appreciation component of capacity in persons with psychotic disorders. Its generalizability to other patient populations and to other types of protocols needs to be determined. PMID- 11925278 TI - Similarities and differences in depression among black and white nursing home residents. AB - The authors examined racial similarities and differences in depressive symptomatology, diagnosis, and the predictors of depression in four independent nursing homes, conducting analyses across all sites and separately for the nursing home with the greatest racial balance (NH4). All-site data indicated that white residents showed more depression than black residents. There were no racial differences in the depression diagnosis derived from a structured interview of DSM-III-R. At NH4, there were no statistically significant racial differences in any of the measures of depression. Across sites, functional disability was the strongest predictor of both Geriatric Depression Scale (GDS) and DSM-III-R diagnosis of depression in both blacks and whites. Cognitive impairment and use of antidepressants were predictive of medical chart diagnosis of depression across sites, but not of depression measured by GDS or DSM-III-R criteria. At NH4, functional disability was predictive of GDS depression, but only among whites. Age was not an important predictor of depression. Results indicate the importance of considering the method used to diagnose depression and the necessity of controlling for the nursing home setting when examining racial differences in depression. PMID- 11925280 TI - New postoperative depressive symptoms and long-term cardiac outcomes after coronary artery bypass surgery. AB - The authors evaluated the impact of an increase in depressive symptoms at 6 months after elective coronary artery bypass graft surgery on long-term cardiac morbidity and mortality between 6 and 36 months postoperatively. Patients who had low scores for depressive symptomatology pre-operatively and who completed follow up at 6 months were contacted again 36 months after surgery to assess cardiac and neurologic morbidity and mortality. At 36 months after surgery, an interval history was completed, and baseline questionnaires were readministered. Follow-up was obtained on 123/124 patients (99%). The rate of combined new cardiac morbidity/mortality between 6 and 36 months was 13.6% among those with newly increased depressive symptoms at 6 months vs. 3.0% in the patients without new depressive symptoms at 6 months. Only an increase in depressive symptoms at 6 months was related to the occurrence of subsequent cardiac complications between 6 and 36 months. In this small sample of patients, increased depressive symptoms at 6 months after surgery appear to be associated with the occurrence of subsequent major cardiac morbidity/ mortality. PMID- 11925279 TI - Allelic differences in the serotonin transporter-linked polymorphic region in geriatric depression. AB - Previous studies have examined the role of genetic variations in the serotonin transporter-linked polymorphic region (5HTTLPR) in affective disorders. The authors studied 182 older depressed subjects and 107 elderly control subjects and obtained DNA for genotyping at the 5HTTLPR. There were no significant differences in allele frequencies generally or for number of short alleles for the group as a whole, but interesting gender effects emerged. Among men, 23% of depressed men had two short alleles, compared with only 5% of control subjects. Among women, 67% of depressed women with more than one episode had at least one short allele, compared with 41% of single-episode female patients. Also, 74% of women with a positive family history of psychiatric illness in any female relative had at least one short allele, whereas 53% had at least one short allele who did not have such a family history. Our results add to the literature linking this gene to affective illness. The negative association of allele frequency and depression may be related to the relatively small sample size. The findings raise the possibility that this genetic locus may exert differential effects based on gender, increasing risk in men, and increasing risk of recurrence in women. PMID- 11925281 TI - Effects of nortriptyline and paroxetine on postural sway in depressed elderly patients. AB - Tricyclic antidepressants (TCAs) have previously been found to be related to an increased incidence of falls in elderly persons. Recent pharmacoepidemiologic and nursing home studies have suggested that the risk of falls and fractures in elderly patients receiving selective serotonin reuptake inhibitors (SSRIs) is not different from that of patients receiving TCAs. The authors therefore evaluated postural sway in an older population of depressed patients randomly assigned to treatment with either nortriptyline or paroxetine and did not find any change in postural sway after 6 weeks' treatment with either antidepressant. Further studies with other SSRIs are needed. PMID- 11925282 TI - Treatment-related decision-making capacity in middle-aged and older patients with psychosis: a preliminary study using the MacCAT-T and HCAT. AB - The authors conducted a preliminary examination of treatment-related decision making capacity (DMC) in middle-aged and older psychosis patients. The MacArthur Competency Assessment Tool for Treatment (MacCAT-T) was administered to measure DMC among 16 patients with psychotic disorders and without dementia (mean age: 54.6 [SD 7.2] years). The impact of repeated learning trials on patients' understanding was assessed with the Hopkins Competency Assessment Test (HCAT). Subjects were also assessed with the Positive And Negative Syndrome Scale and Mattis' Dementia Rating Scale. There was a wide range of performance on the MacCAT-T; the levels and range of MacCAT-T scores were similar to those previously reported among younger acutely hospitalized patients in the MacCAT-T validation study. Patients' understanding of disclosed material on the HCAT improved significantly over repeated presentations. Although there were no statistically significant correlations observed between DMC and demographic characteristics, severity of symptoms, or global cognitive deficits, the sample size provided limited power to detect such associations. Results point to the importance of assessing DMC and to the potential modifiability of initial difficulties in understanding. PMID- 11925283 TI - Ondansetron in the treatment of cognitive decline in Alzheimer dementia. AB - The authors performed a multicenter, double-blind, placebo-controlled clinical trial to assess the efficacy and safety of ondansetron (20 microg/day and 100 microg/ day) in treating cognitive decline in 185 patients with Alzheimer disease. Although ondansetron was well tolerated without any serious drug-related side effects, the study failed to demonstrate any significant cognitive improvement. PMID- 11925284 TI - A mother as caregiver of a patient with Alzheimer disease. PMID- 11925285 TI - Hierarchy and heritability: the role of diagnosis and modeling in psychiatric genetics. PMID- 11925286 TI - The illness of Vincent van Gogh. AB - Vincent van Gogh (1853-1890) had an eccentric personality and unstable moods, suffered from recurrent psychotic episodes during the last 2 years of his extraordinary life, and committed suicide at the age of 37. Despite limited evidence, well over 150 physicians have ventured a perplexing variety of diagnoses of his illness. Henri Gastaut, in a study of the artist's life and medical history published in 1956, identified van Gogh's major illness during the last 2 years of his life as temporal lobe epilepsy precipitated by the use of absinthe in the presence of an early limbic lesion. In essence, Gastaut confirmed the diagnosis originally made by the French physicians who had treated van Gogh. However, van Gogh had earlier suffered two distinct episodes of reactive depression, and there are clearly bipolar aspects to his history. Both episodes of depression were followed by sustained periods of increasingly high energy and enthusiasm, first as an evangelist and then as an artist. The highlights of van Gogh's life and letters are reviewed and discussed in an effort toward better understanding of the complexity of his illness. PMID- 11925287 TI - The human genome: chromosome 22q11 deletion syndrome. PMID- 11925288 TI - Psychological versus biological clinical interpretation: a patient with prion disease. PMID- 11925290 TI - A twin study of genetic relationships between psychotic symptoms. AB - OBJECTIVE: Biometrical model fitting was applied to clinical data from twins to investigate whether operationally defined schizophrenic, schizoaffective, and manic syndromes share genetic risk factors. METHOD: Seventy-seven monozygotic and 89 same-sex dizygotic twin pairs in which the proband met the Research Diagnostic Criteria (RDC) for lifetime-ever schizophrenic, schizoaffective, or manic syndrome were ascertained from the Maudsley Twin Register in London. The syndromes were defined non-hierarchically. Correlations in liability were calculated for each syndrome in the monozygotic and dizygotic pairs and across the three pairings of schizophrenic-manic, schizophrenic-schizoaffective, and schizoaffective-manic syndromes both within probands and within pairs. For the three syndromes considered together, an independent pathway model was fitted. RESULTS: The model fitting showed significant genetic correlations between all three syndromes. There was evidence of both common and syndrome-specific genetic contributions to the variance in liability to the schizophrenic and manic syndromes, but the genetic liability to the schizoaffective syndrome was entirely shared in common with the other two syndromes. In contrast, environmental liability to the schizoaffective syndrome was not shared with the other syndromes. CONCLUSIONS: If diagnostic hierarchies are relaxed, there is a degree of overlap in the genes contributing to RDC schizophrenic, schizoaffective, and manic syndromes. Supplementing the traditional approach of assigning a single main lifetime diagnosis with information on within-person comorbidity of psychotic syndromes may provide valuable information about the familial aggregation of psychotic symptoms. PMID- 11925291 TI - Impaired cortical network for preattentive detection of change in speech sounds in schizophrenia: a high-resolution event-related potential study. AB - OBJECTIVE: Schizophrenia is associated with language-related dysfunction. At the most basic level, the dysfunction of schizophrenic patients in perceiving phoneme boundaries has been demonstrated in a behavioral task. The goal in this study was to obtain an electrophysiological index of preattentive detection of change in speech sounds by schizophrenic patients. METHOD: A high-density recording system was used to measure mismatch negativity, i.e., difference in electrophysiological responses between standard and deviant auditory stimuli. The amplitude of mismatch negativity was measured in 23 right-handed schizophrenic patients and in 28 healthy comparison subjects. Three types of mismatch negativity were recorded: change in duration of pure-tone stimuli, change in duration of the Japanese vowel /a/, and difference between vowels /a/ and /o/ (across-phoneme change). RESULTS: The schizophrenic patients had a lower amplitude of mismatch negativity for across-phoneme change than for change in duration of tone or vowel. For across phoneme change, the schizophrenia group showed significantly lower bilateral amplitude of mismatch negativity than the comparison subjects. These findings coincide with those from the analysis of scalp current density, indicating that the schizophrenic patients had a significantly weaker left temporal combination of current sink (perpendicularly sinking into the scalp) and current source (arising out of the scalp) and a significantly weaker right frontal/temporal current sink than the comparison subjects. CONCLUSIONS: These results demonstrate impaired frontotemporal cortical networks for preattentive detection of change in speech sounds in schizophrenia. The language-related dysfunction in schizophrenia may be present at the early stage of auditory processing of relatively simple stimuli, such as phonemes, and not just at stages involving higher-order semantic processes. PMID- 11925292 TI - A multidose study of haloperidol decanoate in the maintenance treatment of schizophrenia. AB - OBJECTIVE: Maintenance medication is critical in the prevention of psychotic relapse and rehospitalization among patients with schizophrenia. Given potential adverse effects, identification of the minimum effective dose is clinically important. METHOD: A multicenter, double-blind study was conducted to determine rates of symptomatic exacerbation and adverse effects in 105 outpatients with schizophrenia randomly assigned to four different fixed doses of haloperidol decanoate and treated for 1 year or until relapse. The doses used were 25, 50, 100, and 200 mg given intramuscularly once per month. RESULTS: Rates of symptomatic exacerbation were 15% in the 200-mg group, 23% with 100 mg, 25% with 50 mg, and 60% with 25 mg. No significant differences in outcome were found between the groups treated with 200, 100, and 50 mg. Among the patients who completed the trial with no symptomatic exacerbation, there were no differences between dose groups on measures of psychopathology at the final rating point. CONCLUSIONS: The results of this study suggest that the 200-mg/month dose of haloperidol decanoate is associated with the lowest rate of symptomatic exacerbation (15%) with minimal increased risk of adverse effects or subjective discomfort in comparison to 100 or 50 mg. At the same time, the rates of worsening with 100 mg (23%) and 50 mg (25%) were not significantly greater than that seen with 200 mg. These results provide some guidance as to effective dose ranges of haloperidol decanoate. PMID- 11925293 TI - Association of diabetes mellitus with use of atypical neuroleptics in the treatment of schizophrenia. AB - OBJECTIVE: The development of both type I and type II diabetes after initiation of some atypical neuroleptics has been reported, primarily in studies involving small series of patients. This study used administrative data from a large national sample of patients with a diagnosis of schizophrenia to compare the prevalence of diabetes mellitus in patients receiving prescriptions for atypical and typical neuroleptics. METHOD: All outpatients with schizophrenia treated with typical and atypical neuroleptics over 4 months in 1999 in the Veterans Health Administration of the Department of Veterans Affairs (VA) were included in this study. Patients treated with atypical neuroleptics were those who received prescriptions for clozapine, olanzapine, risperidone, or quetiapine. Patients with a diagnosis of diabetes were also identified by using ICD-9 codes in VA administrative databases. The prevalence of diabetes mellitus across age groups and among patients receiving prescriptions for different atypical neuroleptics was examined with multiple logistic regression. RESULTS: A total of 38,632 patients were included in the study: 15,984 (41.4%) received typical neuroleptics and 22,648 (58.6%) received any atypical neuroleptic (1,207 [5.3%] received clozapine; 10,970 [48.4%], olanzapine; 955 [4.2%], quetiapine; and 9,903 [43.7%], risperidone; 387 patients received prescriptions for more than one atypical neuroleptic). When the effects of age were controlled, patients who received atypical neuroleptics were 9% more likely to have diabetes than those who received typical neuroleptics, and the prevalence of diabetes was significantly increased for patients who received clozapine, olanzapine, and quetiapine, but not risperidone. However, for patients less than 40 years old, all of the atypical neuroleptics were associated with a significantly increased prevalence of diabetes. CONCLUSIONS: In this large group of patients with schizophrenia, receipt of a prescription for atypical neuroleptics was significantly associated with diabetes mellitus. PMID- 11925294 TI - Racial disparities in antipsychotic prescription patterns for patients with schizophrenia. AB - OBJECTIVE: This study aimed to examine the extent and type of variation in antipsychotic prescription patterns between African American and Caucasian patients with schizophrenia. METHOD: Subjects were 2,515 adult Medicaid recipients treated for schizophrenia in 1995 with one of four types of antipsychotic medication (traditional antipsychotics, clozapine, risperidone, or depot antipsychotics). Prescription and mental health service use data were collected from Medicaid claims files for the 12 months following the first filled antipsychotic prescription. Patterns of antipsychotic prescription were compared for African American (N=1,538, 61%) and Caucasian (N=977, 39%) subjects. RESULTS: African American subjects were significantly younger and more likely to receive Supplemental Security Income than were the Caucasian subjects, who received mental health services more continuously. African American subjects were less likely than Caucasian subjects to receive clozapine (8% versus 15%, respectively) and risperidone (25% versus 31%) and more likely to receive depot antipsychotics (26% versus 14%). The likelihood of receiving clozapine or risperidone remained significantly different after demographic and service use characteristics were controlled. CONCLUSIONS: This study found ethnic disparities in antipsychotic prescription patterns among a large number of publicly insured clients treated for schizophrenia. Given the rapidly changing pharmacological treatment environment, these findings have significant implications for differential quality of care for African American patients. Future studies employing client and provider characteristics are urgently needed to test alternative explanations for ethnic disparities. PMID- 11925295 TI - Patient and psychiatrist ratings of hypothetical schizophrenia research protocols: assessment of harm potential and factors influencing participation decisions. AB - OBJECTIVE: A central ethical issue in schizophrenia research is whether participants are able to provide informed consent, particularly for protocols entailing medication washouts or placebo treatments. Few data show how patients with schizophrenia and psychiatrists assess such scientific designs regarding potential harm, willingness to participate, and the relative influence of clinicians, family members, and financial incentives upon participation decisions. METHOD: In this preliminary study, structured interviews were conducted with schizophrenia patients (N=59), and parallel surveys were completed by attending and resident psychiatrists (N=70). Four hypothetical research protocols were rated. Patients were asked about their own views; psychiatrists provided both their personal views and predictions of patient views. RESULTS: Patients and psychiatrists both perceived substantially different levels of harm across the four protocols, identifying significantly greater harm for medication washouts or placebo treatments. Participants were less willing to enroll in protocols perceived as more harmful. Schizophrenia patients found enrollment decisions relatively easy. Patients and psychiatrists indicated that doctor recommendations, monetary incentives, and, to a lesser extent, family preferences had a mild influence on participation decisions. CONCLUSIONS: Given hypothetical protocols with variable design elements, schizophrenia patients and psychiatrists made meaningful and discerning harm assessments and participation decisions. These findings suggest that schizophrenia patients may have strengths in the research consent process that may not be fully recognized. The impact of outside influences upon research enrollment decisions remains uncertain. While psychiatrists were often accurate in predicting patient responses, data suggest the importance of clarifying views of individual patients regarding specific protocols. PMID- 11925296 TI - Views of potential subjects toward proposed regulations for clinical research with adults unable to consent. AB - OBJECTIVE: The authors' goal was to assess healthy individuals' attitudes toward five of the most prominent proposed safeguards regarding the consent process for research with adults unable to consent. METHOD: Telephone interviews were conducted with 246 individuals with a family history of Alzheimer's disease who had participated in clinical research. RESULTS: The majority of respondents said that they were willing to participate in research if they lost the ability to consent. Few completed a research advance directive. Many had discussed their preferences with their families, and the majority would allow their families to make research decisions for them. CONCLUSIONS: Enrolling individuals who are unable to consent in research that offers no potential for medical benefit is consistent with the preferences of at least some individuals. This suggests that such research should not be prohibited, provided there is sufficient evidence that it is consistent with the preferences of individual subjects. Requiring that such evidence be provided in a formal research advance directive may be unnecessarily restrictive. More research is needed to assess whether the findings in this group of subjects generalize to other groups. PMID- 11925297 TI - Obsessive-compulsive and panic symptoms in patients with first-admission psychosis. AB - OBJECTIVE: The occurrence, persistence and specificity of the association between comorbid obsessive-compulsive and panic symptoms and three psychotic disorders- schizophrenia/schizoaffective disorder, bipolar disorder with psychosis, and major depression with psychosis--were examined in a first-admission, epidemiologically defined group of patients with psychotic symptoms. METHOD: The Structured Clinical Interview for DSM-III-R obsessive-compulsive and panic modules were administered at baseline and 24-month follow-up to patients with schizophrenia/schizoaffective disorder (N=225), bipolar disorder with psychosis (N=138), and major depression with psychosis (N=87) participating in the Suffolk County (N.Y.) Mental Health Project. The rates of subsyndromal symptoms and disorder criteria met were compared across the three psychosis groups. Recognition and treatment of anxiety symptoms at initial discharge and impact of the baseline presence of anxiety symptoms on 24-month clinical status were also examined. RESULTS: Obsessive-compulsive and panic symptoms were present at baseline in 10%-20% of all three groups. There was no specific association between obsessive-compulsive symptoms and any specific psychosis diagnosis; however, women with major depression with psychosis had a significantly higher rate of panic symptoms than the other two groups, and schizophrenia/schizoaffective disorder patients with baseline panic symptoms were significantly more likely to exhibit positive symptoms of psychosis after 24 months. CONCLUSIONS: The authors found no specific association between obsessive compulsive symptoms and diagnosis early in the illness course, but the finding of an association between panic symptoms and psychotic depression among female patients and between baseline panic and positive psychotic symptoms in schizophrenia/schizoaffective disorder patients at 24 months suggests the need for further study. PMID- 11925298 TI - Serotonin transporter distribution and density in the cerebral cortex of alcoholic and nonalcoholic comparison subjects: a whole-hemisphere autoradiography study. AB - OBJECTIVE: Lesions of the medial prefrontal cortex and dysfunctions in serotonin turnover are two well-established factors associated with impulsive and sociopathic behaviors, but no firm neuroanatomical data have linked these pathophysiological findings. The aims of this study were to identify putative areas in the human brain that are rich in serotonin transporter sites, particularly within the medial prefrontal cortex, and to determine whether serotonin transporter density in this area is altered among alcoholic subjects. METHOD: Serotonin transporter density was measured among 17 alcoholic and 10 nonalcoholic comparison subjects by postmortem whole-hemisphere autoradiography with [(3)H]citalopram. RESULTS: In the human cerebral cortex, serotonin transporter binding sites were concentrated in the perigenual anterior cingulate cortex. Substantially sparser serotonin transporter density (up to 35%) was observed in the perigenual anterior cingulate cortex of alcoholic subjects in relation to nonalcoholic comparison subjects. After adjustment for age and postmortem delay, this finding remained statistically significant. CONCLUSIONS: A lower serotonin transporter density among the alcoholic subjects was observed, specifically in the so-called "affect" region, suggesting an association between ethanol addiction and dysfunctional serotonergic neurotransmission in this area. PMID- 11925299 TI - Family functioning and peer affiliation in children of fathers with antisocial personality disorder and substance dependence: associations with problem behaviors. AB - OBJECTIVE: Family functioning and peer influences are theoretically linked to child psychopathology. This study quantified the functional status of families with fathers with substance dependence with or without comorbid antisocial personality disorder and evaluated the peer environments of preadolescent offspring. The authors examined associations between the child's psychopathology, paternal substance dependence/antisocial personality disorder status, and measures of family and peer environments. METHOD: Families with the presence or absence of paternal substance dependence were subdivided into those with and without paternal antisocial personality disorder. Grouped families were contrasted on measures of family functioning, the child's peer affiliation, and the child's problem behaviors. Regression analysis determined the influence of these factors on the child's psychopathology. RESULTS: Families with paternal substance dependence functioned worse than normal comparison families. However, families with paternal substance dependence and antisocial personality disorder (N=34) did not differ markedly from those with substance dependence without antisocial personality disorder (N=84). The children of fathers with both substance dependence and antisocial personality disorder had greater affiliation with deviant peers than those with substance dependence without antisocial personality disorder and comparison families (N=104). CONCLUSIONS: Children of fathers with substance dependence and antisocial personality disorder demonstrated higher externalizing and internalizing psychopathology than those with substance dependence but not antisocial personality disorder and those without either condition. Paternal substance dependence/antisocial personality disorder status and the child's affiliation with deviant peers were most robustly associated with the child's psychopathology. Research is needed to develop interventions that effectively address parental risk and healthy peer relations. PMID- 11925300 TI - Age, gender, and ethnicity differences in patterns of cocaine and ethanol use preceding suicide. AB - OBJECTIVE: The goal of this study was to examine the relationship between suicide and substance use among completed suicides in Fulton County, Ga., from 1994 through 1998. METHOD: Extensive data on completed suicides were obtained from records of the Office of the Medical Examiner. Specific characteristics of the victims who had used cocaine, ethanol, or both before committing suicide were compared across demographic groups. RESULTS: There were important race, sex, and age differences among the victims who had used cocaine and ethanol before committing suicide. Almost all (94.6%) of the suicide victims in whom cocaine was detected were male, 51.4% of the cocaine-positive victims were African American men, and 43.2% were white men. Substance use by teenage victims differed dramatically across ethnic lines. The vast majority (86.7%) of African American teenagers did not use either substance before committing suicide, whereas 50.0% of the white teenage victims had used one or both substances, with 41.7% of the white teenagers having used ethanol. Overall, ethanol use was much more common among white victims of all age groups. CONCLUSIONS: Intoxicant use preceding suicide occurred in specific age-, ethnic-, and gender-based patterns. PMID- 11925301 TI - Risk factors for 12-month comorbidity of mood, anxiety, and substance use disorders: findings from the Netherlands Mental Health Survey and Incidence Study. AB - OBJECTIVE: This study examined risk factor profiles of pure and comorbid 12-month mood, anxiety, and substance use disorder in the general population. METHOD: Data were derived from the Netherlands Mental Health Survey and Incidence Study, a prospective epidemiologic study in which a representative sample of 7,076 adults age 18-64 years were interviewed with the Composite International Diagnostic Interview. Logistic regression was used to compare subjects with a diagnosis of pure and comorbid disorders with non-psychopathological comparison subjects and to compare subjects with comorbid disorders with those with pure disorder on sociodemographic characteristics, chronic somatic conditions, parental psychiatric history, and childhood traumas and adversities. RESULTS: Only 39.5% of the subjects with a 12-month mood disorder, 59.3% of those with an anxiety disorder, and 75.4% of those with a substance use disorder exhibited the disorder in the pure form. Comorbid anxiety and mood disorders, the most prevalent comorbid condition, showed associations with eight of the nine sociodemographic and long-term vulnerability factors investigated; pure mood disorder and pure anxiety disorder were each linked to only about half of the factors. Female gender, younger age, lower educational level, and unemployment were associated with comorbid anxiety and mood disorders but not with pure mood disorders. The risk profiles of pure anxiety disorder and pure substance use disorder similarly diverged from those of the comorbid conditions. CONCLUSIONS: High levels of psychiatric comorbidity exist in the general population. The risk factor profiles for comorbid disorders differ considerably from those for pure disorders. Primary prevention of secondary disorders in populations with a history of a primary disorder are important for reducing psychiatric burden. PMID- 11925302 TI - Frontal white matter biochemical abnormalities in late-life major depression detected with proton magnetic resonance spectroscopy. AB - OBJECTIVE: Neuroanatomical abnormalities have been identified in patients with late-life mood disorders by using magnetic resonance imaging. This study examined the biochemical correlates of late-life major depression in the frontal gray and white matter by using single-voxel proton spectroscopy. METHOD: Twenty elderly patients with major depression and 18 comparison subjects similar in age and gender to the patients were scanned on a 1.5-T magnetic resonance scanner with head coil. Voxels were placed in the left dorsolateral white matter and bilaterally in the anterior cingulate gray matter. Absolute levels of N acetylaspartate, choline, myo-inositol, and creatine were estimated with the LC Model algorithm. Ratios of metabolite to creatine levels were computed from the absolute values. RESULTS: myo-Inositol/creatine and choline/creatine ratios were significantly higher in the frontal white matter in the major depression group than in the comparison group. The groups had no significant differences in the metabolite ratios in the gray matter. CONCLUSIONS: Biochemical changes in the white matter may provide some of the neurobiological substrates to late-life major depression. PMID- 11925303 TI - A descriptive analysis of minor depression. AB - OBJECTIVE: The authors provide a detailed clinical description of minor depression: its symptoms, level of disability, stability, and relationship to patient and family history of major depressive disorder. METHOD: Rigorous criteria for minor depression, including functional disability, were used to identify 226 individuals for a three-phase treatment study. This report presents data obtained on that study group during the first study phase, a 4-week placebo lead-in period. RESULTS: One hundred sixty-two subjects (72% of the initial study group) remained in the study for 4 weeks and continued to meet criteria for minor depression. Minor depression in these subjects was primarily characterized by mood and cognitive symptoms, not the classical neurovegetative signs and symptoms of depression. Approximately one-third of the subjects with minor depression had a past history of major depressive disorder, and nearly half had a family history of unipolar depressive disorder; however, neither factor affected the severity or quality of minor depressive symptoms. CONCLUSIONS: These data suggest that 1) minor depression is not evanescent; 2) minor depression is characterized by mood and cognitive symptoms rather than neurovegetative symptoms; 3) minor depression may occur either independently of a lifetime history of major depressive disorder or as a stage of illness in the course of recurrent unipolar depressive disorder; and 4) depressive disorders should be conceptualized as a continuum of severity. PMID- 11925304 TI - Negative symptoms in temporal lobe epilepsy. AB - OBJECTIVE: This study examined the frequency of negative and positive symptoms in nonpsychotic patients with temporal lobe epilepsy and the relationship of negative and positive symptoms to cognition, quantitative magnetic resonance imaging (MRI) volumetrics, and depression. METHOD: Eighty-four patients with temporal lobe epilepsy and 74 healthy comparison subjects were evaluated for negative and positive symptoms and underwent comprehensive neuropsychological evaluation, quantitative MRI volumetrics, and assessment of mood state and depression. RESULTS: Negative symptoms were significantly more prevalent in the patients with temporal lobe epilepsy (31%) than in the comparison subjects (8%). There was no difference between groups in the rate of positive symptoms. Although the epilepsy patients as a group exhibited generalized cognitive impairment relative to the comparison subjects, the epilepsy patients with negative symptoms performed significantly worse than patients without negative symptoms and comparison subjects across measures of nonverbal intelligence, visuoperception, speeded visuomotor processing, and memory. The epilepsy patients with negative symptoms exhibited significantly greater diffuse atrophy than the healthy comparison subjects and higher CSF volumes than the epilepsy patients without negative symptoms. The epilepsy patients with and without negative symptoms had statistically equivalent Beck Depression Inventory scores and lifetime history of mood disorders, including major depression. CONCLUSIONS: Negative but not positive symptoms were more prevalent in temporal lobe epilepsy patients than in healthy comparison subjects. Negative symptoms were independent of current and past depression and were associated with neuropsychological deficits exceeding the general cognitive morbidity associated with temporal lobe epilepsy and with quantitative MRI indices suggesting greater cerebral atrophy. PMID- 11925305 TI - A functional polymorphism in the COMT gene and performance on a test of prefrontal cognition. AB - OBJECTIVE: In the prefrontal cortex, the enzyme catechol O-methyltransferase (COMT) is critical in the metabolic degradation of dopamine, a neurotransmitter hypothesized to influence human cognitive function. The COMT gene contains a functional polymorphism, Val158Met, that exerts a fourfold effect on enzyme activity. The current study investigated whether prefrontal cognition varies with COMT genotype. METHOD: Val158Met was genotyped in 73 healthy volunteers. A task of prefrontal cognition, the Wisconsin Card Sorting Test, was also administered. RESULTS: Subjects with only the low-activity met allele made significantly fewer perseverative errors on the Wisconsin Card Sorting Test than did subjects with the val allele. CONCLUSIONS: These data are consistent with those of previous studies, suggesting that a functional genetic polymorphism may influence prefrontal cognition. PMID- 11925306 TI - Metformin for weight loss in pediatric patients taking psychotropic drugs. AB - OBJECTIVE: Metformin was assessed as a treatment for weight gain in children taking olanzapine, risperidone, quetiapine, or valproate. METHOD: The subjects were 19 patients aged 10-18 years; 15 were white and four were black, and there were 12 boys and seven girls. In a 12-week open-label study, each patient received metformin, 500 mg t.i.d. Changes in weight and body mass index were evaluated by using repeated measures analysis of variance. RESULTS: Of the 19 patients, 15 lost weight, three gained 1.6 kg or less, and one had no change. The mean changes in weight and body mass index at 12 weeks were highly significant. CONCLUSIONS: Metformin merits further study as a treatment for weight gain in patients taking psychotropic medications. PMID- 11925307 TI - An animal model of Tourette's syndrome. AB - OBJECTIVE: An animal model was used to investigate an autoimmune etiology for some cases of Tourette's syndrome. METHOD: Sera from 12 patients with Tourette's syndrome with high levels of antineural or antinuclear antibodies were infused bilaterally into the ventrolateral striatum of rats. Sera from 12 additional Tourette's syndrome patients and 12 normal subjects (both groups with low levels of autoantibodies) were infused for comparison. Rates of oral stereotypies were recorded by observers who were blind to the origin of the infused sera. RESULTS: Oral stereotypies significantly increased in the rats infused with sera from the patients with high levels of autoantibodies. CONCLUSIONS: The results are consistent with an autoimmune etiology in a subset of cases of Tourette's syndrome. PMID- 11925308 TI - Comparative study of trauma-related phenomena in subjects with pseudoseizures and subjects with epilepsy. AB - OBJECTIVE: The purpose of this study was to examine potential differences in measures of trauma-related phenomena between subjects with pseudoseizures and subjects with intractable epilepsy. METHOD: Thirty-one adult subjects with pseudoseizures and 32 subjects with intractable epilepsy (confirmed by video-EEG) were recruited from the epilepsy unit of a tertiary care hospital. Each participant completed the Impact of Event Scale, the Davidson Trauma Scale, the Mississippi Scale for Combat-Related Posttraumatic Stress Disorder (PTSD), the Dissociative Experience Scale, and the Pittsburgh Sleep Quality Index, as well as demographic, seizure history, and family functioning measures. RESULTS: Subjects with pseudoseizures had significantly higher mean scores on the Davidson Trauma Scale, Mississippi Scale for Combat-Related PTSD, Impact of Event Scale, and Pittsburgh Sleep Quality Index than subjects with epilepsy. In addition, a significantly higher percentage of subjects with pseudoseizures had scores above the clinical cutoff level of 30 on the Dissociative Experience Scale. CONCLUSIONS: Subjects with pseudoseizures exhibited trauma-related profiles that differed significantly from those of epileptic comparison subjects and closely resembled those of individuals with a history of traumatic experiences. Interventions aimed at trauma-related issues may be beneficial for patients with pseudoseizures. PMID- 11925309 TI - Increased occipital cortex GABA concentrations in depressed patients after therapy with selective serotonin reuptake inhibitors. AB - OBJECTIVE: Studies have demonstrated low concentrations of gamma-aminobutyric acid (GABA) in the plasma and CSF of individuals with major depression, and low GABA concentrations have also been found in the occipital cortex of depressed subjects. The goal of this study was to determine whether these occipital cortex GABA concentrations are altered after administration of selective serotonin reuptake inhibitors (SSRIs) in the treatment of depression. METHOD: By means of proton magnetic resonance spectroscopy, occipital cortex GABA concentrations were measured in 11 medication-free depressed patients before initiation of treatment with SSRI medications and after an average of 2 months of treatment. RESULTS: A significant increase in occipital cortex GABA concentrations was seen after SSRI treatment. CONCLUSIONS: SSRI treatment of major depression is associated with increased occipital cortex GABA concentrations. This appears to result in a normalization of low pretreatment GABA concentrations, an effect that may contribute to a common mechanism of antidepressant action. PMID- 11925310 TI - An open trial of morning light therapy for treatment of antepartum depression. AB - OBJECTIVE: About 5% of pregnant women meet criteria for major depression. No pharmacotherapy is specifically approved for antepartum depression; novel treatment approaches may be welcome. The authors explored the use of morning bright light therapy for antepartum depression. METHOD: An open trial of bright light therapy in an A-B-A design was conducted for 3-5 weeks in 16 pregnant patients with major depression. The Hamilton Depression Rating Scale, Seasonal Affective Disorders Version, was administered to assess changes in mood. A follow up questionnaire was used to assess outcome after delivery. RESULTS: After 3 weeks of treatment, mean depression ratings improved by 49%. Benefits were seen through 5 weeks of treatment. There was no evidence of adverse effects of light therapy on pregnancy. CONCLUSIONS: These data provide evidence that morning light therapy has an antidepressant effect during pregnancy. A randomized controlled trial is warranted to test this alternative to medication. PMID- 11925311 TI - Odegaard's selection hypothesis revisited: schizophrenia in Surinamese immigrants to The Netherlands. AB - OBJECTIVE: The incidence of schizophrenia among Surinamese immigrants to the Netherlands is high. The authors tested Odegaard's hypothesis that this phenomenon is explained by selective migration. METHOD: The authors imagined that migration from Surinam to the Netherlands subsumed the entire population of Surinam and not solely individuals at risk for schizophrenia. They compared the risk of a first admission to a Dutch mental hospital for schizophrenia from 1983 to 1992 for Surinamese-born immigrants to the risk for Dutch-born individuals, using the Surinamese-born population in the Netherlands and the population of Surinam combined as the denominator for the immigrants. RESULTS: The age- and sex adjusted relative risk of schizophrenia for the Surinamese-born immigrants was 1.46. CONCLUSIONS: Selective migration cannot solely explain the higher incidence of schizophrenia in Surinamese immigrants to the Netherlands. PMID- 11925312 TI - Serotonin syndrome and atypical antipsychotics. PMID- 11925313 TI - Stress regulation and self-mutilation. PMID- 11925314 TI - Ziprasidone induction of hypomania in depression? PMID- 11925315 TI - Nerve growth factor and smoking cessation. PMID- 11925316 TI - Quetiapine and pregnancy. PMID- 11925317 TI - Creativity and affective illness. PMID- 11925319 TI - Creativity and affective illness. PMID- 11925318 TI - Topiramate for refractory schizophrenia. PMID- 11925321 TI - Buproprion and sexual dysfunction. PMID- 11925323 TI - Creativity and affective illness. PMID- 11925325 TI - Early-stage vision and schizophrenia. PMID- 11925326 TI - Neuropsychiatric testing and the menstrual cycle. PMID- 11925327 TI - Suicide risk in placebo-controlled studies. PMID- 11925329 TI - Panic and depression. PMID- 11925332 TI - Take the time to listen. PMID- 11925348 TI - Cognitive impairment in bipolar affective disorder: implications for the bipolar diathesis. PMID- 11925349 TI - Recruiting and retaining psychiatrists. PMID- 11925350 TI - Famine: the distant shadow over French psychiatry. PMID- 11925351 TI - Protean nature of mass sociogenic illness: from possessed nuns to chemical and biological terrorism fears. AB - BACKGROUND: Episodes of mass sociogenic illness are becoming increasingly recognised as a significant health and social problem that is more common than is presently reported. AIMS: To provide historical continuity with contemporary episodes of mass sociogenic illness in order to gain a broader transcultural and transhistorical understanding of this complex, protean phenomenon. METHOD: Literature survey to identify historical trends. RESULTS: Mass sociogenic illness mirrors prominent social concerns, changing in relation to context and circumstance. Prior to 1900, reports are dominated by episodes of motor symptoms typified by dissociation, histrionics and psychomotor agitation incubated in an environment of preexisting tension. Twentieth-century reports feature anxiety symptoms that are triggered by sudden exposure to an anxiety-generating agent, most commonly an innocuous odour or food poisoning rumours. From the early 1980s to the present there has been an increasing presence of chemical and biological terrorism themes, climaxing in a sudden shift since the 11 September 2001 terrorist attacks in the USA. CONCLUSIONS: A broad understanding of the history of mass sociogenic illness and a knowledge of episode characteristics are useful in the more rapid recognition and treatment of outbreaks. PMID- 11925352 TI - Recruitment into psychiatry. AB - BACKGROUND: Despite improvements in psychiatric teaching, British medical schools have never produced enough graduands aiming for psychiatry. AIMS: To inform the strategy for improving recruitment. METHOD: A literature review. RESULTS: The number of psychiatrists required depends on the role of psychiatry, which is constantly changing. The present requirement is about 250-300 per year, including replacements and new posts. The number of psychiatric trainees has always been higher than expected from the career plans of newly qualified doctors, but the number of British graduates passing the Royal College of Psychiatrists' Membership examination has still fallen short, requiring a supplement of foreign medical graduates. The recent 50% expansion in medical students may make this country self-sufficient. CONCLUSIONS: To improve recruitment, the College should focus on influences before and after undergraduate training - the kind of student entering medical school and the factors favouring sustained psychiatric practice after graduation. PMID- 11925353 TI - Sustained attention deficit in bipolar disorder. AB - BACKGROUND: Recovery in bipolar disorder is central to its definition but is rarely complete. Previous work has suggested that neuropsychological impairment persists during the euthymic state but has been confounded partly by mild affective symptoms in remitted patients. AIMS: To characterise neuropsychological functioning in the euthymic phase of bipolar disorder with an emphasis on tasks of executive functioning. METHOD: Thirty euthymic patients with bipolar disorder were compared with thirty healthy controls on neuropsychological tasks differentially sensitive to damage within prefrontal cortex. RESULTS: Bipolar I patients were impaired on tasks of attentional set shifting, verbal memory and sustained attention. Only sustained attention deficit survived controlling for mild affective symptoms. This deficit was related to progression of illness, but was none the less present in a subgroup of patients near illness onset. CONCLUSIONS: Sustained attention deficit may represent a neuropsychological vulnerability marker for bipolar disorder, providing a focus for further understanding of the phenotype and analysis of the neuronal networks involved. PMID- 11925354 TI - Case-control study of neurocognitive function in euthymic patients with bipolar disorder: an association with mania. AB - BACKGROUND: Neurocognitive impairments in euthymic patients with bipolar disorder may represent trait rather than state variables. AIMS: To test the hypothesis that euthymic patients with bipolar disorder would exhibit impairment in verbal learning and memory and executive function compared with healthy controls matched for age, gender and premorbid IQ. METHOD: Twenty euthymic patients with bipolar disorder were matched, on a case-by-case basis, to twenty healthy community controls. Cases and controls were tested with a battery of neuropsychological tests. RESULTS: Impairments were found in cases compared with controls in tests of verbal learning and memory. Verbal learning and memory correlated negatively with the number of manic episodes. CONCLUSIONS: Impaired verbal learning and memory may be a trait variable in bipolar disease. There are implications for adherence to medication and relapse and for the role of early treatment interventions. Prospective designs and targeting first-episode groups may help to differentiate trait v. disease process effects. PMID- 11925355 TI - Randomised trial of thyroxine to prevent postnatal depression in thyroid-antibody positive women. AB - BACKGROUND: Women who are positive for thyroid antibodies in early gestation are prone to post-partum depression, apparently independent of thyroid dysfunction, as measured by serum levels of free thyroxine, free triodothyroxine and thyroid stimulating hormone. This finding may be due to infrequent monitoring of thyroid function, because hyperthyroidism, hypothyroidism and combinations of both may occur post-partum. AIMS: To test the hypothesis that stabilising thyroid function post-partum by administering daily thyroxine reduces the rate of occurrence and severity of associated depression. METHOD: In a randomised double-blind placebo controlled trial, 100 microg of thyroxine or placebo was given daily to 446 thyroid-antibody-positive women (342 of whom were compliant) from 6 weeks to 6 months post-partum, assessing their psychiatric and thyroid status at 4-weekly intervals. RESULTS: There was no evidence that thyroxine had any effect on the occurrence of depression. The 6-month period prevalence of depression was similar to that reported previously. CONCLUSIONS: The excess of depression in thyroid antibody-positive women in the post-partum period is not corrected by daily administration of thyroxine. PMID- 11925356 TI - Amygdala volume in schizophrenia: post-mortem study and review of magnetic resonance imaging findings. AB - BACKGROUND: Claims that schizophrenia is a disease of the limbic system have been strengthened by meta-analyses of magnetic resonance imaging (MRI) studies finding reduced hippocampus and amygdala volumes. Some post-mortem studies do not find these abnormalities. AIMS: To assess the volume of the amygdala in a series of brains post-mortem. METHOD: Amygdala volume was estimated using point-counting in both hemispheres of the brains of 10 male and 8 female patients with schizophrenia, and a comparison group of 9 males and 9 females. RESULTS: No significant reduction of amygdala volume was found. CONCLUSIONS: Significant volume reduction of the amygdala is not a consistent feature of schizophrenia; findings from early MRI studies using coarse delineation methods may introduce bias to subsequent meta-analyses. PMID- 11925357 TI - Evidence for non-progressive changes in adolescent-onset schizophrenia: follow-up magnetic resonance imaging study. AB - BACKGROUND: It is not clear how far brain abnormalities in early-onset schizophrenia result from progressive neurodevelopmental or neurodegenerative processes. Aims To investigate the hypothesis that structural brain abnormalities in adolescent-onset schizophrenia are progressive in the early phase of the illness. METHOD: A magnetic resonance imaging case-control study of 16 adolescents with schizophrenia (mean age 16.6 years, s.d.=1.9 years) with a mean time of 2.7 years (s.d.=1.7 years) between measurements and 16 matched controls (average age 16.0 years, s.d.=2.0 years) with a mean time of 1.7 years (s.d.=0.5 years) between measurements. RESULTS: There was no evidence of progressive structural brain changes during late adolescence. Significant ventricular enlargement (greater in males) and left-sided temporal lobe changes were evident from the outset of the illness. CONCLUSIONS: Neurodevelopmental brain abnormalities are non-progressive during late adolescence. PMID- 11925358 TI - Enhanced semantic priming in schizophrenia: a computer model based on excessive pruning of local connections in association cortex. AB - BACKGROUND: Many studies have found that people with schizophrenia exhibit abnormally high levels of semantic priming. Post-mortem and neuroimaging studies of schizophrenia suggest a reduction of neuritic processes (dendrites and synapses). AIMS: To demonstrate that reductions in neuritic processes can produce excessive priming in patients with schizophrenia. METHOD: Associative memory was simulated using a computer-based neural network system consisting of two interactive neural groups, one coding for individual memories and the other for the category to which each memory belonged. RESULTS: Variation of a single parameter determining the density of local connections within the two neuronal groups gave a close approximation to levels of memory access and semantic priming previously reported in normal subjects and in patients with schizophrenia. CONCLUSIONS: This study suggests that schizophrenia arises from excessive pruning of local connections in association cortex. Its findings shed light on the mechanisms underlying cognitive priming more generally, and how it might emerge developmentally. PMID- 11925359 TI - Psychosis in high-security and general psychiatric services: report from the UK700 and special hospitals' treatment resistant schizophrenia groups. AB - BACKGROUND: Serious violence is an unusual but significant correlate of psychosis, and leads to the need for specialist secure psychiatric services. Most such service users have previously used general psychiatric services. AIMS: To examine diagnostic and socio-demographic differences between high-security psychiatric service users from their peers in community services. METHOD: Two groups of patients with psychosis were compared: a national sample of high security hospital residents, and a sample of patients in contact with general psychiatric services. RESULTS: Schizophrenia was the almost invariable diagnosis for all special hospital patients. White patients in the community sample were significantly more likely to have affective components to their illness compared with African-Caribbean patients; unlike those in special hospitals. There was a small excess in the proportion of African-Caribbean patients in the special hospital group, controlling for diagnosis, gender and locality. Men were overrepresented in this group. CONCLUSIONS: Among patients with psychosis, having a diagnosis of schizophrenia and being male increase the likelihood of special hospital admission. Suggestions that ethnic minority patients are much more likely to have engaged in serious violence and need high-security placement were not borne out. PMID- 11925360 TI - Prader-Willi syndrome, compulsive and ritualistic behaviours: the first population-based survey. AB - BACKGROUND: Obsessive-compulsive disorder has been reported in association with Prader-Willi syndrome. AIMS: To report the nature and prevalence of compulsive and similar symptoms associated with Prader-Willi syndrome in a population ascertained as completely as possible. METHOD: Attempted complete ascertainment of people with Prader-Willi syndrome in eight English counties. Administration of standardised rating scales and a structured interview. Comparison with people with learning disability and high body mass indices. RESULTS: Prader-Willi syndrome was associated with high rates of ritualistic behaviours, such as the need to ask or to tell something, insistence on routines, hoarding and ordering objects and repetitive actions and speech, compared with the control group, and was negatively correlated with IQ and socialisation age. Typical obsessive compulsive symptoms, such as checking, counting and cleaning compulsions or obsessional thoughts, were not found. CONCLUSIONS: Ritualistic and compulsive behaviours occur more frequently in association with Prader-Willi syndrome than among people with intellectual disability and significant obesity. PMID- 11925361 TI - Dissociation and post-traumatic stress disorder: two prospective studies of road traffic accident survivors. AB - BACKGROUND: Dissociative symptoms during trauma predict post-traumatic stress disorder (PTSD), but they are often transient. It is controversial whether they predict chronic PTSD over and above what can be predicted from other post-trauma symptoms. AIMS: To investigate prospectively the relationship between dissociative symptoms before, during and after a trauma and other psychological predictors, and chronic PTSD. METHOD: Two samples of 27 and 176 road traffic accident survivors were recruited. Patients were assessed shortly after the accident and followed at intervals over the next 6 months. Assessments included measures of dissociation, memory fragmentation, data-driven processing, rumination and PTSD symptoms. RESULTS: All measures of dissociation, particularly persistent dissociation 4 weeks after the accident, predicted chronic PTSD severity at 6 months. Dissociative symptoms predicted subsequent PTSD over and above the other PTSD symptom clusters. Memory fragmentation and data-driven processing also predicted PTSD. Rumination about the accident was among the strongest predictors of subsequent PTSD symptoms. CONCLUSIONS: Persistent dissociation and rumination 4 weeks after trauma are more useful in identifying those patients who are likely to develop chronic PTSD than initial reactions. PMID- 11925362 TI - Accessibility of health and social services to immigrant elders: the Islington Study. AB - BACKGROUND: Numbers of immigrant elders are increasing and it is unclear whether they can access services. AIMS: To examine service utilisation of older immigrants compared with their UK-born counterparts and relate it to health difficulties. Method Cross-sectional study in inner London measuring service use, mental health and disability. RESULTS: A total of 1085 people aged > or = 65 years were interviewed. Independent predictors of contact with a general practitioner included being born in Cyprus. Cypriots were the only immigrant population to report significantly more somatic symptoms than those born in the UK (P=0.005). Africans and Caribbeans used day care and other social services most frequently. CONCLUSIONS: Immigrants could access services. Africans and Caribbeans appear to have poorer physical health and thus have greater contact with services. Cypriots who experience depression may present with prominent somatic symptoms. This is likely to be due to a different idiom of distress. PMID- 11925363 TI - War pensions (1900-1945): changing models of psychological understanding. AB - BACKGROUND: War pensions are used to examine different models of psychological understanding. The First World War is said to have been the first conflict for which pensions were widely granted for psychological disorders as distinct from functional, somatic syndromes. In 1939 official attitudes hardened and it is commonly stated that few pensions were awarded for post-combat syndromes. AIMS: To re-evaluate the recognition of psychiatric disorders by the war pension authorities. METHOD: Official statistics were compared with samples of war pension files from the Boer War and the First and Second World Wars. RESULTS: Official reports tended to overestimate the number of awards. Although government figures suggested that the proportion of neurological and psychiatric pensions was higher after the Second World War, our analysis suggests that the rates may not have been significantly different. CONCLUSIONS: The acceptance of psychological disorders was a response to cultural shifts, advances in psychiatric knowledge and the exigencies of war. Changing explanations were both a consequence of these forces and themselves agents of change. PMID- 11925364 TI - The case for cothymia: an open verdict? PMID- 11925365 TI - The case for cothymia: an open verdict? PMID- 11925366 TI - Cross-cultural training in psychiatry. PMID- 11925367 TI - Substance misuse among people with schizophrenia: similarities and differences between the UK and France. PMID- 11925370 TI - The ovarian endometrioma: why is it so poorly managed? Indicators from an anonymous survey. AB - As a tertiary referral centre for women with severe endometriosis we see a large number of patients who have endometriotic cysts, and many of these patients have been operated on by gynaecologists elsewhere in the UK. We have been surprised by the variety of approaches to their management. In order to establish the current clinical practices of endometrioma management amongst gynaecologists in the UK, we have carried out an anonymous survey. There appears to be an equal preference for open versus endoscopic surgery, even though there is considerable evidence suggesting that endoscopic surgery is superior. However, the management of endometriomas varies significantly between the two groups. In this article, we have presented the results of our survey and used them as the basis for a debate on the management of endometriomas. PMID- 11925371 TI - Ooplasmic donation in humans: the potential for epigenic modifications. AB - Ooplasm donation, wherein ooplasm is transferred from a donor oocyte to a recipient oocyte in an effort to increase embryo viability, has been applied in the human, with resulting pregnancies and births. Neither the safety nor efficacy of this method has been adequately investigated. Mitochondrial heteroplasmy in the blood of children conceived using ooplasm donation has recently been described. A follow-up study of children born following the use of this technique primarily focused on the presence of mitochondria from the donor oocyte highlighting possible problems due to mitochondrial heteroplasmy. Other effects related to epigenetic events may also arise, but have not been addressed. Studies using inbred mouse strains reveal that genetically diverse ooplasms can impose diverse epigenetic modifications on parental genomes. Incompatibilities produced by combining maternal genome and ooplasm from different genotypes leads to defects in gene expression and development. Such defects can be heritable and observed in the next generation. Given the potential for epigenetic modifications to arise following ooplasm donation, the safety and efficacy of this method need to be evaluated in a suitable animal model. PMID- 11925372 TI - Should patients with polycystic ovarian syndrome be treated with metformin? A note of cautious optimism. AB - Hyperinsulinaemia has proved to be a key link in the enigmatic generation of the symptoms of polycystic ovarian syndrome (PCOS), i.e. anovulatory infertility and the skin stigmata induced by hyperandrogenism. Regression of these symptoms may be achieved by reducing the hyperinsulinaemia. As obesity exaggerates the expression of the symptoms induced by hyperinsulinaemia, a low calorie diet and lifestyle change resulting in loss of weight for obese women with PCOS is capable of reversing these symptoms. Insulin-sensitizing agents, predominantly metformin, have been examined for their ability, in all patients with PCOS, to achieve similar beneficial changes to those induced by loss of weight in the obese. While the scientific value of many of these studies is questionable and solid evidence of efficiency and safety is not complete, the honourable intent of lowering high insulin levels in this way prompts the bottom line of this debate to strike a note of cautious optimism that insulin-sensitizing agents will be of some clinical usefulness both in the short-term aiding of infertility treatment and, possibly, in the prevention of the long-term sequelae for this troublesome and very prevalent condition. PMID- 11925373 TI - From HMG through purified urinary FSH preparations to recombinant FSH: a substitution study. AB - Drugs produced through the use of recombinant DNA techniques have become an integral part of medical practice. Before recombinant FSH (rFSH) was introduced in 1996, FSH purified from the urine of postmenopausal women had been commercially available since the 1960s. We analysed the diffusion and the substitution patterns of the different FSH preparations in The Netherlands. The fact that rFSH preparations have batch-to-batch consistency, are free from urinary protein contaminants and have the potential to be produced in limitless quantities, is advantageous. The question whether newer, more pure FSH products are beneficial from the clinical perspective, has not been settled beyond reasonable doubt. The price of rFSH is three times as high as the price of the former FSH preparations. Due to the introduction of rFSH, total FSH expenditures have grown from 5.0 million Euros in 1995, to 26.8 million Euros in 2000, while the volume increased by <100%. Both the pharmaceutical companies and purchasers (government, insurers) have influenced the patterns of substitution of existing FSH products by biotech equivalents. In general, the risk of increasing pharmaceutical costs without clear clinical benefits has to be set against the risk of strangling innovations. Therefore, a continuous process of technology assessment is necessary. PMID- 11925374 TI - The diaphragm with and without spermicide for contraception: a Cochrane review. AB - BACKGROUND: The diaphragm is usually used with a spermicide. However, some practitioners have suggested that spermicides offer no additional contraceptive protection and have advocated alternative guidelines for the use of diaphragms. The objective of this review was to compare the effectiveness, safety and acceptability of the diaphragm with and without spermicide. METHODS AND RESULTS: We searched Medline, Embase, Popline, CINAHL, the Cochrane Controlled Trials Register and reference lists of relevant articles. In addition, we contacted experts in the field to identify unpublished studies. Randomized controlled trials comparing women of reproductive age using the diaphragm with and without spermicide as the sole contraceptive method that reported clinical outcomes were selected. Two reviewers independently extracted data on outcomes and trial characteristics and any discrepancies were resolved by consensus or by consultation with the third reviewer. The results of the one identified study are presented descriptively. We identified only one study. No significant difference was found in the pregnancy rates (with typical use or consistent use) or discontinuation rates between the diaphragm-with-spermicide and diaphragm-without spermicide groups. There was a trend towards higher pregnancy rates in the diaphragm-without-spermicide group. However, this study failed to recruit the planned number of participants and was consequently underpowered. CONCLUSIONS: As only one underpowered study was identified, we cannot distinguish between the contraceptive effectiveness of the diaphragm with and without spermicide. We cannot draw any conclusion at this point; further research is needed. PMID- 11925375 TI - Biphasic versus monophasic oral contraceptives for contraception: a Cochrane review. AB - BACKGROUND: Side-effects caused by oral contraceptives discourage compliance with and continuation of oral contraceptives. Three approaches have been used to decrease these adverse effects: reduction of steroid dose, development of new steroids, and new formulas and schedules of administration. The third strategy led to the biphasic oral contraceptive pill. We compared biphasic oral contraceptives with monophasic oral contraceptives in terms of efficacy, cycle control and discontinuation due to side-effects. Our a priori hypotheses were: (i) biphasic oral contraceptives are less effective in preventing pregnancy than monophasic oral contraceptives, and (ii) biphasic oral contraceptives cause more side-effects, give poorer cycle control and have lower continuation rates. METHODS: We searched computerized databases Medline, Embase, Popline and the Cochrane Controlled Trial Register. Additionally, we searched the reference lists of all potentially relevant articles and book chapters. We also contacted the authors of relevant studies and pharmaceutical companies in Europe and the USA. We included randomized controlled trials comparing any biphasic oral contraceptive with any monophasic oral contraceptive when used to prevent pregnancy. We examined the studies found during the various literature searches for possible inclusion and assessed their methodological quality using the Cochrane guidelines. We contacted the authors of all included studies and of possibly randomized studies for supplementary information about the study methods and outcomes. We entered the data in RevMan 3.1, imported the data into RevMan 4.1, and calculated Peto odds ratios for the incidence of intermenstrual bleeding, absence of withdrawal bleeding and study discontinuation due to intermenstrual bleeding. RESULTS: Only one trial of limited quality compared a biphasic and monophasic preparation. This trial examined 533 user cycles of a biphasic pill (norethindrone 500 microg/ethinyl estradiol 35 microg for 10 days, followed by norethindrone 1000 microg/ethinyl estradiol 35 microg for 11 days) and 481 user cycles of a monophasic contraceptive pill (norethindrone acetate 1500 microg/ethinyl estradiol 30 microg daily). The study found no significant differences in intermenstrual bleeding, amenorrhoea and study discontinuation due to intermenstrual bleeding between the biphasic and monophasic oral contraceptive pills. CONCLUSIONS: Conclusions are limited by the identification of only one trial, the methodological shortcomings of that trial and the absence of data on accidental pregnancies. However, the trial found no important differences in bleeding patterns between the biphasic and monophasic preparations studied. Since no clear rationale exists for biphasic pills and since extensive evidence is available for monophasic pills, the latter are preferred. PMID- 11925376 TI - GnRH antagonist in assisted reproduction: a Cochrane review. AB - BACKGROUND: In the present systematic review, we wished to compare the efficacy of gonadotrophin-releasing hormone (GnRH) antagonist and GnRH agonist administration for controlled ovarian hyperstimulation in assisted conception. METHODS: Five randomized controlled trials fulfilled the inclusion criteria. In four studies, the multiple low-dose (0.25 mg) antagonist regimen was applied and, in one study, the single high-dose (3 mg) antagonist regimen was investigated. In all trials, reference treatment included a long protocol of GnRH agonist (buserelin, leuprorelin or triptorelin) starting in the mid-luteal phase of the preceding cycle. RESULTS: In comparison with the long protocol of GnRH agonist, the overall odds ratio for the prevention of premature LH surges was 1.76 [95% confidence interval (CI) 0.75-4.16], which was not statistically significant. There were significantly fewer clinical pregnancies in those treated with GnRH antagonists (OR 0.79; 95% CI 0.63-0.99). There was no statistically significant reduction in incidence of severe ovarian hyperstimulation syndrome between the two regimens (relative risk 0.51; OR 0.79; 95% CI 0.22-1.18). CONCLUSIONS: We concluded that the fixed GnRH antagonist protocol is a short and simple protocol with good clinical outcome, but the lower pregnancy rate compared with the GnRH agonist long protocol and the non-significant difference between both protocols regarding prevention of premature LH surge and prevention of severe ovarian hyperstimulation syndrome necessitates counselling subfertile couples before recommending change from GnRH agonist to antagonist. The clinical outcome may be further improved by developing more flexible antagonist regimens, taking into account individual patient characteristics. PMID- 11925377 TI - Morphometric and cytogenetic characteristics of testicular germ cells and Sertoli cell secretory function in men with non-mosaic Klinefelter's syndrome. AB - BACKGROUND: Klinefelter's syndrome is the most frequent chromosomal abnormality in infertile men. In this study, the chromosomes of round spermatids and spermatogonia/primary spermatocytes from men with non-mosaic Klinefelter's syndrome were examined, together with the Sertoli cell secretory function and sperm morphometry. METHODS: Twenty-four men with non-mosaic Klinefelter's syndrome and nine men with obstructive azoospermia underwent therapeutic testicular biopsy. When spermatozoa in the final filtrate were present, they were processed for sperm morphometry or ICSI. Sperm morphometry was evaluated by the maximal length and width of the sperm head, the length of the midpiece and the ratio of the acrosomal region to the total surface area of the head. When round spermatids were present, they were processed for fluorescent in-situ hybridization (FISH). FISH was also applied to fragments of seminiferous tubules. Sertoli cell secretory function was measured by the amount of androgen binding protein (ABP) secreted in vitro. RESULTS: More than 93% of the evaluated round spermatids were normal. The proportions of 24,XY and of 24,XX round spermatids to the total number were significantly larger in men with Klinefelter's syndrome than in obstructed azoospermic men. Men with Klinefelter's syndrome who had spermatozoa in their testicular tissue (n = 12) were positive for both 46,XY and 47,XXY spermatogonia in their seminiferous tubules. In contrast, men with Klinefelter's syndrome without spermatozoa in their testicular tissue (n = 12) were positive for 47,XXY spermatogonia but negative for 46,XY spermatogonia in their seminiferous tubules. ABP profiles were significantly smaller in men with Klinefelter's syndrome who were negative for spermatozoa compared with men who were positive. Four pregnancies were achieved and five healthy babies were born. CONCLUSIONS: This study suggests that few 46,XXY spermatogonia undergo meiosis with an XX pairing and a Y univalent type of pairing. Hyperhaploid round spermatids (24,XY and 24,XX) may be produced by meiosis of 47,XXY spermatogonia. Men with Klinefelter's syndrome who are negative for testicular spermatozoa have a greater degree of Sertoli cell secretory dysfunction compared with men with Klinefelter's syndrome who are positive for spermatozoa. There are several defects in sperm morphometry with functional significance in men with Klinefelter's syndrome. PMID- 11925378 TI - Association of the CYP17 gene and CYP19 gene polymorphisms with risk of endometriosis in Japanese women. AB - BACKGROUND: To investigate whether polymorphisms of CYP17 and CYP19 genes are associated with the risk of endometriosis, we analysed the frequency and distribution of a single nucleotide polymorphism at the 5' untranslated region of the CYP17 gene, and a tetranucleotide (TTTA) tandem repeat polymorphism and a 3 bp insertion (I)/deletion (D) polymorphism in intron 4 of the CYP19 gene. METHODS: We studied 140 patients with endometriosis, 67 with adenomyosis and/or leiomyomas and 177 healthy control women. RESULTS: The distribution of the genotypes of CYP17 and alleles of the TTTA repeat polymorphism of CYP19 were not significantly different between the groups. In contrast, an increased frequency of the D/D genotype was observed in the endometriosis group as compared with the control group (D/D genotype versus I/I plus I/D genotypes; corrected P = 0.024). This was more evident in the endometriosis subgroups with chocolate cysts (corrected P = 0.043) and at severe clinical stages (corrected P = 0.035). CONCLUSIONS: The results suggest that the 3 bp I/D polymorphism of the CYP19 gene may be weakly associated with the susceptibility of endometriosis in a Japanese population. PMID- 11925379 TI - A human homologue of mouse Mater, a maternal effect gene essential for early embryonic development. AB - BACKGROUND: Mater is a maternal effect gene required for early embryonic development in mice, and its protein serves as an autoantigen in a mouse model of autoimmune premature ovarian failure. METHODS: Human MATER cDNA was cloned by PCR techniques. The mRNA and protein were determined using hybridization and immunodetection respectively. The cDNA and protein sequences were analysed using bioinformatics software. RESULTS: Human MATER gene spans a approximately 63 kbp DNA at chromosome 19 and is composed of 15 exons and 14 introns. Expression of its mRNA (approximately 4.2 kb) is restricted to the oocytes. Human MATER cDNA (3885 nt) shows an open reading frame (3600 nt) encoding a polypeptide chain composed of 1200 residues with a predicted molecular mass of 134 236 Da. MATER protein (approximately 134 kDa) was detected in human oocytes. The human and mouse cDNA share 67% homology while their deduced polypeptide chains have 53% identity of amino acids. Also, their protein structures have a number of similar features. CONCLUSIONS: The human MATER and mouse Mater genes and proteins are conserved. Characterization of the human MATER and its protein provides a basis for investigating their clinical implications in autoimmune premature ovarian failure and infertility in women. PMID- 11925380 TI - Dizygotic twin boys born after ICSI with maternal meiosis I-derived free trisomy 21 in the first and multiple congenital anomalies in the second: chance or common aetiology? AB - We report on a pair of dizygotic twin boys born after ICSI. One twin was affected with maternal meiosis I-derived free trisomy 21. The other had multiple congenital malformations including a complex heart defect and oesophageal atresia. The advanced maternal age of 37 years predisposed for chromosome 21 meiosis I non-disjunction in twin A. Each of the multiple congenital anomalies in twin B has been described in trisomy 21. However, due to dizygosity demonstrated by a panel of molecular markers mapped on chromosome 21 as well as the results of investigations with 16 short tandem repeat markers localized on various other chromosomes, low level mosaicism or chimerism for this aneuploidy in twin B is unlikely. In addition, the twinning process, which by itself is associated with an increased rate of congenital malformations particularly affecting heart and oesophagus, might be responsible for the multiple congenital anomalies in twin B. Thus, in agreement with the results of several population-based studies from the literature, it appears unlikely that the micromanipulation of ICSI is causally responsible for the different anomalies found in these two boys. PMID- 11925381 TI - Induced apoptosis and expression of related proteins in granulosa cells from women undergoing IVF: a preliminary study. AB - BACKGROUND: Because apoptosis may be involved in the outcome of IVF, the expression of pro- and anti-apoptosis proteins in a model of induced granulosa cell (GC) apoptosis was evaluated in 25 women with normal FSH levels undergoing IVF. METHODS: After 1 day of culture, apoptosis was induced with interferon (IFN) gamma (200 IU/ml), followed 24 h later by an agonistic anti-Fas antibody (0.5 microg/ml). On day 3, apoptotic GC, identified by chromatin condensation and/or nuclear fragmentation after DAPI staining, were counted among 1000 cells in randomly chosen fields under UV microscopy, and enabled allocation of women into two groups with either low (group 1) or high (group 2) percentages of apoptosis (11.6 +/- 4.8 and 59.5 +/- 14.8% respectively; P < 0.001). Immunoblotting was used to evaluate the following in proteins: poly (ADP-ribose) polymerase (PARP), caspases 8 and 3, Bcl-2, heat shock protein (HSP) 70, Bax, Bak and Stat-1 (a protein known to be inducible by IFN-gamma). RESULTS: Based on densitometric analysis of immunoblots, the PARP 116 kDa bands were respectively 4.3- and 33.3 fold lower for treated groups 1 and 2. Caspase 8, caspase 3 and HSP70 were expressed slightly less in treated group 2 than treated group 1. Densitometric analysis of bands corresponding to Bcl-2 showed respectively for treated groups 1 and 2, 3.2- and 2.5-fold decreases. Bak expression was similar in both control groups, and comparably lower in the two treated groups. With regard to Stat-1, densitometry showed 3.3- and 1.3-fold increases respectively in treated groups 1 and 2. CONCLUSIONS: These results suggested that Fas-mediated apoptosis of GC is accompanied by significant changes in proteins acting in apoptosis, and that this type of programmed cell death might play a potential prognostic role for women undergoing IVF. PMID- 11925382 TI - Smoking induces oxidative stress inside the Graafian follicle. AB - BACKGROUND: A growing body of evidence indicates that pro-oxidant/antioxidant balance inside ovarian follicles plays an important role in folliculogenesis. Over 20% of women of reproductive age in Europe and the USA regularly smoke cigarettes. The impact of tobacco smoking on the intrafollicular markers of oxidative stress has not been fully elucidated. The objective of the present study was to test the hypothesis that cigarette smoking affects the intrafollicular redox milieu. METHODS: In follicular fluid samples originating from 108 IVF patients, lipid peroxidation was assessed by the thiobarbituric reactive substances method and total antioxidative capacity was quantified by the luminol enhanced chemiluminescence method. The level of patients' exposure to the cigarette smoke was evaluated by measuring the follicular fluid cotinine concentration by means of radioimmunoassay. RESULTS: Intrafollicular exposure to cigarette smoke metabolites was associated with a significant increase in follicular lipid peroxidation intensity (P < 0.001), which was accompanied by a significant decrease in the local antioxidative potential (P = 0.004). CONCLUSION: The results indicate that active smoking affects the pro oxidant/antioxidant balance inside the pre-ovulatory ovarian follicle by inducing intrafollicular oxidative stress. This provides another possible explanation for impaired folliculogenesis in female smokers. PMID- 11925383 TI - Comparison of day 3 FSH serum values as determined by six different immunoassays. AB - BACKGROUND: The accurate assessment of FSH concentration is important for evaluating ovarian function prior to IVF. However, a number of different assay techniques are currently in use, leading to inconsistencies in the hormone data being reported. To address this problem, we measured FSH concentration using a number of commercially available systems. METHODS: Day 3 serum FSH levels were measured in 215 healthy fertile women using six different immunoassays: Coatria (125)I (Bio-Merieux), ACS-180 (Bayer Diagnostics), Advia-Centaur (Bayer Diagnostics), Vitros ECi (Ortho-Clinical Diagnostics), Architect i2000 (Abbott) and Elecsys 2010 (Roche Diagnostics). RESULTS: According to the immunoassay, means +/- SD of FSH concentrations were: 6.5 +/- 2.2 mIU/ml for Coatria (125)I, 6.8 +/- 2.7 mIU/ml for Advia-Centaur, 6.7 +/- 3.0 mIU/ml for Vitros ECi, 7.6 +/- 3.0 mIU/ml for ACS-180, 8.2 +/- 3.3 mIU/ml for Architect i2000 and 8.8 +/- 3.0 mIU/ml for Elecsys 2010. CONCLUSION: Day 3 FSH values determined by six different immunoassays were significantly different (P < 0.01, paired t-test). Physicians must take care when interpreting results from different clinical laboratories. PMID- 11925384 TI - Hospital care utilization of infants born after IVF. AB - BACKGROUND: Infants born after IVF are often twins, and singleton IVF babies have an increased risk for preterm birth. Both conditions are likely to increase morbidity. We examined the frequency and duration of hospitalization required by babies born after IVF, and compared this information with all infants born in Sweden during the same time period. METHODS: We used a nationwide registration of IVF pregnancies from 1984 to 1997 and a nationwide register of all in-patient care up to the end of 1998. We identified 9056 live born infants after IVF treatment and compared them with 1 417 166 non-IVF live born infants. RESULTS: The highest odds ratio (OR approximately 3) was seen for neonatal hospitalization, but an increased OR (1.2-1.3) was noted for children up to 6 years of age. The OR for being hospitalized after IVF was 1.8, but when the analysis was restricted to term infants it was 1.3 and this excess was then explainable by maternal subfertility. Statistically significant increased ORs were seen for hospitalization for cerebral palsy (1.7), epilepsy (1.5), congenital malformation (1.8) or tumour (1.6), but also for asthma (1.4) or any infection (1.4). When information from the Swedish Cancer Registry was used, no excess risk for childhood cancer was found. The average number of days spent in hospital by IVF and non-IVF children was 9.5 and 3.6 respectively. CONCLUSIONS: The increased hospitalization of IVF children is, to a large extent, due to the increased incidence of multiple births. Therefore, the increased costs associated with this may be reduced by the use of single embryo transfers, with the savings in health care costs being offset against the increased number of embryo transfer cycles required to maintain the pregnancy rate. PMID- 11925386 TI - Cumulative delivery rates after ICSI in women aged >37 years. AB - BACKGROUND: Female patients aged >37 years have a poor prognosis after ICSI. To determine the cumulative delivery rates in these women by life-table analysis, 228 patients aged >37 years who had undergone a total of 437 ICSI cycles were analysed retrospectively. METHODS: Only cycles in which fresh ejaculated sperm was used, and in which at least one oocyte was micro-injected, were analysed. The main outcome measure was cumulative rate of deliveries. Any delivery after 25 weeks gestation was included in the study. RESULTS: In women aged 38-39 years, the real cumulative delivery rate after two cycles was 21%, while the expected delivery rate was 26%. In patients aged 40-43 years, the real and expected cumulative delivery rates were 12 and 17% respectively after three cycles, when they reached a plateau. There was only one delivery in the age group >43 years, which consisted of 26 patients with 66 cycles. In women aged >37 years, an expected cumulative delivery rate of 30% may be obtained at the end of the fourth cycle. Women aged >43 years do not have a realistic chance of achieving a delivery with their own oocytes. CONCLUSIONS: This life-table analysis provides a means by which to counsel couples about their chances of achieving a delivery by ICSI at an age >37 years. PMID- 11925385 TI - Progesterone serum levels during the follicular phase of the menstrual cycle originate from the crosstalk between the ovaries and the adrenal cortex. AB - BACKGROUND: The preovulatory rise of progesterone is important for ovulation, but both its regulation and its origin are controversial. Three experiments were performed to determine whether follicular phase progesterone arises from the ovary, the adrenal cortex or both. METHODS: The first study was performed in patients scheduled for assisted reproduction, who received a long-acting GnRH agonist either during intake of an oral contraceptive or during the luteal phase of an otherwise untreated menstrual cycle. The second study was also performed during down-regulation with a GnRH agonist: some patients with elevated progesterone levels received dexamethasone (DXM). Others with similarly elevated basal progesterone levels and those with low progesterone levels were not treated with DXM and served as controls. Finally, adrenocorticotrophic hormone (ACTH) tests were performed in normocyclic volunteers both during early and late follicular phase and during intake of a contraceptive pill. RESULTS: During the suppression of endogenous gonadotrophin secretion progesterone levels rose after the administration of ACTH, but not of GnRH. DXM did not prevent the preovulatory rise of the serum progesterone concentration. The ACTH-stimulated concentration of progesterone and of 17alpha-hydroxyprogesterone were significantly reduced during intake of ethinyl estradiol. CONCLUSIONS: Progesterone arises in the adrenal cortex during most of the follicular phase, whereby its function is modulated by an unknown ovarian factor, which is suppressed by ethinyl estradiol. The source of progesterone shifts towards the ovaries prior to ovulation. PMID- 11925387 TI - The effect of various infertility treatments on the risk of preterm birth. AB - BACKGROUND: The high incidence of preterm birth (<37 weeks gestation) is a major concern in assisted reproductive technology. The objective of this study was to compare the risk of preterm birth from singleton pregnancies following either low technology treatment (intrauterine insemination and donor insemination) or high technology treatment (IVF, ICSI and gamete intra-Fallopian transfer) with that of naturally conceived pregnancies. METHODS: Three cohorts of pregnancies resulting from either low or high technology treatment or from natural conception were included in the study. A number of potential risk factors were adjusted for. RESULTS: The incidence of very preterm birth (<32 weeks of gestation) was not significantly increased in the low technology treatment group (1.0 versus 1.3% in controls) but was significantly higher in the high technology treatment group (5.2%, P < 0.001). In spontaneous, elective Caesarean section (CS) and induced delivery onset, the risk of preterm birth increased gradually from the controls to the low technology treatment group to the high technology treatment group, while for an emergency CS the risk of preterm birth was very high in both treatment groups. CONCLUSIONS: The overall incidence of preterm birth increased significantly from the controls to the low technology treatment group and to the high technology treatment group. Logistic regression analysis showed that younger and older age, previous perinatal death and emergency CS were associated with an increased risk, while a previous live birth reduced the risk. The length of the infertile period did not seem to affect the risk in any of the treatment groups. PMID- 11925388 TI - Predictors of IVF outcome by three-dimensional ultrasound. AB - BACKGROUND: Our study was designed to evaluate whether ovarian antral follicle number, ovarian volume, stromal area and ovarian stromal blood flow are predictive of ovarian response and IVF outcome. A total of 56 women with normal basal serum FSH concentrations who had no history of ovarian surgery, no ovarian and/or uterine pathology, were non-smokers and undergoing their first IVF cycle using a standard long GnRH agonist protocol were examined. METHODS: Total ovarian antral follicle number, total ovarian volume, total stromal area and mean flow index (FI) of the ovarian stromal blood flow were determined by three-dimensional (3D) and power Doppler ultrasound after pituitary suppression. Pretreatment 3D ultrasound ovarian measurements were compared with subsequent ovulation induction parameters [peak estradiol (E2) on HCG administration day and number of oocytes] and cycle outcome (fertilization and pregnancy rates). RESULTS: The total antral follicle number achieved the best predictive value for favourable IVF outcome, followed by ovarian stromal FI, peak E2 on HCG administration day, total ovarian volume, total ovarian stromal area and age. Using these six parameters, we were able to predict a favourable IVF outcome in 50% (11/22) of patients and poor outcome in 85% (29/34) of patients. CONCLUSIONS: Three-dimensional ultrasound facilitates determination of the antral follicle number, ovarian volume calculation, evaluation of the ovarian stroma and analysis of the intensity of ovarian stromal blood flow in a short time without increasing the patient's discomfort. PMID- 11925389 TI - The use of intrauterine insemination in Australia and New Zealand. AB - BACKGROUND: There is good evidence in the literature in favour of intrauterine insemination (IUI) as the most cost-effective treatment for unexplained and moderate male factor subfertility. However there is no published data on whether this evidence is being translated into clinical practice. METHODS: We identified fertility centres within Australia and New Zealand registered with the Reproductive Technology Accreditation Committee of the Fertility Society of Australasia. Thirty-seven of these units were then sent a postal survey to establish current clinical practice. RESULTS: Nearly a third of centres promote IVF as first-line treatment even in the presence of patent tubes and normal semen while, when semen parameters are reduced, IUI is rarely considered. One in five (20%) units remain unconvinced of the cost-effectiveness of IUI. When IUI is used, it is virtually always combined with ovarian stimulation with marginally more units using clomiphene citrate than gonadotrophins. CONCLUSIONS: Although it may take relatively more treatment cycles to achieve pregnancy, there are considerable advantages to the patient in terms of risk/benefit ratio and financial cost associated with IUI compared with IVF. In the current climate of evidence-based medicine, as clinicians we are obliged to translate this into our practice. It appears from our survey that in many units this is not happening. PMID- 11925390 TI - Prolactin and growth hormone secretion after thyrotrophin-releasing hormone infusion and dopaminergic (DA2) blockade in infertile patients with minimal/mild endometriosis. AB - BACKGROUND: The origin of infertility in patients with endometriosis without tubal occlusion has not yet been clearly defined. Several reports show an abnormal pituitary-ovarian axis in this group of patients. Moreover, prolactin (PRL) and growth hormone (GH) secretion is closely related to reproductive status. This study aimed to evaluate PRL and GH secretion after metoclopramide and thyrotrophin-releasing hormone (TRH) infusion in infertile patients with minimal/mild endometriosis. METHODS: A total of 64 women participated in the study: 33 fertile patients without endometriosis; 10 fertile patients with minimal/mild endometriosis; and 21 infertile patients with minimal/mild endometriosis. TRH or metoclopramide was administered randomly in two sequential menstrual cycles (cycle days 3-5). Serum PRL and GH secretion before and after dopaminergic type 2 (DA2) receptor blockade and TRH were compared. RESULTS: Higher serum PRL levels were observed in patients with endometriosis at baseline and after 15 and 30 min of TRH administration. Also, infertile patients with endometriosis had lower serum estradiol levels than fertile patients. Moreover, the dopaminergic blockade did not result in abnormal PRL or GH secretion. CONCLUSIONS: Decreased serum estradiol levels and altered PRL secretion after TRH administration in infertile patients with minimal/mild endometriosis are related to ovulatory dysfunction and infertility in this group of patients without tubal occlusion. PMID- 11925391 TI - Ovarian hyperstimulation syndrome complicated by peritonitis due to perforated appendicitis. AB - We describe a case of ovarian hyperstimulation syndrome (OHSS) complicated by peritonitis due to perforated appendicitis. A 29-year-old woman presented with abdominal distension after ovarian stimulation with HMG followed by ovulation induction with HCG. Massive ascites with swollen ovaries was observed on ultrasound, and she was admitted on the diagnosis of OHSS. Daily infusion of serum albumin and low dose dopamine failed to increase her urine output and her abdominal symptoms became increasingly deteriorated after her urine pregnancy test turned out to be positive. Paracentesis performed for alleviation of her abdominal distension revealed infected, foul-smelling fluid. An emergency laparotomy was performed, and the definite diagnosis was made as panperitonitis due to perforated appendicitis with right tubal pregnancy. Appendectomy, right tubectomy and vigorous irrigation with drainage were performed. The case implies that OHSS might not only mask typical manifestations of appendicitis, but could also compromise concurrent intraperitoneal infection. PMID- 11925393 TI - Serum inhibin B cannot predict testicular sperm retrieval in patients with non obstructive azoospermia. AB - BACKGROUND: Serum inhibin B, a direct product of the Sertoli cells, may serve as a marker of spermatogenesis. The present retrospective study aimed at evaluating the predictive value of inhibin B for retrieving testicular sperm in non obstructive azoospermic men. METHODS: The serum inhibin B concentration before sperm retrieval was reviewed in 185 non-obstructive azoospermic patients. RESULTS: Testicular sperm were successfully recovered in 92 of 185 patients (49.7%). The mean inhibin B concentration in these patients was 37.3 pg/ml. No sperm were found in 93 patients (50.3%), and the mean serum inhibin B concentration was 44.9 pg/ml. The discrimination between successful and unsuccessful sperm retrieval was analysed using the receiver operating characteristics (ROC) curve analysis. The best discriminating inhibin B concentration was 13.7 pg/ml (sensitivity 44.6%, specificity 63.4%) with an area under the ROC curve (AUC) of 0.51. Combining both serum FSH and inhibin B did not improve the predictive value: the AUC of inhibin B in men with a serum FSH concentration <25 and > or = 25 IU/l (being the best threshold value in the population studied) was respectively 0.53 and 0.50. The AUC of the inhibin B:FSH ratios was 0.55. CONCLUSIONS: This analysis shows that inhibin B, either alone or in combination with serum FSH, fails to predict the presence of sperm in men with non-obstructive azoospermia undergoing testicular sperm extraction. PMID- 11925392 TI - The significance of anti-Mullerian hormone concentration in seminal plasma for spermatogenesis. AB - BACKGROUND: The function of anti-Mullerian hormone (AMH) in seminal plasma in adulthood is uncertain. We examined the significance of seminal AMH for spermatogenesis. METHODS: We measured seminal concentrations of AMH in 39 oligozoospermic men (mean age +/- SD, 32.7 +/- 4.3 years) and 10 normal volunteers to examine the association of seminal AMH with spermatogenesis. The seminal concentrations of AMH in oligozoospermic men (149.3 +/- 254.0 pmol/l) were significantly lower than in normal men (249.0 +/- 167.7 pmol/l; P +/- 0.0337). Seminal AMH concentration correlated significantly with sperm concentration (r = 0.339, P = 0.0350) and mean testicular volume (r = 0.440, P = 0.246). The serum concentration of LH (r = - 0.365, P = 0.0241), but not FSH, testosterone or estradiol, correlated significantly with AMH concentration in seminal plasma. CONCLUSIONS: AMH in seminal plasma may be important for sperm production, and is a good marker for Sertoli cell development. PMID- 11925394 TI - Flow cytometric ploidy analysis of testicular biopsies from sperm-negative wet preparations. AB - BACKGROUND: The use of testicular sperm in assisted reproduction depends on the availability of sperm in wet preparations. It is not always possible to recover sperm from the testis, even with previous sperm-positive histopathological findings. The purpose of this study was to evaluate the sperm-negative wet preparation search results with flow cytometric ploidy analysis and histopathological examination. METHODS: Two pieces of testicular tissue were obtained from azoospermic patients to investigate the spermatogenic status of the testis, and to determine the presence of sperm through a wet preparation. The testicular tissue was shredded and then vortexed; the cellular suspension was then processed for a wet preparation sperm search, while the residual tissue was exposed to enzymatic digestion for flow cytometric ploidy analysis. RESULTS: A total of 38 patients had sperm-negative wet preparation results. Of those, six (16%) were shown to have haploid cells after flow cytometric analysis. Histopathological examination showed three samples with maturation arrest at the spermatid stage, and the other three at the spermatocyte stage. CONCLUSIONS: Flow cytometric ploidy analysis can be used to verify the results of a wet preparation sperm search when no sperm were detected. Flow cytometric ploidy analysis can also reveal the presence of spermatids when no sperm are available. PMID- 11925395 TI - Use of antisperm antibodies in differential display Western blotting to identify sperm proteins important in fertility. AB - BACKGROUND: Antisperm antibodies (ASA) may be an important cause of infertility but current tests for the detection of ASA have poor prognostic value. The inadequacy of current tests may reflect the inability of these tests to define the antigenic specificity of the sperm proteins with which the ASA react. Identification of the sperm proteins that ASA bind to is a necessary preliminary step to the development of more useful diagnostic tests for ASA. METHODS: A sensitive Western blotting technique was used to compare the antigenic specificities of ASA from men who were infertile (n = 6) with those who were fertile following vasectomy reversal (n = 3). Normal fertile men (n = 3) and infertile men with known ASA (n = 4) were also included in the analysis. RESULTS: All men, including the normal fertile controls, had ASA detectable in our system. Several sperm proteins were identified that react with ASA from infertile but not fertile men. Quantitative differences in the binding of ASA to some proteins were also demonstrated. Additionally, we demonstrated that normal motile sperm are coated with an antibody that appears to be bound to sperm by a non-antigenic mechanism. CONCLUSION: Sera from all men contained ASA, but clearly some of these did not cause infertility. Characterization of the proteins that are antigens for ASA from infertile but not fertile men may allow the development of more accurate tests for infertility-inducing ASA. The significance of immunoglobulin G coated on normal sperm remains to be determined. PMID- 11925396 TI - The spectrum of DNA damage in human sperm assessed by single cell gel electrophoresis (Comet assay) and its relationship to fertilization and embryo development. AB - BACKGROUND: The integrity of sperm DNA is important for the success of natural or assisted fertilization, as well as normal development of the embryo, fetus and child. ICSI, by bypassing sperm selection mechanisms, increases the risk of transmitting damaged DNA and the significance of this requires investigation. METHODS: DNA damage in sperm from an unselected group of 60 men undergoing IVF treatment was measured by single cell gel electrophoresis (Comet assay) and correlated with semen and treatment cycle parameters. RESULTS: Wide spectra of sperm DNA damage were found both within and between men but no specific subgroups were identified. Semen and treatment cycle parameters were not different in men grouped according to high or low sperm DNA damage. However, regression analysis showed that DNA damage was positively associated with age (29-44 years), abnormal sperm and motility and negatively associated with sperm concentration. In ICSI cycles DNA damage was positively associated with impairment of post-fertilization embryo cleavage. CONCLUSIONS: This study contributes to the evidence of DNA damage within sperm. High loads of DNA damage measured by the Comet assay were predictive of failure of embryo development after ICSI. As it is likely that sperm with DNA damage contributed to successful fertilization and in-vitro development, potential adverse effects remain to be clarified. PMID- 11925397 TI - Non-invasive amino acid turnover predicts human embryo developmental capacity. AB - BACKGROUND: IVF is limited by low success rates and a confounding high multiple birth rate contributing to prematurity, increased neonatal mortality and child handicap. These problems could be overcome if single embryos of known developmental competence could be selected for transfer on day 2/3 of development, but current methods, which rely on morphological appearance, are poor predictors of viability. METHODS: We have measured non-invasively the depletion/appearance (i.e. turnover) of a physiological mixture of 18 amino acids by single human embryos during in-vitro culture using high performance liquid chromatography. RESULTS: From the time of transfer (day 2/3), embryos with future competence to develop to the blastocyst stage (day 5/6) exhibit amino acid flux patterns distinct from those of embryos with similar morphological appearance which arrest. Significantly, the profiles of Ala, Arg, Gln, Met and Asn flux predict blastocyst potentiality at >95%. The amino acid most consistently depleted throughout development by those embryos which form blastocysts was leucine. Of the amino acids which were produced, the most striking was alanine, which appeared in increasing amounts throughout development. CONCLUSIONS: Non invasive amino acid profiling has the potential to select developmentally competent single embryos for transfer, thereby increasing the success rate and eliminating multiple births in IVF. PMID- 11925398 TI - Assessment of nuclear and cytoplasmic maturation in in-vitro matured human oocytes. AB - BACKGROUND: With improved prospects for the use of human oocyte in-vitro maturation in assisted reproductive technologies, the need to define more clearly the coordination of nuclear and cytoplasmic maturation has arisen. METHODS: Immunofluorescence and confocal microscopy were used to evaluate cell cycle dependent modifications in chromatin and microtubules in human germinal vesicle oocytes (n = 455) undergoing in-vitro maturation. RESULTS: Four distinct classes of germinal vesicle stage oocytes were identified based on the expression of G2/interphase characteristics, but, of these, only one class of oocytes was competent to complete meiotic progression to metaphase-II in vitro. The majority of germinal vesicle stage oocytes resumed meiosis within 6 h (88.9%) of culture and exhibited an accelerated pace of progression to metaphase-II (66.7%) over 24 h, but in general were unable to maintain meiotic arrest and defaulted into interphase within 24 h of polar body emission. Characterization of microtubule dynamics and chromatin phosphorylation demonstrates specific cell cycle deficiencies in in-vitro matured human oocytes. CONCLUSION: This work forms a basis for future studies aimed at optimizing nuclear and cytoplasmic maturation during in-vitro maturation. PMID- 11925399 TI - Follicular fluid markers of oocyte developmental potential. AB - BACKGROUND: Concentrations of certain substances in follicular fluid (FF) are related to fertilization outcome and early post-fertilization development. The study aim was to identify FF markers with which to predict embryo implantation potential. METHODS: Concentrations of selected hormones, cytokines and growth factors in individual FF samples obtained during assisted reproduction treatment were related with treatment outcomes. RESULTS: Mean concentrations of LH, growth hormone (GH), prolactin (PRL), 17beta-estradiol (E2) and insulin-like growth factor (IGF)-I were higher, and that of interleukin-1 (IL-1) was lower, in treatment attempts leading to a clinical pregnancy as compared with those in which no pregnancy was established. Concentrations of FSH, progesterone, tumour necrosis factor-alpha and IGF-II were similar in successful and unsuccessful attempts. In successful attempts, LH and GH levels were higher in those follicles from which oocytes giving rise to transferred embryos (i.e. embryos with best morphology and fastest cleavage rate) originated, as compared with other follicles from which a mature oocyte was recovered but was cryopreserved for later use. CONCLUSIONS: FF levels of LH, GH, PRL, E2, IGF-I and IL-1 may serve to analyse cases of repeated assisted reproduction failures and to assess effects of modifications of the ovarian stimulation protocol. PMID- 11925400 TI - Different perspectives of patients and health care professionals on the potential benefits and risks of blastocyst culture and multiple embryo transfer. AB - BACKGROUND: A trade-off exists between the risk of multiple pregnancy and prospects of pregnancy itself in assisted reproduction. Blastocyst culture and embryo transfer after approximately 5 days may be one method of reconciling this dilemma, although a controversial one. METHODS AND RESULTS: We presented a questionnaire to groups of patients, embryologists and clinicians to solicit views on the potential benefits and risks of blastocyst culture and multiple pregnancy. The results indicate that patients are more accepting of multiple pregnancy as a prospective outcome of treatment than those involved in their treatment, despite awareness of the risks. Our data tend to support a genuine difference in values on this point. We also sought views on the patient selection criteria and treatment protocols which should apply in a planned randomized controlled trial comparing blastocyst culture with cleaving embryo transfer (e.g. numbers of embryos to transfer, acceptable levels of risk of twin and triplet pregnancy, the proportion of patients who would be put off from entering the trial by the risk of no embryo transfer). These are presented and discussed with reference to their likely impact on trial recruitment, highlighting differences in perspective between patients and professionals. CONCLUSIONS: We conclude that there are differences among patients, embryologists and clinicians in their perceptions of the desirability of multiple pregnancy, their preferences in certain practical aspects of treatment such as embryo transfer numbers, and their ideas on blastocyst culture and its prospective outcomes and risks. PMID- 11925402 TI - The correlation of adhesions and peritoneal fluid cytokine concentrations: a pilot study. AB - BACKGROUND: Intra-abdominal adhesion formation and reformation after surgery is a significant cause of morbidity. The greatest problem after the surgical removal of adhesions is their reformation. We examined the concentrations of interleukin (IL)-1, IL-6 and tumour necrosis factor (TNF)-alpha in the peritoneal fluid throughout the 48 h post-operative period following adhesiolysis, and correlated the results to the extent of adhesion reformation. METHODS: Peritoneal fluid, collected from eight patients following laparoscopy and again at 12, 36 and 48 h after surgery, was analysed using enzyme-linked immunosorbent assay (IL-1 and IL 6) and bioassay (TNF-alpha). At 48 h, a second look laparoscopy was performed to inspect the pelvis for adhesion formation/reformation. RESULTS: Three patients had adhesion reformation >10% at 48 h after surgery. The mean adhesion score 48 h after adhesiolysis was 5 (range 0-17). The mean reduction in adhesion score was 88% (range 83-100%). Newly formed adhesions were filmy, relatively soft and avascular in nature. Adhesion reformation of >10% was associated with (i) high concentrations of IL-6 at 12 h (P < 0.01) and (ii) high concentrations of IL-1 at 48 h (P < 0.001). CONCLUSIONS: Results from this preliminary study suggest that future treatment strategies for adhesion prevention could be aimed at the control of cellular mediators in the peritoneal fluid during the initial adhesion formation period. PMID- 11925401 TI - A randomized, controlled pilot study of the safety and efficacy of 4% icodextrin solution in the reduction of adhesions following laparoscopic gynaecological surgery. AB - BACKGROUND: Adhesion-related readmissions are frequent sequelae to gynaecological surgery. Attempts to prevent adhesions by separating healing peritoneal surfaces include site-specific barriers and hydroflotation by instilled solutions. Rapid absorption limits the effectiveness of solutions such as Ringer's lactated saline (RLS). This pilot study assessed the safety, tolerability and preliminary effectiveness of a non-viscous, iso-osmolar solution of 4% icodextrin, an alpha 1,4 glucose polymer with prolonged intraperitoneal residence, in reducing adhesions after laparoscopic gynaecological surgery. METHODS: Women aged > or = 18 years, requiring laparoscopic adnexal surgery (n = 62), were entered into a randomized, open-label, assessor-blinded, multicentre study to compare 4% icodextrin with RLS. Treatments were coded in blocks of four with equal randomization to each group, and pre-allocated to consecutively numbered patients. At least 100 ml per 30 min was used for intra-operative lavage, with 1 l instilled post-operatively. Per protocol analysis included all eligible patients (n = 53); reformation analysis required one or more baseline adhesion (n = 42). Incidence, extent and severity of post-operative adhesions were assessed at second-look laparoscopy after 6-12 weeks. Procedures were video-taped for third party, blinded assessment. RESULTS: Safety and tolerability (laboratory variables, adverse events, clinical follow-up) were good with no difference between treatments. A shift analysis of incidence-ranked adhesions (n = 53) showed apparent improvements in more patients with icodextrin than RLS (37 versus 15%; not significant). Adhesion score reduction (n = 42) was more frequent in icodextrin- than RLS-treated patients: incidence (52 versus 32%), extent (52 versus 47%), and severity (65 versus 37%). Despite greater baseline adhesions, median reformation was less after icodextrin (24%) than RLS (60%). The pilot study group sizes were not powered for statistical significance. CONCLUSIONS: In this preliminary study, 4% icodextrin lavage plus instillation was well tolerated and reduced adhesion formation and reformation following laparoscopic gynaecological surgery. A Phase III pivotal study is currently in progress. PMID- 11925403 TI - Laparoscopic ovarian diathermy in women with polycystic ovarian syndrome: a retrospective study on the influence of the amount of energy used on the outcome. AB - BACKGROUND: Currently, there is an uncertainty about the optimum number of punctures to be applied at laparoscopic ovarian diathermy (LOD). This retrospective study was undertaken to investigate the dose-response relationship of LOD. METHODS: The hospital records of 161 women with polycystic ovarian syndrome who underwent LOD were reviewed and the clinical data before and after LOD were documented. Subjects were divided into six groups according to the number of punctures made in their ovaries as follows: group 1, two punctures per ovary; group 2, three punctures; group 3, four punctures; group 4, five punctures; group 5, six punctures and group 6, seven to 10 punctures. Contingency table analysis and analysis of variance were used to compare the outcomes of the different groups. RESULTS: The rates of ovulation, conception and restoration of menstrual regularity after LOD were significantly lower in group 1 compared with other groups. There were no significant differences in the success rates between the other groups. CONCLUSION: Two punctures per ovary are associated with poor results. Three punctures per ovary seem to represent the plateau dose. The application of seven or more punctures per ovary may result in excessive destruction to the ovary without any improvement of the results and should therefore be discouraged. PMID- 11925404 TI - Three-dimensional assessment of ultrasound features in women with clomiphene citrate-resistant polycystic ovarian syndrome (PCOS): ovarian stromal volume does not correlate with biochemical indices. AB - BACKGROUND: Polycystic ovarian syndrome (PCOS) is a heterogeneous endocrine disorder affecting women of reproductive age. The syndrome is characterized by a combination of polycystic ovarian morphology, clinical features and biochemical indices. The objective of this prospective observational study was to investigate the relationship between the total ovarian volume and the ovarian stromal volume, measured using a three-dimensional (3D) ultrasound scan, with biochemical indices of PCOS. METHODS: Twenty-three infertile women (mean age +/- SD: 31 +/- 3.1 years; range: 26-37) with clomiphene citrate-resistant PCOS were examined. Early follicular phase (days 2-4) serum FSH, LH and testosterone concentrations were measured following a withdrawal bleed. Total ovarian volume, stromal volume, pre antral follicle (<10 mm) number and total follicular volume were measured by 3D transvaginal ultrasound. RESULTS: Total ovarian volume, follicular volume and follicle number correlated positively with serum FSH and LH, but not testosterone, concentrations. Ovarian stromal volume correlated strongly with total ovarian volume only, but not with serum FSH, LH or testosterone concentrations. CONCLUSIONS: These findings demonstrate an association of the total ovarian volume, pre-antral follicle number and total follicular volume with some of the biochemical indices of PCOS, but no association was observed between ovarian stromal volume and these indices. PMID- 11925405 TI - An in-vivo study on placental transfer of naproxen in early human pregnancy. AB - BACKGROUND: Naproxen is one of the most common non-steroidal anti-inflammatory drugs used by women of reproductive age. Naproxen is known to be teratogenic in animals. The aim of this study was to investigate the placental transfer of naproxen in the first trimester of human pregnancy, and to determine the amount of the drug in different embryonic compartments. METHODS: Twenty-eight patients who requested surgical termination of pregnancy in the first trimester were given two oral 500 mg doses of naproxen before the surgical procedure. Four biological samples, maternal venous blood, coelomic fluid, amniotic fluid and fetal tissue, were collected from each patient for drug analyses by high performance liquid chromatography. RESULTS: Naproxen was detected in all samples. The mean (+/- SD) concentrations were 69.5 +/- 12.2 microg/ml, 6.4 +/- 2.4 microg/g, 1.85 +/- 1.03 microg/ml and 0.14 +/- 0.11 microg/ml in maternal serum, fetal tissue, coelomic fluid and amniotic fluid respectively. The mean amniotic fluid/maternal drug ratio and fetal/maternal drug ratio were 0.002 (range 0.0005-0.0064) and 0.092 (range 0.022-0.155) respectively. There was a positive correlation between the fetal drug concentration (r = 0.59, P = 0.001), amniotic fluid drug concentration (r = 0.47, P = 0.013), amniotic fluid/maternal ratio (r = 0.536, P = 0.003) and fetal/maternal ratio (r = 0.72, P < 0.001) with advancing gestational age. CONCLUSIONS: Although naproxen can cross the placenta readily in the first trimester of human pregnancy, only a small amount was present in fetal tissues. Since there is no information on whether this small amount of naproxen would be teratogenic or not, women of reproductive age who are taking naproxen regularly should be warned of the possible fetal side-effects. PMID- 11925406 TI - Reliability of serial urine HCG as a biomarker to detect early pregnancy loss. AB - BACKGROUND: To examine the reliability of HCG as a biomarker for early pregnancy loss, five experienced researchers independently assessed data from 153 menstrual cycles, determining whether each cycle represented 'no conception,' a 'continuing conception' or a 'conception lost.' METHODS: Urine samples were analysed by immunoradiometric assay using a combination of capture antibodies for the intact heterodimer (B109) and for an epitope common to the beta subunit and the beta core fragment (B204). For each cycle, HCG data were presented as graphs of daily assay results. Summary statistics for HCG assays from 46 women who had undergone bilateral tubal ligation represented baseline values. RESULTS: Pairwise agreement among the assessors for any of the three options ranged from 78-89%. At least three experts agreed for 147 cycles (96%), accounting for 28 conception losses and 19 continuing conceptions. The multi-rater kappa was 0.62 for the conception lost category and 0.68 for continuing conceptions, indicating substantial agreement. CONCLUSION: The main sources of disagreement involved deciding whether there was sufficient information for assessment, interpreting cycle parameters such as cycle length or bleeding event, and interpreting a distinct HCG rise pattern that does not exceed the baseline value obtained from the sterilized women. PMID- 11925407 TI - Defective endovascular trophoblast invasion in primary antiphospholipid antibody syndrome-associated early pregnancy failure. AB - BACKGROUND: Primary antiphospholipid antibody syndrome (PAPS) is an established cause of recurrent pregnancy loss, traditionally presumed to be due to 'intraplacental thromboses'. This study examines products of conception (POC) from early pregnancy failures to investigate the mechanism of pregnancy loss. METHODS: POC from patients attending a recurrent miscarriage clinic and from terminations of pregnancy for non-medical reasons were examined histologically with particular regard to the presence or absence of vascular or intervillous thromboses and decidual endovascular trophoblast invasion. RESULTS: There were 31 PAPS-positive, 50 PAPS-negative, 34 aneuploid and 20 control cases at 6-14 weeks gestation. Villous morphology and frequency of intervillous thrombosis were not different among groups. Normal intradecidual endovascular trophoblast invasion was identified significantly less frequently in PAPS+ cases (24%), compared with controls (75%), aneuploid (53%), or PAPS- cases (61%; Z = -3.0, P < 0.01). In all cases there was apparently normal interstitial extravillous trophoblast invasion. CONCLUSIONS: Defective decidual endovascular trophoblast invasion, rather than excessive intervillous thrombosis, is the most frequent histological abnormality in PAPS+ associated early pregnancy loss. Furthermore, endovascular trophoblast invasion is not significantly reduced in the majority of fetal aneuploidy associated pregnancy failures. PMID- 11925408 TI - Altered phenotype of HLA-G expressing trophoblast and decidual natural killer cells in pathological pregnancies. AB - BACKGROUND: The interaction between decidual natural killer (NK) cells and alloantigens expressed on fetal trophoblast cells are thought to be essential for successful implantation and placentation. Consequently, a disturbed interaction during the first trimester of pregnancy might well lead to a subsequent pregnancy failure. METHODS: We investigated the expression of HLA-G and NK cell markers in tissue sections from recurrent miscarriage (n = 9) and ectopic tubal pregnancies (n = 5), and two hysterectomy specimens of healthy pregnancy as well as decidual biopsies (n = 9) were used as controls. RESULTS: We show in normal pregnancy not only a decrease, but also a morphological change in CD56+ NK cells upon interaction with HLA-G-expressing trophoblasts. The cells appear to be transitioning from a blast-like (activation) state into a state of apoptosis. The number of CD16+ NK cells was low. In contrast, in recurrent miscarriage tissue a sustained NK cell marker expression of both CD56 and CD16 was paralleled by a decreased expression of HLA-G. No morphological changes from the blast-like stage were apparent. Finally, in ectopic pregnancies HLA-G expression in the absence of decidual NK cells was associated with a disturbed trophoblast differentiation. CONCLUSIONS: In pathological pregnancies we show an in-situ altered phenotype of trophoblast and NK cells. PMID- 11925409 TI - Serial first- and second-trimester Down's syndrome screening tests among IVF versus naturally-conceived singletons. AB - BACKGROUND: It has been reported that second-trimester serum markers may be affected by assisted reproduction, leading to a higher false-positive rate. METHODS: A total of 285 naturally and 71 IVF-conceived singletons which underwent a serial disclosure Down's syndrome screening programme were compared. The study protocol included first-trimester combined [nuchal translucency (NT), free beta HCG and pregnancy-associated plasma protein-A (PAPP-A)] testing. The second trimester triple serum screening included alpha-fetoprotein (AFP), intact HCG and unconjugated estriol (uE3). After excluding aneuploidies, miscarriages, anatomical anomalies and cases with incomplete follow-up, the serum samples of normal cases were assessed and correlated. RESULTS: NT measurement was not significantly changed in either group. However, the IVF group had lower PAPP-A [0.96 versus 1.05 multiples of normal median (MoM)] and higher AFP (1.13 versus 1.07 median MoM). Both groups had similar rates of first-trimester false-positive results (FPR; 7 and 9% respectively), but the IVF group had a significantly higher mid-gestation FPR rate (10 versus 5%; Pearson chi2, P = 0.029). This has contributed to amniocentesis uptake rates of 15 and 13% for the IVF and natural conception pregnancies respectively. CONCLUSIONS: The IVF group tended to have a significantly higher second-trimester FPR rate. To counterbalance this phenomenon, integrated first- and second-trimester screening tests or the use of NT alone might be a reasonable option that deserves further investigation. PMID- 11925410 TI - Increased nuchal translucency is associated with jugular lymphatic distension. AB - BACKGROUND: Measurement of nuchal translucency (NT) is a widely used method of screening for chromosomal abnormalities. Increased NT is seen in a diversity of fetal malformations. The mechanism explaining the abnormal fluid accumulation and the transient nature of NT remains unexplained. METHODS: The nuchal regions of normal and trisomy 16 mouse embryos were examined for (lympho)vascular abnormalities using immunohistochemical markers against lymphatic vessels (LYVE 1) and smooth muscle (1A4) and endothelial (CD34) cells. Additionally, an ultrasonographic study was carried out on 17 human fetuses with an increased NT. Two of these fetuses were examined morphologically. RESULTS: In both abnormal human and mouse specimens, we found a mesenchyme lined cavity within the posterior nuchal region as well as bilaterally enlarged jugular LYVE-1 positive lymphatic sacs. The persistence of jugular lymphatic sacs was also confirmed by ultrasound in 14 human fetuses with increased NT. CONCLUSION: Our findings identify the cause of increased NT as mesenchymal oedema in the presence of distended jugular lymphatic sacs, detected by the hyaluronan receptor LYVE-1. The delayed organization and connection of these lymphatic sacs to the venous circulation might explain the transient nature of NT. Disturbance in timing of endothelial differentiation might be a common denominator in the origin of NT, linking cardiovascular and haemodynamic abnormalities. PMID- 11925411 TI - Down's syndrome screening with nuchal translucency at 12(+0)-14(+0) weeks and maternal serum markers at 14(+1)-17(+0) weeks: a prospective study. AB - BACKGROUND: Sonographic and biochemical methods for Down's syndrome screening have developed simultaneously, but independently. As a consequence, the rate of invasive procedures for fetal karyotyping has dramatically increased and become an important public health issue which needs to be controlled. One approach is to combine sonographic and biochemical results into a single risk assessment. METHODS: In a multicentre interventional study, nuchal translucency (NT) was measured between 12(+0) and 14(+0) weeks of gestation. Maternal serum markers (MSM) were measured between 14(+1) and 17(+0) weeks of gestation. Karyotyping was advised when: (i) NT was > or =3 mm; or (ii) the MSM-related risk was > or =1 in 250 at term. Karyotyping was delayed until after a maternal blood sample had been taken. NT and MSM were expressed as multiples of the medians (MoMs), and risks were calculated and tailored to the study population. A combined risk for NT and MSM was estimated retrospectively. Costs per case diagnosed, and the cost per case averted were calculated for the three screening strategies. RESULTS: A total of 9444 women was screened. Twenty-one fetuses (0.22%) had Down's syndrome, whilst 326 women (3.4%) were lost to follow-up. Among 9118 women followed up, 5506 had both NT and MSM, 821 had only NT, and 2791 had only MSM. Median maternal age was 30.5 years. False-positive rates for NT, MSM and NT combined with MSM were 3.0, 5.8 and 0.23% respectively. The false-positive rate generated by a sequential two-stage screening was 8.6%. Detection rates of Down's syndrome were 62 and 55% for NT and MSM respectively. Seven cases with Down's syndrome (35%) had raised NT and MSM, and 17 (81%) had either raised NT, MSM, or both. For a 5% false-positive rate, detection rates were 55 and 80% for NT alone and for combined NT and MSM respectively. Ultrasound alone appears to be more cost effective ( pound50 per case diagnosed) than both tests ( pound61 per case diagnosed). CONCLUSIONS: The study results suggest a 25% increase in the detection rate of Down's syndrome using a combination of NT measurement at 12(+0) 14(+0) weeks and MSM at 14(+1)-17(+0) weeks for a 5% false-positive rate, with modest increase in cost. PMID- 11925412 TI - Spontaneous intrauterine pregnancy following salpingectomy for a unilateral hydrosalpinx. AB - The effect of salpingectomy for unilateral hydrosalpinx with a contralateral normal tube was evaluated in two infertile patients in which unilateral hydrosalpinx, visualized by vaginal ultrasound, was treated by unilateral salpingectomy as a preparatory step before IVF. Spontaneous pregnancy occurred in both patients while waiting to be enrolled in an IVF trial. In conclusion, unilateral salpingectomy for hydrosalpinx in the presence of a contralateral healthy tube could result in spontaneous pregnancy. PMID- 11925413 TI - Association of polymorphisms of the oestrogen receptor alpha gene with the age of menarche. AB - BACKGROUND: The age of menarche may be subject to hereditary influences, but the specific genetic determinants are largely unknown. We evaluated whether the XbaI and PvuII polymorphisms of the estrogen receptor alpha gene are associated with the age of menarche. METHODS: We performed genotyping for XbaI and PvuII in a cohort of 145 adolescent females from a closed community in North-Western Greece. RESULTS: There was strong linkage disequilibrium between the two polymorphisms. Menarche occurred later in girls with the XX genotype than in girls with the Xx or xx genotype (mean +/- SD: 13.36 +/- 1.24 versus 12.80 +/- 1.14 and 12.75 +/- 1.35 years respectively; P = 0.017). Menarche also tended to occur later in PP homozygotes than in Pp and pp subjects, but the difference was not significant (mean +/- SD: 13.09 +/- 1.29 versus 12.80 +/- 1.19 and 12.85 +/- 1.33 years respectively). The strongest effect was seen when the PX haplotype was considered [mean +/- SD: 13.43 +/- 1.18 years for homozygotes versus 12.76 +/- 1.25 years in heterozygotes and in subjects without the PX allele, P = 0.006]. CONCLUSIONS: We document that the XbaI polymorphism, and possibly PvuII, may be genetic determinants of the age of menarche. PMID- 11925414 TI - Is there a link between an extremely poor response to ovarian hyperstimulation and early ovarian failure? AB - BACKGROUND: It has been previously reported that a group of 12 infertile women, who had a normal baseline hormonal profile and did not respond to repeated ovarian stimulation with gonadotrophins, developed ovarian failure within a few months. Based on this observation, we carried out a controlled retrospective cohort study to examine whether non-response to ovarian stimulation is linked to early ovarian failure. METHODS: All patients aged 35-40 years who had cancelled IVF cycles for non-response between 1991 and 1993 in our centre were asked to report on the subsequent development of menopausal symptoms, menopause or commencement of hormone replacement therapy. A control group consisted of patients with the same age and similar medical history, who had IVF the same year and responded well. RESULTS: Eleven out of the 12 patients of the non-response group developed menopausal symptoms within 7 years, compared with only four out of 24 in the control group. Similarly, eight out of 12 non-responders either went into menopause or started using hormone replacement therapy compared with one out of 24 in the control group. Using Fisher's exact test, the differences were highly significant (P < 0.0001). The median age at development of menopausal symptoms in the study group was 40 years (range 38-45). The median time between non-response and development of menopausal symptoms was 4 years (range 1-7). CONCLUSION: We carried out a controlled retrospective cohort study that showed a strong association between an extremely poor response to ovarian hyperstimulation and early ovarian failure. PMID- 11925415 TI - A high hypospadias rate in The Netherlands. AB - BACKGROUND: Reports on increasing hypospadias trends are based on birth defect registries, which are prone to inaccuracy. We assessed the prevalence of hypospadias precisely, by prospective examination of all newborns in Rotterdam over a 2-year period. METHODS: A total of 7292 consecutive male births were examined for the presence of hypospadias, classified by severity. RESULTS: The frequency of hypospadias in newborn boys was 0.73% (53/7292). The rate among live births was 38 per 10 000, which is 6 times the previously reported rate for the Southwestern Netherlands (6.2) (P < 0.0001). This registry excludes glandular hypospadias. Without glandular cases, our rate is 26 per 10 000, which is still 4 fold higher (P < 0.0001). The ratio of minor to major hypospadias was 0.3. In 79% of cases, surgery was indicated. CONCLUSIONS: We found a 4-fold higher than expected hypospadias rate, which may be explained by case ascertainment differences. The proportion of major cases was higher than generally assumed. This study provides evidence for substantial geographical differences. Explanations for temporal and geographical differences need to be explored. To monitor hypospadias rates and trends accurately, complete case ascertainment, including standardized classification of severity, is warranted. PMID- 11925416 TI - Parenting and psychosocial development of IVF children: a follow-up study. AB - BACKGROUND: This report details a follow-up study of the parent-child relationship and the child's psychosocial development after IVF. The pilot study compared 31 IVF families and 31 families with a naturally conceived child when the children were aged 2 years. Twenty-seven IVF and 23 control families participated again when the children were aged 8-9 years. METHODS: Fathers and mothers completed questionnaires assessing parenting variables and the child's behaviour. For most children, behavioural ratings were also obtained from the child's teacher. RESULTS: No significant differences were found between IVF and control parents' reports of child behaviour, parenting behaviour, parenting stress and most of the parenting goals. The parenting goal adjustment was significantly more important for IVF than for control fathers; religion was more important for IVF than for naturally conceiving mothers. Teacher ratings of the child's behaviour did not differ significantly between the IVF and control groups. All couples but one had talked to other persons about the IVF conception; 75% of the IVF parents had not yet informed their children. IVF parents who had informed their child observed more (internalizing and for fathers also overall) problem behaviours in their child, compared with IVF parents who had not yet disclosed the IVF conception. CONCLUSIONS: Parenting and the children's psychosocial development do not differ significantly between IVF families and control families. PMID- 11925417 TI - An acute psychiatric episode following transvaginal oocyte retrieval. AB - Psychological disorders of infertile patients are traditionally thought to be chronic, to advance gradually, and to be long-term problems. We describe a patient in whom an acute psychiatric episode developed immediately after transvaginal ultrasound-guided oocyte retrieval. A 34 year old women without history of psychiatric disturbance or adverse reaction to drugs suffered an acute psychiatric episode immediately after oocyte retrieval. She exhibited tachycardia, tachypnoea, transient hypertension and limb rigidity, as well as alterations to stupor and posture. Her vital signs stabilized and she opened her eyes 6 h later, but she persistently raised her head to the left and stared blankly without response to external stimuli. Nine hours later, she was able to look around but remained unresponsive to stimuli. Aphasia was noted in the next morning and a wishful thinking of having delivered a baby was noted in the afternoon. Memory loss was noted on the third day. The patient was diagnosed as having dissociative amnesia and was discharged after three courses of supportive psychotherapy. Assisted reproductive technology-related acute psychiatric episodes, which may initially mimic brainstem stroke, are rare; however, attention should be paid to high-risk patients, and they should be offered elective psychological counselling. PMID- 11925418 TI - Testicular sperm extraction in azoospermic cancer patients prior to treatment--a new guideline? PMID- 11925419 TI - Pelvic congestion/chronic pain dynamics. PMID- 11925420 TI - Treatment of chronic pelvic pain in patients with endometriosis. PMID- 11925421 TI - How homogeneous are comparison groups in any study evaluating techniques of embryo transfer? PMID- 11925424 TI - Ultrasound-guided embryo transfer maximizes the IVF results on day 3 and day 4 embryo transfer but has no impact on day 5. PMID- 11925426 TI - Substrate recognition drives the evolution of serine proteases. AB - A method is introduced to identify amino acid residues that dictate the functional diversity acquired during evolution in a protein family. Using over 80 enzymes of the chymotrypsin family, we demonstrate that the general organization of the phylogenetic tree and its functional branch points are fully accounted for by a limited number of residues that cluster around the active site of the protein and define the contact region with the P1-P4 residues of substrate. PMID- 11925427 TI - The connecting segment between both epidermal growth factor-like domains in blood coagulation factor IX contributes to stimulation by factor VIIIa and its isolated A2 domain. AB - The light chain of activated factor IX comprises multiple interactions between both epidermal growth factor-like domains that contribute to enzymatic activity and binding of factor IXa to its cofactor factor VIIIa. To investigate the association between factor IXa-specific properties and surface-exposed structure elements, chimeras were constructed in which the interconnection between the modules Leu(84)-Thr(87) and the factor IX-specific loop Asn(89)-Lys(91) were exchanged for corresponding regions of factor X and factor VII. In absence of factor VIIIa, all chimeras displayed normal enzymatic activity. In the presence of factor VIIIa, replacement of loop Asn(89)-Lys(91) resulted in a minor reduction in factor IXa activity. However, chimeras with substitutions or insertions in the spacer between the epidermal growth factor-like domains showed a major defect in response to factor VIIIa. Of these chimeras, some displayed a normal response to isolated factor VIII A2 domain as a cofactor in factor X activation. Surprisingly, chimeras containing elongated inter-domain spacers from factor X or VII displayed reduced response to both complete factor VIIIa and the isolated A2 domain. Moreover, these chimeras still displayed effective association with immobilized A2 domain as assessed by surface plasmon resonance. We conclude that both sequence and length of the junction Leu(84)-Thr(87) between both epidermal growth factor-like domains contribute to the enhancement of factor IXa enzymatic activity that occurs upon assembly with factor VIIIa. PMID- 11925429 TI - Direct, Ca2+-dependent interaction between tubulin and synaptotagmin I: a possible mechanism for attaching synaptic vesicles to microtubules. AB - The synaptic vesicle protein synaptotagmin I probably plays important roles in the synaptic vesicle cycle. However, the mechanisms of its action remain unclear. In this study, we have searched for cytoplasmic proteins that interact with synaptotagmin I. We found that the cytoskeletal protein tubulin directly and stoichiometrically bound to recombinant synaptotagmin I. The binding depended on mm Ca(2+), and 1 mol of tubulin dimer bound 2 mol of synaptotagmin I with half maximal binding at 6.6 microm tubulin. The Ca(2+) dependence mainly resulted from Ca(2+) binding to the Ca(2+) ligands of synaptotagmin I. The C-terminal region of beta-tubulin and both C2 domains of synaptotagmin I were involved in the binding. The YVK motif in the C2 domains of synaptotagmin I was essential for tubulin binding. Tubulin and synaptotagmin I were co-precipitated from the synaptosome extract with monoclonal antibodies to tubulin and SNAP-25 (synaptosome-associated protein of 25 kDa), indicating the presence of tubulin/synaptotagmin I complex and tubulin binding to synaptotagmin I in SNARE (soluble N-ethylmaleimide sensitive factor attachment protein receptor) complexes. Synaptotagmin I promoted tubulin polymerization and bundled microtubules in the presence of Ca(2+). These results suggest that direct interaction between synaptotagmin I and tubulin provides a mechanism for attaching synaptic vesicles to microtubules in high Ca(2+) concentrations. PMID- 11925428 TI - Influence of peroxisome proliferator-activated receptor alpha agonists on the intracellular turnover and secretion of apolipoprotein (Apo) B-100 and ApoB-48. AB - The peroxisome proliferator-activated receptor (PPAR) alpha agonist WY 14,643 increased the secretion of apolipoprotein (apo) B-100, but not that of apoB-48, and decreased triglyceride biosynthesis and secretion from primary rat hepatocytes. These effects resulted in decreased secretion of apoB-100-very low density lipoprotein (VLDL) and an increased secretion of apoB-100 on low density lipoproteins/intermediate density lipoproteins. ApoB-48-VLDL was also replaced by more dense particles. The proteasomal inhibitor lactacystin did not influence the recovery of apoB-100 or apoB-48 in primary rat hepatocytes, indicating that co translational (proteasomal) degradation is of less importance in these cells. Treatment with WY 14,643 made the recovery of apoB-100 sensitive to lactacystin, most likely reflecting the decreased biosynthesis of triglycerides. The PPAR alpha agonist induced a significant increase in the accumulation of pulse-labeled apoB-100 even after a short pulse (2-5 min). There was also an increase in apoB 100 nascent polypeptides, indicating that the co-translational degradation of apoB-100 was inhibited. However, a minor influence on an early posttranslation degradation cannot be excluded. This decreased co-translational degradation of apoB-100 explained the increased secretion of the protein. The levels of apoB-48 remained unchanged during these pulse-chase experiments, and albumin production was not affected, indicating a specific effect of PPAR alpha agonists on the co translational degradation of apoB-100. These findings explain the difference in the rate of secretion of the two apoB proteins seen after PPAR alpha activation. PPAR alpha agonists increased the expression and biosynthesis of liver fatty acid binding protein (LFABP). Increased expression of LFABP by transfection of McA RH7777 cells increased the secretion of apoB-100, decreased triglyceride biosynthesis and secretion, and increased PPAR alpha mRNA levels. These findings suggest that PPAR alpha and LFABP could interact to amplify the effect of endogenous PPAR alpha agonists on the assembly of VLDL. PMID- 11925430 TI - Identification and characterization of HIPK2 interacting with p73 and modulating functions of the p53 family in vivo. AB - To study the biological role of p73 alpha, a member of the p53 tumor suppressor family, we performed a yeast two-hybrid screen of a human cDNA library. Using a p73 alpha fragment consisting of amino acids 49-636 as bait, we found that p73 alpha is functionally associated with the human homologue of mouse and hamster homeodomain-interacting protein kinase 2 (HIPK2). The hamster homologue, also known as haHIPK2 or PKM, was used for further characterization of interactions between HIPK2 and members of the p53 protein family. Systematic yeast two-hybrid assays indicated a physical interaction between the oligomerization domains of p73 alpha and p53 (amino acid regions 345-380 and 319-360, respectively) and amino acid region 812-907 of haHIPK2. This region of haHIPK2 includes a PEST sequence, an Ubc9-binding domain, and a partial speckle retention sequence and is identical to amino acid residues 846-941 of human HIPK2 (hHIPK2). The interaction was confirmed by glutathione S-transferase pull-down assays in vitro and immunoprecipitation assays in vivo. HIPK2 colocalized with p73 and p53 in nuclear bodies, as shown by confocal microscopy. Overexpression of HIPK2 stabilized the p53 protein and greatly increased the p73- and p53-induced transcriptional repression of multidrug-resistant and collagenase promoters in Saos2 cells but had little effect on the p73- or p53-mediated transcriptional activation of synthetic p53-responsive and p21WAF1 promoters. Stable expression of HIPK2 in U2OS cells enhanced the cisplatin response of sub-G(1) and G(2)/M populations, and it also increased the apoptotic response to cisplatin and adriamycin as demonstrated by fluorescence-activated cell sorter and 4',6-diamidino-2 phenylindole-staining analyses. HIPK2 potentiated the inhibition of colony formation by p73 and p53. These results suggest that physical interactions between HIPK2 and members of the p53 family may determine the roles of these proteins in cell cycle regulation and apoptosis. PMID- 11925431 TI - Distinct modes of cell death induced by different reactive oxygen species: amino acyl chloramines mediate hypochlorous acid-induced apoptosis. AB - Oxidants derived from inflammatory phagocytes compose a key element of the host immune defense system and can kill mammalian cells by one of several different mechanisms. In this report, we compare mechanisms of cell death induced in human B lymphoma cells by the inflammatory oxidants superoxide, H(2)O(2), and HOCl. The results indicate that the mode of cell death induced depends on the nature of the oxidant involved and the medium in which the cells are treated. When human Burkitt's lymphoma cells are exposed to superoxide anion, generated as a flux from xanthine and xanthine oxidase, the cells die by a non-apoptotic mechanism (pyknosis/necrosis) identical to that seen when cells are treated with a bolus of reagent H(2)O(2). Addition of superoxide dismutase has no effect, whereas catalase is completely protective, indicating that exogenously generated superoxide kills cells entirely through its dismutation into H(2)O(2). In contrast, cells treated in culture media with reagent HOCl die largely by apoptosis. HOCl-induced apoptosis is mediated by aminoacyl chloramines generated in the culture media and can be mimicked by treatment of cells with taurine chloramine or with long lived chloramines generated from modified Lys or Arg. The results suggest that in a physiological milieu in which O(2)(-) and H(2)O(2) are the main oxidants being formed, the principal form of cell death may be necrotic, and under inflammatory conditions in which HOCl is generated, apoptotic cell death may predominate. PMID- 11925432 TI - A novel family of calmodulin-binding transcription activators in multicellular organisms. AB - Screening of cDNA expression libraries derived from plants exposed to stress, with 35S-labeled recombinant calmodulin as a probe, revealed a new family of proteins containing a transcription activation domain and two types of DNA binding domains designated the CG-1 domain and the transcription factor immunoglobulin domain, ankyrin repeats, and a varying number of IQ calmodulin binding motifs. Based on domain organization and amino acid sequence comparisons, similar proteins, with the same domain organization, were identified in the genomes of other multicellular organisms including human, Drosophila, and Caenorhabditis, whereas none were found in the complete genomes of single cell eukaryotes and prokaryotes. This family of proteins was designated calmodulin binding transcription activators (CAMTAs). Arabidopsis thaliana contains six CAMTA genes (AtCAMTA1-AtCAMTA6). The transcription activation domain of AtCAMTA1 was mapped by testing a series of protein fusions with the DNA-binding domain of the bacterial LexA transcription factor and two reporter genes fused to LexA recognition sequences in yeast cells. Two human proteins designated HsCAMTA1 and HsCAMTA2 were also shown to activate transcription in yeast using the same reporter system. Subcellular fractionation of Arabidopsis tissues revealed the presence of CAMTAs predominantly in the nucleus. Calmodulin binding assays identified a region of 25 amino acids capable of binding calmodulin with high affinity (K(d) = 1.2 nm) in the presence of calcium. We suggest that CAMTAs comprise a conserved family of transcription factors in a wide range of multicellular eukaryotes, which possibly respond to calcium signaling by direct binding of calmodulin. PMID- 11925433 TI - Secreted and transmembrane mucins inhibit gene transfer with AAV4 more efficiently than AAV5. AB - Adeno-associated virus (AAV) is a promising vector for gene transfer in cystic fibrosis. AAV4 and AAV5 both bind to the apical surface of differentiated human airway epithelia, but only AAV5 infects. Both AAV4 and AAV5 require 2,3-linked sialic acid for binding. However, AAV5 interacts with sialic acid on N-linked carbohydrates, whereas AAV4 interacts with sialic acid on O-linked carbohydrates. Because mucin is decorated with O-linked carbohydrates, we hypothesized that mucin binds AAV4 and inhibits gene transfer. To evaluate the effect of secreted mucin, we studied mucin binding and gene transfer to COS cells and the basolateral membrane of well differentiated human airway epithelia. AAV4 bound mucin more efficiently than AAV5, and mucin inhibited gene transfer with AAV4. Moreover, O-glycosidase-pretreated mucin did not block gene transfer with AAV4. Similar to secreted mucin, the transmembrane mucin MUC1 inhibited gene transfer with AAV4 but not AAV5. MUC1 inhibited AAV4 by blocking internalization of the virus. Thus, O-linked carbohydrates of mucin are potent inhibitors of AAV4. Furthermore, whereas mucin plays an important role in innate host defense, its activity is specific; some vectors or pathogens are more resistant to its effects. PMID- 11925434 TI - Calcium/calmodulin-dependent protein kinase IIdelta 2 and gamma isoforms regulate potassium currents of rat brain capillary endothelial cells under hypoxic conditions. AB - Endothelial K+ and Ca2+ homeostasis plays an important role in the regulation of tissue supply and metabolism under normal and pathological conditions. However, the exact molecular mechanism of how Ca2+ is involved in the regulation of K+ homeostasis in capillary endothelial cells, especially under oxidative stress, is not clear. To reveal Ca2+-triggered pathways, which modulate K+ homeostasis, Ca2+/calmodulin-dependent protein kinase II and voltage-gated outward K+ currents were studied in rat brain capillary endothelial cells under hypoxia. Whole cell voltage-clamp measurements showed voltage-gated outward K+ current with transient and sustained components. mRNA and protein of Ca2+/calmodulin-dependent protein kinase II delta2 and two gamma isoenzymes were identified. Activation of the isoforms (autophosphorylation) was typically achieved by the Ca2+ ionophore ionomycin, which was prevented by the Ca2+/calmodulin-dependent protein kinase II specific inhibitor KN-93. Hypoxia resulted in autophosphorylation of the delta2 and gammaB isoforms, augmented the current amplitude, increased the inactivation time constant, and decreased the extent of inactivation of the transient current. KN-93 prevented both the activation of the isoforms and the alterations in the K+ current characteristics. It is concluded that the activation of Ca2+/calmodulin dependent protein kinase II decreases inactivation of the voltage-gated outward K+ current, thereby counteracting depolarization of the hypoxic endothelium. PMID- 11925435 TI - S-thiolation of HSP27 regulates its multimeric aggregate size independently of phosphorylation. AB - HSP27 exists as large aggregates that breakdown after phosphorylation. We show rat cardiac HSP27 is S-thiolated during oxidant stress, and this modification, without phosphorylation, disaggregates multimeric HSP27. Biotinylated cysteine acts as a probe for thiolated proteins, which are detected using non-reducing Western blots probed with streptavidin-horseradish peroxidase. Controls show a low level of S-thiolation, which is increased 3.6-fold during post-ischemic reperfusion. S-thiolated proteins were purified using streptavidin-agarose, and Western immunoblotting showed HSP27 was present. We increased protein S thiolation 10-fold with 10 microm H2O2 with or without a kinase inhibitor mixture (staurosporine, genistein, bisindolylmaleimide, SB203580, and PD98059). H2O2 alone induced the phosphorylation of HSP27 Ser-86 and Ser-45/Ser-59 of its homologue alphaB crystallin. However, kinase inhibition reduced phosphorylation of these sites below basal. Despite effective kinase inhibition, H2O2 still disaggregated HSP27, but not alphaB crystallin. This is consistent with the lack of an S-thiolation site on alphaB crystallin. Thus, we have demonstrated a novel mechanism of HSP27 multimeric size regulation. S-thiolation must occur at Cys 141, the only cysteine in rat HSP27. PMID- 11925436 TI - BARD1 induces BRCA1 intranuclear foci formation by increasing RING-dependent BRCA1 nuclear import and inhibiting BRCA1 nuclear export. AB - BRCA1 is a tumor suppressor with several important nuclear functions. BRCA1 has no known cytoplasmic functions. We show here that the two previously identified nuclear localization signals (NLSs) are insufficient for nuclear localization of BRCA1 due to the opposing action of an NH2-terminal nuclear export signal. In transfected breast cancer cells, BRCA1 nuclear localization requires both the NLSs and NH2-terminal RING domain region; mutating either of these sequences shifts BRCA1 to the cytoplasm. The BRCA1 RING element mediates nuclear import via association with BARD1, and this is not affected by cancer-associated RING mutations. Moreover, BARD1 directly masks the BRCA1 nuclear export signal, and the resulting block to nuclear export is requisite for efficient import and nuclear localization of ectopic and endogenous BRCA1. Our results explain why BRCA1 exon 11 splice variants, which lack the NLSs but retain the RING domain, are frequently detected in the nucleus and in nuclear foci in vivo. In fact, co expression of BARD1 promoted formation of DNA damage-induced nuclear foci comprising ectopic wild-type or NLS-deficient BRCA1, implicating BARD1 in nuclear targeting of BRCA1 for DNA repair. Our identification of BARD1 as a BRCA1 nuclear chaperone has regulatory implications for its reported effects on BRCA1 protein stability, ubiquitin ligase activity, and DNA repair. PMID- 11925437 TI - Role of the proline-rich domain of dynamin-2 and its interactions with Src homology 3 domains during endocytosis of the AT1 angiotensin receptor. AB - In nonneural tissues, the dynamin-2 isoform participates in the formation of clathrin-coated vesicles during receptor endocytosis. In this study, the mechanism of dynamin-2 action was explored during endocytosis of the G protein coupled AT1A angiotensin receptor expressed in Chinese hamster ovary cells. Dynamin-2 molecules with mutant pleckstrin homology domains or deleted proline rich domains (PRD) exerted dominant negative inhibition on the endocytosis of radiolabeled angiotensin II. However, only the PRD mutation interfered with the localization of the dynamin-2 molecule to clathrin-coated pits and reduced the inhibitory effect of the GTPase-deficient K44A mutant dynamin-2. Green fluorescent protein-tagged Src homology 3 (SH3) domains of endophilin I and amphiphysin II, two major binding partners of dynamins, also inhibited AT1A receptor-mediated endocytosis of angiotensin II. These effects were partially or fully, respectively, restored by the overexpression of dynamin-2. Transient overexpression of these SH3 domains also reduced the localization of dynamin-2 to clathrin-coated pits. These data indicate that, similar to the recruitment of dynamin-1 during the recycling of synaptic vesicles, interaction of the dynamin-2 PRD with SH3 domains of proteins such as the amphiphysins and endophilins is essential for AT1A receptor endocytosis. This mechanism could be of general importance in dynamin-dependent endocytosis of other G protein-coupled receptors in nonneural tissues. PMID- 11925438 TI - Protein kinase C induces actin reorganization via a Src- and Rho-dependent pathway. AB - We have investigated the mechanism of PKC-induced actin reorganization in A7r5 vascular smooth muscle cells. PKC activation by 12-O-tetradecanoylphorbol-13 acetate induces the disassembly of actin stress fibers concomitant with the appearance of membrane ruffles. PKC also induces rapid tyrosine phosphorylation in these cells. As we could show, utilizing the Src-specific inhibitor PP2 and a kinase-deficient c-Src mutant, actin reorganization is dependent on PKC-induced Src activation. Subsequently, the activity of the small G-protein RhoA is decreased, whereas Rac and Cdc42 activities remain unchanged. Disassembly of actin stress fibers could also be observed using the Rho kinase-specific inhibitor Y-27632, indicating that the decrease in RhoA activity on its own is responsible for actin reorganization. In addition, we show that tyrosine phosphorylation of p190RhoGAP is increased upon 12-O-tetradecanoylphorbol-13 acetate stimulation, directly linking Src activation to a decrease in RhoA activity. Our data provide substantial evidence for a model elucidating the molecular mechanisms of PKC-induced actin rearrangements. PMID- 11925440 TI - Mapping of the catalytic groove preferences of factor Xa reveals an inadequate selectivity for its macromolecule substrates. AB - Factor Xa (FXa) hydrolyzes two peptide bonds in prothrombin having (Glu/Asp)-Gly Arg-(Thr/Ile) for P(3)-P(2)-P(1)-P(1)' residues, but the exact preferences of its catalytic groove remain largely unknown. To investigate the specificity of FXa, we synthesized full sets of fluorescence-quenched substrates carrying all natural amino acids (except Cys) in P(3), P(2), P(1)', P(2)', and P(3)' and determined the k(cat)/K(m) values of cleavage. Contrary to expectation, glycine was not the "best" P(2) residue; peptide with phenylalanine was cleaved slightly faster. In fact, FXa had surprisingly limited preferences, barely more pronounced than trypsin; in P(2), the ratio of the k(cat)/K(m) values for the most favorable side chain over the least was 289 (12 with trypsin), but in P(1)', this ratio was only 30 (versus 80 with trypsin). This unexpected selectivity undoubtedly distinguished FXa from thrombin, which exhibited ratios higher than 19,000 in P(2) and P(1)'. Thus, with respect to the catalytic groove, FXa resembles a low efficiency trypsin rather than the highly selective thrombin. The rates of cleavage of the peptidyl substrates were virtually identical whether or not FXa was in complex with factor Va, suggesting that the cofactor did not exert a direct allosteric control on the catalytic groove. We conclude that the remarkable efficacy of FXa within prothrombinase originates from exosite interaction(s) with factor Va and/or prothrombin rather than from the selectivity of its catalytic groove. PMID- 11925439 TI - The 25-kDa synaptosome-associated protein (SNAP-25) binds and inhibits delayed rectifier potassium channels in secretory cells. AB - Delayed-rectifier K(+) channels (K(DR)) are important regulators of membrane excitability in neurons and neuroendocrine cells. Opening of these voltage dependent K(+) channels results in membrane repolarization, leading to the closure of the Ca(2+) channels and cessation of insulin secretion in neuroendocrine islet beta cells. Using patch clamp techniques, we have demonstrated that the activity of the K(DR) channel subtype, K(V)1.1, identified by its specific blocker dendrodotoxin-K, is inhibited by SNAP-25 in insulinoma HIT-T15 beta cells. A co-precipitation study of rat brain confirmed that SNAP-25 interacts with the K(V)1.1 protein. Cleavage of SNAP-25 by expression of botulinum neurotoxin A in HIT-T15 cells relieved this SNAP-25-mediated inhibition of K(DR). This inhibitory effect of SNAP-25 is mediated by the N terminus of K(V)1.1, likely by direct interactions with K(Valpha)1.1 and/or K(V)beta subunits, as revealed by co-immunoprecipitation performed in the Xenopus oocyte expression system and in vitro binding. Taken together we have concluded that SNAP-25 mediates secretion not only through its participation in the exocytotic SNARE complex but also by regulating membrane potential and calcium entry through its interaction with K(DR) channels. PMID- 11925441 TI - Determinants of the substrate specificity of multidrug resistance protein 1: role of amino acid residues with hydrogen bonding potential in predicted transmembrane helix 17. AB - Human multidrug resistance protein 1 (MRP1) confers resistance to many natural product chemotherapeutic agents and actively transports structurally diverse organic anion conjugates. We previously demonstrated that two hydrogen-bonding amino acid residues in the predicted transmembrane 17 (TM17) of MRP1, Thr(1242) and Trp(1246), were important for drug resistance and 17beta-estradiol 17-(beta-d glucuronide) (E(2)17betaG) transport. To determine whether other residues with hydrogen bonding potential within TM17 influence substrate specificity, we replaced Ser(1233), Ser(1235), Ser(1237), Gln(1239), Thr(1241), and Asn(1245) with Ala and Tyr(1236) and Tyr(1243) with Phe. Mutations S1233A, S1235A, S1237A, and Q1239A had no effect on any substrate tested. In contrast, mutations Y1236F and T1241A decreased resistance to vincristine but not to VP-16, doxorubicin, and epirubicin. Mutation Y1243F reduced resistance to all drugs tested by 2-3-fold. Replacement of Asn(1245) with Ala also decreased resistance to VP-16, doxorubicin, and epirubicin but increased resistance to vincristine. This mutation also decreased E(2)17betaG transport approximately 5-fold. Only mutation Y1243F altered the ability of MRP1 to transport both leukotriene 4 and E(2)17betaG. Together with our previous results, these findings suggest that residues with side chain hydrogen bonding potential, clustered in the cytoplasmic half of TM17, participate in the formation of a substrate binding site. PMID- 11925442 TI - Mitosis-specific activation of LIM motif-containing protein kinase and roles of cofilin phosphorylation and dephosphorylation in mitosis. AB - Actin filament dynamics play a critical role in mitosis and cytokinesis. LIM motif-containing protein kinase 1 (LIMK1) regulates actin reorganization by phosphorylating and inactivating cofilin, an actin-depolymerizing and -severing protein. To examine the role of LIMK1 and cofilin during the cell cycle, we measured cell cycle-associated changes in the kinase activity of LIMK1 and in the level of cofilin phosphorylation. Using synchronized HeLa cells, we found that LIMK1 became hyperphosphorylated and activated in prometaphase and metaphase, then gradually returned to the basal level as cells entered into telophase and cytokinesis. Although Rho-associated kinase and p21-activated protein kinase phosphorylate and activate LIMK1, they are not likely to be involved in mitosis specific activation and phosphorylation of LIMK1. Immunoblot and immunofluorescence analyses using an anti-phosphocofilin-specific antibody revealed that the level of cofilin phosphorylation, similar to levels of LIMK1 activity, increased during prometaphase and metaphase then gradually declined in telophase and cytokinesis. Ectopic expression of LIMK1 increased the level of cofilin phosphorylation throughout the cell cycle and induced the formation of multinucleate cells. These results suggest that LIMK1 is involved principally in control of mitosis-specific cofilin phosphorylation and that dephosphorylation and reactivation of cofilin at later stages of mitosis play a critical role in cytokinesis of mammalian cells. PMID- 11925443 TI - Redox regulation of Cdc25C. AB - The Cdc25 family of dual specific phosphatases are critical components of cell cycle progression and checkpoint control. Certain stresses such as ultraviolet light stimulate the rapid and selective destruction of Cdc25A protein through a Chk1 protein kinase-dependent pathway. We demonstrate that in contrast to cellular stresses previously examined, hydrogen peroxide exposure affects Cdc25C but not Cdc25A levels. Pharmacological inhibition of Chk1 activity or a mutant of Cdc25C that lacks the Chk1 phosphorylation site still undergoes degradation in response to oxidants. We also demonstrate that in vitro hydrogen peroxide stimulates an intramolecular disulfide bond between the active site cysteine at position 377 and another invariant cysteine at position 330. The in vivo stability of Cdc25C is substantially reduced by the mutation of either of these two cysteine residues. In contrast, a double (C2) mutant of both cysteine 330 and cysteine 377 results in a protein that is more stable than wild type Cdc25C and is resistant to oxidative stress-induced degradation. In addition, the C2 mutant, which is unable to form an intramolecular disulfide bond, has reduced binding to 14-3-3 in vitro and in vivo. These results suggest that oxidative stress may induce cell cycle arrest in part through the degradation of Cdc25C. PMID- 11925444 TI - ATF-1 mediates protease-activated receptor-1 but not receptor tyrosine kinase induced DNA synthesis in vascular smooth muscle cells. AB - Previously we have demonstrated that activation of p38 mitogen-activated protein kinase (MAPK) and induction of DNA synthesis in response to receptor tyrosine kinase (RTK) and G protein-coupled receptor (GPCR) agonists require NADH/NADPH like oxidase activity in vascular smooth muscle cells (VSMC). Here we tested the role of p38 MAPK in RTK and GPCR agonist-induced DNA synthesis in VSMC. Platelet derived growth factor (PDGF)-BB and thrombin (RTK and GPCR agonists, respectively) activated p38 MAPK in a time-dependent manner in VSMC. Inhibition of p38 MAPK led to a 50% decrease in the DNA synthesis induced by thrombin but not PDGF-BB. ATF-1 was found to be the predominant member of the cyclic AMP response element (CRE)-DNA complex formed in VSMC in response to PDGF-BB and thrombin, and both agonists induced its phosphorylation. Regardless of this, inhibition of p38 MAPK reduced only thrombin- but not PDGF-BB-induced ATF-1 phosphorylation. Similarly, inhibition of p38 MAPK caused a 50% decrease in thrombin- but not PDGF-BB-induced CRE promoter-dependent transcription. Ectopic expression of an inhibitory anti-ATF-1 single-chain antibody fragment, ScFv, significantly interfered with DNA synthesis induced by thrombin but not PDGF-BB. Together, these results suggest the following conclusions. 1) Both RTK and GPCR agonists activate p38 MAPK and induce CRE promoter-dependent transcription; 2) both RTK and GPCR agonists induce ATF-1 phosphorylation, and ATF-1 is a predominant member in the CRE-DNA complexes formed in response to these agents; and 3) p38 MAPK-dependent ATF-1 phosphorylation and CRE promoter-mediated transcription are associated with GPCR agonist-induced VSMC growth. PMID- 11925445 TI - Horseradish peroxidase-catalyzed oligomerization of ferulic acid on a template of a tyrosine-containing tripeptide. AB - Ferulic acid (FA) is an abundantly present phenolic constituent of plant cell walls. Kinetically controlled incubation of FA and the tripeptide Gly-Tyr-Gly (GYG) with horseradish peroxidase and H2O2 yielded a range of new cross-linked products. Two predominant series of hetero-oligomers of FA linked by dehydrogenation to the peptidyl tyrosine were characterized by electrospray ionization (tandem) mass spectrometry. One series comprises GYG coupled with 4-7 FA moieties linked by dehydrogenation, of which one is decarboxylated. In the second series 4-9 FA moieties linked by dehydrogenation, of which two are decarboxylated, are coupled to the tripeptide. A third series comprises three hetero-oligomers in which the peptidyl tyrosine is linked to 1-3 FA moieties of which none is decarboxylated. Two mechanisms for the formation of the FA-Tyr oligomers that result from the dualistic, concentration-dependent chemistry of FA and their possible role in the regulation of plant cell wall tissue growth are presented. PMID- 11925446 TI - A UDP-GalNAc:polypeptide N-acetylgalactosaminyltransferase is essential for viability in Drosophila melanogaster. AB - We report the first demonstration that the activity of a member of the UDP GalNAc:polypeptide N-acetylgalactosaminyltransferase gene family is necessary for viability in Drosophila melanogaster. Expression of the wild-type recombinant pgant35A gene in COS7 cells resulted in in vitro activity against peptide and glycopeptide substrates, demonstrating that this gene encodes a biochemically active transferase. Previous mutagenesis studies identified recessive lethal mutations that were rescued by a genomic fragment containing the pgant35A gene; however, the presence of additional open reading frames within this fragment left open the possibility that another gene was responsible for rescue of the observed lethality. Here, we have determined the molecular nature of the mutations in three independent mutant alleles. Two of the mutant alleles contain premature stop codons within the coding region of pgant35A. The third mutant contains an arginine to tryptophan amino acid change, which, when expressed in COS7 cells, resulted in a dramatic reduction of transferase activity in vitro. PCR amplification of this gene from Drosophila cDNA panels and Northern analysis revealed that it is expressed throughout embryonic, larval, and pupal stages as well as in adult males and females. This study provides the first direct evidence for the involvement of a member of this conserved multigene family in eukaryotic development and viability. PMID- 11925447 TI - Chemical quenched flow kinetic studies indicate an intraholoenzyme autophosphorylation mechanism for Ca2+/calmodulin-dependent protein kinase II. AB - Autophosphorylation of alpha-Ca(2+)/calmodulin-dependent protein kinase II (CaM kinase II) at Thr-286 generates Ca(2+)-independent activity that outlasts the initial Ca(2+) stimulus. Previous studies suggested that this autophosphorylation occurs between subunits within each CaM kinase II holoenzyme. However, electron microscopy studies have questioned this mechanism because a large distance separates a kinase domain from its neighboring subunit. Moreover, the recently discovered ability of CaM kinase II holoenzymes to self-associate has raised questions about data interpretation in previous investigations of autophosphorylation. In this work, we characterize the mechanism of CaM kinase II autophosphorylation. To eliminate ambiguity arising from kinase aggregation, we used dynamic light scattering to establish the monodispersity of all enzyme solutions. We then found using chemical quenched flow kinetics that the autophosphorylation rate was independent of the CaM kinase II concentration, results corroborating intraholoenzyme activation. Experiments with a monomeric CaM kinase II showed that phosphorylation of this construct is intermolecular, supporting intersubunit phosphorylation within a holoenzyme. The autophosphorylation rate at 30 degrees C was approximately 12 s(-1), more than 10 fold faster than past estimates. The ability of CaM kinase II to autophosphorylate through an intraholoenzyme, intersubunit mechanism is likely central to its functions of decoding Ca(2+) spike frequency and providing a sustained response to Ca(2+) signals. PMID- 11925448 TI - Notch3 signaling in vascular smooth muscle cells induces c-FLIP expression via ERK/MAPK activation. Resistance to Fas ligand-induced apoptosis. AB - Mutations in the Notch3 receptor result in the cerebral autosomal dominant arteriopathy with subcortical infarcts and leukoencephelopathy (CADASIL) syndrome, a heritable arteriopathy predisposing to early onset stroke. Based upon clinical evidence that CADASIL arteriopathy results in degeneration and loss of vascular smooth muscle cells (VSMC) from the arterial wall, we postulated that Notch3 signaling is a critical determinant of VSMC survival. We initially established that both transient and constitutive Notch3 signaling promoted VSMC survival in response to the proapoptotic Fas ligand (FasL). Resistance to FasL induced apoptosis was associated with the induction of c-FLIP, a primary inhibitor of the FasL signaling pathway. We determined that Notch3's regulation of c-FLIP was independent of the activity of the classical DNA-binding protein, RBP-Jk, but dependent upon cross-talk activation of the ERK/MAPK pathway. We extended our observations to the in vivo context by determining a coordinate regulation of Notch3 and c-FLIP within the arterial wall in response to injury. Furthermore, we defined that expression levels of Notch3 and c-FLIP are coordinately up-regulated within the neointima of remodeled arteries. Taken together, these findings provide initial evidence that Notch3 signaling may be a critical determinant of VSMC survival and vascular structure by modulating the expression of downstream mediators of apoptosis via signaling cross-talk with the ERK/MAPK pathway. PMID- 11925449 TI - The conformationally flexible S9-S10 linker region in the core domain of p53 contains a novel MDM2 binding site whose mutation increases ubiquitination of p53 in vivo. AB - Although the N-terminal BOX-I domain of the tumor suppressor protein p53 contains the primary docking site for MDM2, previous studies demonstrated that RNA stabilizes the MDM2.p53 complex using a p53 mutant lacking the BOX-I motif. In vitro assays measuring the specific activity of MDM2 in the ligand-free and RNA bound state identified a novel MDM2 interaction site in the core domain of p53. As defined using phage-peptide display, the RNA.MDM2 isoform exhibited a notable switch in peptide binding specificity, with enhanced affinity for novel peptide sequences in either p53 or small nuclear ribonucleoprotein-U (snRNP-U) and substantially reduced affinity for the primary p53 binding site in the BOX-I domain. The consensus binding site for the RNA.MDM2 complex within p53 is SGXLLGESXF, which links the S9-S10 beta-sheets flanking the BOX-IV and BOX-V motifs in the core domain and which is a site of reversible conformational flexibility in p53. Mutation of conserved amino acids in the linker at Ser(261) and Leu(264), which bridges the S9-S10 beta-sheets, stimulated p53 activity from reporter templates and increased MDM2-dependent ubiquitination of p53. Furthermore, mutation of the conserved Phe(270) within the S10 beta-sheet resulted in a mutant p53, which binds more stably to RNA.MDM2 complexes in vitro and which is strikingly hyper-ubiquitinated in vivo. Introducing an Ala(19) mutation into the p53(F270A) protein abolished both RNA.MDM2 complex binding and hyper-ubiquitination in vivo, thus indicating that p53(F270A) protein hyper ubiquitination depends upon MDM2 binding to its primary site in the BOX-I domain. Together, these data identify a novel MDM2 binding interface within the S9-S10 beta-sheet region of p53 that plays a regulatory role in modulating the rate of MDM2-dependent ubiquitination of p53 in cells. PMID- 11925450 TI - Functional conservation of subfamilies of putative UDP-N acetylgalactosamine:polypeptide N-acetylgalactosaminyltransferases in Drosophila, Caenorhabditis elegans, and mammals. One subfamily composed of l(2)35Aa is essential in Drosophila. AB - The completed fruit fly genome was found to contain up to 15 putative UDP-N acetyl-alpha-d-galactosamine:polypeptide N-acetylgalactosaminyltransferase (GalNAc-transferase) genes. Phylogenetic analysis of the putative catalytic domains of the large GalNAc-transferase enzyme families of Drosophila melanogaster (13 available), Caenorhabditis elegans (9 genes), and mammals (12 genes) indicated that distinct subfamilies of orthologous genes are conserved in each species. In support of this hypothesis, we provide evidence that distinctive functional properties of Drosophila and human GalNAc-transferase isoforms were exhibited by evolutionarily conserved members of two subfamilies (dGalNAc-T1 (l(2)35Aa) and GalNAc-T11; dGalNAc-T2 (CG6394) and GalNAc-T7). dGalNAc-T1 and novel human GalNAc-T11 were shown to encode functional GalNAc-transferases with the same polypeptide acceptor substrate specificity, and dGalNAc-T2 was shown to encode a GalNAc-transferase with similar GalNAc glycopeptide substrate specificity as GalNAc-T7. Previous data suggested that the putative GalNAc transferase encoded by l(2)35Aa had a lethal phenotype (Flores, C., and Engels, W. (1999) Proc. Natl. Acad. Sci. U. S. A. 96, 2964-2969), and this was substantiated by sequencing of three lethal alleles l(2)35Aa(HG8), l(2)35Aa(SF12), and l(2)35Aa(SF32). The finding that subfamilies of GalNAc transferases with distinct catalytic functions are evolutionarily conserved stresses that GalNAc-transferase isoforms may serve unique biological functions rather than providing functional redundancy, and this is further supported by the lethal phenotype of l(2)35Aa. PMID- 11925451 TI - Is the conduction of the nerve impulse a chemical or a physical process? 1907. PMID- 11925452 TI - 2001 W.O. Atwater Memorial Lecture and the 2001 ASNS President's Lecture: Human nutrient requirements: the challenge of the post-genome era. PMID- 11925453 TI - Quercetin-3-glucoside is transported by the glucose carrier SGLT1 across the brush border membrane of rat small intestine. AB - In the present study we investigated a possible involvement of the intestinal sodium-dependent glucose transporter (SGLT)1 in the absorption of quercetin-3 glucoside (Q3G). Pieces of rat jejunum or proximal colon were mounted in Ussing type chambers and incubated under short-circuited conditions. Test flavonols were added to the mucosal or serosal bathing solution (initial concentration, 100 micromol/L) and disappearance from the donor compartment was monitored for 2 h. With jejunal tissue, only 13.6 +/- 3.5% of the initial dose of Q3G was found in the mucosal compartment 2 h after mucosal addition. Simultaneous addition of D glucose (10 mmol/L) significantly reduced the disappearance of Q3G (remaining concentration, 33.4 +/- 6.9%) as did a Na(+)-free buffer solution containing phloridzin (final mucosal concentration of Q3G, 54.2 +/- 7.7%). In these experiments, disappearance of Q3G was paralleled by the appearance of quercetin in the mucosal solutions. In contrast, D-fructose (10 mmol/L) did not influence the disappearance of Q3G (Na(+)-free conditions). With proximal colon, 78.2 +/- 11.5% of the initial concentration of Q3G was still present in the mucosal solution after 2 h. When added to the serosal side, the concentration of Q3G decreased only slightly (jejunum, 96.1 +/- 2.1%; proximal colon, 90.7 +/- 1.2%). The concentration of rutin did not change after mucosal or serosal addition. Neither transport of intact glycosides nor of free quercetin from the donor into the acceptor compartment was observed under our experimental conditions. Taken together, the results clearly indicate a role of SGLT1 in mucosal uptake of the Q3G. PMID- 11925454 TI - Acetyl-L-carnitine supplementation differently influences nutrient partitioning, serum leptin concentration and skeletal muscle mitochondrial respiration in young and old rats. AB - Variations in energy balance, body composition, and nutrient partitioning induced by acetyl-L-carnitine (ALCAR) supplementation were studied in young (2 mo) and old (24 mo) Wistar rats. Changes in skeletal muscle metabolism as well as in serum free triiodothyronine and leptin levels were also evaluated. Rats were administered 0 (control) or 15 g/L ALCAR in their drinking water for 1 mo. ALCAR treatment significantly decreased body lipid percentage in young rats and significantly increased body protein percentage in old rats. The percentage of metabolizable energy (ME) intake stored as lipid was lower in ALCAR-treated young rats, whereas the percentage of ME intake stored as protein was greater in ALCAR treated old rats compared with their age-matched controls. In addition, ALCAR supplementation significantly decreased serum leptin levels in old rats. Elevated skeletal muscle respiration was found in old rats treated with ALCAR, due to an increase in mitochondrial protein mass. In conclusion, ALCAR supplementation decreases efficiency of lipid deposition in young rats and increases efficiency of protein deposition in old rats. In addition, ALCAR supplementation partly reduces the leptin resistance that occurs in old rats, and improves ATP production in skeletal muscle mitochondria through an increase in mitochondrial protein content. PMID- 11925455 TI - Iron deficiency decreases mitochondrial aconitase abundance and citrate concentration without affecting tricarboxylic acid cycle capacity in rat liver. AB - Mitochondrial aconitase (m-acon) is the tricarboxylic acid (TCA) cycle enzyme that converts citrate to isocitrate. m-Acon mRNA is a potential target for regulation by iron regulatory proteins (IRPs), suggesting a link between dietary iron intake, m-acon synthesis, and energy metabolism. Our previous studies indicate that m-acon is one of a limited number of proteins that is down regulated in iron-deficient liver. Here we use isolated hepatocytes to study the relationships among decreased m-acon abundance, TCA cycle function and cellular citrate concentration in iron deficiency. Rats were fed an iron-deficient (ID) (2 mg Fe/kg diet) diet, or they were pair-fed (PF) or freely fed (C) a control diet (50 mg Fe/kg diet) for up to 21 d. Hepatocyte total IRP activity was greater by d 2 in the ID group than in the C and PF groups and by d 10, the difference was maximal. Liver IRP activity was inversely correlated with m-acon abundance (r = 0.93, P < 0.0001). However, the decrease in m-acon abundance did not affect the ability of hepatocytes to oxidize 2-[(14)C]pyruvate or 1-[(14)C]acetate, indicating that TCA cycle capacity was not affected. Interestingly, by d 21, total liver citrate concentration was 40% lower in ID than in PF rats, suggesting enhanced utilization of citrate. However, the decrease in citrate concentration was not reflected in a change in liver total lipid concentration. Taken together, our results indicate that the iron-dependent regulation of m-acon in liver does not alter TCA cycle capacity but suggest that IRP-mediated changes in m-acon expression may modulate citrate use in other aspects of intermediary or iron metabolism. PMID- 11925456 TI - Dietary nucleotide supplementation reduces thioacetamide-induced liver fibrosis in rats. AB - Dietary nucleotides reportedly promote functionality and repair in fibrotic liver. Liver fibrosis is characterized by an excessive accumulation of extracellular matrix components, which lead to the impairment of the hepatic function. The aim of this work was to evaluate the influence of dietary nucleotides on liver fibrosis induced by thioacetamide and to elucidate the mechanism by which nucleotides exert their protective effects. Rats consumed ad libitum 300 mg/L thioacetamide in drinking water and were pair-fed diets with (group TN) or without nucleotides (group TS) for 4 mo. Liver histology and extracellular matrix components, liver collagenase and prolyl 4-hydroxylase activities, and tissue inhibitor of metalloproteinases-1 were assessed. The degree of fibrosis was lower in group TN than in group TS. Group TN had lower hepatic concentration of hydroxyproline (P < 0.05), collagen type I (P = 0.12) and type III (P = 0.20), fibronectin (P = 0.05), laminin (P = 0.11) and desmin (P = 0.07), higher collagenolytic activity (P < 0.05), lower prolyl 4-hydroxylase activity (P < 0.05) and lower prolyl 4-hydroxylase (P = 0.10) and tissue inhibitor of metalloproteinase-1 (P = 0.06) expression than group TS. Moreover, expression of tissue inhibitor of the metalloproteinases-1 gene was lower in group TN than in group TS (P < 0.05). These data indicate that the reduction of liver fibrosis in nucleotide-supplemented rats may rely on the enhancement of collagenase activity and the reduction of collagen content and maturation. PMID- 11925457 TI - Phytoestrogens and mycoestrogens bind to the rat uterine estrogen receptor. AB - Consumption of phytoestrogens and mycoestrogens in food products or as dietary supplements is of interest because of both the potential beneficial and adverse effects of these compounds in estrogen-responsive target tissues. Although the hazards of exposure to potent estrogens such as diethylstilbestrol in developing male and female reproductive tracts are well characterized, less is known about the effects of weaker estrogens including phytoestrogens. With some exceptions, ligand binding to the estrogen receptor (ER) predicts uterotrophic activity. Using a well-established and rigorously validated ER-ligand binding assay, we assessed the relative binding affinity (RBA) for 46 chemicals from several chemical structure classes of potential phytoestrogens and mycoestrogens. Although none of the test compounds bound to ER with the affinity of the standard, 17beta-estradiol (E(2)), ER binding was found among all classes of chemical structures (flavones, isoflavones, flavanones, coumarins, chalcones and mycoestrogens). Estrogen receptor relative binding affinities were distributed across a wide range (from approximately 43 to 0.00008; E(2) = 100). These data can be utilized before animal testing to rank order estimates of the potential for in vivo estrogenic activity of a wide range of untested plant chemicals (as well as other chemicals) based on ER binding. PMID- 11925458 TI - Short-term zinc deficiency inhibits chondrocyte proliferation and induces cell apoptosis in the epiphyseal growth plate of young chickens. AB - The purpose of this study was to investigate the effect of zinc deficiency on chondrocyte proliferation, differentiation and apoptosis in the epiphyseal growth plate of juvenile chickens. Newly hatched broiler chickens were fed either a low zinc (10 mg/kg diet) or a zinc-adequate (68 mg/kg diet) soy protein-based purified diet. Cell proliferation in the growth plate was evaluated with bromodeoxyuridine (BrdU) labeling. Apoptosis was assessed using the terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling (TUNEL) method. Chondrocyte differentiation was evaluated with immunostaining of osteonectin as a marker of maturation. As early as d 3 of feeding, zinc deficiency significantly inhibited chondrocyte proliferation, promoted cell differentiation and induced cell apoptosis in the growth plate. These effects were manifested primarily in areas remote from the blood supply. Immunostaining for local growth factors such as insulin-like growth factor-1 (IGF-1), parathyroid hormone-related protein (PTHrP) and fibroblast growth factor-2 (FGF-2) did not reveal any differences between growth plates of zinc-deficient and zinc-adequate chickens after 3 d of feeding. By d 7, severe growth plate lesions characterized by reduced cellularity and abnormally shaped cells were formed in areas remote from blood vessels. Immunoreactive IGF-1, PTHrP and FGF-2 were all greatly reduced in the lesion. However, the growth rate and food intake of zinc-deficient chickens were not different from those of the controls during the 7-d experiment. Therefore, a direct effect of zinc deficiency on proliferation, differentiation, and apoptosis of growth plate chondrocytes was indicated. PMID- 11925459 TI - Selenite and selenomethionine promote HL-60 cell cycle progression. AB - The essential role of selenium (Se) in nutrition is well established. The elucidation of the mechanisms by which selenium regulates the cell cycle can lead to a better understanding of the nature of selenium's essentiality and its role in disease prevention. In this study, the effects of selenium deficiency or adequacy (0.25 micromol/L selenite or selenomethionine) on HL-60 cell cycle progression were examined in serum-free media. Selenium was critical for promotion of HL-60 cell growth. Cell-cycle analysis revealed that selenium deficiency caused a decrease in G1 phase cells that corresponded to an increase in G2 and sub-G1 phase cells. Gene array analysis suggested that c-Myc, cyclin C, proliferating cell nuclear antigen, cyclin-dependent kinase (cdk)1, cdk2, cdk4, cyclin B and cyclin D2 mRNA levels were lower in selenium-deficient cells than in the cells supplemented with 0.25 micromol/L selenomethionine. The decrease in the c-Myc mRNA level in selenium-deficient cells was confirmed by reverse transcription-polymerase chain reaction analysis. Furthermore, the phosphorylation state of total cellular protein was higher (57%) in selenium supplemented cells than in selenium-deficient cells. Collectively, these results suggest a novel role for selenium at 0.25 micromol/L in up-regulation of the expression of numerous cell cycle-related genes and total cellular phosphorylated proteins in HL-60 cells in serum-free culture media. This leads to the promotion of cell cycle progression, particularly G2/M transition and/or the reduction of apoptosis, primarily in G1 cells. These observations may have additional implications for understanding the nature of selenium's essentiality. PMID- 11925460 TI - IEC-6 cells are an appropriate model of intestinal iron absorption in rats. AB - Regulation of iron absorption, which is the primary mechanism for maintaining body iron stores, occurs primarily in the proximal small intestine. Recent identification of proteins that are involved in iron absorption such as the uptake transporter-divalent metal transporter (DMT1), the basolateral transporter, IREG1, and the ferroxidase-hephaestin provide new opportunities to study this process. We evaluated the rat intestinal cell line, IEC-6, as a model of intestinal iron transport. This involved measuring the expression of DMT1 and IREG1 by Western blot analysis and confocal microscopy, and hephaestin by protein dependent copper oxidase activity. DMT1 and IREG1 were expressed in IEC-6 cells. The uptake of 1 micromol/L ferrous iron [Fe(II)]:ascorbate and its efflux also was associated with the expression of DMT1 under different levels of iron loading. The expression of DMT1 changed inversely with iron levels as did the uptake of Fe(II). However, with different levels of cellular iron, IREG1 expression remained constant, as did the release of iron from the cells, suggesting that they could be related. Ceruloplasmin and apotransferrin did not enhance the rate or extent of iron release. Copper oxidase activity, considered to indicate hephaestin activity, was detected only intracellularly. Confocal microscopy showed DMT1 and IREG1 on the cell membrane of IEC-6 cells at 4 degrees C but at intracellular locations at 37 degrees C, indicating that these proteins can function at the cell membrane and intracellularly. In terms of iron absorption, IEC-6 cells have a villous enterocyte phenotype and are regulated by iron stores as occurs in vivo; therefore, they represent an appropriate cell model with which to study this process. PMID- 11925461 TI - Differential regulation of porcine hepatic IGF-I mRNA expression and plasma IGF-I concentration by a low lysine diet. AB - The influence of dietary lysine on hepatic insulin-like growth factor-I (IGF-I) gene expression and plasma IGF-I level was investigated. Two male 6-wk-old pigs from each of six litters were used. Each littermate was assigned to one of two diets, control or low lysine (LL), that were isoenergetic and similar in protein content and provided 14.3 MJ digestible energy/kg for both diets, 185 g protein/kg for the control diet and 180 g protein/kg for the LL diet. The control diet contained all essential amino acids in the recommended amounts, including 11.5 g lysine/kg. The LL diet was similar but contained only 7 g lysine/kg. Pigs were pair-fed these diets for 3 wk. Growth rates and feed efficiencies of pigs fed the LL diet were significantly lower than those of pigs fed the control diet (P < 0.01). Plasma IGF-I levels in pigs fed the LL diet were 52% lower than in those fed the control diet (P < 0.01), and the LL group also had lower plasma IGF binding protein-3 (IGFBP3) levels (P < 0.05). Despite the strikingly lower plasma IGF-I in pigs fed the LL diet, hepatic IGF-I mRNA abundance did not differ between the two treatment groups. We conclude that the reduction in plasma IGF-I caused by reduced dietary lysine may have been due in part to suppression of post transcriptional events in IGF-I expression. The lower plasma IGFBP3 in pigs fed the LL diet suggests that increased clearance rates of circulating IGF-I may have been involved in this response. PMID- 11925462 TI - Iron treatment downregulates DMT1 and IREG1 mRNA expression in Caco-2 cells. AB - Iron deficiency is the most common nutritional disorder worldwide, whereas pathologic elevations of body iron stores can occur under certain circumstances due to genetic abnormalities or in association with other diseases. The intestine is the exclusive locus of homeostatic regulation of body iron stores, which is accomplished by changes in iron absorption efficiency by largely unknown molecular mechanisms in response to alterations in body iron stores. Recently, a number of novel genes involved in iron metabolism, such as the iron uptake transporter DMT1/DCT1/Nramp2 and the iron export transporter IREG1/ferroportin1/MTP1, have been identified, providing important insights about molecular aspects of intestinal iron absorption and its regulation. The aim of this study was to investigate the effects of iron treatment on DMT1 and IREG1 mRNA expression in Caco-2 cells, a human intestinal cell line. Exposure of the cells to iron (200 micromol/L ferric nitrilotriacetic acid for 72 h) significantly decreased transferrin receptor mRNA (80%), DMT1 mRNA (57%) and IREG1 mRNA (52%). These observations are consistent with the notion of parallel regulation of these iron-responsive genes in vivo to protect the enterocyte from iron toxicity and mediate a decreased efficiency of intestinal iron absorption to prevent iron overload. PMID- 11925463 TI - The effect of five-year zinc supplementation on serum zinc, serum cholesterol and hematocrit in persons randomly assigned to treatment group in the age-related eye disease study: AREDS Report No. 7. AB - The effects of long-term supplementation with pharmacologic doses of zinc oxide on serum levels of zinc, lipids and hematocrit have not been studied systematically to date. Eleven Clinical Centers enrolled 4757 participants from 1992 to 1998 as part of the Age-Related Eye Disease Study (AREDS). Of these, 3640 participants, aged 55-80 y, who had early-to-late age-related macular degeneration (AMD) were randomly assigned to daily supplementation with or without 80 mg of zinc as zinc oxide plus 2 mg of copper as cupric oxide to study the effects of zinc supplementation on the progression to late AMD. This paper reports on the effect of a 5-y supplementation with zinc oxide and cupric oxide on serum zinc, copper, lipids, and hematocrit for 717 participants from three clinical centers. At the 5-y exam, the median increase in serum zinc levels for participants assigned to zinc formulations was 17% compared with a 2% increase for participants not assigned to zinc (P < 0.001). The differential effect on serum zinc was observed at 1 y and remained fairly constant over the 5-y period. After 5 y, no significant differences in changes in serum hematocrit, copper or lipids were found between participants assigned to formulations containing zinc and copper, and those assigned to formulations without zinc and copper. Estimates from a modified Block Food-Frequency Questionnaire suggest the AREDS population at baseline had a zinc intake from diet similar to that of the general population. PMID- 11925464 TI - Almonds and almond oil have similar effects on plasma lipids and LDL oxidation in healthy men and women. AB - Epidemiologic and clinical studies have shown that nut consumption is associated with favorable plasma lipid profiles and reduced cardiovascular risk. These effects may result from their high monounsaturated fat (MUFA) content but nuts contain constituents other than fatty acids that might be cardioprotective. We conducted a study to compare the effects of whole-almond vs. almond oil consumption on plasma lipids and LDL oxidation in healthy men and women. Using a randomized crossover trial design, 22 normolipemic men and women replaced half of their habitual fat (approximately 14% of approximately 29% energy) with either whole almonds (WA) or almond oil (AO) for 6-wk periods. Compliance was ascertained by monitoring dietary intake via biweekly 5-d food records, return of empty almond product packages and weekly meetings with a registered dietitian. Fat replacement with either WA and AO resulted in a 54% increase in percentage of energy as MUFA with declines in both saturated fat and cholesterol intake and no significant changes in total energy, total or polyunsaturated fat intake. The effects of WA and AO on plasma lipids did not differ compared with baseline; plasma triglyceride, total and LDL cholesterol significantly decreased, 14, 4 and 6% respectively, whereas HDL cholesterol increased 6%. Neither treatment affected in vitro LDL oxidizability. We conclude that WA and AO do not differ in their beneficial effects on the plasma lipid variables measured and that this suggests that the favorable effect of almonds is mediated by components in the oil fraction of these nuts. PMID- 11925465 TI - Effects of soy consumption on gonadotropin secretion and acute pituitary responses to gonadotropin-releasing hormone in women. AB - Soy contains the isoflavone phytoestrogens, genistein and daidzein. These isoflavones are partial estrogen agonists in cell and animal models, but effects from dietary soy in humans are unclear. Experiments were conducted in pre- and postmenopausal women to examine whether dietary isoflavones from soy behave as estrogen agonists, antagonists or have no effect on the estrogen-sensitive pituitary. Pituitary sensitivity to gonadotropin-releasing hormone (GnRH), an estrogen-sensitive endpoint, was measured during GnRH challenge tests administered before, during and after dietary soy consumption. The response to an isoflavone-rich soy food diet was examined in five premenopausal and seven postmenopausal women using transdermal estrogen replacement therapy. Estrogen agonists suppress gonadotropin concentrations and enhance GnRH priming (enhanced gonadotropin secretion in response to repeated doses of GnRH), whereas antagonists elevate gonadotropin concentrations and have no effect on GnRH priming. Each subject consumed 50 g textured soy protein containing 60 mg total isoflavones daily for 10-14 d. Baseline estradiol concentrations were consistent among study periods. In both pre- and postmenopausal women, soy consumption did not affect mean baseline or peak luteinizing hormone (LH) concentrations, indicating a lack of estrogen-like effect at the level of the pituitary. However, in postmenopausal subjects, mean LH secretion decreased after discontinuing soy, suggesting a residual estrogenic effect. In one premenopausal woman, enhanced LH secretion was observed after soy treatment, suggesting there may be subpopulations of women who are highly sensitive to isoflavones. PMID- 11925466 TI - Dietary mono- and polyunsaturated fatty acids similarly affect LDL size in healthy men and women. AB - The goal of this study was to investigate the effect of the dietary fat composition on LDL peak particle diameter. Therefore, we measured LDL size by gradient gel electrophoresis in 56 (30 men, 26 women) healthy participants in a controlled dietary study. First, all participants received a baseline diet rich in saturated fat for 2 wk; they were then randomly assigned to one of three dietary treatments, which contained refined olive oil [rich in monounsaturated fatty acids (MUFA), n = 18], rapeseed oil [rich in MUFA and (n-3)-polyunsaturated fatty acids (PUFA), n = 18], or sunflower oil [rich in (n-6)-PUFA, n = 20] as the principal source of fat for 4 wk. Repeated-measures ANOVA revealed a small, but significant reduction in LDL size during the oil diet phase (-0.36 nm, P = 0.012), which did not differ significantly among the three groups (P = 0.384). Furthermore, affiliation with one of the three diet groups did not contribute significantly to the observed variation in LDL size (P = 0.690). In conclusion, our data indicate that dietary unsaturated fat similarly reduces LDL size relative to saturated fat. However, the small magnitude of this reduction also suggests that the composition of dietary fat is not a major factor affecting LDL size. PMID- 11925468 TI - The composition of the major molecular species of adipose tissue triacylglycerols of rats reflects those of dietary rapeseed, olive and sunflower oils. AB - We report the composition of constituent fatty acids and molecular species of adipose tissue triacylglycerols of male weaned Wistar rats fed diets containing, in addition to 20 g corn oil/kg feed, 120 g per kg feed canola-type rapeseed oil, olive oil or conventional sunflower oil for 10 wk. The composition of fatty acids and molecular species of the triacylglycerols of subcutaneous, epididymal and perirenal adipose tissues did not differ among groups (P > 0.01), broadly reflecting the corresponding compositions of the dietary oils. The major molecular species of dietary triacylglycerols, especially trioleoylglycerol (OOO) and linoleoyl-dioleoylglycerols (LOO) in the rapeseed oil and olive oil diets, dioleoyl-palmitoylglycerols (OOP) in the olive oil diet, dilinoleoyl oleoylglycerols (LLO) in the rapeseed oil and sunflower oil diets, and dilinoleoyl-palmitoylglycerols (LLP), linoleoyl-oleoyl-palmitoylglycerols (LOP) as well as trilinoleoylglycerol (LLL) in the sunflower oil diet were also prominent constituents of the corresponding adipose tissue triacylglycerols. On the other hand, predominant molecular species containing alpha-linolenoyl (Ln) moieties, e.g., alpha-linolenoyl-linoleoyl-oleoylglycerols (LnLO) and alpha linolenoyl-dioleoylglycerols (LnOO) from the rapeseed oil diet were not prominent constituents of rat adipose tissue triacylglycerols, whereas LOP from rapeseed oil and olive oil diets and OOP from rapeseed oil and sunflower oil diets were distinctly enriched in the corresponding adipose tissues. Most of the minor molecular species of the dietary triacylglycerols from all the three diets were distinctly present in the corresponding adipose tissues. Thus, despite numerous biochemical processes involved in the metabolism of dietary triacylglycerols, a substantial proportion of the molecular species of adipose tissue triacylglycerols containing linoleoyl (L), oleoyl (O) and palmitoyl (P) moieties resemble those of dietary triacylglycerols. PMID- 11925467 TI - Family food insufficiency, but not low family income, is positively associated with dysthymia and suicide symptoms in adolescents. AB - Food insufficiency has been shown to be associated with poor health, academic and psychosocial outcomes in American children, but the relationship between food insufficiency and depressive disorders in U.S. adolescents has not been studied. Further, there are no national estimates of the prevalence of depressive disorders for U.S. adolescents, nor investigation of associations with sociodemographic characteristics using national data. Therefore, we analyzed data for 15- and 16-y-old adolescents from the Third National Health and Nutrition Examination Survey (NHANES III). Depressive disorders and suicidal symptoms were assessed using the Diagnostic Interview Schedule. Adolescents were classified as "food insufficient" if a family respondent reported that the family sometimes or often did not have enough to eat. The prevalence of depression outcomes is reported by sociodemographic characteristics. Odds ratios for associations with food insufficiency are reported, adjusted for sociodemographic factors. Overall, lifetime prevalence of major depressive disorder was 6.3% and of dysthymia, 5.4%. Almost 5% of 15- to 16-y-old adolescents reported that they had ever attempted suicide and 38.8% reported at least one suicidal symptom. Female adolescents were significantly more likely than males to have had dysthymia, any depressive disorder and all symptoms of suicide. Low income adolescents were less likely to report suicide ideation than high income adolescents, but there were no other differences by family income. Food-insufficient adolescents were significantly more likely to have had dysthymia, thoughts of death, a desire to die and have attempted suicide. There is a strong association between food insufficiency and depressive disorder and suicidal symptoms in U.S. adolescents. PMID- 11925469 TI - L-type amino acid transporters in two intestinal epithelial cell lines function as exchangers with neutral amino acids. AB - The present study examined the functional characteristics of the inward [(14)C]-L leucine transporter in two intestinal epithelial cell lines (human Caco-2 and rat IEC-6). The uptake of [(14)C]-L-leucine was largely promoted through an energy dependent and sodium-insensitive transporter, although a minor component of [(14)C]-L-leucine uptake ( approximately 15%) required extracellular sodium. [(14)C] -L-leucine uptake was insensitive to N-(methylamino)-isobutyric acid, but competitively inhibited by 2-aminobicyclo(2,2,1)-heptane-2-carboxylic acid (BCH). Both L- and D-neutral amino acids, but not acidic and basic amino acids, markedly inhibited [(14)C]-L-leucine accumulation. The efflux of [(14)C]-L-leucine was markedly increased (P < 0.05) by L-leucine and BCH, but not by L-arginine. In IEC 6 cells, but not in Caco-2 cells, the uptake of [(14)C]-L-leucine at acidic pH (5.0 and 5.4) was greater (P < 0.05) than at pH 7.4. In conclusion, it is likely that system B(0) might be responsible for the sodium-dependent uptake of L leucine in Caco-2 and IEC-6 cells, whereas sodium-independent uptake of L-leucine may include system LAT1, whose activation results in transstimulation of L leucine outward transfer. PMID- 11925470 TI - Energy metabolism and turnover are increased in mice lacking the cholecystokinin B receptor. AB - Cholecystokinin (CCK) is an important gastrointestinal hormone as well as a neurotransmitter. Two types of CCK receptors, types A and B, have been identified. The CCK-A receptor is involved in satiety, food intake and behavior, whereas the B receptor is involved in anxiety. We recently produced CCK-A, -B and AB receptor knockout mice to study the role of these receptors in energy metabolism. Daily energy intake and expenditure were significantly greater in CCK BR(-/-) and CCK-AR(-/-)BR(-/-) mice than CCK-AR(-/-) and wild-type [CCK AR(+/+)BR(+/+)] mice. Relative liver and kidney weights (g/kg body) were significantly greater in CCK-AR(-/-)BR(-/-) mice than in wild-type mice. Energy metabolism and energy turnover were increased in mice with a disruption of the CCK-BR gene, although the underlying mechanism is unknown. PMID- 11925471 TI - Dietary fish oil and Undaria pinnatifida (wakame) synergistically decrease rat serum and liver triacylglycerol. AB - Japanese eating habits are characterized by the consumption of various food materials such as cereals, vegetables, fish, shellfish, marine algae and meat. Therefore, properties of functional substances in food materials may be enhanced or lessened by the combination of various food materials. In the present study, we examined how the combination of wakame and fish containing polyunsaturated fatty acids, which are typical Japanese food materials, affected rat lipid metabolism. Rats were fed one of four diets [control diet (C), AIN-76 diet with 5 g/100 g rapeseed oil; wakame diet (W) containing 19.1 g/100 g Undaria pinnatifida (wakame) dried powder in the C diet; fish oil diet (FO), AIN-76 diet with 4.1 g/100 g fish oil; wakame-fish oil diet (W + FO), the FO diet containing 19.1 g/100 g dried wakame powder] for 4 wk. We measured the concentration of lipids in serum and liver and hepatic activities of enzymes involved in fatty acid metabolism. The W diet, FO diet and W + FO diet significantly reduced the concentration of triacylglycerols in the serum and liver compared with the C diet. This decrease in the concentration of hepatic triacylglycerol was greatest in rats fed the W + FO diet. The activity of glucose-6-phosphate dehydrogenase, which is involved in fatty acid synthesis in the liver, of rats fed the W, FO and W + FO diets was lower than that in rats fed the C diet. However, the activities of malic enzyme and fatty acid synthetase did not differ among the four groups. In contrast, the W diet and W + FO diet increased the serum concentration of beta hydroxybutyrate. Further, the activity of 3-hydroxyacyl-CoA dehydrogenase, which is involved in fatty acid beta-oxidation in the liver, was greater in rats fed the W diet (42%), the FO diet (154%) and the W + FO diet (381%) than in those fed the C diet. Because the decrease in the concentration of triacylglycerol in the liver was greatest when rats were fed wakame and fish oil at the same time (W + FO diet), we conclude that there was a synergistic process affecting fatty acid beta-oxidation in the liver. These results suggest that the simultaneous consumption of fish (fish oil) and wakame decreases the concentration of triacylglycerol in the serum and liver. PMID- 11925472 TI - Dietary vitamin C and vitamin E interact to influence growth and tissue composition of juvenile hybrid striped bass (Morone chrysops (female) x M. saxatilis (male)) but have limited effects on immune responses. AB - Juvenile hybrid striped bass (initially 12.0 g) were fed diets containing deficient, adequate or excessive amounts of vitamin C and/or vitamin E in a factorial arrangement to investigate potential nutritional interaction and effects on immune responses. Nine semipurified diets were supplemented with 0, 25 or 2500 mg vitamin C/kg and 0, 30 or 300 mg vitamin E/kg and fed to fish in triplicate aquaria for 10 wk. Weight gain, feed efficiency, mortality and tissue vitamin levels were significantly (P < or = 0.05) affected by dietary vitamin levels. In addition, a significant interaction between vitamin C and vitamin E was observed. At inclusion levels of 25 and 2500 mg/kg, dietary vitamin C improved feed efficiency and protected fish fed vitamin E-deficient diets from growth depression and mortality. At inclusion levels of 30 and 300 mg/kg, vitamin E prevented mortality in fish fed vitamin C-deficient diets; however, 300 mg vitamin E/kg was necessary to prevent growth depression in vitamin C-deficient fish but was unable to improve feed efficiency. Lysozyme, bacterial killing ability, as well as plasma protein and total immunoglobulin levels of fish were not affected by dietary vitamin levels, whereas respiratory burst activity increased with vitamin E supplementation. Thus, interactions between vitamin C and vitamin E were observed in hybrid striped bass. These interactions may be due to the ability of vitamin C to regenerate vitamin E to its functional form but also suggest an ability of vitamin E to spare vitamin C. PMID- 11925473 TI - A dietary oxidative balance score of vitamin C, beta-carotene and iron intakes and mortality risk in male smoking Belgians. AB - The purpose of this study was to investigate, in smokers, whether the oxidative balance of their dietary pattern affected mortality risk. To evaluate the oxidative balance of the dietary pattern, an oxidative balance score was constructed that summarized the combined intake of dietary antioxidants (vitamin C and beta-carotene) and a prooxidant (iron). The low oxidative balance score group included smokers with a diet high in vitamin C and beta-carotene and/or low in iron and the high oxidative balance score group included those with a diet low in vitamin C and beta-carotene and/or high in iron. Using the 10-y follow-up mortality data from the Belgian Interuniversity Research on Nutrition and Health (BIRNH) study, the association of this oxidative balance score with all-cause, cardiovascular disease (CVD) and total cancer mortality was investigated in 2814 male smokers. In multivariate-adjusted Cox models, men in the highest oxidative balance score group had a higher relative risk (RR) of all-cause [RR = 1.44, 95% confidence interval (CI): 1.13, 1.82] and of total cancer mortality (RR = 1.62, 95% CI: 1.07, 2.45) compared with men in the lowest score group. This association was less pronounced for CVD mortality risk and was not significant (RR = 1.31, 95% CI: 0.86, 2.00). The risk of all-cause and total cancer mortality was driven principally by the high score group, which suggested a threshold effect for risk rather than a linear trend. The findings are consistent with the hypothesis that the oxidative balance of the diet is associated with subsequent mortality. Smokers whose diet is unbalanced in terms of anti- and prooxidants may therefore benefit from a recommendation to consume more servings of fresh fruits and vegetables and less meat. PMID- 11925474 TI - Folate intake, serum homocysteine and methylenetetrahydrofolate reductase (MTHFR) C677T genotype are not associated with oral cancer risk in Puerto Rico. AB - We examined the relationships between folate and methionine intake, serum homocysteine levels (as a biomarker for folate metabolism), and methylenetetrahydrofolate reductase (MTHFR) C677T polymorphism genotype and risk of oral cancer in a population-based, case-control study in Puerto Rico. Structured questionnaires were used to collect information on demographic factors, usual adult diet, and tobacco and alcohol use. Oral epithelial cells and blood samples were collected from a subset of subjects. Analyses were conducted by logistic regression, adjusting for age, sex, lifetime smoking and lifetime alcohol intake, with the following numbers of cases/controls, respectively: dietary data (341/521); MTHFR genotype (148/149); and homocysteine (60/90). Although increased folate intake was associated with decreased oral cancer risk [adjusted odds ratio (OR) in highest vs. lowest quartile = 0.6, 95% confidence interval (CI): 0.4, 1.0, P(trend) = 0.05)], this finding was due almost entirely to folate intake from fruit (adjusted OR = 0.4, 95% CI: 0.2, 0.6; P(trend) = 0.0001), whereas other dietary folate sources showed no clear association. Methionine intake and serum homocysteine levels were not associated with oral cancer risk. Subjects with the MTHFR C677T homozygous variant (TT) genotype had a nonsignificantly lower risk, and risk patterns tended to differ by level of folate, methionine, alcohol intake and smoking, although the power to detect significant associations in subgroups of these variables was low. Risks for oral cancer are not folate specific; preventive recommendations for this disease should emphasize the importance of a healthy diet, including substantial intake of fruits. PMID- 11925475 TI - Food label use modifies association of income with dietary quality. AB - We analyzed the 1994-1996 Continuing Survey of Food Intake by Individuals and the Diet and Health Knowledge Survey (DHKS) to examine whether the relationship between income and dietary quality is modified by food label (FL) use among 20- to 60-y old DHKS respondents who were either household meal preparers, meal planners or food shoppers (n = 2952). Multivariate logistic regression results indicated that the influence of income on dietary quality is mediated by FL use. Those who were wealthier and used FL were significantly less likely to have a lower Healthy Eating Index (HEI) compared with the reference group formed by those in the lower income category who did not use FL [OR = 0.42; 95% confidence interval (CI): 0.31, 0.56]. By contrast, those who were wealthier but did not use FL were as likely as the reference group to have a low HEI (OR = 1.08; 95% CI: 0.74, 1.54). Those who were poorer but used FL were significantly less likely to have a low HEI compared with the reference group (OR = 0.62; 95% CI: 0.48, 0.80). Thus, FL use is associated with improved dietary quality among all income groups with a greater benefit of use among higher income individuals. Income is not associated with improved dietary quality in the absence of FL use. PMID- 11925476 TI - Diallyl disulfide (DADS) induces the antitumorigenic NSAID-activated gene (NAG-1) by a p53-dependent mechanism in human colorectal HCT 116 cells. AB - Garlic is appealing as an anti-carcinogenic agent due to its ability to induce apoptosis in vitro and inhibit the formation and growth of tumors in animals in vivo. Diallyl disulfide (DADS) is a constituent of garlic that suppresses neoplastic cell growth and induces apoptosis. We examined the effects of DADS on various cancer cell lines to better understand its effect on apoptosis and apoptosis-related genes. The nonsteroidal anti-inflammatory drug (NSAID) activated gene (NAG-1) has proapoptotic and antitumorigenic activities and is upregulated by anticancer agents such as NSAIDs. In this study, human colorectal HCT-116 (wild-type p53), HCT-15 (p53 mutant) and human prostate PC-3 (p53 mutant) cells were exposed to DADS. DADS inhibited cell proliferation in all cell lines albeit to a lesser extent in HCT-15 and PC-3 cells at 11.5 and 23 micromol/L. In HCT-116 cells, DADS induced p53 and NAG-1 in a dose-dependent manner and the induction of p53 preceded that of NAG-1. In HCT-116 cells, NAG-1 protein expression was increased 2.4-fold +/- 0.6 at 4.6 micromol/L and 6.1-fold +/- 1.7 at 23 micromol/L DADS, whereas p53 was induced 1.5-fold +/- 0.1 and 2.3-fold +/- 0.4. DADS did not induce NAG-1 or p53 in p53 mutant cell lines; however, NAG-1 expression was induced by sulindac sulfide. HCT-116 cells treated with 4.6 and 23 micromol/L DADS resulted in a 1.9- and 2.9-fold increase in apoptosis, respectively. In contrast, 23 micromol/L DADS induced apoptosis only 1.8-fold in HCT-15 cells and not at all in PC-3 cells. Thus, DADS-induced apoptosis and NAG-1 protein expression appear to occur via p53. PMID- 11925477 TI - Securing the future of nutritional sciences through integrative graduate education. AB - The new millennium has ushered in unprecedented opportunities for research and discovery in the nutritional sciences. These have arisen from major advancements in the social and biological sciences, notably genetics, genomic technologies, computational statistics, and behavioral sciences. The most exciting academic challenges and employment opportunities for nutritional sciences graduates interface with other disciplines, and doctoral training programs in the nutritional sciences must respond to the wealth of these emerging opportunities. The American Society for Nutritional Sciences (ASNS) Graduate Nutrition Education Committee suggests that three major challenges face the nutritional sciences: (a) to train doctoral students to be full and active participants in interdisciplinary research and discovery, and (b) to achieve this goal while maintaining nutrition's unique academic identity and (c) fostering a cohesive academic community. The committee proposes that these objectives are best engaged at the level of graduate education and that the future of ASNS will be secured by strengthening graduate nutrition programs. This manuscript reviews educational strategies that address these challenges and advocates that ASNS actively engage graduate doctoral programs in nutritional sciences. PMID- 11925478 TI - Antioxidant activity of black tea vs. green tea. PMID- 11925482 TI - Forging effective strategies to combat iron deficiency. Proceedings of the 6th Biennial International Conference on Health Promotion. May 7-9, 2001, Atlanta, Georgia, USA. PMID- 11925483 TI - Keynote address: issues in overcoming iron deficiency. PMID- 11925484 TI - Reversing productivity losses from iron deficiency: the economic case. AB - Iron deficiency anemia (IDA) depresses human productivity, but policymakers do not generally view this effect as an impediment to sustained economic growth. Economic logic should be enfolded in public advocacy for increased investment in the prevention and control of IDA. This paper argues that integrated strategies are required, with each intervention clearly related to particular groups at risk, and benefits carefully calibrated with costs. Protecting women's lives through supplementation has the first call on public resources, but the most productive investments reduce population prevalence at least cost, and these lie with food-based approaches within the competence of the processed food and seed industries. The public and private sectors must embark on modernization of the food industry in developing countries and reorientation of the international agricultural research complex so that iron-enriched essential foods will be affordable and accessible to the poor, especially children. The costs of IDA, the availability of cost-effective strategies and the benefits of sustained iron nutrition improvement to individuals, families and nations are reviewed. The roles of iron supplementation, food fortification, plant breeding and biotechnology, both actual and imminent, are described. The paper concludes that a recast Green Revolution directed toward dietary quality may be the key to enhancing the learning and earning capacity of young people in the developing world. PMID- 11925485 TI - Prevention and control of iron deficiency: policy and strategy issues. AB - Substantial efforts have been made in the past several decades to implement programs to reduce iron deficiency. Yet, compared with other micronutrients such as vitamin A and iodine, overall progress in reducing iron deficiency has been limited. Such limited progress is not attributed to a lack of scientific knowledge about the prevalence, causes or consequences of iron deficiency, but to limited implementation of effective interventions and ineffective communication tools. The challenge is to coordinate and balance research efforts more constructively with the implementation of practical and effective intervention programs. More attention must be paid to evaluating the operational feasibility of various intervention strategies to demonstrate their effectiveness under normal field conditions. Moreover, intervention efforts must be supported by substantially increased attention to communications to achieve effective advocacy for policy support and resource mobilization, foster partnerships and alliances, clarify priority target groups, including infants and young children, and support behavioral change. Through collaboration, researchers, program implementers and communicators can achieve substantial progress in reducing iron deficiency. PMID- 11925486 TI - Fortification: overcoming technical and practical barriers. AB - The main barriers to successful iron fortification are the following: 1) finding an iron compound that is adequately absorbed but causes no sensory changes to the food vehicle; and 2) overcoming the inhibitory effect on iron absorption of dietary components such as phytic acid, phenolic compounds and calcium. These barriers have been successfully overcome with some food vehicles but not with others. Iron-fortified fish sauce, soy sauce, curry powder, sugar, dried milk, infant formula and cereal based complementary foods have been demonstrated to improve iron status in targeted populations. The reasons for this success include the use of soluble iron such as ferrous sulfate, the addition of ascorbic acid as an absorption enhancer or the use of NaFeEDTA to overcome the negative effect of phytic acid. In contrast, at the present time, it is not possible to guarantee a similar successful fortification of cereal flours or salt. There is considerable doubt that the elemental iron powders currently used to fortify cereal flours are adequately absorbed, and there is an urgent need to investigate their potential for improving iron status. Better absorbed alternative compounds for cereal fortification include encapsulated ferrous sulfate and NaFeEDTA, which, unlike ferrous sulfate, do not provoke fat oxidation of cereals during storage. Encapsulated compounds also offer a possibility to fortify low grade salt without causing off-colors or iodine loss. Finally, a new and useful additional approach to ensuring adequate iron absorption from cereal based complementary foods is the complete degradation of phytic acid with added phytases or by activating native cereal phytases. PMID- 11925487 TI - Iron supplements: scientific issues concerning efficacy and implications for research and programs. AB - Iron supplementation remains an important strategy for the prevention and treatment of iron deficiency anemia and can produce substantial improvements in the functional performance of iron deficient individuals and populations. Many potential benefits of iron supplementation require further exploration, including its effect on vitamin A and iodine metabolism. There is strong evidence that vitamin A and riboflavin deficiencies affect iron utilization from supplements and are important on a global scale, but little evidence that folate and vitamin B-12 deficiencies play a major causal role in the global burden of anemia. The efficacy of multiple micronutrient supplements for the prevention and treatment of anemia must be further evaluated. Because weekly supplementation with iron is effective at improving iron status, this option should be thoroughly explored and evaluated in the context of programs for the prevention and the treatment of iron deficiency and anemia. More conformation is warranted concerning the number of tablets that must be consumed in different situations, and the efficacy of supplying other micronutrients weekly with iron. Weekly supplementation programs may improve the logistical and economic constraints that currently limit the provision of supplements to the many target population groups for whom they are recommended, but usually fail to reach. Further work is required to clarify the purpose, delivery and outcomes of iron supplementation programs. PMID- 11925488 TI - Experiences and challenges in industrialized countries: control of iron deficiency in industrialized countries. AB - This paper provides a synopsis of the experience in combating iron deficiency in industrialized countries and identifies the reasons for the considerable success and future challenges. Significant progress has been made over the last century in reducing and even eliminating iron deficiency in many industrialized countries. Current estimates are that the prevalence of iron deficiency has declined to <20% in many of these countries, even among women and young children, compared with 30 to 70% in many developing countries. The reasons for this success cannot be attributed solely to a single approach but rather to a range of factors that have occurred over time as a result of both economic development and the implementation of specific policies. Several factors have contributed to improving both iron intakes and reducing iron losses; these include fortification, supplementation, dietary diversification and public health measures. For example, the decline in anemia in infants can be attributed to the introduction of iron-fortified formula and complementary foods in the 1960s to 1970s. Similarly, the enrichment and fortification of cereals with iron that began during World War II in North America and Europe is a result of effective public-private partnerships. Despite these successes, iron deficiency remains a public health concern in industrialized countries for selected subgroups such as women of reproductive age with excess menstrual losses and pregnant women who cannot meet increased requirements from the diet alone. Constant vigilance and innovative approaches for screening and combating this problem are thus still required even in developed countries. PMID- 11925489 TI - Commentary: experiences and challenges in industrialized countries. AB - Industrialized nations have been successful in reducing incidences of iron deficiency through utilization of multifaceted strategies that build on the capabilities of successful alliances between the public and private sectors. In comparison to developing nations, which have an estimated prevalence of iron deficiency of approximately 40%, the prevalence in industrialized countries is approximately 5%. Such low rates were achieved by various initiatives including widespread availability of affordable iron-fortified foods and implementation of national, science-based programs. Despite these successes, however, iron deficiency remains a critical problem among certain subpopulations (e.g., people of low socioeconomic status or recent immigrants who do not consume iron fortified foods). Strengthening alliances among the private and public sectors and academia in communications efforts about iron deficiency and promoting iron rich foods as well as iron-fortified foods is necessary to reduce iron deficiency in these populations. PMID- 11925490 TI - Experiences and challenges in developing countries. AB - Compared with the industrialized nations, the challenges of combating iron (Fe) deficiency in developing countries include the far greater magnitude of the problem, the more limited resources and the more complex nature of the setting. The two groups most affected by Fe deficiency are young children and women of reproductive age. Infant diets in developing countries are low in iron, due to less use of industry prepared foods and much lower consumption of food from animal sources. Successful experiences in countries such as Chile and the United States have shown that it is feasible to reduce anemia levels in young children through the use of fortified infant food products and low cost weaning foods. In settings in which people are already using processed foods, the cost of improving the nutritional value of these foods is marginal compared with the significant benefits. However, costs and accessibility by the poorest are important concerns, and other options such as supplementation and efforts to improve complementary feeding also require attention. The high prevalence of iron and other micronutrient deficiencies due to poor diets and/or infections among women before and during pregnancy calls for strategies such as fortification and periodic supplementation. Experience to date suggests that fortification of staples (e.g., wheat flour) is a cost-effective and feasible strategy, but regulatory monitoring is required to demonstrate effectiveness and ensure quality. Supplementation is still required for all pregnant women, however, and challenges remain in finding ways to improve coverage and compliance. In summary, effectively combining and balancing the needs of program implementation, research and community involvement will help combat Fe deficiency. PMID- 11925491 TI - Eradication of iron deficiency anemia through food fortification: the role of the private sector. AB - Delivering iron fortified foods that provide meaningful levels of bioavailable iron without altering the accepted appearance and taste of the product presents multiple challenges. Issues relating to food technology, product formulation, acceptance and efficacy evaluation, marketing and quality control must all be addressed. Procter & Gamble Company has developed a unique technology that stabilizes iron in an aqueous system. Utilizing this technology, a fortified powder drink has been developed that is easy to distribute, store and use and that delivers 20-30% of the U. S. RDA for iron, as well as significant amounts of vitamin A, iodine, zinc and vitamin C in a single serving. Acceptance, bioavailability and effectiveness trials have all produced positive results. This type of fortified product can contribute to alleviating iron deficiency but requires scaling up, packaging, quality control and distribution through normal trade channels and public institutions to have a sustainable impact. To be effective, a well-planned communications campaign should also accompany any major iron fortification program. Eradication of iron deficiency anemia can be done but requires a holistic approach that addresses multiple barriers and leverages the untapped expertise and strength of the alliance between public and private sectors. PMID- 11925492 TI - Communication strategies to optimize commitments and investments in iron programming. AB - There is consensus that a communications component is crucial to the success of iron supplementation and fortification programs. However, in many instances, we have not applied what we know about successful advocacy and program communications to iron programs. Communication must play a larger and more central role in iron programs to overcome several common shortcomings and allow the use of new commitments and investments in iron programming to optimum advantage. One shortcoming is that iron program communication has been driven primarily by the supply side of the supply-demand continuum. That is, technical information has been given without thought for what people want to know or do. To overcome this, the communication component, which should be responsive to the consumer perspective, must be considered at program inception, not enlisted late in the program cycle as a remedy when interventions fail to reach their targets. Another shortcoming is the lack of program focus on behavior. Because the "technology" of iron, a supplement, or fortified or specific local food must be combined with appropriate consumer behavior, it is not enough to promote the technology. The appropriate use of technology must be ensured, and this requires precise and strategically crafted communications. A small number of projects from countries as diverse as Indonesia, Egypt, Nicaragua and Peru offer examples of successful communications efforts and strategies for adaptation by other countries. PMID- 11925493 TI - Policy and sustainability issues. AB - The need for combined and integrated strategies to address iron deficiency is widely recognized, utilizing targeted supplementation, as well as food based strategies including both fortified and nonfortified foods. The challenge is not so much knowing "what" to do as is understanding "how" to implement effective and sustainable interventions. Because the causes of iron and other micronutrient deficiencies are complex, including inadequate food intake, unsanitary conditions and inadequate health services, the solutions may also be complex, requiring multisectoral and interdisciplinary approaches. Top-down strategies are unlikely to be effective and sustainable. Rather, the beneficiaries of the program at the community level must be able to understand malnutrition in simple terms, to envisage potential solutions and to become "demanders" of services. The experience in Thailand provides an example of a country-wide, community-based and participatory approach utilizing facilitators and motivators at the local level to implement and sustain interventions. The experience in Thailand indicates the potential for developing effective and sustainable interventions to address iron deficiency and other micronutrient problems as part of a broad, community-based effort. PMID- 11925494 TI - Measuring performance: a strategy to improve programs. AB - This article stresses the importance of evaluation as a tool for improving the design and implementation of effective programs to reduce iron deficiency anemia and to advocate for their continued and increased support. Current concepts in program evaluation are applied to the specific issues relevant to iron programs. Evaluations should be designed to meet the needs of specific audiences (e.g., community members, program implementers, policymakers, donors and researchers) and to answer specific questions. Evaluations might answer questions about provision, coverage or impact. The choice of indicators for evaluating impact of iron programs is discussed and illustrated with recent examples. Evaluation design can be broadly categorized as monitoring, plausibility evaluations and probability evaluations. These designs differ in cost and also in the strength of evidence that they provide; however, each has appropriate uses. It is important to document program costs in the evaluation process as policymakers and donors weight impact against costs. To be useful, evaluation findings must be disseminated, usually to multiple audiences. This requires allocation of time and resources and attention to the needs of various stakeholders. PMID- 11925495 TI - Iron fortification of foods: overcoming technical and practical barriers. AB - Iron fortification of food is a methodology utilized worldwide to address iron deficiency. A critical problem in some food fortification programs is the lack of bioavailability of iron compounds. This article reviews presentations on iron fortification programs in the Americas and the technical and practical barriers faced by the programs. Effective programmatic strategies will incorporate systematic consideration of sound program management, ensure consumption of fortified foods, and promote advocacy and supportive legislation. However, these factors are often overlooked or are not addressed coherently. Key components to consider in implementation of iron fortification programs include: utilization of foods and condiments containing fortificants that are bioavailable; program development and its implementation coordinated with targeted communications; utilizing resources from public/private sector partnerships; and operational research on how to overcome practical barriers for successful implementation of fortification. PMID- 11925496 TI - Iron supplementation: overcoming technical and practical barriers. AB - Iron supplementation is probably the best available option to effectively address iron deficiency in pregnant women and young children because it can be targeted specifically to these high-risk groups. However, technical and practical barriers exist: limited information on the effectiveness of supplementation interventions, side effects that affect compliance, and supply/distribution constraints. An innovative approach to addressing these constraints is the use of sprinkles of powdered, microencapsulated ferrous fumarate that can be added directly to any semi-liquid food without changing their taste or consistency. This technique has been tested in initial trials in Ghana and found to be as effective as iron drops. Another approach to improve the effectiveness of iron interventions is through information, education and communication (IEC) programs. These interventions can help modify consumer behavior in some cases, but in some countries, geographic location, variations in language and population size can make the cost of IEC programs very high. IEC strategies in Indonesia aimed at increasing demand for iron supplements by systematic dissemination of specific messages, improving the quality and variety of tablets, increasing the availability and access to supplements by engaging the commercial sector, enrolling traditional birth attendants and other community volunteers in selling supplements. Key issues to be addressed include clarifying optimal starting points and duration of supplementation interventions--based on individual status or population prevalence, defining hemoglobin and ferritin cutoffs at which treatment should be instigated and evaluating the effectiveness of intermittent supplementation with multiple micronutrients. PMID- 11925497 TI - Iron fortification: country level experiences and lessons learned. AB - Iron fortification has been used to enhance iron intake in many developed countries for more than 50 years, but only in the last decade has this strategy been applied on a large scale to other parts of the world. Iron fortification of rice is being instituted in the Philippines. Initially, the rice will be produced in government-controlled rice mills and sold at low cost mainly to low income families. Efforts to improve the technology (using coating or extrusion techniques) are currently underway to reduce cost and minimize losses during storage and washing. Effectiveness and feasibility studies are required to test the new technologies and processing/distribution systems. In Venezuela in 1993, the government instituted a mandatory program of iron fortification to enrich precooked corn flour followed by the voluntary fortification of wheat flour. Surveys in school children subsequently showed a sharp drop in iron deficiency. Fortification of fish sauce in Vietnam has shown promising initial results in reducing anemia among anemic, nonpregnant female factory workers. Iron-fortified soy sauce has been shown to reduce anemia in initial studies in children in China, and a large-scale efficacy trial is now underway. These examples indicate that iron fortification of staple foods and condiments holds great promise for the prevention of iron deficiency. PMID- 11925498 TI - Iron supplementation: country level experiences and lessons learned. AB - Iron supplementation is a commonly used strategy to meet the increased requirements of at-risk groups, such as women of childbearing age, especially during pregnancy. Other at-risk groups for which iron supplementation may be appropriate include infants, young children, adolescents and the elderly. There is a need to consider iron supplementation as part of a comprehensive strategy for the prevention of iron deficiency, and not just as a treatment for anemia that is stopped as soon as clinical improvement is noted. Experience in developing countries indicates that often the poorest women with the most deficient intakes are the least likely to receive iron supplements during pregnancy. Providing supplements to women during antenatal care visits is useful but often inadequate, so other delivery channels must also be explored, including private sector markets and community networks. Communication efforts must be expanded to increase understanding of the importance of taking supplements and to address any fears or misconceptions relating to supplementation. Overall, we must increase the capacity of individuals and communities to define, analyze and act to address their own health needs. PMID- 11925499 TI - Prevention and control of anemia: Thailand experiences. AB - Thailand has addressed nutrition in national development policy since the mid 1970s, including efforts to reduce iron deficiency anemia. Nutritional improvement has been implemented as an integral part of primary health care and community development extending beyond government services to include community participation. Utilization of village health volunteers has been a crucial feature of the program. Available data indicate that anemia rates have declined among pregnant women and preschool children, although there has been no formal evaluation of the program effect. Universal iron supplementation has been the major strategy for pregnant women, using village health volunteers to encourage continuation of the antenatal care schedule and encouraging a preventive approach by health service providers. Program obstacles have included lack of access to iron tablets by some populations and lack of understanding of the importance of anemia. Women's compliance was complicated by fear of having a large fetus, forgetfulness and side effects. Weekly iron supplementation of school children was piloted in 2000, and is now being extended. Other strategies utilized to address iron deficiency include food fortification, dietary improvement and complementary public health measures. Program monitoring and evaluation require strengthening to assess the effectiveness of intervention strategies and provide proper data for decision-making. PMID- 11925500 TI - Anemia prevention and control in four central Asian republics and Kazakhstan. AB - Kazakhstan and the central Asian republics of Uzbekistan, the Kyrgyz Republic, Tajikistan and Turkmenistan have developed anemia prevention and control (APC) policies based on multiple interventions, including education and promotion, oral supplementation of high risk groups and fortification of wheat flour with iron and other micronutrients. These national strategies are aimed at reducing the prevalence of anemia and iron deficiency among young children and women of child bearing age. Strategy development has been assisted by funding and technical assistance from the United Nations Children's Fund (UNICEF) with additional technical support from the International Nutrition Foundation, the United Nations University and various national institutions. These countries have been among the most advanced in adopting national strategies that include multiple interventions in an overall package, and national interest in APC remains high. However, reviews of APC activities conducted in 2001 suggests the need for modification and enhancement of current efforts and for a shift to national-level actions if these countries are to progress toward current and future goals. Increased commitment and determination, by both national groups and international organizations, are required to achieve and sustain improvement in micronutrient nutrition. PMID- 11925501 TI - Panel discussion: regional action priorities. AB - Action priorities for reducing iron deficiency vary across regions. Some regions have limited experience with national programs, such as in the republics of Central Asia, where the focus since independence has been on assessment, advocacy and scaling up of interventions. In more developed regions the priorities are improving established programs and addressing the needs of the remaining subpopulations with high rates of iron deficiency. The etiology of anemia, whether multi-factorial as in Sub-Saharan Africa or mostly due to iron deficiency as in the Middle East and North Africa, is an important factor shaping the choice of strategies. Some interventions are particularly feasible in some settings and are being promoted aggressively, such as flour fortification in Latin America and the Middle East and North Africa, or the fortification of condiments in Asia. The quality of the health infrastructure and resources in general also determine the mix and scale of interventions implemented. A lesson for all regions is that countries in North America and Europe, where diets are varied and abundant and infections nearly absent, generally support multiple strategies that include nutrition education, supplementation and fortification of a variety of staples consumed by toddlers and the general population. Monitoring and evaluation efforts have proven necessary for improving quality and for advocacy. PMID- 11925502 TI - Summary and recommendations. AB - Iron deficiency affects the well being of > 1 billion people worldwide, with a range of adverse health and social consequences, yet efforts to address the problem have had only limited success. To better assess this issue and define more effective strategies, an international conference was convened in Atlanta, GA, in May 2001. Key policy issues discussed included setting a global goal for prevention and control of iron deficiency, building partnerships, and identifying ways to mobilize financial and human resources. The strengths and limitations of specific intervention strategies were discussed including iron fortification of staple foods and condiments, supplementation, dietary diversification, application of biotechnology to increase micronutrient content of staple foods, and public health measures such as infection control and provision of health care services. The importance of utilizing multiple and integrated strategies was emphasized. Representatives from several countries reported successful intervention programs as well as promising results in implementing new strategies. The critical role of effective communications was also emphasized, both to increase awareness of the impact of iron deficiency and to advocate for policy changes and resources to address the problem. The conference concluded with specific recommendations for action. PMID- 11925503 TI - Prevention and control of iron deficiency: priorities and action steps. AB - The strengthening of global efforts to prevent and control iron deficiency requires priority setting and action steps in several key areas, including research, partnership formation, policy setting and the integration of intervention strategies. Research priorities include the development of improved assessment tools, evaluation of fortification strategies, improvement in interventions for infants, evaluation of combined intervention strategies to address multiple micronutrients and development and testing of interventions using genetically engineered foods with improved nutritional qualities. Policy priorities include the expansion of partnerships, balancing of advocacy, research and program implementation, and improved communications. Priorities in partnership formation include building strong linkages between public sector efforts and the food industry to enhance training, technical expertise and advocacy. Strategies to address iron deficiency should seek to integrate efforts in food fortification, supplementation, dietary improvement and complementary public health measures. PMID- 11925504 TI - Oligosaccharides of milk and colostrum in non-human mammals. AB - Mammalian milk or colostrum usually contains, in addition to lactose, a variety of neutral and acidic oligosaccharides. Although the oligosaccharides of human milk have been reviewed in several recent publications, those of non-human mammals have received much less attention. This paper reviews the chemical structures and the variety of milk oligosaccharides in species other than humans, including placental mammals (e.g. primates, domestic herbivores, bears and other carnivores, the rat and the elephant) as well as monotremes (platypus and echidna) and marsupials (e.g. wallaby). The gastrointestinal digestion and absorption and the possible biological functions of these oligosaccharides are also discussed. PMID- 11925505 TI - N- and O-linked oligosaccharides of allergenic glycoproteins. AB - Cross-linking of cell-bound IgE on mast cells or basophils by polyvalent antigens causes the release of histamine and other mediators of the allergic response which then lead to the development of allergic symptoms. In this event not only peptide epitopes, but also carbohydrates can act as cross-linking elements. Since peptide epitopes of allergens are subject of most published studies, this review is focused on glycosidic epitopes. The current knowledge of the structures and possible epitopes of oligosaccharides linked to allergenic glycoproteins is briefly reviewed, showing that complex plant N-glycans containing alpha1,3 fucose and beta1,2 xylose are most frequently involved in the structures of IgE epitopes. In own studies a prevalence of up to 29% anti-glycan IgE was determined among pollen-allergic patients. The clinical relevance of these carbohydrate specific IgE antibodies is still a matter of controversial discussions. PMID- 11925506 TI - Characterization of the sugar-binding specificity of the toxic lectins isolated from Abrus pulchellus seeds. AB - The sugar-binding specificity of the toxic lectins from Abrus pulchellus seeds was investigated by combination of affinity chromatography of glycopeptides and oligosaccharides of well-defined structures on a lectin-Sepharose column and measurement of the kinetic interactions in real time towards immobilized glycoproteins. The lectins showed strong affinity for a series of bi- and triantennary N-acetyllactosamine type glycans. The related asialo oligosaccharides interact more strongly with the lectins. The best recognized structures were asialo-glycopeptides from fetuin. Accordingly, the kinetic interaction with immobilized asialofetuin was by far the most pronounced. Human and bovine lactotransferrins and human serotransferrin interacted to a lesser extent. The interaction with asialofetuin was inhibited by galactose in a dose dependent manner. Lactose, N-acetyllactosamine and lacto-N-biose exhibited similar degree of inhibition while N-acetylgalactosamine was a poor inhibitor. These results suggested that the carbohydrate-binding site of the Abrus pulchellus lectins was specific for galactose and possess a remarkable affinity for the sequences lactose [beta-D-Gal-(1-->4)-D-Glc], N-acetyllactosamine [beta-D Gal-(1-->4)-D-GlcNAc] and lacto-N-biose [beta-D-Gal-(1-->3)-D-GlcNAc]. PMID- 11925507 TI - Requirement of chondroitin sulfate/dermatan sulfate recognition in midkine dependent migration of macrophages. AB - Midkine (MK) is a heparin-binding growth factor that promotes cell migration, cell growth and cell survival. The promotion of migration of inflammatory cells, especially macrophages, by MK is involved in formation of a vascular abnormality, i.e. neointima formation. MK-induced migration of peritoneal exudate macrophages was inhibited by heparin, chondroitin sulfate E and dermatan sulfate, but not by chondroitin sulfate D or chondroitin 6-sulfate. Digestion of macrophages with chondroitinase ABC as well as chondroitinase B decreased the migratory activity. However, heparitinase digestion showed only slight effects. These results indicated that a chondroitin sulfate, i.e. an E-type oversulfated structure with dermatan sulfate domain, is involved in MK-induced migration of macrophages. Although a chondroitin sulfate proteoglycan, receptor-type protein tyrosine phosphatase zeta (PTP zeta), participates in MK-induced migration of neurons and osteoblasts, PTP zeta was not detected in macrophages. The MK-induced migration was inhibited by PP1, wortomanin, PD 98059 and vanadate, indicating that the downstream signaling system, which includes Src, PI3 kinase and ERK as important components, is shared with other MK signaling systems in which PTP zeta is involved. PMID- 11925510 TI - The efficacy of ECT and "treatment resistance". PMID- 11925508 TI - Different behavior of ghost-linked acidic and neutral sialidases during human erythrocyte ageing. AB - Acidic and neutral sialidases (pH optimum 4.7 and 7.2, respectively) were assayed on human circulating erythrocytes during ageing. The assays were performed on intact erythrocytes and resealed erythrocyte ghost membranes. From young to senescent erythrocytes the acidic sialidase featured a 2.7-fold and 2.5-fold decrease in specific activity when measured on intact cells or resealed ghost membranes, whereas the neutral sialidase a 5-fold and 7-fold increase, respectively. The Ca2+-loading procedure was employed to mimic the vesiculation process occurring during erythrocyte ageing. Under these conditions the released vesicles displayed an elevated content of acidic sialidase, almost completely linked through a glycan phosphoinositide (GPI) anchor but no neutral sialidase activity, that was completely retained by remnant erythrocytes together with almost all the starting content of sialoglycoconjugates. The loss with vesiculation of acidic sialidase with a concomitant relative increase of neutral sialidase was more marked in young than senescent erythrocytes. The data presented suggest that during ageing erythrocytes loose acidic sialidase, and get enriched in the neutral enzyme, the vesiculation process, possibly involving GPI anchors-rich membrane microdomains, being likely responsible for these changes. The enhanced neutral sialidase activity might account for the sialic acid loss occurring during erythrocyte ageing. PMID- 11925509 TI - The effect of alpha2,6-linked sialic acid on anti-IgM antibody-induced apoptosis in Ramos cells. AB - Apoptosis in B cells is induced through the B cell antigen receptor (BCR) and affects the sialic acid recognition molecules on B cells. We investigated the effects of alpha(1)-acid glycoprotein (AGP), which mainly contains alpha2,6 linked sialic acid, on anti-IgM antibody (Ab)-induced apoptosis in Ramos cells, which are derived from Burkitt's lymphoma. When Ramos cells were incubated with anti-IgM-Ab in plates coated with AGP, neuraminidase-digested AGP (asAGP) or alpha2,3-sialylated AGP (2,3AGP), apoptosis was suppressed only in those coated with AGP. We also studied the effects of CD22, which is expressed on the surface of mature B cells and binds to sugar chains containing alpha2,6-linked sialic acid, with anti-CD22 monoclonal antibody (mAb). Anti-CD22mAb enhanced anti-IgM Ab induced apoptosis in Ramos cells. These contradictory results suggested that the recognition molecules for alpha2,6-linked sialic acid on AGP, which inhibits B cell apoptosis, is distinct from CD22, or that different binding domains of CD22 between alpha2,6-linked sialic acid and anti-CD22 mAb exert opposite functions of suppression or enhancement to anti-IgM Ab-induced B cells. PMID- 11925511 TI - Stimulus titration and ECT dosing. AB - The seizure threshold to electroconvulsive therapy (ECT) is defined entirely by the duration of the induced seizure, is multidetermined, and varies enormously with a wide variety of patient and treatment factors, including especially the parameters of the ECT stimulus. No consistent relationship has ever been detected between the clinical antidepressant response to ECT and either the threshold or the duration of the induced seizure. The stimulus titration method for determining the seizure threshold (titration-threshold dosing) was the central research tool used to reverse years of dogma by proving that the induced seizure of ECT is not alone sufficient to explain the therapeutic properties of ECT, and that the interaction between dosage and treatment electrode placement is critical in this regard. In clinical use, however, titration-threshold dosing has proven less than fully effective in optimizing the stimulus dose for ECT-better results are consistently obtained with age-based or fixed high-dose methods. The lack of a direct correlation between either seizure threshold or duration and clinical ECT response is an irremediable flaw of the titration-threshold method as clinically applied. New approaches are now called for in which ECT stimulus dosage is set and adjusted ("titrated") according to the clinical antidepressant response of the patient or to measurable correlates thereof: maximum sustained electroencephalogram (EEG) ictal power, EEG postictal suppression, induced interictal EEG delta activity, peak heart rate, maximum sustained EEG coherence, and postictal EEG coherence reduction, all of which have been found by various investigators to be related to the clinical antidepressant response to ECT. PMID- 11925516 TI - Safety, efficacy, and effects on glycemic control of electroconvulsive therapy in insulin-requiring type 2 diabetic patients. AB - OBJECTIVE: To determine the safety, efficacy, and effects on glycemic control of electroconvulsive therapy (ECT) in insulin-requiring type 2 diabetic patients. METHODS: Chart review was conducted of 19 patients with insulin-requiring type 2 diabetes mellitus who underwent ECT at Mayo Clinic (Rochester, Minnesota) between 1993 and 2000. None of the patients received oral hypoglycemic drugs. Daily fasting blood glucose concentrations and daily insulin requirements were determined. For each patient, the daily insulin requirements before the first ECT treatment were compared with insulin requirements after the last ECT treatment. In addition, the insulin requirements 1 day before and 1 day after each ECT treatment (n = 143) were compared. Charts were reviewed for activity, appetite, and weight changes. RESULTS: The average change in daily insulin requirements 1 day before the initiation of ECT compared with 1 day after the completion of ECT was not statistically significant. Changes in dietary or activity levels correlated with fluctuations in insulin requirements in selected individuals. There were no significant changes in acute glycemic control associated with ECT. CONCLUSIONS: Our findings refute previous reports of dangerous hyperglycemia or improved glycemic control due to ECT in insulin-requiring type 2 diabetes, and suggest that ECT is safe and efficacious in these patients. PMID- 11925517 TI - A switch from propofol to etomidate during an ECT course increases EEG and motor seizure duration. AB - INTRODUCTION: Of 58 patients treated at our electroconvulsive therapy (ECT) unit early in the year 2000, 12 patients under propofol did not achieve a seizure duration of >30 s [electroencephalogram (EEG)] with a maximum stimulation charge of 504.0 mC (100%). METHOD: A switch from propofol to etomidate was therefore undertaken in these patients at the next treatment to achieve longer seizure duration. RESULTS: In 11 of the 12 patients, a remarkable increase in seizure duration was recorded after the change of anesthetic. The mean seizure duration increased from 18.6 to 43.4 s and remained at that level for the following ECT session. The increase was highly significant (t11 = 3.772, p < 0.001). The mean motor seizure also changed from 11.6 to 27.5 s (t11 = 5.560, p < 0.003) and remained there for the next treatment. DISCUSSION: Our data show that the switch more than doubles EEG seizure duration and suggest that etomidate can be used instead of methohexital as an alternative in patients with short seizure duration. It is also a potential option to avoid the pain frequently associated with the injection of propofol. PMID- 11925519 TI - Effects of stimulus parameters on seizure duration and ECS-induced retrograde amnesia. AB - A selected electroconvulsive therapy stimulus dose can result from different combinations of pulse amplitude, pulse width, pulse frequency, and stimulus duration; however, the roles of these stimulus variables in the effects of the overall stimulus dose are not clearly understood. A series of studies on Wistar rats was therefore conducted to assess the effects of different stimulus compositions at constant stimulus charge. In the first two studies, two differently composed 60 mC charge unidirectional electroconvulsive shock (ECS) stimuli were administered once daily for 3 days, while in the third study three differently composed 120 mC charge unidirectional ECS stimuli were administered on a single occasion. At constant charge, a markedly longer stimulus duration was associated with ear burns and an approximately 12.5% longer seizure duration. At constant charge, however, different stimulus compositions were not associated with different degrees of impairment in two passive avoidance models of ECS induced retrograde amnesia. These preliminary findings have implications for the choice of stimulus settings during dose titration and dose selection procedures. While the findings encourage further investigation, it appears that variations in stimulus duration are biologically significant, but minor variations in other stimulus elements may not influence the effects of electrically induced seizures very much. PMID- 11925518 TI - Right unilateral and bifrontal electroconvulsive therapy in the treatment of depression: a preliminary study. AB - The short-term outcome of electroconvulsive therapy (ECT) was studied in 24 patients with a current major depressive episode (DSM-IV). Patients were randomized to high dose (400% above the seizure threshold) right unilateral (RUL) ECT, to moderate dose (150% above seizure threshold) RUL ECT, and to low dose (just above seizure threshold) bifrontal (BF) ECT. Primary outcome measures included number of treatments, Hamilton Depression Rating Scale score, and Mini Mental State Examination score. High dose RUL ECT was associated with a significantly faster response to treatment than low dose BF ECT. Moreover, there was a tendency to a higher response rate with high dose RUL ECT compared with either moderate dose RUL ECT or BF ECT. PMID- 11925520 TI - ECT: use in individuals with mental retardation. AB - Electroconvulsive therapy (ECT) is an effective treatment of a variety of serious psychiatric and neurologic disorders. There are infrequent case reports of its use in individuals with mental retardation (MR). We describe 10 patients with MR and complex comorbid psychiatric disorder treated with ECT. Seven patients had an excellent response to treatment. Side effects of treatment were minimal and transitory. PMID- 11925521 TI - Medical student knowledge and attitudes regarding ECT prior to and after viewing ECT scenes from movies. AB - We surveyed samples of medical students in the United Kingdom (U.K.) and Australia, prior to their psychiatry placement, to ascertain views about electroconvulsive therapy (ECT) and the effect on those views of watching ECT scenes in movies. A 26-item questionnaire was constructed by the authors and administered to the students. At set times during the questionnaire, students were asked to view five movie clips showing, or making reference to, ECT. The clips were from Return to Oz, The Hudsucker Proxy, Ordinary People, One Flew Over the Cuckoo's Nest, and Beverly Hillbillies. Ninety-four students participated in the study. Levels of knowledge about the indications, side effects, and mode of administration were poor, and attitudes were generally negative. Viewing the ECT scenes influenced attitudes toward the treatment; after viewing, one-third of the students decreased their support for ECT, and the proportion of students who would dissuade a family member or friend from having ECT rose from less than 10% to almost 25%. PMID- 11925522 TI - ECT in poetry. AB - We sought to identify poems about Electroconvulsive therapy (ECT) and describe their major themes, with a view to using some of those poems in clinical situations-for persons receiving ECT and their families-as well as for teaching purposes. Several poems were identified, eight almost exclusively about ECT and a number of other works containing references to the treatment. The poets, some of whom have won distinguished prizes, came from the United States, U.K., Australia, and Estonia. The poems included very positive accounts of ECT and more denigrating pieces. Some of the themes covered by the poems were the desperate plight of persons prior to ECT, the potentially striking beneficial effects of the treatment, fears about the use of electricity, ECT procedure, and memory impairment, and the stigma associated with the treatment. PMID- 11925523 TI - ECT for the treatment of psychiatric symptoms in Basedow's disease. AB - To treat psychiatric symptoms associated with hyperthyroidism, we administered electroconvulsive therapy (ECT) to a patient with Basedow's disease. After unsuccessful treatment with an oral antithyroid drug, neuroleptics, and mood stabilizer, the patient recovered completely after three ECTs and did not relapse in the 40-day follow-up period. Although hyperthyroidism has been considered to induce psychiatric symptoms by enhancement of the sensitivity and turnover in catecholaminergic neurotransmission, we suggest another possible mechanism, involving translational and activational regulation of functional proteins in the brain. PMID- 11925524 TI - The practice of electroconvulsive therapy: recommendations for treatment, training, and privileging (second edition). PMID- 11925525 TI - ECT for depression with amnesia. PMID- 11925527 TI - The "brush" controversy. PMID- 11925526 TI - Sequence and spatial expression of zebrafish (Danio rerio) alphaA-crystallin. AB - PURPOSE: To determine the nucleotide sequence, amino acid sequence and tissue specificity of zebrafish alphaA-crystallin. METHODS: RACE, both 3' and 5', was used to clone the zebrafish alphaA-crystallin gene. The peptide sequence of the encoded protein was deduced and compared to cavefish, shark, amphibian, bird and human orthologues using the CLUSTAL W algorithm. alphaA-crystallin transcript was evaluated in brain, heart, lens, liver, skeletal muscle/skin, and spleen by semi quantitative RT-PCR. RESULTS: The 173 amino acid sequence of zebrafish alphaA crystallin was determined to be 73% and 86% similar to its human and cavefish orthologues, respectively. We detected high expression of zebrafish alphaA crystallin in the lens and very low expression in liver and spleen. CONCLUSIONS: Few amino acids identified as being functionally important to chaperone function differ between zebrafish and mammalian alphaA-crystallin. The expression of alphaA-crystallin is mainly confined to the lens in both taxa. These data suggest that zebrafish alphaA-crystallin plays a physiologically limited role outside of the zebrafish lens, similar to its mammalian orthologues. PMID- 11925528 TI - Oral candidiasis in human immunodeficiency virus-infected women. PMID- 11925529 TI - Bilateral parotid swelling: a review. PMID- 11925530 TI - Demographics and treatment options for orbital roof fractures. AB - OBJECTIVE: The purpose of this article was to review the frequency, germane anatomy, management modalities, and complications associated with the treatment of orbital roof fractures in the pediatric and the adult population. STUDY DESIGN: A review of the past 30 years of the English-language maxillofacial surgical literature was undertaken. Important concepts were coupled with the authors' experience to provide a synopsis of contemporary thought on this topic. RESULTS: More than 235 articles in the oral and maxillofacial, plastic and reconstructive, otolaryngology-head and neck, ophthalmologic, oculoplastic, neurologic, and pediatric surgical literature were reviewed and assessed. From this group, 50 articles were found to contain useful information. CONCLUSIONS: It has been estimated that 1% to 9% of all facial fractures involve the orbital roof. The typical adult with an orbital roof fracture is a man (89%-93%) who has been involved in a high-energy impact and who has sustained concomitant multisystem injuries (57%-77%). Orbital roof fractures most commonly coexist with other craniofacial injuries. In contrast, in pediatric patients with an orbital roof injury, we see nearly equal sex distribution; the typical patient in this case has a frontobasal fracture that is minimally displaced or nondisplaced (53% 93%) and has sustained concomitant multisystem injuries. The pediatric patient is usually managed by means of observation alone (53%-86%). For the adult patient, a subcranial approach to the orbital roof by means of a bitemporal flap or superior blepharoplasty incision offers wide access with minimal morbidity. Currently available titanium microscrew and miniscrew and mesh systems offer a near-ideal modality for orbital roof reconstruction. The coexisting neurocranial, frontal sinus, and supraorbital rim fractures take priority over the management of orbital roof fractures. Complications associated with orbital roof injuries can be categorized as those attributed to the following: concomitant injury, surgical access, postreconstruction volume discrepancy, muscle entrapment, hemorrhage, and/or infection. PMID- 11925532 TI - The expression of vascular endothelial growth factor in synovial tissues in patients with internal derangement of the temporomandibular joint. AB - OBJECTIVE: The aim of this study was to elucidate the expression and localization of vascular endothelial growth factor (VEGF) in synovial tissue taken from the temporomandibular joint (TMJ) with internal derangement (ID) and discuss the role of VEGF in the pathogenesis of ID. STUDY DESIGN: Through the use of an immunohistochemical technique, 39 TMJs in 37 patients were examined. As controls, synovial tissue specimens from 6 joints in 6 patients with habitual dislocation were also examined. RESULTS: In the synovial tissue from 35 of the patients with ID, expression of VEGF was observed in the synovial lining cells, in the endothelial cells of the blood vessels, and in the fibroblasts. In contrast, expression of VEGF was found in the TMJ tissue from only 2 of the controls. The percentage of VEGF-positive cells in the ID specimens was significantly higher than that in the habitual dislocation specimens (P < .02), and the expression of VEGF significantly correlated with the arthroscopic synovitis score (P = .004). CONCLUSION: These results suggest that the expression of VEGF is upregulated and involved in the development of inflammatory changes in synovial tissues in TMJs with ID. PMID- 11925531 TI - Management of dental extractions in patients with bleeding disorders. AB - OBJECTIVE: This study evaluated the effectiveness of a protocol to prevent bleeding after dental extraction in patients with hemophilia, von Willebrand's disease (VWD), or platelet disorders. STUDY DESIGN: Replacement therapy was used in cases involving general anesthesia, and nerve trunk infiltration was used in patients with severe bleeding disorders (severe-to-moderate hemophilia or type 2 3 VWD). Desmopressin was used in good responders with mild hemophilia A, type 1 VWD, and platelet disorders. Local hemostatic measures and antifibrinolytic treatment were used systematically. RESULTS: Ninety-three patients underwent 103 dental extractions; 2 of these patients had secondary bleeding requiring surgical hemostasis. CONCLUSION: The indication for replacement therapy depended on the type of anesthesia that was used. Coagulation factor concentrates or desmopressin were necessary to avoid upper airway hematoma with general anesthesia or nerve trunk infiltration. With local anesthesia, substitutive treatment was indicated in patients with severe-to-moderate hemophilia and type 2-3 VWD. Inexpensive desmopressin was effective in those who responded well. Local hemostatic measures and antifibrinolytic treatment were performed systematically. PMID- 11925533 TI - Acute myelogenous leukemia: advances and limitations of treatment. AB - Acute myelogenous leukemia (AML) describes a group of related hematologic malignancies that are being approached therapeutically from several perspectives. Conventional chemotherapeutic agents, such as anthracyclines and cytosine arabinoside (Ara-C), are useful in treating AML but now appear to have reached their maximum potential. Newer therapeutic approaches to AML have recently focused on immune-based therapy through monoclonal antibodies that target and destroy malignant cells via specific cell receptors. One such agent is gemtuzumab (CMA-676), an agent that targets the CD33 antigen on malignant myeloid cells. Initial studies have shown significant anticancer activity. We will discuss traditional and newer therapeutic approaches to AML and review the role of monoclonal antibody based therapies for patients with AML. A case of a 30-year old man with refractory AML who was treated with gemtuzumab will be mentioned, highlighting potential applications and possible limitations to this novel therapy. Despite the effective reduction in the number of malignant cells in bone marrow, gemtuzumab ineffectively treated extramedullary leukemic gingival infiltrate. Regardless of limitations, monoclonal-based therapy offers an exciting and potentially safer adjunctive therapy for patients with AML. PMID- 11925534 TI - Treatment of severe chronic oral erosive lesions with clobetasol propionate in aqueous solution. AB - OBJECTIVE: We sought to analyze the results of topical treatment with a mouthwash of 0.05% clobetasol in aqueous solution in 30 patients with severe oral erosive lesions. STUDY DESIGN: Over a 48-week period, we evaluated the evolution of pain, ulcerations, atrophy, and interference of the disease in the patient's daily life, classifying the response as complete (100% remission/recovery), excellent (75%), good (50%), poor (<50%), or failed. RESULTS: The pain and ulceration totally disappeared in 93.3% of cases and 90% reported a full recovery in their daily life activities. Atrophy response was complete in 28.5%, excellent in 60.7%, and good in 3.5%. Two patients showed no response to the treatment. Five patients suffered mild adverse effects (moon face and hirsutism) between week 4 and week 6 of treatment, which were speedily reversed by reducing the frequency of mouthwash. CONCLUSIONS: Clobetasol mouthwash is a safe and efficacious option for the treatment of severe oral erosive lesions. PMID- 11925535 TI - A clinical trial of the anticaries efficacy of casein derivatives complexed with calcium phosphate in patients with salivary gland dysfunction. AB - OBJECTIVE: The purpose of this study was to compare the caries preventive efficacy of a mouthrinse solution containing casein derivatives coupled with calcium phosphate (CD-CP) with that of a 0.05% sodium fluoride mouthrinse among individuals with dry mouth. STUDY DESIGN: A randomized control trial design was used. Participants included individuals who had had radiotherapy for head and neck cancer (n = 82) and others with Sjogren's syndrome (n = 56). Baseline data collection was followed by reexamination 12 months later. Posterior bite-wing radiographs were taken on both occasions. RESULTS: A total of 124 participants, 61 (49.2%) in the sodium fluoride group and 63 (50.8%) in the CD-CP group, completed the 12-month examination. The baseline characteristics of the 2 groups did not differ. Coronal caries incidence was higher in the sodium fluoride group than in the CD-CP group (34.4% and 27%, respectively), but the difference was not statistically significant. Similarly, the small difference in coronal caries increment between the 2 groups was not statistically significant (0.4 and 0.3 surfaces, respectively). There was insufficient root surface caries experience between the 2 groups observed for differences to be examined. Proportionately more of the CD-CP group lost 1 or more teeth, and the mean number of tooth loss was higher. The participants with the highest incidence and increment were those with Sjogren's syndrome in the CD-CP group. Some of that difference was accounted for by differences in baseline caries status. CONCLUSION: It appears that CD-CP preparations hold promise as caries preventive agents for individuals with dry mouth, although confirmation of this study's findings in other settings is warranted before a definitive conclusion can be reached. PMID- 11925536 TI - Klippel-Trenaunay syndrome (angio osteohypertrophy syndrome): a report of 3 cases. AB - Klippel-Trenaunay syndrome was first reported in 1900 by Klippel and Trenaunay. The syndrome is characterized by the triad of vascular nevi, venous varicosity, and hyperplasia of soft tissue-and possibly bone-in the affected area. The original description was made with regard to the extremities. The syndrome is diagnosed on the basis of 2 or 3 symptoms. The 3 patients reported here presented with hemangioma and hypertrophy of the jawbone and concomitant malocclusion. There were clinically no obvious varicose veins. It appears that gravity plays an important role in the venous drainage from the head and neck region, and this could be the reason for the rarity of varicose veins in the head and neck region when compared with lower extremities. In addition, 2 of the 3 cases showed exophytic pedunculated growth arising from the gingiva, which has not been reported in any previous case reports. The following report presents the clinical features in the orofacial region and highlights the clinical significance of this syndrome. PMID- 11925538 TI - Calcifying epithelial odontogenic tumor showing microscopic features of potential malignant behavior. AB - Calcifying epithelial odontogenic tumor (CEOT) is a rare benign, but locally aggressive, odontogenic tumor, and only 2 cases of malignant CEOT are reported in the literature. We describe a case of an atypical CEOT that penetrates the blood vessels, invades bone, and perforates the cortical plates of the mandible. On histologic examination, it shows marked pleomorphism and numerous mitotic figures, including a tripolar mitotic figure. Proliferating activity was found to be 5 times higher than typical CEOTs as demonstrated by the proliferating index, Ki-67, and analyzed by a computerized image analysis system. The Ki-67 labeling index of this case was also compared to various previously reported benign and malignant neoplasms. Although there is no clinical finding of metastasis, we believe this neoplasm has malignant potential on the basis of the histologic features of vascular invasion, significant mitotic activity, atypical mitotic figures, and an increased proliferating index. PMID- 11925537 TI - Candidal carriage in the oral cavity of human immunodeficiency virus-infected subjects. AB - OBJECTIVES: We sought to assess asymptomatic oral carriage of Candida species and relative density in human immunodeficiency virus-infected (HIV+) subjects, as well as to determine whether yeast carriage is associated with CD4+ cell count, HIV-1 RNA quantity, gender, route of HIV transmission, antiretroviral therapy, or smoking. STUDY DESIGN: A cross-sectional analysis of oral rinses from HIV+ and healthy subjects was undertaken. Oral candidal carriage and relative species were investigated in 42 HIV+ and 41 healthy individuals, all of whom were seen at the Palermo University Department of Oral Sciences, Italy. Data were managed and analyzed by means of the computer software package StatView 5.0.1. RESULTS: Carriage rate was 61.9% and 29.3% (P =.003) among HIV+ and healthy subjects, respectively. Similarly, density carriage in the HIV+ group was found to be significantly higher than in the control group (P = .0002). Among HIV+ subjects, Candida carriage was significantly associated with smoking, whereas density was not. Oral candidal carriage and relative density were not significantly associated with the other 4 parameters evaluated. Candida albicans was the most frequently recovered species. CONCLUSIONS: Asymptomatic candidal carriage and relative density were found to be significantly higher in the oral cavity of HIV+ subjects-but not associated with CD4+ counts or HIV-1 RNA quantities. PMID- 11925539 TI - Psammomatoid and trabecular juvenile ossifying fibroma of the craniofacial skeleton: two distinct clinicopathologic entities. AB - The term juvenile ossifying fibroma is used in the literature in naming 2 microscopically distinct fibro-osseous lesions of the craniofacial skeleton. One is characterized by small uniform spherical ossicles resembling psammoma bodies (psammomatoid juvenile ossifying fibroma). The other is distinguished by trabeculae of fibrillary osteoid and woven bone (trabecular juvenile ossifying fibroma). Three new cases of each type are reported, and the literature is extensively reviewed for published cases of these 2 entities.Psammomatoid juvenile ossifying fibroma is reported more commonly than trabecular juvenile ossifying fibroma. It affects patients from a wider age range (3 months to 72 years) and an older mean age range (16-33 years) as compared with 2 to 12 years and 8(1/2) to 12 years, respectively, for trabecular juvenile ossifying fibroma. In both types there is a slight male predominance and the lesions are unencapsulated and tend to infiltrate adjacent bone. A significant difference between the 2 tumors is their site of occurrence. Although psammomatoid juvenile ossifying fibroma occurs predominantly in the sinonasal and orbital bones, trabecular juvenile ossifying fibroma predominantly affects the jaws. Aggressive growth occurs in some-but not all-cases of both types. Such behavior may be related to younger patient age and the concurrent development of aneurysmal bone cysts, which is seen more frequently in psammomatoid juvenile ossifying fibroma. This study demonstrates that not only histologic but also demographic and clinical differences between psammomatoid juvenile ossifying fibroma and trabecular juvenile ossifying fibroma warrant their classification as 2 distinct clinicopathologic entities. PMID- 11925540 TI - Castleman's disease of the buccal mucosa: report of a case and review of the literature of head and neck cases. AB - A case of Castleman's disease occurring in the buccal mucosa is described. An 84 year-old woman noticed that a mass in the left buccal mucosa that had been present for half a year. Computed tomography revealed a well-demarcated submucosal tumor, measuring 4.0 x 3.0 x 2.0 cm. The patient received no treatment at this time, and continued growth of the mass was observed. After incisional biopsy, the lesion was surgically removed. Histologically, the tumor consisted of an enlarged lymph node with conspicuous lymph follicles, in which vascular channels and deposits of eosinophilic material were noted. Laboratory examination showed an increase of serum antibody level of cytomegalovirus but of no other viruses. The patient was followed up for 1(1/2) years, with no clinical evidence of recurrence. This is the first report of Castleman's disease presenting in an oral site. PMID- 11925541 TI - Cementoblastoma: an innocuous neoplasm? A clinicopathologic study of 44 cases and review of the literature with special emphasis on recurrence. AB - The clinicopathologic features of 44 cases of cementoblastoma were analyzed and compared with those of 74 cases from the literature with special emphasis on the clinical behavior, treatment, and recurrence rate of these relatively rare benign odontogenic neoplasms. The cases in the current series were accessioned at the Armed Forces Institute of Pathology, Washington, DC. Patient ages at diagnosis ranged from 8 years to 44 years, with a mean age of 20.7 years. The tumors affected 30 males and 14 females. The mandible was the site of occurrence in 31 cases (70.4%), with the mandibular first molar the most common tooth involved. Radiographically, more than 90% of the tumors presented as well-defined radiopaque or mixed-density masses confluent with the tooth root(s) and surrounded by a radiolucent rim. Two lesions were radiolucent. Microscopically, cementoblastomas share similar features with osteoblastomas but are unique because of their physical attachment to the tooth root(s). Follow-up was obtained in 35 of 44 cases, with a mean follow-up interval of 5.5 years. Recurrence was documented in 13 cases (37.1%), in contrast to the literature, where only 2 of 34 cases (5.9%) with adequate follow-up recurred. Jaw expansion and perforation of the cortex were noted in a higher percentage of recurrent than nonrecurrent tumors. Because recurrence and continued growth are possible if lesional tissue remains after initial surgery, appropriate treatment should consist of removal of the lesion along with the affected tooth or teeth, followed by thorough curettage or peripheral ostectomy. PMID- 11925542 TI - A comparison of phosphor-plate digital images with conventional radiographs for the perceived clarity of fine endodontic files and periapical lesions. AB - OBJECTIVE: We sought to compare digital images with radiographs for the perceived clarity of small endodontic file tips at 2 different working lengths, as well as for the visualization of periapical bone lesions. STUDY DESIGN: Standardized conventional radiographic and phosphor-plate digital images were taken of 20 extracted permanent mandibular molars with 06 K-files placed in the distal root canal either 2 mm short or flush with the apical foramen. Similar images were obtained from mandibles with teeth that demonstrated large (n = 10) or small (n = 10) periapical lesions. Four evaluators ranked the clarity of the digital image with that of the radiograph. Results were analyzed by using the 2-sided sign test, ordinal logistic regression, and the kappa test. RESULTS: The perceived clarity of an endodontic file tip, at any position, and of a small or large periapical lesion was significantly (P < .01) less on all digital images compared with conventional films. CONCLUSION: Evaluator ratings indicated that the perceived clarity of fine endodontic files and periapical lesions was significantly less with phosphor-plate digital images than with conventional radiographs. PMID- 11925543 TI - Dental CT evaluation of mandibular first premolar root configurations and canal variations. AB - OBJECTIVES: Routine dental CT scans were used to describe mandibular first premolar root configurations and canal variations. STUDY DESIGN: One hundred twenty dental CT examinations were evaluated for mandibular first premolar root configurations and canal variations regarding shape of root and root canal, incidence of multiple canals, and level of bifurcation. RESULTS: A total of 17 teeth in 12 patients showed mesial invagination of the root of the mandibular first premolar. One root displayed 3 canals with 3 apical foramina. In 2 teeth, a single canal divided into 2 canals, but merged into 1 apical foramen. One root showing 2 root canals finally divided into 2 roots near the apex. Thirteen roots had 2 canals and 2 apical foramina. The distance from the cementoenamel junction to the level of bifurcation was between 4 and 13 mm (mean, 7.4 mm). CONCLUSION: The occurrence of incidentally found mandibular first premolar root variations should be an important component of dental CT reports. PMID- 11925544 TI - Comparison of different gutta-percha root filling techniques: Thermafil, Quick fill, System B, and lateral condensation. AB - OBJECTIVES: In this in vitro study, the core (gutta-percha or gutta-percha and carrier)/sealer ratio and the sealing ability were calculated for 4 different gutta-percha techniques: Thermafil, JS Quick-Fill, System B, and lateral condensation. STUDY DESIGN: One hundred single-rooted teeth were instrumented and obturated. Forty of the teeth were embedded in resin and sectioned horizontally at 1, 2,3, and 4 mm from the anatomic apex. Photographs were taken of each section, and the total area of the canal filled with core material or sealer was calculated. Sixty of the teeth were kept in wet storage for 90 days, after which the teeth were immersed in 2% methylene blue for 48 hours. The length of dye penetration was measured. RESULTS: Roots obturated with Thermafil and JS Quick Fill contained significantly more core material than did those obturated with System B or with lateral condensation. The lateral condensation technique had the lowest core/sealer ratio (P < .05). Roots using Thermafil and Quick-Fill showed significantly less dye leakage than did the lateral condensation technique. CONCLUSIONS: Through use of these techniques in vitro, Thermafil and JS Quick Fill with carrier and System B were found to be superior to the lateral condensation technique in terms of core/sealer ratio. Thermafil and Quick-Fill were superior to lateral condensation in terms of dye leakage. PMID- 11925545 TI - A comparison of D-, E-, and F-speed conventional intraoral radiographic films in endodontic measurement. AB - OBJECTIVES: We sought to compare observer measurement error and subjective ratings for International Standards Organization D-, E-, and F-speed direct exposure dental x-ray films used to determine endodontic working lengths. METHODS: Radiographs were exposed to achieve a standard background density of approximately 2.0. Three human maxillary posterior teeth for which No. 10 K-files had been placed at various lengths within 4 canals were studied in a cadaver section. A total of 30 radiographs were used (10 from each film type). Five licensed dentists excluding all authors viewed the radiographs independently and in random sequence. Distances from the file tips to the radiographic apices were measured and were compared with known lengths for error determination. The observing dentists also subjectively rated the images as desirable or undesirable. Statistical methods included a 3-factor ANOVA with Tukey honestly significant difference post hoc analysis to compare objective measurements and chi-square with respect to subjective ratings. RESULTS: No significant differences were attributable to film speed grouping or observers (P > .05); however, there was a specimen effect in that a significant difference was found in measurement accuracy among the 4 canals (P < .05). Differences in subjective ratings for the 3 film types were not statistically significant (P > .05). CONCLUSION: All 3 film types were similar in objective and subjective ratings. This being the case, the faster film is preferred to minimize the radiation dose to the patient. PMID- 11925546 TI - Evaluation of tuned-aperture computed tomography in the detection of simulated periodontal defects. AB - OBJECTIVE: The objective of this study was to compare the diagnostic efficacy of tuned-aperture computed tomography (TACT) and conventional film for the detection of simulated periodontal defects. STUDY DESIGN: Periodontal defects were created in 15 premolar-molar interproximal sites selected from 8 cadaver jaw segments. Ten observers viewed 45 image pairs (baseline and follow up) in 4 TACT modalities and film to assess the presence of defect. The TACT modalities were 1st and 2nd generation prototype systems (TACT-1 and TACT-2) with circular (C) and random (R) distribution patterns of source projection (TACT-1C, TACT-1R, TACT-2C, TACT-2R). Observer performance and related factors were analyzed by using receiver operating characteristic and analysis of variance. RESULTS: Mean A(z) values were 0.64 for film, 0.74 for TACT-1C, 0.82 for TACT-1R, 0.64 for TACT-2C, and 0.69 for TACT-2R. TACT-1R was significantly better than film (P < .001), TACT-2C (P < .001) and TACT-2R (P = .007). CONCLUSION: TACT-1R provided the best diagnostic performance in the detection of simulated periodontal defects among the 5 modalities compared in this study. PMID- 11925547 TI - Change of angiostructure and hemodynamics in lymph node metastases in rabbits. AB - OBJECTIVES: Experimental models of lymph node metastasis were developed for Doppler sonography. METHODS: Cervical lymph node (LN) metastasis was induced by the implantation of a VX-2 tumor on the oral floor of 10 rabbits. Twenty metastatic LNs were observed weekly by power Doppler sonography, and the presence of an avascular area and the peripheral vessels were evaluated. The time-averaged maximum velocity (TAMx) at the hilum was also measured. RESULTS: The percentage of the metastatic LNs presenting with an avascular area increased over time. The peripheral vessels were detected in 14 of 20 LNs. The peak time of the TAMx significantly correlated to that of the initial detection of the avascular area. CONCLUSIONS: In the beginning, metastatic LNs were depicted as hypervascular structures while the TAMx at the hilum increased. Later, an avascular area was detected and the TAMx at the hilum decreased, which resulted in a blood supply to the node from the peripheral vessels. PMID- 11925548 TI - Inferior alveolar nerve injury related to mandibular third molar surgery: an unusual case presentation. AB - Perforation of the lower third molar roots by the inferior alveolar nerve is uncommon and can be difficult to determine by conventional radiographic methods. Presented is a case of perforation that was treated by coronectomy, and showed an unusual complication in that the retained root erupted, moving the canal with it. The radiographic assessment of root perforation and the imaging modalities used to assess such cases are discussed. PMID- 11925549 TI - Diagnosis and radiologic manifestations of malignant dumbbell tumors of the parotid gland: review and 2 case reports. AB - Primary malignant dumbbell tumors represent a special growth type of deep-lobe parotid neoplasm. Because they are located in an anatomically limited space between the skull base and the mandibular ramus, they do not cause any but a few nonspecific symptoms and may metastasize into the neck or distant organs before the primary tumor has ever been diagnosed. This article describes 2 cases that first presented as asymptomatic carcinoma unknown primary syndrome and were later discovered to be malignant dumbbell tumors of the parotid gland. Because adequate evaluation is essential for the subsequent management, several diagnostic methods available for the differential diagnosis of this tumor are discussed; we compare these according to their relevance to the therapeutic procedure. Magnetic resonance imaging was found to be the most appropriate imaging technique because of its ability to help us better differentiate between tumor and surrounding tissue. Computed tomography and single photon emission computed tomography can help us identify involvement of the skull base and mandibular ramus. PMID- 11925550 TI - Error patterns and observer variations in the high-resolution ultrasonography imaging evaluation of the disk position of the temporomandibular joint. AB - OBJECTIVES: The purpose of this study was (1) to describe errors in the interpretation of high-resolution ultrasonography (HR-US) images of the temporomandibular joint (TMJ) and analyze the discrepancies between HR-US imaging and diagnostic magnetic resonance imaging findings, and (2) to measure interobserver and intraobserver variations with respect to reporting HR-US images of the TMJ. STUDY DESIGN: The sonographic reports of 208 patients aged 13 to 78 years who had a prospective clinical diagnosis of TMJ internal derangement were correlated with magnetic resonance imaging results. Prospective and retrospective readings were used to classify types of disk displacement. Assessment of observer variation was based on a set of 200 HR-US images in 50 randomly selected patients. The images were interpreted independently by 2 trained radiologists at 2 sessions, 1 to 2 weeks apart. RESULTS: Prospective interpretation of TMJ HR-US images at the closed-mouth position had a sensitivity of 80%, specificity of 87%, and accuracy of 82%. For images at the maximum mouth-opening position, the sensitivity was 68%, specificity 93%, and accuracy 82%. The number of diagnostic errors at retrospective analysis was 60.3% (n = 73) of the number at prospective analysis. The intraobserver agreement for reporting the disk position at the closed-mouth position was 93% and at the maximum mouth-opening position was 88%. The agreement between the 2 observers was 89% on the position of the disk at the closed-mouth position and 84% at the maximum mouth-opening position. CONCLUSIONS: The high diagnostic accuracy, together with the low observer variation reported in this study, suggest that HR-US of the TMJ is a valuable imaging technique for assessment of disk displacement. Many errors that occur at prospective analysis didn't occur at retrospective analysis. PMID- 11925551 TI - Wag the dogma. PMID- 11925552 TI - Ten years on. PMID- 11925553 TI - A roundabout route to gene therapy. PMID- 11925554 TI - Putting tubby on the MAP. PMID- 11925555 TI - Bringing order to organogenesis. PMID- 11925556 TI - Evaluating adaptive evolution. PMID- 11925557 TI - Mitochondria, from cell death to proliferation. PMID- 11925560 TI - Biotechnology patents: strategies for meeting economic and ethical concerns. PMID- 11925561 TI - Inter-mitochondrial complementation of mtDNA mutations and nuclear context. PMID- 11925563 TI - A bivalent Huntingtin binding peptide suppresses polyglutamine aggregation and pathogenesis in Drosophila. AB - Huntington disease is caused by the expansion of a polyglutamine repeat in the Huntingtin protein (Htt) that leads to degeneration of neurons in the central nervous system and the appearance of visible aggregates within neurons. We have developed and tested suppressor polypeptides that bind mutant Htt and interfere with the process of aggregation in cell culture. In a Drosophila model, the most potent suppressor inhibits both adult lethality and photoreceptor neuron degeneration. The appearance of aggregates in photoreceptor neurons correlates strongly with the occurrence of pathology, and expression of suppressor polypeptides delays and limits the appearance of aggregates and protects photoreceptor neurons. These results suggest that targeting the protein interactions leading to aggregate formation may be beneficial for the design and development of therapeutic agents for Huntington disease. PMID- 11925564 TI - Disruption of an SF2/ASF-dependent exonic splicing enhancer in SMN2 causes spinal muscular atrophy in the absence of SMN1. AB - Alteration of correct splicing patterns by disruption of an exonic splicing enhancer may be a frequent mechanism by which point mutations cause genetic diseases. Spinal muscular atrophy results from the lack of functional survival of motor neuron 1 gene (SMN1), even though all affected individuals carry a nearly identical, normal SMN2 gene. SMN2 is only partially active because a translationally silent, single-nucleotide difference in exon 7 causes exon skipping. Using ESE motif-prediction tools, mutational analysis and in vivo and in vitro splicing assays, we show that this single-nucleotide change occurs within a heptamer motif of an exonic splicing enhancer, which in SMN1 is recognized directly by SF2/ASF. The abrogation of the SF2/ASF-dependent ESE is the basis for inefficient inclusion of exon 7 in SMN2, resulting in the spinal muscular atrophy phenotype. PMID- 11925565 TI - Rescue of a deficiency in ATP synthesis by transfer of MTATP6, a mitochondrial DNA-encoded gene, to the nucleus. AB - A T-->G transversion at nt 8993 in mitochondrial DNA of MTATP6 (encoding ATPase 6 of complex V of the respiratory chain) causes impaired mitochondrial ATP synthesis in two related mitochondrial disorders: neuropathy, ataxia and retinitis pigmentosa and maternally inherited Leigh syndrome. To overcome the biochemical defect, we expressed wildtype ATPase 6 protein allotopically from nucleus-transfected constructs encoding an amino-terminal mitochondrial targeting signal appended to a recoded ATPase 6 gene (made compatible with the universal genetic code) that also contained a carboxy-terminal FLAG epitope tag. After transfection of human cells, the precursor polypeptide was expressed, imported into and processed within mitochondria, and incorporated into complex V. Allotopic expression of stably transfected constructs in cytoplasmic hybrids (cybrids) homoplasmic with respect to the 8993T-->G mutation showed a significantly improved recovery after growth in selective medium as well as a significant increase in ATP synthesis. This is the first successful demonstration of allotopic expression of an mtDNA-encoded polypeptide in mammalian cells and could form the basis of a genetic approach to treat a number of human mitochondrial disorders. PMID- 11925567 TI - Adaptive evolution of a duplicated pancreatic ribonuclease gene in a leaf-eating monkey. AB - Although the complete genome sequences of over 50 representative species have revealed the many duplicated genes in all three domains of life, the roles of gene duplication in organismal adaptation and biodiversity are poorly understood. In addition, the evolutionary forces behind the functional divergence of duplicated genes are often unknown, leading to disagreement on the relative importance of positive Darwinian selection versus relaxation of functional constraints in this process. The methodology of earlier studies relied largely on DNA sequence analysis but lacked functional assays of duplicated genes, frequently generating contentious results. Here we use both computational and experimental approaches to address these questions in a study of the pancreatic ribonuclease gene (RNASE1) and its duplicate gene (RNASE1B) in a leaf-eating colobine monkey, douc langur. We show that RNASE1B has evolved rapidly under positive selection for enhanced ribonucleolytic activity in an altered microenvironment, a response to increased demands for the enzyme for digesting bacterial RNA. At the same time, the ability to degrade double-stranded RNA, a non-digestive activity characteristic of primate RNASE1, has been lost in RNASE1B, indicating functional specialization and relaxation of purifying selection. Our findings demonstrate the contribution of gene duplication to organismal adaptation and show the power of combining sequence analysis and functional assays in delineating the molecular basis of adaptive evolution. PMID- 11925566 TI - Microtubule-associated protein 1A is a modifier of tubby hearing (moth1). AB - Once a mutation in the gene tub was identified as the cause of obesity, retinal degeneration and hearing loss in tubby mice, it became increasingly evident that the members of the tub gene family (tulps) influence maintenance and function of the neuronal cell lineage. Suggested molecular functions of tubby-like proteins include roles in vesicular trafficking, mediation of insulin signaling and gene transcription. The mechanisms through which tub functions in neurons, however, have yet to be elucidated. Here we report the positional cloning of an auditory quantitative trait locus (QTL), the modifier of tubby hearing 1 gene (moth1), whose wildtype alleles from strains AKR/J, CAST/Ei and 129P2/OlaHsd protect tubby mice from hearing loss. Through a transgenic rescue experiment, we verified that sequence polymorphisms in the neuron-specific microtubule-associated protein 1a gene (Mtap1a) observed in the susceptible strain C57BL/6J (B6) are crucial for the hearing-loss phenotype. We also show that these polymorphisms change the binding efficiency of MTAP1A to postsynaptic density molecule 95 (PSD95), a core component in the cytoarchitecture of synapses. This indicates that at least some of the observed polymorphisms are functionally important and that the hearing loss in C57BL/6J-tub/tub (B6-tub/tub) mice may be caused by impaired protein interactions involving MTAP1A. We therefore propose that tub may be associated with synaptic function in neuronal cells. PMID- 11925568 TI - c-fos regulates neuronal excitability and survival. AB - Excitotoxicity is a process in which glutamate or other excitatory amino acids induce neuronal cell death. Accumulating evidence suggests that excitotoxicity may contribute to human neuronal cell loss caused by acute insults and chronic degeneration in the central nervous system. The immediate early gene (IEG) c-fos encodes a transcription factor. The c-Fos proteins form heterodimers with Jun family proteins, and the resulting AP-1 complexes regulate transcription by binding to the AP-1 sequence found in many cellular genes. Emerging evidence suggests that c-fos is essential in regulating neuronal cell survival versus death. Although c-fos is induced by neuronal activity, including kainic acid induced seizures, whether and how c-fos is involved in excitotoxicity is still unknown. To address this issue, we generated a mouse in which c-fos expression is largely eliminated in the hippocampus. We found that these mutant mice have more severe kainic acid-induced seizures, increased neuronal excitability and neuronal cell death, compared with control mice. Moreover, c-Fos regulates the expression of the kainic acid receptor GluR6 and brain-derived neurotrophic factor (BDNF), both in vivo and in vitro. Our results suggest that c-fos is a genetic regulator for cellular mechanisms mediating neuronal excitability and survival. PMID- 11925569 TI - Tenascin-X deficiency mimics Ehlers-Danlos syndrome in mice through alteration of collagen deposition. AB - Tenascin-X is a large extracellular matrix protein of unknown function. Tenascin X deficiency in humans is associated with Ehlers-Danlos syndrome, a generalized connective tissue disorder resulting from altered metabolism of the fibrillar collagens. Because TNXB is the first Ehlers-Danlos syndrome gene that does not encode a fibrillar collagen or collagen-modifying enzyme, we suggested that tenascin-X might regulate collagen synthesis or deposition. To test this hypothesis, we inactivated Tnxb in mice. Tnxb-/- mice showed progressive skin hyperextensibility, similar to individuals with Ehlers-Danlos syndrome. Biomechanical testing confirmed increased deformability and reduced tensile strength of their skin. The skin of Tnxb-/- mice was histologically normal, but its collagen content was significantly reduced. At the ultrastructural level, collagen fibrils of Tnxb-/- mice were of normal size and shape, but the density of fibrils in their skin was reduced, commensurate with the reduction in collagen content. Studies of cultured dermal fibroblasts showed that although synthesis of collagen I by Tnxb-/- and wildtype cells was similar, Tnxb-/- fibroblasts failed to deposit collagen I into cell-associated matrix. This study confirms a causative role for TNXB in human Ehlers-Danlos syndrome and suggests that tenascin-X is an essential regulator of collagen deposition by dermal fibroblasts. PMID- 11925571 TI - The relief of suffering in mental illness. PMID- 11925572 TI - The nature of psychiatric nursing: the intersection of paradigm, evolution, and history. AB - Psychiatric nursing, while vibrant and applicable once, is predicated on an approach to care and an understanding of mental illness that reflects an older time. Psychiatric nursing is in the precarious position of having an aging paradigm of practice, rooted in older knowledge and beliefs and recalcitrant to new knowledge and new realities of clinical practice. This article highlights paradigmatic assumptions and identifies the two dominant knowledge structures that struggle to coexist within that paradigm. Practical consequences of practicing psychiatric nursing with an aging paradigm are reviewed and a beginning point for the development of a new paradigm is presented. PMID- 11925570 TI - Genomic disorders on 22q11. AB - The 22q11 region is involved in chromosomal rearrangements that lead to altered gene dosage, resulting in genomic disorders that are characterized by mental retardation and/or congenital malformations. Three such disorders-cat-eye syndrome (CES), der(22) syndrome, and velocardiofacial syndrome/DiGeorge syndrome (VCFS/DGS)-are associated with four, three, and one dose, respectively, of parts of 22q11. The critical region for CES lies centromeric to the deletion region of VCFS/DGS, although, in some cases, the extra material in CES extends across the VCFS/DGS region. The der(22) syndrome region overlaps both the CES region and the VCFS/DGS region. Molecular approaches have revealed a set of common chromosome breakpoints that are shared between the three disorders, implicating specific mechanisms that cause these rearrangements. Most VCFS/DGS and CES rearrangements are likely to occur by homologous recombination events between blocks of low-copy repeats (e.g., LCR22), whereas nonhomologous recombination mechanisms lead to the constitutional t(11;22) translocation. Meiotic nondisjunction events in carriers of the t(11;22) translocation can then lead to offspring with der(22) syndrome. The molecular basis of the clinical phenotype of these genomic disorders has also begun to be addressed. Analysis of both the genomic sequence for the 22q11 interval and the orthologous regions in the mouse has identified >24 genes that are shared between VCFS/DGS and der(22) syndrome and has identified 14 putative genes that are shared between CES and der(22) syndrome. The ability to manipulate the mouse genome aids in the identification of candidate genes in these three syndromes. Research on genomic disorders on 22q11 will continue to expand our knowledge of the mechanisms of chromosomal rearrangements and the molecular basis of their phenotypic consequences. PMID- 11925573 TI - The PLUS intervention: a pilot test with caregivers of depressed older adults. AB - The PLUS Nursing Intervention, which is aimed at caregivers of elderly persons with depression and designed to increase caregiver personal resources, respond to caregiver learning/skill development needs, address caregiver unanticipated needs, and assist with caregiver stress/illness management, was pilot tested for efficacy. Thirteen families were assigned to the PLUS group and 12 families to the standard home care control group. There were no significant outcome differences between the two groups. However, caregivers who received the PLUS intervention made significantly more improvements over Standard Home Care caregivers when patients made functional improvements. Findings suggest that patient functioning might be a better predictor of long-term caregiver outcomes than psychiatric symptoms. PMID- 11925574 TI - Current concepts in body dysmorphic disorder. AB - Body dysmorphic disorder (BDD) is a potentially debilitating psychiatric illness characterized by intense preoccupation with an imagined or slight defect in physical appearance. Until recently, it has gone largely unrecognized and did not appear in the Diagnostic and Statistical Manual of Mental Disorders until 1987. Improved methods of assessment and treatment of BDD have increased interest in this disorder. This article reviews current literature on BDD, including the similarities between BDD and obsessive-compulsive disorder (OCD), current assessment and treatment strategies, and implications for clinical practice and future research. PMID- 11925575 TI - Temporal regularities in physical control at a state psychiatric hospital. AB - Physical control (mechanical restraint and locked seclusion) of psychiatric patients has received increasing scrutiny, and better understanding of the timing of physical control would allow for more informed choices about its implementation. This study examined temporal regularities in the use of physical control at a general psychiatric hospital over a period of 8.6 years. As hypothesized, physical control was not randomly distributed through time but instead showed positive autocorrelation; yearly, weekly, and daily cycles; and higher frequency at patients' smoking, meal, and medication times. These results are discussed in the context of physical control research, theory, and policy. PMID- 11925576 TI - Methodological challenges encountered in conducting a comparative study of psychiatric nursing education approaches in two African countries: Botswana and Nigeria. AB - This article discusses some methodological challenges encountered when conducting a comparative study of psychiatric nursing education approaches adopted in two sub-Saharan African countries - Botswana and Nigeria. The article identifies the methodological problems encountered and ways in which these challenges were addressed, including the triangulation of data collection strategies guided by Lewin's force field analysis and utilizing curriculum evaluation checklists. Data collection sources included a self-reporting questionnaire completed by psychiatric nurse educators, focus group discussions with practicing psychiatric nurses, interviews with representatives of the nursing regulatory bodies, and analyses of documents and records related to psychiatric nursing education in the two selected countries. Recommendations are provided for conducting further comparative studies that could enhance the globalization of psychiatric nursing education within the African continent, and possibly across other countries' borders as well. PMID- 11925577 TI - Are you comfortable enough to reach out? PMID- 11925578 TI - The nurse as patient advocate. PMID- 11925579 TI - Accuracy of references in three critical care nursing journals. AB - Reference lists serve as a resource for the reader and provide support for the author's ideas. Errors in references reflect poorly on the author and journal and may prevent locating the referenced articles. The purpose of this study was to determine the number and types of errors in references in 3 critical care nursing journals: Journal of PeriAnesthesia Nursing, American Journal of Critical Care, and Critical Care Nurse. There were 2,444 citations in 13 issues of these journals during the study period. Ten percent of the references from each journal was selected randomly and analyzed for accuracy. Of the 244 references examined, 56 contained errors for an overall error rate of 22.9%. Major errors, such as misspelled or omitted author names and initials, occurred in 19.6% of the references. Minor errors, including non-first page discrepancies, occurred in 4.5% of the references. Errors in author names combined with incorrect or missing volume or issue numbers were the 2 most common errors, accounting for 61% of errors. The overall error rate for these 3 critical care nursing journals was lower than the rates reported for other nursing journals. PMID- 11925580 TI - Statistical power analysis to estimate how many months of data are required to identify PACU staffing to minimize delays in admission from ORs. AB - When each nurse in the Phase I setting is caring for the maximum number of patients allowed by hospital staffing standards (typically 2 per ASPAN standards), patients may have to be held in the OR until a PACU nurse becomes available. Previously, the authors described a statistical method to determine the process of scheduling existing nurses without increasing staffing hours (Dexter et al. Anesth Analg. 92:947-949, 2001). The end result was to minimize the percentage of future workdays during which at least one patient would wait in his or her OR for Phase I PACU admission. In this study, the authors performed a statistical power analysis to determine how many months of PACU workload data are needed to optimize PACU staffing by using this "set covering" algorithm. One year (232 workdays) of data was available from a PACU employing up to 10 nurses working a total of 72 clinical hours a day. The data were divided into 2 subsets. Using the first subset, which varied in size between 20 and 140 days of data, the authors identified the optimal staffing solutions. These solutions were tested on the second subset of data. This process then was repeated thousands of times. There was a marked improvement in the performance of the staffing solutions at preventing "PACU hold" by increasing from 20 to 80 historical workdays of data, a slight but statistically significant improvement between 80 and 100 workdays, but no significant improvement in further increasing the number of workdays of data. PACU nurse managers should use at least 4 months of data when choosing a staffing solution to minimize the chance of patients waiting in ORs for PACU admission. Tampering with PACU staffing more often than every 4 months is unlikely to result in improvements in OR efficiency and may harm recruitment and retention of nursing staff. PMID- 11925581 TI - Sensorineural hearing loss after select procedures. AB - Hearing is one of the most significant senses; therefore, any loss can be frightening. Previous studies have found that certain surgical and medical procedures can result in hearing loss. Cardiac surgical procedures are most commonly associated with hearing loss owing to the use of the cardiopulmonary bypass pump. Other noncardiac, nonotologic surgical procedures also may result in hearing loss. Additionally, medical procedures such as myelogram and lumbar puncture have been associated with hearing loss. Studies suggest that hearing loss can result any time cerebrospinal fluid is leaked. This report highlights the effect of dural puncture on hearing loss. The implication that any dural puncture procedure may result in a cerebrospinal fluid leak and subsequent hearing loss is a significant clinical topic. Perianesthesia nurses are advised to educate themselves and their patients regarding this complication. PMID- 11925582 TI - Comfort care: a framework for perianesthesia nursing. AB - The Theory of Comfort and its application to the specialty of perianesthesia nursing are presented in this article. Comfort Theory is relevant to this specialty because patient comfort is cited as a goal in its standards of care and is an established value for many nurses in their perianesthesia practice. The technical definition of comfort, developed from reviews of multidisciplinary literature on comfort and nursing literature on holism, is "the state of being strengthened by having needs for relief, ease, and transcendence met in 4 contexts of experience (physical, psychospiritual, sociocultural, and environmental)." (Kolcaba K. Adv Nurs Sci. 15:1-10, 1992). Because comfort has many interrelated aspects, interventions designed to enhance comfort in one aspect can have a larger effect on total comfort than expected. Particular emphasis is placed on interventions to treat anxiety, because this discomfort can be severe and can negatively affect physiologic function. Goals and comfort measures for each phase of perianesthesia care are presented, followed by application of Comfort Theory for research. PMID- 11925583 TI - Taking a look at the Joint Commission. PMID- 11925584 TI - Mentoring for perianesthesia nurses. PMID- 11925585 TI - Safety concerns with meperidine. PMID- 11925586 TI - The three faces of anthrax. PMID- 11925587 TI - Notes from the 2001 American Society of Anesthesiologists annual meeting. PMID- 11925588 TI - Acute coronary syndromes. PMID- 11925589 TI - We are women, hear us roar! PMID- 11925591 TI - The abnormal expression of retinoic acid receptor-beta, p 53 and Ki67 protein in normal, premalignant and malignant esophageal tissues. AB - AIM: Esophageal cancer remains a significant health problem worldwide. It is important to investigate alterations in expression of retinoic acid receptor beta, p 53 and Ki67 proteins in esophageal carcinogenesis. METHODS: To find biomarkers for early identification of esophageal cancer, we analyzed the retinoic acid receptor-beta, p 53 protein and the proliferation marker Ki67 in surgical specimens of normal, mildly, and severely dysplastic and malignant esophageal tissues by in situ hybridization of RNA and immunohistochemistry. RESULTS: RAR-beta was expressed in 94.3%(33/35) of normal mucosae, 67.8%(19/28) of the mild, 58.1% (18/31) of the severe lesions and 53.2%(116/218) of tumor samples. RAR-beta mRNA was expressed in 62.7%(42/67), 55.1%(43/78) and 29.2%(7/24) of well, moderated and poorly differentiated SSCs. The p 53 and Ki67 proteins were 5.9%(2/34) of the normal mucosa. P53 and Ki67 stained positively in 10.7%(3/28) and 21.4% (6/28) of mild dysplasia, and 51.6%(16/31) and 58.1% (18/31) of severely dysplasia respectively. Samples from esophageal cancer showed no higher levers of p 53 and Ki67 expression than seen in severely dysplastic lesions. There was significant difference of RAR-beta p 53 and Ki67 expression between normal mucosa and dysplastic tissue or esophageal cancer. CONCLUSION: Loss of RAR-beta expression and accumulation of p 53 and Ki67 proteins may serve as biomarkers for early identification of esophageal cancer in the high-risk populations. PMID- 11925590 TI - The prognostic significance of clinical and pathological features in hepatocellular carcinoma. AB - The prognosis of patients with HCC still remains dismal. The life expectancy of HCC patients is hard to predict because of the high possibility of postoperative recurrence. Many factors, such as patient's general conditions, macroscopic tumor morphology, as well as tumor histopathology features, have been proven of prognostic significance. Female HCC patient often has a better prognosis than male patient, which might be due to the receptor of sex hormones. Younger patients often have tumors with higher invasiveness and metastatic potentials, and their survival and prognosis are worse than the older ones. Co-existing hepatitis status and hepatic functional reserve have been confirmed as risk factors for recurrence. Serum alpha-fetoprotein (AFP) is useful not only for diagnosis, but also as a prognostic indicator for HCC patients. AFP mRNA has been proposed as a predictive marker of HCC cells disseminated into the circulation and for metastatic recurrence. Many pathologic features, such as tumor size, number, capsule state, cell differentiation, venous invasion, intrahepatic spreading, and advanced pTNM stage, are the best-established risk factors for recurrence and important aspects affecting the prognosis of patients with HCC. Marked inflammatory cell infiltration in the tumor could predict a better prognosis. Clinical stage is still the most important factor influencing on the prognosis. Extratumor spreading and lymph nodal metastasis are independent predictors for poor outcome. Some new predictive systems have recently been proposed. Different strategies of treatment might have significant different effects on the patients' prognosis. To date, surgical resection is still the only potentially curative treatment for HCC, including localized postoperative recurrences. Extent of resection, blood transfusion, occlusion of porta hepatis, and blood loss affect the survival and prognosis of HCC patients. Regional therapies provide alternative ways to improve the prognosis of HCC patients who have no opportunity to receive surgical treatment or postoperative recurrence. The combination of these treatment modalities is hopeful to further improve the prognosis. The efficacies of neoadjuvant (preoperative) or adjuvant (postoperative) chemotherapy or chemoembolization in preventing recurrence and on the HCC prognosis still remain great controversy, and deserve further evaluation. Biotherapy, including IFN-alpha therapy, will play more important role in preventing recurrence and metastasis of HCC after operation. PMID- 11925592 TI - Transcription factor EGR-1 inhibits growth of hepatocellular carcinoma and esophageal carcinoma cell lines. AB - AIM: The transcription factor EGR-1 (early growth response gene-1) plays an important role in cell growth, differentiation and development. It has identified that EGR-1 has significant transformation suppression activity in some neoplasms, such as fibrosarcoma, breast carcinoma. This experiment was designed to investigate the role of egr-1 in the cancerous process of hepatocellular carcinoma (HCC) and esophageal carcinoma (EC), and then to appraise the effects of EGR-1 on the growth of these tumor cells. METHODS: Firstly, the transcription and expression of egr-1 in HCC and EC, paracancerous tissues and their normal counterpart parts were detected by in situ hybridization and immunohistochemistry, with normal human breast and mouse brain tissues as positive controls. Egr-1 gene was then transfected into HCC (HHCC, SMMC7721) and EC (ECa109) cell lines in which no egr-1 transcription and expression were present. The cell growth speed, FCM cell cycle, plate clone formation and tumorigenicity in nude mice were observed and the controls were the cell lines transfected with vector only. RESULTS: Little or no egr-1 transcription and expression were detected in HCC, EC and normal liver tissues. The expression of egr-1 were found higher in hepatocellular paracancerous tissue (transcription level P=0.000; expression level P=0.143, probably because fewer in number of cases) and dysplastic tissue of esophageal cancer (transcription level P=0.000; expression level P=0.001). The growth rate of egr-1-transfected HHCC (HCC cell line) cells and ECa109 (EC cell line) cells was much slower than that of the controls. The proportion of S phase cell, clone formation and tumorigenicity were significantly lower than these of the controls' (decreased 45.5% in HHCC cells and 34.1% in ECa109 cells; 46.6% and 41.8%; 80.4% and 72.6% respectively). There were no obvious differences between SMMC7721 (HCC) egr-1-transfected cells and the controls with regard to the above items. CONCLUSION: The decreased expression of egr-1 might play a role in the dysregulation of normal growth in the cancerous process of HCC and EC. Egr-1 gene of transfected HHCC and ECa109 cells showed obvious suppression of the cell growth and malignant phenotypes, but no suppression in SMMC7721 (HCC cell line) cells. PMID- 11925593 TI - Construction of cDNA representational difference analysis based on two cDNA libraries and identification of garlic inducible expression genes in human gastric cancer cells. AB - AIM: To elucidate molecular mechanism of chemopreventive efficacies of garlic against human gastric cancer (HGC). METHODS: HGC cell line BGC823 was treated with Allitridi (a kind of garlic extract) and Allitridi-treated and parental BGC823 cDNA libraries were constructed respectively by using lambdaZAP II vector. cDNA Representational Difference Analysis (cDNA RDA) was performed using Bam H I cutting-site and abundant cDNA messages provided by the libraries. Northern blot analysis was applied to identify the obtained difference products. RESULTS: Two specific cDNA fragments were obtained and characterized to be derived from homo sapiens folate receptor alpha (FRalpha) gene and calcyclin gene respectively. Northern blot results showed a 4-fold increase in FRalpha gene expression level and 9-fold increase in calcyclin mRNA level in BGC823 cells after Allitridi treatment for 72h. CONCLUSION: The method of cDNA RDA based on cDNA libraries combines the high specificity of cDNA RDA with abundant cDNA messages in cDNA library; this expands the application of cDNA library and increases the specificity of cDNA RDA. Up-regulation of FRalpha gene and calcyclin gene expressions induced by Allitridi provide valuable molecular evidence for the efficacy garlic in treating HGC as well as other diseases. PMID- 11925594 TI - Applying a highly specific and reproducible cDNA RDA method to clone garlic up regulated genes in human gastric cancer cells. AB - AIM: To develop and optimize cDNA representational difference analysis (cDNA RDA) method and to identify and clone garlic up-regulated genes in human gastric cancer (HGC) cells. METHODS: We performed cDNA RDA method by using abundant double-stranded cDNA messages provided by two self-constructed cDNA libraries (Allitridi-treated and paternal HGC cell line BGC823 cells cDNA libraries respectively). Bam H I and Xho I restriction sites harbored in the library vector were used to select representations. Northern and Slot blots analyses were employed to identify the obtained difference products. RESULTS: Fragments released from the cDNA library vector after restriction endonuclease digestion acted as good marker indicating the appropriate digestion degree for library DNA. Two novel expressed sequence tags (ESTs) and a recombinant gene were obtained. Slot blots result showed a 8-fold increase of glia-derived nexin/protease nexin 1 (GDN/PN1) gene expression level and 4-fold increase of hepatitis B virus x interacting protein (XIP) mRNA level in BGC823 cells after Allitridi treatment for 72h. CONCLUSION: Elevated levels of GDN/PN1 and XIP mRNAs induced by Allitridi provide valuable molecular evidence for elucidating the garlic's efficacies against neurodegenerative and inflammatory diseases. Isolation of a recombinant gene and two novel ESTs further show cDNA RDA based on cDNA libraries to be a powerful method with high specificity and reproducibility in cloning differentially expressed genes. PMID- 11925595 TI - JTE-522-induced apoptosis in human gastric adenocarcinoma [correction of adenocarcinoma] cell line AGS cells by caspase activation accompanying cytochrome C release, membrane translocation of Bax and loss of mitochondrial membrane potential. AB - AIM: To investigate the role of the mitochondrial pathway in JTE-522-induced apoptosis and to investigate the relationship between cytochrome C release, caspase activity and loss of mitochondrial membrane potential (Deltapsim). METHODS: Cell culture, cell counting, ELISA assay, TUNEL, flow cytometry, Western blot and fluorometric assay were employed to investigate the effect of JTE-522 on cell proliferation and apoptosis in AGS cells and related molecular mechanism. RESULTS: JTE-522 inhibited the growth of AGS cells and induced the apoptosis. Caspases 8 and 9 were activated during apoptosis as judged by the appearance of cleavage products from procaspase and the caspase activities to cleave specific fluorogenic substrates. To elucidate whether the activation of caspases 8 and 9 was required for the apoptosis induction, we examined the effect of caspase specific inhibitors on apoptosis. The results showed that caspase inhibitors significantly inhibited the apoptosis induced by JTE-522. In addition, the membrane translocation of Bax and cytosolic release of cytochrome C accompanying with the decrease of the uptake of Rhodamin 123, were detected at an early stage of apoptosis. Furthermore, Bax translocation, cytochrome C release, and caspase 9 activation were blocked by Z-VAD.fmk and Z-IETD-CHO. CONCLUSION: The present data indicate a crucial association between activation of caspases 8, 9, cytochrome C release, membrane translocation of Bax, loss of Deltapsim and JTE-522-induced apoptosis in AGS cells. PMID- 11925596 TI - Effect of cis-9, trans-11-conjugated linoleic acid on cell cycle of gastric adenocarcinoma cell line (SGC-7901). AB - AIM: To determine the effect of cis -9, trans -11-conjugated linoleic acid (c9, t11-CLA) on the cell cycle of gastric cancer cells (SGC-7901) and its possible mechanism in inhibition cancer growth. METHODS: Using cell culture and immunocytochemical techniques, we examined the cell growth, DNA synthesis, expression of PCNA, cyclin A, B(1), D(1), p16(ink4a) and p21(cip/waf1) of SGC 7901 cells which were treated with various c9, t11-CLA concentrations (25, 50, 100 and 200 micromol.L(-1))of c 9, t 11-CLA for 24 and 48h, with a negative control (0.1% ethane). RESULTS: The cell growth and DNA synthesis of SGC-7901 cells were inhibited by c9, t11-CLA.SGC-7901 cells. Eight day after treatment with various concentrations of c9, t11-CLA mentioned above, the inhibition rates were 5.92%, 20.15%, 75.61% and 82.44%, respectively and inhibitory effect of c9, t11-CLA on DNA synthesis (except for 25 micromol.L, 24h) showed significantly less (3)H-TdR incorporation than that in the negative controls (P<0.05 and P<0.01). Immunocytochemical staining demonstrated that SGC-7901 cells preincubated in media supplemented with different c9, t11-CLA concentrations at various times significantly decreased the expressions of PCNA (the expression rates were 7.2-3.0%, 24h and 9.1-0.9% at 48h, respectively), Cyclin A (11.0-2.3%, 24h and 8.5-0.5%,48h), B(1) (4.8-1.8% at 24h and 5.5-0.6% at 48h)and D(1) (3.6 1.4% at 24h and 3.7%-0 at 48h) as compared with those in the negative controls(the expressions of PCNA, Cyclin A, B(1) and D(1) were 6.5% at 24h and 9.0% at 48h, 4.2% at 24h and 5.1% at 48h, 9.5% at 24h and 6.0% at 48h,respectively)(P<0.01), whereas the expressions of P16(ink4a) and P21(cip/waf1), cyclin-dependent kinases inhibitors(CDKI), were increased. CONCLUSION: The cell growth and proliferation of SGC-7901 cell is inhibited by c9, t11-CLA via blocking the cell cycle, with reduced expressions of cyclin A,B(1) and D(1) and enhanced expressions of CDKI(P16(ink4a) and p21(cip/waf1)). PMID- 11925597 TI - Methionine-dependence and combination chemotherapy on human gastric cancer cells in vitro. AB - AIM: To elucidate whether human primary gastric cancer and gastric mucosa epithelial cells in vitro can grow normally in a methionine (Met) depleted environment, i.e. Met-dependence, and whether Met-depleting status can enhance the killing effect of chemotherapy on gastric cancer cells. METHODS: Fresh human gastric cancer and mucosal tissues were managed to form monocellular suspensions, which were then cultured in the Met-free but homocysteine-containing (Met( )Hcy(+)) medium, with different chemotherapeutic drugs. The proliferation of the cells was examined by cell counter, flow cytometry (FCM) and microcytotoxicity assay (MTT). RESULTS: The growth of human primary gastric cancer cells in Met( )Hcy(+) was suppressed, manifested by the decrease of total cell counts [1.46 +/- 0.42 (x 10(9).L(-1)) in Met(-)Hcy(+) vs 1.64 +/-0.44(x 10(9).L(-1)) in Met(+)Hcy( ), P<0.01], the decline in the percentage of G(0)G(1) phase cells (0.69 +/- 0.24 in Met(-)Hcy(+) vs 0.80 +/- 0.18 in Met(+)Hcy(-), P<0.01) and the increase of S cells (0.24 +/- 0.20 in Met(-)Hcy(+) vs 0.17 +/- 0.16 in Met(+)Hcy(-), P<0.01); however, gastric mucosal cells grew normally. If Met(-)Hcy(+) medium was used in combination with chemotherapeutic drugs, the number of surviving gastric cancer cells dropped significantly. CONCLUSION: Human primary gastric cancer cells in vitro are Met-dependent; however, gastric mucosal cells have not shown the same characteristics. Met(-)Hcy(+) environment may strengthen the killing effect of chemotherapy on human primary gastric cancer cells. PMID- 11925598 TI - Effects of cryopreservation and phenylacetate on biological characters of adherent LAK cells from patients with hepatocellular carcinoma. AB - AIM: To improve the preparation of adherent lymphokine-activated killer (A-LAK) cells and to study the effects of cryopreservation and phenylacetate (PA) on biological characters of A-LAK cells. METHODS: A-LAK cells were obtained from peripheral blood mononuclear cells (PBMCs) of the patients with hepatocellular carcinoma (HCC) by using L-phenylalanine methyl ester (PME) to deplete immunosuppressive monocytes. Proliferative activity of SMMC7721 cell line after treatment with phenylacetate (PA) was observed. A-LAK cells were treated with the supernatant of SMMC7721 cells that had been pretreated with PA. The changes of proliferation, cytotoxicity and phenotype of A-LAK cells were investigated after cryopreservation. RESULTS: The expansion of A-LAK cells (96.79 +/- 69.10 folds on Day 14) was significantly higher than that of non-adherent LAK (NA-LAK) cells (22.77 +/- 13.20) as well as conventional LAK cells (4.64 +/- 0.91). PA significantly suppressed the growth of SMMC7721 cells, and the inhibitor ratio was 46%. The supernatant of cultured tumor cells intensively suppressed the proliferation and cytotoxicity of A-LAK cells, but the suppressive effect of the supernatant was previously decreased after treatment with PA. Impairments in proliferation and cytotoxicity of A-LAK cells immediately after thawing of cryopreservation and recovery after reincubation with IL-2 were observed. The cytotoxicity of thawed A-LAK cells on Day 5 was significantly higher than that of fresh A-LAK before freezing (54.8 +/- 10.2% vs 40.5 +/- 6.4%). No significant change in the percentage of lymphocyte subsets was identified in frozen A-LAK cells as compared with that in the fresh control cells. CONCLUSION: A-LAK cells can be simply prepared by using PME, and showed a synergistic anti-tumor effect with the combination of PA. Cryopreservation can increase the immunoactivities of A-LAK cells from the patients with hepatocellular carcinoma. PMID- 11925599 TI - Prediction of recurrence and prognosis in patients with hepatocellular carcinoma after resection by use of CLIP score. AB - AIM: The survival time of patients with hepatocellular carcinoma (HCC) after resection is hard to predict. Both residual liver function and tumor extension factors should be considered. A new scoring system has recently been proposed by the Cancer of the Liver Italian Program (CLIP). CLIP score was confirmed to be one of the best ways to stage patients with HCC. To our knowledge, however, the literature concerning the correlation between CLIP score and prognosis for patients with HCC after resection was not published. The aim of this study is to evaluate the recurrence and prognostic value of CLIP score for the patients with HCC after resection. METHODS: A retrospective survey was carried out in 174 patients undergoing resection of HCC from January 1986 to June 1998. Six patients who died in the hospital after operation and 11 patients with the recurrence of the disease were excluded at 1 month after hepatectomy. By the end of June 2001, 4 patients were lost and 153 patients with curative resection have been followed up for at least three years. Among 153 patients, 115 developed intrahepatic recurrence and 10 developed extrahepatic recurrence, whereas the other 28 remained free of recurrence. Recurrences were classified into early (< or =3 year) and late (>3 year) recurrence. The CLIP score included the parameters involved in the Child-Pugh stage (0-2), plus macroscopic tumor morphology (0-2), AFP levels (0-1), and the presence or absence of portal thrombosis (0-1). By contrast, portal vein thrombosis was defined as the presence of tumor emboli within vascular channel analyzed by microscopic examination in this study. Risk factors for recurrence and prognostic factors for survival in each group were analyzed by the chi-square test, the Kaplan-Meier estimation and the COX proportional hazards model respectively. RESULTS: The 1-, 3-, 5-, 7-,and 10-year disease-free survival rates after curative resection of HCC were 57.2%, 28.3%, 23.5%, 18.8%, and 17.8%, respectively. Median survival time was 28, 10, 4, and 5 mo for CLIP score 0, 1, 2, 3, and 4 to 5, respectively. Early and late recurrence developed in 109 patients and 16 patients respectively. By the chi-square test, tumor size, microsatellite, venous invasion, tumor type (uninodular, multinodular, massive), tumor extension (< or = or >50% of liver parenchyma replaced by tumor), TNM stage, CLIP score, and resection margin were the risk factors for early recurrence, whereas CLIP score and Child-Pugh stage were significant risk factors for late recurrence. In univariate survival analysis, Child-Pugh stages, resection margin, tumor size, microsatellite, venous invasion, tumor type, tumor extension, TNM stages, and CLIP score were associated with prognosis. The multivariate analysis by COX proportional hazards model showed that the independent predictive factors of survival were resection margins and TNM stages. CONCLUSION: CLIP score has displayed a unique superiority in predicting the tumor early and late recurrence and prognosis in the patients with HCC after resection. PMID- 11925600 TI - Mechanical properties of hepatocellular carcinoma cells. AB - AIM: To study the viscoelastic properties of human hepatocytes and hepatocellular carcinoma (HCC) cells under cytoskeletal perturbation, and to further to study the viscoelastic properties and the adhesive properties of mouse hepatoma cells (HTC) in different cell cycle. METHODS: Micropipette aspiration technique was adopted to measure viscoelastic coefficients and adhesion force to collagen coated surface of the cells. Three kinds of cytoskeleton perturbing agents, colchicines (Col), cytochalasin D (CD) and vinblastine (VBL), were used to treat HCC cells and hepatocytes and the effects of these treatment on cell viscoelastic coefficients were investigated. The experimental results were analyzed with a three-element standard linear solid. Further, the viscoelastic properties of HTC cells and the adhesion force of different cycle HTC cells were also investigated. The synchronous G(1) and S phase cells were achieved through thymine-2 desoryriboside and colchicines sequential blockage method and thymine-2 desoryriboside blockage method respectively. RESULTS: The elastic coefficients, but not viscous coefficient of HCC cells (K(1)=103.6+/-12.6N.m(-2), K(2)=42.5 +/ 10.4N.m(-2), mu=4.5 +/- 1.9Pa.s), were significantly higher than the corresponding value for hepatocytes (K(1)=87.5 +/- 12.1N.m(-2), K(2)=33.3+/ 10.3N.m(-2), mu=5.9+/-3.0Pa.s, P<0.01). Upon treatment with CD, the viscoelastic coefficients of both hepatocytes and HCC cells decreased consistently, with magnitudes for the decrease in elastic coefficients of HCC cells (K(1): 68.7 N.m( 2) to 81.7N.m(-2), 66.3% to 78.9%; K(2): 34.5N.m(-2) to 37.1N.m(-2), 81.2% to 87.3%, P<0.001) larger than those for normal hepatocytes (K(1): 42.6N.m(-2) to 49.8N.m(-2), 48.7% to 56.9%; K(2): 17.2N.m(-2) to 20.4N.m(-2), 51.7% to 61.3%, P<0.001). There was a little decrease in the viscous coefficient of HCC cells (2.0 to 3.4Pa.s, 44.4 to 75.6%, P<0.001) than that for hepatocytes (3.0 to 3.9Pa.s, 50.8 to 66.1% P<0.001). Upon treatment with Col and VBL, the elastic coefficients of hepatocytes generally increased or tended to increase while those of HCC cells decreased. HTC cells with 72.1% of G(1) phase and 98.9% of S phase were achieved and high K(1), K(2) value and low mu value were the general characteristics of HTC cells. G(1) phase cells had higher K(1) value and lower mu value than S phase cells had, and G(1) phase HTC cells had stronger adhesive forces ((275.9 +/- 232.8) x 10(-10)N) than S phase cells ((161.2 +/- 120.4) x 10( 10)N, P<0.001). CONCLUSION: The difference in both the pattern and the magnitude of the effect of cytoskeletal perturbing agent on the viscoelastic properties between HCC cells and hepatocytes may reflect differences in the state of the cytoskeleton structure and function and in the sensitivity to perturbing agent treatment between these two types of cells. Change in the viscoelastic properties of cancer cells may affect significantly tumor cell invasion and metastasis as well as interactions between tumor cells and their micro-mechanical environments. PMID- 11925601 TI - Study of apoptosis in human liver cancers. AB - AIM: To investigate the action of apoptosis in occurrence of liver carcinomas in vivo and the biological effect of Solanum lyratum Thumb on BEL-7404 cell line inducing apoptosis in vivo. METHODS: The apoptosis in the liver carcinoma was detected with terminal deoxynucl neotidyl transferase mediated dUTP nick end labelling (TUNEL); the cancer cells cultured in DMED medium were treated with extract of Solanum lyratum Thumb and observed under microscope, and their DNA was assayed by gel electrophoresis. RESULTS: In vivo apoptotic cells in the cancer adjacent tissues inceased; in vivo treatment of liver cancers with extract of Solanum lyratum Thumb could induce the cells to manifest a typical apoptotic morphology. Their DNA was fractured and a characteristic ladder pattern could be found using electrophoresis. CONCLUSION: In vivo the apoptosis of carcinomas was lower; maybe the cells divided quickly and then the cancers occurred. In the cancer adjacent tissues,the apoptosis pricked up, and in vivo Solanum lyratum Thumb could induce the apoptosis of BEL-7404 cells. PMID- 11925602 TI - Inhibitory effect of endostatin expressed by human liver carcinoma SMMC7721 on endothelial cell proliferation in vitro. AB - AIM: To construct a stable transfectant of human liver carcinoma cell line SMMC7721 that could secret human endostatin and to explore the effect of human endostatin expressed by the transfectant on endothelial cell proliferation. METHODS: Recombinant retroviral plasmid pLncx-Endo containing the cDNA for human endostatin gene together with rat albumin signal peptide was engineered and transferred into SMMC7721 cell by lipofectamine. After selection with G418, endostatin-transfected SMMC7721 cells were chosen and expanded. Immunohistochemical staining and Western blot were used to detect the expression of human endostatin in transfected SMMC7721 cells and its medium. The conditioned medium of endostatin-transfected and control SMMC7721 cells were collected to cultivate with human umbilical vein endothelial cells for 72 hours. The inhibitory effect of endostatin, expressed by transfected SMMC7721 cells, on endothelial proliferation in vitro was observed by using MTT assay. RESULTS: A 550 bp specific fragment of endostatin gene was detected from the PCR product of endostatin-transfected SMMC7721 cells. Immunohistochemistry and Western blot analysis confirmed the expression and secretion of foreign human endostatin protein by endostatin-transfected SMMC7721 cells. In vitro endothelial proliferation assay showed that 72 hours after cultivation with human umbilical vein endothelial cells, the optical density (OD) in group using the medium from endostatin-transfected SMMC7721 cells was 0.51 +/- 0.06, lower than that from RPMI 1640 group (0.98 +/- 0.09) or that from control plasmid pLncx-transfected SMMC7721 cells (0.88 +/- 0.11). The inhibitory rate for medium from endostatin transfected SMMC7721 cells was 48%, significantly higher than that from empty plasmid pLncx-transfected SMMC7721 cells (10.2%, P<0.01). CONCLUSION: Human endostatin can be stably expressed by SMMC7721 cell transferred with human endostatin gene and its product can significantly inhibit the proliferation of human umbilical vein endothelial cell in vitro. PMID- 11925603 TI - Expression and identification of recombinant soluble single-chain variable fragment of monoclonal antibody MC3. AB - AIM: To generate soluble single chain variable fragments (ScFv) of monoclonal antibody MC3 recognizing colorectal and gastric carcinomas. METHODS: mRNA was isolated from the hybridoma cell line producing MC3 and the DNAs encoding variable domains of heavy and light chains (VH and VL) of the antibody were amplified separately by RT-PCR and assembled into ScFv DNA with a linker DNA. The ScFv DNA was ligated into the phagemid vector pCANTAB5E and the ligated sample was transformed into E.coli TG1. The transformed cells were infected with M13KO7 helper phage to yield recombinant phages. After two rounds of panning with gastric carcinoma cell line AGS highly expressing MC3-binding antigen, the phage clones displaying ScFv fragments of the antibody were selected by ELISA. 4 phage clones showing strong signal in ELISA were used to infect E.coli HB2151 to express soluble ScFvs. The soluble ScFvs were identified by Dot blot and Western blot, and their antigen-binding activity was assayed by ELISA. The VH and VL DNAs of the ScFv DNA derived from phage clone 19 were sequenced. RESULTS: The VH,VL and ScFv DNAs were about 340 bp, 320 bp and 750 bp respectively. After two rounds of panning to the recombinant phages, 18 antigen-positive phage clones were selected from 30 preselected phage clones by ELISA. All the soluble ScFvs derived from the 4 out of the 18 antigen-positive phage clones were about M(r)32000 and concentrated in periplasmatic space under the given culture condition. The soluble ScFvs could bind the antigen, and they shared the same binding site with MC3. The sequences of the VH and VL DNAs of the MC3 ScFv showed that the variable antibody genes belonged to the IgG1 subgroup,kappa-type. CONCLUSION: The soluble ScFv of MC3 is successfully produced, which not only provides a possible novel targeting vehicle for in vivo and in vitro study on associated cancers, but also offers the antibody a stable genetic source. PMID- 11925604 TI - Report of 16 kindreds and one kindred with hMLH1 germline mutation. AB - AIM: To analyze the diagnosis and treatment of 16 hereditary nonpolyposis colorectal cancer (HNPCC) kindreds, and to report the first kindred with hMLH1 germline mutation in Mainland China. METHODS: The diagnosis, treatment and follow up study of 16 HNPCC kindreds were retrospectively reviewed. Data concerning site of the malignant tumor, age at the diagnosis, history of synchronous and/or metachronous cancer, and histopathology of tumors were recorded. All treatments had won formal consent. PCR and SSCP were used to screen the coding region of hMLH1 and hMSH2 genes. Variant bands were sequenced by a 377 DNA sequencer. RESULTS: Among sixteen kindreds, sixty-eight patients had a mean age of 50.8 years, including twenty-one multiple cancer patients and forty-six colorectal cancer patients (metachronous colorectal cancers in sixteen). A total of one hundred and one malignant neoplasms were found in these sixty-eight patients, including 50 colonic, 17 rectal, 11 gastric, 7 endometrial, and 4 esophageal cancers. 39.5% colorectal patients had metachronous cancers within ten years who needed reoperations. A germline G265T nonsense mutation was found in the third exon of hMLH1, resulting in a stop codon and truncated protein. Three phenotypically normal family members were also found to carry the mutated gene. CONCLUSION: HNPCC is a typical auto-dominant hereditary disease, the main characteristics include early onset and frequency of cancers; predominance of colorectal, especially right-sided colon cancers; frequency of multiple primary cancers (especially colorectal cancers). Segmental resection for colorectal cancers is not eligible for colorectal cancer patient in HNPCC kindreds. Intensive follow-up is essential for all patients and possible gene carriers. The first HNPCC kindred with hMLH1 gene germline mutation was identified in Mainland China, and three phenotypically normal family members were found to be carriers of the mutated gene. The G265T germline (nonsense) mutation in the third exon of hMLH1 found here had not been reported previously in the literature. PMID- 11925605 TI - Colonoscopic screening and follow-up for colorectal cancer in the elderly. AB - AIM: To improve the prevention and treatment of senile patients with colorectal cancer by evaluating the importance of colonoscopy in clinical screening and follow-up. METHODS: Clinical screening of colonoscopy was performed for 2196 patients aged 60-90 years old according to the protocol,and 1740 of them (79.2%) were followed-up. RESULTS: Colorectal cancer was found in 52 patients, and the detectable rate was 2.4%. Among them, 19 were diagnosed as early colorectal cancer, accounting for 36.5% of the detected colorectal cancer. Among the followed-up patients, early colorectal cancer was found in 9, accounting for 45.0% of the detected colorectal cancer. The resectable rate and 5 years survival rate of colorectal cancer were 97.7% and 80.9% respectively. The incidence of complication was 0.05%, and the successful rate of cecum intubation was 98.9%. CONCLUSION: Colonoscopic screening and follow-up of the elderly for colorectal cancer and pre-cancerous lesion (adenomatoid polyp) can increase the detectable rate of early colorectal cancer and improve its prevention and treatment. PMID- 11925606 TI - Specific CEA-producing colorectal carcinoma cell killing with recombinant adenoviral vector containing cytosine deaminase gene. AB - AIM: To kill CEA positive colorectal carcinoma cells specifically using the E coli cytosine deaminase (CD) suicide gene, a new replication-deficient recombinant adenoviral vector was constructed in which CD gene was controlled under CEA promoter and its in vitro cytotoxic effects were evaluated. METHODS: Shuttle plasmid containing CD gene and regulatory sequence of the CEA gene was constructed and recombined with the right arm of adenovirus genome DNA in 293 cell strain. Dot blotting and PCR were used to identify positive plaques. The purification of adenovirus was performed with ultra-concentration in CsCl step gradients and the titration was measured with plaque formation assay. Cytotoxic effects were assayed with MTT method, The fifty percent inhibition concentration (IC(50)) of 5-FC was calculated using a curve-fitting parameter. The human colorectal carcinoma cell line, which was CEA-producing, and the CEA-nonproducing Hela cell line were applied in cytological tests. An established recombinant adenovirus vector AdCMVCD, in which the CD gene was controlled under CMV promoter, was used as virus control. Quantitative results were expressed as the mean +/- SD of the mean. Statistical analysis was performed using ANOVA test. RESULTS: The desired recombinant adenovirus vector was named AdCEACD. The results of dot blotting and PCR showed that the recombinant adenovirus contained CEA promoter and CD gene. Virus titer was about 5.0 X 10(14)pfu/L(-1) after purification. The CEA-producing Lovo cells were sensitive to 5-FC and had the same cytotoxic effect after infection with AdCEACD and AdCMVCD (The IC(50) values of 5-FC in parent Lovo cells, Lovo cells infected with 100 M.O.I AdCEACD and Lovo cells infected with 10 M.O.I AdCMVCD were >15000, 216.5+/-38.1 and 128.8+/-25.4 micromol.L(-1), P<0.001, respectively), and the cytotoxicity of 5-FC increased accordingly when the m.o.i of adenoviruses were enhanced (The value of IC(50) of 5-FC was reduced to 27.9+/-4.2 micromol.L(-1) in 1000 M.O.I AdCEACD infected Lovo cells and 24.8+/-7.1 micromol.L(-1) in 100 M.O.I AdCMVCD infected Lovo cells, P<0.05, P<0.01, respectively). The CEA-nonproducing Hela cells had no effect after infection with AdCEACD, but Hela cells had the cytotoxic sensitivity to 5 FC after infection with AdCMVCD (The IC(50) of 5-FC in parent Hele cells and Hela cells infected with AdCMVCD at 10 M.O.I was >15000 and 214.5+/-31.3 micromol.L( 1), P<0.001). AdCEACD/5-FC system also had bystander effect, and the viability was about 30 percent when the proportion of transfected cells was only 10 percent. CONCLUSION: The recombinant adenovirus vector AdCEACD has the character of cell type-specific gene delivery. The AdCEACD/5-FC system may become a new, potent and specific approach for the gene therapy of CEA-positive neoplasms, especially colon carcinoma. PMID- 11925608 TI - Methodologic research on TIMP-1, TIMP-2 detection as a new diagnostic index for hepatic fibrosis and its significance. AB - AIM: To set up a new method to detect tissue inhibitors of metalloproteinase-1 and -2(TIMP-1 and TIMP-2) in sera of patients with hepatic cirrhosis, and to investigate the expression and location of TIMP-1 and TIMP-2 in liver tissue of patients with hepatic cirrhosis, and the correlation between TIMPs in liver and those in sera so as to discuss whether TIMPs can be used as a diagnosis index of hepatic fibrosis. METHODS: The monoclonal antibodies (McAbs) of TIMP-1 and TIMP-2 were used to sensitize erythrocytes, and solid-phase absorption to sensitized erythrocytes (SPASE) was used to detect TIMP-1 and TIMP-2 in the sera of patients with hepatic cirrhosis. Meanwhile, with the method of in situ hybridization and immunohistochemistry, we studied the mRNA expression and antigen location of TIMP 1 and TIMP-2 in the livers of 40 hepatic cirrhosis patients with pathologic diagnosis. RESULTS: With SPASE, they were 16.4% higher in the acute hepatitis group, 33.3% higher in the chronic hepatitis group, and the positive rates were 73.6% and 61.2% respectively in sera of hepatic cirrhosis patients, which were remarkably higher than those in chronic hepatitis and acute hepatitis group (P<0.001). In 40 samples of hepatic cirrhosis tissues, all of them showed positive expression of TIMP-1 and TIMP-2 mRNA detected with immunohistochemistry or in situ hybridization (positive rate was 100%). Expression of TIMPs in different degrees could be found in liver tissue with cirrhosis. TIMPs were located in cytoplasm of liver cells of patients with hepatic cirrhosis. There was a significant correlation between serum TIMPs level and liver TIMPs level. CONCLUSION: SPASE is a useful method to detect the TIMP-1 and TIMP-2 in sera of patients with hepatic cirrhosis, and TIMP-1 and TIMP-2 can be considered as a useful diagnostic index of hepatic fibrosis, especially TIMP-1. PMID- 11925607 TI - Detection of anti-preS1 antibodies for recovery of hepatitis B patients by immunoassay. AB - AIM: To establish a convenient immunoassay method based on recombinant antigen preS1(21-119aa) to detect anti-preS1 antibodies and evaluate the clinical significance of antibodies in hepatitis B. METHODS: The expression plasmid pET 28a-preS1 was constructed, and a large quantity of preS1(21-119aa) fragment of the large HBsAg protein was obtained. The preS1 fragment purified by Ni(2+)-IDA affinity chromatography was used as coated antigen to establish the indirect ELISA based on streptavidin-biotin system for detection of the anti-preS1 antibodies in sera from HBV-infected patients. For follow-up study, serial sera were collected during the clinical course of 21 HBV-infected patients and anti preS1 antibodies, preS1 antigen, HBV-DNA and other serological HBV markers were analyzed. RESULTS: preS1(21-119aa) fragment was highly expressed from the plasmid pET-28a-preS1 in a soluble form in E.Coli (30mg.L(-1)), and easily purified to high purity over 90% by one step of Ni(2+)-IDA-sepharose 6B affinity chromatography. The purity and antigenicity of the purified preS1(21-119aa) protein was determined by 150g.L(-1) SDS-PAGE, Western blot and a direct ELISA. Recombinant preS1(21-119aa) protein was successfully applied in the immunoassay which could sensitively detect the anti-preS1 antibodies in serum specimens of acute or chronic hepatitis B patients. Results showed that more than half of 19 acute hepatitis B patients produced anti-preS1 antibodies during recovery of the disease, however, the response was only found in a few of chronic patients. In the clinical follow-up study of 11 patients with anti-preS1 positive serological profile, HBsAg and HBV-DNA clearance occurred in 6 of 10 acute hepatitis B patients in 5-6 months, and seroconversion of HBeAg and disappearance of HBV-DNA occurred in 1 chronic patients treated with lavumidine, a antiviral agent. CONCLUSION: The high-purity preS1(21-119aa) coated antigen was successfully prepared by gene expression and affinity chromatography. Using this antigen, a conveniently detective system of anti-preS1 antibodies in sera was established. Preliminarily clinical trial the occurrence of anti-preS1 antibodies in acute hepatitis B patients suggests the clearance of HBV from serum in a short-term time, and anti-preS1 positive in chronic patients means health improvement or recovery from the disease. PMID- 11925609 TI - Clinicopathological study on TTV infection in hepatitis of unknown etiology. AB - AIM: To investigate the state of infection, replication site, pathogenicity and clinical significance of transfusion transmitted virus (TTV) in patients with hepatitis, especially in patients of unknown etiology. METHODS: Liver tissues taken from 136 cases of non-A non-G hepatitis were tested for TT virus antigen and nucleic acid by in situ hybridization (ISH) and nested-polymerase chain reaction (PCR). Among them, TT virus genome and its complemental strand were also detected in 24 cases of autopsy liver and extrahepatic tissues with ISH. Meanwhile, TTV DNA was detected in the sera of 187 hepatitis patients by nested PCR. The pathological and clinical data of the cases infected with TTV only were analyzed. RESULTS: In liver, the total positive rate of TTV DNA was 32.4% and the positive signals were located in the nuclei of hepatocytes. In serus, TTV DNA was detected in 21.4% cases of hepatitis A-G, 34.4% of non-A non-G hepatitis and 15% of healthy donors. The correspondence rate of TTV DNA detection between liver tissue with ISH and sera with PCR was 63.2% and 89.3% in the same liver tissues by ISH and by PCR, respectively. Using double-strand probes and single-strand probes designed to detect TTV genome, the correspondence rate of TTV DNA detected in liver and extrahepatic tissues was 85.7%. Using single-strand probes, TTV genome could be detected in liver and extrahepatic tissues by PCR, but its complemental strands (replication strands) could be observed only in livers. The liver function of most cases infected with TTV alone was abnormal and the liver tissues had different pathological damage such as ballooning, acidophilia degeneration, formation of apoptosis bodies and focus of necrosis, but the inflammation in the lobule and portal area was mild. CONCLUSION: The positive rate of TTV DNA among cases of hepatitis was higher than that of donors, especially in patients with non-A non-G hepatitis, but most of them were coinfected with other hepatitis viruses. TTV can infect not only hepatocytes, but also extrahepatic tissues. However, the chief replication place may be liver. The infection of TTV may have some pathogenicity. Although the pathogenicity is comparatively weak, it can still damage the liver tissues. The lesions in acute hepatitis (AH) and chronic hepatitis (CH) are mild, but in severe hepatitis (SH), it can be very serious and cause liver function failure, therefore, we should pay more attention to TTV when studying the possible pathogens of so-called "liver hepatitis of unknown etiology". PMID- 11925610 TI - Construction and characterization of an experimental ISCOMS-based hepatitis B polypeptide vaccine. AB - AIM: To characterize the biochemical and immunological properties of an experimental ISCOMS vaccine prepared from a novel therapeutic polypeptide based on T cell epitopes of HBsAg, and a heptatis B-ISCOMS was prepared and investigated. METHODS: An immunostimulating complexes(ISCOMS)-based vaccine containing a novel therapeutic hepatitis B polypeptide was prepared by dialysis method, and its formation was visualized by electron microscopy and biochemically verified by SDS-polyacrylamide gel electrophoresis. Amount of the peptide within ISCOMS was determined by Bradford assay, and specific CTL response was detected by ELISPOT assay. RESULTS: Typical cage-like structures of submicroparticle with a diameter of about 40nm were observed by electron microscopy. Results from Bradford assay showed that the level of peptide incorporation was about 0.33g.L( 1). At the paralleled position close to the sixth band of the molecular weight marker(3480kDa) a clear band was shown in SDS-PAGE analysis, indicating successful incorporation of polypeptide into ISCOMS. It is suggested that ISCOMS delivery system could efficiently improve the immunogenicity of polypeptide and elicit specific immune responses in vivo by the results of ELISPOT assay, which showed that IFN-gamma producing cells(specific CTL responses) were increased(spots of ISCOMS-treated group: 47+/-5, n =3; control group: 5+/-2, n =3). CONCLUSION: ISCOMS-based hepatitis B polypeptide vaccine is successfully constructed and it induces a higher CTL response compared with short polypeptides vaccine in vivo. PMID- 11925613 TI - Detection of H.pylori DNA in gastric epithelial cells by in situ hybridization. AB - AIM: To investigate the presence of H.pylori DNA within gastric epithelial cells in patients with H.pylori infection and its possible carcinogenic mechanism. METHODS: Total 112 patients, with pathologically confirmed chronic superficial gastritis, chronic atrophic gastritis, intestinal metaplasia, atypical hyperplasia or gastric cancer were studied. Among them, 28 were H.pylori negative and 84 H.pylori positive. H.pylori DNA in gastric epithelial cells was detected by GenPoint catalyzed signal amplification system for in situ hybridization. RESULTS: In the H.pylori positive group, zero out of 24 chronic superficial gastritis (0.0%), four out of 25 precancerous changes (16.0%) and thirteen out of 35 gastric cancers (37.1%) showed H.pylori DNA in the nucleus of gastric epithelial cells, the positive rates of H.pylori DNA in the nucleus of gastric epithelial cells were progressively increased in chronic superficial gastritis, precancerous changes and gastric cancer groups (chi(2)=12.56, P=0.002); One out of 24 chronic superficial gastritis (4.2%), eleven out of 25 precancerous changes (44.0%) and thirteen out of 35 gastric cancers (37.1%) showed H.pylori DNA in the cytoplasm of gastric epithelial cells (chi(2)=10.86, P=0.004). In the H.pylori negative group, only one patient with gastric cancer was found H.pylori DNA in the nucleus of gastric epithelial cells; Only two patients, one patient with precancerous changes and another with gastric cancer, showed H.pylori DNA in the cytoplasm of gastric epithelial cells. Furthermore, H.pylori DNA must have been in the cytoplasm as long as it existed in the nucleus of gastric epithelial cells. CONCLUSION: H.pylori DNA exists both in the nucleus and the cytoplasm of gastric epithelial cells in patients with H.pylori infections. The pathological progression from chronic superficial gastritis, precancerous changes to gastric cancer is associated with higher positive rates of H.pylori DNA presence in the nucleus of gastric epithelial cells. PMID- 11925611 TI - mIL-2R, T cell subsets & hepatitis C. AB - AIM: To study the levels of membrane interleukin-2 receptor (mIL-2R) and T cell subsets in peripheral blood mononuclear cells (PBMC) from patients with hepatitis C and their role in the pathogenesis of hepatitis C. METHODS: The levels of mIL 2R and T cells subsets in PBMC were detected by biotin- streptatividin (BSA) technique before and after stimulation with PHA in 203 patients with hepatitis C with HCV-RNA(+), anti-HCV(+), anti-HCV(-). RESULTS: The total expressive levels of mIL-2R before and after stimulation with PHA(0.03+/-0.01, 0.03+/-0.02, 0.04+/ 0.02, 0.36+/-0.03), and T cell subsets in PBMC (0.62+/-0.06, 0.37+/-0.05, 0.35+/ 0.07) were all lower in patients with hepatitis C than those in normal controls (0.66+/-0.07, 0.41+/-0.06, 0.31+/-0.05, P<0.01). Among the patients, the levels of mIL-2R were lower in silence than those in situation of PHA inducting (P<0.01). However, the levels of mIL-2R were similar in acute hepatitis C to that in chronic hepatitis C (P>0.05). The levels of CD(3)(+), CD(4)(+), CD(4)(+)/CD(8)(+) were lower and CD(8)(+) was higher in patients with acute and chronic hepatitis C with anti-HCV(+) than those in normal controls (0.62+/-0.06, 0.37+/-0.05, 0.35+/-0.07, 1.18+/-0.30, 0.61+/-0.07, 0.37+/-0.05, 1.39+/-0.33, 0.31+/-0.05, P<0.05-P<0.01). CONCLUSION: The cellular immunity is obviously changed in patients with hepatitis C. The levels of mIL-2R and activation of T cells are closely associated with chronicity of hepatitis C. PMID- 11925612 TI - Modalities of testing Helicobacter pylori in patients with nonmalignant bile duct diseases. AB - AIM: This paper describes the procedure of detection of Helicobacter pylori (H. pylori) in bile specimens in patients suffering from benign diseases of biliary ducts (lithiasis with/without nonspecific cholangitis). METHODS: The group of 72 patients entering the study consisted of 32 male and 40 female (45% and 55%, respectively). Bile was obtained during ERCP in 68 patients, and during cholecystectomy in 4 patients. A fast urease test (FUT) to determine the existence of H. pylori in gastric mucosa was carried out for all the patients during the endoscopic examination. The existence of genetic material of H. pylori was determined by detection of ureA gene by the method of nested PCR. The results of this reaction were shown by electrophoresis on 10g.L(-1) agarose gel in a band of 256bp. RESULTS: The majority of the patients included in our study had biliary lithiasis without signs of cholangitis (48 patients, 67%), whereas other patients were complicated by cholangitis (17 patients, 24%). Seven patients (9%) had normal ERCP, forming thus the control group. In the group of patients with lithiasis 26 patients (54.2%) had positive PCR of H. pylori in bile and among the patients with associated cholangitis positive PCR was detected in 9 patients (52.9%). Among the seven patients with normal ERCP only one (14%) had positive PCR of H. pylori. A high percentage of H. pylori infection of gastric mucosa was observed (57 patients, 79%). It was also observed that its slightly higher positivity was in the patients with distinct bile pathology: 81% FUT positive patients in the group with choledocholithiasis alone and 76% in the group with choledocholithiasis associated with cholangitis. Seventy-one percent of the patients with regular findings had positive FUT. CONCLUSION: The prevalence of H. pylori infection both in bile and in gastric mucosa in patients with benign diseases of biliary ducts does not show a statistically significant difference in relation to the prevalence of the same with the patients with normal ERCP. The existence of H. pylori infection possibly does not play a role in pathogenesis of benign biliary diseases. PMID- 11925614 TI - A study of recombinant protective H.pylori antigens. AB - AIM: To construct a recombinant vector which can express M (r)26000 outer membrane protein (OMP) from Helicobacter pylori (Hp), and to obtain the vaccine protecting against Hp infection and a diagnostic reagent kit quickly detecting Hp infection. METHODS: The gene encoding the structural M(r)26000 outer membrane protein of Hp was amplified from Hp chromosomal DNA by PCR, and inserted in the prokaryotic expression vector pET32a (+), which was transformed into the Top10 E. coli strain. Recombinant vector was selected, identified and transformed into BL 21(DE3) E. coli strain. The recombinant fusion proteins were expressed. The antigenicity of recombinant protein was studied by ELISA or immunoblotting and immunized Balb/c mice. RESULTS: The gene of M(r)26000 OMP was amplified to be 594 base pairs, 1.1% of the cloned genes was mutated and 1.51% of amino acid residues was changed, but there was homogeneity between them. The recombinant fusion protein encoded objective polypeptides of 198 amino acid residues, corresponding to calculated molecular masses of M (r)26000. The level of soluble expression products was about 38.96% of the total cell protein. After purification by Ni-NTA agarose resin columniation, the purity of objective protein became about 90%. The ELISA results showed that recombinant fusion protein could be recognized by patient serum infected with Hp and rabbit serum immunized with the recombinant protein. Furthermore,Balb/c mice immunized with the recombinant protein were protected against H.pylori infection. CONCLUSION: M (r)26000 OMP may be a candidate vaccine preventing Hp infection. PMID- 11925615 TI - Morphological properties and residual strain along the small intestine in rats. AB - AIM: Residual stress and strain are important for gastrointestinal function and relate to the geometric configuration, the loading conditions and the zero-stress state of the gastrointestinal tract. The purpose of this project is to provide morphometric data and residual strains for the rat small intestine ( n =11). METHODS: To approach the no-load state, the intestine was surgically excised, transferred to an organ bath and cut transversely into short ring-shaped segments. Each ring was cut radially for obtaining the zero-stress state. The residual stress can be characterised by an opening angle. The strain difference between the zero-stress state and the no-load state is called residual strain. RESULTS: Large morphometric variations were found along the small intestine. The wall thickness was highest in the proximal duodenum and decreased in distal direction along the axis of the small intestine (P<0.001). The circumferential length of the inner and outer surfaces decreased rapidly along the length of duodenum by 30-50% (P<0.001). The wall area and lumen area showed a similar pattern (P<0.001). In zero-stress state the rings always opened up after making the cut. The experiments resulted in larger inner circumferential length and smaller outer circumferential length when compared to the no-load state. The wall thickness and wall area did not differ between the no-load and zero-stress state. The opening angle and tangent rotation angle increased along the length of the duodenum and had its highest value 30% down the intestine. Further down the intestine it decreased again (P<0.001). The serosal residual strain was tensile with the highest value close to the ligament of Treitz (P<0.001). The mucosal residual strain was compressive in all segments of the small intestine with average values between -0.25 and -0.4 and with the lowest values close to the ligament of Treitz (P<0.001). CONCLUSION: Axial variation in morphometric properties and residual strains were found in the small intestine. Existence of large residual strains indicates that the zero-stress state must be considered in future biomechanical studies in the gastrointestinal tract. PMID- 11925616 TI - Cloning of cytochrome P-450 2C9 cDNA from human liver and its expression in CHL cells. AB - AIM: Using bacterial, yeast, or mammalian cell expressing a human drug metabolism enzyme would seem good way to study drug metabolism-related problems. Human cytochrome P-450 2C9(CYP2C9) is a polymorphic enzyme responsible for the metabolism of a large number of clinically important drugs. It ranks among the most important drug metabolizing enzymes in humans. In order to provide a sufficient amount of the enzyme for drug metabolic research, the CYP2C9 cDNA was cloned and expressed stably in CHL cells. METHODS: After extraction of total RNA from human liver tissue, the human CYP2C9 cDNA was amplified with reverse transcription-polymerase chain reaction (RT-PCR), and cloned into cloning vector pGEM-T. The cDNA fragment was identified by DNA sequencing and subcloned into a mammalian expression vector pREP9. A transgenic cell line was established by transfecting the recombinant vector of pREP9-CYP2C9 into CHL cells. The enzyme activity of CYP2C9 catalyzing oxidation of tolbutamide to hydroxy tolbutamide in S9 fraction of the cell was determined by high performance liquid chromatography(HPLC). RESULTS: The amino acid sequence predicted from the cDNA segment was identical to that of CYP2C9*1, the wild type CYP2C9. However, there were two base differences, i.e. 21T>C, 1146C>T, but the encoding amino acid sequence was the same, L7, P382. The S9 fraction of the established cell line metabolizes tolbutamide to hydroxy tolbutamide; tolbutamide hydroxylase activity was found to be 0.465 +/- 0.109 micromol.min(-1).g(-1) S9 protein or 8.62 +/- 2.02mol.min(-1).mol(-1) CYP, but was undetectable in parental CHL cell. CONCLUSION: The cDNA of human CYP2C9 was successfully cloned and a cell line of CHL- CYP2C9, efficiently expressing the protein of CYP2C9, was established. PMID- 11925617 TI - Infrared thermoimages display of body surface temperature reaction in experimental cholecystitis. AB - AIM: To display the thermoimages of the body surface in experimental cholecystitis, to observe the body surface temperature reaction in visceral disorders, and to study if the theory of body surface-viscera correlation is true and the mechanism of temperature changes along the meridians. METHODS: By injecting bacteria suspension into the stricture bile duct and gallbladder, 21 rabbits were prepared as acute pyogenic cholangiocholecystitis models, with another 8 rabbits prepared by the same process except without injection of bacteria suspension as control. The body surface infrared thermoimages were continuously observed on the hair shaven rabbit skin with AGA-782 thermovision 24h before, 1-11 d after and (2,3 wk) 4 wk after the operation with a total of over 10 records of thermoimages. RESULTS: Twelve cases out of 21 rabbits with cholecystitis revealed bi-lateral longitudinal high temperature lines in its trunk; with negative findings in the control group. The high-temperature line appeared on d1-d2, first in the right trunk, after the preparation of the model, about 7 d after the model preparation, the lines appeared at the left side too, persisting for 4 wk. The hyper-temperature line revealed 1.1-2.7 degrees higher than before the model preparation, 0.7-2.5 degrees higher than the surrounding skin. The length of the high temperature line might reach a half length of the body trunk, or as long as the whole body itself. CONCLUSION: The appearance of the longitudinal high temperature lines at the lateral aspects of the trunk in the experimental group is directly bound up with the experimental animals pyogenic cholecystitis, with its running course quite similar to that of the Gallbladder Channel of Foot Shaoyang, but different to the zones of hyperalgesia and site of referred pain in cholecystitis. PMID- 11925618 TI - Effect of L-NAME on nitric oxide and gastrointestinal motility alterations in cirrhotic rats. AB - AIM: To investigate the effect of L-NAME on nitric oxide and gastrointestinal motility alterations in cirrhotic rats. METHODS: Rats with cirrhosis induced by carbon tetrachloride were randomly divided into two groups, one n =13 receiving 0.5mg.kg(-1) per day of N(G)-nitro-L-arginine methyl ester (L-NAME), a nitric oxide synthase inhibitor, for 10 days, whereas the other group (n =13) and control (n =10) rats were administrated the same volume of 9g.L(-1) saline. Half gastric emptying time and 2h residual rate were measured by SPECT, using (99m)Tc DTPA-labeled barium sulfate as test meal. Gastrointestinal transition time was recorded simultaneously. Serum concentration of nitric oxide (NO) was determined by the kinetic cadmium reduction and colorimetric methods. Immunohistochemical SABC method was used to observe the expression and distribution of three types of nitric oxide synthase (NOS) isoforms in the rat gastrointestinal tract. Western blot was used to detect expression of gastrointestinal NOS isoforms. RESULTS: Half gastric emptying time and trans-gastrointestinal time were significantly prolonged(124.0 +/- 26.4 min; 33.7 +/- 8.9 min; 72.1 +/- 15.3 min; P<0.01), (12.4 +/- 0.5h; 9.5 +/- 0.3h; 8.2 +/- 0.8h; P<0.01), 2h residual rate was raised in cirrhotic rats than in controls and cirrhotic rats treated with L-NAME (54.9 +/- 7.6%,13.7 +/- 3.2%, 34.9 +/- 10.3%, P<0.01). Serum concentration of NO was significantly increased in cirrhotic rats than in the other groups (8.20 +/- 2.48) micromol.L(-1), (5.94 +/-1.07) micromol.L(-1) and control (5.66 +/- 1.60 micromol.L(-1), P<0.01. NOS staining intensities which were mainly located in the gastrointestinal tissues were markedly lower in cirrhotic rats than in the controls and cirrhotic rats after treated with L-NAME. CONCLUSION: Gastrointestinal motility was remarkably inhibited in cirrhotic rats, which could be alleviated by L-NAME. Nitric oxide may play an important role in the inhibition of gastrointestinal motility in cirrhotic rats. PMID- 11925619 TI - Regulating effect of Chinese herbal medicine on the peritoneal lymphatic stomata in enhancing ascites absorption of experimental hepatofibrotic mice. AB - AIM: To observe the regulatory effect of Chinese herbal medicine on peritoneal lymphatic stomata and its significance in treating ascites in liver fibrosis model mice. METHODS: Two Chinese herbal composite prescriptions were used separately to treat the carbon tetrachloride-induced mouse model of liver fibrosis. The histo-pathologic changes of the liver sections (HE and VG stainings) were observed. The peritoneal lymphatic stomata was detected by scanning electron microscopy and computer image processing. The changes of urinary volume and sodium ion concentration were measured. RESULTS: In the model group, lots of fibrous tissue formed in liver and extended into the hepatic lobules to separate them incompletely. In the treated and prevention groups, the histo-pathologic changes of liver was rather milder, only showed much less fibrous tissue proliferation in the hepatic lobules. The peritoneal lymphatic stomata enlarged with increased density in the experimental groups (diameter: PA, 3.07 +/- 0.69 microm; PB, 2.82 +/- 0.37 microm; TA, 3.25 +/- 0.82 microm and TB, 2.82 +/- 0.56 microm; density: PA, 7.11 +/- 1.90 stomata.1000 microm(-2); PB, 8.76 +/- 1.45 stomata.1000 microm(-2); TA, 6.55 +/- 1.44 stomata.1000 microm( 2)and TB, 8.76+/-1.79 stomata.1000 microm(-2)), as compared with the model group (diameter: 2.00+/-0.52 microm density: 4.45+/-1.05 stomata.1000 microm(-2)). After treatment, the urinary volume and sodium ion excretion increased in the experimental groups (PA, 231.28+/-41.09 mmol.L(-1); PB, 171.69 +/- 27.48 mmol.L( 1) and TA, 231.44 +/- 34.12 mmol.L(-1)), which were significantly different with those in the model group (129.33 +/- 36.75 mmol.L(-1)). CONCLUSION: Chinese herbal medicine has marked effects in alleviating liver fibrosis, regulating peritoneal lymphatic stomata, improving the drainage of ascites from peritoneal cavity and causing increase of urinary volume and sodium ion excretion to reduce the water and sodium retention, and thus have favorable therapeutic effect in treating ascites. PMID- 11925620 TI - Effects of progesterone on gastric emptying and intestinal transit in male rats. AB - AIM: To study the dose-dependent of progesterone (P) effect and the interaction between the oxytocin (OT) and P on gastrointestinal motility. METHODS: In order to monitor the gastric emptying and intestinal transit, the SD male rats were intubated via a catheter with normal saline (3 ml/kg) containing Na(2)(51)CrO(4) (0.5 microCi/ml) and 10% charcoal. OT was dissolved into normal saline and P was dissolved into 75% alcohol. RESULTS: Low does of P (1 mg/kg, i.p.) enhanced the gastric emptying (75+/-3%, P<0.05) and high dose of P (5 mg/kg, i.p.) inhibit it (42+/-11.2%, P<0.01). P (1 mg/kg) increased the intestinal transit (4.2+/-0.3, P<0.05) while the higher dose (10-20 mg/kg) had no effect. OT (0.8 mg/kg, i.p.) inhibited the gastric emptying (23.5+/-9.8%, P<0.01). The inhibitory effects of P(20 mg/kg) (32+/-9.7%, P<0.05) and OT (0.8 mg/kg) on gastric emptying enhanced each other when the two chemicals were administrated simultaneously (17+/-9.4%, P<0.01). CONCLUSION: Low dose of P increased GI motility while high dose of P decreased it. During the later period of pregnancy, elevated plasma level of OT may also participate in the gastrointestinal inhibition. PMID- 11925621 TI - Liver sinusoidal endothelial cell injury by neutrophils in rats with acute obstructive cholangitis. AB - AIM: The objective of this study is to elucidate the potential role of poly morphonuclear neutrophils (PMN) in the development of such a sinusoidal endothelial cell (SEC) injury during early acute obstructive cholangitis (AOC) in rats. METHODS: Twenty one Wistar rats were divided into three groups: the AOC group, the bile duct ligated group (BDL group), and the sham operation group (SO group). The common bile duct (CBD) of rats in AOC group was dually ligated and 0.2 ml of the E. Coli O(111) B(4) (5 X 10(9)cfu/ml) suspension was injected into the upper segment, in BDL group, only the CBD was ligated and in SO group, neither injection of E. Coli suspension nor CBD ligation was done, but the same operative procedure. Such group consisted of seven rats, all animals were killed 6h after the operation. Morphological changes of the liver were observed under light and electron microscope. Expression of intercellular adhesion molecule-1 (ICAM-1) mRNA in hepatic tissue was determined with reverse transcription polymerase chain reaction (RT-PCR). The serum levels of alanine aminotransferase (ALT) were determined with an autoanalyger and cytokine-induced neutrophil chemoattractant (CINC) was determined by enzyme-linked immunosorbent assay (ELISA). RESULTS: Neutrophils was accumulated in the hepatic sinusoids and sinusoidal endothelial cell injury existed in AOC group. In contrast, in rats of BDL group, all the features of SEC damage were greatly reduced. Expression of ICAM-1 mRNA in hepatic tissue in three groups were 7.54 +/- 0.82, 2.87 +/- 0.34, and 1.01 +/- 0.12, respectively. There were significant differences among three groups (P<0.05). The serum CINC levels in the three groups were 188 +/- 21 ng.L( 1), 94+/-11 ng.L(-1), and 57+/-8 ng.L(-1), respectively. There were also significant differences among the three groups (P<0.05). Activity of the serum ALT was 917 +/- 167 nkat.L(-1), 901 +/- 171 nkat.L(-1), and 908 +/- 164 nkat.L( 1), respectively, (P<0.05). CONCLUSION: Hepatic SEC injury occurs earlier than hepatic parenchymal cells during AOC. Recruitments of circulating neutrophils in the hepatic sinusoidal space might mediate the SEC injury, and ICAM-1 in the liver may modulate the PMN of accumulation. PMID- 11925622 TI - Nuclear factor kB activity in patients with acute severe cholangitis. AB - AIM: To determine the NF-kB activity in peripheral blood mononuclear cells (PBMC) in patients with acute cholangitis of severe type (ACST) and correlate the degree of NF-kB activation with severity of biliary tract infection and clinical outcome. METHODS: Twenty patients with ACST were divided into survivor group (13 cases) and nonsurvivor group (7 cases). Other ten patients undergoing elective gastrectomy or inguinal hernia repair were selected as control group. Peripheral blood samples were taken 24 hours postoperatively. PBMC were separated by density gradient centrifugation, then nuclear proteins were isolated from PBMC, and Electrophoretic Mobility Shift Assay (EMSA) used determined. The results were quantified by scanning densitometer of a Bio-Image Analysis System and expressed as relative optical density (ROD). The levels of TNF-alpha, IL-6, and IL-10 in the plasma of patients with ACST and healthy control subjects were determined by using an enzyme-linked immunoassay (ELISA). RESULTS: The NF-kB activity was 5.02 +/- 1.03 in nonsurvivor group, 2.98 +/- 0.51 in survivor group and 1.06 +/- 0.34 in control group. There were statistical differences in three groups (P<0.05). The levels of TNF-alpha and IL-6 in plasma were (498 +/- 53)ng.L(-1)and (587 +/- 64)ng.L(-1)in nonsurvivor group, (284 +/- 32)ng.L(-1) and (318 +/- 49)ng.L(-1)in survivor group and (89 +/- 11)ng.L(-1) and (102 +/-13)ng.L(-1)in control group. All patients with ACST had increased levels of TNF-alpha and IL-6, which were many-fold greater than those of control group, and there was an evidence of significantly higher levels in those of nonsurvivor group than that in survivor group (P<0.05). The levels of IL-10 in plasma were (378+/-32)ng.L(-1), (384+/ 37)ng.L(-1) and (68+/-11)ng.L(-1) in three groups, respectively. All patients had also increased levels of IL-10 when compared with control group (P<0.05), but the IL-10 levels were not significantly higher in nonsurvivors than in survivors (P>0.05). CONCLUSION: NF-kB activity in PBMC in patients with ACST increases markedly and the degree of NF-kB activation is correlated with severity of biliary tract infection and clinical outcome. PMID- 11925623 TI - Effect of areca on contraction of colonic muscle strips in rats. AB - AIM: To investigate the effects of areca on the contractile activity of isolated colonic muscle strips in rats and mechanism involved. METHODS: Each strip (LMPC, longitudinal muscle of proximal colon; CMPC, circular muscle of proximal colon; LMDC, longitudinal muscle of distal colon; CMDC, circular muscle of distal colon.) was suspended in a tissue chamber containing 5 mL Krebs solution (37 degrees C), bubbled continuously with 950 mL.L(-1) O(2) and 50 mL.L(-1) CO(2). The mean contractile amplitude (A), the resting tension (T), and the contractile frequency (F) were simultaneously recorded on recorders. RESULTS: Areca dose dependently increased the mean contractile amplitude, the resting tension of proximal and distal colonic smooth muscle strips in rats (P<0.05). It also partly increased the contractile frequency of colonic smooth muscle strips in rats (P<0.05). The effects were partly inhibited by atropine (the resting tension of LMPC decreased from 0.44 +/- 0.12 to 0.17 +/- 0.03; the resting tension of LMDC decreased from 0.71 +/- 0.14 to 0.03 +/- 0.01; the mean contractile amplitude of LMPC increased from -45.8 +/- 7.2 to -30.5 +/- 2.9; the motility index of CMDC decreased from 86.6 +/- 17.3 to 32.8 +/- 9.3; P<0.05 vs areca), but the effects were not inhibited by hexamethonium (P>0.05). CONCLUSION: Areca stimulated the motility of isolated colonic smooth muscle strips in rats. The stimulation of areca might be relevant with M receptor partly. PMID- 11925624 TI - Identification and characterization of a novel isoform of hepatopoietin. AB - AIM: To isolate a novel isoform of human HPO (HPO-205) from human fetal liver Marathon-ready cDNA and characterize its primary biological function. METHODS: 5' RACE (rapid amplification of cDNA 5' ends) was used to isolate a novel isoform of hHPO in this paper. The constructed pcDNA(HPO-205), pcDNA(HPO) and pcDNA eukaryotic expression vectors were respectively transfected by lipofectamine method and the stimulation of DNA synthesis was observed by (3)H-TdR incorporation assay. Proteins extracted from different cells were analyzed by Western blot. RESULTS: A novel isoform of hHPO (HPO-205) encoding a 205 amino acid ORF corresponding to a translated production of 23 kDa was isolated and distinguished from the previous HPO that lacked the N-terminal 80 amino acids. The dose-dependent stimulation of DNA synthesis of HepG2 hepatoma cells by HPO 205 demonstrated its similar biological activity with HPO in vitro. The level of MAPK (Mitogen-activated protein kinase) phosphorylation by Western blot analysis revealed that HPO-205 might have the stronger activity of stimulating hepatic cell proliferation than that of HPO. CONCLUSION: A novel isoform of hHPO (HPO 205) was isolated from hepatic-derived cells. The comparison of HPO-205 and HPO will lead to a new insight into the structure and function of hHPO, and provide the new way of thinking to deeply elucidate the biological roles of HPO/ALR. PMID- 11925626 TI - Protective effect of estradiol on hepatocytic oxidative damage. AB - AIM: To examine the protective effect of estradiol on the cultured hepatocytes under oxidative stress. METHODS: Hepatocytes of rat were isolated by using perfusion method, and oxidative stress was induced by a serum-free medium and FeNTA. MDA level was determined with TBA method. Cell damage was assessed by LDH assay. Apoptosis of hepatocytes was assessed with cytoflowmetric analysis. Expression of Bcl-xl in cultured hepatocytes was detected by Western blot. The radical-scavenging activity of estradiol was valued by its ability to scavenge the stable free radical of DDPH. RESULTS: Oxidative stress increased LDH from 168 +/- 25 x 10(-6)IU.cell(-1) to 780 +/- 62 x 10(-6)IU.cell(-1) and MDA(from 0.28 +/ 0.07 x 10(-6)nmol.cell(-1) to 1.35 +/- 0.12 x 10(-6)nmol.cell(-1)) levels in cultured hepatocyte, and estradiol inhibited both LDH and MDA production in a dose dependent manner. In the presence of estradiol 10(-6)mol.L(-1), 10( -7 )mol.L(-1) and 10(-8)mol.L(-1),the LDH levels are 410 +/- 53 x 10(-6)IU.cell(-1) (P<0.01 vs oxidative group), 530 +/- 37 X 10(-6)IU.cell(-1 ) (P<0.01 vs oxidative group), 687+/-42 x 10(-6)IU.cell(-1) (P<0.05 vs oxidative group) respectively, and the MDA level are 0.71+/-0.12 x 10(-6)nmol.cell(-1) (P<0.01 vs oxidative group),0.97+/-0.11 x 10(-6)nmol.cell(-1 )(P<0.01 vs oxidative group) and 1.27+/ 0.19 x 10(-6)nmol.cell(-1) respectively. Estradiol suppressed apoptosis of hepatocytes induced by oxidative stress, administration of estradiol(10( 6)mol/L)decreased the apoptotic rate of hepatocytes under oxidative stress from 18.6 +/- 1.2% to 6.5 +/-2.5%, P<0.01. Bcl-xl expression was related to the degree of liver cell damage due to oxidative stress, and estradiol showed a protective action. CONCLUSION: Estradiol protects hepatocytes from oxidative damage by means of its antioxidant activity. PMID- 11925625 TI - Telomere and telomerase in the initial stage of immortalization of esophageal epithelial cell. AB - AIM: To search for the biomarker of cellular immortalization, the telomere length, telomerase activity and its subunits in cultured epithelial cells of human fetal esophagus in the process of immortalization. METHODS: The transgenic cell line of human fetal esophageal epithelium (SHEE) was established with E(6)E(7) genes of human papillomavirus (HPV) type 18 in our laboratory. Morphological phenotype of cultured SHEE cells from the 6th to 30th passages, was examined by phase contrast microscopy, the telomere length was assayed by Southern blot method, and the activity of telomerase was analyzed by telomeric repeat amplification protocol (TRAP). Expressions of subunits of telomerase, hTR and hTERT, were assessed by RT-PCR. DNA content in cell cycle was detected by flow cytometry. The cell apoptosis was examined by electron microscopy (EM) and TUNEL label. RESULTS: SHEE cells from the 6th to 10th passages showed cellular proliferation with a good differentiation. From the 12th to the 16th passages, many senescent and apoptotic cells appeared, and the telomere length sharply shortened from 23kb to 17kb without expression of hTERT and telomerase activity. At the 20th passage, SHEE cells overcame the senescence and apoptosis and restored their proliferative activity with expression of telomerase and hTERT at low levels, but the telomere length shortened continuously to the lowest of 3kb. After the 30th passage cells proliferation was restored by increment of cells at S and G2M phase in the cell cycle and telomerase activity expressed at high levels and with maintenance of telomere length. CONCLUSION: At the early stage of SHEE cells, telomeres are shortened without expression of telomerase and hTERT causing cellular senescence and cell death. From the 20th to the 30th passages, the activation of telomerase and maintenance of telomere length show a progressive process for immortalization of esophageal epithelial cells. The expression of telomerase may constitute a biomarker for detection of immortalization of cells. PMID- 11925627 TI - Establishment of an artificial beta-cell line expressing insulin under the control of doxycycline. AB - AIM: Artificial beta-cell lines may offer an abundant source of cells for the treatment of type I diabetes, but insulin secretion in beta-cells is tightly regulated in physiological conditions. The Tet-On system is a "gene switch" system, which can induce gene expression by administration of tetracycline (Tet) derivatives such as doxcycline (Dox). Using this system, we established 293 cells to an artificial cell line secreting insulin in response to stimulation by Dox. METHODS: The mutated proinsulin cDNA was obtained from plasmid pcDNA3.1/C-mINS by the polymerase chain reaction (PCR), and was inserted downstream from the promoter on the expression vector pTRE2, to construct a recombined expression vector pTRE2mINS. The promoter on pTRE2 consists of the tetracycline-response element and the CMV minimal promoter and is thus activated by the reverse tetracycline-controlled transactivator (rtTA) when Dox is administrated. pTRE2mINS and plasmid pTK-Hyg encoding hygromycin were co-transfected in the tet293 cells, which express rtTA stably. Following hygromycin screening, the survived cells expressing insulin were selected and enriched. Dox was used to control the expression of insulin in these cells. At the levels of mRNA and protein, the regulating effect of Dox in culture medium on the expression of proinsulin gene was estimated respectively with Northern blot, RT-PCR, and radioimmunoassay. RESULTS: From the 28 hygromycin-resistant cell strains, we selected one cell strain (tet293/Ins6) secreting insulin not only automatically, but in response to stimulation by Dox. The amount on insulin secretion was dependent on the Dox dose (0,10,100,200,400,800 and 1000 microg.L(-1)), the level of insulin secreted by the cells treated with Dox (1000 microg.L(-1)) was 241.0pU.d(-1).cell(-1) , which was 25-fold that of 9.7pU.d(-1).cell(-1) without Dox treatment. Northern blot analyses and RT-PCR further confirmed that the transcription of insulin gene had already been up-regulated after exposing tet293/Ins6 cells to Dox for 15 minutes, and was also induced in a dose-dependent manner. However, the concentration of insulin in the media did not increase significantly until 5 hours following the addition of Dox. CONCLUSION: Human proinsulin gene was transfected successfully and expressed efficiently in 293 cells, and the expression was modulated by tetracycline and its derivatives, improving the accuracy, safety, and reliability of gene therapy, suggesting that conditional establishment of artificial beta-cells may be a useful approach to develop cellular therapy for diabetes mellitus. PMID- 11925628 TI - Epidemiological survey of cryptosporidiosis in Anhui Province China. AB - AIM: To provide scientific evidence for prevention and controlling of cryptosporidiosis, the infection of Cryptosporidium parvum and its epidemiological characteristics were studied in some areas of Anhui Province. METHODS: The oocyst of Cryptosporidium parvum in 5421 fresh stool samples from eleven areas of Anhui Province was tested by auramine-phenol stain and improved anti-acid stain respectively. The specific antibody of IgG, IgM and T subsets of 41 patients with positive Cryptosporidium parvum in stools were detected by ELISA and biotin-streptavidin (BSA) respectively. RESULTS: The total infective rate of Cryptosporidium parvum was 1.33% (74/5421). Among them, the positive rates of oocyst in the areas of Huaibei (1.82%) and Fuyang (1.80%) were higher. The positive rates of oocyst in stools of infants, pupils, middle school students, college students, adults, patients with diarrhea, and those with immunodeficiency were 3.15%(28/889), 0.82% (9/1098), 0.82%(9/1092), 0.83%(8/969), 0.85% (9/1095), 2.88%(8/278) and 8.33%(3/36)% respectively. The positive rates of oocyst in infants and the patients with diarrhea and immunodeficiency were significantly higher than those in controls (P<0.01). The positive rate of oocyst in males was similar to that in females (P>0.05). The positive rate of oocyst in urban areas (1.13%) was significantly lower than those in rural areas (1.72%, P<0.01). The positive rates of specific IgG, IgM and IgG+IgM in sera of the patients with positive oocyst in stool were 63.4% (26/41), 17.1% (7/41), 19.5% (8/41) respectively. The number fractions of T subsets of CD(3)(+), CD(4)(+), CD(8)(+) and CD(4)(+)/CD(8)(+) of the patients were 0.66+/-0.07, 0.44+/-0.06, 0.28+/-0.04 and 1.58+/-0.32 respectively. The difference between the patients and the controls was significant (P<0.05). The main manifestations of the patients were subclinical infection, in forms of slight abdominal pain, mild diarrhea, and loose stool. CONCLUSION: There are two infection peaks in infection of Cryptosporidium parvum and its infection can be found more often in infants, patients with diarrhea or immunodeficiency, and in rural areas. Subclinical infection is the main manifestation and might be easily misdiagnosed. When the therapeutic effectiveness is low for diarrhea, the infection of Cryptosporidium parvum should be considered, concerning their age and immune function. PMID- 11925630 TI - Quantitative analysis of transforming growth factor beta 1 mRNA in patients with alcoholic liver disease. AB - AIM: To investigate the expression of the transforming growth factor beta 1(TGF beta 1) mRNA in different stages of alcoholic liver disease (ALD) and its clinical value. METHODS: One hundred and seven male alcoholics were grouped by clinical findings into four groups: alcohol abusers without liver impairment (n =22), alcoholic steatosis (n =30); alcoholic hepatitis (n=31); and alcoholic cirrhosis(n=24). Using peripheral blood mononuclear cells (PBMC) as samples the gene expression of TGF-beta 1 was examined quantitatively by reverse transcription polymerase chain reaction (RT-PCR) and dot blot. There are 34 healthy subjects served as control. RESULTS: The expression of TGF-beta 1 from all ALD patients was significantly greater than that in controls (1.320 +/- 1.162 vs 0.808 +/- 0.276, P<0.001). The differences of the expressions were significant between the patients from each groups (alcoholic steatosis, alcoholic hepatitis and alcoholic cirrhosis) and the controls (1.168 +/- 0.852, 1.462 +/- 1.657, 1.329 +/- 0.610 vs 0.808 +/- 0.276, P<0.050). No significant differences of TGF beta 1 mRNA expression were observed between alcohol abusers without liver impairment and controls. The expressions in patients with alcoholic hepatitis and alcoholic cirrhosis were significantly greater than that in alcohol abusers respectively (1.462 +/- 1.657, 1.329 +/- 0.610 vs 0.841 +/- 0.706, P<0.050). No significant differences of TGF-beta 1 mRNA expression were observed between alcoholic fatty liver men and alcohol abusers. CONCLUSION: TGF-beta 1 expression level can be a risk factor for alcoholic liver disease and might be related to the inflammatory activity and fibrosis of the liver in patients. PMID- 11925629 TI - Gastrin, somatostatin, G and D cells of gastric ulcer in rats. AB - AIM: To investigate the relationship among gastrin, somatostatin, G and D cells in gastric ulcer and in its healing process in rats. METHODS: Fourty-nine Wistar rats were divided into 7 groups. The gastric ulcer model was induced by acetic acid successfully. The gastrin and the somatostatin in rat plasma, gastric fluid and antral tissue were measured by radioimmunoassay(RIA). G and D cells in antral mucosa were analyzed with polyclonal antibody of gastrin and somatostatin by immunohistochemical method and Quantimet 500 image analysis system. RESULTS: In gastric ulcer, the level of gastrin in plasma, gastric fluid, and antral tissue increased, that of somatostatin declined, and the disorder gradually recovered to the normal level in the healing process. Immunohistochemical technique of G and D cells in antral mucosa demonstrated that the number of G cells increased and that of D cells decreased, both areas of G and D cells declined, the ratio of number and area of G/D increased in gastric ulcer, and the disorder gradually recovered in the healing process. CONCLUSION: In gastric ulcer, the increased gastrin secreted by G cells, the declined somatostatin secreted by D cells, and the disordered G/D cell ratio can lead to gastrointestinal dysfunction. PMID- 11925631 TI - Assessment of duodenogastric reflux by combined continuous intragastric pH and bilirubin monitoring. AB - AIM: To assess the diagnostic value of a combination of continuous intragastric pH and bilirubin monitoring in the detection of duodenogastric reflux (DGR), and the effects of diet on the bilirubin absorbance. METHODS: 30 healthy volunteers were divided into two groups: standard diet group (Group 1) 18 cases, free diet group (Group 2)12 cases. Each subjects were subjected to simultaneous 24-hour intragastric pH and spectrophotometric bilirubin concentration monitoring (Bilitec 2000). RESULTS: There was no difference of preprandial phase bilirubin absorbance between two groups. The absorbance of postprandial phase was significantly increased in group 2 than group 1. There was no difference between preprandial phase and postprandial phase absorbance in group 1. Postprandial phase absorbance was significantly higher in group 2. In a comparison of bile reflux with intragastric pH during night time, there were 4 types of reflux: Simultaneous increase in absorbance and pH in only 19.6%, increase in bilirubin with unchanged pH 33.3%, pH increase with unchanged absorbance 36.3%, and both unchanged in 10.8%. Linear regression analysis showed no correlation between percentage total time of pH<4 and percentage total time of absorbance>0.14, r=0.068 P<0.05. CONCLUSION: Because of the dietary effect, high absorbance fluids or foods should be avoided in detection. Intragastric pH and bilirubin monitoring separately predict the presence of duodenal (and/or pancreatic) reflux and bile reflux. They can not substitute for each other. The detection of DGR is improved if the two parameters are combined simultaneously. PMID- 11925634 TI - Create a family-supportive culture. PMID- 11925635 TI - Have a talk. PMID- 11925636 TI - A perfect union? Organized labor & EMS. PMID- 11925637 TI - The nuts and bolts of job hunting. PMID- 11925638 TI - Living & breathing case studies. PMID- 11925639 TI - Assessment of the altered mental status patient. AB - Patients presenting with an altered mental status are a fairly common event for the EMS provider. Much of the uncertainty in etiology and the ambiguity in defining the clinical presentation can be minimized with a good understanding of the common pathology associated with AMS and applying a well-organized, thorough approach to assessing these patients. Recognizing the differences between cognition and arousal and correlating the anatomical functions of the brain to the symptom presentation help considerably in understanding the causes of AMS. Classifying causes as arising from a structural nature or attributing the AMS to either a metabolic or a toxic source are fundamental in understanding potential etiologies. In assessing and reporting clinical findings, it is important to eliminate collective phrases to represent the patient's mental status and, instead, describe the patient's behavior and responses to stimuli. Complying with a standardized approach to assessment and employing effective assessment aids (such as AVPU or SAMPLE) help to prevent omissions of relevant data and contribute to a comprehensive assessment. With these goals in mind, the emergency care provider can effectively assess and manage patients with altered mental status with confidence and precision, and in a comprehensive manner. PMID- 11925640 TI - There are no ordinary drunks. The alcoholic patient. PMID- 11925641 TI - [Undiagnosed primary Boerhaave syndrome]. AB - Spontaneous rupture of the oesophagus (Boerhaave syndrome) is a rare acute condition. Successful treatment of this life threatening condition depends on early diagnosis, the patient's general condition and subsequent treatment. The authors describe two cases of successful treatment of patients with primarily unrecognized spontaneous perforation of the oesophagus. PMID- 11925642 TI - [Pathology of the spleen in children--current possibilities of surgical treatment]. AB - In the submitted paper the authors review the contemporary state of surgery of the spleen in child age. Surgical treatment is used most frequently in injuries of the spleen, in portal hypertension and in haematological diseases. In a group of patients during the last decade efforts to preserve the spleen are emphasized and experience is reported with the use of therapeutic procedures. The authors emphasize that splenectomy in children should be an exceptional operation. PMID- 11925643 TI - [Surgical treatment of children with necrotizing enterocolitis]. AB - Necrotizing enterocolitis is an acute neonatal disease. It affects in particular premature neonates with a birth weight lower than 1500 g. Despite extensive research the etiology of the disease remains obscure. The majority of authors assume multifactorial causes. Research workers try to detect various laboratory and clinical factors which could serve as criteria for surgical intervention. The sensitivity and specificity of these tests and laboratory examinations detect the disease only in an advanced stage and in the stage of perforation of the gastrointestinal tract (GIT). The main surgical approach to the treatment of this disease remains laparotomy and resection of the necrotic portion of the gut with enterostomy. In a limited number of children resection of a portion of the necrotic gut with primary anastomosis is possible. In some very serious conditions and in infants with a birth weight below 1000 g peritoneal drainage and subsequent "second look" surgery should be sufficient. It is a disease with calls for a maximum individual approach and there are no accurate defined instructions for surgical treatment. The surgeon's experience and the standard of preoperative and postoperative care are decisive. PMID- 11925644 TI - [Surgical treatment of discogenic compression of the spinal cord in the thoracic region]. AB - The objective of the submitted work was to draw attention to some problems associated with the diagnosis and treatment of prolapse of sequestra of thoracic intervertebral discs. The investigated group comprised 9 subjects (4 women, 5 men) aged 33 to 67 years operated upon at the Neurosurgical Clinic in Kosice between Jan. 1 1982 and June 30 2001 on account of compression of nervous structures in the thoracic portion of the spine by sequestra of intervertebral discs. This was manifested by back pain, a sensation of stiffening of the muscles of the lower extremities, altered sensitivity and in all by impaired gait. Only one female patient developed urinary retention, another one painless paraparesis of the lower extremities, therefore the condition was evaluated as a demyelinisation process. In three patients as the only imaging examination method perimyelography was used, in another two it was supplemented by CT. Four patients were examined by MRI. This graphic method proved to be the most suitable. In all subjects of the investigated group the clinical picture and examination methods indicated a unilateral predominance of the affection. In five subjects it proved possible to remove the sequestream of the intervertebral disc via laminectomy, in another four a transpedicular approach into the spinal canal was used successfully. In all patients the condition improved after surgery. PMID- 11925645 TI - [Analysis of complications in the treatment of fracture leg fractures with unreamed intramedullary nailing]. AB - Based on their group of 52 patients the authors analyze the complications they encountered in the treatment of crural fractures by a not predrilled intramedullary nail. Consistent with the literature they found that this method is successful in the treatment of closed and open fractures of the crural diaphysis. They mention, however, a relatively high percentage of complications which in the long run are not a impediment for healing of the fracture but influence it. In the discussion these complications are analyzed in detail in an attempt to find a solution and to avoid them in future patients. They conclude that even this relatively simple and effective method of fractures of the crural diaphysis has its pitfalls and its indication and implementation call for much attention. PMID- 11925646 TI - [Personal experience with laparoscopic cholecystectomy in acute cholecystitis]. AB - Authors refer about their experiences with operations of acute cholecystitis by laparoscopy. They have evaluated retrospectively 1600 cases of laparoscopic cholecystectomies (LCHE) realized in the period from 1994 to 2000. In 302 (18.8%) cases was realized laparoscopic cholecystectomy for acute cholecystitis, which was confirmed by histopathological finding. During the mentioned period a total number of laparoscopic cholecystectomies for acute cholecystitis (n = 302) rise a number of classical cholecystectomies for the same diagnosis (n = 227). Very important role by authors take early dispatching a patient to a department of surgery and early estimation the diagnosis with consecutive laparoscopic treatment. Rates of peroperation, postoperation complications and reoperations in laparoscopic surgery of acute cholecystitis is comparable with elective laparoscopic cholecystectomy. PMID- 11925647 TI - [Liver abscess--a rare complication of ventriculoperitoneal drainage--case report]. AB - The authors present liver abscess as the rare complication of a ventriculoperitoneal drainage. Diagnostics includes combination of USG and CT examination. The treatment requires the evacuation of an abscess cavity temporary followed by the externalization of a ventriculoperitoneal shunt and antibiotic therapy. This complication is important to think about in the differential diagnostics of acute fever, abdominal pain and general signs of sepsis in a patient with ventriculoperitoneal shunt. PMID- 11925648 TI - [Pelvic pain--appendiceal diseases and laparoscopic treatment]. AB - The authors describe the diagnostic problem of "pelvic pain" by the laparoscopic approach. They draw attention to the high percentage of appendicopathies revealed during laparoscopic revisions on account of this diagnosis. They describe their own laparoscopic procedure, its advantages ad universal application which makes it possible to perform not only laparoscopic appendectomies but also other laparoscopic operation of organs of the pelvis minor as well as cholecystectomies. In the discussion they draw attention to the important number of other findings which usually are not detected during classical operations and where the laparoscopic approach is also important. PMID- 11925649 TI - [Maffucci's syndrome--case report of a 5-year-old female patient]. AB - Maffucci's syndrome is a congenital non-hereditary syndrome characterized by a combination of dyschondroplasia (enchondromatosis) and haemangiomatosis. It is a rare disease. During the last 140 years only 200 cases were described [5]. 109 cases are described in the literature by other authors [21]. This disease is associated with a high risk of development of neoplastic processes, in particular sarcomatous transformation of enchondromatoses as well as other mesodermal and non-mesodermal malignities [13]. Based on the incidence of spinocellular haemangioendothelioma with other congenital syndromes, the presence at a young age of patients and the multicentric incidence support the assumption that spinocellular haemangioendothelioma may be the manifestation of genetically based mesodermal disease which may be associated with the picture of Maffucci's syndrome [6]. In their case-history the authors present a 5-year-old patient. They describe the clinical course and findings taking into account possible manifestations and risks associated with the disease. With regard to the low incidence of the disease and its interdisciplinary character the authors pen the question of possible causal, symptomatic or palliative treatment. PMID- 11925650 TI - [Developments in surgery of the extracranial circulation--findings from a vascular surgery department]. AB - The objective of the paper is to summarize hitherto assembled experience of the department with the application of new approaches to reconstruction operations of the extracranial circulation with emphasis on its most important part, i.e. the carotid circulation. The main feature of this approach is an effort to increase the safety of the operation for the patient at risk which implies among others extension of indications for reconstruction surgery. PMID- 11925651 TI - [Bronchopleural fistula after anatomic pulmonary resection]. AB - OBJECTIVE: The authors evaluate in a retrospective study the incidence of bronchopleural fistulae after lung resections implemented in their department in the course of 12 years, and reflect on the causes of their development. METHOD: They compare their experience with mechanical and manual suture of the large bronchi in relation to the incidence of fistulae, with experience assembled by other departments. RESULTS: While preferring mechanical suture of the bronchus, they achieved in a group of 1601 pulmonary resections a 0.87% incidence of bronchopleural fistulae whereby after mechanical suture a fistula was recorded in 0.74% and after manual suture in 2.52%. CONCLUSION: Based on the results of their observations which they compare with work of others, the authors assume that if the department has adequate experience, impaired healing of the bronchus is rare regardless whether the manual or mechanical technique of suture is used. PMID- 11925652 TI - [Traumatic rupture of the diaphragm--case report]. AB - The authors describe the case-history of a patient with traumatic rupture of the diaphragm where clinical manifestation occurred five years after the primary injury. PMID- 11925653 TI - [Invagination--the first manifestation of leiomyosarcoma]. AB - The authors describe in their paper one of the less frequent ileus acute abdominal events in adult age--invagination. At the same time they comment also on the rare cause of invagination, a leiomyosarcoma of the small intestine. In the discussion they mention the relationship of this tumour to gastrointestinal stromal tumours. PMID- 11925654 TI - [Laparoscopic transperitoneal adrenalectomy in hormonally-active adrenal tumors]. AB - Laparoscopic adrenalectomy is considered the "gold standard" in adrenal surgery for benign functioning (and non-functioning) tumors. Laparoscopy meets criteria of miniinvasive surgery with minimal postoperative pain, short hospital stay and an outstanding cosmetic effect, the radicality of the procedure, open surgery and laparoscopy are equally effective. In cooperation with endocrinological department focused on detection of functioning adrenal tumors, the diagnostic and therapeutic procedure seems to be very efficient, so that the primary goal of rapid cure could be achieved. Patients after laparoscopic adrenalectomy mostly do not need chronic antihypertensive medication and only those after bilateral adrenalectomy (Cushing's disease) require life-long low dose hydrocortisone therapy. In pheochromocytomas, total excision of the tumor is considered to be a life-saving procedure, although up to 20% may recur as hereditary or as a part of von Hippel-Lindau's disease. PMID- 11925655 TI - [The plug system and laparoscopic hernioplasty in recurrent inguinal hernia]. AB - A total of 1511 patients were operated on at the 1st Department of Surgery Charles University in Prague during 1996 through 2000. Of this number, 81.3% underwent surgery for primary and 18.7% for recurrent inguinal hernia. Among the patients with recurrences, 81% had the first, 15% patients the second, and 4% at least the third episode of recurrent hernia. A total of 604 patients were operated on during 1999-2000, when a plug system (Bard Mesh Perfix Plug) was introduced into surgery protocols. Of this number, 113 patients had a recurrent hernia with an identical ratio of recurrences. The following plastic surgery interventions were carried out during the latter period: McVay-Lotheissen (54.2%), TAPP (26.5%), PHS (13.2%), Plug (3.3%), and Lichtenstein (2.6%). The following interventions were used when operating recurrences: McVay-Lotheissen (20.3%), TAPP (31%), PHS (21.2%), Plug (16.8%), and Lichtenstein (9.7%). During the 1-24-month follow-up period, recurrences occurred 1x after the TAPP procedure, 1x after McVay-Lotheissen, and 1x after the Lichtenstein procedure (95% and 88% patients who underwent plug and Lichtenstein procedures, respectively, were included in the follow-up). Comparison of plug vs. TAPP in patients with recurrent hernia (Plug/TAPP): mean age of patients: 62/45 years, length of operation: 66/48 minutes, overall post-operative morbidity: 9.4%/3.6%, hospitalization: 4.3/1.8 days, return to the working process 28 days (range 9-38 days) vs. 7.2 days (range 2-15 days). Both procedures can be considered safe and reliable interventions for treatment of recurrent inguinal hernia. They meet the requirements for elastic strength (more the plug procedure), closure, and bridging of defects even in several layers. The plug system can be implanted under local anaesthesia, it is capable of bridging large defects in a firm and elastic manner, and appears to be a very suitable solution for large defects in patients with advanced biological age. In these indications, the plug system brings many benefits but also the risk of more open access and greater "quantity" of materials. TAPP appears to be more suitable in younger patients with recurrences and large defects. We evaluate favourably in particular, all aspects of the postoperative period. An experienced team of surgeons is needed to achieve good results in both procedures. PMID- 11925656 TI - [A pediatrician's views on the treatment of extensive hemangiomas in childhood]. AB - Haemangiomas are the most frequent tumours of child age. They are found in 10-12% of infants. The authors summarize contemporary knowledge of the treatment of haemangiomas as well as their own experience with their treatment. They follow-up 18 children with haemangiomas at different sites, incl. ten with extensive haemangiomas. The latter were treated conservatively with prednisone and interferon alpha. PMID- 11925657 TI - [Use of ultrasonography in surgery]. AB - OBJECTIVE: To draw attention to the perspectives of application of ultrasonography in surgery. METHOD: Based on their own experience and data in the literature the authors draw attention to contemporary possibilities to use ultrasound in surgery. This concerns in particular the following areas: abdominal sonography, injuries, examination of small areas, peroperative sonography, laparoscopic sonography, intraluminal sonography, Dopplerometric examination of the vessels, use during transplantations and in neurosurgery. RESULTS AND CONCLUSIONS: While urologists use sonographic examinations commonly, only a small percentage of surgeons are familiar with this method. The authors explain the necessity of greater involvement of surgeons in this area which is consistent with the development of surgery and trends abroad. PMID- 11925658 TI - [Initial clinical experience with Omniflow II, a bio-textile prosthesis]. AB - The authors give an account of their first clinical experience with vascular prostheses Omniflow II which are available only recently and should be used for vascular reconstructions for arterial obliterations at the site of peripheral aortal affections. PMID- 11925659 TI - [Infectious aneurysm of the abdominal aorta caused by Salmonella: diagnosis, endovascular therapy]. AB - The authors present a case of an infected abdominal aortic aneurysms by means of a minimally invasive endovascular method using a stent graft. This patient had already been treated by TIPS (transjugular intrahepatic portosystemic shunt) for repeated varicose bleeding due to liver cirrhosis. Standard surgical therapy of infected abdominal aneurysms and about advantages and possibilities of endovascular treatment with stent grafts are discussed. Regular follow-up of the patient and long-term antibiotics therapy are important for accurate assessment of an implanted stent graft into the infected abdominal aorta. Endovascular stent graft combined with antibiotic therapy may be an alternative to conventional open surgery in managing infected infrarenal abdominal aneurysms. PMID- 11925661 TI - [Venous thrombosis and pulmonary embolisms in air travel]. AB - The risk of venal thrombosis and pulmonary embolia during aviation is rather small. The risk markedly rises during long flights (over 7500 km) and namely in elderly persons. Danger of the disease can be lowered by lower limbs exercising, liberal intake of soft drinks, and by elastic calf bandages. Persons of the high risk are candidates for low-molecular heparin administration. PMID- 11925660 TI - [Cervical lymphangiectasia--case report]. AB - A case-report of a patient with paraesthesia of the left upper extremity caused by a tumor localized in the left supraclavicular area is presented. The patient underwent an operation. Two formations--a lipoma and cyst--were found. Both were removed and histologically examined. The diagnosis of lipoma and neck lymphangiectasia was made. On the third postoperative day the patient was discharged home with no more paraesthetic complaints. A check-up after three months did not reveal any complaints present before operation. In the paper the differential diagnosis of neck tumors with a view to resistance associated with the lymphatic system especially lymphangioma, lymphangiectasia and hygroma coli cysticum is discussed. Lymphangioma is a benign oncogenic affection of lymphatic vessels developing during the postnatal period of life. Lymphangiectasia is a congenital disease without proliferating activity. Hygroma coli cysticum is a congenital extensive multioccular formation localized in the neck area. All three affections are often incorrectly mistaken for each other. Histological examination is decisive for the correct diagnosis. PMID- 11925662 TI - [Gastrointestinal tract dysfunction in critical illness]. AB - Until relatively recently, the gastrointestinal (GI) tract was considered a dormant, metabolically and immunologically inactive organ in critically illnesses. However, the GI tract provides a number of crucial functions that, in fact, may influence morbidity and mortality of many critically ill patients. Its large absorptive area provides a site for nutrient digestion and utilization and serves as an important barrier preventing the systemic absorption of intraluminal microbes and its toxic products. Moreover, the GI tract is the largest reservoir of lymphocytes in the body, which significantly contribute to the immune response of the critically ill patients. The gut dysfunction occurs frequently and early in the intensive care patients. Abnormal colonization, impaired intestinal epithelial barrier function and bacterial translocation represent the key components of gut failure implicating in the pathogenesis of sepsis and multiorgan dysfunction. This review summarizes recent insights into the role of the gut in critically ill patients with particular focus on 1) the basis of "gut origin hypothesis", 2) pathophysiology of gut dysfunction, 3) monitoring of intestinal function, and 4) protective measures and novel therapeutic strategies. PMID- 11925663 TI - [Disorders of mitochondrial energy metabolism in patients with the Kearns-Sayre syndrome]. AB - BACKGROUND: Kearns-Sayre syndrome is a multisystem disorder caused by rearrangements of mitochondrial genome including various deletions and/or duplications. The aim of the study is to analyse the impact of mitochondrial DNA (mtDNA) deletions on the mitochondrial energetic metabolism in five patients with Kearns-Sayre syndrome. METHODS AND RESULTS: The course of the disease is progressive in all patients. All of them have bilateral ptosis and external opthalmoplegia, four have retinitis pigmentosa, three have progressive muscle weakness and three have pacemaker because of complete A-V heart block. One patient underwent renal transplantation at the age of 12 because of a chronic renal failure. Southern blot analysis in muscle tissue revealed large scale heteroplasmic mtDNA deletions (3-7.4 kb) in all patients, the number of mutated copies of mtDNA ranged from 50 to 70%. Spectrophotometric measurements of respiratory chain complexes activities in muscle tissue revealed various combinations of defects of complex III, IV and I + III activities in all patients. Nevertheless, the lactic acidosis was permanently present only in one patient. Ragged-red fibers were found in two patients. CONCLUSIONS: Although the diagnostic of Kearns-Sayre syndrome is based on clinical features, molecular analysis of mtDNA is necessary to confirm the diagnosis. The prognosis of the disease is unfavourable and co-operation between the patient and various specialists is necessary for the treatment, which is currently only symptomatic. PMID- 11925664 TI - [Fatty acid composition of serum phospholipids and the effect of insulin in patients with type 2 diabetes]. AB - BACKGROUND: A direct relation has been found between the insulin sensitivity and fatty acid (FA) composition of serum and tissue lipids in adult humans. The aim of our study was to confirm the above relation in different groups of type 2 diabetic patients (DM2). METHODS AND RESULTS: The FA composition of serum phospholipids was measured by gas liquid chromatography at the insulin action during the two-step hyperinsulinemic isoglycemic clamp (1 and 10 mU/kg.min) in a group of 21 newly diagnosed DM2 subjects (DMN) and in 24 healthy controls (HC). The DMN group consisted of a subgroup of patients treated with hypoglycemic agents (DMH; n = 21) and a group treated with the diet only (DMD; n = 11). In diabetics, the metabolic clearance rates of glucose at either insulin level (MCRglusubmax and MCRglumax) were significantly reduced in comparison with HS (MCRglusubmax--DMN: 5.35 +/- 2.7 ml.kg-1.min-1; DMH: 5.38 +/- 2.17 ml.kg-1.min-1; DMD: 5.48 +/- 2.35 ml.kg-1.min-1; HS: 10.9 +/- 3.3 ml.kg-1.min-1; p < 0.01; and MCRglumax--DMN: 13.3 +/- 3.3 ml.kg-1.min-1; DMH: 12.5 +/- 3.0 ml.kg-1.min-1; DMD: 13.3 +/- 3.0 ml.kg-1.min-1; HS: 17.4 +/- 3.8 ml.kg-1.min-1; p < 0.05). An increased content of highly unsaturated n-6 family FA (p < 0.01), especially of the arachidonic acid (DMN: 10.98 +/- 1.79%; DMD: 10.78 +/- 1.64%; DMH: 10.97 +/- 1.70% vs HS: 8.51 +/- 1.53% (p < 0.01) was found in all groups of diabetics compared to HS, and lower levels of linoleic acid in DMN (p < 0.001), and DMH (p < 0.05) were identified. The content of saturated FA and that of monosaturated FA was comparable in HS, DMN and DMD persons. While in healthy subjects a negative correlation between MCRglu and the content of saturated FA as well as a positive association between insulin action and the ration of linoleic and arachinonic acids were found, no such relation was found in the diabetic subjects. CONCLUSIONS: Different groups type 2 diabetic patients show altered FA pattern of serum phospholipids. Changes in FA composition may play a role in the modulation of insulin action in peripheral tissues, but they cannot explain the insulin resistance in type 2 diabetic patients. PMID- 11925665 TI - [The present status of andrology in the Czech Republic]. AB - Andrology is defined as a branch of science and medicine dealing with the male reproductive organs in animals and in men and with the diseases of these organs. Included in this definition are all related specialized branches of science (Definition of the International Society of Andrology). This definition is very wide for the perspectives of andrology it is not optimal. In Czech Republic the Andrological Section of the Czech Urological Society was founded in March 1998. This section aims to disseminate knowledge on andrology by coordination and organization of congresses, conferences and courses in andrology, to encourage the development of basic and clinical research in andrology, to facilitate the collaboration by distribution information and to encourage sponsoring of publication related to the field of andrology. PMID- 11925666 TI - [A brief account of the 130 year history of the First Czech Internal Medicine Clinic]. AB - The first Czech medical clinic in Prague was founded in 1871 during the period of Czech revival by prof. Bohumil Eiselt. Since that time the clinic was headed by eight professors, each of them being an impressive and outstanding personality contributed significantly to its future development. The clinic keeps its immense vitality and represents the place of an outstanding curative and research activity. PMID- 11925667 TI - [What is and what is not informed consent]. AB - Coequalitisation of the relation between the physician and his patient, which respects the professional dominancy of the first one, has promoted the full guidance before any serious diagnostic or therapeutic intervention. Patient has the right to be informed of his health status and of the therapeutic plan. He has also the right to refuse such information. Only after the full information, the patients can express his approval with the plan for therapy. Before any intervention, the patient's approval is necessary; exceptional are only by the law implicated situations. To get the lawful and therefore the qualified or informed approval, the patient should be able to accept the information, to consider its significance, to make decision, and the express his will. Incapable of doing that is not only the person with permanently decreased ability to resolve, but also those who are not able to accept the information because of language problem and those, whose psychical state is influenced by the actual state of the disease, therapy etc. For a child, the person who has the right to be informed and to give the consent is his protector (usually his parent), however, within the limits of his comprehension an older child has also the right to express his opinion. For the common treatment the written form of approval is not necessary, it is enough to get the oral consent. For intervention of a higher significance and risks, the written form is recommended. It should be connected with a possibility to discuss and get explained any questions. The information should include not only what is to be done, but also how it will be done and what are the benefits of the treatment. It is essential to inform the patient also about the after-effects, side effects and possible complications, all that considering their significance and incidence. Information about the intervention and its risks does not limit the patient's right for compensatory damages, but it frequently improves communication and mutual collaboration between the patient and health care workers. PMID- 11925668 TI - [Breast cancer: an overview of factors affecting the onset and development of the disease]. AB - Breast cancer is the most common malignancy in women. The number of affected individuals is raising and the age at the onset has shifted towards younger age groups. Therefore, it is of great importance to improve primary and secondary prevention of breast cancer. New techniques of molecular biology significantly increased our understanding about the onset and progression of especially familial breast cancer. Further research in areas of molecular genetics and epidemiology is needed to find reliable diagnostic tools for screening of population groups to identify individuals at high risk of breast cancer. The follow up of interaction of various predisposing genes and environmental factors in breast cancer pathogenesis is of considerable public health importance as well. Some of risk factors covering areas of lifestyle and genetics are listed in this review as well as a survey of screening approaches and future prospects. PMID- 11925669 TI - [Depression and painful states and their treatment in oncology patients]. AB - Current trends in the treatment of oncologically diseased patients are aimed at comprehensive conception and at evaluation of all aspects influencing the health state of the patient including the psychical factors. Behavioral research studies the biological, psychological and social markers. It has brought more effective and more comprehensive therapeutical activities, including psychotherapy, sociotherapy and psychopharmacotherapy. It is sure that malignant tumors, their development and treatment represent a serious psychical stress. It is often connected with pain, severe body complains, and also with lowering or loss of the social position, reduction of social contacts and in serious cases with the threat of death. It can result an increasing dependence, anxiety, phobia and hopelessness. According to experts, the emotional reactions of oncologically diseased are similar to reactions of patients suffering from other chronic diseases. PMID- 11925670 TI - [Role of the steroids, SHBG, IGF-1, IGF BP-3 and growth hormone in glucose metabolism disorders during long-term treatment with low doses of glucocorticoids]. AB - BACKGROUND: The relationships between selected steroids, SHBG, growth hormone, IGF-1, IGF BP-3 and indicators of glucose metabolism were studied in the group of 20 female patients (15-20 yrs) on long-term treatment with low prednisone doses (< 0.3 mg/kg/day) (baseline phase) and after adding 1000 mg of metformin per day for following 6 months to improve impaired glucose metabolism (control phase). METHODS AND RESULTS: Lower basal DHEAS and DHEA (DHEA/S) levels were found as compared with reference values. Only DHEAS level returned into the reference range after the treatment with metformin. Decrease of DHEA/S depended on the doses (DHEAS -0.7621, DHEA -0.7685). Positive correlations between DHEA/S and of the results insulin tolerance were found as at the baseline (+0.4452, resp. +0.4455) as well as in the control period after the metformin administration (+0.7549, resp. +0.6073). Testosterone (T) and dihydrotestosterone(DHT) values were within the reference range during the whole study. Due to very low SHBG levels higher free androgen index (FAI) was recorded in more than half of the patients. Significant relationships were revealed between former gonadal androgens and indicators of glucose metabolism deterioration at the control phase: T correlated: with fasting insulin (+0.6005), with HOMAIR (+0.5380), with insulin/glucose (+0.5261), with fasting glucose (+0.9268), with AUC glucose (+0.6792), FAI: with fasting insulin (+0.5560), with HOMAIR (+0.5269), with fasting glucose (+0.9025), with AUC glucose (+0.7143), DHT: with fasting C peptide (+0.7921), with AUC C peptide (+0.7143). SHBG correlated: with fasting glucose (-0.6519), and with AUC glucose (-0.5868). The tendency of GH to lower, and IGF-1, IGF BP-3 to higher values at the baseline changed at the control phase: fasting and AUC value of GH increased (signif.), while were IGF-1 (nonsignif.) and IGF BP-3 (signif.) levels decreased. Surprisingly, no correlation was observed between GH and parameters of glucose metabolism. Contrary to GH, baseline IGFBP-3 values correlated: with HOMAIR (+0.5002), with insulin/glucose (+0.4860). The same relationships were found between AUC IGF BP-3 (+0.5676, +0.5559), IGF-1 (HOMAIR only +0.5412), IGF-1/IGF BP-3 (+0.5059, +0.5716) and parameters of insulin sensitivity (HOMAIR, insulin/glucose) in the control period. For the first time negative correlations between IGF-1, IGF-1 AUC, IGF BP-3, IGF-1/IGF BP-3 and somatostatin blood levels were discovered at the control phase. CONCLUSIONS: The study brought a number of new information about the importance of the "non-classical" glucoregulatory hormones in impairment of glucose metabolism, during long-term administration of low prednisone doses. The results suggest, that without normalisation of low DHEA/S, SHBG and high FAI levels it would not be possible to correct glucose metabolism properly in patients with long-term glucocorticoid therapy. PMID- 11925672 TI - Secured investments. How safe are your supplies and equipment? PMID- 11925671 TI - [Syphilis in the third millenium]. AB - BACKGROUND: For several decades the clinical picture of all stages of syphilis has been assumed to be immutable. The aim of the study was to describe the real spectrum of cutaneous and mucosal manifestations and organ changes and to ascertain whether the incidence of the disease has an increasing trend. METHODS AND RESULTS: The studied group included 92 patients (44 males, 48 females, average was 31.3 years) diagnosed in years 1992 to 1998 at our clinics. The diagnose was confirmed by a microscopic and/or serologic examination. Number of patients with the primary syphilis increased more than twice (1992 versus 1997), incidence of newly registered manifestations of the secondary syphilis has also increased (1996 to 1998), number of hospitalised patients in the stadium of syphilis latens increased at least twice (1992 versus 1996). Higher was the number of proven cases in pregnant patients: number of dead births increased twice, inborn forms of syphilis were three times more frequent. Among the newly described clinical forms the specific gastritis and specific uveitis were diagnosed. In immunosuppressed patients forms of lues maligna and the blue toes syndrome were identified. However, those forms were not diagnosed among our group of studied patients. CONCLUSIONS: During the last 10 years a significant increase of incidence of syphilis was reported in the population of Czech Republic. Such dangerous and almost explosive rise of new cases has been accompanied with increased number of inborn forms and with the occurrence of new clinical manifestations, namely in immunosuppressed patients. PMID- 11925673 TI - Process of elimination. Know what to ask when choosing a third-party reprocessor. PMID- 11925674 TI - Measuring Leapfrog's impact. PMID- 11925675 TI - Health system clicks with new MMIS. PMID- 11925676 TI - Stats & facts. The effect of today's practice environment on physician income. PMID- 11925678 TI - Internet technology and bioterrorism. PMID- 11925679 TI - Making the most appropriate formulary choices: a prescription for quality. PMID- 11925680 TI - Pharmacy benefit management as a quality safeguard. PMID- 11925681 TI - What is the most important component of disease management programs? PMID- 11925682 TI - The effect of donepezil therapy on health costs in a Medicare managed care plan. AB - The objective of this study was to estimate the ramifications of donepezil use on the health care costs of a large Medicare managed care plan. Patients with a diagnosis of Alzheimer's disease or related dementia were identified from the claims-and-encounter records of the plan. Costs for 204 patients identified as having Alzheimer's disease and who were receiving donepezil were compared with a control group of 204 patients with Alzheimer's disease who had matching characteristics, but who were not receiving therapy. After controlling for age, gender, pharmacy benefits, comorbid conditions, and complications of dementia, annual costs for medical services and prescription drugs were found to be $3,891 lower for the study group. Costs were $4,192 lower for patients receiving longer term therapy (> or = 270 day supply of donepezil) and $3,579 lower for patients receiving shorter-term therapy when compared with controls. By improving cognitive and daily functioning, donepezil may lower costs by improving medical management. PMID- 11925683 TI - Speech recognition solves only some problems of text-based health records. PMID- 11925684 TI - Speech recognition: how good is good enough? AB - Since its infancy in the early 1990s, the technology of speech recognition has undergone a rapid evolution. Not only has the reliability of the programming improved dramatically, the return on investment has become increasingly compelling. The author describes some of the latest health care applications of speech-recognition technology, and how the next advances will be made in this area. PMID- 11925686 TI - Organizational ethics and institutional integrity. PMID- 11925687 TI - Organization ethics or compliance: which will articulate values for the United States' healthcare system? AB - There have been two major stimuli leading to the institution of compliance/ethics programs. One has been Justice Department attention and the need for a government approved compliance program. The other has been the JCAHO's promulgation of standards addressing organization ethics issues. Many healthcare organizations have seen the requirements imposed on them by JCAHO (and the necessity to have a mechanism to address federal compliance as separate issues) and have developed two compartmentalized programs. But the intensity with which the government has pursued and will continue to pursue its "fraud and abuse" program means that most healthcare organizations now support and encourage the institution of compliance programs, while allowing organization ethics initiatives to become marginalized within the organization. This approach may ignore consideration of the larger issue of what each of these activities means for a healthcare organization that is attempting to define itself based on its stated values. Although using two separate groups to focus on these particular issues may fulfill the letter of the law for both the Federal Government and the JCAHO, we believe attention to these issues is better accomplished under the umbrella of a single committee. This pathway for attention to accreditation and compliance issues within an HCO is one that may serve the HCO better than two committees working on similar issues but with separate agendas. Nothing here should be construed as a suggestion that the goal of developing a comprehensive values-oriented organization ethics program should be abandoned. Instead, we have offered an assessment of the current environment and current direction of the healthcare organization--an assessment that leads us to the conclusion that instituting organization ethics programs that are not linked to or incorporated somehow within compliance programs will probably fail. This is a frustrating position for those who believe that compliance programs may ultimately undermine the goals of an organization ethics program. Nevertheless, it is important to realize that the legal compliance of an HCO, like its compliance to high standards of ethics, are considerations that determine the ethical climate of an organization. In the absence of a consensus concerning the values that should inform the larger healthcare system, it may be the only consideration that all healthcare organization stakeholders can agree upon, and so may represent the only stable and consistent platform ethically to evaluate the activities of the healthcare organization at the present time. This can be the basis for each HCO to develop its values-oriented ethical program which will define and support the organization's ethical climate for the organization, its staff, its patients, and its community. PMID- 11925688 TI - Ethics committees identify success factors: a national survey. PMID- 11925689 TI - Ethical concerns of staff in a rehabilitation center. PMID- 11925690 TI - Moving beyond "on the job training": preparing hospital ethics consultants for intensive care unit (ICU) rounds. PMID- 11925691 TI - Is "failure to thrive" syndrome relevant to Lithuanian healthcare ethics committees? PMID- 11925692 TI - Water: a clear and present danger. PMID- 11925693 TI - Drinking water supply and management practices in British Columbia, 1997-98. AB - BACKGROUND: Questions regarding control over a water fluoridation system in a British Columbia (BC) community led to a drinking water management survey in 1997 98. METHOD: A questionnaire was constructed based on published drinking water control and management criteria and was sent to 91 communities. RESULTS: 73/91 surveys were returned (80% response rate); 31 reported a protected water supply system, 40 reported logging and/or cattle grazing in watershed areas, 25 reported a lack of primary disinfection. Water fluoridation was reported in 12 locations, 3 of which did not monitor fluoride levels. Testing for protozoans was done routinely in 19 locations, 15 using approved laboratories. 15 water contamination events were reported, 9 of biological origin. Statistically significant associations were found between contamination events and: wooden delivery systems, lack of primary chlorination, increased number of protozoan tests, and increased number of personnel. INTERPRETATION: At the time of the survey in British Columbia, a number of communities were vulnerable to preventable drinking water contamination. PMID- 11925694 TI - Building capacity in evidence-based public health practice. PMID- 11925696 TI - The safety of Canadian early discharge guidelines. Effects of discharge timing on readmission in the first year post-discharge and exclusive breastfeeding to four months. AB - BACKGROUND: Few studies have examined earlier discharge in relation to Canadian guidelines for earlier discharge and infant feeding. We addressed differences in readmission (1 year post-discharge) and exclusive breastfeeding (4 months) for newborns and mothers discharged within 48 hours compared to those with a longer hospital stay. METHOD: A cohort of 1,357 vaginally delivered singleton normal newborns and their mothers (births between January 1, 1996 and March 31, 1997) were studied by linking five databases and a chart audit. RESULTS: Overall there were no differences in infant and maternal readmission or rates of exclusive breastfeeding. CONCLUSION: Canadian guidelines for earlier discharge appear appropriate for vaginally delivered singleton normal newborns and their mothers with timely home visitation. PMID- 11925695 TI - The effects of aerial spraying with Bacillus thuringiensis Kurstaki on children with asthma. AB - OBJECTIVE: To determine if aerially spraying a biological pesticide was associated with an increase in the symptoms or change in the Peak Expiratory Flow Rate of children with asthma. METHODS: A pre/post matched pairs cohort design was used. Children living in the spray zone were matched with children outside of the spray zone. Peak Expiratory Flow Rates, asthma symptoms and non-asthma symptoms were recorded in diaries. RESULTS: There were no differences in asthma symptom scores between subjects and controls, neither before nor after the spray; nor were there significant changes in Peak Expiratory Flow Rates for subjects after the spray period. CONCLUSIONS: No evidence of adverse effects from the use of the biological pesticide was found. We believe that this is the first paper to address the issue of whether or not aerial spraying with Btk has a harmful effect on children with asthma. PMID- 11925697 TI - The influence of prevalence and policy on the likelihood that a physician will offer HIV screening in pregnancy. AB - OBJECTIVE: To determine the extent to which provincial recommendations, reported regional prevalence rates and perceived local prevalence rates of HIV in pregnancy influence a physician's decision to routinely offer prenatal screening for HIV. DESIGN AND METHODS: A random sample of 5,052 family physicians and obstetricians were surveyed by mail. Logistic regression was used to explore the relationships among the variables of interest. RESULTS: The response rate was 61%. Of these, 69.2% provided prenatal care and were included in the analysis. Physicians were more likely to routinely offer HIV testing if they practiced in provinces with recommendations that supported the universal offer of a test (O.R. = 5.80), independent of living in a region with an estimated prevalence rate exceeding 5/10,000 (O.R. = 1.76), or the perception that the infection rate in their practice justified universal counselling of pregnant women (O.R. = 10.41). CONCLUSIONS: Provincial recommendations supporting universal HIV testing in pregnancy are reflected in physician practice. PMID- 11925698 TI - The affordability of a nutritious diet for households on welfare in Toronto. AB - OBJECTIVES: This study assesses the affordability of a nutritious diet for households in Toronto that are supported by welfare. METHODS: For three hypothetical households, welfare incomes were compared to the monthly costs for food, shelter, and other essential expenditures in Toronto. RESULTS: If households lived in market rental accommodation, average monthly incomes were insufficient to cover expenses for the single-person household and two-parent family, and barely adequate for the single-parent family considered in this study. However, the single-parent family's actual income fell below expenses for six months of the year. For households with children, the relative inadequacy of welfare increased as children grew older. Living in rent-geared-to-income housing afforded substantial financial advantage, but the welfare income of single-person households was still insufficient to meet basic needs. INTERPRETATION: These findings indicate discrepancies between welfare incomes and costs of basic needs, which may explain the vulnerability of welfare recipients to food insecurity. PMID- 11925699 TI - Care-giving and care-seeking behaviours of parents who take their children to an emergency department for non-urgent care. AB - OBJECTIVE: This study explored the care-giving and care-seeking behaviours of parents who took their children to the emergency department (ED) of a large urban hospital in Western Canada for non-urgent care. METHOD: Data were collected from a convenience sample of 114 parents during a two-week period in January 1992, using a self-administered 53-item questionnaire. The data were analyzed using nonparametric statistics (Chi-square) and thematic content analysis. RESULTS: Eighty-two percent of parents were unsure of or overestimated the seriousness of their child's illness or injury. Prior to attending the ED, 40% of parents administered comfort measures and 45% gave medications. Only 17% of parents used at-home reading materials, while 31% sought lay advice. Fifty-eight percent of parents did not try to contact their family physician or the ED by telephone before coming to the ED. Forty-eight percent of parents who phoned their family physician were unable to obtain advice, and those who did were almost always referred to the ED. Eighty-eight percent of those who phoned the ED were instructed to bring the child to the ED. INTERPRETATION: The results suggest the need for more acceptable, accessible community primary care services. PMID- 11925700 TI - Teen sexuality. Reaching out in the malls. AB - BACKGROUND: Existing sexual health programs have not significantly reduced teen pregnancies or sexually transmitted diseases. A more creative approach is needed. METHODS: An assessment of 539 teens in one Ontario city was conducted to identify knowledge about and use of birth control, comfort in discussing sexual health, and preferred sites, providers and methods of service delivery. RESULTS: Knowledge of, and comfort discussing, birth control was not associated with frequency of use but was associated with grade. Adolescents were less comfortable discussing sexual health with teachers than health professionals. Over time, comfort increased with health professionals, but not teachers. Sexually active teens reported willingness to attend mall-based clinics. CONCLUSIONS: Using birth control appears to be maturational given its association with grade. Since teens were consistently less comfortable with teachers, providing sexual health services in schools is likely ineffective. Teens may respond to clinics in creative settings such as malls. PMID- 11925701 TI - Seroprevalence and risk factors for hepatitis A among Montreal street youth. AB - OBJECTIVE: To estimate the prevalence of hepatitis A virus (HAV) antibodies among Montreal street youth. METHOD: Anti-HAV antibody testing was performed on blood samples from a hepatitis B and C study conducted among street youth in 1995-96. RESULTS: Among the 427 youth aged 14 to 25 years, prevalence of HAV antibodies was 4.7% (95% confidence interval [CI]: 2.9%-7.2%). A multivariate logistic regression analysis showed that birth in a country with a high anti-HAV prevalence (Adjusted odds ratio [AOR]: 200.7; 95% CI: 38.1-1058.4), having had sexual partner(s) with history of unspecified hepatitis (AOR: 13.8; 95% CI: 4.2 45.2), and insertive anal penetration (AOR: 5.1; 95% CI: 1.6-16.7) were independently associated with infection. CONCLUSION: Based on the relatively low HAV prevalence, the high prevalence of risk factors for infection, and the substantial hepatitis B and C prevalence, vaccination against hepatitis A is now actively promoted among Montreal street youth. PMID- 11925702 TI - [Knowledge and perceptions of adolescents regarding oral health]. AB - OBJECTIVE: The objective of this study was to measure knowledge and perception related to oral health in Quebec children. METHOD: For this study, a provincial sample of Quebec adolescents, 13 to 14 years old, was set up. Adolescents answered a 46-item questionnaire, specifically designed for the study. The questionnaire was completed by over 1,300 adolescents (participation rate = 53.0%). RESULTS: The answers indicate that adolescents know the importance of toothbrushing and dental services utilization for dental caries prevention. However, their knowledge about dental sealant and fluoride is inadequate. Teenagers believe that they have a significant role to play in the prevention of dental diseases. However, many adolescents consider tooth loss to be a normal consequence of age. The majority of adolescents report their oral health as good. It is noted that dental aesthetics is a more significant factor for them than the quality of their teeth. Finally, dental care is a source of anxiety in one third of the adolescents. CONCLUSION: The study of teenagers' knowledge and perceptions related to oral health shows that these elements are strongly influenced by environment, norm and culture. To improve dental health concerns in teenagers, public health policies should concentrate on these elements. PMID- 11925703 TI - Correlates of suicide attempts in an open cohort of young men who have sex with men. AB - OBJECTIVE: To investigate baseline correlates of attempted suicide in a large cohort of young gay and bisexual men. METHODS: Participants completed annual questionnaires asking demographic information, sexual behaviours, history of forced and paid sex, comfort with sexual orientation, use of illicit drugs, and validated measures of depression, social support, alcohol dependency, self-esteem and suicide ideation and attempts. Contingency table analysis and step-wise logistic regression were used to identify potential predictors of self-reported suicide attempts. RESULTS: Of 345 gay and bisexual men eligible for this cross sectional analysis, 150 (43.5%) reported that they had ever considered suicide and 67 (19.4%) that they had attempted suicide at least once. After adjustment for multiple explanatory variables, the use of nitrite inhalants (poppers) (AOR = 2.37; 95% CI 1.30, 4.33), social support scores below the 75th percentile of all scores (AOR = 2.19; 95% CI 1.18, 4.09) and low or moderate self-esteem (AOR = 3.73; 95% CI 2.03, 6.86) were independently associated with elevated risk of attempted suicide. CONCLUSION: Our data indicate that men in this analysis who ideate or attempt suicide earlier in life are more likely to report lower social support and self-esteem, and high popper use. PMID- 11925704 TI - Ask and ye shall plan. A health needs assessment of a university population. AB - BACKGROUND: In the development process of establishing a Campus Health Resource Centre, a health needs assessment of 691 students was conducted at the University of Manitoba. METHODS: Students were surveyed by their peers to identify the health education needs of this population. The process of the health needs assessment is described and the results have formed the basis for a range of programs and services offered on campus. RESULTS: Students showed interest in learning about stress management, cold and flu prevention, ergonomics and lifestyle (exercise, nutrition) issues. CONCLUSION: Of note is low interest in topics generally thought to be important to students such as contraception, safer sex, and STD/AIDS prevention. PMID- 11925705 TI - [Awareness of the Info-Sante CLSC telephone service by users of urgent care services]. AB - BACKGROUND: The CLSC's Info-Sante telephone service is one of the Quebec health network's reorganization measures aimed at decreasing the number of visits to hospital emergency rooms for non-urgent health problems. This study analyzes the awareness of this service among users of hospital emergency services and walk-in medical clinics. METHODS: In all, 850 patients were interviewed. Logistic regressions were performed. RESULTS: The results revealed that 75.5% (CI .95 = 0.73-0.78) of the participants were indeed aware of the CLSC's Info-Sante telephone service. Awareness was found to be dependent on age, sex, income, the use of walk-in CLSC medical services, the availability of a regular source of health care and the perception of one's health status. INTERPRETATION: Further strategies should be developed in order to increase the use of this service, thus helping to guide patients to more appropriate sources of health care. PMID- 11925706 TI - [Beneficial effect of maternity leave on delivery]. AB - OBJECTIVE: To identify the contribution of the duration of the prenatal maternity leave on term delivery. METHOD: Characteristics of the prenatal maternity leave and delivery among 363 working women who had delivered a full-term infant at 1 of 4 hospitals in Montreal during 1996 were studied. RESULTS: The presence of an intervention or complication during delivery was observed in 68.9% of the participants. The average duration of the prenatal maternity leave was about 8 weeks (SD = 7). The adjusted risk of a difficult delivery decreased significantly with the duration of the prenatal maternity leave (OR = 0.96; 95% CI: 0.93-0.99). CONCLUSION: The duration of the maternity leave before delivery is associated with an easier term delivery for working women. PMID- 11925708 TI - [Traumatology and military surgery at the change from the Middle Ages to modern times]. PMID- 11925707 TI - The art and science of evidence-based decision-making ... epidemiology can help! PMID- 11925709 TI - [Adam Christian Thebesius (1686-1732) and the discovery of the Vasa Cordis Minima]. AB - In first years of the 18th century, Raymond Vieussens (1635-1713) and Adam Christian Thebesius (1686-1732) discovered the Vasa cordis minima, today known as Vasa Thebesii; in 1868, Odilon Marc Lannelongue (1840-1911) described the so called crypts of Lannelongue in the right atrium. The present study gives a historical survey over the research on the cardiac vascularization before 1700 and presents these three authors and their works on the Thebesian vessels with a biographical introduction and a commented translation. As for the biography of the Silesian anatomist Thebesius large, mostly inedited materials from Polish archives and from family archives were studied, among them, poems, a handwritten biography and a pedigree of the family. A schedule tables the studies and articles on the Thebesian vessels and the Thebesian valve published since 1708. As a commentary to the bibliography, the present stage of knowledge on the Thebesian vessels is shortly summarized. Although a compensatory function of the Thebesian vessels in coronary sclerosis and other pathological conditions has been discussed repeatedly, methodical difficulties still do not allow a reliable judgement on the role of the Thebesian vessels in physiological and pathological conditions. This study shows the contradictory results and summarizes all statements which have been published on the embryology, comparative anatomy, macroscopical and (electron-)microscopical anatomy, physiology, pathophysiology and cardio-surgery. The aim is to encourage an interdiscplinary discussion on the base of the thorough literature study. PMID- 11925710 TI - Two outbreaks of botulism associated with fermented salmon roe--British Columbia, August 2001. PMID- 11925711 TI - [Sexuality and pregnancy in adolescence: between nature and culture]. PMID- 11925712 TI - [Epilepsy and mysticism]. PMID- 11925714 TI - [Genetics of neuroendocrine tumors]. AB - Genetics of neuroendocrine tumours is mainly related to major inherited syndromes which predispose to the development of tumours in endocrine glands. Clinical and genetic studies on multiple endocrine neoplasia (MEN) syndromes show the diversity and complexity of cellular and genetic pathways involved in endocrine tumor genesis. Nevertheless, we might expect that recent cloning of most of the genes involved in MEN and related syndromes, has been a powerful and historical step to help clinicians in differential diagnosis of MEN and related diseases, mainly in cases which show familial association of various types of endocrine tumors. To date, 4 to 6 genes may be commonly analyzed and a single patient diagnosed for a specific syndrome, thus leading to a better clinical and therapeutical follow-up for patients and related predisposed individuals. PMID- 11925713 TI - [Definition and histologic characterization of neuroendocrine tumors]. AB - Neuroendocrine tumours are defined by a common phenotype, which is not supported by a common embryologic origin. This common phenotype is characterized by the expression of general neuroendocrine markers, and sometimes by cell specific hormonal products. Neuroendocrine tumours are ubiquitous, but the major localizations are the digestive tract. According to the classification of lung tumours, they are divided in low grade and high grade tumours. Since most digestive and pancreatic tumours are low grade tumours, a specific classification of neuroendocrine tumours was recently proposed. Size, degree of invasion, major secretion, and proliferation rate are the main criteria of this classification. Most neuroendocrine tumours are sporadic. A few cases occur in the context of a multiple endocrine neoplasia type 1. PMID- 11925715 TI - [Clinical spectrum of digestive neuroendocrine tumors]. AB - Digestive neuro-endocrine tumours have a broad and initially misleading clinical spectrum. Tumours from the duodenopancreatic area should be distinguished from digestive carcinoid tumours. In the first group, insulinomas, gastrinomas, and non-functioning tumours are the most frequent. Insulinoma is responsible for hypoglycaemic symptoms (coma, confusion, seizure, psychiatric disorders) associated with adrenergic response (sweat, tachycardia, palpitations). Gastrinoma is responsible for the Zollinger-Ellison syndrome, which associates peptic ulcers in the oesophagus, stomach, and duodenum without Helicobacter pylori infection, and chronic volumogenic diarrhoea. Non-functioning tumours are recognised fortuitously or at a late stage, when large tumour mass contrasts with often unaltered general condition. Carcinoid tumours are mainly located in the appendix, the rectum, and the small bowel. In the 2 first conditions, the diagnosis is most often made on a resection specimen after uneventful appendectomy or polypectomy; in the latter, the carcinoid syndrome is frequent, combining cutaneous flushing, motor diarrhoea, tricuspid valve insufficiency and bronchospasm. PMID- 11925716 TI - [Biology and imaging of neuroendocrine tumors]. AB - Endocrine tumours constitute a network of tumours scattered in the body and characterized by several common features including their capacity to secrete hormones, their association as part of inherited syndrome and their ability to be explored by functioning imaging. We review main advances in the biological and morphological work-up of these rare tumours, including the role of chromogranine A measurement and somatostatin receptor scintigraphy. PMID- 11925717 TI - [Locoregional therapy of neuroendocrine tumors]. AB - In digestive neuroendocrine tumours, surgery is the cornerstone of the treatment of the primary tumour. The diameter of the lesion is the main prognostic indicator and consequently impacts the extent of the resection. Types of resection, regarding to tumours sizes and locations, are reported. In metastatic forms, an aggressive policy of multidisciplinary treatments is proposed. Arterial chemoembolization is very efficient in controlling clinical symptoms and liver tumours progression, and allows secondary radical resections in selected cases. Chemoembolization is actually considered as the first line treatment in well differentiated forms, with rapid progression. PMID- 11925718 TI - [Biotherapy of neuroendocrine tumors]. AB - The somatostatin analogues have an excellent symptomatic therapeutic effect (60 90% responses in carcinoid syndrome) which permits an amelioration in the quality of life, particularly in patients with metastatic tumours. The availability of long-acting somatostatin analogues assuring stable plasma concentrations over several weeks (thanks to progressive liberation) further facilitates the use of these medications by allowing a reduction in the number of injections. Interferon also has an anti-secretory effect, albeit inferior to that of the somatostatin analogues, but potentially of interest in the case of resistance to these products. PMID- 11925719 TI - [Chemotherapy of neuroendocrine tumors]. AB - Chemotherapy has few impact on neuroendocrine tumour patients outcome: it may decrease functional secretory symptoms and increase slightly median survival time. In islet-cell carcinoma of the pancreas the standard chemotherapy regimen is the combination of streptozotocin and doxorubicin. It induces 50% response rate and may increase by 50% the median survival. In enterochromaffin-cell tumours chemotherapy has modest impact and the combination of 5-fluorouracile and streptozotocin is the standard regimen. In undifferentiated enterochromaffin-cell tumours of unknown primary the standard chemotherapy regimen is the combination of etoposide and cisplatin which induces 50% response rate with probably no impact on overall survival. Chemotherapy must be indicated within the frame of a multidisciplinary approach and only in patients who have refractory and progressive disease. Indication of chemotherapy must be balanced with indications of biotherapies and embolization. PMID- 11925721 TI - [Presidential debate]. PMID- 11925720 TI - [Treatment strategy of neuroendocrine tumors]. AB - Therapeutic strategy of neuroendocrine tumours is complex, due to their heterogeneity and to the fact that although generally slow growing, a significant proportion demonstrates aggressive tumour growth. Symptomatic carcinoid syndrome and various pancreatic endocrine tumours with symptomatic syndromes are well controlled with somatostatin analogues. Surgery remains the mainstay of treatment if the tumour can be resected. Metastatic pancreatic neuroendocrine tumour are treated when resection is not feasible with combination chemotherapy using adriamycin and streptozotocin, which remains a standard of care. In well differentiated tumour of the gut or the lung there is no clear standard of chemotherapy and treatment vary according to the tumour course. In indolent cases, somatostatin analogues are the best treatment, in case of aggressive tumours chemoembolisation should be preferred when the disease is located or predominant in the liver. Poorly differentiated tumours are treated by combination chemotherapy with etoposide and cisplatin, and surgery has no indication. Gastrinoma and other pancreatic tumours arising in the context of multiple endocrine neoplasia type I disease need a specific therapeutic strategy. PMID- 11925722 TI - [Non-traumatic coma]. PMID- 11925723 TI - [Ovarian tumors]. PMID- 11925724 TI - [Psychomotor development of infants and children. Normal and pathologic aspects (sleep, digestion, sphincter control, psychomotor, language, intelligence). The early role of the mother-child relationship and its importance in learning disorders]. PMID- 11925725 TI - [Acute myeloblastic leukemia]. PMID- 11925727 TI - Engaging physicians in the accreditation process. PMID- 11925726 TI - [Arteriosclerosis obliterans of the aorta and lower limbs. Aneurysms]. PMID- 11925729 TI - ORYX pricing model for performance measurement systems approved for 2003. PMID- 11925728 TI - Identifying and avoiding potential hazards in the laboratory. PMID- 11925730 TI - Improving the medication use process. PMID- 11925731 TI - Mechanical stress as cause of aortic valve disease. Presentation of a new aortic root prosthesis. AB - We present an overview of studies on the aortic valve and propose that mechanical stress is a main causative factor in the degenerative valvular disease. In the normal aortic valve, the leaflets have a smooth surface, free of wrinkles and creases, throughout the opening process. This smooth leaflet surface during motion is achieved by the "pull and release" movement of the commissures, which occurs because of the compliance of the aortic root. When the aortic root is stiffened, either by artificial means or by the loss of elasticity due to aging, the leaflet dynamics change significantly. The leaflets develop a significant number of creases and wrinkles during the opening process. In the bileaflet valve, the leaflets develop similar creasing and wrinkling during the opening process. This happens mainly due to the less-than-ideal design of the bileaflet valve and in spite of the compliant aortic root. When the aortic valve is spared using a noncompliant tube graft, a similar phenomenon of leaflet creasing occurs. Because the creasing produces high stresses from bending and buckling, it is damaging to the leaflet tissue and can lead to degenerative and calcific valvular disease. Based on these observations a new aortic root prosthesis with compliant sinuses has been designed for the valve sparing operation. PMID- 11925733 TI - Patient's opinion about written information before laparoscopy: a consecutive series of 100 cases. AB - PURPOSE: The aim of this study was to assess the opinion of the surgical patient concerning written information before laparoscopic operations. MATERIAL AND METHODS: A prospective and consecutive series of 100 patients was studied. Information sheets concerning the planned laparoscopic operation were distributed at preoperative consultation. A short and clear questionnaire regarding this information was answered upon hospitalization. RESULTS: The majority of the 87 patients, who had read the information, were very pleased to be informed about the techniques (91%) and the risks (97%), although a significant group was worried by the explanations of the risks (41%). None of the patients cancelled the planned operation. None of the patients had become less confident in the surgeon. Most of the patients (95%) found this system of informed consent necessary. CONCLUSIONS: Surgeons should no longer be reluctant to distribute standardized information sheets, as a majority of patients find this system of information necessary. PMID- 11925732 TI - Spontaneous pneumothorax: needle aspiration or chest drain? Evidence-based medicine. PMID- 11925734 TI - Filling sensations after restorative proctocolectomy. AB - OBJECTIVE: In order to improve insights in rectal filling sensation, we studied pouch filling sensations after ileal J pouch-anal anastomosis (IPAA) before and after re-establishment of bowel continuity. METHODS: Anal manometry and a pouch filling sensation test were performed before as well as 1 and 6 weeks after closure of the loop ileostomy in 17 patients who had undergone restorative proctocolectomy with stapled (8 patients) or manual pouch-anal anastomosis (9 patients). The results were compared with those of 12 control subjects. RESULTS: Before ileostomy closure, pouch pressure necessary for inducing the respective sensation thresholds was higher than in controls; the difference was significant for constant and urge sensation. The volumes for urge and maximum tolerable sensation level were significantly lower, with reduced pouch compliance. After stoma closure, pressure and volume thresholds at all sensation levels became completely comparable with control data. No relevant differences were observed between stapled and manual ileal pouch-anal anastomoses. CONCLUSIONS: All levels of filling sensation levels are preserved after restorative proctocolectomy and their parameters are comparable with those of normal rectal filling sensation. Diversion of an ileal J pouch results in resetting of filling sensation thresholds towards lower volume and higher pressure values, but all sensation thresholds normalize within 6 weeks after stoma closure. These data document that neither the rectum, nor the mucosa of the anorectal junction and upper part of the anal canal are involved in filling and urge sensation. PMID- 11925735 TI - Primary anastomosis and diverting colostomy in diffuse diverticular peritonitis. AB - BACKGROUND: Despite the well documented morbidity associated with its reversal, Hartmann's procedure remains the favoured option in patients with complicated diverticular disease in the presence of diffuse peritonitis. A prospective study was conducted to determine whether primary anastomosis with diverting colostomy constitutes a valid alternative to the Hartmann procedure. METHODS: Between 1994 and 1998, all patients with diffuse peritonitis due to perforated diverticulitis of sigmoid origin underwent resection and primary anastomosis with diverting colostomy. Restoration of colonic continuity was programmed six weeks later, after verification of the anastomose by gastrografin enema. The group included 5 men and 15 women with a mean age of 72 years (32-97 years). The ASA classification of the patients was as follows: ASA II (n = 2), ASA III (n = 12), ASA IV (n = 3), ASA V (n = 3). The mean delay between onset of symptoms and surgery was 74 hours (8-215 hours). RESULTS: Operative mortality and morbidity was 15% (n = 3) and 50% respectively. No patients showed signs of suture disruption and this was confirmed by routine radiological controls of the anastomoses. Mean length of hospitalization was 20 +/- 10 days (SD; median: 18 days). Closure of the colostomy using a small peristomal incision was performed in all surviving patients after a mean delay of 45 +/- 9 days (range 28-67 days). Mean length of hospitalization for colostomy closure was 7 +/- 3 days (range 3-18 days) without mortality. CONCLUSIONS: Applied systematically to all patients with diffuse peritonitis due to perforated diverticular disease, primary anastomosis was found to be as safe as the Hartmann procedure but appears to be superior in terms of total length of hospital stay, interval to stoma closure and rates of stoma closure. Primary anastomosis with diverting colostomy could constitute a valid alternative to the Hartmann procedure in selected patients with complicated diverticular disease, even in the presence of diffuse peritonitis. PMID- 11925736 TI - Primary mechanical stapled anastomosis in surgery for colorectal emergencies. AB - From May 91 to March 99 a consecutive series of 100 acute obstructions or perforations of the left colon or rectum were treated by primary resection with mechanical anastomosis using a double or triple stapling technique without proximal colostomy. There were 8 postoperative deaths (8%) due to sepsis, acute respiratory distress syndrome, pulmonary embolism, stroke, and cachexy. Complications occurred in 29% of surviving patients. Clinical anastomotic leaks were observed in 7%, respiratory infection in 8%, wound infection in 8% and major cardiovascular problems in 4% of patients. The median hospital stay was 19 days. The morbidity and mortality of this series did not exceed the cumulative morbidity and mortality that can be expected after staged surgery. Compared with staged surgery, immediate resection and anastomosis using an entirely mechanical suture, thereby avoiding the problems of colostomy and reducing the length of hospital stay, has significant advantages for patients. PMID- 11925737 TI - Effects of neurotensin on the healing of experimental anastomosis of the colon. AB - The leakage of colonic anastomoses increases perioperative morbidity significantly. The purpose of the study was to investigate the influence of neurotensin, an intestinal trophic peptide, on the healing of colonic anastomosis. Forty-two Wistar-albino rats were separated into three equal groups: Group 1 (control group)--segmental resection of the left colon and end-to-end anastomosis; Group 2 (dexamethasone group)--resection and anastomosis, plus 0.1 mg/kg/day of dexamethasone; Group 3 (neurotensin group)--same surgical procedure plus 300 micrograms/kg/day of neurotensin. Bursting pressure and tissue hydroxyproline content were determined as parameters of the anastomosis strength and healing on the 3rd and 7th days postoperatively. On the 3rd day, mean bursting pressures were 141.4, 146.7 and 73.1 (p = 0.0001) cm of water in the control group, dexamethasone and neurotensin groups respectively. On the 7th day, bursting pressures were measured as 237.4, 100.6 (p = 0.0001) and 72.7 (p = 10( 6)) cm of water, in the control group, dexamethasone and neurotensin groups respectively. Between the 3rd and 7th days, bursting pressures were increased significantly in the control group (p = 0.0001), decreased in the dexamethasone (p = 0.048), and maintained their lowest values in the neurotensin (p = 0.96) groups. On the 7th day, mean hydroxyproline levels were measured as 9.20, 3.30 (p = 0.007), 2.86 (p = 0.007) micrograms, in the control group, dexamethasone, and neurotensin groups respectively. Between the 3rd and 7th days, tissue hydroxyproline levels were increased significantly in the control group (p = 0.004), decreased in the dexamethasone (p = 0.03), and maintained their lowest values in the neurotensin (p = 0.87) groups. The anastomosis resistance to intraluminal pressure was weak, tissue collagen content was insufficient, and healing was inadequate in the dexamethasone and neurotensin groups in respect to the control group. We concluded that neurotensin impaired the healing, and affected the strength of the colonic anastomosis. PMID- 11925738 TI - A newly designed vertebral replacement implant to reconstruct the thoracolumbar spine anteriorly. AB - Calcium phosphate compounds are becoming of increasingly great importance in the field of biomaterials and, in particular, as bone substitutes. In this way donor side morbidity can be avoided. A type A lesion of the vertebral body can be the reason of a neurological deficit, requiring a corpectomy by means of an anterior approach, tricortical bone grafting and internal stabilization to get a fusion over time. It is our aim to introduce in this study a newly designed vertebral replacement device, tested in an animal model, using domestic pigs. Two biomaterials, Cementek and Biobon were primarily macroscopically and microscopically evaluated, using a critically sized bone defect of a vertebral body and compared with autologous bone grafts. The final tests consisted of two groups of four animals. A corpectomy at the level of L4 was created and stabilized by means of the newly designed vertebral replacement implant. Autologous bone grafts were applied in one group of animals and Cementek in the other to fill the remaining space. Fusion was studied using anteroposterior and lateral radiographs, followed by a computer tomography. The biocompatibility of the biomaterials and autologous bone grafts were in order of: Cementek > autologous bone grafts > Biobon. In the final tests after a six months period, a fusion was diagnosed in four cases, two delayed unions, and two obvious non unions. An analysis of the failures in this animal study can possibly clarify the shortcomings of this concept. Perhaps, subtle changes of the design combined with other synthetic bone substitutes can possibly improve the fusion rate in the near future. PMID- 11925739 TI - Transverse fracture-dislocation of the sacrum: a diagnostic pitfall and a surgical challenge. AB - Transverse fracture-dislocations of the sacrum are rare. Associated lesions of the lumbosacral spine as well as neurological injuries are common. Conventional radiographs of the pelvis often fail to clearly visualize the fracture. Delayed diagnosis increases the risk of progressive neurological disfunction. True lateral sacral views and CT-scans with 3-dimensional reconstructions are very helpful in establishing the full extent of the injury. These examinations should be considered in all patients with a history of high energy trauma and clinical signs indicating lumbosacral injury, such as severe low back pain and neurological disturbances of the lower extremities. The management of transverse sacral fracture-dislocations with or without associated neurological damage is controversial. Conservative treatment is associated with a high rate of persistent deformity and residual neurological dysfunction. Surgical management allows for anatomical fracture reduction, stable fixation and revision of the spinal canal and lumbosacral nerve roots. The dorsal approach is preferred. Two patients with transverse sacral fracture-dislocations and neurological disturbances are presented. One patient had an additional fracture-dislocation of the lumbar spine at the L4L5 level with intrusion of the lumbosacral spine into the pelvis. Both lesions in this patient were successfully stabilized using an internal fixator system. The other patient presented with a bilateral transforaminal sacral fracture. The transverse component was not recognized on the initial radiographs, which resulted in loss of reduction and progressive neurological disfunction after sacroiliac screw fixation. PMID- 11925740 TI - Solid and cystic tumor of the pancreas: a case report. AB - A case of a solid and cystic tumor of the pancreas occurring in a 15 year-old girl is reported. This patient was admitted with epigastric and left upper quadrant pain and vomiting. A mass was detected in the pancreas on CT scan and the patient was referred for surgery. A distal pancreatectomy was performed. A pathological diagnosis of solid and cystic tumor of the pancreas was made and the patient was discharged uneventfully. PMID- 11925741 TI - Enucleation of a giant hepatic hemangioma in a Jehovah's witness. PMID- 11925742 TI - Typhlitis in an immunocompromised patient. AB - We report the case of a young man who presented with right lower quadrant pain, while being treated for acute lymphatic leukaemia. Clinical findings were suggestive of appendicitis. Computed tomography scan of the abdomen was done and showed the presence of a neutropenic enterocolitis or typhlitis. PMID- 11925743 TI - Successful iterative drainage and partial hepatectomy for pyogenic liver abscess in a HIV seropositive patient. AB - The case of cryptogenic Escherichia coli pyogenic liver abscess in a 59-year-old Human Immunodeficiency Virus (HIV) seropositive man is reported. The initial treatment was a percutaneous drainage. As the abscess did not reduce in size, surgical drainage was planned but during surgery a necrosectomy had to be performed resulting in a partial hepatectomy. After nine months of amoxicillin clavulanic acid treatment, drainage and highly active antiretroviral therapy, the patient recovered completely. It is expected that because of highly active antiretroviral therapy, mortality rates of surgical interventions in patients with HIV infection will decrease. Because of the increased life expectancy in persons with HIV infection, the criteria for considering surgical interventions in these patients should be broadened. PMID- 11925744 TI - Benign mucinous cystadenoma of the appendix. Report of 3 cases. Review of the literature. PMID- 11925745 TI - Aortic graft infection. What have we learned during the last decades? PMID- 11925746 TI - [The role of reactive oxygen species as signal molecules in tissue metabolism in oxidative stress]. AB - This review deals with literature data on the functioning of the reactive oxygen species (ROS) as second messengers. ROS induce various biological processes such as stimulation of protein phosphorylation, Ca(2+)-signaling, phospholipid hydrolysis and transcription factor activation. Physiological significance of the role of biological oxidants in the regulation of signal transduction and the mechanisms of the adaptation of organism to extremital conditions are discussed. Oxidative stress is a common step for the development of many diseases characterized by the intensive ROS generation. PMID- 11925747 TI - [Brain monoamine oxidase in prematurely aging OXYS rats]. AB - Rats OXYS with manifestations of genetically determined premature aging and OXYR free from these manifestation were studied. Eight month old OXYS and OXYR rats were characterized by similar brain stem MAO A and MAO B activities and their control at the transcription level. In hemispheres of 8 month old OXYS rats MAO A activity was higher than in OXYR. Emotional stress did not affect either MAO A or MAO B in brain stem of the two strains, and lowered MAO A activity in hemispheres of OXYS rats. PMID- 11925748 TI - [Cytotoxic effect of conjugates of L-lysine-alpha-oxidase with monoclonal antibodies on human tumor cells in vitro]. AB - The conjugates of L-lysine alpha-oxidase and monoclonal antibodies ICO-80 towards CD-5 receptor were produced using glutaraldehyde. The cytotoxic effect of conjugates on Yurkat cells line appeared to be lower in comparison with the native enzyme. Negligible decrease of conjugate biological activity may be explained by the large molecular weight of conjugate, which is several times higher than the molecular weight of the native enzyme. Such conjugates can not penetrate into the cells. So they catalyze the hydrogen peroxide formation, the main damaging agent, probably only outside the cells. We suppose also that the free native enzyme penetrates into the cell and activates there the oxidative deamination of L-lysine and correspondingly the hydrogen peroxide formation. This may be the proper explanation for the higher cytotoxic effect of L-lysine alpha oxidase on Yurkat cell line. PMID- 11925749 TI - [Effect of plant preparations on lipid peroxidation parameters in acute toxic hepatitis]. AB - The influence of the original vegetable complexes (which include: juices of beet rout and carrot, decoction of degrose berries, extracts of corn silk, leaves of peppermint and some other components) on the indicators of the cytolysis, lipid peroxidation and antioxidant system of serum of the laboratory rats with acute toxic hepatitis, was investigated. The vegetable complexes exhibited antioxidant effects which were proved by the reduction of the final and intermediate products of lipoperoxidization, the absence of decline of the level of endogenous alpha tocopherol content and glutathione dependent enzymes. PMID- 11925751 TI - [The use of empirical correlations in adenine nucleotide systems for prediction of AMP level in the brain in hypoxia]. AB - The empirical relationship between the changes of the AMP concentration and the AMP content under different kinds of hypoxia was showed. This relationship is determined by the function which is closely related to the totality parameters of the brain energy metabolism. This function may be used for the prognosis of the brain energy state under hypoxia. PMID- 11925750 TI - [Activities of the liver microsome reductases in adult and aged rats during stress]. AB - The influence of 0.01% sodium dodecyl sulphate, 1.5 and 6.0 M urea and 0.03 M hydrogen peroxide to the NAD(P)H: 2,6 dichlorphenolindophenol reductase activity in livers of adult and old Wistar rats during immobilizing stress was interested. Obtained results indicate that the NADPH--dependent reductase is more resistant to modulating effect of sodium dodecyl sulphate, hydrogen peroxide and urea than NADH-dependent enzyme. The significant decrease of NADH: 2.6 dichlorphenolindophenol reductase sensitivity to the action of all studied modulators occurs in old rats. The similar changes appears in the adult rats liver during stress. The old rats immobilization is accompanied by a decrease of this enzyme activity and the reduction of the influence of all studied modulators to NADH: 2.6 dichlorphenolindophenol reductase as compared with adult ones. These changes in the activity and properties of microsomal NADH: 2,6 dichlorphenolindophenol reductase promote more pronounced decrease of the substrate hydroxylation in the liver of old rats during stress compared to adult ones. PMID- 11925752 TI - [Cloning of the human cytochrome p450 2B6 gene in Escherichia coli cells]. AB - Human cytochrome P450 2B6 gene from plasmid pUC9, carrying cytochrome CYP2B6 cDNA was cloned into eucariotic expression vector pcDNA 3.1 (+). Cytochrome P450 2B6 gene in recombinant plasmid pcDNA 3.1 (+)/CYP 2B6 was expressed in E. coli cells. The expression of catalytically active recombinant protein was 60-100 nm per liter of the culture medium. PMID- 11925753 TI - [Changes in metabolism of sialoglycoproteins of the rat stomach during long-term immobilization stress and glucocorticoid hormone deficiency]. AB - The effect of glucocorticoid hormone deficiency (caused by administration of chlordithane) on the methabolism of gastric sialoglycoproteins at the prolonged immobilized stress was investigated in male albino rats. Under conditions of prolonged deficit of glucocorticoid hormones (> 10 days) the imobilised stress caused higher increase of catabolic processes of sialoglycoproteins than in rats with normal level of glucocorticoids. PMID- 11925754 TI - [Lipid peroxidation in the cyst fluid of craniopharyngiomas]. AB - Comparing of the craniopharyngiomas cyst fluid, collected during the surgery, with blood plasma revealed significantly enhanced lipid peroxidation and lactate accumulation in tumour cysts contents. These processes were usually more prominent in endosuprasellar than in suprasellar craniopharyngiomas. The maximum of free radical formation in conjunction with the lowest level of malondialdehyde (MDA) was observed in primary tumours with relatively high proliferation rate. Controversial parameters ratio characterised the recurrent craniopharyngiomas, and in cyst fluids of the primary tumours with low proliferation rate the medium parameter's values were observed. Thus, the estimation of free radical formation level and MDA concentration in cyst fluid of craniopharyngioma seems to be prognostic for the tumour recurrence. PMID- 11925755 TI - [Blood fatty acid transporters in acute pancreatitis]. AB - A relationship between of acute pancreatitis severity and concentrations of blood transporters of nonesterified fatty acids (albumin) and esterified fatty acids (lipoproteins) was studied. In necrotizing pancreatitis, there was a positive correlation between decrease of albumin and lipoprotein concentrations in serum, but no correlation was in edematous pancreatitis. High levels of serum albumin (> 32 g/L) and lipoproteins (cholesterol + triglycerides, CH + TG, > 260 mg/dL) had good prognostic value in terms of favourable development of this disease, whereas lower concentrations indicated unfavourable prognosis. Both albumin and lipoproteins appeared in the peritoneal exsudate where they represented 73 and 53% of the serum concentrations, respectively. Albumin efflux from blood to the exsudate correlated with lipoprotein efflux (r = 0.7) in necrotizing pancreatitis. The latter suggests that proteins penetrated together the tissue barrier blood/peritoneal space. The less albumin and lipoprotein concentrations in serum, the more they are in the exsudate (r = -0.57 and -0.47, respectively); probably, their concentration decrease in blood is due to to their efflux into the exsudate. PMID- 11925756 TI - [Modeling of monoamine oxidase B active site by computer moulding]. AB - Using previously developed approach for active site modelling of enzymes with unknown spatial structure (Veselovsky et al., Biochemistry (Moscow), 65, 1072 1079) substrate/inhibitor binding site of monoamine oxidase B was designed by means of computer moulding. This mould corresponds to the shape and volume size of selective substrates and inhibitors of this enzyme. Active site moulds of monoamine oxidases A and B exhibit significant differences in these parameters. The correctness of the resultant model of monoamine oxiadse B active site was validated by using three compounds which were not employed in the training sets. Weak inhibitors were not accommodated in the mould whereas their "flexible" analogues exhibitng rather potent inhibition are readily accommodated in this mould. This suggests that our mould reflects some important spatial features of monoamine oxidase B active site. This mould can be employed for screening of new perspective inhibitior of monoamine oxidase B and pilot evaluation of their monoamine oxidase B inhibitory activity. PMID- 11925757 TI - ELISA test to detect Chlamydophila pneumoniae IgG. AB - A new ELISA test (Chlamydophila pneumoniae IgG, Vircell, Spain) to detect Chlamydophila pneumoniae IgG was evaluated. The micro-immunofluorescence (MIF) test was used as reference method. Chlamydia trachomatis and Chlamydophila psittaci elementary bodies were also assayed. Two hundred and sixteen sera were included in the study: 66 from patients with peripheral arterial occlusive disease (Panel 1), 68 from adults with pneumonia (Panel 2), 44 from healthy adults (Panel 3) and 38 from patients with a sexuality transmitted disease by C. trachomatis (Panel 4). In Panel 1, 51 sera (77%) had antibody titres between 32 and 128; 4 out of 15 sera with IgG titres < 32 were positive by ELISA test and 2 sera with 32 IgG titres were uncertain by ELISA; the remaining 60 sera were correctly classified, giving a 91% concordance between the techniques. In Panel 2, 55 sera (81%) had IgG titres between 32 and 512; 2 out of 13 sera with IgG titres < 32 were positive by ELISA and 2 sera with 32 titres were uncertain by ELISA; the remaining 64 sera were correctly classified, giving a 97% concordance. In Panel 3, 22 sera (50%) had IgG titres between 32 and 64; only 1 out of 22 sera with IgG titres < 32 was positive by ELISA, giving a 97% concordance between the techniques. In Panel 4, there were 24 (63%) negative, 10 (26%) uncertain and 4 (10%) positive results by ELISA, giving an 86% concordance. The C. pneumoniae ELISA test demonstrated 100% sensitivity and 85% specificity. The IgG ELISA test demonstrated a good concordance with the MIF test without the drawbacks associated with the latter assay. We conclude that the ELISA test could be an alternative to the MIF test. PMID- 11925759 TI - Dependence of the sphingoid bases concentration on growth phase and temperature in the yeast Yarrowia lipolytica. AB - The aim of this research was to investigate the effect of growth phase and temperature on the concentrations of the individual molecular species of sphingoid based obtained by acid hydrolysis of total sphingolipids from the yeast Yarrowia lipolytica. Our results showed C18 phytosphingosine to be the major long chain base in Y. lipolytica regardless of growth phase or temperature. We also found Y. lipolytica to contain sphingosine, the predominant mammalian long-chain base that is uncommon for yeast sphingolipids. Among the identified long-chain bases, only C18 phytosphingosine appeared to be influenced by culture conditions. Its concentration was largest in the exponential phase and decreased 2.9-fold when cells entered the stationary phase of growth at 28 degrees C. Following a temperature shift from 28 to 39 degrees C, there was a 2.1-fold decrease in the phytosphingosine concentration, but it rose 1.7-fold after the heat-stressed cells had been returned to 28 degrees C and subjected to prolonged growth. These results might point to the possible involvement of phytosphingosine in the cell growth regulation and in the adaptation of Y. lipolytica cells to stressful culture conditions. PMID- 11925758 TI - Aerobic co-metabolism of sulfur, nitrogen and oxygen heterocycles by three marine bacterial consortia. AB - Bacterial samples were collected from three marine beaches in coastal Newfoundland, Canada, and enriched by growth on 1-methylnaphthalene. The most prominent bacterial cell type for each consortium was isolated in a serial dilutions test, and a substrate utilization profile was obtained for each using the Biolog MicroStation System. Each bacterial community was tested for its ability to co-metabolize sulfur heterocycles (benzothiophene: BT, 3 methylbenzothiophene: 3-MBT, and dibenzothiphene: DBT), a nitrogen heterocycle (carbazole: CARB), and an oxygen heterocycle (dibenzofuran: DBF). Co-metabolism of the starting material was determined using gas chromatography-mass spectroscopy (GC-MS), and formation of products was investigated by GC-MS and Fourier transform infrared (FTIR) spectroscopy. Bacterial growth was monitored turbidimetrically to determine the dry weight (microgram) of cells/ml. The 2 ringed heterocycles were co-metabolized faster and to a greater extent than the 3 ringed compounds. Co-metabolism of BT was not statistically different from that for 3-MBT and, likewise, a comparison of the 3-ringed heterocycles showed no significant differences in degradation rates. Statistical examination showed that no one culture demonstrated a significantly greater ability to co-metabolize the heterocycles studied. This study represents the first comprehensive investigation of the ability of local bacteria to co-metabolize a range of aromatic compounds and provides a preliminary understanding of their fate in sediments should contamination by these compounds occur. PMID- 11925760 TI - Alternative method of inoculum and spawn production for the cultivation of the edible brazilian mushroom Pleurotus ostreatoroseus SING. AB - Efficiency of solid and liquid inocula and their use for spawn production were compared so that improved cultivation conditions for the edible mushroom Pleurotus ostreatoroseus could be tested. Solid and liquid inocula were prepared respectively with Potato Dextrose Agar (PDA) and Liquid Potato Dextrose (LPD). Wheat grains and cotton residues were used as substrates for spawn preparation. Inoculum types did not affect the development of P. ostreatoroseus, and LPD spawns were cheaper, more homogenous, less contaminated. Decomposition activity of mushroom growth, as a percentage of organic matter loss (OML), was higher in the wheat grain spawn and was not influenced by the inoculum type. Advantages in the use of cotton residue for spawn production were longer storage time, lower contamination and reduced costs. The cotton residue substrate may be also used for the production of mushroom fruiting bodies. PMID- 11925761 TI - Fingerprinting of Nigerian Helicobacter pylori isolates by plasmid profile and PCR. AB - Plasmid profiling and digestion of amplified PCR product of ureA genes were used to determine genomic variation in 56 strains of Helicobacter pylori isolated from patients with peptic ulcers and subjects with gastritis recruited in Lagos and Ife, Nigeria. Twenty-five (45%) of the strains were found to harbour plasmids ranging in size from 0.9 kb to > 10 kb. The plasmid profile was able to detect differences between the strains, and also to distinguish between different strains isolated from the same patient. The expected amplified ureA gene PCR product was detected in all strains and digestion with the restriction enzyme DdeI did not produce discrimination amongst the strains, however, digestion with MluI produced little discrimination amongst strains. In conclusion, plasmid profiling produced better discrimination amongst H. pylori strains than ureA PCR gene profiling. PMID- 11925762 TI - Emulsifier from a tropical marine yeast, yarrowia lipolytica NCIM 3589. AB - A tropical marine strain of Yarrowia lipolytica, NCIM 3589 produced emulsifier in the presence of alkanes or crude oil. The mode of alkane uptake in this organism was by attachment to large droplets. An emulsifier (lipid-carbohydrate-protein) complex was associated with the cell wall. This emulsifier increased the hydrophobicity of the cells during the growth phase. In the stationary phase, the organism produced the emulsifier extracellularly under conditions of carbon excess and nitrogen limitation. Other requirements for extracellular emulsifier production included an initial pH of 8.0 and the presence of sodium chloride at a concentration of 2 to 3% (342 to 513 mM). The cell-associated and extracellular emulsifier was shown to have similar properties. PMID- 11925763 TI - Regulation of pyrimidine synthesis in Pseudomonas mendocina. AB - Regulation of the de novo pyrimidine biosynthetic enzymes in the bacterium Pseudomonas mendocina was examined when its cells were grown on succinate as a carbon source. When P. mendocina was grown in the presence of orotic acid or uracil, the de novo enzyme activities were depressed with dihydroorotase activity being significantly depressed after uracil addition. Following pyrimidine limitation of a uracil auxotroph of P. mendocina deficient for orotate phosphoribosyltransferase activity, the pyrimidine biosynthetic pathway enzyme activities were affected indicating possible regulation at the level of enzyme synthesis. Of the de novo pathway enzymes assayed, dihydroorotate dehydrogenase exhibited the highest increase in its activity. The regulation of the pyrimidine biosynthetic pathway by pyrimidines in P. mendocina appeared similar to what was previously observed for the taxonomically-related species P. stutzeri. PMID- 11925764 TI - [Objective reporting--how do we do that in MMP?]. PMID- 11925765 TI - [Bosentan in pulmonary hypertension]. PMID- 11925766 TI - [Progressive muscle relaxation according to E. Jacobson. A modern relaxation technique]. PMID- 11925767 TI - [Manual lymph drainage--indications, uses and risks]. PMID- 11925768 TI - [Vasoprotection with vascular nitric oxide. The effect of physical training and drugs]. PMID- 11925769 TI - [Psychosocial therapy of patients with dementia]. AB - The progressive cognitive and behavourial disturbancies in dementia diseases cause great emotional burden to patients, their families and caregivers. To relief this burden and to facilitate adequate coping strategies, psychosocial therapies intend to improve cognitive functions, self care abilities and emotional wellbeing of dementia patients. The therapeutic interventions address the patients, their caregivers and the adaption of the environment. Psychosocial therapies can be divided into cognitive training methods, behaviour orientated concepts, emotional orientated approaches and family interventions (1, 2, 12, 19). Most of these psychosocial therapies are only poor validated in randomized controlled trials. Nevertheless, they hold great promise to improve the quality of life and wellbeing of dementia patients and their caregivers. Thus they should be considered to be an integral part of a comprehensive therapeutic approach to dementia patients and their families. PMID- 11925770 TI - [Family caregiver help and self-help in Alzheimer dementia]. AB - Beside the loss of the memory capacity, non-cognitive disturbances occur up to 70%-90% in patients suffering from Alzheimer's disease due to pathological changes in the brain. Delusion, hallucination and changes of the circadian rhythm can appear in addition to the five kinds of disorder--agitation, aggressive behaviour, screaming, depression and constant hyperkinesia. The consequences of these changes in perception and behaviour constitute severe problems for the patient as well as for the main caregiver. The burden of caring often exceeds their energy and resources. Not only do many of those caregivers suffer themselves from exhaustion but also from feelings of guilt and depression. The therapeutic concept includes the involvement of the relatives through information, support, counselling and guidance as much as the investigation of the causes and interrelation of the problematic behaviour in each individual case and further involves the carer in creating a concept to deal in an optimal way with the patient. PMID- 11925771 TI - [Epidemiology of dementia]. AB - Because of the increasing life expectancy during the next decades the number of demented will steeply rise. The estimated number of people suffering from dementia in Austria will increase from 90,500 persons in the year 2000 to 233,800 persons in the year 2050. During the same period the estimated number of those developing a dementia within one year will rise from 23,600 to 59,500. In contrast, in the next five decades, the population being in an employable age will decrease to 77%. Thus, the proportion of demented in relation to employable people will considerably increase. While in year 2000 there were 56 employable persons per one demented person, in the year 2050 there will be only 17 employable persons per one demented person. Dementing disorders are a main reason for being admitted to nursing homes. In Austrian nursing homes 63.5% of those newly admitted suffer from dementia. Despite of the fact that a large proportion of dementia sufferers are being cared in nursing homes, the majority lives in their private households supported by their family members. Relatives of demented persons experience a large amount of burden, and need more professional help than is available today. The costs of dementing disorders are high and a large proportion is paid by the patients family. In the future, in Austria more specialized day care facilities for the demented and more nursing homes are necessary. PMID- 11925772 TI - [Differential diagnosis of dementia]. AB - Owing to the progress of individual life expectancy, the number of demented people will clearly increase in the next decades. Cognitive impairment refers not only to dementia, but also to mild cognitive impairment (MCI), associated with reduced complex daily activity and decreased quality of live. It may, in many cases (10-15% per year) progress to permanent dementia. The most frequent aetiologies of dementia syndromes are the Alzheimer's Disease (AD), vascular Dementia (VD), the combination of both and Lewy-Body-Disease. Primarily neuro psychiatric and internal medical reasons for cognitive decline have also to be taken into account for rational treatment. PMID- 11925773 TI - [Depressive pseudodementia]. AB - Elderly depressive patients complaining about cognitive symptoms are at particular risk of being labelled as demented. It is well documented that depressive disorders frequently cause mild cognitive deficits which manifest in psychometric procedures. A wide spectrum of potentially reversible cognitive deficits related to a depressive syndrome are summarized under the term of "Depressive Pseudodementia (DPD)". Most depressive patients who are referred to "DPD" suffer from cognitive dysfunctions outside the range of dementia. The clinical interface between depression and dementia is complex. There is some evidence that depression may be a risk factor for the expression of Alzheimer's disease in later life and that depression may occur as a prodrome for Alzheimer dementia. Moreover, depression often complicates the course of dementing disorders. However, there is no evidence that depressive disorders cause dementia without coexisting depressive symptoms. It is essential to search for depressive symptoms even after cognitive symptoms have been found. PMID- 11925774 TI - [Early diagnosis of dementia]. AB - Therapeutic interventions for dementia are crucially dependent on the early diagnosis of the case. The question is, how an early diagnosis on the symptom-, the syndrome- and the disease level can be accomplished. The data from epidemiological and clinical studies are described. On the symptom level a disorder of episodic memory, language and visuo-spatial functions have to be assessed in contrast to normal ageing effects and premorbid low cognitive functioning. Simple, clinically practical tests are described like an 10-word learning-test with encoding fascilitation, the semantic fluency (animals) and the clock drawing test. At the syndrome level the dementia syndrome of Alzheimer type has to be differentiated from other dementia syndromes, the senile amnestic syndrome and the delirium syndrome. At the disease level, the attempt to identify clinically the etiological entity is at most not possible with reasonable reliability in senile dementia cases. An improvement of the early diagnosis by prospective longitudinal investigations of the cases seems to be possible in the future. PMID- 11925775 TI - [Etiology and pathogenesis of Alzheimer dementia]. AB - Alzheimer's disease (AD) is the most common cause of primary dementia, characterized by a progressive process of pathophysiological restructuring of the brain over decades. The hallmark of Alzheimer's disease is the extracellular accumulation and deposition of insoluble amyloid, to be found in the parenchyma in the form of amyloid plaques and in meningeal and cerebral vessels as a congophile angiopathy. Equally conspicuous is the intraneuronal occurrence of neurofibrillary tangles, consisting mainly of hyperphosphorylated tau-protein. Amyloid plaques and neurofibrillary tangles are characteristic, but not specific to Alzheimer's disease. Similar changes can be found in healthy ageing processes and in various other neurodegenerative diseases. It is common to differentiate between an early-onset, familial Alzheimer's disease with an established genetic etiology, representing only about 5% of all cases, and the more typical late onset, sporadic Alzheimer's disease with an age of onset above 65 years and no clear pattern of inheritance. Although there seems to be a large heterogeneity in the etiology of Alzheimer's disease, the amyloid-cascade-hypothesis has taken a central position as a model for the general etiopathogenesis. The regulation of amyloid plaques underlies a diversity of cellular and molecular factors. In addition to ageing, apolipoprotein E 4 is a firmly established risk factor. Disturbance in the cerebral glucose metabolism, especially in the hippocampal regions, is a further proposed factor in the pathogenesis of Alzheimer's disease. The wide-spread loss of cortical cholinergic neurotransmission associated with the cognitive deficits is of importance to the comprehension of the symptoms and the present pharmacotherapy of Alzheimer's disease. PMID- 11925777 TI - [Dementia with Lewy bodies]. AB - Dementia with Lewy bodies (DLB) is the second most frequent neuropathologically diagnosed degenerative dementing illness. The clinical characteristics are progressive dementia, Parkinson syndrome, fluctuations of cognitive functions, vigilance and attention, visual hallucinations (usually detailed and well described), depression, REM-sleep behavior disorder, adverse responses to standard doses of neuroleptics, falls, syncopes, systematized delusions, and non visual hallucinations. Mean age at disease onset ranges between 60 and 68 years. Male persons are more frequently affected than female. Disease duration is six to seven years. The differential diagnoses of DLB are dementia of the Alzheimer type, Parkinson's disease, subcortical arteriosclerotic encephalopathy, progressive supranuclear palsy, multiple system atrophy, and, in rare cases, Creutzfeldt-Jakob disease. The genetic background of the disease is unclear. Magnetic resonance imaging and single photon emission tomography can contribute to the diagnosis. The disease is treated with L-dopa, atypical neuroleptics, acetylcholine esterase inhibitors, antihypotensive agents, and peripheral anticholinergic and alpha-receptor-blocking medicaments to improve neurogenic bladder dysfunction. PMID- 11925776 TI - [Symptomatic therapy of Alzheimer dementia]. AB - No causal treatment or highly effective preventive strategies are available for Alzheimer's disease to date. Symptomatic measures are still essential to improve cognitive and behavioural deficits. Acetylcholinesterase inhibitors, the NMDA antagonist memantine and the phytopharmaco ginkgo biloba are currently the substances most extensively studied and most widely used to improve cognition and activities of daily living. Behavioural disturbances oftentimes respond to antidementia drugs, but may demand specific treatment with neuroleptics, antidepressants, stabilizers and other substances. PMID- 11925778 TI - [Vascular dementia]. AB - The past decade has seen a renewed interest in vascular dementia. The search for a causal relationship between a vascular event or a vascular cerebral lesion and dementia has led to new classification schemes which no longer consider vascular dementia a homogeneous entity but acknowledge the diversity of the clinical and morphological substrates of this syndrome. Deviation from the term "multi-infarct dementia" is only one but many consequencies of these recent developments. Etiologically, vascular dementia may result from cerebral small vessel disease leading to extensive leucencephalopathy or lacunes or may be the consequence of strategically located infarcts or multiple infarcts in large vessel territories. It may also be the consequence of global cerebral hypoperfusion, intracerebral hemorrhage or other mechanisms such as vasculitis. There is no definitive medical or surgical treatment for vascular dementia. Thus, it appears that stroke prevention offers the most immediate and substantial solution to reduce the morbidity and mortality. This is best substantiated for treatment of arterial hypertension. Once vascular dementia occurs control of vascular risk factors may be useful but this contention will require larger scale studies to provide more definite proof. A number of metabolically active drugs has been used for the treatment of cognitive symptoms in vascular dementia. Yet, the data are conflicting und the effects described modest at most. There is epidemiological evidence for a more than incidental co-existence between vascular and primary degenerative dementia which suggests that therapies found to be effective in Alzheimer's disease may also prove beneficial at least in subgroups of vascular dementia. Lately, this concept is tested by several studies on the efficacy of acetylcholinesterase inhibitors in vascular dementia. PMID- 11925779 TI - [Frontotemporal dementia]. AB - Frontotemporal dementia is a rare form of progressive intellectual deterioration. Its most prominent clinical features are alterations in personality, motivation, and social conduct whereas memory and orientation remain largely unimpaired. Several underlying neurodegenerative processes may be distinguished which are confined to the cerebral cortex in most cases but occasionally involve the basal ganglia and rarely the anterior horn cells. Most frequently, histopathological examination reveals a non-specific loss of neurons accompanied by reactive gliosis. In a minority of cases, globose intraneuronal inclusions and achromatic ballooned neurons are seen. These peculiar morphological changes are called "Pick bodies" and "Pick cells" after the neurologist Arnold Pick who worked in Prague. It can be difficult to identify frontotemporal dementia because its major symptoms mimicnon-organic psychiatric disorders including mania, obsessive compulsive disorder, schizophrenia, depression or personality disorder. Another problem of diagnosis is that all clinical instruments that are available for assessing cognition, activities of daily living, and non-cognitive symptoms have been tailored to the prototypic dementia in Alzheimer's disease and are less sensitive to the psychopathology of frontal lobe diseases. The burden that frontotemporal dementia imposes on caregivers is also completely different from the one encountered by families of patients with the more prevalent dementias. In the present contribution we summarize the current evidence on epidemiology, aetiology, and risk factors of frontotemporal dementia. Clinical symptoms and course will be illustrated by a case example. We will also provide guidelines for diagnosis and discuss treatment options. PMID- 11925780 TI - [Alcohol-induced cognitive disorder: alcohol dementia]. AB - Cognitive impairment is frequently observed in patients with alcohol misuse or alcohol addiction. Multiple cognitive functions are reduced in these patients. Frontal lobe functions, as planning, abstract thinking, set shifting or continuous performance are most frequently affected. Alcohol amnestic syndrome, alcohol dementia and the Wernicke-Korsakow-Syndrome constitute distinct entities. Alcohol dementia follows the diagnostic criteria of dementia with clear evidence for alcohol abuse or alcohol addiction. The diagnostic procedure of alcohol induced cognitive impairment includes medical history, physical and neuropsychiatric examinations; laboratory examinations, neuropsychological assessment, brain imaging and electroencephalographic recordings. At the moment, there are no established treatment options for alcohol-induced cognitive impairment. Some evidence is provided that nootropics might be of benefit. Alcohol abstinence is a most important step. Psychosocial interventions are essential to support the patients in their daily activities. PMID- 11925781 TI - High resolution spiral computed tomography of the pancreas. AB - Computed tomography (CT) is a very useful tool in the assessment of pancreatic disease. Searching for subtle signs, as in chronic pancreatitis or staging of adenocarcinoma, high spatial and contrast resolution is needed. The high resolution computed tomography (HRCT) technique for pancreatic scans, and its evolution from dynamic CT to multislice spiral CT, is described. 2D and 3D dimensional reconstructions are depicted and their role in diagnosis is focused. Together with spatial resolution, contrast enhancement protocols are discussed, aimed to achieve optimal contrast between the lesion and normal parenchyma. PMID- 11925782 TI - Magnetic resonance imaging of the exocrine pancreas. AB - The currently available MRI, techniques that are used to perform imaging of the pancreas are reviewed. A basic pancreas examination includes: fat-suppressed breath-hold T1-weighted and respiratory triggered or breath-hold T2-weighted fast sequences The use of dynamic MRI with both extracellular gadolinium chelates and specific contrast agents is stressed in the evaluation of exocrine pancreatic diseases. It is concluded that an "all-in-one" approach including MR angiography of the pancreatic vessels and MR cholangiopancreatography of the biliopancreatic ductal system, confirmed by further studies, is presumably the most cost effective imaging technique in the evaluation of a vast array of exocrine pancreatic diseases. PMID- 11925783 TI - Magnetic resonance pancreatography (MRP): morphology and function. AB - MRCP is able to non-invasively assess the pancreatic ducts, with moderate/high agreement with ERCP: There is however a high number of false negative results, mainly due to the small size of the main pancreatic duct, especially in the tail, and side branches. Secretin stimulates the exocrine pancreas with accumulation of fluid and bicarbonates in the ductal system, and subsequent enlargement. This increase in caliber improves the assessment of the morphology of pancreatic ducts and their abnormalities. MRCP accuracy in assessing ductal abnormalities, improves after secretin administration. Furthermore, dynamic MRCP during secretin administration is also able to afford the functional evaluation of the pancreatic flow dynamics. Papillary stenosis, either idiopathic or due to Santorinicele, is easily diagnosed by means of S-MRCP. Furthermore a noninvasive assessment of the pancreatic exocrine reserve can be performed with dynamic MRCP during secretin administration. PMID- 11925784 TI - Imaging of acute pancreatitis. AB - Pancreatitis is one of the most complex and clinically challenging of all abdominal disorders. It is classified according to clinical, morphologic and histologic criteria. The primary role of radiologic imaging in patients with suspected pancreatitis is to confirm or exclude the clinical diagnosis of pancreatitis. Second, if possible, the cause of the disease is established with the assessment of disease severity and detection of complications. Imaging can also provide guidance for percutaneous therapy. Sonography in acute pancreatitis is a good screening test in patients with suspected biliary pancreatitis and a mild clinical course. Contrast-enhanced CT is preferred for patients with acute pancreatitis because it can accurately diagnose and stage the disease and the necessary information for percutaneous management is provided. The diagnosis of acute pancreatitis on MRI relies on the presence of morphologic and peripancreatic changes. Pancreatic necrosis and complications of acute pancreatitis such as hemorrhage, pseudocysts or abscesses are well-examined by MRI. PMID- 11925785 TI - Imaging of chronic pancreatitis. AB - Chronic pancreatitis is characterized by progressive, irreversible morphologic changes whose most common cause is excessive alcohol intake. Radiologic imaging plays a major role in the diagnosis, staging of disease severity, detection of complications and selection of treatment options. The sensitivity of US ranges from 60% to 70% while its specificity is higher reaching 80%-90% in the detection of abnormalities of main pancreatic duct. As for CT, its sensitivity and specificity in the diagnosis of chronic pancreatitis, in recent studies is 74% and 85% respectively. The performance of Magnetic Resonance cholangiopancreatography was shown to be enhanced by secretin stimulation with better visualization of ductal and parenchymal changes. However, endoscopic retrograde cholangiopancreatography is the most sensitive indicator of the presence and extent of the disease. Ductal abnormalities can be used to classify chronic pancreatitis. PMID- 11925786 TI - Staging of pancreatic ductal adenocarcinoma with spiral CT and MRI. AB - Detection of pancreatic adenocarcinoma is crucial for accurate staging both with spiral CT and dynamic MRI; consequently an accurate technique is required and so called pancreatic phase is recognized as the best one for tumor conspicuity. For vascular involvement optimal results have been achieved in assessing unresectability; vein involvement seems more difficult to be defined as it is sustained by a different spread compared to arterial involvement. Grading of vessel circumference contact represents the best tool in "venous" staging, but shape deformation and collateral veins dilation are also important signs. Lymph node staging is less accurate, lacking in specificity, but spiral CT demonstrated better results if compared with dynamic MRI. Assessment of liver metastases has been improved by the advent of spiral CT and dynamic contrast enhanced MRI, while peritoneal staging seems to be unaffected. In conclusion, both spiral CT and dynamic contrast enhanced MRI are accurate in pancreatic adenocarcinoma staging, mainly for vessels and liver involvement; no definite differences have been established, because only a few studies have compared them both with state-of-art techniques. Therefore standardized multicentric trials are desirable. Up to now, the choice of which technique to employ should be based on local expertise; moreover, the aggressive approach of surgical equipes should be kept in mind. PMID- 11925788 TI - Practical aspects in drawing up a quality handbook in radiotherapy. AB - The quality and dimensions of the Quality Handbook in healthcare are shortly defined. Particular emphasis is put on the role of the quality of compliance and on certification (ISO 9000). The role of quality assessment in healthcare processes is described. It is not conceived as an inspection but rather as an incentive to a continuous quality improvement in healthcare processes. Within the quality of compliance, the role of the Quality Handbook and its modalities of definition, organization and implementation are illustrated. PMID- 11925787 TI - Diagnosis of exocrine diseases of the pancreas: is there still a role for endoscopic retrograde cholangiopancreatography (ERCP)? AB - In the diagnosis of exocrine diseases of the pancreas, three major questions should be addressed: when only MR cholangiopancreatography should be applied; when endoscopic retrograde cholangiopancreatography should be applied directly; when a combined approach should be applied. In pancreatic disease MRI has many indications because ultrasonography, the procedure of choice, is rarely able to resolve the diagnostic problem, with a clearcut indication for therapeutic endoscopic retrograde cholangiopancreatography. One of the rare cases where MRI could be omitted is represented by pancreatic cancer with biliary obstruction in non operable patients in whom a pancreatic mass was already visualized on ultrasonography. In all other diseases of the pancreas, the combination of MRI/MRCP with secretin stimulation and ERCP usually favors the first procedure, preventing a number of diagnostic ERCP. PMID- 11925789 TI - Continuous quality improvement in radiotherapy: the role of the "Istituto Superiore di Sanita" in its application over the national territory. AB - The "Laboratorio di Fisica" of the "Istituto Superiore di Sanita" has undertaken some initiatives to tackle the problem of patients' radiation protection in relation to the issues of radiotherapy, both with respect to the promulgation of new National and European laws and directives and to the extremely rapid and innovative technological developments involved in this field. First, interdisciplinary study groups were organized to identify the aspects on which recommendations to be applied at the national level could be based, and to provide for their development. Second, a national plan was launched for continuous quality improvement in radiotherapy, intended as a program of controls and correctives implemented in radiotherapy to ensure its compliance with predefined standards together with its continuous improvement. Within this plan, a group of experts coordinated by the "Istituto Superiore di Sanita" has promoted two different intercomparisons of dosimetry to evaluate the differences between the measured dose and the calculated dose 1) with reference to a water phantom and 2) during a simulated treatment of rectal cancer in a plastic phantom. PMID- 11925790 TI - Radiotherapy quality assurance: international and national recommendations. PMID- 11925791 TI - Regulatory aspects of health service accreditation. AB - The process of accreditation is analyzed in light of the national "Atto di Indirizzo e Coordinamento per l'Accreditamento Istituzionale". While assessment criteria are established at the national level, the determination of requirements is assigned to the Regions. PMID- 11925792 TI - Radiation oncology practice accreditation. AB - The practice accreditation program of The American College of Radiology is described. Practice accreditation constitutes a facile mechanism to accomplish external quality assurance and to assess compliance with recognized standards. The process of accreditation includes critical review of each radiation therapy facility, the technical staff, all quality assurance procedures, medical physics staff, nursing and physician personnel as well as a whole host of other items, the most important of which is meticulous examination of selected patient charts. In the appendix, standards developed by the American College of Radiation Oncology, are reported. PMID- 11925793 TI - Continuous quality improvement in radiotherapy: implementation methodology and comparison of experiences. AB - The Quality System (QS) was applied in the Italian Health Service in the late nineties with the identification of a methodology whose focus is also on nonspecific subjects which however are involved in health promotion. The quality handbook represents an instrument of QS, where the documentation and procedures which can make the policy and goals of a center accurately and extensively known, are collected. Continuous quality improvement is characterized by the adoption of the assessment culture according to dynamic processes. Since several years, in Italy, documents indicating the requirements, resources and goals radiotherapy centers should aim at, are being prepared together with the identification of means suitable for quality assurance in radiotherapy. PMID- 11925794 TI - Quality handbook in radiotherapy: personal experience. AB - The Quality Handbook includes the description of the activities carried out in Radiotherapy and the methodology used to ensure the Quality Assurance according to the principles indicated in national and international documents of reference and the recommendations of accreditation agencies. The structure of the Quality Handbook, the main aspects of single chapters, the selected quality indicators undergoing inspections are analysed and procedures to be followed for changes and updating of the Quality Handbook are described. The Quality Assurance program is illustrated in another article of this issue. PMID- 11925795 TI - Quality assurance in radiotherapy: personal experience. AB - The approach to the implementation of a Quality Assurance Program applied from 1 October 2000 at the Radiotherapy Service of the "Universita Cattolica del S. Cuore, Policlinico "A. Gemelli" of Rome, is described. Some major aspects of this program are analyzed. PMID- 11925797 TI - [Decreased fetal movement and a sinusoidal pattern on the cardiotocogram: 2 alarm signals]. AB - A 24-year-old primigravida was referred at 35 weeks of gestation due to decreased foetal movement. External monitoring demonstrated a sinusoidal foetal heart rate pattern accompanied by late decelerations. A hydropic and severely anaemic infant with clinical signs of thrombocytopenia was delivered by emergency caesarean section. He failed to breath spontaneously and required positive pressure ventilation. Cranial ultrasound and computerised tomography of the brain revealed a massive intraparenchymal haemorrhage. After the infant had died, post mortem studies revealed the presence of maternal antibodies to the human platelet antigen (HPA) 1a which suggested neonatal isoimmune thrombocytopenia as cause of the intracranial haemorrhage. A sinusoidal foetal heart rate pattern is an alarm signal--it is not necessarily significant but is often associated with mortality or serious morbidity and is therefore a reason for quickly investigating the condition of the foetus. PMID- 11925796 TI - Quality handbook in radiotherapy. Brachytherapy: personal experience. AB - Brachytherapy is a conventional method of radiation therapy characterized by peculiar technical, clinical, operational and radioprotection problems. Therefore, the management of a service or department of brachytherapy requires a specific organization aimed at Quality Assurance. In this report, the personal experience with the drawing up of a Quality Handbook of brachytherapy with reference to the method used and the structure of the document, is described. PMID- 11925798 TI - [Arguments for offering triple test serum screening for Down's syndrome to all pregnant women]. AB - In an advisory report to the Dutch Government, the Health Council of the Netherlands has recently recommended offering triple test serum screening for Down's syndrome to all pregnant women. According to a critical review in this journal, the Council should have proposed excluding those less than 30 years of age. However, there is no age at which a logical limit can be set. Moreover, to suggest that the negative aspects of the screening will prevail for those under a specific age, denies the women involved the right to make a judgment based on their own values and preferences. Secondly, although limited resources may be a reason for limiting access to screening, doing so would be a politico-ethical decision, which raises the issue of justice. Finally, there are reasons for thinking that a new age limit would complicate the necessary counselling as opposed to making it easier. PMID- 11925799 TI - [The concomitant use of radiotherapy and chemotherapy for the treatment of solid malignant tumors]. AB - Various clinical trials carried out during the last few decades have clearly demonstrated that the concomitant use of radiotherapy and chemotherapy significantly improves local control in a variety of advanced solid tumours. In most of these trials where an improvement was noted, cisplatin was used in addition to radiotherapy either on its own or in combination with other cytostatics. This has led to improved survival rates in head and neck, lung and cervical cancer. For these solid tumours, the interaction of radiotherapy with chemotherapy appears to be schedule dependent, as no such an improvement was observed if chemotherapy was given prior to or after radiotherapy. A major advantage of this combined therapy is that a structure-preserving treatment for patients with advanced larynx or anal cancer is more often possible. Major further improvement can be expected from the design and use of new drugs that influence the pathways leading to cell death after irradiation. PMID- 11925800 TI - [Physical diagnosis--digital rectal examination]. AB - For a wide range of complaints, there is an indication for digital rectal examination. The position patients should adopt depends on their physical condition and the indication concerned. The reach of the palpating index finger is fairly short. The severity of micturition complaints has little or no relation to the size of the prostate. The sensitivity of digital rectal examination for detecting prostate carcinoma ranges from about 50 to 80%. Therefore, a prostate carcinoma cannot be excluded on the basis of digital rectal examination. The positive predictive value of digital rectal examination for detecting prostate carcinoma increases as the serum PSA level increases. Digital rectal examination can make an important contribution to the diagnosis of anorectal disorders, including rectal carcinoma. In total, 5-10% of consultations with general practitioners are related to anorectal or urogenital complaints and 50% of the elderly have micturition problems; therefore digital rectal examination is one of the physician's basic skills. PMID- 11925801 TI - [Diagnostic image (80). A men with thirst and polyuria. Bladder retention]. AB - A 69-year-old man was admitted because of polyuria and renal insufficiency. At physical examination a huge bladder retention was found (31). The polyuria was explained by the decreased concentrating ability of the kidney due to impaired tubular function; this was caused by hydronephrosis due to prostate hypertrophy. PMID- 11925802 TI - [Recurrent otitis media and mastoiditis due to atypical mycobacteria]. AB - An eleven-year-old girl was operated on due to right-sided chronic otitis media with effusion. After three months, an impressive enlargement of the mucosal lining developed, for which thorough debridement of the middle ear and mastoid was performed. Histological examination revealed a granulomatous inflammation, with negative Ziehl-Neelsen staining. Standard bacteriological cultures revealed no pathogenic micro-organisms. Three weeks later the same clinical picture developed, once again followed by extensive surgical debridement. After a thorough diagnostic work-up an atypical mycobacterium was found, namely Mycobacterium abscessus--formerly named M. chelonei subspecies abscessus. Following appropriate antibiotic therapy the patient was symptom-free. Mycobacterial infections should be part of the differential diagnosis of persistent otorrhoea. PMID- 11925803 TI - [A men with a Salmonella dublin-infected aneurysm of the abdominal aorta]. AB - A 71-year-old male was diagnosed with a Salmonella dublin infection. He presented with abdominal pain with no diarrhoea, and sepsis, and was found to have an infected aneurysm of the infrarenal abdominal aorta. He was treated surgically with resection of the aneurysm and implantation of an extra-anatomic axillobifemoral bypass, followed by long-term antibiotic treatment. Nine months after the primary treatment, the patient died as a result of rupture of the aortic stump. S. dublin-infected aneurysm of the abdominal aorta is a rare condition with high mortality. Human S. dublin infections are associated with the consumption of unpasteurised dairy products from infected animals. PMID- 11925804 TI - [Information about cancer treatment and the media]. AB - A 'hopeful' development in cancer treatment (angiostatin) was reported in the Dutch press, which led to considerable disquiet amongst patients. There were no scientific articles that warranted this press publication. It has previously been stated that it is unethical for medical researchers to inform the press about treatments which have not been scrutinized by the medical community. The press on the other hand, should maintain a more responsible attitude toward medical claims and double-check medical information, even when it comes from renowned medical researchers. The main distinction between mainstream medicine and medical quackery is that the former is based on the results of controlled published research. When physicians propagate results in the popular press that have not been critically evaluated by the medical community, the distinction between quackery and mainstream medicine is undermined. PMID- 11925805 TI - [Acute pancreatitis, a protocol for diagnosis and treatment]. PMID- 11925806 TI - [Acute pancreatitis, a protocol for diagnosis and treatment]. PMID- 11925808 TI - [Acute pancreatitis, a protocol for diagnosis and treatment]. PMID- 11925807 TI - [Acute pancreatitis, a protocol for diagnosis and treatment]. PMID- 11925809 TI - [Reduced risk of complications associated with severe acute (necrotizing) pancreatitis by administration of antibiotics; results from a literature review]. PMID- 11925810 TI - [Reduced risk of complications associated with severe acute (necrotizing) pancreatitis by administration of antibiotics; results from a literature review]. PMID- 11925811 TI - [Reduced risk of complications associated with severe acute (necrotizing) pancreatitis by administration of antibiotics; results from a literature review]. PMID- 11925812 TI - [Reduced risk of complications associated with severe acute (necrotizing) pancreatitis by administration of antibiotics; results from a literature review]]. PMID- 11925813 TI - [Reduced risk of complications associated with severe acute (necrotizing) pancreatitis by administration of antibiotics; results from a literature review]. PMID- 11925814 TI - [Deep venous thrombosis as a complication of congenital absence of vena cava inferior]. PMID- 11925815 TI - [Deep venous thrombosis as a complication of congenital absence of vena cava inferior]. PMID- 11925816 TI - [Female patient with cutaneous anthrax in Belgium]. PMID- 11925817 TI - [Spondylodiscitis in 3 children; differential diagnosis and treatment]. PMID- 11925818 TI - [Irritable bowel syndrome: pathophysiology and (future) treatment options]. PMID- 11925819 TI - [Acetaminophen use by chronic alcohol abusers: a therapeutic dose may be too much for the liver]. PMID- 11925820 TI - [Scombroid poisoning after eating tuna fish]. PMID- 11925821 TI - [A family with malaria: from asymptomatic to life-threatening]. PMID- 11925822 TI - [Diagnostic image (72). An adult woman with coarsening of the face. Acromegaly]. PMID- 11925823 TI - Facial appearance in adults who had cleft lip and palate treated in childhood. AB - Sixty-eight adults (44 men and 24 women) with treated total cleft lip and palate were compared with a group of 66 adults (49 men and 17 women) without clefts, matched by sex and age. The outcome measures included a self-report questionnaire about body image, quality of life in general, health-related quality of life, somatisation, and depression. The group with clefts were also asked if they had further need of treatment. They reported significantly more dissatisfaction with their facial appearance (nose, lips, mouth, profile, and overall facial appearance) than the group without clefts. Satisfaction with facial appearance was significantly correlated with a better quality of life in both groups, and significantly correlated with a better health-related quality of life and a lower grade of somatisation in the group with clefts. Dissatisfaction with facial appearance was the most significant predictor of depression in both groups. A panel of four professionals from a cleft lip and palate treatment team judged the outcome of treatment in 64 of the subjects on colour slides. The professionals and the subjects with clefts were generally not very satisfied with the results of the surgical outcome. Thirty of the subjects with clefts (47%) wished to have more operations. The professional group recommended further operations in 38 of the cases (59%), in particular, rhinoplasties. We conclude that the subjects with treated clefts were not satisfied with their facial appearance, although they seem to be psychosocially well-adjusted to their disability. PMID- 11925825 TI - Self-concept and introversion in adolescents with cleft lip and palate. AB - The purpose of this study was to investigate if adolescents with cleft lip, or palate, or both (CLP), have an altered self-concept, and to assess their degree of introversion, compared with a control group. The CLP group consisted of 55 adolescents (aged 17-20 years) and the control group consisted of 31 adolescents (16-19 years). The Tennessee Self Concept Scale (TSCS) was used to measure the subjects' self-concept, while the Eysenck Personality Questionnaire Inventory (EPQ-I) was used to measure introversion. The results indicate that those with CLP have a normal or even a high self-concept, and no signs of introversion. PMID- 11925824 TI - Functional recovery in a tendon autograft used to bridge a peripheral nerve defect. AB - The functional recovery was examined after two different methods had been used to bridge an extended nerve gap. A 15 mm defect in the rat sciatic nerve was repaired using a tendon autograft--a new graft material--on one side and a freeze thawed muscle graft--a well established experimental material--on the other side. Evaluation after 12 weeks included measurements of tetanic force in the gastrocnemius muscle and computerised morphometry of the tibial nerve. The muscular tetanic force recovered to 26% and 21% of control muscles, respectively, but there were no significant differences between the two types of grafts in any of the measurements. The number of regenerated axons of the tibial nerve correlated with functional recovery as judged by muscular tetanic force in the gastrocnemius muscle. We conclude that the tendon autograft supports functional recovery, as judged by return of muscular tetanic force, to an extent comparable to that of the freeze-thawed muscle graft. PMID- 11925826 TI - V-Y advancement flap for the reconstruction of partial and full thickness defects of the upper lip. AB - We analysed the records of 25 patients who had their upper lips reconstructed after resection of a tumour. All the repairs were done with triangular cutaneous and musculocutaneous flaps with a skin island with V-Y advancement, and a subcutaneous or muscular pedicle with or without mucosa. The choice of flap was based on the width of the defect after the tumour had been resected. A subcutaneous pedicled flap was used in 14 patients; a musculocutaneous flap not including the mucosa in 5, and including the mucosa in 6. This flap can be used to repair upper lip defects of any thickness. The procedure is quite safe from a circulatory point of view and the flap has the advantage of requiring only one procedure. It is aesthetically satisfactory in most patients and maintains the good function and sensitivity of the lip. PMID- 11925827 TI - Reconstruction of the upper extremity with a compound rib-latissimus dorsi osteomusculocutaneous flap. AB - The reliability and versatility of the pedicled latissimus dorsi muscle or osteomusculocutaneous flap make it our first choice in the management of upper arm injuries and we have treated three such patients in this way. They had severe skeletonising, crushing injuries of an upper extremity with humeral defects that were treated with latissimus dorsi musculocutaneous flaps and segments taken from the ribs. All the flaps survived completely with no injury of the pleura at the donor site. The reconstructed humerus was strong enough for the patients to participate in all activities of daily living. We think that this technique is suitable for the upper arm defects with humeral loss because of its simplicity and minimal morbidity. PMID- 11925828 TI - Patient-rating of long term results of silicone implant arthroplasty of the scaphoid. AB - Thirty-two patients had silicone implant arthroplasty of the scaphoid in our unit between 1974-1988. Ten years later eight had had their implants removed because they had failed. Twenty-four of the 32 were reviewed at an intermediate clinical follow up examination between 1988-89 and 21 by patient rated wrist evaluation 10 years later. Grip strength compared to the non-injured hand varied between 38% and 136% (mean (SD) 78 (25)%). Nine of the 24 wrists showed no signs of bone cysts, while 15 had intraosseous cysts of various sizes. Patient-rated evaluation gave a median subjective long term overall wrist score of 25, a pain score of 27, and a function score of 22 on a scale from 0 (best) to 100 (worst). Nine out of 21 patients had little or no pain, and 13 out of 21 had total scores of under 25. Many of these patients reported good subjective benefit from silicone scaphoid arthroplasty. PMID- 11925829 TI - Granular cell tumour of the ulnar nerve in a young adult. AB - Granular cell tumours in peripheral nerve trunks are extremely rare. We report a case of a granular cell tumour of the ulnar nerve just distal to the elbow in a 16-year-old man. The appearance of the intraneural tumour could not be differentiated by magnetic resonance imaging (MRI) from more common nerve tumours. At exploration it was impossible to excise the tumour radically. Microscopic examination showed a clearly benign tumour and no malignant transformation has been found at follow-up after three and a half years. PMID- 11925830 TI - Skin expansion and external tissue extension techniques in the treatment of a traumatic scalp defect. AB - Large scalp defects that include periosteum set surgeons problems in closing the defect and in reconstruction of the hair-bearing area. The external tissue extension (ETE) technique together with a skin expansion device allowed us to reduce a 12 x 7.5 cm scalp defect and to construct a self-closing flap. PMID- 11925831 TI - An acquired giant vascular tumour of the breast. AB - We present a rare case of an acquired giant vascular tumour of the breast that was diagnosed as angiomatosis. It was characterised by the mixture of blood filled haemangiomatous, and apparently empty lymphangiomatous, vascular channels. The breast remnant returned to a normal configuration and contour without breast reduction. PMID- 11925832 TI - Primary leiomyosarcomas of the skin. AB - We report four cases of cutaneous, and one of subcutaneous, leiomyosarcoma with the results of histological examinations and immunohistochemical studies. Subcutaneous leiomyosarcomas have more likelihood of metastases than the cutaneous type. Only one of our cutaneous leiomyosarcomas recurred after 18 months and the patient with a subcutaneous lesion had lung metastases after five years. PMID- 11925833 TI - Rare case of a rudimentary medial metatarsal non-ossified structure. AB - A 9-year-old boy presented with a rudimentary medical metatarsal non-ossified structure. We considered his condition to be classified as hypoplastic medial member type in the metatarsal type of medial ray polydactyly. When it was considered as polydactyly, it had the longest delay of ossification among reported cases. PMID- 11925834 TI - Otitis media and feeding with breast milk of children with cleft palate. AB - The purpose of the present study was to analyse the incidence of acute and secretory otitis media (OM), and feeding with breast milk, and the use of a grommet in children with a cleft palate (CP/CLP) or cleft lip (CL), compared with controls. A total of 84 children between 6 and 10 years of age were studied. The CP/CLP group consisted of 48 children with an isolated cleft palate (n = 28), or a cleft lip and palate (n = 20). The CL group consisted of 15 children with an isolated cleft lip. The controls were 21 children without clefts. Children with CP/CLP had acute OM significantly more often than children without clefts (43/48 compared with 10/21), and secretory OM (40/48 compared with 4/21), despite the use of grommets. CP/CLP children were breast fed for a mean of 2.8 months (range 0-13), compared with 3.6 months (0-12) for CL, and 7.5 (0-24) months for controls. There was a significant correlation during the first 18 months of life between longer duration of feeding with breast milk and a lower incidence of acute and secretory OM in the three study groups combined. The incidence of otitis media was not affected by care in a day centre, having a sibling attending a day care centre, or by the family's medical history. Despite cleft repair and early treatment with grommets, both secretory and acute OM are common among children with cleft palate, presumably as a result of their eustachian tube dysfunction. The present study suggests that premature cessation of feeding with breast milk may contribute to an increased incidence of acute and secretory OM. PMID- 11925835 TI - The Dutch experience. AB - Euthanasia has been legally sanctioned in the Netherlands by a series of court decisions going back to the 1970s. The author discusses the cultural and historical factors that may have contributed to this development. In the past decade, studies sanctioned by the Dutch government reveal that guidelines established for the regulation of euthanasia--a voluntary, well-considered, persistent request, intolerable suffering that cannot be relieved, consultation with a colleague, and reporting of cases--are consistently violated. Of greatest concern is the number of patients who are put to death without their consent- there are more involuntary than voluntary cases. Euthanasia intended originally for the exceptional case has become an accepted way of dealing with the physical and mental distress of serious or terminal illness. In the process palliative care has become one of the casualties while hospice care lags behind that of other countries. Case examples are given. PMID- 11925836 TI - A critique of Callahan's utilitarian approach to resource allocation in health care. AB - The rationale of this article is grounded in the liberal tradition. It places the individual at the center of concern, and attempts to fortify the individual's basic right to health care. Attention is focused on the writings of Daniel Callahan, arguing that his approach is too cold and detached, and that age should not serve as the decisive criterion. The criticism of his views on older patients and on patients in post-coma unawareness (PCU) stems from two different lines of reasoning: the medical and the moral-contractual. From the medical perspective, while age is an important variable in determining a patient's medical condition, there are other--no less important--factors that influence one's health. From the moral-contractual line of reasoning, liberal society should not desert its citizens at the time they need its help most. The age criterion is too simple, too general, too sweeping. It provides too convenient an answer to a tough and troubling question. Similarly, the argument with regard to PCU patients should be qualified, taking into account the age of the patient, the cause of the condition, and the length of time in state of unawareness. PMID- 11925837 TI - Dispensing of controlled substances to assist suicide: a memorandum of the Office of the Attorney General, the United States Department of Justice. PMID- 11925839 TI - [Progress in the development of new generation antipsychotic agents and cognitive behavior therapy for patients with schizophrenia]. PMID- 11925838 TI - [Cognitive behavior therapy for schizophrenia with delusion and hallucination]. PMID- 11925840 TI - [Acute phase drug therapy and drug switching in the treatment of patients with schizophrenia]. PMID- 11925842 TI - [Drug therapy for patients with mood disorders]. PMID- 11925841 TI - [Cognitive therapy combined with drug therapy for the treatment of patients with depressive disorder]. PMID- 11925843 TI - [Comorbidity in depressive disorders]. PMID- 11925844 TI - [Therapy for patients with substance abuse from point of view of brain science and spirituality]. PMID- 11925845 TI - [Present condition and future prospects for repetitive transcranial magnetic stimulation therapy]. PMID- 11925846 TI - The external quality assessment schemes in Thailand. AB - In Thailand, the external quality assessment schemes have been organized by 2 main institutions:--The Department of Medical Science, Ministry of Public Health and The Faculty of Medical Technology, Mahidol University. Both schemes were initiated simultaneously by WHO experts in 1973. The Department of Medical Science, Ministry of Public Health established a Unit of Laboratory Quality Standards to be responsible for external quality control for clinical laboratory services. In 1996 the unit was upgraded to become The Bureau of Laboratory Quality Standards and is responsible for The National Proficiency Testing Scheme (NPTS). This includes hematology, clinical chemistry, clinical immunology, clinical microscopy, clinical microbiology and blood banking. Every laboratory is invited to be a member, free of charge. Now there are almost 800 out of 1300 laboratories all over Thailand participating in this NPTS. The proficiency testing samples are sent to the participants 3-4 times/year. The results of laboratory tests performed by participants are evaluated by using target values for every scheme, except clinical chemistry, which use participants' consensus. The control materials used in the clinical chemistry, hematology and immunology schemes are imported from aboard. The remaining control materials are prepared in house. The Faculty of Medical Technology, Mahidol University has organized 4 programs in The External Quality Assurance Scheme (EQAS):--clinical chemistry, clinical hormone, clinical microscopy and clinical immunology and serology. The first scheme was established in 1986 and the remaining schemes were established in 1999. All the control materials used are prepared in house. The members of this EQAS have to pay a membership fee. The control samples are sent to the participants 4-12 times/year. The results of laboratory tests are evaluated by using participants' consensus. There are 150-460 laboratories enrolled in this EQAS. At this time, Thailand is very conscious of quality in every field, including hospitals, and internal and external quality controls are one of the recommendations of the quality standard. So both The NPTS and The EQAS mentioned above are very important for each laboratory at the moment. The author sent questionnaires to 200 laboratories asking whether they were enrolled in an external quality assessment scheme. Fifty-seven laboratories responded and over 70% of them had joined either The NPTS or The EQAS and some of them had joined both. In addition, there are 2 new programs of external quality control:--the external quality control in red cell serology and the external quality assessment of hematology laboratory which have been established recently. However, there are still some types of laboratory that have no external quality assessment programs e.g. coagulation, serology for autoimmune disease and hemoglobin typing. External quality assessment programs for these laboratories are urgently needed. PMID- 11925847 TI - External quality assessment scheme and laboratory accreditation in Indonesia. AB - The National Program on External Quality Assessment Scheme (NEQAS) in Indonesia was first started in 1979, organized by the Indonesian Ministry of Health collaborating with professional bodies. The first trial was for clinical chemistry test with 2 cycles per year, followed by the hematology NEQAS in 1986 in collaboration with WHO-Royal Post Graduate Medical School London. After that, the schemes for serology, microbiology and parasitology were also organized. Around 500-600 laboratories throughout Indonesia participated each year in these quality control schemes, 2-4 cycles per year. Samples would be sent to participants and results will be given back to each laboratory. Poor performers should participate in the workshop or training course conducted by the Central Health Laboratory to improve their results. Participation in this NEQAS is mandatory for obtaining the laboratory license, and the Ministry of Health uses these schemes as one of the means for monitoring and coordinating the performance of laboratories throughout Indonesia. There are also some other EQAS (External Quality Assessment Scheme) programs conducted by professional bodies, such as for hemostasis, clinical chemistry and serology. During the course of conducting these schemes, it could be observed that manual methods were gradually changed to the automatic methods, especially for the clinical chemistry and hematology laboratories, which counts also for improvements of their results. Since the last 6 years, the Ministry of Health also began to conduct the Accreditation System evaluation for hospitals, including the laboratory departments. There are 7 standards that were evaluated, such as the aspect of the organization, administration and management, staffing, facilities and equipment, standard operating procedures, research and developments and quality control. This accreditation program is still in progress for all public and private hospital laboratories. PMID- 11925848 TI - External quality assurance schemes: a local perspective. AB - Medical laboratories in Singapore subscribe to External Quality Assurance Schemes run by the Licensing and Accreditation Unit of the Ministry of Health, Singapore, as well as a number of foreign medical laboratory quality assurance institutions. Through these organisations, a comprehensive range of laboratory tests are externally monitored for quality assurance at regular intervals in all medical laboratories. In recent years, an increasing number of local quality assurance schemes have been implemented, and an increasing number of laboratories have gained accreditation by foreign quality assurance institutions. The respective roles of these quality assurance activities are discussed. PMID- 11925849 TI - [Unexpected drug-interaction]. AB - The case of a male patient suffering from chronic normal pressure hydrocephalus is outlined. Antidepressant and pravastatin were administered because of the patient's abulia and hypercholesterolemia, but neuroleptic malignant syndrome like conditions developed. All physicians should suppose the occurrence of such an "unexpected drug-interaction" in any case. The author considered that a good sense of careful discernment and rapid reference system of medical information are "essential tools" for clinical management. PMID- 11925850 TI - [Relationship of polymorphism in CYP2C9 to genetic susceptibility to diclofenac induced influenza-virus-associated encephalopathy]. AB - The mechanism causing influenza-virus-associated encephalopathy is unclear, even though diclofenac metabolites may induce this pathogenesis. CYP2C9 is known as the major cytochrome P450 gene product that catalyzes diclofenac in human liver. It is uncertain whether the mutation of CYP2C9 is associated the pharmacologic effects of diclofenac in influenza infection. Therefore, we applied a simple and rapid procedure involving real-time fluorescence allele-specific PCR(TaqMan-ASA) assay and denaturing HPLC assay to detect the mutation of CYP2C9 gene. A single base mutation in the CYP2C9 gene was found in one of thirty subjects in the healthy population. We suggest that this mutation in the CYP2C9 gene may be related to diclofenac-induced influenza-virus-associated encephalopathy. PMID- 11925851 TI - [Drug-drug interaction in pharmacogenetics and pharmacogenomics]. AB - Drug metabolism and excretion is composed of four steps: Absorption, distribution, metabolism and excretion. The four steps are often abbreviated as ADME. Drug-drug interaction may occur at each step of ADME. Reported examples of drug-drug interaction occur mainly at the level of "drug metabolizing enzymes(DME)". The mechanisms of drug-drug interaction are: 1) Competitive inhibition of DME, 2) Destruction or irreversible inhibition of DME, 3) Induction of DME. Co-administration of 5-fluorouracil and sorivudine resulted in severe gastrointestinal and bone marrow toxicities. The toxicity is due to irreversible inhibition of dihydropyrimidine dehydrogenase by a sorivudine metabolite, which plays a role in detoxification of 5-fluorouracil. However, there is an example of beneficial drug-drug interaction, where proton pump inhibitor, omeprazole, antibiotics, amoxicillin and clarithromycin, are co-administered for eradication of Helicobacter pylori. Omeprazole is metabolized by CYP2C19 and CYP3A4. In poor metabolizers of omeprazole, a higher area under the drug concentration curve(AUC) and higher efficacy are achieved as compared to extensive metabolizers of omeprazole. In this regimen, co-administration of clarithromycin which is metabolized by CYP3A4 effectively raises the AUC of omeprazole. Thus, this drug combination results in a beneficial drug-drug interaction. PMID- 11925852 TI - [Drug of abuse]. AB - Recent development of various dietary supplements after enforcement of "Dietary Supplement Health and Education Act of 1994 (DSHEA)" in the USA enabled better availability of the products through the Internet in Japan as well. Because of differences in the definitions of the term "dietary supplement" and drug control laws between the USA and Japan, health risks due to uncontrolled use of a drug based foreign dietary supplement without a medical doctor's advice, and side effects due to co-administration of any problematic supplements with prescription drugs has become a problem in Japan. Classes of typical dietary supplements, the method of distribution, and known problems during use or overuse of these products with prescription drugs are discussed. Several recent positive cases are known to be due to the use of contaminated food supplements, which were sold not only to athletes but also to the general public as memory enhancing or anti-aging drugs. These phenomena indicate that trends in drug use in sports and in society becoming increasingly similar. PMID- 11925853 TI - [Human gene tests as laboratory medicine in the post-genome era]. AB - Massive and systematic information on human genes and genome is accumulating with progress in the Human Genome Project. Information on the genome has a characteristic digital signal that is appropriate for high-throughput handling by computers. Therefore, the field of gene tests has a high affinity to the field of laboratory medicine. In addition, it is a important survival strategy for departments of laboratory medicine to remain at the cutting edge. We must establish an infrastructure for utilizing genome information in clinical medicine and must anticipate numerous issues. The following issues need to be addressed. 1. Online database of human gene tests (http://www.kuhp.kyoto u.ac.jp/idennet/DB/index2.html) 2. Anonymous handling of patients' samples 3. Establishment of independent Department of Clinical Genetics 4. Education of clinical geneticists 5. Quality control of human gene tests (especially germline mutation tests) 6. Education of technicians handling human gene tests. PMID- 11925854 TI - [cDNA microarray technology as a laboratory examination method: a gene expression profiling test for analysis of drug resistance in tumor cells]. AB - cDNA microarray technology permits the simultaneous measurement of the expressions of thousands of genes. The technology is now an indispensable research tool in molecular biology, and the challenge is its development and usage as clinical diagnostic tools. cDNA microarray can be used to identify gene expression profiles in tumor cells which correlate strongly with the treatment responsiveness such as drug resistance and clinical outcome of the disease despite similar phenotypes. For the introduction of cDNA microarray into laboratory examination, many issues need to be resolved. Design of a diagnostic array must be developed with defined sequences based on interpretation of huge quantities of data from experimental arrays to predict treatment responsiveness. Assay quality must be improved in terms of detection sensitivity, reproducibility, and linear dynamic range for RNA quantitation. Generally available instruments, which are much less expensive and more practical, need to be developed. Along with the improvement of the assay as a laboratory examination method, cDNA microarray will facilitate the integration of diagnosis and therapeutics, and the introduction of individual medicines. PMID- 11925855 TI - [Proteomics in clinical research: new approach of mass spectrometry]. AB - A proteome has been defined as the protein complement expressed by the genome of an organism, tissue, or differentiated cell. Knowledge of complete genome sequences has led to considerable effort being increasingly devoted to the large scale study of proteomes, that is, 'proteomics'. Commonly, two proteomes are compared by a substructive analysis in which differences due to drug treatment, culture conditions, genetic variations, or diseases can be observed. Two dimensional gel electrophoresis and mass spectrometry are commonly used for this purpose. We applied this approach to the analysis of vitreous humor(VH) proteins. Fifty-two different proteins were identified on silver-stained 2D-gel patterns with VH proteins obtained from diabetic retinopathy and macular hole. Thirty-five proteins, which have not reported in plasma, were found in VH. Pigment epithelium derived factor, which was reported to be a potent inhibitor of angiogenesis in cornea and vitreous was at a higher concentration in VH with diabetes than in that with macular hole. It is impressive that the inhibitor increases in the vitreous with proliferative angiogenesis. Unique applications in proteomics promise a bright future for molecular biology and hopefully for clinical chemistry. PMID- 11925856 TI - [Undecatungstophosphate sensitized strains of methicillin-resistant Staphylococcus aureus to beta-lactams]. AB - Previously, a factor (Factor T) was found in aged mixtures of tungstate and phosphate, which greatly sensitizes strains of methicillin-resistant Staphylococcus aureus to beta-lactams. Factor T was purified and identified as undecatungstophosphate([PW11O39]7-). Undecatungstosilicate([SiW11O39]8-), a compound closely related to undecatungstophosphate, showed a similar enhancing effect. Chemically, these compounds are classified as "polyoxotungstates", and it is expected that these tungsten compounds will be useful as a "tool" in laboratory tests: i.e., in screening media for highly resistant MRSA strains. They may be also useful to investigate the resistant mechanism of the bacterial cells. PMID- 11925857 TI - [Effect of aging on heart rate and blood pressure variabilities in healthy men and women]. AB - OBJECT: To clarify the effect of aging and gender on heart rate and blood pressure variability in healthy men and women. SUBJECT AND METHODS: One hundred ninety four healthy subjects aged from 20 to 70(97 men and 97 women) were examined. Heart beat and blood pressure during 5 minutes in the supine and standing positions were measured under a respiration rate of 15/min. Blood pressure and heart were measured with Finapres and Biotop, respectively. Data entry and spectral analysis were carried out by the Fast Fourier Transform(FFT) method with BIMUTAS-II, after correcting 256 data with Hanning window function and resampling at 200 Hz. RESULT AND DISCUSSION: High frequency power(HF: 0.15 0.50 Hz) of heart rate variability decreased with age in both men and women, Low frequency power (LF: 0.05-0.15 Hz) of systolic blood pressure(SBP) variability in the standing posture decreased with age in men. The HF of SBP variability in the standing posture was decreased with age in both men and women. These findings suggest that aging and gender affected heart rate and blood pressure variability. PMID- 11925858 TI - [Comparative study of T-wave alternans, QT c dispersion and late potential for predicting ventricular tachycardia in patients with ischemic heart disease]. AB - Ventricular tachycardia(VT) is well known as the life-threatening arrhythmia. It would be important for predicting the risk of VT to prevent sudden death caused by VT after myocardial damage such as old myocardial infarction and dilated cardiomyopathy. In this study, we examined late potential(LP), TWA alternans(TWA), and QTc dispersion(QTcd) measured by Holter ECG, in 21 patients with old myocardial infarction(OMI) and 21 patients with dilated cardiomyopathy(DCM), and evaluated these parameters in relation with the occurrence of VT on these patients. The sensitivities of LP, TWA, and QTcd in patients with OMI in relation with VT were 82%, 73%, and 82%, respectively, and those in patients with DCM were 82%, 93%, and 73%, respectively. The specificities of LP, TWA, and QTcd in patients with OMI in relation with VT were 100%, 40%, and 100%, respectively, and those in patients with DCM were 100%, 30%, and 100%, respectively. The data demonstrated that LP and TWA were the useful predictive parameters for the VT on myocardial damage such as OMI and DCM. PMID- 11925859 TI - [Correlation between expression of MMP-7 in gastric submucosal invasive carcinoma and lymph node micrometastasis]. AB - To clarify the mechanism of tumor metastasis, lymph nodes micrometastasis and expression of MMP-7, which is related to lymph node metastases, were investigated in 45 submucosal, invasive gastric carcinomas. Metastases to lymph nodes were detected in 12 of 45 cases(26.7%) by HE stain. In 13 of 45 cases (28.9%), only micrometastases were detected by immunostaining with anti-cytokeratin antibody. The incidence of micrometastases was higher in poorly-differentiated carcinoma than well-differentiated carcinoma. Expression of MMP-7 was higher in metastasis positive cases than in negative cases. In poorly-differentiated cases, expression of MMP-7 was associated with micrometastasis. In conclusion, expression of MMP-7 may play an important role not only in tumor metastasis but in micrometastasis to lymph node especially in poorly-differentiated cases. PMID- 11925860 TI - [Improvement of total prostate specific antigen(PSA) assay from the "Skewed molar response" to the "Equimolar-response" by utilizing anti free-PSA monoclonal antibodies]. AB - Prostate specific antigen(PSA) is widely used as a marker for screening and monitoring prostate cancer. However, there are several assay methods, which often give different PSA values for the same patient sample. A possible cause of these discrepancies might be the varied immunoreactivity with free-PSA and complex-PSA among methods. This study revealed that pre-treatment of samples with anti-free PSA monoclonal antibodies improved polyclonal antibodies-based non equimolar PSA assay to equimolar assay. This might be achieved by that the monoclonal antibodies bind to free-PSA and then change its reactivities with polyclonal antibodies to those similar to complex-PSA. PMID- 11925861 TI - [Helicobacter pylori infection and gastric carcinogenesis]. PMID- 11925862 TI - [Colon cancer and COX-2]. PMID- 11925864 TI - [A case of gastric carcinoids associated with type A gastritis and gastric cancer]. PMID- 11925863 TI - [Relationship between location of stress erosive gastritis and brain damage in resuscitated patients]. AB - Patients after resuscitation from cardiopulmonary arrest often show stress erosive gastritis. This study investigated the relationship between the location of gastric mucosal injury and the degree of brain damage. Forty-five resuscitated patients with gastrointestinal bleeding complications were enrolled and were examined by esophagogastric fiberscope after 72 hours of hospitalization. Their brainstem and cerebral functions were evaluated brainstem auditory evoked potential (BAEP) and electrical encephalogram (EEG), respectively. Thirty patients showed complications with acute gastric lesions. Ten patients had gastric mucosal injury in the antrum and they all showed a good response for BAEP (I, III and V waves were positive). In contrast, patients without antral gastric mucosal lesions showed poor response for ABR (defect of III and V waves) and EGG (Hockerday Grade III or IV). These results indicate that fair brainstem function is necessary for stress erosive gastritis in gastric antrum. PMID- 11925865 TI - [A case of AFP and PIVKA-II producing gastric carcinoma forming a massive tumor embolus in the portal vein]. PMID- 11925866 TI - [A case of neuroendocrine carcinoma of the rectum]. PMID- 11925867 TI - [A case of non-steroidal anti-inflammatory drug-induced colitis with granulomas]. PMID- 11925868 TI - [A case of solitary adrenal metastasis from rectosigmoidal carcinoma]. PMID- 11925869 TI - [A case of liver abscess with particular pathological features due to Alcaligenes xylosoxidans infection]. PMID- 11925870 TI - [A case of segmental primary sclerosing cholangitis associated with early cholangiocarcinoma--a review of the Japanese literature]. PMID- 11925871 TI - [Hypercalcemia mediated by parathyroid hormone-related protein in the blastic phase of chronic myelogenous leukemia]. AB - A 45-year-old man with chronic myelogenous leukemia (CML) in the accelerated phase was admitted to our hospital because of lower back pain and hypercalcemia. On admission, he was confused and found to have massive splenomegaly. The hypercalcemia and splenomegaly improved significantly after administration of incadronate, hydroxyurea, vincristine and prednisolone. Splenomegaly recurred after cessation of the chemotherapy, and examination of the peripheral blood showed 31% blasts, positive for both CD13 and CD33, on which basis myeloid blastic transformation was diagnosed. Vindesine, cytarabine and prednisolone were administered, and the splenomegaly improved again. On admission, when the patient's serum calcium level was 16.0 mg/dl, his serum parathyroid hormone related protein (PTHrP) level was elevated to 118.3 pmol/l. Furthermore, RT-PCR analysis revealed that the patient's CML cells expressed PTHrP mRNA, and a high level of PTHrP was detected in the supernatant of cultured mononuclear cells derived from the patient's peripheral blood. These findings indicated that the hypercalcemia was due to production of PTHrP by the leukemic cells. Several cases of PTHrP. mediated hypercalcemia associated with CML have been reported previously, and are reviewed here. PMID- 11925873 TI - [Successful treatment of myeloid/natural killer cell precursor acute leukemia with allogeneic peripheral blood stem cell transplantation]. AB - We report a case of myeloid/NK cell precursor acute leukemia, which was successfully treated with allogeneic peripheral blood stem cell transplantation (allo PBSCT). A 31-year-old woman was admitted to our hospital with general fatigue, anorexia and leukocytosis. Bone marrow aspiration showed infiltration of many atypical blasts. She was diagnosed as having myeloid/NK cell precursor acute leukemia by morphological and immunohistochemical analysis. Complete remission was achieved by induction chemotherapy, but as myeloid/NK cell precursor acute leukemia is reported to have an extremely poor prognosis due to frequent relapse, the patient underwent allo PBSCT from her HLA-identical father, together with a myeloablative conditioning regimen. She suffered several transplantation-related complications including acute graft versus host disease (grade II) and ischemic enterocolitis associated with thrombotic microangiopathy, but these were overcome by supportive therapy. She was discharged on day 168 after allo PBSCT, and so far there has been no evidence of relapse during a follow-up period of 15 months. PMID- 11925872 TI - [Nonsecretory primary plasma cell leukemia successfully treated with VAD and MP therapy]. AB - A 78-year-old woman was admitted to our hospital because of lumbago and appetite loss. Blood analysis revealed anemia, hypercalcemia and circulating plasma cells. Bone marrow aspiration showed an elevated ratio (43%) of plasma cells, which expressed CD38 in the absence of CD19 and CD56 expression. Spinal MR imaging revealed multiple compression fractures and suggested diffuse invasion of plasma cells into the spinal bodies. No M-protein was detectable in serum or urine by immunoelectrophoresis and immunofixation, but cytoplasmic M-protein (IgG-kappa) was detected by enzyme antibody staining. On the basis of the history and data, nonsecretory primary plasma cell leukemia was diagnosed. First, the patient was given modified VAD therapy (vincristine, doxorubicin, and prednisolone) and complete remission was obtained. Then MP therapy (melphalan and prednisolone) was instituted, and remission has since been maintained for 11 months. Like many other cases of primary plasma cell leukemia, this case suggests that CD56 may act as an adhesion molecule between neoplastic plasma cells and bone marrow stromal cells. Our experience with this exceedingly rare case suggests the superiority of combination chemotherapy as an induction therapy and the effectiveness of MP therapy as maintenance therapy for this disease. PMID- 11925874 TI - [Vitamin K2 therapy for myelodysplastic syndrome]. AB - Vitamin K2 is reported to induce apoptosis or differentiation of leukemic cell lines in vitro. We administered a vitamin K2 analog, menatetrenone, at 45 mg daily to 23 patients with myelodysplastic syndrome (MDS): 13 patients with RA, 2 with RARS, 6 with RAEB and 2 with RAEB-T. Good response (GR) and partial response (PR) were defined as an increase of hemoglobin concentration exceeding 2 g/dl and 1-2 g/dl without transfusion, respectively. Six of the RA patients showed improvement of anemia (GR, 3 patients; PR, 3 patients). RA patients who did not have a hypocellular bone marrow and were transfusion-independent tended to be responsive to vitamin K2 therapy in combination with vitamin D3 or anabolic steroids. No adverse effect of vitamin K2 was observed, and the time required to obtain the hematological response was short, being 3 months on average. We believe that vitamin K2 therapy has potential as a treatment for patients with MDS. PMID- 11925875 TI - [Systemic capillary leak syndrome presenting remarkable erythrocytosis]. AB - Systemic capillary leak syndrome (SCLS) is a disorder characterized by hypotension, edema, and an increased hematocrit (Ht) due to sudden leakage of plasma into the extravascular space through some unknown mechanism, in which monoclonal gammopathy is observed. A 30-year-old man visited our emergency department because of abdominal pain, and was admitted to our hematology department because of a markedly increased hemoglobin concentration reaching 26.2 g/dl. The polycythemia was thought to be pseudo-polycythemia due to hemoconcentration, and we diagnosed the patient as having SCLS based on the triad of increased hematocrit, whole-body edema which was especially marked in the lower extremities, and monoclonal gammopathy. The patient recovered after administration of extracellular fluids and albumin, but the attacks recurred. Prophylaxis with terbutaline sulfate, theophylline and corticosteroid reduced the frequency of severe attacks. Because there is possibility that patients with SCLS may be admitted to hematology departments due to severe erythrocytosis, we report this case to increase the awareness of hematologists that SCLS is one of the important differential diagnoses of erythrocytosis. PMID- 11925876 TI - [CD5+ CD10+ extranodal B-cell lymphoma with tetraploid chromosomal abnormality]. AB - An 84-year-old man was admitted with pleural lymphomatous effusion and bone marrow infiltration. The abnormal cells from the effusion showed abundant basophilic cytoplasm, large atypical nuclei, and small nucleoli with frequent mitosis. The abnormal cells were found to be CD5+, CD10+, CD19+, and CD20+ by flow cytometry, and had clonal rearrangements of the IgH-JH gene, indicating the existence of a clonal B-cell population. No bcl-1, bcl-2, bcl-6, or c-myc rearrangement was found. Neither human herpesvirus 8 (HHV-8) nor Epstein-Barr virus was detected in the abnormal cells. Tetraploid chromosomal abnormality was present. After administration of prednisolone, transient disappearance of the effusion was obtained. PMID- 11925877 TI - [The 43rd annual meeting of the Japanese society of clinical hematology education lectures: biological activities of IL-18 and its involvement in disease pathogenesis]. PMID- 11925879 TI - [Transfusion and its adverse effects in patients with hematological disorders]. PMID- 11925878 TI - [Clinical disorders of blood coagulation and fibrinolysis]. PMID- 11925880 TI - [Current status of peripheral blood stem cell transplantation: comparison with bone marrow a shematopoietic stem cell graft]. PMID- 11925881 TI - [Current clinical problems in non-myeloablative reduced-intensity hematopoietic stem cell transplantation]. PMID- 11925882 TI - [Immunological tolerance and rejection after hematopoietic stem cell transplantation]. PMID- 11925883 TI - [Development of immunotherapy using allogeneic antigens for hematological malignancy]. PMID- 11925884 TI - [Adult-onset Still's disease accompanied by hemophagocytic syndrome at onset]. AB - A 43-year-old woman was admitted for examination of fever, an elevated transaminase level, LDH, skin eruption, sore throat and bicytopenia. As bone marrow examination revealed an increased proportion of histiocytes and active phagocytosis, hemophagocytic syndrome (HPS) was diagnosed. After admission, the peripheral blood counts recovered spontaneously and the HPS subsided, but other symptoms persisted and the neutrophil count increased. At this time, we diagnosed the patient as having adult Still's disease. All the symptoms disappeared after administration of prednisolone. The markedly increased concentrations of TNF alpha and IFN-gamma in the peripheral blood at the time of HPS declined gradually, and the IL-6 concentration increased at the time of diagnosis of Still's disease. However, all of these concentrations normalized after administration of prednisolone. As HPS and Still's disease have a common etiology, and each shows high concentrations of IFN-gamma, IL 6 and TNF-alpha, the symptoms are similar in both diseases. In particular, a relationship between HPS and high concentrations of TNF-alpha and IFN-gamma is suspected. PMID- 11925885 TI - [The effect of normobaric oxygen inhalation and halothane anesthesia on the level of interstitial striatal dopamine of rats using in vivo microdialysis study]. AB - We investigated the effect of normobaric oxygen inhalation on the level of interstitial dopamine and its metabolites in in vivo brain striatum of awake, free moving rats using microdialysis techniques. Rats were implanted a microdialysis probe to the right striatum of the brain and administered pure oxygen for 1 hour, and dialysates from the probe were examined every 20 minutes by HPLC. Normobaric oxygen inhalation reduced the amount of dopamine derived from the dialysate and increased that of metabolites. Halothane anesthesia with oxygen showed a slight effect on the changes induced by oxygen inhalation, whereas halothane significantly increased the level of dopamine metabolites. Pretreatment with glutathione failed to prevent increase of dopamine metabolites. We hypothesized that oxygen inhalation and halothane anesthesia might increase the level of dopamine metabolites in different mechanisms, and the changes induced by oxygen inhalation might not be the result of simple oxidative stress in rat striatum. PMID- 11925886 TI - [Arterial Mg2+ concentrations and ECG. Changes in patients with cerebral aneurysms]. AB - Arterial Mg2+ concentrations were analyzed in 108 patients with cerebral aneurysms. There was no difference in Mg2+ concentrations between patients with ruptured and unruptured aneurysms. Mg2+ concentrations in patients with ST depression or elevation were lower than in those without ST changes (0.34 +/- 0.05 vs. 0.38 +/- 0.07 mmol.l-1). Decreased arterial Mg2+ could be caused by cardiac ischemia rather than subarachnoid hemorrhage. For prevention of fatal arrhythmias, perioperative brain protection and ameliorating vasospasm, correcting the depressed Mg2+ concentrations is suggested. PMID- 11925887 TI - [Management of intraoperative anticoagulantless CHD with EVAL-dialyzer in four patients on maintenance hemodialysis]. AB - It is difficult to correct fluid balance during operation in patients under hemodialysis. We managed intraoperative anticoagulantless continuous hemodialysis (CHD) with ethylene vinyl alcohol copolymer (EVAL) dialyzer in four patients on maintenance hemodialysis. All patients could be managed satisfactorily with no coagulation. Only in the first patient, we used nafamostat mesilate to avoid coagulation in the dialyzer. In this patient, activated clotting time (ACT) was 151 seconds. Their recovery from anesthesia was good, and they showed no complications perioperatively. This is a good method to manage these patients during operation, because EVAL dialyzer is cheap and we need not take care of hemorrhage. Using a small amount of nafamostat mesilate may be useful to avoid coagulation. PMID- 11925888 TI - [Severe hypotension after removal of pheochromocytoma in a pediatric patient]. AB - An 8-year-old male patient with pheochromocytoma pretreated with prazocin, carteolol, and enarapril received sevoflurane general anesthesia with fentanyl. After removal of the tumor, blood pressure and urine volume decreased and ST segment in ECG was depressed. We administered dopamine and norepinephrine to increase blood pressure, but pulmonary edema became aggravated postoperatively. He was treated with noninvasive pressure support ventilation with face mask for 4 hours which was effective. His postoperative recovery was uneventful thereafter. We hypothesized that his hemodynamic deterioration was caused by sudden decrease in serum level of norepinephrine after the tumor removal, and the responsiveness to catecholamines was depressed by long exposure to norepinephrine. We conclude that we should use norepinephrine for anesthetic care even in pediatric patients as in adults. PMID- 11925889 TI - [Anesthesia for partial pulmonary resection in a postpneumonectomy patient]. AB - We experienced anesthesia for partial pulmonary resection in a postpneumonectomy patient. A 59 year-old woman underwent left upper lobectomy 5 years earlier and left lower lobectomy 2 years earlier for lung cancer, respectively. Right segmental pulmonary resection was proposed for lung cancer. Preoperative examinations revealed decreased FVC (1.81 l) and FVC1 (1.57 l), and increased PaCO2 (48.4 mmHg). The patient was monitored with EKG, direct arterial pressure, central venous pressure, pulse oxymetry and expired gas analysis. Anesthesia was induced with thiopental and fentanyl, and maintained with sevoflurane, propofol and supplemental fentanyl after the establishment of epidural anesthesia. Dopamine was continuously infused due to decreased blood pressure. Then hemodynamics was stable. Operative procedure was protective. Therefore SPO2 was kept above 98% during the operation. Partial resection was performed instead of segmental resection to avoid deteriorating the postoperative quality of life. The postoperative course was uneventful. PMID- 11925890 TI - [Acute renal failure in an infant attributable to arterial cannula malposition during cardiopulmonary bypass via ministernotomy]. AB - A 6-month-old female (4.9 kg) with multiple congenital heart lesions underwent intracardiac repair with the aid of cardiopulmonary bypass (CPB) through a lower half sternotomy. Aortic cannulation, venous cannulation, and cardioplegia cannula insertion were all accomplished through the ministernotomy. During the CPB, in spite of a high perfusion flow rate (182 ml.kg-1.min-1), the systemic arterial pressure was persistently low (mean values = 25-35 mmHg) and the urine output was greatly reduced (< 1 ml.hr-1). In addition, inappropriate increases in the arterial inflow line pressure were recognized. Since abutment of the cannula tip against the aortic intima was suspected, several attempts were made to correct its malpositioning. During the CPB, hemolysis was also found in the mixed venous blood. Since the oliguria and resultant hyperkalemia persisted after weaning from the CPB, peritoneal dialysis was introduced immediately after the surgery. Her renal function gradually recovered postoperatively, and she was finally weaned from the peritoneal dialysis on the 13th postoperative day. Although the ministernotomy has been proposed to be a safe approach for most of cardiac surgeries, it appears to increase the risk for arterial cannula malposition as compared to the standard full-length sternotomy in small pediatric patients. PMID- 11925891 TI - [Recurrent cardiac arrest after splenectomy in a patient with ITP and diabetes mellitus]. AB - We report a case of 58-year-old man who had repeated cardiac arrests on the first post-operative day. The patient underwent splenectomy due to ITP (idiopathic thrombocytopenic purpura). He also had diabetes mellitus and nephrotic syndrome. There was no abnormal finding at the preoperative examination, except bleeding time of 6 minutes. The operation was finished without complications under general anesthesia. Midnight on the day of surgery, the first cardiac arrest occurred, and lasted for about 10 seconds. He recovered soon from the incident, but at 6 o'clock next morning, he developed severe bradycardia and cardiac arrest. He recovered again, but around 10 o'clock, he developed bradycardia and arrest again, and fell into a fit of convulsions and lost his consciousness. Again he recovered soon and no bradycardia and cardiac arrest occurred after this episode. Two years later, he was scheduled for vitrectomy due to diabetic retinosis. There were a few PACs and PVCs in his Holter-ECG, but no typical bradycardia and ST changes. During the operation, we injected atropine sulfate, dopamine hydrochloride and bucladesine sodium to increase his heart rate above 60 per minute. The operation was finished smoothly and there was no trouble perioperatively. A year later, he also underwent bilateral cataract extraction under local anesthesia without any troubles. PMID- 11925892 TI - [Three times of anesthetic management in a patient with myotonic dystrophy]. AB - A 38-year-old female, at 38-week gestation, was scheduled for cesarean section under epidural anesthesia. After the delivery, it was found that she had been diagnosed as myotonic dystrophy by the other physician and the neonate was a floppy infant indicating hereditary neuromuscular diseases. In her case, myotonic dystrophy had not been advanced and symptoms had been mild. We previously had given her general anesthesia for two times with nitrous oxide, isoflurane and vecuronium for her to undergo emergency operations, left salpingectomy under laparoscopy due to unruptured tubal pregnancy at 34 years of age and cesarean section due to liver function disorder indicating HELLP syndrome at 36 years of age. Although many problems have been described about the perioperative management in patients with myotonic dystrophy, she was safely managed for each operation. PMID- 11925893 TI - [Intraoperative massive pulmonary hemorrhage due to pulmonary aspergillosis]. AB - A 16-year-old girl had suffered from chronic graft versus host disease (GVHD) caused by peripheral blood stem cell transplantation (PBSCT) after chemotherapy for neuroblastoma and pulmonary aspergillosis of the right upper lobe. She presented with hematemesis and underwent upper gastrointestinal endoscopy under general anesthesia. At the end of the examination, massive pulmonary hemorrhage occurred suddenly. A double lumen endobronchial tube was inserted for unilateral ventilation in order to control hemorrhage, and right pulmonary hemorrhage was found. Pulmonary scintigram and angiography could not demonstrate the bleeding site, and we suspected that pulmonary hemorrhage had been caused by pulmonary aspergillosis because aspergillus is known to have pathologically invasive character to the adjacent tissue and blood vessels. Despite right pneumonectomy was performed to control pulmonary hemorrhage, she died five days later from multiple organ failure. This case suggests that immediate unilateral ventilation is useful for the isolation of the bleeding lung when pulmonary hemorrhage is massive and we should know the risk of pulmonary hemorrhage in patients with pulmonary aspergillosis. PMID- 11925894 TI - [A case of postoperative acute myocardial infarction due to the interruption of anticoagulant therapy]. AB - A 67-year-old male patient was scheduled for stripping of a lower limb varix. He had received long-term anticoagulant therapy using oral warfarin because of implantation of an aortic mechanical heart valve. Warfarin therapy was discontinued 3 days before surgery to prevent excessive bleeding during the perioperative period. Anesthesia was maintained with sevoflurane and nitrous oxide in oxygen and combined with continuous epidural block. The operation was uneventful. Six hours after surgery, an electrocardiogram demonstrated ventricular fibrillation, followed by respiratory arrest. Cardiopulmonary resuscitation was started immediately, and normal sinus rhythm was restored. Coronary angiography revealed significant stenosis of coronary arteries. Percutaneous transluminal coronary angioplasty was therefore performed. Acute myocardial infarction caused by the interruption of anticoagulant therapy was suspected. PMID- 11925895 TI - [Repeated coronary artery spasm under general anesthesia]. AB - A 52-year-old man without history of angina pectoris underwent emergent abdominal surgery for acute abdomen. General anesthesia was maintained with isoflurane, nitrous oxide and fentanyl. The depth of anesthesia during surgery was considered adequate for the stimuli, but the patient developed ECG abnormality repeatedly finally followed by short run. After anesthesia, we re-examined the recorded ECG and coronary spasm was strongly suspected. Fatal arrhythmia might be avoidable if the prior abnormality in ECG is properly assessed. PMID- 11925897 TI - [Anesthesia in a patient with relapsing polychondritis]. AB - A 46-year-old female with relapsing polychondritis (RP) was scheduled for surgical repair of saddle-nose and peri-chondritic ear. RP is a rare systemic, inflammatory, and destructive disease of the cartilaginous structures leading to multiple functional disorders in the affected organs. Preoperatively, her chest radiogram of the trachea showed narrowing in diameters which altered about 2 mm with expiration and inspiration. Propofol administered intravenously was given as induction and anesthesia was maintained with nitrous oxide-oxygen-sevoflurane with spontaneous ventilation using laryngeal mask airway. The operation was over uneventfully. Anesthesiologic management requires careful preoperative evaluation of vital organ functions, in particular, respiration. PMID- 11925896 TI - [The effect of dopamine on serum lipid concentration after propofol administration]. AB - Propofol in emulsion formulation is widely used for operation and sedation in ICU. We retrospectively investigated the effect of dopamine on post-operative serum lipid concentrations after propofol administration. Twenty three patients with ASA physical status I or II scheduled for elective operations were enrolled in this study; 15 patients in the non-dopamine administration group (Group P) and 8 patients in the dopamine administration group (Group Dopa-P). We measured triglyceride (TG), total cholesterol (T-chol) pre-operatively, post-operatively and on post-operative day 1 and AST, ALT preoperatively and on post-operative day 1. Serum TG concentration increased significantly in post-operative measurements in Group-P (P < 0.05). But there was no significant change in TG in Group Dopa-P. Serum T-chol concentration decreased significantly post-operatively and on post operative day 1 in both groups (P < 0.05). Serum AST and ALT concentrations increased significantly on post-operative day 1 (P < 0.05). With 10% fat infusion rates below 0.1 mg.kg-1.hr-1 (equal to propofol 10 mg.kg-1.hr-1), serum lipid concentration did not increase. But our results suggest that the serum TG concentration may increase significantly post-operatively after intra-operative propofol administration and dopamine may decrease serum TG level. PMID- 11925898 TI - [The development of methicillin-resistant Staphylococcus aureus sepsis in a patient with herpes zoster during treatment with continuous epidural infusion]. AB - A 79-year-old man with herpes zoster was referred to our hospital for pain control. He was a survivor of the atomic bombing of Hiroshima, and had a history of cerebral infarction and hypertension. A cervical epidural catheter was placed for continuous analgesic infusion. After 20 days of catheterization, he gradually developed a high fever and confusion, and complained of nausea and headaches. An urgent blood examination revealed a white blood cell count of 15,200 mm-3 and a C reactive protein of 32.4 mg.dl-1. The catheter was removed and antibiotic therapy was started. Repeated magnetic resonance imaging could not confirm epidural abscess formation. The bacterial culture of the cerebrospinal fluid was negative, but the cultures of the blood, the catheter tip, and the nasal cavity swab were positive for methicillin-resistant Staphylococcus aureus. Although intravenous vancomycin was administered, systemic inflammation persisted. The patient consecutively suffered varied disorders such as acute renal failure, disseminated intravascular coagulation, and gastrointestinal bleeding. Although symptomatic treatment had been prolonging his life, 58 days after the catheter removal, the patient suddenly developed cerebellopontine infarction, which made mechanical ventilation necessary. He remained unconscious until his death 117 days after the catheter removal. We discussed the possible pathogenetic mechanisms of the present case. PMID- 11925899 TI - [Survey of smoking status among preoperative patients and characteristics of smokers]. AB - Smoking is an independent risk factor for both pulmonary and nonpulmonary perioperative complications. For safer anesthetic management, it is important to encourage and support the cessation of smoking in the preoperative period. As a first step to design a preoperative smoking cessation program, we conducted a survey of preoperative patients about smoking status and characteristics of smokers at an outpatient clinic for an anesthesiology department. The percentages of male and female smokers were 42% and 19%, respectively. Percentages of preoperative smokers considered to be in the preparation stage of smoking stage were 26% in male and 19% in female, as compared to 3% in male and 5% in female in general smokers. Twenty four % of both male and female patients were strongly committed to achieving smoking cessation. These trends in smoking characteristics indicate that preoperative smokers are more likely to quit smoking without heavy support and encouragement than general smokers. In designing a preoperative smoking cessation program, these results must be taken into consideration. PMID- 11925900 TI - [Distribution of an explanatory note concerning the risks of anesthesia prior to preanesthetic evaluation]. AB - To make the public aware of the risks of anesthesia, we prepared an explanatory note composed of 634 Japanese characters or of 248 English words. The incidences of fatal anesthetic complications over a five-year period in 2,358,642 anesthetics in 741 Certified Training Hospital belonging to the Japanese Society of Anesthesiologists were cited in the explanatory note. Patients were asked to read and sign the explanatory note prior to agreeing to surgery. After patients having received this general information about surgical and anesthesia risks, surgeons then apply to the Department of Anesthesiology for their anesthetic management. Thereafter, responsible anesthesiologists visit and evaluate patients, and explain common as well as specific anesthesia risks to each patient. A survey by mailing questionnaires regarding this explanatory note and anesthesia risks sent to patients, who had read and signed the explanatory note, revealed that the patients were generally satisfied with the content of the explanatory note. This system may help patients, surgeons and anesthesiologists to recognize anesthesia risks on the same basis. PMID- 11925901 TI - [Introduction of the electronic anesthesia record keeping system]. AB - Based on five-year experience, we have evaluated the electronic anesthesia record keeping system (DS 5300 OR, Fukuda Denshi, Tokyo), which is the first commercially available system combined with an anesthesia monitor in Japan. Although keying in of text with a keyboard on the screen remains to be improved, the user interface of touch screens is easy to use. It seems to be a great advantage that physiologic variables are automatically and accurately captured very minute. The record is more complete, legible, and accurate than a hand written record. Both long-term storage and search for the data are much easier. The investment in this system is affordable even for small hospitals. Although there might be some areas in which improvement is needed, we are satisfied with its usefulness and reliability. PMID- 11925902 TI - [Anesthesiology in People's Republic of China in the year 2001]. AB - In June 2001, we joined the regional anesthesia & Pain Medicine Delegation to the People's Republic of China. We visited 10 large hospitals in Beijing, Shanghai and Guiyang. We observed clinical anesthesia in operating theater and TCM (Traditional Chinese Medicine) in out-patient clinics. Acupuncture anesthesia was not popular and less frequently used for clinical anesthesia ranging 0 to 10%. Many surgical cases were done by Western medicine similar to Japan and USA. General anesthesia machines were imported from Germany and USA. All endo-tracheal tubes and catheters were made in China. But they lack the organization of anesthesia societies and system of specialization in the field of anesthesia. PMID- 11925903 TI - [Molecular mechanism of glucocorticoid action]. PMID- 11925904 TI - [A multicenter cross-sectional study on the health related quality of life of patients with rheumatoid arthritis using a revised Japanese version of the arthritis impact measurement scales version 2 (AIMS 2), focusing on the medical care costs and their associative factors]. AB - OBJECTIVE: To survey the actual conditions of medical care costs incurred by Japanese patients with rheumatoid arthritis, and to investigate impacts of health status, demographic and socioeconomic factors, clinical and laboratory measures, and medications on disease costs. METHODS: Self-reported health status questionnaires of the revised Japanese version of AIMS 2, and reports on out-of pocket medical care costs were collected from 1471 patients with classical and definite rheumatoid arthritis recruited through the arthritis study group of Ministry of Public Health and Welfare consisting of eleven arthritis centers across the country during three months from September, 1994. Impacts of health status and other demographic and clinical factors on medical care costs were statistically analyzed by using chi-square tests for categorical variables and Spearman's rank correlation analysis for numerical variables. RESULTS: 1. Averaged out-of-pocket medical care costs for RA patients was estimated at yen 25,225 ($253.5) per person-month in 1994, in which direct medical care costs accounted for 53.9% and indirect medical care costs accounted for 46.1% of the total. Averaged substantial direct medical care costs including the costs covered by the public health insurance in addition to out-of-pocket costs was estimated at yen 512,000 ($5,140) per person-year based on the averaged 11.8% self-pay rate of the public health insurance in 1994. 2. The distribution curve of the total out-of-pocket medical care costs was highly skewed. Averaged total medical care costs in the 90th, 95th, and 100th percentiles were 4.5, 8.1, and 48 times as large as those in the median percentile, respectively. 3. Out-of-pocket direct medical care costs totaled in the top 1 and 5 percentiles reached 26.6%, and 57.6% of those in whole patients, respectively. 4. Variables most strongly related to the total out-of-pocket medical care costs were work disability in AIMS health status scales, followed by physical disability, rate of functional decline, pain, affect, daily dose of oral prednisolone, global assessment by physician, joint counts, blood levels of CRP, ESR, grip strength, blood concentrations of hemoglobin, age, Steinbrocker's class, sex, and medications, in this order. 5. There was a trend of increase in number of cases of male, middle aged, with lower levels of formal education and annual income, longer disease duration of 20 years or more, and single (male) or separated (female) in marital status, as the total out-of-pocket medical care costs increased. 6. The average rate of missed days due to illness to whole working days of RA patients was estimated at 21.9%. With increase in the rate of missed days, the annual income of RA patients decreased, indicating that the lower annual income of RA patents possibly resulted from their work disabilities. Based on the rate of missed days, the average earning loss due to the illness in RA patients was estimated approximately at yen 650,000 ($6,540) perperson-year, which was equivalent to 1.3 times the average direct medical care costs for RA per person-year. CONCLUSION: The costs of RA were strongly related to work disability and other health status as represented by AIMS-HRQOL scores. A small number of patents severely disabled shared a disproportionately large part of medical care costs. To reduce the costs, the measures to prevent the development of disability are most important. PMID- 11925905 TI - [Possible roles of TNF family cytokines in bone destruction of rheumatoid arthritis]. PMID- 11925906 TI - [How Japanese clinicians treat rheumatoid arthritis (RA): a survey at the 44th annual meeting of Japan Rheumatism Association in 2000]. AB - OBJECTIVE AND METHODS: To gain insight into the current management of RA in Japan, a survey was done at the 44th annual meeting of Japan Rheumatism Association in 2000. RESULTS: Four hundred and ninety clinicians responded to our survey, including 212 internists and 252 orthopedists, respectively. In the management of RA, there was a wide difference between their present behaviors and the ideal way they believe. In particular, an appropriate communication between internists and orthopedists was lacking. There was a wide variation in their evaluations and prescribing behaviors. The treatment of RA in Japan turned out to be unique compared with those in the US and Europe. Prescriptions with little evidence were not unusual. CONCLUSIONS: There was a great variation among Japanese clinicians in the management of RA. These results might be associated with poor outcomes of RA patients in Japan. Dissemination of information concerning the management of RA, such as guidelines, as well as the establishment of a better health care system is necessary. PMID- 11925907 TI - [Difference in the management of rheumatoid arthritis (RA) by internists and orthopedists: a survey at the 44th annual meeting of Japan Rheumatism Association in 2000]. AB - OBJECTIVE AND METHODS: One of the reasons of the wide variation in the management of RA in Japan might be due to the different behaviors between internists and orthopedists. To investigate this possibility, a survey was done at the 44th annual meeting of Japan Rheumatism Association in 2000. RESULTS: Four hundred and ninety clinicians responded to our survey, including 212 internists and 252 orthopedists, respectively. In the management of RA, there was a wide difference between two groups. In particular, although both agreed that an appropriate communication between internists and orthopedists is necessary, half of the orthopedists answered that they don't refer their patients to internists. There was a wide variation in their ways of evaluations and prescribing behaviors. In particular, the use of DMARDs, both as monotherapy and combination, was significantly different between two groups. CONCLUSIONS: There was a great difference in the management of RA by internists and orthopedists. These results might be associated with the outcome of RA patients in Japan. Dissemination of information concerning the management of RA, such as guidelines, as well as the establishment of a better health care system is necessary. PMID- 11925908 TI - [Dynamics of matrix metalloproteinase (MMP)-13 in the patients with rheumatoid arthritis]. AB - OBJECTIVE: Matrix metalloproteinase (MMP) is a novel proteolytic enzyme that plays an important role in joint destruction in rheumatoid arthritis (RA). To elucidate the dynamics of MMPs in serum and synovial fluid, we measured the concentration and activity of MMP-1, -9, -13 in serum and synovial fluid of RA patients. Among them especially we focused on newly defined MMP-13 and compared with MMP-1 and MMP-9. METHODS: Serum, synovial fluid and synovial, and pannus tissues used in this study were obtained from RA patients. To compare the dynamics of each enzymic protein, we performed the following procedures: Firstly, we measured concentration of MMP-1, -9, -13 by using ELISA kit. Secondly, the activity of MMP-1, -9, -13 were also measured by using the MMP activity assay system. Then we obtained the activity ratio of each MMP from calculation of activity/concentration. We also examined the expression of MMP-13 in synovial tissues by immunohistochemical and in situ hybridization studies. RESULT: Concentration and activity levels of MMP-1, -9, -13 were significantly higher in RA serum and synovial fluid than in OA. Activity ratio of MMP-1, MMP-9 MMP-13 were 3.60 +/- 1.56, 1.03 +/- 1.75, 35.30 +/- 24.28 (ODA450/ng) in RA serum and 1.60 +/- 2.02, 3.97 +/- 14.83, 14.25 +/- 15.04 (ODA450/ng) in synovial fluid. In synovial and pannus tissues. MMP-13 positive cells were diffusely demonstrated by immunohistochemical and in situ hybridization studies. They were synovial lining cells, endothelial cells, fibroblasts, monocytes, osteoblasts, and chondrocytes. CONCLUSION: MMP-13 positive cells were diffusely presented in joint regions including synovial and pannus tissues. Although the concentration of MMP-13 was not so high, its activity ratio was elevated in serum and synovial fluid in the patients with rheumatoid arthritis. PMID- 11925910 TI - [A survey of the doses of methotrexate and salazosulfapyridine in Japanese patients with rheumatoid arthritis]. PMID- 11925909 TI - [A case of rheumatoid arthritis complicated with a pneumonitis during concomitant treatment with methotrexate and bucillamine]. AB - A 70-year-old female was diagnosed as having rheumatoid arthritis (RA) in 1971, which was then treated with steroid and nonsteroidal anti-inflammatory drugs. In 1999, after total replacement of her knee joint, 4 mg of methotrexate (MTX) per week was administered. Two months after the MTX administration, 200 mg of bucillamine per day was administered. On May 10, 2001, the patient was rushed to the hospital due to fever and difficulty in breathing. Chest X-ray and computed tomography (CT) revealed shadows of ground-glass-like opacity occurring sporadically in many places in the upper lung field bilaterally and interstitial shadows mainly on the lateral side of the lower lung field bilaterally. Instead of MTX and bucillamine, which were withheld, an MTX antagonist was administered and oxygen-supported therapy was performed; consequently, the patient recovered without the need to increase the amount of steroid. The percentage of lymphocytes in the broncholaveolar lavage fluid increased to 72%, and the CD4/CD8 ratio to 3.13. The level of serum KL-6 increased while that of serum SP-D returned to the normal level at different time. Following MTX and bucillamine administration, shadows of ground-glass-like opacity occurred sporadically in many places in the upper lung field bilaterally, which is not usually observed. It is suggested that such an unusual pulmonary disorder occurred due to concomitant use of drugs or other factors. PMID- 11925911 TI - [CT-guided transbronchial diagnosis using ultrathin bronchoscope for small peripheral pulmonary lesions]. AB - We performed computed tomography (CT)-guided transbronchial diagnosis on 23 patients, using an ultrathin bronchoscope (external diameter, 2.8 mm) for small peripheral pulmonary lesions (< or = 2 cm). The mean size of the lesions was 1.4 x 1.1 cm. After examination using a conventional bronchoscope (external diameter, 6.3 mm), an ultrathin bronchoscope and a biopsy apparatus were advanced to the lesion under CT and X-ray fluoroscopic guidance. The location of the biopsy apparatus at each lesion was confirmed by thin-section CT, and a biopsy was performed. The ultrathin bronchoscope reached the 5th-11th bronchus (mean, 7.1 +/ 1.5th), at a point 3.6 +/- 0.9 bronchi peripheral to the site reached using a conventional bronchoscope. In 20 of the 23 patients, the biopsy apparatus could be guided to the lesion. The diagnosis rate, by disease, was 81.8% (9/11) for lung cancer, 66.7% (2/3) for metastatic lung cancer, and 77.8% (7/9) for inflammation; the overall rate being 78.3% (18/23). CT-guided transbronchial diagnosis using an ultrathin bronchoscope is useful for diagnosing small peripheral pulmonary lesions because the bronchoscope can be readily inserted into peripheral areas and guided to lesions, and the site of sample collection can be accurately determined. PMID- 11925912 TI - [Cost-effectiveness analysis of ambulatory treatment for adult patients with community-acquired pneumonia: according to Japanese Respiratory Society guidelines]. AB - The Japanese Respiratory Society has recently formulated practice guidelines for the management of adult patients with community-acquired pneumonia. The guidelines recommend the use of various oral antibiotics at individual physicians' discretion. We compared the cost-effectiveness of amoxicillin/clavulanate (AMPC/CVA), azithromycin (AZM), clarithromycin (CAM), cefdinir (CFDN), levofloxacin (LVFX), and minocycline (MINO), when used on an ambulatory basis. We performed a formal cost-effectiveness analysis from the perspective of direct cost payers in the framework of the Japanese medical system. Outcomes considered were quality-adjusted life days (QALD), costs per patient, and incremental costs per quality-adjusted life year (QALY) gained. Under baseline conditions, the effectiveness of MINO, AZM, CAM, and LVFX were on a par and higher than that of AMPC/CVA or CFDN by 125-290.5 QALD. The least expensive antibiotic was MINO (55,070 to 59,208 yen), followed by AZM (56,049 to 60,188 yen), CAM (56,171 to 60,309 yen), LVFX (61,988 to 66,127 yen). AMPC/CVA (122,432 to 133,797 yen), and CFDN (123,375 to 134,649 yen). Thus, MINO, AZM, and CAM were cost-effective antibiotics for adults with community-acquired pneumonia. Sensitivity analyses revealed that the initial success rate of each antibiotic was crucial in determining cost-effectiveness. When the number of times antibiotics are taken in a day and the period of therapy were taken into account, AZM was most beneficial with 917,179-1,152,694 yen (US$ 7,643-9,606) per additional QALY over MINO in patients without comorbidity. This result, however, was not applicable to patients with chronic lung disease. MINO was the least expensive and the most cost-effective in empirically treating adult patients with community-acquired pneumonia on an ambulatory basis. AZM provides a higher quality of life for adults without comorbidity with generally acceptable marginal cost. PMID- 11925913 TI - [A case of infected bulla caused by Mycobacterium gordonae]. AB - A 65-year-old man was admitted to Tokyo Metropolitan Geriatric Medical Center because of dyspnea on effort, loss of appetite and general fatigue. Chest radiography on admission showed niveau formation in, and infiltration around, the bulla. Percutaneous drainage of the intrabullous fluid was performed and Mycobacterium gordonae was repeatedly cultured from it. The patient was treated with RFP, INH and CAM and the intrabullous fluid and infiltration around the bulla was markedly improved. Seven cases of pulmonary infection by M. gordonae have been reported in Japan, but this is the first case of bulla infected by this organism. PMID- 11925914 TI - [Etiology, pathogenesis and management of senile inflammatory pulmonary diseases]. AB - We studied the etiology, pathogenesis and management of therapy-resistant inflammatory pulmonary diseases. First, to understand the pathogenesis of rhinovirus (RV) infection-induced exacerbation of bronchial asthma, we infected cultured human tracheal epithelial cells with RV. The epithelial cells produced a variety of proinflammatory cytokines, intercellular adhesion molecules (ICAM-1) and low-density lipoprotein receptor, and increased the permeability across the epithelial cells. These findings suggest that these factors and the increased permeability may cause airway inflammation, resulting in the exacerbation of asthma. Glucocorticoid and bafilomycin inhibited RV infection in the epithelial cells by reducing ICAM-1 expression and RV RNA entry from the acidic endosomes to the cytoplasm. Second, we revealed the mechanisms of aspiration pneumonia induced by silent aspiration in patients with cerebral infarction. We also developed a pharmacologic treatment for preventing aspiration pneumonia with amantadine, which stimulates the dopaminergic neurons; the angiotensin-converting enzyme inhibitors, which decrease substance P catabolism; and cilostazol, which inhibits platelet aggregation and induces cerebral vasodilation. Third, we demonstrated that exhaled carbon monoxide concentrations caused by heme oxygenase-1 upregulation, may be a useful noninvasive means of monitoring airway inflammation and of controlling elderly patients with bronchial asthma. Finally, we demonstrated that microsatellite polymorphism in the heme oxygenase-1 gene promoter is associated with susceptibility to emphysema caused by cigarette smoke in Japanese patients with chronic pulmonary emphysema. PMID- 11925915 TI - [Interstitial pneumonia associated with chronic thyroiditis]. AB - A 56-year-old woman was initially seen in March 1998 with a complaint of back pain. Her thyroid gland was swollen and the level of TSH was elevated at 10 microU/ml. Chronic thyroiditis with hypothyroidism was diagnosed. Negative reactions were obtained for both anti-thyroglobulin and anti-thyroid peroxidase antibodies. Daily treatment with thyroxine 50 mg was started. A cough developed in August 1998, and her chest radiographs revealed multiple bilateral consolidation shadows. A transbronchial lung biopsy specimen demonstrated mononuclear cells infiltration in the alveolar septa with fibrosis, consistent with nonspecific interstitial pneumonia (NSIP). There were no findings clinically or serologically suggestive of collagen-vascular diseases. Ultimately interstitial pneumonia compatible with the NSIP pattern in association with chronic thyroiditis was diagnosed. Oral prednisolone (30 mg/day) and azathioprine (50 mg/day) administration was initiated. The cough and the shadows on the chest radiographs improved. However, one year after completion of the treatment, interstitial pneumonia recurred. Retreatment with prednisolone and azathioprine has resulted in its improvement. PMID- 11925916 TI - [A case of mixed pulmonary infection with Mycobacterium tuberculosis and M. intracellulare due to familial prevalence]. AB - A 76-year-old woman was admitted to our hospital because of an abnormal shadow on chest radiography and a suspected familial prevalence of pulmonary tuberculosis. Her son had previously been admitted to our hospital with pulmonary tuberculosis and tuberculous pleuritis. In the present case, a diagnosis of pulmonary tuberculosis and atypical pulmonary mycobacteriosis was made on the basis of detection of both Mycobacterium tuberculosis and M. intracellulare several times in a sputum culture of anti-fast bacilli. Restriction fragment length polymorphism (RFLP) analysis was then performed for both the woman and her son. The RFLP analysis revealed the same pattern in both mother and son, and confirmed that this was a case of mixed pulmonary infection passed from son to mother. PMID- 11925917 TI - [A case of rebamipide-induced pneumonitis]. AB - A 75-year-old man who complained of progressive dyspnea was admitted to our hospital. He had been treated with rebamipide at an outpatient clinic for the preceding seven months. Atrial blood gas values obtained on admission indicated hypoxemia. Chest radiographs and CT films revealed diffuse interstitial infiltration and minimal pleural effusion. Bronchoalveolar lavage fluid showed an increase in the total cell number, the percentages of eosinophils and lymphocytes, and a decrease in the CD4/CD8 ratio. Transbronchial lung biopsy specimens had thickened alveolar septa with lymphocyte infiltration within the air spaces. After the discontinuation of rebamipide, oral prednisolone was administered, and the patient's symptoms and pulmonary shadows quickly improved. A lymphocyte stimulating test for rebamipide was positive. A challenge test for rebamipide was not performed because informed consent could not be obtained. On the basis of these findings, our diagnosis was rebamipide-induced pneumonitis. PMID- 11925918 TI - [Measurement of anti-type II collagen antibody diagnosis and follow-up useful in a case of relapsing polychondritis]. AB - A 68-year-old man had been suffering from swelling and deformation of both auricles, hoarseness, hearing loss, and a productive cough since November, 1999. Elevation of the inflammatory reaction was noted, together with swelling of the glottis and inflammation of the bronchial mucosa on bronchoscopy. Relapsing polychondritis was diagnosed when an auricle biopsy specimen demonstrated auricle chondritis. The laboratory findings revealed high titers of anti-type II collagen antibody. After steroid treatment, the symptoms improved and the titer of anti type II collagen antibody decreased. The measurement of this antibody was useful for the diagnosis and follow-up of this disease. To prevent the impairment of organs and sudden death, early diagnosis involving the use of anti-type II collagen antibody and the commencement of therapy are important in this disease. PMID- 11925919 TI - [A case of mediastinal teratoma with elevated serum tumor marker levels]. AB - A 27-year-old man complaining of cough was admitted to our hospital because of a giant mediastinal tumor on the chest radiograph. Chest CT and MRI revealed a giant polycystic mediastinal tumor. Chest radiographs on admission showed left pleural effusion due to perforation of the cyst. Laboratory data showed high serum and pleural fluid concentrations of CA 125, CA 19-9, SLX and others. The mediastinal mass was resected and diagnosed pathologically as a mature teratoma. It is reported that patients with mediastinal teratomas often have pleural fluid as a result of self-digestion by pancreatic enzymes, and some mediastinal teratomas have high serum tumor marker levels. We suspected that the high serum tumor marker levels in our case were caused by the high concentrations of tumor markers in the pleural fluid. We suggest that serum tumor marker levels may be useful in the preoperative differential diagnosis of anterior mediastinal cystic tumors. PMID- 11925920 TI - [Successful use of immunosuppressive agents for the treatment of progressive rheumatoid interstitial lung disease: a case report]. AB - Interstitial pneumonia (IP) is sometimes a fatal complication of rheumatoid arthritis (RA). We describe a patient with progressive rheumatoid interstitial pneumonia, who responded to intravenous intermittent cyclophosphamide (IV-CY) and cyclosporine (CsA). A 62-year-old man with rheumatoid arthritis was admitted to this hospital because of dyspnea. Examinations on admission revealed that he had active RA with vasculitis and IP Initially, he responded to high-dose corticosteroid therapy. A lung biopsy performed after initial corticosteroid therapy revealed diffuse interstitial pneumonia with marked infiltrations of macrophages into the air spaces. On corticosteroid therapy with prednisolone 30 mg/day, the IP became exacerbated and was refractory to the current high-dose steroid treatment. He responded to intravenous cyclophosphamide, but his IP remained unstable. After CsA treatment was started, a clinical remission was obtained. In this case, CsA was the most effective agent tried. Clinical and pathological considerations led us to speculate that activated alveolar macrophages played a crucial role in the pathogenesis of steroid-resistant IP in this patient, and that the clinical remission induced by CsA may have been due to its inhibitory effect on alveolar macrophages. PMID- 11925922 TI - [A case of chronic hypersensitivity pneumonia with elevation of serum SP-D and KL 6]. AB - A 73-year-old woman suffering from dyspnea on effort and chronic cough was admitted to our hospital. Chest computed tomography disclosed ground-glass opacities, irregular linear opacities and honeycombing distributed predominantly in the subpleural area. The serum levels of SP-D and KL-6 rose to 889 ng/ml, 1,755 U/ml, respectively. These findings indicated idiopathic pulmonary fibrosis. However, the number of lymphocytes and the CD4/CD8 ratio in the BAL fluid were elevated. Transbronchial lung biopsy specimens demonstrated alveolitis with granuloma formation. The evidence that she had lived in a house with a heavy fungal growth and that tests of precipitation in response to Penicillium corylophilum were positive confirmations of a diagnosis of chronic hypersensitivity pneumonia. PMID- 11925921 TI - [Mycobacterium abscessus infection complicated with diabetes mellitus]. AB - We report a case of Mycobacterium abscessus infection complicated with diabetes mellitus. A 38-year-old man with diabetes mellitus as an underlying disease, was admitted to our hospital because of a productive cough. He had had pulmonary tuberculosis two years before. Chest radiography revealed infiltration in both lung apices and chest CT showed a cavitary lesion in the left upper lobe. Gaffky 2 was found on a sputum smear. However, in the examination of PCR on sputum, not only M. tuberculosis but M. avium complex was negative, and repeated cultures of sputum were positive for M. abscessus. On the basis of the diagnosis of an M. abscessus infection, the patient was initially treated with amikacin, imipenem/cilastatin and levofloxacin during hospitalization while receiving insulin for diabetes mellitus. The smear and culture of sputum became negative for Mycobacterium, and the findings of chest radiography and chest CT improved. After discharge, treatment was continued with clarithromycin and levofloxacin. It is considered that the choice of effective drugs and the additional treatment of an underlying disease are very important for the treatment of a Mycobacterium abscessus infection. PMID- 11925924 TI - [An adult patient with varicella-zoster pneumonia while under inhaled steroid treatment]. AB - Bronchial asthma had been diagnosed in a 33-year-old man, and he had then been treated with a moderate dose of inhaled steroids (fluticason propionate 400 micrograms/day). On year later, he was admitted to our hospital complaining of sore throat, fever, loss of appetite, and skin eruptions. Despite the administration of Acyclovir for three days, varicella pneumonia was diagnosed. Computed tomography of the chest and bronchoscopic examination revealed characteristic findings: nodules with surrounding ground-glass attenuation and multiple vesicles with an ulcerativelesion on the bronchial mucosa, respectively. The demonstration of varicella-zoster virus DNA in a bronchoalveolar lavage specimen by the polymerase chain reaction technique was useful in the formulation of a definitive diagnosis. PMID- 11925923 TI - [An autopsy case of pulmonary and central nervous system metastatic osteosarcoma treated with thirty-six courses of chemotherapy over four years]. AB - A 32-year-old man presented with cough, dyspnea and orthopnea ten years after amputation of the right humerus because of osteosarcoma. Chest radiographs and chest computed tomographs showed left pleural effusion, pericardial effusion and a giant intrathoracic mass, which was histologically diagnosed as a recurrence of the osteosarcoma. After 4 courses of chemotherapy combined with CDDP, the mass in the left upper lobe of the lung decreased in size, and it was then resected. Three months later, new metastatic lesions were detected in the thoracic area. Therefore, 29 additional courses of chemotherapy were administered (36 courses in total over 4 years; including regimens combined with CDDP, carboplatin, high-dose methotrexate, ifosfamide, dacarbazine, vindesine, etoposide, vincristine, taxotere and gemcitabine). In spite of the several courses of chemotherapy, brain and spinal cord metastases appeared, and the patient eventually died of cerebral hemorrhage. During the four years after the first recurrence he had good quality of life as a result of the chemotherapy. PMID- 11925925 TI - [Mechanisms of drug resistance and reversal of the resistance]. AB - The mechanisms for the resistance to anticancer agents have been vigorously studied and many factors that are involved in the resistance were found. Among the members of ABC transporter superfamily, P-glycoprotein, MRP1-5 and BCRP are involved in the drug resistance. LRP, identified as the major vault protein, is also related to drug resistance. PMID- 11925926 TI - [Gene expression profiling of human ovarian epithelial tumors by digo nucleotide microarray]. AB - Gene expression of human ovarian carcinoma cell lines and epithelial ovarian tumors was examined by oligonucleotide microarray for about 6000 human cDNAs. (1) Comparison of gene expression between CDDP-sensitive human ovarian serous adenocarcinoma cell lines and CDDP-resistant cell lines revealed that gamma glutamylcysteine synthetase, glutathione peroxidase-like protein, dehydrogenase (UGDH), NAD(P)H: quinoneoxireductase, glucose-6-phosphatase, ornithine decarboxylase and dihydrodiol dehydrogenase were associated with a mechanism of CDDP-resistance. Comparison of gene expression between taxol-sensitive human ovarian cell lines and taxol-resistant cell lines showed that up-regulation of 30 kinds of gene expression including MDR and semaphorin E in taxol-resistant cell lines. (2) Comparison of gene expression among serous adenocarcinomas, clear cell adenocarcinomas and non-cancerous ovarian tissues by hierarchical clustering demonstrated that clear difference between carcinomas and non-cancerous ovarian tissues but not obvious difference between serous and clear adenocarcinomas. Genes that were up- and down-regulated specifically in these two types of ovarian carcinomas were further selected by the criteria that difference in the mRNA level by more than 4-fold between tumors and non-cancerous tissues. Tissue type specific alterations of gene expression are likely to play important roles in the carcinogenesis of epithelial ovarian tumors. cDNA microarray is a powerful and high-throughput tool to analyze gene expression of cancer development. PMID- 11925927 TI - Analyses by comparative genomic hybridization of genes relating with cisplatin resistance in ovarian cancer. AB - Cisplatin has played a key-role in the management of ovarian cancer patients. Since the mechanisms of cisplatin-resistance have been reported to be multifactorial, it is quite difficult to predict effectiveness of cisplatin-based chemotherapy. In the present study, we have screened abnormal chromosomal regions in cisplatin-resistant and paclitaxel-resistant human ovarian cancer cell lines using comparative genomic hybridization (CGH). Increased copy number at 6q21-25 and decreased copy number at 7q21-36 and 10q12-15 were observed in the cisplatin resistant cell line. Increased copy number at 7q11.2-21 was observed in paclitaxel-resistant cell lines. Messenger RNA of MDR1 located on chromosomal region of 7q11.2-21 was overexpressed in the paclitaxel-resistant cell lines and recognized as a potential mechanism of acquired paclitaxel-resistance. In CGH analyses of 28 primary epithelial ovarian cancer patients, gains of 1q21-22 (p = 0.0183) and 13q12-14 (p = 0.0407) were observed in significantly high abundance in the cisplatin-resistant tumor group, compared with the cisplatin-sensitive tumor group. These genetic alterations were suggested to be potential indicators for drug resistance. PMID- 11925928 TI - New molecular tumor markers for endometrial cancer. AB - Although the International Federation of Gynecology and Obstetrics officially changed the classification system of endometrial cancer from a clinically staged to a surgically staged disease in 1988, optimal management of patients with endometrial cancer is still controversial. Gynecologists happen to experience that patients with tumors that are identical in grade and stage often have significantly different clinical outcomes or responses to therapy. In order to identify an objective biological factor correlating with tumor aggressiveness, many tumor markers have been investigated. So far, CA125 is one of the most reliable tumor marker for adenocarcinoma of the uterus and frequently used in a clinical setting. Recently, with the advent of molecular biological techniques, many genes and regions of the genome related to endometrial cancer have been identified. We undertook a genome-wide screening to detect genetic changes by comparative genomic hybridization (CGH) in primary endometrioid cancers, since CGH analysis provides comprehensive information concerning relative chromosomal losses and gains in tumors by a single hybridization. In this paper, the usefulness of serum tumor markers and the new promising molecular tumor markers for endometrial cancer are discussed. PMID- 11925929 TI - Telomerase activation and MAPK pathways in regenerating hepatocytes. AB - Although there have been many reports on the relationship between the activation of telomerase and carcinogenesis, the role of telomerase in normal cellular growth is still unclear. Recently, the telomerase upregulation during the process of liver regeneration has been reported, but the precise time course of its activity and factors contributing to the activation of telomerase have not yet been fully elucidated. In the present review, we demonstrate the relationship between the activation of the telomerase, the cell cycle progression and the growth-related signaling during the liver regeneration process using an in vivo mouse partial hepatectomy model. Moreover, the importance of the role of the MAPK pathways on the telomerase activity in regenerating hepatocytes is also displayed by using an in vitro culture model. In conclusion, the telomerase activity is upregulated before hepatocytes enter the S phase, and some growth factors such as EGF and HGF contribute to this process. The activation of the growth-related signaling pathways seems to play essential roles in the upregulation of the telomerase activity. PMID- 11925931 TI - Current progress and perspectives in cell therapy for diabetes mellitus. AB - Recent advances of cell transplantation and tissue engineering are remarkable. And also the diabetic treatment using pancreatic beta cells have performed great advances. Even clinical islet transplantation has been considered a common curative treatment for diabetes mellitus in the place of an experimental treatment. Still more the lack of donor's organ as a worst problem of transplantation will be overcome by using the beta cells produced in vitro culture. Therefore diabetes mellitus will be closed to cure in the near future. PMID- 11925930 TI - New approach for the establishment of mouse early embryonic stem cells and induction of their differentiation. AB - Eleven early embryonic stem (EES) cell lines were established using a new novel method. Two cell stage embryos from the ddY mouse strain were cultured in alpha MEM supplemented with 10% fetal calf serum (FCS) and embryotrophic factors (ETFs) and allowed to develop to the trilaminal germ disc embryonic stage. Only small round cells (EES cells) were isolated by the colony isolating technique and subsequently cultured in the same medium containing the ETFs and leukemia inhibitory factors (LIF-10 ng/ml). The newly established embryonic stem (ES) cells isolated from inner cell mass of blastocysts differentiated from two cell stage embryo in culture. The EES and ES cell lines were maintained in an undifferentiated state using Ham's F12 medium supplemented with 10% FCS and 1 ng/ml of LIF. The EES cells maintained their normal genetic and morphological features as well as their potential to differentiate into a broad spectrum of cell types as well as their ability to contribute to all cell lineages in chimeric mice. Moreover, these cell lines changed and differentiated into various kinds of cells by removing LIF and by the addition of ETFs to the vitro culture system. All 11 EES cell lines and 3 ES cell lines formed embryoid bodies; however, cell line EES-4 formed tube-like structures which extended, anastomosed with each other, and finally formed networks when the LIF were absent. Primitive germ organ-like structures composed of 3 germ layers were recognized in the cultures following the administration of ETFs. In conclusion, the new method devised by us is a novel, easy and reliable technique for establishing EES cell lines. PMID- 11925932 TI - Cloned transgenic mouse fetuses from embryonic stem cells. AB - Development of efficient efficient system for genetic modification and large scale cloning of livestock is of importance for agriculture, biotechnology, or human medicine. The mouse, on the other hand, is an ideal model in the basic studies of genetic modification. In this study, we investigated about production of clone mice from established embryonic stem (ES) cell line by nuclear transfer. Further, we had try of production of cloned transgenic mouse fetuses/offspring using ES cells modified with a marker gene, EGFP. With the ES cell line TT2 which is at least 15 passages, reconstructed oocytes developed to 2-8 cell embryos, morulae, or blastocysts (44.8%), and 17.2% of them developed to term (19.5 days post-coitum, dpc). When 40 embryos with the marker gene transferred to 11 surrogate mothers (pseudopregnant females), 5 live fetuses were recognized in the uteli at 13.5 dpc and in these fetuses expression of GFP was observed, but none developed beyond 19.5 dpc. The present results suggest that ES cells can be used tg produce cloned mice. PMID- 11925933 TI - Analysis of gene expression profiles associated with cisplatin resistance in human ovarian cancer cell lines and tissues using cDNA microarray. AB - Gene expression profiles were analyzed by using cDNA microarray for a cisplatin sensitive cell line (KF), and three- and thirty-fold cisplatin-resistant ovarian cancer cell lines (KFr and KFrP200) both showing no p53 mutation within exon 5, 6, 7, 8 and no pglycoprotein overexpression. Expression of GST-pi mRNA increased as the level of resistance to cisplatin became high. Microarray analysis revealed that DNA repair associated genes, i.e., XRCC5, XRCC6, ERCC5, hMLH1 were over expressed in three-fold cisplatin-resistant cell line, KFr as compared to cisplatin-sensitive parental cell line, KF. Apoptosis inhibitors, i.e., IGFR type I and II were over-expressed, and apoptosis inducer, i.e., caspase 3 and BAK were underexpressed in highly cisplatin-resistant cell line, KFrP200 as compared to KFr. As for clinical cases, cDNA microarray was used to compare gene expression profiles directly between two groups, i.e., the chemotherapy (CAP) sensitive group (n = 2) and the resistant group (n = 2). Six genes such as beta tubulin, high-mobility group (nonhistone chromosomal) protein 1, connective tissue growth factor, insulin-like growth factor binding protein 2, alpha tubulin, and RAS related gene were overexpressed in CAP therapy resistance group, whereas seven genes such as CD9 antigen, alpha-2-macroglobulin, caveolin 2, interleukin 1 receptor antagonist, Rho GTPase activating protein 1, reticulon 3, cyclin dependent kinase 10, keratin 7 were underexpressed in CAP therapy resistance group. By increasing clinical case number and gene number of microarray to be used in the analysis of expression profile of gene cluster affecting anticancer drug resistance and sensitivity of the ovarian cancer, it would be possible to apply microarray analysis to personalization of chemotherapy such as selection of effective chemotherapy protocol and prediction of therapeutic effect in the near future. PMID- 11925934 TI - Cloned murine fetuses produced by nuclear transfer using metaphase-arrested embryonic stem cells. AB - We examined whether metaphase nuclei could be used as nuclear donors in nuclear transfer in mice. The reconstructed embryos were developed to fetuses in both the metaphase-nuclear transfer and the G1-nuclear transfer. We also performed enucleation of oocytes following nuclear injection (injection-enucleation method) using microinjection method with a piezo-driven micromanipulator in order to produce the cloned murine fetuses. We found that this method could shorten time for manipulation in comparison with the conventional method performing nuclear injection following enucleation of oocytes (enucleation-injection method). We produced successfully cloned fetuses by the injection-enucleation method. Furthermore, there was no difference of developmental efficiency in reconstructed embryos from between B6D2F1 and ddY strain as oocyte donor. PMID- 11925936 TI - [Okazaki furaguments and discontinuous replication]. PMID- 11925937 TI - [Functional alteration of Bacillus subtilis protease (subtilisin) by engineering of the pro-sequence]. PMID- 11925935 TI - Role of thymidine phosphorylase in Fas-induced apoptosis. AB - Thymidine phosphorylase (TP) has chemotactic and angiogenic activity in vitro, and it promotes tumor growth and inhibits apoptosis in vivo. It plays a key role in the invasiveness and metastasis of TP-expressing solid tumors. KB/TP cells transfected with a TP cDNA have been shown to be resistant to hypoxia-induced apoptosis, suggesting that TP has effects on tumor growth and cell death independent of its effects on angiogenesis. However, the mechanisms of cell death inhibition by TP are unknown. In the present study, we demonstrate that caspase-8 is cleaved in control transfectant KB cells early on during Fas-induced apoptosis. Caspase-8 activation leads to the loss of mitochondrial membrane potential, followed by the release of cytochrome c, the activation of caspase-3, and apoptosis. In contrast, Fas-induced caspase-8 cleavage is inhibited in KB/TP cells, which lead to inhibition of the downstream apoptotic cascade and inhibition of apoptosis. These findings indicate that TP plays an important role in intracellular apoptotic signal transduction in the Fas-induced apoptotic pathway. Therefore, inhibition of TP may suppress the progression of TP overexpressing solid tumors by inducing apoptosis. PMID- 11925940 TI - [The structural biology and infection mechanism of bacteriophage T4]. PMID- 11925938 TI - [Lyme borreliosis, clinical symptoms and terrestrial transmission of etiologic agent, Borrelia burgdorferi]. PMID- 11925939 TI - [Regulation of neurotransmitter release by tyrosine kinases]. PMID- 11925943 TI - [Step-by-step approach to water and electrolyte disorders]. PMID- 11925941 TI - [An actin-regulatory protein, gelsolin functions as a regulator of cell growth and apoptosis]. PMID- 11925942 TI - [Radical-catalyzed enzymatic reactions: structure and mechanism of B12 enzymes]. PMID- 11925944 TI - [Five cases of acute poststreptococcal glomerulonephritis with acute renal failure and nephrotic syndrome]. AB - During the period of 27 years from 1974 to 2000, acute episodes were studied retrospectively in 130 patients under 14 years of age with poststreptococcal acute glomerulonephritis(PSAGN). PSAGN cases have a variable clinical presentation from asymptomatic to severe oliguric acute renal failure(ARF). Proteinuria is nearly always present, but is less than 3.5 g/day. Nephrotic syndrome(NS) is not commonly observed in PSAGN. Among 130 patients, 5 cases had NS with ARF. The present study investigated a spectrum of NS with ARF and the significance of clinical features in PSAGN. PMID- 11925945 TI - [Serum albumin levels and prognosis after commencing chronic dialysis in diabetic patients with end-stage renal failure]. AB - We investigated 1) the association between serum albumin levels(Alb) at the initiation of dialysis and survival after a mean follow-up period of 30.1 +/- 23.1 months following the commencement of chronic dialysis, and 2) the factors associated with hypoalbuminemia at the initiation of dialysis in diabetic patients with end-stage renal failure(ESRF). A hundred and thirty diabetic patients who were initiated on chronic dialysis in our department between January 1992 and November 2000 were studied. Alb and some variables were collected at 12, 6, 3, and 1 month before the initiation of dialysis(-12, -6, -3, -1 M), at the initiation of dialysis(0 M), and 1, 6, and 12 months after commencing chronic dialysis(1, 6, 12 M). Multivariate logistic regression analysis showed that hypoalbuminemia(less than 3 g/d/) at 0 M was significantly associated with an increase in urinary protein excretion per 1 g/day during -12 M and -1 M and total blindness due to diabetic retinopathy(Odds ratio 1.41, 8.83, p = 0.01, 0.03). Cox's proportional-hazard model demonstrated that a decrease in Alb per 1 g/dl at 0 M was significantly associated with survival(Hazard ratio 3.69, p = 0.03, adjusted age, sex, mode of dialysis, and urinary protein excretion during -12 M and -1 M). In addition, a decrease in Alb between -1 M and 0 M was significantly associated with elevated C-reactive protein at 0 M and a decrease in hematocrit between -1 M and 0 M. In conclusion, hypoalbuminemia at the initiation of dialysis, from any cause other than proteinuria, was associated with a poor prognosis after commencing chronic dialysis in diabetic ESRF patients. PMID- 11925946 TI - [An elderly case of ANCA-positive membranous nephropathy]. AB - We report a rare case of nephrotic syndrome in an elderly woman with positive antineutrophil cytoplasmic antibody(ANCA). The patient was 81 years of age and had a history of interstitial pneumonia. She was diagnosed rheumatoid arthritis(RA) at admission. Rapidly progressing renal damage was found with mild microscopic hematuria and positive ANCA. The renal biopsy findings indicated membranous nephropathy. Neither gold nor anti-rheumatic drugs had been previously administered. She may have had an RA-specific membranous nephropathy. Crescentic formation was not clear. With hematuria, the leukocyte infiltration in the capillary lumen and the change in epithelial cells of Bowman's capsules would be histological findings suggesting ANCA-associated nephritis. This is a rare report on membranous nephropathy in an RA patient with ANCA-associated nephritis. PMID- 11925948 TI - [Is viscocanalostomy really all it's made out to be?]. PMID- 11925947 TI - [A case of allopurinol-induced muscular damage in a chronic renal failure patient]. AB - A 73-year-old woman with chronic renal failure developed generalized muscular weakness and pain 6 days after the start of allopurinol treatment(200 mg/day). Routine laboratory tests revealed elevated levels of serum creatine kinase, and the patient was clinically diagnosed as rhabdomyolysis, due probably to severe myositis. A high level of serum oxipurinol, the chief active metabolite of allopurinol, was also revealed. The muscular weakness was relieved in seven weeks with intermittent hemodiafiltration. PMID- 11925949 TI - [Effects of various eye drops on corneal wound healing after superficial keratectomy in rabbits]. AB - PURPOSE: The effects of various eye drops on corneal wound healing, particularly in the subepithelial haze area, investigated histologically following superficial keratectomy in rabbits. MATERIAL AND METHODS: Mechanical superficial keratectomy was performed in rabbit eyes. Tranilast, betamethasone, hyaluronic acid and diclofenac eye drops were administered after the procedure. Physiological saline was used as a control. Corneas were excised 1, 2, 3, and 4 weeks after keratectomy, labeled with 3H-thymidine or 3H-proline, and subjected to autoradiography. RESULTS: In the control and diclofenac groups, corneal haze occurred three weeks after keratectomy. Histological examination revealed accumulation of proliferating keratocytes and active synthesis of collagen in the subepithelial area. In the tranilast and betamethasone groups, formation of corneal haze was reduced compared to the controls. The proliferation of keratocytes and the production of collagen in the corneal stroma were inhibited by these drugs. In the hyaluronic acid groups, corneal haze was decreased. In this group, although the proliferation of keratocytes was activated compared to the controls, abnormal accumulation of keratocytes in the subepithelial area was not detected. CONCLUSION: Tranilast and betamethasone decrease the formation of subepithelial haze by inhibiting keratocyte proliferation and synthesis of extracellular matrix in the corneal stroma. Hyaluronic acid, on the other hand, inhibits subepithelial haze by promoting physiologic wound healing. PMID- 11925950 TI - [Effect of tonometry on a glaucoma population study]. AB - PURPOSE: The purpose of this study was to evaluate the influence of tonometry in detecting the occurrence of glaucoma. METHODS: The subjects, 845 out of 3,488 residents aged 40 years or older, were examined according to standard protocols, including tonometry, slit lamp examination, fundus photography, and automatic perimetry as a recall examination. The intraocular pressure in each subject was measured by both Goldmann applanation tonometer(GAT) and noncontact tonometer CT 70 (NCT). RESULTS: The mean +/- standard deviation intraocular pressure measured by GAT was 15.52 +/- 2.57 mmHg, and 15.03 +/- 2.90 mmHg by NCT. There was a statistically significant correlation(p < 0.0001). The difference between pairs of measurements by GAT and NCT was 0.50 +/- 1.93 mmHg. There was no influence of tonometry in detecting the incidence of glaucoma, which was 4.14%; primary open angle glaucoma 0.59%, normal tension glaucoma 2.6%, primary angle-closure glaucoma 0.47%, and other types of glaucoma 0.48%. The detection of ocular hypertension was different, and was 2.13% with GAT and 2.72% with NCT. CONCLUSION: In our study, the influence of tonometry in detecting the incidence of glaucoma was very small. A noncontact tonometer is considered to be useful for glaucoma population study. PMID- 11925951 TI - [Accuracy of suture adjustment in sliding noose type adjustable strabismus surgery]. AB - PURPOSE: To clarify the efficacy of adjustable suture strabismus surgery with sliding noose, we evaluated the accuracy of suture adjustment. METHODS AND SUBJECTS: Thirty-four patients with various types of strabismus participated [age range: 12-79 years, range of far deviation: 4-123 prism diopters(PD)]. Under sub Tenon anesthesia, a recession(with or without an additional resection or a muscle transposition) was performed with an adjustable suture(Guyton's procedure), and the suture was adjusted 6-24 hours after the surgery. Based on the initial postoperative examination performed 1-4 weeks(mean: 2.8 weeks) after the surgery, errors in the far deviation from the individual target were analyzed. RESULTS: Twenty-six(76%) patients required suture adjustment 1-8 times. In 50% and 75% of our patients, the errors were within +/- 0.8 and +/- 2.0 PD, respectively, whereas 2(6%) patients with esotropia showed an undercorrection larger than 10 PD. Distribution of the errors was the same throughout the range of pre-operative deviation. CONCLUSION: By using the sliding noose type adjustable suture, a pin point alignment of the eyes can be established in more than 50% of cases, at least, in the early post-operative period. PMID- 11925952 TI - [Analysis with magnetic resonance imaging of lesions in cerebral achromatopsia]. AB - PURPOSE: To localize the lesions associated with cerebral achromatopsia. METHODS: We examined 20 patients with homonymous hemianopsia caused by cerebral infarction(17 men and 3 women aged 49 to 81 years; mean age, 65.1 years). Ishihara plates, standard pseudoisochromatic plates(part 2) and the panel D-15 test were used to examine color perception. Color matching tasks and color naming tasks were used to test color recognition. We tried to apply functional magnetic resonance imaging(fMRI) to lesion analysis in the brain. RESULTS: Cerebral achromatopsia was diagnosed in four patients. The analysis showed that lesions in the infracalcarine area(Brodmann's area 18 and 19) were associated with cerebral achromatopsia. Additionally, the lesions associated with failure of the panel D 15(PD-15) test were located more anterior than the lesions associated with failure of Ishihara plates. CONCLUSION: We show evidence that lesions in the anterio-ventral temporo-occipital area are associated with cerebral achromatopsia. This result is in accord with past observations(autopsy, fMRI and positron emission tomography). PMID- 11925953 TI - [Retinal thickness and changes with age]. AB - PURPOSE: We studied the morphological changes occurring with aging in the macula using optical coherence tomography(OCT). METHODS: Forty-seven eyes from 47 normal volunteers were studied. The subjects ranged in age from 21 to 79 years and their refractive errors were within +/- 3.00 diopters. The measurement of the retinal thickness was done by OCT. The retinal thickness was evaluated at five points: the foveola, and 1 mm superior, inferior, nasal, and temporal to the foveola. The axial length of the eye and the refraction were also measured in all subjects. RESULTS: The retinal thickness was 142 +/- 15 microns at the foveola, 257 +/- 18 microns at the superior point, 255 +/- 18 microns at the inferior point, 246 +/- 20 microns at the temporal point, and 261 +/- 21 microns at the nasal point (mean +/- standard deviation). All points except the foveola showed reduced retinal thickness with age. Thus, attenuation of the retinal thickness in the parafoveal area was correlated significantly with age. However, no change in the retinal thickness at the foveola was observed with increasing age. Neither axial length nor refractive error biased this result. CONCLUSION: Our results demonstrated that the macular thickness changes with aging, but no obvious change could be detected in the foveola. PMID- 11925954 TI - [Outcome of surgery for bilateral third nerve disorder]. AB - PURPOSE: To review the outcome of surgery for bilateral third nerve palsy. CASES AND METHODS: The series comprised 16 cases. The eye deviation in the primary position averaged -27.0 degrees horizontally. Surgery was aimed at bringing both eyes into alignment in the primary position by recession-resection of the horizontal muscles. Transposition of the superior oblique was performed for complete third nerve palsy. Surgery was initially performed on the nonfixating eye. The fellow eye received further surgery for residual disorders. RESULTS: Within 6 months after surgery, the eye deviation in the primary position averaged -0.7 degrees horizontally. After longer follow-up, the eye deviation averaged 4.7 degrees horizontally. Postoperatively, diplopia in the primary position was absent in 11 and present in 5 cases. CONCLUSION: Surgery for bilateral third nerve paresis or palsy induced lasting cosmetic or functional improvements in the majority of cases. PMID- 11925956 TI - [A case of secondary glaucoma studied by optical coherence tomography of the anterior ocular segment]. AB - PURPOSE: To compare slit lamp biomicroscopy or gonioscopy with optical coherence tomography (OCT) and to assess the efficacy of OCT in a case of anterior segment disease. CASE: A 74-year-old male who had bilateral keratoconus. The left eye was aphakic, and a penetrating keratoplasty was performed on it, as well as Nd:YAG laser capsulotomy. The prognosis was good in the early postoperative stage. But 6 months postoperatively, we could not control the intraocular pressure and judged that a second operation might be needed. Before the operation, we tried to get images of the anterior segment of this eye using slit lamp biomicroscopy, gonioscopy, and OCT. RESULTS: Findings obtained by OCT were more useful than those obtained by slit lamp biomicroscopy or gonioscopy to determine the method of operation. DISCUSSION: This case substantiates the view that observation of the anterior ocular segment by OCT is useful for such cases, because in cases of corneal disease we can not get much information about the deep and endothelial side of the cornea from slit lamp biomicroscopy. PMID- 11925955 TI - [Outcome of combined phacoemulsification, intraocular lens implantation and viscocanalostomy]. AB - PURPOSE: To assess the pressure-lowering effect, visual acuity, and intra- and postoperative complications of combined cataract surgery, intraocular lens(IOL) implantation and viscocanalostomy. METHODS: Combined viscocanalostomy, phacoemulsification, and IOL implantation was performed on 45 eyes, which were followed for more than 6 months after surgery. The average preoperative intraocular pressure(IOP) was 20.2 +/- 3.1(mean +/- standard deviation) mmHg, and the average medication number was 1.7. A prospective study of these cases was performed. RESULTS: IOP was significantly reduced after surgery, being 13.6 mmHg at 3 months and 15.1 mmHg at 6 months postoperatively. The visual acuity was improved more than 2 lines in 38 eyes. Disruption of Descemet's membrane was noted in 5 eyes and microperforation in 9 eyes as intraoperative complications. Postoperative complications were not vision threatening and included short term hyphema in 34 eyes, fibrin deposition in 5 eyes and IOP spike(> 5 mmHg) in 5 eyes. There were no cases of hypotony, choroidal detachment, or endophthalmitis. CONCLUSION: Combined viscocanalostomy, cataract extraction, and IOL implantation was a safe and efficacious way to reduce IOP. PMID- 11925958 TI - [Famotidine induced parkinsonism in a case of essential tremor]. PMID- 11925957 TI - [Malabsorption syndrome due to chronic pancreatitis resulting in severe zinc deficiency]. PMID- 11925959 TI - [Progressive multifocal leukoencephalopathy with chronic renal failure primarily involving the cerebellum: an autopsy case]. PMID- 11925961 TI - [Familial papillary thyroid carcinoma]. PMID- 11925960 TI - [Ruptured aortic dissection into the left atrium (DeBakey II)]. PMID- 11925962 TI - [Pheochromocytoma presenting as acute heart failure]. PMID- 11925963 TI - [Secondary amyloidosis complicated with membranous nephropathy]. PMID- 11925964 TI - [Recurrent Fisher syndrome]. PMID- 11925965 TI - [Chronic hepatitis C with HTLV-I associated myelopathy (HAM) treated with interferon]. PMID- 11925966 TI - [Follicle-stimulating hormone producing pituitary adenoma with hyponatremia]. PMID- 11925967 TI - Treatment of degenerative lumbar spinal stenosis. PMID- 11925968 TI - Prevention of venous thromboembolism after injury. PMID- 11925969 TI - An evaluation of beta-blockers, calcium antagonists, nitrates, and alternative therapies for stable angina. PMID- 11925970 TI - Diagnosis and treatment of acute bacterial rhinosinusitis. PMID- 11925971 TI - Diagnosis and treatment of swallowing disorders (dysphagia) in acute-care stroke patients. PMID- 11925972 TI - Evaluation of cervical cytology. PMID- 11925975 TI - Drawing a spark from darkness: John Walsh and electric fish. AB - John Walsh's research on electric fish, carried out between 1772 and 1775, proved fundamental for demonstrating that electricity might be involved in animal physiology, and, moreover, in favouring a period of great progress in both the physiology and physics of electrical phenomena. However, Walsh is hardly known to modern neuroscientists and is largely neglected by science historians also. One of the reasons for this neglect is that he never published his 'crucial experiment', that is the production of a spark from a discharge of the electric eel. PMID- 11925977 TI - A.F.W. Schimper: from the 'oecology of plant distribution' to the 'functional ecology of terrestrial ecosystems'. PMID- 11925979 TI - Designing nature reserves: adapting ecology to real-world problems. AB - Establishing nature reserves is the foundation of regional conservation strategies. The science of nature reserve design is relatively new, having emerged alongside the interdisciplinary field of conservation biology over the past three or four decades. Its goal is to apply scientific ideas and methods to the selection and design of nature reserves and to related problems, such as deciding what kinds of buffer zones should surround reserves or how to establish corridors to link reserves and allow organisms to move from one area to another. As in other areas of conservation biology, designing nature reserves is a 'crisis' science, whose practitioners are driven by an acute sense of urgency over the need to stem the loss of species caused by human population growth. PMID- 11925980 TI - Royal Society of Tropical Medicine and Hygiene Meeting at Manson House, London, 18th January 2001. Pathogen genomes and human health. Mycobacterial genomics. AB - The small size of their genomes made bacterial ideal model organisms for the emerging field of genomics. Elucidating the genome sequences of mycobacteria was particularly attractive owing to the difficulties inherent in their manipulation. The slow growth rate, clumping, and requirement for category III containment make manipulation of Mycobacterium tuberculosis-complex strains laborious. M. leprae presents even greater problems as it has resisted all attempts at axenic culture. Availability of genome sequence data promised to accelerate our knowledge of the fundamental biology of these organisms, and to offer clues to the basis for their virulence, tropism and persistence in the host. This article will focus on what the genome sequences of M. tuberculosis and M. leprae have taught us about these pathogens, and how comparative genomics has exposed some of the fundamental differences between the species. PMID- 11925981 TI - Copulation songs in three siblings of Lutzomyia longipalpis (Diptera: Psychodidae). AB - We present the results of recording male courtship songs of the sandfly Lutzomyia longipalpis. The striking differences in the songs from 3 Brazilian populations of this sandfly with 3 distinct male pheromones support the 3 sibling species previously proposed based on this characteristic. PMID- 11925982 TI - A polyvinyl alcohol (PVA)-containing medium for permanent mounting of helminth eggs. AB - A method involving use of a polyvinyl alcohol (PVA)-containing medium and a single coverslip is described for permanent mounting of helminth eggs. This simple method, which avoids some disadvantages of double-coverslip methods, can be applied for mounting different helminth eggs, especially in teaching laboratories. PMID- 11925983 TI - Efficacy of praziquantel against Schistosoma mansoni in northern Senegal. PMID- 11925984 TI - Risk factors associated with asymptomatic infection by Leishmania chagasi in north-east Brazil. AB - Various factors have been associated with a predisposition to the development of clinical American visceral leishmaniasis (AVL). However, little information is available about the factors that predispose to asymptomatic infection. To identify the risk factors associated with asymptomatic infection, a study was carried out between July 1997 and June 1998 on children aged 0-5 years in the districts of Vila Nova and Bom Viver in the municipality of Raposa in the island of Sao Luis, State of Maranhao, Brazil. A questionnaire containing socioeconomic, demographic and epidemiological data was used. The delayed-type hypersensitivity (DTH) test was carried out on 639 children in the first phase, and on 572 in the second, 7 months after the first survey, using Leishmania amazonensis antigen. Infection was determined by enzyme-linked immunosorbent assay (ELISA) in 638 children during the first phase, and in 572 during the second. Six outcome measures were used: initial prevalence, final prevalence and incidence, each determined by DTH and ELISA. The incidence of infection was 10.8% when determined by DTH and 28.5% when determined by ELISA. After adjustment for confounding variables using Cox regression, infection by L. chagasi was associated with child's age (> or = 2 years), location of the dwellings (Vila Nova) and reporting of relatives with AVL. Bathing outside the house and playing outdoors between 18:00 and 20:00 were identified as risk factors in some analyses but not in others. Presence of intra- and peridomestic Lutzomyia sandflies and animals such as dogs or chickens in the house or in the neighbourhood appeared as risk factors in some analyses but in others they unexpectedly seemed to protect from infection. Malnutrition was not found to be associated with infection. PMID- 11925985 TI - Patterns of helminth infection and relationship to BCG vaccination in Karonga District, northern Malawi. AB - Surveys of enteric and urinary helminth infections were carried out in 1999 among 501 schoolchildren and among 320 adolescents and young adults participating in a study of immune responses to BCG vaccine in Karonga District, northern Malawi. Hookworm, Schistosoma mansoni and S. haematobium infections were detected in 64%, 27% and 20% of schoolchildren and in 55%, 40% and 25% of the immunology study subjects, respectively. Other helminths were appreciably less common. The prevalence of 'at least one' helminth infection was 76% among schoolchildren, ranging from 60% to 92% in the 4 schools, and was 79% in the immunology study participants. There was no evidence for an association between the presence of a BCG scar and presence or intensity of infection with worms in the schoolchildren, nor evidence that BCG vaccination of adolescents and young adults had any effect on the prevalence of helminth infections 1 year later. PMID- 11925986 TI - High death rates in health care workers and teachers in Malawi. AB - High death rates are reported in health care workers (HCWs) and teachers in urban areas of Malawi. The present study was carried out to determine the annual death rate in HCWs and primary school teachers working in semi-urban and rural areas of Malawi, and to try to ascertain the main causes of death. Forty district and mission hospitals in Malawi were visited. A record was made of the number of clinical and nursing-based HCWs in each hospital in 1999, the number of deaths in that calendar year and reported causes of death. A record was also made of the number of teachers working in 4 primary schools nearest to each hospital in 1999, the number of deaths in that calendar year and reported causes of death. There were 2979 HCWs, of whom 60 (2.0%) died. There were 4367 teachers of whom 101 (2.3%) died. Annual death rates, calculated per 100,000 people, were significantly higher in male HCWs compared with female HCWs (2495 versus 1770, RR 1.17, 95% CI 1.14-1.20, P < 0.001), and significantly higher in female teachers compared with male teachers (2521 versus 1934, RR 1.14, 95% CI 1.11-1.17, P < 0.001). In male HCWs and teachers the highest death rates were in those aged 35 44 years. In female HCWs and teachers, the highest death rates were in those aged 25-34 years and 35-44 years, respectively. Reported causes of death in HCWs were tuberculosis (TB) in 47%, chronic illness in 45% and acute illness in the remainder, while in teachers the causes were TB in 27%, chronic illness in 49% and acute illness in 25%. Chronic illness, thought to be due to AIDS, and TB were the common causes of death. The current high death rates from AIDS and TB will have a crippling toll on the health and education sectors, and effective ways of reducing these death rates must be found. PMID- 11925987 TI - Mosquito nets for the elderly? AB - Nine-year follow-up (ending 1999) of survival of 3738 individuals in a malaria endemic area of Papua New Guinea found that the use of mosquito nets was associated with a large reduction in mortality in people aged > or = 40 years as well as in children aged < 5 years. There may be substantial benefits of malaria transmission reduction for older people, that have been overlooked in public health programmes and burden of disease calculations. PMID- 11925988 TI - Characteristics of a cholera outbreak, patterns of Vibrio cholerae and antibiotic susceptibility testing in rural Malawi. AB - The cumulative cholera attack rate in an epidemic in Malawi in 1999/2000 was 59/100,000 population, case-fatality rate 4%, and 98% of all cases presenting to health facilities required intravenous therapy. Microbiological studies showed high resistance of Vibrio cholerae to commonly recommended antibiotics, predominant Ogawa serotypes and no O139 isolates. PMID- 11925990 TI - Isoenzymatic identification of Leishmania isolates from repeated clinical human leishmaniasis episodes in Catalonia (Spain). AB - Forty human strains of Leishmania infantum isolated in 1985-99 from 17 patients with repeated cutaneous, mucosal or visceral leishmaniasis episodes in Catalonia (Spain) were examined by isoenzyme electrophoresis. Six zymodemes were revealed: MON-1, MON-24, MON-28, MON-29, MON-33 and MON-34. In 2 patients 2 different zymodemes were identified in consecutive episodes. PMID- 11925989 TI - Prevalence and intensity of Wuchereria bancrofti antigenaemia in Sri Lanka by Og4C3 ELISA using filter paper-absorbed whole blood. AB - In Sri Lanka 2741 people from Matara, an endemic area for Wuchereria bancrofti, were examined in 1996/97 for microfilariae by 60-microL blood smear and for circulating filarial antigens by Og4C3 ELISA using filter paper-absorbed whole blood. The overall prevalence of microfilaraemia was 3.4%, and that of antigenaemia 14.4%. The prevalence of antigen-positive and microfilaria-negative people was 11.3%. Analysed by age-group, antigenaemia prevalence was similar in all groups, and the average number of antigen units was already very high in the age-group < 10 years, indicating that the infection started in early childhood. Among those who were antigen positive, the microfilaria prevalence was lower in females than in males. Diethylcarbamazine treatment eliminated microfilariae in 78% of the positives. However, 17 months after the treatment, antigenaemia was still positive in 76% of those who were parasitologically cured. PMID- 11925991 TI - Incidence of Trypanosoma cruzi infection in two Guatemalan communities. AB - The prevalence of human infection by Trypanosoma cruzi was assessed using an enzyme-linked immunosorbent assay (ELISA) in a serological survey in 1998 of 2 rural communities (SMH and PS) in Guatemala. In SMH (Department of Zacapa), where Rhodnius prolixus was the principal vector, the seroprevalence amongst 373 people tested was 38.8%. In PS (Department of Santa Rosa), where the main vector was Triatoma dimidiata, 8.9% of the 428 people tested were seropositive. The overall prevalence of seropositivity was higher in females than in males in both SMH (40% vs 36%) and PS (11.9% vs 4.9%), although this difference was significant only in PS. Historical seroconversion rates, estimated retrospectively by fitting a transmission model to the age-prevalence curves, were 3.8% per year in SMH and 0.5% per year in PS. There was some indication of a recent reduction in incidence in both villages. In PS, but not in SMH, both the observed prevalence and the estimated incidence rates were significantly higher in females than in males. PMID- 11925992 TI - Genetic variability within Trypanosoma brucei gambiense: evidence for the circulation of different genotypes in human African trypanosomiasis patients in Cote d'Ivoire. AB - For 23 Ivoirian patients infected by Trypanosoma-brucei gambiense, isolation and genetic characterization using PCR and microsatellite primers were performed (in 1996-99) using 2 different isolates (A and B) from each patient. When using TBDAC 1/2, 7 genotypes were observed, and DNAs A and B for 2 patients were different. This might be the first evidence of the presence of 2 different genotypes of T. b. gambiense group 1 in the same patient. PMID- 11925993 TI - Search for Enterocytozoon bieneusi infection in wild monkeys in Cameroon. AB - Faecal samples collected from 42 wild monkeys in Cameroon were examined for microsporidia by light microscopy (using Weber trichrome and Uvitex 2B stains) and by PCR (using Enterocytozoon bieneusi specific primers). None of the 42 samples was positive, suggesting that wild monkeys do not represent a major reservoir for microsporidia in Central Africa. PMID- 11925994 TI - Sandfly fauna of endemic leishmaniasis foci in Anzoategui State, Venezuela. AB - A census of the sandfly fauna was undertaken in 1993-98 in 5 endemic leishmaniasis foci situated at different altitudes in Anzoategui State, Venezuela. From the 17 species of Lutzomyia identified, we believe that Lu. ovallesi, Lu. panamensis and Lu. gomezi are the probable vectors of cutaneous leishmaniasis, while Lu. evansi might transmit visceral leishmaniasis. PMID- 11925995 TI - Evaluation of the ICT whole-blood antigen card test to detect infection due to Wuchereria bancrofti in Sri Lanka. AB - The sensitivity, specificity and cost effectiveness of an immunochromatographic card test (ICT, AMRAD) for the diagnosis of bancroftian filariasis were estimated against 2 standard parasitological techniques: thick blood film (TBF) and Nuclepore membrane filtration (NMF). Individuals were selected from endemic localities in the Western Province (n = 213) and from the non-endemic Central Province (n = 29) of Sri Lanka. Blood was collected between 21:00 and midnight. Sixty microlitre of non-heparinized blood, and 1 mL and 100 microL of heparinized blood were used in TBF, NMF and ICT, respectively. NMF was positive in 31.5% (67/213) of the endemic group, with a mean microfilaria (mf) count of 343/mL (range 8-1782, SD 422). All 67 were positive by ICT (sensitivity 100%), but only 63 by TBF (sensitivity 94%). Among the endemic population there were 12 who were mf negative but antigen positive by ICT. There were, however, no false positives among the non-endemic controls, indicating the possibility that the ICT may in fact be more sensitive and 100% specific. Thus, ICT filariasis test appears to be more effective (both sensitive and specific) than TBF or NMF in diagnosing infection in lymphatic filariasis. The direct unit recurrent costs of the 2 survey tools, TBF and ICT, were US$ 0.30 (Rs. 27/=) and US$ 2.75 (Rs. 248/=), respectively. The high cost of the ICT may be offset by other factors that are difficult to cost. PMID- 11925996 TI - The increase in occurrence of typhoid fever in Cameroon: overdiagnosis due to misuse of the Widal test? AB - This study investigates the cause of an apparent increase in occurrence of typhoid fever in Cameroon. The reasons explored include an overdiagnosis of the illness related to poor performance of the Widal test in laboratories and interpretation by prescribers. Questionnaires were used in 1996 to evaluate the use and interpretation of the Widal test, and checklists were used to assess its laboratory performance in 2 of the 10 provinces in Cameroon. The majority of prescribers from 20 health facilities (an average of 76% of the doctors and 61% of the nurses) could detect patients who truly had positive Widal tests and needed treatment. However, an average of 48% of the doctors and 84% of the nurses would treat patients who did not require treatment based on the Widal test result. Patients may therefore be treated unnecessarily. Most (88%) of the visited laboratories performed the Widal rapid slide agglutination test as opposed to the conventional tube agglutination test. About 14% of the laboratories that performed the rapid slide agglutination test had a score above average for each criterion evaluated. Misdiagnosis of typhoid fever leads to unnecessary expenditure and exposure of patients to the side-effects of antibiotics. In addition, misdiagnosis may result in delayed diagnosis and treatment of malaria, and other acute febrile illness. PMID- 11925998 TI - The Brugia malayi genome project: expressed sequence tags and gene discovery. AB - To advance and facilitate molecular studies of Brugia malayi, one of the causative agents of human lymphatic filariasis, an expressed sequence tag (EST) based gene discovery programme has been carried out. Over 22,000 ESTs have been produced and deposited in the public databases by a consortium of laboratories from endemic and non-endemic countries. The ESTs have been analysed using custom informatic tools to reveal patterns of individual gene expression that may point to potential targets for future research on anti-filarial drugs and vaccines. Many genes first discovered as ESTs are now being analysed by researchers for immunodiagnostic, vaccine and drug target potential. Building on the success of the B. malayi EST programme, significant EST datasets are being generated for a number of other major parasites of humans and domesticated animals, and model parasitic species. PMID- 11925997 TI - The distribution of anti-Salmonella antibodies in the sera of blood donors in Yaounde, Cameroon. AB - This cross-sectional study in late 1996 on a group of 230 consecutive blood donors in Yaounde, Cameroon, found a baseline titre of 1:100 for antisalmonella O antibodies. The seroprevalence and baseline titre for antisalmonella H antibodies was much higher (1:400), suggesting that the H antibody is of limited diagnostic value in Cameroon. PMID- 11925999 TI - Cryptic Plasmodium falciparum parasites in clinical P. vivax blood samples from Thailand. AB - Polymerase chain reaction detection revealed cryptic Plasmodium falciparum infections in 21 of 160 samples collected from Thai patients diagnosed (by microscopy) with vivax malaria. The clinical and biological significance of these mixed infections is discussed in the context of chloroquine resistance and the low inoculation rates which characterize malaria epidemiology in Thailand. PMID- 11926000 TI - Cerebral melioidosis in Singapore: a review of five cases. AB - A variety of neurological manifestations of infection with Burkholderia pseudomallei have been described including cerebral abscesses, which are a well recognized form of neurological melioidosis. The optimal antibiotic therapy for this condition has not been defined; however, combinations of intravenous antibiotics are frequently used in the early stages. Five cases of melioidosis involving brain abscesses are described which presented in Singapore over the past 3 years (1997-2000), 4 of which cases had evidence of disseminated infection. Despite profound neurological deficits and low Glasgow Coma Scale scores at presentation in 3 of these cases, all survived after surgical drainage and prolonged courses of intravenous ceftazidime or imipenem, and only 2 of whom had residual neurological impairment. One incidental finding on computed tomography (CT) or magnetic resonance imaging (MRI) scans not described before as an association with cerebral melioidosis was sinusitis in 4 out of the 5 cases. PMID- 11926001 TI - Leishmania (Viannia) spp. dissemination and tissue tropism in naturally infected dogs (Canis familiaris). AB - First evidence is presented for Leishmania (Viannia) spp. dissemination and tissue tropism in the domestic dog. Using PCR and histology, parasites were detected in the conjunctiva, lung, lymph nodes and ovaries of 2 naturally infected Peruvian dogs. The detection of parasites in the blood indicates that parasite dissemination to those organs may have been haematogenous. PMID- 11926002 TI - Profile of snakebite envenoming in western Maharashtra, India. AB - Ninety-one cases of snakebite were admitted between January 1998 and January 2001 to the general hospital at Mahad, 180 km south of Mumbai in western Maharashtra, India. Twenty-nine (31.9%) patients, when admitted to hospital, brought the snakes responsible for the bites (20 kraits, 9 Echis carinatus). Forty-five (49.5%) patients had snakebite without envenoming; 27 patients had local fang marks without local and systemic manifestations; 18, reporting 24 h after the bite, had local oedema. Twenty-six (28.6%) patients were paralyzed. Twenty-five cases who reported between midnight and 08:00 showed no local signs of envenoming. One patient, who reported at 16:00, had local oedema with blebs and a history of cobra bite. Ten (11.0%) patients died: 3 on the way to hospital and 7 during treatment. Out of 7 cases transferred for tertiary care, 6 recovered and 1 died on the way to Mumbai. Early administration of anti-snake venom (ASV), endotracheal intubation and timely intervention with manual ventilation by Ambu bag and anticholinesterase treatment for Elapidae (krait and cobra) envenoming are crucial for saving lives. Clinical confirmation of snakebite with envenoming was by identification of the dead snake brought by victims and by clinical signs and symptoms such as paralysis (krait and cobra) and rapid progressive local oedema with active bleeding (E. carinatus and Russell's viper). The 20-min whole blood clotting test is a simple, rapid and reliable test of coagulopathy. It is also of use in assessing the effectiveness of ASV and is of crucial importance for controlling administration of expensive ASV in relation to restoration of blood coagulability. PMID- 11926003 TI - An economic comparison of chloroquine and sulfadoxine-pyrimethamine as first-line treatment for malaria in South Africa: development of a model for estimating recurrent direct costs. AB - The relative cost-effectiveness of chloroquine (CQ) and sulfadoxine-pyrimethamine (SP) as first-line antimalarial therapy in southern Africa is of great interest to policymakers, clinicians and researchers in the subregion. A model was developed to access the cost-effectiveness of replacing CQ with SP as first-line treatment in Mpumalanga, South Africa, where malaria is seasonal and the population is non-immune. In-vivo drug resistance levels were used to derive a 'resistance variable' for each drug, which was used to compare the costs to the public healthcare provider associated with either therapeutic option. Costs including drugs, staff time, transport, maintenance, utilities, training and consumables were determined and subjected to Monte Carlo simulation and subsequent analysis to generate an average cost-effectiveness ratio (ACER) with confidence intervals for each drug. SP was found to be 4.8 (95% CI 3.3-6.7) times more cost-effective than CQ in Mpumalanga at 1997 resistance levels and costs, despite the far greater cost per treatment course of SP (US$ 4.02 as opposed to US$ 0.22 for CQ) in South Africa. At the price of SP in Kenya and Uganda (US$ 0.47-4.80 per treatment course), the ACER for SP does not change materially, increasing to between 5.1 and 5.6. Resistance emerged as the factor that most influenced the ACER of a specific drug. Indirect costs, compliance, changes in effectiveness and costs over time and costs of adverse events were not included in the model owing to paucity of data and logistical difficulties. Since most of these are likely to be similar in both drug models, the relative ACER is unlikely to be significantly altered by their inclusion. PMID- 11926004 TI - The Lys-76-Thr mutation in PfCRT and chloroquine resistance in Plasmodium falciparum isolates from Uganda. AB - Recent molecular studies of chloroquine (CQ) resistance of Plasmodium falciparum have demonstrated an association between a mutation in the PfCRT gene and CQ resistance. We identified wild type and mutant alleles of the PfCRT codon 76 in baseline pre-CQ treatment P. falciparum isolates collected during 1999 and investigated their relationship to CQ efficacy in 3 different sites with different levels of CQ parasite resistance in Uganda. Of 32 isolates from Mulago Hospital, all were mutant (100%), while of 45 isolates from Tororo, 5 (11%) were mixed wild type and mutant and 40 (89%) were mutants only. Of 41 isolates from Apac, 13 (32%) were mixed wild type and mutant whereas 28 (68%) were mutants only. The finding of 100% prevalence of the Thr-76 mutant allele in all isolates at the 3 sites was remarkable. We found no association between the presence of Thr-76 mutation and treatment outcome at all the sites. However, the prevalence of the wild-type Lys-76 allele was higher in Apac, an area with lower CQ parasite resistance, compared to Tororo and Mulago which have relatively higher CQ parasite resistance. The Thr-76 allele as a marker of CQ resistance is probably useful in regions where the allele frequency has not yet plateaued. PMID- 11926005 TI - Do mutations in Plasmodium falciparum dihydropteroate synthase and dihydrofolate reductase confer resistance to sulfadoxine-pyrimethamine in Iran? AB - The key codons of dihydropteroate synthase (dhps) and dihydrofolate reductase (dhfr) genes implicated in sulfadoxine-pyrimethamine (SP) resistance were determined by allele-specific polymerase chain reaction in Plasmodium falciparum isolates collected in 1999 from 35 Iranian patients treated with SP. Seven isolates had Glu-540 dhps allele but 5 of these isolates were characterized to possess wild-type dhfr alleles. Seven additional isolates were polyclonal with mixed Lys- and Glu-540. The key dhfr mutation associated with pyrimethamine resistance, Asn-108, was found in 4 isolates. In one patient the presence of Lys- and Glu-540 in dhps and Asn-108 and Arg-59 in dhfr was associated with treatment failure. However, more studies are needed to determine whether clinical response to SP and mutations in these genes are correlated in Iran. PMID- 11926006 TI - Assessment of the neurotoxicity of oral dihydroartemisinin in mice. AB - High doses of the oil-soluble antimalarial artemisinin derivatives artemether and arteether, given by intramuscular injection to experimental mammals, produce an unusual pattern of selective damage to brainstem centres predominantly involved in auditory processing and vestibular reflexes. We have shown recently, in adult Swiss albino mice, that constant exposure either from depot intramuscular injection of oil-based artemisinin derivatives, or constant oral intake carries relatively greater neurotoxic potential than other methods of drug administration. Using the same model, oral dihydroartemisinin suspended in water was administered once or twice daily at different doses ranging from 50 to 300 mg/kg/day for 28 days. The neurotoxic potential of the oral dihydroartemisinin was assessed and compared to that of oral artemether and artesunate. Oral artemether, artesunate, and dihydroartemisinin had similar neurotoxic effects with no significant clinical or neuropathological evidence of toxicity at doses below 200 mg/kg/day. These data indicate that once and twice daily oral administration of artemether, artesunate and dihydroartemisinin is relatively safe when compared to intramuscular administration of the oil-based compounds. PMID- 11926007 TI - Pulmonary hemodynamic and tidal volume changes during exercise in heart failure. AB - BACKGROUND: Impairment of lung mechanics, increase of pulmonary artery pressure and limitation of exercise capacity are common findings in chronic heart failure. The objective of the present study was to evaluate whether pulmonary mechanics are correlated with pulmonary hemodynamics, whether both are correlated with the functional capacity and whether the time course of their changes during exercise correlates with the exercise capacity. METHODS: We performed a cardiopulmonary exercise test (breath by breath analysis of ventilation and gas exchange, cycloergometer, 25 W increments every 3 min) with pulmonary hemodynamic monitoring in 38 heart failure patients. The parameters were analyzed at rest, 1 min after the work rate increase and at peak exercise. RESULTS: A significant linear correlation with peak oxygen consumption was found at rest for: mean pulmonary artery pressure (mPAP, r = -0.56), right atrial pressure (RAP, r = 0.42), pulmonary wedge pressure (PWP, r = -0.53), and total pulmonary (TPR, r = 0.53) and pulmonary vascular resistances (PVR, r = -0.45); after 1 min of exercise for: cardiac index (CI, r = 0.49), mPAP (r = -0.57), RAP (r = -0.60), PWP (r = -0.45), and TPR (r = -0.67) and PVR (r = -0.38); at peak exercise for: tidal volume (r = 0.63), CI (r = 0.63), RAP (r = -0.43), TPR (r = -0.65) and PVR (r = -0.43). A significant linear correlation with peak oxygen consumption was found, for the increment between rest and 1 min of exercise, for RAP (r = -0.58) and CI (r = 0.42) and, for the increments between rest and peak exercise, for tidal volume (r = 0.79) and CI (r = 0.61) and, for the ratio between the increment between rest and 1 min of exercise/increment between rest and peak exercise, for mPAP (r = -0.42), RAP (r = 0.51) and CI (r = -0.54). The same ratio of increment of mPAP (r = 0.39) and CI (r = 0.36) correlated with that of tidal volume. CONCLUSIONS: This study provides evidence of a strong correlation between the respiratory function and pulmonary vascular pressure changes during exercise in heart failure. PMID- 11926008 TI - Prognostic value of 6-minute walk corridor testing in women with mild to moderate heart failure. AB - BACKGROUND: The prognostic usefulness of 6-min walk testing (6 MWT) was reported both in patients with severe as well as in those with moderate systolic heart failure. However, more than 80% of the patients investigated were males and the value of the test had never been separately evaluated in women. The aim of the present study was to prospectively assess the prognostic value of 6 MWT in women with mild to moderate congestive heart failure. METHODS: Ninety-five women with congestive heart failure (mean age 63.9 years) were enrolled in the study. All patients were followed for a mean period of 38 months in order to assess the event-free survival (death, heart transplantation). The prognostic value of the distance walked during 6 MWT was compared to the NYHA functional classification and to the echocardiographically-derived indexes of heart function. The results were then compared with those of a group of 129 male patients with similar clinical characteristics. RESULTS: Thirty-four female patients (36%) died of cardiovascular causes and 2 underwent heart transplantation. In the same period, the overall survival in male patients was 74% and 3 patients were successfully transplanted. Regardless of gender, the 3-year event-free survival was significantly lower in patients who walked < 300 m during 6 MWT compared to that observed in those with an intermediate (300-400 m) or high level (> 400 m) of performance. At univariate and multivariate analyses, the NYHA class and the distance covered during 6 MWT were significantly related to survival in both genders while with regard to the left ventricular ejection fraction, this relationship was observed only in women. The patient's age was not related to the mortality. CONCLUSIONS: A distance < 300 m during 6 MWT identifies a subgroup at high risk of death even in female patients with mild to moderate heart failure. PMID- 11926009 TI - The risk functions incorporated in Riscard 2002: a software for the prediction of cardiovascular risk in the general population based on Italian data. AB - BACKGROUND: The purpose of this analysis was to produce risk functions for the prediction of cardiovascular diseases based on Italian epidemiological data and suitable for the use in a PC program dedicated to the estimate of risk. METHODS: Three studies were used for the purpose: the Italian Rural Areas of the Seven Countries Study, the Gubbio Population Study and the ECCIS study, for a total of 9771 men and women aged 35 to 74 years and followed for a period lasting 5 to 6 years. The risk factors used for the prediction of cardiovascular events were sex, age, body mass index (derived from height and weight), mean blood pressure (derived from systolic and diastolic blood pressures), non-HDL cholesterol (derived from total and HDL cholesterol), HDL cholesterol, diabetes (yes-no), heart rate, and daily cigarette consumption. The endpoints were the first major coronary event, the first major cerebrovascular event, and the first major cardiovascular event (either one between the previous two plus major peripheral artery diseases). The model employed for the analysis was the accelerated failure time model. RESULTS: Having excluded those already presenting with a cardiovascular disease and those with missing values, a total of 9089 subjects were included in the models. In a period lasting 5 or 6 years, a total of 211 coronary, 64 cerebrovascular and 269 cardiovascular events occurred and were considered for analysis. Coefficients from the coronary model suggested a significant association of all risk factors except body mass index and diabetes (marginal significance). Coefficients from the cerebrovascular model suggested a significant association limited to age and mean blood pressure. Coefficients from the cardiovascular model suggested a significant association of all risk factors except body mass index. The discrimination between cases and non-cases was satisfactory with proportions of 37.0, 52.3 and 37.8% of observed cases in decile 10 of the distribution of the estimated risk for the three endpoints respectively. CONCLUSIONS: The three models were used as a mathematical core for the construction of a PC software for the prediction of major cardiovascular events in Italy. PMID- 11926010 TI - Transcatheter closure of the patent ductus arteriosus with new-generation devices: comparative data and follow-up results. AB - BACKGROUND: Many devices currently used for the closure of the patent ductus arteriosus are claimed to be safe and cost-effective, but only few data exist with respect to the gold standard of the Rashkind occluder. The aim of this study was to assess the efficacy and safety of three new different devices and to compare the results to those of a control group of patients carrying a Rashkind occluder. This should provide the basis for further cost-analysis studies. METHODS: The records of all patients who underwent closure of the patent ductus at our Institution from April 1989 to May 2001 were reviewed. Eighty patients (median age 10.3 years, median weight 27.6 kg) were treated (25 with a Rashkind device, 11 with Duct-Occlud coils, 35 with Cook detachable coils, 9 with the Amplatzer system). RESULTS: Kaplan-Meyer estimates of long-term complete occlusion of the ductus showed, compared to the Rashkind device, a significant improvement for the Cook and Amplatzer (p = 0.025 and p = 0.003, respectively) devices but not for the Duct-Occlud coils (p = 0.165). One patient of the Duct Occlud group (9%) and 3 with the Rashkind device (12%) featured a significant residual shunt and needed a second intervention. The complication rate was 4% for the Rashkind occluder, 5% for the Cook coils, 9% for the Duct-Occlud system, and 11% for the Amplatzer device (p > 0.05). CONCLUSIONS: The new devices are as safe as the Rashkind occluder and provide effective treatment. The Cook coils and the Amplatzer occluder offer better results compared to the Rashkind and Duct-Occlud devices. PMID- 11926011 TI - Successful treatment of anteroseptal accessory pathways by transvenous cryomapping and cryoablation. AB - Radiofrequency is the most commonly used energy source for the treatment of cardiac arrhythmias. Despite its high success rate, radiofrequency energy may sometimes present limitations, especially in case of anteroseptal atrioventricular accessory pathways. In these patients, inadvertent atrioventricular block may occur during or after the procedure and a high recurrence rate of conduction over the accessory pathway is observed. Since the late 1970s, cryosurgery has been an integral part of the management of cardiac arrhythmias, and recently, animal and clinical studies demonstrated the feasibility and safety of applying percutaneous catheter cryoablation technology. These studies also showed that reversible cryomapping of high-risk arrhythmogenic sites can be performed before creating permanent lesions. In this preliminary report, we describe the successful use of percutaneous cryoablation for the permanent interruption of conduction over anteroseptal accessory pathways. PMID- 11926012 TI - Coronary ostial stenosis after aortic valve replacement. AB - Coronary ostial stenosis is a life-threatening complication of aortic valve replacement. We describe the case of a patient who developed symptoms and signs related to coronary insufficiency 4 months after aortic valve replacement. In view of a coronary angiogram revealing a 99% left main stem stenosis, an urgent revascularization procedure was performed. Intraoperative inspection revealed a dense fibrous reaction of the aortic root involving the left coronary ostium. The postoperative course was uneventful and since the operation the patient remained asymptomatic. The possible mechanisms involved in the pathogenesis of coronary ostial stenosis after aortic valve replacement and the surgical strategy to be employed for the treatment of this complication are discussed. PMID- 11926014 TI - Images in cardiovascular medicine. Pacemaker failure caused by hyperkalemia. PMID- 11926013 TI - Cardiovascular glycoside-like intoxication following ingestion of Thevetia nereifolia/peruviana seeds: a case report. AB - Some plants contain glycoside compounds which determine cardiovascular symptoms similar to those observed after acute toxic digoxin administration. The present case report involves a patient who showed important cardiovascular symptoms following the ingestion of Thevetia nereifolia/peruviana seeds. About 30 min after ingestion, a 65-year-old man presented with dizziness, giddiness, numbness and a burning sensation, diarrhea, sweating, vomiting and ECG changes. At the time of admission he presented with tremors; his body temperature was 37 degrees C, and blood analysis gave the following results: K 5.6 mEq/l, myoglobin 176 IU, troponin T 0.10 ng/ml, PO2 69 mmHg, PCO2 37.4 mmHg, pH 7.33, HCO3- 19.9 mEq/l, hemoglobin 14.8 g/dl, saturation 92.5%. Echocardiography showed a left ventricle with normal global and segmentary contractility. The following days, the patient showed a reduction, until total resolution, of the atrioventricular block and of the alterations of the ST segment. The ectopic beats also resolved; K value before discharge was 4.4 mEq/l. On the third day, the serum levels of digoxin were 0.15 ng/ml. This case report is important because it describes all the cardiovascular and non-cardiovascular signs of glycoside toxicity in an adult patient who accidentally swallowed only two seeds (non-fatal dose) of Thevetia. PMID- 11926015 TI - Images in cardiovascular medicine. "Corkscrew" aortic arch branching pattern. PMID- 11926016 TI - Magnetocardiography: current status and perspectives. Part I: Physical principles and instrumentation. AB - Magnetocardiography (MCG) is a non-invasive and risk-free technique allowing body surface recording of the magnetic fields generated by the electrical activity of the heart. The MCG recording system allows spatially and temporally accurate measurements of the very weak magnetic fields produced by currents flowing within myocardial fibers during cardiac activity. MCG has now been around for over 30 years, but only recently has progress in instrumentation put the technique on the verge of clinical applicability. This review summarizes the physical principles, instrumentation, main clinical applications and perspectives for the clinical use of MCG. This first part is devoted to the description of the physical principles and instrumentation. PMID- 11926017 TI - Current standard of care in patients affected by coronary heart disease in Italy: the MC'95 study. AB - BACKGROUND: The aims of MC'95 were to verify i) whether the main risk factors for coronary heart disease had been recorded in the hospital medical records of patients admitted for coronary events; ii) how modifiable risk factors had been managed by drug therapy and lifestyle advices following hospitalization; iii) whether first-degree blood relatives of patients had been screened for coronary heart disease risk factors. METHODS: The survey concerned 40 hospital units across Italy admitting patients with acute coronary events or offering interventional cardiology or cardiac surgery. Two thousand and four hospital records (50 for each unit) of consecutive patients (< or = 74 years, 21% women) discharged within 6 months before initiation of the study (November 1995) were reviewed and the patients were invited for a follow-up examination from 6 to 9 months after hospital discharge. RESULTS: The prevalence of patients positive for risk factors reported was: 58.5% for high total cholesterol (> or = 5.2 mmol/l), 46.9% for high blood pressure (> or = 140/90 mmHg), 35.2% for smoking, 20.2% for high blood glucose (> 7 mmol/l), and 14.2% for obesity (body mass index > or = 30 kg/m2). A number of medical records did not provide complete information on risk factors: 24.1% for body mass index, 11.0% for serum cholesterol, and 11.6% for a family history of premature coronary heart disease. Eighty-nine percent of the patients attended the follow-up examination. Their care during the follow-up period was managed by general practitioners (30.5%), cardiologists (37.2%) and/or at hospital outpatient clinics (42.4%). At follow-up examination the prevalence of hypercholesterolemia increased to 62.2%, that of uncontrolled hypertension decreased to 32.2%, and the percentage of smokers decreased to 13.6%. The adherence to the drug treatment prescribed at discharge was as follows: 69.6% for lipid-lowering agents, 81.2% for ACE-inhibitors and beta-blockers, and 90.8% for antiplatelet agents. A family history of premature heart disease was reported in 55.1% of patients but screening for coronary heart disease risk factors was extended to relatives in less than 25% of cases. CONCLUSIONS: The prevalence of modifiable risk factors is high in patients with different clinical manifestations of coronary heart disease. The prescriptions for secondary prevention and their efficacy after hospital discharge and the surveying of relatives for primary prevention need to be improved. For this purpose, a more coordinated intervention of primary care physicians, cardiologists and cardiac surgeons is mandatory. PMID- 11926018 TI - A population-based study on overt heart failure in Lombardy (survey of hospitalization in 1996 and 1997). AB - BACKGROUND: The aim of this study was to get updated information about the incidence and outcome of heart failure hospital admission in the whole population of the largest Italy region. METHODS: The Lombardy regional database of hospital records (Information System and Quality Control Bureau) provided the requested information: all the heart failure cases admitted to all the institutions of the region were selected by the ICD9 code of congestive heart failure (428-) as the principal or secondary diagnosis. The available data included: diagnosis and surgical procedures (up to four), date of birth, dates of admission and discharge, outcome, DRG code, discharge unit code, gender. The data so obtained concern the hospital discharges in 1996 and 1997. RESULTS: 32,093 cases were selected. The mean ages were 74.1 +/- 11.5 years for males and 80.6 +/- 10.7 for females in 1996, and 71.8 +/- 11.7 for males and 78.3 +/- 10.6 for females in 1997. Most of the cases were classified as appertaining to DRG 127--pure heart failure (56.7%). Heart failure associated with myocardial infarction accounted for 7.3% of cases. The remaining cases (36.0%) were classified as appertaining to heterogeneous medical and surgical DRGs. In 1997, the in-hospital mortality was 14.7% for patients aged > 80 years; 9.7% for patients aged between 71 and 80 years; 7.4% for patients aged between 61 and 70 years and 6.9% for patients < 61 years of age with the exclusion of pediatric cases. The in-hospital mortality was different between discharge units: 2.8% of in-patients discharged from cardiology units, and 10.3% of in-patients discharged from general medicine units. The readmission rate at 1 month was 5.49% whereas that within 1 year was 14.3%. CONCLUSIONS: Heart failure mortality differences between hospital units are not explained by age and by comorbidity. The readmission rate was lower than in previous reports. PMID- 11926019 TI - [More risks and fewer treatments: the paradox about elderly patients with acute coronary syndromes]. AB - The majority of patients with acute coronary syndromes are elderly subjects. They are at a high risk of events; in fact, they usually arrive to hospital late and this delay nullifies the advantages of reperfusion; they often present with a large and complicated acute myocardial infarction and the short-term mortality for such patients is 3-5 times higher than that observed for younger subjects. Although they are a high-risk population, paradoxically they receive thrombolytic therapy, beta-blocker drugs and acetyl salicylic acid less frequently than younger patients and they are rarely submitted to interventional procedures. In this overview, we analyze the reasons of this paradox and we suggest some management guidelines. The risk of bleeding associated with thrombolytic drugs is the main reason justifying the limited use of reperfusion therapy in elderly patients. The identification, in each patient, of the risk factors for bleeding permits stratification of such patients into different classes of risk. This may be of help to the physician in distinguishing those patients who are candidates for thrombolytic therapy from those who are not, reserving for the latter other therapeutic strategies such as primary coronary angioplasty. In elderly patients with unstable angina or myocardial infarction, a careful and early risk stratification should serve as a guide when establishing the indication for interventional procedures. The latter should be encouraged in those patients in whom the risk of bleeding is high and whose overall clinical picture does not contraindicate such a therapeutic strategy. In such cases, invasive therapy can really improve the patient's quality of life. PMID- 11926020 TI - [The elderly patient with heart failure at hospital cardiology units]. AB - The majority of patients hospitalized for heart failure are elderly. In order to point out the clinical characteristics and the quality of care of elderly heart failure patients we evaluated available data from national databases and observational studies. Elderly patients have more severe clinical manifestations, multiple etiologies and comorbid diseases, frequent hospital admissions and a worse prognosis. As many as 30-50% of elderly patients with heart failure may have normal systolic function. In the elderly, evidence-based treatments are relatively underused with often inappropriately low doses. However, this "underuse" largely depends on the higher "frailty" of these patients (i.e. multiple coexisting diseases, disability, socio-environmental factors) and the lack of definite evidence on efficacy and safety of treatments in the very elderly. Thus, there are opportunities to improve quality in many aspects of care, such as instrumental and multidimensional assessment and out-of-hospital management. Targeted clinical trials and rigorous observational studies to favor the implementation of specific guidelines into clinical practice in the elderly are needed. PMID- 11926021 TI - [Transposition of great arteries. Understanding its pathogenesis]. AB - Transposition of the great arteries (TGA) is a frequent and severe cardiac defect. In patients with this malformation, diagnostic and surgical results and the long-term prognosis significantly improved in the last years. From the embryological point of view there are two main theories: 1) the anomalous infundibular rotation, and 2) the anomaly of the aortico-pulmonary septum. Both of them still present important limits. Moreover, TGA is difficult to reproduce by animal experiments, but interesting data, using retinoid acid in pregnant rats, are nowadays available, as well as there are interesting data from the epidemiologic studies on human teratologic agents. TGA is rarely associated with genetic syndromes and with additional extracardiac anomalies. A few cases are in relation with DiGeorge syndrome with deletion of chromosome 22q11. On the contrary TGA is significantly prevalent, in association with other cardiac and extracardiac anomalies, in children with lateralization defects, heterotaxy and asplenia syndrome (right isomerism). However in patients with heterotaxy and polysplenia syndrome (left isomerism) TGA is significantly more rare. In mice with mutation of Smad2 and NODAL, two genes involved in the lateralization process, some cases of TGA, with or without right isomerism of the lungs, were reported. Moreover, in families with heterotaxy some cases with congenitally corrected TGA were reported and a new gene associated with heterotaxy, CRYPTIC, can present mutations in patients with "isolated" TGA. A recent study on familiar recurrence of TGA shows in the same family some cases of TGA and of corrected TGA so that a monogenic inheritance (autosomic dominant or recessive) with variable phenotypic expression can be suggested. The normal righthand spiralization of the heart is genetically determined in cases of situs solitus and d-loop of the ventricles. This pattern is not present in cases of TGA presenting a parallel position of the great arteries. On the basis of these observations and according to new epidemiologic and genetic data some cases of TGA should be classified in the group of the anomalies of lateralization and ventricular loop. The mystery is still present but perhaps some gleams of light are appearing. PMID- 11926022 TI - [Tissue Doppler in the assessment of myocardial function in stress echocardiography]. AB - Recently, stress echocardiography has emerged as a valuable tool for the diagnosis and evaluation of coronary artery disease, but its interpretation still remains subjective, relying on image quality and reader's experience. These problems could be overcome by quantitative analysis of wall motion. Tissue Doppler provides quantitative information on regional myocardial systolic and diastolic velocities that can be displayed either in spectral mode or color coded, reflecting the peak velocity increment induced by exercise or dobutamine administration. Pulsed wave tissue Doppler allows to measure regional instantaneous myocardial velocities with high temporal resolution and has been shown valuable for detecting stress-induced changes of both myocardial systolic and diastolic function. This method may also identify myocardial viability by measuring increase in systolic peak velocity at low-dose of dobutamine in dysfunctional myocardial segments. Color coded tissue Doppler resolves mean velocities with higher spatial resolution, and post-processing analysis of digital acquired images has been shown feasible and reproducible. Myocardial velocity gradient is a more sensitive parameter compared to the simple measurement of the peak endocardial systolic velocity for evaluating myocardial ischemia during dobutamine echocardiography. From the raw data, it is also possible to measure strain and strain rate. These new parameters have the potential to differentiate between wall motion and contractility, with obvious implications when applied to stress echocardiography. In conclusion, tissue Doppler is able to quantify regional myocardial function. After a large scale validation, this technique will be incorporated with stress echocardiography in clinical practice. PMID- 11926023 TI - [T-wave alternation: a new method for identifying patients at risk of malignant ventricular tachyarrhythmia]. AB - T wave alternans (TWA) is a change, in the microvolt range, of T wave amplitude on ABABAB sequence. TWA depends on heart rate, being optimally analyzed at a target frequency of 110 b/min. Initial studies used atrial pacing to reach the target frequency and reported a sensitivity and specificity of 89% for TWA in predicting tachyarrhythmic events. Subsequently, similar results were obtained using ergometric test to reach the target frequency, a less invasive and more "physiologic" approach to increase heart rate. This method became therefore the elective system to increase heart rate in order to evaluate the presence of TWA by means of spectral analysis. At present, various groups of high arrhythmic risk patients have been evaluated, including those with a recent myocardial infarction, congestive heart failure, implantable cardiac defibrillator and clinical indication to programmed ventricular stimulation. In all clinical conditions analyzed, TWA analysis demonstrated a good diagnostic accuracy, suggesting a possible clinical use of the test in these settings. PMID- 11926024 TI - [The process of drug development. The "case" of spironolactone]. AB - In the progressive understanding of the heart failure syndrome different models have followed. Current pathophysiology model suggests that heart failure is a neurohormonal disorder: an abnormality in cardiac function and a disturbance in myocardial contractile force is the initiating event in heart failure, and the clinical expression and progression of the disease occurs as a result of subsequent extracardiac alterations. Treatment with drugs capable of antagonizing the effects of neuroendocrine activation, such as angiotensin-converting enzyme inhibitors, beta-blockers and antialdosterone agents have induced a significant reduction in morbidity and mortality in heart failure patients. The progressive understanding of the circulating homeostasis mechanisms moved from the systemic to the paracrine aspects of the neurohormonal systems and contributed to the development of new drugs. Moreover, the analysis of a drug's development can be an important step in the medical knowledge advancement. In this sense, aldosterone and spironolactone became an interesting "case", which is analyzed in this paper, going through the steps that have promoted the spironolactone from a weak diuretic to a neurohormonal drug and, finally, to a "central" drug. PMID- 11926025 TI - [Role of clopidogrel in acute coronary syndromes without ST-segment elevation]. AB - Unstable angina and no ST-segment elevation myocardial infarction are acute coronary syndromes (ACS) typically characterized by minimal or little myocardial damage, but often heralding "classical" infarction or coronary death in the short or medium term. Coronary thrombi, mainly composed by platelets, are the major pathogenetic component of ACS and an effective therapeutic target. The hemostatic balance can be shifted towards an antithrombotic state in the clinical setting. Although the thrombogenic stimuli acting in each individual patient, their intensity as well as the prothrombotic factors that may contribute to an inappropriate thrombus growth, remain largely unknown. To this purpose the modulation of platelet aggregation is a particularly effective and practical therapeutic target. The availability of a new antiplatelet agent, clopidogrel, with a proven efficacy in the prevention of cardiovascular events, suggests the opportunity of reviewing the mechanisms of instability and the therapeutic strategies in patients with ACS. In this review we analyze: 1) the composition of coronary thrombi in patients with ACS, 2) the mechanisms of thrombus development and growth, 3) the mechanisms of platelet adhesion and aggregation, 4) the characteristics of clopidogrel, 5) the efficacy of the various therapeutic strategies, 6) the possible implications of the adjunctive therapy with clopidogrel for the treatment and prevention of cardiovascular events in ACS with no ST-segment elevation. PMID- 11926026 TI - [The Torino Network Project. Global management of acute myocardial infarction from the field to the hospital]. AB - A unidirectional clinical pathway for acute myocardial infarction from out-of hospital setting to the coronary care unit and catheterization laboratory could lead to mortality reduction. The ongoing "Progetto Torino Network. Gestione globale dell'infarto miocardico acuto prime ore dal territorio all'ospedale" is based on this statement and described in the three-structural, diagnostic therapeutical, multimedial issues. This project represents the historical evolution of our involvement in out-of-hospital cardiac emergency management. PMID- 11926027 TI - [Myocardial infarction redefined based on the consensus document of the ESC/ACC]. AB - An expert committee of the European Society of Cardiology and the American College of Cardiology has recently proposed new and more precise criteria for the diagnosis of myocardial infarction, entailing both relevant implications in clinical practice and scientific, epidemiological and organizational aspects. The Board of the Emergency Area of the National Association of Hospital Cardiologists (ANMCO) will review the document and analyze the issues of major concern. PMID- 11926028 TI - [Heart rupture in acute myocardial infarction: multicenter observational study of the coronary unit of Piedmont]. AB - BACKGROUND: The aim of this study was to prospectively evaluate the incidence of cardiac rupture during myocardial infarction (MI) as well as the predictive value of the main cardiac rupture risk factors. METHODS: The study was carried out in 17 coronary care units (CCU) between January and December 1999 in the Piedmont region (Italy). RESULTS: The incidence of cardiac rupture was 1.4% of the total number of MI (n = 3041). Data from 13 out of 17 CCU showed the following causes of death during MI: 66% heart failure, 16% cardiac rupture, 7% arrhythmias, 11% others. Twenty-seven percent out of 44 cardiac ruptures had prior angina, 9% prior MI; 24% of patients were diabetic; 38% had anterior wall MI; 62% infero postero-lateral MI; 86% showed ST-segment elevation, and 79.5% developed Q waves. Thrombolysis was administered in 39% of cases. Forty-three percent cardiac ruptures occurred within 24 hours. Electromechanical dissociation was present in 73% of cases, syncope and hypotension in 43%, bradycardia in 30%. An echocardiogram was performed in 89% of cases in the suspicion of cardiac rupture but only 45% showed severe pericardial effusion. One patient was referred to surgery but he died in the postoperative period. Autoptical diagnosis was made in 32% of cases. All patients died. The analysis of some qualitative variables (gender, thrombolysis, MI localization, ST-segment/non-ST-segment elevation) in 8 out of 17 CCU, between the cardiac rupture group (n = 22) and the MI group (n = 1330) showed a significant result only for the female gender. CONCLUSIONS: Cardiac rupture is the second cause of death during MI after heart failure; there is a higher incidence of cardiac rupture in infero-postero-lateral MI, after the first 24 hours particularly in the female gender; there is a low global incidence (1.4%). PMID- 11926029 TI - [Heart rupture during myocardial infarction: the long-lasting challenge for the hospital cardiologist is close to the last battle]. AB - Cardiac rupture is a catastrophic and generally unexpected (although not always unpredictable) complication of acute myocardial infarction, and still represents a challenge for the hospital cardiologist. In fact, diagnostic criteria are not available which allow us to detect in the individual patients an impending rupture. On the other hand the early diagnosis of the rupture, although possible if based on clinical and echocardiographic criteria, allows only a small number of the patients to survive, and cannot be considered as a satisfying solution for this clinical problem. Starting from these discouraging considerations, the finding is underscored that heart rupture only rarely occurs among patients treated with primary coronary angioplasty. Thus primary angioplasty (unlike systemic thrombolysis) seems to be able to prevent the postinfarction cardiac rupture. As a consequence, primary angioplasty should be considered instead systemic thrombolysis for the reperfusion treatment of patients with acute myocardial infarction and clinically at risk for cardiac rupture (i.e., those aged > 60 years and with ST-segment elevation at hospital admission). PMID- 11926030 TI - [Spontaneous dissection of 2 coronary branches in a young man. Report of a clinical case]. AB - Spontaneous coronary dissection is responsible for acute coronary syndromes particularly in females during and in the peri-partum period. It rarely occurs in patients without atherosclerotic coronary plaques. We report a particular clinical course of a 39-year-old patient with spontaneous dissection of two coronary arteries. His clinical course suggested only medical treatment, with aspirin, beta-blockers and ACE-inhibitors. At 3 months of follow-up the patient is free of symptoms. PMID- 11926031 TI - [Right heart failure in a patient with giant mediastinal dermoid cyst]. AB - This case report describes the management of a rare giant mediastinal dermoid cyst causing right heart compression and failure. The precise relationships with vital structures were preoperatively defined at transesophageal echocardiography and computed thoracic tomography. These allowed us to predict the likely diagnosis and therefore plan a limited surgical approach. A cardiopulmonary perfusion team was on stand-by and continuous intraoperative transesophageal echocardiography was used to document the restoration of hemodynamic parameters. PMID- 11926033 TI - [Two cases of false troponin I increase in patients with heterophile antibodies]. AB - Cardiac troponin T and I are highly sensitive and specific biochemical markers for the detection of myocardial damage and they are now considered the preferred markers for the diagnosis of myocardial infarction. Despite this, in some cases elevations in the serum levels of cardiac troponin T and I are not associated with a final diagnosis of cardiac necrosis. These false-positive results are to be related to different interferences in immunometric assays. We report 2 cases of false-positive troponin I results due to heterophilic antibodies. Two women admitted to the Emergency Department with acute chest pain persistently showed, in serial blood samples, elevated and constant values of troponin I serum levels. The results were confirmed as being false positives by treatment of the samples with heterophilic blocking reagent (Scantibodies Laboratory, Santee, CA, USA). Coronary artery disease was excluded at coronary angiography and at stress testing in the first case and at stress myocardial perfusion imaging in the second case. In clinical practice, in case of persistently elevated but constant values of cardiac troponin without the time interval of release characteristic of acute syndromes, it is important to bear in mind the possible occurrence of false positive cardiac troponin results due to the presence of heterophilic antibodies. PMID- 11926032 TI - [Concomitant left atrial myxoma and atrial fibrillation in patients with peripheral embolism]. AB - In the present article, 2 cases of left atrial myxoma and peripheral embolism in patients with concomitant paroxysmal or permanent atrial fibrillation are reported. In both cases, the embolic event was considered to be related to atrial fibrillation and the diagnosis of atrial myxoma was established later on at echocardiography. In view of the above, transesophageal echocardiography should be always performed after an embolic event, even in patients with established atrial fibrillation. PMID- 11926034 TI - [Cardiogenic shock caused by acute occlusion of mammary artery bypass: successful treatment with primary angioplasty in the immediate postoperative period]. AB - The early occlusion of a graft in the postoperative phase is not infrequent and is associated with high morbidity and mortality. A non-surgical solution, which is quick and effective, is an important achievement. A 65-year-old woman underwent myocardial revascularization. Three grafts were performed: left mammary artery to the left anterior descending coronary artery, saphenous vein to the right coronary artery and to the marginal branch. One hour after surgery, the patient developed progressive arterial hypotension which rapidly progressed to cardiogenic shock. The electrocardiogram showed anterior and lateral lesions. The angiogram revealed the occlusion of the first third of the left mammary artery. An angioplasty was performed in this vessel and in the left anterior descending coronary artery. The final angiogram showed good patency of the mammary artery and of the left anterior descending coronary artery. The hemodynamic conditions of the patient improved rapidly, with good pressure values. Primary angioplasty proved to be feasible in the early postoperative phase with good results; it could be a valid alternative to reoperation with lower morbidity and mortality. PMID- 11926040 TI - [Cardiovascular risk or coronary risk?]. PMID- 11926041 TI - Use of a hospital-based accident and emergency unit by children (0-12 years) in Alkharj, Saudi Arabia. AB - This cross-sectional study was carried out in order to define the extent and nature of inappropriate use of the accident and emergency (ER) unit in our community. Data were collected for all children aged < or = 12 years who attended the ER from 1 January to 31 March 1999. A total of 3329 children (2010 boys and 1339 girls) attended the ER during the study period. Respiratory tract infection was the most common presenting complaint (33.5%), followed by trauma (15.3%), bronchial asthma (13.7%) and gastro-intestinal problems (6.6%). Using the triage criteria of Alkharj Military Hospital, most of the boys (57%) and 32.6% of the girls were considered to be simple, primary care cases. The rate of referral to specialist clinics was 9.5% and only 4.7% were actually admitted. Attendance at night (53.5% boys, 59.8% girls) was significantly greater (p < 0.00001) than during the morning (0700-1500) and afternoon shifts (1500-2300 hours). This study shows that a large number of children attend the ER with non-urgent complaints. PMID- 11926042 TI - Kangaroo care and multiple births. PMID- 11926043 TI - Plasma exchange in a child with systemic lupus erythematosus antiphospholipid antibodies and profound deafness. PMID- 11926044 TI - Risk factors and cumulative incidence of anaemia among human immunodeficiency virus-infected children in Uganda. AB - Anaemia has not been well characterised among HIV-infected children in sub Saharan Africa. Baseline prevalence and cumulative incidence of anaemia (haemoglobin < 110 g/L) were 91.7% and 100% and, for moderate anaemia (haemoglobin < 90 g/L), were 35.1% and 58.4%, respectively, among 225 HIV infected children followed from 9 to 36 months of age. Hospitalisation, suspected tuberculosis, malaria and height-for-age Z-score <-2 were significantly associated with moderate anaemia. Moderate anaemia and weight-for-height Z-score <-2 were associated with mortality. Anaemia is common and associated with increased mortality in HIV-infected children. PMID- 11926045 TI - Growth and micronutrient disturbances in stable, HIV-infected children in Cape Town. AB - This prospective study of 60 stable, HIV-infected children in an economically deprived setting was designed to document anthropometric and micronutrient disturbances. Investigations included CD4+ counts, anthropometry and plasma levels of albumin, transthyretin, retinol-binding protein (RBP), vitamins A, B6, E and B12, and folate, zinc and copper. The median age was 25 months. Thirty-two per cent had mild, 48% moderate and 20% severe clinical features, and 80% were moderately or severely immunosuppressed. Twenty-eight per cent had a weight Z score <-2.0 and 58% a height Z-score <-2.0. Many children had micronutrient deficiencies: albumin (70%), transthyretin (100%), RBP (85%), vitamins A (80%), B6 (37%), E (37%) and B12 (5%), zinc (20%) and copper (25%). Sixty-two per cent had two or more trace element or vitamin deficiencies. There was a weak association between micronutrient status and disease status. Micronutrient concentrations did not correlate with chronological age, height-for-age or weight for-age. CRP was elevated in 53% but did not correlate with any of the micronutrient concentrations. Micronutrient deficiencies were more common and micronutrient concentrations lower in children over 24 months of age. PMID- 11926046 TI - The impact of neonatal mortality on subsequent survival in rural Ethiopia. AB - In countries where most deliveries occur at home and most available information is hospital-based, accurate information on neonatal mortality is difficult to obtain. This study was conducted in a rural community in Ethiopia that has been under monthly demographic surveillance since 1987. The analysis in this paper was based on data collected in the 1st decade (1987-96) and this database was used to calculate mortality incidence rates and analyse survival. The overall neonatal mortality rate was 27/1000 live births (95% CI 24.5-29.5). The rates in the early and late neonatal periods were 20 and 8/1000 live births, respectively (95% CIs 18.0-22.9 and 6.6-9.4). The mortality incidence rates show that, every day, three of every 1000 newborns die in their 1st week of life. Neonatal mortality accounted for 43% of infant mortality. If all neonates survived the 1st week of life, life expectancy would increase by 1 year. Increased risk of neonatal mortality was found to be associated with living in a rural lowland area, twin births and male gender. This paper also addresses the need for further identification of the complex environmental and behavioural risk factors for neonatal mortality and for instituting appropriate and affordable interventions to reduce neonatal mortality. PMID- 11926047 TI - Fetal exposure to aflatoxins in the United Arab Emirates. AB - This is a prospective study in which aflatoxin levels were measured in umbilical cord blood from 201 women delivered consecutively in Tawam and Al Ain hospitals in order to determine whether the fetuses had been significantly exposed to the toxin. Aflatoxin B1, M1 and M2 were measured using high performance liquid chromatography. Aflatoxins were detected in 110 (54.7%) samples, 27 of which were positive for B1, 106 for M1 and 31 for M2. There was a significant negative correlation (p < 0.001) between birthweight and levels of aflatoxin. The high rate of detection of aflatoxins confirms that a significant number of infants in the UAE are exposed to these toxins which reflects maternal ingestion of aflatoxin-containing food. The presence of aflatoxin resulted in lower birthweights. PMID- 11926048 TI - A revised clinical method for assessment of severity of acute bacterial meningitis. AB - The aim of this study was to simplify a previously described clinical method of assessing severity of meningitis. An 8.5-point, six-item model for the risk of an abnormal course (seizures during treatment) or adverse outcome (death or recovery with neurological sequelae) was developed using a set of six bedside features: age < or = 2 yrs, 2 points; duration of illness > 7 days, 1.5 points; seizures, 2.5 points; hypovolaemic shock, 1 point; coma, 0.5 point; and abnormal muscle tone, 1 point. A high-risk score (< or = 2.5 points) was associated with a relative risk (95% CI) of 7.4 (2.4, 22.7) of seizures during treatment, and 6.3 (2.6, 17.2) for an adverse outcome (death or major or minor sequelae). The revised model should be suitable for use where laboratory facilities are not readily available, as in many developing countries, or when contra-indications to lumbar puncture are an important consideration on admission, as in severely ill patients, as well as when there are not such limitations. PMID- 11926049 TI - Sickle cell morbidity profile in Omani children. AB - In order to define the morbidity profile of sickle cell disease in Omani children, we analysed data on 97 children (53 boys, 44 girls) aged < or = 12 years admitted under our care in a regional referral hospital between July 1999 and June 2000. Ninety of them had sickle cell anaemia (HbSS disease) and seven had sickle cell thalassaemia (beta zero). Their mean (SD) steady-state Hb was 7.9 (1.2), range 6-10 g/dl. They were admitted on 316 occasions during the 12-month period. The number of admissions per child ranged from one to 12 (mean 3.3). Vaso occlusive crises were the main reason for admission (83%), followed by severe anaemia (12%) and infections (4%). During the study period, 31% received blood transfusions. Weight faltering was very common, 68% falling below the 5th percentile of the National Center for Health Statistics reference curves compared with 28% of age- and sex-matched non-sicklers (p < 0.001). Other complications included hypersplenism (four), ischaemic necrosis of the femoral head (two), and one case each of acute chest syndrome, acute splenic sequestration, cholelithiasis and pathological fracture of a lumbar vertebra. Overall, 71% of the children had moderately severe or severe disease. This pattern seems to be attributable, at least in part, to meteorological and genetic factors. The severe morbidity profile reported in this study underscores the need to continue the search for optimal management modalities, including the often emotion-laden issue of prevention. PMID- 11926051 TI - Mumps-like syndrome owing to parvovirus B19: a brief report. AB - Major manifestations of acute parvovirus B19 infection are usually erythema infectiosum, transient aplastic crisis and polyarthropathy. We report a case suspected to have mumps-meningo-encephalitis who had IgM and IgG antibodies positive for parvovirus B19. PMID- 11926050 TI - Cystic fibrosis in a child from Syria. AB - We describe a Syrian child with typical features of severe cystic fibrosis (CF) phenotype and a positive sweat test. DNA analysis confirmed homozygosity for the delta F508 mutation on chromosome 7. This report stresses the need to draw attention to and consider CF in Arab populations. The frequency and distribution of delta F508 in the Middle East are reviewed. PMID- 11926052 TI - Acute infantile haemorrhagic oedema associated with hepatitis A. AB - Acute infantile haemorrhagic oedema (AIHO) is characterised by purpura, ecchymosis and inflammatory oedema of the face and extremities. It is seen in children aged 4-24 months. The atiology is not known. We report a case of AIHO diagnosed by skin biopsy demonstrating leukocyte-elastic vasculitis. Laboratory studies showed positive hepatitis A IgM and IgG antibodies. The liver function tests were normal, indicating subclinical hepatitis. Cryoglobulinaemia was detected, suggesting that the disease was related to hepatitis A. PMID- 11926053 TI - Giardiasis as the cause of oedema and hypoproteinaemia in a child. AB - This report describes a 3-year-old child who presented with generalised oedema and hypoproteinaemia owing to giardiasis, was treated with oral metronidazole and recovered fully 10 days after therapy. The importance of considering giardiasis in patients with hypoproteinaemia of obscure aetiology is emphasised. PMID- 11926054 TI - Environmental risk factors in congenital malformations of the eye. AB - Developmental eye defects such as coloboma are a significant cause of visual morbidity in children, and are more common in India than elsewhere. The possible role of environmental factors in the aetiology of these conditions was investigated by studying birth order, symptoms of vitamin A deficiency (night blindness), drug use and maternal illness in pregnancy, rubella antibodies and exposure to agricultural chemicals. Through hospital records and community-based rehabilitation programmes in Andhra Pradesh, children with colobomata were recruited from schools for the blind. Eighty-three mothers of affected children were interviewed. The results showed that 43% of parents were consanguineous, that 19% had a positive family history and that the frequency of coloboma was highest in second-born children. Eleven (16%) mothers had a history of night blindness while pregnant with the affected child; seven (8%) took medication during the 1st trimester, abortifacients in two cases; three reported fever in the 1st trimester; and 11 (13%) reported exposure to agricultural chemicals. PMID- 11926055 TI - Lead levels in primary teeth of children in Karachi. AB - The majority of children in Karachi have blood lead levels above 10 micrograms/dl, the upper safety limit set by the United States Centers for Disease Control and Prevention. A study was undertaken to evaluate chronic lead exposure in children by measuring lead levels in shed primary teeth collected from primary school children in Karachi. A total of 309 teeth were collected from nine different schools, and analysed for lead content by atomic absorption spectrophotometry with electrothermal atomization. The mean (SD) age of the subjects was 7.6 (1.2) years and the mean lead level was 5.78 micrograms/g of whole tooth (dry weight), ranging from 0.42 to 39.75 micrograms/g. Incisor teeth had a significantly higher mean (SD) lead level, 6.42 (4.19) micrograms/g, than canines and molars which contained 4.91 (5.12) micrograms and 4.50 (2.67) micrograms lead whole tooth (dry weight), respectively. Significant differences were observed between different schools. No difference was observed between boys and girls and the accumulation of lead in teeth was not correlated with chronological age. PMID- 11926057 TI - Prolonged childhood Cushing's syndrome secondary to intralesional triamcinolone acetonide. AB - Intralesional corticosteroids have been used in the treatment of hypertrophic keloid scars for over 40 years. The few complications documented have mostly been local skin changes, viz depigmentation and cutaneous atrophy. Cushing's syndrome following intralesional injection is rare and usually self-limiting following cessation of therapy. This report describes a 9-year-old girl who developed Cushing's syndrome 2-3 weeks after a single treatment with 40 mg triamcinolone acetonide injected into keloids. Features of Cushing's syndrome and a suppressed hypothalamic-pituitary axis were present up to 9 months later. Three years after glucocorticoid injections, features of Cushing's syndrome had largely disappeared. Dosages of intralesional steroids normally recommended for adults are inappropriate for children. PMID- 11926056 TI - Effect of lactation counselling on subclinical mastitis among Bangladeshi women. AB - Subclinical mastitis, defined as raised milk sodium/potassium (Na/K) in the absence of clinical symptoms, is associated with poor infant weight gain and increased risk of mother-to-child HIV transmission. Similarly to clinical mastitis, subclinical mastitis appears to have multiple causes, one being poor lactation practice. We investigated whether a single session of lactation counselling around the time of delivery could decrease subclinical mastitis among women in rural Bangladesh. The counselling messages included the importance of giving colostrum, exclusive breastfeeding, feeding on demand and how to achieve good positioning and attachment. Women who had delivered prior to our instituting the counselling intervention (mean 1 month) constituted the uncounselled group. At 1 month postpartum, sodium and potassium were measured in spot milk samples from each breast by flame photometry and interleukin-8 (IL8) by ELISA. Geometric mean Na/K ratios and the proportion of milk samples with raised Na/K ratio were significantly (p < 0.05) lower in the counselled group [mean 0.42, 95% confidence interval (CI) 0.39-0.45, n = 116 samples; 10% Na/K ratio > 0.6] than in the non counselled group [mean 0.49, 95% CI 0.45-0.53, n = 127; 25% Na/K ratio > 0.6]. Milk IL8 concentration did not differ between groups. The results show that among Bangladeshi women a simple lactation counselling intervention could improve lactation practice in ways likely to improve infant health. PMID- 11926058 TI - Pattern of inborn errors of metabolism in an Omani population of the Arabian Peninsula. AB - We analysed all patients < 18 years of age diagnosed with inborn errors of metabolism (IEM) in the Metabolic Division, Sultan Qaboos University Hospital, Muscat, Oman from June 1998 to December 2000. A total of 82 patients from 76 families were studied, of whom 33 (40%) were aged < 1 week at presentation. Disorders identified included different lysosomal storage disorders (22), organic acidurias (9), carbohydrate metabolic disorders (9), congenital lactic acidosis (9), urea cycle disorders (8), amino acidopathies (8), fatty acid oxidation defects (7/82) and various other miscellaneous disorders (10). Tandem mass spectrometry helped in the diagnosis of 26 (32%) cases. Parental consanguinity was twice as frequent in the study patients as in the general population. Duration of follow-up ranged from 1 to 30 months (median 15) during which time 46/56 (82%) patients with disorders amenable to specific dietary and drug therapy available in Oman were free from frequent exacerbation. Our study shows the relevance of identifying patients with IEM in Oman and the need to establish screening for the conditions identified and provide effective management protocols. PMID- 11926059 TI - Appendicitis-like syndrome owing to mesenteric adenitis caused by Salmonella typhi. AB - We report a 14-year-old girl who presented with signs of appendicitis and had her appendix removed. She subsequently proved to have mesenteric adenitis owing to Salmonella typhi which responded to treatment with ceftriaxone. PMID- 11926060 TI - LIM proteins: association with the actin cytoskeleton. AB - The LIM domain is an evolutionary conserved double-zinc finger motif found in a variety of proteins exhibiting diverse biological roles. LIM domains have been observed to act as modular protein-binding interfaces mediating protein-protein interactions in the cytoplasm and the nucleus. Interaction of LIM domains with specific protein partners is now known to influence its subcellular localization and activity; however, no single binding motif has been identified as a common target for LIM domains. Several LIM domain-containing proteins associated with the actin cytoskeleton have been identified, playing a role in signal transduction and organization of the actin filaments during various cellular processes. PMID- 11926061 TI - Xylan deposition on secondary wall of Fagus crenata fiber. AB - Delignified and/or xylanase-treated secondary walls of Fagus crenata fibers were examined by field emission scanning electron microscopy. Microfibrils with a smooth surface were visible in the innermost surface of the differentiating fiber secondary wall. There was no ultrastructural difference between control and delignified sections, indicating that lignin deposition had not started in the innermost surface of the cell wall. There was no ultrastructural difference between control and xylanase-treated sections. Microfibrils on the outer part of the differentiating secondary wall surface had globular substances in delignified sections. These globular substances disappeared following xylanase treatment, indicating that these globules are xylan. The globular substances were not visible near the inner part of the differentiating secondary wall but gradually increased toward the outer part of the secondary wall, indicating that xylan penetrated into the cell wall and continuously accumulated on the microfibrils. Mature-fiber secondary walls were also examined by field emission scanning electron microscopy. Microfibrils were not apparent in the secondary wall in control specimens. Microfibrils with many globular substances were observed in the delignified specimens. Following xylanase treatment, the microfibrils had a smooth surface without any globules, indicating that the globular substance is xylan. These results suggest that cellulose microfibrils synthesized on the plasma membrane are released into the innermost surface of the secondary wall and coated with a thin layer of xylan. Successive deposition of xylan onto the cell wall increases the microfibril diameter. The large amounts of xylan that accumulated on microfibrils appear globular but are covered with lignin after they are deposited. PMID- 11926062 TI - Spatio-temporal relationship between nuclear-envelope breakdown and preprophase band disappearance in cultured tobacco cells. AB - Cell division involves the coordinated progression of karyokinesis and cytokinesis, which is accomplished by communication between the nucleus and the cytoplasm. We have utilized green-fluorescent-protein technology to generate a line of tobacco 'Bright Yellow 2' (BY-2) cells labeled for both microtubules and the nuclear envelope. This cell line allowed us to use living cells to investigate the relationship between nuclear-envelope breakdown and preprophase band disappearance with high spatial and temporal resolution. Our observations demonstrate that nuclear-envelope breakdown always precedes preprophase band disappearance in BY-2 cells. In addition, the rate of preprophase band disappearance, and the attenuation of perinuclear microtubule fluorescence, correlates with the proximity of the nucleus to the preprophase band site. These results indicate the presence of communication between the nucleus and the preprophase band and suggest a causal relationship between nuclear-envelope breakdown and preprophase band disappearance. PMID- 11926063 TI - Cytoskeletal responses during early development of the downy mildew of grapevine (Plasmopara viticola). AB - A host-free system was established to induce the early development of the obligate biotrophic pathogen Plasmopara viticola, the downy mildew of grapevine. This system was used to study cytoskeletal responses during encystation and germ tube formation. During these processes, both the actin and the tubulin cytoskeleton show a stage-specific pattern of distribution. Elimination of the cytoskeleton by the actin drug latrunculin B and the microtubule drug ethyl-N phenyl-carbamate did not affect the release of mobile zoospores from the sporangia, nor the encystation process, but efficiently inhibited the formation of a germ tube. The data are discussed with respect to a role of both actin and microtubules for the establishment of the cell polarity guiding the emergence and the growth of the germ tube. PMID- 11926064 TI - Immunocytochemical evidence of calreticulin-like protein in pollen tubes and styles of Petunia hybrida Hort. AB - With a polyclonal antibody raised against calreticulin (CRT) the locations where the protein occurs in unpollinated and pollinated styles of Petunia hybrida were localized. The epitopes binding the CRT antibody were immunolocalized preferentially in pollen tubes. In transmitting tract cells, both before and after pollination, the level of CRT was low. The protein was mainly localized in the cytosol and around dictyosomes of transmitting-tract cells. In pollen tubes, a high level of CRT was found at their tips rich in endoplasmatic reticulum, cisternae piles of reticular and/or dictyosomal origin, and vesicles. Binding sites of the CRT antibody were also found in the internal callosic cell wall of the pollen tube. These results indicate a role of CRT in cells directly participating in pollen-pistil interaction. PMID- 11926065 TI - The pore of the leaf cavity of Azolla species: teat cell differentiation and cell wall projections. AB - The differentiation of the specialized secretory teat cells of the leaf cavity pore of Azolla species was investigated at the ultrastructural level with emphasis on their peculiar cell wall projections. The results indicated that the projections are formed as soon as the teat cells complete their differentiation and that their production is principally associated with changes in endoplasmic reticulum profiles. The number of projections increases with the teat cell age and is stimulated under salt and P deficiency stresses. Salt stress also promotes their emergence on Azolla species that under normal conditions do not produce projections. Cytochemical tests on different Azolla species showed that the projection composition is almost identical: proteins, acidic polysaccharides, and pectin are always detected. This study revealed that Azolla teat cell projections differ fundamentally from other types of hitherto described cell wall projections that are considered as remnant structures from cell separation. In contrast, in Azolla teat cells projections are actively produced and compounds are excreted by an exocytotic mechanism. The possible role of the projections in the symbiosis of Azolla spp. with Anabaena azollae is discussed. PMID- 11926066 TI - Epidermal cells of a symbiosis-defective mutant of Lotus japonicus show altered cytoskeleton organisation in the presence of a mycorrhizal fungus. AB - The influence of the mycorrhizal fungus Gigaspora margarita on cytoskeleton organisation in epidermal cells of Lotus japonicus roots was compared between plants of the wild type Gifu and the mutant Ljsym4-2, in which the fungus is confined to the epidermal cells. Immunofluorescence labelling of plant microtubules and microfilaments showed only limited alterations in the peripheral cytoskeleton of epidermal cells during early stages of fungal interaction with the wild type. Later, microtubules and microfilaments enveloped the growing hypha, while the host cell nucleus moved close to the fungus. In contrast, epidermal cells of the mutant responded with disorganisation and disassembly of microtubules and microfilaments before and during fungal penetration attempts. The fungus penetrated only as far as to epidermal cells, whose cytoplasm became devoid of tubulin and actin, suggesting cell death. The close relationship between host cytoskeleton organisation and compatibility with the fungus suggests that a functional Ljsym4 gene is necessary for correct reorganisation of the epidermal cell cytoskeleton in the presence of the fungus and for avoiding hypersensitivity-like reactions. PMID- 11926067 TI - Identification of the YMN-1 antigen protein and biochemical analyses of protein components in the mitochondrial nucleoid fraction of the yeast Saccharomyces cerevisiae. AB - We analyzed the protein components contained in the mitochondrial nucleoid (mt nucleoid) fraction of the yeast Saccharomyces cerevisiae. Immunoblotting with anti-Abf2p antibody demonstrated the association of Abf2p, a major mitochondrial DNA-binding protein, with the mt-nucleoids. In contrast, porin and cytochrome c oxidase subunit III (CoxIIIp) were not detected by immunoblotting in the mt nucleoid fraction. The YMN-1 monoclonal antibody recognized a 48 kDa protein of the mt-nucleoid fraction. The N-terminal amino acid sequence of the protein and immunological evidence showed that the YMN-1 monoclonal antibody recognizes dihydrolipoyl transsuccinylase (KE2), which is one of the constituents of the alpha-ketoglutarate dehydrogenase complex (KGDC). alpha-Ketoglutarate dehydrogenase (KE1) and dihydrolipoyl dehydrogenase (E3), which are other subunits of KGDC, were also detected in the mt-nucleoid fraction. An enzyme assay of the mt-nucleoid fraction showed that cytochrome c oxidase and fumarase activity were barely detected in the fraction, but the specific activity of KGDC in the mt-nucleoid fraction was relatively high and was approximately 60% of the specific activity in the mitochondrial fraction. Three components of KGDC were detected in the DNA-binding protein fractions after DNA-cellulose column chromatography of mt-nucleoid proteins. These results suggested that a part of KGDC in the mitochondrial matrix is associated with mt-nucleoids in vivo. PMID- 11926069 TI - A role for actin-driven secretion in auxin-induced growth. AB - In epidermal cells of Zea mays coleoptiles, actin microfilaments are organized in fine strands during cell elongation, but are bundled in response to signals that inhibit growth. This bundling response is accompanied by an increased membrane association of extracted actin. Brefeldin A, an inhibitor of vesicle secretion, increases the membrane association of actin, causes a bundling of cortical actin microfilaments, and reduces the sensitivity of cell elongation to auxin. A model is proposed where auxin controls the dynamics of an actin subpopulation that guides vesicles loaded with components of the auxin-signaling machinery towards the cell poles. PMID- 11926068 TI - A comparative ultrastructural study of pollen development in Arabidopsis thaliana ecotype Columbia and male-sterile mutant apt1-3. AB - Adenine phosphoribosyltransferase (APT) catalyzes the conversion of adenine and cytokinin bases to the corresponding nucleotides. An Arabidopsis thaliana mutant lacking the major APT isoform, APT1, is male sterile due to defects soon after meiosis. We have now used electron microscopy to define the effects of APT1 deficiency on pollen development to determine whether the changes might be attributed to adenine or cytokinin metabolism. Changes were observed in mutant anthers in both tapetal and pollen mother cells prior to meiosis with additional defects found at later stages, in both compartments. Principal changes include altered lipid accumulation in the tapetal cells, changes in pollen cell wall development, and a loss of synchrony in the development of the tapetum and microspores. Taken together our results suggest that APT1 deficiency causes a general metabolic decrease in energy metabolism, due to the lack of adenine recycling into adenylate nucleotides, which ultimately leads to pollen abortion. The early onset of meiosis in the mutant may be associated with altered cytokinin metabolism. PMID- 11926070 TI - Sesquiterpene lactone dehydroleucodine selectively induces transient arrest in G2 in Allium cepa root meristematic cells. AB - Dehydroleucodine is a sesquiterpene lactone recently isolated from aerial parts of a medicinal herb, Artemisia douglasiana Besser. We have previously shown that 25 and 100 microM dehydroleucodine slowed down onion root growth by 30 and 70%, respectively, affecting neither cell viability nor cell elongation. In the present study we analyze the effect of dehydroleucodine on cell cycle phases in onion (Allium cepa L.) root meristematic cells synchronized with caffeine or caffeine and hydroxyurea. Synchronized root cells treated with 100 and 200 microM dehydroleucodine present an interphase lengthening of 5.2 h and 8.2 h, respectively. The S-phase length, estimated by [3H]thymidine incorporation assay, was 6 h for both control roots and roots that had been immersed in dehydroleucodine. The peak of [3H]leucine incorporation was observed 6 h after release from synchronization in controls and in dehydroleucodine-treated roots, indicating that protein synthesis in G2 was not affected. Thus, these results show that dose-dependently dehydroleucodine selectively induces a transient arrest of meristematic cell in G2 and that dehydroleucodine can be used experimentally as a cell cycle suppressor. PMID- 11926071 TI - Arabinogalactan proteins, pollen tube growth, and the reversible effects of Yariv phenylglycoside. AB - Arabinogalactan proteins (AGPs) are abundant complex macromolecules involved in both reproductive and vegetative plant growth. They are secreted at pollen tube tips in Lilium longiflorum. Here, we report the effect of the (beta-D-glucosyl)3 Yariv phenylglycoside, known to interact with AGPs, on pollen tube extension in several plant species. In Annona cherimola the Yariv reagent clearly inhibited pollen tube extension within 1-2 h of treatment, as demonstrated previously for L. longiflorum, but had no effect on Lycopersicon pimpinellifolium, Aquilegia eximia, and Nicotiana tabacum. With the monoclonal antibody JIM13 we also examined these same species for evidence that they secreted AGPs at their pollen tube tips. Only A. cherimola showed evidence of AGPs at the pollen tube tip as does lily. The Yariv reagent causes arrest of tube growth in both A. cherimola and lily, but its removal from the medium allows regeneration of new tip growth in both species. We show that the site of the new emerging tip in lily can be predicted by localization of AGP secretion. Labeling with JIM13 appeared on the flanks of the arrested tip 1 h after removal of the Yariv reagent from the growth medium. After 4 h, many of the Yariv reagent-treated pollen tubes had regenerated new pollen tubes with the tips brightly labeled by JIM13 and with a collar of AGPs left at the emergence site. During this recovery, esterified pectins colocalized with AGPs. Secretion at the site of the new tip may be important in the initial polarization event that occurs on the flanks of the arrested tube tip and results in a new pollen tube. PMID- 11926072 TI - Observation of polarity induction by cytochemical localization of phenylalkylamine-binding sites in regenerating protoplasts of the moss Physcomitrella patens. AB - Different external (e.g., light) and internal (e.g., auxin and calcium gradients) factors control differentiation of the moss protonema. The present investigations demonstrate that exogenously applied auxin, the pharmacological blockade of auxin efflux by naphthylphthalamic acid, and treatment with (-)bepridil, a calcium channel antagonist, inhibit protoplast division without affecting protoplast viability in the moss Physcomitrella patens. A fluorescently labelled phenylalkylamine (DM-Bodipy PAA), another calcium channel antagonist, was used as a probe for in vivo labelling of phenylalkylamine(PAA)-binding sites. The specificity of this binding was demonstrated by competition with (-)bepridil. Confocal laser scanning microscopy visualized PAA-binding sites on the plasma membrane and along the nuclear membrane as uniformly distributed clusters. During asymmetric division of P. patens protoplasts, however, fluorescence labelling particularly increases at the membrane invagination and later along the plate separating the new cells. Intracellular localization of PAA-binding sites, probably at the membranes of vesicles and vacuoles, significantly increases in the smaller daughter cell, destined to later form a polar outgrowth, the first chloronema cell. Thus, a system was established to visualize early events in P. patens protoplast polarization at the subcellular level. PMID- 11926073 TI - Mood disorders--new definitions, treatment directions, and understanding. PMID- 11926074 TI - "Cade's disease" and beyond: misdiagnosis, antidepressant use, and a proposed definition for bipolar spectrum disorder. AB - The diagnosis and treatment of bipolar disorder (BD) has been inconsistent and frequently misunderstood in recent years. To identify the causes of this problem and suggest possible solutions, we undertook a critical review of studies concerning the nosology of BD and the effects of antidepressant agents. Both the underdiagnosis of BD and its frequent misdiagnosis as unipolar major depressive disorder (MDD) appear to be problems in patients with BD. Underdiagnosis results from clinicians' inadequate understanding of manic symptoms, from patients' impaired insight into mania, and especially from failure to involve family members or third parties in the diagnostic process. Some, but by no means all, of the underdiagnosis problem may also result from lack of agreement about the breadth of the bipolar spectrum, beyond classic type I manic-depressive illness (what Ketter has termed "Cade's Disease"). To alleviate confusion about the less classic varieties of bipolar illness, we propose a heuristic definition, "bipolar spectrum disorder." This diagnosis would give greater weight to family history and antidepressant-induced manic symptoms and would apply to non-type I or II bipolar illness, in which depressive symptom, course, and treatment response characteristics are more typical of bipolar than unipolar illness. The role of antidepressants is also controversial. Our review of the evidence leads us to conclude that there should be less emphasis on using antidepressants to treat persons with this illness. PMID- 11926076 TI - Major depression and its association with long-term medical conditions. AB - OBJECTIVE: To replicate previously reported associations between major depressive episodes (MDEs) and long-term medical conditions in a Canadian community sample. METHODS: A sample of 2542 household residents was selected using random digit dialing (RDD). Data were collected by telephone interview. The Composite International Diagnostic Interview (CIDI)-Short Form for major depression (CIDI SFMD) was used to identify MDEs occurring in the previous 12 months. Long-term medical conditions were identified by self-report. RESULTS: The prevalence of MDE was elevated in those subjects who reported 1 or more long-term medical conditions. The association was not due to confounding by age, sex, social support, or stressful recent life events. CONCLUSION: This study replicates a previously reported association between depressive disorders and long-term medical conditions. These cross-sectional associations suggest that medical conditions may increase the risk of major depression or that major depression may increase the risk of medical conditions. Alternatively, comorbid medical conditions may influence the duration of depressive episodes, or vice versa. These explanations are not mutually exclusive. PMID- 11926075 TI - The neurobiology of bipolar disorder: focus on signal transduction pathways and the regulation of gene expression. AB - OBJECTIVE: This article presents an overview of signal transduction pathways and reviews the research undertaken to study these systems in clinically relevant samples from patients with bipolar disorder (BD). METHOD: We reviewed the published findings from studies of postmortem brain tissue and blood samples from patients with BD. RESULTS: Although the exact biochemical abnormalities have yet to be identified, the presented findings strongly suggest that BD may be due, at least in part, to abnormalities in signal transduction mechanisms. In particular, altered levels or function, or both, of G-protein alpha subunits and effector molecules such as protein kinase A (PKA) and protein kinase C (PKC) have consistently been associated with BD both in peripheral cells and in postmortem brain tissue, while more recent studies implicate disruption in novel second messenger cascades, such as the ERK/MAPK pathway. CONCLUSIONS: Despite the difficulties inherent in biochemical studies of clinically relevant tissue samples, numerous investigations have illuminated the signal transduction mechanisms in patients with BD. These studies also suggest that BD may be due to the interaction of many abnormalities. In this context, novel techniques enabling the study of gene expression promise to assist in untangling these complex interactions, through visualizing the end result of these changes at the level of gene transcription. PMID- 11926077 TI - Seasonal affective disorders: relevance of Icelandic and Icelandic-Canadian evidence to etiologic hypotheses. AB - OBJECTIVE: This study tests the suggestion of earlier studies concerning the importance of genetic factors in the etiology of winter seasonal affective disorders (SADs) and subsyndromal winter SAD (S-SAD). METHOD: Two study populations of Winnipeg, Manitoba residents were canvassed: 250 adults of wholly Icelandic descent and 1000 adults of non-Icelandic descent. We distributed the Seasonal Pattern Assessment Questionnaire by mail to these 2 populations, yielding 204 and 449 valid responses, respectively. RESULTS: Rates of SAD and S SAD proved markedly lower in the Icelandic population than those in the non Icelandic population. CONCLUSIONS: These differences seem unexplained by differences in ambient light or climate, thus indicating that genetic factors contribute to the expression of SADs. Compared with earlier findings from a group of adults of wholly Icelandic descent living in nearby rural Manitoba, the etiologic importance of as-yet-undetermined environmental factors unrelated to latitude or ambient light is also indicated. PMID- 11926078 TI - Canadian psychiatric inpatient religious commitment: an association with mental health. AB - BACKGROUND: Research indicates that religion may have a positive effect on coping and possibly enhance clinical outcomes. This study aims to determine the level of religious interest of psychiatric inpatients and to assess whether religious commitment has an impact on selected outcome variables. METHODS: There were 88 consecutive adult patients (50% men) who were admitted to a Canadian tertiary care psychiatry inpatient unit and were interviewed about their religious beliefs and practices. Patients with a Beck Depression score of 12 or more were included for outcome analysis. RESULTS: A total of 59% believed in a God who rewards and punishes, 27% had a high frequency of worship attendance, and 35% prayed once or more daily. More frequent worship attenders had less severe depressive symptoms, shorter current length of stay, higher satisfaction with life, and lower rates of current and lifetime alcohol abuse (P < 0.05), when compared with those with less frequent or no worship attendance. In contrast, private spirituality was associated with lower depressive symptoms and current alcohol use only (P < 0.05), and prayer frequency had no significant associations. DISCUSSION: This study indicates that certain religious practices may protect against severity of symptoms, hospital use, and enhance life satisfaction among psychiatric inpatients. This is the first known Canadian study that examines religious commitment among psychiatric inpatients. PMID- 11926079 TI - The moderating effects of coping strategies on major depression in the general population. AB - OBJECTIVES: To evaluate the moderating effects of various coping strategies on the association between stressors and the prevalence of major depression in the general population. METHODS: Subjects from the Alberta buy-in component of the 1994-1995 National Population Health Survey (NPHS) were included in the analysis (n = 1039). Each subject was asked 8 questions about coping strategies that dealt with unexpected stress from family problems and personal crises. Major depression was measured using the World Health Organization's (WHO) Composite International Diagnostic Interview-Short Form (CIDI-SF) for major depression. The impacts of coping strategies in relation to psychological stressors on the prevalence of major depression were determined by examining interactions between coping and life stress on major depression using logistic regression modelling. RESULTS: No robust impact of coping strategies in relation to various categories of stress evaluated in the NPHS was observed. There was evidence that the use of "pray and seek religious help" and "talks to others about the situations" as coping strategies by women moderated the risk of major depression in the presence of financial stress and relationship stress (with a partner). Using emotional expression as a coping strategy by women might decrease the risk of major depression in the presence of 1 or more recent life events, personal stress, relationship stress (with a partner), and environmental stress. CONCLUSION: Different coping strategies may have a differential impact on the prevalence of major depression in specific circumstances. These findings may be important both to prevent and to treat depressive disorders. PMID- 11926080 TI - Antidepressant side effects in depression patients treated in a naturalistic setting: a study of bupropion, moclobemide, paroxetine, sertraline, and venlafaxine. AB - OBJECTIVE: There is no commonly accepted standard for comparing antidepressant induced side effects. This study evaluates a clinician-administered scale, the Toronto Side Effect Scale (TSES), in a natural practice clinic. METHOD: We used the TSES to assess side effects in 193 depression patients who completed 8 weeks of treatment with either bupropion, moclobemide, paroxetine, sertraline, or venlafaxine. RESULTS: Rates of remission (Hamilton Rating Scale for Depression [HRSD] < 7) did not differ across drugs after 8 weeks of treatment. Paired drug comparisons yielded significant differences in 16 of the 32 side effects. We present differences between pairs of the 5 antidepressants in Central Nervous System (CNS), gastrointestinal (GI), and sexual side effects. A measure of side effect intensity distinguished paroxetine from the other antidepressants on a measure of sexual dysfunction. CONCLUSIONS: These results confirm the clinical utility of the TSES as a simple, clinician-administered antidepressant side effect scale. PMID- 11926082 TI - Short-term dynamic psychotherapies in the treatment of major depression. PMID- 11926081 TI - Treatment delays for involuntary psychiatric patients associated with reviews of treatment capacity. AB - OBJECTIVE: To determine the duration of delays in treatment initiation when involuntary patients apply for a review of a finding of treatment incapacity and to estimate the cost of keeping patients hospitalized without treatment in these circumstances. METHOD: Using a computerized database and written records, we identified all patients at 2 psychiatric hospitals in Ontario who applied for a review of a finding of treatment incapacity during a 10-year period. We recorded clinical and demographic variables, dates of stopping and starting medication, and dates of review board hearings and outcomes. We also noted all cases in which a patient appealed a decision from the review board to the court. RESULTS: Two hundred and thirty-seven patients made 334 applications to the review board. The board overturned the physician's finding of incapacity in only 5 (1.5%) applications; 15 appealed the review board's finding to the courts. None of these appeals were successful. In the absence of an appeal to the courts, the average delay in initiating treatment was 25 days. For patients appealing to the court, the average delay was 253 days. The cost of hospitalizing untreated patients while their capacity was under legal review was estimated at $3,867,000, of which $1,333,000 could have been saved if treatment had started immediately after the review board confirmed incapacity. CONCLUSION: We have identified extensive delays in initiating psychiatric treatment for a number of patients. These delays are associated with legal review of treatment capacity. There are serious clinical risks and substantial costs associated with delay in treating patients with acute psychiatric illness. Where jurisdictions review treatment capacity, we recommend that treatment not be impeded once a review board has confirmed a clinical finding of incapacity. PMID- 11926083 TI - Using financial incentives to promote shared mental health care. PMID- 11926084 TI - Borderline personality disorder comorbidity in early- and late-onset bipolar II disorder. PMID- 11926085 TI - DAD = DSM-IV 296.23. PMID- 11926086 TI - Relation of personality disorders to subtypes of major depression according both to DSM-IV and ICD-10. PMID- 11926087 TI - Worry WARTS have generalized anxiety disorder. PMID- 11926088 TI - Getting returns for sender. PMID- 11926089 TI - CIOs double as change agents. PMID- 11926090 TI - Getting along in the 'C-suite'. PMID- 11926091 TI - Defining the roles of HIPAA officers. PMID- 11926092 TI - I.T. talent: in or out? PMID- 11926093 TI - Readers perspectives. Though hand-held computers increasingly are being used in clinical settings, the technology's limitations will limit the use.. PMID- 11926094 TI - Talking with Mark Smits of Abbott Diagnostics. Interview by C. Anne Pontius. PMID- 11926095 TI - C-reactive protein: from acute phase reactant to cardiovascular disease risk factor. PMID- 11926096 TI - Integrated networks look at pros and cons of dropping their GPOs. PMID- 11926097 TI - OR design & construction. Healing environments in surgical suites. PMID- 11926098 TI - OR design & construction. What do we know about healing environments? PMID- 11926099 TI - MDs, CRNAs debate value of assistants. PMID- 11926101 TI - Planning for successful successions. PMID- 11926100 TI - Don't use compressed air to clean scopes. PMID- 11926102 TI - Emergency patients come to the ASC. PMID- 11926103 TI - 3-month course prepares outpatient RNs. PMID- 11926104 TI - Making pain management a viable service. PMID- 11926105 TI - VHA's program to curb wrong-site surgery. PMID- 11926106 TI - Case managers must take a proactive role in managing denials. PMID- 11926107 TI - Software can improve denial management. PMID- 11926108 TI - Effective documentation: know you're doing it right. PMID- 11926109 TI - Medication errors: how CMs can make care safer. Part 1. PMID- 11926110 TI - Critical path network. New protocol can rule out MIs in 90 minutes. PMID- 11926111 TI - Discharge process enables faster placement. PMID- 11926112 TI - To smooth discharge, check bed 'life cycle'. PMID- 11926113 TI - Patient safety alert. Early results of leapfrog hospital survey promising. PMID- 11926114 TI - Patient safety alert. Safety tool stresses education and action. PMID- 11926115 TI - Diffusion approximation of the stochastic process of microtubule assembly. AB - Microtubules are protein polymers that guide intracellular motility. Stochastic switching of a microtubule between states of elongation, shortening, and pause is described in detail by the dynamic instability (DI) model. Recently we have described the dynamics of microtubules phenomenologically as generalized diffusion of their ends. Genesis of the diffusion dynamics and accuracy of diffusion model are studied in this work. It is shown that wandering of the end of a microtubule undergoing DI asymptotically approaches the Wiener diffusion process. Accuracy of the diffusion approximation is evaluated by comparing its predictions with results of simulation of DI. Stationary distributions of microtubule length and life-time that are predicted by both models differ qualitatively between two cell types considered. However, predictions of the diffusion model are in each case practically identical to predictions of the DI model being also consistent with experimental data. The peculiar stochastic process of microtubule assembly thus converges at cell scale to a kind of widespread-in-nature diffusion process. This result is considered an example of qualitative change in dynamical properties in transition from the molecular to cellular level of biological organization. Additionally, it suggests employment of diffusion process theory in studying functions of microtubules in the cell. PMID- 11926116 TI - A mathematical model of partial-thickness burn-wound infection by Pseudomonas aeruginosa: quorum sensing and the build-up to invasion. AB - Pseudomonas aeruginosa remains a significant pathogen in burn-wound infection, its pathogenicity being associated with the production of a cocktail of virulence determinants which is regulated by a population-density-dependent mechanism termed quorum sensing. Quorum sensing is effected through the production and binding of signalling molecules. Here we present a mathematical model for the early stages of the infection process by P. aeruginosa in burn wounds which accounts for the quorum sensing system and for the diffusion of signalling molecules in the burn-wound environment. The results of the model and the effects of important parameters are discussed in detail. For example, the effect of the degradation rate of signalling molecules and its significance for anti-signalling therapies is discussed. PMID- 11926117 TI - Home range formation in wolves due to scent marking. AB - Social carnivores, such as wolves and coyotes, have distinct and well-defined home ranges. During the formation of these home ranges scent marks provide important cues regarding the use of space by familiar and foreign packs. Previous models for territorial pattern formation have required a den site as the organizational center around which the territory is formed. However, well-defined wolf home ranges have been known to form in the absence of a den site, and even in the absence of surrounding packs. To date, the quantitative models have failed to describe a mechanism for such a process. In this paper we propose a mechanism. It involves interaction between scent marking and movement behavior in response to familiar scent marks. We show that the model yields distinct home ranges by this new means, and that the spatial profile of these home ranges is different from those arising from the territorial interactions with den sites. PMID- 11926118 TI - A dual-mode dynamic model of the human accommodation system. AB - The function of the accommodation system is to provide a clear retinal image of objects in the visual scene. The system was previously thought to be under simple continuous (i.e., single mode of operation) feedback control, but recent research has shown that it is under discontinuous (i.e., two stimulus-dependent modes of operation) feedback control by means of fast and slow processes. A model using MATLAB/SIMULINK was developed to simulate this dual-mode behavior. It consists of fast and slow components in a feedback loop. The fast component responds to step target disparity with an open-loop movement to nearly reach the desired level, and then the slow component uses closed-loop feedback to reduce the residual error to an acceptable small level. For slow ramps, the slow component provides smooth tracking of the stimulus, whereas for fast ramps, the fast component provides accurate staircase-like step responses. Simulation of this model using a variety of stimuli, including pulse, step, ramp, and sinusoid, showed good agreement with experimental results. Thus, this represents the first dynamic model of accommodation that can accurately simulate the complex dual-mode behavior seen experimentally. The biological significance of this model is that it can be used to quantitatively analyze clinical deficits such as amblyopia and accommodative insufficiency. PMID- 11926119 TI - Modelling, steady state analysis and optimization of the catalytic efficiency of the triosephosphate isomerase. AB - In the present work we have modelled and optimized the reaction mechanism of the triose phosphate isomerase (TIM) enzyme (E.C. 5.3.1.1). For this purpose we have used an approach that combines the S-system representation within the power law formalism and linear programming techniques. By this means we have explored those rate constants whose alterations are likely to improve the catalytic efficiency of the enzyme and investigated the available room for optimization in different metabolic conditions. The role and plausibility of the different types of mutations on the evolution of this enzyme have also been considered. Steady state sensitivity analysis was carried out and a new set of aggregated logarithmic gains was defined in order to quantify the responses of the system to changes in groups of rate constants that could be explained in terms of mutations affecting the catalytic properties of the enzyme. Evaluation of these logarithmic gains at different levels of saturation and disequilibrium ratios enabled us to reach conclusions about the meaning and role of the diffusion limitation terms. The catalytic efficiency of the monoenzymatic system was optimized through changes in the kinetic rate constants within different sets of restrictions ranging from thermodynamic or kinetic to evolutionary ones. Results showed that, at very different conditions, there is still room for improvement in the TIM enzyme. Thus, in a wide range of metabolically significant values of the disequilibrium ratio there is a minimal variation in the optimal profile that yields 2.1 times the velocity of the basal states. Though most of this increase is accounted for by the increase of the second order constants (that could have already reached a theoretical maximum) significant increases (20%) in catalytic efficiencies are obtained by changes of the internal steps only. Besides these new findings our optimization approach has been able to reproduce results obtained with other approaches. PMID- 11926120 TI - Modelling Dictyostelium discoideum morphogenesis: the culmination. AB - The culmination of the morphogenesis of the cellular slime mould Dictyostelium discoideum involves complex cell movements which transform a mound of cells into a globule of spores on a slender stalk. We show that cyclic AMP signalling and differential adhesion, combined with cell differentiation and slime production, are sufficient to produce the morphogenetic cell movements which lead to culmination. We have simulated the process of culmination using a hybrid cellular automata/partial differential equation model. With our model we have been able to reproduce the main features that occur during culmination, namely the straight downward elongation of the stalk, its anchoring to the substratum and the formation of the long thin stalk topped by the spore head. We conclude that the cyclic AMP signalling system is responsible for the elongation and anchoring of the stalk, but in a roundabout way: pressure waves that are induced by the chemotaxis towards cyclic AMP squeeze the stalk through the cell mass. This mechanism forces the stalk to elongate precisely in the direction opposite to that of the chemotactically moving cells. The process turns out to be 'guided' by inactive 'pathfinder' cells, which form the tip of the stalk. We show that the entire development is enacted by means of the aforementioned building blocks. This means that no global gradients or different modes of chemotaxis are needed to complete the culmination. PMID- 11926121 TI - Kinetic heterogeneity of an experimental tumour revealed by BrdUrd incorporation and mathematical modelling. AB - In the present paper we propose a method of analysis of the cell kinetic characteristics of in vivo experimental tumours, that uses DNA-BrdUrd flow cytometry data at various times after the bromodeoxyuridine (BrdUrd) injection and mathematical modelling. The model of the cell population takes into account the cell-cell heterogeneity of the progression rate across cell cycle phases within the tumour, and assumes a strict correlation between the durations of S and G2M phases. The model also allows for a nonconstant DNA synthesis rate across S phase. In addition, the measurement process is modelled, considering the possibility of nonimpulsive labelling and providing a representation of the time course of the bivariate DNA-BrdUrd fluorescence distribution. Sequential DNA BrdUrd distributions were obtained in vivo from a human ovarian carcinoma transplanted in mice and, for comparison, in vitro from a cell line of the same origin. From these data, that included the fractional density and the mean BrdUrd fluorescence of BrdUrd-positive cells as a function of the DNA-fluorescence, kinetic parameters such as the potential doubling time (Tpot) and the mean and variance of the transit times in S and G2M phases, were estimated. This study revealed the presence of a substantial heterogeneity in S and G2M phases within the in vivo cell population and of a lower heterogeneity in the in vitro population. Moreover, our analysis suggests a nonnegligible effect of the BrdUrd pharmacokinetics in the in vivo cell labelling. PMID- 11926122 TI - Modeling deuterated glucose labeling of T-lymphocytes. AB - Human immunodeficiency virus type 1 (HIV-1) infects cells of the immune system and leads to depletion of CD4+ T cells, and to an increase of CD8+ T-lymphocytes. However, not much is known about the dynamics of turnover (proliferation and death) of the CD4+ and CD8+ T cell populations in HIV-infected and healthy individuals. A new experimental technique has been developed using deuterated glucose labeling that provides information on cell turnover in vivo. However, the quantitative interpretation of the data requires the development of specific dynamic models. In this paper we derive two models, a simple one-compartment model and a more complex two-compartment model. These models allow for robust quantification of death and proliferation rates, but careful consideration of the system is necessary to understand what is being measured in each case. We demonstrate that more realistic models can account not only for differences in the turnover rates between HIV-infected and healthy individuals, but also take into consideration the elevated state of activation in HIV infection. The use of these models in the interpretation of the experimental data will increase our knowledge of T cell dynamics in the context of HIV infection. PMID- 11926123 TI - Stochasticity of the step size in the force generating process due to a single myosin molecule. AB - We formulate a motional model on the basis of an experimental optical trapping system which reveals the single molecular events of the force generation due to actin and myosin, and we discuss the origin of the dispersion of the displacements of the beads bound to the actin filament. In some experimental data sets, this dispersion is larger than that presumed from the model under the condition that the thermal agitation is the only source of the fluctuation. Thus, other fluctuating elements besides that due to thermal agitation are supposed to cause the dispersion. A possible fluctuating element is the step size itself. We discuss this possibility and estimate the fluctuation in step size using two reported experimental data sets. PMID- 11926124 TI - Web alert. The chemistry of biological products. PMID- 11926126 TI - The genus Bifidobacterium for cancer gene therapy. AB - A fundamental obstacle in cancer gene therapy is the specific targeting of therapy to solid tumor; as yet, no systemic delivery system exists. Bifidobacterium longum, a strain of domestic bacteria that is non-pathogenic and anaerobic, selectively localized to and proliferated in solid tumors after systemic application. We propose a novel approach to cancer gene therapy in which anaerobic bacteria of the genus B longum are used to achieve tumor-specific gene delivery and enzyme-prodrug therapy. PMID- 11926125 TI - Live bacteria as anticancer agents and tumor-selective protein delivery vectors. AB - The development of novel cancer therapies that are selective for cancer cells with limited toxicity to normal tissues is a challenge for oncology researchers. Microorganisms, such as viruses with selectivity for tumor cells or tumor micro environments, have been investigated as potential arsenals for decades. Genetically-modified, non-pathogenic bacteria have begun to emerge as potential antitumor agents, either to provide direct tumoricidal effects or to deliver tumoricidal molecules. Attenuated Salmonella, Clostridium and Bifidobacterium are capable of multiplying selectively in tumors and inhibiting their growth, representing a new approach for cancer treatment. Because of their selectivity for tumor tissues, these bacteria would also be ideal vectors for delivering therapeutic proteins to tumors. VNP20009, an attenuated strain of Salmonella typhimurium, and its derivative, TAPET-CD, which expresses an Escherichia coli cytosine deaminase (CD), are particularly promising, and are currently undergoing phase I clinical trials in cancer patients. PMID- 11926127 TI - Recombinant antibodies for the selective targeting of tumor neovasculature. AB - Angiogenesis, the sprouting of new blood vessels from preexisting ones, is a characteristic feature of many aggressive solid tumors. Advances in the identification of markers of angiogenesis and new methodologies for the production of human recombinant antibodies are making it possible to deliver bioactive molecules to the tumor neovasculature in a selective fashion. This review illustrates how recombinant antibody derivatives may open new avenues for the imaging and therapy of angiogenesis-related diseases, such as cancer, blinding ocular disorders and rheumatoid arthritis. PMID- 11926128 TI - Inhibitors of growth factor receptor kinase-dependent signaling pathways in anticancer chemotherapy--clinical progress. AB - Aberrant signal transduction plays a major role in the pathophysiology of cancer. Kinases, key enzymes involved in signaling pathways, are attractive targets for chemotherapeutic intervention in the fight against cancer. Many cancers respond to endogenous growth factors or, through autocrine loops, express growth factors and the requisite receptor kinases, resulting in cellular proliferation. Growth factor-induced signaling has also been implicated in the activation of anti apoptotic cell survival pathways. This review attempts to describe some of the key advances made in the last year towards the design and development of inhibitors of growth factor receptor kinases. Particular emphasis is placed on ATP-competitive, small molecule inhibitors of this important class of enzymes. PMID- 11926129 TI - Current progress on farnesyl protein transferase inhibitors. AB - The hypothesis that the inhibition of farnesyl protein transferase (FPTase) may be beneficial in the treatment of certain cancers is finally being clinically tested. The efforts of almost a decade of preclinical research have resulted in several new chemical entities that are currently undergoing phase I, II or III clinical trials. The number of compounds being tested is small and disproportionate given the amount of effort by a large number of organizations in targeting FPTase. The three compounds, BMS-214662 (Bristol-Myers Squibb), R 115777 (Janssen Pharmaceutica BV) and Sch-66336 (Schering-Plough Research Institute) are the most advanced. While initial results with these three candidate drugs are encouraging, it is disappointing that the clinical efficacy of many other candidates has been less successful. In this review, we summarize the clinical and preclinical data published in 2001, and discuss the natural product inhibitors reported between 1998 to 2001. In addition, a comprehensive history of the FPTase inhibitory discovery effort at Merck, from CAAX tetrapeptides to peptidomimetics to macrocyclic compounds, is summarized for the first time. PMID- 11926130 TI - Recent developments in peptide-based cancer therapeutics. AB - Peptides make up only a fraction of therapeutically beneficial pharmaceutical molecules that are either in clinical testing or in the market. Despite the initially low number and impact of peptides on therapeutics, their potential has just intensified with new developments in modifications, stability, delivery and preclinical success. With the completion of the human genome sequence and developments in the proteomics field, peptides are emerging as important molecules for cancer therapy. Several peptides with exciting preclinical results have now entered into clinical trial for the treatment of human cancers. In this review, some of the recent advances in peptide-based cancer therapeutics will be discussed. PMID- 11926131 TI - The cyclotides: novel macrocyclic peptides as scaffolds in drug design. AB - Cyclotides are a novel class of circular, disulfide-rich peptides (approximately 30 amino acids) that display a broad range of bioactivities and have exceptionally high stability. Their physical properties, which include resistance to thermal and enzymatic degradation, can be attributed to their unique cyclic backbone and knotted arrangement of disulfide bonds. The applicability of linear peptides as drugs is potentially limited by their susceptibility to proteolytic cleavage and poor bioavailability. Such limitations may be overcome by using the cyclotide framework as a scaffold onto which new activities may be engineered. The potential use of cyclotides for drug design is evaluated here, with reference to rapidly increasing knowledge of natural cyclotides and the emergence of new techniques in peptide engineering. PMID- 11926132 TI - Therapeutic applications of guanylate cyclase-C receptor agonists. AB - Uroguanylin, guanylin, lymphoguanylin and bacterial enterotoxin ST are structurally related peptides that activate common guanylate cyclase signaling molecules and, via cyclic GMP (cGMP), regulate water and ion homeostasis in a variety of tissues and organs, including the gastrointestinal (GI) tract, kidneys, lung, prostate, etc. Any disruptions in water and ionic homeostasis due to reduced production of one or more of the guanylin family of peptides can potentially lead to carcinogenic and inflammatory stimuli. Recent findings that expression of uroguanylin and guanylin is dramatically reduced in polyps and colon carcinoma tissues further support the notion that these peptide hormones are important modulators or mediators of signaling pathways leading to the processes involved in carcinogenesis and inflammatory disorders. This review briefly discusses recent research findings suggesting the novel therapeutic applications of agonist peptides of guanylate cyclase-C receptor. PMID- 11926133 TI - Bacterial toxins as versatile delivery vehicles. AB - An ability to deliver macromolecules into the intracellular compartments of mammalian cells offers enormous potential for development of new therapeutics directed against intracellular targets. Unfortunately, most peptides or proteins are too large to enter the cell cytosol unaided, and any uptake that does occur primarily results in their entry into lysosomes for degradation. However, one group of proteins that possesses an inherent capacity to interact with and enter mammalian cells are bacterial toxins. These are being developed as efficient vehicles for the attachment and intracellular delivery of other macromolecules, including peptides, proteins and DNA. To date, most studies have concentrated on the delivery of immunological epitopes into the endogenous major histocompatibility class I (MHC-I) pathway for development of antiviral or anticancer vaccines. However, opportunities to use toxins to modulate inflammatory autoimmune disorders and cell-specific targeting of DNA for gene therapy illustrates the versatility of toxin molecules as delivery vehicles. PMID- 11926134 TI - Drug delivery systems based on sugar-macromolecule conjugates. AB - The specificity of carbohydrate-protein interactions can greatly outstrip that of many other ligand-binding systems; such is the enormous density of information that sugars can convey. In addition, macromolecules allow for the fine-tuning of active drug delivery through their great ability to undergo site-specific modification and their inherent physicochemical properties. Once combined, these two factors suggest that sugar-macromolecule conjugates, targeted using endogenous carbohydrate binding proteins, are a promising route to the 'magic bullet'. PMID- 11926135 TI - Use of HSV vectors to modify the nervous system. AB - Herpes simplex virus (HSV) is a natural human pathogen that efficiently infects sensory neurons to establish a life-long latent state. Recombinant replication defective vectors, created by disruption of critical viral gene functions, nonetheless target neurons and can be used to express transgenes to alter the structure and/or function of the nervous system. Specific applications of these vectors to models of neurodegeneration (Parkinson's disease), trauma (spinal root avulsion), peripheral neuropathy and neuronal function (pain) have been published within the last year. With these applications and the clinical experience in human tumor trials with HSV vectors, the stage is set for the use of HSV-based vectors to treat neurologic disease in humans in the near future. PMID- 11926136 TI - Plant cell cultures as a source of bioactive small molecules. AB - Plants have provided some of our most important pharmaceuticals and are a continuing source of novel bioactive molecules for drug development. Cell suspension cultures of higher plants are a complementary source of novel chemistry with significant advantages over whole plants, including plasticity of expression and re-accessibility. With an emphasis on recent progress, the use of plant cell cultures for both new lead discovery and as a route to the production of known bioactives will be described. Developments in strategies that can be applied to facilitate future success in the area are discussed. PMID- 11926137 TI - Mother nature's combinatorial libraries; their influence on the synthesis of drugs. AB - Natural products or secondary metabolites, whether from the microbial, plant or marine worlds, represent the results of evolutionary pressures to preserve and enhance the life of their producing organism. They have evolved into structurally and usually stereochemically complex compounds with specific bioactivities. They thus represent a diverse 'combinatorial library' that may have potential pharmaceutical use. In principle, the combination of this diverse library with the methods of combinatorial chemistry could lead to an unlimited supply of diverse and complex structures, and is recommended as a fruitful approach for future drug development. Examples of the application of combinatorial methods to nature's combinatorial library will be presented and discussed, with an emphasis on the antitumor, anti-infective and pain control disease areas. PMID- 11926138 TI - [Insulin sensitivity measured with euglycaemic clamp technique in newly diagnosed diabetes mellitus]. AB - The purpose of the study was to identify insulin resistance (IR) at the onset of diabetes mellitus type 2 and type 1 using euglycaemic clamp technique. The study was conducted in 51 subjects: 30 patients with newly diagnosed diabetes, 9 with type 1, mean age 29.3 +/- 6.6 yrs (SD), 21 with type 2, mean age 47.8 +/- 10.7 yrs and 21 control healthy subjects (mean age 43.5 +/- 13.9 yrs). In last group there was an increase in IR proportional to patients age (r = 0.78, p < 0.01). IR was more pronounced in obese subjects in comparison to non-obese subjects, index of tissue glucose disposal M was respectively 3.4 +/- 2.2 and 6.6 +/- 1.7 mg/kg x min, p < 0.005. M value was decreased also in patients with abdominal obesity measured with WHR, r = 0.73, p < 0.0005. The normal range for this method was 3.3 to 10 mg/kg x min. Insulin sensitivity is decreased in both types of newly diagnosed diabetes, in type 1 to 2.3 +/- 0.5 and in type 2 to 1.7 +/- 0.8 mg/kg x min as compared with control group. In diabetes type 1 IR was correlated with degree of hyperglycaemia measured with HbA1c ("glucotoxicity"). The lowest M values was found in diabetes type 2 with obesity. There were no significant abnormalities in insulin secretion in diabetic groups. These results suggest that the primary disturbance at the onset of diabetes mellitus is IR. PMID- 11926139 TI - [Plasma concentration of leptin, neuropeptide Y and tumor necrosis factor alpha in patients with cancers, before and after radio- and chemotherapy]. AB - In patients with cancers progressive reduction of body mass is frequently recent. Pathogenesis of cachexia in patients with cancer is multifactorial. Such factors as cytokines, peptides relieved by tumor mass and different forms of treatment as radio or chemotherapy may play a major role in the pathogenesis of cachexia in patients with cancer. The aim of this study was to assess the relationship between body fat and lean mass and plasma leptin, NPY and TNF concentrations in patients with cancer of oral cavity and pharynx, cancer of larynx and non-Hodgkin lymphoma (NIL). 30 patients (10 with cancer of oral cavity and pharynx, 10 with cancer of larynx and 10 with non-Hodgkin lymphoma) were enrolled into this study. Mean age of all cancer patients was 50 +/- 2.9 years (from 18 to 76 years). The control group consisted of 29 healthy subjects with a means age 48 +/- 3.5 years (from 18 to 75 years), properly chosen according to the body weight, BMI, gender and age as above mentioned groups of patients with cancer. In control and study groups body fat and not-fat mass was assessed before and after treatment using the bioelectrical impedance method. Before oncological therapy patients with cancer did not differ from healthy subject with regard to body weight and body mass index. After treatment significant: decrease of body weight, body fat mass and BMI was observed. Serum leptin, NPY and TNF concentrations were analysed in healthy subjects and patients with cancer before and after treatment. Before oncological treatment significantly lower serum leptin concentration in comparison to leptinaemia in control group was found. In contrast to serum leptin, NPY serum concentration was similar in patients with cancer and in control subjects. Serum concentration of TNF was significantly higher in patients with cancer in comparison to subjects of control group. After oncological treatment, serum leptin and NPY concentration did not change significantly. In contrast, serum TNF concentration decreased significantly after oncological therapy. From the results obtained in this study we can conclude, that in patients with cancer secretion of leptin is decreased in relation to body fat mass. However, contribution of this hormone to pathogenesis of cancer induced anorexia seems not to proven. From the other side, the role of TNF in pathogenesis of disregulation of leptin secretion seems to be very likely. After chemo or radiotherapy, serum NPY concentration did not change significantly. After this oncological treatment the relationship between serum leptin concentration and body mass is no longer significant. PMID- 11926140 TI - [Left and right ventricular function in acromegalic patients]. AB - Recent studies have proved close relations between cardiovascular and endocrinic systems. This relation has been observed in acromegaly, the disease connected with unrestrained secretion of growth hormone. The aim of the study was to assess Holter monitoring and echocardiography of acromegalic patients. The study group consisted of 28 acromegalic patients, including 15 patients with hypertension, was considered. As control groups we examined 20 patients with essential hypertension and 20 normotensive healthy subjects: All subjects underwent twenty four hour Holter recordings, complete M-mode, two-dimentional and spectral Doppler echocardiography. Ventricular premature complexes occurred in 65% of acromegalic patients. Frequency and severity of ectopic beats were significantly increased compared to control groups. Left ventricular ejection fraction was considered to be normal, although significant decreased compared to healthy subjects. Left ventricular mass was above normal value in acromegalic patients- no significant difference was found between hypertensive and normotensive acromegalics. Doppler examination has shown the abnormalities of left and right ventricular filling in 89% of acromegalics. We have observed the correlations between left and right ventricular filling indices and the duration of the disease, and left ventricular mass. Left ventricular hypertrophy frequently occurs in acromegalic patients and this is not simply secondary to systemic hypertension. The prevalence of ventricular arrhythmias in acromegalic patients seems to be associated with left ventricular hypertrophy. Doppler examination suggests impaired left and right diastolic filling in patients with acromegaly. PMID- 11926141 TI - [Radiofrequency ablation of accessory pathway's atrial insertion site]. AB - Brought into clinical practice in 80, method of radiofrequency catheter ablation (RFCA) enabled possibility of causative treatment in patients with preexcitation syndrome. RFCA may be performed utilizing various techniques, choice of one of them depends on experience and preference of the operator. Aim of the study was assessment of efficacy and safety of RFCA targeting accessory pathway's atrial insertion site with help of unipolar signals from ablation-catheter with regard to various localizations of accessory pathways. Material and method studied population consisted of consecutive 149 patients with symptomatic, drug-resistant WPW-syndrome, which were admitted to our Center for electrophysiologic study and RFCA. From the studied population two groups were subdivided: patients with left sided (group I, n = 112) and right-sided (group II, n = 37) accessory pathway. Every patient underwent electrophysiological study with use of four diagnostic electrodes and after preexcitation was confirmed RFCA was undertaken. In case of left-sided accessory pathway transseptal puncture was performed. Analyzed parameters were: total duration, fluoroscopy-time, number of current-applications during RFCA, as well as success-rate, complications and recurrences after successful ablation. Patient in both groups did not differ significantly with respect to age and sex. Success-rate was insignificantly higher in gr.I (92.8%) than in gr.II (86.5%), as was RFCA-duration (136.4 min versus 123.6 min). We found significant differences in fluoroscopy-time, which was longer in gr.I (37.2 min versus 28.4 min, p < 0.05). Complications occurred in 1 patient in each group. RFCA aiming at accessory pathway's atrial insertion site with use of unipolar tracings from ablation-catheter is safe and effective method of treatment in WPW-patients. PMID- 11926142 TI - [Uniform programme in caring for patients with acute hepatic porphyria]. AB - The programme commissioned by the Ministry of Health is presented. Data basis for the years 1996-1998 were created. A booklet on diagnosis, treatment and prophylaxis of porphyrias was edited and distributed in Poland in 5000 copies to all hospitals, major laboratories, medical libraries, regional officers as well as patients. The booklet lists simple diagnostic methods which can be easily introduced to hospital laboratories for early diagnosis of porphyria. It also includes a list of the safe drugs for patients with porphyria. Several lectures for doctors and medical staff were delivered, nine publications appeared and six others were accepted for publication. Results of the work, evaluated after three years are as follows. Early clinical diagnosis of porphyria and its prophylaxis among family members has significantly improved. The frequency of severe attacks and the mortality rate due to porphyria has been reduced to singular cases. The proposed organization of small regional centers (with help of Porphyria Center), oriented at early diagnosis of porphyria appears to be impossible without further financial support. PMID- 11926143 TI - [Effect of postinfarct left ventricular dysfunction on late occurrence of mitral regurgitation--two year experience]. AB - The aim of this study was to evaluate factors that might influence on the occurrence of mitral regurgitation (MR) in patients (pts) following myocardial infarction (MI) after fibrinolysis--two year experience. The study group comprised 118 pts: 40 women and 78 men (mean age: 58 +/- 9 years) following MI, who underwent Doppler echocardiography which revealed no MR 3 weeks after the MI. The second echocardiographic examination was performed after 2 years. We evaluated the following parameters: presence and stage of MR, left ventricular end-diastolic diameter (LVDD), left atrial diameter (LA), end-diastolic volume (EDV), wall motion score index (WMSI), asynergic area (AA) and ejection fraction (EF). Results after 2 years were as follows: 43 pts (36%) presented without MR, 45 (38%) MR I degree, 27 (23%) MR II degree, 3 (3%) MR III degree. Echocardiographic parameter comparison after 2 years demonstrated the following changes: LVDD increase from 5.0 +/- 0.5 cm to 5.3 +/- 4.6 cm (p < 0.005), LA increase from 3.7 +/- 0.4 cm to 4.3 +/- 0.4 cm (p < 0.00001), EDV increase from 130 +/- 29 ml to 147 +/- 39 ml (p < 0.005), WMSI increase from 1.37 +/- 0.23 to 1.45 +/- 0.21 (p < 0.05), AA increase from 23.5 +/- 10.1% to 27.8 +/- 7.9% (p < 0.005) and significant EF decrease 50.4 +/- 7.9% to 46.9 +/- 7.1% (p < 0.005). These results demonstrate that the occurrence of MR 2 years after the MI is caused by left ventricular remodelling, as well as segmental and global function deterioration. PMID- 11926144 TI - [Estimation of efficacy of the octreotide LAR administration in the patients with somatotropinoma]. AB - Acromegaly is caused by excessive secretion of growth hormone by a hypophyseal adenoma type of somatotropinoma. IGF-I is formed in the liver and mediates most biological actions of GH. Treatment of adenomas, which secrete GH, involves pharmacotherapy followed by surgery. Modern pharmacotherapy leaning is based on somatostatin analogues (factor restrictive secretion GH): octreotide, octreotide LAR and lanreotide. The aim of our study was estimation of efficiency of octreotide LAR in the patients with somatotropinoma prepared to neurosurgery intervention. We examined 16 patients (10 of women and 6 men) with the features of active acromegaly. In all cases the increased concentration of HGH and IGF-I were observed. The presence of pituitary adenoma in all patients was confirmed by MRI. The patients were treated with octreotide LAR monthly in dose 20 mg and 30 mg respectively. Before and after application of somatostatin analogues the concentration HGH, IGF-I, PRL in serum were marked. The concentration of GH before octreotide LAR therapy in all patients increased remarkable and ranged from 15.6 to 78.6 ng/ml, mean: 31.20 +/- 16.84 (norm: 0-10 ng/ml), also, in all cases the serum IGF-I level was increased and ranged from 451 to 1107.6 ng/ml, mean: 801.75 +/- 207.82 (norm: 100-400 ng/ml). The prolactin concentration ranged from 7.4 to 49.9 ng/ml, mean: 22.8 +/- 13.7 (norm: 2-20 ng/ml) and in 8 (50%) cases the increased of PRL concentration in serum was observed. After the administration of octreotide LAR the level of: GH [mean: 12.99 +/- 17.16 ng/ml (p < 0.001)], of IGF-I [mean 422.8 +/- 229 ng ml (p < 0.01)] statistical important decreased and prolactin in 8 with increased concentration [mean: 12.45 +/- 5.57 (p < 0.01)] were observed. Long acting somatostatin analogues--octreotide LAR is particular efficient in lowering of growth hormone and IGF-I in patients with somatotropinoma and shows efficiency in normalization of increased prolactin concentration. Because of extreme effectiveness of octreotide LAR, it should be used the routine treatment at the patients suffering from active acromegaly and preparing to neurosurgical treatment. PMID- 11926145 TI - [Myocardial infarction in young female smoker taking oral contraceptives]. AB - A 41-year-old woman, heavy smoker, was admitted to the hospital post an apical myocardial infarction, which occurred 16 days before. Chest pain appeared two weeks before the infarction. There were no other cardiovascular risk factors except heavy prolonged nicotine use. She has started oral contraceptives treatment one month before MI. The patient underwent coronary angiography, which did not show any abnormalities. Left ventriculogram confirmed the apical localization of infarction. The most probable pathomechanism of MI was a coincidence of oral contraceptives use and smoking. PMID- 11926147 TI - [The role of immune mechanisms in the pathogenesis of atherosclerosis]. PMID- 11926146 TI - [Liver sarcoidosis--a case report]. AB - 46-years old woman was diagnosed in Department of Infectious Diseases of Medical Academy in Wroclaw due to dyspeptic disorder, eosinophilia and high AspAT, AlAT, AP and GGTP level in blood serum. Liver biopsy was performed. Hepatic, noncaseating granulomata were found in liver biopsy specimen. Histological changes indicate to liver sarcoidosis. PMID- 11926148 TI - [Androgen therapy in the management of osteoporosis in men without distinct hypotestosteronemia]. PMID- 11926149 TI - [Detectable plasma markers of atherosclerotic artery injuries]. PMID- 11926150 TI - [Bisphosphonates in the treatment of breast carcinoma]. PMID- 11926151 TI - The eyes have it! PMID- 11926152 TI - Eye signs in systemic disease. AB - BACKGROUND: The eye provides clues to the diagnosis of many systemic diseases and many important complications of these diseases occur in the eye. OBJECTIVE: To review the important ocular pathology seen in some systemic diseases. The ocular manifestations of a small number of these diseases will be discussed in detail. DISCUSSION: Examination of the eye should be a routine and important part of a general examination in certain systemic diseases. PMID- 11926153 TI - Sticky eye, tricky diagnosis. AB - BACKGROUND: Infective conjunctivitis is common and mostly responds well to supportive management and/or empiric antibiotic treatment. At times the differential diagnosis includes more serious conditions that potentially threaten vision. Recurrence and treatment failures are relatively common. OBJECTIVE: To outline the usual clinical appearance of conjunctivitis and its management. To explore the clinical appearance (with the help of case studies) of conditions that have been mistaken for conjunctivitis, and to consider some of the conditions that may coexist that contribute to recurrence and nonresponsiveness. DISCUSSION: Empiric treatment for presumed bacterial conjunctivitis remains an appropriate course of action. Care must be taken in certain groups who are at high risk of complications. Careful review of the clinical appearance and consideration of other possible diagnoses is mandatory in nonresponding patients. PMID- 11926154 TI - Emergency primary eye care. Tips for diagnosis and acute management. AB - BACKGROUND: Acute eye problems are among the commonest problems presenting to the general practitioner. Due to the general lack of exposure to such problems and unavailability of appropriate equipment, the diagnosis is difficult to make with any degree of certainty. The availability of eye services varies widely in Australia. In urban centres the GP has greater access to ophthalmic services. In regional Australia, the practitioner may need to rely more on his or her own findings and experiences. OBJECTIVE: This article aims to equip the GP with an outline of the clinical findings as well as an approach to acute management of the commonest presenting problems. In many cases, initial treatment will serve to minimise morbidity, before treatment can be delivered by an ophthalmologist. DISCUSSION: I have divided the commonest problems that may need urgent attention into three groups: acute red eye; trauma; and sudden visual loss. This list is not intended as an exhaustive presentation of ophthalmic problems, but rather an outline of conditions where prompt diagnosis and treatment by the GP can positively affect the outcome. PMID- 11926155 TI - Surgical options for eyelid problems. AB - BACKGROUND: Eyelid problems may be vision threatening, painful, irritating, cause watering, or just unsightly. Many are treatable surgically. OBJECTIVE: This article describes some of the more common eyelid conditions, their presentation, investigation and surgical treatment. DISCUSSION: Common eyelid conditions include ectropion, entropion, ptosis, thyroid eye disease and facial palsy. Treatments, including surgical options, are discussed. Most eyelid surgery on adults can be done as day surgery under local anaesthetic with intravenous sedation. PMID- 11926156 TI - Unresolved lateral ankle pain. It's not always 'just a sprain'. AB - BACKGROUND: Lateral ankle sprains are the most common sports related injury. Unfortunately a number of other lateral ankle injuries are often misdiagnosed as a sprain, resulting in misdirected therapy, delayed functional return and unresolved ankle pain. OBJECTIVE: This review focuses on the differential diagnosis and management of unresolved lateral ankle pain, with an emphasis on methods to correctly identify the injury. DISCUSSION: A surprisingly long list of conditions can mimic a lateral ankle sprain, including syndesmosis sprains, subtalar sprains with or without instability, impingement lesions, chronic ankle instability, osteochondral lesions of the talus, peroneal tendon injury, tarsal coalition and fractures of the hindfoot. A careful history, anatomically accurate palpation and plain erect X-rays are the cornerstone of diagnosis. The key is to remember that it's not always just an ankle sprain. Bone scintigraphy, CT and MRI are valuable investigational adjuncts. Arthroscopy is an excellent method of intra-articular visualisation to diagnose and treat unresolved lateral ankle pain. PMID- 11926157 TI - Post-kernicteric syndrome. AB - Traditionally, neonates in Australia received specialised care in hospitals for about a week, by which time any developing jaundice would have been detected and managed. However, with the introduction and popularity of early maternal discharge about 48 hours after delivery (in Australia and elsewhere), the duty of diagnosing neonatal jaundice and referring newborns for urgent management falls on general practitioners. Delays or nonreferrals predispose infants to kernicterus or bilirubin encephalopathy, a previously uncommon cause of brain damage. In the interest of patient safety, family physicians need clinical guidelines for managing jaundice, as kernicterus is a preventable tragedy--and increasingly reported in journals. Until guidelines are available, hopefully this cautionary tale will assist GPs in the risk management of all jaundiced infants. PMID- 11926158 TI - One pair must last a lifetime. Foot care and diabetes. AB - This is the first in a series of six articles on foot care, covering diabetes, vascular disease, neuropathy, painless ulcers and structural abnormalities. The management strategies will help patients 'make one pair last a lifetime'. PMID- 11926159 TI - Asylum seeking and mandatory detention. The psychological consequences. PMID- 11926160 TI - Doctor as patient. Part 3: trying to resume normal life. PMID- 11926161 TI - Changing clinician practice. The RACGP/RACO National GP Eye Skills Workshop. AB - AIM: To evaluate the impact of the Royal Australian College of General Practitioners (RACGP)/Royal Australian College of Ophthalmologists National General Practitioner Eye Skills Workshop on Queensland participants reported medium and long term ocular practice pattern. METHOD: Two questionnaires were administered to all Queensland workshop participants (n = 121), six months and > 12 months following the workshop, to assess self reported ocular practice pattern. RESULTS: The response rate was identical for both the first and second questionnaire with 78 (64%) GPs returning completed forms on each occasion. Results indicate a highly significant increase in the proportion of GPs having access to dilating drops in the practice, using a systematic approach to eye assessment, and using the ocular kinetic perimetry (OKP) chart (p < 0.001), performing paediatric vision testing (p = 0.002) and incising chalazion (p = 0.01) following the workshop. General practitioners also reported a significant increase in the use of dilatation and fundoscopy in all clinical areas surveyed (p < 0.001). The change in dilatation and fundoscopy practice found at six months was sustained at > 12 months. CONCLUSION: Our study has demonstrated the National GP Eye Skills Workshop to be associated with significant and sustained practice change in key areas of ocular assessment in general practice. A key ingredient in the workshop's success was the focus on experiential learning, intra-session practice rehearsal and use of local opinion leaders. PMID- 11926162 TI - Osteoporosis risks. A comparative study of Asian Australian and Caucasian Australian women. AB - OBJECTIVE: To describe the self reported level of osteoporosis knowledge, attitudes and preventive activities, including dietary calcium intake of Asian and Caucasian women. METHOD: Self administered questionnaires in English and Chinese including a food frequency survey were posted to potential respondents that were recruited via general practices, community newsletters and notices. RESULTS: Asian women had less osteoporosis knowledge and lower calcium intake (mean 793 +/- 49 mg/day standard error of the mean) than Caucasian women (1298 +/ 56 mg/day). Postmenopausal Asian women were at greater risk of being below the recommended daily intake (73%) than Caucasian women (31%). CONCLUSION: Osteoporosis knowledge and preventive activities among Asian and Caucasian women could be better. Higher dietary or supplementary calcium intake should be considered. PMID- 11926163 TI - Needle length required for intramuscular vaccination of infants and toddlers. An ultrasonographic study. AB - BACKGROUND: To be maximally effective and to induce less adverse reactions, injection with many vaccine antigens must penetrate muscle rather than subcutaneous tissue. AIM: To determine the length of needle needed to penetrate muscle at the anterolateral thigh vaccination site in children aged two, four, six and 18 months. METHOD: Ultrasound measurements were made of the subcutaneous and muscle layer thickness of children aged two, four, six and 18 months at the junction of the upper and middle thirds of the anterolateral thigh with the probe applied parallel to the long axis of the leg and at 45 degrees to the vertical and at 90 degrees to the skin's plane. RESULTS: Subcutaneous tissue (SCT) and muscle layer (ML) thickness were measured in 57 children (2 months, n = 14; 4 months, n = 13; 6 months, n = 18; 18 months, n = 12) with mean SCT thickness of: 8.6 +/- 3.0 mm at 2 months; 9.4 +/- 2.0 mm at 4 months; 10.2 +/- 2.1 mm at 6 months; and 8.1 +/- 1.7 mm at 18 months. Muscle layer thickness in these children was: 10.5 +/- 2.4 mm at 2 months; 12.2 +/- 2.0 mm at 4 months; 14.8 +/- 2.0 mm at 6 months, and 16.5 +/- 4.6 mm at 18 months. CONCLUSION: The optimal needle length to routinely penetrate muscle of the anterolateral thigh in children aged two, four, six and 18 months depends on the technique employed. A 16 mm long needle is suitable with the WHO technique (injecting at 90 degrees to skin's surface) and 25 mm long needle with the NH&MRC and American techniques (injecting at 45 degrees to skin's surface). PMID- 11926164 TI - A questionnaire to measure general practitioners' attitudes to their role in the management of patients with depression and anxiety. AB - OBJECTIVE: Attitude, together with knowledge, is important in determining behaviour. Doctors' attitudes affect the recognition and management of mental health. We detail the development of an instrument to measure general practitioner attitudes to the recognition and management of depression and anxiety disorders. METHOD: A literature review and semistructured interviews with GPs identified constructs for inclusion into the questionnaire. Items were written and examined by a panel of psychiatrists, GPs and statisticians to establish face and content validity. The resulting 30 item questionnaire was given to a convenience sample of 63 GPs in Australia with an interest in mental health, and their responses analysed using exploratory factor analysis. RESULTS: Exploration of the data identified two factors. Factor 1 contained 12 items measuring professional comfort and competence with the care of mental health disorders (alpha = 0.82). Factor 2 contained five items that assessed GP concerns about difficulties with the health care system in this area (alpha = 0.73). DISCUSSION: The factor analysis shows that GPs' attitudes can be parsimoniously described by two factors, instead of the five that the literature suggests. The high values of Cronbach's alpha demonstrate that these two factors can be measured reliably. These were not the anticipated results, and can be taken to indicate that GPs' attitudes to their roles in the recognition and management of patients with depression and anxiety disorders are less complex than might otherwise be imagined. PMID- 11926165 TI - Effective immunotherapy of cancer in MUC1-transgenic mice using clonal cytotoxic T lymphocytes directed against an immunodominant MUC1 epitope. AB - The tumor-associated autoantigen MUCI is intensively studied as a potential target for antigen-specific immunotherapy of cancer. Previous reports concerning experiments in preclinical murine tumor models have provided evidence supporting the feasibility of this approach. However, such studies have not been performed with clonal cytotoxic T lymphocyte populations displaying a highly defined MUC1 specificity. The authors demonstrate that the immunodominant MUC1-specific cytotoxic T lymphocyte response in C57BL/6 mice is directed against an H-2Kb restricted epitope, MUC1(19-27), which is derived from the N-terminal signal sequence of the MUC1 protein. Processing of this epitope was independent of transporter of antigen presentation and proteasome function. Importantly, successful immunotherapy of MUC1-overexpressing tumors in MUC1-transgenic mice was not accompanied by damage to normal somatic MUC1-positive tissues, even when this involved the infusion of large numbers of clonal cytotoxic T lymphocyte that recognized the immunodominant MUC1 epitope. Although the risk for autoimmune pathology is limited, data indicate that immune tolerance in MUC1-positive subjects restricts the breadth of the MUC1-specific cytotoxic T lymphocyte repertoire that is available for recruitment to immunotherapeutic antitumor responses. PMID- 11926166 TI - Threat-induced cortical processing and startle potentiation. AB - This paper presents cortical responses as reflected in event-related potentials (ERP) in an instructed fear paradigm. Safe cues and threat cues that predict shock were presented at an unprecedented fast rate (mean SOA of 2.1 s). Startle and subjective measures confirmed that threat relative to safe cues elicited fear. Several ERP correlates of fear processing were predicted and confirmed: modulation of exogenous sensory components, frontal selection positivity, and increase of P3. Furthermore, a frontal negative slow wave was observed. These results are discussed in relation to attentional selection models and emotional processing. PMID- 11926167 TI - Epidural cooling for the prevention of ischemic injury to the spinal cord during aortic occlusion in a rabbit model: determination of the optimal temperature. AB - PURPOSE: This experiment was designed for the determination of the optimal epidural cooling temperature for the allowance of spinal cord protection with minimal side effects during an aortic occlusion-induced spinal cord ischemia model in rabbits. METHODS: Spinal cord ischemia was induced in rabbits with infrarenal aortic occlusion for 40 minutes. Spinal cord cooling was effected with epidural infusion of normal saline solution at the following different temperatures: group 1, 17 degrees C (n = 6); group 2, 24 degrees C (n = 6); group 3, 32 degrees C (n = 6); and group 4, 39 degrees C (n = 3). Sham-operated rabbits without aortic occlusion were used as controls with epidural infusion at healthy body temperature (39 degrees C; n = 3). Motor function was assessed at 48 hours with Tarlov's criteria, and the animals were killed. The spinal cord was sectioned into multiple segments, and semiquantitative histologic scoring (0 to 5) was used to grade ischemic injury. RESULTS: Cooling solution and spinal cord temperatures showed linear correlation (r = 0.95). All the rabbits in groups 1 (except one with mild weakness), 2, and 3 were neurologically intact, and all in group 4 had paraplegia develop (P < .001). One rabbit in group 1 died from increased intracranial pressure (ICP). Mean blood pressure, ICP, and body temperature were similar among the groups. Histology correlated with the clinical findings. In groups 1 and 2, minimal histologic changes were noted. Low-grade ischemic changes were present in group 3 in the low-lumbar and mid-lumbar segments. Severe ischemic injury occurred at the same segments in group 4 (P < .05). CONCLUSION: These study results suggest that in rabbits satisfactory spinal cord protection during aortic occlusion can be achieved at moderate regional hypothermia (24 degrees C). Large volume infusion for the achievement of profound hypothermia may cause deleterious effects of increased ICP and is not warranted. PMID- 11926168 TI - "We are under attack". PMID- 11926170 TI - Differential susceptibility to ageing of rat preganglionic neurones projecting to the major pelvic ganglion and of their afferent inputs. AB - We have analysed age-related changes in the morphology of preganglionic neurones in the lumbosacral spinal cord, labelled following injection of retrograde tracers into the major pelvic ganglion of young adult and aged male rats. We have also examined changes in neurotransmitter-characterised spinal afferent inputs to these neurones, or to the nuclei in which they lie, using light and electron microscope immunohistochemistry. In previous investigations of the major pelvic ganglion, the sympathetic, but not parasympathetic, postganglionic neurones were seen to exhibit age-related changes and the same pattern is seen in the preganglionic neurones. This included an apparent reduction in the numbers of sympathetic preganglionic neurones, and a reduction in the length of their dendrites and the complexity of their branches. Ultrastructural immunohistochemical studies described here reveal significant reductions in the area of synaptic contact made by glutamate-immunoreactive boutons onto the dendrites of sympathetic (but not parasympathetic) preganglionic neurones, while contacts from boutons immunoreactive for glycine or gamma-aminobutyric acid (GABA) were unchanged. There is also a reduction in synaptic contacts received by sympathetic somata from boutons immunoreactive for none of these amino acids. Serotonin-immunoreactive terminals are closely associated with preganglionic autonomic neurones, and these are reduced in number in sympathetic, but not parasympathetic, spinal nuclei of aged rats. However, serial section electron microscopy has so far failed to demonstrate conventional synaptic contacts between serotonergic terminals and the dendrites or somata of the preganglionic autonomic neurones. In young animals, axon terminals immunoreactive for thyrotropin-releasing hormone (TRH) are abundant in all spinal laminae including area X, but in aged animals, such terminals are significantly reduced in number in regions containing preganglionic sympathetic, but not parasympathetic, neurones. These results indicate that the sympathetic preganglionic neuron populations that project to the major pelvic ganglion, and the spinal inputs they receive, show a number of degenerative changes in aged rats which are not seen parasympathetic preganglionic neuronal populations. PMID- 11926172 TI - Peptide nucleic acid probe detection of mutations in Mycobacterium tuberculosis genes associated with drug resistance. AB - The emergence of drug-resistant strains of Mycobacterium tuberculosis is a serious public health problem. Many of the specific gene mutations that cause drug resistance in M. tuberculosis are point mutations. We are developing a PCR peptide nucleic acid (PNA)-based ELISA as a diagnostic method to recognize point mutations in genes associated with isoniazid and rifampin resistance in M. tuberculosis. Specific point mutation-containing sequences and wild-type sequences of cloned mycobacterial genes were PCR-amplified, denatured, and hybridized with PNA probes bound to microplate wells. Using 15-base PNA probes, we established the hybridization temperatures (50 degrees C-55 degrees C) and other experimental conditions suitable for detecting clinically relevant point mutations in the katG and rpoB genes. Hybridization of PCR-amplified sequences that contained these point mutations with complementary mutation-specific PNAs resulted in significant increases in ELISA response compared with hybridization using wild-type-specific PNAs. Conversely, PCR-amplified wild-type sequences hybridized much more efficiently with wild-type PNAs than with the mutation specific PNAs. Using the M. tuberculosis cloned genes and PCR-PNA-ELISA format developed here, M. tuberculosis sequences containing point mutations associated with drug resistance can be identified in less than 24 h. PMID- 11926171 TI - Calmodulin-dependent cyclic nucleotide phosphodiesterase in an experimental rat model of cardiac ischemia-reperfusion. AB - In the present study, we investigated the activity and expression of calmodulin dependent cyclic nucleotide phosphodiesterase (CaMPDE) and the effects of calpains in rat heart after ischemia and reperfusion. Immunohistochemical studies indicated that CaMPDE in normal heart is localized in myocardial cells. Rat ischemic heart showed a decrease in CaMPDE activity in the presence of Ca2+ and calmodulin; however, in ischemic-reperfusion tissue a progressive increase in Ca2+ and calmodulin-independent cyclic nucleotide phosphodiesterase (CaM independent PDE) activity was observed. Perfusion of hearts with cell-permeable calpain inhibitor suppressed the increase of Ca2+ and CaM-independent PDE activity. Protein expression of CaMPDE was uneffected by hypoxic injury to rat myocardium. The purified heart CaMPDE was proteolyzed by calpains into a 45 kDa immunoreactive fragment in vitro. Based on these results, we propose that hypoxic injury to rat myocardium results in the generation of CaM-independent PDE by calpain mediated proteolysis, allowing the maintenance of cAMP concentrations within the physiological range. PMID- 11926173 TI - Universal restriction site-free cloning method using chimeric primers. AB - A universal restriction site-free cloning method has been developed to precisely insert a DNA fragment into a vector at any desired location without altering any nucleotide(s) in either the DNA fragment or the vector. The technique employs two pairs of chimeric primers, each containing a ribonucleotide. One pair of primers is used to amplify a target DNA fragment and another is used to prepare a linear vector. The ribonucleotide is used as a specific site for cleavage promoted by rare-earth metal ions such as La3+ or Lu3+. Therefore, blunt-ended PCR products can be converted into a dsDNA with single-stranded 3'overhangs for efficient ligation. The primers are designed so that both the target DNA fragment and vector PCR products create defined 3' overhangs to permit the formation of a seamless plasmid during the subsequent ligation. This method has been used successfully to clone the E. coli gene coding for peptidyl-tRNA hydrolase. PMID- 11926174 TI - Use of signal-mediated amplification of RNA technology (SMART) to detect marine cyanophage DNA. AB - Here, we describe the application of an isothermal nucleic acid amplification assay, signal-mediated amplification of RNA technology (SMART), to detect DNA extracted from marine cyanophages known to infect unicellular cyanobacteria from the genus Synechococcus. The SMART assay is based on the target-dependent production of multiple copies of an RNA signal, which is measured by an enzyme linked oligosorbent assay. SMART was able to detect both synthetic oligonucleotide targets and genomic cyanophage DNA using probes designed against the portal vertex gene (g20). Specific signals were obtained for each cyanophage strain (S-PM2 and S-BnMI). Nonspecific genomic DNA did not produce false signals or inhibit the detection of a specific target. In addition, we found that extensive purification of target DNA may not be required since signals were obtained from crude cyanophage lysates. This is the first report of the SMART assay being used to discriminate between two similar target sequences. PMID- 11926175 TI - Evaluation of cognitive assessment in stroke rehabilitation. AB - OBJECTIVE: To assess the effect of cognitive assessment on the functional outcome of stroke patients and quality of life for both patients and their carers. DESIGN: A multicentre, single-blind, randomized controlled trial. SETTING AND PARTICIPANTS: Two hundred and twenty-eight stroke patients were recruited from hospital wards in three UK centres. INTERVENTIONS: Patients were screened for cognitive impairment and randomly allocated to either routine care (116 patients) or routine care plus a detailed cognitive assessment (112 patients). MAIN OUTCOME MEASURES: Outcome was assessed three and six months after recruitment by an independent assessor blind to the intervention on Extended ADL, Cognitive Failures Questionnaire, General Health Questionnaire-28 for patients and carers and Carer Strain Index. RESULTS: There was no significant difference between the two groups in patients' functional outcome, perceived cognitive ability, level of psychological distress or satisfaction with care. There was a trend for the assessment group to have lower levels of carer strain (p = 0.06). CONCLUSIONS: The provision of information about cognitive assessment in stroke rehabilitation may decrease carer strain. PMID- 11926176 TI - Measuring nursing needs of stroke patients in clinical rehabilitation: a comparison of validity and sensitivity to change between the Northwick Park Dependency Score and the Barthel Index. AB - OBJECTIVE: To compare the content and convergent validity and the sensitivity to change of the Northwick Park Dependency Score (NPDS) to that of the Barthel Index in patients with stroke. SETTING: Rehabilitation Centre De Hoogstraat, Utrecht, The Netherlands. DESIGN: Inclusion of consecutive patients with stroke. Measurements at admission and every four weeks until final measurement at discharge. Administration by nurses of NPDS, Barthel Index and nurses' global ratings of nursing dependency. RESULTS: Thirty-one patients were included and 123 measurements were obtained. Median Barthel Index scores at admission and discharge were 11.2 and 17.8 respectively. At discharge, 58.6% of all patients obtained the best possible Barthel Index score, against 31.1% for the Basic Care Needs section and 87.1% for the Special Nursing Needs sections of the NPDS. Spearman correlations between NPDS and Barthel Index were very strong (0.82-0.92) at each measurement. Both the NPDS and the Barthel Index showed strong relationships with the global rating of nursing dependency (-0.82 and 0.70 respectively) and showed good sensitivity to change, Z-values for differences between admission and discharge were 4.06 (p < 0.001) for the NPDS and 4.20 (p < 0.001) for the Barthel Index. Most (9/12) nurses involved in this study judged the NPDS to provide better, more precise and complete information about the actual need for care of the patients. CONCLUSIONS: Validity and sensitivity to change of the NPDS were not superior to those of the Barthel Index but most nurses preferred the NPDS for future use. PMID- 11926177 TI - Current practice of prophylactic anticoagulation in Guillain-Barre syndrome. AB - OBJECTIVE: To document the incidence of venous thromboembolism in Guillain-Barre syndrome and current practice of anticoagulating these patients. DESIGN: Retrospective study. SETTING: Acute neurology and rehabilitation wards in a teaching hospital. SUBJECTS: Seventy-three patients with Guillain-Barre syndrome admitted to Addenbrooke's Hospital from 1995 to 1999. RESULTS: Out of 73 patients, 50 were anticoagulated (68%) for 5-490 days with mean of 72 days. Anticoagulation was discontinued in 28 patients when they could walk independently and in six who were still wheelchair dependent. Five patients developed clinical deep venous thrombosis (DVT) (7%), three of them had pulmonary emboli. Venous thrombosis occurred in the first two months after onset in four patients. Two patients were not anticoagulated when they developed their DVT and the other three were on enoxaparin; one of these three had a pulmonary embolism and died. CONCLUSION: Despite prophylactic anticoagulants being used in the majority of patients admitted with major problems of mobility, 6% (3 out of 50) still developed clinically detected DVT and two developed pulmonary embolism. While this incidence is considerably lower than data reported before routine anticoagulation became a standard practice, these data reinforce the need for anticoagulation and suggest that full anticoagulation might be needed to reduce the incidence of thromboembolic complications further. PMID- 11926179 TI - Computer-assisted image analysis of neovascularization in thyroid neoplasms from dogs. AB - OBJECTIVE: To develop a computer-assisted image analysis procedure for quantitation of neovascularization in formalin-fixed paraffin-embedded specimens of thyroid gland tissue from dogs with and without thyroid gland neoplasia. SAMPLE POPULATION: 47 thyroid gland carcinomas, 8 thyroid gland adenomas, and 8 specimens of thyroid tissue from dogs without thyroid gland abnormalities (normal). PROCEDURE: Serial tissue sections were prepared and stained with antibodies against human CD31 or factor VIII-related antigen (factor VIII-rag). The areas of highest vascularity were identified in CD31-stained sections, and corresponding areas were then identified in factor VIII-rag-stained sections. Image analysis was used to calculate the total vascular density in each section, and neovascularization, expressed as a percentage, was determined as the absolute value of the total vascular density derived from factor VIII-rag-stained sections minus the vascular density derived from CD31-stained sections. RESULTS: Mean vascular density of thyroid gland carcinomas derived from CD31-stained sections was significantly greater than density derived from factor VII I-rag-stained sections. This incremental difference was presumed to represent degree of neovascularization. However, significant differences were not detected between vascular densities derived from CD31 and factor VIII-rag-stained sections for either normal thyroid gland tissue or thyroid gland adenomas. No significant correlations were found between vascular density in thyroid gland carcinomas and survival time following surgery. CONCLUSION AND CLINICAL RELEVANCE: A computer assisted image analysis method was developed for quantifying neovascularization in thyroid gland tumors of dogs. This method may allow identification of dogs with tumors that are most likely to respond to treatment with novel antiangiogenesis agents. PMID- 11926178 TI - Bilateral microphthalmos with colobomatous orbital cyst. PMID- 11926180 TI - Evaluation of a bladder tumor antigen test for the diagnosis of lower urinary tract malignancies in dogs. AB - OBJECTIVE: To evaluate the use of a human bladder tumor antigen test for diagnosis of lower urinary tract malignancies in dogs. SAMPLE POPULATION: Urine samples from dogs without urinary tract abnormalities (n = 18) and from dogs with lower urinary tract neoplasia (20) or nonmalignant urinary tract disease (16). PROCEDURE: Test results were compared among groups and among 3 observers. The effects of urine pH and specific gravity, degree of hematuria, and storage temperature and time of urine samples on test results were also assessed. RESULTS: Test sensitivity and specificity were 90 and 94.4%, respectively, for differentiating dogs with lower urinary tract neoplasia from dogs without abnormalities. However, specificity decreased to 35% for differentiating dogs with neoplasia from dogs with nonmalignant urinary tract disease. In dogs with neoplasia, results were significantly affected by degree of hematuria. However, addition of blood to urine from dogs without hematuria had no significant effect on test results. Although intraobserver variation was significant, urine pH, specific gravity, or storage time or temperature had no significant effect on results. CONCLUSIONS AND CLINICAL RELEVANCE: Although this bladder tumor antigen test was sensitive for differentiating dogs with malignancies of the lower urinary tract from dogs without urinary tract disease, it was not specific for differentiating dogs with neoplasia from dogs with other lower urinary tract abnormalities. It cannot, therefore, be recommended as a definitive diagnostic aid for the detection of lower urinary tract malignancies in dogs. PMID- 11926181 TI - Evaluation of rapid staining techniques for cytologic diagnosis of intracranial lesions. AB - OBJECTIVE: To evaluate 4 rapid supravital stains and 3 preparation techniques for use in the intraoperative diagnosis of intracranial lesions. ANIMALS: 10 dogs and 1 cat euthanatized for intracranial lesions. PROCEDURE: Specimens were taken from lesions and slides prepared, using 3 techniques: touch impression, medium pressure impression, or smear preparation. Preparations were then stained with 4 stains: modified Wright stain, May-Grunwald-Giemsa, toluidine blue, and zynostain and examined in a blinded randomized fashion. Cytologic diagnosis was compared with histopathologic diagnosis and classified on the basis of identification of the pathologic process and specific diagnosis into the following categories: complete correlation, partial correlation, or no correlation. RESULTS: An overall diagnostic accuracy of 81% (107/132) was achieved on the basis of a combination of partial and complete correlation. Of the stains examined, modified Wright stain appeared to be most accurate, with complete correlation in 17 of 33 (52%) specimens and partial correlation in 12 of 33 (36%) specimens. Of the preparation methods, touch preparation and smear preparation provided the most accurate results, with an overall diagnostic accuracy of 82% (36/44) for both methods. However, smear preparations appeared to be of greater diagnostic value, with fewer nondiagnostic specimens, compared with touch preparations. CONCLUSIONS AND CLINICAL RELEVANCE: Cytologic preparations provide a useful diagnostic tool for the intraoperative diagnosis of intracranial lesions. All stains examined yielded promising results, the most accurate of which appeared to be the modified Wright stain. The smear preparation appeared to be the preparation method of greatest diagnostic value. PMID- 11926182 TI - Evidence of differences in the biotransformation of organic contaminants in three species of freshwater invertebrates. AB - Acute static bioassays were performed using three freshwater invertebrate species (the oligochaete Lumbriculus variegatus, the fingernail clam Sphaerium corneum and the larvae Chironomus riparius) exposed separately to a variety of 14C radiolabelled contaminants. The aim of this work was to investigate if the chemicals remained as parent compounds after the treatments. Chemicals used were 2,4-dichlorophenol; 2,4,5-trichlorophenol; pentachlorophenol; pyrene; Fenpropidin, and Trifluralin. Homogenates of the whole body tissue of each organism were prepared and total radioactivity was measured. Contaminants were then extracted into organic solvents and analysed by high-pressure liquid chromatography techniques. Chromatograms showed that most of the substances extracted were present as parent compounds in S. corneum and in L. variegatus. In contrast, for C. riparius a low proportion of the chemicals was recovered as parent compounds. These results suggest that different metabolic processes could take place in the different species. PMID- 11926183 TI - Epstein-Barr virus (EBV)-DNA quantification in pediatric allogenic stem cell recipients: prediction of EBV-associated lymphoproliferative disease. PMID- 11926185 TI - Enhanced clearance of leukemic lymphocytes in B-cell chronic lymphocytic leukemia with etodolac. PMID- 11926184 TI - Immunoglobulin (Ig)/BCL6 versus non-Ig/BCL6 gene fusion in diffuse large B-cell lymphoma corresponds to a high- versus low-level expression of BCL6 mRNA. PMID- 11926186 TI - Treatment and outcome of infants with acute myeloid leukemia. PMID- 11926187 TI - Selective serotonin reuptake inhibitors are effective in the treatment of polycythemia vera-associated pruritus. PMID- 11926189 TI - CT-resolution regional hyperthermia treatment planning. AB - Recent development of quasistatic zooming has enabled the computation of mm resolution SAR distributions within reasonable computation times. These high resolution SAR distributions proved to be completely dissimilar from the cm resolution distributions. To study the impact of high-resolution SAR modelling on regional hyperthermia treatment planning (HTP), a conventional, low-resolution treatment plan is compared to an high-resolution plan. This comparison shows that the high-resolution plan yields totally different SAR and temperature distributions when compared to the conventional plan. Both SAR and temperature maxima predicted by the low-resolution plan are not predicted by the high resolution plan and, even worse, the low-resolution plan fails to predict maxima that are predicted by the high-resolution plan. Furthermore, it appears that small-scale SAR maxima can result in temperature maxima which may cause treatment limiting hot spots. These small-scale SAR maxima appear to be highly determined by the dielectric geometry of the patient. This demonstrates the need for an accurate, high-resolution description of this dielectric geometry. Moreover, it suggests that it may be very difficult to reduce potential treatment-limiting hot spots in clinical practice. This study demonstrates the need for high-resolution regional hyperthermia treatment planning. PMID- 11926188 TI - The Fanconi anemia cell line HSC536N is not sensitive to interferon-gamma and does not cleave PARP in response to FAS-mediated cell killing. PMID- 11926190 TI - A flexible optimization tool for hyperthermia treatments with RF phased array systems. AB - In hyperthermia treatments performed with a radio-frequency phased array, the main issue to apply the excitation amplitudes and phases of the applicators for which tumour heating is optimal, i.e. the maximal therapeutic gain without unwanted side effects. Due to the complex interaction of the radiated EM-field and the patient's tissues, it is very difficult to find these optimal excitation (amplitude and phase) parameters by intuition. Calculation of the EM-field distribution within the patient can aid in finding the optimal excitation setting. However, this remains a difficult task because of the degrees of freedom available (2n - 1, with n the number of applicators in the array) and because a large temperature elevation may occur at healthy tissue sites resulting in unwanted side effects, e.g. pain or healthy tissue damage. Therefore, determining the excitation amplitudes and phases yielding optimal tumour heating can be done effectively only by application of a computerized optimization procedure. Optimization of the temperature distribution in the patient requires detailed knowledge of the thermal tissue parameters. Techniques for determining these properties are not commonly available and the use of averaged values for parameters like the tissue perfusion is expected to introduce large errors for individual patient treatment planning. As a consequence, the SAR distribution, being proportional to the temperature increase at treatment start, is more often selected for optimization. The 'optimized' excitation amplitudes and phases are found by maximization of a certain SAR ratio. Several propositions for this SAR ratio have been reported in the literature, e.g. the ratio of the SAR at the tumour site and the SAR at sites where unwanted side effects may occur. However, the definition of these ratios does not constrain the SAR value at these tissue locations to a safe value. In this paper, a tool for the optimization of the SAR distribution including the specification of constraints is presented. The tool focuses on the definition of the average SAR as a function of the excitation amplitudes and phases in a volume of arbitrary size (e.g. the tumour volume or the whole patient volume). These functions can be applied in either customized or commercially available optimization routines and they enable the definition of constraints for the average SAR in a certain volume. The described tool is illustrated for a patient case, showing the flexibility and easy application of the tool. PMID- 11926191 TI - Self-care in adults with sickle cell disease. AB - This article summarizes the psychometric evaluation of the Chronic Illness Assessment Interview for Sickle Cell Disease (CIAI-SCD), an instrument based on a model of self-care for adult patients with chronic medical conditions. The CIAI SCD was administered to 104 adults with sickle cell disease. A factor analysis identified three factors that reflected the psychological constructs of Personal Satisfaction and Perceived Control (Factor 1), Feeling Concerned and Worried (Factor 2), and Feeling Supported (Factor 3). Preliminary evidence for the internal consistency, test-retest reliability content validity, and construct validity of the CIAI-SCD was obtained. After further refinement and validation, the CIAI-SCD may be a useful tool for assessing factors related to self-care skills among adults with sickle cell disease. PMID- 11926192 TI - Effects of carbon nutrition on the physiological expression of HCO3- transport and the CO2-concentrating mechanism in the Cyanobacterium chlorogloeopsis sp. ATCC 27193. AB - We have examined the effect of inorganic and organic carbon nutrition on the physiological expression of HCO3- transport and the CO2-concentrating mechanism (CCM) in the nutritionally versatile cyanobacterium Chlorogloeopsis sp. ATCC 27193. Cells grown under photoautotrophic conditions in the presence of limiting or replete levels of inorganic carbon (Ci), or grown under mixotrophic (light) or chemoheterotrophic (dark) conditions in the presence of sucrose retained both active CO2 and Na(+)-independent HCO3- transport activity. However, two distinct effects on the kinetic properties of HCO3- transport were observed, which segregated on the basis of phototrophic and chemoheterotrophic growth in the dark. In the former, the apparent substrate affinity of the HCO3- transport system (K0.5) varied (12-fold) in response to the growth Ci or mixotrophy while the maximum rate of HCO3- transport was approximately constant. In the latter case, the K0.5 value was unchanged from the starting value (35 microM) of Ci limited photoautotrophic cells used to initiate the dark-grown cultures, but transport capacity declined 3-fold. Modulation of the K0.5 (HCO3- transport) value required light. Cellular carboxysome content was unaffected by growth under any of the regimes employed and these structures were the predominant location of ribulose-1,5-bisphosphate carboxylase/oxygenase, as indicated by immunogold electron microscopy. Mixotrophic and chemoheterotrophic growth resulted in a diminished ability to concentrate Ci internally and a reduction in Ci accumulation ratios at low external Ci concentrations. The relationship between photosynthetic carbon fixation and the internal Ci pool varied by 2-fold, with high-Ci-grown cells being the most efficient and mixotrophically grown cells the least, indicating that there was limited capacity to modulate this relationship in response to changes in carbon nutrition. Within broad limits this relationship appeared to be a fixed trait of the strain and an important factor in determining growth rate. PMID- 11926193 TI - Common complications of body piercing. PMID- 11926194 TI - Care of older people. Staff could be using wrong DoH guidance. PMID- 11926195 TI - Spatial characteristics of fine particulate matter: identifying representative monitoring locations in Seattle, Washington. AB - This study investigates how PM2.5 varies spatially and how these spatial characteristics can be used to identify potential monitoring sites that are most representative of the overall ambient exposures to PM2.5 among susceptible populations in the Seattle, WA, area. Data collected at outdoor sites at the homes of participants of a large exposure assessment study were used in this study. Harvard impactors (HIs) were used at 40 outdoor sites throughout the Seattle metropolitan area. Up to six sites at a time were monitored for 10 consecutive 24-hr average periods. A fixed-effect analysis of variance (ANOVA) model that included date and location effects was used to analyze the spatial variability of outdoor PM2.5 concentrations. Both date and location effects were shown to be highly significant, explaining 92% of the variability in outdoor PM2.5 measurements. The day-to-day variability was 10 times higher than the spatial variability between sites. The site mean square was more than twice the error mean square, showing that differences between sites, while modest, are potentially an important contribution to measurement error. Variances of the model residuals and site effects were examined against spatial characteristics of the monitoring sites. The spatial characteristics included elevation, distance from arterials, and distance from major PM2.5 point sources. Results showed that the most representative PM2.5 sites were located at elevations of 80-120 m above sea level, and at distances of 100-300 m from the nearest arterial road. Location relative to industrial PM2.5 sources is not a significant predictor of residential outdoor PM2.5 measurements. Additionally, for sites to be representative of the average population exposures to PM2.5 among those highly susceptible to the health effects of PM2.5, areas of high elderly population density were considered. These representative spatial characteristics were used as multiple, overlapping criteria in a Geographic Information System (GIS) analysis to determine where the most representative sites are located. PMID- 11926196 TI - Staffing shortages. Charting workforce woes. PMID- 11926197 TI - Apodized fiber Bragg gratings manufactured with the phase plate process. AB - We present a manufacturing method based on the dynamic use of phase plates to photowrite Bragg gratings. This process allows for control of the local value of the index modulation envelope in the grating. The application to apodized fiber Bragg gratings is discussed. PMID- 11926198 TI - Sex pheromone of South American tortricid moth Argyrotaenia sphaleropa. AB - By means of electroantennographic detection and gas chromatography-mass spectrometry, the sex pheromone of Argyrotaenia sphaleropa was identified as a mixture of (Z)-11-tetradecenal, (Z)-11,13-tetradecadienal, (Z)-11-tetradecenyl acetate, and (Z)-11,13-tetradecadienyl acetate in the ratio of 1:4:10:40. Best trap catches were obtained with mixtures of (Z)-11-tetradecenal and (Z)-11,13 tetradecadienal in the ratio of 1:4 to 1:9. PMID- 11926199 TI - Conformational analysis: a new approach by means of chemometrics. AB - In conformational analysis, the systematic search method completely maps the space but suffers from the combinatorial explosion problem because the number of conformations increases exponentially with the number of free rotation angles. This study introduces a new methodology of conformational analysis that controls the combinatorial explosion. It is based on a dimensional reduction of the system through the use of principal component analysis. The results are exactly the same as those obtained for the complete search but, in this case, the number of conformations increases only quadratically with the number of free rotation angles. The method is applied to a series of three drugs: omeprazole, pantoprazole, lansoprazole-benzimidazoles that suppress gastric-acid secretion by means of H+, K+-ATPase enzyme inhibition. PMID- 11926200 TI - Longitudinal relationship between pain and depression in older adults: sex, age and physical disability. AB - OBJECTIVE: Better understanding of the relationship between pain and depression in older adults in the community is of particular importance considering the high prevalence of both conditions in these adults. In the present study, the longitudinal relationship between pain and depression in older adults was examined, thereby taking into account the role of physical disability and the possibly modifying effect of sex and age. METHODS: The study is based on a sample which at the outset consisted of 325 non-depressed and 327 depressed persons (55 85) drawn from a larger random community-based sample in the Netherlands. Depression (CES-D) and pain (subscale of the Nottingham Health Profile) were measured at eight successive waves over 3 years. RESULTS: Pain was very persistent over time as was to a lesser extent depression. The prognosis of comorbid pain and depression was poor. In longitudinal analyses (Generalized Estimating Equations), pain and depression were strongly associated. At the symptom level, the pain-depression relationship was found to be stronger in men than in women. There was no effect of age on the pain-depression relationship. No support was found for the hypothesis that the pain-depression relationship is mediated by disability. CONCLUSION: The persistent nature of pain and to a lesser extent depression and the intimate and probably reciprocal association between them stress the need for adequate treatment strategies. PMID- 11926201 TI - Bone involvement in idiopathic hypercalciuria. AB - BACKGROUND: To evaluate bone involvement in idiopathic hypercalciuria, 40 lithiasic patients and 10 controls were studied. METHODS: According to urinary calcium excretion, patients were first classified as hypercalciuric (Hca, n = 22) and normocalciuric (Nca, n = 18). The Hca patients were then subclassified according to bone densitometry (BMD) as osteopenic (HcaO, n = 10) and non osteopenic (HCaNO, n = 12). Routine biochemistry, dietary records, bone histomorphometry. and cytokines (IL-1beta, IL-6, and TNF) production by peripheral blood mononuclear cell cultures were studied. RESULTS: There were no differences in routine biochemistry between Hca and Nca groups, except for urinary calcium. Inadequate nutrition was observed in Hca group, showing high protein (80.9% of the patients), carbohydrate (76.2%) and sodium (90%) intake. Calcium intake was low in Hca (57%) and Nca (83%) groups. IL-6 and TNF were not different between the Hca and Nca groups. IL-1beta levels were significantly high in both groups when compared to controls. IL-6 and TNF were higher in HcaO than Nca. BMD in femoral neck in HcaO was lower than in HcaNO and Nca groups. Eroded surface (ES/BS) increased in 91% of the Hca group and 36% had a mineralization defect. In the HcaO group serum PTH correlated negatively with trabecular bone volume (BV/TV) and positively with ES/BS. 1,25(OH),D3 levels correlated positively with osteoblastic surface. Calcium intake correlated positively with BV/TV and inversely with ES/BS. A negative correlation was observed between IL-6 levels and Z score of the femoral neck. CONCLUSION: Bone involvement was detected in a young population with nephrolithiasis demonstrating that a strict follow-up is necessary in order to control hypercalciuria. PMID- 11926202 TI - Hypertension after renal transplantation and polymorphism of genes involved in essential hypertension: ACE, AGT, AT1 R and ecNOS. AB - BACKGROUND: Arterial hypertension (HT), secondary to cyclosporine A (CsA) used as main immunosuppressive treatment in renal transplantation (RTx), is very frequent (70%), usually severe and explained mostly by vasoconstriction of the glomerular afferent arteriole with secondary sodium and water retention. MATERIAL AND METHODS: In a retrospective study, we have analyzed 294 consecutive recipients receiving a first renal cadaveric allograft and all treated with CsA (the majority with triple therapy). We studied, by molecular biology, the polymorphism of genes previously implicated in essential HT such as: angiotensin-converting enzyme (ACE: II, ID and DD), angiotensinogen (AGT: MM, MT and TT), angiotensin II type 1 receptor (AT1-R: AA, AC and CC) and endothelial constitutive nitric oxide synthase (ecNOS: aa, ab and bb), and correlated the data to the prevalence and severity of post-Tx HT. This cohort included 195 (66%) males and 99 females with a mean age of 42 years at time of Tx. The presence and severity of post-Tx HT were indicated by initial persistent blood pressure over 140/90 mmHg with the need for at least one anti-hypertensive drug and by the number of anti-HT medications required to achieve its control. RESULTS: The distribution of the specific alleles and genotypes for ACE, AGT, AT1-R, and ecNOS was not different in transplant recipients compared to 181 controls. At 5 years post-Tx, the prevalence of HT was 72% (169 out of 235) among functioning grafts. There was no significant difference for ACE, AGT, AT1R and ecNOS genotypes distribution between hypertensive vs non-HT recipients. The number of anti-hypertensive drugs prescribed was not different among ACE, AGT, and AT1-R genotypes. However, the a allele and the non-bb genotype (aa + ab) for ecNOS were significantly (p = 0.001) associated with a less severe HT, needing fewer anti-HT drugs. At 10 years post Tx, the HT prevalence remained high 78% (67 out of 86) among functioning Tx. However, the limited numbers did not allow further correlation. CONCLUSIONS: This study produced mainly negative results except for ecNOS-a allele, which seems to protect against severe hypertension. The explanation remains speculative but probably relates to the known cyclosporine-induced upregulation of ecNOS gene and enzyme activity. PMID- 11926203 TI - [Effect of oral administration of ascorbic acid on insulin sensitivity and lipid profile in obese individuals]. AB - BACKGROUND: The aim of this study was to identify the effect of an oral ascorbic acid (AA) supplement on lipid profile and insulin sensitivity in obese people. RESEARCH DESIGN AND METHODS: A randomized double-blind clinical trial placebo controlled was performed in 16 obese male volunteers [body mass index (BMI) 30-40 kg/m2]. Eight received orally 1 g of AA daily for four weeks and the other eight volunteers received placebo by the same scheme and period of time. Before and after the pharmacological intervention were measured total cholesterol, high density-lipoprotein (HDL) cholesterol, triglycerides, glucose, creatinine and uric acid. Low-density-lipoprotein (LDL) cholesterol and very-low-density lipoprotein (VLDL) triglycerides were calculated using formulas. In order to assess insulin sensitivity before and after the intervention, the steady-state glucose (SSG) was calculated from the insulin suppression test modified with octreotide. RESULTS: There were not significant differences in clinical characteristics between both groups. Basal metabolic profile and SSG were similar between both groups. There were not significant differences in both groups between before and after the intervention in metabolic profile and insulin sensitivity. CONCLUSION: AA did not modify the lipid profile nor insulin sensitivity in the group of obese people studied. PMID- 11926204 TI - Salvia lavandulaefolia essential oil inhibits cholinesterase in vivo. AB - The essential oil of Salvia lavandulaefolia at two dosage levels was administered orally to rats for five days. Choline esterase activity was measured post mortem in three areas of the brain, both in the absence and presence of TEPP, a specific butylcholine esterase inhibitor, and was found to be significantly reduced in the striatum with both doses and also in the hippocampus at the higher dose. The activity of the enzyme in the cortex was not significantly reduced even at the higher dose. Thus it appears that S. lavandulaefolia oil, shown to inhibit choline esterase in vitro, also has an in vivo effect and this may help explain its traditional use for ailing memory. PMID- 11926205 TI - Mutational analysis of the GNAS1 exons encoding the stimulatory G protein in five patients with pseudohypoparathyroidism type 1a. AB - We analyzed the GNAS1 gene in five patients with pseudohypoparathyroidism type 1a (PHP1a) by performing polymerase chain reaction, followed by sequencing all 13 exons of the gene, single-stranded conformational polymorphism (SSCP) or heteroduplex analysis (HD). Three novel mutations were discovered: (1) a de novo 3 bp insertion of CTG in codon 47 of exon 1; (2) a missense mutation 1103T in exon 4; and (3) a de novo mutation of Arg280Gly in exon 10. Two other mutations, previously described in the literature, include: (1) a de novo 4 bp deletion (deltaGACT) involving codons 189 and 190 in exon 7, and (2) a deletion of a cytosine nucleotide at codon 115 in exon 5. We conclude that mutational analysis of the GNAS1 gene is a strong supportive tool for the diagnosis of PHP1a, and is a useful adjunct to the synthetic parathyroid hormone infusion test for PTH resistance. PMID- 11926206 TI - Hypopituitarism, deficiency of factors V and VIII and von Willebrand factor: an uncommon association. AB - A 9 year-old boy with hypopituitarism and blood coagulation abnormalities is presented and discussed. The association between acquired von Willebrand disease and hypothyroidism has been reported but the combination of hypopituitarism and coagulopathy is unusual. Combined multiple clotting deficiencies are rare and, when present, factors V and VIII is the commonest association. Although it is known that hypothyroid patients may have a decrease in von Willebrand's factor (vWf) and factor VIII, there are no reports of hypopituitarism associated with combined deficiency of factors V, VIII, and vWf. PMID- 11926207 TI - CNS germinoma in a boy with simple virilizing 21-hydroxylase deficiency and precocious puberty. AB - A 6 year-old boy presented with peripheral precocious puberty and was diagnosed as having simple virilizing 21-hydroxylase deficiency based on clinical features and elevated 17-hydroxyprogesterone levels on ACTH stimulation. He was managed with glucocorticoids and mineralocorticoids. Two years later he presented with features of CNS involvement in the form of seizures and raised intracranial pressure with rapid progression of puberty. Contrast enhanced CT scan of brain showed an intraventricular tumor with cerebrospinal fluid cytology suggestive of germinoma. Serum and CSF levels of human chorionic gonadotropin (hCG) and alphafetoprotein (AFP) were elevated, confirming the diagnosis of germinoma. PMID- 11926208 TI - Outcomes of infants born to mothers with inflammatory bowel disease: a population based cohort study. AB - OBJECTIVE: Limited population-based data on inflammatory bowel disease (IBD) and pregnancy outcomes exist. The purpose of this study is to determine the association between maternal IBD status and adverse pregnancy outcomes. METHODS: Using computerized birth records of infants born to mothers with Crohn's disease (CD) or ulcerative colitis (UC) and mothers without diagnoses of IBD (no-IBD) in Washington State, we performed a cross-sectional retrospective study to determine gestational age, birth weight, and congenital malformations. RESULTS: Preterm delivery was seen in 15.2% of CD births, 10.4% of UC births, and 7.2% of no-IBD births. Low birth weight was found in 16.8% of CD births, 7.6% of UC births, and 5.3% of no-IBD births. Smallness for gestational age was present in 15.2% of CD births, 10.5% of UC births, and 6.9% of no-IBD births. Only CD births were at significantly increased risk of preterm delivery (p < 0.0025), low birth weight (p < 0.001), and smallness for gestational age (p < 0.001). Congenital malformations were more commonly recorded in UC births than in controls (7.9% vs 1.7%, p < 0.001), whereas 3.4% of CD births had malformations recorded. Using multivariable logistic regression, CD births were more likely to be preterm (odds ratio [OR] = 2.3, 95% CI = 1.4-3.8) and have low birth weights (OR = 3.6, CI = 2.2-5.9) and smallness for gestational age (OR = 2.3, CI = 1.3-3.9). UC births were more likely to have congenital malformations reported (OR = 3.8, CI = 1.5 9.8). CONCLUSIONS: Maternal IBD is associated with increased odds of preterm delivery, low birth weight, smallness for gestational age (CD), and reporting of congenital malformations (UC). PMID- 11926209 TI - Central processing of rectal pain in patients with irritable bowel syndrome: an fMRI study. AB - OBJECTIVES: In healthy subjects, the neural correlates of visceral pain bear much similarity with the correlates of somatic pain. In patients with irritable bowel syndrome, the central nervous system is believed to play a strong modulatory or etiological role in the pathophysiology of the disease. We hypothesize that this role must be reflected in aberrations of central functional responses to noxious visceral stimulation in these patients. To verify this hypothesis, we have induced transient rectal pain in patients and assessed the functional responses of the brain by means of functional magnetic resonance imaging. METHODS: Twelve right-handed patients (11 female) were examined. Functional imaging (1.5 T) was performed following a block paradigm, alternating epochs with and without noxious stimulation of the rectum. Rectal pain was induced by inflating a latex balloon. Whole-brain coverage was achieved by means of echo-planar magnetic resonance acquisition. RESULTS: A strong variability of the individual responses to rectal pain was found in patients with irritable bowel syndrome. Significant activations were found in only two patients, and group analysis did not reveal significant activations. In contrast, all patients exhibited significant deactivations. Group analysis revealed significant deactivations within the right insula, the right amygdala, and the right striatum. CONCLUSIONS: This study reveals aberrant functional responses to noxious rectal stimulation in patients with irritable bowel syndrome. Those results add grounds to the hypothesis that the central nervous system plays a significant role in the pathophysiology of this syndrome. PMID- 11926210 TI - High prevalence of undetected ulcerative colitis: data from the Nottingham fecal occult blood screening trial. AB - OBJECTIVES: Inflammatory bowel disease (IBD) is usually diagnosed as a result of symptoms but occasionally is found during investigation for other conditions. An earlier report from Nottingham had found a high prevalence of previously undetected "asymptomatic" IBD detected as a result of colorectal cancer screening, and the aim of this study was to reassess the prevalence, symptoms, and outcome in these patients. METHODS: We investigated subjects found to be fecal occult blood (FOB) positive in a randomized trial of FOB screening for colorectal cancer. All FOB-positive subjects were investigated by colonoscopy or flexible sigmoidoscopy and barium enema. Subjects with IBD were referred back to their general practitioner for any further investigation and treatment. RESULTS: Seventy-five thousand two hundred fifty-three subjects (aged 45-74) were sent FOB tests and 44,838 (60%) completed a series of tests on one or more occasions. Of 133,000 test series, 1.5% were positive. During investigation 53 cases of previously undetected IBD (52 of ulcerative colitis) were found; 52% (27/52) had proctosigmoiditis only, whereas 25% (13/52) had pancolitis. Only 17% (9/52) were completely asymptomatic, with a half or more reporting some rectal bleeding (54%) or diarrhea (50%). The overall prevalence of undetected ulcerative colitis was 69/10(5) (95% CI = 50-88/10(5)) in people offered screening and 116/10(5) (95% CI = 85-147/10(5)) in people accepting screening and was higher in men. Of 32 subjects followed up 2-12 yr after diagnosis, 91% (29) continued to have few or no symptoms, with only 12 currently receiving any treatment for their colitis. CONCLUSIONS: In comparison with detected disease, undetected ulcerative colitis is relatively common but does usually cause some symptoms. It generally appears to follow a benign course, but a significant proportion have extensive colitis and may therefore be at an increased risk of colorectal cancer. PMID- 11926211 TI - Metastatic breast carcinoma presenting as acute liver failure and portal hypertension. PMID- 11926212 TI - Lamivudine transiently reduces viral load and improves liver function in liver transplant recipients with fibrosing cholestatic hepatitis. PMID- 11926213 TI - Health care experiences and preferences of uninsured workers. AB - An exploratory study of the experiences, beliefs, and preferences of uninsured workers found that uninsured workers are active in seeking solutions to their health care needs and have decided opinions about health care. They use a cost benefit analysis in deciding which health problems warrant professional attention. However, uninsured workers also exhibit a contradictory pattern of behaviors and beliefs. They say their health is good, yet many have illnesses that they neglect. They are satisfied with the health care services they use, yet they want health insurance because they believe it provides more choice and ensures better care and treatment. They express few negative feelings about Medicaid, yet even among the poorest of these workers, few have recently used Medicaid. These findings can inform policy options being considered. PMID- 11926215 TI - Placing social work in time and space. PMID- 11926214 TI - Managed care and the judicial system: another avenue for reform? AB - Over the past several years the judiciary has become a major venue for challenging managed care. This article examines the present and potential effect of such litigation, focusing on the recent Supreme Court decision in Pegram v. Herdrich and class action litigation-filed under the Racketeer Influenced and Corrupt Organizations Act. The author concludes that although court-based reforms are a necessary and pragmatic response to managed care, they can result in only incremental changes. Litigation also cannot resolve the larger conflicts that result when a social welfare service such as health care is provided through a corporate model. Social workers need to look beyond the "next best" reform of managed care and advocate for more fundamental solutions to the inequities and inefficiencies in the provision of health care. PMID- 11926217 TI - Group therapy intervention for male batterers: a microethnographic study. AB - Treatment of male batterers has been a controversial issue for study and practice in the field of behavioral health care. Empirical evidence on success of treatment of male abusers is mixed. Little is known about what makes a treatment successful for an individual abusive man. Knowledge of how batterers learn and change in therapy and how they apply this learning in their day-to-day lives can be useful in designing successful treatments. The microethnographic study of a group therapy for male batterers discussed in this article describes change processes in abusers and examines the dynamics of unsuccessful processes. In doing so, this study provides a useful evaluation of group therapy as it is offered to batterers. The authors conclude by discussing the implications of microethnographic methods for social work students learning to evaluate their practice. PMID- 11926216 TI - The role of cognitive control in mediating the effect of stressful circumstances among Korean immigrants. AB - The study reported in this article investigated relationships among stressful circumstances, cognitive control (the individual's perception of control over life), and distress among Korean immigrants in the United States. Specifically, it was hypothesized that cognitive control mediated the effect of exposure to stressful circumstances on distress. A total of 159 Korean immigrants participated in the study. The result of the study provided strong support for the role of cognitive control in mediating the relationship between stressful circumstances and distress. The detrimental effect of exposure to stressful circumstances on distress was weakened by cognitive control. Implications for social work practice and future research are discussed. PMID- 11926218 TI - The crisis of public health revisited: implications for social work. PMID- 11926219 TI - HIV-prevention capacity building in gay, racial, and ethnic minority communities in small cities and towns. PMID- 11926220 TI - Evaluating HIV mental health training: changes in practice and knowledge for social workers and case managers. PMID- 11926221 TI - The impact of hospital restructuring on social work field education. AB - Hospital restructuring has had a dramatic impact on social work practice and field education. In a qualitative focus group study of nine educational coordinators responsible for educational programs in teaching hospitals with 25 to 100 social workers each, four main themes emerged: (1) difficulty managing field education when the actual change experienced in restructuring hospitals was far less progressive and systemic than the hospital missions espoused; (2) a struggle to maintain stable student programs when change was unpredictable; (3) the importance of support, reciprocity, and advocacy from the university; and (4) the need to be creative in delivering the educational program while undergoing organizational change. PMID- 11926222 TI - Tech knowledge: introducing computers for coordinated care. PMID- 11926223 TI - Using customer satisfaction surveys to improve quality of care in nursing homes. PMID- 11926224 TI - Induction in variants of White's effect: common or separate mechanisms? AB - In White's display the gray target surrounded more by black than white appears darker than the target of the same physical luminance surrounded more by white than black. Several subsequent studies have shown that this effect occurs only when the luminance of the test regions lies between the minimum and maximum luminance values of the inducing stripes. With targets either lighter or darker than both inducing stripes, the direction of the effect is reversed and the effect is known as the 'inverted' White's effect. Views differ on whether the classical and inverted White's effects are mediated by common or separate underlying mechanisms. We varied the aspect ratio of the test and inducing regions in the classical and inverted White's effects. Consistent with previously reported findings, we found that the direction of the classical effect did not depend on the amount of black or white border in immediate contact with the test patch. On the other hand, perceived lightness in the inverted White's effect was affected by such variations, suggesting that induction in classical and inverted White's configurations is governed by different mechanisms. These results confirm the critical importance of the interaction between luminance and geometric relationships in induced brightness. PMID- 11926225 TI - Painting with light by Rob and Nick Carter: dramatic failures of colour constancy in articulated scenes. AB - Recent work by British artists Rob and Nick Carter uses kinetic lights illuminating abstract photographic prints to induce dramatic failures of colour constancy. PMID- 11926227 TI - Hospital chaplain. PMID- 11926226 TI - James Herrick and the description of sickle-cell disease. PMID- 11926228 TI - Essential research issues in rural health: the state rural health directors' perspective. PMID- 11926229 TI - How state rural health directors obtain policy-relevant research information. PMID- 11926230 TI - Averting malpractice by information: informed consent in the pediatric treatment environment. PMID- 11926231 TI - Hall v. Anwar. PMID- 11926232 TI - Opportunity: creating a national oral health plan. PMID- 11926233 TI - The Vipeholm Dental Caries Study: recollections and reflections 50 years later. PMID- 11926235 TI - Mercury uptake and kinetics after ingestion of dental amalgam. AB - The aim of the present study was to investigate the G-1 uptake of mercury (Hg) after intake of a single dose of amalgam-Hg, followed by pharmacokinetic analysis of the data. Eleven volunteers without amalgam fillings ingested 1.00 g amalgam powder. Hg in plasma vs. time was analyzed with a two-compartment model by means of mixed-effects modeling. A fraction of the absorption rate of Hg to the central compartment was inversely proportional to the plasma ferritin levels. The population mean half-life of the terminal phase of Hg in plasma was 37 days, with a considerable standard deviation in the population. The absorbed fraction of the administered dose was estimated to be about 0.04%. It is concluded that the G-1 uptake of Hg is of quantitative importance during dental treatment. PMID- 11926234 TI - Effects of dental resins on TNF-alpha-induced ICAM-1 expression in endothelial cells. AB - Many reports have demonstrated inflammation after the placement of dental restorations. To explain this side-effect, we studied a biomarker in the inflammatory response. The intercellular adhesion molecule-1 (ICAM-1) is a key mediator for recruitment of leukocytes to the site of inflammation. Therefore, we investigated whether methacrylates (a BISGMA-based dental resin, BISGMA, and MAA) and Cyracure UVR 6105, an epoxy monomer, could alter ICAM-1 expression in unstimulated and TNF-alpha-stimulated endothelial cells. Six-well plates with monolayers of human umbilical vein cells, ECV 304 (ATCC CRL 1998), were exposed to TNF-alpha (1 ng/mL) in the presence and absence of subtoxic and TC50 doses of chemicals for 24 hrs at 37 degrees C/5% CO2. Several doses of TNF-alpha (0.5-2 ng/mL) were coincubated with 100 microL of undiluted aqueous dental resin extracts. Cells were harvested and stained with mAB FITC-conjugated anti-human ICAM-1 (CD54). ICAM-1 expression was measured by flow cytometry. Cells expressed basal levels of ICAM-1, which was up-regulated by TNF-alpha but was not changed by all samples studied. Except for UVR 6105, the methacrylates significantly decreased ICAM-1 expression in TNF-alpha-stimulated cells. These findings suggest that methacrylates may decrease the recruitment of leukocytes to sites of inflammation. PMID- 11926236 TI - Laser-matrix-fluoride effects on enamel demineralization. AB - Laser and fluoride have been shown to inhibit enamel demineralization. However, the role of organic matrix and their interactions remains unclear. This study investigated the interaction among CO2 laser irradiation, fluoride, and the organic matrix on the demineralization of human enamel. Twenty-four molars were selected and cut into halves. One half of each tooth was depleted of its lipid and protein. The other half served as a matched control. Each tooth half had two window areas, treated with a 2.0% NaF gel. All left windows then received a laser treatment. Next, the tooth halves were subjected to a four-day pH-cycling procedure that created caries-like lesions. Tooth sections were cut from the windows, and microradiographs were used for quantification of the demineralization. The combined fluoride-laser treatment led to 98.3% and 95.1% reductions in mineral loss for enamel with and without organic matrix, respectively, when compared with sound enamel. PMID- 11926237 TI - Molecular structure of acid-etched dentin smear layers--in situ study. AB - It is commonly reported that acid etchants remove the smear layer, but to date, there has been no chemical evidence to support these observations. The purpose of this study was to determine the molecular structure of acid-etched carbide- and diamond-bur-created smear layers. This project tested the null hypothesis that such smear layers are totally removed with current etchants. Smear layer/demineralized/mineralized dentin interfaces were analyzed at 1- m intervals by micro-Raman spectroscopy. Features associated with the organic component were substantially broadened with loss of fine structure, and mineral peaks were clearly evident in the spectra of acid-etched smear layers. The organic features in the spectra of the EDTA-treated smear layer showed relative intensity ratios similar to demineralized dentin without contribution from the mineral phase. The disorganized collagen within the smear layer was not removed but was denatured by the acid treatment; the mineral was trapped in this gelatinous matrix and shielded from complete reaction. PMID- 11926239 TI - Effects of heat treatments on mechanical strength of laser-welded equi-atomic AuCu-6at%Ga alloy. AB - There is little information available on the mechanical strengthening of laser welded gold alloys to achieve reliable dental prostheses. This study examined the hypothesis that heat treatments increase the mechanical strength of a laser welded equi-atomic AuCu-6at%Ga alloy with age "hardenability" at intra-oral temperature. Cut cast gold alloy plates were laser-welded. The specimens were given one of three heat treatments: (1) solution treatment, (2) high-temperature aging after solution treatment, and (3) simulated intra-oral aging after solution treatment. As-cast and uncut specimens were also prepared. Tensile testing was conducted, and the breaking stress and yield strength were recorded. The yield strength values of all the heat-treated specimens nearly reached the values of the corresponding heat-treated control specimens. The results of this study indicated that, for high mechanical strength to be achieved, the laser-welded alloy tested should be aged at a high temperature or be intraorally aged after being laser-welded. PMID- 11926238 TI - The glass-ionomer phase in resin-based restorative materials. AB - Glass-ionomer (GI) fillers are added to restorative materials, but it is unclear if they truly react with these materials. This TEM study evaluated the existence of the GI phase in a conventional GIC (ChemFlex), a resin-modified GIC (Fuji II LC), a giomer (Reactmer Paste), a compomer (Dyract AP), and a composite (SpectrumTPH), before and after water uptake. Wafers were stored at 100% RH for 24 hrs, or in water for 7 or 84 days. ChemFlex glass particles were surrounded by 300-nm-thick silica gel layers. In Fuji II LC, we found thinner hydrogel layers (100 nm) that became thicker upon water storage. No appreciable change occurred in Reactmer Paste. Only a very thin hydrogel layer occurred in Dyract AP, and none was seen in SpectrumTPH after water storage for 84 days. We conclude that the variable extent of the GI phase is determined by differences in the resin composition of the restoratives. PMID- 11926240 TI - Low metacarpal bone density, tooth loss, and periodontal disease in Japanese women. AB - The relationship between periodontitis and systemic bone mineral density in Japanese women is undetermined. We tested the hypothesis that periodontitis was more frequent in women with low metacarpal bone mineral density (m-BMD). Subjects were 190 Japanese women (89 premenopausal, 101 post-menopausal). Periodontal status was evaluated according to the Community Periodontal Index of Treatment Need (CPITN). M-BMD was measured by computed x-ray densitometry. The proportion of subjects with periodontitis (CPITN > or = 3) increased as m-BMD decreased in pre-menopausal (18.2%, 36.9%, and 66.6% in the normal, borderline, and very low m BMD groups, p < 0.02) and post-menopausal women (41.5%, 54.8%, 60%, and 68.4% in the normal, borderline, low, and very low m-BMD groups, p < 0.05). Among post menopausal women, those with very low m-BMD had fewer teeth present than women with normal m-BMD (19.9+/-7.2 vs. 25.1+/-4.1, p < 0.01). These results indicate that m-BMD loss is associated with periodontitis in Japanese women, and with tooth loss after menopause. PMID- 11926241 TI - The trend and risk factors of perceived toothache among Finnish adolescents from 1977 to 1997. AB - Toothache is a subjective oral health indicator that should become uncommon when oral health is improving. The aim of this study was to assess changes in perceived toothache between 1977 and 1997 among Finnish adolescents. In the Adolescent Health and Lifestyle Survey, a self-administered questionnaire was mailed to a representative sample of 12-, 14-, 16-, and 18-year-old Finns every second year. The sample sizes in the surveys varied from 2422 to 11,105, making a total of 35,349 subjects in the entire study. The incidence of toothache during the previous 2 years was requested in 1977, 1985, 1991, 1995, and 1997, and the prevalences were 31%, 25%, 28%, 29%, and 37%, respectively. The increase between 1995 and 1997 was highest among 16- and 18-year-olds. There was no tendency for perceived toothache to decline over the study period, despite the decrease in caries experience. Toothache varied by age, socioeconomic status (SES), place of residence, and toothbrushing frequency. The increase in the incidence of toothache reported after 1995 could be a warning signal that economic recession in Finland has caused changes in the dental care system. PMID- 11926242 TI - The predominant aciduric microflora of root-caries lesions. AB - The etiology of root caries is not fully understood, and although mutans streptococci, lactobacilli, and A. naeslundii have been implicated in its initiation and progression, this study was designed to determine the potential role of other microbial species and the nature of predominant aciduric microflora in the root caries process. We isolated the predominant aciduric microflora from root-caries lesions (n = 14) and sound root surfaces in subjects with (n = 13) or without (n = 10) root caries, using both a "most probable numbers" method and conventional plating methods. The predominant aciduric bacteria from root lesions were lactobacilli and A. israelii, while from sound root surfaces in subjects with root caries, A. gerencseriae comprised over 60% of aciduric isolates. Mutans streptococci were not among the aciduric isolates. Subjects without root caries harbored fewer bacteria, and S. anginosus (pH 4.8) and S. oralis (pH 5.2) were the predominant aciduric bacteria. The microbial etiology of root caries is more complex than was previously appreciated, and factors underlying the microbial succession occurring during the disease process are not known. Taxa with previously unrecognized aciduric characteristics have been isolated routinely, and the role of these organisms in the root caries process requires further investigation. PMID- 11926243 TI - Association of caries activity with the composition of dental plaque fluid. AB - This study tests the hypothesis that caries activity is associated with lower degrees of saturation with respect to enamel mineral in dental plaque fluid following sucrose exposure. Plaque fluids were obtained from caries-free, caries positive, and caries-active subjects. Samples were collected before and at 3 and 7 min after a sucrose rinse on consecutive weeks and analyzed for organic acids, inorganic ions, pH, calcium activity, and, in selected samples, total protein. After sucrose, pH values were significantly lower in the caries-active group in comparison with the caries-free and caries-positive groups. Total and free calcium concentrations increased with decreasing pH, with free calcium being about one-third of total calcium. The caries-active group exhibited significantly lower degrees of saturation with respect to enamel mineral, after sucrose, and had significantly higher mutans streptococci levels in plaque than did the caries free samples. Thus, saturation levels in post-sucrose plaque fluids reflect the cariogenic potential of dental plaque. PMID- 11926244 TI - Developmental changes in the nicotinic acetylcholine receptor in mouse tongue striated muscle. AB - There are no published studies on synaptogenesis focusing on the elimination of the superfluous nicotinic acetylcholine receptor (nAChR) outside the neuromuscular junction and the nAChR subunit switch from the embryonic-type (alpha2betagammadelta subunits) to the adult-type (alpha2betaepsilondelta subunits) in mouse tongues. To identify the time course of nAChR subunit elimination and switch, we analyzed the expression levels of alpha, epsilon, and gamma subunit mRNAs, and the immunolocalization of the delta subunit protein in the mouse tongue and corresponding hind limb. The analysis included the period from embryonic day (E) 11 to the newborn stage. The nAChR elimination and subunit switch began at E15 in the tongue and at E17 in the hind limb. They were nearly complete at birth in the tongue, but not in the hind limb. The early completion of synaptogenesis in the tongue at birth may be related to the early functional demands placed on the tongue, such as suckling and swallowing, immediately after birth. PMID- 11926245 TI - Maximum shortening velocity and myosin heavy-chain isoform expression in human masseter muscle fibers. AB - While human masseter muscle is known to have unusual co-expression of myosin heavy-chain proteins, cellular kinetics of individual fibers has not yet been tested. Here we examine if myosin heavy-chain protein content is closely correlated to fiber-shortening speed, as previously reported in other human muscles, or if these proteins do not correlate well to shortening speeds, as has been demonstrated previously in rat muscle. Slack-test recordings of single, skinned human masseter fibers at 15 degrees C revealed maximum shortening velocities generally slower and much more variable than those recorded in human limb muscle. The slowest fiber recorded had a maximum shortening velocity (V0) value of 0.027 muscle lengths x s(-1), several times slower than the slowest type I fibers previously measured in humans. By contrast, human limb muscle controls produced V0 measurements comparable with previously published results. Analysis by gel electrophoresis found 63% of masseter fibers to contain pure type I MyHC and the remainder to co-express mostly type I in various combinations with IIA and IIX isoforms. V0 in masseter fibers forms a continuum in which no clear relationship to MyHC isoform content is apparent. PMID- 11926246 TI - Mapping energy metabolism in jaw and tongue muscles during chewing. AB - Investigators have used positron emission tomography with 18F-fluoro-D deoxyglucose to obtain information not only for the diagnosis of cancers, but also for researching physiology in skeletal muscles. The aim of this study was to evaluate the activities of the jaw and tongue muscles during gum-chewing. Five volunteers aged 32-61 years were studied by positron emission tomography. They were requested to chew two pieces of chewing gum for 30 min after intravenous injection of 18F-fluoro-D-deoxyglucose. 18F-fluoro-D-deoxyglucose uptake in the intrinsic tongue muscle was significantly (p < 0.05) higher than that in the masseter, temporal, and medial pterygoid muscles. Heterogeneous uptake of 18F fluoro-D-deoxyglucose was observed in the masticatory muscles. In addition, the tongue exhibited higher activity than the masticatory muscles. In conclusion, positron emission tomography with 18F-fluoro-D-deoxyglucose appeared to be a useful technique for investigating the physiologic activities of the skeletal muscles, which have been difficult to examine by conventional methods. PMID- 11926247 TI - Activation of the bulbospinal serotonergic system during experimental tooth movement in the rat. AB - Experimental tooth movement is known to induce characteristic delayed and continuous nociception. Nociceptive somatic stimuli activate endogenous pain control systems such as descending monoaminergic pathways, which modulate the transmission of ascending sensory messages. To test the hypothesis that bulbospinal serotonergic pathways modulate subchronic nociception, we assayed the medulla at the level of the subnucleus caudalis and peri-aqueductal grey by high performance liquid chromatography with electrochemical detection for the serotonin (5-hydroxytryptamine, 5-HT) and its metabolite (5-hydroxyindoleacetic acid, 5-HIAA) 24 hrs after the onset of experimental tooth movement. Experimental tooth movement significantly increased 5-HT and 5-HIAA levels, and 5-HIAA/5-HT, an index of serotonin turnover, in the medulla, and 5-HIAA level and 5-HIAA/5-HT in the peri-aqueductal grey, indicating that nociception induced by experimental tooth movement activates the bulbospinal serotonergic pathway. PMID- 11926248 TI - Influences of ovariectomy on experimental tooth movement in the rat. AB - Estrogen withdrawal, which is important in the pathogenesis of post-menopausal osteoporosis, accelerates bone metabolism with a negative calcium balance. Therefore, it is hypothesized that estrogen deficiency could affect the rate of experimental tooth movement and alveolar bone remodeling. Six-week-old rats received a bilateral ovariectomy (OVX) or sham operation. Fourteen days later, rats were subjected to lateral tooth movement in the upper molar with nickel titanium wire of 10 g of force. OVX significantly increased the rate of experimental tooth movement from 12 days after experimental tooth movement (p < 0.001). Eighteen days after the start of tooth movement, bone histomorphometry demonstrated that OVX significantly elevated the osteoblast surface, osteoclast surface, and number of osteoclasts (p < 0.05) in the alveolar bone. These findings indicated that estrogen deficiency caused significantly rapid orthodontic tooth movement, and that the acceleration of tooth movement could be due to the further activation of alveolar bone turnover. PMID- 11926249 TI - Smokeless tobacco extracts modulate keratinocyte and fibroblast growth in organotypic culture. AB - Smokeless tobacco is associated with pathologic alterations of the oral mucosa, yet its direct effects on human keratinocytes and fibroblasts in stratified squamous epithelium are not well-understood. We hypothesized that smokeless tobacco could modulate the growth of keratinocytes and fibroblasts in an in vivo like, organotypic tissue model. To test this, we exposed organotypic cultures for 3 days to smokeless tobacco aqueous extracts and determined the changes in morphology and proliferation of human keratinocytes and fibroblasts. All smokeless tobaccos stimulated keratinocyte proliferation at low doses (0.25% w/v) and suppressed growth at higher doses (> 0.5% w/v). In contrast, smokeless tobacco extracts promoted fibroblast growth at all concentrations without inducing fibroblast turnover. Fibroblasts and keratinocytes, therefore, were differentially affected by smokeless tobacco extracts in an organotypic tissue model, suggesting incipient changes that may occur in vivo. PMID- 11926250 TI - Increasing awareness of danger signs in pregnancy through community- and clinic based education in Guatemala. AB - OBJECTIVE: This study evaluates the effectiveness of a set of information, education, and communication (IEC) strategies designed to increase the awareness of danger signs in pregnancy, delivery, or the postpartum period among pregnant or recently pregnant women. METHODS: Three IEC programs were implemented in 4 regions of southwestern Guatemala between April 1997 and May 1998: (1) a clinic based program involving the training of health providers in prenatal counseling and the provision of educational media to clients; (2) a community-based strategy consisting of radio messages regarding obstetric complications; and (3) educational sessions conducted through women's groups. Three surveys were conducted. In 1997, 637 pregnant women were interviewed at clinics where the interventions had been implemented. In 1998, 163 pregnant women using a subset of the same health clinics were interviewed. In 1999, a population-based survey of 638 pregnant and postpartum women was conducted. Using logistic regression, we model awareness of danger signs as a function of sociodemographic characteristics, prenatal care utilization, and IEC interventions. RESULTS: Among women using health clinics, the likelihood of having heard of danger signs nearly tripled between 1997 and 1998, when the clinic interventions were fully implemented. In 1999, those who had heard radio messages or participated in women's groups were, respectively, 3 times and 5 times more likely to have heard of danger signs in pregnancy. CONCLUSIONS: Safe motherhood programs can effectively increase knowledge of danger signs through clinic- and community based educational strategies. PMID- 11926251 TI - A comparison of risk factors for twin preterm birth in the United States between 1981-82 and 1996-97. AB - OBJECTIVE: This paper examines risk factors for twin preterm birth in 1981-82 and 1996-97 in the United States in order to see if they have changed over time. METHODS: We studied all U.S. twin births for the years examined (N = 346, 567). Since the gestational age distributions for twins differs from singletons, the risk of preterm birth was examined at <33, 33-34, and 35-36 weeks. Logistic regression was used to examine the contributions of sociodemographic and obstetric factors at each period. RESULTS: While the <33 week twin preterm rate rose 7% from 1981-82 to 1996-97, the 33-34-week rate rose 31%, and the 35-36-week rate rose 51%. Women with less education, teenagers, unmarried women, primiparas, and blacks were more likely to deliver preterm across all three preterm birth levels. However, the effect of these low socioeconomic status markers diminished over the study period. Additionally, the odds of preterm birth among blacks increased with earlier gestational ages. Women who had intensive prenatal care utilization as compared with less than adequate utilization were more likely to deliver preterm (35-36 weeks) in 1996-97 (odds ratio (OR) = 2.05) compared with 1981-82 (OR = 1.44). Smaller increases were noted for <33 and 33-34 weeks. CONCLUSIONS: Obstetric factors appear to be playing a greater role in the rise of twin preterm births at 35-36 weeks gestation. Temporal sociodemographic changes do not explain the rise in the preterm rate. Changing clinical practices may be having unintended consequences on the public health goals of reducing preterm and low birthweight rates in the United States. PMID- 11926252 TI - Pregnancy-associated obesity in black women in New York City. AB - OBJECTIVE: To determine weight gain during pregnancy and weight changes postpartum in first-time mothers delivering at or near term. METHODS: At about 2 weeks after delivery, 47 adult, Black and Hispanic women provided information on their prepregnancy weight and height and maximum pregnancy weight. Women reinterviewed at 2 and 6 months after delivery reported their most recent weight measurement and the date of that measurement. This information was used to compute each woman's prepregnancy body mass index, pregnancy weight gain, and weight loss postpartum. Information on infant feeding was also collected at each postpartum visit. RESULTS: About 2/3 of the women and 100% of the overweight and obese women gained excessive weight during pregnancy. Weight gain was most marked in women who started pregnancy overweight or obese. At 2 months postpartum, women were on average almost 18 lb above their prepregnancy weight. No additional maternal weight was lost by 6 months postpartum. Most infants were started on formula by 2 weeks of age. At 2 months of age, 85% were fed formula only and 91% of the infants were on WIC. CONCLUSIONS: Our results demonstrate a need for interventions to help women avoid obesity by regulating their pregnancy weight gain, losing weight for a longer period postpartum, and initiating and maintaining exclusive breast-feeding. PMID- 11926253 TI - The association between substance use and risky sexual behaviors among incarcerated adolescents. AB - OBJECTIVES: The purpose of this study is to examine the relationships between substance use and risky sexual behaviors and having acquired a sexually transmitted disease. METHODS: The sample consisted of 210 incarcerated adolescents ages 12-17. Data were collected through 40-min interviews administered by trained research assistants. Multivariate analysis was used to examine the relationship between risky sexual behaviors and substance use when adjusting for other covariates. RESULTS: Almost three-quarters (70.6%) of these incarcerated adolescents regularly use one or more substances. Inconsistent condom use and sex with multiple partners (>2) was reported by approximately 70% of those surveyed. The regular use of substances elevated the odds of having sex with multiple partners (OR = 11.88), exchanging sex for money or drugs (OR = 4.64). and inconsistent condom use (OR = 3.06). CONCLUSION: Given strong associations between risky sexual behavior and substance use, interventions should be more multiproblem-focused. Health interventions should attempt to address common causes of both behaviors. Effective interventions will be those that can successfully demonstrate effects on outcomes that measure both the intervention effects on substance use and risky sexual behaviors. PMID- 11926254 TI - Nonresponse in follow-back surveys of ethnic minority groups: an analysis of the Puerto Rican Maternal and Infant Health Study. AB - OBJECTIVES: This study documents the levels and sources of nonresponse in the first large-scale maternal-infant health survey administered to representative samples of Puerto Rican mothers on both the U.S. mainland and the island of Puerto Rico. METHODS: The data source is the Puerto Rican Maternal and Infant Health Study, which was administered to a vital records-based sample of 2763 mothers of infants. An additional 805 women were nonrespondents. Nonresponse is examined as a function of several characteristics measured from vital records using logistic regression. RESULTS: The response rate for this survey compares favorably to response rates for similar surveys. Although nonresponse is not associated with most characteristics measured from vital records, it is higher among mainland residents and mothers of infants who died. The absence of significant associations is due to opposite relationships between several covariates and the failure to locate and refusal. For example, nonresponse in the birth sample is not associated with migration, despite the difficulty of locating migrants. The lower likelihood of locating migrants is offset by their willingness to participate. CONCLUSIONS: Selectivity due to nonresponse is minimal. Nevertheless, researchers who design "binational" surveys should be aware of setting-specific circumstances that affect the ability to locate sampled individuals and secure their cooperation. PMID- 11926255 TI - Language proficiency and the enrollment of Medicaid-eligible children in publicly funded health insurance programs. AB - OBJECTIVES: The purpose of the study was to examine the effect of language proficiency on enrollment in a state-sponsored child health insurance program. METHODS: 1055 parents of Medicaid-eligible children, who were enrolled in a state sponsored child health insurance program, were surveyed about how they learned about the state program, how they enrolled their children in the program, and perceived barriers to Medicaid enrollment. We performed weighted chi2 tests to identify statistically significant differences in outcomes based on language. We conducted multivariate analyses to evaluate the independent effect of language controlling for demographic characteristics. RESULTS: Almost a third of families did not speak English in the home. These families, referred to as limited English proficiency families, were significantly more likely than English-proficient families to learn of the program from medical providers, to receive assistance with enrollment, and to receive this assistance from staff at medical sites as compared to the toll-free telephone information line. They were also more likely to identify barriers to Medicaid enrollment related to "know-how"--that is, knowing about the Medicaid program, if their child was eligible, and how to enroll. Differences based on language proficiency persisted after controlling for marital status, family composition, place of residence, length of enrollment, and employment status for almost all study outcomes. CONCLUSIONS: This study demonstrates the significant impact of English language proficiency on enrollment of Medicaid-eligible children in publicly funded health insurance programs. Strong state-level leadership is needed to develop an approach to outreach and enrollment that specifically addresses the needs of those with less English proficiency. PMID- 11926256 TI - Assessing the extent of medical home coverage among Medicaid-enrolled children. AB - OBJECTIVES: In light of the transition of the Alabama Medicaid program to a primary care case management model, we assessed the level to which children had access to a medical home before and after implementation of that model. Given the growing emphasis within the MCH community on assuring children medical homes, we explored whether Medicaid claims data could be used to assess medical home coverage. METHODS: We operationally defined "medical home" as use of a single primary care physician combined with receipt of at least one well child visit from that physician during the year. Using Alabama Medicaid claims data we assessed whether children's receipt of health care services met this defintion, the extent to which Medicaid-enrolled children had primary care providers and received well child visits, and changes in the source of well child visits before and after implementation of a primary care case management model in 26 of Alabama's 67 counties. RESULTS: In general, Medicaid-enrolled children in Alabama did not meet our definition of medical home either before or after implementation of a primary care case management model. Only 11.8% of children saw a single provider and had a well child visit from that provider during the baseline year. A majority of children (49.9%) however had both a primary care provider and received a well child visit. Sixteen percent of children saw a primary care physician but received no identifiable well visit, while 11% had well child care but did not see a primary care physician. Of particular concern, 23% neither saw a primary care physician nor had a well child visit during the baseline year. These figures changed only slightly in the 26 counties examined before and after implementation of the primary care case management model. CONCLUSIONS: State Maternal and Child Health programs are required to report as a performance measure "the percent of children with special health care needs in the state who have a medical/health home" as part of their Block Grant application. Using Medicaid data, this simple measurement strategy can provide an indication of the extent to which at least one population of children receive care through a medical home. PMID- 11926257 TI - Highlights from the National Summit on Safe Motherhood: Investing in the Health of Women. PMID- 11926258 TI - Snoezelen: an overview of research with people with developmental disabilities and dementia. AB - PURPOSE: This paper was to provide an overview of the research studies on snoezelen with people with developmental disabilities and dementia. METHOD: Computerized and manual searches were carried out to identify the aforementioned studies. Within-session, post-session, and longer-term effects of snoezelen were examined. RESULTS AND CONCLUSION: Twenty-one research studies were identified, 14 concerning people with developmental disabilities and seven people with dementia. Of those studies: 14 reported positive within-session effects; four positive post session effects; and two positive longer-term effects. These findings were discussed in relation to: (1) methodological aspects (weaknesses) of the studies; (2) the cost of arranging a snoezelen programme and possibilities of reducing the range of stimuli available in the programme; and (3) some research issues for advancing the understanding and effectiveness of intervention programmes with people with developmental disabilities and dementia. PMID- 11926259 TI - Understanding return to work behaviours: promoting the importance of individual perceptions in the study of return to work. AB - PURPOSE: The purpose of this paper is to demonstrate and discuss how individuals' subjective perceptions of personal and environmental issues influence return to work behaviour. METHOD: A qualitative design utilizing in-depth interviews and maximum variation sampling of 11 individuals who either returned to work or withdrew from work after a health leave was conducted. Experiences elicited were analysed using the constant comparative method followed by a member check with participants to confirm findings and interpretations. RESULTS: Findings underscored the importance of two key constructs in understanding return to work from the individual's perspective: the personal meaning of disability and return to work relevancy. Throughout the experience of getting better and returning to work participants reflected upon the impact of personal and external factors that contributed to their work disability, sought clarity of their performance capacities and examined the importance of work and the consequences of work disability within their life circumstances. CONCLUSIONS: Insights into an individual's perceptions of their impairment and the personal relevance of work can promote a better understanding of return to work behaviour. Integrating individual perceptions is essential to advancing a multidimensional approach in return to work research. PMID- 11926260 TI - Health-related quality of life after spinal cord injury. AB - PURPOSE: The purpose of this study was to assess health-related quality of life (HRQOL) in spinal-cord-injured adults living in Quebec (Canada). METHOD: Subjects with spinal cord injury (SCI) all of whom were members of the Quebec Paraplegic Association who gave their consent to participate completed the 36-item Short Form Health Survey (SF-36); sociodemographic and medical data were also solicited. RESULTS: Five hundred and eighty-seven participants were studied. The findings show a significant decrease in the score of the eight health concepts as measured by the SF-36 as well as that of the physical component summary measure (p<0.05). Analysing the various medical and sociodemographic variables with the eight scales of the SF-36 indicate that younger age, employment and the lack of hospitalization in the previous year were associated with a better quality of life. Mid- and long-term medical complications related to a spinal cord lesion and the impact of ageing are discussed briefly. CONCLUSION: HRQOL is decreased in the studied population with an SCI. PMID- 11926261 TI - Perception of Quality of Life by Chinese elderly persons with stroke. AB - PURPOSE: The purpose of this study was to initiate an investigation into the conceptualization of Quality of Life (QOL) for Chinese elderly people with stroke, living in the community in Hong Kong, and to explore whether a unique definition for this population might be needed. METHOD: Focus group interviews were conducted with seven elderly people with stroke and six healthy elderly individuals from community-based elderly centres in Hong Kong. A content analysis was conducted on interview data to determine the conceptual characteristics of QOL for participants with stroke, and to compare QOL components valued by both groups. RESULTS: Results appeared to support the assumptions in the literature about the multi-dimensionality of the QOL concept, its inclusion of both subjective and objective components, the interrelatedness of components and their integrated contribution to global QOL. The QOL components identified reflected a global QOL definition which included non-health-related life aspects such as money and housing. Similarities and differences were evident in the QOL components identified by both groups. These differences seemed to be best explained by the effects stroke had on the former group. CONCLUSIONS: The findings suggest that a QOL definition unique to Chinese elderly people with stroke may be warranted. Further research and refinement of the QOL conceptualization for this population are the necessary precursors to accurate QOL measurement and the planning and delivery of effective health services to help them achieve optimal QOL. PMID- 11926262 TI - Inter-observer reliability of clinical outcome measures in a lower limb amputee population. AB - PURPOSE: In an attempt to find a more clinically useful functional outcome measure specifically tailored for lower limb amputees undergoing inpatient prosthetic rehabilitation, a 6-month prospective assessment of inter-rater reliability for Harold Wood-Stanmore Mobility Scale Data, including two handicap scales, was undertaken. An analysis of the data is presented in this paper. METHODS: An inter-rater reliability study was undertaken using four observers to complete admission and discharge scores for the three disability/handicap scales on 14 consecutive patients over 6 months. RESULTS: The disability mobility scale demonstrated perfect observer agreement on admission and at discharge the inter rater reliability for this measure was high (0.83). By contrast, reliability between observers for admission scores on the handicap mobility scale was poor at 0.49 but reasonably high on discharge (0.83). On admission, inter-rater reliability for handicap physical independence was very low (0.15). At discharge, reliability improved to 0.69 being more consistent with results achieved for the other axes. CONCLUSIONS: This study confirms the good inter-rater reliability demonstrated previously in the literature but reveals poor inter-rater reliability for the two handicap scales. The latter will require modification before they can be used with confidence in conjunction with the disability scale. PMID- 11926263 TI - NS-417, a novel compound with neurotrophic-like effects. AB - NS-417 (5-(4-Chlorophenyl)-8-methyl-6-7-8-9-tetrahydro-1-H-pyrrolo[3.2 h]isoquinoline-2,3-dione-3-oxim hydrochloric acid salt) belongs to a new chemical series of compounds. NS-417 rescued differentiated PC12 cells from death induced by withdrawal of serum and nerve growth factor. Furthermore, NS-417 stimulated neurotrophic factor-induced neurite outgrowth in undifferentiated PC12 cells. In accordance with this observation, NS-417 potentiated NGF-induced signaling, such as activation of the extracellular signal-regulated kinases ERK1 and ERK2 and the Akt kinase. NS-417 also enhanced ERK activation induced by 10 minutes stimulation with NGF, bFGF or EGF in PC12 cells. In addition to the effect in PC12 cells, NS 417 increased the number of tyrosine hydroxylase (TH) positive cells in cultures established from dissociated E14 rat ventral mesencephali. PMID- 11926265 TI - Nuclear degradation of particular Fos family members expressed following injections of NMDA and kainate in murine hippocampus. AB - Transient glutamate signaling often leads to long lasting and permanent alterations of a variety of cellular functions through particular membrane receptors in the brain. For elucidation of mechanisms underlying long-term consolidation of transient extracellular signals, we have examined expression and degradation of particular Fos family member proteins required for assembly to the nuclear transcription factor activator protein-1 in this study. Transcription factors could modulate the activity of RNA polymerase II responsible for the formation of mRNA from genomic DNA in the nucleus and therefore regulate de novo synthesis of particular target functional proteins. Mice were intraperitoneally injected with 100 mg/kg N-methyl-D-aspartic acid (NMDA) or 40 mg/kg kainic acid (KA), followed by homogenization of hippocampus in the presence of different protease and phosphatase inhibitors 2 h after administration, and subsequent preparation of nuclear and cytosolic fractions. The systemic administration of both NMDA and KA induced marked expression of particular Fos family members, including c-Fos and Fra-2 proteins, in hippocampal nuclear and cytosolic fractions. Incubation at 30 degrees C for 1 to 18 h led to differential degradation profiles of each Fos family member protein in nuclear fractions in a manner peculiar to the individual excitants. Degradation rate was also affected by dialysis and subsequent addition of inhibitors for phosphatases and proteases. These results suggest that in vivo NMDA and KA signals may additionally modulate the activity of heterologous machineries responsible for breakdown of each Fos family member in a unique manner in nuclear fractions, rather than cytosolic fractions, of murine hippocampus. PMID- 11926264 TI - Mechanisms of action of carbamazepine and its derivatives, oxcarbazepine, BIA 2 093, and BIA 2-024. AB - Carbamazepine (CBZ) has been extensively used in the treatment of epilepsy, as well as in the treatment of neuropathic pain and affective disorders. However, the mechanisms of action of this drug are not completely elucidated and are still a matter of debate. Since CBZ is not very effective in some epileptic patients and may cause several adverse effects, several antiepileptic drugs have been developed by structural variation of CBZ, such as oxcarbazepine (OXC), which is used in the treatment of epilepsy since 1990. (S)-(-)-10-acetoxy-10,11-dihydro-5H dibenz [b,f]azepine-5-carboxamide (BIA 2-093) and 10,11-dihydro-10-hydroxyimino 5H-dibenz[b,f] azepine-5-carboxamide (BIA 2-024), which were recently developed by BIAL, are new putative antiepileptic drugs, with some improved properties. In this review, we will focus on the mechanisms of action of CBZ and its derivatives, OXC, BIA 2-093 and BIA 2-024. The available data indicate that the anticonvulsant efficacy of these AEDs is mainly due to the inhibition of sodium channel activity. PMID- 11926266 TI - Neurotoxicity of lindane and picrotoxin: neurochemical and electrophysiological correlates in the rat hippocampus in vivo. AB - In the present study, we compared in vivo changes of extracellular amino acid levels and nucleotide derivatives to a single ip dose of lindane (10-60 mg/kg) and picrotoxin (5 mg/kg) in the hippocampus of halothane anaesthetized rat by microdialysis-coupled HPLC analysis. Brain activity was monitored by EEG. The effects of lindane and picrotoxin on EEG pattern of rats as well as on hippocampal amino acid and nucleotide status were studied in 0-50 min, 50-100 min and 100-150 min periods post-dosing. Significant decreases in Glu and Asp were found after picrotoxin treatment. After 50-100 min post-dosing, hippocampal hypoxanthine and inosine levels increased to both lindane (10 mg/kg) and picrotoxin whereas xanthine and uridine levels increased to picrotoxin, only. Lindane elicited a dose-dependent occurrence of negative spikes accompanied with rhythmic activity at 4-5 Hz. The picrotoxin-induced 4-5 Hz activity did not display negative sharp waves and was accompanied by 10 Hz oscillations. PMID- 11926267 TI - Identification of transcripts expressed under functional differentiation in primary culture of cerebral cortical neurons. AB - In this study, we utilized primary cultures of cerebral cortical neurons and RNA arbitrarily primed polymerase chain reaction (RAP-PCR) to identify differentially expressed transcripts in neurons of different culture ages. Eleven cDNA fragments with high sequence similarity to known genes and Expressed Sequence Tags (ESTs) were cloned. From the National Center for Biotechnology Information (NCBI) sequence database, two clones were shown to be identical to known sequences, Mus musculus HP1-BP74 protein mRNA and Mus musculus KRAB-containing zinc finger protein, both were up-regulated. These genes have never before been shown to be involved in neuronal functional maturation. Among the remaining clones, clone 8 14 (239 bp) was very similar to Rattus norvegicus rS-Rex-b mRNA, which was further confirmed by sequencing its shortest isoform (1.5 kb) obtained by computer cloning. This study has identified eleven potential genes and transcripts, which might be involved in the development and differentiation of GABAergic neurons in culture. PMID- 11926268 TI - Taurine release in the developing and adult mouse hippocampus: involvement of cyclic guanosine monophosphate. AB - The inhibitory neuromodulator taurine is involved in osmoregulation and cell volume adjustments in the central nervous system. In addition, taurine protects neural cells from excitotoxicity and prevents harmful metabolic events evoked by cell-damaging conditions. The release of taurine in nervous cell preparations is greatly enhanced by glutamate receptor agonists and various cell-damaging conditions. NO-generating compounds also increase taurine release in the mouse hippocampus. The further involvement of the NO/cGMP pathway and protein kinases in preloaded [3H]taurine release from hippocampal slices from adult (3-month-old) and developing (7-day-old) mice in normoxia and in ischemia was now studied using a superfusion system. The release was enhanced by 8-Br-cGMP and the phosphodiesterase inhibitor 2-(2-propyloxyphenyl)-8-azapurin-6-one (zaprinast), particularly in the immature hippocampus, indicating that increased cGMP levels induce taurine release. The release was also increased by the inhibitor of soluble guanylyl cyclase, 1H-(1,2,4)oxadiazolo-(4,3a)quinoxalin-1-one (ODQ) and the protein kinase C activator 4beta-phorbol 12-myristate 13-acetate (PMA), but only in the adult hippocampus. The ischemia-induced release was also enhanced by increased cGMP levels in both adult and developing mice, whereas protein kinase inhibitors had no effects in any conditions. The results demonstrate that cGMP is able to modulate hippocampal taurine release in both adult and developing mice, the rise in cGMP levels evoking taurine release in normoxia and in ischemia. This could be part of the neuroprotective properties of taurine, being thus important particularly in cell-damaging conditions and in preventing excitotoxicity. PMID- 11926269 TI - Citrate, beneficial or deleterious in the CNS? AB - Cerebellar granule neurons were incubated with or without glucose (3 mM) in the presence or absence of citrate (20 mM) using normoxic and/or hypoxic incubation conditions. During 4 h of hypoglycemia and also during hypoxia plus hypoglycemia, citrate increased lactate dehydrogenase (LDH) leakage from the cells and decreased mitochondrial activity, the latter was also the case in the presence of glucose. After 24 h of hypoglycemia, however, citrate decreased LDH leakage slightly, possibly due to its metabolism in the tricarboxylic acid cycle under these conditions. It should be noted that during mild hypoxia plus hypoglycemia a reduced LDH leakage was observed when compared to hypoglycemia alone. The 4 h low oxygen period did protect the neurons also during the 20 h re-oxygenation period. The present study might indicate that incubation of brain cell cultures in an atmosphere of air (30% oxygen) and 5% CO2, which is used in most laboratories, can be toxic and that oxygen concentration should be lowered considerably to mimic conditions in the brain. PMID- 11926270 TI - Differential response of neural cells to trauma-induced free radical production in vitro. AB - CNS trauma has been associated with an increase in free radical production, but the cellular sources of this increase or the mechanism involved in the production of free radicals are not known. We, therefore, investigated the effects of trauma on free radical production in cultured neurons, astrocytes and BV-2 microglial cells. Free radicals were measured with the fluorescent dye DCFDA following in vitro trauma. At 30 and 60 min following trauma, there was a 132% and 64% increase, respectively, in free radical production in neurons when compared to controls. In astrocytes, there was a 94% and 133% increase at 30 and 60 min, respectively. Microglial cells, however, displayed no significant increase in free radicals at 30, 60 or 120 min following trauma. Since trauma can induce the mitochondrial permeability transition (MPT), a process associated with mitochondrial dysfunction, we further investigated whether cyclosporin A (CsA), an agent known to block the MPT, could prevent free radical formation following trauma. In neurons CsA did not block free radical production at 30 min but blocked it by 90% at 60 min. In contrast, in astrocytes CsA completely blocked free radical production at 30 min but did not block it at 60 min. Our results indicate that a differential sensitivity to trauma-induced free radical production exists in neural cells; that the MPT may be involved in the production of free radical post-trauma; and that the CsA-sensitive phase of free radical production is different in neurons and astrocytes. PMID- 11926271 TI - Effects of potassium and glutamine on metabolism of glucose in astrocytes. AB - The metabolic effects of extracellular glutamine (2.5 mM) or high potassium (25 mM) on glucose metabolism were studied in cultured cerebellar astrocytes. High potassium caused an increased glycolytic flux and an increase in glutamine release. Exposure to glutamine increased glycolytic flux and alanine formation, indicating that glutamine uptake is an energy requiring process. The effects of glutamine and high potassium on glycolytic flux were additive. Formation of metabolites from [1-13C]glucose and [2-13C]acetate confirmed the effects of glutamine and high potassium on glycolytic metabolism. In the presence of extracellular glutamine, analysis of the 13C labeling patterns of citrate and glutamine indicated a decrease in the cycling ratio and/or pyruvate carboxylation and glutamine synthesis from [1-13C]glucose did occur, but was decreased. Exposure to high potassium led to extracellular accumulation of acetate, presumably through non-enzymatic decarboxylation of pyruvate. PMID- 11926272 TI - The role of taurine in the central nervous system and the modulation of intracellular calcium homeostasis. AB - The effects of taurine in the mammalian nervous system are numerous and varied. There has been great difficulty in determining the specific targets of taurine action. The authors present a review of accepted taurine action and highlight recent discoveries regarding taurine and calcium homeostasis in neurons. In general there is a consensus that taurine is a powerful agent in regulating and reducing the intracellular calcium levels in neurons. After prolonged L-glutamate stimulation, neurons lose the ability to effectively regulate intracellular calcium. This condition can lead to acute swelling and lysis of the cell, or culminate in apoptosis. Under these conditions, significant amounts of taurine (mM range) are released from the excited neuron. This extracellular taurine acts to slow the influx of calcium into the cytosol through both transmembrane ion transporters and intracellular storage pools. Two specific targets of taurine action are discussed: Na(+)-Ca2+ exchangers, and metabotropic receptors mediating phospholipase-C. PMID- 11926273 TI - Effects of L-glutamate transport inhibition by a conformationally restricted glutamate analogue (2S,1'S,2'R)-2-(carboxycyclopropyl)glycine (L-CCG III) on metabolism in brain tissue in vitro analysed by NMR spectroscopy. AB - (2S,1'S,2'R)-2-(Carboxycyclopropyl)glycine (L-CCG III) was a substrate of Na(+) dependent glutamate transporters (GluT) in Xenopus laevis oocytes (IC50 to approximately 13 and to approximately 2 microM for, respec tively, EAAT 1 and EAAT 2) and caused an apparent inhibition of [3H]L-glutamate uptake in "mini slices" of guinea pig cerebral cortex (IC50 to approximately 12 microM). In slices (350 microM) of guinea pig cerebral cortex, 5 microM L-CCG III increased both the flux of label through pyruvate carboxylase and the fractional enrichment of glutamate, GABA, glutamine and lactate, but had no effect on total metabolite pool sizes. At 50 microM L-CCG III decreased incorporation of 13C from [3-13C] pyruvate into glutamate C4, glutamine C4, lactate C3 and alanine C3. The total metabolite pool sizes were also decreased with no change in the fractional enrichment. Furthermore, L-CCG III was accumulated in the tissue, probably via GluT. At lower concentration, L-CCG III would compete with L-glutamate for GluT and the changes probably reflect a compensation for the "missing" L-glutamate. At 50 microM, intracellular L-CCG III could reach > 10 mM and metabolism might be affected directly. PMID- 11926274 TI - Ammonia-induced extracellular accumulation of taurine in the rat striatum in vivo: role of ionotropic glutamate receptors. AB - Accumulation of taurine (Tau), glutamate (Glu) and glutamine (Gln) was measured in vivo in microdialysates of the rat striatum following a direct application to the microdialysis tube of 60 mM ammonium chloride which renders the final ammonia concentration in the extracellular space to approximately 5 mM. The following compounds were coadministered with ammonia to distinguish between the different mechanisms that may underlie the accumulation of amino acids: ion transport inhibitors, diisothiocyanostilbene-2,2'-disulfonate (DIDS) and furosemide, a Glu transport inhibitor L-trans-pyrrolidine-2,4-dicarboxylate (PDC), an NMDA receptor antagonist dizocilpine (MK-801) and an 2-amino-3-hydroxy-5-methyl-4 isoxazolepropionate (AMPA)/kainate (KA) receptor antagonist 6,7 dinitroquinoxaline-2,3-dione (DNQX). Ammonia stimulated Tau accumulation in the microdialysates to approximately 250% of the basal value. Furosemide did not significantly affect the stimulation by ammonia and DIDS only moderately depressed the effect. The ammonia-dependent Tau accumulation was increased by approximately 50% in the presence of PDC and reduced by approximately 35% in the presence dizocilpine and DNQX. In the microdialysates ammonia stimulated Glu and Gln accumulation somewhat less than Tau accumulation. Except for stimulation of Gln accumulation by DNQX, the effects were not modified by any of the cotreatments. The results are consistent with the assumption that ammonia stimulates Tau efflux mainly via activation of ionotropic Glu receptors. PMID- 11926275 TI - Metabolic compartmentation in cortical synaptosomes: influence of glucose and preferential incorporation of endogenous glutamate into GABA. AB - Metabolism of glutamine was determined under a variety of conditions to study compartmentation in cortical synaptosomes. The combined intracellular and extracellular amounts of [U-13C] GABA, [U-13C]glutamate and [U-13C]glutamine were the same in synaptosomes incubated with U-13C]glutamine in the presence and absence of glucose. However, the concentration of these amino acids was decreased in the latter group, demonstrating the requirement for glucose to maintain the size of neurotransmitter pools. In hypoglycemic synaptosomes more [U 13C]glutamine was converted to [U-13C]aspartate, and less glutamate was re synthesized from the tricarboxylic acid (TCA) cycle, suggesting use of the partial TCA cycle from alpha-ketoglutarate to oxaloacetate for energy. Compartmentation was studied in synaptosomes incubated with glucose plus labeled and unlabeled glutamine and glutamate. Incubation with [U-13C]glutamine plus unlabeled glutamate gave rise to [U-13C]GABA but not labeled aspartate; however, incubation with [U-13C]glutamate plus unlabeled glutamine gave rise to [U 13C]aspartate, but not labeled GABA. Thus the endogenous glutamate formed via glutaminase in synaptic terminals is preferentially used for GABA synthesis, and is metabolized differently than glutamate taken up from the extracellular milieu. PMID- 11926276 TI - Evaluation of drugs acting at glutamate transporters in organotypic hippocampal cultures: new evidence on substrates and blockers in excitotoxicity. AB - Removal of L-glutamate (Glu) from the synapse is critical to maintain normal transmission and to prevent excitotoxicity, and is performed exclusively by excitatory amino acid transporters (EAATs). We investigated the effects of substrates and blockers of EAATs on extracellular Glu and cellular viability in organotypic cultures of rat hippocampus. Seven-day treatment with a range of drugs (L-trans-pyrrolidine-2,4-dicarboxylate, (2S,4R)-4-methyl-glutamate, (+/-) threo-3-methylglutamate and DL-threo-beta-benzyloxyaspartate), in the presence of 300 microM added Glu, resulted in increased extracellular Glu and a significant correlation between Glu concentration and cellular injury (as indicated by lactate dehydrogenase release). In contrast, (2S,3s,4R)-2 (carboxycyclopropyl)glycine (L-CCG-III) exerted a novel neuroprotection against this toxicity, and elevations in extracellular Glu were not toxic in the presence of this compound. Similar results were obtained following two-week treatment of cultures without added Glu. Whilst blockade of GLT-1 alone was relatively ineffective in producing excitotoxic injury, heteroexchange of Glu by EAAT substrates may exacerbate excitotoxicity. PMID- 11926277 TI - Osmosensitive release of neurotransmitter amino acids: relevance and mechanisms. AB - Hyposmolarity activates amino acid efflux as part of the corrective volume process in a variety of cells. This review discusses the mechanism of amino acid release in brain cells preparations. Results present evidence of substantial differences between the efflux of taurine and that of GABA and glutamate, which besides a possible role as osmolytes, have a main function as synaptic transmitters. The differences found concern the efflux time course, the sensitivity to C1- channel blockers, the modulation by tyrosine kinases, the influence of PKC and the effect of cytoskeleton disruptive agents. While taurine efflux features fit well with the mechanisms so far described in most cell types, the efflux of GABA and glutamate does not. Alternate mechanisms for the release of these two amino acids are discussed, including a PKC-modulated, actin dependent exocytosis. PMID- 11926278 TI - Differential mechanisms of glutamate-stimulated perturbations in the kinetics of c-fos mRNA induction are associated with maturation of cerebellar granule cells in primary culture. AB - In further exploring proposals for the measurement of early gene (c-fos mRNA) levels as a predictive index for in vitro excitotoxicity, this study, using immature (2 days in vitro) cultures of mouse cerebellar granule cells as an experimental model system, was undertaken to determine the effect of glutamate (Glu) i) in stimulating increases in intracellular free-calcium ([Ca2+]), ii) on cell viability and iii) on induction of steady-state c-fos mRNA levels. In parallel experiments the action of agents (viz. 55 mM KCl and the calcium ionophore, A23187) that mediate Ca2+ entry into cells via different routes was also evaluated. Glu was unable to induce excitotoxicity in granule cells at this stage of development in culture, but did stimulate a concentration-dependent and marked increase in [Ca2+], levels while also mediating a dramatic concentration dependent perturbation in the kinetics of c-fos mRNA induction that appeared to arise solely from NMDA receptor-mediated Ca2+ influx. The results are presented in comparison to the actions of KCI and A23187 and considered in relation to earlier studies undertaken using mature (7 days in vitro) cultures of cerebellar granule cells. PMID- 11926280 TI - Immunogold cytochemistry identifies specialized membrane domains for monocarboxylate transport in the central nervous system. AB - An efficient exchange of lactate between different cell types (such as astrocytes and neurones) would require that lactate transporters are expressed in contiguous parts of the respective plasma membranes. To settle this issue we explored the subcellular expression pattern of monocarboxylate transporters (MCTs) by use of selective antibodies and high resolution immunogold cytochemistry. We investigated whether the membrane domains containing MCT1, MCT2 and MCT4 are spatially related to each other and to other membrane domains, i.e. those containing glutamate receptors. We used retina and cerebellum as a model for our investigations. We found that MCT1 was localized in the apical membrane of pigment epithelial cells and in the photoreceptor inner segment membrane in the retina. In the brain MCT1 was present in endothelial cells. MCT2 was localized in the postsynaptic membrane of parallel fiber-Purkinje cell synapses and MCT4 was situated in the membrane of glial cells in the cerebellum. PMID- 11926279 TI - Glutamate-induced excitotoxicity in retina: neuroprotection with receptor antagonist, dextromethorphan, but not with calcium channel blockers. AB - The purpose of our studies was to evaluate different strategies for possible neuroprotection in glutamate-induced neurotoxicity in the retina. In a first set of experiments we attempted to determine if dextrorphan antagonism of glutamate action on NMDA receptors would protect against excitotoxic injury associated with secondary damage seen after surgical laser treatment in retina. In a second set of experiments, the effects of different calcium channel blockers in an in-vitro model of N-methyl-D-aspartate (NMDA)-induced retinal ganglion cell excitotoxicity that utilized rabbit retinal explants were evaluated. Dextrorphan infusion prior to laser treatment of rabbit retina produced a significant decrease in the area of neural retinal damage. We attribute the apparent dextrorphan protection to attenuation of glutamate mediated excitotoxicity secondary to laser induced cell death. Preincubation of rabbit retinal explants with verapamil, nimodipine or omega-conotoxin MVIIA did not cause a significant change in NMDA induced cell death in the ganglion cell layer. PMID- 11926282 TI - Nitrofurazone induces non-regenerative hepatocyte proliferation in rats. AB - The antibiotic nitrofurazone (NF) has been known for its testicular toxicity; in contrast, much less is known about its effect on the liver. NF was given to male rats for up to 7 consecutive days to evaluate NF-induced effects on the liver. NF increased hepatocyte DNA synthesis and liver weight in a dose-dependent manner, with no apparent histological or biochemical evidence of cell damage or loss. The hepatocyte proliferation ceased after a few days despite the continuation of treatment. The absence of cell damage indicates that NF-induced hepatocyte proliferation is different from regenerative proliferation that is seen after partial hepatectomy or cell necrosis. PMID- 11926283 TI - In contrast to neuronal NOS-I, the inducible NOS-II expression in aging brains is modifled by enriched environmental conditions. AB - Changes in the expression of the nitric oxide synthase isoforms (NOS) have been implicated in age-related neurodegeneration, although mechanisms responsible for specific differences such as reduced cognitive and motor functions are not well understood. Immunohistochemical examination of the age-dependent NOS-I and NOS-II expression in aging rats at 3, 12, 24, 36 months of age as well as in 36 months old rats who spent their last 3 months in an enriched environment, revealed a significant decrease in NOS-I expression with aging. In contrast, NOS-II increased at 36 months. Old rats kept under enriched environmental conditions, however, showed strongly reduced NOS-II expression. These results indicate that both NOS-I and II may contribute to age-related degenerative processes, but in contrast to NOS-I the age-dependent changes of NOS-II are reduced or even reversed by stimulating environmental conditions. PMID- 11926284 TI - Role of exogenous melatonin in reducing the cardiotoxic effect of daunorubicin and doxorubicin in the rat. AB - The aim of the studies was to examine the cardioprotective effect of melatonin during the anthracycline administration (daunorubicin, doxorubicin) in rats. Application of these drugs in chemotherapy is limited because of their cardiotoxicity. Rats of Buffalo strain were divided into groups according to the cytostatic drug used, its dose and sequence of administration (single intravenous [i.v.] dose of 10 mg/kg b.w., i.e., acute intoxication; 3 mg/kg b.w. weekly for 3 weeks, subchronic intoxication). Melatonin was administered subcutaneously before and after every injection of a cytostatic drug at a dose of 10 mg/kg b.w. The degree of cardiac muscle cell alterations was examined either histologically (Mean Total Score technique and the Billingham scale), or biochemically (levels of lipid peroxidation markers, malonyldialdehyde, and 4-hydroxyalkenals). Statistically significant decrease in cardiac muscle cell damage was noted with an aid of the Billingham scale after melatonin administration in acutely intoxicated doxorubicin-treated rats (p < 0.001). The similar phenomenon was observed using the Mean Total Score technique in case of acute daunorubicin or doxorubicin (p < 0.01 and p < 0.001, respectively) intoxications. A significant reduction in cardiac muscle cell lesions was detected either by the Billingham scale or by the Mean Total Score technique during subchronic intoxication with either of the anthracyclines when melatonin was given. Biochemical assays revealed significant decreases in malonyldialdehyde and 4-hydroxyalkenals levels following application of melatonin during either acute doxorubicin (p < 0.05) or subchronic daunorubicin (p < 0.01) intoxication. In summary, melatonin was found to exert a protective effect on the cardiac muscle cells, which was particularly evident after acute doxorubicin or subchronic daunorubicin intoxication, using either histological or biochemical methods. PMID- 11926281 TI - Delayed cell death signaling in traumatized central nervous system: hypoxia. AB - There are two different ways for cells to die: necrosis and apoptosis. Cell death has traditionally been described as necrotic or apoptotic based on morphological criteria. There are controversy about the respective roles of apoptosis and necrosis in cell death resulting from trauma to the central nervous system (CNS). An evaluation of work published since 1997 in which electron microscopy was applied to ascertain the role of apoptosis and necrosis in: spinal cord injury, stroke, and hypoxia/ischemia (H/I) showed evidence for necrosis and apoptosis based on DNA degradation, presence of histones in cytoplasm, and morphological evidence in spinal cord. In the aftermath of stroke, many of the biochemical markers for apoptosis were present but the morphological determinations suggested that necrosis is the major source of post-traumatic cell death. This was not the case in H/I where both biochemical assays and the morphological studies gave more consistent results in a manner similar to the spinal cord injury studies. After H/I, major factors affecting cell death outcomes are DNA damage and repair processes, expression of bcl-like gene products and inflammation-triggered cytokine production. PMID- 11926285 TI - Nivalenol--induced apoptosis in thymus, spleen and Peyer's patches of mice. AB - ICR:CD-1 male mice were orally administered with Nivalenol(NIV) at the dose levels of 5, 10 and 15 mg/kg body weight, and examined at 12, 24 and 48 hours after inoculation (HAI), respectively, to elucidate the process of development of apoptosis in the thymus, spleen and Peyer's patch. There were no signs of clinical disorders and no changes in body and organ weights until 48 HAI except for that the thymus weight significantly decreased at 48 HAI. Immunohistochemically, the number of apoptotic lymphocytes evaluated by in situ detection for fragmented DNA showed a dose-dependent increase at 12 HAI in both the thymus and the Peyer's patch, while it became to increase at 24 HAI in the spleen. Dead lymphocytes in the thymus, spleen and Peyer's patch showed ultrastructural characteristics of apoptosis. Moreover, the DNA ladder was first detected by agarose gel electrophoresis at 12 HAI in the thymus of 15 mg/kg group. The results clearly indicate that NIV is able to induce apoptosis in the lymphoid tissues of mice. PMID- 11926286 TI - Role of endotoxin in 6-sulfanilamidoindazole(6SAI)-induced arthritis in rats. AB - 6-Sulfanilamidoindazole (6SAI) induces selflimiting arthritis in rats. Since close relationships exist between arthritis and endotoxin, four experiments were conducted to clarify the relationship between endotoxin and 6SAI-induced arthritis. Endotoxin levels in the plasma from the abdominal aorta and portal vein from rats that had 6SAI (500 mg/kg) administered orally for up to 7 days remained within the control values at day 1 and day 3, and were significantly elevated at day 7. Endotoxin levels in the synovial fluid from the same rats showed no significant change. Ankle swelling and redness in rats treated 11 consecutive days with 6SAI did not ameliorate when coadministered with an anti endotoxin agent, polymyxin B sulfate. Histopathological examination on the ankles of rats treated orally with non-arthiritogenic sulfonamides including sulfonamide, sulfamethoxazole and sulfadimethoxin (250 and 500 mg/kg/day, each compound) for 2 weeks demonstrated no inflammatory changes, while hyperplasia/hypertrophy of thyroid epithelial cells were frequently observed. When histopathological changes in the ankles from rats coadministered with 6SAI and lipopolysaccharide (LPS, Escherihia coli O55:B5, 50 microg/kg, i.v.) were compared with those in rats treated with 6SAI or LPS alone, the ankles from the 6SAI+LPS treated animals had marked edematous inflammation in the synovium and surrounding connective tissues, whereas the LPS-group had only mild focal infiltration of polymorphonuclear leukocytes in the synovium and the 6SAI-group showed no apparent changes. These results suggest that endotoxin is not a direct cause but a possible acceralating factor of 6SAI-induced arthritis, and that the effects of 6SAI on gut bacteria is not related with the pathogenesis of this model. PMID- 11926287 TI - Hepatic lysosomal enzymes activity and liver morphology after short-time omeprazole administration. AB - The aim of the study was to establish the influence of short-time omeprazole administration on liver function and morphology. Omeprazole was administered intraperitoneally, twice daily, for 3 days to male Wistar rats in two doses: 0.571 mg/kg and 5.71 mg/kg. Control animals were treated with physiological saline. Half of the animals were sacrificed 12 hours after the last injection. The remaining rats were raised for another 6 weeks, without any xenobiotics, and sacrificed on the 47th day of the experiment. The activity of free and bound fractions of hepatic acid phosphatase, beta-galactosidase, beta-N-acetyl glucosaminidase, cathepsin B, D and L, lipase, and sulphatase were determined spectrophotometrically in homogenates of the liver. The liver sections were examined by light microscopy with hematoxylin-eosin, azan, and periodic acid Schiff stains. Marginally significant (p < 0.1) differences in activity of free sulphatase fraction, and free and bound fractions of beta-galactosidase were found in animals exposed to the higher dose of omeprazole and sacrificed 12 hours after the last injection. Enzymatic profiles were normalised during the next 6 weeks. Histological evaluation revealed small degenerative and adaptive changes in all examined groups. It could be concluded that observed differences of hepatic lysosomal enzyme activities were the result of accompanied chemical induced peritonitis as previously reported, and not a direct drug-toxic effect. PMID- 11926289 TI - Possible role of 1-methylnicotinamide in the pathogenesis of Parkinson's disease. AB - This study tested the hypothesis, that nicotinamide N-methyltransferase (NAMT) activity in the brain could convert nicotinamide to 1-methylnicotinamide (MNA) and by that means damage the nigro-neostriatal dopaminergic neurons. The NAMT activities of rat brain and liver were assayed with gas chromatographic-mass spectrometric analysis in a selected ion monitoring system. They amounted to 0.30 nmol/mg x h and 0.51 nmol/mg x h, respectively. The MNA injection in rat substantia nigra pars compacta significantly decreased dopamine content in the striatum. NADH oxidation and lipid peroxidation by MNA via rat brain submitochondrial particles (SMP) under the condition of pH ranging from pH 6.0 to 10.0 were verified. The pH optimum for the NADH oxidation was 9.0. The pH optimum for the peroxidation of the lipid composing SMP by MNA was also 9.0. The lipid peroxidation in this assay was suppressed by superoxide dismutase. The superoxide anion formed by MNA via mitochondria might be involved in the etiology of Parkinson's disease. PMID- 11926288 TI - Transplacental effects of 2,3,7,8-tetrachlorodibenzo-p-dioxin on the temporal modulation of Sp1 DNA binding in the developing cerebral cortex and cerebellum. AB - Gestational exposure to environmental pollutants such as 2,3,7,8 tetrachlorodibenzo-p-dioxin, (dioxin) often leads to neurodevelopmental and neurobehavioral deficits, which clearly suggest the involvement of the central nervous system. The objective of this study was to determine the effects of an acute, gestational exposure to dioxin on the developmental expression profile of a transcription factor (Sp1) that is involved in growth and differentiation in the developing brain of F, generation pups. Timed-pregnant Harlan Sprague Dawley rats were exposed to single oral doses of 0.25, 0.5, and 1.0 microg dioxin/kg body weight or vehicle corn oil on gestation day 15. Electrophoretic mobility shift assays (EMSA) of the nuclear extracts from the cerebral cortex and cerebellum of the resulting pups, on postnatal days (PND) 3.5, 10, 15, 20 and 30, with an Sp1 consensus oligonucleotide sequence revealed modulations in the DNA binding activity of Sp1 as a result of the transplacental dioxin exposure. In the cerebral cortex, a dose-dependent premature peak of Sp1 DNA-binding on PND 3 was observed when compared with the control animals where Sp1 DNA-binding was maximal on PND 10. In the postnatally-developing cerebellum, a dose-dependent premature peak of Sp1 DNA binding on PND 5 was observed when compared with the control animals where Spl DNA-binding was maximal on PND 15. The mean birth index, body weight gain, and gross brain weight of the pups were not statistically different from the control animals over the 30 day postnatal period studied. The data obtained on the Sp1 developmental expression profiles in the brain indicate that (1) Sp I DNA-binding is developmentally regulated and expressed very highly in actively developing brain regions, and (2) a potential consequence of the transplacental effect of dioxin to the fetus is in utero neurotoxicity. PMID- 11926290 TI - Ultrastructural changes in the dorsal skin of Wistar-derived hypotrichotic WBN/ILA-Ht rats exposed to subchronic ultraviolet B (UVB)-irradiation. AB - Ultrastructural changes in the dorsal skin were examined in Wistar-derived hypotrichotic WBN/ILA-Ht rats exposed to subchronic UVB-irradiation (10 kJ/m2 per rat per day for up to 3 months). Epidermal hyperplasia developed at I month of UVB-irradiation and progressed thereafter, resulting in epidermal thickening and formation of epidermal ingrowths projecting into the dermis. In some portions of the epidermal ingrowths at 2 and 3 months, keratinocytes were somewhat pleomorphic. In addition, some of the keratinocytes showing cytoplasmic projections migrated into the dermis. The basement membrane and hemidesmosomes at the epidermal-dermal junction became to disappear along with the development of edema spreading from the upper dermis to the epidermis. However, Langerhans cells were still detected in the hyperplastic epidermis even at 3 months. In the dermis, in addition to edema, fibroblast proliferation and mast cell infiltration progressed with time, and degranulation of mast cells was obvious at 2 and 3 months. Only a few basophils as well as eosinophils were also found. In the upper dermis, especially beneath the epidermis, decrease in diameter and disintegration of collagen fibrils were observed. Ultrastructural characteristics of the dorsal skin responses to subchronic UVB-irradiation were clarified in the present study. PMID- 11926291 TI - Experimental one year ochratoxin A toxicosis in pigs. AB - Mild mycotoxic nephropathy was induced in 6 pigs by a diet containing ochratoxin A at 800 ppb, several times higher than that naturally encountered in some feed for pig production in Bulgaria. The nephropathy was expressed only as slightly hypertrophied kidneys with a faintly mottled surface, discernible at the end of the experiment to a skilled observer but probably not recognisable in routine slaughterhouse processing. Histological examination showed two types of changes: degenerative - affecting epithelial cells in some proximal tubules of pigs after 6 months, and proliferative changes in the interstitium which predominated after 1 year of exposure to ochratoxin A. Telangiectasis and lymph stasis were rarely seen. The renal lesions were similar to those described for classical mycotoxic porcine nephropathy formerly encountered in Denmark, but they were rather different from the porcine nephropathy which occurs spontaneously in Bulgaria. Measurement of ochratoxin A in serum provided analytical values complementary to feed intake and with similar concentration values. It also showed both accumulation with time, from 3 months to 6 months (approximately 1 ppm), and a 2 fold range of values within a group eating from a common feed source, as in commercial pig production. Mild symptomatology in this long, single-mycotoxin experiment serves to lessen somewhat the current perception of the direct renal toxicity of ochratoxin A alone, though a role in multi-toxin contexts is unquestioned. PMID- 11926293 TI - Granulocyte colony-stimulating factor exacerbates the acute lung injury and pulmonary fibrosis induced by intratracheal administration of bleomycin in rats. AB - We investigated the effects of granulocyte colony-stimulating factor (G-CSF) on lung injury induced by intratracheal administration of bleomycin (BLM, 2 mg/200 micro1) in rats. In experiment 1, G-CSF (10, 30 and 100 microg/kg/day, s.c.) was administered to rats treated with BLM or saline for 7 days starting immediately after BLM administration. In rats receiving G-CSF alone, a large number of neutrophils were noted in the pulmonary capillaries, although there were no lung lesions. In rats receiving BLM alone, diffuse alveolar damage was observed. The administration of G-CSF to BLM-treated rats increased the total lung lesion per unit of pulmonary parenchyma (total lung lesion %) along with increases in the peripheral neutrophil count and the number of neutrophils infiltrating in the pulmonary lesion in a dose-dependent fashion. In experiment 2, 100 microg/kg/day of G-CSF was administered to rats treated with BLM or saline for up to 28 days starting immediately after BLM administration. The administration of 100 microg/kg/day of G-CSF to BLM-treated rats showed no effects at 14 days but it increased the lung lesion % and the score of lung fibrosis along with the increase in the number of neutrophils infiltrating in the pulmonary lesion at 28 days. These findings suggest that G-CSF administration to BLM-treated rats influenced and exacerbated the BLM-induced acute lung injury, and also exacerbated pulmonary fibrosis in a dose-dependent fashion. The exacerbation of lung injury coincided with the marked increase in the peripheral neutrophil count and the number of neutrophils infiltrating in the pulmonary lesion. PMID- 11926294 TI - The perils of appeasement. PMID- 11926292 TI - Acetaminophen hepatotoxicity and mechanisms of its protection by N acetylcysteine: a study of Hep3B cells. AB - Acetaminophen (AAP) hepatotoxicity, resulting in centrilobular necrosis, is frequently encountered following suicidal attempts, especially by adolescents, but also after its excessive use in infants. The subcellular and molecular sequences leading to hepatocellular cell death are not yet clear. We therefore investigated AAP hepatotoxicity by using cultured hepatoma-derived cells (Hep3B) exposed to AAP and N-acetylcysteine (NAC), used as a protective agent. Specifically, we studied the role of apoptosis and oxidative damage as putative mechanisms of AAP-associated cytotoxicity. Hep3B cells were exposed to AAP (5-25 mM) and NAC (5 mM) for different time periods. Cell viability was assessed by the Alamar Blue Reduction Test and LDH. Oxidative damage was evaluated by measuring reactive oxygen species (ROS) and glutathione. AAP-induced apoptosis was investigated by flow cytometry and transmission electron microscopy. We found that: 1. In Hep3B cells, AAP causes a time- and concentration-dependent cytotoxic effect, leading to oxidative stress, mitochondrial dysfunction, alterations of membrane permeability and apoptosis; 2. In the course of AAP cytotoxicity, the generation of ROS appears as an early event which precedes decrease of viability, LDH leakage, glutathione depletion and apoptosis; 3. NAC protects Hep3B cells from AAP-induced oxidative injury, but does not prevent apoptosis. PMID- 11926295 TI - Non-invasive measurement of blood pressures in the Yucatan micropig (Sus scrofa domestica), with and without midazolam-induced sedation. AB - Current literature suggests that the effects of midazolam, a water-soluble benzodiazepine, on blood pressure in swine are minimal. The hypothesis of the study reported here was that a light sedative dose would induce a decrease in blood pressure in this species. Healthy female Yucatan Micropigs (n = 20), 16 to 30 (mean, 22) kg, aged four six months, were individually placed in a humane restraint sling and allowed to acclimate. Systolic (SBP), diastolic (DBP), and mean (MBP) blood pressures (mmHg) and heart rate (HR; beats per min [bpm]) were measured by use of oscillometry. The pressure cuff was placed at the base of the tail, and five sets of values were recorded at five-min intervals, beginning at 10 and ending 30 min after cuff placement. Following a three- to four-day rest period, this procedure was repeated with the addition of a dose of 0.5 mg of midazolam HCl/kg of body weight given intramuscularly at the time of cuff placement. A paired one-way Student's t-test was used to compare the means of the five measures between control and midazolam treatment. Mean (+/- SD) differences for SBP, DBP, MBP, and HR were 18.9 (+/- 3.97), 17.8 (+/- 5.27), and 18.6 (+/- 5.09) mmHg and 20.7 (+/- 3.73) bpm, respectively. All four parameters were significantly reduced in the midazolam-sedated group (P < 0.001). The maximal decrease in SBP, DBP, and MBP occurred at 15 and 20 min after dosing. Mean values based on the means of the five measures were 128 (+/- 12.6), 80 (+/- 9.4), and 99 (+/- 9.2) mmHg and 135 (+/- 17.4) bpm, and 109 (+/- 15.4), 63 (+/- 12.6), and 80 (+/- 13.6) mmHg and 115 (+/- 15.5) bpm for SBP, DBP, MBP, and HR in the control (n = 20) and midazolam (n = 20) groups, respectively. The control values can serve as normal oscillometric values for this age, sex, and breed of Micropig. We conclude that midazolam, given intramuscularly at a sedative dosage, negatively affects cardiovascular parameters measured by use of a blood pressure cuff, in sexually mature female Micropigs, compared with values in untreated pigs, which is similar to reports for humans. PMID- 11926297 TI - Anomalous (preduodenal) portal vein: autosomal recessive mutation in AKR/J mice. AB - BACKGROUND AND PURPOSE: Anomalous (preduodenal) portal vein was found in AKR/J mice. It is a rare congenital malformation in humans, and to the authors' knowledge, has never been reported in laboratory animals. Morphology, clinical signs of disease, and heritability of this anomaly were examined. METHODS: Fifty three strains of inbred mice (6,026 mice) in our mouse colony were examined for preduodenal portal vein and its association with clinical signs of disease (vomiting or abdominal pain) and other anomalies. Heritability also was tested by use of cross-backcross matings of AKR/J mice with clinically normal PT mice. RESULTS: The portal vein was found at the ventral side of the duodenum in most (98%) AKR/J mice, whereas it ran at the dorsal side of the duodenum in 52 other inbred mouse strains in our mouse colony. Clinical signs of disease and other congenital anomalies were not detected in this strain of mice, although position has a high association with other congenital anomalies in humans. Heritability testing of this anomaly in AKR/J mice indicated single autosomal recessive inheritance. CONCLUSIONS: Preduodenal portal vein found in AKR/J mice is a single autosomal recessive mutation, but is not associated with clinical signs of disease and other congenital malformations. PMID- 11926296 TI - Effects of inhibitors on chicken polymorphonuclear leukocyte oxygenation activity measured by use of selective chemiluminigenic substrates. AB - Chicken heterophil polymorphonuclear leukocytes (CPMNLs) have NADPH oxidase activity, but lack myeloperoxidase (MPO). Stimulation of CPMNLs by phorbol 12 myristate 13-acetate or chicken opsonified zymosan results in luminol-dependent chemiluminescence (CL) activity, which is small relative to that of human peroxidase-positive neutrophils (HPMNLs), as well as lucigenin-dependent CL, comparable to HPMNL responses. Inhibitors were used to investigate and characterize the CL activity of CPMNLs. Inhibition constants were calculated, using Dixon inhibition analysis, or were reported as the concentration producing 50% inhibition of the magnitude of CL responses. Azide and cyanide are effective inhibitors of luminol CL in HPMNLs, although these peroxidase inhibitors do not inhibit either luminol or lucigenin CL of CPMNLs. Since these agents also inhibit eosinophil peroxidase, lack of inhibition of CPMNL CL indicates that the small percentages of peroxidase-positive eosinophils in CPMNL preparations are not responsible for the luminol CL observed. Iodoacetate and fluoride, pre-oxidase and pre-peroxidase inhibitors of glycolytic metabolism, effectively inhibit lucigenin and luminol CL activities in CPMNLs. Superoxide dismutase competitively inhibits lucigenin and luminol CL in CPMNLs, but catalase is an ineffective inhibitor. Although luminol is efficiently dioxygenated by a MPO-dependent mechanism in HPMNL, use of peroxidase-deficient CPMNLs indicates that this substrate does not exclusively measure peroxidase activity. PMID- 11926298 TI - Effects of tribromoethanol anesthesia on echocardiographic assessment of left ventricular function in mice. AB - BACKGROUND AND PURPOSE: Pentobarbital and ketamine-xylazine anesthesia in mice result in markedly decreased left ventricular fractional shortening and cardiac output. However, to the authors' knowledge, the effect of short-acting, alcohol based anesthesia on these parameters is unknown. METHODS: Fifteen mice (FVB/N, C57Bl/6J, A/J, n = 5 each) underwent high-resolution (15 MHz) 2-dimensional directed M-mode echocardiography before and after undergoing 2.5% tribromoethanol anesthesia (0.01 ml/g of body weight). RESULTS: Tribromoethanol anesthesia resulted in significant heart rate slowing (29%) and left ventricular enlargement (20%), and a more modest (12%) reduction in left ventricular fractional shortening. Cardiac output was unchanged. The differences in left ventricular function between conscious and tribromoethanol studies were similar for each of the three strains of mice. CONCLUSIONS: Tribromoethanol anesthesia induced only modest effects on M-mode estimates of basal cardiac function and did not influence cardiac output. The effects to tribromoethanol anesthesia were similar among three commonly used mice strains. PMID- 11926299 TI - Hematologic and serum biochemical reference intervals for the chimpanzee (Pan troglodytes) categorized by age and sex. AB - Normal reference range intervals for hematologic and serum biochemical values in the chimpanzee (Pan troglodytes) have seldom been reported. The few studies that have been conducted either report values on the basis of a small number of animals, report values for all age groups or both sexes combined, or were designed specifically to document the effect of a particular condition on the normal range of hematologic and serum biochemical values. On the basis of data collected from 133 chimpanzees over a 17-year period, empirically based clinical reference ranges were derived to provide a guide for basic diagnostic and clinical care of chimpanzees. For either sex within each of four age groups, there is a table that summarizes serum biochemical and a table that summarizes hematologic values. These values are compared with prior values, and their importance in the care and well being of captive chimpanzee populations is discussed. PMID- 11926300 TI - Prevalence of rat virus infection in progeny of acutely or persistently infected pregnant rats. AB - Infant rats are susceptible to persistent rat virus (RV) infection, but risk of persistent infection after prenatal exposure to virus is unclear. We examined this aspect of RV infection in the progeny of dams inoculated with virus during or prior to pregnancy. Sprague-Dawley (SD) dams were infected during pregnancy (gestation day 9) by oronasal inoculation with 10(5) TCID50 of the UMass strain of RV. SD rats were infected prior to pregnancy by oronasal inoculation of two day-old females with 10(2) TCID50 of RV-UMass, which induced persistent infection. They were mated to non-immune males after reaching sexual maturity. Rats were assessed for RV infection by virus isolation, in situ hybridization, contact transmission, or serologic testing. The progeny of dams inoculated with virus during gestation had high prevalence of infection through postpartum week 9 (9 of 12 rats were virus positive at week 3, and 7 of 10 were virus positive at week 9). Additionally, 2 of 10 rats were virus positive at least through postpartum week 15. The progeny from persistently infected, seropositive dams had no evidence of infection and did not transmit infection to contact sentinels. However, 12 dams were virus positive at necropsy and 9 had transmitted infection to their breeding partners. These results indicate that prenatal infection in non immune dams can lead to RV persistence in their progeny. By contrast, the progeny of persistently infected dams are protected from infection, presumably by maternal antibody, although their dams can transmit infection to their breeding partners. PMID- 11926301 TI - Analgesic efficacy of orally administered buprenorphine in rats. AB - The analgesic effect of orally administered buprenorphine was compared with that induced by a standard therapeutic injected dose (0.05 mg/kg of body weight, s.c.) in male Long-Evans rats. Analgesia was assessed by measuring pain threshold, using the hot-water tail-flick assay before and after administration of buprenorphine. The results suggest that a commonly used formula for oral buprenorphine in flavored gelatin, at a dose of 0.5 mg/kg, does not increase pain threshold in rats. Instead, oral buprenorphine doses of 5 and 10 mg/kg were necessary to induce significant increases in pain threshold. However, these doses had to be administered by orogastric infusion because the rats would not voluntarily eat flavored gelatin containing this much buprenorphine. The depth of analgesia induced by these infused doses was comparable to that induced by the clinically effective s.c. treatment (0.05 mg/kg). PMID- 11926302 TI - The Charles River "hairless" rat mutation is distinct from the hairless mouse alleles. AB - The Charles River (CR) "hairless" rat is one of the autosomal recessive hypotrichotic animal models actively studied in pharmacologic and dermatologic research. Despite its widespread use, the molecular basis of this monogenic mutation remains unknown, and the skin histologic features of this phenotype have never been described. However, the designation "hairless" has been used as an extension of the hairless mouse (hr) nomenclature on the basis of the clinical absence of hairs in both phenotypes. We present a description of the histopathologic changes in heterozygous and homozygous CR hairless rat mutants during the first month of life. The postnatal homozygous rat skin was characterized by abnormal keratinization of the hair shaft and formation of a thick and dense layer of corneocytes in the lower portion of the epidermal stratum corneum. This layer prevented the improperly keratinized hair shaft from penetrating the skin surface. Starting from the latest stages of hair follicle (HF) development, obvious signs of HF degeneration were observed in homozygous skin. This process was extremely rapid, and by day 12, mainly atrophic HFs with abnormal or broken hairs were present in the skin. Therefore, the mutation in the CR rat abrogates cell proliferation in the hair matrix and affects keratinocyte differentiation in the HF and interfollicular epidermis, a phenotype that is completely distinct from hr/hr. To test whether the CR rat harbored a mutation in the hr gene, we analyzed the coding region of this gene and consensus intron splice site sequences in mutant rats and found no mutation, further supporting phenotypic evidence that the hairless phenotype in CR rats is not allelic with hairless. Finally, using intragenic polymorphisms, we were able to exclude homozygosity at the hairless locus by use of genotypic analysis. Thus, morphologic analysis of successive stages of phenotype development in the CR hairless rat, together with definitive molecular studies, indicate that this mutation may be unique among the other hypotrichotic rat mutations. PMID- 11926303 TI - New nephron development in goldfish (Carassius auratus) kidneys following repeated gentamicin-induced nephrotoxicosis. AB - Renal development in mammalian kidneys can only be studied in embryonic animals. Hence, research in this area is hampered by the need to maintain pregnant animals and by the small size of the embryonic kidney. Here, we describe a goldfish (Carassius auratus) model for studying renal repair and nephron development in an adult animal. Previous studies have indicated that chemically induced nephrotoxicosis in goldfish is followed by new nephron development. We tested the hypothesis that new nephron development is not a one-time only event and, thus, will occur after repeated nephrotoxic events. We used repeated injections of gentamicin (50 mg/kg of body weight), a nephrotoxic antibiotic, which has been used as a model nephrotoxicant to study renal repair. Fish were allowed either a recovery period of 9 or 24 weeks between injections. In both experiments, new nephrons developed after each injection of gentamicin, supporting our hypothesis. Nephron development occurring after a 9-week recovery period was similar to development observed after a 24-week recovery period; therefore, the shorter experimental paradigm appears sufficient and can save time and money. Future research using this fish nephrogenesis model may identify the genes responsible for nephron neogenesis. Such information is a prerequisite for developing alternative renal replacement therapies based on the induction of de novo nephrogenesis in diseased kidneys. PMID- 11926304 TI - Hematologic and blood biochemical variables of captive chimpanzees: cross sectional and longitudinal analyses. AB - Hematologic and blood biochemical variables are of great importance in medical and veterinary practice. In addition, these analytes may have significance as potential biomarkers of aging. Previous reports on normative values of these variables in the chimpanzees are based on cross-sectional studies that did not include individuals of advanced age. To address this omission, we performed cross sectional and longitudinal analyses of hematologic and blood biochemical data collected from chimpanzees over a 9-year period. One-hundred forty-six females and 106 males of ages representing the entire life span of the species were studied. We derived normative cross-sectional values of 14 commonly measured hematologic and 20 blood biochemical variables, which should provide a useful reference for clinical blood studies in chimpanzees. In addition, we found in a cross-sectional regression analysis of our data that most analytes varied significantly between males and females, and that they varied markedly with age. Most variables had year-to-year consistency within the same individuals, as indicated by statistically significant intra-year correlation coefficients. Finally, we performed a longitudinal analysis of the analytes in chimpanzees by calculating the slopes and intercepts of the best-fitting trend line for each individual. The resulting slopes were analyzed by sex and by decade of age of subjects to determine whether trends were consistent. Consistent trends detected in the longitudinal analysis were usually restricted to the first decade of life, and thus represented maturational processes. The overall lack of within-animal trends covering all or most of the period from early adulthood through old age in this 9-year study suggests that a longer period of follow-up than used here may be required to document senescence-related changes. PMID- 11926305 TI - Results of crown-height reduction and partial coronal pulpectomy in rhesus monkeys (Macaca mulatta). AB - BACKGROUND AND PURPOSE: In research facilities using non-human primates, crown height reduction with partial coronal pulpectomy ("vital pulpotomy") is routinely performed on canine teeth of adult male monkeys to reduce self-trauma and the potential for injury to staff or cage-mates. Success of pulpotomy techniques in humans is reportedly 40 to 60%. Failure leads to chronic inflammation and pulp necrosis, which introduces variability in research animals, and may affect research results. The purpose of the study reported here was to determine failure rate of this procedure by evaluating clinical and radiographic findings at 3, 9, and 24 months after crown amputation and partial coronal pulpectomy of maxillary canines in adult male rhesus monkeys. METHODS: Forty-seven maxillary canine teeth from 24 adult male rhesus monkeys were treated by use of crown amputation and partial coronal pulpectomy, using standard dental technique. Follow-up clinical and radiographic examination was performed 3, 9, and 24 months after surgery. RESULTS: At three months after surgery, there was no clinical evidence of failure at any of the teeth. On the basis of radiographic findings, 2 of 47 teeth had failed and one was suspicious for early failure. At nine months, clinical evidence of failure was not apparent; radiographically, 5 of 44 teeth appeared to have failed and 3 others were suspect. Two years post-operatively, failure was clinically evident at two teeth, with radiographic evidence of failure in five teeth, and suspicion of early failure in an additional six of 41 teeth [corrected]. CONCLUSIONS: The failure rate of crown amputation and partial coronal pulpectomy of canine teeth in adult male rhesus monkeys is high, and the chronic inflammation associated with this is cause for concern. PMID- 11926307 TI - Surgical technique for ambulatory management of airsacculitis in a chimpanzee (Pan troglodytes). AB - PURPOSE: Bacterial infections of the air sac have been reported in many nonhuman primates. Approaches to the management of airsacculitis have included combinations of medical and surgical therapies. These strategies have often required repeated attempts to drain exudate from the affected air sac, as well as necessitating that the animal endure isolation or undergo intensive postoperative care before returning to its social group. METHODS: A stoma was created via deliberate apposition of the air sac lining and skin to allow continuous drainage. Antibiotic therapy based on culture and antimicrobial susceptibility of the air sac contents was administered while the chimpanzee remained in its social group. RESULTS: We were able to attain complete resolution of the infection after a course of oral antibiotic therapy. The stoma closed gradually over a three-week period, and the chimpanzee has remained free of infection since that time. CONCLUSION: Despite the severity of the air sac infection in this chimpanzee, we were able to resolve the infection easily, using a simple surgical technique. This method allowed treatment without interfering with social standing or subjection to repeated anesthetic and treatment episodes. This method could be a simple, useful alternative for managing airsacculitis in nonhuman primates. PMID- 11926306 TI - Therapeutic effect of a pig-derived peptide antibiotic on porcine wound infections. AB - PURPOSE: We investigated the therapeutic potential of the pig-derived antimicrobial peptide protegrin-1 (PG-1) against porcine skin wounds infected with Pseudomonas aeruginosa. MATERIALS AND METHODS: Using a porcine skin wound model, PG-1 was added to the wound fluid either at the time of P. aeruginosa inoculation, four hours after inoculation or 24 hours after inoculation. Wound fluids were analyzed 20-24 hours later by use of colony-forming unit (CFU) assays, semiquantitative immunoblot analysis for PG-1, and radial diffusion assays (RDA) for residual in vitro activity. RESULTS: Results of the CFU assays indicated a 10,000-fold decrease in the number of bacteria when PG-1 was added at the time of inoculation, a 120-fold decrease when added 4 hours after inoculation and a 10-fold decrease when added 24 hours after inoculation. Results of immunoblot analysis and RDA indicated that PG-1 concentrations for each of the three conditions remained increased in wound fluid 20 to 24 hours after treatment, and correlated with increased residual in vitro antimicrobial activity. CONCLUSIONS: These results document that the endogenous antibiotic PG-1 significantly prevented the colonization of P. aeruginosa in wounds and reduced the in vivo bacterial concentration in established wound infections. Therapeutics used in the same animal species from which they were derived are a promising means for preventing and treating localized infections. PMID- 11926308 TI - Hip dysplasia in rabbits: association with nest box flooring. AB - PURPOSE: To study etiologic aspects of hip dysplasia in a colony of Dutch-belted rabbits. METHODS: Rabbits used in the study were part of a reproductive toxicologic study. Incidence of hip dysplasia among 296 Dutch-Belted rabbit kits raised on waxed cardboard, smooth Plexiglas, or Plexiglas covered with textured adhesive strips was recorded. All animals were examined at 2 to 4 weeks of age for inability to adduct one or more limbs, then were classified as normal or dysplastic. A subset of 16 juvenile male rabbits (4 normal, 12 affected) raised on Plexiglas flooring were given a physical examination at 12 weeks of age followed by complete necropsy. In four animals (one normal, three affected), pelvic radiography and neurologic examination were performed. RESULTS: Seven percent of the rabbits kits reared on waxed cardboard flooring and 22% of those reared on smooth Plexiglas flooring developed hip dysplasia. Animals reared on Plexiglas floor with traction strips did not have evidence of hip dysplasia. Among the animals selected for detailed analysis, body weight was similar between rabbits with or without splay leg. Affected animals had splaying of one or both hind limbs, various degrees of flattening and reduction of the size of the femoral head, subluxation of the hip, valgus deformity, and patellar luxation. Histologically, there was marked thickening of the hip joint capsule with fibrocartilage formation, mild trabecular bone loss, and bony sclerosis of the proximal portion of the femur and adductor muscle hypoplasia. CONCLUSIONS: Provision of non-slippery flooring during the postnatal period is critical in preventing development of hip dysplasia in rabbits. Hip dysplasia resulted in significant musculoskeletal changes, but not abnormal neurologic development. PMID- 11926309 TI - CPPD crystal-induced suppression of neutrophil apoptosis is regulated by the ERK1/2 and PI3-kinase/Akt pathways. PMID- 11926310 TI - Modulation of inflammation in vivo through induction of the heat shock response, effects on NF-kappaB activation. PMID- 11926311 TI - Characterization of CXC and CC chemokine expression in a murine model of chronic granuloma. PMID- 11926312 TI - Scytonemin--a marine natural product inhibitor of kinases key in hyperproliferative inflammatory diseases. PMID- 11926314 TI - International Society for Rheumatic Therapy guest society symposium: recent therapeutic experiences with new generation drugs for inflammatory diseases. PMID- 11926315 TI - Inflammation and oncogenesis. PMID- 11926313 TI - Atorvastatin activates PPAR-gamma and attenuates the inflammatory response in human monocytes. AB - OBJECTIVE: To investigate the ability of statins to activate the nuclear receptor peroxisome proliferator-activated receptor gamma (PPAR-gamma) in primary human monocytes in culture. MATERIALS AND METHODS: Human peripheral monocytes were incubated with atorvastatin (0.1-10 micromol/1) for up to 24 hours. PPAR-gamma expression was analysed by electrophoretic mobility shift assay. Pro-inflammatory cytokines were measured by enzyme-linked immunosorbent assays, and oxygen consumption was determined polarographically with a Clark-type oxygen electrode. RESULTS: We found that atorvastatin activates PPAR-gamma and inhibits the production of tumour necrosis factor-alpha up to 38% (p < 0.05), monocyte chemoattractant protein-1 up to 85% (p < 0.05), and gelatinase B up to 73% (p < 0.05), in a concentration-dependent manner. Moreover, atorvastatin shows concentration-dependent inhibition of cellular oxygen consumption up to 41%. CONCLUSIONS: These findings contribute to the growing knowledge of the anti inflammatory effects of statins, and have led us to the suggestion that statins may control inflammatory responses by the regulation of intracellular lipid homeostasis. PMID- 11926316 TI - New drugs for the treatment of inflammatory, respiratory and immunological diseases. PMID- 11926317 TI - Signaling and cell death in lymphocytes. AB - After activation, CD4 helper T (Th) cells differentiate into Th1 or Th2 effector cells. These two subsets produce distinct profiles of cytokines and regulate different immune responses. Here we discuss transcription factors and signaling pathways that are selectively expressed or activated in Th1 and Th2 cells to regulate cytokine gene expression, cell proliferation and apoptosis. PMID- 11926318 TI - Functional and biochemical characterization of ADAMs and their predicted role in protein ectodomain shedding. AB - Proteolysis on the cell surface and in the extracellular matrix is essential for normal cellular functions during development and in the adult, but it may also have undesirable consequences by promoting diseases such as cancer, arthritis, and Alzheimer's disease. A particularly interesting function of proteolysis on the cell surface is to release ectodomains of membrane proteins from the plasma membrane. This process, which is referred to as protein ectodomain shedding, affects a variety of proteins with important roles in development and in disease, including cytokines, growth factors, receptors, adhesion proteins and other proteins such as the amyloid precursor protein. In principle, protein ectodomain shedding can dramatically change the properties of a substrate protein. For example, membrane anchored growth factors such as transforming growth factor alpha (TGF-alpha) are only able to activate their receptor, the epidermal growth factor receptor (EGFR), after they are shed from the plasma membrane. Inhibitor studies have implicated zinc-dependent metalloproteases in protein ectodomain shedding, and in particular a family of metalloproteases termed ADAMs (a disintegrin and metalloprotease). The main focus of my lab is to understand the role of different ADAMs in protein ectodomain shedding, and to learn about the functional consequences of protein ectodomain shedding for individual substrates. PMID- 11926319 TI - CD23 shedding: requirements for substrate recognition and inhibition by dipeptide hydroxamic acids. AB - CD23 (low affinity IgE receptor, FcepsilonRII) is expressed as a Type II extracellular protein on a variety of cells such as B cells, monocytes and macrophages and is cleaved from the cell surface to generate several distinct fragments. The expression of CD23 on the cell surface as well as the generation of soluble fragments of CD23 has been shown to be involved in regulation of IgE synthesis. CD23 is released from the cell surface by a metalloprotease, analogous to the cleavage of other cell surface molecules such as TNF-alpha. This activity has been extensively studied with respect to biochemical characterization and ability to cleave specific mutants of CD23. Both local sequence and distal domains have been shown to affect cleavage of CD23. Selective dipeptide hydroxamic acid inhibitors of CD23 processing have been identified and demonstrated to very potently and selectively inhibit CD23 processing. PMID- 11926321 TI - Transtibial energy-storage-and-return prosthetic devices: a review of energy concepts and a proposed nomenclature. AB - Prosthetic devices that can store and return energy during gait enhance the mobility and functionality of lower-limb amputees. The process of selecting and fitting such devices is complicated, partly because of confusing literature on the topic. Gait analysis methods for measuring energy characteristics are often incomplete, leading to inconsistencies in the energy classifications of different products. These inconsistencies are part of the reason for the lack of universally accurate terminology in the field. Inaccurate terminology perpetuates misunderstanding. In this paper, important prosthetic energy concepts and methods for measuring energy characteristics are reviewed. Then a technically accurate nomenclature and a method of functional classification are proposed. This review and proposed classification scheme should help to alleviate confusion and should facilitate enhancement of the design, selection, and fitting of prosthetic limbs for amputee patients. PMID- 11926322 TI - Northwestern University Auditory Test No. 6 in multi-talker babble: a preliminary report. AB - The purpose of this project was to develop a spoken word-recognition task that could be used clinically to evaluate recognition performance of individuals with hearing loss in a background noise. The test instrument incorporated monosyllabic words at seven levels over a 35-dB range presented in a background of "multi talker" babble that was fixed in level. In Experiment 1, we established normative data on 24 young adult listeners with normal hearing and on 50 older adult listeners with high-frequency hearing loss. In Experiment 2, we examined the effects that age and hearing loss have on understanding speech in multi-talker babble by studying 15 subjects in each decade interval from 20 to 79 years. PMID- 11926323 TI - The influence of hospital culture on rehabilitation team functioning in VA hospitals. AB - A conceptual model of rehabilitation effectiveness, in which team functioning is influenced by hospital culture, has been previously suggested by several authors of this study. The current study tested the efficacy of the hospital culture portion of the model using survey data from 523 rehabilitation team members and 162 administrators from 50 participating Veterans Administration Hospitals (VAHs). We assessed four types of hospital culture (personal, dynamic, formal, and production-oriented) using an instrument developed originally for a Competing Values Model. Rehabilitation team members and administrators perceived three of the four hospital cultures differently (p < 0.0001), agreeing only on production oriented culture. With the use of VAH dominant culture as the independent variable, statistically significant differences were found among all nine measures of team functioning (dependent variables). The major contrast was between personal versus formal hospital culture types. PMID- 11926324 TI - Comparison of cycling kinetics during recumbent bicycling in subjects with and without diabetes. AB - We compared recumbent bicycle kinetics in diabetic peripheral neuropathy and nondiabetic men (nine per group). 3D kinematic and force pedal data in a linked segment model were used. The generalized muscle moment (GMM) patterns were similar between the two groups except for (1) decreased maximum knee flexor moment, (2) increased minimum knee flexor GMM, and (3) maximum hip extensor GMM by the diabetic subjects. Similar to the walking support moment, a summation moment immutable pattern was observed, although the groups accomplished it differently. The diabetic group utilized the hip during the power phase and the knee during the recovery phase. The nondiabetic group utilized both joints together during both phases. Differences in ankle GMM were not observed, suggesting further research using the recumbent bicycle as an exercise modality for diabetic peripheral neuropathy patients to enhance ankle range of motion and strength, commonly observed walking deficits. PMID- 11926325 TI - Circuit resistance training in persons with complete paraplegia. AB - BACKGROUND/OBJECTIVE: We assessed the metabolic and heart rate (HR) responses to a single session of circuit resistance training (CRT) in six subjects with complete paraplegia (T5-T12 levels) in order to determine the caloric cost of the exercise. METHODS: Subjects underwent isoinertial weight training exercises with interspersed periods of high-cadence, low-resistance arm ergometry (AE). Following protocol familiarization, subjects completed one session of CRT during which continuous monitoring of HR, oxygen uptake (VO2), and respiratory exchange ratio (RER = VCO2/VO2) was performed. Caloric cost was calculated from the exercise VO2 values across the CRT session. A peak arm exercise test allowed data to be expressed as percentages of peak VO2 and HR. RESULTS: Subjects displayed mean VO2 values of 11.6 +/- 2.4 ml/kg/min (mean +/- SD) and a mean HR of 136 +/- 17 beats/min across the CRT session, corresponding with 49.0% of peak VO2 and 76.8% of peak HR. The RER values ranged from 0.96 to 1.19 and averaged above unity throughout the CRT session. CONCLUSION: Despite the modest absolute VO2 during exercise, CRT satisfies operational criteria developed for cardiorespiratory exercise prescriptions in persons without disability. The RER values recorded indicate that CRT is intense work that relies primarily on glycolytic metabolism. PMID- 11926320 TI - ErbB-4: a receptor tyrosine kinase. AB - ErbB-4 is a receptor tyrosine kinase that is activated by the binding of specific growth factors to its ectodomain. In addition to the initiation of signal transduction pathways that direct cell responses, such as proliferation or differentiation, this receptor is subject to ligand-dependent trafficking events. The signal transduction events are controlled by ligand-dependent activation of the receptor tyrosine kinase activity, which results in receptor autophosphorylation and the tyrosine phosphorylation of other cellular proteins. The trafficking events include migration into and out of membrane microdomains, entry into internalization pathways and endocytosis, plus proteolytic fragmentation. PMID- 11926326 TI - Use of data from nonrandomized trial designs in evidence reports: an application to treatment of pulmonary disease following spinal cord injury. AB - Evidence reports summarize the evidence pertaining to various health-related topics. Including evidence from nonrandomized studies into such reports involves a trade-off between availability and bias. We describe a general framework by which information from nonrandomized studies might be integrated reasonably into evidence reports and illustrate its application to a recent evidence report on preventing pulmonary complications among patients with spinal cord injury. The proposed framework, which is based upon the premise that producing a fair summary of the evidence requires only a level of evidence judged by clinical experts to be sufficient to the task at hand, may help focus scarce resources, strengthen the quality and documentation of decisions including evidence from nonrandomized studies, and suggest high-priority areas for future research. PMID- 11926328 TI - A kinematic study of the upper-limb motion of wheelchair basketball shooting in tetraplegic adults. AB - Kinematic aspects of the reduced shooting ability of tetraplegic (TP) wheelchair basketball players were investigated and compared with those of able-bodied (AB) basketball players. TP showed significantly smaller values for the vertical component of ball release velocity (4.26 (degree/s) versus 5.45 (degree/s)) and maximum wrist flexion angular velocity (878.4 (degree/s) versus 1445.9 degrees) than AB. Moreover, for a specific shoulder horizontal adduction motion, a larger range of shoulder abduction motion and larger displacements of the right shoulder were observed in TP. The reduced ball velocity of TP subjects with lesions at the C7 to C8 levels depended on an insufficient wrist flexion angular velocity, where dysfunction of available musculature may be a causal determinant. Further, the specific motions observed in TP subjects most likely maximize the function of available musculature, thereby partially compensating for the dysfunction of the wrist flexor muscles and contributing to resultant ball release velocity. PMID- 11926327 TI - A database of self-reported secondary medical problems among VA spinal cord injury patients: its role in clinical care and management. AB - An interactive data management (IDM) system for the Spinal Cord Injury (SCI) Service was developed to collect self-reported patient data related to secondary medical complications and to provide feedback to the SCI rehabilitation team. The long-term objective is to improve clinical care through a process of staff review of current rehabilitation programs in the areas of prevalence, prevention, and management. Based on data from the first 99 SCI patients visiting the clinic and hospital after the installation of the IDM system, SCI patients reported high rates of current problems with spasticity (53 percent), pain (44 percent), and pressure ulcers (38 percent). Respiratory (12 percent) and bowel (14 percent) problems were less common current problems. The SCI staff questioned the reportedly high spasticity rates. They thought that the patients' answers might have indicated simply the occurrence of spasticity, rather than the more important issue of severe spasticity that interferes with daily activities. The staff suggested several additional spasticity questions to add to the study. In other areas, only a small percentage of patients wanted to talk with a therapist about prevention of pressure ulcers. Patients who had urinary problems consistently reported five urinary signs (e.g., cloudy urine). The clinical staff found these data informative and stated that they should continue to be collected. PMID- 11926329 TI - Reliability of biomechanical variables during wheelchair ergometry testing. AB - Wheelchair ergometer testing is used to characterize wheelchair propulsion mechanics. The reliability of kinematic and kinetic measures has not been investigated for wheelchair ergometer testing. In this study, test-retest reliability of biomechanical measurements on a wheelchair ergometer was determined during a submaximal endurance test. Ten nondisabled subjects (seven male, three female), inexperienced in wheelchair propulsion, completed three separate submaximal fatigue tests. An instrumented wheelchair ergometer was used to measure handrim kinetics while three-dimensional kinematic data were collected. Analysis of variance was used to determine if measurement differences existed across the tests. Intraclass correlation coefficients (ICC) were calculated to determine the reliability of the measurements. The majority of handrim and temporal variables were found to be reliable. Joint kinematic variables were less reliable, especially those involving wrist movements in the fatigued state. It was concluded that most biomechanical variables obtained during wheelchair ergometry were reliable. PMID- 11926331 TI - Personal characteristics that influence exercise behavior of older adults. AB - Long-term exercise participation among older adults will result in healthier lifestyles and reduced need for health care. A better understanding, therefore, of what influences older individuals to start and maintain exercise plans would be beneficial. The twofold purpose of this study was (1) to create a knowledge base of determinants that influence exercise behavior in older adults and (2) to have health professionals prioritize determinants that affect exercise initiation and adherence in older adults. The expert panel examined nine determinants within the category of personal characteristics: age, gender, ethnicity, occupation, educational level, socioeconomic status, biomedical status, smoking status, and past exercise participation. The experts rated the determinants on importance for influencing exercise behavior of older adults. This expert panel concluded that older adults who are in good health and have a history of exercise activity might be more likely to participate in long-term exercise programs. PMID- 11926330 TI - Preliminary evaluation of wheelchair occupant restraint system usage in motor vehicles. AB - Individuals using wheeled mobility devices (WMDs) often use them as motor vehicle seats during transportation. Wheelchair occupant restraint systems (WORSs), consisting of upper torso and pelvic restraints, are usually mounted to the structure of transit vehicles to secure individuals within their wheelchair seats. This preliminary study attempts to evaluate the use and satisfaction of currently installed vehicle-mounted WORSs for individuals using WMD as seats in motor vehicles. A survey was conducted among 33 adults who use their WMD to travel in motor vehicles. Results from the survey showed that upper torso and pelvic restraints installed in private vehicles are quick, comfortable, and easy to use. However, WORS installed in mass transit and paratransit are often uncomfortable to wear, difficult to reach, and time-consuming to use. This preliminary study documents the growing need for developing alternative WORS that are safe, comfortable, and that allow independent usage for wheelchair occupants while traveling in a motor vehicle. PMID- 11926332 TI - Judging the judges: keeping objectivity in peer review. PMID- 11926333 TI - Photo Quiz. The Diagnosis: pilomatricoma. PMID- 11926334 TI - Botanical briefs: buttercup Ranunculus species L.. PMID- 11926335 TI - A case report of bacillary angiomatosis in a patient infected with human immunodeficiency virus. AB - A man infected with human immunodeficiency virus (HIV) presented with a few-month history of an enlarging friable growth on the medial area of the left foot and a one-week history of bilateral lower extremity edema. Clinical and histologic examination led to a diagnosis of bacillary angiomatosis, and the patient responded to antibiotic therapy We provide an overview of bacillary angiomatosis, a rare disorder that affects immunocompromised patients with CD4 cell counts less than 100/microL. PMID- 11926336 TI - An aggressive treatment for aggressive digital papillary adenocarcinoma. AB - Aggressive digital papillary adenoma (ADPA) and adenocarcinoma (ADPAca) are adnexal tumors that are not often recognized because of their rarity. We present a rare case of ADPAca involving the left middle finger of a 43-year-old man. Histopathological features of ADPAca are distinct from those of other eccrine sweat gland tumors; however, ADPAca may be misdiagnosed particularly for a metastasis of papillary adenocarcinoma originating in the colon, thyroid, or breast. Clinicopathological correlation is essential to ule out a possible risk of metastatic carcinoma of the skin. Recognition of these tumors is important because of a potential risk of local recurrence nd distant metastases. Aggressive surgical treatment consisting of digit amputation is advocated in the treatment of ADPAca. PMID- 11926337 TI - Chilblain lupus erythematosus lesions precipitated by the cold. AB - Lupus erythematosus (LE) may exhibit a broad array of clinical presentations. Cutaneous manifestations include malar rash, discoid lesions, alopecia, and panniculitis. Cold-induced lesions are uncommon. To illustrate this unusual presentation, we describe a case of typical chilblain LE. PMID- 11926338 TI - Eruptive dermatofibromas in a kindred. AB - Benign fibrous histiocytomas of the skin, or dermatofibromas, are common fibrohistiocytic tumors that affect women more commonly than men. Proposed causes range from a reactive process to a neoplastic growth. We report on a family affected with numerous dermatofibromas, suggesting that there also may be a genetic cause to this common tumor. PMID- 11926339 TI - Characteristics of office-based physician visits for cutaneous fungal infections. an analysis of 1990 to 1994 National Ambulatory Medical Care Survey Data. AB - To our knowledge, visits in the ambulatory setting due to cutaneous fungal infections have not been recently characterized. To provide descriptive epidemiology on ambulatory cutaneous fungal infection visits, we analyzed office based physician visits for cutaneous fungal infections recorded in the National Ambulatory Medical Care Survey (NAMCS) from 1990 to 1994. The International Classification of Diseases (ICD-9) was used to define the cutaneous fungal infections. Sampling weights were applied to achieve the nationally representative estimates. From 1990 to 1994, an estimated 21.6 million physician office visits at an estimated cost of $216 million/y (office visit plus medication costs) were made for cutaneous fungal infection diagnoses. The total cost of office visits (without medication costs) was approximately $116 million/y. The total cost of the top 5 medications was approximately $68 million/y. According to an analysis of visits per physician specialist, dermatologists had the largest proportion of visits for cutaneous fungal infections. The cost associated with the diagnosis and management of cutaneous fungal infections is significant. Of all the physician specialists, dermatologists treated the most cutaneous fungal infections. PMID- 11926340 TI - Fixed drug eruption due to doxycycline and metronidazole. AB - Fixed drug eruption (FDE) can be caused by an assortment of drugs. Although cross sensitivity to 2 chemically related drugs has been frequently described, FDE to 2 unrelated agents rarely has been reported. To our knowledge, we report the first such case due to doxycycline and metronidazole. PMID- 11926341 TI - Annular follicular lichen planus. AB - Annular lesions can be found in cases of lichen planus (LP). We present a case with follicular (histopathologic) and annular (clinical) features. To our knowledge, this is the first case of annular follicular LP reported in the literature. PMID- 11926342 TI - Cutaneous malignant fibrous histiocytoma with the characteristics of different variants. AB - We report the case of a 50-year-old woman with cutaneous malignant fibrous histiocytoma (MFH) on the right hypogastric region. A purplish-red blood-filled tumor, approximately 40 mm in diameter, was detected on the region. A histopathologic analysis of the excised tumor showed that it extended from the upper dermis to the subcutaneous tissue over the fascia and, furthermore, that a variety of cells, from highly atypical spindle shaped to histiocytelike, were embedded in the collagenous stroma without forming a capsule. The storiform pattern was not significant. In addition, an area occupied primarily by multinucleated giant cells and rich in vascular components was observed in the deep portion of the tumor that came into contact with the bloody contents. Based on these findings, the patient was diagnosed as having a cutaneous MFH exhibiting the characteristics of different variants. PMID- 11926343 TI - Eosinophilic intranuclear inclusion bodies in a melanocytic nevus. AB - We report a case of eosinophilic or hyaline true intranuclear inclusion bodies in a melanocytic nevus. Although intranuclear pseudoinclusions are frequently found in melanocytic nevi, true intranuclear inclusions are rare. The true intranuclear nature of the inclusions in our case was confirmed with ultrastructural examination. With reverse transcriptase in situ polymerase chain reaction (RT in situ PCR) analysis, eosinophilic bodies stained positive for molluscum-specific primers. This result suggests that such inclusions may be related to molluscum viral infection of melanocytes. PMID- 11926344 TI - Tumid lupus erythematosus. AB - Tumid lupus erythematosus (TLE) is a variant of cutaneous lupus erythematosus. Most patients who present with these skin lesions are young women. The condition clinically resembles polymorphous light eruption, systemic lupus erythematosus (SLE), reticulated erythematous mucinosis, or gyrate erythema. Histopathologically, the lesions resemble classic lupus erythematosus because of their superficial and deep lymphohistiocytic inflammatory infiltrates and dermal mucin. However, unlike classic lupus erythematosus, there is little or no epidermal or dermo-epidermal involvement. Antinuclear antibody test results are usually negative. We describe 4 cases of TLE and discuss the differential diagnosis. PMID- 11926345 TI - The effect of age on the relationship of pulse pressure and left ventricular mass in untreated patients with mild to moderate hypertension. AB - AIM: To study the relationship between left ventricular mass (LMV) and pulse pressure (PP) in mild to moderate hypertensive patients according to age and gender. DESIGN AND METHODS: Two hundred and eleven patients aged 18-65 years, 56% males, with untreated mild to moderate hypertension, were included. A 24-h ambulatory blood pressure (BP) profile was recorded in 204 patients. In 174 of them, an echocardiogram of good quality was obtained. PP was defined as the difference between systolic and diastolic BP. RESULTS: Statistically significant differences were observed in office and ambulatory PP according to the age, with a greater PP among hypertensive subjects younger than 30. Globally, there was a significant correlation between LVM and PP, either with clinical PP (r = 0.17, p = 0.024) or with ambulatory PP (24-h PP: r = 0.18, p = 0.016). When stratifying by age group, the stronger correlations were observed in patients younger than 30. When stratifying by gender, previous results were applicable to men, but, in women, no correlation was observed between PP and LVM in either age group. CONCLUSIONS: an increased PP was observed in hypertensive patients younger than 30 in comparison with middle-aged hypertensive patients. The correlation between LVM and PP was stronger in younger male hypertensive patients, but not in female. PMID- 11926346 TI - Nocturnal blood pressure predicts left ventricular mass index in normotensive elderly. AB - Blood pressure (BP) predictors of left ventricular mass index (LVMI) were studied in 40 healthy normotensive (71.4 +/- 4.4 years) and 31 hypertensive (73.5 +/- 4.8 years) elderly community-dwelling subjects using short-axis cardiac cine magnetic resonance imaging and 24-h ambulatory BP monitoring. Mean night-time BPs were calculated from the average of readings during sleep and mean daytime BPs were calculated from the remaining recordings. The hypertensive subjects were all receiving anti-hypertensive therapy with angiotensin-converting enzyme (ACE) inhibitors, calcium-channel blockers, beta-blockers or diuretics. Nocturnal systolic BP was a strong predictor of LVMI in both normotensive (beta = 0.38, p = 0.02) and treated hypertensive (beta = 0.39, p = 0.03) subjects. By contrast, daytime systolic BP was a weaker predictor of LVMI in the treated hypertensives (beta = 0.36, p = 0.04) and did not predict LVMI in the normal subjects (beta = 0.27, NS). Nocturnal BP may partly explain the increase in LVMI with ageing in subjects thought to be normotensive on the basis of daytime clinic BP recordings. PMID- 11926347 TI - The effects of mental stress and the cold pressure test on flow-mediated vasodilation. AB - As a stressful lifestyle has been associated with coronary heart disease, this study aim to evaluate how two stressful tests influence endothelial-dependent vasodilation evaluated by flow-mediated dilation (FMD) of the brachial artery. FMD following 5 min of occlusion of the forearm was evaluated in young healthy volunteers with measurements of brachial artery diameter and blood flow (BABF) using ultrasound before and during a mental arithmetic task (MAT), and during cold pressure test (COP). MAT and COP increased blood pressure to a similar degree (18-21 mmHg). The COP induced a significant reduction in FMD compared to the baseline levels (from 9.3 +/- 3.9 to 5.9 +/- 3.2%, p < 0.01), as well as to the MAT, which in itself did not influence FMD (10.1 +/- 5.0%). However, as MAT increased BABF during hyperaemia significantly, the FMD to BABF ratio was significantly reduced by both stress tests (p < 0.05). Time-control experiments showed FMD measurements to be reproducible and that MAT and COP by themselves only marginally affected brachial artery diameter. In conclusion, cold pressure stress induced an impairment in FMD, but no effect was induced by a MAT. However, when FMD was normalized for the degree of hyperaemic blood flow, the driver of the vasodilation in the brachial artery, a reduction in flow-mediated vasodilation was seen during both mental and cold pressure stress. PMID- 11926348 TI - Insulin resistance and lipids in hypertensive women on hormone replacement therapy. AB - Hypertension, hyperinsulinaemia and dyslipidaemia are strong and independent risk factors for cardiovascular diseases. Their increasing frequency in postmenopausal women suggests that oestrogen deficiency may be a contributing factor. It is well known that oestrogen replacement therapy in postmenopausal women improves carbohydrate and lipid metabolism, but the effect of combined hormone replacement therapy (HRT) remains unclear. The purpose of the present study was to evaluate the effect of HRT on blood pressure, carbohydrate metabolism and lipid profile in postmenopausal women with primary arterial hypertension. The study population consisted of 76 postmenopausal women (mean age 51.1 +/- 6.8 years). Forty hypertensive women received HRT (17-beta-oestradiol, norethisterone acetate, TTS, Estracomb Novartis), whereas 36 women remained without hormonal therapy. One-year combined transdermal HRT did not affect significantly blood pressure and blood pressure variability. HRT was shown to improve lipid profile with a significant decrease in total cholesterol as early as at 3 months. It does not influence carbohydrate metabolism parameters studied by glycaemia and insulinaemia in a standard oral glucose tolerance test. In conclusion, combined percutaneous HRT may reduce the lipid-depended cardiovascular risk in postmenopausal women with arterial hypertension. PMID- 11926349 TI - Excess morbidity and cost of failure to achieve targets for blood pressure control in Europe. AB - AIM: To estimate the burden of failing to achieve targets for blood pressure (BP) control in France, Germany, Italy, Sweden and the UK. METHODS: A cost of illness model was constructed to estimate the impact of uncontrolled hypertension to each national healthcare system. Prevalence of uncontrolled hypertension was taken from published data. Relationships between achieved BP and the cardiovascular events of symptomatic acute myocardial infarction, congestive heart failure and stroke were estimated from the HOT study. Costs were taken from public sources. The acute medical costs of these events were estimated at current prevalence of uncontrolled hypertension and if BP were treated to target. RESULTS: The model estimated that 29 million adults in the five countries (13% population) have BP levels above 160/95 mmHg, and an additional 46 million (21% population) have BP in the range 140/90-160/95 mmHg. The model estimated that healthcare system costs of 1.26 billion euros could be avoided if hypertension management did achieve BP targets. This does not consider the cost of interventions required to reduce the risk of cardiovascular disease. CONCLUSIONS: Failing to achieve BP targets contributes substantially to healthcare system costs and preventable events in the countries studies. PMID- 11926350 TI - The price you pay for hypertension management: incorporation of economic factors into clinical decision making. PMID- 11926351 TI - The potential economic consequences of cognitive improvement with losartan. AB - In a clinical trial, treatment of mild-moderate hypertensive patients with losartan (50 mg) increased Mini-Mental State Examination (MMSE) scores by 4 points from baseline over a 26-month period, compared with a 1-point increase in patients treated with hydrochlorothiazide (25 mg). This study explores the potential economic consequences of this improvement in cognitive function in a population of elderly hypertensive patients in Sweden. Resource use and MMSE data for 437 hypertensive, non-demented subjects aged 75 years and above, were taken from a population-based study in Sweden. MMSE scores were strongly related with costs of care due to higher utilization of home help and special living arrangements in patients with low scores. A 1-point difference in MMSE was associated with a difference in the annual cost of care of approximately 5700 Swedish kronor (SEK). Over 26 months, the potential cost savings from the 4-point improvement observed with losartan was estimated to be between 24700 and 43700 SEK. This can be compared with the acquisition cost of losartan; approximately 5700 SEK over the study period. Thus, an improvement in cognitive function of the magnitude documented in the study of losartan vs hydrochlorothiazide, may translate into economic benefits beyond those expected in terms of blood pressure control. PMID- 11926352 TI - The efficacy of valsartan in essential hypertension and its effects on left ventricular hypertrophy. AB - Recent studies demonstrate that, treatment with angiotensin-converting enzyme (ACE) inhibitors and angiotensin-II receptor blockers may decrease left ventricular hypertrophy (LVH) in hypertensive patients. We wanted to assess the efficacy of valsartan on echocardiographic variables of LVH in 30 mild to moderate hypertensive patients. Valsartan was found effective in controlling hypertension, also echocardiographic indices of LVH such as interventricular septum and posterior wall thickness were reduced and left ventricular mass decreased significantly. Thus valsartan favorably influences cardiac structure in hypertensive patients. PMID- 11926354 TI - Powder sampling. AB - The factors involved when sampling powder mixes have been reviewed. The various methods are evaluated (manual, automatic, and sub-sampling) and the errors incurred are discussed. Certain rules have been applied to various samplers and their suitability for powder mixtures are described. The spinning riffler is apparently the most suitable, while the use of sample thieves should be avoided due to error and bias. PMID- 11926353 TI - The alpha1-Na/K pump does not mediate the involvement of ouabain in the development of hypertension in rats. AB - The Na/K pump of vascular smooth muscle cells (VSMC) and renal epithelial cells (REC) is viewed as a target of digitalis and endogenous ouabain (EO), leading to the development of hypertension. In this study, we compared the effect of ouabain on Na/K pump activity and the intracellular content of monovalent cations in VSMC and REC obtained from rats, humans and dogs. In VSMC from the rat aorta, ouabain inhibited maximal Na/K pump activity measured as the rate of 86Rb influx in Na+ loaded cells, with an ID50 of approximately 20-30 microM without any differences between two strains of normotensive rats (WKY and BN.1x) and three substrains of spontaneously hypertensive rats (SHR). Half-maximal inhibition of the Na/K pump in REC from the rat inner medullary collecting duct was observed at approximately 20 microM of ouabain. In contrast to rat cells, half-maximal inhibition of 86Rb influx in VSMC from human coronary arteries and in REC from the Madin-Darby canine kidney was seen at approximately 0.03 and 0.1 microM ouabain, respectively. At concentrations lower than 100 microM, ouabain did not affect the intracellular content of exchangeable Na+ and K+ in rat VSMC, measured as the steady-state distribution of 22Na and 86Rb, whereas in human VSMC, it increased the intracellular Na+/K+ ratio with an ID50 of approximately 0.5 microM. Keeping in mind that the circulating level of administered digitalis and EO does not exceed 10(-9) M, our results strongly suggest that the involvement of these compounds in the pathogenesis of hypertension in rats is not mediated by inhibition of the alpha1-isoform of the Na/K pump in VSMC and REC. Alternative mechanisms of the involvement of EO and ouabain-like factors in the development of hypertension are considered. PMID- 11926355 TI - Influence of manufacturing process on tabletting ability of powder: comparison between blending, grinding, and spray drying of two formulations made of theophylline and lactose/cellulose or Cellactose. AB - Three manufacturing processes were applied to two formulations composed of 20% anhydrous theophylline associated with either 20% microcrystalline cellulose and 60% lactose or 80% Cellactose. The processing method (dry blending, grinding, or spray drying) and the formulation were investigated through the comparison of the physical and flow characteristics and the compactibility of the end products. The results demonstrated that the formulation had a major effect on the mechanical properties, with binary blends exhibiting a higher resistance than ternary ones, whereas flow properties and densification depended on the process. Nevertheless, it was also observed that spray drying decreased the difference between the mechanical properties of the two formulations, probably by modifying the texture of the Cellactose in suspension. PMID- 11926356 TI - Controlled-release naproxen using micronized ethyl cellulose by wet-granulation and solid-dispersion method. AB - This study has been undertaken to develop a controlled-release tablet dosage form of naproxen using ethocel (ethyl cellulose) as the rate-controlling polymer. The formulations were made by employing the conventional wet-granulation method and the solid-dispersion method. Tablets made by both methods were compared for their controlled-release dissolution profiles. Both methods were useful in developing the controlled-release formulations of naproxen with desirable properties. However, the amount of polymer required to make a formulation with the desired release profile was 33% less via solid dispersion than via wet granulation. A cumulative 88% of naproxen was released from the solid-dispersion formulation, compared with 84% from the wet-granulation formulation. PMID- 11926357 TI - Preformulation studies for an ultrashort-acting neuromuscular blocking agent GW280430A. I. Buffer and cosolvent effects on the solution stability. AB - GW280430A is an ultrashort-acting neuromuscular blocking agent targeted at muscle relaxation to facilitate surgical intubation. The objective of this work was to study the buffer and cosolvent effects on the solution stability of GW280430A. The buffer catalytic effect was examined in citrate, malate, tartrate, and glycine by measuring the rate of degradation of GW280430A (0.2 mg/mL) at constant pH (3), ionic strength (0.15 M), and various buffer concentrations (0.01-0.05 M). The temperature dependence of the buffer catalytic effect and the degradation of the GW280430A in cosolvent (ethanol, propylene glycol, polyethylene glycol 400, N,N-dimethylacetamide)/water mixtures were studied at 40, 50, and 60 degrees C. The loss of parent drug was monitored by reverse-phase high-performance liquid chromatography. The degradation of GW280430A followed first-order kinetics in all buffer solutions. Significant buffer-catalyzed hydrolysis of GW280430A was observed with citrate, tartrate, and malate buffers, but not in glycine-buffered solutions. The activation energies in all buffered drug solutions ranged from 70 to 80 kJ/mol and decreased with increasing buffer concentration. GW280430A degradation was primarily through ester hydrolysis and followed first-order kinetics in aqueous solutions. In cosolvent/water mixtures, new degradation products were observed, indicating a chemical reaction between GW280430A and cosolvents. The reaction activation energies in the cosolvent/water mixtures ranged from 75 to 85 kJ/mol, with the longest t(0.9) at 5 degrees C equal to approximately 12 months and at 25 degrees C equal to 36 days. Consideration should be given to the incorporation of glycine or a low concentration of citrate, malate, or tartrate buffer in the parenteral formulation development of GW280430A. Cosolvents prolonged the predicted t(0.9) for GW280430A in solution, but the enhancement was not significant enough to pursue a liquidformulation. PMID- 11926358 TI - A degradation study of a series of chloromethyl and diazomethyl ketone anti leukemic agents. AB - The chemical stability of a novel cysteine chloromethyl ketone derivative (HI 131) with anti-leukemic activity has been investigated in a microemulsion formulation. HI-131 degrades to two major products, most likely by undergoing oxidation and further reaction with another HI-131 molecule to form higher molecular weight oligomers of the original compound. The degradation kinetics of HI-131 have been studied as a function of pH, buffer composition, ionic strength, and temperature. Degradation follows pseudo-first-order kinetics and the temperature effect obeys the Arrhenius equation. The pH-rate profile demonstrates HI-131 is most stable at lower pH values, although there is no significant influence of ionic strength and buffer ions on the degradation rate. The chemical stability of a homologous series of chloromethyl and diazomethyl ketone derivatives of HI-131 has also been investigated in microemulsion. The relationship between the chain length of the derivatives and the stability is presented. Changing the chloro group to a bromo group resulted in an increase in degradation rate. Alterations to the group on the nitrogen were also investigated. The changes to the stability are discussed in terms of their mechanistic implications. PMID- 11926359 TI - Characterization of film-coated aerosol canisters using electrochemical impedance spectroscopy. AB - Drug adhesion to the walls of an aerosol canister can be prevented/reduced by coating the canister with a hydrophobic polymer (e.g., a fluoropolymer). In this study, three batches of fluoropolymer-coated canisters were investigated by electrochemical impedance spectroscopy (EIS) and scanning electron microscopy (SEM). The EIS technique showed that only one of the batches presented as a monolithic, non-porous film. The other two batches were either partially porous or highly porous. Scanning electron micrographs showed evidence of cracks, within the films, but could not alone establish the porous nature of these defects. For the non-porous and partly porous films it was possible to use the EIS data to determine the approximate film thickness. Estimates of 2-4 microm were obtained for the mean film thickness. These values compared favorably with micrometer estimates obtained following acid dissolution of the aluminum canister. It remains to be seen whether the properties of the films (i.e., the porosity and film thickness, determined by EIS) translate to differences in drug adhesion. Nevertheless, the EIS technique was shown to be a powerful, non-destructive method that lends itself to the rapid analysis of batch-to-batch variation in film-coated canisters. PMID- 11926360 TI - Polysaccharides as excipients for colon-specific coatings. Permeability and swelling properties of casted films. AB - Oligosaccharides such as inulin (In) and polysaccharides such as galactomannans, combined with polymethacrylates on isolated films for film coatings, were obtained from aqueous-based solvents and investigated as potential vehicles for colonic drug delivery. These compositions, which are susceptible to fermentation by colonic microflora, constitute promising excipients for the development of new colon-specific therapeutic systems. The characteristics of several compositions have been demonstrated in permeability and swelling studies on isolated films composed of a polymethacrylate associated with In or galactomannans of mesquite seed gum (MSG). Results reported prove a dependency of the properties of mixed films on the polymethacrylate-polysaccharide concentration ratio and on the composition of the dissolution media. An increase in permeability through the mixed films was observed in a simulated colonic environment for the following compositions: Eudragit RS30D-MSG 70:30 w/w; Eudragit RS30D-In 90:10 w/w; Eudragit RS30D-In 76:24 w/w. PMID- 11926361 TI - Acetylation enhances the tabletting properties of starch. AB - The aim of this study was the evaluation of starch acetate (SA) powders used as tablet excipients. Deformation during powder volume reduction, strain-rate sensitivity, intrinsic elasticity of the materials, and tensile strength of the tablets were examined. Results showed that SA with the lowest degree of substitution (ds) still possessed characteristics of native starch granules. Due to dissolution in synthesis, the properties of higher ds SAs depended on precipitation and drying processes. The acetate moiety, perhaps in combination with existing hydroxyl groups, was a very effective bond-forming substituent. The formation of strong molecular bonds increased, leading to a very firm and intact tablet structure. Small changes existed in compression-induced deformation due to acetylation. Some fragmentation was induced due to the slightly harder and more irregular shape of high-substituted SA particles. The plastic flow under compression was enhanced. Acetylated material was slightly less sensitive to fast elastic recovery in-die, but somewhat more elastic out-of-die. In spite of their superior bonding, SAs under compression behaved similarly to native starches. It was concluded that deformation properties were more the consequence of the molecular chain structure properties of the starch polymer than the effect of the acetate moiety itself. In contrast, the opposite seemed to be the case with the extensive improvement in bond-forming properties. PMID- 11926362 TI - A high-throughput HERG potassium channel function assay: an old assay with a new look. AB - In this paper, we describe an assay using radioactive rubidium (86Rb) efflux to screen functional human ether-a go-go-related gene (HERG) K+ channels in a high throughput screening (HTS) format. This assay offers an alternative way to examine junctional interactions between chemical compounds and HERG K+ channels. Follow-up experiments and discussions were carried out to address a variety of factors that affect potency evaluation within the Rb efflux assay. Factors that can affect the assay results, such as assay time, efflux rate, and compound blocking kinetics, are discussed in detail. Our results provide some explanations for the variances of the assay results and offer some guidelines for using the Rb efflux assay to evaluate compound interactions with HERG K+ channels in the pharmaceutical industry. PMID- 11926363 TI - Profiling of drugs for membrane activity using liposomes as an in vitro model system. AB - The increasing size of chemical libraries being analyzed by high-throughput screening results in a growing number of active compounds that need to be assessed before moving forward in the drug development process. As a consequence, more rapid and highly sensitive strategies are required to accelerate the process of drug discovery without increasing the cost. Due to the fact that significant numbers of compounds from combinatorial libraries are hydrophobic in nature, approaches are needed to evaluate the potentialfor these compounds to interfere with the functions of biological membranes. The liposome system was used to detect agents that act as follows: (i) ionophores able to induce specific ion permeability, e.g., valinomycin for K+ and protonophoric uncouplers for H+; (ii) ion antiporters which exchange H+ for other ions, e.g., nigericin; (iii) agents that form low specificity ion channels in the membrane, e.g., gramicidin; and (iv) detergents and other membrane-disrupting agents. We propose using this liposome assay during the drug development process to identify compounds that have membrane activity and, as a consequence, produce a biological effect by altering the physico-chemical properties of the cell membrane rather than interacting with a protein target. Screening of a representative set of biologically-active compounds (198) indicated that the majority of systemic antimicrobial drugs, but not topical drugs, lack membrane activity in this model system. PMID- 11926364 TI - Solutol and cremophor products as new additives in suppository formulation. AB - Our research has a double purpose. On the one hand, doctors have expressed the need to formulate a rectal suppository dosage form from diuretic ethacrynic acid, which would add to the choice of treatment methods and thereby increase the possibilities of individual cure. On the other hand, the liberation and thereby the bioavailability of poorly-soluble ethacrynic acid needs to be enhanced, and for this purpose solubility-increasing additives new to rectal therapy were used. Solutol HS 15, Cremophor RH 40, and Cremophor RH 60 were used as additives in concentrations of 1, 3, 5, and 10%. The quantity of drug released changed as a function of additive concentration. Depending on the acceptor phase, the best results were achieved with an additive concentration of 1-3%, which is related to the optimal additive quantity accumulated on the boundary surface. PMID- 11926365 TI - Investigation of potential ionic interactions between anionic and cationic polymethacrylates of multiple coatings of novel colonic delivery system. AB - The objective of this work was to investigate potential interactions between anionic (Eudragit FS) and cationic (Eudragit RL) polymethacrylates of multiple coatings of a novel colonic drug delivery system. Aqueous films of pure polymers Eudragit FS (FS) and Eudragit RL (RL) and their superimposedfilm (FS-RL) were cast on glass slabs. The potential ionic interactions were studied by analysing the dried films using differential scanning calorimetry (DSC), Fourier transform infrared spectroscopy (FT-IR), and nuclear magnetic resonance (NMR). The glass transition temperatures (Tg) of pure RL and FS were 60 degrees C and 22 degrees C, respectively; FS-RL showed two distinct glass transitions at 59 degrees C and 24 degrees C in the second heating cycle. In the 13C-MAS spectra of the samples in the solid state, no shifts of the resonance could be detected in the superimposed film compared with the pure polymers. The FT-IR spectra of the superimposed film did not show any significant shift of the bands of the -NMe3+ group of RL and the -COO- function of FS compared with the spectra of the pure polymers. No ionic interactions between anionic and cationic polymethacrylates were revealed by DSC, FT-IR, and NMR. PMID- 11926366 TI - Study of the influence of the pH media dissolution, degree of polymerization, and degree of swelling of the polymers on the mechanism of release of diltiazem from matrices based on mixtures of chitosan/alginate. AB - The dissolution profiles of formulations based on mixtures of chitosan/alginate depend on the pH. It is possible to distinguish two processes: (a) a fast kinetic drug release up to 180 min, where the pH value changes from 1.17 to 2.21 and the drug released is controlled by the degree of polymerization and the quantity of chitosan in the formulation; (b) a low kinetic drug release between 210 and 480 min, where the pH value changes from 5.52 to 8.72 and the drug release from the matrix is controlled by the interpolymeric complex. In all formulations the order of release, according to Peppas's model in the range of fast kinetic drug release, was between 0.5 and 1.0. The mechanism of release was non-fickian diffusion, which corresponds to a coupling mechanism of diffusion and relaxation of the polymer. PMID- 11926367 TI - Response components of LiF:Mg,Ti around a moderated Am-Be neutron source. AB - The responses of TLD-1010, TLD-700 and TLD-600 thermoluminescence dosemeters to the radiation field inside a water tank enclosing an isotopic 241Am-Be neutron source are analysed. Separate contributions coming from thermal neutrons, neutrons with energies above thermal and gamma rays to the total response of the three types of TLD are obtained. This is accomplished by assuming that the gamma responses for materials with different 6Li enrichments are identical and that the neutron response of TLD-700 is negligible compared to TLD-100 and TLD-600. The last assumption is tested by Monte Carlo simulations of the neutron energy spectrum at the points where the TLDs are irradiated. PMID- 11926368 TI - Intercomparison of whole-body counters by using a subject who had incorporated 137Cs into the body. AB - During the years 1996-2000, eight whole-body counting facilities (WBC) from Finland, Germany, Japan and Russia took part in an intercomparison using a resident of the Russian town of Novozybkov who had been seriously contaminated as a result of the Chernobyl accident. The subject R (adult male, height 172 cm average body mass 64 kg; and 137Cs body burden within the range of 1-15 kBq) was investigated in the participating institutions during his business trips. The experimentally obtained data for his 137Cs body burden were compared with the predicted values, which had been deduced from the measurements of subject R using the reference WBC (St Petersburg Institute of Radiation Hygiene) and from his effective half-time of 137Cs in the body (68 days). The obtained results did not deviate more than 20% from reference activities. Four facilities were able to quantity the 40K in the subject's body. The differences between reported values of potassium did not exceed 10%. For subject R, the average annual effective dose from radiocaesium was 0.25 mSv and it was 0.18 mSv from 40K in the years 1996/97. The reliability of using a subject with naturally incorporated artificial radionuclides ('walking standard') instead of an anthropomorphous phantom for calibration and intercomparison of whole-body counters in a large-scale nuclear accident is discussed. PMID- 11926369 TI - Bayesian internal dosimetry calculations using Markov Chain Monte Carlo. AB - A new numerical method for solving the inverse problem of internal dosimetry is described. The new method uses Markov Chain Monte Carlo and the Metropolis algorithm. Multiple intake amounts, biokinetic types, and times of intake are determined from bioassay data by integrating over the Bayesian posterior distribution. The method appears definitive, but its application requires a large amount of computing time. PMID- 11926370 TI - Modelling the 137Cs ingestion dose from consumption of marine fish in Hong Kong. AB - This paper presents a compartmental model for estimating the ingestion dose, due to 137Cs, arising from the consumption of marine fish in Hong Kong. 137Cs is one of the more important radionuclides released in routine liquid effluents discharged from the Guangdong Nuclear Power Station at Daya Bay, which began commercial operation in 1994. In the model, three sea/ocean compartments are considered. Assuming the discharge of this radionuclide is maintained at a constant rate, the model shows that the concentration of 137Cs in the water and in the marine fish in the three sea compartments would become steady after 5 years. The predicted annual dose to an average local individual in Hong Kong, for a release rate of 10 GBq.y(-1), is 3.2 x 10(-5) microSv, which is dominated by the contribution from fish cultured in Hong Kong waters. The cumulative collective dose to the local population of 6 million, at 50 years of discharge, amounts to 9.0 x 10(-3) man.Sv. The annual dose to members of the critical group of local fish farmers does not exceed 3.0 x 10(-3) microSv. All these doses are small compared to the dose of around 1.2 microSv.y(-1) arising from ingestion of naturally occurring radionuclides found in marine fish. Sensitivity of model parameters and uncertainties of prediction are also studied. Difficulties encountered in model validation are discussed. Despite such difficulties. limited field data that are available show that the predicted results are generally within one order of magnitude with measurements. PMID- 11926371 TI - Use of a model data set to test methods for mapping radon potential. AB - It is difficult to test the accuracy of mapping methods using real measurement data of radon levels in houses, as different methods produce different maps, and there is no standard to compare them with. A model data set, Linevar, was therefore constructed, having features which allow the accuracies of different mapping methods to be compared. Data for Linevar were generated such that they had a geometric mean that varied across the map in a pattern of stripes of higher and lower radon potential. with widths of 1, 2, 4, and 8 km over an area 100 km square. The geometric standard deviation was kept at a constant value. Data were generated at densities varying from 1 to 100 results per 1 km grid square in different parts of the map. The data set was used to develop a mapping method that shows as much detail as possible without generatin, artefacts. PMID- 11926372 TI - Intercomparison exercise of the PTB, BfS, MPA and PSI calibration facilities for radon gas concentration. AB - The traceability chain of one national reference laboratory (PTB) and three accredited radon calibration laboratories (BfS MPA and PSI) to internationally acknowledged radon gas standards is specified. As an additional tool for quality assurance, interchange of an electronic radon measuring instrument was used as a means for a relative comparison of the radon gas reference atmospheres. The instrument was exposed to radon gas activity concentrations between 500 Bq.m(-3) and 15 kBq.m(-3). Measured sensitivities of the participants agree well inside the range of specified calibration uncertainties. PMID- 11926373 TI - Exposure to 222Rn in ten underground mines in Iran. AB - Concentrations of 222Rn were measured in ten non-uranium underground mines in Iran, including three metal mines and seven coal mines. Two active techniques were applied for these measurements (1) activated charcoal and (2) scintillation cell. A maximum mean concentration of 1.33 kBq.m(-3) was found in the Robat-Karim manganese mine with a hot spot of 12 kBq.M(-3), while the concentration in the Venarge-Qom manganese mine was less than 10 Bq.m(-3), leading to annual effective dose equivalents of 8.3 mSv and (0.06 mSv respectively. The mean concentration levels of 222Rn in coal mines were found to be almost 500 Bq.m(-3), leading to annual effective dose equivalent of 3 mSv. PMID- 11926375 TI - Radon dynamics and reduction in an underground mine in Brazil. Implications for workers' exposure. AB - This work was aimed at studying the behaviour of 222Rn in an experimental underground copper mine in Brazil with a single entrance. The 222Rn concentrations, meaured by using a dynamic radon measuring technique. varied between 30.5 Bq.m(-3), during ventilated conditions applied to the mine galleries, and 19.4 x 10(3) Bq.(-3) for non-ventilated conditions and when operational mining activities were conducted inside. High radon concentration surges were observed after blasting and drilling activities. In the cases of inadequate ventilation, it was estimated that workers could be subjected to exposures as high as 10 microSv.h(-1), only due to 222Rn and its short-lived progeny. The results show the importance of real-time measurements to evaluate radon dynamics during mining operations. PMID- 11926374 TI - Study of natural radionuclide concentrations in an area of elevated radiation background in the northern districts of Bangladesh. AB - The activity concentrations of naturally occurring radioactive materials in soil samples from an elevated radiation background area of three northern districts of Bangladesh were determined using gamma ray spectrometry. The outdoor and indoor external effective dose rates and the radiation hazard indices from these soil activities were evaluated. The dose rates were found to be about four times higher than the reported world average value. The concentration of natural radionuclides, derived radium equivalent activities and the representative level indices were also found to be higher. Recommendations on radiological and dosimetric measures have been suggested with an aim of minimising the harmful effects of ionising radiation to the population of the area concerned. PMID- 11926376 TI - Is it useful to assess annual effective doses that are less than 100 mSv? AB - Questions such as 'How small is small?' and 'How low is low enough?' have long plagued radiation dosimetrists and risk management personnel. Unfortunately, our knowledge about the biological effects of low levels of ionising radiation is scarce and uncertain. If we look to the results of epidemiological studies, we find that it is not easy to arrive at firm conclusions. However, some current radiobiological experiments using microbeams of various radiations, along with improved theoretical models of radiation action, may shed new light on the effects of low levels of ionising radiation. What shall we do in the meantime? Both of our debaters agree that monitoring of radiation workers is necessary, yet careful consideration must be given to the rationale for providing personal monitoring. There is no question that we have done a good job of protecting radiation workers for many years, but we also must be aware of the many implications of our efforts. PMID- 11926377 TI - [18F]FLT; portrait of a proliferation marker. PMID- 11926378 TI - Diagnosis, characterisation and evaluation of treatment response of frostbite using pertechnetate scintigraphy: a prospective study. AB - No large prospective study is available on use of a radionuclide method to assess frostbite and its response to conservative treatment. Recently, a large retrospective study using two-phase bone scintigraphy suggested that non-uptake (or low uptake) in frostbite lesions in the blood pool or bone phase had a strong correlation with the subsequent need for amputation. In the present study, we performed early blood pool scintigraphy with technetium-99m pertechnetate in 20 patients (10-180 days after frostbite injury; 71 clinical lesions) referred for a topical treatment protocol for mild to moderately severe frostbite, and in 22 asymptomatic controls. The aims of the study were to define the "normal" microcirculation of the hands and feet, to characterise the appearances of frostbite lesions on scintigraphy and correlate these with the clinical presentation, and to evaluate the usefulness of scintigraphy for assessing the treatment response. Six patients were "fresh", 12 had been undergoing conservative treatment (topical homeopathic) for several weeks and two had permanent fibrotic hand phalanges. Nine patients on treatment underwent serial scintigraphy a total of 22 times. Eleven patients were imaged only once, including the two with permanent fixed defects. Three distinct physiological microcirculation patterns were evident in control hands or feet: right- or left handedness (seven subjects), subclinical inflammatory foci correlating with callosity or pressure points (36 lesions) and distinctly increased pooling of radiotracer at the thenar or hypothenar region (eight subjects). In frostbite patients, 128 lesions (48 inflammatory, 80 ischaemic) were seen on the scan. This number included all 71 lesions identified clinically, the rest being subclinical. Subclinical frostbite lesion as an entity has not been described previously. Microvasculature of normal muscle groups (in the hands, elbow or feet) was shown on scintigraphy to improve after topical treatment. The treatment resulted in improved or normalised perfusion scan in 25 of 45 ischaemic segments with matching clinical improvement, and reduction or normalisation of uptake in inflammatory lesions (n=30). The data confirm the vasodilator and anti inflammatory action of the topical preparation, and the sensitivity of the radionuclide method in monitoring treatment response. In contrast to the previous findings that non-uptake on a scan performed 2 or more weeks after exposure correlates with the need for amputation, none of the 80 ischaemic lesions in our series required amputation while 60% showed improvement or normalisation in microvascular blood flow even though they were 10-180 days old. Our findings suggest that moderate to severe frostbite lesion can be classified as having infarcted, ischaemic or hibernating (viable) tissue, similar to the classification employed when using myocardial scintigraphy. Absence of uptake even after the initial 10 days does not necessarily indicate infarction and the need for amputation as many such lesions retain potential for vasodilation and recovery. PMID- 11926379 TI - Usefulness of 11C-methionine PET in the evaluation of brain lesions that are hypo or isometabolic on 18F-FDG PET. AB - The fact that some brain tumors show hypo- or isometabolism on fluorine-18 fluorodeoxyglucose positron emission tomography (FDG PET) has caused problems in the detection of primary or recurrent tumors and in the differentiation from benign lesions. We investigated the usefulness of carbon-11 methionine PET in characterizing brain lesions under these conditions. 11C-methionine PET was performed in 45 patients with brain lesions (in 34 for initial diagnosis and in 11 for detection of recurrence) that showed hypo- or isometabolism compared with normal brain tissue on FDG PET. Ten minutes after the injection of 555-740 MBq of 11C-methionine, attenuation-corrected brain images were obtained with a dedicated PET scanner. The brain lesions comprised 24 gliomas, five metastatic brain tumors, four meningiomas, two other brain tumors and ten benign lesions (including three cases of cysticercosis, two cases of radiation necrosis, one tuberculous granuloma, one hemangioma, one benign cyst, and one organizing infarction). Proliferative activity was measured using the Ki-67 immunostaining method in glioma tissues. Thirty-one of 35 brain tumors (89% sensitivity) showed increased 11C-methionine uptake despite iso- or hypometabolism on FDG PET. By contrast, all ten benign lesions showed decreased or normal 11C-methionine uptake (100% specificity). Twenty-two of 24 gliomas (92%) showed increased 11C methionine uptake, the extent and degree of which exceeded 18F-FDG uptake, and the 11C-methionine uptake correlated with the proliferation index (r=0.67). The mean (+/-SD) uptake ratios of glioma to normal brain on FDG and 11C-methionine PET were 0.92+/-0.34 and 2.54+/-1.25, respectively. All metastatic tumors except one showed intense 11C-methionine uptake in the entire tumor or in the peripheral margin of the tumor. In meningiomas, 11C-methionine uptake showed a variable increase. In conclusion, brain lesions that show hypo- or isometabolism on FDG PET can be detected and differentiated with high sensitivity and good contrast using 11C-methionine PET. 11C-methionine PET can provide additional information when used in combination with FDG PET in the evaluation of these patients. PMID- 11926380 TI - Whole-body biodistribution, radiation absorbed dose, and brain SPET imaging with [123i]5-i-A-85380 in healthy human subjects. AB - The biodistribution of radioactivity after the administration of a new tracer for alpha4beta2 nicotinic acetylcholine receptors (nAChRs), [123I]5-iodo-3-[2(S)-2 azetidinylmethoxy]pyridine (5-I-A-85380), was studied in ten healthy human subjects. Following administration of 98+/-6 MBq [123I]5-I-A-85380, serial whole body images were acquired over 24 h and corrected for attenuation. One to four brain single-photon emission tomography (SPET) images were also acquired between 2.5 and 24 h. Estimates of radiation absorbed dose were calculated using MIRDOSE 3.1 with a dynamic bladder model and a dynamic gastrointestinal tract model. The estimates of the highest absorbed dose (microGy/MBq) were for the urinary bladder wall (71 and 140), lower large intestine wall (70 and 72), and upper large intestine wall (63 and 64), with 2.4-h and 4.8-h urine voiding intervals, respectively. The whole brain activity at the time of the initial whole-body imaging at 14 min was 5.0% of the injected dose. Consistent with the known distribution of alpha4beta2 nAChRs, SPET images showed the highest activity in the thalamus. These results suggest that [123I]5-I-A-85380 is a promising SPET agent to image alpha4beta2 nAChRs in humans, with acceptable dosimetry and high brain uptake. PMID- 11926381 TI - A method for measurement of the uptake patterns of two beta-emitting radionuclides in the same tissue section with a digital silicon detector: application to a study of 89SrCl2 and 153Sm-EDTMP in a dog with spontaneous osteosarcoma. AB - The biological effect of a radiopharmaceutical depends heavily on the heterogeneity of the uptake in the various tissues. A comparative study of two radiopharmaceuticals should therefore include a comparison of the uptake patterns in different tissues. To eliminate the problems caused by variation in kinetics and tumour characteristics between individuals, such a comparison should be based on measured distributions of the radiopharmaceuticals in the same tissue sample. The excellent linearity between activity and counts in images obtained with a digital silicon strip detector allows such distributions to be derived from two autoradiographs acquired at different time points. This method was applied in a comparison of the uptake patterns of 153Sm-EDTMP and 89SrCl2 in sections obtained from a dog with spontaneous osteosarcoma, containing both tumour and normal bone tissues. As the areas of the section were larger than the detector area, the section had to be cut into smaller parts. Images of these were later merged by means of image processing techniques. There were significant differences in the uptake patterns of the two nuclides. In the primary tumour, the uptake of 153Sm was highly heterogeneous, while 89Sr was more uniformly distributed. In trabecular bone, the accumulation of 153Sm was higher than that of 89Sr. In solid cortical bone, 89Sr had the highest uptake. PMID- 11926382 TI - Alterations of the sympathetic nervous system and metabolic performance of the cardiomyopathic heart. AB - Little is known about the effects of altered sympathetic innervation on the efficiency of the cardiomyopathic heart. Thus, it was the aim of this study to determine non-invasively the relationship between the metabolic cost of cardiac work and alterations of the sympathetic nervous system. In ten patients with chronic idiopathic dilated cardiomyopathy, left ventricular function and geometry were assessed by tomographic radionuclide angiography. Myocardial oxidative metabolism was quantified by the clearance constant k(mono) for carbon-11 acetate, determined by positron emission tomography (PET). Cardiac efficiency was then estimated by the work-metabolic index [WMI = stroke volume index x heart rate x systolic pressure/k(mono)]. Additionally, myocardial presynaptic sympathetic innervation was investigated in the same PET session using 11C hydroxyephedrine, and plasma norepinephrine levels were determined by high performance liquid chromatography. The WMI was 3.0+/-1.3 mmHg x ml x 10(6)/m2 and correlated significantly with left ventricular ejection fraction (19%+/-10%; r=0.66, P<0.04) and peripheral vascular resistance (3,638+/-1,085 dynes x s/cm5; r=-0.75, P<0.01). Cardiac hydroxyephedrine retention, reflecting integrity of presynaptic innervation, was abnormally low in 58%+/-38% of the left ventricle. Cardiac hydroxyephedrine retention (r=0.65, P<0.05) and plasma norepinephrine concentration (r=-0.80, P<0.01) both correlated significantly with the WMI. At multivariate stepwise linear regression, only plasma norepinephrine and peripheral vascular resistance were independent determinants of the WMI. In conclusion, metabolic performance of the cardiomyopathic heart decreases with increasing alterations of the sympathetic nervous system. Increasing afterload and effects of systemic sympathetic activation, however, seem to play a dominant role while direct alterations of cardiac presynaptic sympathetic innervation are not independently associated. PMID- 11926383 TI - Quantitative assessment of blood flow reserve using 99mTc-HMPAO in carotid stenosis. AB - Dynamic imaging of the inflow of technetium-99m hexamethylpropylene amine oxime (HMPAO) to the brain has been proved to allow estimation of the hemispherical cerebral blood flow (CBF) using the Patlak plot. In this study, we compared the hemispherical CBF (in ml/min/100 g) of different patient groups. A total of 25 patients (comprising 13 with migraine and 12 scheduled for endarterectomy owing to angiographically confirmed severe stenosis of the internal carotid artery on at least one side) underwent baseline and acetazolamide 99mTc-HMPAO brain perfusion studies. In addition, acetazolamide 99mTc-HMPAO studies were performed in 12 healthy subjects (no baseline study was performed for ethical reasons.) Dynamic studies were acquired by means of a dual-detector gamma camera with a large field of view (HELIX, Elscint). Special difference images were created to make definition of the aortic arch and hemispherical brain regions easier and more reproducible. A semi-automatic method was developed to determine the transit time from the aorta to the brain, making the generation of the Patlak plot even more robust. The baseline CBF values did not significantly depend on the disease (P>0.1), whereas the CBF values obtained after acetazolamide provocation did do so (ANOVA, P<0.001). Patients suffering from migraine showed a significant increase in global CBF values after acetazolamide provocation (paired t test, P<0.05), but we could not find any effect of the provocation in patients awaiting carotid endarterectomy, indicating a lack of cerebrovascular reserve capacity. Comparison of the results of the acetazolamide study in patients and the control group revealed the CBF values to be significantly lower in patients with carotid stenosis (two-sample t-test, P<0.001), but not in those with migraine (P>0.1). In summary, using quantitative analysis of 99mTc-HMPAO brain studies we could objectively compare the CBF of patients suffering from different diseases. Especially the CBF values obtained after acetazolamide provocation permitted effective differentiation of disease states. The quantitative results may be of assistance in therapy planning, e.g. in selection of the correct operative technique. PMID- 11926384 TI - Effective local control of malignant melanoma by intratumoural injection of a beta-emitting radionuclide. AB - Intratumoural injection of an unsealed beta-emitting radionuclide is a new technique for the local control of tumours that has the advantage of delivering a higher radiation dose to tumour while minimising radiation hazard to the surrounding normal tissues. In this study, therapeutic effect, morphological alterations and biological responses to the high-dose continuous irradiation delivered using this new technique were evaluated in an animal model with B16 melanoma. For evaluation of the therapeutic effect, 92 C57BL/6 mice with B16 melanoma were divided into four groups. In each group, intratumoural injections were performed when the tumour measured approximately 1 cm along its long axis. Group 1 (n=25) received 0.3 ml of normal saline, group 2 (n=15) 37 MBq of carrier free holmium-166 in 0.3 ml saline, group 3 (n=27) 185 MBq of 166Ho in 0.3 ml saline and group 4 (n=25) 185 MBq of 166Ho in 0.5 ml saline. In addition, another 30 mice were used for morphological and biological analysis of the radiation effect. These 30 mice were injected with 185 MBq of 166Ho in 0.3 ml saline, and five were sacrificed at each of the following six time points: before injection and 1, 2, 3, 6 and 14 days post injection. Haematoxylin-eosin (H&E) staining, immunohistochemical analysis for p53, p21, PCNA and cyclin D1, terminal deoxynucleotidyl transferase-mediated deoxyuridine triphosphate nick end labelling (TUNEL) staining, reverse transcription-polymerase chain reaction (RT PCR) and flow cytometry were performed. For visual side-by-side comparison, melanoma cells were inoculated bilaterally into the back of ten additional mice, and 185 MBq of 166Ho in 0.3 ml of saline or an equal volume of normal saline was injected separately into the bilateral tumours. Nine days after inoculation of melanoma cells, mean tumour volume reached 492.5-631.9 mm3. Tumours of the control group (group 1) showed rapid growth, and the mean tumour volume reached approximately 30 times the original volume. None of the control group lived for more than 16 days following the injection of normal saline. On the other hand, mean tumour volume of the treated groups showed a gradual decrease, and 67%-74% of the treated animals were alive when all the control animals had died. The median survival of the control group was 9 days following injection, whereas it was 29 days in group 2, 33 days in group 3 and 33 days in group 4. The survival rate of group 3 was higher than that of groups 2 and 4, but statistical significance was not observed. H&E stain of the tumours demonstrated central necrosis and peripheral residual cells with progressive cytoplasmic and nuclear swelling without apoptotic features. Expression of proteins and mRNAs of p53 and bax increased until 3 days, as compared with 48 h for p21; thereafter, the expression gradually decreased. TUNEL-positive nuclei could be seen from 2 days until 2 weeks after treatment. Flow cytometry did not demonstrate an increase in apoptotic features as compared with the control animals. In conclusion, intratumoural injection of the unsealed beta-emitting radionuclide 166Ho appears to be a promising alternative radiotherapeutic modality for the local control of malignant melanoma. The main cell death mechanisms with this technique seem to be radiation-induced central necrosis and peripheral growth arrest or secondary necrosis of tumour cells, rather than apoptosis. PMID- 11926385 TI - Monitoring isotretinoin therapy in thyroid cancer using 18F-FDG PET. AB - Treatment with isotretinoin (13-cis-retinoic acid, 13-cis-RA) is a recent additional option in advanced, otherwise intractable differentiated thyroid cancers. The aim of this study was to evaluate fluorine-18 fluorodeoxyglucose positron emission tomography (18F-FDG PET) in the prediction and the monitoring of response to 13-cis-RA therapy. Twenty-one patients with advanced differentiated thyroid cancers were investigated using 18F-FDG PET and iodine-131 whole-body scans before and 3, 6 and 9 months after initiation of 13-cis-RA therapy. After 9 months, 13-cis-RA treatment was discontinued and imaging procedures repeated 3 months later. Average 18F-FDG uptake (SUV) decreased significantly during 13-cis-RA therapy but subsequently increased in five of eight patients after withdrawal of 13-cis-RA. 18F-FDG uptake (SUV) 3 months after onset of 13-cis-RA therapy was significantly lower in patients who developed increased 131I uptake in their tumour sites than in patients with no subsequent increase in 131I uptake. There was no relationship between serum thyroglobulin level on the one hand and simultaneously measured 131I or 18F-FDG uptake on the other hand. There was a tendency towards lower 18F-FDG uptake in tumour manifestations with a better outcome. Therefore, 18F-FDG PET at 3 months after the start of treatment promises to differentiate between those patients who will eventually benefit from 13-cis-RA and those who will not. In conclusion, these data indicate that 18F-FDG PET is a useful tool for the evaluation and monitoring of adjuvant therapy with 13-cis-RA in thyroid cancer. PMID- 11926386 TI - Non-invasive differentiation of pancreatic lesions: is analysis of FDG kinetics superior to semiquantitative uptake value analysis? AB - The diagnostic utility of fluorine-18 2-deoxy-D-glucose positron emission tomography (FDG PET) for the non-invasive differentiation of focal pancreatic lesions originating from cancer or chronic pancreatitis by combined visual image interpretation and semiquantitative uptake value analysis has been documented. However, in clinical routine some misdiagnosis is still observed. This is because there is potential overlap between the semiquantitative uptake values obtained for active inflammatory lesions and cancer. Therefore, this prospective study was undertaken to test the hypothesis that analysis of dynamic kinetics of focal pancreatic lesions based on FDG PET may more accurately determine the benign or malignant nature of such lesions. Thirty patients (56+/-17 years) were studied dynamically with FDG PET for a period of 60-90 min. Patients were assigned to one of four groups: control, acute pancreatitis, chronic pancreatitis or pancreatic cancer. Two observers, blinded to the clinical data, analysed the time-activity curves of FDG kinetics based on region of interest analysis. The diagnosis predicted by FDG PET was compared with the result of histological examination of the surgical specimen. Analysis of FDG kinetics revealed significant differences in the shape of the time-activity curve for controls, pancreatic cancer and inflammatory disease. Surprisingly, there was no significant difference in the time-activity curve shape for chronic pancreatitis and acute pancreatitis; this is, however, not a clinical issue. Furthermore, acquisition time (60 min vs 90 min) did not affect interpretation of the time-activity curve, so that scanning time may be regularly shortened to 60 min. Interobserver agreement was 1. Based on these findings, non-invasive differentiation between pancreatic cancer and chronic pancreatitis was correctly predicted in all cases, as confirmed by histology. In addition, the specificity was increased compared with that obtained from standardised uptake value analysis. Non-invasive differentiation between pancreatic cancer and chronic pancreatitis may best be achieved based on a dynamic FDG PET study including kinetic analysis. This approach yields results superior to those obtained from a semiquantitative analysis of pancreatic lesions. PMID- 11926388 TI - Effect of postprandial hyperglycaemia in non-invasive measurement of cerebral metabolic rate of glucose in non-diabetic subjects. AB - The aim of this study was to determine the effect of postprandial hyperglycaemia (HG) on the non-invasive measurement of cerebral metabolic rate of glucose (CMRGlc). Five patients who had a meal within an hour before a fluorine-18 fluorodeoxyglucose positron emission tomography (FDG-PET) examination were recruited in this study. They underwent intermittent arterial blood sampling (measured input function), and, based on this sampling, CMRGlc was calculated using an autoradiographic method (CMRGlc(real)). Simulated input functions were generated based on standardised input function, body surface area and net injected dose of FDG, and simulated CMRGlc (CMRGlc(sim)) was also calculated. Percent error of the area under the curve (AUC) between measured (AUC(real)) and simulated input function (AUC(IFsim)) and percent error between CMRGlc(real) and CMRGlc(sim) were calculated. These values were compared with those obtained from a previous study conducted under fasting conditions (F). The serum glucose level in the HG group was significantly higher than that in the F group (165+/-69 vs 100+/-9 mg/dl, P=0.0007). Percent errors of AUC and CMRGlc in grey matter and white matter in HG were significantly higher than those in F (12.9%+/-1.3% vs 3.5%+/-2.2% in AUC, P=0.0015; 18.2%+/-2.2% vs 2.9%+/-1.9% in CMRGlc in grey matter, P=0.0028; 24.0%+/-4.6% vs 3.4%+/-2.2% in CMRGlc in white matter, P=0.0028). It is concluded that a non-invasive method of measuring CMRGlc should be applied only in non-diabetic subjects under fasting conditions. PMID- 11926387 TI - Visualisation and assessment of the protein synthesis rate of lung cancer using carbon-11 tyrosine and positron emission tomography. AB - This study evaluated the potential role of L-(1-(11)C)-tyrosine positron emission tomography (TYR PET) for visualisation and quantification of protein metabolism in lung cancer. Dynamic TYR PET scans of the thorax were performed in 17 patients with lung cancer. Protein synthesis rate (PSR in micromol/min x l) and standardised uptake value (SUV, corrected for body measurements) of tumour tissue and contralateral normal tissue were calculated before and after chemotherapy or radiotherapy. All tumours [11 non-small cell lung carcinomas (NSCLCs), five small cell lung carcinomas (SCLCs), and one pleural mesothelioma] were visualised as a hot spot. The median PSR in tumour tissue was higher than that in corresponding contralateral normal lung tissue before [1.88 micromol/min x l (range 1.10-3.42) vs 0.40 micromol/min x l (range 0.12-0.86); P=0.003] and after treatment [1.33 micromol/min x l (range 0.45-2.21) vs 0.28 micromol/min x l (range 0.18-0.51); P<0.02]. In contrast to PSR of normal lung tissue, PSR of tumour tissue decreased significantly after therapy (P=0.03). Before therapy, no significant difference in PSR between NSCLCs and SCLCs was observed, but after therapy the PSR differed significantly between the subgroups [1.69 micromol/min x l (range 0.63-2.78) for NSCLC vs 0.67 micromol/min x l (range 0.45-0.92) for SCLC; P=0.03], irrespective of the treatment modality. The median SUV of tumour tissue was higher than that in corresponding contralateral normal lung both before and after therapy. Only a weak correlation between PSR and SUV was found when the latter was corrected for body surface area or lean body mass. Carbon-11 labelled tyrosine appears to be a good tracer for visualising lung cancer. PSR of tumour tissue can be used to quantify reduction in the metabolic rate of the tumour. Future studies need to be performed to determine whether TYR PET will supply additional clinical information with treatment implications in patients with lung cancer. PMID- 11926389 TI - Development of technetium-99m-based CNS receptor ligands: have there been any advances? AB - By virtue of its ideal nuclear physical characteristics for routine nuclear medicine diagnostics and its ready availability, technetium-99m is of outstanding interest in the development of novel radiopharmaceuticals. The potential for the development of 99mTc-based radioligands for the study the receptor function in the central nervous system (CNS) is also well recognised despite the difficulties to be overcome. A fundamental challenge is the pharmacologically acceptable integration of the transition metal technetium, with its specific coordination chemistry, into the molecular entity of CNS receptor ligands. Conceptually, the ligand molecule can be assembled by three building blocks: a small neutral chelate unit, an organic linker that may also serve as a pharmacological modifier and a receptor-binding region derived from selective receptor antagonists. The recent introduction of novel technetium chelate units, particularly mixed-ligand complexes and low-valency organometallic compounds of technetium, provides an impetus for the further development of CNS receptor ligands. Moreover, progress in receptor pharmacology and the experience gained with positron emission tomography radiotracers have facilitated the design of numerous 99mTc-based CNS receptor ligands. The formidable challenge of developing 99mTc probes as single photon emission tomography imaging agents targeting CNS receptors can be viewed with optimism given the successful development of [99mTc]TRODAT-1 as a 99mTc complex for imaging dopamine transporters in the brain, although there are a number of receptor-specific imaging agents that have so far resisted all efforts to develop them. This review presents recent advances and discusses the remaining hurdles in the design of 99mTc-based CNS receptor imaging agents. PMID- 11926390 TI - Routine ventilation scans in children with cystic fibrosis. PMID- 11926391 TI - Nephrotoxicity versus anti-tumour efficacy in radiopeptide therapy: facts and myths about the Scylla and Charybdis. PMID- 11926392 TI - Glucose utilisation and cell proliferation in colorectal cancer. PMID- 11926393 TI - A world view at home: the need for bilingualism in the United States. PMID- 11926394 TI - Breastfeeding support. PMID- 11926395 TI - Drug screening and criminal prosecution of pregnant women. AB - According to the U.S. Supreme Court, the Fourth Amendment rights of 10 women were violated by a hospital that provided them prenatal care. The incidence of prenatal drug testing for criminal prosecution with or without a woman's knowledge is increasing. Concurrently, funding and availability of drug treatment programs for pregnant women are declining. Nurses and physicians who act as advocates for the state rather than the patient damage the patient-provider relationship and breach their ethical responsibility to the patient. PMID- 11926396 TI - Neuromotor development and the physiologic effects of positioning in very low birth weight infants. AB - OBJECTIVE: To provide a comprehensive literature review of neuromotor development and related physiologic effects of positioning in very low birth weight infants. DATA SOURCES: MEDLINE, CINHAL, Health Star, Current Contents, and the Australian Medical Index (1966-2000) databases were searched. Unpublished studies (e.g., dissertations, conference proceedings) and all relevant references listed in articles also were examined. STUDY SELECTION: One hundred and eighty theoretical writings, research studies, and clinical papers related to neuromotor development, the physiologic effects of positioning, and interventions to minimize or prevent short- and long-term effects of positioning in very low birth weight infants were reviewed. DATA EXTRACTION: Studies were assessed for scientific rigor, evidence of theoretical foundation, and clinical relevance. Comparisons were made across data sources to determine the most reliable, valid, and consistent findings. DATA SYNTHESIS: Three compelling results emerged: (a) The development of posture and mobility in newborn infants requires an optimal balance between active and passive muscle tone, (b) the prone position is physiologically more beneficial for the preterm infant than supine and lateral positions, and (c) the prone position can lead to short- and long-term postural and associated developmental problems. CONCLUSION: Use of empirically tested postural interventions appropriate for an infant's gestational age, health status, and overall organizational capacity is recommended. PMID- 11926397 TI - A comparison of standardized measures of psychosocial variables with single-item screening measures used in an urban obstetric clinic. AB - OBJECTIVE: To compare standardized, psychometrically sound measures of psychosocial variables and physical violence with single-item measures currently used for screening purposes in an obstetric clinic. STUDY DESIGN: Two hundred multiethnic women were interviewed during scheduled prenatal visits at an urban perinatal center. Participants previously completed a standard perinatal self administered inventory, which included several single-item psychosocial questions. Medical and obstetric history, current risk factors, and intrapartum complications were collected from medical records. RESULTS: Correlational analyses indicated that single-item self-report measures were more appropriate for certain types of variables than for others. Specifically, for anxiety, depression, and social support, single-item measures were significantly correlated with standardized, psychometrically sound measures, whereas this was not true for stress and domestic violence. CONCLUSION: It may be appropriate to rely on self-reported single-item measures to detect some psychosocial risk factors in clinical settings for the purposes of referral. When assessing for domestic violence, single-item measures may not be adequate, as personal interviews using a standardized, psychometrically sound measure resulted in higher rates of reporting. PMID- 11926398 TI - Maternal-fetal attachment in pregnant women who use illicit drugs. AB - OBJECTIVE: To explore cognitive, affective, and altruistic dimensions of maternal fetal attachment in pregnant women who use illicit drugs. DESIGN: Content analysis with deductive and inductive coding methods was used to uncover common themes in each dimension of attachment. SETTING: A prenatal clinic of a tertiary medical center in the northeastern United States. PARTICIPANTS: Forty pregnant women. Inclusion criteria were the following: used illicit drugs after the last menstrual period, had felt fetal movement, and could read and speak English. Women in methadone treatment programs were excluded. RESULTS: Knowing the baby's characteristics and health through fetal movement, acknowledging the fetus as an individual with physical and emotional functions, and knowing the baby by relating the fetus to self and family members are the three major themes in cognitive attachment. Themes in affective attachment include mixing strong affection with guilt and viewing fetal movement as visual and tactile enjoyment but also discomfort. Common themes with conflicting feelings were salient in altruistic attachment, including feeling uncomfortable versus feeling worthwhile, viewing being pregnant as an incentive for lifestyle changes, battling with substance use and concern for fetal health, and alternating between uncertainty and hope in preparing for the baby's arrival. CONCLUSION: Maternal-fetal attachment is not a phenomenon that is present or absent, but a struggle manifested by guilt, concern, and uncertainty. Without proper treatment and counseling, many women struggle between illicit drug use and development of maternal-fetal attachment. PMID- 11926399 TI - Functional status after fatherhood: an Australian study. AB - OBJECTIVE: To investigate the functional status of new fathers. DESIGN: A descriptive, correlational study. SETTING: A variety of postnatal services within one regional center in New South Wales, Australia. PARTICIPANTS: One hundred twenty-eight men who had fathered a healthy infant born at or near term, both first-time fathers and men adding to their existing families, who could read and write English. MAIN OUTCOME MEASURE: Fathers were surveyed at 6 weeks postpartum using the Inventory of Functional Status-Fathers. RESULTS: Paternal age, number of children, and satisfaction with fatherhood were identified as correlates of functional status. An inverse relationship was noted between the number of children and satisfaction with fatherhood. CONCLUSION: Although most fathers maintained their level of participation in household and family activities postpartum, few increased their involvement in response to the birth of their child. Practitioners should encourage expectant parents to actively negotiate the division of household labor and the sharing of infant and child care responsibilities before the birth of their newborn. PMID- 11926401 TI - Human genomics: challenges and opportunities. AB - The pace of knowledge development related to genetics continues to progress exponentially. Knowledge gained through the Human Genome Project will have a profound impact on the practice of nursing, and the widespread integration of genetics into nursing education, clinical practice, and research will be essential. Many nurses continue to view genetic services as relegated to a specialist in a tertiary center and not relevant to their practice. This is certainly about to change. This article reviews the major implications of Human Genome Project for changing the landscape of health care and nursing, discusses several specific examples of genetic testing, and describes strategies under way to help prepare nurses to meet these new demands. Understanding new DNA-based genetic-testing technologies will be important for nurses as these tests become more available and nurses assist clients in the management of their genetic health. Nurses must become active participants in embracing the challenges and opportunities related to this genetic revolution, which over the next 5 to 10 years will reach nurses in every specialty and in every practice setting. PMID- 11926400 TI - Folic acid intake of young women. AB - OBJECTIVE: To investigate the folic acid intake of young women ages 18-24. DESIGN: Nonexperimental, descriptive survey. SETTING: Small private college in western North Carolina. PARTICIPANTS: A convenience sample of 42 female college students ages 18-24. MAIN OUTCOME MEASURES: Survey of demographics, folic acid intake, knowledge of folic acid, and pregnancy contemplation. RESULTS: All participants were found to have an inadequate diet and an inadequate intake of folic acid. Only 33.3% reported taking daily multivitamins. These findings may be attributed to the participants' lack of knowledge about folic acid and the birth defects it helps to prevent. CONCLUSION: Inadequate folic acid intake is common in young women ages 18-24. These young women frequently have poor nutritional habits and are usually noncontemplators of pregnancy. PMID- 11926402 TI - Genetics in perinatal nursing: clinical applications and policy considerations. AB - Genetics is becoming an important part of perinatal nursing care. The need for genetic counseling and referral may be identified before conception, early in pregnancy, upon the demise of a fetus with an abnormality, or after the birth of an infant with a genetic condition. Perinatal nurses often are present when parents first discover the presence of a genetic condition in their fetus or infant. Women who will be 35 at the time of their infant's birth and women who have family histories of genetic disorders should be offered appropriate screening tests. PMID- 11926403 TI - Reproductive options for individuals at risk for transmission of a genetic disorder. AB - The basis of human growth and development has long been considered to be one of the great mysteries of science and mankind. The portal to understanding this mystery was achieved by the Human Genome Project and Celera Genomics in 2001, with their joint announcement of the sequencing of 99% of the human genome map. Current reproductive options, however, remain restricted to the prevention of transmitting an at-risk gene or genes, but do not include treatment or cure. It is anticipated that this state of "halfway technology" will continue for years to come. As such, the scientific and ethical issues associated with each of these reproductive options will continue to affect the decision making of at-risk individuals. As the omnipresent health care provider, nurses have a duty to know and disseminate accurate and current information about reproductive options for individuals at risk for transmission of a genetic disorder. Nurses also have a duty to advocate for and ensure the privacy and confidentiality of genetic information. PMID- 11926404 TI - Newborn screening and genetic testing. AB - Mandated newborn screening programs for genetic and other congenital conditions for the some 4 million infants born in the United States each year have seen dramatic changes over the past decade. With the mapping of the human genome and other advances in science and technology, there will be continued challenges to and changes in these programs. Nurses who care for infants and their families should be knowledgeable about those changes to correctly transmit information to families and to participate in determining policy for newborn screening practices. PMID- 11926405 TI - Cancer genetics and women's health. AB - It is now known that all cancer is genetic in origin. Although most cancer occurs by chance, approximately 5% of individuals inherit specific genetic mutations that predispose them to cancer. The genetic characteristics of some cancers are known. Such information can be useful to health care providers in the clinical setting for treatment, early detection, and prevention. This article reviews basic carcinogenesis as well as genetic syndromes that predispose women to breast, ovarian, colorectal, and endometrial cancer. Knowledge of hereditary cancer syndromes and familiarity with them will assist the advanced practice nurse in the management of patients who are at risk. Cancer continues to contribute substantially to the mortality of women in all age groups. Knowledge of these syndromes provides an excellent opportunity to decrease mortality by early detection and prevention. PMID- 11926406 TI - Heritable cardiovascular disease in women. AB - Although coronary heart disease (CHD) is traditionally viewed as a disease predominantly of males, it is the number one cause of death among American women. Half of all CHD deaths occur in women, and death within the first few years of initial myocardial infarction is significantly higher in women. Few women, however, perceive a significant health risk from heart disease. The dramatic decline in CHD mortality from lifestyle modification suggests that environmental factors play a significant role in genetic expression. Family-focused nursing care affects generations of family members and can significantly reduce morbidity and mortality from CHD. PMID- 11926407 TI - Impaired surface antigen presentation in tumors: implications for T cell-based immunotherapy. AB - The identification of tumor-associated antigens has suggested new possibilities for cancer therapy. However, multiple mechanisms may contribute to the ability of tumor to escape antitumor immune responses. Tumor antigen heterogeneity, modulation of HLA expression and immune suppressive mechanisms may occur at any time during tumor cell progression, and can affect the outcome of therapeutic immune intervention. In particular, the appearance of altered HLA class I phenotypes during tumor development may have important biological and medical implications due to the role of these molecules in T and NK cell functions. Exhaustive tumor tissue studies are necessary before deciding whether a particular patient is suitable for inclusion in T cell-based immunotherapy protocols. PMID- 11926409 TI - HLA class I antigen abnormalities and immune escape by malignant cells. PMID- 11926408 TI - Immunodominance and tumor escape. AB - Cancers in mouse and man express multiple tumor-specific as well as tumor associated antigens. Immunodominance in the host response to these antigens can result in successive selection of heritable antigen loss variants. Immunodominance may also prevent the development of responses to new tumor specific antigens that may arise during tumor progression. Some tumor-specific antigens are retained during tumor progression possibly because they are essential for survival of the malignant phenotype. Immunodominance may allow cancer cells to escape even after loss of a single MHC Class I allele because cross-presentation of the retained antigen by this allele that must be expressed on the surrounding antigen presenting cells sustains the immunodominant response. This prevents effective responses to secondary antigens that may remain as potential targets. Immunization with in vitro selected cancer cell variants that lack the immunodominant antigen can break the immunodominance and prevent escape of cancers from host immunity. PMID- 11926410 TI - Tumour escape from immune surveillance through dendritic cell inactivation. AB - Dendritic cells (DC) are central to the initiation of immunity. To induce immune reactivity, DC are recruited at the site of antigen expression, uptake antigens and migrate to secondary lymphoid organs while receiving activation signals delivered by pathogens, dying cells, and/or T cells. Tumours can escape the immune system by interfering with the migration of DC or by not providing the necessary activation signals. Moreover, tumours promote the secretion of factors that inhibit DC differentiation and functions. We will review the current knowledge of the physiopathology of DC in cancer, which paves the way for novel strategies of therapeutic intervention. PMID- 11926411 TI - Tumor-induced death of immune cells: its mechanisms and consequences. AB - FasL is a death-inducing as well as pro-inflammatory molecule. The seemingly opposing consequences of Fas/FasL interactions appear to be dependent on the context of the tumor or tissue microenvironment. The term 'contextual' has been coined to describe cellular and molecular events that may be opposing, yet are mediated by the engagement of the same receptor-ligand pair. Because Fas is one of the death-inducing receptors, its ligand is dangerous. Hence, the availability of FasL for cross-linking of the receptor has to be extremely carefully regulated. Different levels of control appear to exist, some involving cell membrane-associated MMPs and others depending on cytokines released from neighboring cells. In tumors, which shape their microenvironment to make it unwelcome to immune effector cells, FasL is used to kill infiltrating cells without damage to the tumor, which develops multiple means of hiding/inhibiting the death pathway. On the other hand, in situations provoking inflammatory responses such as allograft rejection, infection, and tissue necrosis, FasL assumes the role of attractant for inflammatory cells. In these situations, FasL acts in conjunction with other cytokines, and its effects might be modified in the context of unfolding events. When put in the context of biologic interactions that take place at the tumor site, the Fas/FasL pathway emerges as a molecular mechanism, which contributes to immune privilege and tumor escape, but which can also enhance anti-tumor inflammatory responses. FasL, like many cytokines, is pleiotropic. It is also worth bearing in mind that other molecular players, including the other known members of the TNF family, almost certainly participate in establishing immune privilege status of tumors. While a broad body of new information has contributed to clarifying the dual role of FasL in the tumor biology, much remains to be learned about molecular mechanisms that control its effects in the tumor and in normal tissues. PMID- 11926412 TI - Defective death receptor signaling as a cause of tumor immune escape. AB - Death receptors are a subgroup of TNF-receptor family members that can trigger caspase-8 activation and apoptosis upon interaction with their selective ligands. One of the death receptors, Fas (CD95) and its ligand is critically involved in the regulation of immune homeostasis and effectorfunction. Fas-mediated cell death is a major pathway of cytolytic T-cell-mediated death that is involved in specific killing of tumor cells. Recent investigations summarized herein have shown that defective Fas-signaling due to receptor downregulation or dysfunction, or intracellular inhibition by FLIP (FLICE inhibitory protein) can interfere with Fas-mediated tumor cell death, and thereby favor tumor immune escape. PMID- 11926413 TI - Tumor cell recognition by natural killer cells. AB - Natural killer (NK) cells contribute to the immune defense against cancer and viruses. Tumor cells and infected cells that downregulate the HLA class I antigen expression are targets for NK cell responses because NK cell activation is controlled by a repertoire of inhibitory receptors with different HLA class I specificities. The clonal distribution of these inhibitory receptors permits NK cell recognition of target cells that have lost expression of a single HLA-B or HLA-C allotype. Several activation receptors on NK cells have been identified that contribute to tumor cell recognition. One such receptor, NKG2D, is expressed by all NK cells and binds to inducible ligands on tumor cells. PMID- 11926414 TI - Strategies to overcome immune ignorance and tolerance. AB - Cancer poses a difficult problem for immunotherapy because it arises from the host's own tissues. Many of the target antigens are tissue-specific molecules shared by cancer cells and normal cells. Thus, these are weak antigens that do not typically elicit immunity. In addition, tumors have several features that make their recognition and destruction by the immune system difficult. Despite these obstacles, several strategies for developing effective tumor immunity have been developed. Crucial to these approaches is the discovery and understanding of the molecular identity of antigens and the mechanisms involved in tumor immunity. In this review, strategies to overcome immune ignorance and tolerance are discussed. PMID- 11926415 TI - Tumor immunology--towards a paradigm of reciprocal research. AB - Recent advances in the field of tumor immunology highlight the difficulties involved in generating and maintaining a tumor-specific immune response. The tendency of T cells to be tolerized in vivo, and the tendency of tumors to escape immune recognition represent significant barriers to successful immunotherapy. The results of early clinical trials illustrate these points and underscore the critical importance of an interactive dialog between laboratory and clinical research efforts. PMID- 11926417 TI - Spinal cord stimulation facilitates functional walking in a chronic, incomplete spinal cord injured. AB - DESIGN: This paper describes a treatment paradigm to facilitate functional gait in a quadriplegic, ASIA C spinal cord injured (SCI), wheelchair-dependent subject who presented with some large fiber sensation, sub-functional motor strength in all lower limb muscles, and moderate spasticity. The study utilizes partial weight bearing therapy (PWBT) followed by epidural spinal cord stimulation (ESCS) with the assumption that both treatments would be necessary to elicit a well organized, near effortless functional gait with a walker. Function is defined in terms of accomplishing task-specific activities in the home and community. OBJECTIVES: To demonstrate the feasibility and benefits of combined PWBT and ESCS therapies aimed at promoting functional gait in a wheelchair-dependent ASIA C SCI subject. SETTING: The Clinical Neurobiology and Bioengineering Research Laboratories at Good Samaritan Regional Medical Center, Phoenix, Arizona, USA, and the Department of Bioengineering, Arizona State University, Tempe, Arizona, USA. METHODS: The study began with the application of PWBT. The subject walked on the treadmill until a plateau in gait rhythm generation was reached. Subsequently, ESCS, applied to the lumbar enlargement, was utilized to facilitate PWBT and, later, over-ground walking for a standard distance of 15 m. Gait performance was analyzed by measuring average speed, stepping symmetry, sense of effort, physical work capacity, and whole body metabolic activity. RESULTS: PWBT led to improved stereotypic stepping patterns associated with markedly reduced spasticity, but was insufficient for over-ground walking in terms of safety, energy cost, and fatigue. ESCS with PWBT generated immediate improvement in the subject's gait rhythm when appropriate stimulation parameters were used. When compared to the non-stimulated condition, over-ground walking with ESCS across a 15 m distance was featured by a reduction in time and energy cost of walking, sense of effort, and a feeling of 'lightness' in the legs. After a few months of training, performance in speed, endurance, and metabolic responses gradually converged with/without ESCS at this short distance, suggesting a learned response to these conditions. However, at longer distances (eg, 50-250 m), performance with ESCS was considerably superior. The subject was able to perform multiple functional tasks within the home and community with ESCS. CONCLUSION: We propose that ESCS augments the use-dependent plasticity created by PWBT and may be a valuable adjunct to post-SCI treadmill training in ASIA C subjects. We also conclude that ESCS elicits greater activation of an oxidative motor unit pool, thereby reducing the subject's sense of effort and energetic cost of walking. PMID- 11926416 TI - Assumptions of the tumor 'escape' hypothesis. AB - The reasons why cancer cells are not destroyed by the immune system are likely to be similar, in most cases, to the reasons why normal cells are not destroyed by the immune system. Unfortunately for tumor immunologists, these reasons have not yet been fully elucidated. What is known, however, is that the lack of autoimmune destruction of normal tissue after immune activation is a finely regulated, highly orchestrated sequence of events. Viewed in this light, it is interesting to conceptualize the derangement of the tumor genome not merely as an engine that enables cancer cells to dodge immune recognition. The dysregulation characteristic of the transformed genome is also what makes tumor immunity, a specialized form of autoimmunity, possible. PMID- 11926418 TI - Functional ability, perceived exertion and employment of the individuals with spinal cord lesion in Taiwan. AB - STUDY DESIGN: A survey on functional ability, perceived exertion, and employment was mailed to 190 individuals with spinal cord lesion. Analysis is based on the 91 completed replies. OBJECTIVES: To establish a database of individuals with spinal cord lesions and to assess the effectiveness of training programs designed to improve life quality and employment chances among the individuals with spinal cord lesion. SETTINGS: Subjects selected from the Spinal Cord Injury Association of the Republic of China (SCIAROC) and from the Asylum Center Spinal Cord Injury (ASCCI) in Taiwan. METHODS: Information was obtained from a survey sent to the subjects who are chosen randomly from the SCIAROC and from all the subjects through ACSCI training program. RESULTS: Respondents in ACSCI had a higher functional activity level and lower perceived exertion during activities than those in SCIAROC. The employment status is related to the functional independence and level of injury. CONCLUSION: The database provides information on functional and employment status of individuals with spinal cord lesion. The present study also demonstrates the efficacy of a vocational training program for individuals with spinal cord lesion in Taiwan. PMID- 11926419 TI - The prevalence and association of neck (coat-hanger) pain and orthostatic (postural) hypotension in human spinal cord injury. AB - OBJECTIVE: To investigate the prevalence of orthostatic (postural) hypotension (OH) and neck pain in a 'coat-hanger' occipito-cervical distribution in subjects with spinal cord injury (SCI), and their association. METHOD: Blood pressure was measured during head-up tilt to 60 degrees (to determine OH) and neck pain was assessed in 28 subjects with SCI (cervical, thoracic and lumbar level) with McGill Pain Questionnaire, visual analogue scale for pain intensity and Orthostatic Intolerance Symptoms Questionnaire for pain frequency. RESULTS: Neck pain was reported by 53.6% of subjects. Orthostatic hypotension was present in 57.1% of subjects. Neck pain was reported by 75% of subjects with OH and 25% of subjects without OH (P<0.03, Chi-square). Features of such pain included positive correlation to upright posture and exercise, and relief when lying flat. CONCLUSION: There is a high prevalence of neck pain and OH in SCI, with a positive association similar to that reported in primary autonomic failure with OH. PMID- 11926421 TI - Late superior mesenteric artery syndrome in paraplegia: case report and review. AB - STUDY DESIGN: A case report of superior mesenteric artery syndrome (SMA syndrome) occurring in a paraplegic patient 3 months after injury. OBJECTIVES: To report an unusual case and review the literature of SMA syndrome in spinal cord injured patients, focusing on paraplegic subjects and on tardive presentations. SETTING: A Physical Medicine and Rehabilitation Center in Garches (France). METHOD: Current medical literature includes reports of only 14 spinal cord injured patients with SMA syndrome. This syndrome has been often described in anorexia nervosa, burns or other causes of cachexia, following correction of spinal deformities or after application of body casts. RESULTS: In spinal cord injured patients SMA usually occurs in tetraplegic patients during the first weeks after injury. Only four cases of SMA syndrome in paraplegic patients have been described. Late forms are less common than acute ones: only three cases among 14. SMA syndrome consists of a vascular compression of the third part of the duodenum between the ventrally oriented SMA and the aorta. The normal aorto-mesenteric angle ranges between 38 degrees and 65 degrees and can be as low as 6 degrees in patients with SMA syndrome. The diagnosis is usually based on upper gastro intestinal contrast X-ray study, which shows abrupt vertical compression of the third part of the duodenum. CT scan with angiography is useful in some difficult cases. Conservative management includes early correction of dehydration and electrolyte imbalance, insertion of a nasojejunal tube beyond the obstruction and renutrition. Duodenojejunostomy may be necessary in case of failure of conservative treatment. CONCLUSION: SMA syndrome is an unusual gastro-intestinal complication that may occur in paraplegic patients, even late after injury. PMID- 11926420 TI - Ultrasound bone mass in paraplegic patients. AB - STUDY DESIGN: A cross-sectional study. OBJECTIVE: To clarify the existing controversy with regard to whether paraplegic patients suffer a loss of bone mass in the upper limbs. SETTING: Madrid, Spain. METHODS: We evaluated bone mass by phalangeal ultrasonography in 35 male patients with paraplegia (mean age 49+/-12 years), and 25(OH)D3 and PTH to exclude the presence of osteomalacia and secondary hyperparathyroidism. Spasticity was evaluated according to the Ashworth scale. Patients were compared with a control group of 35 healthy male subjects (mean age 48+/-13 years). RESULTS: The patients had lower 25(OH)D3 levels and amplitude-dependent speed of sound (Ad-SOS) than controls (both P<0.001), and higher PTH levels (P<0.05). There was a statistically significant negative association between PTH and 25(OH)D3 levels (r=-0.52, P<0.0001, CI -0.73 to 0.22) and between 25(OH)D3 and injury duration (r=0.34, P<0.05, CI -0.60 to 0.01). There was no correlation between Ad-SOS values, levels of PTH or 25(OH)D3, and the injury duration. No significant difference in Ad-SOS values was found in patients grouped according to low-to-normal 25(OH)D3 level or according to normal to-high PTH level. There were no differences in relation to muscle tone. Only alkaline phosphatase and tartrate-resistant acid phosphatase levels were higher in patients than in controls (both P<0.001). CONCLUSION: Paraplegic patients had a loss of phalangeal bone mass that was unrelated to the levels of vitamin D or PTH, or to muscle tone, so it seems to be related to increased bone resorption rather than to deficient bone formation. PMID- 11926422 TI - Mid-line clefts of the atlas: a diagnostic dilemma. AB - Congenital clefts and other developmental anomalies of the atlas vertebra are rarely encountered. They are incidental findings discovered while investigating the cervical spine following trauma. Differentiation of developmental variants of the atlas from the burst fracture of Jefferson is essential to prevent unnecessary medical intervention. PMID- 11926423 TI - Extra corporeal shock wave lithotripsy of calculi located in lower calyx of left kidney in a spinal cord injury patient who has implantation of baclofen pump in the ipsilateral loin. PMID- 11926424 TI - Pain following spinal cord injury. PMID- 11926425 TI - On use and purpose in the sociology of CAM. PMID- 11926426 TI - Nurses' use of professional distancing in the appropriation of CAM: a text analysis. AB - Despite the depth of interest in complementary and alternative medicines (CAM) within the nursing community, the intersection between nurses, nursing and CAM has largely avoided sociological analysis. This paper presents findings from one part of an ongoing exploratory study of how nurses present and interpret the apparent affinity of their profession with CAM. A text analysis of papers published from within the nursing community on CAM was conducted. Within the broad area of professional identity, differentiation and development, two dominant themes emerged from the analysis. Firstly, the use of professional distancing to underpin the legitimacy of the nursing/CAM axis; and second, the potential offered by the relationship for a reconstruction of lines of professional authority. As one dimension of the way in which nursing's intersection with CAM is being constructed and perpetuated, the relationship with medicine appears to be of significance and requires further attention. PMID- 11926427 TI - Test-retest-reliability and validity of the Kinesiology muscle test. AB - OBJECTIVES: To assess the test-retest-reliability and validity of the Health Kinesiology muscle test. PATIENTS: Seven patients with clinically and allergologically confirmed wasp venom allergy. DESIGN: Four Health Kinesiology examiners tested each patient in a random order for 10 verum and 10 placebo bottles. All examiners used the anterior deltoid as indicator muscle. Patients and examiners were completely blinded. OUTCOME MEASURES: Weak muscle holds were rated as 'sensitivity' towards the test substance, stable holding as normal (not sensitive). RESULTS: An overall kappa of 0.03 (95%-CI: -0.02-0.07) indicates the test is not reliable. Individual kappas do not substantially vary from examiner to examiner. Sensitivity and specificity were estimated at 40% and 60%. CONCLUSIONS: The results suggest that the use of Health Kinesiology as a diagnostic tool is not more useful than random guessing. This should at least be true in patients with insect venom allergy that are tested by examiners with average skills. PMID- 11926428 TI - How traditional Chinese medicine acupuncturists would diagnose and treat chronic low back pain: results of a survey of licensed acupuncturists in Washington State. AB - OBJECTIVES: This survey was undertaken to learn how Traditional Chinese Medicine acupuncturists' diagnose and treat patients with chronic low back pain in order to develop a standardized treatment for a clinical trial of that condition. DESIGN: We surveyed a randomly selected group of 56 acupuncturists in Washington State, USA about styles of acupuncture they used for treating chronic low back pain, diagnoses made, and key features of treatment for this condition. RESULTS: While substantial variability existed among practitioners, there was agreement on several broad features of treatment including: the use of local and distal acupuncture points (86% of practitioners), the use of acupuncture points on the meridians traversing the back (especially the UB meridian, 90%) the use of acupoints determined by palpation (82%), the importance of eliciting de qi (60%), and of providing up to eight treatments for achieving therapeutic results (79%). CONCLUSION: The use of practitioner surveys can enhance the systematic development of acupuncture treatment protocols and should be part of this process in future clinical trials of common conditions. PMID- 11926429 TI - Variation in diagnosis and treatment of chronic low back pain by traditional Chinese medicine acupuncturists. AB - OBJECTIVES: To assess interrater reliability of Traditional Chinese Medicine (TCM) diagnosis and treatment of chronic low back pain. DESIGN: Under a Latin square design, six TCM acupuncturists evaluated the same six patients on the same day. SETTING: Northwest Institute of Acupuncture and Oriental Medicine, Seattle, Washington. INTERVENTIONS: Assessment only. OUTCOME MEASURES: TCM diagnosis, acupoint prescriptions, auxiliary treatment recommendations. RESULTS: Twenty diagnoses and 65 acupoints were used at least once. The diagnosis of Qi/Blood Stagnation with Kidney Deficiency and the acupoint UB23 were used for every patient by most acupuncturists. However, consistency across acupuncturists regarding diagnostic details and other acupoints was poor. No diagnoses, and only one acupoint, were used preferentially for a subgroup of patients. Some diagnoses and treatment recommendations were dependent more on the practitioner than on the patient. Fine-grained diagnoses and most acupoints were unrelated to either patient or practitioner. CONCLUSIONS: TCM diagnoses and treatment recommendations for specific patients with chronic low back pain vary widely across practitioners. Acupuncture clinical trials using an individualized treatment arm may be difficult to replicate or evaluate because of low concordance among acupuncturists. Comparison of individualized treatment with a thoughtfully developed standardized approach is warranted to determine which, if either, is superior. PMID- 11926430 TI - Attitudes to and use of complementary medicine among physicians in the United Kingdom. AB - OBJECTIVES: To evaluate the use of attitudes to complementary and alternative medicine (CAM) among UK physicians. DESIGN: Postal questionnaire. SUBJECTS: All Members and Fellows of the Royal College of Physicians. RESULTS: Twelve thousand, one hundred and sixty eight Members and Fellows were surveyed and a response rate of 23% (n = 2,875) was obtained. Responses from the small numbers of general practitioners (n = 127) were not included in the analyses, resulting in a sample size of 2,748: 79% of respondents were in NHS practice, 32% of respondents practised CAM themselves, and 41% referred patients to CAM; of those who referred patients, 78% referred between 0-3 patients per month. CAM is used by physicians more frequently in private as compared to NHS practice. Acupuncture, aromatherapy and manipulative medicine (osteopathy and chiropractic) are the most commonly referred to and the most commonly practised therapies. Eighty seven percent of those using CAM themselves, or as part of their clinical team's commitment, had not had any CAM training. Attitudes to CAM were generally positive, particularly among those in palliative care, rehabilitation, nuclear medicine, and genito urinary medicine. CONCLUSIONS: Our conclusions must be tempered by the limited response rate, but assuming all non-responders were disinterested in CAM, then at least one in ten UK specialist physicians are actively involved in CAM treatments, although only 13% of our sample had received any CAM training. PMID- 11926431 TI - Physician use of complementary and alternative medicine (CAM) literature. AB - OBJECTIVE: To document the interest of complementary and alternative medicine (CAM) literature use among physicians. DESIGN: A retrospective study. SETTING: A medical library service. MAIN OUTCOME MEASURES: Analysis of photocopy requests from published lists of reference in a two-year period by physicians in the province of British Columbia, Canada. RESULTS: The average number of requests per article was 18.7 among CAM literature and 7.1 among non-CAM literature. Ten of the 15 most frequently requested articles were CAM papers. Review articles on herbal medicine demonstrated the most use. CONCLUSIONS: There is a definite interest in CAM literature among a specific group of Canadian physicians. Use of scientifically credible literature in peer-reviewed journals will be helpful for patient care, teaching, and research in this field. PMID- 11926432 TI - Which complementary and alternative therapies benefit which conditions? A survey of the opinions of 223 professional organizations. AB - With the increasing demand and usage of complementary/alternative medicine (CAM) by the general public, it is vital that healthcare professionals can make informed decisions when advising or referring their patients who wish to use CAM. Therefore they might benefit from advice by CAM-providers as to which treatment can be recommended for which condition. AIM: The primary aim of this survey was to determine which complementary therapies are believed by their respective representing professional organizations to be suited for which medical conditions. METHOD: 223 questionnaires were sent out to CAM organizations representing a single CAM therapy. The respondents were asked to list the 15 conditions they felt benefited most from their CAM therapy, the 15 most important contra-indications, the typical costs of initial and any subsequent treatments and the average length of training required to become a fully qualified practitioner. The conditions and contra-indications quoted by responding CAM organizations were recorded and the top five of each were determined. Treatment costs and hours of training were expressed as ranges. RESULTS: Of the 223 questionnaires sent out, 66 were completed and returned. Taking undelivered questionnaires into account, the response rate was 34%. Two or more responses were received from CAM organizations representing twelve therapies: aromatherapy, Bach flower remedies, Bowen technique, chiropractic, homoeopathy, hypnotherapy, magnet therapy, massage, nutrition, reflexology, Reiki and yoga. The top seven common conditions deemed to benefit by all twelve therapies, in order of frequency, were: stress/anxiety, headaches/migraine, back pain, respiratory problems (including asthma), insomnia, cardiovascular problems and musculoskeletal problems. Aromatherapy, Bach flower remedies, hypnotherapy, massage, nutrition, reflexology, Reiki and yoga were all recommended as suitable treatments for stress/anxiety. Aromatherapy, Bowen technique, chiropractic, hypnotherapy, massage, nutrition, reflexology, Reiki and yoga were all recommended for headache/migraine. Bowen technique, chiropractic, magnet therapy, massage, reflexology and yoga were recommended for back pain. None of the therapies cost more than l60 for an initial consultation and treatment. No obvious correlation between length of training and treatment cost was apparent. CONCLUSION: The recommendations by CAM organizations responding to this survey may provide guidance to health care professionals wishing to advise or refer patients interested in using CAM. PMID- 11926433 TI - Research priorities in CAM. PMID- 11926434 TI - CAM evaluation: what are the research questions? AB - Much effort is being directed towards the evaluation of CAM therapies. Yet evaluation is often encouraged before truly considering the research questions which need to be asked. Research questions like 'Does this CAM therapy work?' must be clearly distinguished from 'Does this particular CAM treatment work?' and the latter question needs to be replaced by even more specific ones like 'Does it work when administered by therapist X?' and 'Is therapist X effective?'. Suggestions for the study of the therapeutic process and the evaluation of therapist effectiveness are presented. PMID- 11926435 TI - The term 'complementary medicine' is no more satisfactory as a collective definition of the disciplines that many of us practice than the older adjectives of 'alternative' or 'fringe'. PMID- 11926436 TI - Economic evaluation of linezolid, flucloxacillin and vancomycin in the empirical treatment of cellulitis in UK hospitals: a decision analytical model. AB - Standard antibiotic treatment of infections has become more difficult and costly due to treatment failure associated with the rise in bacterial resistance. New antibiotics that can overcome such resistant pathogens have the potential for great clinical and economic impact. Linezolid is a new antibiotic that is effective in the treatment of both antibiotic-susceptible and antibiotic resistant Gram-positive bacterial infections, including those resistant to other available antibiotics. This breadth of activity is unique in existing antibiotics for Gram-positive bacteria and serves as the rationale for exploring the hypothesis that linezolid is an appropriate choice when considering empirical treatment of cellulitis in complicated or compromised patients in the nosocomial setting. A decision-modelling approach was used to compare the predicted first line treatment efficacy and direct medical costs of linezolid with standard treatment of cellulitis among hospitalized patients. For the purposes of this analysis, standard care is defined along two main pathways: (1) initiating care with intravenous (iv) flucloxacillin, switching to vancomycin if the pathogen is found to be resistant to flucloxacillin, or maintaining flucloxacillin if the pathogen is found susceptible, or when culture and sensitivity analysis is inconclusive; or (2) initiating care with vancomycin, switching to iv flucloxacillin if the pathogen is found susceptible to flucloxacillin, maintaining vancomycin if the infection is found resistant, or when culture and sensitivity are inconclusive. For those patients taking iv flucloxacillin, a switch to oral flucloxacillin was allowed when clinically appropriate. We hypothesized that the cost of care of initiating treatment with linezolid would be less than that for both vancomycin and flucloxacillin in resistance risk ranges typically encountered in UK hospitals. In addition, while the registration trials showed equivalence of linezolid with the comparators in known or suspected methicillin-resistant Staphylococcus aureus (MRSA) and in known or suspected methicillin-susceptible Staphylococcus aureus (MSSA) (vancomycin and oxacillin) respectively, we hypothesized that first-line success rates would be higher in empiric treatment with linezolid. Efficacy data were obtained from recent clinical trials with linezolid and standard treatment, and medical resource utilization was obtained from an expert panel of clinicians who were questioned regarding resistant and susceptible infections separately. UK hospital direct medical costs of treatment were determined using standard costing techniques. Base case analyses assumed a residual 80% unknown pathogen rate after culture and susceptibility based on a physician survey and supported in the literature. The analysis in this model predicts that initiating empirical treatment of cellulitis with linezolid will (1) result in higher overall success rates than flucloxacillin for first-line treatment, regardless of resistance risk and (2) be less costly than initiating treatment with flucloxacillin when the likelihood of a patient being infected by a resistant pathogen is greater than 24.1%. Furthermore, initiating treatment with linezolid is predicted to result in higher overall success rates and be less costly than vancomycin across the entire spectrum of the patients' risk of being infected by a resistant pathogen. PMID- 11926437 TI - Therapeutic options for Gram-positive infections. AB - Gram-positive infections impose a major burden on patients and the healthcare systems globally. The need to treat these infections correctly in an empirical fashion is of paramount importance. Further complicating this changing aetiology is the emergence of resistant strains which are no longer predictably susceptible to standard first-line antimicrobials such as oxacillin or vancomycin. Thus new agents such as linezolid have been developed to assist with initial empirical prescribing in infections where Gram-positive pathogens may be present. The characteristics of linezolid, including spectrum of activity, pharmacodynamic profile, tolerablility and overall efficacy should strengthen confidence when considering initial antimicrobial therapy in patients in risk areas. Future agents also being developed to fight multi-resistant Gram-positive infections include oritavancin, daptomycin and the glycylcyclines; however, these are still in the development phase. PMID- 11926438 TI - Antimicrobial susceptibility of Gram-positive bacteria: what's current, what's anticipated? AB - Changing patterns of pathogens and antibiotic susceptibility present clinicians with difficult choices for antimicrobial prescribing. In particular, multiresistant staphylococci, enterococci and pneumococci present problems in many settings. The number of predictably active antimicrobials is decreasing in many centres, with significant consequences for both patients and society as a whole. New antimicrobial options have been few in recent years and several promising quinolones have been compromised by formulation and/or toxicity issues. Nevertheless, the recent introduction of linezolid and quinupristin/dalfopristin provides clinicians with valuable new options against Gram-positive cocci. These options should further increase with the likely introduction of daptomycin, oritavancin and tigilcycline. A range of surveillance programmes helps monitor the ever-changing patterns of resistance and thus guides clinicians in their empirical prescribing. Empirical use of powerful newer agents may be justifiable in seriously ill patients in those settings, units and countries where there is a substantial background rate of resistance. PMID- 11926439 TI - Economic impact and formulary positioning of linezolid: a new anti-Gram-positive antimicrobial. AB - Gram-positive bacteria have emerged as major causes of colonization and serious infection within the nosocomial and increasingly also within the community setting. These infections have significantly contributed to patient morbidity and mortality as well as prolongation of hospital stay, a key determinant of the cost of an episode of infection in hospital. In many countries globally, infections due to methicillin-resistant Staphylococcus aureus (MRSA) are providing the greatest burden of clinical infection, often occurring in vulnerable patients or "high risk" therapeutic settings. Combined with this scenario is the increasing requirement for health care organizations to provide cost-effective health care as well as care that is delivered on evidence-based practice delivered through formularies or guidelines. This article aims to: 1) summarize the key economic considerations pertinent to these multiresistant infections but with an emphasis on MRSA, 2) discuss the current therapeutic options of managing MRSA infections, and 3) discuss the formulary positioning of linezolid by means of outlining its core strengths, weaknesses and the opportunity it provides to hospital infection management. PMID- 11926440 TI - Qualitative findings on the experience of end-of-life care for hematological malignancies. AB - There is a loud silence in the literature on the issues related to palliative care for hematological malignancies. This article presents information that begins to address that silence. The limited research that is available indicates that, to date, patients with a hematological malignancy and their families received scant attention with regards to the provision of palliative care. The findings presented in this article demonstrates that families are enduring considerable hardship and are left with much emotional pain and many unresolved issues when such care is not offered. It is hoped that the findings presented in this article will be used for developing effective strategies to ensure that patients with hematological malignancies and their families, no less than any others, can be afforded the dignity and respect that the appropriate provision of palliative services ensures. PMID- 11926442 TI - Redefining hope for the terminally ill. AB - This paper discusses how hospice social workers assist patients and families in finding new hope and meaning in their lives as care goals turn from cure to comfort. Assessment factors important to the redefinition process and intervention strategies are explored. Hope is defined as the positive expectation for meaning attached to life events with the emphasis on meaning instead of life events. The author seeks to demonstrate the importance of meaning to the feeling of hope and the possibility of socially constructing meaning to alleviate the traditional medical perspective that hope revolves around the outcome of disease. PMID- 11926441 TI - Is it appropriate to screen palliative care patients for depression? AB - It is estimated that approximately 25 percent of palliative care patients have symptoms of depression, but much of this depression is not identified and therefore not treated. Reasons for nonidentification include the difficulties of distinguishing between what can be called "appropriate sadness" and depression at the end of life and also the nondisclosure by patients of their own mood. In an effort to improve the early detection of depression, patients of all age groups referred to a clinical nurse specialist team within a six-month period were invited to complete the Edinburgh Postnatal Depression Scale (EPDS); the scale was found in an earlier study to have a sensitivity and specificity of above 80 percent at a cutoff threshold of 13 or above. The present study found that 34 percent of patients scored at or above the previously validated threshold of 13, and that younger patients (under age 50) were twice as likely to score above the threshold than were older patients (over age 70). The scale was easy to complete by patients, and staff found it useful as part of their initial assessment of patients. It is suggested that such a tool may aid the early detection and treatment of depression in palliative care patients. PMID- 11926443 TI - Perspectives of relatives and health care workers on care of terminally ill patients in the UK. AB - Two parallel questionnaires were used to explore perceptions of healthcare workers (HCWs) and the relatives of terminally ill patients on the quality of care received by the patients. There was general agreement between the two groups of respondents in most of the assessed areas. The relatives, however were more satisfied with the control of pain and psychological symptoms than the HCWs, and the HCWs perceived the control of diarrhea, skin problems, and swallowing problems more positively than did the relatives. There also was some variation between the two groups in their perception of the care received by the relatives during visits at the hospital, with the relatives' group being more positive. Although HCWs reported that time to deal with patients and their families was inadequate, the relatives' group was generally satisfied with the standards in place. PMID- 11926444 TI - Creating a hospice pharmacy and therapeutics committee. AB - Implementing a Pharmacy and Therapeutics Committee (P&T) as a management strategy for Hospice of the Bluegrass in Lexington, Kentucky, has proven to be effective in reducing costs and improving patient outcomes. Early efforts of the committee yielded the establishment of protocols and guidelines, educational programs, pharmacy newsletters for nurses, and patient education material. In the spring of 2000, Hospice of the Bluegrass developed a preferred drug list (PDL) consisting of the medications it considered essential for effective pain and symptom control. The addition of a clinical pharmacist and a P&T committee has resulted in significant cost savings and improved pharmacotherapeutic care for patients of Hospice of the Bluegrass. This model is an option for any hospice looking to achieve the same outcomes. PMID- 11926445 TI - Improving efficacy, efficiency and economics of hospice individualized drug therapy. PMID- 11926446 TI - Babes of innocence. PMID- 11926447 TI - Dying from hematological cancers. PMID- 11926448 TI - Palliative sedation vs. terminal sedation: what's in a name? PMID- 11926449 TI - Post-thoracotomy pain syndrome: an opportunity for palliative care. PMID- 11926450 TI - The business of palliative medicine--Part 2: The economics of acute inpatient palliative medicine. AB - Economic feasibility is a major factor in the viability of established acute inpatient palliative medicine. Several clinical, administrative, and financial parameters determine the financial health of inpatient care. Financial management metrics include case mix index (CMI) (as determined by the Federal Register as an assigned relative weight to the diagnosis-related group (DRG) reflecting resource consumption), direct costs, indirect costs, contribution margin, and in the future of all patient revised-DRG (APR-DRG). Both census and length of stay will have a major impact on these financial metrics. The type of patient referral and clinical decisions will influence direct costs and revenues. In the future, an international CMI or APR-DRG will allow palliative inpatient units to compare disease severity, resource utilization, and outcome measures. PMID- 11926451 TI - Ergonomic and demographic issues reported by palliative care workers in southern Taiwan. AB - AIM: To investigate ergonomic and demographic issues reported by palliative care workers in southern Taiwan. METHOD: A structured questionnaire was completed by staff members from 11 nursing homes within southern Taiwan. Questions focused on age, sex, height, weight, shift-work details, duration of current employment, nature of current employment, the presence of injury and pain during the past 12 months, the phase lag before the onset of injury, and sick leave details. RESULTS: A total of 125 health-care workers were recruited. Most were female (89.3 percent) and employed full-time (64.8 percent) as patient care assistants (55.2 percent). Most workers were regularly involved in patient-handling tasks (61.6 percent). Almost half (36.8 percent) had suffered a musculoskeletal disorder within the previous year. Lower back injury was reported by 12.0 percent of all employees. Changing patients'clothes and changing their bed linen were associated statistically with musculoskeletal injury (odds ratio (OR) 2.9, 95 percent confidence interval (CI) 1.4-6.3 and OR 2.8, 95 percent CI 1.4-6.1), respectively. Moving the patients was also related to injury during our research (OR 2.5, 95 percent CI 1.2-5.4). CONCLUSION: This study has revealed various ergonomic and demographic issues reported by palliative care workers in southern Taiwan for the first time. The prevalence of certain injuries and symptoms are different from other reports. PMID- 11926452 TI - Galton's legacy to research on intelligence. AB - In the 1999 Galton Lecture for the annual conference of The Galton Institute, the author summarizes the main elements of Galton's ideas about human mental ability and the research paradigm they generated, including the concept of 'general' mental ability, its hereditary component, its physical basis, racial differences, and methods for measuring individual differences in general ability. Although the conclusions Galton drew from his empirical studies were seldom compelling for lack of the needed technology and methods of statistical inference in his day, contemporary research has generally borne out most of Galton's original and largely intuitive ideas, which still inspire mainstream scientific research on intelligence. PMID- 11926453 TI - The effect of the health care supply environment on children's nutritional status in rural Nepal. AB - This article examines the effect of access to health infrastructure, personnel and services on children's nutritional status in rural Nepal. Data for the study come from the 1996 Nepal Living Standards Survey, which includes individual- and household-level information on children's nutritional status and its environmental and socioeconomic determinants, and community-level information on the availability of health care infrastructure, personnel and services. The study uses a structural modelling approach to assess the relative contributions of the health care supply environment on children's anthropometric status via the pathway of maternal and child health (MCH) service use. The findings suggest that improvements in the availability of outreach clinics and the structural quality of the closest public facility would be expected to have statistically significant and large effects on the use of MCH services, and that increases in MCH service use would have a statistically significant impact on weight-for-age, but not weight-for-height or height-for-age. The overall impact of the heath care supply environment on nutritional status is assessed through a series of policy simulations. PMID- 11926454 TI - Women's social power, child nutrition and poverty in Mali. AB - While the macro-level association between poverty and child malnutrition is well established, the concept of 'poverty' and its operationalization in terms of measures of socioeconomic status shed little or no light on the mechanisms through which malnutrition is created and/or prevented. This paper investigates a woman's social power, one such mechanism that may mediate the impact of poverty on childhood nutrition. This micro-level factor is examined using survey data on 402 children 5 years of age and younger and their 261 Fulbe mothers in rural Mali. A conceptual model of social power is developed and used to test the hypothesis that a mother's social power can predict her child's nutritional status. PMID- 11926455 TI - Reproductive health knowledge and use of services among young adults in Dakar, Senegal. AB - A study was conducted in Dakar, Senegal, to measure reproductive health knowledge and contraceptive use among young adults, and access to family planning services. A household survey was conducted with 1973 single and married women aged 15-24 and 936 single men aged 15-19. Two focus groups and a simulated client study were also conducted. The survey and focus groups noted gaps in knowledge of family planning methods and reproductive health. There were misconceptions about methods and only one-third of men and women aged 15-19 correctly identified the time of the menstrual cycle when a women is most likely to get pregnant. Contraceptive use at time of first premarital sexual experience was less than 30%. The simulated client study noted many barriers to services. 'Clients' felt uncomfortable in the clinics and felt that providers were reluctant to take care of them. None of the 'clients' who requested contraception received it. PMID- 11926457 TI - Birth seasonality in Jujeno (north-west Argentina) altitude populations. AB - Seasonality of births has been related to multiple factors, including environmental, biological and sociocultural ones, in populations of diverse geographic and cultural origin. However, the relationship between this variable of bioanthropologic interest and geographic altitude has not been adequately explored. The aim of this study was to analyse the monthly distribution pattern of birth rates in Jujeno (north-west Argentina) populations located at different geographic altitude levels. Data were provided by the Statistical Reports on Liveborns (n = 110,404) in the province of Jujuy, 1985-1992. Records were grouped by birthplace into the four geographic regions of the province: Ramal (350 m.a.s.l.), Valle (1200 m.a.s.l.), Quebrada (2500 m.a.s.l.) and Puna (3500 m.a.s.l.). Henry's coefficients of seasonality were estimated. A chi-square test for goodness of fit (chi2) was used to identify statistically significant seasonal variations. Edwards' test was used to detect simple harmonic cycles in birth distribution. Statistically significant seasonal patterns were found in the four geographic regions and they respond to a cyclic model of simple harmonic variation. The greatest Henry coefficients were observed in spring and summer in the higher regions (Puna and Quebrada). whereas they were seen in autumn and winter in the lowlands (Valle and Ramal). It is suggested that these patterns reflect the influence of environmental and socioeconomic altitude-related factors and inter-regional cultural diversity, rather than the influence of geographical altitude per se. PMID- 11926456 TI - Knowledge of possible pregnancy at first coitus: a study of in-school adolescents in Ibadan, Nigeria. AB - This paper discusses the reproductive health knowledge of Nigerian in-school adolescents, with special reference to pregnancy occurrence at first coitus. The data were derived from an Association for Reproductive and Family Health (ARFH) survey carried out in four secondary schools in Ibadan, Nigeria, between August and October 1995. A total of 828 students were interviewed. The results revealed that the majority of sexually active adolescents were not aware of the consequences of their actions. Religious affiliation and number of wives in a male adolescent's family, and religion and marital status of a female adolescent's parents, were found to influence adolescents' knowledge of pregnancy probability at first coitus. This paper confirms earlier findings that the majority of Nigerian adolescents do not know the consequences of sex. Therefore, a clear-cut plan of action is needed to inform sexually active adolescents through an effective sex education programme. PMID- 11926458 TI - Does education mediate the relationship between IQ and age of first birth? A behavioural genetic analysis. AB - This study presents a multivariate behavioural genetic analysis of the relationship between education, intelligence and age of first birth. Analyses investigated the mediational role of education in explaining the relationship between intelligence and age of first birth at both the phenotypic and behavioural genetic level. The data come from the National Longitudinal Survey of Youth (NLSY), a nationally representative survey that included genetically informative full- and half-sibling pairs (n = 1423 pairs). Respondents were aged 14 to 22 when contacted in 1979. Heritability estimates were 0.32, 0.50 and 0.06 for IQ, education and age of first birth, respectively. Shared environment estimates were 0.35, 0.23 and 0.20 respectively. Common genetic and shared environmental factors were substantial in explaining the relationship between intelligence and education, and also education and age of first birth. Education partially mediated the relationship between intelligence and age of first birth only in the phenotypic analyses. After considering the genetic and shared environmental factors that influence all three variables, evidence for mediation was less convincing. This pattern of results suggests that the apparent mediational role of education at the phenotypic level is in fact the result of underlying genetic and shared environmental influences that affect education, IQ and age of first birth in common. PMID- 11926459 TI - Social and lifestyle factors associated with diabetes in the adult Bahraini population. AB - This paper aimed to assess the prevalence of known diabetes among Bahraini adults, and to determine associated social and lifestyle factors. A community based survey was carried out on 514 adults aged 30-79 years. The overall prevalence of known diabetes was 9%. Using multivariate analysis, the risk of diabetes was found to be higher among older (50-79 years), female, illiterate, currently married, non-smoking people, those who did not walk regularly, overweight and obese people (BMI > or = 25), those who had a history of hypertension and those who consumed fresh vegetables more than 3 times a week. However, only obesity was found to be significantly associated with diabetes (OR = 1.83, CI 1.48-4.15). PMID- 11926460 TI - Health-related quality of life in patients with skull base tumours. AB - The objective of the investigation was to report on the health-related quality of life (HRQoL) of patients diagnosed with skull base tumours using the Short Form Health Survey questionnaire (SF-36). Those patients suffering with vestibular schwannoma were examined to determine the effect facial nerve function had on their quality of life. It took place at the tertiary referral centre at the Department of Clinical Neurosciences, Western General Hospital, Edinburgh. A prospective study of 70 consecutive patients was taken, who harboured the following tumours: 54 vestibular schwannomas, 13 meningiomas, two haemangioblastomas and one hypoglossal schwannoma. Patients were interviewed using the short form 36 (SF-36) questionnaire. Facial nerve function was assessed in those patients who had vestibular schwannomas. The entire cohort of live skull base patients were assessed after a median follow-up time of 38.4 months. Patients with vestibular schwannoma treated conservatively with interval MRI had a quality of life similar to t he normal population. Those who underwent surgery had a significant difference in two of the SF-36 domains. No statistically significant correlation was found at final assessment between the degree of facial nerve functioning and any of the domains of SF-36. Patients with non vestibular tumours had an impaired HRQoL in seven of the eight domains. Patients with skull base tumours have a significant impairment of their HRQoL. A conservative policy of follow up with interval MRI for patients with small vestibular schwannomas may therefore be more appropriate to preserve their HRQoL. Facial nerve outcome has little influence on quality of life in vestibular schwannoma patients. PMID- 11926461 TI - Demonstration of the optic pathway in large pituitary adenoma on heavily T2 weighted MR images. AB - The objective of the investigation was to understand preoperatively the detailed anatomical relationship of large pituitary adenomas to surrounding structures, using the heavily T2-weighted reversed (T2R) MR images. This study consisted of 28 patients with pituitary adenoma, presenting with visual disturbance. The MRI scanner used was a Gyroscan ACSNT 1.5T and the slice thickness of the image was 3 mm with 0.5 mm interslice gap. The relation of pituitary adenoma to optic pathway and to the degree of visual field defect was assessed. Relations of the optic chiasm to adenoma were classified into three types: anterior, superior and posterior. The optic chiasm was directly visualized and identifiable in all patients studied. It was located anterior in four cases, superior in 22 and posterior in two in relation to the adenoma. Its location was further confirmed by the anatomical delineation of surrounding structures such as anterior commissure and lamina terminalis. Optic nerve or tract was unidentifiable in one case, for each category. Detectability of each optic component was higher on T2R images than on conventional T1-weighted images. The adenoma extended into and in front of the third ventricle in anterior and posterior types, respectively. The anterior communicating artery complex and the optic pathway were relocated together in anterior and superior types, and were separated in the posterior type. In a case of the posterior type, the complex was sectioned to obtain a wider surgical field during anterior interhemispheric approach. While degrees of visual field defect were proportional to tumour size in the superior type, they were unrelated in the anterior and posterior types. On choosing a transcranial approach, the transcallosal route is unsuitable for an adenoma of posterior type, which extends in front of the third ventricle. This preoperative MRI information makes it possible to visualize directly the optic pathway even in huge adenomas, and is useful in predicting surgical anatomy and selecting a proper surgical approach. PMID- 11926462 TI - A laboratory model of testing shunt performance after implantation. AB - Constant rate infusion tests are used clinically to test shunt function in vivo in hydrocephalic patients. The criteria for appropriate shunt function have never been validated in the laboratory. Nine of the most commonly used types of hydrocephalus valves construction were selected and tested in a model of the CSF circulation incorporating increased resistance to CSF outflow [24 mmHg/(ml/min)] and decreased hydrodynamic compliance (<2 ml/mmHg), that are typical conditions in hydrocephalus. The aim was to document the pressure response to constant rate infusion of a model of CSF circulation with different valves and to define which measures are useful in shunt testing in vivo. The pressure-course of simulated CSF pressure was established and proved to be equivalent to clinical results. The baseline CSF pressure failed to correlate with shunt operating pressure for medium pressure valves (R = 0.14, p > 0.05). End-equilibrium pressure recorded during infusion correlated strongly with the opening pressure (R = 0.94, p = 0.0001) and the shunt's resistance (R = 0.86, p = 0.0026). The infusion test is able to assess shunt function. End-equilibrium pressure recorded during the test has been confirmed to correlate with the shunt's performance. PMID- 11926463 TI - Integration of image transmission into a protocol for head injury management: a preliminary report. AB - Neurosurgical care is limited in many parts of the world to one or two hospitals serving a large geographic area. The quality of neurosurgical response to emergencies depends on the reliability and completeness of the information received from referral hospitals. The aim of this study is to show how application of guidelines for head injury management in an entire area can be usefully combined with transmission of images from the peripheral to the central hospital. From January 1998 to December 2000, 1665 CT examinations were sent via image transfer to the Neurosurgical Unit; 637 first examinations (47%) and 206 second examinations (70%) were related to acute trauma cases. Out of 637 first examinations, 150 patients were actually transferred to the Neurosurgery Unit (23%), whereas of 206 second examinations, only 10 patients were secondarily transferred (5%). In the absence of the outcomes of patients located outside the Neurosurgical Unit, we studied in detail these 10 patients. They are, in fact, the only way for us to partially measure the impact of our system. Only in a single case could the death be attributed to a delay in transferring the patient. We then studied the factors influencing the decision of patient transfer. Mean GCS was 11 both for transferred and non transferred cases. The mean age of all patients was 52 years (median 48, SD 20.5 years); mean age of non-transferred patients was 54 years and for transferred patients it was 41 years (p < 0.01). The same statistically significant difference concerning age applied to any type of pathology sent via image link. In conclusion our data show that it is feasible to co-ordinate in an entire area the treatment of head injured patients. Available systems for CT images link are reliable and mostly useful. Unnecessary transfers can be avoided and the neurosurgeons can evaluate the images of a number of patients who have always been treated outside our Units. This results in more work for the neurosurgeons on duty, but also in a better quality service for the whole area. The lack of follow-up for patients not admitted to Neurosurgery is the limitation on a quality assessment of the system. PMID- 11926464 TI - Coil embolization of giant serpentine aneurysms: report of two cases arising from the posterior cerebral artery. AB - The authors describe the presentation, investigation and successful treatment of two giant serpentine aneurysms with Guglielmi detachable coils. Both aneurysms selectively involved the P2 segment of the posterior cerebral artery. The authors describe an endovascular approach to these challenging lesions as an alternative/adjunct to conventional surgery. PMID- 11926465 TI - Reversible brain injury in a head-injured patient identified by magnetic resonance imaging and spectroscopy. AB - Serial MRI studies of a severely head-injured patient showed extensive diffusion abnormalities lasting for 40 days. These were related to initial perfusion abnormalities and improved in parallel with clinical status. Spectroscopy showed diffuse damage and pH changes. Together these data support a role for ischaemia in head injury. PMID- 11926466 TI - Microvascular decompression of a developmental venous anomaly in the cerebellopontine angle causing trigeminal neuralgia. AB - We describe an apparently unique case of a patient with a trigeminal neuralgia caused by compression of the trigeminal nerve during its course by the draining vein of a developmental venous anomaly in the cerebellopontine angle. Typical symptoms of trigeminal neuralgia disappeared completely after microvascular decompression of the nerve. Neuroradiological findings, as well as particularities of this case are described and therapeutic options are discussed. PMID- 11926467 TI - Neurosurgical treatment of short-lasting, unilateral, neuralgiform hemicrania with conjunctival injection and tearing. AB - Trigeminal neuralgia solely involving the upper trigeminal nerve branch is rare. The SUNCT syndrome (short-lasting, unilateral, neuralgiform hemicrania with conjunctival injection and tearing) in which the periorbital pain lasts for 60 120 s, and is accompanied by conjunctival injection and tearing is even less common. Unlike trigeminal neuralgia, SUNCT is usually not relieved by medication. Three patients with SUNCT were treated with retrogasserian glycerol rhizolysis, two of them twice. All five treatments provided complete pain relief and the duration of the effects was 2 to more than 4 years. One of these three patients also had a third treatment with compression of retroganglionic fibres with a Fogarthy balloon, according to Mullan, of the upper trigeminal nerve with excellent results. PMID- 11926468 TI - Heparin treatment of atrial thrombus in ventriculo-atrial shunts. AB - This short report details the management of two cases of atrial thrombus on ventriculoatrial shunts, treated non-surgically by simple anticoagulation. While this is extremely simple it appears not to have been reported before. PMID- 11926469 TI - Intraparenchymal haemorrhage after evacuation of chronic subdural haematoma. Report of three cases and review of the literature. AB - Three cases of intracerebral haemorrhage following removal of a chronic subdural haematoma are reported and the literature on this topic reviewed. The possibility of an increase in cerebral blood flow following removal of CSH, makes slow, gradual decompression mandatory in all patients submitted to cranial trapanation. PMID- 11926470 TI - Metastasis of pulmonary adenocarcinoma in right sylvian secretory meningioma. AB - We report a secretory meningioma with metastasis from a pulmonary adenocarcinoma in a 48-year-old woman. Secretory meningioma can simulate metastatic disease both clinically and pathologically. Secretory meningioma and tumour-to-tumour metastasis are each rare, and we believe this to be the first report of their coincidence. PMID- 11926472 TI - Cystic meningioma: challenges in the diagnosis. AB - A patient with an unusual cystic meningioma is described. The pathophysiology of cyst formation is discussed. PMID- 11926471 TI - Osteoid osteomas of the body of the cervical spine. Case report and review of the literature. AB - A case report of an adult patient with an osteoid osteoma of the body of the 3rd cervical vertebra is presented. The long history of progressively worsening nocturnal cervical pain which was relieved by non-steroidal anti-inflammatory drugs, and an essentially normal clinical examination, are typical of this condition; in which CT is the investigation of choice. Complete relief was obtained following excision of the lesion. The literature is reviewed. PMID- 11926473 TI - Frontal lobe tuberculoma masquerading as postnatal depression. PMID- 11926474 TI - Intramedullary spinal tuberculoma. PMID- 11926475 TI - Traumatic tension pneumocephalus presenting with blindness. PMID- 11926476 TI - Intraventricular cavernoma: unusual occurrence in the region of the foramen of monro. PMID- 11926477 TI - Neurocysticercosis in UK. PMID- 11926478 TI - A descriptive analysis of research methods classes in departments of kinesiology and physical education in the United States. AB - Research training takes many forms and is generally a part of graduate education. A common and important aspect of research training is the introductory research methods class offered by many departments. The purpose of this study was to examine the content, process, and instructors of introductory research methods classes in departments of kinesiology and physical education in the United States. A survey was designed and extensively pilot tested. The sample was selected from all departments offering graduate degrees in the United States. Among the many results, the data indicate that one book was required reading in more than half the classes and class size averaged about 19 students. A number of objectives were statedfor most classes, with understanding research, applying research to professional situations, critiquing the research literature, and planning research indicated most often. Quantitative design and analysis topics were emphasized more strongly than qualitative design and analysis topics. Professors indicated that more than half the class time was spent lecturing and most grades were based on exams, preparation of a research proposal, and regular assignments. The professors were relatively experienced, had a variety ofspecialty areas, and were reasonably productive researchers. The trends suggest that alternative research methodologies have not been quickly added to the research methods curriculum. PMID- 11926479 TI - Arm movement control: differences between children with and without attention deficit hyperactivity disorder. AB - The purpose of this study was to examine performance differences in arm movement control (programmingvs. "on-line" control) between children with and without attention deficit hyperactivity disorder (ADHD). Twenty children (10 with ADHD and 10 without ADHD) from the ages of 8 to 13 years participated in the study. On the surface of a digitizer, each participant completed three types of aiming arm movements (10 trials for each) and 10 baseline trials (without accuracy requirement). Multivariate analyses of variance with repeated measures were used to analyze the variables of reaction time, movement time, normalized jerk, intersegment-interval (ISI), and movement timing. Children with ADHD appeared to use "on-line " monitoring during the arm movement and did not perform the entire movement sequence as afunctional unit. They executed the arm movements more slowly, had greater variability in movement timing, and demonstrated longer ISIs than their counterparts. Children with ADHD had multiple peaks in the velocity profiles. Children withoutADHD, however, appeared to program their entire arm movements and execute the sequence as a unit. Their velocity profiles were symmetrical with a single peak, and the movement segments were temporally coordinated. Thesefindings suggested that cognitive functions are important resources for controlling rapid aiming arm movements. Children with ADHD might rely more on visual feedback during the movements, which resulted in slower and more variant movement outcomes than children who did not have ADHD. PMID- 11926480 TI - Visual search and biological motion perception in tennis. PMID- 11926481 TI - Physical self-concept and strength changes in college weight training classes. PMID- 11926482 TI - Throwing patterns used by collegiate baseball players in actual games. AB - The form of 3,684 throws made by 100 players in 7 collegiate baseball games was examined in relation to position, distance, and active and inactive situations. All throws made from the first pitch to the last out for 94 half innings were videotaped from the stands. The results showed that the interrater reliability of task, environment, and performance measures were all acceptable to excellent (percentage of agreement was at least 80%), indicating that qualitative aspects of throwing can be reliably measured in an actual sport context. Alternatives to the most advanced pattern were observed at least half the time for catchers and infielders at most distances in both active and inactive situations andfor pitchers and outfielders only in inactive situations. PMID- 11926483 TI - Running, heart disease, and the ironic death of Jim Fixx. AB - Jim Fixx was one of millions ofAmericans who started running in the 1 960s, 1970s, and 1980s. Unlike other runners, however, Fixx wrote a best-selling book about running and, ironically, died of a heart attack at the age of 52 years while running. Fixx and the authors of other running books believed heart disease resulted from overcivilization and recommended running as a cure. Running was not merely a physical exercise, according to those authors, but also a way of life. Moreover, those running authors, who were often doctors themselves, advised their readers to listen to their bodies, instead of their doctors. Fixx's adherence to that philosophy offers an explanationfor his seemingly irrational behavior- running through chest pain and discomfort. PMID- 11926484 TI - The role of parameter variability on retention, parameter transfer, and effector transfer. AB - The purpose of this study was to examine the effects of parameter variability on the learning of generalized motor programs (GMP) and movement parameterization. Participants attempted to exert a force pattern that resembled in force and time a waveform displayed on a computer monitor. The analysis suggested that relative timing (a measure of the GMP) performance remained remarkably stable across retention and transfer tests, whereby the structure of the movement remained intact, although the parameter or muscle group (effector) changed during transfer The results also indicated that variable parameter (time) practice did not enhance GMP learning but did degrade the learning parameter that was not varied (force). In addition, parameter specification was substantially less stable than the GMP, with time and force parameter performance deteriorating from the retention to transfer tests. These findings suggest that parameter specification, and not the GMP, is the primary cause of poorer performance in parameter and effector transfer. PMID- 11926485 TI - Achievement goals, perceived motivational climate, and students' self-reported mastery behaviors. AB - Achievement goals and perceived motivational climate are two important constructs in achievement goal theory, and they play critical roles in student motivation and behavior Traditionally, these two constructs have been examined separately. The present study examined relationships between the two constructs and students' self-reported mastery behaviors as well as age-related differences. Three hundred eight students in 4th, 8th, and 11th grades completed questionnaires assessing their achievement goals, perceived motivational climate, and perceptions of their mastery behaviors. Results indicated that achievement goals and perceived motivational climate were related to students' self-reported mastery behaviors. However, the relations varied depending on the students' ages. The older students, compared to the younger ones, appeared to be inclined more toward ego orientation and the ego-involved climate. A multiple achievement goals perspective is recommended for future research on relationships among achievement goals, perceived motivational climate and achievement-related cognitions, and behaviors in physical education. PMID- 11926486 TI - Prediction of maximum oxygen consumption from walking, jogging, or running. AB - The purpose of this study was to develop a submaximal, 1.5-mile endurance test for college-aged students using walking, jogging, or running exercise. College students (N = 101: 52 men, 47 women), ages 18-26years, successfully completed the 1.5-mile test twice, and a maximal graded exercise test. Participants were instructed to achieve a "somewhat hard" exercise intensity (rating of perceived exertion = 13) and maintain a steady pace throughout each 1.5-mile test. Multiple linear regression generated the following prediction equation: VO2 max = 65.404 + 7.707 x gender (1 = male; 0 =female) - 0.159 x body mass (kg) - 0.843 x elapsed exercise time (min; walking, jogging orrunning). This equation shows acceptable validity (R = .86, SEE = 3.37 ml x kg(-1) min(-1)) similar to the accuracy of comparable field tests, and reliability (ICC = .93) is also comparable to similar models. The statistical shrinkage is minimal (R(press) = 0.85, SEE(press) = 3.51 ml x kg(-) x min(-1)); hence, it should provide comparable results when applied to other similar samples. A regression model (R =.90, and SEE = 2.87 ml x kg(-1) min(-1)) including exercise heart rate was also developed: VO2 max = 100.162 +/- 7.301 x gender(1 = male; 0 =female) - 0.164 x body mass (kg) - 1.273 x elapsed exercise time -0.156 x exercise heart rate, for those who have access to electronic heart rate monitors. This submaximal 1.5-mile test accurately predicts maximal oxygen uptake (VO2max) without measuring heart rate and is similar to the 1.5-mile run in that it allowsfor mass testing and requires only a flat, measured distance and a stopwatch. Further, it can accommodate a wide range of fitness levels (from walkers to runners). PMID- 11926487 TI - Coaching commitment and turnover: a comparison of current and former coaches. AB - Although coaching continuity is thought to be important in developing and sustaining quality sport programs, coach turnover is not well understood. Consequently, we used a commitment model to address why some coaches maintain their involvement while others discontinue coaching. For this study, 469 USA Swimming age-group coaches completed a mail survey designed to assess commitment model constructs, including coaching satisfaction, benefits, costs, investments, alternative options, social constraints, and commitment. To test the commitment model, we first used structural equation modeling to assess whether the model provided an adequate fit to the data for current and former coaches. Overall, satisfaction and investments were significantly related to commitment. Moreover, the standardized path coefficients between benefits and costs and satisfaction were also significant. Multivariate analysis of variance andfollow-up analyses revealed that current coaches reported higher commitment as well as less attractive alternative options, higher investments, and higher social constraints compared to former coaches. In replication of past research, we compared current andformer coaches on the benefits and costs associated with coaching. PMID- 11926488 TI - Perceived control of anxiety and its relationship to self-confidence and performance. AB - This study examined performers' retrospective explanations of the relationship between anxiety symptoms, self-confidence, and performance. Interviews were used to determine how the presence of symptoms and the accompanying directional interpretation affected performance in six elite and six subelite swimmers. Causal networks revealed that perceived control was the moderatingfactor in the directional interpretation of anxiety and not the experience of anxiety symptoms alone. Symptoms perceived to be under control were interpreted to have facilitative consequences for performance; however, symptoms not under control were viewed as debilitative. Increases or decreases in self-confidence wereperceived to improve or lower performance. Findings reveal how cognitive and somatic information was processed, what strategies were adopted, and how this series of events related to performance. PMID- 11926489 TI - EMG power frequency spectrum shifts during repeated isokinetic knee and arm movements. PMID- 11926490 TI - Within-host population dynamics and the evolution of microparasites in a heterogeneous host population. AB - Why do parasites harm their hosts? The general understanding is that if the transmission rate and virulence of a parasite are linked, then the parasite must harm its host to maximize its transmission. The exact nature of such trade-offs remains largely unclear, but for vertebrate hosts it probably involves interactions between a microparasite and the host immune system. Previous results have suggested that in a homogeneous host population in the absence of super- or coinfection, within-host dynamics lead to selection of the parasite with an intermediate growth rate that is just being controlled by the immune system before it kills the host (Antia et al. 1994). In this paper, we examine how this result changes when heterogeneity is introduced to the host population. We incorporate the simplest form of heterogeneity--random heterogeneity in the parameters describing the size of the initial parasite inoculum, the immune response of the host, and the lethal density at which the parasite kills the host. We find that the general conclusion of the previous model holds: parasites evolve some intermediate growth rate. However, in contrast with the generally accepted view, we find that virulence (measured by the case mortality or the rate of parasite-induced host mortality) increases with heterogeneity. Finally, we link the within-host and between-host dynamics of parasites. We show how the parameters for epidemiological spread of the disease can be estimated from the within-host dynamics, and in doing so examine the way in which trade-offs between these epidemiological parameters arise as a consequence of the interaction of the parasite and the immune response of the host. PMID- 11926491 TI - The evolution of the vertebrate beta-globin gene promoter. AB - Complexity analysis is capable of highlighting those gross evolutionary changes in gene promoter regions (loosely termed "promoter shuffling") that are undetectable by conventional DNA sequence alignment. Complexity analysis was therefore used here to identify the modular components (blocks) of the orthologous beta-globin gene promoter sequences of 22 vertebrate species, from zebrafish to humans. Considerable variation between the beta-globin gene promoters was apparent in terms of block presence/absence, copy number, and relative location. Some sequence blocks appear to be ubiquitous, whereas others are restricted to a specific taxon. Block similarities were also evident between the promoters of the paralogous human beta-like globin genes. It may be inferred that a wide variety of different mutational mechanisms have operated upon the beta-globin gene promoter over evolutionary time. Because these include gross changes such as deletion, duplication, amplification, elongation, contraction, and fusion, as well as the steady accumulation of single base-pair substitutions, it is clear that some redefinition of the term "promoter shuffling" is required. This notwithstanding, and as previously described for the vertebrate growth hormone gene promoter, the modular structure of the beta-globin promoter region and those of its paralogous counterparts have continually been rearranged into new combinations through the alteration, or shuffling, of preexisting blocks. Some of these changes may have had no influence on promoter function, but others could have altered either the level of gene expression or the responsiveness of the promoter to external stimuli. The comparative study of vertebrate beta-globin gene promoter regions described here confirms the generality of the phenomenon of sequence block shuffling and thus supports the view that it could have played an important role in the evolution of differential gene expression. PMID- 11926492 TI - Phylogeny, concerted convergence, and phylogenetic niche conservatism in the core Liliales: insights from rbcL and ndhF sequence data. AB - Calochortus and the family Liliaceae s.s. have often been considered each other's closest relatives, based partly on their shared possession of bulbs, visually showy flowers, winged wind-dispersed seeds, and narrow parallel-veined leaves. We present a well-supported molecular phylogeny for these groups and their close relatives in the core Liliales, based on sequence variation in the chloroplast encoded rbcL and ndhF genes. This analysis identifies Liliaceae s.s. as monophyletic. including one clade (((Lilium, Fritillaris, Nomocharis), Cardiocrinum), Notholirion) that appears to have diversified in the Himalayas roughly 12 million years ago and another ((Erythronium, Tulipa), (Gagea, Lloydia)) that arose in East Asia at about the same time. Medeola and Clintonia are sister to Liliaceae s.s. and bear rhizomes, inconspicuous flowers, fleshy animal-dispersed fruits, and broad reticulate-veined leaves. Calochortus is sister to Tricyrtis; both Tricyrtis and the neighboring clade of Prosartes Streptopus-Scoliopus share several of the traits seen in Medeola-Clintonia. The core Liliales thus provide compelling examples of both concerted convergence and phylogenetic niche conservatism. Invasion of open, seasonal habitats was accompanied by the independent evolution of bulbs, showy flowers, wind-dispersed seeds, and narrow parallel-veined leaves in Calochortus and Liliaceae s.s. Conversely, persistence in shady habitats was accompanied by the retention of rhizomes, inconspicuous flowers, animal-dispersed seeds, and broad reticulate veined leaves in their sister groups. We advance arguments for the context specific adaptive value of each of these traits, as well as evidence of parallel trends in other groups. Concerted convergence--convergence in several different traits, favored by the same shared set of ecological conditions, in two or more lineages--is an important evolutionary process that can mislead evolutionary analyses based solely on phenotypic variation. PMID- 11926493 TI - Mitochondrial DNA diversity, population structure, and gender association in the gynodioecious plant Silene vulgaris. AB - A highly variable mitochondrial DNA (mtDNA) restriction fragment length polymorphism (RFLP) locus is used to assess the population structure of mitochondrial genomes in the gynodioecious plant Silene vulgaris at two spatial scales. Thirteen mtDNA haplotypes were identified within 250 individuals from 18 populations in a 20-km diameter region of western Virginia. The population structure of these mtDNA haplotypes was estimated as thetaST = 0.574 (+/- 0.066 SE) and, surprisingly, genetic differentiation among populations was negatively correlated with geographic distance (Mantel r = -0.246, P < 0.002). Additionally, mtDNA haplotypes were spatially clumped at the scale of meters within one population. Gender in S. vulgaris is determined by an interaction between autosomal male fertility restorers and cytoplasmic male sterility (CMS) factors, and seed fitness is affected by an interaction between gender and population sex ratio; thus, selection acting on gender could influence the distribution of mtDNA RFLP haplotypes. The sex ratio (females:hermaphrodites) varied among mtDNA haplotypes across the entire metapopulation, possibly because the haplotypes were in linkage disequilibrium with different CMS factors. The gender associated with some of the most common haplotypes varied among populations, suggesting that there is also population structure in male fertility restorer genes. In comparison with reports of mtDNA variation from other published studies, we found that S. vulgaris exhibits a large number of mtDNA haplotypes relative to that observed in other species. PMID- 11926494 TI - The inheritance of modifiers conferring self-fertility in the partially self incompatible perennial, Campanula rapunculoides L. (Campanulaceae). AB - The role of partial self-incompatibility in plant breeding system evolution has received little attention. Here, we examine the genetic basis of modifiers conferring self-fertility in the creeping bellflower, Campanula rapunculoides L. (Campanulaceae), a partially self-incompatible herb. A survey of 35 individuals from two natural populations indicates that 45% of them are strongly self incompatible, 40% intermediately self-incompatible, and 15% weakly self incompatible and that some plants show a strong breakdown in self-incompatibility over floral age. We generated 101 F1 families by random crossing among 31 parental plants and estimated the heritability of self-fertility in day 1 and day 4 female-phase flowers, the genetic correlation between day 1 and day 4 self fertility, and the coefficient of additive genetic variance of self-fertility. We use linear regression and data from additional crosses to examine whether there are significant maternal effects in the expression of self-fertility. We use Fain's test to determine if a major gene influences self-fertility and, finding no evidence, use data from additional crosses on an F2 generation to estimate the mean number and dominance of genes conferring self-fertility. These analyses indicate that the heritability (h2) of self-fertility is 0.24 in day 1 female phase flowers and 0.44 in day 4 flowers, self-fertility is primarily additive but shows some recessive effects, and self-fertility is estimated to be controlled by four genetic factors. In addition, we have evidence that there may be maternal effects for self-fertility, especially for weakly self-incompatible plants. The significance of these results in the context of mating system evolution is discussed. PMID- 11926495 TI - Morphological traits defining species differences in wild relatives of maize are controlled by multiple quantitative trait loci. AB - We analyzed the genetic basis of morphological differences between two wild species of teosinte (Zea diploperennis and Z. mays ssp. parviglumis), which are relatives of maize. These two species differ in a number of taxonomically important traits including the structure of the tassel (male inflorescence), which is the focus of this report. To investigate the genetic inheritance of six tassel traits, quantitative trait locus (QTL) mapping with 95 RFLP markers was employed on a population of 425 F2 plants. Each trait was analyzed by interval mapping (IM) and composite interval mapping (CIM) to identify and characterize the QTL controlling the differences in tassel morphology. We detected two to eight QTL for each trait. In total, 30 QTL with IM and 33 QTL with CIM were found for tassel morphology. QTL for several of the traits mapped near each other, suggesting pleiotropy and/or linkage of QTL. The QTL showed small to moderate magnitudes of effect. No QTL of exceptionally large effect were found as seen under domestication and in the case of some other natural species. Thus, the model involving major QTL of large effect seems not to apply to the traits and species analyzed. A mixture of QTL with positive and negative allelic effects was found for most tassel traits and may suggest a history of periodic changes in the direction of selection during the divergence of Z. diploperennis and Z. mays ssp. parviglumis or fixation of QTL alleles by random genetic drift rather than selection. PMID- 11926496 TI - Escalation and extinction selectivity: morphology versus isotopic reconstruction of bivalve metabolism. AB - Studies that have tested and failed to support the hypothesis that escalated species (e.g., those with predation-resistant adaptations) are more susceptible to elimination during mass extinctions have concentrated on the distribution and degree of morphological defenses in molluscan species. This morphological approach to determining level of escalation in bivalves may be oversimplified because it does not account for metabolic rate, which is an important measure of escalation that is less readily accessible for fossils. Shell growth rates in living bivalves are positively correlated with metabolic rate and thus are potential indicators of level of escalation. To evaluate this approach, we used oxygen isotopes to reconstruct shell growth rates for two bivalve species (Macrocallista marylandica and Glossus markoei) from Miocene-aged sediments of Maryland. Although both species are classified as non-escalated based on morphology, the isotopic data indicate that M. marylandica was a faster-growing species with a higher metabolic rate and G. markoei was a slower-growing species with a lower metabolic rate. Based on these results, we predict that some morphologically non-escalated species in previous tests of extinction selectivity should be reclassified as escalated because of their fast shell growth rates (i.e., high metabolic rates). Studies that evaluate the level of escalation of a fauna should take into account the energetic physiology of taxa to avoid misleading results. PMID- 11926497 TI - Assortative fertilization and selection at larval stage in the mussels Mytilus edulis and M. galloprovincialis. AB - Assortative mating (prezygotic isolation) and reduced hybrid fitness (postzygotic isolation) are typically invoked to explain the stability of hybrid zones. In the tension zone model, these factors work in opposition to migration, which promotes genetic homogeneity. Many marine animals migrate over long distances through a planktonic larval stage. Therefore, strong reproductive isolation is needed to maintain stable marine hybrid zones. However, surprisingly little is known about mating preferences and hybrid fitness in marine organisms. Smooth-shelled mussels (Mytilus spp.) form a well-known species complex, with hybridization over extensive areas such as the contact zone of M. edulis and M. galloprovincialis around European Atlantic coasts. This paper reports direct experimental evidence of assortative fertilization, hybrid larval inviability, and early heterosis for growth rate in M. edulis and M. galloprovincialis. Four crosses between pure M. edulis and M. galloprovincialis were analyzed with a new polymerase-chain reaction-based diagnostic marker. Gamete competition between taxa was allowed in two out of the four crosses. Genotype frequencies observed at an early stage (36 h after fertilization) unambiguously revealed assortative fertilization when gamete competition was allowed. A significant reduction in hybrid viability was subsequently observed during the larval stage. At the same stage an antagonistic effect, heterosis, was observed on growth rate. However, even if heterosis is observed in the F1, it is expected to vanish in subsequent hybrid generations. Although specialization for different habitats and asynchronous spawning have been mentioned as factors contributing to the maintenance of the blue mussel hybrid zone in Europe, we argue that assortative fertilization and reduced hybrid fitness are important factors that also contribute to the stabilization of this zone. These results emphasize that multiple factors may act concomitantly in a barrier to gene flow, especially in complex life cycles. Furthermore, they show that assortative mating through gamete preference, as already demonstrated for sea urchins, may play a role in speciation processes taking place in the sea. PMID- 11926498 TI - Temperature-induced shifts in associations of longevity with body size in Drosophila melanogaster. AB - One of the hypotheses of growing interest in studies of responses to thermal environments suggests that trade-offs and other trait associations may be altered by temperature. Here, the commonly observed positive association between body size and longevity was examined at two adult test temperatures, 14 degrees C and 25 degrees C, in cold-stress-selected lines (S) and their controls (C) in 25 degrees C-reared Drosophila melanogaster. Thorax length (TL) and developmental time (DT) were also scored in 25 degrees C-reared individuals before and after one generation of truncation selection on longevity. The topography of the selection surface that relates longevity to thorax and wing size was temperature dependent and differed both between lines and between sexes. Longevity increased monotonically with body size (TL) in C and S females at 25 degrees C but, surprisingly, longevity decreased with body size in S individuals at 14 degees C. Body size did not diverge between S and C lines and showed no response to longevity selection. However, DT increased by 25 degrees C-longevity selection in C individuals and decreased by 14 degrees C-longevity selection in S individuals. These results suggest that trait associations (including the commonly observed trade-off between body size and DT) can greatly depend on temperature, as a shift in the sign of the correlation is possible at low temperature. Genotype x temperature interaction is an important source of variation in the relationship between soma size and longevity. PMID- 11926499 TI - Population variation in sexual selection and its effect on size allometry in two dung fly species with contrasting sexual size dimorphism. AB - Body size is one of the most important quantitative traits under evolutionary scrutiny. Sexual size dimorphism (SSD) in a given species is expected to result if opposing selection forces equilibrate differently in both sexes. We document variation in the intensity of sexual and fecundity selection, male and female body size, and thus SSD among 31 and 27 populations of the two dung fly species, Scathophaga stercoraria and Sepsis cynipsea, across Switzerland. Whereas in S. cynipsea females are larger, the SSD is reversed in S. stercoraria. We comprehensively evaluated Fairbairn and Preziosi's (1994) general, three-tiered scenario, hypothesizing that sexual selection for large male size is the major driving force of SSD allometry within these two species. Sexual selection intensity on male size in the yellow dung fly, S. stercoraria, was overall positive, greater, and more variable among populations than fecundity selection on females. Also, sexual selection intensity in a given population correlated positively with mean male body size of that population for both the field-caught fathers and their laboratory-reared sons, indicating a response to selection. In S. cvnipsea, sexual selection intensity on males was lower overall and significantly positive, about equal in magnitude, but more variable than fecundity selection on females. However, there was no correlation between the intensity of sexual selection and mean male body size among populations. In both species, the laboratory-reared offspring indicate genetic differentiation among populations in body size. Despite fulfillment of all key prerequisites, at least in S. stercoraria, we did not find hypoallometry for SSD (Rensch's rule, i.e., greater evolutionary divergence in male size than female size) for the field caught parents or the laboratory-reared offspring: Female size was isometric to male size in both species. We conclude that S. cynipsea does not fit some major requirements of Fairbairn and Preziosi's (1994) scenario, whereas for S. stercoraria we found partial support for it. Failure to support Rensch's rule within the latter species may be due to phylogenetic or other constraints, power limitations, erroneous estimates of sexual selection, insufficient genetic isolation of populations, or sex differences in viability selection against large size. PMID- 11926500 TI - A test of ecologically dependent postmating isolation between sympatric sticklebacks. AB - Ecological speciation occurs when reproductive isolation evolves ultimately as a result of divergent natural selection between populations inhabiting different environments or exploiting alternative resources. I tested a prediction of the ecological model concerning the fitness of hybrids between two young, sympatric species of threespine sticklebacks (Benthics and Limnetics). The two species are ecologically and morphologically divergent: the Benthic is adapted to feeding on invertebrates in the littoral zone of the lake whereas the Limnetic is adapted to feeding on zooplankton in the open water. The growth rate of two types of hybrids, the Benthic backcross and the Limnetic backcross, as well as both parent species, was evaluated in enclosures in both parental habitats in the lake. The use of backcrosses is ideal because a comparison of their growth rates in the two habitats estimates an ecologically dependent component of their fitness while controlling for any intrinsic genetic incompatibilities that may exist between the Benthic and Limnetic genomes. The backcross results revealed a striking pattern of ecological dependence: in the littoral zone, Benthic backcrosses grew at approximately twice the rate of Limnetic backcrosses, while in the open water, Limnetic backcrosses grew at approximately twice the rate of Benthic backcrosses. Such a reversal of relative fitness of the two cross-types in the two environments provides strong evidence that divergent natural selection has played a central role in the evolution of postmating isolation between Benthics and Limnetics. Although the rank order of growth rates of all cross-types in the littoral zone was Benthic > Benthic backcross > Limnetic backcross > Limnetic, neither backcross differed significantly from the parent from which it was mainly derived. Implications of this result are discussed in terms of ecological speciation and possible introgressive hybridization between the species. Results in the open water were less clear and were not fully consistent with the ecological model of speciation, mainly as a result of the low growth rate of Limnetics. However, analysis of the diet of the fish in the open water suggests that these enclosures may not have been fully successful at replicating the food regimes characteristic of this habitat. PMID- 11926501 TI - Phylogeography of the socially polymorphic sweat bee Halictus rubicundus (Hymenoptera: Halictidae). AB - The evolution of sociality in insects holds a central place in evolutionary theory. By examining the phylogenetic patterns of solitary and social behavior and how they correlate with ecological variables, we may identify factors important in the evolution of sociality. In this study, we investigated historical and biogeographical patterns of sociality in a socially polymorphic bee species (one that demonstrates both social and solitary nesting behavior). This unique system allows for a more powerful examination of evolutionary transitions in sociality than interspecific studies of obligately social and solitary species. We conducted a phylogenetic analysis among populations of the halictine bee Halictus rubicundus and then identified relationships among mitochondrial DNA sequence data, sociality, environmental conditions at the nesting site, and geographic location of populations of this species. Within North America, populations of H. rubicundus expressing social and solitary behavior belong to different genetic lineages. Sociality is also correlated with at least one environmental variable used in this study. Taken together, the results support the predictions for genetic control of sociality, but they are still consistent with social behavior at some level being determined by the environmental conditions at the nesting site. PMID- 11926502 TI - The environmental and genetic control of seasonal polyphenism in larval color and its adaptive significance in a swallowtail butterfly. AB - Seasonal polyphenism, in which different forms of a species are produced at different times of the year, is a common form of phenotypic plasticity among insects. Here I show that the production of dark fifth-instar caterpillars of the eastern black swallowtail butterfly, Papilio polyxenes, is a seasonal polyphenism, with larvae reared on autumnal conditions being significantly darker than larvae reared on midsummer conditions. Both rearing photoperiod and temperature were found to have individual and synergistic effects on larval darkness. Genetic analysis of variation among full-sibling families reared on combinations of two different temperatures and photoperiods is consistent with the hypothesis that variation in darkness is heritable. In addition, the genetic correlation in larval darkness across midsummer and autumnal environments is not different from zero, suggesting that differential gene expression is responsible for the increase in larval darkness in the autumn. The relatively dark autumnal form was found to have a higher body temperature in sunlight than did the lighter midsummer form, and small differences in temperature were found to increase larval growth rate. These results suggest that this genetically based seasonal polyphenism in larval color has evolved in part to increase larval growth rates in the autumn. PMID- 11926503 TI - Coevolution of color pattern and thermoregulatory behavior in polymorphic pygmy grasshoppers Tetrix undulata. AB - Ectothermic organisms, such as insects and reptiles, rely on external heat sources to control body temperature and possess physiological and behavioral traits that are temperature dependent. It has therefore been hypothesised that differences in body temperature resulting from phenotypic properties, such as color pattern, may translate into selection against thermally inferior phenotypes. We tested for costs and benefits of pale versus dark coloration by comparing the behaviors (i.e., basking duration and bouts) of pygmy grasshopper (Tetrix undulata) individuals exposed to experimental situations imposing a trade off between temperature regulation and feeding. We used pairs consisting of two full-siblings of the same sex that represented different (genetically coded) color morphs but had shared identical conditions from the time of fertilization. Our results revealed significant differences in behavioral thermoregulation between dark and pale individuals in females, but not in males. Pale females spent more time feeding than dark females, regardless of whether feeding was associated with a risk of either hypothermia or overheating. In contrast, only minor differences in behavior (if any) were evident between individuals that belonged to the same color morph but had been painted black or gray to increase and decrease their heating rates. This suggests that the behavioral differences between individuals belonging to different color morphs are genetically determined, rather than simply reflecting a response to different heating rates. To test for effects of acclimation on behaviors, we used pairs of individuals that had been reared from hatchlings to adults under controlled conditions in either low or high temperature. The thermal regime experienced during rearing had little effect on behaviors during the experiments reported above, but significantly influenced the body temperatures selected in a laboratory thermal gradient. In females (but not in males) preferred body temperature also varied among individuals born to mothers belonging to different color morphs, suggesting that a genetic correlation exists between color pattern and temperature preferences. Collectively, these findings, at least in females, are consistent with the hypothesis of multiple-trait coevolution and suggest that the different color morphs represent alternative evolutionary strategies. PMID- 11926504 TI - Phenotypic lability and the evolution of predator-induced plasticity in tadpoles. AB - The hypothesis that predator-induced defenses in anuran larvae are maintained by divergent selection across multiple predation environments has not been fully supported by empirical results. One reason may be that traits that respond slowly to environmental variation experience a fitness cost not incorporated in the standard adaptive model, due to a time lag between detecting the state of the environment and expressing the phenotypic response. I measured the rate at which behavior and morphology of Rana temporaria tadpoles change when confronted with a switch in the predation environment at two points in development. Hatchling tadpoles that had been exposed during the egg stage to Aeshna dragonfly larvae were not phenotypically different from those exposed as eggs to predator-free conditions, and both responded similarly to post-hatching predator treatments. When 25-day-old tadpoles from treatments with and without dragonflies were subjected to a switch in the environment, their activity budgets reversed completely within 24-36 h, and their body and tail shape began changing significantly within 4 days. The behavioral response was conservative: Tadpoles switched from high-risk to predator-free treatments were slower to adjust their activity. The study confirmed that behavioral traits are relatively labile and exhibit strong plasticity, but it did not reveal such a pattern at the level of individual traits: Morphological traits that developed slowly did not show the least plasticity. Thus, I found that differences in lability of traits were useful for predicting the magnitude of plasticity only for fundamentally different kinds of characters. PMID- 11926505 TI - Mate choice, genetic incompatibility, and outbreeding in the ornate dragon lizard, Ctenophorus ornatus. AB - There has been recent interest in the role genetic incompatibility may play in mate or sperm choice. One source of incompatibility may be too similar or disparate genomes. An isolated population of the ornate dragon lizard, Ctenophorus ornatus, was followed over a breeding season and parentage assigned to the offspring using microsatellites. It was found that the adults in the population had an eight per cent chance of mating with a relative. I examined whether C. ornatus mate (or fertilize their eggs) with respect to genetic similarity. There was no difference in a female's relatedness to the male in whose territory she resided and her average relatedness to all other males. Neither was there a difference in the relatedness of the male that sired a female's offspring and the female's average relatedness to all other males. There was no evidence of a cost to mating with a more genetically similar mate, because offspring survival was not influenced by degree of inbreeding or outbreeding. However, females that were more outbred had fewer offspring survive. In this small population there are two possible explanations for the decreased fitness associated with outbreeding. First, the species may have an evolutionary history of inbreeding and thus may be susceptible to outbreeding depression. Second. as fitter individuals produce more offspring, these offspring have an increased probability of breeding with relatives, leading to an indirect relationship between fitness and outbreeding. PMID- 11926506 TI - Isolation by distance and vicariance drive genetic structure of a coral reef fish in the Pacific Ocean. AB - We studied the genetic diversity of a coral reef fish species to investigate the origin of the differentiation. A total of 727 Acanthurus triostegus collected from 15 locations throughout the Pacific were analyzed for 20 polymorphic loci. The genetic structure showed limited internal disequilibrium within each population; 3.7% of the loci showed significant Hardy-Weinberg disequilibrium, mostly associated with Adh*, and we subsequently removed this locus from further analysis of geographic pattern. The genetic structure of A. triostegus throughout the tropical Pacific Ocean revealed a strong geographic pattern. Overall, there was significant population differentiation (multilocus F(ST) = 0.199), which was geographically structured according to bootstraps of neighbor-joining analysis on Nei's unbiased genetic distances and AMOVA analysis. The genetic structure revealed five geographic groups in the Pacific Ocean: western Pacific (Guam, Philippines, Palau, and Great Barrier Reef); central Pacific (Solomons, New Caledonia, and Fiji); and three groups made up of the eastern populations, namely Hawaiian Archipelago (north), Marquesas (equatorial), and southern French Polynesia (south) that incorporates Clipperton Island located in the northeastern Pacific. In addition, heterozygosity values were found to be geographically structured with higher values grouped within Polynesian and Clipperton populations, which exhibited lower population size. Finally, the genetic differentiation (F(ST)) was significantly correlated with geographic distance when populations from the Hawaiian and Marquesas archipelagos were separated from all the other locations. These results show that patterns of differentiation vary within the same species according to the spatial scale, with one group probably issued from vicariance, whereas the other followed a pattern of isolation by distance. The geographic pattern for A. triostegus emphasizes the diversity of the evolutionary processes that lead to the present genetic structure with some being more influential in certain areas or according to a particular spatial scale. PMID- 11926507 TI - Species divergence in sexually selected traits: increase in song elaboration is related to decrease in plumage ornamentation in finches. AB - Elaborate plumage and complex songs of male birds are two of the best-known examples of sexually selected traits, yet the interaction between these traits is poorly understood. Theory suggests that among a suite of potential displays, animals will emphasize traits that are most conspicuous, least costly, or best signal condition and reduce display of other traits. Here we examined the relationship between song and plumage elaborations in cardueline finches, songbirds that are highly variable in plumage displays and songs, but that share a similar mating system. We statistically controlled for body mass, habitat characteristics, and phylogenetic relationships and found that across species song complexity was strongly negatively related to elaboration of plumage ornamentation. When plumage coloration was partitioned into carotenoid-based and melanin-based components, song complexity was negatively related to elaboration of male carotenoid-based coloration but unrelated to elaboration of melanin-based coloration. These observations support the idea that, for condition-dependent traits, animal species trade off trait expression in response to changes in the costs or the information content of these traits. We discuss several alternative explanations for the observed pattern. PMID- 11926508 TI - Climbing to reach females: Romeo should be small. AB - The race for reaching mates by the time they are receptive, or sexual selection by scramble competition, has received little attention. We argue that smaller males are favored in species in which the male must climb to reach females located in high habitat patches. This new explanation we term the "gravity hypothesis" of sexual size dimorphism (SSD). We show that a simple biomechanical model of animal movement predicts that: (1) selection should favor a comparatively smaller size in the searching sex when searching involves climbing; and (2) this effect should be stronger in larger species than in smaller species. In reaching high habitats, smaller, faster searchers will be favored either through sexual selection by scramble competition and/or by escaping predation easier by running faster on vertical surfaces. Different spider species are found at a wide range of heights. We compiled a dataset of spider taxa and arranged their habitats according to four height categories, ranked from soil surface to trees. We show that, after controlling for phylogeny, both predictions of the gravity hypothesis of SSD are met. Thus, it appears that the constraint imposed by gravity on climbing males is a selective factor in determining male dwarfism. PMID- 11926509 TI - Occupational health services for small-scale enterprises in Korea. AB - Korea needs national strategies to handle problems of Small Scale Enterprises (SSEs) systematically. Since 1993, the Korean government has begun to provide financial subsidy programs for Occupational Health Services (OHSs) in SSEs from Occupational Injury Prevention Fund. To identify the health care status in SSEs in Korea, 5,080 factories, which had participated in the Government-funded Subsidy Program in 1997, were surveyed. The overall morbidity of the workers in these SSEs was higher than the national average for both general and occupational diseases. Based on the health examinations for occupational disease of those workers exposed to occupational hazards such as noise, dust, or solvents, we could find the industry-specific occupational disease patterns. From this result, we would plan the targeted occupational health services to specific groups. In spite that the effectiveness of this program is not completely assessed, our results indicate that it is desirable for this program to be continued in Korea. In addition, this program may be a good model for rapidly developing countries. PMID- 11926510 TI - Computerized posturography with sway frequency analysis: application in occupational and environmental health. AB - To examine the effects of occupational and environmental neurotoxicants on vestibular, cerebellar and spinocerebellar functions, the following three groups of subjects were examined, using a computerized posturography with sway frequency analysis: (1) 49 male chemical factory workers exposed to lead stearate, aged 27 63 (mean 43) years, with concurrent blood lead concentrations (BPbs) of 11-113 (mean 48) microg/100 g and past mean BPbs of 7-52 (mean 24) microg/100 g; (2) 29 male sandal, shoe and leather factory workers, aged 35-73 (mean 51) years, with urinary 2,5-hexanedione (HD) concentrations of 0.41-3.06 (mean 1.20) mg/g creatinine; and (3) 9 females, aged 19-58 (mean 29) years, who were exposed to sarin accidentally 6-8 months before the study (Tokyo Subway Sarin Poisoning, March 20,1995) and showed plasma cholinesterase (ChE) activities of 13-95 (mean 68) IU/l on the day of poisoning. The pattern of posturographic changes in lead workers suggested that the vestibulocerebellum (lower vermis), anterior cerebellar lobe and spinocerebellar afferent pathway were asymptomatically affected; the vestibulocerebellar change reflected concurrent lead absorption and the anterior cerebellar one reflected past absorption. Similarly, vestibulocerebellar and spinocerebellar functions were affected by n-hexane in solvent workers; the effect on the vestibulocerebellar function was probably inhibited by xylene. Also, the chronic (long-term) effect on the vestibulocerebellar function persisted in acute sarin poisoning. It is thus suggested that the vesitibulocerebellar function is most sensitive to all the three chemicals examined. It appears that the computerized posturography with frequency analysis is a useful technique for assessment of vestibular, cerebellar and spinocerebellar effects in occupational and environmental health. PMID- 11926511 TI - Evaluation of activity of erythrocyte pyrimidine 5'-nucleotidase (P5N) in lead exposed workers: with focus on the effect on hemoglobin. AB - Anemia that accompanies lead poisoning is in part the result of various inhibitory effects of lead on heme biosynthesis. Lead also increases the rate of red blood cell destruction due to the profoundly depressed activities of erythrocyte pyrimidine 5'-nucleotidase (P5N) activities. We studied parameters of the two metabolic pathways in the workers exposed to lead to evaluate P5N in the lead exposed workers and which pathway has an effect on hemoglobin (Hb) level. 29 male workers in the secondary lead smelting as high exposure group, 46 male workers in the manufacturer of inorganic pigment as low exposure group and 56 clerical male workers from another plant as non-exposed group were studied. Activity of P5N, lead concentration in whole blood (PbB), zinc protoporphyrin (ZPP), Hb, and ferritin were determined. In the present study, P5N activity of nucleotide metabolic pathway correlated with Hb after controlling indices of iron deficiency anemia (ferritin) occurring concurrently and heme biosynthetic pathway (ZPP) in the high exposure group while heme biosynthetic pathway did not correlate with Hb after controlling other two variables in exposure groups. These findings suggest that P5N rather than heme biosynthetic pathway has a major effect on Hb level even in workers without manifest hemolytic anemia. PMID- 11926512 TI - Distribution and cardiovascular risk correlates of serum triglycerides in young Japanese adults. AB - To examine the distribution and cardiovascular risk correlates of serum triglycerides, a cross-sectional population study based on annual health examinations at the workplace was performed in 2199 young Japanese adults aged 23 to 37 years. Triglyceride levels showed significant sex (male > female) differences, and the percentages of those with high triglycerides (> or = 150 mg/dl) were 9.4% for males and 0.8% for females. In terms of conjoint trait of dyslipidemia, 86.1% of males displayed normal levels of both triglycerides (< 150 mg/dl) and high-density lipoprotein (HDL) cholesterol (> or = 40 mg/dl), while 98.7% of females had normal values. Age- and sex-specific triglyceride levels above the 75th percentile (equivalent to 82-116 mg/dl for males and 56-63 mg/dl for females) increased the risk (odds ratio (OR)) for having obesity, hypertension, and hyperuricemia by 2.9 (95% confidence interval (CI)=2.0-4.3),1.7 (CI=1.1-2.9), and 3.0 (CI=1.6-5.9), respectively. The respective ORs for triglyceride levels above the 75th percentile and HDL cholesterol below the 25th percentile (equivalent to 45-49 mg/dl for males and 58-63 mg/dl for females), compared with triglyceride levels the 75th percentile or less and HDL cholesterol levels the 25th percentile or more, were 8.7 (CI=5.8-12.9), 2.2 (CI=1.5-3.3), and 6.0 (CI=3.2-11.5). Our results suggest a threshold effect of triglyceride levels considered as normal on enhanced cardiovascular risk in young Japanese adults, especially in those with low HDL cholesterol levels. PMID- 11926513 TI - A cross-sectional survey of 32 workers exposed to hexahydrophthalic and methylhexahydrophthalic anhydrides. AB - The relation between exposure and sensitization or the appearance of symptoms of the eyes and airways was investigated in a cross-sectional study on 32 workers from a plant using epoxy resin with a mixture of hexahydrophthalic anhydride (HHPA) and methylhexahydrophthalic anhydride (MHHPA) as a hardener. The main component in the hardener was HHPA, and the geometric mean concentrations of HHPA in the workplaces were extremely low (<40 microg/m3) in recent years, compared to the Occupational Exposure Limit-Ceiling for phthalic anhydride (2 mg/m3). However, specific IgE antibody to HHPA was detected in serum from 8 (25%) out of the workers: of those, 5 workers experienced symptoms of the eyes and nose during work (group sensitized symptomatic (SS)) and 3 workers did not (group sensitized nonsymptomatic (SN)). The other 24 workers had no signs of sensitization and did not complain of work-related symptoms. Based on occupational history and anamnestic data, it was concluded that one subject in the SS group and all the subjects in the SN group had been sensitized by higher exposures in the past. The symptoms of 4 subjects in the SS group occurred only when carrying out short time, particular tasks (15-30 min) a few times a day, such as the resin mixing procedures, manual application of the resin, or opening of ovens. High peak exposures were estimated to have occurred during the particular tasks. Our results suggest that short-time peak exposures may have a great impact on the development of specific IgE or work-related symptoms. Therefore, to minimize the risk of sensitization and work-related symptoms, a reduction of exposure during particular tasks with high peak exposures, along with a decrease in mean 8-h time weighted average exposure, should be achieved. PMID- 11926514 TI - Skin problems among fiber-glass reinforced plastics factory workers in Japan. AB - Two surveys, one in winter the other in summer time, examined the skin problems of the entire manual workers (N=148) from 11 small-to-medium sized fiber-glass reinforced plastics (FRP) factories located in Kyushu, Japan. The workers were exposed to unsaturated polyester resin, including styrene and auxiliary agents such as cobalt naphthenate, hardeners such as methyl ethyl ketone peroxides, glass fiber and dust including shortened glass fiber and plastic particles. Eighty-seven workers (58.8%) reported having skin problems (mainly itching or dermatitis) since they started to work in FRP manufacturing and 25 workers had consulted a physician because of their skin problems; one worker was forced to take sick leave because of his severe dermatitis. History of allergic diseases and shorter occupational period (duration of employment) in a FRP factory were associated with greater probability of having a history of work-related skin symptoms. Workers in factories where dust-generating and lamination sites were located in different buildings were significantly less likely to have a history of skin problems than those in factories where the two sites were located in the same building. Of the 67 workers examined in both seasons closed to double the prevalence of dermatitis was found in summer (23.3%) than winter (13.4%). PMID- 11926515 TI - Successful experiences with a handy equipment to measure dust concentrations in various workplaces. AB - Performance of a direct-reading handy equipment for dust counting in workroom air, PDR (MIE, USA), was field-tested in comparison with the results of low volume air sampling-gravimetry. Application in 64 workplaces of various type dust work with dust concentrations of up to 1.6 mg/m3 showed that the agreement was generally good with a correlation coefficient (r) >0.64. The calculated regression line had a slope close to one, and the intercept on the axis was next to nil, suggesting that the measurement with PDR is essentially the same with that by low-volume air sampling. Further analysis by types of dust work disclosed that the correlation was best when examined in 12 foundries with r as large as 0.95. In contrast, r was small (0.52) in 11 welding workplaces, possibly due to smaller particle size of dust generated during welding. PMID- 11926516 TI - Prevalence of skin disease among nursing home staff in southern Taiwan. AB - To establish the prevalence of skin disease among nursing home workers in southern Taiwan, dermatological examination was performed on 75 nursing home staff from 11 institutions in Tainan county. Fungal infections were the most common skin diseases identified, affecting 21.4% of all employees. Other conditions included xerosis (13.3%), scabies (10.7%) and dermatitis (8.0%). Fungus was found mainly on the feet and hands (68.7% and 31.3% of all fungal cases respectively). Most xerosis sites were identified on the lower leg (90.0% of all xerosis cases), while all workers with scabies had the disease on their forearm. Dermatitis was diagnosed predominately on the forearm (50.0% of all dermatitis cases). The prevalence of fungus and scabies was higher than other studies, while dermatitis occurred less frequently than previous reports. Although not statistically significant, we believe that wet work and occupational contact with nursing home patients may have been important risk factors for these conditions. PMID- 11926517 TI - A study of the hand-arm vibration syndrome in Okinawa, a subtropical area of Japan. AB - The purpose of this study was to understand the peculiarity of the development of Hand-Arm Vibration Syndrome (HAVS) in a subtropical area of Japan. We analyzed the medical records of 21 subjects reported by the Okinawa Labor Bureau, and tried to determine whether the warm environment has any effect on the severity of HAVS. The mean operating time of vibration tools for all the subjects was over 10,000 hrs. Almost all the subjects from Okinawa had been working only in Okinawa. The presence of Vibration Induced White Fingers (VWF) was markedly smaller in the subjects from Okinawa as compared to those from other prefectures. The results of Cold Water Immersion Tests were similar for all subjects. Ten minutes after the Cold Water Immersion Test, the mean finger skin temperature was about 19 degrees C in all subjects. These findings suggest that not only the operating time of vibration tools but also the warm environment might have an effect on the severity of HAVS. However, the number of subjects in our study was very small. There is the need therefore for further investigations with a larger number of subjects. PMID- 11926518 TI - Occupational dermatitis from a one-component naphthalene type epoxy adhesive. AB - In an electronics plant, a new one-component naphthalene type epoxy resin was used as an adhesive for reinforcing a circuit board. The resinous part of the adhesive consisted of diglycidyl ether of bisphenol A (DGEBA) and 1,6-bis(2,3 epoxypropoxy)naphthalene type epoxy resins. The hardener was methylhexahydrophthalic anhydride (MHHPA). Of 54 workers, 15 (27.8%) were diagnosed to have work-related dermatitis but were not patch tested. Therefore, it was impossible to determine the specific agent responsible for the worker's symptoms or to distinguish between allergic and irritant contact dermatitis. They worked without protective gloves until they started to develop skin symptoms. The hands were the commonly affected region (13 out of 15 cases). The latent period of dermatitis was very short (mean 2.2 weeks). Of these, 10 cases (66.7%) received medication for dermatitis, and 9 cases (60%) were transferred to other work. The work-related skin symptoms were closely related to the specific tasks, i.e., filling dispensers with the adhesive and manual application of the adhesive to a portion of a circuit board using a dispenser. For occupational hygiene reasons, contact with epoxy resins should be minimized by taking all possible measures into use, including protective gloves. Further studies are required to clarify the allergenicity of 1,6-bis(2,3-epoxypropoxy)naphthalene, since very little is known about the mechanism through which it leads to the symptoms of dermatitis. PMID- 11926519 TI - Lifetime principal occupation and risk of cognitive impairment among the elderly. AB - We used a nested case-control design with study participants sampled from two cohorts, for a total of 2,198 elderly people 65 years or older and completed cognitive tests between 1993 and 1997, to assess the association between an individual's lifetime principal occupation and the subsequent risk of cognitive impairment. Cases consisted of 290 older adults with impaired cognitive functioning. For each case, two controls with comparable age (within 5 years) and sex frequencies were randomly sampled from the seniors free of cognitive impairment. Occupational data were collected through interviews. Individual's job content was coded into one of the occupational categories or the occupation-based social classes. Compared to those who were former legislators, government administrators, or business executives and managers, a significantly elevated risk of cognitive impairment was estimated for those who were employed as agriculture/animal husbandry/forestry/fishing workers (odds ratio (OR)=3.2), craft and related trades workers (OR=2.2), plant and machine operators and assemblers (OR=14.7), workers of elementary occupations (OR=3.2), or housekeepers (OR=2.6). We also observed health inequalities in the risk of cognitive impairment across social classes with a significant dose-response trend in which unskilled blue-collar workers had the highest risk. After adjustment for education, we still observed an inverse relationship between risk of cognitive impairment and occupational class. This may mean that lifetime longest-held occupation is more intimately involved in the causal pathways leading to cognitive impairment. Further studies that collect information on specific work hazards would help make specific interpretations of the observed effect of lifetime longest-held occupation in early adulthood on risk of cognitive decline in late life. PMID- 11926520 TI - Are hydrophobins and/or non-specific lipid transfer proteins responsible for gushing in beer? New hypotheses on the chemical nature of gushing inducing factors. AB - Gushing of beer is characterised by the fact that immediately after opening a bottle a great number of fine bubbles are created throughout the volume of beer and ascend quickly under foam formation, which flows out of the bottle. This infuriating gushing phenomenon has been, and still is, a problem of world-wide importance to the brewing industry. It is generally assumed that the causes of malt-derived gushing are due to the use of "weathered" barley or wheat and the growth of moulds in the field, during storage and malting. We now develop a hypothesis connecting several lines of evidence from different laboratories. These results indicate that the fungal hydrophobins, hydrophobic components of conidiospores or aerial mycelia, are gushing-inducing factors. Furthermore, increased formation of ns-LTPs (non-specific lipid transfer proteins), synthesised in grains as response to fungal infection, and their modification during the brewing process may be responsible for malt-derived gushing. PMID- 11926522 TI - Differences in the activity of superoxide dismutase, polyphenol oxidase and Cu-Zn content in the fruits of Gordal and Manzanilla olive varieties. AB - Activity of the enzymes superoxide dismutase (SOD) and polyphenol oxidase (PPO) as well as Cu-Zn content have been monitored during the thirteen weeks growth of both Gordal and Manzanilla olive variety fruits. These metalloenzymes, with Cu and Zn in the prostetic group, are involved in controlling the redox balance in the chloroplast environment. The results indicated that, under similar phenological and environmental conditions, there are periodic peaks of SOD activity in both varieties, followed by fluctuations in the copper content of the fruit. This was interpreted as a common and simultaneous response to situations of oxidative stress, and this response was more intense in the variety Gordal. The enzyme PPO showed an activity peak at start of growth and then practically disappeared. Thus, its activity cannot be correlated with situations of stress or with changes of Cu and Zn in the fruit. PMID- 11926521 TI - Pharmaco-toxicological study of Kageneckia oblonga, Rosaceae. AB - The probable antipyretic, antiinflammatory, analgesic and antioxidant properties of Kageneckia oblonga, Rosaceae, were investigated and the major compounds of its active extracts were isolated. The study comprised the acute toxicity of the extracts of global methanol, hexane, dichloromethane and methanol. The cytotoxicity of global methanol extract was studied in three tumoral cell lines. All the extracts exhibited the pharmacological activities under study. Methanol and dichloromethane were the most toxic extracts. From the global methanol extract, isolations were performed of prunasin, 23,24- dihydro-cucurbitacin F, and a new cucurbitacin, 3beta-(beta-D-glucosyloxy)-16alpha,23alpha-epoxycucurbita 5,24-diene-11-one. The cytotoxicity of both cucurbitacins on human neutrophils at the assayed concentrations was not statistically significant. In-vitro assays showed that both cucurbitacins can be partly responsible for the analgesic, antipyretic, and anti-inflammatory activities. Evaluation was done of the cytotoxicity of global methanol extract, 23, 24-dihydrocucurbitacin F, aqueous extracts and prunasin against P-388 murine leukaemia, A-549 human lung carcinoma and HT-29 colon carcinoma. Since global methanol extract presented a strong cytotoxicity against P-388 murine leukaemia, A-549 human lung carcinoma, and HT 29 cell lines, it is highly probable that this extract contain one or more cytotoxic compounds that could be investigated for their potential use as an agent against cancer. PMID- 11926524 TI - Comparative X-ray studies on the interaction of carotenoids with a model phosphatidylcholine membrane. AB - The interaction of structurally different carotenoids with a membrane molecular model was examined by X-ray diffraction. The selected compounds were beta carotene, lycopene, lutein, violaxanthin, zeaxanthin, and additionally carotane, a fully saturated derivative of beta-carotene. They present similarities and differences in their rigidity, the presence of terminal ionone rings and hydroxy and epoxy groups bound to the rings. The membrane models were multibilayers of dipalmitoylphosphatidylcholine (DPPC), chosen for this investigation because the 3 nm thickness of the hydrophobic core of its bilayer coincides with the thickness of the hydrophobic core of thylakoid membranes and the length of the carotenoid molecules. Results indicate that the six compounds induced different types and degrees of structural perturbations to DPPC bilayers in aqueous media. They were interpreted in terms of the molecular characteristics of DPPC and the carotenoids. Lycopene and violaxanthin induced the highest structural damage to the acyl chain and polar headgroup regions of DPPC bilayers, respectively. PMID- 11926523 TI - Induction of anthranilate synthase activity by elicitors in oats. AB - Oat phytoalexins, avenanthramides, are a series of substituted hydroxycinnamic acid amides with anthranilate. The anthranilate in avenanthramides is biosynthesized by anthranilate synthase (AS, EC 4.1.3.27). Induction of anthranilate synthase activity was investigated in oat leaves treated with oligo N-acetylchitooligosaccharide elicitors. AS activity increased transiently, peaking 6 h after the elicitation. The induction of activity was dependent on the concentration and the degree of polymerization of the oligo-N acetylchitooligosaccharide elicitor. These findings indicate that the induction is part of a concerted biochemical change required for avenanthramide production. The elicitor-inducible AS activity was strongly inhibited by L-tryptophan and its analogues including 5-methyl-DL-tryptophan, and 5- and 6-fluoro-DL-tryptophan, while the activity was not affected by D-tryptophan. The accumulation of avenanthramide A was also inhibited by treatment of elicited leaves with these AS inhibitors, indicating that a feedback-sensitive AS is responsible for the avenanthramide production. In elicited leaves, the content of free anthranilate remained at a steady, low level during avenanthramide production. Moreover, administration of anthranilate to elicited oat leaves resulted in an enhanced avenanthramide accumulation. AS may play a role as a rate-limiting enzyme in the biosynthesis of avenanthramides. PMID- 11926525 TI - Influence of different light intensities on the content of diosgenin, lipids, carotenoids and fatty acids in leaves of Dioscorea zingiberensis. AB - Cultivation of the climbing plant Dioscorea zingiberensis at a light intensity of 100 microE. m(-2) sec(-1) yields three different phenotypes. Most of the plants grow as green phenotype (DzW). Two further forms differ in their leaf shape and leaf color. Whereas one type exhibits a more pointed leaf shape in the upper part of the plant with leaves appearing yellow-green with white stripes or hatchings (DzY), the other type shows a more round leaf shape with an intensive yellow green color (DzT). These three plant types differ in their diosgenin content not only in their rhizomes but also in the chloroplasts. In the rhizomes the diosgenin content in the green form is 0.4%, in the DzY-form 0.6% and in the DzT form even 1.3% of the dry weight. Furthermore, even in chloroplasts of the green DzW-form and of the DzY-form the presence of diosgenin was demonstrated. It occurs there as the epimeric form yamogenin. The DzT-form contains no yamogenin in its chloroplasts. Besides this, these plant forms differ in their chlorophyll and carotenoid content and in their fatty acid composition. Carotenoids increase from 1.3% of total lipids in the green phenotype to 3.3% in the DzY- and to 4.2% in the DzT-form. This increase refers to beta-carotene as well as to lutein and neoxanthin. The chlorophyll content in the green type is 8.1% and lower in the DzY-form with 7%. The highest chlorophyll content is found in the DzT-form with 12%. Fatty acids in the DzY-form and in the DzT-form have a more unsaturated character than in the green phenotype. The content of the monoenoic acid trans hexadecenoic acid is considerably lower in both phenotypes when compared to the green phenotype. In both phenotypes the quantity of fatty acids with 16 carbon atoms is reduced, whereas fatty acids with 18 carbon atoms occur in higher concentration. Cultivation of the green phenotype (DzW) at the three light intensities of 10, 100 and 270 microE x m(-2) x sec(-1) leads to changes of the diosgenin content in rhizomes, to an increase of leaf dry weight, to a reduction of the grana structure in chloroplasts and therewith to a decrease of the chlorophyll content. The total lipid content is highest under the cultivation at 100 microE x m(-2) x sec(-1) and reduced by 30% at 10 and 270 microE x m(-2) x sec(-1). Carotenoids, however, are highest in shaded plants (10 microE x m(-2) x sec(-1)) and plants grown under high light conditions of 270 microE x m(-2) x sec(-1). At 100 microE x m(-2) x sec(-1) a decrease of saturated fatty acids is observed in comparison to plants grown under shaded conditions. PMID- 11926526 TI - Phytoecdysteroid levels and distribution during development in Limnanthes alba Hartw. ex Benth. (Limnanthaceae). AB - Phytoecdysteroid (PE) production and accumulation in Limnanthes alba Hartw. ex Benth. is associated with flowering. PE content per plant remains fairly constant during the primary growth phase of the plant and only begins to increase significantly above amounts found in the seed once the development of the flower stalk has begun. Both content and concentration increase concomitantly from this point. Distributions in individual plants also associated the highest levels of PE accumulation with the reproductive tissues. This substantiates the association of PE with tissues of greatest fitness value and therefore the hypothesis that they contribute to defence. Analysis of extracts of L. alba tissues by reversed phase HPLC coupled with ecdysteroid-specific RIA was used to monitor ecdysteroid profiles. RIA-positive peaks co-chromatographing with 20-hydroxyecdysone, ecdysone and ponasterone A were detected and several tissues also contain PE conjugates. Seedmeal of L. alba appears to be a convenient and promising source for the commercial isolation of the potent PE ponasterone A. PMID- 11926527 TI - Monobromoisophakellin, a new bromopyrrole alkaloid from the Caribbean sponge Agelas sp. AB - A detailed analysis of the chemical constituents of a Caribbean specimen of Agelas sp. was carried out. Four brominated compounds (1-4) were isolated and one of them was identified as a new bromopyrrole metabolite, monobromoisophakellin (1). The structure of 1 was determined using spectroscopic methods. All compounds were tested for their antifeedant activity against the Caribbean reef fish Thalassoma bifasciatum in an aquarium assay. PMID- 11926528 TI - Bromosceptrin, an alkaloid from the marine sponge Agelas conifera. AB - Six dimeric bromopyrrole alkaloids (1-6) were isolated from a Florida Keys specimen of Agelas conifera. One of the constituents was identified as a new bromopyrrole metabolite, bromosceptrin (1). The structure of 1 was established from MS spectrometry and 1D and 2D NMR spectrocopy. PMID- 11926529 TI - Chemical defense and antifouling activity of three Mediterranean sponges of the genus Ircinia. AB - The defense roles and the antifouling activity of the organic extracts and the major metabolites of the sponges Ircinia oros, I. variabilis and I. spinosula were investigated. The antifeedant activity was tested in experimental aquaria on the generalist predator fish Thalassoma pavo as well as in coastal ecosystems rich in fishes. Some of the major metabolites exhibited high levels of antifeedant activity. The antifouling activity was tested in laboratory assays, against representatives of the major groups of fouling organisms (marine bacteria, marine fungi, diatoms, macroalgae and mussels). All extracts showed promising levels of activity. As was expected, no single extract was active in all tests and some fractions that were effective against one organism showed little or no activity against the others. The high but variable level of antifouling activity in combination with the absence of toxicity (tested on the development of oyster and sea urchin larvae) shows the potential of these metabolites to become ingredients in environmentally friendly antifouling preparations. PMID- 11926530 TI - Barbeyol: a new phenolic indane type component from Barbeya oleoides. AB - The aerial parts of Barbeya oleoides Schweinf (Family: Barbeyaceae) has afforded a new phenolic indane type component, which has been characterized as (6R, 4aR, 5aR)-5, 4a, 5a-trihydroindeno- (1, 2-a) indane-2, 4, 9-triol-6-O-beta-acetate (1) on the basis of spectral analysis and has been designated as barbeyol. PMID- 11926531 TI - Glutathione peroxidase from the liver of Japanese sea bass Lateolabrax japonicus. AB - Glutathione peroxidase (EC 1.11.1.9) present in the liver of Japanese sea bass (Lateolabrax japonicus) was extracted and purified by phenyl-toyopearl 650M, butyl-toyopearl 650M and DEAE-toyopearl 650M column chromatography. The molecular weight of the enzyme was estimated to be about 70 kDa by gel filtration by toyopearl HW-55F. On SDS-PAGE, this enzyme was composed of two identical subunits with 35 kDa and was a dimer. This enzyme was a typical SH-enzyme that was inhibited by iodoacetic acid, PCMB, DTNB, and Hg. PMID- 11926532 TI - 1,4-benzoquinone attracts males of Rhizotrogus vernus Germ. AB - Two candidate attractants, phenol and 1,4-benzoquinone, a synthetic mixture of typical compounds from green-leaf odours [(Z)-3-hexenyl acetate: (Z)-3-hexen-1 ol: benzaldehyde: (E)-2-hexen-1-ol: 1-hexanol; 100:20:10:1:1] and freshly damaged oak leaves were screened for field attractancy in funnel traps in Hungary. Males of two Rhizotrogus spp. (Coleoptera, Scarabaeidae, Melolonthinae), R. aestivus 01. and R. vernus Germ. were caught in larger numbers. While R. aestivus catches were probably chance captures, male R. vernus was significantly attracted to the baits containing 1,4-benzoquinone. This compound represents a promising basis for the development of a monitoring trap for R. vernus. PMID- 11926533 TI - Effects of L-phenylalanine on acetylcholinesterase and Na+,K+-ATPase activities in suckling rat frontal cortex, hippocampus and hypothalamus. AB - The effect of different L-phenylalanine (Phe) concentrations (0.12-12.1 mM) on acetylcholinesterase (AChE), (Na+,K+)-ATPase and Mg2+-ATPase activities was evaluated in homogenates of suckling rat frontal cortex, hippocampus and hypothalamus. Phe, at high concentrations, reduced AChE activity in frontal cortex and hippocampus by 18%-20%. On the contrary, the enzyme activity was unaltered in the hypothalamus. Na+,K+-ATPase was stimulated by high levels of the amino acid, both in the frontal cortex and the hypothalamus by 60%, whereas it was inhibited in the hippocampus by 40%. Mg2+-ATPase was not influenced by Phe. It is suggested that: a) In the frontal cortex, the improper acetylcholine (ACh) release, due to AChE inhibition by Phe, combined with the stimulation of Na+,K+ ATPase, possibly explain tremor and the hyperkinetic behaviour in patients with classical phenylketonuria (PKU). b) In the hippocampus, inhibition of AChE by Phe could lead to problems in memory, while Na+,K+-ATPase inhibition by Phe may induce metabolic disorders and electrical instability of the synaptosomal membrane. c) In the hypothalamus, the behavioral problems in PKU "off diet" may be related to noradrenaline (NA) levels, which are probably correlated with the modulated Na+,K+-ATPase by Phe. PMID- 11926534 TI - Phylogenetic relationship of Bombyx mori protein disulfide isomerase. AB - A cDNA that encodes protein disulfide isomerase was isolated from Bombyx mori (bPDI), in which an open reading frame of 494 amino acids contained two PDI typical thioredoxin active sites of WCGHCK and an ER retention signal of the KDEL motif at its C-terminal. The bPDI protein shared less than 55% of the amino acid sequence homology with other reported PDIs. bPDI is most genetically similar to the D. melanogaster PDI. The most serious evolutional diversity was observed between the metazoa and nematoda through PDI evolutional processing. Although bPDI shows a relatively low amino acid homology with other PDIs, in which both sites of the two thioredoxin active sites and the endoplasmic reticulum (ER) retention signal are completely conserved, it was successfully recognized by anti rat PDI antibodies. This suggests that bPDI may have the activity of a protein isomerase and a chaperone. PMID- 11926535 TI - Effect of Cu/Zn-superoxide dismutase from the fungal strain Humicola lutea 103 on antioxidant defense of Graffi tumor-bearing hamsters. AB - A novel Cu/Zn-containing superoxide dismutase (SOD) was isolated from the fungal strain Humicola lutea 103. Previously, a protective effect of this enzyme (HLSOD) against tumor growth and also superoxide production in Graffi tumor-bearing hamsters (TBH) were established. The aim of the present study was to investigate the effect of HLSOD on the activity of endogenous SOD and catalase in the cells from TBH during tumor progression. Our results point out that transplantation of Graffi tumor causes a significant decrease in SOD activity in the cells from liver of the hosts (from 35 to 59% compared to the control). In the tumor cells relatively low levels of SOD (about 7 U mg protein(-1)) were found, and Cu/ZnSOD was the main isoenzyme in total SOD activity. Tumor growth resulted in a reduction of catalase activity, which correlated with the process of tumor progression. A single dose (65 U) treatment with HLSOD caused an increase in endogenous SOD and catalase activity in healthy animals and resulted in restoration of the antioxidant ability in liver cells of the hosts at the early stage of tumor progression. The results show the possible participation of HLSOD in the host oxidant-antioxidant balance, which is probably one of the factors of its immunoprotective action established earlier. PMID- 11926537 TI - Cucurbitacin F in seeds of Kageneckia angustifolia (Rosaceae). AB - Cucurbitacin F and the cyanogenetic compounds prunasin were isolated and identified from the seeds of Kageneckia angustifolia. PMID- 11926536 TI - (E,E)-alpha-farnesene the main substance of the volatiles of the flowers from European mistletoe (Viscum album L.). AB - (E,E)-alpha-Farnesene was extracted as the main component of the volatile fraction of male Viscum album L. Male and female flowers of V. album L. growing on different host trees were analysed by solid phase microextraction and by lipophilic extraction. PMID- 11926538 TI - New triterpenoidal alkaloids from Buxus sempervirens. AB - Phytochemical studies on the ethanolic extract of the roots of Buxus sempervirens of Turkish origin have resulted in the isolation of two new triterpenoidal alkaloids, (+)-16a, 31-diacetylbuxadine (1), (-)-Nb-demethylcyclomikuranine (2) along with three known natural products, (-)-cyclomikuranine (3), (-) cyclobuxophylline-K (4) and (+)-buxaquamarine (5) isolated for the first time from this species of genus Buxus. The structures of these new natural products were established on the basis of extensive spectroscopic studies. Compound 1 exhibited antibacterial activity against human pathogenic bacteria and weak phytotoxic activity against Lemna minor Linn. PMID- 11926539 TI - Lignans from leaves of Rollinia mucosa. AB - A new furofuranic lignan named (+)-epimembrine together with known (+) epieudesmine and (+)-epimagnoline were isolated from leaves of R. mucosa. Their structures were determined by spectroscopic data. Palmitone and a mixture of beta sitosterol and stigmasterol were also isolated. PMID- 11926540 TI - Two new sulfated furostanol saponins from Tribulus terrestris. AB - The known furostanol saponins methylprotodioscin and protodioscin and two new sulfated saponins, sodium salt of 26-O-beta-glucopyranosyl-22alpha-methoxy-(25R) furost-5-ene-3beta,26-diol-3-O-alpha-rhamnopyranosyl-(1-->2)-beta-4-O-sulfo glucopyranoside (methylprototribestin) and sodium salt of 26-O-beta glucopyranosyl-22alpha-hydroxy-(25R)-furost-5-ene-3beta,26-diol-3-O-alpha rhamnopyranosyl-(1-->2)-beta-4-O-sulfo-glucopyranoside (prototribestin) have been isolated from the aerial parts of Tribulus terrestris L. growing in Bulgaria. The structures of the new compounds were elucidated on the basis of 1D and 2D (DQF COSY, TOCSY, HSQC-TOCSY, HSQC, HMBC, ROESY) NMR data, ESI mass spectra and chemical transformation. PMID- 11926541 TI - Prenylated p-coumarates from the twigs of Phebalium rude subsp. amblycarpum (Rutaceae). AB - Phebarudol, a novel prenylated p-coumarate, was isolated from the twigs of Phebalium rude Bartl. subsp. amblycarpum (F. Muell.) P. G. Wilson (Rutaceae) together with the two already known related compounds, werneria chromene and methyl demethoxywutaiensate. The structure of phebarudol was established by spectroscopic methods. PMID- 11926542 TI - Iridoid patterns of genus Plantago L. and their systematic significance. AB - The distribution of 14 iridoid glucosides in 14 Plantago L. species (44 samples corresponding to 18 taxa) was shown. P. tenuiflora and P. gentianoides were studied for iridoids for the first time. The iridoid patterns showed a good correlation with morphological and other chemical features of the representatives of genus Plantago. The studied species are grouped together according to the iridoid patterns: species containing mainly aucubin (P. major, P. cornuti, P. gentianoides); species containing aucubin and aucubin derivatives (P. subulata, P. media); species containing aucubin and catalpol (P. lanceolata, P. altissima, P. argentea, P. lagopus, P. atrata); species containing aucubin and plantarenaloside (P. afra, P. scabra). PMID- 11926544 TI - Alkylresorcinols in barley (Hordeum vulgare L. distichon) grains. AB - This study was carried out to compare grains of barley (Hordeum vulgare L. distichon) regarding contents and compositions of 5-n-alkylresorcinols. Mixtures of resorcinol homologues were isolated from acetone extracts from five barley cultivars. These polyketide metabolites were identified by chromatographic and spectroscopic means. The content and homologue patterns among different varieties were similar. The predominant compounds were 1,3-dihydroxy-5-n-heneicosylbenzene (C21:0), 1,3-dihydroxy-5-n-nonadecylbenzene (C19:0) and 1,3-dihydroxy-5-n pentacosylbenzene (C25:0). The alkylresorcinol concentrations, in contrast to their compositions, depended on environmental and agricultural factors. PMID- 11926543 TI - Lipid analysis of Greek walnut oil (Juglans regia L.). AB - The walnut oil (Juglans regia L.) total lipids (TL) were extracted by the Bligh Dyer method and the lipid classes have been isolated by chromatographic techniques and they were analyzed by high performance thin layer chromatography (HPTLC)/FID and GC-MS. The oil was found to be rich in neutral lipids (96.9% of total lipids) and low in polar lipids (3.1% of total lipids). The neutral lipid fraction consisted mainly of triacylglycerides whereas the polar lipids mainly consisted of sphingolipids. GC-MS data showed that the main fatty acid was linoleic acid. Unsaturated fatty acids were found as high as 85%, while the percentage of the saturated fatty acids was found 15%. Two types of liposomes were prepared from the isolated walnut oil phospholipids and characterized as new formulations. These formulations may have future applications for encapsulation and delivery of drugs and cosmetic active ingredients. PMID- 11926545 TI - GLC and GLC-mS analysis of thiophene derivatives in plants and in in vitro cultures of Tagetes patula L. (Asteraceae). AB - The occurrence of thiophenic compounds in diverse plant organs and in in vitro root-, callus- and cell suspension cultures of Tagetes patula cv. Carmen was investigated using capillary GLC and GLC-MS. The separation of thiophenes by capillary GLC and the group specific MS fragmentation with the typical sulfur isotope peaks allowed the unequivocal assignment of individual thiophenes in complex mixtures, even when occurring in traces and in the presence of different geometrical isomers. The extracts of Tagetes patula cv. Carmen contained the following 8 thiophene compounds: 5-(3-buten-1-ynyl)-2,2'-bithienyl (BBT), 5' methyl-5-(3-buten-1-ynyl)-2,2'-bithienyl (MeBBT), 5-(1-pentynyl)-2,2'-bithienyl (PBT), 5-(4-hydroxy-1-butynyl)-2,2'-bithienyl (BBTOH), 2,2',5,2"-terthienyl (alpha-T), 5-(4-acetoxy-1-butynyl)-2,2'-bithienyl (BBTOAc), 5-methylaceto-5'-(3 buten-1-ynyl)-2,2'-bithienyl (AcOCH2BBT), and 5-(3,4-diacetoxy-1-butynyl)-2,2' bithienyl (BBT(OAc)2). The most complex thiophene profile, including the less common PBT was detected in aerial parts of freshly harvested plant material. Under in vitro conditions only the root cultures, but not callus or cell suspension cultures produced substantial amounts of irregular thiophenes confirming that roots are the main site of thiophene biosynthesis. PMID- 11926547 TI - Secondary metabolites from Centaurea deusta with antimicrobial activity. AB - The aerial parts of Centaurea deusta Ten. afforded in addition to several known compounds, mainly sesquiterpene lactones, one new eudesmanolide and one new elemane derivative. Structures of the new compounds were elucidated by spectroscopic methods. The in vitro antifungal and antibacterial activities of the isolated compounds was tested, using the microdilution method. All compounds tested showed high antifungal activity. PMID- 11926546 TI - Inhibition of very-long-chain fatty acid formation by indanofan, 2-[2-(3 chlorophenyl)oxiran-2-ylmethyl]-2-ethylindan-1,3-dione, and its relatives. AB - Indanofan and its analogs inhibited the elongation of stearoyl- or arachidoyl-CoA by [2-14C]-malonyl-CoA in leek microsomes from Allium porrum. Although the precise mode of interaction of indanofan at the molecular level is not completely clarified by the present study, it is concluded that indanofan and analogs act as inhibitor of the elongase enzyme involved in de novo biosynthesis of fatty acids with an alkyl chain longer than C18, called very-long-chain fatty acids (VLCFAs). For a strong inhibition of VLCFA formation chloro substituents at the benzene ring and the oxirane group were necessary. Furthermore, the greenhouse test showed strong activity for indanofan and its analogs, and the scores coincided with cell-free elongation inhibition. The cell-free assay, however, failed to indicate any activity for an analog having a methylene instead of the oxirane group, while both Digitaria ciliaris and Echinochloa oryzicola were killed with 1 kg a.i./ha. This finding cannot be discussed because the applied use rate of 1 kg a.i./ha is too high to allow for a score differentiation. For high concentrations of this compound additional unknown inhibitory effects may be involved besides fatty acid elongation. PMID- 11926548 TI - In vitro cytotoxicity of some natural and semi-synthetic isocoumarins from Paepalanthus bromelioides. AB - Numerous natural compounds have a potential for therapeutic applications, but may have to be chemically modified to alter toxic side effects. We investigated structural parameters that could affect the cytotoxicity of isocoumarins similar to 9,10-dihydroxy-5,7-dimethoxy-1H-naphtho(2,3c)pyran-1-one (paepalantine 1). Paepalantine 1 has antimicrobial activity, as well as significant in vitro cytotoxic effects in the McCoy cell line. Two other natural and two semi synthetic isocoumarins with similar structures obtained from the capitula of Paepalanthus bromelioides were tested on the same cell line by the neutral red assay. Substitution of the 9 and/or 10-OH group made these compounds less cytotoxic. PMID- 11926549 TI - A chemometric interpopulation study of the essential oils of Cistus creticus L. growing in Crete (Greece). AB - The chemical composition of the essential oils of twenty-five populations of Cistus creticus subsp. creticus L. from the island of Crete (Greece) and their interpopulation variability were analysed in detail by GC-MS. 142 compounds were identified representing an average of 56.8-89.8% of the oil composition. The components are represented here by homologous series of monoterpenes, oxygenated monoterpenes, sesquiterpenes, oxygenated sesquiterpenes, diterpenes, labdane diterpenes, aldehydes, alkanes, esters, fatty acids, ketones, and others. Labdane diterpenes were detected and identified in the essential oils and have been found in high percentage composition. The results from the chemical analysis of the essential oils were submitted to chemometric cluster analysis in order to detect some pattern distribution and to identify which constituents can differentiate the groups of individuals. Two main chemotypes (clusters) were well differentiated; the first deals with eight populations of West Crete and the second with the rest of the populations. Cluster analysis based on labdane type diterpenes patterns, proved to be the best chemotype for the examined populations among the other chemical groups. PMID- 11926550 TI - The structures of the pyoverdins from two Pseudomonas fluorescens strains accepted mutually by their respective producers. AB - From Pseudomonas fluorescens PL7 and PL8 structurally related pyoverdins were isolated and their primary structures were elucidated by spectroscopic methods and degradation reactions. Despite of some structural differences both Fe(III) complexes are taken up by either strain with a high rate. The implications regarding the recognition at the cell surface are discussed. PMID- 11926551 TI - Cantleyoside-dimethyl-acetal and other iridoid glucosides from Pterocephalus perennis--antimicrobial activities. AB - Cantleyoside-dimethyl-acetal (6), was isolated from the endemic Greek plant Pterocephalus perennis subsp. perennis in addition to five other known iridoid glucosides, loganin, loganic acid, cantleyoside, secologanin, and secologanin dimethyl-acetal. The structure of these compounds was determined by all spectroscopic means mainly by NMR and MS techniques. The above compounds as well as their acetyl derivatives were tested against six Gram positive and negative bacteria and three pathogenic fungi. PMID- 11926552 TI - Long-term follow-up of achalasia patients treated with botulinum toxin. AB - AIMS: To evaluate long-term efficacy of intrasphincteric injection of botulinum toxin in untreated achalasia patients; to analyse whether age can be a predictor of response; and to verify whether any objective measurements correlate with symptom relief MATERIALS AND METHODS: A total of 37 patients (mean age 61.4+/ 17.5 years) were enrolled, all of whom injected endoscopically with 100 U of botulinum toxin. Symptom score, oesophageal manometry and oesophageal radionuclide emptying were assessed prior to treatment and 4 weeks, 3 months and 1 year after botulinum toxin. In the case of failure or relapse (symptom score >2), treatment was repeated. RESULTS: All but 6 patients (83.7%) were in clinical remission one month after botulinum toxin. At 12 months, mean symptom score was 0.9+/-0.5 (p<0.05 vs basal); mean lower oesophageal sphincter pressure was 22.0+/ 6.3 (p<0.05 vs basal), and 10-min radionuclide retention was 14.0%+/-7.2 (p<0.05 vs basal). Of the 35 patients followed, 12 (34.3%) had a relapse and were re treated; 4 out of 12 did not respond after re-treatment. Efficacy of first injection of botulinum toxin lasted for a mean period of 15.6 months (range 2 30). Up to day 31 (83.7%) patients were still in remission. We observed a trend towards a better response to botulinum toxin treatment in patients over 50 years (p=0.053). Moreover no correlation was found between any objective achalasia measurements and symptom relief (r coefficient between 0.1 and 0.5) CONCLUSIONS: Results show that: 1) one or two intrasphincteric injections of botulinum toxin result in clinical and objective improvement in about 84% of achalasia patients and are not associated with serious side-effects; 2) patients over 50 years showed better benefit than younger patients; 3) no correlation was found between any objective measurements and symptom relief. PMID- 11926553 TI - Comparison of two new rapid serology tests for diagnosis of Helicobacter pylori infection in Chinese patients. AB - BACKGROUND: Rapid serology test is a simple and convenient way for diagnosing Helicobacter pylori infection. However performances of these tests are usually less satisfactory than expected, particularly in developing countries. AIM: To evaluate the performances of two newly developed rapid serology tests for Helicobacter pylori infection. PATIENTS: Consecutive Chinese dyspeptic patients undergoing upper gastrointestinal endoscopy. METHODS: Gastric biopsies were obtained from antrum and corpus for rapid urease test and histological examination. Diagnosis of Helicobacter pylori infection was based on two or more positive results in rapid urease test, histology and [13C] urea breath test. Patients' sera were tested against two rapid serology tests: ASSURE Hp Rapid Test (Genelabs Diagnostics, Singapore) and SureStep (Applied Biotech, San Diego, CA, USA). RESULTS: A total of 148 patients were evaluated and Helicobacter pylori infection was diagnosed in 78 (53%) patients by gold standard. The sensitivities of ASSURE Hp and SureStep were, respectively, 94% and 71% (p=0.0003). Specificities of the two test kits were both 90%. The overall accuracy of ASSURE Hp was significantly higher than SureStep (92% versus 80%, p=0.004). CONCLUSION: Both rapid serology tests appear to be specific in diagnosing Helicobacter pylori infection in the Chinese populations. However the ASSURE Hp test is more sensitive and accurate than the SureStep test. PMID- 11926554 TI - Evaluation of coeliac disease serological markers in Down syndrome patients. AB - BACKGROUND: The increased incidence of coeliac disease in patients with Down syndrome makes screening of coeliac disease in this population advisable. AIM: Evaluation of efficiency of different serological markers to detect coeliac disease in Down syndrome patients. PATIENTS: A total of 56 Down syndrome patients (aged: 1-17 years) were included in study. METHODS: Patients were evaluated for both IgG and IgA anti-transglutaminase antibodies and for anti-gliadin IgA and IgG antibodies using either purified omega-gliadin, wheat ethanol extract or commercial gliadin. Patients who had at least one positive result were evaluated for antiendomysium antibodies using either monkey oesophagus or human umbilical cord by indirect immunofluorescence. Coeliac disease was diagnosed by typical histological changes on duodenal mucosa. RESULTS: Increased levels of at least one anti-gliadin IgA and IgG antibody marker were found in 27 out of 56 cases (26 for IgG and 9 for IgA). 11/56 were positive for IgG anti-transglutaminase antibodies and two of them were also positive for IgA anti-transglutaminase antibodies and anti-endomysium antibodies. These two patients were finally diagnosed as coeliacs. Gliadin antigenic fractions employed produced differences in the performance of the anti-gliadin IgA and IgG antibody test. The use of commercial gliadin or wheat ethanol extract showed low sensitivity in IgA anti gliadin IgA and IgG antibody determination, whereas good sensitivity and specificity were observed with omega-gliadins. IgG anti-transglutaminase antibodies showed a high proportion of false positive results (9 out of 56), whereas anti-endomysium antibodies and IgA anti-transglutaminase antibodies presented an excellent correlation with presence of active coeliac disease. CONCLUSIONS: Two out of 56 Down syndrome patients were diagnosed as coeliacs, corresponding to an incidence of 3.6%. The use of omega-gliadin presented the best efficiency in anti-gliadin IgA and IgG antibody determination whereas IgA anti-transglutaminase antibodies and anti-endomysium antibody determination showed an absolute correlation with presence of active coeliac disease. PMID- 11926555 TI - Use of arterial conduit as an alternative technique in arterial revascularization during orthotopic liver transplantation. AB - BACKGROUND: The risk of hepatic artery thrombosis after orthotopic liver transplantation is higher in cases of poor hepatic arterial inflow, small or anomalous recipient hepatic arteries, unsafe native hepatic arteries. AIMS: To assess the use of arterial conduits as alternative technique for graft revascularization. PATIENTS: At the Liver Transplant Center of the "S. Giovanni Battista" Hospital in Torino, a review has been made of 600 consecutive orthotopic liver transplantations in 545 adult patients from 1990 to 1999. METHODS: In 95 orthotopic liver transplantations (15.8%) in 88 patients, the graft was supplied by infrarenal conduit, while in 505 orthotopic liver transplantations (84.2%) in 457 patients, a direct anastomosis was used. RESULTS AND CONCLUSIONS: The overall incidence of hepatic artery thrombosis in our series was 3.5% (21/600): 5.3% (5/91) for conduits and 3.2% (16/505) for standard technique (p=ns, chi2 test). The actuarial 5-year graft survival was 67.7% for conduits and 68.6% for the standard technique; p (log rank): ns. The iliac prosthesis torsion was the only complication related to the use of infrarenal iliac conduit. The arterial conduit, performed with donor iliac artery, is an effective and safe revascularization technique in patients at high risk of arterial thrombosis. PMID- 11926556 TI - Evaluation of lanreotide effects on human exocrine pancreatic secretion after a single dose: preliminary study. AB - BACKGROUND: Total parenteral nutrition and somatostatin or analogues represent a consolidated therapeutic approach for external fistulas, a frequent complication of major pancreatic surgery. AIMS: To establish the effects of the somatostatin analogue lanreotide on exocrine pancreatic secretion. METHODS: Eight patients, undergoing pancreaticoduodenectomy for malignancy, were enrolled in the trial. The volume and composition of pancreatic secretion were evaluated after one single subcutaneous injection of lanreotide 0.5 mg or placebo in a randomised, double-blind cross-over trial. RESULTS: In the seven patients completing the study, the 24-h output volume was 208.6+/-41.3 and 253.9+/-72.4 ml after lanreotide and placebo, respectively. During the first 6 hours values were 48.1+/ 14.7 and 77.6+/-21.4 ml (p=0. 02). No significant difference between treatments was detected in the qualitative composition of 24-h pancreatic secretion, although bicarbonate secretion remained lower after the active drug at all the observation intervals. Peak lanreotide levels were detected 15-30 min after drug injection. Clinical and laboratory tolerability was good. CONCLUSIONS: Lanreotide induced a statistically significant reduction in the output volume with respect to placebo in the first 6 hours after administration, but not thereafter. The present results encourage a new study to be undertaken in a larger sample and with a multiple dosing scheme of treatment. PMID- 11926557 TI - New mechanical puncture videoechoendoscope: one-step transmural drainage of a pseudocyst. AB - A new mechanical puncture video echoendoscope (GF-UMD-240P 270 degrees image field parallel to the endoscope axis) has been used for puncture and drainage of a symptomatic pancreatic pseudocyst. It is equipped with a 2.8 mm working channel and an elevator allowing single step drainage with passage of a 7F nasocystic catheter. PMID- 11926558 TI - Diminutive polyps of large bowel should be an early target for endoscopic treatment. AB - BACKGROUND AND AIMS: Aim of the present study is to ascertain the importance of diminutive colorectal polyps and define the need for removal according to their characteristics and malignant potential. PATIENTS AND METHODS: A total of 4,723 patients who underwent colonoscopy were evaluated and 624 patients with 826 polyps were recorded. There were 352 patients with 443 diminutive polyps, studied according to their distribution. Of these, 371 were removed, histologically examined and correlated to patient characteristics and occurrence of synchronous neoplasms. RESULTS: Of the right colon polyps, 81/115 were diminutive, versus 362/711 of the left colon (p<0.0001). Adenomas were more common in patients over 50 years of age, (p<0.0001). In all colonic segments, diminutive adenomas prevailed over hyperplastic polyps, whereas the proportion of diminutive adenomas predominated in the right colon (p=0.0015). Adenomas were classified as tubular 39%, tubulovillous 55.7% and villous 5.3%. The degree of dysplasia was mild in 45.5%, moderate in 51% and severe in 3.5%. The prevalence of synchronous neoplasms was 37.4%. They were more frequently found in males over 50 years of age and in patients with diminutive adenomas compared to those with diminutive hyperplastic polyps (p=0.0078). CONCLUSIONS: The majority of right colon polyps are diminutive. The proportion of diminutive adenomas is higher in patients over 50 years and in the right vs left colon. Diminutive polyps should be removed taking into account the high prevalence of adenomas with a villous component and their significant degree of dysplasia. PMID- 11926559 TI - Upper gastrointestinal bleeding due to duodenal metastasis from primary lung carcinoma. AB - Metastatic dissemination of lung carcinoma may involve virtually all organs, however, symptomatic spread to the small bowel is a rare occurrence. Herein, we report a case of duodenal metastatic involvement leading to severe upper gastrointestinal bleeding in the absence of symptoms referable to the primary tumour. PMID- 11926560 TI - Role of spleen enlargement in cirrhosis with portal hypertension. AB - The possible relationships between splenomegaly and portal hypertension have been analysed in patients with cirrhosis. In this condition, splenomegaly is not only caused by portal congestion, but it is mainly due to tissue hyperplasia and fibrosis. The increase in spleen size is followed by an increase in splenic blood flow, which participates in portal hypertension actively congesting the portal system. PMID- 11926561 TI - Is insulin resistance a pathogenic co-factor in hepatitis C virus-related disease and hepatocellular carcinoma? PMID- 11926562 TI - Hepatic-intestinal function after total gastrectomy. PMID- 11926563 TI - 3020insC mutation within the NOD2 gene in Crohn's disease: frequency and association with clinical pattern in an Italian population. PMID- 11926564 TI - Selective cyclooxygenase-2 inhibitors: after the smoke has cleared. AB - An enormous amount of fanfare and marketing preceded the introduction of selective inhibitors of cyclooxygenase-2 to the marketplace. These drugs were purported to offer equivalent anti-inflammatory and analgesic effects to conventional non-steroidal anti-inflammatory drugs without causing gastrointestinal injury. Now that there is considerable clinical experience with four drugs of this class having been available for at least two years, it is worthwhile re-visiting some of the original claims to determine whether selective cyclooxygenase-2 inhibitors have thus far lived up to their promise. In short, selective cyclooxygenase-2 inhibitors have proven to be somewhat safer in terms of gastrointestinal toxicity, than some (but not all) conventional non-steroidal antiinflammatory drugs. However their efficacy of the selective cyclooxygenase inhibitors has not always matched that of the conventional nonsteroidal anti inflammatory drugs and there are significant safety concerns with some of the new drugs that deserve very careful consideration. PMID- 11926565 TI - Proton pump inhibitors or surgery for gastro-oesophageal reflux disease. PMID- 11926566 TI - Botulinum toxin for achalasia: are we asking the right questions? PMID- 11926567 TI - Laparoscopic fundoplication versus lansoprazole for gastro-oesophageal reflux disease. A pH-metric comparison. AB - BACKGROUND: Treatment strategies that abolish abnormal reflux could prevent long term complications of gastro-oesophageal reflux disease. AIMS: To compare the efficacy of laparoscopic fundoplication and lansoprazole in abolishing abnormal reflux in patients with gastro-oesophageal reflux disease. PATIENTS: Study population comprised 130 patients referred for possible antireflux surgery and with heartburn as the dominant symptom. METHODS: After oesophageal manometric and pH-metric evaluation and detailed information 55 patients asked to undergo laparoscopic antireflux surgery while 75 chose a medical treatment regimen based on lansoprazole. Treatment efficacy was assessed by ambulatory oesophageal pH monitoring. RESULTS: All 55 patients who underwent fundoplication became free of heartburn: oesophageal pH-monitoring gave normal results in 85%. In patients treated with lansoprazole, at individualized daily dosages titrated to abolish both heartburn and abnormal acid reflux, normal pH-metric results were obtained in 96% of cases (p<0.05 vs surgically treated patients). CONCLUSIONS: Lansoprazole at individualized dosages was significantly more effective than laparoscopic fundoplication, in the short-term, in abolishing abnormal reflux in gastro-oesophageal reflux disease patients. PMID- 11926568 TI - Coeliac disease and secondary autoimmunity. PMID- 11926569 TI - Regression of duodenal gastric metaplasia in Helicobacter pylori positive patients with duodenal ulcer disease. AB - BACKGROUND: It is unclear whether the extent of duodenal gastric metaplasia is due to Helicobacter pylori and/or acid. AIMS: To investigate the role of Helicobacter pylori eradication in the regression of duodenal gastric metaplasia in patients with duodenal ulcer maintained in acid suppression conditions. METHODS: . Duodenal (anterior, superior inferior walls of first part of duodenum) and gastric antrum biopsies were obtained from 44 Helicobacter pylori positive duodenal ulcer patients. Helicobacter pylori infection was diagnosed by rapid urease test, histology and 13C-Urea Breath Test. Patients were treated with 20 mg omeprazole tid associated with 250 mg clarithromycin and 500 mg amoxycillin four times daily for 10 days and maintained with 20 mg omeprazole daily for 18 weeks. Control endoscopies were performed at 6 and 18 weeks after beginning treatment. RESULTS: Duodenal gastric metaplasia regression was observed in all (32/32) patients in whom Helicobacter pylori was eradicated, but in only 3 out of 6 patients in whom eradication was not achieved (p<0. 001). CONCLUSIONS: . The present results suggest that Helicobacter pylori eradication associated with prolonged acid suppression may represent a good therapeutic strategy to achieve duodenal gastric metaplasia regression and highlight the combined role of acid and Helicobacter pylori in the pathogenesis of duodenal gastric metaplasia. PMID- 11926570 TI - Voyage of Helicobacter pylori in human stomach: odyssey of a bacterium. PMID- 11926571 TI - Treatment of severe Crohn's disease using antimycobacterial triple therapy- approaching a cure? AB - BACKGROUND: Mycobacterium avium subspecies paratuberculosis is probably the best candidate for a microbial cause of Crohn's disease although arguments to the contrary can be equally convincing. Growing evidence suggests that prolonged antimycobacterial combination therapy can improve Crohn's disease in some patients. AIM: To report long-term observations in patients with severe Crohn's disease treated with triple macrolide-based antimycobacterial therapy. PATIENTS: A series of 12 patients (7 male, 5 female; aged 15-42 years) with severe, obstructive or penetrating Crohn's disease were recruited. METHODS: Patients failing maximal therapy were commenced prospectively on a combination of rifabutin (450 mg/d), clarithromycin (750 mg/d) and clofazimine (2 mg/kg/d). Progress was monitored through colonoscopy, histology, clinical response and Harvey-Bradshaw activity index. RESULTS: Follow-up data were available for up to 54 months of therapy Six out of 12 patients experienced a full response to the antiMycobacterium avium subspecies paratuberculosis combination achieving complete clinical, colonoscopic and histologic remission of Crohn's disease. Four of these patients were able to cease treatment after 24-46 months, 3 of whom remained in total remission without treatment for up to 26 months and one patient relapsed after six months off treatment. A partial response to the anti Mycobacterium avium subspecies paratuberculosis combination was seen in 2 patients showing complete clinical remission with mild histologic inflammation. Return to normal of terminal ileal strictures occurred in 5 patients. Harvey Bradshaw activity index in patients showing a full or partial response to therapy fell from an initial 13.4 +/- 1. 91 to 0. 5 +/- 0. 47 [n = 8, p < 0. 001) after 52-54 months. CONCLUSIONS: Reversal of severe Crohn's disease has been achieved in 6/12 patients using prolonged combination anti-Mycobacterium avium subspecies paratuberculosis therapy alone. Three patients remain in long-term remission with no detectable Crohn's disease off all therapy These results support a causal role for Mycobacterium avium subspecies paratuberculosis in Crohn's disease while also suggesting that a cure may become possible. PMID- 11926572 TI - High frequency of anti-endomysial reactivity in candidates to heart transplant. AB - BACKGROUND: A possible link between coeliac disease and dilated cardiomyopathy has recently been suggested. AIMS: . To assess the frequency of anti-endomysial antibodies, the marker for coeliac disease, in patients with different forms of heart failure, and to establish the clinical features of those endomysial antibody positive. SUBJECTS AND METHODS: . A total of 642 consecutive patients entering the waiting list for heart transplantation from 1995 through 1997 were studied. The prevalence of endomysial IgA antibodies, determined by indirect immunofluorescence, was compared to that observed in three surveys conducted in the Italian general population. RESULTS: Of the 642 patients, 12 (1.9%; 95% confidence interval 0.97-3.2) resulted endomysial antibody positive, versus 34/9,720 healthy controls (0.35%; 95% confidence interval, 0.23-0.47), accounting for a relative risk of 5.3 (95% confidence interval, 2.8-10.3). Anti-endomysial antibodies were found in 6/275 patients with dilated cardiomyopathy and 6/367 with other forms of heart failure (2.2% versus 1.6%; 95% confidence interval 0.8 4.7 and 0.6-3.5), with no statistical difference. The 12 endomysial antibody positive patients were leaner (body mass index, 22.0 +/- 1.9 vs 24.2 +/- 3. 1, p<0. 05) than 36 seronegative patients matched for baseline demographics and aetiology of cardiomyopathy No differences were observed as regards clinical, biochemical and echocardiographic features, mortality in waiting list and 2-year post-transplant survival. CONCLUSIONS: Patients with end-stage heart failure are at increased risk for coeliac disease as compared to the general population. PMID- 11926573 TI - Severe attack of ulcerative colitis in children: retrospective clinical survey. AB - AIM: To obtain clinical data concerning severe attacks of ulcerative colitis in children. PATIENTS AND METHODS: A retrospective chart review of 37 children with ulcerative colitis was carried out in order to assess the prevalence, risk factors, timing of presentation, and outcome of severe attacks of ulcerative colitis. RESULTS: A total of 20 severe attacks occurred in 15 out of the 37 patients. No difference in the occurrence of severe attacks was detected in relation to age or disease extent at diagnosis. The mean interval between disease diagnosis and a severe attack was 9.1 months (range 0-30). Of the 20 severe attacks, 11 were resolved with medical treatment in a mean time of 1 1 +/- 4.6 days while 9 out of 20 needed urgent surgery in a mean time of 7.4 +/- 4.8 days. Of 10 out of the 15 patients who recovered from the first attack 4 required colectomy after a mean time of 6.7 months, another 4 are still in remission at a mean period of 40.7 months, one needed elective surgery after 25 months and one was lost to follow-up. CONCLUSIONS: Severe attacks of ulcerative colitis had a high prevalence rate [40%); age and disease extent at presentation were not predictors of their occurrence. Approximately half the attacks resolved with medical treatment alone, while the other half required emergency surgery. After successful medical treatment of the first attack, 40% of children maintained long term remission, while 40% required early colectomy PMID- 11926575 TI - Prevention and chemoprevention of colorectal neoplasms. AB - Main purpose of the review is to analyse the impact of the current approaches for colorectal cancer prevention, including chemoprevention. Available evidence does not support the contention that a more appropriate diet can be of great help in the prevention of these neoplasms, either because the scientific evidence is poor and highly controversial, or because changes in diet are difficult to implement, at least in many Western countries. Similarly, a preventive approach based on the modification of lifestyle remains improbable, either in the short --or in the long period of time. Secondary prevention--i.e., the systematic removal of adenomatous polyps--can hardly be applied in the general population, with the exception of individuals at risk because members of families with Adenomatosis coli or Lynch syndrome, or affected by inflammatory bowel diseases. Finally, chemoprevention (i.e., the attempt to prevent tumour development through the administration of drugs or natural compounds that interfere with various phases of carcinogenesis) is still in its infancy Though attractive, this approach requires well-designed studies which should be carried out for years before being evaluated and interpreted; so far most of these investigations gave inconsistent or controversial results. In conclusion, both primary and secondary prevention of colorectal malignancies appear difficult to apply in the general population, and chemoprevention is still at the beginning of a (presumably] long story. The final impression is that notwithstanding the remarkable advancements made in the last two decades in colorectal cancer research, the practical application of these new concepts remains difficult. PMID- 11926574 TI - Are hepatitis G virus and TT virus involved in cryptogenic chronic liver disease? AB - BACKGROUND: Hepatitis G virus can cause chronic infection in man but the role of this agent in chronic liver disease is poorly understood. Little is known about the relation of another newly discovered agent, the TT virus, with chronic liver disease. AIM: To investigate the rate of infection with hepatitis G virus and TT virus in patients with cryptogenic chronic liver disease. PATIENTS: A total of 23 subjects with chronically raised alanine transaminase and a liver biopsy in whom all known causes of liver disease had been excluded, and 40 subjects with hepatitis C virus-related chronic liver disease. METHODS: Evaluation of anti hepatitis G virus by enzyme immunoassay. Hepatitis G virus-RNA by polymerase chain reaction with primers from the 5' NC and NS5a regions. TT virus-DNA by nested polymerase chain reaction with primers from the ORF1 region. Results. Hepatitis G virus-RNA was detected in 4 out of 23 patients with cryptogenic chronic hepatitis and in 6 out of 40 with hepatitis C virus chronic hepatitis (17.4% vs 15% p=ns). At least one marker of hepatitis G virus infection (hepatitis G virus-RNA and/or anti-hepatitis G virus, mostly mutually exclusive) was present in 6 out of 23 patients with cryptogenic hepatitis and 16 out of 40 with hepatitis C virus liver disease (26. 1% vs 40% p=ns). T virus-DNA was present in serum in 3 subjects, 1 with cryptogenic and 2 with hepatitis C virus related chronic liver disease. Demographic and clinical features, including stage and grade of liver histology, were comparable between hepatitis G virus-infected and uninfected subjects. Severe liver damage [chronic hepatitis with fibrosis or cirrhosis) were significantly more frequent in subjects with hepatitis C virus liver disease. CONCLUSIONS: In Southern Italy, hepatitis G virus infection is widespread among patients with chronic hepatitis, independently of parenteral risk factors. Its frequency in subjects with cryptogenic liver disease parallels that observed in hepatitis C virus chronic liver disease, thus ruling out an aetiologic role of hepatitis G virus. TT virus infection is uncommon in patients with cryptogenic or hepatitis C virus-related liver disease who do not have a history of parenteral exposure. PMID- 11926577 TI - Therapy in inflammatory bowel disease: a double-edged sword. PMID- 11926576 TI - Treatment of Helicobacter pylori infection. Indications and regimens: an update. AB - The management of Helicobacter pylori infection is still surrounded by controversy and uncertainties. Indications and correct application of current regimens for Helicobacter pylori infection are still considered a matter of debate. Regarding indications, only peptic ulcer and mucosa associated lymphoid tissue lymphoma are considered clear indications for treatment. In other conditions, such as atrophic gastritis, post gastric cancer resection, first degree relatives of gastric cancer patients, dyspeptic patients, patients with gastro-oesophageal reflux disease and non-steroidal anti-inflammatory drug users, the value of Helicobacter pylori eradication is still controversial. The regimens for first-line and second-line treatment of Helicobacter pylori infection have been recommended by the Maastricht 2 Consensus Report. Although all the treatments are considered to be effective, physicians still do not agree on what first-line regimen should be used. Furthermore, a consensus on the duration of the antibiotic treatment is still lacking, although Maastricht guidelines for treatment of Helicobacter pylori infection recommend a one-week therapy. Also regimens, as a third-line treatment, and methods to improve compliance and clinical outcome are still a matter of debate. All these points will be considered in the present review PMID- 11926578 TI - Ogilvie's syndrome in patient with multiple myeloma. PMID- 11926579 TI - Helicobacter pylori infection and hepatic encephalopathy. PMID- 11926580 TI - Treatment with drugs active against Mycobacterium avium subspecies paratuberculosis can heal Crohn's disease: more evidence for a neglected public health tragedy. PMID- 11926581 TI - Computed tomography characteristics of non-syndromic craniofacial fibrous dysplasia. AB - BACKGROUND: Fibrous dysplasia is a benign fibro-osseous tumor of bones commonly involving the craniofacial region. Computed tomography (CT) imaging study of the disease is useful for evaluation and treatment planning. However, few studies have evaluated such large patient series. METHODS: A total of 46 patients with complete medical records and CT images was included in this study. All of these patients were non-syndromic, had fibrous dysplasia involving only the craniofacial region, and had no skin pigmentation or other evidence of endocrine problems. Data analyses were performed on the clinical manifestations, time of onset, signs and symptoms, involvement of cranial and facial bones, and CT appearance of the tumors in this patient group. RESULTS: Painless swelling was the chief clinical problem in 78% of patients, followes by dental malocclusion in 22%. Onset of the disease was reported to have occurred before 6 years of age in 34%, between 6 and 10 years in 27%, and older than 10 years in 39% of patients. Extreme timings such as onset at infancy or older than 20 years of age were also noted. The average number of bones involved was 3.2 bones per patient. Involvement of more than one craniofacial bone occurred in 70% of patients. The maxilla, orbital, and frontal bones were most commonly involved. CT images appeared sclerotic or homogenous in 34%, mixed white and dark or heterogenous in 55%, and cystic in 11%. A correlation between the age of onset of the disease and the number of bones involved was not observed. CONCLUSIONS: Findings of this study demonstrate that craniofacial fibrous dysplasia displays a wide spectrum of clinical behaviors. CT imaging generally revealed extensive involvement of the tumor in the craniofacial region. PMID- 11926582 TI - Sub-tenon anesthesia for segmental scleral buckling and assessment of postoperative pain. AB - BACKGROUND: To evaluate the safety and efficacy of sub-Tenon anesthesia for segmental scleral buckling. METHODS: Thirty-two patients diagnosed with rhegmatogenous retinal detachment were treated with segmental scleral buckling under sub-Tenon anesthesia. After topical anesthesia, a buttonhole was made through the conjuntiva and Tenon's capsule 4 mm posterior to the limbus. Four milliliters of anesthetic solution was then delivered into the posterior sub Tenon space using a blunt cannula. The buckling procedure was done immediately after the completion of anesthesia. We evaluated akinesia and recorded the pain with a visual analogue scale after surgery. RESULTS: There were no anesthesia related complications. Twenty-two patients (69%) reported no pain during surgery. Nine patients (28%) felt pain during surgery. However, the pain was tolerable and the surgeries were finished smoothly with or without a supplemental anesthetic solution. One patient (3%) experienced uncomfortable pain and needed an additional retrobulbar block. Five patients (16%) retained complete eye movement 5 min after anesthesia, and only 4 patients (13%) experienced total akinesia. At the end of the surgery, 16 patients (50%) had total akinesia and 2 patients (6%) retained complete eye movement. CONCLUSIONS: Sub-Tenon anesthesia is efficient and safe in segmental scleral buckling. It can prevent the complications of peribulbar or retrobulbar anesthesia and is a good alternative to both methods of anesthesia, especially in highly myopic eyes. PMID- 11926583 TI - Spleen-preserving distal pancreatectomy without division of splenic artery and vein as a procedure for benign distal pancreatic lesion. AB - BACKGROUND: To assess the safety and the clinical outcome of distal pancreatectomy, with preservation of the spleen as well as splenic artery and vein, for benign distal pancreatic lesions. METHODS: Five consecutive patients with benign distal pancreatic lesions (3 with insulinoma, 1 with non-functioning islet cell tumor and 1 with serous cystadenoma) underwent spleen-preserving distal pancreatectomy with conservation of the splenic artery and vein. Prograde distal pancreatectomy was performed for 4 patients and retrograde pancreatectomy for the other. The operative time, blood loss due to surgery, length of post operative hospitalization and post-operative complications were analyzed and evaluated. RESULTS: Surgery was successful for all 5 patients. Whilst 2 of the patients revealed major medical disease, no post-surgical complications were experienced by any of the 5 patients. The mean operative time, extent of blood loss, and postoperative hospital stay were, respectively, 238 minutes (range 175 270), 170 ml (range 50-300), and 8.4 days (range 6 - 15). CONCLUSION: From our experience, spleen-preserving distal pancreatectomy can be safely performed with the conservation of the splenic artery and vein. Our result revealed that this well-known procedure can be improved in terms of blood loss, surgical duration and length of hospital stay. We believe that this procedure should be performed for benign lesions of the distal pancreas whenever indicated and possible. PMID- 11926584 TI - Findings of anthropometric and laboratory data from adult health screening under the National Health Insurance plan in Taiwan. AB - BACKGROUND: From April of 1996, all adults over 40 years old insured by the National Health Insurance (NHI) plan of Taiwan are eligible for a physical check up once a year by most NHI-contracted family physicians. This study provides a benchmark data of their current physical health. METHODS: In total, 9,016 adult health examination reports were systematically sampled from 692,311 claims from April to September 1996. A subject profile included basic personal information, a physical examination, and routine blood and urine laboratory tests. The reliability of the data was verified by comparing the results of anthropometrical measures to others obtained from a nationwide nutrition survey. RESULTS: The mean height was 165 +/- 6.3 cm for men and 154 +/- 5.6 cm for women. Totally 54.3% of the people sampled were overweight, especially younger adults. The average systolic/diastolic blood pressure was 129+/- 19.9/80.9+/- 11.7 mmHg for men and 127.2 +/ -21.2/78.9 +/- 12.0 mmHg for women. There were 17.6% with high blood sugar, while only 8.0% were self-reported as diabetes mellitus (DM) sufferers. Women over 65 had higher cholesterol levels (>6.24 mmol/L) than did their younger peers and men. Men's blood triglyceride levels decreased from 1.74 to 1.28 g/L with age, while women's increased from 1.04 to 1.55 g/L with age. CONCLUSIONS: Taiwanese adults over 40 are 2-3 kg heavier and 2 cm taller than their counterparts 10 years ago, but may not be healthier. Future study may need to focus upon cost-effectiveness of this nationwide adult health screening program. PMID- 11926585 TI - Endoscopic resection of a large colonic leiomyoma. AB - A 48-year old man suffered from intermittent hematochezia and abdominal cramping over a period of four weeks. A colonoscopy revealed a 4.5 cm semi-pedunculated tumor in the transverse colon. Using a 2-channel colonoscope, the tumor was successfully removed with an electro-surgical snare after normal saline submucosal injection. Microscopic examination revealed it to be a smooth muscle tumor without mitosis. The patient recovered well, and did not have a residual tumor in a follow-up colonoscopy. We found no reports of a colonic leiomyoma larger than 3 cm which was completely removed by a colonoscope. According to the case presented here, intraluminal colonic leiomyoma can be completely resected with skillful manipulation of a colonoscope, even if the tumor is as large as 4.5 cm. Successful endoscopic polypectomy of colonic leiomyoma reduces the cost of treatment and eliminates unnecessary surgery. PMID- 11926586 TI - Bilateral orbital Kimura's disease in a young Asian man. AB - A 16-year-old Asian man adolescent presented with bilateral eyelid swelling with multiple palpable mass lesions, which waned after treatment with corticosteroids but waxed after medications were discontinued, for about 1 year. He was otherwise healthy except that bilateral postauricular lymphadenopathy had developed for about 9 years. Laboratory study revealed peripheral eosinophilia with an elevated IgE level. The tumor masses of the left orbit were completely excised through an incision similar to that used in blepharoplasty, which gave good cosmetic results. Postoperative computed tomography scan showed no residual tumor mass in the left orbit but contralateral homogeneous soft-tissue mass lesions around the lacrimal gland and extending deep into the orbit, between the superior and lateral rectus muscles. Pathology reported numerous lymphoid follicles with active germinal centers and extensive lymphocyte and eosinophil infiltration, which characterize Kimura's disease (KD). The tumor mass in the right orbit was also excised during a second elective surgery 10 days later. No evidence of recurrence was noted after follow-up for 7 months. KD of the orbit is rare and usually occurs in middle-aged to elderly Asian men but can also be present in young adolescent. Complete excision with simultaneous blepharoplasty gives satisfactory cosmetic results. PMID- 11926588 TI - Eosinophilic gastroenteritis involving the distal small intestine and proximal colon. AB - Eosinophilic gastroenteritis (EG) is an unusual disorder. It is characterized by eosinophil infiltration of the gut wall histologically and is manifested by gastrointestinal (GI) symptoms clinically. This disease entity preferentially affects the stomach and proximal small intestine. Mucosal layer disease is the most common form of this uncommon disease. We present a case of EG with transmural distal small intestinal and proximal colonic involvement whose clinical symptoms included watery diarrhea, abdominal pain, and body weight loss. Colonoscopy showed non-specific colitis in the proximal colon. Small bowel series showed diffuse jejunal dilatation with wall thickening and rigidity. Abdominal computed tomography also showed a thickened bowel wall with partial ileus and ascites. Diagnosis was established through endoscopic biopsy and ascites paracentesis, while at the same time excluding the possibility of parasite infection. Treatment with prednisolone produced a dramatic response. A high index of suspicion in cases of peripheral eosinophilia with concomitant GI symptoms is needed for the early diagnosis of this uncommon disease. PMID- 11926587 TI - An unusual case of dual coronary artery fistulas to main pulmonary artery. AB - Coronary artery fistula is an anomaly in which a coronary artery directly connects to a cardiac chamber or great vessel. Its incidence is around 0.1 to 1% in the adult population. Dual coronary artery fistulas are far less common and their incidence is estimated to be around 5% in patients with this anomaly. Closure of the fistulas is indicated in patients with myocardial ischemia, large left to right shunt, congestive heart failure or other complications. Herein, we report a 64 year-old man with dual coronary artery fistulas presenting with exertional chest pain. The fistulas were initially suspected on transesophageal echocardiogram because of abnormal flow with a mosaic pattern between the left anterior descending and main pulmonary arteries. Selective coronary angiogram confirmed the diagnosis and revealed fistulous connections from the proximal left anterior descending and ostial right coronary arteries to the main pulmonary artery. The patient became symptom-free after surgical closure of the fistulas. PMID- 11926589 TI - Minimally invasive procedure for ventriculoatrial shunt-combining a percutaneous approach with real-time transesophageal echocardiogram monitoring: report of six cases. AB - To make the ventriculo atrial (VA) shunt procedure less invasive and avoid atrial catheter malpositioning, we combined a percutaneous approach with transesophageal echocardiogram (TEE) to monitor the atrial catheter tip. From July 1995 to October 1996, we performed 6 VA shunt procedures by combining these 2 techniques. Except for one patient who had a concomitant Port-A catheter, which obscured the detection of TEE in the procedure, all other atrial catheters achieved ideal positioning and good function. The mean operation time was shorter, and the complications were fewer than those using the conventional procedure in our hospital. In addition to this particular aspect, by combining these 2 techniques, the VA shunt procedure can be performed much less invasively, but more accurately, quickly, and safely. PMID- 11926590 TI - Translation and validation assessment of the Chinese version of the chronic sinusitis survey. AB - BACKGROUND: The Chronic Sinusitis Survey (CSS) is a valid, disease-specific measure for evaluating the health status and treatment effectiveness of adults with chronic rhinosinusitis (CRS). In this study, we developed a Chinese version of the CSS (CCSS) which provides the psychometric properties of the Chinese CSS. METHODS: The CSS was translated into Chinese using a parallel model. The CCSS was administered to 198 patients in a prospective manner, and was validated in order to establish its reliability and validity. RESULTS: The CCSS demonstrated good test-retest reliability (correlation coefficient = 0.6-0.89, p=0.0001) and internal consistency (Cronbach's alpha =0.76). The CCSS results were significantly correlated with bodily pain (BP), general health (GH), role emotional (RE), and mental health (MH) subscales of the Chinese (Taiwan) version of the generic 36-item Short-Form Health Survey (TSF-36). The standardized response mean for the CCSS total score was 0.75, indicating good sensitivity to clinical change. CONCLUSIONS: This validation study demonstrates that the performance characteristics of the CCSS meet the criteria for a valid measure. The CCSS is a valid tool to evaluate adults with CRS among Mandarin-speaking populations. PMID- 11926592 TI - Effect of Tityus serrulatus venom on cytokine production and the activity of murine macrophages. AB - THE purpose of this study was to investigate the effects of Tityus serrulatus venom (TSV) on murine peritoneal macrophages evaluated in terms of activation. The effects of crude TSV were analysed by detection of cytokines, oxygen intermediate metabolites (H2O2) and nitric oxide (NO) in supernatants of peritoneal macrophages. Several functional bioassays were employed including an in vitro model for envenomating: cytotoxicity of TSV was assessed using the lyses percentage. Tumor necrosis factor (TNF) activity was assayed by measuring its cytotoxic activity on L-929 cells, and interleukin-6 (IL-6) and interferon-gamma (IFN-gamma) were assayed by enzyme-linked immunosorbent assay, whereas NO levels were detected by Griess colorimetric reactions in culture supernatant of macrophages incubated with TSV and subsequently exposed to either lipopolysaccharide or IFN-gamma. Incubation of macrophages with TSV increased production of IL-6 and IFN-gamma in a dose-dependent manner. TNF production was not detected in supernatants treated with TSV at any concentration. The increase in IL-6 secretion was not associated with concentration-dependent cytoxicity of TSV on these cells. These data suggest that the cytotoxicity does not appear to be the main cause of an increased cytokine production by these cells. Although NO is an important effector molecule in macrophage microbicidal activity, the inducing potential of the test compounds for its release was found to be very moderate, ranging from 125 to 800 mM. Interestingly, NO levels of peritoneal macrophages were increased after IFN-gamma. Moreover, NO production had an apparent effect on macrophage activity. The results obtained here also shown that the TSV induces an important elevation in H2O2 release. These results combined with NO production suggest that TSV possesses significant immunomodulatory activities capable of stimulating immune functions in vitro. PMID- 11926591 TI - The expression of COX-2 in VEGF-treated endothelial cells is mediated through protein tyrosine kinase. AB - Cyclooxygenase (COX), existing as the COX-1 and COX-2 isoforms, converts arachidonic acid to prostaglandin H2, which is then further metabolized to various prostaglandins. Vascular endothelial growth factor (VEGF) has been shown to play important roles in inflammation and is upregulated by the prostaglandin E series through COX-2 in several cell types. Here, we have investigated the effects of VEGF on the COX isoform expressed in human umbilical vein endothelial cells (HUVEC). The signalling mechanism of the COX isoform expressed in endothelial cells activated with VEGF will be also investigated using the tyrosine kinase inhibitor, genistein, and protein kinase C inhibitor, staurosporine. The activity of COX-2 was assessed by measuring the production of 6-keto-prostaglandin F1alpha in the presence of exogenous arachidonic acids (10 microM, 10 min) by enzyme immunoassay. The expression of COX isoform protein was detected by immunoblot using specific antibodies. Untreated HUVEC contained no COX-2 protein. In HUVEC treated with VEGF (0.01-50 ng/ml), COX-2 protein, but not COX-1, and COX activity were increased in a dose-dependent manner. Interestingly, the increased COX-2 protein and activity in response to VEGF (10 ng/ml) was inhibited by the tyrosine kinase inhibitor, genistein (0.05-5 microg/ml), but not by the protein kinase C inhibitor, staurosporine (0.1-10 ng/ml). Thus, the induction of COX-2 by VEGF in endothelial cells was mediated through protein tyrosine kinase, and the uses of specific COX-2 inhibitors in these conditions, in which VEGF was involved, might have a role. PMID- 11926593 TI - Epithelium-dependent effect of L-glutamate on airways: involvement of prostaglandins. AB - We investigated the effect of the excitatory amino acid (EAA) receptor agonists L glutamate, N-methyl-D-aspartate (NMDA), (RS)-a-amino-3-hydroxy-5-methyl-4 isoxazole propionic acid (AMPA) and kainic acid on KCl-induced contractions of rabbit tracheal smooth muscle, as well as the role of epithelium and endogenously produced nitric oxide and prostaglandins on these responses. L-Glutamate decreased KCI-induced contractions up to 30%. This effect was attenuated by epithelium removal, tetrodotoxin, methylene blue and indomethacin but not by NG nitro-L-arginine methyl ester. While NMDA, AMPA and kainic acid had no effect, the combination of NMDA + kainic acid decreased KCI-induced contractions. These results suggest that, in rabbit trachea, L-glutamate has, at least in part, an epithelium-dependent effect mediated via prostaglandin formation and that the EAA receptors involved are non-classical. PMID- 11926594 TI - IL-1 stimulates ceramide accumulation without inducing apoptosis in intestinal epithelial cells. AB - BACKGROUND: In inflammatory bowel disease (IBD), cytokine levels (such as interleukin-1 (IL-1)) are elevated. We have shown previously that IL-1 activates phospholipid signaling pathways in intestinal epithelial cells (EEC), leading to increased ceramide levels. AIM: To determine whether ceramide induces apoptosis in IEC. METHODS: Apoptosis was evaluated by annexin-V binding or Hoechst nuclear staining. Levels of bcl-2, bcl-x, bax, p53 and p21 were determined by Western blotting, and celi cycle analysis was determined by flow cytometry. RESULTS: IL-1 increased ceramide accumulation in a time-dependent and concentration-dependent manner with a peak response at 4 h, with [IL-1] = 30 ng/ml. Neither IL-1 nor ceramide induced apoptosis in EEC, but they increased bcl-2 levels and decreased bax and p21 levels without affecting bcl-x and p53 levels. They also caused a slight but significant increase in the G2/M phase. These data suggest a role for ceramide in IBD and suggest a possible mechanism for the enhanced tumorigenic activity in IBD patients. PMID- 11926596 TI - Azelastine and suplatast shorten the distribution half-life of IgE in rats. AB - We aim to clarify whether suplatast and azelastine (anti-allergic drugs) can shorten the half-life of imnunoglobulin E (IgE) in the circulating blood. Thirty Wistar rats were divided into six groups. Distilled water or anti-allergic drugs were given orally for 6 days after the first sensitization. Two milligrams of monoclonal dinitrophenyl (DNP)-specific rat IgE was administered to the rats, which had been given suplatast or azelastine orally. The level of DNP-specific rat IgE in the serum was estimated by IgE-capture enzyme-linked immunosorbent assay, and the turnover of IgE was analyzed from its pharmacokinetic parameters. The elimination half-life of rat IgE was about 12 h irrespective of the sensitized state. The intercompartmental rate constants (Kct and Ktc) in the suplatast-administered or azelastine-administered group were larger than those of the distilled water-administered group under non-sensitized conditions. These findings suggested that the anti-allergic drugs used in the present study facilitated the excretion of IgE from the circulation in rats. PMID- 11926595 TI - Immunomodulating effects of tofizopam (Grandaxin) and diazepam in vitro. AB - Benzodiazepines (BDZs) are known to act not only in the central nervous system, but on peripheral cells and tissues binding to the peripheral-type benzodiazepine receptors. In the present study, the influence of two different BDZs (diazepam (Dz) and tofizopam (Tof) on several immune functions has been examined in vitro. Some differences between Dz and Tof in their effects on human lymphocyte proliferative response, changes in glucocorticoid-induced suppression of cell proliferation and influence on cytokine production (tumor necrosis factor-alpha (TNF-alpha) and interleukin-2 (IL-2)) have been determined. Dz suppressed mitogen induced peripheral blood mononuclear cell (PBMC) proliferation, enhanced dexamethasone-induced inhibition of PBMC proliferative response, and suppressed lymphocyte production of TNF-alpha and IL-2. Tof usually enhanced PBMC proliferation and IL-2 production in low and moderate doses, but in high doses it suppressed both. Tof in all investigated doses enhanced dexamethasone-induced suppression of lymphocyte proliferation and depressed TNF-alpha production. Thus, both Dz and Tof are shown to have immunomodulating effects in vitro. Tof, opposite to Dz even in the therapeutic doses, is able to enhance in vitro mitogen induced lymphocyte proliferation and IL-2 production. PMID- 11926597 TI - The presence of elafin, SLPI, IL1-RA and STNFalpha RI in head and neck squamous cell carcinomas and their relation to the degree of tumour differentiation. AB - Biopsy samples of head and neck carcinomas were investigated with regard to elafin, secretory leukocyte protease inhibitor (SLPI), interleukin 1-receptor antagonist [(IL)1-RA] and soluble tumour necrosis factor alpha receptor antagonist (STNFalpha RI). SLPI and elafin are serine protease inhibitors produced in the serous cells of the upper respiratory airways and in the keratinocytes, respectively. We have now found the presence of elafin and SLPI in squamous cell carcinomas of the upper respiratory tract (tonsillar, hypopharyngeal, tongue, mouth floor, gingival and laryngeal cancer). Significantly higher amounts of SLPI and elafin are present in well differentiated and moderately differentiated tumours than in poorly differentiated tumours (p < 0.0001 and p < 0.0015). Tumour necrosis factor-alpha and IL-1beta have been shown to stimulate the production of SLPI and elafin. Since these cytokines can both be difficult to detect, we chose to study their inhibitors, STNFalpha RI and IL1-RA, instead. IL1-RA was expressed in highly differentiated tumours as well as in poorly differentiated ones. No significant difference was seen between the groups. STNFalpha RI was only found in very small amounts, sparsely distributed in the tumours, and was not related to the degree of differentiation. PMID- 11926598 TI - Single-use acupuncture needles: scanning electron-microscopy of needle-tips. AB - Unselected, disposable acupuncture needles from various manufacturers and retail suppliers were taken from a pool of donated and bought samples. Three needles of each type were prepared for electron-microscopy. The needle tips were inspected at two magnifications (x39.37 and x612.5): 52 electron-micrographs were taken of 31 individual needles from 11 different types. No needle-tip looked perfect and significant faults were seen in most; some appeared seriously deformed. The faults noted were: scratch marks along or across the needle, metallic scuff, lumps and irregularities in the needle surface, needle-tip stubbed or malformed, and needle point off-centre. An additional test made was to wipe a number of needles firmly on white paper tissue. Some left grey lines, and these were regarded as evidence of metallic or oily residue from the needle surface which could have been deposited in the patient. These unexpected findings, in a variety of popularly used needles from well-respected suppliers, suggest that most manufacturers need to reassess their quality control procedures. PMID- 11926599 TI - Effect of acupuncture on knee function in advanced osteoarthritis of the knee: a prospective, non-randomised controlled study. AB - We report a prospective controlled trial, comparing acupuncture with no treatment, in patients with advanced osteoarthritis of the knee awaiting total knee replacement. Knee function was assessed at the beginning of study and at the end of two months, using four parameters: HSS score, time to walk 50 metres, time to climb 20 steps, and degree of pain. Acupuncture was given at four local points around the knee and at one distal point. The acupuncture group improved in all parameters, whereas the control group deteriorated, a finding that was highly statistically significant (p < 0.0002). Further randomised-controlled trials with longer follow-up are required to confirm these findings. PMID- 11926600 TI - Management of myofascial trigger point pain. AB - Successful management of myofascial trigger point (MTrP) pain depends on the practitioner finding all of the MTrPs from which the pain is emanating, and then deactivating them by one of several currently used methods. These include deeply applied procedures, such as an injection of a local anaesthetic into MTrPs and deep dry needling (DDN), and superficially applied ones, including an injection of saline into the skin and superficial dry needling (SDN) at MTrP sites. Reasons are given for believing that DDN should be employed in cases where there is severe muscle spasm due to an underlying radiculopathy. For all other patients SDN is the treatment of choice. Following MTrP deactivation, correction of any postural disorder likely to cause MTrP reactivation is essential, as is the need to teach the patient how to carry out appropriate muscle stretching exercises. It is also important that the practitioner excludes certain biochemical disorders. PMID- 11926601 TI - Standards for reporting interventions in controlled trials of acupuncture: The STRICTA recommendations. STandards for Reporting Interventions in Controlled Trails of Acupuncture. AB - Acupuncture treatment and control group interventions in parallel-group randomised trials of acupuncture are not always precisely reported. In an attempt to improve standards, an international group of experienced acupuncturists and researchers devised a set of recommendations, designating them STRICTA: STandards for Reporting Interventions in Controlled Trials of Acupuncture. In a further consensus-building round, the editors of several journals helped redraft the recommendations. These follow the Consolidated Standards for Reporting Trials (CONSORT) format, acting as an extension of the CONSORT guidelines for the specific requirements of acupuncture studies. Participating journal editors are publishing the STRICTA recommendations and requesting prospective authors to adhere to them when preparing reports for publication. Other journals are invited to adopt these recommendations. The intended outcome is that interventions in controlled trials of acupuncture will be more adequately reported, thereby facilitating an improvement in critical appraisal, analysis and replication of trials. PMID- 11926602 TI - Moxibustion in breech version--a descriptive review. AB - The management of breech presentation at term remains controversial. It appears logical that maternal and perinatal outcomes would be improved if breech presentation could be avoided. External cephalic version is considered a safe procedure if cases are selected appropriately and anaesthesia avoided. Moxibustion is a traditional Chinese method of treatment, which utilizes the heat generated by burning herbal preparations containing the plant Artemisia vulgaris to stimulate the acupuncture points. It is used for breech version with a reported success rate of 84.6% after 34 weeks gestation. Moxibustion technique is cheap, safe, simple, self-administered, non-invasive, painless and generally well tolerated. Although many studies give encouraging results regarding the use of moxibustion in inducing cephalic version of breech presentation, a definitive conclusion cannot be made as most involve small sample sizes and are not randomised. Moxibustion could be an extra option offered to women with breech presentation along with vaginal delivery, caesarean section and external cephalic version. This article discusses the possible role of moxibustion in correction of breech presentation in the hope that, some interest will be stimulated in what is a very interesting area for future research. PMID- 11926603 TI - An audit of 500 acupuncture patients in general practice. AB - This is an audit of 500 patients treated in a general practice setting. Acupuncture clinics are provided on two evenings each week and are funded by the practice. Short treatment times are favoured but this varies according to condition and response. A wide variety of conditions have been treated the commonest being neck pain, low back pain, shoulder problems, hayfever, knee osteoarthritis, and migraine. An overall improvement of 73% was obtained with 61% classified as 'significant improvement' or 'cure'. Amongst the adverse effects recorded, retained needles and forgotten patients have been the most concerning and have led to changes in practice following 'critical incident' meetings. PMID- 11926604 TI - An audit of the effectiveness of acupuncture on musculoskeletal pain in primary health care. AB - Little is known about the use of acupuncture in general practice. We performed a retrospective review of the use of acupuncture in relieving musculoskeletal pain, a condition that is commonly encountered in general practice. A sample of 116 patient records was reviewed, from which 92 patients (mean age 52 years, 64% female) met the inclusion criterion of musculoskeletal pain. Information obtained included age, sex, diagnosis, duration of the problem, length of treatment (weeks), number of treatments, duration of each treatment (minutes), number of needles used, level of benefit obtained from the treatment, and recurrence of pain. There were many different conditions encountered. We found an association between the general practitioner using fewer needles and patients experiencing greater pain relief. This could be a reflection of treating myofascial pain syndromes, which often appear to respond well to a single needle in the key trigger point. Overall, we found that sixty-nine percent of patients had a good or excellent response to acupuncture treatment. We recommend acupuncture as a treatment option for patients who do not respond to the usual therapies (non steroidal anti-inflammatory drugs) for musculoskeletal conditions. PMID- 11926605 TI - Acupuncture for carpal tunnel syndrome. AB - Acupuncture was used to treat a 51-year-old 'lollipop lady' (school crossing patrol officer), with severe carpal tunnel syndrome (CTS) affecting her dominant hand, and co-existing cervical spondylosis. I postulate that her symptoms were work related. She responded well to acupuncture, which provided good symptomatic treatment rather than cure and allowed her to continue working whilst she awaited surgical release. PMID- 11926609 TI - Assessment and management of acute poisoning by petroleum products. AB - Petroleum products are highly complex chemical mixtures consisting predominantly of hydrocarbons. Their composition varies with source and intended use of the product. Virtually all are blended products that come into contact with man in a wide range of circumstances. Their toxicity for man is generally low and the use of additives rarely affects the toxicity of the final product. Because products are blended to meet performance, and not chemical specifications, their composition varies significantly. Management of toxicity benefits from simplified guidelines that consider the product by its type. Management in most cases is symptomatic, but the doctor needs to be aware of the potential for development of sequelae such as aspiration pneumonia and central nervous system (CNS) depression. Local and systemic effects of exposure to hydrocarbons are reviewed, as are immediate assessment and recommended management of acute exposure to petroleum products. Because of the large scope of this subject, this paper limits itself to acute toxicity of petroleum products encountered inthe public domain. It does not address topics such as chronic toxicity, solvent abuse, petrochemicals, or pesticides. PMID- 11926610 TI - Defining a tolerable concentration of methanol in alcoholic drinks. AB - Methanol, a potent toxicant in humans, occurs naturally at a low level in most alcoholic beverages without causing harm. However, illicit drinks made from "industrial methylated spirits" [5% (v/v) methanol:95% (v/v) ethanol] can cause severe and even fatal illness. Since documentation of a no-adverse-effect level for methanol is nonexistent in the literature a key question, from the public health perspective, is what is the maximum concentration of methanol in an alcoholic drink that an adult human could consume without risking toxicity due to its methanol content? Published information about methanol-intoxicated patients is reviewed and combined with findings in studies in volunteers given small doses of methanol, as well as occupational exposure limits (OELs), to indicate a tolerable ("safe") daily dose of methanol in an adult as 2 g and a toxic dose as 8 g. The simultaneous ingestion of ethanol has no appreciable effect on the proposed "safe" and "toxic" doses when considering exposure over several hours. Thus, assuming that an adult consumes 4 x 25-ml standard measures of a drink containing 40% alcohol by volume over a period of 2 h, the maximum tolerable concentration (MTC) of methanol in such a drink would be 2% (v/v) by volume. However, this value only allows a safety factor of 4 to cover variation in the volume consumed and for the effects of malnutrition (i.e., folate deficiency), ill health and other personal factors (i.e., ethnicity). In contrast, the current EU general limit for naturally occurring methanol of 10 g methanol/l ethanol [which equates to 0.4% (v/v) methanol at 40% alcohol] provides a greater margin of safety. PMID- 11926611 TI - Combined effects of acetaminophen, isopropylantipyrine and caffeine on pregnant and nonpregnant liver. AB - The common effects of acetaminophen (APA), isopropylantipyrine (IPA) and caffeine on liver were examined in rats. The preparations were given in Tween-80 solution once daily, in a constant proportion of 5:3:1, during days 8 to14 of gestation (S1, S2, S3 groups) or between days 8 and 14 of the experiment in nonpregnant female Wistar rats (S1-NP, S2-NP, S3-NP groups). The administration was in three different doses: doses S1, S1-NP-3.5 mg/kg APA, 2.14 mg/ kg IPA, 0.7 mg/kg caffeine; doses S2, S2-NP were 10 times higher; and doses S3, S3-NP 100 times higher than doses S1, S1-NP There were two control groups (T, T-NP) with Tween 80. At day 21 of gestation/experiment blood was taken for determination of activity of alanine (ALA) and aspartate (AST) aminotransferase, lactate (LDH) and glutamate (GLDH) dehydrogenase, levels of bilirubin (BIL), urea (URE), total protein (TP) and thymol turbidity test (TTT). The liver sections were examined by light microscopy with four stains: hematoxylin and eosin (H+E), silver Gomori, van Gieson and periodic acid-Schiff (PAS). There was a statistically significant (P<0.05) increase in the GLDH (S3-NP, S2, S3 groups) and AST, LDH (S3 group) activity, a decrease in URE (S2, S3 groups) and decrease in TP (S3-NP, S2, S3 groups) compared to the corresponding control group. Significant differences were noted in activity of AST, GLDH, and levels of the URE and TP between pregnant and nonpregnant females. The treatment resulted in minimal reactive and degenerative changes in light microscopic pattern of liver. PMID- 11926612 TI - Effects of cadmium on gap junctional intercellular communication in WB-F344 rat liver epithelial cells. AB - Cadmium has been associated with a number of tumors but its role in tumor promotion has not been elucidated clearly or the results obtained from various studies have been conflicting. This study was designed to investigate the effects of cadmium on the gap junctional intercellular communication (GJIC), number of gap junctions per cell, and cell proliferation in WB-F344 rat liver epithelial cells from the viewpoint of tumor promotion. GJIC was monitored by counting the cells stained with Lucifer yellow CH dye, using the scrape-loading and dye transfer method. The numbers of gap junctions per cell were visually quantitated after an indirect immunostaining for gap junction protein using an antibody to connexin 43. Cell proliferation was assayed by direct counting of the living cells using the trypan blue dye exclusion method. In the time course study, cells treated with 200 microM CdCl2 showed rapid and nearly complete inhibition of GJIC (approximately 14% of the control) and a decrease in the number of gap junctions per cell (approximately 21% of the control) at 30 min, and the decrease continued up to 4 h without any changes in the cell viability. Treatment with CdCl2 (7.4 200 microM) for 4 h resulted in the decrease of GJIC and gap junction numbers per cell in a dose-response pattern without changes in the cell viability. In the long-term (14 days) exposure studies at doses of 0.01-7.4 microM CdCl2, an increase in cell proliferation was observed at low doses of 0.03-2.5 microM CdCl2, with GJIC also decreasing. These data demonstrate that cadmium inhibits GJIC, reduces the number of gap junctions per cell, and induces cell proliferation while decreasing the function of the gap junction. PMID- 11926613 TI - Trichloroethylene induced testicular toxicity in rats exposed by inhalation. AB - Trichloroethylene (TCE) is an organic solvent used in dry cleaning, metal degreasing, thinner for paints and varnishes, anesthetic agent, and so forth. Human beings are appreciably exposed to TCE vapours by inhalation route. The present study has been undertaken to investigate whether TCE inhalation may also bring about testicular toxic effects. Our results indicate that inhalation of TCE by male rats for 12 and 24 weeks brings about significant reduction in absolute testicular weight, and alters marker testicular enzymes activity associated with spermatogenesis and germ cell maturation, along with marked histopathological changes showing depletion of germs cells and spermatogenic arrest. PMID- 11926614 TI - Positive responses to imipramine in the popliteal lymph node assay are due to primary irritation. AB - The popliteal lymph node (PLN) assay has long been proposed as a tool to detect immunotoxicants with the potential to induce systemic autoimmunity. A major problem hampering the further validation of this assay is the need to rule out irritants that cause false-positive PLN responses. The anti-depressant, imipramine, has not been reported to induce systemic autoimmune reactions in treated patients, but has been repeatedly found positive in the PLN assay, suggesting that this is a false-positive response. To test this hypothesis, the effects of imipramine were compared to those of 50% ethanol in C57B1/6 mice. Footpad edema was evidenced in the few days after injection of both ethanol and imipramine. T-cell depletion using monoclonal antibodies against either CD4+ or CD8+ T-lymphocytes prior to the PLN assay did not influence the responses to either ethanol or imipramine. Cytokine (TNFalpha, IL-1alpha, IL-1beta, IL-2R, IL 6, IL-12 and IFN-gamma) fingerprinting of the PLNs after injection of ethanol and imipramine evidenced the same pattern of responses. These results indicate a closely similar pattern of responses following the footpad injection of either imipramine or ethanol. The conclusion can be drawn that imipramine induces positive responses in the PLN assay via primary (nonspecific) irritation. PMID- 11926615 TI - Neutropenia induced by paraquat poisoning. AB - Paraquat is a synthetic, nonselective, contact herbicide, which causes injury to tissues following contact. When ingested in sufficient amounts, paraquat has life threatening effects on the gastrointestinal tract, lungs, kidney, liver, heart and other organs. This is a case report of a 15-year-old girl from an agricultural area who was admitted to hospital for treatment. She presented with discomfort, nausea and vomiting. She had drunk a small wine glass full of paraquat. After 18 days in the hospital, the girl developed neutropenia. This is the first reported case of paraquat poisoning-induced neutropenia. PMID- 11926616 TI - Replacing myth and prejudice with scientific facts about complementary and alternative medicine. PMID- 11926617 TI - Mother Nature strikes back: global warming, homeostasis, and implications for allergy. PMID- 11926618 TI - Innate immunity and its role against infections. AB - LEARNING OBJECTIVES: This article reviews current concepts of the innate immune system that offers protection against infections. It offers an overview for the readers to understand how innate immunity, consisting of different receptors, cells, and mediators recognizes pathogens and exerts protective function against pathogens. DATA SOURCES AND STUDY SELECTION: MEDLINE-search articles including original research papers, review articles, textbooks, and references identified from bibliographies of relevant articles. RESULTS AND CONCLUSIONS: The innate immune system is nonspecific immunity present since birth not requiring repeated exposure to pathogens. It is capable of differentiation between self and nonself. Because of its nonspecificity, it has a broad spectrum of resistance to infection. Further, it is thought to play an important role in the control of adaptive immunity by regulating co-stimulatory molecules and effector cytokines. Innate immunity includes pattern recognition molecules/receptors, antimicrobial peptides, the complement system, inflammatory mediators, and cytokines produced by immune cells. Pattern recognition molecules/receptors recognize pathogen associated molecular patterns that are essential for microorganisms' survival and pathogenicity. Although innate immunity has recently gained increasing importance, further studies are necessary for a better understanding of its role. PMID- 11926619 TI - Seven-year-old girl with joint stiffness, abnormal gait, and eosinophilia. PMID- 11926620 TI - Evaluation of the clinical efficacy and safety of grapeseed extract in the treatment of fall seasonal allergic rhinitis: a pilot study. AB - BACKGROUND: Herbal products are widely used by consumers as alternatives to prescription drugs in treating symptoms of allergic rhinitis. However, there have been few placebo-controlled clinical trials that have examined the efficacy or safety of these products. Although grapeseed extract (GSE) is an herbal that is marketed for treating allergic rhinitis, its efficacy is unproven. OBJECTIVE: The aim of this study was to compare the efficacy and safety of GSE with placebo in the treatment of seasonal allergic rhinitis (SAR). METHODS: This was a randomized, double-blind, placebo-controlled study of GSE 100 mg, twice daily, versus placebo. Patients with SAR and skin prick test sensitivity to ragweed were randomized to 8 weeks of active treatment or placebo which was begun before the ragweed pollen season. Outcomes included: daily symptom diary scores recorded by patients, rhinitis quality of life scores at baseline and after 4 and 8 weeks of treatment; and requirements for rescue chlorpheniramine. Safety was monitored with routine laboratory studies. RESULTS: Over 8 weeks of treatment, no significant differences were observed between active and placebo groups in rhinitis quality of life assessments, symptom diary scores, or requirements for rescue antihistamine. No significant laboratory abnormalities were detected. CONCLUSIONS: This study did not show trends supporting the efficacy of GSE in the treatment of SAR. PMID- 11926621 TI - Production of allergenic pollen by ragweed (Ambrosia artemisiifolia L.) is increased in CO2-enriched atmospheres. AB - BACKGROUND: The potential effects of global climate change on allergenic pollen production are still poorly understood. OBJECTIVE: To study the direct impact of rising atmospheric CO2 concentrations on ragweed (Ambrosia artemisiifolia L.) pollen production and growth. METHODS: In environmentally controlled greenhouses, stands of ragweed plants were grown from seed through flowering stages at both ambient and twice-ambient CO2 levels (350 vs 700 microL L(-1)). Outcome measures included stand-level total pollen production and end-of-season measures of plant mass, height, and seed production. RESULTS: A doubling of the atmospheric CO2 concentration stimulated ragweed-pollen production by 61% (P = 0.005). CONCLUSIONS: These results suggest that there may be significant increases in exposure to allergenic pollen under the present scenarios of global warming. Further studies may enable public health groups to more accurately evaluate the future risks of hay fever and respiratory diseases (eg, asthma) exacerbated by allergenic pollen, and to develop strategies to mitigate them. PMID- 11926622 TI - Symptom monitoring in childhood asthma: a randomized clinical trial comparing peak expiratory flow rate with symptom monitoring. AB - BACKGROUND: Accurate symptom evaluation is a critical component of asthma management. Limited data are available about the accuracy of symptom evaluation by children with asthma and their parents, or the impact of various symptom monitoring strategies on asthma morbidity outcomes. OBJECTIVE: The purpose of this randomized clinical trial was to evaluate the effect of three different intensities of symptom monitoring on asthma morbidity outcomes. METHODS: One hundred sixty-eight children (ages 6 to 19) of diverse racial, geographic, and socioeconomic backgrounds were randomized to 1 of 3 treatment groups (subjective symptom evaluation, symptom-time peak expiratory flow rate (PEFR) monitoring, daily PEFR monitoring) in this longitudinal, clinical trial. Outcome measures included a summary asthma severity score, forced expiratory volume in 1 second, symptom days, and health care utilization. RESULTS: Children who used PEFR meters (PFMs) when symptomatic had a lower asthma severity score, fewer symptom days, and less health care utilization than children in the other two treatment groups. Minority and poor children had the greatest amount of improvement using PFMs when symptomatic. Results were much less striking in white families. Thirty percent of families in the PFM treatment groups discontinued use entirely by 1 year postexit, whereas the majority of families who continued use (94%) used them only when symptomatic to inform symptom interpretation and management decisions. CONCLUSIONS: Not every child with asthma needs a PFM. Children and families facing extra challenges as a result of illness severity, sociodemographic, or health care system characteristics clearly benefited most from PFM use. PMID- 11926623 TI - A cross-sectional analysis of work-related outcomes in adults with asthma. AB - BACKGROUND: Asthma is a disease with physical symptoms that can impair a person's functioning to the point of interfering with school, work, and social activities. OBJECTIVE: Describe how disease (asthma) and patient characteristics affect perceived work performance and absenteeism. METHODS: Using a cross-sectional study design, 369 adults with asthma from a managed care organization responded to a mailed questionnaire which included a perceived work performance scale (WPS) and an item assessing absenteeism. Analysis consisted of psychometric testing of the scale (internal consistency and construct validity); trend observation of influence of perceived and symptom-derived severity on WPS and absenteeism distribution; and regression analysis to examine the relationship between patient/disease characteristics and the work-related outcomes. RESULTS: The mean WPS score was 88.0 +/- 16.2 (of a possible 100), with a Cronbach's alpha of 0.79. Most respondents (84.7%) did not miss any work in the previous 4 weeks. WPS scores declined and the percentage of respondent absenteeism increased as perceived and symptom-derived severity worsened. The regression model for WPS produced an adjusted R2 of 0.32 and included the number of other illnesses, health beliefs, race, income, and perceived and symptom-derived asthma severity. The regression model for absenteeism included number of other illnesses, race, health beliefs, and symptom-derived severity. Perceived work performance and absenteeism are outcomes measures that are distinctive and complementary. CONCLUSIONS: Disease severity, race, income, and health beliefs also contribute and should be considered in health services research related to asthma. PMID- 11926624 TI - Efficacy of dry-cleaning in removing Fel d 1 allergen from wool fabric exposed to cats. AB - BACKGROUND: The main cat allergen (Fel d 1) is ubiquitous, having been found even in indoor environments and public places where a cat has never been kept. Clothes of cat owners constitute a carrier for the distribution of Fel d 1 allergen in these environments. Schools, for example, may be a site of indirect exposure to cat allergens. OBJECTIVE: Our goal was to investigate the efficacy of commercial dry-cleaning in removing cat allergens from wool fabrics that had been exposed to cats to evaluate a possible preventive procedure. METHODS: Twenty-six identical wool "squares" (80 x 100 cm) were put in cat baskets for 1 week. In our laboratory, the squares were cut in half (40 x 50 cm), and one half was subjected to high-volume sampling for 5 minutes in a cat-free room. The other half was subjected to commercial dry-cleaning and then the high-volume sampling. Five wool squares not exposed to cats served as controls. Dust was collected from the wool squares with a high-volume air sampler. Particulate material was harvested onto glass fiber filters (AP 20 Millipore, Milan, Italy) with 25-mm diameter and 2 microm pore size. Each dust sample was assayed by affinity-purified monoclonal antibody against purified Fel d 1. The results were expressed as micrograms per filter. Statistical analysis was done by using the paired t test. RESULTS: Before dry-cleaning, Fel d 1 allergen was detected on all cat-exposed wool squares. No appreciable cat allergen was detected on control materials. After commercial dry cleaning, the amounts of Fel d 1 extracted from cat-exposed squares were significantly reduced (t = 14.63; P < 0.001) but not abolished. Three of the five control squares were contaminated by Fel d 1. CONCLUSIONS: Commercial dry cleaning effectively removes large amounts of cat allergen from wool materials exposed to cats but does not completely abolish this protein. Further, low Fel d 1 contamination may occur during this procedure. PMID- 11926625 TI - Effects of budesonide inhalation suspension on hypothalamic-pituitary-adrenal axis function in infants and young children with persistent asthma. AB - BACKGROUND: The initial 12-week, double-blind phases of three studies demonstrated that budesonide inhalation suspension (BIS) is effective and well tolerated in infants and young children (6 months to 8 years of age) with persistent asthma. OBJECTIVE: Open-label, 52-week extensions to these studies were conducted to evaluate long-term safety of BIS, including effects of treatment with the lowest effective dose of BIS on hypothalamic-pituitary-adrenal (HPA)-axis function, as compared with conventional asthma therapy (CAT). Complete results of the earlier phases of the studies and of long-term safety are reported elsewhere; only results pertaining to HPA-axis function are summarized here. METHODS: Patients eligible for the open-label phases of the three trials were randomized to treatment with nebulized BIS (n = 447) or CAT (n = 223). CAT included short-acting oral or inhaled beta2-agonists, methylxanthines, or cromolyn sodium; in two of the studies, CAT could have included other inhaled corticosteroids. HPA-axis function, which had been evaluated during the 12-week double-blind studies, was again evaluated at the beginning and end of the 52-week study period using basal plasma cortisol concentrations and response to stimulation with a 250-microg dose of adrenocorticotropic hormone. RESULTS: There was no evidence of altered HPA-axis function attributable to BIS treatment. No clinically or statistically significant differences in basal or adrenocorticotropic hormone-stimulated plasma cortisol concentrations were observed between BIS and CAT in either the 12-week, double-blind or 52-week, open label phases of the three studies. CONCLUSIONS: The results indicate that treatment with BIS does not result in clinically significant suppression of HPA axis function in infants and young children. PMID- 11926626 TI - Idiopathic anaphylaxis: a descriptive study of 81 patients in Spain. AB - BACKGROUND: Few European groups have published data on series of patients with idiopathic anaphylaxis. OBJECTIVE: Our aim was to report on our experience with this entity in Spain. METHODS: We conducted a two-part investigation. The first part was a descriptive cross-sectional study of 81 patients with idiopathic anaphylaxis, diagnosed between January 1990 and December 1995 in the allergy unit of the General Hospital of Albacete in Albacete, Spain. The second prong was a prospective, longitudinal study that evaluated the activity of the condition. RESULTS: Female patients constituted 68% of the study group with idiopathic anaphylaxis; the mean age of the 81 patients was 30.0 +/- 17.3 years. The number of episodes of idiopathic anaphylaxis during the year of greatest frequency varied widely (median, 2; range, 1 to 130). The duration also varied considerably, with a range of 0.03 to 300 months (median, 2 years). Most patients entered the study in remission, and 93% of these patients remained in remission throughout the followup period. Only 9% of the patients had vascular involvement. Our series of patients with idiopathic anaphylaxis had a notable prevalence of atopic diseases (48%), food allergy (20%), and episodes of anaphylaxis with an identifiable cause (15%). In addition, a high frequency (58%) of idiopathic urticaria was noted; the urticaria was acute in 62% of the affected patients. CONCLUSIONS: Our series of patients with idiopathic anaphylaxis is similar to previously published series. PMID- 11926628 TI - Multicolored simplified asthma guideline reminder (MSAGR) for better adherence to national/global asthma guidelines. AB - BACKGROUND: Clinicians in general have not widely and consistently used asthma guidelines in their practices around the world. This study identifies reasons for the poor adherence to asthma guidelines by primary care physicians (PCPs), and simultaneously introduces multicolored simplified asthma guideline reminder (MSAGR) as a practical tool to enhance adherence to asthma guidelines. METHODS: Sixty-nine PCPs were given a simple, one-page, fill-in-the-blank questionnaire on the classification of asthma severity as defined in National Asthma Education and Prevention Program guidelines, using patients' symptoms, peak expiratory flow rate (PEFR)/forced expiratory volume in 1 second (FEV1) value, PEFR variability, and step therapy based on asthma severity. Also, they were given a questionnaire on barriers to using asthma guidelines and MSAGR for evaluation. In one targeted community, free copies of MSAGR were made available to PCPs, and data on emergency room visits and hospitalization of asthmatic patients were analyzed. RESULTS: Of the PCPs, 16% correctly classified mild, intermittent asthma, 13% mild, persistent asthma, 8% moderate, persistent asthma, and 8% severe, persistent asthma based on the combined patient's symptoms, PEFR or FEV1 value and PEFR variability as defined in National Asthma Education and Prevention Program guidelines. One hundred percent of the PCPs chose inhaled beta2-agonists as quick relief medication. Fifty percent of the PCPs chose inhaled steroids, leukotriene antagonists, oral theophylline, and long acting beta-agonists in various combinations for different severity of asthma. Eighty percent of the physicians failed to select the appropriate dosages of inhaled steroids for different severities of asthma. Ninety-five percent of PCPs reported that MSAGR made using the guidelines easier for them. In the targeted community, asthma related emergency room visits decreased 22.5% and hospitalizations by 26.9%. CONCLUSIONS: This is the first study that identified the reasons for poor adherence to asthma guidelines by PCPs, and introduced MSAGR as a practical "low tech" tool to promote better adherence to asthma guidelines. MSAGR presents patient-specific recommendations, based on asthma guidelines in a user-friendly format that can save the physician time in real-world primary care settings, where such information is often needed instantly. The overwhelming majority of PCPs strongly agreed that MSAGR helped them recall the classification of asthma severity in a timely manner, to inquire about various triggers, and to use step therapy accurately and confidently. In one targeted community, MSAGR helped clinicians in primary care settings to achieve better asthma outcomes and to reduce both emergency room visits and hospitalizations. PMID- 11926627 TI - Prevalence of serious eosinophilia and incidence of Churg-Strauss syndrome in a cohort of asthma patients. AB - BACKGROUND: Some leukotriene receptor antagonists, such as zafirlukast and montelukast, have been associated with systemic eosinophilia, with interest focused on Churg-Strauss syndrome (CSS). OBJECTIVE: To calculate the background incidence rate of CSS and prevalence of eosinophilia among people with asthma who have not used leukotriene receptor antagonists. METHODS: We conducted a cohort study in the setting of three geographically diverse UnitedHealthcare health plans. We identified 36,230 people who received a diagnosis of asthma during the period October 1994 through September 1997. We identified 241 potential cases from the claims data. Using a standardized hospital record abstract form, nurses abstracted relevant clinical data from the hospital charts of potential cases. We applied several a priori case definitions to the abstracted clinical data and computed incidence rates of CSS among patients with asthma. We additionally used these data to compute the prevalence of serious eosinophilia. RESULTS: Incidence rates of definite CSS among asthma patients ranged from zero (90% confidence interval 0.0 to 23.0) to 67 (90% confidence interval 22.5 to 160.6) cases per 1,000,000 person-years, depending on the definition used. All patients who met the criteria for CSS expressed symptoms consistent with mild asthma. CONCLUSIONS: This report is the first direct measurement of the incidence rate of CSS among asthma patients. We believe that the prevalence and incidence information that we report is a useful description of population rates in the United States for these conditions. PMID- 11926629 TI - Tolerability of rofecoxib in patients with cutaneous adverse reactions to nonsteroidal anti-inflammatory drugs. AB - BACKGROUND: Adverse reactions to nonsteroidal anti-inflammatory drugs (NSAIDs) are very common problems in clinical practice. Rofecoxib is a novel NSAID which selectively inhibits prostaglandin endoperoxide H synthase-2 and has no effect on prostaglandin endoperoxide H synthase-1. OBJECTIVE: To evaluate the tolerability of rofecoxib in NSAID-sensitive patients with cutaneous adverse reactions. PATIENTS AND METHODS: One hundred thirty-nine patients with NSAID-induced adverse reactions [60 urticaria alone (43.1%), 34 angioedema (24.5%), 34 angioedema plus urticaria (24.5%), 5 urticaria plus difficulty in breathing (3.6%), 4 maculopapular rush (2.9%), and 2 Stevens-Johnson syndrome (1.4%)] were submitted to a single-blind, placebo-controlled peroral challenge with increasing doses of rofecoxib. RESULTS: One hundred thirty-eight of 139 (99.3%) patients tolerated rofecoxib without adverse reactions; only one (0.7%) experienced weak urticaria localized at arms. CONCLUSIONS: Rofecoxib is a well tolerated drug in patients with NSAID cutaneous adverse reactions and it may represent a valid alternative drug in NSAID-sensitive patients. PMID- 11926630 TI - Trichosporon asahii-induced asthma in a family with Japanese summer-type hypersensitivity pneumonitis. AB - BACKGROUND: Trichosporon species frequently induce summer-type hypersensitivity pneumonitis (SHP), which is the most prevalent type of hypersensitivity pneumonitis (HP) in Japan, but have not been reported to induce asthma. OBJECTIVE: Evaluation of a case of asthma induced by Trichosporon asahii. METHODS AND RESULTS: This report describes a 46-year-old Japanese man who developed asthma induced by T. asahii, following symptoms of HP attributable to the same pathogen, in a case of familial occurrence of SHP. This patient lacked typical findings of HP in his radiograph but had an elevated level of eosinophils in his bronchoalveolar lavage fluid. Open lung biopsy, however, revealed typical pathologic findings of HP when he was free of asthmatic symptoms. His serum was also positive for anti-T asahii antibody, as are the sera of SHP patients. Nevertheless, provocation tests, including returning home and inhalations of T. asahii antigen, reproduced asthmatic features such as airway hyperresponsiveness and reversible bronchoobstruction, but not the features of HP. A skin test with the same antigen also evoked an immediate allergic reaction. An IgE mechanism was suspected but could not be proven by radioallergosorbent test. The patient's son and daughter displayed typical features of SHP, associated with compatible results in their radiographs, bronchoalveolar lavage fluid analysis, serologic and pathologic examinations, and provocation and skin tests. CONCLUSIONS: To our knowledge, this is the first case of extrinsic asthma, and of coexistent asthma and HP, induced by T. asahii. The patient initially displayed symptoms typical of SHP, which were subsequently replaced by more typical asthmatic symptoms. PMID- 11926631 TI - Primary leptomeningeal lymphoma in a patient with concomitant CD4+ lymphocytopenia. AB - BACKGROUND: Idiopathic CD4+ lymphocytopenia (ICL) is a rare disorder in which patients have mild and/or severe opportunistic infections or maybe without symptoms. The etiology is currently unknown. Diagnosis is made by excluding retroviral infections (human immunodeficiency virus-1 or -2, human T cell lymphotropic virus-1 or -2) or other known causes of immunosuppression. OBJECTIVE: To provide a case report of a patient with possible ICL who presented with a rare form of primary non-Hodgkin lymphoma (NHL) of the central nervous system (CNS). Review of the literature has identified only five other patients with NHL and ICL; however, none of these had a CNS lymphoma. RESULTS: We describe a patient with possible ICL, and address links between lymphopenia and lymphoproliferative disorders. CONCLUSIONS: Although not uncommon for patients infected with human immunodeficiency virus to develop CNS NHL, this is the first case of a possible ICL patient with such a lymphoma. This case revisits an important relationship between lymphopenia and lymphoproliferative disorders. PMID- 11926632 TI - Bermuda grass. PMID- 11926634 TI - Short-term neonatal outcomes of growth restricted infants by their mode of delivery. PMID- 11926633 TI - Eugenics or empowered choice? Community issues arising from prenatal testing. AB - The prevention of inherited disabilities is viewed in two contrasting ways- either as enhancing reproductive choice and improving population health, or as discriminating against disabled community members. We argue that modern clinical genetics, including preimplantation genetic diagnosis (PGD), reflects a persistent and defensible desire by the community to prevent disability, rather than as increasing discrimination or threatening to produce a 'new eugenic' society Screening should be presented as a distinct issue for decision-making about the prevention or acceptance of disability, rather than as a routinely accepted component of antenatal care. The community must improve its understanding of the experiences of those who manage disability, and continue to debate the issues of discrimination, selective genetic prevention and enhancement, reproductive freedom, and eugenics. PMID- 11926635 TI - Australian Society of Ultrasound in Medicine (ASUM) Policy Statement on normal ultrasonic fetal measurements. PMID- 11926637 TI - Audit of maternal and fetal outcomes in women treated for glucose intolerance during pregnancy. AB - OBJECTIVE: To determine whether one should aim for glycaemia that is statistically 'normal' or for levels of glycaemia low enough to prevent macrosomia (if such a threshold exists) when glucose intolerance is detected during pregnancy. DESIGN: An audit of pregnancy outcomes in women with impaired glucose tolerance in pregnancy as compared to a local age-matched reference group with normal glucose tolerance. RESULTS: Our study suggests that for most patients, more intensive therapy would not have been justified. Maternal smoking appeared to convey some 'advantages' in terms of neonatal outcomes, with reduction in large-for-gestational-age (LGA) infants and jaundice in babies of impaired glucose tolerance (IGT) mothers. CONCLUSIONS: These observations demonstrate the importance of considering risk factors other than GTT results in analysing pregnancy outcomes, while emphasising that 'normalisation' of fetal size should not be our only therapeutic endpoint. Our detailed outcome review allows us to reassure patients with GDM that with current treatment protocols, they should have every expectation of a positive pregnancy outcome. PMID- 11926636 TI - Legal abortion in South Australia: a review of the first 30 years. AB - OBJECTIVES: To review the first 30 years' experience of legal abortion in South Australia and its demographic implications. DATA AND METHODS: Information was obtained from official abortion statistics and demographic publications of the Australian Bureau of Statistics. Standard demographic and statistical techniques of analysis were used. RESULTS: After an initial rise during the 1970s, abortion rates remained fairly constant for the next decade but have increased since 1990. The Pregnancy Advisory Centre opened in 1992, to reduce waiting times and to cater for late abortions. This resulted in an increase in abortions earlier in pregnancy and also an increase in late abortions. With the adoption of vacuum aspiration techniques and improved services, abortion is now a day-only procedure, performed by specially trained doctors. Morbidity and mortality have been greatly reduced. Concurrent sterilisation has also declined. The increase in abortion has affected all age groups, but particularly women under 30, consistent with the national trend towards the postponement of births. After an initial rapid decline, the total pregnancy rate has risen slightly since 1990, showing changes in patterns of contraceptive use. However, this is not reflected in an increase in the total confinement rate. CONCLUSION: Overall, contraception has had a greater effect than abortion in reducing births in South Australia. The abortion rate is still lower than in the rest of Australia as calculated from Medicare data, even though this is an underestimate because it includes only fee paying patients. There remains a need for continuing emphasis on better contraceptive use, including emergency contraception. PMID- 11926638 TI - Gestational diabetes versus obesity as risk factors for pregnancy-induced hypertensive disorders and fetal macrosomia. AB - OBJECTIVE: To assess whether obesity and/or gestational diabetes mellitus (GDM) are independent risk factors for fetal macrosomia and/or pregnancy-induced hypertensive disorders. DESIGN: Retrospective computerised database review. SETTING: Lyell McEwin Health Service, South Australia. SAMPLE: All nulliparous women delivering singleton babies in the years 1999 and 2000 MAIN OUTCOME MEASURES: Birth weights, the occurrence of fetal macrosomia, gestational hypertension and preeclampsia were compared between 258 normoglycaemic women (control group), 76 women with only an abnormal glucose challenge test, and 51 women with GDM. RESULTS: Only GDM with fasting hyperglycaemia is a risk factor for macrosomia (risk ratio: 3.3 95% confidence limits 1.229-8.736). Smoking is associated with a decrease in the incidence of pregnancy-induced hypertensive disorders. CONCLUSIONS: Our data strongly suggest that GDM and obesity are not independent risk factors for pregnancy-induced hypertensive disorders. PMID- 11926639 TI - Caesarean delivery and outcome in very low birthweight infants. AB - OBJECTIVE: To determine the relationship between mode of delivery, intraventricular haemorrhage (IVH), and mortality in very low birthweight (VLBW) infants. STUDY DESIGN: A historical cohort study of infants admitted to a single level III neonatal intensive care unit during a five-year period. Infants < 1500 g born by caesarean delivery (n = 400) were compared to those born by vaginal delivery (n = 305). RESULTS: After controlling for potential confounding variables including: gestational age, fetal presentation, and multiple birth, caesarean delivery was not associated with a decreased odds of IVH (odds ratio 1.2, 95% CI 0.7-2.0), severe IVH (1.9, 0.9-4.0), or mortality (1.2, 0.6-2.4). CONCLUSIONS: In our population of very low birthweight infants, caesarean delivery is not associated with a decreased risk for mortality or intraventricular haemorrhage. PMID- 11926640 TI - Risk factors for preterm delivery in patients demonstrating sonographic evidence of premature dilation of the internal os, prolapse of the membranes in the endocervical canal and shortening of the distal cervical segment by second trimester ultrasound. AB - OBJECTIVE: To investigate the outcomes and risk factors associated with the second trimester sonographic findings of premature dilation of the internal cervical os, prolapse of the membranes into the endocervical canal and shortening of the distal segment. STUDY DESIGN: Retrospective chart review to identify patients who met the following criteria: (i) gestational age between 16 and 24 weeks; (ii) a vaginal probe ultrasound documenting dilatation of the internal os, prolapsed membranes into the endocervical canal but not extending beyond the external os, and a shortened distal cervix; and (iii) no evidence of increased uterine activity METHODS: Sonographic measurements obtained included: (i) the width of the internal os dilation; (ii) the depth of membranes prolapsed into endocervix (funneling); (iii) the distal cervical length; and (iv) the total cervical length. Patients were stratified into two groups according to gestational age at delivery: the preterm group (< 34 weeks) and the near-term group (> or = 34 weeks). RESULTS: Thirty-seven patients had complete records for analysis and their pregnancies resulted in 47 live births. Eighteen patients were stratified to the near term group and 19 were stratified to the preterm group. There were no stillborn infants and 10 neonatal deaths. Ten patients received cerclage and seven patients delivered at < 34 weeks gestational age. Risk factors found to be significant for preterm delivery included a gestational age at diagnosis of < or = 20 weeks gestation (p = 0.03), dilatation of the internal os > 1.3 cm (p = 0.04), and a composite Benham score > or = 1.0 (p = 0.02). CONCLUSIONS: The sonographic findings of premature dilatation of the internal os, prolapse of the membranes into the endocervical canal and shortening of the distal cervix are associated with a high rate of delivery < 34 weeks (51%) and neonatal death (27%). PMID- 11926641 TI - Higher order multiple pregnancies in New South Wales 1990-1999. AB - OBJECTIVE: To examine higher order multiple pregnancy and birth rates in NSW, and to describe trends in the characteristics and management of these births. DESIGN: Cross-sectional analytic study. SETTING: New South Wales, Australia. POPULATION: Two hundred and ninety-one women who gave birth to triplets, quadruplets or quintuplets (880 babies) from 1 January 1990 to 31 December 1999. METHODS: Data were obtained from the NSW Midwives Data Collection and rates over time were calculated. MAIN OUTCOME MEASURES: Higher order multiple birth and pregnancy rates (> or = 20 weeks), place of birth, mode of delivery, fetal death rates and Apgar scores. RESULTS: There was no significant change in the number or rate of higher order multiple births in NSW during the 1990s with an average annual rate of 10.3/10,000 births or 3.5/10,000 pregnancies > or = 20 weeks. Among women with higher order multiple pregnancies, those aged > or = 35 years increased from 19% in 1990 to 47% in 1999. There was also a trend towards delivery in a perinatal centre from 56% to 70%, and vaginal birth from 18% to 28%. There were no significant changes in infant outcomes. CONCLUSIONS: The increases in higher order multiple pregnancies observed in Australia in the 1980s, and into the 1990s in other countries, have not persisted in NSW, suggesting that guidelines for limiting the number of embryos/oocytes transferred in assisted reproductive technologies have been widely adopted. PMID- 11926643 TI - The impact of intrapartum analgesia on labour and delivery outcomes in nulliparous women. AB - OBJECTIVE: To determine if nulliparous women intending to have epidural analgesia have a similar labour profile and delivery outcome to women who intend to have their labour managed using alternative forms of pain relief. MATERIALS AND METHODS: A prospective randomised controlled clinical trial conducted at a tertiary obstetric institution. Nulliparous women intending to deliver vaginally with a term singleton fetus were eligible for recruitment. RESULTS: 1159 women were recruited, of whom 992 were subsequently randomised to receive continuous midwifery support (CMS) or epidural analgesia (EPI) on presentation for delivery. The duration of labour was shorter in the CMS group compared with EPI (10.7 hours (inter quartile (IQ) 7.0,15.2) versus 11.4 hours (IQ 8.2,15.2), p = 0.039). The median duration of the first stage was 8.9 hours (IQ 6,12.5) versus 9.5 hours (IQ 7,12.7) (p = 0.069), and the median duration of the second stage was 1.33 hours (IQ 0.6,2.5) versus 1.48 hours (IQ 0.77,2.6) (p = 0.034). The requirement for oxytocin augmentation in spontaneous labour was 39.8% CMS versus 46.2% EPI (p = 0.129). There was no significant difference in the caesarean section rates. The need for any operative delivery was significantly lower in CMS (43.9% CMS versus 51.5% EPI, p = 0.019). CONCLUSIONS: Nulliparous women have a high usage of epidural analgesia, regardless of their prelabour intentions. In women who do not intend to use epidural analgesia, the temporal delay in insertion compared with those who use epidural analgesia as their primary analgesic modality is associated with a small but statistically significant reduction in overall labour duration and operative delivery rates. PMID- 11926642 TI - The loss rates for invasive prenatal testing in a specialised obstetric ultrasound practice. AB - OBJECTIVES: To establish the spontaneous miscarriage rate and compare it with the procedure related miscarriage rate for amniocentesis and chorionic villus sampling (CVS) by experienced operators. DESIGN: Retrospective audit over a two year period of all patients having a consultation for prenatal diagnosis before 12 weeks gestation. SETTING: A specialised obstetric and gynaecological ultrasound practice. POPULATION: A total of 2366 patients, mostly over 35 years of age. METHODS: Between 1 July 1995 and 30 June 1997, all patients having a prenatal consultation decided between amniocentesis, CVS or no invasive testing. The CVS was performed either transabdominally or transcervically, depending on the position of the uterus and placenta. MAIN OUTCOME MEASURES: Delivery, termination of pregnancy for chromosomal abnormality or miscarriage. RESULTS: Over the two-year period, 2366 patients had a prenatal consultation and outcome data were available for all but 53 patients. After consultation, 346 patients decided not to have any prenatal testing and 29 (8.4%) of these had a spontaneous miscarriage. Amniocentesis was requested by 839 patients; however 10 miscarried before the scheduled procedure. After the amniocentesis, there were 13 terminations for chromosomal abnormality and three miscarriages. CVS was requested by 1128 patients; however, 23 miscarried before the scheduled procedure. Transabdominal CVS was performed in 665 patients, transcervical in 416 and in 24 cases the documentation of the method used was unclear. Eleven patients miscarried after the transabdominal CVS (1.65%) compared with nine patients miscarrying after the transcervical CVS (2.16%), which was not statistically significant (p = 0.27). CONCLUSION: In the group studied, the spontaneous miscarriage rate from nine weeks gestation is very high (8.4%). The procedure related loss rate from amniocentesis was less than 1 in 280. Transabdominal CVS appears to have a lower fetal loss rate than transcervical CVS, but much larger numbers are needed to prove this. PMID- 11926645 TI - Strategies required to improve the management of chronic vulvar discomfort in Australia. AB - OBJECTIVE: To investigate factors required to improve the management of vulvar pain from a patient perspective. SAMPLE AND SETTING: Sixty patients in four different specialised vulvar clinics. METHODS: Women with chronic vulvar pain were asked to complete a questionnaire whilst waiting for specialist consultation by either a dermatologist, sexual health physican, psychologist or physiotherapist. RESULTS: The average length of time patients had vulvar pain was 6.03 years, with a mean of 5.83 practitioners seen. Twenty-seven per cent indicated that increased awareness of vulvodynia was required by gynaecologists and 74% suggested that increased general practitioner awareness was necessary to improve care. Accurate diagnosis, and understanding lifestyle implications were key elements of patient satisfaction, considered more important than any available treatment modality. CONCLUSIONS: Current services and skilled practitioners available for the management of chronic vulvar pain or vulvodynia in Australia do not reflect the estimated prevalence of this condition. From this study it was concluded that the three most important factors required to improve quality of care in order of importance were increased practitioner awareness, research and the setting up of more specialised multi-disciplinary clinics. PMID- 11926644 TI - High-grade vulval intraepithelial neoplasia (VIN 3): a retrospective analysis of patient characteristics, management, outcome and relationship to squamous cell carcinoma of the vulva 1989-1999. AB - OBJECTIVES: To determine patient and disease characteristics, treatment patterns and long-term outcomes, in order to help direct management of patients with VIN 3, to examine the risk of development of cancer following a diagnosis of VIN 3 and the risk of recurrent disease requiring multiple treatment episodes, and to review patients with vulval cancer, in order to establish the relative prevalence of VIN 3 related vulval cancers. DESIGN: Retrospective chart review of 65 consecutive patients diagnosed with VIN 3 between 1989 and 1999. POPULATION OR SAMPLE: All patients diagnosed with VIN 3 at Christchurch Womens' Hospital (1 January 1989 to 31 December 1999) and Dunedin Public Hospital (1 January 1990 to 31 December 1999). MAIN OUTCOME MEASURES: Age at diagnosis, symptoms, medical history disease characteristics, treatment, outcome and follow-up. RESULTS: The median age at diagnosis was 38 years. Smokers were younger than non-smokers. Two thirds had associated dysplasia of the lower genital tract, 43% had high-grade lesions. Seventy-nine per cent were symptomatic for a median of 9.5 months. All had macroscopically visible disease. Colposcopy and histology diagnoses correlated in 72% of cases. Treatment by local excision was undertaken in 84% of cases of which 65% had involved margins. Fifty-one per cent required further treatment, risk factors were positive margins and multifocal disease. Three developed micro-invasive cancer. CONCLUSIONS: Treatment by local excision is both diagnostic and therapeutic. Excision may reveal micro-invasive cancer, patients are usually symptomatic and VIN 3 has some potential to become invasive. Treated patients may develop microinvasive disease but frank invasion was not seen. The true rate of malignant progression in untreated patients remains unclear and radical surgery is rarely indicated. All patients require long-term follow up. PMID- 11926646 TI - In utero fetal muscle biopsy in the diagnosis of Duchenne muscular dystrophy. PMID- 11926647 TI - Placenta percreta: four cases in one month. PMID- 11926648 TI - A reassessment of options for the management of placenta praevia percreta. PMID- 11926649 TI - Pregnancy in a woman with Friedreich's ataxia complicated by pulmonary embolism. PMID- 11926650 TI - Fetomaternal haemorrhage--can we improve the outcome? PMID- 11926651 TI - A possible role for implantable naltrexone in the management of the high-risk pregnant heroin user. PMID- 11926652 TI - Second trimester fetal death caused by Citrobacter koseri (diversus) infection: a report of two cases. PMID- 11926653 TI - Methicillin-resistant Staphylococcus aureus chorioamnionitis: a rare cause of fetal death in our community. PMID- 11926654 TI - The integral theory of pelvic floor dysfunction. PMID- 11926655 TI - The integral theory of female urinary incontinence. PMID- 11926657 TI - Pharmacist-physician collaboration. PMID- 11926656 TI - Age, gender, and the workforce. PMID- 11926658 TI - A pharmacy success story: 104,644 pharmacy technicians certified nationwide. PMID- 11926659 TI - Tablet splitting: imperfect perhaps, but better than excessive dosing. PMID- 11926660 TI - Great moments in pharmacy: development of the Robert Thom series depicting pharmacy's history. PMID- 11926661 TI - Beyond the 4Ps: using relationship marketing to build value and demand for pharmacy services. PMID- 11926662 TI - Lack of medication dose uniformity in commonly split tablets. AB - OBJECTIVE: To divide 11 commonly split tablets and evaluate the resulting half tablets for content uniformity. DESIGN: Pre-post comparison. SETTING: Laboratory. INTERVENTIONS: A trained individual split tablets of 11 products using a single edged razor blade and 3 products by hand alone. MAIN OUTCOME MEASURES: The Uniformity of Dosage Units test published in the United States Pharmacopeia 24 (USP), which applies to whole tablets, was adapted liberally to assess the dose uniformity of the resulting split tablets. RESULTS: Of the 11 razor-split products, 8 failed the liberal adaptation of the USP uniformity test. No visible tablet features (e.g., scoring) predisposed a product's split tablets to pass or fail the uniformity test. All three hand-split tablets failed the uniformity test and yielded worse results than did razor-split tablets. CONCLUSION: The majority of the 11 drug products we tested, when assessed for their ability to be split into half-tablets of equal dose, failed a liberally interpreted USP uniformity test. The practice of dividing tablets to save costs or to improve a dosage regimen may not cause problems for patients using drugs with low toxicity and relatively flat dose-response relationships, but it is not recommended for patients using drugs with more substantial toxicity and steep dose-response efficacy curves. PMID- 11926663 TI - Weight variability of pharmacist-dispensed split tablets. AB - OBJECTIVE: To determine the level of weight uniformity of fragments from tablets split into halves and dispensed by pharmacists. DESIGN: Pre-post comparison. SETTING: Laboratory analysis of split tablets returned unused from four long-term care facilities. INTERVENTIONS: Twenty-two dispensed prescriptions containing 560 split tablet halves were collected, and fragment weights (FWs) were determined. Theoretical weights (TWs) of split tablets of the same medications were determined mathematically. MAIN OUTCOME MEASURES: Comparison of the mean FW with the mean TW of each dispensed prescription and determination of whether fragments met United States Pharmacopeia 24 (USP) criteria for weight variation of whole tablets. RESULTS: A significant difference (P < .05) between mean TW and mean FW was found in 2 (9.1%) of the 22 dispensed prescriptions. Overall, 30 (5.4%) of 560 fragments, contained in 10 (45.5%) of the 22 dispensed prescriptions, had weights that deviated by more than 15% from the mean FW. Of the 560 fragments, 32 (5.7%), contained in 10 (45.5%) of 22 dispensed prescriptions, had weights that deviated by more than 15% from mean TW. A total of 15 (68.2%) of the 22 dispensed prescriptions had standard deviations (SDs) that were more than 6% of the mean FW. None of the TWs displayed an SD greater than the USP limit of 6%. Only 7 (31.8%) of the 22 dispensed prescriptions met USP standards, with FWs not exceeding 15% of mean FW and SDs of no greater than 6%. CONCLUSION: Tablet splitting resulted in an unacceptably high incidence of weight variation. Standards should be developed to ensure uniformity of split tablets. PMID- 11926664 TI - Healthy skin for women: a review of common conditions and therapies. AB - OBJECTIVE: To review common dermatologic conditions that may be of particular concern to women throughout their lives, including acne, contact dermatitis, skin conditions of pregnancy, dry skin, and sun-related skin damage. DATA SOURCES: Articles identified through a search of MEDLINE and pharmacology and dermatology textbooks using the terms women, dermatology, skin, acne, skin cancer, sunscreens, pregnancy, melanoma, basal cell carcinoma, dermatitis, cosmetic dermatitis, dry skin (xerosis), actinic keratosis, oral contraceptives, and striae. Additional references were identified from the bibliographies of retrieved articles and book chapters. DATA SYNTHESIS: A number of skin conditions may be of particular concern to women throughout their lives. Acne, which may be particularly troublesome during the teenage years and young adulthood, and skin conditions that are common during pregnancy may be influenced by hormonal changes that occur at different life stages. Skin conditions that are influenced by the environment include contact dermatitis, dry skin, and sun-related skin damage, including actinic keratosis, carcinoma, and melanoma. Many of these conditions are amenable to nonpharmacologic therapy and over-the-counter treatments, whereas others require prescription pharmacotherapy. Pharmacists are in an ideal position to advise women about the appropriate management of these conditions and to identify lesions that require additional assessment. CONCLUSION: Pharmacists can make valuable contributions to patients' self-care of the skin by recommending products, educating patients about skin health, and counseling patients about prescription medications. Gaining a greater understanding of skin conditions that are of particular concern to women can prepare pharmacists to make these contributions. PMID- 11926665 TI - Natural products used for diabetes. AB - OBJECTIVE: To review the efficacy and safety of natural products commonly used for diabetes. DATA SOURCES: English and Spanish-language journals retrieved through a MEDLINE search of articles published between 1960 and December 2001 using these index terms: Opuntia, karela, gymnema, tecoma, alpha lipoic acid, thioctic acid, ginseng, panaxans, and diabetes. DATA SYNTHESIS: Natural products have long been used in traditional systems of medicine for diabetes. Products in common use include nopal (prickly pear cactus), fenu-greek, karela (bitter melon), gymnema, ginseng, tronadora, chromium, and alpha-lipoic acid. The popularity of these products varies among people of different ethnicities. Nopal is the most commonly used herbal hypoglycemic among persons of Mexican descent. Karela is more commonly used by persons from Asian countries. Some of these agents have gained universal appeal. For a select number of products, studies have revealed single or multiple mechanisms of action. For several of these, high soluble fiber content is a contributing factor. CONCLUSION: Based on the available evidence, several natural products in common use can lower blood glucose in patients with diabetes. Commonly used natural products often have a long history of traditional use, and pharmacists who have a stronger understanding of these products are better positioned to counsel patients on their appropriate use. PMID- 11926666 TI - New drugs of 2001. AB - OBJECTIVE: To provide information regarding the most important properties of the new therapeutic agents marketed in 2001. DATA SOURCES: Published studies, drug information reference sources, and product labeling. DATA SYNTHESIS: In 2001, 26 new therapeutic agents were marketed. The indications and information on dosage and administration for the new agents are reviewed, as are the most important pharmacokinetic properties, adverse events, drug interactions, and other precautions. Practical considerations for the use of the new agents are also discussed. Where possible, the properties of the new drugs are compared with those of older drugs marketed for the same indications. CONCLUSION: A number of the new therapeutic agents marketed in 2001 have important advantages over older medications. An understanding of the properties of these agents is important if the pharmacist is to effectively counsel patients about their use and to serve as a valuable source of information for other health professionals regarding these drugs. PMID- 11926667 TI - New OTC drugs and devices 2001: a selective review. AB - OBJECTIVES: To create a heightened awareness of two important issues that arose over the past year with respect to over-the-counter (OTC) drugs-the possible switching of nonsedating second-generation antihistamines to OTC status and the continuing need for pharmacists to learn about dietary supplements and herbal products-and to educate pharmacists about newly introduced nonprescription products, devices, and diagnostic products. DATA SOURCES: Recently published clinical and pharmaceutical industry literature. DATA SYNTHESIS: In a historical precedent, a third party insurer rather than a pharmaceutical company is attempting to force the switching of several prescription nonsedating antihistamines to OTC status. In addition, several developments related to the manufacture and use of dietary supplements have created concern for consumers and the pharmacists who advise them. It is critical for pharmacists to become knowledgeable about dietary supplements so they can appropriately counsel patients on the benefits and risks associated with supplement selection and use. In addition, numerous nonprescription products, including home diagnostic products and accessories, will continue to become available on the U.S. market. It is important that pharmacists become as knowledgeable as possible about these and be capable of educating the consuming public about their appropriate use. CONCLUSION: Patients will continue to place an increasing emphasis on self-care. To assist them, pharmacists must remain up-to-date on trends and have a balanced understanding of new nonprescription products and dietary supplements. The new nonprescription medications and diagnostic products discussed in this review article represent valuable additions to the growing array of self-care products. PMID- 11926668 TI - Use of herbal products for diabetes by Latinos. PMID- 11926670 TI - Counseling patients about sexual dysfunction. PMID- 11926669 TI - Patients' perceptions of independent community pharmacists. PMID- 11926671 TI - Omalizumab: a novel monoclonal antibody for treatment of allergic disease. PMID- 11926672 TI - Virtual panel provides the pharmaceutical industry fast, cost-effective answers. PMID- 11926673 TI - William Procter Jr. (1817-1874). PMID- 11926674 TI - Influence of a substance-abuse-prevention curriculum on violence-related behavior. AB - OBJECTIVE: To test the impact of a school-based substance-abuse-prevention program, Project Towards No Drug Abuse (TND), on risk for violence. METHODS: Logistic regression analyses tested whether victimization, perpetration, or weapon carrying differed for intervention students relative to control students within a sample of 850 continuation high school students followed over 12 months. RESULTS: We observed a higher risk for victimization (OR=1.57) among male control students. No intervention effect was observed for female students or for perpetration among males. CONCLUSION: The findings provide limited support for a generalization of TND's preventive effect. PMID- 11926676 TI - Impact of HIV test counseling on college students' sexual beliefs and behaviors. AB - OBJECTIVE: To compare the health beliefs and risky sexual behaviors of college students who did and did not seek HlV testing. METHODS: A survey was administered, and testing sessions were content analyzed. RESULTS: There were no significant differences in health beliefs, but students surveyed perceived themselves to be invulnerable to HIV/AIDS despite unsafe sexual behaviors. Sexual behaviors varied with students who sought HIV testing practicing more unsafe vaginal and oral intercourse. Assessment of the intervention suggested that counseling sessions were influential in persuading students to rethink their health beliefs and change their risky sexual behaviors. CONCLUSION: Testing can be used as both a prevention tool and a way to determine college students' HIV status. PMID- 11926675 TI - Intrinsic and extrinsic motivation and intention to breast-feed. AB - OBJECTIVE: To examine the feasibility of using the cognitive evaluation theory to examine pregnant women's intention to breast-feed. METHODS: A questionnaire designed to measure intrinsic and extrinsic motivation was administered to 228 pregnant women. RESULTS: Results provide evidence for reliability and validity of the revised instrument in this population. A factor analysis suggests the instrument measures 2 types of intrinsic motivation, one type of extrinsic motivation, and motivation related to the baby. The instrument distinguished differences in motivation between women who intend to breast-feed and those who intend to formula feed. CONCLUSION: This study helps elucidate motivational factors involved in infant-feeding decisions. PMID- 11926677 TI - Theory-based development and testing of an adolescent tobacco-use awareness program. AB - OBJECTIVE: To develop and disseminate an effective tobacco awareness and cessation program. METHODS: Self-report data from youth (n=1,601) and implementation data from site facilitators (n=16) provided the basis for the trial program evaluation. RESULTS: Approximately 146 total classes and 1,601 youth were served. Significantly (p<.01) more participants were in the action stage of change at the end of the program. At follow-up, 40% of the participants indicated they were tobacco free. CONCLUSION: The multiple iterative steps in program development were described. A theoretically based program, like the ATCP, is well worth exploring as a public health intervention. PMID- 11926678 TI - Predicting aerobic versus resistance exercise using the theory of planned behavior. AB - OBJECTIVE: To test the theory of planned behavior (TPB) in aerobic versus resistance training in a prospective design. Relationships between TPB variables, extroversion, and perceived health were examined. METHODS: College students (210) completed an initial measurement and a 3-month follow-up assessment. Reasons for exercising were assessed. RESULTS: TPB variables, extroversion, and perceived health collectively accounted for substantial variance in aerobic (19%) and resistance exercise (40%). Reasons for exercise included physical, psychological, and social concerns. CONCLUSION: Differences in the predictive validity of model constructs suggest potential differential intervention foci for aerobic versus resistance exercise. PMID- 11926679 TI - Determinants of physical activity in middle school children. AB - OBJECTIVE: To evaluate the theory of reasoned action (TRA) and planned behavior (TPB) in predicting moderate-to-vigorous physical activity (MVPA) in sixth-grade youth. METHODS: One hundred ninety-eight students completed a questionnaire measuring attitudes, subjective norms, perceived behavioral control, and intentions to be active. MVPA was measured using the CSA 7,164 accelerometer. RESULTS: Although demonstrating an acceptable fit, the TRA and TPB accounted for only a small percentage of the variance in MVPA. In support of the TPB, the addition of control perceptions to the reasoned action model added to the prediction of intentions and MVPA. CONCLUSION: Within our sample of sixth graders, the utility of the TRA or TPB as a framework for activity interventions appears to be limited. PMID- 11926680 TI - How cross-fostered chimpanzees (Pan troglodytes) initiate and maintain conversations. AB - This study systematically sampled typical attention-getting sounds and sign language conversations between each of 4 originally cross-fostered chimpanzees (Pan troglodytes), still living freely, but now in a laboratory setting, and a familiar human interlocutor. Videotape records showed that when they encountered a human interlocutor sitting alone at his desk with his back turned to them, the cross-fosterlings either left the scene or made attention-getting sounds. The only signs they made to the interlocutor's back were noisy signs. When the human turned and faced them, the chimpanzees promptly signed to him (98% of the time) and rarely made any sounds during the ensuing signed conversations. Under systematic experimental conditions, the signed responses of the chimpanzees were appropriate to the conversational styles of the human interlocutor, confirming daily field observations. PMID- 11926681 TI - Dogs' (Canis familiaris) responsiveness to human pointing gestures. AB - In a series of 3 experiments, dogs (Canis familiaris) were presented with variations of the human pointing gesture: gestures with reversed direction of movement, cross-pointing, and different arm extensions. Dogs performed at above chance level if they could see the hand (and index finger) protruding from the human body contour. If these minimum requirements were not accessible, dogs still could rely on the body position of the signaler. The direction of movement of the pointing arm did not influence the performance. In summary, these observations suggest that dogs are able to rely on relatively novel gestural forms of the human communicative pointing gesture and that they are able to comprehend to some extent the referential nature of human pointing. PMID- 11926682 TI - Use of experimenter-given cues in visual co-orienting and in an object-choice task by a new world monkey species, cotton top tamarins (Saguinus oedipus). AB - Two methods assessed the use of experimenter-given directional cues by a New World monkey species, cotton top tamarins (Saguinus oedipus). Experiment 1 used cues to elicit visual co-orienting toward distal objects. Experiment 2 used cues to generate responses in an object-choice task. Although there were strong positive correlations between monkey pairs to co-orient, visual co-orienting with a human experimenter occurred at a low frequency to distal objects. Human hand pointing cues generated more visual co-orienting than did eye gaze to distal objects. Significant accurate choices of baited cups occurred with human point and tap cues and human look cues. Results highlight the importance of head and body orientation to induce shared attention in cotton top tamarins, both in a task that involved food getting and a task that did not. PMID- 11926683 TI - Social transmission and memory of food preferences in pine voles (Microtus pinetorum). AB - The social transmission of food preferences is affected by factors including the length of time a preference is maintained. The authors investigated the social transmission of food preferences in pine voles (Microtus pinetorum) and whether food items had to be present for memory to persist. A demonstrator vole was fed cocoa-flavored food, a nonpreferred food. After interacting with a demonstrator vole, observer voles preferred cocoa-flavored food as compared with either naive voles that had not interacted with demonstrators or voles that had been exposed to odors. Observers retained this preference for 72 hr after interacting with an observer whether or not flavored foods were present. The ability to learn which food items are palatable and safe may benefit a generalist herbivore that has faced selection to avoid toxic baits. PMID- 11926685 TI - Preference for novel flavors in adult golden hamsters (Mesocricetus auratus). AB - Although animals generally prefer to eat foods with familiar rather than unfamiliar flavors, adult golden hamsters (Mesocricetus auratus) were found to do the opposite. After having prolonged exposure to a food with a particular flavor, hamsters were allowed to select between the food with the familiar flavor and the same food with a novel flavor. Hamsters consistently ate more of the food with the novel flavor, and this preference was long-lasting and resistant to extinction. Furthermore, the novelty effect was robust, being manifested in both sexes and under a variety of experimental circumstances. In contrast, rats tested under identical conditions consistently preferred the food with the familiar flavor. The origins of the novelty effect in hamsters remain to be determined. PMID- 11926684 TI - The relationship between problem solving and inhibitory control: cotton-top tamarin (Saguinus oedipus) performance on a reversed contingency task. AB - To explore the relationship between problem solving and inhibitory control, the authors present 4 experiments on cotton-top tamarins (Saguinus oedipus) using a reverse-reward contingency task. In Experiment 1, 1 group of tamarins was given a choice between a small and a large quantity of food. Whichever quantity the tamarins reached for first, they received the alternative. The tamarins consistently picked the larger quantity, thereby receiving the smaller. A 2nd group of tamarins was given the same task, except that if they reached for the larger quantity of food, they received nothing. The tamarins continued to pick the larger quantity, even though this resulted in no food. In addition, most of the tamarins continued to pick the larger quantity even when the food payoff for choosing the smaller quantity was increased (Experiment 2) or when the visual salience of the food was reduced (Experiment 3). Experiment 4 was based on the finding that chimpanzees (Pan troglodytes) that have been trained on the concept of number can solve the reversed contingency task if the food is replaced by Arabic numerals. With the help of a color association, and a higher cost incurred by picking the color associated with 3 food items, the tamarins learned to pick the color associated with 1 food item. These results are compared with those obtained from studies of other primate species, highlighting the importance of comparative studies of problem solving that use comparable methods. PMID- 11926686 TI - Social contacts and production of 50-kHz short ultrasonic calls in adult rats. AB - The goal of the study was to provide evidence that the production of 50-kHz calls by adult rats is driven by potential or direct social contacts. The calls have been studied during daily visits to a cage by single or paired rats. Repeated exposure of rats to the cage frequently visited by other rats or direct contact between rats significantly increased the number of 50-kHz calls. The increase in production of 50-kHz calls was reduced by 78% after intrapreoptic-anterior hypothalamic injection of MK-801, an N-methyl-D-aspartate-type glutamate receptor antagonist. Calls emitted in all situations had a similar acoustic profile. It was found that 50-kHz calls were produced in anticipation of, and/or during, direct social contacts among adult rats and were predominantly initiated by olfactory stimuli. The calls seem to express an appetitive behavioral state in which the central glutamatergic mechanism is implicated. PMID- 11926688 TI - Research in action: a summary of the 2001 American Osteopathic Association Research Conference. PMID- 11926689 TI - Osteopathic philosophy must be the foundation of osteopathic medical education. PMID- 11926687 TI - Acute hunger of rat pups elicits increased kyphotic nursing and shorter intervals between nursing bouts: implications for changes in nursing with time postpartum. AB - Earlier findings, based on limited behavioral observations, indicate that nursing behavior in rats declines dramatically in duration over time postpartum-despite increasing ingestion of milk by rat pups to meet their growth and metabolic needs although hungry pups elicit more nursing than do well-nourished pups. The authors compared the nursing pattern in detail for 6 hr on Days 7 and 14 and induced hunger in pups acutely with mammary-duct-ligated dams unable to provide milk. Compared with Day 7, on Day 14, supine nursing and the interval between nursing bouts increased, whereas hovering over pups and kyphotic nursing decreased. When pups were increasingly hungry, these age-related changes were counteracted. Thus, the ingestive motivation of pups largely regulates the nursing pattem over time. PMID- 11926690 TI - Is pediatric attendance necessary for all cesarean sections? AB - The authors analyzed mode of delivery and resuscitation rates in a group of 1410 newborns. Comparisons were made between vaginal and cesarean deliveries, and these were further stratified with regard to whether cesarean section was performed as a routine elective procedure or whether an indication existed that necessitated operative delivery of the child (nonelective). A significantly increased risk for resuscitation was found in the cesarean section group overall when compared to vaginal deliveries. The risk of resuscitation after repeat elective cesarean section was low, and this risk increased significantly in the nonelective cesarean delivery group. As the risk of resuscitation after elective repeat cesarean sections is not significantly increased, it may not be necessary that a pediatrician be present at these deliveries. PMID- 11926691 TI - The "iliacus test": new information for the evaluation of hip extension dysfunction. AB - This study confirms the clinical value of investigating the "iliacus complex" during evaluations of the low back. A new "iliacus test" isolates this iliacus complex component of limited hip extension. Designed for a single joint, the test isolates motion across the hip joint. Study results include the following: (1) in a comparison with the clinical standard, the Thomas test, data show that the two tests are significantly different in an asymptomatic population between the ages of 18 and 35 years; (2) with the exception of the standard Thomas test, the data show no statistical differences in range of motion when comparing the left side with the right side; (3) examiner-added, end-range pressure for assessment of range of motion when compared with the standard gravity-dependent end range of motion used in the Thomas test yields valuable new information; and (4) data provide a basis for population norms for each test--Thomas and iliacus--in gravity-dependent and examiner-produced tissue-feel end ranges. PMID- 11926692 TI - Evaluation of osteopathic manipulative treatment training by practicing physicians in Ohio. AB - The authors mailed a survey designed to evaluate beliefs about osteopathic manipulative treatment (OMT) training to the 2318 osteopathic physicians registered with the Ohio Osteopathic Association. Responses were received from 871 osteopathic physicians (response rate, 38%). Fifty-three percent of the respondents had used OMT with patients at least once during the week before the survey. With regard to OMT training, 60% rated their experience during medical school as acceptable; during postgraduate training the acceptable rating dropped to 9%. Osteopathic manipulative treatment training through continuing medical education programs was rated as acceptable by 26% who had participated in these programs. Forty percent of the respondents reported that they were practicing less OMT now than when they originally entered practice, while 20% reported using OMT procedures more often. No significant correlation was observed between OMT training satisfaction during medical school and current use of OMT. However, a strong negative correlation was observed between satisfaction with postgraduate OMT training and OMT use. This survey did not detect any association between year of graduation and use of OMT. PMID- 11926693 TI - Quality of life in referred patients presenting to a specialty clinic for osteopathic manipulative treatment. AB - Previous research has found that patients of osteopathic physicians tend to report poorer general health perceptions than persons in the general population or than patients of allopathic physicians. Quality of life and level of healthcare satisfaction in patients referred to a specialty clinic for osteopathic manipulative treatment (OMT) at a college of osteopathic medicine were measured in 1997. Data from the Medical Outcomes Study 36-Item Short Form (SF-36) were used to compute standardized scores in the following eight health scales: physical functioning, role limitations because of physical problems, bodily pain, general health perceptions, vitality, social functioning, role limitations because of emotional problems, and mental health. There were 185 patients who returned the survey (mean response rate, 90%), including 22 new and 163 established patients. Patients reported poorer health than the general population on all eight scales (P < .001). Patients frequently reported poorer quality of life than referents with hypertension, congestive heart failure, type 2 diabetes mellitus, recent acute myocardial infarction, or clinical depression. More than 97% of established patients were satisfied or very satisfied with the healthcare received at the clinic. This study suggests that referred patients presenting to osteopathic physicians for OMT may have poorer quality of life than is generally recognized when relying only on traditional diagnostic approaches. Early detection and treatment of musculoskeletal conditions may be important factors in preventing chronicity and its impact on quality of life. PMID- 11926694 TI - Current threats to osteopathic graduate medical education. AB - The Balanced Budget Act of 1997 and continuing changes put into place by the Educational Commission on Foreign Medical Graduates (ECFMG) are altering the environment for graduate medical education (GME) in ways that threaten osteopathic graduate medical education in particular. Hospital revenue is decreasing due to declines in Medicare GME and patient-care reimbursements. The new 3-year rolling average methodology for counting "house staff" makes it likely that unfilled positions will be eliminated. With osteopathic GME positions unfilled and financial resources decreasing, osteopathic medical programs may shrink further. Additionally, the ECFMG has put into place policies that may restrict the number of international medical graduates entering the United States. Approximately 25% of all allopathic GME positions in the United States are filled by international medical graduates. If this applicant pool decreases, allopathic medical programs may turn to osteopathic medical graduates as the only other available pool of individuals to fill program positions. At a time when allopathic internship positions are already unfilled and 30% of osteopathic medical graduates enter allopathic first-year programs, further inroads by allopathic programs could severely impact osteopathic GME efforts. PMID- 11926695 TI - Efficacy of a proprietary ibuprofen gel in soft tissue injuries: a randomised, double-blind, placebo-controlled study. AB - The efficacy of a novel, proprietary topical formulation of ibuprofen 5% gel (Ibugel) was evaluated in a placebo-controlled study in patients with soft tissue injuries. Patients received either active gel (n=40) or placebo gel (n=41) for a maximum of seven days. Pain and interference with physical activity were assessed daily using visual analogue scales. There was a significant difference (p<0.001) in favour of active treatment for the time to achieve clinically meaningful reduction in pain. By day 7, 75% of patients in the active gel group had a clinically meaningful reduction of pain compared with 39% of patients who received placebo. Despite differences between study centres, the data for interference with physical activity also showed an advantage for active treatment. By day 7, 79% of patients in the active gel group had a clinically meaningful reduction in interference with physical activity, compared with 44% of patients who received placebo. PMID- 11926696 TI - 9. Arm lymphoedema following breast cancer treatment. AB - Lymphoedema of the upper limb can be a devastating consequence of breast cancer treatment. A wide range of incidence has been reported but the generally accepted rate is around 12%. The exact pathophysiological mechanisms remain unclear. Agreed working definitions are required in order to standardise future research in this area. The psychosocial importance and management is fundamental in limiting the impact of this phenomenon. Furthermore, it remains that the primary cause is dissection of the axilla. Therefore, active investigation into potential tools that may eliminate or reduce the extent of dissection without compromising prognosis is warranted. PMID- 11926697 TI - 10. Sentinel node biopsy in primary breast cancer. AB - Sentinel node biopsy is a minimally invasive but accurate way of staging the axilla such that as many as 50% of women with primary breast cancer could avoid axillary clearance and the morbidity this carries with it. The methodology has yet to be perfected but identification of the sentinel node(s) with either dye or isotope, or with a combination of these, is a robust technique that is quick to learn and correctly predicts the status of the rest of the axilla in significantly more than 90% of patients. Improvements in the methodology and in patient selection will inevitably make sentinel node biopsy even more accurate. Studies to demonstrate the survival impact of replacing axillary clearance with sentinel node biopsy and selective axillary node treatment are required and are in their early stages. PMID- 11926698 TI - Selective cholesterol absorption inhibition: a novel strategy in lipid-lowering management. AB - Many individuals throughout Europe have risk factors for coronary heart disease (CHD) and are non-compliant with recommended treatments, despite guidelines for the reduction of low-density lipoprotein cholesterol (LDL-C) and the prevention of CHD. Significant numbers who should receive pharmacotherapy for hypercholesterolaemia do not, and one-third of treated patients do not achieve recommended target LDL-C levels. Optimum doses of statins, which have demonstrated undisputed efficacy in the treatment of hypercholesterolaemia in clinical trials, are seldom used; the inconvenience of dosage adjustments and safety concerns, particularly myalgia, may constitute obstacles to their optimal use for LDL-C reduction in clinical practice. Ezetimibe is the first selective cholesterol absorption inhibitor that has demonstrated clinical benefits when used as either monotherapy or in combination with other lipid-modifying agents. PMID- 11926699 TI - Dipyridamole may be used safely in patients with ischaemic heart disease. AB - It is thought that up to 50% of patients with cerebrovascular disease will have concurrent ischaemic heart disease. Dipyridamole co-formulated with aspirin has been shown to increase the relative reduction in risk of second stroke in patients with prior stroke/transient ischaemic attack beyond that obtaining with aspirin alone. We have sought to resolve the question of whether dipyridamole treatment increases the risk of cardiac adverse events in patients with co existing ischaemic heart disease. The published literature, periodic safety update reports, the randomised controlled trials of antiplatelet agents in stroke prevention and those including dipyridamole in cardiovascular indications, have been reviewed and analysed. The early reports of serious adverse cardiac effect attributable to dipyridamole occurred in patients with severe coronary artery disease using dipyridamole as a stress test adjunct to cardiac imaging. The randomised controlled trials databases show no evidence of mortality and only isolated cases of significant cardiac morbidity attributable to dipyridamole at recommended oral doses in patients with ischaemic heart disease. We conclude that patients with cerebrovascular and mild to moderate concomitant ischaemic heart disease may be treated safely with dipyridamole for the secondary prevention of stroke. PMID- 11926700 TI - An overview of the pharmacology, efficacy, safety and cost-effectiveness of lansoprazole. AB - Lansoprazole is a proton pump inhibitor that reduces gastric acid secretion in a dose-dependent manner via inhibition of H+/K+-adenosine triphosphatase in gastric parietal cells. It also exhibits antibacterial activity against Helicobacter pylori in vitro. During almost 10 years of clinical use, lansoprazole has proved effective and well tolerated in a wide range of acid-related disorders, including gastro-oesophageal reflux disease (GORD), duodenal ulcers, gastric ulcers, non steroidal anti-inflammatory drug-related ulcers, as well as non-ulcer dyspepsia and acid hypersecretion. It is also used, in combination with antibiotics, for H. pylori eradication. In the above indications, lansoprazole has generally proved to be superior to the histamine H2-receptor antagonists, and is at least as effective as the other currently available proton pump inhibitors. This review aims to evaluate the pharmacology, efficacy, safety and cost-effectiveness of lansoprazole in acid-related disorders, with particular emphasis on its use in GORD and H. pylori eradication regimens. PMID- 11926701 TI - The potential of mifepristone (RU486) as a female contraceptive drug. AB - This article reviews the development of mifepristone (RU486) as a female contraceptive drug. Mifepristone is an orally active compound with nearly 40% bioavailability after first pass effect. The steady plasma level of mifepristone ranges from 65 nmol/l with 1 mg/day to 1 micromol/l with 10 mg/day and reaches 2.5 micromol/l, 4.5 micromol/l and 5.4 micromol/l with mifepristone 50 mg, 100 mg and 200 mg daily, respectively, over the treatment period. Inhibition of ovulation may be achieved at serum mifepristone concentration of 232.7 nmol/l. Mifepristone appears to antagonise progesterone at the pituitary level to suppress gonadotropin and steroid hormone secretion rather than to act primarily on the hypothalamus to delay or inhibit ovulation. In fact, the endometrium is most sensitive to mifepristone. Low-dose mifepristone impairs luteal phase endometrial development and receptivity by altering endometrial parakine, cytokine and enzyme activity. Thus, low-dose mifepristone can significantly reduce the rate of conception without inhibiting ovulation. However, further research is needed to standardise the dose and dose-schedule to achieve the desired efficacy of low-dose mifepristone for routine clinical use with minimal or no side-effects. PMID- 11926702 TI - Adverse cutaneous reaction to naproxen: a missed diagnosis fortuitously resolved. AB - A 91-year-old man presented with a one-month history of swollen legs and dyspnoea. He also complained of a 16-year chronic rash that had caused him considerable morbidity and cosmetic problems. He had been taking naproxen for several years for osteoarthritis. Clinical examination demonstrated bilateral pitting oedema of the leg and a widespread excoriated rash that affected most of his body. A clinical diagnosis of fluid retention secondary to naproxen was made. Stopping naproxen led not only to the resolution of his leg oedema and dyspnoea, but also to the prompt and complete remission of the rash. Reviewing the patient's history revealed that the rash had almost certainly first appeared when naproxen was started many years previously. PMID- 11926703 TI - Non-invasive imaging methods before fine-needle aspiration in the diagnosis of cervical masses. AB - A carotid body tumour is a vascular tumour usually located at or around the bifurcation of the carotid artery and originating from the tunica adventitia. Carotid body tumours, which are generally non-functional and asymptomatic, grow slowly, which is why diagnosis can be delayed. Because of the vascular structures, fine-needle aspiration biopsy may be dangerous and impractical. Efficient and reliable methods of diagnosis, such as colour Doppler ultrasound, magnetic resonance imaging, computerised tomography and arteriography, can be useful. We report on a 58-year-old woman with diarrhoea, flushing and a cervical, hard, non-tender, fixed mass. An unsuccessful diagnostic fine-needle aspiration biopsy was performed after palpation of the cervical mass. A carotid body tumour was finally diagnosed. We concluded that non-invasive imaging methods should be evaluated before fine-needle aspiration in cervical masses. It should also be kept in mind that asymptomatic cervical masses may be related to non-local symptoms. PMID- 11926705 TI - Sudden permanent hearing loss following anterior inferior cerebellar artery infarction. AB - Anterior inferior cerebellar artery infarction with the only sequel being a permanent unilateral hearing loss is described. The damage was confirmed by magnetic resonance imaging. Hearing loss of vascular cause may be more common and permanent than realised, and missed if the other neurological deficits have resolved. PMID- 11926704 TI - Successful pregnancy following gonadotropin therapy in a patient with hypogonadotropic hypogonadism resulting from craniopharyngioma. AB - The authors report a rare case of a patient with panhypopituitarism who became pregnant by gonadotropin therapy and gave birth to a healthy baby. A brain tumour and/or the surgical resection of a brain tumour occasionally results in pituitary dysfunction. An 18-year-old Japanese patient developed hypogonadotropic secondary amenorrhoea because of a craniopharyngioma, which was surgically removed. The patient came to us, and peripheral blood was collected every 15 minutes for four hours. The levels of luteinising hormone (LH) and follicle-stimulating hormone (FSH) were measured. Results showed that LH and FSH levels were very low and did not fluctuate. Several years later, the patient complained of infertility, and treatment with human menopausal gonadotropin (hMG) and human chorionic gonadotropin (hCG) was started. The therapy was repeated for several cycles, but she did not conceive, so hMG-hCG therapy combined with conjugated oestrogen administration was started. The patient became pregnant at the seventh cycle of this combined therapy. She was not treated with supplementary growth hormone. PMID- 11926706 TI - Bleeding angiodysplasia: should we concentrate more on the aortic valve than on the bowel? AB - A case of bleeding from angiodysplasia in association with aortic stenosis is presented. The particular interest in this patient is that the bleeding ceased immediately after the insertion of a bioprosthetic valve and recurred when the valve restenosed. Furthermore, the bleeding again promptly resolved when the valve was exchanged for a metallic prosthesis despite long-term anticoagulation. The implications of these observations are discussed. PMID- 11926707 TI - The oxygen-air equation. PMID- 11926708 TI - Sildenafil (Viagra): new data, new confidence in treating erectile dysfunction in the cardiovascular patient. PMID- 11926710 TI - Acceptable agreement between tympanic and rectal temperature in acute stroke patients. AB - Major concerns remain about the reliability of tympanic thermometers. The aim of this study was to investigate the agreement between rectal and tympanic thermometry and compare this with the repeatability of each method in patients with acute stroke. Tympanic temperature readings were from 0.7 degrees C below to 0.8 degrees C above rectal readings. As to repeatability, we found second rectal readings to be between 0.3 degrees C below and 0.4 degrees C above first readings, and second tympanic readings to be between 0.4 degrees C below and 0.5 degrees C above first readings. The agreement between the two methods is acceptable in clinical practice. Tympanic thermometry is useful in serial measurements, whereas, for a single accurate measurement, rectal mercury thermometry remains the method of choice. PMID- 11926709 TI - A cost-effectiveness evaluation of amlodipine usage in patients with coronary artery disease in Sweden. AB - The objective of this analysis was to calculate the cost-effectiveness of amlodipine therapy in patients with coronary artery disease in Sweden. It is hypothesised that treatment with amlodipine will have an impact on overall cardiovascular disease treatment costs, resulting in a positive cost effectiveness profile. A Markov cohort simulation model was constructed to simulate event-related and procedure-related health economic outcomes of coronary artery disease populations on amlodipine versus those on placebo. Patient level data from the Prospective Evaluation of the Vascular Effects of Norvasc Trial was used to populate the model. The total number of adverse cardiovascular clinical outcomes experienced over a three-year period was lower for patient on amlodipine than for those on placebo. The rate of hospitalisation per patient due to angina, coronary artery bypass graft, percutaneous transluminal coronary angioplasty, congestive heart failure, and myocardial infarction in the placebo cohort was 64.7%, while the rate in the amlodipine cohort was 46.9%. The cost per patient was Swedish kroner (SEK)26,600 for amlodipine patients and SEK27,400 for placebo patients. The use of amlodipine resulted in improved clinical outcomes as well as a slight savings in cost over a three-year period. PMID- 11926711 TI - Comparison of non-invasive ventilation and standard medical therapy in acute hypercapnic respiratory failure: a randomised controlled study at a tertiary health centre in SE Turkey. AB - This study was designed in a tertiary health centre in south-eastern Turkey to compare the effectiveness of non-invasive ventilation (NIV) plus standard medical therapy (ST) to ST alone, in acute hypercapnic respiratory failure (AHRF) due to chronic obstructive pulmonary disease (COPD) exacerbation. Thirty-four consecutive patients were randomly assigned to receive either NIV plus ST or ST alone. NIV was applied with a simple non-invasive ventilator through a full face mask in the general ward. Initial settings for inspiratory and expiratory positive airway pressures were 9 cm HO2, and 3 cm HO2, respectively. We observed statistically significant improvements in the first hour of NIV regarding respiratory rate (p<0.001), pH (p<0.05), PaO2 (p<0.05), and PaCO2 (p<0.001). PaO2 (p<0.05) showed significant improvement only in the first hour of ST. The intubation rate and duration of hospitalisation in the NIV group were significantly shorter than those in the ST group (p<0.05). We conclude that NIV provides adjunctive therapeutic benefits compared with ST alone, and should be the choice of first step treatment in the AHRF due to COPD exacerbation in the appropriate setting and in selected patients. PMID- 11926712 TI - Understanding patient perceptions of asthma: results of the Asthma Control and Expectations (ACE) survey. AB - In spite of support among UK healthcare professionals for asthma guidelines, studies continue to show that many patients fail to reach the suggested management goals. Patient expectations and poor communication may be factors in this failure. This survey assessed patients' asthma control, expectations in respect of asthma and communication with healthcare professionals. A structured questionnaire, designed for self-completion, was developed and distributed to asthma patients at participating pharmacies. A total of 1031 questionnaires were returned. Most of the respondents, even among those feeling well, reported lifestyle restrictions because of asthma. These restrictions were not generally discussed with healthcare professionals. One-third did not perceive the benefits of inhaled corticosteroids. Low expectation may be a major contributor to the poor control seen in this survey. The failure to discuss lifestyle restrictions and symptom levels with healthcare professionals gives little scope for recommendation of appropriate and adequate treatment. Improved communication between healthcare professionals and patients may help such understanding and raise patient expectations. PMID- 11926713 TI - Multidose flurbiprofen 8.75 mg lozenges in the treatment of sore throat: a randomised, double-blind, placebo-controlled study in UK general practice centres. AB - The flurbiprofen 8.75 mg lozenge is a novel formulation that combines a demulcent effect with the analgesic activity of a non-steroidal anti-inflammatory drug. Previous controlled clinical studies have demonstrated the single- and multi-dose efficacy of these lozenges over placebo. The current study reflected the treatment of sore throat in general practice, investigating multiple dose efficacy where patients also had access to concomitant antibiotics and rescue medication. The efficacy of flurbiprofen 8.75 mg lozenge over placebo was confirmed: there was a significant difference in pain relief obtained from flurbiprofen 8.75 mg versus placebo, along with a significant reduction of difficulty in swallowing from the time of first assessment and significantly greater reductions in throat soreness and difficulty in swallowing throughout the study period. Additionally, significant benefit over placebo was demonstrated where concomitant antibiotic use was introduced, indicating that flurbiprofen 8.75 mg lozenges can be co-administered when antibiotic therapy is appropriate. No significant safety issues were identified. PMID- 11926714 TI - Selective actions on sleep or anxiety by exploiting GABA-A/benzodiazepine receptor subtypes. AB - The next generation of anxiolytics may be subtype-selective partial agonists at GABA-A receptors. By exploiting new psychopharmacologic principles, such as the targeting of receptor subtypes that have unique subunits and the partial activation of these subtypes with stabilizers, we may eventually have anxiolytics that are rapid acting, but without sedation, amnesia, or dependence. PMID- 11926715 TI - Combining bupropion SR with venlafaxine, paroxetine, or fluoxetine: a preliminary report on pharmacokinetic, therapeutic, and sexual dysfunction effects. AB - BACKGROUND: This study was designed to evaluate the effect of combining bupropion sustained release (SR) with venlafaxine, paroxetine, or fluoxetine in patients who reported unacceptable sexual dysfunction when treated with monotherapy with the latter 3 agents. METHOD: Following a minimum of 6 weeks of antidepressant treatment with a selective serotonin reuptake inhibitor (SSRI) or venlafaxine (a serotonin-norepinephrine reuptake inhibitor), eligible subjects received a further 8 weeks of monitored combination therapy with bupropion SR at a dose of 150 mg/day with no alterations to index antidepressant dosing. RESULTS: There was a clinically significant benefit in 14 (78%) of 18 partial responders or nonresponders, and 33% (N = 6) achieved a full response (chi2= 8.06, df = 2, p = .017). Sexual dysfunction, particularly a decrease in orgasmic delay, was also significantly improved with combination therapy (men: paired t = -2.1, df = 6, p = .08; women: paired t = -3.0, df = 7, p = .02). Plasma monitoring of drugs and their metabolites revealed a statistically significant increase in venlafaxine levels (F = 6.89, df = 4,24; p = .001) accompanied by a decrease in O desmethylvenlafaxine (F = 14.26; df = 4,24; p < .0005) during combined treatment with bupropion SR. There were no statistically significant changes in plasma levels of SSRIs (paroxetine and fluoxetine) during the trial. CONCLUSION: Bupropion had an effect on the pharmacokinetics of venlafaxine but not those of the SSRIs. Further investigation of combination treatments under randomized, double-blind conditions is recommended. PMID- 11926716 TI - Major depressive disorder and axis I diagnostic comorbidity. AB - BACKGROUND: Recognition of comorbid conditions in patients presenting for the treatment of depression is clinically important because the presence of other disorders can influence treatment planning. In the present study, we examined the frequency of diagnostic comorbidity in psychiatric outpatients presenting for treatment of nonbipolar major depressive disorder (MDD) and patients' desire for treatment for the comorbid disorders. METHOD: Four hundred seventy-nine psychiatric outpatients with DSM-IV nonbipolar MDD were evaluated with a modified version of the Structured Clinical Interview for DSM-IV. RESULTS: Excluding nicotine dependence, at the time of the evaluation 64.1% (N = 307) of the patients met criteria for at least 1 of the 23 specific Axis I disorders, and more than one third (36.7%, N = 176) had 2 or more disorders. Anxiety disorders, as a group, were the most frequent current comorbid disorders (56.8%), and social phobia was the most frequent individual disorder. Including subthreshold conditions, the percentage of patients with at least 1 disorder increased to 73.5%. When the scope of assessment was expanded to include nicotine dependence, nicotine dependence was the most frequent lifetime individual disorder (38.2%) and the second most frequent current disorder (27.3%). There was considerable variability among the disorders regarding desire for treatment of the comorbid condition. CONCLUSION: The majority of nonbipolar depressed patients have a current comorbid disorder, especially an anxiety disorder, although the actual rate of comorbidity depends on the breadth of the assessment. PMID- 11926717 TI - An open-label trial of citalopram for major depression in patients with hepatitis C. AB - BACKGROUND: Hepatitis C affects nearly 4 million Americans. Depression is a common comorbid condition in this population and may be induced by interferon alfa, an approved treatment for hepatitis C. Depression is a major indicator for discontinuation of interferon therapy. This open-label study examines the effect of citalopram on measures of depression and quality of life and tests of liver function in subjects with hepatitis C and major depressive disorder. METHOD: Subjects were recruited by advertisement; those with DSM-IV major depressive disorder were included in the study. Subjects received citalopram for 8 weeks starting at 20 mg/day. Dosage adjustments were made as the physicians deemed clinically necessary. No dosages were increased prior to week 4 of the study. Hamilton Rating Scale for Depression (HAM-D) scores, Clinical Global Impressions Severity of Illness scale (CGI-S) scores, Medical Outcomes Study Short Form Health Survey (SF-36) ratings, Symptom Checklist-90-Revised (SCL-90-R) scores, and liver function tests were obtained at baseline, 4 weeks, and 8 weeks. RESULTS: A total of 15 patients (10 men, 5 women) participated in this study. The mean daily dose of citalopram at endpoint was 26.67 mg. Mean HAM-D scores decreased significantly with treatment (F = 36.3, df = 2,42; p = .0001). Thirteen of the 15 subjects demonstrated a clinical response, defined as a 50% or greater reduction in HAM-D scores. CGI-Severity of Illness scores also improved significantly (p = .0001). Subjects demonstrated statistically significant improvement (p < .05) on all of the SF-36 subscales. Statistically significant improvements (p < .05) were also demonstrated on all subscales of the SCL-90-R. Tests of liver function showed no significant worsening of aspartate aminotransferase, alanine aminotransferase, or gamma-glutamyltransferase levels. CONCLUSION: These results suggest that depression in patients with hepatitis C may be effectively and safely treated with citalopram. PMID- 11926718 TI - Fluoxetine versus placebo in posttraumatic stress disorder. AB - BACKGROUND: This study was designed to address the efficacy and tolerability of fluoxetine in patients with posttraumatic stress disorder (PTSD) as diagnosed using the Structured Clinical Interview for DSM-IV Axis I Disorders and the Clinician-Administered PTSD Scale (CAPS). The patient population included both civilians and combat veterans. METHOD: This was a double-blind, randomized, placebo-controlled study conducted in Europe, Israel, and South Africa, primarily in war-torn countries. Patients were predominantly male (81%) and white (91%), with 48% exposed to a combat-related traumatic episode. Patients were randomly assigned to 12 weeks of acute treatment with fluoxetine, 20 to 80 mg/day (N = 226), or placebo (N = 75). The primary efficacy measurement was the mean change from baseline in the Treatment Outcome PTSD rating scale (TOP-8) total score, which was analyzed using a repeated-measures analysis of variance. Secondary assessments included the CAPS, the Davidson Trauma Scale, the Clinical Global Impressions-Severity of Illness scale (CGI-S), the CGI-Improvement scale (CGI-I), the Montgomery-Asberg Depression Rating Scale (MADRS), the Hamilton Rating Scale for Anxiety (HAM-A), and the Hopkins 90-Item Symptom Checklist-Revised. RESULTS: Fluoxetine was associated with a greater improvement from baseline in total TOP-8 score than was placebo. This difference was statistically significant by week 6 of treatment (p < .001) through the end of the acute phase of the study (week 12; p = .006). Compared with placebo, fluoxetine was also associated with significantly greater improvement in CAPS total score as well as intrusive and hyperarousal subscores and in CGI-S, CGI-I, HAM-A, and MADRS scores (p < .05). The presence of dissociative symptoms at baseline appeared to be a predictor of high placebo response. The mean fluoxetine dose at endpoint was 57 mg. There were no clinically significant safety differences. CONCLUSION: Fluoxetine is effective and well tolerated in the treatment of PTSD. Most PTSD patients will respond satisfactorily at doses in the upper normal range for the usual antidepressant doses of fluoxetine. PMID- 11926719 TI - Correlates of overweight and obesity in 644 patients with bipolar disorder. AB - OBJECTIVE: Overweight and obesity are common clinical problems encountered in the treatment of bipolar disorder. We therefore assessed the prevalence and clinical correlates of overweight, obesity, and extreme obesity in 644 bipolar patients. METHOD: 644 outpatients with DSM-IV bipolar disorder in the Stanley Foundation Bipolar Treatment Outcomes Network were evaluated with structured diagnostic interviews and clinician- and self-administered questionnaires to determine bipolar disorder diagnoses, demographic and historical illness characteristics, comorbid Axis I diagnoses, medical histories, health habits, and body mass indices (BMMs). RESULTS: Fifty-eight percent of the patients with bipolar disorder were overweight, 21% were obese, and 5% were extremely obese. American patients had significantly higher mean (p < .0001) BMIs and significantly higher rates of obesity (p < .001) and extreme obesity (p < .001) than European patients. Significant associations (p < or = .001) were found between overweight, obesity. and extreme obesity and gender, age, income level, comorbid binge-eating disorder, hypertension, arthritis, diabetes mellitus, exercise habits, and coffee consumption. Current BMI and weight were each correlated with the number of weight gain-associated psychotropics to which patients had been exposed. Multinomial logistic regression (adjusted for site and eating disorder diagnosis and corrected for multiple comparisons) showed that (1) overweight was significantly associated with male gender and hypertension (p < .001), (2) obesity was significantly associated with hypertension (p < .001), and (3) extreme obesity was significantly associated with hypertension and arthritis (p < .001). CONCLUSION: Overweight, obesity, and extreme obesity were common in this group of bipolar patients, although it was unclear that their prevalence rates were truly elevated, because overweight and obesity are increasingly common public health problems among the general population. Correlates of overweight and obesity in bipolar disorder include patient and treatment variables such as gender, geographical location, comorbid binge-eating disorder, age, income level, degree of exposure to weight gain-associated psychotropics, medical disorders associated with obesity, and health habits. PMID- 11926720 TI - Normal P50 gating in children with autism. AB - BACKGROUND: An important characteristic of children with autism is their unusual reaction to stimuli, which may be related to problems in the filtering of sensory input. For this reason, sensory filtering was measured in children with autism using the P50 gating paradigm. METHOD: Twelve non-mentally retarded children with autism (i.e., having a DSM-IV diagnosis of either autistic disorder or pervasive developmental disorder not otherwise specified) and 11 healthy control children were tested for their ability to suppress P50, measured at the Cz electrode. RESULTS: No differences were found between the children with autism and the control children with regard to absolute P50 amplitudes and P50 suppression. CONCLUSION: The excitability of the neuronal substrate that causes P50 is normal in children with autism, as are the early, inhibitory processes related to P50 gating. These results distinguish between subjects with autism and subjects with schizophrenia, in whom sensory gating is abnormal. PMID- 11926721 TI - Carbamazepine augmentation for schizophrenia: how good is the evidence? AB - BACKGROUND: Augmentation strategies in schizophrenia treatment remain an important issue because despite the introduction of several new antipsychotics, many patients remain treatment resistant. The aim of this study was to undertake a systematic review and meta-analysis of the safety and efficacy of one frequently used adjunctive compound: carbamazepine. DATA SOURCES AND STUDY SELECTION: Randomized controlled trials comparing carbamazepine (as a sole or as an adjunctive compound) with placebo or no intervention in participants with schizophrenia or schizoaffective disorder were searched for by accessing 7 electronic databases, cross-referencing publications cited in pertinent studies, and contacting drug companies that manufacture carbamazepine. METHOD: The identified studies were independently inspected and their quality assessed by 2 reviewers. Because the study results were generally incompletely reported, original patient data were requested from the authors; data were received for 8 of the 10 randomized controlled trials included in the present analysis, allowing for a reanalysis of the primary data. Dichotomous variables were analyzed using the Mantel-Haenszel odds ratio and continuous data were analyzed using standardized mean differences, both specified with 95% confidence intervals. RESULTS: Ten studies (total N = 283 subjects) were included. Carbamazepine was not effective in preventing relapse in the only randomized controlled trial that compared carbamazepine monotherapy with placebo. Carbamazepine tended to be less effective than perphenazine in the only trial comparing carbamazepine with an antipsychotic. Although there was a trend indicating a benefit from carbamazepine as an adjunct to antipsychotics, this trend did not reach statistical significance. CONCLUSION: At present, this augmentation strategy cannot be recommended for routine use. The most promising targets for future trials are patients with excitement, aggression, and schizoaffective disorder bipolar type. PMID- 11926722 TI - Duloxetine in the treatment of major depressive disorder: a double-blind clinical trial. AB - BACKGROUND: Duloxetine hydrochloride, a dual reuptake inhibitor of serotonin and norepinephrine, was evaluated for therapeutic efficacy and safety/tolerability in the treatment of major depression. METHOD: In an 8-week multicenter, double blind, placebo-controlled study, 173 patients (aged 18-65 years) with DSM-IV major depressive disorder were randomly allocated to receive placebo (N = 70), duloxetine (N = 70), or fluoxetine, 20 mg q.d. (N = 33). Duloxetine dose was titrated in the first 3 weeks in a forced-titration regimen from 40 mg (20 mg b.i.d.) to 120 mg/day (60 mg b.i.d.). Patients were required to have a Clinical Global Impressions (CGI)-Severity of Illness scale score of at least moderate severity (> or = 4) and a 17-item Hamilton Rating Scale for Depression (HAM-D-17) total score of at least 15. Patients could not have had any current primary DSM IV Axis I diagnosis other than major depressive disorder, or any anxiety disorder as a primary diagnosis within the past year, excluding specific phobias. The primary efficacy measurement was the HAM-D-17 total score, and secondary measures included the Montgomery-Asberg Depression Rating Scale, CGI-Severity of Illness and CGI-Improvement, and Patient Global Impression of Improvement. Safety was evaluated by recording the occurrence of discontinuation rates and treatment emergent adverse events and by measurement of vital signs and laboratory analytes. RESULTS: Duloxetine was superior to placebo in change on the HAM-D-17 (p = .009). Estimated probabilities of response and remission were 64% and 56%, respectively, for duloxetine, compared with 52% and 30% for fluoxetine and 48% and 32% for placebo. Duloxetine was numerically superior to fluoxetine on the primary and most of the secondary outcome measures. In general, duloxetine was well tolerated; 76% of patients achieved the maximum dose, and insomnia and asthenia were the only adverse events reported statistically significantly (p < .05) more frequently by duloxetine-treated patients compared with placebo-treated patients. CONCLUSION: These data indicate that duloxetine is efficacious for the treatment of major depressive disorder and is well tolerated and safe. PMID- 11926723 TI - Switching patients from daily citalopram, paroxetine, or sertraline to once weekly fluoxetine in the maintenance of response for depression. AB - BACKGROUND: Major depressive disorder is frequently a chronic, recurrent condition necessitating maintenance treatment. For some patients, compliance with daily pharmacotherapy is difficult over time. As an alternative approach, a once weekly administered formulation of fluoxetine has recently been made available. This raises the important question of whether once-weekly enteric-coated fluoxetine, 90 mg, is effective for maintenance of response in patients whose depressive symptoms have responded to daily dosing with selective serotonin reuptake inhibitors (SSRIs) such as citalopram, paroxetine, or sertraline. METHOD: Patients had met DSM-IV criteria for major depressive disorder prior to beginning treatment for their current episode, had received 6 to 52 weeks of treatment with citalopram (20-40 mg/day [N = 83]), paroxetine (20 mg/day [N = 77]), or sertraline (50-100 mg/day [N = 86]), and had responded to that treatment (Clinical Global Impressions-Severity of Illness [CGI-S] score < or = 2, modified 17-item Hamilton Rating Scale for Depression [HAM-D-17] score < or = 10). Patients meeting these criteria (N = 246) continued treatment with their current SSRI for 1 week, then were switched to open-label enteric-coated fluoxetine, 90 mg, taken once weekly for 12 weeks. Safety measures were comparisons of spontaneously reported and solicited treatment-emergent adverse events. Efficacy measures were percentages of patients who discontinued the study for relapse and lack of efficacy and comparison of change from baseline to endpoint in scores on the modified HAM-D-17, subscales of the HAM-D-28, and the CGI-S. Quality of life measures were assessed with the MOS 36-Item Short-Form Health Survey (SF-36). We hypothesized that the once-weekly administration of fluoxetine could be safely and effectively initiated among subjects who had been stabilized on daily SSRI treatment. RESULTS: Seventy-nine percent of patients successfully completed a switch to enteric-coated fluoxetine, 90 mg, with 9.3% discontinuing due to relapse or lack of efficacy. Enteric-coated fluoxetine at a once-weekly dose of 90 mg was well tolerated in all groups. No significant increases were found in the HAM-D-17 total, HAM-D-28 subscores, or CGI-S score. Patients showed improvement from baseline to endpoint in most of the SF-36 health concepts. CONCLUSION: Enteric-coated fluoxetine taken once weekly appears to be well tolerated and efficacious in patients who responded to acute therapy with other SSRIs and were subsequently switched to fluoxetine once weekly for continuation/maintenance therapy. PMID- 11926724 TI - Treatment of borderline personality disorder with risperidone. AB - BACKGROUND: Of the various Axis II disorders, borderline personality disorder (BPD) is among the more critical to treat. There are at present few results in terms of clinical outcome with the psychotropic agents available. Possible targets for pharmacotherapy are affective symptoms, cognitive disturbances, and impulsive, self-injurious behaviors. In previous studies, atypical antipsychotics at low-to-moderate doses provided symptom reduction with good tolerability. Our purpose was to assess the efficacy of risperidone in BPD, focusing on its effects on impulsive-aggressive behavior. METHOD: Fifteen BPD outpatients (DSM-IV diagnosis) with prominent histories of aggressive behavior were included in an 8 week open-label study with risperidone at low-to-moderate doses. Axis II codiagnoses included antisocial personality disorder (N = 4). Exclusion criteria included current Axis I diagnosis or any major medical or neurologic illness. Efficacy measures were the 21-item Hamilton Rating Scale for Depression, the Brief Psychiatric Rating Scale, the DSM-IV Global Assessment of Functioning, and the self-rated Aggression Questionnaire. Evaluations were carried out at baseline and at the end of the treatment. RESULTS: Thirteen patients completed the trial; 2 patients dropped out because of lack of compliance. Final mean dose of risperidone was 3.27 mg/day. There was a significant (p = .0057) reduction in aggression based on Aggression Questionnaire scores. This amelioration was coupled with an overall improvement, including a reduction in depressive symptoms and an increase in energy and global functioning. CONCLUSION: Risperidone at low to-moderate doses can improve BPD symptomatology. Further studies are needed to explore the efficacy of risperidone versus placebo as well as in comparison to other potential treatments for BPD. PMID- 11926725 TI - Mechanisms of weight gain induced by antipsychotic drugs. PMID- 11926727 TI - Switch to mania after slow rTMS of the right prefrontal cortex. PMID- 11926726 TI - Delusions associated with quetiapine-related weight redistribution. PMID- 11926728 TI - Exacerbation of idiopathic priapism with risperidone-citalopram combination. PMID- 11926729 TI - Delirium caused by donepezil: a case study. PMID- 11926731 TI - Control of methicillin-resistant Staphylococcus aureus in hospitals. An impossible dream? PMID- 11926730 TI - Understanding changes in cholinergic function: implications for treating dementia. PMID- 11926733 TI - Interleukin-8 induction and adhesion of the coccoid form of Helicobacter pylori. AB - To determine the pathological significance of the coccoid form of Helicobacter pylori, its adhesion to and induction of secretion of interleukin-8 (IL-8) by gastric epithelial (MKN45) cells were studied. By flow cytometry, the adhesion of the coccoid form to MKN45 cells was significantly lower than that of the helical form. The monoclonal antibody A20 recognising lipopolysaccharide of H. pylori inhibited the adhesion of the coccoid form to MKN45 cells as much as that of the helical form. There was significantly lower induction of IL-8 secretion by MKN45 cells exposed to the coccoid form (two of four strains) as compared with the helical form. PMID- 11926732 TI - Characterisation of monoclonal antibodies against haemagglutinin associated with Clostridium botulinum type C neurotoxin. AB - Of 11 monoclonal antibodies (MAbs) prepared against the non-toxic component of type C Clostridium botulinum 16S toxin to clarify the function of the non-toxic component, seven recognised HA1, three recognised HA3b and one recognised HA2. Results of epitope mapping indicated that three of the seven anti-HA1 MAbs recognised the region between amino acid residues 121 and 140 and four recognised the three-dimensional structure of HA1. Three anti-HA3b MAbs recognised different regions between (approximately) amino acids 405-430, 180-270 and 275-297. The ability of these MAbs to interfere with binding of 16S toxin or non-toxic component, HA1 or HA3b to erythrocytes and to intestine tissue sections of guinea pig was observed. MAbs against HA3b and HA2 did not inhibit 16S toxin binding to either erythrocytes or epithelial cells, whereas some MAbs against HA1 did inhibit binding. The seven anti-HA1 MAbs can be classified into four groups based on their binding inhibition activities. The anti-HA1 MAbs that inhibited the binding of 16S toxin to the epithelial cells also neutralised or reduced the oral toxicity in mice, indicating that HA may play an important role in the absorption of the 16S toxin from the small intestine. PMID- 11926734 TI - Prevalence of serum antibodies to Helicobacter pylori VacA and CagA and gastric diseases in Chile. AB - The objective of this study was to evaluate the prevalence of antibodies to Helicobacter pylori CagA and VacA proteins and correlate this prevalence with gastric diseases in colonised Chileans. The study was performed in 418 adults colonised with H. pylori: 316 with gastroduodenal pathology (152 duodenal ulcer, 14 gastric cancer and 150 gastritis patients) and 102 asymptomatic subjects. Serum IgG antibodies to H. pylori were determined by enzyme immunoassay (EIA). Antibodies to VacA and CagA proteins were detected by Western blotting. In a subgroup of the patients, the vacuolating activity was determined by HeLa cell assay and the CagA product was confirmed by PCR assay. IgG antibodies to both VacA and CagA proteins of H. pylori were found in 270 (85%) of 316 colonised gastric patients and in 72 (71%) of 102 asymptomatic subjects. Colonisation with virulent strains was significantly higher among duodenal ulcer and gastric cancer patients than in gastritis patients or asymptomatic subjects. Infections with VacA+/ CagA+ H. pylori strains is common in Chile but, in contrast to some Asian countries, this phenotype was more prevalent in isolates from patients with more severe gastric pathologies. PMID- 11926735 TI - Gastric mucosal density of Helicobacter pylori estimated by real-time PCR compared with results of urea breath test and histological grading. AB - The accuracy of the urea breath test (UBT) and histological grading for estimation of the density of Helicobacter pylori in gastric mucosa is not known. Real-time (TaqMan) PCR was used to estimate the total number of H. pylori genomes in biopsy samples. These values were compared with those obtained by the UBT and the histological grade obtained by the Sydney system. The UBT and endoscopy with antral and corporal biopsies were performed in 88 consecutive untreated patients with dyspepsia. Bacterial culture and the rapid urease test were done with fresh biopsy materials. TaqMan PCR and histological examination were done on serial paraffin sections of the biopsy samples. Of the five methods tested, TaqMan PCR had the highest sensitivity and specificity (both 100%) in the diagnosis of H. pylori infection. The mean density of H. pylori genomes for pairs of biopsy samples from individual patients was compared with the individual values obtained by the UBT; correlation between the results was significant. The density of H. pylori genomes was higher in histological grades 1, 2 and 3 than in grade 0, without significant differences between adjacent grades from 1 to 3. These results suggest that the severity of H. pylori infection of the stomach can be estimated by the UBT and that histopathologists might state whether the organism is present or absent, rather than making a quantitative statement as recommended in the Sydney system. PMID- 11926736 TI - Prevalence of helicobacter pylori at oral and gastrointestinal sites in children: evidence for possible oral-to-oral transmission. AB - Acquisition of Helicobacter pylori occurs mainly in childhood. However, the mode of transmission remains unclear. To help elucidate this, 100 children attending for upper gastrointestinal endoscopy were investigated for the presence of H. pylori at various sites. H. pylori was detected in antral gastric biopsies by the rapid urease test (13 patients), culture (13 patients), histology (15 patients) and PCR (20 patients). Gastric juice was positive for H. pylori in 3 patients by culture and 11 patients by PCR. The dental plaque from 68% of gastric biopsy positive patients (as determined by culture or PCR) and 24% of gastric biopsy negative patients was positive for H. pylori by PCR. The presence of H. pylori in dental plaque was significantly associated with the presence of this organism in the stomach. H. pylori was detected by PCR in the faeces of 25% of gastric biopsy positive children sampled. H. pylori was not cultured on any occasion from the oral cavity or faeces. The evidence from this study suggests that oral-to-oral transmission may be a possible mode of spread of H. pylori in children. PMID- 11926738 TI - Antibodies to granulocytic ehrlichiae in cattle from Connecticut. AB - Enzyme-linked immunosorbent assays (ELISA) with a purified recombinant 44-kDa protein and indirect fluorescent antibody (IFA) staining methods incorporating whole-cell antigens of the human granulocytic ehrlichiosis (HGE) agent were used to detect antibodies to Ehrlichia phagocytophila genogroup organisms in cattle sera. The cattle lived in tick-infested areas of Connecticut, USA and were healthy at the times blood samples were collected in 1990, 1999 and 2000. Of the 339 serum samples analysed, 40 (12%) and 15 (4%) were positive by ELISA and IFA, respectively. Western immunoblots of a subset of sera verified antibody reactivity of six serum samples, positive by ELISA with titres of 640-2,560, to a protein with a molecular mass of c. 44 kDa. Although seroprevalence rates were low, cattle were exposed to the HGE agent at different sites and should be monitored for anaemia, leukopenia or thrombocytopenia, especially if there is evidence of unexplained decreased milk production. Different serological testing methods should be used to detect immunoglobulins. PMID- 11926737 TI - Genetic analysis of the outer surface protein C gene of Lyme disease spirochaetes (Borrelia burgdorferi sensu lato) isolated from rodents in Taiwan. AB - The outer surface protein C gene (ospC) of Lyme disease spirochaetes (Borrelia burgdorferi sensu lato) was analysed for the first time in Taiwan. The genetic identities of these Taiwan isolates (TWKM1-7) were determined by restriction fragment length polymorphism (RFLP) analysis and sequence similarities of the PCR amplified ospC gene amplicons. After cleavage by nuclease Dral, differential fragment patterns of PCR-amplified ospC DNA in relation to different genospecies of Lyme disease spirochaetes were observed and all of these Taiwan isolates were genetically affiliated to the genospecies of B. burgdorferi sensu stricto. The phylogenetic analysis on the sequence similarity of these Taiwan isolates revealed a highly homogeneous genotype, ranging from 99.3% to 100%, within the genospecies of B. burgdorferi sensu stricto and was distinguished from other genospecies of Borrelia isolates. The sequence similarity analysis also revealed the high sequence variability of the ospC gene among Borrelia strains that belong to the same genospecies but were isolated from different biological and geographical sources. Thus, these results provide the first investigation on the genetic identity of the ospC gene of these Taiwan isolates and show that these Taiwan isolates were closely related genetically to the genospecies of B. burgdorferi sensu stricto. PMID- 11926739 TI - Isolation and characterisation of Shiga toxigenic Escherichia coli strains from northern Palestine. AB - Shiga toxigenic Escherichia coli (STEC) isolates from symptomatic and asymptomatic patients in northern Palestine in 1999 were screened for serotype O157 and characterised for virulence genes by multiplex PCR assay. Of the 176 STEC isolates, 124 (70.5%) were of serotype O157. All these isolates carried the gene for Shiga toxin type 1 (stx,) and 112 (90.3%) carried stx2. The intimin encoding gene locus eae was detected in 16 isolates (12.9%) and the enterohaemolysin encoding gene, hlyA, in 18 (14.5%). Statistical analysis showed a significant association between the presence of eaeA and hlyA, either alone or combined with stx1 and stx2 genes in O157 isolates from symptomatic infection. ERIC-PCR analysis of DNA from 80 serotype O157 isolates revealed three major clonal populations. PMID- 11926741 TI - Mixed species biofilms of Candida albicans and Staphylococcus epidermidis. AB - A simple catheter disk model system was used to study the development in vitro of mixed species biofilms of Candida albicans and Staphylococcus epidermidis, two organisms commonly found in catheter-associated infections. Two strains of S. epidermidis were used: a slime-producing wild type (strain RP62A) and a slime negative mutant (strain M7). In mixed fungal-bacterial biofilms, both staphylococcal strains showed extensive interactions with C. albicans. The susceptibility of 48-h biofilms to fluconazole, vancomycin and mixtures of the drugs was determined colorimetrically. The results indicated that the extracellular polymer produced by S. epidermidis RP62A could inhibit fluconazole penetration in mixed fungal-bacterial biofilms. Conversely, the presence of C. albicans in a biofilm appeared to protect the slime-negative staphylococcus against vancomycin. Overall, the findings suggest that fungal cells can modulate the action of antibiotics, and that bacteria can affect antifungal activity in mixed fungal-bacterial biofilms. PMID- 11926740 TI - Experimental infection of germ-free mice with hyper-toxigenic enterohaemorrhagic Escherichia coli O157:H7, strain 6. AB - A mouse enterohaemorrhagic Escherichia coli (EHEC) infection model was developed with a combination of germ-free (GF) mice and hyper-toxigenic EHEC (HT-EHEC) O157:H7 strain 6. The HT-EHEC strain 6 produced both Shiga-like toxin (SLT)-1 1.0 microg/mI and SLT-2 8.2 microg/ml in vitro. Eight-week-old germ-free mice were inoculated intragastrically with 5.0 x 10(7) cfu of HT-EHEC strain 6. All GF mice challenged with the HT-EHEC developed ruffled fur and convulsion of limbs or hindleg weakness within 3 days after the challenge, culminating in death within 5 days. The HT-EHEC colonised well at a level of 10(8) - 10(9) cfu/g of faeces 5 days after the challenge. Both SLT-1 and SLT-2 were demonstrated in the faeces of the mice for 5 days after challenge. Histological examination of the colons of the infected mice showed congestion of the lamina propria mucosa, mild inflammatory cell infiltration and goblet cell depletion. In proximal tubules of the renal cortex, epithelial swelling with scattered necrotic cells was recognised. Endothelial swelling and mononuclear cell infiltration were also observed in the glomeruli. The brain showed acute neuronal necrosis in the cerebrum and slight loss of Purkinje cells in the cerebellum. Passive immunisation with anti-SLT antisera prolonged the life of the mice without anyneural symptoms. Microscopically, all tissue specimens from the passively immunised mice were not remarkable. These results indicate that the infection of GF mice with HT-EHEC is a useful animal model to study the pathogenicity of SLT producing E. coliand the toxins. PMID- 11926742 TI - Protective immunity against lethal HSV-1 challenge in mice by nucleic acid-based immunisation with herpes simplex virus type-1 genes specifying glycoproteins gB and gD. AB - DNA-based vaccines were employed to assess protective immunity against herpes simplex virus in experimental infections of hairless (strain SKH1) and BALB/c mice. Mice were vaccinated with plasmids containing the herpes simplex virus type 1 (HSV-1) glycoprotein B (gB) or D (gD) genes under the human cytomegalovirus immediate-early promoter control. Vaccines were injected intramuscularly (i.m.) or intraperitoneally (i.p.) as purified DNA alone or as formulations supplemented with different non-ionic block copolymers. Antibody responses were assessed by immunofluorescence and radio-immunoprecipitation assays. Mice inoculated with either gB or gD plasmid, alone or with non-ionic block copolymers CRL 1029 and CRL 1190, produced high levels of antibodies specific for gB or gD. Three weeks after the last vaccination, mice were challenged with a clinical HSV-1 isolate (ABGK-1) by inoculation of a shaved and subsequently scarified area between the third and fourth lumbar vertebrae. Mice immunised with either gD or gB plasmid alone or mixed with copolymers were protected against lethal HSV-1 challenge when immunisation was performed via the i.m. route. Immunisations given via the i.p. route induced humoral responses in some mice and protected the animals against lethal HSV-1 challenge only when the formulations contained copolymers. The BALB/c mouse model was shown to be as good a model as the hairless mouse model. PMID- 11926743 TI - Cytomegalovirus in solid organ transplantation. AB - Despite substantial advances made in controlling the effects of cytomegalovirus (CMV) infection, it remains the single most important pathogen in solid organ transplantation (SOT). Because CMV shares some characteristics with other human herpesviruses, it is also an important model system for understanding the actions of herpesviruses 6 and 7, Epstein-Barr virus (EBV) and, potentially, hepatitis C and B. As the lessons learned from HIV influenced our thinking about other viral infections (e.g. importance of viral load), so too what is learned about CMV will be applied to other herpesviruses. The pervasive nature of CMV and the common problems posed by this virus prompted the convening of a panel of experts in the field of SOT to discuss issues associated with CMV in transplant recipients. This supplement reflects the presentations and discussions at this symposium, including the clinical implications of CMV drug resistance, economic impact of CMV on transplant programs, the rationale for CMV hyperimmune globulin (CMW-IGIV, CytoGam) in SOT, antibody inhibition of CMV, hypogammaglobulinemia, role of CMV in allograft vasculopathy, and the clinical use of CytoGam therapy in a variety of SOT patients. A number of questions during the general discussion prompted the addition of other material to this Supplement, including the development of CMV IGIV for clinical use in SOT recipients and resource utilization associated with CMV-related hospital readmissions. PMID- 11926744 TI - Economic impact of cytomegalovirus in solid organ transplantation. AB - Cytomegalovirus (CMV) infection has a direct effect on morbidity in solid organ transplantation patients, and indirect effects related to the development of opportunistic infections, allograft rejection, and patient mortality. Although intuitively it follows that costs attributable to CMV infections would be increased, direct proof has remained elusive. Accumulating evidence suggests, however, that CMV infection has a significant impact on the costs to transplantation programs, particularly in seronegative recipients of seropositive allografts (D+/R-), and additional costs may be incurred through the effects on CMV potentiating the risks of various opportunistic infections leading to graft rejection. PMID- 11926745 TI - Cytomegalovirus drug resistance and clinical implications. AB - Antiviral agents are commonly used for cytomegalovirus (CMV) prophylaxis or therapy after solid organ transplantation. Until recently, the detection of drug resistant CMV in this setting was rare, but ganciclovir resistance has now been reported to occur in 5-10% of high-risk patient subsets, such as those undergoing primary CMV infection. Persistent viral shedding or progressive CMV disease after several weeks of antiviral therapy may indicate a problem with drug resistance, though laboratory testing is required to confirm this. Rapid genotypic assays for specific mutations in the viral UL97 phosphotransferase or UL54 DNA polymerase genes can be used to detect resistance and predict cross-resistance to other drugs. The emergence of drug resistance may be reduced by optimization of host immunity, use of potent antiviral drug regimens, and adherence to dosing regimens that adequately suppress viral replication. PMID- 11926746 TI - Human CMV-IGIV (CytoGam) neutralizes human cytomegalovirus (HCMV) infectivity and prevents intracellular signal transduction after HCMV exposure. AB - Pretreatment of human cytomegalovirus (HCMV) with human hyperimmune globulin (CytoGam) in human embryonic lung (HEL) fibroblast culture showed successful inhibition of infectivity, and decreased extracellular viral titers and extracellular viral DNA. CytoGam prevented HCMV from inducing intracellular activation of NF-kappaB, Sp-1, and P13-K signaling pathways and the production of immediate-early (IE), early (E), and late (L) viral proteins. CytoGam neutralization of HCMV in this cell culture model prevented the earliest known signal transduction events (NF-kappaB, Sp-1, P13-K activation) after viral specific glycoproteins bind to their cognate cell membrane receptors, suggesting that this agent contains highly effective neutralizing antibodies against HCMV. PMID- 11926747 TI - Antibody inhibition of cytomegalovirus: the role of natural killer and macrophage effector cells. AB - To explore mechanisms by which antibody might inhibit cytomegalovirus (CMV), we measured the ability of intravenous CMV-IgG (CytoGam) to reduce viral yield in the presence of effector cells. Foreskin fibroblasts were infected with a clinical strain of CMV, and CytoGam was added along with effector cells consisting of either unfractionated peripheral blood mononuclear cells (PBMCs), natural killer (NK) cells, or macrophages. The combination of CytoGam and either of the effector cell types markedly inhibited established CMV infection in vitro. In addition, CytoGam combined with effector cells protected the monolayer from CMV-induced cytopathic effects. Antibody-dependent, effector cell-mediated functions may underlie the ability of CytoGam to prevent or modulate CMV infection in vivo. PMID- 11926749 TI - Hypogammaglobulinemia after heart transplantation: impact of pre-emptive use of immunoglobulin replacement (CytoGam) on infection and rejection outcomes. AB - Hypogammaglobulinemia (HGG) in solid organ transplant (SOT) patients confers an increased risk of opportunistic infections and poorer outcomes. Severe HGG (IgG < 350 mg/dL) after heart transplantation may follow intensification of immunosuppressive therapy and the resultant increased risk of opportunistic infections, particularly cytomegalovirus (CMV) disease. Evaluation of the effects of replacement therapy using intravenous immunoglobulin (CMV-IGIV, CytoGam) was conducted in cardiac transplant recipients and the data matched with a historical control group. Patients with severe HGG who received pre-emptive replacement therapy had significantly fewer opportunistic infections (P < 0.001) and episodes of rejection (grade > or = 3; P = 0.03 and grade > or = 2; P = 0.04) compared with the control group. PMID- 11926748 TI - Cytomegalovirus prophylaxis with ganciclovir and cytomegalovirus immune globulin in liver and intestinal transplantation. AB - Liver and intestinal transplant recipients at the University of Miami receive an intensive regimen of cytomegalovirus (CMV) prophylactic therapy consisting of a combination of CMV immune globulin intravenous (CMV-IGIV, CytoGam) and ganciclovir. The 5-year experience with this regimen in liver transplant patients showed effective CMV prophylaxis in this patient population. The importance of an effective prophylactic strategy was underscored by higher observed rates of chronic rejection and post-transplant lymphoproliferative disorder (PTLD) in CMV infected patients. The use of CMV-positive donors for intestinal transplants did not increase the incidence of CMV disease. Intestinal transplant recipients had improved survival rates, reflecting an aggressive policy of monitoring, immunosuppression, and CytoGam plus ganciclovir prophylaxis. PMID- 11926750 TI - Role of cytomegalovirus in cardiac allograft vasculopathy. AB - Cardiac allograft vasculopathy is the most common cause of death and retransplantation following heart transplantation, and about 10% of patients per year have evidence of accelerated vascular disease; 50% at 5 years. Cytomegalovirus (CMV) infection has been associated with accelerated cardiac vasculopathy and decreased 5-year survival. Prophylactic therapy using ganciclovir has reduced the incidence of CMV disease, but not in the group at highest risk, namely the seronegative recipient of an allograft from a seropositive donor (D+/R-). Combination prophylaxis consisting of CMV hyperimmune globulin (CMV-IGIV) plus ganciclovir is associated with decreased intimal thickening, reduced coronary artery disease and obliterative bronchiolitis, and improved survival. PMID- 11926751 TI - Use of cytomegalovirus immune globulin and ganciclovir for the prevention of cytomegalovirus disease in lung transplantation. AB - Cytomegalovirus (CMV) infection and disease continue to be significant causes of morbidity and mortality in lung transplant recipients. The potential benefits of CMV prophylaxis extend beyond prevention of the immediate CMV infection to potentially preventing CMV-associated complications, including superinfection due to Aspergillus bacteria, and other opportunistic infections, and bronchiolitis obliterans syndrome (BOS). Longer courses of prophylactic intravenous (IV) ganciclovir, sequential IV/oral therapy, addition of intravenous CMV immune globulin (CMV-IGIV), surveillance tests, and investigation of the role of hypogammaglobulinemia are a few of the strategies and issues being evaluated to improve CMV prophylaxis and, consequently, graft and patient survival. PMID- 11926752 TI - Multicenter US study of hospital resource utilization associated with cytomegalovirus-related readmission of renal and heart transplant patients. AB - Incidence of cytomegalovirus (CMV)-related rehospitalization and associated resource use were captured by the Transplant Infection Cost Analysis (TICA) program, which examined patient records and hospital billing data in multiple solid organ transplant centers in the US. The experiences of two adult heart and three adult renal transplant centers were each pooled for analysis. Financial data were standardized to 1998 US dollars using the Medical Care component of the US Consumer Price Index. CMV-related readmissions among renal transplant patients averaged 10.5 days (range 1-56) with average charges of $22,598. Heart transplant patients readmitted for CMV incurred an average charge of $42,111 and average hospital stay of 10.9 days (range 2-95). CMV-related hospital resource use represented a significant portion of the average cost of the original transplant and associated length of stay. PMID- 11926753 TI - Historical overview of the use of cytomegalovirus hyperimmune globulin in organ transplantation. AB - A historical review of the development of cytomegalovirus hyperimmune globulin (CMV-IG, CytoGam) shows its increasing use in solid organ transplant (SOT) recipients, either alone or in combination with ganciclovir. A review of clinical trials of CytoGam in renal transplant recipients shows reductions in CMV associated syndromes and fungal and parasitic superinfections, and increases in graft survival, while CytoGam prophylaxis trials in orthotopic liver transplant (OLT) recipients have produced reductions in severe CMV-associated disease and invasive fungal disease. A combination of CytoGam plus ganciclovir in OLT recipients has resulted in reductions in CMV hepatitis and infection, and CMV disease and viremia, plus a trend in improved 1- and 2-year survival rates. PMID- 11926754 TI - The preclinical history of high-frequency ventilation. AB - As with many innovative techniques in modern medicine, high-frequency ventilation (HFV) was discovered by accident rather than by design. Various modes of HFV also were developed concurrently, with little or no attempt to standardize nomenclature. High-frequency ventilation developed along three main tracks, driven by somewhat different forces. PMID- 11926755 TI - The history of high-frequency ventilation. AB - High-frequency ventilation was first introduced 30 years ago as a method for reducing intrathoracic pressure during thoracic and laryngeal surgery. High frequency oscillation was developed in the 1970's for the treatment of lung disease of prematurity but is now used for acute hypoxemic respiratory failure in all ages. High-frequency jet ventilation is still most commonly used as a rescue therapy. PMID- 11926757 TI - High-frequency ventilation design and equipment issues. AB - Various technical approaches to high-frequency ventilation are available. Of these, HFOV is used most commonly. As high-frequency ventilation becomes increasingly used in the care of adult patients, the technical issues related to this therapy will become increasingly important. PMID- 11926756 TI - Overview of high-frequency ventilation modes, clinical rationale, and gas transport mechanisms. AB - HFV has been demonstrated to be a safe and effective way to ventilate and oxygenate patients, both short- and long-term, when used by experienced practitioners. It has carved out a niche in the specific management of respiratory problems in children and neonates; however, as understanding of the variables that independently contribute to VILI evolves, it is becoming clear that this mode of ventilation may be suited to the goals of lung protection. In addition, it is accepted also that initial assessment of HFV as a lung-protective strategy has failed to take into consideration significant variables that have been shown to be important in animal studies. This may have caused this mode of ventilation to be over looked as a possible strategy in the management of patients with severe lung disease. A number of trials are underway using ventilatory approaches based on current concepts of VILI, including improved CMV strategies. It is hoped that the results of these studies will identify the future role for HFV in the clinical setting. PMID- 11926758 TI - High-frequency jet ventilation. AB - High-frequency ventilation, including HFJV, is an interesting alternative approach to mechanical ventilatory support that may offer benefits in terms of improved gas exchange and lower maximal alveolar distending pressures. Clinical data demonstrating improved outcome exist for neonatal and some forms of pediatric respiratory failure. No such data, however, exist for adults. Important complications can develop, and an extensive learning curve is required for operators to become skilled at delivering proper support safely. Presently, HFV should be limited to only specific applications (e.g., selected neonates, adult airway surgical procedures) and to centers skilled in its use. Considerably more data are required before extensive application, especially in the adult, is warranted. PMID- 11926759 TI - The science of neonatal high-frequency ventilation. AB - Despite improvements in respiratory care, ventilator-induced lung injury remains an important cause of morbidity and mortality in neonates who require assisted ventilation. Animal data clearly demonstrate that high-frequency ventilation can be used successfully to reduce lung injury in experimental models of acute lung injury. These models and human research show that the efficacy of high-frequency ventilation is dependent on optimizing functional residual capacity and avoiding lung overinflation. When used with a strategy that promotes lung recruitment, high-frequency ventilation effectively reduces the occurrence of chronic lung disease and is not associated with significant brain injury. When used with a strategy that allows the lung to collapse or is associated with hyperventilation, however, high-frequency ventilation does not reduce lung injury and is associated with significant brain injury. Like every tool we use to support critically ill neonates, high-frequency ventilation needs a careful carpenter. As therapies and health care strategies evolve, there remains nothing more important than the health care team at the bedside. Critical evaluation of the patient and his or her response to the therapy being offered is essential to promotion of the patient health outcome. PMID- 11926760 TI - High-frequency oscillatory ventilation in pediatric patients. AB - HFOV is a mode of ventilation that can achieve oxygenation and ventilation while maintaining maximal lung recruitment on the deflation limb of its pressure-volume curve. The primary theoretical advantages of HFOV over CMV in the management of acute lung injury are that HFOV allows adequate alveolar ventilation with minimal peak-trough pressure changes, provides lung recruitment, and avoids end inspiratory overdistension of the relatively compliant nondependent lung. Taken together, the results of studies in animals, preterm and term neonates, and older pediatric patients reveal that an "open-lung" strategy, with the goal of a high end-expiratory lung volume, is safe and superior to CMV in both the short-term (rapidly improved oxygenation and/or ventilation) and longer-term (lower incidence of chronic lung disease). The improved longer-term clinical outcomes on HFOV are presumably because of less ventilator-induced lung injury. As experience with HFOV in older patients grows, ventilator technology matures, and understanding of the pathophysiology of acute respiratory distress syndrome (RDS) deepens, it is likely that HFOV will find widespread use for the management of respiratory failure caused by acute lung injury in patients from preterm neonates to adults. PMID- 11926761 TI - The use of high-frequency oscillatory ventilation in adults with acute lung injury. AB - The use of HFOV in adults is still in its infancy. There is, however, much promise to support further study of this ventilatory modality. Rescue case series have shown that HFOV is effective in improving gas exchange and appears safe in this group of extremely ill patients. In addition, as evidence continues to mount regarding the importance of VILI and its mechanisms, HFOV provides a theoretically attractive alternative to conventional lung-protective ventilatory modes. When HFOV is used in adults, it should be in conjunction with an effort to recruit atelectatic lung units by employing higher mean airway pressures and weaning FIO2 before P(AW). HFOV could be used as one of a number of new therapies for the patient failing to oxygenate on CMV. Its routine use to prevent VILI cannot be recommended at this time, as no data are available. Further clinical studies potentially leading to a large randomized controlled trial of HFOV versus best conventional therapy appear worth pursuing. PMID- 11926762 TI - Combination therapy. AB - Although not allowing one to state conclusively that a particular combination of approaches to CMV and adjuncts to CMV affect outcome, the results are encouraging. Independently, none of the modalities discussed-HFV, ECMO, NO, or PLV-have shown a statistically significant improvement in outcome except NO. With NO, improvement is only in a small select group of patients; however, in combination there may be greater overall impact on outcome. With each of these modalities, more information is needed on how to apply each technique optimally. It is easy to see that modification of the experimental protocol in each study may have altered results. Clearly, more laboratory and clinical studies examining the combined use of these therapies are needed, as is identification of the specific patient populations where these combined therapies may be most effective. PMID- 11926763 TI - Implementing and troubleshooting high-frequency oscillatory ventilation in adults in the intensive care unit. AB - This article provides general guidelines for application and titration of HFOV; however, the use of HFOV in adults with ARDS still is considered experimental. Further evaluation of the efficacy of HFOV and potential impact on outcomes are needed in adults, particularly in the form of randomized, controlled trials. PMID- 11926764 TI - Inflammation in acute coronary syndromes. PMID- 11926765 TI - Carotid lesions as risk factor for ischemic heart disease. PMID- 11926766 TI - Peripheral arterial disease as global vascular risk factor. PMID- 11926767 TI - Efficacy and safety of aspirin in the long-term management of atherothrombosis. PMID- 11926768 TI - Antioxidant therapy in vascular disease. PMID- 11926769 TI - The long-term use of blockers of the platelet ADP receptor in acute coronary syndromes. PMID- 11926770 TI - Genetic polymorphisms of clotting factors and coronary heart disease. PMID- 11926771 TI - Bioactive lipids and atherothrombosis. PMID- 11926772 TI - Platelets and oxygen radicals: mechanisms of functional modulation. PMID- 11926773 TI - Inhibition of cholesterol biosynthesis as a new antithrombotic strategy. PMID- 11926774 TI - Antiaggregant therapy and/or anticoagulant therapy in the cerebrovascular patient. PMID- 11926775 TI - Clopidogrel and the CURE results. PMID- 11926776 TI - Thrombolytic therapy, angioplasty or something else: which is the therapeutic paradigm in patients with acute myocardial infarction? PMID- 11926777 TI - Glycoprotein IIb/IIIa antagonists in acute coronary syndromes. PMID- 11926778 TI - Home treatment and secondary prevention of deep vein thrombosis. PMID- 11926780 TI - New pharmacologic strategies for arterial and venous thromboembolism. PMID- 11926779 TI - Heparins: their established role in acute coronary syndromes and perspectives in atrial fibrillation. PMID- 11926781 TI - Overview of the clinical results of pentasaccharide in major orthopedic surgery. PMID- 11926782 TI - New antithrombotic agents in the management of venous thromboembolism. PMID- 11926783 TI - Hypercoagulability and atherothrombosis: clinical suggestions and perspectives. PMID- 11926784 TI - Effect of aerobic exercise on blood pressure: a meta-analysis of randomized, controlled trials. AB - PURPOSE: Physical activity has been associated with reduced blood pressure in observational epidemiologic studies and individual clinical trials. This meta analysis of randomized, controlled trials was conducted to determine the effect of aerobic exercise on blood pressure. DATA SOURCES: English-language articles published before September 2001. STUDY SELECTION: 54 randomized, controlled trials (2419 participants) whose intervention and control groups differed only in aerobic exercise. DATA EXTRACTION: Using a standardized protocol and data extraction form, three of the investigators independently abstracted data on study design, sample size, participant characteristics, type of intervention, follow-up duration, and treatment outcomes. DATA SYNTHESIS: In a random-effects model, data from each trial were pooled and weighted by the inverse of the total variance. Aerobic exercise was associated with a significant reduction in mean systolic and diastolic blood pressure (-3.84 mm Hg [95% CI, -4.97 to -2.72 mm Hg] and -2.58 mm Hg [CI, -3.35 to -1.81 mm Hg], respectively). A reduction in blood pressure was associated with aerobic exercise in hypertensive participants and normotensive participants and in overweight participants and normal-weight participants. CONCLUSIONS: Aerobic exercise reduces blood pressure in both hypertensive and normotensive persons. An increase in aerobic physical activity should be considered an important component of lifestyle modification for prevention and treatment of high blood pressure. PMID- 11926785 TI - Administration of neutral protamine Hagedorn insulin at bedtime versus with dinner in type 1 diabetes mellitus to avoid nocturnal hypoglycemia and improve control. A randomized, controlled trial. AB - BACKGROUND: Intensive insulin treatment of type 1 diabetes mellitus increases the risk for nocturnal hypoglycemia. OBJECTIVE: To demonstrate that splitting the evening insulin regimen reduces the risk for nocturnal hypoglycemia in intensive treatment of type 1 diabetes mellitus. DESIGN: Randomized, open, two-treatment crossover trial in two 4-month periods. SETTING: University research center in Italy. PATIENTS: 22 C-peptide-negative persons with type 1 diabetes mellitus (mean age [+/-SD], 29 +/- 3 years). INTERVENTIONS: Each patient was randomly assigned to one of two insulin regimens for 4 months and then switched to the other regimen for another 4 months. The two treatment regimens were 1) mixed treatment--a mixture of human regular and neutral protamine Hagedorn (NPH) insulin administered before dinner and 2) split treatment--human regular insulin administered at dinner and NPH insulin administered at bedtime. MEASUREMENTS: Frequency of nocturnal hypoglycemia. Secondary end points were levels of fasting blood glucose and hemoglobin A1c and responses to experimental hypoglycemia. RESULTS: During the split-regimen treatment period, patients had fewer episodes of nocturnal hypoglycemia (mean [+/-SE], 0.10 +/- 0.02 episode/patient-day vs. 0.28 +/- 0.04 episode/patient-day; P = 0.002), a lower fasting blood glucose level (mean [+/-SE], 7.6 +/- 0.2 mmol/L vs. 8.3 +/- 0.5 mmol/L [137 +/- 4 mg/dL vs. 160 +/- 8 mg/dL]; P = 0.030), less variable fasting blood glucose levels (SD range, 2.0 +/- 0.4 vs. 3.5 +/- 0.6; P = 0.001), and lower hemoglobin A1c value (mean [+/-SE], 7.0% +/- 0.11% vs. 7.5% +/- 0.15%; P = 0.004) than during the mixed regimen. Responses to experimental hypoglycemia were better preserved with the split regimen than with the mixed regimen. CONCLUSION: When the goal of insulin therapy in type 1 diabetes mellitus is near-normoglycemia, splitting the evening insulin treatment regimen into short-acting insulin at dinner and NPH insulin at bedtime reduces the risks for nocturnal hypoglycemia and hypoglycemia unawareness and decreases the hemoglobin A1c value compared with mixing short acting insulin and NPH insulin at dinner. PMID- 11926786 TI - Combination therapy with oral sildenafil and inhaled iloprost for severe pulmonary hypertension. AB - BACKGROUND: Inhalation of the stable prostacyclin analogue iloprost is being studied for treatment of pulmonary hypertension. The selective phosphodiesterase 5 inhibitor sildenafil has been reported to cause pulmonary vasodilatation. OBJECTIVE: To evaluate the safety and effectiveness of oral sildenafil, alone and in combination with inhaled iloprost, for treatment of pulmonary hypertension. DESIGN: Randomized, controlled, open-label trial. SETTING: Intensive care unit. PATIENTS: 30 patients with severe pulmonary arterial hypertension (n = 16), chronic thromboembolic pulmonary hypertension (n = 13), or pulmonary hypertension due to aplasia of the left pulmonary artery (n = 1), all classified as New York Heart Association class III or IV. INTERVENTION: All patients received inhaled nitric oxide and aerosolized iloprost (inhaled dose, 2.8 microg). They were then randomly assigned to receive 12.5 mg of oral sildenafil, 50 mg of sildenafil, 12.5 mg of sildenafil plus inhaled iloprost, or 50 mg of sildenafil plus inhaled iloprost. MEASUREMENTS: Systemic and pulmonary arterial pressure, pulmonary arterial occlusion pressure, cardiac output, central venous pressure, peripheral arterial oxygen saturation, and arterial and mixed venous blood gases were measured during right-heart catheterization by using a Swan-Ganz catheter. RESULTS: In rank order of pulmonary vasodilatory potency (maximum reduction of pulmonary vascular resistance and increase in cardiac index), 50 mg of sildenafil plus iloprost was most effective, followed by 12.5 mg of sildenafil plus iloprost. Iloprost alone and 50 mg of sildenafil were almost equally effective but were less potent than the combination regimens, and the least potent treatments were 12.5 mg of sildenafil and nitric oxide. In patients who received 50 mg of sildenafil plus iloprost, the maximum change in pulmonary vasodilatory potency was -44.2% (95% CI, -49.5% to -38.8%), compared with -14.1% (CI, -19.1% to -9.2%) in response to nitric oxide. With administration of 50 mg of sildenafil plus iloprost, the area under the curve for reduction in pulmonary vasodilatory resistance surpassed that of administration of 50 mg of sildenafil alone and iloprost alone combined, the vasodilatory effect lasted longer than 3 hours, and systemic arterial pressure and arterial oxygenation were maintained. No serious adverse events occurred. CONCLUSION: Although limited by the small sample and lack of long-term observations, the study shows that oral sildenafil is a potent pulmonary vasodilator that acts synergistically with inhaled iloprost to cause strong pulmonary vasodilatation in both severe pulmonary arterial hypertension and chronic thromboembolic pulmonary hypertension. PMID- 11926787 TI - Change in coronary flow reserve on transthoracic Doppler echocardiography after a single high-fat meal in young healthy men. AB - BACKGROUND: High-fat meals and elevated triglyceride levels are associated with cardiovascular disease. In recent studies of brachial artery vasoactivity, a single high-fat meal reduced endothelial function in young healthy men. It is unknown whether coronary microcirculation is affected after high-fat meals. OBJECTIVE: To evaluate change in coronary flow reserve after a single high-fat meal. DESIGN: Controlled interventional study. SETTING: University hospitals. PATIENTS: 15 young healthy men (mean age [+/-SD], 29 +/- 4 years). INTERVENTION: Coronary flow reserve was determined by using transthoracic Doppler echocardiography before and after consumption of a high-fat meal in all 15 men and before and after consumption of a low-fat meal in 5 of 15 men. MEASUREMENTS: Coronary flow reserve, lipid levels, and hemodynamic characteristics. RESULTS: In all men, triglyceride levels increased significantly from baseline 5 hours after the high-fat meal, from 1.1 mmol/L to 2.8 mmol/L (100 mg/dL to 250 mg/dL) (P < 0.001). Average coronary flow reserve was 4.02 before and 3.30 5 hours after the high-fat meal (decrease, 18% [95% CI, 13% to 23%]). In the 5 men who received both meals, mean coronary flow reserve decreased by 0.79 after the high-fat meal and increased by 0.07 after the low-fat meal (difference, -0.86 [CI, -1.36 to 0.37]; P = 0.03). Mean triglyceride levels increased by 1.6 mmol/L (140 mg/dL) after the high-fat meal and 0.1 mmol/L (10 mg/dL) after the low-fat meal (difference, 1.5 mmol/L [CI, 0.3 to 2.7 mmol/L], 130 mg/dL [CI, 23 to 236 mg/dL]; P = 0.03). CONCLUSIONS: Coronary flow reserve decreased after a single high-fat meal in young healthy men. High-fat meals may be detrimental to coronary microcirculation. PMID- 11926788 TI - Is C-reactive protein specific for vascular disease in women? AB - BACKGROUND: C-reactive protein (CRP) predicts risk for future cardiovascular events in asymptomatic individuals. However, because CRP also predicts total mortality, its specificity for vascular disease is uncertain. OBJECTIVE: To compare the predictive value of CRP for cancer and cardiovascular disease, the major determinants of mortality. DESIGN: Prospective, nested case-control study. SETTING: The Women's Health Study, an ongoing prospective cohort study involving 28345 U.S. women 45 years of age and older who were healthy at the time of enrollment. PARTICIPANTS: 643 women who subsequently developed cancer or had cardiovascular events; 643 age- and smoking-matched women who remained free of either disease during 58-month follow-up. MEASUREMENTS: Baseline CRP levels. RESULTS: Little evidence showed that increasing quartiles of baseline CRP predicted incident cancer (adjusted relative risks, 1.0, 1.2, 1.1, and 1.3; P for trend > 0.2). In contrast, increasing quartiles of baseline CRP were a strong marker of risk for future cardiovascular disease (adjusted relative risks, 1.0, 2.9, 3.4, and 5.6; P for trend < 0.001). CONCLUSION: C-reactive protein appears to independently predict cardiovascular events but not cancer. PMID- 11926789 TI - Acquired aplastic anemia. AB - In aplastic anemia, hematopoiesis fails: Blood cell counts are extremely low, and the bone marrow appears empty. The pathophysiology of aplastic anemia is now believed to be immune-mediated, with active destruction of blood-forming cells by lymphocytes. The aberrant immune response may be triggered by environmental exposures, such as to chemicals and drugs or viral infections and, perhaps, endogenous antigens generated by genetically altered bone marrow cells. In patients with post-hepatitis aplastic anemia, antibodies to the known hepatitis viruses are absent; the unknown infectious agent may be more common in developing countries, where aplastic anemia occurs more frequently than it does in the West. The syndrome paroxysmal nocturnal hemoglobinuria (PNH) is intimately related to aplastic anemia because many patients with bone marrow failure have an increased population of abnormal cells. In PNH, an entire class of proteins is not displayed on the cell surface because of an acquired X-chromosome gene mutation. The PNH cells may have a selective advantage in resisting immune attack. In contrast, the disease myelodysplasia can be confused with aplasia and can also evolve from aplastic anemia. The occurrence of cytogenetic abnormalities in patients years after presentation implies that genomic instability is a feature of this immune-mediated disease. Aplastic anemia can be effectively treated by stem-cell transplantation or immunosuppressive therapy. Transplantation is curative but is best used for younger patients who have histocompatible sibling donors. Antithymocyte globulin and cyclosporine restore hematopoiesis in approximately two thirds of patients. However, recovery of blood cell count is often incomplete, recurrent pancytopenia requires retreatment, and some patients develop late complications (especially myelodysplasia). PMID- 11926791 TI - Avoiding nocturnal hypoglycemia: consideration of an extra injection at bedtime. PMID- 11926792 TI - C-reactive protein risk prediction: low specificity, high sensitivity. PMID- 11926793 TI - Welcome, PIER, a new Physicians' Information and Education Resource. PMID- 11926794 TI - Helicobacter pylori eradication therapy for nonulcer dyspepsia. PMID- 11926795 TI - Helicobacter pylori eradication therapy for nonulcer dyspepsia. PMID- 11926796 TI - Helicobacter pylori eradication therapy for nonulcer dyspepsia. PMID- 11926798 TI - Acute exacerbations of chronic obstructive pulmonary disease. PMID- 11926800 TI - Acute exacerbations of chronic obstructive pulmonary disease. PMID- 11926801 TI - Laser removal of radiation tattoos. PMID- 11926802 TI - Putting ethics into the medical record. PMID- 11926806 TI - Summary for patients. Exercise lowers blood pressure. PMID- 11926807 TI - Summary for patients. Splitting the evening insulin dose to avoid low blood sugar levels and to improve sugar control in patients with type 1 diabetes. PMID- 11926808 TI - Summary for patients. Sildenafil (Viagra) may help improve control of pulmonary hypertension. PMID- 11926809 TI - Summary for patients. How a high-fat meal affects blood flow to the heart. PMID- 11926810 TI - Summary for patients. Is C-reactive protein specific for vascular disease in women? PMID- 11926811 TI - Novel antisense and peptide nucleic acid strategies for controlling gene expression. AB - Antisense oligonucleotides have the potential to make revolutionary contributions to basic science and medicine. Oligonucleotides can bind mRNA and inhibit translation. Because they can be rapidly synthesized to be complementary to any sequence, they offer ideal tools for exploiting the massive amount of genome information now available. However, until recently, this potential was largely theoretical, and antisense experiments often produced inconclusive or misleading outcomes. This review will discuss the chemical and biological properties of some of the different types of oligomers now available and describe the challenges confronting in vitro and in vivo use of oligonucleotides. Oligomers with improved chemical properties, combined with advances in cell biology and success in clinical trials, are affording powerful new options for basic research, biotechnology, and medicine. PMID- 11926812 TI - Identification of binding sites for both dsRBMs of PKR on kinase-activating and kinase-inhibiting RNA ligands. AB - The RNA-dependent protein kinase (PKR) is an interferon-induced, RNA-activated enzyme that phosphorylates and inhibits the function of the translation initiation factor eIF-2. PKR has a double-stranded RNA-binding domain (dsRBD) composed of two copies of the dsRNA binding motif (dsRBM). PKR's dsRBD is involved in the regulation of the enzyme as dsRNAs of cellular and viral origins bind to the dsRBD, leading to either activation or inhibition of PKR's kinase activity. In this study, we site-specifically modified each of the dsRBMs of PKR's dsRBD with the hydroxyl radical generator EDTA small middle dotFe and performed cleavage studies on kinase-activating and kinase-inhibiting RNAs. These experiments led to the identification of binding sites for the individual dsRBMs on various RNA ligands including a viral activating RNA (TAR from HIV-1), a viral inhibiting RNA (VA(I) RNA from adenovirus), an aptamer RNA that activates PKR, and a small synthetic inhibiting RNA. These results indicate that some RNAs interact only with one dsRBM, while others can bind both dsRBMs of PKR. In addition, EDTA small middle dotFe modification coupled with site-directed mutagenesis was used to assess the extent of cooperativity in the binding of the two dsRBMs. These experiments support the hypothesis that simultaneous binding of both dsRBMs of PKR occurs on kinase activating RNA ligands. PMID- 11926814 TI - NAIM and site-specific functional group modification analysis of RNase P RNA: magnesium dependent structure within the conserved P1-P4 multihelix junction contributes to catalysis. AB - The tRNA processing endonuclease ribonuclease P contains an essential and highly conserved RNA molecule (RNase P RNA) that is the catalytic subunit of the enzyme. To identify and characterize functional groups involved in RNase P RNA catalysis, we applied self-cleaving ribozyme-substrate conjugates, on the basis of the RNase P RNA from Escherichia coli, in nucleotide analogue interference mapping (NAIM) and site-specific modification experiments. At high monovalent ion concentrations (3 M) that facilitate protein-independent substrate binding, we find that the ribozyme is largely insensitive to analogue substitution and that concentrations of Mg2+ (1.25 mM) well below that necessary for optimal catalytic rate (>100 mM) are required to produce interference effects because of modification of nucleotide bases. An examination of the pH dependence of the reaction rate at 1.25 mM Mg2+ indicates that the increased sensitivity to analogue interference is not due to a change in the rate-limiting step. The nucleotide positions detected by NAIM under these conditions are located exclusively in the catalytic domain, consistent with the proposed global structure of the ribozyme, and predominantly occur within the highly conserved P1-P4 multihelix junction. Several sensitive positions in J3/4 and J2/4 are proximal to a previously identified site of divalent metal ion binding in the P1-P4 element. Kinetic analysis of ribozymes with site-specific N7-deazaadenosine and deazaguanosine modifications in J3/4 was, in general, consistent with the interference results and also permitted the analysis of sites not accessible by NAIM. These results show that, in this region only, modification of the N7 positions of A62, A65, and A66 resulted in measurable effects on reaction rate and modification at each position displayed distinct sensitivities to Mg2+ concentration. These results reveal a restricted subset of individual functional groups within the catalytic domain that are particularly important for substrate cleavage and demonstrate a close association between catalytic function and metal ion-dependent structure in the highly conserved P1-P4 multihelix junction. PMID- 11926813 TI - Poly(C) synthesis by class I and class II CCA-adding enzymes. AB - The CCA-adding enzymes [ATP(CTP):tRNA nucleotidyl transferases], which catalyze synthesis of the conserved CCA sequence to the tRNA 3' end, are divided into two classes. Recent studies show that the class II Escherichia coli CCA-adding enzyme synthesizes poly(C) when incubated with CTP alone, but switches to synthesize CCA when incubated with both CTP and ATP. Because the poly(C) activity can shed important light on the mechanism of the untemplated synthesis of CCA, it is important to determine if this activity is also present in the class I CCA enzymes, which differ from the class II enzymes by significant sequence divergence. We show here that two members of the class I family, the archaeal Sulfolobus shibatae and Methanococcus jannaschii CCA-adding enzymes, are also capable of poly(C) synthesis. These two class I enzymes catalyze poly(C) synthesis and display a response of kinetic parameters to the presence of ATP similar to that of the class II E. coli enzyme. Thus, despite extensive sequence diversification, members of both classes employ common strategies of nucleotide addition, suggesting conservation of a mechanism in the development of specificity for CCA. For the E. coli enzyme, discrimination of poly(C) from CCA synthesis in the intact tRNA and in the acceptor-TPsiC domain is achieved by the same kinetic strategy, and a mutation that preferentially affects addition of A76 but not poly(C) has been identified. Additionally, we show that enzymes of both classes exhibit a processing activity that removes nucleotides in the 3' to 5' direction to as far as position 74. PMID- 11926815 TI - Activation of plant phospholipase Dbeta by phosphatidylinositol 4,5-bisphosphate: characterization of binding site and mode of action. AB - Hydrolysis of phospholipids by plant phospholipase Dbeta (PLDbeta) requires phosphatidylinositol 4,5-bisphosphate [PI(4,5)P2]. Here we show that PLDbeta is stimulated by different polyphosphoinositides, among which PI(4,5)P2 is most effective. On the basis of amino acid sequence analysis, PI(4,5)P2 binding assay, and protein engineering studies, we have identified in the catalytic region of PLDbeta a new PI(4,5)P2 binding region (PBR1), which is conserved in eukaryotic PLDs. PBR1 is a second domain besides the previously characterized N-terminal C2 domain of PLDbeta which also binds PI(4,5)P2. Submillimolar levels of calcium ions, while inhibiting PI(4,5)P2 binding by the C2 domain, enhanced the affinity of PBR1 for that phosphoinositide. Substrate binding by PLDbeta was promoted by PI(4,5)P2-bound PBR1. Isolated, recombinant PBR1 bound PI(4,5)P2 specifically and in a saturable manner. Deletion of PBR1 from PLDbeta or mutation of the conserved basic amino acid residues in PBR1 (K437G/K440G) abolished the enzymatic activity. Circular dichroism spectroscopy revealed a conformational change caused by PI(4,5)P2 binding to the catalytic region of PLD. The conformational change apparently helps in the recruitment of the substrate to the active site of the enzyme. The results taken together allow us to describe an anchorage-scooting model for the synergistic activation of PLDbeta by PI(4,5)P2 and Ca2+. PMID- 11926817 TI - Intermolecular interactions between cholecystokinin-8 and the third extracellular loop of the cholecystokinin-2 receptor. AB - The structure of the third extracellular loop of the human cholecystokinin-2 receptor, CCK2-R(352-379), and its interactions with the C-terminal octapeptide of cholecystokinin (CCK-8) have been determined by high-resolution NMR and computer simulations. In the presence of dodecylphosphocholine micelles, the structure of the receptor fragment consisted of three helices, with the first and third corresponding to residues of the extracellular ends of transmembrane helices (TM) 6 and 7, respectively. The central, extracellular helix, consisting of residues 363-368, was found to be closely associated with the membrane mimetic used during the spectroscopic studies and molecular dynamics (MD) simulations. Upon titration of CCK-8 to the receptor domain, chemical shift perturbation and intermolecular NOEs (Trp30, Met31 of CCK-8 and P371, F374 of CCK2-R) indicated the formation of a stable complex and specific ligand/receptor interactions. Using the NOE-generated intermolecular contact points, extensive MD simulations of CCK-8 bound to the CCK2 receptor were carried out. The results, with CCK-8 in close proximity to TM7, differ from previous structural studies of CCK-8 association with CCK1-R, in which the ligand formed a number of interactions with TM6. These differences may play a role in the ligand specificity displayed by the CCK1 and CCK2 receptor subtypes. PMID- 11926816 TI - Interaction of Asn105 with the retinal chromophore during photoisomerization of pharaonis phoborhodopsin. AB - pharaonis phoborhodopsin (ppR; also called pharaonis sensory rhodopsin II, psR II) is a photoreceptor for negative phototaxis in Natronobacterium pharaonis. ppR has a blue-shifted absorption spectrum with a spectral shoulder, which is highly unique for the archaeal rhodopsin family. The primary reaction of ppR is a cis trans photoisomerization of the retinal chromophore to form the K intermediate, like the well-studied proton pump bacteriorhodopsin (BR). Recent comparative FTIR spectroscopy of the K states in ppR and BR revealed that more extended structural changes take place in ppR than in BR with respect to chromophore distortion and protein structural changes [Kandori, H., Shimono, K., Sudo, Y., Iwamoto, M., Shichida, Y., and Kamo, N. (2001) Biochemistry 40, 9238-9246]. FTIR spectroscopy of the N105D mutant protein reported here assigns the vibrational bands at 1704 and 1700 cm(-1) as C=O stretches of Asn105 in ppR and ppR(K), respectively. A comparative investigation between ppR and BR further reveals that the structure at position 105 in ppR is similar to that of the corresponding position (Asp115) in BR; this observation is supported by the recent X-ray crystallographic structures of ppR [Luecke, H., Schobert, B., Lanyi, J. K., Spudich, E. N., and Spudich, J. L. (2001) Science 293, 1499-1503; Royant, A., Nollert, P., Edman, K., Neutze, R., Landau, E. M., Pebay-Peyroulla, E., and Navarro, J. (2001) Proc. Natl. Acad. Sci. U.S.A. 98, 10131-10136]. Nevertheless, structural changes upon photoisomerization at position 105 in ppR are greater than those at position 115 in BR. As a consequence of a unique chromophore-protein interaction in ppR, extended protein structural changes accompanying retinal photoisomerization occur, and these include Asn105 which is approximately 7 A from the retinal chromophore. PMID- 11926818 TI - Comparison of heat- and pressure-induced unfolding of ribonuclease a: the critical role of Phe46 which appears to belong to a new hydrophobic chain-folding initiation site. AB - To clarify the structural role of Phe46 inside the hydrophobic core of bovine pancreatic ribonuclease A (RNase A), thermal and pressure unfolding of wild-type RNase A and three mutant forms (F46V, F46E, and F46K) were analyzed by fourth derivative UV absorbance spectroscopy. All the mutants, as well as the wild type, exhibited a two-state transition during both thermal and pressure unfolding, and both T(m) and P(m) decreased markedly when Phe46 was replaced with valine, glutamic acid, or lysine. The strongest effect was on the F46K mutant and the weakest on F46V. Both unfolding processes produced identical blue shifts in the fourth-derivative spectra, indicating that the tyrosine residues are similarly exposed in the temperature- and pressure-induced unfolded states. A comparison of Gibbs free energies determined from the pressure and temperature unfoldings, however, gave DeltaG(p)/DeltaG(t) ratios (r) of 1.7 for the wild type and 0.92 +/ 0.03 for the mutants. Furthermore, the DeltaV value for each mutant was larger than that for the wild type. CD spectra and activity measurements showed no obvious major structural differences in the folded state, indicating that the structures of the Phe46 mutants and wild type differ in the unfolded state. We propose a model in which Phe46 stabilizes the hydrophobic core at the boundary between two structural domains. Mutation of Phe46 decreases protein stability by weakening the unfolding cooperativity between these domains. This essential function of Phe46 in RNase A stability indicates that it belongs to a chain folding initiation site. PMID- 11926819 TI - Probing the role of the F-helix in serpin stability through a single tryptophan substitution. AB - Serpins form loop-sheet polymers through the formation of a partially folded intermediate. Through mutagenesis and biophysical analysis, we have probed the conformational stability of the F-helix, demonstrating that it is almost completely unfolded in the intermediate state. The replacement of Tyr160 on the F helix of alpha1-antitrypsin to alanine results in the loss of a conserved hydrogen bond that dramatically reduces the stability of the protein to both heat and solvent denaturation, indicating the importance of Tyr160 in the stability of the molecule. The mutation of Tyr160 to a tryptophan residue, within a fluorescently silent variant of alpha1-antitrypsin, results in a fully active, stable serpin. Fluorescence analysis of the equilibrium unfolding behavior of this variant indicates that the F-helix is highly disrupted in the intermediate conformation. Iodide quenching experiments demonstrate that the tryptophan residue is exposed to a similar extent in both the intermediate and unfolded states. Cumulatively, these data indicate that the F-helix plays an important role in controlling the early conformational changes involved in alpha1 antitrypsin unfolding. The implications of these data on both alpha1-antitrypsin function and misfolding are discussed. PMID- 11926820 TI - Anisotropic dynamics of the JE-2147-HIV protease complex: drug resistance and thermodynamic binding mode examined in a 1.09 A structure. AB - The structure of HIV protease (HIV Pr) bound to JE-2147 (also named AG1776 or KNI 764) is determined here to 1.09 A resolution. This highest-resolution structure for HIV Pr allows refinement of anisotropic displacement parameters (ADPs) for all atoms. Clustering based on the directional information in ADPs defines two sets of subdomains such that within each set, subdomains undergo similar anisotropic motion. These sets are (a) the core of monomer A grouped with both substrate-binding flaps and (b) the core of monomer B coupled to both catalytic aspartates (25A/B). The four-stranded beta-sheet (1-4 A/B and 95-99 A/B) that forms a significant part of the dimer interface exhibits large anisotropic amplitudes that differ from those of the other sets of subdomains. JE-2147 is shown here to be a picomolar inhibitor (K(i) = 41 +/- 18 pM). The structure is used to interpret the mechanism of association of JE-2147, a second-generation inhibitor for which binding is enthalpically driven, with respect to first generation inhibitors for which binding is predominantly entropically driven [Velazquez-Campoy, A., et al. (2001) Arch. Biochem. Biophys. 390, 169-175]. Relative to the entropically driven inhibitor complexes, the JE-2147-HIV Pr complex exhibits an approximately 0.5 A movement of the substrate flaps in toward the substrate, suggesting a more compatible enthalpically driven association. Domains of the protease identified by clustering of ADPs also suggest a model of enthalpy-entropy compensation for all HIV Pr inhibitors in which dynamic coupling of the flaps is offset by an increased level of motion of the beta-sheet domain of the dimer interface (1-4 A/B and 95-99 A/B). PMID- 11926821 TI - Vesicle permeabilization by protofibrillar alpha-synuclein is sensitive to Parkinson's disease-linked mutations and occurs by a pore-like mechanism. AB - Two mutations in the protein alpha-synuclein (A30P and A53T) are linked to an autosomal dominant form of Parkinson's disease. Both mutations accelerate the formation of prefibrillar oligomers (protofibrils) in vitro, but the mechanism by which they promote toxicity is unknown. Protofibrils of wild-type alpha-synuclein bind and permeabilize acidic phospholipid vesicles. This study examines the relative membrane permeabilizing activities of the wild type, mutant, and mouse variants of protofibrillar alpha-synuclein and the mechanism of membrane permeabilization. Protofibrillar A30P, A53T, and mouse variants were each found to have greater permeabilizing activities per mole than the wild-type protein. The leakage of vesicular contents induced by protofibrillar alpha-synuclein exhibits a strong preference for low-molecular mass molecules, suggesting a pore like mechanism for permeabilization. Under conditions in which the vesicular membrane is less stable (lack of calcium as a phospholipid counterion), protofibril permeabilization is less size-selective and monomeric alpha-synuclein can permeabilize via a detergent-like mechanism. We conclude that the pathogenesis of Parkinson's disease may involve membrane permeabilization by protofibrillar alpha-synuclein, the extent of which will be strongly dependent on the in vivo conditions. PMID- 11926822 TI - Spectroscopic characterization of the NO adduct of hydroxylamine oxidoreductase. AB - Hydroxylamine oxidoreductase (HAO) from the autotrophic nitrifying bacterium Nitrosomonas europaea catalyzes the oxidation of NH2OH to NO2-. The enzyme contains eight hemes per subunit which participate in catalysis and electron transport. NO is found to bind to the enzyme and inhibit electron flow to the acceptor protein, cytochrome c554. NO is found to oxidize either partially or fully reduced HAO, but NO will not reduce ferric HAO. Since NO can be reduced but not oxidized to product by HAO, NO is not considered to be a long-lived intermediate in the catalytic mechanism. Substrate oxidation occurs in the presence of bound NO or cyanide, suggesting a second interaction site for substrate with HAO and providing a means for recovery of the NO-inhibited form of the enzyme. Upon addition of NO to oxidized HAO, the integer-spin EPR signal from the active site vanishes, an IR band from NO appears at 1920 cm(-1), and a diamagnetic quadrupole iron doublet appears in Mossbauer spectroscopy with delta = 0.06 mm/s and DeltaEq = 2.1 mm/s. The NO stretching frequency and Mossbauer parameters are characteristic of an [FeNO]6 heme complex. New Mossbauer data on ferric myoglobin-NO are also presented for comparison. The results indicate that NO binds to heme P460 and that the loss of the integer-spin EPR signal is due to the conversion of heme P460 to a diamagnetic S = 0 state and concomitant loss of magnetic interaction with neighboring heme 6. In previous studies where the heme P460-heme 6 interaction was affected by substrate or cyanide binding, a signal attributable to heme 6 was not observable. In contrast, in this work, the NO induced loss of the signal is accompanied by the appearance of a previously unobserved large g(max) (or HALS) low-spin EPR signal from heme 6. PMID- 11926823 TI - FTIR spectroscopic evidence for the involvement of an acidic residue in quinone binding in cytochrome bd from Escherichia coli. AB - In this work, FTIR difference spectroscopy is used to search for possible binding partners and protonable groups involved in the binding of the quinol to cytochrome bd from Escherichia coli. In addition, the electrochemically induced FTIR difference spectra are compared for preparations of the enzyme isolated from cells grown at different oxygen levels in which the quinone content of the membrane is altered. On this basis, difference signals can be tentatively attributed to the vibrational modes of the different quinones types that are associated with the enzyme depending on growth conditions. Furthermore, vibrational modes due to the redox-dependent reorganization of the protein vary depending on the quinone associated with the isolated enzyme. Of particular interest are the observations that a mode at 1738 cm(-1) is decreased and a mode at 1595 cm(-1) is increased as observed in direct comparison to the data obtained from samples grown anaerobically. These signals indicate a change in the protonation state of an aspartic or glutamic acid. Since these changes are observed when the ubiquinone ratio in the preparation increases, the data provide evidence for the modulation of the binding site by the interacting quinone and the involvement of an acidic group in the binding site. The tentative assignments of the vibrational modes are supported by electrochemically induced FTIR difference spectra of cytochrome bd in the presence of the specific quinone binding site inhibitors heptylhydroxyquinoline-N-oxide (HQNO) or 2-methyl-3 undecylquinolone-4. Whereas HQNO leads to strong shifts in the FTIR redox difference spectrum, 2-methyl-3-undecylquinolone-4 induces a specific shift of a mode at 1635 cm(-1), which likely originates from the displacement of the C=O group of the bound quinone. PMID- 11926824 TI - Control of nitric oxide synthase dimer assembly by a heme-NO-dependent mechanism. AB - Homodimer formation is a key step that follows heme incorporation during assembly of an active inducible nitric oxide synthase (iNOS). In cells, heme incorporation into iNOS becomes limited due to interaction between self-generated NO and cellular heme [Albakri, Q., and Stuehr, D. J. (1996) J. Biol. Chem. 271, 5414 5421]. Here we investigated if NO can regulate at points downstream in the process by inhibiting dimerization of heme-containing iNOS monomer. Heme containing monomers were generated by treating iNOS dimer or iNOS oxygenase domain dimer (iNOSoxy) with urea. Both monomers dimerized when incubated with Arg and 6R-tetrahydrobiopterin (H4B), as shown previously [Abu-Soud, H. M., Loftus, M., and Stuehr, D. J. (1995) Biochemistry 34, 11167-11175]. The NO-releasing drug S-nitrosyl-N-acetyl-D,L-penicillamine (SNAP; 0-0.5 mM) inhibited dimerization of iNOS monomer in a dose- and time-dependent manner, without causing heme release. SNAP-pretreated monomer also did not dimerize in response to H4B plus Arg. SNAP converted Arg- and H4B-free iNOS dimer into monomer that could not redimerize, but had no effect on iNOS dimer preincubated with Arg and H4B. Anaerobic spectral analysis showed that NO from SNAP bound to the ferric heme of iNOSoxy monomer or dimer. Adding imidazole as an alternative heme ligand prevented SNAP from inhibiting iNOS monomer dimerization. We conclude that NO and related species can block iNOS dimerization at points downstream from heme incorporation. The damage to heme-containing monomer results from a reaction with the protein and appears irreversible. Although dimeric structure alone does not protect, it does enable Arg and H4B to bind and protect. Inhibition appears mediated by NO coordinating to the ferric heme iron of the monomer. PMID- 11926825 TI - Relaxation kinetics of cytochrome P450 reductase: internal electron transfer is limited by conformational change and regulated by coenzyme binding. AB - The kinetics of internal electron transfer in human cytochrome P450 reductase have been studied using temperature-jump relaxation spectroscopy. Temperature perturbation of CPR reduced at the two-electron level with NADPH yields biphasic absorption transients at 450 and 600 nm. The observed rate, 1/tau, for the fast phase is 2200 +/- 300 s(-1). The absence of this phase in fluorescence transients and in absorption transients collected with dithionite-reduced enzyme indicates this phase does not report on electron/hydride transfer and is consistent with its origin in local conformational change in the vicinity of the FAD isoalloxazine ring. The slow phase (1/tau = 55 +/- 2 s(-1)) observed in the absorption transients obtained with CPR reduced at the two-electron level with NADPH reports on internal electron transfer: FAD(sq)-FMN(sq) --> FAD(ox)-FMN(hq). The observed rate of this transient is slower (1/tau = 11 +/- 0.5 s(-1)) in CPR reduced to the two-electron level by dithionite rather than NADPH, demonstrating that coenzyme binding has an important influence on the observed rate of internal electron transfer. Temperature perturbation experiments with CPR reduced with 10 fold molar excess of NADPH produce monophasic absorption transients (1/tau = 20 +/- 0.2 s(-1)) reporting on internal electron transfer: FAD(sq)-FMN(hq) --> FAD(hq)-FMN(sq). The observed rate constants for electron transfer are substantially less than those expected from analysis of CPR by electron-transfer theory (approximately 10(10) s(-1)). Potential gating mechanisms have been investigated using the temperature-jump method. Observed rates for electron transfer were unaffected in experiments performed in deuterated solvent, indicating that deprotonation does not gate the reaction. Introduction of glycerol into the sample significantly decreased the observed rate for internal electron transfer, suggesting conformational gating of the reaction. Replacement of Trp-676 with His-676 reduces approximately 2-fold the observed rate of internal electron transfer in two-electron-reduced enzyme, whereas the observed rate for FAD(sq)-FMN(hq) --> FAD(hq)-FMN(sq) transfer is increased approximately 13-fold in the W676H mutant reduced with a 10-fold molar excess of NADPH. The studies reveal altered redox properties of the FAD in W676H CPR. The data are discussed in the context of previous stopped-flow studies of human CPR and the X ray crystallographic structure of rat CPR. PMID- 11926826 TI - Beyond the proton abstracting role of Glu-376 in medium-chain acyl-CoA dehydrogenase: influence of Glu-376-->Gln substitution on ligand binding and catalysis. AB - The active site residue, Glu-376, of medium-chain acyl-CoA dehydrogenase (MCAD) has been known to abstract the alpha-proton from acyl-CoA substrates during the course of the reductive half-reaction. The site-specific mutation of Glu-376- >Gln(E376Q) slows down the octanoyl-CoA-dependent reductive half-reaction of the enzyme by about 5 orders of magnitude due to impairment in the proton-transfer step. To test whether the carboxyl group of Glu-376 exclusively serves as the active site base (for abstracting the alpha-proton) during the enzyme catalysis, we undertook a detailed kinetic investigation of the enzyme-ligand interaction and enzyme catalysis, utilizing octanoyl-CoA/octenoyl-CoA as a physiological substrate/product pair and the wild-type and E376Q mutant enzymes as the catalysts. The transient kinetic data revealed that the E376Q mutation not only impaired the rate of octanoyl-CoA-dependent reduction of the enzyme-bound FAD, but also impaired the association and dissociation rates for the binding of the reaction product, octenoyl-CoA. Besides, the E376Q mutation correspondingly impaired the kinetic profiles for the quenching of the intrinsic protein fluorescence during the course of the above diverse (i.e., "chemistry" versus "physical interaction") processes. A cumulative account of the experimental data led to the suggestion that the carboxyl group of Glu-376 of MCAD is intimately involved in modulating the microscopic environment (protein conformation) of the enzyme's active site during the course of ligand binding and catalysis. Arguments are presented that the electrostatic interactions among Glu-376, FAD, and CoA ligands are responsible for structuring the enzyme's active site cavity in the ground and transition states of the enzyme during the above physicochemical processes. PMID- 11926827 TI - Effects of heme ligand mutations including a pathogenic variant, H65R, on the properties of human cystathionine beta-synthase. AB - Human cystathionine beta-synthase is a hemeprotein that catalyzes a pyridoxal phosphate (PLP)-dependent condensation of serine and homocysteine into cystathionine. Biophysical characterization of this enzyme has led to the assignment of the heme ligands as histidine and cysteinate, respectively, which has recently been confirmed by crystal structure determination of the catalytic core of the protein. Using site-directed mutagenesis, we confirm that C52 and H65 represent the thiolate and histidine ligands to the heme. Conversion of C52 to alanine or serine results in spectral properties of the resulting hemeprotein that are consistent with the loss of a thiolate ligand. Thus, the Soret peak blue shifts from 428 to 415 and 417 nm in the ferric forms of the C52S and C52A mutants, respectively, and from 450 to 423 nm in the ferrous states of both mutants. Addition of CO to the dithionite-reduced ferrous C52 mutants results in spectra with Soret peaks at 420 nm. EPR spectroscopy of the ferric C52 variants reveals the predominance of a high-spin species. The H65R mutant, a variant described in a homocystinuric patient, has Soret peaks at 424, 421, and 420 nm in the ferric, ferrous, and ferrous CO states, respectively. EPR spectroscopy reveals predominance of the low-spin species. Both C52A and C52S mutations lead to protein with substoichiometric heme (19% with respect to wild type); however, the PLP content is comparable to that of wild-type enzyme. The heme and PLP contents of the H65R mutant are 40% and 75% that of wild-type enzyme. These results indicate that heme saturation does not dictate PLP saturation in these mutant enzymes. Both H65 and C52 variants display low catalytic activity, revealing that changes in the heme binding domain modulate activity, consistent with a regulatory role for this cofactor. PMID- 11926829 TI - Site-specific cross-linking reveals a differential direct interaction of class 1, 2, and 3 ADP-ribosylation factors with adaptor protein complexes 1 and 3. AB - We have used a site-specific photo-cross-linking approach to identify direct interactions between clathrin adaptor protein (AP)1 complexes and small GTPases of the ADP-ribosylation factor (ARF) family and to explore the specificity of this interaction on immature secretory granule (ISG) membranes. ISG membranes are a well-characterized, highly enriched preparation of membranes that has previously been shown to have the membrane-associated factors for ARF1 recruitment that are not present on artificial liposomes. All three classes of ARF proteins could be recruited to ISG membranes, displaying differential requirements for GTPgammaS. We found that ARF1, ARF5, and ARF6 interacted directly with the beta1-adaptin subunit of AP-1 in the presence of GTPgammaS. Furthermore, we observed a direct interaction between the switch 1 region of ARF1 and the N-terminal trunk domains of gamma- and beta1-adaptin. In addition, both ARF1 and ARF6 but not ARF5 interacted directly with the beta3- and delta-adaptin subunits of AP-3. No interaction was observed between AP-2 and any of the ARF proteins. Our results delineate the specificity and provide evidence of a direct interaction between different ARF proteins and the AP complexes AP-1 and AP-3 on natural ISG membranes and show that residues in the switch 1 region of ARF proteins can selectively bind to the trunk domains of these complexes. PMID- 11926828 TI - Kinetic and magnetic resonance studies of the role of metal ions in the mechanism of Escherichia coli GDP-mannose mannosyl hydrolase, an unusual nudix enzyme. AB - Escherichia coli GDP-mannose mannosyl hydrolase (GDPMH), a homodimer, catalyzes the hydrolysis of GDP-alpha-D-sugars to yield the beta-D-sugar and GDP by nucleophilic substitution with inversion at the C1' carbon of the sugar [Legler, P. M., Massiah, M. A., Bessman, M. J., and Mildvan, A. S. (2000) Biochemistry 39, 8603-8608]. GDPMH requires a divalent cation for activity such as Mn2+ or Mg2+, which yield similar kcat values of 0.15 and 0.13 s(-1), respectively, at 22 degrees C and pH 7.5. Kinetic analysis of the Mn2+-activated enzyme yielded a K(m) of free Mn2+ of 3.9 +/- 1.3 mM when extrapolated to zero substrate concentration (K(a)Mn2+), which tightened to 0.32 +/- 0.18 mM when extrapolated to infinite substrate concentration (K(m)Mn2+). Similarly, the K(m) of the substrate extrapolated to zero Mn2+ concentration (K(S)(GDPmann) = 1.9 +/- 0.5 mM) and to infinite Mn2+ concentration (K(m)(GDPmann) = 0.16 +/- 0.09 mM) showed an order of magnitude decrease at saturating Mn2+. Such mutual tightening of metal and substrate binding suggests the formation of an enzyme-metal-substrate bridge complex. Direct Mn2+ binding studies, monitoring the concentration of free Mn2+ by EPR and of bound Mn2+ by its enhanced paramagnetic effect on the longitudinal relaxation rate of water protons (PRR), detected three Mn2+ binding sites per enzyme monomer with an average dissociation constant (K(D)) of 3.2 +/- 1.0 mM, in agreement with the kinetically determined K(a)Mn2+. The enhancement factor (epsilon(b)) of 11.5 +/- 1.2 indicates solvent access to the enzyme-bound Mn2+ ions. No cross relaxation was detected among the three bound Mn2+ ions, suggesting them to be separated by at least 10 A. Such studies also yielded a weak dissociation constant for the binary Mn2+-GDP-mannose complex (K1 = 6.5 +/- 1.0 mM) which significantly exceeded the kinetically determined K(m) values of Mn2+, indicating the true substrate to be GDP-mannose rather than its Mn2+ complex. Substrate binding monitored by changes in 1H-15N HSQC spectra yielded a dissociation constant for the binary E-GDP-mannose complex (K(S)(GDPmann)) of 4.0 +/- 0.5 mM, comparable to the kinetically determined K(S) value (1.9 +/- 0.5 mM). To clarify the metal stoichiometry at the active site, product inhibition by GDP, a potent competitive inhibitor (K(I) = 46 +/- 27 microM), was studied. Binding studies revealed a weak, binary E-GDP complex (K(D)(GDP) = 9.4 +/- 3.2 mM) which tightened approximately 500-fold in the presence of Mn2+ to yield a ternary E Mn2+-GDP complex with a dissociation constant, K3(GDP) = 18 +/- 9 microM, which overlaps with the K(I)(GDP). The tight binding of Mn2+ to 0.7 +/- 0.2 site per enzyme subunit in the ternary E-Mn2+-GDP complex (K(A)' = 15 microM) and the tight binding of GDP to 0.8 +/- 0.1 site per enzyme subunit in the ternary E-Mg2+ GDP complex (K3 < 0.5 mM) indicate a stoichiometry close to 1:1:1 at the active site. The decrease in the enhancement factor of the ternary E-Mn2+-GDP complex (epsilon(T) = 4.9 +/- 0.4) indicates decreased solvent access to the active site Mn2+, consistent with an E-Mn2+-GDP bridge complex. Fermi contact splitting (4.3 +/- 0.2 MHz) of the phosphorus signal in the ESEEM spectrum established the formation of an inner sphere E-Mn2+-GDP complex. The number of water molecules coordinated to Mn2+ in this ternary complex was determined by ESEEM studies in D2O to be two fewer than on the average Mn2+ in the binary E-Mn2+ complexes, consistent with bidentate coordination of enzyme-bound Mn2+ by GDP. Kinetic, metal binding, and GDP binding studies with Mg2+ yielded dissociation constants similar to those found with Mn2+. Hence, GDPMH requires one divalent cation per active site to promote catalysis by facilitating the departure of the GDP leaving group, unlike its homologues the MutT pyrophosphohydrolase, which requires two, or Ap4A pyrophosphatase, which requires three. PMID- 11926830 TI - The HIV-1 gp41 N-terminal heptad repeat plays an essential role in membrane fusion. AB - For many different enveloped viruses the crystal structure of the fusion protein core has been established. A striking conservation in the tertiary and quaternary arrangement of these core structures is repeatedly revealed among members of diverse families. It has been proposed that the primary role of the core involves structural rearrangements which facilitate apposition between viral and target cell membranes. Forming the internal trimeric coiled coil of the core, the N terminal heptad repeat (NHR) of HIV-1 gp41 was suggested to have additional roles, due to its ability to bind biological membranes. The NHR is adjacent to the N-terminal hydrophobic fusion peptide (FP), which alone can fuse biological membranes. To investigate the role of the NHR in membrane fusion, we synthesized and functionally characterized HIV-1 gp41 peptides corresponding to the FP and NHR alone, as well as continuous peptides made of both FP and NHR (wild type and mutant). We show here that a consecutive, 70-residue peptide consisting of both the FP and NHR (gp41/1-70) has dramatic fusogenic properties. The effect of including the complete NHR, as compared to shorter 23-, 33-, or 52-residue N terminal peptides, is illustrated by a leap in lipid mixing of phosphatidylcholine (PC) large unilamellar vesicles (LUV) and clearly delineates the synergistic role of the NHR in the fusion event. Furthermore, a mutation in the NHR that renders the virus noninfectious is reflected by a significant reduction in in vitro lipid mixing induced by the mutant, gp41/1-70 (I62D). Additional spectroscopic studies, characterizing membrane binding and apposition induced by the peptides, help to clarify the role of the NHR in membrane fusion. PMID- 11926831 TI - Properties and utility of the peculiar mixed disulfide in the bacterial glutathione transferase B1-1. AB - Bacterial glutathione transferases appear to represent an evolutionary link between the thiol:disulfide oxidoreductase and glutathione transferase superfamilies. In particular, the observation of a mixed disulfide in the active site of Proteus mirabilis glutathione transferase B1-1 is a feature that links the two families. This peculiar mixed disulfide between Cys10 and one GSH molecule has been studied by means of ESR spectroscopy, stopped-flow kinetic analysis, radiochemistry, and site-directed mutagenesis. This disulfide can be reduced by dithiothreitol but even a thousand molar excess of GSH is poorly effective due to an unfavorable equilibrium constant of the redox reaction (K(eq) = 2 x 10(-4)). Although Cys10 is partially buried in the crystal structure, in solution it reacts with several thiol reagents at a higher or comparable rate than that shown by the free cysteine. Kinetics of the reaction of Cys10 with 4,4' dithiodipyridine at variable pH values is consistent with a pK(a) of 8.0 +/- 0.1 for this residue, a value about 1 unit lower than that of the free cysteine. The 4,4'-dithiodipyridine-modified enzyme reacts with GSH in a two-step mechanism involving a fast precomplex formation, followed by a slower chemical step. The natural Cys10-GSH mixed disulfide exchanges rapidly with free [3H]GSH in a futile redox cycle in which the bound GSH is continuously replaced by the external GSH. Our data suggest that the active site of the bacterial enzyme has intermediate properties between those of the recently evolved glutathione transferases and those of the thiol:disulfide oxidoreductase superfamily. PMID- 11926832 TI - Islet amyloid: phase partitioning and secondary nucleation are central to the mechanism of fibrillogenesis. AB - Islet amyloid polypeptide (IAPP) contributes to the pathogenesis of type II diabetes by depositing as cytotoxic amyloid fibers in the endocrine pancreas. Fiber formation occurs with a marked conformational change from an unstructured precursor. Using real-time quantitative kinetic methods, fibrillogenesis was characterized as a function of protein, denaturant, and seed concentration. Several observations are in sharp contrast to the expectations for nucleation dependent polymerization. First, the half-time of conversion for both de novo and seeded kinetics were found to be independent of protein concentration. Second, while elongation kinetics scale linearly with protein concentration, they are relatively insensitive to changes in the total seed concentration. Third, seeded bypass of de novo fiber formation kinetics shows a lag phase. The seeded lag phase is eliminated by a time delay before the introduction of seed to a de novo reaction. Last, conversion is highly cooperative, with the time required for 10 90% conversion occurring much faster than the lag time. At a minimum, four kinetic steps are required to describe these observations: activation, fiber independent nucleation, fiber-dependent nucleation, and elongation. Furthermore, we invoke a phase transition in which protein initially forms an off-pathway dispersion. This single construct allows us to model both the concentration independence of the de novo reaction time and the first-order concentration dependence of the elongation kinetics. Marked acceleration of this reaction by hexafluoro-2-propanol reinforces this view by altering the relative solubility of the two phases and/or by stabilizing hydrogen-bonded structures in the transition states of the reaction pathway. PMID- 11926833 TI - Cooperation between lateral ligand mobility and accessibility for receptor recognition in selectin-induced cell rolling. AB - Selectin-induced leukocyte rolling along the endothelial surface is an essential step in the immune response. Several in vitro studies showed that this cell rolling is a highly regulated adhesion phenomenon, controlled by the kinetics and forces of selectin-ligand interactions. In the flow chamber study presented here, we focused on the requirements on the ligand structure in this context. A series of neoglycolipids bearing the binding epitope Sialyl Lewis X was synthesized and used as artificial ligands. These lipids differed in their spacer structures between headgroup and membrane anchor, resulting in a gradual variation in accessibility and mobility of the binding epitope when immobilized in model membranes. Consequently, analysis of cell rolling along such membranes allowed correlation of ligand structures and functionality. All model membranes containing such ligands were further characterized by film balance measurements, epifluorescence, and atomic force microscopy. Generally, the glycolipids exhibited a high tendency for lateral aggregation, but the resulting clusters were of different morphology. This was also reflected by strong differences in the rolling experiments. Our results confirm that, in addition to a sufficient headgroup accessibility, the cell rolling process is governed by two further interdependent factors: (i) the headgroup flexibility caused by the intramolecular uncoupling between the headgroup and the hydrophobic moiety due to introduction of a spacer, and (ii) the stiffness of the molecules resulting from their supramolecular arrangement in clustered assemblies. Since both factors are influenced simultaneously by the spacer modification, we present for the first time a clear correlation between structural aspects of selectin ligands and their ability to mediate cell rolling. This might help to develop a better understanding for the function of the natural selectin ligands. PMID- 11926834 TI - Folding kinetics of the protein pectate lyase C reveal fast-forming intermediates and slow proline isomerization. AB - Pectate lyase C (pelC) is a member of the class of proteins that possess a parallel beta-helix folding motif. A study of the kinetic folding mechanism is presented in this report. Kinetic circular dichroism (CD) and fluorescence have been used to observe changes in the structure of pelC as a function of time upon folding and unfolding. Three folding phases are observed with far-UV CD and four phases are observed with near-UV CD. The two slowest phases have relaxation times on the order of 21 and 46 s in aqueous buffer. Double-jump refolding assays and the measured activation enthalpies (16.0 and 21.2 kcal/mol for the respective slow phases) suggest that these two phases are the result of the slow cis-trans isomerization of prolyl-peptide bonds. We have determined that the earliest observed folding phase involves the formation of most, if not all, of the secondary structure with a relaxation time of 0.25 s. We also observed a phase by near-UV CD on the order of 0.25 s. This suggests that along with the appearance of secondary structure, some tertiary contacts are made. There is one kinetic phase observed in the near-UV CD and fluorescence that has no corresponding far UV CD phase. This occurs with a relaxation time of 1.1 s. The temperature dependence of the natural log of the folding rate constant suggests that folding occurs via a sequential mechanism in which an on-pathway intermediate in rapid equilibrium with the unfolded protein is present. Semiempirical CD calculations support the idea that the beta-helix region of pelC forms in the fast kinetic phase, yielding near-native secondary and tertiary structures in that region. This is followed by the slower formation of the loop regions connecting individual strands of the beta-helix. PMID- 11926835 TI - Identification of proline residues responsible for the slow folding kinetics in pectate lyase C by mutagenesis. AB - The folding mechanism of pectate lyase C (pelC) involves two slow phases that have been attributed to proline isomerization. To have a more detailed and complete understanding of the folding mechanism, experiments have been carried out to identify the prolyl-peptide bonds responsible for the slow kinetics. Site directed mutagenesis has been used to mutate each of the prolines in pelC to alanine or valine. It has been determined that isomerization of the Leu219-Pro220 peptide bond is responsible for the slowest folding phase observed. The mutant P220A shows kinetic behavior that is identical to the wild-type protein except that the 46-s phase is eliminated. The Leu219-Pro220 peptide bond is cis in the native enzyme. An analysis of the free energy of unfolding of this mutant indicates that the mutation destabilizes the protein by about 4 kcal/mol. However, it appears that the major refolding pathways are unaltered. Further mutations were carried out in order to assign the peptide bond responsible for the 21-s folding phase in pelC. Mutation of the remaining 11 prolines, which are trans in the native enzyme, resulted in no significant changes in the kinetic folding behavior. The conclusion from these experiments is that the 21-s phase involves isomerization of more than one prolyl-peptide bond with similar activation energies. PMID- 11926836 TI - Mechanistic aspects of the covalent flavoprotein dimethylglycine oxidase of Arthrobacter globiformis studied by stopped-flow spectrophotometry. AB - Dimethylglycine oxidase (DMGO) is a covalent flavoenzyme from Arthrobacter globiformis that catalyzes the oxidative demethylation of dimethylglycine to yield sarcosine, formaldehyde, and hydrogen peroxide. Stopped-flow and steady state kinetic studies have been used to study the reductive and oxidative half reactions using dimethylglycine and O2 as substrates. The reductive half-reaction is triphasic. The rate of the fast phase is dependent on substrate concentration, involves flavin reduction, and has a limiting rate constant of 244 s(-1). This phase also displays a kinetic isotope effect of 2.9. Completion of the first kinetic phase generates an intermediate with broad spectral signature between 350 and 500 nm, which is attributed to a reduced enzyme-iminium charge-transfer species, similar to the purple intermediate that accumulates in reactions of D amino acid oxidase (DAAO) with alanine. The second phase (16 s(-1)) is independent of substrate concentration and is attributed to iminium hydrolysis/deprotonation. The third phase (2 s(-1)) is attributed to product release, the rate of which is less than the steady-state turnover rate (10.6 s( 1)). Flavin oxidation of dithionite- and dimethylglycine-reduced enzyme by O2 occurs in a single phase, and the rate shows a linear dependence on oxygen concentration, giving bimolecular rate constants of 342 and 201 mM(-1) x s(-1), respectively. Enzyme-monitored turnover experiments indicate that decay of the reduced enzyme-iminium intermediate is rate-limiting, consistent with rate constants determined from single turnover studies. A minimal kinetic mechanism is presented, which establishes a close relationship to the mechanism of action of DAAO. The covalent flavin in dimethylglycine oxidase is identified as an alphaN1 histidyl48-FAD, and equilibrium titration studies establish a single redox center that displays typical flavoprotein 'oxidase' characteristics. PMID- 11926837 TI - Proximity of bound Hoechst 33342 to the ATPase catalytic sites places the drug binding site of P-glycoprotein within the cytoplasmic membrane leaflet. AB - The P-glycoprotein multidrug transporter carries out ATP-driven cellular efflux of a wide variety of hydrophobic drugs, natural products, and peptides. Multiple binding sites for substrates appear to exist, most likely within the hydrophobic membrane spanning regions of the protein. Since ATP hydrolysis is coupled to drug transport, the spatial relationship of the drug binding sites relative to the ATPase catalytic sites is of considerable interest. We have used a fluorescence resonance energy transfer (FRET) approach to estimate the distance between a bound substrate and the catalytic sites in purified P-glycoprotein. The fluorescent dye Hoechst 33342 (H33342), a high-affinity P-glycoprotein substrate, bound to the transporter and acted as a FRET donor. H33342 showed greatly enhanced fluorescence emission when bound to P-glycoprotein, together with a substantial blue shift, indicating that the drug binding site is located in a nonpolar environment. Cys428 and Cys1071 within the catalytic sites of P glycoprotein were covalently labeled with the acceptor fluorophore NBD-Cl (7 chloro-4-nitrobenz-2-oxa-1,3-diazole). H33342 fluorescence was highly quenched when bound to NBD-labeled P-glycoprotein relative to unlabeled protein, indicating that FRET takes place from the bound dye to NBD. The distance separating the bound dye from the NBD acceptor was estimated to be approximately 38 A. Transition-state P-glycoprotein with the complex ADP*orthovanadate*Co2+ stably trapped at one catalytic site bound H33342 with similar affinity, and FRET measurements led to a similar separation distance estimate of 34 A. Since previous FRET studies indicated that a fluorophore bound within the catalytic site was positioned 31-35 A from the interfacial region of the bilayer, the H33342 binding site is likely located 10-14 A below the membrane surface, within the cytoplasmic leaflet of the membrane, in both resting-state and transition state P-glycoprotein. PMID- 11926838 TI - Associations between maternal and child health status and patterns of medical care use. AB - OBJECTIVE: To determine associations in health status and health care utilization between mothers and their children. METHODS: Cross-sectional analysis of the 1996 97 Community Tracking Study Household Survey. Separate logistic regression models describing each type of service use were conducted and adjusted for child age, health status, mother age, race, education, and respondent type. Models incorporated maternal health care use, employment, poverty, child's insurance status, and family type as independent variables. RESULTS: Sample included 9803 mother-child pairs, for a weighted sample of 35 651 048 pairs. Compared to mothers reporting excellent health, mothers reporting very good to good health status and fair or poor health were more likely to have children in good, fair, or poor health (odds ratio 2.00, 95% confidence interval 1.63-2.45; and odds ratio 4.16, 3.36-5.15, respectively). For each of 5 types of service use, mothers' and children's use were strongly associated: 1+ physician visits (2.42, 2.07-2.84); 6+ physician visits (2.07, 1.80-2.37); emergency department use (2.01, 1.75-2.31); hospitalizations (1.56, 1.10-2.22); and mental health visits (7.07, 5.67-8.82). CONCLUSIONS: Associations in health service use were noted across a broad array of services for women and their children. These associations may reflect similar tendencies to seek care and suggest the need to consider patterns of maternal use in trying to understand and improve patterns of health care utilization for children. PMID- 11926839 TI - Specialty referrals made during telephone conversations with parents: a study from the pediatric research in office settings network. AB - OBJECTIVES: To characterize variation in pediatricians' telephone referral practices, to identify differences in the types of referrals made during telephone versus office visit encounters, and to examine the impact of referring by telephone on coordination and outcomes of the referral as assessed by physicians. METHODS: We conducted a prospective study of a consecutive sample of referrals (N = 1856) made from the offices of 142 pediatricians in a national practice-based research network. During 20 consecutive practice-days, physicians completed questionnaires about patients referred during regular business hours. They used office records 3 months later to complete questionnaires about referral outcomes. RESULTS: Pediatricians made 1 telephone referral every 5 practice-days, which constituted 27.5% of all referrals they made during office hours. Pediatricians who saw more patients per day, saw more patients in gatekeeping health plans, and referred more during office visits made more telephone referrals than their counterparts. Compared with specialty referrals made during office visits, those occurring during telephone encounters were more frequently at the request of parents or because of insurance administrative guidelines. Office visit referrals were more often made for diagnostic evaluation or a surgical procedure. Referrals made during telephone conversations were less well coordinated: office staff or referring physicians scheduled fewer specialty appointments and were less likely to send information to specialists. Three months after referrals were made, specialist feedback and referring physician satisfaction with specialty care were comparable between the two groups. CONCLUSIONS: Specialty referrals made during telephone conversations with patients are a regular occurrence in pediatric practice. Changes in the health system that lead to greater demands on primary care physician productivity or more patients in gatekeeping health plans will likely increase the number of referrals made during telephone conversations with parents. Pediatricians are less likely to coordinate telephone referrals than office visit referrals. Pediatricians are frequently unaware whether or not referrals are completed. PMID- 11926840 TI - Use of complementary/alternative therapies among children in primary care pediatrics. AB - OBJECTIVES: To determine the prevalence of and factors associated with use of complementary/alternative therapies (CAM) by pediatric patients seeking primary care. DESIGN AND SETTING: A self-report questionnaire was administered to parents/caregivers in 6 general pediatric practices in urban and suburban Detroit from August 1999 to December 1999. RESULTS: A total of 1013 questionnaires were completed; 67.5% of the patients were 5 years of age or younger. The overall use of CAM was 12%. Factors in families associated with use of CAM were maternal age greater than 31 years (P =.001), religious affiliation (P =.001), parent/caretaker born outside of the United States (P =.04), and use of CAM by the parent/caretaker or his/her spouse (P =.001). Significant factors associated with the children who used CAM were age greater than 5 years (P =.001), pediatric visit for an illness (P =.05), regular medication use (P =.001), and having an ongoing medical problem (P =.001). The most common types of CAM used were herbs (41%), prayer healing (37%), high-dose vitamin therapy and other nutritional supplements (34.5%), folk/home remedies (28%), massage therapy (19%), and chiropractic (18%). The majority of CAM users (66%) did not report the use of CAM to their primary care physician. A logistic regression analysis revealed that use of CAM by parents/caretakers was the single best predictor of CAM use in a child. CONCLUSION: CAM use is significant among children who visit pediatric practices. Pediatricians should inquire about CAM use among patients, particularly those with ongoing medical problems and those with parents/caretakers who use CAM for themselves. PMID- 11926841 TI - Adolescents' use of complementary and alternative medicine. AB - OBJECTIVE: To examine the prevalence of complementary and alternative medicine (CAM) use in a random sample of adolescents living in Monroe County, New York. METHODS: Questions about use of a variety of CAM therapies in the previous 6 months were asked of adolescents during a random-digit-dial telephone survey. RESULTS: A total of 54% of the 361 adolescents surveyed had used at least 1 CAM remedy. The most frequently used therapies were massage (13.2%), prayer or faith healing (13.1%), herbs (11.5%), megadose vitamins (10.6%), and special exercises (10.1%). Natural performance enhancers were used by 14.7% of the boys, but less than 1% of the girls (P <.001). Use of health care without parental knowledge, time spent in school clubs, and perceived parental use and friend use of CAM were all associated with CAM use in a logistic regression model. CONCLUSION: More than half of the adolescents in this county use CAM therapies, and a significant number use pharmacologically active substances. Physicians treating adolescents should ask adolescents about CAM use. PMID- 11926842 TI - Ambulatory use of psychotropics by employer-insured children and adolescents in a national managed care organization. AB - OBJECTIVE: To provide a description of the ambulatory use of psychotropic medications by children and adolescents in a large, geographically diverse employer-insured population. DESIGN: This retrospective observational study used administrative claims data for 1995-1999 for members under age 20 in 6 Independent Practice Association health plans affiliated with UnitedHealth Group. We calculated the prevalences of use for 4 major therapeutic drug classes: central nervous system stimulants (CNSSs), selective serotonin reuptake inhibitors (SSRIs), tricyclic antidepressants (TCAs), and other antidepressants (OADs). Changes over time by age, gender, geographic region, and prescriber specialty were analyzed across drug classes. RESULTS: The prevalence of CNSS, SSRI, and OAD use steadily increased over the 5-year period, whereas TCA use decreased. The prevalence of use of the most commonly used classes, the CNSS and SSRI classes, increased from 23.8 to 30.0 per 1000 and 7.9 to 12.8, respectively. There was variability across and within geographic regions. Pediatricians were the most frequent first prescribers of CNSS, and psychiatrists were most likely to prescribe SSRIs. CONCLUSION: Acceleration of use of psychotropic medications is slower in an employer-insured national population. Since primary care physicians are frequent prescribers of psychotropics, their training and expertise are crucial. PMID- 11926843 TI - Assessment of suspicion of abuse in the primary care setting. AB - OBJECTIVES: To describe the primary care practitioner's assessment of the likelihood that an injury was caused by physical abuse. The hypotheses were 1) practitioners face great uncertainty as to the possibility that an injury may have been caused by abuse; a measure that assigns variable degrees of suspicion to childhood injuries can be developed that will reveal this uncertainty; and 2) practitioner factors and patient factors influence this suspicion. METHODS: Primary care practitioners in a regional practice-based research network prospectively collected information about each consecutive office encounter during a 4-week study period. For injury-related visits, the practitioner described injury type, reported cause and severity, and the practitioner's assessment of the cause of injury. Practitioners also used a 5-point Suspicion Scale to identify their level of suspicion that the injury was caused by abuse, with 1 equating to impossible and 5 equating to virtually certain. A subset of practitioners gave information about child and family risk factors. The practitioner's reporting activity was not studied. RESULTS: Participating practitioners (n = 85) in 17 practices collected information about 12 510 office encounters, including 659 injuries. Although the practitioners assessed no injuries as "caused by abuse," they rated 21% of the injuries as having "some suspicion" of abuse. Practitioners were more likely to have "some suspicion" of abuse for those children who were Hispanic or African-American (vs. White) (P =.001, chi(2)) and for those children whose mothers had no college education (P =.018, chi(2)). In multivariate logistic regression modeling, "some suspicion" of abuse was associated with higher injury severity (odds ratio [OR] 3.4, 95% confidence interval [CI] 1.7, 7.0), age <6 years (OR 2.9, 95% CI 1.5, 5.6), Medicaid or self-pay health care (OR 1.4, 95% CI 1.4, 5.3), practitioner identification of family risk factors (OR 4.8, 95% CI 1.6, 14.6), and more recent practitioner education about child abuse (OR 2.9, 95% CI 1.4, 5.8). CONCLUSION: Primary care practitioners reported some degree of suspicion that 21% of injuries they evaluated were caused by abuse. Patient factors and practitioner factors influenced their suspicion. PMID- 11926844 TI - Urban elementary school personnel's perceptions of student health and student health needs. AB - OBJECTIVE: To determine the perceptions of school personnel with regard to urban students' health and health needs as part of a planning process for development and implementation of school-linked health services. METHODS: A self administered, close-ended questionnaire was given to 294 schoolteachers and school staff to assess school personnel perceptions of 1) health needs of students, 2) priority of need for health services or programs delivered on-site or off-site, and 3) selected factors suspected to affect implementation of new school health services. RESULTS: A total of 72% of teachers and 77% of staff completed the survey. The most highly ranked need by school personnel was mental health, followed by violence and anger management and physical abuse and domestic violence. The most commonly requested services on-site and off-site were for physical abuse or domestic violence, mental health, and drug or other substance abuse. Only 40% of teachers and staff rated current health services as excellent or good. A total of 72% of teachers and staff had some level of discomfort with their current responsibilities regarding the health of students. Parental involvement was reported to be quite low, with only 9% of teachers and staff rating it as excellent or good. CONCLUSIONS: he greatest health care need identified by these elementary school personnel was not for management of chronic or acute physical health conditions, but for mental health services and programs to meet the emotional and psychosocial needs of students. When implementing school-linked or school-based mental health services, focus needs to be placed on 3 areas: 1) education of school health and educational personnel regarding common mental health issues, 2) development of systems that improve knowledge of and connection with mental health resources inside and outside the school, and 3) development of systems that improve communication within the school and between the school and parents regarding health services. PMID- 11926845 TI - Resident preparedness for practice: a longitudinal cohort study. AB - OBJECTIVE: To determine whether the perception of preparedness for practice changes over time. DESIGN: Questionnaire survey of University of Massachusetts residents 5 years after an initial survey. Responses to individual questions in the 2 surveys were compared for each graduate and the Wilcoxon rank sum test applied. A supplementary questionnaire addressed current confidence in areas with relatively low scores in both surveys. RESULTS: All 24 eligible graduates responded. The high rating of overall sense of preparedness was identical in the 2 surveys. Differences were statistically significant in only 3 categories: common illnesses, office gastroenterology, and office gynecology-all from a lower estimate of preparedness initially to a higher estimate in retrospect. Six areas continued to receive relatively low scores: nutrition, patient scheduling, cost effectiveness, telephone management, office gynecology, and office orthopedics. Respondents feel more confident currently with nutrition, patient scheduling, and telephone management but not with cost-effectiveness, gynecology, or orthopedics. CONCLUSIONS: Residents paired with office-based practitioners for their continuity experience report feeling well prepared for practice both on practice entry and 5 to 9 years later. In the 6 areas of relatively low preparedness, experience improved confidence with nutrition, patient scheduling, and telephone management, but not cost-effectiveness, gynecology, or orthopedics. The hypothesis that clinical areas of relative weakness at the end of residency may remain so years later deserves to be tested. PMID- 11926847 TI - Plasma D-dimers in the diagnosis of venous thromboembolism. AB - Clinical suspicion for venous thromboembolism (VTE) mandates objective testing to confirm or exclude the diagnosis. However, current imaging modalities are imperfect because of a small but important risk of complications with invasive techniques or limited sensitivity with noninvasive ones. A diagnostic tool for VTE is needed that is noninvasive and highly accurate, allowing immediate treatment decisions to be made in most cases. Plasma D-dimers (D-ds), specific cross-linked fibrin derivatives, partially fulfill these criteria in that they are sensitive markers for thrombosis but lack specificity. They therefore cannot be used to make a positive diagnosis of VTE; however, they generally have high negative predictive value and are useful as an exclusionary test, a potentially important role given that VTE is eventually ruled out in most patients investigated. Clinical management studies are clarifying the role of D-ds in the diagnostic paradigm of VTE: negative ultrasound and D-d findings obviate the need for serial imaging in suspected deep vein thrombosis, and anticoagulant therapy can be safely withheld in patients with non-high clinical suspicion for pulmonary embolism and non-high probability ventilation perfusion scan if D-d test results are negative. More recently, the combination of a negative SimpliRED (AGEN Biomedical Ltd, Brisbane, Australia) D-d result and low clinical suspicion derived using a formal scoring system has been shown to exclude deep vein thrombosis and pulmonary embolism and to obviate the need for imaging. Several different D-d assays are now available, and clinicians should be aware of the performance characteristics of the test used before incorporation into diagnostic algorithms as these will differ between assays, and the results of clinical management studies cannot necessarily be safely extrapolated to assays other than those specifically evaluated. If alternative assays are to be substituted, these should consistently have been shown to possess equivalent or greater sensitivity. PMID- 11926848 TI - Primary care quality in the Medicare Program: comparing the performance of Medicare health maintenance organizations and traditional fee-for-service medicare. AB - BACKGROUND: Since 1972, Medicare beneficiaries have had the option of enrolling in a Medicare-qualified health maintenance organization (HMO). Little information exists to inform beneficiaries' choices between the traditional fee-for-service (FFS) Medicare program and an HMO. OBJECTIVES: To compare the primary care received by seniors in Medicare HMOs with that of seniors in the traditional FFS Medicare program, and among HMOs, and to examine performance differences associated with HMO model-type and profit status. METHODS: Data were derived from a cross-sectional observational survey of Medicare beneficiaries 65 years or older in the 13 states with mature, substantial Medicare HMO markets. Only beneficiaries continuously enrolled for 12 months or more in traditional FFS Medicare or a qualified Medicare HMO were eligible. Data were obtained using a 5 stage protocol involving mail and telephone (64% response rate). Analyses included respondents who identified a primary physician and had all required data elements (N = 8828). We compared FFS and HMO performance on 11 summary scales measuring 7 defining characteristics of primary care: (1) access, (2) continuity, (3) integration, (4) comprehensiveness, (5) "whole-person" orientation, (6) clinical interaction, and (7) sustained clinician-patient partnership. RESULTS: For 9 of 11 indicators, performance favored traditional FFS Medicare over HMOs (P<.001). Financial access favored HMOs (P<.001). Preventive counseling did not differ by system. Network-model HMOs performed more favorably than staff/group model HMOs on 9 of 11 indicators (P<.001). Few differences were associated with HMO profit status. CONCLUSIONS: The findings are consistent with previous comparisons of indemnity insurance and network-model and staff/group-model HMOs in elderly and nonelderly populations. The stability of results across time, geography, and populations suggests that the relative strengths and weaknesses of each system are enduring attributes of their care. Medicare enrollees seem to face the perennial cost-quality trade-off: that is, deciding whether the advantages of primary care under traditional FFS Medicare are worth the higher out-of-pocket costs. PMID- 11926849 TI - Use and referral patterns for 22 complementary and alternative medical therapies by members of the American College of Rheumatology: results of a national survey. AB - BACKGROUND: This study was designed to determine rheumatologists' self-reported knowledge, perceptions of legitimacy, referral patterns, and use in practice of 22 complementary and alternative medicine (CAM) therapies. METHODS: A survey was mailed to a random sample of 2000 physician members of the American College of Rheumatology asking respondents which (if any) CAM therapies they (1) knew enough about to discuss with patients, (2) considered part of "legitimate medical practice," and (3) "personally administered" to patients, or "referred patients to someone else" to administer. The response rate was 47%. RESULTS: On average, the respondents reported knowing enough to discuss 10 of the therapies with patients, considered 9 to be part of legitimate medical practice, and had referred patients to someone else for 8 of the 22 therapies. Correlates of use and/or referral included sex, age, belief in the legitimacy of the therapies, and self-reported knowledge. CONCLUSIONS: These results provide potentially important preliminary data regarding rheumatologists' responses to dramatic increases in the use of CAM therapies among their patients. PMID- 11926851 TI - Pharmaceutical manufacturer assistance programs. AB - BACKGROUND AND OBJECTIVE: While there have been many public discussions concerning Medicare covering outpatient prescription medications, currently, Medicare does not pay for most outpatient prescriptions. Meanwhile, costs associated with prescription medications as well as the number of underinsured patients are rapidly increasing. Many pharmaceutical companies offer assistance programs for patients who require medications but have inadequate financial resources to obtain them. Because patient access to required prescriptions is critical and clinician involvement is necessary for patient enrollment, the purpose of this article is to facilitate awareness of pharmaceutical companies' assistance programs, the availability of such programs, and their enrollment process. METHODS: English-language articles from MEDLINE (1963-2000) and Internet Web pages describing medication assistance programs were reviewed. Data obtained from pharmaceutical companies' medication assistance programs were also included. In addition to general information concerning medication assistance programs, all studies found in the literature search describing the clinical and financial impact of using these programs and data obtained from contacting several medication assistance programs were selected. To determine if an assistance program was available for a medication listed as one of the top 200 medications prescribed in the United States, we contacted the pharmaceutical company that manufactures each medication. RESULTS: Approximately 53% of the top 200 prescribed medications in 1999 were offered through assistance programs to indigent patients. Physician office personnel can obtain medications for eligible patients by completing the enrollment process. CONCLUSION: Pharmaceutical companies' medication assistance programs can be used to reduce individual patient drug expenditures and improve patient outcomes by increasing medication access to those in need. PMID- 11926850 TI - Association between thyroid dysfunction and total cholesterol level in an older biracial population: the health, aging and body composition study. AB - BACKGROUND: Thyroid dysfunction increases with age. Less is known about the prevalence of thyroid disease in older black adults and whether an association between thyroid function and serum cholesterol level exists, as in older white adults. METHODS: A cross-sectional study of 2799 well functioning white and black participants, aged 70 to 79 years, were recruited for a population-based study. Participants underwent thyrotropin, free thyroxine, and total cholesterol testing; a medical history; and physical measurements. RESULTS: Among the entire cohort, 94% were euthyroid based on biochemical testing results. Approximately 10% were taking thyroid hormones. Subclinical hypothyroidism was the most prevalent disorder (3.1% of all participants not taking thyroid hormones), but black men and women had lower rates of this condition than white men and women. After excluding those taking thyroid or lipid medication and adjusting for potential confounders, an elevated thyrotropin level (>5.5 mIU/mL) was associated with a 9 mg/dL (0.23 mmol/L) higher cholesterol level, and a suppressed thyrotropin level (<0.35 mIU/mL) was associated with a 19 mg/dL (0.49 mmol/L) lower cholesterol level. CONCLUSION: Healthy community-dwelling older black adults have a lower prevalence of thyroid dysfunction compared with older white adults, but the association between increased thyrotropin and increased cholesterol levels is similar in both races. PMID- 11926852 TI - Chlamydia pneumoniae seropositivity and systemic and renovascular atherosclerotic disease. AB - BACKGROUND: Patients with hypertension may be vulnerable to vascular Chlamydia pneumoniae and/or cytomegalovirus (CMV) infection because of increased expression of adhesion molecules. OBJECTIVE: To determine whether C pneumoniae or CMV is associated with the presence of atherosclerotic lesions in hypertensive patients. METHODS: Ninety-six angiographic studies on 100 consecutive patients with of clinical signs or symptoms suggestive of renovascular hypertension were reviewed for the presence or absence of atherosclerotic lesions at the level of the renal arteries as well and abdominal aorta. Also, the presence of a hemodynamically notable renal artery stenosis and antibodies to C pneumoniae (IgG and IgA) and CMV (IgG and IgM) was determined, and all classic risk factors were recorded. RESULTS: Atherosclerotic lesions were documented in 67 patients (70%), and in 49 patients (51%) such lesions were present at the level of the renal artery. In the univariate analysis, significant associations between IgG (odds ratio, 3.8; 95% confidence interval, 1.2-11.7; P =.02) as well as IgA (odds ratio, 2.6; 95% confidence interval, 1.1-6.7; P =.03) antibodies to C pneumoniae and the presence of atherosclerosis were found for both the aorta and the renal arteries. Seroprevalence (IgG) to C pneumoniae in the 23 patients with a hemodynamically notable renal artery stenosis was 100% and differed (P =.01) from those without a notable renal artery stenosis (78%). In the multivariate analysis, IgG seropositivity to C pneumoniae was significantly associated with atherosclerosis (odds ratio, 6.0; 95% confidence interval, 1.33-27.5; P =.02), and age. There was no association between CMV seropositivity and atherosclerosis. CONCLUSION: The presence of antibodies to C pneumoniae was significantly associated with atherosclerosis and renovascular disease in hypertensive patients in whom a renal artery stenosis was strongly suspected. PMID- 11926853 TI - Can a Back Pain E-mail Discussion Group improve health status and lower health care costs?: A randomized study. AB - BACKGROUND: Given the high health care utilization, limited evidence for the effectiveness of back pain interventions, and the proliferation of e-mail health discussion groups, this study seeks to determine if the Internet can be used to improve health status and health care utilization for people with chronic back pain. METHODS: Randomized controlled trial. Participants included 580 people from 49 states with chronic back pain having at least 1 outpatient visit in the past year, no "red-flag" symptoms, and access to e-mail. Major exclusion criteria included continuous back pain for more than 90 days causing major activity intolerance and/or receiving disability payments. INTERVENTION: Closed, moderated, e-mail discussion group. Participants also received a book and videotape about back pain. Controls received a subscription to a non-health related magazine of their choice. MAIN OUTCOME MEASURES: Pain, disability, role function, health distress, and health care utilization. RESULTS: At 1-year treatment, subjects compared with controls demonstrated improvements in pain (P =.045), disability (P =.02), role function (P =.007), and health distress (P =.001). Physician visits for the past 6 months declined by 1.5 visits for the treatment group and by 0.65 visits for the control group (P =.07). Mean hospital days declined nearly 0.20 days for the treated group vs and increased 0.04 days for the control group (P =.24). CONCLUSIONS: An e-mail discussion group can positively affect health status and possibly health care utilization. It may have a place in the treatment of chronic recurrent back pain. PMID- 11926854 TI - A community-wide survey of physician practices and attitudes toward cholesterol management in patients with recent acute myocardial infarction. AB - BACKGROUND: Physicians' current attitudes and practices toward the management of high cholesterol levels in patients with recent acute myocardial infarction are not well defined. OBJECTIVE: To examine threshold levels of serum cholesterol and other factors that influence physicians' decision to prescribe lipid-lowering drugs and initiate dietary therapy in patients with recent acute myocardial infarction. METHODS: Community-wide questionnaire survey of general internists, cardiologists, and family physicians practicing in the Worcester, Mass, metropolitan area. RESULTS: Among the 257 responding physicians, lipid-lowering drug therapy was more likely to be initiated in younger patients at lower total serum and low-density lipoprotein (LDL) cholesterol levels than in older patients (P =.03). Younger physicians were more likely to initiate dietary and lipid lowering drug therapy at lower total and LDL cholesterol levels than their older counterparts. Younger physicians also considered LDL cholesterol level the most important factor in initiating lipid-lowering drug therapy in contrast to older physicians who favored total cholesterol level (P =.001). General practice physicians were more likely to initiate dietary therapy at lower total cholesterol levels, but tended to initiate lipid-lowering drug therapy at higher total and LDL cholesterol levels compared with internists and cardiologists. Physicians reported that the most important factors that interfere with patients' use of lipid-lowering medication were concerns about medication costs, issues related to polypharmacy, and failure to recognize the importance of lipid lowering drugs. Several physician-associated factors, including perceived importance of other cardiac drugs and provider responsibility, were associated with the nonuse of lipid-lowering medications. CONCLUSION: Educational and practice-based efforts remain necessary to remove potential barriers to the implementation of effective long-term cholesterol management in patients with recent acute myocardial infarction. PMID- 11926856 TI - Smoking and alanine aminotransferase levels in hepatitis C virus infection: implications for prevention of hepatitis C virus progression. AB - BACKGROUND: Alcohol consumption is a well-known risk factor for elevated ALT levels, but the role of cigarette smoking is unclear. METHODS: We collected a cross-sectional sample of 6095 inhabitants 35 years or older in a community with hyperendemic hepatitis B and C virus infections. We assayed levels of serum alanine aminotransferase (ALT), hepatitis B surface antigen (HBsAg), and anti hepatitis C virus antibody (anti-HCV). Multivariate logistic regression was performed to determine the factors for elevated ALT levels (> or =40 U/L) among people with different hepatitis infection statuses. RESULTS: Prevalence of elevated ALT levels in individuals who were seronegative for both infections or seropositive for HBsAg or anti-HCV was 3.9%, 11.1%, and 30.8%, respectively. Subjects with elevated ALT levels were more likely to be seropositive for anti HCV, male, and seropositive for HBsAg; to drink alcohol; to smoke; and to have undergone blood transfusion (P<.05). An association was found between elevated ALT levels and the consumption of cigarettes and alcohol among anti-HCV seropositive subjects. In multivariate logistic analyses, alcohol consumption (odds ratio [OR], 2.2; 95% confidence interval [CI], 1.2-4.1) and smoking (OR, 1.8; 95% CI, 1.1-2.7) were significantly associated with elevated ALT levels among anti-HCV-seropositive subjects, but no such association was found among HBsAg-seropositive subjects. The odds of elevated ALT levels were 7 times higher (95% CI, 2.7-18.8) for the anti-HCV-seropositive patients who smoked 1 or more packs of cigarettes per day and frequently drank alcohol than for those who did not. CONCLUSIONS: Smoking and alcohol consumption are independently associated with elevated ALT levels among anti-HCV-seropositive individuals but not among HBsAg-seropositive individuals. Patients who are seropositive for anti-HCV are strongly advised not to smoke and drink alcohol to reduce the possible risk for aggravating the liver dysfunction. PMID- 11926855 TI - Risk of hepatitis C virus transmission from an infected gynecologist to patients: results of a 7-year retrospective investigation. AB - BACKGROUND: Currently, it is not known how often hepatitis C virus (HCV) is transmitted from infected health care workers to patients during medical care. In the present investigation, we tried to determine the rate of provider-to-patient transmission of HCV among former patients of an HCV-positive gynecologist after it was proven that he infected one of his patients with HCV during a cesarean section. METHODS: All 2907 women who had been operated on by the HCV-positive gynecologist between July 1993 and March 2000 were notified about potential exposure and were offered free counseling and testing. The crucial differentiation between HCV transmissions caused by the gynecologist and infections contracted from other sources was achieved by epidemiological investigations, nucleotide sequencing, and phylogenetic analysis. RESULTS: Of the 2907 women affected, 78.6% could be screened for markers of HCV infection. Seven of these former patients were found to have HCV. Phylogenetic analysis of HCV sequences from the gynecologist and the women did not indicate that the virus strains were linked. Therefore, no further iatrogenic HCV infections caused by the gynecologist could be detected. The resulting overall HCV transmission rate was 0.04% (1 per 2286; 95% confidence interval, 0.008%-0.25%). CONCLUSION: To our knowledge, this is the largest retrospective investigation of the risk of provider-to-patient transmission of HCV conducted so far. Our findings support the notion that such transmissions are relatively rare events and might provide a basis for future recommendations on the management of HCV-infected health care workers. PMID- 11926857 TI - Opposite associations of carbohydrate-deficient transferrin and gamma glutamyltransferase with prevalent coronary heart disease. AB - BACKGROUND: Moderate alcohol consumption may reduce the risk of coronary heart disease (CHD), but excessive alcohol consumption is probably harmful to the heart. We analyzed the association of 2 commonly used markers of alcohol consumption-carbohydrate-deficient transferrin (CDT) and gamma glutamyltransferase (GGT)-and self-reported alcohol consumption with prevalent CHD. METHODS: The study included a random sample of 3666 Finnish men aged 25 to 74 years who participated in a risk factor survey in 1997. The cross-sectional association of CDT, GGT, and self-reported drinking with CHD was analyzed by means of logistic regression models. RESULTS: The CDT level was inversely and GGT level positively associated with CHD risk. The odds ratios (adjusted for age, smoking, total and high-density lipoprotein cholesterol levels, systolic blood pressure, and body mass index) of CHD among men in the fourth quartiles of CDT and GGT, as compared with the first quartiles, were 0.69 and 1.76, respectively. In a composite risk assessment, men with normal CDT levels (< or =20 U/L) and elevated GGT levels (>80 U/L) had nearly 8-fold adjusted risk of CHD as compared with the men with normal GGT levels and elevated CDT levels. Self-reported alcohol consumption had an inverse association with CHD risk, which disappeared after adjustment for the other risk factors. CONCLUSIONS: Levels of CDT and GGT may be indicators of factors behind the curvilinear association between alcohol consumption and CHD risk. The CDT level seems to be related to beneficial biological changes and GGT level with the changes that are detrimental to the cardiovascular system. The inverse association of CDT level with CHD risk will be examined further in a forthcoming prospective study. PMID- 11926858 TI - Lack of penicillin resensitization in patients with a history of penicillin allergy after receiving repeated penicillin courses. AB - BACKGROUND: Up to 10% of the population reports an allergy to penicillin, yet more than 80% of these individuals lack penicillin-specific IgE antibodies. A negative result on a penicillin skin test is highly accurate in identifying who can safely receive the antibiotic at the time of testing. However, its negative predictive value for future courses is unknown because it is uncertain whether patients with a history of penicillin allergy are at risk of becoming resensitized. OBJECTIVE: To determine the rate of penicillin resensitization in adult patients with a history of penicillin allergy after they are challenged with repeated courses of oral penicillin. METHODS: Adult patients with a history of penicillin allergy consistent with an IgE-mediated mechanism were recruited and underwent penicillin skin testing. Those with negative skin test results were challenged with 3 successive 10-day courses of penicillin V potassium (250 mg by mouth 3 times a day), providing their penicillin skin test results remained negative prior to each course. Patients with positive skin test results were not challenged. RESULTS: Of 53 patients with initially negative skin test results, 46 completed the protocol, and each tolerated all 3 courses of penicillin with negative skin test results throughout. No patients had a converted skin test result from negative to a positive, yielding a resensitization rate of 0% (upper 95% confidence interval, 2.1%). CONCLUSIONS: Adult patients with a history of penicillin allergy are not at increased risk of resensitization after receiving 3 courses of oral penicillin. Because a negative penicillin skin test result is predictive for subsequent oral administrations beyond the time of testing, adult patients with a history of penicillin allergy can be skin tested electively, which may avoid unnecessary treatment with alternate broad-spectrum antibiotics. PMID- 11926859 TI - Patient and hospital characteristics associated with recommended processes of care for elderly patients hospitalized with pneumonia: results from the medicare quality indicator system pneumonia module. AB - BACKGROUND: Unexplained wide variability exists in the performance of key initial processes of care associated with improved survival of elderly patients (those > or =65 years) hospitalized with pneumonia. The objective of this study was to assess which patient and hospital characteristics are associated with performance of these key initial processes of care for hospitalized elderly patients with pneumonia. METHODS: A retrospective cohort analysis was performed using data from the Medicare Quality Indicator System Pneumonia Module for 14 069 patients 65 years or older hospitalized with pneumonia throughout the United States. Associations were calculated using multivariate logistic regression analysis between specific patient and hospital characteristics and 2 processes of care associated with improved 30-day survival: administration of antibiotics within 8 hours of hospital arrival and blood culture collection within 24 hours of arrival. RESULTS: Timely antibiotic administration was negatively associated with nonwhite race (African American: odds ratio [OR], 0.71; 95% confidence interval [CI], 0.60-0.85; and other racial minorities: OR, 0.79; 95% CI, 0.68-0.92), major hospital teaching status (OR, 0.79; 95% CI, 0.67-0.93), and larger hospital size (> or =250 beds vs. <100 beds: OR, 0.68; 95% CI, 0.59-0.80). Timely blood culture collection was positively associated with larger hospital size (OR, 1.61; 95% CI, 1.39-1.87). Performance of both processes of care were positively associated with registered nurse-bed ratios of 1.25 or higher (for antibiotic administration: OR, 1.23; 95% CI, 1.10-1.38; and for blood culture collection: OR, 1.43; 95% CI, 1.26 1.61) and fever (for antibiotic administration: OR, 1.35; 95% CI, 1.23-1.49; and for blood culture collection: OR, 3.07; 95% CI, 2.81-3.34) and were negatively associated with hospital location in the South (for antibiotic administration: OR, 0.77; 95% CI, 0.69-0.86; and for blood culture collection: OR, 0.85; 95% CI, 0.77-0.93). CONCLUSIONS: Minority race, fever, nurse-bed ratio, hospital size and teaching status, and southern location are among the major patient and hospital characteristics associated, either negatively or positively, with the timeliness of performance of initial antibiotic administration and blood culture collection for patients hospitalized with pneumonia. Because performance of these processes of care is associated with improved likelihood of survival, medical providers should seek to eliminate the variations in care associated with these patient and hospital characteristics. In addition, the impact of nurse staffing changes on performance of key time-sensitive processes of care should be weighed carefully. PMID- 11926860 TI - Drug-induced lupus erythematosus by amiodarone. AB - We report a case with typical clinical features of drug-induced lupus erythematosus, together with verrucous endocarditis and pleuropericardial effusion, due to amiodarone treatment. After cessation of amiodarone treatment, the patient recovered completely. PMID- 11926861 TI - Epstein-Barr viral load assessment in immunocompetent patients with fulminant infectious mononucleosis. AB - We describe 2 immunocompetent adolescents with fulminant infectious mononucleosis and virus-associated hemophagocytosis. A new quantitative polymerase chain reaction revealed high serum Epstein-Barr virus DNA levels in these patients. One patient died with an increasing viral load not responding to corticosteroids followed by antiviral and intensified immunomodulatory treatment. The other patient received corticosteroids and acyclovir at diagnosis; her rapid recovery was heralded by a steep decline of viral load. We propose monitoring the clinical course of fulminant infectious mononucleosis in immunocompetent patients by Epstein-Barr virus DNA quantification and prompt corticosteroid and antiviral therapy when viral load is high. PMID- 11926862 TI - Comments, opinions, and brief case reports: desmopressin test in occult eutopic corticotropin microadenoma. PMID- 11926863 TI - Is there proof that captopril causes proteinuria? PMID- 11926865 TI - Is erythema multiforme associated with bupropion use? PMID- 11926866 TI - Pneumonia treatment process and quality. PMID- 11926867 TI - Dilatory standards are below the level of care. PMID- 11926869 TI - Emergency department treatment of migraine headaches. PMID- 11926871 TI - A piece of my mind. Exclusive interview with Helicobacter pylori. PMID- 11926873 TI - Researchers explore new anti-HIV agents. PMID- 11926874 TI - Scientists find some genes a bad omen for anti-HIV drug. PMID- 11926875 TI - Clinical evidence for critical cardiac care. PMID- 11926876 TI - From the Food and Drug Administration. PMID- 11926878 TI - Drug therapy for patients with type 2 diabetes. PMID- 11926879 TI - Drug therapy for patients with type 2 diabetes. PMID- 11926881 TI - Treatment decisions for type 2 diabetes. PMID- 11926884 TI - The costs of making practice more cost-effective. PMID- 11926883 TI - The costs of making practice more cost-effective. PMID- 11926885 TI - The costs of making practice more cost-effective. PMID- 11926886 TI - The costs of making practice more cost-effective. PMID- 11926888 TI - Poliomyelitis and Parkinson disease. PMID- 11926889 TI - Dopamine transporter brain imaging to assess the effects of pramipexole vs levodopa on Parkinson disease progression. AB - CONTEXT: Pramipexole and levodopa are effective medications to treat motor symptoms of early Parkinson disease (PD). In vitro and animal studies suggest that pramipexole may protect and that levodopa may either protect or damage dopamine neurons. Neuroimaging offers the potential of an objective biomarker of dopamine neuron degeneration in PD patients. OBJECTIVE: To compare rates of dopamine neuron degeneration after initial treatment with pramipexole or levodopa in early PD by means of dopamine transporter imaging using single-photon emission computed tomography (SPECT) with 2beta-carboxymethoxy-3beta(4-iodophenyl)tropane (beta-CIT) labeled with iodine 123. DESIGN: Substudy of a parallel-group, double blind randomized clinical trial. SETTING AND PATIENTS: Eighty-two patients with early PD who were recruited at 17 clinical sites in the United States and Canada and required dopaminergic therapy to treat emerging disability, enrolled between November 1996 and August 1997. INTERVENTIONS: Patients were randomly assigned to receive pramipexole, 0.5 mg 3 times per day with levodopa placebo (n = 42), or carbidopa/levodopa, 25/100 mg 3 times per day with pramipexole placebo (n = 40). For patients with residual disability, the dosage was escalated during the first 10 weeks, and subsequently, open-label levodopa could be added. After 24 months of follow-up, the dosage of study drug could be further modified. MAIN OUTCOME MEASURES: The primary outcome variable was the percentage change from baseline in striatal [(123)I]beta-CIT uptake after 46 months. The percentage changes and absolute changes in striatal, putamen, and caudate [(123)I]beta-CIT uptake after 22 and 34 months were also assessed. Clinical severity of PD was assessed using the Unified Parkinson Disease Rating Scale (UPDRS) 12 hours off anti-PD medications. RESULTS: Sequential SPECT imaging showed a decline in mean (SD) [(123)I]beta-CIT striatal uptake from baseline of 10.3% (9.8%) at 22 months, 15.3% (12.8%) at 34 months, and 20.7% (14.4%) at 46 months-approximately 5.2% per year. The mean (SD) percentage loss in striatal [(123)I]beta-CIT uptake from baseline was significantly reduced in the pramipexole group compared with the levodopa group: 7.1% (9.0%) vs 13.5% (9.6%) at 22 months (P =.004); 10.9% (11.8%) vs 19.6% (12.4%) at 34 months (P =.009); and 16.0% (13.3%) vs 25.5% (14.1%) at 46 months (P =.01). The percentage loss from baseline in striatal [(123)I]beta-CIT uptake was correlated with the change from baseline in UPDRS at the 46-month evaluation (r = - 0.40; P =.001). CONCLUSIONS: Patients initially treated with pramipexole demonstrated a reduction in loss of striatal [(123)I]beta-CIT uptake, a marker of dopamine neuron degeneration, compared with those initially treated with levodopa, during a 46-month period. These imaging data highlight the need to further compare imaging and clinical end points of PD progression in long-term studies. PMID- 11926890 TI - alpha-Methylacyl coenzyme A racemase as a tissue biomarker for prostate cancer. AB - CONTEXT: Molecular profiling of prostate cancer has led to the identification of candidate biomarkers and regulatory genes. Discoveries from these genome-scale approaches may have applicability in the analysis of diagnostic prostate specimens. OBJECTIVES: To determine the expression and clinical utility of alpha methylacyl coenzyme A racemase (AMACR), a gene identified as being overexpressed in prostate cancer by global profiling strategies. DESIGN: Four gene expression data sets from independent DNA microarray analyses were examined to identify genes expressed in prostate cancer (n = 128 specimens). A lead candidate gene, AMACR, was validated at the transcript level by reverse transcriptase polymerase chain reaction (RT-PCR) and at the protein level by immunoblot and immunohistochemical analysis. AMACR levels were examined using prostate cancer tissue microarrays in 342 samples representing different stages of prostate cancer progression. Protein expression was characterized as negative (score = 1), weak (2), moderate (3), or strong (4). Clinical utility of AMACR was evaluated using 94 prostate needle biopsy specimens. MAIN OUTCOME MEASURES: Messenger RNA transcript and protein levels of AMACR; sensitivity and specificity of AMACR as a tissue biomarker for prostate cancer in needle biopsy specimens. RESULTS: Three of 4 independent DNA microarray analyses (n = 128 specimens) revealed significant overexpression of AMACR in prostate cancer (P<.001). AMACR up-regulation in prostate cancer was confirmed by both RT-PCR and immunoblot analysis. Immunohistochemical analysis demonstrated an increased expression of AMACR in malignant prostate epithelia relative to benign epithelia. Tissue microarrays to assess AMACR expression in specimens consisting of benign prostate (n = 108 samples), atrophic prostate (n = 26), prostatic intraepithelial neoplasia (n = 75), localized prostate cancer (n = 116), and metastatic prostate cancer (n = 17) demonstrated mean AMACR protein staining intensity of 1.31 (95% confidence interval, 1.23-1.40), 2.33 (95% CI, 2.13-2.52), 2.67 (95% CI, 2.52-2.81), 3.20 (95% CI, 3.10-3.28), and 2.50 (95% CI, 2.20-2.80), respectively (P<.001). Pairwise comparisons demonstrated significant differences in staining intensity between clinically localized prostate cancer compared with benign prostate tissue, with mean expression scores of 3.2 and 1.3, respectively (mean difference, 1.9; 95% CI, 1.7-2.1; P<.001). Using moderate or strong staining intensity as positive (score = 3 or 4), evaluation of AMACR protein expression in 94 prostate needle biopsy specimens demonstrated 97% sensitivity and 100% specificity for detecting prostate cancer. CONCLUSIONS: AMACR was shown to be overexpressed in prostate cancer using independent experimental methods and prostate cancer specimens. AMACR may be useful in the interpretation of prostate needle biopsy specimens that are diagnostically challenging. PMID- 11926891 TI - Osteopontin as a potential diagnostic biomarker for ovarian cancer. AB - CONTEXT: Development of new biomarkers for ovarian cancer is needed for early detection and disease monitoring. Analyses involving complementary DNA (cDNA) microarray data can be used to identify up-regulated genes in cancer cells, whose products may then be further validated as potential biomarkers. OBJECTIVE: To describe validation studies of an up-regulated gene known as osteopontin, previously identified using a cDNA microarray system. DESIGN, SETTING, AND PARTICIPANTS: Experimental and cross-sectional studies were conducted involving ovarian cancer and healthy human ovarian surface epithelial cell lines and cultures, archival paraffin-embedded ovarian tissue collected between June 1992 and June 2001, and fresh tissue and preoperative plasma from 144 patients evaluated for a pelvic mass between June 1992 and June 2001 in gynecologic oncology services at 2 US academic institutions. Plasma samples from 107 women selected from an epidemiologic study of ovarian cancer initiated between May 1992 and March 1997 were used as healthy controls. MAIN OUTCOME MEASURES: Relative messenger RNA expression in cancer cells and fresh ovarian tissue, measured by real-time polymerase chain reaction as 2(-DeltaDeltaCT)(a quantitative value representing the amount of osteopontin expression); osteopontin production, localized and scored in ovarian healthy and tumor tissue with immunohistochemical studies; and amount of osteopontin in patient vs control plasma, measured using an enzyme-linked immunoassay. RESULTS: The geometric mean for 2(-DeltaDeltaCT)for osteopontin expression in 5 healthy ovarian epithelial cell cultures was 4.1 compared with 270.4 in 14 ovarian cancer cell lines (P =.03). The geometric mean 2(-DeltaDeltaCT)for osteopontin expression in tissue from 2 healthy ovarian epithelial samples was 9.0 compared with 164.0 in 27 microdissected ovarian tumor tissue samples (P =.06). Immunolocalization of osteopontin showed that tissue samples from 61 patients with invasive ovarian cancer and 29 patients with borderline ovarian tumors expressed higher levels of osteopontin than tissue samples from 6 patients with benign tumors and samples of healthy ovarian epithelium from 3 patients (P =.03). Osteopontin levels in plasma were significantly higher (P<.001) in 51 patients with epithelial ovarian cancer (486.5 ng/mL) compared with those of 107 healthy controls (147.1 ng/mL), 46 patients with benign ovarian disease (254.4 ng/mL), and 47 patients with other gynecologic cancers (260.9 ng/mL). CONCLUSIONS: Our findings provide evidence for an association between levels of a biomarker, osteopontin, and ovarian cancer and suggest that future research assessing its clinical usefulness would be worthwhile. PMID- 11926892 TI - Diuretic therapy, the alpha-adducin gene variant, and the risk of myocardial infarction or stroke in persons with treated hypertension. AB - CONTEXT: A genetic variant in alpha-adducin has been associated with renal sodium reabsorption and salt-sensitive hypertension. Whether this genetic variant modifies the effect of diuretic therapy on the incidence of myocardial infarction (MI) and stroke is unknown. OBJECTIVES: To estimate the interaction between alpha adducin and diuretic therapy on the risk of MI or stroke. Specifically, we hypothesized that in participants with treated hypertension, the risk of MI or stroke associated with diuretic use would be lower in carriers of the adducin variant than in carriers of the adducin wild-type genotype. DESIGN, SETTING, AND PARTICIPANTS: Population-based case-control study of patients enrolled in a health maintenance organization, treated pharmacologically for hypertension, and genotyped as homozygous carriers of the adducin wild-type genotype or carriers of 1 or 2 copies of the Trp460 variant allele. Cases had a first nonfatal MI (n = 206) or stroke (n = 117) between January 1995 and December 1998. Controls (n = 715) were a stratified random sample of pharmacologically treated hypertensive patients who were matched to MI cases by age, sex, and calendar year. MAIN OUTCOME MEASURE: Risk of the combined outcome of first nonfatal MI or stroke. RESULTS: The adducin variant was present in more than one third of the participants. Among the 653 carriers of the adducin wild-type genotype, diuretic therapy was not associated with the risk of MI or stroke (odds ratio [OR], 1.09; 95% confidence interval [CI], 0.78-1.52). Among the 385 carriers of the adducin variant allele, diuretic therapy was associated with a lower risk of the combined outcome of MI and stroke than other antihypertensive therapies (OR, 0.49; 95% CI, 0.32-0.77). The OR in carriers of the adducin variant was less than half of the OR in carriers of the wild-type genotype (P =.005). The case-control synergy index (SI) was 0.45 (95% CI, 0.26-0.79) for the combined outcome of MI and stroke. The point estimates of the diuretic-adducin interaction were similar in separate analyses of MI (SI, 0.41; 95% CI, 0.21-0.80) and stroke (SI, 0.53; 95% CI, 0.24-1.19). The diuretic-adducin interaction was not confounded by traditional cardiovascular risk factors, was specific to diuretic therapy but not present for other major antihypertensive drug classes, and did not differ substantially between subgroups defined by age, sex, race, diabetes, and history of cardiovascular disease. CONCLUSIONS: In carriers of the adducin variant, diuretic therapy was associated with a lower risk of combined MI or stroke than other antihypertensive therapies. If these findings are confirmed in other studies, this large subgroup of the hypertensive population may be especially likely to benefit from low-dose diuretic therapy. PMID- 11926893 TI - Association between CYP2C9 genetic variants and anticoagulation-related outcomes during warfarin therapy. AB - CONTEXT: Warfarin is a commonly used anticoagulant that requires careful clinical management to balance the risks of overanticoagulation and bleeding with those of underanticoagulation and clotting. The principal enzyme involved in warfarin metabolism is CYP2C9, and 2 relatively common variant forms with reduced activity have been identified, CYP2C9*2 and CYP2C9*3. Patients with these genetic variants have been shown to require lower maintenance doses of warfarin, but a direct association between CYP2C9 genotype and anticoagulation status or bleeding risk has not been established. OBJECTIVE: To determine if CYP2C9*2 and CYP2C9*3 variants are associated with overanticoagulation and bleeding events during warfarin therapy. DESIGN AND SETTING: Retrospective cohort study conducted at 2 anticoagulation clinics based in Seattle, Wash. PARTICIPANTS: Two hundred patients receiving long-term warfarin therapy for various indications during April 3, 1990, to May 31, 2001. Only patients with a complete history of warfarin exposure were included. MAIN OUTCOME MEASURES: Anticoagulation status, measured by time to therapeutic international normalized ratio (INR), rate of above-range INRs, and time to stable warfarin dosing; and time to serious or life-threatening bleeding events. RESULTS: Among 185 patients with analyzable data, 58 (31.4%) had at least 1 variant CYP2C9 allele and 127 (68.6%) had the wild-type (*1/*1) genotype. Mean maintenance dose varied significantly among the 6 genotype groups (*1/*1 [n = 127], *1/*2 [n = 28], *1/*3 [n = 18], *2/*2 [n = 4], *2/*3 [n = 3], *3/*3 [n = 5]) (by Kruskall-Wallis test, chi(2)(5) = 37.348; P<.001). Compared with patients with the wild-type genotype, patients with at least 1 variant allele had an increased risk of above-range INRs (hazard ratio [HR], 1.40; 95% confidence interval [CI], 1.03-1.90). The variant group also required more time to achieve stable dosing (HR, 0.65; 95% CI, 0.45-0.94), with a median difference of 95 days (P =.004). In addition, although numbers were small for some genotypes, representing potentially unstable estimates, patients with a variant genotype had a significantly increased risk of a serious or life-threatening bleeding event (HR, 2.39; 95% CI, 1.18-4.86). CONCLUSIONS: The results of our study suggest that the CYP2C9*2 and CYP2C9*3 polymorphisms are associated with an increased risk of overanticoagulation and of bleeding events among patients in a warfarin anticoagulation clinic setting, although small numbers in some cases would suggest the need for caution in interpretation. Screening for CYP2C9 variants may allow clinicians to develop dosing protocols and surveillance techniques to reduce the risk of adverse drug reactions in patients receiving warfarin. PMID- 11926894 TI - Nontypeable Haemophilus influenzae in carriage and disease: a difference in IgA1 protease activity levels. AB - CONTEXT: Nontypeable Haemophilus influenzae strains form part of the normal flora of the human upper respiratory tract but are also implicated in a wide range of diseases. Infections caused by nontypeable H influenzae are major health and socioeconomic burdens. No single bacterial trait has been associated with disease as opposed to colonization. OBJECTIVES: To compare IgA1 protease activity in nontypeable H influenzae strains isolated from patients with symptomatic Haemophilus infection (sputum, cerebrospinal fluid, blood, or normally sterile tissue) vs strains from throat swabs of asymptomatic carriers and to compare iga gene carriage and variability in nontypeable H influenzae strains. DESIGN AND SETTING: Retrospective study of 63 strains (44 clinical and 19 carriage) collected between 1991 and 2000 and maintained at the Public Health Laboratory, Gwynedd General Hospital, Bangor, Wales. MAIN OUTCOME MEASURES: Levels of IgA1 protease activity produced by carriage strains and clinical isolates from symptomatic patients; the determination of the size and sequence of a variable region of the iga gene. RESULTS: Bacterial IgA1 protease activity was significantly higher (P<.001) in strains isolated from sputum, blood, cerebrospinal fluid, or normally sterile tissue of symptomatic individuals (median, 155 mU; interquartile range [IQR], 80-220 mU; mean, 169 mU; 95% confidence interval [CI], 126-211 mU) than in those isolated from throat swabs of asymptomatic carriers (median, 30 mU; IQR, 15-90 mU; mean, 56 mU; 95% CI, 26-86 mU; assayed on secretory IgA). The iga gene was detected in 97% of all strains examined. Variations in the sizes and sequences of part of the iga genes were also apparent. This variable region encodes a polypeptide linker connecting the protease domain to the beta-core autotranslocator, a porelike structure required for secretion of the protease. CONCLUSIONS: These findings reveal the importance of iga gene variability and expression levels in the establishment of disease phenotype. They identify nontypeable H influenzae IgA1 protease as a virulence factor and as a potential target for the development of new strategies to fight these important pathogens. PMID- 11926895 TI - Effect of leptin on arterial thrombosis following vascular injury in mice. AB - CONTEXT: Complications of atherosclerosis are the leading cause of morbidity and mortality in industrialized societies. Obesity has emerged as an independent risk factor for complications of atherosclerotic vascular disease. Leptin, a hormone produced by the adipocyte, increases with obesity and appears to modulate energy balance and food intake. In addition, other actions of leptin have been proposed, including an in vitro effect on platelet aggregation. Thus, the elevated plasma leptin levels in obese individuals may promote vascular thrombosis. OBJECTIVE: To test the hypothesis that leptin contributes to in vivo thrombosis via the leptin receptor. DESIGN AND MATERIALS: Between September 2000 and September 2001, a vascular thrombosis model was used to test male 10- to 12-week-old mice completely deficient in leptin or the leptin receptor and mice with platelet leptin-receptor deficiency. MAIN OUTCOME MEASURE: Time to formation of an occlusive thrombus in the common carotid artery following experimentally induced endothelial injury. RESULTS: Following onset of vascular injury, wild-type mice (n = 8) formed occlusive thrombosis in a mean (SD) of 42.2 (4.6) minutes, whereas leptin-deficient (n = 5) and leptin receptor-deficient mice (n = 7) formed occlusive thrombosis in 75.2 (10.1) and 68.6 (10.3) minutes, respectively (leptin deficient vs wild-type mice, P =.008; leptin-receptor-deficient vs wild-type, P =.03). When recombinant murine leptin was administered to leptin-deficient mice (n = 4), the time to occlusion was reduced to 41.8 (6.6) minutes (P =.035 vs vehicle control). Following bone marrow transplantation from leptin receptor deficient (donor) mice to wild-type (recipient) mice, the time to occlusion was prolonged from 22.3 (2.8) minutes in wild-type mice receiving wild-type marrow (n = 3) to 56.8 (5.0) minutes in wild-type mice receiving leptin receptor-deficient bone marrow (n = 5) (P =.003). CONCLUSIONS: Leptin contributes to arterial thrombosis following vascular injury in vivo and these prothrombotic effects appear to be mediated through the platelet leptin receptor. PMID- 11926896 TI - Mucosal biofilm formation on middle-ear mucosa in the chinchilla model of otitis media. AB - CONTEXT: Chronic otitis media with effusion (OME) has long been considered to be a sterile inflammatory process. The previous application of molecular diagnostic technologies to OME suggests that viable bacteria are present in complex communities known as mucosal biofilms; however, direct imaging evidence of mucosal biofilms associated with OM is lacking. OBJECTIVE: To determine whether biofilm formation occurs in middle-ear mucosa in an experimental model of otitis media. DESIGN AND MATERIALS: A total of 48 research-grade, young adult chinchillas weighing 500 g were used for 2 series of animal experiments: one to obtain specimens for scanning electron microscopy and the other to obtain specimens for confocal laser scanning microscopy using vital dyes. In each series, 21 animals were bilaterally injected with viable Haemophilus influenzae bacteria and 1 was inoculated to account for expected mortality. Three served as negative controls. Effusions and mucosal specimens were collected from 2 infected animals that were euthanized at 3, 6, 12, and 24 hours and at days 2, 4, 5, 10, 16, and 22 after inoculation. MAIN OUTCOME MEASURES: Images were analyzed for biofilm morphology, including presence of microcolony formation and for presence of bacteria on tissue surfaces. RESULTS: Scanning electron microscopy demonstrated that biofilm formation was evident in all specimens from animals beginning 1 day after infection and was present through 21 days. Confocal laser scanning microscopy indicated that bacteria within the biofilms are viable. CONCLUSION: These preliminary findings provide evidence that mucosal biofilms form in an experimental model of otitis media and suggest that biofilm formation may be an important factor in the pathogenesis of chronic otitis media with effusion. PMID- 11926897 TI - Inhibition of intestinal epithelial apoptosis and survival in a murine model of pneumonia-induced sepsis. AB - CONTEXT: Increased intestinal epithelial apoptosis is present in both human autopsy studies and animal models of sepsis. Whether altering gut apoptosis decreases mortality in sepsis induced by pathogenic bacteria outside the gut is unknown. OBJECTIVE: To determine if decreasing levels of intestinal cell death improves survival in a murine model of Pseudomonas aeruginosa pneumonia-induced sepsis. DESIGN AND MATERIALS: Prospective study in which transgenic mice that overexpress the antiapoptotic protein Bcl-2 in their intestinal epithelium (n = 25) and control littermates (n = 26) were subjected to intratracheal injection of P aeruginosa. MAIN OUTCOME MEASURES: Survival at 7 postoperative days, compared between the 2 groups. Secondary outcomes included quantification of gut epithelial apoptosis. RESULTS: Survival in transgenic mice that overexpress Bcl-2 in the intestinal epithelium was 40% (10/25) compared with 4% (1/26) in control littermates 7 days after intratracheal injection of P aeruginosa (P =.001), with differences in survival noted within 24 hours of surgery. Overexpression of Bcl-2 was associated with a decrease in gut epithelial apoptosis demonstrated by active caspase 3 staining, the terminal deoxynucleotidyltransferase-mediated dUTP nick end-labeling assay, and hematoxylin-eosin staining. CONCLUSIONS: In this murine model, inhibiting gut epithelial apoptosis by overexpression of Bcl-2 was associated with a survival advantage in P aeruginosa pneumonia-induced sepsis. These results suggest that intestinal epithelial apoptosis may play a role in sepsis-related mortality. PMID- 11926898 TI - Federal funding for biomedical research: commitment and benefits. PMID- 11926899 TI - The future of biomedical research: from the inventory of genes to understanding physiology and the molecular basis of disease. PMID- 11926900 TI - Basic science and translational research in JAMA. PMID- 11926907 TI - The otolaryngological manifestations of mitochondrial disease and the risk of neurodegeneration with infection. AB - OBJECTIVE: To report the nature and extent of hearing loss and other otolaryngological problems in patients with mitochondrial disease, and to document the risk of neurodegeneration with infection. DESIGN: Medical chart review and telephone interview of 40 patients with documented mitochondrial disease. SETTING: An international referral center for the diagnosis and management of mitochondrial disorders. PATIENTS: We describe 40 patients with a definitive diagnosis of mitochondrial disease. Thirty-three (82%) were younger than 15 years. RESULTS: Hearing loss was the most common clinical finding associated with mitochondrial disease. Twenty-eight (80%) of the 35 patients undergoing testing had hearing loss or significant auditory dysfunction. In 20 (57%) of these, brainstem conduction abnormalities were identified. Eight (30%) of the 27 patients had an abnormal number of recurrent upper respiratory tract infections, and 4 (50%) of these had life-threatening or neurodegenerative sequelae. Mitochondrial disease followed an episodic course, with periods of stasis or slow developmental progress, punctuated by neurodegenerative events in 18 (60%) of 30 patients. Intercurrent infection was recognized as a precipitant of neurodegenerative events in 13 (72%) of 18 patients with a history of episodic degeneration. CONCLUSIONS: Children and adults with mitochondrial disorders are at high risk for hearing loss and life-threatening complications of intercurrent infections. A constellation of audiologic abnormalities, multiorgan system involvement, and history of neuromuscular setbacks with infection strongly suggests mitochondrial disease. Knowledge of these features can lead to more rapid diagnosis and improved medical and surgical management for this special group of patients with fundamental defects in bioenergy metabolism. PMID- 11926908 TI - Is corticosteroid therapy effective for sudden-onset sensorineural hearing loss at lower frequencies? AB - OBJECTIVE: To assess the efficacy of corticosteroid therapy for sudden-onset sensorineural hearing loss at lower frequencies. DESIGN: Retrospective, nonrandomized, controlled study. PATIENTS AND METHODS: The recovery rate, recovery/improvement rate, and period between the start of therapy and improvement of symptoms were studied in 2 groups: the control group, consisting of 36 patients treated with adenosine triphosphate disodium, kallidinogenase, and cyanocobalamin between March 1, 2000, and January 31, 2001; and the corticosteroid-treated group, consisting of 42 patients treated with corticosteroids in addition to the previously mentioned drugs between April 1, 1997, and February 29, 2000. RESULTS: No significant difference (P =.83) was noted in the recovery rate between the control group (81%) and the corticosteroid treated group (79%), and the recovery/improvement rate was the same for the 2 groups (83%). In addition, there was no significant difference (P =.84) for the mean +/- SD period between the start of therapy and improvement of symptoms between the control group (3.9 +/- 2.7 days) and the corticosteroid-treated group (3.7 +/- 2.1 days). CONCLUSION: Corticosteroids were not effective for sudden onset sensorineural hearing loss at lower frequencies. PMID- 11926909 TI - Auditory and facial nerve function following surgery for cerebellopontine angle meningiomas. AB - OBJECTIVE: To investigate the postoperative auditory and facial nerve function results after cerebellopontine angle meningioma removal. DESIGN: Retrospective chart review. SETTING: Tertiary care referral center. PATIENTS: Twenty-one patients undergoing surgical removal of cerebellopontine angle meningiomas by the senior author (R.J.W.). INTERVENTIONS: Translabyrinthine or retrosigmoid approach for tumor extirpation. MAIN OUTCOME MEASURES: Postoperative auditory (pure-tone average and speech discrimination score) and facial (House-Brackmann scale) function within 1 year of follow-up. RESULTS: Twenty-three operations were performed on 21 patients. Hearing preservation through the retrosigmoid approach was attempted in 11 patients (48%). Normal hearing (class A) was preserved in 9 of 10 patients. Normal postoperative facial nerve function (House-Brackmann grade I) was conserved in 11 (65%) of 17 patients. CONCLUSIONS: This review demonstrates that successful hearing preservation is possible with meningiomas. Therefore, the retrosigmoid approach should be used whenever serviceable hearing is present preoperatively. Normal facial nerve function can also be preserved in the majority of patients. PMID- 11926910 TI - Natriuretic peptide receptors in the human endolymphatic sac. AB - OBJECTIVE: To examine human endolymphatic sac (ELS) tissue for atrial natriuretic peptide (ANP) receptor subtypes A, B, and C. DESIGN: Pilot study. METHODS: Immunohistochemical analysis of human ELS tissue specimens. The ANP receptors were characterized using the peroxidase/antiperoxidase method and polyclonal antibodies directed against each receptor subtype. The identity of the stain regarding receptor subclass was masked from the observer. Human kidney tissue known to contain all 3 receptor subtypes was used as a control. Presence of the receptor subclasses was confirmed using reverse transcriptase-polymerase chain reaction (RT-PCR) techniques. SUBJECTS: Samples of ELS tissue were obtained at autopsy from 3 fresh cadaver specimens (6 ears) and as surgical specimens from 3 patients (1 for immunohistochemical analysis and 2 for RT-PCR) undergoing acoustic neuroma resection using the translabyrinthine approach. RESULTS: The ANP type B receptors demonstrated moderate to strong reactivity in all 7 specimens, and mild to moderate staining to the ANP type C receptor was also noted. No appreciable reactivity to the ANP type A receptor was detected using immunohistochemical techniques. All 3 receptor subclasses were detected using RT PCR. CONCLUSIONS: The ANP receptors are found within the human ELS, with a predominance of ANP type B based on the intensity of staining. The ANPs may be involved in fluid homeostasis in the inner ear. Based on these findings, C-type natriuretic peptide may be a more effective peptide within the human ELS for fluid regulation because its binding affinity is virtually exclusive for the ANP type B receptor. PMID- 11926911 TI - Concomitant chemoradiotherapy in pyriform sinus carcinoma. AB - OBJECTIVES: To test the effectiveness of concurrent chemoradiotherapy in patients with pyriform sinus carcinoma and to demonstrate the feasibility of an organ preservation approach. DESIGN: Clinical trial phase 2. SETTING: University Hospital Center, St-Etienne, France. PATIENTS: The study population comprised 46 male patients with resectable stage III and IV pyriform sinus carcinoma. METHODS: Two successive chemoradiation regimens were investigated. In protocol 1 (24 patients), carboplatin was given on days 1 through 5 and 28 through 33, with an area under the curve dose of 5 mg/mL for 1 minute per day and bifractionated radiotherapy (160 rad [1.6 Gy]/fraction) delivered on days 1 through 16 and 28 through 38. A treatment break was planned on days 16 through 27. In protocol 2 (22 patients), chemotherapy was given with the same dose of carboplatin on days 1 and 21, and fluorouracil (750 mg/m(2) per day) on days 1 through 7 and 21 through 28. Radiotherapy with a single fraction of 180 rad (1.8 Gy)/d was delivered during the first 2 weeks and then 150 rad (1.5 Gy) twice a day during the next 3 weeks. MAIN OUTCOME MEASURES: Patients were evaluated for tumor response, toxic reactions, and organ preservation and survival rates. Statistical analysis of disease-free survival and overall survival was performed using the Kaplan-Meier method. RESULTS: A complete response was noted in 21 (88%) of the 24 patients following protocol 1 and 16 (73%) of the 22 patients following protocol 2. After 2 years of follow up, 16 patients (67%) (protocol 1) and 12 patients (55%) (protocol 2) retained their larynx without evidence of disease. During therapy, 15 patients (63%) (protocol 1) and 19 patients (86%) (protocol 2) required unplanned hospitalization for toxic effects. The overall survival and disease free survival rates at 2 years were 58% (protocol 1) vs 53% (protocol 2) and 39% (protocol 1) vs 41% (protocol 2) (P =.80), respectively. CONCLUSION: Concomitant chemotherapy and bifractionated radiotherapy, although toxic, leads to good locoregional control and therefore to a significant level of laryngeal preservation. PMID- 11926912 TI - Assessment of the morbidity and complications of total thyroidectomy. AB - OBJECTIVE: To determine the incidence and predictive factors for complications after total thyroidectomy. DESIGN: Cross-sectional analysis of a national database on total thyroidectomy cases. METHODS: The National Hospital Data Survey database was examined and all cases of total thyroidectomy performed during 1995 to 1999 were extracted. In addition to demographic information, postoperative complications including hypocalcemia, recurrent laryngeal nerve paralysis, wound complications, and medical morbidities were identified. Statistical analysis was conducted to determine potential predictive factors for postoperative complications. RESULTS: A total of 517 patients were identified (mean age, 48.3 years). The most common indications for total thyroidectomy were thyroid malignancy and goiter (73.9% of cases). Eighty-one patients (15.7%) underwent an associated nodal dissection along with total thyroidectomy, and 16 patients (3.1%) underwent parathyroid reimplantation. The mean length of stay was 2.5 days (95% confidence interval, 2.3-2.8 days). The incidence of postoperative wound hematoma was 1.0%, wound infection was 0.2%, and mortality rate was 0.2%. The incidence of postoperative hypocalcemia was 6.2%. Younger age was statistically associated with an increased incidence of hypocalcemia (P =.002, t test), whereas sex (P =.48), indication for surgery (P =.32), parathyroid reimplantation (P>.99), and associated neck dissection (P =.21) were not. The mean length of stay was 2.5 days and was unaffected by occurrence of postoperative hypocalcemia. The incidences of unilateral and bilateral vocal cord paralyses were 0.77% and 0.39%, respectively. CONCLUSIONS: Postoperative hypocalcemia is the most common immediate surgical complication of total thyroidectomy. Other complications, including recurrent laryngeal nerve paralysis, can be expected at rates approximating 1%. PMID- 11926913 TI - Prognostic value of CD44 variant 6 in laryngeal epidermoid carcinomas. AB - BACKGROUND: CD44 variant exon 6 (v6) belongs to a family of transmembrane glycoproteins involved in cell adhesion. OBJECTIVES: To determine the prognostic role of CD44v6 in laryngeal cancer and to examine its relation with other clinicopathologic prognostic factors. DESIGN: A retrospective cohort study was designed with 93 laryngeal cancer cases. They were selected randomly from patients treated with laryngectomy between January 1, 1983, and December 31, 1993. SETTING: Faculty of Medicine, Hacettepe University, Ankara, Turkey. PATIENTS: The ages of the patients ranged from 31 to 73 years. Eighty-eight patients were men and 5 were women. Three had stage I, 33 had stage II, 27 had stage III, and 30 had stage IV disease at the time of surgery. INTERVENTION: Histological sections of tumors and metastatic lymph nodes were reevaluated for several histopathological factors. Sections were stained using anti-CD44v6 monoclonal antibody by immunohistochemical methods. RESULTS: CD44v6 expression was seen only in the lower one third of the normal squamous epithelium but in all layers of dysplasia and in situ carcinoma. Besides a general evaluation of tumor staining, immunostaining was evaluated separately for cell groups located in the center of neoplastic islands (nonbasal cells), at the periphery of the neoplastic islands (basal cells), and at the infiltration zones (marginal cells). Decreased disease-free survival was noted when there was extensive staining in the general evaluation and in cases with extensive staining in marginal and nonbasal cells (P =.03). Using Cox regression analysis, the greatest dimension of the largest metastatic lymph node and extensive expression of CD44v6 in nonbasal tumor cells were independent prognostic factors. CONCLUSION: Our results suggest that CD44v6 expression is an important prognostic factor in laryngeal cancer. PMID- 11926914 TI - Topical mitomycin as an adjunct to choanal atresia repair. AB - OBJECTIVE: To evaluate the use of topical mitomycin in choanal atresia repair to reduce the development of granulation tissue and cicatrix. DESIGN AND SETTING: Retrospective case series in 2 tertiary care centers. PATIENTS: Twenty patients with either unilateral or bilateral congenital choanal atresia underwent repair using the transnasal endoscopic approach, the transpalatal approach, or both. INTERVENTIONS: The surgeons favor the use of the endoscopic transnasal drillout technique for all unilateral cases of choanal atresia and for selected bilateral cases. We describe our experience and treatment paradigm for these 20 patients (15 with unilateral atresia, 5 with bilateral atresia). Topical application of mitomycin was used, and in some cases postoperative stenting, for a period of 1 to 2 weeks. In 8 cases, a second application of mitomycin was used. Follow-up ranged from 3 months to 2 years (mean, 9 months). OUTCOME MEASURE: The patency of the choanae without respiratory distress or nasal drainage, as assessed by endoscopic evaluation, determined a successful repair. RESULTS: Of the 20 patients, 17 retained patent airways. Three patients experienced improvement from a total atresia to a narrowed, stenotic choana. CONCLUSIONS: The use of mitomycin as an adjunct to the surgical repair of choanal atresia may offer improved patency with a decreased need for stenting, dilatations, and revision surgery. Newer endoscopic techniques with powered instrumentation further enhance the safety and efficacy in the repair of choanal atresia. PMID- 11926915 TI - The role of mitomycin in the prevention and treatment of scar formation in the pediatric aerodigestive tract: friend or foe? AB - OBJECTIVE: To evaluate the role of mitomycin in the prevention and treatment of scar formation in the pediatric aerodigestive tract. DESIGN: Prospective study; institutional review board-approved clinical trial. SETTING: Tertiary care pediatric medical center. PATIENTS: Fifteen patients; choanal atresia in 5 patients, airway stenosis in 8 patients, hypopharyngeal stenosis in 1 patient, and esophageal stenosis in 1 patient. OUTCOME: The efficacy and safety of mitomycin in the prevention of scar formation. INTERVENTION: All patients underwent surgical repair of the stenotic area, followed by topical application of mitomycin (1 mL of 0.4 mg/mL) for 4 minutes. RESULTS: Ten patients (67%) showed major improvement, 4 patients (27%) showed minor improvement, and 1 patient (7%) showed no improvement. CONCLUSION: Topical application of mitomycin can play an effective role in the prevention and treatment of scar formation in the aerodigestive tract. PMID- 11926917 TI - Clinical assessment of retronasal olfactory function. AB - OBJECTIVES: To develop a test kit for the simple assessment of retronasal olfactory function and to compare orthonasal and retronasal olfactory function in healthy subjects and patients with olfactory disorders. DESIGN AND PATIENTS: We tested 230 individuals with normosmia, hyposmia, and anosmia using grocery available powders. Initially, 30 different substances were investigated. Subjects identified each substance using a list with 4 verbal items (forced choice). After preliminary experiments, 20 items were selected according to the degree to which they were identified by normosmic and anosmic subjects. Orthonasal olfactory function was assessed psychophysically using "sniffin' sticks," which includes tests for odor identification, discrimination, and butanol odor thresholds. In addition, anosmia was confirmed electrophysiologically by means of olfactory evoked potentials. RESULTS: In healthy subjects, there was a test-retest reliability correlation of r(27) = 0.76 for retronasal olfactory function, which is similar to other odor identification tests. Retronasal testing in normosmic subjects allowed for the discrimination of sex-related differences, with women scoring higher than men (P =.007), and the identification of a slight decrease with age (r(120) = -0.20; P =.03). Orthonasal and retronasal identification of odors was found to correlate (r(86) = 0.78; P<.001). Retronasal testing allowed for the discrimination between normosmia, hyposmia, and anosmia (P<.001). In addition, retronasal performance of anosmic patients appeared to improve with duration of anosmia (P =.03). No difference was found between patients with anosmia of different origin. CONCLUSION: Results of the present investigation indicate that the assessment of retronasal olfactory function is possible using oral stimulus presentation. PMID- 11926916 TI - Orbitofacial masses in children: an endoscopic approach. AB - OBJECTIVE: To describe an endoscopic approach for pediatric orbitofacial masses. DESIGN: A retrospective medical chart review. SETTING: Tertiary-care children's hospital. PARTICIPANTS: Patients (4 boys, 7 girls) ranged in age from 6 months to 11 years. All children underwent endoscopic excision of an orbitofacial mass. INTERVENTION: A single port approach was used in all but the initial case. The scalp incision was placed approximately 2.0 cm behind the frontal hairline. A subgaleal dissection was performed to minimize risk of nerve injury. Under endoscopic visualization, the mass was resected. MAIN OUTCOME MEASURES: Ability to successfully excise the mass endoscopically, and the incidence of complication. RESULTS: All lesions were successfully resected endoscopically. The surgical time varied from 30 to 105 minutes (mean, 50.5 minutes). Pathologic examination revealed 10 dermoid cysts and 1 neurofibroma. Two children had transient frontalis branch palsies that resolved spontaneously. There was 1 unilateral frontal hypoesthesia in the patient with the neurofibroma (an expected result). There were no other complications. CONCLUSIONS: An endoscopic approach to pediatric orbitofacial tumors is safe and effective. Although the risk of nerve injury may be higher, a thorough knowledge of frontotemporal anatomy and careful dissection will minimize this risk. The distinct advantage of an endoscopic approach is the absence of any facial scar in these young patients. PMID- 11926918 TI - Mucociliary transport and histologic characteristics of the mucosa of deviated nasal septum. AB - OBJECTIVE: To investigate differences in mucociliary clearance, histologic characteristics, and surface structure of the bilateral nasal septal mucosa in patients with nasal septal deviation. DESIGN: Mucociliary transport was measured by saccharin clearance time in both nasal cavities of 20 patients with nasal septal deviation. Their septal mucosae were taken during septoplasty, and the ciliary population was studied by scanning electron microscopy. Histologic differences in the lamina propria of septal mucosae were compared under a light microscope. RESULTS: The concave side showed longer saccharin clearance time than the convex side and revealed much more severe loss of cilia. Inflammatory cells more heavily infiltrated the concave side, and seromucinous glands were less densely distributed. CONCLUSION: Concave-side septal mucosae have impaired mucociliary transport, presumably due to ciliary loss, increased inflammation, and decreased density of the glandular acini. PMID- 11926919 TI - Powered instrumentation in the treatment of recurrent respiratory papillomatosis: an alternative to the carbon dioxide laser. AB - OBJECTIVE: To assess the advantages of powered instrumentation vs the carbon dioxide laser in treating patients with juvenile-onset recurrent respiratory papillomatosis. DESIGN: A retrospective study. SETTING: Tertiary care children's hospital. PATIENTS: Patients operated on for juvenile-onset recurrent respiratory papillomatosis between January 1, 1999, and December 31, 2000. Papillomas were excised using the microdebrider in one group and the carbon dioxide laser in the second group. INTERVENTIONS: Direct laryngoscopy and bronchoscopy, suspension microlaryngoscopy, and excision of papillomas by the carbon dioxide laser or the microdebrider. MAIN OUTCOME MEASURES: Operative time and postoperative complications. RESULTS: Seventy-three operations were performed (23 with the laser and 50 with the microdebrider). Sixteen patients were included, 10 with active disease and 5 with disease in remission; 1 was lost to follow-up. They had a mean age of 3.75 years, and the male-female ratio was 7:9. The patients presented mostly with hoarseness (13 [81%]). Four (25%) had soft tissue complications with the laser. The microdebrider was less time-consuming than the laser, although those treated with the microdebrider had more active disease. No factor could be used to measure treatment outcome due to disease variability. Those who were older, female, and African American tended to have less severe manifestations of disease. CONCLUSIONS: The microdebrider proved to be less time consuming than the carbon dioxide laser when used in patients with juvenile-onset recurrent respiratory papillomatosis. Soft tissue complications were nonexistent. In addition to safety, the microdebrider is more appealing to the surgeon, anesthesiologist, and parents, especially because these children often need subsequent surgical procedures. PMID- 11926920 TI - Laser-assisted uvulopalatoplasty for the management of obstructive sleep apnea: myths and facts. AB - OBJECTIVE: To assess medium- to long-term subjective and objective results of laser-assisted uvulopalatoplasty (LAUP) for patients with obstructive sleep apnea. DESIGN: A nonrandomized prospective before-after trial. SUBJECTS AND INTERVENTIONS: Twenty-six patients underwent LAUP by means of vertical trenches along either side of the uvula and reduction of the uvula. MAIN OUTCOME MEASURES: Subjective analysis included a preoperative and 2 postoperative evaluations of the state of snoring: 4 weeks and a mean +/- SD of 12.3 +/- 9.1 months after completion of treatment. In addition, a score on 5 other sleep-related symptoms was recorded before treatment and after 12.3 +/- 9.1 months; at that time, patients also estimated their overall satisfaction with the procedure. Objective analysis included preoperative polysomnographic studies that were repeated postoperatively. RESULTS: A significant decline in snoring improvement from 88% (23/26) to 65% (17/26) was recorded; furthermore, the state of snoring worsened from 4% (1/26) to 12% (3/26). Reevaluation of 5 other sleep-related symptoms after completion of LAUP uncovered a 50% improvement rate (13/26), and a 15% (4/26) worsening rate. Overall satisfaction from the procedure was 58% (15/26). Postoperative objective studies revealed that only 31% (8/26) of the procedures were successful, while 31% were associated with worsening of respiratory disturbance index. Fifty-four percent (14/26) of the patients had a sensation of pharyngeal dryness. In addition, 1 patient developed velopharyngeal stenosis. CONCLUSIONS: The favorable subjective short-term results of LAUP deteriorated in time. Postoperative polysomnography revealed that LAUP might lead to deterioration of existing apnea. These findings are probably related to velopharyngeal narrowing and progressive palatal fibrosis inflicted by the laser beam. PMID- 11926921 TI - Usefulness of uvulopalatopharyngoplasty with genioglossus and hyoid advancement in the treatment of obstructive sleep apnea. AB - OBJECTIVE: To evaluate the usefulness of uvulopalatopharyngoplasty plus mandibular osteotomy with genioglossus and hyoid advancement in the treatment of obstructive sleep apnea syndrome (OSAS). DESIGN: Prospective study of 20 consecutive patients with OSAS. SETTING: University medical center. PATIENTS AND INTERVENTIONS: Twenty OSAS patients with multilevel upper airway obstruction who refused continuous positive airway pressure treatment. All patients were evaluated before and 6 months after surgery by clinical history, the Epworth Sleepiness Scale, physical examination, fiberoptic nasopharyngoscopy combined with the Muller maneuver, cephalometric analysis, nocturnal polysomnography, and a second-night polysomnography with upper airway pressure recording during sleep. Surgery procedures were uvulopalatopharyngoplasty plus mandibular osteotomy with genioglossus and hyoid advancement. Surgical successful outcome was defined as an apnea-hypopnea index (AHI) lower than 20 plus subjective resolution of daytime symptoms. MAIN OUTCOME MEASURE: Surgical success rate. RESULTS: Mean +/- SD AHI decreased from 60.5 +/- 16.5 to 44.6 +/- 27(P =.007), and CT90 (percentage of time with oxyhemoglobin saturation below 90%) decreased from 39.5% +/- 26% to 25.1% +/- 26.4% (P =.002). The overall surgical success rate was 35% but increased to 57% in patients with moderate OSAS (AHI, 41-60) and to 100% in mild OSAS (AHI, 21-40). In the group of severe OSAS, the success rate was 9%. Predictors of surgical outcome success were the AHI, CT90, stages 2 and 3-4 sleep percentages, and the cephalometric ANB angle (angle formed from the deepest point on the maxillary outer contour to the nasion to the deepest point on the outer mandibular contour). CONCLUSION: Patients with mild and moderate OSAS and multilevel obstruction in the upper airway may benefit from uvulopalatopharyngoplasty plus genioglossus and hyoid advancement. PMID- 11926922 TI - Primary description of a new entity, renal cell-like carcinoma of the nasal cavity: van Meegeren in the house of Vermeer. AB - BACKGROUND: Few sinonasal malignancies can manifest, histologically, as clear cell neoplasia. The most likely such tumor to be encountered is metastatic renal cell carcinoma. Primary sinonasal tumors that can appear as clear cell malignancies include squamous cell carcinoma and mucoepidermoid carcinoma. Primary salivary clear cell carcinoma occurs almost exclusively in the oral cavity and has not been described in the nasal cavity. OBJECTIVE: To report a unique sinonasal clear cell malignancy that mimicked metastatic renal carcinoma. STUDY DESIGN: Case report. OUTCOME MEASUREMENTS: Radiography, histology, histochemistry, immunohistochemistry, and electron microscopy. RESULTS: Histologically, the tumor was identical to renal cell carcinoma. No evidence of renal malignancy was found by abdominal computed tomographic scan or gadolinium enhanced magnetic resonance imaging. Histochemistry confirmed the presence of tumor glycogen but no mucin. Immunohistochemistry confirmed strong expression of low- and high-molecular-weight keratin and S100, and no vimentin expression. Electron microscopy showed tumor myofibroblastic differentiation and cytoplasmic glycogen, neutral lipid vacuoles, and cholesterol. CONCLUSIONS: There was no clinical evidence of renal cell carcinoma. The immunohistochemical and ultrastructural findings were inconsistent with the diagnosis of renal cell carcinoma and showed features also inconsistent with the diagnosis of primary salivary clear cell carcinoma. We therefore conclude that this tumor represents a new and distinct entity, notable in its presentation as a "counterfeit renal cell carcinoma." PMID- 11926923 TI - Lipomatous variant of nasopharyngeal angiofibroma: a case report. AB - Nasopharyngeal angiofibroma is an uncommon benign vascular tumor that occurs in adolescent boys and young men. We describe an unusual histological variant of this entity characterized by a prominent adipose tissue component. The patient was a 35-year-old man with a 20-year history of nasal obstruction. Results of physical examination and angiography demonstrated a nasal mass consistent with a nasopharyngeal angiofibroma. We discuss the differential diagnoses pathologically for this lesion. PMID- 11926924 TI - Pathology quiz case 1: nasoalveolar cyst. PMID- 11926925 TI - Pathology quiz case 2: secondary hyperparathyroidism and brown tumor of the mandible. PMID- 11926926 TI - Radiology quiz case 1: persistent stapedial artery, left ear. PMID- 11926927 TI - Radiology quiz case 2: metastatic calcifications of the middle and external ear and osteitis fibrosa of the temporal bones as a result of secondary hyperparathyroidism. PMID- 11926928 TI - Papillary carcinoma in a thyroglossal duct remnant. AB - For papillary carcinoma in a thyroglossal duct remnant, complete wide excision of the remnant is adequate treatment. PMID- 11926931 TI - Neuronal substrate of the saccadic inhibition deficit in schizophrenia investigated with 3-dimensional event-related functional magnetic resonance imaging. AB - BACKGROUND: Several studies have shown that the ability to suppress automatic saccadic eye movements is impaired in patients with schizophrenia as well as in their first-degree relatives, and suggest that this impairment is a potential vulnerability marker for schizophrenia. The neurobiological mechanisms underlying normal saccade production and inhibition, revealed in primate studies, indicate that the impairment may result from a failure of the oculomotor system to effectively exert inhibitory control over brainstem structures. Functional localization of the affected brain structure(s) potentially provides a physiological measure for the investigation of vulnerability markers in schizophrenia. METHODS: The hemodynamic response to discrete visual stimuli was measured during prosaccades (saccades toward a peripheral stimulus), antisaccades (saccades toward a position opposite to a peripheral stimulus), and active fixation (holding fixation and ignoring a peripheral stimulus) in 16 patients with schizophrenia receiving atypical neuroleptics and 17 healthy control subjects using an event-related functional magnetic resonance imaging task design. RESULTS: Brain responses were detected in the frontal and parietal regions of the oculomotor system in all 3 tasks. Patients made more errors during inhibition tasks and exhibited a selective failure to activate the striatum during the inhibition of saccades. In other regions that were active during inhibition, specifically the supplementary and frontal eye fields, no difference was found between patients and control subjects. CONCLUSIONS: A frontostriatal network is engaged in the suppression of automatic eye movements. The results indicate that abnormalities in this network, rather than the selective dysfunction of prefrontal brain regions, underlie the saccade inhibition deficit in schizophrenia. PMID- 11926932 TI - Association between smaller left posterior superior temporal gyrus volume on magnetic resonance imaging and smaller left temporal P300 amplitude in first episode schizophrenia. AB - BACKGROUND: In chronic schizophrenia, the P300 is broadly reduced and shows a localized left temporal deficit specifically associated with reduced gray matter volume of the left posterior superior temporal gyrus (STG). In first-episode patients, a similar left temporal P300 deficit is present in schizophrenia, but not in affective psychosis. The present study investigated whether the left temporal P300-left posterior STG volume association is selectively present in first-episode schizophrenia. METHOD: P300 was recorded as first-episode subjects with schizophrenia (n = 15) or affective psychosis (n = 18) or control subjects (n = 18) silently counted infrequent target tones amid standard tones. High resolution spoiled gradient-recalled acquisition magnetic resonance images provided quantitative measures of temporal lobe gray matter regions of interest. RESULTS: Patients with first-episode schizophrenia displayed a reversed P300 temporal area asymmetry (smaller on the left), while magnetic resonance imaging showed smaller gray matter volumes of left posterior STG relative to control subjects and patients with affective psychosis (15.4% and 11.0%, respectively), smaller gray matter volumes of left planum temporale (21.0% relative to both), and a smaller total Heschl's gyrus volume (14.6% and 21.1%, respectively). Left posterior STG and the left planum temporale, but not other regions of interest, were specifically and positively correlated (r>0.5) with left temporal P300 voltage in patients with schizophrenia but not in patients with affective psychosis or in control subjects. CONCLUSION: These results suggest that the left temporal P300 abnormality specifically associated with left posterior STG gray matter volume reduction is present at the first hospitalization for schizophrenia but is not present at the first hospitalization for affective psychosis. PMID- 11926933 TI - Structural brain differences between never-treated patients with schizophrenia, with and without dyskinesia, and normal control subjects: a magnetic resonance imaging study. AB - BACKGROUND: In south India, abnormal movements indistinguishable from tardive dyskinesia have been observed in chronically ill patients with schizophrenia who have never received antipsychotic medication. The present study, using magnetic resonance imaging, examines brain structure in such patients, in those without dyskinesia, and in normal control subjects. METHODS: Chronically ill patients with schizophrenia with and without dyskinesia and controls were identified in villages south of Chennai, India (each group, n = 31). Patients' mental state was assessed by the Positive and Negative Syndrome Scale for schizophrenia, dyskinesia by the Abnormal Involuntary Movements Scale, and parkinsonism by the Simpson and Angus scale. In patients and controls, magnetic resonance imaging measured the volume of the caudate and lentiform nuclei and the lateral ventricle hemisphere ratio. RESULTS: The left lentiform nucleus was significantly (11%) larger in patients with dyskinesia compared with controls, and the right lateral ventricle-hemisphere ratio was significantly (33%) larger in patients without dyskinesia compared with controls. In all 3 groups, there were significant positive correlations between age and ventricle-hemisphere ratio. In controls, but not in patients, there were significant negative correlations between age and the volume of the caudate and lentiform nuclei. CONCLUSIONS: Never-treated patients with dyskinesia may have striatal pathologic conditions and may represent a subgroup of patients with schizophrenia; in those without abnormal movements, cortical atrophy is more apparent. The schizophrenic process may interfere with normal age-related anatomical changes in the basal ganglia. PMID- 11926934 TI - Abnormalities in glucose regulation during antipsychotic treatment of schizophrenia. AB - BACKGROUND: Hyperglycemia and type 2 diabetes mellitus are more common in schizophrenia than in the general population. Glucoregulatory abnormalities have also been associated with the use of antipsychotic medications themselves. While antipsychotics may increase adiposity, which can decrease insulin sensitivity, disease- and medication-related differences in glucose regulation might also occur independent of differences in adiposity. METHODS: Modified oral glucose tolerance tests were performed in schizophrenic patients (n = 48) receiving clozapine, olanzapine, risperidone, or typical antipsychotics, and untreated healthy control subjects (n = 31), excluding subjects with diabetes and matching groups for adiposity and age. Plasma was sampled at 0 (fasting), 15, 45, and 75 minutes after glucose load. RESULTS: Significant time x treatment group interactions were detected for plasma glucose (F(12,222) = 4.89, P<.001) and insulin (F(12,171) = 2.10, P =.02) levels, with significant effects of treatment group on plasma glucose level at all time points. Olanzapine-treated patients had significant (1.0-1.5 SDs) glucose elevations at all time points, in comparison with patients receiving typical antipsychotics as well as untreated healthy control subjects. Clozapine-treated patients had significant (1.0-1.5 SDs) glucose elevations at fasting and 75 minutes after load, again in comparison with patients receiving typical antipsychotics and untreated control subjects. Risperidone-treated patients had elevations in fasting and postload glucose levels, but only in comparison with untreated healthy control subjects. No differences in mean plasma glucose level were detected when comparing risperidone treated vs typical antipsychotic-treated patients and when comparing typical antipsychotic-treated patients vs untreated control subjects. CONCLUSION: Antipsychotic treatment of nondiabetic patients with schizophrenia can be associated with adverse effects on glucose regulation, which can vary in severity independent of adiposity and potentially increase long-term cardiovascular risk. PMID- 11926935 TI - Evidence for impaired cortical inhibition in schizophrenia using transcranial magnetic stimulation. AB - BACKGROUND: Cortical inhibition (CI) deficits have been proposed as a pathophysiologic mechanism in schizophrenia. This study employed 3 transcranial magnetic stimulation (TMS) paradigms to assess CI in patients with schizophrenia. Paired-pulse TMS involves stimulating with a lower-intensity pulse a few milliseconds before a higher-intensity pulse, thereby inhibiting the size of the motor evoked potential produced by the higher-intensity pulse. In the cortical silent period paradigm, inhibition is reflected by the silent period duration (ie, the duration of electromyographic activity cessation following a TMS-induced motor evoked potential). Transcallosal inhibition involves stimulation of the contralateral motor cortex several milliseconds prior to stimulation of the ipsilateral motor cortex, inhibiting the size of the motor evoked potential produced by ipsilateral stimulation. METHODS: We measured CI using these 3 paradigms in 15 unmedicated patients with schizophrenia (14 medication-naive and 1 medication-free for longer than 1 year) (13 were in the transcallosal inhibition paradigm), 15 medicated patients with schizophrenia (11 taking olanzapine, 1 risperidone, 1 quetiapine, 1 methotrimeprazine + perphenazine, 1 quetiapine + loxapine), and 15 healthy controls. RESULTS: Unmedicated patients demonstrated significant CI deficits compared with healthy controls across all inhibitory paradigms whereas medicated patients did not (at all inhibitory intervals, paired-pulse TMS: controls = 59.9%, medicated = 44.3%, unmedicated = 28.7%; cortical silent period: controls = 55.0 milliseconds, medicated = 60.4 milliseconds, unmedicated = 39.7 milliseconds; transcallosal inhibition: controls = 33.6%, medicated = 23.7%, unmedicated = 10.4%; P<.05). CONCLUSIONS: These results suggest that schizophrenia is associated with deficits in CI and that antipsychotic medications may increase CI. PMID- 11926936 TI - Cost and cost-effectiveness of hospital vs residential crisis care for patients who have serious mental illness. AB - BACKGROUND: This study evaluates the cost and cost-effectiveness of a residential crisis program compared with treatment received in a general hospital psychiatric unit for patients who have serious mental illness in need of hospital-level care and who are willing to accept voluntary treatment. METHODS: Patients in the Montgomery County, Maryland, public mental health system (N = 119) willing to accept voluntary acute care were randomized to the psychiatric ward of a general hospital or a residential crisis program. Unit costs and service utilization data were used to estimate episode and 6-month treatment costs from the perspective of government payors. Episodic symptom reduction and days residing in the community over the 6 months after the episode were chosen to represent effectiveness. RESULTS: Mean (SD) acute treatment episode costs was $3046 ($2124) in the residential crisis program, 44% lower than the $5549 ($3668) episode cost for the general hospital. Total 6-month treatment costs for patients assigned to the 2 programs were $19,941 ($19,282) and $25,737 ($21,835), respectively. Treatment groups did not differ significantly in symptom improvement or community days achieved. Incremental cost-effectiveness ratios indicate that in most cases, the residential crisis program provides near-equivalent effectiveness for significantly less cost. CONCLUSIONS: Residential crisis programs may be a cost effective approach to providing acute care to patients who have serious mental illness and who are willing to accept voluntary treatment. Where resources are scarce, access to needed acute care might be extended using a mix of hospital, community-based residential crisis, and community support services. PMID- 11926937 TI - Parental major depression and the risk of depression and other mental disorders in offspring: a prospective-longitudinal community study. AB - BACKGROUND: This article examines associations between DSM-IV depressive disorders, their natural course, other psychopathology, and parental major depression in a community sample of adolescents and young adults. METHODS: Baseline and 4-year follow-up data were used from the Early Developmental Stages of Psychopathology Study, a prospective-longitudinal community study of adolescents and young adults. Results are based on 2427 subjects who completed the follow-up and for whom diagnostic information for both parents was available. DSM-IV mental disorders in respondents were assessed using the Munich-Composite International Diagnostic Interview. Information on depression in parents was collected as family history information from the respondents and from diagnostic interviews with parents of the younger cohort. RESULTS: Offspring with 1 (odds ratio [OR], 2.7; 95% confidence interval [CI], 2.1-3.5) or 2 affected parents (OR, 3.0; 95% CI, 2.2-4.1) had an increased risk for depression. They also had a higher risk for substance use (1 parent affected: OR, 1.4; 95% CI, 1.1-1.7; both parents affected: OR, 1.4; 95% CI, 1.0-1.8) and anxiety disorders (1 parent affected: OR, 1.6; 95% CI, 1.3-1.9; both parents affected: OR, 2.1; 95% CI, 1.6 2.8). There were no differences whether mother or father was affected. Parental depression was associated with an earlier onset and a more malignant course (severity, impairment, recurrence) of depressive disorders in offspring. CONCLUSIONS: Major depression in parents increases the overall risk in offspring for onset of depressive and other mental disorders and influences patterns of the natural course of depression in the early stages of manifestation. PMID- 11926938 TI - Effects of major depression on remission and relapse of substance dependence. AB - BACKGROUND: The effects of major depressive disorder (MDD) on the course of substance dependence may differ depending on the temporal relationship of depression to dependence. We investigated the effects of MDD on the outcome of substance dependence under 3 circumstances: (1) lifetime onset of MDD prior to lifetime onset of dependence onset, (2) current MDD occurring during a period of abstinence, and (3) current MDD during substance use that exceeded the expected effects of intoxication or withdrawal. METHODS: A sample of 250 inpatients with DSM-IV cocaine, heroin, and/or alcohol dependence were followed up at 6, 12, and 18 months. The Psychiatric Research Interview for Substance and Mental Disorders (PRISM) was used to make DSM-IV diagnoses. Using Cox proportional hazards models, stable remissions (those lasting at least 26 weeks) from DSM-IV cocaine, heroin, and/or alcohol dependence and from use were studied, as well as subsequent relapses of dependence and use. RESULTS: Patients with current substance-induced MDD were less likely to remit from dependence (adjusted hazards ratio, 0.11) than patients with no baseline MDD. A history of MDD prior to lifetime onset of substance dependence also reduced the likelihood of remission relative to the absence of such a history (adjusted hazard ratio, 0.49). Major depressive disorder during sustained abstinence predicted dependence relapse (adjusted hazards ratio, 3.07) and substance use after hospital discharge compared with those without abstinence MDD (adjusted hazards ratio, 1.45). CONCLUSION: The timing of depressive episodes relative to substance dependence served as an important factor in the remission and relapse of substance dependence and substance use. PMID- 11926939 TI - A randomized placebo-controlled trial of fluoxetine in body dysmorphic disorder. AB - BACKGROUND: Research on the pharmacotherapy of body dysmorphic disorder (BDD), a common and often disabling disorder, is limited. Available data suggest that this disorder may respond to serotonin reuptake inhibitors. However, no placebo controlled treatment studies of BDD have been published. METHODS: Seventy-four patients with DSM-IV BDD or its delusional variant were enrolled and 67 were randomized into a placebo-controlled parallel-group study to evaluate the efficacy and safety of fluoxetine hydrochloride. After 1 week of single-blind placebo treatment, patients were randomized to receive 12 weeks of double-blind treatment with fluoxetine or placebo. Outcome measures included the Yale-Brown Obsessive Compulsive Scale Modified for Body Dysmorphic Disorder (BDD-YBOCS) (the primary outcome measure), the Clinical Global Impressions Scale, the Brown Assessment of Beliefs Scale, and other measures. RESULTS: Results of the BDD YBOCS indicated that fluoxetine was significantly more effective than placebo for BDD beginning at week 8 and continuing at weeks 10 and 12 (F(1,64) = 16.5; P<.001). The response rate was 18 (53%) of 34 to fluoxetine and 6 (18%) of 33 to the placebo (chi(2)(1) = 8.8; P=.003). The BDD symptoms of delusional patients were as likely as those of nondelusional patients to respond to fluoxetine, and no delusional patients responded to the placebo. In the sample as a whole, treatment response was independent of the duration and severity of BDD and the presence of major depression, obsessive-compulsive disorder, or a personality disorder. Fluoxetine was generally well tolerated. CONCLUSION: Fluoxetine is safe and more effective than placebo in delusional and nondelusional patients with BDD. PMID- 11926940 TI - The surgeon's art. PMID- 11926941 TI - Late outcomes after laparoscopic surgery for gastroesophageal reflux. AB - HYPOTHESIS: Patients still have symptoms following laparoscopic antireflux surgery and require ongoing treatment. DESIGN: Mailed survey. SETTING: Academic tertiary care center. PATIENTS: Of 247 consecutive adults, 197 (80% response) who underwent laparoscopic fundoplication for gastroesophageal reflux disease in the prior 1 to 5 years. MAIN OUTCOME MEASURES: Gastrointestinal symptoms (frequency and bother), actions taken to treat these symptoms (medications and dietary and lifestyle changes), and assessment of surgery. RESULTS: The mean age of the respondents was 51.1 years, and 52% were men. The median time since surgery was 2.6 years. Overall, 28% reported typical reflux symptoms (heartburn or regurgitation), but only 5% were bothered "a lot" or "terribly" by them. While 65% reported other gastrointestinal symptoms (bloating or dysphagia) that may be related to gastroesophageal reflux disease or to surgery, only 19% were bothered a lot or terribly by them. About half of the respondents reported taking at least 1 of the following actions for their symptoms: 6% take frequent over-the-counter medications, 13% take daily prescription acid-reducing medications, 41% make lifestyle changes (eg, eating smaller meals), and 44% avoid certain foods (eg, carbonated beverages). Nevertheless, 90% believed their surgery was working well. CONCLUSIONS: In 1 to 5 years after laparoscopic antireflux surgery, many patients report gastrointestinal symptoms and take action to control these symptoms. Most, however, believe their surgical treatment is working well. PMID- 11926942 TI - Is the use of a bougie necessary for laparoscopic Nissen fundoplication? AB - HYPOTHESIS: Esophageal intubation with a bougie during laparoscopic Nissen fundoplication (LNF) is commonly used to prevent an excessively tight wrap. However, a bougie may cause intraoperative gastric and esophageal perforations. We hypothesized that LNF is safe and effective when performed without a bougie. DESIGN: Retrospective review of 102 consecutive patients who underwent LNF without a bougie. SETTING: Tertiary care university hospital. PATIENTS: All patients presented with symptoms of reflux disease. Mean (+/- SD) percentage of time with pH of less than 4 was 12.6% +/- 9.4%. Mean DeMeester score was 47.8. Mean (+/- SD) resting lower esophageal sphincter pressure was 15.0 +/- 9.4 mm Hg. Mean (+/- SD) distal esophageal amplitude was 69.4 +/- 39.2 mm Hg. INTERVENTION: During LNF, we obtained 2 to 3 cm of intra-abdominal esophagus, divided all short gastric vessels, reapproximated the crura, and performed a loose 360 degrees fundoplication without a bougie. MAIN OUTCOME MEASURES: Postoperative rates of dysphagia, gas bloat, and recurrent reflux. RESULTS: In the early postoperative period, 50 patients (49.0%) complained of mild, 11 (10.8%) of moderate, and 7 (6.9%) of severe dysphagia. Average (+/- SD) duration of early dysphagia was 4.6 +/- 2.1 weeks. Dysphagia resolved in 61 (89.7%) of 68 patients within 6 weeks. Late resolution of dysphagia was noted in 4 (5.8%) patients. Three patients were successfully treated with esophageal dilatations. Persistent dysphagia was found in 1 patient. Thirty patients (29.4%) had transient gas bloat. Mild persistent reflux, requiring daily medication, was noted in 5 (4.9%) patients. CONCLUSIONS: Performance of LNF without a bougie offers a safe and effective therapy for gastroesophageal reflux disease. While avoiding the potential risks for gastric and esophageal injury, it may provide low rates of long-term postoperative dysphagia and reflux recurrence. PMID- 11926944 TI - The Vermont colorectal cancer project: self-portrait. AB - HYPOTHESIS: A prospective statewide quality assessment and improvement project requiring active participation and case entry by surgeons is feasible provided that confidentiality and peer review protection are provided. DESIGN: Inception cohort. SETTING: Acute-care hospitals in Vermont. PATIENTS: Consecutive series of 364 patients undergoing elective surgery for colorectal cancer between April 1, 1999, and March 31, 2001, who were prospectively entered into a database created by the Vermont Chapter of the American College of Surgeons under peer review protection from the Vermont Program for Quality Health Care. INTERVENTION: Surgery for invasive colorectal cancer. MAIN OUTCOME MEASURES: Case entry compliance, surgical complications, length of stay, demographics, cancer-specific characteristics, and use of adjuvant therapy. RESULTS: The calculated case entry compliance rate was 78%. There were 7 deaths (2%) and 45 major complications in 39 patients (12.3%). All patients were offered referral for adjuvant therapy when appropriate based on National Institutes of Health Consensus Conference standards. Mean age was 68.7 years, and 52% of cancers occurred in women. The most common site of cancer was the right colon (36.6% of patients), and only 47.9% of malignancies were in the rectum or sigmoid. Eighty-two percent of patients had symptoms on presentation. CONCLUSIONS: Elective colorectal cancer surgery in Vermont is reasonably safe, and adherence to national standards for the use of adjuvant therapy is outstanding. Surgeons will provide outcome data if confidentiality and peer review protection are provided. The predominance of right-sided lesions and the low incidence of asymptomatic detection have significant implications for screening efforts in Vermont. PMID- 11926943 TI - Hand-assisted laparoscopic liver surgery. AB - HYPOTHESIS: The hand-assisted laparoscopic technique may be applied to the treatment of liver tumors. DESIGN: A case series with mean follow-up of 13 months. SETTING: University-affiliated tertiary care center. PATIENTS: A total of 15 patients with hepatic neoplasms underwent screening tests, including appropriate tumor marker analyses, abdominal sonography, and computed tomographic scan and, in most cases, magnetic resonance imaging to determine operability. Contraindications included extrahepatic disease, more than 5 liver lesions, coagulopathy, and ascites. INTERVENTION: Between March 1, 1998, and April 30, 2001, 15 patients underwent 16 hand-assisted diagnostic laparoscopic operations to rule out extrahepatic disease. Four patients had extrahepatic disease. In the 11 patients without evidence of extrahepatic disease, intraoperative ultrasound was used to establish the number and location of liver lesions. Operative strategies included resection, cryoablation, or both. MAIN OUTCOME MEASURES: Operative time, conversion to open procedure, length of stay, complications, and recurrence of disease. RESULTS: Of the 15 patients with liver tumors, 6 patients had more extensive disease than was detected by either preoperative imaging or laparoscopic exploration They included extrahepatic disease (3), additional liver lesion (2), or both (1). Hand-assisted management included resection only (3), cryoablation only (5), and a combination of the 2 (3). A total of 9 lesions were resected and 10 lesions were cryoablated. The mean operative time was 197 minutes with a mean length of stay of 4.5 days. There were no conversions to open procedures. One patient experienced minor postoperative bleeding but required no treatment. All treated patients are alive, and 5 have had recurrence of disease. CONCLUSIONS: Hand-assisted technique can be applied safely and effectively to laparoscopic liver surgery and may identify presence of otherwise undetectable disease. PMID- 11926945 TI - Lack of association of diabetes with increased postoperative mortality and cardiac morbidity: results of 6565 major vascular operations. AB - HYPOTHESIS: A number of preoperative factors, including diabetes mellitus (DM), have been cited as increasing risk in patients undergoing major vascular operations. In smaller studies at our institution we have not found this to be apparent. This study reviewed all major vascular operations to confirm our bias that DM is not associated with increased mortality or cardiac morbidity. DESIGN: Case series retrospectively reviewed from a vascular registry established in 1990. SETTING: Tertiary care center. PATIENTS: Consecutive sample of 6565 patients who underwent lower extremity revascularization or carotid or aortic procedures, grouped by presence or absence of DM. MAIN OUTCOME MEASURES: Postoperative mortality, congestive heart failure, or myocardial infarction. RESULTS: Patients with DM made up 62.3% of the population, and those without diabetes, 37.7%. Average age of the DM group was 67.1 years, with 61.3% male and 38.7% female. Average age of the non-DM group was 70.6 years, with 61.8% male and 38.2% female. The rates of overall postoperative mortality, myocardial infarction, and congestive heart failure were 1.14%, 1.59%, and 1.13%, respectively. Comparing the DM with the non-DM group, these rates were 0.96% vs 1.46%, 1.77% vs 1.30%, and 1.13% vs 1.14%, respectively. Using multivariate analysis, the DM group had an inverse relationship to perioperative death, with an odds ratio of 0.53 (P =.01). The factors that were associated with increased mortality were hemodialysis and history of congestive heart failure. Previous myocardial infarction was the only factor that predicted postoperative myocardial infarction. Kaplan-Meier survival curves showed a significantly decreased survival in the DM group during the next 5 years (P<.001). CONCLUSIONS: Diabetes alone does not confer a higher mortality or cardiac morbidity rate with major vascular procedures. However, long-term survival is significantly worse in this group of patients. PMID- 11926946 TI - Hepatic radiofrequency ablation. AB - HYPOTHESIS: Hepatic radiofrequency ablation (RFA) is effective in treating patients with unresectable hepatic malignancies. DESIGN: Case series of 123 patients with unresectable hepatic tumors or tumors with histological findings not traditionally treated by means of hepatic resection were considered for hepatic RFA. Median follow-up was 20 months. SETTING: Tertiary referral center. PATIENTS: The 123 patents underwent 168 RFA sessions from January 1, 1998, through September 30, 2001. Sixty-nine patients were male and 54, female; average age was 65 years (range, 1-89 years). Fifty-two patients had metastatic colorectal cancer; 30, hepatocellular carcinoma; and 41, cancers with other histological findings. INTERVENTIONS: A 200-W, cooled-tip RF probe system was used for all cases. Probe placement and ablation were monitored by means of real time ultrasonography or fluoroscopic computed tomography. Final tissue temperature of greater than 50 degrees C was achieved in all cases. RESULTS: Initial treatment sessions were percutaneous in 87 patients, open operations in 33, and laparoscopic in 3. Repeated sessions were percutaneous in all but 2 patients. The mean number of lesions treated per session was 2.7 (range, 1-24). Mean tumor size was 5.2 cm (range, 0.5-15.0 cm). One death occurred within 30 days of a procedure. No hepatic bleeds, bile leaks, or adult respiratory distress syndrome occurred. Overall morbidity was 7.1%. Complications included hepatic abscesses in 4 patients, transient liver insufficiency in 3, segmental hepatic infarcts in 2, diaphragm paralysis in 1, hepatic artery-to-portal vein fistula in 1, and systemic hemolysis in 1. CONCLUSIONS: Hepatic RFA is an effective treatment option for patients with unresectable hepatic malignancies. Careful patient selection based on tumor size, location, and number and on patient clinical status should determine the choice of treatment. Further controlled trials are needed to determine the effect of hepatic RFA on long-term survival. PMID- 11926947 TI - The effect of comorbid illness on mortality outcomes in cardiac surgery. AB - HYPOTHESIS: Comorbid conditions are associated with the risk of death from coronary artery bypass graft surgery. DESIGN: Prospective cohort study data were collected on patient and disease characteristics and comorbid conditions including hypertension, diabetes, obesity, vascular disease, chronic obstructive pulmonary disease, cancer (excluding nonmelanoma skin cancer), dialysis-dependent renal failure, liver disease, and dementia. Statistical analysis used logistic regression for the calculation of adjusted odds ratios (ORs) and 95% confidence intervals (CIs). SETTING: Regional cardiac surgery database. PATIENTS: A total of 27,239 consecutive patients undergoing isolated coronary artery bypass graft surgery. MAIN OUTCOME MEASURE: In-hospital mortality rate. RESULTS: The prevalence of comorbid conditions was as follows: hypertension, 64.3%; diabetes, 30.1%; obesity, 24.6%; severe obesity, 7.2%; vascular disease, 18.3%; chronic obstructive pulmonary disease, 10.9%; peptic ulcer, 7.5%; cancer, 3.8%; renal failure, 1.5%; liver disease, 0.6%; and dementia, 0.1%. After adjustment for patient and disease characteristics, including age, sex, previous cardiac surgery, priority of surgery, degree of left main coronary stenosis, number of diseased coronary arteries, and left ventricular ejection fraction, the following comorbid conditions were significant predictors of in-hospital mortality: diabetes (OR, 1.19; 95% CI, 1.01-1.40; P =.03), vascular disease (OR, 1.67; 95% CI, 1.41-1.97; P<.001), chronic obstructive pulmonary disease (OR, 1.57; 95% CI, 1.29-1.91; P<.001), peptic ulcer (OR, 1.34; 95% CI, 1.05-1.71; P =.02), and dialysis-dependent renal failure (OR, 3.68; 95% CI, 2.65-5.13; P<.001). There was no significant association between in-hospital mortality and hypertension, obesity or severe obesity, cancer, liver disease, or dementia. CONCLUSION: Even after adjustment for other patient and disease characteristics, comorbid conditions (especially diabetes, vascular disease, chronic obstructive pulmonary disease, peptic ulcer disease, and dialysis-dependent renal failure) are associated with significantly increased risk of death after coronary artery bypass graft surgery. PMID- 11926948 TI - Operative technique, paraplegia, and mortality after blunt traumatic aortic injury. AB - HYPOTHESIS: The use of mechanical circulatory support (MCS) during repair of traumatic aortic injuries is associated with a decreased incidence of postoperative paraplegia and mortality. DESIGN AND SETTING: Historical cohort study with contemporaneous but nonrandomized controls in a tertiary care hospital from July 1, 1988, through December 31, 1999. PATIENTS AND INTERVENTIONS: Consecutive cases undergoing operation for traumatic aortic injuries. Use of MCS (with or without systemic heparinization) determined by surgeon preference. MAIN OUTCOME MEASURES: Incidence of postoperative paraplegia and mortality. RESULTS: Twenty-two patients underwent repair of traumatic aortic injuries using MCS, resulting in no paraplegia but 4 deaths, 3 of them from cerebral ischemia. Thirteen patients had their traumatic aortic injuries repaired using a "clamp-and sew" or passive shunt technique with no deaths but paraplegia in 2. Compared with an earlier report from our group from January 1, 1975, through June 30, 1988, the annual incidence of traumatic aortic injuries has decreased, whereas the age of patients and proportion of operations using MCS have increased. A review of the recent literature on traumatic aortic injuries reveals an average postoperative paraplegia incidence of 1% with MCS and 16% without MCS. Overall mortality is similar, but others have also reported cases of cerebral ischemia after aortic repair. CONCLUSIONS: The use of MCS during repair of traumatic aortic injuries is associated with a decreased incidence of postoperative paraplegia. The occasional occurrence of cerebral ischemia deserves further study. PMID- 11926949 TI - Abnormal motility in patients with ulcerative colitis: the role of inflammatory cytokines. AB - HYPOTHESIS: Interleukin 1 beta (IL-1 beta) levels are elevated in the colonic mucosa of patients with ulcerative colitis (UC). We propose that IL-1 beta may also be elevated in the circular muscle layer of the colon and may be partially responsible for the motility dysfunction observed in patients with UC. DESIGN: Cohort analytic study. SETTING: Research laboratory in a tertiary academic medical center. PARTICIPANTS: Normal smooth muscle was obtained from the disease free margins of human sigmoid colon specimens resected from patients with cancer and compared with specimens from patients with UC. INTERVENTIONS: An enzyme linked immunosorbent assay was used to measure IL-l beta. Standard muscle chambers were used to measure force changes. Single muscle cells were isolated by enzymatic digestion, and cell shortening in response to neurokinin A (NKA) and thapsigargin was measured under a microscope. Cytosolic Ca(2+) (calcium) concentrations were measured by standard techniques. MAIN OUTCOME MEASURE: Effects of IL-1 beta on smooth muscle function in normal and UC colons. RESULTS: In patients with UC, IL-1 beta was elevated in the muscularis propria, and sigmoid circular smooth muscle contractions in response to NKA and thapsigargin were significantly reduced. In fura-2-loaded cells from patients with UC, the NKA induced Ca(2+) signal was also significantly reduced in Ca(2+)-free medium, indicating the reduced intracellular Ca(2+) stores after UC. Exposure of normal cells to IL-1 beta mimicked the changes observed in patients with UC. An IL-1 beta-induced reduction in contraction and release of intracellular Ca(2+) in response to NKA was partially restored by the hydrogen peroxide scavenger catalase. CONCLUSION: In patients with UC, IL-1 beta was increased in colonic circular muscles and may contribute to motor dysfunction after UC through production of hydrogen peroxide. PMID- 11926950 TI - Preoperative staging of rectal cancer using magnetic resonance imaging with external phase-arrayed coils. AB - HYPOTHESIS: Rectal cancer can be accurately staged preoperatively by magnetic resonance imaging (MRI) with external phase-arrayed coils. DESIGN: Comparison of MRIs with pathologic staging. SETTING: University hospital. PATIENTS: Twenty eight consecutive patients with biopsy-proven rectal cancer who did not undergo irradiation. INTERVENTION: Patients underwent imaging using a 1.5-T MRI scanner with external phase-arrayed surface coils. Streaking of the perirectal fat and disruption of the bowel wall margin were interpreted as transmural invasion. Lymph nodes were defined as metastatic when they had a diameter of at least 0.5 cm. Tumors were staged according to the TNM staging system (American Joint Committee on Cancer guidelines) as confined to the bowel wall (T1-T2) and invading through the bowel wall (T3-T4). Patients underwent anterior resection (n = 15), abdominoperineal resection (n = 11), or local excision (n = 2). MAIN OUTCOME MEASURES: Calculation of sensitivity, specificity, and accuracy for invasion through the bowel wall and lymph node status. RESULTS: Sensitivity of MRI in detecting invasion through the bowel wall was 89% (16/18), specificity was 80% (8/10), and accuracy was 86% (24/28). Sensitivity for malignant lymphadenopathy was 67% (8/12), specificity was 71% (10/14), and accuracy 69% (18/26). CONCLUSION: Although more costly and not as accurate as endoscopic ultrasound, MRI with phase-arrayed coils had excellent sensitivity at detecting transmural penetration of rectal cancer. PMID- 11926951 TI - The use of readily available equipment in a simple method for intraoperative monitoring of recurrent laryngeal nerve function during thyroid surgery: initial experience with more than 300 cases. AB - HYPOTHESIS: An inexpensive and widely applicable technique to monitor recurrent laryngeal nerve (RLN) function during thyroid surgery can be safely implemented. DESIGN: Consecutive sample. SETTING: Single-surgeon academic practice. PATIENTS: Three hundred sixty-three consecutive patients undergoing surgery for thyroid disease in the 18 months from November 1, 1999, to May 31, 2001. INTERVENTIONS: Anesthetic management using laryngeal mask airway and spontaneous ventilation, combined with electrical RLN stimulation and fiberoptic video laryngoscopy to confirm vocal cord response. MAIN OUTCOME MEASURES: Vocal cord response to RLN stimulation, intraoperative anatomic findings, postoperative voice quality, and anesthetic complications. RESULTS: We used the laryngeal mask airway-based technique in 327 cases. Visualization of vocal cords was maintained throughout the surgery in 310 cases (95%). In 10 cases (3%), the branch of the RLN carrying vocal cord function could not be identified until electrical testing was performed. A single inadvertent RLN palsy was seen in 1 of the 17 cases in which vocal cord visualization was lost during the procedure (0.03% overall). Upper airway obstruction occurred in 16 (5%) of 327 attempted procedures, requiring tracheal intubation in 3 (1%). No further complications regarding airway management were seen. Pneumothorax was observed in 5 cases (2%), each of which resolved without chest tube placement. CONCLUSIONS: This technique can be applied to thyroid surgery as a safe means of managing the airway. It is associated with an ability to test RLN function at will in more than 95% of cases using readily available equipment. PMID- 11926952 TI - Elective lymph node dissection in patients with melanoma: systematic review and meta-analysis of randomized controlled trials. AB - HYPOTHESIS: Elective lymph node dissection does not improve survival in patients with melanoma without clinically detectable lymph node metastases. OBJECTIVE: To determine whether elective lymph node dissection in patients with melanoma without clinically detectable regional metastases decreases overall mortality. DESIGN: Systematic review and meta-analysis of randomized controlled trials comparing elective lymph node dissection with delayed lymphadenectomy at the time of clinical recurrence. SETTING: Randomized controlled trials available by February 2001. SUBJECTS: The included trials comprised 1533 participants. INTERVENTION: Elective lymph node dissection compared with delayed lymphadenectomy or no lymphadenectomy in patients with melanoma without clinically detectable regional metastases. MAIN OUTCOME MEASURE: Overall mortality in treatment groups as compared with control groups at the end of a 5 year follow-up period. RESULTS: Three randomized controlled trials met the inclusion criteria. The pooled odds ratio for overall mortality for the 3 trials was 0.86 (95% confidence interval, 0.68-1.09). Results are statistically nonsignificant, but they have potential clinical significance. CONCLUSIONS: This systematic review of randomized controlled trials comparing elective lymph node dissection with surgery delayed until the time of clinical recurrence shows no significant overall survival benefit for patients undergoing elective lymph node dissection. Trials included in this review, however, contain significant bias. The question is not answered for all patients, and the results do not exclude the possibility that some subgroups may benefit from elective lymph node dissection. Further research is required. PMID- 11926954 TI - Ultrasound-guided thrombin injection is the treatment of choice for femoral pseudoaneurysms. AB - HYPOTHESIS: Ultrasound-guided injection of thrombin is the treatment of choice for femoral pseudoaneurysms. DESIGN: A prospective study of patients undergoing a single form of treatment for femoral pseudoaneurysms. SETTING: Patients were treated at 2 tertiary care, university-affiliated hospitals. PATIENTS: Forty consecutive patients with femoral pseudoaneurysms resulting from diagnostic and therapeutic catheterizations for peripheral arterial, coronary arterial, and cerebrovascular diseases were evaluated. INTERVENTION: Thrombin was injected directly into the pseudoaneurysms under ultrasonographic guidance. MAIN OUTCOME MEASURE: Thrombosis of the pseudoaneurysm following thrombin injection was the goal of treatment. RESULTS: All 40 patients had initial complete thrombosis of their femoral pseudoaneurysms. In each case, except for one, the aneurysm was noted to have thrombosed on follow-up duplex ultrasonogram. There was one complication. CONCLUSION: Ultrasound-guided thrombin injection is the treatment of choice for femoral pseudoaneurysms. PMID- 11926955 TI - Choledochal cysts in adults. AB - HYPOTHESIS: Choledochal cyst is rarely diagnosed in adulthood. When complicated by biliary tract malignancy, the disease has a distinct presentation and carries a dismal prognosis despite radical surgical resection. DESIGN: Retrospective study. SETTING: Tertiary referral center. PATIENTS: A retrospective study was performed on 30 adult patients who presented with choledochal cyst from January 1, 1989, to December 31, 2000. MAIN OUTCOME MEASURES: The clinical presentation, management, and outcome of patients with and without biliary tract malignancy. RESULTS: Nine patients (30%) had biliary tract malignancy complicating choledochal cyst (group A). Compared with 21 patients without malignancy (group B), group A patients had a significantly higher incidence of previous internal drainage operations for choledochal cyst (P =.049) and presentation with cholangitis (P =.03). Four patients in group A underwent pancreaticoduodenectomy and 3 received a palliative biliary drainage operation. The overall median survival of patients in group A was 12 months. Complete excision of choledochal cyst and Roux-en-Y hepaticojejunostomy were performed for all patients in group B, among whom 2 underwent concomitant hemihepatectomy. The operative morbidity and mortality were 14% and 0%, respectively, and there were no long-term complications with a median follow-up of 66 months. CONCLUSIONS: Biliary tract malignancy complicating choledochal cyst in adults should be suspected in patients with a history of internal drainage of choledochal cyst and presentation with cholangitis. Complete excision of choledochal cyst with Roux-en-Y hepaticojejunostomy is the treatment of choice for patients without malignancy and can be performed with low operative morbidity and absence of long-term complications in adult patients. PMID- 11926957 TI - Intraoperative lavage cytologic analysis of surgical margins as a predictor of local recurrence in pulmonary metastasectomy. AB - HYPOTHESIS: Cytologic analysis of intraoperative lavage at the surgical margin during wedge or segmental resection for pulmonary metastatic lesions predicts postoperative local failure at the surgical margin of the pulmonary parenchyma. DESIGN: Prospective nonrandomized trial. SETTINGS: Institution-based study. PATIENTS: Fifty-one consecutive patients undergoing wedge or segmental resection for 87 pulmonary metastatic lesions of various primary tumor types from November 1, 1997, through January 31, 2001, were prospectively enrolled. INTERVENTIONS: An intraoperative lavage cytologic technique at the surgical margin for each pulmonary metastasis was performed as described previously. MAIN OUTCOME MEASURES: Incidence of positive cytologic findings and postoperative local recurrence at the surgical margin. RESULTS: Of the examined lesions, 10 (11%) showed positive cytologic results at the surgical margin, despite a macroscopically safe margin in the attempted resection. Of these, metastasectomy was converted to segmentectomy in 3. An additional wedge resection and evaporation using an Nd:YAG laser in the surgical margin were performed in 1 and 4 lesions, respectively. Complications precluded further treatment in 2 lesions. By July 2001, although no local recurrence at the surgical margin area was found among the lesions with negative cytologic results, recurrence at the surgical margin occurred in 2 with positive cytologic results, including 1 receiving no treatment and 1 receiving Nd:YAG laser evaporation, indicating that a significant difference in the recurrence rate according to lavage cytologic status (P<.001). CONCLUSIONS: This intraoperative lavage cytologic technique in wedge or segmental resection of pulmonary metastases of various primary tumors may be a useful predictor of local recurrence at the surgical margin. With these test results, local recurrence at the surgical margin may be controllable in patients undergoing pulmonary metastasectomy. PMID- 11926958 TI - Prognostic value of cytologic examination of peritoneal washings in pancreatic cancer. AB - HYPOTHESIS: Findings from cytologic examination of peritoneal washings affect the staging, prognosis, and management of pancreatic cancer. DESIGN: Retrospective review. SETTING: Tertiary care center. PATIENTS: A total of 151 patients with invasive ductal adenocarcinoma of the pancreas and without distant metastases or ascites, as determined by preoperative radiologic examinations, underwent peritoneal washings. Based on intraoperative findings, patients were subdivided into 3 groups according to the extent of cancer spread: 65 were assigned to a resectable group, 53 to a locally advanced group, and 33 to a visible metastases group. MAIN OUTCOME MEASURES: Survival and peritoneal metastases-free survival. RESULTS: The incidence of positive cytologic findings was 23.8% (36/151). Positive rates increased significantly with disease progression (P<.001). In the resectable group, survival and peritoneal metastases-free survival were significantly shorter for patients with positive vs negative cytologic findings. In the remaining 2 groups, no differences were observed between patients with positive vs negative cytologic findings in survival or peritoneal metastases-free survival. In patients with positive cytologic findings without visible metastases, survival and peritoneal metastases-free survival were significantly better in the locally advanced group undergoing chemoradiotherapy than in the resectable group. CONCLUSIONS: Positive peritoneal cytologic findings are not independent factors that determine survival and peritoneal metastases-free survival; rather, they are associated with advanced disease. In patients with visible metastases, cytologic factors are not correlated with survival or peritoneal recurrence. In patients without visible metastases, chemoradiotherapy may be beneficial for those with positive cytologic findings. PMID- 11926959 TI - Estimation of surgical costs using a prediction scoring system: estimation of physiologic ability and surgical stress. AB - HYPOTHESIS: Our predictive scoring system, Estimation of Physiologic Ability and Surgical Stress, can estimate surgical costs. DESIGN: Multicenter cohort study for 1 year. SETTING: Six national hospitals in Japan. PATIENTS: A consecutive series of 929 patients who underwent elective gastrointestinal operations. MAIN OUTCOME MEASURES: The preoperative and the comprehensive risk scores of the Estimation of Physiologic Ability and Surgical Stress were determined preoperatively and immediately after the operation, respectively. Estimated costs were computed using the following equation: costs = US $10,160 + (US $13,470 x comprehensive risk score). Data on length of stay, costs for surgical admission, and severity of postoperative complications were collected at hospital discharge. RESULTS: The comprehensive risk score significantly correlated with the severity of the postoperative complications (Spearman rank correlation = 0.54, P<.001), the length of stay (Spearman rank correlation = 0.69, P<.001), and the costs (Spearman rank correlation = 0.72, P<.001). The ratio of real to estimated costs varied from 0.82 to 1.17 at the various ranges of the comprehensive risk score, resulting in 0.93 in the total 929 patients. This ratio varied from 0.71 to 1.12 among the hospitals, the smallest of which was attributed to the hospital that primarily used the clinical pathways. A significant increase in the costs was observed according to the preoperative risk score for open colectomy (P =.009) and distal gastrectomy (P =.002). When we simulated the hospital revenue where different payment rates were set according to the preoperative risk score, the revenue seemed to improve in the hospitals that treated more high-risk patients, compared with the fixed payment system. CONCLUSION: The Estimation of Physiologic Ability and Surgical Stress scoring system may be useful for estimating surgical costs, making a benchmark analysis, and determining the rate in a risk-based payment system. PMID- 11926960 TI - It's acronymania all over again: with due reference to YB Yogi Berra. PMID- 11926962 TI - Image of the month. Acute respiratory distress syndrome. PMID- 11926963 TI - The validity of unplanned returns to the operating room as an indicator of quality of hospital care. PMID- 11926965 TI - Elements of surgery: America's first systematic treatise on surgery. PMID- 11926966 TI - An MCASE approach to the search of a cure for Parkinson's Disease. AB - BACKGROUND: Parkinson's disease is caused by a dopamine deficiency state in the fore brain area. Dopamine receptor agonists, MAO-B inhibitors, and N-Methyl-D Aspartate (NMDA) receptor antagonists are known to have antiparkinson effect. Levodopa, a dopamine structural analog, is the best currently available medication for the treatment of Parkinsons disease. Unfortunately, it also induces side effects upon long administration time. Thus, multidrug therapy is often used, in which various adjuvants alleviate side effects of levodopa and enhance its antiparkinsonian action. RESULTS: Computer models have been created for three known antiparkinson mechanisms using the MCASE methodology. New drugs for Parkinsons disease can be designed on the basis of these models. We also speculate that the presence of biophores belonging to different groups can be beneficial and designed some potential drugs along this line. The proposed compounds bear pharmacophores of MAO-B inhibitors, dopamine agonists and NMDA antagonists, which could synergistically enhance their antiparkinson effect. CONCLUSIONS: The methodology could readily be expanded to other endpoints where drugs with multiple activity mechanisms would be desirable. PMID- 11926967 TI - Down-regulation of the alpha-2C adrenergic receptor: involvement of a serine/threonine motif in the third cytoplasmic loop. AB - BACKGROUND: The mechanisms by which alpha-2 adrenergic receptors are down regulated following chronic exposure to agonist are not well understood. Interestingly, the human alpha-2C receptor does not down-regulate, whereas the opossum alpha-2C receptor does down-regulate. A comparison of the amino acid sequence of the third intracellular loop of these two receptors shows that the opossum alpha-2C receptor contains a potential G protein-coupled receptor kinase (GRK)phosphorylation motif (EESSTSE) with four hydroxyl residues, whereas the human alpha-2C receptor motif only contains two hydroxyl residues (DESSAAAAE). Because a similar acidic serine-rich motif (EESSSSD) in the human alpha-2 adrenergic receptor has been demonstrated to be phosphorylated by GRK and all four serines are required for desensitization of the receptor, we sought to determine whether the EESSTSE sequence was involved in the down-regulation of the alpha-2C adrenergic receptor. RESULTS: Site-directed mutagenesis was used to mutate the opossum alpha-2C receptor to SSVA and AAVA in place of the SSTS wild type sequence. Down-regulation experiments on CHO cells transfected with the receptors demonstrated that neither of the mutated receptors down-regulated following 24 h exposure to norepinephrine, whereas the wild-type receptor down regulated to 65 +/- 10% of the control. CONCLUSIONS: These results indicate that a motif with four hydroxyl amino acid residues in an acidic environment is important for down-regulation of the opossum alpha-2C adrenergic receptor. Because these are potential GRK phosphorylation sites, we suggest that GRK phosphorylation may be involved in alpha-2C adrenergic receptor down-regulation. PMID- 11926968 TI - High-susceptibility of photosynthesis to photoinhibition in the tropical plant Ficus microcarpa L. f. cv. Golden Leaves. AB - BACKGROUND: The tropical plant Ficus microcarpa L. f. cv. Golden Leaves (GL) is a high-light sensitive tropical fig tree in which sun-leaves are yellow and shade leaves are green. We compared the response of photosynthetic activities to strong light between GL and its wild-type (WT, Ficus microcarpa L. f.). RESULTS: Field measurements of maximum photosystem II (PSII) efficiency (Fv/Fm) of intact sun leaves in GL showed that photo synthetic activity was severely photoinhibited during the daytime (Fv/Fm = 0.46) and subsequently recovered in the evening (Fv/Fm = 0.76). In contrast, WT did not show any substantial changes of Fv/Fm values throughout the day (between 0.82 and 0.78). Light dependency of the CO2 assimilation rate in detached shade-leaves of GL showed a response similar to that in WT, suggesting no substantial difference in photosynthetic performance between them. Several indicators of photoinhibition, including declines in PSII reaction center protein (D1) content, Fv/Fm value, and O2 evolution and CO2 assimilation rates, all indicated that GL is much more susceptible to photoinhibition than WT. Kinetics of PAM chlorophyll a fluorescence revealed that nonphotochemical quenching (NPQ) capacity of GL was lower than that of WT. CONCLUSION: We conclude that the photosynthetic apparatus of GL is more highly susceptible to photoinhibition than that of WT. PMID- 11926969 TI - Diastolic blood pressure-estimated left ventricular dp/dt. AB - BACKGROUND: Peak dp/dt is one of the best isovolumic phase indexes of the myocardial contractile state requiring invasive procedures or presence of mitral regurgitation severe enough to measure in clinical practice by Doppler echocardiography. In this study, we sought the correlation between two noninvasive methods of measurements for left ventricular dp/dt-diastolic blood pressure- (DBP) estimated and continuous-wave Doppler-derived dp/dt-min electrocardiographic/echocardiographic study to emphasize the clinical feasibility of the DBP-estimated method. METHOD: Thirty-six randomized patients (27 male, 9 female; 58 +/- 8 years) with mild mitral regurgitation were enrolled in this study. DBP-estimated dp/dt was calculated from DBP minus the left ventricular end-diastolic pressure (LVEDP) over the isovolumetric contraction time (IVCT). LVEDP was assumed to be 10 mmHg for all patients. Doppler-determined left ventricular dp/dt was derived from the continuous-wave Doppler spectrum of mitral regurgitation jet by dividing the magnitude of the left ventricular atrial pressure gradient rise between 1 mm/sec-3 mm/sec of mitral regurgitant velocity signal by the time taken for this change. RESULTS: Left ventricular dp/dt by Doppler was 1122 +/- 303 mmHg/sec and blood pressure-estimated dp/dt was 1063 +/- 294 mmHg/sec. There was a high correlation (r = 0.97, P < 0.001) of dp/dt between the two techniques. CONCLUSIONS: DBP and IVCT can generate left ventricular dp/dt without invasive procedures, even in the absence of mitral regurgitation in clinical practice. PMID- 11926970 TI - Noninvasive estimation of pulmonary capillary wedge pressure by color M-mode Doppler echocardiography in patients with acute myocardial infarction. AB - The peak early diastolic filling velocity/flow propagation velocity (E/FPV) by color M-mode Doppler provides a better estimate of pulmonary capillary wedge pressure (PCWP) than transmitral or pulmonary venous flow. However, the value of E/FPV for the assessment of PCWP has not been evaluated in patients with acute myocardial infarction. We investigated the correlation between E/FPV and PCWP and whether PCWP can be estimated from E/FPV in patients with acute myocardial infarction. One hundred and two patients with acute myocardial infarction were divided into two groups. The first 60 patients were used to generate an equation to estimate PCWP (retrospective group). This equation was then assessed prospectively in the remaining 42 patients (prospective group). We measured the transmitral flow velocity indices and the deceleration time of diastolic pulmonary venous flow and E/FPV by Doppler echocardiography and compared these variables with PCWP measured using a pulmonary artery catheter. E/FPV was strongly correlated with PCWP (r = 0.89) in the retrospective group. The sensitivity of an E/FPV of > or = 2.0 for predicting a PCWP of > or = 18 mmHg was 95%, and the specificity was 98%. The estimated PCWP showed a strong correlation with the measured PCWP (r = 0.84, P < 0.0001) in the prospective group. The mean difference between the measured and estimated PCWP was - 0.4 +/- 3.6 mmHg. In patients with acute myocardial infarction, E/FPV by color M-mode Doppler during early left ventricular filling provides a better estimate of PCWP than transmitral or pulmonary venous flow. PMID- 11926971 TI - False positive ST segment elevation during dobutamine stress echocardiography due to left ventricular hypertrophy. AB - The significance of ST segment elevation in dobutamine stress echocardiography (DSE) remains controversial. In patients with prior Q wave myocardial infarction (MI), it may reflect myocardial ischemia, contractile reserve in the infarct related area, or dyskinesia of the infarcted areas of myocardium. In the nonpost MI population, it has been attributed to vasospasm or strongly associated with coronary artery disease and ischemia. We hypothesized that ST segment elevation in the absence of inducible ischemia or prior MI is related to the presence of left ventricular hypertrophy (LVH). During DSE, dobutamine was infused from 5 microg/kg/min up to a maximum of 50 microg/kg/min. Echocardiographic images were obtained at baseline, low dose, peak dose, and recovery. Ischemia was defined as either the development of a new wall-motion abnormality or worsening wall motion at peak dose. We reviewed 682 consecutive DSE tests and found ST elevation in 42 patients (incidence = 6.1%). After excluding two patients for > 10% uninterpretable echocardiographic segments, the study population consisted of 40 patients. In 25 patients with ST elevation and without echocardiographic evidence for dobutamine-induced ischemia, 21 (84%) patients had LVH (P = 0.001). In 15 patients with inducible ischemia, only 4 (27%) patients had LVH. No other significant differences were found except that prior MI was more common in the inducible ischemia group. In the subgroup of 18 patients without prior MI, no inducible ischemia was found in 15 (83%). LVH was present in 14 (93%) of these 15 patients (P < 0.005), and all 14 had a normal baseline left ventricular ejection fraction. None of the three patients in the nonpost-MI subgroup with inducible ischemia had LVH. The 22 patients with prior MI had no significant association with LVH (P = 0.39). We conclude that ST segment elevation during DSE can occur without echocardiographic evidence for ischemia and is associated with LVH in the nonpost-MI population. This ST elevation may be related to transient electrocardiographic repolarization changes in the hypertrophied ventricle in the presence of altered loading conditions and/or altered catecholamine influences rather than true ischemia. PMID- 11926972 TI - Endocardial border delineation during dobutamine infusion using contrast echocardiography. AB - BACKGROUND: A significant percentage of pharmacologic stress echocardiograms produce suboptimal images despite the use of second harmonic imaging. Intravenous continuous infusion of myocardial ultrasound contrast may enhance endocardial border delineation during dobutamine-atropine stress echocardiography (DASE), improving wall-motion analysis. PATIENTS AND METHODS: We prospectively studied 68 patients (41 males and 27 females), mean age 58 years, with DASE during intravenous infusion of contrast using second harmonic imaging. Dobutamine was infused in scalar doses of 5 microg/kg/min to 40 microg/kg/min, and atropine was administered in doses of up to 1 mg. We diluted 0.1 mL of perfluorocarbon-exposed sonicated dextrose albumin (PESDA) microbubbles into 80 mL of saline solution, which was used for continuous intravenous infusion. Blinded reviewers used a 16 segment model at rest and peak DASE to analyze segmental wall delineation in two sets of images for each patient, with and without contrast. An endocardial delineation score of 0-3 (nondelineated to excellent delineation) was given to each segment. An endocardial delineation score index (EDSI), the number of endocardial delineation scores for each set of images divided by 16, was created. RESULTS: The analysis of the mean EDSI for the 2176 segments was 1.46 (+/- 0.43) at rest and 1.30 (+/- 0.48) at peak for noncontrast images and 2.22 (+/- 0.52) and 2.29 (+/- 0.52) for contrast images. Complete left ventricle opacification was obtained in all patients, with a mean dose of 4 mL/min, although in 15 (22%) patients, signs of apical bubble destruction occurred. There were 1768 (81%) of 2176 segments delineated without contrast enhancement and 2057 (95%) of 2176 with enhancement (P < 0.05). CONCLUSION: Continuous infusion of myocardial ultrasound contrast improves endocardial border delineation using second harmonic imaging in patients undergoing DASE. PMID- 11926973 TI - Effect of left ventricular global systolic function, mitral regurgitation, and left ventricular inflow pattern on exercise echocardiography results. AB - Diagnosis of coronary artery disease (CAD) by exercise echocardiography is usually based on rest or exercise-induced regional wall-motion abnormalities. Mitral regurgitation (MR), left ventricular (LV) global systolic function, and LV inflow measurements can be assessed during exercise echocardiography; however, their diagnostic value has not been analyzed consistently. Treadmill exercise echocardiography and coronary angiography were performed in 120 patients (94 male, 26 female; mean age 61 +/- 10 years [+/- 1 SD]) to evaluate known or suspected CAD. Positive exercise echocardiography was defined either as a rest- or exercise-induced regional wall-motion abnormalities. An abnormal response of LV ejection fraction (EF), LV volumes, MR (as assessed by color Doppler), and LV inflow pattern was defined as a fall in LVEF, a LV end-diastolic volume increase, a LV end-systolic volume increase, a new or increased MR, or a change from an impaired relaxation pattern (E < A) to a "pseudonormalized" pattern (E > A) from rest to exercise, respectively. CAD (> or = 50% luminal narrowing in at least one vessel) was found in 89 (74%) patients. EE-based regional wall-motion abnormality analysis was positive in 95 (79%) patients and negative in 25 (21%) patients. Feasible images for regional wall-motion abnormalities, LVEF and volumes, LV inflow, and MR measurements were acquired in 90% of patients. Regional wall motion abnormality analysis and LVEF decrease provided the greatest sensitivities for CAD (94% and 75%, respectively), while the highest specificity was given by a new or increased MR (90%), the development of a pseudonormalized pattern (88%), and the appearance of angina (87%). A positive electrocardiogram (ECG) finding in patients with interpretable ECGs provided good sensitivity and specificity (67% and 85%, respectively). In conclusion, a complete rest and exercise Doppler echocardiography approach is feasible in most patients. Regional wall-motion abnormalities are the most accurate exercise echocardiography variable for diagnosing CAD, whereas exercise ECG remains a good test in patients with interpretable ECGs. Exercise echocardiography, exercise ECG, newly developed or increased MR, and change to a pseudonormalized LV inflow pattern are highly specific. PMID- 11926974 TI - Does rapid volume loading during transesophageal echocardiography differentiate constrictive pericarditis from restrictive cardiomyopathy? AB - BACKGROUND: Respiratory variation of the pulmonary venous (PV) peak flow velocities can be used to distinguish constrictive pericarditis (constriction) from restrictive cardiomyopathy (restriction). Rapid volume expansion has been used successfully to enhance diastolic pressure equalization in occult constriction. The effect of volume on the respiratory variation in constriction has not been studied previously. This study assessed the utility of volume in enhancing the PV respiratory variation of constriction to further separate it from restriction. METHODS: The study population consisted of 15 patients referred to the echocardiography laboratory for further evaluation of clinically suspected diastolic dysfunction. Pulsed-Doppler transesophageal echocardiography (TEE) of the left or right upper pulmonary vein and mitral inflow was performed with respiratory monitoring before and after infusion of 1 liter of normal saline over 5 to 10 minutes. The classification of patients as constriction (n = 8) or restriction (n = 7) was confirmed independently by cardiac catheterization or surgery. Peak velocities of the PV systolic and diastolic waves and the mitral inflow E were measured during inspiration and expiration. A mean of 3-6 respiratory cycles was obtained for each value before and after volume loading. The percent change from expiration to inspiration (%E) was calculated using the formula %E = expiration - inspiration / expiration. RESULTS: At baseline, patients with constrictive pericarditis can be separated reliably from those with restrictive cardiomyopathy based on a higher systolic/diastolic ratio and greater respiratory variation of their PV diastolic flow velocity. There were no complications in any patient due to volume expansion. Although the change from baseline to volume expansion was not statistically significant in either constriction or restriction, the %E of the PV diastolic wave became significantly higher in constriction than in restriction (P < 0.05). CONCLUSIONS: Rapid volume expansion is relatively safe during TEE and can be used for further separation of constrictive pericarditis from restrictive cardiomyopathy by significantly enhancing the respiratory variation of the PV diastolic flow velocity in constrictive pericarditis. PMID- 11926975 TI - Free-hand four-dimensional echocardiography: a diagnostic tool in coronary artery disease. AB - Direct echocardiographic visualization of coronary arteries has been difficult due to the inherent problems involved in obtaining ultrasound access. We used freehand four-dimensional echocardiography in a 79-year-old patient with a trifurcational stenosis. We were able to identify a 6 x 11 mm possibly subintimal atheromathous plaque and to measure the diameters of the left descending coronary artery before, in, and behind the stenosis. There was good agreement with the angiographic measurements. This successful approach may be an argument to motivate further research in this area. PMID- 11926976 TI - Embolic cerebral infarction and gastrointestinal hemorrhage following thrombolytic therapy for acute myocardial infarction. AB - We report an unusual case of cerebral embolization that occurred after intravenous thrombolytic therapy for myocardial infarction. Direct observation by serial echocardiograms in this patient confirmed that the thrombolytic treatment induced lysis and fragmentation of thrombus, and the subsequent dislodging and embolization of preexisting cardiac thrombi, which caused the cerebral infarction. It is suggested that an echocardiogram, if instantly available, be performed before considering thrombolytic therapy whenever acute anterior wall myocardial infarction is impressed. PMID- 11926978 TI - Transesophageal echocardiographic evidence of atrial fibrillation with systolic flow in the right atrial appendage and systolic reversal pulmonary flow in a patient with DDD permanent pacing. AB - We report the case of a patient with atrial fibrillation on standard electrocardiogram who underwent transesophageal echocardiography to rule out presence of thrombi in heart cavities. Pulsed-wave Doppler investigation revealed multiphasic waves consistent with atrial fibrillation in the left atrial appendage, while in the right atrial appendage, a biphasic flow was registered after QRS depolarization. Some hypotheses are made about the origin of this signal. PMID- 11926977 TI - Anterolateral papillary muscle rupture: diagnosis and successful treatment (a case report). AB - We report the case of a young man presenting with chest pain, dyspnea, and syncope in whom transthoracic and transesophageal echocardiography helped to diagnose anterolateral papillary muscle rupture. After cardiac catheterization (which confirmed the severe mitral regurgitation and showed two vessel coronary disease), mitral valve replacement was performed together with coronary bypass grafting. PMID- 11926979 TI - Renal cell carcinoma presenting as right atrial mass. AB - Approximately 190 new cases of renal cell carcinoma infiltrate into the inferior vena cava and right atrium every year. Echocardiography often plays a major role in establishing the diagnosis and in guiding surgical therapy. This report describes a patient who presented with shortness of breath but had no classic signs or symptoms of renal cell carcinoma other than chronic anemia. Transthoracic two-dimensional echocardiography and transesophageal echocardiography established the diagnosis of a right atrial mass extending from the inferior vena cava that was later discovered to be renal cell carcinoma. PMID- 11926980 TI - Isolated noncompaction of the ventricular myocardium: contrast echocardiographic findings and review of the literature. AB - Isolated noncompaction of the ventricular myocardium (INVM) is a rare congenital anomaly. We report a case of INVM in an adult, which was diagnosed using contrast echocardiography. PMID- 11926981 TI - Proximal isovelocity surface area (PISA) in the evaluation of fixed membranous subaortic stenosis. AB - The evaluation of the severity of subaortic stenosis is usually expressed by the magnitude of the subvalvular gradient. Calculation of the membrane orifice area noninvasively is difficult by the standard means. We present a patient in whom the area was calculated using the proximal isovelocity surface area (PISA) method. This method should have clinical applicability because it is not flow dependent and can be used in patients with normal, reduced, or increased stroke volume. PMID- 11926982 TI - Anomalous origin of the right coronary artery from the left sinus of valsalva: diagnosis by multiplane transesophageal echocardiography. PMID- 11926983 TI - False tendon connecting the anterior mitral valve leaflet to the roof of the left atrium. PMID- 11926984 TI - Transesophageal three-dimensional echocardiographic assessment of left ventricular pseudoaneurysm. AB - We report an adult patient with a left ventricular pseudoaneurysm following an acute myocardial infarction in whom three-dimensional (3-D) transesophageal echocardiography (TEE) delineated clearly not only the location but also the size and shape of the rupture site. The size of the rupture site measured by 3-D TEE correlated well with the surgical measurements. Three-dimensional images also showed a localized superior distortion of the lateral aspect of the mitral annulus and left atrial wall produced by the pseudoaneurysm. The resulting severe mitral regurgitation practically disappeared after repair and decompression of the pseudoaneurysm. PMID- 11926985 TI - Phosphoinositide 3-kinases in immunity: lessons from knockout mice. AB - Phosphoinositide 3-kinases (PI3Ks) constitute a family of evolutionarily conserved lipid kinases that phosphorylate the D3 position of the inositol ring of phosphoinositides and produce PI(3)P, PI(3,4)P(2), and PI(3,4,5)P(3). Intense in vitro research over the last decade has unequivocally demonstrated that PI3Ks, in particular those belonging to class I, regulate a vast array of fundamental cellular responses. Given the pleiotropic roles of PI3Ks and the lipid product PI(3,4,5)P(3) in plethora of cellular responses, it is pertinent to explore the significance of PI3K signaling in vivo. In the past two or three years, the components of this signaling pathway have been genetically manipulated in mouse. This review briefly summarizes the immunological significance of PI3K signaling as revealed by the study of gene-targeted "knockout" mice. PMID- 11926986 TI - Phosphoinositide kinases as enzymes that produce versatile signaling lipids, phosphoinositides. AB - Phosphoinositide kinases comprise a unique family of enzymes that catalyze the phosphorylation of phosphatidylinositol and its phosphorylated metabolites to produce seven phosphoinositides. Recent advances have revealed that these phosphoinositides have specific physiological functions, such as in actin cytoskeletal reorganization, membrane transport, cell proliferation and survival, in eukaryotic cells and that each phosphoinositide kinase is differently and precisely regulated. Here we describe the diverse regulation and physiological functions of phosphoinositide kinases involving their products. PMID- 11926987 TI - N-glycans stabilize human erythropoietin through hydrophobic interactions with the hydrophobic protein surface: studies by surface plasmon resonance analysis. AB - Human erythropoietin (EPO) produced in Chinese hamster ovary cells is a hydrophobic protein highly stabilized by multibranched complex-type N-glycans. To reveal the molecular basis of the interaction between the N-glycans and the EPO protein, complex-type N-glycans of different structures were analyzed as to their binding affinity for Escherichia coli-expressed EPO by means of the surface plasmon resonance technique. It appears well established that complex-type N glycans, particularly multibranched ones, have hydrophobic regions that extensively stretch across the plane holding acetylamino groups and that N-glycan protein hydrophobic interactions characterized by a slow rate of dissociation stabilize the protein conformation. PMID- 11926988 TI - The possibility of reducing xenoantigen levels with a novel gal 3' sulfotransferase (GP3ST). AB - Glycoprotein-3-sulfotransferase (GP3ST) is a key enzyme in downregulating the expression of Galalpha1,3Galbeta1,4GlcNAc-R (the alpha-Gal epitope), via enzymatic competition with an alpha1,3 galactosyltransferase (alpha1,3GT), such as alpha2,6 sialyltransferase (alpha2,6ST). In this study, we report the dominance of GP3ST over alpha1,3GT using transfected pig endothelial cell (PEC) lines. The introduction of the GP3ST gene into PEC suppresses its antigenicity with respect to normal human pooled serum (NHS), including the alpha-Gal epitope and the Hanganutziu-Deicher (H-D) antigen, and, in addition, reduces the susceptibility to NHS in complement-mediated cell lysis. Western and lectin blot analyses of the products of parental PEC and its transfectants indicated that proteins smaller than 66 kDa have a diminished reactivity with NHS and the IB4 lectin. The levels of the alpha-Gal epitope in neutral glycosphingolipids were also decreased in the GP3ST transfectants as detected in thin layer chromatography by immunostaining. These data indicate that GP3ST is very effective in reducing xenoepitope levels. PMID- 11926989 TI - Accurate measurement of near-micromolar oxygen concentrations in aqueous solutions based on enzymatic extradiol cleavage of 4-chlorocatechol: applications to improved low-oxygen experimental systems and quantitative assessment of back diffusion of oxygen from the atmosphere. AB - An enzymatic method for measuring the O(2) concentrations of aqueous solutions was developed by involving 4-chlorocatechol and catechol 2,3-dioxygenase from Pseudomonas putida. With this system, the amount of O(2) in a sample solution can be measured as the amount of 5-chloro-2-hydroxymuconate semialdehyde formed through the enzyme reaction. The product was stable and its anion exhibited strong absorption around 380 nm (molar absorption coefficient of 4.3 x 10(4) M( 1) cm(-1), pK value of 5.4). A sensitive HPLC method involving a BioAssist Q column was developed to individually quantify the products derived from 4 chlorocatechol and catechol. When the O(2) concentration in a sample solution sealed in a vial was lowered from the air-saturation level by means of the amount enzymatically reacted with a known amount of catechol, the concentration of remaining O(2) could be successfully measured by the HPLC method. We developed devices through which reagents could be added to solutions sealed in cuvettes or the vessel of an oxygen electrode system under a flow of argon. By applying these devices, the submicromolar O(2) concentration of an anoxic solution and the back diffusion of O(2) from the atmosphere could be directly determined for the first time. The K(m) values of the dioxygenase and an ascorbate oxidase for oxygen were also determined to be 7.2 (at pH 7.5) and 114 microM (at pH 6.5), respectively, at 25 degrees C. PMID- 11926990 TI - PU.1 is dominant and HAF-1 supplementary for activation of the gp91(phox) promoter in human monocytic PLB-985 cells. AB - Gp91(phox) is a key component of the phagocyte NADPH oxidase. Mutations of its promoter found in patients with chronic granulomatous disease cause deficient binding of PU.1 and HAF-1. Because the two factors bind to the same site (Pu box) of the promoter, we attempted to clarify their relative in vivo contributions to activation of the gp91(phox) promoter in monocytically differentiated PLB-985 cells using a dual luciferase reporter assay and a gel shift competition assay. We found that the activity of a series of single-point-mutated promoters increases or decreases according to an increase or decrease, respectively, in the affinity of the promoters to PU.1 but not to HAF-1. Two of 7 mutants showing weak binding affinity to PU.1 exhibited moderate promoter activity and normal binding affinity for HAF-1. These results suggest PU.1 is the dominant activator and HAF 1 is supplementary. The increased promoter activity of single-, double-, and triple-point-mutated constructs with sequences closer to that of the Ets-binding element correlates with their binding affinity to PU.1 but not to HAF-1, supporting that PU.1 is a more efficient activator than HAF-1. In contrast to co expressed wild-type PU.1, dominant-negative PU.1 significantly inhibited the activity of a PU.1-optimised gp91(phox) promoter construct. Therefore, we conclude that PU.1 and HAF-1 binding to the Pu box is dominant and supplementary, respectively, for activation of the gp91(phox) promoter in human monocytic cells. PMID- 11926991 TI - Acidic phospholipids with unsaturated fatty acids inhibit the binding of origin recognition complex to origin DNA. AB - Origin Recognition Complex (ORC) is a candidate initiator of chromosomal DNA replication in eukaryotes. We recently reported that cardiolipin inhibits the interaction of Origin Recognition Complex ORC with origin DNA, as is the case of DnaA, the initiator of chromosomal DNA replication in prokaryotes. We report here that another acidic phospholipid, phosphatidylglycerol (PG), also inhibits the interaction. Synthetic PG with only unsaturated fatty acids inhibits ORC-binding to origin DNA more strongly than PG with only saturated fatty acids. On the other hand, phosphatidylcholine (neutral phospholipid) does not affect the ORC-origin interaction, regardless of the presence of saturated or unsaturated fatty acids. These results suggest that an acidic moiety and unsaturated fatty acids are important factors for the inhibitory effect of phospholipids on ORC binding to origin DNA, as is the case for DnaA. The inhibitory effect of cardiolipin on ORC binding to origin DNA was more apparent at 30 degrees C than at 4 degrees C. Furthermore, chlorpromazine restored the ORC-origin interaction in the presence of cardiolipin. Since the presence of unsaturated fatty acids, low incubation temperatures, and the addition of chlorpromazine all decrease membrane fluidity, these results suggest that membrane fluidity is important for the inhibitory effect of acidic phospholipids on ORC-binding to origin DNA, as is the case for DnaA. PMID- 11926992 TI - Macrophage binding and the uptake of oxidized low density lipoprotein are regulated by intracellular protein phosphorylation. AB - The involvement of intracellular protein phosphorylation in macrophages in the binding and uptake of oxidized low density lipoprotein (oxLDL) was investigated. The treatment of fibronectin-unstimulated and stimulated mouse thioglycolate induced macrophages with inhibitors of myosin light chain kinase, protein kinase C and protein tyrosine kinase resulted in decreased macrophage binding of oxLDL, macrophage foam cell formation, and whole intracellular protein phosphorylation. The treatment of fibronectin-unstimulated and stimulated macrophages with inhibitors of protein serine/threonine and tyrosine phosphatases caused enhanced macrophage binding of oxLDL, macrophage foam cell formation, and whole intracellular protein phosphorylation. Fibronectin, which stimulates macrophage activity, enhanced macrophage intracellular protein phosphorylation. Myosin light chain phosphorylation may be involved in the fibronectin stimulation of macrophages. Treatment of fibronectin-unstimulated and stimulated macrophages with thiophosphate, which forms thiophosphate esters of intracellular proteins that are not so susceptible to protein phosphatases, enhanced macrophage binding of oxLDL. The above results indicate that intracellular protein phosphorylation maintains and enhances macrophage binding and the uptake of oxLDL. PMID- 11926993 TI - Comparative study of the reaction mechanism of family 18 chitinases from plants and microbes. AB - Hydrolytic mechanisms of family 18 chitinases from rice (Oryza sativa L.) and Bacillus circulans WL-12 were comparatively studied by a combination of HPLC analysis of the reaction products and theoretical calculation of reaction time courses. All of the enzymes tested produced beta-anomers from chitin hexasaccharide [(GlcNAc)(6)], indicating that they catalyze the hydrolysis through a retaining mechanism. The rice chitinases hydrolyzed predominantly the fourth and fifth glycosidic linkages from the nonreducing end of (GlcNAc)(6), whereas B. circulans chitinase A1 hydrolyzed the second linkage from the nonreducing end. In addition, the Bacillus enzyme efficiently catalyzed transglycosylation, producing significant amounts of chitin oligomers larger than the initial substrate, but the rice chitinases did not. The time-courses of (GlcNAc)(6) degradation obtained by HPLC were analyzed by theoretical calculation, and the subsite structures of the rice chitinases were identified to be (-4)(-3)(-2)(-1)(+1)(+2). From the HPLC profile of the reaction products previously reported [Terwisscha van Scheltinga et al. (1995) Biochemistry 34, 15619-15623], family 18 chitinase from rubber tree (Hevea brasiliensis) was estimated to have the same type of subsite structure. Theoretical analysis of the reaction time-course for the Bacillus enzyme revealed that the enzyme has (-2)( 1) (+1)(+2)(+3)(+4)-type subsite structure, which is identical to that of fungal chitinase from Coccidioides immitis [Fukamizo et al. (2001) Biochemistry 40, 2448 2454]. The Bacillus enzyme also resembled the fungal chitinase in its transglycosylation activity. Minor structural differences between plant and microbial enzymes appear to result in such functional variations, even though all of these chitinases are classified into the identical family of glycosyl hydrolases. PMID- 11926994 TI - Paraquat- and diquat-induced oxygen radical generation and lipid peroxidation in rat brain microsomes. AB - NADPH-menadione reductase activity by rat brain microsomes (Ms) was decreased 40 50% by 10 microM dicumarol, a potent inhibitor of DT-diaphorase, whereas no change in NADPH-paraquat (PQ) and -diquat (DQ) reductase activity was observed. NADPH-DQ reductase activity in brain Ms was 2.5-fold higher than NADPH-PQ reductase activity. The formation of PQ and DQ radicals was verified optically and observed directly by ESR spectroscopy in the NADPH-PQ and -DQ reductase reactions by brain Ms under anaerobic conditions. PQ- and DQ-induced superoxide formation was confirmed by the detection of DMPO-OOH ESR signals and followed by chemiluminescence (CL) of a Cypridina luciferin analogue (CLA). The kinetics and intensity of the CL were consistent with the observations that the reduction in DQ is faster than that in PQ. Thiobarbituric acid reactive substances (TBARS) and phospholipid hydroperoxides in brain Ms increased in the presence of NADPH and Fe3+. The generation of both lipid peroxidation products derived from brain Ms decreased with increasing concentrations of PQ and DQ. The inhibitory effect of DQ is more pronounced than that of PQ. The formation of PQ- and DQ-induced reactive oxygen species was not associated with lipid peroxidation in rat brain Ms. PMID- 11926995 TI - Additivity of interactions of zinc finger motifs in specific recognition of RNA. AB - Xenopus transcription factor IIIA (TFIIIA) binds 5S rRNA and the 5S rRNA gene, these interactions being mediated by nine zinc fingers. To determine the contribution of each finger to the binding to 5S rRNA we prepared a series of peptides containing different numbers of zinc fingers and analyzed their interactions with RNA. The topography of these complexes was analyzed with the specific RNase and hydroxyl radical footprinting methods. Our results show the direct contribution of each zinc finger (ZF) peptide to the specific recognition of 5S rRNA. These data clearly suggest that total binding of TFIIIA with 5S rRNA is the sum of the specific interactions of the individual zinc fingers with RNA and that they have an additive character. PMID- 11926996 TI - Nitrate reductase-formate dehydrogenase couple involved in the fungal denitrification by Fusarium oxysporum. AB - Dissimilatory nitrate reductase (Nar) was solubilized and partially purified from the large particle (mitochondrial) fraction of the denitrifying fungus Fusarium oxysporum and characterized. Many lines of evidence showed that the membrane bound Nar is distinct from the soluble, assimilatory nitrate reductase. Further, the spectral and other properties of the fungal Nar were similar to those of dissimilatory Nars of Escherichia coli and denitrifying bacteria, which are comprised of a molybdoprotein, a cytochrome b, and an iron-sulfur protein. Formate-nitrate oxidoreductase activity was also detected in the mitochondrial fraction, which was shown to arise from the coupling of formate dehydrogenase (Fdh), Nar, and a ubiquinone/ubiquinol pool. This is the first report of the occurrence in a eukaryote of Fdh that is associated with the respiratory chain. The coupling with Fdh showed that the fungal Nar system is more similar to that involved in the nitrate respiration by Escherichia coli than that in the bacterial denitrifying system. Analyses of the mutant species of F. oxysporum that were defective in Nar and/or assimilatory nitrate reductase conclusively showed that Nar is essential for the fungal denitrification. PMID- 11926997 TI - Catalytic mechanism of beta-amylase from Bacillus cereus var. mycoides: chemical rescue of hydrolytic activity for a catalytic site mutant (Glu367-->Ala) by azide. AB - The hydrolytic activity of beta-amylase from Bacillus cereus var. mycoides was lost on replacement of either of the catalytic residues (Glu172 or Glu367) with an alanyl residue. When maltopentaose and 2 M azide existed together mutant, E367A cleaved the glucosidic linkage of maltopentaose and produced maltose at pH 7.0 and 25 degrees C, but the other mutants (E172A and double mutant E172A/E367A) did not. This indicates that azide acts as a general base instead of E367 and Glu172 acting as general acids, and that the hydroxide ion generated from a water molecule activated by azide attacks a reactive pyranose nucleophilically so that beta-maltose is produced. PMID- 11926998 TI - Inhibition of proteasomes induces accumulation, phosphorylation, and recruitment of HSP27 and alphaB-crystallin to aggresomes. AB - Molecular chaperones and the ubiquitin-proteasome pathway are known to participate in the quality control of proteins in cells. In this study, we examined the responses of small heat shock proteins to proteasome inhibitors to clarify their roles under conditions where misfolded proteins are abnormally accumulated. HSP27 and alphaB-crystallin accumulated in both soluble and, more prominently, insoluble fractions after exposure to MG-132, a proteasome inhibitor. Enhanced expression of mRNAs for HSP27 and alphaB-crystallin was observed, suggesting transcriptional activation. Phosphorylation of HSP27 and alphaB-crystallin in cells treated with MG-132 was enhanced concomitantly with activation of p38 and p44/42 MAP kinase pathways. Immunofluorescence analysis revealed that exposure to proteasome inhibitors induced the formation of aggresomes in U373 MG cells, to which HSP27 and alphaB-crystallin were recruited. However, phosphorylation was not required for this accumulation in aggresomes. Thus, HSP27 and alphaB-crystallin are increased, phosphorylated and localized in aggresomes when proteasome activity is inhibited. PMID- 11926999 TI - Identification of a domain conferring nucleotide binding to the N-acetyl-d glucosamine 2-epimerase (Renin binding protein). AB - Renin binding protein (RnBP), a cellular renin inhibitor, has been identified as the enzyme N-acetyl-D-glucosamine (GlcNAc) 2-epimerase. Our recent studies demonstrated that rat GlcNAc 2-epimerase has a ten-times higher affinity for ATP, dATP, and ddATP than the human enzyme [Takahashi, S. et al. (2001) J. Biochem. 130, 815-821]. To identify the domain conferring nucleotide binding to GlcNAc 2 epimerase, we constructed a series of chimeric enzymes successively replacing the three domains of the human enzyme (N-terminal, middle, and C-terminal domains) with the corresponding domains of the rat enzyme. Chimeras were expressed in Escherichia coli JM109 cells under the control of the Taq promoter. The purified chimeric enzymes had GlcNAc 2-epimerase activity and inhibited renin activity in a dose-dependent manner. The recombinant human and rat enzymes required catalytic amounts of ATP with apparent K(m) values of 73 and 5.5 microM, respectively. Chimeric enzymes of HHR, RHH, and RHR (H, human type domain; R, rat type domain) had nearly the same nucleotide specificity as the human GlcNAc 2-epimerase. On the other hand, HRR, HRH, and RRH chimeras had the same nucleotide specificity as the rat enzyme. These results indicate that the middle domain of the GlcNAc 2 epimerase molecule participates in the specificity for and binding of nucleotides, and that nucleotides are essential to form the catalytic domain of the enzyme. PMID- 11927000 TI - Characteristics of bovine hemoglobin as a potential source of hemoglobin-vesicles for an artificial oxygen carrier. AB - Hemoglobin-vesicles (HbV) have been developed for use as artificial O(2) carriers in which a purified Hb solution is encapsulated within a phospholipid bilayer membrane. In this study, bovine Hb (BHb) was tested as a source of HbV instead of human Hb (HHb). We compared the preparation process and characteristics of BHbV with those of HHbV. The purification of BHb was effectively performed simply with an ultrafiltration system including a process for removing virus and scrapie reagent. The removal ratio of the phospholipid components of bovine red blood cells was over 99.99%, and the protein purity was over 99.9%. The deoxygenated and carbonylated BHb showed denaturation transition temperatures at 83 and 87 degrees C, respectively, which are higher than those of HHb (80 and 78 degrees C, respectively), and resistant to pasteurization (60 degrees C, 10 h). The purified BHb was concentrated to over 40 g/dl, and encapsulated in a phospholipid bilayer membrane to form BHbV with a diameter of about 280 nm. The O(2) affinity (P(50)) of the BHbV was regulated by coencapsulation of an appropriate amount of Cl(-) (as NaCl), which binds to BHb as an allosteric effector, in the range 16-28 Torr, comparable to human blood (P(50) = 28 Torr). This is quite simple in comparison with HHb which requires phosphate derivatives such as pyridoxal 5'-phosphate as a replacement for 2,3-diphoshoglyceric acid. The viscosity and colloid osmotic pressure of the BHbV when suspended in 5% human serum albumin are 3.5 cP and 20 Torr, respectively, comparable to those of human blood. In conclusion, BHb can be used as a source for the production of HbV, not only because of its abundance in the cattle industry, but also because of the physicochemical advantages of the purification process, thermal stability, and regulation of O(2) affinity in comparison with HHb. PMID- 11927001 TI - Endostatin inhibits adhesion of endothelial cells to collagen I via alpha(2)beta(1) integrin, a possible cause of prevention of chondrosarcoma growth. AB - Endostatin derived from collagen XVIII is a potent endogenous anti-angiogenic factor that induces regression of various tumors of epithelial origin. Endostatin has been shown to inhibit endothelial cell functions, however, its effect remains controversial. We first attempted here to apply the inhibitory effect of recombinant human endostatin on chondrosarcomas, which originate from the mesenchyme, in nude mice. Endostatin induced reduction of chondrosarcoma growth and tumor angiogenesis in vivo. However, endostatin showed no effect on the proliferation and migration of chondrosarcoma cells in vitro. Next, we investigated the interactions between endostatin and endothelial cells in detail. Endostatin inhibited the migration on and attachment to collagen I but did not affect the proliferation of endothelial cells. Although the migration of endothelial cells was stimulated by angiogenic factors such as basic fibroblast growth factor and vascular endothelial growth factor, endostatin showed similar inhibitory effects on it in the presence and absence of the stimulants. Moreover, the inhibitory effect against endothelial cell attachment to collagen I was attenuated or modulated in the presence of neutralizing antibodies of alpha(2), alpha(5)beta(1), and alpha(V)beta(3) integrins but not that of alpha(1) integrin. Our results suggest that endostatin might suppress the alpha(2)beta(1) integrin function of endothelial cells via alpha(5)beta(1) or alpha(V)beta(3) integrin. We propose here that endostatin might be effective for anti-angiogenic therapy for human chondrosarcomas through the suppression of alpha(2)beta(1) integrin functions in endothelial cells. PMID- 11927002 TI - Estrone and 17beta-estradiol reverse breast cancer resistance protein-mediated multidrug resistance. AB - Breast cancer resistance protein (BCRP), an adenosine triphosphate-binding cassette transporter, confers resistance to a series of anticancer reagents, including mitoxantrone, SN-38 and topotecan. In the present study, we found that estrone and 17beta-estradiol potentiated the cytotoxicity of mitoxantrone, SN-38 and topotecan in BCRP-transduced K562 cells (K562 / BCRP). These estrogens showed only a marginal effect, or none, in parental K562 cells. Estrone and 17beta estradiol increased the cellular accumulation of topotecan in K562 / BCRP cells, but not in K562 cells, suggesting that these estrogens inhibit the BCRP-mediated drug efflux and overcome drug resistance. PMID- 11927003 TI - Food restriction inhibits the growth of intestinal polyps in multiple intestinal neoplasia mouse. AB - The effect of food restriction (FR) on spontaneous intestinal carcinogenesis in multiple intestinal neoplasia (Min) mice was examined. Thirty male Min mice were allotted to ad libitum feeding control and 20% FR groups from six weeks of age until the end of the 13-week experimental period. Although the total number of visible intestinal polyps in the FR group was not significantly different from the control group value, a significant decrease in large-sized polyps (>2 mm) and an increase in small-sized polyps (< or =2 mm) were observed in the distal small intestine. In this segment, the percentage of apoptotic cells counted in intestinal polyps in the FR group was significantly higher than in the control group, the percentage of proliferating cell nuclear antigen (PCNA)-positive cells not being significantly different. These results indicate that the FR may inhibit the growth of intestinal polyps in the Min mouse, and that apoptosis contributed in part to the inhibitory effect. PMID- 11927004 TI - Induction of apoptosis by sulindac in azoxymethane-induced possible colonic premalignant lesions in rats. AB - We have reported that beta-catenin-accumulated crypts (BCAC), which do not have the appearance of aberrant crypt foci (ACF) are possible colonic premalignant lesions in rats. Suppression of the occurrence and advancement of such lesions should have critical relevance to cancer prevention. This study examined whether sulindac, a chemopreventive nonsteroidal anti-inflammatory drug is able to induce apoptosis in such premalignant lesions. At 6 weeks of age, rats groups 1 - 3 were given azoxymethane (AOM) (15 mg/kg-body weight) once weekly for 3 weeks. Two groups were given sulindac in the diet (200 and 400 ppm), starting at 9 weeks of age. The rats were sacrificed at the termination, and the colons were carefully examined. The incidence and crypt multiplicity of BCAC and ACF were significantly less than those of the control group. The effect of sulindac on the expression of BCAC was greater than that on ACF. Exposure to sulindac significantly increased the apoptotic index (terminal deoxynucleotide transferase dUTP nick-end labeling (TUNEL)-positive cells) in BCAC. However, no significant increase of the index was found in the case of ACF. These results suggest that the chemopreventive effect of sulindac in rats is related to the induction of apoptosis in premalignant lesions. Our results also provide additional evidence that BCAC are premalignant lesions in colon carcinogenesis in rodents. PMID- 11927005 TI - Lack of direct involvement of 8-hydroxy-2'-deoxyguanosine in hypoxanthine-guanine phosphoribosyltransferase mutagenesis in V79 cells treated with N,N'-bis(2 hydroxyperoxy-2-methoxyethyl)-1,4,5,8-naphthalenetetracarboxylic-diimide (NP-III) or riboflavin. AB - The object of this study is to investigate the relationship between a typical product of oxidative DNA damage, 8-hydroxy-2'-deoxyguanosine (8OHdG), and mutagenesis in V79 cells through a molecular analysis of hypoxanthine-guanine phosphoribosyltransferase (hprt) gene mutants. We performed a direct sequencing analysis of the cDNA of mutants obtained after treatment with N,N'-bis(2 hydroxyperoxy-2-methoxyethyl)-1,4,5,8-naphthalenetetracarboxylic-diimide (NP-III) or riboflavin, each of which induces the formation of 8OHdG in cellular DNA upon UVA irradiation. The frequency of mutation after both treatments was no more than 2 to 5 times the control value. A considerable number of the mutants could not be amplified by RT-PCR, and this was also the case for the control mutants. Among the mutants analyzed, deletions and a TA-->AT transversion occurred predominantly. The reasons for the weak association of induction of 8OHdG with frequency of mutation and the possible mechanism of oxidative-stress-derived mutagenesis are discussed. PMID- 11927006 TI - Gene expression for suppressors of telomerase activity (telomeric-repeat binding factors) in breast cancer. AB - Mechanisms regulating telomerase activity and telomere length remain incompletely understood in human breast cancer. We therefore studied gene expression for telomeric-repeat binding factors (TRFs) in relation to telomerase activity, telomere length, and clinicopathologic factors in human breast cancer. Telomerase activity was detected in 65.8% of 38 breast cancers, but none of 16 noncancerous samples. Terminal restriction fragments were longer in noncancerous than in cancerous tissues, but not significantly. Among 8 patients with both cancer and paired noncancerous tissue available for terminal restriction fragments length assay, terminal restriction fragments were shorter in cancers than in paired noncancerous samples in all but one. Significantly more mRNA encoding TRF1 and 2 was detected in noncancerous than in cancer tissues. Additionally, expression of TRF1 and 2 mRNA was significantly higher in cancers without detectable telomerase activity than in cancers showing activity. Expression of these genes tended to show a negative correlation with terminal restriction fragments length, but this was not statistically significant. No correlation was seen between TRF1 or 2 mRNA expression, and clinicopathologic factors except for TRF1 with respect to tumor size and progesterone receptor status. In addition to reactivation of telomerase activity, escape from negative regulation of this activity is needed to maintain telomere length during cell proliferation in breast cancer. Genes encoding telomerase inhibitors might be of value in gene therapy against human breast cancer. PMID- 11927007 TI - Three-dimensional reconstruction and fractal geometric analysis of serrated adenoma. AB - Serrated adenoma (SA) is a relatively newly defined entity of colorectal neoplasm first characterized by Longacre and Fenoglio-Preiser in 1990. This lesion is characterized by a complicated serrated edge of crypts. In this study, we performed three-dimensional (3-D) reconstruction, including 3-D distribution patterns of Ki-67-positive cells and fractal dimension of SA, in order to evaluate the nature of the complicated architecture, including its possible morphogenesis. We studied nine colonoscopic polypectomy specimens including three SAs, three tubular adenomas (TAs), and three hyperplastic polyps (HPs). Sixty serial tissue sections per case were stained alternately with hematoxylin and eosin (H&E) and Ki-67 immunostain. Each serial image was then digitized for 3-D computer analysis and the distribution pattern of Ki-67-positive cells was evaluated. Ki-67-immunostained sections were also subjected to 2-D quantitative morphometric study. In addition, the fractal dimensions of images from H&E stained sections were examined using a box-counting method. Results of the 3-D reconstruction study demonstrated that glandular budding and branching were more frequent in SA than in TA or HP. These findings were confirmed quantitatively by the results of fractal geometric analysis of these polyps (fractal dimension:1.34 +/- 0.08 for SA, 1.23 +/- 0.07 for TA, and 1.28 +/- 0.12 for HP). Ki-67-positive cells in HP were localized mainly in the bottom of crypts and those in TA were diffusely distributed, while Ki-67-positive cells in SA were mainly aggregated in the depressed sites of serrated epithelia. These findings were also confirmed quantitatively using 2-D morphometry. These distribution patterns of the proliferative zone of SA are considered to contribute to the formation of the characteristic serrated epithelia and the complicated morphological appearance of SA. PMID- 11927008 TI - Mutations of p53, c-kit, K-ras, and beta-catenin gene in non-Hodgkin's lymphoma of adrenal gland. AB - Malignant lymphoma of the adrenal gland is a rare disease, usually with diffuse large cell morphology and B-cell immunophenotype, and often associated with Epstein-Barr virus infection. In this study, mutations of p53, c-kit, K-ras, and beta-catenin gene were analyzed in 17 cases (13 males and four females with ages ranging from 25 to 84 years) of such lymphomas by polymerase chain reaction single strand conformation polymorphism followed by direct sequencing. Selected exons in each gene, representing hot spots, were analyzed. All 44 mutations detected were single-nucleotide substitutions and 33 were missense mutations. Nineteen mutations were detected in exon 5 and / or 7 of the p53 gene in nine of 17 cases (52.9%) and 21 in exon 11 and / or 17 of the c-kit gene in 10 of 14 cases (71.4%). Bilateral adrenal lesions in one case who had not received any adjuvant therapy showed different mutational patterns of the p53 and c-kit genes, suggesting different clonal evolution of lymphoma between the left and right sides. Mutation at codon 13 of the K-ras gene was detected in one of 14 cases (7.1%), and in exon 3 of the beta-catenin gene in three of 12 cases (25%). All but one mutation were transition mutations, indicating that some endogenous mutagens act in lymphomagenesis in the adrenal gland. Our results suggest that p53 and c-kit gene mutations might play a role in adrenal lymphomagenesis. PMID- 11927009 TI - Suppression of postmitochondrial signaling and delayed response to UV-induced nuclear apoptosis in HeLa cells. AB - Activation of postmitochondrial pathways by UV irradiation was examined using mouse lymphoma 3SB and human leukemic Jurkat cells and two human carcinoma cell lines (HeLa and MCF-7). Exposure of 3SB and Jurkat cells resulted in large amounts of cytochrome c and apoptosis-inducing factor (AIF) being released into the cytosol, and a clear laddering pattern of DNA fragments was observed within 3 h of incubation after irradiation. Simultaneously, activation of caspase-9 and its downstream caspases was detected. HeLa and MCF-7 cells also showed extensive release of mitochondrial factors and caspase-9 activation at 4 to 6 h after exposure, but apoptotic nuclear changes appeared much later. Compared with 3SB and Jurkat cells, these carcinoma cell lines exhibited reduced activation of caspase-9-like proteolytic activity by UV radiation, and levels of caspase-3-like activity in HeLa cells were extremely low, similar to those in caspase-3 deficient MCF-7 cells. These results suggest that the delayed response to UV induced nuclear apoptosis in HeLa cells is due to a reduced activation of the caspase cascade downstream of cytochrome c release and suppression of caspase-3 activity. PMID- 11927010 TI - Multiplexed analysis of post-PCR fluorescence-labeled microsatellite alleles and statistical evaluation of their imbalance in brain tumors. AB - Detection of the loss of chromosomal regions in cancerous tissues has diagnostic and prognostic relevance, and the development of a reliable and cost-effective technique for this is clinically important. Here we present an efficient technique for quantitative detection of microsatellite alleles, using a post-PCR fluorescence-labeling procedure and multiplexed analysis. We also present a new statistical method for the interpretation of the data that permits reliable and sensitive evaluation of the allelic status of sampled DNA. A high-resolution analysis of allelic imbalance on chromosomes 1p, 10 and 19q in 28 glioma samples of various types using this method revealed that allelic imbalances are more frequent than have been reported, suggesting the diagnostic value of this method in examining the genetic profiles of gliomas. PMID- 11927011 TI - Prognostic significance of matrix metalloproteinase-7 (MMP-7) expression at the invasive front in gastric carcinoma. AB - To evaluate the clinicopathological significance of matrix metalloproteinase-7 (MMP-7) expression in gastric carcinoma, we investigated immunohistochemically MMP-7 expression in 214 gastric carcinomas, and examined its relations with the clinicopathologic parameters including patient prognosis. MMP-7 expressed predominantly in cancer cells, and MMP-7-positive tumor cells were preferentially found in deeply invading nests, especially at the invasive front. The mean MMP-7 labeling index (LI) at the invasive front was significantly higher in tumors invading or penetrating the muscularis propria and in stages II - IV than within the submucosal layer and in stage I, respectively (P < 0.001). Statistical analysis revealed that MMP-7 LI at the invasive front was related to lymph node metastasis, vascular invasion, and lymphatic permeation, when all 214 cases were examined as one group (P < 0.05 for all), and the cases with high MMP-7 expression at the invasive front showed significantly more unfavorable prognosis as compared with that of low MMP-7 expression tumors (P < 0.01). Multivariate analysis revealed that TNM stage and MMP-7 expression status at the invasive front were independent prognostic factors (P = 0.0017, relative risk (RR) = 3.12; P = 0.0019, RR = 2.67, respectively). Our findings indicated that expression of MMP-7 at the invasive front is closely associated with local invasiveness, and might be a reliable prognostic marker for patients with gastric carcinoma. PMID- 11927012 TI - VCP (p97) regulates NFkappaB signaling pathway, which is important for metastasis of osteosarcoma cell line. AB - In order to identify genes associated with metastasis, suppression subtractive hybridization (SSH) was performed using murine osteosarcoma cell line Dunn and its subline with higher metastatic potential, LM8. SSH revealed expression of the gene encoding valosin-containing protein (VCP; also known as p97) to be constitutively activated in LM8 cells, but it declined in Dunn cells when the cells became confluent. Because VCP is known to be involved in the ubiquitination process of Inhibitor-kappaBalpha (IkappaBalpha), an inhibitor of nuclear factor kappaB (NFkappaB), whether VCP influences NFkappaB activation or not was examined by using VCP-transfected Dunn cells (Dunn/VCPs). When stimulated with tumor necrosis factor-alpha (TNFalpha), Dunn/VCPs showed constantly activated NFkappaB, although in the original Dunn cells and control vector transfectant (Dunn/Dunn-c) NFkappaB activation ceased when the cells became confluent. Western immunoblot analysis showed an increase of phosphorylated IkappaBalpha (p-IkappaBalpha) in the cytoplasm of confluent Dunn/Dunn-c cells compared to that of Dunn/VCPs. Therefore, decrease of p-IkappaBalpha degrading activity might be responsible for the decrease in NFkappaB activation. In vitro apoptosis assay demonstrated increased apoptosis rates of Dunn/Dunn-c cells after TNFalpha stimulation compared to those of Dunn/VCPs and LM8 cells. In vivo metastasis assay showed increased incidences of metastatic events in Dunn/VCP-1 inoculated male C3H mice compared to those in Dunn/Dunn-c inoculated mice. These findings suggested that VCP expression plays an important role in the metastatic process. Anti-apoptotic potential in these cells owing to constant NFkappaB activation via efficient cytoplasmic p-IkappaBalpha degrading activity may explain the increased metastatic potential of these cells. PMID- 11927013 TI - Evaluation of an indicator for lymph node metastasis of esophageal squamous cell carcinoma invading the submucosal layer. AB - Lymph node metastasis is a major prognostic factor for esophageal squamous cell carcinoma (ESCC). In recent years, endoscopic mucosal resection (EMR) has been developed with excellent results for the treatment of the superficial ESCC. To make the EMR treatment successful, it is important to establish a good indicator to identify ESCC patients at a high risk of lymph node metastasis. In this study, we examined clinicopathological and immunohistochemical factors to investigate the factors involved in lymph node metastasis of ESCC invading to the submucosal layer (sm-ESCC). Surgical specimens from 84 sm-ESCC patients were examined. Among 84 sm-ESCC patients, 33 (39.3%) had lymph node metastases. Clinicopathologically, tumor depth, lymphatic invasion and blood vessel invasion showed significant correlations with lymph node metastasis by univariate analysis. Tumor depth and lymphatic invasion showed significant correlations by multivariate analysis of these factors. Immunohistochemically, P53 accumulation was observed in 45 cases (53.6%), cyclin D1 overexpression in 25 (29.8%), and pRB in 65 (77.4%). P53 accumulation, cyclin D1 overexpression and MIB-1 Labeling Index were significantly associated with lymph node metastasis by univariate analysis, and P53 accumulation showed a significant correlation with lymph node metastasis by multivariate analysis. Among tumor depth, lymphatic invasion and P53 accumulation, tumor depth and lymphatic invasion were significantly correlated with lymph node metastasis (P = 0.0023 and P = 0.0092, respectively) by multivariate analysis. These data suggest that tumor depth and lymphatic invasion can be considered as good indicators for lymph node metastasis among patients with sm-ESCC. In addition, P53 accumulation could be helpful to identify the patients who need additional treatment after EMR. PMID- 11927014 TI - Selective expansion of human natural killer cells from peripheral blood mononuclear cells by the cell line, HFWT. AB - An anchorage-dependent Wilms tumor cell line HFWT was found to stimulate selective and remarkable expansion of human natural killer (NK) cells from human peripheral blood mononuclear cells (PBMC). After PBMC of healthy donors were cultured on irradiated HFWT cells for 10 - 21 days, the lymphocytes expanded 58- to 401-fold. This NK cell expansion required direct contact of PBMC with live, but not fixed, HFWT cells. The PBMC from an end-stage brain tumor patient also expanded 156-fold, whereas those cultured with irradiated NK-sensitive K562 grew only 30.5-fold. CD16+ CD56+ NK cells accounted for more than 70% of the population expanded on HFWT cells. No essential difference in expression of NK receptors was observed in the expanded NK cells on HFWT and K562 and without feeder cells. The expanded NK cells killed not only fresh HFWT cells but, unexpectedly, also MHC class I-expressing autologous brain tumor cells at an effector/target ratio of 4 for 24 h. These results will contribute to the development of a large-scale preparation method for human NK cells, which will aid studies of NK cell biology and possible treatment of brain tumors. PMID- 11927015 TI - Close association between Fas ligand (FasL; CD95L)-positive tumor-associated macrophages and apoptotic cancer cells along invasive margin of colorectal carcinoma: a proposal on tumor-host interactions. AB - Anti-tumor immune responses are considered to be one of the key host reactions in human colorectal cancer, with T cells as important effector cells. For the induction of tumor-specific immunity, processing of cancer cells and priming of T cells by antigen-presenting cells are important. The present study was designed to clarify the relationship between Fas ligand (FasL; CD95L) expression and apoptotic cancer cells. Immunohistochemistry using frozen sections taken from 58 patients with colorectal cancer revealed that stromal cells composed mainly of tumor-associated macrophages expressed FasL, leaving cancer cells negative for FasL. These macrophages were abundantly distributed along the invasive margin. In situ hybridization revealed that these macrophages as well as cancer cells expressed FasL mRNA, whereas macrophages in the normal colon mucosa rarely expressed FasL. Apoptotic cancer cells recognized by monoclonal antibody M30 CytoDEATH were localized not only in cancer cell nests, but also in the stroma along the invasive margin showing a dissociated pattern, which was particularly evident in the areas of FasL+ macrophages. Furthermore, these macrophages, phenotypically similar to dendritic cells, occasionally contained M30+ apoptotic cancer cells in the cytoplasm. Clinicopathologic analyses in 123 cases revealed 1) a positive correlation between the degree of dissociated M30+ apoptotic cancer cells and the number of macrophages along the invasive margin and 2) an inverse association between the degree of dissociated M30+ apoptotic cancer cells and the occurrence of hematogenous metastasis after surgical resection of the primary tumor. In conclusion, the present study shows the importance of FasL+ activated macrophages as one of the host defense mechanisms against cancer cell spread in human colorectal cancer. PMID- 11927016 TI - The roles of JNK1 and Stat3 in the response of head and neck cancer cell lines to combined treatment with all-trans-retinoic acid and 5-fluorouracil. AB - We have used a combination of vitamin A (all-trans-retinyl palmitate), 5 fluorouracil (5-FU) and radiation to treat human head and neck squamous cell carcinoma (HNSCC). This chemoradiotherapy is called "FAR therapy." In this study we examined the effects of all-trans-retinoic acid (ATRA), the active metabolite of vitamin A, and ATRA plus 5-FU on two HNSCC cell lines (YCU-N861 and YCU-H891) to gain insight into the molecular mechanisms of FAR therapy. ATRA at 1 mM (the order of concentration found in HNSCC tumors treated with FAR therapy) inhibited cell proliferation and caused G1 cell cycle arrest in both cell lines. This was associated with a decrease in cyclin D1, an increase in p27(Kip1) and a reduction in the hyperphosphorylated form of retinoblastoma protein (pRB). With YCU-N861 cells, ATRA also caused a decrease in Bcl-2 and Bcl-X(L) and an increase in Bax. Both ATRA and 5-FU activated c-Jun N-terminal kinase (JNK) 1 and the combination of both agents resulted in additive or synergistic activation of JNK1, and also enhanced the induction of apoptosis. The YCU-H891 cells, in which the epidermal growth factor receptor (EGFR)-signal transducer and activator of transcription 3 (Stat3) pathway is constitutively activated, were more resistant to treatments with ATRA, 5-FU and the combination of both agents than YCU-N861 cells. A dominant negative Stat3 construct strongly enhanced the cellular sensitivity of this cell line to 5-FU but not to ATRA. In addition there is evidence that activation of Stat3 is associated with cellular resistance to radiation in HNSCC. Therefore, the addition to FAR therapy of agents that inhibit activation of the Stat3 pathway may enhance the clinical response of patients with HNSCC to FAR therapy. PMID- 11927017 TI - Expression of thymidine phosphorylase in primary human renal cell carcinoma by ELISA method. AB - Thymidine phosphorylase (TP) expression in 100 paired samples of renal cell carcinoma (RCC) and normal adjacent tissue was analyzed by an ELISA method. We also investigated whether TP expression correlates with clinicopathological findings and clinical outcomes of these patients. Median TP expression was 9-fold (range, 0.5-56) higher in primary tumor than in non-cancerous renal tissue (P < 0.0001). There was a significant difference with respect to tumor venous invasion. TP expression was significantly higher in patients with such venous invasion than in those without (P = 0.018). However, there was no correlation between TP level and other clinicopathological findings and the survival curves. These results suggest that ELISA is useful for evaluating TP expression of human RCC and may provide a novel approach to therapy for patients with RCC. PMID- 11927018 TI - Feasibility of national surveillance of health-care-associated infections in home care settings. AB - This article examines the rationale and strategies for surveillance of health care-associated infections in home-care settings, the challenges of nonhospital based surveillance, and the feasibility of developing a national surveillance system. PMID- 11927019 TI - Human campylobacteriosis in developing countries. AB - Campylobacteriosis is a collective description for infectious diseases caused by members of the bacterial genus Campylobacter. The only form of campylobacteriosis of major public health importance is Campylobacter enteritis due to C. jejuni and C. coli. Research and control efforts on the disease have been conducted more often in developed countries than developing countries. However, because of the increasing incidence, expanding spectrum of infections, potential of HIV-related deaths due to Campylobacter, and the availability of the complete genome sequence of C. jejuni NCTC 11168, interest in campylobacteriosis research and control in developing countries is growing. We present the distinguishing epidemiologic and clinical features of Campylobacter enteritis in developing countries relative to developed countries. National surveillance programs and international collaborations are needed to address the substantial gaps in the knowledge about the epidemiology of campylobacteriosis in developing countries. PMID- 11927020 TI - Immunization with heterologous flaviviruses protective against fatal West Nile encephalitis. AB - Prior immunization of hamsters with three heterologous flaviviruses (Japanese encephalitis virus [JEV] SA14-2-8 vaccine, wild-type St. Louis encephalitis virus [SLEV], and Yellow fever virus [YFV] 17D vaccine) reduces the severity of subsequent West Nile virus (WNV) infection. Groups of adult hamsters were immunized with each of the heterologous flaviviruses; approximately 30 days later, the animals were injected intraperitoneally with a virulent New York strain of WNV. Subsequent levels of viremia, antibody response, and deaths were compared with those in nonimmune (control) hamsters. Immunity to JEV and SLEV was protective against clinical encephalitis and death after challenge with WNV. The antibody response in the sequentially infected hamsters also illustrates the difficulty in making a serologic diagnosis of WNV infection in animals (or humans) with preexisting Flavivirus immunity. PMID- 11927021 TI - Associations between indicators of livestock farming intensity and incidence of human Shiga toxin-producing Escherichia coli infection. AB - The impact of livestock farming on the incidence of human Shiga toxin-producing Escherichia coli (STEC) infection was assessed by using several livestock density indicators (LDI) that were generated in a systematic approach. A total of 80 LDI were considered suitable proxy measures for livestock density. Multivariate Poisson regression identified several LDI as having a significant spatial association with the incidence of human STEC infection. The strongest associations with human STEC infection were the ratio of beef cattle number to human population and the application of manure to the surface of agricultural land by a solid spreader and by a liquid spreader. This study demonstrates the value of using a systematic approach in identifying LDI and other spatial predictors of disease. PMID- 11927022 TI - Serologic evidence of Lyssavirus infections among bats, the Philippines. AB - Active surveillance for lyssaviruses was conducted among populations of bats in the Philippines. The presence of past or current Lyssavirus infection was determined by use of direct fluorescent antibody assays on bat brains and virus neutralization assays on bat sera. Although no bats were found to have active infection with a Lyssavirus, 22 had evidence of neutralizing antibody against the Australian bat lyssavirus (ABLV). Seropositivity was statistically associated with one species of bat, Miniopterus schreibersi. Results from the virus neutralization assays are consistent with the presence in the Philippines of a naturally occurring Lyssavirus related to ABLV. PMID- 11927023 TI - Novel cryptosporidium genotypes in sporadic cryptosporidiosis cases: first report of human infections with a cervine genotype. AB - In this study, we genotyped parasites from the fecal specimens of sporadic cryptosporidiosis cases in British Columbia from 1995 to 1999. Genotyping was conducted by polymerase chain amplification of the internal transcribed spacer region, a hypervariable region in the 18S rRNA gene and the Cryptosporidium oocyst wall protein gene. Subsequent analysis was by restriction fragment length polymorphism and DNA sequencing. We identified two new Cryptosporidium genotypes in humans. One of these genotypes has been found recently in deer in New York state. The other genotype has not been identified in humans or animals. These results have important implications for drinking water quality strategies, especially for communities that obtain drinking water supplies from surface sources located in forested regions with deer populations. PMID- 11927024 TI - Molecular epidemiology of adenovirus type 7 in the United States, 1966-2000. AB - Genetic variation among 166 isolates of human adenovirus 7 (Ad7) obtained from 1966 to 2000 from the United States and Eastern Ontario, Canada, was determined by genome restriction analysis. Most (65%) isolates were identified as Ad7b. Two genome types previously undocumented in North America were also identified: Ad7d2 (28%), which first appeared in 1993 and was later identified throughout the Midwest and Northeast of the United States and in Canada; and Ad7h (2%), which was identified only in the U.S. Southwest in 1998 and 2000. Since 1996, Ad7d2 has been responsible for several civilian outbreaks of Ad7 disease and was the primary cause of a large outbreak of respiratory illness at a military recruit training center. The appearance of Ad7d2 and Ad7h in North America represents recent introduction of these viruses from previously geographically restricted areas and may herald a shift in predominant genome type circulating in the United States. PMID- 11927025 TI - A European study on the relationship between antimicrobial use and antimicrobial resistance. AB - In Europe, antimicrobial resistance has been monitored since 1998 by the European Antimicrobial Resistance Surveillance System (EARSS). We examined the relationship between penicillin nonsusceptibility of invasive isolates of Streptococcus pneumoniae and antibiotic sales. Information was collected on 1998 99 resistance data for invasive isolates of S. pneumoniae to penicillin, based on surveillance data from EARSS and on outpatient sales during 1997 for beta-lactam antibiotics and macrolides. Our results show that in Europe antimicrobial resistance of S. pneumoniae to penicillin is correlated with use of beta-lactam antibiotics and macrolides. PMID- 11927026 TI - Eastern equine encephalomyelitis virus infection in a horse from California. AB - A yearling quarter horse, which was raised in southern California, received routine vaccinations for prevention of infection by Eastern equine encephalomyelitis virus (EEEV). One week later, severe neurologic signs developed, and the horse was humanely destroyed. A vaccine-related encephalomyelitis was later suspected. A final diagnosis of EEEV infection was established on the basis of acute onset of the neurologic signs, histopathologic and serologic testing, and isolation and molecular characterization of EEEV from brain tissue. The vaccine was extensively tested for viral inactivation. Nucleotide sequences from the vaccine and the virus isolated in the affected horse were also compared. In California, arboviral encephalomyelitides are rarely reported, and EEEV infection has not previously been documented. This report describes the occurrence of EEEV infection in the horse and the investigation to determine the source of infection, which was not definitively identified. PMID- 11927027 TI - Predicting the risk of Lyme disease: habitat suitability for Ixodes scapularis in the north central United States. AB - The distribution and abundance of Ixodes scapularis were studied in Wisconsin, northern Illinois, and portions of the Upper Peninsula of Michigan by inspecting small mammals for ticks and by collecting questing ticks at 138 locations in state parks and natural areas. Environmental data were gathered at a local level (i.e., micro and meso levels), and a geographic information system (GIS) was used with several digitized coverages of environmental data to create a habitat profile for each site and a grid map for Wisconsin and Illinois. Results showed that the presence and abundance of I. scapularis varied, even when the host population was adequate. Tick presence was positively associated with deciduous, dry to mesic forests and alfisol-type soils of sandy or loam-sand textures overlying sedimentary rock. Tick absence was associated with grasslands, conifer forests, wet to wet/mesic forests, acidic soils of low fertility and a clay soil texture, and Precambrian bedrock. We performed a discriminant analysis to determine environmental differences between positive and negative tick sites and derived a regression equation to examine the probability of I. scapularis presence per grid. Both analyses indicated that soil order and land cover were the dominant contributors to tick presence. We then constructed a risk map indicating suitable habitats within areas where I. scapularis is already established. The risk map also shows areas of high probability the tick will become established if introduced. Thus, this risk analysis has both explanatory power and predictive capability. PMID- 11927028 TI - Molecular classification of enteroviruses not identified by neutralization tests. AB - We isolated six viruses from patients diagnosed with aseptic meningitis or hand, foot, and mouth disease. The cytopathic effect of these viruses on cultured cells was like that of enteroviruses. However, viral neutralization tests against standard antisera were negative. Phylogenetic analysis with the complete VP4 nucleotide sequences of these 6 viruses and 29 serotypes of enteroviruses classified 3 of the viruses as serotype echovirus type 18 (EV18) and 3 as serotype human enterovirus 71 (HEV71). These results were confirmed by remicroneutralization tests with HEV-monospecific antisera or an additional phylogenetic analysis with the complete VP4 nucleotide sequences. Phylogenetic analysis with complete VP4 genes is more useful than neutralization tests with enterovirus serotype-specific antisera in identifying enterovirus serotypes. PMID- 11927030 TI - Epidemiologic features of four successive annual outbreaks of bubonic plague in Mahajanga, Madagascar. AB - From 1995 to 1998, outbreaks of bubonic plague occurred annually in the coastal city of Mahajanga, Madagascar. A total of 1,702 clinically suspected cases of bubonic plague were reported, including 515 laboratory confirmed by Yersinia pestis isolation (297), enzyme-linked immunosorbent assay, or both. Incidence was higher in males and young persons. Most buboes were inguinal, but children had a higher frequency of cervical or axillary buboes. Among laboratory-confirmed hospitalized patients, the case-fatality rate was 7.9%, although all Y. pestis isolates were sensitive to streptomycin, the recommended antibiotic. In this tropical city, plague outbreaks occur during the dry and cool season. Most cases are concentrated in the same crowded and unsanitary districts, a result of close contact among humans, rats, and shrews. Plague remains an important public health problem in Madagascar, and the potential is substantial for spread to other coastal cities and abroad. PMID- 11927031 TI - Rickettsia felis in Ctenocephalides spp. fleas, Brazil. AB - In June 2000, suspected cases of Brazilian spotted fever (BSF) occurred in Coronel Fabriciano Municipality, Minas Gerais State, Brazil. Pooled fleas collected near two fatal cases contained rickettsial DNA. The nucleotide sequence alignment of the 391-bp segment of the 17-kDa protein gene showed that the products were identical to each other and to the R. felis 17-kDa gene, confirming circulation of R. felis in Brazil. PMID- 11927029 TI - Listeria monocytogenes infection in Israel and review of cases worldwide. AB - Listeria monocytogenes, an uncommon foodborne pathogen, is increasingly recognized as a cause of life-threatening disease. A marked increase in reported cases of listeriosis during 1998 motivated a retrospective nationwide survey of the infection in Israel. From 1995 to 1999, 161 cases were identified; 70 (43%) were perinatal infections, with a fetal mortality rate of 45%. Most (74%) of the 91 nonperinatal infections involved immunocompromised patients with malignancies, chronic liver disease, chronic renal failure, or diabetes mellitus. The common clinical syndromes in these patients were primary bacteremia (47%) and meningitis (28%). The crude case-fatality rate in this group was 38%, with a higher death rate in immunocompromised patients. PMID- 11927033 TI - Enzootic Angiostrongylus cantonensis in rats and snails after an outbreak of human eosinophilic meningitis, Jamaica. AB - After an outbreak in 2000 of eosinophilic meningitis in tourists to Jamaica, we looked for Angiostrongylus cantonensis in rats and snails on the island. Overall, 22% (24/109) of rats harbored adult worms, and 8% (4/48) of snails harbored A. cantonensis larvae. This report is the first of enzootic A. cantonensis infection in Jamaica, providing evidence that this parasite is likely to cause human cases of eosinophilic meningitis. PMID- 11927032 TI - Severe Ehrlichia chaffeensis infection in a lung transplant recipient: a review of ehrlichiosis in the immunocompromised patient. AB - We describe a case of human ehrlichiosis in a lung transplant recipient and review published reports on ehrlichiosis in immunocompromised patients. Despite early therapy with doxycycline, our patient had unusually severe illness with features of thrombotic thrombocytopenic purpura. Of 23 reported cases of ehrlichiosis in immunocompromised patients, organ failure occurred in all patients and 6 (25%) died. PMID- 11927034 TI - Recent increase in meningitis Caused by Neisseria meningitidis serogroups A and W135, Yaounde, Cameroon. AB - From 1991 to 1998, Neisseria meningitidis serogroups A, B, and C represented 2% 10% of strains isolated from cases of bacterial meningitis in Yaounde. During 1999 to 2000, the percentage of meningococci reached 17%, a proportion never reported since recordkeeping began in 1984. The increase of serogroup A meningococci and the emergence of W135 strains highlight the need for increased surveillance for better diagnosis and prevention. PMID- 11927035 TI - HIV prevalence in a gold mining camp in the Amazon region, Guyana. AB - The prevalence of HIV infection among men in a gold mining camp in the Amazon region of Guyana was 6.5%. This high percentage of HIV infection provides a reservoir for the virus in this region, warranting immediate public health intervention to curb its spread. As malaria is endemic in the Amazon Basin (>30,000 cases/year), the impact of coinfection may be substantial. PMID- 11927036 TI - Hajj-related Neisseria meningitidis serogroup w135 in Mauritius. AB - Meningococcal disease is rare in Mauritius; only one case was reported from 1992 to 1999. However, since June 2000, four cases have occurred. Epidemiologic information and typing results indicate that these recent cases probably followed the introduction of Neisseria meningitidis W135 in Mauritius by pilgrims returning from the Hajj in 2000 and 2001. PMID- 11927037 TI - Contagion on the Internet. PMID- 11927038 TI - Emergence and rapid spread of tetracycline-resistant Vibrio cholerae strains, Madagascar. PMID- 11927039 TI - Clinical Trials Report. PMID- 11927040 TI - Update on the Management of Hepatitis C in Liver Transplant Recipients. AB - Hepatic failure due to hepatitis C is the leading indicator for orthotopic liver transplantation (OLT) in the United States. Unfortunately, recurrent hepatitis C virus infection is essentially universal following orthotopic liver transplantation. Although significant advances have been made in the past decade for the treatment of hepatitis C, a similar level of success has not yet been achieved for most hepatitis C virus-infected liver transplant recipients. In addition, deleterious side effects of the currently available antiviral agents continue to significantly hamper their use. Several recent reports, however, indicate that newer immunosuppressive regimens combined with novel modifications of existing treatment paradigms will likely lead to improved clinical outcomes for the hepatitis C virus-infected liver transplant recipient. PMID- 11927041 TI - Mechanisms of Emerging Diarrheagenic Escherichia coli Infection. AB - Diarrheagenic Escherichia coli organisms are major causes of morbidity and mortality worldwide. Although most strains of E. coli are harmless commensals, a few types have emerged that are capable of disrupting the normal physiology of the human gut, producing illness ranging from watery diarrhea to fatal hemorrhagic colitis. Diarrheagenic E. coli cause infection by a variety of complex mechanisms, some of which are incompletely understood. These include adherence, elaboration of toxigenic mediators, invasion of the intestinal mucosa, and transportation of bacterial proteins into the host cells. Specific components of the host-microbial interaction that cause damage have been identified, increasing our understanding of the mechanisms of diarrhea. This article reviews some of the recent findings about the pathogenesis and infectious processes involved in three emerging pathotypes of this fascinating gram-negative bacterium. PMID- 11927042 TI - Overview of Hepatitis E Virus. AB - Hepatitis E virus (HEV) is an enterically transmitted virus usually presenting as an acute self-limiting disease. However, mortality increases dramatically from around 1% to 20% in pregnant women. HEV has been the cause of very large outbreaks of hepatitis in developing countries and is also responsible for a significant number of sporadic cases. It is clear that cases occur outside the endemic areas, and new isolates have been identified. HEV-like viruses have also been found in various animal groups, and it is likely that HEV can be regarded as a zoonotic infection. Preventative measures at the moment depend mainly on the provision of clean water supplies, although a vaccine is now undergoing clinical trials. PMID- 11927043 TI - Entamoeba histolytica: Clinical Update and Vaccine Prospects. AB - Amebiasis is extraordinarily common in children of the developing world. This realization has come from application of diagnostic techniques that distinguish the nonpathogenic parasite Entamoeba dispar from Entamoeba histolytica. E. histolytica infection is found in children with dysentery, diarrhea, and in many cases in children with no gastrointestinal symptoms. Genetically distinct strains of E. histolytica exist but evidence is too preliminary to judge if some strains are more virulent than others. A provocative study from Tanzania has shown that pregnant women with HIV infection who are coinfected with E. histolytica are at greater risk of delivering a low birth weight infant. Perhaps the most exciting new data is the identification of acquired immunity to recurrent E. histolytica infection. Acquired immunity is linked to a mucosal immune response against a major virulence factor of the parasite, a Gal/GalNAc lectin responsible for adherence and killing of the host. Prospects for a vaccine are thus brightening at the same time that the true burden of disease is comprehended. PMID- 11927044 TI - Clinical Trials Report. PMID- 11927045 TI - The Development of Microbicides for Clinical Use to Prevent Sexually Transmitted Diseases. AB - Approximately 60 vaginal microbicides are under development for the prevention of HIV/AIDS and other sexually transmitted pathogens. The history and current status of the field are discussed with emphasis on the lessons learned from recent clinical trials, along with an emphasis on the mechanisms involved in the sexual transmission of HIV and how this information influences microbicide development. Additionally, the current status of in vitro and animal systems used for evaluating microbicide efficacy, as well as the challenges involved in developing more appropriate and practical assays, are discussed. Also discussed are the challenges that face the microbicide product development field in meeting US Food and Drug Administration requirements regarding product safety and stability. PMID- 11927046 TI - Bacterial Vaginosis Complicating Pregnancy and Gynecologic Surgery. AB - Bacterial vaginosis, the most commonly diagnosed vaginitis, has traditionally been regarded as a benign condition. However, recent evidence suggests association with preclinical miscarriage, early pregnancy loss, preterm labor, low birth weight infants, gynecologic surgical infections, and postabortal pelvic inflammatory diseases. Since studies show significant reduction in preterm labor and postabortal endometritis with treatment, it is recommended that bacterial vaginosis in high-risk women for preterm labor and women who undergo surgical abortion be treated regardless of symptoms. No treatment is recommended, however, for asymptomatic low-risk women. PMID- 11927047 TI - Update on Quinolone Resistance in Neisseria gonorrhoeae. AB - Quinolones are widely used for treating gonococcal infections, typically in single-dose, oral regimens. However, in the 1990s, quinolone-resistant Neisseria gonorrhoeae emerged, potentially compromising the utility of this drug class. In the past year, these strains have widely disseminated, accounting for over half of isolates in parts of Southeast Asia. The molecular mechanism of resistance has been localized to multiple mutations in genes coding for the bacterial DNA gyrase and topoisomerase enzymes. These mutations accumulate until the minimum inhibitory concentration is 4.0 g/mL or more, which in clinical studies appears to be the threshold for clinical treatment failure. Quinolone-resistant N. gonorrhoeae is independent from other plasmid- and chromosomally-mediated resistance determinants; nearly all isolates to date have been sensitive to cephalosporins and spectinomycin. Nevertheless, designing public health strategies to contain quinolone-resistant N. gonorrhoeae will be difficult. PMID- 11927048 TI - The Use of Urine and Self-obtained Vaginal Swabs for the Diagnosis of Sexually Transmitted Diseases. AB - Studies have reported that self-collected specimens, such as urine or vaginal swabs, can be successfully used to diagnose sexually transmitted infections when they are used with nucleic acid amplification assays. This eliminates the necessity for a clinician-performed pelvic examination for women, or a urethral swab for men, for sample collection. These nucleic acid amplification assays used with self-collected specimens are highly sensitive and specific, and their use may be extended to broad nonclinic screening venues, where their use can augment public health programs designed to control the epidemic of sexually transmitted diseases. PMID- 11927050 TI - Structured Treatment Interruption: Approaches and Risks. AB - Although highly active antiretroviral therapy suppresses HIV replication resulting in extraordinary clinical benefits, toxicity, adherence difficulties, and the monetary cost of medications limit the long-term effectiveness and availability of therapy for many HIV-infected individuals. Strategies to interrupt therapy have been proposed as a means to enhance the sustainability of antiretroviral treatment. Widely different approaches with varied patient populations, theoretical concepts, and clinical designs are frequently lumped together as "structured treatment interruptions." This review summarizes the approaches and risks of treatment interruptions in HIV infection. Currently, none of these strategies can be recommended in standard clinical practice. PMID- 11927049 TI - Management of Opportunistic Infection Prophylaxis in the Highly Active Antiretroviral Therapy Era. AB - Prophylaxis and maintenance therapy against opportunistic infections are a mainstay of management of HIV-infected patients and have led to a significant improvement in quality of life and survival. Antiretroviral combination therapy (ART) has markedly changed the natural course of HIV infection. Incidence of opportunistic infections (OIs) has declined and survival after an OI has improved. Achieving a CD4 count of 200 cells/L after 6 months of ART is a valuable marker for low risk of OI afterwards. Therefore, recommendations on prophylaxis and maintenance therapy need to be redefined. Criteria for discontinuation, such as a CD4 count rise above threshold values and time above threshold values as response to ART, should be evaluated for the most frequent OIs. Reliable data in favor of discontinuation of primary prophylaxis against Pneumocystis carinii pneumonia, toxoplasmic encephalitis, and Mycobacterium avium infection have been published. Discontinuation of maintenance therapy against P. carinii pneumonia is possible, and may be safe against cytomegalovirus retinitis, M. avium, and cryptococcosis and toxoplasmosis in selected patients. Pharmacologic interactions between drugs used for OI prophylaxis and antiretroviral drugs need to be taken into account. PMID- 11927052 TI - How accurate is the Canadian C-spine rule for the detection of clinically significant cervical spine injuries? PMID- 11927051 TI - Are new antihypertensive agents better than old antihypertensive agents in preventing cardiovascular complications? PMID- 11927053 TI - How effective are weekly antenatal steroids for decreasing the risks associated with preterm delivery? PMID- 11927054 TI - What is the most effective regimen for eradication of Helicobacter pylori in patients who have failed a first eradication attempt? PMID- 11927055 TI - Clinical inquiries. Does ambulatory blood pressure monitoring aid in the management of patients with hypertension? PMID- 11927056 TI - Clinical inquiries. What is the most effective treatment for tinea pedis (athlete's foot)? PMID- 11927057 TI - Clinical inquiries. What is the initial approach to the treatment of shoulder pain? PMID- 11927058 TI - Minor acute illness: a preliminary research report on the "worried well". AB - OBJECTIVES: Our objectives were to determine how patients who make frequent use of the medical system (high users) with medically unexplained symptoms met our chart-rating criteria for somatization and minor acute illness and what the stability of such diagnoses were over time. STUDY DESIGN: A chart review was performed at baseline and 1 and 2 years; we re-rated the charts of patients initially rated as having somatization, as well as a 15% sample of those with minor acute illness. POPULATION: We obtained a random sample of high-use patients (= 6 visits/year) aged 21 to 55 years who were identified from the management information system. OUTCOMES: We measured chart review designations as organic disease, somatization, or minor acute illness. RESULTS: Among 883 high users at baseline, 35% had organic diseases; 14% had somatization; and 51% had minor acute illness as their primary problems. No patients with initial minor acute diagnoses were reclassified as having somatization 1 or 2 years later, and all but 2 patients had minor acute illness in 1 or both follow-up years. CONCLUSIONS: Minor acute illness was more common among high users than somatization and organic diseases combined. It has not previously been studied but probably has been recognized by clinicians as the "worried well." Diagnoses of somatization were unstable over 2 years follow-up, while minor acute diagnoses were stable, supporting the latter as a valid entity. PMID- 11927059 TI - Understanding the "worried well". PMID- 11927061 TI - Primary data for primary care. PMID- 11927060 TI - The probability of specific diagnoses for patients presenting with common symptoms to Dutch family physicians. AB - OBJECTIVE: Our goal was to develop reliable data on the probability of specific diagnoses among patients of family physicians (FPs) presenting with common symptoms. STUDY DESIGN: A group of 54 Dutch FPs recorded the reasons for encounter, diagnoses, and interventions for all episodes of care between 1985 and 1995. Diagnoses could be modified during the episode of care, and a modified diagnosis was applied to all episode data. POPULATION: We used the listed patient populations of the 54 Dutch FPs, representing 93,297 patient years, 236,027 episodes of care, and 267,897 patient encounters. OUTCOMES MEASURED: The top 20 diagnoses related to 4 selected presenting symptoms (cough, shortness of breath, general weakness/tiredness, and low back symptom/complaints without radiation), per 100 patients, with 95% confidence intervals, stratified by age. In the standard tables, age-specific cells with fewer than 10 observations were excluded. RESULTS: The availability of an accurate estimate of prior (pretest) probabilities for common symptoms/complaints has great potential value for family practice as an academic discipline and for family physicians in that it can support their medical decision making. Stratifying data by age groups increases the clinical relevance of the prior probabilities. CONCLUSIONS: Though collected by Dutch FPs, the data in our study have a high face validity for other clinicians. Still, FPs in other countries should give priority to collecting their own probability databases. PMID- 11927062 TI - The effectiveness of magnet therapy for treatment of wrist pain attributed to carpal tunnel syndrome. AB - We conducted a double-blind placebo-controlled randomized clinical trial in which 30 patients with pain attributed to carpal tunnel syndrome had either a 1000 gauss magnet or a placebo metal disk applied to the carpal tunnel area using a Velcro wrap for a period of 45 minutes. Pain was measured on a visual analogue scale using 0 and 10 as anchors. Presenting symptoms including numbness, tingling, burning, and pain did not differ significantly between the 2 groups. There was significant pain reduction across the 45-minute period for both groups. However, t test comparisons found no significant differences between the groups for beginning pain, pain at 15 minutes, pain at 30 minutes, or pain at 45 minutes. The use of a magnet for reducing pain attributed to carpal tunnel syndrome was no more effective than use of the placebo device. PMID- 11927063 TI - Screening for alcoholism in the primary care setting: are we talking to the right people? AB - BACKGROUND: This study assessed which demographic groups were most likely to consume alcohol excessively, and which groups had received inquiries and discussion about alcohol use from their physicians compared with discussions about other health risks. STUDY DESIGN: This was a cross-sectional study using data from the Centers for Disease Control Behavioral Risk Factors Surveillance System 1997 data set that represents a stratified random sample in the United States. POPULATION: We selected 23,349 adults who reported a routine physical examination within the last 3 years. OUTCOMES MEASURED: The main variables involved responses to questions about alcohol intake and whether the respondent's physician had initiated discussions about drinking. RESULTS: Physicians spoke to patients about alcohol use much less frequently than about other health-related behaviors. Discussions were roughly targeted to groups with the largest intake. However, physicians were least likely to speak with white patients, women, and widows who drank significantly. CONCLUSIONS: Regularly asking patients about alcohol use could substantially reduce the under-recognition of alcoholism. Since brief counseling is effective, negative consequences of excessive alcohol intake may be avoided. PMID- 11927064 TI - Do parents and physicians differ in making decisions about acute otitis media? AB - OBJECTIVES: We wanted to discover how parents differ from physicians in making decisions about how to treat a child who may have acute otitis media (AOM). STUDY DESIGN: We used questionnaires that required participants to judge the probability of AOM or choose treatment for 2 sets of 46 paper scenarios of hypothetical children aged 15 months who might have AOM, and they subsequently rated the importance of individual cues and described their attitudes and opinions related to health care and AOM. POPULATION: Convenience samples of 19 US family physicians, 35 French generalists, 21 French pediatricians, 52 US parents, and 86 French parents were included. OUTCOMES MEASURED: The primary outcomes were the judgment policies-the weights placed on each of the scenario cues when making decisions-that were derived for each individual and each group by multiple linear regression. RESULTS: The mean judged probabilities of AOM were nearly the same for all groups: 50% for the US physicians, 51% for the US parents, 52.5% for the French physicians, and 52% for the French parents. The percentages of cases treated with antibiotics did not differ: 53% for US physicians, 45% for US parents, 53% for French physicians, and 51% for French parents. All groups gave greatest weight to the physical examination cues for decisions about both diagnosis and treatment. The parents paid little attention to the cues that reflected parental concerns. CONCLUSIONS: US and French parents were very similar to physicians in their judgments and treatment choices regarding AOM. They appear to be able to adopt the physician's point of view and to be selective in the use of antibiotics. PMID- 11927065 TI - Shoulder joint capsule distension (hydroplasty): a case series of patients with "frozen shoulders" treated in a primary care office. AB - "Frozen shoulder," most often caused by adhesive capsulitis, is frequently treated with intra-articular steroid injections, physical therapy, and surgical manipulation under anesthesia. These therapies provide limited benefits. Hydraulic distension of the shoulder joint capsule (hydroplasty) has potential to provide rapid relief of pain and immediate improvement of shoulder function for patients with adhesive capsulitis. We performed 21 hydroplasty procedures on 16 patients over a 4-year period. Ninety-four percent (17/18) of the procedures improved patient's measured mobility immediately after the procedure. Fifty-three percent (10/19) of the procedures produced immediate, short term, and sustained improvement in comfort and function. No significant complications of the procedure were detected. Our series suggests that the hydroplasty procedure should be further evalutated. PMID- 11927066 TI - Tobacco cessation counseling among underserved patients: a report from CaReNet. AB - OBJECTIVES: The purpose of our study was to determine the frequency of smoking cessation counseling in relation to insurance status in a practice-based research network. STUDY DESIGN: We administered a modified National Ambulatory Medical Care Survey (NAMCS), with an additional payment category to identify uninsured patients, quarterly to 100 random patients at each practice site for 1 year. POPULATION: The study population included the patients at the 7 practices within the Colorado Research Network (CaReNet), associated with the Department of Family Medicine, University of Colorado Health Science Center. OUTCOMES MEASURED: We measured the prevalence of smoking and the frequency of cessation counseling. RESULTS: Of 2773 visits analyzed, 1443 were made by adults who were either was uninsured (39%), had Medicaid (22%), or had private or a health maintenance organization insurance (private/HMO; 40%). Smoking prevalence was significantly greater in uninsured patients (30%) and Medicaid patients (31%), compared with private/HMO patients (22%) (P =.008). However, those smokers with private/HMO insurance were more likely to receive tobacco counseling (50%) than Medicaid (41%) and uninsured (25%) patients (P <.001). After controlling for potential confounders, this difference remained significant. CONCLUSIONS: Although smoking is more common among Medicaid and uninsured patients, these smokers are less likely to receive counseling. Possible explanations for this disparity include lack of access to cessation interventions or lower quality of care for underserved patients. This finding may have implications for achieving national public health goals on smoking cessation. PMID- 11927067 TI - Physician recruitment for a community-based smoking cessation intervention. AB - OBJECTIVE: Our goal was to describe a strategy to recruit a population-based sample of physicians into a trial to test an approach to disseminate physician delivered smoking cessation interventions. STUDY DESIGN: The 3-phase population based recruitment trial included: (1) a print-based promotional appeal, (2) in person presentations with by the principal investigator (PI), and (3) follow-up calls by the PI and paid physician recruiters. Participation requirements were kept minimal to facilitate recruitment. POPULATION: All primary care physicians statewide were targeted; 3 counties were chosen as intervention areas and 2 counties as control areas. A subsample of physicians was targeted in the larger control areas through a matching process. OUTCOME MEASURED: We measured physician recruitment rate. RESULTS: Eighty-one percent (n=259) of all eligible physicians were successfully recruited into our study. CONCLUSIONS: The full multistep process was important in getting participation agreement. By using an intensive recruitment strategy and minimizing research demands, it is possible to recruit community-based primary care physicians for research projects that will help them enhance the preventive services they provide to their patients. PMID- 11927068 TI - The role of family practice in different health care systems: a comparison of reasons for encounter, diagnoses, and interventions in primary care populations in the Netherlands, Japan, Poland, and the United States. PMID- 11927070 TI - Cigarette smoking and erectile dysfunction. PMID- 11927069 TI - The treatment of adults with essential hypertension. AB - Hypertension is arbitrarily defined as diastolic blood pressure (DBP) of 90 mm Hg or higher, systolic blood pressure (SBP) of 140 mm Hg or higher, or both, on 3 separate occasions. Essential hypertension is hypertension without an identifiable cause. Essential hypertension, also known as primary or idiopathic hypertension, accounts for at least 95% of all cases of hypertension. According to the third National Health and Nutrition Examination Survey (NHANES III), approximately 60% of the 50 million Americans with hypertension are at increased risk for cardiovascular disease resulting from uncontrolled hypertension. This is because only 53% of hypertensive patients are being treated and only 24% have their hypertension under control. Physicians must play an active role in identifying and treating hypertension. In an earlier Applied Evidence article, an approach to the diagnosis of hypertension was presented. This article reviews the treatment of essential hypertension in adults and the prognosis of untreated hypertension. Risk stratification, alternative therapies, lifestyle modification, drug therapy, and prognosis will each be reviewed sequentially. PMID- 11927071 TI - Infant circumcision. PMID- 11927072 TI - Are once-daily iron drops as effective as thrice-daily therapy in children with iron deficiency anemia? PMID- 11927073 TI - What is the diagnostic accuracy of the clinical examination for meniscus or ligamentous knee injuries? PMID- 11927074 TI - Are oral contraceptives (OCPs) with anti-androgenic progestins preferred over other OCPs in patients with acne? PMID- 11927075 TI - Is oral oseltamivir safe and effective for the prevention of influenza and its complications in frail elderly long-term care residents who have received influenza vaccine? PMID- 11927076 TI - [Health effects of the Chernobyl disaster. Fifteen years afterwards]. AB - The Chernobyl nuclear accident of 1986 released large quantities of radioactive material causing heavy contamination in widespread areas of the former Soviet Union. Each summer, several hundred children visit Spain from Chernobyl. In this article we describe the accident, the environmental contamination, the mechanisms of radiation injury and the dose-response relationships. We review the health effects of exposure to ionizing radiation and the health impact of the Chernobyl catastrophe. We propose guidelines for the medical management and evaluation of children on temporary visits. The health status of adults and especially that of children in Belarus, Ukraine and the Russian Federation has been adversely affected. According to present knowledge, Chernobyl has given rise to a marked increase in the incidence of papillary thyroid cancer, psychological consequences and socioeconomic disruption. Many studies report that the incidence of other diseases has increased, but not all health problems seen after the nuclear accident can be attributed to radiation. Given the long latency period for diseases induced by radiation exposure, long-term follow-up of all potentially affected individuals is important. Fifteen years after the Chernobyl accident the international community is still learning scientific, medical and humanitarian lessons. PMID- 11927077 TI - [Guidelines for respiratory syncytial virus prophylaxis. An update]. AB - Epidemiological studies performed by the IRIS study group in the last two respiratory syncytial virus (RSV) seasons found that the hospitalization rates for RSV in premature infants born before or in week 32 of gestation were 13.4 % and 13.1 %, respectively. Of these, 18 % and 25 % of the infants were admitted to the intensive care unit. Currently available information demonstrates the efficacy of RSV monoclonal antibodies (palivizumab) and the absence of major adverse effects. To date, there are no data that indicate the need to modify the guidelines for RSV prophylaxis in premature infants published in 2000. PMID- 11927078 TI - [Neonatal onset methylmalonic aciduria and homocystinuria:Biochemical and clinical improvement with betaine therapy]. AB - Methylmalonic aciduria and homocystinuria is a very rare inborn error of cellular cobalamin (Cbl) metabolism. We describe the biochemical evolution and clinical course of a boy with neonatal onset CblC mutant defect.Born after a normal pregnancy, the patient developed general hypotonia and severe feeding difficulties at 5 days of life. Diagnosis of methylmalonic aciduria and homocystinuria was established by amino-acid and organic acid analysis and was confirmed by enzyme and genetic studies. The patient was initially treated with parenteral hydroxocobalamin (1 mg/day), oral carnitine (100 mg/kg/day) and a restricted protein diet. This treatment returned methylmalonic acid levels to normal. Despite the parenteral hydroxocobalamin therapy, the patient showed no improvement in neurological dysfunction, hypotonia or developmental delay. Oral betaine supplementation (3 g/day) from months 3-15 reduced plasma total homocysteine and homocystinuria. The patient showed clinical improvement in neurological and growth development. We conclude that early betaine therapy was safe and effective in our patient with neonatal onset methylmalonic aciduria and homocystinuria type CblC. PMID- 11927079 TI - [Crouzon's syndrome with acanthosis nigricans]. AB - Crouzon's syndrome is a complex craniosynostosis disorder due to mutations in fibroblast growth factor receptor (FGFR) type 2. We report a female patient with Crouzon's syndrome associated with acanthosis nigricans. The molecular abnormality in this patient is a point mutation (Ala391Glu) in the transmembrane domain of another FGFR (type 3), which is very close to the mutation (Gly380Arg) most frequently observed in achondroplasia. Acanthosis nigricans is an emerging disorder. Its clinical features and molecular findings differ from those of isolated Crouzon's syndrome. These data are very useful when molecular tests are required for prenatal diagnosis. PMID- 11927080 TI - [Deletion of thymine at position 2298 in exon 5 of the androgenic receptor gene causing complete androgen insensitivity syndrome]. AB - BACKGROUND: Androgen insensitivity syndrome is an X-linked disorder of male sexual differentiation caused by mutations in the androgen receptor gene and resulting in a wide range of phenotypes. OBJECTIVE: To study the androgen receptor gene in two cousins with androgen insensitivity syndrome. PATIENTS AND METHODS: We present two patients who attended our clinic for primary amenorrhea. The phenotype and external genitalia were female. Pelvic ultrasonography showed the absence of uterus and female internal genitalia. In both patients the karyotype was 46 XY and consequently both patients underwent bilateral gonadectomy. Histological examination confirmed that the gonads were testes. Molecular study of the androgen receptor gene was performed to confirm androgen insensitivity syndrome. RESULTS: Both patients showed a thymine deletion in exon 5 at nucleotide 2298 (codon CCT for proline 766) of the androgen receptor gene, causing their phenotype. CONCLUSIONS: To confirm androgen insensitivity syndrome,t he androgen receptor gene should be analyzed for mutations, although the relationship between genotype and phenotype is weak. To detect carriers of the mutation, karyotyping and study of the androgen receptor gene should be performed in girls who are first relative of the probands. PMID- 11927081 TI - [Steatosis and cirrhosis secondary to insulin resistance in children. Possible physiopathological mechanisms]. AB - We describe two girls who showed physical and biochemical features of insulin resistance and steatohepatitis. The possible pathophysiological mechanisms involved in fatty liver and hepatotoxicity are discussed. PMID- 11927082 TI - [Nontuberculous bacterial infection in immunocompetent children]. AB - Nontuberculous or environmental mycobacterial disease in children has been increasingly recognized over the last decade. We present four patients who were diagnosed in the year 2000. The children were aged between 2 and 8 years. Three patients presented involvement of the cervical lymph nodes and one presented involvement of the inguinal nodes. Three of the children were treated with a combination of surgery and chemotherapy and one was treated with chemotherapy alone. We describe the clinical characteristics, laboratory findings, therapeutic management and complications of nontuberculous mycobacterial lymphadenitis in children. PMID- 11927083 TI - [Laryngotracheal trauma]. PMID- 11927084 TI - [Nebulized epinephrine in acute bronchiolitis. Is there enough evidence?]. PMID- 11927085 TI - [Bronchiolitis and epinephrine: Reviewing the evidence]. PMID- 11927086 TI - [Incidence, microbial etiology and mortality associated with nosocomial bacteremia in a neonatal intensive care unit]. PMID- 11927087 TI - [Meningitis as a neurological complication of herpes zoster]. PMID- 11927088 TI - [Schistosoma haematobium as a cause of hematuria]. PMID- 11927089 TI - [Postpericardiotomy syndrome: Late recurrences]. PMID- 11927090 TI - [Toxic epidermal necrolysis treated with intravenous immunoglobulin]. PMID- 11927091 TI - [Pyriform sinus fistula: An unusual cause of recurrent suppurative thyroiditis]. PMID- 11927092 TI - [Continuous splenic-gonadal fusion]. PMID- 11927093 TI - [Newborn with asymmetric face and heart murmur]. PMID- 11927094 TI - [Prophylaxis of respiratory syncytial virus infections with palivizumab]. PMID- 11927095 TI - [Chemoprophylaxis with palivizumab in Andalusia (Spain). Results of the 2000-2001 respiratory syncytial virus epidemic]. AB - OBJECTIVE: To assess the hospitalization rate for bronchiolitis in newborns treated with palivizumab in Andalusia. PATIENTS AND METHODS: We performed a prospective study of 283 neonates and infants who received prophylaxis with palivizumab in Andalusia during the 2000-2001 bronchiolitis epidemic. We also performed a descriptive study of hospitalization for bronchiolitis, classifying patients according to gestational age and whether they had bronchopulmonary dysplasia (BPD). RESULTS: Most (86.9 %) of the treated patients were born before week 32 of gestation (63 % before week 30) and 38.4 % developed BPD. A total of 10.6 % of palivizumab-treated newborns were hospitalized for bronchiolitis, of which 3.9 % was caused by respiratory syncytial virus (RSV) (4.8 % of patients with BDP and 3.5 % of those without BPD were RSV-positive). Compared with preterm neonates born in weeks 31 or 32 of gestation, palivizumab-treated newborns born before week 31 showed a higher hospitalization rate for bronchiolitis (13.3 % vs 6.3 %; p < 0.05) and that for RSV was three times higher (5.0 % vs 1.6 %; p < 0.05). Age at admission was 5.8 3.2 months. Length of hospital stay was shorter in RSV-positive patients with bronchiolitis (6.7 vs 7.6 days; p > 0.10). Admission to the intensive care unit was required in 0.7 % of patients and in 50 % of those who were RSV-positive. Adverse effects were observed in 5.1 % of palivizumab-treated patients. CONCLUSION: The marked differences in the perinatal antecedents of patients receiving immunoprophylaxis may explain the variations in the efficacy of palivizumab reported in available studies. PMID- 11927096 TI - [Epidemiological features of exercise-induced bronchial hyperresponsiveness in children aged 13-14 years old in Barcelona (Spain)]. AB - BACKGROUND: Conclusive data on the risk factors that modify bronchial hyperresponsiveness after airway exposure to modulating triggers (histamine, methacholine and exercise) are lacking. In recent years some risk factors such as air pollution or infection have been questioned and even considered protective. OBJECTIVES: To assess the prevalence of bronchial hyperresponsiveness to exercise among schoolchildren and its association with different risk factors. METHODS: Three thousand thirty-three schoolchildren aged 13-14 years were studied. Participants answered a questionnaire on respiratory symptoms. A total of 2,842 children underwent exercise challenge testing for bronchial hyperresponsiveness. This study was part of the International Study of Asthma and Allergies in Childhood (ISAAC). RESULTS: A fall in peak expiratory flow rate of (15 % after exercise was found in 324 children (11.4 %), of whom 29 (9 %) also presented symptoms of asthma. Bronchial hyperresponsiveness to exercise was significantly associated with lower age, female sex, high socioeconomic level and attending a private school. No association was found between bronchial hyperresponsiveness and obesity, tobacco or viral infections. CONCLUSIONS: These findings are not completely consistent with those of other studies, suggesting that local environmental factors may influence the prevalence of bronchial hyperresponsiveness. Further studies are required to clarify these conflicting data. PMID- 11927097 TI - [Agreement between tracheal auscultation and pulmonary function in methacholine bronchial inhalation challenge in asthmatic children]. AB - BACKGROUND: PC wheezing (PCw) is defined as the concentration of methacholine at which wheeze is detected on auscultation of the trachea. PCw has been suggested as a measure of bronchial hyperresponsiveness in methacholine challenge testing (MCT). OBJECTIVE: The aim of this study was to determine the agreement between the concentration of methacholine that produces a 20 % decrease in forced expiratory volume in 1 second (FEV1) (PC20) and PCw in MCT in asthmatic children. PATIENTS AND METHODS: Eighteen asthmatic children with a mean age of 11.5 years (range: 6-16 years) were studied. Fifteen of the children were under treatment with inhaled glucocorticoids. MCT was performed according to the guidelines of the American Thoracic Society (1999) using a Hudson nebulizer calibrated to obtain a mean output of 0.14 ml/min. After each nebulization, two independent observers registered FEV1 and tracheal auscultation. FEV1 was determined by forced spirometry 30 and 90 seconds after the end of nebulization and PC20 was registered (exponential model). Respiratory rate and transcutaneous oxygen saturation were continuously monitored. Tracheal auscultation was performed at 0, 60 and 120 seconds after the end of nebulization. The end point was defined as the appearance of wheezing over the trachea. The values of PC20 and PCw, as well as the concentration of methacholine corresponding to a decrease in FEV1 equal to or higher than 20 %, were compared using Student's matched pairs-test and Wilcoxon's test. The degree of agreement between variables was compared by using Bland-Altman's test. RESULTS: MCT was positive in 17 of 18 patients. No differences were found between PC20 and PCw (p 0.15). Both variables showed agreement in 12 of 17. A clear association was found between both measures (log PCw, log PC20): R: 0.92; p < 0.001. The mean decrease in FEV1 on reaching PCw was 24.8 % (range: 10-41). No adverse effects were observed. CONCLUSION: The agreement between PC20 and PCw in MCT in asthmatic children is excellent. PCw could be helpful in determining bronchial hyperresponsiveness in young asthmatic children in whom spirometry is not feasible. PMID- 11927098 TI - [Tonsillar and erythrocyte levels of superoxide dismutase in children with and without recurrent tonsillitis]. AB - BACKGROUND AND OBJECTIVES: Oxidative stress secondary to infant tonsillar infection produces the expression of local and systemic antioxidants. Its determination seems to be useful as a marker of tonsillar suffering before tonsillectomy but is currently unknown. The aim of this study was to determine the evolution of this parameter in tonsillectomized children during a long-term follow-up. METHODS: One hundred thirty-six children underwent tonsillectomy, 46 for tonsillar hypertrophy without infection and 90 for recurrent tonsillitis. Superoxide dismutase (SOD) concentrations were measured before tonsillectomy and at 6, 12, 24 and 36 months post-tonsillectomy. RESULTS: Infection provoked significantly higher SOD concentrations than tonsillar hypertrophy in tonsillar tissue (223.06 30.46 vs 156.39 54.05 U/l, p < 0.001) and in blood (1124.91 141.73 vs 1007.19 97.03 U/gr Hb, p < 0.001). There was a strong correlation between tonsillar and erythrocyte concentrations. During the 3-year follow-up, SOD concentrations in blood progressively decreased until stabilizing in all patients. Stabilization was reached at 6 months post-tonsillectomy in the group with tonsillar hypertrophy and at 2 years in the group with infection. Children with recurrent tonsillitis consistently showed higher SOD concentrations in blood throughout the follow-up. CONCLUSIONS: Oxidative damage in tonsillar tissue results from the incidence and severity of focal infections. Tonsillectomy reduces SOD levels but, as a consequence of oxidative stress, these do not return to normal. PMID- 11927099 TI - [Antipyretic poisoning]. AB - OBJECTIVE: To establish the current incidence of poisoning caused by oral antipyretics in the Spanish pediatric population. METHODS: From January 1998 to December 2000, all cases of poisoning due to antipyretic ingestion in children aged up to 14 years old were recorded and tabulated in the Poison Control Center. RESULTS: A total of 13,044 cases of drug poisoning were recorded. Acetaminophen accounted for 11.0 %, acetylsalicylic acid (ASA) for 3 % and ibuprofen for 1.5 % of the cases (p < 0.001). The risk of acetaminophen poisoning was 5.6 higher than that of ibuprofen poisoning (RR: 5.6; 95 % CI: 4.8-6.5). Seventy three percent of poisonings occurred in children aged 1-3 years old. Poisonings were considered serious in 9.4 % of those produced by acetaminophen, 2 % of those produced by ASA and 1 % of those produced by ibuprofen (p < 0.001). Information from the Poison Control Center were requested mainly by emergency department physicians (78.9 %), followed by pediatricians (15.4 %), other physicians (4.7 %) and nursing staff (1 %). CONCLUSIONS: Because of their frequency and potential toxicity, drug and household product poisonings in childhood are a significant public health problem. Among the antipyretics evaluated, acetaminophen was the most frequently associated with poisoning and with severe intoxications. We recommend several preventive strategies to reduce the incidence of drug poisoning in childhood. PMID- 11927100 TI - [Catalan version of Barcelona Test's verbal subtests]. AB - Test Barcelona. Bilinguismo catalan-castellano. Adaptacion linguistica de instrumentos neuropsicologicos. BACKGROUND AND OBJECTIVES: The present paper develops the Catalan version of the language subtests of the Barcelona Test (Pena Casanova,1986) the original version of which was published in Spanish. METHODS: To reach the Catalan version of the test it has been necessary to analyse the psycholinguistic variables of every item to then establish linguistically equivalent correlations. RESULTS: There are some subtests for which no modifications have been needed. Nevertheless, and because of some psycholinguistic features of Catalan (Catalan's spelling is not completely free of ambiguity) some subtests have suffered substantial modifications. The following paper presents the justification of every modification based on the features of the Catalan language and also based on the original goals of the test. An appendix contains the final version of the Catalan adaptation for the language subtests of the Barcelona Test. CONCLUSION: The present paper represents an advance in the assessment of neurological patients with language disfunctions. It will permit an appropiate assessment for the Spanish-Catalan bilingual patients. PMID- 11927101 TI - [Hypomethylation and multiple sclerosis, the susceptibility factor?]. AB - BACKGROUND: Azathioprine, off-label used long time ago to treat multiple sclerosis (MS) patients, has recently received approval from the Spanish Medicine Agency (Agencia Espanola del Medicamento) in relapsing-remitting (RR) forms of this condition. Clinical efficacy of azathioprine is due to the enzymatic conversion to 6-thioguanine (the active metabolite). The key enzyme in this process is thio purine methyl transferase (TPMT), converting 6-MP to 6-methylMP. A specific genetic polymorphism has been described affecting this enzyme. With the aim of optimizing purine therapy in a variety of autoimmune diseases, monitoring of TPMT phenotype has been performed in a vast number of patients. The TPMT activity frequency distribution histogram from a Spanish population sample has been compared with the corresponding ones to Crohn's disease, ulcerative colitis and MS patients. METHODS AND RESULTS: TPMT activity has been studied in red blood cells obtained from 3,640 clinical laboratory samples in Spain of which 1,249 corresponded to patients affected by Crohn's disease, 589 to ulcerative colitis, 348 to MS, 487 to several autoimmune diseases apart from the previously mentioned and 967 to a group of blood donors. The mean TPMT activity in the MS group (17.1 6.1 U/ml) was significantly lower (p < 0.001) than in Crohn's disease (20.0 5.8 U/ml), ulcerative colitis (19.7 6.1 U/ml) and donors group (19.9 6.3 U/ml). CONCLUSION: Defective methylation profile and subsequent hyperhomocysteinemia leading to a widespread impairment of the methyl-transferase activity (in this case affecting MBP methylation) is a vicious circle we propose as a MS susceptibility factor. PMID- 11927102 TI - [Lamotrigine in the adult-onset epilepsy: efficacy and long-term safety]. AB - BACKGROUND: LTG is a new antiepileptic drug that is nowadays very often used in epileptic patients. OBJECTIVES: To determine efficacy and safety of Lamotrigine (LTG) in the first five years after its marketing in patients at a third level university hospital, as well as its impact on the management of classic antiepileptic drugs (AED). PATIENTS AND METHOD: We reviewed retrospectively our Epilepsy Unit Database. One hundred patients were treated with LTG in a 5-year period. Efficacy was evaluated comparing seizure frequency in a 6-month period before and after LTG. The type of epilepsy, side effects, blood levels and concomitant treatments were considered in the analysis. RESULTS: LTG was effective in all groups of epileptic patients studied. Eighteen percent of patients became seizure-free. Seventeen percent of patients improved more than 50%. Fifty-seven percent of patients remained treated with LTG after four years of follow-up. Side effects were mild, but frequent; only four patients discontinued LTG because of adverse effects. Serum levels were usually high, but showed no relation with clinical efficacy. The mean number of AED taken per patient increased. CONCLUSIONS: LTG is a safe an effective drug in epilepsy. It has a clear impact in the management of the epileptic patients. PMID- 11927104 TI - [The turn of the screw: complex visual hallucinations in the Henry James' novel]. AB - The turn of the screw is one of the most celebrated stories by Henry James. It is also a top writing within the so-called fantastic literature, whose narrative strength comes from the intermittent visions suffered by the main character. The vividness and dramatic content that represent the firstly unidentified human figures, that moreover recur as brief, stereotyped and fragmentary images, are constitutive of complex visual hallucinations. These characteristics, alongside acute premonitory symptoms such as emotional changes (fear, anxiety) or altered thinking (forced, "deja vu", "jamais vu"), and the final altered awareness or loss of consciousness, allow us to infer an epileptic nature of the ten episodes described. Postictal psychosis, that follows a lucid interval and may last up to the several weeks encompassed by the story, would account for the paranoia featured, in the setting of a temporal lobe epilepsy. The accurate descriptions prompted us to search for autobiographical, scientific or literary influences: The alcoholism and visual hallucinations suffered by his father, the knowledge on hallucinations provided by his brother Williams on his paramount and former The Principles of Psychology, and an early devotion to Poe's writings, an epileptic himself with excellent descriptions of seizures in his writings, might have enabled the author to perform his story with such a hallmark of neurological details. PMID- 11927105 TI - [Paraneoplastic retinopathy and anti-recoverin antibodies]. PMID- 11927103 TI - [Lhermitte's sign]. AB - Besides Babinski's, Lhermitte's sign is likely the eponym mostly used in the neurological literature. We review here the history of this eponym as well as recent advances on its pathophysiology and treatment. Lhermitte's phenomenon is, on one hand, a symptom as it is spontaneously explained by patients, and, on the other hand, a sign as it may be triggered by flexion of the nape. Initially described after head and cervical spine trauma, firstly by Marie and then by Babinski, it was Jean Lhermitte who recognized on it an etiological specificity, namely, a demyelinating sign of cervical spinal posterior cords. He also made a pathophysiological interpretation of the phenomenon, namely, a stretching of posterior cords during flexion of the neck. All authors agree that this phenomenon is more common in multiple sclerosis, although it has been descibed in many other conditions. The history of how this sign was hatched, whose pathophysiology remains a mystery, is fascinating. And it is fascinating the fact that patients compare the phenomenon with a current, especially if we bear in mind that few people may have suffered an electrocution, mainly in that time when just a few ones could actually enjoy domestic electricity. PMID- 11927106 TI - [Adult GM2 gangliosidosis: improvement of ataxia with GABAergic drugs]. AB - The authors present a case of adult GM2 gangliosidosis, B1 enzymatic type. The main clinical features found were cerebellar ataxia, proximal lower limb weakness and myokymia. The neurological examination, and the biochemical, electrophysiologic and imaging studies are all described. Decreased activity of the enzyme beta-hexosaminidase A in the metabolism of the sulfate substrate 4-MU NAGS was found in serum. Global cerebellar atrophy was observed in a cranial nuclear magnetic resonance. The electrophysiologic study showed continuous spontaneous activity integrated by myokymia and neuromyotonic discharges in addition to signs of acute and chronic denervation. Disappearance of the myokymia and improvement in the ataxia were attained with the use of the GABAergic drugs gabapentin and tiagabine. The authors try to explain the clinical improvement obtained with the drugs by relating their mechanisms of action to the central nervous system neurotransmitter alterations proposed for this disease. PMID- 11927107 TI - Myopathic camptocormia in a patient with levodopa unresponsive parkinsonism. AB - Camptocormia may be seen in Parkinson's disease. As no changes in paraspinal musculature are found, it is attributed to dystonia or extreme rigidity. However, several cases of parkinsonism and dropped head due to neck extensor myopathy have been reported. We report the first patient with levodopa unresponsive parkinsonism and camptocormia of muscular origin. PMID- 11927108 TI - [Hydrosyringomyelia in demyelinating diseases]. AB - Spinal cord cavitation is a frequent finding in optic neuromyelitis (Devic's syndrome) (DS) but it is also, although rarely, observed in patients with multiple sclerosis (MS). The objective of our study was to compare the MRI characteristics of the syringomyelic cavities in 6 patients with DS and 3 patients with MS. All the patients with DS had a relapsing clinical form with normal brain MRI. Spinal MRI revealed unenhanced central cavities which extended more than 3 vertebral bodies and remained unchanged in follow-up studies. Two patients presented multiple cavities.MS patients suffered a relapsing remitting form of the disease, they all had hyperintense T2 enhancing lesions on their spinal MRI. Moreover spinal MRI also revealed non communicating cavities which extended less than 2 vertebral bodies. Follow-up studies in MS patients revealed a reduction of both the spinal lesions and the cavities. It is still debated whether DS represents a distinct clinical entity different from MS. These findings help distinguishing both disorders in cases when spinal cavities are present and also contribute to the therapeutic choice. PMID- 11927109 TI - [Intracranial hypertension associated with type I Chiari malformation]. PMID- 11927110 TI - [Polyneuropathy associated with monoclonal gammapathy of unknown origin: report of a case]. PMID- 11927111 TI - [Fulminant coma as the first manifestation of organophosphate poisoning]. PMID- 11927112 TI - [Declaration of conflicts of interests]. PMID- 11927114 TI - Effects of the Yuzpe regimen, given during the follicular phase, on ovarian function. AB - This study was conducted to assess to what extent the Yuzpe regimen, or half the dose, given in the follicular phase, prevents ovulation during the ensuing 5 days. Sixty women were divided into six groups. All groups received placebo in one cycle and drug in another, in a randomized order. Groups differed by the dose and size of the leading follicle at the time of treatment (12-14, 15-17, or 18-20 mm). Ovulation was absent during the ensuing 5 days in 13 of 20 participants (65%) and in 8 of 20 participants (40%) who received the full and the half dose, respectively, when follicles were 12-17 mm. No ovulation occurred, within the critical period, in 7 of 39 placebo cycles (18%). When follicles were 18-20 mm, treatment did not prevent ovulation. In most drug-treated cycles, plasma gonadotropin and sex steroid levels were significantly depressed within the 5-day period, even when follicular rupture occurred within that period. In conclusion, the Yuzpe regimen can suppress or postpone ovulation to an extent that exceeds the fertile life of spermatozoa. Lack of ovulation within the critical period and dysfunction of the ovulatory process probably account for the contraceptive effect of this method in most cases. The present data do not warrant the use of half the dose of the Yuzpe regimen. PMID- 11927115 TI - Bleeding patterns and clinical performance of the levonorgestrel-releasing intrauterine system (Mirena) up to two years. AB - The objectives of this study were to evaluate the bleeding patterns and clinical performance during the first 2 years of use of an intrauterine system releasing 20 microg/day of levonorgestrel (LNG-IUS, Mirena). Two-hundred-fifty-six women accepted use of Mirena from April 1998 through September 1998. The gross cumulative discontinuation rate due to pregnancy and expulsion were significantly higher in women who used the device because of heavy bleeding. There was one pregnancy at the 15th month of use after an inadvertent expulsion of the device. The continuation rate was 66.2 at the end of the second year. Forty-four percent of women reported amenorrhea at the 6th month of use. This rate maintained stability at 50% after 12 and 24 months of use. Spotting was present in 25% of the users at 6 months, decreasing to 8% and 11% at 18 and 24 months, respectively. Oligomenorrhea was described by one-quarter of women and was similar at each observation period. Removals due to menstrual bleeding problems were concentrated in the first 6 months of use and mostly due to amenorrhea or menorrhagia. In conclusion, LNG-IUS showed a high contraceptive efficacy and a good continuation rate up to 2 years. In addition, a reduction of blood loss was observed a few months after insertion. PMID- 11927116 TI - Could American women use mifepristone-misoprostol pills safely with less medical supervision? AB - Medical methods of early abortion differ from surgical methods in that women themselves can potentially administer the regimens. As currently researched and offered, however, the main regimen used for medical abortion, mifepristone misoprostol, is highly medicalized, involving several clinic visits and extensive physician involvement. We re-examined the role of clinical supervision in each step of the abortion process, using data collected during a large clinical trial of mifepristone-misoprostol abortions in the US, fielded during 1994-1995. The trial was carried out in 17 geographically diverse centers, including private, public, and nongovernmental organization clinics, and enrolled 2121 women, aged 18-45 years, seeking early abortion (< or =63 days since last menstrual period). Women received 600 mg oral mifepristone, followed 48 h later by 400 microg oral misoprostol. Evidence suggests that most women can handle most steps of the medical abortion process themselves, effectively and safely. The utility of clinic visits to ingest mifepristone and misoprostol is questionable. For many women, even the follow-up visit could perhaps be replaced by telephone follow-up, combined with home pregnancy tests. Alternatives to the present protocol might allow greater control, comfort, and convenience at lower cost. Where clinician involvement might be useful, mid-level health care providers typically possess the skills necessary to offer the method safely, implying that physicians might be necessary only as complications arise. Future research useful for determining the optimal amount of medical involvement to provide mifepristone-misoprostol safely and effectively should include self-screening tests, label comprehension tests, calendars to aid in calculating gestational age, and the development of special pregnancy tests with telephone follow-up. PMID- 11927117 TI - Policy climate, scholarship, and provision of emergency contraception at affiliates of the International Planned Parenthood Federation in Latin America and the Caribbean. AB - Emergency contraception (EC) has great potential to decrease the incidence and resulting consequences of unwanted pregnancy, including unsafe abortion. We conducted this study to understand EC practices in Latin America and the Caribbean (LAC). We contacted 43 International Planned Parenthood Federation affiliates in LAC to interview them about EC availability. We collected family planning norms and researched registered EC products in LAC. We searched English- and Spanish-language sources to compile EC literature reviews. Thirty-seven affiliates (86%) responded to the survey, and 62% offer EC. Central and South American affiliates are more likely to offer EC than are Caribbean affiliates. Of those offering EC, 96% offer cut-up packets of oral contraceptives, whereas six affiliates offer dedicated products. Of those not offering EC, 79% believe it constitutes abortion. EC availability and support for the method appear to be increasing in LAC, and clearer distinctions between EC and abortion in medical and policy guidelines should increase acceptance further. PMID- 11927118 TI - Predictors of difficulty inserting the female condom. AB - This article describes the frequency of initial difficulty inserting the female condom and identifies predictors of insertion difficulty among women at risk of sexually transmitted diseases (STDs). Female STD clinic patients (n = 1144) were taught how to insert the female condom by using an anatomic model, then given an opportunity for self-insertion practice. Correct placement of the condom was verified by a nurse clinician, and the number of attempts required for correct insertion was recorded. Sociodemographic and psychosocial predictors of refusing the insertion practice and of difficulty inserting the female condom were evaluated using logistic regression. Only 5% of study participants refused the self-insertion practice. Women who never had a Papanicolaou smear test, did not use tampons, never used an inserted method of STD prevention/birth control, and disliked the insertion features of intravaginal barrier methods were more likely to refuse the self-insertion practice. Of those who attempted self-insertion, 25% were unable to insert the female condom correctly on the first attempt. Women who never expressed their sexual likes and were indifferent to the positive features of intravaginal contraceptive methods were more likely to experience difficulty their first insertion attempt. Other variables associated with insertion difficulty included longer fingernails. Insertion refusal and difficulty affect use of the female condom for a sizable proportion of women. Women in this study who refused the self-insertion practice had greater aversion to inserting intravaginal barrier methods. Women who had initial difficulty inserting the female condom had a different profile from those who refused and can benefit from intensive skills training that includes supervised self-insertion practice. PMID- 11927119 TI - Barriers to use of oral contraceptives in ethnic Chinese women presenting for abortion. AB - The purpose of this study was to identify barriers to the use of oral contraceptives by ethnic Chinese women presenting for abortion. Using a qualitative descriptive study design, 40 ethnic Chinese women presenting for abortion were recruited. Data were collected in semi-structured interviews by one interviewer who is fluent in English, Mandarin, and Cantonese. Transcribed interviews were systematically analyzed to identify salient themes. The attitudes toward oral contraceptives were mostly negative. The most common fears were about weight gain, permanent infertility, and being considered "bad" (promiscuous). These mostly negative attitudes toward oral contraceptives appear to be barriers to their use. PMID- 11927120 TI - Challenging the stereotypes: men, withdrawal, and reproductive health in Lebanon. AB - In Lebanon, coitus interruptus or withdrawal remains a widely practiced method of family planning. Our research sought to understand the role of men in reproductive health in Lebanon by focusing on this common practice. Our main questions were: Why is it that the practice persists when more effective modern methods of family planning are available? How is the decision taken to practice withdrawal? When is withdrawal practiced and with whom? And, finally, does the practice of withdrawal affect sexual pleasure and the sexual relationship more generally?To answer these questions, we embarked on a small exploratory study using in-depth interviews with 16 open-ended questions. We found that the most important reason for the continuing practice of withdrawal is fear of side effects from other methods. Men and women expect pleasure and fulfillment in sexual relations, but they are willing to limit their pleasure to limit their fertility by means they consider safe. No one prototypical practice of withdrawal seems to exist, and this may explain whether or not the method fails to prevent pregnancy. PMID- 11927121 TI - Effect of Maytenus ilicifolia Mart. on pregnant mice. AB - Maytenus ilicifolia Mart. is used in Brazilian herbal medicine particularly for stomach disorders, but it is also used, as in other parts of South America, for fertility control. To verify its potential as an abortifacient, the lyophilized hydroalcoholic extract of its leaves was administered orally at a dose of 1000 mg/kg/day to mice between the first and third day of pregnancy (DOP), between the forth and sixth DOP, or between the seventh and ninth DOP. The extract caused a pre-implantation embryonic loss, but it did not have an effect on implantation or organogenesis. Morphological alterations of the reproductive system, not an embryotoxic effect, were not found. Estrogenic activity of the extract, exhibited by an uterotrophic effect, suggests that it may be interfering with the uterine receptivity to the embryo. PMID- 11927122 TI - The effect on MHC class II expression and apoptosis in placenta by IFNgamma administration. AB - To investigate the possible mechanisms by which interferon gamma (IFNgamma) affects pregnancy, the expression of class II major histocompatibility complex (MHC) antigens and cleavage of DNA that is a hallmark of apoptosis in the placenta were examined by molecular biochemical techniques, and progesterone levels were examined by radioimmunoassay. The semi-quantitative analysis with reverse transcription-polymerase chain reaction showed that the expression of MHC class II antigen in placenta increased when rabbits were treated with high doses of IFNgamma compared with the control. However, immunohistochemical study suggested that IFNgamma did not affect MHC class II expression in trophoblasts, but had a stimulatory effect on its expression in maternal decidua and placental lymphocytes. DNA fragmentation analysis and terminal deoxynucleotidyl mediated deoxyuridine triphosphate nick end labeling (TUNEL) assay indicated that the cleavage of DNA was detected in the placenta in both normal and IFNgamma-treated pregnancy. Quantitative analysis of apoptotic cells revealed an increase in trophoblasts treated with IFNgamma compared to those in normal pregnancy. Moreover, progesterone, which plays an important role in pregnancy, was reduced significantly in rabbits treated with IFNgamma. The results suggested that IFNgamma exerted its deleterious effect on pregnancy by inducing apoptosis in trophoblasts and by reducing the production of progesterone. PMID- 11927123 TI - Influence on coagulation activity by subcutaneous LMW heparin as an adjuvant treatment to fibrinolysis in acute myocardial infarction. AB - In this study, which includes 101 patients with acute ST segment-elevated myocardial infarction, we investigated the influence on the increased coagulation activity after streptokinase treatment by adding low-molecular-weight (LMW) heparin or placebo and the relation between the coagulation activity and ischemic episodes, coronary patency, and mortality. The expected increase of prothrombin fragment 1+2 (F1+2), thrombin-antithrombin (TAT), and D-dimer were significantly attenuated at 2, 6, and 18 h (D-dimer only at 18 h) in the dalteparin group compared to placebo. Ischemic episodes during the first 24 h appeared significantly more often in patients with F1+2 levels above the median at 18 h. There was a tendency to a lower frequency of Thrombolysis In Myocardial Infarction Trial (TIMI) grade 3 flow in the infarct-related artery in patients with TAT and D-dimer levels above the median at 18 h. F1+2, TAT, and D-dimer were significantly higher after 18, 6, and 18 h, respectively, in the deceased compared to surviving patients. Also, the lack of reduction of the levels of F1+2 between 6 and 18 h was related to a raised mortality. In conclusion, adjuvant treatment with LMW heparin to streptokinase attenuates increased coagulation activity. This might be of importance as remaining high coagulation activity is associated with signs of early reocclusion and raised mortality. PMID- 11927125 TI - Comparison of in vitro closure time (PFA-100) with whole blood electrical aggregometry and platelet surface antigen expression in healthy volunteers. AB - It was the aim of this study to compare in vitro closure time (PFA-100), reflecting platelet-related primary hemostasis, to more platelet-specific tests like whole blood electrical aggregometry and platelet surface antigen expression in healthy volunteers. In vitro closure time was measured using a PFA-100. Platelet surface antigen expression (CD63, CD62-P, CD42b, CD36, CD31) was determined in accordance with the consensus protocol for flow-cytometric characterisation of platelet function. Platelet aggregometry was performed using a whole blood electrical aggregometer (ADP and arachidonic acid as agonists). Analysis of the obtained data revealed only a few significant correlations between the different platelet function tests used. This finding can be explained by the various aspects of platelet function being focused by these tests in different extents. Whenever platelet function is analysed, the investigator should be aware of the specific and limited evidence of the method used. For screening purposes, it may be useful to introduce a platelet function index, referring to basal platelet activity, platelet adhesion and platelet aggregation at low and high shear stress forces. PMID- 11927124 TI - Antifactor Xa activity in intensive care patients receiving thromboembolic prophylaxis with standard doses of enoxaparin. AB - BACKGROUND: Low-molecular-weight heparins (LMWHs) have become increasingly used to prevent thromboembolic complications in intensive care patients. Unlike in medical and surgical patients, no data on the anticoagulant effectiveness of standard LMWH dosages exist in intensive care patients. Therefore, we prospectively investigated antifactor Xa (aFXa) levels after subcutaneous administration of 40 mg of enoxaparin in 89 intensive care patients over a 24-h period. METHODS: AFXa levels were measured before, 4, 12 and 24 h after subcutaneous administration of enoxaparin. Laboratory parameters including prothrombin time, activated partial thromboplastin time, antithrombin III, fibrinogen as well as platelet count were collected at same intervals. Demographics included age, sex, height, weight, body mass index, admission diagnosis, a thromboembolic risk score and a modified Goris multiple organ dysfunction score. RESULTS: At 4, 12 and 24 h, 56.5%, 39.3% and 12.6% of the study patients were within recommended antithrombotic aFXa levels (0.1-0.3 U ml( 1)). Presence of multiple organ dysfunction as well as high body weight were significantly correlated with low aFXa levels. CONCLUSION: European standard dosages of 40 mg of enoxaparin once daily proved to be ineffective in achieving recommended antithrombotic aFXa levels in intensive care patients. This was most pronounced in patients with high body weight and presence of multiple organ dysfunction. PMID- 11927126 TI - Clopidogrel-induced qualitative changes in thrombus formation correlate with stent patency in injured pig cervical arteries. AB - Thienopyridines (ticlopidine or clopidogrel) alone or in combination with aspirin are now the reference antiplatelet therapy after stent implantation. To better understand the high efficacy and low risk of bleeding with these agents, we tested clopidogrel alone or with aspirin in an acute ex vivo flow chamber model and in a subacute in vivo arterial thrombosis model. Clopidogrel induced a dose dependent increase in bleeding time (BT), inhibited ADP-induced platelet aggregation and in the flow chamber reduced thrombus size, and changed thrombus structure to broad-based structure composed of nondegranulated loosely attached platelets contrasting with the tight clumps of degranulated platelets seen without clopidogrel. The in vivo model involved angioplasty and stenting at the site of a preinduced arterial lesion and thrombosis in pig carotid arteries. Clopidogrel alone or with aspirin (but not aspirin alone) decreased the number of stented vessels occluded for more than 24 h and conversely reduced the number of occluding thrombus. At 96 h after stenting, 100% and 90% of the arteries were patent with clopidogrel/aspirin and clopidogrel alone, respectively (vs. 67% and 44% with aspirin and saline, respectively). Clopidogrel destabilizes thrombus without complete abolishment of platelet reactivity. PMID- 11927127 TI - Demonstration of inducible TFPI-2 mRNA synthesis in BeWo and JEG-3 trophoblast cells using a competitive RT-PCR. AB - Tissue factor pathway inhibitor-2 (TFPI-2) displays structural similarities with TFPI-1, the major inhibitor of tissue factor (TF)/, factor VIIa. It is synthesized mostly by syncytiotrophoblast in the placenta, but its physiological functions are not fully understood. We studied the synthesis of TFPI-2 mRNA and that of TFPI-1 and TF in three human trophoblast cell lines, JAR, BeWo, and JEG 3. We first developed specific competitive reverse transcription-polymerase chain reaction (RT-PCR) assays for each gene studied using human umbilical vein endothelial cells (HUVEC). The three trophoblast cell lines strongly synthesized TF mRNA whereas the synthesis of TFPI-1 mRNA was very low. TFPI-2 mRNA was not detected in unstimulated or stimulated JAR cells. In contrast, JEG-3 and, to a lesser extent, BeWo produced significant amounts of TFPI-2 mRNA, which were significantly increased after stimulation with phorbol 12-myristate 13-acetate (PMA). However, tumor necrosis factor-alpha (TNF-alpha) had no effect on this synthesis. JEG-3 and BeWo are thus two cell lines that could be used to study TFPI-2 gene regulation and to investigate the role of TF, TFPI-1, and TFPI-2 during trophoblast differentiation. PMID- 11927129 TI - Four missense mutations identified in the protein S gene of thrombosis patients with protein S deficiency: effects on secretion and anticoagulant activity of protein S. AB - Four missense mutations, G54R, T589I, K155E, and Y595C, were identified in the protein S (PS) gene of the patients with PS deficiency and venous thrombosis. Three patients were heterozygous for the novel mutations, G54R, T589I, and Y595C, while a remaining one patient was homozygous for the K155E mutation, which is known to be a polymorphism in the Japanese population. A family study revealed that the Y595C mutation was associated with a Type I PS deficiency and the K155E mutation with a Type II PS deficiency, while no family study was performed for the patients with the G54R and T589I mutations. To determine whether these four mutations play a causative role in PS deficiency, the four PS mutants and wild type PS were stably expressed in human embryo kidney (HEK) 293 cells. Pulse-chase experiments showed intracellular degradation and decreased secretion of the Y595C mutant. In the activated protein C (APC) cofactor assays, the specific activity of the K155E mutant decreased to 58% of that of wild-type PS. The APC cofactor activity of the three mutants, G54R, K155E, and T589I, were inhibited by C4b binding protein (C4BP) with a dose dependency similar to that of wild-type PS. These results indicate that the Y595C and the K155E mutations are responsible for a secretion defect and a decreased anticoagulant activity of PS, respectively. The remaining two mutations, G54R and T589I, however, did not produce any definite abnormality leading to a low plasma PS activity. PMID- 11927128 TI - Pharmacokinetics and pharmacodynamics of the prophylactic dose of enoxaparin once daily over 4 days in patients with renal impairment. AB - The pharmacokinetics of the low-molecular-weight heparin enoxaparin were evaluated in 12 healthy volunteers and 36 patients with mild, moderate or severe renal impairment. Enoxaparin was administered once daily by subcutaneous injections at a dose of 40 mg for 4 days and venous blood samples were taken over a 5-day period to determine antifactor Xa and antifactor IIa activity and the activated partial thromboplastin time. Adverse events were also recorded. The results for anti-Xa activity showed that the rate of absorption of enoxaparin was similar across the four groups of study participants. The elimination half-life increased with the degree of renal impairment, and this relationship was more evident after repeated dosing. Anti-Xa exposure was not significantly different between healthy volunteers and patients with mild or moderate renal impairment, but was significantly increased in patients with severe renal impairment (creatinine clearance < or =30 ml/min). Anti-Xa clearance decreased with the degree of renal impairment after repeated dosing, but only the difference between patients with severe renal impairment and healthy volunteers was statistically significant, the clearance on Day 4 being 39% lower in patients with severe renal impairment than in healthy volunteers (P=.0001). Anti-IIa activity was low in all study participants, and the activated partial thromboplastin time was not significantly different between the study groups. In conclusion, the clearance of enoxaparin is reduced in patients with severe renal impairment. Dose adjustment of enoxaparin may need to be recommended in these patients, but no recommendation can be made in patients with mild or moderate renal impairment. PMID- 11927130 TI - Markers of hemostatic system activation during treatment of deep vein thrombosis with subcutaneous unfractionated or low-molecular weight heparin. AB - Prothrombin fragments (F1+2), thrombin-antithrombin complexes (TAT) and D-dimers, markers of hemostatic system activation, were measured in 59 consecutive patients with deep vein thrombosis (DVT). Patients were randomly treated either with subcutaneous unfractionated heparin (UH) administered in two to three subcutaneous doses adjusted to activated partial thromboplastin time (APTT) or with low-molecular weight heparin (LMWH) (dalteparin) administered in a fixed dose of 200 IU/kg body weight in one subcutaneous injection daily. Before treatment, F1+2, TAT and D-dimer were above the cut-off level in 27/59 (46%), 34/59 (58%) and all (100%) patients, respectively. Significant associations were observed between F1+2 and TAT (r=.66, P<.001), TAT and D-dimer (r=.36, P<.005) and F1+2 and D-dimer (r=.30, P<.050). On the third day of treatment, F1+2 and TAT significantly decreased to reference values in almost all patients (in 64/66 determinations of both F1+2 and TAT) and remained low on the seventh day of treatment. Compared to pretreatment values, a nonsignificant decrease of D-dimer was noted in both groups, but all values remained above the cut-off value. When markers of hemostatic system activation in the UH and LMWH groups were compared, no significant differences were observed. It was concluded that subcutaneous UH in an APTT-adjusted dose and subcutaneous LMWH in a once-daily weight-adjusted dose controlled these markers of hemostatic system activation in a similar manner. PMID- 11927131 TI - Differences between neonates and adults in carbohydrate sequences and reaction kinetics of plasmin and alpha(2)-antiplasmin. AB - This study investigates reaction kinetics by slow-binding kinetics methods of both adult and fetal plasmin (Types 1 and 2) with adult and fetal alpha(2) antiplasmin. In addition, carbohydrate sequences of Fetal and Adult Plasminogen Types 1 and 2, as well as fetal and adult alpha(2)-antiplasmin, were determined by mass spectrometric analysis. All curves of plasmin-alpha(2)-antiplasmin interaction followed the same pattern, indicating reversible slow-binding inhibition with an initial loose complex and a following tight complex. Differences between fetal and adult plasmin reactions with alpha(2)-antiplasmin were predominantly due to the initial loose complex. Values for K(i initial) in the reaction with adult alpha(2)-antiplasmin were 1.5 and 1.6 nM for Fetal Plasmin Types 1 and 2, respectively; compared to 0.3 and 0.7 nM for the corresponding adult types. Increasing concentrations of tranexamic acid resulted in a continuous increase of K(i initial) until a plateau was reached which was similar for all plasmin types. Almost identical values could be obtained when fetal alpha(2)-antiplasmin was used instead of adult alpha(2)-antiplasmin. Mass spectrometric analyses of the glycans present on plasminogen revealed a higher level of truncated N-glycans on the fetal material compared to the adult. The O glycans of fetal and adult plasminogen were closely similar and only minor differences were observed between N-glycans of fetal and adult alpha(2) antiplasmin. In conclusion, both fetal plasmin isoforms are less inhibited by alpha(2)-antiplasmin compared to the adult plasmin variants. These findings are important for the understanding of the physiology of the fibrinolytic system in neonates and provide further evidence that differences in glycosylation could be associated with marked effects on protein function. PMID- 11927132 TI - Plasma and serum levels of D-dimer and their correlations with other hemostatic parameters in pregnancy. AB - The measurement of D-dimer in serum samples (s-DD) after standardized coagulation has been reported as a possible single global test for fibrinolysis in unstimulated conditions in healthy subjects and in patients with ischemic heart disease and with different metabolic disorders. No study has been performed on the use of this test in pregnancy, a condition characterized by physiological changes both in coagulation and in fibrinolysis. In this preliminary study, we have evaluated in 28 women with physiological pregnancy and in 23 comparable controls s-DD and a number of markers of coagulation and fibrinolysis. In nonpregnant women s-DD showed a good correlation with fibrinolytic parameters [euglobulin lysis time (ELT) and type 1 inhibitor of tissue plasminogen activator (PAI-1) act: P<.01; tissue plasminogen activator (t-PA) ag and PAI-1 ag: P<.05], confirming previous data, whereas in pregnant women no correlation was observed. Plasma DD (pls-DD) and s-DD levels were not correlated either in pregnant or in control women. s-DD levels were significantly higher than pls-DD in controls and in 15/28 pregnant women whose pls-DD values were in normal range or mildly increased (<110 ng/ml; P<.05), whereas in the 13 pregnant women with high pls-DD levels no significant differences were found between pls-DD and s-DD levels. Because in pregnancy high pls-DD levels are frequently found, possibly only as a consequence of enhanced clotting activation and fibrin deposition, we cannot exclude that D-dimer measured in serum reflects, at least in part, cross-linked fibrin degradation products (FDP) already present in blood before standardized coagulation. Therefore, D-dimer generated in vitro would account only in part for s-DD measured. This can explain why in pregnant women, differently from controls, s-DD does not correlate with fibrinolytic parameters. In conclusion, this preliminary study indicates that baseline pls-DD levels may be an important potential confounder in the interpretation of s-DD results in pregnant women and that s-DD cannot be proposed as a tool for a rapid evaluation of fibrinolytic activity in pregnancy. PMID- 11927133 TI - Effect of a synthetic carboxy-terminal peptide of alpha(2)-antiplasmin on urokinase-induced fibrinolysis. AB - alpha(2)-Antiplasmin (alpha(2)AP) interferes with the binding of plasminogen to fibrin because lysine residues in its carboxy-terminal region compete with those in fibrin, presumably the same way that free lysine or epsilon-aminocaproic acid (EACA) inhibits plasminogen binding to fibrin. While this overall process causes an inhibition of fibrinolysis, the converse was observed with a 26-residue synthetic peptide (AP26) corresponding to the carboxy-terminal region of alpha(2)AP. The AP26 peptide, in fact, accelerated urokinase-induced lysis of (1) fully crosslinked fibrin with complete gamma-dimer and alpha-polymer formation; (2) partially crosslinked fibrin that had undergone only gamma-dimerization; and (3) noncrosslinked fibrin. The AP26 peptide also inhibited factor XIIIa-catalyzed crosslinking of fibrin alpha-chains, and this also accelerated lysis of fibrin. EACA had no effect. In the presence of noncrosslinked fibrin, AP26 promoted plasminogen activation by urokinase and fibrinolysis. EACA only slightly increased the rate of plasminogen activation, and as expected, it inhibited fibrinolysis. Since AP26 peptide enhanced the lysis of partially crosslinked and noncrosslinked fibrin, our results indicate that inhibition of factor XIIIa catalyzed alpha-polymer formation by AP26, although associated with accelerated fibrinolysis, is not the primary mechanism. Instead, our data support the conclusion that AP26 enhances the conversion of plasminogen to plasmin approximately 5-fold, probably by inducing a conformational change in plasminogen structure just as occurs with low concentrations of lysine or EACA. At higher concentrations, however, AP26 apparently does not approach the avidity or affinity of lysine or EACA for the kringle structures of plasminogen or plasmin so that their binding to fibrin is blocked. Whether AP26 alone, or as part of another molecule, could have potential for enhancing thrombolysis will require further study. PMID- 11927134 TI - Suitability of four polymorphic DNA markers for indirect genetic diagnosis of haemophilia A in Japanese subject. AB - Indirect genetic diagnosis using polymorphic DNA markers can detect carriers of haemophilia A (HA). These markers include CA repeat polymorphisms at intron 13 (CA-13) and CT-AG at intron 22 of the coagulation factor VIII (FVIII) gene, and also certain restriction fragment length polymorphisms (RFLPs) such as HindIII at intron 19 and BclI at intron 18. Recently, CA repeat polymorphism at intron 6 (CA 6) was also reported. We compared usefulness of the BclI RFLP, the HindIII RFLP, and CA-13 to that of CA-6. Heterozygosity of markers was examined in 282 X chromosomes obtained from 205 subjects including HA patients. Expected heterozygosity of the HindIII and BclI RFLPs was 30.0% and 27.9%, while observed heterozygosity was 28.0% and 26.5%. Expected heterozygosity of CA-13 was 45.6%, and observed heterozygosity was 40.0%. CA-6 showed two alleles with repeat numbers of 13 and 14, expected and observed heterozygosity were low (1.4% and 2.6%, respectively). When we used these markers in a HA lineage where the mother was a carrier according to coagulation factor assays, carrier diagnosis was possible using CA-13, the HindIII RFLP, and the BclI RFLP. This was not true for CA-6, for which the mother was homozygous. Although CA-13, and the HindIII and BclI RFLPs were useful for indirect genetic diagnosis of HA, CA-6 proved less useful because of low heterozygosity. PMID- 11927135 TI - Analysis of monoamine oxidase A (MAOA) promoter polymorphism in Finnish male alcoholics. AB - Alterations in monoamine oxidase A (MAOA) expression and enzyme activity may be associated with alcoholism and impulsive behavior. Therefore, functional polymorphisms in the MAOA gene would be good candidates to consider in the interindividual differences that exist in the susceptibility to alcoholism. One variant that has been considered as a candidate in alcoholism is a repeat polymorphism in the MAOA gene promoter. We analyzed a cohort of Finnish males with either type 1 or type 2 alcoholism, as well as controls, for differences in the distribution of MAOA promoter alleles. Based on other studies, we postulated that type 2 alcoholism, which is associated with antisocial behavior, but not type 1 alcoholism, would be correlated with the inheritance of the low promoter activity allele. However, we failed to find a difference in allele distribution in type 1 and type 2 alcoholics. In addition, there was no difference in the allele distribution when each group of alcoholics was compared with controls. However, when both groups of alcoholics were pooled and compared with controls, the difference in allele distribution reached a trend towards significance. Our results suggest a minimal association between the MAOA low activity promoter alleles and alcoholism, regardless of the presence or absence of antisocial behavior. Interestingly, approximately 3% of type 2 alcoholics were found to be heterozygous for the MAOA promoter polymorphism. Since MAOA is X-linked, the heterozygotes are probable cases of Klinefelter's syndrome (47,XXY) suggesting that X-chromosome aneuploidy may increase the risk for developing type 2 alcoholism. PMID- 11927136 TI - Mutation screening of the human Clock gene in circadian rhythm sleep disorders. AB - We tested whether the human Clock (hClock) gene, one of the essential components of the circadian oscillator, is implicated in the vulnerability to delayed sleep phase syndrome (DSPS) and non-24-hour sleep-wake syndrome (N-24). Screening in the entire coding region of the hClock gene with PCR amplification revealed three polymorphisms, of which two predicted the amino acid substitutions R533Q and H542R. The frequencies of the R533Q and H542R alleles in patients with DSPS or N 24 were very low and not significantly different from those in control subjects. A T3111C polymorphism in the 3'-untranslated region of hClock, which had been reportedly associated with morning or evening preference for activity, was also investigated; the results showed that the 3111C allele frequency decreased in DSPS. Polymorphisms in the coding region of the hClock gene are unlikely to play an important role in the development of DSPS or N-24. The possible contribution of the T3111C polymorphism to DSPS susceptibility should be studied further. PMID- 11927137 TI - The tryptophan depletion test: impact on sleep in primary insomnia - a pilot study. AB - The application of the tryptophan depletion test is based on the assumption that the decrease of plasma or serum tryptophan concentration following the ingestion of a tryptophan-free amino acid drink reflects a central nervous effect on serotonin metabolism. In the present study the impact of tryptophan depletion on polysomnographically recorded sleep in patients with primary insomnia was studied. Fifteen patients with primary insomnia slept for four nights in the sleep laboratory. Prior to the fourth night the tryptophan depletion test was applied. Sleep EEG variables served as outcome parameters. Patients with primary insomnia, compared to baseline values showed a highly significant decrease of serum tryptophan concentrations after the amino acid drink. Concerning sleep parameters, stage 1 (% sleep period time=SPT) was increased, whereas stage 2 (% SPT) was decreased. Indices of phasic activity of rapid eye movement (REM) sleep (REM density) were increased after the tryptophan depletion compared to baseline. The results suggest a negative impact of tryptophan depletion on sleep continuity and a stimulating effect on phasic measures of REM sleep in patients with primary insomnia. PMID- 11927138 TI - Effect of risperidone on sleep in schizophrenia: a comparison with haloperidol. AB - The aim of the present study is to determine the effect of the atypical antipsychotic drug, risperidone on sleep measures in patients with schizophrenia by polysomnography. Sleep measures were compared in five schizophrenic patients who were receiving risperidone alone and five schizophrenic patients who were receiving haloperidol alone. There were no differences between these two groups in their demographic characteristics or doses of antipsychotic medication. The slow wave sleep period was significantly longer in the risperidone-treated group than in the haloperidol-treated group. There were, however, no other significant differences in sleep variables between these groups. This difference in the effect on sleep between risperidone and haloperidol may be due to their differential actions on serotonin (5-HT2) receptors. Risperidone, which is known to be a serotonin-dopamine antagonist, has the potential to improve the quality of sleep in schizophrenic patients. PMID- 11927139 TI - Decreased serum brain-derived neurotrophic factor levels in major depressed patients. AB - Recent findings with animal models have suggested a possible role for brain derived neurotrophic factor (BDNF) in depression. We have therefore hypothesized that depression could be characterized by low levels of serum BDNF. Major depressed patients (15F + 15M) diagnosed according to DSM-IV criteria and healthy controls (15F + 15M) participated in the study. Serum BDNF was assayed with the ELISA method and the severity of depression was evaluated with Montgomery-Asberg Depression Rating Scale (MADRS). BDNF levels were significantly lower in patients than in controls: 22.6 +/- 3 and 26.5 +/- 7 ng/ml (t-test = 2.7; d.f. = 58; P < 0.01). They were negatively correlated to the MADRS scores (r = -0.55; P < 0.02). Female patients were more depressed and released less BDNF than men. Analysis of covariance (MADRS and gender as independent variable vs. BDNF as dependent variable) indicated that depression severity mainly accounted for the negative correlation. These results suggest that major depression is characterized by low serum BDNF levels and support the hypothesis of neurotrophic factor involvement in affective disorders. PMID- 11927140 TI - Social perception in schizophrenia: the role of context. AB - The purpose of this study was to examine social context processing in persons with schizophrenia. A secondary goal was to examine the ecological validity of these measures (i.e. how they relate to social behavior in the treatment setting). The performance of 35 persons with DSM-IV diagnosed schizophrenia and/or schizoaffective disorder was compared to a non-clinical control sample of 46 individuals on a battery of social perceptual tasks that require social context processing. In addition, the relationship between social context processing and ward behavior (as measured with the Nurse's Observation Scale for Inpatient Evaluation) was examined for the clinical sample only. The results showed that the group with schizophrenia/schizoaffective disorder was impaired on all tasks relative to the control group and showed little evidence of utilizing available contextual information. Task performance for the group with schizophrenia/schizoaffective disorder was relatively independent of symptoms, but was related to social functioning in the treatment setting. Implications for future research are discussed. PMID- 11927141 TI - Age, expressed emotion, and interpersonal control patterning in families of persons with schizophrenia. AB - Expressed emotion (EE) is a measure of a relative's attitude toward a person with a mental disorder as reflected by comments made to an interviewer. Over the years, an impressive body of research has been generated in attempts to explain the relationship of EE and course of illness, particularly in regard to schizophrenia. Past analyses have demonstrated two common patterns of interpersonal control in families with high-EE relatives. The first was that the relative and patient competed for 'who's in charge', with both rigidly asserting control. The other was that the relatives responded rigidly in one control mode no matter what the offspring was saying. In this study, we tested whether the difference in patterning was due to the patients being older. Study participants were 71 relative-patient dyads. EE was measured with the Five-Minute Speech Sample, the Family Problem-Solving Task was used to generate interaction, and interaction data were coded with the Relational Control Coding System. Multilevel modeling was used to estimate a model with verbal exchanges as Level 1, relatives' EE as Level 2, and age of the patient as Level 3. Results showed that age attenuated the relationship between the patient's message and the parent's response, and this effect was stronger in families with high-EE relatives. This effect may reflect disengagement on the part of the relative, the patient, or both. The over-responsive families may need advice on how to create more distance, or how to be more objective, but the distant families may need more support and encouragement to communicate and problem-solve. PMID- 11927142 TI - Event-related potential dysfunction in posttraumatic stress disorder: the role of numbing. AB - The purpose of this study was to examine the relationship between disturbance in event-related potentials (ERPs) and symptom clusters in posttraumatic stress disorder (PTSD). ERPs were recorded in 17 unmedicated civilian PTSD patients and 17 age- and sex-matched controls during a conventional auditory oddball task. PTSD symptom clusters (re-experiencing, active avoidance, numbing, hyperarousal) were correlated with ERP measures. The PTSD group showed ERP disturbances to target stimuli (reduced P200 and P300 and increased N200 amplitude, increased N200 and P300 latency) and reduced P200 amplitude to common stimuli compared to the control group. A significant negative correlation was found between the intensity of numbing symptoms and parietal P300 amplitude. This study replicates findings of disturbed N200 and P300 components in PTSD, reflecting impairments in stimulus discrimination and attention. The finding that numbing was associated with reduced attention processing (P300) is consistent with models positing a relationship between disordered arousal and attention in PTSD. PMID- 11927143 TI - Drug use disorders in Japanese eating disorder patients. AB - A previous questionnaire study suggested that drug use disorder (DUD: abuse/dependence on drugs, other than alcohol) in Japanese eating disorder (ED) patients was less prevalent than in Western countries, although eating and drug use disorders have spread simultaneously in Western countries. However, the precise prevalence and comorbidity features remain unknown. Subjects consisted of 62 patients with anorexia nervosa restricting type; 48 patients with anorexia nervosa binge eating/purging type; and 75 patients with bulimia nervosa purging type. The Japanese version of the Structured Clinical Interview for DSM-III-R; the Structured Clinical Interview for DSM-III-R Personality Disorders; and the supplement module of the Schedule for Affective Disorders and Schizophrenia Lifetime version were used for the interview. Sixteen (8.6%, 95% CI = 4.6-12.7%) patients had lifetime diagnoses of DUD. Drugs were solvent fumes or benzodiazepines, and only one patient had been dependent on methamphetamine. More than half of the patients with lifetime DUD diagnoses were multi-impulsivitists. On multivariate analysis, DUD was significantly linked with childhood parental loss, history of conduct disorder and borderline personality disorder. Thus, the prevalence of DUD in Japanese ED patients was indeed lower than that in Western countries. However, similar comorbidity was found in ED patients with DUD compared with that of those in Western countries. The current study suggests that ED and DUD have different origins, although they share the feature of impulsivity. Further study in the general population is needed to clarify these issues. PMID- 11927144 TI - Dimensional structure of the Yale-Brown Obsessive-Compulsive Scale (Y-BOCS). AB - The Yale-Brown Obsessive-Compulsive Scale (Y-BOCS) is a widely used instrument to assess obsessive-compulsive symptomatology. The present study provides evidence that the Y-BOCS is best represented by a three-dimensional model comprising severity of obsessions (factor 1), severity of compulsions (factor 2) and resistance to symptoms (factor 3). On the basis of exploratory factor analysis, this structure was found for both baseline (n = 109) and discharge ratings (n = 68) following a multimodal cognitive-behavioral intervention. The factor solution remained essentially unchanged when two optional items (items 1b and 6b) were dropped from analysis. The three-factor structure was replicated with confirmatory factor analysis and showed better fit than previously proposed single- and two-factor models. For future research, we propose a new Y-BOCS scoring algorithm that takes this factor structure into account. A further result was that resistance significantly declined in response to cognitive-behavioral intervention, whereas drug treatment alone did not seem to moderate this variable according to previous research conducted by Kim et al. [Psychiatry Research 51 (1994) 203-211]. PMID- 11927145 TI - Prolactin but not ACTH increases during sodium lactate-induced panic attacks. AB - Paradoxically, the pituitary-adrenal axis is not activated during sodium lactate induced panic. We measured the response of another stress-sensitive hormone, prolactin, to standard lactate and placebo infusion in a double-blind randomised design in eight patients with panic disorder and eight matched normal controls. Prolactin release was significantly elevated (P < 0.05) in panickers compared with non-panickers, whereas ACTH secretion was not activated at all. This differential stress response needs further investigation. PMID- 11927146 TI - Beta-thromboglobulin and platelet factor 4 levels in post-myocardial infarction patients with major depression. AB - Platelet factor 4 (PF 4) and beta-thromboglobulin (beta-TG) were studied in 12 depressed post-myocardial infarction (MI) patients and 12 matched non-depressed post-MI patients. PF4 was significantly higher in the depressed group than in the non-depressed group. beta-TG was increased in the depressed subgroup, but the difference was not statistically significant. PMID- 11927147 TI - Expression of netrin-1 and netrin-1 receptor, DCC, in the rat olfactory nerve pathway during development and axonal regeneration. AB - Netrin-1 is a bifunctional secreted protein that directs axon extension in various groups of developing axonal tracts. The transmembrane DCC (deleted in colorectal cancer) receptor is described as netrin-1 receptor and is involved in the attractive effects of netrin-1. In this study, we examined the spatio temporal expression patterns of both netrin-1 and DCC in the rat olfactory system at different stages of development and during axonal regeneration following unilateral bulbectomy. High DCC expression was detected on the pioneer olfactory axons as they are extending toward the telencephalon. This expression was transient since from embryonic day 16 onwards, DCC was no longer detected along the olfactory nerve path. From embryonic day 14 until birth, DCC was also expressed within the mesenchyme surrounding the olfactory epithelium. During the same period, netrin-1 protein was detected along the trajectory of olfactory axons up to the olfactory bulb and its expression pattern in the nasal mesenchyme largely overlapped that of DCC. Moreover, netrin-1 continued to be present during the two first post-natal weeks, and a weak protein expression still persisted in the dorso-medial region of the olfactory epithelium in adult rats. While unilateral bulbectomy induced a transient up-regulation of netrin-1 in the lamina propria, particularly in the dorso-medial region of the neuroepithelium, no DCC expression was detected on the regenerating olfactory axons. In the developing olfactory bulb, the extension of mitral cell axons was associated with DCC presence while netrin-1 was absent along this axonal path. DCC was also highly expressed in the newly formed glomeruli after birth, and a weak DCC expression was still detected in the glomerular layer in adult rats. Taken together, these data support the notion that netrin-1, via DCC expressed on axons, may play a role in promoting outgrowth and/or guidance of pioneering olfactory axons toward the olfactory bulb primordium. Moreover, association of netrin-1 with mesenchymal DCC may provide a permissive environment to the growth of both pioneer and later growing axons. The maintenance of netrin-1 expression in the nasal mesenchyme of adult rats as well as its regional up-regulation following unilateral bulbectomy infer that netrin-1, even in the absence of DCC, may be involved in the process of axonal growth of newly differentiated olfactory receptor neurons probably through the use of other receptors. PMID- 11927148 TI - Neural inhibition by c-Jun as a synergizing factor in bone morphogenetic protein 4 signaling. AB - The transcription factor, activator protein 1 (AP-1) complexes (c-Jun and c-Fos heterodimers) has been shown to interact with transforming growth factor beta signaling in mammalian cells and Drosophila embryo. Here we show that c-Jun alone is involved in the anti-neuralizing activity of bone morphogenetic protein 4, a transforming growth factor beta superfamily member, in Xenopus neurogenesis. Co injection of mRNAs encoding c-jun and a dominant negative bone morphogenetic protein receptor completely inhibits dominant negative bone morphogenetic protein receptor-induced neuralization and reverses the epidermal fate in the animal cap. Surprisingly, a dominant negative c-Jun does not induce neural tissue in the animal cap, but it synergizes with dominant negative bone morphogenetic protein receptor for neural induction. Temporal analysis using a dexamethasone-inducible c-Jun shows that exogenous c-Jun activity must be turned on before or at stage 11 to fulfill the anti-neuralizing effect. Neural inhibition by c-Jun does not occur until stage 13 suggesting that c-Jun probably acts by suppressing neural maintenance rather than neural initiation. This is also supported by the fact that c-Jun does not inhibit expression of the neural-initializing gene Zic-r1 but the neural cofactor Sox2, and that ectopic expression of Sox2 attenuates the anti neuralizing effect of c-Jun. Finally, we display that the c-Jun effect is enhanced by an auto-regulatory loop between c-Jun and bone morphogenetic protein. These studies suggest that c-Jun/AP-1 is a converging point in both the fibroblast growth factor and transforming growth factor beta signaling pathways. Based on our findings, we propose that c-Jun synergizes with bone morphogenetic protein 4 signaling to inhibit neural development in Xenopus ectoderm. PMID- 11927149 TI - A distinct form of calcium release down-regulates membrane excitability in neocortical pyramidal cells. AB - We reported a novel type of calcium release from inositol-1,4,5-trisphosphate (IP(3))-sensitive calcium stores synergistically induced by muscarinic acetylcholine receptor (mAchR)-mediated increase in IP(3) and action potential induced calcium influx (IP(3)-assisted calcium-induced calcium release, IP(3) assisted CICR). To clarify its functional significance, the effects of IP(3) assisted CICR on spike-frequency adaptation were examined in layer II/III neurons from rat visual cortex slices. IP(3)-assisted CICR was enabled with a high concentration of the mAchR agonist carbachol (10 microM). The magnitude of this CICR was the more augmented at higher firing frequencies. With 10 microM carbachol, spike-frequency adaptation was reduced for spike trains at 'low' firing frequencies (6-10 Hz), but was rather enhanced at 'high' firing rates (16 22 Hz): excitability was down-regulated at 'high' frequencies. With 1 microM carbachol, by contrast, IP(3)-assisted CICR failed to occur, and spike-frequency adaptation was always reduced at any spike frequencies. Intracellular injection of the IP(3) receptor blocker heparin prevented both the mAchR-mediated occurrence of IP(3)-assisted CICR and enhancement of spike-frequency adaptation with 10 microM carbachol. Both of these mAchR-mediated effects were reproduced by intracellular IP(3) injection, and were shown to be associated with each other by simultaneous recordings of membrane potential and intracellular calcium increase. We propose that IP(3)-assisted CICR offers a novel way to protect these cortical neurons from hyperexcitability and presumably from excitotoxic cell death. PMID- 11927150 TI - Emergence of a functional coupling between inositol-1,4,5-trisphosphate receptors and calcium channels in developing neocortical neurons. AB - Cortical pyramidal neurons are considered to be less excitable in the immature cortex than in adults. Our previous report revealed that a negative feedback regulation of membrane excitability is highly correlated with a novel form of calcium release from inositol-1,4,5-trisphosphate (IP(3))-sensitive calcium stores (IP(3)-assisted calcium-induced calcium release) in neocortical pyramidal neurons under muscarinic cholinergic activation. As a step to understand the ground for the low membrane excitability in immature tissue, we examined development of IP(3)-assisted calcium-induced calcium release. In visual cortex neurons from 'juvenile' rats (2-3 weeks of age), an enhancement of spike frequency adaptation occurred at high spike-frequencies (16-22 Hz), whereas the reduction was observed at low frequencies (6-10 Hz). IP(3)-assisted calcium induced calcium release occurred at the higher frequencies only. In 'early' postnatal tissue (1 week of age), by contrast, at neither high nor low frequencies did this form of calcium release occur, and muscarinic cholinergic activation always induced a reduction of spike-frequency adaptation at any spike frequencies. The mechanism for the failure of induction of IP(3)-assisted calcium induced calcium release in 'early' postnatal tissue was investigated. Both an ample supply of calcium influx, elicited by higher frequency spike trains, and a supplementary injection of IP(3) through whole-cell pipets, combined together or applied alone, failed to enable IP(3)-assisted calcium-induced calcium release in 'early' postnatal tissue. Muscarinic cholinergic activation alone induced a conventional IP(3)-induced calcium release similar to that observed in neurons from 'juvenile' tissue. Together, it is most likely that functional IP(3)Rs and calcium channels are already present and functional, but are not yet adequately assembled to allow IP(3)-assisted calcium-induced calcium release in cortical pyramidal neurons from rats of 1 week old. PMID- 11927151 TI - Excitatory and inhibitory neurons express c-Fos in barrel-related columns after exploration of a novel environment. AB - Recent work has shown that behaviorally meaningful sensory information processing is accompanied by the induction of several transcription factors in the barrel cortex of rodents. It is now generally accepted that stimulus-transcription coupling is an important step in the sequence of events leading to long-term plastic changes in neuronal structure and function. Nevertheless, so far few data are available as to what types of neurons are involved in such a genomic response. Here, we determined the morphological and neurochemical identity of neurons in rat barrel cortex showing a c-Fos-immunoreactive nucleus after exploration of an enriched environment. Double stainings of c-Fos and glial fibrillary acidic protein excluded astrocytes as a possible cell type expressing this transcription factor. By morphological phenotyping with intracellular Lucifer Yellow injections, it was found that a large majority were probably excitatory pyramidal cells, but inhibitory interneurons were also found to contain c-Fos-immunoreactive nuclei. By neurochemical phenotyping of GABAergic interneurons with specific antibodies, a significant induction was found, in a layer-dependent manner, for the populations of glutamic acid decarboxylase-, parvalbumin-, calbindin- and vasoactive intestinal polypeptide-immunoreactive neurons but not for calretinin-immunoreactive cells in experimental compared to control columns. From these data we conclude that thalamic afferents effectively drive cortical excitatory as well as inhibitory intracortical circuits. Thus, the adaptations of receptive field properties of cortical neurons after different manipulations of the sensory periphery are likely to be caused by plastic changes in excitatory and inhibitory networks. PMID- 11927152 TI - Perceptual changes in illusory wrist flexion angles resulting from motor imagery of the same wrist movements. AB - Recent neuroimaging studies have suggested that similar cortical motor areas are recruited both by kinesthetic sensations elicited by tendon vibration and by voluntarily imaging one's own movements of the same joints. Little is known, however, as to whether kinesthetic motor imagery interacts with kinesthetic illusion. We examined such interaction by behavioral analysis in which 19 subjects imagined wrist flexion or extension, with or without illusory flexion induced by tendon vibration. Electromyograms were also recorded to monitor the peripheral modulations caused by the interaction. The kinesthetic motor imagery had a psychophysical effect on kinesthetic illusion in the absence of overt movement. It was confirmed that the subjects could imagine wrist movements without facilitating muscle activities in the absence of vibration stimuli. The electromyogram activity of the vibrated extensor muscles was significantly higher than that of non-vibrated flexor muscles. Motor imagery of wrist extension, when illusory flexion was experienced, reduced the angle of illusory flexion while enhancing extensor muscle activities in comparison with the control. On the other hand, flexion motor imagery increased the angle of illusory flexion with or without enhancement of flexor muscle activities. Our results indicate that motor imagery interacts with kinesthetic illusion with or without enhancement of activities of the related muscles. This suggests (1) that common neural substrates shared by imagery and by illusion exist and (2) that different physiological mechanisms contribute to the enhancement of muscle activities of vibrated muscles and their antagonists. PMID- 11927153 TI - Decreased [(3)H]spiperone binding in the anterior cingulate cortex of schizophrenia patients: an autoradiographic study. AB - Abnormalities in the anterior cingulate cortex have been reported in patients with schizophrenia, and have been implicated in the pathophysiology of this disorder. In the present study, we have examined antipsychotic-sensitive binding sites in the left anterior cingulate cortex of schizophrenia patients and controls. Using quantitative autoradiography and [(3)H]spiperone as a ligand, both saturation and competition experiments were performed in post-mortem brain tissue obtained from six schizophrenia and six control cases. Saturation experiments revealed that the maximum number of [(3)H]spiperone binding sites was significantly reduced by 31% in the schizophrenia group as compared to the control group (65.3+/-5.6 fmol/mg tissue versus 94.2+/-7.3 fmol/mg tissue). Increased dissociation constant was also observed in the schizophrenia group (2.2+/-0.4 nM versus 1.3+/-0.2 nM), but was not statistically significant (P=0.07). Competition experiments were performed in order to examine the pharmacological profile of [(3)H]spiperone binding, and revealed that: (i) displacement of [(3)H]spiperone binding by clozapine and mianserin was significantly reduced in the schizophrenia group as compared to the control group (-26% and -16% respectively); (ii) the order of displacement potency of the drugs tested was: haloperidol>mianserin>butaclamol approximately risperidone>clozapine>2-amino-6,7-dihydroxy-1,2,3,4-tetrahydronaphthalene. Our results suggest a reduction of antipsychotic-sensitive binding sites in the anterior cingulate cortex of patients with schizophrenia. Such abnormality could lead to an imbalance in neurotransmitter regulation in the anterior cingulate cortex which may contribute to the emergence of some symptoms of schizophrenia. PMID- 11927154 TI - Adrenomedullin expression is up-regulated by ischemia-reperfusion in the cerebral cortex of the adult rat. AB - Changes in the pattern of adrenomedullin expression in the rat cerebral cortex after ischemia-reperfusion were studied by light and electron microscopic immunohistochemistry using a specific antibody against human adrenomedullin (22 52). Animals were subjected to 30 min of oxygen and glucose deprivation in a perfusion model simulating global cerebral ischemia, and the cerebral cortex was studied after 0, 2, 4, 6, 8, 10 or 12 h of reperfusion. Adrenomedullin immunoreactivity was elevated in certain neuronal structures after 6-12 h of reperfusion as compared with controls. Under these conditions, numerous large pyramidal neurons and some small neurons were intensely stained in all cortical layers. The number of immunoreactive pre- and post-synaptic structures increased with the reperfusion time. Neurons immunoreactive for adrenomedullin presented a normal morphology whereas non-immunoreactive neurons were clearly damaged, suggesting a potential cell-specific protective role for adrenomedullin. The number and intensity of immunoreactive endothelial cells were also progressively elevated as the reperfusion time increased. In addition, the perivascular processes of glial cells and/or pericytes followed a similar pattern, suggesting that adrenomedullin may act as a vasodilator in the cerebrocortical circulation. In summary, adrenomedullin expression is elevated after the ischemic insult and seems to be part of CNS response mechanism to hypoxic injury. PMID- 11927155 TI - Altered receptor subtypes in the forebrain of GABA(A) receptor delta subunit deficient mice: recruitment of gamma 2 subunits. AB - A GABA(A) receptor delta subunit-deficient mouse line was created by homologous recombination in embryonic stem cells to investigate the role of the subunit in the brain GABA(A) receptors. High-affinity [(3)H]muscimol binding to GABA sites as studied by ligand autoradiography was reduced in various brain regions of delta(-/-) animals. [(3)H]Ro 15-4513 binding to benzodiazepine sites was increased in delta(-/-) animals, partly due to an increment of diazepam insensitive receptors, indicating an augmented forebrain assembly of gamma 2 subunits with alpha 4 subunits. In the western blots of forebrain membranes of delta(-/-) animals, the level of gamma 2 subunit was increased and that of alpha 4 decreased, while the level of alpha1 subunits remained unchanged. In the delta( /-) forebrains, the remaining alpha 4 subunits were associated more often with gamma 2 subunits, since there was an increase in the alpha 4 subunit level immunoprecipitated by the gamma 2 subunit antibody. The pharmacological properties of t-butylbicyclophosphoro[(35)S]thionate binding to the integral ion channel sites were slightly altered in the forebrain and cerebellum, consistent with elevated levels of alpha 4 gamma 2 and alpha 6 gamma 2 subunit-containing receptors, respectively.The altered pharmacology of forebrain GABA(A) receptors and the decrease of the alpha 4 subunit level in delta subunit-deficient mice suggest that the delta subunit preferentially assembles with the alpha 4 subunit. The delta subunit seems to interfere with the co-assembly of alpha 4 and gamma 2 subunits and, therefore, in its absence, the gamma 2 subunit is recruited into a larger population of alpha 4 subunit-containing functional receptors. These results support the idea of subunit competition during the assembly of native GABA(A) receptors. PMID- 11927156 TI - A 72 kDa heat shock protein is protective against the selective vulnerability of CA1 neurons and is essential for the tolerance exhibited by CA3 neurons in the hippocampus. AB - The correlation between the expression of a 72 kDa heat shock protein and vulnerability of hippocampal CA1, CA3, and dentate gyrus regions to glutamate toxicity was investigated using a highly specific antisense oligonucleotide technique. Glutamate (1 mM, 15 min) caused region-dependent neuronal damage in cultured hippocampal slices 24 h after exposure and the most severe damage was observed in CA1. When slices were heat-shocked (43.5 degrees C, 30 min) before exposure to glutamate, neuronal damage in CA1 was attenuated. The strongest protection was observed when the interval between the heat shock and the exposure to glutamate was 3 days, which coincided with the maximal induction of a 72 kDa heat shock protein in neurons. When the expression of a 72 kDa heat shock protein was suppressed by the antisense oligonucleotide, the protective effect of the heat shock was completely inhibited. Glutamate itself also induced a 72 kDa heat shock protein in neurons, region-dependently, 24 h after the exposure. The signal of a 72 kDa heat shock protein in CA3 and dentate gyrus was significantly stronger than that in CA1. When the antisense oligonucleotide was applied, the damage in CA3 and dentate gyrus was exaggerated dose-dependently, and this effect was more remarkable in CA3 than in the dentate gyrus. Based on these data, we concluded that: (i) a 72 kDa heat shock protein has a protective effect against the selective vulnerability of CA1 neurons, (ii) a 72 kDa heat shock protein is an essential factor for the tolerance exhibited by CA3 neurons, and (iii) dentate gyrus tolerance is based on mechanisms other than those mediated through a 72 kDa heat shock protein. PMID- 11927157 TI - Hippocampal alpha 7 and alpha 4 beta 2 nicotinic receptors and working memory. AB - Nicotine and other nicotinic receptor agonists have been found in a variety of studies to improve memory, while nicotinic receptor blockade can impair memory. The critical neural mechanisms for nicotinic involvement with memory are still under investigation. Initial evidence supports the involvement of the ventral hippocampus. Lesions in this area block nicotine-induced memory improvement and mecamylamine-induced impairment. Local ventral hippocampal application of the nicotinic channel blocker mecamylamine impairs memory in the 8-arm radial maze. Both alpha 4 beta 2 and alpha 7 nicotinic receptors seem to be involved. Ventral hippocampal infusions of high doses of the alpha 4 beta 2 nicotinic antagonist dihydro-beta-erythrodine (DH beta E) and the alpha 7 nicotinic antagonist methyllycaconitine (MLA) impair memory performance on the 8-arm radial maze. However, high doses of these drugs may limit specificity and they cause preconvulsant effects, which in themselves may affect memory. The current study used the more challenging 16-arm radial maze to determine the effects of lower doses of these drugs on memory and to differentiate effects on working and reference memory. Adult female Sprague-Dawley rats were trained on a working and reference memory task in the 16-arm radial maze and then were implanted with bilateral chronic guide cannulae directed to the ventral hippocampus. After recovery from surgery, the rats received acute intrahippocampal infusions of dose combinations of DH beta E and MLA. In the first study, DH beta E (0 and 6.75 microg/side) and MLA (0, 6.75, 13.5 and 27 microg/side) were administered in a counter-balanced order. In the second study, lower doses of DH beta E (0, 1.6375, 3.275 and 6.75 microg/side) were administered alone or with MLA (0 and 6.75 microg/side) in a counter-balanced order. In the first study, DH beta E caused a significant increase in both working and reference memory errors. MLA at a dose of 27 microg/side caused a significant increase in working memory errors, but this dose had no significant effect on reference memory errors. Interestingly, no additive effects were seen with combined administration of DH beta E and MLA in this study, and at the doses used, no effects were seen on response latency. In the second study, lower doses of DH beta E did not cause a significant deficit in working memory performance. Co-administration of MLA with these subthreshold doses did precipitate a memory impairment. The current results confirm the specificity of the memory deficits caused by these drugs. These results support the involvement of alpha 4 beta 2 and alpha 7 nicotinic receptors in the ventral hippocampus as being critical for memory function. PMID- 11927158 TI - Social experience and social context alter the behavioral response to centrally administered oxytocin in female Syrian hamsters. AB - The type of social behavior displayed by an individual is profoundly influenced by its immediate social environment or context and its prior social experience. Although oxytocin is important in the expression of social behavior in several species, it is not known if social factors alter the ability of oxytocin to influence behavior. The purpose of the present study was to test the hypothesis that social experience and social context alter the ability of oxytocin to regulate flank marking (a form of scent marking) in female Syrian hamsters. Oxytocin was microinjected into the medial preoptic anterior hypothalamic continuum (MPOA-AH) of socially experienced, dominant female hamsters which were then tested with either a subordinate partner, with a novel partner, or alone. Oxytocin induced flank marking in a dose-dependent manner but only when the experienced dominant hamsters were tested with their familiar, subordinate partners. Oxytocin did not induce flank marking when injected into socially naive female hamsters that were tested with an opponent or alone. In males, by contrast, oxytocin induced flank marking in dominant hamsters when they were tested with their subordinate partner or alone. These data support the hypothesis that social experience and social context interact to regulate the ability of oxytocin to stimulate flank marking by its actions in the MPOA-AH in female hamsters. PMID- 11927159 TI - GABA interacts with photic signaling in the suprachiasmatic nucleus to regulate circadian phase shifts. AB - Circadian rhythms of physiology and behavior in mammals are driven by a circadian pacemaker located in the suprachiasmatic nucleus of the hypothalamus. The majority of neurons in the suprachiasmatic nucleus are GABAergic, and activation of GABA receptors in the suprachiasmatic nucleus can induce phase shifts of the circadian pacemaker both in vivo and in vitro. GABA also modulates the phase shifts induced by light in vivo, and photic information is thought to be conveyed to the suprachiasmatic nucleus by glutamate. In the present study, we examined the interactions between GABA receptor agonists, glutamate agonists, and light in hamsters in vivo. The GABA(A) receptor agonist muscimol and the GABA(B) receptor agonist baclofen were microinjected into the suprachiasmatic nucleus at circadian time 13.5 (early subjective night), followed immediately by a microinjection of N methyl-D-aspartate (NMDA). Both muscimol and baclofen significantly reduced the phase shifting effects of NMDA. Further, coadministration of tetrodotoxin with baclofen did not alter the inhibition of NMDA by baclofen, suggesting a postsynaptic mechanism for the inhibition of NMDA-induced phase shifts by baclofen. Finally, the phase shifting effects of microinjection of muscimol into the suprachiasmatic nucleus during the subjective day were blocked by a subsequent light pulse. These data suggest that GABA regulates the phase of the circadian clock through both pre- and postsynaptic mechanisms. PMID- 11927161 TI - Vanilloid receptor 1 expression in the rat urinary tract. AB - Previous findings have shown that the capsaicin sensitivity of sensory fibres is due to the expression of a specific membrane protein, the vanilloid receptor type 1 (VR1). In the present work we studied the distribution, morphology and the neurochemical content of nerve fibres expressing this receptor in the rat urinary tract. Immunolabelling was performed against the VR1 and the positive fibres were examined by light and electron microscopy. Colocalisation of VR1 and substance P or calcitonin gene-related peptide immunoreactivities, and isolectin B4 binding, was evaluated under the confocal microscope. In addition, the effect of intravesical administration of resiniferatoxin, an ultra-potent vanilloid receptor agonist, in the receptor expression in the bladder was also studied. Numerous VR1-immunoreactive fibres were found in the mucosa and muscular layer of the entire urinary tract except the kidney. In the bladder, most fibres were also substance P- or calcitonin gene-related peptide-immunoreactive but did not bind isolectin B4. Under the electron microscope VR1 immunoreactivity was confined to unmyelinated axons and varicosities containing small clear and large dense-core synaptic vesicles. They occurred beneath or among epithelial cells or closely apposed to smooth muscle cells. Intravesical resiniferatoxin decreased VR1 immunoreactivity transiently. These data indicate that primary sensory fibres expressing VR1 are extremely abundant in the rat urinary tract and that, in contrast to the skin, they belong almost exclusively to the peptide-containing sub-population of primary afferents. As capsaicin-sensitive bladder afferents are involved in nociception and reflex micturition control, the numerous free terminal nerve endings expressing VR1 in the mucosa seem more adequate to accomplish the former function. However, the close apposition between VR1 expressing fibres and smooth muscle cells suggests that they may also encode the tonus of the muscular layer. PMID- 11927160 TI - Distribution of metabotropic glutamate receptors in the superior colliculus of the adult rat, ferret and cat. AB - The distribution of different metabotropic glutamate receptors (mGluRs 1a, 1b, 1c, 2/3, 4 and 5) has been compared in the superior colliculus of the rat, cat and ferret using immunohistochemical techniques and light microscopy. We found that although there are differences in labelling patterns between the species, there are also substantial similarities. In general, there was only light staining for the various mGluR1 splice variants, whereas labelling for the other Group I receptor, mGluR5, was heavier and with a pattern which suggested that at least some label arose from retinal afferents to the superficial superior colliculus. A further consistent feature in all species was labelling of astrocytes in the optic nerve/optic tract, superficial superior colliculus and brain at the collicular level with the antibody directed towards the Group II receptors, mGluR2 and mGluR3. Staining for the Group III receptor, mGluR4, was dense in the superficial superior colliculus in all species, with characteristics suggesting nerve fibre staining. mGluR4 staining was seen in the cat optic nerve/optic tract. One source of mGluR4 staining in the superior colliculus may thus be retinal axons, although other sources cannot be entirely excluded. These results demonstrate that distributions of mGluRs in these species have significant similarities but also some differences, suggesting that within the superior colliculus there may be some preservation of functional roles for some of the different receptor types. This is particularly so for the Group II and Group III receptors, which appear to have specific and distinct roles in the modulation of visual responses. PMID- 11927162 TI - Selective innervation of lamina I projection neurones that possess the neurokinin 1 receptor by serotonin-containing axons in the rat spinal cord. AB - Axons containing serotonin descend from brainstem to spinal cord and are thought to contribute to stimulation-produced and opioid analgesia, partly by a direct inhibitory action of serotonin on projection neurones. The density of serotoninergic innervation is highest in lamina I, which contains many nociceptive projection neurones. Two sets of anatomical criteria have been used to classify lamina I projection neurones: somatodendritic morphology and presence or absence of the neurokinin 1 receptor. To test whether the strength of serotoninergic innervation of lamina I projection neurones was related to morphology or neurokinin 1 receptor expression, we used confocal microscopy to determine the density of serotoninergic contacts on 60 cells retrogradely labelled from the caudal ventrolateral medulla. The contact density on neurones with the neurokinin 1 receptor was variable, with some cells receiving heavy input and others having few contacts. However, on average they received significantly more contacts (5.64 per 1000 microm(2) plasma membrane +/- 0.47, S.E.M.) than neurones which lacked the receptor (2.49 +/- .36). Among the neurokinin 1 neurones, serotoninergic innervation density was not related to morphology. Since the majority of serotoninergic boutons in lamina I of rat spinal cord do not appear to form synapses, we carried out electron microscopy on three heavily innervated neurokinin 1 receptor-immunoreactive projection neurones. Symmetrical synapses were found at 89% of serotoninergic contacts. These results indicate that serotoninergic innervation of lamina I projection neurones in the rat spinal cord is related to expression of neurokinin 1 receptors, but not to morphology, and that (at least on heavily innervated neurones) most serotonin-containing boutons which are in contact form synapses. PMID- 11927163 TI - Antihistamine antinociception is mediated by Gi-protein activation. AB - The effect of the i.c.v. administration of antisense oligodeoxynucleotides directed against the alpha subunit of different Gi-proteins (anti-Gi alpha(1), anti-Gi alpha(2), anti-Gi alpha(3)) on the antinociception induced by the H(1) antihistamines was evaluated in the mouse hot-plate test. The administration of diphenhydramine (20 mg kg(-1) s.c.), pyrilamine (15 mg kg(-1) s.c.) and promethazine (6 mg kg(-1) s.c.) produced an increase of the pain threshold which peaked 15 min after injection. Pretreatment with anti-Gi alpha(1) (12.5 microg per mouse i.c.v.), anti-Gi alpha(2) (25 microg per mouse i.c.v.) and anti-Gi alpha(3) (25 microg per mouse i.c.v.), administered 24 and 18 h before test, prevented the antihistamine-induced antinociception. At the highest effective doses, none of the compounds used impaired motor coordination, as revealed by the rota rod test, nor modified spontaneous motility and inspection activity, as revealed by the hole board test. These results suggest an important role played by the Gi-protein pathway in the transduction mechanism involved in the enhancement of the pain threshold produced by H(1)-antihistamines. PMID- 11927164 TI - Two alpha(2)-adrenergic receptor subtypes, alpha(2A) and alpha(2C), inhibit transmitter release in the brain of gene-targeted mice. AB - alpha(2)-Adrenergic receptors play an essential role in regulating neurotransmitter release from sympathetic nerves and from adrenergic neurons in the CNS. However, the role of each of the three highly homologous alpha(2) adrenergic receptor subtypes (alpha(2A), alpha(2B), alpha(2C)) in this process has not been determined unequivocally. To address this question, the regulation of norepinephrine and dopamine release was studied in mice carrying deletions in the genes encoding the three alpha(2)-adrenergic receptor subtypes. Autoradiography and radioligand binding studies showed that alpha(2)-receptor density in alpha(2A)-deficient brains was decreased to 9 +/- 1% of the respective wild-type value, whereas alpha(2)-receptor levels were reduced to 83 +/- 4% in alpha(2C)-deficient mice. These results indicate that approximately 90% of mouse brain alpha(2)-receptors belong to the alpha(2A) subtype and 10% are alpha(2C) receptors. In isolated brain cortex slices from wild-type mice a non-subtype selective alpha(2)-receptor agonist inhibited release of [(3)H]norepinephrine by maximally 96%. Similarly, release of [(3)H]dopamine from isolated basal ganglion slices was inhibited by 76% by an alpha(2)-receptor agonist. In alpha(2A) receptor-deficient mice, the inhibitory effect of the alpha(2)-receptor agonist on norepinephrine and dopamine release was significantly reduced but not abolished. Only in tissues from mice lacking both alpha(2A)- and alpha(2C) receptors was no alpha(2)-receptor agonist effect on transmitter release observed. The time course of onset of presynaptic inhibition of norepinephrine release was much faster for the alpha(2A)-receptor than for the alpha(2C) subtype. After prolonged stimulation with norepinephrine, presynaptic alpha(2C) adrenergic receptors were desensitized. From these data we suggest that two functionally distinct alpha(2)-adrenergic receptor subtypes, alpha(2A) and alpha(2C), operate as presynaptic inhibitory receptors regulating neurotransmitter release in the mouse CNS. PMID- 11927166 TI - Lamotrigine increases gene expression of GABA-A receptor beta3 subunit in primary cultured rat hippocampus cells. AB - To study the mechanism of action of the novel anticonvulsant and mood stabilizer lamotrigine, we used cDNA array to generate an expression profile of lamotrigine regulated genes in primary cultured rat hippocampus cells. Gene expression was analyzed using Atlas Rat 1.2 Arrays of approximately 1200 genes. Total RNA was isolated from control and cells treated with lamotrigine at 0.1 mM for one week and reverse-transcribed to cDNA labeled with 32P-dATP. The cDNA probes were hybridized with two identical cDNA array membranes. After comparing hybridized signals between these two membranes, we found that chronic treatment with lamotrigine increased the expression of eight genes and decreased the expression of six genes. One of the upregulated genes is GABA-A receptor beta subunit. This increase in GABA-A receptor beta3 subunit expression was further confirmed by Northern blotting analysis. In situ hybridization revealed that chronic treatment with lamotrigine increased GABA-A receptor beta3 subunit gene expression in CA1, CA3 and dentate gyrus of rat hippocampus. Our findings indicate that lamotrigine may effect GABA-A receptor regulated functions in the central nervous system. PMID- 11927165 TI - Lysophospholipids induce membrane hyperpolarization in microglia by activation of IKCa1 Ca(2+)-dependent K(+) channels. AB - Effects of the lysophospholipids sphingosine-1-phosphate and lysophosphatidic acid were studied in cultured murine microglia using the patch-clamp and video imaging techniques. Both lysophospholipids induced transient membrane hyperpolarization and K(+) current activation. The lysophospholipid-induced K(+) current was blocked by charybdotoxin or iberiotoxin, but was unaffected by apamin. In recordings with 1 microM intracellular free Ca(2+), Ca(2+)-dependent K(+) currents of microglia showed a similar pharmacological profile to lysophospholipid-induced currents. The Ca(2+)-dependent K(+) channels activated in microglia by lysophospholipids are most likely encoded by the IKCa1 channel gene. The presence of IKCa1 mRNA in microglia was demonstrated by reverse transcriptase-polymerase chain reaction studies. Ca(2+) imaging experiments revealed increases in the intracellular free Ca(2+) concentration of microglia to a mean value of about 400 nM after application of 1 microM sphingosine-1 phosphate or 1 microM lysophosphatidic acid. We suggest that the transient membrane hyperpolarization seen in microglia following exposure to sphingosine-1 phosphate or lysophosphatidic acid is caused by activation of IKCa1 Ca(2+) dependent K(+) channels. Increases in the concentration of intracellular free Ca(2+) evoked by the lysophospholipids are sufficient to activate microglial Ca(2+)-dependent K(+) channels. PMID- 11927167 TI - Dopaminergic mechanism for caffeine-produced cocaine seeking in rats. AB - Systemic administration of caffeine reinstates extinguished cocaine self administration behavior in rats, but the mechanism mediating this behavioral effect has not been established. The present study examined the role of adenosinergic A2 and dopaminergic mechanisms in caffeine-produced cocaine seeking. Following extinction of cocaine self-administration, experimenter administered injections of caffeine (1.25-20 mg/kg) and theophylline (1-10 mg/kg) dose-dependently reinstated extinguished cocaine-seeking behavior. Administration of the adenosinergic A2 antagonist, 3,7-dimethyl-1-propargylxanthine (DMPX; 0.546 2.18 microg/kg), failed to produce cocaine seeking. Pretreatment with doses of the adenosine A1/A2 agonist 5'-N-ethylcarboxamidoadenosine (NECA; 0.003-0.03 mg/kg) that were below those that produced marked sedation failed to block reinstatement. These data suggest that methylxanthine-produced cocaine seeking is not due to adenosine A2 receptor antagonism. In contrast, pretreatment with the dopaminergic D1-like antagonist SCH 23390 (0.005-0.02 mg/kg) or the D2-like antagonist eticlopride (0.03-0.3 mg/kg) produced a dose-dependent attenuation of caffeine-produced reinstatement at doses that did not decrease cocaine self administration. These findings suggest that dopaminergic mechanisms underlie the ability of caffeine to reinstate extinguished cocaine-taking behavior. PMID- 11927168 TI - Central nervous system monoamine correlates of social dominance in cynomolgus monkeys (Macaca fascicularis). AB - Social dominance is a fundamental component of both human and nonhuman primate sociality. However, its neurobiological correlates remain incompletely understood. We evaluated the association between dominance status and monoamine metabolite concentrations in cisternal cerebrospinal fluid (CSF) in adult male (n = 25) and female (n = 21) cynomolgus macaques (Macaca fascicularis) housed in unisexual social groups. Concentrations of the metabolites of dopamine (homovanillic acid [HVA]), norepinephrine (3-methoxy-4-hydroxyphenylglycol [MHPG]) and serotonin (5-hydroxyindoleacetic acid [5-HIAA]) were assayed. Dominant monkeys, both males and females, had significantly higher CSF HVA concentrations than did subordinates (p values <.05). Among males, but not females, dominants also had lower CSF 5-HIAA than subordinates (p <.05). The Dominance-HVA association observed here is consistent with recent speculation that social extraversion, a dominance-related personality trait in humans, may also reflect heightened central nervous system dopaminergic activity. PMID- 11927169 TI - Involvement of the sigma(1) receptor in cocaine-induced conditioned place preference: possible dependence on dopamine uptake blockade. AB - The involvement of the sigma(1) receptor on the rewarding effects of cocaine was examined using the conditioned place preference (CPP) procedure in C57BL/6 mice. Acquisition or expression of cocaine (20 mg/kg i.p.)-induced CPP was significantly decreased by pre-treatment with the selective sigma(1) receptor antagonists N,N-dipropyl-2-(4-methoxy-3-(2-phenylethoxy)phenyl)ethylamine (NE 100) or N-[2-(3,4-dichlorophenyl)ethyl]-N-methyl-2-(dimethylamino)ethylamine (BD1047), 1-10 mg/kg, i.p. The sigma(1) receptor agonists igmesine or 2-(4 morpholinoethyl-1-phenylcyclohexane-1-carboxylate hydrochloride (PRE-084) failed to induce CPP when injected alone. Moreover, the CPP induced by N-[1-(2 benzo(b)thiophenyl)cyclohexyl]piperidine (BTCP), a selective dopamine reuptake inhibitor, was blocked by treatments with the sigma(1) receptor antagonists as was similarly observed with cocaine. In addition, the repetitive treatment with cocaine during conditioning increased sigma(1) receptor mRNA expression in the nucleus accumbens, but not in the caudate putamen, prefrontal cortex or cerebellum. These data show that the sigma(1) receptor is not only necessary for acquisition of the cocaine-induced CPP, but that it is also implicated in its expression, confirming that activation of the sigma(1) receptor is induced during cocaine's early effects. The sigma(1) receptor is activated consequently to dopamine reuptake blockade and is not sufficient to induce CPP by itself. The mechanism of the sigma(1) receptor involvement in CPP and the selectivity toward the CPP-inducing drug remains however to be determined. These results show that strategies targeting the sigma(1) receptor with selective antagonists may allow effective attenuation of cocaine's rewarding properties and, in turn, offer new treatment strategies against drug addiction. PMID- 11927170 TI - Neuropharmacological profile of a novel potential atypical antipsychotic drug Y 931 (8-fluoro-12-(4-methylpiperazin-1-yl)- 6H-[1]benzothieno[2,3-b][1,5] benzodiazepine maleate). AB - The neuropharmacological profile of Y-931, 8-fluoro-12- (4-methylpiperazin-1-yl)- 6H-[1]benzothieno [2,3-b][1,5]benzodiazepine maleate, was investigated in comparison with those of typical and claimed atypical antipsychotic drugs. Similar to clozapine and olanzapine, Y-931 interacted with multiple neurotransmitter receptors such as dopaminergic, serotonergic, alpha-adrenergic, muscarinic and histaminergic receptors. Y-931, as well as the other antipsychotics, was active in a dose-dependent manner in established tests which are indicative of potential antipsychotic activity such as inhibition of apomorphine-induced hyperactivity and suppression of conditioned avoidance responses, however, only Y-931 and clozapine were devoid of cataleptogenic potential. In models of N-methyl-D-aspartate (NMDA) receptor hypofunction, Y-931 demonstrated the most potent protective action against the dizocilpine-induced neurotoxicity (neuronal vacuolization) in the rat retrosplenial cortex ([Y-931 (ED(50); 0.20 mg/kg, p.o.), olanzapine (1.1), clozapine (5.7), risperidone (6.9), haloperidol (19)). Furthermore, Y-931 and clozapine, unlike the other antipsychotics used, reversed the dizocilpine-induced social deficits at the same doses at which their neuroprotective action was exhibited. The present results suggest that Y-931 may be a novel potential atypical antipsychotic drug with a low risk of extrapyramidal syndrome (EPS) and the property to ameliorate NMDA receptor hypofunction. PMID- 11927171 TI - Effects of age, postmortem delay and storage time on receptor-mediated activation of G-proteins in human brain. AB - The influence of age, postmortem delay and freezing storage period on receptor mediated G-protein activity was quantified in cortical membranes from 34 healthy subjects. Concentration-response curves of the [(35)S]GTPgammaS binding stimulation by agonists for alpha(2)-adrenoceptors (UK14304), mu-opioid (DAMGO), 5-HT(1A) (8-OH-DPAT), GABA(B) (baclofen) and muscarinic (carbachol) receptors were analyzed. Immunoreactivities of G(alpha)-protein subunits were also determined. Basal binding and UK14304, 8-OH-DPAT, and baclofen potency to stimulate [(35)S]GTPgammaS binding decreased with aging (1-88 years) without changes of efficacy. DAMGO-mediated stimulation increased both in potency and efficacy with aging. A negative correlation between age and immunoreactivity was observed for G(alphai1/2)-, but not for G(alphai3)-, G(alphao)-,and G(alphas) proteins. Neither [(35)S]GTPgammaS binding nor G(alpha)-proteins changed with the postmortem delay (8-92 h). Basal [(35)S]GTPgammaS binding decreased with the sample storage period (1-85 months). A careful match between cases and controls should be taken into account when designing signal transduction studies in human disorders, specially for variables such as age and storage period. PMID- 11927172 TI - From first drug use to drug dependence; developmental periods of risk for dependence upon marijuana, cocaine, and alcohol. AB - The focal point of this paper is the transition from drug use to drug dependence. We present new evidence on risk for starting to use marijuana, cocaine, and alcohol, as well as risks for progression from first drug use to the onset of drug dependence, separately for each of these drugs. Data from the National Comorbidity Survey (NCS) were analyzed. The NCS had a representative sample of the United States population ages 15-54 years (n = 8,098). Survival analysis techniques were used to provide age- and time-specific risk estimates of initiating use of marijuana, cocaine, and alcohol, as well as of becoming dependent on each drug. With respect to risk of initiating use, estimated peak values for alcohol and marijuana were found at age 18, about two years earlier than the later peak in risk of initiating cocaine use. With respect to risk of meeting criteria for the clinical dependence syndrome, estimated peak values for alcohol and marijuana were found at age 17-18. Peak values for cocaine dependence were found at age 23-25. Once use began, cocaine dependence emerged early and more explosively, with an estimated 5-6% of cocaine users becoming cocaine dependent in the first year of use. Most of the observed cases of cocaine dependence met criteria for dependence within three years after initial cocaine use. Whereas some 15-16% of cocaine users had developed cocaine dependence within 10 years of first cocaine use, the corresponding values were about 8% for marijuana users, and 12-13% for alcohol users. The most novel findings of this study document a noteworthy risk for quickly developing cocaine dependence after initial cocaine use, with about one in 16 to 20 cocaine users becoming dependent within the first year of cocaine use. For marijuana and alcohol, there is a more insidious onset of the drug dependence syndrome. PMID- 11927173 TI - Identification of a naturally occurring polymorphism in the promoter region of the norepinephrine transporter and analysis in major depression. AB - Disturbances in the noradrenergic neurotransmission system have been implicated in the etiology of mood disorders. The norepinephrine transporter (NET) is a main target of antidepressant action and was shown to be dysregulated in major depression. Despite the clinical and physiological significance of NET gene regulation, little is known about the transcriptional control mechanisms governing its expression. Since it is well established that affective disorders have a genetic component with many genes of small effect contributing to the genetic susceptibility of depression, the NET gene is an interesting candidate gene for affective disorders. In a search for polymorphisms or mutations in the 5' flanging region of the NET gene we sequenced approximately 1000 bp upstream of the first codon in the NET gene promoter in 100 patients with major depression and 100 healthy controls. We identified a so far unknown T --> C polymorphism 182 bp upstream of the start codon in a transcriptional relevant region. In a case control association study we investigated the newly identified T-182C polymorphism and an already known G1287A polymorphism in exon 9 of the NET gene in a sample of 193 patients with major depression and 136 healthy, non-related controls. No statistical significant differences between patients and controls were found for any of the analyzed polymorphisms, either in the genotype distribution or in the allele frequencies. Our results suggest that the investigated polymorphisms are not major susceptibility factors in the etiology of major depression. PMID- 11927174 TI - Comparison of rapidly acting intramuscular olanzapine, lorazepam, and placebo: a double-blind, randomized study in acutely agitated patients with dementia. AB - This double-blind study investigated the efficacy and safety of rapid-acting intramuscular olanzapine in treating agitation associated with Alzheimer's disease and/or vascular dementia. At 2 h, olanzapine (5.0 mg, 2.5 mg) and lorazepam (1.0 mg) showed significant improvement over placebo on the PANSS Excited Component (PANSS-EC) and Agitation-Calmness Evaluation Scale (ACES), and both 5.0 mg olanzapine and lorazepam showed superiority to placebo on the Cohen Mansfield Agitation Inventory. At 24 h, both olanzapine groups maintained superiority over placebo on the PANSS-EC; lorazepam did not. Olanzapine (5.0 mg) and lorazepam improved ACES scores more than placebo. Simpson-Angus and Mini Mental State Examination scores did not change significantly from baseline. Sedation (ACES > or =8), adverse events, and laboratory analytes were not significantly different from placebo for any treatment. No significant differences among treatment groups were seen in extrapyramidal symptoms or in corrected QT interval at either 2 h or 24 h, and no significant differences among treatment groups were seen in vital signs, including orthostasis. Intramuscular injection of olanzapine may therefore provide substantial benefit in rapidly treating inpatients with acute dementia-related agitation. PMID- 11927175 TI - The effects of hydrocortisone on cognitive and neural function: a behavioral and event-related potential investigation. AB - Research indicates that glucocorticoids affect hippocampal function and the form of cognition that the hippocampus is thought to subserve, explicit memory. However, some studies suggest that glucocorticoids affect the frontal lobe, attention and working memory. Thus, it is not clear whether glucocorticoids specifically target the hippocampus and explicit memory or if the effects are more ubiquitous. By simultaneously measuring event-related potentials and behavioral performance in tasks designed to tap particular cognitive and neural processes, the present study examined the effects of hydrocortisone on 24 healthy humans. In an intentional face recognition memory task where the stimuli were presented again after a brief delay (6-18 s) and a long delay (30 min), hydrocortisone altered the P600 component (an electrophysiological index of recognition memory and hippocampal activity) following the brief delay and impaired behavioral performance after the long delay. ERPs and behavioral performance were not affected in the attention and working memory tasks. These findings are consistent with reports indicating that glucocorticoids affect explicit memory and hippocampal function. PMID- 11927176 TI - Marijuana effects on sensitivity to reinforcement in humans. AB - Under controlled laboratory conditions, eight adult subjects smoked placebo and three different potencies of marijuana cigarettes ranging in Delta(9) THC content. Immediately following smoking, subjects were exposed to a laboratory task that provided concurrently available response options. One option systematically decreased in reinforcement frequency throughout the session, and thus required a reallocation of behavior to the non-decreasing option to maximize monetary earnings. After smoking the two highest doses (1.77% and 3.58% Delta(9) THC) subjects earned fewer reinforcers and allocated a higher proportion of responding to the decreasing option, compared with placebo and the lowest dose. The difference in reinforcers earned could not be accounted for by a change in response rates. Quantitative and graphical analyses revealed that the higher doses produced considerable periods of time spent on the decreasing option despite earning few reinforcers. The data are discussed with regard to marijuana effects on dopamine/cannabinoid systems and adaptive behavior change. PMID- 11927177 TI - Pattern of response to divalproex, lithium, or placebo in four naturalistic subtypes of mania. AB - We investigated effects of antimanic treatments on specific aspects of mania, prediction of response, and the existence of naturalistic subgroups of patients with different treatment response in 179 inpatients randomized to antimanic treatment with lithium, divalproex, or placebo. Psychiatric symptom ratings were conducted by clinicians and nurses before and during treatment. Factor analysis using physician and nurse rating scales, followed by a cluster analysis, yielded anxious-depressive, psychotic, classic, and irritable subtypes. We compared: (1) treatment effects on factor scores; (2) responses to treatment across subtypes; and (3) pattern of symptom change with each treatment. The anxious-depressed subtype did not respond to any treatment; the psychotic and classic subtypes responded similarly to lithium and to divalproex; and the irritable-dysphoric subtype responded better to divalproex than to lithium. Overall, divalproex improved impulsivity and hostility significantly more than placebo, and lithium or divalproex improved hyperactivity more than placebo. These data suggest that there are naturalistic subtypes of manic episodes with different responses to treatment. PMID- 11927178 TI - Novel factor-based symptom scores in treatment resistant schizophrenia: implications for clinical trials. AB - To study the factor structure of symptoms in patients with treatment resistant schizophrenia and whether it is altered by treatment, we analyzed ratings on the Brief Psychiatric Rating Scale (BPRS) from two independent groups of patients with treatment resistant schizophrenia. With confirmatory factor analysis of pre clozapine BPRS scores in 1074 patients in an administrative data base, the Clozapine Authorization and Monitoring Program (CAMP), we assessed the fit of published factor models and developed a better-fitting model. Model fit was validated in an independent group of 197 research unit participants. Stability of model fit six months post-clozapine was assessed in 834 CAMP patients. A new 4 factor model (negative symptoms, reality distortion, disorganization, and anxiety/depression) had better fit in both data sets than two commonly used factor models, and also fit better post-clozapine. We recommend these four factor scores as clinical trial outcomes in patients with treatment resistant schizophrenia. PMID- 11927179 TI - 1H MRS brain measures and acute lorazepam administration in healthy human subjects. AB - The effects of acute lorazepam administration on 1H magnetic resonance spectroscopy (MRS) in vivo brain spectra were examined in the left dorsolateral prefrontal cortex (L-DLPFC) of healthy human subjects. We wanted to examine whether lorazepam administration would result in significant changes in the levels of 1H-MRS metabolites in this brain region. Ten healthy controls underwent a short echo-time 1H-MRS session immediately before, and a second one 1 h after lorazepam administration (2mg/orally). The measured 1H-metabolites included N acetyl-aspartate, phosphocreatine+creatine, trimethylamines, myo-inositol, glutamate, and glutamine, which were expressed as absolute values and ratios. No significant differences were found after lorazepam administration for any of the measured metabolite levels or ratios (paired t-tests, p >.05). This study demonstrated that lorazepam can potentially be utilized to acutely sedate psychiatric subjects during in vivo 1H-MRS sessions, as it does not appear to produce significant changes in the 1H-MRS spectra in this specific brain region. PMID- 11927180 TI - The selective serotonin(2A) receptor antagonist, MDL100,907, elicits a specific interoceptive cue in rats. AB - Employing a two-lever, food-reinforced, Fixed Ratio 10 drug discrimination procedure, rats were trained to recognize the highly-selective serotonin (5 HT)(2A) receptor antagonist, MDL100,907 (0.16 mg/kg, i.p.). They attained criterion after a mean +/- S.E.M. of 70 +/- 11 sessions. MDL100,907 fully generalized with an Effective Dose (ED)(50) of 0.005 mg/kg, s.c. A further selective 5-HT(2A) antagonist, SR46349, similarly generalized with an ED(50) of 0.04 mg/kg, s.c. In distinction, the selective 5-HT(2B) antagonist, SB204,741 (0.63 and 10.0 mg/kg), the 5-HT(2B/2C) antagonist, SB206,553 (0.16 and 2.5 mg/kg) and the selective 5-HT(2C) antagonists, SB242,084 (2.5 and 10.0 mg/kg,) and RS102221 (2.5 and 10.0 mg/kg), did not significantly generalize. In conclusion, selective blockade of 5-HT(2A) receptors by MDL100,907 elicits a discriminative stimulus in rats which appears to be specifically mediated via 5-HT(2A) as compared with 5-HT(2B) and 5-HT(2C) receptors. PMID- 11927181 TI - Differential regulation of 5-HT1A receptor-G protein interactions in brain following chronic antidepressant administration. AB - Changes in 5-HT(1A) receptor function or sensitivity following chronic antidepressant treatment may involve changes in receptor-G protein interaction. We have examined the effect of chronic administration of the SSRI fluoxetine or the tricyclic antidepressant amitriptyline on 5-HT(1A) receptor-stimulated [(35)S]GTPgammaS binding in serotonergic cell body areas, and cortical and limbic structures using quantitative autoradiography. Treatment of rats with fluoxetine, but not amitriptyline, resulted in an attenuation of 5-HT(1A) receptor-stimulated [(35)S]GTPgammaS binding in the dorsal and median raphe nuclei. The binding of the antagonist radioligand [3H]MPPF to 5-HT(1A) receptor sites was not altered, suggesting that the observed changes in 5-HT(1A) receptor-stimulated [(35)S]GTPgammaS binding were not due to changes in receptor number. Thus, the desensitization of somatodendritic 5-HT(1A) autoreceptors in the dorsal and median raphe following chronic SSRI treatment appears to be due to a reduced capacity of the 5-HT(1A) receptor to activate G protein. By contrast, no significant change in postsynaptic 5-HT(1A) receptor-stimulated [(35)S]GTPgammaS binding was observed in any of the forebrain areas examined following chronic antidepressant treatment. Thus, changes in postsynaptic 5-HT(1A) receptor mediated responses reported to follow chronic SSRI or tricyclic antidepressant administration most likely occur distal to receptor-G protein interaction, perhaps at the level of effector, or involving changes in neuronal function at the system or circuit level. PMID- 11927182 TI - Effects of a non-peptide CRF antagonist (DMP696) on the behavioral and endocrine sequelae of maternal separation. AB - We examined whether blockade of corticotropin-releasing factor (CRF) receptors by a non-peptide CRF antagonist (DMP696) would attenuate the stress hyper responsiveness that occurs in response to maternal separation. In a social interaction test as well as the elevated plus maze, adult male rats, which had been maternally separated as infants, displayed more anxiety-like behavior compared with handled rats. DMP696 increased social interaction in both groups. In the elevated plus maze however, DMP696 significantly increased open arm time in the maternally separated rats but not in the handled group whereas chlordiazepoxide increased open arm time in both groups. DMP696 also appeared to block stress-induced ACTH secretion more readily in the maternally separated group compared with the handled rats. These observations suggest that CRF antagonists are particularly effective in animals that are hyper-responsive to stress and may therefore have utility in the treatment of anxiety and affective disorders where CRF has been implicated. PMID- 11927184 TI - Effects of haloperidol and risperidone on neurotensin levels in brain regions and neurotensin efflux in the ventral striatum of the rat. AB - Neurotensin (NT) may play a role in the pathophysiology of schizophrenia and in the mechanism of action of antipsychotic drugs. Here we studied the effects of a 30-day regimen of haloperidol (1.15 mg/100 g food) and risperidone (1.15 and 2.3 mg/100 g food) on NT-like immunoreactivity (-LI) levels in brain tissue and NT-LI efflux in the ventral striatum (VSTR) of the rat. Haloperidol, but not risperidone, increased NT-LI levels in the striatum. In the occipital cortex, risperidone, but not haloperidol, decreased levels of NT-LI. In the hippocampus and the frontal cortex both haloperidol and risperidone (the higher dose) increased NT-LI levels. In the VSTR, haloperidol and risperidone (the higher dose) decreased NT-LI efflux and abolished the stimulatory effect of d amphetamine (1.5 mg/kg, s.c.). Thus, changes in NT occur in response to antipsychotic drugs and psychostimulants that may be relevant for the pathophysiology and treatment of schizophrenia. PMID- 11927183 TI - Effects of typical and atypical antipsychotics and receptor selective compounds on acetylcholine efflux in the hippocampus of the rat. AB - Some atypical antipsychotic drugs appear to improve cognitive function in schizophrenia and since acetylcholine (ACh) is of importance in cognition, we used in vivo microdialysis to examine the effects of antipsychotics administered acutely (SC or IP) at pharmacologically comparable doses on ACh outflow in the hippocampus of the rat. The atypical antipsychotics olanzapine and clozapine produced robust increases in ACh up to 1500% and 500%, respectively. The neuroleptics haloperidol, thioridazine, and chlorpromazine, as well as the atypical antipsychotics risperidone and ziprasidone produced modest increases in ACh by about 50-100%. Since most atypical antipsychotics affect a variety of monoaminergic receptors, we examined whether selective ligands for some of these receptors affect hippocampal ACh. Antagonists for the 5-HT(2A) (MDL 100,907), the 5-HT(2C) (SB 242,084), the 5-HT(6) (Ro 04-6790), the D(2) (raclopride) receptors, and the alpha(1)-adrenoceptors (prazosin) modestly increased ACh by about 50%. The 5-HT(1A) agonist R-(+)-8-OH-DPAT and the alpha(2)-adrenoceptor antagonist yohimbine significantly increased ACh by about 100% and 50%, respectively. Thus, olanzapine and clozapine increased ACh to a greater extent than other tested antipsychotics, explaining perhaps their purported beneficial effect in cognitive function in schizophrenia. It appears that selective activity at each of the monoaminergic receptors studied is not the sole mechanism underlying the olanzapine and clozapine induced increases in hippocampal ACh. PMID- 11927185 TI - Altered regulation of CREB by chronic antidepressant administration in the brain of transgenic mice with impaired glucocorticoid receptor function. AB - Various effects of antidepressant drugs on gene transcription have been described and altered gene expression has been proposed as being a common biological basis underlying depressive illness. One target for the common action of antidepressants is a modifying effect on the regulation of postreceptor pathways and genes related to the cAMP cascade. Recent studies have demonstrated that long term antidepressant treatment resulted in sustained activation of the cyclic adenosine 3',5'-monophosphate system and in increased expression of the transcription factor cAMP response element binding protein (CREB). A transgenic animal model of depression with impaired glucocorticoid receptor function was used to investigate the effect of chronic antidepressant treatments on CREB expression in different brain areas. Wild-type and transgenic mice received one administration of saline, desipramine, or fluoxetine, daily for 21 days. The effects of antidepressants on CREB mRNA were analyzed using a sensitive RNase protection assay. Antidepressant treatment resulted in a neuroanatomically and animal specific expression pattern of CREB. Our findings suggest that life-long central glucocorticoid receptor dysfunction results in an altered sensitivity with respect to the effects of antidepressants on the expression of CREB. PMID- 11927186 TI - The neuropeptide Y (NPY) Y1 receptor subtype mediates NPY-induced antidepressant like activity in the mouse forced swimming test. AB - The present study was undertaken to investigate the possible antidepressant-like effects of neuropeptide Y (NPY) in the mouse forced swimming test, an animal model widely used for the screening of potential antidepressant drugs. In addition, experiments were performed, using agonists and selective antagonists, to assess the potential role of NPY Y(1) and Y(2) receptor subtypes in this model. Complementary studies were performed in an open field apparatus to rule out any changes in locomotor activity that might have interfered with the interpretation of data from the mouse forced swimming test. Intracerebroventricular injections (0.03 nmole-3 nmole) of NPY, [Leu(31)Pro(34)]PYY (Y(1) agonist), NPY(13-36) (Y(2) agonist), BIBP3226, BIBO3304 (Y(1) antagonists) and BIIE0246 (Y(2) antagonist) were performed 30 min prior to testing in the mouse forced swimming test and open field. NPY administration significantly reduced immobility time in a dose dependent manner (p <.01 vs. control group), as did [Leu(31)Pro(34)]PYY (p <.01 vs. control group) and BIIE0246 (p <.05 vs. control group). In contrast, BIBO3304, BIBP3226 and NPY(13 36) did not display any activity at the doses tested. However, pretreatment with BIBO3304 or BIBP3226 significantly blocked the anti-immobility effects of NPY. Data from the open field demonstrated that BIIE0246 increased horizontal ambulation at the dose found to be active in the forced swimming test. Taken together, our results demonstrate that NPY displays antidepressant-like activity in the mouse forced swimming test, and suggest that this activity is mediated by the NPY Y(1) receptor subtype. PMID- 11927187 TI - Plasticity of dopamine D4 receptors in rat forebrain: temporal association with motor hyperactivity following neonatal 6-hydroxydopamine lesioning. AB - Genetic studies suggest that dopamine D(4) receptor polymorphism is associated with attention deficit hyperactivity disorder (ADHD). We recently reported that motor hyperactivity in juvenile male rats with neonatal 6-hydroxydopamine lesions of the central dopamine system can be reversed by dopamine D(4) receptor selective antagonists. In this study, effects of such lesions on D(4) as well as other dopamine receptors (D(1) and D(2)) were autoradiographically quantified at selected developmental stages. Neonatal lesions resulted in motor hyperactivity at postnatal day (PD) 25, but not at PD 37 or 60. Correspondingly, D(4) receptor levels in lesioned rats were substantially increased in caudate-putamen and decreased in nucleus accumbens at PD 25, but not at PD 37 or 60. Neonatal lesions also led to relatively minor changes in D(1) and D(2) receptor binding in various forebrain regions. However, the time-course of lesion-induced motor hyperactivity correlated only with changes in D(4), but not D(1) and D(2) receptors. These results further support the hypothesis that D(4) receptors may play a pivotal role in lesion-induced hyperactivity, and possibly in clinical ADHD. PMID- 11927188 TI - A single dose of lysergic acid diethylamide influences gene expression patterns within the mammalian brain. AB - Hallucinogenic drugs such as lysergic acid diethylamide (LSD) have profound effects on humans including hallucinations and detachment from reality. These remarkable behavioral effects have many similarities to the debilitating symptoms of neuropsychiatric disorders such as schizophrenia. The effects of hallucinogens are thought to be mediated by serotonin receptor activation; however, how these drugs elicit the unusual behavioral effects remains largely a mystery, despite much research. We have undertaken the first comprehensive analysis of gene expression influenced by acute LSD administration in the mammalian brain. These studies represent a novel approach to elucidate the mechanism of action of this class of drugs. We have identified a number of genes that are predicted to be involved in the processes of synaptic plasticity, glutamatergic signaling and cytoskeletal architecture. Understanding these molecular events will lead to new insights into the etiology of disorders whose behavioral symptoms resemble the temporary effects of hallucinogenic drugs, and also may ultimately result in new therapies. PMID- 11927189 TI - Neurobehavioral effects of interferon-alpha in cancer patients: phenomenology and paroxetine responsiveness of symptom dimensions. AB - We have previously shown that the risk of major depression in patients with malignant melanoma undergoing interferon-alpha (IFN-alpha) therapy can be reduced by pretreatment with the antidepressant, paroxetine. Using dimensional analyses, the present study assessed the expression and treatment responsiveness of specific clusters of neuropsychiatric symptoms over the first three months of IFN alpha therapy. Forty patients with malignant melanoma eligible for IFN-alpha treatment were randomly assigned to receive either paroxetine or placebo in a double-blind design. Neuropsychiatric assessments were conducted at regular intervals during the first twelve weeks of IFN-alpha therapy and included the 21 item Hamilton Depression Rating Scale, the 14-item Hamilton Anxiety Rating Scale and the Neurotoxicity Rating Scale. Neurovegetative and somatic symptoms including anorexia, fatigue and pain appeared within two weeks of IFN-alpha therapy in a large proportion of patients. In contrast, symptoms of depressed mood, anxiety and cognitive dysfunction appeared later during IFN-alpha treatment and more specifically in patients who met DSM-IV criteria for major depression. Symptoms of depression, anxiety, cognitive dysfunction and pain were more responsive, whereas symptoms of fatigue and anorexia were less responsive, to paroxetine treatment. These data demonstrate distinct phenomenology and treatment responsiveness of symptom dimensions induced by IFN-alpha, and suggest that different mechanisms mediate the various behavioral manifestations of cytokine induced "sickness behavior." PMID- 11927191 TI - Laboratory-measured aggressive behavior of women: acute tryptophan depletion and augmentation. AB - Plasma L-tryptophan (Trp) reductions have been related to aggression increases in men. Impairment of serotonin synthesis and neurotransmission is one explanation. Using repeated-measures, this Trp manipulation study measured laboratory-induced aggression in 12 women after Trp augmentation (T+), depletion (T-), and food restricted (fasting control) conditions. Participants were provoked with periodic subtraction of money from their task earnings by a (fictitious) partner. Aggression was defined as the number of point subtractions participants made from their fictitious partner. Participants completed five testing sessions under each condition. T+ decreased aggressive responses and T- increased aggressive responses. Post-hoc analyses showed changes in aggressive behavior were specific to women with higher fasting control plasma Trp, which is consistent with research demonstrating that men with higher levels of baseline Trp are more aggressive. These findings indicate that both T+ and T- can influence aggressive behavior and that certain subgroups of women may be more susceptible to serotonin manipulation. PMID- 11927190 TI - A longitudinal study of serotonergic function in depression. AB - Several reports have described abnormal neuroendocrine responses to serotonergic challenge tests in depression, but few have studied depressed patients followed longitudinally. In order to determine whether blunted prolactin responses to clomipramine challenge is a "state" vs. "trait" marker in depression, we applied this challenge paradigm to 20 patients with Major Depression prior to treatment and at three additional time points following response to desipramine: at the completion of acute treatment; at the end of the continuation phase of treatment; and after a minimum of three weeks "washout" following the discontinuation of treatment. The prolactin response to clomipramine challenge was blunted in depressed patients compared with matched healthy control subjects, at each time point over the longitudinal course of their illness and recovery. Challenge test results in depressed patients did not change after response to acute desipramine therapy, at the conclusion of the continuation phase of treatment, or while in a medication-free state of remission. Blunted prolactin response to clomipramine challenge persists in depressed patients after recovery from acute illness, and may reflect an underlying biological vulnerability. PMID- 11927192 TI - Sexual diergism of hypothalamo-pituitary-adrenal cortical responses to low-dose physotigmine in elderly vs. young women and men. AB - We previously demonstrated that the reversible cholinesterase inhibitor, physostigmine (PHYSO), administered to normal young adult women and men (average age 35 years) at a dose that produced few or no side effects, resulted in a sex difference (sexual diergism) in hypothalamo-pituitary-adrenal cortical (HPA) axis responses: Plasma ACTH(1-39), cortisol, and arginine vasopressin (AVP) concentrations increased to a significantly greater extent in the men than in the women. To explore the effect of age on these sexually diergic hormone responses, in the present study we used the same dose of PHYSO (8 microg/kg IV) to stimulate ACTH(1-39), cortisol, and AVP secretion in normal elderly, non-estrogen-replaced women and elderly men (average ages 73 years and 70 years, respectively). The subjects underwent three test sessions 5-7 days apart: PHYSO, saline control, and a second session of PHYSO. Serial blood samples were taken for hormone analyses before and after pharmacologic challenge. As with the previously studied younger subjects, PHYSO administration produced no side effects in about half the elderly subjects and mild side effects in the other half, with no significant female-male differences. The hormone responses were 2-5 fold greater in the elderly subjects than in the younger subjects, but in contrast to the younger subjects, the elderly men did not have significantly greater hormone responses to PHYSO administration than did the elderly women. The ACTH(1-39) and AVP responses to PHYSO for the two sessions were significantly positively correlated in the men (+0.96, +0.91) but not in the women. None of the hormone responses was significantly correlated with the presence or absence of side effects in either group of subjects.These results indicate a greater sensitivity of the HPA axis to low-dose PHYSO, and a loss of overall sex differences in hormone responses, in elderly compared with younger subjects. The lack of a difference in side effects between the elderly women and men and the lack of significant correlations between presence or absence of side effects and hormone responses suggest that the increase in hormone responses with aging is due to correspondingly increased responsiveness of central cholinergic systems and/or the HPA axis, and not to a nonspecific stress response. PMID- 11927193 TI - Decision-making in a risk-taking task: a PET study. AB - As decision-making is central to motivated behavior, understanding its neural substrates can help elucidate the deficits that characterize various maladaptive behaviors. Twenty healthy adults performed a risk-taking task during positron emission tomography with (15)O-labeled water. The task, a computerized card game, tests the ability to weigh short-term rewards against long-term losses. A control task matched all components of the risk-taking task except for decision-making and the difference between responses to contingent and non-contingent reward and punishment. Decision-making (2 runs of the active task minus 2 runs of the control task) activated orbital and dorsolateral prefrontal cortex, anterior cingulate, insula, inferior parietal cortex and thalamus predominantly on the right side, and cerebellum predominantly on the left side. In an exploratory analysis, guessing (run 1 minus run 2 of the active task) accompanied activation of sensory-motor associative areas, and amygdala on the left side, whereas informed decision-making (run 2 minus run 1) activated areas that subserve memory (hippocampus, posterior cingulate) and motor control (striatum, cerebellum). The findings provide a framework for future investigations of decision-making in maladaptive behaviors. PMID- 11927194 TI - A gender-specific association between the serotonin transporter gene and suicide attempts. AB - This pilot study tested the gender-specificity of the association between suicide attempts and a polymorphism in the promoter area of the serotonin transporter with two allelic variants, a long (1) variant and a short (s) variant. In a Spanish general hospital, 180 suicide attempters (121 women and 59 men) and 212 control blood donors (93 women and 119 men) were recruited. Subjects were classified as S individuals (s/s or s/l) with low expression of the serotonin transporter, and L individuals (l/l) with high expression. S individuals were significantly overrepresented (or L, underrepresented) in female attempters when compared with female controls and male attempters. Lethality appeared to have a significant influence on the effects of the genotype in suicide since S females were overrepresented among non-lethal female attempters. Further studies are needed to replicate that the serotonin genotype polymorphism may influence suicide attempts only in females. PMID- 11927195 TI - Improving scientific publications and public trust by data access. PMID- 11927196 TI - The tyramine pressor test may have limited sensitivity, especially in the presence of dual serotonin/norepinephrine uptake inhibition. PMID- 11927198 TI - Attrition in the Longitudinal Aging Study Amsterdam. The effect of differential inclusion in side studies. AB - This study addresses the relation between attrition and characteristics of the study protocol, specifically contact frequency, and respondent burden. The study is based on data from a longitudinal study with side studies on various topics, so that respondents have differential exposure to these study characteristics. Attrition outcomes are refusal and ineligibility through frailty. The effect of side study contact frequency and respondent burden on these outcomes is examined in two analytical samples: (1) baseline participants surviving to the first follow-up after 10 months (sample I), and (2) first follow-up participants surviving to the second follow-up after 3 years (sample II). Attrition during the first study interval was higher than during the second study interval, 15.5 and 5.4%, respectively. In sample I, the request to participate in a side study on social network implied an increased risk of refusal to participate at first follow-up if subjects refused the request (RR 8.34). However, if subjects participated in the network study, their risk of refusal was decreased (RR 0.42). In sample II, requests to participate in one to four side study cycles increased the risk of refusal to participate at second follow-up if subjects participated in fewer cycles than requested (RR 9.21). If subjects participated in all side study cycles that they were approached for, even if the number of cycles was five or more, this had an opposite effect: it decreased the risk of refusal (RR 0.18). Ineligibility was not significantly associated with contact frequency or respondent burden. Furthermore, neither contact frequency nor respondent burden related refusal was selective with respect to socio-demographic characteristics and physical and mental health indicators. It is concluded that contact frequency is nonlinearly associated with attrition. The findings further suggest that designing a series of side studies within the "longitudinal paradigm" does not severely damage the study's validity in terms of selective attrition. PMID- 11927199 TI - Attrition in longitudinal studies. How to deal with missing data. AB - The purpose of this paper was to illustrate the influence of missing data on the results of longitudinal statistical analyses [i.e., MANOVA for repeated measurements and Generalised Estimating Equations (GEE)] and to illustrate the influence of using different imputation methods to replace missing data. Besides a complete dataset, four incomplete datasets were considered: two datasets with 10% missing data and two datasets with 25% missing data. In both situations missingness was considered independent and dependent on observed data. Imputation methods were divided into cross-sectional methods (i.e., mean of series, hot deck, and cross-sectional regression) and longitudinal methods (i.e., last value carried forward, longitudinal interpolation, and longitudinal regression). Besides these, also the multiple imputation method was applied and discussed. The analyses were performed on a particular (observational) longitudinal dataset, with particular missing data patterns and imputation methods. The results of this illustration shows that when MANOVA for repeated measurements is used, imputation methods are highly recommendable (because MANOVA as implemented in the software used, uses listwise deletion of cases with a missing value). Applying GEE analysis, imputation methods were not necessary. When imputation methods were used, longitudinal imputation methods were often preferable above cross-sectional imputation methods, in a way that the point estimates and standard errors were closer to the estimates derived from the complete dataset. Furthermore, this study showed that the theoretically more valid multiple imputation method did not lead to different point estimates than the more simple (longitudinal) imputation methods. However, the estimated standard errors appeared to be theoretically more adequate, because they reflect the uncertainty in estimation caused by missing values. PMID- 11927200 TI - Accuracy of blood pressure measurement and anthropometry among volunteer observers in a large community survey. AB - The Busselton Survey is a population survey that is held about every three years. In 1994-1995 a re-survey was held of all past participants and 8,502 attended. Financial constraints precluded employing staff for data collection for blood pressure and anthropometry, these therefore were collected by unpaid lay volunteers. Quality control by a health professional was critical to the assessment and maintenance of accuracy in these measurements. For blood pressure three readings were taken simultaneously by a quality control person and the volunteer using a dual stethoscope. Duplicate anthropometric measurements were taken by a criterion anthropometrist and the volunteer. Inter and intra-observer technical errors of measurement (TEM) were calculated. Sixty-two volunteers were trained to take BP measurements; of these, 38 collected data, and 63 were trained in anthropometry; of these, 30 were suitable as measurers. Training was conducted on a group and individual basis by the quality assurance person for the study both in the Perth metropolitan area and rural Busselton. The TEM for SBP was 1.6 mm Hg (SD 1.0 mm Hg) and 1.5 mm Hg (SD 0.8) for DBP. For skin-folds intra observer TEM ranged from 0.6 mm to 1.0 mm. Between-observer TEM ranged from 2.1 mm to 5.4 mm. For limb and waist circumferences, intra-observer TEM ranged from 0.3 cm to 1.6 cm. Between-observer TEM for circumferences ranged from 0.5 cm to 1.9 cm. Unpaid volunteer measurers can, if carefully trained and supervised, provide acceptable blood pressure data and anthropometry in large population surveys. PMID- 11927201 TI - Cardiovascular risk factors and diseases precede oral hypoglycaemic therapy in patients with type 2 diabetes mellitus. AB - Although patients with type 2 diabetes mellitus and cardiovascular disease share common risk factors, the link between these diseases remains largely unexplained. In this case-control study, the earlier use of cardiovascular drugs (before the diagnosis of diabetes) was investigated among cases with type 2 diabetes mellitus and controls without diabetes. Using the PHARMO database, we identified 4,864 patients who were prescribed oral hypoglycaemic agent (OHA) therapy between 1985 1998 in the Netherlands. For each case, two controls matched on age, sex and pharmacy were randomly selected. Controls had not received insulins or OHA therapy. There were 2,656 (55.0%) cases compared with 2,727 (28.1%) controls who used cardiovascular drugs at the start of OHA therapy. Cases had a 3.5-fold increased risk of cardiovascular drug use (OR(95% CI) = 3.5 [3.2-3.8]) compared to controls. Differences in cardiovascular drug use were noted as early as 7 years before the start of OHA therapy, distinguishing cases from controls. Our finding that patients with type 2 diabetes mellitus were more likely to receive treatment for cardiovascular disease several years before they start diabetes therapy supports the hypothesis of a common underlying mechanism of these two disorders and stresses the importance of the pre-diabetic state. PMID- 11927202 TI - Assessing efficacy of chronic therapy when disease is characterized by frequent relapses: the example of schizophrenia. AB - In assessing the efficacy of therapy for schizophrenia, the effect of medication on relapse needs to be distinguished from the influence of relapse on medication. Typically, effective medication prevents relapse, but relapse generally induces medication. Conventional analyses using either treatment or disease as outcome do not separate these two effects. We propose an alternate approach that uses both treatment and schizophrenic relapse as random variables. Data from 58 schizophrenic patients, with up to 60 consecutive monthly determinations of antipsychotic medication and schizophrenic events, were analyzed using a bivariate transition model with random effects. This analysis revealed that the risk of current schizophrenic relapse is reduced by continuous medication (previous month and current month) but not necessarily by discontinuous medication. PMID- 11927203 TI - Incidence of chronic atrial fibrillation in general practice and its treatment pattern. AB - The object of this article was to estimate the incidence rate of chronic atrial fibrillation (AF) in a general practice setting, to identify factors predisposing to its occurrence, and to describe treatment patterns in the year following the diagnosis. The method used was a population-based cohort study using the General Practice Research Database (GPRD) in the UK. We identified patients aged 40-89 years with a first ever recorded diagnosis of AF. The diagnosis was validated through a questionnaire sent to the general practitioners. A nested case-control analysis was performed to assess risk factors for AF using 1,035 confirmed incident cases of chronic AF and a random sample of 5,000 controls from the original source population. The incidence rate of chronic AF was 1.7 per 1,000 person-years, and increased markedly with age. The age adjusted rate ratio among males was 1.4 (95% CI 1.2-1.6). The major risk factors were age, high BMI, excessive alcohol consumption, and prior cardiovascular comorbidity, in particular, valvular heart disease and heart failure. Digoxin was used in close to 70% of the patients, and close to 15% did not receive any antiarrhythmic treatment. Close to 40% did not receive either warfarin or aspirin in the 3 months period after the diagnosis. Among the potential candidates for anticoagulation only 22% of those aged 70 years or older were prescribed warfarin in comparison to 49% among patients aged 40-69 years. Chronic AF is a disease of the elderly, with women presenting a lower incidence rate than men specially in young age. Age, weight, excessive alcohol consumption, and cardiovascular morbidity were the main independent risk factors for AF. Less than half of patients with chronic AF and no contraindications for anticoagulation received warfarin within the first trimester after the diagnosis. PMID- 11927204 TI - Self-rated health, chronic diseases, and symptoms among middle-aged and elderly men and women. AB - The objective was to study the association between chronic diseases, symptoms, and poor self-rated health among men and women and in different age groups, and to assess the contribution of chronic diseases and symptoms to the burden of poor self-rated health in the general population. Self-rated health and self-reported diseases and symptoms were investigated in a population sample of 6,061 men and women aged 35-79 years in Varmland County in Sweden. Odds ratios (OR) and population attributable risks (PAR) were calculated to quantify the contribution of chronic diseases and symptoms to poor self-rated health. Depression, neurological disease, rheumatoid arthritis, and tiredness/weakness had the largest contributions to poor self-rated health in individuals. Among the elderly (65-79 years), neurological disease and cancer had the largest contribution to self-rated health in men, and renal disease, rheumatoid arthritis, and cancer in women. Among the middle-aged (35-64 years), depression and tiredness/weakness were also important, especially in women. From a population perspective, tiredness/weakness explained the largest part of poor self-rated health due to its high prevalence in the population. Depression and musculoskeletal pains were also more important than other chronic diseases and symptoms at the population level. Even though many chronic diseases (such as neurological disease, rheumatoid arthritis, and cancer) are strongly associated with poor self-rated health in the individual, common symptoms (such as tiredness/weakness and musculoskeletal pains) as well as depression contribute more to the total burden of poor self-rated health in the population. More preventive measures should therefore be directed against these conditions, especially when they are not consequences of other diseases. PMID- 11927205 TI - Validation of case-mix measures derived from self-reports of diagnoses and health. AB - Self-reported chronic diseases and health status are associated with resource use. However, few data exist regarding their ability to predict mortality or hospitalizations. We sought to determine whether self-reported chronic medical conditions and the SF-36 could be used individually or in combination to assess co-morbidity in the outpatient setting. The study was designed as a prospective cohort study. Patients were enrolled in the primary care clinics at seven Veterans Affairs (VA) medical centers participating in the Ambulatory Care Quality Improvement Project (ACQUIP). 10,947 patients, > or = 50 years of age, enrolled in general internal medicine clinics who returned both a baseline health inventory checklist and the baseline SF-36 who were followed for a mean of 722.5 (+/-84.3) days. The primary outcome was all-cause mortality, with a secondary outcome of hospitalization within the VA system. Using a Cox proportional hazards model in a development set of 5,469 patients, a co-morbidity index [Seattle Index of Co-morbidity (SIC)] was constructed using information about age, smoking status and seven of 25 self-reported medical conditions that were associated with increased mortality. In the validation set of 5,478 patients, the SIC was predictive of both mortality and hospitalizations within the VA system. A separate model was constructed in which only age and the PCS and MCS scores of the SF-36 were entered to predict mortality. The SF-36 component scores and the SIC had comparable discriminatory ability (AUC for discrimination of death within 2 y 0.71 for both models). When combined, the SIC and SF-36 together had improved discrimination for mortality (AUC = 0.74, p-value for difference in AUC < 0.005). A new outpatient co-morbidity score developed using self-identified chronic medical conditions on a baseline health inventory checklist was predictive of 2-y mortality and hospitalization within the VA system in general internal medicine patients. PMID- 11927206 TI - The effect of patient selection on comorbidity-adjusted operative mortality risk. Implications for outcomes studies of surgical procedures. AB - Consumers of outcomes research may assume that risk-adjustment procedures based on patients' comorbid conditions will control for baseline prognostic differences between comparison groups, so that differences in risk-adjusted outcomes represent effects other than those due to differences in comorbidity severity. However, surgeons may differ in their threshold to operate on patients with different intensities of the same comorbidity, which may not be accounted for using commonly employed risk-adjustment methods. We developed a model to explore the effect that selection based on comorbidity severity could have on estimates of the risk-adjusted relative risk (RR) of operative death. Larger effects on the apparent RR of operative death were observed when both the proportion of patients in the high-risk ("selected") stratum and the relative increase in the risk of death due to being in the high-risk stratum were large. Biased estimates of the risk-adjusted RR of operative death will be observed if surgeons differentially select patients based on comorbidity severity and if differences in comorbidity severity are not captured by the risk-adjustment methodology. PMID- 11927207 TI - Method of correction to assess the number of hospitalized incident breast cancer cases based on claims databases. AB - Estimations of the number of hospitalized incident cancer cases show biases when claims databases are used. This is due to false reports of incident cancer because of a lack of specificity, and because of unrecorded cancers resulting from a lack of sensitivity. We present a statistical method to provide corrected estimations. This method is based on a two-phase study design using an external data set for sensitivity and specificity estimates. Inaccuracy of the corrected number of hospitalized incident cancer cases was assessed by a credibility interval determined by a Bayesian approach using a Monte Carlo method. Based on the population hospitalized in a large group of French University hospitals, 334 women were identified in the French claims database as having potential incident cases of breast cancer in 1997. According to our method, the corrected number was 565 (550-580). In absence of hospital-based cancer registries, our approach provides estimates and credibility intervals, and has many potential applications in defining hospital policies with its applicability to other diseases. PMID- 11927208 TI - Estimating energy expenditure from the Minnesota Leisure Time Physical Activity and Tecumseh Occupational Activity questionnaires - a doubly labeled water validation. AB - An accurate, simple method for assessing energy expenditure in individuals and in free-living populations continues to be elusive. To compare estimates of energy expenditure (EE) from a combination of two previously validated physical activity questionnaires: Tecumseh Occupational (EE(TEC)) and a 4-wk history version of the Minnesota Leisure Time Physical Activity that included household activities (EE(MNLTPA)) and EE from sleep (EE(SLEEP)), to EE obtained from doubly labeled water (EE(DLW)). We studied free-living males (n = 24) eating a controlled diet designed to maintain body weight and determined EE from doubly labeled water (DLW) during 14 days and EE from physical activity instruments used in epidemiological studies (EE(TEC) and EE(MNLTPA)). There was excellent agreement between EE(DLW) (mean +/- SEM, 13.55 +/- 0.38 MJ/d) and EE(TEC) + EE(MNLTPA) + EE(SLEEP) (EE(TOTAL1)) (13.79 +/- 0.89 MJ/d) with a difference of only 1.0% +/- 5.4%. When the EE from watching TV, reading, and childcare activities was added the total EE (EE(TOTAL2)) (14.87 +/- 0.90 MJ/D) overestimated EE(DLW) by 8.9% +/- 5.4%. Both of these estimates of EE had significant regressions against EE(DLW) (EE(TEC) + EE(MNLTPA) + EE(SLEEP), R(2) = 0.38, P < 0.001; EE(TOTAL2), R(2) = 0.39, P < 0.001). Men whose occupations involved significant intermittent moderate activity had the largest disagreement between EE(DLW) and estimates from the questionnaires. This investigation demonstrates that a combination of previously validated physical activity questionnaires can be used to accurately determine the mean energy expenditure of a population of employed males. PMID- 11927209 TI - Comparing performance of multinomial logistic regression and discriminant analysis for monitoring access to care for acute myocardial infarction. AB - One way to monitor patient access to emergent health care services is to use patient characteristics to predict arrival time at the hospital after onset of symptoms. This predicted arrival time can then be compared with actual arrival time to allow monitoring of access to services. Predicted arrival time could also be used to estimate potential effects of changes in health care service availability, such as closure of an emergency department or an acute care hospital. Our goal was to determine the best statistical method for prediction of arrival intervals for patients with acute myocardial infarction (AMI) symptoms. We compared the performance of multinomial logistic regression (MLR) and discriminant analysis (DA) models. Models for MLR and DA were developed using a dataset of 3,566 male veterans hospitalized with AMI in 81 VA Medical Centers in 1994-1995 throughout the United States. The dataset was randomly divided into a training set (n = 1,846) and a test set (n = 1,720). Arrival times were grouped into three intervals on the basis of treatment considerations: <6 hours, 6-12 hours, and >12 hours. One model for MLR and two models for DA were developed using the training dataset. One DA model had equal prior probabilities, and one DA model had proportional prior probabilities. Predictive performance of the models was compared using the test (n = 1,720) dataset. Using the test dataset, the proportions of patients in the three arrival time groups were 60.9% for <6 hours, 10.3% for 6-12 hours, and 28.8% for >12 hours after symptom onset. Whereas the overall predictive performance by MLR and DA with proportional priors was higher, the DA models with equal priors performed much better in the smaller groups. Correct classifications were 62.6% by MLR, 62.4% by DA using proportional prior probabilities, and 48.1% using equal prior probabilities of the groups. The misclassifications by MLR for the three groups were 9.5%, 100.0%, 74.2% for each time interval, respectively. Misclassifications by DA models were 9.8%, 100.0%, and 74.4% for the model with proportional priors and 47.6%, 79.5%, and 51.0% for the model with equal priors. The choice of MLR or DA with proportional priors, or DA with equal priors for monitoring time intervals of predicted hospital arrival time for a population should depend on the consequences of misclassification errors. PMID- 11927210 TI - Citation bias of hepato-biliary randomized clinical trials. AB - The objective of this study was to assess whether trials with a positive (i.e., statistically significant) outcome are cited more often than negative trials. We reviewed 530 randomized clinical trials on hepato-biliary diseases published in 11 English-language journals indexed in MEDLINE from 1985-1996. From each trial, we extracted the statistical significance of the primary study outcome (positive or negative), the disease area, and methodological quality (randomization and double blinding). The number of citations during two calendar years after publication was obtained from Science Citation Index. There was a significant positive association between a statistically significant study outcome and the citation frequency (beta, 0.55, 95% confidence interval, 0.39-0.72). The disease area and adequate generation of the allocation sequence were also significant predictors of the citation frequency. We concluded that positive trials are cited significantly more often than negative trials. The association was not explained by disease area or methodological quality. PMID- 11927211 TI - Lifestyle and diet as risk factors for overanticoagulation. AB - The risk of hemorrhage when using coumarin anticoagulants sharply increases when the International Normalized Ratio (INR) is > or =6.0. We performed a case control study among outpatients of an anticoagulation clinic to identify sociodemographic-, lifestyle-, and dietary factors related to overanticoagulation. Three hundred cases with an INR > or =6.0 were compared with 302 randomly selected matched controls with an INR within the target zone. Age, sex, and level of education were not associated with overanticoagulation. Body mass index was negatively related to overanticoagulation, a beneath-average level of physical activity was positively related to overanticoagulation and never smokers were more likely to have an INR > or =6.0 compared with smokers. Habitual alcohol consumption, even heavy drinking, was not related to overanticoagulation. However, a recent decrease of alcohol intake increased the risk of an INR > or =6.0. In addition, weight loss and a vacation were risk factors for overanticoagulation. Dietary factors were not associated with overanticoaguation. If risk factors can not be avoided, increased monitoring of INR values could prevent overanticoagulation and potential bleeding complications. PMID- 11927213 TI - Modeling bivariate ordinal contingency tables arising in studies of interobserver variation, with application to cervical screening. PMID- 11927212 TI - Did Weir Mitchell anticipate important concepts in ambulatory care and clinical epidemiology? AB - During the decade from 1977 to 1986, four models of care pertaining to ambulatory medicine and certain aspects of clinical epidemiology were proposed. All were concerned with the frequently heard accusation that medicine was becoming dehumanized by being overly dependent on powerful new technologies. Some of the authors went so far as to suggest that the view, according to which medical science must restrict itself to "hard" data of the kind provided by the serum multichannel analyzer, should be considered outdated and, in fact, unscientific. Their plea was to develop a science of the clinical encounter that would shift the emphasis from explication to prediction and management, the latter term being virtually synonymous with decision making. For this change to come about, they wrote, it would be necessary to collect "soft" data on such subjects as family relationships, psychic traits and states, perceptions of life quality, patient expectations and many others. We believe that some of these subjects as well as the models themselves were anticipated in the writings, both medical and fictional, of Weir Mitchell, nearly a century earlier. This paper, after presenting a brief overview of the career of a colorful and commanding figure from the annals of American medicine, will seek to illustrate his extraordinary farsightedness as a practitioner of primary care and his relevance for some aspects of clinical epidemiology. Because the attempts to link his ideas to modern concepts are ours, we accept the possibility that, here and there, we may have read things into his writings that he did not intend. PMID- 11927214 TI - Choosing effect measures for epidemiologic data. PMID- 11927216 TI - Shear stress and vascular remodeling: study of cardiac allograft coronary artery disease as a model of diffuse atherosclerosis. AB - BACKGROUND: The assessment of remodeling in diffuse atherosclerosis by intravascular ultrasound is hampered by the lack of an appropriate reference segment. Transplant coronary artery disease is an accepted model of diffuse atherosclerosis. Flow-dependent remodeling is regulated by shear stress. Thus, normal levels of shear stress at baseline flow reflect adequate regulation and provide a functional assessment of flow-dependent remodeling. METHODS: The approach was evaluated in 91 patients with transplant coronary artery disease and in 9 non-transplanted controls and 16 control transplant recipients. Quantitative coronary angiography, intracoronary ultrasound and intracoronary velocity studies were performed at baseline and after pharmacologic intervention. Calculated shear stress was compared between these groups and a sub-group with coronary angioplasty at follow-up (8 of 60 patients with control angiography after 23 +/- 8 months). The relation of shear stress to flow, diameter, flow/area ratio and endothelial function was analyzed. RESULTS: Normal shear stress was found in non transplanted controls and transplant recipients without coronary artery disease. Patients with coronary angioplasty at follow-up had elevated shear stress and enhanced endothelial dysfunction on the initial investigation. Shear stress was not correlated with flow (r = 0.062, non-significant), but with diameter (r = 0.654), flow/area ratio (r = 0.814) and endothelial dysfunction (r = 0.722). CONCLUSION: Calculated local shear stress appears to be useful for the assessment of the adequacy of flow-dependent macrovascular remodeling in diffuse atherosclerosis. Elevated blood flow/area ratio is a potential clinical marker of increased shear stress that reflects inadequate flow-dependent remodeling. PMID- 11927217 TI - Ex vivo blockade of endothelin-1 inhibits graft coronary artery disease in a rodent cardiac allograft model. AB - BACKGROUND: Graft coronary artery disease (GCAD) is characterized by vascular narrowing resulting from intimal hyperplasia. Endothelin (ET)-1, derived from the vascular endothelium and macrophages, stimulates vascular smooth muscle cells (SMCs), which leads to neointimal formation in donor graft coronary arteries. In this study, we hypothesized that antisense (AS) oligodeoxynucleotides (ODN) for preproendothelin-1 (ppET-1) delivered to rat cardiac allografts by means of hyperbaric pressure would reduce the incidence of GCAD. METHODS: PVG donor hearts were infused with ppET-1 AS ODN (80 micromol/liter), sense ODN, scrambled ODN or saline alone and incubated in a pressure chamber at 75 psi or ambient pressure for 45 minutes. Cardiac allografts were heterotopically transplanted into ACI rats treated with cyclosporine (7.5 mg/kg, Days 0 to 9). Allografts were procured at post-operative days (POD) 7 or 90. Reverse transcription-polymerase chain reaction (RT-PCR) assay for ET-1 mRNA and ET01 immunohistochemistry (IHC) were performed at PODs 7 and 90. Elastic staining and IHC with anti-macrophage and alpha-SMC actin antibodies were performed to assess GCAD at POD 90. RESULTS: Treatment with AS ODN and pressure significantly reduced ET-1 mRNA and protein expression. A significant reduction in GCAD was achieved with inhibition of ET-1 and was associated with attenuation of macrophages and SMCs in the neointima. CONCLUSION: Peri-operative ex vivo inhibition of ET-1 expression results in a reduction of GCAD. This highly targeted therapy may be a clinically viable strategy for the prevention of ET-1-induced GCAD following cardiac transplantation. PMID- 11927218 TI - Cyclosporine does not enhance the development of accelerated coronary artery disease: experimental study in a rat cardiac transplant model. AB - BACKGROUND: Among other factors, cyclosporine (CsA) is linked with the development of accelerated coronary artery disease (ACAD) after transplantation. The objective of this study was to assess the influence of different CsA regimens on ACAD after rat heart transplantation. METHODS: After heterotopic cardiac transplantation (Lewis to Fisher), animals were treated with 3 or 12 mg/kg per day of CsA, administered subcutaneously. The control group received no therapy. CsA blood levels were determined every 10 days. Twenty and 80 days after grafting, the incidence of ACAD was determined and the extent of ACAD was assessed as mean vessel occlusion (mvo). RESULTS: In the 12-mg group, CsA levels were nearly 10-fold higher than in the 3-mg group. Only in the 12-mg CsA group was the incidence of ACAD significantly reduced at Days 20 and 80 when compared with controls. Continuous therapy with 3 mg and 12 mg of CsA significantly reduced the mvo at Days 20 and 80 when compared with control animals (p <.05). However, comparing the two dosages, there were no significant differences. When the 20-day-limited course of CsA application was used we did not observe significant differences in mvo at Day 80 upon comparison of 3-mg and 12-mg CsA treatment versus untreated animals. CONCLUSIONS: Despite the excessive increase in CsA blood levels we observed neither a further reduction nor an increase of ACAD in the high-dose group compared with the low-dose group. Therefore, CsA did not enhance the development of chronic rejection in this experiment. PMID- 11927219 TI - Efficacy of tacrolimus rescue therapy in refractory acute rejection after lung transplantation. AB - BACKGROUND: Encouraging results in transplantation of other solid organs led to investigation of the use of tacrolimus in lung transplantation as a salvage immunosuppressant in persistent acute rejection. METHODS: The incidence and severity of acute rejection and the number of steroid pulses were analyzed in 20 lung recipients who were converted from a cyclosporine- to a tacrolimus-based immunosuppressive regimen because of refractory biopsy-proven acute rejection. RESULTS: Tacrolimus was started 12.0 +/- 13.0 months after transplantation, and the mean follow-up was 25.0 +/- 13.7 months. After shifting to tacrolimus, a significant decline was observed in both the number of acute rejections per patient (3.0 +/- 1.56 to 0.85 +/- 1.14, p < 0.0001), and the incidence of acute rejection per 100 patient-days (1.52 +/- 0.99 to 0.14 +/- 0.21, p < 0.0001). Furthermore, the average histologic grade of rejection decreased from 1.9 +/- 0.8 to 0.4 +/- 0.5 (p < 0.0001). Methylprednisolone pulses similarly decreased from 1.9 +/- 1.3/patient to 0.3 +/- 0.7/patient (p < 0.0001). During cyclosporine immunosuppression, the mean forced expiratory volume in 1 second decreased to 84.4% +/- 13.3% of individual best value. The average lung function parameters were stable 3 months after the change of medication, and then began to improve. After an average follow-up of 36.5 +/- 19.2 months, 2 patients have developed bronchiolitis obliterans syndrome (one has Stage 1 and one has Stage 3). CONCLUSION: Conversion to a tacrolimus-based immunosuppressive regimen for refractory acute lung rejection is associated with reduced incidence and severity of acute rejection episodes, steroid sparing, and stabilization or improvement of pulmonary function. PMID- 11927220 TI - HMG-CoA reductase inhibitor cerivastatin prolonged rat cardiac allograft survival by blocking intercellular signals. AB - BACKGROUND: The development of atherosclerotic cardiovascular complications caused by hyperlipidemia is a common and serious problem for long-term survivors of organ transplantation. However, adhesion molecules such as intercellular adhesion molecule (ICAM)-1 and lymphocyte function-associated antigen (LFA)-1 are involved in allograft rejection, possibly by providing costimulatory signals. 3 Hydroxy-3-methylglutaryl coenzyme A (HMG-CoA) reductase inhibitor cerivastatin has been shown to suppress ICAM-1 expression in acute inflammatory responses. METHODS: In this study, we evaluated the immunosuppressive effects of cerivastatin in rat cardiac allografts. The hearts of Fischer rats were transplanted heterotopically into Lewis rats. Cerivastatin (2 mg/kg) was administrated intraperitoneally to recipients for 7 consecutive days from the day before transplantation. RESULTS: Graft survival in the cerivastatin-treated group (n = 8) was significantly longer than in controls (n = 10) (24.6 +/- 2.2 days vs 10.2 +/- 1.3 days, p < 0.05). Mixed lymphocyte reaction (MLR) showed that on Day 8 after grafting, the proliferative response of alloreactive T cells against F344 alloantigen in cerivastatin-treated rats was significantly more suppressed than in Lewis rats. The Interleukin-2 concentration of supernatant in MLR cultures in the cerivastatin-treated group was lower than in the control group. Immunohistochemical analysis showed that the percentage of CD4-positive cells to infiltrating mononuclear cells was less prominent in the cerivastatin-treated group (9.8% +/- 2.2%) than in the control group (20.9% +/- 3.2%). CONCLUSIONS: The HMG-CoA reductase inhibitor cerivastatin effectively suppressed acute graft rejection, possibly by blocking intercellular signals via ICAM/LFA-1, and cerivastatin may be a candidate for treating patients with hyperlipidemia who undergo organ transplantation. PMID- 11927221 TI - Partial loading of the left ventricle during mechanical assist device support is associated with improved myocardial function, blood flow and metabolism and increased exercise capacity. AB - BACKGROUND: Myocardial recovery has been observed after placement of left ventricular assist devices in some patients awaiting cardiac transplantation. Left ventricular assist devices provide profound volume and pressure unloading while restoring systemic blood flow. However, the optimal degree of left ventricular unloading during left ventricular assist device support is unknown. The purpose of this study was to assess the effect of the degree of left ventricular decompression, during left ventricular assist device support, on myocardial function and exercise capacity. METHODS: Twenty patients with a left ventricular assist device performed cardiopulmonary exercise testing with simultaneous echocardiographic and hemodynamic measurements during full, and then partial, device support. Eleven patients underwent positron emission tomographic scanning for measurement of myocardial blood flow and oxygen consumption. RESULTS: Patients were divided into two groups based on the degree of left ventricular decompression as assessed by echocardiographic measurements. Patients with partially decompressed ventricles (n = 13, LVEDD = 4.8 +/- 1.0 cm) as compared to those with fully decompressed ventricles (n = 10, LVEDD = 3.0 +/- 0.3 cm) demonstrated significant improvements in: peak oxygen consumption (16.8 +/- 4.3 versus 12.8 +/- 3.0 ml/kg.min), native left ventricular index during exercise (2.5 +/- 1.4 versus 0.8 +/- 0.8 liters/min.m(2)); ability to exercise with the device weaned (10 of 13 versus 1 of 10 patients); resting myocardial blood flow (0.55 +/- 0.11 versus 0.21 +/- 0.13 ml/g.min); and myocardial oxygen consumption (0.04 +/- 0.01 versus 0.02 +/- 0.001 min-1) (all p < 0.05). CONCLUSIONS: These results suggest that partial loading of the left ventricle during left ventricular assist device support may be beneficial. Further study of optimal ventricular decompression during device support is needed, as this may be important in improving myocardial recovery. PMID- 11927222 TI - Cardiac transplantation for pediatric restrictive cardiomyopathy: presentation, evaluation, and short-term outcome. AB - BACKGROUND: Because of the poor prognosis of pediatric restrictive cardiomyopathy, transplantation has been proposed as the treatment of choice for this disease. METHODS: We reviewed our experience with the presentation, evaluation, and short-term outcome in 8 pediatric patients with restrictive cardiomyopathy referred for transplantation. Potential reversibility of elevation in pulmonary vascular resistance was tested before transplantation with nitroprusside and nitric oxide, with follow-up cardiac catheterization performed 6 to 12 months after transplantation. RESULTS: The mean age of diagnosis of restrictive cardiomyopathy was 6.3 years and the mean interval from diagnosis to referral for transplantation was 3.6 years. Elevation of pulmonary vascular resistance was common and tended to progress with longer follow-up. Three of the 8 patients had pulmonary vascular resistance indices greater than 10 Woods unit/m(2) and transpulmonary gradients greater than 20 mm Hg. The administration of nitroprusside and nitric oxide reversed elevated pulmonary resistance and transpulmonary gradients in all patients. Nitric oxide successfully reversed pulmonary vascular resistance in patients unresponsive to nitroprusside. All patients underwent successful transplantation and follow-up catheterization revealed normal pulmonary hemodynamics in each patient. CONCLUSIONS: Pediatric restrictive cardiomyopathy can be associated with marked elevation in the pulmonary vascular resistance, which may contribute to the poor prognosis in these patients and potentially make cardiac transplantation problematic. Orthotopic cardiac transplantation can be successfully performed in patients who demonstrate reversibility of pulmonary vascular resistance. Nitric oxide appears to be the best agent to demonstrate reversibility of pulmonary resistance in these patients. PMID- 11927224 TI - Results of an artificial-lung survey to lung transplant program directors. AB - BACKGROUND: Lung transplantation is the only treatment for patients with end stage lung disease. However, the scarcity of donor organs illustrates the need for alternatives. Recent success in the use of ventricular assistance has stimulated research in technology designed as a bridge to lung transplantation. Some laboratories have demonstrated significant advances in the development of artificial lungs, and clinical applications are on the horizon. In preparation, we sought to gather information from the lung transplant community regarding issues related to testing and potential trials of artificial lungs. METHODS: We constructed a survey and distributed it to lung transplant program directors recognized by the United Network for Organ Sharing. Topics included required animal studies, preferred designs, logistics of clinical trials, and patient diagnoses most appropriate for such a trial. RESULTS: The 31 programs responding to the survey performed 72% of all lung transplants in the United States in 1999. Ninety-seven percent supported a Phase I trial using an artificial lung as a bridge to lung transplantation. Additionally, 58% specifically supported a trial in which organ allocation would be prioritized to enrolled patients. Idiopathic pulmonary fibrosis was the diagnosis thought most appropriate for inclusion in initial clinical trials. CONCLUSIONS: Widespread support exists for the development and use of an artificial lung as a bridge to lung transplantation. Information from transplant centers regarding device design and application can influence laboratories developing artificial lungs, and such communication will be essential as this technology progresses from the bench-top to the bedside. PMID- 11927223 TI - Valacyclovir prevention of cytomegalovirus reactivation after heart transplantation: a randomized trial. AB - BACKGROUND: Cytomegalovirus (CMV) is a major cause of serious morbidity following solid organ transplantation via both direct and indirect mechanisms. The aim of this study was to investigate the efficacy and safety of valacyclovir prophylaxis in heart transplant recipients. METHODS: Twenty-seven CMV seropositive adults due to receive a heart transplant were included in a single-center, randomized, double-blind study. Patients were randomized to receive either oral valacyclovir 2000 mg or oral acyclovir 200 mg four times daily starting within 3 days of heart transplant and continuing for 90 days. The primary outcome measure was time to development of CMV antigenemia assessed for 6 months after surgery. Other measures were time to asymptomatic CMV infection, symptomatic CMV infections, and end-organ CMV disease. Patients were monitored for other herpes infections, other opportunistic infections, and acute graft rejection. Safety was assessed by evaluating changes in hematology and clinical chemistry parameters and by the occurrence of adverse events. RESULTS: The median time to CMV antigenemia was 19 days for the acyclovir group compared with 119 days for the valacyclovir group (hazard ratio 0.42; 95% CI, 0.18-0.99; p = 0.049). Similar delays of approximately 100 days were found for CMV infection, symptomatic CMV infection, and CMV disease. There was also a trend for delayed acute rejection, and fewer opportunistic or other herpesvirus infections occurred in the valacyclovir group. Valacyclovir was well tolerated in the study population. CONCLUSION: Oral valacyclovir is a safe and effective mode of prophylaxis of CMV after heart transplantation. PMID- 11927225 TI - Histopathology of cardiac xenograft rejection in the pig-to-baboon model. AB - BACKGROUND: The use of pig organs transgenic for human decay accelerating factor (hDAF) has largely overcome the problems of hyperacute rejection. With improved immunosuppressive protocols, life supporting grafts are showing greater survival times bringing the possibility of clinical xenotransplantation closer. Examination of the histopathology of the rejection process provides insight into the underlying mechanism and may suggest ways in which new immunosuppressive strategies should be directed. METHODS: 44 baboons (Papio anubis) underwent heart transplants of which 39 were from transgenic donors. The transplanted organs were examined histologically and stained for evidence of immunoglobulin and complement deposition as well as cellular infiltrates. RESULTS: In the transgenic animals survival times were 2 to 99 days (mean 23.5) and the heterotopic group and 1 to 39 days (mean 11.7) in the orthotopic group. There were 3 cases of hyperacute rejection between the 2 groups. Rejected organs showed areas of old and recent myocardial infarction associated with vascular thrombosis. There was widespread deposition within vessels of immunoglobulins IgM and IgG together with complement fractions C3 and C5b to 9 in those organs that were rejected. The amount of complement positive in the longer surviving organs was less than those rejecting early. Cellular infiltate was predominantly macrophage with some later appearing T or natural killer cells. CONCLUSIONS: The histopathological changes support the importance of immunoglobulin and complement in delayed xenograft or acute vascular rejection. With time there is an increase in cellular infiltrate predominantly macrophages and these findings suggest an increasingly important role for the cells and the rejection process. The presence of areas of infarction and underlying vascular thrombosis is in keeping with endothelial activation and the establishment of procoagulant phenotype which may be due to immunoglobulin, complement, secreted cytokines and direct cellular effects. PMID- 11927226 TI - Removal of cartilage rings prevents graft stenosis in extended tracheal allotransplantation with omentopexy and immunosuppression: an experimental study. AB - BACKGROUND: One of the serious problems in longer size tracheal transplantation is severe stenosis of the graft, probably caused by an inadequate blood supply. We have previously reported that removal of some cartilage rings of the graft and omentopexy helps to provide sufficient blood flow to the graft mucosal tissue and results in satisfactory survival and non-significant graft stenosis in extended tracheal autotransplantation. However, it is unclear whether this method can be applied to extended tracheal allotransplantation that requires immunosuppression. In this report, we describe midterm results of extended tracheal allotransplantation with the technique. METHODS: Twenty-four adult mongrel dogs were used. In 18 dogs, a nine-cartilage-ring length of the trachea was allotransplanted when five cartilage rings of the graft were removed, leaving two rings intact at both ends of the graft for simple fixing to the recipient. Two artificial tracheal rings outside the graft and a stent inside the graft were used for maintaining the lumen width. Omentopexy was done for sufficient blood supply to the graft. FK 506 (0.1 mg/kg) was given on each day after the operation in Group A (n = 10), but was not given at all in Group B (n = 8). In Group C (n = 6), a nine-cartilage-ring length of the trachea, without removal of any cartilage ring, was transplanted into the recipient dog and covered with an omental pedicle flap. The same dose of FK 506 as that used in Group A dogs was given to Group C dogs. RESULTS: In Group A, 2 dogs died of graft stenosis within 9 weeks after surgery and 1 died of emaciation without tracheal stenosis. Seven dogs (70%) survived until time of killing. Among the 8 dogs in Group B, 6 died of graft stenosis within 9 weeks after surgery, with 1 dying of pneumonia and only 1 (13%) surviving for >1 year until killing. In Group C, all 6 dogs died of graft stenosis within 6 weeks after surgery. Survival at 16 weeks after surgery was 70% in Group A, 13% in Group B and 0% in Group C (p < 0.01, A vs B and C). No significant graft stenosis was found in 6 dogs and mild stenosis was found in 2 dogs at the time of death or killing in Group A (80%), whereas mild stenosis was found in only 2 dogs in Group B (25%) (p < 0.05). Mucosal blood flow of the graft in Group A was higher than that in Group C and was the same as that in Group B within 4 weeks after surgery; however, it remained unchanged to ultimately be higher than in Group B at 6 and 8 weeks after surgery. CONCLUSIONS: Removal of some cartilage rings, omentopexy and immunosuppression improved blood supply to the graft and resulted in good survival and non-significant tracheal stenosis in extended tracheal allotransplantation. PMID- 11927227 TI - Hormone therapy in men and risk of cardiac allograft rejection. AB - Hormone therapy with leuprolide acetate (Lupron) is frequently used to treat prostate cancer, a common malignancy expected to be diagnosed in >180,000 men in the United States this year. In addition, testosterone supplementation is frequently recommended for adult men with reduced libido or impotence and decreased serum levels of testosterone. Although these hormone therapies are generally considered benign, we describe 3 cases of acute cardiac rejection temporally associated with the use of these drugs in men who have undergone cardiac transplantation. To our knowledge, this is the first such description of this phenomenon in the literature. PMID- 11927228 TI - Becker muscular dystrophy-related cardiomyopathy: a favorable response to medical therapy. AB - Patients with Becker muscular dystrophy-related cardiomyopathy typically survive into their 30s, when they succumb to complications of cardiomyopathy or receive heart transplants. The cardiomyopathy is related to an abnormality in the protein dystrophin; no successful medical responses have been reported to date. We describe a patient with Becker muscular dystrophy-related cardiomyopathy in whom we noted improvement of cardiac status with carefully titrated medical therapy. Successful therapy suggests a trial of medical therapy may be beneficial when patients with Becker muscular dystrophy-related cardiomyopathy are evaluated for transplant. PMID- 11927229 TI - Acquired supravalvar aortic stenosis following heart transplantation: report of 2 cases. AB - We report 2 cases of acquired supravalvar aortic stenosis (SVAS) complicating orthotopic cardiac transplantation that contributed to the post-operative death of both patients. In the first patient the stenosis resulted from kinking of a too-long graft aortic component that was rendered into a fixed deformity by healed mediastinitis and constrictive pericarditis. Supra-aortic stenosis in the second patient occurred at a site of multiple aortic cannulations that was surrounded by dense fibrosis. Acquired SVAS contributed to the death of the first patient (who had graft arteriopathy) and was the proximate cause of death in the second patient. Specific echocardiography of the ascending aorta, which is not routinely performed, may detect SAVS. PMID- 11927230 TI - Disseminated Cladophialophora bantiana infection in a heart transplant recipient. AB - Cerebral phaeohyphomycosis caused by Cladophialophora bantiana, a dematiaceous fungus, is a rare disease. The majority of cases have been reported among immunocompetent patients; only 4 cases have been published that describe transplantation patients. The overall prognosis is poor. Surgical therapy in combination with chemotherapy with itraconazole is recommended. We report the case of a heart transplant recipient with cutaneous, cerebral, and lung manifestation of Cladophialophora bantiana who died despite surgical and systemic, high-dosage itraconazole treatment. PMID- 11927231 TI - Lung transplantation in a patient with arteriovenous malformations. AB - Multiple arteriovenous fistulas (AVF) is an infrequent pathology, in which lung transplant is controversial. We report a patient with hereditary hemorrhagic telangiectasia who underwent a single lung transplant with excellent evolution, and three years later (that is, at the time of this paper), she is still alive. PMID- 11927232 TI - Looking back to the future. Stress in adolescents: gender differences in relationship to autoimmune disease. PMID- 11927233 TI - School-based hepatitis B immunization program in adolescents. PMID- 11927234 TI - The association between impulsiveness and sexual risk behaviors in adolescent and young adult women. AB - Cross-sectional survey data from 462 female adolescents were analyzed to determine associations between impulsiveness and sexual risk behaviors and related outcomes. Higher impulsiveness was significantly associated with early age of first sexual intercourse, higher number of sexual partners, non-use of contraception and condoms, and prior Chlamydia infection. PMID- 11927235 TI - Adolescent boys and the muscular male body ideal. PMID- 11927236 TI - Steroid use among adolescents: findings from Project EAT. AB - PURPOSE: To explore the prevalence and correlates of using steroids for the purpose of gaining muscle among adolescent males and females. More specifically, the study objectives were to: (a) assess the prevalence of anabolic steroid use in a large population-based sample of adolescent males and females; and (b) identify personal, socio-environmental, and behavioral factors related to the health of adolescents that use anabolic steroids. METHODS: The study population included 4746 middle and high school students from St. Paul/Minneapolis public schools who completed surveys and anthropometric measurements as part of a population-based study of eating patterns and weight concerns among teenagers (Project EAT: Eating Among Teens). Steroid use and a range of socio environmental, personal, and behavioral factors hypothesized to be correlates of steroid use were assessed. Associations were examined separately among males and females through comparisons of means (unadjusted and adjusted for grade-level and race) for continuous variables, and percentages, and odds ratios for dichotomized variables. RESULTS: Steroid use was more common in males (5.4%, vs. 2.9% of females), non-Caucasians (especially Hmong, a subset of the Asian-American population), and in middle school students (as compared to high school). In males, steroid use was associated with poorer self-esteem and higher rates of depressed mood and attempted suicide, poorer knowledge and attitudes about health, greater participation in sports that emphasize weight and shape, greater parental concern about weight, and higher rates of disordered eating and substance use. Among females, steroid use was less consistent in its associations with other variables, although overall, a similar pattern of results emerged. CONCLUSIONS: Anabolic steroid use in adolescence is associated with poorer health related attitudes and behaviors and exposure to socio-environmental influences encouraging weight preoccupation. Attention needs to be directed toward youth who may be at increased risk for steroid use within clinical and community-based settings PMID- 11927238 TI - Health care use and charges for adolescents enrolled in a title XXI program. AB - PURPOSE: To examine the use patterns and charges for adolescents with special health care needs (ASHCN) and adolescents engaging in risky behaviors (ARB) and both (BOTH), compared with adolescents with no identified special health care or risky behavior diagnosis (ANIC) in a pool of adolescent enrollees eligible for Title XXI services. METHODS: Claims and encounter data were used to classify 11,459 who had been enrolled in Florida's Healthy Kids Program for 2 consecutive years into 4 groups: ASHCN (n = 1363); ARB (n = 1801); BOTH (n = 773); and ANIC (n = 7522). Monthly per capita inpatient, outpatient, emergency room, and total use and charges were calculated and compared across groups using the Wilcoxon rank sum test, descriptive statistics, and odds ratios. RESULTS: The data indicated that ASHCN, ARB, and BOTH consume significantly more health care services than ANIC and that their higher use and charges are attributable not only to condition-related costs but also to their higher use of non-condition related services as well. The data also revealed that adolescents with a special health care diagnosis (ASHCN and BOTH) had a higher percentage of costs owing to inpatient stays, whereas adolescents with a risky behavior diagnosis (ARB and BOTH) had a higher percentage of costs owing to emergency room usage. Finally, relevant to the total resources necessary to provide health care for adolescents, the data showed that 65% of the total annual costs of this program emanate from the 35% of adolescents who have special health care needs, engage in risky behaviors, or both. CONCLUSIONS: This study underscores two important points that can be used to ensure that adequate financing, provider network, and service delivery systems are developed to best meet the needs of the adolescent population: (a) the importance of considering the unique needs of adolescents in reimbursement rate setting strategies and (b) the importance of considering category of service utilization in weighting premiums for various groups. PMID- 11927237 TI - Parental correlates of physical activity in a racially/ethnically diverse adolescent sample. AB - PURPOSE: To evaluate relationships between parents' and adolescents' physical activity and television usage and whether these relationships differed among adolescents from different racial/ethnic backgrounds. METHODS: Adolescents and their parents were separately asked to report information about their eating, exercising, and weight-related behaviors. Among the 900 adolescents, 477 were girls and 423 were boys; 60% were in high school; 29% were white, 23% were black, 21% were Asian, 14% were Hispanic, and 13% were considered mixed or other race/ethnicity. RESULTS: Parents' reported encouragement was positively related to physical activity in white (r = 0.39; p <.001) and black boys (r = 0.26; p =.007), and girls (all race/ethnic groups combined: r = 0.15; p <.001). Parents' television time was positively related to television time in Hispanic boys (r = 0.40; p =.009) but negatively related to television time in black boys (r = 0.23; p =.036). Parents' concern about their own fitness was negatively related to television time in white girls (r = -0.19; p =.029) but positively related in black girls (r = 0.23; p =.030). CONCLUSION: This study found significant, although modest, relationships between parents' and adolescents' physical activity attitudes and behaviors. Many of these relationships differed by race/ethnicity. Results from the present and previous studies suggest that factors other than parents' behavior and support explain adolescents' physical activity behaviors. PMID- 11927239 TI - Reduced emergency department utilization associated with school-based clinic enrollment. AB - PURPOSE: To evaluate the change in emergency department utilization before and after enrollment in a school-based clinic (SBC). METHODS: This study was a retrospective cohort study comparing emergency department visit rate before and after SBC enrollment based on chart review. An initial cohort of 10- to 15-year olds who used a local emergency department was divided into two samples: those who had been enrolled in the SBC and those who had not. Emergency department charts were reviewed for visits that occurred during the initial year and two subsequent school years. The change in visit rate for each sample group was measured for each school year evaluated. RESULTS: The initial cohort consisted of 2852 youth, 344 of whom who lived in the high school attendance zone. Of these, the number of students enrolled in both the high school and the SBC was 43; the number enrolled in the high school but not in the clinic was 48. The emergency department visit rate decreased for both groups for each school year evaluated; however, this decrease was significant only for the SBC sample with a 41% to 57% decrease in emergency department utilization, 18% greater than the decrease in students who did not use the SBC. CONCLUSIONS: Accessible, prevention-oriented health care provided in a SBC can decrease the utilization of episodic health care in an emergency department. PMID- 11927240 TI - Risk factors for smoking among adolescents with asthma. AB - PURPOSE: To examine the prevalence of smoking among youth with asthma and explore potential risk factors for smoking among these youth, compared with youth without asthma. METHODS: In March 1997, a questionnaire assessing risk factors for smoking was administered to students attending public high school in Memphis, Tennessee. The sample (n = 3234) ranged from 15 to 18 years of age, was predominately African-American (76.7%), and 56.7% female. Based on a single self reported item, 15.7% of youth reported having asthma. Logistic regression analyses assessed asthma status differences in current smoking and the association between risk factors and smoking. RESULTS: Nearly 20% of youth reported current smoking. Adolescents with asthma (current or past) were nearly 1.5 times more likely to smoke than peers without asthma. The association between risk factors and current cigarette smoking was equivalent for youth regardless of whether they had asthma and did not differ for those youth with current vs. past asthma. CONCLUSIONS: Results suggest that adolescents with or without asthma smoke for similar reasons. Research is needed to elucidate whether there are unique risk factors, not examined here, for smoking among adolescents with asthma. Smoking prevention and cessation programs are needed for youth with asthma. PMID- 11927241 TI - Sex behavior among high school students in Sweden: improvement in contraceptive use over time. AB - PURPOSE: To investigate sexual behavior and attitudes toward sexuality among first-year high school students in Sweden in 1999 and to compare the sexual behavior with that reported from similar studies in 1979 and 1989. METHODS: In Spring 1999, a questionnaire with 47 questions mainly of multiple-choice nature was administered to a random sample of 20 classes in senior high school in two medium-sized cities in Sweden. The median age of the students was 16 years. Differences in sexual behavior and attitudes were evaluated with the Chi-square and Student's t-test. RESULTS: Almost half of the students (46%) had had intercourse, showing that the age of coitarche had not decreased during the past two decades. The median age at intercourse of those who had had intercourse was 15 years in both genders. More students in practical (vocational-technical) programs (60%) than in theoretical (college preparatory) ones (37%) had had sexual intercourse. More teenagers in theoretical (90%) than in practical programs (64%) had used contraceptives at their first intercourse, an increase from 65% in 1979 to 76% in 1999. One-fourth of the teenagers were under the influence of alcohol at the first intercourse, a decrease, compared with 1979. CONCLUSIONS: Contraceptive use at coitarche has increased and alcohol use decreased over the 20-year-period. The tendency noted 10 years ago that students attending practical programs put themselves at more health risk than students in theoretical programs continues. This fact should be considered when planning for counseling and sex education. PMID- 11927253 TI - DNA-cation interactions: quo vadis? AB - The interactions between double helical DNA and cations, specifically mono- and divalent metal ions, have recently received increased attention. Molecular dynamics simulations, solution NMR, and X-ray crystallography have all shed light on the coordination of ions in the major and minor grooves of DNA. Metal ion interactions may play key roles in the control of DNA conformation and topology, but despite progress in locating the ions and determining their precise binding modes, it remains difficult to figure out just how important ions really are. What have we learned and what remains to be done? PMID- 11927254 TI - Engineered urdamycin glycosyltransferases are broadened and altered in substrate specificity. AB - Combinatorial biosynthesis is a promising technique used to provide modified natural products for drug development. To enzymatically bridge the gap between what is possible in aglycon biosynthesis and sugar derivatization, glycosyltransferases are the tools of choice. To overcome limitations set by their intrinsic specificities, we have genetically engineered the protein regions governing nucleotide sugar and acceptor substrate specificities of two urdamycin deoxysugar glycosyltransferases, UrdGT1b and UrdGT1c. Targeted amino acid exchanges reduced the number of amino acids potentially dictating substrate specificity to ten. Subsequently, a gene library was created such that only codons of these ten amino acids from both parental genes were independently combined. Library members displayed parental and/or a novel specificity, with the latter being responsible for the biosynthesis of urdamycin P that carries a branched saccharide side chain hitherto unknown for urdamycins. PMID- 11927255 TI - RNA-catalyzed RNA ligation on an external RNA template. AB - Variants of the hc ligase ribozyme, which catalyzes ligation of the 3' end of an RNA substrate to the 5' end of the ribozyme, were utilized to evolve a ribozyme that catalyzes ligation reactions on an external RNA template. The evolved ribozyme catalyzes the joining of an oligonucleotide 3'-hydroxyl to the 5' triphosphate of an RNA hairpin molecule. The ribozyme can also utilize various substrate sequences, demonstrating a largely sequence-independent mechanism for substrate recognition. The ribozyme also carries out the ligation of two oligonucleotides that are bound at adjacent positions on a complementary template. Finally, it catalyzes addition of mononucleoside 5'-triphosphates onto the 3' end of an oligonucleotide primer in a template-dependent manner. The development of ribozymes that catalyze polymerase-type reactions contributes to the notion that an RNA world could have existed during the early history of life on Earth. PMID- 11927256 TI - Insight into binding of calyculin A to protein phosphatase 1: isolation of hemicalyculin a and chemical transformation of calyculin A. AB - Calyculin A isolated from the marine sponge Discodermia calyx is a potent inhibitor of protein phosphatases 1 and 2A. We attempted to elucidate its mode of binding to the enzymes by examining the activity of natural and chemically transformed derivatives. Ten natural derivatives including a new compound, hemicalyculin A, were provided. The structure of hemicalyculin A, which comprises the southern hemisphere of calyculin A, was firmly established by chemical methods. Six compounds were prepared by selective modifications of functional groups in calyculin A. The enzyme inhibitory activity of these compounds indicated that 17-phosphate, 13-hydroxyl, and the hydrophobic tetraene moieties were all necessary for binding to the enzymes. The derivatives lacking the peptide portion were less cytotoxic even when they possessed full enzyme inhibitory activity. PMID- 11927258 TI - Yersiniabactin synthetase: a four-protein assembly line producing the nonribosomal peptide/polyketide hybrid siderophore of Yersinia pestis. AB - Yersiniabactin synthetase comprises four proteins, YbtE, HMWP1, HMWP2, and YbtU, encompassing seventeen functional domains, twelve catalytic and five carrier, to select, activate, and incorporate salicylate, three cysteines, and one malonyl moiety into the iron chelator yersiniabactin (Ybt). In the present study, yersiniabactin has been reconstituted in vitro from the 4 protein assembly line by the use of eight biosynthetic precursors. The rate of one turnover, comprising 22 chemical operations performed by the assembly line to release the completed Ybt molecule, was determined at 1.4 min(-1). During the course of Ybt production, the elongating acyl-S-enzyme chain was shown to transfer across a nonribosomal peptide synthetase/polyketide synthase (NRPS/PKS) interprotein interface and then a PKS/NRPS intraprotein interface. This study on the Ybt synthetase assembly line represents the first complete in vitro reconstitution of a nonribosomal peptide/polyketide hybrid system. PMID- 11927257 TI - Heme-artemisinin adducts are crucial mediators of the ability of artemisinin to inhibit heme polymerization. AB - A lack of molecular understanding of the targets and mechanisms of artemisinin action has impeded the improvisation of more efficient antimalarials based on this class of endoperoxide drugs. We have synthesized a heme-artemisinin adduct designated as "hemart" to discover if it mediates the ability of artemisinin to inhibit heme polymerization. Hemart mimics heme in binding to Plasmodium falciparum histidine-rich protein II (PfHRP II) but cannot self-polymerize. Instead, it inhibits all heme polymerizations, including basal and those triggered by PfHRP II, Monooleoyl glycerol (MOG), or P. yoelii extract. Hemart has an edge over heme in displacing heme from PfHRP II, and either low pH or chloroquine dissociates heme but not hemart from PfHRP II. Our results suggest that hemart, by mimicking heme, stalls all mechanisms of heme polymerization, resulting in the death of the malaria parasite. PMID- 11927259 TI - Direct visualization of a DNA glycosylase searching for damage. AB - DNA glycosylases preserve the integrity of genetic information by recognizing damaged bases in the genome and catalyzing their excision. It is unknown how DNA glycosylases locate covalently modified bases hidden in the DNA helix amongst vast numbers of normal bases. Here we employ atomic-force microscopy (AFM) with carbon nanotube probes to image search intermediates of human 8-oxoguanine DNA glycosylase (hOGG1) scanning DNA. We show that hOGG1 interrogates DNA at undamaged sites by inducing drastic kinks. The sharp DNA bending angle of these non-lesion-specific search intermediates closely matches that observed in the specific complex of 8-oxoguanine-containing DNA bound to hOGG1. These findings indicate that hOGG1 actively distorts DNA while searching for damaged bases. PMID- 11927260 TI - GnRH binding RNA and DNA Spiegelmers: a novel approach toward GnRH antagonism. AB - Mirror-image oligonucleotide ligands (Spiegelmers) that bind to the pharmacologically relevant target gonadotropin-releasing hormone I (GnRH) with high affinity and high specificity have been identified using the Spiegelmer technology. GnRH is a decapeptide that plays an important role in mammalian reproduction and sexual maturation and is associated with several benign and malignant diseases. First, aptamers that bind to D-GnRH with dissociation constants of 50-100 nM were isolated out of RNA and DNA libraries. The respective enantiomers of the DNA and RNA aptamers were synthesized, and their binding to L GnRH was shown. These Spiegelmers bind to L-GnRH with similar affinity to that of the corresponding aptamers that bind to D-GnRH. We further demonstrated dose dependent inhibition of GnRH-induced Ca(2+) release in Chinese hamster ovary cells that were stably transfected with the human GnRH receptor. PMID- 11927261 TI - Suppression of DNA-mediated charge transport by BamHI binding. AB - A guanine radical cation produced by one-electron DNA oxidation migrates over long distances through the DNA pi-stack. Fundamental questions regarding the likelihood of charge transport in genomic DNA, the effects of protein binding, and its biological consequences arise as the next issues of study. Electronic effects of protein binding on the efficiency of charge transport were investigated for the endonuclease BamHI-DNA complex. Direct contact of a positively charged guanidium group of BamHI to guanines in the recognition sequence 5'-GGATCC-3' completely suppressed one-electron oxidation of the guanine in the protein binding site and dramatically lowered the charge transport efficiency through the sequence. Electronically insulated guanines, by the hydrogen bonding contact of a guanidium group in BamHI, no longer function as a stepping stone in the charge transport through the DNA pi-stack. PMID- 11927262 TI - Hexaene derivatives of nystatin produced as a result of an induced rearrangement within the nysC polyketide synthase gene in S. noursei ATCC 11455. AB - Genetic manipulation of the polyketide synthase (PKS) gene nysC involved in the biosynthesis of the tetraene antifungal antibiotic nystatin yielded a recombinant strain producing hexaene nystatin derivatives. Analysis of one such compound, S48HX, by LC-MS/MS suggested that it comprises a 36-membered macrolactone ring completely decorated by the post-PKS modification enzymes. Further characterization by bioassay has shown that S48HX exhibits antifungal activity. Genetic analysis of the hexaene-producing mutant revealed an in-frame deletion within the nysC gene via recombination between two homologous ketoreductase domain-encoding sequences. Apparently, this event resulted in the elimination of one complete module from NysC PKS, subsequently leading to the production of the nystatin derivative with a contracted macrolactone ring. These results represent the first example of manipulation of a PKS gene for the biosynthesis of a polyene antibiotic. PMID- 11927263 TI - MgF(3)(-) as a transition state analog of phosphoryl transfer. AB - The formation of complexes between small G proteins and certain of their effectors can be facilitated by aluminum fluorides. Solution studies suggest that magnesium may be able to replace aluminum in such complexes. We have determined the crystal structure of RhoA.GDP bound to RhoGAP in the presence of Mg(2+) and F(-) but without Al(3+). The metallofluoride adopts a trigonal planar arrangement instead of the square planar structure of AlF(4)(-). We have confirmed that these crystals contain magnesium and not aluminum by proton-induced X-ray emission spectroscopy. The structure adopted by GDP.MgF(-) possesses the stereochemistry and approximate charge expected for the transition state. We suggest that MgF3(-) may be the reagent of choice for studying phosphoryl transfer reactions. PMID- 11927264 TI - Two general methods for the isolation of enzyme activities by colony filter screening. AB - We describe two general methodologies, based on filter-sandwich assays, for isolating enzymatic activities from a large repertoire of protein variants expressed in the cytoplasm of E. coli cells. The enzymes are released by the freezing and thawing of bacterial colonies grown on a porous master filter and diffuse to a second "reaction" filter that closely contacts the master filter. Reaction substrates can be immobilized either on the filter or on the enzyme itself (which is then, in turn, captured on the reaction filter). The resulting products are detected with suitable affinity reagents. We used biotin ligase as a model enzyme to assess the performance of the two methodologies. Active enzymes were released by the bacteria, locally biotinylated the immobilized target substrate peptide, and allowed the sensitive and specific detection of individual catalytically active colonies. PMID- 11927265 TI - Using an azobenzene cross-linker to either increase or decrease peptide helix content upon trans-to-cis photoisomerization. AB - Reversible photocontrol of peptide and protein conformation could prove to be a powerful tool for probing function in diverse biological systems. Here, we report reversible photoswitching of the helix content in short peptides containing an azobenzene cross-linker between cysteine residues at positions i, i + 4, or i, i + 11 in the sequence. Trans-to-cis photoisomerization significantly increases the helix content in the i, i + 4 case and significantly decreases the helix content in the i, i + 11 case. These cross-linker designs significantly expand the possibilities for photocontrol of peptide and protein structure. PMID- 11927266 TI - An aPPARently protective mechanism for keratinocytes in wounded skin. PMID- 11927267 TI - APP on the move. PMID- 11927269 TI - A cut above the rest? PMID- 11927268 TI - Searching for clues to premature aging. PMID- 11927271 TI - Gene expression profiling to predict lymphoma outcome. PMID- 11927273 TI - Interleukin-5 and eosinophils as therapeutic targets for asthma. AB - Extensive clinical investigations have implicated eosinophils in the pathogenesis of asthma. In a recent clinical trial, humanized monoclonal antibody to interleukin (IL)-5 significantly limited eosinophil migration to the lung. However, treatment did not affect the development of the late-phase response or airways hyperresponsiveness in experimental asthma. Although IL-5 is a key regulator of eosinophilia and attenuation of its actions without signs of clinical improvement raises questions about the contribution of these cells to disease, further studies are warranted to define the effects of anti-IL-5 in the processes that lead to chronic asthma. Furthermore, eosinophil accumulation into allergic tissues should not be viewed as a process that is exclusively regulated by IL-5 but one in which IL-5 greatly contributes. Indeed, data on anti-IL-5 treatments (human and animal models) are confounded by the failure of this approach to completely resolve tissue eosinophilia and the belief that IL-5 alone is the critical molecular switch for eosinophil development and migration. The contribution of these IL-5-independent pathways should be considered when assessing the role of eosinophils in disease processes. PMID- 11927274 TI - Molecular basis of cholesterol homeostasis: lessons from Tangier disease and ABCA1. AB - High-density lipoproteins (HDLs) play a role in transporting cholesterol from peripheral tissues to the liver for elimination from the body. Two hallmarks of cardiovascular disease are the presence of sterol-laden macrophages in the artery wall and reduced plasma HDL levels. A cell-membrane protein called ABCA1 mediates the secretion of excess cholesterol from cells into the HDL metabolic pathway. Mutations in ABCA1 cause Tangier disease, a severe HDL deficiency syndrome characterized by accumulation of cholesterol in tissue macrophages and prevalent atherosclerosis. Because of its ability to deplete macrophages of cholesterol and to raise plasma HDL levels, ABCA1 has become a promising therapeutic target for preventing cardiovascular disease. PMID- 11927275 TI - The coming of age of 1,25-dihydroxyvitamin D(3) analogs as immunomodulatory agents. AB - The active form of vitamin D, 1,25-dihydroxyvitamin D(3)[1,25(OH)(2)D(3)], is a secosteroid hormone that regulates calcium and bone metabolism, controls cell proliferation and differentiation, and exerts immunoregulatory activities. This range of functions has been exploited clinically to treat a variety of conditions, from secondary hyperparathyroidism to osteoporosis, to autoimmune diseases such as psoriasis. Recent advances in understanding 1,25(OH)(2)D(3) functions and novel insights into the mechanisms of its immunomodulatory properties suggest a wider applicability of this hormone in the treatment of autoimmune diseases and allograft rejection. PMID- 11927277 TI - Carbohydrate-binding proteins in cancer, and their ligands as therapeutic agents. AB - Experimental evidence directly implicates complex carbohydrates in recognition processes, including adhesion between cells, adhesion of cells to the extracellular matrix, and specific recognition of cells by one another. In addition, carbohydrates are recognized as differentiation markers and as antigenic determinants. Lectins are nonenzymatic proteins present in plants and animals, which preferentially bind to specific carbohydrate structures and play an important role in cell recognition. Modified carbohydrates and oligosaccharides have the ability to interfere with carbohydrate-protein interactions and therefore, inhibit the cell-cell recognition and adhesion processes, which play an important role in cancer growth and progression. Carbohydrate ligands therefore, are candidates to play important roles in cancer therapeutics. PMID- 11927276 TI - Caretaker tumour suppressor genes that defend genome integrity. AB - Cancers arise as a result of genetic changes that impact upon cell proliferation through promoting cell division and/or inhibiting cell death. Tumour suppressor (TS) genes are the targets for many of these genetic changes. In general, both alleles of TS genes must be disrupted to observe a phenotypic effect. Broadly speaking, there are two types of TS gene: 'gatekeepers' and 'caretakers'. In contrast to gatekeepers, caretaker genes do not directly regulate proliferation, but act to prevent genomic instability. Thus, mutation of caretaker genes leads to accelerated conversion of a normal cell to a neoplastic cell. Many caretaker genes are required for the maintenance of genome integrity. This review focuses on those caretaker genes that play a role, directly or indirectly, in the repair of DNA strand breaks by the homologous recombination pathway, and that are associated with cancer-prone clinical syndromes, in particular ataxia telangiectasia, hereditary breast cancer, Bloom's syndrome and Werner's syndrome. PMID- 11927279 TI - Translating basic cancer research into new cancer therapeutics. PMID- 11927280 TI - Histone modification: the 'next wave' in cancer therapeutics. PMID- 11927281 TI - Aromatase inhibitors continue their ATAC on tamoxifen. PMID- 11927282 TI - STI571 (Gleevec) as a paradigm for cancer therapy. AB - STI571 (Gleevec, imatinib mesylate) exemplifies the successful development of a rationally designed, molecularly targeted therapy for the treatment of a specific cancer. This article reviews the identification of Bcr-Abl as a therapeutic target in chronic myelogenous leukemia and the steps in the development of an agent to inactivate this abnormality. Issues related to clinical trials of molecularly targeted agents are discussed, including dose and patient selection and possible mechanisms of resistance to STI571. Finally, the potential use of STI571 with different tumors and the translation of this paradigm to other malignancies are explored. PMID- 11927283 TI - The ErbB receptor family: a therapeutic target for cancer. AB - This article reviews the current state of efforts targeting the ErbB family of tyrosine kinase receptors in cancer therapy. In particular, preliminary results will be discussed of studies of the first generation of therapeutics to enter clinical evaluation in malignant diseases. Results of recently conducted clinical studies with ZD1839 (Iressa), OSI-774 (Tarceva), Cetuximab (IMC-C225) and trastuzumab (Herceptin) and several other compounds are presented. Potential advantages and disadvantages of these different therapeutic modalities, as well as future challenges of evaluating ErbB-targeted agents in the clinic, are presented. PMID- 11927284 TI - Unraveling the complexities of the Raf/MAP kinase pathway for pharmacological intervention. AB - The Ras-MAP kinase pathway has attracted much attention from academic and pharmaceutical laboratories because of its central role in regulating tumor cell growth and survival, differentiation and angiogenesis. Although the central players in this pathway -Ras, Raf, and MEK - have been well studied, how best to exploit them for therapeutic gain has eluded oncology researchers in the past. Several small-molecule inhibitors that target specific steps of the MAP kinase cascade have recently entered the clinical arena. While we await answers on their ultimate therapeutic use, the availability of translational assays for monitoring target suppression will no doubt play a significant role in optimizing our chances of success. PMID- 11927285 TI - Complexities in the development of cyclin-dependent kinase inhibitor drugs. AB - Abnormalities in the normal regulation of the cell cycle are a hallmark of neoplasia. Drugs directed against the cyclin-dependent kinases (CDKs), which govern the normal orderly progression through the cell cycle, have been proposed to address the pathogenic defect in tumors. Recently, CDK family members that do not regulate the cell cycle directly but instead influence transcription (CDK7, CDK8, and CDK9) and neuronal and secretory cell function (CDK5) have been described. Continued synthetic chemistry efforts have defined important new selective inhibitors of CDKs, and strategies directed at newly described CDK related targets, such as transcription control, can now be envisaged. CDKs remain important and novel targets whose potential needs to be more fully explored, albeit in light of the newly emerging complexities of their cellular physiology. PMID- 11927286 TI - Therapeutic exploitation of the p53 pathway. AB - Analysis of the gene encoding p53 could serve to evaluate the effectiveness of a cancer treatment. Mutations in this gene occur in half of all human cancers, and regulation of the protein is defective in a variety of others. Novel strategies that exploit our knowledge of the function and regulation of p53 are being actively investigated. Strategies directed at treating tumours that have p53 mutations include gene therapy, viruses that only replicate in p53 deficient cells, and the search for small molecules that reactivate mutant p53. Potentiating the function of p53 in a non-genotoxic way in tumours that express wildtype protein can be achieved by inhibiting the expression and function of Mdm2 or viral oncoproteins. PMID- 11927287 TI - Epigenomics and epigenetic therapy of cancer. AB - Epigenetic inactivation of genes that are crucial for the control of normal cell growth is a hallmark of cancer cells. These epigenetic mechanisms include crosstalk between DNA methylation, histone modification and other components of chromatin higher-order structure, and lead to the regulation of gene transcription. Re-expression of genes epigenetically inactivated can result in the suppression of tumour growth or sensitization to other anticancer therapies. Small molecules that reverse epigenetic inactivation are now undergoing clinical trials in cancer patients. This, together with epigenomic analysis of chromatin alterations such as DNA methylation and histone acetylation, opens up the potential both to define epigenetic patterns of gene inactivation in tumours and to use drugs that target epigenetic silencing. PMID- 11927288 TI - Proteasome inhibition: a novel approach to cancer therapy. AB - The central role of the proteasome in controlling the expression of regulators of cell proliferation and survival has led to interest in developing proteasome inhibitors as novel anticancer agents. In vitro and in vivo studies have shown that proteasome inhibitors have activity against a variety of tumor types. One of these agents, PS-341, has been tested in phase I trials in a variety of tumor types; in these trials, PS-341 treatment was well tolerated and preliminary evidence of biological activity was observed in some patients. Phase II trials in several hematological malignancies and solid tumor types are now in progress. PMID- 11927289 TI - Hsp90 inhibitors as novel cancer chemotherapeutic agents. AB - Heat shock protein 90 (Hsp90) is a molecular chaperone whose association is required for the stability and function of multiple mutated, chimeric and over expressed signaling proteins that promote the growth and/or survival of cancer cells. Hsp90 client proteins include mutated p53, Bcr-Abl, Raf-1, Akt, ErbB2 and hypoxia-inducible factor 1 alpha (HIF-1 alpha). Hsp90 inhibitors, by interacting specifically with a single molecular target, cause the destabilization and eventual degradation of Hsp90 client proteins, and they have shown promising antitumor activity in preclinical model systems. One Hsp90 inhibitor, 17 allylaminogeldanamycin (17AAG), is currently in phase I clinical trial. Because of the chemoprotective activity of several proteins that are Hsp90 clients, the combination of an Hsp90 inhibitor with a standard chemotherapeutic agent could dramatically increase the in vivo efficacy of the therapeutic agent. PMID- 11927290 TI - HIF-1 and tumor progression: pathophysiology and therapeutics. AB - Hypoxia-inducible factor 1 (HIF-1) controls oxygen delivery (via angiogenesis) and metabolic adaptation to hypoxia (via glycolysis). HIF-1 consists of a constitutively expressed HIF-1 beta subunit and an oxygen- and growth-factor regulated HIF-1 alpha subunit. In xenografts, tumor growth and angiogenesis are correlated with HIF-1 expression. In human cancers, HIF-1 alpha is overexpressed as a result of intratumoral hypoxia and genetic alterations affecting key oncogenes and tumor suppressor genes. HIF-1 alpha overexpression in biopsies of brain, breast, cervical, esophageal, oropharyngeal and ovarian cancers is correlated with treatment failure and mortality. Increased HIF-1 activity promotes tumor progression, and inhibition of HIF-1 could represent a novel approach to cancer therapy. PMID- 11927291 TI - The emerging fields of suicide gene therapy and virotherapy. AB - Gene therapy is defined as a technology aimed at modifying the genetic component of cells for therapeutic benefit. 'Suicide genes' can be introduced into cancer cells to make them more sensitive to chemotherapeutics or toxins. Chemotherapeutic suicide gene therapy approaches are known as gene-directed enzyme prodrug therapy or gene-prodrug activation therapy. Other approaches include replacement gene therapy, antisense strategies and induction of resistance to normal cells. All gene therapy strategies share a common component, which is the need for a selective delivery vehicle or vector with tumor-targeting capabilities. This need has led to the in-depth investigation of viruses as new vectors for gene therapy. PMID- 11927292 TI - Postoperative ileus after abdominal surgery. PMID- 11927293 TI - Detrusor apron, associated vascular plexus, and avascular plane: relevance to radical retropubic prostatectomy--anatomic and surgical commentary. PMID- 11927295 TI - Efficacy of desmopressin in treatment of refractory nocturia in patients older than 65 years. AB - OBJECTIVES: To evaluate the efficacy of desmopressin treatment in patients 65 years old and older with nocturia and to determine whether baseline urodynamic characteristics influenced the outcome of treatment. METHODS: Patients with nocturia three or more times a night and nocturnal polyuria refractory to medication were treated with oral desmopressin 0.1 mg at bedtime for 4 weeks. Data from urodynamic studies and a voiding diary, nocturnal urine volume, urine specific gravity, serum sodium and potassium level, and quality of life index were measured at baseline, 4 weeks, and 4 weeks after discontinuation of treatment. RESULTS: A total of 30 patients (25 men and 5 women) were enrolled in the study. The mean age was 75.4 +/- 6.6 years. Five patients (16.7%) reported side effects, including hyponatremia in one. Twenty patients (66.7%) reported a good response with both reduced nocturnal frequency (5.2 +/- 1.16 times versus 2.24 +/- 1.12 times a night, P = 0.000) and urine volume (955.6 +/- 255.9 mL versus 522.8 +/- 210.5 mL, P <0.0001). Two patients (6.7%) had improved nocturnal frequency, and 3 patients (10%) reported no effect at all. After discontinuing the medication for 4 weeks, 13 patients (52%) had improved symptoms compared with baseline and 6 (24%) remained at their post-treatment frequency of nocturia. Urodynamic studies revealed that 15 patients had detrusor instability and 17 had a cystometric capacity of 250 mL or less. No significant difference was found in the success rate relative to the urodynamic results. CONCLUSIONS: Desmopressin is safe and effective in the treatment of severe nocturia in patients 65 years old and older. PMID- 11927294 TI - Neoadjuvant hormonal therapy and older age are associated with adverse sexual health-related quality-of-life outcome after prostate brachytherapy. AB - OBJECTIVES: Brachytherapy is increasingly used as a treatment for localized prostate cancer but information regarding long-term, postimplantation, patient reported sexual health-related quality-of-life (HRQOL) is scant. Neoadjuvant hormonal therapy is commonly administered with brachytherapy, yet its potentially adverse effects on subsequent sexual health have not been described using a validated HRQOL instrument. We used a validated HRQOL survey to characterize the significance of neoadjuvant hormonal therapy and other baseline factors on postimplantation sexual function and impairment. METHODS: A cross-sectional survey using the expanded prostate cancer index composite HRQOL instrument was administered to all 114 localized prostate cancer patients who underwent ultrasound-guided, transperineal brachytherapy during a 4-year period and to 142 age-matched control men. Multivariable models measured the association of baseline factors and covariates with postimplantation sexual HRQOL. RESULTS: Older age (P = 0.01) and neoadjuvant hormonal therapy (P = 0.009) were independently associated with diminished sexual HRQOL after prostate brachytherapy. Among patients younger than 69 years old, 33% reported at least fair sexual function after brachytherapy alone compared with 19% of men after brachytherapy with neoadjuvant hormonal therapy. Of the age-matched control men younger than 69 years old, 78% reported at least fair sexual function. Among patients older than 69 years, 26% reported at least fair sexual function after brachytherapy alone compared with 5% after brachytherapy with neoadjuvant hormonal therapy, and 61% of age-matched controls reported at least fair sexual function. CONCLUSIONS: Patient age and neoadjuvant hormonal therapy are independent, significant determinants of sexual HRQOL after prostate brachytherapy. These factors should be taken into consideration when counseling patients with localized prostate cancer regarding the expected, postimplantation sexual HRQOL outcome. PMID- 11927296 TI - Changing indications of open stone surgery. AB - OBJECTIVES: To compare the current role of open stone surgery at our institution to previously reported data. In 1989, the indications for open surgical treatment of urinary calculi at our institution were reviewed. In the intervening years, tremendous advances have been made in minimally invasive treatment of urinary calculi. METHODS: A retrospective evaluation of all patients undergoing procedures for the purpose of stone removal or fragmentation at Wake Forest University Baptist Medical Center between January 1, 1998 and May 31, 2001 was conducted. This was compared with data reported from our institution describing similar procedures in the first 19 months after introduction of the Dornier HM3 lithotriptor. RESULTS: Of 986 procedures performed for the purpose of stone removal or fragmentation between January 1, 1998 and May 31, 2001, 0.7% were open surgical procedures. Of these procedures, 85.8% were performed for anatomic indications. Patients referred from other institutions for evaluation after endoscopic treatment failure comprised the remaining 14.2% of this group. In the previously reported data, 893 procedures were performed for the treatment of urinary calculi, of which 4.1% were open operations. Of these patients, 48.6% underwent an open surgical procedure after unsuccessful endoscopic treatment of urinary calculi, and 48.7% of these patients underwent open surgery for anatomic indications. CONCLUSIONS: Open surgical stone removal remains a viable treatment option for select patients. Technologic advances and improved surgical skills have greatly reduced the number of patients requiring open surgery. This approach is mainly used for patients with complex calculous disease associated with anatomic abnormalities. PMID- 11927299 TI - Laparoscopic pyeloplasty for UPJ obstruction with crossing vessels: contrast enhanced color Doppler findings and long-term outcome. AB - OBJECTIVES: To evaluate, in the present long-term follow-up study, contrast enhanced color Doppler imaging (CDI) findings and the clinical outcome of patients with crossing vessels at the obstructed ureteropelvic junction (UPJ), who underwent laparoscopic pyeloplasty. In a previous study, contrast-enhanced CDI proved capable of detecting crossing vessels at the UPJ. METHODS: A total of 23 patients, who had undergone laparoscopic pyeloplasty and displacement of crossing vessels for UPJ obstruction at least 2 years before this study (mean 27 months), underwent contrast-enhanced CDI, intravenous urography, and renography. Contrast-enhanced CDI was performed using intravenously administered Levovist to assess the displacement of the vessels relative to the UPJ. All patients completed analog follow-up pain scales and quality-of-life assessment questionnaires. RESULTS. Contrast-enhanced CDI revealed a cranial displacement (mean 1.3 cm) of the crossing vessels from the UPJ in all 23 cases. Intravenous urography showed a decrease in the degree of hydronephrosis, with a success rate of 100% in low-grade and 86% in high-grade hydronephrosis. The split renal function improved from 39.7% to 48.1%. Analog pain scale measurements demonstrated a mean improvement in pain of 92% (range 73% to 100%) and a mean quality-of-life score of 94 (range 78 to 100). CONCLUSIONS: Our series of patients with crossing vessels at the UPJ treated by laparoscopic pyeloplasty showed an excellent long-term successful outcome. Contrast-enhanced CDI allows for preoperative detection, as well as postoperative assessment, of the displacement of the crossing vessel. We recommend that the presence of a crossing vessel be routinely determined preoperatively, because it may influence the choice of treatment modality and thereby the clinical outcome. PMID- 11927298 TI - Clinical association with urinary glycosaminoglycans and urolithiasis. AB - OBJECTIVES: To determine the clinical association between urinary glycosaminoglycan (GAG) concentration and kidney stone disease. METHODS: Thirty five patients (14 women and 21 men) with a history of stone disease and 37 controls (13 women and 24 men) were evaluated for urinary GAG concentration. By using a new dye-binding assay, the total GAG concentration in the urine was measured and corrected to urinary creatinine levels (micrograms of GAG per milligram creatinine). RESULTS: The mean urinary GAG concentration in those with stones was significantly lower (31.5 +/- 2.6 microg GAG/mg creatinine) than in the controls (43.8 +/- 3.8 microg GAG/mg creatinine, P = 0.01). Male patients with stones also had a significantly lower mean GAG concentration (26.1 +/- 1.8) than did the female patients (39.6 +/- 5.3, P = 0.009). The mean GAG concentration between ureteral (n = 13) versus renal (n = 22), single (n = 19) versus multiple (n = 16), family history (n = 11) versus no family history (n = 24), large (n = 13) versus small (n = 20), and the presence (n = 22) versus absence (n = 13) of residual stones did not show any significant differences. However, patients with recurrent stone formation (n = 21) had significantly lower mean GAG levels (26.4 +/- 1.6) compared with those with single stone formation (n = 14; 39.2 +/- 5.5, P = 0.01). CONCLUSIONS: Lower urinary GAG levels are more common in patients with stone formation. This may play a more determinant role in male patients and those with recurrent stone formation. PMID- 11927300 TI - Recoverability of renal function after relief of acute complete ureteral obstruction: clinical prospective study of the role of renal resistive index. AB - OBJECTIVES: To study the values of the renal resistive index (RI) before and at different points after relief of obstructive anuria and to correlate these values with the corresponding values of serum creatinine and with the recovery of renal function after release of obstruction. METHODS: A total of 32 consecutive patients with obstructive anuria were prospectively evaluated by measurement of RI before drainage and at 3 days, 1 week, 2 weeks, and 4 weeks after drainage. Serum creatinine was measured at all points of the RI examination. Moreover, RI was measured in an age and sex-matched control group of 24 consecutive healthy donors and volunteers. RESULTS: The study included 40 obstructed and 48 normal kidneys. In the obstructed kidneys, the mean RI values decreased significantly from 0.78 +/- 0.05 before drainage to 0.70 +/- 0.09 at 3 days after drainage (P <0.001) with an additional significant reduction to 0.68 +/- 0.08 at 7 days after drainage (P <0.01) and stabilized thereafter. Serum creatinine decreased significantly from 8.4 +/- 4.4 mg/dL before drainage to 4.7 +/- 3.8 mg/dL 3 days after drainage (P <0.001) and then to 3.6 +/- 3.7 mg/dL 7 days after drainage (P <0.001) and stabilized thereafter. The correlation between the RI and serum creatinine at the overall points of measurement was good. Obstructed kidneys were classified into two groups according to the recovery of renal function after obstruction relief: those that showed significant improvement of serum creatinine (24 kidneys, group 1) and those with no significant improvement of serum creatinine (16 kidneys, group 2). In group 1, the difference between the mean RI values before and after drainage was statistically significant (0.78 +/- 0.05 versus 0.64 +/- 0.06, P <0.001); in group 2, the difference between the before and after drainage RI values was not significant (0.781 +/- 0.040 versus 0.779 +/ 0.039). The mean RI of the normal kidneys was 0.66 +/- 0.04. A comparison between the mean RI values of the control group and the mean RI values of the obstructed patients after drainage showed no significant difference in group 1; markedly higher values were noted in group 2 at all points after drainage. CONCLUSIONS: In the setting of acute complete renal obstruction, the RI has a good positive correlation with serum creatinine. Recovery of renal function could not be predicted from the changes in RI before obstruction release. However, a reversal of a previously elevated RI could be used as an early indicator that renal function recovery is likely. PMID- 11927301 TI - Characterization of urinary symptoms in patients with ureteral stents. AB - OBJECTIVES: To prospectively assess the prevalence and bother of various urinary tract symptoms caused by indwelling ureteral stents using validated questionnaires. METHODS: The study consisted of 60 patients with unilateral ureteral stents. Of these, 30 patients were asked to complete the International Prostate Symptoms Score questionnaire, with additional questions on dysuria, hematuria, and loin pain. The remaining 30 patients were asked to complete the International Continence Society questionnaire. These patients were also asked to complete frequency volume charts and undergo uroflowmetry studies. The questionnaires were completed with a stent in situ and 6 weeks after its removal. RESULTS: Forty-eight patients (36 men and 12 women, mean age 52.8 years) completed the study. A large proportion (80%) of patients reported one or more urinary symptoms. Analysis of the IPSS data revealed impaired global quality of life owing to these urinary symptoms. The responses to additional questions on dysuria and hematuria showed a statistically significant difference, but most International Prostate Symptoms Score questions did not. The results of the International Continence Society study identified storage symptoms, incontinence (60%), and bladder pain (80%) as important bothersome problems. The results of the frequency volume charts were in agreement with the storage symptoms. CONCLUSIONS: Patients with indwelling ureteral stents have a wide range of urinary symptoms that affect their quality of life. Storage symptoms, bladder pain, and hematuria pose a major bother. None of the existing questionnaires covered the entire range of symptoms. The results are useful in better understanding the urinary symptoms associated with stents and in providing patient counseling. PMID- 11927302 TI - Prevalence and epidemiologic characteristics of urolithiasis in Seoul, Korea. AB - OBJECTIVES: To study the epidemiologic details of urolithiasis in Seoul, Korea. The prevalence of urolithiasis varies by different regions in the world and has been reported to be 4% to 15%. METHODS: A total of 2643 persons 40 to 79 years old living in Seoul were selected by stepwise random sampling. Questionnaires were used and completed by trained interviewers concerning whether the subjects had experienced urinary stone disease at any time in their life. RESULTS: Of 2643 persons interviewed, 65 experienced 78 episodes of urinary stone formation in their lifetime. Calibrated after the Seoul population census, the overall standardized lifetime prevalence rate was estimated to be 3.5%, 6.0% in men and 1.8% in women. The point prevalence rate was estimated to be 0.9%, 1.9% in men and 0.3% in women. Most of the urinary stone diseases were diagnosed at the fifth decade. Kidney stones in family members were present in 8.1% of patients with stones and 4.1% of stone-free control individuals. Twenty percent of the patients with stones experienced recurrences. CONCLUSIONS: In this first systematically assessed epidemiologic survey about the rate of urolithiasis in Korea, 6.0% of Korean men and 1.8% of women are expected to experience urinary stone disease during their lifetime. PMID- 11927303 TI - Effect of percutaneous nephrolithotomy on differential renal function as measured by mercaptoacetyl triglycine nuclear renography. AB - OBJECTIVES: To determine the impact on differential renal function of percutaneous nephrolithotomy for complex renal calculi. METHODS: From July 1999 to December 2000, 45 patients underwent percutaneous nephrolithotomy. Of these, 19 agreed to participate in the study. All patients completed a quantitative assessment of differential renal function preoperatively and postoperatively with technetium 99m mercaptoacetyl triglycine nuclear renography and serum creatinine measurements. RESULTS: The mean patient age was 49 years (range 11 to 75) for the 13 female and 6 male patients. The mean stone burden was 1432 mm(2) (range 156 to 5220). The mean surgical time was 2.57 hours (range 1.17 to 5.08). The median hospital stay was 2.0 days (range 1 to 19). Of the 19 patients, 13 (68%) were stone free after one procedure. Four patients underwent ureteroscopy with stone extraction for residual fragments. One patient underwent secondary extracorporeal shock wave lithotripsy. One patient underwent nephrectomy for poor renal function. Renal function, for the entire group, increased from 36.8% preoperatively to 38.5% postoperatively. Renal function was preserved in 16 (84%) of 19 patients, including improvement of function in 7 (37%) of 19 patients. Serum creatinine was unchanged in the two groups. CONCLUSIONS: Percutaneous nephrolithotomy does not result in loss of renal function when treating complex renal calculi as measured by nuclear scintigraphy. Operative and hospitalization times were decreased compared with historical open nephrolithotomy and stone clearance was similar. PMID- 11927306 TI - Tubeless percutaneous renal surgery: review of first 112 patients. AB - OBJECTIVES: To assess the outcome and safety of the first 112 patients undergoing tubeless percutaneous renal surgery. METHODS: A total of 116 renal units in 112 patients underwent tubeless percutaneous renal surgery from December 1995 to November 2000 performed by a single urologist. The "tubeless" procedures consisted of nephrolithotripsy or endopyelotomy. An antegrade internal ureteral stent was placed during surgery, as was a Foley catheter for bladder drainage, in all patients. Nephrostomy tubes were not used in any patient. The incidence of complications, length of hospitalization, outcomes, transfusion requirements, stone burden, and stone-free rates were obtained by chart review. RESULTS: Of the 112 patients, 86 patients underwent percutaneous stone extraction and 26 patients underwent percutaneous antegrade endopyelotomy. In the group of patients undergoing percutaneous stone extraction, the mean stone burden was 3.30 cm(2). Patients stayed in the hospital an average of 1.25 days. A total of 5 patients required postoperative transfusions. One patient was found on readmission to have a pseudoaneurysm. A 93.0% stone-free rate was achieved. The correlations between transfusion rates, days of hospitalization, and outcome with stone burden were not statistically significant (P = 0.56, 0.78, and 0.18, respectively). For the endopyelotomy group, the mean length of hospitalization was 1.56 days. Only 1 patient required a transfusion, and 3 patients had postoperative complications. Of the 26 patients, 88.5% had a successful outcome from their endopyelotomy. CONCLUSIONS: Tubeless percutaneous renal surgery is a safe and effective procedure and can be performed in patients even with a moderate stone burden. PMID- 11927308 TI - Prognostic features of pathologic stage T1 renal cell carcinoma after radical nephrectomy. AB - OBJECTIVES: To assess the effect of renal cell carcinoma (RCC) subtype, tumor size, and Fuhrman grade on clinical outcome in patients with pathologic T1 (pT1) RCC treated with radical nephrectomy. METHODS: Between 1970 and 1998, 840 patients underwent radical nephrectomy for pT1 RCC. Tumors were subtyped and graded. Univariate and multivariate Cox proportional hazards models were fitted to assess the features associated with metastasis-free survival (MFS) and cancer specific survival (CSS). We identified a range of tumor sizes of clear cell RCC in which a transition occurred from low to high risk. Cox proportional hazards models were then fitted by using size cutoffs. RESULTS: The mean follow-up (+/- SD) was 9.4 +/- 6.6 years among the patients alive at latest follow-up. At 10 years, the CSS and MFS for clear cell RCC (n = 682) were 89.1% and 88.6%, respectively; for papillary RCC (n = 122), they were 95.5% and 93.8%; and for chromophobe RCC (n = 33), they were both 100%. The differences in CSS (P = 0.013) and MFS (P = 0.023) between clear cell RCC and the other subtypes were statistically significant. For clear cell RCC, tumor size and Fuhrman grade were independently associated with CSS and MFS (P <0.001). A transition in risk occurred for tumor sizes between 4.5 and 5.0 cm, and the tumor size cutoff of 5.0 cm had the highest concordance index for predicting CSS and MFS. CONCLUSIONS: RCC subtype is a strong independent prognostic variable for patients with pT1 RCC treated with radical nephrectomy. For clear cell RCC, Fuhrman grade and tumor size are independently associated with CSS and MFS. PMID- 11927310 TI - Artificial urinary sphincter placement in men after cystectomy with orthotopic ileal neobladder: continence, complications, and quality of life. AB - OBJECTIVES: To review our experience regarding the overall efficacy and safety of an artificial urinary sphincter (AUS) in men with stress urinary incontinence (SUI) after cystoprostatectomy with an orthotopic ileal neobladder. METHODS: We performed a retrospective review of 5 men who underwent placement of an AUS (AMS 800) for severe SUI after radical cystectomy with an orthotopic ileal neobladder. Incontinence symptoms and quality of life were quantified using two validated continence questionnaires (Urogenital Distress Inventory Short Form and Incontinence Impact Questionnaire Short Form) and a brief addendum questionnaire. The degree of continence, perioperative and postoperative complications and infections, symptom distress, quality of life, and patient satisfaction were compared before and after AUS placement. RESULTS: Complete (0 pads per day) or social (1 pad or less per day) continence was reported in 5 of 5 patients after AUS placement. The average pad usage significantly decreased from 6.2 to 0.6 per day (P <0.001). No significant perioperative or postoperative complications or infections were noted. Symptom distress, quality of life, and patient satisfaction were significantly improved after AUS placement (P < 0.01, < 0.0001, and < 0.001, respectively). CONCLUSIONS: With short-term follow-up and a limited number of patients, the placement of an AUS for treatment of SUI after bladder substitution is well tolerated and reliable and has a positive impact on patients' quality of life. PMID- 11927309 TI - Technique of combined pubovaginal sling and cystocele repair using a single piece of cadaveric dermal graft. AB - OBJECTIVES: To investigate the feasibility of using a single piece of cadaveric dermal allograft for the repair of stress urinary incontinence (SUI) with concurrent cystocele. METHODS: Nineteen patients with combined SUI and symptomatic grade III cystoceles were treated. Eleven of 19 patients had undergone prior repairs for SUI. All patients underwent a combined pubovaginal sling procedure and cystocele repair using a single piece of cadaveric dermal allograft (3 x 7 cm). The single strip of dermal graft was placed in a longitudinal direction along the anterior vagina. The distal segment of the allograft supported the urethra, and the proximal portion supported the central cystocele defect and was sutured to the pubocervical fascia. The mean follow-up was 28 +/- 4 months and patients were monitored by physical examination, videourodynamic studies, and completion of the bladder bothersome visual analog scale. RESULTS: Of the 19 patients, 1 developed an acute infection and failure of the graft after presenting with fever, discharge, dysuria, and incontinence. The autolysed graft was removed, and she subsequently underwent successful autologous fascial repair. Of the remaining 18 patients, 17 were cured of their SUI, including 10 who had had prior repairs, and 16 had no recurrence of cystocele and 2 had asymptomatic grade I and II cystoceles. One patient developed de novo detrusor instability that was successfully treated with anticholinergic medication. No cases of urethral obstruction occurred. CONCLUSIONS: Although the follow-up was short, the use of a single piece of cadaveric dermal graft slings for concomitant pubovaginal sling and cystocele repair is feasible and simple to perform. At more than 2 years of follow-up, documented by videourodynamic studies, neither urethral obstruction nor symptomatic cystocele recurrence was found. PMID- 11927311 TI - Accuracy of family history of cancer as reported by men with prostate cancer. AB - OBJECTIVES: To report on the accuracy of probands providing information on specific cancer types in their families and the ability of investigators to document these reports. Accurate information on the health status of family members is critical when studying familial patterns of diseases. However, collecting these data require significant resources. METHODS: We identified 143 patients with prostate cancer from the University of Texas M. D. Anderson Cancer Center who had reported at least 1 first-degree relative with cancer. There were 263 first-degree relatives identified, for whom we confirmed diagnoses using medical records, death certificates, and verbal confirmation. The data are reported in summary statistics and compared with chi-square analysis. RESULTS: We documented 78% of the reports, with an accuracy rate of 81.6%. We found that accuracy was highly related to the site reported. Accuracy and documentation levels were not related to the age or income of the proband. The education level was significantly associated with the ability to document cancer, but not with the accuracy of the report. The accuracy and documentation differed by the relationship of the first-degree relative to the proband. CONCLUSIONS: Proband reporting of cancer in first-degree relatives varies widely by site, with common metastatic sites the most inaccurate. No reliable demographic factors were found that would reasonably predict the ability to document the accuracy of the report. We found a significant proportion of proband-reported prostate cancer was, in reality, benign prostatic hyperplasia. We propose a strategy of targeting male relatives and reports of cancer in common metastatic sites for aggressive follow up. PMID- 11927312 TI - Clinical significance of biopsy-derived primary Gleason score among radical prostatectomy candidates with Gleason 7 tumors. AB - OBJECTIVES: To determine the clinical value of assessing primary Gleason scoring among patients with grade 7 tumors at needle biopsy with emphasis on its correlation with final pathologic stage and grade. METHODS: A total of 108 contemporary cases of surgically treated prostate cancer graded as Gleason score 7 by needle biopsy were reviewed. The influence of primary Gleason pattern, as determined by needle biopsy, on pathologic outcomes, including positive surgical margins and extracapsular extension, was assessed. We also examined the positive predictive value of the assigned primary Gleason pattern. RESULTS: Among the 108 cases, 79 (73.1%) and 29 (26.9%) demonstrated primary pattern 3 and 4, respectively. The positive predictive value of primary Gleason 3 and 4 was 46% and 35%, respectively. When stratified by the needle biopsy primary Gleason pattern, the incidence of extracapsular extension, positive surgical margins, and seminal vesicle involvement was not significantly different. CONCLUSIONS: Pathologic features at radical prostatectomy among patients with biopsy-derived Gleason 7 tumors are not associated with the assigned primary pattern. These data call into question whether pathologists should issue a primary score among patients with Gleason 7 cancers at needle biopsy. PMID- 11927313 TI - Implications of radioactive seed migration to the lungs after prostate brachytherapy. AB - OBJECTIVES: To review the incidence and the impact of pulmonary seed migration after prostate brachytherapy on lung function. Isolated reports of seed migration to the lungs after prostate brachytherapy have been published; however, the clinical consequences of this pulmonary migration have not been adequately evaluated. METHODS: We performed a retrospective review of patients undergoing prostate brachytherapy from 1998 to 2000. Pulmonary imaging with chest x-ray was performed 15 to 90 days (median 45) after the procedure. The chest radiographs were reviewed by a single radiologist (V.S.D.), and patients with seed migration to the lungs were evaluated by a single pulmonologist (A.M.H.) using a questionnaire, chest radiography, and pulmonary function tests. Computed tomography of the prostate after seed implantation was performed to check seed position and dosimetry. Odds ratio, confidence intervals, chi-square tests, and logistic regression analysis were performed to evaluate the relationship between the type and number of seeds used, the incidence of pulmonary migration, and the effects on pulmonary function. RESULTS: A total of 83 patients underwent prostate brachytherapy during the study period and 58 patients underwent chest radiography. Seed migration occurred in 21 (36.2%) of 58 patients (95% confidence interval 23.8% to 48.6%). Thirty-four (0.71%) of 4755 seeds used migrated to the lungs. Nine patients had single seed migration to the right lung and three to the left lung. Nine patients had multiple (maximum of 4) and bilateral seed migration. No consistent relationship was found between seed migration and the type of seeds used (P = 0.24). Borderline statistical significance suggested an increased incidence of seed migration with an increasing number of implanted seeds (P = 0.054). Repeat chest radiography in 21 patients revealed no delayed migration at a median follow-up of 16 months. Clinical and pulmonary function testing revealed no consistent abnormality attributable to seed migration. CONCLUSIONS: Radioactive seed migration occurred in 36.2% of brachytherapy patients who had chest radiographs done in our series. Pulmonary seed migration may be influenced by the number of implanted seeds and does not appear to be influenced by the seed type. Additional study of this observed phenomenon is required. A thorough pulmonary workup failed to reveal any short-term harmful side effects; however, long-term follow-up is needed. Healthcare providers should discuss the possibility of pulmonary seed migration with patients with prostate cancer considering prostate brachytherapy. PMID- 11927314 TI - Predicting disease recurrence in intermediate and high-risk patients undergoing radical prostatectomy using percent positive biopsies: results from CaPSURE. AB - OBJECTIVES: To determine whether percent positive biopsies could be used to predict the probability of disease recurrence in contemporary patients undergoing radical prostatectomy in community-based practice settings. Previous studies have demonstrated the importance of systematic prostate biopsy results in the risk assessment for newly diagnosed patients with prostate cancer. METHODS: We studied 1265 patients enrolled in CaPSURE (a longitudinal registry of patients with prostate cancer) who underwent radical prostatectomy as definitive local treatment of their prostate cancer. Preoperative characteristics, including age, race, prostate-specific antigen (PSA) level at diagnosis, clinical T stage, biopsy Gleason score, and percent positive prostate biopsies at the time of diagnosis, were determined for each patient. Disease recurrence was defined as PSA level of 0.2 ng/mL or greater on two consecutive occasions after radical prostatectomy or the occurrence of a second cancer treatment more than 6 months after surgery. Cox proportional regression analysis was performed to determine the significant independent predictors of disease recurrence. Patients were assigned to previously described risk groups on the basis of clinical tumor stage, PSA at diagnosis, and biopsy Gleason score. The likelihood of disease recurrence for each risk group, stratified according to the percentage of positive biopsies (0% to 33%, 34% to 66%, and more than 66%), was determined using the Kaplan-Meier method and compared using the log-rank test. RESULTS: The median follow-up was 3.3 years after surgery. The serum PSA level at diagnosis, biopsy Gleason score, percent positive biopsies, and ethnicity were significant independent predictors of disease recurrence. The percentage of positive prostate biopsies was a significant predictor of disease recurrence for low, intermediate, and high-risk patients. For patients with high-risk disease, the likelihood of disease recurrence 5 years after surgery was 24%, 34%, and 59% for patients with 0% to 33%, 34% to 66%, and more than 66% positive biopsies, respectively. CONCLUSIONS: Serum PSA, biopsy Gleason score, and percent positive biopsies were significant predictors of disease recurrence in this population. The percent positive biopsies may be useful in identifying high-risk patients suitable for definitive local therapy. PMID- 11927315 TI - Effect of radical prostatectomy on urethral blood flow. AB - OBJECTIVES: To study the effect of radical retropubic prostatectomy on urethral microcirculatory blood perfusion and urinary continence. Urinary stress incontinence after radical prostatectomy depends mainly on intrinsic distal sphincter integrity. METHODS: Urethral laser Doppler flowmetry was performed in 37 patients before and in 23 patients after (mean 15 months, range 2 to 23) radical prostatectomy for clinically localized prostate cancer. An endoscopic laser Doppler flow probe with a diameter of 1.8 mm was introduced through the working channel of a cystoscope and gently applied to the urethral mucosa at an intravesical pressure of 30 cm H(2)O. Blood flow was expressed simultaneously as a digital reading on the flowmeter (milliliters blood per minute per 100 gram tissue). The mean arterial blood pressure was monitored noninvasively. The Mann Whitney U test was used for statistical analysis. RESULTS: Membranous urethral blood flow significantly decreased after radical prostatectomy from 18.8 +/- 7.9 mL/min per 100 gram tissue to 11.9 +/- 7.7 mL/min per 100 gram tissue (P = 0.04). However, the difference in the blood flow did not statistically differentiate continent patients from patients with urinary stress incontinence (P = 0.47). No correlation between urethral microcirculation and mean arterial blood pressure could be detected. CONCLUSIONS: Retropubic radical prostatectomy decreases membranous urethral microcirculation. However, this loss of vascularization does not have a major effect on distal sphincter function and continence after radical prostatectomy. PMID- 11927316 TI - Treatment of Peyronie's disease by incomplete circumferential incision of the tunica albuginea and plaque with bovine pericardium graft. AB - OBJECTIVES: To study the surgical treatment of Peyronie's disease using a single incision in the tunica albuginea and fibrotic plaque and repair of the defect by bovine pericardium graft. METHODS: Between April 1999 and May 2001, 33 patients who had had symptoms of Peyronie's disease for more than 12 months and whose disease had been stable for more than 6 months underwent surgery. The reason for surgery was deformity of the penis that made sexual intercourse, even with erection, difficult or impossible. Two paraurethral incisions were made in Buck's fascia to separate the neurovascular bundle from the tunica albuginea. An incomplete circumferential incision, forked at the extremities, was made in the tunica albuginea and in the plaque at the point of maximal curvature, followed by a septal incision for the lengthening of the short side of the penis. A bovine pericardium graft was used to repair the defect in the tunica. All procedures were performed by a single surgeon (P.H.E.). The average follow-up time was 19.4 months (range 5 to 30). RESULTS: No rejection or retraction of the graft was observed. All patients maintained their state of preoperative erection, with the penis corrected in 87.9% of cases and with discrete curvature (less than 15 degrees ) in 12.1%. All recovered their ability to penetrate with no difficulty. In addition, an intraoperative average increase of 2.21 cm (range 1 to 4) occurred in the size of the penis. CONCLUSIONS: This procedure is effective for all types of penile deformity, regardless of the plaque characteristics. PMID- 11927317 TI - Novel assay for determining DNA organization in human spermatozoa: implications for male factor infertility. AB - OBJECTIVES: To present a novel assay for determining DNA organization in fertile human spermatozoa and establish morphometric parameters for these samples. The three-dimensional organization of DNA within a cell nucleus is intimately related to cellular function. For example, it has recently been demonstrated that normal sperm DNA organization may be necessary for successful in vitro fertilization in the mouse. METHODS: Semen from 12 fertile volunteers was tested for sperm DNA organization using our nuclear matrix stability assay. Sperm DNA then underwent computerized digital image analysis and standards of normal were established. RESULTS: Sperm DNA organization was constant in all samples tested. Normal parameters established included mean nuclear matrix diameter (9.17 +/- 1.59 microm), mean DNA halo diameter (20.56 +/- 2.53 microm), mean halo area (66.88 +/ 7.92 microm(2)), and mean nuclear matrix area (32.98 +/- 4.3 microm(2)). CONCLUSIONS: This assay may be used to determine DNA organization in a semen sample. Defining sperm DNA organization may be important clinically, because normal DNA organization is necessary for normal cellular function. PMID- 11927318 TI - Early nasogastric tube removal combined with metoclopramide after postchemotherapy retroperitoneal lymph node dissection for metastatic testicular nonseminomatous germ cell tumor. AB - OBJECTIVES: To evaluate the efficacy of combining early nasogastric tube (NGT) removal and metoclopramide after postchemotherapy retroperitoneal lymph node dissection (RPLND) in decreasing complications, time to diet tolerance, and discharge. METHODS: We performed a retrospective study of 31 patients who underwent postchemotherapy RPLND at the University of Texas M. D. Anderson Cancer Center between 1994 and 1996, who were treated with a clinical care pathway that included NGT removal on postoperative day 1 and immediate use of promotility agents such as metoclopramide. A comparison was made with 42 historic controls treated from 1988 to 1994 who were managed with an NGT until the return of flatus and minimal promotility agents. RESULTS: Both groups had comparable age, chemotherapy, and surgical procedures. The study group had fewer complications, with an earlier tolerance of solid food (median 5 +/- 2.5 days compared with 7 +/ 5.1 days in controls, P = 0.000) and discharge (median 6 +/- 2.6 days compared with 9 +/- 6.9 days in controls, P = 0.000). NGT replacement was required in 3.2% of study patients versus 9% of controls. Multiple regression analyses demonstrated that the number of NGT days was an independent predictor of early return of bowel function and length of hospitalization (P = 0.000), and metoclopramide was not. CONCLUSIONS: Early NGT removal combined with metoclopramide after postchemotherapy RPLND allowed earlier diet tolerance and shortened hospital stays without increased complications. The role that promotility agents play as a necessary component of safe, early NGT removal is unclear. PMID- 11927319 TI - Modified technique of dorsal plication for penile curvature with or without hypospadias. AB - OBJECTIVES: Since Nesbit reported dorsal plication at the lateral aspect of each corpus cavernosum to straighten the bending penis, dorsal plication on each side of the penis at the 2-o'clock and 10-o'clock positions has been a mainstay for correction of congenital or acquired penile curvature. However, because only the 12-o'clock position proved to be the nerve-free zone as determined by the histologic research, Baskin advocated that dorsal plication be done at the 12 o'clock position. METHODS: We performed plication at the 12-o'clock position of the corporeal body to repair penile curvature with and without hypospadias. We modified Baskin's plication technique, approximating the outer edges of the incisions made through the tunica albuginea. RESULTS: We repaired penile curvature with hypospadias in 10 patients and penile curvature without hypospadias in 3 patients, without complications. CONCLUSIONS: Penile curvature due to corporeal disproportion could be repaired by dorsal plication with incisions of the tunica at the 12-o'clock position, irrespective of the presence or absence of hypospadias. PMID- 11927321 TI - Outcomes analysis of 64 consecutive open pediatric renal and upper ureteral operations. AB - OBJECTIVES: To report a recent consecutive series of children undergoing open renal and upper ureteral surgery that was analyzed for outcomes, particularly morbidity and length of stay. METHODS: A total of 64 children underwent open renal or upper ureteral surgery using an upper abdominal retroperitoneal approach. RESULTS: The mean operation time was 206 minutes, with a mean estimated blood loss of 20 mL. The mean analgesic dosage during the in-house postoperative period was 2.9 doses. The mean length of stay was 21.3 hours. A single complication occurred. One patient had a postoperative temperature of 38.6 degrees C, which was attributed to a pulmonary causation. CONCLUSIONS: For now, open surgery remains the standard approach for pediatric renal and upper ureteral anomalies and diseases with an expected postoperative result and course similar to that of laparoscopically performed procedures. PMID- 11927323 TI - A novel laparoscopic specimen entrapment device to facilitate morcellation of large renal tumors. AB - A reusable laparoscopic instrument consisting of a flexible deployment ring and a barrel was fabricated, and an impermeable sac was sutured to the flexible ring before entrapment of the specimen and morcellation. The laparoscopic specimen entrapment device facilitated placement of large renal tumors within a sac for morcellation. PMID- 11927324 TI - Simultaneous multiple double pigtail stents for malignant ureteral obstruction. AB - Ureteral obstruction caused by extrinsic compression from pelvic malignancies often persists after ureteral stent placement. We describe a new technique and present our experience with the simultaneous placement of two ipsilateral ureteral stents after simple stenting has failed. PMID- 11927325 TI - Novel technique for combined repair of postirradiation vesicovaginal fistula and augmentation ileocystoplasty. AB - Concomitant vesicovaginal fistula and significant bladder contracture after radiation is an uncommon but complex urologic problem. We describe a surgical technique to address both issues and present our preliminary clinical data. PMID- 11927326 TI - A congenital scrotal mass consistent with an adnexal polyp of neonatal skin. PMID- 11927327 TI - Munchausen syndrome presenting as gross hematuria in two women. AB - Munchausen syndrome is an uncommon disorder in which patients present with fictitious disorders and a self-destructive urge to undergo invasive procedures. We present 2 cases of nurses who presented with recurrent urinary tract infections, flank pain, and gross hematuria. One patient had such severe hematuria as to require transfusions of more than 1000 U of packed red cells during a 30-year period. Both patients underwent extensive investigations--all of which were normal. One patient even underwent nephrectomy, which showed normal pathologic findings. Both were found to be phlebotomizing themselves and infusing blood into their bladders. PMID- 11927328 TI - Staghorn calculus endotoxin expression in sepsis. AB - Staghorn calculi are infrequent and generally are infected stones. Struvite or apatite calculi are embedded with gram-negative bacteria, which can produce endotoxin. Sepsis syndrome may occur after surgical therapy or endoscopic manipulation of infected or staghorn calculi. Sepsis, which can occur despite perioperative antibiotic use, may be due to bacteremia or endotoxemia. We present a child with an infected staghorn calculus who developed overwhelming sepsis and died after percutaneous stone manipulation. Endotoxin assay of stone fragments demonstrated an extremely high level of endotoxin despite low colony bacterial culture growth. This is the first reported case in which endotoxin was demonstrated in stone fragments from a child who died of severe sepsis syndrome after percutaneous staghorn stone manipulation. PMID- 11927329 TI - Anuria due to bilateral ureteral obstruction by Aspergillus flavus in an adult male. AB - Mycelial clumps or bezoars (fungus balls) as a cause of upper urinary tract obstruction are rare, with fewer than 60 cases previously reported. Anuria due to bilateral ureteral obstruction with mycelial clumps is most rare. We report a man with bilateral ureteral obstruction by Aspergillus flavus and describe his diagnosis and treatment. It is remarkable that imaging modalities, including excretory urography, computed tomography, and retrograde ureteropyelography, did not identify filling defects to suggest a fungal causation of the ureteral obstruction, even in the presence of a rapidly progressing process. For this reason, one should consider a fungal infestation in high-risk patients with obstructive uropathy of unknown etiology. PMID- 11927330 TI - Neonatal adrenal hemorrhage manifesting as acute scrotum: timely diagnosis prevents unnecessary surgery. AB - Neonatal adrenal hemorrhage presenting as scrotal swelling has been reported in 17 cases, with unnecessary surgical exploration in 7. We report 2 new cases, emphasizing the knowledge of this clinical association and the role of ultrasonography in the differential diagnosis for this specific condition and in all cases of neonatal acute scrotum. PMID- 11927332 TI - Primary signet ring cell carcinoma of the urinary bladder successfully treated with intra-arterial chemotherapy alone. AB - A 65-year-old man presented with the complaint of gross hematuria. Cystoscopy revealed a sessile tumor on the left bladder wall. It was diagnosed as primary signet ring cell carcinoma of the bladder (T3bN0M0). The patient did not want surgical treatment. Therefore, three courses of arterial infusion of carboplatin were administered at 3-week intervals. Complete remission was obtained and has been maintained for 44 months. Our case appears to be the first report of successful treatment with chemotherapy alone of an infiltrating signet ring cell carcinoma of the bladder. PMID- 11927331 TI - Urethral erosion of tension-free vaginal tape. AB - Tension-free vaginal tape (TVT) is gaining popularity as a treatment of choice for women with stress urinary incontinence. It is a minimally invasive procedure with reported short operative and postoperative hospitalization times and low complication rates. We describe urethral erosion of a TVT sling material in a 55 year-old woman who presented with immediate postoperative urinary retention. The sling material was surgically removed and the urethral defect repaired, with the patient continent at the 3-month follow-up visit. A mid-urethral synthetic sling such as the TVT can erode into the urethra. PMID- 11927333 TI - Large granular cell tumor of the penis in a 53-year-old man with coexisting prostate cancer. AB - Granular cell tumors are soft tissue neoplasms that rarely involve the male external genitalia. Thus far, only 7 cases of granular cell tumor of the penis have been reported. We report a case of granular cell tumor of the penis in a man undergoing radical retropubic prostatectomy for organ-confined adenocarcinoma of the prostate. PMID- 11927334 TI - Pelvic lipomatosis associated with proliferative cystitis occurring in two brothers. AB - Pelvic lipomatosis is a rare disorder of benign mature adipose tissue proliferation around the bladder and rectum. Most cases are associated with proliferative cystitis, in particular, cystitis glandularis. The etiology of pelvic lipomatosis and its association with proliferative cystitis are not well understood. This is the first reported case of familial pelvic lipomatosis. The pathogenesis and the possibility that this disorder could be secondary to genetic abnormalities of the HMG-IC (high mobility group) gene on chromosome 12 are discussed. PMID- 11927335 TI - Partial vesiculectomy in an infertile man with seminal vesicle cyst, ipsilateral renal agenesis, and cryptorchidism. AB - Congenital seminal vesicle cysts associated with renal agenesis are uncommon, but are currently detected more frequently with the use of sectional imaging procedures. Approximately 200 cases have been reported. The unique feature of our case is the combination of this disorder with an ipsilateral undescended testis. Our patient underwent partial vesiculectomy, in which the cyst was removed and the seminal vesicle remnant with its vas deferens was preserved. A review of the infertile cases and the impact of surgical treatment on fertility are discussed. Features that render partial vesiculectomy applicable and the potential effect of this procedure on fertility are highlighted. PMID- 11927336 TI - Nerve growth factor and chronic prostatitis/chronic pelvic pain syndrome. AB - OBJECTIVES: To investigate whether the pain experienced by patients with chronic prostatitis/chronic pelvic pain syndrome (CPPS) may be related to the expression of nerve growth factor (NGF), induced by inflammation and tissue injury experienced as a result of chronic inflammation. CPPS is a disease of unknown pathogenesis. METHODS: We measured the levels of NGF and the pro-inflammatory cytokine interleukin (IL)-6 and compared these with the levels of IL-8, interferon-gamma, IL-2, and IL-10 in the seminal plasma of 31 patients with CPPS and 14 controls using enzyme-linked immunosorbent assay technology. Results were correlated with health-related quality of life as measured by the multidimensional pain inventory, the McGill pain questionnaire, and the International Prostate Symptom Score. RESULTS: The cytokines analyzed were detectable in the seminal plasma from the patients with CPPS and controls. NGF correlated directly with pain severity (P <0.01) and IL-10 levels (P <0.04), and IL-6 correlated inversely with pain severity (P <0.03). CONCLUSIONS: These results suggest that NGF and cytokines that regulate inflammation (IL-6 and IL 10) may play a role in the pain symptoms experienced by patients with CPPS. PMID- 11927337 TI - Serum levels of human growth hormone during different penile conditions in the cavernous and systemic blood of healthy men and patients with erectile dysfunction. AB - OBJECTIVES: To detect changes in growth hormone (GH) serum levels during different penile conditions in the cavernous and systemic blood of patients with erectile dysfunction and compare them with the course of GH registered in healthy men. It has been suggested that human GH is involved in sexual maturation and plays a regulatory role in male reproductive function. Deficiency may result in fatigability, loss of sexual desire and erection, or oligospermia or azoospermia. It is assumed that the biologic effects of GH include insulin-like growth factor 1-mediated stimulation of endothelial nitric oxide formation. It has recently been demonstrated that GH serum levels in the systemic and cavernous blood of healthy men increases during developing penile erection. METHODS: Thirty-five healthy adult men and 45 patients with erectile dysfunction of either organogenic or psychogenic etiology were exposed to visual and tactile erotic stimuli to elicit penile tumescence and, in the group of healthy subjects, rigidity. Whole blood was simultaneously aspirated from the corpus cavernosum and the cubital vein during the different functional conditions of the penis. Serum levels of GH were determined by means of an immunoradiometric assay. RESULTS: In the healthy subjects, systemic GH serum levels significantly increased during penile tumescence, followed by a transient decline from tumescence to rigidity and detumescence. In the unselected patients, the mean GH levels during penile flaccidity were determined to be about sevenfold lower than those registered in the blood of the healthy men. During penile tumescence, the mean increase in the GH levels in the systemic and cavernous blood of psychogenic patients was comparable to that seen in healthy men, but, in the group of organogenic patients, this increase was found to be negligible. CONCLUSIONS: We believe our data provide strong evidence that GH may be of major importance in the maintenance of male erectile capability-probably through a stimulating effect on cyclic guanosine monophosphate generation in human cavernous smooth muscle-and that a decline in GH release may contribute to the manifestation of erectile dysfunction. PMID- 11927339 TI - Estrogen levels influence beta-3-adrenoceptor-mediated relaxation of the female rat detrusor muscle. AB - OBJECTIVES: To observe the expression of beta-3-adrenoceptor in the detrusor muscle in female rats and investigate the relaxant effect of beta-adrenoceptor agonists on detrusor muscle in ovariectomized rats with or without estrogen replacement therapy. METHODS: We first performed reverse transcriptase-polymerase chain reaction to demonstrate mRNA encoding the beta-3-adrenoceptor in the detrusor muscle from female rats. We then performed pharmacologic and physiologic studies to determine the effect of estrogen replacement therapy on the beta adrenoceptor-mediated relaxation of the detrusor muscle of the ovariectomized rats. RESULTS: Beta-3-adrenoceptor was expressed in the detrusor muscle in female rats with or without ovariectomy. A nonselective beta-adrenoceptor agonist relaxed precontracted detrusor muscle irrespective of ovariectomy or estrogen replacement in a dose-dependent manner; and a selective beta-3-adrenoceptor agonist relaxed the detrusor muscle more in ovariectomized rats than in ovariectomized rats with estrogen replacement or in control rats. CONCLUSIONS: Selective beta-3-adrenoceptor agonists relaxed the detrusor muscle of female rats with low estrogen levels. This result may give a clue to the treatment of frequent urination or incontinence in postmenopausal women who are not receiving hormonal replacement therapy. PMID- 11927338 TI - Clinical significance of p53, mdm2, and bcl-2 proteins in renal cell carcinoma. AB - OBJECTIVES: To improve our understanding of the clinical relevance of p53, mdm2, and bcl-2 protein overexpression in renal cell carcinoma, we retrospectively investigated the immunohistochemical expression of p53, murine double minute 2 (mdm2), and bcl-2 and the relationship of this expression to clinicopathologic characteristics. p53 regulates the transcription of downstream effectors such as the oncoprotein mdm2, and bcl-2 has been shown to inhibit apoptosis triggered by wild-type p53. METHODS: The expression of p53, mdm2, and bcl-2 protein was studied by immunohistochemical methods in paraffin-embedded nephrectomy specimens from 112 patients whose clinicopathologic data confirmed renal cell carcinoma. RESULTS: The expression of the p53 and bcl-2 protein was recognized in 15 (13.4%) and 52 (42.0%) cases, respectively; the expression of the mdm2 protein, however, was seen in only 2 cases (1.8%). No correlation was noted between these three proteins and any clinicopathologic parameters, except p53 expression and Stage T1 2/T3-4 (P = 0.0208). However, in multivariate analysis, stage (hazard ratio 3.586; P = 0.0002), expression of p53 (hazard ratio 6.090; P = 0.0126) and of mdm2 (hazard ratio 22.016; P = 0.0156), and coexpression of p53/mdm2 (hazard ratio 6.146; P = 0.0005) demonstrated a statistically significant effect on prognosis by proportional hazards regression tests. CONCLUSIONS: Our results indicate that stage, p53 expression, mdm2 expression, and coexpression of p53/mdm2 are useful to predict the clinical outcome in patients with renal cell carcinoma. PMID- 11927340 TI - John Syng Dorsey and post-colonial genitourinary surgery. PMID- 11927341 TI - Progression and survival in patients with T1G3 bladder tumors. PMID- 11927344 TI - Hand-assisted laparoscopic radical nephrectomy: comparison to open radical nephrectomy. PMID- 11927343 TI - Asymptomatic microscopic hematuria. PMID- 11927346 TI - Is vertical disparity used to determine azimuth? AB - The azimuth of a stimulus relative to the head can be determined from an extra retinal, eye-position signal plus an estimate of the retinal eccentricity of the image. Alternatively, azimuth could be determined from retinal-image information alone. Specifically, stimulus azimuth could be estimated from two derivatives of vertical disparity: vertical size ratio (which varies with azimuth), and the horizontal gradient of vertical size ratio (a measure of distance). Here we examine the determinants of perceived azimuth in viewing conditions that, theoretically, should favor the use of vertical disparity. We find no evidence that vertical disparity is used. Perceived azimuth was determined completely by felt eye position and the retinal eccentricity of the image. PMID- 11927347 TI - Disparity capture by flanking stimuli: a measure for the cooperative mechanism of stereopsis. AB - In this work the range and scaling properties of the cooperative (contextual) interaction that was first proposed by Julesz [Foundations of Cyclopean Perception, University of Chicago Press, Chicago, 1971] to address the correspondence problem in stereopsis is measured. To this end the effect that flanking difference of Gaussians (DoG) patches produce on a perception of a target pair of patches is studied. The relative depth configuration of the target pair can switch from the small disparity gradient to a large disparity gradient state as a result of cooperative effects of the flanking stimuli. It is found that the interaction strength falls with distance. Its range varies for different subjects from 2 to 3 DoG patch sizes and scales proportionally to the size of the stimuli. The results suggest that a very localized cooperative interaction is in effect at a broad range of spatial scales. PMID- 11927348 TI - Limitations on shape information provided by texture cues. AB - This paper uses visual, empirical and formal methods (Li & Zaidi, Vision Research, 40 (2000) 217; Li & Zaidi, Vision Research, 41 (22) (2001a) 2927) to examine the roles of oriented texture components in conveying veridical percepts of concave and convex surfaces that are pitched towards or away from the observer. The results show that pairs of components, oriented symmetrically around the axis of maximum curvature, combine to provide the geodesic orientation modulations that are critical for veridical shape perception. The degree of pitch determines the orientations of the critical pair of components. Perspective is crucial to the veridical perception of concavities and convexities, regardless of the degree of pitch. The results of this paper reconfirm that veridical shape perception depends on extracting critical patterns of oriented energy, but also show that the class of textures capable of conveying veridical percepts of developable shapes in general views is even more restricted than that identified by Li and Zaidi (Journal of Optical Society of America A, 18 (2001b), 2430). PMID- 11927349 TI - Generic and non-generic conditions for the perception of surface shape from texture. AB - Li and Zaidi (Vision Research 40 (2000) 217; 41 (2001) 1519) have recently argued that there are two necessary conditions for the perception of 3D shape from texture: (1) the texture pattern must have a disproportionate amount of energy along directions of principal curvature; and (2) the surface must be viewed with a noticeable amount of perspective. In the present article we present evidence that these conclusions are only valid under a limited set of non-generic viewing conditions. Other relevant factors that need to be considered in this context include the distribution of curvature on an object's surface and the set of possible viewing directions from which it can be observed. For generic viewing directions and patterns of curvature, the perception of surface curvature from texture is only minimally affected by the orientation spectrum of the texture pattern or the amount perspective in its optical projection. Li and Zaidi (Vision Research 41 (2001) 1519) have also identified two characteristic patterns of image contours, which they claim to be the only possible source of information within textured images for determining the direction of surface slant or the sign of surface curvature. In the present article we attempt to show that these characteristic patterns can only arise in natural vision for a limited set of non generic viewing directions. We also review several other factors that can influence the perceived direction of slant or the perceived sign of curvature, which have been identified previously by other investigators. PMID- 11927350 TI - On the delay in processing high spatial frequency visual information: reaction time and VEP latency study of the effect of local intensity of stimulation. AB - Saleh and Bonnet [Fechner Day 98, p. 344] have shown that, upon parafoveal stimulation and up to 6.5 c/deg, reaction time (RT) is a function of grating contrast multiplied by grating period. The present experiments extend these findings to foveal stimulation within a wider spatial-frequency (SF) range and to stimuli of different duration. Both RT and latency of visually evoked potentials (VEP) were measured. The findings might be explained by the following assumption: Most RT and VEP latency variations across the SF range are a result of local intensity factors (retinal contrast and width of grating bars). Residual RT variations were found that might be due to processing of high SFs by slower mechanisms than those processing low and medium SFs. PMID- 11927351 TI - Shape predominant effect in pattern recognition of geometric figures of rhesus monkey. AB - Three monkeys were trained successively with discrimination, concurrent matching to sample, and sameness-difference judgment tasks in which learning curves were compared. Then, the display duration for the stimuli was shortened to 100, 50, and 30 ms respectively to test the changes in accuracy and reaction time. All results in three experimental paradigms suggested consistently that the geometric shape (triangle, circle, and square) plays a more predominant role than topological features (the hole inside of a figure and the hole numbers) in monkey figure recognition. The results are different from the experiment by human subjects who presented hole predominant in figure recognition. Therefore, the precedence in perception depends on subject species, stimulus set, and ecological significance of the perceiving process. PMID- 11927352 TI - The hole precedence in face but not figure discrimination and its neuronal correlates. AB - The discriminative tasks of face and geometrical figure were trained in three rhesus monkeys. The hole feature speeds up learning of face discrimination, but hampers learning of figure discrimination. By reducing presentation duration of the stimuli, the detecting precedence appears to be the hole feature of the face component, while it appears to be shape feature of figures. The patterns of neuron firings in inferior temporal cortex (IT) are consistent with the context dependent precedence of hole feature. The results might suggest that the neural correlates exist not only in IT neurons, but also in combination with executive mechanism. PMID- 11927353 TI - Computing relief structure from motion with a distributed velocity and disparity representation. AB - Recent psychophysical experiments suggest that humans can recover only relief structure from motion (SFM); i.e., an object's 3D shape can only be determined up to a stretching transformation along the line of sight. Here we propose a physiologically plausible model for the computation of relief SFM, which is also applicable to the related problem of motion parallax. We assume that the perception of depth from motion is related to the firing of a subset of MT neurons tuned to both velocity and disparity. The model MT neurons are connected to each other laterally to form modulatory interactions. The overall connectivity is such that when a zero-disparity velocity pattern is fed into the system, the most responsive neurons are not those tuned to zero disparity, but instead are those having preferred disparities consistent with the relief structure of the velocity pattern. The model computes the correct relief structure under a wide range of parameters and can also reproduce the SFM illusions involving coaxial cylinders. It is consistent with the psychophysical observation that subjects with stereo impairment are also deficient in perceiving motion parallax, and with the physiological data that the responses of direction- and disparity-tuned MT cells covary with the perceived surface order of bistable SFM stimuli. PMID- 11927354 TI - Effects of inter-stimulus interval on perceived locations of successively flashed perisaccadic stimuli. AB - We investigated the perceived locations of two stimuli flashed successively near the time of saccade execution in a dark room. The inter-stimulus interval (ISI) between the flashes ranged from 80 to 240 ms. The results show that when the ISI was 120 ms or shorter, perceived locations of the flashes interacted with each other so that the perceived distance between them was equal to the distance between these flashes on the retina. When the ISI was 240 ms, this interaction was weak. These results suggest two hypotheses. Firstly, the relation of retinal locations of flashes is a strong cue for perceiving the flash locations when the ISI is shorter than about 120 ms.Secondly, the process of perceiving or memorizing a flash location requires some time. Therefore, the perceived location of the succeeding flash affects that of the preceding flash when the ISI as shorter than about 120 ms. PMID- 11927355 TI - The influence of stimulus chromaticity on the isoluminant motion-onset VEP. AB - Motion-onset visual evoked potentials (VEPs) were elicited by low spatial frequency chromatic isoluminant gratings presented in a central 7 degrees circular field. The chromatic composition of the stimuli was varied so as to modulate along different axes in colour space. For slow speeds (<5 degrees/s) changing the chromatic axis induced large response differences between the S- and L/M-cone VEPs. At faster speeds (5-12 degrees/s) the effects were not as marked. A dichotomy between the slow and fast responses was also shown to exist in terms of their contrast dependencies, the former exhibiting a stronger dependency on contrast than the latter. These findings suggest that neural substrates with chromatic sensitivity are involved in the generation of S- and L/M-cone mediated motion-onset VEPs at low velocities. At higher velocities, responses are generated by different mechanisms that possess little or no chromatic sensitivity. PMID- 11927357 TI - Impaired dodging in food-conflict following fimbria-fornix transection in rats: a novel hippocampal formation deficit. AB - It is well known that damage to the hippocampal formation (Ammon's horn, dentate gyrus, fimbria-fornix, and other pathways) produces impairments in spatial navigation and in certain forms of learning. Lesions within these structures have also been reported to produce some motor impairments, but the nature of these impairments is less understood. The present study examined the effects of fimbria fornix lesions on food wrenching and dodging, social interactions that occur when one rat attempts to steal food from a conspecific, who in turn attempts to protect the food by an evasive movement. Lesion effectiveness was confirmed histologically and electrophysiologically, by the loss of hippocampal rhythmical slow-wave activity (RSA or theta), and by changes in open field behavior (increased open field behavior, less thigmotaxis and more defecation). Analysis of the social interaction indicated when an eating control rat was approached by a conspecific that was attempting to steal its food, it prevented the theft by dodging, a rapid lateral maneuver involving forequarter turning and stepping with the rear limbs. Rats with fimbria-fornix lesions were significantly impaired in dodging and so were more likely to lose their food to the robber. This novel deficit in motor behavior is discussed in relation to contemporary theories of hippocampal function and it is suggested that the deficit may be caused by an inability of the fimbria-fornix damaged animals to disengage attention from eating in order to initiate an evasive movement to protect food. The finding of this novel deficit underscores the importance of considering both loss as well as release phenomena in the analysis of hippocampal formation function. PMID- 11927356 TI - Morphometric changes of the choriocapillaris and the choroidal vasculature in eyes with advanced glaucomatous changes. AB - PURPOSE: In addition to an elevated intraocular pressure a compromise of the ocular blood supply have been implicated in the pathogenesis of primary open angle glaucoma. The purpose of this study was to quantify morphologic changes in the choroid including choriocapillaris thickness and density and diameter of large choroidal vessels in post mortem eyes with advanced primary open-angle glaucoma. METHODS: We analyzed 20 eye bank eyes (provided from the Georgia Eye Bank in Atlanta) with end stage primary open-angle glaucoma and compared them with 20 age-matched control eye bank eyes. The eyes were processed for light microscopy and following variables were measured with a digital filar micrometer: density and diameter of large choroidal vessels in the macular and equatorial choroid; thickness of the choroid in the macular and equatorial region; density and thickness of choriocapillaris in the macular, peripapillary, and equatorial choroid; and peripapillary capillary-free area nasal and temporal to the optic disk. RESULTS: Eyes with glaucoma displayed a lower density of the capillaries of the choriocapillaris as compared to control eyes in the macular, temporal peripapillary, and equatorial choroid with 0.50-0.55 (p=0.018), 0.46-0.51 (p=0.016), and 0.50-0.55 (p=0.038), respectively. There was no significant difference for the choriocapillaris density in the nasal peripapillary choroid, the thickness of the capillaries of the choriocapillaris in all assessed locations, and the nasal and temporal peripapillary capillary-free zone of the choriocapillaris between eyes with glaucomatous damage and controls. Assessment of large choroidal vessels in the macular choroid showed that eyes with glaucoma had a decreased density of veins (11.7-38.9 mm(-2); p<0.001) and arteries (7.7 12.4 mm(-2); p=0.005) and arteries with a higher diameter (45.6-28.2 microm; p<0.001) as compared to control eyes. The large vessels in the equatorial choroid displayed no significant difference in diameter but a lower density (21.2-44.1 mm(-2); p=0.017) in eyes with glaucomatous damage as compared to controls. CONCLUSION: Eyes with advanced glaucomatous damage after long standing primary open-angle glaucoma exhibit several changes including decreased density of capillaries of the choriocapillaris and decreased density of large choroidal vessels. We cannot conclude from our study whether the observed vascular changes in the choroid are primary pathogenic factors or secondary phenomena. PMID- 11927358 TI - Effect of immobilization stress on brain polyamine levels in spontaneously hypertensive and Wistar-Kyoto rats. AB - The present study aimed to compare the basal brain polyamine levels and stress induced brain polyamine level changes in spontaneously hypertensive rats (SHR) and Wistar-Kyoto (WKY) rats. For immobilization stress, both strains underwent acute (3 h per day immobilization for 2 days), chronic (3 h per day immobilization for 15 consecutive days), or no immobilization stress (control group). Basal putrescine (PU) levels in frontal cortex and hippocampus of SHR (11.03 +/- 0.81 and 11.36 +/- 0.33 nmol/g tissue, respectively) were significantly higher than WKY rats (6.90 +/- 1.44 and 7.82 +/- 0.71 nmol/g tissue, respectively). However, there were no strain differences in basal spermidine and spermine levels between the two. After acute stress, the PU levels in frontal cortex and hippocampus (15.99 +/- 0.45 and 14.10 +/- 0.95 nmol/g tissue, respectively) were significantly increased in SHR as compared to the non stressed SHR (11.03 +/- 0.81 and 11.36 +/- 0.33 nmol/g tissue, respectively). In WKY rats, the PU level was significantly increased by acute stress in frontal cortex (11.68 +/- 1.12 nmol/g tissue) as compared to the non-stressed WKY (6.90 +/- 1.44 nmol/g tissue). After chronic stress, the PU levels in frontal cortex and hippocampus of SHR (12.44 +/- 0.54 and 11.34 +/- 0.66 nmol/g tissue, respectively) significantly decreased as compared to acute-stressed groups (15.99 +/- 0.45 and 14.01 +/- 0.95 nmol/g tissue, respectively). In WKY rats, after chronic stress, the PU level was significantly decreased in frontal cortex (5.73 +/- 0.36 nmol/g tissue) as compared to acute-stressed groups (11.68 +/- 1.12 nmol/g tissue). The PU levels in frontal cortex and hippocampus of acute-stressed (15.99 +/- 0.45 nmol/g tissue and 14.10 +/- 0.95 nmol/g tissue, respectively) and chronic-stressed (12.44 +/- 0.54 and 11.34 +/- 0.66 nmol/g tissue, respectively) SHR were significantly higher than acute-stressed (11.68 +/- 1.12 and 9.76 +/- 0.45 nmol/g tissue, respectively) and chronic-stressed (5.73 +/- 0.36 and 8.44 +/ 0.71 nmol/g tissue, respectively) WKY rats. The present study provides the higher basal PU level and stress-induced PU response in SHR as compared to WKY rats may be related to enhanced response of hypothalamic-pituitary-adrenocortical axis and sympathetic influence that may significantly contribute to the development of hypertension in SHR. PMID- 11927359 TI - Central noradrenergic blockade prevents autotomy in rat: implication for pharmacological prevention of postdenervation pain syndrome. AB - Following transection of peripheral nerve, rats exhibit autotomy, which is considered to be the animal model of postdenervation pain syndrome. It has been suggested that phantom limb pain is a result of peripheral denervation leading to reorganization of somatosensory pathways, particularly in the cerebral cortex, which is shown to depend upon central noradrenergic activity. In this study, sciatic and saphenous nerves were sectioned in the left hindpaw of 30 adult rats resulting in complete loss of pain sensation in the hindpaw. A group of rats received normal saline, compared to another group which received N-(2-) Chloroethyl-N-ethyl-2-bromobenzylamine (DSP4) injection 24 h prior to transection. The latter group was also compared to a third group whose central noradrenergic system were also blocked by bilateral injection of 6-OHDA into the ascending noradrenergic bundle 1 week prior to transection. A fourth group received contralateral cortical ablation in addition to peripheral nerve transection and was compared to the first group whose cortex remained intact. The animals were observed daily for 60 days and autotomy was scored in accordance to the system of Wall et al. After 1 week, control animals began to exhibit autotomy. In contrast, autotomy was absent in rats treated with DSP4, similar to rats which received 6-OHDA. Rats which had contralateral cortical ablation showed a considerably delayed onset of autotomy and a reduction in final autotomy scores. We conclude that autotomy, as a model of postdenervation pain syndrome, can be prevented by blockade of noradrenergically mediated cortical reorganization. The clinical implications of this finding are discussed. PMID- 11927360 TI - Transgenic mice expressing the human presenilin 1 gene demonstrate enhanced hippocampal reorganization following entorhinal cortex lesions. AB - We have examined the effects of the presence of the mutated human presenilin 1 gene (M146L; hps1*) on lesion-induced sprouting in the hippocampus of the mouse (C57/CBA). The entorhinal cortex was unilaterally lesioned with ibotenic acid in adult, male mice. Four weeks later the subsequent axonal sprouting in the dentate gyrus was analysed, by measuring the density of the synaptophysin immunocytochemical staining in the termination area of the entorhinal cortex axons. The data demonstrate that mice expressing either the human presenilin 1 gene (hps1) or the hps1* gene display a significantly increased density of immunocytochemical staining for synaptophysin, indicative of axonal sprouting, compared to the control mice. No (or a very small) sprouting response is observed in mice expressing the normal mouse ps1 gene. Taken together, these data indicate that the presence of a human ps1 gene, normal or with an Alzheimer's disease mutation, leads to enhanced plasticity in the mouse brain. PMID- 11927361 TI - Pregnancy alters lateral parabrachial nucleus but not hypothalamic Fos expression following hypotensive hemorrhage. AB - The goal of these experiments was to determine if hemorrhage-induced Fos expression in the hypothalamus and lateral parabrachial nucleus (LPBN) is altered by reproductive cycle phase or pregnancy. Conscious unrestrained female Sprague Dawley rats were subjected to a 16 ml/kg hemorrhage on the morning of the metestrus or proestrus phases of the estrous cycle, or on day 12-14 of pregnancy (mid-gestation). Hemorrhage induced a significant increase (p < 0.01) in the number of Fos-immunoreactive cell nuclei in the supraoptic nucleus, and in both the magnocellular and parvicellular components of the paraventricular hypothalamic nucleus, that did not differ between groups. In virgin females, hemorrhage also induced a significant increase in LPBN Fos expression that did not differ between metestrus and proestrus. In pregnant animals, there was an increase in basal LPBN Fos expression, but hemorrhage induced no further increase in the number of Fos-immunoreactive neurons in the LPBN. Mean arterial pressure decreased (p < 0.001) and plasma renin activity increased (p < 0.01) to a similar extent in all three groups after 16 ml/kg blood loss. In summary, the number of paraventricular and supraoptic nucleus neurons activated by hemorrhage is unaffected by estrous cycle phase or pregnancy. In contrast, pregnancy significantly attenuates the LPBN response to hemorrhage. PMID- 11927362 TI - Daily and circadian fluctuation in 2-deoxy[(14)C]-glucose uptake in circadian and visual system structures of the chick brain: effects of exogenous melatonin. AB - Previous studies show that several structures of the house sparrow visual system are metabolically rhythmic, as determined by 2-deoxy[(14)C]glucose (2DG) uptake, and that these metabolic rhythms depend upon rhythmic melatonin in this species. In many species of birds, high affinity binding of 2[(125)I] iodomelatonin is widespread in the brain, especially in visual system structures. The present study asks whether 2DG uptake is similarly rhythmic in the chick brain and whether exogenous melatonin administration affects 2DG uptake. Chicks were injected with 2DG and sacrificed 1 h later. Their brains were removed and processed for 2DG autoradiography. Chicks were injected during the late day with melatonin or saline prior to the 2DG injection and brain processing. We found that the visual suprachiasmatic nucleus showed both daily and circadian differences in 2DG uptake. Six of seven visual structures displayed daily uptake changes, while only two structures showed circadian fluctuations. Melatonin affected daytime 2DG uptake within visual suprachiasmatic nucleus and ectostriatum only. These results indicate that the chick circadian system is involved in the regulation of energy metabolism in the visual system but that the role for pineal melatonin in that regulatory process is a subtle one. PMID- 11927363 TI - Effects of intracerebroventricular insulin microinjection on renal sodium handling in kidney-denervated rats. AB - The role of the central nervous system (CNS) in the control of hydrosaline homeostasis has been strikingly demonstrated by several studies. Growing evidence suggests that insulin may exert an influence in the modulation of many brain functions. However, there are no available data examining the CNS effect of insulin injection on renal sodium handling. Also, to examine the influence of renal nerve activity during i.c.v. administration of insulin, unanesthetized, unrestrained rats were randomly assigned to one of nine separated groups: (a) sham-operated i.c.v. 0.15 M NaCl-injected (Co, pooled data, n = 37) and sham operated i.c.v. 0.42 ng. microl(-1) (n = 12), 4.2 ng.microl(-1) (n = 10) and 42.0 ng.microl(-1) (n = 11) insulin-injected rats (In); (b) renal-denervated i.c.v. 0.15 M NaCl (Co(Dx), n = 5), and insulin-injected rats (In(Dx), n = 5); and (c) subcutaneously insulin-injected rats (SC, n = 5). We showed that centrally administered insulin produced dose-related increased urinary output of sodium [Co: 855 +/- 85 Delta% min, 0.42 ng.microl(-1) In: 1189 +/- 308 Delta% min, 4.2 ng.microl(-1) In: 1461 +/- 594 Delta% min (p = 0.048), and 42.0 ng.microl(-1) In: 2055 +/- 411 Delta% min (p = 0.0001)], and dose-independently increased potassium excretion [Co: 460 +/- 28 Delta% min, 0.42 ng.microl(-1) In: 649 +/- 100 Delta% min (p = 0.016), 4.2 ng.microl(-1) In: 671 +/- 175 Delta% min (p = 0.003), and 42.0 ng.microl(-1) In: 669 +/- 70 Delta% min (p = 0.002)] compared to control. The urinary sodium excretion response to i.c.v. 42 ng.microl(-1) insulin injections were abolished by bilateral renal denervation. In addition, we showed that insulin-induced natriuresis occurred by increasing postproximal tubule sodium rejection (FEPP(Na)), and changed glomerular filtration rate (C(Cr)) at 42.0 ng.microl(-1) (p = 0.023) i.c.v. insulin microinjection but not at smaller insulin dose. The current data suggests that a blunted efferent insulin-sensitive nerve activity from periventricular region may contribute to the inability of renal tubules to handle the hydroelectrolyte balance. PMID- 11927364 TI - Chronic central infusion of orexin-A increases arterial pressure in rats. AB - We determined the cardiovascular responses as well as food and water intakes to chronic intracerebroventricular administration of orexin-A and orexin-B for 14 days in conscious rats. Chronic intracerebroventricular infusion of orexin-A (50 pmol/h) elicited a significant increase in systolic blood pressure on the third day (+15.6 +/- 2.9 mm Hg), and during the continuous intracerebroventricular infusion of orexin-A the blood pressure returned to the baseline levels at day 14. In contrast, chronic intracerebroventricular infusion of orexin-B (50 pmol/h) failed to change systolic blood pressure during the 14 days of experimental periods. Chronic intracerebroventricular infusions of neither orexin-A nor orexin B changed urinary catecholamine excretions, food and water intakes, and urine volumes at 7 and 14 days of infusion periods. Mean arterial pressure directly measured at 14 days did not differ among the groups of orexin-A, orexin-B, and artificial cerebrospinal fluid treatments. Both intravenous injections of pentolinium (5 mg/kg), a ganglion blocking agent, and CV-11974 (0.05 mg/kg), an AT(1) receptor antagonist, decreased arterial pressure; however, these responses were not different among the groups. These results suggest that central orexin-A participates in the short-term regulation of blood pressure; however, the contributions of central orexins to the long-term regulations of blood pressure, sympathetic nervous system, and appetite may be little. PMID- 11927365 TI - NMDA and AMPA/kainate glutamatergic agonists increase the extracellular concentrations of GABA in the prefrontal cortex of the freely moving rat: modulation by endogenous dopamine. AB - Using microdialysis in the prefrontal cortex, this study investigated first the effects of the ionotropic glutamatergic agonists NMDA and AMPA on extracellular concentrations of GABA, and second, the modulation of these effects by increasing endogenous dopamine. NMDA (20, 100, and 500 microM) and AMPA (1, 20, and 100 microM), perfused through the microdialysis probe for 60 min, produced a dose related increase of extracellular concentrations of GABA in the prefrontal cortex of the awake rat. NMDA 100 and 500 microM produced a maximal increase of extracellular GABA of 150 +/- 38% and 245 +/- 75% of baseline, respectively. AMPA 20 and 100 microM produced a maximal increase of extracellular GABA of 140 +/- 17% and 195 +/- 41% of baseline, respectively. NMDA and AMPA also increased extracellular concentrations of glutamate. Increases of extracellular GABA, and also of glutamate, produced by NMDA (500 microM) and AMPA (100 microM) were significantly blocked by the NMDA antagonist CPP (100 microM) and the AMPA/kainate antagonist DNQX (100 microM), respectively. To investigate whether dopamine modulates the increases of GABA produced by NMDA and AMPA, endogenous dopamine was increased with the dopamine uptake inhibitor nomifensine. Nomifensine (1, 100, and 1000 microM) produced a dose-related increase of dialysate dopamine (from 0.1 to 1.0 nM) but did not modify basal extracellular concentrations of GABA in the prefrontal cortex. However, increases of endogenous dopamine at 0.5-0.7 nM did potentiate the increases of extracellular GABA produced by AMPA (20 microM) (from 140% to 240% of baseline), but not by NMDA (100 microM), in this area of the brain. These effects were attenuated by the perfusion of (-)sulpiride (D2 antagonist), but not by the perfusion of SCH-23390 (D1 antagonist). These results suggest that glutamate, through the activation of both NMDA and AMPA/kainate ionotropic receptors, facilitates GABAergic transmission in the prefrontal cortex, and that dopamine can modulate the effects of glutamate through AMPA/kainate receptors on GABA transmission in this area of the brain. PMID- 11927366 TI - Fos induction in cortical interneurons during spontaneous wakefulness of rats in a familiar or enriched environment. AB - It has been repeatedly reported that Fos is spontaneously induced in several brain structures, including the cerebral cortex, during wakefulness. To ascertain whether cortical interneurons are involved in this state-dependent oscillation of gene regulation, we combined Fos immunocytochemistry with immunostaining of either parvalbumin or calbindin, known markers of cortical interneurons. Immunopositive neurons were examined in the sensorimotor and cingulate cortex. In rats perfused in basal conditions, a minor proportion (around 8%) of Fos immunoreactive neurons in the parietal cortex were also parvalbumin- or calbindin immunoreactive; these double immunostained cells accounted for 13% of the parvalbumin- and 34% of the calbindin-labeled neurons. Colocalization of Fos with either calcium-binding protein was instead not observed in the cingulate cortex. In rats stimulated by novel environmental cues during the period of wakefulness preceding perfusion, Fos-positive neurons increased markedly relative to unstimulated animals, and involved the majority of the calbindin- or parvalbumin labeled cell populations (60-75% and over 95%, respectively). In the neuronal populations in which Fos was induced by exposure to the enriched environment, the proportion of calbindin- and parvalbumin-labeled cells was larger than in the unstimulated cases, and the increment was statistically significant in the cingulate cortex. The results demonstrate that Fos induction occurring in the cortex during undisturbed wakefulness in a familiar environment involves a minor proportion of interneurons. Furthermore, the findings indicate that the addition of novel environmental stimuli results in an increase of Fos-expressing neurons whose recruitment, at least in the cingulate cortex, involves a higher proportion of interneurons than of projection neurons. PMID- 11927367 TI - Enhancement of paradoxical sleep by lights-off stimulation depends on sleep states. AB - This study was aimed at clarifying whether the lights-off stimulation effect on paradoxical sleep (PS) enhancement in albino rats depends on sleep and wakefulness states. Male Sprague-Dawley rat pairs were subjected to lights-off stimulation for 320 s when one of the two rats was in non-rapid-eye movement sleep (NREM) continuously for at least 64 s (NREM-off condition). Simultaneously, the other rat received the same lights-off stimulation during any sleep and wakefulness state (random-off condition). Each rat underwent both lights-off conditions. The sequence of the two lights-off conditions was counter-balanced. PS was enhanced during the lights-off period in both the NREM-off and random-off conditions. However, the increased amount of PS was greater in the NREM-off condition than the random-off condition. In the random-off condition, rats had more PS in the lights-off period than in the preceding and succeeding lights-on periods only if they were in high EEG amplitude NREM (HS) at lights-off. Thus, this study verified that the lights-off stimulation effect on PS enhancement in the albino rat depended on the sleep states. However, the lights-off treatment did not alter the relationship between PS and preceding NREM, or the hippocampal theta activity or the PS latency in the lights-off period. PMID- 11927368 TI - Modulation of apoptotic regulatory proteins and early activation of cytochrome C following systemic 3-nitropropionic acid administration. AB - The present study utilized the administration of the mitochondrial toxin, 3 nitropropionic acid (3-NP), to mimic the pathology associated with Huntington's disease (HD). 3-NP causes striatal cell degeneration via the inhibition of succinate dehydrogenase. There is growing evidence suggesting the role of apoptosis in 3-NP cell death. TUNEL staining, DNA fragmentation, and changes in bcl-2 mRNA levels have been associated with this metabolic impairment. We wish to further elucidate the apoptotic cascade in this model of HD pathogenesis. 3-NP was administered to rats intraperitoneally at 20mg/kg/day for up to 3 days. At 3 days, characteristic behavioral and morphological effects became evident. While cell death did not become apparent within the first 3 days, there were changes in the levels of apoptotic regulatory proteins and translocation of cytochrome c. Within 24 h after 3-NP administration there were elevations in both bcl-xl and bax. However, bcl-xl protein levels quickly returned to control levels while bax levels continued to increase, resulting in a detrimental bax/bcl-xl ratio. Bax has been demonstrated to facilitate cytochrome c release by forming mitochondrial pores. We saw cytochrome c translocate from the mitochondria to the cytosol approximately 24 h after initial 3-NP administration when compared to saline injected controls. This evidence generated using the 3-NP degeneration model may help elucidate the apoptotic cascade associated with HD neurodegeneration. PMID- 11927369 TI - Expression of c-fos gene activation during rough and tumble play in juvenile rats. AB - Rough and tumble (R&T) play is an intrinsic behavior in most mammals. However, unlike sex and aggression, play has not been well characterized in terms of neuronal circuitry. We employed in situ hybridization to explore the differences of c-fos mRNA activation in juvenile rats that had been allowed R&T play for a total of 30 min before sacrifice contrasted to animals with comparable histories that had received no play. Densitometric estimates of c-fos gene activation revealed that the deep and dorsolateral tectum, inferior colliculus, dorsal periaquaductal gray, ventromedial hypothalamus, dorsal and ventral striatum, and somatosensory cortex were significantly more activated in animals that had played than those that had not. Prior play dominance and amount of social experience had no clear effects on the levels of c-fos gene expression. This provides a variety of new hypotheses concerning the role of various brain areas in the elaboration of R&T play behavior, but the important role of other types of motor arousal in the differential effects were not evaluated in this study. PMID- 11927370 TI - Effects of 14-methoxymetopon, a potent opioid agonist, on the responses to the tail electric stimulation test and plus-maze activity in male rats: neuroendocrine correlates. AB - We have studied the effects of 14-methoxymetopon (HS 198), a potent opioid agonist, on the responses to the tail electric stimulation test and plus-maze activity of adult male rats. The prototype mu agonist morphine was used as the drug of reference. Besides we addressed the effects of HS 198 on the serum corticosterone levels and on serotonergic systems of discrete brain regions. Both drugs were administered subcutaneously. Morphine (5 mg/kg) and HS 198 (30 microg/kg) induced a similar effect on the nociceptive test, with both drugs significantly increasing the threshold for the vocalization afterdischarge, which is related to the emotional component of pain. In the plus-maze, morphine (5 mg/kg) and HS 198 (20 and 30 microg/kg) induced similar increases in the percentages of entries and time in the open arms, two parameters related to the anxiety state of the animals. The results indicate that HS 198 is far more potent than morphine in reducing the emotive/affective component of pain and in inducing an anxiolytic effect. HS 198 (30 microg/kg) also induced parallel increases in the serum corticosterone levels and the hypothalamic serotonin content. A possible correlation between the anxiolytic action of the drug and its effect on the hypothalamic serotonergic system is suggested. PMID- 11927371 TI - Dynamic changes and spatial correlation of EEG activities during cold pressor test in man. AB - To explore the effects of tonic cold pain in man, the pain rating (intensity and distress), skin temperature, and continuous EEG recording were conducted before, during, and after cold pressor test (CPT) in 15 young healthy males. The acquired electroencephalogram (EEG) data was analysed in four ways: (1) comparison of EEG topographic patterns and power spectra across baseline, CPT, and post-CPT; (2) dynamic EEG changes during CPT; (3) correlation of EEG activities at the isolated focal maxima across the three experimental stages; and (4) spatial correlation of EEG powers among the focal sites during CPT. Compared to baseline, CPT induced significant differences in EEG topographic patterns and power spectra, which showed the following characteristics. (A) The delta and theta activities increased in frontal areas with maxima at F8. (B) The alpha activities decreased in the posterior part of the head with maxima at POz. (C) The beta activities increased in the peripheral bi-temporal regions. (D) The decrease of alpha and increase of beta activities occurred immediately after the onset of CPT, but the increase of delta activity showed a relatively gradual process. (E) Individual consistency was significantly observed in delta power at F8 and alpha-1 power at POz across the stages. (F) Two independent spatial clusters of EEG activation, fronto-temporal delta-theta-beta activities and posterior parietal alpha activities emerged during CPT. This new evidence and the detailed EEG effects in CPT may enhance our understanding of the dynamics in cerebral processing of tonic noxious information. Alpha reduction may reflect the attention processing in nociceptive input, and the delta/theta/beta activation may be related to the motivational modulation of the brain. PMID- 11927372 TI - Effects of hypergravity on the morphological properties of the vestibular sensory epithelium. I. Long-term exposure of rats after full maturation of the labyrinths. AB - The effect of prolonged exposure to hypergravity on the morphology of vestibular epithelia of rats was investigated. At the age of 1 month, i.e., when vestibular end organs are fully maturated, three rats were transferred to a hypergravity environment of 2.5 g inside a large radius centrifuge. After 9 months, vestibular epithelia of these animals and of three control animals were immunohistochemically labeled for actin and tubulin. The apical cross-sectional area of epithelial cells of hypergravity exposed rats appeared to be smaller in all end organs. Area reduction was 1.9% in the saccule (not significant), 5.0% in the utricle (p < 0.005), and 11.6% in the crista (p<<0.001). No indications for a deterioration of vestibular functioning were observed. PMID- 11927373 TI - Effects of excitotoxic median raphe lesion on working memory deficits produced by the dorsal hippocampal muscarinic receptor blockade in the inhibitory avoidance in rats. AB - The experiments investigated the interactions between median raphe nucleus (MRN) serotonergic and septo-hippocampal muscarinic cholinergic systems in the modulation of forming and storing performances of working memory. Rats with ibotenic acid-induced MRN-lesion bilaterally received scopolamine (2-4 microg/each side) infusion into the dentate gyrus of the dorsal hippocampus and were tested in a single trial step-through inhibitory avoidance. Initial preference to the dark compartment (escape latency) was taken as the measure of non-mnemonic behaviours and response latency to enter the dark compartment immediately after the foot-shock was used to measure working memory. The high dose scopolamine infusion 10 min before the training decreased escape latencies in the sham-lesioned rats, whereas had no effect in the MRN-lesioned rats. Although MRN lesion per se did not alter response latency, it alleviated pre training scopolamine-induced decrease, but aggravated post-training scopolamine induced reduction in this parameter. These results suggest that the antagonistic interactive processes between MRN-serotonergic and hippocampal cholinergic systems modulate non-mnemonic component of working memory formation, whereas the storing performance of working memory is modulated by the synergistic interactions between these systems in the hippocampus, mainly in the dentate gyrus. PMID- 11927374 TI - The interaction of morphine and gamma-aminobutyric acid (GABA)ergic systems in anxiolytic behavior: using mu-opioid receptor knockout mice. AB - We investigated the interaction of morphine and gamma-aminobutyric acid (GABA)ergic systems in anxiolytic action using mu-opioid receptor knockout and wild-type female mice. An elevated plus-maze test was used to assess anxiolytic behavior. The anxiolytic activities were monitored after experimental animals were pretreated with morphine [15 mg/kg, subcutaneous (s.c.)] and 3 h later received a single dose of muscimol (0.5 mg/kg, s.c.). In another experiment, mice received a single dose of opioid antagonist, naloxone [1.0 mg/kg, intraperitoneal (i.p.)], or GABA(A) receptor antagonist, (+)bicucullin (2.0 mg/kg, i.p.), 2.5 h after the morphine and 30 min before the muscimol injection. Control mice received vehicle only. The results show that morphine enhanced muscimol-induced staying time in open arms by 160% in wild-type mice. Moreover, the effect of morphine in the wild type was inhibited by the pretreatment of either naloxone or (+)bicucullin. Autoradiographic analysis indicated that morphine-administration raised [(3)H]muscimol binding by around 25% of the basal level in posterior thalamic, mediodorsal thalamic, and amygdaloid areas, but not in the hippocampal area in wild-type mice. In contrast, morphine administration did not alter the [(3)H]muscimol binding in mu-opioid receptor knockout mice. The present results reveal that mu-opioid receptor may play a role in the modification of anxiolytic behavior regulated by GABAergic neurotransmission. PMID- 11927375 TI - [Phe(1)Psi(CH(2)-NH)Gly(2)]NC(1-13)NH(2) does not antagonize orphaninFQ/nociceptin-induced prolactin release. AB - The specificity of the orphaninFQ (OFQ)/nociceptin (N)-induced prolactin increase was determined in male and female rats by pretreating animals with different doses of [Phe(1)Psi(CH(2)-NH)Gly(2)]NC(1-13)NH(2), a compound originally reported to be a specific OFQ/N antagonist. In addition, the effect of naloxone pretreatment on OFQ/N-induced prolactin release was examined to determine if OFQ/N's effects were mediated by opiate receptors. Furthermore, dose response studies using [Phe(1)Psi(CH(2)-NH)Gly(2)]NC(1-13)NH(2) only were performed to determine potential agonist activity of this drug. Finally, growth hormone (GH) levels were determined as an index of specificity of the prolactin response. Our results confirm previous findings that OFQ/N potently stimulates prolactin release and that a gender difference exists in the magnitude of the response, with females showing a much greater response than male rats. The endocrine response is specific because OFQ/N potently stimulated prolactin, but not GH secretion. The prolactin response is not mediated by actions at opiate receptors because naloxone did not inhibit OFQ/N's effects on prolactin release. However, [Phe(1)Psi(CH(2)-NH) Gly2]NC(1-13) NH(2) did not antagonize OFQ/N's effects on prolactin release. Indeed, this drug acted as a potent agonist. Demonstrating pharmacological specificity of OFQ/N's effects on prolactin release awaits the development of more selective, specific antagonists. PMID- 11927376 TI - Parabrachio-cortical connections with the lateral hemisphere in the madagascan hedgehog tenrec: prominent projections to layer 1, weak projections from layer 6. AB - The present study was undertaken to further characterize and subdivide the rhinal cortex (insular and perirhinal areas) in the hedgehog tenrec (Echinops telfairi), a placental mammal with a rather low encephalisation index. Injections of wheat germ agglutinin-horseradish peroxidase into the dorsolateral pontine tegmentum revealed a prominent layer 1 projection to several rhinal target areas, while the rhinal cortex only stained weakly for the calcitonin gene-related peptide. Among the regions retrogradely labeled following tracer injections into the rhinal cortex, the parabrachial nucleus was considered the main origin of the tegmento cortical projection. This conclusion was based on the circumscribed pattern of termination, as well as the differences noted between the pattern of anterograde labeling and the pattern obtained by thyrosine hydroxylase immunohistochemistry. The tracer injections into the dorsolateral tegmentum also revealed numerous retrogradely labeled cells in the layer 5 of the dorsomedial frontal cortex. In contrast, the rhinal cortex only showed few labeled cells and most of these cells were located in the layer 6/7. A comparison with other species indicates that the tenrec's parabrachial nucleus gives rise to the most extensive cortical projections but receives the least prominent input from the lateral cerebral hemisphere. The layer 6/7 projection may be a common mammalian feature but it is overshadowed by the layer 5 projection in higher mammals. PMID- 11927378 TI - Central infusion of vasopressin in male rats accelerates extinction of conditioned taste avoidance induced by LiCl. AB - In shock avoidance tasks, extinction is prolonged when vasopressin is infused into the lateral ventricle after an acquisition session. Experiments were performed to determine whether a dose of vasopressin that does not induce conditioned taste avoidance (CTA) could prolong extinction of a LiCl-induced CTA when it is infused into the lateral ventricle of Sprague-Dawley male rats after acquisition. The first experiment was designed to determine whether infusion of vasopressin into the lateral ventricle would induce a CTA. Consumption of a sucrose solution was paired with infusion of vasopressin or saline, and even after two pairings, none of the vasopressin-treated rats showed decreases in sucrose consumption. Therefore, in the second experiment, this same dose of vasopressin was infused into the lateral ventricle 50 min after consumption of a sucrose solution was paired with an injection of LiCl. Vasopressin increased the rate of extinction of the LiCl-induced CTA. These results are the opposite of what has been found after peripheral administration of vasopressin before acquisition and/or extinction of a LiCl-induced CTA. Possible reasons for the difference in the direction of the effect on extinction include differential effects of vasopressin depending on the route of administration, the timing of injection, and the presence of aversive effects produced by the neuropeptide. PMID- 11927377 TI - Leptin affects the electrical activity of neurones in the hypothalamic supraoptic nucleus. AB - The present experiments were undertaken to examine whether leptin affects the electrical activity of neurones in the supraoptic nucleus (SON) by using brain slice preparation of male Wistar and obese Zucker rats. Bath application of leptin (10(-8) - 10(-12) M) induced mainly inhibitory response in SON neurones of Wistar rats, although a minority showed excitation. These effects were observed in both continuously and phasically active cells. The inhibitory effect of leptin still persisted in low Ca(2+), high Mg(2+) medium. Bath application of tolbutamide, which is known to inhibit ATP-sensitive potassium channel activity, did not reverse the inhibitory effect of leptin on SON neurones. The effect of bath application of leptin was also tested in SON neurones of obese Zucker rats. Although leptin still affected the electrical activity of some SON neurones of Zucker rats, the proportion of unaffected neurones was significantly higher than in Wistar rats. The results suggest that leptin may inhibit the secretion of both oxytocin and vasopressin by inhibiting the electrical activity of neurones in the SON via direct action. This inhibitory effect of leptin may be exerted through mechanisms other than activation of ATP-sensitive potassium channels. PMID- 11927379 TI - Signalling, cycling and desensitisation of gonadotrophin-releasing hormone receptors. AB - Sustained stimulation of G-protein-coupled receptors (GPCRs) typically causes receptor desensitisation, which is mediated by phosphorylation, often within the C-terminal tail of the receptor. The consequent binding of beta-arrestin not only prevents the receptor from activating its G protein (causing desensitisation), but can also target it for internalisation via clathrin-coated vesicles and can mediate signalling to proteins regulating endocytosis and mitogen-activated protein kinase (MAPK) cascades. GnRH acts via phospholipase C (PLC)-coupled GPCRs on pituitary gonadotrophs to stimulate a Ca(2+)-mediated increase in gonadotrophin secretion. The type I GnRH receptors (GnRH-Rs), found only in mammals, are unique in that they lack C-terminal tails and apparently do not undergo agonist-induced phosphorylation or bind beta-arrestin; they are therefore resistant to receptor desensitisation and internalise slowly. In contrast, the type II GnRH-Rs, found in numerous vertebrates, possess such tails and show rapid desensitisation and internalisation, with concomitant receptor phosphorylation (within the C-terminal tails) or binding of beta-arrestin, or both. The association with beta-arrestin may also be important for regulation of dynamin, a GTPase that controls separation of endosomes from the plasma membrane. Using recombinant adenovirus to express GnRH-Rs in Hela cells conditionally expressing a dominant negative mutant of dynamin (K44A), we have found that blockade of dynamin-dependent endocytosis inhibits internalisation of type II (xenopus) GnRH Rs but not type I (human) GnRH-Rs. In these cells, blockade of dynamin-dependent internalisation also inhibited GnRH-R-mediated MAPK activation, but this effect was not receptor specific and therefore not dependent upon dynamin-regulated GnRH R internalisation. Although type I GnRH-Rs do not desensitise, sustained activation of GnRH-Rs causes desensitisation of gonadotrophin secretion, and we have found that GnRH can cause down-regulation of inositol (1,4,5) trisphosphate receptors and desensitisation of Ca(2+) mobilisation in pituitary cells. The atypical resistance of the GnRH-R to desensitisation may underlie its atypical efficiency at provoking this downstream adaptive response. GnRH-Rs are also expressed in several extrapituitary sites, and these may mediate direct inhibition of proliferation of hormone-dependent cancer cells. Infection with type I GnRH-R-expressing adenovirus facilitated expression of high-affinity, PLC coupled GnRH-R in mammary and prostate cancer cells, and these mediated pronounced antiproliferative effects of receptor agonists. No such effect was seen in cells transfected with a type II GnRH-R, implying that it is mediated most efficiently by a non-desensitising receptor. Thus it appears that the mammalian GnRH-Rs have undergone a period of rapidly accelerated molecular evolution that is of functional relevance to GnRH-Rs in pituitary and extrapituitary sites. PMID- 11927380 TI - Gap junction proteins and cell-cell communication in the three functional zones of the adrenal gland. AB - Mouse and monkey adrenal glands were used to study the relationships between gap junction protein expression, intercellular communication and adrenal zonation. Dye communication patterns were determined by incubating freshly excised and hemisected adrenal glands in Lucifer yellow, a gap junction permeable fluorescent dye. Immunohistochemical techniques were used to localize adrenal gap junction proteins. The combination of these two techniques permitted the correlation of gap junction proteins with dye transfer and hormone responses in specialized regions of the adrenal cortex. Lucifer yellow dye communication was most pronounced in the inner glucocorticoid/androgen-producing regions (zona fasciculata/zona reticularis), but was virtually absent in the outer mainly mineralocorticoid-producing region (zona glomerulosa). This pattern of dye communication was coincident with immunohistochemical localization of the gap junction protein, alpha(1)Cx43. The variations in communication and alpha(1)Cx43 expression within the adrenal cortex are thought to be relevant to normal physiological regulation of the adrenal gland. PMID- 11927382 TI - Overexpression of vasopressin in the rat transgenic for the metallothionein vasopressin fusion gene. AB - Arginine vasopressin (AVP) is a major antidiuretic hormone, the overproduction of which causes diluting hyponatremia in humans and is called the syndrome of inappropriate antidiuresis (SIAD). To study physiological changes resulting from AVP overproduction and to develop an animal model of hyponatremia, the human AVP gene was expressed under the control of the metallothionein promoter in transgenic (Tg) rats. Analyses of AVP immunoreactivity (irAVP) in the tissues revealed that the transgene is expressed mainly in the central nervous system. Gel filtration showed that irAVP in the brain and plasma was properly processed AVP. AVP purified from the brains of both Tg and control rats also exerted equal bioactivity to generate cAMP in LLC-PK1 cells. The founder rats did not show any physical or anatomical abnormalities. Under basal conditions, Tg rats had high plasma AVP levels (Tg 13.8 +/- 2.5 pg/ml; control 2.7 +/- 1.2 pg/ml; n=6 in both groups; means +/- S.E.M.), decreased urine volume, and normal plasma [Na(+)]. Hypertonic saline injected i.p. did not affect AVP secretion in Tg rats. In response to a zinc-supplemented liquid diet, plasma AVP decreased in control rats, but increased in Tg rats (Tg 32.7 +/- 2.7 pg/ml; control 1.0+/-0.1 pg/ml; n=6), resulting in hyponatremia (Tg 135.2 +/- 2.5 mEq/l; control 140.8 +/- 0.4 mEq/l; n=6). To our knowledge, this is the first transgenic animal to show diluting hyponatremia. This transgenic rat may therefore provide a useful model in which to investigate various physiological alterations resulting from the oversecretion of AVP which involve SIAD, stress response, behavior, and blood pressure. PMID- 11927381 TI - Adaptation to sustained high plasma vasopressin in water and electrolyte homeostasis in the rat transgenic for the metallothionein-vasopressin fusion gene. AB - Prolonged exposure of tissues to a receptor agonist often leads to adaptive changes that limit the subsequent responsiveness of the tissue to the same agonist. Recently, we have generated rats transgenic for the metallothionein I human arginine vasopressin (AVP) fusion gene (Tg), which produced high plasma AVP with relatively preserved renal water excretion, suggesting that there might be adaptive mechanism(s) for maintaining water and electrolyte homeostasis against chronic AVP oversecretion from the earliest stage of life. In this study, to investigate whether down-regulation of AVP V2 receptor (V2R), which could possibly be caused by long-standing high plasma AVP, participates in this adaptive mechanism(s), non-peptidic V2R antagonist OPC31260 was administered to reverse the down-regulation, and water loading was performed after V2R antagonist treatment had been withdrawn. Additionally, to confirm the down-regulation, Northern blotting analysis for V2R mRNA was carried out. Tg rats showed slightly decreased urine volume and water intake with an equivalent plasma [Na(+)] level (Tg 140.4 +/- 0.6 mEq/l; control 139.3 +/- 0.6 mEq/l) under basal conditions. After water loading using a liquid diet containing zinc, which stimulates the promoter region in the transgene, the urine increase showed only limited suppression with a dramatically increased plasma AVP level and mild hyponatremia (135.8 +/- 1.8 mEq/l) in Tg rats. When diet containing OPC31260 had been provided for 4 days until the day before the start of water loading, antidiuresis and hyponatremia (125.4 +/- 1.mEq/l) were significantly potentiated. V2R mRNA expression in kidney was significantly less in Tg rats than in control rats under basal conditions, and this suppression was restored by OPC31260 treatment to levels comparable with those of control rats. These results suggest that long standing high plasma AVP causes V2R down-regulation, and it may play an important role in the adaptive mechanism(s) for maintaining water and electrolyte homeostasis in chronically AVP-overexpressing rats. PMID- 11927383 TI - Glucose-inducible hypertrophy and suppression of anion efflux in rat beta cells. AB - Hypertrophy of beta cells from obese fa/fa rats is associated with increased sensitivity to basal glucose. Exposure to glucose in culture distorts insulin secretion more in beta cells from large than small islets from fa/fa rats. The aim of the present study is to investigate whether increased beta cell volume is associated with both glucose hypersensitivity and altered activity of the glucose sensitive anion conductance. Beta cells from fa/fa rats had increased volume compared with those from lean rats after 24 h culture. Three-day exposure to 25 mM glucose in culture induced 10-15% hypertrophy in beta cells from lean rats and basal secretion from intact islets was increased tenfold. Estimates of ion channel activity were made from measurement of radiolabeled ion efflux. Taurine efflux, a marker of glucose-regulated anion channel activity, was reduced after high glucose exposure but no alterations in glucose-dependent K+ efflux were detected. The reverse hemolytic plaque assay was used to determine the contributions of the number of secreting cells (recruitment) versus secretion per cell in beta cells from enlarged (>250 microm diameter), intermediate (125-250 microm) and small (<125 microm) islets from lean and obese rats exposed to conditions mimicking hyperglycemia. After overnight culture, basal secretion was twofold greater from beta cells of large fa/fa islets compared with all other groups. Recruitment at low glucose was increased in all lean or fa/fa beta cells derived from >125 microm islets. When beta cells from small islets were exposed to supra-physiological glucose for 3 days, recruitment was increased at basal glucose and blunted at high glucose. Glucose exposure converts the recruitment profile of beta cells from small islets to resemble that of beta cells from large islets while inducing cellular hypertrophy and reduced anion conductance. However, hypertrophy alone did not predict functional characteristics of overnight-cultured beta cells from fa/fa rats. PMID- 11927384 TI - Evidence for a sustained increase in clonal beta-cell basal intracellular Ca2+ levels after incubation in the presence of newly diagnosed Type-1 diabetic patient sera. Possible role in serum-induced inhibition of insulin secretion. AB - We have previously reported that newly diagnosed Type-1 diabetic patient sera potently suppressed insulin secretion from a clonal rat pancreatic beta-cell line (BRIN BD11) but did not alter cell viability. Here, we report that apoptosis in BRIN BD11 cells incubated in various sera types (fetal calf serum (FCS), normal human serum and Type-1 diabetic patient) was virtually undetectable. Although low levels of necrosis were detected, these were not significantly different between cells incubated in sera from different sources. ATP levels were reduced by approximately 30% while nitrite production increased twofold from BRIN BD11 cells incubated for 24 h in the presence of Type-1 diabetic patient sera compared with normal human sera. Additionally, ATP levels were reduced by approximately 40% and DNA fragmentation increased by more than 20-fold in BRIN BD11 cells incubated in FCS in the presence of a pro-inflammatory cytokine cocktail (interleukin-1beta, tumour necrosis factor-alpha and interferon-gamma), compared with cells incubated in the absence of cytokines. Nitric oxide production from BRIN BD11 cells was markedly increased (up to 10-fold) irrespective of sera type when the cytokine cocktail was included in the incubation medium. Type-1 diabetic patient sera significantly (P<0.001) raised basal levels of intracellular free Ca(2+ )concentration ([Ca(2+)](i)) in BRIN BD11 cells after a 24-h incubation. The alteration in [Ca(2+)](i) concentration was complement dependent, as removal of the early complement components C1q and C3 resulted in a significant reduction (P<0.01) of sera-induced [Ca(2+)](i )changes. We propose that the mechanism of Type-1 diabetic patient sera-induced inhibition of insulin secretion from clonal beta-cells may involve complement-stimulated elevation of [Ca(2+)](i) which attenuates the nutrient-induced insulin secretory process possibly by desensitizing the cell to further changes in Ca(2+). PMID- 11927385 TI - Insulin resistance in adult rat offspring associated with maternal dietary fat and alcohol consumption. AB - Maternal diet during pregnancy has been reported to alter the offspring's ability to respond to a glucose challenge. The current studies report changes in basal and insulin-stimulated, in vitro glucose uptake in red (soleus) and white (extensor digitorum longus) muscle fiber types, as well as whole body insulin responsiveness of adult rat offspring associated with their mother's dietary fat and alcohol content during pregnancy. The offspring of Harlan-derived Sprague Dawley female rats, dosed during pregnancy with ethanol (ETOH) via a liquid diet (35% of calories as ETOH) with either 12% or 35% of calories as fat, were compared with offspring from litters whose mothers were pair-fed an isocaloric amount of the liquid diet without ETOH. Maternal access to the liquid diets was terminated on day 20 of the pregnancies (sperm plug=day 0). The offspring were surrogate fostered within 48 h of birth to mothers which had consumed commercial chow throughout their pregnancy. Following weaning at 21 days of age, the offspring consumed only commercial rat chow and they were examined over the next 14 months for changes in glucose homeostasis as a consequence of in utero exposure to maternal dietary fat and/or alcohol. The 35% maternal fat diet resulted in both in vivo and in vitro decreases in insulin sensitivity. Thus, compared with adults whose mother's diet contained 12% fat, significant, in vitro muscle and in vivo whole body insulin resistance (measured by hyperinsulinemic euglycemic clamping) was observed in adult rats whose mothers consumed 35% of dietary calories as fat. The addition of ethanol to the maternal 35% fat diet further reduced the offspring's red muscle tissues in vitro response to insulin, but did not affect whole body insulin sensitivity. Muscle basal and insulin stimulated receptor tyrosine kinase activity were significantly decreased (approximately -50%) by the 35% fat maternal diet but there was no compensatory increase in serum insulin or glucose levels. Based upon both in vivo and in vitro data, these studies suggested that in utero exposure to 35% fat has a sustained effect on the adult offspring's glucose uptake/insulin sensitivity and that the effect is paralleled, at least in part, by decreased insulin receptor tyrosine kinase activity. In utero ETOH exposure resulted in the loss of basal and insulin stimulated, in vitro glucose uptake in red muscle fibers but maternal dietary ETOH had no detectable effect on either in vivo insulin sensitivity or muscle tyrosine kinase activity. PMID- 11927386 TI - Acute effects of fatty acids on insulin secretion from rat and human islets of Langerhans. AB - Fatty acids have both stimulatory and inhibitory effects on insulin secretion. Long-term exposure to fatty acids results in impaired insulin secretion whilst acute exposure has generally been found to enhance insulin release. However, there are conflicting data in the literature as to the relative efficacy of various fatty acids and on the glucose dependency of the stimulatory effect. Moreover, there is little information on the responses of human islets in vitro to fatty acids. We have therefore studied the acute effects of a range of fatty acids on insulin secretion from rat and human islets of Langerhans at different glucose concentrations. Fatty acids (0.5 mM) acutely stimulated insulin release from rat islets of Langerhans in static incubations in a glucose-dependent manner. The greatest effect was seen at high glucose concentration (16.7 mM) and little or no response was elicited at 3.3 or 8.7 mM glucose. Long-chain fatty acids (palmitate and stearate) were more effective than medium-chain (octanoate). Saturated fatty acids (palmitate, stearate) were more effective than unsaturated (palmitoleate, linoleate, elaidate). Stimulation of insulin secretion by fatty acids was also studied in perifused rat islets. No effects were observed at 3.3 mM glucose but fatty acids markedly potentiated the effect of 16.7 mM glucose. The combination of fatty acid plus glucose was less effective when islets had been first challenged with glucose alone. The insulin secretory responses to fatty acids of human islets in static incubations were similar to those of rat islets. In order to examine whether the responses to glucose and to fatty acids could be varied independently we used an animal model in which lactating rats are fed a low-protein diet during early lactation. Islets from rats whose mothers had been malnourished during lactation were still able to respond effectively to fatty acids despite a lowered secretory response to glucose. These data emphasise the complex interrelationships between nutrients in the control of insulin release and support the view that fatty acids play an important role in glucose homeostasis during undernutrition. PMID- 11927387 TI - Increased insulin sensitivity and upregulation of insulin receptor, insulin receptor substrate (IRS)-1 and IRS-2 in liver of Ames dwarf mice. AB - In the present study we have used hypopituitary Ames dwarf mice, which lack GH, prolactin and TSH, to investigate the consequences of the deficiency of these hormones on glucose homeostasis and on the initial components of the insulin signal transduction pathway in the liver. Ames dwarf mice displayed hypersensitivity to insulin since they maintained lower fasting glucose concentrations (73% of control values), had significantly reduced amounts of insulin (58% of control values), and exhibited an increased hypoglycemic response to exogenous insulin. Probably as a result of reduced insulin production, Ames dwarf mice displayed intolerance to glucose. The insulin-stimulated phosphorylation of the insulin receptor (IR) tended to be increased in the liver of Ames dwarf mice, while IR receptor protein content was increased by 38%. Insulin-stimulated phosphorylation of insulin receptor substrate (IRS)-1 and IRS 2 was increased by 61 and 72% respectively, while IRS-1 and IRS-2 protein levels were increased by 76 and 95%. The insulin-stimulated association of the p85 regulatory subunit of phosphatidylinositol (PI) 3-kinase with IRS-1 was increased by 28%, but unaltered with IRS-2. Interestingly, while the insulin-stimulated phosphotyrosine-derived PI 3-kinase activity was not changed, insulin-stimulated protein kinase B activation was increased by 41% in this tissue. These alterations may account for the insulin hypersensitivity exhibited by these animals. The present findings in long-lived Ames dwarf mice add to the evidence that insulin signaling is importantly related to the regulation of aging and life span. PMID- 11927388 TI - Adrenopause or decline of serum adrenal androgens with age in women living at sea level or at high altitude. AB - The present study aimed to determine adrenopause or reduction of serum adrenal androgens with age at high altitude and at sea level. It was a cross-sectional study performed in 210 women resident at high altitude (4340 m) and 123 women living in Lima (150 m), aged 20-70 years. Fasting early morning blood samples were obtained. Serum dehydroepiandrosterone (DHEA), DHEA sulphate (DHEAS), androstenedione, testosterone and estradiol were measured by radioimmunoassay. Serum testosterone concentrations were greater in women living at high altitude than in those resident at sea level. Serum concentrations of DHEA, DHEAS and androstenedione were lower in women living at high altitude than in those living at sea level. The DHEAS/DHEA ratio was significantly greater, and the androstenedione/testosterone ratio was lower in samples from women living at high altitude. Among women older than 50 years of age, a greater decline in serum concentrations of DHEA was observed in those living at high altitude than in those living at sea level. Among women 60-70 years of age, serum concentrations of DHEA at high altitude were 46.9% of those in women of the same age living at sea level. Decay of DHEAS at sea level and at high altitude occurred from the age of 40 years. The decline was faster at high altitude than at sea level, and in women aged 60-70 years serum values of DHEAS at high altitude were 56% of those at sea level. In the same age group, serum concentrations of androstenedione among those native to high altitudes were 27.34% of the value at sea level. At sea level, serum testosterone concentrations did not change with age from 20 to 70 years. In women aged 20-39 years and 50-59 years, serum testosterone concentrations were greater at high altitude than at sea level (P<0.05). In those aged 60-70 years, the concentrations were similar in those living at sea level and at high altitude. At sea level and at high altitude, the serum testosterone/estradiol ratio increased with age (P<0.0034 and P<0.0001 respectively). This ratio increased at an earlier age among those living at high altitude (40-49 years) than among those living at sea level (50-59 years). Multivariate analysis showed that altitude (P<0.0001) and greater chronological age (P<0.001) were associated with lower serum DHEAS concentrations. DHEAS was related to chronological age (P<0.0001). Low serum androstenedione concentrations were related to living at high altitude at birth and greater chronological age (P<0.0001). In conclusion, adrenopause is attained earlier and is of greater magnitude at high altitude than at sea level. PMID- 11927389 TI - Stimulation of progesterone production in human granulosa-lutein cells by lipoproteins: evidence for cholesterol-independent actions of high-density lipoproteins. AB - Low-density lipoproteins (LDL) have been consistently reported to stimulate ovarian steroidogenesis, apparently by the provision of cholesterol as a steroidogenic substrate. Recent studies suggest that high-density lipoproteins (HDL) can also deliver cholesterol to support progesterone synthesis in human granulosa-lutein cells. Therefore, this study investigated the contributions of (i) cholesterol delivery, (ii) cyclic AMP and (iii) protein kinase C (PKC) in the steroidogenic responses of human granulosa-lutein cells to HDL and LDL. Over a 24 h treatment incubation, HDL stimulated a larger increase in progesterone output than did LDL at equivalent cholesterol concentrations. Moreover, at equal protein concentrations (100 microg protein/ml), HDL doubled progesterone production by cells co-treated with a maximally effective concentration of 22R hydroxycholesterol, whereas LDL had no effect on the progesterone response to this membrane-permeable sterol. These observations indicate that the progesterone response to HDL is not solely due to the delivery of cholesterol as a steroidogenic substrate. Over 24 h, the stimulation of progesterone synthesis by HDL was additive with the response to a maximally effective concentration of dibutyryl-cAMP, but was unaffected by the down-regulation of PKC activity (by chronic pre-treatment with a tumour-promoting phorbol ester). We have concluded that HDL appears to stimulate progesterone production in human granulosa-lutein cells by a mechanism not solely reliant on cholesterol delivery. PMID- 11927390 TI - Influence of sex and gonadal status of sheep on cortisol secretion in response to ACTH and on cortisol and LH secretion in response to stress: importance of different stressors. AB - There are sex differences in the response to stress and in the influence of stress on reproduction which may be due to gonadal steroids but the nature of these differences and the role of the gonads are not understood. We tested the hypotheses that sex and the presence/absence of gonads (gonadal status) will influence the cortisol response to injection of ACTH, insulin-induced hypoglycaemia and isolation/restraint stress, and that sex and gonadal status will influence the secretion of LH in response to isolation/restraint stress. Four groups of sheep were used in each of three experiments: gonad-intact rams, gonadectomised rams, gonad-intact ewes in the mid-luteal phase of the oestrous cycle and gonadectomised ewes. In Experiment 1 (n=4/group), jugular blood samples were collected every 10 min for 6 h; after 3 h, two animals in each group were injected (i.v.) with ACTH and the remaining two animals were injected (i.v.) with saline. Treatments were reversed 5 days later so that every animal received both treatments. Experiment 2 (n=4/group) used a similar schedule except that insulin was injected (i.v.) instead of ACTH. In Experiment 3 (n=5/group), blood samples were collected every 10 min for 16 h on a control day and again 2 weeks later when, after 8 h of sampling, all sheep were isolated and restrained for 8 h. Plasma cortisol was significantly (P<0.05) elevated following injection of ACTH or insulin and during isolation/restraint stress. There were no significant differences between the sexes in the cortisol response to ACTH. Rams had a greater (P<0.05) cortisol response to insulin-induced hypoglycaemia than ewes while ewes had a greater (P<0.05) cortisol response to isolation/restraint stress than rams. There was no effect of gonadal status on these parameters. Plasma LH was suppressed (P<0.05) in gonadectomised animals during isolation/restraint stress but was not affected in gonad-intact animals, and there were no differences between the sexes. Our results show that the sex that has the greater cortisol response to a stressor depends on the stressor imposed and that these sex differences are likely to be at the level of the hypothalamo-pituitary unit rather than at the adrenal gland. Since there was a sex difference in the cortisol response to isolation/restraint, the lack of a sex difference in the response of LH to this stress suggests that glucocorticoids are unlikely to be a major mediator of the stress-induced suppression of LH secretion. PMID- 11927391 TI - Adrenomedullin modulates the neurohumoral response to acute volume loading in normal conscious sheep. AB - The physiological role of adrenomedullin (ADM) in volume and pressure homeostasis remains unclear. Accordingly, we assessed possible modulatory actions of ADM infusions on the neurohumoral response to acute volume loading with dextran in normal conscious sheep. Dextran (15 ml/kg), given with concurrent ADM (5.5 pmol/kg per min--raising plasma ADM from below detection to approximately 10 pmol/l) or vehicle control infusions, induced matched significant (P<0.001 by ANOVA) falls in hematocrit (27-30%) during both ADM and control and similar increases in right atrial pressure (approximately 10 mmHg). Compared with control, both systemic (P=0.033) and pulmonary (P=0.005) arterial pressure and peripheral resistance (P=0.004) were reduced during ADM but were raised post infusion. The dextran-induced increase in cardiac output was augmented by ADM (P=0.048). Dextran-induced increases in plasma atrial natriuretic peptide (ANP; P=0.008), brain natriuretic peptide (BNP; P=NS) and cyclic guanosine monophosphate (cGMP; P=0.003) were augmented post-ADM infusions. The dextran induced fall in plasma renin activity (PRA) was attenuated by ADM (P=0.039) whereas plasma aldosterone levels were unaltered. ADM augmented the increase in urinary volume during the second 2-h clearance period post-dextran. Our data indicate that ADM modifies the hemodynamic and hormonal response to an acute volume challenge, enhances natriuretic peptide secretion and reduces systemic vascular resistance. These results provide further evidence that ADM plays a physiological role in volume and pressure homeostasis. PMID- 11927392 TI - The effect of maternal undernutrition on ovine fetal growth. AB - Modifications in maternal nutrition during pregnancy can significantly disrupt fetal growth and subsequent post-natal health and survival. This study investigated the effects of undernutrition on fetal growth and the potential mechanisms involved. Tissue from pregnant ewes (n=27) was investigated on days 45, 90 and 135 of gestation (term = approximately 150 days). The thoracic girth (P<0.05) was greater in fetuses from nutrient restricted ewes on day 45 and there was also a trend towards an increased gut weight (P<0.08). By day 90, the fetal brain and thymus weight were lighter in underfed than in well-fed animals whilst the weight of the fetal ovaries was heavier (P<0.05). On day 135 the fetal heart, pancreas, thymus, gut and kidney weights were lighter in undernourished ewes (P<0.05). When expressed as a percentage of fetal body weight, significance was retained in the heart, pancreas and thymus (P<0.05). Bone growth was also affected. At day 90 the fetal femur and metatarsal were longer in underfed mothers (P<0.05). In contrast, the fetal humerus and scapula were shorter in underfed than in well-fed animals on day 135 (P<0.05) when the weight of the semitendinosus muscle (P<0.05) was also reduced. The fall in fetal glucose (P<0.1), insulin (P<0.01) and IGF-I (P<0.01) levels in underfed ewes on day 135 may have compromised fetal growth. Fetal plasma IGF binding protein-2 also increased between days 90 and 135 in underfed ewes (P<0.03), whilst levels were unaltered in well-fed animals. Although maternal and fetal plasma IGF-I levels increased with gestation (P<0.01) and the placentome morphology altered in all ewes (P<0.05), the fall in placental mass (P<0.05), amniotic and allantoic glucose concentrations (P<0.05) and maternal plasma glucose and insulin levels (P<0.05) in underfed ewes in late gestation may have compromised fetal substrate delivery. These perturbations in fetal development may have significant implications on adult health and carcass conformation, raising important health and economic issues in medical and agricultural sectors. PMID- 11927393 TI - Effects of thyroid hormones on pulmonary and renal angiotensin-converting enzyme concentrations in fetal sheep near term. AB - In the sheep fetus, pulmonary and renal concentrations of angiotensin-converting enzyme (ACE) increase towards term in parallel with the prepartum surges in plasma cortisol and tri-iodothyronine (T(3)). The ontogenic change in pulmonary ACE has been shown to be induced, at least in part, by cortisol but the role of the thyroid hormones is unknown. Therefore, this study investigated the effects of thyroid hormones on tissue ACE concentration in fetal sheep during late gestation. Pulmonary and renal ACE concentrations were measured in sheep fetuses after experimental manipulation of thyroid hormone status by fetal thyroidectomy and exogenous hormone infusion. In intact fetuses, pulmonary and renal ACE concentrations increased between 127-132 and 142-145 days of gestation (term 145 +/- 2 days), coincident with the prepartum rises in plasma cortisol and T(3). The ontogenic increment in pulmonary ACE concentration was abolished when the prepartum surge in T(3), but not cortisol, was prevented by fetal thyroidectomy. At 143-145 days, ACE concentration in the lungs and kidneys of the thyroidectomised fetuses were both lower than those in the intact fetuses. In intact fetuses at 127-132 days, pulmonary ACE was upregulated by intravenous infusions of either cortisol (2-3 mg/kg per day) or T(3) (8-12 microg/kg per day) for 5 days. Renal ACE was unaffected by cortisol or T(3) infusion. Therefore, thyroid hormones have an important role in the developmental control of pulmonary and renal ACE concentration in the sheep fetus towards term. In addition, the prepartum rise in plasma T(3) appears to mediate, in part, the maturational effect of cortisol on pulmonary ACE concentration. PMID- 11927394 TI - Perinatal growth and plasma GH profiles in adolescent and adult sheep. AB - Poor prenatal growth is associated with limited evidence of GH deficiency in adult humans, which may contribute to their increased risk of cardiovascular and metabolic disease. We therefore examined the effects of placental restriction of fetal growth (PR) on size at birth, neonatal fractional growth rate (FGR) and the circulating GH profile in adolescent and young adult sheep of both sexes. Moderate or severe PR decreased birth size and increased neonatal FGR of weight, crown-rump length and abdominal circumference. In adolescent males, mean and baseline GH concentrations correlated negatively and independently with birth weight and FGR of weight, and mean GH concentrations correlated negatively with current weight. In young adult males, mean GH concentrations correlated negatively and independently with birth shoulder height and FGR of shoulder height whilst, in young adult females, these correlations were positive. This suggests that restricted fetal growth and reduced neonatal growth rate in sheep are followed by elevated circulating GH in adolescent and adult males, but GH deficiency or increased GH clearance in adult females. PMID- 11927395 TI - Pit-1/GHF-1 and GH expression in the MCF-7 human breast adenocarcinoma cell line. AB - GH expression in mammary tumors has been related to the increase and spreading of cell proliferation. Using the MCF-7 human breast adenocarcinoma cell line, it has been demonstrated that autocrine GH-stimulated mammary carcinoma cell proliferation decreased the apoptosis rate and enhanced cell spreading. Surprisingly, no data are available about the presence of Pit-1 (the main pituitary regulator of GH) or GH expression in this cell line. Using RT-PCR, Western blot and immunohistochemistry, we have demonstrated the presence of both mRNA coding Pit-1 and GH as well as Pit-1 and GH protein in the MCF-7 cell line. These data could imply that Pit-1 may be an adequate target to inhibit breast cell proliferation. PMID- 11927396 TI - Atrial natriuretic factor and C-type natriuretic peptide induce retraction of human thyrocytes in monolayer culture via guanylyl cyclase receptors. AB - The natriuretic peptides signal through three receptor subtypes, of which two (NPR-A and NPR-B) are membrane-bound guanylyl cyclases for which the principal ligands are respectively atrial natriuretic factor (ANF) and C-type natriuretic peptide (CNP). In the human thyroid cell, a third receptor, NPR-C, has been implicated in the regulation of thyroglobulin, but functional roles for NPR-A and NPR-B have not yet been defined. In the present study we used RT-PCR to identify transcripts of all three receptor subtypes, both in human thyroid and in HTU-5 cells, a long-term culture of thyroid-derived cells. Both ANF and CNP induced a twofold increase in intracellular cGMP content in HTU-5 cells. Morphologic changes (a significant increase in cells of the retracted phenotype) were observed in ANF- and CNP-treated cells within 3 and 5 h of treatment respectively. Significant increases in retracted cell number were induced by ANF and CNP, but not the NPR-C-specific ring-deleted ANF analog, C-ANF(4-23), during a 15-day treatment. All three natriuretic peptides, however, induced a small (15 20%) but significant (P<0 small middle dot001) increase in DNA content per well. The stable analog of cGMP, 8-bromo-cGMP (8-BrcGMP; 1 mM), also increased the number of retracted HTU-5 cells, and was equipotent with the cAMP analog, 8 BrcAMP, in this effect. The cGMP-dependent protein kinase inhibitor, KT5823, however, had no significant effect on the ANF-induced increase in numbers of retracted cells. These results suggest that the actions of NPR-A and NPR-B, functional receptors in the human thyroid cell, may in part be mediated by cGMP induced alterations in the cytoskeleton. PMID- 11927397 TI - Nitric oxide is involved in interleukin-1alpha-induced cytotoxicity in polarised human thyrocytes. AB - Nitric oxide (NO) is a well-known mediator of autoimmune processes. In the thyroid gland, it is produced in response to interleukin 1 (IL-1) and may mediate cytokine action at an early stage of autoimmune thyroiditis. In this study, we have investigated whether NO is involved in cytokine-induced cytotoxic effects and epithelial barrier alterations in thyrocytes. Human thyroid epithelial cells were cultured as tight polarised monolayers on a permeable support and exposed or not to IL-1alpha (100 U/ml), alone or in combination with interferon-gamma (IFN gamma; 100 U/ml) added to the basal compartment. NO production was not detected in control thyrocytes, but was significantly induced by the combination of IL 1alpha with IFN-gamma, in a time-dependent fashion. Similarly, expression of the inducible isoform of nitric oxide synthase (NOSII), determined by immunoblot and immunofluorescence confocal microscopy, was not detected in control cells, but was markedly induced after 48-h exposure to both cytokines. This treatment significantly increased the release of cytosolic lactate dehydrogenase (LDH) in the apical and basolateral media and decreased transepithelial electrical resistance. Although IFN-gamma was not sufficient to induce NO production, it could by itself decrease transepithelial resistance and synergised the IL-1alpha effect on LDH release. The NOS inhibitor, L-nitro-arginine-methyl ester, suppressed the cytokine-induced NO production and decreased the LDH release, but failed to prevent the loss of transepithelial resistance. These results indicated that human thyrocytes express NOSII and produce NO in response to IL-1alpha+IFN gamma and suggest that NO acts as a mediator of cytokine-induced cytotoxicity in the thyroid gland and may promote the exposure of autoantigens to the immune system. In contrast, NO does not appear to mediate the cytokine-induced disruption of the thyroid epithelial barrier. PMID- 11927398 TI - Adrenergic regulation of catecholamine secretion from trout (Oncorhynchus mykiss) chromaffin cells. AB - The interaction between extracellular catecholamines and catecholamine secretion from chromaffin cells was assessed in rainbow trout (Oncorhynchus mykiss) using an in situ saline-perfused posterior cardinal vein preparation. This was accomplished by comparing the effects of adrenergic receptor agonists and antagonists on stimulus-evoked secretion. An acute bolus injection or extended perfusion with saline containing high levels of either noradrenaline or adrenaline did not affect the baseline secretion of catecholamines. However, catecholamine secretion in response to a bolus injection of the general cholinergic receptor agonist carbachol or electrical stimulation of the nerves innervating the chromaffin cells was abolished or reduced respectively, in preparations perfused with saline containing either catecholamine. To characterize the catecholaminergic inhibition of catecholamine release, secretion in response to carbachol and electrical stimulation was compared in preparations perfused with the adrenergic receptor agonists dobutamine (beta(1)), salbutamol (beta(2)), phenylephrine (alpha(1)) or clonidine (alpha(2)). Prior treatment with dobutamine or phenylephrine was without effect on baseline catecholamine secretion or stimulus-evoked secretion. In contrast, pre-treatment with salbutamol significantly inhibited catecholamine secretion in response to carbachol or electrical stimulation. Pre-treatment with clonidine did not affect carbachol-evoked secretion but did reduce catecholamine secretion during electrical stimulation. The significance of this adrenergic mechanism of regulating stimulus-evoked catecholamine secretion was further established using the adrenergic receptor antagonists nadolol (beta) or phentolamine (alpha). Catecholamine secretion in response to cholinergic stimulation was significantly enhanced in preparations perfused with saline containing nadolol. Furthermore, pre-treatment with phentolamine significantly enhanced adrenaline secretion in response to neuronal stimulation. These results suggest that the mechanisms of adrenergic inhibition of catecholamine secretion from trout chromaffin cells include activation of chromaffin cell membrane beta(2)-receptors and presynaptic alpha(2)-adrenergic receptors. PMID- 11927399 TI - Effects of dibutyryl cAMP on stanniocalcin and stanniocalcin-related protein mRNA expression in neuroblastoma cells. AB - Stanniocalcin is a polypeptide hormone that was first reported in fish as a regulator of mineral metabolism. Its recent identification in mammals has opened a new area of investigation in basic and clinical endocrinology. In the present study, regulation of the stanniocalcin (STC) and stanniocalcin related protein (STCrP) genes were investigated in mouse neuroblastoma cells (Neuro-2A) in relation to neuronal cell differentiation. Neuro-2A is an undifferentiated cell line that contains measurable levels of STCrP mRNA, but undetectable levels of STC mRNA. Treatment of the cells with either dbcAMP (1-4 mM) or 50 microM euxanthone (PW1) resulted in extensive differentiation and neurite outgrowth. However, only neurites of dbcAMP-treated cells developed varicosities, a phenotypic marker of axon formation. Furthermore, following differentiation induced by dbcAMP, there was an upregulation of STC and downregulation of STCrP mRNA levels. In the first 24 and 48 h of treatments, there was a maximum twofold induction and 1.5-fold reduction in STC and STCrP mRNAs respectively. Following 96 h of treatment, an additional 14-fold STC induction and 1.2-fold STCrP reduction were observed. The increase in STC mRNA levels was accompanied by a concomitant increase in axon-specific low molecular form microtubule-associated protein (MAP-2c) mRNA and varicosities on the neurites, suggesting a possible role for STC in axonogenesis. There was no induction of STC mRNA levels when PW1 was added into the culture media, whereas ionomycin (1-10 microM) had no observable effects on cell differentiation or STC/STCrP mRNA. Immunocytochemical staining of dbcAMP-treated cells revealed abundant levels of immunoreactive STC, particularly in the varicosities, with only weak staining in control, untreated cells. Antisense oligodeoxynucleotides transfection studies indicated that the expression of STC was a cause of varicosity formation and a consequence of cell differentiation. Our findings lend further support to the notion that STC is involved in the process of neural differentiation. PMID- 11927400 TI - LASIK and corneal ectasia. PMID- 11927402 TI - Patient perspectives on macular hole surgery. PMID- 11927404 TI - LASIK and vitreous pathology after LASIK. PMID- 11927406 TI - Posterior capsule opacification. PMID- 11927408 TI - Posterior capsule opacification. PMID- 11927412 TI - Incidence of NAION with cataract extraction. PMID- 11927409 TI - Bimatoprost vs. timolol. PMID- 11927413 TI - Incidence of NAION with cataract extraction. PMID- 11927414 TI - Correction of irregular astigmatism. PMID- 11927417 TI - Increasing geriatrics expertise in ophthalmology. PMID- 11927418 TI - Conductive keratoplasty for the correction of low to moderate hyperopia: 1-year results on the first 54 eyes. AB - OBJECTIVE: To document the 1-year postoperative clinical results of the safety, efficacy, and stability of conductive keratoplasty (CK) to correct low to moderate hyperopia. DESIGN: Prospective, nonrandomized, self-controlled trial. PARTICIPANTS: Five surgeons at four centers performed CK on the first 54 eyes of a multicenter, 2-year clinical trial. Treated eyes had +0.875 to +4.00 diopters (D) of hyperopia and < or =0.75 D of cylinder. The nature of this procedure was explained to all participating patients who signed informed consent forms prior to undergoing the procedure. INTERVENTION: Low energy, high-frequency current was applied directly into the peripheral corneal stroma through a delivery tip inserted at 16 or more treatment spots. An early nomogram was used for the first 54 eyes with an intended refraction of plano. MAIN OUTCOME MEASURES: Data from all 54 eyes were analyzed for safety and stability. A subset of 30 of the 54 eyes was found that had been treated with the appropriate number of spots with the early nomogram. These eyes were categorized as current nomogram eyes; the data for these eyes were analyzed for efficacy and predictability, as well as stability and safety. All patients reported on satisfaction and quality of vision after surgery. RESULTS: Preoperatively, the manifest refractive spherical equivalent refraction (MRSE) of the 30 current nomogram eyes was +1.57 D. At 1 year postoperatively, uncorrected visual acuity was 20/20 or better in 57% of the eyes and 20/40 or better in 93%. The MRSE was within 0.50 D in 46%, within +/ 1.00 D in 93%, and within +/-2.00 D in 100%. No eye lost > or =2 lines of best spectacle-corrected visual acuity at 1 year postoperatively nor had an induced cylinder of > or =2.00 D. The MRSE changed a mean of 0.25 D +/- 0.43 between 3 and 6 postoperative months, 0.16 D +/- 0.38 between 6 and 9 postoperative months, and 0.07 D +/- 0.38 between 9 and 12 postoperative months. Refractive stability appeared to be attained by 6 months postoperatively. CONCLUSIONS: Conductive keratoplasty appears to be safe, effective, and stable for correcting low to moderate spherical hyperopia. Stability appeared by the 6-month follow-up visit. For the eyes treated with the current CK nomogram, uncorrected visual acuity, predictability, and stability are as good as or better than those obtained with hyperopic laser in situ keratomileusis or noncontact laser thermal keratoplasty. PMID- 11927420 TI - A comparative analysis of five methods of determining corneal refractive power in eyes that have undergone myopic laser in situ keratomileusis. AB - OBJECTIVE: To evaluate the accuracy of computerized videokeratography, keratometry, and the Gaussian optics formula for measuring corneal refractive power in patients after myopic laser in situ keratomileusis (LASIK). DESIGN: Noncomparative case series. PARTICIPANTS: One hundred eyes of 63 patients (mean age, 45.0 +/- 10.9 [standard deviation] years) who underwent LASIK were included in the study. METHODS: Using the clinical history method as the standard, we evaluated the accuracy of values of corneal refractive power derived from computerized videokeratography (the EffRP value of the EyeSys Corneal Analysis System, which averages corneal refractive power over the central 3 mm), keratometry (K), the Gaussian optics formula (GauRP), and values of EffRP and keratometry as modified according to the amount of LASIK-induced refractive change. MAIN OUTCOME MEASURES: Correlation of measured corneal power values to those obtained using clinical history method (HisRP). RESULTS: Although the values for HisRP were significantly correlated with postoperative EffRP and K values and with GauRP, postoperative EffRP and K values were higher than HisRP (0.87 +/- 0.68 diopters [D] and 1.16 +/- 1.10 D, respectively), and GauRP were lower than HisRP (0.44 +/- 0.66 D) (P < 0.001 for all three comparisons). The differences between HisRP and both postoperative EffRP and K values increased significantly with the amount of myopic correction. The most accurate results were obtained by modifying the postoperative values of EffRP according to the amount of LASIK-induced refractive change; 70% of these values were within +/- 0.5 D and 94% within +/- 1 D of HisRP values. CONCLUSIONS: Using the clinical history method as the standard, we found that the most accurate method for determining corneal refractive power in post-LASIK eyes was to adjust the postoperative corneal measurement according to the amount of LASIK-induced refractive change. PMID- 11927421 TI - Steroid-induced glaucoma after laser in situ keratomileusis associated with interface fluid. AB - PURPOSE: To report the ocular manifestations and clinical course of eyes developing interface fluid after laser in situ keratomileusis (LASIK) surgery from a steroid-induced rise in intraocular pressure. DESIGN: Retrospective, noncomparative interventional case series. PARTICIPANTS/INTERVENTION: We examined six eyes of four patients who had diffuse lamellar keratitis develop after uneventful myopic LASIK surgery and were treated with topical corticosteroids. PRINCIPAL OUTCOME MEASURE: Slit-lamp findings, intraocular pressure measurements, and visual field loss. RESULTS: All eyes had a pocket of fluid develop in the lamellar interface between the flap and the stromal bed associated with a corticosteroid-induced rise in intraocular pressure. However, because of the interface fluid, intraocular pressure was normal or low by central corneal Goldmann applanation tonometry in all eyes. The elevated intraocular pressure was diagnosed by peripheral measurement in several cases after months of elevated pressure. All six eyes had visual field defects develop. Three eyes of two patients had severe glaucomatous optic neuropathy and decreased visual acuity develop as a result of undiagnosed steroid-induced elevated intraocular pressure. CONCLUSIONS: A steroid-induced rise in intraocular pressure after LASIK can cause transudation of aqueous fluid across the endothelium that collects in the flap interface. The interface fluid leads to inaccurately low central applanation tonometry measurements that obscure the diagnosis of steroid-induced glaucoma. Serious visual loss may result. PMID- 11927422 TI - Clear lens extraction with intraocular lens implantation during retinal detachment repair in patients with Acquired Immune Deficiency Syndrome (AIDS) [correction of autoimmune deficiency syndrome] and cytomegalovirus retinitis. AB - OBJECTIVE: To assess the outcomes of clear lens extraction with intraocular lens (IOL) implantation during repair of retinal detachment by vitrectomy with silicone oil tamponade in patients with acquired immunodeficiency syndrome (AIDS) and cytomegalovirus (CMV) retinitis. DESIGN: Retrospective, noncomparative case series. PARTICIPANTS: Twelve eyes of 10 patients with AIDS, CMV retinitis, and retinal detachment. INTERVENTION: All patients underwent phacoemulsification with posterior chamber IOL placement at the time of vitrectomy with silicone oil tamponade for repair of retinal detachment. A targeted postoperative refractive error of -5.00 diopters (D) to -3.00 D was chosen in an attempt to counteract the hyperopic effect of silicone oil. MAIN OUTCOME MEASURES: The following factors were evaluated: postoperative visual acuity, refractive error, and intraoperative and postoperative complications. RESULTS: Median follow-up was 7 months (range, 1 46 months). For patients without macular necrosis, median best-corrected preoperative visual acuity was 20/75 (range, 20/20-20/800), and median best postoperative visual acuity was 20/50 (range, 20/20-20/400). Median final visual acuity was 20/140 (range, 20/25 to count fingers at 1 foot). The median postoperative refractive error (spherical equivalent) was -1.00 D (range, -4.00 D to +7.88 D). Reoperation was required in 3 of 12 eyes for recurrent macular detachment (1 with silicone oil underfill; 2 with proliferative vitreoretinopathy). The macula was attached in all eyes at last follow-up. Reattachment of the peripheral retina was achieved in 10 of 12 eyes. There were no anterior segment complications. CONCLUSIONS: Clear lens extraction with IOL placement during repair of retinal detachment with silicone oil tamponade does not seem to increase complications and may improve long-term visual rehabilitation, improve retinitis management by allowing better posterior segment visualization throughout the postoperative course, and decrease overall cost and morbidity associated with cataract extraction as a second procedure. PMID- 11927424 TI - Multifocal intraocular lens implantation in prepresbyopic patients with unilateral cataract. AB - OBJECTIVE: To evaluate the benefits of implantation of a zonal-progressive multifocal intraocular lens (IOL) in prepresbyopic patients with unilateral cataract. STUDY DESIGN: Prospective, nonrandomized, comparative trial. PARTICIPANTS: Ninety-five eyes of 95 prepresbyopic patients aged between 14 and 40 years with either multifocal or monofocal IOL implantation at two institutions and with more than 6 months follow-up. INTERVENTIONS: Temporal clear corneal phacoemulsification and foldable IOL implantation was performed in all eyes. In 54 patients, a zonal-progressive optic multifocal IOL (Array SA40-N, Allergan, Irvine, CA) was implanted, whereas 41 patients received a monofocal IOL and served as controls. RESULTS: Preoperative patient demographics, mean postoperative spherical equivalent, astigmatism, and uncorrected and best corrected distance visual acuities were similar in the two groups. Patients with a multifocal IOL achieved a significantly better uncorrected near visual acuity than patients with monofocal IOL (0.18 versus 0.37; P = 0.0001). With distance correction only, mean near visual acuity was 0.17 versus 0.43 (P = 0.0001). Best corrected near visual acuity was 0.11 for both groups (P = 0.91), with +1.43 diopters (D) for the multifocal group and +2.35 D for the monofocal group (P = 0.0001). Spectacle dependency differed significantly between the two groups, with 21 patients (51%) of the monofocal group commonly requiring an additional plus add for near tasks compared with 5 patients (9%) in the multifocal group (P = 0.001). Stereopsis was superior in the multifocal group (P < 0.001), with 42 (77%) and 33 (61%) patients with a multifocal IOL responding positively to the Lang and Titmus tests, respectively. In the monofocal group; only 20 (48%) and 9 (22%) patients gave correct answers. CONCLUSIONS: Multifocal IOL implantation is a viable alternative to monofocal pseudophakia in prepresbyopic patients with unilateral cataract. PMID- 11927423 TI - Phototherapeutic keratectomy versus diamond burr polishing of Bowman's membrane in the treatment of recurrent corneal erosions associated with anterior basement membrane dystrophy. AB - PURPOSE: To compare the efficacy of phototherapeutic keratectomy (PTK) to epithelial debridement and polishing of Bowman's membrane using a diamond burr (DB) in the treatment of recurrent corneal erosions associated with anterior basement membrane (ABM) dystrophy. DESIGN: Retrospective nonrandomized comparative trial. METHODS: Medical records of 39 patients (42 eyes) who underwent a PTK or DB procedure for recurrent corneal erosions associated with ABM dystrophy between March 1992 and June 2000 were reviewed. History of injury, prior treatment received, and the corneal slit-lamp findings were noted in all patients. In both procedures, all loose epithelium was completely removed. In PTK patients, 5 microm of Bowman's membrane was ablated with the excimer laser. In patients who underwent DB treatment, a hand-held battery-driven diamond dusted burr was used to gently and uniformly polish Bowman's membrane in the area of the epithelial defect. MAIN OUTCOME MEASURES: Symptomatic improvement, recurrence of painful erosions, development of haze, and change in the visual acuity. Statistical analysis was performed for comparison of data between the two groups. RESULTS: Fifteen eyes of 14 patients underwent PTK, and 27 eyes of 25 patients underwent epithelial debridement and DB treatment. In the PTK group, mild haze was seen in five (35.7%) eyes. Recurrence of painful erosions was seen in four eyes (26.7%). The mean follow-up was 17.6 +/- 5.7 months (range, 0.7-81.9 months). Best-corrected visual acuity was better in five eyes (35.7%) after the procedure and the same in nine eyes (64.3%). In the DB group, mild haze was seen in seven eyes (25.9%). Recurrence of painful erosions was seen in three eyes (11.1%). The mean duration of follow-up was 6.7 months (range, 1-24.2 months). Best-corrected visual acuity was better in 3 eyes (14.3%), the same in 17 eyes (81%), and worse in 1 eye (4.8%) after the procedure. Final visual acuity was not available for one eye in the PTK group and 6 eyes in the DB group. There was no statistically significant difference in haze (Fisher's exact test, P = 0.38), recurrence of erosions (Kaplan-Meier analysis with log rank, P = 0.73), and vision being better or the same (Fisher's exact test, P = 0.6) between the PTK and DB groups. CONCLUSIONS: Both PTK and DB treatment are effective methods of treating recurrent corneal erosions associated with ABM dystrophy. Diamond burr treatment, being a simpler, less expensive office procedure with a tendency toward lesser incidence of haze and recurrence in this study, seems to have advantages over PTK in the treatment of recurrent corneal erosions. Further prospective studies are required to confirm the long-term efficacy of DB treatment in the management of recurrent corneal erosions associated with ABM dystrophy. PMID- 11927425 TI - Rapid direct antibiotic susceptibility testing in endophthalmitis. AB - OBJECTIVE: To compare a new rapid antibiotic susceptibility test (RAST) to the conventional method in patients with endophthalmitis. DESIGN: Prospective nonrandomized comparative trial. PARTICIPANTS: Intraocular aspirates from 24 consecutive patients with endophthalmitis were tested. METHODS: Approximately 0.25 ml of vitreous or anterior chamber aspirate was obtained and tested for antibiotic sensitivity using the Kirby-Bauer agar disk diffusion method. Using this conventional testing method, the aspirates were cultured for bacterial growth before antibiotic susceptibility testing (AST). In contrast, for direct RAST, the aspirates were inoculated directly onto agar plates with antibiotic disks without first culturing for bacterial growth. RESULTS: Of the 24 endophthalmitis aspirates studied, 4 (17%) could not be tested by direct RAST because either more than one bacterial species was present or no bacteria were cultured. The remaining 20 cases provided 467 antibiotic-microorganism combinations. The two methods yielded identical susceptibility results in 409 (88%) of the 467 tests. In 42 tests (9%), a minor discrepancy occurred between the conventional method and RAST, in which one resulted in intermediate susceptibility, whereas the other was either resistant or sensitive. Significant discrepancy, defined as a bacterial sample reported as sensitive in one method yet resistant in the other, occurred with 16 samples (3%). Results of the RAST were available within 6 to 10 hours compared with more than 24 hours for the conventional method. Of clinical significance, the results of the RAST revealed that in 11 cases of epidemic Stenotrophomonas maltophilia endophthalmitis, the bacteria were resistant to the antibiotics prescribed, and change of antibiotic therapy was made on the basis of the RAST results. CONCLUSIONS: RAST provided accurate antibiotic susceptibility results in a much shorter time than did the conventional method. We suggest that conventional AST be performed for confirmation of the RAST results, but initial antibiotic therapy can quite reliably be based on the results of RAST. PMID- 11927426 TI - Amniotic membrane grafts for nontraumatic corneal perforations, descemetoceles, and deep ulcers. AB - PURPOSE: To describe the clinical outcome of amniotic membrane transplantation (AMT) for nontraumatic corneal perforations, descemetoceles, and deep ulcers. DESIGN: Retrospective, noncomparative, interventional case series. PARTICIPANTS: Thirty-four eyes of 33 consecutive patients operated on for nontraumatic corneal perforations or descemetoceles at four academic departments of ophthalmology. Associated autoimmune disorders included rheumatoid arthritis (n = 6), Stevens Johnson syndrome (n = 3), ocular cicatricial pemphigoid (n = 2), systemic lupus erythematosus (n = 1), and one eye with Mooren's ulcer, as well as neurotrophic, or exposure keratopathy (n = 10), postinfectious nonhealing ulcers (n = 6), and postsurgery (n = 5). INTERVENTION: Three or four layers of amniotic membrane (AM) were applied over the ulcer bed and anchored with 10-0 nylon interrupted or running sutures. A large AM piece was used as a patch to cover the entire corneal surface. MAIN OUTCOME MEASURES: Formation of anterior chamber depth, epithelialization of the AM grafts, and stability of the corneal stromal thickness. RESULTS: The mean follow-up period was 8.1 +/- 5.7 (ranging from 2-23) months. A successful result was observed in 28 of 34 eyes (82.3%). Of the successful cases, 23 eyes needed one AMT procedure, whereas 5 eyes needed two procedures to achieve a successful result. In five eyes, a subsequent definitive surgical procedure such as penetrating keratoplasty or lid surgery was needed. Failure was observed in six eyes with rheumatoid arthritis, neurotrophic keratopathy, or graft melting. CONCLUSIONS: AMT is an effective method for managing nontraumatic corneal perforations and descemetoceles. It can serve as either a permanent therapy or as a temporizing measure until the inflammation has subsided and a definitive reconstructive procedure can be performed. This treatment option is also beneficial in those countries where corneal tissue availability is limited. PMID- 11927427 TI - Prevalence and progression of myopic retinopathy in an older population. AB - OBJECTIVE: To determine the prevalence and progression of myopic retinopathy in an older community-based population sample. DESIGN: Population-based epidemiologic study. PARTICIPANTS: Eligible residents aged 49 years or older (n = 3654) who attended the Blue Mountains Eye Study, west of Sydney, Australia. METHODS: Participants had a detailed eye examination including measurement of logarithm of the minimum angle of resolution (logMAR) visual acuity, standardized refraction, and retinal stereophotography. All patients were invited to attend follow-up examinations after 5 years. MAIN OUTCOME MEASURES: Myopic retinopathy was defined to include staphyloma, lacquer cracks, Fuchs' spot and myopic chorioretinal atrophy. beta-peripapillary atrophy was assessed separately. RESULTS: Signs of myopic retinopathy were found in 67 eyes from 44 participants (1.2%), a prevalence of 1.4% in women and 1.0% in men; this increased from 1% in right eyes with myopia <3 diopters to over 50% in right eyes with myopia > or =9 diopters. There was a nonsignificant age-related trend in prevalence. The average spherical equivalent refraction was -6.1 diopters and the average visual acuity was 20/40 in eyes with myopic retinopathy. Visual impairment (<20/40) was present in 38.8% of affected eyes. Myopic retinopathy was bilateral in 52% of cases. Staphyloma was present in 26 participants (0.7%), bilateral in 35%, with a strong concordance of staphyloma location. Lacquer cracks were seen in 8 participants (0.2%), Fuchs' spot in 3 (0.1%), and chorioretinal atrophy in 7 (0.2%). Forty-six eyes (68.7%) with myopic retinopathy were reexamined after 5 years; 8.7% had new or increased numbers of lacquer cracks and 15.2% had new or expanded areas of chorioretinal atrophy. In those eyes developing lacquer cracks or chorioretinal atrophy, best-corrected visual acuity decreased by a mean of two LogMAR lines. CONCLUSIONS: This study documented the age and sex-specific prevalence of myopic retinopathy and 5-year progression in an older white population. PMID- 11927428 TI - Long-term visual prognosis of choroidal neovascularization in high myopia: a comparison between age groups. AB - OBJECTIVE: To analyze visual outcome in highly myopic patients of different age groups with choroidal neovascularization (CNV). DESIGN: Retrospective observational case series. PARTICIPANTS: We reviewed the medical records of 63 consecutive patients (73 eyes) with myopic CNV. The patient population was divided into two groups according to age at onset of CNV (< or =40 and >40 years old). INTERVENTION: Demographic and clinical data were obtained from the patients' medical records. MAIN OUTCOME MEASURES: Visual acuity at least 3 years after CNV diagnosis. RESULTS: Group 1 (< or =40 years old) consisted of 22 patients (26 eyes), and group 2 (>40 years old) consisted of 41 patients (47 eyes). Throughout the follow-up period, group 1 retained better visual acuity than group 2. Almost half the patients in group 1 retained a final visual acuity better than 20/40. No significant change occurred in the logarithm of the minimum angle of resolution (logMAR) in group 1 during follow-up. Group 2 had worse visual acuity at the initial evaluation than did group 1, and a statistically significant worsening of logMAR was found during the follow-up period. More than half of the patients in group 2 had a final visual acuity less than 20/200. In addition, group 2 had a larger area of CNV, and chorioretinal atrophy was more frequently seen after the regression of CNV than in group 1. CONCLUSIONS: The visual prognosis of myopic CNV is influenced by age at onset. The results of this study indicate that patient age at the time of onset of myopic CNV should be considered when determining the therapeutic course. PMID- 11927429 TI - Assessment of vision-related function in patients with age-related macular degeneration. AB - OBJECTIVE: To investigate the validity of the visual function index (VF-14) in assessing visual function in patients with age-related macular degeneration (AMD). DESIGN: Prospective noncomparative observational case series. PARTICIPANTS: One hundred fifty-nine consecutive patients attending a sole practitioner's academic retina-only clinic from May 1998 through August 1998 and from May 1999 through August 1999. MAIN OUTCOME MEASURES: Correlations were calculated between the VF-14 scores and the medical outcomes study 36-item short form (SF-36), weighted comorbidity scale, visual acuity and clinical AMD severity (stage), and vision self-assessment scales. Documentation of the severity of macular degeneration was performed by a sole examiner. RESULTS: There was a moderately strong correlation between visual acuity and trouble with vision (r = 0.51), satisfaction with vision (r = -0.50), and overall quality of vision (r = 0.56). A strong correlation was noted between VF-14 score and patients' self rating of amount of trouble with vision (r = -0.67), satisfaction with vision (r = 0.62), and overall quality of vision (r = 0.67). In comparison, correlations between SF-36 score and patients' self-rating of amount of trouble with vision, satisfaction with vision, and overall quality of vision ranged from r = 0.37 to r = -0.40. Linear regression analysis for the overall study population indicated that AMD severity was not an independently significant predictor of VF-14 score after adjusting for visual acuity. However, among patients with 20/20 vision in the better eye, AMD severity was an independently significant predictor of VF-14 score after adjusting for visual acuity in the worse eye. CONCLUSIONS: The VF-14 exhibits a considerable degree of validity as a measure of functional impairment in patients with AMD. Age-related macular degeneration severity was an independently significant predictor of VF-14 score in the group of patients with 20/20 vision in the better eye, but this did not hold true for the overall study population. Age-related macular degeneration is associated with substantial impairment in reported visual function. PMID- 11927430 TI - Genetic influence on early age-related maculopathy: a twin study. AB - OBJECTIVE: Age-related macular degeneration (AMD) is the most common cause of blindness in industrialized countries. There has been considerable interest in the genetics of early age-related maculopathy (ARM) and AMD, because they have phenotypes similar to inherited diseases where mutations have been identified, but the heritability of ARM and AMD is unknown. DESIGN: A classical twin study was performed to compare the concordance in monozygotic (MZ) and dizygotic (DZ) twins in an unselected sample of female volunteer twins. PARTICIPANTS: Five hundred six twin pairs, 226 MZ and 280 DZ, with a mean age of 62 years, were examined. METHODS: ARM was graded from stereoscopic macular photographs of 501 of the twin pairs (99%) according to the International ARM Epidemiologic Study Group grading system. The casewise concordance was calculated for twin pairs from 2 x 2 contingency tables of affected/unaffected twins, and these tables were used in maximum likelihood genetic modeling to estimate the heritabilities of phenotypes graded. MAIN OUTCOME MEASURES: Prevalence of ARM; concordance in MZ and DZ twins of the phenotypes of ARM, soft drusen >63 microm and > or =125 microm diameter, pigmentary changes and hard drusen (<20 and > or =20 in number); heritability of ARM and subphenotypes. RESULTS: The overall prevalence of ARM was 14.6% (95% confidence interval [CI], 12.4%-16.8%). The concordance for ARM in MZ twins was 0.37 compared with 0.19 in DZ twins, suggesting a role for genes. Modeling confirmed a genetic effect for phenotypes of ARM, soft drusen, pigmentary changes, and > or =20 hard drusen, although there was little genetic effect for scattered (<20) hard drusen. The heritability of ARM was estimated as 45% (95% CI, 35%-53%). The most heritable phenotypes were soft drusen > or =125 microm (57%) and > or =20 hard drusen (81%), with the latter being dominantly inherited. CONCLUSIONS: This study confirms a significant genetic influence in ARM and suggests that future genetic studies should examine phenotypes of large (> or =125 microm) soft drusen and > or =20 hard drusen, because these seem to be the most heritable components. PMID- 11927431 TI - Causes of blindness and visual impairment in a population-based sample of U.S. Hispanics. AB - OBJECTIVE: To describe the causes of blindness and visual impairment in a population-based sample of Hispanics. DESIGN: A cross-sectional study. PARTICIPANTS: A random sample of 4774 Hispanic residents of Santa Cruz and Pima Counties in Southern Arizona aged 40 years and older who participated in Proyecto VER (Vision Evaluation and Research). TESTING: Subjects were interviewed and underwent a thorough ophthalmic examination. Presenting and best-corrected visual acuity was determined using the Early Treatment of Diabetic Retinopathy Study protocol, followed by a standardized ophthalmic examination to determine the causes of visual loss. Anterior and posterior segment specialists in ophthalmology confirmed the causes. MAIN OUTCOME MEASURES: Causes of visual loss (best-corrected acuity worse than 20/40). RESULTS: The response rate of eligible participants was more than 70%. Best-corrected acuity in the better seeing eye worse than 20/40 increased from 0.3% in those aged 40 to 49 to 5.6% in those aged 65 and older. The leading cause was cataract, accounting for 42% of all visual loss, followed by age-related macular degeneration (15%), and diabetic retinopathy (13%). Among 14 people who were bilaterally blind, open-angle glaucoma was the leading cause. Women had higher age-adjusted prevalence of severe cataract compared with men and were more likely to be visually impaired from cataract, diabetic retinopathy, and open-angle glaucoma, although gender differences were not statistically significant. CONCLUSIONS: Causes of visual impairment differ from those reported in Caucasian populations, with open-angle glaucoma being the leading cause of blindness. Further work on gender-based obstacles to eye care in the Hispanic community may be warranted. PMID- 11927432 TI - Embolic central retinal artery occlusion detected by orbital color Doppler imaging. AB - OBJECTIVE: To describe the detection of emboli to the retrobulbar circulation using orbital color Doppler imaging in patients with central retinal artery occlusions (CRAO) without visible retinal emboli. DESIGN: Noncomparative, retrospective, observational case series. PATIENTS AND METHODS: Twenty-nine patients with CRAO, without funduscopic evidence of embolic material within the retinal arterioles, underwent neuroophthalmic examination and evaluation with orbital color Doppler imaging. MAIN OUTCOME MEASURES: The detection of retrobulbar plaques using orbital color Doppler imaging. RESULTS: Nine of 29 (31%) patients with CRAO demonstrated hyperechoic retrobulbar plaques. All nine patients had visual acuity of counting fingers or worse at presentation. Six of the nine patients (67%) had a history of hypertension, and seven (78%) had coronary artery disease as vascular risk factors for arterial occlusive disease. Two of the nine patients (22%) subsequently died from cardiac-related events. CONCLUSIONS: This study demonstrates that orbital color Doppler imaging is an important diagnostic procedure for establishing embolism as the cause of CRAO when no emboli are visible in the retinal circulation. This noninvasive technology enables prompt differentiation of embolic disease from arterial occlusion caused by intrinsic atherosclerosis, vasospasm, or vasculitis from giant cell arteritis. Recognition of emboli has important management implications for these patients. PMID- 11927435 TI - Orbital rhabdomyosarcoma metastatic to the contralateral orbit: a case report. AB - OBJECTIVE: To report a rare presentation of metastatic orbital rhabdomyosarcoma and the corresponding findings on magnetic resonance imaging (MRI). DESIGN: Interventional case report. RESULTS: A 29-year-old white man was diagnosed with rhabdomyosarcoma of the left sinus and orbit for which he was treated with chemotherapy and radiation. Eighteen months after diagnosis, he returned with subacute right eye pain and dysmotility of his extraocular muscles. MRI revealed solitary enlargement of the right medial rectus muscle, and thyroidopathy was suspected. Over the next 2 months, symptoms progressed, and proptosis developed. MRI showed infiltration of seven extraocular muscles. A biopsy of right orbital tissues and the right medial rectus muscle was performed. Special tissue typing confirmed metastatic alveolar rhabdomyosarcoma. The patient underwent palliative radiation therapy and chemotherapy, but he ultimately died of disseminated disease. CONCLUSIONS: Rhabdomyosarcoma can rarely metastasize to the extraocular muscles. Earlier recognition of orbital metastasis through radiographic and biopsy findings, along with prompt and aggressive treatment, may prevent fulminant spread of rhabdomyosarcoma. PMID- 11927434 TI - Chlamydia pneumoniae seropositivity and the risk of nonarteritic ischemic optic neuropathy. AB - OBJECTIVE: To determine whether IgG antibodies to Chlamydia pneumoniae are associated with nonarteritic ischemic optic neuropathy (NAION). DESIGN: Retrospective case-control study. PARTICIPANTS: The study cohort consisted of 71 consecutive patients with NAION and 71 controls matched for age and gender. MAIN OUTCOME MEASURES: Serum immunoglobulin G (IgG) antibody titers to Chlamydia pneumoniae. RESULTS: Patients with NAION had significantly higher IgG antibody titers to C. pneumoniae compared with control subjects (IgG titer > or =1:128: 29 patients versus 15 controls, P = 0.017). The odds ratio for patients with an IgG titer > or =1:128 was 2.56 (95% confidence interval [CI], 1.2-5.5). Adjustment for arterial hypertension, diabetes mellitus, and myocardial infarction resulted in an odds ratio of 3.48 (95% CI, 1.3-9.6). CONCLUSIONS: Our results suggest that elevated titers of IgG antibodies to C. pneumoniae are associated with NAION. PMID- 11927436 TI - Learning effects among perimetric novices in frequency doubling technology perimetry. AB - PURPOSE: To investigate learning effects, including improvements in reliability indices, in frequency doubling technology (FDT) perimetry in subjects without glaucoma who have not undergone previous automated perimetry. DESIGN: Prospective consecutive case series. PARTICIPANTS: Eighty-one normal subjects. METHODS: Normal subjects were recruited from the University of Washington Eye Clinic who had no history of automated visual field testing. All eyes underwent complete ophthalmic examination. The C-20-5 screening mode of FDT perimetry (Zeiss Humphrey, San Leandro, CA) was performed after appropriate instruction, including a demonstration of the frequency doubling phenomenon. If any abnormal areas were identified or there were greater than 33% fixation losses or false-positive results, then the test was readministered up to three times or until a normal result was returned. MAIN OUTCOME MEASURES: Normal FDT test. RESULTS: The mean age was 54.8 years (range, 19-94). Sixty-nine subjects (85.2%) returned a normal test after a single test administration. Nine subjects (11.1%) required two administrations, and two subjects (2.5%) required three or more administrations to return a normal test. One subject still tested unreliably after four tests. The number or severity of abnormal locations did not predict the number of trials necessary to overcome learning effects. Fixation loss was the most common reliability problem. Subjects who demonstrated learning effects did not differ significantly in age, visual acuity, refraction, or test time from those who did not. CONCLUSIONS: Some perimetric novices demonstrate learning effects in C-20-5 screening-mode FDT perimetry. Learning effects in FDT perimetry must be considered during screening for glaucoma or other ocular disease. PMID- 11927437 TI - Predictive value of short-wavelength automated perimetry: a 3-year follow-up study. AB - PURPOSE: To determine whether structural changes in the retinal nerve fiber layer (RNFL) and functional abnormalities in short-wavelength automated perimetry (SWAP) can predict the onset of functional losses in standard automated perimetry patients suspected of having glaucoma. DESIGN: Prospective observational case series. PARTICIPANTS: A total of 160 eyes of ocular hypertensive subjects (intraocular pressure greater than 21 mmHg and normal standard automated perimetry) were included in the study. INTERVENTIONS: The subjects underwent RNFL photographic evaluation and SWAP. Standard automated perimetries were repeated after 3 years to evaluate glaucomatous losses. MAIN OUTCOME MEASURES: Onset of glaucomatous defects in conventional automated perimetry after 3 years of follow up, with or without prior glaucomatous defects in RNFL and SWAP. RESULTS: At the beginning of the study, 77 eyes showed RNFL losses (48%), and 58 eyes showed abnormalities in SWAP (36%). After the 3-year follow-up period, 14 of 77 eyes with RNFL losses had standard automated perimetry abnormalities (predicting sensitivity 93%), whereas 11 of 58 eyes with abnormal SWAP had standard automated perimetry losses (73% sensitivity). CONCLUSIONS: RNFL and SWAP losses are signs of early glaucomatous damage and can predict functional losses in standard automated perimetry. PMID- 11927438 TI - Long-term follow-up of visual field progression after trabeculectomy in progressive normal-tension glaucoma. AB - OBJECTIVE: To evaluate the long-term effects of trabeculectomy on the progression of visual field damage in patients with progressive normal-tension glaucoma (NTG). DESIGN: Retrospective, noncomparative, interventional case series. PARTICIPANTS: Twenty-three patients with NTG who had significant progression of visual field damage preoperatively and underwent trabeculectomy using antimetabolites. METHODS: Visual field testing using the Humphrey Visual Field Analyzer was periodically performed before and for at least 5 years after surgery (mean, 6 years). The time course of the mean deviation (MD) and mean of total deviations (TD(mean)) in four separate subfields, superior and inferior cecocentral and superior and inferior arcuate fields, were analyzed using a linear mixed effects model. MAIN OUTCOME MEASURES: Intraocular pressure (IOP), preoperative and postoperative regression coefficients of the time course of MD or TD(mean) in the four subfields, corresponding to the rate of progression of visual field damage. RESULTS: IOP significantly decreased from 16.2 +/- 1.8 mmHg preoperatively to approximately 11 mmHg during the postoperative follow-up period (P < 0.001). The preoperative regression coefficient of MD change was significantly negative (-1.05 [95% confidence interval, -1.28 to -0.82] dB/year, P < 0.001). After surgery, it increased significantly to -0.44 (95% confidence interval, -0.64 to -0.24) dB/year (P < 0.001), but the postoperative value was still significantly negative (P < 0.001). In the superior cecocentral, superior and inferior arcuate fields, preoperative regression coefficients of TD(mean) change were significantly negative (P < 0.001) and significantly increased after surgery (P < 0.01). In the inferior cecocentral field, the preoperative regression coefficient was not significantly different from 0 (P = 0.72) and did not change significantly after surgery (P = 0.15). CONCLUSIONS: Trabeculectomy was statistically associated with slowing further progression of visual field damage in patients with progressive NTG. The progression, however, did not completely stop over the 6-year postoperative follow-up period. PMID- 11927439 TI - Greater pupillary escape differentiates central from peripheral visual field loss. AB - OBJECTIVE: To test whether pupil escape observed during a constant light stimulus was greater in eyes with central visual field loss compared to eyes with peripheral visual field loss and normal eyes. DESIGN: Comparative, observational case series. PARTICIPANTS: Twenty-seven normal subjects, 5 patients with central field loss, 11 patients with peripheral field loss, and 8 patients with combined loss (central and peripheral visual field loss) were tested. METHODS: A dual channel infrared pupillograph was used to simultaneously record the right and left pupil diameters at a rate of 60 Hz to characterize the initial, phasic pupil contraction, and the sustained, or prolonged, pupil contraction in response to a 5-second light stimulus. Full-field light stimuli with a diameter of 30 degrees were presented to each eye at seven different intensities. MAIN OUTCOME MEASURES: The amplitude of the phasic pupil contraction was compared with the amplitude of the sustained pupil contraction at the four brightest intensities (slope of phasic versus sustained contraction) in the normal eyes and in eyes with visual field loss in the center and the periphery. RESULTS: The sustained pupillary contraction showed a statistically greater reduction in proportion to the phasic component in eyes with central field loss compared to those with peripheral field loss, combination central and peripheral field loss, and normal eyes. CONCLUSIONS: Afferent neurons from the central retina normally contribute a substantial component to the sustained pupil response. Eyes with central field loss can be distinguished from eyes with peripheral field loss by a relatively greater loss of the sustained response, causing greater pupil escape. This study showed that pupil movements in response to a nonperimetric full-field light stimulus may provide some degree of mapping of retinotopic sensitivity. This information may be of use in interpreting the pupillary movements during the swinging flashlight test and may help predict the pattern of visual field loss. PMID- 11927441 TI - Effects of viscoelastic injection into Schlemm's canal in primate and human eyes: potential relevance to viscocanalostomy. AB - OBJECTIVE: To study possible initial mechanisms of alterations in aqueous outflow that may result from the injection of viscoelastic into Schlemm's canal (SC) during viscocanalostomy. DESIGN: Experimental animal (Macaca nemestrina) and human autopsy study. METHODS: Eyes were dissected into a limbal ring and hemisected or quartered. Uncannulated segments served as controls. In treated segments, SC was cannulated and viscoelastic injected. Segments were fixed and sectioned into continuous 500 to 1000 microm pieces, examined at the dissecting microscope, and photomicrographs were taken. Representative tissue was further prepared for scanning electron microscopy or sectioned at 1 microm. MAIN OUTCOME MEASURES: Tissue sections were examined to determine the extent of dilation and disruption of SC and related structures. SC dimensions were measured in segments from controls, cannulated regions, and regions of viscoelastic injection beyond the cannula insertion. In the hemisected segments, the circumferential extent of SC dilation was determined, and structures within the canal were described and counted. RESULTS: SC was dilated with increased anteroposterior length and height in cannulated and viscoelastic-injected segments in both primate and human eyes relative to untreated controls. The walls of SC were disrupted in both regions of cannulation and of viscoelastic injection, and the collector channels were widely dilated by viscoelastic. With decreasing effectiveness, the injected viscoelastic circumferentially dilated SC as far as 14 mm and 16 mm in primate and human hemisections, respectively. Structures that bridged between the walls of SC were often disrupted. CONCLUSIONS: Cannulation of SC and injection of viscoelastic beyond the cannula resulted in marked dilation of SC and associated collector channels. Lateral walls, inner wall endothelium, and bridging structures of SC were frequently disrupted by cannulation and sometimes by injected viscoelastic. These findings suggest that viscocanalostomy may acutely cause a direct communication between SC and the juxtacanalicular space, and so may initially enhance conventional aqueous outflow. Controlled clinical trials will be necessary to determine the long-term outcomes of this procedure. PMID- 11927440 TI - Predictors for success of needle revision of failing filtration blebs. AB - OBJECTIVE: To assess the factors that can predict the success of external needle revision of failing blebs. DESIGN: Retrospective, nonrandomized, comparative case series. PARTICIPANTS: Forty-nine eyes of 43 patients who underwent a needle revision of a filtration bleb with 5-fluourouracil (5-FU) by one physician (SVLB) between August 1993 and December 1994. INTERVENTION: Needle revision of a failing filtration bleb using 5-FU. MAIN OUTCOMES: The intraocular pressure, glaucoma medications, and any complications. RESULTS: The risk ratio of fornix-based trabeculectomies compared with limbus-based trabeculectomies was 3.781; P = 0.047. The risk ratios for gender, race, type of glaucoma, lens status, previous surgery, antimetabolite used for the initial trabeculectomy, and preoperative bleb characteristics were not statistically different for any one group. CONCLUSIONS: Fornix-based trabeculectomies were more likely to fail the needle revisions compared with limbus-based trabeculectomies. None of the five preneedling bleb characteristics showed a propensity toward success or failure. Other demographics such as gender, race, type of glaucoma, previous surgery, antimetabolite used for the initial trabeculectomy, and lens status were also not predictive for success. PMID- 11927442 TI - Clinical outcome of eight BIGH3-linked corneal dystrophies. AB - OBJECTIVE: To determine whether the mutational pattern of BIGH3-linked corneal dystrophies (CDs) can accurately predict the clinical course of the disease and be helpful in planning adequate surgical treatment. DESIGN: Retrospective noncomparative case series. PARTICIPANTS: Chart review of 73 patients (110 eyes) with recently confirmed BIGH3 mutations who underwent a penetrating keratoplasty (PK) from 1978 through 1999. Diagnoses included Thiel-Benhke CD (TBCD/R555Q) (13 eyes), classic granular CD (CGCD/R555W) (28 eyes), superficial variant of granular dystrophy (SVGD/R124 l) (27 eyes), lattice CD type I (LCDI/R124C) (20 eyes), Avellino CD (ACD/R124H) (2 eyes), H626R-lattice dystrophy (LCD/H626R) (6 eyes), and two novel dystrophies: a French variant of granular dystrophy (FVGD/R124 l+DT125-DE126) (9 eyes) and a French lattice CD type IIIA (LCDIIIA/A546T) (5 eyes). METHODS: The mutation of the BIGH3 gene was characterized for all patients. Clinical data were reviewed for each patient, and included age at first PK and elapsed time before significant recurrence (as defined by a severe decrease in best-corrected visual acuity related to recurrent deposits in the graft). MAIN OUTCOME MEASURES: Mean age at first PK and delay before a significant recurrence. RESULTS: Mutational pattern was highly correlated with the clinical course of each dystrophy. According to the genetic mutation, two groups with different prognosis were identified. Group 1 was defined by the presence of the FVGD/R124 l+DT125-DE126 and SVGD/R124 l mutations and was characterized by the early need for treatment and early recurrence of deposits. Group 2 was molecularly defined by the presence of any of the following mutations: LCDI/R124C, CGCD/R555W, LCDIIIA/A546T, TBCD/R555Q, and LCD/H626R. In group 2, mean age at first treatment was older, and delay before a significant recurrence was longer as compared with group 1 (P = 0.0001). CONCLUSIONS: These results demonstrate that there is a direct correlation between the molecular defect and the clinical course of BIGH3-linked CDs. They also indicate that molecular characterization of the genetic defect will help predict and design adequate surgical treatment for patients with ambiguous clinical diagnosis. PMID- 11927443 TI - A prospective evaluation of subconjunctival injection of triamcinolone acetonide for resistant anterior scleritis. AB - PURPOSE: Prospective evaluation of the efficacy and safety of subconjunctival triamcinolone injections for resistant, nonnecrotizing, anterior scleritis. DESIGN: Prospective, noncomparative, interventional case series. PARTICIPANTS: Twelve eyes of 10 consecutive patients seen in the Doheny Eye Institute between August 1999 and December 2000 with diffuse, nonnecrotizing anterior scleritis that was resistant to systemic antiinflammatory therapy. Eyes with a history of steroid response were excluded. INTERVENTION: Subconjunctival injection of triamcinolone acetonide to the areas of maximal inflammation. MAIN OUTCOME MEASURES: Scleritis activity, adverse treatment effects, and number of systemic medications required at the end of the follow-up period. RESULTS: Within 1 to 14 days after injection, complete resolution of signs and symptoms was achieved in 11 eyes and partial resolution in 1 eye. Two patients had one recurrence each, 2.5 and 11 months after injection. Six of 10 patients were able to discontinue all systemic therapy (prednisone +/- immunosuppressive drugs); the remaining 4 needed continued oral therapy for systemic indications. Transient ocular hypertension and subconjunctival hemorrhage occurred in one eye each. Median follow-up period was 15 months. No eye developed necrotizing scleritis. CONCLUSIONS: Subconjunctival injection of triamcinolone acetonide is a safe and effective treatment for resistant, nonnecrotizing anterior scleritis. It provides rapid effect, is well tolerated, and may spare patients the significant complications and side effects of systemic corticosteroid and immunosuppressive therapy. PMID- 11927447 TI - A new optotype chart for detection of nonorganic visual loss. AB - OBJECTIVE: To develop a new optotype chart for detection of nonorganic decreased vision. DESIGN: Comparative observational case series. PARTICIPANTS: Optotype thresholds determination in three normal subjects and testing on 30 consecutive patients with unclear visual acuity loss. METHODS: A new optotype chart was developed, with an optotype minimum angle of resolution that is independent of size. In three normal subjects rank correlation was calculated between the optotype thresholds and the optotype sizes. A pocket chart was tested in a masked manner on 30 consecutive patients referred because of unclear visual acuity loss. MAIN OUTCOME MEASURES: Optotype thresholds and usefulness in clinical routine (positive and negative predictive values including 95% confidence intervals [CI]). RESULTS: The optotype thresholds did not correlate with the optotype sizes. Ten percent (3 of 30) of the patients referred because of unclear visual acuity loss had to be excluded, because their diagnosis remained unclear. All patients (16 of 16) with organic visual loss saw all optotypes sizes. Eighty-nine percent (10 of 11) of patients with nonorganic visual loss claimed to see only the larger optotypes. The positive predictive value for nonorganic visual loss of the new pocket chart was 100% (CI, 74%-100%); the negative predictive value was 94% (CI, 75%-100%). CONCLUSIONS: The new pocket chart seems to be useful for detection of visual loss caused by nonorganic disease. PMID- 11927446 TI - Oxybuprocaine induces a false-positive response in immunochromatographic SAS Adeno Test. AB - OBJECTIVE: To investigate whether a solution of oxybuprocaine hydrochloride, 0.4%, results in a false-positive response in an immunochromatographic SAS Adeno Test. DESIGN: Experimental study. CONTROLS: Physiologic saline and 2% lidocaine. TESTING: Each chemical (100 microl) was diluted in a transport medium. Five drops (200 microl) of the resultant solution were dispensed into the round sample well of a test device. Fifteen samples were tested in each group. MAIN OUTCOME MEASURES: Ten minutes after the start of the test, a colored line in the "specimen" portion of the test membrane was visually read as positive or negative by a masked technician. RESULTS: No positive reaction was observed in the control groups (physiologic saline and lidocaine). A false-positive reaction was observed in six samples (33.3%) in the oxybuprocaine group. The positive rate was significantly higher in the oxybuprocaine group compared with those in control groups (P = 0.0062, Fisher's extract probability test). CONCLUSIONS: Oxybuprocaine may induce a false-positive reaction in an immunochromatographic SAS Adeno Test. We recommend the use of lidocaine, instead of oxybuprocaine, for local anesthesia in taking eye swabs from patients with suspected adenovirus infection. PMID- 11927448 TI - Effects of delaying fluid resuscitation on an injury to the systemic arterial vasculature. AB - OBJECTIVES: To determine the effects of delaying fluid on the rate of hemorrhage and hemodynamic parameters in an injury involving the arterial system. METHODS: Twenty-one adult, anesthetized sheep underwent left anterior thoracotomy and transection of the left internal mammary artery. A chest tube was inserted into the thoracic cavity to provide a continuous measurement of blood loss. The animals were randomly assigned to one of three resuscitation protocols: 1) no fluid resuscitation (NR), 2) standard fluid resuscitation (SR) begun 15 minutes after injury, or 3) delayed fluid resuscitation (DR) begun 30 minutes after injury. All of the animals in the two resuscitation groups received 60 mL/kg of lactated Ringer's solution over 30 minutes. Blood loss and hemodynamic parameters were measured throughout the experiment. RESULTS: Total hemorrhage volume (mean +/- SD) at the end of the experiment was significantly lower (p = 0.006) in the NR group (1,499 +/- 311 mL) than in the SR group (3,435 +/- 721 mL) or the DR group (2,839 +/- 1549 mL). Rate of hemorrhage followed changes in mean arterial pressure in all groups. Hemorrhage spontaneously ceased significantly sooner (p = 0.007) in the NR group (21 +/- 14 minutes) and the DR group (20 +/- 15 minutes) than in the SR group (54 +/- 4 minutes). In the DR group, after initial cessation of hemorrhage, hemorrhage recurred in five of six animals (83%) with initiation of fluid resuscitation. Maximum oxygen (O2) delivery in each group after injury was as follows: 101 +/- 34 mL O2/kg/min at 45 minutes in the DR group, 51 +/- 20 mL O2/kg/min at 30 minutes in the SR group, and 35 +/- 8 mL O2/kg/min at 60 minutes in the NR group. CONCLUSIONS: Rates of hemorrhage from an arterial injury are related to changes in mean arterial pressure. In this animal model, early aggressive fluid resuscitation in penetrating thoracic trauma exacerbates total hemorrhage volume. Despite resumption of hemorrhage from the site of injury, delaying fluid resuscitation results in the best hemodynamic parameters. PMID- 11927449 TI - End-tidal carbon dioxide monitoring during procedural sedation. AB - OBJECTIVE: To prospectively determine whether end-tidal carbon dioxide (ETCO2) monitors can detect respiratory depression (RD) and the level of sedation in emergency department (ED) patients undergoing procedural sedation (PS). METHODS: This was a prospective observational study conducted in an urban county hospital of adult patients undergoing PS. Patients were monitored for vital signs, depth of sedation per the physician by the Observer's Assessment of Alertness/Sedation scale (OAA/S), pulse oximetry, and nasal-sample ETCO2 during PS. Respiratory depression was defined as an oxygen saturation <90%, an ETCO2 >50 mm Hg, or an absent ETCO2 waveform at any time during the procedure. The physician also determined whether protective airway reflexes were lost during the procedure and assisted ventilation was required, or whether there were any other complications. Rates of RD were compared with the physician assessment of airway loss and between agents using chi-square statistics. Spearman's rho analysis was used to determine whether there was a correlation between ETCO2 and the OAA/S score. RESULTS: Seventy-four patients were enrolled in the study. Forty (54.1%) received methohexital, 21 (28.4%) received propofol, ten (13.5%) received fentanyl and midazolam, and three (4.1%) received etomidate. Respiratory depression was seen in 33 (44.6%) patients, including 47.5% of patients receiving methohexital, 19% receiving propofol (p = 0.008), 80% receiving fentanyl and midazolam, and 66.6% receiving etomidate. No correlation between OAA/S and ETCO2 was detected. Eleven (14.9%) patients required assisted ventilation at some point during the procedure, all of whom met the criteria for RD. Pulse oximetry detected 11 of the 33 patients with RD. Post-hoc analysis revealed that all patients with RD had an ETCO2 >50 mm Hg, an absent waveform, or an absolute change from baseline in ETCO2 >10 mm Hg. CONCLUSIONS: There was no correlation between ETCO2 and the OAA/S score. Using the criteria of an ETCO2 >50 mm Hg, an absolute change >10 mm Hg, or an absent waveform may detect subclinical RD not detected by pulse oximetry alone. The ETCO2 may add to the safety of PS by quickly detecting hypoventilation during PS in the ED. PMID- 11927452 TI - Response time effectiveness: comparison of response time and survival in an urban emergency medical services system. AB - Emergency medical services (EMS) administrators seek methods to enhance system performance. One component scrutinized is the response time (RT) interval between call receipt and arrival on scene. While reducing RTs may improve survival, this remains speculative and unreported. OBJECTIVE: To determine the effect of current RTs on survival in an urban EMS system. METHODS: The study was conducted in a metropolitan county (population 620,000). The EMS system is a single-tier, paramedic service and provides all service requests. The 90% fractile RT specifications required for county compliance include 10:59 minutes for emergency life-threatening calls (priority I) and 12:59 minutes for emergency non-life threatening calls (priority II). All emergency responses resulting in a priority I or priority II transport to a Level 1 trauma center emergency department over a six-month period were evaluated to determine the relation between specified and arbitrarily assigned RTs and survival. RESULTS: Five thousand, four hundred twenty-four transports were reviewed. Of these, 71 patients did not survive (1.31%; 95% CI = 1.04% to 1.67%). No significant difference in median RTs between survivors (6.4 min) and nonsurvivors (6.8 min) was noted (p = 0.10). Further, there was no significant difference between observed and expected deaths (p = 0.14). However, mortality risk was 1.58% for patients whose RT exceeded 5 minutes, and 0.51% for those whose RT was under 5 minutes (p = 0.002). The mortality risk curve was generally flat over RT intervals exceeding 5 minutes. CONCLUSIONS: In this observational study, emergency calls where RTs were less than 5 minutes were associated with improved survival when compared with calls where RTs exceeded 5 minutes. While variables other than time may be associated with this improved survival, there is little evidence in these data to suggest that changing this system's response time specifications to times less than current, but greater than 5 minutes, would have any beneficial effect on survival. PMID- 11927450 TI - A randomized clinical trial of analgesia in children with acute abdominal pain. AB - OBJECTIVE: To evaluate the effects of intravenous morphine on pain reduction, physical examination, and diagnostic accuracy in children with acute abdominal pain. METHODS: A randomized, double-blind, placebo-controlled clinical trial was conducted at an emergency department of a tertiary care children's hospital. Children aged 5-18 years with abdominal pain of < or =5 days' duration, pain score > or =5 on a 0-10 visual analog scale, and need for surgical evaluation were eligible. Following the initial assessment, patients were randomized to receive either 0.1 mg/kg morphine or an equal volume of saline. The pediatric emergency medicine physician and surgical consultant independently recorded the areas of tenderness to palpation and percussion, and their diagnoses before the study medication and 15 to 30 minutes later. RESULTS: Sixty patients were enrolled, and 29 received morphine and 31 received saline. The demographic characteristics between the two groups were similar. The median reduction of pain score between the two study groups was 2 (95% CI = 1 to 4; p = 0.002). There was no significant change in the areas of tenderness in both study groups. Children with surgical conditions had persistent tenderness to palpation and/or percussion. There was no significant change in the diagnostic accuracy between the study groups and between the physician groups. All patients requiring laparotomy were identified and no significant complication was noted in the morphine group. CONCLUSIONS: Intravenous morphine provides significant pain reduction to children with acute abdominal pain without adversely affecting the examination, and morphine does not affect the ability to identify children with surgical conditions. PMID- 11927453 TI - Pediatric emergency department utilization within a statewide medicaid managed care system. AB - OBJECTIVES: To describe pediatric emergency department (ED) utilization within a Medicaid managed care system and to investigate the association between pediatric ED utilization and the type of medical home. METHODS: This was a retrospective cohort study, set at Rhode Island's statewide Medicaid managed care program (RIte Care). The study population was a random sample of 2,000 children aged 19 to 35 months at the conclusion of a period of continuous RIte Care enrollment from July 1, 1996, until June 30, 1997. For description of ED utilization, rates of utilization and percentages were calculated. Bivariate and multivariate analyses were used to determine the effect of type of medical home and individual/family characteristics on ED utilization. Logistic regression models were used to determine odds ratios for types of medical home while controlling for sociodemographic factors and practice level variation. RESULTS: Of the 1,988 children who were eligible for analysis, 791 children utilized the ED during the study year at a rate of 68 ED visits/100 person-years. Thirty-one percent of those visits were for injuries/poisonings. Logistic regression analysis demonstrated decreased ED utilization for patients enrolled in a staff-model health maintenance organization (HMO) (OR 0.34; 95% CI = 0.27 to 0.42) or a health center (OR 0.71; 95% CI = 0.53 to 0.95) compared with hospital-based clinics. There was no significant association between the receipt of preventive services and ED utilization. CONCLUSIONS: Despite the presence of a defined medical home, ED use was common among preschool children enrolled in the statewide Medicaid managed care program. The type of medical home was an important determinant of ED use in this population. PMID- 11927454 TI - Health care utilization of chronic inebriates. AB - Chronic inebriates often use emergency services, including the emergency department (ED), because they lack other resources or access to primary care. Because of their complicated medical needs, which are often exacerbated by acute intoxication and related illness or injury, a relatively small number of acutely intoxicated chronic inebriates can stretch ED resources and contribute to ED over crowding. OBJECTIVE: In order to address this, as well as overutilization of other county services, three county programs were developed (ethnic- and gender specific supportive housing; intensive street case management) to reduce emergency resource utilization while still providing a safe environment. This study determined the effectiveness of these programs. The authors hypothesized that program enrollment would reduce medical and detoxification (detox) expenditures for this patient population. METHODS: Pre- and postprogram comparisons were made on the number of detox and medical visits, insured days, and charges. Data were retrieved (with patients' written informed consent) from hospital and health plan billings and county databases. Descriptive statistics compared groups pre and post enrollment. Least-squares regression predicted total and non-inpatient medical charges. RESULTS: Complete data were available for 92 of 122 patients (mean age = 47 years; 60% Native American; 93% male); seven had severe illnesses or injuries, skewing the mean. However, there were significant overall reductions in the median numbers of yearly detox visits (10 to 1) and medical visits (11 to 8), and in median medical charges ($5,436 to $2,770) and total health care charges ($9,297 to $5,218). The median number of days insured increased (284 to 353). By regression analysis, injury was the most important preprogram predictor of medical charges; illness drives charges post-entry. Alcohol-related visits added to the model before entry but disappeared post entry. CONCLUSIONS: These programs reduced health care use for most patients. However, serious medical illness or injury in a small number of patients contributed heavily to resource utilization. PMID- 11927457 TI - Emergency physician interpretation of the electrocardiogram. PMID- 11927455 TI - Evaluating acceptance and understanding of risk in the emergency department: are all risk statements created equally? AB - OBJECTIVE: To determine whether mathematically equivalent but conceptually different presentations of risk from radioactive isotope exposure might affect the rate of agreement to participate in a hypothetical research study. METHODS: This was a prospective study of consenting English-speaking subjects more than 18 years of age who were asked whether they would agree to participate in a mock study when presented with six mathematically equivalent statements of research related risk. Participants were classified as recognizing the equivalence of the risk statements if they accepted all or refused all of the risk statements. RESULTS: Three hundred forty-six subjects were enrolled. There were 55 subjects (16%; 95% CI = 12% to 20%) who refused all of the six risk statements, and 23 participants (6.7%; 95% CI = 4% to 9%) who accepted all of the given risk statements. Most of the participants (77%; 95% CI = 73% to 82%) did not recognize that the six risk statements were equivalent and agreed to some, or did not understand some of the risk statements. In stepwise multivariate logistic regression, being white, being of older age, and having higher education were associated with a higher likelihood of accepting or rejecting all six of the risk statements. CONCLUSIONS: Ethnicity, age, and education affected whether patients recognized the equivalence of six risk statements for a hypothetical study of low dose radiation. Risk acceptance may be more likely when familiar concepts are used to express research risks. Researchers must focus on strategies that enhance the understanding of research risks and address the reasons for nonparticipation by subjects who are younger, of different ethnic or cultural background, or of lower education. PMID- 11927458 TI - Emergency medical services advanced life support response times: lots of heat, little light. PMID- 11927459 TI - Emergency medicine and critical care certification. PMID- 11927460 TI - "Profiles in patient safety": a new feature. PMID- 11927462 TI - Profiles in patient safety: sidedness error. AB - This case describes a 45-year-old woman with significant respiratory distress secondary to a left-sided pleural effusion that mandated an urgent thoracentesis. An adverse event occurred when the physician performed the procedure on the incorrect side of the patient. Results of the incident investigation followed by a discussion of medical errors models, common errors types, human factors considerations, and conditions that contribute to error are presented. Pertinent case-specific and general concepts of a system approach to reduce this type of medical error are discussed, and educational recommendations are offered. PMID- 11927463 TI - Advanced statistics: statistical methods for analyzing cluster and cluster randomized data. AB - Sometimes interventions in randomized clinical trials are not allocated to individual patients, but rather to patients in groups. This is called cluster allocation, or cluster randomization, and is particularly common in health services research. Similarly, in some types of observational studies, patients (or observations) are found in naturally occurring groups, such as neighborhoods. In either situation, observations within a cluster tend to be more alike than observations selected entirely at random. This violates the assumption of independence that is at the heart of common methods of statistical estimation and hypothesis testing. Failure to account for the dependence between individual observations and the cluster to which they belong can have profound implications on the design and analysis of such studies. Their p-values will be too small, confidence intervals too narrow, and sample size estimates too small, sometimes to a dramatic degree. This problem is similar to that caused by the more familiar "unit of analysis error" seen when observations are repeated on the same subjects, but are treated as independent. The purpose of this paper is to provide an introduction to the problem of clustered data in clinical research. It provides guidance and examples of methods for analyzing clustered data and calculating sample sizes when planning studies. The article concludes with some general comments on statistical software for cluster data and principles for planning, analyzing, and presenting such studies. PMID- 11927465 TI - Heliox as a temporizing measure for pediatric foreign body aspiration. AB - The authors report their experience with using heliox as a temporizing measure for an aspirated foreign body in a young child. PMID- 11927464 TI - The normal difference in bilateral indirect blood pressure recordings in hypertensive individuals. AB - OBJECTIVE: If there is a significant dichotomy between bilateral systolic blood pressure determinations in both arms in hypertensive individuals, it is often taken as a sign of underlying pathology. The primary objective was to determine what the normal variation might be for a significant difference between blood pressures in both arms. A clinically significant difference was chosen as a systolic blood pressure value greater than 10 mm Hg between the upper extremities, as is commonly quoted at the bedside and in the literature. METHODS: Bilateral indirect blood pressure determinations were obtained in 100 subjects with hypertension in the sitting position. The blood pressure was obtained by two observers, one the recorder and the other who obtained the blood pressure but was blinded to the actual values taken. The order of blood pressure determination in the first arm was determined prior to the study from a table of random numbers. The coefficient of variation in obtaining the blood pressure in each arm was determined in 5 of the 100 subjects. The age, sex, and handedness of each individual were recorded as demographic variables. RESULTS: The average left and right systolic blood pressures were 139 +/- 22 and 141 +/- 22 mm Hg, respectively. The average left and right diastolic blood pressures were 78 +/- 12 and 79 +/- 13 mm Hg, respectively. There was no significant difference between left minus right systolic or diastolic differences (paired t-test). There was no significant difference between systolic or diastolic blood pressures between gender or between left- and right-handed individuals (nonpaired t-test). The average coefficients of variation for taking right and left arm systolic pressures were 2.90% and 1.32%, respectively. Eighteen subjects (18%) had differences in systolic blood pressure between both arms exceeding 10 mm Hg (10% to 26%, 95% confidence interval). CONCLUSIONS: Differences of more than 10 mm Hg in indirect systolic blood pressure recordings between arms are frequent in asymptomatic hypertensive individuals and do not per se indicate any pathologic condition. In the right clinical situation, differences that are noted should be repeated and should be added to the total clinical picture when used to determine whether a pathologic condition is present. Assuming no significant aortic or subclavian disease in the population tested, the specificity of the bilateral blood pressure test was 82%. PMID- 11927467 TI - ECG competency-by whom, for whom? PMID- 11927468 TI - Report on the medical student view from ground zero--New York City, 2001. PMID- 11927469 TI - Observational studies and "real world" anesthesia pharmacoeconomics. PMID- 11927470 TI - Is there a future for xenon anesthesia? PMID- 11927471 TI - Desflurane improves the throughput of patients in the PACU. A cost-effectiveness comparison with isoflurane. AB - PURPOSE: In a pharmacoeconomic approach of anesthesia, postanesthesia care unit (PACU) occupancy can be chosen as a criteria of effectiveness to compare two anesthetic drugs with different rates of elimination and different costs of administration. Our objective was to develop a cost-effectiveness approach for the comparison of isoflurane (I) and desflurane (D). METHOD: In this prospective observational study, 68 patients aged 18-70 received either D or I for maintenance of anesthesia for inpatient abdominal procedures. Length of stay (LOS) in PACU was collected by a blinded observer. After the relationship between duration of surgery and LOS in PACU had been established in the 68 observed patients, we estimated the PACU occupancy according to duration of surgery and time of admission in PACU using a computer model of 204 consecutive patients, based on the hypothesis of an exclusive use of either D or I. Outcome measures were direct costs of the anesthesia procedure and occupancy of the PACU. RESULTS: The direct cost of the anesthetic was significantly higher with D than with I. This represents an increase of CAN$ 2 708 for the 204 patients. PACU occupancy was reduced by at least one patient (out of five beds) during 26.1% of the time with D (P <0.01). DISCUSSION: Improving the throughput of patients in PACU by using new halogenated anesthetic agents with faster rates of elimination may outweigh the incremental cost of this strategy. This becomes particularly meaningful in operating theatres experiencing frequent overcrowded periods. PMID- 11927472 TI - Routine handling of propofol prevents contamination as effectively as does strict adherence to the manufacturer's recommendations. AB - PURPOSE: Propofol is a potential vector of infection, because it contains no preservative. Thus, the manufacturer's specific recommendations for preparing injections or infusions go beyond the guidelines commonly used in our operating rooms for preparing other iv drugs. The purpose of the present study was to determine whether in the daily routine of an operating theatre a modified propofol handling technique can prevent contamination as effectively as do the manufacturer's handling recommendations. METHODS: A total of 160 consecutive neurosurgical patients were allocated to either Group I (manufacturer's handling recommendations: i.e., 1) disinfecting propofol vials and ampoules before filling syringes; 2) replacing empty syringes; 3) discarding all material at the end of surgery); or Group II (modified propofol handling protocol: i.e., 1) refilling empty syringes; 2) renewing only the infusion line to the patient). RESULTS: Total contamination rates were comparable in both groups (Group I: 14/160 (8.75%), Group II: 13/160 (8.13%) (chi2= 0.074; P=0.96). Frequency of contamination was not different between groups; either in sample 1 taken at the beginning of the procedure, (Group I: 5/80 (6.25%) vs Group II: 6/80 (7.5%); chi2=0.098; P=0.76) or in sample 2, taken at the end, (Group I: 9/80 (11.25%) vs Group II: 7/80 (8.75%); chi2=0.278; P=0.598). CONCLUSION: We conclude that in the daily routine of the operating theatre following a modified propofol handling protocol prevents contamination of propofol syringes as effectively as does adhering to the manufacturer's specific handling recommendations. However, neither of the tested guidelines completely prevented contamination. PMID- 11927473 TI - Rocuronium pharmacokinetic-pharmacodynamic relationship under stable propofol or isoflurane anesthesia. AB - PURPOSE: To compare the pharmacokinetics, pharmacodynamics and the concentration effect relationship of rocuronium in patients under stable propofol or isoflurane anesthesia. METHODS: Ten patients were randomized to receive fentanyl, propofol and nitrous oxide (60%) or fentanyl, thiopental, isoflurane (1.2% end-tidal concentration) and nitrous oxide (60%). To obtain good intubation conditions and maintain adequate muscle relaxation during surgery, patients received two bolus doses of rocuronium: 0.5 mg x kg(-1) (1.7 x ED95) at induction followed one hour later by 0.3 mg x kg(-1) (1 x ED95). Arterial blood samples were obtained over six hours after the second bolus dose. Plasma concentrations of rocuronium were measured using high pressure liquid chromatography. Muscle twitch tension was monitored by mechanomyography for the two doses. Pharmacokinetic and pharmacodynamic parameters were determined. RESULTS: No differences in rocuronium pharmacokinetic parameters were observed between both groups. After the second bolus, clinical duration was 20 +/- 6 min in the propofol group vs 39 +/- 8 min in the isoflurane group (P <0.05). The effect compartment concentration corresponding to 50% block, EC50, was higher under propofol anesthesia: 1008 vs 592 microg x L(-1) (P <0.05). CONCLUSION: Rocuronium body disposition is similar under stable propofol or isoflurane anesthesia. In contrast to isoflurane, propofol does not prolong the neuromuscular block. Therefore, the potentiating effect of isoflurane is of pharmacodynamic origin only, as explained by an increased sensitivity at the neuromuscular junction. In contrast with isoflurane anesthesia where the dose of rocuronium has to be decreased under stable conditions, no dose adjustment is required under propofol anesthesia. PMID- 11927474 TI - Midlatency auditory evoked potentials do not allow the prediction of recovery from general anesthesia with isoflurane. AB - PURPOSE: To investigate midlatency auditory evoked potentials (MLAEP) waveforms during recovery from anesthesia. The hypothesis was that MLAEP are sensitive variables to discriminate between states of consciousness and unconsciousness during emergence from anesthesia. METHODS: MLAEP were recorded in the awake state and during the wake-up phase from isoflurane anesthesia in 22 female patients undergoing ophthalmologic surgery. During emergence from anesthesia the changes in latency and amplitude of MLAEP components Na, Pa and Nb were compared with the awake level. The next day the patients were asked for explicit memory for the recovery period. RESULTS: In 72% of the patients the MLAEP waveforms were completely suppressed during isoflurane anesthesia. When the patients responded and opened their eyes spontaneously 38 +/- 12 min after anesthesia, the latencies of Na (18.3 +/- 1.2 vs 17.6 +/- 1.3; P = 0.013) and Nb (47.4 vs 7.1 vs 44.7 +/- 7.8; P = 0.048) remained prolonged compared with awake values. In contrast, the amplitudes NaPa and PaNb had regained baseline level. Nine patients had explicit memory for the immediate recovery period. However, there was no difference for any MLAEP component between patients with and without memory at any time. CONCLUSIONS: The persistent changes of MLAEP latency components Na and Nb indicated impaired auditory signal processing 38 min after isoflurane anesthesia. There was a marked intra- and inter-individual variability during reversal of the anesthetic induced MLAEP changes. This limits the prediction of recovery of consciousness in the individual patient during emergence from anesthesia. PMID- 11927476 TI - Xenon and nitrous oxide do not depress cardiac function in an isolated rat heart model. AB - PURPOSE: To examine the inotropic and chronotropic effects of xenon (Xe) and nitrous oxide (N2O) compared with nitrogen (N2) on isolated rat hearts. The differences between Xe and N2O were also compared. METHODS: The effects of Xe, N2O and N2 on coronary perfusion pressure (CPP), heart rate, left ventricular developed pressure (LVDP) and double product (DP) were examined in isolated rat hearts perfused at constant flow (10 mL x min(-1)). Following stabilization and baseline measurement with 95% O2 (plus 5% CO2), the heart was exposed to buffer equilibrated with one of three test gases; 50% N2 with 45% O2 (Group N2: n=9), 50% Xe with 45% O2 (Group Xe: n=9), or 50% N2O with 45% O2 (Group N2O: n=9) for 30 min. Measurements were performed in the last minute of exposure to the test gases. RESULTS: Gas exposure in all three groups decreased O2 delivery (-50%), CPP (-11%), LVDP (-30%) and DP (-44%) compared with baseline values (P <0.001). However, there were no differences among the groups. CONCLUSION: Our data suggest that cardiac contractility was decreased by the effects of reduced O2 delivery, but both Xe and N2O did not cause further cardiac depressant effects compared to N2 in this experimental model. PMID- 11927475 TI - End-stage renal failure reduces central clearance and prolongs the elimination half life of remifentanil. AB - PURPOSE: To evaluate the pharmacokinetics of remifentanil in 13 end-stage renal failure patients compared to matched control patients with normal renal function. METHODS: Remifentanil was infused for 20 min at a rate of 0.1 microg x kg(-1) x min(-1). Serial arterial blood samples (3 mL) were drawn at the start of infusion (zero), five, ten, 15, 20, 22.5, 25, 27.5, 30, 35, 40, 45, 50, 55 and 60 min. Blood samples were immediately preserved with citric acid and chilled on ice. High performance liquid chromatography-tandem mass spectrometry concentration assay was performed using GI 95779B internal standard. RESULTS: A two-compartment pharmacokinetic model provided an adequate fit for individual patient data. There was no difference in the mean +/- SD distribution half life (t1/2) between the renal failure group (1.65 +/- 0.7 min) and the control group (1.58 +/- 0.54 min). There was a significant difference in the central clearance (Cl(c)) and elimination half life (t1/2) ss) between the renal failure group (28 +/- 7 mL x kg(-1) x min(-1) and 18.86 +/- 2.06 min, respectively) and the control group (46.3 +/- 13.8 mL x kg(-1) x min(-1) and 16.35 +/- 2.99 min, respectively). Remifentanil blood concentrations were significantly higher in the renal failure group than in the control group. CONCLUSION: We have demonstrated a significant reduction in the Cl(c) and a prolongation of t1/2 ss of remifentanil in end-stage renal failure patients. While statistically significant, these variations in the pharmacokinetics of remifentanil were clinically modest and may be explained by a reduced volume of distribution in the period following hemodialysis. PMID- 11927477 TI - Intra-synovial, compared to intra-articular morphine provides better pain relief following knee arthroscopy menisectomy. AB - PURPOSE: To evaluate the analgesic effect of morphine 1 mg administered into the synovial tissue and the outer third of the meniscus after knee arthroscopy. METHODS: In a prospective, double-blind, randomized study, 60 patients who required elective knee arthroscopy were assigned to two groups: Group A consisted of 30 patients who received a direct injection of morphine 1 mg into either the synovial tissue or the outer third of the meniscus following menisectomy. Group B consisted of 30 patients who received a direct injection of NaCl 0.9% 1 mL into the synovial tissue or the outer third of the meniscus following menisectomy. At the end of the operation Group A received an intra-articular injection of NaCl 0.9% 1 mL and bupivacaine 0.5% 10 mL and Group B received an intra-articular injection of morphine 1 mg and bupivacaine 0.5% 10 mL. Analgesic effect was evaluated by pain intensity (visual analogue scale; VAS) and analgesic requirements (paracetamol) during the first one, three, six, 12, 24, and 48 hr postoperatively. RESULTS: There was no significant difference between the two groups within the first six hours and after 24 hr following the operation regarding VAS score and analgesic requirements. At 12 and 24 hr following the operation, the VAS score and the analgesic requirements were significantly higher in Group B compared to Group A (P <0.01 and P <0.01, respectively). No patient developed side effects. CONCLUSION: We conclude that direct morphine injection into the synovia or the outer third of the meniscus provided better pain relief than intra-articular morphine after knee arthroscopy. PMID- 11927478 TI - Intravenous tenoxicam reduces uterine cramps after Cesarean delivery. AB - PURPOSE: Postpartum uterine contraction pain is a common phenomenon after Cesarean delivery. We investigated the effectiveness of tenoxicam in reducing uterine contraction pain. METHODS: We enrolled 120 consecutive non-breastfeeding women who were scheduled for elective Cesarean delivery. After the administration of spinal anesthesia with bupivacaine and intrathecal morphine 0.15 mg injection, the patients were randomly divided into two groups. Group I received placebo (normal saline) iv injection, and Group II received tenoxicam 40 mg iv injection after clamping the umbilical cord. Verbal analogue scale of wound pain and uterine contraction pain were recorded at two, four, eight,16, and 24 hr after Cesarean delivery. RESULTS: There was no significant difference in wound pain scores between the two groups (all scores < or =3). However, the tenoxicam group had significant lower uterine contraction pain scores and required less supplemental meperidine medication than did the placebo group (8.5% vs 41.4%, P <0.05). The incidences of nausea or vomiting, pruritus, and bleeding were not significantly different between groups. CONCLUSION: Intravenous tenoxicam 40 mg significantly reduced the intensity of uterine cramps in patients undergoing Cesarean delivery without increasing side effects. PMID- 11927479 TI - Validation of fentanyl pharmacokinetics in patients undergoing coronary artery bypass grafting. AB - PURPOSE: The current emphasis on more rapid recovery and earlier tracheal extubation after cardiac surgery requires greater precision in administering opioids to reap their benefits while minimizing the duration of postoperative respiratory depression. Therefore, we aimed to define a pharmacokinetic model that accurately predicts fentanyl concentrations before, during, and after cardiopulmonary bypass (CPB) in patients undergoing coronary artery bypass grafting (CABG). METHODS: Parameters for two-compartment and three-compartment models were estimated by applying population pharmacokinetic modelling to fentanyl concentration vs time data measured in 29 patients undergoing elective, primary CABG. The ability of these models to predict fentanyl concentrations in a second series of ten patients undergoing CABG was then assessed. RESULTS: A simple, three-compartment model had excellent predictive ability, with a median prediction error (PE = ([Fentanyl]meas - [Fentanyl]pred)/[Fentanyl]pred x 100%) of -0.5%, and a median absolute PE (APE = /PE/) of 14.0%. In comparison to the two-compartment models, linear regression of measured:predicted concentration ratios indicated that the three-compartment model was free of systematic and time related changes in bias (P < 0.05). The parameters of this three-compartment model are: V1 15.0 l, V2 20.0 l, V3 86.1 l, Cl1 1.08 L x min(-1), Cl2 4.90 L x min(-1), and Cl3 2.60 L x min(-1). CONCLUSIONS: Our pharmacokinetic model provides a rational foundation for designing fentanyl dose regimens for patients undergoing CABG. When combined with previously published information regarding intraoperative fentanyl pharmacodynamics, dose regimens that reliably achieve and maintain desired fentanyl concentrations throughout the intraoperative period can be designed to achieve specific therapeutic goals. PMID- 11927480 TI - Utility of esophageal Doppler as a minimally invasive hemodynamic monitor: a review. AB - PURPOSE: The current bedside "gold standard" for cardiac output (CO) monitoring is thermodilution using a pulmonary artery catheter (PAC) but there is a number of risks associated with its use. The primary objective of this review was to evaluate the utility of esophageal Doppler (ED) as a minimally invasive monitor of CO. SOURCE: Medline literature search from 1966 to 2001 with citation review for studies comparing ED to PAC thermodilution for CO in perioperative and critically ill patients. PRINCIPAL FINDINGS: Twenty-five publications were identified comparing ED and PAC measurement of CO in a broad range of patients. There was a good overall correlation between CO determined by ED and thermodilution (n = 18 studies, median R = 0.89, range 0.52 to 0.98) and minimal bias (n = 13, median -0.01, range 1.38 to 2 L x min(-1)). The precision of ED was only fair overall as assessed by limits of agreement. The ED technique was found to be responsive in detecting changes in thermodilution CO and was reliable demonstrating both low intra- and inter-observer variation. ED was reportedly easy to insert after minimal training and was safe, with no significant complications identified. CONCLUSION: ED is a practical, reliable, and valid device for measuring CO in perioperative and critically ill patients. Further studies with larger numbers of patients are needed to determine if the limited precision observed is inherent to the technique, the diagnoses of patients studied, or the small sample sizes. PMID- 11927481 TI - Multidisciplinary management of a Jehovah's Witness patient for the removal of a renal cell carcinoma extending into the right atrium. AB - PURPOSE: To highlight the management of a Jehovah's witness surgical patient presenting for cardiopulmonary bypass (CPB) and deep hypothermic circulatory arrest. CLINICAL FEATURES: A 47-yr-old male, Jehovah's Witness, with renal cell carcinoma was admitted for left radical nephrectomy and excision of tumour thrombus extending into the junction of the inferior vena cava (IVC) and right atrium (RA). The preoperative goals were to maximize red blood cell mass, delineate the extent of tumour extension and develop a surgical plan incorporating blood conservation strategies to minimize blood loss. A midline abdominal incision was made to optimize removal of the non-caval portion of the tumour from the intra-abdominal region. CPB and deep hypothermic circulatory arrest were instituted to aid in removing the tumour from the IVC and RA. Intraoperative blood conservation strategies included the use of acute normovolemic hemodilution, antifibrinolytics, cell salvage, point-of-care monitoring of heparin and protamine blood concentrations, leukocyte-depleting filter, and meticulous surgical techniques. The patient was successfully weaned from CPB and was transported to the cardiothoracic intensive care unit without complication. The patient was discharged home one week after the operation with a hemoglobin of 10.2 g x dL(-1) and a hematocrit of 31.2%. CONCLUSION: Multiple blood conservation techniques were employed to manage this Jehovah's Witness patient through complex cardiac surgery, which was previously denied to him at other institutions. The successful outcome of this patient, while respecting the right to refuse allogeneic blood products, is a result of a multidisciplinary collaboration as well as the application of established blood conservation techniques. PMID- 11927482 TI - Perioperative management of a patient presenting with a spontaneously ruptured esophagus. AB - PURPOSE: To report a case of spontaneous rupture of the esophagus and its anesthetic management. CLINICAL FEATURES: A 52-yr-old male presented with a seven day history of chest pain, respiratory distress, and swelling in the neck following forceful vomiting. Examination revealed hypotension, decreased air entry in the right lower lung field with crepitations, epigastric tenderness with abdominal distension and guarding of both right and left hypochondria. A contrast esophagogram showed extravasation of contrast material from the lower third of the esophagus into the mediastinum without pleural cavity involvement. Reinforced primary closure of a 5-cm transmural tear in the right anterolateral wall of the esophagus 5 cm above the gastro-esophageal junction was performed along with right-sided chest drainage. The anesthetic drugs and technique in this case were selected to avoid any increase in intra-abdominal pressure to prevent further spillage of gastric contents into the mediastinum through the perforation. Invasive monitoring was used to assess early hemodynamic changes and to administer fluid therapy and vasoactive drugs. Due to prolonged surgery, lung congestion, large fluid shifts, a long surgical incision and abnormal arterial blood gases, the patient was ventilated mechanically in the intensive care unit. Subsequently he developed an esophageal leak, septic shock, and multiple organ failure and died. CONCLUSION: In a patient with a spontaneous rupture of esophagus, the anesthetic considerations include avoidance of further aggravation of the esophageal tear, and resuscitation from a morbid inflammatory condition. PMID- 11927483 TI - Aspiration prevented by the ProSeal laryngeal mask airway: a case report. AB - PURPOSE: To describe a case of intraoperative passive regurgitation where the ProSeal laryngeal mask airway (PLMA) successfully protected the airway from the respiratory tract. CLINICAL FEATURES: A 32-yr-old man was electively scheduled for change of dressings and application of plaster of Paris to both legs. A size 5 PLMA was inserted on the first attempt and the patient allowed to breathe spontaneously. Twenty-five minutes into the procedure brown fluid was noticed in the drainage tube of the mask. There was no change in respiratory pattern nor any evidence of coughing retching or vomiting. Twenty-five millilitres of fluid were suctioned out of the tube which tested positive for acid. The PLMA was left in place and the procedure continued uneventfully. After removal of the mask pH testing showed the dorsum of the mask to have a pH of 7 and the ventrum/bowl of the mask to be dry with a pH of 7. The patient had no respiratory symptoms in the recovery room and the postoperative course was uneventful. CONCLUSIONS: This case illustrates that passive regurgitation can occur unexpectedly intraoperatively and shows that the PLMA can protect the airway during such an event by allowing the regurgitated fluid to pass up the drainage tube without leaking into the glottis. PMID- 11927484 TI - Best evidence in anesthetic practice. Prevention: dopamine does not prevent death, acute renal failure, or need for dialysis. PMID- 11927485 TI - Intraoperative mobile magnetic resonance imaging for craniotomy lengthens the procedure but does not increase morbidity. AB - PURPOSE: To evaluate anesthetic aspects of care provided for craniotomy using mobile intraoperative magnetic resonance imaging (iMRI). METHODS: Anesthetic factors were studied using a retrospective case-control design. The primary outcome measures were the duration of the surgical intervention; the recovery score and body temperature on arrival; and length of stay in the post-anesthetic care unit. Secondary outcome measures were estimated blood loss, perioperative transfusion requirements, and fluids administered. RESULTS: Seventy-six patients undergoing craniotomy in the MRI theatre were compared with a case-matched control group of patients who underwent neurosurgical interventions in the conventional operating room during the same time period. The only outcome measure that differed between the two groups of patients was the duration of surgery: the mean duration of procedures for patients who underwent imaging was 407 +/- 143 min compared to 285 +/- 122 min in the conventional operating theatre (P < 0.000). Actual time spent imaging accounted for approximately 100 min (83%) of the increased duration. CONCLUSION: Our results do not support concerns that the iMRI suite is a "hostile" environment for the delivery of anesthesia for craniotomy. With the exception of an increased duration of the procedure, patients undergoing anesthesia with iMRI showed no differences from those operated in the conventional operating theatres. PMID- 11927486 TI - Isoflurane does not further impair microvascular vasomotion in a rat model of subarachnoid hemorrhage. AB - PURPOSE: Since isoflurane is known to attenuate endothelium-dependent dilation (EDD) in normal cerebral arterioles, we examined whether the anesthetic has a similar effect and further impairs EDD in vessels exposed to SAH. METHODS: Autologous blood was introduced in the subarachnoid space and the parietal lobe harvested. Control animals were sacrificed without introduction of blood. The response of microvessles to the endothelium-dependent dilator adenosine diphosphate (ADP) 10(-9)-10(-4) M, the endothelium-independent dilator nitroprusside 10(-9)-10(-4) M, and ET-1 10(-13)-10(-8) M was measured by videomicroscopy in the presence of 0-2 minimum alveolar concentration (MAC) of isoflurane. RESULTS: Isoflurane attenuated EDD to ADP in control vessels [66 +/- 5% (control) vs 27 +/- 11% (2 MAC) dilation to ADP 10(-4) M, P < 0.05]. Although SAH was associated with reduced dilation to ADP, exposure to isoflurane did not further impair dilation to ADP after SAH [26 +/- 3% (SAH) vs 21 +/- 5% (SAH/2 MAC) dilation to ADP 10(-4) M, P = NS]. Dilation to nitroprusside was not affected by isoflurane or SAH. Constriction to ET-1 was reduced by 2 MAC of isoflurane [21 +/- 1% (control) vs 13 +/- 5% (2 MAC) constriction to ET-1 10(-8) M, P < 0.05], but not by 1 MAC of isoflurane in control vessels. Constriction to ET-1 was greatly attenuated by 1 or 2 MAC of isoflurane after SAH [32 +/- 5% (SAH) vs 18 +/- 4% (SAH/2 MAC) constriction to ET-1 10(-8) M, P < 0.05]. CONCLUSION: In rats, isoflurane does not further impair EDD after SAH and modulates the constrictive response to ET-1. Such an effect of isoflurane would not predispose the SAH-exposed vessels to vasospasm. PMID- 11927487 TI - New media. PMID- 11927488 TI - Problem with Ohmeda Excel 210 SE anesthetic machine. PMID- 11927489 TI - Dorsal continuation of the radial artery. PMID- 11927490 TI - A normal capnogram despite esophageal intubation. PMID- 11927491 TI - The intubating laryngeal mask airway in severe ankylosing spondylitis. PMID- 11927492 TI - Variation in BRCA1 cancer risks by mutation position. AB - Previous studies have reported variation in BRCA1 breast and ovarian cancer risks with mutation position, suggesting that mutations toward the 3' end of the gene are associated with lower ovarian cancer risks. We evaluated the evidence for genotype-phenotype correlations in 356 families with protein-truncating BRCA1 mutations. In contrast to previous reports, the ovarian:breast cancer ratio associated with mutations in a central region of the gene (nucleotides 2401-4190) was significantly higher than for other mutations [odds ratio, 1.70 (P = 0.017) compared with nucleotides 1-2400; odds ratio, 1.89 (P = 0.02) compared with nucleotides 4191-end]. The risks of breast and ovarian cancer conferred by mutations in different regions of the gene were estimated separately by conditional maximum likelihood. According to the best fitting model, the breast cancer risk associated with mutations in the central region was found to be significantly lower than for other mutations (relative risk, 0.71; 95% confidence interval, 0.58-0.86; P = 0.0002), whereas the ovarian cancer risk associated with mutations 3' to nucleotide 4191 was significantly reduced relative to the rest of the gene (relative risk, 0.81; 95% confidence interval, 0.66-1.00; P = 0.044). The cancer risks associated with missense mutations in the RING domain in exon 5 appear to be similar to those associated with protein-truncating mutations toward the 3' end of BRCA1, based on nine additional families. PMID- 11927493 TI - Polymorphic CAG/CAA repeat length in the AIB1/SRC-3 gene and prostate cancer risk: a population-based case-control study. AB - In an earlier report, we showed that a shorter CAG repeat length in the androgen receptor (AR) gene is associated with an increased risk of prostate cancer in China, the population with the lowest reported prostate cancer incidence in the world. Because AR coactivators enhance transactivation of AR, in this report we evaluated the relationship of a CAG/CAA repeat length polymorphism in the AIB1/SRC-3 gene (amplified in breast cancer gene 1, a steroid receptor coactivator and an AR coactivator) with prostate cancer risk in a population based case-control study in China. Genomic DNA from 189 prostate cancer patients and 301 healthy controls was used for the PCR-based assay. The AIB1/SRC-3 CAG/CAA repeat length ranged from 24 to 32, with the most common repeat length being 29. Homozygous 29/29 and heterozygous 28/29 were the most common genotypes, with 44 and 30% of the controls harboring these genotypes, respectively. Relative to subjects homozygous for 29 CAG/CAA repeats (29/29 genotype), individuals with the <29/29 genotype had a nonsignificant 31% increased risk [odds ratio (OR), 1.31; 95% confidence interval (CI), 0.87-1.97], whereas those homozygous for the <29 allele had a significant 81% excess risk (OR, 1.81; 95% CI, 1.00-3.28). The combined effect of CAG repeat lengths in the AR and AIB1/SRC-3 genes was also evaluated. Relative to men with both the 29/29 genotype of the AIB1/SRC-3 gene and a long CAG repeat length (> or =23) in the AR gene, those with both the <29/<29 AIB1/SRC-3 genotype and a short CAG repeat length in the AR gene (<23) had a 2.8-fold risk (OR, 2.78; 95% CI, 1.24-6.26). Together, our data indicate that the CAG/CAA repeat length in the AIB1/SRC-3 gene may be associated with prostate cancer risk in Chinese men and that the combination of CAG/CAA repeat lengths in both the AIB1/SRC-3 and AR genes may provide a useful marker for clinically significant prostate cancer. Expanded studies in other populations are needed to confirm this association and the combined effect of AIB1/SRC-3 and other hormone-related genes in prostate cancer etiology. PMID- 11927494 TI - Acquisition and natural history of human papillomavirus type 16 variant infection among a cohort of female university students. AB - Analogous to the epidemiology of different types of human papillomaviruses (HPVs), variants of a single type such as HPV16 may differ in the natural course of infection. A prospective study was conducted in a cohort of female university students who did not have HPV16 DNA or antibody at enrollment. Subjects were followed every 4 months with Pap smear and colposcopic examinations and tests for HPV DNA and antibody. Of 528 women, 62 acquired HPV16 infection during follow-up. The 5-year cumulative incidence was 12.6% for infection with HPV16 prototype-like variants and 3.1% for infection with non-prototype-like (NPL) variants. Among those with incident HPV16 infection, the adjusted odds ratio of detecting NPL variants was 6.0 [95% confidence interval (CI), 1.3-27.5)] for nonwhite women or white women who had had nonwhite sex partners compared with white women who had not had nonwhite sex partners. As compared with women who had not used hormonal contraceptives in the 8 months prior to the first HPV16-positive detection, the adjusted odds ratio of detecting NPL variants increased from 1.6 (95% CI, 0.3 9.1) for those who had used hormonal contraceptives for 1-4 months to 5.4 (95% CI, 1.0-28.3) for those who had used hormonal contraceptives for 5-8 months (P for trend = 0.04). There was no difference in HPV16 seroconversion rates between women with prototype-like variants and women with NPL variants. The increased risk for biopsy-confirmed cervical intraepithelial neoplasia grades 2 or 3 associated with detection of incident infection by NPL variants was not explained by differences in persistence between the two variant groups, indicating that biological mechanisms other than viral persistence may be responsible for the observed difference in risk for cervical intraepithelial neoplasia grades 2 or 3. PMID- 11927495 TI - Human papillomavirus persistence and nutrients involved in the methylation pathway among a cohort of young women. AB - Persistent oncogenic human papillomavirus (HPV) infection is associated with cervical dysplasia. Cofactors, such as nutrient status, may be required for the progression of HPV infection to neoplasia. HPV DNA methylation patterns in vitro have been shown to be associated with viral transcriptional activity. Folate, vitamin B12, vitamin B6, and methionine may function to prevent cervical cancer through their role in DNA methylation. This study was conducted to examine the relationship of dietary intake of folate, vitamin B12, vitamin B6, and methionine, as well as circulating levels of folate and vitamin B12 to HPV persistence. Oncogenic HPV status was determined at baseline and at approximately 3 and 9 months postbaseline. Multivariate logistic regression analysis was used to determine the adjusted odds ratios for persistent HPV infection associated with each tertile of individual nutrient among 201 women with a persistent or intermittent HPV infection. Circulating vitamin B12 levels were inversely associated with HPV persistence (P for trend, 0.037) after adjusting for age, age at first intercourse, marital status, cigarette smoking status, race, and body mass index. In addition, women with circulating levels in the highest tertile (>493.2 pg/ml) of vitamin B12 were less likely to have a persistent infection (adjusted odds ratio = 0.4; 95% confidence interval = 0.17-0.96). No significant associations were observed between HPV persistence and dietary intake of folate, vitamin B12, vitamin B6, or methionine from food alone or from food and supplements combined or from circulating folate. These data suggest a role for circulating vitamin B12 in early cervical carcinogenesis. PMID- 11927496 TI - Steroid hormone levels during pregnancy and incidence of maternal breast cancer. AB - Previous studies evaluating pregnancy hormone levels and maternal breast cancer were limited to surrogate indicators of exposure. This study directly evaluates the association between measured serum steroid hormone levels during pregnancy and maternal risk of breast cancer. A nested case-control study was conducted to examine third-trimester serum levels of total unconjugated estradiol, estrone, estriol, and progesterone in women who were pregnant between 1959 and 1966. Cases (n = 194) were diagnosed with in situ or invasive breast cancer between 1969 and 1991. Controls (n = 374) were matched to cases by age at the time of index pregnancy, using randomized recruitment. Elevated progesterone levels were associated with a decreased incidence of breast cancer [odds ratio (OR) for progesterone > or =270 ng/ml, 0.49; 95% confidence interval (CI), 0.22-1.1] relative to those below the lowest decile. This association was stronger for cancers diagnosed at or before age 50 (OR for progesterone > or =270 ng/ml, 0.3; 95% CI, 0.1-0.9). Increased estrone levels were associated with an increased incidence overall (OR for estrone > or =18.7 ng/ml, 2.5; 95% CI, 1.0-6.2), whereas a positive association with estradiol was not observed. Too few cases occurred within 15 years of the index pregnancy to compare adequately the short- and long-term effects of pregnancy hormone exposure. When estrogen-to progesterone ratios were evaluated, there was an indication of a modest increased incidence of breast cancer for those with high total estrogens and high estrone levels relative to progesterone. These findings suggest that pregnancy steroid hormone levels are risk factors for breast cancer. PMID- 11927497 TI - Eight-year follow-up of the 90,000-person Haimen City cohort: I. Hepatocellular carcinoma mortality, risk factors, and gender differences. AB - In an 8-year follow-up of a prospective cohort study in Haimen City, China, we sought to identify hepatocellular carcinoma (HCC) risk factors in addition to hepatitis B virus (HBV) infection. Two cohorts of adults between ages 25 and 64 years at study entry were followed from 1992-1993 to 2000. The male cohort included 58,545 men, 15.0% of whom were HBV carriers. The female cohort included 25,340 women, 10.7% of whom were HBV carriers. 434,718 person-years of follow-up were accumulated, and 1092 deaths from HCC occurred. The relationship of potential risk factors measured at study entry to HCC mortality was analyzed using Cox proportional hazards models. For males, HCC mortality was significantly associated with HBV infection [relative risk (RR) 18.8; 95% confidence interval (CI), 15.7-22.5], history of acute hepatitis (RR, 2.3; 95% CI, 2.0-2.7), family history of HCC (RR, 2.3; 95% CI, 1.9-2.7), and occupation as a peasant (RR, 1.5; 95% CI, 1.3-1.8). For females, HCC mortality was significantly associated with HBV infection (RR, 33.5; 95% CI, 17.1-65.5) and acute hepatitis history (RR, 4.7; 95% CI, 3.0-7.5). HCC risk was not significantly associated with alcohol consumption, water source, or staple foods in either sex. There was no association with smoking in males, but there was a positive association for females. Environmental and genetic risk factors besides HBV infection play a significant role in HCC mortality in this extremely high-risk population. Gender differences in HCC mortality and known risk factors are substantial and warrant further study. Identification of risk factors amenable to intervention should be a high priority in the prevention of HCC. PMID- 11927499 TI - Colorectal cancer mortality and factors related to the insulin resistance syndrome. AB - It has been proposed that hyperinsulinemia is involved in colon carcinogenesis. An association between post-load plasma glucose (PLG) levels and risk of colorectal cancer mortality would be consistent with this hypothesis. We used data from the Chicago Heart Association Detection Project in Industry to examine the associations of nonfasting PLG and other variables related to the insulin resistance syndrome (i.e., systolic blood pressure, body mass index, uric acid, and resting heart rate) with colorectal cancer mortality. After excluding participants reporting a history of diabetes, 191 and 126 colorectal cancer deaths occurred among 20,433 men and 15,149 women, respectively. In multivariate Cox regression analysis, there was a positive relationship between PLG levels and colorectal cancer mortality for women (P for trend = 0.08) but not for men. When men and women were combined, a trend (P = 0.05) for PLG remained. Examination of clustering of insulin resistance syndrome-related risk factors revealed that men with at least 3 of 4 risk factors (i.e., in the highest quartile of the sex specific distribution for PLG, systolic blood pressure, body mass index, or resting heart rate) had a relative risk (RR) of 1.67 [95% confidence interval (CI), 1.04-2.70] as compared with men who were not in the upper quartile for any of these factors. For women, the RR was 1.29 (95% CI, 0.70-2.37). For men and women combined, the RR was 1.50 (95% CI, 1.03-2.19). These findings provide evidence for a modest association of PLG and insulin resistance syndrome with colorectal cancer mortality and support the insulin hypothesis. PMID- 11927498 TI - CYP2A6 activity determined by caffeine phenotyping: association with colorectal cancer risk. AB - Cytochrome P450 2A6 (CYP2A6) catalyzes the metabolic activation of several procarcinogens including dietary and environmental nitrosamines, and the involvement of CYP2A6 in cancer development has been postulated. CYP2A6 phenotype was determined using caffeine as a probe drug in individuals participating in a case-control study of colorectal cancer (127 cases and 333 controls matched on age, gender, race, and geographic region). Conversion of the caffeine metabolite 1,7-dimethylxanthine (17X) to 1,7-dimethyl uric acid (17U) is catalyzed primarily by CYP2A6, and this activity can be assayed by comparison of urinary molar ratios of metabolites. Caffeine (200 mg) was administered to each participant, and a 4-5 h postadministration urine sample was collected. Urinary metabolites of caffeine were separated by high-performance liquid chromatography and quantified by comparison to authentic standards. We examined the distributions of the ratio, 17U:17X, according to subject characteristics among controls. In case-control comparisons, subjects in the medium and high tertiles of CYP2A6 activity had an increased risk of colorectal cancer compared with subjects with low activity. Odds ratios from a conditional logistic regression model for medium and high 17U:17X ratio were 2.0 (95% confidence interval, 1.1-3.7) and 2.6 (95% confidence interval, 1.5-4.5), respectively (P for trend = 0.001). CYP2A6 phenotype has not been compared previously between cancer cases and controls. We found a strong relationship between CYP2A6 activity, measured by urinary caffeine metabolite ratio, and colorectal cancer risk. PMID- 11927500 TI - Risk of renal and colonic neoplasms and spontaneous pneumothorax in the Birt-Hogg Dube syndrome. AB - The Birt-Hogg-Dube syndrome, a genodermatosis characterized by benign tumors of the hair follicle, has been associated with renal and colonic neoplasms and spontaneous pneumothorax, but the risk of developing these disorders is unknown. We identified risk factors for renal tumors and spontaneous pneumothorax in 98 patients affected with the Birt-Hogg-Dube syndrome, in 13 Birt-Hogg-Dube haplotype carriers, and in 112 unaffected family members. Development of renal tumors was strongly associated with the Birt-Hogg-Dube syndrome and age. The odds ratio for renal tumor in BHD-affected family members adjusted for age was 6.9 (95% confidence interval, 1.5-31.6) and approximately 9.0 for the other risk factors considered. Chromophobe renal carcinoma, an uncommon type of renal cancer, was the predominant type of renal cancer found. Spontaneous pneumothorax was also strongly associated with the Birt-Hogg-Dube syndrome and age. The odds ratio for pneumothorax in BHD-affected individuals, adjusted for age, was 50.3 (95% confidence interval, 6.4-392), and about 32 times higher adjusting for the other risk variables. Colon cancer and colon polyps were not related to the Birt Hogg-Dube syndrome. The Birt-Hogg-Dube syndrome confers an increased risk for the development of renal tumors and spontaneous pneumothorax. We found no increase in risk for the development of colon polyps or colon carcinomas. PMID- 11927501 TI - Allergies and risk of non-Hodgkin's lymphoma by subtype. AB - To investigate the relation between allergy and risk for non-Hodgkin's lymphoma (NHL), data were analyzed from the Selected Cancers Study. Cases (n = 952) were men ages 32-60 years diagnosed with NHL from 1984 to 1988 and identified by eight population-based United States cancer registries. Controls (n = 1691) were recruited by random-digit telephone dialing and frequency matched to cases by age and geographic region of cancer registry. Logistic regression was used to calculate odds ratios and 95% confidence intervals adjusted for age, cancer registry, education, and race/ethnicity. There was no evidence that a general history of allergy was significantly associated with either overall NHL risk (odds ratio: 1.0; 95% confidence interval: 0.8-1.2) or risk for major NHL subtypes (follicular, diffuse, small cell lymphocytic, and immunoblastic). Similarly, no significant associations were observed for the most commonly reported specific allergies, including those to plants, dust, food, animals, and medications. Significant NHL subtype-specific associations were found for allergies to insects (immunoblastic) and chemicals (diffuse and small cell lymphocytic). However, these allergies were reported by relatively few study participants, and the associations may have been because of chance. In conclusion, we found little evidence that either a general history of allergy or commonly reported specific allergies are associated with NHL. PMID- 11927502 TI - Association of the hOGG1 Ser326Cys polymorphism with lung cancer risk. AB - Oxidative stress may be one mechanism by which tobacco smoke causes lung cancer. A common oxidative damage to DNA is the highly mutagenic 7,8-dihydro-8-oxoguanine adduct, which can be repaired by 8-oxoguanine glycosylase I (OGG1). A Ser326Cys substitution polymorphism in the hOGG1 gene has been suggested, based on in vitro data, to reduce the activity of the enzyme. We tested the association of this polymorphism with lung cancer in a population-based, case control study of 298 cases and 405 controls of Caucasian, Japanese, or Native Hawaiian ancestry in Hawaii. Subjects were genotyped with a PCR-RFLP assay, and odds ratios were estimated by logistic regression after adjustment for other observed risk factors, including smoking and vegetable intake. We found marked differences in the frequencies of the hOGG1 Cys variant allele among ethnic groups (45% in Hawaiians, 42% in Japanese, and 22% in Caucasians). The homozygous Cys/Cys genotype was also found to be more common in cases than controls (P = 0.008), with an odds ratio of 2.1 (95% confidence interval: 1.2-3.7) for this genotype compared with the Ser/Ser genotype. Heterozygous individuals were not at increased risk. This association with the Cys/Cys genotype was observed for each sex, ethnic group, and lung cancer cell type. There was also the suggestion that vegetable intake may not be protective against lung cancer among subjects with the Cys/Cys genotype. These data suggest that the presence of two hOGG1 326Cys alleles confers a 2-fold increased risk of lung cancer. Additional studies need to be conducted to confirm this association. PMID- 11927503 TI - The BRCA2 372 HH genotype is associated with risk of breast cancer in Australian women under age 60 years. AB - The BRCA2 N372H nonconservative amino acid substitution polymorphism appears to affect fetal survival in a sex-dependent manner, and the HH genotype was found to be associated with a 1.3-fold risk of breast cancer from pooling five case control studies of Northern European women. We investigated whether the BRCA2 N372H polymorphism was associated with breast cancer in Australian women using a population-based case-control design. The BRCA2 372 genotype was determined in 1397 cases under the age of 60 years at diagnosis of a first primary breast cancer and in 775 population-sampled controls frequency matched for age. Case control analyses and comparisons of genotype distributions were conducted using logistic regression. All of the statistical tests were two-tailed. The HH genotype was independent of age and family history of breast cancer within cases and controls, and was more common in cases (9.2% versus 6.5%). It was associated with an increased risk of breast cancer, 1.47-fold unadjusted (95% confidence interval, 1.05-2.07; P = 0.02), and 1.42-fold (95% confidence interval, 1.00 2.02; P = 0.05) after adjusting for measured risk factors. This effect was still evident after excluding women with any non-Caucasian ancestry or the 33 cases known to have inherited a mutation in BRCA1 or BRCA2, and would explain approximately 3% of breast cancer. The BRCA2 N372H polymorphism appears to be associated with a modest recessively inherited risk of breast cancer in Australian women. This result is consistent with the findings for Northern European women. PMID- 11927504 TI - Steroid 5-alpha reductase type II V89L substitution is not associated with risk of prostate cancer in a multiethnic population study. PMID- 11927505 TI - Human leukocyte antigen class I alleles and cervical neoplasia: no heterozygote advantage. PMID- 11927506 TI - Null association between frequency of cured meat consumption and methylvaline and ethylvaline hemoglobin adduct levels: the N-nitroso brain cancer hypothesis. PMID- 11927507 TI - Correspondence re: Risch, N.: Genetic epidemiology of cancer: interpreting family and twin studies and their implications for molecular genetic approaches. Cancer Epidemiol. Biomark. Prev., 10: 733-741, 2001. PMID- 11927509 TI - The cardiology patient page. Statins: powerful drugs for lowering cholesterol: advice for patients. PMID- 11927510 TI - Myocardial hibernation: restorative or preterminal sleep? PMID- 11927511 TI - Enhanced contractile mechanisms in vasospasm: is endothelial dysfunction the whole story? PMID- 11927512 TI - Does angiotensin-(1-7) contribute to cardiac adaptation and preservation of endothelial function in heart failure? PMID- 11927513 TI - Women, heart failure, and heart failure therapies. PMID- 11927514 TI - Cocaine-related aortic dissection in perspective. PMID- 11927515 TI - Statin attenuates increase in C-reactive protein during estrogen replacement therapy in postmenopausal women. AB - BACKGROUND: HMG-CoA reductase inhibitor (statin) therapy reduces cardiovascular risk, mechanisms of which may include diminished arterial inflammation, as suggested by reduction in levels of C-reactive protein (CRP). Because oral estrogens increase CRP in postmenopausal women, with potential inflammatory and thrombotic consequences that could compromise any benefit to cardiovascular risk, we determined whether the coadministration of a statin might modify the estrogenic effect on CRP. METHODS AND RESULTS: In a double-blind, 3-period crossover study, 28 postmenopausal women (average LDL cholesterol 163+/-36 mg/dL) were randomly assigned to daily conjugated equine estrogens (CEEs) 0.625 mg, simvastatin 10 mg, or their combination for 6 weeks, with each treatment period separated by 6 weeks. CEEs increased median CRP levels from 0.27 to 0.46 mg/dL, simvastatin decreased CRP from 0.29 to 0.28 mg/dL, and the therapies combined increased CRP from 0.28 to 0.36 mg/dL (all P< or =0.02 versus respective baseline values). Post hoc testing showed that the 29% increase in CRP on the combination of CEEs with simvastatin was significantly less than the 70% increase in CRP on CEEs alone (P<0.05). The effect of combination therapy on CRP levels did not correlate with baseline CRP or with baseline or treatment-induced changes in levels of interleukin-6, lipoproteins, or flow-mediated dilation of the brachial artery as a measure of nitric oxide bioactivity. CONCLUSIONS: The combination of statin with estrogen may attenuate the potential harmful effects of estrogen therapy in postmenopausal women and maximize any benefit to cardiovascular risk. PMID- 11927516 TI - Inhalation of fine particulate air pollution and ozone causes acute arterial vasoconstriction in healthy adults. AB - BACKGROUND: Fine particulate air pollution and ozone are associated with increased cardiovascular events. To help explain the mechanism behind these observations, we investigated the effect of air pollution exposure on vascular function. METHODS AND RESULTS: Twenty-five healthy adults underwent a randomized, double-blind, crossover study comparing the vascular response to the 2-hour inhalation of approximately 150 microg/m(3) of concentrated ambient fine particles (CAP) plus ozone (120 ppb) versus the response to the inhalation of filtered air. High-resolution vascular ultrasonography was used to measure alterations in brachial artery diameter, endothelial-dependent flow-mediated dilatation (FMD) and endothelial-independent nitroglycerin-mediated dilatation (NMD). Exposure to CAP plus ozone caused a significant brachial artery vasoconstriction compared with filtered air inhalation (-0.09+/-0.15 mm versus +0.01+/-0.18 mm, P=0.03). There were no significant differences in FMD (+0.29+/ 4.11% versus -0.03+/-6.63%, P=0.88), NMD (+3.87+/-5.43% versus +3.46+/-7.92%, P=0.83), or blood pressure responses between exposures. CONCLUSIONS: Short-term inhalation of fine particulate air pollution and ozone at concentrations that occur in the urban environment causes acute conduit artery vasoconstriction. PMID- 11927517 TI - Increased myocardial gene expression of tumor necrosis factor-alpha and nitric oxide synthase-2: a potential mechanism for depressed myocardial function in hibernating myocardium in humans. AB - BACKGROUND: Whether cardioinhibitory cytokines are elevated in regions of hibernating myocardium and account in part for the depression in resting function is currently not known. Methods and Results- Thirteen patients with stable ischemic ventricular dysfunction scheduled for bypass surgery underwent preoperative dobutamine echocardiography (DE) and intraoperative myocardial biopsies. The numbers of copies of mRNA for the negatively inotropic cytokines tumor necrosis factor-alpha (TNF-alpha) and inducible nitric oxide synthase (NOS2) were quantified by reverse transcription-polymerase chain reaction. In normal segments, myocardial TNF-alpha was barely detectable (1.2+/-0.4 copies per 10(6) copies of beta-actin). A 13.7-fold increase in myocardial TNF-alpha was observed in dysfunctional segments with a biphasic response to DE (contractile reserve and ischemia) and was highest (45.5-fold) in segments with ischemia and without contractile reserve (P<0.001). A similar graded increase was seen for NOS2. Cytokine results were also similar if analysis was performed using recovery of function at 3 months as the index of viability. The change in serum TNF-alpha and nitrite levels from baseline to 3 months after surgery correlated inversely with both the change in ejection fraction and the number of DE viable segments (r=-0.92 to -0.93; P<0.001). CONCLUSIONS: TNF-alpha and NOS2 gene expression is regionally upregulated in hibernating myocardium to a level intermediate between that of normal regions and ischemic regions without contractile reserve. This, along with a decline in serum cytokine levels after revascularization proportional to the extent of myocardial viability, suggests a contributing role for cardioinhibitory cytokines in the observed depression of function seen in hibernating myocardium. PMID- 11927518 TI - Endothelial cell senescence in human atherosclerosis: role of telomere in endothelial dysfunction. AB - BACKGROUND: The functional changes associated with cellular senescence may be involved in human aging and age-related vascular disorders. We have shown the important role of telomere and telomerase in vascular cell senescence in vitro. Progressive telomere shortening in vivo has been observed in the regions susceptible to atherosclerosis, implying contributions to atherogenesis. However, whether senescent vascular cells are present in the vasculature and contribute to the pathogenesis of atherosclerosis remains unclear. METHODS AND RESULTS: Senescence-associated beta-galactosidase (beta-gal) activity was examined in the coronary arteries and the internal mammary arteries retrieved from autopsied individuals who had had ischemic heart diseases. Strong beta-gal stainings were observed in atherosclerotic lesions of the coronary arteries but not in the internal mammary arteries. An immunohistochemical analysis using anti-factor VIII antibody demonstrated that beta-gal stained cells are vascular endothelial cells. To determine whether endothelial cell senescence causes endothelial dysfunction, we induced senescence in human aortic endothelial cells (HAECs) by inhibiting telomere function and examined the expression of intercellular adhesion molecule (ICAM)-1 and endothelial nitric oxide synthase (eNOS) activity. Senescent HAECs exhibited increased ICAM-1 expression and decreased eNOS activity, both of which are alterations implicated in atherogenesis. In contrast, introduction of telomerase catalytic component significantly extended the life span and inhibited the functional alterations associated with senescence in HAECs. CONCLUSIONS: Vascular endothelial cells with senescence-associated phenotypes are present in human atherosclerotic lesions, and endothelial cell senescence induced by telomere shortening may contribute to atherogenesis. PMID- 11927519 TI - Suppression of coronary artery spasm by the Rho-kinase inhibitor fasudil in patients with vasospastic angina. AB - BACKGROUND: Increased activity of Rho-kinase causes hypercontraction of vascular smooth muscle and has been implicated as playing a pathogenetic role in divergent cardiovascular diseases such as coronary artery spasm. We examined whether an intracoronary infusion of fasudil, a selective Rho-kinase inhibitor, would attenuate coronary vasoconstrictor responses to acetylcholine (ACh) in patients with vasospastic angina. METHODS AND RESULTS: We studied 20 consecutive patients in whom coronary artery spasm was provoked by intracoronary ACh. The patients underwent a second ACh challenge after pretreatment with intracoronary saline (n=5) or fasudil (n=15; 300 microg/min for 15 minutes). Angina and coronary vasospasm were reproducibly induced by the second testing in patients who received saline. In contrast, fasudil markedly attenuated the coronary constriction induced by ACh (P<0.001) and prevented the occurrence of chest pain and ischemic ECG changes in all treated patients (both P<0.01 versus saline). Fasudil, at the dose used in this study, did not significantly change systemic hemodynamics or baseline coronary blood flow. CONCLUSIONS: Fasudil was effective in preventing ACh-induced coronary artery spasm and resultant myocardial ischemia in patients with vasospastic angina. We suggest that this Rho-kinase inhibitor may be a novel therapeutic intervention to treat ischemic coronary syndromes caused by coronary artery spasm. PMID- 11927520 TI - Angiotensin-(1-7) attenuates the development of heart failure after myocardial infarction in rats. AB - BACKGROUND: The renin-angiotensin system (RAS) is a key player in the progression of heart failure. Angiotensin-(1-7) is thought to modulate the activity of the RAS. Furthermore, this peptide may play a part in the beneficial effects of angiotensin-converting enzyme inhibitors in cardiovascular disease. We assessed the effects of angiotensin-(1-7) on the progression of heart failure. METHODS AND RESULTS: Male Sprague-Dawley rats underwent either coronary ligation or sham surgery. Two weeks after induction of myocardial infarction, intravenous infusion of angiotensin-(1-7) (24 microg/kg per hour) or saline was started by minipump. After 8 weeks of treatment, hemodynamic parameters were measured, endothelial function was assessed in isolated aortic rings, and plasma angiotensin-(1-7) levels were determined. Myocardial infarction resulted in a significant deterioration of left ventricular systolic and diastolic pressure, dP/dt, and coronary flow. Raising plasma levels 40-fold, angiotensin-(1-7) infusion attenuated this impairment to a nonsignificant level, markedly illustrated by a 40% reduction in left ventricular end-diastolic pressure. Furthermore, angiotensin-(1-7) completely preserved aortic endothelial function, whereas endothelium-dependent relaxation in aortas of saline-treated infarcted rats was significantly decreased. CONCLUSIONS: Angiotensin-(1-7) preserved cardiac function, coronary perfusion, and aortic endothelial function in a rat model for heart failure. PMID- 11927521 TI - Modulation of anthracycline-induced myofibrillar disarray in rat ventricular myocytes by neuregulin-1beta and anti-erbB2: potential mechanism for trastuzumab induced cardiotoxicity. AB - BACKGROUND: There is an increased incidence of heart failure in patients treated concurrently with anthracyclines and the chemotherapeutic anti-erbB2 agent trastuzumab (Herceptin). On the basis of our previous studies with recombinant neuregulin-1beta (NRG-1beta), a ligand for the erbB2 receptor tyrosine kinase, we hypothesized that activation of erbB2 by anti-erbB2 versus NRG-1 would cause differential effects on myocyte intracellular signaling as well as anthracycline induced myofibrillar injury and might potentially account for the clinical toxicity of trastuzumab in the setting of concurrent anthracycline therapy. METHODS AND RESULTS: We tested this hypothesis using adult rat ventricular myocytes (ARVMs) in culture, assessing myofibrillar structure by immunostaining for myomesin and filamentous actin. Activation of erbB2, extracellular signal regulated kinase 1/2 (Erk1/2), and Akt was assessed by use of antibodies to phosphorylated activated receptor or kinase detected by immunoblot. ARVMs treated with doxorubicin (0.1 to 0.5 micromol/L) showed a concentration-dependent increase in myofilament disarray. NRG-1beta (10 ng/mL) activated erbB2, Erk1/2, and Akt in ARVMs and significantly reduced anthracycline-induced disarray. In contrast to NRG-1beta, anti-erbB2 (1 microg/mL) caused rapid phosphorylation of erbB2 but not Erk1/2 or Akt, with downregulation of erbB2 by 24 hours. Concomitant treatment of myocytes with anti-erbB2 and doxorubicin caused a significant increase in myofibrillar disarray versus doxorubicin alone. CONCLUSIONS: NRG-1beta/erbB signaling regulates anthracycline-induced myofilament injury. The increased susceptibility of myofilaments to doxorubicin in the presence of antibody to erbB2 may explain the contractile dysfunction seen in patients receiving concurrent trastuzumab and anthracyclines. PMID- 11927522 TI - Effect of 3 months of antimicrobial treatment with clarithromycin in acute non-q wave coronary syndrome. AB - BACKGROUND: Coronary artery disease, an inflammatory disease, may be caused by infection. We investigated whether the antibiotic clarithromycin would reduce morbidity and mortality in patients with acute non-Q-wave coronary syndrome. METHODS AND RESULTS: Altogether, 148 patients with acute non-Q-wave infarction or unstable angina were randomly assigned to receive double-blind treatment with either clarithromycin or placebo for 3 months. The primary end point was a composite of death, myocardial infarction, or unstable angina during treatment; the secondary end point was occurrence of any cardiovascular event during the entire follow-up period (average 555 days, range 138 to 924 days). There was a trend toward fewer patients meeting primary end-point criteria in the clarithromycin group than in the placebo group (11 versus 19 patients, respectively; risk ratio 0.54, 95% CI 0.25 to 1.14; P=0.10). By the end of the entire follow-up, 16 patients in the clarithromycin group and 27 in the placebo group had experienced a cardiovascular event (risk ratio 0.49, 95% CI 0.26 to 0.92; P=0.03). CONCLUSIONS: Clarithromycin appears to reduce the risk of ischemic cardiovascular events in patients presenting with acute non-Q-wave infarction or unstable angina. No signs of this effect diminishing were observed during follow up. PMID- 11927523 TI - Expression of macrophage migration inhibitory factor in different stages of human atherosclerosis. AB - BACKGROUND: Atherosclerosis is a chronic inflammatory response of the arterial wall to injury. Macrophage migration inhibitory factor (MIF), a cytokine with potent inflammatory functions, was thus considered to be important in atherosclerotic lesion evolution. METHODS AND RESULTS: We studied the presence and distribution of MIF immunoreactivity (MIF-IR) and MIF mRNA in internal mammary arteries with a normal histology and arteries with plaques in different stages of human atherosclerosis. To address a potential role for the coactivator Jab1 as a cellular mediator of MIF effects in vascular tissue, we correlated the expression of MIF to that of Jab1 by using immunohistochemistry and coimmunoprecipitation. We further sought to determine a potential functional role for endothelium-derived MIF in early atherogenesis by studying the effects of oxidized LDL on MIF expression in cultured human umbilical vascular endothelial cells. The results showed that MIF-IR and Jab1-IR are found in all cell types present in atherosclerotic lesions, that MIF-IR is upregulated during progression of atherosclerosis, that MIF is produced locally in the arterial wall, and that all MIF(+) cells are simultaneously Jab1(+). Coimmunoprecipitation experiments demonstrated in vivo complex formation between MIF and Jab1 in plaques. MIF expression in human umbilical vascular endothelial cells and a macrophage line was upregulated after stimulation with oxidized LDL. CONCLUSIONS: MIF is produced abundantly by various cells in all types of human atherosclerotic lesions and thus may play an important role in early plaque development and advanced complicated lesions. MIF-Jab1 complexes could serve critical regulatory functions in atherosclerotic lesion evolution. PMID- 11927525 TI - Coronary flow velocity reserve after percutaneous interventions is predictive of periprocedural outcome. AB - BACKGROUND: Because heterogeneous results have been reported, we assessed coronary flow velocity changes in individuals who underwent percutaneous transluminal coronary angioplasty (PTCA) and examined their impact on clinical outcome. METHODS AND RESULTS: As part of the Doppler Endpoints Balloon Angioplasty Trial Europe (DEBATE) II study, 379 patients underwent Doppler flow guided angioplasty. All patients were evaluated according to their coronary flow velocity reserve (CFVR) results (> or =2.5 or < 2.5) at the end of the procedure. A CFVR < 2.5 after angioplasty was associated with an elevated baseline blood flow velocity in both the target artery and reference artery. CFVR before PTCA and CFVR in the reference artery were independent predictors of an optimal CFVR after balloon angioplasty (CFVR before PTCA: odds ratio [OR], 2.26; 95% confidence interval [CI], 1.57 to 3.24; CFVR in reference artery: OR, 1.90; 95% CI, 1.21 to 2.98; both P<0.001) and stent implantation (before PTCA: OR, 2.54; 95% CI, 1.47 to 4.36; reference artery: OR, 1.97; 95% CI, 1.07 to 3.87; both P<0.05). A low CFVR at the end of the procedure was an independent predictor of major adverse cardiac events (MACE) at 30 days (OR, 4.71; 95% CI, 1.14 to 25.92; P=0.034) and at 1 year (OR, 2.06; 95% CI, 1.16 to 3.66; P=0.014). After excluding MACE at 30 days, no difference in MACE at 1 year was observed between the patients with and without a CFVR < 2.5 at the end of the procedure. CONCLUSIONS: A low postprocedural CFVR was associated with a worse periprocedural outcome (which was related to microcirculatory disturbances), but there was no significant difference at late follow-up. PMID- 11927524 TI - Risk stratification for postoperative cardiovascular events via noninvasive assessment of endothelial function: a prospective study. AB - BACKGROUND: Brachial artery endothelial function is impaired in individuals with atherosclerosis and coronary risk factors and improves with risk reduction therapy. However, the predictive value of brachial artery endothelial dysfunction for future cardiovascular events is unknown. METHODS AND RESULTS: We preoperatively examined brachial artery vasodilation using ultrasound in 187 patients undergoing vascular surgery. Patients were prospectively followed for 30 days after surgery. Forty-five patients had a postoperative event, including cardiac death (3), myocardial infarction (12), unstable angina/ischemic ventricular fibrillation (2), stroke (3), or elevated troponin I, reflecting myocardial necrosis (25). Preoperative endothelium-dependent flow-mediated dilation was significantly lower in patients with an event (4.9+/-3.1%) than in those without an event (7.3+/-5%; P<0.001), whereas endothelium-independent vasodilation to nitroglycerin was similar in both groups. In a Cox proportional hazards model, the independent predictors of events were age (P=0.001), renal insufficiency (P=0.03), noncarotid surgery (P=0.05), and lower brachial artery flow-mediated dilation (P=0.007). If troponin I elevation was not considered an event, low flow-mediated dilation remained an independent predictor of risk (odds ratio 9.0, 95% CI 1.2 to 68; P=0.03). When a flow-mediated dilation cutpoint of 8.1% was used, endothelial function had a sensitivity of 95%, specificity of 37%, and negative predictive value of 98% for events. CONCLUSIONS: Impaired brachial artery endothelial function independently predicts postoperative cardiac events, which supports a role for endothelial dysfunction in the pathogenesis of cardiovascular disease. The strong negative predictive value of preserved endothelial function raises the possibility that assessment of brachial artery flow-mediated dilation will be useful in the management of patients undergoing vascular surgery. PMID- 11927526 TI - Fragment reconstruction of coronary arteries by transesophageal echocardiography: a method for visualizing coronary arteries with ultrasound. AB - BACKGROUND: If transesophageal echocardiography (TEE) is to play a role in coronary diagnostics, satisfactory image documentation of the coronary status is indispensable so that the requirements for validity and quality assurance of a medical examination can be met. Our goal was to develop a suitable and valid procedure for imaging coronary arteries with 2D TEE. METHODS AND RESULTS: After pilot trials and formulation of requirements, a new method of imaging coronary arteries was designed, supported by theoretical and mathematical principles: FRC TEE, or fragment reconstruction of coronary arteries by means of 2D TEE. The method generates images of successive vessel fragments so that reconstruction in a summation picture is possible. The procedure orients itself to the vessel and ensures proof of identity by permanently following it on the screen. FRC-TEE was evaluated in 12 consecutive patients with an indication for TEE. One hundred percent visualization in proximal, middle, and distal segments was achieved. The total lengths visualized were 10.74+/-2.56 cm for the right and 8.67+/-1.12 cm for the left coronary artery. Stents and stenoses within the vessels were identified convincingly. Because of a technical problem, investigation of the left anterior descending artery was initially avoided. In a pilot trial series (n=12), 6.65+/-0.92 cm of this vessel was imaged. CONCLUSIONS: FRC-TEE is a newly developed method of visualizing coronary arteries by means of ultrasound, which permits good pictorial documentation. Initial studies have achieved promising results. Representation of the length of coronary arteries with ultrasound was considerably improved with this technique. PMID- 11927527 TI - Metoprolol CR/XL in female patients with heart failure: analysis of the experience in Metoprolol Extended-Release Randomized Intervention Trial in Heart Failure (MERIT-HF). AB - BACKGROUND: Underrepresentation of women in heart failure clinical trials has limited conclusions regarding the effect of various management strategies on survival in women with heart failure and decreased left ventricular ejection fraction (LVEF). METHODS AND RESULTS: MERIT-HF (Metoprolol Extended-Release Randomized Intervention Trial in Heart Failure) was a randomized, placebo controlled study, the purpose of which was to evaluate the effect of metoprolol controlled-release/extended-release (CR/XL) in 3991 patients with New York Heart Association class II to IV heart failure and LVEF < or =0.40. We performed a post hoc analysis to evaluate the effect of metoprolol CR/XL on outcome in women (n=898), including the outcome in 183 women with severe heart failure (New York Heart Association class III/IV and LVEF < 0.25). Treatment with metoprolol CR/XL in women resulted in a 21% reduction in the primary combined end point of all cause mortality/all-cause hospitalizations (164 versus 137 patients; P=0.044). The number of cardiovascular hospitalizations was reduced by 29% (164 versus 120; P=0.013), and hospitalization for worsening heart failure was reduced by 42% (95 versus 56; P=0.021). Similar results were noted in the subgroup of women with severe heart failure, with a 57% reduction in cardiovascular hospitalizations (63 versus 30; P=0.005) and a 72% reduction in hospitalization due to worsening heart failure (46 versus 14; P=0.0004). A pooling of mortality results from MERIT-HF, the Cardiac Insufficiency Bisoprolol Study (CIBIS II), and the Carvedilol Prospective Randomized Cumulative Survival Study (COPERNICUS) showed very similar survival benefits in women and men. CONCLUSIONS: The beneficial effects of metoprolol CR/XL extend to women with heart failure, including women with clinically stable severe heart failure. PMID- 11927528 TI - Acute aortic dissection related to crack cocaine. AB - BACKGROUND: Although single case reports have described acute aortic dissection in relation to cocaine use, this condition is not widely recognized, and the features of cocaine-related aortic dissection have not been defined. METHODS AND RESULTS: We reviewed all available hospital charts from 1981 to 2001 with the ICD 9 diagnosis of aortic dissection. Among the 38 cases of acute aortic dissection, 14 (37%) were related to cocaine use. Crack cocaine was smoked in 13 cases and powder cocaine was snorted in 1 case. The mean interval between cocaine use and the onset of symptoms was 12 hours (range, 0 to 24). Patients with cocaine related dissection were much younger and more likely to undergo surgery compared with patients with aortic dissection without cocaine use. Most in the cocaine group were black, with a history of untreated hypertension. However, the two groups did not differ in other respects, including dissection type. CONCLUSIONS: In an inner city population, acute aortic dissection in the setting of crack cocaine use is common, presumably as a consequence of abrupt, transient, severe hypertension and catecholamine release. This diagnosis should be considered in cocaine users with severe chest pain. PMID- 11927529 TI - In vivo temperature heterogeneity of atherosclerotic plaques is determined by plaque composition. AB - BACKGROUND: Temperature heterogeneity of atherosclerotic plaques has been associated with macrophage accumulation in ex vivo studies. We investigated in vivo whether modifying the cell composition of rabbit atherosclerotic plaques by dietary cholesterol lowering can influence temperature heterogeneity. METHODS AND RESULTS: Twenty New Zealand rabbits were randomized to either a normal (n=10) or cholesterol-rich (0.3%) diet (n=10) for 6 months. Thereafter, intravascular ultrasound and intravascular catheter-based thermography of the surface of aortic arch and descending aorta were performed in all animals. Ten control and 5 hypercholesterolemic rabbits were euthanized, and their aortas were analyzed histologically. The 5 remaining rabbits received a normal diet for 3 months and underwent repeat ultrasound and thermography before euthanasia followed by histology. Ex vivo temperature was measured in 3 additional rabbits at 6 months to correlate local temperature with local plaque composition. In control animals, plaque formation and temperature heterogeneity were absent. In hypercholesterolemic rabbits, plaque formation was prominent in the thoracic aorta. Plaques were composed of fibromuscular tissue and contained, underneath endothelial cells, an accumulation of foam cells of macrophage origin. Temperature heterogeneity was markedly elevated and increased with plaque thickness. Importantly, after 3 months of cholesterol lowering, plaque thickness remained unchanged, but temperature heterogeneity was significantly decreased. This paralleled plaque histology, which showed a marked loss of macrophages. The ex vivo experiments demonstrated the relation between local temperature and local total macrophage mass. CONCLUSIONS: In vivo temperature heterogeneity of rabbit atherosclerotic plaques is determined by plaque composition. In vivo thermography may have important clinical implications in the assessment of plaque composition. PMID- 11927530 TI - Tissue Doppler imaging differentiates physiological from pathological pressure overload left ventricular hypertrophy in rats. AB - BACKGROUND: The myocardial velocity gradient (MVG) is a recent index of regional myocardial function derived from endocardial and epicardial velocities obtained by tissue Doppler imaging (TDI). This index might be useful for discriminating between physiological and pathological left ventricular hypertrophy (LVH) and for documenting the early transition from compensated LVH to heart failure. We sought to compare MVG measured across the left ventricular posterior wall between normal rats and rats with physiological (exercise) and pathological (pressure-overload) LVH. METHODS AND RESULTS: Wistar rats were assigned to one of the following groups: sedentary, exercise (swimming), and 2-month or 9-month abdominal aortic banding. Compared with sedentary rats, exercise and 2-month banding led to similar and significant LVH. After 2-month banding, conventional parameters of systolic function (left ventricular fractional shortening and dP/dt(max)) were not affected. However, systolic and diastolic MVG were similar in exercise and sedentary rats but were significantly lower in rats with aortic banding. Aortic debanding after 2 months led to a full recovery of MVG, whereas MVG remained decreased when debanding was performed after 9 months. CONCLUSIONS: Myocardial contraction and relaxation assessed by TDI were impaired in pressure-overload LVH but not in exercise LVH. Therefore, TDI is more sensitive than conventional echocardiography for assessing myocardial dysfunction in pressure-overload LVH and for predicting early recovery in myocardial function after loading conditions normalization. PMID- 11927531 TI - Suppression of murine cardiac allograft arteriopathy by long-term blockade of CD40-CD154 interactions. AB - BACKGROUND: The interaction between CD40 on antigen-presenting cells and CD40L on T cells is critical in allograft rejection. CD154 blockade suppresses allograft rejection, but the role of this pathway in allograft vasculopathy remains obscure. METHODS AND RESULTS: A vascularized murine heterotopic cardiac transplant model was used to test whether perioperative CD154 blockade suppresses allograft vasculopathy or whether long-term CD154 blockade is required to suppress allograft vasculopathy. Perioperative CD154 blockade consisted of MR1 given on days -1, 1, and 3; long-term blockade consisted of MR1 given on days -1, 1, and 3 and continued twice weekly for 8 weeks. Allografts treated with perioperative or long-term CD154 blockade survived indefinitely. Perioperative and long-term treatment with control antibody (Ha4/8) resulted in uniform early rejection. Perioperative CD154 blockade transiently reduced early T-cell and macrophage infiltration in parallel with a transient reduction in endothelial adhesion receptor expression. Although perioperative CD154 blockade prevented allograft failure, it did not reduce allograft vasculopathy; mean neointimal cross-sectional area in perioperative MR1-treated and Ha4/8-treated recipients was 43+/-7% and 50+/-12%, respectively (P=NS). In contrast, mean neointimal cross sectional area in long-term, MR1-treated recipients was 19+/-3% (P<0.001 versus perioperative MR1). Long-term CD154 blockade also suppressed endothelial E selectin, P-selectin, and intracellular adhesion molecule-1 expression and improved graft function 3.5-fold versus control (P<0.05). CONCLUSIONS: These data show that perioperative CD154 blockade mitigates acute rejection but long-term CD154 blockade may result in decreased allograft endothelial activation and is required to suppress allograft arteriopathy. PMID- 11927532 TI - Images in cardiovascular medicine: Hypothenar hammer syndrome. PMID- 11927533 TI - Images in cardiovascular medicine. Unusual angiographic appearance of scimitar syndrome associated with primitive hepatic venous plexus. PMID- 11927535 TI - An essential role for Prox1 in the induction of the lymphatic endothelial cell phenotype. AB - The process of angiogenesis has been well documented, but little is known about the biology of lymphatic endothelial cells and the molecular mechanisms controlling lymphangiogenesis. The homeobox gene Prox1 is expressed in a subpopulation of endothelial cells that, after budding from veins, gives rise to the mammalian lymphatic system. In Prox1(-)(/-) embryos, this budding becomes arrested at around embryonic day (E)11.5, resulting in embryos without lymphatic vasculature. Unlike the endothelial cells that bud off in E11.5 wild-type embryos, those of Prox1-null embryos did not co-express any lymphatic markers such as VEGFR-3, LYVE-1 or SLC. Instead, the mutant cells appeared to have a blood vascular phenotype, as determined by their expression of laminin and CD34. These results suggest that Prox1 activity is required for both maintenance of the budding of the venous endothelial cells and differentiation toward the lymphatic phenotype. On the basis of our findings, we propose that a blood vascular phenotype is the default fate of budding embryonic venous endothelial cells; upon expression of Prox1, these budding cells adopt a lymphatic vasculature phenotype. PMID- 11927536 TI - The novel product of a five-exon stargazin-related gene abolishes Ca(V)2.2 calcium channel expression. AB - We have cloned and characterized a new member of the voltage-dependent Ca(2+) channel gamma subunit family, with a novel gene structure and striking properties. Unlike the genes of other potential gamma subunits identified by their homology to the stargazin gene, CACNG7 is a five-, and not four-exon gene whose mRNA encodes a protein we have designated gamma(7). Expression of human gamma(7) has been localized specifically to brain. N-type current through Ca(V)2.2 channels was almost abolished when co-expressed transiently with gamma(7) in either Xenopus oocytes or COS-7 cells. Furthermore, immunocytochemistry and western blots show that gamma(7) has this effect by causing a large reduction in expression of Ca(V)2.2 rather than by interfering with trafficking or biophysical properties of the channel. No effect of transiently expressed gamma(7) was observed on pre-existing endogenous N-type calcium channels in sympathetic neurones. Low homology to the stargazin-like gamma subunits, different gene structure and the unique functional properties of gamma(7) imply that it represents a distinct subdivision of the family of proteins identified by their structural and sequence homology to stargazin. PMID- 11927537 TI - CLAC: a novel Alzheimer amyloid plaque component derived from a transmembrane precursor, CLAC-P/collagen type XXV. AB - We raised monoclonal antibodies against senile plaque (SP) amyloid and obtained a clone 9D2, which labeled amyloid fibrils in SPs and reacted with approximately 50/100 kDa polypeptides in Alzheimer's disease (AD) brains. We purified the 9D2 antigens and cloned a cDNA encoding its precursor, which was a novel type II transmembrane protein specifically expressed in neurons. This precursor harbored three collagen-like Gly-X-Y repeat motifs and was partially homologous to collagen type XIII. Thus, we named the 9D2 antigen as CLAC (collagen-like Alzheimer amyloid plaque component), and its precursor as CLAC-P/collagen type XXV. The extracellular domain of CLAC-P/collagen type XXV was secreted by furin convertase, and the N-terminus of CLAC deposited in AD brains was pyroglutamate modified. Both secreted and membrane-tethered forms of CLAC-P/collagen type XXV specifically bound to fibrillized Abeta, implicating these proteins in beta amyloidogenesis and neuronal degeneration in AD. PMID- 11927538 TI - Lack of collagen XVIII/endostatin results in eye abnormalities. AB - Mice lacking collagen XVIII and its proteolytically derived product endostatin show delayed regression of blood vessels in the vitreous along the surface of the retina after birth and lack of or abnormal outgrowth of retinal vessels. This suggests that collagen XVIII/endostatin is critical for normal blood vessel formation in the eye. All basement membranes in wild-type eyes, except Descemet's membrane, showed immunogold labeling with antibodies against collagen XVIII. Labeling at sites where collagen fibrils in the vitreous are connected with the inner limiting membrane and separation of the vitreal matrix from the inner limiting membrane in mutant mice indicate that collagen XVIII is important for anchoring vitreal collagen fibrils to the inner limiting membrane. The findings provide an explanation for high myopia, vitreoretinal degeneration and retinal detachment seen in patients with Knobloch syndrome caused by loss-of-function mutations in collagen XVIII. PMID- 11927539 TI - GCAP1 rescues rod photoreceptor response in GCAP1/GCAP2 knockout mice. AB - Visual transduction in retinal photoreceptors operates through a dynamic interplay of two second messengers, Ca(2+) and cGMP. Ca(2+) regulates the activity of guanylate cyclase (GC) and the synthesis of cGMP by acting on a GC activating protein (GCAP). While this action is critical for rapid termination of the light response, the GCAP responsible has not been identified. To test if GCAP1, one of two GCAPs present in mouse rods, supports the generation of normal flash responses, transgenic mice were generated that express only GCAP1 under the control of the endogenous promoter. Paired flash responses revealed a correlation between the degree of recovery of the rod a-wave and expression levels of GCAP1. In single cell recordings, the majority of the rods generated flash responses that were indistinguishable from wild type. These results demonstrate that GCAP1 at near normal levels supports the generation of wild-type flash responses in the absence of GCAP2. PMID- 11927540 TI - Dual roles for the Dab2 adaptor protein in embryonic development and kidney transport. AB - The Disabled-2 (Dab2) gene has been proposed to act as a tumor suppressor. Cell culture studies have implicated Dab2 in signal transduction by mitogens, TGFbeta and endocytosis of lipoprotein receptors. To identify in vivo functions of Dab2, targeted mutations were made in the mouse. In the absence of Dab2, embryos arrest prior to gastrulation with a phenotype reminiscent of those caused by deletion of some TGFbeta signal transduction molecules involved in Nodal signaling. Dab2 is expressed in the extra-embryonic visceral endoderm but not in the epiblast. Dab2 could be conditionally deleted from the embryo without affecting normal development, showing that Dab2 is required in the visceral endoderm but dispensable in the embryo proper. Conditionally mutant Dab2(-/-) mice are overtly normal, but have reduced clathrin-coated pits in kidney proximal tubule cells and excrete specific plasma proteins in the urine, consistent with reduced transport by a lipoprotein receptor, megalin/gp330, in the proximal tubule. This evidence indicates that Dab2 is pleiotropic and regulates both visceral endoderm function and lipoprotein receptor trafficking in vivo. PMID- 11927541 TI - A positive feedback loop stabilizes the guanine-nucleotide exchange factor Cdc24 at sites of polarization. AB - In Saccharomyces cerevisiae, activation of Cdc42 by its guanine-nucleotide exchange factor Cdc24 triggers polarization of the actin cytoskeleton at bud emergence and in response to mating pheromones. The adaptor protein Bem1 localizes to sites of polarized growth where it interacts with Cdc42, Cdc24 and the PAK-like kinase Cla4. We have isolated Bem1 mutants (Bem1-m), which are specifically defective for binding to Cdc24. The mutations map within the conserved PB1 domain, which is necessary and sufficient to interact with the octicos peptide repeat (OPR) motif of Cdc24. Although Bem1-m mutant proteins localize normally, bem1-m cells are unable to maintain Cdc24 at sites of polarized growth. As a consequence, they are defective for apical bud growth and the formation of mating projections. Localization of Bem1 to the incipient bud site requires activated Cdc42, and conversely, expression of Cdc42-GTP is sufficient to accumulate Bem1 at the plasma membrane. Thus, our results suggest that Bem1 functions in a positive feedback loop: local activation of Cdc24 produces Cdc42-GTP, which recruits Bem1. In turn, Bem1 stabilizes Cdc24 at the site of polarization, leading to apical growth. PMID- 11927542 TI - Intramembrane cleavage of microneme proteins at the surface of the apicomplexan parasite Toxoplasma gondii. AB - Apicomplexan parasites actively secrete proteins at their apical pole as part of the host cell invasion process. The adhesive micronemal proteins are involved in the recognition of host cell receptors. Redistribution of these receptor-ligand complexes toward the posterior pole of the parasites is powered by the actomyosin system of the parasite and is presumed to drive parasite gliding motility and host cell penetration. The microneme protein protease termed MPP1 is responsible for the removal of the C-terminal domain of TgMIC2 and for shedding of the protein during invasion. In this study, we used site-specific mutagenesis to determine the amino acids essential for this cleavage to occur. Mapping of the cleavage site on TgMIC6 established that this processing occurs within the membrane-spanning domain, at a site that is conserved throughout all apicomplexan microneme proteins. The fusion of the surface antigen SAG1 with these transmembrane domains excluded any significant role for the ectodomain in the cleavage site recognition and provided evidence that MPP1 is constitutively active at the surface of the parasites, ready to sustain invasion at any time. PMID- 11927543 TI - Levels of circumsporozoite protein in the Plasmodium oocyst determine sporozoite morphology. AB - The sporozoite stage of the Plasmodium parasite is formed by budding from a multinucleate oocyst in the mosquito midgut. During their life, sporozoites must infect the salivary glands of the mosquito vector and the liver of the mammalian host; both events depend on the major sporozoite surface protein, the circumsporozoite protein (CS). We previously reported that Plasmodium berghei oocysts in which the CS gene is inactivated do not form sporozoites. Here, we analyzed the ultrastructure of P.berghei oocyst differentiation in the wild type, recombinants that do not produce or produce reduced amounts of CS, and corresponding complemented clones. The results indicate that CS is essential for establishing polarity in the oocyst. The amounts of CS protein correlate with the extent of development of the inner membranes and associated microtubules underneath the oocyst outer membrane, which normally demarcate focal budding sites. This is a first example of a protein controlling both morphogenesis and infectivity of a parasite stage. PMID- 11927544 TI - Plasmodium sporozoite invasion into insect and mammalian cells is directed by the same dual binding system. AB - Plasmodium sporozoites, the transmission form of the malaria parasite, successively invade salivary glands in the mosquito vector and the liver in the mammalian host. Sporozoite capacity to invade host cells is mechanistically related to their ability to glide on solid substrates, both activities depending on the transmembrane protein TRAP. Here, we show that loss-of- function mutations in two adhesive modules of the TRAP ectodomain, an integrin-like A-domain and a thrombospondin type I repeat, specifically decrease sporozoite invasion of host cells but do not affect sporozoite gliding and adhesion to cells. Irrespective of the target cell, i.e. in mosquitoes, rodents and cultured human or hamster cells, sporozoites bearing mutations in one module are less invasive, while those bearing mutations in both modules are non-invasive. In Chinese hamster ovary cells, the TRAP modules interact with distinct cell receptors during sporozoite invasion, and thus act as independently active pass keys. As these modules are also present in other members of the TRAP family of proteins in Apicomplexa, they may account for the capacity of these parasites to enter many cell types of phylogenetically distant origins. PMID- 11927545 TI - IdeS, a novel streptococcal cysteine proteinase with unique specificity for immunoglobulin G. AB - Recent work from several laboratories has demonstrated that proteolytic mechanisms significantly contribute to the molecular interplay between Streptococcus pyogenes, an important human pathogen, and its host. Here we describe the identification, purification and characterization of a novel extracellular cysteine proteinase produced by S.pyogenes. This enzyme, designated IdeS for Immunoglobulin G-degrading enzyme of S.pyogenes, is distinct from the well-characterized streptococcal cysteine proteinase, SpeB, and cleaves human IgG in the hinge region with a high degree of specificity. Thus, other human proteins, including immunoglobulins M, A, D and E, are not degraded by IdeS. The enzyme efficiently cleaves IgG antibodies bound to streptococcal surface structures, thereby inhibiting the killing of S.pyogenes by phagocytic cells. This and additional observations on the distribution and expression of the ideS gene indicate that IdeS represents a novel and significant bacterial virulence determinant, and a potential therapeutic target. PMID- 11927546 TI - Mitochondria are morphologically and functionally heterogeneous within cells. AB - We investigated whether mitochondria represent morphologically continuous and functionally homogenous entities within single intact cells. Physical continuity of mitochondria was determined by three-dimensional reconstruction of fluorescence from mitochondrially targeted DsRed1 or calcein. The mitochondria of HeLa, PAEC, COS-7, HUVEC, hepatocytes, cortical astrocytes and neuronal cells all displayed heterogeneous distributions and were of varying sizes. There was a denser aggregation of mitochondria in perinuclear positions than in the cell periphery, where individual isolated mitochondria could be seen clearly. Using fluorescence-recovery after photobleaching, we observed that DsRed1 and calcein were highly mobile within the matrix of individual mitochondria, and that mitochondria within a cell were not lumenally continuous. Mitochondria were not electrically coupled, since only individual mitochondria were observed to depolarize following irradiation of TMRE-loaded cells. Functional heterogeneity of mitochondria in single cells was observed with respect to membrane potential, sequestration of hormonally evoked cytosolic calcium signals and timing of permeability transition pore opening in response to tert-butyl hydroperoxide. Our data indicate that mitochondria within individual cells are morphologically heterogeneous and unconnected, allowing them to have distinct functional properties. PMID- 11927547 TI - Ligands act as pharmacological chaperones and increase the efficiency of delta opioid receptor maturation. AB - The endoplasmic reticulum (ER) is recognized as an important site for regulating cell surface expression of membrane proteins. We recently reported that only a fraction of newly synthesized delta opioid receptors could leave the ER and reach the cell surface, the rest being degraded by proteasomes. Here, we demonstrate that membrane-permeable opioid ligands facilitate maturation and ER export of the receptor, thus acting as pharmacological chaperones. We propose that these ligands stabilize the newly synthesized receptor in the native or intermediate state of its folding pathway, possibly by inducing stabilizing conformational constrains within the hydrophobic core of the protein. The receptor precursors that are retained in the ER thus represent fully competent folding intermediates that can be targets for pharmacological intervention aimed at regulating receptor expression and cellular responsiveness. The pharmacological chaperone action is independent of the intrinsic signaling efficacy of the ligand, since both agonists and antagonists were found to promote receptor maturation. This novel property of G protein-coupled receptor ligands may have important implications when considering their effects on cellular responsiveness during therapeutic treatments. PMID- 11927548 TI - Multiple endocytic trafficking pathways of MHC class I molecules induced by a Herpesvirus protein. AB - The K3 protein of a human tumor-inducing herpesvirus, Kaposi's sarcoma-associated herpesvirus (KSHV), down-regulates major histocompatibility complex (MHC) class I surface expression by increasing the rate of endocytosis. In this report, we demonstrate that the internalization of MHC class I by the K3 protein is the result of multiple, consecutive trafficking pathways that accelerate the endocytosis of class I molecules, redirect them to the trans-Golgi network (TGN), and target MHC class I to the lysosomal compartment. Remarkably, these actions of K3 are functionally and genetically separable; the N-terminal zinc finger motif and the central sorting motif are involved in triggering internalization of MHC class I molecules and redirecting them to the TGN. Subsequently, the C-terminal diacidic cluster region of K3 is engaged in targeting MHC class I molecules to the lysosomal compartment. These results demonstrate a novel trafficking mechanism of MHC class I molecules induced by KSHV K3, which ensures viral escape from host immune effector recognition. PMID- 11927549 TI - Regulation of intracellular trafficking of human CD1d by association with MHC class II molecules. AB - CD1 family members are antigen-presenting molecules capable of presenting bacterial or synthetic glycolipids to T cells. Here we show that a subset of human CD1d molecules are associated with major histocompatibility complex (MHC) class II molecules, both on the cell surface and in the late endosomal/lysosomal compartments where class II molecules transiently accumulate during transport. The interaction is initiated in the endoplasmic reticulum with class II-invariant chain complexes and appears to be maintained throughout the class II trafficking pathway. A truncated form of CD1d which lacks its cytoplasmic YXXZ internalization motif is transported to late endosomal/lysosomal compartments in the presence of class II molecules. Furthermore, the same CD1d deletion mutant is targeted to lysosomal compartments in HeLa cells expressing class II molecules and invariant chain by transfection. The deletion mutant was also found in lysosomal compartments in HeLa cells expressing only the p33 form of the invariant chain. These data suggest that the intracellular trafficking pathway of CD1d may be altered by class II molecules and invariant chain induced during inflammation. PMID- 11927550 TI - Essential role of endophilin A in synaptic vesicle budding at the Drosophila neuromuscular junction. AB - We characterized Drosophila endophilin A (D-endoA), and generated and analysed D endoA mutants. Like its mammalian homologue, D-endoA exhibits lysophosphatidic acid acyl transferase activity and contains a functional SH3 domain. D-endoA is recruited to the sites of endocytosis, as revealed by immunocytochemistry of the neuromuscular junction (NMJ) of mutant L3 larvae carrying the temperature sensitive allele of dynamin, shibire. D-endoA null mutants show severe defects in motility and die at the early L2 larval stage. Mutants with reduced D-endoA levels exhibit a range of defects of synaptic vesicle endocytosis, as observed at L3 larvae NMJs using FM1-43 uptake and electron microscopy. NMJs with an almost complete loss of synaptic vesicles did not show an accumulation of intermediates of the budding process, whereas NMJs with only slightly reduced levels of synaptic vesicles showed a striking increase in early-stage, but not late-stage, budding intermediates at the plasma membrane. Together with results of previous studies, these observations indicate that endophilin A is essential for synaptic vesicle endocytosis, being required from the onset of budding until fission. PMID- 11927551 TI - Membrane transport in Caenorhabditis elegans: an essential role for VPS34 at the nuclear membrane. AB - Here we present a detailed genetic analysis of let-512/vps34 that encodes the Caenorhabditis elegans homologue of the yeast phosphatidylinositol 3-kinase Vps34p. LET-512/VPS34 has essential functions and is ubiquitously expressed in all tissues and developmental stages. It accumulates at a perinuclear region, and mutations in let-512/vps34 result in an expansion of the outer nuclear membrane as well as in a mislocalization and subsequent complete lack of expression of LRP 1, a C.elegans LDL receptor normally associated with the apical surface of hypodermal cells. Using a GFP::2xFYVE fusion protein we found that the phosphatidylinositol 3-phosphate (PtdIns 3-P) product of LET-512/VPS34 is associated with a multitude of intracellular membranes and vesicles located at the periphery, including endocytic vesicles. We propose that LET-512/VPS34 is required for membrane transport from the outer nuclear membrane towards the cell periphery. Thus, LET-512/VPS34 may regulate the secretory pathway in a much broader range of compartments than was previously suggested for the yeast orthologue. PMID- 11927552 TI - The rasGAP-binding protein, Dok-1, mediates activin signaling via serine/threonine kinase receptors. AB - Activins, members of the transforming growth factor-beta family, are pleiotropic growth and differentiation factors. Activin A induces B-cell apoptosis. To identify the genes responsible for activin-induced apoptosis, we performed retrovirus-mediated gene trap screening in a mouse B-cell line. We identified the rasGAP-binding protein Dok-1 (p62) as an essential molecule that links activin receptors with Smad proteins. In B cells overexpressing Dok-1, activin A-induced apoptotic responses were augmented. The expression of bcl-X(L) was down-regulated by inhibition of the ras/Erk pathway. Activin stimulation triggered association of Dok-1 with Smad3, as well as association of Smad3 with Smad4. Dok-1 also associated with both the type I and type II activin receptors. Dok-1 has been characterized previously as a tyrosine-phosphorylated protein acting downstream of the protein tyrosine kinase pathway: intriguingly, activin signaling did not induce tyrosine phosphorylation of Dok-1. These findings indicate that Dok-1 acts as an adaptor protein that links the activin receptors with the Smads, suggesting a novel function for Dok-1 in activin signaling leading to B-cell apoptosis. PMID- 11927553 TI - Thioredoxin-2 (TRX-2) is an essential gene regulating mitochondria-dependent apoptosis. AB - Thioredoxin-2 (Trx-2) is a mitochondria-specific member of the thioredoxin superfamily. Mitochondria have a crucial role in the signal transduction for apoptosis. To investigate the biological significance of Trx-2, we cloned chicken TRX-2 cDNA and generated clones of the conditional Trx-2-deficient cells using chicken B-cell line, DT40. Here we show that TRX-2 is an essential gene and that Trx-2-deficient cells undergo apoptosis upon repression of the TRX-2 transgene, showing an accumulation of intracellular reactive oxygen species (ROS). Cytochrome c is released from mitochondria, while caspase-9 and caspase-3, but not caspase-8, are activated upon inhibition of the TRX-2 transgene. In addition, Trx-2 and cytochrome c are co-immunoprecipitated in an in vitro assay. These results suggest that mitochondrial Trx-2 is essential for cell viability, playing a crucial role in the scavenging ROS in mitochondria and regulating the mitochondrial apoptosis signaling pathway. PMID- 11927554 TI - Tip60 is targeted to proteasome-mediated degradation by Mdm2 and accumulates after UV irradiation. AB - Acetylation is a prominent post-translational modification of nucleosomal histone N-terminal tails, which regulates chromatin accessibility. Accordingly, histone acetyltransferases (HATs) play major roles in processes such as transcription. Here, we show that the HAT Tip60, which is involved in DNA repair and apoptosis following gamma irradiation, is subjected to proteasome-dependent proteolysis. Furthermore, we provide evidence that Mdm2, the ubiquitin ligase of the p53 tumour suppressor, interacts physically with Tip60 and induces its ubiquitylation and proteasome-dependent degradation. Moreover, a ubiquitin ligase-defective mutant of Mdm2 had no effect on Tip60 stability. Our results indicate that Mdm2 targets both p53 and Tip60, suggesting that these two proteins could be co regulated with respect to protein stability. Consistent with this hypothesis, Tip60 levels increased significantly upon UV irradiation of Jurkat cells. Collectively, our results suggest that degradation of Tip60 could be part of the mechanism leading to cell transformation by Mdm2. PMID- 11927555 TI - The fission yeast NIMA kinase Fin1p is required for spindle function and nuclear envelope integrity. AB - NIMA kinases appear to be the least functionally conserved mitotic regulators, being implicated in chromosome condensation in fungi and in spindle function in metazoans. We demonstrate here that the fission yeast NIMA homologue, Fin1p, can induce profound chromosome condensation in the absence of the condensin and topoisomerase II, indicating that Fin1p-induced condensation differs from mitotic condensation. Fin1p expression is transcriptionally and post-translationally cell cycle-regulated, with Fin1p kinase activity maximal from the metaphase-anaphase transition to G(1). Fin1p is localized to the spindle pole body and fin1Delta cells are hypersensitive to anti-microtubule drugs, synthetically lethal with a number of spindle mutants and require the spindle checkpoint for viability. Moreover, fin1Delta cells show unusual and extensive elaborations of the nuclear envelope. These data support a role for Fin1p in spindle function and nuclear envelope transactions at or after the metaphase-anaphase transition that may be generally applicable to other NIMA-family members. PMID- 11927556 TI - Human Mps1 kinase is required for the spindle assembly checkpoint but not for centrosome duplication. AB - Budding yeast Mps1p kinase has been implicated in both the duplication of microtubule-organizing centers and the spindle assembly checkpoint. Here we show that hMps1, the human homolog of yeast Mps1p, is a cell cycle-regulated kinase with maximal activity during M phase. hMps1 localizes to kinetochores and its activity and phosphorylation state increase upon activation of the mitotic checkpoint. By antibody microinjection and siRNA, we demonstrate that hMps1 is required for human cells to undergo checkpoint arrest in response to microtubule depolymerization. In contrast, centrosome (re-)duplication as well as cell division occur in the absence of hMps1. We conclude that hMps1 is required for the spindle assembly checkpoint but not for centrosome duplication. PMID- 11927557 TI - Casein kinase I phosphorylates the Armadillo protein and induces its degradation in Drosophila. AB - Casein kinase I (CKI) was recently reported as a positive regulator of Wnt signaling in vertebrates and Caenorhabditis elegans. To elucidate the function of Drosophila CKI in the wingless (Wg) pathway, we have disrupted its function by double-stranded RNA-mediated interference (RNAi). While previous findings were mainly based on CKI overexpression, this is the first convincing loss-of-function analysis of CKI. Surprisingly, CKIalpha- or CKIepsilon-RNAi markedly elevated the Armadillo (Arm) protein levels in Drosophila Schneider S2R+ cells, without affecting its mRNA levels. Pulse-chase analysis showed that CKI-RNAi stabilizes Arm protein. Moreover, Drosophila embryos injected with CKIalpha double-stranded RNA showed a naked cuticle phenotype, which is associated with activation of Wg signaling. These results indicate that CKI functions as a negative regulator of Wg/Arm signaling. Overexpression of CKIalpha induced hyper-phosphorylation of both Arm and Dishevelled in S2R+ cells and, conversely, CKIalpha-RNAi reduced the amount of hyper-modified forms. His-tagged Arm was phosphorylated by CKIalpha in vitro on a set of serine and threonine residues that are also phosphorylated by Zeste-white 3. Thus, we propose that CKI phosphorylates Arm and stimulates its degradation. PMID- 11927559 TI - Regulated nuclear import of the STAT1 transcription factor by direct binding of importin-alpha. AB - Signal transducers and activators of transcription (STATs) reside in a latent state in the cytoplasm of the cell, but accumulate in the nucleus in response to cytokines or growth factors. Localization in the nucleus occurs following STAT tyrosine phosphorylation and dimerization. In this report we demonstrate a direct interaction of importin-alpha5 with tyrosine-phosphorylated STAT1 dimers, and provide evidence that a nuclear localization signal (NLS) exists in an inactive state within a STAT1 monomer. A mutation in STAT1 leucine 407 (L407A) is characterized, which generates a protein that is accurately tyrosine phosphorylated in response to interferon, dimerizes and binds DNA, but does not localize to the nucleus. The import defect of STAT1(L407A) appears to be a consequence of the inability of this protein to be recognized by its import shuttling receptor. In addition, we demonstrate that STAT1 binding to specific target DNA effectively blocks importin-alpha5 binding. This result may play a role in localizing STAT1 to its destination in the nucleus, and in releasing importin-alpha5 from STAT1 for recycling back to the cytoplasm. PMID- 11927558 TI - Balancing the activation state of the endothelium via two distinct TGF-beta type I receptors. AB - The generation of mice lacking specific components of the transforming growth factor-beta (TGF-beta) signal tranduction pathway shows that TGF-beta is a key player in the development and physiology of the cardiovascular system. Both pro- and anti-angiogenic properties have been ascribed to TGF-beta, for which the molecular mechanisms are unclear. Here we report that TGF-beta can activate two distinct type I receptor/Smad signalling pathways with opposite effects. TGF-beta induces phosphorylation of Smad1/5 and Smad2 in endothelial cells and these effects can be blocked upon selective inhibition of ALK1 or ALK5 expression, respectively. Whereas the TGF-beta/ALK5 pathway leads to inhibition of cell migration and proliferation, the TGF-beta/ALK1 pathway induces endothelial cell migration and proliferation. We identified genes that are induced specifically by TGF-beta-mediated ALK1 or ALK5 activation. Id1 was found to mediate the TGF beta/ALK1-induced (and Smad-dependent) migration, while induction of plasminogen activator inhibitor-1 by activated ALK5 may contribute to the TGF-beta-induced maturation of blood vessels. Our results suggest that TGF-beta regulates the activation state of the endothelium via a fine balance between ALK5 and ALK1 signalling. PMID- 11927560 TI - The Paf1 complex physically and functionally associates with transcription elongation factors in vivo. AB - We are using biochemical and genetic approaches to study Rtf1 and the Spt4-Spt5 complex, which independently have been implicated in transcription elongation by RNA polymerase II. Here, we report a remarkable convergence of these studies. First, we purified Rtf1 and its associated yeast proteins. Combining this approach with genetic analysis, we show that Rtf1 and Leo1, a protein of unknown function, are members of the RNA polymerase II-associated Paf1 complex. Further analysis revealed allele-specific genetic interactions between Paf1 complex members, Spt4-Spt5, and Spt16-Pob3, the yeast counterpart of the human elongation factor FACT. In addition, we independently isolated paf1 and leo1 mutations in an unbiased genetic screen for suppressors of a cold-sensitive spt5 mutation. These genetic interactions are supported by physical interactions between the Paf1 complex, Spt4-Spt5 and Spt16-Pob3. Finally, we found that defects in the Paf1 complex cause sensitivity to 6-azauracil and diminished PUR5 induction, properties frequently associated with impaired transcription elongation. Taken together, these data suggest that the Paf1 complex functions during the elongation phase of transcription in conjunction with Spt4-Spt5 and Spt16-Pob3. PMID- 11927562 TI - Impaired postnatal hepatocyte proliferation and liver regeneration in mice lacking c-jun in the liver. AB - Mice lacking the AP-1 transcription factor c-jun die at mid-gestation showing heart defects and impaired hepatogenesis. To inactivate c-jun in hepatocytes, mice carrying a floxed c-jun allele were generated. Perinatal liver-specific c jun deletion caused reduced hepatocyte proliferation and decreased body size. After partial hepatectomy, half of the mutants died and liver regeneration was impaired. This phenotype was not present in mice lacking the N-terminal phosphorylation sites of c-Jun. The failure to regenerate was accompanied by increased cell death and lipid accumulation in hepatocytes. Moreover, cyclin dependent kinases and several cell cycle regulators were affected, resulting in inefficient G(1)-S phase progression. These studies identify c-Jun as a critical regulator of hepatocyte proliferation and survival during liver development and regeneration. PMID- 11927561 TI - GAGA can mediate enhancer function in trans by linking two separate DNA molecules. AB - Enhancers have been defined as cis-acting DNA sequences that stimulate transcription from a linked promoter in a distance- and orientation-independent manner. How enhancers activate gene transcription over vast chromosomal distances within metazoan genomes remains poorly understood. Here, we show that the transcription factor GAGA can stimulate transcription by linking an enhancer to its cognate promoter. Strikingly, in addition to facilitating activation by a remote enhancer in cis, GAGA can direct activation of a promoter by an enhancer located on a separate DNA molecule. Enhancer function in trans is critically dependent on POZ domain-mediated GAGA oligomerization, enabling GAGA to bind two DNA molecules simultaneously. Transcriptional activation by an enhancer functioning in trans was observed both in transfected cells and in reconstituted transcription reactions. We propose that GAGA facilitates long-range activation by providing a protein bridge that mediates enhancer-promoter communication. PMID- 11927563 TI - TbMP81 is required for RNA editing in Trypanosoma brucei. AB - Most mitochondrial mRNAs are edited in Trypano soma brucei by a series of steps that are catalyzed by a multienzyme complex that is in its initial stages of characterization. RNA interference (RNAi)-mediated repression of the expression of TbMP81, a zinc finger protein component of the complex, inhibited growth of bloodstream and insect forms, and blocked in vivo RNA editing. This repression preferentially inhibited insertion editing compared with deletion editing in vitro. It resulted in reduced specific endoribonucleolytic cleavage and a greater reduction of U addition and associated RNA ligation activities than U removal and associated RNA ligation activities. The repressed cells retained 20S editing complexes with several demonstrable proteins and adenylatable TbMP52 RNA ligase, but adenlyatable TbMP48 was not detected. Elimination of TbMP48 by RNAi repression did not inhibit cell growth or in vivo editing in either bloodstream or procyclic forms. These results indicate that TbMP81 is required for RNA editing and suggest that the editing complex is functionally partitioned. PMID- 11927564 TI - Dual requirement for yeast hnRNP Nab2p in mRNA poly(A) tail length control and nuclear export. AB - Recent studies of mRNA export factors have provided additional evidence for a mechanistic link between mRNA 3'-end formation and nuclear export. Here, we identify Nab2p as a nuclear poly(A)-binding protein required for both poly(A) tail length control and nuclear export of mRNA. Loss of NAB2 expression leads to hyperadenylation and nuclear accumulation of poly(A)(+) RNA but, in contrast to mRNA export mutants, these defects can be uncoupled in a nab2 mutant strain. Previous studies have implicated the cytoplasmic poly(A) tail-binding protein Pab1p in poly(A) tail length control during polyadenylation. Although cells are viable in the absence of NAB2 expression when PAB1 is overexpressed, Pab1p fails to resolve the nab2Delta hyperadenylation defect even when Pab1p is tagged with a nuclear localization sequence and targeted to the nucleus. These results indicate that Nab2p is essential for poly(A) tail length control in vivo, and we demonstrate that Nab2p activates polyadenylation, while inhibiting hyperadenylation, in the absence of Pab1p in vitro. We propose that Nab2p provides an important link between the termination of mRNA polyadenylation and nuclear export. PMID- 11927565 TI - Trm7p catalyses the formation of two 2'-O-methylriboses in yeast tRNA anticodon loop. AB - The genome of Saccharomyces cerevisiae encodes three close homologues of the Escherichia coli 2'-O-rRNA methyltransferase FtsJ/RrmJ, designated Trm7p, Spb1p and Mrm2p. We present evidence that Trm7p methylates the 2'-O-ribose of nucleotides at positions 32 and 34 of the tRNA anticodon loop, both in vivo and in vitro. In a trm7Delta strain, which is viable but grows slowly, translation is impaired, thus indicating that these tRNA modifications could be important for translation efficiency. We discuss the emergence of a family of three 2'-O-RNA methyltransferases in Eukaryota and one in Prokaryota from a common ancestor. We propose that each eukaryotic enzyme is located in a different cell compartment, in which it would methylate a different RNA that can adopt a very similar secondary structure. PMID- 11927566 TI - The large subunit of initiation factor aIF2 is a close structural homologue of elongation factors. AB - The heterotrimeric factor e/aIF2 plays a central role in eukaryotic/archaeal initiation of translation. By delivering the initiator methionyl-tRNA to the ribosome, e/aIF2 ensures specificity of initiation codon selection. The three subunits of aIF2 from the hyperthermophilic archaeon Pyrococcus abyssi could be overproduced in Escherichia coli. The beta and gamma subunits each contain a tightly bound zinc. The large gamma subunit is shown to form the structural core for trimer assembly. The crystal structures of aIF2gamma, free or complexed to GDP-Mg(2+) or GDPNP-Mg(2+), were resolved at resolutions better than 2 A. aIF2gamma displays marked similarities to elongation factors. A distinctive feature of e/aIF2gamma is a subdomain containing a zinc-binding knuckle. Examination of the nucleotide-complexed aIF2gamma structures suggests mechanisms of action and tRNA binding properties similar to those of an elongation factor. Implications for the mechanism of translation initiation in both eukarya and archaea are discussed. In particular, positioning of the initiator tRNA in the ribosomal A site during the search for the initiation codon is envisaged. PMID- 11927567 TI - Differential mRNA translation and meiotic progression require Cdc2-mediated CPEB destruction. AB - Translational activation of several dormant mRNAs in vertebrate oocytes is mediated by cytoplasmic polyadenylation, a process controlled by the cytoplasmic polyadenylation element (CPE) and its binding protein CPEB. The translation of CPE-containing mRNAs does not occur en masse at any one time, but instead is temporally regulated. We show here that in Xenopus, partial destruction of CPEB controls the temporal translation of CPE-containing mRNAs. While some mRNAs, such as the one encoding Mos, are polyadenylated at prophase I, the polyadenylation of cyclin B1 mRNA requires the partial destruction of CPEB that occurs at metaphase I. CPEB destruction is mediated by a PEST box and Cdc2-catalyzed phosphorylation, and is essential for meiotic progression to metaphase II. CPEB destruction is also necessary for mitosis in the early embryo. These data indicate that a change in the CPEB:CPE ratio is necessary to activate mRNAs at metaphase I and drive the cells' entry into metaphase II. PMID- 11927568 TI - Structure of small protein B: the protein component of the tmRNA-SmpB system for ribosome rescue. AB - Small protein B (SmpB) is an essential component of the highly conserved tmRNA SmpB system that has the dual function of releasing stalled ribosomes from damaged messenger RNAs and targeting incompletely synthesized protein fragments for degradation. Nuclear magnetic resonance (NMR) analysis of SmpB from Aquifex aeolicus revealed an antiparallel beta-barrel structure, with three helices packed outside the core of the barrel. While the overall structure of SmpB appears to be unique, the structure does contain an embedded oligonucleotide binding fold; in this respect SmpB has similarity to several other RNA-binding proteins that are known to be associated with translation, including IF1, ribosomal protein S17 and the N-terminal domain of aspartyl tRNA synthetase. Conserved amino acids on the protein surface that are most likely to directly interact with the tmRNA were identified. The presence of widely separated clusters of conserved amino acids suggests that SmpB could function either by stabilizing two distal regions of the tmRNA, or by facilitating an interaction between the tmRNA and another component of the translational apparatus. PMID- 11927569 TI - Structure of the RPA trimerization core and its role in the multistep DNA-binding mechanism of RPA. AB - The human single-stranded DNA-binding protein, replication protein A (RPA) binds DNA in at least two different modes: initial [8-10 nucleotides (nt)] and stable ( approximately 30 nt). Switching from 8 to 30 nt mode is associated with a large conformational change. Here we report the 2.8 A structure of the RPA trimerization core comprising the C-terminal DNA-binding domain of subunit RPA70 (DBD-C), the central DNA-binding domain of subunit RPA32 (DBD-D) and the entire RPA14 subunit. All three domains are built around a central oligonucleotide/oligosaccharide binding (OB)-fold and flanked by a helix at the C terminus. Trimerization is mediated by three C-terminal helices arranged in parallel. The OB-fold of DBD-C possesses unique structural features; embedded zinc ribbon and helix-turn-helix motifs. Using time-resolved proteolysis with trypsin, we demonstrate that the trimerization core does not contribute to the binding with substrates of 10 nt, but interacts with oligonucleotides of 24 nt. Taken together, our data indicate that switching from 8-10 to 30 nt mode is mediated by DNA binding with the trimerization core. PMID- 11927570 TI - Compatible bacterial plasmids are targeted to independent cellular locations in Escherichia coli. AB - Targeting of DNA molecules to specific subcellular positions is essential for efficient segregation, but the mechanisms underlying these processes are poorly understood. In Escherichia coli, several plasmids belonging to different incompatibility groups (F, P1 and RK2) localize preferentially near the midcell and quartercell positions. Here we compare the relative positions of these three plasmids using fluorescence in situ hybridization. When plasmids F and P1 were localized simultaneously using differentially labeled probes, the majority of foci (approximately 75%) were well separated from each other. Similar results were found when we compared the subcellular localization of F with RK2, and RK2 with P1: regardless of the number of foci per cell or growth conditions, most of the foci (70-80%) were not in close proximity to one another. We also localized RK2 in Pseudomonas aeruginosa and Vibrio cholerae, and found that plasmid RK2 localization is conserved across bacterial species. Our results suggest that each plasmid has its own unique subcellular address, implying a mechanism for the stable co-existence of plasmids in which subcellular targeting plays a major role. PMID- 11927571 TI - A few amino acid substitutions can convert deoxyribonucleoside kinase specificity from pyrimidines to purines. AB - In mammals, the four native deoxyribonucleosides are phosphorylated to the corresponding monophosphates by four deoxyribonucleoside kinases, which have specialized substrate specificities. These four enzymes are likely to originate from a common progenitor kinase. Insects appear to have only one multisubstrate deoxyribonucleoside kinase (dNK, EC 2.7.1.145), which prefers pyrimidine nucleosides, but can also phosphorylate purine substrates. When the structures of the human deoxyguanosine kinase (dGK, EC 2.7.1.113) and the dNK from Drosophila melanogaster were compared, a limited number of amino acid residues were identified and proposed to be responsible for the substrate specificity. Three of these key residues in Drosophila dNK were then mutagenized and the mutant enzymes were characterized regarding their ability to phosphorylate native deoxyribonucleosides and nucleoside analogs. The mutations converted the dNK substrate specificity from predominantly pyrimidine specific into purine specific. A similar scenario could have been followed during the evolution of kinases. Upon gene duplication of the progenitor kinase, only a limited number of single amino acid changes has taken place in each copy and resulted in substrate specialized enzymes. PMID- 11927572 TI - Carboxyl-terminal peptide of beta-amyloid precursor protein blocks inositol 1,4,5 trisphosphate-sensitive Ca2+ release in Xenopus laevis oocytes. AB - The effects of Alzheimer's disease-related amyloidogenic peptides on inositol 1,4,5-trisphosphate receptor-mediated Ca(2+) mobilization were examined in Xenopus laevis oocytes. Intracellular Ca(2+) was monitored by electrophysiological measurement of the endogenous Ca(2+)-activated Cl(-) current. Application of a hyperpolarizing pulse released intracellular Ca(2+) in oocytes primed by pre-injection of a non-metabolizable inositol 1,4,5 trisphosphate analogue. The carboxyl terminus of the amyloid precursor protein inhibited inositol 1,4,5-trisphosphate receptor-mediated intracellular Ca(2+) release in a dose-dependent manner. Equimolar beta-amyloid peptides Abeta(1-40) or Abeta(1-42) had no effect, and whereas a truncated carboxyl terminus lacking the Abeta domain was equipotent to the full-length one, a carboxyl terminus fragment lacking the NPTY sequence was less effective than the full-length fragment. The inhibition induced by the carboxyl terminus was not associated with the block of the Ca(2+)-dependent Cl(-) channel itself or compromised Ca(2+) influx. We conclude that the carboxyl terminus of the amyloid precursor protein inhibits inositol 1,4,5-trisphosphate-sensitive Ca(2+) release and could thus disrupt Ca(2+) homeostasis and that the carboxyl terminus is much more effective than the beta-amyloid fragments used. By perturbing the coupling of inositol 1,4,5-trisphosphate and Ca(2+) release, the carboxyl terminus of the amyloid precursor protein can potentially be involved in inducing the neural toxicity characteristic of Alzheimer's disease. PMID- 11927573 TI - Structural and functional analysis of the human mitotic-specific ubiquitin conjugating enzyme, UbcH10. AB - Cell cycle progression is controlled at several different junctures by the targeted destruction of cell cycle regulatory proteins. These carefully orchestrated events include the destruction of the securin protein to permit entry into anaphase, and the destruction of cyclin B to permit exit from mitosis. These destruction events are mediated by the ubiquitin/proteasome system. The human ubiquitin-conjugating enzyme, UbcH10, is an essential mediator of the mitotic destruction events. We report here the 1.95-A crystal structure of a mutant UbcH10, in which the active site cysteine has been replaced with a serine. Functional analysis indicates that the mutant is active in accepting ubiquitin, although not as efficiently as wild-type. Examination of the crystal structure reveals that the NH2-terminal extension in UbcH10 is disordered and that a conserved 3(10)-helix places a lysine residue near the active site. Analysis of relevant mutants demonstrates that for ubiquitin-adduct formation the presence or absence of the NH2-terminal extension has little effect, whereas the lysine residue near the active site has significant effect. The structure provides additional insight into UbcH10 function including possible sites of interaction with the anaphase promoting complex/cyclosome and the disposition of a putative destruction box motif in the structure. PMID- 11927574 TI - Probing interactions between the U2 small nuclear ribonucleoprotein and the DEAD box protein, Prp5. AB - Pre-mRNA binding to the yeast U2 small nuclear ribonucleoprotein (snRNP) during prespliceosome formation requires ATP hydrolysis, the highly conserved UACUAAC box of the branch point region of the pre-mRNA, and several factors. Here we analyzed the binding of a radiolabeled 2'-O-methyl oligonucleotide complementary to U2 small nuclear RNA to study interactions between the UACUAAC box, U2 snRNP, and Prp5p, a DEAD box protein necessary for prespliceosome formation. Binding of the 2'-O-methyl oligonucleotide to the U2 snRNP in yeast cell extract was assayed by gel electrophoresis. Binding was rapid, enhanced by ATP, and dependent on the integrity and conformation of the U2 snRNP. It was also stimulated by Prp5p that was found to associate physically with U2 snRNP. In vitro heat inactivation of the temperature-sensitive prp5-1 mutant extract decreased oligonucleotide binding to U2 and the ATP enhancement of binding by 3-fold. Furthermore, the temperature sensitive prp5-1 mutation maps to the ATP-binding motif I within the helicase like domain. Thus the catalytic activity of Prp5p likely promotes a conformational change in the U2 snRNP. PMID- 11927576 TI - Positive contribution of hydration structure on the surface of human lysozyme to the conformational stability. AB - Water molecules make a hydration structure with the network of hydrogen bonds, covering on the surface of proteins. To quantitatively estimate the contribution of the hydration structure to protein stability, a series of hydrophilic mutant human lysozymes (Val to Ser, Tyr, Asp, Asn, and Arg) modified at three different positions on the surface, which are located in the alpha-helix (Val-110), the beta-sheet (Val-2), and the loop (Val-74), were constructed. Their thermodynamic parameters of denaturation and crystal structures were examined by calorimetry and by x-ray crystallography at 100 K, respectively. The introduced polar residues made hydrogen bonds with protein atoms and/or water molecules, sometimes changing the hydration structure around the mutation site. Changes in the stability of the mutant proteins can be evaluated by a unique equation that considers the conformational changes resulting from the substitutions. Using this analysis, the relationship between the changes in the stabilities and the hydration structures for mutant human lysozymes substituted on the surface could be quantitatively estimated. The analysis indicated that the hydration structure on protein surface plays an important role in determining the conformational stability of the protein. PMID- 11927575 TI - Cellular responses to the DNA strand-scission enediyne C-1027 can be independent of ATM, ATR, and DNA-PK kinases. AB - The current paradigm based upon ionizing radiation (IR) studies states that cells deficient in either ataxia-telangiectasia-mutated kinase (ATM) or related phosphatidylinositol 3 (PI 3) -kinases (ATR and DNA-PK) are hypersensitive to DNA strand breaks because they are unable to rapidly activate downstream effectors such as p53. Here we have contrasted cell responses to IR and C-1027, a radiomimetic antibiotic that induces DNA strand breaks. At equal levels of DNA double strand breaks, cell lines with inactive ATM or other phosphatidylinositol 3-kinases displayed classical hypersensitivity to IR but not to C-1027. Moreover, phosphorylation of p53 Ser-15 induced by C-1027 was independent of ATM, ATR, or DNA-PK function. We have concluded that the model based on IR studies cannot always be directly applied to DNA damage induced by other strand-scission agents. PMID- 11927577 TI - Biochemical characterization of the kinase domain of the rice disease resistance receptor-like kinase XA21. AB - The rice disease resistance gene, Xa21, encodes a receptor kinase-like protein consisting of leucine-rich repeats in the putative extracellular domain and a serine/threonine kinase in the putative intracellular domain. The putative XA21 kinase domain was expressed as maltose-binding and glutathione S-transferase fusion proteins in Escherichia coli. The fusion proteins are capable of autophosphorylation. Phosphoamino acid analysis of the glutathione S-transferase fusion protein indicates that only serine and threonine residues are phosphorylated. The relative phosphorylation rate of the XA21 kinase against increasing enzyme concentrations follows a first-order rather than second-order kinetics, indicating an intramolecular phosphorylation mechanism. Moreover, the active XA21 kinase cannot phosphorylate a kinase-deficient mutant of XA21 kinase. The enzymatic activity of the XA21 kinase in a buffer containing Mn(2+) is at least 15 times higher than that with Mg(2+). The K(m) and V(max) of XA21 kinase for ATP are 0.3 microm and 8.4 nmol/mg/min, respectively. Tryptic phosphopeptide mapping reveals that multiple sites on the XA21 kinase are phosphorylated. Finally, our data suggest that the region of XA21 kinase corresponding to the RD kinase activation domain is not phosphorylated, revealing a distinct mode of action compared with the tomato Pto serine/threonine kinase conferring disease resistance. PMID- 11927578 TI - Ca2+-dependent translocation of the calcyclin-binding protein in neurons and neuroblastoma NB-2a cells. AB - The calcyclin-binding protein (CacyBP) binds calcyclin (S100A6) at physiological levels of [Ca(2+)] and is highly expressed in brain neurons. Subcellular localization of CacyBP was examined in neurons and neuroblastoma NB-2a cells at different [Ca(2+)](i). Immunostaining indicates that CacyBP is present in the cytoplasm of unstimulated cultured neurons in which resting [Ca(2+)](i) is known to be approximately 50 nm. When [Ca(2+)](i) was increased to above 300 nm by KCl treatment, the immunostaining was mainly apparent as a ring around the nucleus. Such perinuclear localization of CacyBP was observed in untreated neuroblastoma NB-2a cells in which [Ca(2+)](i) is approximately 120 nm. An additional increase in [Ca(2+)](i) to above 300 nm by thapsigargin treatment did not change CacyBP localization. However, when [Ca(2+)](i) in NB-2a cells dropped to 70 nm, because of BAPTA/AM treatment, perinuclear localization was diminished. Ca(2+)-induced translocation of CacyBP was confirmed by immunogold electron microscopy and by fluorescence of NB-2a cells transfected with an EGFP-CacyBP vector. Recombinant CacyBP can be phosphorylated by protein kinase C in vitro. In untreated neuroblastoma NB-2a cells, CacyBP is phosphorylated on a serine residue(s), but exists in the dephosphorylated form in BAPTA/AM-treated cells. Thus, phosphorylation of CacyBP occurs in the same [Ca(2+)](i) range that leads to its perinuclear translocation. PMID- 11927579 TI - Infection of glioma cells with Sindbis virus induces selective activation and tyrosine phosphorylation of protein kinase C delta. Implications for Sindbis virus-induced apoptosis. AB - Sindbis virus (SV) is an alpha virus used as a model for studying the role of apoptosis in virus infection. In this study, we examined the role of protein kinase C (PKC) in the apoptosis induced by SVNI, a virulent strain of SV. Infection of C6 cells with SVNI induced a selective translocation of PKCdelta to the endoplasmic reticulum and its tyrosine phosphorylation. The specific PKCdelta inhibitor rottlerin and a PKCdelta kinase-dead mutant increased the apoptosis induced by SVNI. To examine the role of the tyrosine phosphorylation of PKCdelta in the apoptosis induced by SVNI we used a PKCdelta mutant in which five tyrosine residues were mutated to phenylalanine (PKCdelta5). PKCdelta5-overexpressing cells exhibited increased apoptosis in response to SVNI as compared with control cells and to cells overexpressing PKCdelta. SVNI also increased the cleavage of caspase 3 in cells overexpressing PKCdelta5 but did not induce cleavage of PKCdelta or PKCdelta5. Using single tyrosine mutants, we identified tyrosines 52, 64, and 155 as the phosphorylation sites associated with the apoptosis induced by SVNI. We conclude that PKCdelta exerts an inhibitory effect on the apoptosis induced by SV and that phosphorylation of PKCdelta on specific tyrosines is required for this function. PMID- 11927580 TI - Assembly of the bacteriophage T4 helicase: architecture and stoichiometry of the gp41-gp59 complex. AB - The bacteriophage T4 59 protein (gp59) plays an essential role in recombination and replication by mediating the assembly of the gene 41 helicase (gp41) onto DNA. gp59 is required to displace the gp32 single-stranded binding protein on the lagging strand to expose a site for helicase binding. To gain a better understanding of the mechanism of helicase assembly, the architecture and stoichiometry of the gp41-gp59 complex were investigated. Both the N and C termini of gp41 were found to lie close to or in the gp41-gp41 subunit interface and interact with gp59. The site of interaction of gp41 on gp59 is proximal to Cys-215 of gp59. Binding of gp41 to gp59 stimulates a conformational change in the protein resulting in hexamer formation of gp59, and gp59 likewise stimulates oligomer formation of gp41. The gp59 subunits in this complex are arranged in a head to head orientation, such that Cys-42 of one subunit is in close proximity to Cys-42 on an adjacent subunit, and Cys-215 on one subunit is close to Cys-215 on a neighboring subunit. As the helicase is loaded onto DNA, a conformational change in the gp41-gp59 complex occurs, which may serve to displace gp32 from the lagging strand and load the hexameric helicase in its place. PMID- 11927582 TI - Mechanistic role of residue Gln151 in error prone DNA synthesis by human immunodeficiency virus type 1 (HIV-1) reverse transcriptase (RT). Pre-steady state kinetic study of the Q151N HIV-1 RT mutant with increased fidelity. AB - It has previously been reported that mutations in the Gln(151) residue of human immunodeficiency virus type 1 (HIV-1) reverse transcriptase (RT) greatly enhance RT fidelity. In this study, we employed pre-steady state kinetic assays to elucidate the mechanistic role of residue Gln(151) in highly error prone DNA synthesis by HIV-1 RT. Using our Q151N high fidelity mutant, which is structurally altered in its ability to interact with the 3'-OH on the sugar moiety of the incoming deoxynucleotide triphosphate (dNTP), we examined how this change in RT-dNTP interaction affects HIV-1 RT fidelity. First, we found the binding affinity (K(D)) of wild type and Q151N RT proteins to different template/primers to be similar. These results indicate that the Gln(151) residue is not involved in the formation of the binary complex (RT.template/primer) during DNA polymerization. We also found that by changing residue 151 from a Gln- >Asn, the maximum rate of dNTP incorporation (k(pol)) for both correct and incorrect dNTPs was not affected. In contrast, the ability of the Q151N mutant to bind both correct and incorrect dNTPs (K(d)) was diminished. The Q151N mutant was 120-fold less efficient at binding correct dNTP than wild type RT, and its decrease in binding was such that we were unable to measure the actual binding affinity of Q151N for incorrect dNTPs. Presumably, the fidelity increase observed during the steady state is explained by this defect in Q151N binding to incorrect dNTP. In wild type RT, residue Gln(151) is important for tight binding of incorrect dNTPs and may contribute to the low fidelity nature of HIV-1 RT. Since the Q151N mutation also alters RT binding to correct dNTPs, the wild type Gln(151) residue may play an important role in efficient binding of RT to correct dNTPs. Our findings suggest that residue Gln(151) is an important element for the execution of both highly error prone and efficient DNA synthesis by HIV-1 RT. PMID- 11927581 TI - Binding of hydrophobic peptides to several non-catalytic sites promotes peptide hydrolysis by all active sites of 20 S proteasomes. Evidence for peptide-induced channel opening in the alpha-rings. AB - The eukaryotic 20 S proteasome contains the following 6 active sites: 2 chymotrypsin-like, 2 trypsin-like, and 2 caspase-like. We previously showed that hydrophobic peptide substrates of the chymotrypsin-like sites allosterically stimulate peptide hydrolysis by the caspase-like sites and their own cleavage. More thorough analysis revealed that these peptides also stimulate peptide hydrolysis by the trypsin-like site. This general activation by hydrophobic peptides occurred even if the chymotrypsin-like sites were occupied by a covalent inhibitor and was highly cooperative, with an average Hill coefficient of 7. Therefore, this stimulation of peptide hydrolysis at all active sites occurs upon binding of hydrophobic peptides to several non-catalytic sites. The stimulation by hydrophobic peptides was not observed in the yeast Delta N alpha 3 mutant 20 S proteasomes, in 20 S-PA26 complexes, or SDS-activated proteasomes and was significantly lower in 26 S proteasomes, all of which appear to have the gated channel in the alpha-rings in an open conformation and hydrolyze peptides at much faster rates than 20 S proteasomes. Also the hydrophobic peptides altered K(m), V(max) of active sites in a similar fashion as PA26 and the Delta N alpha 3 mutation. The activation by hydrophobic peptides was decreased in K(+)-containing buffer, which favors the closed state of the channels. Therefore, hydrophobic peptides stimulate peptide hydrolysis most likely by promoting the opening of the channels in the alpha-rings. During protein breakdown, this peptide-induced channel opening may function to facilitate the release of products from the proteasome. PMID- 11927583 TI - Efficient rejoining of radiation-induced DNA double-strand breaks in centromeric DNA of human cells. AB - Although major efforts in elucidating different DNA double-strand break (DSB) repair pathways and their contribution to accurate repair or misrepair have been made, little is known about the influence of chromatin structure on the fidelity of DSB repair. Here, the repair of ionizing radiation-induced DSBs was investigated in heterochromatic centromeric regions of human cells in comparison with other genomic locations. A hybridization assay was applied that allows the quantification of correct DSB rejoining events in specific genomic regions by measuring reconstitution of large restriction fragments. We show for two primary fibroblast lines (MRC-5 and 180BR) and an epithelial tumor cell line that restriction fragment reconstitution is considerably more efficient in the centromere than in average genomic locations. Importantly, however, DNA ligase IV deficient 180BR cells show, compared with repair-proficient MRC-5 cells, impaired restriction fragment reconstitution both in average DNA and in the centromere. Thus, the efficient repair of DSBs in centromeric DNA is dependent on functional non-homologous end joining. It is proposed that the condensed chromatin state in the centromere limits the mobility of break ends and leads to enhanced restriction fragment reconstitution by increasing the probability for rejoining correct break ends. PMID- 11927584 TI - Human neuropathy target esterase catalyzes hydrolysis of membrane lipids. AB - A neuronal membrane protein, neuropathy target esterase (NTE), reacts with those organophosphates that initiate a syndrome of axonal degeneration. NTE has homologues in Drosophila and yeast and is detected in vitro by assays with a non physiological ester substrate, phenyl valerate. We report that NEST, the recombinant esterase domain of NTE (residues 727-1216) purified from bacterial lysates, can catalyze hydrolysis of several naturally occurring membrane associated lipids. The active site regions of NEST and calcium-independent phospholipase A(2) (iPLA(2)) share sequence similarity, and the phenyl valerate hydrolase activity of NEST is inhibited by low concentrations of iPLA(2) inhibitors. However, on incubation with NEST, fatty acid was liberated only extremely slowly from the sn-2 position of phospholipids (V(max) approximately 0.01 micromol/min/mg and K(m) approximately 0.4 mm for 1-palmitoyl, 2 oleoylphosphatidylcholine). Comparison of the NEST-mediated generation of (14)C labeled products from two differentially labeled (14)C-phospholipid substrates suggested that a rate-limiting sn-2 cleavage was followed very rapidly by hydrolysis of the resulting lysophospholipid. Among the various naturally occurring lipids tested with NEST, lysophospholipids were by far the most avidly hydrolyzed substrates (V(max) approximately 20 micromol/min/mg and K(m) approximately 0.05 mm for 1-palmitoyl-lysophosphatidylcholine). NEST also catalyzed the hydrolysis of monoacylglycerols, preferring the 1-acyl to the 2 acyl isomer (V(max) approximately 1 micromol/min/mg and K(m) approximately 0.4 mm for 1-palmitoylglycerol). NEST did not catalyze hydrolysis of di- or triacylglycerols or fatty acid amides. This demonstration that membrane lipids are its putative cellular substrates raises the possibility that NTE and its homologues may be involved in intracellular membrane trafficking. PMID- 11927585 TI - Peroxisome proliferator-activated receptor delta activation promotes cell survival following hypertonic stress. AB - COX2-selective non-steroidal anti-inflammatory drugs (NSAIDs) cause selective apoptosis of renal medullary interstitial cells (RMIC) in vivo and reduce their ability to tolerate hypertonic stress in vitro. To determine the mechanism by which COX2 activity promotes RMIC viability, we examined the capacity of COX2 derived prostanoids to promote RMIC survival. Although RMICs synthesize prostaglandin E2 (PGE2) PGI2 > PGF2a > TxA2, only PGI2 enhanced RMIC viability following hypertonic stress. RMICs do not express the prostacyclin receptor, but they do express the prostacyclin responsive nuclear transcription factor peroxisome proliferator-activated receptor delta (PPARdelta). Hypertonic stress increased PGI2 synthesis 330% above base line and also activated a PPARdelta specific reporter (delta response element (DRE)) by 90% above base line. Conversely DRE activity was only inhibited by the COX2-selective inhibitor SC236 but not by a COX1-selective NSAID (SC560). Overexpression of PPARdelta using an adenovirus not only drove DRE activity but also prevented RMIC death due to COX2 inhibition. These studies are consistent with a model whereby hypertonicity activates COX2-derived prostaglandin production, which promotes RMIC viability through PPARdelta. Inhibition of PPARdelta activity may contribute to the renal papillary necrosis associated with analgesic and/or NSAID use. PMID- 11927586 TI - Modeling apoptotic chromatin condensation in normal cell nuclei. Requirement for intranuclear mobility and actin involvement. AB - Hallmarks of the terminal stages of apoptosis are genomic DNA fragmentation and chromatin condensation. Here, we have studied the mechanism of condensation both in vitro and in vivo. We found that DNA fragmentation per se of isolated nuclei from non-apoptotic cells induced chromatin condensation that closely resembles the morphology seen in apoptotic cells, independent of ATP utilization, at physiological ionic strengths. Interestingly, chromatin condensation was accompanied by release of nuclear actin, and both condensation and actin release could be blocked by reversibly pretreating nuclei with Ca2+, Cu2+, diamide, or low pH, procedures shown to stabilize internal nuclear components. Moreover, specific inhibition of nuclear F-actin depolymerization or promotion of its formation also reduced chromatin condensation. Chromatin condensation could also be inhibited by exposing nuclei to reagents that bind to the DNA minor groove, disrupting native nucleosomal DNA wrapping. In addition, in cultured cells undergoing apoptosis, drugs that inhibit depolymerization of actin or bind to the minor groove also reduced chromatin condensation, but not DNA fragmentation. Therefore, the ability of chromatin fragments with intact nucleosomes to form large clumps of condensed chromatin during apoptosis requires the apparent disassembly of internal nuclear structures that may normally constrain chromosome subdomains in non-apoptotic cells. PMID- 11927587 TI - Catalytic mechanism of S-adenosylhomocysteine hydrolase. Site-directed mutagenesis of Asp-130, Lys-185, Asp-189, and Asn-190. AB - S-Adenosylhomocysteine hydrolase (AdoHcyase) catalyzes the hydrolysis of S adenosylhomocysteine to form adenosine and homocysteine. On the bases of crystal structures of the wild type enzyme and the D244E mutated enzyme complexed with 3' keto-adenosine (D244E.Ado*), we have identified the important amino acid residues, Asp-130, Lys-185, Asp-189, and Asn-190, for the catalytic reaction and have proposed a catalytic mechanism (Komoto, J., Huang, Y., Gomi, T., Ogawa, H., Takata, Y., Fujioka, M., and Takusagawa, F. (2000) J. Biol. Chem. 275, 32147 32156). To confirm the proposed catalytic mechanism, we have made the D130N, K185N, D189N, and N190S mutated enzymes and measured the catalytic activities. The catalytic rates (k(cat)) of D130N, K185N, D189N, and N190S mutated enzymes are reduced to 0.7%, 0.5%, 0.1%, and 0.5%, respectively, in comparison with the wild type enzyme, indicating that Asp-130, Lys-185, Asp-189, and Asn-190 are involved in the catalytic reaction. K(m) values of the mutated enzymes are increased significantly, except for the N190S mutation, suggesting that Asp-130, Lys-185, and Asp-189 participate in the substrate binding. To interpret the kinetic data, the oxidation states of the bound NAD molecules of the wild type and mutated enzymes were measured during the catalytic reaction by monitoring the absorbance at 340 nm. The crystal structures of the WT and D244E.Ado*, containing four subunits in the crystallographic asymmetric unit, were re-refined to have the same subunit structures. A detailed catalytic mechanism of AdoHcyase has been revealed based on the oxidation states of the bound NAD and the re-refined crystal structures of WT and D244E.Ado*. Lys-185 and Asp-130 abstract hydrogen atoms from 3'-OH and 4'-CH, respectively. Asp-189 removes a proton from Lys-185 and produces the neutral N zeta (-NH(2)), and Asn-190 facilitates formation of the neutral Lys-185. His-54 and His-300 hold and polarize a water molecule, which nucleophilically attacks the C5'- of 3'-keto-4',5'-dehydroadenosine to produce 3' keto-Ado. PMID- 11927588 TI - Liver receptor homologue-1 (LRH-1) regulates expression of aromatase in preadipocytes. AB - Estrogen biosynthesis from C(19) steroids is catalyzed by aromatase cytochrome P450. Aromatase is expressed in breast adipose tissue through the use of a distal, cytokine-responsive promoter (promoter I.4). Breast tumors, however, secrete soluble factors that stimulate aromatase expression through an alternative proximal promoter, promoter II. In other estrogenic tissues such as ovaries, transcription from promoter II requires the presence of the Ftz-F1 homologue steroidogenic factor-1 (SF-1); adipose tissue, however, does not express SF-1. We have explored the hypothesis that in adipose tissue, an alternative Ftz-F1 family member, liver receptor homologue-1 (LRH-1), substitutes for SF-1 in driving transcription from promoter II. In transient transfection assays using 3T3-L1 preadipocytes, promoter II reporter constructs were modestly (2-3-fold) stimulated by either treatment with activators of protein kinases A or C (PKA/C) or by cotransfection with LRH-1. In combination, these treatments synergistically activated promoter II (>30-fold). Induction by LRH-1 (but not by PKA/C) required an AGGTCA motif at -130 base pairs, to which LRH-1 bound in gel shift assays. Activity of GAL4-LRH-1 fusion proteins was not altered by activators of PKA or PKC. Quantitative real-time PCR revealed that LRH-1 (but not SF-1) is expressed in the preadipocyte fraction of human adipose tissue at levels comparable with that of liver. Differentiation of cultured human preadipocytes into mature adipocytes was associated with a time-dependent induction of peroxisome proliferator-activated receptor-gamma (PPARgamma), and rapid loss of LRH-1 and aromatase expression. We conclude that LRH-1 is a preadipocyte-specific nuclear receptor that regulates expression of aromatase in adipose tissue. Alterations in LRH-1 expression and/or activity in adipose tissue could therefore have considerable effects on local estrogen production and breast cancer development. PMID- 11927589 TI - Role of C-terminal domain and transmembrane helices 5 and 6 in function and quaternary structure of major intrinsic proteins: analysis of aquaporin/glycerol facilitator chimeric proteins. AB - We previously observed that aquaporins and glycerol facilitators exhibit different oligomeric states when studied by sedimentation on density gradients following nondenaturing detergent solubilization. To determine the domains of major intrinsic protein (MIP) family proteins involved in oligomerization, we constructed protein chimeras corresponding to the aquaporin AQPcic substituted in the loop E (including the proximal part of transmembrane domain (TM) 5) and/or the C-terminal part (including the distal part of TM 6) by the equivalent domain of the glycerol channel aquaglyceroporin (GlpF) (chimeras called AGA, AAG, and AGG). The analogous chimeras of GlpF were also constructed (chimeras GAG, GGA, and GAA). cRNA corresponding to all constructs were injected into Xenopus oocytes. AQPcic, GlpF, AAG, AGG, and GAG were targeted to plasma membranes. Water or glycerol membrane permeability measurements demonstrated that only the AAG chimera exhibited a channel function corresponding to water transport. Analysis of all proteins expressed either in oocytes or in yeast by velocity sedimentation on sucrose gradients following solubilization by 2% n-octyl glucoside indicated that only AQPcic and AAG exist in tetrameric forms. GlpF, GAG, and GAA sediment in a monomeric form, whereas GGA and AGG were found mono/dimeric. These data bring new evidence that, within the MIP family, aquaporins and GlpFs behave differently toward nondenaturing detergents. We demonstrate that the C-terminal part of AQPcic, including the distal half of TM 6, can be substituted by the equivalent domain of GlpF (AAG chimera) without modifying the transport specificity. Our results also suggest that interactions of TM 5 of one monomer with TM 1 of the adjacent monomer are crucial for aquaporin tetramer stability. PMID- 11927590 TI - HTLV-1 Tax-associated hTid-1, a human DnaJ protein, is a repressor of Ikappa B kinase beta subunit. AB - hTid-1, a human DnaJ protein, is a novel cellular target for HTLV-1 Tax. Here, we show that hTid-1 represses NF-kappaB activity induced by Tax as well as other activators such as tumor necrosis factor alpha (TNFalpha) and Bcl10. hTid-1 specifically suppresses serine phosphorylation of IkappaBalpha by activated IkappaB kinase beta (IKKbeta), but the activities of other serine kinases including p38, ERK2, and JNK1 are not affected. The suppressive activity of hTid 1 on IKKbeta requires a functional J domain that mediates association with heat shock proteins and results in prolonging the half-life of the NF-kappaB inhibitors IkappaBalpha and IkappaBbeta. Collectively, our data suggest that hTid 1, in association with heat shock proteins, exerts a negative regulatory effect on the NF-kappaB activity induced by various extracellular and intracellular activators including HTLV-1 Tax. PMID- 11927592 TI - A highly effective dominant negative alpha s construct containing mutations that affect distinct functions inhibits multiple Gs-coupled receptor signaling pathways. AB - To investigate the subcellular organization of receptor-G protein signaling pathways, a robust dominant negative alpha(s) mutant containing substitutions that alter distinct functions was produced and tested for its effects on G(s) coupled receptor activity in HEK-293 cells. Mutations in the alpha3beta5 loop region, which increase receptor affinity, decrease receptor-mediated activation, and impair activation of adenylyl cyclase, were combined with G226A, which increases affinity for betagamma, and A366S, which decreases affinity for GDP. This triple alpha(s) mutant can inhibit signaling to G(s) from the luteinizing hormone receptor by 97% and from the calcitonin receptor by 100%. In addition, this alpha(s) mutant blocks all signaling from the calcitonin receptor to G(q). These results lead to two conclusions about receptor-G protein signaling. First, individual receptors have access to multiple types of G proteins in HEK-293 cell membranes. Second, different G protein alpha subunits can compete with each other for binding to the same receptor. This dominant negative alpha(s) construct will be useful for determining interrelationships among distinct receptor-G protein interactions in a wide variety of cells and tissues. PMID- 11927591 TI - Autoubiquitination of the BRCA1*BARD1 RING ubiquitin ligase. AB - The RING finger of BRCA1 confers ubiquitin ligase activity that is markedly enhanced when complexed with another RING-containing protein, BARD1, and is required for the function of this tumor suppressor protein in protecting genomic integrity. Here, we report that co-expression of BRCA1-(1-639) and BARD1 in bacteria can assemble a potent ubiquitin ligase activity. Purified BRCA1-(1 639)*BARD1 stimulated the Ubc5c-mediated monoubiquitination of histone H2A/H2AX in vitro, suggesting a possible role for BRCA1*BARD1 in modifying chromatin structure. Moreover, the truncated BRCA1*BARD1 complex exhibited efficient autoubiquitination activity in vitro capable of assembling non-lysine 48-linked polyubiquitin chains on both BRCA1-(1-639) and BARD1. When co-expressed in cells by transient transfection, the recombinant BRCA1-(1-300).BARD1 complex was found to be associated with polyubiquitin chains, suggesting that BRCA1-(1-300)*BARD1 was ubiquitinated in vivo as well. These results raise the possibility that BRCA1*BARD1 acts to assemble non-lysine 48-linked polyubiquitin chains that may serve as part of a signaling platform required for coordinating DNA repair related events. PMID- 11927593 TI - The newly identified human nuclear protein NXP-2 possesses three distinct domains, the nuclear matrix-binding, RNA-binding, and coiled-coil domains. AB - Using a monoclonal antibody that recognizes a nuclear matrix protein, we selected a cDNA clone from a lambdagt11 human placenta cDNA library. This cDNA encoded a 939-amino acid protein designated nuclear matrix protein NXP-2. Northern blot analysis indicated that NXP-2 was expressed in various tissues at different levels. Forcibly expressed green fluorescent protein-tagged NXP-2 as well as endogenous NXP-2 was localized in the nucleus and distributed to the nuclear matrix. NXP-2 was released from the nuclear matrix when RNase A was included in the buffer for nuclear matrix preparation. Mapping of functional domains was carried out using green fluorescent protein-tagged truncated mutants of NXP-2. The region of amino acids 326-353 was responsible for nuclear matrix binding and contained a cluster of hydrophobic amino acids that was similar to the nuclear matrix targeting signal of acute myeloleukemia protein. The central region (amino acids 500-591) was demonstrated to be required for RNA binding by Northwestern analysis, although NXP-2 lacked a known RNA binding motif. The region of amino acid residues 682-876 was predicted to have a coiled-coil structure. The RNA binding, nuclear matrix-binding, and coiled-coil domains are structurally separated, suggesting that NXP-2 plays important roles in diverse nuclear functions, including RNA metabolism and maintenance of nuclear architecture. PMID- 11927595 TI - Membrane association domains in Ca2+-dependent activator protein for secretion mediate plasma membrane and dense-core vesicle binding required for Ca2+ dependent exocytosis. AB - Ca2+-dependent activator protein for secretion (CAPS) is a cytosolic protein essential for the Ca2+-dependent fusion of dense-core vesicles (DCVs) with the plasma membrane and the regulated secretion of a subset of neurotransmitters. The mechanism by which CAPS functions in exocytosis and the means by which it associates with target membranes are unknown. We identified two domains in CAPS with distinct membrane-binding properties that were each essential for CAPS activity in regulated exocytosis. The first of these, a centrally located pleckstrin homology domain, exhibited three properties: charge-based binding to acidic phospholipids, binding to plasma membrane but not DCV membrane, and stereoselective binding to phosphatidylinositol 4,5-bisphosphate. Mutagenesis studies revealed that the former two properties but not the latter were essential for CAPS function. The central pleckstrin homology domain may mediate transient CAPS interactions with the plasma membrane during Ca2+-triggered exocytosis. The second membrane association domain comprising distal C-terminal sequences mediated CAPS targeting to and association with neuroendocrine DCVs. The CAPS C terminal domain was also essential for optimal activity in regulated exocytosis. The presence of two membrane association domains with distinct binding specificities may enable CAPS to bind both target membranes to facilitate DCV plasma membrane fusion. PMID- 11927594 TI - Identification of a novel non-structural maintenance of chromosomes (SMC) component of the SMC5-SMC6 complex involved in DNA repair. AB - Structural maintenance of chromosomes (SMC) proteins play central roles in chromosome organization and dynamics. They have been classified into six subtypes, termed SMC1 to SMC6, and function as heterodimer components of large protein complexes that also include several non-SMC proteins. The SMC2-SMC4 and SMC1-SMC3 complexes are also known as condensin and cohesin, respectively, but the recently identified SMC5 and SMC6 complex is less well characterized. Here, we report that NSE1 from Saccharomyces cerevisiae encodes a novel non-SMC component of the SMC5(Yol034wp)-SMC6(Rhc18p) complex corresponding to the 2-3-MDa molecular mass. Nse1p is essential for cell proliferation and localizes primarily in the nucleus. nse1 mutants are highly sensitive to DNA-damaging treatments and exhibit abnormal cellular morphologies, suggesting aberrant mitosis as a terminal morphological phenotype. These results are consistent with the reported features of the Schizosaccharomyces pombe SMC6 gene, rad18, which is thought to be involved in recombinational DNA repair. We conclude that Nse1p and the SMC5-SMC6 heterodimer together form a high molecular mass complex that is conserved in eukaryotes and required for both DNA repair and proliferation. PMID- 11927596 TI - Biochemical characterization of two analogues of the apoptosis-linked gene 2 protein in Dictyostelium discoideum and interaction with a physiological partner in mammals, murine Alix. AB - Two homologues, Dd-ALG-2a and Dd-ALG-2b, of the mammalian calcium-binding protein ALG-2 (apoptosis-linked gene 2) have been characterized in the cellular slime mold Dictyostelium discoideum. Fluorescence titrations showed that both proteins bind calcium ions with affinities (Ca2+)(0.5) of 30 and 450 microm, respectively, at sites specific to calcium. Calcium ion binding resulted in changes of conformation associated with the unmasking of hydrophobic regions of the proteins. Surface plasmon resonance analysis showed that Dd-ALG-2a homodimers formed (K(D) of 1 microm) at calcium ion concentrations similar to those necessary for Ca2+-induced conformational changes. Deletion of the hydrophobic N terminal sequence or EF-hand 5 of Dd-ALG-2a prevented dimerization. The Dd-ALG-2b homodimer was not detected, and the Dd-ALG-2a/2b heterodimer formed only when Dd ALG-2b was the immobilized partner. Murine Alix formed a heterodimer (K(D) = 0.6 microm) with Dd-ALG-2a but not with Dd-ALG-2b, and the interaction strictly depended upon calcium ions. The DeltaNter construct of Dd-ALG-2a lost its interaction capacity with mouse Alix. The genes encoding both proteins, Dd-alg-2a and -2b, were expressed in growing cells. The levels of mRNA were at a maximum during aggregation (4-8 h) and decreased rapidly thereafter. In contrast, the levels of proteins remained fairly stable. Dd-ALG-2a and Dd-ALG-2b were found to be dispensable for growth and development, based on the finding that single Dd alg2a- or Dd-alg-2b- and double Dd-alg2a-/Dd-alg-2b- mutant cell lines showed normal growth in axenic medium or on bacterial lawns and exhibited unaltered development. PMID- 11927597 TI - Direct interaction between uracil-DNA glycosylase and a proliferating cell nuclear antigen homolog in the crenarchaeon Pyrobaculum aerophilum. AB - Proliferating cell nuclear antigen (PCNA) acts as a sliding clamp on duplex DNA. Its homologs, present in Eukarya and Archaea, are part of protein complexes that are indispensable for DNA replication and DNA repair. In Eukarya, PCNA is known to interact with more than a dozen different proteins, including a human major nuclear uracil-DNA glycosylase (hUNG2) involved in immediate postreplicative repair. In Archaea, only three classes of PCNA-binding proteins have been reported previously: replication factor C (the PCNA clamp loader), family B DNA polymerase, and flap endonuclease. In this study, we report a direct interaction between a uracil-DNA glycosylase (Pa-UDGa) and a PCNA homolog (Pa-PCNA1), both from the hyperthermophilic crenarchaeon Pyrobaculum aerophilum (T(opt) = 100 degrees C). We demonstrate that the Pa-UDGa-Pa-PCNA1 complex is thermostable, and two hydrophobic amino acid residues on Pa-UDGa (Phe(191) and Leu(192)) are shown to be crucial for this interaction. It is interesting to note that although Pa UDGa has homologs throughout the Archaea and bacteria, it does not share significant sequence similarity with hUNG2. Nevertheless, our results raise the possibility that Pa-UDGa may be a functional analog of hUNG2 for PCNA-dependent postreplicative removal of misincorporated uracil. PMID- 11927598 TI - Species specificity of simian virus 40 DNA replication in vitro requires multiple functions of human DNA polymerase alpha. AB - Human cell extracts support the replication of SV40 DNA, whereas mouse cell extracts do not. Species specificity is determined at the level of initiation of DNA replication, and it was previously found that this requires the large subunit, p180, of DNA polymerase alpha-primase to be of human origin. Furthermore, a functional interaction between SV40 large T antigen (TAg) and p180 is essential for viral DNA replication. In this study we determined that the N terminal regions of human p180, which contain the TAg-binding sites, can be replaced with those of murine origin without losing the ability to support SV40 DNA replication in vitro. The same substitutions do not prevent SV40 TAg from stimulating the activity of DNA polymerase alpha-primase on single-stranded DNA in the presence of replication protein A. Furthermore, biophysical studies show that the interactions of human and murine DNA polymerase alpha-primase with SV40 TAg are of a similar magnitude. These studies strongly suggest that requirement of SV40 DNA replication for human DNA polymerase alpha depends neither on the TAg binding site being of human origin nor on the strength of the binary interaction between SV40 TAg and DNA polymerase alpha-primase but rather on sequences in the C-terminal region of human p180. PMID- 11927599 TI - Potassium functionally replaces the second lysine of the KMSKS signature sequence in human tyrosyl-tRNA synthetase. AB - Unlike their bacterial homologues, a number of eukaryotic tyrosyl-tRNA synthetases require potassium to catalyze the aminoacylation reaction. In addition, the second lysine in the class I-specific KMSKS signature motif is absent from all known eukaryotic tyrosyl-tRNA synthetase sequences, except those of higher plants. This lysine, which is the most highly conserved residue in the class I aminoacyl-tRNA synthetase family, has been shown to interact with the pyrophosphate moiety of the ATP substrate in the Bacillus stearothermophilus tyrosyl-tRNA synthetase. Equilibrium dialysis and pre-steady-state kinetic analyses were used to determine the role that potassium plays in the tyrosine activation reaction in the human tyrosyl-tRNA synthetase and whether it can be replaced by any of the other alkali metals. Kinetic analyses indicate that potassium interacts with the pyrophosphate moiety of ATP, stabilizing the E.Tyr.ATP and E.[Tyr-ATP] complexes by 2.3 and 4.3 kcal/mol, respectively. Potassium also appears to stabilize the asymmetric conformation of the human tyrosyl-tRNA synthetase dimer by 0.7 kcal/mol. Rubidium is the only other alkali metal that can replace potassium in catalyzing tyrosine activation, although the forward rate constant is half of that observed when potassium is present. The above results are consistent with the hypothesis that potassium functionally replaces the second lysine in the KMSKS signature sequence. Possible implications of these results with respect to the design of antibiotics that target bacterial aminoacyl-tRNA synthetases are discussed. PMID- 11927600 TI - Intrinsic sensitivity of Kir1.1 (ROMK) to glibenclamide in the absence of SUR2B. Implications for the identity of the renal ATP-regulated secretory K+ channel. AB - The precise molecular identity of the renal ATP-regulated secretory K+ channel is still a matter of some controversy. The inwardly rectifying K+ channel, Kir1.1 (ROMK) appears to form the pore of the channel, and mutations in Kir1.1 are responsible for Bartter syndrome. The native channel is sensitive to inhibition by the sulfonylurea glibenclamide, and it has been proposed that an accessory protein is required to confer glibenclamide sensitivity to Kir1.1. Several recent studies have suggested that the native channel is composed of the splice variant Kir1.1b (ROMK2) and the sulfonylurea receptor isoform SUR2B and that there is a direct physical interaction between these subunits. In this study, we have monitored the interaction between Kir1.1b and SUR2B. We find that SUR2B reaches the plasma membrane when coexpressed with Kir6.1 or Kir6.2 but not when coexpressed with Kir1.1b. Furthermore, we find that Kir1.1b exhibits an intrinsic sensitivity to inhibition by glibenclamide with an affinity similar to the native channel. These results demonstrate that SUR2B does not traffic to the membrane in the presence of Kir1.1b and is not required to confer glibenclamide sensitivity to Kir1.1b. This has important implications for the presumed structure of the renal ATP-regulated secretory K+ channel. PMID- 11927601 TI - Fibroblast growth factor 18 influences proximal programming during lung morphogenesis. AB - The structure and functions of the airways of the lung change dramatically along their lengths. Large-diameter conducting airways are supported by cartilaginous rings and smooth muscle tissue and are lined by ciliated and secretory epithelial cells that are involved in mucociliary clearance. Smaller peripheral airways formed during branching morphogenesis are lined by cuboidal and squamous cells that facilitate gas exchange to a network of fine capillaries. The factors that mediate formation of these changing cell types and structures along the length of the airways are unknown. We report here that conditional expression of fibroblast growth factor (FGF)-18 in epithelial cells of the developing lung caused the airway to adopt structural features of proximal airways. Peripheral lung tubules were markedly diminished in numbers, whereas the size and extent of conducting airways were increased. Abnormal smooth muscle and cartilage were found in the walls of expanded distal airways, which were accompanied by atypically large pulmonary blood vessels. Expression of proteins normally expressed in peripheral lung tubules, including SP-B and pro-SP-C, was inhibited. FGF-18 mRNA was detected in normal mouse lung in stromal cells surrounding proximal airway cartilage and in peripheral lung mesenchyme. Effects were unique to FGF-18 because expression of other members of the FGF family had different consequences. These data show that FGF-18 is capable of enhancing proximal and inhibiting peripheral programs during lung morphogenesis. PMID- 11927602 TI - Dynamics of human DNA topoisomerases IIalpha and IIbeta in living cells. AB - DNA topoisomerase (topo) II catalyses topological genomic changes essential for many DNA metabolic processes. It is also regarded as a structural component of the nuclear matrix in interphase and the mitotic chromosome scaffold. Mammals have two isoforms (alpha and beta) with similar properties in vitro. Here, we investigated their properties in living and proliferating cells, stably expressing biofluorescent chimera of the human isozymes. Topo IIalpha and IIbeta behaved similarly in interphase but differently in mitosis, where only topo IIalpha was chromosome associated to a major part. During interphase, both isozymes joined in nucleolar reassembly and accumulated in nucleoli, which seemed not to involve catalytic DNA turnover because treatment with teniposide (stabilizing covalent catalytic DNA intermediates of topo II) relocated the bulk of the enzymes from the nucleoli to nucleoplasmic granules. Photobleaching revealed that the entire complement of both isozymes was completely mobile and free to exchange between nuclear subcompartments in interphase. In chromosomes, topo IIalpha was also completely mobile and had a uniform distribution. However, hypotonic cell lysis triggered an axial pattern. These observations suggest that topo II is not an immobile, structural component of the chromosomal scaffold or the interphase karyoskeleton, but rather a dynamic interaction partner of such structures. PMID- 11927604 TI - The anti-apoptotic activity of XIAP is retained upon mutation of both the caspase 3- and caspase 9-interacting sites. AB - The X-linked mammalian inhibitor of apoptosis protein (XIAP) has been shown to bind several partners. These partners include caspase 3, caspase 9, DIABLO/Smac, HtrA2/Omi, TAB1, the bone morphogenetic protein receptor, and a presumptive E2 ubiquitin-conjugating enzyme. In addition, we show here that XIAP can bind to itself. To determine which of these interactions are required for it to inhibit apoptosis, we generated point mutant XIAP proteins and correlated their ability to bind other proteins with their ability to inhibit apoptosis. partial differential RING point mutants of XIAP were as competent as their full-length counterparts in inhibiting apoptosis, although impaired in their ability to oligomerize with full-length XIAP. Triple point mutants, unable to bind caspase 9, caspase 3, and DIABLO/HtrA2/Omi, were completely ineffectual in inhibiting apoptosis. However, point mutants that had lost the ability to inhibit caspase 9 and caspase 3 but retained the ability to inhibit DIABLO were still able to inhibit apoptosis, demonstrating that IAP antagonism is required for apoptosis to proceed following UV irradiation. PMID- 11927603 TI - Sequential tethering of Golgins and catalysis of SNAREpin assembly by the vesicle tethering protein p115. AB - p115 tethers coat protein (COP)I vesicles to Golgi membranes. The acidic COOH terminal domain of p115 links the Golgins, Giantin on COPI vesicles, to GM130 on Golgi membranes. We now show that a SNARE motif-related domain within p115 stimulates the specific assembly of endogenous Golgi SNAREpins containing the t SNARE, syntaxin 5. p115 catalyzes the construction of a cognate GOS-28-syntaxin-5 (v-/t-SNARE) complex by first linking the SNAREs to promote their direct interaction. These events are essential for NSF-catalyzed reassembly of postmitotic Golgi vesicles and tubules into mature cisternae. Staging experiments reveal that the linking of Golgins precedes SNAREpin assembly. Thus, p115 coordinates sequential tethering and docking of COPI vesicles by first using long tethers (Golgins) and then short tethers (SNAREs). PMID- 11927605 TI - Muscle-specific RING finger-1 interacts with titin to regulate sarcomeric M-line and thick filament structure and may have nuclear functions via its interaction with glucocorticoid modulatory element binding protein-1. AB - The COOH-terminal A168-170 region of the giant sarcomeric protein titin interacts with muscle-specific RING finger-1 (MURF-1). To investigate the functional significance of this interaction, we expressed green fluorescent protein fusion constructs encoding defined fragments of titin's M-line region and MURF-1 in cardiac myocytes. Upon expression of MURF-1 or its central region (containing its titin-binding site), the integrity of titin's M-line region was dramatically disrupted. Disruption of titin's M-line region also resulted in a perturbation of thick filament components, but, surprisingly, not of the NH2-terminal or I-band regions of titin, the Z-lines, or the thin filaments. This specific phenotype also was caused by the expression of titin A168-170. These data suggest that the interaction of titin with MURF-1 is important for the stability of the sarcomeric M-line region.MURF-1 also binds to ubiquitin-conjugating enzyme-9 and isopeptidase T-3, enzymes involved in small ubiquitin-related modifier-mediated nuclear import, and with glucocorticoid modulatory element binding protein-1 (GMEB-1), a transcriptional regulator. Consistent with our in vitro binding data implicating MURF-1 with nuclear functions, endogenous MURF-1 also was detected in the nuclei of some myocytes. The dual interactions of MURF-1 with titin and GMEB 1 may link myofibril signaling pathways (perhaps including titin's kinase domain) with muscle gene expression. PMID- 11927606 TI - Muscle-specific expression of insulin-like growth factor I counters muscle decline in mdx mice. AB - Duchenne muscular dystrophy is an X-linked degenerative disorder of muscle caused by the absence of the protein dystrophin. A major consequence of muscular dystrophy is that the normal regenerative capacity of skeletal muscle cannot compensate for increased susceptibility to damage, leading to repetitive cycles of degeneration-regeneration and ultimately resulting in the replacement of muscle fibers with fibrotic tissue. Because insulin-like growth factor I (IGF-I) has been shown to enhance muscle regeneration and protein synthetic pathways, we asked whether high levels of muscle-specific expression of IGF-I in mdx muscle could preserve muscle function in the diseased state. In transgenic mdx mice expressing mIgf-I (mdx:mIgf+/+), we showed that muscle mass increased by at least 40% leading to similar increases in force generation in extensor digitorum longus muscles compared with those from mdx mice. Diaphragms of transgenic mdx:mIgf+/+ exhibited significant hypertrophy and hyperplasia at all ages observed. Furthermore, the IGF-I expression significantly reduced the amount of fibrosis normally observed in diaphragms from aged mdx mice. Decreased myonecrosis was also observed in diaphragms and quadriceps from mdx:mIgf+/+ mice when compared with age-matched mdx animals. Finally, signaling pathways associated with muscle regeneration and protection against apoptosis were significantly elevated. These results suggest that a combination of promoting muscle regenerative capacity and preventing muscle necrosis could be an effective treatment for the secondary symptoms caused by the primary loss of dystrophin. PMID- 11927607 TI - Src-mediated coupling of focal adhesion kinase to integrin alpha(v)beta5 in vascular endothelial growth factor signaling. AB - Vascular endothelial growth factor (VEGF) promotes vascular permeability (VP) and neovascularization, and is required for development. We find that VEGF-stimulated Src activity in chick embryo blood vessels induces the coupling of focal adhesion kinase (FAK) to integrin alpha(v)beta5, a critical event in VEGF-mediated signaling and biological responsiveness. In contrast, FAK is constitutively associated with beta1 and beta3 integrins in the presence or absence of growth factors. In cultured endothelial cells, VEGF, but not basic fibroblast growth factor, promotes the Src-mediated phosphorylation of FAK on tyrosine 861, which contributes to the formation of a FAK/alpha(v)beta5 signaling complex. Moreover, formation of this FAK/alpha(v)beta5 complex is significantly reduced in pp60c-src deficient mice. Supporting these results, mice deficient in either pp60c-src or integrin beta5, but not integrin beta3, have a reduced VP response to VEGF. This FAK/alpha(v)beta5 complex was also detected in epidermal growth factor-stimulated epithelial cells, suggesting a function for this complex outside the endothelium. Our findings indicate that Src can coordinate specific growth factor and extracellular matrix inputs by recruiting integrin alpha(v)beta5 into a FAK containing signaling complex during growth factor-mediated biological responses. PMID- 11927608 TI - The Maguk protein, Pals1, functions as an adapter, linking mammalian homologues of Crumbs and Discs Lost. AB - Membrane-associated guanylate kinase (Maguk) proteins are scaffold proteins that contain PSD-95-Discs Large-zona occludens-1 (PDZ), Src homology 3, and guanylate kinase domains. A subset of Maguk proteins, such as mLin-2 and protein associated with Lin-7 (Pals)1, also contain two L27 domains: an L27C domain that binds mLin 7 and an L27N domain of unknown function. Here, we demonstrate that the L27N domain targets Pals1 to tight junctions by binding to a PDZ domain protein, Pals1 associated tight junction (PATJ) protein, via a unique Maguk recruitment domain. PATJ is a homologue of Drosophila Discs Lost, a protein that is crucial for epithelial polarity and that exists in a complex with the apical polarity determinant, Crumbs. PATJ and a human Crumbs homologue, CRB1, colocalize with Pals1 to tight junctions, and CRB1 interacts with PATJ albeit indirectly via binding the Pals1 PDZ domain. In agreement, we find that a Drosophila homologue of Pals1 participates in identical interactions with Drosophila Crumbs and Discs Lost. This Drosophila Pals1 homologue has been demonstrated recently to represent Stardust, a crucial polarity gene in Drosophila. Thus, our data identifies a new multiprotein complex that appears to be evolutionarily conserved and likely plays an important role in protein targeting and cell polarity. PMID- 11927609 TI - PECAM-1 (CD31) regulates a hydrogen peroxide-activated nonselective cation channel in endothelial cells. AB - Hydrogen peroxide (H2O2) released by neutrophils is an important mediator of endothelial cell (EC) injury and vascular inflammation via its effect on EC-free Ca2+, [Ca2+]i. Although the underlying mechanisms are not well understood, platelet endothelial cell adhesion molecule (PECAM)-1/CD-31 is a critical modulator of neutrophil-EC transmigration. PECAM-1 is also known to regulate EC calcium signals and to undergo selective tyrosine phosphorylation. Here, we report that PECAM-1 molecules transduce EC responses to hydrogen peroxide. In human umbilical vein EC and REN cells (a PECAM-1-negative EC-like cell line) stably transfected with PECAM-1 (RHP), noncytolytic H2O2 exposure (100-200 microM H2O2) activated a calcium-permeant, nonselective cation current, and a transient rise in [Ca2+]i of similar time course. Neither response was observed in untransfected REN cells, and H2O2-evoked cation current was ablated in REN cells transfected with PECAM-1 constructs mutated in the cytoplasmic tyrosine containing domain. The PECAM-dependent H2O2 current was inhibited by dialysis of anti-PECAM-1 cytoplasmic domain antibodies, required Src family tyrosine kinase activity, was independent of inositol trisphosphate receptor activation, and required only an intact PECAM-1 cytoplasmic domain. PECAM-1-dependent H2O2 currents and associated [Ca2+]i transients may play a significant role in regulating neutrophil-endothelial interaction, as well as in oxidant-mediated endothelial response and injury. PMID- 11927610 TI - Mixed signals in heart failure: cancer rules. PMID- 11927612 TI - Dysregulation of Met receptor tyrosine kinase activity in invasive tumors. PMID- 11927613 TI - Controlling the controls: GAD65 autoreactive T cells in type 1 diabetes. PMID- 11927611 TI - Invasive growth: from development to metastasis. PMID- 11927614 TI - Regulation of herpes simplex virus gamma(1)34.5 expression and oncolysis of diffuse liver metastases by Myb34.5. AB - Myb34.5 is a herpes simplex virus 1 (HSV-1) mutant deleted in the gene for ribonucleotide reductase (ICP6). It also carries a version of gamma(1)34.5 (a viral gene product that promotes the dephosphorylation of eIF-2alpha) that is under control of the E2F-responsive cellular B-myb promoter, rather than of its endogenous promoter. Myb34.5 replication in tumor cells results in their destruction (oncolysis). gamma(1)34.5 expression by HSV-1 subverts an important cell defense mechanism against viral replication by preventing shutoff of protein synthesis after viral infection. Infection of colon carcinoma cells with Myb34.5 results in greater eIF-2alpha dephosphorylation and viral replication compared with infection with HSV-1 mutants completely defective in gamma(1)34.5 expression. In contrast, infection of normal hepatocytes with Myb34.5 results in low levels of eIF-2alpha dephosphorylation and viral replication that are similar to those observed with HSV-1 mutants completely defective in gamma(1)34.5 and ICP6. When administered intravascularly into mice with diffuse liver metastases, Myb34.5 has greater antineoplastic activity than HSV-1 mutants with completely defective gamma(1)34.5 expression and more restricted biodistribution compared with HSV-1 mutants with wild-type gamma(1)34.5 expression. Myb34.5 displays reduced virulence and toxicity compared to HSV-1 mutants with wild-type gamma(1)34.5 expression. Portal venous administration of Myb34.5 significantly reduces liver tumor burden in and prolongs the life of mice with diffuse liver metastases. Preexisting Ab's to HSV-1 do not reduce the antitumor efficacy of Myb34.5 in vivo. PMID- 11927615 TI - The prostaglandin receptor EP4 suppresses colitis, mucosal damage and CD4 cell activation in the gut. AB - We used mice deficient in each of the eight types and subtypes of prostanoid receptors and examined the roles of prostanoids in dextran sodium sulfate-induced (DSS-induced) colitis. Among the prostanoid receptor-deficient mice, only EP4 deficient mice and not mice deficient in either DP, EP1, EP2, EP3, FP, IP, or TP developed severe colitis with 3% DSS treatment, which induced only marginal colitis in wild-type mice. This phenotype was mimicked in wild-type mice by administration of an EP4-selective antagonist (AE3-208). The EP4 deficiency impaired mucosal barrier function and induced epithelial loss, crypt damage, and aggregation of neutrophils and lymphocytes in the colon. Conversely, administration of an EP4-selective agonist (AE1-734) to wild-type mice ameliorated severe colitis normally induced with 7% DSS, while that of AE3-208 suppressed recovery from colitis and induced significant proliferation of CD4+ T cells. In vitro AE3-208 enhanced and AE1-734 suppressed the proliferation and Th1 cytokine production of lamina propria mononuclear cells from the colon. DNA microarray analysis revealed elevated expression of genes associated with immune response and reduced expression of genes with mucosal repair and remodeling in the colon of EP4-deficient mice. We conclude that EP4 maintains intestinal homeostasis by keeping mucosal integrity and downregulating immune response. PMID- 11927616 TI - GAD65-reactive T cells are activated in patients with autoimmune type 1a diabetes. AB - Insulin-dependent type 1 diabetes is an autoimmune disease mediated by T lymphocytes recognizing pancreatic islet cell antigens. Glutamic acid decarboxylase 65 (GAD65) appears to be an important autoantigen in the disease. However, T cells from both patients with type 1 diabetes and healthy subjects vigorously proliferate in response to GAD65 stimulation ex vivo, leading us to postulate that the critical event in the onset of human diabetes is the activation of autoreactive T cells. Thus, we investigated whether GAD65-reactive T cells in patients with diabetes functioned as previously activated memory T cells, no longer requiring a second, costimulatory signal for clonal expansion. We found that in patients with new-onset type 1 diabetes, GAD65-reactive T cells were strikingly less dependent on CD28 and B7-1 costimulation to enter into cell cycle and proliferate than were equivalent cells derived from healthy controls. We hypothesize that these autoreactive T cells have been activated in vivo and have differentiated into memory cells, suggesting a pathogenic role in type 1 diabetes. In addition, we observed different effects with selective blockade of either B7-1 or B7-2 molecules; B7-1 appears to deliver a negative signal by engaging CTLA-4, while B7-2 engagement of CD28 upregulates T cell proliferation and cytokine secretion. PMID- 11927617 TI - Critical role for a high-affinity chemokine-binding protein in gamma-herpesvirus induced lethal meningitis. AB - Chemokines are involved in recruitment and activation of hematopoietic cells in sites of infection and inflammation. The M3 gene of the gamma-herpesvirus gammaHV68 encodes an abundant secreted protein that binds CC chemokines with high affinity. We report here that this gene is essential for efficient induction of lethal meningitis by gammaHV68. An M3 mutant gammaHV68 (gammaHV68-M3.stop) was 100-fold less virulent than wild-type or marker rescue control (gammaHV68-M3.MR) viruses after intracerebral inoculation. After intracerebral inoculation, gammaHV68-M3.stop grew to lower titers than gammaHV68 or gammaHV68-M3.MR in the brain but spread to and grew normally in the spleen and lung. Expression of several CC chemokines was significantly induced in the CNS by gammaHV68 infection. Consistent with M3 acting by blockade of CC chemokine action, gammaHV68 induced a neutrophilic meningeal inflammatory infiltrate, while gammaHV68-M3.stop induced an infiltrate in which lymphocytes and macrophages predominated. In contrast to the important role of M3 in lethal meningitis, M3 was not required for establishment or reactivation from latent infection or induction of chronic arteritis. These data suggest a role for chemokines in the protection of the nervous system from viral infection and that the M3 protein acts in a tissue-specific fashion during acute but not chronic gammaHV68 infection to limit CC chemokine-induced inflammatory responses. PMID- 11927618 TI - Hypothalamic Y2 receptors regulate bone formation. AB - Neuropeptide Y (NPY) is a downstream modulator of leptin action, possibly at the level of the arcuate nucleus where NPY neurons are known to express both leptin receptors and Y2 receptors. In addition to the well-described role of NPY and leptin in energy balance and obesity, intracerebroventricular administration of NPY or leptin also causes bone loss. Here we show that Y2 receptor-deficient mice have a twofold increase in trabecular bone volume as well as greater trabecular number and thickness compared with control mice. We also demonstrate that central Y2 receptors are crucial for this process, since selective deletion of hypothalamic Y2 receptors in mature conditional Y2 knockout mice results in an identical increase in trabecular bone volume within 5 weeks. This hypothalamus specific Y2 receptor deletion stimulates osteoblast activity and increases the rate of bone mineralization and formation, with no effect on osteoblast or osteoclast surface measurements. The lack of any changes in plasma total calcium, leptinemia, or hypothalamo-pituitary-corticotropic, -thyrotropic, -somatotropic, or -gonadotropic output suggests that Y2 receptors do not modulate bone formation by humoral mechanisms, and that alteration of autonomic function through hypothalamic Y2 receptors may play a key role in a major central regulatory circuit of bone formation. PMID- 11927619 TI - The melanin-concentrating hormone receptor 1, a novel target of autoantibody responses in vitiligo. AB - Vitiligo is a common depigmenting disorder resulting from the loss of melanocytes in the skin. The pathogenesis of the disease remains obscure, although autoimmune mechanisms are thought to be involved. Indeed, autoantibodies and autoreactive T lymphocytes that target melanocytes have been reported in some vitiligo patients. The objective of this study was to identify pigment cell antigens that are recognized by autoantibodies in vitiligo. Using IgG from vitiligo patients to screen a melanocyte cDNA phage-display library, we identified the melanin concentrating hormone receptor 1 (MCHR1) as a novel autoantigen related to this disorder. Immunoreactivity against the receptor was demonstrated in vitiligo patient sera by using radiobinding assays. Among sera from healthy controls and from patients with autoimmune disease, none exhibited immunoreactivity to MCHR1, indicating a high disease specificity for Ab's against the receptor. Inhibition of MCH binding to its receptor by IgG from vitiligo patients was also shown. PMID- 11927620 TI - IL-7 inhibits fibroblast TGF-beta production and signaling in pulmonary fibrosis. AB - Based on studies by our group and others, we hypothesized that IL-7 may possess antifibrotic activities in an IFN-gamma-dependent and independent manner. Here, we have evaluated the antifibrotic therapeutic potential of IL-7 in both in vitro and in vivo pulmonary fibrosis models. IL-7 inhibited both TGF-beta production and signaling in fibroblasts and required an intact JAK1/STAT1 signal transduction pathway. IL-7-mediated inhibition of TGF-beta signaling was found to be associated with an increase in Smad7, a major inhibitory regulator in the SMAD family. In the presence of IL-7, Smad7 dominant negative fibroblasts restored TGF beta-induced collagen synthesis, indicating that an IL-7-mediated increase in Smad7 suppressed TGF-beta signaling. Consistent with these in vitro findings, recombinant IL-7 decreased bleomycin-induced pulmonary fibrosis in vivo, independent of IFN-gamma. The antifibrotic activities of IL-7 merit further basic and clinical investigation for the treatment of pulmonary fibrosis. PMID- 11927621 TI - P-selectin glycoprotein ligand-1-deficient mice have impaired leukocyte tethering to E-selectin under flow. AB - P-selectin glycoprotein ligand-1 (PSGL-1) mediates rolling of leukocytes on P selectin under flow. The glycoproteins that enable leukocyte tethering to or rolling on E-selectin are not known. We used gene targeting to prepare PSGL-1 deficient (PSGL-1-/-) mice, which were healthy but had moderately elevated total blood leukocytes. Fluid-phase E-selectin bound to approximately 70% fewer sites on PSGL-1-/- than PSGL-1+/+ neutrophils. Compared with PSGL-1+/+ leukocytes, significantly fewer PSGL-1-/- leukocytes rolled on E-selectin in vitro, because their initial tethering to E-selectin was impaired. The residual cells that tethered rolled with the same shear resistance and velocities as PSGL-1+/+ leukocytes. Compared with PSGL-1+/+ mice, significantly fewer PSGL-1-/- leukocytes rolled on E-selectin in TNF-alpha-treated venules of cremaster muscle in which P-selectin function was blocked by an mAb. The residual PSGL-1-/- leukocytes that tethered rolled with slow velocities equivalent to those of PSGL 1+/+ leukocytes. These results reveal a novel function for PSGL-1 in tethering leukocytes to E-selectin under flow. PMID- 11927622 TI - IL-15, a survival factor for kidney epithelial cells, counteracts apoptosis and inflammation during nephritis. AB - IL-15, a T cell growth factor, has been linked to exacerbating autoimmune diseases and allograft rejection. To test the hypothesis that IL-15-deficient (IL 15-/-) mice would be protected from T cell-dependent nephritis, we induced nephrotoxic serum nephritis (NSN) in IL-15-/- and wild-type (IL-15+/+) C57BL/6 mice. Contrary to our expectations, IL-15 protects the kidney during this T cell dependent immunologic insult. Tubular, interstitial, and glomerular pathology and renal function are worse in IL-15-/- mice during NSN. We detected a substantial increase in tubular apoptosis in IL-15-/- kidneys. Moreover, macrophages and CD4 T cells are more abundant in the interstitia and glomeruli in IL-15-/- mice. This led us to identify several mechanisms responsible for heightened renal injury in the absence of IL-15. We now report that IL-15 and the IL-15 receptor (alpha, beta, gamma chains) are constitutively expressed in normal tubular epithelial cells (TECs). IL-15 is an autocrine survival factor for TECs. TEC apoptosis induced with anti-Fas or actinomycin D is substantially greater in IL-15-/- than in wild-type TECs. Moreover, IL-15 decreases the induction of a nephritogenic chemokine, MCP-1, that attracts leukocytes into the kidney during NSN. Taken together, we suggest that IL-15 is a therapeutic for tubulointerstitial and glomerular kidney diseases. PMID- 11927623 TI - Bile acid-activated nuclear receptor FXR suppresses apolipoprotein A-I transcription via a negative FXR response element. AB - Serum levels of HDL are inversely correlated with the risk of coronary heart disease. The anti-atherogenic effect of HDL is partially mediated by its major protein constituent apoA-I. In this study, we identify bile acids that are activators of the nuclear receptor farnesoid X receptor (FXR) as negative regulators of human apoA-I expression. Intrahepatocellular accumulation of bile acids, as seen in patients with progressive familial intrahepatic cholestasis and biliary atresia, was associated with diminished apoA-I serum levels. In human apoA-I transgenic mice, treatment with the FXR agonist taurocholic acid strongly decreased serum concentrations and liver mRNA levels of human apoA-I, which was associated with reduced serum HDL levels. Incubation of human primary hepatocytes and hepatoblastoma HepG2 cells with bile acids resulted in a dose-dependent downregulation of apoA-I expression. Promoter mutation analysis and gel-shift experiments in HepG2 cells demonstrated that bile acid-activated FXR decreases human apoA-I promoter activity by a negative FXR response element mapped to the C site. FXR bound this site and repressed transcription in a manner independent of retinoid X receptor. The nonsteroidal synthetic FXR agonist GW4064 likewise decreased apoA-I mRNA levels and promoter activity in HepG2 cells. PMID- 11927624 TI - Human sex hormone-binding globulin variants associated with hyperandrogenism and ovarian dysfunction. AB - The access of testosterone and estradiol to target tissues is regulated by sex hormone-binding globulin (SHBG) in human blood. Serum SHBG levels are low in patients with hyperandrogenism, especially in association with polycystic ovarian syndrome (PCOS) and in individuals at risk for diabetes and heart disease. Here, we identify SHBG coding region variations from a compound heterozygous patient who presented with severe hyperandrogenism during pregnancy. Serum SHBG levels in this patient measured 2 years after her pregnancy were exceptionally low, and her non-protein-bound testosterone concentrations greatly exceeded the normal reference range. A single-nucleotide polymorphism within the proband's maternally derived SHBG allele encodes a missense mutation, P156L, which allows for normal steroid ligand binding but causes abnormal glycosylation and inefficient secretion of SHBG. This polymorphism was identified in four other patients with either PCOS, ioiopathic hirsutism, or ovarian failure. The proband's paternal SHBG allele carries a single-nucleotide deletion within exon 8, producing a reading-frame shift within the codon for E326 and a premature termination codon. CHO cells transfected with a SHBG cDNA carrying this mutation fail to secrete the predicted truncated form of SHBG. To our knowledge, these are the first examples of human SHBG variants linked to hyperandrogenism and ovarian dysfunction. PMID- 11927626 TI - c-Jun NH(2)-terminal kinase (JNK)1 and JNK2 have distinct roles in CD8(+) T cell activation. AB - The c-Jun NH(2)-terminal kinase (JNK) signaling pathway is induced by cytokines and stress stimuli and is implicated in cell death and differentiation, but the specific function of this pathway depends on the cell type. Here we examined the role of JNK1 and JNK2 in CD8(+) T cells. Unlike CD4(+) T cells, the absence of JNK2 causes increased interleukin (IL)-2 production and proliferation of CD8(+) T cells. In contrast, JNK1-deficient CD8(+) T cells are unable to undergo antigen stimulated expansion in vitro, even in the presence of exogenous IL-2. The hypoproliferation of these cells is associated with impaired IL-2 receptor alpha chain (CD25) gene and cell surface expression. The reduced level of nuclear activating protein 1 (AP-1) complexes in activated JNK1-deficient CD8(+) T cells can account for the impaired IL-2 receptor alpha chain gene expression. Thus, JNK1 and JNK2 play different roles during CD8(+) T cell activation and these roles differ from those in CD4(+) T cells. PMID- 11927625 TI - c-Jun NH(2)-terminal kinase (JNK)1 and JNK2 signaling pathways have divergent roles in CD8(+) T cell-mediated antiviral immunity. AB - c-Jun NH(2)-terminal kinases (JNK) play important roles in T helper cell (Th) proliferation, differentiation, and maintenance of Th1/Th2 polarization. To determine whether JNKs are involved in antiviral T cell immunity, and whether JNK1 and JNK2 bear biological differences, we investigated the immune responses of JNK1-deficient and JNK2-deficient mice to lymphocytic choriomeningitis virus (LCMV). After LCMV infection, wild-type (JNK(+/+)) mice had a 5- to 10-fold increase in splenic CD8(+) T cells. In contrast, infected JNK1(-/-) mice showed a significantly lower virus-specific CD8(+) T cell expansion. However, JNK1(-/-) mice cleared LCMV infection with similar kinetics as JNK(+/+) mice. Splenic T cells from LCMV-infected JNK1(-/-) animals produced interferon gamma after stimulation with viral peptides. However, fewer JNK1(-/-) T cells acquired an activated phenotype (CD44(hi)) and more JNK1(-/-)CD8(+)CD44(hi) cells underwent apoptosis than JNK(+/+) cells at the peak of the primary response. In contrast, LCMV-infected JNK2(-/-) mice generated more virus-specific CD8(+) T cells than JNK(+/+) mice. These results indicate that JNK1 and JNK2 signal pathways have distinct roles in T cell responses during a viral infection. JNK1 is involved in survival of activated T cells during immune responses, and JNK2 plays a role in control of CD8(+) T cell expansion in vivo. PMID- 11927627 TI - Pivotal role of KARAP/DAP12 adaptor molecule in the natural killer cell-mediated resistance to murine cytomegalovirus infection. AB - Natural killer (NK) cells are major contributors to early defense against infections. Their effector functions are controlled by a balance between activating and inhibiting signals. To date, however, the involvement of NK cell activating receptors and signaling pathways in the defense against pathogens has not been extensively investigated. In mice, several NK cell activating receptors are coexpressed with and function through the immunoreceptor tyrosine-based activation motif (ITAM)-bearing molecule KARAP/DAP12. Here, we have analyzed the role of KARAP/DAP12 in the early antiviral response to murine cytomegalovirus (MCMV). In KARAP/DAP12 mutant mice bearing a nonfunctional ITAM, we found a considerable increase in viral titers in the spleen (30-40-fold) and in the liver (2-5-fold). These effects were attributed to NK cells. The formation of hepatic inflammatory foci appeared similar in wild-type and mutant mice, but the latter more frequently developed severe hepatitis with large areas of focal necrosis. Moreover, the percentage of hepatic NK cells producing interferon gamma was reduced by 56 +/- 22% in the absence of a functional KARAP/DAP12. This is the first study that shows a crucial role for a particular activating signaling pathway, in this case the one induced through KARAP/DAP12, in the NK cell mediated resistance to an infection. Our results are discussed in relation to recent reports demonstrating that innate resistance to MCMV requires the presence of NK cells expressing the KARAP/DAP12-associated receptor Ly49H. PMID- 11927628 TI - A natural killer T (NKT) cell developmental pathway iInvolving a thymus-dependent NK1.1(-)CD4(+) CD1d-dependent precursor stage. AB - The development of CD1d-dependent natural killer T (NKT) cells is poorly understood. We have used both CD1d/alpha-galactosylceramide (CD1d/alphaGC) tetramers and anti-NK1.1 to investigate NKT cell development in vitro and in vivo. Confirming the thymus-dependence of these cells, we show that CD1d/alphaGC tetramer-binding NKT cells, including NK1.1(+) and NK1.1(-) subsets, develop in fetal thymus organ culture (FTOC) and are completely absent in nude mice. Ontogenically, CD1d/alphaGC tetramer-binding NKT cells first appear in the thymus, at day 5 after birth, as CD4(+)CD8(-)NK1.1(-)cells. NK1.1(+) NKT cells, including CD4(+) and CD4(-)CD8(-) subsets, appeared at days 7-8 but remained a minor subset until at least 3 wk of age. Using intrathymic transfer experiments, CD4(+)NK1.1(-) NKT cells gave rise to NK1.1(+) NKT cells (including CD4(+) and CD4(-) subsets), but not vice-versa. This maturation step was not required for NKT cells to migrate to other tissues, as NK1.1(-) NKT cells were detected in liver and spleen as early as day 8 after birth, and the majority of NKT cells among recent thymic emigrants (RTE) were NK1.1(-). Further elucidation of this NKT cell developmental pathway should prove to be invaluable for studying the mechanisms that regulate the development of these cells. PMID- 11927629 TI - Clonality analysis of synchronous lesions of cervical carcinoma based on X chromosome inactivation polymorphism, human papillomavirus type 16 genome mutations, and loss of heterozygosity. AB - One of the most common forms of carcinoma in women, cervical invasive squamous cell carcinoma (CIC), often coexists with multiple lesions of cervical intraepithelial neoplasia (CIN). CIC and CIN show heterogeneity with respect to both histopathology and biology. To understand the causes, origin, and model of progression of cervical carcinoma, we assessed the clonality of a case with multiple synchronous lesions by analyzing X chromosome inactivation polymorphism, human papillomavirus type 16 (HPV16) sequence variation/mutations, and loss of heterozygosity (LOH). Microdissection was performed on 24 samples from this case, representing the entire lesional situation. The combination of different X chromosome inactivation patterns, two HPV16 point mutations, and LOH at three genomic microsatellite loci, led to the identification of five different "monoclonal" lesions (CIN II, CIN III, and invasive carcinoma nests) and five different "polyclonal" areas (CIN II and normal squamous epithelium). This finding indicated that CIC can originate from multiple precursor cells, from which some clones might progress via multiple steps, namely via CIN II and CIN III, whereas others might develop independently and possibly directly from the carcinoma precursor cells. Our results also supported the view that HPV16 as a "field factor" causes cervical carcinoma, which is probably promoted by the loss of chromosomal material as indicated by the LOH. PMID- 11927630 TI - Resident skin-specific gammadelta T cells provide local, nonredundant regulation of cutaneous inflammation. AB - The function of the intraepithelial lymphocyte (IEL) network of T cell receptor (TCR) gammadelta(+) (Vgamma5(+)) dendritic epidermal T cells (DETC) was evaluated by examining several mouse strains genetically deficient in gammadelta T cells (delta(-/-) mice), and in delta(-/-) mice reconstituted with DETC or with different gammadelta cell subpopulations. NOD.delta(-/-) and FVB.delta(-/-) mice spontaneously developed localized, chronic dermatitis, whereas interestingly, the commonly used C57BL/6.delta(-/-) strain did not. Genetic analyses indicated a single autosomal recessive gene controlled the dermatitis susceptibility of NOD.delta(-/-) mice. Furthermore, allergic and irritant contact dermatitis reactions were exaggerated in FVB.delta(-/-), but not in C57BL/6.delta(-/-) mice. Neither spontaneous nor augmented irritant dermatitis was observed in FVB.beta(-/ ) delta(-/-) mice lacking all T cells, indicating that alphabeta T cell-mediated inflammation is the target for gammadelta-mediated down-regulation. Reconstitution studies demonstrated that both spontaneous and augmented irritant dermatitis in FVB.delta(-/-) mice were down-regulated by Vgamma5(+) DETC, but not by epidermal T cells expressing other gammadelta TCRs. This study demonstrates that functional impairment at an epithelial interface can be specifically attributed to absence of the local TCR-gammadelta(+) IEL subset and suggests that systemic inflammatory reactions may more generally be subject to substantial regulation by local IELs. PMID- 11927631 TI - CD1d-restricted human natural killer T cells are highly susceptible to human immunodeficiency virus 1 infection. AB - Human natural killer (NK) T cells are unique T lymphocytes that express an invariant T cell receptor (TCR) Valpha24-Vbeta11 and have been implicated to play a role in various diseases. A subset of NKT cells express CD4 and hence are potential targets for human immunodeficiency virus (HIV)-1 infection. We demonstrate that both resting and activated human Valpha24(+) T cells express high levels of the HIV-1 coreceptors CCR5 and Bonzo (CXCR6), but low levels of CCR7, as compared with conventional T cells. Remarkably NKT cells activated with alpha-galactosylceramide (alpha-GalCer)-pulsed dendritic cells were profoundly more susceptible to infection with R5-tropic, but not X4-tropic, strains of HIV 1, compared with conventional CD4(+) T cells. Furthermore, resting CD4(+) NKT cells were also more susceptible to infection. After initial infection, HIV-1 rapidly replicated and depleted the CD4(+) subset of NKT cells. In addition, peripheral blood NKT cells were markedly and selectively depleted in HIV-1 infected individuals. Although the mechanisms of this decline are not clear, low numbers or absence of NKT cells may affect the course of HIV-1 infection. Taken together, our findings indicate that CD4(+) NKT cells are directly targeted by HIV-1 and may have a potential role during viral transmission and spread in vivo. PMID- 11927632 TI - The mechanism and significance of deletion of parasite-specific CD4(+) T cells in malaria infection. AB - It is thought that both helper and effector functions of CD4(+) T cells contribute to protective immunity to blood stage malaria infection. However, malaria infection does not induce long-term immunity and its mechanisms are not defined. In this study, we show that protective parasite-specific CD4(+) T cells were depleted after infection with both lethal and nonlethal species of rodent PLASMODIUM: It is further shown that the depletion is confined to parasite specific T cells because (a) ovalbumin (OVA)-specific CD4(+) T cells are not depleted after either malaria infection or direct OVA antigen challenge, and (b) the depletion of parasite-specific T cells during infection does not kill bystander OVA-specific T cells. A significant consequence of the depletion of malaria parasite-specific CD4(+) T cells is impaired immunity, demonstrated in mice that were less able to control parasitemia after depletion of transferred parasite-specific T cells. Using tumor necrosis factor (TNF)-RI knockout- and Fas deficient mice, we demonstrate that the depletion of parasite-specific CD4(+) T cells is not via TNF or Fas pathways. However, in vivo administration of anti interferon (IFN)-gamma antibody blocks depletion, suggesting that IFN-gamma is involved in the process. Taken together, these data suggest that long-term immunity to malaria infection may be affected by an IFN-gamma-mediated depletion of parasite-specific CD4(+) T cells during infection. This study provides further insight into the nature of immunity to malaria and may have a significant impact on approaches taken to develop a malaria vaccine. PMID- 11927633 TI - Epitope-specific evolution of human CD8(+) T cell responses from primary to persistent phases of Epstein-Barr virus infection. AB - Primary virus infection often elicits a large CD8(+) T cell response which subsequently contracts to a smaller memory T cell pool; the relationship between these two virus-specific populations is not well understood. Here we follow the human CD8(+) T cell response to Epstein-Barr virus (EBV) from its primary phase in infectious mononucleosis (IM) through to the persistent carrier state. Using HLA-A2.1 or B8 tetramers specific for four lytic cycle and three latent cycle epitopes, we find marked differences in the epitope-specific composition of the T cell populations between the two phases of infection. The primary response is dominated by lytic epitope specificities which are severely culled (and in one case extinguished) with resolution of the acute infection; in contrast latent epitope specificities are less abundant, if present at all, in acute IM but often then increase their percentage representation in the CD8 pool. Even comparing epitopes of the same type, the relative size of responses seen in primary infection does not necessarily correlate with that seen in the longer term. We also follow the evolution of phenotypic change in these populations and show that, from a uniform CD45RA(-)RO(+)CCR7(-) phenotype in IM, lytic epitope responses show greater reversion to a CD45RA(+)RO(-) phenotype whereas latent epitope responses remain CD45RA(-)RO(+) with a greater tendency to acquire CCR7. Interestingly these phenotypic distinctions reflect the source of the epitope as lytic or latent, and not the extent to which the response has been amplified in vivo. PMID- 11927634 TI - Role of p75 neurotrophin receptor in the neurotoxicity by beta-amyloid peptides and synergistic effect of inflammatory cytokines. AB - The neurodegenerative changes in Alzheimer's disease (AD) are elicited by the accumulation of beta-amyloid peptides (Abeta), which damage neurons either directly by interacting with components of the cell surface to trigger cell death signaling or indirectly by activating astrocytes and microglia to produce inflammatory mediators. It has been recently proposed that the p75 neurotrophin receptor (p75(NTR)) is responsible for neuronal damage by interacting with Abeta. By using neuroblastoma cell clones lacking the expression of all neurotrophin receptors or engineered to express full-length or various truncated forms of p75(NTR), we could show that p75(NTR) is involved in the direct signaling of cell death by Abeta via the function of its death domain. This signaling leads to the activation of caspases-8 and -3, the production of reactive oxygen intermediates and the induction of an oxidative stress. We also found that the direct and indirect (inflammatory) mechanisms of neuronal damage by Abeta could act synergistically. In fact, TNF-alpha and IL-1beta, cytokines produced by Abeta activated microglia, could potentiate the neurotoxic action of Abeta mediated by p75(NTR) signaling. Together, our results indicate that neurons expressing p75(NTR), mostly if expressing also proinflammatory cytokine receptors, might be preferential targets of the cytotoxic action of Abeta in AD. PMID- 11927635 TI - Major T cell progenitor activity in bone marrow-derived spleen colonies. AB - Common lymphoid progenitors (CLP) are generated in adult bone marrow (BM), but the intermediate steps leading to T cell commitment are unknown, and so is the site at which this commitment occurs. Here, we show that colonies arising in the spleen 12 days after BM injection harbor T cell precursors that are undetectable in BM. These precursors did not generate myeloid cells in vivo but repopulated the thymus and the peripheral T cell compartment much faster than did CLP. Two lineage negative (Lin(-)) subpopulations were distinguished, namely CD44(+) Thy1( ) cells still capable of natural killer generation and transient low-level B cell generation, and T cell-restricted CD44(-) Thy1(+) cells. At a molecular level, frequency of CD3epsilon and preTalpha mRNA was very different in each subset. Furthermore, only the CD44(-) Thy1(+) subset have initiated rearrangements in the T cell receptor beta locus. Thus, this study identifies extramedullary T cell progenitors and will allow easy approach to T cell commitment studies. PMID- 11927636 TI - Identification of the in vivo role of a viral bcl-2. AB - Many gamma-herpesviruses encode candidate oncogenes including homologues of host bcl-2 and cyclin proteins (v-bcl-2, v-cyclin), but the physiologic roles of these genes during infection are not known. We show for the first time in any virus system the physiologic role of v-bcl-2. A gamma-herpesvirus v-bcl-2 was essential for efficient ex vivo reactivation from latent infection, and for both persistent replication and virulence during chronic infection of immunocompromised (interferon [IFN]-gamma(-/-)) mice. The v-cyclin was also critical for the same stages in pathogenesis. Strikingly, while the v-bcl-2 and v-cyclin were important for chronic infection, these genes were not essential for viral replication in cell culture, viral replication during acute infection in vivo, establishment of latent infection, or virulence during acute infection. We conclude that v-bcl-2 and v-cyclin have important roles during latent and persistent gamma-herpesvirus infection and that herpesviruses encode genes with specific roles during chronic infection and disease, but not acute infection and disease. As gamma herpesviruses primarily cause human disease during chronic infection, these chronic disease genes may be important targets for therapeutic intervention. PMID- 11927637 TI - Zinc finger protein, Hzf, is required for megakaryocyte development and hemostasis. AB - Using an expression gene trapping strategy, we recently identified a novel gene, hematopoietic zinc finger (Hzf), which encodes a protein containing three C(2)H(2)-type zinc fingers that is predominantly expressed in megakaryocytes. Here, we have examined the in vivo function of Hzf by gene targeting and demonstrated that Hzf is essential for megakaryopoiesis and hemostasis in vivo. Hzf-deficient mice exhibited a pronounced tendency to rebleed and had reduced alpha-granule substances in both megakaryocytes and platelets. These mice also had large, faintly stained platelets, whereas the numbers of both megakaryocytes and platelets were normal. These results indicate that Hzf plays important roles in regulating the synthesis of alpha-granule substances and/or their packing into alpha-granules during the process of megakaryopoiesis. PMID- 11927638 TI - The development of murine plasmacytoid dendritic cell precursors is differentially regulated by FLT3-ligand and granulocyte/macrophage colony stimulating factor. AB - Plasmacytoid predendritic cells or type 1 interferon (IFN)-producing cells (IPCs) have recently been identified in mice. Although culture systems giving rise to different murine dendritic cell subsets have been established, the developmental regulation of murine plasmacytoid IPCs and the culture conditions leading to their generation remain unknown. Here we show that large numbers of over 40% pure CD11c(+)CD11b(-)B220(+)Gr-1(+) IPCs can be generated from mouse bone marrow cultures with FLT3-ligand. By contrast GM-CSF or TNF-alpha, which promote the generation of CD11c(+)CD11b(+)B220(-) myeloid DCs, block completely the development of IPCs. IPCs generated display similar features to human IPCs, such as the plasmacytoid morphology, the ability to produce large amounts of IFN-alpha in responses to herpes simplex virus, and the capacity to respond to ligands for Toll-like receptor 9 (TLR-9; CpG ODN 1668), but not to ligands for TLR-4 (lipopolysaccharide [LPS]). Unlike human IPCs which produce little IL-12p70, mouse IPCs produce IL-12p70 in response to CpG ODN 1668 and herpes simplex virus. This study demonstrates that the development of murine CD11c(+)CD11b(-)B220(+)Gr 1(+) IPCs and CD11c(+)CD11b(+)B220(-) myeloid DCs is differentially regulated by FLT3-ligand and granulocyte/macrophage colony-stimulating factor. Human IPCs and mouse IPCs display different ability to produce IL-12p70. Large numbers of mouse IPCs can now be obtained from total bone marrow culture. PMID- 11927639 TI - Persistent gamma-herpesvirus infections: what can we learn from an experimental mouse model? PMID- 11927640 TI - CBP and p300: versatile coregulators with important roles in hematopoietic gene expression. PMID- 11927641 TI - Differentiation of human basophils: an overview of recent advances and pending questions. AB - Basophils are rare, circulating leukocytes derived from hematopoietic CD34+ progenitors. The identification of cytokines promoting their development in vitro has led to substantial advances in understanding their differentiation process. An important role could be assigned to interleukin-3 (IL-3), which supports the maturation of hematopoietic progenitors into basophils in vitro and in vivo. In contrast to other myeloid lineages, a specific basophil growth factor has not yet been discovered. Furthermore, it is still unclear whether basophils possess a lineage-restricted progenitor or whether they share a common ancestor with mast cells (MC), eosinophils, or even megakaryocytes. Partial answers to these questions could be provided using in vitro culture systems or taking advantage of hematological disorders, such as chronic and acute myeloid leukemia (CML and AML), some myelodysplastic syndromes, and the very rare acute basophilic leukemia in which basophilic differentiation occurs. PMID- 11927642 TI - Interferon-alpha as an immunotherapeutic protein. AB - Interferon-alpha (IFN-alpha) has proven to be a clinically effective antiviral and antineoplastic therapeutic drug for more than 16 years. During this time, evidence from in vitro laboratory studies and the clinical arena has supported the concept that IFN-alpha is an immunotherapeutic drug. By regulating a diverse set of cytokines and their receptors, IFN-alpha is uniquely positioned to prime the host immune response and provide an effective antineoplastic- and antiviral immune response. IFN-alpha stimulates the innate cell-mediated response and then participates in the transition of the initial host innate response into an effective adaptive-immune response. IFN-alpha also drives the adaptive cell mediated CD8+ T-cell response and helps to maintain a CD4+ Th1-cell population balance for an effective antineoplastic and antiviral host defense. This review will describe the current state of knowledge of IFN-alpha as an immunoregulatory protein and address specific issues of IFN-alpha as an immunotherapeutic for antineoplastic and antiviral diseases. PMID- 11927643 TI - Rat monocyte-derived dendritic cells function and migrate in the same way as isolated tissue dendritic cells. AB - Dendritic cells (DC) are the most potent antigen-presenting cells and are therefore useful to induce immune responses against tumor cells in patients. DC can be generated in vitro from monocytes using GM-CSF and IL-4, the so-called monocyte-derived DC (MoDC). To achieve antitumor responses, MoDC must be able to migrate to the draining lymph nodes after injection to induce cytotoxic T cells. Therefore, we studied migration of MoDC in a rat model. Functional rat MoDC were generated from PVG-RT7B rats and injected subcutaneously into PVG rats. These rat strains differ only at one epitope of the leukocyte-common antigen, which can be recognized by the antibody His 41. The advantage is that migrated cells can be detected in the draining lymph nodes by staining sections with His 41+; thus, migration is not influenced by labeling procedures. Rat MoDC migrated to the T cell areas of the draining lymph nodes, just as isolated Langerhans cells or spleen DC do. In contrast, monocytes also migrated to the B-cell areas and the medulla. PMID- 11927644 TI - Effect of pentoxifylline on polarization and migration of human leukocytes. AB - Leukocyte polarization has a key role in the induction and effector phases of immune response. We assessed the effect of pentoxifylline (PTX) on the polarization and migration of human lymphocytes and neutrophils. A dose dependent, inhibitory effect on the polarization of lymphoid cells induced by chemokines or IL-15 was found. In addition, PTX interfered with the chemotaxis of peripheral blood T cells and T lymphoblasts. A similar effect was observed on the transendothelial migration of these cells. In addition, the polarization of neutrophils, its adherence to endothelium, and their transendothelial migration, induced by different stimuli, were inhibited by PTX. By contrast, this drug had only a mild effect on endothelial cells and a partial inhibition on the induction of ICAM-1 expression by TNF-alpha. The inhibitory effect of PTX on leukocyte polarization and extravasation may contribute significantly to the anti inflammatory and immunoregulatory activity of this drug. PMID- 11927645 TI - Differential expression of FIZZ1 and Ym1 in alternatively versus classically activated macrophages. AB - Alternatively activated macrophages (aaMphi) display molecular and biological characteristics that differ from those of classically activated macrophages (caMphi). Recently, we described an experimental model of murine trypanosomosis in which the early stage of infection of mice with a Trypanosoma brucei brucei variant is characterized by the development of caMphi, whereas in the late and chronic stages of infection, aaMphi develop. In the present study, we used suppression subtractive hybridization (SSH) to identify genes that are expressed differentially in aaMphi versus caMphi elicited during infection with this T. b. brucei variant. We show that FIZZ1 and Ym1 are induced strongly in in vivo- and in vitro-elicited aaMphi as compared with caMphi. Furthermore, we demonstrate that the in vivo induction of FIZZ1 and Ym1 in macrophages depends on IL-4 and that in vitro, IFN-gamma antagonizes the effect of IL-4 on the expression of FIZZ1 and Ym1. Collectively, these results open perspectives for new insights into the functional properties of aaMphi and establish FIZZ1 and Ym1 as markers for aaMphi. PMID- 11927646 TI - Ambient pCO2 modulates intracellular pH, intracellular oxidant generation, and interleukin-8 secretion in human neutrophils. AB - Although neutrophils are a critical component of the inflammatory process, their functional regulation is incompletely understood. Of note, although pCO2 varies physiologically and pathologically in the neutrophilic milieu, its affect on neutrophil biological processes is unresolved. We demonstrate here that neutrophils respond to hypo- and hypercarbia, (0.04% and 10%) by increasing and decreasing, respectively, intracellular oxidant production (basally and in response to opsonized Escherichia coli and phorbol esters). Further, hypo- and hypercarbia increase and decrease, respectively, the release of IL-8 from LPS stimulated cells; both effects are attenuated by the carbonic anhydrase inhibitor, acetazolamide. Anion exchange did not restore pH(i) under hypocarbic conditions, however partial restoration of pH(i) under hypercarbic conditions was achieved by Na+/H+ exchange and vacuolar ATPases. Abrogation of pCO2-induced changes in pH(i) prevented hypocarbia-induced generation of reactive oxidant species. These observations suggest that CO2 modifies neutrophil activity significantly by altering pH(i). PMID- 11927647 TI - Capacitative Ca2+ influx and activation of the neutrophil respiratory burst. Different regulation of plasma membrane- and granule-localized NADPH-oxidase. AB - The neutrophil NADPH-oxidase may be activated in the plasma membrane, resulting in release of oxygen metabolites extracellularly, or in the granule or phagosomal membranes, giving intracellular production of oxidants. An increase in [Ca2+]i mediated through binding of fMLF to its receptor is part of a signaling cascade that activates the plasma membrane-localized oxidase. In contrast, a rise in [Ca2+]i induced by a Ca2+ ionophore results in activation of the intracellular pool of oxidase. We mimicked fMLF-induced emptying of intracellular Ca2+ stores with thapsigargin. This induced a pronounced intracellular oxidase activity but no extracellular release of oxidants. The thapsigargin-induced effect was dependent on capacitative Ca2+ influx, because the effect was inhibited dose dependently by EGTA and the Ca2+ channel blocker La3+. At La3+ concentrations between 200 and 400 microM, thapsigargin also induced a massive extracellular production of superoxide anion. No other channel blockers tested induced a similar effect. We conclude that elevation in [Ca2+]i by capacitative Ca2+ influx induces NADPH-oxidase activation at an intracellular site. Further, activation of the plasma membrane-localized NADPH-oxidase is regulated by a more complex Ca2+ signaling, involving capacitative Ca2+ influx and possibly the specific action of La3+-sensitive Ca2+ channels. PMID- 11927648 TI - Nitric oxide synthase and nitric oxide production in in vivo-derived mast cells. AB - Nitric oxide (NO) is a potent mediator synthesized by a variety of cells involved in inflammatory reactions. We investigated the expression of NO synthase (NOS) in rat peritoneal mast cells (PMC). Small amounts of eNOS mRNA were detected basally, whereas neither mRNA for iNOS nor nNOS was detected in unstimulated PMC. Following stimulation by antigen, interferon-gamma (IFN-gamma), or anti-CD8 antibody, PMC up-regulated iNOS mRNA expression. In situ RT-PCR confirmed that iNOS mRNA originated from PMC. Production of iNOS protein was confirmed in stimulated PMC by immunohistochemistry. Upon stimulation with antigen, IFN-gamma, or anti-CD8, nitrite production was increased significantly (8.4+/-0.6, 7.6+/ 0.9, and 6.6+/-0.9 microM/2x10(5) cells/48 h NO2-, respectively; P<0.01), whereas unstimulated PMC released 2.1 +/- 0.3 microM/2 x 10(5) cells/48 h NO2-. These findings demonstrate that in vivo-derived PMC transcribe and translate mRNA for NOS and produce NO. PMID- 11927649 TI - Activation of macrophages by gliadin fragments: isolation and characterization of active peptide. AB - Celiac disease, induced by dietary gluten, is characterized by mucosal atrophy and local inflammation associated with cell infiltration and activation. Unlike other food proteins, gluten and its proteolytic fragments, besides inducing a specific immune response, were shown to activate components of innate immunity and cause, e.g., direct stimulation of TNF-alpha and IL-10 and a significant rise in NO production by peritoneal macrophages. The identity of the active fragments was established by separating the peptic digest of gliadin by RP-HPLC chromatography. The purest fraction with the highest activity was analyzed by mass spectrometry, and the gliadin peptide sequence was identified as VSFQQPQQQYPSSQ. This peptide (T) and its N- and C-terminally shortened forms (A, B, C and D, E, F) were synthesized. Peptide B (FQQPQQQYPSSQ) elicited the highest TNF-alpha, IL-10, and RANTES secretion and increase in IFN-gamma-primed NO production by mouse macrophages. In contrast, C-terminally shortened peptides had a lower ability to stimulate macrophages than the native form. PMID- 11927650 TI - Inhibitory actions of glucosamine, a therapeutic agent for osteoarthritis, on the functions of neutrophils. AB - Glucosamine, an amino monosaccharide naturally occurring in the connective and cartilage tissues, contributes to maintaining the strength, flexibility, and elasticity of these tissues. In recent years, glucosamine has been used widely to treat osteoarthritis in humans and animal models. Neutrophils, which usually function as the primary defenders in bacterial infections, are also implicated in the destructive, inflammatory responses in arthritis. In this study, we have evaluated the effects of glucosamine on neutrophil functions using human peripheral blood neutrophils. Glucosamine (0.01-1 mM) dose-dependently suppressed the superoxide anion generation induced by formyl-Met-Leu-Phe (fMLP) or complement-opsonized zymosan and inhibited the phagocytosis of complement opsonized zymosan or IgG-opsonized latex particles. Furthermore, glucosamine inhibited the release of granule enzyme lysozyme from phagocytosing neutrophils and suppressed neutrophil chemotaxis toward zymosan-activated serum. In addition, glucosamine inhibited fMLP-induced up-regulation of CD11b significantly, polymerization of actin, and phosphorylation of p38 mitogen-activated protein kinase (MAPK). In contrast, N-acetyl-glucosamine, an analogue of glucosamine, did not affect these neutrophil functions (superoxide generation, phagocytosis, granule enzyme release, chemotaxis, CD11b expression, actin polymerization, and p38 MAPK phosphorylation) at the concentrations examined (1-10 mM). Together these observations likely suggest that glucosamine suppresses the neutrophil functions, thereby possibly exhibiting anti-inflammatory actions in arthritis. PMID- 11927651 TI - Physiological levels of 1alpha, 25 dihydroxyvitamin D3 induce the monocytic commitment of CD34+ hematopoietic progenitors. AB - Although supraphysiological levels of 1alpha, 25 dihydroxyvitamin D3 (VD) have been demonstrated extensively to induce the monomacrophagic differentiation of leukemic myelo- and monoblasts, little is known about the role that physiological levels of this vitamin could play in the regulation of normal hematopoiesis. To clarify this issue, we adopted a liquid-culture model in which cord blood CD34+ hematopoietic progenitors, induced to differentiate in the presence of different combinations of cytokines, were exposed to VD at various concentrations and stimulation modalities. The data obtained show that physiological levels of VD promote a differentiation of CD34+ hematopoietic progenitors characterized by the induction of all the monomacrophagic immunophenotypic and morphological markers. This effect is not only exerted at the terminal maturation but also at the commitment level, as demonstrated by the decrease of highly undifferentiated CD34+CD38- hematopoietic stem cells, the down-regulation of CD34 antigen, and the increase of monocyte-committed progenitors. Molecular analysis suggests that the VD genomic signaling pathway underlies the described differentiation effects. PMID- 11927652 TI - Erythrocytes deliver Tat to interferon-gamma-treated human dendritic cells for efficient initiation of specific type 1 immune responses in vitro. AB - Dendritic cells (DC) can represent an important target for vaccine development against viral infections. Here, we studied whether interferon-gamma (IFN-gamma) could improve the functions of DC and analyzed human red blood cells (RBC) as a delivery system for Tat protein. Monocyte-derived DC were cultured in human serum and matured with monocyte-conditioned medium (MCM) in the presence or not of IFN gamma. Tat was conjugated to RBC (RBC-Tat) through avidin-biotin bridges. Stimulation of DC with IFN-gamma increased the release of interleukin (IL)-12 and tumor necrosis factor-alpha and inhibited the production of IL-10. Moreover, IFN gamma-treated DC up-regulated the release of CXCL10 (IP-10) markedly and reduced the secretion of CCL17 TARC significantly, attracting preferentially T-helper (Th)1 and Th2 cells, respectively. DC internalized RBC-Tat efficiently. Compared with DC pulsed with soluble Tat, DC incubated with RBC-Tat elicited specific CD4+ and CD8+ T-cell responses at a much lower antigen dose. DC matured in the presence of MCM were more effective than immature DC in inducing T-cell proliferation and IFN-gamma release. Finally, immature and mature DC exposed to IFN-gamma were better stimulators of allogeneic T cells and induced a higher IFN gamma production from Tat-specific CD4+ and CD8+ T lymphocytes. In conclusion, erythrocytes appear an effective tool for antigen delivery into DC, and IFN-gamma could be used advantageously for augmenting the ability of DC to induce type 1 immune responses. PMID- 11927653 TI - T cell-mediated signaling to vascular endothelium: induction of cytokines, chemokines, and tissue factor. AB - Adhesion of leukocytes to the vascular endothelium is an early event in inflammation. Since cell-cell signaling may be an important stimulus for endothelial activation, we focused in this study on the role of contact-mediated activation by T lymphocytes of endothelial cells (EC). T lymphocytes were cultured with anti-CD3 monoclonal antibody or in the presence of a combination of TNF-alpha, interleukin (IL)-6, and IL-2, prior to fixation and coculture with human umbilical vein EC. Fixed, activated (anti-CD3- or cytokine-stimulated), but not unstimulated T cells, induced release of monocyte chemotactic protein-1, IL 8, and IL-6 by EC in a contact-dependent manner. Moreover, expression of tissue factor antigen and activity was also significantly increased. Addition of anti CD40 ligand antibody abolished T cell-induced activation of EC. Our data suggest that contact-mediated activation of EC by T cells, involving ligand:counter ligand interactions such as CD40:CD40 ligand, may represent a novel pathogenic mechanism of progression in inflammatory diseases such as atherosclerosis or rheumatoid arthritis. PMID- 11927654 TI - IFN-alpha2a induces IP-10/CXCL10 and MIG/CXCL9 production in monocyte-derived dendritic cells and enhances their capacity to attract and stimulate CD8+ effector T cells. AB - Type I IFNs are immunomodulatory factors that possibly influence the properties of tissue-resident dendritic cells. Here, we have investigated the capacity of IFN-alpha2a to enhance DC chemoattractive and stimulatory capacity toward CD8+ T lymphocytes. Phenotypically, IFN-alpha2a-treated DC (IFN-DC) showed an increased expression of costimulatory and antigen-presenting molecules, maintained even after withdrawal of the cytokine. IFN-alpha2a enhanced DC stimulatory capacity toward CD8+ T cells, as assessed by increased MLR responses and induction of MART 1(26-35)-specific CTLs in vitro. No functional CCR7 chemokine receptor could be induced. Instead, high amounts of IP-10/CXCL10 and MIG/CXCL9 chemokines were produced. Freshly isolated CD8+RO+ cells and PHA-activated CD8+ T cells migrated efficiently in response to IFN-DC-conditioned medium, and the migration could be inhibited by neutralizing the CXCR3 receptor on responder cells. These results suggest that type I IFNs could enhance the elicitation of class I-restricted effector functions in vivo in the periphery by modulating DC chemoattractive properties. PMID- 11927655 TI - Regulation of murine macrophage proinflammatory and anti-inflammatory cytokines by ligands for peroxisome proliferator-activated receptor-gamma: counter regulatory activity by IFN-gamma. AB - The prostaglandin, 15-deoxy Delta(12,14)-prostaglandin J2 (15d-PGJ2)(1), and thiazolidinediones are ligands for the nuclear receptor, peroxisome proliferator activated receptor (PPAR)-gamma, which mediates anti-inflammatory activity by suppressing murine macrophage (Mphi) production of the inflammatory mediator, nitric oxide (NO). Here, we elucidated this anti-inflammatory activity further by investigating whether PPAR-gamma ligands regulated a panel of proinflammatory and anti-inflammatory cytokines produced by primary inflammatory murine Mphi (thioglycollate-elicited peritoneal exudate Mphi; PEM). Thiazolidinediones and 15d-PGJ2 suppressed lipopolysaccharide (LPS)-induced PEM production of NO and IL 12(p40) to a greater extent than IL-6 and TNF-alpha production. Whereas 15d-PGJ2 showed the greatest extent of suppression of proinflammatory mediator production, the thiazolidinedione, BRL49653, was the most potent compound studied. Surprisingly, treatment with the Mphi-activation cytokine, IFN-gamma, prevented PPAR-gamma ligands from suppressing the proinflammatory cytokines completely and reduced their suppression of NO production substantially, demonstrating that activation conditions affect PPAR-gamma-mediated, anti-inflammatory activity. Western analysis demonstrated that the antagonistic activity of IFN-gamma did not involve modulation of PPAR-gamma expression but showed that IFN-gamma interfered with PPAR-gamma ligand regulation of p42/p44 MAP kinase activation and the cytosolic disappearance of NF-kappaB upon LPS stimulation. Finally, we showed that PPAR-gamma ligands did not substantially modulate production of the anti inflammatory cytokine, IL-10, and that antibody-mediated neutralization of IL-10 did not prevent the ligands from suppressing proinflammatory mediator production. In contrast to studies with noninflammatory human monocytes and Mphi, our results demonstrate that primary murine inflammatory Mphi are extremely sensitive to the anti-inflammatory activity of PPAR-gamma ligands. These results suggest that drugs such as thiazolidinediones may be most effective in suppressing Mphi activity early (i.e., in the absence of lymphocyte-derived IFN-gamma) in the inflammatory process. PMID- 11927656 TI - Inhibition of murine macrophage nitric oxide production by synthetic oligonucleotides. AB - Synthetic 30-mer phosphorothioate (Ps) oligonucleotides (ODN) comprised of single bases (SdA30, SdC30, SdG30, and SdT30) were assessed for their effects on nitric oxide (NO) production by murine bone marrow macrophages (BMMC) and macrophage cell lines J774 and RAW264.7. Pretreatment of these cells with any of the four Ps ODN inhibited NO production induced by CpG ODN, E. coli DNA (EC DNA), or LPS. This inhibition was time- and dose-dependent and was observed even if the Ps ODN were added as long as 12 h after stimulation. As in the case of stimulatory ODN, inhibition was dependent on backbone structure and length. Thus, all four 30-mer, single-base Ps ODN were inhibitory, and only dG30 among phosphodiester ODN was inhibitory. Together, these observations indicate that Ps ODN can inhibit macrophage production of inflammatory mediators, suggesting a role of these compounds as immunomodulatory agents. PMID- 11927657 TI - Contribution of phopholipase D and a brefeldin A-sensitive ARF to chemoattractant induced superoxide production and secretion of human neutrophils. AB - We show that blockers of phospholipase D (PLD) reduce fMLP-triggered exocytosis of secretory vesicles effectively. In accordance with this, the PLD product phosphatidic acid (PA) was able to induce mobilization of secretory vesicles. Although PLD seems to play a role in the release of all neutrophil granule types, exogenous PA alone was not sufficient to activate the exocytosis of primary and secondary granules, suggesting that in the case of these granules, additional signaling factors are required to initiate the secretory responses. The ADP ribosylation factor (ARF)-inhibitor brefeldin A (BFA) inhibited the fMLP stimulated O2*- production strongly, whereas it did not influence any of the exocytic responses, and no significant effect of BFA was detected on the O2*- generation induced by other stimuli. On the basis of these results, we propose that upon chemoattractant stimulation, PLD activity is involved in induction of degranulation and O2*- production, but a BFA-sensitive ARF is only required to the activation of the NADPH oxidase. This ARF action seems to participate exclusively in the signaling pathway between the fMLP receptor and the oxidase. PMID- 11927658 TI - Signaling to localized degranulation in neutrophils adherent to immune complexes. AB - The present study demonstrates that the secretion of azurophilic granules occurring during Fc receptor-mediated attachment and spreading of neutrophils is highly localized to the adhering region of the cell. In contrast, the secretion of specific granules occurs in a nonpolarized way. This implies that unique signals are involved in the regulation of azurophilic degranulation. Assembly of actin filaments, as visualized by staining with rhodamine phalloidin, neither hindered nor facilitated degranulation. Further, the azurophilic secretory response remained localized in the presence of cytochalasin B. Release of azurophilic-granule content was inhibited by genistein and erbstatin, inhibitors of tyrosine kinases, and by GF109203X, a protein kinase C (PKC) inhibitor. We could also demonstrate a relative enrichment of syk tyrosine kinase and the PKC isoforms alpha and beta1 in adherent plasma membranes. PMID- 11927659 TI - Cytokine-mediated regulation of CXCR4 expression in human neutrophils. AB - Several lines of evidence have suggested that a CXC chemokine receptor 4 (CXCR4)/stromal cell-derived factor-1 [SDF-1; CXC chemokine ligand 12 (CXCL12)] pair is involved in baseline trafficking of leukocytes into extravascular tissues and that modulation of surface CXCR4 expression may represent an alternative mechanism for control of cell-specific biological responses to SDF-1/CXCL12. We explored the regulation of CXCR4 expression by cytokines in polymorphonuclear neutrophils (PMNs). No significant surface expression of CXCR4 in freshly isolated PMNs was detected, but expression became apparent gradually during incubation. SDF-1alpha/CXCL12 initiated Ca2+ mobilization and migratory responses in 20 h cultured PMNs. The surface CXCR4 expression was suppressed most potently by interferon-gamma (IFN-gamma). IFN-alpha, granulocyte-macrophage colony stimulating factor (GM-CSF), and G-CSF also inhibited spontaneous CXCR4 expression. Real-time, quantitative PCR experiments revealed that a spontaneous increase and an IFN-gamma-mediated decrease in surface CXCR4 paralleled changes in the CXCR4 mRNA level. These results on PMNs support the argument that the SDF 1 (CXCL12)/CXCR4 system is regulated by cell type-specific mechanisms. PMID- 11927660 TI - ARNO but not cytohesin-1 translocation is phosphatidylinositol 3-kinase-dependent in HL-60 cells. AB - Cytohesin-1 and ARNO are guanine nucleotide-exchange factors (GEFs) for ADP ribosylation factor (Arf). Here, we show that ARNO is expressed in HL-60 cells and established that granulocytic differentiation induced with Me2SO stimulated cytohesin-1 but not ARNO expression. Cytohesin-1 levels in HL-60 granulocytes were similar to those in human neutrophils. Me2SO-differentiated HL-60 cells expressed ARNO and cytohesin-1 isoforms with a diglycine and a triglycine motif in their PH domains, respectively. In vitro, ARNO diglycine and cytohesin-1 triglycine enhanced phospholipase D1 (PLD1) activation by Arf1 with near-maximal effects at 250 nM. These effects were marked particularly at low Mg2+ concentrations. PLD activation was well-correlated with GTP binding to Arf1, and cytohesin-1 was always more potent than ARNO in the PLD- and GTP-binding assays. Increasing Mg2+ concentrations reduced PLD and Arf1 activation by Arf-GEFs. fMetLeuPhe and phorbol 12-myristate 13-acetate stimulated ARNO and cytohesin-1 as well as Arf1 translocation to HL-60 cell membranes. fMetLeuPhe-mediated ARNO recruitment, but not cytohesin-1 and Arf1 translocation, was blocked by phosphatidylinositol 3-kinase inhibitors. The combined results demonstrate that cytohesin-1 triglycine participates in a major phosphatidylinositol 3-kinase independent pathway linking cell-surface receptors to Arf1 activation and translocation in human granulocytes. PMID- 11927661 TI - Synergistic action of cytokines and purified respiratory syncytial virus in nitric oxide induction. PMID- 11927662 TI - K(+) recycling and gastric acid secretion. PMID- 11927663 TI - Attenuated sympathetic vasoconstriction in contracting muscles: just say NO. PMID- 11927664 TI - Specific contribution of human T-type calcium channel isotypes (alpha(1G), alpha(1H) and alpha(1I)) to neuronal excitability. AB - In several types of neurons, firing is an intrinsic property produced by specific classes of ion channels. Low-voltage-activated T-type calcium channels (T channels), which activate with small membrane depolarizations, can generate burst firing and pacemaker activity. Here we have investigated the specific contribution to neuronal excitability of cloned human T-channel subunits. Using HEK-293 cells transiently transfected with the human alpha(1G) (Ca(V)3.1), alpha(1H) (Ca(V)3.2) and alpha(1I) (Ca(V)3.3) subunits, we describe significant differences among these isotypes in their biophysical properties, which are highlighted in action potential clamp studies. Firing activities occurring in cerebellar Purkinje neurons and in thalamocortical relay neurons used as voltage clamp waveforms revealed that alpha(1G) channels and, to a lesser extent, alpha(1H) channels produced large and transient currents, while currents related to alpha(1I) channels exhibited facilitation and produced a sustained calcium entry associated with the depolarizing after-potential interval. Using simulations of reticular and relay thalamic neuron activities, we show that alpha(1I) currents contributed to sustained electrical activities, while alpha(1G) and alpha(1H) currents generated short burst firing. Modelling experiments with the NEURON model further revealed that the alpha(1G) channel and alpha(1I) channel parameters best accounted for T-channel activities described in thalamocortical relay neurons and in reticular neurons, respectively. Altogether, the data provide evidence for a role of alpha(1I) channel in pacemaker activity and further demonstrate that each T-channel pore-forming subunit displays specific gating properties that account for its unique contribution to neuronal firing. PMID- 11927665 TI - A comparison of currents carried by HERG, with and without coexpression of MiRP1, and the native rapid delayed rectifier current. Is MiRP1 the missing link? AB - Although it has been suggested that coexpression of minK related peptide (MiRP1) is required for reconstitution of native rapid delayed-rectifier current (I(Kr)) by human ether-a-go-go related gene (HERG), currents resulting from HERG (I(HERG)) and HERG plus MiRP1 expression have not been directly compared with native I(Kr). We compared the pharmacological and selected biophysical properties of I(HERG) with and without MiRP1 coexpression in Chinese hamster ovary (CHO) cells with those of guinea-pig I(Kr) under comparable conditions. Comparisons were also made with HERG expressed in Xenopus oocytes. MiRP1 coexpression significantly accelerated I(HERG) deactivation at potentials negative to the reversal potential, but did not affect more physiologically relevant deactivation of outward I(HERG), which remained slower than that of I(Kr). MiRP1 shifted I(HERG) activation voltage dependence in the hyperpolarizing direction, whereas I(Kr) activated at voltages more positive than I(HERG). There were major discrepancies between the sensitivity to quinidine, E-4031 and dofetilide of I(HERG) in Xenopus oocytes compared to I(Kr), which were not substantially affected by coexpression with MiRP1. On the other hand, the pharmacological sensitivity of I(HERG) in CHO cells was indistinguishable from that of I(Kr) and was unaffected by MiRP1 coexpression. We conclude that the properties of I(HERG) in CHO cells are similar in many ways to those of native I(Kr) under the same recording conditions, and that the discrepancies that remain are not reduced by coexpression with MiRP1. These results suggest that the physiological role of MiRP1 may not be to act as an essential consituent of the HERG channel complex carrying native I(Kr). PMID- 11927666 TI - Multiple inhibitory effects of Au(CN)(2-) ions on cystic fibrosis transmembrane conductance regulator Cl(-) channel currents. AB - Lyotropic pseudohalide anions are potentially useful as high affinity probes of Cl(-) channel pores. However, the interaction between these pseudohalides and the cystic fibrosis transmembrane conductance regulator (CFTR) Cl(-) channel have not been described in detail. Here we show that Au(CN)(2-) ions applied to the intracellular face of membrane patches from stably transfected baby hamster kidney cells inhibit CFTR channel currents by at least two mechanisms, which can be distinguished at the single channel level or by inhibiting channel closure using 2 mM pyrophosphate. Low concentrations (< 10 microM) of Au(CN)(2-) significantly reduced CFTR channel open probability. This effect was apparently voltage insensitive, independent of extracellular Cl(-) concentration, and lost following exposure to pyrophosphate. Higher concentrations of intracellular Au(CN)(2-) caused an apparent reduction in unitary current amplitude, presumably due to a kinetically fast blocking reaction. This effect, isolated following exposure to pyrophosphate, was strongly voltage dependent (apparent K(d) 61.6 microM at -100 mV and 913 microM at +60 mV). Both the affinity and voltage dependence of block were highly sensitive to extracellular Cl(-) concentration. We propose that Au(CN)(2-) has at least two inhibitory effects on CFTR currents: a high affinity effect on channel gating due to action on a cytoplasmically accessible aspect of the channel and a lower affinity block within the open channel pore. These results offer important caveats for the use of lyotropic pseudohalide anions such as Au(CN)(2-) as specific high affinity probes of Cl(-) channel pores. PMID- 11927667 TI - Molecular determinants of Au(CN)(2)(-) binding and permeability within the cystic fibrosis transmembrane conductance regulator Cl(-) channel pore. AB - Lyotropic anions with low free energy of hydration show both high permeability and tight binding in the cystic fibrosis transmembrane conductance regulator (CFTR) Cl(-) channel pore. However, the molecular bases of anion selectivity and anion binding within the CFTR pore are not well defined and the relationship between binding and selectivity is unclear. We have studied the effects of point mutations throughout the sixth transmembrane (TM6) region of CFTR on channel block by, and permeability of, the highly lyotropic Au(CN)(2)(-) anion, using patch clamp recording from transiently transfected baby hamster kidney cells. Channel block by 100 microM Au(CN)(2)(-), a measure of intrapore anion binding affinity, was significantly weakened in the CFTR mutants K335A, F337S, T338A and I344A, significantly strengthened in S341A and R352Q and unaltered in K329A. Relative Au(CN)(2)(-) permeability was significantly increased in T338A and S341A, significantly decreased in F337S and unaffected in all other mutants studied. These results are used to define a model of the pore containing multiple anion binding sites but a more localised anion selectivity region. The central part of TM6 (F337-S341) appears to be the main determinant of both anion binding and anion selectivity. However, comparison of the effects of individual mutations on binding and selectivity suggest that these two aspects of the permeation mechanism are not strongly interdependent. PMID- 11927668 TI - Bile acids induce calcium signals in mouse pancreatic acinar cells: implications for bile-induced pancreatic pathology. AB - The effect of the natural bile acid, taurolithocholic acid 3-sulfate (TLC-S), on calcium signalling in pancreatic acinar cells has been investigated. TLC-S induced global calcium oscillations and extended calcium transients as well as calcium signals localised to the secretory granule (apical) region of acinar cells. These calcium signals could still be triggered by TLC-S in a calcium-free external solution. TLC-S-induced calcium signals were not inhibited by atropine, but were abolished by caffeine or by depletion of calcium stores, due to prolonged application of ACh. Global calcium signals, produced by TLC-S application, displayed vectorial apical-to-basal polarity. The signals originated in the apical part and were then propagated to the basal region. Other natural bile acids, taurocholate (TC) and taurodeoxycholate (TDC), were also able to produce local and global calcium oscillations (but at higher concentrations than TLC-S). Bile, which can enter pancreas by reflux, has been implicated in the pathology of acute pancreatitis. The calcium releasing properties of bile acids suggest that calcium toxicity could be an important contributing factor in the bile acid-induced cellular damage. PMID- 11927669 TI - Control of the propagation of dendritic low-threshold Ca(2+) spikes in Purkinje cells from rat cerebellar slice cultures. AB - To investigate the ionic mechanisms controlling the dendrosomatic propagation of low-threshold Ca(2+) spikes (LTS) in Purkinje cells (PCs), somatically evoked discharges of action potentials (APs) were recorded under current-clamp conditions. The whole-cell configuration of the patch-clamp method was used in PCs from rat cerebellar slice cultures. Full blockade of the P/Q-type Ca(2+) current revealed slow but transient depolarizations associated with bursts of fast Na(+) APs. These can occur as a single isolated event at the onset of current injection, or repetitively (i.e. a slow complex burst). The initial transient depolarization was identified as an LTS Blockade of P/Q-type Ca(2+) channels increased the likelihood of recording Ca(2+) spikes at the soma by promoting dendrosomatic propagation. Slow rhythmic depolarizations shared several properties with the LTS (kinetics, activation/inactivation, calcium dependency and dendritic origin), suggesting that they correspond to repetitively activated dendritic LTS, which reach the soma when P/Q channels are blocked. Somatic LTS and slow complex burst activity were also induced by K(+) channel blockers such as TEA (2.5 x 10(-4) M) charybdotoxin (CTX, 10(-5) M), rIberiotoxin (10(-7) M), and 4-aminopyridine (4-AP, 10(-3) M), but not by apamin (10(-4) M). In the presence of 4-AP, slow complex burst activity occurred even at hyperpolarized potentials (-80 mV). In conclusion, we suggest that the propagation of dendritic LTS is controlled directly by 4-AP-sensitive K(+) channels, and indirectly modulated by activation of calcium-activated K(+) (BK) channels via P/Q-mediated Ca(2+) entry. The slow complex burst resembles strikingly the complex spike elicited by climbing fibre stimulation, and we therefore propose, as a hypothesis, that dendrosomatic propagation of the LTS could underlie the complex spike. PMID- 11927670 TI - Subplasmalemmal endoplasmic reticulum controls K(Ca) channel activity upon stimulation with a moderate histamine concentration in a human umbilical vein endothelial cell line. AB - This study was designed to elucidate the role of the subplasmalemmal endoplasmic reticulum (sER) in autacoid-induced stimulation of Ca(2+)-dependent K(+) channels in the umbilical vein endothelial cell-derived cell line EA.hy926. Cells were transfected with the Ca(2+) probe cameleon targeted to the ER for visualization of the ER network. A patch pipette was then placed close to or far (> 5 microm away) from the sER, single channel recordings (patch clamp technique) were monitored simultaneously with measurements of either ER Ca(2+) concentration (using the Ca(2+) probe Cam4-ER) or cytosolic free Ca(2+) concentration ([Ca(2+)](i); using fura-2) using a deconvolution imaging device. A voltage dependent, large conductance Ca(2+)-dependent K(+) channel (BK(Ca); single channel conductance (gamma), 250 pS) was found. At membrane potentials of +40 and -40 mV, the EC(50) for Ca(2+) was 2.7 and 49.7 microM, respectively. In the vicinity of the sER, the BK(Ca) channel activity induced by 10 microM histamine was 32 times higher (open probability (P(o)) = 0.083 +/- 0.026) than in areas away from the sER (P(o) = 0.0026 +/- 0.002). However, at supramaximal histamine stimulation (100 microM), BK(Ca) channel activation was similar in patches in the vicinity of or away from the sER (P(o) = 0.18 +/- 0.09 and 0.25 +/- 0.07, respectively). In contrast to BK(Ca) channel activity, ER Ca(2+) depletion (Cam4 ER) and elevation of [Ca(2+)](i) in response to 10 and 100 microM histamine were not influenced by the pipette position. We conclude that in endothelial cells, the activation of BK(Ca) channels in response to moderate histamine concentration essentially depends on the proximity of the sER domains to the mouth of this K(+) channel. These findings further support our concept of the subplasmalemmal Ca(2+) control unit (SCCU) and add the local activation of Ca(2+)-activated K(+) channels to the function of the SCCU. PMID- 11927671 TI - Specific localization of an inwardly rectifying K(+) channel, Kir4.1, at the apical membrane of rat gastric parietal cells; its possible involvement in K(+) recycling for the H(+)-K(+)-pump. AB - Hydrochloric acid (HCl) is produced in parietal cells of gastric epithelium by a H(+)-K(+) pump. Protons are secreted into the gastric lumen in exchange for K(+) by the action of the H(+)-K(+)-ATPase. Luminal K(+) is essential for the operation of the pump and is thought to be supplied by unidentified K(+) channels localized at the apical membrane of parietal cells. In this study, we showed that histamine- and carbachol-induced acid secretion from isolated parietal cells monitored by intracellular accumulation of aminopyrine was depressed by Ba(2+), an inhibitor of inwardly rectifying K(+) channels. Among members of the inwardly rectifying K(+) channel family, we found with reverse transcriptase-polymerase chain reaction analyses that Kir4.1, Kir4.2 and Kir7.1 were expressed in rat gastric mucosa. With immunohistochemical analyses, Kir4.1 was found to be expressed in gastric parietal cells and localized specifically at their apical membrane. The current flowing through Kir4.1 channel expressed in HEK293T cells was not affected by reduction of extracellular pH from 7.4 to 3. These results suggest that Kir4.1 may be involved in the K(+) recycling pathway in the apical membrane which is required for activation of the H(+)-K(+) pump in gastric parietal cells. PMID- 11927672 TI - Sodium current modulation by a tubulin/GTP coupled process in rat neonatal cardiac myocytes. AB - Microtubule disassembly by colchicine increases spontaneous beating of neonatal cardiac myocytes by an unknown mechanism. Here, we measure drug effects on spontaneous calcium transients and whole cell ionic currents to define the route between microtubule depolymerization and the increase in the rate of contraction. Colchicine treatment disassembles microtubules resulting in free tubulin dimers, thereby increasing the spontaneous beating frequency and changing both the rates of rise and decay of calcium transients. In addition, colchicine treatment produces an increase of the sodium current (I(Na)) while I(Ca) is not modified. The colchicine-enhanced I(Na) was blocked by the addition of 10 microM TTX. In addition, the colchicine-induced increase of I(Na) was prevented when GTP was omitted from the patch pipette. Vinblastine also depolymerizes microtubules but re-aggregates tubulin into paracrystalline structures. Free tubulin dimers are not increased with vinblastine treatment. We found no modification in calcium transients or I(Na) in the presence of vinblastine. Action potential durations measured at 50 % and 90 % repolarization were shorter, and the dV/dt was larger, in colchicine-treated cells compared to untreated cells. The resting membrane potential and overshoot of the action potentials were comparable in both kinds of cells. Our data suggest that release of free tubulin dimers may activate G proteins, which in turn modulate the sodium channel. An increase in whole cell I(Na) changes the spontaneous firing rate and this may be the underlying cause of the increase in the frequency of contraction in neonatal cardiac myocytes. We suggest a new role for dimeric tubulin in regulating membrane excitability. PMID- 11927674 TI - Dopamine selectively reduces GABA(B) transmission onto dopaminergic neurones by an unconventional presynaptic action. AB - The functioning of midbrain dopaminergic neurones is closely involved in mental processes and movement. In particular the modulation of the inhibitory inputs on these cells might be crucial in controlling firing activity and dopamine (DA) release in the brain. Here, we report a concentration-dependent depressant action of dopamine on the GABA(B) IPSPs intracellularly recorded from dopaminergic neurones. Such effect was observed in spite of the presence of D(1)/D(2) dopamine receptor antagonists. A reduction of the GABA(B) IPSPs was also caused by noradrenaline (norepinephrine) and by L-beta-3,4-dihydroxyphenylalanine (L-DOPA), which is metabolically transformed into DA. The DA-induced depression of the IPSPs was partially antagonised by the alpha2 antagonists yohimbine and phentolamine. DA did not change the postsynaptic effects of the GABA(B) agonist baclofen, suggesting a presynaptic site of action. Furthermore, DA did not modulate the GABA(A)-mediated IPSP. The DA-induced depression of the GABA(B) IPSP occluded the depression produced by serotonin and was not antagonized by serotonin antagonists. The DA- and 5-HT-induced depression of the GABA(B) IPSP persisted when calcium and potassium currents were reduced in to the presynaptic terminals. These results describe an unconventional presynaptic, D(1) and D(2) independent action of DA on the GABA(B) IPSP. This might have a principal role in determining therapeutic/side effects of L-DOPA and antipsychotics and could be also involved in drug abuse. PMID- 11927673 TI - Ca(2+)-activated Cl(-) current in rabbit sinoatrial node cells. AB - The Ca(2+)-activated Cl(-) current (I(Cl(Ca))) has been identified in atrial, Purkinje and ventricular cells, where it plays a substantial role in phase-1 repolarization and delayed after-depolarizations. In sinoatrial (SA) node cells, however, the presence and functional role of I(Cl(Ca)) is unknown. In the present study we address this issue using perforated patch-clamp methodology and computer simulations. Single SA node cells were enzymatically isolated from rabbit hearts. I(Cl(Ca)) was measured, using the perforated patch-clamp technique, as the current sensitive to the anion blocker 4,4'-diisothiocyanostilbene-2,2' disulphonic acid (DIDS). Voltage clamp experiments demonstrate the presence of I(Cl(Ca)) in one third of the spontaneously active SA node cells. The current was transient outward with a bell-shaped current-voltage relationship. Adrenoceptor stimulation with 1 microM noradrenaline doubled the I(Cl(Ca)) density. Action potential clamp measurements demonstrate that I(Cl(Ca)) is activate late during the action potential upstroke. Current clamp experiments show, both in the absence and presence of 1 microM noradrenaline, that blockade of I(Cl(Ca)) increases the action potential overshoot and duration, measured at 20 % repolarization. However, intrinsic interbeat interval, upstroke velocity, diastolic depolarization rate and the action potential duration measured at 50 and 90 % repolarization were not affected. Our experimental data are supported by computer simulations, which additionally demonstrate that I(Cl(Ca)) has a limited role in pacemaker synchronization or action potential conduction. In conclusion, I(Cl(Ca)) is present in one third of SA node cells and is activated during the pacemaker cycle. However, I(Cl(Ca)) does not modulate intrinsic interbeat interval, pacemaker synchronization or action potential conduction. PMID- 11927675 TI - Effects of daily spontaneous running on the electrophysiological properties of hindlimb motoneurones in rats. AB - No evidence currently exists that motoneurone adaptations in electrophysiological properties can result from changes in the chronic level of neuromuscular activity. We examined, in anaesthetized (ketamine/xylazine) rats, the properties of motoneurones with axons in the tibial nerve, from rats performing daily spontaneous running exercise for 12 weeks in exercise wheels ('runners') and from rats confined to plastic cages ('controls'). Motoneurones innervating the hindlimb via the tibial nerve were impaled with sharp glass microelectrodes, and the properties of resting membrane potential, spike threshold, rheobase, input resistance, and the amplitude and time-course of the afterhyperpolarization (AHP) were measured. AHP half-decay time was used to separate motoneurones into 'fast' (AHP half-decay time < 20 ms) and 'slow' (AHP half-decay time >/= 20 ms), the proportions of which were not significantly different between controls (58 % fast) and runners (65 % fast). Two-way ANOVA and ANCOVA revealed differences between motoneurones of runners and controls which were confined to the 'slow' motoneurones. Specifically, runners had slow motoneurones with more negative resting membrane potentials and spike thresholds, larger rheobasic spike amplitudes, and larger amplitude AHPs compared to slow motoneurones of controls. These adaptations were not evident in comparing fast motoneurones from runners and controls. This is the first demonstration that physiological modifications in neuromuscular activity can influence basic motoneurone biophysical properties. The results suggest that adaptations occur in the density, localization, and/or modulation of ionic membrane channels that control these properties. These changes might help offset the depolarization of spike threshold that occurs during rhythmic firing. PMID- 11927676 TI - Contractile responses of smooth muscle cells differentiated from rat neural stem cells. AB - To characterize the functional differentiation of neural stem cells into smooth muscle cells, multipotent stem cells in the central nervous system (CNS) were isolated from rat embryonic day 14 (E14) cortex and cultured by neurosphere formation in serum-free medium in the presence of 10 ng ml(-1) of basic fibroblast growth factor. Differentiation was induced by the addition of 10 % fetal bovine serum to low-density cultures (2.5 x 10(3) cells cm(-2)). Immunological analyses and reverse transcriptase-polymerase chain reaction indicated that the differentiated cells expressed smooth-muscle-specific marker proteins such as SM-1, SM-2, and SMemb myosin heavy chains, SM-22, basic calponin and alpha-smooth-muscle actin, but not the astrocyte marker glial fibrillary acidic protein. To examine whether smooth-muscle-like cells that are differentiated from CNS stem cells possess the characteristics of contractile smooth muscle, we prepared reconstituted collagen gel fibres and measured their contractile tension. The reconstituted fibres were prepared by thermal gelation of collagen and the differentiated cells. The fibres contracted in response to treatment with KCl (80 mM), ACh (100 microM), endothelin-1 (10 nM), endothelin-2 (10 nM), and prostaglandin F2alpha (100 microM). ACh-induced contraction was partially inhibited by the L-type voltage-dependent Ca(2+) channel inhibitor nifedipine and by the intracellular Ca(2+) chelator 1,2-bis (2-aminophenoxy) ethane-N,N,N',N'-tetraacetic acid acetoxymethyl ester, the myosin light chain kinase inhibitor ML-9, the Rho kinase inhibitor Y-27632, dibutyryl cAMP and 8 bromo-cGMP. These results suggest that CNS stem cells give rise to smooth muscle cells in vitro that have an identical contractile function to smooth muscle in vivo. PMID- 11927677 TI - Cleavage-arrested cell triplets from ascidian embryo differentiate into three cell types depending on cell combination and contact timing. AB - During early ascidian development, which is a prototype of early vertebrate development, anterior neuroectoderm cells (a4.2) from the eight-cell embryo are destined to become anterior neural structures including the brain vesicle, while presumptive notochordal neural cells (A4.1) become larval posterior neural structures including motoneurons. Whereas, an anterior quadrant cell (A3) of the four-cell embryo, from which both anterior neuroectoderm (a4.2) and notochordal neural cells (A4.1) are derived, has both fates. Cleavage-arrested cell triplets were prepared from the anterior quadrant cell and a pair of anterior neuroectoderm cells (A3-aa triplet) or a pair of presumptive notochordal neural cells (A3-AA triplet), and cultured in contact. Differentiation of cells in the triplet was determined electrophysiologically by observing cell type-specific currents. In the A3-aa triplet, when two neuroectoderm cells and an anterior quadrant cell were prepared from the same batch of embryos, all three cells in the triplet developed into neuronal cells in 60 % of cases, but in 40 % of cases all of them differentiated into epidermal cells. However, when the batch of embryos from which neuroectoderm cells were prepared was fertilized 3 h later than that from which the anterior quadrant cell was prepared all three cells in the triplet consistently became neuronal cells. In contrast, when the batch of embryos from which neuroectoderm cells were prepared was fertilized 3 h earlier, all three cells became epidermal. In the A3-AA triplet no switching of differentiation occurred and all three cells in the triplet differentiated into neuronal cells, although the amplitude of inward current was often small. In neuralized A3-aa triplets the spikes in the anterior quadrant cell were characteristically small in amplitude and brief in duration, suggesting the presence of A-currents, which is a characteristic feature of posterior neuronal differentiation. In contrast, the spikes in the anterior neuroectoderm cells were large in amplitude and long in duration, chracteristic to the anterior neuronal type. The majority of single isolated anterior quadrant cells became non excitable. However, the minority was apparently autonomously neuralized to become the posterior neuronal type. In neuralized A3-AA triplets, the majority of anterior quadrant cells was induced to become the anterior neuronal type. When isolated anterior quadrant cells were neuralized with subtilisin, a protease, they also predominantly became the anterior neuronal type. While, in medium containing a fibroblast growth factor posterior neuralization of isolated anterior quadrant cells was facilitated, but the anterior neuronal type, although minor, appeared anew. These observations indicate that the multiple fates of the anterior quadrant cell expressed in vivo were effectively reproduced in this experimental condition at the single cell level. Interactive differentiation in this triplet system recapitulates not only fundamental neural induction of ascidian neuroectoderm cells, but also functional and positional specificity within the neuronal group. PMID- 11927678 TI - Titin-based contribution to shortening velocity of rabbit skeletal myofibrils. AB - The shortening velocity of skeletal muscle fibres is determined principally by actomyosin cross-bridges. However, these contractile elements are in parallel with elastic elements, whose main structural basis is thought to be the titin filaments. If titin is stretched, it may contribute to sarcomere shortening simply because it can recoil 'passively'. The titin-based contribution to shortening velocity (V(p)) was quantified in single rabbit psoas myofibrils. Non activated specimens were rapidly released from different initial sarcomere lengths (SLs) by various step amplitudes sufficient to buckle the myofibrils; V(p) was calculated from the release amplitude and the time to slack reuptake. V(p) increased progressively (upper limit of detection, approximately 60 microm s(-1) sarcomere(-1)) between 2.0 and 3.0 microm SL, albeit more steeply than passive tension. At very low passive tension levels already (< 1-2 mN mm(-2)), V(p) could greatly exceed the unloaded shortening velocity measured in fully Ca(2+)-activated skinned rabbit psoas fibres. Degradation of titin in relaxed myofibrils by low doses of trypsin (5 min) drastically decreased V(p). In intact myofibrils, average V(p) was faster, the smaller the release step applied. Also, V(p) was much higher at 30 degrees C than at 15 degrees C (Q(10): 2.0, 3.04 or 6.15, for release steps of 150, 250 or 450 nm sarcomere(-1), respectively). Viscous forces opposing the shortening are likely to be involved in determining these effects. The results support the idea that the contractile system imposes a braking force onto the passive recoil of elastic structures. However, elastic recoil may aid active shortening during phases of high elastic energy utilization, i.e. immediately after the onset of contraction under low or zero load or during prolonged shortening from greater physiological SLs. PMID- 11927679 TI - Correlations between neuronal morphology and electrophysiological features in the rodent superficial dorsal horn. AB - Relationships between the morphology of individual neurones of the spinal superficial dorsal horn (SDH), laminae I and II, and their electrophysiological properties were studied in spinal cord slices prepared from anaesthetized, free ranging hamsters. Tight-seal, whole-cell recordings were made with pipette microelectrodes filled with biocytin to establish electrophysiological characteristics and to label the studied neurones. Neurones were categorized according to location and size of the somata, the dendritic and axonal pattern of arborization, spontaneous synaptic potentials, evoked postsynaptic currents, pattern of discharge to depolarizing pulses and current-voltage relationships. Data were obtained for 170 neurones; 13 of these had somata in lamina I and 157 in lamina II. Stimulation of the segmental dorsal root evoked a prompt excitatory response in almost every neurone sampled (161/166) with nearly 3/4 displaying putative monosynaptic EPSCs. The majority of neurones (133/170) fitted one of several distinctive morphological categories. To a considerable extent, neurones with a common morphological configuration and neurite disposition shared electrophysiological characteristics. Five of the 13 lamina I neurones were relatively large with extensive dendritic arborization in the horizontal dimension and a prominent axon directed ventrally and contralaterally. These presumptive ventrolateral projection neurones differed structurally and electrophysiologically from the other lamina I neurones, which had ipsilateral, locally arborizing axons and/or branches entering the dorsal lateral funiculus. One hundred and twenty lamina II neurones fitted one of five morphological categories: islet, central, medial-lateral, radial or vertical. Central cells were further divided into three groups on functional features. We conclude that the spinal SDH comprises many types of neurones whose morphological characteristics are associated with specific functional features implying diversity in functional organization of the SDH and in its role as a major synaptic termination for thin primary afferent fibres. PMID- 11927680 TI - The role of guanylyl cyclases in the permeability response to inflammatory mediators in pial venular capillaries in the rat. AB - Inflammatory mediators have a role in the formation of cerebral oedema and there is evidence that cGMP is an important signal in vascular permeability increase. We have investigated the role and the source of cGMP in mediating the permeability response to acutely applied bradykinin and the histamine H(2) agonist dimaprit on single cerebral venular capillaries, by using the single vessel occlusion technique. We found that 8-bromo-cGMP applied acutely resulted in a small and reversible permeability increase with a log EC(50) -7.2 +/- 0.15 M. KT 5823, the inhibitor of cGMP-dependent protein kinase, abolished the permeability responses to both bradykinin and dimaprit, while zaprinast, an inhibitor of type 5 phosphodiesterase, potentiated the response to bradykinin. On the other hand, L-NMMA blocked the response to dimaprit, but not that to bradykinin. Inhibitors of soluble guanylyl cyclase, LY 85353 and methylene blue, also inhibited the permeability response to dimaprit, but not bradykinin. The permeability responses to the natriuretic peptides ANP and CNP were of similar magnitude to that of bradykinin with log EC(50) -10.0 +/- 0.33 M and -8.7 +/- 0.23 M, respectively. The natriuretic peptide receptor antagonist HS-142-1 blocked permeability responses to bradykinin as well as to ANP, and leukotriene D(4) blocked the responses to CNP and bradykinin, but not to dimaprit. In conclusion, the histamine H(2) receptor appears to signal via cGMP that is generated by a NO and soluble guanylyl cyclase, while bradykinin B(2) receptor also signals via cGMP but through particulate guanylyl cyclase. PMID- 11927682 TI - Coupling between the ribs and the lung in dogs. AB - In contrast to the conventional theory, the external and internal intercostal muscles show marked rostrocaudal gradients in their actions on the lung. We hypothesized that these gradients are the result of a non-uniform coupling between the ribs and the lung. Rib displacements (X(r)) and the changes in airway opening pressure (P(a,o)) were thus measured in anaesthetized, pancuronium treated, supine dogs while loads were applied in the cranial direction to individual pairs of odd-numbered ribs and in the caudal direction to individual pairs of even-numbered ribs. During cranial loading, X(r) induced by a given load increased gradually with increasing rib number. The decrease in P(a,o) also increased from the third to the fifth rib pair but then decreased markedly to the eleventh pair. A similar pattern was observed during caudal loading, although X(r) and DeltaP(a,o) were smaller. These results were then combined to calculate the net X(r) and the net DeltaP(a,o) that a hypothetical intercostal muscle lying parallel to the longitudinal body axis would produce in different interspaces. The net X(r) was cranial in all interspaces. However, whereas the net DeltaP(a,o) was negative in the cranial interspaces, it was positive in the caudal interspaces. These observations confirm that the coupling between the ribs and the lung varies from the top to the base of the ribcage. This coupling confers to both the external and the internal intercostal muscles an inspiratory action on the lung in the cranial interspaces and an expiratory action in the caudal interspaces. PMID- 11927683 TI - Sympathetic modulation of muscle spindle afferent sensitivity to stretch in rabbit jaw closing muscles. AB - Previous reports showed that sympathetic stimulation affects the activity of muscle spindle afferents (MSAs). The aim of the present work is to study the characteristics of sympathetic modulation of MSA response to stretch: (i) on the dynamic and static components of the stretch response, and (ii) on group Ia and II MSAs to evaluate potentially different effects. In anaesthetised rabbits, the peripheral stump of the cervical sympathetic nerve (CSN) was stimulated at 10 impulses s(-1) for 45-90 s. The responses of single MSAs to trapezoidal displacement of the mandible were recorded from the mesencephalic trigeminal nucleus. The following characteristic parameters were determined from averaged trapezoidal responses: initial frequency (IF), peak frequency at the end of the ramp (PF), and static index (SI). From these, other parameters were derived: dynamic index (DI = PF - SI), dynamic difference (DD = PF - IF) and static difference (SD = SI - IF). The effects of CSN stimulation were also evaluated during changes in the state of intrafusal muscle fibre contraction induced by succinylcholine and curare. In a population of 124 MSAs, 106 units (85.4 %) were affected by sympathetic stimulation. In general, while changes in resting discharge varied among different units (Ia vs. II) and experimental conditions (curarised vs. non-curarised), ranging from enhancement to strong depression of firing, the amplitude of the response to muscle stretches consistently decreased. This was confirmed and detailed in a quantitative analysis performed on 49 muscle spindle afferents. In both the non-curarised (23 units) and curarised (26 units) condition, stimulation of the CSN reduced the response amplitude in terms of DD and SD, but hardly affected DI. The effects were equally present in both Ia and II units; they were shown to be independent from gamma drive and intrafusal muscle tone and not secondary to muscle hypoxia. Sympathetic action on the resting discharge (IF) was less consistent. In the non-curarised condition, IF decreased in most Ia units, while in II units decreases and increases occurred equally often. In the curarised condition, IF in group II units mostly increased. The results have important functional implications on the control of motor function in a state of 'high' sympathetic activity, like excessive stress, as well as in certain pathological conditions such as sympathetically maintained pain. PMID- 11927681 TI - Voltage independence of vasomotion in isolated irideal arterioles of the rat. AB - The cellular mechanisms underlying vasomotion of irideal arterioles from juvenile rats have been studied using electrophysiological methods, ratiometric calcium measurements and video microscopy. Vasomotion was not affected by removal of the endothelium. Spontaneous contractions were preceded by spontaneous depolarizations. Both were abolished by the intracellular calcium chelator, BAPTA AM (20 microM), but not by ryanodine (10 microM), suggesting a dependence on the cyclical release of calcium from intracellular stores, other than those operated by ryanodine receptors. Oscillations were little changed when the membrane potential of short segments of arteriole was either depolarized or hyperpolarized. When the segments were voltage clamped, oscillating inward currents were recorded, indicating that the changes in membrane potential were voltage independent. Vasomotion was preceded by intracellular calcium oscillations and both were abolished by inhibitors of phospholipase C (U73122, 10 microM), phospholipase A(2) (AACOCF(3), 30 microM) and protein kinase C (chelerythrine chloride, 5 microM, and myristoylated protein kinase C peptide, 10 microM). Inhibition of vasomotion by the dual lipoxygenase and cyclo-oxygenase inhibitor, NDGA (10 microM), the lipoxygenase inhibitor, ETI (1 microM) but not by the cyclo-oxygenase inhibitors, aspirin (10 microM) and indomethacin (10 microM), or the cytochrome P450 inhibitor 17-ODYA (10 microM), suggested an involvement of the lipoxygenase pathway. The observations suggest that vasomotion of iris arterioles is voltage independent and results from the cyclical release of calcium from IP(3)-sensitive stores which are activated by cross talk between the phospholipase C and phospholipase A(2) pathways in vascular smooth muscle. PMID- 11927685 TI - Vasodilatation in response to repeated anodal current application in the human skin relies on aspirin-sensitive mechanisms. AB - The vasodilatation resulting from prolonged square-wave monopolar current application as used in iontophoresis is assumed to rely on an axon reflex. Involvement of prostaglandins in the anodal current-induced vasodilatation remains unclear. We tested the hypothesis that prostaglandins participate in a sensitisation mechanism to current application rather than as direct vasodilators. In healthy volunteers, laser Doppler flowmetry (LDF) was recorded in the forearm during and following isolated or repeated 0.1 mA transcutaneous anodal current applications, using deionised water as a vehicle. Segmented current applications of 6 or 12 mC resulted in an LDF increase twice that observed following current applications of comparable total charge delivered all at once (P < 0.05). Following a 1 min anodal application, a slow and prolonged LDF drift occurred (slope: 0.3 +/- 0.5 arbitrary units min(-1)). When the same current application was repeated after intervals of 5 and 20 min, an abrupt vasodilatation occurred, with maximal LDF amplitude of 53.5 +/- 34.0 and 48.2 +/- 19.1 arbitrary units, respectively. Pretreatment with 1 g oral aspirin abolished the abrupt vasodilatation to repeated current application but not the initial slow drift. We suggest that vasodilatation occurs through two parallel pathways: (1) a slow progressive drift of LDF of limited amplitude insensitive to aspirin pretreatment, and (2) an abrupt vasodilatation probably resulting from afferent fibre activation, appearing if a preliminary sensitisation by current application is performed. Sensitisation lasts for at least 20 min, and is blocked by aspirin, suggesting participation of prostanoids. PMID- 11927684 TI - Spontaneous electrical activity and associated changes in calcium concentration in guinea-pig gastric smooth muscle. AB - Spontaneous electrical activity and internal Ca(2+) concentration ([Ca(2+)](i)) were measured simultaneously using conventional microelectrodes and fura-2 fluorescence, respectively, in isolated circular smooth muscle bundles of the guinea-pig gastric antrum. The smooth muscle bundles generated periodic slow potentials with accompanying spike potentials and associated transient increases in [Ca(2+)](i) (Ca(2+)-transients). Nifedipine abolished the spike potentials but not the slow potentials, and reduced the amplitude of associated Ca(2+) transients. Caffeine, in the absence or presence of ryanodine, reduced resting [Ca(2+)](i) levels and abolished the slow potentials and associated Ca(2+) transients. Depolarization elevated and hyperpolarization reduced resting [Ca(2+)](i) levels with associated changes in the frequency of slow potentials. The amplitude of Ca(2+)-transients changed in a bell-shaped manner with the membrane potential change. Slow potentials and associated Ca(2+)-transients were abolished if [Ca(2+)](i) levels were reduced by BAPTA-AM or if the internal Ca(2+) pump was inhibited by cyclopiazonic acid. 2-Aminoethoxy-diphenylborate (2 APB), a known inhibitor of inositol trisphosphate (IP(3))-mediated Ca(2+) release, also blocked slow potentials and Ca(2+)-transients. Carbonyl cyanide m chlorophenyl hydrazone (CCCP), a mitochondrial protonophore, depolarized the membrane, elevated [Ca(2+)](i) levels and abolished slow potentials and Ca(2+) transients. Inhibition of mitochondrial ATP-sensitive K(+) channels by glybenclamide and 5-hydroxydecanoic acid (5-HAD) abolished slow potentials and Ca(2+)-transients, without altering the smooth muscle [Ca(2+)](i). It is concluded that in antrum circular muscles, the frequency of slow potentials is correlated with the level of [Ca(2+)](i). The slow potential is coupled to release of Ca(2+) from an internal store, possibly through the activation of IP(3) receptors; this may be initiated by the activation of ATP-sensitive K(+) channels in mitochondria following Ca(2+) handling by mitochondria. PMID- 11927686 TI - Interactive effect of chondroitin sulphate C and hyaluronan on fluid movement across rabbit synovium. AB - The polysaccharide hyaluronan (HA) conserves synovial fluid by keeping outflow low and almost constant over a wide pressure range ('buffering'), but only at concentrations associated with polymer domain overlap. We therefore tested whether polymer interactions can cause buffering, using HA-chondroitin sulphate C (CSC) mixtures. Also, since it has been found that capillary filtration is insensitive to the Starling force interstitial osmotic pressure in frog mesenteries, this was assessed in synovium. Hyaluronan at non-buffering concentrations (0.50-0.75 mg ml(-1)) and/or 25 mg ml(-1) CSC (osmotic pressure 68 cmH(2)O) was infused into knees of anaesthetised rabbits in vivo. Viscometry and chromatography confirmed that HA interacts with CSC. Pressure (P(j)) versus trans synovial flow (;Q(s)) relations were measured.;Q(s) was outwards for HA alone (1.2 +/- 0.9 microl min(-1) at 3 cmH(2)O, mean +/- S.E.M.; n = 6). CSC diffused into synovium and changed;Q(s) to filtration at low P(j) (-4.1 microl min(-1), 3 cmH(2)O, n = 5, P < 0.02, t test). Filtration ceased upon circulatory arrest (n = 3). At higher P(j), 0.75 mg ml(-1) HA plus CSC buffered;Q(s) to approximately 3 microl min(-1) over a wide range of P(j), with an outflow increase of only 0.04 +/- 0.02 microl min(-1) cmH(2)O(-1) (n = 4). With HA or CSC alone, buffering was absent (slopes 0.57 +/- 0.04 microl min(-1) cmH(2)O(-1) (n = 4) and 0.86 +/- 0.05 microl min(-1) cmH(2)O(-1) (n = 5), respectively). Therefore, polymer interactions can cause outflow buffering in joints. Also, interstitial osmotic pressure promoted filtration in fenestrated synovial capillaries, so the results for frog mesentery capillaries cannot be generalised. The difference is attributed to differences in pore ultrastructure. PMID- 11927687 TI - Reversible impairment of endothelium-dependent relaxation in golden hamster carotid arteries during hibernation. AB - The effects of hibernation on endothelium-dependent vasodilatation were investigated in the golden hamster carotid artery, paying special attention to hibernating body temperature (10 degrees C). To record mechanical and electrical membrane responses, we applied pharmacological (organ bath) and electrophysiological (microelectrode) techniques, using acetylcholine (ACh; 0.001 100 microM) and ATP (0.01-1000 microM) for endothelium-dependent vasodilatation and sodium nitroprusside (SNP; 0.05-10 microM) for endothelium-independent vasodilatation. At 34 degrees C, ACh, ATP and SNP each induced a relaxation or a hyperpolarization, and these responses were similar in all the preparations from control and hibernated animals. At 10 degrees C, on the other hand, ACh-induced relaxations and hyperpolarizations were reduced to approximately 35 % and 50 % of the euthermic level in controls and 1 % and 4 % of the euthermic level in hibernated animals, respectively. In contrast, at 10 degrees C, ATP induced only a contraction or depolarization in all preparations with no significant difference between control and hibernated animals. SNP-induced relaxations and hyperpolarizations obtained at 34 degrees C were not attenuated by cooling to 10 degrees C. In the presence of a P2X receptor blocker, pyridoxal phosphate-6 azophenyl-2',4'-disulphonic acid (PPADS; 5 microM), at 34 degrees C ATP-induced relaxations and hyperpolarizations were significantly enhanced whereas no responses were induced by ATP at 10 degrees C. After endothelium removal, on the other hand, ATP induced only a contraction or depolarization at both 34 degrees C and 10 degrees C. These results suggest that depression of endothelium-dependent vasodilator responses to ACh and ATP may occur in the hibernating golden hamster carotid artery. PMID- 11927688 TI - Nitric oxide facilitates GABAergic neurotransmission in the cat oculomotor system: a physiological mechanism in eye movement control. AB - Nitric oxide (NO) synthesis by prepositus hypoglossi (PH) neurons is necessary for the normal performance of horizontal eye movements. We have previously shown that unilateral injections of NO synthase (NOS) inhibitors into the PH nucleus of alert cats produce velocity imbalance without alteration of the eye position control, both during spontaneous eye movements and the vestibulo-ocular reflex (VOR). This NO effect is exerted on the dorsal PH neuropil, whose fibres increase their cGMP content when stimulated by NO. In an attempt to determine whether NO acts by modulation of a specific neurotransmission system, we have now compared the oculomotor effects of NOS inhibition with those produced by local blockade of glutamatergic, GABAergic or glycinergic receptors in the PH nucleus of alert cats. Both glutamatergic antagonists used, 2-amino-5-phosphonovaleric acid (APV) and 2,3-dihydro-6-nitro-7-sulphamoyl-benzo quinoxaline (NBQX), induced a nystagmus contralateral to that observed upon NOS inhibition, and caused exponential eye position drift. In contrast, bicuculline and strychnine induced eye velocity alterations similar to those produced by NOS inhibitors, suggesting that NO oculomotor effects were due to facilitation of some inhibitory input to the PH nucleus. To investigate the anatomical location of the putative NO target neurons, the retrograde tracer Fast Blue was injected in one PH nucleus, and the brainstem sections containing Fast Blue-positive neurons were stained with double immunohistochemistry for NO-sensitive cGMP and glutamic acid decarboxylase. GABAergic neurons projecting to the PH nucleus and containing NO-sensitive cGMP were found almost exclusively in the ipsilateral medial vestibular nucleus and marginal zone. The results suggest that the nitrergic PH neurons control their own firing rate by a NO-mediated facilitation of GABAergic afferents from the ipsilateral medial vestibular nucleus. This self-control mechanism could play an important role in the maintenance of the vestibular balance necessary to generate a stable and adequate eye position signal. PMID- 11927689 TI - A novel mechanism of response selectivity of neurons in cat visual cortex. AB - The spiking of cortical neurons critically depends on properties of the afferent stimuli. In the visual cortex, neurons respond selectively to the orientation and direction of movement of an object. The orientation and direction selectivity is improved upon transformation of the membrane potential changes into trains of action potentials. To address the question of whether the transformation of the membrane potential changes into spiking of a cell depends on the stimulus orientation and the direction of movement, we made intracellular recordings from the cat visual cortex in vivo during presentation of moving gratings of different orientations. We found that the relationship between the membrane polarization and the firing rate (input-output transfer function) depended on the stimulus orientation. The input-output transfer function was steepest during responses to the optimal stimulus; membrane depolarization of a given amplitude led to generation of more action potentials when evoked by an optimal stimulus than during non-optimal stimulation. The threshold for the action potential generation did not depend on stimulus orientation, and thus could not account for the observed difference in the transfer function. Oscillations of the membrane potential in the gamma-frequency range (25-70 Hz) were most pronounced during optimal stimulation and their strength changed in parallel with the changes in the transfer function, suggesting a possible relationship between the two parameters. We suggest that the improved input-output relationship of neurons during optimal stimulation represents a novel mechanism that may contribute to the final sharp orientation selectivity of spike responses in the cortical cells. PMID- 11927690 TI - Laminar processing of stimulus orientation in cat visual cortex. AB - One of the most salient features to emerge in visual cortex is sensitivity to stimulus orientation. Here we asked if orientation selectivity, once established, is altered by successive stages of cortical processing. We measured patterns of orientation selectivity at all depths of the cat's visual cortex by making whole cell recordings with dye-filled electrodes. Our results show that the synaptic representation of orientation indeed changes with position in the microcircuit, as information passes from layer 4 to layer 2+3 to layer 5. At the earliest cortical stage, for simple cells in layer 4, orientation tuning curves for excitation (depolarization) and inhibition (hyperpolarization) had similar peaks (within 0-7 deg, n = 11) and bandwidths. Further, the sharpness of orientation selectivity covaried with receptive field geometry (r = 0.74) - the more elongated the strongest subregion, the shaper the tuning. Tuning curves for complex cells in layer 2+3 also had similar peaks (within 0-4 deg, n = 7) and bandwidths. By contrast, at a later station, layer 5, the preferred orientation for excitation and inhibition diverged such that the peaks of the tuning curves could be as far as 90 deg apart (average separation, 54 deg; n = 6). Our results support the growing consensus that orientation selectivity is generated at the earliest cortical level and structured similarly for excitation and inhibition. Moreover, our novel finding that the relative tuning of excitation and inhibition changes with laminar position helps resolve prior controversy about orientation selectivity at later phases of processing and gives a mechanistic view of how the cortical circuitry recodes orientation. PMID- 11927692 TI - Effects of prevailing hypoxaemia, acidaemia or hypoglycaemia upon the cardiovascular, endocrine and metabolic responses to acute hypoxaemia in the ovine fetus. AB - Although it is established that the fetus can successfully withstand a single, acute hypoxaemic challenge during gestation, little is known about what effects prevailing adverse intrauterine conditions might have on the fetal response to acute hypoxaemia. The aims of this study were therefore: (1) to characterise the effects of prevailing and sustained hypoxaemia, acidaemia or hypoglycaemia on the fetal cardiovascular responses to an episode of acute hypoxaemia; and (2) to determine the effects of these adverse intrauterine conditions on mechanisms mediating these cardiovascular responses. Thirty-three Welsh Mountain sheep fetuses were chronically instrumented (1-2 % halothane) between 117 and 125 days of gestation (term is ca 145 days) with amniotic and vascular catheters and with a transit-time flow probe around a femoral artery. The animals were divided retrospectively into four groups based upon post-surgical, sustained, basal blood oxygen (chronically hypoxaemic; P(a,O2), 17.3 +/- 0.5 mmHg; n = 8), glucose (chronically hypoglycaemic; blood glucose, 0.49 +/- 0.03 mmol l(-1); n = 6) and acid-base (chronically acidaemic; pH(a), 7.25 +/- 0.01; n = 5) status. Values for compromised fetuses were -2 S.D. from a group of control (n = 14) fetuses. At 130 +/- 4 days, a 1 h episode of acute, isocapnic hypoxaemia (9 % O(2) in N(2), to reduce carotid P(a,O2) to 12 +/- 1 mmHg) was induced in all fetuses by reducing the maternal inspired O(2) fraction (F(I,O2)). Fetal cardiovascular variables were recorded at 1 s intervals throughout the experimental protocol and arterial blood samples taken at appropriate intervals for biophysical (blood gases, glucose, lactate) and endocrine (catecholamines, vasopressin, cortisol, ACTH) measures. During acute hypoxaemia all fetuses elicited hypertension, bradycardia and femoral vasoconstriction. However, prevailing fetal compromise altered the cardiovascular and endocrine responses to a further episode of acute hypoxaemia, including: (1) enhanced pressor and femoral vasoconstriction; (2) greater increments in plasma noradrenaline and vasopressin during hypoxaemia; and (3) basal upward resetting of hypothalamic-pituitary-adrenal axis function. Only chronically hypoxaemic fetuses had significantly elevated basal concentrations of noradrenaline and enhanced chemoreflex function during acute hypoxaemia. These data show that prevailing adverse intrauterine conditions alter the capacity of the fetus to respond to a subsequent episode of acute hypoxaemia; however, the partial contributions of hypoxaemia, acidaemia or hypoglycaemia to mediating these responses can vary. PMID- 11927693 TI - Long lasting effects of rTMS and associated peripheral sensory input on MEPs, SEPs and transcortical reflex excitability in humans. AB - We tested the effect of repetitive transcranial magnetic stimulation (rTMS) over the motor cortex on the size of transcortical stretch and mixed nerve reflexes. Fourteen healthy subjects were investigated using either 25 min of 1 Hz rTMS or 30 min of 0.1 Hz rTMS paired with electrical stimulation of the motor point of the first dorsal interosseous muscle (FDI). Following treatment, we measured the effect on the size of: (1) EMG responses evoked in FDI by transcranial magnetic stimulation (MEPs), (2) somatosensory evoked potentials (SEPs) evoked by ulnar nerve stimulation, and (3) transcortical stretch or electrically elicited reflexes. rTMS at 1 Hz reduced the amplitude of both MEPs and long latency reflexes by 20-30 % for about 10 min after the end of stimulation. Short latency reflexes were unaffected. SEPs were not studied, as it has been shown previously that they are also suppressed. rTMS at 0.1 Hz paired with motor point stimulation (interstimulus interval of 25 ms) increased the amplitude of the MEP and the cortical components of the SEP (N20/P25 and later peaks) for up to 10 min. Long latency reflexes were facilitated with the same time course. We conclude that rTMS over the motor cortex either alone or in conjunction with peripheral inputs can decrease or increase the excitability of the sensory and motor cortex for short periods after the end of stimulation. These changes affect not only MEPs and SEPs but also EMG responses to more 'natural' inputs involved in transcortical stretch reflexes. PMID- 11927691 TI - Synaptic physiology of the flow of information in the cat's visual cortex in vivo. AB - Each stage of the striate cortical circuit extracts novel information about the visual environment. We asked if this analytic process reflected laminar variations in synaptic physiology by making whole-cell recording with dye-filled electrodes from the cat's visual cortex and thalamus; the stimuli were flashed spots. Thalamic afferents terminate in layer 4, which contains two types of cell, simple and complex, distinguished by the spatial structure of the receptive field. Previously, we had found that the postsynaptic and spike responses of simple cells reliably followed the time course of flash-evoked thalamic activity. Here we report that complex cells in layer 4 (or cells intermediate between simple and complex) similarly reprised thalamic activity (response/trial, 99 +/- 1.9 %; response duration 159 +/- 57 ms; latency 25 +/- 4 ms; average +/- standard deviation; n = 7). Thus, all cells in layer 4 share a common synaptic physiology that allows secure integration of thalamic input. By contrast, at the second cortical stage (layer 2+3), where layer 4 directs its output, postsynaptic responses did not track simple patterns of antecedent activity. Typical responses to the static stimulus were intermittent and brief (response/trial, 31 +/- 40 %; response duration 72 +/- 60 ms, latency 39 +/- 7 ms; n = 11). Only richer stimuli like those including motion evoked reliable responses. All told, the second level of cortical processing differs markedly from the first. At that later stage, ascending information seems strongly gated by connections between cortical neurons. Inputs must be combined in newly specified patterns to influence intracortical stages of processing. PMID- 11927696 TI - Epidemiology of pediatric injury-related primary care office visits in the United States. AB - OBJECTIVES: Unintentional injuries are a major cause of childhood mortality and morbidity in the United States. However, there is little epidemiologic information about pediatric injuries seen in primary care settings. The objective of this study was to characterize types and external causes of childhood injuries seen by primary care physicians and to compare the demographic and visit characteristics of children with injury-related visits (IRVs) and non-IRVs. METHODS: A stratified random sample survey of office-based practicing physicians in the National Ambulatory Medical Care Survey (1997 and 1998) was conducted. Injury-related primary care office visits were measured for patients who were <19 years (number of patient visits = 6358). RESULTS: Visits for childhood injuries composed >10 million primary care office visits per year (a rate of 13.8 visits per 100 person-years). The most common diagnoses assigned to these injuries were open wounds, sprains and strains, contusions, and superficial injuries such as abrasions and splinters. Leading external causes of these injuries were sports and overexertion, accidental falls, natural factors such as bites and stings, and cutting instruments. The single most common cause of pediatric injuries was sports/overexertion. Children who had IRVs were more likely to be older (odds ratio [OR]: 1.10/year of age; 95% confidence interval [CI]: 1.08-1.12), to be male (OR: 1.5; 95% CI: 1.2-1.9), and to reside in the West (OR: 1.9; 95% CI: 1.4 2.6) or in a rural area (OR: 1.4; 95% CI: 1.1-1.9). They were less likely to be Asian (OR: 0.2; 95% CI: 0.1-0.5) or Hispanic (OR: 0.5; 95% CI: 0.3-0.8). At the visit, children with IRVs were more likely to see a physician who was not their primary care physician (OR: 1.8; 95% CI: 1.4-2.3) and to see a family physician rather than a pediatrician (OR: 2.3; 95% CI: 1.8-2.9). CONCLUSIONS: IRVs account for a significant portion of primary care for children in the United States. Identifying potentially preventable external causes of injury and characteristics of children who are more likely to be injured is important both for injury prevention programs and for the education of physicians who care for children. PMID- 11927695 TI - Intramuscular fatty acid metabolism in contracting and non-contracting human skeletal muscle. AB - The present study was undertaken to investigate the fate of blood-borne non esterified fatty acids (NEFA) entering contracting and non-contracting knee extensor muscles of healthy young individuals. [U-(13)C]-palmitate was infused into a forearm vein during 5 h of one-legged knee extensor exercise at 40 % of maximal work capacity and the NEFA kinetics, oxidation and rate of incorporation into intramuscular triacylglycerol (mTAG) were determined for the exercising and the non-exercising legs. During 4 h of one-legged knee extensor exercise, mTAG content decreased by 30 % (P < 0.05) in the contracting muscle, whereas it was unchanged in the non-contracting muscle. The uptake of plasma NEFA, as well as the proportion directed towards oxidation, was higher in the exercising compared to the non-exercising leg, whereas the rate of palmitate incorporation into mTAG was fourfold lower (0.70 +/- 0.14 vs. 0.17 +/- 0.04 micromol (g dry wt)(-1) h( 1); P < 0.05), resulting in fractional synthesis rates of 1.0 +/- 0.2 and 3.8 +/- 0.9 % h(-1) (P < 0.01) for the contracting and non-contracting muscle, respectively. These findings demonstrate that mTAG in human skeletal muscle is continuously synthesised and degraded and that the metabolic fate of plasma NEFA entering the muscle is influenced by muscle contraction, so that a higher proportion is directed towards oxidation at the expense of storage in mTAG. PMID- 11927694 TI - Nitric oxide-dependent modulation of sympathetic neural control of oxygenation in exercising human skeletal muscle. AB - Nitric oxide (NO) attenuates alpha-adrenergic vasoconstriction in contracting rodent skeletal muscle, but it is unclear if NO plays a similar role in human muscle. We therefore hypothesized that in humans, NO produced in exercising skeletal muscle blunts the vasoconstrictor response to sympathetic activation. We assessed vasoconstrictor responses in the microcirculation of human forearm muscle using near-infrared spectroscopy to measure decreases in muscle oxygenation during reflex sympathetic activation evoked by lower body negative pressure (LBNP). Experiments were performed before and after NO synthase inhibition produced by systemic infusion of N(G)-nitro-L-arginine methyl ester (L NAME). Before L-NAME, LBNP at -20 mmHg decreased muscle oxygenation by 20 +/- 2 % in resting forearm and by 2 +/- 3 % in exercising forearm (n = 20), demonstrating metabolic modulation of sympathetic vasoconstriction. As expected, L-NAME increased mean arterial pressure by 17 +/- 3 mmHg, leading to baroreflex-mediated suppression of baseline muscle sympathetic nerve activity (SNA). The increment in muscle SNA in response to LBNP at -20 mmHg also was attenuated after L-NAME (before, +14 +/- 2; after, +8 +/- 1 bursts min(-1); n = 6), but this effect of L NAME was counteracted by increasing LBNP to -40 mmHg (+19 +/- 2 bursts min(-1)). After L-NAME, LBNP at -20 mmHg decreased muscle oxygenation similarly in resting (-11 +/- 3 %) and exercising (-10 +/- 2 %) forearm (n = 12). Likewise, LBNP at 40 mmHg decreased muscle oxygenation both in resting (-19 +/- 4 %) and exercising (-21 +/- 5 %) forearm (n = 8). These data advance the hypothesis that NO plays an important role in modulating sympathetic vasoconstriction in the microcirculation of exercising muscle, because such modulation is abrogated by NO synthase inhibition with L-NAME. PMID- 11927697 TI - Comparison of nasogastric and intravenous methods of rehydration in pediatric patients with acute dehydration. AB - OBJECTIVE: To assess the safety, efficacy, and cost-effectiveness of rapid nasogastric hydration (RNG) and rapid intravenous hydration (RIV) administered in the emergency department (ED) to young children suffering with uncomplicated, acute moderate dehydration. METHODS: Ninety-six children aged 3 to 36 months, who presented with signs and symptoms of uncomplicated, acute moderate dehydration caused by vomiting and/or diarrhea, presumed to be caused by viral gastroenteritis, were randomly assigned to receive either RNG with a standard oral rehydration solution or RIV with normal saline. Each solution was administered at a rate of 50 mL/kg of body weight, delivered over a 3-hour period in our urban pediatric ED. All participants were weighed pretreatment and posttreatment and underwent initial and final measurements of their serum electrolytes, blood urea nitrogen, creatinine, and glucose levels, along with urine chemistry and urine specific gravity. Telephone follow-up by completion of a standardized questionnaire was obtained approximately 24 hours after discharge from the ED. RESULTS: Ninety-two of 96 enrolled patients completed the study. Three patients failed treatment (2 RIV and 1 RNG) and were excluded and hospitalized because of severe, intractable vomiting, and 1 patient was withdrawn secondary to an intussusception. Among 92 evaluable patients, 2 were found to be severely dehydrated (>10% change in body weight) and were excluded from analysis, leaving 90 patients (RNG: N = 46 and RIV: N = 44), who completed the study. Both RNG and RIV were found to be a safe and efficacious means of treating uncomplicated, acute moderate dehydration in the ED. Determinations of electrolytes, blood urea nitrogen, creatinine, or glucose were not found to be of value on an intent-to-treat basis in the care of these patients. The urine specific gravity and incidence of ketonuria declined from levels commensurate with moderate dehydration in the RNG group, but not as consistently so in the RIV group. Both RNG and RIV were substantially less expensive to administer than standard care with intravenous fluid deficit therapy in-hospital, and RNG was more cost-effective to administer over RIV in the outpatient setting. CONCLUSION: RNG and RIV administered in the ED are safe, efficacious, and cost-effective alternatives to the standard treatment for uncomplicated, acute moderate dehydration in young children. RNG is as efficacious as RIV, is no more labor intensive than RIV, and is associated with fewer complications. In addition, we found that most routine laboratory testing is of little value in these patients and should be avoided, except when clearly clinically indicated. PMID- 11927698 TI - Behavior and development of preschool children born to adolescent mothers: risk and 3-generation households. AB - OBJECTIVE: To investigate whether living in a 3-generation household (grandmother mother-child) is associated with fewer behavior problems and better cognitive development among preschool children of mothers who gave birth during adolescence and whether it protects children from the behavior and developmental problems associated with maltreatment and maternal depression. DESIGN: Cohort study. SETTING: Participants included low-income families recruited from 4 sites: East, Northwest, Midwest, and South, who are part of LONGSCAN, a longitudinal study of children's health, development, and maltreatment. PARTICIPANTS: One hundred ninety-four mothers who were adolescents (less than age 19) at delivery. Data were gathered when children were 4 to 5 years of age. Twenty-six percent of the children lived in 3-generation households, 39% had a history of maltreatment, and 32% of the mothers had depression scores in the clinical range. MAIN OUTCOME MEASURES: Child behavioral problems were measured with the Child Behavior Checklist, completed by the mother, and child developmental status was assessed with the Battelle Developmental Inventory Screening Test, administered by research assistants. RESULTS: Multiple regression analyses revealed that children who had been reported for maltreatment and had mothers with depressive symptoms had more externalizing behavior problems, compared with children who experienced neither risk or only 1 risk. However, when residential status was considered, children with the greatest number of externalizing behavior problems were those who experienced both maltreatment and maternal depressive symptoms and lived in 3 generation households. Children who had been reported for maltreatment or had mothers with depressive symptoms were more likely to have internalizing problems, compared with children with neither risk. Residential status was not related to children's internalizing behavior problems or cognitive development. CONCLUSIONS: Living in a 3-generation household did not protect preschool children from the behavior problems associated with maltreatment and depression. In contrast, living in a 3-generation household was associated with more behavior problems among the highest risk group of children--those who had been maltreated and had mothers with symptoms of depression. Although 3-generation families may provide an important source of support and stability for adolescent mothers and their infants early in the parenting process, it may not be advisable to rely on 3 generation households as young mothers enter adulthood, particularly among those with a history of maltreatment or depression. Children with the fewest number of behavior problems were living with their mothers in their own household (often with the father), had not been maltreated, and had mothers with few symptoms of depression. PMID- 11927699 TI - Self-esteem as a predictor of initiation of coitus in early adolescents. AB - OBJECTIVE: To examine the role of self-esteem in predicting coitus initiation in a population of early adolescents. METHODS: Questionnaires assessing coital status, self-esteem, and pubertal maturation were collected from 188 participants who had not engaged in coitus by enrollment. Data were collected longitudinally while the participants were in the seventh and ninth grade. Ages ranged from 12 to 14 (mean: 12.50; standard deviation: 0.57) at Time 1 and from 14 to 16 (mean: 14.30; standard deviation: 0.49) at Time 2. RESULTS: Boys with higher self-esteem ratings at Time 1 were more likely to initiate intercourse by Time 2. Girls with higher self-esteem at Time 1 were more likely to remain virgins than girls with lower self-esteem. Pubertal status was unrelated to initiation of coitus in this sample. CONCLUSIONS: Self-esteem, regardless of pubertal status, predicted coitus transition differentially in boys and girls. Results from this longitudinal study seem to fit within traditional problem behavior theory. PMID- 11927700 TI - Infant acceptance of breast milk after maternal exercise. AB - OBJECTIVE: Previous research reported that breast milk lactic acid (LA) levels increase after lactating women complete a bout of exhaustive exercise, resulting in poor infant acceptance of the postexercise breast milk. This highly publicized finding may not apply to more practical, everyday exercise conditions of lactating women. The purpose of the present study was to reexamine the composition and infant acceptance of postexercise breast milk while controlling maternal diet, exercise intensity, and the method, timing, and assessment of infant feeding. RESEARCH DESIGN AND METHODS: Twenty-four women, 2 to 4 months' postpartum, completed 3 test sessions: a maximal oxygen uptake test, a 30-minute bout of moderate exercise, and a resting control session. One hour before and 1 hour after each session, participants fully expressed their milk, placed it in a bottle familiar to the infant, fed their infant, and rated their infant's acceptance of the milk. Each feeding was videotaped and viewed individually by 3 lactation consultants who rated infant acceptance; consultants were blinded to the test sessions. Milk was analyzed for LA and infant milk consumption was measured. RESULTS: There were no differences in presession versus postsession values for maternal skin temperature, breast milk temperature, and infant milk acceptance as judged by either the mothers or lactation consultants. These results prevailed despite a small but significant increase in breast milk LA premaximal versus postmaximal exercise (0.09 vs 0.21 mM, respectively); there was no difference in milk LA premoderate versus postmoderate exercise, or prerest versus postrest. CONCLUSION: These data support the hypothesis that moderate or even high-intensity exercise during lactation does not impede infant acceptance of breast milk consumed 1 hour postexercise. PMID- 11927701 TI - Breastfeeding is analgesic in healthy newborns. AB - CONTEXT: This study identifies a behavioral and nonpharmacologic means of preventing newborn pain. OBJECTIVE: To determine whether breastfeeding is analgesic in newborn infants undergoing heel lance-a routine, painful, hospital procedure. DESIGN: A prospective, randomized, controlled trial. SETTING: Hospital maternity services at Boston Medical Center, Boston, Massachusetts, and Beverly Hospital, Beverly, Massachusetts. PARTICIPANTS: A random sample of 30 full-term, breastfed infants. INTERVENTIONS: Infants in the intervention group were held and breastfed by their mothers during heel lance and blood collection procedures for the Newborn Screening Program Blood Test. Infants in the control group experienced the same blood test while receiving the standard hospital care of being swaddled in their bassinets. OUTCOMES MEASURES: Crying, grimacing, and heart rate differences were analyzed between the breastfeeding and the control infants before, during, and after blood collection. RESULTS: Crying and grimacing were reduced by 91% and 84%, respectively, from control infant levels during the blood collection. Heart rate was also substantially reduced by breastfeeding. CONCLUSIONS: Breastfeeding is a potent analgesic intervention in newborns during a standard blood collection. PMID- 11927702 TI - Use of sleep aids during the first year of life. AB - OBJECTIVE: In an attempt to foster self-soothing during the night, a novel sleep aid infused with maternal odor was introduced to 4 groups of infants ranging in age from 3 to 12 months. Infants' use of parent-provided sleep aids also was examined. METHODOLOGY: Nighttime sleep and waking behaviors were videotaped for 2 consecutive nights on 3 occasions over a 3-month interval. Using all-night video recording, the study examined the infant's use of a novel sleep aid and parent provided sleep aids during sleep onset and after nighttime awakenings. RESULTS: Results indicated that infants of different ages differed in the types of sleep aids used when falling asleep either at the beginning of the night or after awakenings in the middle of the night. More 3-month-olds used their thumbs/fingers/hands, whereas more 6-month-olds used soft objects. The 6-month olds were most likely to use the novel sleep aid. Almost all of the infants at all 4 ages used some type of object during the night. Intra-individual analyses showed that infants tended to change their pattern of sleep aid use over the 3 month study period. CONCLUSIONS: The data provide evidence that infants during the first year of life use sleep aids frequently and interchangeably rather than a specific favorite object. PMID- 11927703 TI - Terbinafine in the treatment of Trichophyton tinea capitis: a randomized, double blind, parallel-group, duration-finding study. AB - OBJECTIVES: Terbinafine has been shown to be effective in tinea capitis, using different treatment durations. However, no direct comparison of treatment duration has previously been investigated. This randomized, double-blind, parallel-group, multicenter study was designed to assess the effect of terbinafine treatment duration on the outcome of Trichophyton tinea capitis in a North American population. METHODS: A total of 176 patients with a clinical diagnosis of tinea capitis were enrolled in this study and treated with oral terbinafine (3-6 mg/kg/d) for 1, 2, or 4 weeks. All patients were to be followed until week 12. A total of 159 patients had culture-confirmed tinea capitis attributable to Trichophyton species and constituted the intent-to-treat population used for efficacy analysis (50, 55, and 54 patients in the 1-, 2-, and 4-week arms, respectively). RESULTS: At the end of study, effective treatment, defined as negative culture and low scores on signs and symptoms, was achieved in 56%, 69%, and 65% of patients who were treated with terbinafine for 1, 2, and 4 weeks, respectively. A negative culture was achieved in 60%, 76%, and 72%, respectively. Overall, the efficacy data showed that both the 2- and 4-week treatment regimens are clinically superior to the 1-week regimen. Terbinafine was well tolerated, and the incidence of adverse events showed no relationship to the duration of therapy. CONCLUSION: When efficacy, cost, and compliance are taken into consideration, 2 weeks of terbinafine therapy appears to be the optimal treatment duration for patients with Trichophyton tonsurans tinea capitis. PMID- 11927704 TI - Infant sleep placement after the back to sleep campaign. AB - OBJECTIVES: The Back to Sleep campaign has been credited with recent declines in the incidence of sudden infant death syndrome. Using survey data for the 1996 1998 birth cohorts, this epidemiologic study examines infant sleep position in a large, population-based sample. DATA AND METHODS: Data concerning infant sleep position are drawn from the 1996-1998 Pregnancy Risk Assessment Monitoring System for 15 states. Weighted multiple logistic regression analysis is used to examine correlates of infant sleep position. RESULTS: The prevalence of prone infant sleeping significantly declined between 1996 and 1998 (adjusted odds ratio [AOR] = 0.70; 95% confidence interval [CI] = [0.63, 0.78]). African Americans were more likely than non-Hispanic whites to sleep prone, (AOR = 1.45; 95% CI = 1.33,1.59), and were less likely to sleep supine (AOR = 0.52; 95% CI = 0.48, 0.57). Hispanic/Latinos were less likely overall than non-Hispanic whites to sleep prone (AOR = 0.81; 95% CI = 0.69, 0.95), but were also less likely to sleep supine (AOR = 0.78; 95% CI = 0.69, 0.87). Adherence to sleep position recommended by the American Academy of Pediatrics increased sharply among Hispanic/Latino infants. Very low birth weight infants and infants in larger families were less likely to sleep in the recommended supine position. Infants born between 1001 and 1500 g (AOR = 0.67; 95% CI = 0.57, 0.79), and extremely low birth weight infants between 500 and 1000 g (AOR = 0.57; 95% CI = 0.45, 0.72) were especially unlikely to sleep supine. Infants in households with more than 3 other children (AOR = 1.72; 95% CI = 1.08, 2.74) were more likely to sleep prone. CONCLUSIONS: The prevalence of supine infant sleep increased between 1996 and 1998. Low adherence to sleep position recommendations of the American Academy of Pediatrics among African Americans, very low birth weight infants, and infants in large families remain public health concerns. PMID- 11927705 TI - Household composition and risk of fatal child maltreatment. AB - OBJECTIVE: Approximately 2000 children die annually in the United States from maltreatment. Although maternal and child risk factors for child abuse have been identified, the role of household composition has not been well-established. Our objective was to evaluate household composition as a risk factor for fatal child maltreatment. METHODOLOGY: Population-based, case-control study using data from the Missouri Child Fatality Review Panel system, 1992-1994. Households were categorized based on adult residents' relationship to the deceased child. Cases were all maltreatment injury deaths among children <5 years old. Controls were randomly selected from natural-cause deaths during the same period and frequency matched to cases on age. The main outcome measure was maltreatment death. RESULTS: Children residing in households with adults unrelated to them were 8 times more likely to die of maltreatment than children in households with 2 biological parents (adjusted odds ratio [aOR]: 8.8; 95% confidence interval [CI]: 3.6-21.5). Risk of maltreatment death also was elevated for children residing with step, foster, or adoptive parents (aOR: 4.7; 95% CI: 1.6-12.0), and in households with other adult relatives present (aOR: 2.2; 95% CI: 1.1-4.5). Risk of maltreatment death was not increased for children living with only 1 biological parent (aOR: 1.1; 95% CI: 0.8-2.0). CONCLUSIONS: Children living in households with 1 or more male adults that are not related to them are at increased risk for maltreatment injury death. This risk is not elevated for children living with a single parent, as long as no other adults live in the home. PMID- 11927706 TI - Role of L-carnitine in apnea of prematurity: a randomized, controlled trial. AB - OBJECTIVE: Carnitine is thought to be a conditionally essential biological cofactor for premature infants. A preliminary study suggested that carnitine could significantly reduce apnea of prematurity. The objective of this study was to evaluate critically the role of carnitine in idiopathic apnea of prematurity and to determine whether the use of carnitine would facilitate discontinuation of mechanical ventilatory support, shorten the duration of ventilatory support, and reduce the amount of time that such infants are exposed to both mechanical ventilation and oxygen. We also wanted to determine the effects of supplemental carnitine on weight gain, time to regain birth weight, time to achieve full enteral feedings, and length of hospital stay. METHODS: A prospective, randomized, blinded trial was conducted on 44 preterm infants who were from the same neonatal intensive care unit and who were < or =32 weeks' gestational age with a postnatal age <48 hours and a birth weight <1500 g and required total parenteral nutrition (TPN). Infants were randomized to receive carnitine supplementation or placebo without crossover. Carnitine-supplemented infants received 30 mg/kg/d carnitine in their TPN until the they were tolerating 120 mL/kg/d enteral feedings, and then they received 30 mg/kg/d oral carnitine. The placebo group received TPN without supplemental carnitine; when they tolerated 120 mL/kg/d enteral feedings, they received an oral placebo. The 2 groups continued on their respective supplemental carnitine or placebo until 34 weeks' adjusted age, at which time the study period was completed. Twelve-hour cardiorespiratorygrams to record heart rate, respiratory impedance, and oxygen saturation, and a nasal thermistor to detect expiratory airflow were performed every 4 days on 3 occasions and at 30 and 34 weeks' adjusted age. Plasma carnitine levels were measured at day 14. RESULTS: There were no significant differences between the 2 groups in the occurrence of apnea as detected by cardiorespiratorygram or nursing observation. There were no significant differences between the groups in regard to total days on ventilator, days of nasal continuous positive airway pressure, time to regain birth weight, time to reach enteral feedings of 120 mL/kg/d, discharge weight, adjusted age at discharge, need for oxygen at 28 days' and 36 weeks' adjusted age, or length of stay. The plasma carnitine level was a median of 15.5 micromol/L (range: 7.6 30.5) for the placebo infants compared with a median of 195.3 micromol/L (range: 71.7-343.6) for the carnitine infants. CONCLUSIONS: In this blinded, randomized, placebo-controlled study, we found that infants who received supplemental carnitine did not demonstrate any reduction in apnea of prematurity, ventilator or nasal continuous positive airway pressure days, or the need for supplemental oxygen therapy. Although carnitine may be of significant nutritional benefit for very low birth weight infants, our study does not support its use to reduce apnea of prematurity or decrease dependence on mechanical ventilation. PMID- 11927707 TI - Disparities in infant health among American Indians and Alaska natives in US metropolitan areas. AB - OBJECTIVE: To determine geographic variation in urban American Indian and Alaska Native (AI/AN) rates of infant mortality, low birth weight, prenatal care use, and maternal-child health care service availability. METHODS: This was a retrospective cohort study using data from the 1989 to 1991 birth-death linked database from the National Center for Health Statistics. We examined births from metropolitan areas with a minimum of 300 AI/AN births during the study period. Key outcomes of interest included rates of low birth weight, neonatal mortality, postneonatal mortality, and women receiving inadequate prenatal care using the modified Kessner index. To determine the type of health services tailored to AI/AN mothers residing in these urban areas, we conducted a telephone survey of the 36 urban Indian health programs operating in 1997 using a semistructured survey. Items in the survey included questions about the availability of prenatal and infant health care. RESULTS: During the 1989 to 1991 study period, there were 72 730 singleton births to AI/AN mothers and/or fathers residing in urban areas, representing 49% of all AI/AN births in the United States. Overall 14.4% of urban AI/AN births were to women who received inadequate care during pregnancy, 5.7% of pregnancies resulted in low birth weight infants, and 11.0 infants died per 1000 live births. Death rates for the neonatal period (5.5 per 1000 births) and postneonatal period (5.4 per 1000 births) were similar. Marked disparity in these indicators exist between pregnancies to AI/AN and white women. Among the 54 metropolitan areas, 46 had a rate ratio (AI/AN: white) for inadequate care of > or =1.5 (range: 0.9-8.5). The mean rate ratios for neonatal and postneonatal mortality were 1.6 (range: 0.3-4.0) and 2.0 (range: 0.5-5.5). There was also considerable geographic variation of AI/AN mortality rates between metropolitan areas in all of the outcomes studied. All of the 20 metropolitan areas with the highest birth counts had some type of direct medical care or outreach services available from an urban clinic targeted toward AI/AN patients. CONCLUSIONS: Considerable variation also exists among rates of AI/ANs between metropolitan areas. Disparity exists in rates of perinatal outcomes between AI/ANs and whites living in the same metropolitan areas Although AI/AN urban health programs exist in most cities with large birth counts, it seems that many have inadequate resources to meet existing needs to improve perinatal outcomes and infant health. PMID- 11927708 TI - Prevalence, correlates, and trajectory of television viewing among infants and toddlers. AB - OBJECTIVES: Recognizing the negative effects of television on children, the American Academy of Pediatrics (AAP) recommends that children 2 years and older watch <2 hours of television per day and that children younger than 2 years watch no television. However, relatively little is known about the amount of television viewed by infants and toddlers. The objective of this study was to describe the prevalence and correlates of television viewing that exceeds the AAP guidelines for 0- to 35-month-olds and to examine the trajectory of a child's viewing over time. METHODS: Data from the National Longitudinal Survey of Youth, 1990 to 1998, were used to analyze reported television viewing at 0 to 35 months of age and to follow the trajectory of a child's viewing from infancy through age 6. Logistic regression models were used to determine risk factors associated with greater television viewing at 0 to 35 months and the association of early viewing habits with school-age viewing. RESULTS: Seventeen percent of 0- to 11-month-olds, 48% of 12- to 23-month-olds, and 41% of 24- to 35-month-olds were reported to watch more television than the AAP recommends. Compared with college graduates, less educated women were more likely to report that their children watched more television than recommended. Children who watched >2 hours per day at age 2 were more likely to watch >2 hours per day at age 6 (odds ratio: 2.7; 95% confidence interval: 1.8-3.9), controlling for maternal education, race, marital status and employment, household income, and birth order. CONCLUSIONS: A substantial number of children begin watching television at an earlier age and in greater amounts than the AAP recommends. Furthermore, these early viewing patterns persist into childhood. Preventive intervention research on television viewing should consider targeting infants and toddlers and their families. PMID- 11927709 TI - Significant selective head cooling can be maintained long-term after global hypoxia ischemia in newborn piglets. AB - OBJECTIVE: Selective head cooling (SHC) combined with mild body cooling is currently being evaluated as a potentially therapeutic option in the management of neonatal hypoxic-ischemic encephalopathy. It is proposed that SHC enables local hypothermic neuroprotection while minimizing the deleterious side effects of systemic hypothermia. However, there is little evidence that it is possible to cool the brain more than the body for a prolonged period of time. The aim of this study was to examine whether the brain (T(deep brain)) could be cooled to below the rectal temperature (T(rectal)) in our piglet hypoxia ischemia (HI) model for a period of 24 hours, using a head-cooling cap. METHODS: Eight anesthetized piglets (median age: 15 hours) had subdural and intracerebral basal ganglia temperature probes inserted. After a 45-minute global HI insult (known to produce permanent brain damage), SHC using a cap perfused with cold water (5 degrees C-24 degrees C) combined with overhead body heating to maintain T(rectal) at 34 to 35 degrees C was performed for 24 hours. RESULTS: The piglets were cooled to a median T(rectal) of 35.0 degrees C (interquartile range [IQR]: 34.7-35.3) for 24 hours. During this time, the median T(deep brain) was 31.4 degrees C (IQR: 30 degrees C-32.2 degrees C), with a median T(rectal) to T(deep brain) gradient of 3.4 degrees C (IQR: 2.7 degrees C-4.8 degrees C). At the end of the cooling period, this gradient was still maintained at a median of 3.3 degrees C (IQR: 2.9 degrees C-3.7 degrees C). The ability to obtain the gradient was not influenced by the size of the piglet (1300-1840 g). Cap cooling lowered scalp temperature (T(scalp)) to a median of 24.9 degrees C (IQR: 22.2 degrees C-29.2 degrees C) and subdural temperature to a median of 28.1 degrees C (IQR: 25.8 degrees C-29.5 degrees C) but did not result in either skin injury or superficial brain hemorrhage. There was no clinically useful correlation between T(scalp) and T(deep brain) or between T(scalp) and T(subdural). CONCLUSIONS: This study using our piglet HI model shows that it is possible by means of a head-cooling cap to cool the brain more than the body for a 24-hour period while keeping the core temperature mildly hypothermic. However, we were unable to predict temperatures inside the brain using surface temperature probes on the head. PMID- 11927710 TI - Placental features in preterm infants with periventricular leukomalacia. AB - OBJECTIVE: Evaluation of the placenta provides some important insights into pathophysiologic changes that take place during the prenatal and intrapartum process. We investigated the relationship between placental findings and periventricular leukomalacia (PVL) to obtain a better understanding of its cause. METHODS: Thirty-two preterm infants with PVL delivered before 34 weeks' gestation, between 1990 and 1999, were classified into 4 groups according to the onset of brain injury assumed from ultrasonographic presentation and clinical course: 2 Antenatal, 22 Peripartum, 5 Postnatal, and 3 in an unknown time of onset group. We evaluated the gross and histopathologic features of the placentas of each group and compared them with those of a control group matched by birth weight and gestational age in terms of the frequency of major placental findings. Potential confounding factors were controlled in logistic regression analyses. RESULTS: Gross lesions with disturbance of uteroplacental circulation, including massive retroplacental hematoma, extensive infarction or thrombosis, and marked basal or perivillous fibrin deposition, were observed more frequently in the Antenatal + Peripartum combined subgroup than in the controls (41.7% vs 13.7%). Placentas from the Antenatal + Peripartum subgroup also demonstrated a significantly higher frequency of ischemic changes in villi, based on histopathologic examination, as compared with the control group (54.2% vs 13.7%). These associations remained after adjustment for confounding factors in logistic regression analyses (odds ratio: 4.04, 95% confidence interval: 1.40-11.67; and odds ratio: 7.28, 95% confidence interval: 2.50-21.20; respectively). Frequencies of chorioamnionitis and twin placentation tended to be higher in PVL cases than in the controls, although the differences were not statistically significant (46.9% vs 37.9%, 37.5% vs 20.0%, respectively). CONCLUSIONS: These results suggest that disturbed placental circulation underlies the development of PVL in the majority of cases with prenatal and peripartum brain injury. In chorioamnionitis cases, certain additional factors were suggested in the genesis of PVL. Thus, placental examination is essential for elucidating the pathophysiologic changes leading to PVL in the perinatal process. PMID- 11927711 TI - Where do children with complex chronic conditions die? Patterns in Washington State, 1980-1998. AB - OBJECTIVE: Little is known about factors that influence whether children with chronic conditions die at home. We sought to test whether deaths attributable to underlying complex chronic conditions (CCCs) were increasingly occurring at home and to determine what features were associated with home deaths. DESIGN: A retrospective case series was conducted of all deaths that occurred to children age 0 to 18 years in Washington state from 1980 to 1998 using death certificate data, augmented with 1990 US Census data regarding median household income by zip code in 1989, to determine the site of death. RESULTS: Of the 31 455 deaths identified in infants, children, and adults younger than 25 years, 52% occurred in the hospital, 17.2% occurred at home, 8.5% occurred in the emergency department or during transportation, 0.4% occurred in nursing homes, and 21.7% occurred at other sites. Among children who died as a result of some form of CCC (excluding injury, sudden infant death syndrome, and non-CCC medical conditions), the percentage of cases younger than 1 year who died at home rose slightly from 7.8% in 1980 to 11.6% in 1998, whereas the percentage of older children and young adults who had a CCC and died at home rose substantially from 21% in 1980 to 43% in 1998. Children who had lived in more affluent neighborhoods were more likely to have died at home. Using leukemia-related deaths as a benchmark, deaths as a result of congenital, genetic, neuromuscular, and metabolic conditions and other forms of cancer all were more likely to have occurred at home. Significant variation in the likelihood of home death, not explained by the individual attributes of the cases, also existed across the 39 counties in Washington state. CONCLUSIONS: Children who die with underlying CCCs increasingly do so at home. Age at death, specific condition, local area affluence, and the location of home all influence the likelihood of home death. These findings warrant additional study, as they have implications for how we envision pediatric palliative care, hospice, and other supportive services for the future. PMID- 11927712 TI - Trends in postneonatal aspiration deaths and reclassification of sudden infant death syndrome: impact of the "Back to Sleep" program. AB - OBJECTIVE: The introduction of the "Back to Sleep" campaign for the prevention of sudden infant death syndrome (SIDS) brought with it concern that there might be an increase in the incidence of aspiration-related deaths. The objective of this analysis was to describe the trends in postneonatal mortality and proportionate mortality ratios for the United States for the years 1991 to 1996 for aspiration related deaths and other causes to which a SIDS death could conceivably be reclassified. METHODS: Linked birth and infant death vital statistic files for the United States were used for the years 1991, 1995, and 1996. US Vital Statistic Mortality files for the years 1992, 1993, and 1994 were used because of the absence of linked files for those years. RESULTS: The overall postneonatal mortality rate between 1991 and 1996 declined 21.9%, whereas the SIDS rate declined 38.9%. The proportion of the postneonatal mortality (PNPMR) contributed by SIDS declined from 37.1% in 1991 to 28.8% in 1996. There was no significant increase in the PNPMR for aspiration, asphyxia, or respiratory failure. There was, however, a significant increase in the PNPMR for suffocation in bed or cradle from 0.9 to 1.3. CONCLUSIONS: These data show no evidence of an increased risk of death from aspiration as a result of the "Back to Sleep" program. Although there has been an increase in the proportion of postneonatal mortality attributable to suffocation, this represents a very small proportion of postneonatal mortality and thus potentially a very small number of SIDS deaths reclassified as suffocation. PMID- 11927713 TI - Interchangeability of 2 diphtheria-tetanus-acellular pertussis vaccines in infancy. AB - OBJECTIVE: Currently, 4 diphtheria-tetanus-acellular pertussis (DTaP) vaccines are licensed for pediatric use in the United States, and 2 are commercially available. Although a single manufacturer's DTaP vaccine should be used for all 3 doses of the primary immunization series, some circumstances result in infants receiving DTaP vaccines from more than 1 manufacturer. The purpose of this study was to evaluate the safety and immunogenicity of a mixed sequence of 2 different DTaP vaccines. METHODS: In this multicenter, observer-blinded, controlled study, 449 infants were randomized into 1 of 3 groups (1:1:1 ratio) to receive Tripedia at 2, 4, and 6 months of age (control group); Tripedia at 2 and 4 months of age and Infanrix at 6 months of age; or Tripedia at 2 months and Infanrix at 4 and 6 months of age. Other vaccines were administered concurrently as separate injections according to the recommended childhood immunization schedule. Safety was monitored closely, and standard enzyme immunoassays were used to measure antibody concentrations to each antigen of the DTaP vaccines. RESULTS: The rates of injection-site and systemic adverse events were similar in each study group, and there were no clinically significant differences among groups after any dose. Infants in all 3 groups responded well to each antigen contained in both vaccines, with 97% to 100% seroprotection or vaccine response rates after the 3 dose primary series. Postvaccination geometric mean antibody concentrations and seroprotection or vaccine response rates to nearly all vaccine antigens were as high or higher in the mixed-sequence groups as in the control group. CONCLUSION: Initiating the primary immunization series with 1 or 2 doses of Tripedia and completing the 3-dose series with Infanrix is as safe and at least as immunogenic as administering Tripedia for all 3 doses. PMID- 11927714 TI - Detection of pulsus paradoxus associated with large pericardial effusions in pediatric patients by analysis of the pulse-oximetry waveform. AB - OBJECTIVE: To determine whether analysis of the pulse-oximetry waveform can be useful in detecting the pulsus paradoxus associated with large pericardial effusions in pediatric patients. METHODS: A retrospective review of charts of 8 pediatric patients (age range: 5-19 years) who had echocardiographic evidence of large pericardial effusion, subsequently underwent pericardiocentesis, and had pulse-oximetry waveform tracings obtained before and after pericardiocentesis within an 18-month period was conducted in 2 tertiary-care pediatric intensive care units. We analyzed the pulse-oximetry waveform tracings for the presence of a pulsus paradoxus. Other abstracted data included clinical evidence of tamponade, echocardiographic findings, and the volume of pericardial fluid aspirated. RESULTS: Before pericardiocentesis, a decrease in the highest value of the upper plethysmographic peak of the pulse-oximetry waveform was observed during inspiration in each patient. Echocardiographic evidence of large pericardial effusion with compromised cardiac filling was also present in each patient. Only 6 of these patients had clinical evidence of cardiac tamponade at that time, 4 with a documented pulsus paradoxus using standard methods of blood pressure analysis. After pericardiocentesis, the inspiratory fall in the highest value of the upper plethysmographic peak of the pulse-oximetry waveform lessened in every patient. Echocardiography documented a decrease in the size of the effusion and resolution of the compromised cardiac filling in every patient. CONCLUSIONS: Analysis of pulse-oximetry waveforms may be a widely available, easily interpretable, and reliable method of detecting the pulsus paradoxus associated with large pericardial effusions in pediatric patients. PMID- 11927715 TI - Lactobacillus therapy for acute infectious diarrhea in children: a meta-analysis. AB - OBJECTIVE: Childhood diarrhea accounts for substantial morbidity and mortality worldwide. Multiple studies in children have shown that Lactobacillus, administered orally, may have antidiarrheal properties. We conducted a meta analysis of randomized, controlled studies to assess whether treatment with Lactobacillus improves clinical outcomes in children with acute infectious diarrhea. METHODS: Studies were sought in bibliographic databases of traditional biomedical as well as complementary and alternative medicine literature published from 1966 to 2000. Search terms were "competitive inhibition," "diarrhea," "gastroenteritis," "Lactobacillus," "probiotic," "rotavirus," and "yog(h)urt." We included studies that were adequately randomized, blinded, controlled trials in which the treatment group received Lactobacillus and the control group received an adequate placebo and that reported clinical outcome measures of diarrhea intensity. These inclusion criteria were applied by blind review and consensus. The original search yielded 26 studies, 9 of which met the criteria. Multiple observers independently extracted study characteristics and clinical outcomes. Data sufficient to perform meta-analysis of the effect of Lactobacillus on diarrhea duration and diarrhea frequency on day 2 were contained in 7 and 3 of the included studies, respectively. RESULTS: Summary point estimates indicate a reduction in diarrhea duration of 0.7 days (95% confidence interval: 0.3-1.2 days) and a reduction in diarrhea frequency of 1.6 stools on day 2 of treatment (95% confidence interval: 0.7-2.6 fewer stools) in the participants who received Lactobacillus compared with those who received placebo. Details of treatment protocols varied among the studies. A preplanned subanalysis suggests a dose effect relationship. CONCLUSION: The results of this meta-analysis suggest that Lactobacillus is safe and effective as a treatment for children with acute infectious diarrhea. PMID- 11927716 TI - Diagnosis and management of suspected cases of bioterrorism: a pediatric perspective. AB - Since October 3, 2001, the Centers for Disease Control and Prevention and other organizations have been investigating potential bioterrorist-related anthrax cases. The pediatrician may be faced with complex issues related to diagnosis and treatment of illnesses caused by intentionally released biological agents. The agents that pose a major potential bioterrorist threat are reviewed by the clinical syndromes they produce: acute respiratory distress with fever, influenza like illnesses, acute rash with fever, neurologic syndromes, and blistering syndromes. Specific and detailed diagnostic, treatment, and prophylaxis information is provided for anthrax, plague, tularemia, smallpox, botulism, viral hemorrhagic fevers, and other diseases. In cases of suspected bioterrorism, the pediatrician must be able to obtain diagnostic and treatment information efficiently and expeditiously. The system controlling the interaction between public and nonpublic health laboratories in suspected cases of bioterrorism is described. Finally, information regarding emergency contacts and links to educational resources is provided. PMID- 11927718 TI - Clinical practice guideline: diagnosis and management of childhood obstructive sleep apnea syndrome. AB - This clinical practice guideline, intended for use by primary care clinicians, provides recommendations for the diagnosis and management of obstructive sleep apnea syndrome (OSAS). The Section on Pediatric Pulmonology of the American Academy of Pediatrics selected a subcommittee composed of pediatricians and other experts in the fields of pulmonology and otolaryngology as well as experts from epidemiology and pediatric practice to develop an evidence base of literature on this topic. The resulting evidence report was used to formulate recommendations for the diagnosis and management of childhood OSAS. The guideline contains the following recommendations for the diagnosis of OSAS: 1) all children should be screened for snoring; 2) complex high-risk patients should be referred to a specialist; 3) patients with cardiorespiratory failure cannot await elective evaluation; 4) diagnostic evaluation is useful in discriminating between primary snoring and OSAS, the gold standard being polysomnography; 5) adenotonsillectomy is the first line of treatment for most children, and continuous positive airway pressure is an option for those who are not candidates for surgery or do not respond to surgery; 6) high-risk patients should be monitored as inpatients postoperatively; 7) patients should be reevaluated postoperatively to determine whether additional treatment is required. This clinical practice guideline is not intended as a sole source of guidance in the evaluation of children with OSAS. Rather, it is designed to assist primary care clinicians by providing a framework for diagnostic decision-making. It is not intended to replace clinical judgment or to establish a protocol for all children with this condition and may not provide the only appropriate approach to this problem. PMID- 11927717 TI - Real versus theoretical: assessing the risks and benefits of postponing the hepatitis B vaccine birth dose. PMID- 11927719 TI - CHOICE Study Group Trial. PMID- 11927720 TI - Respiratory instability of term and near-term healthy newborn infants in car safety seats. PMID- 11927721 TI - Role of carbon monoxide and nitric oxide in newborn infants with postasphyxial hypoxic-ischemic encephalopathy. PMID- 11927722 TI - Meta-analysis of postnatal steroid use challenged. PMID- 11927723 TI - Definition and diagnostic criteria for acute otitis media. PMID- 11927724 TI - High-flow nasal cannula versus nasal continuous positive airway pressure in the management of apnea of prematurity. PMID- 11927725 TI - Poor readability of written asthma management plans found in national guidelines. AB - OBJECTIVE: Written asthma management plans (WAMPs) for patients constitute a key component of current national asthma guidelines, but it is not known whether these plans meet the readability standards (of fifth-grade level or lower) recommended by health education experts. The objective of this study was to assess whether WAMPs presented in national guidelines are written at or below a fifth-grade reading level. METHODS: We used readability software to analyze 10 WAMPs. These included 7 from the national guidelines, 1 from the World Health Organization, and 2 local ones. RESULTS: The grade levels for the WAMPs ranged from 4.9 to 9.2. None of the national plans achieved a grade level equal to or below the fifth grade. However, the other plans had grade levels of 4.9, 5.7, and 5.9. The mean grade level for the national plans was 8.1, whereas the mean for the other 3 plans was 5.5. The difference between these 2 sets of WAMPs was significant. CONCLUSION: WAMPs presented as part of the national guidelines are not written at or below a fifth-grade reading level. However, it is clear from this study that it is possible to achieve this level of readability. Increasing the availability and use of plans that meet recognized readability standards may help to improve asthma outcomes, especially in poor populations in which there is both low literacy and the greatest prevalence and severity of asthma. PMID- 11927726 TI - An early (sixth-hour) serum bilirubin measurement is useful in predicting the development of significant hyperbilirubinemia and severe ABO hemolytic disease in a selective high-risk population of newborns with ABO incompatibility. AB - OBJECTIVE: In the era of early discharge of newborns from the hospital, newborns with ABO incompatibility are at especially greater risk for developing a subsequent significant hyperbilirubinemia because some of these infants also may present with some degree of ABO isoimmune disease. In this study, we aimed to determine prospectively the critical serum total bilirubin level to predict significant hyperbilirubinemia and severe hemolytic disease in healthy term newborns with ABO incompatibility based on a serum bilirubin measurement made at a postnatal age at which all newborns are at the hospital before discharge and at which any therapeutic intervention, if necessary, could be started as early as possible. METHODS: A total of 136 healthy term newborns with ABO (O-A or O-B) blood group incompatibility were followed prospectively with daily serum total bilirubin measurements for the first 5 days of life. Newborns with serum total bilirubin levels of > or =5 mg/dL and an increase in serum total bilirubin concentration of >0.5 mg/dL/h in the first 24 hours, > or =12 mg/dL on day 2, > or =15 mg/dL on day 3, and > or =17 mg/dL on days 4 and 5 were defined to have significant hyperbilirubinemia and were started on phototherapy treatment. Additional treatment modalities, including intense phototherapy, intravenous immunoglobulin treatment, and exchange transfusion, were used when serum bilirubin concentrations exceeded 20 mg/dL or increased by >1 mg/dL/h despite a phototherapy treatment of at least 4 hours. The additional assessment of the predictive ability of the sixth-hour serum total bilirubin value in determining the development of significant hyperbilirubinemia was made on the basis of the placement of any of the first 5 days' serum bilirubin measurements in the > or =90th percentile of the study population. On the basis of the percentile tracks constructed from the 10th, 35th, 50th, 60th, and 90th percentiles of serum total bilirubin values, a nomogram demonstrating the 3 percentile tracks as risk zone demarcators with divided risk zones was produced. RESULTS: Twenty-nine newborns (21.3%) had significant hyperbilirubinemia. There were significant differences between the newborns who did and the newborns who did not develop significant hyperbilirubinemia with respect to the reticulocyte count (4.39 +/- 3.46% vs 2.95 +/- 1.63) and the presence of a direct antiglobulin test positivity (6 of 23 vs 0 of 107) and a sibling with neonatal jaundice (6 of 23 vs 5 of 102). A mean serum bilirubin level of > or =4 mg/dL at the sixth hour of life was determined to have the highest sensitivity (86.2%) and negative predictive value (94.5%) and a positive predictive value of 39.7% to predict the newborns who would develop significant hyperbilirubinemia. At the mean serum bilirubin level of 6 mg/dL, the sensitivity, specificity, and negative and positive predictive values were 100%, 91.5%, 100%, and 35.3%, respectively, in diagnosing 6 cases of severe ABO hemolytic disease. On the hour (age)-specific percentile-based nomogram, the zone above the 90th percentile was determined as high risk and that below the 35th percentile as low risk. CONCLUSIONS: The reticulocyte count, a positive direct antiglobulin test, and the presence of a sibling with neonatal jaundice were determined to be the good predictors for the development of significant hyperbilirubinemia and severe hemolytic disease of the newborn. A serum bilirubin measurement and the use of the critical bilirubin levels of 4 mg/dL and 6 mg/dL at the sixth hour of life will predict nearly all newborns who will have significant hyperbilirubinemia and those who will develop severe hemolytic disease of the newborn, respectively. An hour (age)-specific percentile-based nomogram can be used to predict which newborn is at high risk (> or =90th percentile), intermediate risk (35th-90th percentiles), and low risk (<35th percentile) for developing significant hyperbilirubinemia. The 35th and 90th percentile tracks, approximating the serum bilirubin levels of 3.3 mg/dL and 6.5 mg/dL at the sixth hour of life, respectively, can be used as safe risk demarcators in deciding about the time of discharge of ABO-incompatible newborns from the hospital. PMID- 11927727 TI - Continuity of care is associated with high-quality careby parental report. AB - CONTEXT: The benefits of continuity of pediatric care remain controversial. OBJECTIVE: To determine whether there is an association between having a continuous relationship with a primary care pediatric provider and improved quality of care by parental report. DESIGN: Cross-sectional study. SETTING AND POPULATION: Seven hundred fifty-nine patients presenting to a primary care clinic completed surveys, which included validated measures of provider and clinic quality of care from the Consumer Assessment of Health Plan Survey. MAIN EXPOSURE VARIABLE: A continuity of care index that quantifies the degree to which a patient has experienced continuous care with a provider. MAIN OUTCOME MEASURES: The likelihood of parents reporting quality of care as high in several provider- specific items including reporting that providers respected what they had to say, treated them with courtesy and respect, listened to them carefully, explained things in a way they could understand, and spent enough time with their children. In addition, participants were asked to rate the overall quality of the clinic and their child's provider on a 10-point scale. RESULTS: In ordered logistic regression models, continuity of care was associated with statistically significantly higher Consumer Assessment of Health Plan Survey scores for 5 of the 6 items, including feeling that providers respected what parents had to say; listened carefully to them; explained things in a way that they could understand; asked about how their child was feeling, growing, and behaving; and spent enough time with their child. In addition, greater continuity of care was associated with a higher clinic rating, as well as a higher provider rating. CONCLUSIONS: Greater continuity of primary care is associated with higher quality of care as reported by parents. Efforts to improve and maintain continuity may be warranted. PMID- 11927728 TI - Growth and biochemical markers of growth in children with snoring and obstructive sleep apnea. AB - OBJECTIVE: The pathophysiological mechanisms of growth impairment frequently associated with the obstructive sleep apnea syndrome (OSAS) in children are poorly defined. The main objective of this study was to evaluate whether nighttime upper airway obstruction attributable to adenotonsillar hypertrophy and subsequent surgical treatment affect the circulating concentrations of insulin like growth factor-I (IGF-I) and IGF-binding protein 3 (IGFBP-3) along with other growth parameters in children. PATIENTS AND METHODS: We initially studied 70 children (mean age: 5.8 years; range: 2.4-10.5 years) admitted to a university hospital because of clinical symptoms of OSAS. Their sleep was monitored with a 6 channel computerized polygraph. Data on anthropometry and circulating concentrations of IGF-I and IGFBP-3 were generated and compared with corresponding characteristics in control children (N = 35). Thirty children with an obstructive apnea-hypopnea index (OAHI) of 1 or more were categorized as children with OSAS (mean OAHI: 5.4 [95% confidence interval for mean (CI): 3.8 6.9]), whereas 40 children with an OAHI of <1 were considered as primary snorers (PS) (mean OAHI 0.13 [95% CI: 0.05-0.21]). Nineteen children with OAHI >2 underwent adenotonsillectomy attributable to OSAS and were reassessed 6 months later together with 34 nonoperated children with OAHI <2. RESULTS: There were no initial differences in relative height and weight for height between the 3 groups of children. No differences were observed in peripheral IGF-I concentrations, but both OSAS and PS children had reduced peripheral IGFBP-3 levels. The operated children with initial OSAS experienced a highly significant reduction in their OAHI from 7.1 (95% CI: 5.1-9.1) to 0.37 (95% CI: 0.2-0.95). Weight-for-height, body mass index, body fat mass, and fat-free mass increased during the follow-up in the operated children with OSAS, whereas only fat-free mass and relative height increased in the PS children. Both the IGF-I and the IGFBP-3 concentrations increased significantly in the operated children, whereas no significant changes were seen in the PS children. CONCLUSIONS: These observations indicate that growth hormone secretion is impaired in children with OSAS and PS. Respiratory improvement after adenotonsillectomy in children with OSAS results in weight gain and restored growth hormone secretion. PMID- 11927729 TI - Postpartum consequences of an overlap of breastfeeding and pregnancy: reduced breast milk intake and growth during early infancy. AB - OBJECTIVE: Despite cultural pressure to wean when a new pregnancy occurs, some women choose to continue breastfeeding. We determined the effect of an overlap of lactation and late pregnancy on breastfeeding and growth in early infancy. METHODS: We studied 133 Peruvian pregnant women who were > or =18 years of age, had a child <4 years old, and who then had a vaginal birth with a healthy, normal weight infant. Of the 133 women, 68 breastfed during the last trimester of pregnancy (BFP), and 65 had not breastfed during pregnancy (NBFP). On day 2 and at 1-month postpartum, 24-hour intake of breast milk and other liquids was measured. Twice weekly home surveillance documented infant morbidity and dietary intakes. Anthropometry was taken at birth and at 1 month. Maternal anthropometric, health, and socioeconomic status data were collected pre- and postpartum. RESULTS: Pregnant BFP mothers breastfed 5.3 +/- 4.3 times/day. BFP and NBFP infants did not differ in breastfeeding behavior or in colostrum intake on day 2. BFP infants breastfed longer per feed and per 24 hours (35.2 minutes/24 hours) than did NBFP infants; however, 1-month intakes per feed tended to be lower among the BFP infants. After controlling for confounders, BFP infants gained 125 g less than did NBFP infants (about 15% of mean weight gain). A sustained decline would result in a -0.7 z score change in weight-for-age by 6 months. CONCLUSIONS: A lactation-pregnancy overlap had a negative effect on early infant outcomes. Additional studies are needed to determine whether the effect continues past 1 month of age. PMID- 11927730 TI - Correlates of lactation in mothers of very low birth weight infants. AB - OBJECTIVE: We sought to determine the correlates of intent to breastfeed and of successful lactation and nursing at the breast in mothers of very low birth weight (VLBW; <1.5 kg) infants. METHODS: We conducted a prospective observational study of 119 mothers of singleton VLBW infants (mean birth weight: 1056 g; mean gestational age: 28 weeks), 87 (73%) of whom intended to breastfeed. Mothers completed questionnaires at 3 weeks' postnatal age and at 35 and 40 weeks' (term) and 4 months' corrected ages (CAs). RESULTS: Of the 87 mothers who intended to breastfeed, 30 mothers (34%) continued lactation beyond 40 weeks' CA (postmenstrual plus postnatal age). Compared with mothers who discontinued lactation before this time, those who continued were older (31 vs 26 years), more were married (80% vs 39%), they had more than a high school education (70% vs 42%), and they were white (63% vs 35%). There were no significant differences in their infants' birth data or rates of neonatal morbidity. Significant correlates of lactation beyond 40 weeks' CA included beginning milk expression before 6 hours postdelivery, expressing milk > or =5 times per day, and kangaroo care. These correlates remained significant after controlling for maternal age, race, marital status, and education beyond high school. At 4 months' CA, 14 mothers (16%) were still lactating, 12 of whom were nursing at the breast. CONCLUSIONS: Increased maternal support specifically directed toward behavioral factors, including early and more frequent milk expression and kangaroo care, may improve the rates of successful lactation among mothers of VLBW infants who choose to breastfeed. PMID- 11927732 TI - Economic evaluation of different methods of screening for amblyopia in kindergarten. AB - OBJECTIVE: To compare the cost-effectiveness of 5 methods of screening for untreated amblyopia in kindergarten from a third-party-payer perspective: A) uncorrected monocular visual acuity testing with pass threshold > or =0.5 (20/40) and < or =1 line difference between eyes; B) same as A, but pass threshold > or =0.6 (20/32); C) same as A, plus cover tests and examination of eye motility and head posture; D) same as C, but pass threshold > or =0.6 (20/32); and E) refractive screening without cycloplegia using the Nikon Retinomax autorefractor. METHODS: A decision-analytic model was used with a time horizon until diagnostic examination. According to the model, all 3-year-old children were screened in kindergarten with 1 of the screening methods. Children with positive screening results were referred to an ophthalmologist for diagnostic examination. Children with inconclusive screening results were either referred to an ophthalmologist directly (option 1) or rescreened by the same method after 1 year and referred to an ophthalmologist if rescreening was positive or inconclusive (option 2). Screening test characteristics and costs were estimated on the basis of a field study in which 1180 3-year-old children were examined by orthoptists in 121 German kindergartens. RESULTS: Compared with methods A option 1 (A-1), B-1, C-1, C-2, E-1, and E-2, there was at least 1 other method that was both less costly and more effective. The average costs per detected case were lowest for method A 2 (878 Euro), followed by methods B-2 (886 Euro), D-2 (908 Euro), and D-1 (965 Euro). When these methods were compared with each other, the additional costs per extra case detected were 1058 Euro (B-2 vs A-2), 1359 Euro (D-2 vs B-2), and 13 448 Euro (D-1 vs D-2). CONCLUSIONS: Monocular visual acuity screening with rescreening of inconclusive results had a favorable cost-effectiveness. By adding additional test items, few more cases could be detected. Because of a great proportion of false-negative, false-positive, and inconclusive results, refractive screening was less effective with an unfavorable cost-effectiveness. PMID- 11927731 TI - Safety and reactogenicity of a novel DTPa-HBV-IPV combined vaccine given along with commercial Hib vaccines in comparison with separate concomitant administration of DTPa, Hib, and OPV vaccines in infants. AB - OBJECTIVE: Combination vaccines simplify vaccine administration and have the potential to promote compliance and cost-effectiveness by decreasing the number of injections needed to immunize a child. The objective of this study was to assess the safety and reactogenicity of the diphtheria-tetanus toxoid-acellular pertussis-hepatitis B virus-inactivated polio virus (DTPa-HBV-IPV) vaccine when coadministered with different Haemophilus influenzae type B (Hib) vaccines in comparison with separate, commercially available, control vaccines in a 3-dose primary vaccination series. METHODS: An open-label, randomized, parallel-group study in 5318 infants who were 8 to 16 weeks of age at enrollment was conducted in 90 centers in Germany. The incidence of adverse events that occurred in infants who received the DTPa-HBV-IPV candidate vaccine coadministered with 1 of 4 different Hib vaccines (given in separate sites; groups 1-4) was compared with the incidence that occurred in infants who received commercially available control vaccines (DTPa, Hib, and oral polio virus [OPV] vaccine; group 5) administered separately. The vaccines were given as a 3-dose primary series at 3, 4, and 5 months of age. Infants were assessed for solicited local and general adverse events for 4 days and for unsolicited adverse events for 30 days after each vaccine dose. The primary endpoint was to rule out a 7.5% increase in infants who experienced grade 3 (defined as preventing normal everyday activities unless otherwise specified) solicited local and general adverse events over the 3 dose primary course after the combined DTPa-HBV-IPV vaccine coadministered with Hib as compared with commercially available vaccines. RESULTS: During the 3-dose primary course, 490 of 3029 infants (16.2%) in the pooled DTPa-HBV-IPV vaccine groups and 151 of 744 (20.3%) in the control vaccine group experienced a grade 3 adverse event (rate difference [control minus combination] 4.1%; 90% confidence interval, 1.41-7.13). The lower limit of the 90% confidence interval of the observed difference remained above the prespecified -7.5% limit for noninferiority, thereby meeting the primary endpoint. The incidences of local injection-site reactions were similar for the DTPa-HBV-IPV and DTPa injection sites. Significant differences in the incidence of both local and general adverse events were observed depending on which of the Hib vaccines was coadministered. Infants who received Hib N meningitidis outer-membrane complex protein conjugate vaccine had greater incidences of fever and, to a lesser extent, greater reactions at the Hib injection site than did infants who received other Hib vaccines. CONCLUSIONS: The combination DTPa-HBV-IPV vaccine administered concomitantly with Hib vaccine at separate sites was at least as safe as coadministration of individual DTPa, Hib, and OPV vaccines in terms of the defined endpoints for safety. PMID- 11927733 TI - Zidovudine and perinatal human immunodeficiency virus type 1 transmission: a population-based approach. AB - OBJECTIVE: This study examined the impact of the full 3-arm zidovudine regimen on the perinatal transmission of human immunodeficiency virus type 1 (HIV-1) using population-based data. METHODS: We retrospectively ascertained information on zidovudine prescription and other characteristics of HIV-infected pregnant women and children for birth cohort years 1993, 1995, 1996, and 1997 using HIV/acquired immunodeficiency syndrome registry data from a state health department supplemented by medical record reviews. RESULTS: The transmission rate decreased from 12.5% in 1993 to 4.6% in 1997. The proportions of HIV-1-infected mothers and children who were prescribed all 3 arms of zidovudine increased from 68% in 1995 to 93% in 1997. Unadjusted and adjusted odds ratios for the relationship between the prescription of 3 arms of zidovudine and the infants' HIV status were 0.19 (95% confidence interval: 0.05-0.84) and 0.15 (95% confidence interval: 0.02 0.96), respectively. CONCLUSION: Perinatal HIV-1 transmission rates have decreased over time. This study demonstrates the effectiveness of the rapid implementation of the United States Public Health Service recommendations for the comprehensive use of zidovudine among HIV-1-infected pregnant women in a predominantly rural state. PMID- 11927734 TI - Reported adherence as a determinant of response to highly active antiretroviral therapy in children who have human immunodeficiency virus infection. AB - OBJECTIVE: The complexity of highly active antiretroviral therapy (HAART), with multiple medications, formulations, and dosing intervals, makes adherence challenging. Little is known about the adherence of children to HAART. The objective of this study was to identify correlates of adherence to HAART and the relationship between adherence and study outcomes in a pediatric clinical trial. METHODS: Pediatric AIDS Clinical Trials Group 377 is a phase I/II randomized trial of 4 HAART regimens in antiretroviral-experienced, clinically stable children aged 4 months to 17 years. The 4 treatment arms include various 3- or 4 drug combinations of d4T, 3TC, nevirapine, ritonavir, and nelfinavir. After informed consent was obtained, 193 children were enrolled between December 1997 and September 1998. Questionnaires were developed to collect subject- or caregiver-reported adherence to study medications and to identify problems associated with medication administration. Every 3 months, the number of doses of each medication missed during the 3 days preceding the study visit was recorded. Full adherence (FA) and non-full adherence were defined as missing no doses and missing at least 1 dose, respectively. RESULTS: Adherence data from study week 48 or the most recent study visit were available for 125 children (week 48 for 109 children). Overall, 70% of children reported FA and 30% reported non-full adherence. Adherence did not differ by treatment arm, age, or the child's knowledge of his or her human immunodeficiency virus infection status. There was a suggestion that adherence was less for white than nonwhite children (40% vs 73% FA) and did not differ between black and Hispanic children. Rates of FA were 82% for d4T, 79% for 3TC, 83% for nevirapine, 84% for ritonavir, and 68% for nelfinavir. Despite the similar rates of FA, difficulties with taking specific medications were reported most frequently for ritonavir and nelfinavir. These included poor taste, patient refusal, and scheduling problems. Adherence was associated with the virologic response: FA was seen in 92% of children with > or =2 log10 drop in viral load and in 64% with <2 log10 drop in viral load. CONCLUSION: In children, reported adherence predicts the virologic response to HAART therapy and is a useful measure of adherence. Interventions and regimens to increase adherence to HAART should result in an improved outcome. PMID- 11927735 TI - Ondansetron decreases vomiting associated with acute gastroenteritis: a randomized, controlled trial. AB - OBJECTIVE: Relatively little research has examined the role of antiemetic agents in the treatment of acute gastroenteritis. The use of the selective 5-HT3 receptor antagonists (eg, ondansetron) offers a potentially valuable treatment option. The objective of this study was to evaluate the efficacy of ondansetron for the treatment of vomiting associated with acute gastroenteritis in children. METHODS: A randomized, double blind, placebo-controlled trial was conducted in the emergency department of a tertiary-care children's hospital. Eligible patients were 1 month to 22 years old and required intravenous fluids for gastroenteritis. Of 172 patients approached, 107 were enrolled (54 to intravenous ondansetron, 53 to placebo). The mean age was 5.3 years, and 53% of the patients were male. The frequency of vomiting, admission rate, and occurrence of complications were measured. RESULTS: After drug administration, 38 (70%) of the 54 patients in the ondansetron group had complete cessation of vomiting compared with 27 (51%) of the 53 patients in the placebo group. Sixteen (30%) of the 53 patients in the placebo group required admission compared with 14 (26%) of the 54 in the ondansetron group. An analysis of previously untreated patients with a measured serum carbon dioxide > or =15 mEq/L showed that 11 (23%) of the 47 who received placebo were admitted compared with 3 (7%) of the 43 who received ondansetron. No significant complications were detected. CONCLUSIONS: Intravenous ondansetron decreases vomiting in children with gastroenteritis. In addition, ondansetron reduces the need for admission in those who are treated at an initial visit to the emergency department and have a measured serum carbon dioxide > or =15 mEq/L. The safety and low cost of this therapy suggests that ondansetron can be valuable in treating gastroenteritis in children. PMID- 11927736 TI - Survival with hypopituitarism from congenital syphilis. AB - Congenital syphilis continues to occur despite the advances in testing of pregnant women in many countries and the availability of penicillin since 1943. This is a report of a child with multiple systemic manifestations of congenital syphilis. This is one of the few cases of survival with pituitary involvement from congenital syphilis and the first noted case in which diabetes insipidus developed. Institution of routine testing in all countries might have prevented the severe manifestations of syphilis seen in this child. PMID- 11927737 TI - Resistance to recombinant human granulocyte colony-stimulating factor in neonatal alloimmune neutropenia associated with anti-human neutrophil antigen-2a (NB1) antibodies. AB - Neonatal alloimmune neutropenia is the neutrophil counterpart of the erythrocyte disorder of hemolytic disease of the newborn. Fetal neutrophil antigens, which are inherited from the father but foreign to the pregnant mother, provoke the formation of maternal antibodies, which, on transplacental passage, cause fetal/neonatal neutropenia. Because infants with this disorder are at a higher risk of infection, recombinant hematopoietic growth factors, such as recombinant human granulocyte colony-stimulating factor, have been tried, with generally good results, to treat those with severe and prolonged neutropenia. We report a neonate who had neonatal alloimmune neutropenia associated with antibodies directed against human neutrophil antigen-2a (NB1) and initially failed to respond to even very high doses of recombinant human granulocyte colony stimulating factor but eventually had a therapeutic response. PMID- 11927738 TI - A rare presentation of childhood pompe disease: cardiac involvement provoked by Epstein-Barr virus infection. AB - Myocarditis attributed to Epstein-Barr virus (EBV) as the sole cause is a rare manifestation. Myocarditis ascribed to EBV infection in combination with other factors has been reported in a few more cases. We report a child who experienced active EBV infection and later, at 19 months of age, received a diagnosis of Pompe disease (acid alpha-glucosidase deficiency) with predominant cardiac involvement. The cardiac symptoms resolved at the end of the EBV infection. When the patient was recently seen, at 8 years of age, she had an increased left ventricular wall thickness but normal cardiac function. DNA analysis identified this patient as compound heterozygote for a mutant Tyr292Cys and a null allele. In light of genotype-phenotype correlation, it is notable that a Spanish patient with a functionally similar genotype (Tyr292Cys/Arg854Stop) also had childhood Pompe disease with peripheral muscular involvement. PMID- 11927739 TI - Peroneal nerve palsy: a complication of umbilical artery catheterization in the full-term newborn of a mother with diabetes. AB - Umbilical artery catheters are an essential aid in the treatment of newborn infants who have cardiopulmonary disease. However, it is well-known that umbilical artery catheterization is associated with complications. The most frequent visible problem in an umbilical line is blanching or cyanosis of part or all of a distal extremity or the buttock area resulting from either vasospasm or a thrombotic or embolic incidence. Ischemic necrosis of the gluteal region is a rare complication of umbilical artery catheterization. We report the case of a full-term infant of an insulin-dependent diabetic mother with poor blood glucose control who developed a left peroneal nerve palsy after ischemic necrosis of the gluteal region after umbilical artery catheterization. The infant was born weighing 5050 g. The mother of the infant had preexisting diabetes mellitus that was treated with insulin from the age of 14 years. The metabolic control of the mother had been unstable both before and during the pregnancy. The neonate developed respiratory distress syndrome soon after birth and was immediately transferred to the neonatal intensive care unit. Mechanical ventilation via endotracheal tube was quickly considered necessary after rapid pulmonary deterioration. Her blood glucose levels were 13 mg/dL. A 3.5-gauge umbilical catheter was inserted into the left umbilical artery for blood sampling without difficulty when the infant required 100% oxygen to maintain satisfactory arterial oxygen pressure. Femoral pulses and circulation in the lower limbs were normal immediately before and after catheterization. A radiograph, which was taken immediately, showed the tip of the catheter to be at a level between the fourth and fifth sacral vertebrae. The catheter was removed immediately. Circulation and femoral pulses were normal and no blanching of the skin was observed. Another catheter was repositioned and the tip was confirmed radiologically to be in the thoracic aorta between the sixth and seventh thoracic vertebrae. The catheter was continuously flushed with heparinized solution. Three days after umbilical arterial catheterization, bruising was observed over the left gluteal region. The catheter was immediately removed despite its correct position. Over the next few days, the bruised skin and underlying tissues became necrotic. The area affected was 3 x 4 cm in diameter, with central necrosis surrounded by a rim of dark, red skin, which, in turn, was sharply demarcated from normal skin by a narrow, pale zone. Surgical excision of the gluteal necrosis was performed, but a deep ulcer 3 cm in diameter was left. The gluteal ulcer required 1 month to heal completely with extensive scar tissue formation. Throughout this period, the infant showed active movements in all of her limbs. At 4 weeks of age deterioration of all movement below the left knee with a dropping foot was observed. Severe peroneal nerve palsy was confirmed through nerve conduction studies, and there was electromyographic evidence of degeneration of the muscles supplied by the peroneal branch of the sciatic nerve. A Doppler study, which was also conducted, revealed no vascular damage. Treatment with physiotherapy and night-splinting of the left ankle was instituted. Repeated examination and nerve conduction tests at 3 months showed slow improvement with the left peroneal nerve remaining nonexcitable. At the time of this writing, the infant is 6 months old, and muscular strength below the left knee is still weak and atrophic changes in the form of muscle-wasting are already present. The rest of her motor development is normal. In our case, gangrene of the buttocks and sciatic nerve palsy followed displacement of the tip of the catheter into the inferior gluteal artery, a main branch of the internal iliac artery supplying the gluteus maximus, the overlying skin, and the sciatic nerve. The gangrenous changes were probably caused by vascular occlusion resulting from catheter-induced vasospasm of the inferior gluteal artery. Sciatic nerve palsy associated with umbilical artery catheterization has been postulated to be caused by vascular occlusion of the inferior gluteal artery. Infants of diabetic mothers may exhibit changes in coagulation factors and be at increased risk of thrombotic complications in utero and postnatally. In addition, maternal diabetes mellitus is associated with an increased incidence of congenital abnormalities, the incidence of which is 3 to 5 times higher than that among nondiabetic mothers. Although no particular or specific abnormalities have been associated with maternal disabilities, abnormalities of the cardiovascular system, including the development of umbilical vessels, frequently occur. This complication of umbilical artery catheterization has not been widely reported. We describe the first case that refers to gluteal gangrene and peroneal nerve palsy after umbilical artery catheterization of a newborn infant of a diabetic mother with poor blood glucose control. It should be noted that there were no contributing factors except that of the displacement of the catheter into the inferior gluteal artery. We speculate that the displacement of the tip of the catheter, with no difficulty in the present case, was associated with the maldevelopment of normal branching patterns of arteries after exposure of the fetus to hyperglycemia. In conclusion, umbilical artery catheterization is possibly associated with vascular occlusion, particularly in infants of diabetic mothers. Frequent inspection after the procedure has been performed is of the utmost importance especially in these neonates who often suffer from cardiopulmonary disease and require catheterization of their umbilical artery. PMID- 11927740 TI - Treatment of plastic bronchitis in a Fontan patient with tissue plasminogen activator: a case report and review of the literature. AB - Plastic bronchitis is a condition in which large, bronchial casts with rubber like consistency develop in the tracheobronchial tree and cause airway obstruction. We describe a 4-year-old girl who had Fontan physiology and who developed plastic bronchitis and report for the first time the use of aerosolized tissue plasminogen activator for treatment of this condition. The literature is reviewed with emphasis placed on the occurrence of this disorder in patients with single ventricle physiology. PMID- 11927741 TI - The psychological maltreatment of children--technical report. AB - Psychological maltreatment is a common consequence of physical and sexual abuse but also may occur as a distinct entity. Until recently, there has been controversy regarding the definition and consequences of psychological maltreatment. Sufficient research and consensus now exist about the incidence, definition, risk factors, and consequences of psychological maltreatment to bring this form of child maltreatment to the attention of pediatricians. This technical report provides practicing pediatricians with definitions and risk factors for psychological maltreatment and details how pediatricians can prevent, recognize, and report psychological maltreatment. Contemporary references and resources are provided for pediatricians and parents PMID- 11927742 TI - Technical report: diagnosis and management of childhood obstructive sleep apnea syndrome. AB - OBJECTIVE: This technical report describes the procedures involved in developing the recommendations of the Subcommittee on Obstructive Sleep Apnea Syndrome in children. The group of primary interest for this report was otherwise healthy children older than 1 year who might have adenotonsillar hypertrophy or obesity as underlying risk factors of obstructive sleep apnea syndrome (OSAS). The goals of the committee were to enhance the primary care clinician's ability to recognize OSAS, identify the most appropriate procedure for diagnosis of OSAS, identify risks associated with pediatric OSAS, and evaluate management options for OSAS. METHODS: A literature search was initially conducted for the years 1966 1999 and then updated to include 2000. The search was limited to English language literature concerning children older than 2 and younger than 18 years. Titles and abstracts were reviewed for relevance, and committee members reviewed in detail any possibly appropriate articles to determine eligibility for inclusion. Additional articles were obtained by a review of literature and committee members' files. Committee members compiled evidence tables and met to review and discuss the literature that was collected. RESULTS: A total of 2115 titles were reviewed, of which 113 provided relevant original data for analysis. These articles were mainly case series and cross-sectional studies; overall, very few methodologically strong cohort studies or randomized, controlled trials concerning OSAS have been published. In addition, a minority of studies satisfactorily differentiated primary snoring from true OSAS. Reports of the prevalence of habitual snoring in children ranged from 3.2% to 12.1%, and estimates of OSAS ranged from 0.7% to 10.3%; these studies were too heterogeneous for data pooling. Children with sleep-disordered breathing are at increased risk for hyperactivity and learning problems. The combined odds ratio for neurobehavioral abnormalities in snoring children compared with controls is 2.93 (95% confidence interval: 2.23-3.83). A number of case series have documented decreased somatic growth in children with OSAS; right ventricular dysfunction and systemic hypertension also have been reported in children with OSAS. However, the risk growth and cardiovascular problems cannot be quantified from the published literature. Overnight polysomnography (PSG) is recognized as the gold standard for diagnosis of OSAS, and there are currently no satisfactory alternatives. The diagnostic accuracy of symptom questionnaires and other purely clinical approaches is low. Pulse oximetry appears to be specific but insensitive. Other methods, including audiotaping or videotaping and nap or home overnight PSG, remain investigational. Adenotonsillectomy is curative in 75% to 100% of children with OSAS, including those who are obese. Up to 27% of children undergoing adenotonsillectomy for OSAS have postoperative respiratory complications, but estimates are varied. Risk factors for persistent OSAS after adenotonsillectomy include continued snoring and a high apnea-hypopnea index on the preoperative PSG. CONCLUSIONS: OSAS is common in children and is associated with significant sequelae. Overnight PSG is currently the only reliable diagnostic modality that can differentiate OSAS from primary snoring. However, the PSG criteria for OSAS have not been definitively validated, and it is not clear that primary snoring without PSG-defined OSAS is benign. Adenotonsillectomy is the first-line treatment for OSAS but requires careful postoperative monitoring because of the high risk of respiratory complications. Adenotonsillectomy is usually curative, but children with persistent snoring (and perhaps with severely abnormal preoperative PSG results) should have PSG repeated postoperatively. PMID- 11927743 TI - Managing anemia in a pediatric office practice: Part 2. PMID- 11927744 TI - Management of substance abuse. PMID- 11927745 TI - Pneumonia. PMID- 11927746 TI - Index of suspicion. PMID- 11927747 TI - Corticosteroids. PMID- 11927748 TI - Unveiling the secrecy behind masturbation. PMID- 11927749 TI - Clindamycin. PMID- 11927750 TI - Serologic testing. PMID- 11927751 TI - Somatization: a critical review of conceptual and methodological issues. AB - Despite its apparent clinical importance and the extensive research that has been conducted in the past decades, somatization remains a complex concept. Two distinct ways of looking at somatization can be distinguished: somatization as a phenomenon that is secondary to psychological distress (presenting somatization) and somatization as a primary phenomenon characterized by medically unexplained symptoms (functional somatization). The literature was analyzed in terms of this distinction and a selective review was conducted, focusing on a critical analysis of conceptual and methodological issues related to presenting and functional somatization. A number of measurement issues related to somatization in general were also highlighted. On the basis of the available data, the strengths and weaknesses of different concepts are pointed out. Conclusions are formulated regarding which concepts or approaches might be useful both clinically and from a research perspective. Finally, a number of suggestions for future research are offered. PMID- 11927752 TI - The prevalence of distress in persons with human immunodeficiency virus infection. AB - The purpose of this study was to assess the prevalence of distress, anxiety, and depression in persons with human immunodeficiency virus (HIV) infection and determine the feasibility of screening in an urban HIV primary care setting. A convenience sample of 101 patients in the waiting room of an acquired immunodeficiency syndrome clinic completed two questionnaires, the Hospital Anxiety and Depression Scale (HADS) and the Distress Thermometer. The patient's demographic, medical, and psychiatric histories were obtained through chart review. The results of the Distress Thermometer revealed that 72.3% had a score of 5 or greater, demonstrating high distress. The results of the HADS revealed that 70.3% had high anxiety, with a score of 7 or greater. On the HADS depression questions, 45.5% had a score of 7 or greater, indicating depression. Analysis of the total HADS scores, including anxiety and depression, revealed that 53.5% had a score of greater than 15 and were experiencing significant distress. Patients with high viral loads were more likely to be distressed (P < 0.0005). Patients with high viral loads were also more likely to have higher anxiety or depression scores on the HADS. Patients who had CD4 counts higher than 500/mm(3) were less likely to be depressed. This study demonstrates a high prevalence of distress, anxiety, and depression among persons with HIV. The HADS and the Distress Thermometer showed a good correlation with each other (P < 0.0005), and these questionnaires can provide a simple and efficient method for rapid screening in an HIV clinic setting. PMID- 11927753 TI - Psychosocial factors associated with quality of life in Japanese patients with human immunodeficiency virus infection. AB - The purpose of this study was to identify psychosocial factors, including coping style, that are associated with quality of life (QOL) in Japanese patients with human immunodeficiency virus (HIV) infection and to clarify the difference in coping style between patients with hemophilia and patients with sexually transmitted infection. We administered the World Health Organization QOL-26 questionnaire, the Mental Adjustment Toward Cancer scale, and the Social Support scale to 50 patients with HIV infection. Regarding QOL scores, Fighting Spirit was the positive coping style; Helplessness/Hopelessness and Anxious Preoccupation were negative coping styles. Psychological QOL scores in patients with hemophilia were lower than those in patients with sexually transmitted infection. Patients with hemophilia had a significantly weaker Fighting Spirit than patients with sexually transmitted infection. PMID- 11927754 TI - Medical comorbidity in psychiatric inpatients: relation to clinical outcomes and hospital length of stay. AB - Medical comorbidity is common in psychiatric inpatients and may be associated with substantial impairment and mortality. Few studies have examined the relation between this comorbidity and psychiatric outcomes. A series of 950 admissions to the Johns Hopkins Hospital Phipps Psychiatric Service were rated by attending psychiatrists at admission and discharge on symptom and functional measures. A subset was also evaluated on the General Medical Health Rating, a valid and reliable measure of seriousness of medical comorbidity. Attending psychiatrists were also asked at discharge whether medical comorbidity had been a focus of care during the hospitalization; medical comorbidity had been a focus of care in about 20% of the patients. Serious active medical comorbidity was present in 15% of patients on admission and 12% at discharge. Medical comorbidity was associated with a 10%-15% increase in psychiatric symptoms and functional impairment at discharge, even after adjustment for admission clinical status. In addition, when comorbidity had been a focus of care during the hospitalization, length of stay was prolonged by 3.25 days on average. Medical comorbidity has measurable effects on the psychiatric outcomes of psychiatric inpatients and in some cases prolongs hospital stay. Psychiatrists should redouble their efforts to detect and treat this comorbidity and should consider whether special inpatient units might be needed to care for psychiatric patients with complex medical comorbidity. PMID- 11927755 TI - Tricyclic antidepressants and cognitive decline. AB - In this study, we sought to determine the cross-sectional and longitudinal relations of tricyclic antidepressant (TCA) use to cognitive function and cognitive change in a population-based sample of adults (n=1,488). Sociodemographic information, TCA use, and baseline scores on the Mini-Mental State Exam (MMSE) were determined in the initial two waves of the study. At wave 3, participants repeated the MMSE; the prospective relation was assessed for change between waves 2 and 3 (median 11.5 years). These findings failed to support the concept that TCA use is related to concurrent measurable cognitive deficits, and TCA use does not appear to significantly compromise memory over a substantial time span. PMID- 11927756 TI - Clinical outcomes following a trial of sertraline in rheumatoid arthritis. AB - We report an open-label trial of sertraline in the treatment of major depression in 54 consecutive rheumatoid arthritis (RA) patients meeting DSM-IV criteria for major depressive disorder. We initially surveyed 628 RA outpatients with the Center for Epidemiologic Studies Depression Scale (CES-D) and invited those with depression to be evaluated further and treated. Eighty-four RA patients reporting depressive symptoms agreed to participate in person, and 56 met the criteria for major depressive disorder. Of these 56 patients, 54 agreed to medication treatment and were enrolled in the study. Patients were also randomized to one of three psychological treatment conditions, but for this study, conditions were collapsed because previous research on this sample indicated no significant between-group differences in depression after treatment. Patients were assessed with the CES-D and the Hamilton Rating Scale for Depression after the intervention, at 6-month follow-up, and at 15-month follow-up. At the last follow up, 41 patients remained for assessment. In this study, sertraline was found to be a safe and efficacious treatment of depression complicating RA. PMID- 11927757 TI - Psychological correlates of pain behavior in patients with chronic low back pain. AB - Pain behaviors that are excessive for the degree of known physical disease are common in patients with chronic low back pain and are frequently assumed to arise from a comorbid depressive illness. Although some studies have confirmed an association between depression and excessive pain behavior, methodologic problems (such as the use of depression ratings that also recorded symptoms attributable to physical disease) make interpretation of this finding difficult. We recruited 54 consecutive patients with chronic (>6 months) low back pain from a hospital clinic. Subjects completed self-rated assessments of anxiety and depression (Hospital Anxiety and Depression Scale) designed to be minimally affected by physical symptoms, along with assessments of disability (ODQ), pain (visual analogue scale), pain behavior (Waddell checklist), and physical impairment. Seventeen subjects (31%) exhibited excessive pain behavior. Overall, they were no more depressed or anxious than the remainder, although men with excessive pain behavior showed a trend toward being more depressed. Patients with excessive pain behavior were more disabled (self-rated and observer-rated), reported greater pain, and were more likely to be female and to have pain of shorter duration. Pain behavior did not correlate with anxiety or depression, but correlated with measures of disability and pain intensity. Factor analysis revealed that physical disability, pain intensity, and pain behavior loaded heavily on the first factor. Anxiety and depression loaded together on a separate factor. We conclude that pain behaviors were not related to anxiety or depression in our group, although gender differences between groups could have contributed to our negative findings. Pain behaviors may influence other physical measures. Further studies are required to investigate the relation between depression and pain behavior while controlling for gender differences. PMID- 11927758 TI - Association between pretransplant psychological assessments and posttransplant psychiatric disorders in living-related transplantation. AB - The authors examined pretransplant assessment in order to predict posttransplant occurrence of psychiatric disorders in living-related transplantation (LRT). Before LRT, the authors administered the Integrated House-Tree-Person Drawing Test (I-HTP) and 20-item Toronto Alexithymia Scale (TAS-20) to 31 donor-recipient pairs undergoing living-related liver transplantation (LRLT) and 65 pairs undergoing living-related kidney transplantation (LRKT). After LRT, the authors examined the occurrence of psychiatric disorders for the recipients and donors. Pretransplant, two psychological indicators,-alexithymia, a lack of verbalized emotion and abnormal projective drawings such as truncated tree representation were significantly related to the manifestation of paradoxical psychiatric syndrome (PPS) in LRLT and LRKT. The occurrence of PPS was significantly related to recipients' guilt feelings toward living donors, but these were strongly superseded by recipients' desires to escape from approaching death just before LRT. These results suggest that pretransplant psychological assessment is useful for predicting posttransplant occurrence of psychiatric disorders. PMID- 11927759 TI - Relationship between somatosensory amplification and alexithymia in a Japanese psychosomatic clinic. AB - To examine the relationship between somatosensory amplification and three factors of alexithymia (difficulty identifying feelings, difficulty describing feelings, and externally oriented thinking), 48 outpatients attending a Japanese psychosomatic clinic and 33 comparative outpatients completed the Somatosensory Amplification Scale (SSAS), 20-item Toronto Alexithymia Scale (TAS-20), Profile of Mood States (POMS), and other self-rating questionnaires. The scores on the SSAS and the first and second TAS-20 factors were higher (all P<0.001) in the psychosomatic group than in the comparison group. The SSAS was positively associated (both P<0.01) with these two TAS-20 factors, controlling for the effects of age, sex, group, and POMS tension-anxiety and depression. Somatosensory amplification appears to be associated with difficulties identifying and describing feelings, not externally oriented thinking, in Japanese patients. PMID- 11927761 TI - Intractable delirium associated with ziconotide successfully treated with electroconvulsive therapy. PMID- 11927760 TI - Ziprasidone treatment of delirium. PMID- 11927762 TI - Recovery from new-onset diabetes in a schizophrenic man after withdrawal of olanzapine. PMID- 11927763 TI - Death from clozapine-induced constipation: case report and literature review. PMID- 11927764 TI - Acute stress disorder following ventilation. PMID- 11927765 TI - Med-psych drug-drug interactions update. PMID- 11927767 TI - Case of a 73-year-old man with dementia and a likely pheochromocytoma mistaken for an anxiety disorder. PMID- 11927766 TI - Psychiatric symptoms in a case of intracranial tuberculosis. PMID- 11927769 TI - Overemphasis of psychological risks of genetic testing may have "dire" consequences. PMID- 11927768 TI - Probable Lewy body dementia presenting as "delirium". PMID- 11927770 TI - Childhood trauma and somatic preoccupation in adulthood among a sample of psychiatric inpatients. PMID- 11927771 TI - Use and citation of Beck Depression Inventory to assess depression in HIV infection. PMID- 11927772 TI - SWORDS: a statistical tool for analysing large DNA sequences. AB - In this article, we present some simple yet effective statistical techniques for analysing and comparing large DNA sequences. These techniques are based on frequency distributions of DNA words in a large sequence, and have been packaged into a software called SWORDS. Using sequences available in public domain databases housed in the Internet, we demonstrate how SWORDS can be conveniently used by molecular biologists and geneticists to unmask biologically important features hidden in large sequences and assess their statistical significance. PMID- 11927773 TI - Ab initio gene identification: prokaryote genome annotation with GeneScan and GLIMMER. AB - We compare the annotation of three complete genomes using the ab initio methods of gene identification GeneScan and GLIMMER. The annotation given in GenBank, the standard against which these are compared, has been made using GeneMark. We find a number of novel genes which are predicted by both methods used here, as well as a number of genes that are predicted by GeneMark, but are not identified by either of the nonconsensus methods that we have used. The three organisms studied here are all prokaryotic species with fairly compact genomes. The Fourier measure forms the basis for an efficient non-consensus method for gene prediction, and the algorithm GeneScan exploits this measure. We have bench-marked this program as well as GLIMMER using 3 complete prokaryotic genomes. An effort has also been made to study the limitations of these techniques for complete genome analysis. GeneScan and GLIMMER are of comparable accuracy insofar as gene-identification is concerned, with sensitivities and specificities typically greater than 0.9. The number of false predictions (both positive and negative) is higher for GeneScan as compared to GLIMMER, but in a significant number of cases, similar results are provided by the two techniques. This suggests that there could be some as-yet unidentified additional genes in these three genomes, and also that some of the putative identifications made hitherto might require re-evaluation. All these cases are discussed in detail. PMID- 11927774 TI - Comparative genomics using data mining tools. AB - We have analysed the genomes of representatives of three kingdoms of life, namely, archaea, eubacteria and eukaryota using data mining tools based on compositional analyses of the protein sequences. The representatives chosen in this analysis were Methanococcus jannaschii, Haemophilus influenzae and Saccharomyces cerevisiae. We have identified the common and different features between the three genomes in the protein evolution patterns. M. jannaschii has been seen to have a greater number of proteins with more charged amino acids whereas S. cerevisiae has been observed to have a greater number of hydrophilic proteins. Despite the differences in intrinsic compositional characteristics between the proteins from the different genomes we have also identified certain common characteristics. We have carried out exploratory Principal Component Analysis of the multivariate data on the proteins of each organism in an effort to classify the proteins into clusters. Interestingly, we found that most of the proteins in each organism cluster closely together, but there are a few 'outliers'. We focus on the outliers for the functional investigations, which may aid in revealing any unique features of the biology of the respective organisms PMID- 11927775 TI - Horizontal gene transfer and bacterial diversity. AB - Bacterial genomes are extremely dynamic and mosaic in nature. A substantial amount of genetic information is inserted into or deleted from such genomes through the process of horizontal transfer. Through the introduction of novel physiological traits from distantly related organisms, horizontal gene transfer often causes drastic changes in the ecological and pathogenic character of bacterial species and thereby promotes microbial diversification and speciation. This review discusses how the recent influx of complete chromosomal sequences of various microorganisms has allowed for a quantitative assessment of the scope, rate and impact of horizontally transmitted information on microbial evolution. PMID- 11927778 TI - Variations in angiotensin-converting enzyme gene insertion/deletion polymorphism in Indian populations of different ethnic origins. AB - The pattern of angiotensin-converting enzyme (ACE) gene insertion/deletion (I/D) polymorphism in the Indian population is poorly known. In order to determine the status of the polymorphism, young unrelated male army recruits were screened. The population had cultural and linguistic differences and lived in an environment that varied significantly from one region to another. Analysis of the genotype, showed higher frequency of the insertion allele in four of the five groups i.e. I allele frequency was significantly higher (P < 0.05) in Dogras, Assamese and Kumaonese. The deletion allele frequency was comparatively higher in the fifth group that belonged to Punjab. A correlation was observed between the genotype and enzyme activity. Involvement of a single D allele in the genotype enhanced the activity up to 37.56 3.13%. The results suggested ethnic heterogeneity with a significant gene cline with higher insertion allele frequency. Such population based data on various polymorphisms can ultimately be exploited in pharmacogenomics. PMID- 11927780 TI - Patient perceptions of quality of life and treatment in an outpatient congestive heart failure clinic. AB - In an effort to better understand patients' definitions of quality of life (QOL) and to determine which tools would be most appropriate for use in future studies, a descriptive study was done in a university-based congestive heart failure clinic. Participants were asked a series of five open-ended questions regarding their perceptions of QOL during recorded interviews. Most patients equated QOL with the ability to perform physical functions in the same way they did before developing heart failure. They grieved for their former abilities and expressed lower self-esteem due to loss of independence from physical limitations. The Short Form-36 and the Minnesota Living With Heart Failure Questionnaire addressed the QOL issues important to our patients. It is important for health care providers to consider the patient's perception of QOL when using quantitative tools for QOL measurement in clinical practice. PMID- 11927777 TI - Triplet repeat DNA structures and human genetic disease: dynamic mutations from dynamic DNA. AB - Fourteen genetic neurodegenerative diseases and three fragile sites have been associated with the expansion of (CTG)n (CAG)n, (CGG)n (CCG)n, or (GAA)n (TTC)n repeat tracts. Different models have been proposed for the expansion of triplet repeats, most of which presume the formation of alternative DNA structures in repeat tracts. One of the most likely structures, slipped strand DNA, may stably and reproducibly form within triplet repeat sequences. The propensity to form slipped strand DNA is proportional to the length and homogeneity of the repeat tract. The remarkable stability of slipped strand DNA may, in part, be due to loop-loop interactions facilitated by the sequence complementarity of the loops and the dynamic structure of three-way junctions formed at the loop-outs. PMID- 11927779 TI - Effective observation unit treatment of decompensated heart failure. AB - There is little information on the effectiveness of emergency department (ED) observation unit (OU) heart failure (HF) therapy. The authors' objective was to evaluate outcomes after implementation of an ED-OU treatment protocol for HF exacerbation. Unblinded assessment of the effectiveness of an HF protocol was performed, controlled by outcome for 9 months prior to implementation. This included diagnostic and therapeutic algorithms, cardiology consultation, close monitoring, patient education, and discharge planning. Adverse outcomes were defined as the 90-day rates of ED HF revisits, hospital HF readmissions, or death, as determined by chart review, computer database search, and phone follow up. One hundred fifty-four patients were enrolled; 50 entered before, and 104 after protocol implementation. Only six (12%) in the preprotocol and one (1%) of the postprotocol group were lost to follow-up. After an OU visit, postprotocol 90 day ED HF revisit rates declined 56% (0.90-0.51; p<0.0000) and the 90-day HF rehospitalization rate decreased 64% (0.77-0.50; p=0.007). The 90-day rates of death and OU HF readmission decreased from 4% to 1% (p=0.096) and 18% to 11% (p=0.099), respectively. An intensive outpatient ED OU HF management protocol safely decreases 90-day rates of emergency department visits and inpatient hospitalizations. PMID- 11927781 TI - Pharmacotherapy in congestive heart failure: diuretic resistance and strategies to overcome resistance in patients with congestive heart failure. AB - Congestive heart failure is a complex clinical hemodynamic disorder characterized by chronic and progressive pump failure and fluid accumulation. Although the overall impact of diuretic therapy on congestive heart failure mortality remains unknown, diuretics remain a vital component of symptomatic congestive heart failure management. Over time, sodium and water excretion are equalized before adequate fluid elimination occurs. This phenomenon is thought to occur in one out of three patients with congestive heart failure on diuretic therapy and is termed diuretic resistance. In congestive heart failure, both pharmacokinetic and pharmacodynamic alterations are thought to be responsible for diuretic resistance. Due to disease chronicity, symptomatic management is vital to improved quality of life and enhancing diuretic response is therefore pivotal. PMID- 11927776 TI - Molecular genetics of schizophrenia: past, present and future. AB - Schizophrenia is a severe neuropsychiatric disorder with a polygenic mode of inheritance which is also governed by non-genetic factors. Candidate genes identified on the basis of biochemical and pharmacological evidence are being tested for linkage and association studies. Neurotransmitters, especially dopamine and serotonin have been widely implicated in its etiology. Genome scan of all human chromosomes with closely spaced polymorphic markers is being used for linkage studies. The completion and availability of the first draft of Human Genome Sequence has provided a treasure-trove that can be utilized to gain insight into the so far inaccessible regions of the human genome. Significant technological advances for identification of single nucleo-tide polymorphisms (SNPs) and use of microarrays have further strengthened research methodologies for genetic analysis of complex traits. In this review, we summarize the evolution of schizophrenia genetics from the past to the present, current trends and future direction of research. PMID- 11927782 TI - Improving heart failure care: CMS' initiative. Improving heart failure care in outpatient practices. AB - This column is the eighth in a series reporting on the efforts of the Centers for Medicare & Medicaid Services ([CMS], formerly known as the Health Care Financing Administration), to improve care for Medicare beneficiaries with heart failure. Previous columns have focused on the hospital-based National Heart Failure project. An outpatient practice-based project, the Heart Failure Practice Improvement Effort (HF PIE), was described in the fourth and sixth columns. This column reports experience from the HF PIE project at the practice level in three states. PMID- 11927783 TI - Frontiers in congestive heart failure: heart failure in a cold climate. Seasonal variation in heart failure-related morbidity and mortality. PMID- 11927784 TI - Frontiers in congestive heart failure: initial experience with an implantable cardioverter-defibrillator incorporating cardiac resynchronization therapy. PMID- 11927785 TI - Difficult cases in heart failure: amiodarone lung injury: another heart failure mimic? AB - Careful thought must be given to the development of bystander pathology that could mimic worsening of heart failure. Recent trials with patients receiving amiodarone record a low rate of amiodarone pulmonary toxicity of 1.6%. Bronchoalveolar lavage in amiodarone toxicity demonstrates an absolute and relative lymphocytic alveolitis, suggesting hypersensitivity, but this finding is neither sensitive nor specific. Recently, KL-6, a mucin-like high molecular weight glycoprotein secreted by proliferating type II alveolar pneumocytes, has been identified as a potential marker of interstitial pneumonitis. A high index of suspicion combined with rapid exclusion of common confounding mimics can help in establishing the diagnosis of amiodarone lung toxicity. PMID- 11927787 TI - More on isolated systolic hypertension, diabetes, nephropathy and hypertension, and use of low-sodium diets in obese hypertensives. PMID- 11927786 TI - Nuggets, pearls, and vignettes of master heart failure clinicians. Part 4- treatment. PMID- 11927788 TI - Physician practice patterns in the treatment of isolated systolic hypertension in a primary care setting. AB - The authors evaluated the treatment of isolated systolic hypertension based on medical record review of charts between 1998 and 1999 in a multispecialty physician practice group. Two age-stratified random samples of ambulatory medical records were examined (393 patients aged > or =65 years and 251 patients aged 50 64 years). The samples corresponded to the practices of 35 primary care physicians who were surveyed about their hypertension care. Isolated systolic hypertension was defined as systolic blood pressure > or =140 mm Hg and diastolic blood pressure <90 mm Hg. Results showed that isolated systolic hypertension represented 76% and 45% of uncontrolled blood pressure in the older and middle aged samples, respectively. Isolated systolic hypertension was often undiagnosed and untreated. Physicians reported treatment thresholds and goals that were significantly less aggressive for their patients > or =65 years of age. Physician awareness and treatment of isolated systolic hypertension have not yet caught up with consensus guidelines, and older patients may be affected most by this gap. PMID- 11927789 TI - The effect of a losartan-based treatment regimen on isolated systolic hypertension. AB - This study was conducted to compare the antihypertensive efficacy and tolerability, over 12 weeks, of a losartan-based treatment regimen and placebo in patients with isolated systolic hypertension. Three hundred eight patients > or =35 years of age with isolated systolic hypertension, defined as trough sitting blood pressure between 140 and 200 mm Hg systolic and between 70 and 89 mm Hg diastolic, were randomized to losartan 50 mg (n=157) or placebo (n=151) once daily, with titration as necessary to achieve a goal trough sitting systolic blood pressure (SBP) <140 mm Hg. At baseline, mean trough sitting SBP was 140-159 mm Hg in 20.5% of patients, 160-179 mm Hg in 62.7%, and 180-200 mm Hg in 16.9%, and was similar in the two groups (losartan, 165.3 mm Hg; placebo, 166.1 mm Hg). At 12 weeks, mean trough sitting SBP decreased significantly (p<0.001) in both the losartan-based treatment group (by 19.2 mm Hg) and in the placebo group (by 7.6 mm Hg). The reduction in sitting SBP was significantly greater for losartan than placebo (-11.6 mm Hg; 95% confidence interval, -14.8 to -8.4). In patients with isolated systolic hypertension, a once-daily losartan-based treatment regimen significantly lowered SBP. The losartan-based regimen exhibited antihypertensive efficacy that was superior to that of placebo, with a similar tolerability profile. PMID- 11927790 TI - Hypertension in the elderly: a review of the importance of systolic blood pressure elevation. AB - The elderly, those 65 years of age and older, will account for 20% of the population by the year 2040. Hypertension affects more than one half of the elderly and its prevalence continues to increase with age. The presence of hypertension confers an increased risk of stroke, congestive heart failure, coronary heart disease, end-stage renal disease, and death. Although both diastolic and systolic blood pressure elevations are independently associated with increased cardiovascular risk in the younger individual, as vascular compliance becomes reduced at age 60, an increasing systolic blood pressure and lower diastolic blood pressure (or wider pulse pressure) increase cardiovascular risk in the elderly. Isolated systolic hypertension is the most common form of hypertension seen in the elderly. Lifestyle modification, including weight loss and salt restriction, reduces blood pressure, and may decrease the need for pharmacologic therapy. When the systolic blood pressure is 160 mm Hg or more and the diastolic blood pressure is <90 mm Hg, the initial use of a diuretic-based or calcium channel blocker-based regimen improves outcome. Alpha blocker therapy should not be used as initial monotherapy. Most elderly hypertensives will require two to three drugs to achieve the present blood pressure goal of <140 mm Hg. Caution should be exercised when lowering diastolic pressure to <55 mm Hg in those with isolated systolic hypertension. Although side effects of therapy are no more likely to occur in the elderly than in the younger individual with hypertension, blood pressure reduction should be accomplished gradually. PMID- 11927791 TI - Labetalol-induced hepatitis in a patient with chronic hepatitis B infection. AB - Drug-induced hepatotoxicity is an important cause of hepatocellular injury. Hepatic necrosis may range from asymptomatic elevations in transaminases to fulminant hepatic failure and death. A 50-year-old Asian man developed moderate elevations of hepatic transaminases after commencing labetalol therapy for a hypertensive cerebral hemorrhage. Extensive evaluation revealed evidence of chronic hepatitis B infection, of which the patient was previously not aware. Hepatic transaminases normalized after discontinuation of labetalol. PMID- 11927795 TI - ACE inhibitors and stroke: new considerations. PMID- 11927797 TI - Treating hypertension in the patient who has had a stroke. PMID- 11927796 TI - Diet wars: from Atkins to the Zone. Who is right? PMID- 11927798 TI - Transdermal clonidine skin reactions. AB - The clonidine transdermal therapeutic system is being used as a therapy for blood pressure treatment. Systemic side effects seem to be fewer than with oral clonidine. However, localized skin reactions occur frequently and the incidence increases with the dose and duration of use. Common signs include erythema, scaling, vesiculation, excoriation, and induration. Allergic contact dermatitis is less frequent but common. Hyperpigmentation and depigmentation also occur. Pretreatment with 0.5% hydrocortisone is associated with less skin irritation and higher blood levels. Although oral clonidine is no longer widely used in the management of hypertension, awareness of these reactions to the transdermal delivery of this agent is important. PMID- 11927799 TI - Diagnosis and treatment of supine hypertension in autonomic failure patients with orthostatic hypotension. AB - Orthostatic hypotension is seen in various medical conditions. It can be secondary to medications or volume depletion. It can also be due to autonomic neuropathy secondary to other diseases, such as diabetes mellitus, or to primary degenerative processes of the autonomic nervous system. Orthostatic hypotension dominates the clinical picture of patients suffering from autonomic failure. Paradoxically, about one half of these patients also suffer from supine hypertension, which induces pressure natriuresis, worsening orthostatic hypotension. It also complicates the treatment of orthostatic hypotension. Supine hypertension is mediated by an increase in peripheral vascular resistance. This is due to residual sympathetic tone in patients with multiple system atrophy (Shy Drager syndrome), but the cause is not known in patients with pure autonomic failure, who have increased vascular resistance despite very low levels or plasma norepinephrine and renin activity. The recent observation that patients with supine hypertension develop left ventricular hypertrophy suggests they should be treated. During the day, avoiding the supine position is often all that is required. Short-acting vasodilators (e.g., transdermal nitroglycerin) can be used during the night. PMID- 11927800 TI - Advicor (niacin extended-release and lovastatin tablets). PMID- 11927801 TI - Plate fixation of talar neck fractures: preliminary review of a new technique in twenty-three patients. AB - OBJECTIVE: To describe an unreported technique of fixation for talar neck fractures. DESIGN: Retrospective review and telephone interview of patients with fractures of the talar neck (OTA 72-A1.2, 72-A1.3, 72-C2). SETTING: Level-one trauma center with six trauma surgeons. PATIENTS/PARTICIPANTS: Twenty-three patients treated with minifragment plates over a four-year period. Follow-up intervals were an average of twenty months. INTERVENTION: Talar neck fractures were stabilized with one or two 2.0 or 2.4 plates, with additional 2.0, 2.7, or 3.5 lag screws when necessary. Most fractures were approached through medial and lateral incisions. MAIN OUTCOME MEASURES: Union rates, pain, and incidence of complications. Functional outcome will be best determined by a further review with longer follow-up. RESULTS: Mini fragment plates were placed on the side with the most comminution, and all wounds and fractures healed uneventfully. Four patients underwent hardware removal. Two patients developed a mild extension malunion, but there was no evidence of varus malunion, as measured on Canale views. CONCLUSIONS: Initial review suggests that plate fixation of comminuted talar neck fractures is a successful technique, with low rates of complications compared with those discussed in the literature. A further review in a few years will allow functional assessment studies. PMID- 11927802 TI - Treatment of complex acetabular fractures through a modified extended iliofemoral approach. AB - OBJECTIVES: To assess the rate of anatomic reconstructions as well as approach related morbidity and complications in the treatment of complex acetabular fractures through a modified extended iliofemoral approach. DESIGN: Prospective clinical study. SETTING: Level I trauma center, University Hospital. PATIENTS: Inclusion criteria were as follows: (a) associated acetabular fracture or transverse fracture with comminuted roof area stated as not sufficiently reconstructable through a single approach, and (b) age between sixteen and sixty five years. A total of forty-nine patients with fifty complex acetabular fractures could be included out of the series of ninety-six acetabular fractures treated operatively from August 1992 to February 1996. Open reduction and internal fixation of complex acetabular fractures through the modified extended iliofemoral approach were performed. RESULTS: In 80 percent of the fifty fractures the reduction was anatomic with a remaining displacement of less than or equal to one millimeter, in eight cases there was a persistent displacement of two millimeters, and two fractures had a poor result with a three-millimeter displacement. Complications included 8 percent loss of reduction, 13 percent heterotopic ossification grade 3, and 4 percent avascular femoral head necrosis. At the two-year follow-up there were 74 percent good or excellent radiographic and clinical results. Two patients had already been reoperated with total hip replacement, and the two patients with femoral head necrosis are currently scheduled for arthroplasty. CONCLUSIONS: The modified extended iliofemoral approach proved to be appropriate to achieve anatomic reduction in complex acetabular fractures. The high rate of approach-related morbidity has to be considered carefully and may lead to a decreased incidence of extended approaches. PMID- 11927803 TI - Second generation intramedullary nailing of subtrochanteric femur fractures: a biomechanical study of fracture site motion. AB - OBJECTIVES: To compare fracture site motion between different second-generation intramedullary nails used to fix subtrochanteric fractures of the proximal femur with and without femoral neck fractures. DESIGN: Nondestructive mechanical testing of four types of femoral intramedullary nails was undertaken to evaluate fracture site motion using a model that simulated single-leg and double-leg stance. METHODS: Three types of reconstruction nails (the Russell-Taylor Delta [Smith & Nephew, Memphis, TN], the Uniflex [Biomet, Warsaw, IN], Alta CFX [Howmedica-Osteonics, Rutherford, NJ]) and the Long Gamma nail (Howmedica Osteonics, Rutherford, NJ), each measuring 11 x 380 mm, were inserted in fiberglass composite femurs. Four fracture patterns were studied (transverse subtrochanteric, subtrochanteric with posteromedial wedge comminution, subtrochanteric with one-centimeter gap, and a one-centimeter gap with a subcapital neck fracture). Single-and double-leg stance loading was simulated using a servohydraulic load frame (MTS, Eden Prairie, MN). Two-way analysis of variance and post hoc t tests were used to determine any statistically significant differences between groups. RESULTS: In single-leg stance there were significant differences in coronal plane rotation, shear, and axial translation across the subtrochanteric fracture site between the different nail types and the different fracture patterns (p < 0.001). In double-leg stance there were significant differences in coronal plane rotation and femoral head vertical motion between the different nail types and the different fracture patterns (p < 0.001), and there were significant differences in shear and axial translation between the different fracture patterns (p < 0.001) but not the different nail types (p > 0.05). CONCLUSIONS: For simple, well-reduced fractures the choice of implant is not critical. As fracture severity increased (comminution, gap, and combined neck fracture), the choice of implant, particularly with reference to proximal nail dimensions and implant materials, was a significant factor in reducing fracture site motion. Therefore, our laboratory data suggest that when subtrochanteric fractures are unstable (e.g., comminution, segmental bone loss) and early weight bearing is desirable, the choice of implant is critical and should be restricted to implants that allow minimal fracture site motion (Long Gamma and Russell-Taylor). PMID- 11927805 TI - Late outcome of isolated tibial fractures treated by intramedullary nailing: the correlation between disease-specific and generic outcome measures. AB - OBJECTIVES: To measure the late morbidity of nailed isolated tibial fractures, using both a generic health score and disease-specific scores. To determine the correlation between the two types of outcome measure. DESIGN: Retrospective study, using a combination of case notes and radiographic review plus current clinical assessment. SETTING: Fracture Outcomes Research Unit in a U.K. teaching hospital. PATIENTS: Eighty-three patients with isolated fractures of the tibial diaphysis. Follow-up time was a minimum of three years from injury (mean fifty seven months). INTERVENTION: All patients were treated primarily by closed, reamed intramedullary nailing. OUTCOME MEASUREMENTS: Iowa Knee and Ankle Scores, visual analogue pain scores for fracture site and knee and ankle joints, and the Short Form 36 health status questionnaire. RESULTS: Sixty-four (77.1%) fractures united after the first procedure. Twenty-nine (34.9%) patients had pain around the knee at rest, fifty-nine (71.1%) had difficulty in kneeling, and thirteen (15.7%) were still experiencing some pain at their fracture site; 69% of patients had excellent results based on the Iowa scores as well as the SF-36 scores. Pain at the knee correlated with low Physical Component Summary and Mental Component Summary scores, and fracture site pain correlated with only low Physical Component Summary score. There was a significant correlation between the disease specific scores and the SF-36 scores, and only patients with an excellent Iowa grade had "normal" SF-36 scores. CONCLUSION: After tibial nailing, mild deficits registered by Iowa scores are associated with a significant disability and unhappiness as registered by the SF-36. According to the patient-oriented outcomes tool, 31% of late results are "unsatisfactory." PMID- 11927804 TI - Fat embolism and related effects during reamed and unreamed intramedullary nailing in a pig model. AB - OBJECTIVES: To determine whether reamed or unreamed femoral intramedullary nailing is more adverse to pulmonary function, the authors compared three populations of healthy pigs, analyzing the biochemical and hemodynamic effects related to fat embolism. Likewise, the authors histologically evaluated the presence of bone marrow fat embolism in lungs, heart, kidney, brain, and retina. DESIGN: Randomized, experimental model. SETTING: Laboratory. PARTICIPANTS: Twenty five male Duroc Jersey adult healthy pigs divided in three groups. INTERVENTION: Reamed and unreamed intramedullary nailing. OUTCOME MEASUREMENTS: Biochemical, hemodinamical, and histologic analysis. METHODS: In the first group of ten pigs, a reamed nail was inserted; in the second group of ten specimens, the authors placed an unreamed nail; and in the third group of five animals (control), only the surgical approach was made without opening the medullary cavity. RESULTS: The authors did not find statistically significant differences in pulmonary function between the reamed and unreamed group in the hemodynamic, biochemical, and histopathologic parameters evaluated. The histologic analysis of the lung tissue revealed a statistically significant difference between the nailed groups and the control (P < 0.04). CONCLUSIONS: In this animal model, the results indicate that pulmonary changes and fat embolization during intramedullary nailing occur to the same degree in reamed and in unreamed femurs. PMID- 11927806 TI - The effect of hemorrhagic shock in a caprine tibial fracture model. AB - OBJECTIVES: The purpose of this study was to assess effects of hemorrhagic shock on fracture healing in a large animal closed fracture model. DESIGN: Prospective randomized trial. SETTING: University Medical Center. STUDY SUBJECTS: Eight skeletally mature neutered male goats. INTERVENTION: Standardized bilateral closed midshaft tibia fractures were created in all the goats. The goats were randomized to a hemorrhage, shock and resuscitation group (shock group), or a control group. Hemorrhagic shock was induced in the four goats in the shock group. The shock state was maintained for thirty minutes. The remaining four goats were used as a control group. All fractures were stabilized with a standardized external fixator. OUTCOME MEASURES: Swan-Ganz catheters were used to record cardiac output (CO). Arterial blood was sampled to determine base deficit values for the goats after hemorrhage. Radiographs were obtained weekly. After four weeks the tibias were harvested and nondestructively tested in torsion. Immediately after mechanical testing, the tibias were processed for histologic analysis. Standard histomorphologic indices, including total bone volume (TV/BV%), osteoblast surfaces (Ob/BS%), and osteoid surface (OS/BS%), were measured. RESULTS: One goat in the shock group became agitated upon emergence from anesthesia and dislodged two of his external fixator pins. This animal was killed and was not included in the analysis of results. One goat that would have been entered into the control group was then switched to the shock group, leaving four goats in the shock group and three in the control group. One goat in the shock group developed a nonunion of his left tibia fracture. This nonunion occurred because of loosening of pin fixation in the distal tibia. The nonunited bone was not included in the analysis of results. The animal's right tibia healed uneventfully. Hemorrhage uniformly resulted in shock. In the shock group goats, average CO on induction of anesthesia was 3.7 l/min. Average CO dropped to 0.7 after hemorrhage and rose to 6.6 after fluid resuscitation. The average base deficit for the shock group animals was -9.9. The control group animals had minor drops in average CO, from 4.5 on induction of anesthesia to 4.3 on emergence from anesthesia. Radiographic analysis showed no apparent differences in healing between groups when comparing callus area and density. With the exception of the nonunion, all tibiae were considered to be healing normally radiographically. Biomechanical testing showed no statistical difference between the shock and control groups in maximum torque (p = 0.95), stiffness (p = 0.64), and energy absorbed at failure (p = 0.91). Histomorphologic results indicated there were no differences between groups. Shock did not appear to influence bone formation rate or callus remodeling compared with controls (p = 0.6). No evidence of osteocyte necrosis was observed. CONCLUSIONS: In this study no significant difference was found between the shock and control groups in any of the parameters measured. Transient hemorrhagic shock does not appear to adversely affect closed tibia fracture healing in a goat model. PMID- 11927807 TI - Improvement of the bone-screw interface strength with hydroxyapatite-coated and titanium-coated AO/ASIF cortical screws. AB - OBJECTIVE: To evaluate whether coating AO/ASIF screws with osteoconductive materials can improve bone-screw fixation. DESIGN: Ninety-six AO/ASIF 4.5 millimeter cortical screws were divided into four paired groups and implanted in the femurs and tibiae of six sheep: Group A = standard stainless steel screws; Group B = stainless steel screws coated with highly crystalline hydroxyapatite; Group C = stainless steel screws coated with low crystalline hydroxyapatite; Group D = titanium screws coated with titanium. The screws were implanted according to the standard AO technique to an insertion torque of 2,000 Newton millimeters. Sheep were killed at one, three, and twelve months after surgery. MAIN OUTCOME MEASURES: Extraction torque was measured on six screws from each group selected at random at time of each euthanization. Morphologic analysis of the bone-screw interface was performed on the remaining screws. RESULTS: At each euthanization the extraction torque of Group A was lower than that of the other groups (p < 0.0001). At three and twelve months the extraction torque of Group B was higher than that of Group D (p = 0.002). Morphologic results showed extensive bone-screw gap in Group A. Optimal osteointegration was observed in Groups B and C. Osteointegration of Group D was higher than that of Group A and lower than that of Groups B and C. CONCLUSIONS: It was demonstrated that AO/ASIF screws coated with osteoconductive materials achieve optimal fixation strength, even in the early phase. This fixation strength was significantly higher than that of the standard screws. PMID- 11927808 TI - Endoscopic access to the extremities: the principle of fascial clefts. AB - Technology for endoscopic surgery has developed rapidly during the last decade. Applications of endoscopic techniques to orthopaedic surgery have been made possible by the use of balloon dissectors. Balloon dissectors create an optical cavity by separating fascial layers of a constant anatomic plane called the fascial cleft. The optical cavity can be maintained with either carbon dioxide (CO2) insufflation or manual retractors. The authors of the present study have developed a safe, reliable technique using a balloon dissector to create such optical cavities in the extremities, pelvis, and acetabulum to facilitate minimally invasive surgery in these areas. The authors' clinical work and fresh cadaver dissection confirms that the fascial cleft is a universal anatomic constant. It can be accessed quickly to facilitate endoscopic procedures, such as bone grafting for delayed unions, tissue expansion for reconstructive surgery, sural nerve harvesting for nerve cable grafting, and microvascular tissue transfer harvesting and flap prefabrication for extremity reconstruction. Twenty five cases, each with an average follow-up of 34 months, are presented. Indications, results, and complications of balloon-assisted endoscopic surgery are described. PMID- 11927809 TI - Internal fixation of a patella fracture using an absorbable suture. AB - This case report describes the use of an absorbable suture for the fixation of a patella fracture in a child. In this case the fracture was held satisfactorily by the suture until union. This technique means that there is no need for a second operation to remove the fixation material. PMID- 11927810 TI - Traumatic avulsion of the triceps brachii. AB - Traumatic avulsion of the triceps brachii tendon is a rare injury. Clinical diagnosis shows that elbow extension against gravity is not possible. Radiologically, the lateral elbow may show the presence of flecks of avulsed osseous material from the olecranon ("flake sign"), which is almost pathognomonic of this lesion. The authors of the present study present a case of avulsion fracture treated with the attachment of bone suture anchors. PMID- 11927811 TI - Open iliac wing fracture caused by penetrating injury from a bicycle handlebar. AB - Bicycle accidents have been documented as one of the most common mechanisms of accidents in children. Several reports describe intra-abdominal injury secondary to bicycle handlebars. Reported injuries include liver and spleen trauma, bowel perforation, and pancreatitis. However, there are few reports of penetrating handlebar injuries. We report a case of a penetrating bicycle handlebar producing an open iliac wing fracture. A number of reports have stressed the dichotomy between the benign external appearance and the potential severity of the intra abdominal injury after a bicycle handlebar injury. Thus, a high index of suspicion must be maintained when evaluating a child after such an injury. All patients with a significant mechanism should have a thorough and complete evaluation, including radiographs. As with all traumatic injuries, a multidisciplinary approach is often required to efficiently manage these injuries. Although uncommon, open pelvic injuries in children do occur. The principles of management are the same in children as they are in adults. All children with open pelvic fractures require emergent operative debridement and, if indicated, stabilization. Special attention should be given to ensure that the gastrointestinal and genitourinary tracts do not communicate with the fracture. With attention to these fundamental principles, severe pelvic injuries in children can be managed with few long-term sequelae. PMID- 11927812 TI - Subtrochanteric femur fracture. PMID- 11927816 TI - Lumbar multilevel degenerative spondylolisthesis: radiological evaluation and factors related to anterolisthesis and retrolisthesis. AB - Radiographs of 3,259 outpatients with low back disorders were examined for age, gender, level, direction, degree of slip, lumbar lordosis, pedicle-facet (P-F) angle, facet shape, and disc height. Degenerative lumbar spondylolisthesis was found in 284 (8.7%) of the patients, of which 83 were excluded. Single-level spondylolisthesis was present in 132 of the 201 patients studied, including 93 cases of anterolisthesis and 39 of retrolisthesis, the former being predominant at L4 and in women and the latter at L2 and equal between the genders. Multilevel spondylolisthesis in 69 patients included 65 (94%) of two-segment slip, 21 anterior, 25 posterior, and 19 combined, and 4 cases of three-segment retrolisthesis. Factors related to anterolisthesis were increased P-F angle and W shaped facet joint; statistically, however, no factors were found statistically related to retrolisthesis. Multilevel anterolisthesis was considered to occur from factors similar to those previously reported for single-level anterolisthesis, and the pathomechanism of retrolisthesis is different from that of anterolisthesis. PMID- 11927817 TI - Evaluation of the fixation and strength of a "rescue" revision pedicle screw. AB - One potential complication of pedicle screw instrumentation is long-term nerve root irritation, most commonly resulting from medial or inferior screw malpositioning. This study evaluated the pullout strength and fatigue strength properties of a novel "rescue" revision screw designed to reduce the sequelae of medial screw malpositioning by eliminating threads along the section of the screw corresponding to the pedicular region. The results of this study revealed that a rescue screw with threads eliminated from one half of its circumference provided comparable pullout strength to fully threaded pedicle screws in cadaveric specimens. In addition, mechanical testing of the rescue screw did not show a decrease in fatigue strength characteristics compared with controls. These results suggest that the rescue screw may offer a way to decrease nerve root irritation caused by canal impingement without jeopardizing the overall strength of the spinal construct. PMID- 11927818 TI - Surgical management of lumbar degenerative spinal stenosis with spondylolisthesis via posterior reduction with minimal laminectomy. AB - Degenerative lumbar spondylolisthesis with spinal stenosis is commonly treated with laminectomy. Recent reports have consistently supported the incremental clinical benefit of associated in situ arthrodesis with or without instrumentation. Resection of the lamina may result in intraoperative dural tear or epidural scar formation. Fifty-six consecutive patients with back pain, neuroclaudication, or both, in addition to degenerative spondylolisthesis with spinal stenosis, underwent a surgical procedure that incorporated fusion after reduction of the spondylolisthesis deformity with preservation of the lamina and the balance of the posterior elements. Clinical records were reviewed and patients interviewed at a mean of 33 months after surgery. Oswestry Disability Index scores were obtained independently at baseline and at a late review. Late imaging was available a mean of 28 months after operation. Clinical and imaging analyses and Oswestry scoring confirmed results comparable to the published outcomes of in situ fusion after formal laminectomy. Resection of the lamina may not be necessary in the treatment of degenerative lumbar spinal stenosis with spondylolisthesis. PMID- 11927819 TI - Osteomyelitis of the cervical spine: a potentially dramatic disease. AB - Osteomyelitis of the cervical spine is a rare disease, representing only 3% to 6% of all cases of vertebral osteomyelitis. In contrast with other locations of spinal infections, osteomyelitis of the cervical spine can be a much more dramatic and rapidly deteriorating process, leading to early neurologic deficit. Thus, the disease must be diagnosed quickly and appropriate therapy initiated as soon as possible. The clinical course, therapy, and outcome of 15 patients treated for osteomyelitis of the cervical spine are presented. Nine of 15 patients presented with a neurologic deficit at the time of diagnosis. Surgical treatment consisted of radical debridement of the infected bone and either immediate bone grafting and stabilization as a one-step procedure or interval antibiotic treatment before bone grafting and surgical stabilization as a second procedure. A favorable outcome was achieved by early and aggressive surgical intervention, including complete resolution of neurologic deficits in more than 50% of the patients and complete bony fusion in all but one patient. The authors prefer additional posterior rather than anterior stabilization alone to perform fusion over a shorter distance involving only the infected segments. PMID- 11927820 TI - A histologic study of fractured human vertebral bodies. AB - Twenty-seven fractured human vertebral bodies and 24 unfractured human vertebrae from adjacent levels were studied postmortem using histologic and high-resolution radiographic techniques. The findings were compared with those in the vertebral bodies of individuals without spinal fracture. Forty-six human thoracolumbar spines were obtained from individuals at autopsy. Standard radiographs were made of all specimens. Twelve of the 46 individuals had a total of 27 fractured vertebral bodies by plain radiographic criteria. Attention was focused on these fractured vertebrae as well as on 24 unfractured vertebral bodies that were harvested from a level immediately adjacent to the fractured vertebral bodies. Twelve vertebral bodies from four individuals with no evidence of fracture or inflammatory spondyloarthropathy were also studied for comparison. The vertebral bodies were graded by their mineral density as measured by dual-energy x-ray absorptiometry and sectioned into 3-mm sagittal cuts. High-resolution contact radiographs were prepared for each section prior to decalcification and tissue sectioning on a large format microtome. Mid-and parasagittal tissue sections of each vertebra were prepared for standard hematoxylin and eosin stains. A total of 126 sections were studied. The histologic characteristics of the fractured vertebrae (n = 27) were compared with those of adjacent unfractured levels (n = 24) and with vertebrae from individuals without fracture (n = 12). Vertebral bodies with fractures secondary to osteoporosis were consistently characterized histologically by focal areas of endochondral new bone formation adjacent to avascular necrotic bone and unreactive marrow. Such ongoing new bone and new vessel formation adjacent to nonhealing areas were also documented in radiographically unfractured vertebral bodies from individuals with osteoporotic fractures at adjacent levels. No areas of endochondral new bone formation or areas of focal necrosis were found in vertebral bodies from individuals without radiographic evidence of osteoporosis. A vascular necrosis of the vertebral body is a common histologic finding in individuals with osteoporosis. Indeed, our histologic observations suggest subclinical fractures and microfractures of the vertebral body may be the underlying pathologic process leading to avascular necrosis in individuals with osteoporosis. Microtrabecular fractures and endplate fractures were commonly seen in osteoporotic vertebral bodies, often in vertebrae that appeared to be uninvolved on specimen radiographs. PMID- 11927821 TI - Surgical correction of the late consequences of posttraumatic spinal disorders. AB - This retrospective study was performed to evaluate the results of the treatment of late posttraumatic spinal disorders. Five patients with thoracolumbar kyphosis, two with paralytic kyphoscoliosis and two with posttraumatic instability with pain but without neurologic symptoms and apparent deformity, were treated surgically. Various surgical procedures were performed depending on the pathologic conditions. Corrections achieved in the sagittal and frontal planes were analyzed on radiographs after a mean follow-up of 4.4 years. In cases of angular kyphosis, the deformity measured 48.8 degrees on average before operation and 9.3 degrees afterward. Preoperative neurologic impairments were improved in all cases. Patients with paralytic scoliosis benefited from an average surgical correction of 68%. The restoration of truncal balance helped the patients to regain a normal sitting posture. Pain was reduced in all patients after surgery, as it was in patients with low back pain caused by posttraumatic instability without deformity. The results of the current study emphasize the importance of adequate management of spinal injuries to prevent late spinal sequelae. PMID- 11927822 TI - Bilateral pneumothoraces, pneumomediastinum, pneumoperitoneum, pneumoretroperitoneum, and subcutaneous emphysema following intubation with a double-lumen endotracheal tube for thoracoscopic anterior spinal release and fusion in a patient with idiopathic scoliosis. AB - Recently, thoracoscopic approaches to the spine have taken on greater clinical applications in the treatment of spinal deformity with generally good results. However, the steep learning curve must be ascended by the surgeon and may lead to complications early in one's experience. There also exists a learning curve for the anesthesiologist to become adept at obtaining single lung ventilation and managing this throughout the operative procedure. We report a case of an 11-year old patient with severe scoliosis who developed air in both chest cavities, mediastinum, peritoneum, retroperitoneum, and subcutaneous tissue after intubation with a double-lumen endotracheal tube. The patient remained hemodynamically stable throughout this period, and bilateral chest tubes were placed. The patient remained on the ventilator for 24 hours and was extubated without sequelae. Complications from a thoracoscopic approach to the spine for deformity are most often attributed to the learning curve of the surgeon; however, the entire operative team becomes exposed to the challenges of performing this procedure. This report documents a life-threatening complication of air throughout the chest, mediastinum, abdomen, and subcutaneous tissues in a patient with severe scoliosis. PMID- 11927823 TI - Flexion-distraction injuries in the thoracolumbar spine: an in vitro study of the relation between flexion angle and the motion axis of fracture. AB - A new concept, the motion axis of fracture (MAF), which is defined as the transitional point from anterior compressive to posterior splitting failure on a lateral radiograph, has provided a true understanding of the mechanisms of flexion-distraction injuries in clinical cases. This study was designed to produce in vitro injuries that have MAFs and to clarify the relation between the flexion angle and the MAF location. Adolescent porcine thoracolumbar spines were exposed to a vertical compressive load to failure at three different flexion angles and then examined radiographically. The MAF location was recorded as the distance from the anterior border to the MAF expressed as a percentage of the anteroposterior diameter of the vertebral body. All specimens showed similar injuries, with MAFs consisting of anterior compression fractures in the vertebral bodies and posterior disruptions. A significant negative correlation emerged between the flexion angle and the MAF location (r = -0.890; p < 0.0001). These results suggest that even a vertical compressive load contributes to the production of a flexion-distraction injury with an MAF in the thoracolumbar spine. They also indicate that the flexion angle of the spine at which the vertical compressive load is applied is an important factor in determining the MAF location; that is, the larger the flexion angle, the more anterior the MAF. PMID- 11927824 TI - Thoracic vertebral body exostosis as a cause of myelopathy in a patient with hereditary multiple exostoses. AB - The posterior thoracic vertebral body appears to be a novel origin for an exostosis causing myelopathy. A patient with hereditary multiple exostoses and myelopathy caused by an exostosis originating from the posterior aspect of the T5 vertebral body was treated with a staged anterior decompression/corpectomy and posterior spinal fusion. The patient had near-complete resolution of his myelopathy immediately after undergoing removal of the exostosis through a right sided lateral thoracotomy approach. This was a unique origin for an exostosis causing spinal cord compression in a patient with hereditary multiple exostoses. The delivery of the exostosis was performed en bloc during the anterior decompression and corpectomy portion of the surgery. This resulted in the expected favorable outcome. PMID- 11927825 TI - Better treatment of vertebral osteomyelitis using posterior stabilization and titanium mesh cages. AB - There is no scientific consensus on the role of posterior instrumentation in vertebral osteomyelitis. No study has examined the use of titanium cages to reconstruct the anterior column of the spine with vertebral osteomyelitis. Here the authors evaluated the efficacy of using titanium mesh cages anteriorly and posterior instrumentation after anterior debridement in the surgical treatment of vertebral osteomyelitis. In one center, 21 consecutive patients had surgery for vertebral osteomyelitis. The mean follow-up time was 67 months (range, 24 to 120 months). Ten patients received supplemental posterior instrumentation. Five patients had reconstruction of the anterior column with titanium cages. Greater improvement in sagittal alignment was noted for patients with cages implanted (p = 0.0009) and for those with posterior instrumentation (p = 0.005). Patients who received cages had greater (p = 0.0006) correction of their coronal alignment than did those patients without cages. A trend toward fewer postoperative complications emerged for patients who had posterior stabilization or titanium cages. These results support the use of posterior stabilization and titanium cages in the surgical treatment of vertebral osteomyelitis. PMID- 11927827 TI - An in vivo MRI study of the changes in volume (and fluid content) of the lumbar intervertebral disc after overnight bed rest and during an 8-hour walking protocol. AB - Magnetic resonance imaging (MRI) was used to measure the changes in the volume (and fluid content) of the lumbar intervertebral discs (L1-L2, L2-L3, L3-L4, L4 L5) in five normal subjects. For each subject, MRI scans were taken at the end of a normal day and again on the following morning (after a night's bed rest). Ten further scans were taken during an 8-h protocol consisting of alternate periods of walking (40 min) and scanning (10 min). On average, 1) disc volume increased by 10.6% during overnight bed rest, which corresponds to a gain of about 0.9 cm(3) of fluid; 2) the rate of disc volume decrease during the 8-h walking protocol was 0.96 x 10(-3) cm(3)/min; and 3) after 8 h (using our walking/scanning protocol), the disc volume did not decrease to the volume measured at the end of the previous day. PMID- 11927828 TI - Synthesis of a Kunitz-type serine proteinase inhibitory protein that shares homology with bovine pancreatic trypsin inhibitor by ovine intervertebral disc cells in serum-free alginate bead culture. AB - The objective of this study was to determine whether disc cells could be cultured under serum-free conditions and whether they synthesized bovine pancreatic trypsin inhibitor (BPTI)-like serine proteinase inhibitory proteins (SPIs) previously demonstrated for ovine chondrocytes. Intervertebral discs from 1- to 2 year-old merino wether sheep were dissected into the annulus fibrosus and nucleus pulposus, and the cells isolated by sequential enzymatic digestion. The cells were grown encapsulated in calcium alginate microspheres under serum-free conditions for 10 days. They remained more than 92% viable as assessed using the vital fluorescent dyes chloromethyl fluorescein diacetate and ethidium homodimer 1 to delineate live/dead cells, respectively. Western and affinity blotting identified a 12-16-kDa media SPI and an additional 34-36-kDa BPTI-like species in solubilized bead samples. This study has demonstrated ovine disc cells synthesized BPTI-like SPIs in serum-free alginate bead culture similar to chondrocyte SPIs; however, the 58-kDa precursor SPI form was not detected suggesting differences in the endogenous processing of these SPIs. PMID- 11927829 TI - Postoperative Clostridium perfringens lumbar discitis with septicemia: report of a case with survival. AB - This case report describes the successful treatment of a case of Clostridium perfringens postoperative discitis and suggests a management protocol for this condition. The medical literature to date reports only one similar case, treated without success. The patient has been well but the affected space later collapsed with mechanical pain and has required fusion. C. perfringens discitis can be treated successfully. PMID- 11927830 TI - Role of surface roughness of titanium versus hydroxyapatite on human bone marrow cells response. AB - This study was designed to assess human bone marrow cell response and particularly cell adhesion, proliferation, and differentiation, when cultured in vitro on titanium alloy and hydroxyapatite with different values of surface roughness. A further aim was to compare the cell response on the two materials, currently used in spinal surgery. Cell adhesion was determined after 0.5, 2, 4, and 18 hours of incubation; proliferation after 8, 11, 14, and 16 days of culture; and differentiation was evaluated with the expression of alkaline phosphatase activity after 8 and 16 days of culture. This study showed that bone marrow cells grew faster on titanium alloy than on hydroxyapatite, although fewer cells attached to titanium, compared to those attached to hydroxyapatite. No statistically significant difference was observed as the expression of alkaline phosphatase activity on hydroxyapatite and titanium alloy of the same roughness. Cell adhesion, proliferation, and differentiation are dependent on surface roughness of the biomaterial, and all three increased as the roughness of titanium alloy increased. Conclusively, surface roughness of titanium and hydroxyapatite significantly influences bone marrow cell response, and therefore these biomaterials should be used with rough outer surface, if a significant cell response on them is desired. These advantages of titanium and hydroxyapatite theoretically seem to be of particular importance in the following situations: long fusions, lumbosacral fusion, revision surgery with poor bone bank, neuropathic scoliosis associated with few bone graft reserves, and adult patients with severe osteoporosis. PMID- 11927832 TI - Differing effects of methylenetetrahydrofolate reductase single nucleotide polymorphisms on methotrexate efficacy and toxicity in rheumatoid arthritis. PMID- 11927833 TI - Polymorphisms in the methylenetetrahydrofolate reductase gene were associated with both the efficacy and the toxicity of methotrexate used for the treatment of rheumatoid arthritis, as evidenced by single locus and haplotype analyses. AB - 5,10-Methylenetetrahydrofolate reductase (MTHFR), a key enzyme involved in folate metabolism, has two common polymorphisms that affect enzyme activity. The objective of this study was to examine whether there was a correlation between the genotype or haplotype of the MTHFR gene and the efficacy or toxicity of methotrexate (MTX) in the treatment of rheumatoid arthritis. MTX-treated rheumatoid arthritis patients (n = 106) were selected from outpatient clinics and used for a retrospective study to examine the correlation between genotypes or haplotypes concerning polymorphisms of the MTHFR gene, and the efficacy or toxicity of MTX. Estimation of the haplotype frequencies was performed by maximum likelihood estimation based on expectation maximization algorithm. Single locus analysis examining each locus separately showed that patients with 1298C were receiving significantly lower doses of MTX compared to patients without [P < 0.05, relative risk (RR) = 2.18, 95% confidence interval (CI) 1.17-4.06], while a higher rate of overall MTX toxicity was observed in patients with 677T than those without (P < 0.05, RR = 1.25, 95% CI 1.05-1.49). An estimation of haplotype frequencies showed that there was no 677T-1298C haplotype in the population. Posterior distribution of the diplotype configuration for each individual was concentrated on a single configuration. Patients with the 677C-1298C haplotype were receiving lower doses of MTX than those without (P < 0.05, RR = 2.14, 95% CI 1.13-4.07), while subjects with 677T-1298A had a higher frequency of side-effects from MTX (P < 0.05, RR = 1.42, 95% CI 1.11-1.82). Both single locus and haplotype analyses suggest that polymorphisms within the MTHFR gene are associated with both the efficacy and toxicity of MTX in rheumatoid arthritis patients. Pharmacokinetic studies are necessary to prove the association. PMID- 11927834 TI - Molecular analysis of thiopurine S-methyltransferase alleles in South-east Asian populations. AB - Thiopurine S-methyltransferase (TPMT) catalyses the S-methylation of thiopurine drugs. In Caucasians, four variant TPMT alleles have been detected in over 80% of individuals with low or intermediate TPMT activity. The wild-type allele is designated as TPMT*1 and the mutant alleles are designated TPMT*2 through TPMT*8. The frequency of these alleles in different ethnic groups has not been well defined. In this study, one hundred individuals, from each of the Indonesian, Thai and Philippine populations, together with 249 Taiwanese, were analysed by polymerase chain reaction-restriction fragment length polymorphism and direct sequencing methods. The results showed that the allelic frequencies of TPMT*3C were 0.6% for Taiwanese and 1% for Filipino, Thai and Indonesian populations, respectively. One Filipino with a Caucasian parent was found to be heterozygous for the TPMT*2 allele. This study provides the first analysis of the allele frequency distribution of all known TPMT mutations in South-east Asian populations. PMID- 11927835 TI - Variability in response to nicotine in the LSxSS RI strains: potential role of polymorphisms in alpha4 and alpha6 nicotinic receptor genes. AB - Several studies have shown that genetic factors influence the effects of nicotine on respiration, acoustic startle, Y-maze crosses and rears, heart rate and body temperature in the mouse. Recently, we identified restriction fragment length polymorphisms (RFLPs) associated with the alpha4 (Chrna4) and alpha6 (Chrna6) nicotinic cholinergic receptor genes in the recombinant inbred (RI) strains derived from the Long-Sleep (LS) and Short-Sleep (SS) mouse lines. The alpha4 polymorphism has been identified as a point-mutation at position 529 (threonine to alanine) and the alpha6 polymorphism has not yet been identified. The studies described here evaluated the potential role of these polymorphisms in regulating sensitivity to nicotine by constructing dose-response curves for the effects of nicotine on six responses in the LSxSS RI strains. The results obtained suggest that both of the polymorphisms may play a role in regulating variability in sensitivity to nicotine. Those RI strains carrying the LS-like alpha4 RFLP were significantly more sensitive to the effects of nicotine on Y-maze crosses and rears, temperature and respiration and were less sensitive to the effects of nicotine on acoustic startle than those strains carrying the SS-like alpha4 RFLP. Those RI strains carrying the LS-like alpha6 RFLP were more sensitive to the effects of nicotine on respiration and acoustic startle, and less sensitive to the effects of nicotine on Y-maze crosses than those strains carrying the SS-like alpha6 RFLP. These results suggest that genetically determined differences in sensitivity to nicotine may be explained, in part, by variability associated with at least two of the neuronal nicotinic receptor genes, alpha4 and alpha6. PMID- 11927836 TI - Identification and ethnic distribution of major haplotypes in the gene GNB3 encoding the G-protein beta3 subunit. AB - The 825T-allele of the gene GNB3 encoding the G protein beta3 subunit is associated with hypertension and obesity, and identifies individuals highly responsive to diuretic therapy. Gbeta3s, a Gbeta3 protein variant generated by alternative splicing in carriers of the 825T-allele, is linked to increased signal transduction and is a potential cause for the observed pathophysiology. Here, we searched the entire GNB3 gene for additional polymorphisms and analysed their prevalence in Caucasian, black African and Asian populations. We detected six novel single nucleotide polymorphisms which were termed according to their location as G76A, G1906T, G2906A, A3882C, G5177A, and G5249A. Furthermore, we found a CACA-insertion-deletion polymorphism at position 6496. Genotyping and association studies resulted in the definition of two major GNB3 haplotypes, termed 'C-haplotype' (alleles 825C, 3882A, 5249G, 6496CACA-) and 'T-haplotype' (alleles 825T, 3882C, 5249A, 6496CACA+). Molecular modelling studies revealed that the pre-mRNA structures of both haplotypes exhibit marked differences which may account for the alternative splicing predominantly observed with the T haplotype. The prevalence of these haplotypes in major ethnic populations differs considerably. Furthermore, we detected additional frequent GNB3 polymorphisms. These variants were restricted to one or two major ethnic populations. Our results will aid future studies on population-specific effects of the GNB3 variants on risk and course of frequent diseases, including hypertension, obesity, stroke and myocardial infarction. Furthermore, they will contribute to the understanding of GNB3-related population-specific pharmacogenetic differences in the response to major drugs, as already shown for diuretics and antidepressants. PMID- 11927838 TI - N-Acetyltransferase genotypes as modifiers of diisocyanate exposure-associated asthma risk. AB - We observed previously that polymorphisms in glutathione S-transferase (GST) genes modified allergic responses to diisocyanate exposure. Here, we extended the study to examine the possible role of N-acetyltransferase (NAT) genotypes in the development of diisocyanate-induced ill effects, both separately and in combination with the previously examined GSTM1, GSTM3, GSTP1 and GSTT1 genotypes. The study population comprised 182 diisocyanate-exposed workers, 109 of whom were diagnosed with diisocyanate-induced asthma and 73 of whom had no symptoms of asthma. The diisocyanates to which the workers had been exposed to were diphenylmethane diisocyanate (MDI), hexamethylene diisocyanate (HDI) and toluene diisocyanate (TDI). The NAT2 genotype did not have any significant effect on the risk of developing asthma, but the putative slow acetylator NAT1 genotypes posed a 2.54-fold risk of diisocyanate-induced asthma (95% confidence interval [CI] 1.32 to 4.91). The effect of the NAT1 genotype was especially marked for workers exposed to TDI, among whom the NAT1 slow acetylator genotypes posed a 7.77-fold risk of asthma (95% CI 1.18 to 51.6). Statistically significant increases in asthma risk were also observed among the whole study population for the concurrent presence of the GSTM1 null genotype and either NAT1 (odds ratio [OR] 4.53, 95% CI 1.76 to 11.6) or NAT2 (OR 3.12, 95% CI 1.11 to 8.78) slow acetylator genotypes, and of NAT1 and NAT2 slow acetylator genotypes (OR 4.20, 95% CI 1.51 to 11.6). The results suggest for the first time that in addition to GSTs, the NATs play an important role in inception of asthmatic reactions related to occupational exposure to diisocyanates. PMID- 11927839 TI - Antipsychotic-induced extrapyramidal syndromes and cytochrome P450 2D6 genotype: a case-control study. AB - To study the association between polymorphism of the cytochrome P450 2D6 gene (CYP2D6) and the risk of antipsychotic-induced extrapyramidal syndromes, as measured by the use of antiparkinsonian medication. Data for this case-control study were obtained from a psychiatric hospital where newly admitted patients are routinely screened for several CYP2D6 mutant alleles. Cases were patients prescribed antiparkinsonian medication during oral antipsychotic drug treatment in the period September 1994 to August 2000. They were divided into those using an antipsychotic drug the metabolic elimination of which depends on the activity of the CYP2D6 enzyme ('CYP2D6-dependent') and those using other antipsychotic drugs. We formed a control group of antipsychotic drug users for both case groups using a matching ratio of 3 : 1 (controls : cases). Control patients were matched on whether or not their prescribed antipsychotic drug was CYP2D6-dependent. Odds ratios for patients who were slow metabolizers versus patients who were extensive metabolizers were calculated using conditional logistic regression and were adjusted for age, gender, dose and other potential confounding factors. We identified 77 case patients who were prescribed a CYP2D6-dependent antipsychotic drug and 54 case patients who were prescribed non CYP2D6-dependent antipsychotic drugs. Among the case- and control-patients using a CYP2D6-dependent antipsychotic drug, the poor metabolizers were more than four times more likely to start with antiparkinsonian medication than the extensive metabolizers (odds ratio 4.44; 95% confidence interval 1.11-17.68). An increased risk was not observed for patients using non CYP2D6-dependent antipsychotic drugs (odds ratio 1.20; 95% confidence interval 0.21-6.79). Genetically impaired CYP2D6 activity can increase the risk of antipsychotic-induced extrapyrimidal syndromes. Poor metabolizers should have their antipsychotic drug dosage reduced when the metabolism of the prescribed drug depends on CYP2D6 activity or should receive an antipsychotic drug that is not CYP2D6-dependent. PMID- 11927837 TI - Analysis of the CYP2C19 polymorphism in a North-eastern Thai population. AB - CYP2C19 is a polymorphically expressed cytochrome P450 responsible for the metabolism of several clinically used drugs, including some barbiturates, diazepam, proguanil, propranolol and several proton pump inhibitors. Genetic polymorphism of this enzyme shows marked interracial differences, with the poor metabolizer (PM) phenotype representing 2-5% of Caucasian and 11-23% of Oriental populations. In the present study, CYP2C19 phenotype and genotype were investigated in 107 North-eastern Thai subjects using the omeprazole hydroxylation index (HI) and polymerase chain reaction-restriction fragment length polymorphism technique, respectively. It was found that the distribution of HI in these subjects was bimodal. Seven subjects [6.54%, 95% confidence (CI) 1.86-11.22%] were identified as PM, with an HI > 7. Analysis of CYP2C19 genotypes in these 107 Thai subjects revealed that the allele frequencies for CYP2C19*1, CYP2C19*2 and CYP2C19*3 were 0.71 (95% CI 0.65-0.77), 0.27 (95% CI 0.21-0.33) and 0.02 (95% CI 0.01-0.05), respectively. The PM phenotype and the frequencies of CYP2C19 defective alleles in Thais, particularly CYP2C19*3, were lower than those observed in other Oriental populations. It is noteworthy that there was a case of nonaccordance between phenotype and genotype in one of the PMs. Whether this PM represents a novel defective allele requires further investigation. PMID- 11927840 TI - Variation in coumarin 7-hydroxylase activity associated with genetic polymorphism of cytochrome P450 2A6 and the body status of iron stores in adult Thai males and females. AB - The relationships between catalytic activity of cytochrome P450 2A6 (CYP2A6), polymorphism of CYP2A6 gene, gender and levels of body iron stores were analysed in a sample group of 202 apparently healthy Thais, aged 19-47 years. Eleven individuals were found to have high activity of CYP2A6, judged by the relatively large amounts (11.2-14.6 mg) of 7-hydroyxcoumarin (7-OHC) excreted 3 h following administration of 15 mg of coumarin. Ten individuals, however, did not excrete any 7-OHC. Of these 10, four were found to have no CYP2A6 gene (whole gene deletion; CYP2A6*4 allele). The frequency of the CYP2A6 alleles; *1A, *1B and *4 in the whole sample group was 52, 40 and 8% while the frequency of the CYP2A6 gene types; *1A/*1A, *1A/*1B, *1B/*1B, *1A/*4, *1B/*4, *4/*4 was 29, 41, 16, 7, 5 and 2%. Subjects having CYP2A6*1A/*1B gene-type group were found to have higher rates of coumarin 7-hydroxylation compared with those of the CYP2A6*1B/*1B and CYP2A6*1A/*4 gene types. The inter-individual variability in CYP2A6 catalytic activity was therefore attributed in part to the CYP2A6 genetic polymorphism. Variation in CYP2A6 activity in this sample group was not associated with gender but, interestingly, it did show an inverse association with plasma ferritin; an indicator of body iron stores. Higher rates of coumarin 7-hydroxylation were found in individuals with low body iron stores (plasma ferritin < 20 microg/l) compared with subjects having normal body iron store status. Subjects (n = 16) with iron overload (plasma ferritin > 300 microg/l) also tended to have elevated rates of coumarin 7-hydroxylation. These results suggest an increased CYP2A6 expression in subjects who have excessive body iron stores. Further investigations into the underlying factors that may lead to increased expression of CYP2A6 in association with abnormal body iron stores are currently in progress in our laboratory. PMID- 11927842 TI - No association between functional catechol O-methyl transferase 1947A>G polymorphism and smoking initiation, persistent smoking or smoking cessation. AB - Nicotine stimulates dopamine release and activates dopaminergic reward neurones in central pathways giving rise to dependence. Catechol O-methyl transferase (COMT) inactivates extraneuronally released dopamine and is present in dopaminergic brain regions. A functional polymorphism (COMT 1947A>G) resulting in increased enzyme activity has been associated with alcoholism and polysubstance abuse. We examined the relationship between the COMT 1947A>G polymorphism and smoking initiation, smoking persistence and smoking cessation. We genotyped 266 current smokers, 270 ex-smokers and 265 lifetime non-smokers (never smokers), matched for age and gender, for the COMT 1947A>G polymorphism. Smoking status was ascertained by self-report. There was no difference in genotype frequencies between never smokers and ever smokers (current + ex-smokers); between non smokers (never + ex-smokers) and current smokers; or between current smokers and ex-smokers. These data suggest that the COMT 1947A>G polymorphism is not associated with smoking initiation, smoking persistence or smoking cessation. PMID- 11927841 TI - Cytochrome P450 2C9 polymorphisms: a comprehensive review of the in-vitro and human data. AB - The discovery of six distinct polymorphisms in the genetic sequence encoding for the cytochrome P450 2C9 (CYP2C9) protein has stimulated numerous investigations in an attempt to characterize their population distribution and metabolic activity. Since the CYP2C9*1, *2 and *3 alleles were discovered first, they have undergone more thorough investigation than the recently identified *4, *5 and *6 alleles. Population distribution data suggest that the variant *2 and *3 alleles are present in approximately 35% of Caucasian individuals; however, these alleles are significantly less prevalent in African-American and Asian populations. In vitro data have consistently demonstrated that the CYP2C9*2 and *3 alleles are associated with significant reductions in intrinsic clearance of a variety of 2C9 substrates compared with CYP2C9*1; however, the degree of these reductions appear to be highly substrate-dependent. In addition, multiple in-vivo investigations and clinical case reports have associated genotypes expressing the CYP2C9*2 and *3 alleles with significant reductions in both the metabolism and daily dose requirements of selected CYP2C9 substrates. Individuals expressing these variant genotypes also appear to be significantly more susceptible to adverse events with the narrow therapeutic index agents warfarin and phenytoin, particularly during the initiation of therapy. These findings have subsequently raised numerous questions regarding the potential clinical utility of genotyping for CYP2C9 prior to initiation of therapy with these agents. However, further clinical investigations evaluating the metabolic consequences in individuals expressing the CYP2C9*2, *3, *4, *5, or *6 alleles are required before large-scale clinical genotyping can be recommended. PMID- 11927843 TI - Intron 3 16 bp duplication polymorphism of p53 is associated with an increased risk for breast cancer by the age of 50 years. AB - We used a large population-based case-control study to determine whether three known p53 polymorphisms, intron 3 16 bp duplication, codon 72(Arg/Pro) and intron 6 MspI restriction fragment length polymorphism, alter the risk for breast cancer in German women. For all three polymorphisms, the odds ratios (ORs) for breast cancer were increased in women carrying the rare allele; however, this was statistically significant only for the 16 bp duplication polymorphism. Compared with the 16 bp duplication wild-type A1/A1 genotype, ORs for A1/A2 genotype and A2/A2 genotype were 1.3 [95% confidence interval (CI) 1.0-1.7] and 1.7 (95% CI 0.8-3.4), suggesting an allele dosage effect (trend test, P = 0.03). Significant evidence was found for a differential effect by family history of breast cancer (P = 0.03 for interaction), with the OR being 5.3 among women with a first degree family history. Our data suggest that inheritance of an intronic polymorphism in the p53 gene increases breast cancer risk appreciably in women by the age of 50 years with a family history of breast cancer in the German population. PMID- 11927844 TI - Management of posterior uveal melanoma: past, present, future. PMID- 11927845 TI - Preclinical and phase 1A clinical evaluation of an anti-VEGF pegylated aptamer (EYE001) for the treatment of exudative age-related macular degeneration. AB - BACKGROUND: Recent studies have suggested that vascular endothelial growth factor (VEGF) is an important stimulus for the growth of new blood vessels in the eye. Anti-VEGF therapy is thus a potential treatment for exudative macular degeneration and diabetic retinopathy. METHODS: Previously described animal models of vascular leakage and ocular neovascularization, including the Miles assay, the rat corneal angiogenesis model, and the mouse retinopathy of prematurity (ROP) model, were used to study this drug. After these studies, a phase IA single ascending dose study of intravitreal injections of the drug was performed in 15 patients with subfoveal choroidal neovascularization secondary to exudative age-related macular degeneration (AMD). RESULTS: The Miles assay model showed almost complete attenuation of VEGF-mediated vascular leakage following addition of EYE001, and the corneal angiogenesis model also showed a significant reduction in neovascularization with EYE001. The ROP model showed inhibition of 80% of the retinal neovascularization compared with controls (P = 0.0001). The phase IA safety study of patients with exudative AMD showed no significant safety issues related to the drug. Ophthalmic evaluation revealed that 80% of patients showed stable or improved vision 3 months after treatment and that 27% of eyes demonstrated a three-line or greater improvement in vision on the Early Treatment for Diabetic Retinopathy Study chart at this time. CONCLUSION: Anti-VEGF therapy is a promising new avenue for the treatment of neovascular diseases of the eye, including exudative macular degeneration and diabetic retinopathy. Preclinical data from studies with EYE001 support clinical evaluation of its efficacy in such diseases. This report is the first to describe administration of anti-VEGF therapy in humans for exudative macular degeneration and shows the safety of such therapy for single injections. Further clinical studies are necessary to determine the safety of multiple intravitreal injections of EYE001 and larger studies are needed to prove the efficacy of this novel, potentially therapeutic agent for neovascular AMD. PMID- 11927846 TI - Management of recurrent retinal detachment in silicone oil-filled eyes. AB - PURPOSE: To report causes of failure, management options, and outcomes after reoperations for recurrent retinal detachment in silicone oil-filled eyes. METHODS: One hundred eighteen silicone oil-filled eyes with recurrent retinal detachment were managed with revision of vitrectomy with membrane surgery with or without silicone oil removal, just scleral buckling, or both. Anatomical success was defined as complete reattachment of the retina, and functional success was defined as recovery of ambulatory visual acuity of >or=5/200 at the last follow up (mean follow-up, 29.7 months). RESULTS: In 82.2% of the cases, proliferative vitreoretinopathy was responsible for recurrent retinal detachment in silicone oil-filled eyes. Reoperations without removal of the silicone oil were performed in 65.3% of the cases. Anatomical success occurred in 62.7% of the eyes, and functional success occurred in 52.5%. Silicone oil was removed in 59.5% of the eyes with retinal reattachment; the retina remained attached in 90.9% of the eyes. Predictors of poor anatomical success were presence of posterior diffuse proliferative vitreoretinopathy and combined posterior and anterior proliferative vitreoretinopathy (P <0.02). CONCLUSION: Reoperations for recurrent retinal detachment in silicone oil-filled eyes were successful in nearly two thirds of the cases, and over one half of the eyes recovered ambulatory vision. PMID- 11927847 TI - Phacoemulsification combined with silicone oil removal through the posterior capsulorhexis tear. AB - PURPOSE: To evaluate phacoemulsification combined with silicone oil removal through the posterior capsulorhexis tear. METHOD: The records of 20 patients with cataract after silicone oil repair of retinal detachment with giant retinal tear or complicated by proliferative vitreoretinopathy were reviewed. All eyes underwent phacoemulsification, posterior capsulorhexis, aspiration of silicone oil with an irrigation-aspiration hand piece, and intraocular lens implantation into the capsular bag. Lens calculation was based on the fellow eye. The mean follow-up period after cataract surgery was 15.4 months. RESULTS: There were three recurrences of retinal detachment (15%). Transient corneal edema was noted in two eyes. Unpredictable refraction was the main problem, but the myopic refractive error was <4.5 diopters in all cases. CONCLUSION: Cataract extraction combined with silicone oil removal is an effective technique. PMID- 11927848 TI - Determination of the solubility of perfluorocarbon liquids in silicone oil in vitro and in vivo. AB - PURPOSE: To investigate the solubility of perfluorocarbon liquids (PFCL) in silicone oil. METHODS: Forty-eight samples of silicone oil (1,300 mPas, n = 22; 5,000 mPas, n = 26) were analyzed for dissolved fluorocarbon molecules after surgical removal from patients who had initially undergone vitreoretinal surgery with (n = 41) and as control without (n = 7) the use of perfluorodecalin in headspace gas chromatography. In vitro, the solubility of three different PFCL perfluorooctane (PFO), perfluorodecalin (PFD), and fluoromethylcyclohexane (FMCH) in silicone oil of various viscosities was determined. The diffusion phenomena during a direct exchange were studied. RESULTS: In 39 of 41 silicone oil samples removed from patients who had undergone vitreoretinal surgery with the use of PFD, small amounts of dissolved perfluorocarbons could be detected. The mean value in 5,000-mPas silicone oil was 939.0 x 10-4 m/% and in 1,300-mPas silicone oil was 322.75 x10(-4) m/%. No perfluorocarbon molecules were found in seven control patients. In vitro, the following maximum solubilities in 1,000-mPas silicone oil were measured at room temperature: PFO, 3.2 m/%; PFD, 5.1 m/%; and FMCH, 10.3 m/%. The maximum values measured in 5,000-mPas silicone oil were PFO, 3.3 m/%; PFD, 5.7 m/%; and FMCH, 8.5 m/%; and in 100-mPas silicone oil were PFO, 2.4 m/%, and PFD, 5.1 m/%. CONCLUSION: Perfluorocarbon liquids dissolve in silicone oil. This may lead to transient formation of "heavy silicone oil," but no stable heavy silicone oil can be created adding PFCL. Intraocularly, retained PFCL vanish in silicone oil and are removed during silicone oil removal. PMID- 11927849 TI - Vitreoretinal management of posteriorly dislocated plate haptic silicone intraocular lenses. AB - PURPOSE: To report the vitreoretinal management of posteriorly dislocated plate haptic silicone intraocular lenses (PHSIOLs) and to present a surgical algorithm for this problem. METHODS: A retrospective analysis of 12 cases of immediate and delayed posterior dislocation of a PHSIOL after phacoemulsification was performed. RESULTS: The ages of the patients ranged from 26 years to 82 years (mean, 68 years); six of the patients were male. Delayed posterior dislocation of the PHSIOL occurred in 7 eyes (after Nd:YAG laser capsulotomy in 4 cases), while intraoperative dislocation occurred in 5 eyes. Defects of the anterior capsulorhexis rim or posterior capsule were present in nine eyes. The mean time from PHSIOL dislocation to definitive treatment was 4.8 months (range, 0.25-13 months). All patients underwent pars plana vitrectomy and implant exchange or repositioning. The dislocated PHSIOL was removed through the transpupillary route in 8 cases, while it was segmented and delivered through the pars plana in 3. Two patients underwent implant repositioning in the ciliary sulcus, although subsequent redislocation in one case required transpupillary removal and exchange. The mean follow-up period was 6.5 months (range, 2-18 months), with a final best corrected visual acuity of 6/12 or better in 10 eyes. CONCLUSIONS: Delayed and immediate dislocation of PHSIOL can occur in eyes with a defect in capsular integrity. The implant may be repositioned or exchanged following vitrectomy with either transpupillary or pars plana removal, resulting in generally excellent postoperative visual acuity and minimal complications. PMID- 11927851 TI - Macular hole repair with minimal vitrectomy. AB - PURPOSE: Conventional macular hole surgery usually involves removing 80% of the intraocular contents to repair a hole a few hundred micrometers in diameter. Although the success rate for conventional surgery is good, it may be possible to reduce the number of complications with less invasive surgery. METHODS: A newly designed microspatula knife was used to dissect the connection between the vitreous and the retina previously delineated by optical coherence tomography. The posterior vitreous was not stripped from the retinal surface. Limited vitrectomy over the hole was performed to create a space for a gas bubble. RESULTS: The macular holes in three eyes of three patients were closed with this technique with no operative or postoperative complications after a mean follow-up of 8.7 months. The mean change in visual acuity was 6.3 lines. CONCLUSION: It is possible to repair macular holes by using optical coherence tomography to guide the dissection of the vitreous from the macular hole followed by limited vitrectomy. By using a less invasive approach, it may be possible to repair macular holes in less operative time and with fewer complications. PMID- 11927850 TI - Types of macular holes encountered during diabetic vitrectomy. AB - PURPOSE: To determine the types of macular holes encountered during vitrectomy for complicated cases of diabetic retinopathy. METHODS: This is a retrospective study of consecutive cases of macular holes diagnosed either before or during pars plana vitrectomy for complications of proliferative diabetic retinopathy over a 6-year period. The morphology of these holes is described. Possible mechanisms of their development, surgical considerations, and results after a follow-up of at least 3 months are discussed. RESULTS: Nineteen consecutive cases of macular holes associated with proliferative diabetic retinopathy were reviewed over a 6-year period. Five cases were lamellar and 14 were full thickness. The 14 cases of full thickness macular holes occurred in patients with the following conditions: tractional rhegmatogenous retinal detachment (5); tractional retinal detachment (3); premacular hemorrhage (3); and cystic macular edema (3). Closure of the full thickness holes was achieved in 8 of 11 patients who were followed up for at least 3 months. All the patients had visual improvement, but no patient had a visual acuity of better than 20/100. CONCLUSION: Macular holes may occur in proliferative diabetic retinopathy in different configurations. Full thickness macular holes can be closed in most cases. Functional improvement can be achieved. PMID- 11927852 TI - Indocyanine green and fluorescein angiography in nonarteritic anterior ischemic optic neuropathy. AB - PURPOSE: To assess the value of indocyanine green angiography (ICGA) for demonstrating choroidal vascular abnormalities in patients with nonarteritic anterior ischemic optic neuropathy (NAION). METHODS: The authors compared the ICGA and fluorescein fundus angiography characteristics of peripapillary circulation in 11 patients with acute NAION. There were 7 men and 4 women; the age range for the patients was 36 years to 72 years (mean +/- SD, 47.7 +/- 10.76 years). The angiographic factors considered significant were delay of peripapillary choroidal filling in the vertical watershed zone, leakage from the optic disk, and absolute filling defects on the disk. The authors compared the incidence of a vertical peripapillary watershed zone in the eyes of the 11 patients with that in the normal eyes of 50 controls (age range, 44-79 years) who had unilateral age-related macular degeneration. RESULTS: Indocyanine green angiography revealed a peripapillary watershed zone in 8 of 11 patient eyes and 23 of 50 control eyes. There was no statistical difference in the number of eyes affected in each group (chi2 = 0.53; P = 0.47). Fluorescein fundus angiography showed leakage from the disk in 10 of 11 patients, whereas ICGA highlighted this problem in only 7 of the patients. The choroidal filling time of the watershed zones was significantly longer with ICGA (t = 3.13; P = 0.011). CONCLUSION: Although ICGA allows better visualization of the choroidal watershed zones associated with NAION, it did not reveal any significantly different incidence of vertical choroidal watershed zone encompassing the optic disk for patients with NAION and controls. Fluorescein fundus angiography better visualized leakage from the disk in the patient group. These findings indicate that ICGA offers no significant advantage in terms of clinical diagnosis and management of NAION. PMID- 11927853 TI - Imaging analysis with optical coherence tomography: relevance for submacular surgery in high myopia and in multifocal choroiditis. AB - PURPOSE: To classify the preoperative and postoperative optical coherence tomography (OCT) findings for subfoveal choroidal neovascularization (CNV) related to high myopia and multifocal choroiditis (MFC) and to correlate these findings with surgical outcome. METHODS: Ten consecutive patients presenting with subfoveal CNV related to either MFC or degenerative myopia were evaluated. Each patient underwent a biomicroscopic examination, fluorescein and indocyanine green angiographies, as well as OCT before and after surgical removal of CNV. Four different parameters were considered in the analysis of all OCT scans: tissue reflectivity, location of the CNV band, presence or absence of a separation zone, and reflectivity underneath the retinal pigment epithelium (RPE) band. RESULTS: For all six eyes with MFC, OCT showed a hyperreflective band anterior to the RPE with a separation zone and an optically clear zone underneath the RPE. Visual acuity improved in all six eyes. For the degenerative myopia group (4 eyes), OCT revealed findings similar to those observed for MFC for 1 eye, which had a favorable postoperative outcome. The remaining three myopic eyes with different OCT patterns had poor postoperative outcomes. CONCLUSION: Optical coherence tomography can provide preoperative clues for submacular surgery. Eyes with CNV located anterior to and separated from the RPE that have an "optically clear zone" underneath are the best candidates for surgical removal. Such a feature was correlated with a good postoperative outcome. PMID- 11927854 TI - Comparison of pH-adjusted bupivacaine with a mixture of non-pH-adjusted bupivacaine and lignocaine in primary vitreoretinal surgery. AB - PURPOSE: To compare the efficacy of alkalinized bupivacaine with that of a mixture of nonalkalinized bupivacaine and lignocaine for local anesthesia in primary vitreoretinal surgery. METHODS: Through a prospective, double-blind, randomized, controlled clinical trial, 540 consecutive patients undergoing primary vitreoretinal surgery received either alkalinized 0.5% bupivacaine (group B) or a mixture of nonalkalinized 0.5% bupivacaine and 2% lignocaine (group BL), both of which were administered with hyaluronidase, for periocular anesthetic block. The periocular block involved two injections in the extraconal space-one in the lower temporal quadrant and the other in the medial periconal space. The efficacy of the block was graded from 0 to 5 depending on the adequacy of anesthesia and akinesia and the need for local supplementation. RESULTS: Adequate anesthesia and akinesia (grade 5) were achieved in 72.2% of the patients in group B compared with 57.4% in group BL (P = 0.0003). Intraoperative supplementation was needed in 9.6% and 20.7% of the patients in groups B and BL, respectively (P = 0.0003). Postoperative analgesics were required in 7.4% of the patients in group B and in 15.2% of those in group BL (P = 0.004). CONCLUSION: Alkalinized 0.5% bupivacaine provides better quality of anesthesia than does the mixture of nonalkalinized 0.5% bupivacaine and 2% lignocaine for patients undergoing primary vitreoretinal surgery. PMID- 11927855 TI - Diagnostic and therapeutic challenges. PMID- 11927856 TI - Translumenal Nd:YAG laser embolysis for branch retinal artery occlusion. PMID- 11927857 TI - Anterior segment ischemia after vitrectomy in sickle cell disease. PMID- 11927858 TI - A case of hemophagocytic syndrome with retinal changes resembling acute posterior multifocal placoid pigment epitheliopathy. PMID- 11927859 TI - Melanocytoma of optic disk in a patient with neurofibromatosis 2. PMID- 11927860 TI - Bull's eye maculopathy in cat-eye syndrome. PMID- 11927862 TI - Treatment of angiomatous lesions of the retina with photodynamic therapy. PMID- 11927861 TI - A case of a five-year-old boy with acute retinal necrosis. PMID- 11927863 TI - A new and simple method for performing vitreous lavage. PMID- 11927864 TI - Elevated macular retinoschisis associated with Goldmann-Favre syndrome successfully treated with grid laser photocoagulation. PMID- 11927865 TI - Spontaneous disappearance of presumed retinal astrocytic hyperplasia. PMID- 11927867 TI - Transient amaurosis associated with intraocular gas during ascending high-speed train travel. PMID- 11927866 TI - Papillophlebitis associated with coexisting factor V Leiden and prothrombin G20210A mutations. PMID- 11927868 TI - A new wide-angle endoillumination probe for use during vitrectomy. PMID- 11927869 TI - A compact, inexpensive face-down positioning device. PMID- 11927870 TI - Gonioscopic detection of a ciliary epithelial tear in atopic dermatitis. PMID- 11927871 TI - Violence in the NHS: take precautions. PMID- 11927872 TI - Detecting and managing undernutrition of older people in the community. AB - Undernutrition is an important public health issue which is frequently undetected and untreated. Disease and illness are the major causes of undernutrition in this country, and older people are a particularly vulnerable group. Effective screening is needed to reduce the prevalence of malnutrition in older people and when this is established, action can then be taken to address the problem. Community nurses and the primary care team have a key role to play in making this happen. The recently developed Malnutrition Advisory Group (MAG) nutrition screening tool, for adults will help community staff and primary care staff to identify those older patients most at risk of malnutrition in the community. Routine nutrition screening of vulnerable individuals is the first stage in raising standards of nutrition care. Further improvements can be made through the development of appropriate nutritional guidelines, clinical risk management strategies provides opportunities to highlight the importance of nutrition care of older people. PMID- 11927873 TI - Prescribing analysis and cost tabulation (PACT) data: an introduction. AB - PACT (prescribing analysis and cost tabulation) data is a national data set which analyses prescribing data in terms of cost and number of items (volume). At an organizational level, PACT is used to monitor and control prescribing cost and to set prescribing budgets. At individual practitioner level, it is used as an educational and audit tool. The Prescription Pricing Authority (PPA) has sent general practitioners (GPs) quarterly summary reports of their own prescribing since 1988. From 2000 prescriptions written by nurse prescribers have been added to the national PACT data set, and health authorities, practices and trusts will now be analysing both nurse and GP prescribing quality. Because of differences in the way they work, nurse prescribers will not automatically receive individual reports of their own prescribing. However they should be able to access this information on request through their local health authority prescribing advisers and primary care group pharmacists. Based on GP experience, nurse prescribers should find PACT data a useful learning resource for individual and team practice development. PMID- 11927874 TI - A multidisciplinary education project in primary care. AB - The Department of Health (DoH) document, 'The New NHS - Working Together' (1998b) recognizes that education and training have a crucial role to play in the development of effective interprofessional work among practitioners. The aim of this article is to provide an account of a project that has been implemented in an East London National Health Service (NHS) Primary Health Trust as well as the experience of using practice-based shared learning with a group of clinical nurse specialists and other health professionals. It outlines the beginnings of the development of the group and discusses the process of facilitating learning, which would enable practitioners to develop in-house training for their staff in clinical practice throughout the Trust. Evaluation of the experience and reflection on the learning that occurred highlighted both positive and negative outcomes at individual and group levels. Findings revealed that shared learning included wider appreciation of each other's skills and increased confidence in multidisciplinary working. Disadvantages, however, included shared learning being considered time-consuming and difficult to facilitate, with participants struggling to cover their normal workloads as well as attending the training programme. PMID- 11927875 TI - District nurses and home carers 3: project evaluation. AB - This article, the last in a series of three, discusses the evaluation of a project established to provide a programme of education for social services' home carers that focused on the development of skills relating to care work. This education was delivered by district nurses (DNs) to address a number of problems associated with the provision of personal care identified by the home carers and their DN colleagues. An action research framework was used to improve collaborative working between the staff of the two disciplines. The article outlines some of the key findings from the evaluation of the project in relation to the structure, the process and the outcome of the project. The project evaluation was mainly positive and this success can be partly attributed to the fact that an action research method was used to drive the project. PMID- 11927876 TI - Urinary sheaths: assessment, prescription and evaluation. AB - Urinary sheaths are a common solution to male urinary incontinence, but can be a source of much anxiety and discomfort if incorrectly fitted. This article discusses the process of assessment that must precede the prescription of a urinary sheath system. Assessment of bladder function and correct fitting and sizing are essential. The article also discusses the range of urinary sheath systems available, some of the problems that may arise in their use, and the processes of evaluation that should accompany use. PMID- 11927877 TI - The concept of priority as it relates to a community mental health team. AB - This article illustrates how a concept clarification exercise can provide evidence to inform local policy development. Based on the framework developed in Walker and Avant's (1995) concept clarification theory, the concept of priority is examined in the context of a team of community mental health nurses. Themes of risk, multidisciplinary working, resources, and nursing role are identified as key areas for consideration, and the difficulties existing between government policy directives towards the severely mentally ill and health promotion are discussed. The article highlights how nursing theory can integrate the concept analysis of priority in planning client care. This is presented using Peplau's (1952) model as a guide to further enhance meaning of generated priorities to nursing practice. PMID- 11927878 TI - Learning disabilities and equity of access. PMID- 11927879 TI - Complaints about nurses are on the increase. PMID- 11927880 TI - A National Patient Safety Agency to be launched. PMID- 11927881 TI - Patient who died after being neglected during the night. PMID- 11927882 TI - Health Service Ombudsman report: decline in quality of nursing care? AB - A number of cases of failures in nursing care have been revealed in the recent Health Service Ombudsman (HSO) Annual Report for England 2000-01. This article highlights some cases from the report as well as recent HSO investigations carried out over the past 4 months. PMID- 11927883 TI - Dysphagia: the management and detection of a disabling problem. AB - Dysphagia represents a varying group of swallowing difficulties commonly encountered in patients in both acute and community settings. It accompanies a variety of disease states, can be neuromuscular or mechanical/obstructive in origin and encompasses varied prognoses and outcomes. Its consequences include dehydration, malnutrition, bronchospasm, airways obstruction, aspiration pneumonia and chronic chest infection, social isolation, depression and detrimental psychosocial effects. Current "best evidence" in screening, assessment and management is of variable quality but demonstrates that nurses have an important role to play in interventions entailing multiprofessional collaboration within individually tailored programmes. Clear benefits for patients have been indicated. There are gaps in the knowledge base, especially in relation to psychosocial effects and treatment strategies and the nursing contribution in this area. PMID- 11927884 TI - Causes and effects of chronic obstructive pulmonary disease. AB - Chronic obstructive pulmonary disease (COPD) is a chronic, slowly progressive disease which is a significant cause of disability and death (British Thoracic Society (BTS), 1997). Despite the clinical and economic burden it creates, it is a relatively poorly understood disease, with many of those with COPD being undiagnosed. This article will discuss some of the causes and effects of COPD and give particular consideration to its impact on patients. It will also consider how the impact of the disease may be reduced, giving greater attention to two specific interventions, namely pulmonary rehabilitation and smoking cessation. The BTS guidelines as a framework for the management of COPD, and the need for a collaborative approach between primary and secondary care, will also be highlighted. PMID- 11927885 TI - Perceptions of older people who have experienced physical restraint. AB - It is well documented that the use of physical restraints on older people has been linked to negative clinical outcomes. However, less is known about the personal perspective of those who have been restrained. This study examines the perceptions of older people who have experienced physical restraints in a rehabilitation ward. A purposive sample was used of 17 male and female patients who were restrained. The patients were interviewed using the Subjective Experience of Being Restrained instrument (Strumpf and Evans, 1988) which is a semi-structured interview schedule. The most commonly used restraint devices included side rails, screw-on tabletops and reclining chairs. The data were analysed using content analysis. The results indicate mixed feelings regarding physical restraints. Patients' impressions of physical restraints included indifference of the devices to their perceived safety value. Overall, a minority of patients (n = 4) had positive feelings about physical restraints as they provided a sense of security to them. However, the negative comments of the patients were more prevalent and their responses were categorized in terms of institutional control, ritualised care, entrapment and discomfort, and possible alternatives. PMID- 11927886 TI - Breakthrough pain: assessment and management in cancer patients. AB - This article provides an overview of breakthrough pain in cancer patients, what causes it, current treatment options and the impact it has on individuals. It considers the importance of accurate assessment, the use of assessment tools and the growing role of nurses in managing this challenging pain syndrome. The article aims to open the debate on the need for new choices in pain management. While many advances have been made in the treatment of pain, there is still room for improvement in both the pharmaceutical and general management of the condition. Suggestions are made as to how these may be met. PMID- 11927887 TI - Care study: the effect of pain on a patient with leg ulcers. AB - The following care study aims to explore the effects of wound-related pain on a patient with leg ulceration. The study illustrates current deficits within the author's clinical area, a day hospital for elderly people, in relation to how both a patient's pain and quality of life are measured. It suggests means by which this could be overcome, e.g. by the use of appropriate pain and quality of life pain measurement scales. The case study also highlights the hospital community divide, with reference to communication and the exchange of information relating to a patient's pain. It advocates optimizing interdisciplinary team communication and discusses how developing closer working relationships and improving education can play a part in enhancing communication. While this case study is concerned with an individual patient, it aims to demonstrate that the problems identified and the resulting indications for clinical practice will promote an increasingly holistic view of the patient and benefit all future patients with leg ulceration. PMID- 11927888 TI - Legal aspects of consent 9: when parents are overruled. PMID- 11927889 TI - The Theracute alternating pressure replacement mattress. AB - The use of alternating pressure mattresses is still acknowledged as an effective adjunct in the prevention and treatment of pressure damage; however, with the plethora of equipment now available, there is an increasing need to provide information that supports the practitioners' decisions ensuring individual patients' clinical needs are met. This article reviews a new addition to the Talley Medical range of alternating pressure mattresses, the Theracute mattress replacement system. PMID- 11927890 TI - Can we standardize titles and levels in nursing? PMID- 11927891 TI - Nurse-led walk-in centre model needs rethinking. PMID- 11927892 TI - Large patients must be treated with respect. PMID- 11927893 TI - Learning disability nurse who learnt from her mistake. PMID- 11927894 TI - Pain management 1: psychological and social aspects of pain. AB - This two-part article presents psychological and social factors which affect pain perception and response and the implications of these for nursing practice. In this article, the complex interactions between neurophysiological and psychological factors are outlined and theories of pain perception and ways in which the pain experience can be modulated are presented. The role of psychological factors, attitudes, beliefs and expectations of both patient and practitioner, pain behaviours and ability to cope are discussed. In the second part, these are further elaborated with particular reference to the nurse-patient interaction. The use of psychological approaches to augment clinical practice, such as education, reduction of anxiety and improving coping ability, are suggested. Finally, the importance of communication skills in pain management is addressed. PMID- 11927895 TI - Factors affecting how patients sleep in the hospital environment. AB - Promoting an environment conductive to sleep in hospital is a challenge. Although patients may appear to sleep in hospital, it may not be refreshing or restorative. The reasons for this can be categorized into three groups: environmental, physiological and psychological. These factors can work concomitantly, making sleep virtually impossible for some patients. Nurses can help their patients by understanding what influences sleep patterns and acting on this knowledge. This may include a variety of interventions, from allowing patients to carry out their own bedtime routine, to explaining to elderly people how their sleeping patterns changes with age. PMID- 11927896 TI - Legal aspects of consent 10: determining "best interests". PMID- 11927897 TI - Sexuality: meaning and relevance to learning disability nurses. AB - The author posits the view that, while there has been considerable growth in the quantity of published material on the subject of sexuality, few authors have attempted to define what is meant by "sexuality". It is suggested that because the concept is intangible and ill defined many nurses find it difficult to approach this aspect of holistic care. In an attempt to address this problem, the main body of this article seeks to explore the concept of sexuality and in so doing seeks to provide the reader with clarification as to what the concept entails. The latter part of the article considers the concept of sexuality in relation to the author's client group, namely persons with learning disabilities. In doing so, the author seeks to demonstrate the relevance of the subject to nurses working with people with learning disabilities. PMID- 11927898 TI - Caring for adults with a learning disability in the community. AB - A recurring theme within the literature is gaps in the delivery of services for people with a learning disability living in the community. These gaps occur between health professionals, primary and secondary care and specialisms within nursing. Gaps also exist between social service policy and implementation, health and social care. Recent national and local reports have sought to address these issues by promoting ways for health and social services to work in partnership. The theme of health promotion is highlighted in all these documents, but implementation of health promotion has been firmly placed within the boundaries of primary care. This poses a dilemma for primary care: does the primary care team or a more specialist community learning disability team provide better care for people with a learning disability? This article summarizes strategies and policies within the literature and difficulties that need to be considered when offering a service to people with a learning disability. PMID- 11927899 TI - Devising an effective general nursing continence assessment tool. AB - Incontinence assessment forms are routinely used by nurses in order to implement the appropriate management, treatment and referral of patients suffering from bladder dysfunction. A thorough literature search using recognized databases revealed that no specific work has been carried out in respect of validating symptom scoring assessment tools for nurses. Validated symptom scores are available but these are generally used within specific areas of medicine, e.g. prostatic screening incorporates lower urinary tract symptom scores, and physiotherapy, to measure pelvic floor tone. This article examines the need for validity and symptom scoring within a general nursing incontinence assessment tool. PMID- 11927900 TI - Opsite Plus in the managment of low to moderate exuding wounds. AB - This article discusses the use of OpSite Plus (formerly known as OpSite Post-Op) in the community setting. This semipermeable film dressing with an absorbent pad was evaluated by the community hospital and by practice and district nurses within Plymouth's NHS Primary Care Trust. PMID- 11927901 TI - Healthcare professions must work better as teams. PMID- 11927902 TI - Extracorporeal blood detoxification by sorbents in treatment of hepatic encephalopathy. AB - Extracorporeal blood detoxification by sorbent therapy long has been applied in treatment of hepatic failure and encephalopathy, starting with hemoperfusion columns and more recently with the currently marketed Liver Dialysis Unit. Liver Dialysis employs hemodiabsorption (dialysis of blood against powdered sorbents including charcoal and cation exchanger) to remove selectively numerous small molecular-weight toxins of hepatic failure. Liver Dialysis is used in treatment of acute hepatic encephalopathy (AHE) because of decompensation of chronic liver disease (A-on-C) or fulminant hepatic failure (FHF). Controlled, prospective and randomized studies of daily 6-hour Liver Dialysis have shown physiologic and neurologic improvement of patients with AHE, regardless of etiology. Liver dialysis significantly improved the incidence of positive outcomes (recovery of hepatic function or improvement for transplant) of A-on-C patients versus controls (71.5% treated, and 35.7% control, P =.036), but had an insignificant improvement in outcome of patients with FHF as compared with the control group. Other extracorporeal sorbent devices are now in clinical testing phase. The molecular adsorbent regenerating system (MARS) device employs a polysulfone high permeability dialyzer with albumin on the dialysate side to aid transfer of protein-bound toxins such as bilirubin and bile acids across the membranes. Sorbent columns of charcoal and an anion exchanger remove hepatic toxins from the albumin dialysate, and a second dialyzer removes water-soluble toxins, such as ammonium. Clinical results of daily MARS treatments of patients with hepatic failure are similar to that of Liver Dialysis, with neurologic and physical improvement occurs in most patients with AHE, and improved outcome for patients with A-on-C. The system extends the life of patients with hepatorenal syndrome. PF-Liver Dialysis is an experimental device combining hemodiabsorption with push pull sorbent-based pheresis with powdered sorbent surrounding plasmafilters. PF Liver Dialysis (Hemocleanse, Inc, W. Lafayette, IN) has been tested in a few patients with hepatic failure, grade 3-4 encephalopathy, and respiratory and kidney insufficiency. Treatments appeared to be safe and resulted in marked decreases in plasma levels of bilirubin, aromatic amino acids, ammonium, creatinine, and interleukin-1beta (IL-1beta). The PF add-on module adds the capability to Liver Dialysis to remove bilirubin, bile acids, and other strongly protein-bound toxins from treated patients and may be of clinical benefit in management of patients with the most severe hepatic failure and encephalopathy, including patients with FHF or concomitant sepsis. PMID- 11927903 TI - Sorbent hemoperfusion in end-stage renal disease: an in-depth review. AB - The excessive mortality in dialysis patients has rekindled interest in research of adsorbent removal of nontraditional uremic toxins. Middle-molecular-weight substances, predominantly small proteins, have been correlated with specific uremic syndromes and implicated in the uremic state. New developments in polymer technology and carbon pyrrolization techniques have produced sorbents possessing mesopores of sufficient size to trap middle-molecular-weight substances. Clinical application of hemoperfusion devices containing these sorbents is early in its development. Studies related to hemoperfusion in uremia are discussed in detail. PMID- 11927904 TI - Dialysis and hemoperfusion in poisoning. AB - The treatment of serious drug or chemical poisoning relies mostly on the use of standard intensive care measures. Supplementation of intensive care with dialysis and hemoperfusion may be necessary at times to remove the intoxicant in seriously ill patients. Factors governing drug and chemical removal by dialysis and hemoperfusion are discussed, and guidelines given when to employ these techniques. Tables of drugs removed by both techniques are given for guidance. These tables will be updated regularly on the Advances in Renal Replacement Therapy website. PMID- 11927905 TI - Treatment of severe tricyclic antidepressant overdose with extracorporeal sorbent detoxification. AB - Tricyclic overdose can be a medical emergency, and therapy with intravenous bicarbonate is not always successful in preventing cardiac toxicity or coma. Mortality in patients developing these complications is from 1% to 15%. Extracorporeal detoxification with sorbents has been used in treatment of patients with very high drug levels and declining clinical condition. Ten patients with serious drug overdose caused by tricyclics failed to respond quickly to standard therapy and were in stage 3-4 encephalopathy. Nine of these patients were on respirator support, 5 had hypotension, and 6 had QRS widening. Average level was 1,423 microg/L at presentation. Enteral activated charcoal and intravenous (IV) bicarbonate were initiated in the emergency room. The patients were treated for 3 to 4 hours with the Liver Dialysis Unit, a hemodiabsorption device using a cellulosic plate dialyzer and sorbent suspension as dialysate. Inflow and outflow blood levels indicated that the hemodetoxifier removed modest amounts of the tricyclics, metabolites, and other consumed drugs. The clinical improvement of the patients was dramatic, with patients reaching stage 0 or 1 encephalopathy during the treatment. Ventilator support was removed at the end of treatment for 3 patients who had not already developed pneumonia, and for others was prolonged up to 48 hours because of pneumonia, rather than mental status. Average length of stay in the intensive care unit (ICU) was 4.8 days (range 1 to 7 days). None of the patients died despite their high risk for ventricular arrhythmias, seizures, and death. Clinical improvement may have been attributable to removal of free drug from the blood or to removal of drug metabolites. PMID- 11927906 TI - Treatment of acetaminophen-induced hepatitis and fulminant hepatic failure with extracorporeal sorbent-based devices. AB - When a patient with acetaminophen overdose arrives in the emergency room more than 14 hours after ingestion, the value of N-acetylcysteine is unproven and patient mortality is at least 10%. Anecdotal case reports have indicated benefit of extracorporeal detoxification of these late-arriving patients with acetaminophen overdose. We identified 10 patients with serious acetaminophen overdose, 8 that arrived in the emergency room 16 to 44 hours after acetaminophen overdose with plasma levels predicting severe hepatic toxicity, and 2 that arrived in the emergency room 8 to 12 hours after overdose but with exceedingly high levels. All patients developed severe hepatitis (mean peak alanine aminotransferase, 4,052; mean peak protime, 25 seconds). At 16 to 68 hours after overdose, the patients were treated for 4 to 6 hours with the Liver Dialysis System (Hemocleanse Inc, W. Lafayette, IN), a single-access hemodiabsorption system indicated for treatment of serious drug overdose and for treatment of hepatic encephalopathy. Acetaminophen levels fell an average of 73% during treatment. Treatment was repeated 24 or 48 hours later if acetaminophen was still measurable in plasma. All 10 patients recovered intrinsic liver function and general health, with liver enzymes starting to normalize 24 hours after treatment, and were discharged 3 to 7 days after overdose. No patient required liver transplant. Because market introduction of Liver Dialysis, there have been 40 more patients with acetaminophen-induced hepatotoxicity treated with Liver Dialysis. All have recovered liver function without long-term sequelae. Though most of these patients with already established hepatic toxicity from acetaminophen would recover without extracorporeal blood therapy, treatment with the Liver Dialysis System should assure recovery from acute hepatic failure, and may shorten the clinical course of the illness. PMID- 11927907 TI - Development of a culturally sensitive, locality-based program to increase kidney donation. AB - Diabetic kidney disease has become an epidemic in American Indian communities. In Central Arizona, the Salt River Pima-Maricopa, Gila River, and Ak-Chin Indian communities have the highest incidence rate of diabetes in the world. Over 200 diabetic people in these communities receive dialysis because of kidney disease, a frequency that is 10 to 15 times the national average. Very few of these patients ever receive a kidney transplant. The communities formed the Organ Donation and Kidney Transplant Education Committee, which designed culturally sensitive educational materials and trained tribal members to disseminate the information. The goals were to educate tribal members about the choices they have when faced with kidney failure and to increase organ donation within the communities. The committee also produced a 10- to 15-minute educational video presentation for use at community district meetings, health fairs, and local health clinics in the communities. One unexpected early outcome has been the opportunity to reflect on some assumptions and stereotypes about American Indians and organ donation commonly held by health care providers. The local organ procurement organization plans to use the video in training hospital donor requestors. What difference would it make to educate the communities and not change the assumptions of those in the hospitals requesting donation? PMID- 11927908 TI - Medical waste production and disposal arising from renal replacement therapy. AB - The treatment of end stage renal disease (ESRD) makes extensive use of pre sterilised disposable items that are contaminated by blood or other body fluids following use. Commonly these items are incinerated. Because they contain substantial amounts of polyvinylchloride (PVC), their incineration releases polychlorinated dibenzo p dioxins (PCDD) and polychlorinated di benzo p furans (PCF), as well as heavy metals. A consequence of the release of these substances has been the introduction of legislation controlling waste disposal. These issues are likely to impact on the management of waste at the healthcare facility level. In parallel, new PVC-free materials for use in renal replacement therapies have been developed whose incineration is less controversial environmentally. PMID- 11927909 TI - Emergency dialysis in neonatal metabolic crises. AB - Metabolic crises in children with inborn errors of metabolism are caused by the accumulation of neurotoxic metabolites. Rapid elimination of these metabolites is apparently crucial to prevent irreversible neuronal damage; long-term outcome is correlated with the rate of toxin removal during the crisis. The usefulness of different blood purification techniques for removing accumulated neurotoxins is reviewed in this manuscript. PMID- 11927910 TI - Tracing lymphopoiesis with the aid of a pTalpha-controlled reporter gene. AB - A transgenic reporter mouse strain, which expressed the human CD25 (hCD25) surface marker as a reporter under the control of the pre-T cell receptor alpha(pTalpha) promoter, was used to identify lymphoid precursors that expressed pTalpha intracellularly. The hCD25 reporter marked intra- and extrathymic precursors of lymphocytes but not myeloid cells. The earliest intrathymic precursors were CD4(lo)CD8(-)CD25(-)CD44(+)c-Kit(+) cells that expressed elevated levels of Notch-1 mRNA. Clonogenic assays showed that the extrathymic precursors were common lymphoid progenitors (CLPs) that included CD19(-), B220(+), Thy1(+) and CD4(+) cells. Thus, the pTalpha reporter can be used to trace lymphopoiesis between CLPs and alphabeta T cells. The slower extinction of the hCD25 reporter compared to pTalpha enabled us to define points at which pTalpha(-) lineages branched off. PMID- 11927912 TI - Mind your heart. PMID- 11927911 TI - A critical role for the cytoplasmic tail of pTalpha in T lymphocyte development. AB - Signals that emanate from the pre-T cell receptor (pre-TCR) regulate multiple processes required for the development of the alphabeta T cell lineage. In contrast to the gammadelta TCR, the pre-TCR localizes cell-autonomously to membrane rafts, where it appears to signal in a constitutive and ligand independent manner. We addressed here the role played by structural features specific to the cytoplasmic domain of the pre-TCRalpha chain (pTalpha). More specifically, we examined a COOH-terminal proline-rich sequence that might play a role in signal transduction and a juxtamembrane cysteine residue that could be a target for palmitoylation, thus allowing spontaneous raft localization. Expression of pTalpha mutants in transgenic mice, retrovirally transduced T cell precursors and cell lines showed that the pTalpha cytoplasmic tail, in particular the proline-rich domain, plays a crucial role in pre-TCR signaling and T cell development. In contrast, the pTalpha juxtamembrane cysteine appeared to be dispensable for pre-TCR function. PMID- 11927913 TI - Does neutrophil CD38 have a role in Ca++ signaling triggered by beta2 integrin? PMID- 11927915 TI - Dyslipidemia due to retroviral protease inhibitors. PMID- 11927917 TI - Will Zerhouni become the new NIH director? PMID- 11927918 TI - Poor sales trigger vaccine withdrawal. PMID- 11927920 TI - Ebola: small, but real progress. PMID- 11927921 TI - Charity's first move is drug investment. PMID- 11927923 TI - New regulations give UK the lead in stem cell work. PMID- 11927924 TI - Systems biology--the new R&D buzzword? PMID- 11927925 TI - Nature Medicine/UCSD/Salk conference rewards physician-scientists. PMID- 11927926 TI - Barry Bloom. PMID- 11927927 TI - CD4+ T-cell depletion in HIV infection: are we closer to understanding the cause? PMID- 11927930 TI - New anti-HSV therapeutics target the helicase-primase complex. PMID- 11927931 TI - Polygenic or pollyanna? PMID- 11927933 TI - Juvenile autoimmune diabetes: a pathogenic role for maternal antibodies? PMID- 11927934 TI - Antibody gene therapy: old wine in a new bottle. PMID- 11927935 TI - Block the FAS, lose the fat. PMID- 11927936 TI - The skinny on CD39 in immunity and inflammation. PMID- 11927937 TI - Not just research tools--proteasome inhibitors offer therapeutic promise. PMID- 11927939 TI - Gene therapy for cancer using single-chain Fv fragments specific for 4-1BB. AB - Monoclonal antibodies against the T-cell activation molecule 4-1BB have been effective in the treatment of established mouse tumors. To create a vaccine that stimulates the immune system similarly to the efficacious monoclonal anti-4-1BB antibody, 1D8, we constructed a vector encoding cell-bound single-chain Fv fragments from 1D8. We transfected the vector into cells from the K1735 melanoma, selected because of its low immunogenicity and very low expression of major histocompatibility complex class I. The transfected cells induced a strong type 1 T-helper cell response, for which CD4+ but not CD8+ T lymphocytes were necessary and that involved natural killer cells. Vaccinated mice rejected established wild type K1735 tumors growing as subcutaneous nodules or in the lung. An analogous approach may be effective against micrometastases in human patients, including tumors whose expression of major histocompatibility complex class I is very low. PMID- 11927940 TI - Inducer-stimulated Fas targets activated endothelium for destruction by anti angiogenic thrombospondin-1 and pigment epithelium-derived factor. AB - Natural inhibitors of angiogenesis are able to block pathological neovascularization without harming the preexisting vasculature. Here we show that two such inhibitors, thrombospondin-1 and pigment epithelium-derived factor, derive specificity for remodeling vessels from their dependence on Fas/Fas ligand (FasL)-mediated apoptosis to block angiogenesis. Both inhibitors upregulated FasL on endothelial cells. Expression of the essential partner of FasL, Fas/CD95 receptor, was low on quiescent endothelial cells and vessels but greatly enhanced by inducers of angiogenesis, thereby specifically sensitizing the stimulated cells to apoptosis by inhibitor-generated FasL. The anti-angiogenic activity of thrombospondin-1 and pigment epithelium-derived factor both in vitro and in vivo was dependent on this dual induction of Fas and FasL and the resulting apoptosis. This example of cooperation between pro- and anti-angiogenic factors in the inhibition of angiogenesis provides one explanation for the ability of inhibitors to select remodeling capillaries for destruction. PMID- 11927941 TI - CD39 is the dominant Langerhans cell-associated ecto-NTPDase: modulatory roles in inflammation and immune responsiveness. AB - CD39, the endothelial ecto-nucleoside triphosphate diphosphohydrolase (NTPDase), regulates vascular inflammation and thrombosis by hydrolyzing ATP and ADP. Although ecto-NTPDase activities have been used as a marker of epidermal dendritic cells (DCs) known as Langerhans cells, the identity and function of these activities remain unknown. Here we report that Langerhans cells in CD39-/- mice express no detectable ecto-NTPDase activity. Irritant chemicals triggered rapid ATP and ADP release from keratinocytes and caused exacerbated skin inflammation in CD39-/- mice. Paradoxically, T cell-mediated allergic contact hypersensitivity was severely attenuated in CD39-/- mice. As to mechanisms, T cells increased pericellular ATP concentrations upon activation, and CD39-/- DCs showed ATP unresponsiveness (secondary to P2-receptor desensitization) and impaired antigen-presenting capacity. Our results show opposing outcomes of CD39 deficiency in irritant versus allergic contact dermatitis, reflecting its diverse roles in regulating extracellular nucleotide-mediated signaling in inflammatory responses to environmental insults and DC-T cell communication in antigen presentation. PMID- 11927942 TI - Efomycine M, a new specific inhibitor of selectin, impairs leukocyte adhesion and alleviates cutaneous inflammation. AB - Specific interference with molecular mechanisms guiding tissue localization of leukocytes may be of great utility for selective immunosuppressive therapies. We have discovered and characterized efomycines, a new family of selective small molecule inhibitors of selectin functions. Members of this family significantly inhibited leukocyte adhesion in vitro. Efomycine M, which was nontoxic and showed the most selective inhibitory effects on selectin-mediated leukocyte-endothelial adhesion in vitro, significantly diminished rolling in mouse ear venules in vivo as seen by intravital microscopy. In addition, efomycine M alleviated cutaneous inflammation in two complementary mouse models of psoriasis, one of the most common chronic inflammatory skin disorders. Molecular modeling demonstrated a spatial conformation of efomycines mimicking naturally occurring selectin ligands. Efomycine M might be efficacious in the treatment of human inflammatory disorders through a similar mechanism. PMID- 11927943 TI - Complement component C3 promotes T-cell priming and lung migration to control acute influenza virus infection. AB - The complement cascade defines an important link between the innate and the specific immune system. Here we show that mice deficient for the third component of complement (C3-/- mice) are highly susceptible to primary infection with influenza virus. C3-/- mice showed delayed viral clearance and increased viral titers in lung, whereas mice deficient for complement receptors CR1 and CR2 (Cr2 /- mice) cleared the infection normally. Priming of T-helper cells and cytotoxic T cells (CTLs) in lung-draining lymph nodes was reduced, and the recruitment into the lung of virus-specific CD4+ and CD8+ effector T cells producing interferon gamma was severely impaired in C3-/- but not in Cr2-/- mice. Consequently, T helper cell-dependent IgG responses were reduced in C3-/- mice but remained intact in Cr2-/- mice. These results demonstrate that complement induces specific immunity by promoting T-cell responses. PMID- 11927944 TI - Memory CD8+ T cells vary in differentiation phenotype in different persistent virus infections. AB - The viruses HIV-1, Epstein-Barr virus (EBV), cytomegalovirus (CMV) and hepatitis C virus (HCV) are characterized by the establishment of lifelong infection in the human host, where their replication is thought to be tightly controlled by virus specific CD8+ T cells. Here we present detailed studies of the differentiation phenotype of these cells, which can be separated into three distinct subsets based on expression of the costimulatory receptors CD28 and CD27. Whereas CD8+ T cells specific for HIV, EBV and HCV exhibit similar characteristics during primary infection, there are significant enrichments at different stages of cellular differentiation in the chronic phase of persistent infection according to the viral specificity, which suggests that distinct memory T-cell populations are established in different virus infections. These findings challenge the current definitions of memory and effector subsets in humans, and suggest that ascribing effector and memory functions to subsets with different differentiation phenotypes is no longer appropriate. PMID- 11927945 TI - Herpes simplex virus helicase-primase inhibitors are active in animal models of human disease. AB - Herpes simplex virus infections are the cause of significant morbidity, and currently used therapeutics are largely based on modified nucleoside analogs that inhibit viral DNA polymerase function. To target this disease in a new way, we have identified and optimized selective thiazolylphenyl-containing inhibitors of the herpes simplex virus (HSV) helicase-primase enzyme. The most potent compounds inhibited the helicase, the primase and the DNA-dependent ATPase activities of the enzyme with IC50 (50% inhibitory concentration) values less than 100 nM. Inhibition of the enzymatic activities was through stabilization of the interaction between the helicase-primase and DNA substrates, preventing the progression through helicase or primase catalytic cycles. Helicase-primase inhibitors also prevented viral replication as demonstrated in viral growth assays. One compound, BILS 179 BS, displayed an EC50 (effective concentration inhibiting viral growth by 50%) of 27 nM against viral growth with a selectivity index greater than 2,000. Antiviral activity was also demonstrated for multiple strains of HSV, including strains resistant to nucleoside-based therapies. Most importantly, BILS 179 BS was orally active against HSV infections in murine models of HSV-1 and HSV-2 disease and more effective than acyclovir when the treatment frequency per day was reduced or when initiation of treatment was delayed up to 65 hours after infection. These studies validate the use of helicase-primase inhibitors for the treatment of acute herpesvirus infections and provide new lead compounds for optimization and design of superior anti-HSV agents. PMID- 11927946 TI - New helicase-primase inhibitors as drug candidates for the treatment of herpes simplex disease. AB - The vast majority of the world population is infected with at least one member of the human herpesvirus family. Herpes simplex virus (HSV) infections are the cause of cold sores and genital herpes as well as life-threatening or sight-impairing disease mainly in immunocompromized patients, pregnant women and newborns. Since the milestone development in the late 1970s of acyclovir (Zovirax), a nucleosidic inhibitor of the herpes DNA polymerase, no new non-nucleosidic anti-herpes drugs have been introduced. Here we report new inhibitors of the HSV helicase-primase with potent in vitro anti-herpes activity, a novel mechanism of action, a low resistance rate and superior efficacy against HSV in animal models. BAY 57-1293 (N-[5-(aminosulfonyl)-4-methyl-1,3-thiazol-2-yl]-N-methyl-2-[4-(2 pyridinyl)phenyl]acetamide), a well-tolerated member of this class of compounds, significantly reduces time to healing, prevents rebound of disease after cessation of treatment and, most importantly, reduces frequency and severity of recurrent disease. Thus, this class of drugs has significant potential for the treatment of HSV disease in humans, including those resistant to current medications. PMID- 11927947 TI - Elimination of maternally transmitted autoantibodies prevents diabetes in nonobese diabetic mice. AB - The influence of maternally transmitted immunoglobulins on the development of autoimmune diabetes mellitus in genetically susceptible human progeny remains unknown. Given the presence of islet beta cell-reactive autoantibodies in prediabetic nonobese diabetic (NOD) mice, we abrogated the maternal transmission of such antibodies in order to assess their influence on the susceptibility of progeny to diabetes. First, we used B cell-deficient NOD mothers to eliminate the transmission of maternal immunoglobulins. In a complementary approach, we used immunoglobulin transgenic NOD mothers to exclude autoreactive specificities from the maternal B-cell repertoire. Finally, we implanted NOD embryos in pseudopregnant mothers of a non-autoimmune strain. The NOD progeny in all three groups were protected from spontaneous diabetes. These findings demonstrate that the maternal transmission of antibodies is a critical environmental parameter influencing the ontogeny of T cell-mediated destruction of islet beta cells in NOD mice. It will be important to definitively determine whether the transmission of maternal autoantibodies in humans affects diabetes progression in susceptible offspring. PMID- 11927948 TI - Hematopoietic stem cells differentiate into vascular cells that participate in the pathogenesis of atherosclerosis. AB - Excessive accumulation of smooth-muscle cells (SMCs) has a key role in the pathogenesis of vascular diseases. It has been assumed that SMCs derived from the outer medial layer migrate, proliferate and synthesize extracellular matrix components on the luminal side of the vessel. Although much effort has been devoted to targeting migration and proliferation of medial SMCs, there is no effective therapy that prevents occlusive vascular remodeling. We show here that in models of post-angioplasty restenosis, graft vasculopathy and hyperlipidemia induced atherosclerosis, bone-marrow cells give rise to most of the SMCs that contribute to arterial remodeling. Notably, purified hematopoietic stem cells differentiate into SMCs in vitro and in vivo. Our findings indicate that somatic stem cells contribute to pathological remodeling of remote organs, and may provide the basis for the development of new therapeutic strategies for vascular diseases through targeting mobilization, homing, differentiation and proliferation of bone marrow-derived vascular progenitor cells. PMID- 11927950 TI - Improved genetic immunization via micromechanical disruption of skin-barrier function and targeted epidermal delivery. AB - Skin is an attractive target for delivery of genetic therapies and vaccines. However, new approaches are needed to access this tissue more effectively. Here, we describe a new delivery technology based on arrays of structurally precise, micron-scale silicon projections, which we term microenhancer arrays (MEAs). In a human clinical study, these devices effectively breached the skin barrier, allowing direct access to the epidermis with minimal associated discomfort and skin irritation. In a mouse model, MEA-based delivery enabled topical gene transfer resulting in reporter gene activity up to 2,800-fold above topical controls. MEA-based delivery enabled topical immunization with naked plasmid DNA, inducing stronger and less variable immune responses than via needle-based injections, and reduced the number of immunizations required for full seroconversion. Together, the results provide the first in vivo use of microfabricated devices to breach the skin barrier and deliver vaccines topically, suggesting significant clinical and practical advantages over existing technologies. PMID- 11927949 TI - In situ dissection of the graft-versus-host activities of cytotoxic T cells specific for minor histocompatibility antigens. AB - Minor histocompatibility antigens (mHags) are immunogenic peptides from polymorphic cellular proteins that induce strong T-cell responses after human leukocyte antigen (HLA)-matched, mHag-mismatched stem-cell transplantation. mHags with broad or limited tissue expression are target antigens for graft-versus-host (GvH) and graft-versus-leukemia (GvL) reactivities. Separation of these activities is crucial for adoptive immunotherapy of leukemia without GvH disease. Therefore, using a skin-explant assay we investigated the in situ activities of cytotoxic T lymphocytes (CTLs) specific for the ubiquitously expressed mHag H-Y and for the hematopoietic-restricted mHags HA-1 and HA-2. H-Y-specific CTLs, visualized by tetrameric HLA-mHag peptide complexes, infiltrated male skin sections within 24 hours, induced severe GvH reactions of grade III-IV and produced high levels of IFN-gamma. In contrast, CTLs specific for the hematopoietic system-specific mHags HA-1 and HA-2 induced no or low GvH reactions above background and produced little or no interferon-gamma, unless the skin sections were preincubated with HA-1/HA-2 synthetic peptides. These results provide the first in situ dissection of GvH effects by mHag-specific CTLs and show that ubiquitously expressed mHags are the prime targets of GvH disease. PMID- 11927952 TI - Structure of the UGAGAU hexaloop that braces Bacillus RNase P for action. AB - Long-range interactions involving the P5.1 hairpin of Bacillus RNase P RNA are thought to form a structural truss to support RNA folding and activity. We determined the structure of this element by NMR and refined the structure using residual dipolar couplings from a sample weakly oriented in a dilute liquid crystalline mixture of polyethylene glycol and hexanol. Dipolar coupling refinement improved the global precision of the structure from 1.5 to 1.2 A (to the mean), revised the bend angle between segments of the P5.1 stem and corroborated the structure of the loop region. The UGAGAU hexaloop of P5.1 contains two stacks of bases on opposite sides of the loop, distinguishing it from GNRA tetraloops. The unusual conformation of the juxtaposed uracil residues within the hexaloop may explain their requirement in transactivation assays. PMID- 11927953 TI - Crystal structure of an RNA tertiary domain essential to HCV IRES-mediated translation initiation. AB - The hepatitis C virus (HCV) internal ribosome entry site (IRES) RNA drives internal initiation of viral protein synthesis during host cell infection. In the tertiary structure of the IRES RNA, two helical junctions create recognition sites for direct binding of the 40S ribosomal subunit and eukaryotic initiation factor 3 (eIF3). The 2.8 A resolution structure of the IIIabc four-way junction, which is critical for binding eIF3, reveals how junction nucleotides interact with an adjacent helix to position regions directly involved in eIF3 recognition. Two of the emergent helices stack to form a nearly continuous A-form duplex, while stacking of the other two helices is interrupted by the insertion of junction residues into the helix minor groove. This distorted stack probably serves as an important recognition surface for the translational machinery. PMID- 11927955 TI - The mystery of the fat clones. PMID- 11927954 TI - A conserved RNA structure within the HCV IRES eIF3-binding site. AB - The hepatitis C virus (HCV) internal ribosome entry site (IRES) is recognized specifically by the small ribosomal subunit and eukaryotic initiation factor 3 (eIF3) before viral translation initiation. Using extensive mutagenesis and structure probing analysis, we show that the eIF3-binding domain of the HCV IRES contains an internal loop structure (loop IIIb) and an adjacent mismatched helix that are important for IRES-dependent initiation of translation. NMR studies reveal a unique three-dimensional structure for this internal loop that is conserved between viral isolates of varying primary sequence in this region. These data indicate that internal loop IIIb may be an attractive target for structure-based design of new antiviral agents. PMID- 11927956 TI - Amendments to the farm bill would affect research animal welfare. PMID- 11927957 TI - Reporting ex-post facto pain can be a pain. PMID- 11927958 TI - Enlarged abdomen and ascites in a Syrian hamster: lymphoma. PMID- 11927959 TI - Moving research from the cage to the bedside: the need for IACUC/IRB cooperation. PMID- 11927960 TI - Green fluorescent protein imaging of tumor cells in mice. PMID- 11927961 TI - Physiological and behavioral responses to stress: what does a rat find stressful? PMID- 11927964 TI - One pathologist's reminiscences of the 20th century and random thoughts about the 21st: reflections at the millennium [correction of millenium]. PMID- 11927965 TI - Prevalence of RET/PTC rearrangements in Hashimoto's thyroiditis and papillary thyroid carcinomas. AB - The relationship between Hashimoto's thyroiditis (HT) and follicular cell-derived thyroid cancer remains unclear. Recently, 2 studies reported a 95% prevalence of RET/PTC rearrangements in histologically benign tissue affected by HT, suggesting that multiple occult tumors exist in HT patients with high frequency. We tested the prevalence of RET/PTC rearrangements in 26 HT, in 6 papillary carcinomas arising in the background of HT, and in 27 papillary carcinomas not associated with HT. We detected no RET/PTC rearrangements in HT or papillary carcinomas arising in the background of HT, in contrast to a 33% prevalence among papillary carcinomas not associated with HT. However, the expression of wild-type RET was found in more than half of papillary carcinomas. These results suggest that, if the association between HT and thyroid cancer exists, its molecular basis is different from RET/PTC rearrangement. PMID- 11927967 TI - The association of benign and malignant ovarian adenofibromas with breast cancer and thyroid disorders. AB - An unexpected association with breast cancer and thyroid disorders was found during a review of 91 cases of benign and malignant ovarian adenofibromas. Sixty three tumors were benign, 11 had areas of borderline neoplasms, and 17 had a component of carcinoma. Such tumors were divided into glandular/cystic (61 cases) and papillary (30 cases) according to their gross and microscopic appearance. Among the 61 patients with glandular/cystic adenofibromas, 13 (21%) had breast cancer and 19 (31%) also had thyroid disorders. Among the 30 patients with papillary adenofibromas there were no cases of breast cancer and only 2 patients had thyroid disorders. The average age of the patients with ovarian adenofibroma and breast cancer or thyroid disorders was higher (66 years) than that of patients without breast cancer or thyroid disorders (55 years). More patients with breast cancer and thyroid disorders had bilateral adenofibromas than patients without breast cancer or thyroid disorders. We also reviewed the medical records of 100 patients with ovarian cancer without adenofibroma component, 100 patients with breast cancer, and 100 patients with ovarian and breast cancer. Six percent of patients with ovarian cancer had breast cancer and 16% of each one of these groups had thyroid disorders. This unexpected association found between glandular/cystic adenofibromas, breast cancer, and thyroid disorders might be explained by defects common to these organs. Disorders of some of these organs have been linked by common genetic changes and it is known that these organs are under the influence of similar hormones. Mutations of PTEN have been found in breast and thyroid cancer. The thyroid and ovaries are controlled by glycoprotein hormones of the pituitary gland, which have common alpha subunits. PMID- 11927966 TI - Testicular seminoma: a clinicopathologic and immunohistochemical study of 105 cases with special reference to seminomas with atypical features. AB - In spite of the high curability rates, rare cases of testicular seminoma behave in an unexpectedly aggressive manner. No effective markers are currently available that can predict such uncommon behavior. We studied 105 cases of testicular seminoma on whom the primary resection was performed at the Memorial Sloan-Kettering Cancer Center, New York, to investigate any relationship between morphology, immunohistochemical features, and clinical/pathological stages. Fifty nine percent of the cases presented with pT stage 1 and 74 percent with American Joint Committee on Cancer (AJCC) stage I. In univariate analysis, tumor size, mitotic count, and presence of necrosis showed significant associations with pT stage (p = 0.0001, 0.0001, and 0.04, respectively), and the presence of vascular invasion (p = 0.0001, 0.0001, and 0.02, respectively). In multivariate analysis, both the tumor size and mitotic counts were independent predictors of pT stage (p = 0.0004 and 0.0001) and vascular invasion (p = 0.0004 and 0.0001). When tumors were separated on the basis of architectural and/or cytological atypia into "usual seminomas" and "seminomas with atypia", these were significantly associated with AJCC stage (p = 0.02), and c-kit protein (p = 0.0005) and CD30 expression (p = 0.02). In addition, "seminomas with atypia" also tended to show a higher proliferation activity as judged by Ki67 reactivity (p = 0.06), as well as express the marker of epithelial differentiation, Cam 5.2 (p = 0.09). In summary, we find that the morphologic features in testicular seminomas are associated with factors of clinical relevance. Also, "seminomas with atypia" differ from "usual seminomas" morphologically, present at a higher AJCC stage, and possibly represent an early step in transformation of seminomas toward a more aggressive phenotype. While not proposing a new entity, we suggest that when these atypical features are encountered in an otherwise classical seminoma, investigations must be performed to exclude an early carcinomatous differentiation or even earlier changes toward such a differentiation, such as lack of c-kit protein expression. PMID- 11927968 TI - Renal pelvic carcinoma with unusual appearance simulating amyloidosis (myeloma kidney): a report of five cases. AB - We describe 5 cases of urothelial carcinoma (UC) of the renal pelvis, which grew in a distinctive gross and microscopical pattern into the renal parenchyma. Five patients (2 men and 3 women, mean age 67.4 years) underwent nephrectomy for vague clinical findings. The cut surface of the tumor was white to light gray and the consistency was elastic. The corticomedullary border was indistinct, resulting in an appearance that suggested amyloidosis or myeloma. The renal pelvis showed normal mucosa with areas of dysplastic changes. The tumors spread from the renal pelvis in a diffuse and irregular, infiltrative pattern and surrounded intact glomeruli. Detailed sampling of invasive tumor component showed foci of UC with transitions to clear squamous cells. The predominant clear squamous neoplastic cells had foci of granular eosinophilic cytoplasm and resembled conventional renal cell carcinoma. Four patients were alive and without signs of the disease for 5 months to 4 years after nephrectomy; 1 died of generalized tumor 7 months after nephrectomy. The unusual gross and microscopic features result in varied problems in differential diagnosis, which are discussed herein. PMID- 11927969 TI - The role of thymidine phosphorylase and thrombospondin-1 in angiogenesis and progression of intrahepatic cholangiocarcinoma. AB - Thymidine phosphorylase (TP), an important regulator of angiogenesis, is correlated with progression, metastasis, and prognosis in various types of tumor. In contrast, both positive and negative effects of thrombospondin-1 (TSP-1) on angiogenesis have been reported. In the present study, we examined the expression of TP and TSP-1 in carcinoma cells in 67 primary intrahepatic cholangiocarcinomas (ICCs) immunohistochemically and its correlation with angiogenesis, clinicopathological features, and prognosis. Twenty-six (38.8%) cases were classified as exhibiting positive TP expression. TP expression showed a significant correlation with vascular invasion, lymphatic permeation, perineural invasion, and lymph node metastasis. Thirty-four (50.7%) cases were classified as exhibiting positive TSP-1 expression. TSP-1 expression was significantly correlated with only lymphatic permeation. The microvessel count in positive TP expression cases was significantly higher than that in negative cases. In contrast, the microvessel count in negative TSP-1 expression cases was significantly higher than that in positive cases. Survival in patients who were positive for both TP and TSP-1 expression was significantly poor. Our results suggest that the increased TP expression and decreased TSP-1 expression contribute to angiogenesis, but that the role of angiogenesis in ICC is not closely related to tumor aggressiveness. The TP and TSP-1 expression in ICC may enhance tumor aggressiveness. PMID- 11927970 TI - Images in pathology: low-grade intraductal mammary carcinoma. PMID- 11927971 TI - Images in pathology: oxyuris (enterobius) vermicularis in human cecum and appendix. PMID- 11927972 TI - Multicystic congenital mesoblastic nephroma. AB - This report describes an unusual example of congenital mesoblastic nephroma cellular variant that presented in a 1-week-old neonate as a multicystic tumor of the kidney. Extensive pseudocystic cavitation resulted from progressive accumulation of ground substance in a loosely myxoid tissue composed of stellate- and spindle-shaped cells that compressed and infiltrated renal tissue. The cells of the tumor were positive for vimentin and smooth muscle actin. The patient is alive and well 16 years after surgery. Differential diagnosis from segmental cystic dysplasia, cystic intralobar nephrogenic rest, cystic nephroma, cystic partially differentiated nephroblastoma, cystic nephroblastoma, and cystic clear cell sarcoma of the kidney, all of which may present at this age, is discussed. PMID- 11927973 TI - Endocervicosis of the small intestine. AB - A case of endocervicosis of the small intestine incidentally found as a mass lesion during a gastric bypass surgery is reported. No previous cases of intestinal endocervicosis have been reported in the literature. PMID- 11927974 TI - Plexiform spitz nevus. AB - In 1999, Spatz et al. reported the first 2 cases of a plexiform variant Spitz nevus. We describe another case of this variant on the upper thigh of a 10-year old boy and discuss the differential diagnosis. Histologically, it comprised nodules of epithelioid cells with a plexiform arrangement distributed throughout the superficial and deep dermis. The cells had moderate eosinophilic cytoplasm with vesicular nuclei and prominent nucleoli. Intranuclear inclusions were present focally. The tumor nodules were embedded in a myxoid stroma with intratumoral and peritumoral inflammatory cells including both lymphocytes and eosinophils. There were occasional melanin-containing spindle-shaped cells. An intraepidermal component could not be assessed, since the lesion was ulcerated. The differential diagnosis includes both melanocytic and nonmelanocytic lesions exhibiting a plexiform pattern of growth as well as myxoid lesions. PMID- 11927976 TI - Standardised in vitro electrophysiologic measurements using isolated perfused porcine hearts--assessment of QT interval alterations. AB - During the last decade there have been many clinical case reports on severe arrhythmias following drug administration. These potentially fatal heart rhythm disturbances are often associated with a QT interval prolongation in the ECG. The aim of the present study was to evaluate, whether isolated hemoperfused porcine hearts from slaughter pigs are suitable to detect a drug effect on the ECG. In the absence of test substances heart rate, QT or QRS interval remained fairly constant during 4 hours of perfusion. In contrast, after application of the positive control sotalol a reproducible, concentration-dependent QT interval prolongation as well as adverse effects such as described in clinical case reports were observed. Lidocaine, which served as a negative control, did not change QT interval duration. As a complex model the isolated hemoperfused porcine heart is a versatile system for monitoring drug effects on heart electrophysiology. Therefore, the model might reduce or even replace certain in vivo dog experiments in heart toxicology and safety pharmacology. PMID- 11927975 TI - Dedifferentiated liposarcoma of the pyriform sinus: report of a case and review of the literature. AB - Laryngeal and hypopharyngeal liposarcomas are extraordinarily infrequent tumors. To the best of our knowledge there are fewer than 40 well-documented cases reported to date. Almost all of them are well-differentiated liposarcomas, with only 2 laryngeal-hypopharyngeal dedifferentiated liposarcomas. Dedifferentiated liposarcoma is defined as a well-differentiated liposarcoma with areas of high grade spindle cell nonlipogenic sarcoma. The well-differentiated areas may be of a lipoma-like, sclerosing, or mixed type, and the dedifferentiated areas most frequently are of malignant fibrous hystiocytoma-like type. Despite its commonly pleomorphic histology, dedifferentiated liposarcoma does not behave as aggressively as most pleomorphic sarcomas of adulthood; however, it has the capacity to metastasize, in contrast to its well-differentiated counterpart. We present a case of dedifferentiated liposarcoma arising in the pyriform sinus, an event only twice reported previously in the literature. PMID- 11927977 TI - Multiple-organ harvesting for models of isolated hemoperfused organs of slaughtered pigs. AB - Single- and multiorgan transplantations for the treatment of terminal cardiac, pulmonary, hepatic and renal diseases gain therapeutic importance and transplantation research using mammalian models of isolated perfused organs resembles a major field for the improvement of transplantation techniques. We established a new method of multiple organ harvesting from slaughterhouse pigs which may be used to reduce numbers of laboratory animals. 492 organs (hearts n = 191, lungs n = 108, livers n = 48, kidneys n = 145) were harvested, perfused with autologous blood to prevent artificial perfusion effects and examined for optimal organ harvesting and preservation conditions by analysing organ specific perfusion parameters. For each organ, specific techniques in regard to dissection, cannulation and periods of ischemia have to be applied to guarantee appropriate organ perfusion. In summary, our data indicate that porcine organs obtained by multiple organ harvesting from commercial slaughterhouse animals can be efficiently used for isolated and perfused organ models. The harvesting process did not disturb the commercial use of the animals. Thus, multiple or single organ harvesting may be a useful method to reduce the use of laboratory animals. PMID- 11927978 TI - Novel serum replacement based on bovine ocular fluid: a useful tool for cultivation of different animal cells in vitro. AB - Different mammalian cells in culture have individual nutritional requirements, which are mainly fulfilled by the addition of foetal calf serum (FCS) to the basic medium. Collecting FCS is accompanied with severe animal welfare problems as conscient animals usually are bleeded to death by heart-punctuation without anaesthetics. There exists scientific problems too. Due to the batch-to-batch variability and the relatively high price, different types of serum replacements were introduced. Among them bovine colostrum as a serum substitute for the cultivation of hybridoma cells should be mentioned. The presented experiments were aimed to introduce the simple and effective serum replacement (SR) based on the bovine ocular fluid. Throughout the experiments the bovine ocular fluid alone and in the combination with the sheep's defibrinated plasma and human serum albumin was tested for the growth of different cells growing as a monolayer: (a) Cell lines: WISH (human amniotic cell lines) and VERO. (b) Primary culture: chicken embryonic fibroblasts, human bone-marrow fibroblasts. All growth experiments were performed in parallel with the Foetal Calf Serum (FCS) of three different sources. All types of cells were cultivated in Eagle's medium + antibiotics (Penicillin, Streptomycin, Gentamycin). The most effective was the SR containing approximately 35% of sheep's defibrinated plasma and 1.5% of serum albumin in the bovine ocular fluid. During the experiments 1 and 10% of SR-2.05 or FCS in Eagle's medium were used. After 1, 3 and 6 days of cultivation the cells were counted. The results show that the use of SR-2.05 gives a higher number of cells as compared to most batches of FCS. It is also important that practically no adaptation is needed, meaning that the cells could be grown in Eagle's medium + FCS and in the next passage in Eagle's medium + SR-2.05 and vice versa. PMID- 11927979 TI - [Good cell culture practice--implementation of a relational cell culture database]. AB - The claim for cell culture to provide validable in vitro models for biomedical research postulates evasion of possible fatal record keeping errors. A prototype of a relational computer database for IBM-compatible personal computers using Microsoft(r) Windows 95/98/2000 and NT for administration of cell culture data has been developed using Microsoft(r) Access 98 (Microsoft Corporation, Redmond, USA), -Access Basic, -Visual Basic and Structured Query Language (SQL) (IBM Corporation, Armonk, USA), and was tested successfully. The modular software application manages the many aspects of cell culture laboratory record keeping like detailed information on tissue donor, primary cell isolation/cell line origin, immunohistochemical/molecular biological characterisation, cell countings at passaging/subcultivation/cell aliquotation and cryopreservation. One main feature is a collection of all methods performed at our cell culture laboratory, where linked tables and files store specific informations. Entries into the database are checked via validation rules for correctness to avoid mistakes. The developed prototype has been demonstrated to be an adaptable, reliable tool for improving quality of information storage according to Good Scientific Practice (GSP), Good Cell Culture Practice (GCCP) and general ISO certification trends. PMID- 11927980 TI - Transformation in heterokaryons of Neurospora crassa is nuclear rather than cellular phenomenon. AB - In Neurospora crassa, multinucleate macroconidia are used for genetic transformation. The barrier for such a transformation can be either at the cell membrane level or at the nuclear membrane level. For assessment of these possibilities, a forced heterokaryon (containing two genetically marked nuclei and auxotrophic for histidine) of Neurospora crassa was transformed with a plasmid containing his-3+ gene. The transformants, which could grow without histidine supplementation, were then resolved into component homokaryons to determine into which nucleus or nuclei the plasmid had entered. Our results suggest that the barrier for transformation in Neurospora crassa is at the nuclear level, not at the cell membrane level. In a heterokaryon containing two genetically distinct nuclei, plasmid DNA integrated into only one of the nuclear types at any instance, but never into both nuclear types. Thus, in Neurospora crassa, the competent nucleus is essential for the transformation event to take place, and at a given time only one type of nucleus is competent to take up the exogenous DNA. Genomic Southern analysis showed that the transformants harbor both ectopic and homologous integrations of the plasmid DNA. The type and number of integrations were reflected at the post-translational level, since the specific activity of histidinol dehydrogenase (the translation product of his-3+ gene) was variable among several transformants and always less than the level of the wild type. PMID- 11927981 TI - Sequence polymorphism of dotA and mip alleles mediating invasion and intracellular replication of Legionella pneumophila. AB - Legionella pneumophila inhabit a variety of natural and man-made aquatic environments, where they live primarily as intracellular parasites of protozoans. Given the proper exposure, however, they can cause opportunistic pneumonic infections in humans. The products of two L. pneumophila genes, dotA and mip, are part of the mechanism mediating the initial invasion of eukaryotic cells, and subsequent intracellular survival and multiplication. In this study, DNA polymorphism of the dotA and mip genes was assessed for 17 clinical and environmental isolates by nucleotide sequencing to determine the level of sequence variation, rates of molecular evolution, and history of gene divergence. The mip gene is highly conserved, whereas dotA is extremely variable, with an average level of nucleotide diversity four times greater than that of mip. Gene trees for each locus support a division of the L. pneumophila isolates into two clonal lineages. There are several disagreements between the gene trees suggesting that although L. pneumophila has a clonal population structure, genetic exchange has contributed to genotypic variation among strains in nature. PMID- 11927982 TI - ERIC and REP-PCR banding patterns and sequence analysis of the internal transcribed spacer of rDNA of Stemphylium solani isolates from cotton. AB - The genetic diversity of the Stemphylium solani isolates from cotton was assessed by Enterobacterial Repetitive Intergenic Consensus (ERIC) and Repetitive Extragenic Palindromes (REP)-PCR fingerprinting. Twenty eight monosporic isolates of S. solani from cotton were used along with five isolates from tomato and one isolate of Alternaria macrospora from cotton for comparison. The dendrogram obtained revealed clear differences between the cotton and tomato isolates as well as between the tomato isolates from different geographic regions. The genetic relationships among S. solani isolates were also analyzed by sequencing the internal transcribed spacer (ITS) region of four isolates representing the three ERIC and REP groups. The tomato isolate from the State of Sao Paulo showed a distinct ITS sequence from that of the cotton isolates and tomato isolate from the State of Goias, giving evidence that it belongs to a different genotype of S. solani. This is the first report of the entire sequence of the ITS1-5.8S-ITS2 regions of S. solani. PMID- 11927983 TI - Survival of Enterococcus faecalis in seawater microcosms is limited in the presence of bacterivorous zooflagellates. AB - The survival and persistence of growing and starved cells of Enterococcus faecalis in untreated and differentially filtered (20 microm, 5 microm, 3 microm, 1.2 microm, and 0.1 microm) seawater was analyzed in samples taken at different times over a 1-year period by plate counts and scanning electron microscopy. Whereas seawater filtered through a 0.1-microm mesh was not at all or only slightly bactericidal during incubation at 16 degrees C in the dark, culturability of E. faecalis in the other systems decreased as a function of increasing pore size of the filters. Recovery of culturable, glucose pre-starved cells was always higher than that of cells harvested from the exponential growth phase. Electron microscopic analysis showed that the disappearance of enterococci appeared related to the presence and multiplication of various zooflagellates. PMID- 11927984 TI - Regioselectivity of new bacterial lipases determined by hydrolysis of triolein. AB - The newly identified lipases of 67 bacterial strains, primarily Bacillus and Pseudomonas, from the ARS Culture Collection have been characterized on the basis of their positional specificity for triglycerides (triolein). Lipase was produced by growing the cultures in tryptone-glucose-yeast extract medium for 24 h at 30 degrees C before addition of triglyceride. The lipase was allowed to act on the triglyceride for 3 days before analysis by thin-layer chromatography. Of the bacterial lipases tested, 55 displayed random specificity, 9 were 1,3-specific, and 3 showed no apparent lipase activity under these conditions. PMID- 11927985 TI - Isolation from a shea cake digester of a tannin-tolerant Escherichia coli strain decarboxylating p-hydroxybenzoic and vanillic acids. AB - A facultatively anaerobic, mesophilic, Gram-negative, non-motile, non-sporulated bacterium, designated strain C2, was isolated from an anaerobic digester fed with shea cake rich in tannins and aromatic compounds and previously inoculated with anaerobic sludge from the pit of a slaughterhouse, after enrichment on tannic acid. The straight rods occurred singly or in pairs. Strain C2 fermented numerous carbohydrates (fructose, galactose, glucose, lactose, mannose, maltose, melibiose, raffinose, rhamnose, ribose, saccharose, sorbitol, trehalose, and xylose) and peptides (Biotrypcase, Casamino acids, and yeast extract), producing acid and gas, and had a G + C content of 51.6 +/- 0.1 mol %. Strain C2 was very closely related to Escherichia coli (= DSM 30083(T)) phylogenetically (similarity of 99%), genotypically (DNA homology of 79%), and phenotypically. The isolate tolerated tannic acid (hydrolyzable tannin) and decarboxylated by non-oxidative decarboxylation only p-hydroxybenzoic and vanillic acids to their corresponding phenol and guaicol, under anaerobic and aerobic conditions without further degradation. Adding glucose increased growth and the rate of conversion. High concentrations of p-hydroxybenzoic acid or vanillic acid inhibited growth, and decarboxylation could not occur completely, suggesting phenol toxicity. In contrast, the type strain of E. coli cannot metabolize p-hydroxybenzoic and vanillic acids, anaerobically or aerobically, with or without glucose added. PMID- 11927986 TI - Linezolid is a specific inhibitor of 50S ribosomal subunit formation in Staphylococcus aureus cells. AB - Linezolid is an oxazolidinone compound that has been shown to have impressive antimicrobial activity against a number of Gram-positive bacteria. It inhibits an initiation step of protein synthesis, and its binding site has been shown to be on the 50S ribosomal subunit. Linezolid was tested to see whether would interfere with the formation of the 50S subunit in Staphylococcus aureus cells, since a number of other 50S-specific antibiotics have this second inhibitory function. Linezolid inhibited protein synthesis in S. aureus cells with an IC50 of 0.3 microg/ml. A concentration-dependent decline in cell number with an increase in generation time was found. Pulse-chase labeling studies revealed a specific inhibitory effect on 50S particle formation, with no effect on 30S subunit assembly. The compound inhibited 50S synthesis with an IC50 of 0.6 microg/ ml, indicating an equivalent effect on translation and particle assembly. A postantibiotic effect of 1 h was found when cells were initially treated with the drug at 2 microg/ ml. 50S particle numbers recovered more rapidly than translational capacity, consistent with the increase in viable cell numbers. The inhibitory activities of this novel antimicrobial agent in cells are discussed. PMID- 11927987 TI - Tolerance to cadmium of free-living and associated with marine animals and eelgrass marine gamma-proteobacteria. AB - The tolerance to Cd2+ and possible mechanisms of Cd2+ detoxification by 178 free living bacteria isolated from sea water, associated with marine animals (a mussel Crenomytilus grayanus, a scallop Patinopecten yessoensis), and eelgrass Zostera marina collected in The Sea of Japan and The Sea of Okhotsk have been studied. The concentrations of 25 and 50 mg Cd2+/L were highly toxic and inhibited the growth from 54% to 78% of the total bacteria studied. The free-living bacteria isolated from seawater samples (up to 50%) were tolerant to high concentrations of cadmium. Marine gamma-proteobacteria tolerated Cd2+ by the activation of different detoxifying mechanisms. The strain Halomonas sp. KMM 734 isolated from seawater prevented the uptake of Cd2+ into bacterial cells. The chromosomal cadmium resistance system of Pseudoalteromonas citrea KMM 461 and Marinobacter sp. KMM 181 was found to be similar to class III metallothioneins (also known as phytochelatins). PMID- 11927988 TI - Inhibition of biosynthesis and activity of nitrogenase in Azospirillum brasilense Sp7 under salinity stress. AB - Azospirillum brasilense is a microaerophilic, plant growth-promoting bacterium, whose nitrogenase activity has been shown to be sensitive to salinity stress. Growth of A. brasilense in semi-solid medium showed that diazotrophic growth in N free medium was relatively less sensitive to high NaCl concentrations (200-400 mM) than that in presence of NH4+. Increase in salinity stress to diazotrophic A. brasilense in the semi-solid medium led to the migration of the pellicle to deeper anaerobic zones. Assays of acetylene reduction and nifH- lacZ and nifA- lacZ fusions indicated that salinity stress inhibited nitrogenase biosynthesis more strongly than nitrogenase activity. Under salt stress, the amount of dinitrogenase reductase inactivated by ADP-ribosylation was strongly reduced, indicating that the dinitrogenase reductase ADP ribosyl transferase (DRAT) activity was also inhibited by increased NaCl concentrations. Movement of the pellicle to the anaerobic zone and inhibition of DRAT might be adaptive responses of A. brasilense to salinity stress under diazotrophic conditions. Supplementation of glycine betaine, which alleviates salt stress, partially reversed both responses. PMID- 11927989 TI - Stability of species composition of fecal bifidobacteria in human subjects during fermented milk administration. AB - Bifidobacteria play important roles in human health. However, the influence of exogenous factors on species composition of fecal bifidobacteria is still unclear. The objective of this study was to investigate the effects of fermented milk administration on the species composition of fecal bifidobacteria by molecular biological methods. Fermented milk containing Lactobacillus helveticus was given to seven healthy subjects, and the probiotic effect on human fecal microflora was demonstrated as a significant increase of bifidobacteria and decrease of clostridia by the conventional culture method. Species composition of bifidobacteria in the human fecal microflora was then investigated directly in fecal specimens by the PCR detection method. The species composition of bifidobacteria in the fecal specimens did not change significantly throughout the study period. These findings suggest that the species composition of bifidobacteria remains stable even when fecal microflora is improved by food management. PMID- 11927991 TI - The beta-galactosidase activity in Kluyveromyces marxianus CBS6556 decreases by high concentrations of galactose. AB - In this paper we report on the effect of different concentrations of lactose and galactose in the production of beta-galactosidase by Kluyveromyces marxianus CBS6556. The results clearly demonstrate a decrease in enzyme specific activity during cultivation at high concentrations of L-lactose or D-galactose, despite the fact that these carbohydrates are normally used for induction of the beta galactosidase activity. Therefore, maximum induction of beta-galactosidase in K. marxianus batch cultures was obtained at low concentrations of the inducer carbohydrates, in the range between 0.5 to 15 mM. Those informations can help to design low cost medium with higher beta-galactosidase productivity by K. marxianus cells. PMID- 11927990 TI - Biogenic amine production by Oenococcus oeni. AB - The biogenic amine-producing capability of several Oenococcus oeni strains, originally isolated from different Italian wines, was determined. The amine producing capability was quali-quantitatively variable among the strains: out of the 44 strains investigated under optimal growth conditions, more than 60% were able to produce histamine, at concentrations ranging from 1.0 to 33 mg/L, and about 16% showed the additional capability to form both putrescine and cadaverine, to different extents and variable relative proportions. The amine producing behavior of the strains was confirmed under stress culture conditions, while performing malolactic fermentation. In wine, one randomly chosen strain was very effective in forming putrescine from ornithine. The formation of putrescine from arginine by some strains has been also demonstrated. Consequently, O. oeni can really and significantly contribute to the overall biogenic amine content of wines. Practical consequences of these findings are discussed. PMID- 11927992 TI - The cryosurgical ablation of bone tissue by means of a new miniature cryoprobe -- evaluation of the probe and adaption of the method to in vitro human bone. AB - Until now, modern miniature cryoprobes have been used successfully for the local destruction of soft-tissue tumors without damaging adjacent healthy tissue. In this study, the methodology of cryoablation was applied to bone, and the freezing effect as well as the cooling capacity of the probe were examined. Freezing was performed by cooling one or two probes, with a diameter of 3.2 mm, to -180 degrees C with liquid nitrogen. The cooling capacity of the probes was determined under optic and thermic control in a homogenous reference gel (gelatin), followed by an in vitro measurement on human bone. The simultaneous use of 2 probes resulted in a synergistic effect which produced an almost spherical expansion of frozen area in the homogenous gelatin. In vitro freezing of human tibiae produced equivalent freezing temperatures, with one or two probes, in comparison to the homogenous gelatin. An adequate tissue cooling of bone matrix can be achieved through the use of one or more miniature cryoprobes so that after in vivo testing, the use of this probe could possibly become an alternative or supplement to the surgical resection of pathologic bone processes. PMID- 11927993 TI - Hemicallotasis for medial gonarthrosis: a short-term follow-up of 21 patients. AB - We have prospectively evaluated 21 patients (22 knees; 15 men and 6 women) who underwent hemicallotasis osteotomy (HO), using an external fixator, of the proximal tibia due to medial gonarthrosis. Their median age at the time of operation was 52 (range 39-62) years. The follow- up period was 12-28 months. The Hospital for Special Surgery score (HSS) increased from median 71 preoperatively to 83 points at the follow-up ( p < 0.001). The median hip-knee-ankle angle (HKA) was 187 degrees preoperatively, 177 degrees after distraction, and 180 degrees at the time of follow-up ( p < 0.001). In 5 knees, valgus angulation was not achieved during the distraction phase, and in two-thirds of the knees, there was no valgus at follow-up. The total time in external fixation was median 13 (range 8-16) weeks. Pin tract problems occurred in 12 cases. There was one deep vein thrombosis, and 2 patients had to undergo re-operations due to technical errors. One patient acquired a bacterial arthritis 6 months after the operation. The clinical short-term results with HO seem to be comparable to those reported earlier for closing-wedge osteotomy. However, HO is a trying procedure for the patient as well as for the surgeon, due to frequent minor complications requiring frequent follow-ups. PMID- 11927994 TI - Total hip arthroplasty in patients younger than 30 years of age following developmental dysplasia of hip (DDH) in infancy. AB - Total hip arthroplasty (THA) in young patients is a controversial subject, due to the high failure rates reported in the literature, and even more so in patients with a history of developmental dysplasia of the hip (DDH). A group of 11 patients, all under the age of 30 years at the time of surgery, underwent THA due to congenital dislocation of the hip. Mean age at the time of operation was 23.3 years (range 16-30 years). The mean follow-up period was 9 years (range 3-14 years). The mean preoperative Harris' hip score (HHS) was 56.9 compared with the postoperative HHS of 90.6. Due to aseptic loosening of the cup, 4 patients underwent successful revision arthroplasty. These encouraging medium-term results in our patients suggest that THA may be a good solution for young patients suffering from coxarthrosis due to DDH, at least temporarily, especially when other alternatives, such as arthrodesis or resection arthroplasty, are considered. PMID- 11927995 TI - Hemiarthroplasty or osteosynthesis in cervical hip fractures: matched-pair analysis in 892 patients. AB - Our aim was to compare hemiarthroplasty (HA) and osteosynthesis (OS) in the treatment of cervical hip fractures using matched-pair analysis, especially with regard to different age groups. Data concerning all hip fractures (excluding pathological fractures) at the University Hospitals of Lund in Sweden, where osteosynthesis with LIH hook-pins was used exclusively, and of Oulu in Finland, using mainly cementless Austin-Moore hemiarthroplasty, were registered during 1989-1996 using the same standardized hip fracture forms filled in preoperatively and at 4 months follow-up. Altogether 446 pairs matched for age, sex, place of residence and walking ability at the time of fracture were found. Patients aged 55-80 years seemed to benefit more, with regard to function, from OS than older patients. At 4 months follow-up, 38% of HA and 48% of OS patients lived in their own homes, 16% and 27% were able to walk alone outdoors, and 11% versus 16% were able to walk without any aids, respectively. At 1 year follow-up, mortality was significantly lower among the OS patients, but the reoperation rate was significantly higher. In conclusion, OS is associated with a better function and lower mortality than HA, especially in younger patients, and it is recommended as the primary treatment for cervical hip fractures in patients younger than 80 years and with good ambulatory capacity, whereas the oldest patients can also be safely treated by HA. PMID- 11927997 TI - Hy-Flex II total knee system and range of motion. AB - We developed the Hy-Flex II total knee and ligament balancing system (Hy-Flex II total knee) which provides (1) a flexion angle of at least 120 degrees or more, (2) a range of motion (ROM) comparable to or greater than that before surgery, and (3) occasional full flexion. The system design has several features: small posterior femoral condyle radius, a 4 degrees posterior tilting of the tibial joint surface, and equal tension of the bilateral soft tissues obtained by using a ligament tensor. From September 1997 to June 1999, Hy-Flex II total knee arthroplasty was carried out on 114 joints of 84 patients with rheumatoid arthritis. One year after surgery, the average flexion in our series was 122.1 degrees +/-15.0 degrees. The number of knees operated on which had a flexion angle of 120 degrees or greater was 82 (71.9%) 1 year after surgery among the total of 114 knees. In total, 94 (82.5%) joints obtained the same level or better than the preoperative flexion; 26 (22.8%) attained full flexion 4 weeks after the operation and 15 (13.1%), 1 year after the operation. These results suggest that this system will be able to achieve our aims in almost all rheumatoid knees. PMID- 11927996 TI - Surgical treatment of primary tumors of the sacrum. AB - Twenty-two patients with primary tumors of the sacrum were surgically treated between 1983 and 1997. Seventeen male and 5 female patients were followed up for a mean of 53.6 months (range 12-203 months). The histopathologic diagnoses were giant cell tumor (GCT) in 7 patients, chordoma in 4 patients, aneurysmal bone tumor in 3 patients, chondrosarcoma in 2 patients, osteoblastoma in 2 patients, synovial sarcoma in 2 patients, Ewing's sarcoma in 1 patient, and simple bone cyst in 1 patient. Currettage and thermo- or chemocauterization was applied to 8 patients, a subtotal sacrectomy was done in 11 patients, and total sacrectomy and lumbopelvic stabilization was done in 3 patients. The surgical margins were wide in all patients with GCT. The surgical margins were wide in 3 patients and wide contaminated in 1 patient with chordoma. The 2 patients with chondrosarcoma had high sacral lesions and were managed with total sacrectomy and lumbopelvic fixation. The surgical margin was wide in 1 patient and wide contaminated in the other, who relapsed locally and systemically in the 30th postoperative month. Three patients with aggressive aneurysmal bone cyst and 1 patient with simple bone cyst were managed by curettage and thorough debridement. One patient with low sacral Ewing sarcoma was managed by subtotal sacrectomy with wide margins. The two osteoblastomas were localized to the posterior elements of the sacrum. None of the patients relapsed. Most of the tumors of the sacrum are benign aggressive lesions or low grade malignancies. Intralesional resections in the form of curettage, with the addition of chemo- or thermocauterization, provide a complete cure for benign lesions. In contrast, wide resections are necessary for complete disease control in radio- and chemoresistant malignancies. Nerve root dissection should be performed in order to achieve wide margins. PMID- 11927998 TI - Length of the ribs in patients with idiopathic scoliosis. AB - There have been few reports of actual measurements of rib length in patients with idiopathic scoliosis. We measured the lengths of the 7th to 12th ribs in 28 patients with idiopathic scoliosis with right convex single thoracic curve. The measurements were performed by the multi-projection volume reconstruction method of computed tomography. A significant difference between the left and right side length (laterality) was observed in the 11th and 12th ribs. Laterality was not observed in the 7th, 8th, 9th, and 10th ribs. In this study, it appeared inadequate to conclude that the laterality of rib length caused the scoliosis. Since the 11th and 12th ribs were floating ribs, we speculated that, on the contrary, the scoliosis affected the laterality of the rib length. PMID- 11927999 TI - Corona mortis: incidence and location. AB - The right hemipelvis of 98 patients was examined by angiography to determine the occurrence and location of the corona mortis artery. This arterial anastomosis was found in 28.5% (28 of 98). Its incidence was 30.5% (18 of 59) in men and 25.6% (10 of 39) in women; this difference was not significant ( p>0.05). The distance from the symphysis pubis to the anastomotic artery averaged 33.4 mm (range 21.4-41 mm). It was 31.8 mm (range 21.4-39.3 mm) in men and 36.2 mm (range 25-41 mm) in women; this difference was significant ( p<0.05). PMID- 11928000 TI - Conservative functional treatment of ankle fractures. AB - Thirty-eight patients (mean age 49 years; range 19-91 years; nine of them over 60 years; 28 women, 10 men) suffering from an isolated Weber B fracture with a dislocation of less than 1 mm underwent functional therapy using a pneumatic ankle brace and were included in a prospective study. The clinical outcome was measured according to the Olerud-Molander ankle score. Functional therapy was finished in 34 cases successfully. Twenty-one patients were scored after 17 months on average (range 8-27 months) with the Olerud-Molander ankle score. A very good result was seen in 18 patients, including 12 with 100 points, a complete remission. The remaining 3 patients showed good results (1 had 90, 2 had 85). However, functional treatment failed in 4 cases due to secondary dislocation. These patients underwent surgery without further complications. The control group, 31 operated patients, did not show as good results. Functional therapy of stable Weber-B ankle fractures appears to be superior to surgery. We were able to avoid surgery in 90% of our patients and got better results than with patients undergoing open reduction and internal fixation. PMID- 11928001 TI - Treatment of focal articular cartilage lesions of the knee with autogenous osteochondral graftsA 2- to 4-year follow-up study. AB - A retrospective study of 15 patients with 16 knees who underwent osteochondral autografts for focal full thickness articular cartilage defects of the knee with 2- to 4-year follow-up showed 80% good or excellent clinical results. There was no correlation of the clinical results with the underlying diagnoses, including osteonecrosis, osteochondritis dessicans and traumatic cartilage defect, or a size of the lesion smaller than 600 mm(2). However, cartilage lesions larger than 600 mm(2) were associated with increasing fibrous tissue formation and fissuring between the grafts and the host tissues and poor results. The improvement in symptoms appeared time-dependent, ranging from 6 to 16 weeks, suggesting that postoperative protection of the graft is warranted. There was no radiographic progression of degenerative changes of the knee on the medium-term follow-up. Therefore, an autogenous osteochondral graft is considered a good method in the treatment of knees with moderately sized articular cartilage defects. PMID- 11928002 TI - Resection replantation of the upper limb for aggressive malignant tumors. AB - Stage IIB malignant tumors of the upper limb have been traditionally treated by amputation or disarticulation. There have been isolated reports on the technique of segmental resection of the tumor-bearing segment complete with the skin, and replanting the distal arm or forearm with or without neurovascular repair. The present paper describes four cases in which a wide resection margin was achieved in all by resecting the affected cylinder of the limb. Functional reconstruction was performed by appropriate tendon transfer. The main vessels and nerves were dealt with according to the findings revealed by preoperative investigations. If they had to be sacrificed, end-to-end suture was performed, but if the main nerves could be spared, it greatly enhanced the functional outcome. Local and systemic recurrences occurred in one case, and systemic recurrence occurred in another case. The other two cases remained disease-free at more than 4 years' follow-up. This operation is as radical as amputation, while the esthetic and functional results are equivalent to those of resection-arthrodesis. PMID- 11928003 TI - Acquired pelvic anomaly preventing sexual intercourse in a female patient. AB - This report describes a patient with dyspareunia related to an acquired pelvic anomaly that was treated surgically. PMID- 11928005 TI - Avulsion fracture of the flexor digitorum profundus tendon ('Jersey finger') type III. AB - Avulsion fracture of the flexor digitorum profundus tendon (Jersey finger) is an uncommon injury. This injury involves mainly young and active patients, especially during sports activities. It is classified into three types according to the extent of the tendon retraction. Type III Jersey finger is very rare, and only a few cases have been described so far.In this article, we report a case of a type III Jersey finger. Rapid diagnosis accompanied by surgery which included open reduction and internal fixation with 2 mini screws, followed by physiotherapy, led to restoration of a full range of motion of the involved digit. PMID- 11928004 TI - Isolated ulnar radiocarpal dislocation. AB - Isolated radiocarpal dislocation is quite rare. A patient with an isolated ulnar radiocarpal dislocation was treated with closed reduction and external fixator. The patient was followed-up for 20 months. The functional results were successful,but a secondary ulnar translation of the corpus developed. PMID- 11928006 TI - Isolated type IIIA fracture of the coronoid process of ulna. A case report and brief review of literature. AB - A very rare, Regan and Morrey's type IIIA fracture of the coronoid process of ulna is presented and discussed with a review of the literature. The operative approach to be used is emphasised. Open reduction and internal fixation render the elbow stable, and a good range of motion can be achieved if such fractures are fixed and mobilised early. PMID- 11928007 TI - Revision hip arthroplasty in a patient with a hip rotationplasty (type B III b). AB - We report for the first time a patient who underwent a revision hip arthroplasty after a hip rotationplasty (Winkelmann type B III b) because of a chondrosarcoma in 1988. The clinical symptoms and radiographic findings (X-ray, bone scintigraphy, arthrography) of the aseptic loosening of the acetabular component are presented in detail. Furthermore, we present the functional results after hip revision. PMID- 11928008 TI - Operative treatment in case of a closed rupture of the anterior tibial tendon. AB - Closed rupture of the tibial anterior tendon is a rare clinical entity. Case reports in the literature reveal a total of only 49 cases up to the year 2000. According to these reports, the age group affected is 50 to 70 years old, and there are more men than women affected. Although the functional limitation is quite considerable, late diagnosis is common. An appropriate clinical examination, including an exact history taking, should lead to the right diagnosis. Ultrasound examination and magnetic resonance imaging (MRI) may be helpful. 'Restitutio ad integrum' can only be achieved by operative treatment. If technically possible, reinsertion of the tendon directly into bone or direct tendon repair is preferred. After delayed diagnosis, a secondary reconstruction through tendon transfer or transplantation is often necessary. A 64-year-old woman presented with pain and swelling in the area of the ankle joint 5 months after falling. She showed insecurity in walking, and the heel-walk could not be demonstrated. The distal neurovascular function was intact. The area of the retinaculum showed a swelling, and the tendon was not palpable in comparison with the other forefoot. An intact tendon could not be seen by ultrasound, and MRI confirmed these findings. A complete rupture was noted during the operative revision. The proximal and the distal tendon stumps were found to be thickened and knotted, the proximal stump was also atrophic. An augmented tenoplasty was performed. Afterwards, the tendon was tense in the neutral position. The lower leg was put in a plaster cast for 6 weeks, followed by physiotherapy. Ten months after the operation, the tendon was palpable in the correct position, the dorsal extension was powerful, and the patient did not experience any difficulty. Rupture of the anterior tibial tendon is a rare clinical entity and should be considered in the differential diagnosis of pain in the area of the ankle joint. An early operative treatment is advantageous. PMID- 11928012 TI - Is laparoscopic refundoplication feasible in patients with failed primary open antireflux surgery? AB - BACKGROUND: Due to the widespread availability and acceptance of minimal-access surgery, laparoscopic antireflux surgery has become the standard procedure for the treatment of severe gastroesophageal reflux disease (GERD). However, open and laparoscopic antireflux procedures sometimes result in failure, so that redosurgery is required in some cases. The aim of this prospective study was to evaluate the surgical outcome and quality of life of patients who underwent refundoplication after the failure of primary open antireflux surgery. METHODS: Twenty patients with a mean age of 52 years (range, 33-69) underwent laparoscopic refundoplication after primary open antireflux surgery. Four of them had undergone surgery twice previously. Preoperative and postoperative data, including esophageal manometry, 24-h pH monitoring, and assessment of quality of life, were reviewed prospectively. Quality of life was evaluated using the Gastrointestinal Quality of Life Index (GIQLI). RESULTS: In 18 patients (90%), the reoperation was completed successfully laparoscopically. Two others (10%) required conversion to an open procedure. One of them had an injury of the gastric wall; in the other case, severe bleeding of the spleen necessitated the conversion. The average operating time was 245 min. Preoperatively, the main symptoms were recurrent reflux in 14 cases and a combination of re-reflux and dysphagia in six cases. The anatomic findings were telescope phenomenon (n = 6), hiatal disruption (n = 10), and wrap breakdown (n = 4). Postoperatively, two patients suffered from dysphagia and required pneumatic dilatation. The lower esophageal sphincter (LES) pressure increased significantly from a preoperative value of 6.08 mmHg to 12.2 mmHg at 3 months and 11.9 mmHg at 1 year after surgery. The DeMeester score decreased from a preoperative value of 69.8 to 17.1 at 3 months and 14.6 at 1 year postoperatively. The GIQLI score increased from a preoperative value of 84.9 points to 119.6 points at 3 months and 120.1 points at 1 year. CONCLUSION: Laparoscopic refundoplication after the failure of a primary open intervention is an effective procedure that can be performed safely by experienced laparoscopic surgeon. The procedure yields excellent functional results and leads to significant improvement in the patient's quality of life. PMID- 11928013 TI - Endoscopic augmentation of the cardia with a biocompatible injectable polymer (Enteryx) in a porcine model. AB - BACKGROUND: Endoscopic approaches to restore the gastroesophageal barrier in patients with gastroesophageal reflux disease (GERD) are presently undergoing clinical trial. The aim of the study was to demonstrate the feasibility, durability, safety, and antireflux efficacy following augmentation of the cardia with a biocompatible injectable polymer (Enteryx). METHODS: Augmentation was performed in 12 Yucatan mini-pigs. The cardia was injected circumferentially with 1-1.5 ml of Enteryx at three or four sites. Four groups of three animals each were killed at 2, 6, 12, and 24 weeks following augmentation. Gastrointestinal endoscopy and esophageal manometry were performed preoperatively and postoperatively. Competency was determined as the intragastric pressure (yield pressure) and volume (yield volume) needed during gastric distension with air and water to result in equalization of gastric and esophageal pressure. Comparisons were made with a group of noninjected animals (n = 6). RESULTS: All animals had a normal eating pattern; none showed any evidence of vomiting or regurgitation. The median injection volume was 4 ml (range, 1-8). At autopsy, implants were found in 83% of the animals. Intramuscular placement of the implant was durable, whereas sloughing occurred if the implant was placed submucosally. The mechanical properties of sphincter length and pressure were unaffected by the injection. The median yield pressure of the animals that survived for >6 weeks (21.4 mmHg) was significantly greater (p = 0.049) than the animals that survived for <6 weeks (4.5 mmHg) and greater (p = 0.054) than the control animals (9.1 mmHg), suggesting that the healing process was associated with reduced distensibility of the cardia. CONCLUSIONS: Augmentation of the cardia with an injectable polymer (Enteryx) is simple, safe, and durable. Early studies suggest that alteration in the distensibility and geometry of the gastroesophageal junction may provide antireflux protection. PMID- 11928014 TI - Revisional surgery after failed laparoscopic anterior fundoplication. AB - BACKGROUND: Partial fundoplication may have functional advantages over a circumferential wrap but the reconstruction is more complex. Revisional surgery for recurrent reflux may be more difficult because of the additional suturing involved in the original operation. We report experience with revisional surgery in a large cohort of patients who had undergone laparoscopic anterior fundoplication and hiatal repair. METHODS: Between August 1993 and September 1999, 11 (3.5%) of 309 patients who had laparoscopic anterior fundoplication for uncomplicated gastroesophageal reflux disease required revisional surgery (1 open and 10 laparoscopic revisions). Data were retrieved from a prospective database supplemented by a postal questionnaire following the second operation. RESULTS: The operative findings were posterior hiatal disruption (n = 9), anterior paraesophageal hernia (n = 1), and inadequate initial esophageal mobilization (n = 1). There were no conversions to open surgery in the laparoscopic group. Ten (91%) of the respondents described the outcome of their repeat procedure as either good or excellent. All patients would recommend the repeat procedure to patients with similar symptoms. CONCLUSIONS: Revisional surgery after laparoscopic anterior fundoplication can be performed safely with a good outcome. Modifications to technique both in the primary procedure and for revision may decrease the incidence of early technical failure. PMID- 11928015 TI - The significance of pH and manometric testing after laparoscopic fundoplication. AB - Laparoscopic antireflux surgery has become the standard operation for gastroesophageal reflux disease (GERD). This study examined the outcomes of laparoscopic antireflux surgery, hypothesizing that both subjective symptoms and objective pH would correlate with manometric parameters to reflect the absence of reflux after fundoplication. We evaluated 56 patients who underwent laparoscopic antireflux surgery. Preoperative and postoperative symptoms were documented by chart reviews and confirmed by telephone interviews with the patient. Preoperative pH probe and esophageal manometry studies were compared with postoperative studies performed 3 to 6 months after fundoplication. Subjective symptoms were correlated with objective measurements of acid reflux and lower esophageal sphincter pressure (LESP). The follow-up period was 3 to 29 months. Symptomatic improvement was seen in 91% of patients, and good to excellent improvement in preoperative symptoms was cited. Postoperatively, there was significant improvement in percentages of upright supine times when esophageal pH was less than 4 (p <0.001). There was an increase in LESP from an average of 16.9 mmHg preoperatively to 22.7 mmHg postoperatively (p <0.001). There was no correlation between postoperative LESP and symptoms or LESP and 24-h pH results. However, there was a predictive correlation between LESP and postoperative heartburn symptoms (p <0.001). These findings imply that symptom follow-up evaluation is adequate in the asymptomatic patient after laparoscopic fundoplication, and that routine physiologic testing is not necessary. PMID- 11928016 TI - Comparison of orthodox versus off-optical axis endoscopic manipulations. AB - BACKGROUND: During complex laparoscopic operations, the surgeon often has to use both instruments to one side of the telescope (off-optical axis work). This experimental study was undertaken to compare this orthodox versus the off-optical axis endoscopic manipulations regarding the performance parameters and motion analysis and muscle work of the surgeon's dominant upper limb. METHODS: Ten surgeons participated in the study; each sutured 50-mm enterotomy in pig's small bowel in each of three setups: (1) in-optical axis manipulation (one instrument on either side of the laparoscope) (2) off-optical axis manipulation (both instruments on one side of the laparoscope to the dominant hand of the surgeon), and (3) off-optical axis manipulation both instruments on one side of the laparoscope on the nondominant side). The main outcome measures were the placement error score, execution time, leakage pressure, motion analysis, and telemetric electromyography parameters of the surgeon's dominant upper limb. RESULTS: There was no significant difference in all parameters of performance, muscle work, and fatigue between setup 1 and setup 3. However, marked degradation of all parameters of performance with increased muscle work and fatigue was observed with setup 2 compared to setups 1 and 3. The reason for the deterioration with setup 2 is related to the altered "monitor display angle" which are different from the actual physical angles. With this setup, the instrument-to-target physical angle of 30 degrees appears on the screen as ?30 degrees and this disturbs both the manipulation and the azimuth angles obscuring the needle-tissue entry point. In addition, the instrument casts a shadow on its medial side and this tends to obscure the exact relations between instrument, needlepoint, and the tissue. CONCLUSIONS: Off-optical axis work is a good alternative to the orthodox setup provided the instruments are placed to the nondominant hand. The marked degradation in performance encountered during off optical axis work to the dominant hand of the surgeon is due to the resulting altered monitor display angles. The importance of these monitor display angles in influencing task performance has been previously overlooked. PMID- 11928017 TI - Comparison of video trainer and virtual reality training systems on acquisition of laparoscopic skills. AB - Training on a video trainer or computer-based minimally invasive surgery trainer leads to improved benchtop laparoscopic skill. Recently, improved operative performance from practice on a video trainer was reported. The purpose of this study was three fold: (a) to compare psychomotor skill improvement after training on a virtual reality (VR) system with that after training on a video-trainer, (VT) (b) to evaluate whether skills learned on the one training system are transferable to the other, and (c) to evaluate whether VR or VT training improves operative performance. For the study, 50 junior surgery residents completed baseline skill testing on both the VR and VT systems. These subjects then were randomized to either a VR or VT structured training group. After practice, the subjects were tested again on their VR and VT skills. To assess the effect of practice on operative performance, all second-year residents (n = 19) were evaluated on their operative performance during a laparoscopic cholecystectomy before and after skill training. Data are expressed as percentage of improvement in mean score/time. Analysis was performed by Student's paired t-test. The VR training group showed improvement of 54% on the VR posttest, as compared with 55% improvement by the VT group. The VR training group improved more on the VT posttest tasks (36%) than the VT training group improved on the VR posttest tasks (17%) (p <0.05). Operative performance improved only in the VR training group (p <0.05). Psychomotor skills improve after training on both VR and VT, and skills may be transferable. Furthermore, training on a minimally invasive surgery trainer, virtual reality system may improve operative performance during laparoscopic cholecystectomy. PMID- 11928018 TI - Efficiency of manual versus robotical (Zeus) assisted laparoscopic surgery in the performance of standardized tasks. AB - BACKGROUND: The objective of this study was to compare the efficiency of manual and robotically assisted laparoscopic surgery. METHODS: To evaluate the surgical efficiency in a set of basic endoscopic movements, 20 medical students without any surgical experience were selected to perform at random a set of laparoscopic tasks either manually or robotic assisted (Zeus). This task consisted of dropping beads into receptacles, running a 25-cm rope, capping a hypodermic needle, suturing, and performing a laparoscopic cholecystectomy on a cadaver liver of a pig. A quantitative time-action analysis was performed to evaluate the efficacy and skill performance in terms of time and the number of actions. RESULTS: The dropping beads exercise and the laparoscopic cholecystectomy required more time when performed with robotic assistance, as compared with manual performance (respectively, median, 78.5 s; range, 63 - 122 s vs median, 144.5 s; range, 100 - 169 s; p <0.01 and median, 34.0 min; range 11-44 min vs median, 46.5 s; range, 21 - 79 min; p = 0.05). A tendency toward fewer total actions in all the robotically assisted exercises was observed. However, significance was shown only in the rope passing task (median, 71; range, 59 - 87 vs median, 62; range, 57-80; p = 0.05). Grasping the beads, the rope, and either the needle or the cap were tasks that required fewer actions to complete when performed with robotically assistance (respectively, median, 11; range, 10 - 14 vs median, 12.5; range, 11 - 15; p <0.01; median, 56; range, 55 - 60 vs median, 60.5 min; range, 55 - 65; p = 0.03, and median, 6; range, 4 - 21 vs median, 10.5; range, 6 - 38; p = 0.02). As compared with the robotically assisted rope-passing exercise, more failures were made in the manually performed procedure (p = 0.03), mainly caused by unintentional dropping of the rope (p = 0.02). CONCLUSIONS: Robotically assisted laparoscopic surgery by participants without any surgical experience might require more time, but actions can be performed equally or more precisely as compared with manual laparoscopic surgery. PMID- 11928019 TI - An ergonomic study of the optimum operating table height for laparoscopic surgery. AB - BACKGROUND: Laparoscopic surgery requires the use of longer instruments than open surgery, thus changing the relation between the height of the surgeon's hands and the desirable height of the operating room table. The optimum height of the operating room table for laparoscopic surgery is investigated in this study. METHODS: Twenty-one surgeons performed a two-handed, one-fourth circle cutting task using a laparoscopic video system and laparoscopic instruments positioned at five instrument handle heights relative to subjects' elbow height (-20, -10, 0, +10, and +20 cm) by adjusting the height of the trainer box. Subjects rated the difficulty and discomfort experienced during each task on a visual analog scale. Skin conductance (SC) was measured in Micromhos via paired surface electrodes placed near the ulnar edge of the palm of the right (cutting) hand. The mean electromyographic (EMG) signal from the right deltoid and trapezius muscles was measured. Arm orientation was measured in three dimensions using a magnetometer/accelerometer. Signals were acquired using analog circuitry and digitally sampled using a National Instruments DAQCard 700 connected to a Macintosh PowerBook 5300c running LabVIEW software. Statistical analysis was carried out by analysis of variance and post hoc testing. RESULTS: Statistically significant changes were found in the subjective rating of discomfort (p <0.002), deltoid EMG (p <0.0006), trapezius EMG (p <0.0001), and arm elevation (p <0.0001) between instrument handle heights. SC values and task times did not change significantly. Discomfort and difficulty ratings were lowest when instrument handles were positioned at elbow height. EMG values and arm elevation all decreased with lower instrument height. CONCLUSION: This study suggests that the optimum table height for laparoscopic surgery should position the laparoscopic instrument handles close to surgeons' elbow level to minimize discomfort and upper arm and shoulder muscle work. This corresponds to an approximate table height of 64 to 77 cm above floor level. A redesign of current operating room tables may be required to meet these ergonomic guidelines. PMID- 11928020 TI - Hand-assisted laparoscopic surgery (HALS) for live donor nephrectomy is more time and cost-effective than standard laparoscopic nephrectomy. AB - BACKGROUND: Hand-assisted laparoscopy (HALS) was introduced to increase the safety of living donor nephrectomies. Herein we evaluate the first HALS living donor nephrectomies performed at our center. METHODS: Traditional laparoscopic nephrectomies (TLS) (n = 11) and HALS nephrectomies (n = 11) were included in the study. One patient from the TLS group was excluded because the operation was converted to open nephrectomy. We compared the operating times (OT) and warm ischemia times (WIT) for the two procedures and calculated the operating costs. RESULTS: Mean OT was 270 min in the TLS group and 197 min in the HALS group; thus, there was, a significant reduction of 27% with HALS. WIT was 297 sec for the TLS group and 214 sec for the HALS group, for a reduction of 28%. Costs were also lowered for HALS. CONCLUSION: In addition to shortening both OT and WIT, HALS enhances the safety margin of the procedure, especially during trocar placement. It is further helpful in preventing torsion of the kidney and controlling potential bleedings, as well as during vascular stapling and kidney removal. PMID- 11928021 TI - Hand-assisted laparoscopic splenectomy (HALS) in cases of splenomegaly: a comparison analysis with conventional laparoscopic splenectomy. AB - BACKGROUND: Laparoscopic splenectomy (LS) is considerably more difficult to perform when the spleen is enlarged. The new technique of hand-assisted designed technique aimed to assist laparoscopic surgery allows the surgeon to insert his or her hand into the abdomen while maintaining the pneumoperitoneum, thus recovering the tactile sensation lost in conventional laparoscopic surgery. OBJECT: In this study, we compared the immediate results of conventional LS and hand-assisted LS (HALS) in cases of splenomegaly. METHODS: Between February 1993 and August 2001, 200 LS were attempted at two university hospitals. In 56 cases, splenomegaly (final spleen weight >700 g) was observed clinically or detected on radiological examination. We compared the first 36 patients operated on by conventional LS (group I) with the last consecutive 20 patients, who underwent HALS (group II). The study parameters were operative time, conversion rate, transfusion rate, morbidity and length of hospital stay. RESULTS: The groups were comparable in terms of age (58 +/- 13 [ranges, l9-82] vs 58 +/- 16 years [range, 44-84] (ns), diagnosis, and spleen weight (1425 +/- 884 [range, 700-3400]) vs 1753 +/- 1124 g [range, 720-4500] (ns). HALS was associated with less morbidity (36% vs 10%) (ns), a shorter operative time (177 +/- 52 [range, 95-300]) vs 135 +/- 53 min [range, 85-270] (p <0.009), and a shorter hospital stay (6.3 +/- 3.3 [range, 3-14]) vs 4 +/- 1.2 [range, 2-7] days (p <0.05). CONCLUSION: In cases of splenomegaly, HALS assisted laparoscopic surgery significantly facilitates the surgical maneuvers during LS while maintaining the advantages of a purely laparoscopic approach. PMID- 11928022 TI - Hand-assisted laparoscopic low anterior resection: initial experience with a new procedure. AB - BACKGROUND: Laparoscopic low anterior resection for rectal cancer has never gained wide acceptance among general surgeons, mainly due to the technical difficulties encountered during pelvic dissection. It has therefore been stated that these patients should undergo open rather than laparoscopic surgery. Hand assisted laparoscopic surgery (HALS) is a new technique that has the potential to overcome many of the existing limitations of pure laparoscopy. In the treatment of rectal cancer, HALS could reproduce an operative setting similar to that of the open approach. METHODS: To assess the technical feasibility of hand-assisted laparoscopic low anterior resection for rectal cancer and evaluate potential benefits and drawbacks of this new procedure, a pilot study was conducted at a university hospital on 16 consecutive patients during a 12-month period. Only patients with extraperitoneal rectal cancer were included in this series. Patients' clinical data, operative time, conversion rate, complications, and early outcome measures were prospectively examined. RESULTS: There were 9 men and 7 women. The average +/- SD operation time was 238 +/- 38 min. Conversion to open surgery was never required. Ten of 16 patients were off pain medication on the third postoperative day. Eight were able to walk the day after surgery. Three minor postoperative complications were recorded. Mean postoperative stay for patients without complications was 5.6 +/- 1.4 days. CONCLUSION: From a technical standpoint, the reported hand-assisted procedure makes pelvic dissection during laparoscopic low anterior resection almost equivalent to the laparotomic operation. The incision for hand access that is needed with this technique does not seem to compromise the quick recovery of patients undergoing purely laparoscopic procedures. PMID- 11928023 TI - The changed histologic paradigm of colorectal polyps. AB - BACKGROUND: Previous literature has recorded the preponderance of hyperplastic over neoplastic polyps. This study evaluated the histopathologic characteristics of colonic polyps, excised during colonoscopic polypectomy, and further determined their relationship to age, location, and gender. METHODS: Of 5132 colonoscopies reviewed between 1976 and 1999, 757 were performed on 582 patients who had polyp removal. Patients with previous colon resection or incomplete cecal intubation were excluded. RESULTS: The mean age was 67 +/- 11 years for men and 66 +/- 11 years for women. Of the 1050 lesions histologically analyzed, 871 (83.0%) were neoplastic, 129 (12.3%) were hyperplastic, and 50 (4.8%) were miscellaneous lesions (29 inflammatory polyps, 14 lipomas, 2 leiomyomas, 1 juvenile polyp, and 4 no pathology identified). Hyperplastic polyps were always less than 1 cm (with one exception) and were located predominantly in the left colon, the majority residing in the sigmoid colon. Peak prevalence of hyperplastic polyps occurred in the 50-70 years old age group. Of the neoplastic polyps, 566 (65.0%) were tubular, 225 (25.8%) villotubular, 63 (7.2%) villous adenomas, 4 (0.5%) mixed adenomatous hyperplastic polyps, and 12 (1.4%) invasive carcinomas. The peak prevalence of neoplastic polyps occurred in the same age group as did hyperplastic polyps. Even though adenomatous polyps outnumbered hyperplastic polyps throughout the colon and within each age group, a greater percentage of hyperplastic polyps were found distally and in younger patients compared to location and age groups for neoplastic polyps. CONCLUSION: Adenomatous polyps outnumber hyperplastic polyps 7:1, even in the distal colon. Even small polyps seen during colonoscopy should be removed and subjected to histologic analysis because of the advisability of follow-up examinations of patients with neoplastic polyps. The increase in the incidence of neoplastic polyps beginning at the age of 50 years supports the need for colonoscopy in these individuals. PMID- 11928024 TI - Oncologic implications of laparoscopic and open surgery. AB - Although instrumental manipulation and mechanical tumor cell spillage seem to play the major role in port-site metastases from laparoscopic cancer surgery, minimally invasive procedures are used more and more in the resection of malignancies. However, port-site metastases also have been reported after resection of colon cancer in International Union Against Cancer (UICC) stage I [2, 14]. Therefore, changes in the peritoneal environment during laparoscopy also might influence intra- and extraperitoneal tumor growth during laparoscopy and pneumoperitoneum. Different results of experimental studies presented at the Third International Conference for Laparoscopic Surgery are analyzed and discussed. PMID- 11928025 TI - Advanced proximal colon cancer. AB - BACKGROUND: Two recent studies have documented that sigmoidoscopy as a screening tool for colorectal cancers may miss advanced proximal colonic neoplasms. The purpose of this study was to assess the prevalence of distal synchronous lesions in patients with proximal colon cancer. We sought to determine if screening sigmoidoscopy would have missed these proximal colon cancers. METHODS: Data were collected on all patients (n = 305) diagnosed with colorectal cancer over a 6 year period. Patients were stratified by age, sex, tumor location, presenting complaint, AJCC stage, and TNM classification. The colonoscopy results of patients diagnosed with proximal colon cancer were analyzed to determine the incidence of synchronous distal colon lesions. RESULTS: Proximal colon cancer was diagnosed in 88 patients (29%). Of those studied, 45 (54%) did not have synchronous distal lesions detected by colonoscopy. The patients with proximal colon cancer were elderly (mean age 67), had advanced tumor size [59 patients (67%) T3/T4], and had advanced AJCC stages [37 patients (42%) stage III/IV]. Nearly all patients [84 (95%)] with proximal colon cancer were symptomatic. CONCLUSION: In this study, the majority of patients with proximal colon cancer did not have a synchronous lesion in the distal colon. Current screening methods for colon cancer based on sigmoidoscopy would not have identified these proximal lesions. These findings support the incorporation of screening colonoscopy in protocols designed to identify early colon cancer. PMID- 11928026 TI - Risk factors for gallstone disease in the laparoscopic era. AB - BACKGROUND: The risk factors for gallstone disease are well known, but they have not been updated to take the development of better ultrasound technology and the advent of laparoscopic surgery into consideration. METHODS: We compared two groups of patients who underwent ultrasound-one group (n = 100) who underwent cholecystectomy after ultrasound revealed the presence of gallstones and a control group (n = 107) in whom no gallstones were shown on ultrasound. RESULTS: Seven patients in the control group refused to participate in the study; otherwise, the groups are sequential. Age in the surgery group was 51 years (+/- 16) vs 50 (+/- 16) for the control group. The percentage of female patients was 59% and 52%, respectively (p = ns). Body mass index was 32 (+/- 8) and 28 (+/- 6), respectively (p = 0.013). Parity > 2 was 0.49% and 0.37%, respectively (p = 0.000001). The number who breast-fed at least one child was 17 (24%) and eight (12%), respectively (p = 0.03). Oral contraceptive use was 37 (52%) and 17 (22%), respectively (p = 0.0005). Primary relatives who had had gallbladder surgery was 0.68 (+/- 1) and 0.35 (+/- 0.6), respectively (p = 0.02). CONCLUSION: Body mass index, breast-feeding, oral contraceptives, parity > 2, and family history were found to be risk factors for gallstone disease. Age and female sex were not, probably due to selection bias. PMID- 11928027 TI - Laparoscopic cholecystectomy. AB - BACKGROUND: This study was undertaken to determine if patients undergoing laparoscopic cholecystectomy may be discharged home 4 h postoperatively with similar outcomes as patients admitted overnight. METHODS: Patients were randomized to an outpatient group (OP), consisting of patients who were discharged after a 4-h stay in the Post Anesthesia Care Unit (PACU), or to an inpatient group. Variables compared between the two groups included patient demographics; degree of postoperative pain, nausea, vomiting, and patient satisfaction; amount of pain and nausea medication taken; and number of phone calls, readmissions, or complications. Statistical analysis was performed with students t-test, Fisher's exact test, and Wilcoxon's signed rank and rank sums tests as appropriate. RESULTS: Eighty patients were initially enrolled. Two were converted and 4 required admission after being randomized to the OP group. Patients in the OP group received more oral pain medication prior to PACU discharge. Degree of pain, number of phone calls, readmission and complication rates, and patient satisfaction were similar between both groups. Of the 4 unexpected admissions, all were identified within the 4-h PACU stay. CONCLUSIONS: Patients undergoing laparoscopic cholecystectomy who are discharged home 4 h postoperatively will experience the same satisfaction with no increase in complications as patients admitted overnight. PMID- 11928028 TI - Microlaparoscopic vs conventional laparoscopic cholecystectomy: a prospective randomized double-blind trial. AB - BACKGROUND: Downsizing the port incisions may reduce pain after laparoscopic cholecystectomy. METHODS: In a double-blind controlled study, 60 patients were randomized to undergo either microlaparoscopic cholecystectomy using one 10-mm and three 3.5-mm trocars (3.5-mm LC) or traditional laparoscopic cholecystectomy using two 10-mm and two 5-mm trocars (LC). Incisional pain at each port incision and overall pain were recorded for 1 week after the operation. Fatigue, nausea and vomiting, pulmonary function, and cosmetic results were also measured. RESULTS: Data from 52 patients were analyzed; eight patients were excluded from the study for various reasons. One patient was converted from 3.5-mm LC to LC due to technical problems with the 3.5-mm optic. In the 3.5-mm LC group (n = 25), incisional pain was significantly decreased in the 1st postoperative week as compared with the LC group (n = 27) (p <0.01). In both groups, pain scores at the supraumbilical 10-mm port were significantly higher compared with other port sites (p <0.05). The cosmetic results were significantly better in the 3.5-mm LC group (p <0.01). There were no significant differences in any of the other variables. CONCLUSION: The use of 3.5-mm trocars is feasible in LC, and it both reduces incisional pain and improves the cosmetic result. PMID- 11928029 TI - Selective operative cholangiography and Perioperative endoscopic retrograde cholangiopancreatography (ERCP) during laparoscopic cholecystectomy: a viable option for choledocholithiasis. AB - BACKGROUND: Most common bile duct (CBD) stones can now be removed by minimally invasive methods using pre- or postoperative endoscopic sphincterotomy. A few centers explore the bile duct laparoscopically, but not every hospital has access to the equipment for magnetic resonance (MR) cholangiopancreatography to diagnose ductal stones. This study shows the results of an alternative management option. METHODS: We did a retrospective review of perioperative endoscopic retrograde cholangiopancreatographies (ERCPs) and sphincterotomies that were performed on patients in whom choledocholithiasis had been revealed on operative cholangiogram. RESULTS: Thirteen patients underwent perioperative ERCP. Nine had successful duct clearance, three had failure to cannulate, and one was converted to an open procedure due to an anatomical problem. The combined median operating time for both procedures was 75 min and hospital stay was 2.5 days. No pancreatitis was reported. CONCLUSION: Selective operative cholangiography and perioperative ERCP during laparoscopic cholecystectomy is a viable option for the simultaneous management of CBD and gallbladder stones. Moreover, it helps to avoid unnecessary normal ERCP. PMID- 11928031 TI - Laparoscopic cholecystectomy performed under regional anesthesia in patients with chronic obstructive pulmonary disease. AB - BACKGROUND: Laparoscopic cholecystectomy has been successfully performed using epidural anesthesia. We evaluated our experience with this surgical approach in high-risk patients. METHODS: We present the results of 29 patients with gallstones who, between 1998 and 1999, underwent laparoscopic cholecystectomy with epidural anesthesia. All but 1 patient had chronic obstructive pulmonary disease. RESULTS: All 29 surgeries were successfully completed via laparoscopy and with the patients under epidural anesthesia. No patient required endotracheal intubation during surgery or pain medication afterward. Postoperatively, 1 patient developed a wound infection and 3 patients developed urinary retention. At last follow-up (12 months postop), all patients were in good health. CONCLUSION: In this series, laparoscopic cholecystectomy was feasible under epidural anesthesia and it eliminated the need for postoperative analgesia. We believe that this approach should be considered for patients who require biliary surgery but who are not good candidates for general anesthesia due to cardiorespiratory problems. PMID- 11928030 TI - Does splanchnic ischemia occur during laparoscopic cholecystectomy? AB - BACKGROUND: Although it has been been shown that the elevation of abdominal pressure causes splanchnic ischemia, the pathophysiological changes that occur during laparoscopic cholecystectomy are still under investigation. Our aim was to evaluate the changes in nitric oxide (NO), malondialdehyde (MDA), and gastric intramucosal pH (pHi) during laparoscopic surgery to verify whether splanchnic ischemia had occurred. METHODS: Forty patients (11 men and 29 women) scheduled for cholecystectomy were randomly divided to undergo either open (OC) or laparoscopic cholecystectomy (LC). Repeated blood samples were collected for the measurement of NO and MDA concentrations. End-tidal carbon dioxide (ETCO2) levels were measured using a capnograph, and pHi was assessed using a tonometric nasogastric catheter. RESULTS: Although no significant change in pH was observed in the LC group during the procedure, it decreased significantly in the OC group and then returned to normal immediately after the procedure. ETCO2 did not change in the LC group but it increased in the OC group. Although it was not statistically significant, NO concentrations were slightly increased in the LC group. The same magnitude of increase was seen in MDA levels but it was not significant. Neither NO nor MDA levels changed in the OC group. CONCLUSION: LC has no adverse effects on gut perfusion. Even if the abdominal organs are hypoperfused during the procedure, compensates for this hypoperfusion so that it pulsatile NO secretion does not cause any harm. Since LC is a short procedure, the significance of these events for extended procedures still needs to be clarified. PMID- 11928032 TI - Sympathetic nervous system activity during laparoscopic and needlescopic cholecystectomy. AB - BACKGROUND: It is difficult to quantify the impact of surgical trauma on patients with conventional measurement techniques. We report our preliminary experience with the determination of sympathetic nervous system activity during laparoscopic and needlescopic cholecystectomy. METHODS: The electroconductivity of representative dermatomes (24 measuring points) was recorded prior, during, and after performing laparoscopic and needlescopic cholecystectomy on 40 consecutive patients according to the method of Nakatani (Ryodoraku). Fifty-five healthy adults served as a control group. RESULTS: Median age in the two groups was 52.4 and 54.1 years, respectively. Prior to the operation and on the 1st postoperative day, sympathetic nervous activity was equal in both groups. In the needlescopic patients, sympathetic activity was lesser at 30 min after the start of the laparoscopic procedure (p <0.05). CONCLUSIONS: The measurement of electroconductivity is a useful noninvasive technique for the evaluation of sympathetic nervous activity. As far as activation of sympathetic nervous system is concerned, the needlescopic technique was found to be superior. PMID- 11928033 TI - Functional impairment of the abdominal wall following laparoscopic and open cholecystectomy. AB - BACKGROUND: Subcostal incisions for open cholecystectomy (OC) denervate the right upper and middle abdomen. METHODS: Sensory and abdominal muscle function (Janda's muscle function test, Cybex 6000) was evaluated. Healthy volunteers (10 women, 12 men) were compared with 13 women and 12 men after OC and 11 women and 11 men after laparoscopic cholecystectomy (LC). For computed tomography (CT) studies, a spiral scanner was used. RESULTS: Of the OC patients, 21 (95.5%) complained of dys- and anesthesias below the incision. Of the LC patients, only nine (2.4%) reported anesthesias around the port sites. Normal subjects scored significantly higher in muscle strength (p <.01). LC patients scored higher than OC patients at 10 degrees and 20 degrees trunk flexion (p <.05). Depending on the flexion angles, the men in all groups developed 30-114% more muscle power than the women. Denervation of the abdominal muscles was confirmed by CT. CONCLUSION: OC reduced skin sensitivity and abdominal muscle function. Although the procedure is minimally invasive, LC also causes sensory and functional losses, albeit minor ones. PMID- 11928034 TI - Laparoscopic endobiliary stenting significantly improves success of postoperative endoscopic retrograde cholangiopancreatography in low-volume centers. AB - BACKGROUND: Occult common bile duct stones (CBDS) discovered during laparoscopic cholecystectomy with intraoperative cholangiography are most often managed by postoperative endoscopic retrograde cholangiopancreatography (ERCP). Expert endoscopists at high-volume centers achieve common bile duct cannulation in nearly all patients undergoing ERCP, but cannulation rates of less than 80% have been observed in low-volume centers. As many as 20% of patients with CBDS referred for postoperative ERCP in low-volume centers may require repeated attempts at ERCP, referral to a high-volume center, percutaneous transhepatic techniques, or reoperation for clearance of CBDS when postoperative ERCP fails. METHODS: Laparoscopic cholecystectomy with intraoperative cholangiography performed in 511 consecutive patients over 80 months at a community hospital showed occult CBDS in 66 patients (12.9%). Laparoscopic endobiliary stent placement was successful in 65 patients (98.5%). As part of an earlier study, 16 patients underwent laparoscopic common bile duct exploration with clearance of CBDS before stent placement. Laparoscopic endobiliary stent placement failed in one patient for whom CBDS were cleared with intraoperative ERCP. RESULTS: Initial postoperative ERCP was successful in clearing CBDS in all 65 patients (100%) with laparoscopically placed stents. During the same period, 611 patients underwent ERCP for various indications including CBDS (43%). Selective cannulation was achieved in 78% of all patients during initial ERCP. CONCLUSIONS: Laparoscopic endobiliary stent placement is an effective adjunct to the management of occult CBDS. Laparoscopic endobiliary stenting ensures selective cannulation during postoperative ERCP and eliminates the need for repeated attempts at ERCP, referral to specialty centers, use of transhepatic techniques, or reoperation for retained CBDS. Laparoscopic endobiliary stent placement for treatment of occult CBDS significantly improves the success of postoperative ERCP in low-volume centers and eliminates the morbidity and expense of repeated procedures. PMID- 11928035 TI - Peroperative analysis of the surgical procedure. AB - The increased technological complexity of surgery and the growing importance of quality assessment demand objective analysis of the surgical process. However, until now no standard method existed for analyzing the peroperative process. In this article, a methodology is discussed to describe and to analyze the surgical process. A method is given to measure the correctness and efficiency of task performance, protocols, and instruments used. In addition, reference values are defined so as to compare new instruments, alternative protocols, and the performance of new tasks with a standard. Finally, recommendations are given for improving new surgical tasks, the development of clinically driven instrument design, and new protocols. PMID- 11928036 TI - Laparoscopic lumbar sympathectomy for lower-limb disease. AB - BACKGROUND: The standard procedure for sympathectomy is open surgery. The oblique retroperitoneal approach is popular because it provides good visibility, albeit at the expense of requiring a long skin incision. Chemical sympathectomy has been introduced as a less invasive means of achieving sympatholysis; however, this method is also associated with a significant incidence of incomplete block and transient denervation. Laparoscopic surgery is a new approach that simplifies various surgical procedures. The aim of our report was to evaluate the benefits of endoscopic retroperitoneal surgery for lumbar sympathectomies. METHODS: Between March 1997 and April 2000, seven patients underwent laparoscopic lumbar sympathectomy in our department (all men, with an average age or 45.1 years). The predominant presenting symptoms were unilateral pain at rest and lower-extremity coldness. Symphaectomy was performed using a retroperitoneal approach on six patients and an anterior transperitoneal approach on one patient. After laparoscopic lumbar sympathectomy, skin thermometry was carried out on all patients. RESULTS: The postoperative skin temperature of the affected leg rose to 36.6 +/- 0.5 degrees C, as compared to 33.8 +/- 0.8 degrees C preoperatively. After laparoscopic lumbar sympathectomy, none of the patients complained of neuralgia. All patients achieved sustained symptomatic relief, and no major postoperative complications were noted. CONCLUSIONS: Lumbar sympathectomy can be performed laparoscopically. Currently, our standard technique is the retroperitoneal approach. More clinical experience and long-term follow-up will ultimately determine if this will become the procedure of choice. However, we believe that a learning period is necessary for this technique to be fully mastered. PMID- 11928037 TI - The suprapubic approach for laparoscopic appendectomy. AB - BACKGROUND: Because it produces superior cosmetic results, patients prefer laparoscopic appendectomy over open appendectomy. We developed two alternative laparoscopic routes of access to the abdominal cavity for appendectomy that use suprapubic incisions placed below the line of pubic hair. We then compared the results for these three different modes of access. METHODS: Operative characteristics, morbidity, outcome, and patient preference regarding three different approaches to laparoscopic appendectomy were compared in a retrospective study. In addition, a group of 24 healthy women were surveyed by questionnaire about their preferred technique and expected cosmetic results. RESULTS: Between January 1997 and August 2000, 149 patients underwent laparoscopic appendectomy and were assigned to undergo one of the three techniques. Operative results, morbidity, and hospital stay were similar. Twenty five percent of patients submitted to technique 1 (no suprapubic trocars) were satisfied with their method, vs 54% of patients with technique 2 (one suprapubic port, angled working trocars) and 100% of patients with technique 3 (two suprapubic parallel trocars). Almost all patients (92% of those who had technique 1 and 100% of those who had techniques 2 and 3) chose the standard laparoscopic access as the cosmetically least attractive method. All of the healthy controls we interviewed preferred technique 3. CONCLUSION: The placement of suprapubic trocars improves the surgeon's working position during laparoscopic appendectomy. A laparoscopic approach using two suprapubic trocars yields the best cosmetic results in the opinion of the majority of patients and healthy interviewees. PMID- 11928038 TI - Preoperative localization of indeterminate pulmonary nodules before videothoracoscopic resection. AB - BACKGROUND: The management of a solitary pulmonary nodule is the subject of debate and minimally invasive diagnostic approaches have low sensitivity for small peripheral nodules. We discuss the role of video-assisted thoracoscopic surgery (VATS) in the management of solitary pulmonary nodules (SPNs) < or = 1 cm performed with a preoperative computed tomography-guided wire localization. METHODS: Thirty-five selected patients underwent VATS resection for SPN, with localization by guide wire before surgery. RESULTS: Seven patients, after VATS exploration, underwent thoracotomy because of pleuropulmonary adhesions, depth or dimensions. Histological diagnosis was obtained in all procedures; there was no postoperative morbidity or morbidity. CONCLUSION: Preoperative computed tomography hook-wire localization is a suitable strategy for peripheral nodules < or = 1 cm in diameter. PMID- 11928039 TI - Diagnosing the occult contralateral inguinal hernia. AB - BACKGROUND: The incidence of bilateral inguinal hernias reported for total extra peritoneal (TEP) laparoscopic hernia repair, which reaches 45%, appears to be higher than that seen in studies of transabdominal laparoscopic and open repair. Given the unique ability of diagnostic laparoscopy to diagnose occult contralateral hernias (OCH) accurately, this study looked at how concurrent transabdominal diagnostic laparoscopy (TADL) would influence planned TEP repairs. METHODS: A prospective study oF 100 consecutive TEP cases was conducted. All patients had diagnostic laparoscopy via a 5-mm 45 degrees scope through an umbilical incision with 15 mmHg of pneumoperitoneum, followed by laparoscopic TEPrepair. A contralateral occult hernia was diagnosed and repaired if a true peritoneal eventration through the inguinal region was observed. RESULTS: Among the 100 patients, preoperative diagnosis suggested 31 bilateral hernias (31%), whereas TADL confirmed 25 bilateral hernias (25%). Of these 25 bilateral hernias, TADL confirmed 16 that had been diagnosed preoperatively (64%), but excluded 15 contralateral hernias that were incorrectly diagnosed (37%). Transabdominal diagnostic laparoscopy found nine OCHs, representing 36% of all bilateral hernias and 13% of the 69 preoperatively determined unilateral hernias. The preoperative physician examination false-negative rate for contralateral hernias was 36%, and the false-positive rate was 37%. In 26 cases (26%), TADL changed the operative approach. CONCLUSIONS: In this study, patients believed to have unilateral inguinal hernias had OCHs in 13% of cases when examined by TADL. The actual bilateral hernia incidence was 25%, with a 37% false-positive rate for preoperatively diagnosed bilateral hernias. The high rate of bilateral hernias reported by the TEP approach alone suggests that some OCH findings may be an artifact of the TEP dissection. However, failure to search for an OCH could result in up to 13% of patients subsequently requiring a second repair. Because some surgeons are concerned about unnecessary TEP dissection of the asymptomatic contralateral side, the approach described here may offer a solution to accurate diagnosis of the contralateral inguinal region during planned laparoscopic TEP hernia repair. PMID- 11928040 TI - Laparoscopic pericystectomy for liver hydatid cysts. AB - BACKGROUND: The laparoscopic approach for managing of liver echinococcosis is a controversial issue because of scarce experience worldwide. The aim of this report is to describe the technical details of our laparoscopic method and present our results. METHODS: Consecutive cases of liver echinococcosis managed by laparoscopic surgery are reported. Thoracic x-ray and abdominal ultrasound had been performed previously. The following aspects were considered as selection criteria: unique cyst located in segments III, IV, V, VI, and VIII; diameter less than 7 cm; and no evidence of infection or calcification. An evacuating puncture was performed, germinative membrane removed, and pericystectomy performed, which extirpated the pericystic structure with the surrounding liver parenchyma. Specimens were removed in a plastic bag through one of the ports. Surgical morbidity, hospital stay, time until return to work, and evidences of hydatid recurrence were measured. RESULTS: Surgery was performed on eight patients (5 women and 3 men) with a mean age of 44.9 years (range, 22-83 years) who had a liver hydatid cyst with a mean diameter of 6.6 cm (range, 5-7 cm). During a mean follow-up period of 30 months (range, 23-44 months), no morbidity or hydatid recurrence were verified. Hospital stay was 2 days in all cases, and return to work was within 15 days. CONCLUSION: This laparoscopic technique, applied with selective criteria, can be a useful alternative for treating patients with liver hydatidosis because its results are comparable with those for open surgery studies involving similar follow-up time. PMID- 11928041 TI - The influence of CO2 versus helium insufflation or the abdominal wall lifting technique on the systemic immune response. AB - BACKGROUND: Both laparoscopic and conventional surgery result in activation of the systemic immune response; however, the influence of the laparoscopic approach, using CO2 insufflation, is significantly less. Little is known about the influence of alternative methods for performing laparoscopy, such as helium insufflation and the abdominal wall lifting technique (AWLT), and the systemic immune response. METHODS: Thirty-three patients scheduled for elective cholecystectomy were randomly assigned to undergo laparoscopy using either CO2 or helium for abdominal insufflation or laparoscopy using only the AWLT. The postoperative inflammatory response was assessed by measuring the white blood cell count, C-reactive protein (CRP) and interleukin-6 (IL-6). The postoperative immune response was assessed by measuring monocyte HLA-DR expression. RESULTS: CRP levels were significantly higher 1 day after helium insufflation when compared with CO2 insufflation; however, no differences were observed 2 days after surgery. The AWLT resulted in significantly higher levels of CRP both 1 and 2 days after surgery when compared with either CO2 or helium insufflation. A small increase in postoperative IL-6 levels was observed in all groups, but no significant differences were seen between the groups. After both helium insufflation and AWLT a significant decrease in HLA-DR expression was observed, in contrast to the CO2 group. CONCLUSION: Carbon dioxide used for abdominal insufflation seems to limit the postoperative inflammatory response and to preserve parameters reflecting the immune status. These findings may be of importance in determining the preferred method of laparoscopy in oncologic surgery. PMID- 11928042 TI - Impact of different gases and pneumoperitoneum pressures on tumor growth during laparoscopy in a rat model. AB - BACKGROUND: The influence on intraperitoneal tumor growth of the choice of gas and pneumoperitoneum pressure during laparoscopy is still unknown. This study compared tumor growth after laparoscopy with different gases and pneumoperitoneum pressures in an immunodeficient model. METHODS: In an initial experiment, 60 nude rats were randomly allocated to undergo laparoscopy at different pneumoperitoneum pressures (gasless, 4 mmHg, or 8 mm Hg.) In a second experiment, 23 nude rats were randomly allocated to undergo laparoscopy with different gases (carbon dioxide or helium). Surgery was carried out 7 days after intraperitoneal injection of IGR-OV1 cells. The rats were killed 7 days after surgery. Tumor growth was assessed by the weight of the omental metastasis. For statistical analysis, we used analysis of variance (ANOVA). RESULTS: Mean omental weight was similar for all groups, regardless of the pneumoperitoneum pressure (p = 0.86) or the type of gas (p = 0.80). CONCLUSION: Physical parameters of gas have a limited impact on tumor growth. PMID- 11928043 TI - Pneumoperitoneum augmented tumor invasiveness is abolished by matrix metalloproteinase blockade. AB - BACKGROUND: Certain surgical strategies, including Helium (He) and carbon dioxide (CO2) insufflation in laparoscopy, have been shown to induce a hypoxic environment. This may have a significant effect on the invasive capacity of tumor cells and may be a factor in the incidence of port-site metastases seen in patients following laparoscopic resection for malignancy. METHODS: A colon adenocarcinoma cell line (SW1222) was exposed to an in vitro pneumoperitoneum of CO2 or He at 3 mmHg or left in normal growth conditions (control). After a 4-hour exposure to an in vitro pneumoperitoneum, the ability of the cells to invade through 8.0-microm Transwell filters coated with Matrigel was analyzed by colorimetric MTS assay and by direct staining of the filters. The effect of the addition of a known blocker of matrix metalloproteinases (MMPs), 1,10 phenanthroline (1,10-P), was investigated. RESULTS: Cells exposed to an in vitro pneumoperitoneum demonstrate significantly increased invasive capacity compared to the control set, without loss of viability (He vs control, p <0.001; CO2 vs control, p <0.001). This augmented capacity is abolished by the addition of 1,10 P (p <0.01). CONCLUSION: Exposure of a colonic adenocarcinoma cell line to either a CO2 or He pneumoperitoneum causes an increase in tumor cell invasiveness, which is abolished by the presence of a known inhibitor of MMPs. This suggests that MMPs have an important role in the metastatic potential of tumors exposed to a hypoxic operative environment. PMID- 11928044 TI - Combined endoscopic intragastral resection of a posterior stromal gastric tumor using an original technique. AB - Gastric stromal tumors are solitary, usually asymptomatic, lesions that can bleed, become obstructive, or even degenerate into malignant neoplasms. Therefore, their surgical excision is recommended. We report a technique for the successful resection of a stromal tumor of the posterior gastric wall using a transgastric approach. After the creation of a 12 mmHg pneumoperitoneum using a three-trocar technique, a 2-cm gastrostomy was performed; an 18-mm trocar was then positioned in the gastric lumen and secured with a pursestring suture. Next, an intragastric wedge resection of the posterior gastric wall was carried out under endoscopic guidance. Finally, the anterior gastric wall was closed using a linear stapler. Histopathological analysis showed a benign spindle cell tumor, which was excised in toto. Patient recovery was uneventful. This report supports previous data showing the feasibility of a laparoscopic transgastric approach for the resection of stromal tumors of the posterior gastric wall. It also underscores the synergy of laparoscopic and endoscopic procedures in minimally invasive gastric surgery. PMID- 11928045 TI - Laparoscopic removal of an ingested pin migrating into the liver. AB - Most ingested foreign bodies pass through the gastrointestinal tract without giving rise to complications. The possibility of penetration of the intestinal tract, by sharp, pointed objects, however, necessitates careful and continued observation. If such objects become lodged in a narrow segment of the gastrointestinal tract, perforation may occur. The resulting morbidity depends on the further route of the penetrating object and whether septic sequelae ensue. Although foreign bodies may migrate to almost any intraabdominal organ, perforation of the duodenum and migration into the liver are extremely rare. A case of a woman who unknowingly ingested a pin that perforated the duodenum causing only few acute symptoms is presented. Biliary tract pathology was suspected, but ultrasound examination ruled it out. Computed tomography of the abdomen showed a pin thrust into the liver, with the head of the pin in the wall of the duodenum. Traditional surgical treatment requires laparotomy for foreign body removal. In the reported case, the pin was removed laparoscopically. The postoperative course was uneventful, and the patient left the hospital on day 2 after the procedure. With laparoscopic approach for removal of penetrating intestinal foreign bodies, laparotomy and its attending complications are avoided. This approach is less invasive, has a beneficial impact on postoperative pain, produces a better cosmetic result, and offers a faster return to normal activities. Hospital stay and costs also are reduced. PMID- 11928046 TI - Acute gastric volvulus related to adhesions after laparoscopic fundoplication. AB - Laparoscopic antireflux procedures currently are considered to be as safe, cost efficient, and effective as the techniques used previously in antireflux surgery, although life-threatening complications after antireflux surgery have been reported with both open and laparoscopic fundoplication. We report the case of a 38-year-old man who presented with acute abdominal pain and vomiting 8 months after a laparoscopic Nissen-Rosetti fundoplication. The diagnosis of gastric volvulus was suspected. The endoscopic examination showed a proper location of the fundoplication and features consistent with early gastric ischemia. Rotational maneuvers failed to untwist the volvulus. Emergency surgery was performed, involving a conventional laparotomy. At surgery, the volvulus was found related to a thick adhesion between the opening of the xiphoidal cannula and the hilus hepatis. The section of the adhesion allowed immediate untwisting of the volvulus. Despite a general assumption that laparoscopic procedures will reduce the incidence of postoperative adhesion formation, it should be kept in mind that laparoscopic techniques do not preclude the deposit of unusual foreign microbodies, which is an admitted mechanism of adhesion formation. PMID- 11928048 TI - Small bowel volvulus resulting from a congenital band treated laparoscopically. AB - In recent years, laparoscopy has dramatically changed the approach to the patient with acute abdominal pain. We report the case of a patient with small bowel volvulus caused by a congenital band binding the greater omentum to the mesentery, which was promptly diagnosed and treated using laparoscopy. Early intervention averted irreversible ischemic lesions of the intestine and the need for bowel resection. With the routine use of laparoscopy in the setting of acute abdominal pain, rare affections can be easily diagnosed and effectively treated. PMID- 11928047 TI - Gastrocolocutaneous Fistula as a complication of peg tube placement. AB - A gastrocolocutaneous fistula after placement of a percutaneous endoscopic gastrostomy (PEG) tube is a rare complication. This fistula is defined as an epithelial connection between the mucosa of the stomach, colon, and skin. We present an interesting case in which a PEG tube was placed inadvertently through the colon. During a subsequent fever workup, an abdominal computed tomography scan suggested that the feeding tube had traversed the colon. Gastrograffin enema and colonoscopy confirmed the location of the misplaced feeding tube. We review the clinical presentation, diagnosis, errors in procedural technique, and treatment of gastrocolocutaneous fistulas. PMID- 11928049 TI - Perforation of the ileum due to foreign body treated laparoscopically. AB - Laparoscopy has gained widespread acceptance in the setting of acute abdominal pain. We report the case of a patient with acute abdominal pain in the right lower quadrant that proved to be due to a fish bone perforation of the ileum at laparoscopy. The fish bone was retrieved and the perforation closed laparoscopically. Laparoscopy is a powerful diagnostic and therapeutic tool and should be used routinely in patients with acute abdominal affections. PMID- 11928050 TI - Laparoscopic resection of a juxtaglomerular cell tumor of the right kidney. AB - Juxtaglomerular cell tumor is an extremely rare neoplasm of the kidney that causes blood hypertension. A 45-year-old man with persistent hypertension was referred to our department because of a solid mass of unclear origin (kidney? colon?) located in the right mesorenal region that had been detected by Computed tomography (CT) scan, ultrasonography, and colonoscopy. Serum levels of renin, plasma renin activity (PRA), and aldosterone were all normal. Operatively, four 10/12-mm ports were placed, ultrasonography confirmed a well-encapsulated lesion of the kidney. Ultrasonic shears and cautery were used to resect the lesion, including 5 mm of free renal tissue. Postoperatively, a prompt normalization of blood pressure was observed. The pathological findings showed a juxtaglomerular cell tumor. The definitive diagnosis of juxtaglomerular cell tumor is made on the basis of laboratory findings, and imaging studies, Its definitive treatment is surgical excision which can be accomplished successfully with minimally invasive surgery. PMID- 11928052 TI - Laparoscopic herniorrhaphy. PMID- 11928051 TI - The technique of laparoscopically assisted total gastrectomy with jejunal interposition for early gastric cancer. AB - BACKGROUND: In recent years, laparoscopic gastrectomy has been applied to the treatment of gastric cancer in Japan. However, there are few reports of laparoscopic or laparoscopically assisted total gastrectomy in the treatment of gastric cancer because of the difficulty of the surgical technique. Laparoscopically assisted total gastrectomies with jejunal interpositions were performed on four patients with early gastric cancer located in the upper portion of the stomach. METHODS: Four surgical ports were inserted into the abdomen. The stomach was lifted to the abdominal wall using newly developed retraction tubes. Gastric arteries were divided using ultrasonically activated coagulating shears and ligated with ligation forceps. Following these steps, a total gastrectomy reconstruction was performed by jejunal interposition through a small transverse laparotomy. An esophagojejunostomy and a jejunoduodenostomy were made with circular staplers. RESULTS: The mean operating time and blood loss were 246 min and 236 ml, respectively. The operations were performed without serious complications. All patients were pain free and ambulatory after the laparoscopically assisted total gastrectomy, and the mean postoperative hospital stay was 16 days. CONCLUSION: We successfully performed laparoscopically assisted total gastrectomies in a relatively short period of time. When patients are carefully selected, the laparoscopic procedure can be curative and minimally invasive as a treatment for early gastric cancer. PMID- 11928053 TI - Impact of randomized trials regarding endoscopic inguinal hernia repair in The Netherlands. PMID- 11928054 TI - Iatrogenic thoracic migration of the stomach complicating laparoscopic Nissen, fundoplication. PMID- 11928057 TI - [The "new period" in medical college]. PMID- 11928058 TI - [Secondary prevention of coronary heart disease. Results from EuroASPIRE I and II in the region of Munster, Germany]. AB - BACKGROUND: EuroASPIRE (European Action on Secondary Prevention through Intervention to Reduce Events) is a European multicenter study on secondary prevention in patients with coronary heart disease (CHD). The first cross sectional survey was undertaken in 1995/96 among 3569 patients from nine countries. A second cross-sectional survey (EuroASPIRE II) was conducted in 1999/2000 among 5556 patients from 15 countries to evaluate among others whether coronary prevention had improved since the first. METHODS: The present study was conducted in the region of Munster, Germany, as part of the EuroASPIRE study. Consecutive patients, men and women up to 70 years of age with established CHD, were identified retrospectively. A total of 392 (EuroASPIRE I) and 402 (EuroASPIRE II) patients participated. Information on cardiovascular risk factors, lifestyle and medication were obtained through medical records, interviews and examinations. RESULTS: Both studies demonstrate a high prevalence of risk factors among CHD patients. At the time of the interview more than 60% of the patients in both surveys had two or more risk factors. The comparison of EuroASPIRE I and II reveals a substantial decrease of 20% in the prevalence of hypercholesterolemia, but an increase in the prevalence of hypertension and obesity. ACE-inhibitors, betablockers and lipid lowering drugs, especially statins, were used more frequently in EuroASPIRE II. CONCLUSIONS: We conclude that secondary prevention of CHD in the region of Munster like in the other European study regions is less than optimal and has not substantially improved between 1996 and 2000. Potential reasons are discussed. PMID- 11928059 TI - [Cardiac resynchronization in heart failure]. PMID- 11928060 TI - [Treatment of heart failure with beta-receptor-blockers]. PMID- 11928062 TI - [PTCA or bypass-surgery in patients with renal failure and diabetes - pro surgery]. PMID- 11928061 TI - [Percutaneous coronary intervention in patients with diabetes]. PMID- 11928063 TI - [Minimally invasive coronary surgery - pro]. PMID- 11928064 TI - [Minimally invasive coronary surgery - contra]. PMID- 11928065 TI - [How is prevention of thrombosis in paroxysmal absolute tachyarrhythmia managed?]. PMID- 11928066 TI - Delay of growth and development in children with bronchial asthma, atopic dermatitis and allergic rhinitis. AB - The elevated incidence of short stature (body height < (-)x - 2s), skeletal retardation and delayed puberty in children with bronchial asthma or atopic dermatitis is generally attributed to the severity of the disorder. However, a series of findings indicate a causal influence of the atopy and the existence of atopic skeletal retardation per se.The observation that children with atopic disorders, whether bronchial asthma, atopic dermatitis or allergic rhinitis, exhibit a rate of short stature that is twice to five times higher than normal indicates atopic and thus genetically determined influences. The elevated prevalence of short stature associated with allergic rhinitis is especially significant, as this disorder cannot be included among the severe chronic disorders. The fact that skeletal retardation is more prevalent in boys than in girls by a ratio of about 2:1 and that a significantly more marked retardation of bone maturation is found in atopic in comparisons with non-atopic asthmatics also lend support to this postulation. The clinical relevance of atopic growth retardation is also supported by the close interaction of pathophysiological basal mechanisms of bone metabolism and the atopy status. Thus the local growth factor prostaglandin E(2) (PGE(2)), which is important for bone metabolism, is also a messenger substance for the immediate and late allergic reaction. The platelet-activating factor (PAF), as one of the strongest mediators in the pathogenesis of allergic disorders, influences the PGE(2) synthesis in the osteoblasts. These relationships show that atopy-dependent imbalances in the complex system of local and systemic growth factors can certainly lead to disturbance of skeletal maturation which may delay growth and development in atopic children. In order to verify these assumptions it is necessary to research the interaction of local growth factors (particularly the roles of PGE(2), PAF and IGF I) in the skeletons of children of short stature suffering from atopic disorders. This should also include the possible effects on the overall hormonal factors influencing bone maturation. Atopy should be included in the differential diagnosis programme to clarify growth and development disturbances. PMID- 11928067 TI - Pro12Ala polymorphism in the peroxisome proliferator-activated receptor-gamma2 (PPARgamma2) is associated with higher levels of total cholesterol and LDL cholesterol in male caucasian type 2 diabetes patients. AB - The peroxisome proliferator-activated receptor-gamma2 (PPAR(gamma2)) represents the transcriptional master regulator of adipocyte differentiation and therefore has been suggested as candidate gene for the pathogenesis of obesity, type 2 diabetes and related metabolic disorders. Aim of our study was to determine the frequency of a missense point mutation within exon 2 of PPAR(gamma2), Pro12Ala, and its possible association with metabolic parameters as well as diabetic retinopathy (in a population-based sample of 560 (318 male ad 242 female) type 2 diabetic patients. Subsequent to genomic PCR amplification, the Hpa-II RFLP analysis was used for genotyping. RESULTS: 436 (77.9%) subjects were homozygous for the wildtype allele (Pro/Pro), 118 (21.1%) were heterozygous (Pro/Ala) and 6 (1.1%) were homozygous for the mutated allele (Ala/Ala). Genotype frequency was calculated to be 0.81 for the wildtype and 0.19 for the mutated allele. These frequencies did not differ from non-diabetic cohorts examined earlier. In contrast to females, total cholesterol and LDL-cholesterol were significantly higher in males (Total cholesterol: 281.8 +/- 51.3 vs 253.1 +/- 49.8 mg/dl, p < 0.0001; LDL-cholesterol: 182.0 +/- 49.2 vs 155.6 +/- 42.0 mg/d, p < 0.0001) in the presence of the mutated allele as compared to the wildtype subgroup. No differences were found with respect to BMI, HbA1c, blood pressure and serum levels of leptin nor to prevalence of retinopathy. Pro12Ala polymorphism of PPAR(gamma2) gene is not associated with diabetic retinopathy but is associated with dyslipidemia in male type 2 diabetic patients. PMID- 11928069 TI - Homocysteine and steroids levels in metformin treated women with polycystic ovary syndrome. AB - Risk for atherosclerosis is increased in women with polycystic ovary syndrome (PCOS). Homocysteine (Hcy) is one of the independent risk factors for ischemic heart disease. We examined the effect of metformin (M) treatment on Hcy levels, steroids and glycide tolerance in PCOS. MATERIAL AND METHODS: 9 women with PCOS (defined as hyperandrogenemia and chronic anovulation); age 20 +/- 3.8 yrs, BMI 28.1 +/- 6.5 kg/m(2); examined in the follicular phase of spontaneous menstrual cycle before and after 27 +/- 4 weeks of treatment with M 1000 mg/day. The plasma concentrations of Hcy, DHEA, DHEA-S, cortisol (F), allopregnanolone (HPO), 17OHpregnenolone (17OHPl), insulin (I) and blood glucose (G) before and after the course of M were measured. RESULTS: After the course of M, Hcy significantly increased (10.1 +/- 2.6 to 13.4 +/- 5.1 micromol/l, p < 0.05.). There was no significant change in levels of I, HPO, F, DHEA-S and 17OHPl. DHEA levels increased significantly (from 26.9 +/- 15.7 to 44.4 +/- 24.6 nmol/l, p < 0.05). A borderline significant trend towards reduction in waist-hip ratio was seen (from 0.986 +/- 0.042 to 0.951 +/- 0.085; p < 0.06). CONCLUSIONS: Treatment with metformin in women with PCOS can lead to the increase in homocysteine levels--a risk factor for atherosclerosis. PMID- 11928070 TI - Thermal destabilization of ovarian LH/hCG receptors by negatively charged lipids. AB - The stabilizing effect of BSA on the rat ovarian LH/hCG receptor was analyzed by thermal perturbation technique. Thermal destabilization of the receptor with arachidonic acid along with digestion of membrane with phospholipase A2 and reversal of these effects when BSA was used as fatty acids scavenger, may indicate that free fatty acids are responsible for instability of the LH/hCG receptor. This destabilizing effect may be caused by the presence of a net negative surface charge provided by fatty acids. This presumption was corroborated by the reconstitution of delipidated LH/hCG receptor into proteoliposomes. Delipidated receptor lost to a great extent its binding activity and thermal stability. The receptor was fully reactivated by the reconstitution into proteoliposomes with neutral phosphatidylcholine but not with negatively charged phosphatidylserine and phosphatidylglycerol. Thermal inactivation of the LH/hCG receptor by delipidation was entirely inverted by treatment with phosphatidylcholine but the presence of negatively charged phospholipids did not change the heat inactivation profile of hCG-binding sites. PMID- 11928068 TI - Transport of insulin-like growth factor-I across endothelial cell monolayers and its binding to the subendothelial matrix. AB - Cultured human umbilical vein endothelial cells (HUVEC) were used as a model to study transendothelial IGF-I transport, and its deposition into the extracellular matrix (ECM). Specific binding of (125)I-IGF-I to HUVEC monolayers was demonstrated, which was inhibited by aIR-3, a specific antibody directed against the IGF-I receptor. ECM-associated (125)I-IGF-I was approximately 10% of cell bound IGF-I at 22 degrees C, and increased 4.5-fold at 37 degrees C, indicating that endothelial metabolism is required for the transport. However, neither monensin and cytochalasin B, both of which block endocytosis, nor aIR-3 did inhibit transport of (125)I-IGF-I into the ECM. In order to characterize IGF-I binding to the subendothelial ECM, HUVEC were removed nonenzymatically by treatment with Triton X-100 and ammonia. Specific, saturable binding of (125)I IGF-I to the isolated ECM was observed, which was protease-sensitive. Antibodies directed against vitronectin inhibited IGF-I binding to the matrix by 35%, while antibodies directed against other ECM proteins had no significant influence on IGF-I binding. Using radioimmunoassays the IGF binding protein-2 was detected in the ECM, while IGFBP-1 and IGFBP-3 were below the detection limits. In order to evaluate functional aspects of IGF-I binding to the matrix, HUVEC were incubated under serum-free conditions in the absence and presence of IGF-I. Under serum free conditions 48% of cells rounded up and started to detach after 2 hours incubation, while only 23% of the cells started to detach in the presence of IGF I. These data indicate that IGF-I is transported via a paracellular route across endothelial cells, and becomes bound to the subendothelial ECM. Vitronection seems to be involved in binding of IGF-I to the ECM. ECM-associated IGF-I might play a role in endothelial cell survival and stability. PMID- 11928071 TI - Ratio of serum aldosterone to plasma renin concentration in essential hypertension and primary aldosteronism. AB - The ratio of serum aldosterone to plasma renin activity (PRA) has been proposed as sensitive screening method in the diagnosis of primary aldosteronism under random conditions. However, the method for determination of renin activity is hampered by the necessity of ice cooling during storage and transport. The present study was therefore conducted to examine the ratio of serum aldosterone to plasma renin concentration (ARR) and its usefulness in diagnosis of primary aldosteronism under ambulatory conditions and given antihypertensive medication. 146 patients with arterial hypertension who consecutively attended the outpatient clinic were studied prospectively. Patients with secondary hypertension besides primary aldosteronism were not included in the series. 37 normotensive patients served as control. Also, 17 patients with known primary aldosteronism were retrospectively examined. Among the hypertensive group 2 patients with Conn's syndrome were newly detected (1.4%). ARR was 7.92 +/- 6.04 [pg/ml]/[pg/ml] in normotensive controls (range from 2.03 to 26.98), 14.61 +/- 18.50 [pg/ml]/[pg/ml] in patients with essential hypertension (n = 144, range from 0.41 to 115.45) and 155.92 +/- 127.84 [pg/ml]/[pg/ml] in patients with primary aldosteronism (n = 19, range from 6.75 to 515). 17 of the 19 patients with Conn's syndrome had an ARR of more than 50. Under ongoing drug treatment this represents a sensitivity of 89% and a specificity of 96%. Sensitivity decreased to 84% and specificity increased to 100% when a second criteria (aldosterone > or = 200 pg/ml) was included. In summary, ARR using renin concentration is a useful screening parameter for primary aldosteronism. PMID- 11928072 TI - Abnormal pentagastrin response in a patient with pseudohypoparathyroidism. AB - The case of a 25 year old female patient with pseudohypoparathyroidism type I (PHP) and hypercalcitoninaemia is reported. She was referred to our clinic because of recurrent hypocalcaemia associated with paraesthesias and muscle cramps. She had no signs of Albright hereditary osteodystrophy (AHO), a normal mental status and no family history of hypocalcaemia or any other endocrine disease. Considering the laboratory results with hypocalcaemia, hyperphosphataemia, normal vitamin D and normal creatinine with an extraordinary elevated PTH we diagnosed pseudohypoparathyroidism type I. She had delayed pubertal development with menarche in the age of 20 and hypothyroidism with an atrophic thyroid since she was 22 years old. Calcitonin (CT) was increased and the performed pentagastrin test showed an excessive CT-response with a peak of 725 pg/ml after 2 min. Up to now there are only three reports of patients with PHP and hypercalcitoninaemia. An abnormal pentagastrin response is known to be a specific marker for medullary thyroid carcinoma, but there were no signs of any malignant disease, even after one year of follow-up. The most reasonable cause for the pathological pentagastrin response might be chronic hypocalcaemia. When interpreting a pathological pentagastrin test in a patient with PHP the specifity of the test might be diminished and a careful observational strategy might be appropriate. PMID- 11928073 TI - Determinants of early carotid atherosclerosis progression in young patients with type 1 diabetes mellitus. AB - The results of cross-sectional studies addressing early preintrusive atherosclerosis in type 1 diabetic patients are conflicting. In an observational longitudinal study we determined the course of carotid artery intima-media thickness (IMT) over a period of 2.5 years in mean. A total of 102 patients with type 1 diabetes mellitus (age < or = 40 years, diabetes duration > or = 2 years at baseline examination) who were participants of the baseline examination was studied again in a follow-up. HbA1c, albumin excretion rate (AER), lipids, systolic and diastolic blood pressure, retinopathy, and current smoking status were assessed at baseline and follow-up. The IMT of the common carotid artery was measured by high-resolution ultrasound, the maximum IMT was evaluated. The annual progression rate (APR) was calculated from the difference between baseline and follow-up IMT reading and the time between both examinations. The follow-up IMT was significantly higher, compared to the baseline measurement: 0.65 +/- 0.18 vs. 0.57 +/- 0.14 mm (p < 0.001), the mean APR was 0.033 mm/year. The APR was correlated with age (r = 0.337, p < 0.01), diabetes duration (r = 0.252, p < 0.05), hypertension (r = 0.225, p < 0.05), and systolic blood pressure (r = 0.281, p < 0.05) at the baseline examination. Comparing subgroups, defined according to APR tertiles, with no IMT progression (first tertile, mean APR - 0.012 mm/year), mild progression (second tertile, mean APR 0.037 mm/year), and advanced progression (third tertile, mean APR 0.088 mm/year), patients with advanced progression significantly (p < 0.05) more often had hypertension and nephropathy than subjects with mild progression. In a multiple linear regression analysis, the changes of plaque frequency and of the nephropathy status between baseline and follow-up examinations were independent predictors of the APR. PMID- 11928074 TI - Vascular and coagulation disorders of the liver. PMID- 11928075 TI - Hepatic vein thrombosis (Budd-Chiari syndrome). AB - Hepatic vein thrombosis is caused by one or several thrombogenic conditions, of which myeloproliferative disorders are the most frequent. Thrombosis and its fibrous sequelae can affect the veins diffusely or locally. Severity is determined by the extent and velocity of the thrombotic process. Development of venous collateral vessels is an important compensatory mechanism. Some patients can be totally free of symptoms. Major complications are intractable ascites, liver insufficiency, and gastrointestinal bleeding. Diagnosis can be made via ultrasonography or magnetic resonance imaging in a majority of cases. The main prognostic factors are age, Child-Pugh score, and response of ascites to diuretics. Medical therapy includes control of causal factors, anticoagulation, and nonspecific treatment of complications. Procedures aiming to restore outflow of hepatic blood are indicated in patients with uncontrolled manifestations. Percutaneous angioplasty, followed by portosystemic shunt (including a transjugular intrahepatic portosystemic stent shunt) and eventually liver transplantation can be proposed in a graded manner. The current 10-year survival rate is about 75%. PMID- 11928076 TI - Inferior vena cava thrombosis at its hepatic portion (obliterative hepatocavopathy). AB - The Budd-Chiari syndrome was primarily described as hepatic vein thrombosis within the liver, but it now includes inferior vena cava (IVC) thrombosis and other conditions that cause hepatic vein outflow obstruction. This author and several others maintain that primary hepatic vein thrombosis and primary IVC thrombosis represent two different clinical disorders. Primary thrombosis of the IVC most commonly occurs in its hepatic portion, which seems to be predisposed to thrombosis and has been called membranous obstruction of IVC, because the thrombus organizes into a fibrous and frequently membranous occlusion of the IVC. The hepatic vein orifices are affected to varying degrees, resulting in congestive liver damage. The cause of IVC thrombosis may be a hypercoagulable state such as coagulation factor deficiency and myeloproliferative disorders, but is more often idiopathic. In Nepal, it is endemic with a suspected association with infections. To consider IVC thrombosis and the congestive liver damage as a disease entity, this author proposes the term obliterative hepatocavopathy, separate from hepatic vein thrombosis. Clinically obliterative hepatocavopathy is less severe in its acute phase compared with hepatic vein thrombosis, but it aggravates occlusion of hepatic vein orifices with recurrent thrombosis. Primary hepatic vein thrombosis and obliterative hepatocavopathy display different hemodynamics of the hepatic veins, IVC, and portal vein; dilatation of the subcutaneous veins in the body trunk is more pronounced in obliterative hepatocavopathy because the ascending lumbar vein becomes the major collateral route. Congestive liver cirrhosis develops after a long clinical course that may be complicated by hepatocellular carcinoma. PMID- 11928078 TI - Extrahepatic portal vein obstruction. AB - Extrahepatic portal vein obstruction (EHPVO) is an important cause of noncirrhotic portal hypertension, especially in Third World countries. The etiology and clinical presentation are different in children and adults. The portal vein is transformed into a cavernoma, resulting in portal hypertension and oesophagogastic varices. In addition, extensive collateral circulation develops, involving paracholecystic, paracholedochal and pancreaticoduodenal veins resulting in formation of ectopic varices, and portal biliopathy. Besides variceal bleeding, which is the commonest presentation, patients may have symptomatic portal biliopathy, hypersplenism, and growth retardation. Although the liver may appear normal, functional compromise develops in the long term. Variceal bleeding in EHPVO can be successfully managed by endoscopic obliteration of varices, which has low morbidity but requires repeated visits, or by portosystemic shunt surgery, which provides good control of bleeding, possibly helps growth retardation, hypersplenism, and protects against future development of portal biliopathy but is associated with surgical mortality and is sometimes not feasible due to nonavailability of a satisfactory vessel. PMID- 11928077 TI - Toxic injury to hepatic sinusoids: sinusoidal obstruction syndrome (veno occlusive disease). AB - The term veno-occlusive disease of the liver refers to a form of toxic liver injury characterized clinically by the development of hepatomegaly, ascites, and jaundice, and histologically by diffuse damage in the centrilobular zone of the liver. The cardinal histologic features of this injury are marked sinusoidal fibrosis, necrosis of pericentral hepatocytes, and narrowing and eventual fibrosis of central veins. Recent studies suggest that the primary site of the toxic injury is sinusoidal endothelial cells, followed by a series of biologic processes that lead to circulatory compromise of centrilobular hepatocytes, fibrosis, and obstruction of liver blood flow. Thus we propose a more appropriate name for this form of liver injury--sinusoidal obstruction syndrome. This review encompasses historical perspectives, clinical manifestations of sinusoidal obstruction syndrome in the setting of hematopoietic cell transplantation, histologic features of centrilobular injury, and a discussion of the pathophysiology of sinusoidal injury, based on both animal and clinical investigations. PMID- 11928079 TI - Idiopathic portal hypertension and its pathology. AB - Idiopathic portal hypertension (IPH) is a disorder of unknown etiology, clinically characterized by portal hypertension (varices and portosystemic collateral vessels), splenomegaly, and anemia (hypersplenism). A similar disorder is called noncirrhotic portal fibrosis in India, and hepatoportal sclerosis seems to be the counterpart in the United States. This disease is uncommon in developed countries. Middle-aged women are more prone to IPH in Japan. The liver has no cirrhosis or pseudonodule formation, and the principal pathologic changes are considerable portal fibrosis, devastation of intrahepatic terminal portal radicles, and parenchymal atrophy of the liver secondary to portal malperfusion. The characteristic portal hemodynamics include intrahepatic presinusoidal portal hypertension, increased splenic and portal vein blood flow, and increased intrahepatic portal resistance. The prognosis is generally good depending on the management of bleeding varices. Although the etiology is obscure, certain immunologic abnormalities seem to play an etiologic role in Japanese patients, and the incidence has markedly declined in recent years in Japan, indirectly suggesting a role of infection. The theory that IPH represents an undiagnosed intrahepatic portal vein thrombosis is refuted. PMID- 11928080 TI - Pathomorphology of esophageal and gastric varices. AB - In this article, the gross pathology of varices and supplying veins are described comparing esophageal varices and varices of the cardia and fundus of the stomach. The angioarchitecture of the lower esophagus is such that normally very thin parallel veins in the lamina propria mucosae in the palisade zone become enlarged in portal hypertension and join the few larger submucosal veins to form esophageal varices. Enlarged parallel veins come to pile up and join the submucosal veins at an acute angle, rendering this area vulnerable to rupture. Most ruptures occur in this critical area. The basic differences between esophageal and gastric varices are the layers in which the varicose veins form: the lamina propria mucosae and submucosa in the esophageal varices and the submucosa in gastric varices. While cardiac veins and varices are continuous with esophageal varices, fundic varices develop independently as part of a splenogastrorenal shunt that runs through the stomach wall, having rare communications with other veins. The fundic varix is so large in caliber that when it ruptures, the muscularis mucosae and lamina propria are penetrated with massive bleeding. The treatment of varices calls for complete thrombosis of all varicose veins, and merits and demerits of available treatment modalities are discussed based on autopsies from the pathologic point of view. Because of the large size, the management of fundic varices is difficult, and the new technique called balloon-occluded retrograde transvenous obliteration for occluding fundic varices is discussed. PMID- 11928081 TI - Coagulation disorders in liver disease. AB - The liver plays a central role in the clotting process, and acute and chronic liver diseases are invariably associated with coagulation disorders due to multiple causes: decreased synthesis of clotting and inhibitor factors, decreased clearance of activated factors, quantitative and qualitative platelet defects, hyperfibrinolysis, and accelerated intravascular coagulation. The bleeding tendency accounts for increased risk of morbidity and mortality in patients with liver disease undergoing diagnostic or therapeutic invasive procedures. Peculiar coagulation disorders are prevalent in patients with acute fatty liver of pregnancy or undergoing liver transplantation. Emerging evidence shows that sepsis further impairs hemostasis in patients with liver cirrhosis bleeding from esophageal varices. Thrombotic events, even if rare in cirrhotic patients, occur mainly in the portal and mesenteric veins. The therapeutic approach to coagulative disorders is also discussed. PMID- 11928082 TI - A 25-year-old man with a large hepatic tumor and multiple nodular lesions. AB - A 25-year-old man without a prior history of liver disease presented with an 18 cm tumor of the right hepatic lobe, which was associated with multiple nodular lesions in the remaining parenchyma. The histologic and immunohistochemical features of the neoplasm were those of a poorly differentiated leiomyosarcoma with epithelioid features. The nodular lesions measured up to 4 cm in greatest dimension and had gross and microscopic features of focal nodular hyperplasia. This case suggests that multiple focal nodular hyperplasia may occasionally represent a parenchymal reaction to abnormal blood supply developing within the liver as a consequence of an enlarging malignant tumor. PMID- 11928083 TI - A 32-year-old man with hepatitis C who developed progressive liver failure after a bone marrow transplant. PMID- 11928084 TI - The "S" word. PMID- 11928085 TI - Tracheotomy versus endotracheal intubation for airway management in deep neck space infections. AB - PURPOSE: The aim of the present study was to compare costs and outcomes for patients with deep neck infections who were treated with either tracheotomy or endotracheal intubation. MATERIALS AND METHODS: The charts of 85 patients with deep neck space infections were retrospectively studied. Requirements for inclusion in the study were incision and drainage in the operating room, involvement of more than 1 deep anatomic space, impending airway compromise, and maintenance of a postoperative artificial airway. The 85 patients were divided into 2 groups based on the type of airway used for treatment. Group 1 (n = 34) included patients who received a tracheotomy, and group 2 (n = 51) included patients whose airways were maintained with endotracheal intubation until the swelling had resolved sufficiently for extubation. RESULTS: Patients in group 1 had a shorter overall hospital stay (4.8 vs 5.9 days, NS) and spent less time in the intensive care unit (1.1 vs 3.1 days, P <.05). The overall incidence of complications was 6% for group 1 and 10% for group 2. The rate of complications secondary to loss of airway was 3% for group 1 and 6% for group 2. Average costs associated with intensive care resources were 5 times greater and overall hospital stay costs were 60% greater for group 2. CONCLUSIONS: Although both methods of airway control are useful and have a unique set of complications, the use of tracheotomy allowed earlier movement to a noncritical care unit and was associated with fewer intensive care costs and less overall cost of hospitalization. Tracheotomy may therefore provide better utilization of critical care resources in this group of patients. PMID- 11928087 TI - Concomitant temporomandibular joint and orthognathic surgery: a preliminary report. AB - PURPOSE: In the present study, we evaluated the outcome of concomitant temporomandibular joint (TMJ) and orthognathic surgery in patients with TMJ articular disc dislocation and coexisting dentofacial deformities. PATIENTS AND METHODS: The records of 70 patients treated with TMJ articular disc-repositioning surgery and concomitant orthognathic surgery (double jaw or only mandibular surgery) were retrospectively evaluated. Patients were divided into the following 3 groups: group 1 patients had mandibular advancement, group 2 patients had mandibular setback, and group 3 patients had a mandible that remained in the original position. Lateral cephalometric radiographs and lateral cephalometric tomograms were assessed at the following intervals: before surgery (T1), immediately after surgery (T2), 6 to 12 months after surgery (T3), and at the longest follow-up (T4). Lateral cephalometric tracings were superimposed to calculate surgical change (T2 - T1), short-term stability (T3 - T2), and long term stability (T4 - T3) of the orthognathic surgery procedures. Maximum interincisal opening (MIO) and subjective TMJ pain (visual analog scales) were comparatively evaluated at T1 and T4. RESULTS: Subjective TMJ pain levels and MIO improved in all 3 groups after surgery. Before surgery, 56 of 70 patients (80%) had pain and 14 of 70 patients (20%) had no pain. At the longest follow-up, 42 of 70 patients (60%) reported complete relief of TMJ pain. Only 5 of 70 patients (7%) had severe pain after surgery compared with 37 of 70 patients (53%) before surgery. At the longest follow-up, 6 of 70 patients (9%) showed less than 35 mm MIO, residual severe pain, or both. One patient had significant condylar resorption after surgery. The orthognathic procedures were found to be stable in the long term. Concomitant TMJ and orthognathic surgery had an overall success rate of 91.4% based on a greater than 35 mm MIO and a decrease in pain. CONCLUSIONS: When indicated, TMJ and orthognathic surgery can be concomitantly performed with predictable results and a good success rate. Strong consideration should be given to early surgical intervention because the success rate decreases significantly with pre-existing TMJ dysfunction of greater than 48 months' duration. PMID- 11928089 TI - Acoustic and perceptual analysis of the sibilant sound /s/ before and after orthognathic surgery. AB - PURPOSE: Orthognathic surgery may have a positive or negative effect on speech. Perceptual evaluation of presurgical and postsurgical articulation is difficult because speech errors, when they occur, are usually fricative distortions, which may be difficult to document reliably. In this study, acoustic analysis was used to supplement perceptual judgment of presurgical and postsurgical productions of /s/. SUBJECTS AND METHODS: The study population consisted of 9 Cantonese speakers undergoing osteotomy for Class III skeletal deformity and 9 age- and gender matched adults with normal occlusion and speech. The speech sample consisted of 6 words with the initial sibilant sound /s/. Perceptual analysis included narrow phonetic transcription and classification of error types. Acoustic analysis included measurement of first and second spectral peaks, fricative duration, noise bandwidth, and noise-to-vowel decibel ratio. RESULTS: The results of the perceptual analysis showed a decrease in articulatory errors for the group after surgery, although 5 patients had no perceptual errors before surgery. Acoustic analysis showed significant differences between the experimental and control groups before surgery for 2 variables (spectral peak I and bandwidth). Three months after surgery there were no significant differences between the control group and the experimental group, except for bandwidth. Twelve months after surgery, there were significant differences between the 2 groups in noise bandwidth and spectral peak II. CONCLUSIONS: The results suggest a possible relapse at 1 year after surgery, based on spectral peak values. Osteotomy appears to result in a positive change in articulation for most patients, but speech outcome after osteotomy must be evaluated both 1 year and shortly after surgery. PMID- 11928091 TI - Reduction in the incidence of alveolar osteitis in patients treated with the SaliCept patch, containing Acemannan hydrogel. AB - PURPOSE: In the present study, we compared the incidence of alveolar osteitis (AO) in patients treated with either clindamycin-soaked Gelfoam (Pharmacia and Upjohn Co, Kalamazoo, MI) or SaliCept Patches (Carrington Laboratories, Inc, Irving, TX). The SaliCept Patch is a freeze-dried pledget that contains Acemannan Hydrogel (Carrington Laboratories) obtained from the clear inner gel of Aloe vera L. PATIENTS AND METHODS: A retrospective evaluation was performed of the records of 587 patients (1,031 sockets) whose extraction sites had been treated with clindamycin-soaked Gelfoam. A prospective trial was conducted in which 607 patients (1,064 sockets) had 2 SaliCept Patches placed immediately after extraction. The same surgeon treated all patients. RESULTS: Analysis restricted to mandibular third molar sites showed that 78 of 975 sites (8.0%) in the Gelfoam group developed AO, whereas only 11 of 958 sites (1.1%) in the SaliCept group developed AO (P <.0001). Further analysis of all extraction sites revealed that the incidence of AO in the Gelfoam group was 7.6% compared with 1.1% in the SaliCept-treated group (P <.0001). CONCLUSIONS: The study results suggest that the SaliCept Patch significantly reduces the incidence of AO compared with clindamycin-soaked Gelfoam. PMID- 11928093 TI - Follow-up in patients with oral cancer. AB - PURPOSE: To provide standardized follow-up of patients who underwent oral cancer treatment, the German-Austrian-Swiss Cooperative Group on Tumors of the Maxillofacial Region (DOSAK) established a schedule of check-up examinations for a 5-year period. MATERIALS AND METHODS: On the basis of a questionnaire, we investigated oncologic follow-up in respect to early detection of recurrences, cost and outcome efficiencies, and physician and patient judgment in the development of impairments. RESULTS: Only 11% of all operated patients participated regularly in the follow-up program within the first 3 years. Ultrasound was the most effective imaging for the detection of recurrences. Swallowing, speaking, tongue mobility, and facial appearance were the most common problems. Troubles with swallowing were mainly induced by poorly fitted dentures, discontinuity of the mandible, osteomyelitis, and xerostomia. Outer appearance was mostly impaired by scars, missing facial muscle function, and edema. The problem of chronic pain remained unsolved. CONCLUSION: Due to the sequelae of treatment, this study shows the need for close medical and psychological follow up. PMID- 11928095 TI - Dentoalveolar distraction osteogenesis for rapid orthodontic canine retraction. AB - PURPOSE: We present a technique to reduce the overall orthodontic treatment time by means of dentoalveolar distraction osteogenesis. PATIENTS AND METHODS: Eleven patients who were planned to undergo orthodontic treatment with bilateral first premolar extractions and subsequent bilateral canine tooth distalization underwent osteotomy around the canine tooth. The first premolar was extracted, and the buccal bone was carefully removed. After wound closure, a special orthopedic device was mounted and cemented to the first molar and canine teeth. Distraction was started the same day at the rate of 0.4 mm twice a day and continued until adequate movement of the canine teeth was achieved. The device was then removed, and orthodontic therapy was continued with fixed appliances. RESULTS: The distraction rate and the device were well tolerated by all of patients. No anchorage loss in the second premolar and first molar teeth, root resorption, dental ankylosis, discoloration, or loss of vitality was detected. CONCLUSION: The concept of distraction osteogenesis for rapid orthodontic tooth movement is promising and feasible for clinical practice. PMID- 11928096 TI - Transverse displacement of the proximal segment after bilateral sagittal osteotomy. AB - PURPOSE: The aim of the present investigation was to evaluate the transverse displacement of the proximal segment and ramus rotation after a bilateral sagittal osteotomy (BSO) with rigid internal fixation (RIF) using bicortical LAG screws. PATIENTS AND METHODS: We conducted a retrospective review of 37 patients (14 males and 23 females, age range of 14 to 55 years) who underwent a mandibular advancement with BSO and RIF. Posteroanterior and lateral cephalometric radiographs were obtained 1 to 8 weeks before and 1 to 4 weeks after surgery. The transverse displacement and angulation of the proximal segments after surgery were measured on posteroanterior radiographs, using the best-fit method. The amount of mandibular advancement was compared with the amount of transverse displacement of the proximal segments. RESULTS: In the 1 to 4-week postoperative period after a BSO, 36 of 37 subjects showed an increased transverse intergonion distance (5.6 mm) (P <.0001) and 35 of 37 patients showed an increased transverse interramus width (3.3 mm) (P <.0001). No correlation was found between mandibular advancement and transverse displacement of the proximal segment. CONCLUSIONS: The study results indicate that transverse displacements of the proximal segments occur with BSO and RIF. The clinical impact on temporomandibular joint symptomatology or surgical relapse with such displacement was not assessed in the study. Future studies that address these issues may help to determine whether there is an association between proximal segment displacement and surgical relapse, temporomandibular dysfunction, or both. PMID- 11928097 TI - Speech after partial glossectomy: a comparison between reconstruction and nonreconstruction patients. AB - PURPOSE: This study was undertaken to compare the articulation intelligibility of patients after hemiglossectomy or minor glossectomy within the hemitongue with or without reconstruction. MATERIALS AND METHODS: The articulation intelligibility of 19 patients who underwent hemiglossetomy or minor glossectomy within the hemitongue was investigated at least 6 months after surgery with an articulation intelligibility test with 40 sensitive Chinese sounds. Patients were divided into group 1 (those undergoing reconstructive surgery after glossectomy) and group 2 (those not undergoing reconstructive surgery after glossectomy). Defects of the tongue (in group 1) were reconstructed immediately with free radial forearm flaps or pedicled flaps after glossectomy and closed primarily without reconstruction in group 2. The articulation intelligibility scores were compared between the 2 groups. RESULTS: The mean articulation intelligibility score for group 1 was 77.0% (blade portion glossal sounds), 76.3% (mid portion glossal sounds), and 84.7% (rear portion glossal sounds), and those of group 2 were 94.6%, 92.1%, and 95.3%, respectively. The intelligibility of articulation of group 2 was significantly higher than that of group 1 in blade portion glossal sounds (P <.05), mid portion glossal sounds (P <.05), and rear portion glossal sounds (P <.01). CONCLUSION: The articulation intelligibility was better in patients not receiving grafts than in those receiving grafts. Reconstruction with flaps, which may interfere with the flexibility and mobility of the tongue, may contribute to articulatory impairment. If speech is the outcome of interest, reconstruction may be unnecessary with hemiglossectomy or other partial (minor) glossectomy within the hemitongue. PMID- 11928098 TI - Metastasis of maxillary carcinoma to the parapharyngeal space: rationale and technique for concomitant en bloc parapharyngeal dissection. AB - PURPOSE: En bloc resection of the primary tumor and regional lymph nodes is the classic method of surgery in cases of head and neck cancer, but it is not performed in cases of carcinoma of the maxillary gingiva or antrum for anatomic reasons. One of the reasons for the poor prognosis of patients with maxillary cancer and N+ stage necks is thought to be recurrence in the parapharyngeal space, which is out of the surgical field in radical neck dissection. The purpose of this study was to discuss the rationale and indication for en bloc resection and parapharyngeal dissection for maxillary cancer. PATIENTS AND METHODS: Ninety nine patients with maxillary cancer (54 in the gingiva and 45 in the antrum) treated at our institution between 1980 and 2000 were studied retrospectively. RESULTS: In 4 patients, there was recurrence in the parapharyngeal spaces despite good control of tumors in the maxilla and the neck. These 4 patients had all undergone resection of maxilla and neck lymph nodes separately. We also report the case of a patient with carcinoma of the maxillary antrum who underwent en bloc resection of the maxilla and neck. After radical neck dissection, parapharyngeal dissection was performed with a mandibular ramus osteotomy approach, and the maxilla and neck tissue were resected en bloc. CONCLUSIONS: Although en bloc resection causes more extensive surgical damage, it may be useful in patients with maxillary cancer who have metastasis in the upper jugular lymph nodes. PMID- 11928100 TI - Determination of subepithelial connective tissue graft thickness in the dog. AB - PURPOSE: In this study, we determined the tissue thickness of healed subepithelial connective tissue grafts, the type of tissue present after healing, and the percentage of shrinkage of subepithelial connective tissue grafts over time in the facial aspect of the canine maxilla. MATERIALS AND METHODS: Four dogs each received 2 single-thickness subepithelial connective tissue grafts, 2 double thickness grafts, and 2 sham-operated sites in the facial aspect of the maxilla bilaterally. The subepithelial connective tissue graft was harvested from the palate and included the periosteum. After the creation of a pouch on the facial aspect of the maxilla, the grafts were sutured directly to the maxillary periosteum and additional sutures were placed to secure the overlying mucosal flap to the graft. Thickness measures of the grafts before placement and after surgery with the grafts in place were made biweekly with a calibrated probe using known reference points on the teeth. Thickness measures were analyzed using a random-effects mixed model. All dogs were killed at 12 weeks, and the tissues were fixed, serially sectioned, and stained with hematoxylin and eosin for microscopic examination. RESULTS: Initial measurements of the sites demonstrated a thickness of 1.5 +/- 0.6 mm. Immediate postoperative measures were 7.1 mm (single thickness), 7.0 mm (double thickness), and 4.8 mm (sham operated). Week 6 measures were 3.9 mm (single thickness), 3.6 mm (double thickness), and 2.7 mm (sham operated), and week 12 measures were 3.9 mm (single thickness), 3.4 mm (double thickness), and 2.9 mm (sham operated). The difference between single- and double-thickness grafts was not significant (P =.34). The differences between single thickness and sham operated (P =.0036) and between double thickness and sham operated (P =.0049) were significant. Microscopic evaluation demonstrated that the grafts were indistinguishable from the adjacent subcutaneous tissues of the maxilla. The submucosal graft region contained fat, small blood vessels, nerves, and fibrous connective tissue. CONCLUSIONS: Based on this study in dogs, shrinkage of subepithelial grafts can be expected. The grafts become incorporate into the host tissues and appear similar to the normal connective tissue. PMID- 11928101 TI - Effect of calcium phosphate surface coating on bone ingrowth onto porous-surfaced titanium alloy implants in rabbit tibiae. AB - PURPOSE: The purpose of the present study was to determine whether calcium phosphate coating has a significant impact on bone ingrowth into a porous titanium implant. MATERIALS AND METHODS: Porous-surfaced titanium alloy Ti-6Al-4V implants were prepared with or without the addition of a thin surface layer of calcium phosphate applied by sol-gel coating. Implants were placed into the tibiae of 16 rabbits. Implanted sites were allowed to heal for 2 weeks, after which specimens were retrieved for morphometric assessment using backscatter scanning electron microscopy. RESULTS: The data collected show that there is more extensive ingrowth into the porous regions of the calcium phosphate-coated implants than into the control implants. The weighted average ingrowth for the calcium phosphate-coated implants was 2.01, whereas that for the noncoated implants was 1.49; the difference is statistically significant (P <.01). CONCLUSIONS: The addition of a thin layer of calcium phosphate to these implants appears to promote a more extensive implant-to-bone interface by allowing the neck regions to become intimately ingrown with bone even after only 2 weeks of initial healing. PMID- 11928103 TI - Effects of fixation type and guided tissue regeneration on maxillary osteotomy healing in rabbits. AB - PURPOSE: The development of fibrous nonunions after orthognathic surgery is thought to result from an interaction of biomechanical stress and the differential and more rapid migration of fibroblasts (compared with osteoblasts) into the wound site during healing. The present study was designed to test this hypothesis through the manipulation of guided tissue regeneration and osteotomy fixation techniques in an experimental rabbit model. MATERIALS AND METHODS: Bilateral critical size (4 mm) defects (n = 24) were produced in the maxillae of 12 adult New Zealand white rabbits. The maxillary segments were rigidly or nonrigidly fixed using bone microplates and screws or osteosynthetic wires. The defects were then covered with a resorbable collagen membrane or left uncovered. The rabbits were followed for 4 weeks with the use of serial dorsoventral and lateral oblique cephalographs, and the maxillae were then harvested for histologic analyses. RESULTS: Radiographic and histomorphometric analyses revealed that rigidly fixed defects, covered with membrane, showed the most rapid and organized new bone formation. The rigidly fixed defects with the membrane averaged approximately 40% more new bone in the osteotomy site compared with the rigidly fixed defects with no membrane. Nonrigidly fixed defects with no membrane showed an ingrowth of fibroblasts and fibrous nonunions. CONCLUSIONS: These experimental results suggest that an interaction between the decreased fibrous tissue ingrowth through guided tissue regeneration and osteotomy segment stability from rigid fixation prevented postoperative fibrous nonunions and facilitated new bone regeneration and osteotomy site healing in this rabbit model. PMID- 11928105 TI - Multiple pigmented lesions of the lower lip. PMID- 11928106 TI - Management of acute suppurative parotitis. PMID- 11928107 TI - Deep lobe parotid gland lipoma. PMID- 11928108 TI - A case of benign fibrous histiocytoma of the upper lip in a 6-month-old infant. PMID- 11928109 TI - Maxillofacial hydatid cyst. PMID- 11928110 TI - Inferior alveolar nerve paresthesia associated with a mandibular dentigerous cyst. PMID- 11928111 TI - Neonatal bilateral idiopathic hyperplasia of the coronoid processes: case report. PMID- 11928112 TI - Parotitis caused by Salmonella choleraesuis varietas Kunzendorf. PMID- 11928113 TI - External auditory canal carcinoma involving the temporomandibular joint: two cases presenting as temporomandibular disorders. PMID- 11928114 TI - The orbicularis oculi muscle flap in subciliary access for management of orbital trauma: Technical note. PMID- 11928115 TI - Oral endotracheal intubation: median submental (retrogenial) approach. PMID- 11928116 TI - Lingual traumatic ulceration (Riga-Fede disease). PMID- 11928117 TI - [Coloration of cytologic thick sections containing biomaterials with silver methenamine. Usefulness for scanning electron microscopy]. AB - Prostheses implanted in hard tissues cannot be processed for electron microscopic examination or microanalysis in the same way as those in other tissues. For these reasons, we have developed a method allowing light and electron microscopic studies as well as microanalysis of the interface between bone and biomaterials after the sections glycoproteins have been stained with silver methenamine. Silver can be evidenced by SEM in back scattered mode. MATERIALS AND METHODS: Tranverse sections of a human femur containing an HA-coated prostheses were obtained with a diamond saw and ground to a thickness of 50-100 microns. The sections were stained in a microwave oven using a 1% silver methenamine solution. They have been examined by back-scatter SEM operated at 25 KV. EDS has been performed on cellular inclusions and extracellular bone matrix. RESULTS: Type I and III collagen fibers, and reticulin fibers were stained. The mineralized matrix was heavily colored. At the cell level, the nuclear and cytoplasm membranes, the chromatin and ribosomes were shown. The characteristic peaks of the Ag spectrum are distinct from those of the elements used in orthopaedic biomaterials and did not impair their identification. PMID- 11928118 TI - Urethral cytology and diagnosis of the pregnancy term. About some observations on the use of this method in an hospital setting. AB - A retrospective study of routine urethrocytograms performed over a 10-year period in 3,478 pregnant women for the diagnosis of pregnancy term was carried out to determine the reliability of urethral cytology when two, three, or four successive urethral smears were performed at 2- or 3-day intervals, and to investigate the correlations between the cytological results and the neurological ages of newborns. Microscopical examination of the 3,478 routine smears demonstrated the existence of a before-term aspect, a near-term aspect, or an at term aspect in 7.4%, 66.7%, and 22.1% of cases, respectively. The diagnosis of post-term was noted only in 1 smear. Repeated urethrocytograms (1, 2, or 3 smears after the firstone) in women with a previously diagnosed before-term aspect or a near-term aspect revealed the occurence of the at-term aspect over time. Concordant correlations between cytological results and the neurological ages of the newborns were noted in 98.1% of cases. Three developmental stages (beginning, middle, end) of the near-term aspect and three stages of the at-term aspect were described. Due to its interest, the urethral-smear method should be worth generalizing as the other laboratory examinations that are routinely used to determine the term of pregnancy. PMID- 11928119 TI - [Disappearance of the avian teeth or "when birds had teeth"]. AB - Authors describe the evolution of the avian lineage since its origin to our days and in particular the characteristics of the Jurassic or Cretaceous periods. They evoke the mechanisms which explain the involution of birds' teeth, their complete disappearance, birds nowadays having a beak instead. They stress the importance of natural selection and heterochrony. PMID- 11928120 TI - [Osseointegration of 2 different types of calcium phosphate materials: ceramics and ionic cements]. AB - Different kinds of calcium phosphate biomaterials can be used as bone substitutes. Ceramics are constituted by HA or TCP grains linked by grain boundaries. Their porosity depends on the powder characteristics and the sintering temperature. It can be very low with a pore size inferior to one micron. The setting of calcium phosphate hydraulic cements results from the precipitation of a calcium phosphate phase different from the one in suspension in the paste. The strength of the cement is given by the entanglement of the growing mineral crystals. Calcium phosphate hydraulic cements and ceramics have very different physico-chemical characteristics. We have studied the histological integration of both kinds of material. The first material was constituted by macroporous ceramics composed of 75% HA and 25% beta-TCP, the cement was made of beta-TCP grains dispersed in a DCPD matrix. The sequence of events which leads to the ceramic integration is always the same: a/ ingrowth of a loose connective tissue; b/ osteoblast differentiation from fibroblast-like cells of the connective tissue in close proximity to the implant surface; c/ osteoid synthesis at the ceramic surface toward the pore center; d/ remodeling of the immature bone and the ceramic itself. The cement is differently integrated. The osteoblasts differentiate at some distance from the implant and there is a trabeculae ingrowth toward the material. CONCLUSIONS: The early stages of both materials osteointegration are different. The integration is centrifugal for ceramics and centripetal for the cement. PMID- 11928121 TI - [Tartrate-resistant acid phosphatases (TRAP) and cytology of pseudosynovial membranes]. AB - Pseudo-synovial membranes are found at the interface between bone and aseptically lost prostheses. They are supposed to be due to the release of material debris at the interface with bone. They are mostly composed of macrophages and giant cells, and some fibrocytes in the deep layer of the membrane. The different histological structures present in these membranes have been largely described. Very recently, Kadoya et al. have demonstrated that a lot of giant cells present in the pseudo synovial membranes show a tartrate resistant acid phosphatase activity in the presence of more than 100 mM tartrate. They have also shown that some of these cells were responsible for bone degradation. We have studied the tartrate acid phosphatase activity of a series of 10 membranes containing various material debris: titanium alloy, stainless steel, polymethylmethacrylate, polyethylene. Many mono or multinucleated cells showed a TRAP activity in their lysosomes in the presence of a 400 mM tartrate concentration. In certain membranes, these cells showed a TRAP activity located in the nucleus. This study shows that the macrophage population is very heterogeneous and the difference between osteoclasts and giant cells may be unclear. PMID- 11928122 TI - Ultrastructure of the atrioventricular junctional area in the heart of Molossus molossus Pallas 1766 (Chiroptera: Molossidae). AB - The atrioventricular junctional area (AVJA) consists of a group of structures that connects the atrial and ventricular myocardium. Five hearts of an insect eating bat were studied in light and transmission electron microscopy. In M. molossus, the AVJA consists in a mass of muscle fibers intermingled with variable amount of connective tissue and blood vessels surrounded by the adjacent myocardium and the attachment of the right atrioventricular and aortic valves in the fibrous skeleton. In light microscopy, conducting cells of the AV node and bundle can be distinguished from working cells: smaller size, paler staining reaction and the presence of e sheath of connective tissue surrounding each cell (largely composition by type I collagen fibers). Three cell types are observed in the AVJA. Nodal cells are irregular with few cytoplasmic organelles and several slender sarcolemmal modifications. Myofibrils are sparse and not clearly observable. Transitional cells are spindle-shaped and grouped together into bundles. The cytoplasm, poor in glycogen, has scarce electron-density and myofibrils organized into sarcomeres. Caveolae is observed randomly distributed at the periphery of the cell. The AV bundle cells are elongated with clusters of myofibrils organized in the periphery and a glycogen free area around the nucleus. Ventricular cells are bigger than the atrial ones and show well developed myofibrils in alternated rows with mitochondria. Lipid droplets are seen near mitochondria and glycogen granules. Intercalated discs and T-tubules are found in working cells but not in conducting ones. The fibrous skeleton has collagen fibers intercalated with fibroblasts. PMID- 11928123 TI - [Brief communication. Study of a supernumerary neck muscle: atlas rhomboid muscle (Macalister, 1871)]. AB - We observed a muscle originating from the lateral process of Cl and inserting on the anterior aponeurosis of the rhomboideus major muscle. This accessory muscle was present on the right side only. PMID- 11928124 TI - Polymerase Chain reaction generated probes for fluorescence in situ hybridization. AB - The extended use of Fish with centromeric probes in many cytogenetic laboratories is often impaired by the cost of this technique. Polymerase Chain Reaction (PCR) constitutes a simple way to generate and label such centromeric probes at low cost. Two types of human DNA source can be used: 1--Somatic hybrid cell lines containing a unique human chromosome. The specific amplification of the human subset of alphoid DNA is realised with a primer pair specific for the consensus region of human alpha satellite sequence. 2--Total Human DNA. This time, a primer pair specific for the alpha satellite DNA of the chromosome of interest must be designed. These probes, labelled during the PCR reaction by direct incorporation of modified dUTP, are actually widely used in our laboratory, alone or mixed with other probes (chromosome painting or locus specific probes). PMID- 11928125 TI - A unique anomaly of the fibrous sling of the digastric muscle. AB - Anomalies of digastric muscle are not very rare. During a routine dissection, it was found on a cadaver that the digastric muscles on both sides had no fibrous slings. It was also observed that the two intermediate tendons crossed over the stylohyoid muscles. Such anomalies should be considered during the evaluation of the floor of the mouth after CT examination of MR imaging, and during surgical procedures as well. PMID- 11928126 TI - Morphometric analysis of fibroblasts of the mammary lobular stroma during the follicular and luteal phases of the menstrual cycle. AB - We determined the nuclear volume of fibroblasts of the normal mammary lobular stroma during the follicular and luteal phases of the menstrual cycle. Twenty patients aged 15 to 35 years and eumenorrheic for at least 6 months were randomly assigned to 2 groups, i.e., 10 women in the follicular phase and 10 in the luteal phase. The nuclear volume was 34.4 micron 3 and 98.8 micron 3 for the follicular and luteal phases, respectively, with the difference being statistically significant (p < 0.05). These data suggest a higher metabolic activity in the mammary intralobular stroma during the luteal phase of the menstrual cycle, probably due to a synergistic action of estradiol and progesterone. PMID- 11928127 TI - Off the shelves and into the streets. PMID- 11928128 TI - Secondary analysis: theoretical, methodological, and practical considerations. AB - Secondary analysis, which involves the use of existing data sets to answer new research questions, is an increasingly popular methodological choice among researchers who wish to investigate particular research questions but lack the resources to undertake primary data collections. Much time loss and considerable frustration may result, however, if researchers begin secondary analyses without an awareness of the distinctive methodological and practical challenges involved. This article highlights difficulties that may arise when researchers use data from previous clinical research projects, including theoretical issues and problems involving sampling, measurement, and external and ecological validity. It also offers practical suggestions for undertaking a secondary analysis and criteria for evaluating secondary analyses. PMID- 11928129 TI - "Are we in Kansas yet, Toto?" The construction of chronic illness in research. PMID- 11928130 TI - Parents' perceptions of chronic illness trajectories. AB - The notion of a small, generic set of chronic illness trajectories that can be independent of specific medical diagnoses, though controversial, has some theoretical, clinical, and qualitative research support. The purpose of this study was to quantitatively describe trajectories among parents of children with a chronic condition. It was hypothesized that factor analysis would confirm 3 trajectories similar to those in the qualitative literature and that parents' perceptions of their child's trajectory would differ significantly from medically based perceptions. A total of 140 parents provided data on their perceptions of the past, present, and future course of the condition of their repeatedly hospitalized child. Fourteen time-related items from the Coping Health Inventory for Parents Questionnaire on Resources and Stress and the Parenting Stress Index were analyzed. Pre- and post-hospitalization factor analyses extracted the same 8 items to construct 3 trajectories: Life Threatening; Declining; and Stable, Optimistic. The views of approximately one third of the parents differed from medically based classifications. Type of nursing care had no bearing on the perceptions of the parents. PMID- 11928131 TI - Shortage of nurses, shortage of nursing. PMID- 11928132 TI - Living with chronic illness: the interface of stigma and normalization. AB - Traditionally, researchers have studied and interpreted the chronic illness experience through a lens of either stigma or normalization, but rarely both simultaneously. When chronic illness is examined through a stigma lens, the findings tend to focus on the manner in which the individual suffers from the stigma. When it is examined through a normalization lens, the findings tend to articulate the ways in which the individual achieves normalcy despite having a chronic condition. This paper discusses the implications of assuming either of the two perspectives independent of the other. The authors argue that, in order to capture and understand the dynamic and evolving experience of people with chronic conditions, researchers should consider the interdependence of the two perspectives and avoid assumptions that derive from stigma or normalization alone. Considering stigma and normalization aspects of a chronic illness experience, in interaction over time, will facilitate a broader and more accurate understanding of the complex experience of people coping with chronic conditions. PMID- 11928134 TI - Schizophrenia and life in the world of others. AB - This article examines the social and relational impact of chronic schizophrenia on 10 adult sufferers living in the community in New Zealand. The findings reported here are drawn from a larger Heideggerian phenomenological study that set out to answer the question What is it like to live with schizophrenia? While schizophrenia affects all dimensions of a person's life, in this article 6 themes are presented to illustrate the impact of their illness on one aspect of the participants' lives--their interactions with others. The themes are: living with the prejudice of others, being fearful of others, feeling uncomfortable in the company of others, staying engaged with others in the world, depending on others for help, and finding others who understand. The article demonstrates that it is possible to understand the experience of those with schizophrenia, and suggests that attention to the relational aspects of the lives of people with this illness can help them to lead healthier and happier lives. PMID- 11928133 TI - Trajectory of certain death at an unknown time: children with neurodegenerative life-threatening illnesses. AB - Childred with neurodegenerative life-threatening illnesses (NLTIs) account for a significant proportion of children requiring palliative care. Most of their care is provided at home by their families over many years, yet there is a paucity of research examining families' experiences with a child with an NLTI is dying at home. In this grounded theory study, data were collected from 8 families through observations and audiotaped interviews. Families moved through a process of navigating uncharted territory as they lived with their dying child. The illness trajectory of cetain death at an unknown time was not a steady decline. Instead, families lived much of their lives on plateaus of relative stability where they often felt alone and isolated from health-care professionals. Inevitable, periods of instability originated in subsequent precipitating events in the process that led to families dropping off the plateau on the way to the child's inevitable death. Implications for research and practice are discussed. PMID- 11928135 TI - Chronic illness in the next millennium: context and complexity. PMID- 11928136 TI - Physical activity as a mediator between dyspnea and fatigue in patients with chronic obstructive pulmonary disease. AB - A cross-sectional design was used to examine the mediating effect of physical activity between dyspnea and fatigue in patients with chronic obstructive pulmonary disease (COPD). Dyspnea was measured using a vertical visual analogue scale, fatigue using the Profile of Mood States-Fatigue subscale, and physical activity using the 6-minute-walk (6MW) test. A convenience sample of 17 male and 22 female patients with COPD provided data for analysis. The sample was characterized by relatively high forced expiratory volume in 1 second (FEV1) of predicted normal values, indicating mild lung impairment and high mean levels of fatigue and dyspnea. Controlling for age and FEV1, dyspnea, physical activity, and fatigue were significantly interrelated. Results of the regression analysis suggested the mediating function of physical activity between dyspnea and fatigue. Limitations and suggestions for practice and future research are presented. PMID- 11928137 TI - Capturing day-to-day aspects of living with chronic illness: the need for longitudinal designs. PMID- 11928138 TI - [Syndromes and sequences]. AB - In order to introduce a series of articles on syndromes, the differences between syndromes and diseases are discussed, their causes summarized, annotations are made with regard to genotype and phenotype, and the diagnosis and frequency are described. PMID- 11928139 TI - [Syndromes. 1. Cleidocranial dysplasia]. AB - Dysostosis cleido-cranialis is mainly characterized by a pathognomonic deformity of the skull, a hypoplastic midface, lack of eruption of permanent teeth, supernumerary teeth, defects in the clavicles, short stature and multiple other skeletal deformities. The inheritance is autosomal dominant, but there is a considerable number of new mutations. PMID- 11928140 TI - [Long-term results of traditional periodontal therapy]. AB - OBJECTIVE: To evaluate the long-term results of traditional therapy consisting of initial preparation followed by mucogingival surgery including bone recontouring and regular maintenance treatments. DESIGN: 125 patients between 25 and 70 with a generalised periodontitis of the type adult and rapidly progressing periodontitis were selected. These patients were treated according to a fixed protocol by a periodontist and a dental hygienist. SETTING: The Practice for Periodontology in Groningen. METHODS: Patients were treated in a practice limited to periodontology. Before and 5 years after treatment the following data were assessed: plaque- and bleeding-score, probing pocket-depth, mobility and furcation involvement. Furthermore the reason for extraction was established. RESULTS: In general a great reduction in plaque- and bleeding-score and probing pocket-depth was much better in case of good oral hygiene. Within 5 years after treatment only 1.1% of the teeth were extracted. Furcations with through and through involvement in combination with deep residual pockets were mostly the reason for this. CONCLUSION: It is concluded that with traditional periodontal therapy following a strict protocol including regular maintenance treatments excellent treatment results can be achieved. PMID- 11928141 TI - [The use of a thyroid collar for intraoral radiography]. AB - OBJECTIVE: To determine whether a thyroid collar is a reasonable measure to reduce patient exposure from intraoral radiography (cost benefit analysis). DESIGN: In the thyroid gland of a Rando phantom dose measurements were carried out to determine the effect of a thyroid collar during intraoral radiography. SETTING: Department of Oral Radiology at ACTA, Amsterdam. METHODS: Dose measurements were carried out using LTDs. The average absorbed dose to the thyroid gland with and without thyroid collar from intraoral radiography was compared using an analysis of variance. RESULTS: For periapical radiographs the equivalent dose to the thyroid gland was significantly lower (p < 0.05) when a thyroid collar was used. For bitewing radiography there were no significant effects of the thyroid collar (p > 0.05). The cost benefit analysis showed that it takes more than 40 years before the benefits of a thyroid collar exceed the costs. CONCLUSION: Collective use of thyroid collars therefore does not seem to be a reasonable measure to optimize radiological protection during intraoral radiography. PMID- 11928142 TI - [A crooked face]. PMID- 11928143 TI - [Tuberculosis 1]. PMID- 11928144 TI - [Layer thickness of dental adhesives. Relation between layer thickness and viscosity]. AB - OBJECTIVE: To investigate the viscosity of two dental adhesives in relation to the adhesive layer thickness. DESIGN: In vitro study. METHODS: The viscosity of several dental adhesives has been determined with a reogoneometer. Both a high and a low viscous adhesive was used to restore class II preparations. These restorations were sectioned and the thickness of the adhesive layer was measured on several defined locations. A statistical analysis was performed. RESULTS: The high viscous adhesive resulted in a thinner layer along the cervical outline of the restoration. With both materials pooling occurred at the line angles of the preparation. On the flat surfaces of the preparations there was a layer of even thickness. CONCLUSION: The differences in thickness of the adhesive layer as often seen on radiographs cannot be explained solely by the use of adhesives with a different viscosity. PMID- 11928145 TI - [Complications in the treatment of angle class II division 1 malocclusion]. AB - OBJECTIVE: This article describes a retrospective inventory of various categories of complications which occurred in a sample consecutively treated patients with Class II division 1 malocclusion. DESIGN: Retrospective, descriptive study. METHODS: Treatment was performed with a headgear-activator and a edgewise appliance. All patients originate from one orthodontic specialist office and were between 10 and 14 years of age during treatment. From the group of 600 patients 100 (n = 100) were randomly selected for the study. A standard form was used for the inventory of the complications. The complications were divided into 5 categories: dental, skeletal, functional, psychosocial and multiple (two or more complications). RESULTS: In the group of 100 patients no complications were found in 48 patients (48%); 52 patients showed complications (52%). From these 52 patients 29 showed one complication, 19 had 2 complications, 4 showed 3 complications. In total 79 complications occurred divided as follows: dental complications 23 (29.1%); skeletal complications 19 (24.1%); functional complications 15 (19.0%); psychosocial complications 22 (27.8%). CONCLUSION: Complications occur very frequently in the treatment of Angle Class II division 1 malocclusions. These complications may occur in various combinations and may compromise the treatment result in a variable way. PMID- 11928146 TI - [Condylar hyperplasia. Early detection is desired]. AB - When single-sided excessive condylar growth is detected in time, removal of the affected condyle is sufficient. In cases of late detection, extensive surgery of upper and lower jaw and the chin may be necessary. PMID- 11928148 TI - [Syndromes 6. Crouzon's disease]. AB - Morbus Crouzon is a syndrome with craniosynostosis. Besides a deviating form of the skull this causes a retropositioned middle third of the face. Therefore pseudo-exophthalmos, strabismus and an Angle Class III relation of the jaws are usually seen. The eyes are especially vulnerable. Since the symptoms are rather symmetrical, a good result of the corrections can usually be achieved. The form of the skull will be corrected in very early childhood, while the deformities of the face are by preference operated after completion of growth. However, different reasons can be found to start correction in an earlier stage. PMID- 11928147 TI - [Comparison of dental implant systems for mandibular overdentures]. AB - OBJECTIVE: To compare three implant systems in patients with severely resorbed mandibles 1 year after insertion of the mandibular denture. DESIGN: A prospective multicenter randomized clinical trial. SETTING: Centers at University of Nijmegen and Groningen. METHODS: Three different implant systems were used: the transmandibular implant, the IMZ and the Branemark system. Treatment was assigned to 88 patients according to a balanced allocation method. Evaluation included clinical and radiographic parameters. A Clinical Implant Performance scale (CIP) was constructed based on all conceivable complications of the different implant systems. RESULTS: During the healing period 1 IMZ- and 1 BRA-implant were lost. One TMI was removed after functional loading. The results of the clinical and radiographic parameters and the CIP-scale showed no significant differences between the three implant systems. CONCLUSION: Taking the one year evaluation into account the three systems used did not differ in clinical and radiographic performance. PMID- 11928149 TI - [An innocent swelling of the upper lip?]. PMID- 11928150 TI - [Cervix dysplasia and cervix carcinoma]. PMID- 11928151 TI - Nursing's valued resources: critical issues in economics and nursing care. PMID- 11928152 TI - Costs of postpartum care: examining associations from the Ontario mother and infant survey. AB - A cross-sectional survey of 1,250 mothers of "normal" newborn infants was conducted to assess mother and infant characteristics as well as the costs of health and social services used in the first 4 weeks after discharge from hospital in the province of Ontario, Canada. Each mother was asked to recall use of services for herself and her infant. This utilization was multiplied by the unit cost of each service and summed to arrive at the total cost of services used. Although re-admission rates were low (1% for mothers, 4% for infants), costs associated with hospital and emergency room care ($194 on average per mother/infant dyad) accounted for the greatest proportion of total health-care costs. Physician or midwife visits, which were received by almost all mothers, were the next most costly service ($128 on average per dyad). Mothers with incomes under $20,000 had more medical costs than those with higher incomes. Costs of community nursing care ($86 on average per dyad) were higher for mothers with more than 5 self-identified learning needs (e.g., infant feeding, infant care and behaviour, emotional changes in self, signs of illness in infant); perception of their own health as poor; perception of inadequate help and support at home; many signs and symptoms of depression; and postpartum hospital stay of 48 hours or less. Costs for social work visits were higher for mothers with depression and mothers with low incomes. Total health and social service costs were double for mothers with family incomes under $20,000 ($788 vs. $399 on average per dyad) and for mothers with clinical depression ($845 vs. $413). Specifically, nursing care costs were greater for mothers with high depression scores ($135 vs. $81). PMID- 11928155 TI - Willingness-to-pay (WTP): the new-old kid on the economic evaluation block. PMID- 11928156 TI - The costs and effects of addressing the needs of vulnerable populations: results of 10 years of research. PMID- 11928154 TI - Towards ethical inquiry in the economic evaluation of nursing practice. AB - Economic evaluation is a critical tool for nursing and health care. The authors claim that economic inquiry needs to be supported by expertise in ethical inquiry, that the nursing profession needs to examine values concurrently with economics. Drawing on 2 ethnographic studies of nursing practice, the authors illustrate nurses' invisible work, their invisible triaging of clients, and the invisible costs to nurses and clients. They argue that invisible work, triage, and costs are embedded in a number of values, and that if nursing is to respond to the consequences of health reform, it must examine the values inherent in economic measurement and subsequent health-policy decisions; what is invisible may go "uncounted" unless economic evaluation is informed by ethical inquiry. The authors conclude by suggesting that economic and ethical inquiry be integrated in order to foster a system that is more humane as well as more effective and efficient for all those involved in health-care delivery. PMID- 11928157 TI - Countering the deterioration and medicalization of nursing. PMID- 11928158 TI - Focus on health-care settings: the Home Care Evaluation and Research Centre. PMID- 11928159 TI - Evaluation of a weekend respite program for persons with Alzheimer disease. AB - The documented under-use of respite programs in the face of unmet family caregiver needs is puzzling. The purpose of this study was to explore family caregivers' experience with a pilot respite program of weekend care for persons with Alzheimer disease (AD) or a related dementia. The goal-free evaluation approach captured the responses of 18 family caregivers to a pilot program developed by a community nursing organization. A content analysis of the caregiver interviews identified 3 categories: caregiver self-care, relief for the caregiver, and safety and comfort of the family member. The results suggest a link between the family caregiver achieving respite and the safety and comfort of the family member. The caregivers' perspective regarding the costs and benefits of respite influences the frequency with which they use the program. Research implications are discussed. PMID- 11928160 TI - Fostering partnerships between peers and professionals. AB - One of the 5 principles of primary health care is public participation. Partnership between professionals and their clients is compatible with this principle. This article describes how professionals and peers can work together in support group interventions. Examples are cited from 3 intervention studies focused on myocardial infarction survivors and their spouses, parents of children with chronic conditions, and older women with disabilities. Each of the support groups entailed co-leadership by and partnerships between peers and professionals. The article describes the characteristics, roles, and training of peers and professionals; data collection involving peers, professionals, and participants; and pertinent intervention processes. Peers and professionals collaborated as co-leaders and partners in 21 support groups in the 3 studies. Participants expressed satisfaction with the leadership of the support groups. Moreover, professional facilitators and peer facilitators each acknowledged the important role and perspective of the other. PMID- 11928161 TI - W. Daniel Barker leadership lecture: AHA president highlights leadership challenges in today's environment. PMID- 11928162 TI - 2002 GHA Chairman installation speech: John D. Henry, Sr., FACHE casts vision for improving health care leadership. PMID- 11928163 TI - Energy dependence of photostimulable phosphor. AB - In this study, phantoms were used to illustrate the dependence of photostimulable phosphor's characteristic response on beam quality. These phantoms, consisting of sheets of acrylic and aluminum, represented an extremity, an abdomen, a skull and a chest. Images were taken with 50 to 65 kVp, 60 to 110 kVp, 65 to 80 kVp and 70 to 120 kVp, respectively. In general, the amount of resulting luminescence of the photostimulable phosphor per unit of air kerma (exposure) increased with kVp. PMID- 11928164 TI - Diagnosis and treatment of stroke. AB - This article is a review and update on stroke, the third leading cause of death in the United States. After reading the information presented, readers should be able to: Outline the etiology of stroke. Differentiate between ischemic and hemorrhagic stroke. Correlate stroke symptoms with the origin of the stroke. List risk factors and identify modifiable risk factors for stroke. Discuss the various imaging techniques used to diagnose stroke. Summarize the emergency work-up for suspected stroke patients. Identify current treatment options. PMID- 11928165 TI - HIV update. AB - Few diseases have spread so rapidly around the world or caused such severe morbidity and mortality as HIV/AIDS. After reading this review and update on the pandemic, readers will: Know the current status of HIV/AIDS in the United States and worldwide. Understand basic concepts of epidemiology and infectious disease and how these areas of study relate to HIV. Know basic concepts of immunology and the human immune system. Recognize major clinical manifestations of HIV/AIDS and their radiographic features. Understand the importance of universal precautions in protecting health care workers from blood-borne pathogens such as HIV. Understand the legal and psychosocial issues that affect people who have AIDS. PMID- 11928166 TI - Weisses blut. PMID- 11928167 TI - Demographics of the profession. AB - The R.T. with the most frequently occurring demographics is certified in one category (radiography), has an associate degree, works full time as a staff radiographer in a hospital with more than 10 other radiographers and has more than 10 years of experience. As the trend data indicate, while these demographic characteristics are still the most frequently occurring, the demographic variance has increased over time. PMID- 11928168 TI - Troubleshooting artifacts. PMID- 11928169 TI - Planning for the future. AB - Revising the strategic plan was the beginning of a multiyear initiative that will determine the path of the JRCERT. The key word in the preceding statement is beginning. The strategic plan is an ever-changing document. Although some components, such as the values statements, will stand over time, other components will change as accreditation and educational arenas change. That is the paradox of strategic planning: Remaining true to the vision, values and mission statements requires knowing when to change to keep the JRCERT aligned with the responsive to its communities of interest. PMID- 11928171 TI - Patient page. Keeping kids still during exams. PMID- 11928170 TI - Are educators ready for the future? PMID- 11928172 TI - Manuscript revisions: the team approach. AB - If authors expect and plan for revisions, editors provide clear guidelines to reviewers, and reviewers give specific, constructive suggestions to authors, quality revisions are likely to be completed. PMID- 11928173 TI - Writing a book proposal: using a question approach. AB - Most of the work of writing a book comes in the preparation of the book proposal. The process helps you carefully think through answers to several tough questions. Resources like Larsen's (1997) book, How to Write a Book Proposal, can also be a helpful guideline as you write your proposal. By the time you finish the proposal, you will be very excited about the content and prospects for the book. This is good, because when you get that acceptance, you will have to maintain your enthusiasm for the year or so you spend writing the manuscript. During the year you are writing, you will probably let some activities of daily living (ADLs) slide. After you submit the manuscript, it could take months to get your life back in order. You will start to wonder how you ever found time to write a book. You may even swear you will never write another one. Then one day, the muses will give you an idea for another book proposal. PMID- 11928174 TI - A model of women's sexual arousal. AB - A model of female sexual arousal shows the composite emotion of subjective sexual arousal, which results from conscious appraisal of sexual stimuli and their context in the presence of positive affective and cognitive feedback. Genital feedback augments the subjective arousal to a variable degree. Genital congestion can be triggered by sexual stimuli in the absence of subjective arousal. Then the congestion either is ignored or not interpreted as sexual. An anhedonic or even a dysphoric response to the sensations of genital congestion are further possibilities. This model allows for various subtypes of arousal disorder and thus facilitates a choice of therapeutic intervention. PMID- 11928175 TI - Comment on "The female sexual response: a different model". PMID- 11928176 TI - Sexual dysfunction is common and overlooked in female patients with hypertension. AB - Our objective was to investigate sexual activity, behavior, dysfunction, and satisfaction in hypertensive women. Sixty-seven patients with a mean age of 60.4 years completed a detailed questionnaire. Of these women, 81.3% had a sex partner; 42.6% had untreated sexual dysfunction with a duration of more than 5 years in 70.9% and a duration of more than 10 years in 41.7%; 5.3% initiated sexual activity; 36.6% reported less sexual activity than desired; and 54.8% reported sexual activity as important. Our study revealed highly prevalent untreated sexual dysfunction of long duration. It also showed low frequency of sexual activity in spite of the high availability of partners. There was low frequency of initiation of sexual activity. In spite of the high prevalence of sexual dysfunction, more than a third of patients reported sexual activity to be less than desired, and more than half of patients reported sexual activity as important. PMID- 11928177 TI - Repetitive male genital self-mutilation: a case report and discussion of possible risk factors. AB - Male genital self-mutilation is an infrequently reported occurrence in the medical literature. The reports describing such cases have focused mostly on surgical repair. The case reports written from a psychiatric point of view have outlined possible risk factors contributing to this behavior. Typically, reports depict cases of a single episode of self-mutilation. To date, we have found twelve cases that have addressed the specific issue of repeated episodes of genital self-mutilation. In this article, we report the case of a 49-year-old man who succeeded in castrating himself after sequential episodes of genital mutilation. We will review previous cases of repetitive male genital self mutilation and draw comparisons between those and our patient's case to arrive at common risk factors that may alert the clinician to this type of behavior. PMID- 11928178 TI - Reexploring the concept of sexual desire. AB - This article explores how sexual desire operates in ordinary life in both sexes. Fifteen considerations divided into six categories are provided: basic aspects; easily overlooked aspects; energy concepts; politically sensitive aspects; maintaining desire; and toward a definition. Sexual desire is the sum of the forces that incline us toward and away from sexual behavior. Clinicians need to conceptualize desire richly to understand its disorders. Scientists need to simplify the concept in order to conduct pharmacological research. Ideally, the limitations of both approaches must be accepted; in anyone's hands, sexual desire can be a slippery concept. PMID- 11928180 TI - Intracavernous injection during diagnostic screening for erectile dysfunction; five-year experience with over 600 patients. AB - Psychophysiological diagnostic screening (PDS) was carried out on just over 1000 consecutive patients with erectile dysfunction (ED) during 1995 through 1999. Roughly one-third did not require an intracavernous injection (ICI) because of optimal penile response during visual and tactile penile sexual stimulation. The present article deals with those other patients (n = 609) who required an ICI and completed a 24-hour-postinvestigation questionnaire (PIQ). The two ICI preparations used, prostaglandin and papaverine + fentolamine, were virtually equally effective in the doses applied, with very low and similar percentages of unwanted side effects, for example, prolonged erection. Somatic ED patients displayed the lowest penile responses to ICI, whereas psychogenic ED patients had the highest penile responses. ED patients who also suffered from premature ejaculation (PE) ejaculated significantly more readily during PDS that those without PE. An ICI following an ejaculation/orgasm could lead to an erection presumably sufficient for intromission, which indicates ICI as a therapeutic option for rapid ejaculators. Our elaborate and lengthy PDS procedure, particularly the use of penile vibration as an addition to the visual sexual stimulation, obviates the diagnostic use of the PIQ. PMID- 11928179 TI - Acute dehydroepiandrosterone effects on sexual arousal in premenopausal women. AB - The present investigation was designed to provide the first empirical examination of the effects of acute dehydroepiandrosterone (DHEA) administration on subjective and physiological sexual arousal in women. The primary purpose was to assess whether DHEA influences vaginal blood flow response in sexually functional women. Subjective (self-report) and physiological (vaginal photoplethysmograph) sexual responses to erotic stimuli were measured following DHEA (300 mg) and placebo administration in 12 sexually functional, premenopausal women, using a single-blind protocol. Acute DHEA significantly increased blood levels of dehydroepiandrosterone sulfate (DHEA-S) 30 min following drug administration but had no significant effect on either vaginal pulse amplitude responses or subjective responses to the erotic films. Acute DHEA does not appear to substantially influence sexual responding among sexually functional, premenopausal women. PMID- 11928181 TI - Helpful and harmful expectations of premarital interventions. AB - Despite the demonstrated effectiveness of premarital programs, estimates indicate that only 30% of couples use these services. This study examined the helpful and harmful aspects of premarital programs that may encourage or discourage participation. As expected, participants identified improved communication and problem solving skills as most beneficial. Disclosing secrets or past relationship issues that threaten the stability of the relationship was viewed as most harmful. Implications for the recruitment of couples and for the design and implementation of premarital programs are discussed. PMID- 11928182 TI - Brain (PET) responses to vaginal-cervical self-stimulation in women with complete spinal cord injury: preliminary findings. AB - Our recent research provides evidence that women with complete spinal cord injury (SCI) at the midthoracic level show perceptual responses to vaginal and/or cervical self-stimulation (for example, pain suppression and sexual response, including orgasm). On the basis of studies in laboratory rats, we hypothesized that the vagus nerves provide a sensory pathway from the vagina, cervix, and uterus directly to the brain in women. To test this hypothesis, we performed a PET-MRI study on two women with complete SCI and 1 woman with no injuries. Whereas control foot stimulation of the women with SCI did not activate the somatosensory thalamus, cervical self-stimulation increased activity in the region of the nucleus of the solitary tract, which is the brainstem nucleus to which the vagus nerves project. These preliminary findings suggest that the vagus nerves can convey genital sensory input directly to the brain in women, completely bypassing SCI at any level. PMID- 11928183 TI - [New possibilities in the treatment of Meniere's disease]. AB - The results of the treatment of Meniere's disease by intratympanic instillation of gentamicin in 32 patients over a 24-months period were analyzed. The receded vestibular function was found in 96.8% patients, whereas in the remaining 3.2% pronounced vestibular hypofunction was observed. One month after the treatment vertiginous attacks disappeared in all the patients. However, three months after the treatment infrequent episodes of mild degree vertigo and mild unsteadiness that did not require additional intratympanic gentamicin application appeared in 15% of the patients with Meniere's disease. The vertiginous attacks ceased or were pronouncedly reduced in 85% of the patients. The unstable vestibular hypofunction was successfully converted into stabilized, centrally compensated areflexia. Intratympanic gentamicin exerted better results than the other invasive therapies for intractable Meniere's disease. PMID- 11928184 TI - [Effect of uremia and peritoneal dialysis on peritoneal mesothelial cells]. AB - The aim of the study was to investigate the morphology of mesothelial cells of the peritoneum of patients with terminal renal failure (TRF), taken by the biopsy immediately before the onset of peritonal dialysis (PD), and to compare it with the findings in patients with PD. The samples were prepared in the way standard for light microscopy and transmission electron microscopy. In patients with TRF intracytoplasmatic inclusions could be observed, unusual protrusions of mesothelial apical surfaces, deformation of mesothelial cells and their detachment from the basal membrane, as well as the dilatated cisternae of granulated endoplasmatic reticulum with filamentous structures in some of them. In patients on PD cytoplasmic protrusions of different shapes and contents were observed at the surface of mesothelial cells, multiplication of basal membrane, occurrence of young forms of mesothelial cells as well as the detachment of those cells from the basal lamina. PMID- 11928185 TI - [Tumor markers in pleural effusions in bronchogenic carcinoma and tuberculosis]. AB - Concentrations of carcinoembryonic antigen (CEA) and carborhydrate antigen (CA) 50 were measured in pleural effusion and sera of 57 patients with bronchogenic carcinoma and in 73 patients in whom the effusion was the sequela of tuberculous pleurisy. In the group with bronchogenic carcinomas, planocellular was confirmed in 19, microcellular in 17, macrocellular in 2, and adenocarcinoma in 18, while in 1 patient it was not possible to determine the histopathologic structure. The diagnosis of pleural disease was established upon the cytologic examination of the effusion and histopathologic examination of the pleural sample obtained by blind percutaneous needle biopsy or following pleuroscopy. CEA concentration in the sera of patients with bronchogenic carcinoma was significantly higher than in the patients with tuberculosis (p < 0.001), with sensitivity of 44% and ideal specificity and positive predictive value of 100%. In the same group highly significant difference of mean values of CEA concentrations in pleural effusion (p < 0.001), was also found with sensitivity of 60%, significant specificity of 99% and positive predictive value of 97%. CA 50 concentrations in the sera of patients with lung carcinoma were significantly higher than those in the sera of patients with tuberculous pleurisy (p < 0.05), and the sensitivity was 50%, while the specificity was 94% and positive predictive value was 75%. Significantly higher was also the value in the pleural effusion (p < 0.05), but the sensitivity was slightly lower--40%, but specificity was favorable as well as the positive predictive value (94 and 86%, respectively). The results indicate the significance of the determination of CEA and CA 50 in the sera and pleural effusion in the differentiation of malignant from tuberculous pleural effusion. PMID- 11928186 TI - [Correlation of size of the primary tumor and axillary node status with the p53 tumor suppressor gene in carcinoma of the breast]. AB - Correlation of standard pathomorphological prognostic parameters, primary tumor size and axillary nodal status with new prognostic factor in breast carcinoma: tumor suppressor gene p53 was analyzed. The studied sample included 65 women who underwent surgery for breast carcinoma at the Surgical Clinic of Clinical Center Banja Luka, from January 1st 1997 till January 1st 1999. Statistical data analysis was performed and correlation of prognostic factors was determined. The majority of authors in this field agree that the primary tumor size and axillary nodal status are the two most important prognostic factors. These factors are the best predictors of prognosis and survival of women who had the tumor and were operated on. Tumor markers were immunohistochemically determined in the last ten years and, according to the majority of authors, are still considered the additional or relative prognostic factors in breast carcinoma. Their prognostic value and significance increase almost daily. Most frequently determined tumor markers are bcl-2, pS2, Ki-67 and p53. There was a positive, directly proportional relationship between primary tumor size and tumor suppressor gene p53, but there was no positive correlation between the axillary nodal status and tumor suppressor gene p53. Significance of determination of new tumor markers as the prognostic factors was emphasized. These markers represent a powerful tool in the early detection and prevention of breast carcinoma. PMID- 11928187 TI - [Treatment of aneurysms in the anterior cerebral circulation]. AB - OBJECTIVES: To analyze the outcome of either surgical or conservative treatment of patients with aneurysms on cerebral arteries. DESIGN: Retrospective study on 114 patients (89 operated and 25 not operated). METHODS: Clinical state was graded from 0 to V, according to Hunt & Hess (HHG), and the treatment outcome was defined as favorable or poor, according to the modified Glasgow Outcome Score. The outcome was correlated with the type of treatment (operative or conservative), clinical state and aneurysmal localization. RESULTS: Aneurysm was localized mostly on the anterior communicating (33.6%) and middle cerebral arteries (32.8%) and the patients were mostly in HHG II or III (34.4% and 25.2%). HHG after the aneurysmal rupture did not depend on the aneurysmal location (p > 0.05). Favorable treatment outcome was noted: in 74.1% of all operated and in 60% of all conservatively treated patients (p > 0.05); in 81.6% of operated and in 33.3% of not operated patients with HHG = II-III (p < 0.01); in 78.8% of aneurysms of the middle cerebral artery and in 66.7% of those of the anterior communicating artery (p > 0.05); in 73.1% of patients with HHG = III and in 25% of patients with HHG = IV (p < 0.01). CONCLUSIONS: Clinical state after the aneurysmal rupture did not depend on its localization. Results were better after the surgical, than after the conservative treatment. Outcome after the surgery depended on the clinical state of the patient, but not on the aneurysmal localization. PMID- 11928188 TI - [Use of disodium pamidronate in patients with bone metastases in patients with pulmonary carcinoma]. AB - Disodium pamidronate (Aredia), a drug of the bisphosphonate group, was administered to 20 patients with pains in the bones due to secondary deposits of lung cancer. The aim of the study was to investigate the analgesic effect of pamidronate to osteolytic metastases of lung cancer in the bone. In 16 (80%) patients non-small-cell lung cancer was diagnosed, and in 4 (20%) patients small cell-lung cancer was confirmed. Intensive disseminated pains in the bones were present in all the patients. Metastases in the skeleton were confirmed by radiography and scintigraphy of the bones. Initial values of calcium in the plasma were determined in those patients. In 11 (55%) patients, initial values of calcium in the serum were normal, and in 9 (45%) patients, they were elevated. Patients with normal calcium values received 30 mg of pamidronate in 250 ml of normal saline, and patients with hypercalcemia received 45-60 mg in 500 ml of normal saline. Analgesic effect of pamidronate was present in 12 (60%) patients, and the completely painless state was achieved in 4 out of 12 patients. Evaluation of the pain was done by questionnaire, using a simple, descriptive scale. In all 9 (100%) patients with hypercalcemia, values of calcium in plasma were normalized. Pamidronate exerted favorable analgesic effect in the case of metastases of lung cancer in the bones in more than 50% patients. Satisfactory results were also achieved in the patients non-responsive to palliative therapy by irradiation. This enables the use of opiates in lower doses. PMID- 11928190 TI - [Initial masseter muscle fatigue in maximal voluntary teeth clenching]. AB - Fatigue of striated muscles is defined as the impossibility to generate the expected or required force during the repeated contraction. During the maximal voluntary teeth contraction in the position of central occlusion the initial fatigue in masticatory muscles during the isometric contraction occurs. If a person can clench its teeth continuously and voluntarily it has a diagnostic significance since the peripheral fatigue is that important factor in the masticatory muscles activity, which is in direct correlation with the function of the masticatory system. The aim was to compare the obtained results of the initial fatigue of masseter muscles during the maximal voluntary teeth contraction in subjects with naturally healthy intact dentition and subjects with a pair of new full dentures. The investigation comprised 20 subjects with healthy stomatognathic system of the skeletal class I by Angle. Comparison of the values of the obtained results was performed electromyographically by synchronous registration of action potentials of masseter muscles. Results of the investigation of the onset of initial fatigue of masseter muscles in the subjects with natural healthy intact dentions showed lower values, i.e., faster development of the muscular fatigue (31.5 s) compared to the subjects with a pair of new full dentures (44.5 s). PMID- 11928189 TI - [Intradermal tests with vasomotor agents in chronic noninfectious rhinitis]. AB - The aim of this prospective study was to examine the skin reactivity to four vasomotor agents in chronic, non-infectious rhinitis patients, and to determine whether non-allergic rhinitis (NAR) patients differ from allergic rhinitis (AR) patients. Seventy four patients with NAR and 44 with AR were subjected to intradermal testing with papaverine (5 mg/ml), metacholine (0.02, 0.2 and 2.0 mg/ml), histamine (0.01, 0.1, 1.0 and 10.0 micrograms/ml), compound 48/80 (0.01, 0.1, 1.0 and 10.0 micrograms/ml) and saline. It was found that the frequency of pathological skin reactivity to papaverine in the patients with NAR (25/74) was significantly greater (p = 5.0 x 10(-3)) then in the patients with AR (4/44). No significant inter-group difference in skin reactivity to metacholine, histamine, compound 48/80 and saline was observed. The frequency of the total pathological skin reactivity to vasomotor agents, singly and in combinations, in patients with NAR (80%) was significantly greater (p = 0.03) than in patients with AR (61%). These findings suggested that the pathological skin reactivity to papaverine, metacholine, histamine and compound 48/80 was a feature of chronic, non infectious rhinitis patients and it was more frequently associated with non allergic than with the allergic etiology of rhinitis. PMID- 11928192 TI - [Cytomegaloviruses and atherosclerosis]. PMID- 11928191 TI - [Pain and excitatory amino acids]. PMID- 11928193 TI - [Dolichoarteriopathy--kinking and coiling of the carotid arteries]. PMID- 11928194 TI - [Role of endovascular procedures in the treatment of cerebral ischemia]. PMID- 11928195 TI - [Recurrent episodes of focal cerebral ischemia in a female patient with mitral valve prolapse and migraine headache]. AB - Migraine is episodic, paroxysmal disorder where the headache represents the central symptom and is followed with different combinations of neurologic, gastrointestinal and vegetative changes. Not until the diagnostic procedures were developed, ischemic lesions were verified even in the patients with ordinary migraine. This is a report of a patient with migraine headache, followed twice by verified episodes of temporary ischemic attacks and verified focal ischemic lesion of cerebral parenchyma. The mitral valve prolapse was also detected. This all imposed the administration of combined prophylactic antimigrainous and anticoagulant therapy as an imperative because of the risk of the development of repeated ischemia of cerebral tissue. This association also confirmed an opinion that migraine is a wider disorder with the dominant dysfunction of limbic system. PMID- 11928197 TI - Working with dreams in a bereavement therapy group. AB - There is ample theory and research about group therapy, dream work, and bereavement as separate subjects. However, there is little written specifically about utilizing dream work in bereavement therapy groups. Using the Foulksian group analytic model, dreams in one particular bereavement group (for parents of children killed in a terrorist action) were interpreted in such a way as to help members access deep unconscious feelings. This helped facilitate a fuller and more complete mourning process. The analytic, dream interpretive activity also helped overcome resistance in the group-as-a-whole and thereby facilitated movement through group development phases. PMID- 11928196 TI - [Laza K. Lazarevic (1851-1891)--part 2]. PMID- 11928198 TI - Women in group and women's groups. AB - Female development is characterized by the introjection of maternal and paternal objects, which creates a rich internal world for women. However, the feminine tendency to become diffused in identifications sometimes results in women feeling overpowered in mixed-gender groups. The author advocates women's groups for the traditionally "feminine" woman, that is, the woman who has a rather impoverished sense of self in terms of her internalized objects. Women's groups are also helpful during adolescence, pregnancy, and menopause, milestones of female development. These are times when women are in profound crisis and may find it difficult to speak about these quintessentially feminine events in the presence of men. PMID- 11928199 TI - A dualistic model for group treatment of alcohol problems: abstinence-based treatment for alcoholics, moderation training for problem drinkers. AB - This article provides a clinical framework for a dualistic group treatment model: abstinence-based treatment for alcohol dependent individuals and moderation training for problem drinkers. The major premises of these models are set forth, compared, and contrasted. An integrated approach for synthesizing these models is then detailed along with suggestions for clinical implementation and management of countertransference. PMID- 11928200 TI - Existential issues in group psychotherapy. AB - Existential issues in group psychotherapy derive from existential thought both as a philosophy and as a value system. Its origins derive from the weakening of traditional values and the growing alienation of man from himself. The unique features of existentialism can be applied to all forms of therapy. These features are universal to humankind. They are finiteness, aloneness, guilt, responsibility, and freedom. In including existential concerns as part of group psychotherapy, therapist and patients move more closely to bilateral relationships and subjective interactions. PMID- 11928201 TI - The demonstration group: a tool for observing group process and leadership style. AB - Demonstration groups, a major modality for observing group process, have not been addressed in the group psychotherapy literature. This article defines the demonstration group and describes and discusses its various components: the volunteers (group members) and their recruitment, the demonstration group itself, group member debriefing, and the didactic component. Discussion of the physical setting, group agreements, and boundary considerations, as well as leadership tasks, challenges, and pitfalls are illustrated with examples. Leadership tasks in demonstration groups are compared with those in therapy groups. Guidelines that are likely to promote successful demonstration group experiences are provided. The leader's facility with the unconscious defense mechanism of projective identification is stressed. PMID- 11928202 TI - Videotape resources for group psychotherapists: a 5-year retrospective. AB - This review provides a discussion of the 14 videotape reviews that have appeared in the International Journal of Group Psychotherapy from 1996 through 2000. This retrospective analysis describes the recommendations made by the review authors concerning ways in which future video products may be enhanced to improve their instructional value. These recommendations include topics that might be given greater emphasis in future efforts as well as elements within the format of such tapes that aid learning and discussion. PMID- 11928203 TI - Developmental processes in women's groups in Italy. AB - The observation of women's groups in Italy has made it possible to outline specific developmental processes. Clinical illustration also suggests the emergence of specific group areas such as "interchangeability," which are not seen in mixed-gender groups. The safe and confident atmosphere of interchangeability stimulates the relaxation of defences and self boundaries. As a consequence, it allows profound exchanges, which have an immediate impact in enhancing the sense of value and self-esteem of group members. Further therapeutic potentialities of "interchangeability" are described. Women in the here and now acquire the strength necessary for reexamining and psychoanalytically working through unconscious unresolved gender identifications and consequent problems in present relationships. PMID- 11928204 TI - Emotional incest in group psychotherapy. PMID- 11928205 TI - Response to "The therapist's anxiety and resistance to group psychotherapy". PMID- 11928207 TI - Is worker age a simple demographic variable? AB - Birth-date-based chronological age is often used as a measure of worker ability. In the near future, employers will be required to improve their ability to assess the impact of aging on workers. The literature suggests a number of ways one might do this. Personal perceived age and work ability assessment are two promising alternative ways to assess worker age changes. In support of these alternatives and as a preface to the other papers presented in this journal, this paper suggests that personal perceived age reports differ from chronological age, are reliable, and appear to be worthy of further consideration. PMID- 11928208 TI - Age differences in learning maintenance skills: a field study. AB - The effects of age and previous relevant experience on learning anxiety, strategies, and performance were studied in 43 workers aged 25 to 49 during a 1 week maintenance vocational training course. The results showed that increased age was associated with higher training-related anxiety as measured at the beginning of the course. However, no age difference could be found in the level of knowledge assessed after 3 days of training. This was confirmed by another problem-solving-type test that took place on the last day. Previous experience had no effect on anxiety, and it did enable us to predict higher scores for the first test but not for the second one. Analysis of behavior strategies revealed that older trainees consulted and annotated the course material more often than the younger ones during the learning process. The results are discussed in relation to those obtained in previous laboratory and field studies on the same subject. PMID- 11928209 TI - Chronological and subjective age in relation to work demands: survey of Croatian workers. AB - The aim of the study was to investigate chronological and subjective age and gender differences in terms of the physical, mental, and social demands of work. Subjects, 88 women and 25 men, were workers in a medical setting who completed the Survey of Health Care Professionals. Subjects' reports of effort required by their work, tiredness resulting from their work, and personal skills needed to complete their work were used to assess the physical, mental, and social demands of work. Subjective age measures included cognitive age, desired age, and comparative age. Results showed that, compared to men, women reported being more tense as a result of the work. Age differences were found in the physical and mental skills required by the work. Older workers had lower estimates of their own skills than younger workers. Three measures of subjective age showed different patterns of relationships with aspects of work demands. PMID- 11928210 TI - Personal age and assessment of work stress in Polish nurses. AB - Workers' perception of age and work stress was examined within the context of aging and work. Nurses from selected hospitals in a large Polish town completed the Survey of Health Care Professionals to assess their personal age and on-the job and off-the-job stressors. The assessment of work performance, effort, and abilities associated with work demands and tiredness after the workday was compared between groups of subjects differing in the magnitude and direction of discrepancies between felt age and chronological age. The results showed that, compared to others, nurses feeling older than their chronological age assessed themselves as putting forth the highest effort to meet their work demands, reported more tiredness after the workday, and reported the lowest work abilities. PMID- 11928212 TI - Physical requirements associated with the work of aging workers in the European Union. AB - Physical requirements of work should decline with advancing age because the dimensions of physical work capacity of individuals decline with age, too. The Second European Survey on Working Conditions in 1995/1996 was used to study the prevalences (%) of poor work postures, handling heavy loads, and repetitive work in the representative sample of employed people in the 15 European Union member states. Exposure to physical requirements was still common in the European Union. Nearly 50% of older workers (age 45 years or more) were exposed to repetitive work, about 30% had poor work postures and 15% to 20% were handling heavy loads at least half of the working time. The physical requirements were as common for older and younger (less than 45 years of age) workers. The physical requirements were most common in Mediterranean countries and least common in the northern part of Europe. The differences in prevalence rates of exposures between the countries were large, from three- to fivefold. Favorable differences between older and younger workers were more notable for older men than for older women in poor work postures and repetitive work. However, older women had less handling of heavy loads than older men. The results showed that the need for better adjustments of physical requirements with age is still very relevant in the European Union and demands urgently appropriate measures in working life. PMID- 11928211 TI - Work ability, age and its perception, and other related concerns of Ukraine health care workers. AB - A sample of 250 health care workers aged 18 to 68 (mean = 32.5 years) completed the Survey of Health Care Professionals. Self-ratings of their social skills, mental capacity, and physical capability corresponded to their ratings of work demands. Physical tiredness and tension were rated higher than mental tiredness. Worker age did not affect self-ratings of work performance, but physical and mental tiredness increased with increases in the age that one felt. The younger participants felt compared to their calendar ages, the better the level of current work ability they reported. The main concerns of workers were connected with off-the-job factors, most likely caused by the economic crisis and unfavorable ecological conditions in Ukraine. More than half of the participants were quite a bit or extremely concerned with changes in the cost of living, water quality, food safety, and radiation. The variable most closely related to these concerns is the discrepancy between calendar age and how old one feels. Coping strategies of workers can be related to sleeping, entertainment, and other off the-job activities. These behaviors are related to the discrepancy between calendar age and how old one looks and feels, as well as felt age. PMID- 11928213 TI - Unveiling factors that contribute to functional aging among health care shiftworkers in Sao Paulo, Brazil. AB - The aims of this study were to evaluate aging factors associated with work stressors, work ability, and the quality of living conditions, among health care personnel. A cross-sectional study was conducted among 176 health care shiftworkers. Two health survey questionnaires (Tuomi et al., 1997, Scandinavian Journal of Work, Environment and Health, 17(Suppl 1), 67-74; and Tepas, 1996, unpublished instrument) were completed and ergonomic work analyses (Rohmert & Landau, 1983, A new technique for job analysis, London and New York: Taylor & Francis) were carried out at the emergency wards. Main concerns about exposure at the workplace were changes in equipment and technology, transportation, and changes in employer policies. Main concerns about off-the-job conditions were personal safety, increases in the cost of living, food safety, and water and air quality. 81.7% scored adequate (> 36.5 points) in the Work Ability Index, and considered themselves having adequate current work ability to cope with physical, mental, and social demands. The most frequently reported diseases were musculoskeletal disorders and minor emotional problems. PMID- 11928214 TI - How do concepts of age relate to work and off-the-job stresses and strains? A field study of health care workers in five nations. AB - Health care workers from five nations (Brazil, Croatia, Poland, Ukraine, U.S.A.) completed the Survey of Health Care Professionals, which includes measures of on the-job and off-the-job conditions/concerns, as well as ratings of work-related strains, self-ratings of work performance and work ability, and measures of chronological and psychological age. Consistent with previous research, workers generally reported that they felt, looked, acted, and preferred to be younger than their chronological age. In addition, feeling old relative to one's chronological age was positively associated with several indicators of job related strain for several nations, replicating findings reported earlier in a U.S. manufacturing sample. Significant differences were observed among nations on all of the psychological age variables measured in this study. Differences among nations were also observed for levels of concern with work conditions and off-the job conditions. Furthermore, examination of relationships of age variables with work/off-the-job stressors and work strains/outcomes revealed different patterns of relationships among the five nations participating in the study. Findings point to the need for additional attention to psychological age variables in studies of work and aging, with particular emphasis on cultural and socioeconomic conditions that may influence their interplay. PMID- 11928216 TI - Safety and risk for RNs on home visits. PMID- 11928215 TI - Prioritizing ergonomic research in aging for the 21st century American workforce. AB - The Bureau of Labor Statistics predicts that by 2008 Americans aged 55 and over will represent the greatest annual growth rate and will occupy 30% of the American population. Additionally, by 2008 civilian labor force participation rates for the 55 and older group will grow to 36.8%, a 6.5% increase over the participation rates for this group for 1996, with the 55 to 64 age group expected to add 7.3 million workers. The predicted median age of the labor force for 2008 is estimated at 40.7 years, an age not approached because the median age peaked at 40.5 years in 1962. Coupled with this aging profile, employment in professional specialty occupations will increase the fastest and add the most jobs in the decade leading to 2008. Within the professional specialty professions, the majority of the employment increases are expected to occur in the service industry division. The purpose of this paper is to merge demographic data with ergonomic and human factors data and predictions to explore areas of ergonomic research that will aid in keeping the aging workforce, and those with whom they interact, healthy in their jobs and keep the companies for whom they are employed competitive. Although some ergonomics research has been conducted in all the listed areas of expected growth, the paper reexamines the job demands in each sector, with a focus on the needs of the aging workforce. PMID- 11928217 TI - A primer on rural nursing. PMID- 11928218 TI - New guidelines for AARN elections. PMID- 11928219 TI - [Potential role of the early response gene c-Myc in the loss of differentiation and function of pancreatic beta cells submitted to chronic hyperglycemia]. AB - Chronic hyperglycemia exerts deleterious effects on glucose-induced insulin secretion by pancreatic beta cells. In 90% pancreatectomized rats, chronic hyperglycemia induces beta cell hypertrophy and loss of differentiation associated with increased expression of the early response gene c-Myc. Hyperglycemia also stimulates c-Myc expression in vivo in islets from glucose infused rats and in vitro in cultured rat islets. This effect requires the elevation of cytosolic Ca2+ concentration produced by glucose. Our results suggest that overstimulation of c-Myc expression by chronic hyperglycemia may be the cause of beta cell hypertrophy and loss of differentiation and function observed in animal models of type 2 diabetes. PMID- 11928220 TI - [New research on the significance of polymers in pharmaceutical formulations]. AB - During these last few decades, a lot of work has been made in pharmaceutical area in order to control the drug delivery from various pharmaceutical dosage forms. The use of polymers in pharmaceutical technology have led to the development of the first drug delivery systems proposed in order to prolong or to delay the drug delivery, or to enhance drug release for drugs showing bioavailability shortcomings. The wide range of polymers available for pharmaceutical use, their low reactivity towards drugs and other formulation ingredients and their safe nature, have permitted a widespread use of polymers to improve manufacturing processes or for the formulation of pharmaceutical dosage forms for various administration routes. More over, the preparation of new polymeric materials by the synthesis of new polymers with unique properties or by the modification of available natural or synthetic polymers, offer to the formulator a wide range of applications in order to optimise the drug delivery for each specific case. PMID- 11928221 TI - [Enantiomeric separation of drugs with capillary electrophoresis with cyclodextrins]. AB - General strategies for the development of capillary electrophoretic methods for the enantiomeric separation of basic, acidic or neutral drugs have been developed. For all kinds of compounds the use of a buffer made of 100 mM phosphoric acid adjusted to pH 3 or 5 with triethanolamine, containing cyclodextrin (CD) derivatives as chiral selectors, is recommended. Five b cyclodextrin derivatives were found to be particularly useful for enantioseparations: dimethyl-beta-CD (DMCD), trimethyl-beta-CD (TMCD), hydroxypropyl-beta-CD (HPCD), carboxymethyl-beta-CD (CMCD) and sulfobutyl-beta-CD (SBCD). Two different strategies are proposed, depending on the acidic or basic character of the analytes. Using these strategies, 96% of the 50 drugs examined as model compounds could be fully enantioseparated, which confirmed the usefulness of capillary electrophoresis in order to obtain complete enantiomeric separation as quickly as possible and with a minimum consumption of chiral selector. PMID- 11928222 TI - [Involvement of polynuclear eosinophils and of type TH2 immune responses in allograft rejections]. AB - In the present work, we showed that mouse alloreactive Th2 type lymphocytes are able to acutely or chronically reject the MHC class II disparate skin allografts. Acute skin allograft rejection is characterized by IL5 production and eosinophils infiltration. IL5 neutralization in the absence of CD4 T cell cytotoxicity prevented the eosinophil infiltration and the acute rejection in the large majority of recipients. Chronic rejection was characterized by graft fibrosis and vasculopathy. Whereas IL4 and IL5 neutralization prevented eosinophil infiltration and graft fibrosis, the sole IL4 neutralization completely prevented vasculopathy, which was eosinophil and IL5 independent. Together, these observations illustrate the mechanisms used by Th2 alloreactive cells in the allograft rejection process. PMID- 11928223 TI - [Kappa-B nuclear factor and apoptosis of cancerous cells]. AB - The transcriptional factor NF-kappa B is a key regulator of many biological processes such as the immune response, inflammation, development, cell proliferation and apoptosis. We investigated the role of NF-kappa B in the regulation of the pro-apoptotic protein Bax. We observed increased Bax proteins expression in several cancer cell lines stably expressing a mutated I kappa B alpha that blocks NF-kappa B activity. Transient transfection experiments showed that NF-kappa B inhibited p53-dependent transactivation of the bax promoter, but this effect was not released by concomitant expression of several cofactors including CBP/p300. We also identified, for the first time, an NF-kappa B binding site in the bax gene promoter but the mutation of this site did not affect NF kappa B induced inhibition of transcription. The mechanisms responsible for NF kappa B inhibition of the bax promoter thus remain to be elucidated. PMID- 11928224 TI - [Development of new antineoplastic agents: 10 years of active clinical research marked by the evolution of treatment, from an empirical to a rational approach]. AB - In addition to well-known cytotoxic agents in the treatment of cancer, promising novel cytotoxic and biologic agents are being investigated. The combination of classical cytotoxic drugs or radiotherapy, along with these new molecules specifically targeted at numerous points in intracellular signalling, cell cycle control, and the cancer cell environment, represents our best hope for improved outcome in systemic cancer therapy. This report summarizes the phase I clinical studies with these novel anticancer agents, performed and supervised the last ten years by the author at the chemotherapy unit of the Jules Bordet Institute. PMID- 11928225 TI - [Experience of 6 years with multiple sclerosis]. AB - The MS clinics of Brussels and Charleroi have been in existence six years. This paper presents a review of our experience, during this period, of the current MS treatments, as well as a follow-up of those patients who participated in various European clinical studies. PMID- 11928226 TI - [Lung transplantation and mitochondrial function]. AB - The mechanisms of cellular lesions induced by lung ischemia and reperfusion are not fully understood and, in particular, the consequences of pulmonary ischemia and reperfusion injury on mitochondrial function have not been previously investigated. Therefore, we studied the respiratory function of isolated pulmonary mitochondria in a swine model of lung ischemia and reperfusion. We demonstrated that prolonged hypothermic (4 degrees C) ischemia induces significant lesions of the mitochondrial respiratory chain, particularly if ischemia is followed by normothermic reperfusion. These results should be integrated in the cellular alterations induced by the ischemia-reperfusion injury. In another swine model mimicking controlled non-heart beating donors, we demonstrated that thirty minutes of cardiac arrest do not promote significant alteration of the mitochondrial respiratory function. In contrast, forty-five minutes of cardiac arrest, induced significant mitochondrial lesions. This pulmonary tolerance to normothermic cardiac arrest might be explained by the presence of air in the lung airways, allowing some aerobic metabolism after circulatory arrest. These results suggested that lung grafts might be harvested from non-heart beating donors after thirty minutes of cardiac arrest, significantly increasing the pulmonary graft pool. PMID- 11928227 TI - [A specialty that deals with breast cancer. Breast pathology hospital units]. AB - Breast cancer has become a veritable "plague" on Western European civilization to the point where one in ten women will likely develop a mammary gland neoplasia in the future. From a medical and health policy perspective, we must be prepared to deal with this problem. Undergraduate and post-graduate education of professionals and the creation of specific units are some of the solutions for fighting this epidemic. PMID- 11928229 TI - [Application of the principle of precaution to medicine]. AB - The principle of precaution prescribes taking immediate action to avoid serious damage, rather than waiting for proof of the danger. It was extended to medical field, without taking account of the specificity of medicine, as well in individual medicine as in public health. Radical application of this concept could lead to serious repercussions. After analysis, it will considered as an expression of the historical confrontation between the science and the public opinion. PMID- 11928228 TI - [The right to be aborted]. AB - This lecture deals with the difficult problem of so called "wrongful life". Two Court decisions have been issued recently in France and lead to a very ambiguous situation since they differ dramatically. In the "Perruche affair" where a diagnosis of rubella was not done, the parents prevented to get an interruption of pregnancy, and the child severely affected, the Court (Cour de Cassation, i.e. Supreme Court in charge of civil affairs) decided to compensate the child for "not having be aborted". In the "Quarez affair" where a diagnosis of Down syndrome was not done, the parents prevented to get an interruption of pregnancy, the Court (Conseil d'Etat, i.e. Supreme Court in charge of "administrative" affairs, affairs opposing individuals to state administration since the hospital in cause was belonging to the public sector) decided not to compensate directly the affected child. The consequences of these opposite decisions are analysed according to medical, legal, ethical and economic considerations. PMID- 11928230 TI - [Stromelysin-1 in inflammatory disease: significance, specificity, and regulation elements]. AB - Stromelysin-1 or matrix metalloproteinase-3 (MMP-3) is a key enzyme of the degradation of the extracellular matrix in rheumatoid arthritis. MMP-3 is indeed induced by pro-inflammatory cytokines and may in turn activate other metalloproteinases. MMP-3 is a synovial parameter reflecting the local inflammatory reaction induced by inflammatory cytokines. It is not specific for rheumatoid arthritis nor for the erosive capacity of an arthropathy, which does not exclude its role in articular degradation. Serum MMP-3 is increased in diseases characterized by synovitis but also by steroid therapy. Although it is not disease-specific, serum MMP-3 could be useful as a synovial-derived marker of local inflammation in rheumatoid arthritis, in parallel to CRP which is a systemic marker of inflammation. Furthermore, early determination of serum MMP-3 could constitute a new tool predictive of the disease activity and the therapeutic response in rheumatoid arthritis. PMID- 11928231 TI - [Orthognathic surgery, master-piece of maxillo-facial surgery]. AB - Orthognathic surgery is this field of the maxillofacial surgery which aims to reposition the jaws or some segments of these jaws when masticatory dysfunctions are evident. This tridimensional repositioning in the craniofacial skeleton allows to restore the masticatory function by means of osteotomies, which must be followed either by preoperative simulated bony displacements or by callus bone distraction. Not only are the functional benefits evident on the dental, articular and neuromuscular levels, but also a facial esthetic harmony can almost be obtained. PMID- 11928232 TI - Joint Commission wants to see new effort on terrorism plans. PMID- 11928234 TI - Credential your allied health professionals. PMID- 11928233 TI - Health plans report quality improvements. PMID- 11928235 TI - QI project: get babies home fast, yield savings. PMID- 11928236 TI - Office CM program draws praise from physicians. PMID- 11928237 TI - Don't let language be barrier to care. PMID- 11928238 TI - Compensation monitor. Primary care physicians' compensation better reflects productivity. PMID- 11928239 TI - He wants AdvancePCS to manage more than drugs. Interview by Patrick Mullen. PMID- 11928240 TI - Defined contribution: threat ... or fad? AB - Sensing an invasion of their territory, MCOs are jumping into a market forged by a group of upstarts. The development renews a fundamental debate about the juxtaposition of consumer involvement, cost containment, cost shifting, and quality of care. PMID- 11928241 TI - The Department of Managed Care. AB - California sets a precedent again, with a bold effort to regulate the HMO industry. While the new Department of Managed Health Care is facing down 50,000 consumer gripes monthly, director Daniel Zingale intends to keep administrative red tape from overwhelming plans, patients, providers, and his department. PMID- 11928242 TI - Some lessons to be learned from Canadian health system. PMID- 11928243 TI - DM vendors start to address costs created by comorbidities. PMID- 11928244 TI - An in-office diagnostic procedure to detect dermatophytes in a nationwide study of onychomycosis patients. AB - PURPOSE: To evaluate in-office dermatophyte test medium (DTM) culture as an alternative to traditional laboratory fungal culture for confirming a diagnosis of onychomycosis, and to determine the prevalence of dermatophytes as a cause of onychomycosis in patients not participating in a clinical trial. DESIGN: This nationwide multicenter prospective study enrolled 1100 adult patients with suspect onychomycosis. DTM and laboratory fungal culture results were compared for individual patients. METHODOLOGY: The 310 participating physicians obtained patient nail-bed specimens and divided them for testing by both diagnostic methods. The paired results of the two culture methods were compared using the kappa statistic. PRINCIPAL FINDINGS: Paired culture results were available for 975 of the 1100 enrolled patients. DTM results agreed with central laboratory cultures in 70 percent of cases. The kappa value of 0.40 indicated a moderate degree of correspondence between the two testing modalities. Overall, DTM culture indicated a dermatophyte in 616 patient specimens (56 percent) and central laboratory culture identified a dermatophyte in 528 of the specimens (48 percent). For the entire study population, dermatophytes were identified in 93 percent of the positive central laboratory cultures, confirming that dermatophytes caused the vast majority of the infections. The cost of each DTM culture was approximately $1, compared to $25 for each laboratory fungal culture. CONCLUSION: This study demonstrates that the in-office DTM culture for diagnosing onychomycosis has comparable utility to the traditional laboratory fungal culture, is less expensive, and yields faster results. PMID- 11928246 TI - 'Team approach' to contract talks means that nobody feels cheated. PMID- 11928245 TI - Getting patients in the door faster can boost satisfaction, outcomes. PMID- 11928247 TI - Managed care outlook. Employees increasingly inclined to enroll for benefits via Internet. PMID- 11928248 TI - Is your cardiovascular program meeting goals? PMID- 11928249 TI - Retention of highly productive personnel now at crisis proportions. PMID- 11928250 TI - How will hospitals choose auditors? PMID- 11928251 TI - Indiana [correction of Indian] Heart Hospital will be all digital hospital without paper records. PMID- 11928252 TI - Infant mortality rates are proxies for use of neonatal intensive care units in hospitals. PMID- 11928253 TI - Clinical journals' Web sites offer a wealth of data to hospital planners and strategists. PMID- 11928254 TI - Messenger models still are antitrust problems for providers. PMID- 11928255 TI - [Hypertension and impotence. Which potency drug is the most appropriate here?]. PMID- 11928256 TI - [Complaint of a family physician. "For my patients I am only a substitute doctor"]. PMID- 11928257 TI - [Become a nutrition counselor. Your patients will appreciate it (interview by Christine Vetter)]. PMID- 11928258 TI - [Reflux disease. Don't annoy your patients with ineffective general measures!]. PMID- 11928259 TI - [Acute, chronic or physical urticaria. What causes the hives?]. AB - Urticaria is characterized by strongly pruritic wheals and in some cases, angioedema. On the basis of a careful case history, the type of urticaria presenting can usually be determined. In the case of the acute form, no complicated diagnostic work-up is necessary. Treatment is symptomatic: the use of ASA should be avoided, while an acute infection requires treatment. When there is recurrent angioedema, chronic urticaria and physical forms, a multitude of triggering factors must be considered. In many cases, symptoms may persist for years. Symptomatic treatment with antihistaminics is often inadequate, but long term treatment with glucocorticoids should be avoided on account of their side effects. For this reason, further diagnostic work-up is usually necessary and, where indicated, treatment should be initiated by a dermatologist with specialist training in allergology. PMID- 11928260 TI - [From oil bath to Lactobacillus. How does one treat neurodermatitis?]. PMID- 11928261 TI - ["Pollen-associated" food allergy. Why hay fever patients also react to apples and nuts]. PMID- 11928262 TI - [Depression, divorce, malpractice, bankruptcy. Why do so many physicians commit suicide?]. PMID- 11928264 TI - [Emergencies in general practice. Hemoptysis]. PMID- 11928263 TI - [Steroid sprays in non-infectious rhinitis and sinusitis. Proper and regular spraying does not damage the nasal mucosa]. AB - Glucocorticoids are an effective therapeutic option for most forms of non infectious rhinitis and sinusitis. In Germany, dexamethasone, beclomethasone diproprionate, flunisolide, fluocortin butyl ester, budesonide, fluticasone proprionate, triamcinolone acetonide and mometasone furoate are available for nasal application. The action of these drugs is mediated by cytoplasmic receptors (glucocorticoid receptors). In addition, receptor-independent immediate effects are also known. To ensure optimal efficacy of corticoid sprays, detailed patient instruction is of importance, with particular emphasis being placed on the correct spraying technique and regular application. Modern preparations are associated with very few side effects. There is no evidence that mucosal atrophy occurs. PMID- 11928265 TI - [Do you know what needs to be done here? Frost bite, blood blisters, hypothermic shock]. PMID- 11928266 TI - [Electrolytes in cardiac arrhythmias. Especially effective in coronary heart disease patients]. PMID- 11928267 TI - [Principles in general ultrasound practice. 2: Gallbladder diseases]. PMID- 11928268 TI - [Bluish red skin and abdominal pain in heparin therapy. What ails this elderly woman?]. PMID- 11928269 TI - [Fixed combination of salmeterol and fluticasone. A mile stone in asthma therapy]. PMID- 11928270 TI - [Good asthma control is possible. Synergy of 2 soloists]. PMID- 11928272 TI - [New indication for selective COX-2 inhibitors. Now also approve for acute pain]. PMID- 11928271 TI - [Atypical neuroleptics in dementia. Managing behavioral disorders early and efficiently]. PMID- 11928273 TI - [Therapy of hypertension with ACE inhibitors. One third fewer cases of diabetes]. PMID- 11928274 TI - [Already allergic to pollen in early childhood. Can future asthma still be prevented?]. PMID- 11928275 TI - [Blood pressure amplitude in the morning must be reduced. Saving the hypertensive patient from infarct]. PMID- 11928276 TI - [Suspected bladder carcinoma. New rapid test with high sensitivity]. PMID- 11928277 TI - [Inhibitors of 5-lipoxygenase]. AB - Leukotrienes are powerful mediators of inflammation processes. Possibilities of their biosynthesis inhibition have been intensively studied during recent two decades aiming to develop new methods of therapy of various inflammatory diseases, particularly asthma. Biosynthesis of all leukotrienes is dependent on the production of the key intermediate LTA4. Transformation of arachidonic acid to LTA4 is catalyzed by the enzyme 5-lipoxygenase and facilitated by FLAP. The present paper discusses the results of recent development of new drugs in this area. PMID- 11928278 TI - [The ACE inhibitor, captopril, in the light of new clinical studies]. AB - Captopril, the classic inhibitor of the angiotensin converting enzyme, was employed in several large clinical studies in recent years. The effect of captopril was compared either with placebo, or captopril was selected as the reference ACE inhibitor for comparison with another therapy. In the classic study SAVE, captopril administered to patients after myocardial infarction with a dysfunction of the left chamber reduced mortality by 19%. Though in the study ELITE the AT1 blocker losartan was more effective to reduce mortality in patients with chronic heart failure than captopril, the larger and mortality-oriented study ELITE II did not demonstrate a difference in mortality reduction between captopril and losartan. ACE inhibitors thus remain drugs of choice in chronic heart failure. AT1 blockers are to be used in the cases when ACE inhibitors are not tolerated. The study CAPPP has demonstrated that captopril in hypertonic patients not only effectively decreases blood pressure but exerts a similar effect on mortality reduction as the classic treatment with a diuretic and a betablocker, the most effective being captopril in diabetic patients. Administration of captopril in hypertonic patients with diabetes mellitus in the study UKPDS had an effect on mortality reduction as well as micro- and macrovascular complications of diabetes similar to that of atenolol. The ongoing study VALIANT compares the AT1 blocker valsartan or a combination of valsartan and captopril with captopril alone on patients at risk after myocardial infarction. Also at the beginning of the 21st century captopril maintains a stable position in the treatment of the cardiovascular system. PMID- 11928279 TI - [Infant formulas based on dried milk and risks of its contamination with residual pesticides]. AB - Residues of agricultural pesticides have the potential to enter the organism of farm animals and, subsequently, they can be transferred to milk. Infant formulas are manufactured from dried hydrolyzed cow's milk. Therefore, attention should be paid to possible contamination of the raw material for the manufacture of infant formulas with pesticide residues, especially those of persistent triazine herbicides. Dried milk based formulas are often the only food of young children in their first months of life. Thus, the control of infant milk formulas for pesticide residues is an important issue. PMID- 11928281 TI - [Pharmacoeconomics in the therapy of peptic ulcer]. AB - The great prevalence and chronic nature of peptic ulcers traditionally represent a great economic load for the system of health care. The exponentially growing therapeutic and diagnostic possibilities of the treatment of peptic diseases are under a strong economic pressure from health-insurance companies. The use of an economic model for the management of peptic diseases aims to maximize health benefit from limited sources of the health services. It can be said already now that the detection of the Helicobacter pylori infection ranks among the most important discoveries of the 20th century. Elimination of the Helicobacter pylori infection can produce a dramatic reduction in relapses of peptic ulcers, which is reflected not only in the improved quality of life but also in a subsequent economic contribution. The present authors paid attention to the financial costs of the combined therapy for eradication of the Helicobacter pylori infection in the course of the years 1998-2001. PMID- 11928280 TI - [Effect of Consupren, Carvedilol and BL-443 on the composition of lactate dehydrogenase in tissues of rats with cyclosporin nephropathy]. AB - L-Lactate dehydrogenase pattern in tissues was determined by polyacrylamide gel electrophoresis adjusted to be more sensitive to LD1 and LD2. Three groups of rats (Wistar) were treated for 18 days with single oral daily doses of 45 mg consuprene/kg body weight to induce cyclosporine nephropathy. Two treated groups were further medicated either with carvedilol or BL-443 in single daily doses of 10 mg/kg b.w., unmedicated rats were given single i.p. daily doses of 1 ml saline. No significant difference in the LD(1-4) isoenzyme pattern in the liver between intact rats, rats with cyclosporine nephropathy, and rats with cyclosporine nephropathy medicated with carvedilol or BL-443 was found by F-test and t-test (p < 0.05). However, a significant difference in the LD(1-4) isoenzyme pattern in the myocardium between rats with cyclosporine nephropathy and intact rats was found. The present study reports the preliminary results of the effects of consupren on the LD(1-4) pattern in the muscle, spleen, and lung, as well as the effects of carvedilol and BL-443 in the tissues under the conditions of experimental cyclosporine nephropathy. The tissues with an increased risk of affection can be identified by evaluation of LD patterns that may become an additional tool to microscopic examination of the sample. PMID- 11928282 TI - [Reperfusion injury in the isolated rat liver after hypothermic preservation]. AB - Histological changes which appear as a result of reperfusion injury of cold preserved rat liver were studied at intervals of 0 hr, 3 hr, 24 hr and 48 hr of cold storage. The isolated livers were stored in a UW solution (University of Wisconsin), which is used in human liver transplantations. Computer image analysis of light microscopic sections (methyl green-pyronin stained) was used for the study and quantification of injured cells. The method of TUNEL was performed to prove possible apoptosis of sinusoidal endothelial cells and heptocytes. Bile production during reperfusion and ALT, AST, LDH and ACP were measured in the reperfusion medium at the end of the 90 min reperfusion. It has been confirmed that prolongation of the cold storage of liver results in extensive changes in the liver structure and increased injury of liver cells. Sinusoidal endothelial cells were damaged more and earlier than hepatocytes. It has been shown that methyl green-pyronin stained sections are advantageous for the study of these morphological changes, allowing the strongest view of these changes. The appearance of TUNEL positive cells and an increase in the levels of biochemical parameters, e.g. AST or ALT, indicate earlier cell injury. The methodology described in this article can be used for the study of reperfusion injury of the liver and for the study of this phenomenon in other experiments. PMID- 11928283 TI - [Validation of the HPLC method in the determination of dioxopromethazine and phenylephrine in eye drops]. AB - The present paper introduces a rapid HPLC method for the determination of dioxopromethazine and phenylephrine in eye drops. The method uses a modified C18 stationary phase optimized for the separation of basic compounds and a methanol/1.5 mM phosphoric acid (60/40 v/v, pH 3.02) mobile phase. The flow rate is set to 2 ml/min, sample volume 20 microliters, and compounds are detected at 275 nm. Prior to analysis, the eye drops are diluted with water in a ratio of 1:50. The elaborated HPLC method and the chromatographic system were validated according to the procedure for the validation of chromatographic systems and methods. PMID- 11928284 TI - [Effect of jasminic acid on production of flavonoids in a culture of Ononis arvensis L. in vitro]. AB - The present paper examined the effect of jasminic acid in three different concentrations (1000 mg/l; 10 mg/l, and 0.1 mg/l) on the formation of flavonoids in the culture Ononis arvensis L. in vitro. The tested concentrations of jasminic acid were applied to the callus and suspension cultures of Ononis arvensis L. in the 4th week of culture growth and elicitation took place for 12; 24; 48; 72, and 168 hours. A statistically significant increase in the content of flavonoids was found in all administered solutions of jasminic acid after 12 hours of elicitation, excepting the concentration c2 (10 mg/l) in the suspension culture. The highest yield was a 55% increase in the content of flavonoids after elicitation with jasminic acid in the concentration c1 (1000 mg/l) in the callus culture. PMID- 11928285 TI - [Preformulation study of a potential local anesthetic from a group of phenylcarbamic acid derivatives]. AB - Within pre-formulation studies of phenylcarbamic acid derivatives with local anaesthetic effects, potential drug IV with the chemical name N-(2-(2 ethylphenylcarbamoyloxy)-ethyl)-piperidiniumchloride was examined. The study evaluated the effect of polyhydric alcohols--glycerol, propylene glycol, sorbitol and their concentrations of 5; 10; 15, and 20 wt % on the values of the partition coefficient in the system n-octanol/water, surface tension, and viscosity of the solutions of potential drug IV. The optimal auxiliary substance seems to be glycerol of a concentration of 15-20 wt %, as it does not affect the values of the partition coefficient and surface tension of potential drug IV solutions in a statistically significant manner. PMID- 11928286 TI - An influenza prevention campaign: the employee perspective. AB - While the benefits of vaccinating health care workers against influenza are known, the number of vaccinated employees remains inadequate. This article describes survey results from 999 Advocate employees. Statistically significant differences were identified in receiving the influenza vaccine based on age, recommendation of the flu vaccine to others, working environment, and race. The major reason identified for receiving the vaccine was to stay healthy. The most common reason cited for not receiving the vaccine was receiving the vaccine before and getting sick anyway. Results of the study were used to refine education and communication strategies, improving the employee vaccination program. PMID- 11928287 TI - Spanish validation of an instrument to measure the quality of nursing care in hospital emergency units. AB - The article describes the translation and adaptation to Spanish of the Consumer Emergency Care Satisfaction Scale (CECSS), an instrument used to measure the quality of care nurses give in emergency units. Bilingual people using a translation-retrotranslation process performed the adaptation. The reliability and validity of content and construction were analyzed. The reliability study showed satisfactory indexes in all cases. Content validity (judgment of expert nurses and patients) obtained a mean item-dimension suitability valuation of 6.1 and item-scale suitability of 5.5. As the Spanish version of the CECSS is comprehensible to patients, and reliability and validity are satisfactory, it provides a useful measure of the quality of nursing care. PMID- 11928288 TI - Wide application of CQI in home care. AB - Organizations are often overwhelmed when faced with a need to develop a comprehensive quality improvement program and use a cross-organizational team to improve outcomes. This article describes the total quality management program implemented in a long-term home health care program in New York State. It describes common issues, such as incidents, multidisciplinary clinical record documentation audits, and staff competency. As a result of this comprehensive performance improvement process, this organization has not only changed the manner in which it operates but also improved its performance. PMID- 11928289 TI - How can the public identify an excellent healthcare organization? PMID- 11928290 TI - Acute shortage plays out in nursing stations, hospital wards. PMID- 11928291 TI - Drugged and sexually assaulted. PMID- 11928292 TI - Spiritual nursing interventions. PMID- 11928293 TI - Postpartum depression support. PMID- 11928294 TI - Teen depression. A nurse's reflection. PMID- 11928295 TI - A call to focus our "passion for substance" on family violence. PMID- 11928297 TI - Making connections: a vehicle for developing a nursing response to violence against women and children in Canada. PMID- 11928296 TI - Research on violence and abuse in Canada: challenges and opportunities. PMID- 11928299 TI - Portraits of pain and promise: a photographic study of Bosnian youth. AB - In the early 1990s, war erupted in Bosnia and Herzegovina, forcing large numbers of people to flee their homes and country, abandoning their culture and all that was familiar to them. For the children, often described as war's "innocent victims," the conflict and subsequent uprooting represented a dramatic end to their peaceful lives. Although many were fortunate enough to escape with their families and resettle amid more peaceful circumstances, there is considerable evidence that refugee youth are forever changed by their exposure to war and that the pain of war does not end when the fighting is over. This paper presents the results of a study with 7 Bosnian children, aged 11-14, who came to Canada as refugees during the 1990s. The everyday challenges and struggles faced by this group were explored using an innovative research method called photo novella. A secondary purpose of the research was to evaluate the merits and limitations of photo novella as a method for capturing children's perspectives and feelings. Participants were given disposable cameras and asked to take pictures of important people, places, and events. The meaning of the photographs was then explored through a dialogic process the researchers call phototalk. The findings revealed that while these children had many strengths, they continued to struggle to understand the events that so profoundly changed their lives. The results and the implications for nurses are discussed. PMID- 11928298 TI - Development and psychometric testing of the Wilmoth Sexual Behaviors Questionnaire-Female. AB - The purpose of this study was to develop and test the Wilmoth Sexual Behaviors Questionnaire-Female. As initially developed, the WSBQ-F consisted of 54 items arranged in 8 subscales. Items were rated on a Likert-type scale, with high scores reflecting more consistent use of a sexual behaviour. A convenience sample of 310 women with breast cancer (n = 165) and healthy women (n = 145) participated in the study. An Index of Content Validity of 1.00 was obtained. Construct validity was estimated using exploratory factor analysis and the known groups method. Internal consistency reliability was .94 for the total scale, with coefficients ranging between 0.52 and 0.94 for the subscales. The test-retest correlation coefficient was .81 for the total scale; coefficients were between .58 and .88 for the subscales. Initial testing of the measure suggests that the WSBQ-F is a reliable and valid measure of female sexual behaviours. PMID- 11928300 TI - A celebration of nursing research on violence. PMID- 11928301 TI - Battering and breastfeeding in a WIC population. AB - The study reported in this paper was based on the hypothesis that women who are victims of domestic violence may be less likely to select breastfeeding than women who are not abused. Informed consent was obtained from 212 women at 2 Women, Infants and Children (WIC) Nutritional Supplemental Program clinics in the Midwestern United States. The Abuse Assessment Screen was administered by interview and women were also questioned about intended feeding choice and whether they had breastfed any previous children. No association was found between present or previous abuse and infant-feeding choice. Nevertheless, the findings of this study can be considered important, for two reasons: (1) this was an initial inquiry examining the relationship between having been abused and ability to choose the feeding method of a newborn; and (2) women in the sample who reported present or current abuse were able to breastfeed their infants in the same proportion as those who did not report abuse, which suggests that a woman's concern for her child overcomes her possible fears of control by the batterer. PMID- 11928302 TI - Nurses' experience of violence in Alberta and British Columbia hospitals. AB - This study examined responses to a survey on violence in the workplace from a sample of 8,780 registered nurses practising in 210 hospitals in the Canadian provinces of Alberta and British Columbia. Findings relate to the frequency of violence against nurses, reported as the number of times they experienced a violent incident in the workplace. Nearly half (46%) of those surveyed had experienced 1 or more types of violence in the last 5 shifts worked. Frequency varied by type: emotional abuse 38%, threat of assault 19%, physical assault 18%, verbal sexual harassment 7.6%, sexual assault 0.6%. Further, 70% of those who had experienced violence indicated they had not reported it. Patients constituted the main source of all types of violence. The most prevalent type, emotional abuse, was further explored for its possible determinants. This was also the type of violence most evenly distributed among sources (patients, families, co-workers, physicians). Multiple regression modelling using the individual nurse as the unit of analysis showed the significant predictors of emotional abuse to be age, casual job status, quality of care, degree of hospital restructuring, type of unit, relationships among hospital staff, nurse-to-patient ratios, and violence prevention measures; using the hospital as the unit of analysis the predictors were found to be quality of care, age, relationships with hospital staff, presence of violence-prevention measures, and province. These findings illustrate important differences in models that use the individual and the institution as the unit of analysis. Implications include targeting prevention strategies not only at the nurse but, perhaps more importantly, at the hospital. Overall, the findings suggest that health-care institutions are not always healthy workplaces and may increasingly be stressful and hazardous ones. PMID- 11928303 TI - Beyond survival: reclaiming self after leaving an abusive male partner. AB - Research on the process of leaving an abusive male partner has focused on surviving abuse and the crisis of leaving. Little is known about the experience of women who have left abusive male partners and not gone back. In this feminist grounded theory study of women leaving abusive partners, the researchers discovered the basic social-psychological process of reclaiming self in which women voyaged through 4 stages: counteracting abuse, breaking free, not going back, and moving on. The focus of this paper is the last stage, moving on, during which women move beyond framing their lives as survivors of an abusive relationship through the processes of figuring it out, putting it in its rightful place, launching new relationships, and taking on a new image. The findings extend our knowledge of the leaving process by delineating the ways in which the abuse experience and the survival process are displaced as the centre of the woman's intra-psychic, interpersonal, and social existence. Questions are raised about how nurses and other health professionals can avoid revictimizing women who have moved on. PMID- 11928304 TI - Abuse obscured: an ethnographic account of emergency nursing in relation to violence against women. AB - Violence against women is a significant health issue in Canada and around the globe, yet the health-care response has been inadequate. While various reasons for this inadequacy have been suggested, little systematic research has been undertaken. This ethnographic study of 2 hospital emergency units was conducted to describe nursing practice in relation to violence against women. Participant observation and interviews with 25 healthcare providers and 5 patients in the 2 units were complemented by interviews with 5 nurses from other emergency units. The findings illustrate that abuse is obscured and practice shaped by stereotypical thinking and a focus on physical problems and rapid patient processing. Perceptions of patient deservedness influenced care that ranged from "doing nothing" to actively offering the patient choices. This description provides a basis for designing meaningful education for nurses and systemic changes that will foster more effective practice. PMID- 11928305 TI - [Total patient care as a goal in geriatric psychiatry]. PMID- 11928306 TI - [Guidelines for care of patients with mental disorders]. PMID- 11928307 TI - [Guidelines for the care of patients with mental disorders--link between community mental health and patient care management]. PMID- 11928308 TI - [Promotion of patient care management for mental disorders and the ideal application]. PMID- 11928309 TI - [Empowerment and advocacy as goals in geriatric psychiatry]. PMID- 11928310 TI - [Client-driven model of community total care management for patients with disabilities ]. PMID- 11928311 TI - [Future aspects for patient care management of mental disorders in Japan]. PMID- 11928312 TI - [Care guidelines for patients with mental disorders in psychiatric hospitals]. PMID- 11928313 TI - [Attention deficit hyperactivity disorder]. PMID- 11928314 TI - [Basic and clinical studies on solvent abuse]. PMID- 11928315 TI - [Pharmacogenetic studies on psychotropic agents]. PMID- 11928316 TI - [Current status of studies of schizophrenia after DSM-III]. AB - The reinforcement of biological studies of schizophrenia by the DSM-III had been promoted by the general medicalization of psychiatry itself, following the successful introduction of antipsychotic drugs as well as the advance in brain research. The traditional view of schizophrenia has included since the time of Kraepelin till present the presumed biological basic diseases and specific psychological reaction. The DSM notion of schizophrenia presumes exclusively biological aspects in that it is based on a very simple unitary model, although the lack of sufficient validity has not been compensated by the advance in the biological studies that produced a number of related findings but not the marker of schizophrenia. The exclusion of the concept of psychosis makes it difficult to discuss the semantic isolation in which the patients of this disorder are often entrapped. The symptoms are regarded as indices of presumed biological diseases rather than a product of semantic interaction. The simple model of schizophrenia is easy to use in biological studies but too weak to give clinical significance to the various findings detected. It will be insightful to look into other modes of diagnosis such as Kleist-Leonhardt or French school as well as subtle basic disturbance in German school that are not mentioned in the current DSM system. PMID- 11928317 TI - [Result of urgent investigation into the actual condition of serious criminal patients hospitalized after psychiatric evaluation in conformity with the mental health welfare law]. PMID- 11928318 TI - [Total patient care and welfare measures as goals in geriatric psychiatry]. PMID- 11928319 TI - [Epidemiological studies on the etiology of biliary tract cancers]. PMID- 11928320 TI - [Elucidation of toxicity mechanism of environmental chemicals induced by oxidative stress]. PMID- 11928321 TI - [Elucidation of paraquat poisoning mechanism and development of the neuronal death model]. PMID- 11928322 TI - From rhetoric to reality. PMID- 11928324 TI - The psychosocial impact of incontinence on older people: a review. PMID- 11928323 TI - Readmissions of older people to acute medical units. PMID- 11928325 TI - Standing and moving: helping people with vascular dementia. AB - Safe handling and movement of older people requires an understanding among nurses and carers of the patterns of normal movement--for example, from sitting to standing. But there are extra potential difficulties when helping people with vascular dementia. This article offers guidance on how those problems may be overcome. PMID- 11928326 TI - Identifying and assessing abuse in care homes--Part 2. PMID- 11928327 TI - Unmet need means pressures mount. PMID- 11928328 TI - Artificial feeding. PMID- 11928329 TI - Longer shift, better care? PMID- 11928330 TI - Pressure sore prevention. PMID- 11928331 TI - Faith and hope in charity. PMID- 11928332 TI - Critical theory: critical methodology to disciplinary foundations in nursing. AB - Increasingly in the nursing literature, theorists have examined the use of critical theory in nursing (especially as understood by Habermas) and many have advocated it as a research approach to guide knowledge development in nursing. There has been limited analysis, however, of critical theory's broader foundational implications for the discipline of nursing. Part of the difficulty stems from a failure to differentiate between the implications of Habermas's earlier work on knowledge interests and his later theory of communicative action. In this paper, Habermas's critical theory is explored along two dimensions: as a metatheoretical account of a methodology of critical theory as a research tradition; and as a theory of communicative action whose dialogical and normative assumptions have profound implications for a postfoundationalist grounding of nursing as a discipline and professional practice. The authors argue that critical theory is necessary for nursing and may be sufficient as a paradigmatic philosophical base for the discipline. PMID- 11928333 TI - Home care in Canada: passing the buck. AB - This paper provides an overview of Canadian home-care utilization, highlights the health-policy assumptions that have resulted in an increasing reliance on in-home services, and assesses the current roles of the private and public sectors in the financing of home care. Significant interprovincial variations in per capita home care expenditures and potential inequalities in access to home care call for resolution by federal and provincial governments. There is a need for consensus with respect to medically and socially necessary services that are subject to national standards, irrespective of the setting in which services are sought, received, and delivered. The development and enforcement of national home-care standards that complement the principles of the Canada Health Act would be a useful first step in ensuring that the Canadian health-care system is ready to confront the challenges of the new millennium. PMID- 11928334 TI - The material and social predicaments of home: women's experiences after aortocoronary bypass surgery. AB - Several authors argue that women's lives are conditioned by social locations such as class, race, ethnicity, age, and chronic illness or (dis)ability. Patterns of advantage and disadvantage, domination and oppression are formed which constitute the groundwork of women's health. An institutional ethnography was designed to follow the experiences of 18 women on their return home following aortocoronary bypass surgery. Using the narratives of 3 women as examples, the author highlights the everyday activities pursued by the women in the first month after their return home. The author describes the circumstances under which the activities occurred and discusses the social relations reflected in the descriptions. From this analysis it is argued that research and substantive work would benefit from a more critical understanding of women's different experiences of the home and of home care. PMID- 11928335 TI - Perceived support needs of family caregivers and implications for a telephone support service. AB - The purposes of this study were: to identify the perceived support needs of family caregivers of persons living with chronic illness (physical or cognitive) and receiving home-care services, and to describe the types of telephone services that would meet the expressed needs of caregivers. The qualitative design used semi-structured interviews. A total of 34 caregivers (mean age 62 years) participated in the study. The care recipients (mean age 78 years) were primarily the husband/wife or parent of the caregiver. The most commonly expressed caregiver needs were: a social life, instrumental support (e.g., respite, assistance with physical care, financial compensation), informational support, and emotional support. Most caregivers said they would use a telephone support service provided by a professional (71%) or a fellow caregiver (59%) if available. The results of this study support a pilot study and evaluation of a telephone support service for family caregivers. PMID- 11928336 TI - The politics of home care: where is "home"? PMID- 11928337 TI - Negotiating care of frail elders: relationships between community nurses and family caregivers. AB - Recent changes in patterns of care of the elderly in Canada, including the withdrawal of formal home-care services and increasing reliance on family caregivers, call for a critical analysis of the relationship between formal and informal caregivers. The purpose of this study was to describe and analyze the relationship between nurses and female family members caring for frail elders in the home. Using a critical ethnographic method in a socialist-feminist framework, separate in-depth interviews were conducted with 23 nurse-family caregiver dyads. Analysis of interview transcripts and fieldnotes revealed that relationships were characterized by uncertainty and tension. While both nurses and family caregivers functioned within and resisted current home-care arrangements, they engaged in an ongoing process of negotiating cultural assumptions about "private" and "public" caregiving. The findings point to implications for nursing practice, education, and policy. PMID- 11928338 TI - Centring the home in research. PMID- 11928339 TI - Making evidence-based practice a reality: The National Center for Children, Families and Communities. PMID- 11928340 TI - Stress and the effects of hospital restructuring in nurses. AB - This study examines the extent of stress and burnout experienced by nurses during hospital restructuring. It includes both job-related outcomes such as job satisfaction and burnout, and psychosomatic outcomes such as depression. The study compares effects attributable to number of hospital restructuring initiatives with those attributable to specific work stressors such as workload, bumping (where one nurse replaces another due to greater seniority), and use of unlicensed personnel to do the work of nurses. It also examines the role of personal resources including self-efficacy and coping. Results show that, in hospitals undergoing restructuring, workload is the most significant and consistent predictor of distress in nurses, as manifested in lower job satisfaction, professional efficacy, and job security. Greater workload also contributed to depression, cynicism, and anxiety. The practice of bumping contributed to job insecurity, depression, and anxiety. The results point to specific deleterious effects of hospital restructuring. Implications of the findings are discussed. The extent to which workload issues are managed through appropriate practices can be expected to match the extent of nurses' experience of either job satisfaction or depression and anxiety. Such practices need to be part of an ongoing process of interaction between the hospital administration and nurses. PMID- 11928341 TI - The aetiology of gingival recession. AB - Gingival recession affects a large proportion of the adult population. This paper discusses the aetiology and mechanisms behind the formation of gingival recession, and considers the relevance of gingival recession to both orthodontics and restorative dentistry. It is followed by a second paper on the management of gingival recession. PMID- 11928343 TI - The management of blood phobia and a hypersensitive gag reflex by hypnotherapy: a case report. AB - Coping with a hypersensitive gag reflex can be a cause for concern for both the patient and the operator. This report describes a case of blood phobia directed solely towards the oral cavity, linked with the inability to tolerate dentures due to a hypersensitive gag reflex. Management by hypnotherapy using a systematic desensitization technique allowed for extraction of teeth and permanent elimination of the gagging problem. PMID- 11928342 TI - Preserving the vital pulp in operative dentistry: I. A biological approach. AB - This is the first in a series of four papers aimed at understanding human pulpal responses to tissue injury, cavity preparation and restorative events. This article provides an insight into the exquisite regenerative potential of the dentine-pulp complex which underpins the success of restorative dentistry. PMID- 11928344 TI - Burning mouth syndrome and vulvodynia coexisting in the same patient: a case report. AB - The 'dynias' are a group of chronic focal pain syndromes with a predilection for the orocervical and urogenital regions. This is a case report of stomatodynia (burning mouth syndrome) and vulvodynia coexisting in a middle-aged woman. The dynias are an enigma in terms of aetiology, which is multifactorial, making clinical investigations difficult and often requiring liaison with other specialties. PMID- 11928345 TI - Double fracture and wedging of a sewing needle interdentally in an attempt at self-treatment: a case report. AB - Dental pain occurring whilst abroad may present the patient with a difficult decision: to try and bear it with self-medication until he/she returns home or to seek professional help. This case report outlines a Korean girl's attempt at self treatment, which resulted in not one but two fragments of a sewing needle wedging between her teeth, the second fracturing as she tried to dislodge the first. Her self-treatment exacerbated her problem and resulted in the need for surgical access to the site to retrieve the broken fragments. The case also illustrates the use of parallax radiography in determining the position of the fragments. PMID- 11928346 TI - Research in dental practice: a 'SWOT' analysis. AB - Most dental treatment, in most countries, is carried out in general dental practice. There is therefore a potential wealth of research material, although clinical evaluations have generally been carried out on hospital-based patients. Many types of research, such as clinical evaluations and assessments of new materials, may be appropriate to dental practice. Principal problems are that dental practices are established to treat patients efficiently and to provide an income for the staff of the practice. Time spent on research therefore cannot be used for patient treatment, so there are cost implications. Critics of practice based research have commented on the lack of calibration of operative diagnoses and other variables; however, this variability is the stuff of dental practice, the real-world situation. Many of the difficulties in carrying out research in dental practice may be overcome. For the enlightened, it may be possible to turn observations based on the volume of treatment carried out in practice into robust, clinically related and relevant research projects based in the real world of dental practice. PMID- 11928347 TI - Trismus: aetiology, differential diagnosis and treatment. AB - Trismus is a problem commonly encountered by the dental practitioner. It has a number of potential causes, and its treatment will depend on the cause. This article discusses the primary causes of this condition and the various treatments available. PMID- 11928348 TI - The report of the Information Technology Task Group of the Dentistry Modernization Steering Group. AB - The Dentistry Modernization Steering Group was set up to monitor, evaluate and facilitate progress in the implementation of the Government's strategy for the dental services. Information Technology was an important part of this remit and the report of its IT Task Group is summarized here. PMID- 11928349 TI - Treatment planning for the loss of first permanent molars. PMID- 11928350 TI - Successful restoration of load-bearing cavities. PMID- 11928351 TI - Japanese carbon storage in materials. AB - The NEAT model (Nonenergy-use Emission Accounting Tables) has been developed in order to estimate CO2 emissions caused by so-called nonenergy use of fossil fuels. The model is based on material flow accounting. The model has been applied to a number of countries in order to validate and improve its use. This paper discusses the case study for Japan. The NEAT analysis suggests that emissions in 1996 were 23 Mt higher than previously estimated based on the guidelines of the Intergovernmental Panel on Climate Change (IPCC). This quantity equals 1.9% of the total Japanese greenhouse gas emission. It is recommended to adjust the Japanese emission accounting practice and to apply more detailed emission estimation methods in future years. Given similar results for other countries it is also recommended to improve the IPCC guidelines. PMID- 11928352 TI - Sustainable watershed management: an international multi-watershed case study. AB - Global freshwater resources are being increasingly polluted and depleted, threatening sustainable development and human and ecosystem health. Utilizing case studies from 4 different watersheds in the United States, Japan, Switzerland, and Brazil, this paper identifies the most relevant sustainability deficits and derives general vectors for more sustainable water management. As a consequence of the demographic and economic developments experienced in the last few decades, each watershed has suffered declines in water quality, streamflow and biotic resources. However, the extent and the cultural perception of these water-related problems vary substantially in the different watersheds, leading to specific water-management strategies. In industrialized countries, exemplified by the US, Switzerland, and Japan, these strategies have primarily consisted of finance- and energy-intensive technologies, allowing these countries to meet water requirements while minimizing human health risks. But, from a sustainability point of view, such strategies, relying on limited natural resources, are not long-term solutions. For newly industrialized countries such as Brazil, expensive technologies for water management are often not economically feasible, thus limiting the extent to which newly industrialized and developing countries can utilize the expertise offered by the industrialized world. Sustainable water management has to be achieved by a common learning process involving industrialized, newly industrialized, and developing countries, following general sustainability guidelines as exemplified in this paper. PMID- 11928353 TI - Ozone dynamics and deposition processes at a deforested site in the Amazon basin. AB - The goal of this study is to investigate the impact of deforestation on ozone dynamics and deposition in the Brazilian Amazon basin. This goal is accomplished through i) analyses of ozone levels and deposition rates at a deforested site during the rainy season; and ii) comparisons of these data with similar information derived at a forest. At the pasture site maximum ozone mixing ratios reach 20 parts per billion on a volume basis (ppbv) but about 6 ppbv prevail over the forest. Maximum ozone deposition velocities for pastures can reach 0.7 cm s 1, which is about threefold lower than values derived for forests. Combining ozone abundance and deposition velocities, pasture maximum ozone fluxes reach approximately 0.2 microgram (ozone) m-2 s-1. This flux represents approximately 70% of the deposition rates measured over the forest. Hence, this study suggests that conversion of rainforests to pastures could lead to a net reduction (30%) in the ozone sink in the Amazon. PMID- 11928354 TI - Links between Cairo and Kyoto: addressing global warming through voluntary family planning. AB - Over the past three decades, with a combination of new technology, rising female literacy rates, and strengthened family planning programs, the world has seen dramatic increases in the use of contraception, with corresponding declines in fertility and population growth rates. At the International Conference on Population and Development, Cairo in 1994, parties pledged a tripling of funding for reproductive health programs in developing countries. Many demographers believe that making such programs more widely available to women would extend the decline in birth rates and shift the world towards the low scenario of United Nations population projections over the next century and a half. By examining the costs and impacts of such programs, in view of the links between population and carbon emissions, this paper shows that extension of voluntary family planning could make a large and cost-effective contribution to the greenhouse gas limitation goals of the Kyoto Protocol that was negotiated in 1997. PMID- 11928355 TI - Constraints to implementing international agreements: the case of the Montreal Protocol in Botswana. AB - This study addresses the difficulties surrounding effective implementation of the Montreal Protocol in Botswana and provides a general understanding of how best we might advise policy makers when implementing international agreements in the developing world. A questionnaire survey administered to both the formal and informal users of Ozone Depleting Substances (ODSs) revealed that disseminated information on ODSs has little effect on choices that users make about refrigerant gases and this information is skewed in favor of the conventional users of ODSs. As a result, annual statistics of ODS use are probably underestimated. Difficulties exist in changing from old to new technologies in the short term due to high costs associated with the change over. The infrastructure to recover gases and to dispose of unusable hardware is absent or inadequate. Solutions to these difficulties include a comprehensive policy that caters for all users of ODSs and the integration of economic and environmental aspirations. PMID- 11928356 TI - Renewable energies in electricity generation for reduction of greenhouse gases in Mexico 2025. AB - This study presents 4 scenarios relating to the environmental futures of electricity generation in Mexico up to the year 2025. The first scenario emphasizes the use of oil products, particularly fuel oil, and represents the historic path of Mexico's energy policy. The second scenario prioritizes the use of natural gas, reflecting the energy consumption pattern that arose in the mid 1990s as a result of reforms in the energy sector. In the third scenario, the high participation of renewable sources of energy is considered feasible from a technical and economic point of view. The fourth scenario takes into account the present- and medium-term use of natural-gas technologies that the energy reform has produced, but after 2007 a high and feasible participation of renewable sources of energy is considered. The 4 scenarios are evaluated up to the year 2025 in terms of greenhouse gases (GHG) and acid rain precursor gases (ARPG). PMID- 11928357 TI - Erosion vs. recovery of coral reefs after 1998 El Nino: Chagos reefs, Indian Ocean. AB - Three years after most corals died on the central Indian Ocean reefs of Chagos, erosion and recovery were studied to 30 m depth. Mortality was near-total to 15 m deep in northern atolls, and to > 35 m in central and southern atolls. Some reef surfaces have 'dropped' 1.5 m due to the loss of dense coral thickets. Coral bioerosion is substantial, reducing 3-D reef 'structure' and forming unconsolidated rubble. Juvenile corals are abundant, though mostly on eroding or unstable substrates, and are of less robust species. Reef fish abundance and diversity at 15 m depth remains high; species dependent on corals have diminished, while some herbivores and detritivores have increased. A new sea surface temperature (SST) data set shows that mean SST has risen 0.65 degree C since 1950. The critical SST causing the mortality in Chagos was 29.9 degrees C. PMID- 11928358 TI - Local hunting and the conservation of large mammals in India. AB - Hunting by local communities is among the most widespread threats to Indian wildlife, yet, the understanding of its nature, extent, and impacts on wildlife has been poor. We surveyed 2 protected areas--Kudremukha and Nagara-hole--in southern India to assess the impacts of local hunting on large mammals. Detailed interviews with retired and active hunters were employed to describe hunting patterns. Impacts of hunting were assessed by comparing large-mammal abundance in adjacent sites differing in their vulnerability to hunting. In Kudremukha, at least 26 species of mammals were hunted, mostly with guns, at an estimated intensity of 216 hunter-days per month per village. In Nagarahole, 6 of the 9 focal species of large mammals occurred at significantly lower densities at the heavily hunted site where enforcement capabilities were poorer. Our data underscore the importance of preservationist programs in the conservation of large mammals in a context of extensive local hunting. PMID- 11928359 TI - State and school activities. PMID- 11928360 TI - Learning lifelong leadership skills through NSNA. PMID- 11928362 TI - Challenges of the nursing shortage. Implications for the future. PMID- 11928363 TI - Health care organizations. Respond to shortage. PMID- 11928361 TI - A research internship. Offers life lesson. PMID- 11928365 TI - The nursing shortage.... What nursing students can do. PMID- 11928364 TI - What New York is doing about the nursing shortage. PMID- 11928366 TI - The image of nursing. Past, present, and future. PMID- 11928367 TI - Hailing one of health care's priceless resources--nurses. PMID- 11928368 TI - [Advances in geriatrics]. PMID- 11928370 TI - [Thromboembolic diseases. XVIIIth Congress of the International Society on Thrombosis and Haemostasis, Paris, 6-12 July 2001]. PMID- 11928372 TI - [Can we still speak of Wissler-Fanconi syndrome?]. PMID- 11928373 TI - [Non-aggressive diagnostic approach to invasive pulmonary aspergillosis in a hematology unit. Retrospective analysis of a series of 16 cases]. AB - PURPOSE: Among neutropenic patients, diagnosis of invasive pulmonary aspergillosis is difficult. Computed tomographic scan, bronchoalveolar lavage and histology are considered invasive procedures, because they represent an infectious risk for these immunocompromised patients. METHODS: We describe the clinical and noninvasive paraclinical (X-rays, serology) signs of invasive pulmonary aspergillosis, from a retrospective study of 16 cases in a haematology unit. RESULTS: Outside of fever and chills, cough and polypnea are the earliest signs, followed by chest pain, dyspnea, lung auscultation changes, and haemoptysis. The sensitivity of each sign is higher than 56%. Before the onset of lung auscultation changes, the chest X-ray shows mainly unilateral alveolar infiltrates. Sensitivity of serology is weak (25%), but contributed to early diagnosis in 16.6% of cases. CONCLUSION: A better knowledge of the invasive pulmonary aspergillosis clinical, radiological and serological signs could help the practician to prescribe an 'invasive' investigation (computed tomographic scan, bronchoalveolar lavage) to confirm the diagnosis of this fungal infection. PMID- 11928374 TI - [Vascular complications of homocystinuria: a retrospective multicenter study]. AB - PURPOSE: Arterial or venous thromboses are frequent in patients with homocystinuria. Because severe homocystinuria is rare, prevalence of thrombosis, especially in France, is still unknown. METHODS: Review of the clinical outcome of 37 patients with homocystinuria due to cystathionine-cystathionine beta synthase deficiency (34) and 5,10-methylenetetrahydrofolate reductase (three) lead us to describe vascular complications occurring in 12 (32%) of them. RESULTS: Venous thromboembolism is the earlier and the most frequent one and is mainly found in untreated late-diagnosed cases. Under specific treatment of homocystinuria, thromboses are rare and always a complication of surgery associated with high thromboembolic risk. Association with factor V Leiden increased the risk of venous thrombosis. PMID- 11928376 TI - [Hepatitis C and pregnancy]. AB - PURPOSE: Today, the natural course of hepatitis C virus (HCV) infection during pregnancy and the prevalence of mother-to-child transmission are better known. CURRENT KNOWLEDGE AND KEY POINTS: Antenatal screening for HCV infection needs to be proposed to women with risk factors. Viral replication needs to be confirmed by PCR in pregnant women with antibodies against HCV. To date, the clinical course of pregnancy and the mode of delivery have not been changed by HCV infection. Rates of vertical transmission of HCV are about 6% in women with HCV alone and 15% in women co-infected with HIV. A screening for HCV markers is required 18 months after delivery for infants born to HCV mothers. Because of the relatively low rate of HCV vertical transmission, pregnancy can be allowed in infected women. However, taking into account the efficacy of new antiviral strategies, treatment of HCV infection could be proposed before pregnancy. FUTURE PROSPECTS AND PROJECTS: In case of HCV infection, a careful follow-up of both mother and newborns is required. Long-term follow-up of infected infants is needed to assess the consequences of perinatal HCV infection. PMID- 11928375 TI - [The antisynthetase syndrome: a subgroup of inflammatory myopathies not to be unrecognized]. AB - PURPOSE: Antisynthetase syndrome (AS) is frequently revealed by interstitial lung disease and arthritis. There are mechanic's hand, Raynaud's phenomenon and anti aminoacyl t-RNA synthetase antibodies. The anti JO-1 antibody is the most frequently identified. We report five cases of antisynthetase syndrome with particular clinical features and good response to corticosteroids. METHODS: There are three women and two men with a median age of 59 years at presentation (range: 44-77). Three patients progressively developed AS: the symptoms are dyspnea (three). Raynaud's phenomenon (one), purpura (one) and hyperkeratosis, scaling and fissuring on the lateral sides of the fingers (two). Patients always had skin signs: hyperkeratosis and scaling (five), purpura (one), Raynaud's phenomenon with normal capillaroscopy (two). Lung disease is present in the five cases with interstitial lesions in CT scans (five), trouble of CO diffusion (three/three) and lymphocytic alveolitis (two/two). Moderate muscular disorders are present in five cases (moderate elevated muscular enzyme: five, positive muscle histology: two). Anti-JO-1 antibodies are present in five cases. AS is associated with connective tissue diseases: rheumatoid polyarthritis in one case and Gougerot Sjogren in three cases. No malignant tumour is associated. Patients have received oral corticosteroid treatment (five/five) with high doses of intravenous perfusions (three/five) with, initially, a good response. For only one patient, immunosuppressive treatment was necessary because of the articular relapse. The interstitial lung disease had a good response to corticosteroids therapy alone in four cases. Because of the relapse during the tapering off of corticosteroids, corticosteroids were increased in one case and immunosuppressive therapy was required in one case. CONCLUSION: The prognosis of AS depends of the interstitial lung disease. High doses of corticosteroids are required. In our study, the response to corticosteroids is good. Immunosuppressive agents must be added in severe and progressive form of interstitial lung disease in AS. PMID- 11928377 TI - [Idiopathic sudden deafness]. AB - PURPOSE: Sudden idiopathic deafness is a sensorineural hearing loss with no recognized causes at the time of onset. The impairment site is usually localized in the cochlea, but some cases of retrocochlear lesions (e.g., cerebellopontine angle tumors, degenerative neural diseases, neuraxial ischemic lesions) can induce sensorineural deafness. The medical management of patients presenting with sudden deafness aims at detecting a causal mechanism, and at administering emergency therapeutic drugs. The diagnosis of idiopathic sudden deafness can be definitely made when no causes are found. Usually, the impairing mechanism involves the cochlea. The pathophysiology of this sensorineural alteration is still unknown. It is most likely that several mechanisms are associated together, their common point being an impairment to the feedback loop of the organ of Corti. CURRENT KNOWLEDGE AND KEY POINTS: It is very likely that reactivation of neurotropic viruses and/or cochlear ischemia are frequent etiologies. Whatever the cause, the treatment is to be administered urgently, and consists of a high dose corticotherapy at the least. Other treatments have never really proven to be effective. It is secondarily checked that no retrocochlear pathological processes, such as a cerebellopontine angle tumor, is present, in particular in young people. FUTURE PROSPECTS AND PROJECTS: One of the current objectives is to determine when cochlear ischemia is involved, in a mini-invasive manner, such as with laser Doppler flowmetry, so that the treatment can be optimized. From a therapeutic point of view, early acoustic protection has been proven to be effective in cases of cochlear ischemia in small laboratory animals. Its efficacy in case of sudden deafness, non-exclusive of other causes than ischemia, is being assessed in a multicentric project. PMID- 11928378 TI - [Measurement of arterial distensibility by the QKd method a new vascular marker]. AB - INTRODUCTION: Distensibility is the ability of large elastic arteries to increase in diameter from diastole to systole. Pulse wave velocity (PWV) is one of the ways to measure this parameter. Several techniques, including QKd, are able to measure PWV. PURPOSE: QKd is the time interval between the Q wave on EKG and auscultation of the second Korotkoff's sound at the brachial artery. QKd is measured by a specific apparatus that registers ambulatory blood pressure as well as EKG (normal > 200 ms). Arterial distensibility seems to be able to predict cardiovascular morbidity and QKd has been demonstrated to predict such morbidity in a sample of elderly hypertensives. Currently the relationship between QKd and prognosis is under investigation in systemic sclerosis (ERAMS study). CONCLUSION: QKd is a noninvasive ambulatory method that measures arterial distensibility as well as blood pressure. PMID- 11928379 TI - [Anti-TNF alpha monoclonal antibodies (infliximab) and tuberculosis: apropos of 3 cases]. AB - INTRODUCTION: Monoclonal TNF alpha antibodies are a new treatment of severe rheumatoid arthritis. One of the possible side effects is the appearance of opportunistic infections. We report here on three cases of disseminated tuberculosis observed in patients undergoing treatment with infliximab. EXEGESIS: A 45-year-old woman, treated with infliximab, was hospitalised after five infusions for fever and dyspnoea. The exams showed pulmonary and peritoneal tuberculosis. The second case is a 75-year-old woman whose symptoms were fever, cough and cervical adenopathy after three infliximab infusions. Diagnosis was disseminated tuberculosis. The third case is a 59-year-old man who was hospitalised for an infectious syndrome with dyspnoea, after two infliximab infusions. We discovered pulmonary tuberculosis. CONCLUSION: These three cases added to the 68 cases of tuberculosis registered with the treatment of infliximab. This confirms the risk of severe opportunist infectious side effects. TNF alpha is a cytokine which has anti-infectious properties. These tuberculoses are severe and generalized. It is recommended to search for an active or latent tuberculosis before beginning treatment with infliximab, and to check these patients frequently. PMID- 11928380 TI - [Tracheobronchial amyloidosis: apropos of 2 cases]. AB - INTRODUCTION: Tracheo-bronchial amyloidosis is an uncommon localized form of amyloidosis. We report two new cases. EXEGESIS: Two patients had developed expiratory dyspnea for several months. CT-scan and flexible bronchoscopy confirmed tracheal narrowing and a diagnosis of tracheo-bronchial amyloidosis was made by tissue biopsies. The immunohistochemical type was AL in one case, undetermined in the other case. There was no argument for systemic involvement. The two patients benefited from bronchoscopic dilatation. This treatment improved clinical symptoms and pulmonary function tests with a follow up of 12 and 18 months respectively. CONCLUSION: Tracheo-bronchial amyloidosis is a localised form of amyloidosis with various respiratory symptoms. Diagnosis is made by CT scan and flexible bronchoscopy that allows biopsies. Immunohistochemical type is more often AL. Recurrence, respiratory insufficiency and tracheo-bronchial metaplasia are the most important complications. Treatment consists of bronchoscopic dilatation or excision, and bronchoscopic laser-YAG. Pulmonary function testing allows precise follow-up. PMID- 11928381 TI - [Combined sclerosis of the spinal cord revealing vitamin B 12 deficiency: geriatric characteristics apropos of a case evaluated by MRI]. AB - INTRODUCTION: Cobalamin deficiency is common in the elderly. However, most of the patients are asymptomatic or present with few symptoms. A subacute combined degeneration of the spinal cord accounts for only 10% of the neurological complications. Revealing forms of this myelopathy are exceptional, and were rarely documented by magnetic resonance imaging (MRI) study. EXEGESIS: We report a case of subacute combined degeneration of spinal cord in an 81-year-old woman evaluated by a spinal cord MRI. We discuss the role of the spinal cord radiographic study in the diagnosis, the etiopathogeny of cobalamin deficiency and the benefit of vitamin supplementation even at a late stage of the disease in geriatrics patients. CONCLUSION: The spinal cord MRI is useful in the diagnosis of late-form combined spinal cord degeneration. The high frequency of atrophic gastritis in elderly prevents the identification of a deficiency of the cobalamin etiology. The treatment relies on vitamin therapy even in the late stages. The severe disability of cobalamin deficiency neurological complications must encourage an earlier diagnosis in elderly patients. PMID- 11928383 TI - [Multinodular hepatopathy]. PMID- 11928382 TI - [Candidemia in elderly people: apropos of 4 cases]. AB - INTRODUCTION: Candidemia, principally affecting neutropenic patients in departments of oncohematology and frail patients in intensive care units, can also be observed in frail elderly people in geriatrics. EXEGESIS: Authors report four observations of candidemia diagnosed in elderly dependent patients having several different diseases. Clinical sign was a persistent or recurrent fever after a wide-spectrum antibiotic therapy. Patients were treated by fluconazole leading to negative blood cultures in several days. Three out of four patients died within the weeks following antifungal therapy due to severity of associated diseases. CONCLUSION: These observations show that a diagnosis of candidemia should be made when a persistent fever is observed in a frail elderly person. Fluconazole, as efficient as amphotericin B and well tolerated by elderly people, should be the first treatment of candidemia in non-neutropenic patients. PMID- 11928384 TI - [Pulmonary arterial aneurysms revealing Behcet's disease: from diagnosis to treatment]. PMID- 11928385 TI - [Association of seborrheic pemphigus and Hashimoto autoimmune thyroiditis. Apropos of a further case]. PMID- 11928386 TI - [Tumor lysis syndrome with symptomatic hyperuricemia after a first chemotherapeutic treatment of undifferentiated carcinoma of the cavum]. PMID- 11928389 TI - [From inquiry to interactive communication]. AB - Three functions of communication are distinguishable in the practice of dentists: the function directly related to the treatment, the function related to 'informed consent' and the preventive function. For these three functions different communication-models are applicable: communication as transfer-of-information, communication as persuasion and communication as interaction. It is argued that the interaction model of communication is the most promising model nowadays. PMID- 11928388 TI - [Masticatory muscles. Part VIII. Reflexes in the masticatory system]. AB - A reflex is an involuntary response on a specific nerve stimulus. Several types of receptors are present in the masticatory system which can be involved in reflex activity. A short mechanical tap on an incisor, for example, evokes a specific pattern of several so-called exteroceptive reflexes in the electromyographic (EMG) activity of jaw-closing muscles, which includes periods with decreased activity (inhibitory reflex) as well as increased activity (excitatory reflex). In this case, periodontal mechanoreceptors are detecting the stimulus. The second type of reflex that is dealt with is the jaw-jerk reflex as an example of a proprioceptive reflex. A transient increase in length of a jaw closing muscle, detected by length sensors (muscle spindles), evokes an excitatory reflex in the EMG. Dependent on the motor task, the amplitude of both reflexes is modified. The various reflex mechanisms play an important role in normal physiological functions such as posture control and chewing. PMID- 11928390 TI - [The dentist as communicator: the law]. AB - Communication with the patient in dentistry: what the law requires. What does the law require with respect to the communication of dentists with their patients? The answer is first of all to be found in the Act on the medical contract, which is incorporated in the Dutch Civil Code, and applies also to dentistry. In this article, that act is discussed with a view to dental practice. Particular attention is given to the content and extent of the duty to provide information, and to the liability of the dentist who acts negligently on this point. PMID- 11928387 TI - [Acquired angioneurotic edema. Apropos of a case]. PMID- 11928391 TI - [Dental foci. Role, treatment and prophylaxis in patients at risk]. AB - A dental focus usually is a localized chronic infection that under certain circumstances may result in severe local or systemic disease. The most important dental foci are periodontitis, periapical lesions, advanced carious lesions, nonvital pulp, partially impacted teeth and root tips. Local effects of dental foci particularly are processes that may come to expression because of a compromised immunological defence, such as osteoradionecrosis. Systemic effects are mainly caused by transient bacteraemia which can occur spontaneously out of dental foci or after manipulations such as brushing, flossing and dental treatment. Well known examples are infectious endocarditis, fever during chemotherapy and hematogenous infections of total joint prostheses. For all patients at risk (a.o. endocarditis, endoprosthesis, chemotherapy, radiotherapy) it is important that dental foci are treated. Because in most patients the risk factors are present lifelong, a healthy dentition and a healthy periodontium are the best way of prevention. PMID- 11928392 TI - [Relapse after endodontic treatment: endodontic retreatment or apex resection?]. AB - In many cases, unsuccessful endodontic treatment is followed by apicoectomy. Endodontic retreatment, however, could be an appropriate alternative for apicoectomy more frequently. In this paper indications, specific problems and treatment methods are presented for endodontic retreatment, apicoectomy and intentional replantation. PMID- 11928393 TI - [Acute gingivitis: think about leukemia]. PMID- 11928394 TI - [Malaria]. PMID- 11928395 TI - [Is there a place for computer-assisted Post-Academic Dental Education (PAOT)? A survey from the period 1998-2008]. AB - OBJECTIVE: To estimate the role of computer-CPED in the Netherlands to the year 2008. DESIGN: Computer-assisted learning is currently employed in many modern undergraduate dental curricula. It could also play an important role in providing continuing professional education in dentistry (CPED) to general dental practitioners. Computer-CPED offers teacher-independent and individualized interaction by distance learning. METHODS: A simulation study was conducted. Entry data were either derived from publications or were estimated. RESULTS: The results indicate that the demand for CPED will exceed the supply by 2002. CONCLUSION: The introduction of Computer CPED may substantially contribute to the supply of CPED-courses in de first decade of the next century. By 2008 about 25% of the CPED-courses could be provided by computer education. PMID- 11928396 TI - [Guided bone regeneration for dental implantology]. AB - Guided bone regeneration nowadays offers good possibilities to repair local bone defects. Membranes are used to cover the defects. There are many types available. The anatomy of the defect determines mainly which membrane type should be used or which surgical steps should be followed. Most difficult are the one-wall defects, less difficult are four-wall defects. Guided bone regeneration is predictable, but the procedure is technical sensitive and complications and contaminations will easy occur. All steps need to be done carefully, including patient selection and postoperative follow-up care. Implant insertion into the defect can be done simultaneously but will give a higher risk for complications. For this reason, care should be taken for regeneration procedures in aesthetic demanding areas. PMID- 11928397 TI - [The fate of retained mandibular wisdom teeth]. AB - OBJECTIVE: To describe the fate of retained lower wisdom teeth. DESIGN: An inquiry by telephone was done in patients who were registered five years previously and who at that time had at least one lower wisdom tooth. Questions were asked regarding symptoms and treatment of these teeth. SETTING: Tertiary referral centre. METHODS: Retrospective, descriptive. RESULTS: Fifteen out of 55 teeth were removed within the period of five years. Almost all teeth (93.3%) were removed as a preventive treatment. CONCLUSION: Lower wisdom teeth that have been asymptomatic for a longer period of time have a small chance of becoming symptomatic; removal is not indicated in such circumstances. PMID- 11928398 TI - [Quality monitoring of retraining and continuing education in dentistry]. PMID- 11928399 TI - [An ameloblastoma]. PMID- 11928401 TI - [Masticatory muscles. Part VII. Masticatory muscles and mastication. How do we get small pieces of food?]. AB - One of the functions of the masticatory system is to prepare food for swallowing by crushing it into small pieces to be moistened with saliva. The degree of fragmentation of the food particles depends on factors like the chewing force generated by the closer muscles, the jaw movement, and the morphological aspects of the teeth. The jaw movement is the result of a precise neuromuscular control of the various chewing muscles. Experimental research showed that the muscle activity needed to crush the food particles exists of two components: an anticipating component and a component evoked by the food resistance. The anticipating muscle activity is observed only if food resistance is expected. The muscle activity evoked by the food starts on average 25 ms after food contact. The amount of peripherally induced muscle activity linearly increases as a function of the food resistance. This part of the muscle activity is controlled by sensory information of the masticatory system. PMID- 11928400 TI - [Dutch public health in the future. II]. PMID- 11928402 TI - [The Law of Agreement to Medical Treatment (WGBO) in the dental practice: a survey]. AB - On the basis of some pilot studies and several decisions on cases in the Dutch dental disciplinary jurisdiction issues are discussed about the introduction of new legislation in the field of patients rights in dental practice. The following matters are discussed: complaints of patients about the information they got from dentists; opinions of dentists about issues in the field of informed consent: the obligations of a good dentist; issues concerning the answering of difficult patient questions and problems concerning files of dental patients. It is concluded that there might be a discrepancy between the basic assumptions of the new legislation and the practice of dentistry. PMID- 11928403 TI - [Multiple cysts in the lower jaw]. PMID- 11928404 TI - [Diseases of the pharynx: epiglottitis]. PMID- 11928405 TI - [Masticatory muscles. Part IX. Pain in the jaw muscles]. AB - Masticatory muscle pain is considered as a local expression of myofascial pain. The relationship with Myofascial Pain Syndrome and Fibromyalgia is not well understood. Muscle pain is generated through nociceptors served by small-diameter fibres, where processes of sensitization and neurogenic inflammation are important. In contrast to the 'vicious circle' concept, limitation of movement and loss of muscular power seem to be the result of the pain (pain-adaptation model). The diagnosis of muscle pain is made by algometry, while treatment should be simple, reversible and non-invasive. PMID- 11928406 TI - [Re-restoration decisions]. AB - OBJECTIVE: In this study an attempt was made to explain some of the interobserver variation in re-restoration decision making using. METHOD: A simplified re restoration case was shown to 22 dentists who were asked what treatment decision they would make, and what factors would play a role in their decision. The treatment decisions were compared to the outcomes of a knowledge-system on re restoration decision making. RESULTS: The dentists unanimously selected re restoration as the treatment alternative. Half of the dentists preferred amalgam, and the other half chose composite as restorative material. This variation could be explained almost fully by the dentists' weighs on the factors 'tissue loss during preparation', 'restoration and preservation of the chewing function' and 'prognosis and durability of the restoration'. The knowledge system indicated that more than 80% of the dentists actually should have selected composite resin for a restorative material. CONCLUSIONS: It was concluded that the availability of knowledge and its application can explain variation in re-restoration decisions. Knowledge systems can be used in designing practice guidelines. PMID- 11928407 TI - [Repair of the occlusal plane in a mutilated dentition]. AB - In mutilated dentitions the remaining teeth are often migrated, rotated, or erupted. In such situations it is necessary to restore the plane of occlusion adequately. In this article a method is described to restore this plane with the help of the Camper's plane. An intermediary plane is used, drawn on the study model of maxillary teeth, as well as the 'Correct bite' system. PMID- 11928408 TI - [Periodontal aftercare. Prudence after after traditional surgical therapy and after guided tissue regeneration]. AB - The system of maintenance determines the long term success of periodontal therapy. Guided tissue regeneration enables us to regenerate lost periodontal supporting tissues. Likewise a good maintenance system is very important. Since regeneration of periodontal tissue is a relatively slow process, the direct post operative care differs somewhat from the traditional treatment. Prevention of infection and trauma is of the utmost importance. PMID- 11928409 TI - [Single or double?]. PMID- 11928410 TI - [How do screws work in oral implants (when they are detached)?]. AB - Screw-joint connections are commonly used in oral implantology as a means of connecting various components of an implant system. This article discusses some theoretical and clinical principles with respect to the efficacy of screw joint connections in general and of dental implants in particular. PMID- 11928411 TI - [Long term results of overdentures supported by one-stage ITI-implants]. AB - OBJECTIVE: To evaluate the long-term outcome of overdenture treatment in edentulous patients who received dental implants and an overdenture in the mandible. DESIGN: The patients, endentulous in upper and lower jaw, were treated with 218 one face ITI implants, inserted in the interforaminal part of the mandible and evaluated during the first 80 months after insertion of the implants. SETTING: The patients were asked to fill out a questionnaire concerning denture satisfaction and social comfort. The medical files were used for evaluating the aftercare. RESULTS: 7 implants were lost during the period of 80 months. A minimum of surgical and prosthetic aftercare was necessary to maintain the implants and overdentures. The patient satisfaction of the dentures was high, as well as the social comfort, and there was no significant difference between the results after 19 and 80 months. Complaints about the upper dentures were hardly present. CONCLUSION: Implants inserted in the interforaminal part of the mandible are successful. Oral and social comfort is high and less aftercare is necessary, when a strict recall protocol is taken into account. PMID- 11928412 TI - [Distraction osteogenesis. A new development in craniofacial surgery]. AB - By means of distraction osteogenesis (callus distraction) it is possible to reshape bone by creating stress across an iatrogenic fracture line. Subsequent to the successful use of this method in orthopaedic surgery, the development of small intra-oral distractors has opened up the possibilities for its application in maxillo-facial surgery, and the correction of orthodontic and dental problems. Following corticotomy and the fixing of the distractor, it is usually possible to begin adjustment on the fifth post-operative day. Adjustment of the distractor twice a day results in a daily distraction of 1 mm. Both congenital and acquired deformities can be corrected in this way. Class II-1 malocclusion can be corrected by simultaneous bilateral distraction. It is concluded that distraction would seem to have a place in maxillo-facial surgery. PMID- 11928413 TI - [Acute situations in the dental chair: patient who does not respond well]. AB - In daily practice a dentist may encounter a patient with a diminution of consciousness or being short of breath. In this paper a treatment strategy has been described for diagnosis and treatment of acute situations in daily dental practice. PMID- 11928414 TI - [Coagulation disorders: risk factors for bleeding at dental procedures]. AB - Hereditary and acquired coagulation disorders, following the use of cumarins, increase the risk of bleeding associated with dental procedures. To estimate the risk of bleeding after a dental procedure, it is worthwhile to establish a possible bleeding tendency and the use of medicaments, that affect coagulation. In this article normal hemostasis and coagulation are discussed, followed by hereditary and acquired coagulation disorders. The recommended measures, preceding dental procedures are shown, according to the different disorders and the procedure intended. PMID- 11928415 TI - [Initiation of education reforms]. PMID- 11928416 TI - [An unusual swelling on the tongue]. PMID- 11928417 TI - [HIV-infection and AIDS; an update. I]. PMID- 11928418 TI - [Medical accidents in the dental practice. Survey of 471 dentists in the Netherlands]. AB - OBJECTIVE: To determine frequency and nature of medical accidents in Dutch dental practice in relation to type and time of treatment, with and without the use of the MRRH; frequency and nature of the professional assistance. METHOD: Dentists MRRH-users (n = 51) and control dentists (n = 420) recorded medical accidents by name, using a registration form, followed by an anonymous survey. RESULTS: 91 accidents were reported by name by 471 dentists. This contrasted with 300 accidents recorded by 380 dentists in an anonymous survey. No life threatening accidents were reported. Syncope and hyperventilation were frequent. Most of the accidents occurred during local anaesthesia or during treatment, as the procedure became more stressful. Two-third of the accidents could possibly have been prevented by means of a medical history. Medical assistance was requested in 6% of the cases. CONCLUSION: Life threatening disorders were not reported, possible because in the Netherlands no intravenous sedation or general anaesthetic is used in general dental practice. PMID- 11928419 TI - [Reliability of some clinical parameters in implant dentistry]. AB - OBJECTIVE: This study evaluates if clinical tests that are commonly used to evaluate tissues surrounding natural teeth are equally valid in clinical evaluation of dental implants. DESIGN: A retrospective clinical study. SETTING: Department of Oral and Maxillofacial Surgery, Prosthodontics and Special Dental Care of the University of Utrecht. METHOD: The use of a plaque index, a gingival index, probing depth measurements and the Periotest was evaluated by relating the findings to the image on a radiograph in 16 patients, involving 32 IMZ implants. RESULTS: The four clinical tests show poor sensitivity and in general only fair specificity. CONCLUSION: The parameters are unreliable and unfit for clinical evaluation in implant dentistry during regular recall visits. Radiographs are needed to evaluate the condition of a dental implant and to detect critical marginal bone changes surrounding dental implants at an early stage. PMID- 11928420 TI - [Complaints and complications associated with removal of the mandibular third molar. A prospective clinical study]. AB - OBJECTIVE: To study the incidence of complaints and complications after removal of the mandibular third molar and to evaluate the influence of this procedure on functioning of the patient during the first post-surgical week. DESIGN: Prospective, clinical. SETTING: Department of Oral and Maxillofacial Surgery, University Hospital Groningen. METHODS: Patients referred for removal of a mandibular third molar were asked to return one week after the procedure and to keep a daily record of the use of pain medication, duration of the pain and intensity of the pain. RESULTS: Removal of mandibular third molars resulted in an overall complication rate of 12%. Pain medication was used more frequently and for a longer period by patients with post-surgical complications. Due to complaints following the removal of the mandibular third molar, the mean absence from work was one and a half day. Work was generally resumed with decreased perceived efficiency. CONCLUSION: After this commonly performed procedure in dento-alveolar surgery most of the patients were hampered as a result of pain during the first four post-surgical days. Over 10% of the patients developed complications leading to more frequently and prolonged use of pain medication. Removal of the mandibular third molar gave rise to complaints which influenced the patients relatively strong in their daily functioning. PMID- 11928423 TI - [Tooth of time]. PMID- 11928422 TI - [Adjustment to fluoride advice]. PMID- 11928421 TI - [Overdenture prostheses on implants versus complete dentures]. AB - OBJECTIVE: To establish the treatment outcome of implant retained overdentures (IRD) versus complete dentures (CD) in which the outcome assessment focuses on the patient's subjective evaluation ('denture-satisfaction and chewing ability'). DESIGN: A multicenter randomized clinical trial (prospective). SETTING: The department of Oral Function and Prosthetic Dentistry of the University of Nijmegen, the department of Oral and Maxillofacial Surgery of the University Hospital Nijmegen and the department of Oral and Maxillofacial Surgery and Maxillofacial Prosthodontics of the University Hospital Groningen. METHODS: In this study 151 patients with severely resorbed mandibles participated, treatment had been assigned according to a balanced allocation method. Denture satisfaction and chewing ability were assessed using questionnaires before and 1 year after treatment. RESULTS: Before treatment all patients were dissatisfied with their mandibular denture and they could hardly chew tough or had food. One year after treatment the IRD-group was satisfied with their mandibular denture, whereas only one third of the CD-group was satisfied. With respect to the chewing ability the IRO-group scored significantly better than the CD-group (p < or = 0.0001). CONCLUSION: For patients with a severely resorbed mandible, overdentures retained by dental implants appear to provide a more satisfactory solution to their denture-related problems. PMID- 11928424 TI - [Piercing as automutilation?]. PMID- 11928425 TI - [HIV-infection and AIDS; an update. II]. PMID- 11928426 TI - [Templates as an aid in implantology]. AB - The number, position and angulation of implants are of major importance in loading of implants and surrounding bone and in esthetics. Especially in single tooth and partially edentulous cases correct positioning of implants takes a great deal of care. A template is a device that helps to achieve the proper amount of bone and the correct position and angulation of implants. The design and fabrication of radiographic templates, diagnostic templates and surgical templates are described, both in general and specifically for overdentures and non-removable implant-supported restorations. PMID- 11928427 TI - [Reconstruction of the severely resorbed mandible using autologous bone and implants. An evaluation after 5 years]. AB - OBJECTIVE: Evaluation of edentulous patients with severely resorbed mandibles simultaneously treated with bone out of the iliac crest and implants. DESIGN: Prospective. MATERIAL: Thirty-one females between 42 and 61 years of age, were surgically treated and provided with a bar construction or attachments with an overdenture. METHODS: Orthopantomograms were made after 3, 6, 12, 24 and 60 months. Bone resorption was measured mesial and distal from each implant. RESULTS: Eight implants out of 78 inserted implants were lost. The average bone resorption was 0.6 mm after 3 months and 4.4 mm after 60 months. In some patients no bone resorption was found; in other patients the augmented bone was completely lost. CONCLUSION: The described surgical treatment did not give reliable results. PMID- 11928428 TI - [Syndromes 2. Pfeiffer syndrome]. AB - Acrocephalosyndactylias are syndromes characterized by abnormalities of the head (craniosynostosis), the face (hypertelorism, retromaxillism), hands and feet (cutaneous or bony syndactyly). Inheritance is autosomal dominant, but spontaneous cases are described also. The group is divided into several syndromes with varying penetrance and expressivity. As an example of an acrocephalosyndactylia is the Pfeiffer syndrome presented. PMID- 11928429 TI - [Dentists' opinion about their dental education in ACTA from 1990 to 1997]. AB - OBJECTIVE: To assess the dentists' opinion on their (under)graduate dental education. DESIGN: Descriptive survey, questionnaire. SETTING: Academic Centre for Dentistry Amsterdam, The Netherlands. METHODS: 432 dentists who were graduated from the Academic Centre for Dentistry Amsterdam between 1989-1997, were asked to give their opinion on their dental education. RESULTS: From all graduates 43.1% thought the treatment of patients as practiced during their training didn't correspond with the treatment of patients in their present practice. However 75.9% of the respondents thought they had gained enough experience during their dental education to enable them to work as a general practitioner. CONCLUSION: The results show that most respondents were content with the received dental education. The results correspond largely with the results found in a similar survey done in 1990. PMID- 11928430 TI - [Complications after removal of third molar in the lower jaw]. PMID- 11928431 TI - [Tuberculosis. 2]. PMID- 11928432 TI - [Anxious children. Evaluation after treatment at a special dental care center]. AB - OBJECTIVE: Evaluation of dental behaviour of anxious children after treatment at a Special Dental Care Centre. DESIGN: Retrospective, descriptive study. SETTING: Dutch Special Dental Care Centre (SBT). METHODS: A representative sample of parents was interviewed 1-2 years after their child's treatment at the centre about their present behaviour with respect to the home dentist. RESULTS: Results were analyzed using Analysis of Variance. Results indicate that within 1-2 years more than 90% of the children visited a home dentist, 80% of them had to be treated curatively and 60% of the treatment was performed using local anaesthesia. Retrospectively, the parents report a decrease of their child's dental anxiety, when leaving the SBT. The level of anxiety is unchanged after 1-2 years visiting a home dentist. For the children there was a significant relation between dental anxiety as reported by the parents and not visiting a home dentist. The high prevalence of curative treatment might indicate that children in this group belong to a caries risk group and might need extra preventive attention. CONCLUSIONS: After the initial treatment at the special dental care centre most children return to their home dentist. The children in this group referred for dental anxiety, belong to a caries risk group and might need extra preventive attention. PMID- 11928434 TI - [Syndromes 3. Velo-cardio-facial (VCF/Shprintzen) syndrome]. AB - The clinical symptomatology of the velo-cardio-facial (VCF/Shprintzen) syndrome is briefly described. In VCF syndrome a microdeletion of chromosome 22q11.2 can be found, as well as in a number of other clinical entities, leading to the conclusion that it is probably better to speak of the 22q11.2 clinical spectrum. PMID- 11928433 TI - [Dental caries, dental fluorosis and use of fluoride tablets. Results of survey by 8, 14, and 20 year old insurance patients]. AB - OBJECTIVE: To assess the prevalence of fluorosis at the age of 8, 14 and 20 years; to evaluate the prevalence of caries and fluorosis at the age of 8 and 14 years in relation tot the use of fluoride tablets during the first 5 years of age. DESIGN: Retrospective study. SETTING: The investigation was carried out in four cities in the Netherlands. METHODS: Caries and fluorosis data were obtained in 8-, 14- and 20-year-old persons by clinical examination. For 8- and 14-year olds results were compared with data about the use of fluoride tablets during the first 5 years of life. These data were obtained by questionnaires completed by the parents when the child was 5 years old. RESULTS: Prevalence of fluorosis in permanent teeth of 8-, 14 and 20-year-olds was 16%, 31% and 26% respectively. Univariate analysis revealed a statistically significant relation between the frequency of the use of fluoride tablet and caries prevalence in deciduous and permanent teeth on the age of 8. Prevalence of fluorosis and fluoride tablet use did not show such a relation. These findings were confirmed by logistic regression analysis, using frequency of tooth brushing on the age of 5 as possible confounding factor. CONCLUSION: In this group of children the use of fluoride tablets during the first 5 years of life does not seem to increase the prevalence of fluorosis. PMID- 11928435 TI - [Spring congress NVT: 'Special dentistry: if possible a general practitioner, if needed an institution]. PMID- 11928436 TI - [Mouth dryness due to unknown cause?]. PMID- 11928437 TI - [Polio]. PMID- 11928438 TI - [Halitosis, foetor ex ore]. AB - OBJECTIVE: To review the results of a multidisciplinary approach on halitosis. DESIGN: Descriptive. SETTING: University Hospital Leuven, Belgium. METHODS: Review of the data on etiology, diagnosis and therapy of halitosis, obtained by a multidisciplinary team consisting of ENT specialists, periodontologists and a psychiatrist. Malodour was confirmed by a halimeter. RESULTS: 491 patients, equally males and females, with a concentration between 20-50 years of age were seen. Oral causes (87%) were due to tongue coating (51%), gingivitis (17%), periodontitis (15%) or combinations 17%). The other 13% causes were due to ENT related problems (4%), both ENT and oral (3%), digestive tract (1%) and presumed psychic (5%). CONCLUSION: Halitosis is a rather unknown medical problem that often can be solved by a multidisciplinary approach. PMID- 11928439 TI - [Assessment of caries activity and caries risk]. AB - This paper reviews the present knowledge regarding methods of caries prediction. The clinician's ability to predict caries by examination and on the basis of knowledge of patient's attitude and background functions relatively well. Bacterial and salivary tests do not contribute significantly to this. The caries present at the moment of examination, the cariesexperience, is the best predictor of caries among children. PMID- 11928440 TI - [A numb chin and bottom lip: a closer look at the symptoms]. AB - The numb chin syndrome, not due to trauma, dental pathology, preprosthetic surgery or orthognatic surgery, is an uncommon pathological condition. In patients with a numb chin a malignant disease or a metastasis of an (un)known malignancy has to be considered. PMID- 11928441 TI - [Syndromes 4. Sanfilippo syndrome]. AB - The Sanfilippo syndrome belongs to the mucopolysaccharidoses. In this syndrome heparansulfate accumulates in the lysosomes because of an enzymatic disorder in the degradation of mucopolysaccharides. The result is a function disorder of the involved tissues. The Sanfilippo syndrome is characterized by a combination of mental deterioration from the third year of life, hepatosplenomegaly and a typical facial appearance. These characteristics and the implications for dental treatment are discussed. PMID- 11928442 TI - [Dentes natales]. AB - As an introduction, this article describes the circumstances under which dental anatomy was taught to first year students at Utrecht University on November 26, 1945, as this led to the observation of a dens natalis. Relevant literature is discussed concerning the significance of the dens natalis for the child's future and for the family. Investigations into the frequency of the phenomenon are mentioned. Variations of the phenomenon are discussed in two cases. An overview of feeding and eating difficulties and measures to be taken concludes the article. PMID- 11928443 TI - [An easily bleeding swelling of the gingiva in the upper front]. PMID- 11928444 TI - [Breast cancer]. PMID- 11928445 TI - [Geriatric dentistry. Dental health care for the elderly]. PMID- 11928446 TI - [Aging, a frightening image of dental health?]. AB - The combination of decreasing oral self-care and increasing risks of oral health make elderly people an oral health risk-group. In many elderly people above the age of approximately 75 the decline in functions culminates in infirmity. Frail and infirm elderly people require geriatric care, but often oral health care is not an integral part of geriatric care. Geriatric oral health care could be promoted and encouraged by dental hygienists. Unless geriatric oral health care will be promoted, ageing and especially dependent ageing seems a spectre of oral health. PMID- 11928447 TI - [The tooth of time, aging and health]. AB - Older patients are becoming significantly more important in the health care service. Important topics in geriatric medicine are discussed such as characteristics of geriatric patients, the concept of 'normal' and 'successful' ageing, age discrimination and the multidisciplinary geriatric team. These concepts may also be useful in geriatric dentistry. PMID- 11928448 TI - [The dentist in the circle of care of dependent elderly]. AB - By preventing avoidable causes of death, man usually dies at advanced age after a prolonged period of increasing dependency often in homes for the elderly and in nursing homes. This dependency means problems in self care, walking, cognition, communication and in oral health. Geriatric dentistry can be incorporated in a model arranging care (the care circle) in which the dentist operates independently, or organised in teams with other caregivers or central directed by a nursing home doctor (so-called multidisciplinary dentistry). Multidisciplinary dentistry means that the dentist is a member of a multidisciplinary team, contributing to the reduction of individual problems with eating, drinking and oral health. The dentist involves the team in the indications of dental treatment. He operates within the rules of recent Dutch legislation. These laws are based on self-determination of the patient, institutional quality-assurance and individual skills-guarantees of the care provider. PMID- 11928449 TI - [Oral health care for the elderly. More than care for the mouth only]. AB - Dental treatment is changing to oral health care, integrating curative treatment and care. This development is based on the present state of oral health of elderly people. In the future, elderly persons are expected to receive adequate oral health care. This requires gerodontic knowledge and skills, as most oral health problems of elderly persons are chronical and related to ageing processes. For about 25% of elderly people substantially adapted oral health care is needed. This geriatric oral health care can be best provided in collaboration between dentists, dental hygienists and other careproviders. This procedure is already common practice in the care for the chronically ill. Then oral health care will become more than care for oral health. PMID- 11928450 TI - [Geriatric dentistry in dental education]. AB - Geriatric patients have both special medical problems (problems with aging and general health problems) and specific dental problems. This requires special skills of dentists who are treating these patients. The medical problems require a dentist with a wide medical education and a friendly attitude towards his patients. The dental problems require a dentist who is more problem orientated on his way of treatment. Both aspects are present in the theoretical teaching in most of the dental schools. For treatment of institutionalized geriatric patients it is advocated to train specialists in geriatric dentistry taking into account the need for a multidisciplinary and specific approach. PMID- 11928451 TI - [Treatment of anxious patients in special dental care centers. From local initiatives to organized dentistry]. AB - OBJECTIVE: Since the eighties, dental treatment of extremely anxious adult patients became possible in centres for special dental care. In 1996 regulations were formulated about dental treatment for extremely anxious dental patients in these centres. The aim of the present study is to determine how dental treatment of these patients was organised before the regulations were formulated. DESIGN: Descriptive study. SETTING: Twelve centres for special dental care where the largest numbers of anxious patients were treated. METHODS: Semi-structured interviews were held with dentists working in these centres. RESULTS: The results indicate that not (yet) everywhere dentists work according to the regulations. In addition, questionnaires to determine the dental anxiety level and the indications for the different treatment modalities differed between the centres. CONCLUSION: The majority of the centres do not have the capacity to treat the number of patients seeking treatment. It is recommended that research should determine the efficacy of treatment in the centres and should assess which treatment modality is most appropriate for which patient. PMID- 11928452 TI - [Syndromes 5. Williams-Beuren Syndrome]. AB - A characteristic facial appearance, mental retardation, growth deficiency, cardiovascular anomalies, and infantile hypercalcemia are major features of the Williams-Beuren syndrome. The dentist can contribute to the (early) diagnosis of this disorder. PMID- 11928453 TI - [Ectopic geographic tongue]. PMID- 11928454 TI - [Prostate cancer]. PMID- 11928455 TI - [Women and dentistry. Introduction]. PMID- 11928456 TI - [The ambiguity of the difference]. AB - The article concentrates on the process of diagnosing disease, especially of common health problems. The biomedical concept dominates medicine in our days. However, the majority of the complaints presented to the general practician cannot be classified as distinct disease entities. An integral concept is far more suitable to integrate biomedical and psychosocial factors. In this way biomedical aspects acquire significance within an individual life story and social context in which gender plays a role. That is why the integral concept is more adequate to focus on sex differences in the diagnostic process. The article describes the impediments in this process, mainly due to the fact the prevailing concepts of disease ignore sex differences. PMID- 11928457 TI - [Professional practice by female dentists in the Netherlands]. AB - OBJECTIVE: Obtaining information about differences between male and female dentists in practising dentistry and about changes in practice arrangements. DESIGN: Descriptive analysis, using SPSS (CROSSTABS, ANOVA, HOMALS), based on information on the way(s) dentists practice their profession. SETTING: Department of Research of the Dutch Dental Association (NMT), Nieuwegien, The Netherlands. METHODS: A survey by means of a questionnaire, among a representative sample of 229 female dentists and a representative sample of 483 male dentists. RESULTS: Female dentists have on average fewer patients than male dentists, spend fewer working-hours weekly at their chair and practice more often in some form of partnership. Females have a spouse working in the dental practice less often than males and, while the spouse of males most often works as a dental assistant, the spouse of female dentists is most often also a dentist himself. Compared with male dentists, in 1994 female dentists more often had to arrange cover by a locum while in 1995 they had to take more days of leave due to personal illness or exceptional circumstances. CONCLUSION: In the Netherlands there are substantial differences in the practice of the profession by male and female dentists. This is important for the future policy of the NMT with regard to the required manpower in dentistry. PMID- 11928458 TI - [The competence of a female. Battle against the rise of the female dentist]. AB - The refusal of the Dutch Dental Society in 1898 to admit three female dentists to their ranks, was the reason for a polemic. Two important male dentists both wrote a pamphlet considering the competence of women for the study and the practice of learned professions like dentistry. Moreover, they deliberated the value of study and practice for both women and society. The authors are strongly influenced by Social Darwinism and their arguments would now be described as male chauvinism. They admit that women could do the job as well as men, if only after a long evolution. But that evolution is not to be welcomed at all: the best attribute of woman (maternal love) would be frustrated and her highest task (child care) would not be fulfilled. Her femininity would disappear and the new 'man-woman' would not be desired by men, which would be detrimental to the institution of marriage. Already now, they argue, marriages are diminishing, lose quality and divorces are on the increase. They know the cause of all evil: feminism and the women's rights movements. PMID- 11928459 TI - [Working with mercury: cause of infertility in women?]. AB - According to animal experiments, case reports and epidemiological studies, exposition to a high concentration of metallic mercury vapour may cause an increase in reproductive problems. A report on this matter is summarized here as far as it regards women. It seems warranted to conclude that negative reproductive effects are unlikely unless, due to a failing hygiene, the concentration of mercury vapour exceeds the MAC (maximum approved concentration) for women. However, the relationship between concentrations smaller than females' MAC value and problems with cycle and conception need more research. PMID- 11928460 TI - [Transmission of infectious oral diseases from mother to child]. AB - The relation between oral microbes of mother and child encompasses several aspects. It has been shown that a bad periodontal condition is a risk factor for low birth weight. The bacterium that is most specifically associated with dental caries, Streptococcus mutans, can be transmitted between mother and child; it has been suggested that the age of colonization with this bacterium is a risk factor for caries in the child. Also periodontal bacteria can be transmitted between mother and child. It remains to be elucidated whether this is an additional risk factor for the development of periodontal disease. PMID- 11928461 TI - [Oral health and gender]. AB - Recently performed epidemiological studies in the Netherlands did not show statistical significant differences between boys and girls or men and women with regard to the prevalence of caries, periodontal diseases and toothlessness. In adults it was found that the mean number of untreated caries lesions was smaller in women than in men, possibly as a result of the lower percentage of women not attending a dentist regularly. Furthermore, the prevalence of plaque and calculus was lower among women, probably as a result of better oral hygiene habits. PMID- 11928462 TI - [Dental care frequency of female patients in the Netherlands]. AB - OBJECTIVE: Investigate whether there are differences between men and women in dental attendance and in the consumption of dental care. DESIGN: Descriptive analysis, using SPSS (ANOVA, NPAR TEST), based on information on the treatments that are performed for dentate patients. SETTING: Department of Research (afd. O&I) of the Dutch Dental Association (NMT), Nieuwegein, The Netherlands. METHODS: From the practice-administration of 202 dentists, anonymous information was gathered on the treatments these dentists performed for 7698 dentate patients in the years 1993 to 1996. RESULTS: The results indicate that for young people there is only a difference in relation to fillings: girls receive fewer fillings than boys. For adults, however, more differences are apparent. First of all, women visit the dentist for a routine check-up somewhat more often than men. In addition, more X-rays are taken, more instruction is given on oral hygiene and more crowns are made for women, while they receive fewer fillings. CONCLUSION: Adult women and adult men show differences in dental attendance and consumption. These differences are, albeit partially, explained by the fact that adult women are proportionally more often insured for dental costs via the national health service. PMID- 11928463 TI - [Female hormones and oral health]. AB - A common oral manifestation of elevated levels of the ovarian hormones estrogen and progesterone, as seen in pregnancy or oral contraceptive usage, is an increase in gingival inflammation. Estrogen and progesterone probably induce a physiologic vascular phenomenon with swelling and redness. Furthermore, these hormones alter the microenvironment of the oral bacteria so as to promote their growth and cause shifts in their populations. The resulting gingivitis can be avoided or at least minimized by establishing low plaque levels at the beginning of pregnancy or at the start of oral contraceptive therapy. PMID- 11928464 TI - [Osteoporosis]. AB - Osteoporosis is an increasing major skeletal disease leading to disability. It is important to define this disease precisely, because the start is without any symptom in many cases. Causal factors are reviewed as also recognisable risk factors in early stage. The available recent literature and own studies did not show relationship between tooth loss, bone loss of the mandible, periodontitis and osteoporosis. However, corticosteroids diminish mandibular bone mass. Assessment to bone mineral density is a major diagnostic test for osteoporosis prediction. Recent findings in prevention and treatment of osteoporosis are reviewed extensively. The positive results of oestrogen replacement therapy on the oral bone loss and tooth loss from some large observational American studies are interesting. PMID- 11928465 TI - [Facial growth: men and women differ]. AB - In the craniofacial complex there appears to be a sexual dimorphism: men and women not only differ during the 'active growth' period for the timing, amount and localization of growth, but also in the non-active phase thereafter gender differences in facial morphology remain present. In the active growth phase, girls reach their final facial dimensions earlier than boys and boys have larger final facial dimensions than girls. The specific differences in the lateral view of the face are situated in the eminent supraorbital sinus and nose, a larger occipital condyle and mastoid process. In the frontal view, boys show a larger prominence of the zygomatic bones and the gonial region than girls. As far as the 'non-active growth' changes in adulthood are concerned, the differences between men and women are smaller, but consistent and significant. Women have smaller faces (5-9% difference) but there are now configurational differences that appear in the pattern of growth. While men maintain their vertical growth pattern from puberty, women change from a horizontal to a vertical growth pattern during adulthood. PMID- 11928466 TI - [Condylar resorption after osteotomy of the lower jaw]. PMID- 11928467 TI - On a family-based haplotype pattern mining method for linkage disequilibrium mapping. AB - Linkage disequilibrium mapping is an important tool in disease gene mapping. Recently, Toivonen et al. [1] introduced a haplotype mining (HPM) method that is applicable to data consisting of unrelated high-risk and normal haplotypes. The HPM method orders haplotypes by their strength of association with trait values, and uses all haplotypes exceeding a given threshold of strength of association to predict the gene location. In this study, we extend the HPM method to pedigree data by measuring the strength of association between a haplotype and quantitative traits of interest using the Quantitative Pedigree Disequilibrium Test proposed by Zhang et al. [2]. This family-based HPM (F-HPM) method can incorporate haplotype information across a set of markers and allow both missing marker data and ambiguous haplotype information. We use a simulation procedure to evaluate the statistical significance of the patterns identified from the F-HPM method. When the F-HPM method is applied to analyze the sequence data from the seven candidate genes in the simulated data sets in the 12th Genetic Analysis Workshop, the association between genes and traits can be detected with high power, and the estimated locations of the trait loci are close to the true sites. PMID- 11928468 TI - Scoring pairwise genomic sequence alignments. AB - The parameters by which alignments are scored can strongly affect sensitivity and specificity of alignment procedures. While appropriate parameter choices are well understood for protein alignments, much less is known for genomic DNA sequences. We describe a straightforward approach to scoring nucleotide substitutions in genomic sequence alignments, especially human-mouse comparisons. Scores are obtained from relative frequencies of aligned nucleotides observed in alignments of non-coding, non-repetitive genomic regions, and can be theoretically motivated through substitution models. Additional accuracy can be attained by down weighting alignments characterized by low compositional complexity. We also describe an evaluation protocol that is relevant when alignments are intended to identify all and only the orthologous positions. One particular scoring matrix, called HOXD70, has proven to be generally effective for human-mouse comparisons, and has been used by the PipMaker server since July, 2000. We discuss but leave open the problem of effectively scoring regions of strongly biased nucleotide composition, such as low G + C content. PMID- 11928470 TI - Constructing comparative genome maps with unresolved marker order. AB - Comparative genome maps are a powerful tool for interpreting the genomes of related organisms. The species maps which are the input to the process of constructing comparative maps are often themselves constructed from incomplete or inconsistent data, resulting in markers (or genes) whose order is not fully resolved. This incomplete marker order information is often handled by placing markers whose relative order cannot be reliably inferred together in a bin which is mapped to a common location. Previous automated and manual methods have handled such markers in an ad hoc or arbitrary way. We present efficient algorithms for comparative map construction that provide a principled method for handling unresolved marker order. The algorithms are based on a technique for efficiently computing a marker order that optimizes a natural parsimony criterion; in this way, they also yield a working hypothesis about the original incomplete data set. PMID- 11928469 TI - Structure-based comparison of four eukaryotic genomes. AB - The field of comparative genomics allows us to elucidate the molecular mechanisms necessary for the machinery of an organism by contrasting its genome against those of other organisms. In this paper, we contrast the genome of homo sapiens against C. Elegans, Drosophila melanogaster, and S. cerevisiae to gain insights on what structural domains are present in each organism. Previous work has assessed this using sequence-based homology recognition systems such as Pfam [1] and Interpro [2]. Here, we pursue a structure-based assessment, analyzing genomes according to domains in the SCOP structural domain dictionary. Compared to other eukaryotic genomes, we observe additional domains in the human genome relating to signal transduction, immune response, transport, and certain enzymes. Compared to the metazoan genomes, the yeast genome shows an absence of domains relating to immune response, cell-cell interactions, and cell signaling. PMID- 11928471 TI - Representation and processing of complex DNA spatial architecture and its annotated genomic content. AB - This paper presents a new general approach for the spatial representation and visualization of DNA molecule and its annotated information. This approach is based on a biological 3D model that predicts the complex spatial trajectory of huge naked DNA. With such modeling, a global vision of the sequence is possible, which is different and complementary to other representations as textual, linguistics or syntactic ones. The DNA is well known as a three-dimensional structure. Whereas, the spatial information plays a great part during its evolution and its interaction with the other biological elements This work will motivate investigations in order to launch new bioinformatics studies for the analysis of the spatial architecture of the genome. Besides, in order to obtain a friendly interactive visualization, a powerful graphic modeling is proposed including DNA complex trajectory management and its annotated-based content structuring. The paper describes spatial architecture modeling, with consideration of both biological and computational constraints. This work is implemented through a powerful graphic software tool, named ADN-Viewer. Several examples of visualization are shown for various organisms and biological elements. PMID- 11928472 TI - Pairwise RNA structure comparison with stochastic context-free grammars. AB - Pairwise stochastic context-free grammars ("Pair SCFGs") are powerful tools for finding conserved RNA structures, but unconstrained alignment to Pair SCFGs is prohibitively expensive. We develop versions of the Pair SCFG dynamic programming algorithms that can be conditioned on precomputed structures, significantly reducing the time complexity of alignment. We have implemented these algorithms for general Pair SCFGs in software that is freely available under the GNU Public License. PMID- 11928473 TI - Estimation of genetic networks and functional structures between genes by using Bayesian networks and nonparametric regression. AB - We propose a new method for constructing genetic network from gene expression data by using Bayesian networks. We use nonparametric regression for capturing nonlinear relationships between genes and derive a new criterion for choosing the network in general situations. In a theoretical sense, our proposed theory and methodology include previous methods based on Bayes approach. We applied the proposed method to the S. cerevisiae cell cycle data and showed the effectiveness of our method by comparing with previous methods. PMID- 11928474 TI - Singular value decomposition regression models for classification of tumors from microarray experiments. AB - An important problem in the analysis of microarray data is correlating the high dimensional measurements with clinical phenotypes. In this paper, we develop predictive models for associating gene expression data from microarray experiments with such outcomes. They are based on the singular value decomposition. We propose new algorithms for performing gene selection and gene clustering based on these predictive models. The estimation procedure using the regression models occurs in two stages. First, the gene expression measurements are transformed using the singular value decomposition. The regression parameters in the model linking the principal components with the clinical responses are then estimated using maximum likelihood. We demonstrate the application of the methodology to data from a breast cancer study. PMID- 11928475 TI - Automatic annotation of genomic regulatory sequences by searching for composite clusters. AB - A new method was developed for revealing of composite clusters of cis-elements in promoters of eukaryotic genes that are functionally related or coexpressed. A software system "ClusterScan" have been created that enables: (i) to train system on representative samples of promoters to reveal cis-elements that tend to cluster, (ii) to train system on a number of samples of functionally related promoters to identify functionally coupled transcription factors; (iii) to provide tools for searching of this clusters in genomic sequences to identify and functionally characterize regulatory regions in genome. A number of training samples of different functional and structural groups of promoters were analysed. Search for composite clusters in human chromosomes 21 and 22 reveals a number of interesting examples. Finally, a decision tree system was constructed to classify promoters of several functionally related gene groups. The decision tree system enables to identify new promoters and computationally predict their possible function. PMID- 11928476 TI - EULER-PCR: finishing experiments for repeat resolution. AB - Genomic sequencing typically generates a large collection of unordered contigs or scaffolds. Contig ordering (also known as gap closure) is a non-trivial algorithmic and experimental problem since even relatively simple-to-assemble bacterial genomes typically result in large set of contigs. Neighboring contigs maybe separated either by gaps in read coverage or by repeats. In the later case we say that the contigs are separated by pseudogaps, and we emphasize the important difference between gap closure and pseudogap closure. The existing gap closure approaches do not distinguish between gaps and pseudogaps and treat them in the same way. We describe a new fast strategy for closing pseudogaps (repeat resolution). Since in highly repetitive genomes, the number of pseudogaps may exceed the number of gaps by an order of magnitude, this approach provides a significant advantage over the existing gap closure methods. PMID- 11928477 TI - The accuracy of fast phylogenetic methods for large datasets. AB - Whole-genome phylogenetic studies require various sources of phylogenetic signals to produce an accurate picture of the evolutionary history of a group of genomes. In particular, sequence-based reconstruction will play an important role, especially in resolving more recent events. But using sequences at the level of whole genomes means working with very large amounts of data--large numbers of sequences--as well as large phylogenetic distances, so that reconstruction methods must be both fast and robust as well as accurate. We study the accuracy, convergence rate, and speed of several fast reconstruction methods: neighbor joining, Weighbor (a weighted version of neighbor-joining), greedy parsimony, and a new phylogenetic reconstruction method based on disk-covering and parsimony search (DCM-NJ + MP). Our study uses extensive simulations based on random birth death trees, with controlled deviations from ultrametricity. We find that Weighbor, thanks to its sophisticated handling of probabilities, outperforms other methods for short sequences, while our new method is the best choice for sequence lengths above 100. For very large sequence lengths, all four methods have similar accuracy, so that the speed of neighbor-joining and greedy parsimony makes them the two methods of choice. PMID- 11928478 TI - Pre-mRNA secondary structure prediction aids splice site prediction. AB - Accurate splice site prediction is a critical component of any computational approach to gene prediction in higher organisms. Existing approaches generally use sequence-based models that capture local dependencies among nucleotides in a small window around the splice site. We present evidence that computationally predicted secondary structure of moderate length pre-mRNA subsequencies contains information that can be exploited to improve acceptor splice site prediction beyond that possible with conventional sequence-based approaches. Both decision tree and support vector machine classifiers, using folding energy and structure metrics characterizing helix formation near the splice site, achieve a 5-10% reduction in error rate with a human data set. Based on our data, we hypothesize that acceptors preferentially exhibit short helices at the splice site. PMID- 11928479 TI - Finding weak motifs in DNA sequences. AB - Recognition of regulatory sites in unaligned DNA sequences is an old and well studied problem in computational molecular biology. Recently, large-scale expression studies and comparative genomics brought this problem into a spotlight by generating a large number of samples with unknown regulatory signals. Here we develop algorithms for recognition of signals in corrupted samples (where only a fraction of sequences contain sites) with biased nucleotide composition. We further benchmark these and other algorithms on several bacterial and archaeal sites in a setting specifically designed to imitate the situations arising in comparative genomics studies. PMID- 11928480 TI - Evidence for sequence-independent evolutionary traces in genomics data. AB - Sequence conservation during evolution is the foundation for the functional classification of the enormous number of new protein sequences being discovered in the current era of genome sequencing. Conventional methods to detect homologous proteins are not always able to distinguish between true homologs and false positive hits in the twilight zone of sequence similarity. Several different approaches have been proposed to improve the sensitivity of these methods. Among the most successful are sequence profiles, multi-linked alignment, and threading. However, evolution might offer up other clues about a protein's ancestry that are sequence independent. Here we report the discovery of two such traces of evolution that could potentially be used to help infer the fold of a protein and hence improve the ability to predict the biochemical function. The first such evolutionary trace is a conservation of fold along the genome, i.e. nearby genes tend to share a fold more often than expected by chance alone--a not unexpected observation, but one which holds true even when no pair of genes being examined share appreciable homology. The second such evolutionary trace is, surprisingly, present in expression data: genes that are correlated in expression are more apt to share a fold than two randomly chosen genes. This result is surprising because correlations in expression have previously only been considered useful for determining biological function (e.g. what pathway a particular gene fits into), yet the observed fold enrichment in the expression data permits us to say something about biochemical function since fold corresponds strongly with biochemical function. Again, the fold enrichment observed in the expression data is apparent even when no pair of genes being examined share appreciable homology. PMID- 11928481 TI - Multiple genome rearrangement by reversals. AB - In this paper, we discuss a multiple genome rearrangement problem: Given a collection of genomes represented by permutations, we generate the collection from some fixed genome, e.g., the identity permutation, in a minimum number of signed reversals. It is NP-hard, so efficient heuristics is important for finding its optimal solution. We at first discuss how to generate two and three genomes from a fixed genome by polynomial algorithms for some special cases. Then based on the polynomial algorithms, we obtain some approximation algorithms for generating two and three genomes in general, respectively. Finally, we apply these approximation algorithms to design a new approximation algorithm for generating more genomes. We also show by some experimental examples that the algorithms are efficient. PMID- 11928482 TI - High speed homology search with FPGAs. AB - We will introduce a way how we can achieve high speed homology search by only adding one off-the-shelf PCI board with one Field Programmable Gate Array (FPGA) to a Pentium based computer system in use. FPGA is a reconfigurable device, and any kind of circuits, such as pattern matching program, can be realized in a moment. The performance is almost proportional to the size of FPGA which is used in the system, and FPGAs are becoming larger and larger following Moore's law. We can easily obtain latest/larger FPGAs in the form off-the-shelf PCI boards with FPGAs, at low costs. The result which we obtained is as follows. The performance is most comparable with small to middle class dedicated hardware systems when we use a board with one of the latest FPGAs and the performance can be furthermore accelerated by using more number of FPGA boards. The time for comparing a query sequence of 2,048 elements with a database sequence of 64 million elements by the Smith-Waterman algorithm is about 34 sec, which is about 330 times faster than a desktop computer with a 1 GHz Pentium III. We can also accelerate the performance of a laptop computer using a PC card with one smaller FPGA. The time for comparing a query sequence (1,024) with the database sequence (64 million) is about 185 sec, which is about 30 times faster than the desktop computer. PMID- 11928483 TI - Glycosylation of proteins: a computer based method for the rapid exploration of conformational space of N-glycans. AB - Inspection of protein databases suggests that as many as 70% of proteins have potential N-glycosylation sites. Unfortunately glycoproteins often refuse to crystallize and NMR techniques do not allow an unambiguous determination of the complete conformation of the sugar part. Therefore, time-consuming complex simulation methods are often used to explore the conformational space of N glycans. The generation of a comprehensive data base describing the conformational space of larger fragments of N-glycans taking into account the effects of branching is presented. High-temperature molecular dynamics simulations of essential N-glycan fragments are performed until conformational equilibrium has been reached. Free energy landscapes are calculated for each glycosidic linkage. All possible conformations for each N-glycan fragment are automatically assigned, ranked according to their relative population and stored in a database. These values are recalled for the generation of a complete set of all possible conformations for a given N-glycan topology. The constructed conformations are ranked according to their energy content. Since this approach allows to explore the complete conformational space of a given N-glycan within a few minutes of CPU-time on a standard PC, it is well suited to be used as a Web Based application. PMID- 11928484 TI - Data standardisation in GlycoSuiteDB. AB - GlycoSuiteDB, a database of glycan structures, has been constructed with an emphasis on quality, consistency and data integrity. Importance has been placed on making the database a reliable and useful resource for all researchers. This database can help researchers to identify what glycan structures are known to be attached to certain glycoproteins, as well as more generally identifying what types of glycan structures are associated with different states, for example, different species, tissues and diseases. To achieve this, a major effort has gone into data standardisation. Many rules and standards have been adopted, especially for representing glycan structure and biological source information. This paper describes some of the challenges faced during the continuous development of GlycoSuiteDB. PMID- 11928485 TI - An automated computer system to support ultra high throughput SNP genotyping. AB - Celera Genomics has constructed an automated computer system to support ultra high-throughput SNP genotyping that satisfies the increasing demand that disease association studies are placing on current genotyping facilities. This system consists of the seamless integration of target SNP selection, automated oligo design, in silico assay quality validation, laboratory management of samples, reagents and plates, automated allele calling, optional manual review of autocalls, regular status reports, and linkage disequilibrium analysis. Celera has proven the system by generating over 2.5 million genotypes from more than 10,000 SNPs, and is approaching the target capacity of over 10,000 genotypes per machine per hour using limited human intervention with state of the art laboratory hardware. PMID- 11928486 TI - Prediction of glycosylation across the human proteome and the correlation to protein function. PMID- 11928487 TI - Mining MEDLINE: abstracts, sentences, or phrases? AB - A growing body of works address automated mining of biochemical knowledge from digital repositories of scientific literature, such as MEDLINE. Some of these works use abstracts as the unit of text from which to extract facts. Others use sentences for this purpose, while still others use phrases. Here we compare abstracts, sentences, and phrases in MEDLINE using the standard information retrieval performance measures of recall, precision, and effectiveness, for the task of mining interactions among biochemical terms based on term co-occurrence. Results show statistically significant differences that can impact the choice of text unit. PMID- 11928488 TI - Creating knowledge repositories from biomedical reports: the MEDSYNDIKATE text mining system. AB - MEDSYNDIKATE is a natural language processor for automatically acquiring knowledge from medical finding reports. The content of these documents is transferred to formal representation structures which constitute a corresponding text knowledge base. The system architecture integrates requirements from the analysis of single sentences, as well as those of referentially linked sentences forming cohesive texts. The strong demands MEDSYNDIKATE poses to the availability of expressive knowledge sources are accounted for by two alternative approaches to (semi)automatic ontology engineering. We also present data for the knowledge extraction performance of MEDSYNDIKATE for three major syntactic patterns in medical documents. PMID- 11928489 TI - Filling preposition-based templates to capture information from medical abstracts. AB - Due to the recent explosion of information in the biomedical field, it is hard for a single researcher to review the complex network involving genes, proteins, and interactions. We are currently building GeneScene, a toolkit that will assist researchers in reviewing existing literature, and report on the first phase in our development effort: extracting the relevant information from medical abstracts. We are developing a medical parser that extracts information, fills basic prepositional-based templates, and combines the templates to capture the underlying sentence logic. We tested our parser on 50 unseen abstracts and found that it extracted 246 templates with a precision of 70%. In comparison with many other techniques, more information was extracted without sacrificing precision. Future improvement in precision will be achieved by correcting three categories of errors. PMID- 11928490 TI - Robust relational parsing over biomedical literature: extracting inhibit relations. AB - We describe the design of a robust parser for identifying and extracting biomolecular relations from the biomedical literature. Separate automata over distinct syntactic domains were developed for extraction of nominal-based relational information versus verbal-based relations. This allowed us to optimize the grammars separately for each module, regardless of any specific relation resulting in significantly better performance. A unique feature of this system is the use of text-based anaphora resolution to enhance the results of argument binding in relational extraction. We demonstrate the performance of our system on inhibition-relations, and present our initial results measured against an annotated text used as a gold standard for evaluation purposes. The results represent a significant improvement over previously published results on extracting such relations from Medline: Precision was 90%, Recall 57%, and Partial Recall 22%. These results demonstrate the effectiveness of a corpus-based linguistic approach to information extraction over Medline. PMID- 11928491 TI - Predicting the sub-cellular location of proteins from text using support vector machines. AB - We present an automatic method to classify the sub-cellular location of proteins based on the text of relevant medline abstracts. For each protein, a vector of terms is generated from medline abstracts in which the protein/gene's name or synonym occurs. A Support Vector Machine (SVM) is used to automatically partition the term space and to thus discriminate the textual features that define sub cellular location. The method is benchmarked on a set of proteins of known sub cellular location from S. cerevisiae. No prior knowledge of the problem domain nor any natural language processing is used at any stage. The method out-performs support vector machines trained on amino acid composition and has comparable performance to rule-based text classifiers. Combining text with protein amino acid composition improves recall for some sub-cellular locations. We discuss the generality of the method and its potential application to a variety of biological classification problems. PMID- 11928492 TI - A thematic analysis of the AIDS literature. AB - Faced with the need for human comprehension of any large collection of objects, a time honored approach has been to cluster the objects into groups of closely related objects. Individual groups are then summarized in some convenient manner to provide a more manageable view of the data. Such methods have been applied to document collections with mixed results. If a hard clustering of the data into mutually exclusive clusters is performed then documents are frequently forced into one cluster when they may contain important information that would also appropriately make them candidates for other clusters. If a soft clustering is used there still remains the problem of how to provide a useful summary of the data in a cluster. Here we introduce a new algorithm to produce a soft clustering of document collections that is based on the concept of a theme. A theme is conceptually a subject area that is discussed by multiple documents in the database. A theme has two potential representations that may be viewed as dual to each other. First it is represented by the set of documents that discuss the subject or theme and second it is also represented by the set of key terms that are typically used to discuss the theme. Our algorithm is an EM algorithm in which the term representation and the document representation are explicit components and each is used to refine the other in an alternating fashion. Upon convergence the term representation provides a natural summary of the document representation (the cluster). We describe how to optimize the themes produced by this process and give the results of applying the method to a database of over fifty thousand PubMed documents dealing with the subject of AIDS. How themes may improve access to a document collection is also discussed. PMID- 11928494 TI - Inferring genotype from clinical phenotype through a knowledge based algorithm. AB - Genomic information is becoming increasingly useful for studying the origins of disease. Recent studies have focused on discovering new genetic loci and the influence of these loci upon disease. However, it is equally desirable to go in the opposite direction--that is, to infer genotype from the clinical phenotype for increased efficiency of treatment. This paper proposes a methodology for such inference. Our method constructs a simple knowledge-based model without the need of a domain expert and is useful in situations that have very little data and/or no training data. The model relates a disease's symptoms to particular clinical states of the disease. Clinical information is processed using the model, where appropriate weighting of the symptoms is learned from observed diagnoses to subsequently identify the state of the disease presented in hospital visits. This approach applies to any simple genetic disorder that has defined clinical phenotypes. We demonstrate the use of our methods by inferring age of onset and DNA mutations for Huntington's disease patients. PMID- 11928493 TI - Pathway logic: symbolic analysis of biological signaling. AB - The genomic sequencing of hundreds of organisms including homo sapiens, and the exponential growth in gene expression and proteomic data for many species has revolutionized research in biology. However, the computational analysis of these burgeoning datasets has been hampered by the sparse successes in combinations of data sources, representations, and algorithms. Here we propose the application of symbolic toolsets from the formal methods community to problems of biological interest, particularly signaling pathways, and more specifically mammalian mitogenic and stress responsive pathways. The results of formal symbolic analysis with extremely efficient representations of biological networks provide insights with potential biological impact. In particular, novel hypotheses may be generated which could lead to wet lab validation of new signaling possibilities. We demonstrate the graphic representation of the results of formal analysis of pathways, including navigational abilities, and describe the logical underpinnings of the approach. In summary, we propose and provide an initial description of an algebra and logic of signaling pathways and biologically plausible abstractions that provide the foundation for the application of high powered tools such as model checkers to problems of biological interest. PMID- 11928495 TI - Towards the prediction of complete protein--protein interaction networks. AB - We present a statistical method for the prediction of protein--protein interactions within an organism. This approach is based on the treatment of proteins as collections of conserved domains, where each domain is responsible for a specific interaction with another domain. By characterizing the frequency with which specific domain--domain interactions occur within known interactions, our model can assign a probability to an arbitrary interaction between any two proteins with defined domains. Domain interaction data is complemented with information on the topology of a network and is incorporated into the model by assigning greater probabilities to networks displaying more biologically realistic topologies. We use Markov chain Monte Carlo techniques for the prediction of posterior probabilities of interaction between a set of proteins; allowing its application to large data sets. In this work we attempt to predict interactions in a set of 40 human proteins, known to form a connected network, and discuss methods for future improvement. PMID- 11928496 TI - Identifying muscle regulatory elements and genes in the nematode Caenorhabditis elegans. AB - We report the identification of several putative muscle-specific regulatory elements, and genes which are expressed preferentially in the muscle of the nematode Caenorhabditis elegans. We used computational pattern finding methods to identify cis-regulatory motifs from promoter regions of a set of genes known to express preferentially in muscle; each motif describes the potential binding sites for an unknown regulatory factor. The significance and specificity of the identified motifs were evaluated using several different control sequence sets. Using the motifs, we searched the entire C. elegans genome for genes whose promoter regions have a high probability of being bound by the putative regulatory factors. Genes that met this criterion and were not included in our initial set were predicted to be good candidates for muscle expression. Some of these candidates are additional, known muscle expressed genes and several others are shown here to be preferentially expressed in muscle cells by using GFP (green fluorescent protein) constructs. The methods described here can be used to predict the spatial expression pattern of many uncharacterized genes. PMID- 11928497 TI - Combining location and expression data for principled discovery of genetic regulatory network models. AB - We develop principled methods for the automatic induction (discovery) of genetic regulatory network models from multiple data sources and data modalities. Models of regulatory networks are represented as Bayesian networks, allowing the models to compactly and robustly capture probabilistic multivariate statistical dependencies between the various cellular factors in these networks. We build on previous Bayesian network validation results by extending the validation framework to the context of model induction, leveraging heuristic simulated annealing search algorithms and posterior model averaging. Using expression data in isolation yields results inconsistent with location data so we incorporate genomic location data to guide the model induction process. We combine these two data modalities by allowing location data to influence the model prior and expression data to influence the model likelihood. We demonstrate the utility of this approach by discovering genetic regulatory models of thirty-three variables involved in S. cerevisiae pheromone response. The models we automatically generate are consistent with the current understanding regarding this regulatory network, but also suggest new directions for future experimental investigation. PMID- 11928498 TI - The ERATO Systems Biology Workbench: enabling interaction and exchange between software tools for computational biology. AB - Researchers in computational biology today make use of a large number of different software packages for modeling, analysis, and data manipulation and visualization. In this paper, we describe the ERATO Systems Biology Workbench (SBW), a software framework that allows these heterogeneous application components--written in diverse programming languages and running on different platforms--to communicate and use each others' data and algorithmic capabilities. Our goal is to create a simple, open-source software infrastructure which is effective, easy to implement and easy to understand. SBW uses a broker-based architecture and enables applications (potentially running on separate, distributed computers) to communicate via a simple network protocol. The interfaces to the system are encapsulated in client-side libraries that we provide for different programming languages. We describe the SBW architecture and the current set of modules, as well as alternative implementation technologies. PMID- 11928499 TI - Genome-wide pathway analysis and visualization using gene expression data. AB - Visualization of results for high performance computing pose special problems due to the complexity and the volume of data these systems manipulate. We present an approach for visualization of c-DNA microarray gene expression data in metabolic and regulatory pathways using multi-resolution animation at different levels of detail. We describe three scoring functions to characterize pathways at the transcriptional level based on gene expression, coregulation and cascade effect. We also assess the significance of each pathway score on the basis of their biological connotation. PMID- 11928500 TI - Exploring gene expression data with class scores. AB - We address a commonly asked question about gene expression data sets: "What functional classes of genes are most interesting in the data?" In the methods we present, expression data is partitioned into classes based on existing annotation schemes. Each class is then given three separately derived "interest" scores. The first score is based on an assessment of the statistical significance of gene expression changes experienced by members of the class, in the context of the experimental design. The second is based on the co-expression of genes in the class. The third score is based on the learnability of the classification. We show that all three methods reveal significant classes in each of three different gene expression data sets. Many classes are identified by one method but not the others, indicating that the methods are complementary. The classes identified are in many cases of clear relevance to the experiment. Our results suggest that these class scoring methods are useful tools for exploring gene expression data. PMID- 11928501 TI - Guiding revision of regulatory models with expression data. AB - BioLingua is a computational system designed to support biologists' efforts to construct models, make predictions, and interpret data. In this paper, we focus on the specific task of revising an initial model of gene regulation based on expression levels from gene microarrays. We describe BioLingua's formalism for representing process models, its method for predicting qualitative correlations from such models, and its use of data to constrain search through the space of revised models. We also report experimental results on revising a model of photosynthetic regulation in Cyanobacteria to better fit expression data for both wild and mutant strains, along with model mutilation studies designed to test our method's robustness. In closing, we discuss related work on representing, discovering, and revising biological models, after which we propose some directions for future research. PMID- 11928502 TI - Discovery of causal relationships in a gene-regulation pathway from a mixture of experimental and observational DNA microarray data. AB - This paper reports the methods and results of a computer-based search for causal relationships in the gene-regulation pathway of galactose metabolism in the yeast Saccharomyces cerevisiae. The search uses recently published data from cDNA microarray experiments. A Bayesian method was applied to learn causal networks from a mixture of observational and experimental gene-expression data. The observational data were gene-expression levels obtained from unmanipulated "wild type" cells. The experimental data were produced by deleting ("knocking out") genes and observing the expression levels of other genes. Causal relations predicted from the analysis on 36 galactose gene pairs are reported and compared with the known galactose pathway. Additional exploratory analyses are also reported. PMID- 11928503 TI - Shallow genomics, phylogenetics, and evolution in the family Drosophilidae. AB - The effects of the genomic revolution are beginning to be felt in all disciplines of the biological sciences. Evolutionary biology in general, and phylogenetic systematics in particular, are being revolutionized by these advances. The advent of rapid nucleotide sequencing techniques have provided phylogenetic biologists with the tools required to quickly and efficiently generate large amounts of character information. We use family Drosophilidae as a model system to study phylogenetics and genome evolution by combining high throughput sequencing methods from the field genomics and standard phylogenetic methodology. This paper presents preliminary results from this work. Separate data partitions, based on either gene function or linkage group, are compared to a combined analysis of all the data to assess support on phylogenetic trees. PMID- 11928504 TI - Fast phylogenetic methods for the analysis of genome rearrangement data: an empirical study. AB - Evolution operates on whole genomes through mutations that change the order and strandedness of genes within the genomes. Thus analyses of gene-order data present new opportunities for discoveries about deep evolutionary events, provided that sufficiently accurate methods can be developed to reconstruct evolutionary trees. In this paper we present two new methods of character coding for parsimony-based analysis of genomic rearrangements: one called MPBE-2, and a new parsimony-based method which we call MPME (based on an encoding of Bryant), both variants of the MPBE method. We then conduct computer simulations to compare this class of methods to distance-based methods (NJ under various distance measures). Our empirical results show that two of our new methods return highly accurate estimates of the true tree, outperforming the other methods significantly, especially when close to saturation. PMID- 11928505 TI - A cellular automata approach to detecting interactions among single-nucleotide polymorphisms in complex multifactorial diseases. AB - The identification and characterization of susceptibility genes for common complex multifactorial human diseases remains a statistical and computational challenge. Parametric statistical methods such as logistic regression are limited in their ability to identify genes whose effects are dependent solely or partially on interactions with other genes and environmental exposures. We introduce cellular automata (CA) as a novel computational approach for identifying combinations of single-nucleotide polymorphisms (SNPs) associated with clinical endpoints. This alternative approach is nonparametric (i.e. no hypothesis about the value of a statistical parameter is made), is model-free (i.e. assumes no particular inheritance model), and is directly applicable to case-control and discordant sib-pair study designs. We demonstrate using simulated data that the approach has good power for identifying high-order nonlinear interactions (i.e. epistasis) among four SNPs in the absence of independent main effects. PMID- 11928506 TI - Vertebrate phylogenomics: reconciled trees and gene duplications. AB - Ancient gene duplication events have left many traces in vertebrate genomes. Reconciled trees represent the differences between gene family trees and the species phylogeny those genes are sampled from, allowing us to both infer gene duplication events and estimate a species phylogeny from a sample of gene families. We show that analysis of 118 gene families yields a phylogeny of vertebrates largely in agreement with other data. We formulate the problem of locating episodes of gene duplication as a set cover problem: given a species tree in which each node has a set of gene duplications associated with it, the smallest set of species nodes whose union includes all gene duplications specifies the locations of gene duplication episodes. By generating a unique mapping from this cover set we can determine the minimal number of such episodes at each location. When applied to our data, this method reveals a complex history of gene duplications in vertebrate evolution that does not conform to the "2R" hypothesis. PMID- 11928507 TI - Screened charge electrostatic model in protein-protein docking simulations. AB - A new method for considering solvation when calculating electrostatics for protein docking is proposed. The solvent-exposed charges are attenuated by induced solvent polarization charges. Modified charges are pre-calculated and the correction doesn't affect the speed of the actual simulation. The new Screened Charge Electrostatic Model (SChEM) results in an improved discrimination of near native solutions from false positives in docking simulations as compared to conventional 'non-solvated' charge assignment. A series of protein-protein complexes were analyzed by running automated rigid-body Monte-Carlo docking simulations using the 3-D coordinates of the unbound components. In all but one case, the use of solvation screened charges for electrostatic calculations helped to improve the rank of the near-native solution after rigid-body simulations. The SChEM also drastically improved the results of the subsequent refinement of the interface side-chains. In all cases the final lowest energy solution was found within 3.0 A r.m.s.d. of the crystal structure. PMID- 11928508 TI - The spectrum kernel: a string kernel for SVM protein classification. AB - We introduce a new sequence-similarity kernel, the spectrum kernel, for use with support vector machines (SVMs) in a discriminative approach to the protein classification problem. Our kernel is conceptually simple and efficient to compute and, in experiments on the SCOP database, performs well in comparison with state-of-the-art methods for homology detection. Moreover, our method produces an SVM classifier that allows linear time classification of test sequences. Our experiments provide evidence that string-based kernels, in conjunction with SVMs, could offer a viable and computationally efficient alternative to other methods of protein classification and homology detection. PMID- 11928509 TI - Detecting positively selected amino acid sites using posterior predictive P values. AB - Identifying positively selected amino acid sites is an important approach for making inference about the function of proteins; an amino acid site that is undergoing positive selection is likely to play a key role in the function of the protein. We present a new Bayesian method for identifying positively selected amino acid sites and apply the method to a data set of hemagglutinin sequences from the Influenza virus. We show that the results of the new methods are in accordance with results obtained using previous methods. More importantly, we also demonstrate how the method can be used for making further inferences about the evolutionary history of the sequences. For example, we demonstrate that sites that are positively selected tend to have a preponderance of conservative amino acid substitutions. PMID- 11928511 TI - A stability based method for discovering structure in clustered data. AB - We present a method for visually and quantitatively assessing the presence of structure in clustered data. The method exploits measurements of the stability of clustering solutions obtained by perturbing the data set. Stability is characterized by the distribution of pairwise similarities between clusterings obtained from sub samples of the data. High pairwise similarities indicate a stable clustering pattern. The method can be used with any clustering algorithm; it provides a means of rationally defining an optimum number of clusters, and can also detect the lack of structure in data. We show results on artificial and microarray data using a hierarchical clustering algorithm. PMID- 11928510 TI - Improving sequence alignments for intrinsically disordered proteins. AB - Here we analyze sequence alignments for intrinsically disordered proteins. For 55 disordered protein families we measure the performance of different scoring matrices and propose one adjusted to disordered regions. An iterative algorithm of realigning sequences and recalculating matrices is designed and tested. For each matrix we also test a wide range of gap penalties. Results show an improvement in the ability to detect and discriminate related disordered proteins whose average sequence identity with the other family members is below 50%. PMID- 11928512 TI - Ab initio folding of multiple-chain proteins. AB - Our previous methodology for ab initio prediction of protein structure is extended here to treat multiple-chain proteins. This involved modification of our united-residue (UNRES) force field and our Conformational Space Annealing (CSA) Global Optimization procedure. Good results have been obtained for both a four- and a three-helix protein from the CASP3 exercise. PMID- 11928514 TI - Using evolutionary methods to study G-protein coupled receptors. AB - A novel method to analyze evolutionary change is presented and its application to the analysis of sequence data is discussed. The investigated method uses phylogenetic trees of related proteins with an evolutionary model in order to gain insight about protein structure and function. The evolutionary model, based on amino acid substitutions, contains adjustable parameters related to amino acid and sequence properties. A maximum likelihood approach is used with a phylogenetic tree to optimize these parameters. The model is applied to a set of Muscarinic receptors, members of the G-protein coupled receptor family. Here we show that the optimized parameters of the model are able to highlight the general structural features of these receptors. PMID- 11928513 TI - Investigating evolutionary lines of least resistance using the inverse protein folding problem. AB - We present a polynomial time algorithm for estimating optimal HP sequences that fold to a specified target protein conformation based on Sun et al's Grand Canonical (GC) model. Application of the algorithm to related proteins taken from the PDB allows us to explore the nature of the protein genotype:phenotype map. Results suggest: (1) that the GC model captures important biological aspects of the mapping between protein sequences and their corresponding structures, and (2) the set of sequences that map to a target structure with optimal energy is affected by minor differences in structure. PMID- 11928515 TI - Progress in predicting protein function from structure: unique features of O glycosidases. AB - The Structural Genomics Initiative promises to deliver between 10,000 and 20,000 new protein structures within the next ten years. One challenge will be to predict the functions of these proteins from their structures. Since the newly solved structures will be enriched in proteins with little sequence identity to those whose structures are known, new methods for predicting function will be required. Here we describe the unique structural characteristics of O glycosidases, enzymes that hydrolyze O-glycosidic bonds between carbohydrates. O glycosidase function has evolved independently many times and enzymes that carry out this function are represented by a large number of different folds. We show that O-glycosidases none-the-less have characteristic structural features that cross sequence and fold families. The electrostatic surfaces of this class of enzymes are particularly distinctive. We also demonstrate that accurate prediction of O-glycosidase function from structure is possible. PMID- 11928516 TI - Support vector machine prediction of signal peptide cleavage site using a new class of kernels for strings. AB - A new class of kernels for strings is introduced. These kernels can be used by any kernel-based data analysis method, including support vector machines (SVM). They are derived from probabilistic models to integrate biologically relevant information. We show how to compute the kernels corresponding to several classical probabilistic models, and illustrate their use by building a SVM for the problem of predicting the cleavage site of signal peptides from the amino acid sequence of a protein. At a given rate of false positive this method retrieves up to 47% more true positives than the classical weight matrix method. PMID- 11928517 TI - Ontology development for a pharmacogenetics knowledge base. AB - Research directed toward discovering how genetic factors influence a patient's response to drugs requires coordination of data produced from laboratory experiments, computational methods, and clinical studies. A public repository of pharmacogenetic data should accelerate progress in the field of pharmacogenetics by organizing and disseminating public datasets. We are developing a pharmacogenetics knowledge base (PharmGKB) to support the storage and retrieval of both experimental data and conceptual knowledge. PharmGKB is an Internet-based resource that integrates complex biological, pharmacological, and clinical data in such a way that researchers can submit their data and users can retrieve information to investigate genotype-phenotype correlations. Successful management of the names, meaning, and organization of concepts used within the system is crucial. We have selected a frame-based knowledge-representation system for development of an ontology of concepts and relationships that represent the domain and that permit storage of experimental data. Preliminary experience shows that the ontology we have developed for gene-sequence data allows us to accept, store, and query data submissions. PMID- 11928518 TI - Constraint-based hydrophobic core construction for protein structure prediction in the face-centered-cubic lattice. AB - We present an algorithm for exact protein structure prediction in the FCC-HP model. This model is a lattice protein model on the face-centered-cubic lattice that models the main force of protein folding, namely the hydrophobic force. The structure prediction for this model can be based on the construction of hydrophobic cores. The main focus of the paper is on an algorithm for constructing maximally and submaximally compact hydrophobic cores of a given size. This algorithm treats core construction as a constraint satisfaction problem (CSP), and the paper describes its constraint model. The algorithm employs symmetry excluding constraint-based search and relies heavily on good upper bounds on the number of contacts. Here, we use and strengthen upper bounds presented earlier. The resulting structure prediction algorithm (including previous work) handles sequences of sizes in the range of real proteins fast, i.e. we predict a first structure often within a few minutes. The algorithm is the first exact one for the FCC, besides full enumeration which is impracticable for chain lengths greater than about 15. We tested the algorithm successfully up to sequence length of 160, which is far beyond the capabilities even of previous heuristic approaches. PMID- 11928519 TI - Detecting native protein folds among large decoy sets with hydrophobic moment profiling. AB - A new hydrophobic score will be presented in this paper for detecting native-like folds from a large set of decoy structures. A recent paper (B. D. Silverman, PNAS 98, 4996, 2001) had revealed that for globular proteins there exists a relatively universal constant of 0.74 for a hydrophobic ratio, which is defined as the ratio of radii from the protein centroid at which the second order hydrophobic moment and the zero order moment vanishes. This paper further defines a new hydrophobic score which will be used to examine protein decoys, in particular, the Holm & Sander, Park & Levitt and Baker decoy sets. It will be shown that the hydrophobic score and profile shapes can provide useful information that should be complementary to the information provided by other procedures, such as free energy calculations. PMID- 11928520 TI - A SOFM approach to predicting HIV drug resistance. AB - The self-organizing feature map (SOFM or SOM) neural network approach has been applied to a number of life sciences problems. In this paper, we apply SOFMs in predicting the resistance of the HIV virus to Saquinavir, an approved protease inhibitor. We show that a SOFM predicts resistance to Saquinavir with reasonable success based solely on the amino acid sequence of the HIV protease mutation. The best single network provided 69% coverage and 68% accuracy. We then combine a number of networks into various majority voting schemes. All of the combinations showed improved performance over the best single network, with an average of 85% coverage and 78% accuracy. Future research objectives are suggested based on these results. PMID- 11928521 TI - Automating data acquisition into ontologies from pharmacogenetics relational data sources using declarative object definitions and XML. AB - Ontologies are useful for organizing large numbers of concepts having complex relationships, such as the breadth of genetic and clinical knowledge in pharmacogenomics. But because ontologies change and knowledge evolves, it is time consuming to maintain stable mappings to external data sources that are in relational format. We propose a method for interfacing ontology models with data acquisition from external relational data sources. This method uses a declarative interface between the ontology and the data source, and this interface is modeled in the ontology and implemented using XML schema. Data is imported from the relational source into the ontology using XML, and data integrity is checked by validating the XML submission with an XML schema. We have implemented this approach in PharmGKB (http://www.pharmgkb.org/), a pharmacogenetics knowledge base. Our goals were to (1) import genetic sequence data, collected in relational format, into the pharmacogenetics ontology, and (2) automate the process of updating the links between the ontology and data acquisition when the ontology changes. We tested our approach by linking PharmGKB with data acquisition from a relational model of genetic sequence information. The ontology subsequently evolved, and we were able to rapidly update our interface with the external data and continue acquiring the data. Similar approaches may be helpful for integrating other heterogeneous information sources in order make the diversity of pharmacogenetics data amenable to computational analysis. PMID- 11928522 TI - Treating tobacco use and dependence. Clinical practice guidelines. PMID- 11928523 TI - Medicaid restraint and seclusion rule. PMID- 11928524 TI - Differential age effects in semantic and episodic memory, Part II: Slope and intercept analyses. AB - This study tested for dissociations between age-related slowing in peripheral and central processes across semantic and episodic memory tasks. We report the mixed model analysis of variance (ANOVA) results using reaction time (RT), error, and data from four different experimental tasks and eight data sets (the four tasks involved either multiple sessions or different stimuli) conducted on the same 80 participants (40 younger and 40 older adults). An earlier paper (Allen, Sliwinski, Bowie, & Madden, in press) reported the factor analysis and structural equation modeling results of this study. We employed two semantic memory tasks (lexical decision and multiplication verification) using data from two different sessions (for a total of four semantic data sets), and two episodic memory tasks (hybrid visual search and memory search with digits and with words as stimuli). For RT analyses, we found Age x Task interactions for all four episodic memory data sets, but no Age x Task interactions for the four semantic memory data sets. Also, the intercept (assumed to index peripheral processes) and slope (assumed to index central processes) analyses demonstrated that older adults (relative to younger adults) showed both intercept and slope decrements for episodic memory tasks, but only intercept decrements for semantic memory tasks. These data are consistent with a theoretical framework in which there are age-related dissociations between peripheral and central processes across semantic and episodic memory. PMID- 11928525 TI - Pathological determinants of the transition to clinical dementia in Alzheimer's disease. AB - The authors developed multivariate models to examine the association between Clinical Dementia Rating (CDR) scale scores and the spatial distribution of paired helical filament tau (PHF-tau) pathology. Severity of tauopathy was examined in 14 cortical regions of interest (ROIs). Classification trees were used to test the independent hierarchical contributions of each ROI to dementia. Multiple-regression and cluster analyses were performed to determine the relative contributions of select ROIs to dementia. Dementia (CDR > or = 1) was modeled as a dichotomous variable. Autopsy material was obtained from 124 demented and nondemented elderly patients. All ROIs except the hippocampus made significant contributions to dementia. However, they were not independent. In multivariate models, only a single step (Step 7 in a hierarchical progression), which contained four ROIs, contributed significantly to dementia. A classification tree resulted in a single decision split, suggesting that only Step 7 ROIs need be considered. A total of 89.4% of the cases were correctly classified (p < .0001). Twelve discrepant cases all had superimposed vasculopathy that might also have affected the function of Step 7 ROIs. The transition to clinical dementia was associated with the presence of tauopathy in A9/10, A22, A23, and A39. Animal studies suggest that these represent a single distributed cortical network focusing on prefrontal regions that provide "executive control" over complex goal directed behaviors. A22, A23, and A39 provide major afferents to other frontal systems and have previously been implicated in very early clinical Alzheimer's disease. PMID- 11928526 TI - Urinary free cortisol is similar in older and younger women. AB - The increased cortisol secretion that occurs with aging is associated with characteristics of frailty. In this study we investigated whether healthy elderly women excrete increased amounts of urinary free cortisol (UFC) when compared to younger women. Twenty-four-hour UFC levels were measured in 25 elderly (70 to 87 years of age; median: 74) and 24 younger (19 to 59 years of age; median: 46) community-living women. All subjects were healthy and cognition was unimpaired. No differences in 24-hr UFC were observed between elderly and younger women (54.2 +/- 28.0 and 45.1 +/- 15.6 micrograms/24 hr, respectively) and no correlation was found between 24-hr UFC and age. Also, elderly and younger women had similar 24 hr urinary volumes (1366 +/- 531 and 1514 +/- 412 ml 24 hr, respectively) that did not correlate with UFC. In conclusion, older and younger healthy women have similar 24-hr UFC levels, possibly reflecting a comparable adrenocortical secretion of cortisol. PMID- 11928528 TI - Informed consent for medical research by the elderly. AB - Much has been written about the possible lack of capacity of the elderly, especially certain vulnerable populations of older individuals, to give informed consent for medical research. Several small studies have shown a deficit for comprehension of consent material, by the elderly, especially those with less education, but this appears small in comparison to an overall deficit in the general population. A number of investigations have suggested that deficits in executive control functions (ECFs) may be related to lack of capacity to make clinical judgments, but these have yet to be applied to research. Many methods have been piloted to measure capacity and to improve comprehension, some of which may help, although none has been proved conclusively to do either. As the elderly experience significant morbidity and mortality from a vast array of illnesses, the use of the elderly as subjects of medical research is especially important. To prevent older individuals from being coerced into participating or potentially being harmed by scientific investigation, they must give informed consent to their involvement. However, there are many studies to suggest that they are not well informed. A discussion ensued in the 1970s and 1980s about whether or not the elderly deserved special protection as a class of individuals, based on their possible increased risk during medical experimentation, but it was ultimately decided that, because the majority of elderly are perceived to be cognitively intact, they need not receive additional safeguards (High & Doole, 1995, Behavioral Science and Law, 13, 319-335). The U.S. Department of Health and Human Services in 2000 (Federal Registrar Rules and Regulations, 46, 8366-8392) reviewed existing protections for subjects of human research and deemed they were inadequate, issuing new guidelines. This article reviews evidence that differences exist between the ability of young and old in their capacity to give consent. Alterations in methods of obtaining consent may help individuals to give a more informed consent, and even enable subjects who lack the capacity to consent, such as cognitively impaired individuals, to participate in research. However an ideal means of screening or altering the consent process has yet to be devised. Many of these methods are briefly considered. PMID- 11928527 TI - The effects of age-stereotype priming on the memory performance of older adults. AB - The effects of age-stereotype priming on the memory performance of older adults were investigated through a conceptual replication and extension of Levy's (1996. Journal of Personality and Social Psychology, 71, 1092-1107) study. Sixty young and 60 older adults were subliminally primed with a positive age stereotype, a negative age stereotype, or neutral primes. Memory performance on two tasks (a photo recall task and a dot location task) was measured before and after the priming intervention. Although the study does not provide unequivocal support for Levy's (1996) findings, results did show that priming a negative age stereotype undermined memory performance for a small sample of older adults who were "unaware" of the primes. However, contrary to Levy's findings, priming a positive age stereotype did not increase older adults' memory performance. There were no significant effects of priming positive or negative age stereotypes on the memory performance of young adults. PMID- 11928529 TI - Recall and subjective reactions to speaking styles: does age matter? AB - Eighty-four younger adults (M = 20.9 years), and 70 older adults (M = 71.1 years) watched a videotaped presentation of medication instructions presented in either neutral speech or elderspeak. Older adults, particularly those with higher working memory performance, tended to recall more information from the elderspeak version. Younger and older adults agreed in rating the elderspeak as having both positive and negative characteristics. Findings supported our hypothesis that the relationship between recall performance and positive subjective reactions to speaking styles would be stronger for older adults than for younger adults. PMID- 11928530 TI - Age-related changes in the kinematics of curved drawing movements: relationships between tangential velocity and the radius of curvature. AB - Age-related motor slowing may result from abnormal trajectory formation. Curved trajectories can be described by a 1/3 power law linking velocity and radius of curvature. As a basic coordinative principle that emerges with maturation, factors producing any variation in this relationship may thus provide insights into age-related changes in the coordination of movement. The authors determined whether variations in the fit of a 1/3 power law with advancing age were associated with temporal or postural constraints. Young and older adults moved under temporal (speed and timing cues) or postural (joint combinations used) constraints, while performing elliptical movements upon a graphics tablet at 1 or 2 Hz. A 1/3 power law was influenced by variations in speed and the joint combinations employed, but reductions of fit were not necessarily linked to force inefficiency. Age-related reductions in the strength of a 1/3 power law were a function of postural constraints. PMID- 11928531 TI - [Challenges in the treatment of patients with type 2 diabetes mellitus and obesity]. AB - The current treatment of obesity generally fails in the long term and the optimized control of diabetes represents a difficult challenge requiring a multi disciplinary approach and a high degree of motivation by the therapeutic team, the patient and the patient's family. When diabetes is associated with obesity, quite a frequent phenomenon, the treatment of these patients represents a true clinical challenge. Although far from perfect, there are currently several therapeutic weapons available in the armoury. Non-pharmacological treatment, fundamentally a low calorie diet and its multiple variations, anti-glycaemia and anti-obesity drugs, and surgical treatment or a combination of these therapeutic strategies present various difficulties and pose several questions. The characteristics of each are analyzed with their pros and cons and their implication in weight loss, control of glycaemia, reduction in the associated cardiovascular risk factors, morbidity and mortality. PMID- 11928532 TI - [Fiber in the treatment of obesity and its comorbidities]. AB - A considerable part of the health-care significance of obesity lies in its co morbidity. Obesity is associated with high blood pressure, type 2 diabetes mellitus and dyslipidaemia, representing both individually and when taken together risk factors for cardiovascular disease, the main cause of death in industrialized countries. The influence of fibre consumption on body weight, the risk of type 2 diabetes and cardiovascular disease has been shown in important epidemiological surveys, but the mechanisms whereby fibre exercises this protective effect have not been definitively clarified. For the most part, the ability of fibre to slow down the process of absorption of nutrients by modifying the metabolic response to intake, its ability to increase the excretion of biliary acid and the metabolic effects of the short-chain fatty acids produced in the bacterial fermentation of fibre have all focussed the attention of the working hypotheses analyzed in the studies published in the last few years. This review is intended to identify the importance of fibre for the prevention and treatment of obesity and its comorbidities on the basis of the available evidence at present. PMID- 11928533 TI - [Genetic polymorphisms and new perspectives in the prevention and treatment of obesity]. AB - The prevalence of obesity has increased to an extraordinary degree, especially over the last three decades, so that if these trends persist, practically all of the adult population would be obese in the course of only two generations. On the basis of family and adoption studies, it has been estimated that the genetic component in obesity ranges from 20% to 80%. Thus, the most common forms of obesity would depend on the interaction of multiple genes as well as on the influence of various environmental factors, such as eating behaviour and lifestyle. Although none of these genes potentially involved in the control of body weight seems to be directly responsible for the syndrome, there have been reports of interesting associations between polymorphisms of certain candidate genes and obesity or its metabolic complications. The studies into associations between genotypes and obese phenotype have increased over the last few years and have basically focussed on the genes involved in the control of the energy balance, giving rise to a whole series of results which might constitute the basis for extremely interesting strategies for the prevention and treatment of this serious problem. PMID- 11928534 TI - [Prevalence of obesity in developed countries: current status and perspectives]. AB - Obesity is one of the main public health problems in developed countries. It is considered to be a risk factor associated with the genesis or development of the major chronic diseases. The classification of a population by weight and the awareness of its determining factors are essential elements in order to undertake health planning programmes and health promotion projects. In Europe, the prevalence of obesity (BMI 30) is greater in northern countries (7% in men and 9% in women in the Netherlands; 5.3% and 9.1% respectively in Sweden) than in southern countries (Spain: 11.5% in men and 15.2% for women) or further east (Germany: 10.6% in men and 11.6% among women). In the United States, obesity is taking on alarming proportions, with an increase of 8 percentage points between 1976 and 1994, placing the problem at a level of 22.5% of the population classified as obese. The prevalence of obesity in Spain is 13.4%, higher after the age of 45 years, among women and in individuals with a lower educational level. The pattern of geographical distribution evidences a greater prevalence towards the south and south-east, with statistical significance among males. It is necessary to analyze in further depth the factors involved in the genesis of obesity and to institutionalize community health programmes for its prevention, monitoring and treatment. PMID- 11928535 TI - [Leptin in the regulation of energy balance]. AB - Leptin, identified in 1994 as a substance synthesized in adipose tissue in amounts that are proportional to the magnitude of fat reserves, has been described as an important factor in the regulation of the body's energy homeostasis as it is capable of inhibiting intake, stimulating energy expenditure and regulating other peripheral metabolic processes involved in the control of the spread of body fat (secretion of insulin, lipolysis, glucose transport). Nonetheless, leptin is a pleotropic substance also involved in fertility, inflammation and angiogenesis processes, among others. Although these actions have been well defined in certain animal models of obesity, the role played by leptin in humans is less clear since, at least at the hypothalamic level, there seems to be considerable resistance to the effects of this hormone. The trials which administered recombinant leptin to humans recently have not provided sufficiently hopeful results as the weight reductions obtained have not been greater than those achieved with other drugs currently available on the market and its subcutaneous administration has led to some undesirable side effects. In any case, the results obtained must be treated with caution, with further trials being needed on the biological activity of leptin in humans in order to determine the clinical repercussions of the administration of this hormone. PMID- 11928536 TI - [Obesity and osteoporosis: effect of weight variation on bone mass]. AB - Obesity and osteoporosis are both chronic conditions with multi-factor aetiologies and a considerable associated level of morbidity and mortality. They are endemic in economically developed countries. Various epidemiological studies have indicated an overweight condition to have a protective effect on the occurrence of osteoporotic fractures, but the data available on the effects of weight loss on the conservation of bone mass are scant and fragmented. The present review attempts to give a critical assessment of the published trials that have analyzed the evolution of bone mass during the administration of low calorie diets to obese females, as well as to identify those aspects that would deserve special attention in the planning of future research in this field. PMID- 11928537 TI - [Binge eating disorders and obesity]. AB - The present article reviews binge eating disorders. These comprise a set of eating behaviour alterations that are characterized by the persistent intake of large amounts of food during discrete periods of time, with moderate levels of associated psychopathology, and they are quite resistant to treatment. Of all these, binge eating or compulsive eating disorder (TA in its Spanish acronym) is the one most recently identified and, therefore, the one requiring the most additional investigation. Although between 20% and 30% of patients with TA are obese, the nature of the relationship between these two disorders is not known even today. In the same way, there are insufficient empirical data demonstrating the role of following diets in the aetiology of TA and the repercussions of restriction in the treatment of TA. For the time being, the combination of cognitive behavioural therapy and behavioural strategies for weight loss is the most effective procedure to reduce binge eating and to facilitate short-term weight loss; the addition of anti-depressive medication or appetite suppressants seems to give a moderate increase in weight loss. Finally, there is evidence that cognitive-behavioural self-help procedures without therapeutic contact or with a minimal therapeutic contact may be useful in obese patients with TA. PMID- 11928539 TI - [Role of lipid intake in obesity]. AB - It is a commonly accepted fact that the fat in our diet plays an important role in the onset and maintenance of obesity. The excess consumption of energy associated with a high intake of fat and the greater metabolic efficiency in its use are the mechanisms suggested to support the relationship between dietary fat and adiposity. Nonetheless, the epidemiological evidence in favour of lipid intake as a promoter of obesity is not conclusive. Intervention studies, on the other hand, consistently show that there is a modest decrease in weight associated with low fat diets ad libitum, which seems to be explained by the reduction in the intake associated with such an intervention. However, this effect on weight is not maintained over time as considerable reductions in the intake of lipids in the long term seem to have no or minimal effects on corporal adiposity. It has been consistently proven that the restriction of total calories leads to greater weight losses than those achieved exclusively with fat restrictions and, on the other hand, there is very little evidence that low-fat diets per se lead to a weight loss regardless of the calorie restriction. Finally, it must be remembered that low-fat diets rich in carbohydrates are not without undesirable side effects, particularly on cardiovascular risk factors. In conclusion, a change in habits leading to an overall restriction in calories and the promotion of physical activity is a much more desirable strategy in the treatment and prevention of obesity than the apparently promising restriction of lipids in isolation. PMID- 11928540 TI - [Benefits, efficacy, and risk associated with the conventional treatment of obesity: should we treat the weight excess?]. AB - The literature contains a large number of published articles indicating a strong relationship between obesity and morbidity-mortality due to different cause. There are also various short- to medium-term studies demonstrating the benefits of a moderate weight loss on the control of obesity-related risk factors. Nonetheless, very few studies have attempted to define the potential medium- to long-term morbidity/mortality benefits associated with weight loss using conventional obesity treatment methods. This review attempts to weigh up the benefits and also the risks associated with the methods currently recognized for the treatment of this pathology. We also attempt to analyze the efficacy of these methods for controlling body weight in the medium and long term, as this is essential when it comes to advising the obese population. PMID- 11928538 TI - [Insulin resistance and evolution]. AB - Both infections and injuries activate the immunity system, leading to a series of metabolic changes which place the organism at a disadvantage and contribute to its elimination, thus facilitating the repair of the injured tissue. The study of the actions of tumour necrosing factor alpha (TNF-alpha) and interleukin-6 (IL 6), classically implicated in inflammatory processes and in fighting infection, has revealed numerous metabolic effects. Some gene polymorphisms of TNF-alpha and IL-6 (associated with a different TNF-alpha or IL-6 transcription rate) and the plasma concentrations of the soluble TNF-alpha receptor are found to be simultaneously associated with resistance to insulin, the proportion of body fat and with the mortality linked with different chronic infections. Therefore, it seems that the immune system is designed to fight infections effectively and to provide certain survival advantages during periods of intermittent fasting so frequent in the past. By inducing a resistance to insulin in the muscles, the energy substrates would thus be reserved for neuronal metabolism. In the presence of an insulin-resistance genotype and a westernization of the environment (carbohydrate-rich diet, an increase in saturated fat, low fibre and sedentary lifestyle), a genotype with a high cytokine response will contribute to a worsening of the resistance to insulin and, finally, to type 2 diabetes mellitus and atherosclerosis. The advantages for our ancestors of a large cytokine response (eradication of the lesion) or moderate resistance to insulin (protection against food shortage) have led in the present day to the development of atherosclerosis now that the characteristics of the environment have changed. It is contended that these changes constitute examples of good adaptation to the environment or poor concordance between our current lifestyle and our genome. PMID- 11928541 TI - Sexual science in ancient Greece. PMID- 11928542 TI - The reluctant acceptance of new ideas in medicine. PMID- 11928543 TI - Reflections on the lost art of caring. PMID- 11928544 TI - Leaders in American Medicine. David C. Sabiston, Jr., M.D. PMID- 11928545 TI - Diesel gas, rice, and medical errors. PMID- 11928547 TI - [The long-term evaluation of artificial myoplastic anus and artificial classical anus based on manometric and radiological examinations]. AB - The aim of this study was comparative evaluation of the pressure profile in the artificial myoplastic anus and the one created by the use of classical method. Radiological examinations of the colon by the use of the contrasting agent were performed in all patients in each group. In the group of patients with artificial myoplastic anus radiological examinations showed the narrowing of the lumen 4-7 cm in length in all patients. Above the narrowing the widening of the lumen was present. Reflexive flow of the contrasting agent from the colon was not present. In the group of patients with the artificial classical anus the narrowing of the terminal part of the colon was not noticed in the radiological examination. In all patients the reverse flow of the contrasting agent through the artificial anus was observed. Patients with the artificial myoplastic anus created by the use of pedunculated fold of the muscular coat of the colon have much higher average pressures around the myoplastic sphincter than in the presphincteric area, while the patients with the artificial classical anus did not present the area with the increased pressure in the terminal part of the colon. The possibility of preventing uncontrolled defecation and flatulence by the patients with the artificial myoplastic anus is related to the repeatedly higher pressures around the sphincter than in the presphincteric area, which do not decrease as the time passes by after the surgery. PMID- 11928548 TI - [Concentration of glutathione (GSH), ascorbic acid (Vit. C), and thiobarbiturate acid reacting components (MDA) in brain neoplasms]. AB - The aim of the study was to evaluate the concentration of glutathione (GSH), ascorbic acid (Vit C), and thiobarbiturate acid reacting components (MDA) in brain neoplasms in specimens from normal brain tissue. The group of 72 individuals treated surgically for brain neoplasm in Department of Neurosurgery Medical Academy of Bialystok (Poland) in the period from 1996 to 1999 was included into the study. The GSH concentration was estimated with GSH-400 method, ascorbic acid by the use of Kyaw method, and MDA by Salaris and Babs method. The statistical analysis revealed diminished concentration of GSH and Vit. C (p < 0.001), and analogous increase of MDA concentration (p < 0.001) in the investigated specimens compared to the mentioned above substances concentration in the specimens obtained from normal brain tissue. PMID- 11928549 TI - [Laparoscopic varicocele ligation. The comparative assessment of artery-ligating and artery-preserving varicocelectomy]. AB - For over ten years more and more significant role in the diagnostics and clinical treatment of marital infertility have been ascribed to the male factor, which accounts for 30-40% of infertility cases. One of the most common reasons of male infertility is varicocele. Between the years 1994-2000 transperitoneal laparoscopic varicocelectomy was performed in 166 men with unilateral varicoceles. Two methods of surgical correction were administered. In I group--88 men (53%)--testicular artery and vein were ligated, in II group--78 men (47%) only testicular vein was ligated. The success of the treatment was evaluated in two age groups; A--19-30 and B--31-41 years of age. Before the operation in group I and II the sperm count was respectively 18.19?10.51 and 19.05 +/- 10.62 x 10(6)/ml. The percentage of the progressive motility was 18.95 +/- 9.46% in I group and 20.12 +/- 9.08% in II group. The percentage of normal forms was 22.04 +/- 8.42% in I group and 23.02 +/- 9.81% in II group. After 12 months of surgical observation the sperm count was 35.21 +/- 13.46 in I and 31.82 +/- 12.14 x 10(6)/ml, in II group. The percentage of progressive motility increased to 36.48 +/- 15.76% in I group and to 31.26 +/- 10.78% in II group. In both groups the percentage of normal forms was comparable, in I group was 49.11 +/- 26.72% and in II group was 47.15 +/- 17.18%. In both age groups sperm count before operation was respectively: 20.42 +/- 10.20 in A, and 16.42 +/- 9.16 x 106/ml in B. The percentage of progressive motility was in A and B respectively 22.98 +/- 9.91% and 15.38 +/- 9.66%. At percentage of normal forms was respectively 25.62 +/- 8.92 and 18.81 +/- 7.93%. In the end of observation period the improvement of sperm count was observed in A--42.69 +/- 22.36, in B--22.86 +/- 11.76 x 10(6)/ml. The improvement of progressive motility was ia A--43.27 +/- 23.96% and in B- 23.09 +/- 17.01%. The improvement of normal forms were in A--55.39 +/- 24.31% and 39.69 +/- 22.29 in group B. The pregnancy rate was; in I group +/- 30.6% in II group--23.1%, in A group--33.3%, in B--group--19.7%. The laparoscopic ligation of the testicular vein and artery produced better improvement of semen characteristics and percentage of pregnancies in comparison with artery sparing technique. The laparoscopic varicocelectomy regardless of the technique allows a significant improvement of semen characteristics and pregnancy rate among the couples, where men were less than 30 years of age. PMID- 11928550 TI - [Coronary artery bypass grafting in patients over 70 years of age]. AB - The purpose of this study was to assess the risk of coronary artery bypass grafting (CABG) in the group of patients (pts) over 70 years of age and to examine results of surgical treatment in these pts. Between January 1993 to December 1999 1276 pts underwent CABG. From this group 114 consecutive pts at the age over 70 were retrospectively studied. The average age of the pts was 71.8 +/- 2.2 years (range, 70 to 79 years). Eighty nine male (78.1%) and 25 female (21.9%) pts were operated. One or more myocardial infarction had occurred in 64.0% of pts preoperatively. All the pts underwent left cardiac catheterization. The indication for surgery was significant stenosis of the left main trunk in 35 (30.7%) pts and three vessels disease in 54 (47.4%) pts. The total number of peripheral anastomoses was 274 (average 2.4 +/- 0.9 grafts per pts). In 29 cases left internal mammary artery (to left anterior descending artery) and in 245 reversed saphenous vein grafts were used. The early postoperative mortality in the group of pts at the age over 70 (10 pts--8.8%) was significantly higher than in the group of younger pts (41 of 1162 pts). The cause of death in septuagenarians was: low cardiac output syndrome (5 pts), respiratory failure (2 pts), renal insufficiency (2 pts) and cerebral stroke (1 pt). In early postoperative course the incidence of any organ insufficiency (cardiac, respiratory, renal and neurological) was higher in the group of pts over 70 years of age than in the younger pts. Seven-year probability of survival calculated from Kaplan-Meier method was 66 +/- 4%. After operation 84.5% of pts were asymptomatic, while only 5 pts were in III and IV CCS functional class. CABG in pts at the age over 70 is associated with higher operative risk and higher rate of perioperative organ failure. After surgery most pts enjoy improvement in life quality. PMID- 11928551 TI - [The physical capacity of men with ischaemic heart disease after cardiac rehabilitation]. AB - The aim of the study was the evaluation of physical capacity of cardiovascular system and the ability to perform permitted exercises in a group of 114 patients who underwent pharmacological or surgical treatment of ischaemic heart disease and subsequently were submitted to 12 month lasting cardiac rehabilitation. Rehabilitation programme comprised hospital, sanatorium and ambulatory periods. The control group comprised 29 patients without changes in coronary arteries. All patients underwent treadmill test, according to Bruce protocol. The efficacy of rehabilitation was estimated in the particular groups regarding to the method of treatment. 143 men, were enrolled in the study in following groups: A--patients with the history of myocardial infarction (MI) treated pharmacologically (n = 29) B--patients with the history of MI after surgical treatment (n = 29) C--patients without MI after surgical treatment (n = 28) D--patients without MI treated pharmacologically (n = 28) K--control group (n = 29). All men in groups A, B, C completed cardiac rehabilitation programme. The highest level of physical capacity was observed in group of operated (CABG) patients. Patients treated pharmacologically reached significantly lower level of physical capacity. PMID- 11928552 TI - [Exercise tolerance in patients after acute lymphoblastic leukemia treatment in childhood]. AB - The purpose of the presented study was to define the exercise tolerance in patients after acute lymphoblastic leukemia (ALL) treatment in childhood. Three groups of persons were examined: group A: 20 children, aged 7-19 years (mean 12.4 y), examined immediately after ALL therapy completion, with cumulative anthracycline (ATC) doses administered 155.8-300 mg/m2 and dexrazoxane, as cardioprotectant, group B: 36 patients, aged 12-24 years (mean 15.9), being 3-5 years after ALL treatment, who received ATC in cumulative doses 148.6-416.7 mg/m2, without cardioprotection, group C: 28 healthy volunteers, aged 9-25 years (mean 17.3), as controls. All the examined patients belonged to NYHA functional class I. In all subjects the exercise treadmill test was performed according to modified Bruce protocol. The parameters analysed were: MET--number of metabolic effort units achieved at the test, HRmax--maximal heart rate during exercise, %HRmax--percent of maximal HR for given patient's age achieved during the STdep- depression of ST segment in electrocardiography (ECG) immediately after the maximal exercise. During the exercise members of all 3 groups achieved the required HRmax without serious complaints and ECG abnormalities. Examined persons in group A,B and C presented with effort levels (MET), %HRmax, STdep that did not differ significantly. Only HRmax in groups A and C were higher than that achieved by members of group B. PMID- 11928553 TI - Function of insulin-like growth factor (IGF-I) and its binding protein (IGFBP-1) in pathological proliferation of endometrium. AB - The purpose of this project was the investigation of insulin, IGF-I and IGFBP-1 concentrations in the serum of patients with pathological changes in endometrium. IGF-I and its binding protein (IGFBP-1) stimulate cell growth and differentiation of the normal and neoplastic endometrium. The mean values of IGF-I concentrations in the group of patients with atypical hyperplasia and endometrial carcinoma were significantly higher than in the control group with normal endometrium. Significantly higher IGFBP-1 concentrations were found in the serum of patients with the atypical hyperplasia of endometrium and endometrial carcinoma. The correlation between insulin concentration and changes in endometrium in the examined groups was not found. PMID- 11928554 TI - [Misoprostol and oxytocin in induction of labour in women with prolonged pregnancy--safety and effectiveness comparison]. AB - 16 women with prolonged pregnancy and indications to induction of labour were involved into prospective analysis. Sixty of them were given 50 micrograms of Mizoprostol straight into posterior vaginal fornix (only once). In 56 women the labour was induced by intravenous administration of 5 IU Oxytocin. Effectiveness and safety of both preparations used in labour induction were compared. Non significant higher effectiveness and significant faster induction of uterine contractions by Oxytocin was found. Mizoprostol administration significantly shortens duration of the first labour period. Eventually, time from the medicine administration until the labour in both groups was comparative. PMID- 11928555 TI - [Estimation of platelet counts and their morphological parameters in patients infected by borrelia burgdorferi]. AB - Platelet counts and their morphologic parameters in patients with Lyme borreliosis before and after antibiotic therapy (4 weeks of treatment) were estimated. 30 patients aged 17-60, x = 41 were evaluated: 7 with Erythema migrans, 3 patients with neuroborreliosis in the from Lymphocytic meningitis and 20 ones with Lyme arthritis. Control group consisted of 19 healthy persons aged 34-52, x = 43. Hematologic analyser Coulter MAXM was used for testing PLT, MPV, PCT and PDW. The results indicated that patients with Lyme boreliosis have decreasing platelet count with simultaneously increasing their volume in comparison with healthy control. It may result from the involement of platelets in defense mechanisms of infected host. The decrease of platelet count after the antibiotic treatment in comparison with the control group may be the reflection of influence of antibiotic treatment on thrombopoesis. PMID- 11928557 TI - [Ischaemic heart disease and chronic helicobacter pylori infection--present views]. AB - In connexion with the high prevalence of coronary heart disease its additional reasons are searched, which could lead to the occurrence and development of the disease. Chronic infections have been lately recognized as an additional reason. In the article we presented opinions regarding the influence of long-lasting Helicobacter pylori infection on the development of inflammation process, atheromatous changes, initiation and development of ischaemic heart disease. PMID- 11928556 TI - [The effect of improved glycemic control on quality of life in patients with type I and type II diabetes]. AB - Patients with diabetes (type 1 or type 2) live shorter because of the long-term complications of hyperglycemia. One of the most important aims of therapy in diabetic subjects is delay in the development of long-term diabetes complications. However, another important aim of therapy in diabetic patients is the use of drugs, which not only delay the time of long-term complications development but also improve the quality of life. The paper presents the results of studies on quality of life in diabetic subjects basing on contemporary literature. In patients with type 1 diabetes mellitus a quality of life decreases gradually with the duration of diabetes. The influence of intensive insulin therapy on quality of life is minimal or absent. In UKPDS study no differences in quality of live between conventionally or intensively treated patients were observed. In subjects with macrovascular complications the quality of life is significantly worse. Generally, the authors of UKPDS suggested that better glycemic control is not associated with better quality of life. Only Testa and Simonsen demonstrated that type 2 diabetic subjects treated by new "intelligent" drug--glipizide GITS (Glibenese GITS) is associated not only with better metabolic control but also with better quality of life. PMID- 11928558 TI - [Dehydroepiandrosterone (DHEA)--youth hormone?]. AB - Dehydroepiandrosterone (DHEA) and its sulphated metabolite (DHEA-S) are endogenous steroid hormones, synthesized by the adrenal cortex, gonads and CNS. The secretion profile changes with age and depends on the sex. Human DHEA and DHEA-S levels decline linearly and systematically with age and suggest the potential importance of that parameter as a biomarker of ageing. The counteraction of DHEA against atherosclerotic disease, cancer growth, diabetes mellitus, insulin resistance, obesity and the influence on immunological functions are observed in researches. DHEA influences the condition of mind, cognition functions, memory and well-being. DHEA hormonal replacement therapy is expected to lengthen human life by the stoppage of physiological degeneration changes and prevention of age-related clinical disorders. PMID- 11928560 TI - [Prader-Willi syndrome in 22-year-old man--case study]. AB - Prader-Willi syndrome is a rare genetic disorder with characteristic neonatal hypotonia, followed by obesity, low height, hypogonadism and mental retardation. In this paper a case of 22-year-old man suffering from Prader-Willi syndrome is described. PMID- 11928559 TI - [Management and outcome of pregnancy in women with hypothyroidism--case report]. AB - We analysed the influence of congenital hypothyroidism, diagnosed in the third year of life, on the outcome of pregnancy. We observed the physiological progress of pregnancy and normal development of the foetus. We delivered newborn by caesarean section. We did not observe any disadvantageous influence of congenital hypothyroidism in the mother on newborn status. We analysed the pharmacological substitution and its influence on pregnancy outcome. PMID- 11928561 TI - [Clinical picture of early congenital syphilis in 2,5-month-old infant]. AB - The case of congenital syphilis in a 2.5-month-old infant was describe in the article. Missing information about possibility of infection caused severe course of the disease the and a long-term of treatment. PMID- 11928562 TI - [Lateral cervical cyst in 13-year-old girl]. AB - We present a clinical description of a 13-year-old girl with lateral cervical cyst. We particularly emphasize the need for ultrasonographic examination during routine diagnostics of so-called cervical tumors. PMID- 11928563 TI - [Outstanding contribution of prof. Henryk M. Wisniewski (1931-1999) to the world neuropathology in 20th century]. AB - Born in Poland in 1931, Henryk Miroslaw Wisniewski, obtained MD at the Medical School in Gdansk (1955), where he continued his neuropathological research awarded with Ph. D. in 1960. During 1961-1962 a worked as a Visiting Scientist at NIH (Institute of Neurology and Communicative Diseases and Stroke). In Medical School in Warsaw he was promoted to Docent degree (an associate professor). In 1966 he emigrated with his family to New York, where he was a Research Associate and Professor at Albert Einstein College of Medicine (1966-1975) Subsequently he became a Director of the State Institute for Basic Research in Developmental Disabilities Staten Island. In New York he remained till his early death at the age of 68. Prof. Wisniewski advanced pathological research concerning the development of dementia, including Alzheimer disease. His investigations proved that presentile dementia (Alzheimer disease) is almost identical with senile dementia. That is why he is called the pioneer of modern Alzheimers research. The comments about his scientific contribution were generously published in scientific journals and daily press. The New York Times cited Dr Mony de Leon Prof. of Psychiatry statement reflecting so well Prof. Wisniewski's achievements "He taught us what the lesions for Alzheimers looked like, what they were made of and how they worked". PMID- 11928564 TI - [Methylenetetrahydrofolate reductase gene polymorphism in patients with type 2 diabetes]. AB - Hyperhomocysteinemia is recognised as a risk factor of ischaemic heart disease and vascular complications of arterial hypertension. Methylenetetrahydrofolate reductase (MTHFR) gene C677T polymorphism is associated with hyperhomocysteinaemia. The aim of the study was the assessment of an association of the above polymorphism with type 2 diabetes with special attention to myocardial infarction and arterial hypertension accompanying diabetes. The study group consisted of 172 type 2 diabetics. 172 control subjects with normal glucose tolerance were age and sex matched to patients with diabetes. C677T polymorphism in MTHFR gene locus was detected using polymerase chain reaction restriction fragment length polymorphism (PCR-RFLP) method. CT and TT genotypes were found more often among diabetics (OR 1.83, 95% CI 1.16-2.89; p < 0.01). This finding may be secondary to the excess of T allele bearers among diabetics with myocardial infarction when compared to diabetics without infarction and to control group. Upon obtained results the potential role of genotypes CT and TT as risk factors of myocardial infarction among patients with type 2 diabetes could not be excluded (OR 2.33, 95% CI 0.93-5.8; p = 0.07). Genotypes containing T allele are not associated with diabetes type 2 and concomitant arterial hypertension (OR 1.45, 95% CI 0.89-2.57; p = 0.14). A confirmation in further studies is needed for the presented findings. PMID- 11928565 TI - [Fibrinogen (Fb) concentrations in patients with ischaemic heart disease undergoing coronary angiography]. AB - Increasing evidence suggests the role of hemostatic risk factors in the development of ischemic heart disease (IHD). A raised plasma fibrinogen has been related to increased risk of IHD. The aim of the study was to determine the relationship between plasma fibrinogen and the coronary vessels state based on the coronary angiogram. 119 patients undergoing coronary angiography were classified into 5 groups according the severity of IHD: Group 0 without significant atherosclerotic lesions (control group), Group 1 with single vessel disease, Groups 2, 3 with multivessel disease (two and three affected arteries, respectively) and Group 4 with positive history of myocardial infarction. A statistically nonsignificant rise in fibrinogen levels in Groups 1, 2, 3 (3.9 +/- 0.8 g/l, 4.0 +/- 0.9 g/l, 4.1 +/- 0.9 g/l, respectively) as compared to control Group 0 (3.7 +/- 0.7 g/l) was found. In Group 4 plasma fibrinogen was significantly lower (2.8 +/- 0.6 g/l) comparing to Group 0 (p < 0.05). In addition plasma fibrinogen was positively correlated with blood pressure. These results supports the role of raised plasma fibrinogen in the pathogenesis and development of IHD. PMID- 11928566 TI - [Effects of thrombolysis on left and right ventricular diastolic function in patients after myocardial infraction. One-year follow-up]. AB - It has been shown, that successful reperfusion of the infarctrelated artery by thrombolysis can prevent left ventricular (LV) dilatation and dysfunction. To date no study has shown the impact of thrombolytic therapy on LV and RV diastolic filling. The aim of the study was to assess the effects of thrombolysis on LV and RV diastolic inflow and systolic function of LV in patients (pts) after anterior myocardial infarction MI. METHODS: Echocardiographic and Doppler-derived measurements of LV systolic function, RV and LV diameters and diastolic function at rest and after treadmill exercise test (TET) were assessed prospectively in 21 patients (pts) after MI treated with streptokinase and 10 pts treated without thrombolysis during one-year follow-up. They were compared with 31 age-matched controls. RESULTS: LV and RV filling indices in both groups of pts after MI were impaired compared to controls but in pts without thrombolysis were more abnormal compared to pts treated with thrombolysis. Among several echocardiographic variables most consistent alteration was shortening of E-deceleration time of mitral as well as RV inflow, more abnormal in pts without thrombolysis. There was strong inverse relation between degree of impairment of diastolic variables and exercise capacity. CONCLUSIONS: These data indicates beneficial effect of thrombolysis on RV and LV diastolic function in patients after AMI. E deceleration was the most sensitive marker of diastolic dysfunction. PMID- 11928567 TI - [The effect of inhaled heparin on airway responsiveness to metacholine in asthmatic children]. AB - Atopic asthma is a chronic inflammatory disorder of the airways associated with airway hyperresponsiveness to various bronchoconstrictor stimuli. There are several pharmacologic agents, involved in the inflammatory process, that can produce acute bronchoconstriction, such as metacholine. Heparin possesses multiple noncoagulant properties, including anti-inflammatory actions. OBJECTIVES: The purpose of the present study was to investigate the effect of inhaled heparin on the airway response to metacholine in children with asthma. METHODS: Fifteen children with mild atopic asthma participated in the study. It was randomized, double-blind, cross-over, placebo controlled study. There were three study visits. At the first visit patients were included into the study and beta-agonists were withheld for at least 12 hours before the next visit (second day). At the second visit provocation test with metacholine, inhalation with single dose of heparin or placebo followed by provocation test with metacholine were performed. After two weeks, tests were performed in the same manner except of that patients instead of heparin inhaled placebo or opposite. RESULTS: Fourteen children completed the study. Single dose of inhaled heparin significantly decreased bronchial hyperresponsiveness to metacholine when compared to placebo. CONCLUSION: The exact mechanism of anti-inflammatory role of heparin is still under investigation and needs to be studied further. The possible anti-inflammatory effect of heparin can be through effect on the neuronal M2 receptors or on receptor-protein G binding or direct effect on the smooth muscles, and in such mechanisms, heparin can decrease bronchial hyperreactivity to metacholine. PMID- 11928568 TI - [Evaluation of blood morphology in patients with refractory multiple myeloma treated with thalidomide]. AB - Thalidomide, a derivative of alpha-N-phthalimidoglutarimide acid, was withdrawn from the market in the 1960s because of severe birth defects. Recent reports have suggested antiangiogenic and antitumor activity of this drug. We have treated 52 patients with refractory multiple myeloma at age from 32 to 79 years (mean 63) with thalidomide at a dose of 200-400 mg daily. Out of the group of 52 patients, 27 patients (52%) responded to the therapy, in 25 patients (48%) a response was not achieved (decline in monoclonal protein was smaller than 25%). There was a systematic improvement in haemoglobin concentration, erythrocyte count and thrombocyte count during thalidomide therapy. Leukocyte count showed an inclination to decrease, however observed changes were not statistically significant. The improvement in morphotic parameters of blood was observed both in responder and nonresponder patients. PMID- 11928569 TI - [Analysis of risk factors of the paroxysmal atrial fibrillation after valve replacement in patients with aortic stenosis]. AB - Paroxysmal atrial fibrillation (PAF) is a frequent complication (10-60% of pts) after cardiac surgery. In our study we analyze the influence of clinical, echocardiographic and 24 h ecg parameters on the risk of postoperative PAF in 266 pts with aortic stenosis (88 women and 178 men; mean age 58 +/- 10). PAF was observed in 74 (28%) patients. Statistically significant factors of risk of PAF were (univariate analysis): age-relative risk 1.08 (1.04-1.11), history of PAF- 4.3 (1.4-12.5), more than 100 supraventricular ectopic beats during 24 h ecg--2.9 (1.6-5.1), presence of SVT during 24 h ecg--2.6 (1.5-4.5) and presence of SVT > 140/min--relative risk 3.5 (1.8-6.7). Left atrium diameter and coronary artery bypass grafting during valve replacement had no impact on the risk of PAF. In multivariate analysis three factors remained significant--age, history of PAF and presence of SVT > 140/min during 24 h ecg. In discriminant analysis this model of 3 factors enabled the correct risk assessment in 72% of patients. CONCLUSIONS: 1. The factors that increase the risk of postoperative PAF in pts with aortic stenosis are: age, history of PAF and presence of SVT > 140/min during preoperative 24 h ecg. 2. Postoperative PAF is not related to left atrium diameter in this group of patients. 3. Coronary artery bypass grafting during aortic valve replacement does not increase the risk of PAF. PMID- 11928570 TI - [Granulocytopenia in patient with congenital asplenia and connective tissue disease. Case report]. AB - We are presenting a case of 29-year-old woman with neutropenia, in whom congenital asplenia was discovered. On the basis of performed tests made in our Department we diagnosed connective tissue disease. The patient was treated with glycocorticoids with good clinical response: normalization of hematological changes and disease remission. PMID- 11928572 TI - [Prostaglandin D2: biosynthesis, biological properties and participation in bronchial asthma]. PMID- 11928571 TI - [Ormond's fibrosis, bone osteolysis and stomach intramural metastases in the course f low-differentiated prostatic cancer]. AB - Retroperitoneal fibrosis (Ormond's disease) is rare chronic inflammatory process, that can occur at any age. It is characterised by development of periaortic fibrous mass leading to progressive obstruction of vessels around the abdominal aorta and ureters. In the one third of cases we can find the causes of disease. There are ergotamine abuse, radiation, retroperitoneal surgery or hemorrhage, urine extravasation and response to different cancers. The other cases are idiopathic disease. We report a case of prostate cancer with unique course. The first manifestations of disease were diffuse peritoneal fibrosis and ureteral obstruction leading to bilateral hydronephrosis. Clinical course and histopathology showed idiopathic Ormond's fibrosis. Patient received oral immunosuppressive treatment (prednisolone 1 mg/kg/day + azathioprine 1 mg/kg/day), followed by intravenous methylprednisolone puls (2 g). Treatment also consisted of DJ-stent placement on the left side. On the right side we were unable to overcome the obstruction of ureter. Because of persistent renal failure, thrombocytopenia, DIC and progressive lower back pain we did control MR and CT scan. The CT scans showed multiple osteolytic bone metastases in vertebral column (the sizes of them were between a few millimetres and 1.5 centimetre). Patient died due to renal failure and haemorrhagic diathesis in the course of disseminated cancer of unknown origin. The postmortem examination revealed diffuse peritoneal infiltration surrounding the ureters, intramural ventricular metastases, pulmonary metastases and vertebral metastases. The prostate was only slightly enlarged. Histological and immunohistochemical examinations of prostate showed primary low-differentiated prostate carcinoma (CK/+/, PAP/+/, PSA/+/). Peritoneal, ventricular and bone infiltrations also were metastases from low differentiated carcinoma of prostate origin (CK/+/, PAP/+/, PSA/-/). PMID- 11928573 TI - [Pseudoxanthoma elasticum]. PMID- 11928575 TI - [Psoriatic arthritis]. PMID- 11928574 TI - [Inhibition of bone resorption by bisphosphonates, heterogeneous group of drugs with different intracellular model of action]. PMID- 11928576 TI - [Professor Akihiro Igata awarded with the Medal of the Polish Society of Internal Medicine]. PMID- 11928579 TI - CareCredit: helping you get paid faster. PMID- 11928580 TI - Oral pathology quiz #4. Ectopic geographic tongue. PMID- 11928581 TI - Understanding HIPAA: the Health Insurance Portability and Accountability Act. PMID- 11928578 TI - Coding for routine periodontal treatment: Part 3. PMID- 11928582 TI - [Polish Multicenter Study on Diabetes Epidemiology (PMSDE)--1998-2000]. AB - Diabetes mellitus is a growing health problem at all ages in all countries. The Saint Vincent Declaration signed by the representatives of all European countries -among them by Poland--indicated the main goals to reduce this burden. This may be achieved by elaboration and initiation of the nation-wide programme for early detection and control of diabetes on the population level. The Polish Task Force for Diabetes Management worked out and put into operation in 1998 the project commissioned by the Ministry of Health funded by the Polish State Committee for Research (PBZ-018-11). The urgent need for the implementation of this programme was a result of the increasing number of diabetic patients on the primary health care level, diabetic complications and hospitalisations. The programme was based on the assumption that primary and secondary prevention of NIDDM may eliminate or reduce its risk factors and therefore decrease its prevalence and morbidity and should integrate epidemiology, identification of risk factors, education, intervention models and economics of diabetes. This issue of the Polish Archives of Internal Medicine contains the main results of the programme and reflects an actual situation of diabetes in Poland creating a rational basis for intervention on the population level. Evaluation of the incidence rate of the diabetes type 1 in 10 selected areas (population basis 30% of age group 0-29) revealed values from 8.4 to 14.7/100,000 in the age group 1-14 and from 4.4 to 11.2/100,000 in the age group 15-29. It accounts for the 2-3 fold increase in comparison with the results achieved in 1986 (Z. Szybinski). However an ascertainment of the register in the age group 15-29 dropped down during the last 10 years and additional sources of data must be involved in the survey. Prevalence of diabetes type 2 in 3 selected areas Krakow 10.77% (M. Szurkowska), Lublin 15.6% (J. Lopatynski) and Lodz 15.7% (J. Drzewoski) based on the population random sample over 35 years of age, constitutes over 90% of all patients with diabetes. When standardized to the Polish population the prevalence of diabetes type 2 accounts for 5.37% and reflects a true endemic state. With comparison to a similar survey carried out in 1986 in Wroclaw the prevalence 3.71% of the diabetes type 2 increased 2-3 times. It allowed to calculate an actual number of patients with diabetes type 2--over 2 million and around 50% of them represents "unknown" diabetes. Such high contribution of the unknown non-diagnosed diabetes has been confirmed in the survey carried out in Poznan on the non-randomized professional group of persons (B. Wierusz-Wysocka) in investigation on the first-degree relatives of patients with known diabetes in Warsaw (T. Kasperska-Czyzyk) and in Wroclaw (R. Wasikowa). This raises the question about the criteria of diagnosis for unknown diabetes. Comparative analysis carried out in Krakow revealed that fasting glycemia 7.0 mmol/L allowed to diagnose diabetes with 50% with respect to the 2 hours oral glucose tolerance test (WHO 1985) and shouldn't be use as a tool for early diagnosis of diabetes type 2 (Z. Szybinski, M. Szurkowska) Identification of the risk factors in PMSDE for diabetes type 2 allowed to introduce the term "global risk factors", divided into two groups primary (obesity, age, pregnancy and genetic background) and secondary (hyperinsulinemia, hyperglycemia, dyslipidemia and hypertension). In the multifactor analysis obesity and hyperinsulinemia are the strongest predictors and modifiable risk factors of the development of diabetes type 2 and late complications. Especially hyperinsulinemia as an independent secondary risk factor for hypertension, dyslipidemia--in consequences late diabetic complications and perhaps carcinogenesis may play an important role as a predictor of diabetes type 2 and biochemical marker of effectiveness of non pharmacological and pharmacological approach in the global concept of diabetes type 2 treatment (Z. Szybinski). Analysis of the late complications of diabetes were focussed on visual disability and lower extremity amputation due to diabetic foot development. Analysis of prevalence (PR) of visual disability due to diabetes in Krakow (J. Pantoflinski) and Olsztyn (E. Bandurska-Stankiewicz) revealed similar values, in the both areas around 6% of general visual disability in population. Monitoring of the visual disability in diabetic patients seems to be a good indicator of the quality level of the diabetic care. Analysis of the lower extremity amputation carried out in Krakow district (A. Nazim) revealed that incidence rate of amputation in diabetics was 15 times higher than in non diabetics and in 10.8% of cases amputation was performed in unknown diabetes type 2. The preventive measures and non-pharmacological and pharmacological management approximately in diabetes should be solved with education based on the separate post of a diabetic educator within a diabetic team and on the primary health care level. The articles presenting elaborated educational model in diabetes are published in "Diabetologia Polska" (A. Czyzyk). Intervention model was tested in Krakow in the groups of obese patients with newly diagnosed "unknown" diabetes and was based on the 12 weeks supervised dietician education with standardized physical activity programme. In obese diabetic patients the weight loss less than 10% of the initial body weight can markedly improve biochemical parameters like hyperglycemia dyslipidemia and hyperinsulinemia without pharmacological therapy and 45% of patients has been transferred to the group of Impaired Glucose Tolerance (A. Gilis-Januszewska). Another model of intervention may be applied in menopausal women (J. Nadel, K. Cypryk) treated with hormonal replacement therapy as a complementary factor to education and if necessary to anti-diabetic drugs. Increase number of diabetic patients especially of the "unknown" type 2 has serious economic aspect by increasing a burden of patients, theirs families, society and health care system. Within PMSDE programme elaboration of the model for calculation the direct costs about of diabetes and burden in terms of years of life lost using DALY measure was performed (K. Kissimova-Skarbek). Average diabetes type 1 patients costs 7 times and type 2 over 3 times higher than average health care cost and 95% of total time lost due to disability is caused by diabetes type 2. Therefor primary and secondary prevention of diabetes typ 2 have highest priority among strategic preventive targets. The Review Conference held in Warsaw at 24-25 February 2001 in the presence of WHO Experts formulated the recommendations focused on: 1. Elaboration of high risk strategy for early diagnosis of unknown diabetes type 2 based on the 2-hours OGTT (WHO 1985) 2. Continuation of the epidemiologic study in diabetes 3. Instituting the professional post for diabetes educator on the specialized and primary health care levels. 4. Further research are recommended for evaluation of the role of fasting glycemia and hyperinsulinemia as predictors and risk factors of diabetes type 2, for development of preventive models in diabetes type 2 and for development of the economical models to asses the costs of diabetes (Recommendations). PMID- 11928583 TI - [Polish multicenter study on diabetes type I incidence in he age group 0-14 between 1998 and 1999]. AB - The increase in diabetes type 1 incidence observed in various centers in Poland and the need for a centralized study covering large population have resulted in the construction of a standardized registry of type 1 diabetes in 1998 within the Polish Multicenter Study in Diabetes Epidemiology. The aim of the study was to present the incidence rates of type 1 diabetes in the age group 0-14 in 7 distinct regions of Poland (Krakow, Wroclaw, Warsaw, Bialystok, Poznan, Rzeszow and Olsztyn centers) with over 30% of the Polish population at risk in 1998 and 1999. The data for the standardized registry were obtained prospectively from paediatric hospital wards and diabetes outpatient units. The incidence rates calculated in 1998 showed the highest value of 14.6 and 14.5/100,000 for Olsztyn and Warsaw, and the lowest (8.4/100,000) for Poznan center. In 1999 the highest value of 14.7/100,000 was noted in Krakow and the lowest (9.3/100,000) in Poznan center. The differences in diabetes type 1 incidence rates between age groups 0 4, 5-9 and 10-14 were found to be significant (p < 0.0005) and were also significant when incidence rates were compared between males and females in these age groups in the whole study area in 1998-1999 (p = 0.002 and p = 0.015 respectively). PMID- 11928584 TI - [Polish multicenter study on type I diabetes incidence in the age group 15-29 between 1998 and 1999]. AB - The increase in diabetes type 1 incidence observed in various centers in Poland and the need for a centralized study covering a large population have resulted in the construction of a standardized registry of type 1 diabetes in 1998 within the Polish Multicenter Study in Diabetes Epidemiology. The aim of the study was to evaluate the incidence of type 1 diabetes in the age group 15-29 in 5 distinct regions of Poland (Krakow, Warsaw, Bialystok, Rzeszow and Olsztyn centers) with over 15% of the Polish population at risk in 1998 and 1999. The data for the standardized registry were obtained prospectively from hospital departments and diabetes outpatient units. The incidence rates calculated in 1998 showed the highest value of 11.2/100,000 for Krakow and its region, and the lowest (4.4/100,000) for Bialystok and its region. In 1999 the highest value of 12.3/100,000 was noted in Olsztyn and its region and the lowest (3.4/100,000) in Warsaw. There were significant differences in the incidence rates between the study centers were found. Incidence rates in the whole study area were significantly higher among males as compared with females in 1998 and 1999 (8.9/100,000 vs. 4.9/100,000; p = 0.0001), marked in the age groups 15-19 and 20 24 (p = 0.001 and p = 0.002 respectively). A significant increase in diabetes type 1 incidence (from 4.6/100,000 to 6.9/100,000) was found as compared with results of the "Three Cities Study" (1986-1988). PMID- 11928585 TI - [Prevalence of type II diabetes mellitus in population of Krakow]. AB - The study was carried out within a framework of the Polish Multicenter Study on Diabetes Epidemiology in 1998-2000. The aim of the study was to define the prevalence of type 2 diabetes, especially unknown diabetes, and prevalence of impaired glucose tolerance in a demographically well-defined urban population using the comparable epidemiological methods which were applied in the previous study in Wroclaw in 1985-1986. The study was carried out in 200,000 subjects inhabiting the town quarter. Out of those who were 35 or more 6000 subjects were randomised using a table of random numbers. All randomized subjects received a letter of invitation explaining the sense of study, its objectives and methods. If necessary the invitations were renewed, and then the subjects were contacted by phone. Each responding person received a questionnaire to complete. Then anthropometric and blood pressure measurements were taken. Blood was sampled for plasma glucose, insulin, total cholesterol, HDL cholesterol and triglycerides in the fasting state. Those who declared being non-diabetic and in whom screening test using a glucometer (Glucotrend) revealed fasting glycemia below 8 mmol/l underwent an oral glucose tolerance test (75 g) to determine glycemia and insulinemia at 120 min. Plasma glucose, total cholesterol, HDL cholesterol and triglycerides concentrations were measured with an enzymatic method, whereas insulinemia was defined with the IRMA technique, using ready kits Swierk-Poland. Diabetes mellitus and impaired glucose tolerance were recognised according to the 1985 WHO criteria. Chi square test, Fisher's test and Mann-Whitney test were used for statistical analysis. Statistical analysis was carried out using the statistical package BMDP. During 3 years of the study out of 6000 randomly selected subjects 3060 (1731 women and 1329 men) responded. In the study population 192 patients were with known diabetes, including 150 subjects receiving oral antidiabetic agents or insulin at the time of the study or some with high fasting glycemia not receiving any treatment except a diet. The 42 subjects who prior to the study had not been receiving hypoglycemic agents or in whom fasting glycemia had been below 8 mmol/l underwent an oral glucose tolerance test. Of them diabetes was confirmed in 11 patients, impaired glucose tolerance was observed in 9, and glucose intolerance was excluded in 22 subjects. Thus, in the study group 161 subjects (75 women and 86 men) with a mean age 61.5 +/- 8.95 years had already diabetes. Their mean BMI was 31.5 +/- 4.6 kg/m2 and did not differ significantly between both sexes. Only HDL cholesterol was significantly higher in men (women 1.1 +/- 0.3 vs. men 1.3 +/- 0.3, p < 0.001) in this group. Among those who declared being non-diabetic 160 subjects (77 men and 83 women), mean age 58.0 +/- 9.7 years and mean BMI 31.4 +/- 4.9 kg/m2 had diabetes identified according to the 1985 WHO criteria. Fasting insulinemia was 16.6 +/- 12.0 uj/ml in this group. At 120 min OGTT insulinemia in women was higher than in men (152.6 +/- 90.5 vs. 112.0 +/- 83.4, p < 0.01). In the whole study population diabetes was found in 321 subjects, including 161 with known and 160 with newly diagnosed diabetes. Based upon these data a standardized prevalence rate due to type 2 diabetes was calculated being 5.37% for the whole population (2.82% for known and 2.55% for unknown diabetes, respectively). When only part of the population over 35 years of age was taken into consideration, the rate was 10.77% (5.66% for known and 5.11% for unknown diabetes). When only fasting glycemia according to ADA recommendation was analysed, diabetes was recognised in 160 subjects (107 men and 53 women). In 78 subjects (49 men and 29 women) diabetes was diagnosed according to the WHO and ADA criteria. When oral glucose tolerance test and glycemia at 120 min exceeding 11.1 mmo/l is considered a gold standard for the diagnosis of diabetes, the diagnostic accuracy of the ADA criteria is 48.7%. In the study population 449 (14.55%) subjects (201 men and 248 women), mean age 56.6 +/- 9.6 years and mean BMI 29.7 +/- 4.6 (men 29.0 +/- 3.7 vs. women 30.2 +/- 5.2, p < 0.01) had impaired glucose tolerance. In our study population there were 572 subjects (329 men and 243 women) with impaired fasting glucose. Of them 359 subjects (212 men and 147 women) had normal glucose tolerance in OGTT, 161 (99 men and 62 women) had impaired glucose tolerance, and 52 (18 men and 62 women) type 2 diabetes. Thus, of the 572 subjects 9% (5.4% of men and 13% of women) had diabetes type 2, and 28% (30% of men and 25% of women) had impaired glucose tolerance. As the frequency of impaired glucose tolerance in this subgroup is higher than in the whole study population it seems justified to identify a group of subjects with increased fasting glycemia and to administer OGTT. CONCLUSIONS: 1. A significant rise in the prevalence of type 2 diabetes was observed between 1986 and 2000 (from 3.7% to 10.77%). 2. Prevalence of unknown diabetes increased considerably (reaching 5.11%). 3. The similar rise in the prevalence of impaired glucose tolerance was observed between 1986 and 2000 (from 2.9% to 14.5%) 4. Early detection of type 2 diabetes should be based upon oral glucose tolerance test according to the WHO. PMID- 11928586 TI - [The prevalence of type II diabetes mellitus in rural urban population over 35 years of age in Lublin region (Eastern Poland)]. AB - The prevalence of type 2 diabetes mellitus (DM 2) has increased dramatically in the last decade. Data relating to the number of undetected cases of diabetes are underestimated. The aim of the study was to evaluate the prevalence of DM 2, obesity, hypertension, and lipid disturbances in a representative group of urban and rural population in the Lublin region (Eastern Poland). The study was performed in 1998-2001. A two-layer draw was applied: two groups of 3000 people were drawn from the population of Lublin town and from the rural areas each comprising 100,000 inhabitants. In all subjects physical examination was performed and body weight, height, and blood pressure measurements were obtained. Blood samples were taken from the basilic vein to estimate: blood glucose, lipids and insulin concentration. Venous blood glucose concentration was measured using a Glucotrend glucometer. Oral glucose tolerance test (OGTT) after a 75 g-glucose load was performed in subjects without previously diagnosed diabetes mellitus and when the fasting blood glucose was < 8.0 mmol/l (144 mg/l). The LDL-cholesterol level was calculated according to Friedewald formula. DM 2 was identified according to the WHO criteria from 1985. Obesity and hypertension were diagnosed according to the new WHO criteria (Body Mass Index > or = 30 kg/m2, blood pressure > or = 140/90 mm Hg). 3782 subjects: 1809 in the rural area and 1973 in Lublin town were examined. The response rate among rural and urban population was 60.3% and 65.8% respectively. The prevalence of DM 2 was assessed in 17.6% of rural and in 14.1% of urban population. 75% of diabetics in the rural areas and 56% in the town were the newly diagnosed cases. We found impaired glucose tolerance in 30.3% of rural and in 21.6% of urban population, BMI > or = 30 kg/m2 in 30.8% and 30.1%, hypertension in 69.4% (29.2% newly diagnosed) and 68.6% (27.7% newly diagnosed), hypercholesterolaemia (total cholesterol > or = 5.2 mmol/l (200 mg/dl)) in 66.4% and 60%, hyper-LDL-cholesterolaemia (> or = 3.5 mmol/l (135 mg/dl)) in 57.3% and 52.6%, hypo-HDL--cholesterolaemia in 21.7% and 31.4%, hypertriglyceridemia (> or = 2.3 mmol/l (200 mg/dl)) in 15.1% and 22% respectively. This finding indicates the urgent need for introducing a national program for early diagnosis and prevention of DM 2 and concomitant metabolic disturbances. PMID- 11928587 TI - [Type II diabetes mellitus and selected metabolic disorders in urban population aged over 35 years]. AB - Recent epidemiologic studies show increasing "epidemic" of diabetes mellitus throughout the world. Reliable data on diabetes prevalence in Poland are scarce. Therefore Polish Ministry of Health initiated a programme aiming at gathering substantial amount of epidemiologic data on the prevalence of diabetes and other metabolic disorders. The aim of the present study was to evaluate the prevalence of diabetes, obesity and lipid disorders in a well-defined urban population aged 35 and over. The study subjects were 6000 randomly chosen inhabitants of the Central District of Lodz. All were invitated to participate in the study by mail. Every participant underwent full medical examination, with body mass index (BMI) calculation, and blood pressure as well as waist-to-hip ratio measurements. Serum total, LDL and HDL cholesterol and triglycerides were assessed. In non-diabetes subjects oral glucose tolerance test (75 g) (OGTT) according to WHO protocol was performed unless their fasting capillary blood glucose exceeded 8 mmol/l. In selected subjects serum samples were stored for future insulin and C-peptide assays. 2018 persons took part in the study (response rate 33.6%), including 1217 (60.3%) women and 801 men (39.7%), mean age 58.2 years. 179 (8.9%) persons claimed to have been diagnosed with diabetes previously (8.9%). OGTT was performed in 1574 subjects. Impaired glucose tolerance (IGT) was found in 342 (17.0%), and diabetes in 138 (6.8%) subjects. Total diabetes prevalence reached therefore 15.7%. Excessive body weight (BMI > or = 25 kg/m2) was noted in 806 (39.9%), and obesity (BMI > or = 30 kg/m2) in 626 (31.0%) persons. Total cholesterol > 5.2 mmol/l was observed in 1170 (58.0%), LDL-cholesterol > 3.5 mmol/l in 734 (36.4%), cholesterol HDL < 0.9 mmol/l in 953 (47.2%), and triglycerides > 1.7 mmol/l in 1392 osob (69.0%) subjects. In conclusion, high prevalence of known and unknown diabetes together with other metabolic disorders is strikingly high in adult urban population, which in all may require effective implementation of specific nationwide prevention programmes. PMID- 11928588 TI - [Insulinemia--a marker of early diagnosis and control of efficacy of treatment of type II diabetes]. AB - Metabolic Syndrome X defined by Reaven is caused by peripheral insuline receptor resistance, leads to hyperinsulinemia regarded as a cause of secondary dyslipidemia, hypertension, hemostatic disturbances, atherosclerosis and insulin as a growth factor takes part in carcinogenesis. Depending on a contribution of the primary risk factors of type 2 Diabetes Mellitus (2-DM) mainly genetic factors and obesity--an independent cause of insulin receptor resistance--glucose intolerance and 2-DM may overlap the Syndrome X. The aims of these studies were to determine in cross-sectional investigation a plasma insulin concentration in subjects aged over 35 years and to assess the clinical usefulness of insulinemia in early diagnosis of diabetes type 2. Investigations were carried out in Krakow town's district with 200,000 inhabitants, out of those 3060 randomly selected subjects (1720 females and 1340 males aged over 35 years) took part in the Polish Multicenter Study on Diabetes Epidemiology (PMSDE) with protocol and methods previously presented. Glucose concentration was determine by enzymatic method, insuline in plasma by IRMA method using ready kits produced by the Swierk-Poland. Logistic multiple regression model was used to estimate the effect of risk factors on the development of glucose intolerance, Chi square test, Fisher test and Mann-Whitney test were used for statistical analysis by means of statistical package BMPD. Fasting insulinemia in persons with normal glucose tolerance and body weight (BMI < 25 and glycemia < 6.1 mmol/l) in subpopulation aged over 35 years was 5.73 (SD = 3.99) in men and 7.05 (SD = 4.67) microU/ml in women. These values were positively correlated with BMI and at the range 25-30 and > 30 increased by 50 and 100% responsively and in 2-nd h in OGTT by five-times. In the persons with glucose intolerance and new-diagnosed 2-DM insulinemia increased 2-3 fold depending on BMI, and gender. In the subgroup with 2-DM and BMI > 30, insulinemia in 2 h-OGTT treated values 152 (SD = 90) in women and 112 (SD = 83.4) microU/ml in men. Obesity and insulinemia in 2 h-OGTT in multiple analysis have been identified as a strong predictors and risk factors of impaired glucose intolerance (IGT) 2-DM fasting insulinemia may be useful as an indicator of the peripheric insulin receptor resistance. The results lead to the conclusions that determination of the plasma insulin concentration may be useful in early diagnosis of IGT and diabetes type 2, and should be monitored in the course of non-pharmacological and pharmacological treatment 2-DM. One of the main goals in the course of treatment of obesity and early phases of the 2-DM should be normalization or at least reduction of hyperinsulinemia. Insulinemia may be regarded also as an important criterion for selection of the oral antidiabetic drugs. PMID- 11928589 TI - [Strategies for early detection of the risk of type II (non-insulin-dependent) diabetes in 1st degree relatives of patients with this disease]. AB - The present study included two groups of subjects: I. the adult offspring of parents with conjugal type 2 diabetes (n = 77; age range 18-59 yrs and mean age 38 +/- 0.8 yrs; BMI range 18.9-40.3 kg/m2 and mean value 26.6 +/- 0.6 kg/m2); and II. the adult offspring having one parent with type 2 diabetes: either father (n = 83-53%) or mother (n = 74-47%). The age range of the latter group was 21-64 yrs, mean age 41 +/- 0.8 yrs; BMI range was 17.6-46.4 kg/m2, and mean value 26.8 +/- 0.4 kg/m2. The normal glucose tolerance of the "healthy" parent was verified with the OGGT evaluated by the WHO and ADA criteria. In all offspring the same test (75 g) was performed, and glucose in venous blood and insulin (IRI) in serum were determined on fasting and at 30, 60 and 120 min of the test. In fasting state the levels of serum lipids (triglycerides, total and LDL and HDL cholesterol and apolipoprotein AI and B) were also measured. In the group I unknown diabetes mellitus was discovered in 4 cases (4%): in 3 according to the WHO/ADA criteria and in one case evaluated by the ADA criteria), in 19 subjects (25%) IGT was found in 16 isolated and in 3 associated with isolated fasting glycaemia (IFG), and only in one case (1%) the isolated IFG was ascertained. In the group II diabetes was discovered according to the ADA criteria in 4 persons (2.5%), IGT in 29 subjects (18.5%), of whom 8 had also IFG. In this subgroup 16 subjects had diabetic father and 13 diabetic mother. The isolated IFG had 7 offspring (4.4%), of whom 2 had diabetic father and 5 diabetic mother. Apart from glycaemia, other parameters didn't disclose difference between the offspring of diabetic father and diabetic mother. However, considering these parameters together for the whole group II, it was found that the offspring with IGT, isolated and associated with IFG, differed from the remaining ones with significantly higher BMI, higher systolic blood pressure, higher 2-h serum IRI, and higher levels of serum triglycerides, total cholesterol and ApoB. CONCLUSION: Measurement of isolated fasting glycaemia and its interpretation by the ADA criteria is inadequate in studies aiming at early detection of glucose intolerance in subjects with familial increased risk of type 2 diabetes and should include also the determination of the 2-h glycemia of the OGTT evaluated according to the WHO criteria. On the other hand, the determination of fasting glycaemia and its evaluation by the ADA criteria is a valuable in diabetes screening, as its elevated level may identify other metabolic disorders associated with diabetes, and unfavorable risk of cardiovascular complications of this disease. PMID- 11928591 TI - [Appearance of undiagnosed diabetes mellitus in the population of professionally active people in the urban areas]. AB - Diabetes remains a great social and clinical problem. Therefore, there is a need to focus our efforts on prevention of the disease, especially of type 2 diabetes. Type 2 diabetes is characterized by accelerated development of atherosclerotic changes (macroangiopathy). Hyperglycaemia, hypertension, hyperlipidaemia and hyperfibrinogenaemia also play an important role in the development of macroangiopathy. Hyperinsulinemia, which accompanies the visceral type of obesity, is characteristic of type 2 diabetes. Considering all the above mentioned findings, prevention of type 2 diabetes should be based on the population level, concentrating especially on the groups with increased risk of obesity and/or diabetes (early primary prevention). However, in the present conditions, it seems that screening studies can be conducted only in the groups with high risk of type 2 diabetes (late primary prevention). They allow for relatively early detection of disturbances in carbohydrate metabolism. The aim of the study was to assess the prevalence of undiagnosed diabetes in the population of professionally active inhabitants in Pleszew. 2700 subjects, aged 35-65 years, entered the study. All patients claimed to be healthy. In the first phase of the study, the fasting capillary glycaemia was tested. Fasting blood glucose or oral glucose tolerance test was performed in all cases which fasting capillary glucose was higher then 5.5 mmol/l (100 mg/dl). The screening study revealed 91 cases with glycaemia higher than 6.8 mmol/l (3.4%). 387 subjects (14.3%) with glycaemia ranging from 5.5 to 6.8 mmol/l were qualified to perform the oral glucose tolerance test. Out of this group 138 persons did not come to the laboratory. Thus, the test was conducted in 249 causes (64.3%). The results obtained excluded another 197 subjects as no disturbances in the glucose metabolism were found. Based on the results of the oral glucose tolerance test 39 patients were diagnosed to have an impaired glucose tolerance (2 h glycaemia from 7.8 to 11.1 mmol/l) and in 13 cases diabetes was diagnosed (2 h glycaemia above 11.1 mmol/l). In conclusion, the screening study performed in professionally active adults aged > 35 years, who claimed to be healthy, clinically latent diabetes or impaired glucose tolerance was found in 5.3% cases. 92.8% patients with IGT or diabetes were obese or overweight (BMI > 25 kg/m2) and 32.4% had hypertension (RR > 140/90 mm Hg). In 64% of subjects the serum cholesterol concentration was higher than 5.2 mmol/l and in 18% subjects HDL cholesterol concentration was lower than 1.0 mmol/l and LDL cholesterol higher than 3.5 mmol/l. Elevated triglycerides concentration > 2.0 mmol/l was observed in 30%. In the group with newly diagnosed diabetes, mean age was 55.0 +/- 9.2 years. 27.9% had positive family history of diabetes, 26.5% were smokers, 44.1% were found to have disturbed lower limbs circulation and 30.9% had abnormal feeling of vibration, 7.8% patients with diabetes had symptoms of diabetic retinopathy and 20.1% had microalbuminuria. Body mass index (BMI) in newly diagnosed diabetic patients was 31.6 +/- 5.3 kg/m2 and waist to hip ratio (WHR) was 0.94 +/- 0.41 and indicated the visceral type of obesity. Mean fasting glycaemia was equal 7.26 +/- 1.93 mmol/l and mean HbA1c value was 6.2 +/- 0.7%. It exceeded the laboratory normal value in 17.6% of cases. In 91 patients with fasting glycaemia higher then 15.5 mmol/l insulinaemia was also assessed; its level was elevated in 10 patients. The project of study was prepared in 1996. However, in 1999 the new criteria for diagnosis and treatment of type 2 diabetes were established. The results of the performed study indicate that screening towards diabetes should be performed in subjects aged > 35 years with overweight or obesity and at least one additional risk factor of arteriosclerosis. PMID- 11928590 TI - [Risk of incidence of diabetes mellitus in relatives of patients with diabetes type I in retrospective questionnaire study]. AB - It is commonly known that there is a higher risk of diabetes type 1 in relatives of patients with diabetes type 1. According to some reports in families of these patients the incidence of diabetes type 2 is also higher. The aim of our study was the evaluation of incidence of diabetes type 1 and 2 in 1st and 2nd degree relatives of patients with diabetes type 1. Our study was conducted in the years 1993-2000 in the Department of Endocrinology for Children and Adolescents in Wroclaw and in the Department of Endocrinology of Jagiellonian University in Cracow among relatives of all the patients in the age of 0-19 years with newly diagnosed diabetes type 1. Special prepared questionnaires were used in which patients were asked about: number of relatives of 1st and 2nd degree, age, sex and diagnosis of diabetes in the relatives. Data from families of 332 patients were obtained. They concerned 4080 relatives. Diabetes occurred in 121 relatives (2.96%). 20 of them were 1st degree relatives and 101--2nd degree relatives. In 31 relatives (0.76%) diabetes type 1 was diagnosed and in 88 relatives (2.16%) diabetes type 2 was diagnosed. Except for one individual (patient's mother) diabetes type 2 occurred in 2nd degree relatives. Diabetes type 1 was diagnosed in 16 1st degree relatives and 15 2nd degree relatives. They were: 1st degree relatives: 9 fathers, 3 brothers, 2 mothers and 2 sisters. 2nd degree relatives: in 8 cases siblings of patient's parents, in 7 grandparents. In families of diabetic children and adolescents relatives with type 1 and type 2 diabetes were observed, with a dominance of relatives with diabetes type 2 in 2nd degree relatives. PMID- 11928592 TI - [Studies on the incidence and clinical significance of the metabolic syndrome in postmenopausal women in Lodz region]. AB - In menopausal period increased prevalence of lipid metabolism disorders, atherosclerosis, carbohydrate metabolism disorders, cardiovascular diseases is observed. AIM OF STUDY: Evaluation of prevalence of obesity, overweight, hypertension, carbohydrate and lipid metabolism disorders in population of women aged 45-65, treated in Menopause Diagnostic and Treatment Center Polish Mother's Memorial Hospital--Research Institute in Lodz. 2081 women, aged 45-65 were studied. In all of them diseases history, body mass index (BMI), blood pressure (twice), total cholesterol, triglycerides, fasting blood glucose levels were obtained. The highest percentage of obese women was noted in small villages (39%), whereas 21% of women from cities have BMI > or = 30 kg/m2. On the other side only 18.4% women living in cities have normal body weight, although in small villages there are 34.6% women from this group. Overweighted women percentage is similar among women living in small villages and cities. Hypertension prevalence did not depend on place of living, but obese women have hypertension more frequently. In study group diabetes mellitus was diagnosed in 4.8% of women and impaired glucose tolerance in 7%. Almost 2/3 of diabetes mellitus cases was first time diagnosed. Fasting blood glucose (FBG) level > or = 125 mg/dl was noted in 62 women (3%), values 111-124 mg/dl in 124 (6%), and in other 1895 women (91%) FBG was < or = 110 mg/dl. FBG level over 124 mg/dl and diabetes mellitus was more frequent in obese women. Women with hypercholesterolemia mostly (63.2%) have BMI > 25 kg/m2. Our study revealed that hypercholesterolemia prevalence did not differ with no statistical significance between women living in cities, towns and small villages. Elevated triglycerides level over 2.26 mmol/l was noted in 14.5% studied women, whereas there was no correlation between triglycerides level and body weight and place of living. Main health problem of women in perimenopausal period is obesity and lipid metabolism disorders detected in significant number of studied patients. Most cases of undiagnosed diabetes mellitus indicate that complex evaluation of health status of women in perimenopausal period is obligatory procedure. PMID- 11928593 TI - [Incidence of lower extremity amputations in diabetics]. AB - The aim of this study was to identify the incidence of lower extremity amputations (LEAs) in diabetics in Poland as background data for comparisons over time and with other communities. This was a cross-sectional study of the incidence of any non-traumatic and non-neoplastic LEAs in the population of Krakow region (1,239,703 inhabitants in its borders before 1.1.1999). It was calculated that for this population size it was sufficient to collect data for one year. The amputees had to be inhabitants of the Krakow region between 1.1.1996 and 31.12.1996. Data was collected from two sources: surgical wards and limb fitting centres. 290 non-traumatic and non-neoplastic LEAs were identified: 283 in the first source (hospital), of which 51 were found in the second (limb fitting centre) while 7 amputations were identified in the second source solely. 72.4% of LEAs were performed in males and this proportion tended to decrease with age, especially in diabetics. Mean age of the amputees was 64.7 years. Diabetics were significantly older (68.2 vs 61.5 years, p < 0.001). The number of amputations was growing with age reaching peak values between 65-74 years in diabetics and 55-64 in non-diabetics. 47.9% of LEAs were performed in diabetics. In 10.8% of cases diabetes was previously unknown. 88.7% of amputations were primary. The following amputation levels were identified: toe--15.5%, metatarsus- 6.6%, ankle 1.0%, crus--20.0%, thigh--56.9%. In diabetics 21.6% of amputations were minor (metatarsus and below) comparing to 9.9% in non-diabetics. Incidence rates, calculated per 100,000/year and corrected using capture-recapture method, were as follows (95% confidence intervals in parentheses): 25.9 (10.2-41.6) in the whole population, 186.7 (125.3-248.1) in diabetics, 165.3 (110.5-220.0) primary in diabetics, and 14.4 (3.5-25.4) in non-diabetics. Comparing to data from the literature these were comparatively good results for diabetics of caucasian race. Like in other papers incidence rate in diabetics was almost 15 times higher than in non-diabetics, reaching 24 in females, which suggested that gender protection was reduced by diabetes. In an attempt to lower the rate of amputations several actions should be considered. They may include more active case finding, particularly by GPs, and more effective screening for a high risk diabetic foot. PMID- 11928594 TI - [Visual disability incidence among diabetes patients in Olsztyn province in the years 1990-1999]. AB - The aim of the study was to estimate the incidence of visual disability. Long term studies on visual disability have been conducted in Olsztyn Province since 1989. A standardized register has comprised 292 patients with diabetes type 1 and 2 whose visual disability resulted from diabetes. Visual disability incidence was expressed by means of a standard rate describing the number of newly diagnosed cases of visual disability per 100,000 population per year. Diabetes related visual disability incidence rate ranged from 2.52/100,000 population in 1990 to 3.08/100,000 population in 1999. For diabetes type 1, visual disability incidence rate was 1.20/100,000 population in 1990 and 1.03/100,000 population in 1999. For diabetes type 2, the rate was 1.33/100,000 population in 1990, increased to 4.66/100,000 population in 1995, and decreased to 2.05/100,000 in 1999. The rate was twice as high among women as among men. CONCLUSIONS: 1. The incidence of visual disability in 1990-1999 was 2.52 to 3.08/100,000 population on average. For type 2 diabetes it was significantly higher than for type 1 diabetes. 2. The monitoring of visual disability is a good indicator of diabetological care. PMID- 11928595 TI - [Visual disability due to diabetes in Cracow voivodeship]. AB - Visual impairment and blindness are major complications of diabetes and are regarded as the most serious disability by most of patients. The purpose of the study was to determine the incidence and prevalence due to diabetes-induced visual disability in 1991-1999 in the former Cracow voivodeship with population of 1,245,047 inhabitants. A register of visual disability was established using independent sources of information. On the end of 1999 the register had included 122 diabetics with visual disability--66 women (54.1%) and 56 men (45.9%). The patients with type 2 diabetes predominated--92 subjects (75.4%). The remaining 30 patients (24.6%) had type 1 diabetes. Grade 1 visual disability was diagnosed in 82 subjects (67.2%) whereas grade 2 visual disability in 40 patients (32.8%). Grade 1 visual disability was defined as visual acuity in a better eye < 0.05 and visual field narrowing < 20%. The criteria of grade 2 were visual acuity after correction in a better eye 0.05-0.1 and visual field narrowing 20-30%. According to the register of the Cracow Branch of the Polish Association of the Blind diabetics made up 6.2% of all visual disability cases. In 1991-1999 the incidence of visual disability due to diabetes did not increase significantly. The mean incidence rate was 0.9/100,000 population. However, there was a significant increasing trend in prevalence with mean annual increase of 0.24/100,000 population (95% CI 0.17-0.31/100,000). PMID- 11928596 TI - [The efficacy of non-pharmacological intervention in obese patients with newly diagnosed diabetes mellitus type II]. AB - In Poland like in other countries we observe an increasing number of diabetes mellitus cases with about half of the patients in whom the disease remains undiagnosed. Therefore it seems necessary to improve early diagnoses and prevention of the disease. The aim of the study was to assess the efficiency of a 3-month non-pharmacological intervention based on diet and increased physical activity in patients with newly diagnosed diabetes type 2 (diagnosis based on oral glucose tolerance test (OGTT), WHO 1999). We investigated 37 newly diagnosed diabetic type 2 patients, 16 men (aged 52.4 +/- 5.4) and 21 women (aged 51.0 +/- 5.7). Anthropometric and biochemical measurements were performed before and after intervention. Two-kilometres Walking Test with an intermediate estimation of VO2max and fitness index (FI) was performed before and after intervention. Total abdominal fat volume (measured from diaphragm to pubis): visceral fat volume (VFV) and subcutaneous fat volume (SFV) (mm3) were assessed according to the standard protocol of NMR abdominal examination. Patients completed 12 weeks of supervised intervention focused on weight reduction, increase of physical activity, changes of nutritional habits. Students t-test, Mann-Whitney test and Spearman's correlation were used for statistical analysis. In women the average weight reduction was 4.7 kg (5.8% of initial body weight), whereas in men 5.9 kg (5.9% of initial body weight). In women VO2 max increased from 23.7 +/- 6.4 to 24.9 +/- 4.8 (ns), and fitness index increased from 78.7 +/- 11.7 to 83 +/- 14.7 (ns). In men VO2 max increased from 22.5 +/- 6.7 to 26.6 +/- 8.6 (ns) and fitness index increased from 55.1 +/- 12.5 to 64.8 +/- 13.7 (p < 0.05). In women the level of fasting glycaemia decreased from 6.47 +/- 1.2 to 4.84 mmol/l +/- 0.6 (p < 0.01) and the level of glycaemia at 120 minutes of OGTT decreased from 13.2 +/- 2.5 to 6.76 +/- 2.7 mmol/l (p < 0.01). The decrease of plasma glucose was accompanied by the decrease of fasting insulin from 19.2 +/- 15.5 to 8.53 +/- 93.2 uj/ml (p < 0.01) and in 120 minutes of OGTT from 148.8 +/- 86.2 to 58.4 +/- 41.0 uj/ml (p < 0.01). In men the level of fasting glycaemia decreased from 8.63 +/- 2.0 to 7.07 mmol/l +/- 2.4 (p < 0.05) and the level of glycaemia at 120 minutes of OGTT decreased from 15.76 +/- 3.2 do 9.3 +/- 5.7 mmol/l (p < 0.01). The decrease of plasma glucose was accompanied by the decrease of fasting insulin from 21.99 +/- 12.6 to 10.1 (3.8 uj/ml (p < 0.05) and at 120 minutes of DGTT from 81.5 +/- 52.7 do 41.6 +/- 21.0 uj/ml (p < 0.05). After the intervention 45% of the patients (57% of women and 31% of men) were non-diabetic (correct OGTT). In men visceral fat volume (VFV) was greater than in women (7642.6 +/- 1774.6 and 4789.9 +/- 1242.0 mm3 respectively (p < 0.01). Subcutaneous fat volume (SFV) was smaller in men than in women (7116.5 +/- 2048.5), in men and 10533.9 +/- 3478.3 respectively (p < 0.01). In women and men a strong (p < 0.01) correlation between waist circumference and visceral fat volume (VFV) (r = 0.573 (p < 0.01) and r = 0.833 (p < 0.01) respectively) and subcutaneous fat volume (SFV) (r = 0.900 (p < 0.01) and r = 0.790 (p < 0.01) respectively) was found. The results of the study confirm that in newly diagnosed diabetic type 2 patients body weight reduction and increased physical activity result in the improvement of biochemical indices. In about one half of patients the early phase of the disease might be reversible due to weight reduction and increased physical activity. The non-pharmacological intervention should be the first intervention undertaken in newly diagnosed diabetic type 2 patients. PMID- 11928597 TI - [Evaluation of selected X syndrome factors during hormonal replacement therapy]. AB - Estrogen deficiency after menopause leads to characteristics changes in the hormonal profile, which may influence lipid carbohydrate and calcium-phosphate metabolism and some elements of homeostasis. AIM OF STUDY: To evaluate the influence of hormone replacement therapy with 2 mg estradiol valerate and 0,15 mg levonorgestrel on carbohydrate and lipid metabolism in women after menopause during 12 months of follow-up. We examined 101 women, mean age 52.9 +/- 4.6 years (range from 44 to 65). HRT was applied in 67 women whereas 34 women were without treatment. All of them had no carbohydrate disturbances. All women underwent clinical examination, and body mass index (BMI), fasting blood glucose, insulin, total cholesterol and triglycerides levels were obtained. At 1 and 2 hours after 75 g glucose challenge (75OGTT) glucose and insulin levels were obtained. During hormone replacement therapy all women noted release or significant decrease of climacteric symptoms. Total cholesterol level was decreased, whereas triglycerides did not change. After 12 months of treatment there was also a significant decrease of all factors relating to carbohydrate metabolism--fasting glucose and insulin, insulin/glucose ratio and area under glucose and insulin curves. In our study--after 6 months of follow-up fasting insulin level, area under glucose and insulin curves were increased whereas fasting blood glucose level remained unchanged. Among women without HRT there were no significant changes in selected lipid parameters and BMI. Triglyceride levels decreased (albeit insignificantly) but, total cholesterol levels did not change. After 12 months glucose level did not change, although other carbohydrate parameters were increased. 1) In the study group there was a statistically significant decrease in total cholesterol levels whereas triglycerides remained unchanged. 2) HRT we significantly decreased of insulin resistance and fasting blood glucose levels as compared with non-group HRT. 3) The present results indicate HRT-induced improvement of lipid and carbohydrate metabolism. 4) Long-term HRT is necessary to improve carbohydrate metabolism. PMID- 11928598 TI - [Evaluation of the burden of diabetes in Poland]. AB - Burden of diabetes in terms of economic costs and life years lost due to premature deaths and disability in Poland is analyzed. This study calculates direct costs of type 1 and type 2 diabetes in Poland in 1998 and burden of diabetes in terms of years of life lost using Disability Adjusted Life Years (DALYs) measure within the Polish Multicenter Study of Diabetes Epidemiology (1998-1999). There is a consequent need to evaluate the burden of diabetes for the society and to develop affordable and cost-effective preventing strategies. The burden of diabetes is examined in terms of resources used by diabetic patients and time lost due to premature deaths and disability caused by diabetes. The profile of "a standard patient" (type 1 and type 2 diabetes) resource utilization is created using patient survey in Krakow. This includes main elements of cost associated with prevention, diagnosis and treatment: ambulatory care (visits); hospital care (bed/days and dialysis sessions); pharmaceuticals (goods consumed) and diagnosis (tests). This study calculates direct costs to the health sector of type 1 and type 2 diabetes in Poland 1998. Burden of diabetes in Poland in terms of time lost in 1998 is expressed in Disability Adjusted Life Years (DALYs) unit of measurement. DALY is a combination of two dimensions: YLL- number of years lost due to premature mortality; YLD--loss of healthy years due to disability caused by diabetes (with and without complications). The incidence approach is applied for the YLD caused by diabetes type 1 calculations by gender and age groups (0-29 years). Incidence rates are obtained from the prospective data collection [1, 2]. Other data as average age of onset, average duration of the disease (with or without complications), severity (age specific disability weight for treated or untreated forms of diabetes--with or without complications) are obtained from the GBD study for the Formerly Socialist Economies of Europe [9]. Discounting and age weighting procedure is applied. The prevalence approach is applied for YLD caused by diabetes type 2 calculations for treated and untreated forms of diabetes (with and without complications) by gender and age groups (35 years and more). Prevalence data are obtained from the Polish Multicenter Study on Diabetes Epidemiology. Age specific disability weights for treated or untreated forms of diabetes (with or without complication) are obtained from the GBD study for the Formerly Socialist Economies. Discounting procedure is not applied (duration of the disease is assumed 1 year). Years of Life Lost are calculated using Polish mortality data and life expectancy at the time of death in 1998. Cost of diabetes study is particularly useful in indicating the magnitude of the costs involved, which tend to be much higher than perceived by the general public. In 1998 the average diabetes type 1 patient's costs were 6.4 times and diabetes type 2 patient's costs 3 times higher than average public direct health care costs. The total costs of diabetes in Poland 1998 accounted for 9.3% of total public health care expenditures. The cost of diabetic patient's estimation indicates the potential benefits of effective medical interventions. Not only mortality rates should be taken into consideration in the creation of health policy and financial planning. Disability of the population is also an important factor, particularly in diseases which do not lead to fatalities. In 1998 112,584 DALYs (46% for males and 54% for females) were lost in Poland due to premature deaths and disability caused by diabetes. 72% of the total was due to disability. Secondary prevention is very important especially for diabetes type 2 patients. 95% of total time lost due to disability is caused by diabetes type 2. National burden of disease evaluation is helpful to develop a justifiable basis for setting priorities in purchasing and investing at central and local levels especially in prevention. PMID- 11928600 TI - [Effect of slow repetitive transcranial magnetic stimulation on depression in patients with Parkinson 's disease]. AB - Parkinson's disease (PD), that has usually been associated with movement disorders, is also associated with depression in about 40% of patients [9]. Transcranial magnetic stimulation (TMS) is a new non-invasive technique for direct stimulation of the cerebral cortical neurons [1]. Several open studies have shown that repetitive TMS (rTMS) at both rapid (rapid rTMSi: > 1 Hz) and low frequencies (slow rTMSi: < 1 Hz) may have antidepressant action [2-6]. The study included 8 patients diagnosed as PD fulfilling the DSM-IV criteria for major depression (5 patients) and dysthymia (3 patients). Magnetic stimulator, 200 Mag Stim, total output 2 T and a circular coil of 90 mm, were used. For ten consecutive days, between noon and 1 p.m. the patients were stimulated with apprx. 80% of the output (1.6 T) at 0.5 Hz. The daily treatment implied stimulation of both sides of the head (first the right, then the left) at four sites (prefrontal, frontal, parietal and occipital regions) with 5 stimulations each site (20 stimulations per hemisphere). Before the beginning of the study, 2 3 hours after the last stimulation (day 10), 7 and 14 days after completion of the treatment, the patients were subjected to scoring on the Hamilton Depression Rating Scale [11] and Unified Parkinson's Disease Rating Scale (UPDRS) [12]. The HDRS values before initiation of rTMS were 19.2 +/- 3.1, with significant fall (p < 0.01) after 10 days of stimulation (14.9 +/- 3.2), 17 days (12.2 +/- 2.7) and 24 days (13.6 +/- 5.3) after the beginning of the study, suggesting that the antidepressive effect persisted even two weeks after discontinuation of stimulation. The UPDRS values were monitored concomitantly. The values on this scale failed to alter significantly. In conclusion, rTMS is a relatively safe and painless method associated with antidepressant action in PD patients. Treatment of depression in PD is of great importance, but the choice of medication is accompanied with numerous limitations [20]. Antidepressant action of rTMS and its maintenance for two weeks after discontinuation of stimulation enables usage of this method in PD in phases of exacerbation of depressive symptoms at least over the period required to reach the full effect of selected medication. PMID- 11928599 TI - [The return of anthrax: "the evil spirit is out of the bottle"]. PMID- 11928601 TI - [Use of the partogram in Serbia and its characteristics]. AB - Graphic presentation of parameters during labor is done in Serbia since seventies [1]. Today partogram use is obligatory [2]. Choice of parameters and way of their presentation depend on institution in which they are being used [1]. However, the rules of graphic presentation should be obeyed when constructing cervigrams (graphs representing dilatation vs time). Furthermore, Cartmill and associates [3] found that the slope of the line which represents dilatation directly influences decisions of obstetricians towards interventions during labor. Steeper line reduces the number of decisions for intervention. Importance of these findings is even greater when one has in mind the known inconsistencies in obstetric decision-making [4]. METHODS: From larger maternity hospitals (more than 1,000 labors per year) [5] information concerning use of partograms and partograms themselves were requested. Cervigrams were analyzed in order to evaluate labeling of axes, unambiguity in labeling divisions on scales and the slope of the dilatation line for dilatation of 1 cm/h. Evaluation of parameters which influence the precision of entering data into graphs such as the smallest units on scales and dimensions of 1 cm x 1 h fields in mm, was done. RESULTS: The number of labors in central Serbia and Vojvodina is approximately 85,000 per year [6]. Of 26 maternity hospitals with more than 1,000 labors per year 13 use partograms during labor. Partogram forms were received from 11 maternity hospitals. At least 48% of all births in central Serbia and Vojvodina take place in maternity hospitals which use partograms. Larger maternity hospitals tend to use partograms in greater proportion than smaller ones, as shown in Table 1. How cervigrams comply with general rules of graphic presentation is shown in Table 2. Labeling of axes is done on 4 of 22 axes and unambiguous labeling of divisions on scales on 9 of 22 scales. Slope of the line of dilatation for dilatation rate of 1 cm/h in one cervigram is approximately 45 degrees (optimal slope). In the remaining 10 cervigrams the slope is significantly smaller being in the range from 25 to 30 degrees. Possibility of precise entering of data can be seen in Table 3. Dimensions of 1 cm x 1 h fields in all cervigrams except one are not integers. Smallest units on time scales are between 0.25 and 2 h. DISCUSSION: Unlabeled coordinate axes, which are almost the rule for cervigrams in Serbia can be seen on other cervigrams [7]. However, it may not be of importance since cervigrams are mostly used by those who draw them. Accuracy and precision of entering data into cervigrams depends on: unambiguous labeling of scale divisions, dimensions of 1 cm x 1 h fields and smallest units on scales. Smallest units on time scales of cervigrams presented by other authors [8-10] are at most 1 h, while cervigrams in Serbia in 7 of 11 have smallest units of 2 h. Unambiguous labeling of scale divisions on cervigrams of the same authors is not noticed. Cervigrams in Serbian maternity hospitals exhibit unambiguous labeling in 13 of 22 observed scales. Since space for cervigram is often determined by the number of other parameters and space they occupy [8], dimensions of 1 cm x 1 h fields in mm are frequently not integers. This is the reason why values smaller than the smallest units on scales can be entered with great difficulty, if at all. Ten of eleven cervigrams from maternity hospitals in Serbia show such characteristics. CONCLUSION: Cervigrams which are being used in maternity hospitals in Serbia show significantly lesser slope for 1 cm/h dilatation than 45 degrees and dimensions of 1 cm x 1 h fields in mm which are not being integers in all except in one cervigram for each characteristic. Unambiguous labeling of divisions is present in less than half (9 of 22) of scales and smallest units on time scales in most (7 of 11) of them are 2 h. Construction of cervigrams in such a manner does not provide accurate and precise graphic presentation of dilatation during labor and can be the source of error in obstetric decision-making. Lesser than optimal slope for 1 cm/h dilatation may influence obstetricians decisions towards more interventions during labor when such interventions are not necessary. PMID- 11928602 TI - [Diagnostic value of hysterosalpingography in examination of fallopian tubes in infertile women]. AB - Hysterosalpingography (HSG) is a radiographic examination of endocervical canals, uterine cavity and Fallopian tube with the use of a radiographic contrast medium [1]. This method is an integral part of gynaecological examination and its value has not been underestimated in the modern gynaecological practice. GOAL OF THE STUDY: The goal of the study was to evaluate the reliability of HSG in the diagnosis of Fallopian tube and to compare the obtained results with laparoscopic findings. METHODS: The study included 140 infertile women. HSG was performed in the first half of the cycle, usually on the ninth day, without anaesthesia. The instruments after Schultze were used; 15 mL of Telebrix-contrast was used. Three radiograms were done. Laparoscopic examination was carried out in general endotracheal anaesthesia. A Storz laparoscope was used. CO2 was used for artificial pneumoperitoneum and indigolipstick for tube passage. The obtained findings were elaborated statistically. Descriptive and analytic models were used. p < 0.05 and p < 0.01 were considered as a risk factor of statistical significance. RESULTS: An approximate time interval between the two procedures was 5.18 months. Normal findings of HSG examination were noted in 53 women (37.9%); tube occlusion in 67 women (47.9%), and peritubal adhesion with tubal passage in 20 (14.3%) patients. A normal finding was found in 56 women (40.0%), tubal occlusion in 64 women (45.7%), and peritubal adhesion with tubal passage in 20 (14.3%) patients. HSG and laparoscopic findings regarding normal tubes were in agreement in 32 women (22.9%), tubal occlusion in 35 women (25.0%) and peritubal adhesion with tubal passage in 5 (3.6%) patients. The best sensitivity of HSG was observed in detection of proximal tubal occlusion (78%), and the smallest in occlusion with the accompanying adhesion (2%). The best specificity of HSG was noted in the diagnosis of combined occlusions (96%), and the smallest in tubal passage with peritubal adhesion (25%). There were 15% of false negative findings and 17.1% of false positive findings. DISCUSSION: The time interval from one to the other procedure can be considered as an important factor in laparoscopic confirmation or negative HSG findings. With the continuation of the same time interval the conditions are made for the aggravation of old and occurrence of new pathological processes in genital internal female organs. The possible causes of differential diagnosis of tubal occlusion between HSG and laparoscopic examination might be: 1) unequal anaesthesia during HSG and laparoscopic examination; 2) different properties of contrast media; 3) anatomic variations in the width of lumen tubes; 4) erroneous interpretation of the results. The sensitivity of HSG in this study was different in various types of tubal passage. In other studies the sensitivity of HSG was from 65% [10] to 96% [7]. The high specificity was found during detection of combined tubal occlusion (96%). The results of other authors were similar [7, 10]. This is a good contribution to the statement that HSG is a useful test of tubal obstruction. A rather high percent of false positive results of HSG was established in this study. The possible reasons might be tubal spasm and endometrial polyp in the area of the uterine opening of the tubes. CONCLUSION: On the basis of the obtained results, the following conclusions can be drawn: 1) HSG is a simple method for examination of female sterility; 2) HSG and laparoscopy are the complementary methods in the examination of tubal sterility; 3) HSG is inferior in relation to laparoscopy in the examination of peritubal adhesion. PMID- 11928603 TI - [Soft tissue injuries of the neck]. AB - Thirty five patients were examined. All had soft tissue spinal cord injury and were not treated in a hospital. Complete diagnostic and therapeutical protocol was carried out only for 13 patients. The goal was to compare their initial condition with condition after the control examination after the therapy. Results showed that 8 patients (23%) suffered from pain in the neck, 25 (72%) had limited neck mobility, and 17 (49%) manifested neurological symptoms. Signs of osteochondrosis at isolated spinal levels and dyscoligamental instability were noted in one fifth of the observed patients. In 24 patients (69%) the condition was defined as "serious". Although soft tissue spinal cord injuries are considered as "light" injuries, they can cause serious consequences that reduce the quality of patient's life due to inadequate treatment. From that point of view, a serious approach to diagnosis and treatment is necessary. PMID- 11928604 TI - [Multifocal localization of tumors of Schwann cell origin]. AB - Neurofibromatosis (disease of Von Recklinghausen) was considered to be a unique disease. Today, however, it is known that it consist of two clinically and genetically different diseases. Neurofibromatosis type 1 is a more frequent disease formerly known as peripheral, while neurofibromatosis type 2 (bilateral acoustic neurinoma) is significantly less frequent. In this case report we present a 30-year old female patient who had several tumours with the following localizations: 1. the pontocerebellar angulus; 2. the left foramen jugular; 3. the anterior part of the spinal canal; 4. the soft tissue of the neck; 5. falx cerebri. The tumor in the neck was removed completely by ENT surgeon while the others were partly removed by neurosurgeon. In this case report we described diagnostic and therapeutic problems in the treatment of this patient with the long-lasting hoarseness and difficult swallowing. PMID- 11928605 TI - [The Mallory-Weiss syndrome as an unrecognized cause of death]. AB - Longitudinal tears in the esophagus at the esophageogastric junction are termed Mallory-Weiss syndrome. They are encountered most commonly in alcoholics, attributed to episodes of excessive vomiting. These lacerations could be the cause of massive and severe external and/or internal fatal bleeding. Bleeding from upper gastrointestinal parts could be caused by this syndrome, as well as by ulcers, inflammations, esophageal varices, tumours, blunt abdominal injuries, etc. Such unclear deaths could be of interest to law authorities because they are suspicious, sudden and/or obscure. In daily forensic medical practice, this syndrome is relatively rare, and therefore it, it should be recognized. In this paper, the authors present the case of a single male, aged 54, a chronic drinker, who was found dead in his flat. During the previous day, he was observed medically in the Trauma Centre in Belgrade for nonspecific gastrointestinal symptoms (nausea, vomiting and diarrhea). The mucous tears of esophageogastric junction had been established by autopsy, as well as the massive internal gastrointestinal bleeding. On the basis of autopsy and microscopic findings, clinical data and circumstances, the established cause of natural death was fatal exsanguination from esophageogastric mucous tears due to Mallory-Weiss syndrome. PMID- 11928606 TI - [The Lesch-Nyhan syndrome]. AB - Deficiency of hypoxanthine phosphoribosyltransferase (HPRT) has a broad spectrum of clinical manifestations, from the complete enzyme defect, the Lesch-Nyhan syndrome with severe neurological deficiency to the partial defect associated only with uric acid overproduction and its consequences. We present a 5-year old boy with Lesch-Nyhan syndrome. He came to our hospital because of abdominal pain, vomiting and gross haematuria. At the age of 8 months he was categorized as a "cerebral palsy" patient due to involuntary movements and high degree of spastically and tonic spasms. He remained incapable of sitting or standing alone. The patient's brother and two uncles were also categorized as "cerebral palsy" cases and died at the age of 8-14 years. Clinical examination revealed hyperuricaemia and hyperuricosuria, radiolucent renal and urinary bladder stones. HPRT enzyme activity was totally absent, while adenine phosphoribosyl transferase activity was increased compared to control. The patient was treated with allopurinol, urinary alkalization, low-purine diet and adequate hydration while he was in hospital. However, his parents refused further treatment and follow-up. The most important issue is whether the healthy sisters of the patients are heterozygotes for HPRT deficiency. This DNA analysis is now in progress. PMID- 11928607 TI - [Retropancreatic hematoma mimicking a tumor in a female patient with liver amyloidosis]. AB - Bleeding in patients with amyloidosis is considered as a frequent complication appearing in up to 40% of cases. It may be spontaneous or provoked. Spontaneous bleeding in the skin or in the gastrointestinal tract is the most frequent. Spontaneous bleeding in the retroperitoneum is rare. It may be diffuse, with possible lethal outcome, or localised, causing haematoma, as in our patient. We present a 62-year old woman with a three-month history of spontaneous, subcutaneous bleeding. No laboratory or other findings could explain the cause of bleeding, as the haemostatic tests were within or close to normal limits. US and CT performed for right subcostal pain discovered a tumour attached to the back of the head of pancreas. Due to mild coaugated hyperbilirubinaemia the "tumour" was held to be the cause of patient's difficulties. However, at operation it turned out to be a retropancreatic haematoma, which was removed and drained. Because of conjugated hyperbilirubinaemia cholecystectomy with operative cholangiography (which was normal) were performed. Due to steatotic appearance of the liver a biopsy was performed. Histological examination revealed amyloidosis. The recovery was uneventful and the patient was transferred to hepatology unit for further treatment where she died 3 months later due to gradual deterioration of kidney and heart function. To our knowledge, retroperitoneal haematoma in liver amyloidosis mimicking retropancreatic tumour has not been reported so far. PMID- 11928608 TI - [Radiotherapy in combined therapy of medulloblastoma]. PMID- 11928609 TI - [The evidence-based system in oncology]. PMID- 11928610 TI - [135 years' since the first medical textbook for higher education in Serbia]. AB - Medical teaching at School in Belgrade started in 1863 when forensic medicine became an oblicatory subject for law students. In 1865 the Serbian physician Joachim Medovitsh (1815-1893) published the first medical textbook "Forensic medicine for lawyers" in Serbia. It appeared more than fifty years before foundation of the School of Medicine in Serbia. The book appeared forty years before promoting the High School to University. Joachim Medovitsh was of Polish origin but he lived and worked in Serbia. He was one of the most prominent physicians in Serbia and the first president of the Serbian Medical Society. Joachim Medovitsh, M. D. wrote his book after the model of the textbook of Karl Bergman (1814-1865). German anatomist and physiologist from Gottingen. Forensic medicine of Medovitsh was printed by lithographic technique. The second edition was edited next year. In his book Medovitsh cited often Johann Casper (1796-1864) one of who published the most used textbook in the second half of the 19th century. Joachim Medovitsh, M. D. contributed very much to the beginnings of High School medical teaching in Belgrade being in the course of events and achievements of European medicine. PMID- 11928611 TI - [Triumph of democracy in the Serbian Medical Society]. PMID- 11928612 TI - [The yentl syndrome in women with arterial hypertension and positive findings on the echocardiography dipyridamole stress test]. AB - The aim of this study was to assess the possible gender differences between positive (new wall motion abnormality) combined dipyridamole stress echocardiography and coronary angiography or myocardial revascularization procedures. We evaluated 127 consecutive patients (82 males, 53.5 years old and 45 females, 56.1 years old) with systemic hypertension and chest pain (angina or atypical chest pain) who underwent pharmacological stress echocardiography (dipyridamole-atropine or dipyridamole-dobutamine) for detection of myocardial ischaemia. Coronary angiography was carried out in 84 patients (58 males and 26 females). Diagnostic accuracy of the test was the same in women as in men (92.3 vs 89.1%, females vs males, respectively). Coronary angiography was done in 73.6% of male and 66.7% of female patients with positive test; the difference was not significant. Over a two-year follow-up there were 31 myocardial revascularization procedures (20 coronary surgery and 11 angioplasty), 25 in men and 6 in women (p = 0.028). There was a trend in women with positive test to have less surgery or angioplasty (28.6%) than in men (45.3%), but the difference was not significant. Cox multivariate regression analysis revealed that women were less likely to receive revascularization (relative risk for females RR = 0.1457, 95% CI 0.0416 0.5101). CONCLUSION: Although diagnostic accuracy of third generation dipyridamole stress echocardiography was not different between men and women, in patients with positive test there was a trend in women to undergo coronary angiography and coronary surgery or angioplasty in lesser degree than in men. Cox multivariate model showed low relative risk of females for revascularization procedures and possible Yentl's syndrome in studied population. PMID- 11928613 TI - [Correlation between survival time and severity of injuries in fatal injuries in traffic accidents]. AB - INTRODUCTION: In forensic pathology, only trauma systems based on disintegration of anatomic structure of organs and tissues, could be used for objectivization, comparison and establishing of severity of injuries. Trauma systems based on pathophysiological values are useless. The Abbreviated Injury Scale (AIS) and its derivate Injury Severity Score (ISS) are the most common. AIS coded injuries are divided into six body regions and injuries are assigned a six-digit score in relation to their severity. ISS results the sum of the squares of the highest AIS values from the three most severely injured body regions. In this way, the ISS values are discontinued and vary from 0 (absence of injuries) to 75 (incompatible with-life injury). PURPOSE: The purpose of this paper is to establish the correlation degree between outliving period and trauma severity in persons fatally injured in traffic accidents, and according to this finding to point out the ISS value of critical injury. MATERIAL AND METHOD: A retrospective autopsy study was performed; it included the material of the Institute of Forensic Medicine in Belgrade of 1998. The autopsy reports and accessible clinical medical data were analyzed for persons over the age of 18, fatally injured in traffic accidents who survived trauma less than 15 days. The sample was statistically prepared (chi 2-test, t-test, correlation coefficient, regression line). RESULTS AND DISCUSSION: The sample included 272 persons: 193 males and 79 females. The proportion of men was more significant (chi 2 = 4.76; 0.01 < p < 0.05). Average age was 51.08 years (SD = 18.08): males 49.84 +/- 17.41 and females 54.09 +/- 19.38. The most frequently injured persons in our sample were pedestrians (134). The authors combined the autopsy and accessible clinical data in order to obtain the ISS value for each case. They considered that all persons found dead on the spot or died ante portam did not outlive trauma. The sample distribution by ISS values showed three peaks: for ISS--75, 41-50 and 26-35. Peaks indicated the number of the injured body regions and trauma severity in these persons. In 87 persons who did not survive, the ISS value was 75. There were 73 persons without outliving period with ISS values less than 75: their mean ISS value was 31.87 (SD = 11.30). In 112 cases the mean outliving period was 4.79 days (SD = 3.77) and their mean ISS value was 18.05 (SD = 15.33), which was a statistically significant lower ISS value than in previous group (t = 7.015; p < 0.001). A weak negative correlation between outliving period and ISS values in our sample was noted (coefficient of linear correlation r = -0.452). Our sample is representative (t = 8.37). Coefficient of a determination (r2 = 0.20), pointed to the fact that direct correlation outliving period-trauma severity was only about 20% and the rest of correlation i.e. 80% depended on other factors (e.g. effective emergency medical system and triage, prompt and correct diagnosis, adequate medical treatment and care, etc.). The calculated linear regression was as follows: outliving period approximately 52-3 ISS. This regression pointed out that critical and potentially fatal injury, in our sample, was injury with ISS of 17. There were 22 persons with ISS < or = 7. Six of them died on the spot as car passengers; they died due to mechanical asphyxia (thoracoabdominal pressure) or respiratory and/or circulation failure due to critical chest injury (flail chest, contusions and rupture of the lungs with consequent haemopneumothorax). The rest of 16 persons survived trauma in an average of 8.56 days (SD = 3.88), and the causes of death were pneumonia, thrombus and fat embolism, sepsis, etc. CONCLUSION: By anaylzing our sample of fatally injured persons in traffic accidents (unpenetrated blunt trauma), there was a negative weak correlation between the outliving period and severity of injury based on ISS. This correlation was partly direct but mostly depended on other factors (e.g. effective emergency medical system and triage, prompt and correct diagnosis, adequate medical treatment and care, etc.). Establishment of these factors could be possible through state medical projects in big medical trauma centres. Prospect registration, evaluation and scoring of all injuries in hospitals and dissecting rooms, and comparison of the obtained results, can give valid data on mortality of injured people, bad diagnosis, and appropriate medical treatment. The autopsy of injured persons dead on the spot can point out what kind of injuries are incompatible with life, as well as with their severity. The autopsy of injured persons who survived trauma can point to the most frequent injury complications, clinical diagnosis and preventable deaths. According to this paper, the critical injury by ISS is 17. In such cases, the forensic pathologist must answer the following questions: whether the death was due to trauma; whether the precipitated cause of death was the consequence or complication of injury; what were the mechanism and mode of dying; whether the death was preventable; if there were possible malpractice and negligence, etc. PMID- 11928615 TI - [A double abscess of the cerebellum of otogenic origin]. AB - Brain abscess is a rare, extremely aggressive lethal infection. It is especially important in case of otogenic abscesses. It is known that the problem of otitis existed already thousands of years before Christ, and it is still a current problem. Otogenic brain abscess is the most common otogenic complication encompassing 40-80% of all brain abscesses in adults. In 50-80% of these cases COM (chronic otitis media) with cholesteatoma is found. In the last 20 years CT is of the first rank in diagnosis. Brain abscess can be treated conservatively and surgically; but the ear has to be treated operatively. In this article we present an interesting case from our practice, a double abscess of the cerebellum of otogenic origin, which was successfully treated by simultaneous approach of neurosurgeon and otosurgeon. PMID- 11928614 TI - [Encapsulated filtering blebs--incidence and methods of treatment]. AB - Encapsulation of the filtering bleb occurs as a complication of glaucoma filtering surgery in 8.3-28% of all eyes filtered, often between 2-4 weeks after surgery. It has a characteristic clinical appearance--highly elevated localized bleb, prominent surface vessels and patent sclerostomies on gonioscopy. This study retrospectively reviewed the results of 100 filtering operations, performed in the Prof. Dr. I. Stankovitsh Eye Department, focused on the incidence, possible risk factors and management of encapsulated filtering blebs. Trabeculectomy was done in all patients with limbus-based conjunctival flap, rectangular scleral flap 4 x 4 mm, and sclerostomy 2 x 2 mm. Two or three interrupted 10-0 nylon sutures were used to tether the flap, the conjunctiva with Tenon's capsule was closed with a running suture. All eyes received topical dexamethasone drops, mydriatics-homatropin or tropicamide and antibiotic ointment for three weeks. Possible risk factors were considered: sex, age, other eye or systemic disease, type of glaucoma, previous ocular procedure and antiglaucoma medical therapy, early postoperative complications, postoperative and final intraocular pressure (IOP). All eycs with encapsulated filtering bleb were given topical beta-blocker initially and carbonic anhydrase inhibitor, and if IOP continued to be uncontrolled incisional surgical management was performed. Success was defined as IOP maintained at less than 22 mmHg, with or without medication. An encapsulated filtering bleb developed in 9% of eyes. Previous argon laser trabeculoplasty was associated with an increased frequency of bleb encapsulation. The mean postoperative IOP was 11.30 +/- 1.90. Encapsulation of filtering bleb was developing at mean time of 19.9 +/- 3.0 after surgery. Six eyes returned to IOP below 21 (mean IOP was 20.17 +/- 0.90) within 4 weeks, and 4 of them continued on a topical beta-blocker. Four eyes required incisional surgery. A week after surgery mean IOP was 18 mmHg. Haemophthalmus occurred after one incision performed in a patient suffering from diabetes mellitus. Late scarring of the filtering bleb was developed in 2 eyes. Seven months and 2 years after filtration surgery mean IOP was 21.30 +/- 4.00 mmHg. The cause of bleb encapsulation is not known. Male patients, surgical glove powder, topical corticosteroids, previous argon laser trabeculoplasty and beta-blockers are some of potential risk factors for the development of encapsulated filtering bleb. This study suggests the association between development of encapsulated bleb and previous argon laser trabeculoplasty--40% in the eyes with encapsulation and 18% in the eyes without encapsulation. The optimal management of bleb encapsulation has not been defined. In this study topical antiglaucoma therapy achieved IOP control and cyst's remodeling in 60% of cases. Forty percent of all cysts required incisional surgical management. Encapsulation of the filtering bleb is not an uncommon complication of glaucoma filtration surgery. It is important to examine postoperative eyes frequently during the first 2 months to detect this complication. Fortunately, most eyes have their IOPs controlled with antiglaucoma therapy. PMID- 11928616 TI - [Successful treatment of primary myelofibrosis with thrombocytosis during pregnancy with alfa-interferon]. AB - Primary myelofibrosis is predominantly a disease of old age, poor prognosis and no curable treatment. Thrombocytosis was observed in only 12% of patients. To our knowledge, there is only one reported case of a young woman with primary myelofibrosis who had a term pregnancy [1]. We report on a 29-year-old woman with thrombocytosis and medical history of two miscarriages in the last 2 years, the iirst at 30 weeks of gestation and the second at 27 weeks. Multiple placental infarctions were observed. She was without symptoms but with moderate splenomegaly 4.5 cm below left costal margin). The platelet count was 651 x 10(9)/L, WBC 7.2 x 10(9)/L with normal differential formula, and haemoglobin level 12 g/dl. Bone marrow biopsy showed fibrotic phase of primary myelofibrosis, with hyperplasia of megacaryocytes, decreased numbers of erythroid and granulocytic cells, and increased amounts of reticulin fibres. Cyctogenetic examination of the bone marrow showed normal female caryotype. Increased numbers of progenitors CFU-Mk, CFU-GM and BFU-E were observed in peripheral blood, and decreased numbers in bone marrow cultures. As the patient wished to become pregnant, the treatment with interferon-a (Roferon A) was started at a dose of 3 MU s.c., three times per week. The platelet count rapidly decreased at a level of 260-370 x 10(9)/L. The pregnancy was diagnosed 5 months later. At the 24 week of pregnancy, platelet count raised to 690 x 10(9)/l and the dose of interferon-a was augmented, 3 MU every day, until delivery. Foetal growth and placental circulation were monitored by serial ultrasonography. At the end of 34 weeks of pregnancy, it was noted that placental flow became insufficient, and after foetal lung maturity was stimulated with dexamethasone, Cesarean section was performed. Male baby was born, weighting 2000 g, with respiratory distress syndrome. This complication was successfully treated, and the child is now one year old, with normal growth and development. The mother is still on therapy with interferon-a, 3 MU, three times a week, and the last blood count was as follows: haemoglobin 10.7 g/dl, WBC 6.1 x 10(9)/L and platelet comt 437 x 10(9)/L. In conclusion, according to the clinical results of interferon-d in thrombocytosis and experimental studies which showed the absence of placental transfer of interferon d, this therapy could be recommended to women with primary myelofibrosis who wish to have a baby. PMID- 11928618 TI - [Functional morphology of the hippocampal region in higher primates]. PMID- 11928617 TI - [A retroperitoneal bronchogenic cyst]. AB - Bronchogenic cyst is a rare congenital (developmental) anomaly. It is usually asymptomatic but its enlargement and localization may cause serious symptoms. Exact preoperative diagnosis is rarely established. As a rule, it is established during histological examination of the resected specimen. We present a patient with a subdiaphragmatic retroperitoneal bronchogenic cyst in whom exact diagnosis was documented by histological examination of excised cyst. To our knowledge this is the third such case ever reported. PMID- 11928619 TI - [Etiology and pathogenesis of alopecia areata]. PMID- 11928620 TI - [Pancreatic carcinoma: from genes to therapy]. PMID- 11928621 TI - [Current knowledge of mycoplasmas]. PMID- 11928622 TI - [Primary B-cell cutaneous lymphoma: a model for study of lymphogenesis associated with a specific infectious agents]. PMID- 11928623 TI - [Authorship in medical articles]. PMID- 11928624 TI - Common mechanisms in the damaging effects of hypoxia and hyperglycemia on neuronal function. PMID- 11928626 TI - [Humoral autoimmune responses to neurospecific proteins in rats with experimental allergic encephalomyelitis and immunomodulation with allogeneic nervous tissue]. AB - Possible immunocorrective effect of the embryonic nervous tissue (NT) and using the model of an experimental allergic encephalomyelitis (EAE) in treating patients with demyelinizing diseases is a prospective field of research. EAE was induced by immunization of rats with the mixture of the spinal cord and BCV at paw pillows. Immunocorrective effect of the allogenic NT was studied in the animals with EAE following the intraperitoneal injection of a newborn rat's NT, namely: enriched fractions of glial and neuronal cell populations. Both intact and EAE animals, treated with culture medium, were used as controls. The clinical signs appeared on 12-th day after EAE inducing. The severe clinical course of EAE was accompanied with statistically significant higher titers of the autoantibodies to MBP as compared with the mild clinical course. The results obtained evidence for the possible immunocorrective effect of enriched fractions of the newborn rat's nervous cells on EAE, and for immunosuppressive effect of the neuronal fraction of NT on the immunopathological processes in EAE. PMID- 11928625 TI - [Current concepts on the mechanisms of hypoxic effects on vascular tonus]. AB - It is well known that hypoxia causes smooth muscle relaxation of the majority of mammalian systemic blood vessels, whereas smooth muscles in the pulmonary and large coronary arteries constrict under hypoxia. The review describes a modern concept of the mechanisms involved in the hypoxic vasoconstriction and vasodilatation. Cationic channels of a plasma membrane, the contractile apparatus, and mitochondria are the main oxygen sensors in the vascular smooth muscle cells. Hypoxic vasodilatation is mediated mainly by a decrease in the voltage-dependent Ca2+ entry, decrease in Ca2+ sensitivity of the contractile apparatus, and activation of ATP-dependent K+ channels. This process also involves endothelium derived nitric oxide. Hypoxic vasoconstriction mechanisms may be related to voltage-gating K+ channels inhibition, Ca2+ release from intracellular stores and inactivation of Ca2+ activated K+ channels each of them leads to increase in intracellular Ca2+ concentration. Platelet-activating factor, prostaglandins F2 alpha, E2, tromboxan B2, leucotriens C4 and D4 also contribute to hypoxic vasoconstriction. Glycolysis which intensity increases in hypoxia, and electron transport chain which generates the reactive oxygen species play the important role in the development of hypoxic pulmonary vasoconstriction. They possess the ability to change redox state in the cells and therefore to modulate the activity of the cationic channels. Hypoxia also leads to a proliferation of smooth muscles in the vascular wall. Better understanding of the underlying hypoxia-related mechanisms is vital for the explanation of enhanced blood flow under hypoxia, and is absolutely necessary for creating new effective antihypoxic drugs. PMID- 11928627 TI - [Effects of adrenoreceptor agonists and cyclic nucleotides on membrane potential of isolated endothelial cells of the rabbit aorta]. AB - The influence of adrenoceptor agonists and changes in the intracellular cyclic nucleotide concentration on the membrane potential of an isolated endothelial cell layer from rabbit aorta has been investigated. The alpha 1-adrenoceptor agonist phenylephrine evoked deep depolarization of the endothelium of an intact aorta, but had a little effect on the membrane potential of the isolated endothelium. The nonselective adrenoceptor agonist noradrenaline, the beta adrenoceptor agonist isoproterenol and the inhibitor of phosphodiesterases IBMX all strongly depolarized the isolated endothelium. These data suggest that the rise in cyclic nucleotide concentration depolarizes endothelial cells, and this fact may account for the noradrenaline-induced depolarization of the endothelium through beta-adrenoceptors. PMID- 11928628 TI - [Study of a factor released during myocardial ischemia reperfusion and its effects on myocardial, coronary and peripheral vessels]. AB - The effect of reoxygenated coronary effluent of an isolated guinea-pig heart on a sequentially perfused second heart, right auricle trabecula and arterial vessel rings was studied after 20 min ischaemia of the first heart. It has been shown that a factor released after myocardial ischemia/reperfusion stimulates arrhythmia, decreases myocardial contractility of the second heart and depresses tonical tension of the right auricle trabecula and arterial vessel rings. Storage of the coronary effluent up to 24 h did not modify its effects. These results suggest stable factor released from an isolated heart after ischaemia at reperfusion to exert humoral effect not only on a myocardial contractility and coronary circulation but on the peripheral vessels tonus. PMID- 11928629 TI - [Pathogenetic role of prostanoids in disorders of sodium reabsorption during development of acute renal insufficiency]. AB - The experiments were carried out on 36 male rats with an acute renal insufficiency--sublimate-induced nephropathy. An increased amount of both tromboxan A2, and prostaglandine F2a has been shown to play a pathogenic role in the disorders of the main energy-dependent process in kidney--reabsorbtion of sodium ions. The negative correlative relation between the concentration of sodium ions in plasma and an increased amount of prostaglandine E2 in a kidney papilla can be explained by the hyperfunction of the interstitial cells of a kidney papilla and with a natriuretic effect of prostaglandine E2 at the level of collective tubules. PMID- 11928630 TI - [Disturbances in biliary secretion during tissue hypoxia and attempts at their corrections]. AB - In acute experiments on rats in vivo the stimulating effects of hydrophilic cholic acids (CA and UDCA) on the bilification and their inhibiting by rotenone have been shown. The secretion of CA and UDCA under rotenone have been determined to be activated. The mechanisms of the influence of the cholic acids on the secretory function of the hepatocytes have being discussed. It has been concluded that NADH-dehydrogenase link in the respiratory chain is not involved in the activating effects by CA and UDCA. PMID- 11928631 TI - [Thrombocyte function during normal and complicated pregnancies]. AB - Research of functional changes in the circulating thrombocytes at physiological pregnancy and EPH--gestosis in women was conducted with methods of an electron microscopy and microcoagulogy. Some peculiarities of the release reactions by thrombocytes which determine an extra thrombocyte release of the content from their cytoplasmatic granules have been determined. PMID- 11928632 TI - [Changes in energy metabolism in the brain of bemegride-kindled rabbits]. AB - Changes in the energetic metabolism in the cortex were examined in bemegride kindled rabbits. A reliable decrease in the ATP and an increase in the AMP contents in the cortical tissues as well as low ATP/AMP and energetic charge coefficients have been found after kindling the rabbits for 14 and 30-days as compared to the control groups of animals. Considerable disorders in the oxidative phosphorilation in the mitochondria of the cortical tissues have been determined after the succinate and glutamate oxidation: a decrease in the oxygen utilization rate at V3 active metabolic state, lowering the respiratory control coefficient and the disscciation between phosphorilation and oxidation. Those disorders were more expressed after glutamate oxidation as compared to a succinate one. In addition, it has been determined that oxygen utilization at V2 state was reliably decreased; both coefficient of energetic production and ATP ase reserve activity lowered. PMID- 11928633 TI - [Role of interleukin-1 and interleukin-2 in the pathogenesis of systemic scleroderma]. AB - The purpose of a research was to define the levels of interleukin-1 (IL-1) and interleukin-2 (IL-2) in patients with systemic scleroderma (SSc) and to analyse their dependence on the course of the illness. 75 patients with SSc and 20 apparently healthy people were examined. Besides the well-known clinical biochemical and instrumental investigations, the levels of IL-1 and Il-2 in all the patients were determined in serum with an immunoenzymatic method using a set of reactants Pro Con IL-1b and Pro Con IL-2 (Russia). The data obtained give evidence for an increase in both IL-1 and IL-2 levels. As the correlation between the level of IL-1 and the acuteness of the pathological process has been shown to be especially distinct, it can be used for marking an acute form of this pathology. Thus, both parametres (IL-1, IL-2) depend on the acuity and peculiarities of SSc, and on the concomitant damage of the internal organs. The most pronounced increase in LL-2 level has been observed at visceralisation of SSc. PMID- 11928634 TI - [Effect of restored 5alpha-androgens on puberty in female rats]. AB - The female rats at a prepubertal period were introduced to the restored 5 alpha androgens, namely: 5 alpha-dihydrotestosterone (DHT), 5 alpha-androstane-3 alpha, 17 beta-diol (3 alpha-diol) or 5 alpha-androstane-3 beta, 17 beta-diol (3 beta diol) in the dose of 5 mg/100 g. Both DHT and 3 alpha-diol have been determined to induce the vaginal opening as the initial stage of puberty. Stimulation of the puberty in those rats was followed with an activation of both endocrine and generative functions of the ovaries. DHT inhibited the gonadotrophic activity of the hypophysis that delayed the ovulation as the final stage of puberty. The DHT metabolite 3 alpha-diol hastened the first ovulation, while 3 beta-diol did not influence the pace of the puberty. The stimulating effect of DHT at the initial stage of the pubescence in the female rats has been suggested to be due to its conversion into 3 alpha-diol which is likely to be one of the key factors in the pubescence control in rats. PMID- 11928635 TI - [Dynamics and dose-dependent changes in RNA and DNA synthesis in the thymus of irradiated animals]. AB - Effects of an exposure of male vistar rats to 60Co--quanta in the doses of 0.2; 0.5; 1.0; 3.0; 5.0 Gy for 120 days; 7.0 Gy for 20 days and 9.0 Gy for 15 days on the dynamics of the thymic nucleinic acids have been studied. Experiments were conducted in 0.5, 1, 2, 4, 6, 8, 10, 15, 20, 30, 45, 65, 90, 120 days after an exposure to radiation. Essential shifts in metabolism of the nucleinic acids determined on the base of changes in their contents and ratio RNA/DNA have been established. The mass reduction of a radiosensitive organ was mainly due to a decrease in its cellular tissue. The greatest rate of RNA and DNA changes has been found to be on the first day following radiation. Significant reduce in the thymic total DNA synthesis can testify to a sharp decrease in the cellular production by this organ, and the decrease in the RNA synthesis gives evidence for a delay of the differentiation of the thymic cells. Defining the ratio of DNA to RNA can serve a qualitative indicator of the additional generations of the proliferating cells and finally of the size and state of trunk cells. PMID- 11928637 TI - [Age-related characteristics of experimental hypothyroidism in rats]. AB - The rate of both the synthesis of liver and plasma proteins and RNA-1 and RNA-2 polymerase activities in liver were studied in rats of various ages at experimental hypothyroidism. There has been marked more significant decrease in plasma protein synthesis with age. Both the rate of liver and plasma protein synthesis have been shown to be reduced at experimental hypothyroidism but synthesis of plasma proteins was inhibited to a greater extent. Considerable changes were observed neither in liver and plasma protein synthesis nor in the balance between these two groups of the protein synthesis in old rats. RNA-1 and RNA-2 polymerase activities decreased at hypothyroidism. At all ages the activity of bound enzymes decreased to a larger extent as compared to free forms. The activity of RNA-polymerase 2 was more inhibited than that of RNA-polymerase 1. Reducing protein- and RNA-synthetic processes in the liver with age correlated with the peculiarities of the carbohydrate metabolism: in those young animals with an impaired glucose tolerance the inhibitory effect of hypothyroidism on the intensity of protein and RNA synthesis was more potent as compared to old animals. PMID- 11928636 TI - [The effect of hypergravity on antioxidant enzymes in rat tissues and blood]. AB - The influence of hypergravity (5, 8, 12 N.m2/kg2 for 15 min) on the activity of the antioxidant enzyme catalase and superoxide dismutase has been investigated in the blood, brain and liver tissues of rats in 15, 30, 60 min., and in 1, 3, 7 days after loading. It has been shown that peculiarities of the physiological response to the hypergravity of different intensity depend on the intensity of loading and functions of the tissues under investigation. PMID- 11928639 TI - [Functional state of the central nervous system in people with varying functional agility of the nervous processes while transforming information of varying complexity]. AB - Relations between functional agility of the nervous processes (FANP) and functional state (FS) of the central nervous system at performing simple and difficult sensomotor tasks have been studied. It has been shown that FANP is of the exceptional importance in forming FS of the central nervous system only to perform difficult mental tasks. PMID- 11928638 TI - [Intensity of lipid peroxidation and antioxidant enzyme activity in arterial and venous walls during hypervitaminosis D]. AB - The intensity of the lipid peroxydation (LPO) and the antioxidant enzyme activity (superoxide dismutase, glutathione peroxydase and catalase) on injecting vitamin D in high doses (10,000 U/kg) was examined in the arterial and venous walls of rabbits. The increase in the amount of the intermediate and final LPO products has been found in the vessels of all types. The lowest intensity of LPO was noted in the vena cava. The decrease in the antioxidant activity has been revealed. But vena cava inferior was the exception because the activity of all studied antioxidant enzymes grew in its wall. This increase is likely to be one of the reasons for vena resistance to the action of damaging factors. PMID- 11928640 TI - [Age-related effects in thymocytes differentiation: role of pineal gland peptide factors]. AB - The peculiarities of the differentiation of the thymocytes in adult and old CBA/Ca mice were studied with a help of the monoclonal antibodies to different lymphoid antigens of mice. In the thymus of old mice, the portions of the CD4-CD8 , CD44+ and CD44+Thy-1(-)-cells have been shown to be increased. The percentage of the CD4+CD8(+)-thymocytes did not change, whereas the portion of CD25+, PNA+, CD3+, CD3+CD8+, CD4+CD8- and CD4-CD8(+)-cells diminished. The ratios of different subpopulations of thymocytes, characterizing different degree of their maturation are changing with age towards the prevalence of less mature cells. After prolonged (for 18 months) administration of epithalamine the portion of CD44+, CD44+Thy-1- and CD3- 8(+)-cells in the old thymus has been shown to be decreased, while that of CD25+, CD3+, CD3+CD8+, CD4+CD8/and CD4-CD8(+)-cells increased. According to many parametres thymus of those old animals corresponds to adult mice. Epithalamine raised the titer of one of the thymic hormones--thymic serum factor, both in the blood of the old mice and in the supernatant of a 3-hour culture of the thymic stroma. PMID- 11928642 TI - E-newsletters bring visitors back to your site. PMID- 11928641 TI - [Effects of T-type calcium channel blockers on spontaneous meiotic maturation of mouse ovarian oocytes in vitro]. AB - We studied the effects of blockers of the T-type Ca2+ channels on resumption of meiosis and spontaneous meiotic maturation of murine ovarial oocytes in vitro. We found that the efficiency of suppression of oocyte maturation by blockers of the Ca2+ channels increases in the following succession: amiloride hydrochloride, mebifradil, NiCl2. It is supposed that there is a possibility to use blockers of the T-type Ca2+ channels for creation of new combined intrauterine contraceptives. PMID- 11928643 TI - Growing your e-newsletter subscriber list. PMID- 11928644 TI - Baptist Healthcare upgrades its physician access network. PMID- 11928645 TI - Major hospital buys ISPs (Internet service providers) for technological and marketing advantages. PMID- 11928646 TI - Economy, 9/11 tragedy send reinsurance market reeling. PMID- 11928647 TI - Conduct physician outlier analysis to gain control of costs. AB - Find out about a tool you can use to conduct retrospective profiling of physician practice patterns, identify outlying physicians, and unearth cost reductions under capitation. PMID- 11928648 TI - Nail down data on cost, utilization to ensure success of DM risk contract. AB - Many disease management (DM) companies are negotiating capitated rates for their services, but industry experts offer two major cautions for both DM companies and health plans: Nail down your cost per disease state and the prevalence of the disease within your covered population before even thinking about signing a contract. PMID- 11928649 TI - Inpatient days trending up after decade of decline. AB - Data Insight: The latest evidence of the impact widespread adoption of consumer choice models has had on health care utilization comes from Milliman USA's latest Intercompany Rate Survey. For the first time in nearly a decade, the survey shows a significant uptick in commercial inpatient hospital days per thousand. PMID- 11928650 TI - Automate claims processing to enhance profits under capitation. AB - If you're trying to find ways to wring more profit from your capitation contracts, reducing the costs and inefficiencies inherent in claims processing can help. Here's how. PMID- 11928651 TI - Michael Balint: an introduction. AB - Michael Balint, who used to be known the world over, no longer gets either the fame or the influence that he deserves, and the three special issues of the journal devoted to him are meant to contribute to make him better known. I intend to draw a portrait of Balint--it will necessarily be only an outline, within the limits of the present issue--and to follow his path as a man and as a scholar. PMID- 11928652 TI - Michael Balint then and now: a contemporary appraisal. AB - This paper pays tribute to Michael Balint's psychoanalytic contributions, thus far neglected in the United States. To assess the current relevance of some of Balint's pioneering ideas, the paper highlights some of Balint's core concepts. These gain in importance and enduring relevance when viewed from the perspective of Heinz Kohut's psychoanalytic self psychology. The "basic fault" and the unsurpassed depth and subtlety of Balint's description of the archaic transferences are examples of what are of lasting value in his work. PMID- 11928653 TI - Michael Balint: an overview. AB - Balint's important work extended over a period of 40 years in two directions; firstly the development of individual sexuality, and secondly, the development of object relationships, together with their relationship to psychoanalytical technique. He introduced the concept of primary love and was one of the pioneers in the thinking of the interdependence of mother and infant. He extended Ferenczi's work on therapeutic regression, introducing the concepts, among others, of benign and malignant regression, the basic fault, and the states of ocnophilia and philobatism. His applied psychoanalytic work is world famous, particularly the Balint groups for general practitioners and similar groups for marital therapists, social workers, and psychosexual counselors. PMID- 11928654 TI - Zones of maturation and regression, and psychoanalytic technique. AB - This article summarizes Balint's thoughts on the fundamental problems of psychoanalytic technique and their links with libidinal development as conceived by him. PMID- 11928656 TI - The crisis of medical practice. PMID- 11928655 TI - Balint after Lacan ... and after. AB - This paper examines Balint and Lacan's views about regression and symbolism, language, and transference. The author points out their similarities, their differences, and then proposes a synthesis through his approach of history, genealogy, trauma, and crisis. Freud's notions of masochism, of fixation (innate or phylogenetic) may thus be renewed, leading to the analyst's capacity to cope with deep regression and borderline (Balint) psychosis and perversion (Lacan) or to psychosomatics, including epilepsy. PMID- 11928657 TI - [Airway function disturbances reversibility after mitral valve replacement]. AB - Dyspnea, cough, recurrent airway infection, hemoptysis are the most common pulmonary symptoms of mitral valve disease and heart failure. Pathophysiological mechanism of those disturbances is complex and airway status is one of the most important. The aim of the study was to assess airway function disturbances reversibility after mitral valve replacement. The study group consisted of 30 patients qualified to mitral valve surgery. Patients were assessed by clinical cardiac noninvasive investigation and airway function study. Post-operative study was performed minimum 6 months after mitral valve replacement (mean after 8 months) and again after minimum 3 years (mean after 40 months). In most of assessed--22 patients (74%) airway obstruction was noticed, in 8 patients without obstruction nonspecific histamine provocation test was performed. Increased airway reactivity was found in 4 patients only, in another 4 patients (13%) there was no airway function disturbances. After mitral valve replacement significant improvement in all cardiac parameters including NYHA functional class was observed. No airway function improvement occurred. Only small tendency to improve airway function was noticed as far as it concerns VC, FEV1, MEF50, MEF75 iTGV with exclusion of Raw. Analysis after dividing study group into 3 subgroups with increasing airway function disturbances (from predicted to hyperreactivity and obstruction) was also performed. The improvement in airways function was noticed only in 6 patient (20% studied). In patients with mitral valve disease airway function disturbances as obstruction and bronchial hyperreactivity persist in long term follow-up after mitral valve replacement. PMID- 11928658 TI - [Effects of spirometric screening in the community on smoking cessation]. AB - In the years 1998-2000 in the Centre of COPD and Respiratory Failure in Bydgoszcz a group of tobacco smokers at risk of COPD (over 40 years of age, with smoking history of more than 10 packyears) were studied. Every patient filled-in a simple questionnaire on clinical signs of COPD and tobacco habit, had a spirometry performed according to ATS standards and received a short antitobacco counseling together with a booklet on how to quit smoking. Out of 1072 patients studied in 1998, airway obstruction was found in 200. Eighty seven of these were current smokers (studied group-S). Another 90 current smokers with normal spirometry served as a control group-C. Both groups of smokers were invited in 1999 and 2000 to perform spirometry. Changes in smoking habit were recorded at each yearly visit. After one year 13 patients (15%) from the S group and 4 (4.5%) from the C group permanently stopped smoking. In 2000 24 patients (28%) in S group and 13 (14.7%) in C group permanently quit smoking. CONCLUSION: Spirometric screening in smokers at risk of COPD together with a minimal antismoking intervention seems to be a promising method of smoking cessation. PMID- 11928659 TI - [Clinical value of polymesam recording in diagnosis of obstructive sleep apnea. Can we replace polysomnography with polymesam?]. AB - Polymesam (PM) recordings was performed in 320 patients admitted to Sleep Laboratory with suspicion of OSA. OSA was diagnosed in 179 of them (55.9%), group (PM-Ch). These patients were obese (BMI--34.3 +/- 6.7 kg/m2) and had moderately severe OSA (RDI--41.5 +/- 19.9 and ODI--43.7 +/- 21.5). They suffered from excessive daytime sleepiness (ESS = 12.2 +/- 5.5). PM was negative in 141 person (44.1%), (PM-Z). Subjects PM-Z had significantly lower BMI and rarely suffered from excessive daytime sleepiness. In 38 subjects PM-Z a full PSG was performed. In 12 PSG confirmed OSA (AHI--31.6 +/- 19.9). Both studies (PM and PSG) were negative in 26 subjects. In 10 obese subjects PM-Ch full PSG confirmed diagnosis. CONCLUSIONS: PM recording can replace full PSG in majority of patients suspected of OSA. Patients with typical symptoms of OSA and negative PM require PSG. PMID- 11928660 TI - [Ten years experience of the sleep laboratory at the Institute of Tuberculosis and Lung Disease in Warsaw]. AB - Between 1991-2000 2052 patients (81% men and 19% women) were referred to our Sleep Laboratory because of OSA suspision. In 1194 (58%) subjects (88% men and 12% women) diagnosis of obstructive sleep apnoea (OSA, AHI > 10) was confirmed. In 430 of them (36%) mild OSA (AHI 11-25), in 243 (20%) moderate OSA (AHI 26-40), and in 521 (44%) severe OSA (AHI > 40) was diagnosed. Epworth sleepiness scale score in those groups was 10.4, 10.5 and 13.0 points respectively. 908 (76%) of patients with OSA were submitted to nCPAP treatment. Effective CPAP pressure ranged from 5 to 20 milibars, mean 8.4 mbars. In 21 patients upper airway resistance syndrome (UARS) was diagnosed. Central sleep apnoea, most frequently of Cheyne-Stokes respiration type was diagnosed in 13 patients. The most common diseases accompanying OSA were: systemic hypertension (46%), coronary heart disease (29%), diabetes (12%), and COPD (9%). Majority of OSA patients (61%) were obese (BMI > 30 kg/m2), 32% were over weight (BMI 25-30 kg/m2). Only 7% had normal body weight (BMI 20-25 kg/m2). Long-term (more than one year) compliance to treatment was found in 70% of patients prescribed CPAP. PMID- 11928661 TI - [Predictors of persistence of remission of the respiratory symptoms in school children]. AB - The evolution of the respiratory symptoms in school children hampers the interpretation of epidemiologic findings concerning potential risk factors. It is thus important to identify prognostic factors that predict symptoms' persistence or remission. A seven-year follow-up of 663 children showed that half of them experience the remission of cough, chest wheeze and attacks of dyspnea. The results of multivariate analysis showed that the persistence of cough is more likely to occur in children with a history of spastic bronchitis (logOR = 3.1; 95% CI: 1.3-7.1) and the presence of coal-stove in kitchen (logOR = 3.1: 95% CI: 1.4-6.9). The probability of persistence of wheeze depends on history of bronchitis (logOR = 3.9; 95% CI: 1.4-11.4), history of parental asthma(logOR = 4.8; 95% CI: 1.1-25.2) and diagnosis of pollen sensitivity (logOR = 3.9; 95% CI = 1.2-12.9). On the other hand the persistence of attack of dyspnea was associated with a shortness of breath on exertion(logOR = 4.7; 95% CI: 1.5-16.8). The results of the study confirm an important role of medical history in the assessment of prognosis of the respiratory health status in children. PMID- 11928662 TI - [Selected parameters of the oxidative-antioxidative balance in children with chronic severe bronchial asthma]. AB - The aim of this study was to evaluate the involvement of selected parameters of oxidative-antioxidative system in children with severe bronchial asthma. In our study the intensity of peroxide lipid oxidation was calculated by the contents of malonic dialdehyde in plasma. At the same time antioxidant activity was evaluated. The plasma contents of vitamin E and total antioxidant status were measured, as were activity of glutathione peroxidase and superoxide dismutase in blood. Our study was performed on 24 children treated for severe bronchial asthma. The control group consisted of 27 healthy children. The study has revealed that children suffering from bronchial asthma exhibit disturbances in the oxidative/antioxidative balance, manifested by increased oxidative activity and by changes in certain parameters of the antioxidative system. Our results support the theory of free radicals involvement in pathophysiology of severe bronchial asthma. PMID- 11928663 TI - [Tuberculous peritonitis as still probable cause of ascites]. AB - A 38-year-old man was admitted to hospital because of ascites, fever, weakness, abdominal pain. Chest x-ray revealed old post-tuberculous opacities in the left subclavicular region and fluid in left pleural cave. Cirrhosis of the liver and malignant disease were suspected. Treatment with antibiotics and evacuation of pleural fluid caused short-lasting improvement. After 6 weeks of diagnostic procedures parenchymal infiltrations in left lung were confirmed. In sputum tuberculous bacilli were found. Tuberculous peritonitis was confirmed histopathologically. Antituberculous treatment was successful. PMID- 11928664 TI - [Acromegaly and sleep apnoea syndrome--case report]. AB - We studied 65-year old, obese man suspected of obstructive sleep apnoea. He gave a history of loud snoring and excessive daytime sleepiness. We confirmed sleep apnoea syndrome during limited polysomnography with Polymesam (RDI--45/h, ODI- 47/h). Patient had mainly obstructive episodes, however central and mixed apnoeas constituted about 1/3 of all episodes. During hospitalization we observed exacerbation of coronary artery disease and diagnosed diabetes. Patient's coarsened facial features, macroglossia and large hands led us to suspect acromegaly. Brain MR study revealed small pituitary adenoma. Plasma GH and IGF-1 concentrations were increased. Active acromegaly was diagnosed and was proposed a surgical treatment but he refused. Symptoms of sleep apnoea relieved after CPAP treatment. After one year patient's condition remained stable. PMID- 11928666 TI - [Usefulness of the eosinophil count in induced sputum of asthmatic patients]. AB - Chronic mucosal inflammation is referred to as the primary cause of asthma. Treatment in the first place aims to prevent and reverse the above disease. It is possible to measure the inflammation relatively noninvasively and reliably by making use of induced sputum cell counts. The differential cell count points to the existence and sort of the inflammation (eosinophilic or neutrophilic) and the total cell count--to the intensity. Sputum eosinophilia responds to treatment with corticosteroid, while there is increasing evidence that an isolated neutrophilia does not. Clinical judgement of airway inflammation is further complicated due to the various types of inflammation and their inconsistent correlation with the clinical features. Hence, reliable measurement of induced sputum cell counts may be useful to guide treatment in clinical practice. Therefore, it should be considered how to make it more available. PMID- 11928665 TI - [Diagnosis and treatment of exercise-induced asthma]. PMID- 11928667 TI - [Antioxidative features of ambroxol--usefulness in COPD]. PMID- 11928668 TI - [Exposure to chemical carcinogens in Poland in 1999]. AB - Occupational exposure to chemical carcinogens in Poland, based on the data provided by the Central Register of Carcinogenic Agents, is presented. More than 220,000 employees in 2376 enterprises were exposed to chemical substances, mixtures and technological processes recognized as carcinogenic. The majority of them were present in the three regions (voivodships) of Poland: slaskim, dolnoslaskim and malopolskim. PMID- 11928669 TI - [Occupational exposure to chemicals in the manufacture of rubber tires]. AB - The work environment of the rubber industry company, producing various types of tires, was assessed and the workers of the plant were included in a cohort study of mortality. Concentrations of twenty chemical substances at 137 workposts were measured by employees of the plant laboratory and the sanitary and epidemiological station in 1981-1996. The mean values and concentration ranges were determined by departments and workposts. The excess of threshold limit values was analyzed. The workposts with exposure to agents possibly carcinogenic to humans were identified. The analysis performed will render it possible to calculate doses of cumulative exposure to given compounds among workers covered by the epidemiological study. PMID- 11928671 TI - [Assessment of exposure to bioaerosols in workplace ambient air during municipal waste collection and disposal]. AB - The assessment of the exposure to bioaerosols among workers engaged in the collection and disposal of municipal waste is presented. The workers were divided into the following groups, depending on the job performed: waste collectors (loaders, drivers), composting plant workers (heavy equipment operators, waste site workers), sorters (heavy equipment operators, waste sorters) and landfill site workers (heavy equipment operators, weighers). Air samples were also collected on the city streets and in flats. They were reference points to the results obtained. Air samples were collected at the breathing zone. The concentrations of dust, viable microorganisms (mesophilic and thermophilic bacteria, and fungi), endotoxins and the total number of microbiol cells were determined. The highest, individual exposure to dust was found in composting plant workers and waste collectors. The values exceeded considerably maximum allowable concentrations (by ten times at composting plant). The mean values for the above listed groups also exceeded the concentration of 4 mg/m3. Dust concentrations at other workposts were substantially lower. The concentrations in the city streets and in flats maintained at the level of 0.1 mg/m3. Endotoxin concentration at a protecting upper airway inflammation level (10 ng/m3) was exceeded in the majority (above 50%) of samples. That is why the average concentration in individual groups was also higher than 10 ng/m3. Street and flat samples included endotoxins at the level of 1 ng/m3. Taking as a criterion the obligatory total dust and endotoxin MAC value of 10 ng/m3 for assessing work hygiene conditions, it should be stated that waste collectors and composting plant workers were employed in conditions of poor hygiene. PMID- 11928670 TI - [Health risk among workers employed in rubber footwear plant]. AB - The aim of the study was to assess the health risk of workers performing specific jobs in the process of the rubber footwear production by defining the cause and length of temporary work disability, as well as mortality causes and level. The analysis was carried out in the groups of workers performing the following jobs: mixing, mill operation, pressing and vulcanizing (A); semi-product preparation and calendaring (B); finishing and sorting (C); production of polyvinyl chloride footwear (D); and auxiliary works (E). The sickness absence study covered all workers (208 men and 315 women) employed in a large rubber footwear company and performing all above-listed jobs in 1995. Standardized sick days ratio was used to analyze the risk of temporary work disability. Mortality rate was estimated on the basis of the results of the cohort study performed in the same company among workers who had worked at least three months during the years 1945-1985. The follow-up continued until 31 December 1997. The present study included sub cohorts composed of 5628 men and 7197 women, performing jobs listed above. The results of both studies indicated the enhanced risk of cardiovascular diseases among workers employed in the basic phases of the production process. The increased risk of the diseases of the digestive system was observed in men and women employed in: finishing, sorting and packing of the products (group C); in men involved in mixing, pressing and vulcanizing (group A); and in women engaged in auxiliary works (group E). In addition, the enhanced risk of sickness absence due to the diseases of the respiratory, digestive, or genitourinary systems was related to the enhanced risk of death from malignant neoplasms in a given site. The analysis showed that the temporary work disability may be regarded as a parameter useful in early assessment of health effects of the work environmental hazards. PMID- 11928672 TI - [Occupational exposure to gases emitted in mild and stainless steel welding]. AB - The objective of this work was to select optimal methods for determination of toxic gases (NOx, NO2, CO, CO2, O3) and to evaluate occupational exposure of welders to those gases. The survey covered workers employed in shipyards, and other metal product fabrication plants engaged in welding mild and stainless steel by different techniques (manual metal are, metal active gas, tungsten inert gas welding; gas, plasma, laser cutting and resistance welding). Personal and stationary air samples were collected to determine time weighted average (TWA) and short-term concentrations of gases. For determination of nitrogen oxides the following analytical techniques were employed: spectrophotometry with collection on liquid and solid sorbents and ion chromatography with collection on solid sorbents. All the gases were determined also by automatic or direct reading methods: flow or diffusion detector tubes and photometric and electrochemical analyzers. The determined TWA concentrations were below respective Maximum Allowable Concentrations (MAC) but exposure limits for short term exposure were exceeded in some cases. The average NO2 i NOx ratio was 1:4. According to Polish regulations regarding the MAC value for nitrogen oxides the analytical method should enable determination of total NOx by either direct or indirect simultaneous determination of both NO and NO2. The applicability of the spectrophotometric method of analysis of atmospheric NOx to determination of low NOx concentrations at welders working posts has been confirmed. PMID- 11928673 TI - [Dust exposure and cancer risk associated with amphibolite mining and processing]. AB - Mining and processing of amphibolite is associated with workers' exposure to dust containing asbestos minerals (actinolite, tremolite) and with the presence of respirable fibers, i.e. small particles above 5 microns long and below 3 microns in diameter (with length-to-diameter ratio higher than 3:1). Results of epidemiological and laboratory studies show that such dust may be responsible for the development of cancer in dust-exposed people. This work reports the measurement results of concentrations of total dust, respirable fibers and mineral composition of samples collected in plant mining and processing amphibolite rock. Based on the results, cumulated exposure was calculated for the 10-, 20- and 30-year exposure periods. The cumulated exposure was classified into two categories: 0.1-1.0 f/cm3 years and 1.0-10 f/cm3. x years. It has been found that mining and processing of amphibolite is associated with increased risk of death from mesothelioma--11.2 x 10(-5) (crushers--10 years of exposure) to 240.0 x 10(-5) (miners--30 years of exposure). The risk of excessive mortality from lung cancer was not high (below 1. x 10(-4)) for all workplaces and periods of exposure. PMID- 11928674 TI - [New sources of occupational exposure in modified or new technological processes]. AB - Mixtures of organic solvents with frequently changing composition are employed in many industries. It is true that dangerous solvents are eliminated from the work environment, but simultaneously new manufacturing techniques are implemented. Glues, varnishes and stains, containing chemicals of unknown toxicity and non recognized effects on the human body are extensively used in the footwear and furniture industries. The aim of our study was to identify new sources of occupational exposure. The tests were performed in plants of selected industries. Samples of glues, varnishes and wood stains were collected to glass vials (10 ml). The air-tight vials were kept at the temperature of +2 degrees C to +8 degrees C until removed for the analysis. After incubation period (T = 24 degrees C; t = 1 h), 100 microliters of gas phase was removed with gas-tight syringe. Organic solvents were identified by gas chromatography with mass spectrometry detection (GC/MSD). The following classes of organic compounds were detected in glues, varnishes and stains under study: alkanes, cycloalkanes, alcohols, ketones, esters, aromatic hydrocarbons and their numerous alkyl derivatives. The tests confirmed that glues, varnishes and stains contained chemicals with poorly recognized or totally unknown toxicity and toxicokinetics; MAC values were set for only few of them. PMID- 11928675 TI - [Sense of personal control and the level of occupational stress and related effects]. AB - There is opinion that the sense of personal control at workplace is an important factor determining the occupational stress and related effects. The group of 160 policemen were studied to verify empirically the above-mentioned opinion. The subjects assessed their level of sense of personal control, occupational stress, burnout and mental health. Statistical analysis revealed following significant correlation coefficients between the sense of personal control and stress (r = 0.33); job satisfaction (r = -0.44); emotional exhaustion (r = -0.28); depersonalization (r = -0.26); and feeling of personal accomplishment (r = 0.25); total score in GHQ-28 (r = -0.16). The results of the study let us draw a practical conclusion. In order to diminish the level of job stress it is required to introduce proper organizational changes enhancing the sense of personal control among workers. PMID- 11928676 TI - [How to solve the problem of smoking at workplaces]. AB - This paper addresses the problem of the cessation of tobacco smoking at workplaces in major enterprises of Poland. The data were collected in 2000 by means of a questionnaire sent to 755 enterprises employing at least 100 workers. The Polish law on the health protection against harmful effects of tobacco consumption forbids smoking at workplace premises except for those isolated and specially set-up for smoking. Almost 80% of enterprises introduced appropriate bans on smoking. 14% of which are total bans. However, only 40% of companies control whether the regulations are followed by employees and 75% of the firms set up smoking-rooms that could effectively isolate tobacco smoke. Over 60% of enterprises introduced additional procedures to convince their staff to quit smoking. Educational materials stressing the relationship between ill health and smoking are usually distributed. About 17% of enterprises have introduced disciplinary measures. Financial bonuses, psychological or pharmacological therapy are rather rare (around 1%). The main reasons for antismoking measures are: legal regulations and care of employee's health (60% each), and fire protection (50%). Economical aspects have rarely been reported (25%). Together with non-smokers, the enterprise management have been recognized as the most substantial champions of quitting smoking. About 85% of medical staff attribute this role to themselves, but only 15% of managerial workers perceive them as partners in antismoking campaign. In conclusion, there is a great need to encourage medical staff, managers and workers to become more active in implementing programs on health promotion at work with special emphasis put on antismoking campaigns. PMID- 11928677 TI - [Assessment of low frequency noise annoyance in steering premises according to subjective annoyance rating by workers. A pilot study]. AB - The study covered 192 workers aged 26-62 years (mean 41 +/- 6), employed at workposts in the control rooms. A questionnaire was applied as a main tool of the study. The aim of the study was subjective annoyance rating of low frequency noise at workplaces on a 100-degree rating scale. Using the same scale, the study subjects rated annoyance of reference wide-band noise (60 +/- 2, 70 +/- 2, 75 +/- 2 and 80 +/- 2 dB(A)) and reference low frequency noise (65 +/- 2, 70 +/- 2, 75 +/- 2 and 80 +/- 2 dB(A)) reproduced by phones. Questionnaire inquires were preceded by the measurements of A-, C- and G-weighted equivalent continuous sound pressure levels (LA eq.T, LC eq,T i LG eq,T). Noise with prominent low frequency components (10-250 Hz) at A-weighted sound pressure level of 47.7-65.2 dB was assessed as more than somewhat annoying (more than 25 degrees on the 100-degree annoyance rating scale) by 70% of workers examined. No correlation between subjective annoyance rating of the noise at the workplaces and LA eq,T or LG eq,T was noted. Statistically significant correlation (r = 0.23) between subjective annoyance rating and LC eq,T was observed. The subjective rating of annoyance of the reproduced low frequency noise and wide-band noise differed significantly. Low frequency noise was rated as more annoying than wide-band noise at the same A weighted sound pressure levels. PMID- 11928678 TI - [Beryllium--underestimated occupational health hazard in Poland]. AB - Beryllium is a metal used in different branches of industry. Inhalation is the most common way of exposure. Beryllium shows a high affinity to the lung tissue. It may act as toxic, carcinogenic and immunological agent. Chronic beryllium disease (CDB) is very difficult to diagnose because of its similarity to sarcoidosis. The lymphocyte proliferation assay is a very helpful test in CBD diagnosis. It seems reasonable to include this test into the body of the standard diagnostic procedures in workers exposed to beryllium. Early detection of hypersensitivity to beryllium, before CBD symptoms occur will allow to minimize adverse health effects. PMID- 11928679 TI - [Nervous system disorders induced by occupational exposure to aluminium compounds: a literature review]. AB - This is a review of the literature on the effect of aluminum (Al) and its compounds on the nervous system. The role of aluminum in etiology of some degenerative diseases of the nervous system, e.g. Alzheimer disease, amyotrophic lateral sclerosis or dementia, is presented. The special attention was turned to the effects of aluminum on the nervous system functions in persons occupationally exposed to metal-containing dusts and fumes, manifested mostly by neurobehavioral disorders and changes in the brain bioelectric functions and less frequently pronounced by clinical neurological symptoms. PMID- 11928680 TI - [Occupational risk factors for breast cancer in the epidemiological studies]. AB - The paper reviews data on the occupational risk factors, which role in the etiology of breast cancer in women has been analyzed and estimated in the epidemiological studies. The results of the studies are presented and discussed. An increased incidence or risk of death from breast cancer was observed in some studies among teachers, nurses and physicians, managerial occupations, beauticians and hairdressers, chemists, telephone and telegraph operators, workers of the pharmaceutical and chemical industry. Some epidemiological studies indicate that breast cancer incidence might be related to occupational exposure to electromagnetic fields, ionizing radiation, low physical activity, organic solvents, selected pesticides. However, owing to inconsistency of the study results and methodological weaknesses (imprecise classification of exposure, low response rate, uncontrolled confounding) no definite conclusion can be drawn. PMID- 11928681 TI - National Comprehensive Cancer Network 6th Annual Conference Proceedings. March 1 4, 2001. PMID- 11928682 TI - Sentinel lymph node evaluation. PMID- 11928684 TI - Transcatheter cardiovascular therapeutics 2000. PMID- 11928685 TI - [Catastrophic antiphospholipid syndrome revealed by a cerebrovascular accident: a case report]. AB - We reported the case of a patient with catastrophic antiphospholipid syndrome revealed by an ischemic stroke, and who presented within six weeks several visceral failures, with among others surrenal, cardiac, splenic, cutaneous and hematologic complications. PMID- 11928686 TI - [Orthodontic anomalies in children with open-mouth posture]. AB - Open-mouth posture is related to the development of orthodontic anomalies. The etiology of these anomalies is discussed. Guidelines for diagnosis, prevention and interceptive measures are given. PMID- 11928687 TI - [Oral pathology in children. Part I. Diseases of the oral mucosa]. AB - An overview is present of diseases of the oral mucosa that are more or less limited to occurrence in children or that are more difficult to recognize in children. An example of the latter is geographic tongue. PMID- 11928688 TI - [Atypical pain in the upper jaw]. PMID- 11928689 TI - Letting history repeat itself. PMID- 11928690 TI - A patient's view. PMID- 11928691 TI - Genetic programming over context-free languages with linear constraints for the knapsack problem: first results. AB - In this paper, we introduce genetic programming over context-free languages with linear constraints for combinatorial optimization, apply this method to several variants of the multidimensional knapsack problem, and discuss its performance relative to Michalewicz's genetic algorithm with penalty functions. With respect to Michalewicz's approach, we demonstrate that genetic programming over context free languages with linear constraints improves convergence. A final result is that genetic programming over context-free languages with linear constraints is ideally suited to modeling complementarities between items in a knapsack problem: The more complementarities in the problem, the stronger the performance in comparison to its competitors. PMID- 11928692 TI - Adenocarcinoma of the lung in a 32-year-old man: is it worthwhile to check for lung cancer in young adults? PMID- 11928693 TI - Conjunctival involvement in a patient with cutaneous malignant melanoma. PMID- 11928694 TI - Prophylactic cranial irradiation in limited-disease small-cell lung cancer - why, when, how much? PMID- 11928695 TI - Does the extent of lymphadenectomy have impact on the prognosis of patients with pancreatic cancer? PMID- 11928696 TI - Minimal access surgery for cancer treatment. PMID- 11928697 TI - Is there a standard for the palliative treatment of melanoma? PMID- 11928698 TI - Misconceptions about cryosurgery in warts of the nail unit. PMID- 11928699 TI - Regarding treatment of myxoid cysts. PMID- 11928700 TI - A new adulterated silicone epidemic: Java, Indonesia. PMID- 11928701 TI - Use of split-thickness graft to preserve the undersurface and width of airborne postauricular pedicle flaps. PMID- 11928702 TI - [Hypercalcemia and impaired renal function in an 74 year-old woman]. PMID- 11928703 TI - [A blue man and odd hematological findings]. PMID- 11928704 TI - Characteristics of sex-biased dispersal and gene flow in coastal river otters: implications for natural recolonization of extirpated populations. AB - River otters (Lontra canadensis) were extirpated from much of their historic distribution because of exposure to pollution and urbanization, resulting in expansive reintroduction programmes that continue today for this and other species of otters worldwide. Bioaccumulation of toxins negatively affects fecundity among mustelids, but high vagility and different dispersal distances between genders may permit otter populations to recover from extirpation caused by localized environmental pollution. Without understanding the influence of factors such as social structure and sex-biased dispersal on genetic variation and gene flow among populations, effects of local extirpation and the potential for natural recolonization (i.e. the need for translocations) cannot be assessed. We studied gene flow among seven study areas for river otters (n = 110 otters) inhabiting marine environments in Prince William Sound, Alaska, USA. Using nine DNA microsatellite markers and assignment tests, we calculated immigration rates and dispersal distances and tested for isolation by distance. In addition, we radiotracked 55 individuals in three areas to determine characteristics of dispersal. Gender differences in sociality and spatial relationships resulted in different dispersal distances. Male river otters had greater gene flow among close populations (within 16-30 km) mostly via breeding dispersal, but both genders exhibited an equal, low probability of natal dispersal; and some females dispersed 60-90 km. These data, obtained in a coastal environment without anthropogenic barriers to dispersal (e.g. habitat fragmentation or urbanization), may serve as baseline data for predicting dispersal under optimal conditions. Our data may indicate that natural recolonization of coastal river otters following local extirpation could be a slow process because of low dispersal among females, and recolonization may be substantially delayed unless viable populations occurred nearby. Because of significant isolation by distance for male otters and low gene flow for females, translocations should be undertaken with caution to help preserve genetic diversity in this species. PMID- 11928705 TI - Spatial autocorrelation and linkage of Mendelian RAPD markers in a population of Picea abies Karst. AB - The spatial clustering of single- and di-locus genotypes in a natural, continuous population of Norway spruce was investigated using 69 Mendelian Random Amplified Polymorphic DNA (RAPD) markers that covered about 15 of the species genome, and whose linkage relationships were known. Spatial autocorrelation techniques and randomization tests, applied to both single- and di-locus genotypes, revealed a weak, though significant, spatial structure at the scale 0-200 m (5 of single locus and 7 of di-locus genotypes). To assess the relative importance of isolation by distance and linkage between markers on their spatial genetic structuring, we grouped joins between sampled trees into equivalence categories expected to show similar, specific patterns of spatial distribution under isolation by distance. Results from both single- and di-locus analyses were consistent with the existence of patches of like homozygotes (about 8 and 11 of loci at the single- and di-locus level, respectively) surrounded by a mix of like heterozygotes. Similar structuring has been predicted by simulation models under isolation by distance and selective neutrality. Overall, linkage between markers accounted for an increase of spatial clumping of di-locus genotypes involving tightly linked loci with recombination fractions up to 0.1, a consequence of limited, stochastic spread of single-locus genotypes in space. Our results support the hypothesis that isolation by distance and linkage have a small, though significant, effect even within continuous forest tree populations. In general, the spatial distribution of multilocus genotypes within populations should be interpreted with caution when linkage relationships among the markers used are unknown. PMID- 11928706 TI - Gene flow, dispersal, and nested clade analysis among populations of the stonefly Peltoperla tarteri in the southern Appalachians. AB - We examined the genetic structure and phylogeography of populations of the stonefly Peltoperla tarteri in the Southern Appalachians to determine the extent and likely mechanism for dispersal of this stream insect. A 454-base-pair (bp) portion of the mitochondrial control region was sequenced from a minimum of 20 individuals from eight populations. Pairwise FST and exact tests showed high levels of differentiation among almost all populations except those on the same stream. amova analysis detected significant genetic differentiation between streams within drainages (phi(SD) = 0.14, P < 0.001), and there was a slight positive correlation between aquatic distance and genetic distance (r = 0.295, P = 0.03). According to nested clade analysis, the present day pattern of genetic variation in P. tarteri is the result of a historical range expansion coupled with restricted gene flow with isolation by distance. Together, these analyses suggest that adult dispersal is limited and that movement by larvae is the primary dispersal mechanism for P. tarteri. PMID- 11928707 TI - Comparative population structure and genetic diversity of Arceuthobium americanum (Viscaceae) and its Pinus host species: insight into host-parasite evolution in parasitic angiosperms. AB - In a recent study we revealed that the parasitic angiosperm Arceuthobium americanum is comprised of three distinct genetic races, each associated with a different host in regions of allopatry. In order to assess the role of host identity and geographical isolation on race formation in A. americanum, we compared the genetic population structure of this parasite with that of its three principal hosts, Pinus banksiana, Pinus contorta var. latifolia and Pinus contorta var. murrayana. Despite the fact that A. americanum was divided into three genetic races, hosts were divided into only two genetic groups: (i) Pinus banksiana and hybrids, and (ii) P. contorta var. latifolia and var. murrayana. These findings suggest that factors such as geographical isolation and adaptation to different environmental conditions are important for race formation in the absence of host-driven selection pressures. To assess factors impacting population structure at the fine-scale, genetic and geographical distance matrices of host and parasite were compared within A. americanum races. The lack of a relationship between genetic and geographical distance matrices suggests that isolation-by-distance plays a negligible role at this level. The effect of geographical isolation may have been diminished because of the influence of factors such as random seed dispersal by animal vectors or adaptation to nongeographically patterned environmental conditions. Host-parasite interactions might also have impacted the fine-scale structure of A. americanum because the parasite and host were found to have similar patterns of gene flow. PMID- 11928708 TI - Microsatellite DNA and recent statistical methods in wildlife conservation management: applications in Alpine ibex [Capra ibex(ibex)]. AB - We evaluated the usefulness of microsatellites and recently developed statistical methods for the conservation management of fragmented and reintroduced populations, using the alpine ibex (Capra ibex) as a model species. First, we assessed the effects of past reintroduction programmes on genetic diversity and population differentiation considering different population sizes and histories. We show that genetic variability in ibex populations (HE 0.13) is among the lowest reported from microsatellites in mammal species, and that the Alpi Marittime-Mercantour population has suffered from a severe genetic bottleneck associated with its reintroduction. Second, using a computer-simulation approach, we provide examples and rough guidelines for translocation programmes concerning the number and origin of individuals for future reintroductions and for the reinforcement of populations with low genetic variability. Finally, we use the ibex microsatellite data to assess the usefulness of several published statistical tests for detecting population bottlenecks and assigning individuals to their population of origin. This study illustrates that microsatellites allow: (i) evaluation of alternative translocation scenarios by simulating different numbers and origins of migrants; (ii) identification of bottlenecked populations (especially using the Wilcoxon signed-ranks test); and (iii) population assignment with a high certainty (P < 0.001) of almost 100 of the individuals (or trophies or carcasses) from two distant populations (especially using stucture or whichrun software). PMID- 11928709 TI - Community analysis of arbuscular mycorrhizal fungi associated with Ammophila arenaria in Dutch coastal sand dunes. AB - A polymerase chain reaction-denaturing gradient gel electrophoresis (PCR-DGGE) approach for the detection and characterization of arbuscular mycorrhizal fungi (AMF) 18S ribosomal DNA (rDNA) was developed and applied to the study of AMF communities associated with the main sand-stabilizing plant species of the Dutch sand dunes, marram grass (Ammophila arenaria, L.). DNA was extracted directly from plant roots, soil or isolated AMF spores, and prominent bands resulting from AMF-specific DGGE profiles were excised for sequence analysis. This strategy provided a robust means of detecting and identifying AMF-like species without the use of trap plant cultivation methods. A number of Glomus-like and Scutellospora like sequences was detected, including a putatively novel Glomus species, and differences were observed in the dominant AMF-like populations detected in healthy vs. degenerating stands of A. arenaria and in bulk sand dune soil. It has previously been suggested that plant pathogens, such as fungi and nematodes, may contribute to the decline of A. arenaria. Although no causal relationship can be drawn between the observed differences in the dominantly detected AMF-like populations and the vitality of plant growth, these results indicate that mutualistic interactions between this plant and AMF should not be overlooked when examining the role of soil-borne microorganisms in vegetation dynamics. In addition, there were discrepancies observed between the AMF-like groups detected in spore populations vs. direct 18S rDNA analysis of root material, corroborating previous suggestions that spore inspection alone may poorly represent actual AMF population structure. PMID- 11928710 TI - Optimizing strategies for the detection of gene-nutrient interactions in the aetiology of colorectal neoplasia: discussion and conclusions from the ECP consensus-forming workshop. Brussels, 25-26 September 2000. PMID- 11928711 TI - Fundamental role of heme oxygenase in the protection against ischemic acute renal failure. AB - Oxidative stress conditions such as oxidant stimuli, inflammation, exposure to xenobiotics and ionizing irradiation provoke cellular responses, principally involving transcriptional activation of genes encoding proteins that participate in the defense against oxidative tissue injuries. Excess of free heme, which is released from hemeproteins under these conditions, may constitute a major threat because it catalyzes the formation of reactive oxygen species. Exposure of mammalian cells to oxidative stimuli induces heme oxygenase-1 (HO-1), the rate limiting enzyme in heme degradation, as well as the 32-kDa heat shock protein. In various tissue injury systems, HO-1 induction has been shown to confer protection, while its abrogation has been shown to accelerate cellular injuries. In this review, recent findings concerning the role of HO-1 as a protective response against oxidative stress conditions are summarized, with a particular emphasis on its protective role in ischemic acute renal failure. PMID- 11928712 TI - Neuronal nicotinic receptor and psychiatric disorders: functional and behavioral effects of nicotine. AB - Both retrospective and prospective clinical studies have demonstrated positive associations of smoking with psychiatric disorders such as schizophrenia, depression and anxiety. Neuronal nicotinic acetylcholine receptors (nAChR) belong to a family of ligand-gated ion channels that are widely distributed in the brain. The pre-synaptically located nAChR, which are composed of alpha3 or alpha4 subunits in combination with beta2 subunit on axon terminals, modulate the multiple transmission release. Several studies indicated which individual nicotinic receptor subtype is responsible for mediating each of the behavioral effects of nicotine. A reduced number of alpha7 nicotinic receptor subtypes in the hippocampus were reported in schizophrenic patients. In addition, it was assumed that nicotine provided useful therapeutic treatment for a variety of cognitive impairments including those found in Alzheimer's disease, schizophrenia and attention deficit hyperactive disorder. Both alpha7 and alpha4beta2 nicotinic receptors in the hippocampus are involved in these phenomena. In the genetic depressive rats, nicotine showed antidepressant-like effects in forced swim models of depression, suggesting the involvement of alpha4beta2 nicotinic receptor in this phenomenon. Thus, it appears likely that pre-synaptic nAChR on monoaminergic fibers are composed of alpha3 or alpha4 subunits in combination with the beta2 subunit, and these subunit compositions mediate dopaminergic and noradrenergic release, and glutamate is mainly controlled by the alpha7 subunit. All these findings suggest that nicotine and other nicotinic drugs warrant further study for possible clinical prescription to psychiatric disorders. PMID- 11928713 TI - Suppression of endotoxin-induced renal tumor necrosis factor-alpha and interleukin-6 mRNA by renin-angiotensin system inhibitors. AB - The present study was designed to clarify the role of angiotensin II (Ang II) in modulating renal tumor necrosis factor (TNF)-alpha and interleukin-6 (IL-6) production and to investigate the effect of one dose of Ang II inhibitor on cytokines production following lipopolysaccharide (LPS) to cause endotoxemia. Two studies were performed: 1) Ang II was infused intravenously at a rate of 0.2 microg/kg per minute for 4 h in rats and then kidneys were collected to assay TNF alpha and IL-6 mRNA levels; 2) Four-week-old Wistar rats pre-treated with angiotensin-converting enzyme inhibitor, enalapril, or type I Ang II-receptor antagonist, TCV-116, were injected with LPS (0.1, 0.5, 1.0 mg, i.p.), and then 2 or 4 h later, kidneys were collected to assay TNF-alpha, IL-6, renin and angiotensinogen mRNA levels. After a 4-h intravenous infusion of Ang II, renal TNF-alpha or IL-6 mRNA level significantly increased 1.9-fold or 2.1-fold (each P<0.05) to the control level, respectively. LPS stimulated TNF-alpha, IL-6 and angiotensinogen mRNA levels in the kidney but in rats given enalapril or TCV-116, LPS-induced IL-6 and TNF-alpha mRNA levels were completely suppressed (each P<0.05). This suggests that a single dose of renin-angiotensin system inhibitor suppressed renal IL-6 and TNF-alpha production and may prevent cytokine-induced renal damage during endotoxemia. PMID- 11928714 TI - Existence of functional alpha1A- and alpha1D- but no alpha1B-adrenoceptor subtypes in rat common carotid arteries. AB - Using the cannula inserting method, vasoconstrictor responses to alpha1 adrenoceptor agonists (noradrenaline [NA], phenylephrine [PE] and methoxamine [ME]) and effects of alpha1-adrenoceptor antagonists (WB4101, chloroethylclonidine [CEC] and BMY7378) were investigated in isolated and perfused rat common carotid arteries. The rank order of agonist potency and efficacy was NA = PE > ME. Either WB4101 or BYM7378 inhibited NA- and PE-induced constrictions in a dose-related manner. CEC did not inhibit the NA- and PE induced responses. The ME-induced responses were also significantly blocked by either WB4101 or BMY7378. From these results, it is concluded that there are functional alpha1A- and alpha1D-adrenoceptor subtypes in rat common carotid arteries, but no functional alpha1B subtype. PMID- 11928715 TI - Involvement of calmodulin inhibition in analgesia induced with low doses of intrathecal trifluoperazine. AB - We examined which of the known properties of trifluoperazine, including calmodulin inhibition, are involved in its analgesic effect. Furthermore, we tried to find any possible interaction between opioidergic system and calmodulin inhibition-induced analgesia. Intrathecal trifluoperazine (1, 10, 100 microg) showed a biphasic effect in the formalin test; i.e., analgesia at relatively low doses (1, 10 microg) and hyperalgesia at a high dose (100 microg). No analgesic effects were observed after intrathecal injection of sulpiride (1, 10, 100 microg), atropine (0.1, 1, 10 microg), phentolamine (0.1, 1, 10 microg) and brompheniramine (0.1, 1, 10 microg). Meanwhile, intrathecal calmidazolium (10, 50, 250 microg) induced a dose-dependent analgesia. Histamine (1 microg), physostigmine (1 microg), bromocriptine (1 microg) and norepinephrine (1 microg) did not affect trifluoperazine-induced analgesia. Calcium (20 microg) attenuated the antinociceptive effect of trifluoperazine and inhibited the analgesic effect of calmidazolium. Finally, naloxone (2 mg/kg) decreased trifluoperazine-induced antinociception but did not have any effects on calmidazolium-induced analgesia. We concluded that calmodulin inhibition may be involved in the analgesia produced by trifluoperazine. With increasing doses of trifluoperazine, the algesic effect seems to overcome the analgesic effect. It is also suggested that the opioidergic system does not interact with calmodulin inhibition-induced analgesia even though this system has a possible role in trifluoperazine-induced analgesia. PMID- 11928716 TI - Novel mutations in C-terminal channel region of the ryanodine receptor in malignant hyperthermia patients. AB - Malignant hyperthermia (MH) is a pharmacogenetical complication of general anesthesia resulting from abnormal Ca2+-induced Ca2+ release (CICR) via the type 1 ryanodine receptor (RyR1) in skeletal muscles. In this study, we analyzed the genomic DNAs prepared for determination of all the 106 exons of the RyR1 gene from blood samples donated by two MH patients with extremely high CICR rates in their biopsied skeletal muscles and a clear history of MH incidence. Two novel point mutations were found in the exons 96 and 101 with alterations in the coded amino acids within the C-terminal channel region, i.e., Pro4668 to Ser and Leu4838 to Val. The latter mutation was found in both MH patients. Rabbit RyR1 channels carrying corresponding mutations were expressed in CHO cells for functional assay. It was found that the L to V but not the P to S mutation of the RyR1 resulted in enhanced Ca2+ release activity. These results indicate that the L4838V mutation is responsible for the MH incidence. The L4838V mutation is unique because it is the mutation first found within a hydrophobic transmembrane segment of the channel region and should provide further information on the function of the RyR1 as well as for genetic diagnosis of MH. PMID- 11928718 TI - Novel monoamine oxidase inhibitors, 3-(2-aminoethoxy)-1,2-benzisoxazole derivatives, and their differential reversibility. AB - Although possible usefulness of non-selective monoamine oxidase (MAO) inhibitors for Parkinson's disease therapy has been suggested in the literature, MAO inhibitors whose inhibition is reversible and have dual action to both MAO-A and B subtypes is not available yet. Subtype selectivity and reversibility of a series of novel MAO inhibitors, 3-(2-aminoethoxy)-1,2-benzisoxazole derivatives, were studied. Several dual MAO inhibitors, which inhibit both MAO-A and -B, were obtained. When administered to mice, their effects were generally reversible. Among the derivatives, RS-1636 and RS-1653 had much longer duration of brain MAO B inhibition than that of MAO-A. In vitro, the inhibited MAO-A activity by these compounds was partially recovered by buffer change at 4 degrees C, while little MAO-B activity was recovered. Although it is not fully elucidated yet, the reversibility of these inhibitors is probably determined primarily by this dissociation profile. This unique differential reversibility indicates that optimization of the balance of actions can be achieved by differentiating reversibility to each target molecule. PMID- 11928717 TI - Augmented natriuretic peptide-induced guanylyl cyclase activity and vasodilation in experimental hyperglycemic rats. AB - The present study was aimed to investigate whether hyperglycemia may alter the regulation of vascular natriuretic peptide receptors (NPR). The hyperglycemia was induced in rats by the treatment with streptozotocin (50 mg/kg, i.v.). The expression of different subtypes of NPR was determined in the thoracic aorta by reverse transcriptase-polymerase chain reaction and quantitative in vitro receptor autoradiography. The isometric tension and the guanylyl cyclase activity of the isolated thoracic aorta in response to natriuretic peptides were also determined. Following the treatment with streptozotocin, the plasma concentration of atrial natriuretic peptide (ANP) was significantly increased. The expression of NPR-A was increased, while that of NPR-C was reduced. The receptor binding study demonstrated an increased maximal binding capacity of NPR, with its affinity not significantly altered. The magnitude of vasodilation and guanylyl cyclase activity in response to ANP was significantly increased. On the other hand, the vasodilator response as well as the tissue formation of cGMP in response to acetylcholine or sodium nitroprusside was significantly reduced. These results indicate that the hyperglycemia may cause an altered regulation of vascular NPR. PMID- 11928719 TI - Beneficial effects of angiotensin-converting enzyme inhibition in adriamycin induced cardiomyopathy in hamsters. AB - This study was performed to determine whether angiotensin (Ang) II-forming enzymes, angiotensin converting enzyme (ACE) and chymase might contribute to the development of adriamycin-induced cardiomyopathy in hamsters. Hamsters were administered adriamycin (2.0 mg/kg per day, i.p.) three times weekly for 2 weeks. In the ACE inhibitor-treated group, the hamsters received lisinopril (20 mg/kg per day, p.o.) for 2 weeks after the last injection of adriamycin. The 4-week mortality rates of the vehicle- and ACE inhibitor-treated hamsters were 44% and 12%, respectively. In comparison to the age-matched hamsters used as the control hamsters, a significant decrease in cardiac function and a significant increase in the ratio of the heart weight to the body weight were observed in the vehicle hamsters. Cardiac ACE activity, but not the chymase activity, in the vehicle hamsters was significantly increased in comparison to that in the control hamsters. In the ACE inhibitor-treated group, the increased ACE activity was reduced significantly, and the cardiac hypertrophy and dysfunction were improved significantly. In adriamycin-induced cardiomyopathic hamsters, cardiac ACE activity was increased and ACE inhibition significantly improved cardiac function and survival rate, indicating that cardiac ACE, but not the chymase, plays the pivotal role in the development of the adriamycin-induced cardiomyopathy. PMID- 11928720 TI - Endothelium-dependent relaxation of rat mesenteric arterial rings by a Phoneutria nigriventer venom fraction. AB - Phoneutria nigriventer spider venom has been described as acting on several cardiovascular sites. In the present paper, a semi-purified fraction of this spider venom was studied to observe any contractile or relaxing effect in rat mesenteric arterial rings (MAR). Spider venom was first fractionated by gel filtration and subsequently by gradual isocratic steps in 0.1% trifluoroacetic acid. The first fraction of this last fractionation step is studied in the present paper and due to its main effect, it was named NORF (nitric oxide releasing fraction). No direct contractile effect was induced by NORF in relaxed MAR, suggesting no NORF-induced neurotransmitter release in this preparation. No significant influence of NORF was observed on concentration-response curves to phenylephrine on endothelium-denuded MAR, but a significant inhibitory shift of concentration-respense curves was observed on endothelium-preserved MAR (EC50 = 0.39 +/- 0.07 microM for control and EC50 = 0.68 +/- 0.14 microM with NORF). NORF induced concentration-dependent relaxation in endothelium-preserved phenylephrine pre-contracted MAR but not in endothelium-denuded MAR. NORF-induced relaxation was inhibited by the nitric oxide synthase inhibitor L-NAME (N(omega)-nitro arginine methyl ester). Indomethacin or HOE-140 (D-Arg-[Hyp3,Thi5,D-Tic7,Oic8] bradykinin) had no significant effect on NORF-induced relaxation. Acetylcholine- and NORF-induced relaxation of pre-contracted MAR were differently inhibited by atropine. The pA2 value for atropine-acetylcholine was 9.78 +/- 0.06 and that for atropine-NORF was 8.53 +/- 0.30 (P<0.01). These observations suggest that NORF induces concentration-dependent liberation of nitric oxide from MAR endothelium and that a non-muscarinic mechanism might be involved in this effect. Our data suggest no involvement of prostanoids or bradykinin in the relaxing mechanism. PMID- 11928721 TI - Involvement of proteasome in endothelin-1 production in cultured vascular endothelial cells. AB - We examined whether the proteasome could regulate endothelin (ET)-1 production in vascular endothelial cells (ECs). A proteasome inhibitor N-benzyloxycarbonyl-Ile Glu (O-t-Bu)-Ala-leucinal (PSI) significantly decreased ET-1 release from ECs by about 25% of the basal release. PSI also suppressed tumor necrosis factor (TNF) alpha-induced ET-1 release from ECs in a dose-dependent manner. Similar inhibitory effects were observed using another proteasome inhibitor lactacystin, whereas a calpain inhibitor calpeptin had no apparent effect on ET-1 release. Furthermore, PSI significantly attenuated prepro ET-1 mRNA expression under basal and TNF-alpha-stimulated conditions. Electrophoretic mobility shift assay showed that proteasome inhibitors diminished TNF-alpha-stimulated nuclear factor-kappa B (NF-kappaB) activation in ECs. Pretreatment with antioxidants, pyrrolidine dithiocarbamate and alpha-lipoic acid, both of which are known to be suppressors of NF-kappaB activation, effectively attenuated basal and TNF-alpha-induced ET-1 release. Thus, a proteasome-dependent proteolytic pathway is at least partly involved in ET-1 production under basal conditions, and this proteolytic pathway seems to have a crucial role in ET-1 production enhanced by TNF-alpha. The reduction of NF-kappaB activation may be involved in the mechanisms for suppressive effects of proteasome inhibitors on ET-1 gene transcription and the consequent decrease in ET-1 mRNA expression and ET-1 release. PMID- 11928723 TI - Antiproteinuric effect of KD3-671, an angiotensin II type 1 receptor antagonist, in rats with accelerated passive Heymann nephritis. AB - The antiproteinuric effect of KD3-671 (2-propyl-8-oxo-1-[(2'-(H-tetrazole-5 yl)biphenyl-4-yl)methyl]-4,5,6,7-tetrahydrocycloheptimidazole), an angiotensin II type 1 receptor antagonist, was compared with that of enalapril, an angiotensin 11-converting enzyme inhibitor, using an experimental model of membranous nephropathy. KD3-671 (3, 10 and 30 mg/kg per day) and enalapril (30 mg/kg per day) were given p.o. for 40 days, respectively. KD3-671 (30 mg/kg per day) inhibited the elevation of proteinuria and plasma total cholesterol. On the other hand, enalapril showed only a tendency to diminish these parameters. KD3-671 had an antiproteinuric effect in rats with accelerated passive Heymann nephritis. These findings provide considerable encouragement for the clinical development of KD3-671. PMID- 11928722 TI - Effects of T-82, a new quinoline derivative, on cholinesterase activity and extracellular acetylcholine concentration in rat brain. AB - The effects of T-82 (2-[2-(1-benzylpiperidin-4-yl)ethyl]-2,3-dihydro-9-methoxy-1H pyrrolo [3,4-b]quinolin-1-one hemifumarate), a new quinoline derivative, on acetylcholinesterase (AChE) activity and acetylcholine (ACh) release were compared with those of the well-known cholinesterase inhibitors tacrine and E2020. T-82, tacrine and E2020 all concentration-dependently inhibited AChE in rat brain homogenate (IC50 = 109.4, 84.2 and 11.8 nM, respectively). In addition, although tacrine strongly inhibited butyrylcholinesterase (BuChE), T-82 and E2020 showed only weak activity on BuChE in human plasma. In ex vivo experiments, intraperitoneal administration of T-82 at a dose of 30 mg/kg inhibited AChE activity in the hippocampus, frontal cortex and parietal cortex of rats. The effect of T-82 on the extracellular ACh concentration in rat brain was measured using in vivo microdialysis. T-82 at doses of 10 and 30 mg/kg, i.p. increased the extracellular ACh concentration in the hippocampus and striatum in a dose dependent manner. These findings suggest that T-82 activates the central cholinergic system by selectively inhibiting AChE activity, while weakly affecting peripheral BuChE activity, and that T-82 increases the extracellular ACh concentration in the brain, which is followed by inhibited AChE activity. PMID- 11928724 TI - Abatement of morphine-induced slowing in gastrointestinal transit by Dai-kenchu to, a traditional Japanese herbal medicine. AB - As a way of alleviating severe constipation in cancer patients taking morphine to relieve pain, effects of Dai-kenchu-to (DKT), a traditional Japanese herbal medicine (Kampo medicine), on gastrointestinal transit in mice or on the isolated guinea pig ileum were studied in special reference to morphine. Without altering the anti-nociceptive effect of morphine, DKT was significantly effective against morphine-induced disorder of gastrointestinal transit in mice as assessed by the charcoal meal test for the intestine and measurement of transit time for the colon tract. The results of in vitro studies with guinea pig ileum suggest that abatement of morphine-induced disorder of transit by DKT is caused by both moderate contraction of morphine-treated longitudinal muscle and relaxation of morphine-induced tonic contraction of circular muscle. PMID- 11928725 TI - Glutamate exacerbates amyloid beta1-42-induced impairment of long-term potentiation in rat hippocampal slices. AB - Amyloid beta (A beta) is the principal constituent of senile plaques in Alzheimer's disease patients. We investigated whether A beta and glutamate affect long-term potentiation (LTP) in rat hippocampal slices. Pretreatment with 1 microM A beta1-42 alone for 3 h slightly inhibited LTP; however, the potentiation was maintained for 60 min. Although the impairment was not observed by pretreatment with 30 microM glutamate alone for 3 h, pretreatment with A beta1-42 and glutamate impaired LTP significantly. These results raise the possibility that neurotoxicity of A beta is exacerbated by the enhancement of susceptibility to excitatory amino acids. PMID- 11928727 TI - Unifocal amyloidosis: a rare cause of spinal cord compression. AB - A 75-year-old man presented with a 3-month history of progressive paraparesis due to an extradural mass causing cord compression at the T7 level of the thoracic spine. He underwent decompressive surgery, and later vertebrectomy and cage fixation. Histologically, the lesion was a localized mass of amyloid associated with a clonal plasma cell infiltrate. Localized 'amyloidomas' of the spine are rare, evolve slowly and often have a good prognosis following surgery. PMID- 11928728 TI - The working time directive: the potential impact on a neurosurgical centre's medical manpower and service delivery from 2004. PMID- 11928726 TI - Early recurrence of benign meningioma correlates with expression of mini chromosome maintenance-2 protein. AB - We have investigated the potential utility of monoclonal antibodies against mini chromosome maintenance-2 protein (Mcm2) in predicting meningioma recurrence. MCM proteins are members of the DNA-binding prereplicative complex and are essential for eukaryotic DNA replication. They are present throughout the cell cycle, but are down-regulated in quiescence and cell differentiation, making them specific markers of proliferating cells. We analysed 10 benign meningiomas that subsequently recurred within a 5-year period, together with 20 matched non recurrent benign meningiomas. There was no significant correlation between histological subtype, mitotic count or Ki-67 labelling index and tumour recurrence. We observed that whilst the average Mcm2 labelling index (LI) of the tumour section as a whole (LI(Ave)) is not significantly different between recurrent and nonrecurrent meningiomas, the Mcm2 labelling index in the area of highest proliferative activity within the tumour section (LI(Max)) is significantly higher in recurrent meningiomas (p < 0.0001). Seven out of the 10 recurrent meningiomas displayed a Mcm2 LI((Max) greater than 30%, compared to 0 out of 20 for non-recurrent tumours. In conclusion, these results suggest that analysis of Mcm2 expression may facilitate identification of patients with a high risk of meningioma recurrence, for whom adjuvant radiotherapy may be of benefit. PMID- 11928729 TI - Nephrotic syndrome: more than just oedema. AB - Oedema is the commonest presenting symptom and sign in nephrotic syndrome. Hypercholesterolaemia, thromboembolic events, and infectious complications may also be features. Three patients are described, each of whose nephrotic syndrome presented with a less common symptom or sign--recurrent pleural effusion, hypercholesterolaemia and oedema, pulmonary embolism--and, as a result, experienced some diagnostic delay. By forgetting to consider nephrotic syndrome, and its underlying causes, there may be inappropriate investigations and treatment for the patient. PMID- 11928730 TI - Diabetic ketoacidosis in acromegaly: indication for long-term insulin therapy? PMID- 11928731 TI - Eosinophilia associated with olanzapine. PMID- 11928732 TI - Report from the Eighth Conference on Retroviruses and Opportunistic Infections. PMID- 11928733 TI - Summaries for patients. Using magnetic resonance direct thrombus imaging to diagnose deep-vein thrombosis in the lower legs. PMID- 11928734 TI - Summaries for patients. Initial evaluation of rectal bleeding in young persons: a cost-effectiveness analysis. PMID- 11928735 TI - Summaries for patients. Comparing the quality of diabetes care by generalists and specialists. PMID- 11928736 TI - Summaries for patients. Reporting on statistical methods to adjust for confounding. PMID- 11928737 TI - Summaries for patients. Aspirin for the prevention of heart attacks in people without previous cardiovascular events: recommendations from the United States Preventive Services Task Force. PMID- 11928738 TI - Summaries for patients. Alcohol and risk for heart failure. PMID- 11928739 TI - Summaries for patients. Drinking less alcohol improves heart function in people with alcohol-related heart failure. PMID- 11928740 TI - Summaries for patients. Dietary patterns and the risk for type 2 diabetes in U.S. men. PMID- 11928741 TI - Summaries for patients. Delayed-onset heparin-induced thrombocytopenia. PMID- 11928742 TI - Physicians and capital punishment. PMID- 11928743 TI - Pharmacist scope of practice. PMID- 11928744 TI - Measuring quality. PMID- 11928745 TI - Caring for patients at the end of life. PMID- 11928748 TI - Bush's advisers on ethics discuss human cloning. PMID- 11928746 TI - Proxy consent to organ donation by incompetents. PMID- 11928749 TI - Viewing-zone enlargement method for sampled hologram that uses high-order diffraction. AB - We demonstrate a method of enlarging the viewing zone for holography that has holograms with a pixel structure. First, aliasing generated by the sampling of a hologram by pixel is described. Next the high-order diffracted beams reproduced from the hologram that contains aliasing are explained. Finally, we show that the viewing zone can be enlarged by combining these high-order reconstructed beams from the hologram with aliasing. PMID- 11928750 TI - Color image recognition by use of a joint transform correlator of three liquid crystal televisions. AB - We present a joint transform correlator for color image recognition by using three liquid-crystal spatial light modulators. A method for simultaneously obtaining the correlation peaks of red, green, and blue is proposed and experimentally demonstrated. PMID- 11928751 TI - Convergent optical correlator aligment based on frequency filtering. AB - The convergent correlator is widely used but it presents the drawback of the alignment requirements: fine focusing of the input scene Fourier transform on the filter plane, filter centering, scaling the scene Fourier transform to match the filter size, and azimuth matching of the filter with the input scene. We propose a set of tests to obtain a precise alignment of the convergent correlator. These methods are based on frequency filtering properties and they are applicable either for amplitude input or for phase-encoded input. The tests we present allow us to fulfill all the alignment requirements. The theory on which these tests are based is explained. The experimental results obtained during the alignment procedure are presented. We show some additional verifications of the correct alignment of the convergent correlator. PMID- 11928752 TI - Quadratic cost functional for wave-front reconstruction. AB - A quadratic cost functional for reconstruction of a high-resolution wave front from a coarse wave front is presented. The functional uses as data the position and the direction of the coarse wave front that had previously been computed with a ray-tracing method. This functional uses an optical relationship between the ray information and the wave front's shape to reconstruct a high-density wave front. The performance of the proposed functional is illustrated by reconstruction of complicated wave fronts for which this functional has an accuracy that is superior to that of conventional interpolation methods. PMID- 11928753 TI - Holographic optical elements recorded in silver halide sensitized gelatin emulsions. Part 2. Reflection holographic optical elements. AB - Silver halide sensitized gelatin (SHSG) holograms are similar to holograms recorded in dichromated gelatin (DCG), the main recording material for holographic optical elements (HOEs). The drawback of DCG is its low energetic sensitivity and limited spectral response. Silver halide materials can be processed in such away that the final hologram will have properties like a DCG hologram. Recently this technique has become more interesting since the introduction of new ultra-fine-grain silver halide (AgHal) emulsions. In particular, high spatial-frequency fringes associated with HOEs of the reflection type are difficult to construct when SHSG processing methods are employed. Therefore an optimized processing technique for reflection HOEs recorded in the new AgHal materials is introduced. Diffraction efficiencies over 90% can be obtained repeatably for reflection diffraction gratings. Understanding the importance of a selective hardening process has made it possible to obtain results similar to conventional DCG processing. The main advantage of the SHSG process is that high-sensitivity recording can be performed with laser wavelengths anywhere within the visible spectrum. This simplifies the manufacturing of high-quality, large-format HOEs, also including high-quality display holograms of the reflection type in both monochrome and full color. PMID- 11928754 TI - Behavior of the point-spread function in photon-limited confocal microscopy. AB - We study the behavior of the point-spread function (PSF) of the confocal scanning optical microscope (CSOM) when the available optical energy density from the sample plane is low (<7.5 microJoule/micrometers2). The PSF profile is analyzed under three photon-limited imaging conditions: (1) reflection-type CSOM with a weak source and a perfectly reflecting sample, (2) reflection-type CSOM with a strong illumination source and a weak sample, and (3) fluorescence CSOM with a weak fluorescent sample. Linfoot's image quality criteria of fidelity, structural content, and correlation quality are used to assess the reproducibility of the PSF profile as a function of the photon number. Low photon numbers yield a PSF profile that is difficult to maintain from one location in the sample plane to another. The optical sectioning capability of the CSOM was found to deteriorate more quickly against light power reduction than its transverse resolving power. The signal-to-noise ratio of the scanned CSOM image improves exponentially with the photon number from the sample plane. The noise that is generated by an unstable PSF has an average amplitude that decreases exponentially with the photon number and is significant only at low photon numbers. The CSOM image quality deteriorates because of spurious high-frequency components, degradation in the PSF dynamic range, and varying resolving power. PMID- 11928755 TI - Signed-negabinary-arithmetic-based optical computing by use of a single liquid crystal-display panel. AB - Based on the negabinary number representation, parallel one-step arithmetic operations (that is, addition and subtraction), logical operations, and matrix vector multiplication on data have been optically implemented, by use of a two dimensional spatial-encoding technique. For addition and subtraction, one of the operands in decimal form is converted into the unsigned negabinary form, whereas the other decimal number is represented in the signed negabinary form. The result of operation is obtained in the mixed negabinary form and is converted back into decimal. Matrix-vector multiplication for unsigned negabinary numbers is achieved through the convolution technique. Both of the operands for logical operation are converted to their signed negabinary forms. All operations are implemented by use of a unique optical architecture. The use of a single liquid-crystal-display panel to spatially encode the input data, operational kernels, and decoding masks have simplified the architecture as well as reduced the cost and complexity. PMID- 11928756 TI - Intensity, polarization, and phase information in optical disk systems. AB - Digital information in optical data storage systems can be encoded in the intensity, in the polarization state, or in the phase of a carrier laser beam. Intensity modulation is achieved at the surface of the storage medium either through destructive interference from surface-relief features (e.g., CD or DVD pits) or through reflectivity variations (e.g., alteration of optical constants of phase-change media). Magneto-optical materials make use of the polar magneto optical Kerr effect to produce polarization modulations of the focused beam reflected from the storage medium. Both surface-relief structures and material property variations can create, at the exit pupil of the objective lens of the optical pickup, a phase modulation (this, in addition to any intensity or polarization modulation or both). Current optical data storage systems do not make use of this phase information, whose recovery could potentially increase the strength of the readout signal. We show how all three mechanisms can be exploited in a scanning optical microscope to reconstruct the recorded (or embedded) data patterns on various types of optical disk. PMID- 11928757 TI - Analytic performance analysis based on material properties for electroabsorptive asymmetric Fabry-Perot reflection modulators. AB - Simple and accurate analytic expressions are provided for the maximum reflectivity and tolerances of an impedance-matched asymmetric Fabry-Perot used as a high-contrast spatial light modulator when electroabsorptive quantum wells provide loss modulation. When the device geometry is optimized, these expressions depend only on material properties. The maximum reflectivity depends only on the fractional absorption change and is independent of the front-mirror reflectivity. The most important tolerance is on the flatness of crystal growth; the fractional length tolerance is proportional to the absorption coefficient. These formulas agree with experimentally reported results from multiple-quantum-well modulators and previous numerical analyses; they are useful for quickly predicting optimized performance of possible new materials. The normally on and normally off geometries are compared. The effect of finite back-mirror reflectivity is clarified. Deviations from impedance match enable increased reflectance difference at the expense of contrast ratio, an approach which is evaluated as a function of material parameters. PMID- 11928758 TI - Role of bile acid measurement in pregnancy. AB - The diagnosis of obstetric cholestasis (OC) has serious implications for maternal, and especially fetal, health. Total serum bile acid concentration is an important investigation in any woman with itching in the absence of a rash during pregnancy. Results should be available within 1-2 working days. Pruritus plus raised total bile acids in the third trimester suggests a diagnosis of OC. Other biochemical abnormalities, usually a raised alanine aminotransferase, accompany or follow the finding of raised bile acids. Bile acids are sensitive, but not specific, markers for OC. The diagnosis is one of exclusion. The clinician should remain alert to other causes of liver dysfunction, either specific to pregnancy or not. Effective treatment is available that improves maternal biochemical indices and well-being. It is not clear whether such treatment reduces the risks to the fetus. More active management of OC pregnancies has reduced the associated fetal and perinatal mortality. PMID- 11928759 TI - Stains, labels and detection strategies for nucleic acids assays. AB - Selected developments and trends in stains, labels and strategies for detecting and measuring nucleic acids (DNA, RNA) and related molecules [e.g. oligo(deoxy)nucleotides, nucleic acid fragments and polymerase chain reaction products] are surveyed based on the literature in the final decade of the 20th century (1991-2000). During this period, important families of cyanine dyes were developed for sensitive detection of double-stranded DNA, single-stranded DNA, and oligo(deoxy)nucleotides in gels and in solution, and families of energy transfer primers were produced for DNA sequencing applications. The continuing quest for improved labels for hybridization assays has produced a series of candidate labels including genes encoding enzymes, microparticles (e.g. quantum dots, nanocrystals, phosphors), and new examples of the fluorophore (e.g. cyanine dyes) and enzyme class of labels (e.g. firefly luciferase mutants). Label detection technologies for use in northern and southern blotting assays have focused on luminescent methods, particularly enhanced chemiluminescence for peroxidase labels and adamantyl 1,2-dioxetanes for alkaline phosphatase labels. Sets of labels have been selected to meet the demands of multicolour assays (e.g. four-colour sequencing and single nucleotide primer extension assays). Non separation assay formats have emerged based on fluorescence polarization, fluorescence energy transfer (TaqMan, molecular beacons) and channelling principles. Microanalytical devices (microchips), high-throughput simultaneous test arrays (microarrays, gene chips), capillary electrophoretic analysis and dipstick devices have presented new challenges and requirements for nucleic acid detection, and fluorescent methods currently dominate in many of these applications. PMID- 11928760 TI - Biochemical markers of neonatal sepsis: value of procalcitonin in the emergency setting. AB - BACKGROUND: We evaluated procalcitonin (PCT) assay in the emergency diagnosis of neonatal bacterial infection, especially in preterm infants, relative to C reactive protein (CRP) and fibrinogen. METHODS: One hundred and twenty neonates (32 preterm), of whom 21 were infected, were tested. RESULTS: Concentrations of PCT, CRP and fibrinogen in uninfected infants were not affected by gestational age at birth. Concentrations of CRP and PCT increased rapidly during the first 24 h of life, while fibrinogen concentrations increased gradually from birth. All marker concentrations were significantly greater in neonates with bacterial infection. Receiver-operating characterstic analysis showed that optimum cut-off values for fibrinogen, CRP and PCT were 3.0 g/L, 7.5 mg/L and 2.5 microg/L respectively, for the diagnosis of sepsis at birth. CONCLUSIONS: Determination of PCT is of value in excluding bacterial infection in neonates since it has a negative predictive value of 93%. PMID- 11928762 TI - Optimizing the availability of 'stat' laboratory tests using Shewhart 'C' control charts. AB - BACKGROUND: We describe a general strategy for optimizing the availability of 'stat' out-of-hours laboratory tests to the particular clinical needs of health care institutions. METHODS: We initially introduced a consensus menu of 'stat' tests and prospectively monitored for 5 years all additional requests for 'stat' clinical biochemistry tests in a metropolitan tertiary referral university teaching hospital. Laboratory staff triaged 'stat' requests, and clinical biochemistry consultants reviewed requests not already performed routinely. The frequency of these requests was monitored on a Shewhart 'c' control chart. A quality system certified to ISO 9001 was used to assure laboratory compliance with procedures. Various interventions were tested using the Shewhart 'c' control chart to monitor their effectiveness. RESULTS: Matching the timing of analytical assays with the time of sample collection had no significant effect on the number of 'stat' requests. Implementation of a hospital-wide laboratory information system also had no significant effect on the number of 'stat' requests. The most effective strategy consisted of optimization of the test menu to match request patterns, combined with the introduction of a laboratory quality system. CONCLUSIONS: Within our institution, this strategy resulted in a sevenfold reduction in 'stat' requests, from one per 2,200 specimens to fewer than one per 32,000 specimens. PMID- 11928761 TI - Diurnal variation in lipoprotein lipase activity. AB - BACKGROUND: We investigated whether variations in lipoprotein lipase activity, a key post-prandial enzyme involved in the removal of circulating dietary triglycerides, could contribute to the previously described nocturnal lipid intolerance. METHODS: We studied lipoprotein lipase activity in 12 healthy volunteers (five women, seven men) at 11:30 h and 23:30 h on two separate occasions. Subjects consumed a high-fat mixed meal at 07:30 h for the morning study or 19:30 h for the evening study. Then, after a 4-h fast, subjects were given an intravenous bolus of 7,500 U heparin. Blood samples were collected before and 15 min after heparin administration for measurement of lipoprotein lipase, hepatic lipase, triglycerides and non-esterified fatty acids concentrations. RESULTS: Post-prandial post-heparin lipoprotein lipase activity was greater in the morning than in the evening (16.5 +/- 1.4 versus 14.4 +/- 1.0 micromol oleate/mL/h; P< 0.05). Post-prandial post-heparin hepatic lipase activity was also greater in the morning than in the evening (8.7 +/- 1.5 versus 81 +/- 1.6 micromol oleate/mL/h; P= 0.002). There were no other significant diurnal differences. CONCLUSION: We report a diurnal variation in post-prandial lipoprotein lipase activity. This is consistent with the notion that decreased nocturnal insulin sensitivity extends to insulin's actions on lipoprotein lipase and provides a possible explanation for nocturnal lipid intolerance. PMID- 11928763 TI - Evaluation of kryptor cardiac troponin I immunoassay. AB - BACKGROUND AND METHODS: Cardiac troponin I (cTnl) is well recognized as a specific marker for myocardial infarction. A fully automated, random access, fluorescent immunoassay for cTnl was evaluated in comparison with an established assay (Stratus cTnl) using samples from healthy subjects and from patients with cardiac disease. RESULTS: The detection and precision were acceptable, and no interference was observed from bilirubin, triglycerides, haemoglobin, rheumatoid factor, drugs (aspirin, dopamine, digoxin) or heparin. Results on the Kryptor cTnl assay correlated with those on the Stratus II cTnI assay. The optimum Kryptor cTnl concentration for acute myocardial infarction was 0.63 microg/L, with a sensitivity of 89% [95% confidence interval (CI): 75.9-96.3] and specificity of 98.9% (95% CI: 93.9-99.8). CONCLUSION: Kryptor cTnl immunoassay is suitable for use in the diagnosis of myocardial infarction. PMID- 11928765 TI - Naturally occurring mutation, Asp70his, in human butyrylcholinesterase. AB - BACKGROUND: People with genetic variants of butyrylcholinesterase can have hours of prolonged apnoea after a normal dose of succinylcholine or mivacurium. METHODS: Serum samples from 308 persons living in mid-USA were phenotyped to identify the atypical and fluoride variants. 308 samples were analysed for the K variant by DNA amplification, digestion with Mae III and gel electrophoresis. Amplified DNA from 16 samples was sequenced to identify the D70G, T243M and D70H mutations. Values for kcat and Km were determined for the D70H mutant BChE expressed in 293T cells. RESULTS: A new mutation, Asp70His, was identified. This mutation is located in the peripheral anionic site of butyrylcholinesterase, where it causes a 10-fold decrease in binding affinity for positively charged substrates. CONCLUSION: People homozygous for the Asp70His mutation are expected to have prolonged apnoea in response to succinylcholine or mivacurium, similar to people with the Asp70Gly mutation. PMID- 11928764 TI - Interpretation of plasma brain natriuretic peptide concentrations may require adjustment for patient's age. AB - BACKGROUND: Brain natriuretic peptide (BNP) is increasingly recognized as a practical, reliable and accurate indicator of left ventricular function and may have an important prognostic role in determining outcome following myocardial infarction and heart failure. METHODS AND RESULTS: We studied a total of 60 patients referred for dobutamine-stress echocardiography to evaluate possible reversible myocardial ischaemia. There was no echocardiographic evidence of structural or functional heart disease in any of the patients included in the study. Blood was sampled for BNP at rest, prior to dobutamine-stress. We found a significant linear correlation between BNP concentrations and patients' age, in the absence of structural or functional heart disease (r(s) = 0.53, 95% CI 0.25 0.73, P<0.0001). CONCLUSION: Our observations suggest the need for consideration of patient's age when interpreting BNP concentrations in the population. PMID- 11928766 TI - Triploid pregnancy detected incidentally by Down's syndrome screening. AB - Maternal serum screening for Down's syndrome and trisomy 18 identifies pregnancies with a greater risk of these abnormalities, which are then followed up by karyotyping of cells collected either by amniocentesis or by chorionic villus sampling. These techniques complement ultrasonography, which gives accurate gestational dating as well as identifying structural abnormalities. Other chromosomal abnormalities are sometimes detected by virtue of atypical maternal screening results. This report illustrates a case of triploidy, a lethal abnormality, detected incidentally due to an exceptionally high human chorionic gonadotrophin result identified during Down's syndrome screening. This allowed appropriate counselling of the parents followed by a decision to terminate the pregnancy, avoiding the potential trauma of a spontaneous miscarriage or, if born live, death of the baby. Termination of the pregnancy also resolved associated maternal hyperthyroidism. PMID- 11928767 TI - Falsely increased prostate-specific antigen concentration attributed to heterophilic antibodies. AB - We describe a 50-year-old man with a serum total prostatic-specific antigen concentration of 67.6 microg/L (reference range 0-4 microg/L) revealed as part of an annual health check programme. Clinical examination failed to identify any prostatic abnormality, and laboratory investigation indicated positive interference due to heterophilic antibodies. PMID- 11928768 TI - Reporting of creatinine concentration. PMID- 11928769 TI - Limited clinical utility of high-sensitivity plasma C-reactive protein assays. AB - Many recent studies have shown a relationship between plasma C-reactive protein (CRP) concentrations and risk of cardiovascular disease. However, the intra individual variation in plasma CRP concentrations is large. Use of plasma CRP measurement in individual patients is likely to result in many being misclassified in their risk status. Use of repeated measurements is not a practical solution to this problem. PMID- 11928770 TI - Cystatin C. AB - Clinical biochemists have long known the analytical and clinical limitations of creatinine and creatinine clearance measurement in the assessment of glomerular filtration rate (GFR). This background is reviewed in the article before assessing the utility of cystatin C, the most promising replacement biochemical marker yet identified. Cystatin C has been used in clinical research studies for more than 20 years and yet has been introduced into clinical practice in very few centres, firstly in Lund in Sweden and now Carshalton in the UK. Why is this? The review compares our ability to measure creatinine and cystatin C and their relative sensitivity and specificity for changes in GFR. Comparison is made of cystatin C with creatinine as screening tests for early renal dysfunction and for monitoring its progression where issues of reference ranges and within-individual variation are important. The superiority of cystatin C as a screening test is probably accepted but there are still concerns about non-renal influences upon its circulating concentration, particularly steroid therapy, insufficient data on the influence of malignancy and a general lack of prospective clinical studies confirming the prognostic significance of cystatin C. PMID- 11928771 TI - Emotional intelligence and the good community. AB - Suggests various ways in which the concept of emotional intelligence (EI) can inform community research and action. Begins by defining EI and reviewing the growing body of research on its contribution to educational, health, and occupational outcomes. Next distinguishes between EI and the related concept of emotional and social competence. Then considers the connection between EI and several topics of interest to community action researchers including prevention and wellness promotion, implementation and dissemination of innovative programs, community development, and the value of diversity. Concludes by suggesting that the ecological perspective can help inform research and practice relating to EI. PMID- 11928773 TI - The 2001 Society for Community Research and Action Award for Distinguished Contributions to Theory and Research in Community Psychology: Rhona S. Weinstein. PMID- 11928772 TI - The process through which an advocacy intervention resulted in positive change for battered women over time. AB - A prior experimental evaluation of a community-based advocacy program for women with abusive partners demonstrated positive change in the lives of women even 2 years postintervention (C M. Sullivan & D. I. Bybee, 1999). The current study explored the complex mediational process through which this change occurred, using longitudinal structural equation modeling and formal tests of mediation. As hypothesized, the advocacy intervention first resulted in women successfully obtaining desired community resources and increasing their social support, which enhanced their overall quality of life. This improvement in well-being appeared to serve as a protective factor from subsequent abuse, as women who received the intervention were significantly less likely to be abused at 2-year follow-up compared with women in the control condition. Increased quality of life completely mediated the impact of the advocacy intervention on later reabuse. Discussion places advocacy for women in the context of other efforts that are needed to build an effective community response to preventing intimate violence against women. PMID- 11928774 TI - Evaluating attributions for an illness based upon the name: chronic fatigue syndrome, myalgic encephalopathy and Florence Nightingale disease. AB - In recent years, considerable discussion has occurred about stigma surrounding the name given to an illness currently known as chronic fatigue syndrome (CFS). Although patients and medical personnel have expressed varying opinions on this issue, no studies have evaluated how beliefs about the illness change based upon the type of name used for diagnostic purposes. Proposals have been put forth to rename the illness with an eponym (a famous patient's or researcher's name) or with a less trivial sounding, more medically based type of name. In this study, attributions about CFS were measured in three groups of medical trainees. All groups read the same case study of a person with classic symptoms of chronic fatigue syndrome, with the only difference being in the type of name given. Trainees then were asked to provide attributions about certain aspects of the illness, including its cause, severity, and prognosis. Results suggested that, across name conditions, most trainees appeared to consider the symptom complex of CFS a serious illness resulting in poor quality of life. In addition, findings indicated that the name, chronic fatigue syndrome, may be regarded less seriously than the Myalgic Encephalopathy name with respect to some important aspects of the illness. In this study, specialty of medical trainee also played a role in how the illness was perceived. PMID- 11928775 TI - Overcoming inequality in schooling: a call to action for community psychology. AB - Community psychology, indeed psychology as a discipline, has been largely absent from the table of school reform. Schools are critical socializing forces in society and serve as the one institution through which the full diversity of our child population passes. At the start of the 21st century, despite successive waves of legislation, the goals of the civil rights struggle for equality in educational opportunity have yet to be achieved. Negative self-fulfilling prophecies, reflected at individual, interpersonal, institutional, and societal levels, play a critical role in creating and perpetuating unequal opportunities to learn. Such effects as well as pathways for preventive intervention are best understood through ecological lenses. Our field must commit a greater share of resources to collaborative and systemic change for a broader learning so that all children, regardless of their differences, have continuing and nonstigmatized opportunities to develop into competent adults. PMID- 11928776 TI - The spirit of community psychology. 2001 Seymour B. Sarason Award Address. AB - Community psychology as a field may have lost some of its original spirit; a spirit dominant at the founding of the field. Spirit is a difficult concept to discuss since it is a concept beyond usual discourse in psychology as a denotative, measurable and verifiable concept. Taking the lead from William James, Spirit, however elusive, is most essential to a sense of self. I offer three suggestions to enhance and to make our individual and collective spirit more vital: (1) Continuing education in the history of community psychology; (2) Creating time to share stories about our work; (3) Creating safe settings to enlarge our spirit. These ideas are offered as pragmatic suggestions to enable us to create more coherence between our personal and professional selves. PMID- 11928777 TI - The contributions of Rudolf Moos. PMID- 11928778 TI - The mystery of human context and coping: an unraveling of clues. AB - Community and clinical psychology share a fundamental focus: to understand the interplay between human contexts, coping, and adaptation. To highlight recent progress in this area, I offer a guiding conceptual framework and discuss 8 propositions about environment and coping. The propositions consider such issues as patterns of social climate and coping and their links to personal development and dysfunction, the connections between ongoing life circumstances and intervention programs, the role of personal characteristics in matching individuals and environments, and the value of placing specific settings in an ecological context. I then focus on 8 enigmas, such as how to identify conceptually unifying dimensions of diverse social contexts, how to model the processes involved in person-environment transactions, how to understand the link between adversity and personal growth, how to examine the generality of models across ethnic and cultural groups, and how to enhance positive carryover from intervention programs to ongoing life contexts. I close by addressing some implications of these issues for a vision of a dynamic community psychology. PMID- 11928779 TI - Empowerment evaluation: building communities of practice and a culture of learning. AB - Empowerment evaluation is the use of evaluation concepts, techniques, and findings to foster improvement and self-determination. Program participants- including clients--conduct their own evaluations: an outside evaluator often serves as a coach or additional facilitator depending on internal program capabilities. Empowerment evaluation has three steps: 1) establishing a mission; 2) taking stock; and 3) planning for the future. These three steps build capacity. They also build a sense of community, often referred to as communities of practice. Empowerment evaluation also helps to create a culture of learning and evaluation within an organization or community. PMID- 11928780 TI - Comparative study of age, sex, illness profile, between suburban, rural and central urban population in general practice. AB - To investigate the differences between rural suburban and central urban patterns in General Practise, in terms of Patient age, sex and illness profiles, in Winter. Using a laptop computer, all relevant patient parameters were entered during the surgery consultation as each patient attended. The totals for each parameter were subsequently computed and using the Microsoft Excel program comparative pie charts for urban and rural categories were compiled, in percentage terms. SETTING: Cork. In the rural area 60% of those attending were male, while in the urban area only 43% were male. There were 2.75 times more people over 65 years old attending the urban surgery. However there were 80% more under 14 year olds and 43% more in the 15 to 64 year age group attending the rural surgery. PMID- 11928781 TI - Malignant otitis externa--a high index of suspicion is still needed for diagnosis. AB - Malignant otitis externa is a destructive inflammatory process of the petrous temporal bone which if untreated leads to osteomyelitis of the skull base and can be fatal. It is more common in immunocompromised and elderly insulin-dependant diabetic patients and is caused by infection with Pseudomonas species. Despite a range of laboratory and radiological tests it still remains difficult to diagnose, particularly in the early stages when it can be treated medically. We describe three cases which presented to this department in the past twelve months. In all cases the diagnosis was made clinically and confirmed per operatively. Interestingly all three cases were relatively young patients who did not have an immunocompromised status and were not diabetic. PMID- 11928782 TI - The effect of a change from conventional cardiac enzymes to troponin I on overall hospital costs in patients with suspected myocardial infarction. AB - A random sample of patients presenting to this hospital in 1996 and 2000 with chest pain was assessed retrospectively with respect to patient bed stay and associated costs. The laboratory testing protocol had been changed from traditional cardiac markers AST, CK and CKMB, to troponin I, in the intervening period. The average bed stay for patients with chest pain of non-AMI origin was reduced by 2 days, as a result of the change in testing protocol. As ward costs contribute 49% of total cost of treatment, this resulted in decreased cost per patient, and more efficient use of hospital beds. PMID- 11928783 TI - Treatment of psychotic depression associated with steroid therapy in Churg Strauss syndrome. AB - A case of depressive psychosis associated with corticosteroid therapy for the treatment of Churg-Strauss syndrome is described. We outline the patient's response to different antidepressant and antipsychotic medications in this case. A significant improvement with low doses of Venlafaxine is open for further discussion and study. PMID- 11928784 TI - Accidental ingestion of cotton bud stick during alcohol intoxication: an unusual cause of caecal perforation. AB - A first reported case of caecal perforation by an ingested blunt foreign body is described. This 31-year-old female did not relate a history of accidental swallowing of a cotton bud stick on admission. The cause of her right iliac fossa pain (i.e. caecal perforation by a cotton bud stick) became only apparent when she underwent operation for suspected appendicitis. A simple closure of this caecal perforation was undertaken. The patient made a complete recovery from this ordeal. Caecal perforation by an ingested blunt foreign body can occur after it has safely negotiated the ileocaecal valve. PMID- 11928785 TI - Compartment syndrome without pain! AB - We report the case of a young male patient who underwent intra-medullary nailing for a closed, displaced mid-shaft fracture of tibia and fibula. He was commenced on patient controlled analgesia post-operatively. A diagnosis of compartment syndrome in the patient's leg was delayed because he did not exhibit a pain response. This ultimately resulted in a below-knee amputation of the patient's leg. We caution against the use of patient controlled analgesia in any traumatised limb distal to the hip or the shoulder. PMID- 11928786 TI - Bupropion (Zyban) toxicity. AB - Bupropion is a monocyclic antidepressant structurally related to amphetamine. Zyban, a sustained-release formulation of bupropion hydrochloride, was recently released in Ireland, as a smoking cessation aid. In the initial 6 months since it's introduction, 12 overdose cases have been reported to The National Poisons Information Centre. 8 patients developed symptoms of toxicity. Common features included tachycardia, drowsiness, hallucinations and convulsions. Two patients developed severe cardiac arrhythmias, including one patient who was resuscitated following a cardiac arrest. All patients recovered without sequelae. We report a case of a 31 year old female who required admission to the Intensive Care Unit for ventilation and full supportive therapy, following ingestion of 13.5g bupropion. Recurrent seizures were treated with diazepam and broad complex tachycardia was successfully treated with adenosine. Zyban caused significant neurological and cardiovascular toxicity in overdose. The potential toxic effects should be considered when prescribing it as a smoking cessation aid. PMID- 11928787 TI - Synovial cyst of the hip joint: an unusual cause of a pulsating groin mass. AB - Synovial cysts of the hip joint occur most frequently in patients with rheumatoid arthritis (1). Despite the increasing frequency of arthroplasty of rheumatoid hips, case reports documenting synovial cysts remain extremely rare. This case report emphasizes the need for the surgeon to include a synovial cyst in the differential diagnosis of a painless groin mass, particularly in patients with rheumatoid arthritis. PMID- 11928788 TI - Patient use of the Internet and health strategy. PMID- 11928789 TI - Long-term headache following epidural analgesia. AB - Epidural analgesia is widely used in modern obstetric practice. Complications are uncommon but if they occur may result in significant morbidity. Post dural puncture headache (PDPH), in particular, may be particularly incapacitating especially if left untreated. In this report we describe the management of a case of unrecognised and undiagnosed PDPH. PMID- 11928790 TI - Medical screening--advice to doctors. PMID- 11928791 TI - Botulinum toxin--Jekyll or Hyde? PMID- 11928792 TI - Genetic hearing loss. PMID- 11928793 TI - Essential parameters in breast cancer: histopathology reporting guidelines. PMID- 11928794 TI - Randomized clinical trials in oncology commentary. AB - In this article the author addresses the major questions and problems that arrise while analyzing randomized clinical trials. A brief description is given of three problems, which are how to bring the derived information into broad applicability, when are the results of a trial significant enough to make it a standard for care and the issue of quality control. PMID- 11928795 TI - Randomized clinical trials in soft tissue sarcoma. AB - What has been learned thus far from the very best prospective randomized controlled clinical trials for soft tissue sarcoma that will guide physicians in the quest for evidence-based treatment decision making? Based on the evidence provided in the following articles, we have learned that limb-sparing surgery with adjuvant radiation is equivalent (and therefore preferable) to a radical amputation in controlling local disease. We also learned that adding radiation therapy (both external beam and brachytherapy) to a complete surgical resection of extremity soft tissue sarcoma significantly improves local control over surgery alone. This is true for high-grade lesions and may be true (external beam) for larger low-grade lesions also. Local control, however, does not govern overall survival as most patients who die from this disease do so from systemic (not local) manifestations. Finally, further studies are required to identify the role of adjuvant chemotherapy in the treatment of soft tissue sarcoma as no single study has demonstrated a consistent long-term survival benefit with the use of any chemotherapeutic regimen. Based on a recent large meta-analysis, however, if there is any benefit to adjuvant doxorubicin based chemotherapy, it must be a very small one. PMID- 11928796 TI - Randomized clinical trials in gastric cancer. AB - A total of 52 prospective, randomized controlled trials (RCT), published from 1975 to 2000, were reviewed for gastric cancer. The primary focus of these efforts has been the use of chemotherapy in patients with metastatic or locally advanced disease, accounting for 23 of the 52 trials. In comparison, there were only six surgical trials evaluating the extent of either primary resection or lymphadenectomy. PMID- 11928797 TI - Randomized clinical trials in colon cancer. AB - Prospective randomized trials involving the surgical and chemotherapeutic treatment of resectable colon cancer are reviewed. Emphasis is placed on those trials that have impacted the current management of primary colon cancer. PMID- 11928798 TI - Randomized clinical trials in rectal and anal cancer. AB - Treatment of rectal cancer has changed dramatically over the past two decades. Radical surgery alone for resectable rectal cancer has been replaced by combined modality therapy. Interest in optimizing sphincter preservation and quality of life motivated surgeons to seek less radical surgery without compromising oncologic principles. PMID- 11928799 TI - Randomized clinical trials in advanced and metastatic colorectal carcinoma. AB - The treatment of advanced colorectal cancer has long been a dilemma for clinicians. After many years of therapy limited to 5-fluorouracil (5-FU)-based chemotherapy, a number of new discoveries have significantly advanced this field in the last decade. The advent of irinotecan has given us hope for a second line chemotherapeutic agent and a possible new combination for first line chemotherapy. PMID- 11928800 TI - Randomized clinical trials in melanoma. AB - Efforts to improve the survival of patients with metastatic melanoma have led to 67 prospective randomized clinical trials investigating the use of agents comprising three main areas of therapy: systemic chemotherapy-based regimens, immunotherapy, and vaccine therapy PMID- 11928801 TI - Evidence-based oncology project. AB - This article gives a brief overview of what can be gained from randomized clinical trials and the problems that are faced when examining the results of these trials. It also discusses the two essential elements driving the evidence based oncology project, the first being the growth of MEDLINE and similar databases of medical publications, and the second being the concept of levels of evidence and quality. PMID- 11928802 TI - Randomized clinical trials in breast cancer. AB - Before the second half of this century, treatment approaches to breast cancer were radical and disfiguring. In the past four decades, however, multiple prospective randomized trials have made highly significant advances in the management of patients with this disease. These trials have established, in select patients, breast conservation therapy as a primary therapeutic procedure, and radiation therapy as a means to improve local control and survival. This article provides an overview of some of these trials. PMID- 11928803 TI - Randomized clinical trials in esophageal cancer. AB - The level I evidence leading to current treatment practices in the management of esophageal cancer includes a total of 72 prospective, randomized controlled trials. The impact of these studies are varied. Numerous well-designed surgical trials address important technical issues in resection and reconstruction after esophagectomy; these studies define complication and morbidity rates in addition to overall survival. A large percentage of the prospective, randomized controlled trials address the role of neoadjuvant and adjuvant chemoradiation, with overall survival as the endpoint. The majoritv of these trials are designed to improve on the dismal resectability and overall survival rates in esophageal cancer by initiating novel combination strategies. However, further well-designed PMID- 11928804 TI - Non-enzymatic activities of proteases: from scepticism to reality. PMID- 11928805 TI - Anti-invasive effects of green tea polyphenol epigallocatechin-3-gallate (EGCG), a natural inhibitor of metallo and serine proteases. AB - Several reports have attributed to green tea chemopreventive and therapeutic properties. Epidemiological studies have linked the regular use of green tea to a reduced incidence of breast and colon carcinomas. Tea contains several antioxidants, including polyphenols of the catechin (green tea) and theaflavin (black tea) groups. Green tea derivatives have been shown to act in vitro and in vivo as anti-inflammatory, anti-viral and anti-tumor drugs. Despite the extensive body of data only few studies have investigated the molecular mechanisms underlying these effects. In this brief review we focus on the inhibitory activity of catechins derived from green tea toward proteases involved in tumor invasion. PMID- 11928806 TI - Inhibition of receptor-dependent urokinase signaling by specific Ser to Glu substitutions. AB - We have previously reported that phosphorylation of human urokinase on Ser138/303 abolishes its catalytic-independent motogen and proadhesive abilities, whereas receptor binding is not affected. Here we show that substitution of the two relevant serines with glutamic acid residues impairs the ability of urokinase to mobilize a variety of human and mouse cell lines as well as human primary T lymphocytes. Accordingly, urokinase receptor-dependent signaling, leading to cytoskeletal rearrangements and paxillin re-distribution, does not occur in MCF-7 breast carcinoma cells exposed to 'phosphorylation-like' urokinase. Unlike the wild-type form, di-substituted urokinase is unable to induce the physical association of urokinase receptor with alphavbeta5 vitronectin receptor, which is required for MCF-7 urokinase-dependent cell migration. Finally, the di substituted variant fails to activate p55fgr, a member of the Src tyrosine kinase family, which mediates cell migration and adhesion of U937 myelomonocytic cells. In conclusion, the finding that specific amino acid substitutions strongly interfere with the ability of urokinase to stimulate cell migration, and the associated intracellular events uncover a novel way to regulate urokinase receptor-dependent signaling. PMID- 11928807 TI - Growth factor-dependent proliferation and invasion of muscle satellite cells require the cell-associated fibrinolytic system. AB - The process of muscle regeneration in normal and dystrophic muscle depends on locally produced cytokines and growth factors and requires the activity of the urokinase plasminogen activator/urokinase plasminogen activator receptor/plasminogen activator inhibitor-1 system. In this study we tested the effect of basic fibroblast growth factor (bFGF), hepatocyte growth factor (HGF) and transforming growth factor-beta (TGFbeta) on the fibrinolytic pattern of normal and dystrophic satellite cells, their mitogenic and motogenic activities and the dependence of such activities on the cell-associated fibrinolytic system. We have observed that the urokinase plasminogen activator (u-PA) receptor is weakly upregulated by bFGF in normal satellite cells, while it is strongly up regulated by TGFbeta, mainly in dystrophic myoblasts. bFGF up-regulated u-PA in both normal and dystrophic myoblasts grown in primary culture, while a striking down-regulation was observed with TGFbeta. TGFbeta was the only growth factor able to exceptionally up-regulate plasminogen activator inhibitor-1 (PAI-1), mainly in dystrophic satellite cells. HGF did not show any activity on the fibrinolytic system. Proliferation and invasion into Matrigel matrices of normal and dystrophic cells occurred regardless of the growth factor-dependent regulation of the fibrinolytic system. Nevertheless, each growth factor required the efficiency of the constitutive cell-associated fibrinolytic system to operate, as shown by impairment of growth factor activity with antagonists of u PA and of its receptor. Noteworthy, TGFbeta induced a dose-dependent increase of Matrigel invasion only in dystrophic myoblasts. Since TGFbeta-challenged dystrophic myoblasts undergo an exceptional up-regulation of the receptor and of PAI-1, we propose the possibility that the TGFbeta-induced fibrinolytic pattern (low urokinase plasminogen activator, high receptor and high PAI-1) may be exploited to promote survival and spreading of transplanted engineered myoblasts in Duchenne muscular dystrophy. PMID- 11928809 TI - Random peptide bacteriophage display as a probe for urokinase receptor ligands. AB - The urokinase receptor is a multi-functional protein that plays a central role in cell surface plasminogen activation, cell migration, and cell adhesion. We previously demonstrated that high affinity peptide ligands for the urokinase receptor, which are urokinase competitors, can be obtained from a 15mer peptide library (Goodson et al., 1994). In order to probe for additional urokinase receptor binding sites we affinity selected the same bacteriophage library on complexes of soluble urokinase receptor (suPAR) and the receptor binding domain of urokinase, residues 1-48 (uPA1-48). Bacteriophage were isolated which bound to suPAR and suPAR:uPA1-48 complexes with high yield. The peptide sequences encoded by these bacteriophage were distinct from those obtained previously on urokinase receptor expressing cells, and comprise two groups based upon effects on su-PAR:1 anilino-8-napthalene sulfonate (ANS) fluorescence, and vitronectin binding competition. Alanine scanning mutagensis of the soluble peptides was used to define minimal regions and key residues for suPAR binding by competition with the parent bacteriophage. A comparison of these results with sequences of domains of both vitronectin and integrin alpha-chains, which have been reported to be important for urokinase receptor binding, suggests that the homology with the peptide sequences selected is functionally significant. PMID- 11928808 TI - The col-1 module of human matrix metalloproteinase-2 (MMP-2): structural/functional relatedness between gelatin-binding fibronectin type II modules and lysine-binding kringle domains. AB - Human matrix metalloproteinase-2 (MMP-2) contains three in-tandem fibronectin type II (FII) repeats that bind gelatin. Here, we report the NMR solution structure of the first FII module of MMP-2 (col-1). The latter is described as a characteristic, globular FII fold containing two beta-sheets, a stretch of 3(1) helix, a turn of alpha-helix, and an exposed hydrophobic surface lined with aromatic residues. We show that col-1 binds (Pro-Pro-Gly)6, a mimic of gelatin, with a Ka of approx. 0.42 mm(-1), and that its binding site involves a number of aromatic residues as well as Arg34, as previously found for the second and third homologous repeats. Moreover, the affinity of the in-tandem col-1+2 construct (col-12) toward the longer ligand (Pro-Pro-Gly)12 is twice that for (Pro-Pro Gly)6, as expected from mass action. A detailed structural comparison between FII and kringle domains indicates that four main conformational features are shared: two antiparallel beta-sheets, a central 3(1)-helix, and the quasiperpendicular orientation of the two proximal Cys-Cys bonds. Structure superposition by optimizing overlap of cystine bridge areas results in close juxtaposition of their main beta-sheets and 31-helices, and reveals that the gelatin binding site of FII modules falls at similar locations and exhibits almost identical topological features to those of the lysine binding site of kringle domains. Thus, despite the minor (<15%) consensus sequence relating FII modules to kringles, there is a strong folding and binding site structural homology between the two domains, enforced by key common conformational determinants. PMID- 11928810 TI - Plasmin produces an E-cadherin fragment that stimulates cancer cell invasion. AB - Matrix metalloproteases from the cell surface cleave an 80 kDa E-cadherin fragment (sE-CAD) that induces invasion of cancer cells into collagen type I and inhibits cellular aggregation. Conditioned media from MDCKts.srcCl2 cells at 40 degrees C and 35 degrees C, PCm.src5 and COLO-16 cells at 37 degrees C contained spontaneously released sE-CAD; these 48 h old conditioned media were capable of inhibiting E-cadherin functions in a paracrine way. Here we show direct cleavage of the extracellular domain of E-cadherin by the serine protease plasmin. sE-CAD released by plasmin inhibits E-cadherin functions as evidenced by induction of invasion into collagen type I and inhibition of cellular aggregation. This functional inhibition by sE-CAD was reversed by aprotinin or by immunoadsorption on protein Sepharose 4 fast flow beads with antibodies against the extracellular part of E-cadherin. Our results demonstrate that plasmin produces extracellular E cadherin fragments which regulate E-cadherin function in cells containing an intact E-cadherin/catenin complex. PMID- 11928811 TI - The role of proteases in fibronectin matrix remodeling in thyroid epithelial cell monolayer cultures. AB - Fischer rat thyroid (FRT) cells organize a matrix of extracellular fibronectin (FN) fibrils, which undergoes extensive remodeling according to cell culture confluence. In non-confluent cells FN forms a fibrillar array associated with the ventral cell surface. However, basal FN is progressively removed in confluent cultures and substituted by non-fibrillar FN deposits at lateral cell domains in regions of cell-cell contacts. FRT cells secrete and expose on the plasma membrane the tissue-type plasminogen activator and, in serum-free cultures, plasminogen induces a rapid loss of FN fibrils. Incubation with plasmin inhibitors greatly reduces this effect. FRT cells also express annexin II, a plasminogen receptor, suggesting that plasmin activity is associated with the pericellular enviroment. This is in agreement with the observation that a great reduction in FN degradation is observed if the cells are pre-incubated with carboxypeptidase B, which prevents plasminogen binding to the cells. A gelatinolytic activity with a molecular weigth equivalent to MMP-2 has been demonstrated by zymography of culture media, and the presence of MMP-2 and MT1 MMP on the cell plasma membrane has been detected by immunofluorescence. These results indicate that in the FN remodeling process, occurring during FRT epithelium maturation, both plasmin-dependent (tPA activated) and plasmin independent proteolytic activities are involved. PMID- 11928813 TI - Platelet activating factor inhibits the expression of matrix metalloproteinases and affects invasiveness and differentiation in a system of human neuroblastoma clones. AB - Platelet Activating Factor (PAF), an inflammatory bioactive lipid, has been shown to be involved in the regulation of the activity of matrix metalloproteinases (MMPs). In view of the role played by MMPs in tumor cell invasiveness, we investigated whether PAF influences MMP activity in a system of neuroblastoma clones, the AA5 and AE12 cells, isolated from the human LaN1 neuroblastoma cell line. These clones were characterized by an inverse relationship between invasiveness and differentiative capacity and by the expression of specific cell surface PAF receptors. We found that the levels of mRNAs specific for MMP-2 and for MT1-MMP, the MMP-2 activator, were reduced in both clones treated with 300 nM PAF. These changes are consistent with the reduced secretion and activation of MMP-2 found in the neuroblastoma clones exposed to PAF. These effects were accompanied by an inhibition of invasiveness through Matrigel and by a promotion of differentiation, as revealed by an increased percentage of cells with neurites. The finding that both neuroblastoma clones exposed to the metalloproteinase inhibitors, BB3103 and 1,10-phenanthroline, increased their differentiative capacity and reduced their invasiveness through Matrigel, represents a further indication that PAF modulates differentiation and invasiveness by affecting the activity of MMPs. PMID- 11928814 TI - UVA light stimulates the production of cathepsin G and elastase-like enzymes by dermal fibroblasts: a possible contribution to the remodeling of elastotic areas in sun-damaged skin. AB - Solar elastosis is characterized by accumulation of large amounts of material staining similarly to elastin in the dermis. The nature of this material and the process responsible for its accumulation are still unknown. Elastolytic proteases have important functions in the catabolism of the interstitial matrix and can also generate, by the digestion of the interstitial proteins, soluble peptides which can induce collagen and elastin synthesis and deposition. We investigated whether (i) elastolytic enzymes can be detected in samples from sun-exposed and non-exposed skin, and (ii) ultraviolet (UV) rays influence the production of elastolytic activities in cultured dermal fibroblasts. Immunoelectron microscopy showed a positive reaction for neutrophil elastase and cathepsin G in fibroblast like cells from specimens of sun-exposed areas. Little or no reaction was found in biopsies of sun-protected skin. Fibroblast cultures from sun-exposed skin expressed higher levels of hydrolytic activity against synthetic substrates of elastases and cathepsin G than those obtained from sun-protected areas. Irradiation with UVA strongly stimulated the production of these activities in fibroblasts from sun-protected sites. No significant change was detected in parallel sets of cultures after UVB irradiation. Inhibition experiments indicated that the elastase-like activity expressed by fibroblasts can be attributed to at least two enzymes. PMID- 11928812 TI - Induction of fibronectin mRNA by urokinase- and tissue-type plasminogen activator in human skin fibroblasts: differential role of u-PA and t-PA at the fibronectin protein level. AB - Plasminogen activators of the urokinase- and tissue-type and fetal calf serum (u PA, t-PA, FCS) exert their mitogenic effect on quiescent human dermal fibroblasts and modulate the mRNA expression of cell-cycle related genes. The present study deals with the effects of PAs on the expression of fibronectin (FN), a heterodimeric extracellular matrix (ECM) protein that can be modulated in different ways by various mitogens. The kinetics of FN gene response was examined in quiescent fibroblasts upon PA stimulation (30 min -24 h). The results obtained evidenced that: (i) all mitogens tested (u-PA, t-PA and FCS) led to an increase of FN mRNA expression in early G1, as shown by the analysis of two sequences, III 9, common to all FN mRNAs, and EDA+, present only in the EDA+FN isoform; (ii) the kinetic profiles of FN mRNA stimulation were comparable for the three mitogens, although the effects on the FN-ECM assembly were distinct; (iii) t-PA and FCS led to FN assembly in the ECM, which was absent or decreased in u-PA-treated cultures. Immunobiochemical analysis of total FN and EDA+ FN showed that FN induced by t-PA was mainly dimeric (450-500 kDa), whereas FN induced by u-PA was mainly monomeric (230-250 kDa). These differences are probably due to the differential enzymatic action of t-PA and u-PA on FN, which might be related to a differential role of the two PAs in several physiopathological conditions. PMID- 11928815 TI - The molecular basis for anti-proteolytic and non-proteolytic functions of plasminogen activator inhibitor type-1: roles of the reactive centre loop, the shutter region, the flexible joint region and the small serpin fragment. AB - The serine proteinase inhibitor plasminogen activator inhibitor type-1 (PAI-1) is the primary physiological inhibitor of the tissue-type and the urokinase-type plasminogen activator (tPA and uPA, respectively) and as such an important regulator of proteolytic events taking place in the circulation and in the extracellular matrix. Moreover, a few non-proteolytic functions have been ascribed to PAI-1, mediated by its interaction with vitronectin or the interaction between the uPA-PAI-1 complex bound to the uPA receptor and members of the low density lipoprotein receptor family. PAI-1 belongs to the serpin family, characterised by an unusual conformational flexibility, which governs its molecular interactions. In this review we describe the anti-proteolytic and non proteolytic functions of PAI-1 from both a biological and a biochemical point of view. We will relate the various biological roles of PAI-1 to its biochemistry in general and to the different conformations of PAI-1 in particular. We put emphasis on the intramolecular rearrangements of PAI-1 that are required for its antiproteolytic as well as its non-proteolytic functions. PMID- 11928817 TI - Distinct expression pattern of two related human proteins containing multiple types of protease-inhibitory modules. AB - We have recently identified a gene (the WFIKKN gene) on human chromosome 16 (16p13.3) that encodes a secreted protein containing WAP-type, Follistatin/ Kazal type, Kunitz-type and NTR-type protease-inhibitory modules and an Immunoglobulin domain [Trexler et al., Proc. Natl. Acad. Sci. USA 98 (2001), 3705 - 3709]. In the present work we show that a gene on chromosome 17 encodes a WFIKKN-related protein (WFIKKNRP) that has the same domain organization as the WFIKKN protein. The exon-intron structure of the two genes is also similar as both genes have a single phase 0 intron that splits their WAP domains in equivalent positions. In view of the presence of several protease inhibitory modules in these proteins it seems likely that they serve to control the action of multiple types of proteases. The tissue expression pattern of the two proteins, however, is markedly different suggesting that they have distinct biological roles. Whereas the WFIKKN gene is expressed primarily in adult pancreas, liver and thymus but not in brain and ovary, significant expression of the WFIKKNRP gene is observed in ovary, testis and brain, but not in liver. Pronounced differences could also be seen in the case of fetal tissues: expression of the WFIKKN gene was highest in the lung, skeletal muscle and liver, whereas the WFIKKNRP gene was expressed primarily in brain, skeletal muscle, thymus and kidney. PMID- 11928816 TI - Monocyte-expressed urokinase regulates human vascular smooth muscle cell migration in a coculture model. AB - Interactions of vascular smooth muscle cells (VSMC) with monocytes recruited to the arterial wall at a site of injury, with resultant modulation of VSMC growth and migration, are central to the development of vascular intimal thickening. Urokinase-type plasminogen activator (uPA) expressed by monocytes is a potent chemotactic factor for VSMC and might serve for the acceleration of vascular remodeling. In this report, we demonstrate that coculture of human VSMC with freshly isolated peripheral blood-derived human monocytes results in significant VSMC migration that increases during the coculture period. Accordingly, VSMC adhesion was inhibited with similar kinetics. VSMC proliferation, however, was not affected and remained at the same basal level during the whole period of coculture. The increase of VSMC migration in coculture was equivalent to the uPA induced migration of monocultured VSMC and was blocked by addition into coculture of soluble uPAR (suPAR). Analysis of uPA and uPAR expression in cocultured cells demonstrated that monocytes are a major source of uPA, whose expression increases in coculture five-fold, whereas VSMC display an increased expression of cell surface-associated uPAR. These findings indicate that upregulated uPA production by monocytes following vascular injury acts most likely as an endogenous activator of VSMC migration contributing to the remodeling of vessel walls. PMID- 11928818 TI - Osteopontin modulates prostate carcinoma invasive capacity through RGD-dependent upregulation of plasminogen activators. AB - Osteopontin, a non-collageneous bone matrix protein, is produced in several human tumors but its role in cancer progression has been only partially elucidated. In this study we investigated the potential role of osteopontin in the malignancy of prostate cancer cells. Chemotaxis and chemoinvasion analyses revealed a dose dependent increase in PC3 cell movement induced by osteopontin and a strict dependence of cell invasion on alphavbeta3 integrin function. The pattern of protease expression was modified by osteopontin and was characterized by an upregulation of plasminogen activators. Our findings suggest that osteopontin may confer selective malignant potential to prostate cancer cells through the enhancement of their invasive and proteolytic capability. PMID- 11928819 TI - Expression of matrix metalloproteases (MMP-2, MT1 -MMP) and their tissue inhibitor (TIMP-2) by rat sertoli cells in culture: implications for spermatogenesis. AB - During testicular development and maturation, extracellular matrix (ECM) remodelling is a fundamental process which requires the presence of several proteases and protease inhibitors. Among the proteases, a pivotal role has been proposed for matrix metalloproteases (MMPs) and their tissue inhibitors (TIMPs). Here we report an analysis of MMP-2, MT1-MMP and TIMP-2 expression by rat Sertoli cells in culture using RT-PCR and zymographic techniques. Stimulating Sertoli cells with follicle-stimulating hormone (FSH) in vitro induced evident changes in the level of their mRNA in a time-dependent manner. In the case of TIMP-2 and MT1 MMP, the respective transcripts were augmented up to three-fold after 24 h of treatment, and MMP-2 transcripts increased by four times in the same period. MMP 2 activity determined by gelatin zymography showed an increase in enzyme secretion after FSH stimulation. The results of this study suggest that: (i) at the surface of Sertoli cells pro-MMP-2 activation mediated by MT1-MMP may occur, involving TIMP-2 as a receptor; and (ii) the expression of these molecules is not constitutive in this cell type, but may be modulated by FSH and is therefore implicated in spermatogenesis. PMID- 11928820 TI - Nervous system pathology: the fibrin perspective. AB - Studies of extracellular matrix (ECM) biology in the nervous system have mainly focused on laminin, fibronectin and tenascin-R, proteins that are present during nervous system development and normal function. However, during disease, fibrin, which physiologically is not present in the nervous tissue, is detected at nervous tissue lesions. This review summarizes evidence that correlates fibrin deposition with neuropathology and presents recent findings on cellular mechanisms and intracellular signaling pathways regulated by fibrin that might contribute to nervous system disease. PMID- 11928821 TI - Post-transcriptional regulation of gene expression in the plasminogen activation system. AB - The urokinase-mediated plasminogen activation (PA) system has been shown to play a key role in cell migration and tissue invasion by regulating both cell associated proteolysis and cell-cell and cell-matrix interactions. The expression and activity of the components of this complex system are strictly regulated. The control of the expression occurs both at transcriptional and post-transcriptional levels. This review is focused on the post-transcriptional regulation of gene expression of all components of the PA system. PMID- 11928822 TI - The urokinase plasminogen activator receptor in the regulation of the actin cytoskeleton and cell motility. AB - Cell migration is a complex process requiring tight control of several mechanisms including dynamic reorganization of the actin cytoskeleton and adhesion to the extracellular matrix. The GPI-anchored urokinase plasminogen activator receptor (uPAR) has an important role in the regulation of cell motility in many cell types. This is partly due to the localization of proteolytic activity on the cell surface by binding of the serine protease uPA. Results accumulated over the last decade suggest that uPAR is also involved in motility control through other mechanisms. These include induction of signal transduction events after ligation with uPA, binding to the extracellular matrix molecule vitronectin (VN), and association with integrins and other transmembrane partners. In this review these mechanisms will be discussed with a special emphasis on how the GPI-linked receptor transmits signals to the intracellular milieu and how uPAR participates in the regulation of actin cytoskeleton reorganization and cell adhesion during cell migration. PMID- 11928823 TI - Role of myofibroblasts at the invasion front. AB - Tumor progression occurs within a microecosystem, where cancer cells and myofibroblasts exchange proteinases and cytokines that promote growth directly through stimulation of proliferation and survival, as well as invasion through local proteolysis of the extracellular matrix and stimulation of motility. Myofibroblasts maintain the capacity of fibroblasts to induce differentiation. Fibroblasts are the main source of tumor-associated myofibroblasts. The transition to myofibroblasts also occurs in noncancerous situations. This transition is modulated by mechanical stress and cytokines, amongst which transforming growth factor-beta. The cross-talk between cancer cells and myofibroblasts illustrates the microecosystem of tumor invasion. In order to consider myofibroblasts as a possibly new target for cancer therapy, further characterization of the molecular cross-talk between myofibroblasts and cancer cells is required. PMID- 11928824 TI - Transmodulation of cell surface regulatory molecules via ectodomain shedding. AB - Cell responses to exogenous stimuli often result in a rapid decrease of cell surface density of a wide range of diverse regulatory proteins, receptor and adhesion molecules in particular. This decrease may occur in a ligand-dependent fashion (down-regulation), following endocytosis and degradation by lysosomal proteases, or by down-modulation, where molecules are targeted by endoproteases directly on cell surface. These proteases are recruited by trans-modulating agents, different from ligand, which act via their own receptors and the related intracellularly-generated signals. Endoproteolytic activity determines the release of large portions (shedding) of substrate proteins, called ectodomains, which are usually not ligand-bound, and therefore represent biologically-active molecules. Ectodomain shedding is involved in a number of pathophysiological processes, such as inflammation, cell degeneration and apoptosis, and oncogenesis. Common features of the process, such as the involvement of protein kinase C and of transmembrane metalloproteases, have been identified. In this review, we summarize basic concepts on down-modulation and ectodomain shedding, and provide an update of the issue with respect to: (i) new entries to the list of molecules found involved in the process; (ii) current views about the upstream control of shedding, i.e. the pathways linking the signals triggered by the trans modulating agents to the activation of endoproteolytic activity on the cell surface. PMID- 11928825 TI - The plasminogen activating system in periodontal health and disease. AB - The plasminogen activating system is important for extracellular proteolysis and plays a regulatory role in interactions with other tissue degrading systems. Studies on the plasminogen activating system in gingival crevicular fluid (GCF) as well as gingival tissue are reviewed. t-PA, u-PA, PAI-1 and PAI-2 have all been detected in GCF. Especially t-PA and PAI-2 are found in high concentrations. In tissue studies fibrinolytic activity has been found in the gingival pocket epithelium in humans and in animal studies. t-PA and PAI-2 have been detected there immunohistochemically. Local production of the PAs and PAls has been verified with in situ hybridization. In inflammation, a more intense and widespread immunohistochemical staining of t-PA and PAI-2 is seen. Higher concentrations of t-PA and PAI-2 are found in GCF but the balance between them seems to be constant. A systemically disturbed balance of the plasminogen activating system in GCF has been observed during pregnancy, with a possible protective function of PAI-2. In studies of periodontitis, the production of PAI 2 seemed to be locally lowered at impaired sites. In a study of children, a higher inflammatory response to bacterial plaque was accompanied by a higher fibrinolytic ativity in GCF samples. Bacterial LPS has been found to change the ratio of t-PA to PAI-2 in cultured gingival fibroblasts. Interactions between PAI 2 and a protease in the gingival epithelium has been verified through the immunohistochemical detection of relaxed PAI-2. PMID- 11928826 TI - Apolipoprotein(a): structure-function relationship at the lysine-binding site and plasminogen activator cleavage site. AB - Apolipoprotein(a) [apo(a)] is the distinctive glycoprotein of lipoprotein Lp(a), which is disulfide linked to the apo B100 of a low density lipoprotein particle. Apo(a) possesses a high degree of sequence homology with plasminogen, the precursor of plasmin, a fibrinolytic and pericellular proteolytic enzyme. Apo(a) exists in several isoforms defined by a variable number of copies of plasminogen like kringle 4 and single copies of kringle 5, and the protease region including the backbone positions for the catalytic triad (Ser, His, Asp). A lysine-binding site that is similar to that of plasminogen kringle 4 is present in apo(a) kringle IV type 10. These kringle motifs share some amino acid residues (Asp55, Asp57, Phe64, Tyr62, Trp72, Arg71) that are key components of their lysine binding site. The spatial conformation and the function of this site in plasminogen kringle 4 and in apo(a) kringle IV-10 seem to be identical as indicated by (i) the ability of apo(a) to compete with plasminogen for binding to fibrin, and (ii) the neutralisation of the lysine-binding function of these kringles by a monoclonal antibody that recognises key components of the lysine binding site. In contrast, the lysine-binding site of plasminogen kringle 1 contains a Tyr residue at positions 64 and 72 and is not recognised by this antibody. Plasminogen bound to fibrin is specifically recognised and cleaved by the tissue-type plasminogen activator at Arg561-Val562, and is thereby transformed into plasmin. A Ser-Ile substitution at the activation cleavage site is present in apo(a). Reinstallation of the Arg-Val peptide bond does not ensure cleavage of apo(a) by plasminogen activators. These data suggest that the stringent specificity of tissue-type plasminogen activator for plasminogen requires molecular interactions with structures located remotely from the activation disulfide loop. These structures ensure second site interactions that are most probably absent in apo(a). PMID- 11928827 TI - The revised Helsinki Declaration: good for patients? Good for public health? PMID- 11928828 TI - Incidence of cardiovascular disease, cancer and death in postmenopausal women affirming use of hormone replacement therapy. AB - AIM: The goal of this study was to evaluate the incidence of myocardial infarction, cancer and death in relation to use of hormone replacement therapy (HRT). METHODS: Nine years' follow up of an urban cohort of peri-/postmenopausal women was undertaken. Local and national registers were used for retrieval of events. RESULTS: The incidence of myocardial infarction per 1.000 person-years in users and non-users was 0.61 (5/962) and 2.20 (92/4759) respectively, adjusted relative risk (RR) 0.37; 95% confidence interval 0.15-0.90. Rates of mortality from cardiovascular disease and cancer were 0.36 and 1.10, p= 0.058, and 2.60 and 2.09, p=0.360 respectively. In terms of all-cause mortality the adjusted RR was 1.02; 0.69-1.52, incidence of cancer 1.28; 1.01-1.64, breast cancer 1.52; 1.01 2.28 and endometrial cancer 3.61; 1.54-8.46. CONCLUSIONS: Women affirming use of HRT had a lower incidence of myocardial infarction. Further studies are needed to assess whether the absence of effect on total mortality may be accounted for by an increased cancer risk. PMID- 11928829 TI - Prevalence of subjective health complaints (SHC) in Norway. AB - AIMS: As subjective health complaints are one of the major reasons for short- and long-term sickness absence it is important to know the prevalence of these conditions in the general population. METHODS: In this cross-sectional study, 1,240 individuals (aged 15-84 years) from the normal population in Norway answered the Subjective Health Complaint (SHC) inventory in spring 1996. RESULTS: The prevalence of reporting subjective health complaints was high: 80% reported musculoskeletal complaints, 65% reported "pseudoneurological" complaints (tiredness, depression, dizziness), 60% gastrointestinal complaints, 34% allergic complaints, and 54% flu-like complaints. The prevalence of substantial complaints was low: 13% reported musculoskeletal complaints, 5% reported "pseudoneurological" complaints, 4% gastrointestinal complaints, 2% allergic complaints, and 18% flu-like complaints. Women had higher prevalence of musculoskeletal, "pseudoneurological", and allergic complaints compared with men, and reported more substantial complaints on all subscales. Individuals older than 50 years were less likely to report headache, tiredness, eczema, and flu-like complaints compared with individuals younger than 30 years. However, they had higher risk of arm pain, shoulder pain, palpitations, and several gastrointestinal and allergic complaints. The intensity of musculoskeletal, gastrointestinal, and allergic complaints was significantly higher in the oldest age group. CONCLUSIONS: Subjective health complaints are very common in the normal population; there are gender and age differences in both prevalence and degree of complaints. The intensity of subjective health complaints forms a continuum, and there are no obvious cut-off point to indicate what are "normal" complaints and what is illness. PMID- 11928830 TI - Locus of control in relation to smoking cessation during pregnancy. AB - AIMS: This study was carried out to investigate how the locus of control (LOC) varies between those who stop smoking in connection with pregnancy and those who do not. The LOC reflects the extent to which a person is in control of her or his life events. METHODS: One hundred and two women who were regular smokers at the time of conception and paid their first visits to the antenatal clinic (ANC) on account of the current pregnancy participated. The study was quantitative and questionnaire based, covering background factors, smoking habits, and the LOC. One hundred of the women were met a second time after the delivery. Smoking habits were monitored by the content of carbon monoxide in expired air. The LOC was recorded on a VAS scale (0-10). A lower index was consistent with a more internal LOC and a higher with a more external LOC. RESULTS: The median LOC found was 4.0 (range 1.1-7.9). The LOC was not correlated to the CO in expired air. Younger age was linked to a more internal LOC, while worse sleep was correlated to a more external LOC (p<0.05). No differences were found between those who stopped smoking and those who did not. This was the case in both light and heavy smokers. CONCLUSION: The LOC does not seem to influence the ability to stop smoking during pregnancy. PMID- 11928831 TI - Changes in community and individual level psychosocial coronary risk factors in the Danish MONICA population, 1982-92. AB - AIM: Myocardial infarction incidence declined by 3-5% per year from 1982-92 in the Danish study population of the WHO MONICA project. This study examined whether psychosocial coronary risk factors changed in the population during this period. METHODS: Cross-sectional data from 6,695 Danish men and women aged 30, 40, 50, and 60 years, were examined in 1982-84, 1986-87, and 1991-92. Data from Social Registers were used to analyse trends in community social structure. RESULTS: From 1982 to 1992 an increase in individuals' school education and social class followed an increase in the general welfare of the study area. At the same time type A behaviours increased in both men and women, while there were no changes in reported personal problems. The proportion of women living alone increased, while the proportion of emotionally distressed decreased. In men there were no changes in cohabitation status or emotional distress. The changes in the psychosocial factors were the same in different educational groups. CONCLUSION: The socioeconomic position of the population improved at the individual and the community level during the study period. This was concurrent with the declining incidence of myocardial infarction. PMID- 11928832 TI - Limiting long-term illness and the experience of financial strain in Sweden. AB - AIMS: This study aims to investigate whether those individuals exposed to certain socioeconomic events (e.g. unemployment or early retirement) are more at risk of experiencing financial strain if they are also exposed to some limiting long-term illness. METHODS: This is a cross-sectional study using a Swedish sample of 41,497 individuals between the ages of 25 and 65 selected from the Survey of Living Conditions of Statistics Sweden. The measure of association used in the study is the prevalence odds ratio, and interaction analyses are performed in order to assess differential susceptibility to experience financial strain. RESULTS: The results show that young persons, living in poor households, unemployed or early retired, reporting a high degree of impairment, present a greater probability of experiencing financial strain. Severe impairment has proved to be a component in the possible causal chain leading to the experience of financial strain, which interacts with the factors studied, increasing significantly the probability of certain groups experiencing financial hardship. CONCLUSION: The results sustain the hypothesis that social consequences of illness, such as financial strain, might contribute to aggravate inequalities between socioeconomic groups under certain conditions such as unemployment and poverty. PMID- 11928833 TI - Lifestyle factors and choice of hormone replacement therapy among Danish nurses. AB - BACKGROUND: Studies have found that women who use hormone replacement therapy (HRT) are healthier or report better health than non-users. This study examines the association between lifestyle factors, self-reported health and the use of HRT in Danish female nurses above the age of 50 years. METHODS: Postal questionnaires were used for data collection. Two cohorts were examined, a younger including women between 50 and 59 years and an older including women of 60 years and above. RESULTS: Of the 14,071 nurses included, 5,528 (39%) reported ever using HRT. In both cohorts, ever using HRT was positively associated (odds ratio > 1) with intake of vitamin supplements, alcohol consumption at weekends, consulting a physician, use of medicine, and previous use of oral contraceptives. Only a few differences were found between former and current users of HRT. A highly significant association was found between ever using HRT and self-reported poor health. CONCLUSIONS: In this study use of HRT was not found to be associated with a particularly healthy lifestyle or better health. Therefore, these factors should be specifically considered in studies evaluating the effect of HRT. PMID- 11928834 TI - Predicting the effect of prevention of ischaemic heart disease. AB - BACKGROUND: Priority setting in public health policy must be based on information on the effectiveness of alternative preventive and therapeutic interventions. The purpose of this study is to predict the effect on mortality from ischaemic heart disease (IHD) in Denmark of reduced exposure to the risk factors hypertension, hypercholesterolaemia, cigarette smoking, and physical inactivity. METHODS: The simulation model, "Prevent", was used. The input data for the model were population size, death rates, prevalence of cardiovascular risk factors, and relative risks. Scenarios of the impact of reduced exposure to risk factors on mortality from IHD were studied, including the health benefits of successful implementation of some of the targets in the Danish Government programme for public health and health promotion, 1999-2008. RESULTS: The greatest reduction in mortality from IHD would be achieved by interventions against cigarette smoking. If the proportion of cigarette smokers could be reduced by one-third within 10 years, the number of deaths due to IHD among people under 65 would be 10% lower for men and 15% lower for women. If the proportion of heavy smokers or the proportion of people with hypertension were reduced by 25%, the rate of mortality from IHD would be 5% lower for men and 6-7% lower for women. Reducing the number of cases of severe hypercholesterolaemia by 25% would lower the IHD mortality rate before age 65 by 3% for men and 6% for women after 15 years, similar to the effect of easy exercise for at least 4 hours a week among physically inactive people. PMID- 11928835 TI - Falls among frail older people in residential care. AB - AIMS: A prospective study was carried out to investigate the incidence, circumstances, and injuries from falls among frail older people living in three different types of Swedish residential care settings. METHODS: The settings were senior citizens' apartments, an old people's home, and a group dwelling for people with dementia. The falls were registered during the three-year study period on a semi-structured fall report, and injurious falls were categorized according to severity. RESULTS: In total 428 falls occurred among 121 residents. The incidence rate of falls at the group dwelling was twice the rates of the old people's home and senior citizens' apartments (4282 compared with 1709 and 2114 falls per 1000 person-years respectively). Some 27% of the falls occurred during the night (2100h to 0600h) and 28% were related to a visit to the lavatory. The presence of acute disease at the time of a fall was diagnosed in 23% of the falls. Some type of injury occurred in 118 falls (28%) and 36 of these (8%) led to moderate or serious injuries. In total 48 fractures were diagnosed. CONCLUSIONS: In a preventive programme for falls and injuries in residential care settings, areas of particular interest should include falls after mealtimes and falls at night, conditions of acute diseases, rising up from sitting, walking, and activities in progress, especially visits to the lavatory. PMID- 11928836 TI - Boys and girls smoking within the Danish elementary school classes: a group-level analysis. AB - AIMS: To quantify the correlation between male and female smoking prevalence in elementary school classes by group-level analysis. METHODS: This study was the Danish contribution to the cross-national study Health Behaviour in School-Aged Children (HBSC) 1998. Ninety school classes at grade nine (1,515 students) from a random sample of schools in Denmark took part. The proportion of male and female "at all" smokers and daily smokers in the school class was calculated. RESULTS: The mean "at all" smoking proportion in the school classes is 39% for girls and 32% for boys. The proportion of male and female smokers within school classes does not correlate. There is high variation in male and female smoking behaviour between school classes. CONCLUSIONS: The influence of social classroom environment on the processes causing smoking behaviour may be different for boys and girls. This paper illustrates that group-level analysis provides valuable new knowledge. PMID- 11928837 TI - Changes in health beliefs after labelling with hypercholesterolaemia. AB - BACKGROUND: Patients' health beliefs influence their willingness to comply with medical advice. In an earlier study, it was found that men with a previous history of information on risk factors for ischaemic heart disease expressed more feelings of threat to their health than did men without this experience. As anxiety may have adverse effects, such as making patients avoid the desired action, this could complicate adequate patient treatment. AIMS: To investigate the impact on health beliefs caused by participation in a screening programme for risk factors for ischaemic heart disease, including individualized information to patients with hypercholesterolaemia. METHODS: A random sample of middle-aged, urban men participating in a health screening completed a questionnaire on socioeconomic factors, medical history, lifestyle, and health beliefs. Blood pressures and plasma cholesterol values were measured. Four months after the initial screening, hypercholesterolaemic men and controls completed the questionnaire again. RESULTS: In a univariate analysis, no differences in health belief indices were found between cases and controls at the baseline screening. Controls achieved lower values of the indices "perceived control over illness" and "medical motivation" at follow-up. In a matched case-control design, the differences in "medical motivation" increased between cases and controls because controls reported lower values. "Perceived threat to health" did not change, and it is suggested that this is due to the supportive information to the patients. CONCLUSION: Individualized and supportive patient information on risk factors for cardiovascular disease does not increase patients' perceptions of threat. PMID- 11928838 TI - Clinical manifestations and laboratory assessment in an enterovirus 71 outbreak in southern Taiwan. AB - An epidemic of enterovirus 71 (EV71) infection compatible with hand, foot and mouth disease and associated with high morbidity and mortality occurred in Taiwan in 1998. We recruited 90 patients (50 males, 40 females) with definite EV71 infections for clinical and laboratory analysis. The neurological signs and symptoms, all of which occurred during the febrile period, in patients with central nervous system (CNS) involvement (aseptic meningitis, encephalitis or myelitis) were myoclonic jerks (23/33), vomiting (10/33), ataxia (7/33), lethargy (6/33), seizure (4/33) and tremor (2/33). Patients with CNS involvement had longer durations of fever (4.6+/-0.2 vs. 3.1+/-0.3 d; p <0.01) and a higher white blood cell count (12,512+/-658 vs. 10,607+/-409 cells/mm3; p = 0.01) than patients without CNS involvement. The case fatality rate in patients with CNS involvement was 4/33 (12%), whereas no fatalities (0/57) occurred in patients without CNS involvement. Six of 11 patients subjected to MRI showed a high intensity T2-weighted signal in the brainstem. A nested fluorescent RT-PCR for detection of virus in throat and stool specimens showed higher sensitivity than viral culture. Viremia was detectable using RT-PCR in 20% of cases (3/15), whereas no virus was isolated from culture or detected by RT-PCR in cerebrospinal fluid. PMID- 11928839 TI - Excellent booster response 4-6 y after a single primary dose of an inactivated hepatitis A vaccine. AB - We studied the immune response of an inactivated hepatitis A vaccine (Havrix 1440) given to middle-aged travellers 4-6 y after a single, primary dose. Anti HAV antibodies in serum were checked before and 28-35 d after the booster. All 25 vaccinees showed an impressive anamnestic booster response (geometric mean titres 32 and 2993 mIU/ml before and after the booster, respectively). The study confirms experimental data indicating that I dose of inactivated hepatitis A vaccine induces a long-term proliferative T-cell response in addition to producing anti-HAV antibody. As recall memory for this vaccine is elicited several years after a single dose there is probably no need for a second vaccine dose. PMID- 11928840 TI - Polymorphism of the IL-10 gene is associated with susceptibility to herpes zoster. AB - Herpes zoster (HZ) is induced by diseases or medical treatments leading to a decrease in cell-mediated immunity, but in most cases it appears without any detectable, predisposing condition. We report here that susceptibility to HZ is genetically determined. The promoter region of the IL-10 gene contains 3 single nucleotide polymorphisms (G/A at -1082, C/T at -819 and C/A at -592) which form 3 haplotypes, GCC, ACC and ATA. These haplotypes are known to differ in terms of their effect on the transcription of the IL-10 gene. In a cohort of 60 HZV patients, the number of carriers of the ATA haplotype was 32 (53%), while in blood donors the frequency was 151/400 (38%) (p < 0.05). PMID- 11928841 TI - Epidemiological investigation of a food-borne gastroenteritis outbreak caused by Norwalk-like virus in 30 day-care centres. AB - In March 1999, an outbreak of gastroenteritis occurred affecting 30 day-care centres served by the same caterer. A retrospective cohort study was performed in 13 randomly selected day-care centres to determine the source and mode of transmission. Electron microscopy and PCR were used to verify the diagnosis. The overall attack rate (AR) was 37% (195/524): 30% in children and 62% in adults. Modified by the age of the patient, eating pumpkin salad served on 1 March was associated with becoming an early case (odds ratio = 3.9; 95% confidence interval 1.8-8.8). No significant association was found between food consumption and becoming a late case. The primary food-borne AR was 27% and the secondary AR was 14%. The same genotype of Norwalk-like virus was found in 5 cases and in 1 ill and 1 asymptomatic food-handler. Contamination by 1 of the food-handlers seems the most likely route of spread of the virus and underlines the importance of strict hygienic routines. PMID- 11928842 TI - Synercid plus vancomycin for the treatment of severe methicillin-resistant Staphylococcus aureus and coagulase-negative staphylococci infections: evaluation of 5 cases. AB - Synercid (quinupristin/dalfopristin), the first semi-synthetic injectable streptogramin, is a promising alternative to glycopeptides against many Gram positive multiresistant bacteria. Vancomycin is still considered an effective agent for the treatment of methicillin-resistant Staphylococcus aureus (MRSA) infections but therapeutic failures with glycopeptides have been observed, even for the treatment of infections caused by S. aureus strains sensitive to vancomycin. Synercid, in combination with a glycopeptide, may address this problem without causing significant side effects due to the different toxicity patterns of the 2 antimicrobials. This study reports our experience with the combination of Synercid and vancomycin in 5 patients with severe infection caused by MRSA or methicillin-resistant coagulase-negative Staphylococcus. PMID- 11928843 TI - High serum hepatocyte growth factor levels in the acute stage of community acquired infectious diseases. AB - Acute serum levels of hepatocyte growth factor (HGF) were studied in 6 clinical groups with (i) gastroenteritis, (ii) skin and soft tissue infection, (iii) urinary tract infection, (iv) septicemia, (v) influenza, and (vi) chronic hepatitis C in comparison with a normal control group using an enzyme-linked immunosorbent assay method. We found that serum HGF levels were significantly higher in patients with acute infectious diseases (p < 0.0001) compared to patients with chronic viral hepatitis and healthy controls. Serum HGF and CRP levels were correlated significantly (r=0.65, p < 10(-7)). We conclude that serum HGF levels are elevated in patients with acute infectious diseases. PMID- 11928844 TI - Infective endocarditis after transrectal prostatic biopsy. AB - We report the first case of infective endocarditis secondary to transrectal prostatic biopsy, occurring 2 weeks later in a patient with no obvious risk factors. Enterococcus faecalis was isolated in blood and urine cultures. This case suggests that infective endocarditis can be associated with transrectal procedures. PMID- 11928845 TI - Yersinia enterocolitica O:3 infection of a prosthetic knee joint related to recurrent hemarthrosis. AB - Yersinia enterocolitica was isolated in joint fluid containing blood extracted from the knee of an 80-y-old woman with a 10-y history of total knee arthroplasty. Recurrent hemarthrosis had previously occurred in this knee. It appeared that the effusion of blood led to the deposition of iron on the joint, which may have contributed to the development of infection. PMID- 11928846 TI - Medically treated splenic abscess due to Brucella melitensis. AB - Brucellosis may lead to complications that affect different organ systems, including the liver and spleen. In acute disease, hepatosplenic abscess is a rare complication. We report herein a woman with splenic abscess due to acute Brucella melitensis infection who was successfully treated with antibiotics alone. PMID- 11928847 TI - Lactobacillus rhamnosus septicaemia in a patient with a graft in the inferior vena cava. AB - A case of septicaemia that repeatedly showed growth of Lactobacillus rhamnosus in blood cultures is reported. The patient improved after removal of a Goretex patch in the inferior vena cava that was the focus of the infection. Lactobacilli can, in certain settings, be the cause of clinically important infections. PMID- 11928848 TI - Sinusitis due to Stenotrophomonas maltophilia. AB - This is the first report of a patient diagnosed with sinusitis due to Stenotrophomonas maltophilia. Despite the organism being resistant to trimethoprim-sulfamethoxazole, the infection was cured by drainage and treatment with intravenous piperacillin, followed by oral ciprofloxacin. PMID- 11928849 TI - Pasteurella multocida septicaemia in 2 Swedish patients. AB - The clinical manifestation of human infections with Pasteurella multocida is most often cellulitis and this pathogen rarely causes septicaemia. We describe 2 Swedish patients with P. multocida septicaemia who were admitted to the same ward within the space of 7 months. PMID- 11928851 TI - Coccidioidal meningitis: incidental diagnosis 3 y after ventriculo-peritoneal shunt placement for hydrocephalus. AB - A patient from the American Midwest developed hydrocephalus after a trip to southwest California, necessitating placement of a ventriculo-peritoneal shunt. Coccidioidal meningitis was diagnosed incidentally 3 y later, during evaluation for shunt malfunction. PMID- 11928850 TI - Pichia ohmeri prosthetic valve endocarditis and review of the literature. AB - Fungal prosthetic valve endocarditis (PVE) is a serious complication of valve replacement surgery. We report the first case of documented Pichia ohmeri PVE in an immunocompetent man who was successfully treated with valve replacement and antifungal therapy with amphotericin B. PMID- 11928852 TI - Breast implant infection caused by Trichosporon beigelii. AB - We describe herein an immunocompetent patient who developed a breast implant infection caused by Trichosporon beigelii. To our knowledge, this is the first reported case of such an infection. PMID- 11928854 TI - Acute Aspergillosis gastritis in a case of fatal aplastic anemia. AB - A rare case of stomach perforation following acute aspergillosis gastritis in the course of a fatal severe aplastic anemia is reported. PMID- 11928853 TI - Candida albicans endocarditis diagnosed by PCR-based molecular assay in a critically ill pediatric patient. AB - A young Down's syndrome patient developed tricuspid valve endocarditis several years after undergoing surgical closure of a congenital ventricular septal defect. Fungal etiology was established by PCR amplification of the Candida albicans ERG11 gene. Although antifungal therapy was administered, surgical replacement of the infected valve was required to eliminate the infection. PMID- 11928855 TI - A case of zygomycosis and invasive candidiasis involving the epiglottis and tongue in an immunocompromised patient. AB - Invasive fungal infections are associated with high morbidity and mortality in immunocompromised patients. We describe an unusual case of concomitant invasive candidiasis and zygomycosis of the tongue and epiglottis that occurred in a young patient with neutropenia during chemotherapy for acute myelogenous leukemia and was successfully treated medically. PMID- 11928856 TI - Visceral leishmaniasis with pericarditis in an HIV-infected patient. AB - The clinical presentation of visceral leishmaniasis, or kala-azar, is variable but usually includes fever, severe cachexia, lymphadenopathy and hepatosplenomegaly. In immunocompromised patients the clinical course of the disease is even less specific and the diagnosis is often made by means of incidental detection of the parasites at atypical sites such as the gastrointestinal tract, peripheral blood, lungs and cerebrospinal fluid. We describe a case of pericardial leishmaniasis in an HIV-infected patient. PMID- 11928857 TI - Three cases of Rothia dentocariosa bacteraemia: frequency in Denmark and a review. AB - Three cases of non-endocarditis-associated Rothia dentocariosa bacteraermia occurred in Viborg County within the space of a year, whereas no cases have been registered in other parts of Denmark during the last 10 y. Two patients wore dentures but had no history of oral infection, while in the last patient a tooth abscess was detected on examination. R. dentocariosa is a common inhabitant of the oral cavity and the causative agent of a wide spectrum of infectious symptoms. It is only rarely identified in blood cultures, possibly due the inadequacy of the available identification methods. PMID- 11928858 TI - Group G streptococcal bacteraemia: an opportunistic infection associated with immune senescence. AB - The number of cases of group G streptococcal bacteraemia reported worldwide is increasing. Twenty-six cases of group G streptococcal bacteraemia were identified during a 70-month period at a single university teaching hospital in Sheffield, UK. These cases represented 20% of all bacteraemias due to beta-hemolytic Streptococci, a higher proportion than previously reported. The median age of these cases was 72 y and although medical comorbidities were common only cutaneous ulceration was clearly linked to the presenting syndromes. The skin was the source of infection in 16 cases (62%) and the most frequent clinical presentations were cellulitis in 13 cases (50%) and endovascular infection in 5 (19%). Eight (31%) of the cases died during the period of follow-up but only 2 deaths were related to the streptococcal infection. Immunosenescence represents the major risk factor for group G streptococcal infection in this population and comorbidities, including carcinoma, may be markers of the senescent immune system rather than direct contributing factors to group G streptococcal bacteraemia. PMID- 11928859 TI - Nosocomial transmission of syphilis during haemodialysis in a developing country. AB - Patients with end-stage renal disease (n = 187) secondary to diverse aetiologies who underwent haemodialysis (HD) between November 1996 and November 2000 were routinely screened for syphilis using the rapid plasma reagin (RPR) test and confirmed by means of a microhemagglutination assay for Treponema pallidum. All the confirmed syphilis patients were asymptomatic and were diagnosed serologically. A true seroprevalence of 6.9% (13/187) and a biological false seropositivity of 4.2% (8/187) for syphilis were recorded. Most (11/13) of the true seropositive patients were aged between 51 and 80 y. Whilst 10/13 patients were true syphilis seropositive at the time of first HD, 3/13 patients became true seropositive an average of 12 months (range 10-14 months) after HD. Penicillin treatment was given to all 13 patients simultaneously only after the appearance of 3 new true syphilis seropositive cases. Complete seroreversion was observed in 4/10 patients in the pre-HD true syphilis seropositive group of presumptive transmitters who became RPR-negative, whereas the 3 new true seropositive cases showed a serial 4-fold decline in RPR titres 12 months after penicillin therapy, suggestive of an active disease with adequate therapeutic response. These results clearly indicate that latent syphilis is prevalent in long-term elderly HD patients. The true seroconversion of 3 new patients who had undergone HD for an average of 12 months is indicative of nosocomial transmission and the silently active nature of the disease, which necessitates regular monitoring of syphilis serology among HD patients. PMID- 11928860 TI - Seroepidemiology of Rickettsia africae infection in Norwegian travellers to rural Africa. AB - Rickettsia africae is the causative agent of African tick bite fever (ATBF), an acute febrile illness frequently accompanied by inoculation eschars, regional lymphadenitis, myalgia and severe headache. Recently, ATBF has been recognized as an emerging health problem for international travellers to rural sub-Saharan Africa. To estimate the incidence, risk factors for and proportion of symptomatic cases of travel-associated R. africae infection, we performed a seroepidemiological study of 152 first-time Norwegian travellers to rural areas in sub-Equatorial Africa. Seropositivity was based on the detection of specific antibodies to R. africae in microimmunofluorescence and/or Western blotting assays. Thirteen (8.6%) travellers were seropositive to R. africae. Eight (62%) seropositive travellers reported symptoms consistent with ATBF; of these, 2 had received antirickettsial therapy. Using multiple logistic regression, the following factors were found to be significantly associated with seropositivity: hunting as the purpose of travel [odds ratio (OR) 10.1; 95% confidence interval (CI) 1.5-69; p=0.019] and stay in rural areas of > 7 d (OR 6.0; 95% CI 1.5-24; p=0.012). This first seroepidemiological study on travel-associated R. africae infection suggests that the infection may be common in international travellers to rural sub-Saharan Africa but that most cases are asymptomatic or clinically mild and self-limited. PMID- 11928862 TI - Giant cystic pheochromocytoma located in the renal hilus. AB - A malignant tumor in the past medical history of a patient often makes the differential diagnosis of a second tumor more difficult, especially if one of the tumors does not show its characteristic features. The authors report a case of a 55-year-old male who presented with a malignant melanoma on his left shoulder. A retroperitoneal giant cystic mass, 200 mm in diameter, was found incidentally. Adrenal origin was ruled out by imaging techniques. The absence of typical clinical symptoms made a correct preoperative diagnosis unlikely, and severe cardiovascular complications set in during surgery. Considering the characteristics of the cutaneous malignant melanoma, the metastatic origin of the giant retroperitoneal tumor was not likely either. During surgery the left kidney, with a cystic tumor located in the hilus, was removed. The postoperative pathologic diagnosis was pheochromocytoma located in the hilus of the left kidney. PMID- 11928861 TI - Circulating antibodies to lipoarabinomannan in relation to sputum microscopy, clinical features and urinary anti-lipoarabinomannan detection in pulmonary tuberculosis. AB - An enzyme-linked immunosorbent assay (ELISA)-based investigation of anti lipoarabinomannan (LAM) antibody levels in the sera of patients with acid-fast bacilli (AFB)-positive pulmonary tuberculosis (PTB), AFB-negative PTB and non-TB respiratory tract symptoms was conducted. The anti-LAM results were further evaluated using urine LAM detection and a clinical diagnostic score (DS) system as references. Using sputum AFB as a reference, positive anti-LAM was found in 66.9% of 139 AFB-positive PTB, 34.4% of 61 AFB-negative PTB and 23.5% of 800 non TB patients and in 8% of 50 healthy individuals. The positive and negative predictive values were 48.7% and 87.4%, respectively. Using the DS as a reference, the sensitivity and specificity were 50.5% and 78.3%, respectively, whereas 45.8% of urine LAM positives and 77.9% of urine LAM negatives were correctly identified by the anti-LAM ELISA. In TB endemic areas a negative anti LAM could be of practical value, particularly when other indicators of PTB are negative. Using any of these methods as a reference, a positive anti-LAM would mislead in about one-quarter of cases. Had all the 3 methods been combined and at least 2 positive tests sufficed, 90.6% of AFB-positive PTB, 52.5% of AFB-negative PTB and 94.9% of non-TB patients would have been correctly diagnosed. Apart from the possible impact of HIV, the low accuracy of the current assay could be due to intravascular formation of LAM-anti-LAM complexes, latent TB or environmental mycobacterial infections. PMID- 11928863 TI - Oncocytic meningioma: a case report. AB - Oncocytic meningioma is a recently described rare variant of meningothelial neoplasms that typically occurs as a large cell tumor with granular cytoplasm. The distinct histological features of numerous cells with granular cytoplasm and the ultrastructural evidence of numerous mitochondria in the cytoplasm differentiate this tumor from other neoplasms with granular appearance. We report an additional case of oncocytic meningioma investigated by ultrastructural and immunohistochemical methods. PMID- 11928864 TI - Endocervicosis of the urinary bladder. Immunohistochemical comparative study between a new case and normal uterine endocervices. AB - Endocervicosis of the urinary bladder is a rare non-neoplastic condition characterized by endocervical-type glands deeply situated in the urinary bladder wall of women of reproductive age. We compared the immunohistochemical phenotype of a case of endocervicosis in a 35-year-old woman with four normal uterine endocervices. We tested antibodies known as reactive in the uterus and not mentioned or negative in the urothelium (HBME-1, estrogen receptor (ER), progesterone receptors (PR), DF3, Chromogranin). The proliferative index was assessed with MIB-1 antibody. Endocervicosis glands displayed stronger expression of HBME-1, ER and PR than normal endocervices, while the urothelium was negative. There was no difference in DF3 expression. The number of Chromogranin-positive cells was higher in endocervicosis than in the endocervices. The proliferative index was higher in the endocervicosis glands (15%) than in the normal endocervices (mean 3%), but was within the normal range established for endocervical glands. Our results confirm the endocervical nature of endocervicosis and constitute further arguments for the mullerian origin hypothesis. The only modestly increased proliferative index, as compared to endocervical malignancies, is consistent with a benign diagnosis. PMID- 11928865 TI - Clear cell carcinoma arising in endometriosis of the rectum following progestin therapy. AB - A 40-year-old woman presented at our hospital with rectal stenosis. Computed tomography demonstrated a conglomerate tumor in the pelvis and malignant look alike lesions in the liver and in both lungs. A palliative deep rectum resection was done. Histologically, clear cell carcinoma with a small area of endometrioid carcinoma was diagnosed. Severe endometriosis had been diagnosed 13 years earlier. The patient had been treated with medroxyprogesterone acetate (MPA) for two years; no estrogen therapy had been given. The association between unopposed estrogen replacement therapy and malignant transformation of endometriosis is documented, but malignant transformation following progestin therapy has not been reported previously. It appears that radical surgery is the only means of preventing malignant transformation of the lesion in patients with endometriosis. PMID- 11928866 TI - Pigmented (melanotic) neurofibroma. Report of an unusual case with immunohistochemical, ultrastructural and cytogenetic analyses. AB - In the spectrum of neurofibromas, pigmented tumors are rare variants usually showing only faint, macroscopically obvious pigmentation. We report a case of a huge pigmented neurofibroma with extended, macroscopically striking pigmentation in a patient with stigmata of neurofibromatosis. The immunohistochemical and ultrastructural findings support a melanotic line of differentiation besides schwann cell differentiation and indicate a phenotypic neoplastic spectrum between tumorous schwann cells and melanocytes. Using comparative genomic hybridization, striking chromosomal aberrations were not detected. High level amplifications of the known chromosomal regions, including genes of major enzymes responsible for melanin synthesis, appear to be unlikely. However, smaller chromosomal defects might have been overlooked by the limited resolution of this screening method. Therefore, other mechanisms up-regulating melanogenesis, such as mutations in regulatory genes, have to be considered. PMID- 11928867 TI - Intracellular distribution of intermediate filaments in vimentin-positive gastric carcinomas: confocal laser scanning microscopy using formalin-fixed paraffin embedded specimens. AB - Intermediate filaments are known as cytoskeletal elements. Recently, additional vimentin expression has been reported in some carcinomas; however, the function of such expression remains unclear. We studied the intracellular distribution of low-molecular weight cytokeratin and vimentin by immunohistochemistry in 17 vimentin-positive gastric carcinomas using confocal laser scanning microscopy. All materials were formalin-fixed and paraffin-embedded. Low-molecular weight cytokeratin expression showed a membranous pattern with a prominent deposition just below the cytoplasmic membrane in both tubular and solid components of the carcinomas. This unique membranous deposition was frequently absent in diffuse components. On the other hand, vimentin expression showed a fibrillary pattern in all components and also showed a unique basal distribution in the tubular components. We also recognized an aggregate pattern of the intermediate filament expression in diffuse components. We conclude that the significance of vimentin expression in carcinoma cells cannot be explained as a simple substitution for low-molecular weight cytokeratin because the distribution of vimentin and low molecular weight cytokeratin is different. PMID- 11928868 TI - Immunohistochemical detection of pS2 protein and heat shock protein-70 in pancreatic adenocarcinomas. Relationship with disease extent and patient survival. AB - We investigated pS2 and HSP-70 protein expression in 36 pancreatic adenocarcinomas for their effect on disease extent and patient outcome. The cases were reviewed, histologically diagnosed, typed, graded, and staged. Lymphatic vessel, blood vessel and perineural invasion as well as lymph node, resection margin and adjacent organ involvements were re-evaluated. The standard streptavidin biotin immunperoxidase method was used for immunostaining with pS2 and HSP-70 antibodies. Cytoplasmic staining with both antibodies was scored semiquantitatively. The scores were compared with histopathological prognostic parameters using statistical methods. Standard prognostic parameters and staining scores were tested by survival analysis in terms of their effect on survival. All the tumors showed a positive cytoplasmic reaction with HSP-70 antibody. Seventy seven percent of the tumors showed positive cytoplasmic staining with pS2 antibody (22.2% +, 13.9% ++ and 41.7% +++). There was a statistically significant difference between HSP-70 staining scores with N status and final stages of the tumors (Chi-square, p = 0.03 and p = 0.026, respectively), while neither direct nor inverse correlation was detected for both parameters. PS2 staining scores showed no statistically significant relationship with tumor grade T, M status, perineural invasion, lymph and blood vessel invasion. In tumors with extensive staining with pS2, tumor stage tended to be low (Chi square, p = 0.024, Kendall Tau-b, r: -0.336, p = 0.036). There was a statistically significant difference and inverse correlation between tumors with extensive pS2 staining and tumors with less intense staining in terms of lymph node metastasis (Chi-square, p = 0.041, Kendall Tau: p = 0.024, r = -0,373). In the R0 resection group, in univariate analysis, we found that with higher scores of HSP-70 staining, the prognosis of the patient tended to improve. (Cox regression, p = 0.013). In multivariate analysis, HSP-70 expression was found to be an independent prognostic factor. We found no relationship between pS2 staining and patient survival. PMID- 11928869 TI - Apoptotic bodies as a morphological feature in serous ovarian carcinoma: correlation with nuclear grade, Ki-67 and mitotic indices. AB - This study evaluated the relation of apoptosis with mitotic activity, Ki-67 indices, and nuclear and architectural grades in serous ovarian carcinoma. Apoptotic body (ABI) and mitotic indices (MI) were obtained by examining hematoxylin and eosin-stained sections from 35 serous ovarian carcinomas for apoptotic bodies and mitotic figures. ABI and MI were reported as the number of apoptotic bodies, and mitotic figures and immunostaining for Ki-67, respectively, as positive cells in 1000 cells. Nuclear grade was determined as grade 1 [n = 11], grade 2 [n = 13], and grade 3 [n = 11] according to recently defined criteria. There was a significant correlation between Ki-67 indices and ABI (p < 0.0001), Ki-67 and MI (p < 0.0001), as well as between MI and ABI (p < 0.0001). ABI increased with nuclear grade (p < 0.0001) and the type of the histological differentiation pattern (from glandular to papillary and solid architectural patterns) (p < 0.0001). Apoptosis, quantitated by ABI, is positively correlated with proliferation, thereby constituting a factor in the regulation of tumor growth of serous ovarian carcinomas. The positive correlation between ABI and increasing nuclear grade, mitotic activity, and architectural growth pattern may indicate that apoptotic bodies are another variable for grading serous ovarian carcinomas. PMID- 11928871 TI - Expression of p53 protein and tumor angiogenesis as prognostic factors in nasopharyngeal carcinoma patients. AB - The objective of this study was to evaluate the possible prognostic significance of p53 protein overexpression and tumor angiogenesis (TA) in nasopharyngeal carcinoma (NPC) patients, together with other clinicopathological variables. Forty-two NPC patients were evaluated in relation to survival. Nuclear p53 overexpression in neoplastic and endothelial cells was detected by immunohistochemistry (IHC) with the monoclonal antibody DO-7 and the polyclonal antibody against factor VIII-related antigen, respectively. Thereafter, we evaluated p53 cases in order to determine their nuclear immunoreactivity from negative (-) to positive (+, ++, +++). In addition, microvessels were counted in the most active areas of tumor neovascularization or hotspots using an image computer analyzer (MicroImage). A Cox multiple regression survival analysis was used to determine the best prognostic indicators in NPC patients. As a result, tumor microvessel count, considered as a continuous variable, was the most important independent prognostic indicator in relation to survival (p = 0.0273), with a relative risk of death of 2,4399 [95% confidence interval = 1.1051 ; 5.3871] associated with the highest microvessel counts. Moreover, the only clinicopathological variable that demonstrated prognostic value in a Cox multiple regression survival analysis was histological type (p = 0.05). In addition, we did not observe any statistical association between intratumoral microvessel density (IMD), clinicopathological variables and p53 protein expression. PMID- 11928870 TI - Assessment of transcriptional gene activity in situ by application of HOPE-fixed, paraffin-embedded tissues. AB - We report the use of HOPE-fixation (HOPE = Hepes-Glutamic acid buffer mediated Organic solvent Protection Effect) for specimens utilized for in situ hybridization targeting mRNA. For this purpose, an optimized protocol was developed and repeatedly tested on HOPE-fixed lung specimens. We observed that neither pretreatment, permeabilizing the cells, nor prehybridization is necessary to generate signals. After deparaffinizing, the random primed digoxigenin-labeled probes are directly hybridized together with yeast tRNA for blocking unspecific signals. Detection was performed using anti digoxigenin antibodies conjugated with alkaline phosphatase and new-fuchsine or NBT/BCIP as substrates. The results were verified by RT-PCR and adequate negative controls. Signals for human surfactant protein-A and interferon-gamma-inducible protein-10 developed rapidly within 10 min, accompanied by high signal intensities comparable to those observed in immunohistochemistry. Signal enhancement by biotinyl-tyramide, although giving suitable results as well, did not lead to higher signal intensities, and thus was not necessary in conjunction with the probes tested so far. These experiments were performed with material stored under appropriate conditions (at +4 degrees C) up to five years. To sum up, these initial results, obtained with the novel HOPE-fixative, are promising as regards the enhancement of the capabilities of in situ hybridization in the future. PMID- 11928872 TI - Drug use patterns among high school athletes and nonathletes. AB - This study examined drug use patterns and perceptions of drug intervention programs among adolescent interscholastic athletes and nonathletes. In particular, it explored the issue of whether participation in high school athletics is related to a healthier lifestyle and decreased use of recreational drugs and ergogenic aids. One thousand five hundred fifteen Massachusetts high school students completed a 150-item survey that assessed illicit and nonillicit substance use. Chi-square analyses revealed that athletes were significantly less likely to use cocaine and psychedelics, and were less likely to smoke cigarettes, compared with nonathletes. Conversely, nonathletes were less likely to use creatine than were athletes. There was no difference in the use of anabolic steroids and androstenedione between athletes and nonathletes. Descriptive analyses appear to indicate that drug interventions for athletes are falling short of their objectives. This study suggests that athletes have a healthier lifestyle and that the efficacy of intervention programs must be further examined. PMID- 11928874 TI - Family environment as a predictor of adolescent delinquency. AB - Efforts to decrease delinquency have led many researchers to study the underlying factors that lead youth to engage in delinquent acts. Webber (1997) has suggested that studies of delinquency can be categorized into those positing societal, individual, and/or family etiologies. The current research investigated the latter by assessing the relationship between family environment and delinquency. One hundred twenty-seven 9th-grade students completed the Family Adaptability and Cohesion Evaluation Scales (FACES III; Olson, Portner, & Lavee, 1985) and a modified version of Elliot and Ageton's (1980) Self-Report Delinquency Scale. Results indicated that the relationship between the two scales was moderated by family status (traditional versus nontraditional). The cohesiveness of the family successfully predicted the frequency of delinquent acts for nontraditional families, but failed to predict delinquency for traditional families. Implications for counselors, educators, and parents are discussed. PMID- 11928873 TI - Shame and guilt and their relationship to positive expectations and anger expressiveness. AB - College students (174 females, 91 males) completed measures of shame, guilt, expectations for future success, and styles of anger expression. Significant gender differences were found in proneness for both shame and guilt, with young women exhibiting a greater propensity for shame and guilt than young men. For both females and males, however, shame-proneness was positively related to expressions of inward anger. Among males and females, guilt-proness was negatively related to outward anger, but positively related to anger control. For females, guilt-proness was also negatively related to expectations for future success. Multiple regression analyses indicated that for male and female late adolescents, the best positive predictor of shame-proneness was inward anger. Gender differences emerged in predicting guilt-proneness; greater anger control, lower outward anger, and lower expectations for future success significantly predicted this variable among females. PMID- 11928875 TI - Gender, religion, and adolescent patterns of self-disclosure in the divided society of Northern Ireland. AB - Adolescence is a period when levels of self-disclosure are often lowest. While studies have revealed a clear preference for female targets of disclosure, little research has been carried out on the effects of religion upon disclosure. The impact of religion was of importance in this investigation, given that it was conducted in Northern Ireland, where religion affects almost every aspect of social life. The aim was to ascertain the effects of gender and religious affiliation on adolescent disclosure to friends and strangers. Results revealed that while females were significantly higher disclosers than were males, religion per se did not play a key role. This suggests that even in a highly polarized society, gender is the central determinant of disclosure and is even more important than political identity. The implications of these findings are discussed, particularly with regard to the difficulty young males have in terms of revealing personal information. PMID- 11928876 TI - Assimilation and ethnic boundaries: Israeli students' altitudes toward soviet immigrants. AB - The attitudes of Israeli students toward new immigrants from the former Soviet Union were examined against the theoretical framework of the sociology of ethnicity and sociology of education. The main research question explored was which ethnic relationship model (assimilation, pluralism, or separation) best described the attitudes and self-reported behavior of Israeli youth toward the new immigrants. Israeli students' changing attitudes toward their immigrant counterparts were also investigated, as well as actual Israeli educational practices (not only declared policies) regarding Soviet immigrants. The study was conducted in grades 4-11 at schools in northern Israel. The results indicated positive attitudes and openness of relations by the Israelis toward the new immigrant students in the first year following their immigration. However, the attitudes were less positive in the second year. Although the Israeli students manifested positive attitudes toward immigration and to the immigrants themselves, these views were largely assimilatory; Israeli cultural capital was obviously dominant, and it was expected to be accepted as such by the newcomers. The findings point to Israeli assimilatory educational practice; immigrants are expected to blend in, abandoning their past heritage and culture. PMID- 11928877 TI - Conflict behaviors and their relationship to popularity. AB - This study examined conflict behaviors (self, other) among 127 Turkish college students. Differences in five conflict behaviors (forcing, avoiding, accommodating, compromising, and collaborating) were then explored in relation to popularity and unpopularity. Results indicated that the students engaged in more avoiding and compromising behaviors, while perceiving more forcing behavior in others. Further, the unpopular group was found to engage in more compromising behavior, and perceived more forcing behavior in others, as compared with the popular group. Constructive and destructive conflict strategies, and their implications for popularity, are discussed. PMID- 11928878 TI - Attention deficit hyperactivty disorder in Korean juvenile delinquents. AB - This study was conducted to identify attention deficit hyperactivity disorder (ADHD) in Korean juvenile delinquents. Intelligence tests (KEDI-WISC, K-WAIS), the Test of Variables of Attention (TOVA), the Teacher Report Form (TRF), the Youth Self-Report (YSR), and the Rosenberg Self-Esteem Scale were administered to 98 incarcerated Korean adolescents (the delinquent group) and 84 adolescent nondelinquents (the control group). The groups were compared, and significant differences were found for ADHD; 42.4% of the adolescents in the delinquent group were identified as having ADHD, in comparison to 11.9% of the adolescents in the control group. Delinquent adolescents and adolescents with ADHD were found to have lower IQ scores, poorer TOVA performance, more severe problem behaviors, and lower self-esteem than nondelinquent adolescents and adolescents without ADHD. Delinquent adolescents with ADHD consistently fared the worst on assessments of intelligence, TOVA performance, problem behaviors, and self-esteem. PMID- 11928879 TI - Tobacco use among high school athletes and nonatieletes: results of the 1997 youth risk behavior survey. AB - In the United States today, the use of tobacco has become an entrenched part of teenage culture. The present study used the 1997 Youth Risk Behavior Survey (YRBS), which collected data from a nationally representative sample of 16,262 students in public and private high schools, to compare the tobacco use patterns of athletes and nonathletes. The independent variable, athletic participation, differentiated between moderately involved (1 or 2 teams) and highly involved (3 or more teams) athletes. Frequency of cigarette and cigar smoking and smokeless tobacco use served as the operational measure of tobacco use. Age, race/ethnicity, parental education, and residence were controlled. Logistic regression was used to calculate odds ratios for female and male athletes and nonathletes for each of the tobacco use variables. It was found that both male and female athletes were less likely to have ever smoked regularly, the effect being stronger for more highly involved athletes of both genders. Cigar smoking was unrelated to athlete status. Both female and male athletes were more likely to have used smokeless tobacco, the effect being stronger for more highly involved athletes of both genders. The findings are discussed in terms of access to health information, performance considerations, social status factors, the salience of an athletic identity, and the influence of the athletic subculture on its members. PMID- 11928880 TI - Frontal lobe functioning in adolescents with attention deficit hyperactivity disorder. AB - A prominent hypothesis regarding the etiology of attention deficit hyperactivity disorder (ADHD) is that its presence and magnitude reflect frontal lobe dysfunction. Past tests of this hypothesis have been inconsistent. The present study examined frontal lobe functioning in adolescents with ADHD. A sample of 10 ADHD adolescents and 10 controls between the ages of 12 and 17 served as participants for the study. The control group was matched on age (within six months) and gender. This study aided in clarifying past contradictory studies by using clearly defined criteria to determine ADHD, a representative age range of participants, carefully selected tests, and a control task. The two measures that were administered to assess frontal lobe functioning were the Wisconsin Card Sorting Test and the Stroop Color and Word Test. The Purdue Pegboard was used as a control measure that did not assess frontal lobe functioning. The findings indicate that the ADHD group performed significantly worse on the following: color score, color/word score, and interference score of the Stroop Color and Word Test; percent of perseverative responses, percent of perseverative errors, and number of completed categories of the Wisconsin Card Sorting Test. No differences were found on the Purdue Pegboard task. PMID- 11928881 TI - Academic goals, achievement, and age at first sexual intercourse: longitudinal, bidirectional influences. AB - This study examined bidirectional relationships between age at first sexual intercourse and academic goals and achievement. It was hypothesized that lower educational goals and achievement would be associated with initiating sexual intercourse at a younger age, and that initiating sexual activity early would be associated with a decrease in subsequent academic achievement and goals. In longitudinal data spanning 11 years, evidence was found for bidirectional effects. One interpretation of these results is that adolescents with high educational goals and achievement delay having intercourse because of the perceived risks (e.g., pregnancy and sexually transmitted diseases may jeopardize their plans for the future). Conversely, adolescents who engage in sexual intercourse at young ages might undergo a change in attitudes, including reduced interest in academic achievement and goals. The specific educational variables most strongly related to adolescent sexual intercourse in this study differed substantially by race and gender. PMID- 11928882 TI - Early puberty in African-American girls: nutrition past and present. AB - This study focused on nutrition and the changes in food intake over time to explain the onset of early pubertal signs in African-American girls. The daily nutritional intake of two diametrically opposite generations of African-American females was compared. The first group included 20 girls between the ages of 5 and 7 and the second group included 28 women between the ages of 70 and 85. The older African-American women were asked to remember the meals they had when they were young. Results revealed significant differences in terms of both quantity and quality of food consumed. Specifically, the women reported eating two meals a day when they were young while the girls reported eating at least three meals. Also, for breakfast, the girls consumed significantly more milk and fruits than did the women. For lunch, the girls consumed significantly more bread and grains, vegetables, meat, and milk than did the women. For dinner, the women consumed significantly more bread and grains. Thus, significant differences between the nutrition of African-American girls about 60 to 70 years ago and those of today were revealed. Implications of these findings are discussed. PMID- 11928883 TI - Adolescents' academic expectations and achievement. AB - Previous research has indicated that adolescents' relationships with their mother influence their academic expectations and achievement. Substance use has also been found to have a strong influence on academic expectations and achievement. In the present study, 80 high school seniors from middle to upper socioeconomic status families completed questionnaires on behavioral and psychological aspects of adolescent life. Academic expectations were found to be highly correlated with academic achievement (r = .60). A stepwise regression analysis revealed that relationship with mother, academic achievement, and alcohol use accounted for 56% of the total variance in academic expectations. Stepwise regression on academic achievement revealed that cocaine use, marijuana use, and academic expectations accounted for 48% of the total variance. PMID- 11928884 TI - Family, personality, and social risk factors impacting the retention rates of first-year Hispanic and Anglo college students. AB - This study investigated familial and behavioral differences between Hispanic and Anglo-American first-year college students. Analyses of variance and chi-square analyses were used to test for ethnic differences in (a) risk factors associated with family dysfunction, family addictions, personality factors, and social experiences, (b) student problem behaviors, namely alcohol use, drug use, and eating disorders, (c) a one-year follow-up of student retention, and (d) specific factors related to retention at the one-year follow-up. The Hispanic freshmen appeared to demonstrate resilience despite a lack of mentoring and greater paternal addiction. However, the one-year follow-up suggested that Hispanic females were at particular risk for leaving school. To improve college retention rates, social policy should focus on Hispanic students' vulnerabilities and strengths in making important developmental transitions. PMID- 11928885 TI - Racial and gender differences in weight status and dietary practices among college students. AB - A questionnaire was used to assess nutritional intake, weight status, and dietary practices among 630 U.S. college students. Body mass index (BMI) was calculated using self-reported heights and weights, and nutritional status was assessed via 24-hour recall. Sixty-four percent of the students had acceptable BMI levels (between 19 and 25); 16% of African-American females and 15% of African-American males had BMIs indicating obesity (30 or above). Approximately 18% of the students consumed 5 servings per day of fruits and vegetables, 7% consumed 6 or more grain products, and 53% consumed 2 or more dairy products. Twenty-seven percent reported never/rarely eating fast foods. Recommendations are presented for the development of culturally appropriate health education/promotion programs in order to help college students meet Healthy People 2010 objectives. PMID- 11928886 TI - Combining effect size estimates in meta-analysis with repeated measures and independent-groups designs. AB - When a meta-analysis on results from experimental studies is conducted, differences in the study design must be taken into consideration. A method for combining results across independent-groups and repeated measures designs is described, and the conditions under which such an analysis is appropriate are discussed. Combining results across designs requires that (a) all effect sizes be transformed into a common metric, (b) effect sizes from each design estimate the same treatment effect, and (c) meta-analysis procedures use design-specific estimates of sampling variance to reflect the precision of the effect size estimates. PMID- 11928887 TI - The role of qualitative research in psychological journals. AB - The acceptance of qualitative research in 15 journals published and distributed by the American Psychological Association (APA) was investigated. This investigation included a PsycINFO search using the keyword qualitative, an analysis of 15 APA journals for frequency of qualitative publication, a content analysis of the journal descriptions, and the results of qualitative interviews with 10 of the chief editors of those journals. The results indicate that there exists a substantial amount of interest in the potential contribution of qualitative methods in major psychological journals, although this interest is not ubiquitous, well defined, or communicated. These findings highlight the need for APA to state its position regarding the applicability of qualitative methods in the study of psychology. PMID- 11928888 TI - On the practice of dichotomization of quantitative variables. AB - The authors examine the practice of dichotomization of quantitative measures, wherein relationships among variables are examined after 1 or more variables have been converted to dichotomous variables by splitting the sample at some point on the scale(s) of measurement. A common form of dichotomization is the median split, where the independent variable is split at the median to form high and low groups, which are then compared with respect to their means on the dependent variable. The consequences of dichotomization for measurement and statistical analyses are illustrated and discussed. The use of dichotomization in practice is described, and justifications that are offered for such usage are examined. The authors present the case that dichotomization is rarely defensible and often will yield misleading results. PMID- 11928889 TI - Revisiting field experimentation: field notes for the future. AB - Field experiments in the social sciences were increasingly used in the 20th century. This article briefly reviews some important lessons in design, analysis, and theory of field experiments emerging from that experience. Topics include the importance of ensuring that selection into experiments and assignment to conditions occurs properly, how to prevent and analyze attrition, the need to attend to power and effect size, how to measure and take partial treatment implementation into account in analyses, modern analyses of quasi-experimental and multilevel data, Rubin's model, and the role of internal and external validity. The article ends with observations on the computer revolution in methodology and statistics, convergences in theory and methods across disciplines, the need for an empirical program of methodological research, the key problem of selection bias, and the inevitability of increased specialization in field experimentation in the years to come. PMID- 11928890 TI - Multiphase mixed-effects models for repeated measures data. AB - Behavior that develops in phases may exhibit distinctively different rates of change in one time period than in others. In this article, a mixed-effects model for a response that displays identifiable regimes is reviewed. An interesting component of the model is the change point. In substantive terms, the change point is the time when development switches from one phase to another. In a mixed effects model, the change point can be a random coefficient. This possibility allows individuals to make the transition from one phase to another at different ages or after different lengths of time in treatment. Two examples are reviewed in detail, both of which can be estimated with software that is widely available. PMID- 11928891 TI - Principles and practice in reporting structural equation analyses. AB - Principles for reporting analyses using structural equation modeling are reviewed, with the goal of supplying readers with complete and accurate information. It is recommended that every report give a detailed justification of the model used, along with plausible alternatives and an account of identifiability. Nonnormality and missing data problems should also be addressed. A complete set of parameters and their standard errors is desirable, and it will often be convenient to supply the correlation matrix and discrepancies, as well as goodness-of-fit indices, so that readers can exercise independent critical judgment. A survey of fairly representative studies compares recent practice with the principles of reporting recommended here. PMID- 11928892 TI - A comparison of methods to test mediation and other intervening variable effects. AB - A Monte Carlo study compared 14 methods to test the statistical significance of the intervening variable effect. An intervening variable (mediator) transmits the effect of an independent variable to a dependent variable. The commonly used R. M. Baron and D. A. Kenny (1986) approach has low statistical power. Two methods based on the distribution of the product and 2 difference-in-coefficients methods have the most accurate Type I error rates and greatest statistical power except in 1 important case in which Type I error rates are too high. The best balance of Type I error and statistical power across all cases is the test of the joint significance of the two effects comprising the intervening variable effect. PMID- 11928893 TI - A special report I. Prion protein (PrP)--amyloid plaques in the transmissible spongiform encephalopathies, or prion diseases revisited. AB - We present a retrospective analysis of PrP-amyloid plaques encountered in CJD and GSS. In human TSEs (kuru, CJD and GSS) several PrP-immunopositive plaques and plaque-like deposits were detected. In kuru, plaques were typical "kuru" plaques- stellate structures deposited mostly in the granular- and Purkinje-cell layer of the cerebellum. Many smaller or larger clusters were visible but, in contrast to GSS, they never merged together to form multicentric plaques. In all cases of GSS, plaques were localised in the granular- and Purkinje-cell layer and the molecular cell layer. There were many different forms of plaques: from kuru plaques (unicentric stellate plaques) to clusters of unicentric plaques, which by merging eventually formed "multicentric plaques". The latter are the hallmark of this disease. By electron microscopy, several types of amyloid plaques, which corresponded to those seen by PrP immunohistochemistry, were observed. The first type, unicentric "kuru" plaque, consisted of stellate arrangements (stars or cores) of amyloid bundles emanating from a densely interwoven centre. Amyloid stars were surrounded by astrocytic processes and invaded by microglial cells but dystrophic neurites were only rarely seen. In contrast, multicentric plaques were often surrounded by dystrophic neurites. The rarest type of plaque were neuritic plaques. In 263K- and 22C-H scrapie-infected hamster brains, by light microscopy and semi-thin (1 microm) sections, discrete PrP-immunopositive plaques were observed in the subependymal region but not in the deep brain neuroparenchyma. These plaques were not discernible by routine H & E staining. Ultrastructurally, plaques were recognised as areas of low electron density containing haphazardly oriented fibrils and not as stellate compact structures typical of plaques in human cases of CJD and GSS. These plaques were located beneath the basal border of the ependymal cells and adjacent blood vessels. Occasional dystrophic neurites containing electron-dense inclusion bodies were seen within the plaque perimeter, which always remained PrP-negative. PMID- 11928894 TI - Quantitative evaluation of intranuclear inclusions in SSPE: correlation with disease duration. AB - Three types of intranuclear inclusions in neurones, oligodendrocytes and astrocytes were quantitatively evaluated by electron microscopy in the autopsy material derived from six cases of subacute sclerosing panencephalitis (SSPE) with different duration of disease. Viral nucleocapsids were found in neurones and oligodendrocytes with the highest incidence (about 38% of nuclei) in two acute cases (adolescent), whereas in two subacute cases only 10% of nuclei of these cells contained nucleocapsids. However, in one acute case (child) and one chronic case, no nucleocapsids were detected at all, despite very intensive study. Two other types of intranuclear inclusions--nuclear bodies (NBs) and granulofilamentous inclusions (GFs) were present in astrocytic nuclei in all cases. Nuclear bodies were found the most frequently (about 66%) in cases of a several-week-long duration, and their incidence decreased with the extended duration of the disease. In the case of a seven-year-long duration, about 31% of nuclei contained NBs. The incidence of certain types of NBs varied also in individual groups of cases, and the same applied to the occurrence of cellular nuclei with different numbers of NBs. Nuclear bodies types IVand V occurred with similar frequency, regardless of the disease duration. The highest incidence of nucleocapsids and NBs was accompanied by the highest (about 25%) frequency of GF in astrocytic nuclei. The incidence of the latter declined with the prolonged duration of the disease, and in the chronic case it was about 16 times lower than in acute cases. In some acute and subacute cases, GF occurred together with NBs. Astrocytic nuclei with both types of inclusions occurred with a similar frequency (about 1.6-1.8%). PMID- 11928895 TI - Expression of vascular endothelial growth factor (VEGF) in rat brain after subarachnoid haemorrhage and endothelin receptor blockage with BQ-123. AB - Cerebral vasospasm is one of the most severe complications of subarachnoid haemorrhage (SAH), leading to pathological changes in the vessel wall itself and in the nervous tissue, due to ischaemia of endothelial cells and neurones. Amongst the known substances inducing vasospasm, the most potent spasmogenic effect is exerted by endothelin-1 (ET1). The constriction of cerebral arteries and obliteration of capillaries highly stimulates the secretion of growth factors by endothelial cells and induces compensatory formation of collateral circulation in response to brain ischaemia. Expression of vascular endothelial growth factor (VEGF), the main factor responsible for angiogenesis and vascular permeability, was found to be increased in hypoxic cells (irrespective of the cause of hypoxia) as well as in neoplastic cells in the brain. The aim of the study was to determine whether chronic vasospasm and hypoxia of endothelial cells stimulate expression of VEGF, and whether blockage of the endothelin receptor ET(A) reduces this expression. The SAH was induced experimentally in male Wistar rats and the ET(A) receptor antagonist--BQ-123 was administered into the cisterna magna. After 48 hours the brain was removed and expression of VEGF studied immunohistochemically on paraffin sections. We found that hypoxia of endothelial cells, induced by chronic vasospasm after SAH, caused increased expression of VEGF in brain vessels and neurones of the cerebral hemispheres, brain stem and cerebellum. After administration of the endothelin receptor antagonist BQ-123, no changes in VEGF expression in the brain were found. PMID- 11928896 TI - Controversies about the function of dystrophin in muscle. AB - Dystrophin, a product of a gene located at the chromosome Xp21 locus, is a cytoskeletal protein expressed in skeletal, cardiac and smooth muscles, and in the brain, and is located on the inner site of the plasma membrane. Dystrophin in the skeletal muscles is absent or appears only in traces in Duchenne dystrophy, it is reduced with normal/changed molecular weight in Becker dystrophy and it is absent/reduced in mdx mice. It is supposed that dystrophin acts either as a structural scaffold that supports mechanical stress in sarcolemma, or participates in regulating intracellular Ca2+ level. There are also data indicating that dystrophin takes part in force and signal transduction processes, in the aggregation of neurotransmitter receptors, and prevents an excessive generation of reactive oxygen free radical species. The main hypotheses indicate that lack of structural support, an excessive influx of Ca2+ ions into the muscle cell, or a combination of both these mechanisms in dystrophin-deficient muscle fibres, is responsible for muscle pathology in progressive muscular dystrophy. There are arguments supporting these hypotheses. There are, however, also data indicating that the presented arguments are doubtful. Despite recent advances in the knowledge of the pathogenesis of muscular dystrophies and the advent of modern techniques, we are still very far away from understanding what is the real function of dystrophin in muscle. PMID- 11928897 TI - Are all new data referring to amyotrophic lateral sclerosis certain? Some doubts. AB - Progress in molecular investigations presents new data facilitating the recognition of pathogenic mechanisms of numerous nervous system diseases, among them amyotrophic lateral sclerosis (ALS). Molecular studies of ALS are mainly concentrated on genetic search, excitotoxicity and astrocytic participation, pathology of neurofilaments, apoptosis, trophic factors, and selective motoneurone degeneration. In literature part of the results of these investigations are presented as certainty but some doubts exist. Problems of the role of the superoxide dismutase (SOD-1) mutation gene and free radical scavenging and selective vulnerability and death of motor neurone cells in ALS are mentioned. PMID- 11928898 TI - Tropical spastic paraparesis. AB - Human T-cell lymphotropic virus type I (HTLV-I) is the cause of endemic tropical spastic paraparesis (TSP) or HTLV-I-associated myelopathy (HAM). Because TSP/HAM is not a fatal disease, the neuropathology of this disease, albeit relatively well understood, is based on the examination of just a few incidental cases. We summarise our experience with the neuropathology of tropical spastic paraparesis/HTLV-I associated myelopathy (TSP/HAM). We studied three cases of TSP/HAM from different parts of the world. We demonstrated peculiar lamellated structures, called "multilamellar bodies" (MLB). It is tempting to suggest that MLB may represent specific ultrastructural markers of TSP/HAM. The pathology of the anteriorand posterior horns was similar and comprised axonal degeneration, accompanied by extensive astrocytic gliosis. Lymphocytic infiltration, particularly observed as "cuffs" around blood vessels, was scattered among other cellular elements. Ultrastructurally, myelin sheaths were relatively well preserved, and some demyelinated but not remyelinated fibres were observed. Moreover, axons with abnormal accumulations of neurofilaments, suggestive of axonal degeneration, were detected. Several axons contained Hirano bodies. In many samples glial processes replaced most of the remaining neuropil. PMID- 11928899 TI - Ultrastructural study of mother and daughter muscle changes with mitochondrial encephalomyopathy. AB - We present the light and electron microscopy examinations of skeletal muscle biopsies from a 36-year-old mother and her 13-year-old daughter with mitochondrial encephalomyopathies. Clinical signs and symptoms suggesting mitochondrial disease, such as disseminated neurological symptoms, visual and hearing disturbances, mental disability, exercise intolerance, heart conduction disturbances, short stature, family history, were present in both patients. The mother's niece (8 years old) also died with progressive neurological disorder. CT showed cerebral and cerebellar atrophy in mother and multifocal subcortical atrophy in daughter. There was lactic acidosis in blood serum and cerebrospinal fluid in daughter. In the daughter's muscle a lot of fibres looked like ragged red fibres. Electron microscopic examination revealed the alterations of mitochondria in skeletal muscle of both patients that concerned the number, size, shape and the fine structural appearance of the mitochondria. The most characteristic mitochondrial abnormalities in daughter's muscles were paracrystalline inclusions in the intracristal space. In mother's muscles most of the mitochondria were markedly enlarged and they possessed aberrant configurations of cristae. The mitochondrial matrix contained sometimes spherical electron dense bodies different in size and vacuoles. Ring-shaped mitochondria were also observed. The most prominent ultrastructural feature, similarly as in daughter, was the occurrence of intramitochondrial highly ordered paracrystalline inclusions. PMID- 11928900 TI - Morphometric analysis of the cerebellar cortex capillaries in the course of experimental valproate encephalopathy and after chronic exposure to sodium valproate using transmission electron microscopy. AB - Morphometric analysis of the cerebellar cortex capillary cross-section area performed in experimental valproate encephalopathy using transmission electron microscopy showed that prolongation of VPA application resulted in more enhanced lumen narrowing manifested in gradual reduction in the mean value of the coefficient examined. After 6, 9 and 12 months of experiment this value was statistically different from that obtained in control subgroups, being respectively lower by approximately 22%, 48% and 65%. One month after terminating of chronic administration this value was close to the one found after 12 months of the study. Three months after the drug withdrawal the coefficient was higher by approximately 44% compared to the one after 12 months, which seemed to indicate an increase in capillary lumen patency. The morphometric analysis of the cerebellar cortex capillary cross-section area performed in the present study objectifies the results of qualitative ultrastructural investigations concerning the microcirculation of this CNS structure. PMID- 11928901 TI - Guillain-Barre syndrome in patient with Burkitt's lymphoma and type 2 diabetes mellitus. AB - Although peripheral neuropathies are commonly observed in patients with non Hodgkin's malignant lymphomas (NHML), Guillain-Barre syndrome (GBS) belongs to the occasional complications of lymphoproliferative disorders. It appears in less than 0.3 per cent of NHML. It is worthy of note that in the reported case there occurred three independent risk factors of peripheral neuropathy: Burkitt's lymphoma, chemotherapy and type 2 diabetes mellitus. Based on clinical course, EMG finding and neuropathological examination, in spite of normal cerebrospinal fluid protein content, GBS as a paraneoplastic disorder was diagnosed. It was assumed that chemotherapy and diabetes mellitus conduced to severe neuropathy. PMID- 11928902 TI - Failure of HA coating on a gritblasted acetabular cup: 155 patients followed for 7-10 years. AB - We report the outcome of 191 acetabular gritblasted titanium cups with a hemispherical design for press-fit insertion and coated with hydroxyapatite. The prosthesis was made of gritblasted titanium entirely coated with hydroxyapatite. 155 patients aged 15-78 years were operated on during the years 1991-1993 and followed for 7-10 years. During this period, 39 cups were revised because of mechanical loosening, a further 9 had radiolucent lines and 2 focal osteolysis. None of these 11 patients had clinical symptoms. Failure was associated with age, wear and radiolucency/osteolysis. At revision, we found that the soft tissues were discolored, and that most of the coating had disappeared. This design of hydroxyapatite-coated cups has a high rate of debonding and failure. PMID- 11928903 TI - Hip ganglion cyst associated with developmental dysplasia of hip in a child-a case report. PMID- 11928904 TI - Evaluation of four experimental osteomyelitis infection models by using precolonized implants and bacterial suspensions. AB - Staphylococcus aureus osteomyelitis, a major problem in orthopedic surgery, often involves biofilm bacteria adhering to implants and surrounding bone and tissues. The inadequacy of therapy or immunological surveillance has encouraged studies using animal models which simulate natural osteomyelitic infections, ensure the development of infections and avoid mortality. We evaluated 4 models for infection (8 animals/model) in rats, using stainless-steel implants in tibiae and a very adherent slime-producing bacterial strain. Each animal received: an implant containing a 12 h-biofilm with about 10(6) cfu (Model 1); an implant containing this biofilm and a suspension with about 10(4) cfu (Model 2): a sterile implant and a suspension with about 10(5) cfu (Model 3); or a sterile implant and a suspension with about 10(6) cfu (Model 4). 63 days after surgery we found 100% rat survival, colonization of bone by implant biofilm bacteria in some animals and local, but not systemic infections. Model 1 (but not Models 2-4) reproduced an infection in both, tibiae and implants, most reliably (in 100% of the animals). Model 3 was the least reliable (p < 0.01, 25% infected implants, 12% infected tibiae). PMID- 11928905 TI - Preoperative nutritional evaluation as a prognostic tool for wound healing. AB - In 170 patients given primary hip or knee joint prosthesis surgery, we determined the relation between variables connected with the preoperative nutritional state and delayed healing of the surgical wound. 46 patients had a lymphocyte count of less than 1500 cells/mm3 (normal 1500-3300 cell/mm3), 18 an albumin level of less than 3.5 g/dL (normal 3.5-5 gr/dL), and 35 patients had a transferrin level of less than 200 mg/dL (normal 200-400 mg/dL). We found an association between the preoperative lymphocyte count and delay in wound healing, whereas preoperative seru albumin and transferrin levels had no significant predictive value. In addition, a preoperative lymphocyte count of less than 1500 cells/mm3 was associated with a 3 times higher frequency of healing complications. PMID- 11928906 TI - Bone allografts pretreated with a bisphosphonate are not resorbed. AB - Bisphosphonates bind to bone surfaces and inactivate osteoclasts when they start to resorb the bone. Therefore, immersion of a bone graft in a bisphosphonate solution before implantation may protect it from resorption. We implanted frozen cancellous bone allografts into bilateral bone chambers for 6 weeks in 10 rats. One graft in each pair had been immersed in an alendronate solution (1 mg/mL) for 10 minutes, and then rinsed in saline. Controls underwent the same treatment with saline only. Results were evaluated with histomorphometry. Control grafts were almost entirely resorbed, but alendronate-treated grafts seemed intact. In the treated specimens, two thirds of the space behind the bone ingrowth frontier consisted of graft or host bone, but in the controls, only one fifth. Local graft treatment with a bisphosphonate before insertion seems to be risk-free, and may prevent mechanical graft failure due to resorption in patients. PMID- 11928907 TI - Reduction of instability-induced bone resorption using bisphosphonates: high doses are needed in rats. AB - Bone resorption associated with prosthetic loosening can be reduced by giving bisphosphonates since they bind to bone surfaces and inactivate osteoclasts when bisphosphonate-containing bone is resorbed. During loosening, an increase in osteoclastic activity can be triggered by mechanical instability, fluid pressure or wear particles. We used a rat model in which a titanium surface can be made to slide over a bone surface and cause instability-induced bone resorption. 111 rats were operated on with a plate implant and treated with alendronate or clodronate injections in different doses or saline controls. After 4 weeks of osseointegration, the plate was moved during 2 weeks and the findings evaluated with histomorphometry. The percentage of persisting bone-metal contact and the soft tissue area at the interface were measured to estimate bone loss. Low or intermediate doses of the bisphosphonates increased the ash weight of untraumatized bone, but did not inhibit resorption at the unstable interface. Only rats treated with the highest doses of alendronate or clodronate had more bone-metal contact than controls. Instability-induced bone resorption therefore seems to be reduced by bisphosphonates, but higher doses are needed to obtain this effect than to reduce bone resorption associated with normal remodeling of untraumatized bone. PMID- 11928908 TI - Osteogenic protein 1 device increases bone formation and bone graft resorption around cementless implants. AB - In each femoral condyle of 8 Labrador dogs, a non weight-bearing hydroxyapatite coated implant was inserted surrounded by a 3 mm gap. Each gap was filled with bone allograft or ProOsteon with or without OP-1 delivered in a bovine collagen type I carrier (OP-1 device). 300 microg OP-1 was used in the 0.75 cc gap surrounding the implant. After 3 weeks, the OP-1 device enhanced implant fixation by 800% (p <0.05) in the ProOsteon group, but OP-1 bad no significant effect on implant fixation in the allograft group. By adding the OP-1 device, the volume fraction of woven bone close to the implant increased from 12% to 20% (p < 0.05) in the bone allograft group and from 6% to 25% (p < 0.05) in the ProOsteon group. The volume fraction of bone allograft decreased from 29% to 9% (p < 0.05) in the OP-1 treated group versus 33% to 30% in the allograft group not treated with OP 1. No resorption of ProOsteen was found. In conclusion, OP-1 accelerates resorption of bone allograft and enhances new bone formation around cementless implants grafted with bone allograft or semisynthetic hydroxyapatite bone substitute. Our findings do not support the use of ProOsteon alone around cementless implants. PMID- 11928909 TI - Atrophy of the rotator cuff muscles and site of cuff tears. AB - We determined the relationship between the site of rotator cuff tears and atrophy of the cuff muscles. 28 shoulders (28 patients) had rotator cuff tears: 19 isolated tears of the supraspinatus tendon (isolated-tear group) and 9 combined tears of the supraspinatus and infraspinatus tendons (combined-tear group). The cross-sectional area of the subscapularis, supraspinatus, the infraspinatus and teres minor muscles in the coronal oblique MR images were measured before and after surgery. Although we found no difference in tear size, the cross-sectional areas of the muscles were smaller in the combined-tear group than in the isolated tear group. We conclude that atrophy of the supraspinatus and infraspinatus muscles also depends on the site of the tear. PMID- 11928910 TI - Strength and motion after hemiarthroplasty in displaced four-fragment fracture of the proximal humerus: 27 patients followed for 1-6 years. AB - We evaluated 27 patients with shoulder hemiarthroplasty after displaced four fragment fracture of the proximal humerus after mean 4 (1-6) years. Isometric strength measurements (Kintrex) and three-dimensional motion analysis (Elite System) were performed on the operated and unoperated shoulders. Clinical assessment was based on Constant's score and Neer's scoring system. The isometric strength of the operated and unoperated sides were 22 (SD 8.6) Nm and 24 (SD 5.9) Nm in abduction and 48 (SD 14) Nm and 65 (SD 21) Nm, respectively in adduction (the latter was statistically significant). Motion analyses at follow-up showed a mean reduction in glenohumeral movement. Increases in acceleration and deceleration of the acromion at the operated side were noted, indicating a change in glenohumeral rhythm during maximal abduction. The Constant score was 45 (SD 15) points with a significant reduction in the range of motion. 15 patients had some degree of heterotopic ossification. On the basis of our findings, the impaired function seems to be caused by reduced glenohumeral mobility rather than muscle strength. We also found a better outcome after early than late hemiarthroplasty. PMID- 11928912 TI - The ScanHip total hip arthroplasty: radiographic assessment of 72 hips after 10 years. AB - We analyzed the radiographic and clinical outcome of the ScanHip total hip arthroplasty in 70 patients after 10 years. The Swedish National Hip Register, in which the end-point of the survival analysis is defined as revisions, reported a 10-year survival rate of 94% with the ScanHip, but in the present series 13% of the femoral stems and 29% of the sockets met the criteria for aseptic loosening. Focal osteolysis was found around 8 sockets (11%) and 23 stems (32%) and had occurred significantly oftener around loose sockets, but not around femoral stems. Linear polyethylene wear was significantly increased in loose sockets, but no relationship was noted between polyethylene wear and stem loosening or was there a correlation between clinical symptoms or patients' satisfaction and component loosening. Therefore precise serial long-term radiographic follow-up is the only satisfactory method for detecting aseptic loosening of total hip arthroplasty. It gives the surgeon more detailed information about each case than survival analysis alone. PMID- 11928911 TI - Surgical treatment of unstable fractures of the distal clavicle: a comparative study of Kirschner wire and clavicular hook plate fixation. AB - During 1988-1999 39 unstable fractures of the distal clavicle (Neer 2) were operated on in Oulu University Hospital. Kirschner wire (K-wire) fixation was used in 22 cases and a clavicular hook plate in 17. Shoulder symptoms and function were assessed using self-administered questionnaires devised by L'Insalata et al. (1997), and Constant scoring. Mean follow-up was 6.2 years in the K-wire fixation group and 2.0 years in the clavicular hook plate one. The mean L'Insalata scores were 91 in both groups (92% and 93% of the contralateral side) and the mean Constant scores 84 (95% and 90 (96%) for K-wire fixation and the clavicular hook plate, respectively. Complications commonly occurred with K wires, which migrated in 12 cases, resulting in loss of reduction in 7 and infection in 3, and 2 cases of non-union. In the clavicular hook plate group, there was 1 complication, a fracture of the clavicle, and 2 cases of non-union. We conclude that shoulder symptoms were reduced and function restored to an adequate level by both methods, but complications were unacceptably frequent when K-wires were used. The clavicular hook plate was better in this respect and it is therefore recommended. PMID- 11928913 TI - Changes in the production and the effect of nitric oxide with aging in articular cartilage: an experimental study in rabbits. AB - We studied the production and the effect of nitric oxide (NO) in articular cartilage from rabbits of various ages. 40 New Zealand white rabbits in 4 age groups (1 month, 6 months, 1 year, and 3 years of age with 10 rabbits in each group) were used. Basal and induced levels of NO were lower in cultured chondrocytes from older rabbits. Exogenous NO administration suppressed the proliferative activity of chondrocytes to a greater degree in younger rabbits. Immunohistochemistry showed that older rabbits had fewer eNOS positive chondrocytes. Our findings imply that the age-related change in NO in articular cartilage does not have a relevance to increased NO production in osteoarthrosis. PMID- 11928914 TI - Fixation with poly-L-lactic acid screws in hip osteotomy: 68 hips followed for 18 46 months. AB - This study evaluated 68 consecutive hip osteotomies in 61 patients using absorbable poly-L-lactic acid screws for fixation. 47 hips underwent a rotational acetabular osteotomy, 17 hips Chiari's pelvic osteotomy, and 4 hips transtrochanteric rotational osteotomy. Cortical screws were used to transfix the osteotomized acetabulum, and cancellous screws to reattach the intraoperatively osteotomized greater trochanter. The average age at surgery was 35 (12-49) years. The mean duration of follow-up was 32 (18-46) months. All the osteotomized acetabulums united well, but 4 of 54 trochanteric osteotomies failed to unite. PMID- 11928915 TI - Components of laxity in interference fit fixation of quadrupled hamstring grafts. AB - We tested in vitro 15 Poly-L-lactic acid and 14 titanium interference screws in male human cadaver tibia-hamstring constructs. Elongation of the graft, displacement of the screw in the tibial tunnel, and slippage at the graft/screw interface were measured as outcomes with an infrared optical system. All constructs failed by slippage of the graft past the interference screw. On average, graft slippage accounted for 92% of total construct laxity at 5mm total construct elongation, whereas permanent stretching of the midsubstance of the graft accounted for only 8%. Movement of the interference screw in the tibial canal was minimal, averaging 0.01 (0-0.5) mm at construct failure. Inadequacy of isolated tibial interference screw fixation of soft tissue grafts may be overcome if early failure at the graft/screw interface is prevented. PMID- 11928916 TI - Supracondylar femur fracture fixation: mechanical comparison of the 95 degrees condylar side plate and screw versus 95 degrees angled blade plate. AB - The best way to stabilize supracondylar femur fractures remains debatable. Previous studies have compared internal fixation to intramedullary fixation, but none have compared the stiffness characteristics and strength of the 95 degrees angled blade plate (ABP) with the 95 degrees condylar side plate and screw (DCS). 14 synthetic femora were cut in half and the proximal pole of the distal fragment was made secure. A 1 cm gap was made parallel to the femoral condylar weight bearing surface to create an extraarticular supracondylar femur fracture (OTA 33 A3). 7 femora were stabilized with an ABP and 7 with a DCS. Using an MTS compression/torsion servohydraulic testing machine, each femur was tested in 7 modes of loading: (1) axial compression; (2) anterior compression; (3) posterior compression; (4) medial compression; (5) lateral compression; (6) torsion in external rotation; and (7) torsion in internal rotation. The stiffness of the construct in each mode, the "maximum load in axial compression", and the fatigue characteristics in axial compression were measured. The DCS showed a statistically significant greater stiffness in axial compression and average maximal load than the ABP. The fatigue tests revealed no evidence of permanent deformation or loosening of either construct. PMID- 11928917 TI - Evidence for a neuropathic contribution to the development of spontaneous knee osteoarthrosis in a mouse model. AB - Previous work has shown a progressive, age-related loss of knee joint innervation in the C57BL6Nia mouse. We did three experiments to describe further the loss and determine whether it might contribute to the development of knee osteoarthrosis in this model. Immunocytochemistry showed that the percentage of neurons expressing substance P and calcitonin gene-related peptide increased with age, indicating a relatively selective loss of mechanoreceptors. Histological examination of knee joints of mice at various ages showed that loss of joint innervation always preceded histological changes of cartilage degeneration. The mice usually developed a mild form of osteoarthrosis, but surgical ablation of joint innervation caused the development of severe patellofemoral osteoarthrosis. The findings are consistent with the hypothesis that an age-related loss of joint innervation may contribute to the development of osteoarthrosis. PMID- 11928918 TI - Rotationplasty--quality of life after 10 years in 22 patients. AB - We assessed the long-term results in 22 patients with rotationplasty after resection of high-grade malignant bone tumors. We used established methods (QLQ-C 30; FLZ) to evaluate the quality of life, diplomas, life-contentment, occupational situation and marriage status. After 10-year follow-up, we found no reduction in psychosocial adaptation, and life contentment was about the same as in healthy persons. We therefore recommend rotationplasty instead of amputation whenever conventional limb salvage is impossible. PMID- 11928919 TI - Exsanguination of lower limbs in healthy male subjects. AB - Gamma camera technique was used to assess the effectiveness of various exsanguination methods in 12 healthy male volunteers given an autologous injection of 99mTc-labeled erythrocytes. The methods used included elevation alone, Esmarch bandage, gauze bandage, and the Pomidor roll-cuff. The median times spent on use of these methods were: Esmarch 85 sec, gauze 104 sec, and Pomidor roll-cuff 18 sec. The various exsanguination methods caused a median percentage reduction in regional blood volume of the lower limbs: elevation 1/2 minute 45%, 1 minute 45%, 2 minutes 42%, 4 minutes 44%, 6 minutes 43%, 10 minutes 44%, Esmarch bandage 64%, gauze bandage 62%, and Pomidor roll-cuff 61%. No statistically significant differences were found between the elevation procedures. The external methods were more effective than elevation alone (p < 0.001). PMID- 11928920 TI - Reduced lengthening index by use of bifocal osteotomy in the tibia: comparison of monofocal and bifocal procedures with the Ilizarov external fixator. AB - We lengthened 63 tibial segments in 49 patients, performing a monofocal osteotomy on 31 segments and a bifocal osteotomy on 32 segments. In the monofocal osteotomy group, the average lengthening was 4.8 (2.4-7.1) cm with a lengthening index (LI) of 1.7 (0.9-4.4) months/cm. In the bifocal osteotomy group, the average lengthening was 7.4 (4.9-10.0) cm and with a LI of 1.0 (0.7-2.1) months/cm. The difference in LI between the groups was statistically significant, but more major complications, like persistent ankle joint contracture and pseudarthrosis in the callotasis lengthening zone, occurred with bifocal lengthening. PMID- 11928921 TI - Tibio-calcaneo-naviculo-cuboidale arthrodesis: 6 patients followed for 1-8 years. AB - As a salvage procedure for patients with irreparable damage to the peritalar anatomy, we used a new method, the Ilizarov ring fixator, to stabilize a tibio calcaneo-naviculo-cuboideal (TCNC) arthrodesis after resecting the talus in 6 patients with an average follow-up of 3 years. The tibia was lengthened by callus distraction, mean 6 (3.5-10) cm. Patients with exacerbation of a chronic talar osteomyelitis, symptomatic posttraumatic changes, and those with extremely unstable club feet and talar deformity preventing an adequate reduction of the joint, were successfully treated with the TCNC-fusion. The method permits early weight bearing, while providing stability and compression of the fusion, thereby avoiding further loss of bone stock. PMID- 11928922 TI - A review of canine pseudocyesis. AB - The purpose of this article is to review the most relevant features of the physiology, clinical signs, diagnosis, treatment and prevention of canine pseudocyesis (PSC). This is a physiological syndrome, characterized by clinical signs such as: nesting, weight gain, mammary enlargement, lactation and maternal behaviour, which appears in non-pregnant bitches at the end of metaoestrus. PSC is a frequent finding in domestic dogs. Although it is generally admitted that prolactin (PRL) plays a central role in the appearance of PSC, its precise aetiophysiology is not completely understood yet. A number of clinical studies suggest that at some point of metaoestrus circulating PRL levels rise in overtly pseudopregnant bitches. Individual differences in sensitivity to PRL as well as the existence of molecular variants of canine PRL with different bioactivity versus immunoreactivity ratios may help clarify the aetiopathology of PSC. Diagnosis of PSC is based on the presence of typical clinical signs in metaoestrous non-pregnant bitches. Considering that PSC is a self limiting physiological state, mild cases usually need no treatment. Discouraging maternal behaviour and sometimes fitting Elizabethan collars to prevent licking of the mammary glands may suffice in these cases. Sex steroids (oestrogens, progestins and androgens) have been traditionally used to treat PSC but the side-effects usually outweigh the benefits of these medications. Inhibition of PRL release by ergot derivatives [bromocriptine (10-100 microg/kg per day for 10-14 days], cabergoline (5 microg/kg per day during 5-10 days), metergoline (0.2 mg/kg per day during 8-10 days) has proved to be effective for the treatment of canine PSC. Although some of these ergot derivatives present some untoward side-effects, they are transient and can usually be managed. Predisposed bitches not intended for breeding should be spayed as ovariectomy is the only permanent preventive measure. PMID- 11928923 TI - Gross morphometry of the bovine placentome during gestation. AB - Forty-seven pregnant uteri were collected from an abattoir at eight chronological stages of gestation. Assessments were made of placentome number, weight, length shape and ratio of foetal to maternal tissue by weight. The study showed that there was a great variation in number, size, weight and shape of placentomes. There was no significant change in the total number of placentomes during gestation. Both mean weight and length increased significantly during gestation, but there was a significant change in the relationship between the two as gestation progressed. There was no correlation between the number of placentomes and the mean weight or length of the placentomes. The development of the placenta in the non-pregnant horn was significantly different from that of the pregnant horn, with fewer, smaller, lighter placentomes. However, there was no evidence that placentome development in the non-pregnant horn was affected by placentome development in the pregnant horn. The proportion of the placentome that was maternal tissue increased during gestation, but there was no significant effect of placentome number, or individual placentome weight or length on this proportion. Two placentome shapes were identified, but no effect on foeto maternal proportion or relationship between shape and total placentome number or weight was identified. PMID- 11928924 TI - Litter size and vagina-cervix catheter penetration length in gilts. AB - As in other species, the reproductive tract in pigs increases in size with age and body weight, and the development of the reproductive tract depends on a balance between development of the pituitary-ovarian axis and the influence of metabolic hormones. Two experiments were conducted in prepubertal Duroc gilts, 150-180 days of age, to determine whether litter size is related to vaginal cervix catheter penetration length during insemination. In experiment 1, oestrus was induced in 452 gilts with a combined dose of 400 IU Pregnant Mare Serum Gonadotrophine (PMSG) + 200 IU human chorionic gonadotropin (hCG). The gilts were classified into three catheter penetration length groups: Ih, < or = 21 cm; IIh, > 21 and < 28 cm, IIIh, > 28 cm. The litter size was lowest in group Ih (7.35 +/- 0.15) compared with groups IIh (7.81 +/- 0.12; p < 0.05) and IIIh (10.0 +/- 0.36; p < 0.001). In experiment 2, first oestrus was induced in 162 gilts by boar exposure. The gilts were classified into three catheter penetration length groups at insemination during their second oestrus: In, < or = 24 cm; IIn, > 24 and < 26 cm; IIIn, > 26 cm. As in experiment 1, the litter size was lowest in the group with the shortest catheter penetration length (8.32 +/- 0.19). The litter size was not different among gilts of groups IIn and IIIn (8.84 +/- 0.35 and 9.56 +/- 0.46, respectively), but litter size was lower (p < 0.05) in group In than in group IIn. Based on the combined data from both experiments, the correlation between the catheter penetration length and total number of piglets born was expressed as: y =5.346 +/- 0.104x; r = 0.361 (p < 0.05). Fertility rate was not different among the groups of gilts induced into oestrus by hormone treatment or inseminated in the second oestrus; however, the total fertility rate of boar exposed gilts was higher (p < 0.0001) than PMSG/hCG treated animals. Thus, it is possible to conclude that litter size at first farrowing is associated with vaginal-cervix catheter penetration length during insemination of the gilt. PMID- 11928925 TI - Effect of oestrous synchronization with estradiol 17beta and progesterone on follicular wave dynamics in dairy heifers. AB - An experiment was designed to evaluate the effects of estradiol-17beta (E17beta) on follicular wave dynamics and ovulatory response in Holstein heifers receiving either a progestogen ear-implant (Crestar; Intervet International b.v. Boxmeer, The Netherlands) or an intravaginal progesterone-releasing device [controlled internal drug release-bovine device (Eazibreed, CIDR-B; Bodinco BV, Alkmaar, The Netherlands)]. For comparison, another group of heifers was also synchronized using Crestar plus an injection of estradiol valerate (EV) and norgestomet as recommended by the pharmaceutical company. Twenty 20-22-month-old cycling Holstein heifers were allocated to one of the following treatment groups at random stages of the oestrous cycle: (I) simultaneous insertion of Crestar and intramuscular injection of 3 mg norgestomet and 5 mg EV (Crestar 9 + EV 9); (II) simultaneous insertion of Crestar and intramuscular injection of 5 mg E17beta (Crestar 9 + E17beta 9); (III) insertion of Crestar followed 2 days later by intramuscular injection of 5 mg E17beta (Crestar 9 + E17beta 7); or (IV) insertion of CIDR-B device followed 2 days later by intramuscular injection of 5 mg E17beta (CIDR 9 + E17beta 7). The CIDR-B or Crestar implants were removed after 9 days and all heifers received 500 microg Cloprostenol (Estrumate, Pitman Moore Nederland BV, Houten. The Netherlands). Ovarian ultrasonographic examinations were performed once daily during the synchronization period using a B-mode scanner equipped with a 7.5 MHz linear-array transrectal transducer. In addition, heifers were scanned every 12 h after implant/device withdrawal until 3 days after ovulation in order to monitor follicular activity, detect ovulation and subsequent early luteal formation. Detection of oestrus was performed every 6 h for 4 days after device/implant removal. Oestrus was observed 24-32 h before ovulation in all heifers. The mean hours interval from treatment withdrawal to ovulation was not significantly different (84.0 +/- 16.5, 77.6 +/- 4.1, 73.6 +/- 4.1 and 64.0 +/- 4.4 h for treatments I, II, III and IV, respectively, p > 0.1). However, the variance for heifers treated with EV + norgestomet was significantly larger (Levene's Test; p < 0.01) than those treated with E17beta. All E17beta treatments resulted in dominant follicle suppression and a new wave emerged 4.1 days after treatment compared with 6.6 days for the EV + norgestomet treatment (p < 0.05). The time from emergence of the new ovulatory wave to ovulation was longer for the new wave that emerged after E17beta treatment (9.2 +/- 0.3 days) than after EV + norgestomet treatment (6.9 +/- 0.4 days; p < 0.05). The results of this study suggest that the four treatments used were effective in inducing synchronous behavioural oestrus and ovulation. However, a higher degree of oestrus and ovulation synchrony was observed in heifers treated with E17beta than in heifers treated with EV + norgestomet. Synchronization treatments with exogenous E17beta or EV + norgestomet at the time of progestin device insertion (Crestar or CIDR-B) or 2 days later in heifers can regulate a different emergence pattern of ovarian follicular development in randomly cyclic heifers. The E17beta was effective in inducing follicular suppression and resulted in the consistent emergence of a new follicular wave. PMID- 11928927 TI - Improvement of the culture conditions for in vitro production of cattle embryos in a portable CO2 incubator. AB - The effects of different concentrations of growth hormone (GH) on in vitro maturation (IVM), fertilization (IVF) and culture (IVC) of bovine oocyte/embryos in CR1aa or CR2aa media using a simple CO2 incubator were investigated. The IVM/IVF/IVC of oocytes were carried out in the presence of 0, 50, 100 and 200 ng/ml GH in the medium. The proportion of metaphase II oocytes was significantly higher (p < 0.05) in 200 ng/ml compared with 0 ng/ml GH in CR1aa medium (59 versus 85%, respectively), but this effect was not observed under CR2aa. Higher concentrations of GH yielded lower rates of unfertilized ova and thus superior cleavage rates (36.5 +/- 0.2 and 63.5 +/- 2.0% versus 17.5 +/- 0.2 and 82.5 +/- 1.5% or 40.4 +/- 0.6 and 59.6 +/- 1.4% versus 16.6 +/- 1.2 and 83.4 +/- 6.2% for 0 and 200 ng/ml GH in portable or ordinary incubator, respectively) in CR1aa. This dose-dependent effect was also observed in the percentages of transferable embryos, although not statistically different (17.2 +/- 1.7 versus 27.3 +/- 1.8% and 16.6 +/- 3.1 versus 26.0 +/- 1.4%, for 0 versus 200 ng/ml GH in portable and ordinary incubator, respectively). In contrast to the CR1aa, different concentrations of GH in CR2aa medium did not increase either fertilization or cleavage rates. In fact, higher concentrations of GH in this medium negatively affected the rate of transferable embryos. Hence, percentages of transferable embryos obtained in the portable incubator under 0 or 50 ng/ml GH were higher (p < 0.05) compared with those obtained in 100 or 200 ng/ml GH (35.4 +/- 5.7 or 40.5 +/- 5.4% versus 22.4 +/- 2.4 or 15.5 +/- 2.1%, respectively). There was however, no significant difference in the rate of transferable embryos in an ordinary incubator employing CR2aa medium, but the trend was more or less similar to that observed in the portable incubator. Despite the fact that relatively fewer oocytes were employed for the culture in the ordinary incubator, overall results observed employing the simple portable CO2 incubator were within the range of those obtained in an ordinary incubator: implying that the simple portable incubator can effectively be employed for the in vitro production of bovine embryos under field conditions. PMID- 11928926 TI - First confirmed case of Neospora caninum-associated abortion outbreak in Portugal. AB - In a dairy herd experiencing an abortion outbreak, 49% of the 119 cows were positive for Neospora caninum antibodies and the remaining 51% were seronegative. Using immunohistochemical analysis of foetal brain tissue, Neospora cysts and bradyzoite clusters were identified in two of the 15 aborted foetuses submitted for testing. Other agents that commonly cause abortions were not found. These results, coupled with neurological clinical symptoms in new-born calves, implicated N. caninum as the major cause of abortions in this dairy herd. This is the first report that associates N. caninum infection with bovine abortions in Portugal. PMID- 11928928 TI - Reproductive parameters and efficiency of inseminators in dairy farms in Portugal. AB - This study was carried out to determine the reproductive efficiency indices of one of the largest dairy co-operatives of northern Portugal, using data from 1980 to 1998. Records were made available by the computerized National Recording System. Age at first calving was 32.0 +/- 6.0 months. Mean calving to first AI interval was 95.4 +/- 30.0 days, and calving to conception intervals decreased (p < 0.05) from 176.9 +/- 4.5 to 148.1 +/- 5.6 days from the first to the fourth/fifth parturitions, respectively. Calving intervals decreased (p < 0.05) from 418.1 +/- 3.4 to 392.5 +/- 7.0 days from the first to fourth/fifth parturitions, respectively. Mean non-return rates at 90 days for first inseminations was 71.7 +/- 6.5% and mean calving rates at first insemination was 51.4 +/- 8.1%. There were significant differences (p < 0.001) in the inseminators' efficiency, measured by both non-return and calving rates at first AI, with differences between the best and worst results of 13.3 and 16.1% for non return and calving rates, respectively. The ranking of the inseminators did not coincide when their efficiency was measured by either non-return or calving rates. The mean number of inseminations per pregnancy (pregnant cows only) was 1.4 +/- 0.7 with significant (p < 0.001) differences among herds. The mean heat detection rate was 38.1 +/- 16.9%, with highly significant (p < 0.001) differences among farms (ranging from 14.2 to 60.8%). Negative (p < 0.001) correlations were found between heat detection rate and calving to first AI, calving to conception and calving intervals. The meaning of these indexes for assessment of reproductive efficiency in the studied system, is discussed. PMID- 11928929 TI - Relationship between motility and membrane integrity of boar spermatozoa in media varying in osmolality. AB - A study was conducted to evaluate the relationship between boar sperm motility and membrane integrity following exposure to media with 150-1120 mOsm. Total sperm motility was defined as the percentage of spermatozoa that had any form of motility was subjectively assessed under a light microscope. Sperm cell damage was expressed as a loss of membrane integrity as measured by a combination of fluorescent stains, carboxyfluorescein diacetate (CFDA) and propidium iodide (PI), and Hoechst 33258 (H33258). There were no significant differences between sperm motility and membrane-intact spermatozoa. as measured by CFDA-PI and H33258, in media with 250 and 300 mOsm. In anisosmotic conditions, a higher amount of membrane-intact spermatozoa than motile spermatozoa was observed. In hypo-osmotic conditions (150 mOsm), a high proportion of spermatozoa had curled or coiled tails and most of them retained their entire membrane integrity, as detected by CFDA-PI. In media with 350-1120 mOsm, some spermatozoa accumulated PI in the head region and CFDA in the mid-piece. These spermatozoa fluoresced blue at the lower region of the head, as detected by H33258. The ATP content in spermatozoa exposed to hypo- and hyperosmotic conditions was markedly reduced. There was no recovery of sperm motility on returning the spermatozoa to isosmotic conditions after 10 min incubation in anisosmotic conditions, indicating that the spermatozoa suffered an almost complete and irreversible loss of motility. This irreversible loss of motility may be a consequence of reduced ATP production in spermatozoa subjected to anisosmotic conditions. The results of this study demonstrate that plasma membrane integrity assessment in combination with sperm motility, using a range of media varying in osmolality, can give valuable information about the status and function of different sperm membranes, which might be relevant for semen preservation. PMID- 11928931 TI - Visible and valued. PMID- 11928930 TI - Defining nurses' ethical practices in the 21st century. PMID- 11928932 TI - Assessing patients' needs: does the same information guide expert and novice nurses? AB - The difficulties experienced by nurses in assessing patients before providing care have been the subject of enquiry for many years. Much has been written about the nursing process and, in particular, the data-collection component, where the nurse gathers information before deciding on a diagnosis and nursing intervention. There is, however, very little published on the differences between expert and novice nurses, in either the way they gather information or the emphasis placed on the different data sources accessed when preparing to carry out a nursing procedure. Communication between nurses is essential in the provision of safe, competent care, and yet we have minimal understanding of how experts use data sources to plan procedural care giving. This article reports on the findings of one component (the differences between expert and novice nurses in accessing data before implementing a nursing procedure) of a larger study into the identification of problem-solving strategies adopted by nurses during procedural care giving. The study was conducted in clinical settings and used a qualitative research methodology of observation followed by an in-depth semistructured interview. The study results indicate that expert and novice nurses accessed four similar information sources before meeting a patient. However, there were differences noted between the two groups in the amount of information accessed, as well as in the interpretation and use of that information. This is an important issue for nurse educators. PMID- 11928933 TI - Linking research and practice: participation of nurses in research to influence policy. AB - In this article, the authors describe research conducted by the Pakistan Nursing Council (PNC) in Islamabad, Pakistan. The research was carried out through collaboration of two components of a large multicomponent Canadian International Development Agency (CIDA)-funded project--the Development of Women Health Professionals Program (DWHP)--with the Colleges of Nursing and the Pakistan Nursing Council. The research was guided by staff of the DWHP and performed by eight nurses undergoing a research course as part of a Diploma in Teaching Administration (DTA) at a postgraduate College of Nursing in the Northwest Frontier Province (NWFP), Pakistan. Research questions related to the collection and analysis of nursing workforce statistics were asked and partially answered, while students gained experience in conducting research. A description of the context in which the research was conducted is provided. Finally, results of the research and the potential benefits for influencing health workforce policy are discussed. PMID- 11928934 TI - Quality of maternity care for adolescent mothers in Mbabane, Swaziland. AB - Early pregnancy and unplanned childbirth may have far-reaching physical, psychological and social consequences for the adolescent girl and her offspring and are therefore public health issues of concern. A number of evidence-based maternity practices might, if properly applied, prevent unnecessary health related problems in mothers and newborns, postnatally. In order to identify the areas of maternity practice that require improvement in Swaziland, the overall aim of this study was to generate systematic data on the maternity care and social support provided by health professionals (for adolescent mothers and their children) on admission, in the labour ward, and during and after delivery. The study was carried out during a 3-month period from April to June 1998. All pregnant adolescents with an uneventful term pregnancy, admitted to the Mbabane Government Hospital maternity ward in the morning of the study days, were informed about the purpose of the study and asked if they would like to participate. A total of 33 pregnant adolescents agreed and in-depth interviews were conducted with those participants. Observations and checklists were used to assess the maternity care given to the study participants. Results revealed that on admission to the labour ward, verbal communication and interaction between the midwife and the adolescent were minimal, and none of the adolescents was encouraged to bring a social support person to remain with them during labour. During the progress of labour, nearly 50% of the adolescent mothers developed complications and approximately 27% had a lower-segment Caesarean section. Special attention should therefore be paid to adolescent sexual and reproductive health service needs. These should include contraceptive counselling in order to prevent pregnancy at a young age and also to improve their sexual and reproductive health statuses. PMID- 11928935 TI - Reducing hatred and violence: is it our responsibility? PMID- 11928936 TI - From nurse leader to national legislator. A conversation with Asta Moller. PMID- 11928938 TI - ICN highlights family theme for International Nurses Day in May 2002. PMID- 11928937 TI - Individualized nursing care: an empirically generated definition. AB - Individualization is considered a particularly important feature of nursing care by nurses, patients and their families, and by health care administrators. Descriptions in the literature suggest that individualized care requires some background knowledge of the patient, which nurses use to devise care plans that treat each patient as unique. However, few research-based descriptions of individualized care exist. The purpose of this article was to offer a definition of individualized nursing care synthesized from research findings. Descriptions of nursing care individualization emerged from two previously conducted grounded theory studies of nursing practice. Data relevant to the concept were retrieved from each study. These data were compared and contrasted in order to identify and describe individualized care from the perspectives of nurses and patients. The analysis resulted in a definition of individualized nursing care that makes explicit the characteristics of such care, thus providing a framework for designing and evaluating nursing care that recognizes patients as unique persons. PMID- 11928939 TI - Stimulatory effects of hyperprolactinemia on aldosterone secretion in ovariectomized rats. AB - BACKGROUND: To evaluate the effects of hyperprolactinemia on aldosterone secretion and its mechanisms of action in ovariectomized (OVX) rats. METHODS: Hyperprolactinemia was induced by the transplantation of rat anterior pituitary (AP) glands under the kidney capsule for 6 weeks in female rats. Control rats underwent cerebral cortex (CX) transplantation. Four weeks after transplantation, the rats were OVX 2 weeks before decapitation. After decapitation, the trunk blood was collected, and the adrenal glands of CX- and AP-grafted rats were prepared as zona glomerulosa (ZG) cells for in vitro study. RESULTS: Plasma prolactin and aldosterone in the rats were increased by AP gland transplantation. In the in vitro study, the basal aldosterone secretion by the adrenal ZG cells was higher in AP-grafted rats than in CX-grafted rats. The AP-grafted group showed increased responsiveness to angiotensin II (10(-8) M), KCl (8 x 10(-3) M), or 8-bromo-adenosine 3',5'-cyclic monophosphate (8-br-cAMP; 10(-4) M, a membrane permeable analogue of cAMP) with regard to aldosterone secretion as compared with the CX-grafted group. N-(2-[p-Bromocinnamylamine]ethyl)-5-isoquinolinesulfonamide (H89; 10(-6), 10(-5) M, a protein kinase A inhibitor) or tetrandrine (10(-5) M, a blocker for both L-type and T-type Ca2+ channels) induced a greater suppression of aldosterone secretion in the AP-grafted group than in the CX-grafted group. No significant differences between the CX- and AP-grafted groups were observed, however, with regard to the adrenocorticotropichormone (10(-9) M)-, forskolin (10(-5) M, an adenylyl cyclase activator)-, or nifedipine (10(-5) M, an L-type Ca2+ channel blocker)-induced responsiveness of aldosterone secretion. In addition, there was no difference in the expression of desmolase (i.e., cytochrome P450 side-chain cleavage enzyme) in ZG cells between AP- and CX grafted rats. The conversions of 25-OH-cholesterol into pregnenolone in the presence of trilostane (an inhibitor of 3beta-hydroxysteroid dehydrogenase) and corticosterone into aldosterone, as well as the expression of the steroidogenic acute regulatory protein in ZG cells, were greater in AP-grafted rats than in CX grafted rats. CONCLUSIONS: These results suggest that hyperprolactinemia increases basal, angiotensin II- and KCl-stimulated aldosterone secretion by ZG cells in OVX rats through activation of T-type Ca2+ channels, the post-cAMP and protein kinase A pathway, cytochrome P450 side-chain cleavage enzyme, and aldosterone synthase, as well as by causing increased expression of steroidogenic acute regulatory protein in ZG cells. PMID- 11928940 TI - Acute effects of nitric oxide synthase inhibition on systemic, hepatic, and renal hemodynamics in patients with cirrhosis and ascites. AB - BACKGROUND: Nitric oxide synthase (NOS) inhibition has been demonstrated to correct systemic vasodilation and renal hypoperfusion in studies of patients with cirrhosis. In patients with decompensated cirrhosis, NOS blockade increases arterial pressure, but the acute effects on hepatic and renal hemodynamics are not known. METHODS: We examined the acute systemic, hepatic, and renal hemodynamic effects of N(G)-monomethyl-L-arginine (L-NMMA) in 10 patients with decompensated cirrhosis. After baseline measurements, 3 mg/kg L-NMMA was administered as an IV bolus. At 20 minutes, if mean arterial pressure did not increase by at least 10 mm Hg above the baseline value, a second injection of 6 mg/kg was administered. RESULTS: In 5 of 10 patients, the second injection of L NMMA 6mg/kg was necessary to achieve at least a 10 mm Hg increase in mean arterial pressure. Acute NOS inhibition increased systemic vascular resistance and decreased cardiac output, without causing changes in the hepatic venous pressure gradient. Hepatic blood flow decreased, but the indocyanine green intrinsic clearance and extraction remained unchanged. Plasma renin activity (from 9.5 +/- 2.9 to 6.7 +/- 1.6 ng/ml/h) and urinary prostaglandin E2 (from 299 +/- 40 to 112 +/- 36 pg/ml) significantly decreased. No significant changes in glomerular filtration rate, renal plasma flow, and natriuresis occurred, however. CONCLUSIONS: Acute L-NMMA infusion in patients with decompensated cirrhosis reduced hepatic blood flow and decreased plasma renin activity and urinary prostaglandin E2, without causing significant changes in renal hemodynamics. PMID- 11928941 TI - Pathogenic mechanism, prophylaxis, and therapy of symptomatic acidosis induced by acetazolamide. AB - BACKGROUND: Acetazolamide, a noncompetitive carbonic anhydrase inhibitor, can produce symptomatic acidosis and bone marrow suppression by a mechanism that is still unknown. This presentation occurs in the elderly, patients with renal or liver failure, people with diabetes, and newborns. The objective of this study was to understand the pathogenic mechanism of these adverse effects and to propose a possible prophylaxis and therapy. METHODS: Four human clinical cases were studied, and one animal experiment was performed. Four preterm newborns with posthemorrhagic ventricular dilation developed severe metabolic acidosis after treatment with acetazolamide. The acidosis suddenly disappeared after a packed red blood cell transfusion. Metabolic studies were performed in one patient and in newborn guinea pigs treated with 200 mg/kg acetazolamide. RESULTS: Acetazolamide can produce severe lactic acidosis with an increased lactate-to pyruvate ratio, ketosis with a low beta-hydroxybutyrate-to-acetoacetate ratio, and a urinary organic acid profile typical of pyruvate carboxylase deficiency. The acquired enzymatic injury resulting from the inhibition of mitochondrial carbonic anhydrase V that provides bicarbonate to pyruvate carboxylase can produce tricarboxylic acid cycle damage. We demonstrate that the dramatic disappearance of metabolic acidosis and normalizing metabolism after blood transfusion were due to the citrate contained in the packed red blood cell bag. This hypothesis was confirmed by animal experimentation. We argue that the metabolic disorder and bone marrow suppression may be related. CONCLUSION: We demonstrate how acetazolamide can lead to symptomatic metabolic acidosis and probably to bone marrow suppression. We suggest citrate as a possible prophylaxis and treatment for these adverse reactions. PMID- 11928942 TI - Atypical presentation of cocaine-induced Type A aortic dissection: a diagnosis made by transesophageal echocardiography. PMID- 11928943 TI - The JIM interview: Kenneth I. Shine, MD. PMID- 11928944 TI - AAMC Task Force issues first Financial Conflict of Interest Guidelines, and GAO urges HHS to reexamine financial conflict of interest guidance and regulations for NIH-funded research. Association of American Medical Colleges. U.S. General Accounting Office. Department of Health and Human Services. PMID- 11928945 TI - National Press Club hosts forum on disease research funding. PMID- 11928946 TI - New Rat Resource and Research Center to help investigators understand human disease. PMID- 11928947 TI - Effects of fasting on corticosterone production by zona fasciculata-reticularis cells in ovariectomized rats. AB - BACKGROUND: To investigate the function and mechanism of fasting on the production of corticosterone in vitro by zona fasciculata-reticularis (ZFR) cells from ovariectomized (OVX) rats. METHODS: Female rats were OVX for 4 days before decapitation. Rats were fed or fasted for 1 day before experiment. ZFR cells from fed and fasted rats were incubated with adrenocorticotropic hormone (ACTH), forskolin, 8-bromo-3',5'-cyclic adenosine monophosphate, SQ22536, nifedipine, chelerythrine chloride, trilostane or steroidogenic precursors at 37 degrees C for either 60 or 30 minutes. Corticosterone, pregnenolone concentrations in spent media, and the intracellular adenosine 3',5'-cyclic monophosphate (cAMP) concentration were determined by radioimmunoassay. The effects of fasting in response to ACTH on the protein expressions of steroidogenic acute regulatory protein (StAR) or cytochrome P450 side-chain cleavage enzyme (P450scc) in ZFR cells were determined by Western blot analysis. RESULTS: The concentration of plasma corticosterone in fasted rats was significantly higher than that in fed rats (P<0.01). One-day fasting significantly increased the responsiveness of ZFR cells to ACTH, forskolin, and precursor-stimulated corticosterone productions and to forskolin-stimulated cAMP accumulation. The corticosterone production was reduced in fasted group when adenylyl cyclase was inhibited by SQ22536. The fasting-enhanced level of corticosterone production in ZFR cells was decreased by the administration of nifedipine but not altered by that of chelerythrine chloride. Fasting significantly increased trilostane-stimulated production of pregnenolone in ZFR cells. The activities of enzymes which converting cholesterol, pregnenolone, progesterone, and deoxycorticosterone to corticosterone and the expressions of StAR in ZFR cells were greater in fasted rats than in fed rats. CONCLUSIONS: This study demonstrated that fasting increased the release of corticosterone and the accumulation of cAMP by rat ZFR cells. The action mediated through enhancing the responsiveness to ACTH stimulation, cAMP cascades and the activity of L-type calcium channels. The activities of steroidogenic enzymes including P450scc, 3beta-hydroxysteroid dehydrogenase, 21-hydroxylase, and 11beta-hydroxylase were all enhanced by the fasting treatment. PMID- 11928948 TI - New National Emphysema Treatment Trial paradigm of Health Care Financing Administration-sponsored clinical research trials: advances and dilemmas. PMID- 11928949 TI - Synthesis and distribution of cytokeratins in healthy and ulcerated bovine claw epidermis. AB - Keratinization of the epidermal cells of the bovine claw generates the horn that gives the tissue its mechanical strength. Disruption of keratinization is likely to have a detrimental effect on the strength and integrity of the horn, and could lead to solar lesions and lameness. As part of a wider investigation of the cell biological causes of lameness in dairy animals, we have compared keratin synthesis and distribution in healthy bovine claw tissue with those in hooves with solar ulcers. Protein synthesis was measured by [35S]-labelled amino acid incorporation in claw tissue explant cultures. [35S]-labelled protein synthesis was higher in tissue from diseased claws than in healthy claws, and highest at the ulcer site. The identity of proteins synthesised in vitro did not differ between healthy and diseased tissue. DNA synthesis indicative of cell proliferation was also elevated in diseased tissue. Immunoblotting after one- or two-dimensional electrophoresis showed cytokeratins (CK) 4, 5/6, 10 and 14 to be amongst those expressed in healthy claw tissue. The relative abundance of these keratins was not altered in healthy regions of ulcerated hooves, nor at the ulcer site, but CK16, not usually found in healthy tissue, was detected in the sole of diseased claws. CK5/6 and CK14 were shown by immunohistochemistry to be present in the basal epidermis of healthy tissue, whereas CK10 was found in supra-basal layers. In healthy tissue from ulcerated claws, this distribution was unaltered, but at the site of solar ulcers, CK5/6 and CK14 were each found in both basal and supra-basal epidermis. The study suggests that solar ulceration of the bovine claw is not associated with gross alteration in the keratin composition of the tissue, but causes abnormal distribution of cytokeratins, perhaps as a result of loss of positional cues from the basement membrane. Ulceration did, however, stimulate cell repair involving epidermal protein synthesis (including keratins), and keratinocyte proliferation. PMID- 11928950 TI - Decreased number and bactericidal activity against Staphylococcus aureus of the resident cells in milk of dairy cows during early lactation. AB - Phagocytic and bactericidal activity of polymorphonuclear neutrophil leukocytes (PMN) isolated from blood and milk, against Staphylococcus aureus, was compared between groups of six healthy dairy cows in early, mid- and late lactation using a bacteriological assay. PMN were isolated from blood with a high degree of purity, but the cells isolated from milk contained variable amounts of macrophages (Mphi) and lymphocytes (L). The results were therefore calculated using the percentage PMN in order to evaluate phagocytosis and killing by PMN only. Blood PMN phagocytosed 82% Staph. aureus and milk PMN 43% on average and there was no significant difference between the different stages of lactation. The bactericidal activity of blood PMN against Staph. aureus was 36+/-8% in early lactation (significantly different from mid lactation, P < 0.05), 64+/-10% in mid lactation and 53+/-6% in late lactation. Milk PMN killed only 6+/-3% Staph. aureus in early lactation (significantly different from mid lactation, P < 0.01), 27+/-3% in mid lactation and 20+/-9% Staph. aureus in late lactation. The ratio of the bactericidal activity of milk to blood PMN was 0.08, 0.43 and 0.22 in early, mid- and late lactation, respectively. In addition to the decreased function. the number of cells in milk (somatic cell count, SCC) was also 60% lower in early lactation than in mid lactation cows (P < 0.01). Our results suggest an impairment of blood and milk-resident PMN bactericidal activity against Staph. aureus and a decreased number of milk-resident PMN in dairy cows at the onset of lactation. PMID- 11928951 TI - Effect of localised antibiotic infusions applied to the teat-canal and teat sinus at drying-off on mastitis in the dry-period and at calving. AB - An experiment using three New Zealand herds and a total of 632 cows, examined the effect of localised prophylactic treatments with antibiotic at drying-off on the incidence of new intramammary infection during the dry period and at calving. Antibiotic was infused either into the teat canal (0.22 g of dry-cow formulation) or the teat sinus (3.1 g of lactating-cow formulation) of uninfected quarters to eliminate any bacteria present in these locations at the last milking of lactation. These treatments were compared with a negative control (nil treatment) and a positive antibiotic control (infusion of 3.6 g of dry-cow formulation). All antibiotic formulations used the same active ingredient, sodium cloxacillin. No significant reduction in new dry period clinical mastitis was observed for the two localised treatments whereas the positive control treatment achieved 100% reduction in new clinical mastitis compared with untreated control quarters. A 41% reduction (P < 0.05) in new Streptococcus uberis infections at calving was associated with the teat canal antibiotic treatment, compared with an 82% reduction (P < 0.001) for the positive antibiotic control. Both localised treatments showed a reduced incidence of new intramammary infection (P < 0.001) when pooled across periods and pathogens. Teats receiving either the teat canal antibiotic treatment or a full infusion of long acting dry-cow antibiotic had a lower incidence of open teat canals (P < 0.05) at 3 weeks after drying-off. PMID- 11928952 TI - Inhibition of oxytocin release and milk let-down in postpartum primiparous cows is not abolished by naloxone. AB - About 10% of primiparous cows have no milk ejection during the first milkings after delivery. Therefore, 17 Brown Swiss dairy cows in their first lactation were used to evaluate the extent of disturbed milk let-down and the corresponding oxytocin (OT) plasma values in the 1st 5 days after delivery. The first milking was 9-22 h after parturition and served for classification of the cows to groups with inhibited (INH), bimodal (BIMO), or normal (NOR) milk let-down. The OT plasma levels before the start of manual teat stimulation and machine milking were comparably high during the first milking especially in NOR and BIMO cows. Ten minutes before the second milking (M2), 300 mg of the opioid antagonist naloxone was injected to test whether the disturbance was affected by the action of endogenous opioids on the neurohypophysis. The milk yield was not influenced by the naloxone treatment, and the INH cows had milk ejection only after a vaginal stimulation. Afterwards, the cows were milked twice every day, until the milk let-down and the OT release were unaffected (equal to control milking). Then, at the next milking, the cows were injected with 300 mg morphine 10 min before milking. The central OT release in response to manual teat stimulation and machine milking was completely blocked in all cows, but a vaginal stimulation was able to abolish this block, at least partially, in 16 cows. Thus, morphine produced a milk let-down characteristic as in the INH cows during the first three milkings. For the following milking, the cows were pre-treated with 300 mg naloxone (-15 min) plus 300 mg morphine (-10 min) before milking. The OT release and the milk yields were unaffected when compared with the control milking. This experiment demonstrates that exogenous opioids can affect the central release of OT in a naloxone-reversible manner even very soon after parturition. However, endogenous opioids are probably not the main mediators of disturbed central OT release and alveolar milk ejection in post-partum primiparous cows. PMID- 11928953 TI - Effect of genetic potential and level of feeding on milk protein composition. AB - Two groups of 15 multiparous cows in mid-lactation were used in a Latin square design experiment with 4-week experimental periods. The genetic milk protein concentration level was high in the first group and low in the second. Each group of cows was given in a random order three feeding levels that covered 85, 100 and 115% of energy requirements and 90, 110 and 125% of nitrogen requirements, respeetively. In both groups, increasing level of feeding induced a significant increase in milk yield (+ 2.4 kg/d between lowest and highest levels) and in protein concentration (+ 1.7 g/kg). The proportion of paracasein in total proteins was not altered by either genetics or nutrition. The proportion of casein in total proteins was slightly increased by 0.5 percentage points (P < 0.05) with the intermediate level of feeding. Plasmin and plasminogen activities were not significantly modified by the genetic milk concentration level. Plasmin activity significantly increased with nutrient supplementation, but only in animals of low genetic potential (+ 21% between low and high levels, P < 0.01). Casein composition was not significantly altered by the genetics or level of nutrition. Over the whole range of individual measurements taken (n = 90), the relationships between casein or paracasein and total protein concentrations were linear and very narrow (R2 = 0.92 and 0.95, respectively). The proportion of casein or paracasein in total proteins significantly decreased as plasmin activity increased. PMID- 11928954 TI - Effect of beta-lactoglobulin on plasma retinol and triglyceride concentrations, and fatty acid composition in calves. AB - Beta-lactoglobulin (beta-lg) is the main protein of ruminant milk whey. Although beta-lg can bind in vitro to a variety of hydrophobic substrates, mainly retinol and long-chain fatty acids, its physiological function is still unknown. In Exp. 1, we investigated the effect of beta-lg on the plasma retinol concentration in preruminant calves. Holstein male calves (n = 20) were fed Holstein whole milk at 40 g/kg body weight (BW) plus vitamin A acetate (500,000 i.u.) with or without beta-lg (0.4 g/kg BW). The plasma retinol concentration of 10-d-old calves was greater (P < 0.05) in the beta-lg-fed group) than in the control group during the period from 8 to 12 h and at 24 h after the feeding. The postprandial change of plasma retinol in 40-d-old calves fed milk with beta-lg was higher (P < 0.05) than that in the control calves only at 12 h after the feeding. In Exp. 2, Holstein male calves (n = 18) were used to investigate the effect of beta-lg on plasma triglyceride concentration and fatty acid composition. Calves were fed Holstein whole milk at 40 g/kg BW plus milk fat prepared from whole milk at 2 g/kg BW with or without beta-lg (0.4 g/kg BW). Plasma triglyceride concentration at age 10 d was higher (P < 0.05) in the beta-lg-fed group than in the controls during the periods from 1 to 2 h and from 7 to 11 h after the feeding. At age 40 d, plasma triglyceride in the beta-lg-fed group was higher (P < 0.05) than in the control group only at 9 h. Ratios of palmitic, stearic, and oleic acids to total plasma lipids were higher (P < 0.05) in the calves fed beta-lg milk than in the control calves at age 10 d. These results suggest that beta-lg enhances the intestinal uptake of retinol, triglyceride, and long-chain fatty acids in preruminant calves. PMID- 11928955 TI - Immune modulation in suckling rat pups by a growth factor extract derived from milk whey. AB - Oral tolerance to foreign enteral antigens is not fully developed in early neonatal life. Epidemiological evidence supports a role for maternal milk in the development of immune responses, including oral tolerance. Formula fed infants have an increased susceptibility to food allergy and the later development of autoimmune disease. This may relate to the lack in infant formula of growth factors found in maternal milk. Bovine milk contains proteins, growth factors and cytokines. Various studies have outlined the immune modulating potential of bovine milk-derived products. Fractionated whey extracts have therapeutic potential in disease states where there is an excessive inflammatory reaction, and disease preventive potential for infants who are not breast-fed. We have shown that daily oral administration of a growth factor-enriched fraction from milk whey to naturally suckling rat pups between days 4-9 postnatal can down regulate immune activation to a specific orally administered food antigen, ovalbumin, assessed by lymphocyte proliferation. In addition, non-specific down regulation in the intestine was observed as assessed by the expression of MHC I. Treatment of rat pups with whey extract at the time of oral sensitisation to ovalbumin also resulted in an increased secretion of TGF-beta into the culture supernatant of spleen cells incubated with specific antigen. TGF-beta is an immuno-down-regulatory cytokine involved in tolerance induction. Immune modulation by extracts derived from milk whey could be of potential benefit for formula-fed and pre-term infants in reducing susceptibility to inappropriate activation to food antigens. PMID- 11928956 TI - Immunomodulating effects of milks fermented by Lactobacillus helveticus and its non-proteolytic variant. AB - The effect of milks fermented by Lactobacillus helveticus and its non-proteolytic variant on mucosal and tumoral immunity was studied. Milks fermented by Lb. helveticus wild type or its non-proteolytic variant were administered orally to mice for different periods (3, 5 and 7 d). The immune response was assessed by analysing the activity of the peritoneal macrophages, the number of cells secreting IgA associated with the gut-associated lymphoid tissue and with the bronchial-associated lymphoid tissue. The number of cells was determined by direct immunofluorescence. The antitumour activity was monitored by studying the regression of the subcutaneously implanted fibrosarcomas. After 3 d feeding of milk fermented by Lb. helveticus wild type, the number of sIgA increased significantly at both the intestinal and bronchial levels, indicating that a cellular migration had occurred. This effect was not noticeable when milk fermented by Lb. helveticus Protease (-) was orally administered. Both fermented milks (wild type or its variant) exhibited an effect on the activity of the peritoneal macrophages, which might be indirectly correlated to the regression of the fibrosarcoma. Although the mechanism by which the lactic acid bacteria enhance the immune system is not clear, this study clearly suggests that the bioactive compounds released during milk fermentation might contribute to the immunoenhancing properties of these products. By releasing biopeptide, lactic acid bacteria have important implications in modulation of the host's immune response, more specifically its cellular immune response. PMID- 11928957 TI - Immune enhancement conferred by oral delivery of Lactobacillus rhamnosus HN001 in different milk-based substrates. AB - The probiotic Lactobacillus rhamnosus HN001 is known to enhance immunity in animal models, following oral delivery. In this study, we investigated the immunoenhancing effects of HN001 delivered to mice in different milk-based substrates, including: whole (full-fat) milk supplemented with HN001; fermented milk supplemented with HN001; or whole milk which had been part-fermented by HN001. Direct oral feeding of mice with HN001 in whole milk was shown to enhance the phagocytic activity of blood and peritoneal cells. Similar effects on phagocytosis were observed when UN001 was offered to mice in the form of a milk- or fermented milk-based diet. The degree of immune enhancement conferred by HN001 was similar whether the probiotic was used as an additive or as a fermentative agent. These studies confirm that Lb. rhamnosus HN001, derived originally from dairy food, enhances immune function following oral delivery in different milk bases. PMID- 11928958 TI - Hypocholesterolemic effect of Lactobacillus gasseri SBT0270 in rats fed a cholesterol-enriched diet. AB - The effects of administration of Lactobacillus gasseri SBT0270 on serum lipids and bile acids, faecal bile acids and microflora were estimated in hypercholesterolemic rats. An effective dose of strain SBT0270 to exert its hypocholesterolemic effect was 10(9) viable cells/d. The dose of 10(9) cells/d did not affect the faecal coliform counts, but the number of faecal lactobacilli in rats fed this dose was significantly higher than that in the control group observed at the end of feeding period. Hypocholesterolemic effect of Lb. gasseri SBT0270 was attributed to its ability to suppress the reabsorption of bile acids into the enterohepatic circulation and to enhance the excretion of acidic steroids in faeces of hypercholesterolemic rats. PMID- 11928959 TI - Effect of temperature and/or pressure on lactoperoxidase activity in bovine milk and acid whey. AB - At atmospheric pressure, inactivation of lactoperoxidase (LPO) in milk and whey was studied in a temperature range of 69-73 degrees C and followed first order kinetics. Temperature dependence of the first order inactivation rate constants could be accurately described by the Arrhenius equation, with an activation energy of 635.3 +/- 70.7 kJ/mol for raw bovine milk and 736.9 +/- 40.9 kJ/mol for diluted whey, indicating a very high temperature sensitivity. On the other hand, LPO is very pressure resistant and not or only slightly affected by treatment at pressure up to 700 MPa combined with temperatures between 20 and 65 degrees C. Both for thermal and pressure treatment, stability of LPO was higher in milk than in diluted whey. Besides, a very pronounced antagonistic effect between high temperature and pressure was observed, i.e. at 73 degrees C, a temperature where thermal inactivation at atmospheric pressure occurs rapidly, application of pressure up to 700 MPa exerted a protective effect. At atmospheric pressure, LPO in diluted whey was optimally active at a temperature of about 50 degrees C. At all temperatures studied (20-60 degrees C), LPO remained active during pressure treatment up to 300 MPa, although the activity was significantly reduced at pressures higher than 100 MPa. The optimal temperature was found to shift to lower values (30-40 degrees C) with increasing pressure. PMID- 11928960 TI - Chemical and microbiological characterisation of kefir grains. AB - Chemical and microbiological composition of four Argentinean kefir grains from different sources as well as characteristics of the corresponding fermented milk were studied. Kefir grains CIDCA AGK1, AGK2 and AGK4 did not show significant differences in their chemical and microbiological composition. In contrast, protein and yeast content of AGK3 was higher than in the other grains. Although grain microflora comprised lactobacilli, lactococcus, acetic acid bacteria and yeast, we found an important difference regarding species. Lactococcus lactis subsp. lactis, Lactobacillus kefir, Lactobacillus plantarum, Acetobacter and Saccharomyces were present in all types of kefir grain. While Leuconostoc mesenteroides was only isolated from grains CIDCA AGK1 and Lactococcus lactis subsp. lactis biovar diacetylactis, Lactobacillus parakefir and Kluyveromyces marxianus were only isolated from CIDCA AGK2 grains. All grains produced acid products with pH between 3.5 and 4.0. The apparent viscosity of AGK1 fermented milk was greater than the product obtained with AGK4. All fermented milks had inhibitory power towards Escherichia coli but AGK1 and AGK2 supernatants were able to halt the bacterial growth for at least 25 h. Grain weight increment in AGK1, AGK2 and AGK3 during growth in milk did not show significant differences. Despite their fermenting activity, AGK4 grains did not increase their weight. PMID- 11928961 TI - Polysaccharide production by kefir grains during whey fermentation. AB - Fermentation of deproteinised whey with kefir grains CIDCA AGK1 was studied focusing on polysaccharide production from lactose. Kefir grains were able to acidify whey at different rates depending on the grain/whey ratio. During fermentation, kefir grains increased their weight and a water-soluble polysaccharide was released to the media. Exopolysaccharide concentration increased with fermentation time, reaching values of 57.2 and 103.4 mg/l after 5 days of fermentation in cultures with 10 and 100 g kefir grains/l, respectively. The polysaccharide fraction quantified after fermentation corresponded to the soluble fraction, because part of the polysaccharide became a component of the grain. Weight of kefir grains varied depending on the time of fermentation. Polysaccharide production was affected by temperature. Although the highest concentration of polysaccharide in the media was observed at 43 degrees C at both grain/whey ratios, the weight of the grains decreased in these conditions. In conclusion, kefir grains were able to acidify deproteinised whey, reducing lactose concentration, increasing their weight and producing a soluble polysaccharide. PMID- 11928962 TI - L-methionine degradation potentialities of cheese-ripening microorganisms. AB - Volatile sulphur compounds are major flavouring compounds in many traditional fermented foods including cheeses. These compounds are products of the catabolism of L-methionine by cheese-ripening microorganisms. The diversity of L-methionine degradation by such microorganisms, however, remains to be characterized. The objective of this work was to compare the capacities to produce volatile sulphur compounds by five yeasts, Geotrichum candidum, Yarrowia lipolytica, Kluyveromyces lactis, Debaryomyces hansenii, Saccharomyces cerevisiae and five bacteria, Brevibacterium linens, Corynebacterium glutamicum, Arthrobacter sp., Micrococcus lutens and Staphylococcus equorum of technological interest for cheese-ripening. The ability of whole cells of these microorganisms to generate volatile sulphur compounds from L-methionine was compared. The microorganisms produced a wide spectrum of sulphur compounds including methanethiol, dimethylsulfide, dimethyldisulfide, dimethyltrisulfide and also S-methylthioesters, which varied in amount and type according to strain. Most of the yeasts produced methanethiol, dimethylsulfide, dimethyldisulfide and dimethyltrisulfide but did not produce S methylthioesters, apart from G. candidum that produced S-methyl thioacetate. Bacteria, especially Arth. sp. and Brevi. linens, produced the highest amounts and the greatest variety of volatile sulphur compounds includling methanethiol, sulfides and S-methylthioesters, e.g. S-methyl thioacetate, S-methyl thiobutyrate, S-methyl thiopropionate and S-methyl thioisovalerate. Cell-free extracts of all the yeasts and bacteria were examined for the activity of enzymes possibly involved in L-methionine catabolism, i.e. L-methionine demethiolase, L methionine aminotransferase and L-methionine deaminase. They all possessed L methionine demethiolase activity, while some (K. lactis, Deb. hansenii, Arth. sp., Staph. equorum) were deficient in L-methionine aminotransferase, and none produced L-methionine deaminase. The catabolism of L-methionine in these microorganisms is discussed. PMID- 11928963 TI - Determination of taste-active compounds of a bitter Camembert cheese by omission tests. AB - The taste-active compounds of a Camembert cheese selected for its intense bitterness defect were investigated. The water-soluble fraction (WSE) was extracted with pure water and fractionated by successive tangential ultrafiltrations and nanofiltration. The physicochemical assessment of these fractions led to the construction of a model WSE which was compared by sensory evaluation to the crude water-soluble extract, using a panel of 16 trained tasters. As no significant difference was perceived, this model WSE was then used directly or mixed with other cheese components for omission tests. Among the main taste characteristics of the WSE (salty, sour, umami and bitter), bitterness was found to be due to small peptides whose mass distribution was obtained by RPHPLC MS (400-3000 Da) and whose taste properties are discussed. PMID- 11928965 TI - Alpha1c- and beta2-adrenergic receptor mRNA distribution in the bovine mammary gland detected by competitive RT-PCR. PMID- 11928964 TI - Use of duplex polymerase chain reaction (duplex-PCR) technique to identify bovine and water buffalo milk used in making mozzarella cheese. AB - Molecular biology techniques have been used for species identification in food of animal origin in relatively recent years. A polymerase chain reaction (PCR) based method, the multiplex PCR, was recently applied to species identification in meat and meat products. It allows co-amplification of separate regions of a single gene or specific fragments, each typical of a different animal species in a single PCR reaction, using different pairs of primers in the same reaction mix. In the present paper, the duplex-PCR technique is proposed to identify bovine and water buffalo DNA in a single PCR assay in milk and mozzarella cheese (a typical Italian cheese, originally made from pure water buffalo milk). Because of its lower cost, undeclared bovine milk is added to water buffalo milk for making different kinds of mozzarella cheese. The results of this experiment indicate the applicability of this method, which showed an absolute specificity for the two species and a high sensitivity even down to low DNA concentrations (1 pg). In bovine and water buffalo mixtures of both milk and mozzarella cheese, the minimum concentration tested was 1% of bovine in water buffalo milk and water buffalo in bovine milk. The importance of the somatic cell content in raw milk is also discussed with special reference to the evaluation of mixtures (milk or cheese) of the two species. PMID- 11928966 TI - Evaluation and other issues. PMID- 11928967 TI - Controlled evaluation of a community based injury prevention program in Australia. AB - OBJECTIVE: To evaluate the effects of a community based, all age, all injury prevention program, the Safe Living Program, on injury risk and injury rates. DESIGN: A quasiexperimental population based evaluation using an intervention and comparison community design. SETTING: The intervention community (Shire of Bulla, n = 37,257) is an outer metropolitan area of Melbourne, Australia. The demographically matched comparison community (Shire of Melton, n=33,592) is located nearby. SUBJECTS AND METHODS: The Safe Living Program in the Shire of Bulla targeted injury reduction in all settings with a focus on high risk groups. Strategies included program publicity, education and training, injury hazard reduction, and environmental change. Baseline and follow up measures of program reach, risk factors, and injury rates in both communities were used to evaluate program process, impact, and outcome. RESULTS: Increase in program awareness was moderate and similar to other community based programs. The program achieved injury hazard reduction on the road, in schools, and, to a more limited extent, in the home. Other changes in injury risk factors could not necessarily be attributed to the program as similar changes were observed in the comparison community. No significant changes were found in rates of injury deaths, hospitalisations, or emergency department presentations in the Shire of Bulla after six years. Self reported household injuries, mostly minor, were reduced in the intervention community, but had been higher at program launch than in the comparison community. CONCLUSIONS: The Safe Living Program was unable to replicate the significant reductions in injuries reported in other community based interventions. Replication of apparently successful community based injury prevention programs in different settings and populations requires evidence based interventions, sustained and effective program penetration, reliable data systems to measure change, at least one control community, and sufficient budget and time for effects to be observable. PMID- 11928968 TI - Evaluation of a child safety program based on the WHO safe community model. AB - OBJECTIVES: To evaluate the outcome of the World Health Organization (WHO) Safe Community model with respect to child injuries. STUDY DESIGN: A population based quasiexperimental design was used. Cross sectional pre-implementation and post implementation data were collected in intervention (Motala municipality) and control (Mjolby municipality) areas, both in Ostergotland county, Sweden. RESULTS: The total relative risk of child injury in the intervention community decreased more (odds ratio 0.74; 95% confidence interval (CI) 0.68 to 0.81) than in a control community exposed only to national level injury prevention programs (0.93; 95% CI 0.82 to 1.05). The relative risk of moderately (abbreviated injury scale (AIS) 2) severe injury in the study area was reduced to almost a half (odds ratio 0.49; 95% Cl 0.41 to 0.57), whereas the risk of minor (AIS 1) injuries decreased only slightly (odds ratio 0.89; 95% CI 0.80 to 0.99). The risk of severe or fatal (AIS 3-6) injuries remained constant. CONCLUSIONS: After introduction of an injury prevention program based on the WHO Safe Community model, the relative risk for child injury in the intervention community decreased significantly more than in a control community exposed only to national injury prevention programs. PMID- 11928969 TI - Influence of safety gear on parental perceptions of injury risk and tolerance or children's risk taking. AB - OBJECTIVES: Risk compensation theory has been shown to relate to how individuals behave in areas such as traffic safety and consumer product safety. The present study examines whether risk compensation theory applies to parents' judgments about school age children's permissible risk taking under non-safety gear and safety gear conditions for seven common play situations. The extent of the child's experience with the activity and parental beliefs about safety gear efficacy were examined as possible moderators of extent of children's risk taking allowed by parents. METHOD: A telephone interview was used to obtain each parent's ratings of permissible risk taking by their child (for example, speed at which child is allowed to cycle, height allowed to climb to on a climber) under safety gear and no gear conditions, and ratings of child experience and gear efficacy. RESULTS: Results confirmed risk compensation operated under all seven play situations, resulting in parents reporting they would allow significantly greater risk taking by their children under safety gear than non-safety gear conditions. Children with more experience with the activities were to be allowed greater risk taking, even when not wearing safety gear. Parents who believed more strongly in the efficacy of the safety gear to prevent injuries showed greater risk compensation. No sex differences emerged in any analyses. CONCLUSION: Results highlight the need to communicate to parents that safety gear moderates injury risk but does not necessarily guarantee the prevention of injury, particularly if children are allowed greater risk taking when wearing safety gear. PMID- 11928970 TI - Influence of free trade on the politics of safety. PMID- 11928971 TI - Adolescent injury morbidity in New Zealand, 1987-96. AB - OBJECTIVES: Adolescents are over-represented in injury statistics. New Zealand is privileged in having a hospital discharge database allowing for analysis of non fatal injury data at a national level. An epidemiological description of adolescent injury morbidity is provided and options for prevention are discussed. METHOD: People aged 15-19 years admitted to hospital for their injuries in the period 1 987-96 were identified from the New Zealand Health Information Service morbidity data files. The manner, causes, and nature of injury were examined. Injury prevention strategies were reviewed. RESULTS: The incidence of hospitalised injury was 1,886 per 100,000 person years. The victims were male (70%). The leading causes of injury were road traffic crashes, sports injuries, and self poisoning. The most common injury diagnoses were head injuries (29%) and limb fractures (21%). Road traffic crashes produced the highest proportion of serious injuries. CONCLUSIONS: Road traffic crashes, sports injuries, and self inflicted poisoning, stood out as areas with the greatest potential for reducing the burden of injury in late adolescence. Graduated driver licensing shows promise as an injury prevention measure but remains inadequately implemented. Policies to reduce self inflicted poisoning are of unknown efficacy, and evidence is awaited on the effectiveness of measures to reduce injury in sport. PMID- 11928972 TI - Risk factors for burns in children: crowding, poverty, and poor maternal education. AB - OBJECTIVE: To characterize the presentation of burns in children and risk factors associated with their occurrence in a developing country as a basis for future prevention programs. DESIGN: Case-control study. SETTING: Burn unit of the National Institute of Child Health (Instituto Nacional de Salud del Nino) in Lima, Peru. METHODS: A questionnaire was administered to all consenting guardians of children admitted to the burns (cases) and general medicine (controls) units during a period of 14 months. Guardians of patients were questioned regarding etiology of the injury, demographic and socioeconomic data. RESULTS: 740 cases and controls were enrolled. Altogether 77.5% of the cases burns occurred in the patient's home, with 67.8% in the kitchen; 74% were due to scalding. Most involved children younger than 5 years. Lack of water supply (odds ratio (OR) 5.2, 95% confidence interval (CI) 2.1 to 1 2.3), low income (OR 2.8, 95% CI 2.0 to 3.9), and crowding (OR 2.5, 95%CI 1.7 to 3.6) were associated with an increased risk. The presence of a living room (OR 0.6, 95% CI 0.4 to 0.8) and better maternal education (OR 0.6, 95% CI 0.5 to 0.9) were protective factors. CONCLUSIONS: To prevent burns interventions should be directed to low socioeconomic status groups; these interventions should be designed accordingly to local risk factors. PMID- 11928973 TI - Effects of state helmet laws on bicycle helmet use by children and adolescents. AB - OBJECTIVE: To evaluate the effectiveness of state helmet laws in increasing the use of bicycle helmets by children and adolescents under age 16. SETTING: United States. METHODS: A cross sectional study of factors associated with the likelihood of helmet use by children and adolescents. Data were derived from a national random digit dial telephone survey of bicycle riders. A multiple logistic regression analysis was used to quantify the independent effect of the state helmet laws on helmet use. RESULTS: Helmet use was systematically related to the presence of state helmet laws (odds ratio 2.65; 95% confidence interval (CI) 1.29 to 5.44). The increase in the average probability of helmet use attributable to state helmet laws was 18.4% (95% CI 17.8% to 19.0%). CONCLUSIONS: State helmet laws significantly increase helmet use by children and play an important part in any comprehensive effort designed to achieve this goal. PMID- 11928974 TI - State level estimates of the incidence and economic burden of head injuries stemming from non-universal use of bicycle helmets. AB - OBJECTIVE: To develop national and state level estimates for preventable bicycle related head injuries (BRHIs) and associated direct and indirect health costs from the failure to use bicycle helmets. METHODS: Information on the effectiveness and prevalence of use of bicycle helmets was combined to estimate the avoidable fraction, that is, the proportion of BRHIs that could be prevented through the use of bicycle helmets. The avoidable fraction multiplied by the expected number of BRHIs gives an estimate of the number of preventable cases. Direct and indirect health costs are estimated from a social perspective for the number of preventable BRHIs to assess potential cost savings that would be achieved if all riders wore helmets. RESULTS: Approximately 107,000 BRHIs could have been prevented in 1997 in the United States. These preventable injuries and deaths represent an estimated $81 million in direct and $2.3 billion in indirect health costs. Estimates range from 200 preventable BRHIs and $3 million in health costs in Wyoming (population 480,000) to 13,700 preventable BRHIs and $320 million in health costs in California (population 32.3 million). CONCLUSIONS: A number of successful approaches to increasing bicycle helmet use exist, including mandatory use laws and community based programs. The limited use of these strategies may be related to the fact that too little information is available to state agencies about the public health and economic burden of these preventable injuries. In conjunction with information on program costs, our estimates can assist state planners in better quantifying the number of preventable BRHIs and the costs and benefits of helmet promotion programs. PMID- 11928975 TI - Understanding the knowledge and attitudes of commercial drivers in Ghana regarding alcohol impaired driving. AB - OBJECTIVES: The knowledge and attitudes of commercial drivers in Ghana as regards alcohol impaired driving were investigated. This was done in order to provide information that could subsequently be used to develop antidrunk driving social marketing messages built upon the intrinsic values and motivation of these drivers. METHODS: Focus group discussions were held with 43 bus and minibus drivers in the capitol city, Accra. A structured discussion guide was used to capture information related to values, risk perceptions, leisure time activities, and attitudes on alcohol impaired driving. RESULTS: The majority of drivers expressed an understanding that drunk driving was a significant risk factor for crashes. There was a significant under-appreciation of the extent of the problem, however. Most believed that it was only rare, extremely intoxicated drivers who were the problem. The drivers also had a minimal understanding of the concept of blood alcohol concentration and related legal limits. Despite these factors, there was widespread support for increased enforcement of existing antidrunk driving laws. CONCLUSIONS: In Ghana, commercial drivers understand the basic danger of drunk driving and are motivated to assist in antidrunk driving measures. There are misconceptions and deficits in knowledge that need to be addressed in subsequent educational campaigns. PMID- 11928976 TI - Patterns of violence in Karachi, Pakistan. AB - OBJECTIVES: Government reports on violence in developing countries are almost universally based upon police data, which are typically incomplete and unreliable. Violence in Karachi was evaluated using ambulance service, not police, records. SETTING: Karachi, Pakistan. METHODOLOGY: The study was based on a case series of persons suffering from intentional injuries and transported by Edhi, the largest emergency service in Karachi, between October 1993 and January 1996. Main outcome variables were injury and death rates. RESULTS: Edhi Ambulance Service transported 4,091 intentionally injured persons during the 29 month period from October 1993 to January 1996. Ninety five per cent (n=3,864) were males; 74% (n=2,823) were 20 to 40 years of age, and 2,400 (58%) died before reaching the hospital. Firearms were the most common mode of injury (n=3,396, 83%). Forty six per cent (n= 1,828) of violent injuries occurred in four neighborhoods of Karachi--22% in Korangi (n= 884) and 8% each in Orangi (n=337), Malir Colony (n=307), and Nazimabad (n=300). On the 32 days when a political strike was called, more people were injured (mean = 10.4 v 5.0 persons, p=0.01) and killed (mean 6.6 v 3.9 persons, p<0.01) compared with days without a political strike. CONCLUSION: Violence is a major public health problem in Karachi, affecting predominantly wage earners. At least some of the violence is rooted within the political system. Detailed study of the causes of violence that explains the role that political and ethnic tensions play, may suggest strategies to lessen the toll of violence. PMID- 11928977 TI - Statistical and design issues in studies of groups. PMID- 11928978 TI - Recreational injury and its relation to socioeconomic status among school aged children in the US. AB - OBJECTIVES: This study described epidemiologic patterns of recreational injuries among school aged children in the US and assessed the relation of these patterns to socioeconomic status. METHODS: Combined data from the 1997-98 National Health Interview Surveys for 38 458 children aged 6-17 years regarding non-fatal recreational injury episodes that received medical attention, reported by a household adult, were analysed. Logistic regression analysis was used to assess the association between recreational injury and socioeconomic status while controlling for confounding factors. RESULTS: The annualized rate of recreational injury was 91.2 episodes per 1,000 children, with an increased risk associated with a higher family income status or being non-Hispanic white. For children from not poor families, most injury episodes occurred in sport facilities, whereas for children from poor and near poor families, most occurred outside the home. CONCLUSION: Recreational injury is a significant health problem for school aged children in the US. Non-Hispanic white children and children from affluent families are at increased risk of recreational injury. PMID- 11928979 TI - The New Zealand Blood Donors' Health Study: baseline findings of a large prospective cohort study of injury. AB - INTRODUCTION: Cohort studies have contributed important scientific knowledge regarding the determinants of chronic diseases. Despite the need for etiologic investigations, this design has been infrequently used in injury prevention research. OBJECTIVES: To describe the baseline findings of the New Zealand Blood Donors' Health Study, a large prospective study designed to investigate relationships between lifestyle, psychosocial factors, and serious injury due to road crashes, falls, self harm, assault, work, sport, and recreation. METHODS: Participants were recruited from fixed and mobile collection sites of a voluntary non-profit blood donor program. Baseline exposure data (for example risk taking behaviors, alcohol and marijuana use, sleep habits, and depression) were collected using a self administered questionnaire. Outcome data regarding serious injury will be collected prospectively through computerized record linkage of participants' unique identifiers to national morbidity and mortality databases. RESULTS: In total, 22 389 participants enrolled in the study (81% response rate). The diverse study population included 36% aged 16-24 years, 20% rural residents, and large variability in exposures of interest. For example, in the 12 months before recruitment, 21% had driven a motor vehicle when they considered themselves over the legal limit for alcohol, and 11% had been convicted of traffic violations (excluding parking infringements). Twelve per cent had seriously considered attempting suicide sometime in their life. CONCLUSIONS: This is the first, large scale cohort study investigating determinants of serious injury in New Zealand and among the largest worldwide. Preliminary findings from prospective analyses that can inform injury prevention policy are expected within five years. PMID- 11928980 TI - Changes in injury mortality by intent and mechanism in Taiwan, 1975-98. AB - BACKGROUND: Most official mortality publications do not present the mechanism of injury (for example, cut/pierce, drowning, fall, poisoning, or suffocation) for intentional injuries (for example, suicide or homicide). OBJECTIVES: To determine if the presentation of mechanism of injury for intentional injuries had different mechanism profiles. METHODS: Age adjusted injury mortality rates by intent and mechanism of injury for Taiwan were calculated for the years 1975 to 1998. The International Classification of Disease codes for the matrix by intent and mechanism groupings were based on recommendations of the US Centers for Disease Control and Prevention. RESULTS: If rates for both groups (intentional and unintentional) are combined, the importance of poisoning and suffocation increase relative to their contribution for unintentional injuries alone. Given the same mechanism of injury, different intents showed different patterns of change during the study period and given the same intent, the changes over time differed for different mechanisms of injury. CONCLUSIONS: It is important to include the mechanism of injury within intentional injuries because it provides different profiles of injury problems. Thus the simultaneous tabulation of injury mortality data by both intent and mechanism is a necessary step for identifying and prioritizing injury problems. The argument that good preventive measures could prevent both unintentional and intentional injuries was also confirmed. PMID- 11928981 TI - Comparison of two methods for assessing injuries among preschool children. AB - BACKGROUND: Most studies of injuries use health services records or recall rather than prospective methods, and there is no information on how these different methods compare. This study was aimed at comparing a report (retrospective) with a diary (prospective) for recording childhood injuries. METHODS: The study included 1,273 and 620 children, the retrospective and prospective subsamples, respectively, from a population based birth cohort in Pelotas, southern Brazil. The reported incidence of injuries in the preceding month were compared with those reported over month by diary (prospective study). RESULTS: Both methods were well accepted and 92.7% of the diaries were returned. One or more injuries per child month were reported for 20.8% (retrospective) and 48.4% (prospective) of the children. The total number of reported injuries for the 620 children were 145 (retrospective) and 715 (prospective). Using the prospective method as the gold standard, the retrospective method detected only 20.2% of all injuries. Under-reporting did not vary significantly with maternal education, but was greater (51.8%) for injuries requiring medical care than for those managed at home (18.3%; p=0.003). CONCLUSIONS: The diary was well accepted and resulted in higher incidences of reported injuries than the recall method, particularly for injuries that did not require medical care. Use of this method should be promoted to provide more complete epidemiological information to guide preventive strategies. PMID- 11928982 TI - Evaluating injury prevention interventions. PMID- 11928983 TI - Development and use of a population based injury surveillance system: the all Wales Injury Surveillance System (AWISS). AB - This report details the development and use of a population based emergency room surveillance system in the UK. Despite some difficulties in accessing high quality data the system has stimulated a considerable number of research and intervention projects. While surveillance systems with high quality data collection and coding parameters remain the gold standard, imperfect systems, particularly if population based, can play a substantial part in stimulating injury prevention initiatives. PMID- 11928984 TI - Genetic analysis of Group A rotaviruses: evidence for interspecies transmission of rotavirus genes. AB - Rotaviruses are the major cause of severe gastroenteritis in young children and animals. The rotavirus genome is composed of eleven segments of double-stranded RNA and can undergo genetic reassortment during mixed infections, leading to progeny viruses with novel or atypical phenotypes. There are numerous descriptions of rotavirus strains isolated from human and animals that share genetic and antigenic features of viruses from heterologous species. In many cases, genetic analysis by hybridization has clearly demonstrated the genetic relatedness of gene segments to those from viruses isolated from different species. Together with the observation that some virus strains appear to have been transmitted to a different species as a whole genome constellation, these data suggest that interspecies transmission occurs naturally, albeit at low frequencies. Although interspecies transmission has not been documented directly, there is an increasing number of reports of atypical rotaviruses that are apparently derived from transmission between: humans, cats and dogs; humans and cattle; humans and pigs; pigs and cattle; and pigs and horses. Interspecies evolutionary relationships are supported by phylogenetic analysis of rotavirus genes from different species. The emergence of novel strains derived from interspecies transmission has implications for the design and implementation of successful human rotavirus vaccine strategies. PMID- 11928985 TI - High level of sequence variation in the 3' noncoding region of Japanese encephalitis viruses isolated in Korea. AB - The 3' noncoding region (NCR) of Japanese encephalitis (JE) viruses isolated in Korea and Nakayama-NIH strain have been sequenced and compared with the 3' NCR sequences of other JE isolates reported previously. Sequence alignment of about 60 nucleotides (based on consensus sequence number) immediately downstream of the open reading frame (ORF) stop codon in the 3' NCR of the Korean isolates showed high degree of sequence variation and deletion; thus, this region was termed as the variable region. However, in the predicted RNA secondary structures, a similar type loop exists at the 5'-terminus of the 3' NCR of JE viruses, despite low level of sequence homology (22%) and deletion in the variable region. The phylogenetic tree based on the 3' NCR sequences of JE viruses including the variable region showed a similar pattern to that based on envelope genes; in that, there are two genetically different types of JE viruses in Korea. Therefore, the variable region would be a useful genetic marker for JE viruses. PMID- 11928986 TI - An AP-1 binding site mutation in HPV-16 LCR enhances E6/E7 promoter activity in human oral epithelial cells. AB - Expression of the human papillomavirus (HPV) E6 and E7 oncoproteins is responsible for the transforming ability of the virus. The HPV long control region (LCR) and E6/E7 promoter regulate transcription of the E6 and E7 viral oncogenes. However, factors involved in the stimulation of E6/E7 promoter activity in carcinogenesis are unclear. We previously identified a point mutation in an HPV-16 immortalized human oral keratinocyte cell line subsequently exposed to a tobacco-specific carcinogen. This mutation was located in the LCR at nucleotide 7633 and contains binding sites for the transcription activator, AP-1, overlapping with putative binding regions for the transcription factor, C/EBP, which represses the E6/E7 promoter. In this study, this mutation was analyzed by both electrophoretic mobility shift analysis and luciferase assays. We found that the point mutation enhanced the binding affinity of AP-1 to the LCR, thus stimulating the E6/E7 promoter activity. Our results suggest that mutations in binding sites for crucial regulators may be the result of exposure to carcinogens and could induce expression of the E6 and E7 oncogenes. PMID- 11928987 TI - Characterisation of transcripts from the human cytomegalovirus genes TRL7, UL20a, UL36, UL65, UL94, US3 and US34. AB - The genome of human cytomegalovirus (HCMV) has been studied extensively in some regions, but not others. In this study, transcripts of the genome were further characterised for open reading frames (ORFs) TRL7, UL36, UL65, UL94, US3 and US34, and for the previously unrecognised ORF, UL20a. Reverse transcription-PCR demonstrated the presence of spliced transcripts from the putative glycoprotein gene, UL20a, at early and late times post-infection. US3 full-length and spliced transcripts, including a previously unidentified transcript (US3ii), were described at immediate early times. Sequencing of the complete ORFs of UL20a and US3 from 21 clinical isolates showed that US3 is well conserved in all isolates (97-100% identity), whereas UL20a shows more variation at the nucleotide level, with 90-100% identity. The limits of transcription, and splice donor and acceptor sequences for UL20a and US3 were conserved in all isolates, indicating likely conservation of mRNA splicing patterns. Sequencing a late cDNA library identified the limits of transcription for ORFs TRL7, UL94 and US34 and transcription from the TRL7 ORF was confirmed by northern blotting. Transcripts were found that were congruent with UL36 and UL65, but these differed in the limits previously predicted for these ORFs. These findings show the variation between predicted and actual transcription and indicate the complex nature of transcription from HCMV ORFs. PMID- 11928988 TI - Varicella-zoster virus (VZV) mediates a delayed host shutoff independent of open reading frame (ORF) 17 expression. AB - Varicella-zoster virus (VZV) open reading frame 17 (ORF 17) is the gene corresponding to Herpes simplex-virus (HSV) UL41. The UL41 gene encodes the virion host shutoff factor (vhs), a RNase that has been the object of detailed studies. In contrast to HSV, knowledge about VZV mediated shutoff effects and the role of ORF 17 is poor. We investigated the ORF 17 expression in infected cells and analyzed shutoff effects. ORF 17 expression could not be proven in infected human fibroblast cell lines and melanoma (MeWo) cells. Only after induction by Phorbol 12-myristate 13-acetate an ORF 17 expression became detectable in MeWo cells. Nevertheless, using stable expressed GAPDH mRNA as a marker for mRNA degradation, a VZV mediated shutoff, independent of ORF 17 expression, became measurable. Transfection experiments demonstrated that transient ORF 17 expression did not decrease the cellular GAPDH mRNA level. We examined whether the VZV shutoff factor is a tegument protein causing an early shutoff or whether it needs to be expressed (delayed shutoff). The GAPDH mRNA level in Actinomycin D pretreated and infected MeWo cells did not decrease even faster than the theoretical decay rate based on a half-life of 24 h. These findings lead to the conclusion that the VZV shutoff factor is not a mature protein localized in the virion and that VZV causes a delayed virion host shutoff effect. PMID- 11928989 TI - Evolution of viruses by acquisition of genes that control nuclear functions in infected cells--an introduction. AB - The sequencing and deciphering of the human genome provided an insight into the gene complement of the human chromosomes as well as information on the nongenic sequences that constitute the chromosomal DNA molecules. The analyses of the genes and nongenic sequences in the human genome also provided important information on the presence of endogenous retroviruses, retroposons, retrotransposition of genes in the human genome as well as retroduplication of genes and distribution of the duplicated genes in different chromosomes. These issues were discussed in the first Special Issue of Virus Genes on Molecular Evolution of Viruses-Past and Present. In that issue, the discovery of the reverse transcriptase gene in archeabacteria, the retrovirus in drosophilae and endogenous retroviruses in the human genome were discussed. The aim of the present special issue on Molecular Evolution of Viruses is to consider the strategies developed by RNA and DNA viruses to control the nucleus and the nuclear functions in the infected cells. PMID- 11928990 TI - Sequence similarities and evolutionary relationships of influenza virus A hemagglutinins. AB - This study brings the analysis of amino acid sequences of hemagglutinin (HA) from the influenza virus A that can infect a wide variety of birds and mammals. 191 sequences belonging to all known 15 HA subtypes were compared. The emphasis was given on functional sites (receptor-binding cavity with its right and left edges) and degree of their conservation in each subtype. Three evolutionary trees of 15 avian HA representatives were constructed: one tree based on the alignment of the entire HA sequences and two trees based on the alignment of HA1 and HA2 chains, respectively. The results have shown that, despite low degree of sequence similarity among the 191 sequences of HA1 subunit, the active site is well conserved, and that there are only marginal differences in the clustering of the individual HA subtypes between the two subunit trees. In this respect, the subtype H9 seems to be the most fluctuating example. The proposals of the probable avian HAs that could be the closest relatives to human (mammalian) HAs were also provided for several HA subtypes. PMID- 11928992 TI - Effect of serum from chickens treated with clenbuterol on myosin accumulation, beta-adrenergic receptor population, and cyclic AMP synthesis in embryonic chicken skeletal muscle cell cultures. AB - Broiler chickens at 35 d of age were fed 1 ppm clenbuterol for 14 d. This level of dietary clenbuterol led to 5-7% increases in the weights of leg and breast muscle tissue. At the end of the 14-d period, serum was prepared from both control and clenbuterol-treated chickens, and was then employed as a component of cell culture media at a final concentration of 20% (v/v). Muscle cell cultures were prepared from both the leg and the breast muscle groups of 12-d chick embryos. Treatment groups included control chicken serum to which 10 nM, 50 nM, and 1 microM clenbuterol had been added, as well as cells grown in media containing 10% horse serum. Cultures were subjected to each treatment for 3 d, beginning on the seventh d in culture. Neither the percent fusion nor the number of nuclei in myotubes was significantly affected by any of the treatments. The quantity of myosin heavy chains (MHCs) was not increased by serum from clenbuterol-treated chickens in either breast or leg muscle cultures; however, the MHC quantity was 50-150% higher in cultures grown in control chicken serum to which 10 and 50 nM clenbuterol had also been added. The beta-adrenergic receptor (betaAR) population was 4000-7000 betaARs per cell in cultures grown in chicken serum, with leg muscle cultures having approximately 25-30% more receptors than breast muscle cultures. Receptor population was not significantly affected by the presence of clenbuterol or by the presence of serum from clenbuterol-treated chickens. In contrast, the betaAR population in leg and breast muscle cultures grown in the presence of 10% horse serum was 16,000-18,000 betaARs per cell. Basal concentration of cyclic adenosine 3':5'monophosphate (cAMP) was not significantly affected by the treatments. When cultures grown in chicken serum were stimulated for 10 min with 1 microM isoproterenol, limited increases of 12 20% in cAMP concentration above the basal levels were observed. However, when cultures grown in the presence of horse serum were stimulated with 1 microM isoproterenol, cAMP concentration was stimulated 5- to 9-fold above the basal levels. Thus, not only did cells grown in horse serum have a higher betaAR population, but also each receptor had a higher capacity for cAMP synthesis following isoproterenol stimulation. Finally, the hypothesis that clenbuterol exerts its action on muscle protein content by changes in cAMP concentration was tested. No correlation was apparent between basal cAMP concentration and MHC content. PMID- 11928993 TI - Activation of anchorage-independent growth of HT1080 human fibrosarcoma cells by dexamethasone. AB - Anchorage independence is an important hallmark of the transformation that correlates with tumorigenicity. We have isolated a variant clone of HT1080 human fibrosarcoma cells (cl-2) that is specifically defective in anchorage-independent growth. Interestingly, 10(-7) M dexamethasone (DEX) substantially rescued the anchorage-independent growth of cl-2 cells in semisolid culture. DEX also promoted the anchorage-independent growth of parental HT1080 cells. However, the agent had no effect on the anchorage-dependent growth of cl-2 and parental cells in ordinary liquid culture. Cell cycle analysis demonstrated that the population of G0/G1 cells increased, whereas that of S and G2/M cells decreased in growth arrested cl-2 cells in suspension culture. However, such an effect of anchorage loss on cell cycle progression was alleviated by adding 10(-7) M DEX. In cl-2 cells in semisolid culture, DEX suppressed the expression of P27Kip1, whereas it stimulated the expression of cyclin A and hyperphosphorylated retinoblastoma (Rb) proteins. On the other hand, DEX had no effect on cyclin D1 and P21Cap1 expression. These effects of DEX, except for the suppression of P27Kip1, were blocked by an antimicrofilament drug, cytochalasin D. Our results suggest that the stimulation of anchorage-independent growth by DEX involves at least two regulatory mechanisms, i.e., one that leads to the suppression of P27Kip1 protein without requiring cytoskeletal integrity, and another that requires cytoskeletal integrity, leading to stimulation of cyclin A and hyperphosphorylation of Rb protein. PMID- 11928991 TI - Sequence analysis of the Washington/1964 strain of human parainfluenza virus type 1 (HPIV1) and recovery and characterization of wild-type recombinant HPIV1 produced by reverse genetics. AB - A complete consensus sequence was determined for the genomic RNA of human parainfluenza virus type 1 (HPIV1) strain Washington/20993/1964 (HPIV1 WASH/64), a clinical isolate that previously was shown to be virulent in adults. The sequence exhibited a high degree of relatedness to both Sendai virus, a PIV1 virus recovered from mice, and human PIV3 (HPIV3) with regard to cis-acting regulatory regions and protein-coding sequences. This consensus sequence was used to generate a full-length antigenomic cDNA and to recover a recombinant wild-type HPIV1 (rHPIV1). Interestingly, the rHPIV1 could be rescued from full-length antigenomic rHPIV1 cDNA using HPIV3 support plasmids, HPIV1 support plasmids, or a mixture thereof. The replication of rHPIV1 in vitro and in the respiratory tract of hamsters was similar to that of its biologically derived parent virus. The similar biological properties of rHPIV1 and HPIV1 WASH/64 in vitro and in vivo, together with the previous demonstration of the virulence of this specific isolate in humans, authenticates the rHPIV1 sequence as that of a wild-type virus. This rHPIV1 can now be used to study the biological properties of HPIV1 and as a substrate to introduce attenuating mutations for the generation of live attenuated HPIV1 vaccine candidates. PMID- 11928994 TI - Loss of signal transduction and inhibition of lymphocyte locomotion in a ground based model of microgravity. AB - Inflammatory adherence to, and locomotion through the interstitium is an important component of the immune response. Conditions such as microgravity and modeled microgravity (MMG) severely inhibit lymphocyte locomotion in vitro through gelled type I collagen. We used the NASA rotating wall vessel bioreactor or slow-turning lateral vessel as a prototype for MMG in ground-based experiments. Previous experiments from our laboratory revealed that when lymphocytes (human peripheral blood mononuclear cells [PBMCs]) were first activated with phytohemaglutinin followed by exposure to MMG, locomotory capacity was not affected. In the present study, MMG inhibits lymphocyte locomotion in a manner similar to that observed in microgravity. Phorbol myristate acetate (PMA) treatment of PBMCs restored lost locomotory capacity by a maximum of 87%. Augmentation of cellular calcium flux with ionomycin had no restorative effect. Treatment of lymphocytes with mitomycin C prior to exposure to MMG, followed by PMA, restored locomotion to the same extent as when nonmitomycin C-treated lymphocytes were exposed to MMG (80-87%), suggesting that deoxyribonucleic acid replication is not essential for the restoration of locomotion. Thus, direct activation of protein kinase C (PKC) with PMA was effective in restoring locomotion in MMG comparable to the normal levels seen in Ig cultures. Therefore, in MMG, lymphocyte calcium signaling pathways were functional, with defects occurring at either the level of PKC or upstream of PKC. PMID- 11928995 TI - Delivery and storage of single embryos, sperm, or cells in microglass capillaries. PMID- 11928996 TI - Effects of bone marrow stromal cells on the structural and functional polarity of primary rat hepatocytes. PMID- 11928997 TI - Preparation of isolated human muscle fibers: a technical report. AB - The aim of this study was to develop a technique to culture satellite cells from isolated intact fast or slow human muscle fibers. Previous studies have been carried out on small rodent muscles where the fibers run from tendon to tendon, but this is the first description of the modification of this technique for much larger human muscles. We have demonstrated that the human muscle fibers are in fact segmental, and we have also shown that it is possible to obtain very pure satellite cell cultures. We discuss the importance of this technique as a source of highly purified muscle cell cultures, which can be used for further studies on satellite cell behavior. PMID- 11928998 TI - Characterization of cell lines developed from the Colorado potato beetle, Leptinotarsa decemlineata Say (Coleoptera: Chrysomelidae). AB - In order to isolate new pathogens (viruses, microsporidia, etc.) or to evaluate the efficiency of some pathogens (serovarieties and mutants of Bacillus thuringiensis, fungi, etc.) in the control of Colorado potato beetle, an economically important pest, we established four cell lines from tissues of this insect. One was initiated from embryonated egg fragments in the M3 medium supplemented with 20% fetal bovine serum (FBS) and then transferred after several passages to the Ex-Cell 400 medium with 20% FBS. Another was initiated from larval hemocytes in Ex-Cell 400 with 5% FBS. Finally, two other cell lines were initiated from adult hemocytes: one in the Ex-Cell 400 with 20% FBS and 1% of lipid mixture and the other in the Ex-Cell 400 with 5% FBS only. These cell lines have been characterized by their morphology with light and electron microscopy, their karyotypes, cell growth, and isozyme analysis. Each cell line differed in morphologic, karyologic, growth, and isozyme patterns. The cell line initiated from embryonated eggs was growing slower than the three initiated from hemocytes. The cytotoxicity of solubilized crystal delta-endotoxins from different B. thuringiensis formulations (M-One, Trident, MYX-1806, Teknar-HPD, and Thuricide) and of destruxins, mycotoxins from Metarhizium anisopliae, was tested on these cell lines. They are sensitive to the solubilized toxins of some strains of B. thuringiensis (serovar. San Diego and serovar. tenebrionis) and to destruxins, and they can be used for the bioassay and detection of toxins and for the study of the mechanism of their action on coleopteran cells. PMID- 11928999 TI - Comparative PCR analysis for detection of mycoplasma infections in continuous cell lines. AB - Mycoplasma contamination of cell lines is one of the major problems in cell culturing. About 15-35% of all cell lines are infected with a limited number of mycoplasma species of predominantly human, swine, or bovine origin. We examined the mycoplasma contamination status in 495 cell cultures by polymerase chain reaction (PCR) assay, microbiological culture method, and deoxyribonucleic acid ribonucleic acid (DNA-RNA) hybridization, and in 103 cell cultures by PCR and DNA RNA hybridization, in order to determine the sensitivity and specificity of the PCR assay in routine cell culture. For those two cohorts, results for the three or two assays were concordant in 92 and 91% of the cases, respectively. The sensitivity (detection of true positives) of this PCR detection assay was 86%, and the specificity (detection of true negatives) was 93%, with positive and negative predictive values (probability of correct results) of 73 and 97%, respectively. PCR defined the mycoplasma status with 92% accuracy (detection of true positives and true negatives). The mycoplasma contaminants were speciated by analyzing the PCR amplification fragment using several restriction enzymes. Most of the cultures (47%) were infected with Mycoplasma fermentans, followed by M. hyorhinis (19%), M. orale (10%), M. arginini (9%), Acholeplasma laidlawii (6%), and M. hominis (3%). To sum up, PCR represents a sensitive, specific, accurate, inexpensive, and quick mycoplasma detection assay that is suitable for the routine screening of cell cultures. PMID- 11929000 TI - Comparative antibiotic eradication of mycoplasma infections from continuous cell lines. AB - Accumulating data implicate mycoplasma contamination as the single biggest problem in the culture of continuous cell lines. Mycoplasma infection can affect virtually every parameter and functional activity of the eukaryotic cells. A successful alternative to discarding infected cultures is to attempt to eliminate the contaminants by treatment with specific and efficient antimycoplasma antibiotics. The addition of antibiotics to the culture medium during a limited period of time (1-3 wk) is a simple, inexpensive, and very practical approach for decontaminating continuous cell lines. Here, we examined the effectiveness of several antibiotic treatment protocols that we have employed routinely in our cell lines bank. On an aggregate, 673 cultures from 236 chronically mycoplasma positive cell lines were exposed to one of the following five antibiotic regimens: mycoplasma removal agent (quinolone; a 1-wk treatment), enrofloxacin (quinolone; 1 wk), sparfloxacin (quinolone; 1 wk), ciprofloxacin (quinolone; 2 wk), and BM-Cyclin (alternating tiamulin and minocycline; 3 wk). The mycoplasma infection was permanently (as determined by three solid mycoplasma detection assays) eliminated by the various antibiotics in 66-85% of the cultures treated. Mycoplasma resistance was seen in 7-21%, and loss of the culture as a result of cytotoxically caused cell death occurred in 3-11% of the cultures treated. Overall, 223 of the 236 mycoplasma-positive cell lines could be cured in a first round of antibiotic treatment with at least one regimen. Taken together, 95% of the mycoplasma-infected cell lines were permanently cleansed of the contaminants by antibiotic treatment, which validates this approach as an efficient and technically simple mycoplasma eradication method. PMID- 11929001 TI - Characterization and efficacy of PKH26 as a probe to study the replication history of the human hematopoietic KG1a progenitor cell line. AB - The PKH26 dye can, in principle, be used for the study of asymmetric cell divisions (ASDs). A requirement for the identification of ASDs based on fluorescence intensity is that the PKH26 dye is distributed equally between daughter cells at each division, but this has not been demonstrated at a single cell level. The efficacy of PKH26 as a probe for the study of ASDs was examined using the human hematopoietic KG1a cell. An automated time-lapse fluorescent microscope system was used to determine changes in cell size and fluorescence intensity during culture, and track cell divisions. The images of daughter cells were analyzed using the Isee software to determine the distribution of PKH26 dye between daughter cells. Ratios of cell size, mean fluorescence intensity, and total fluorescence intensity were calculated by dividing the values for one daughter cell by the value of the other daughter cell. The ratios for cell size, mean intensity, and total intensity were 1.13 +/- 0.12, 1.08 +/- 0.07, and 1.15 +/- 0.14 (mean +/- SD), respectively. Thus, PKH26 is not distributed equally to both daughter cells upon cell division. However, the replication history of individual KG1a cells can be reliably deduced for up to three divisions based solely on the mean and total fluorescence intensity of the PKH26 dye, using PKH26 concentrations below the chemical and phototoxic limits (2 microM). PMID- 11929002 TI - An improved method for the collagen gel contraction assay. AB - The collagen gel contraction (CGC) assay is used frequently to study the cell mediated reorganization of the extracellular natrix. In a typical CGC assay, cells embedded in a disk-shaped lattice (gel) of native type I collagen fibers compress the fibers and, consequently, reduce the diameter of the collagen disk within h or d. The degree to which the collagen is contracted is usually quantified by measurement of the diameter or the area of the disk. During CCC assays, friction or adhesion (or both) between gels and their culture containers can cause gels to be incompletely contracted or to acquire distorted shapes. Such occurrences degrade the reproducibility and reliability of measurements of gel dimensions. To address these problems, we developed an oil-supported collagen retraction (OSCR) assay that creates an environment of low friction and adhesion around the contracting collagen gel. The OSCR assay is accomplished with simple equipment and is easily performed, sensitive, and consistently yields fully contracted gels with minimal distortion. PMID- 11929003 TI - Impact of Internet is huge. PMID- 11929004 TI - Do as I say not as I do. PMID- 11929005 TI - Harvesting the fruits of research: new guidelines on nutrition and physical activity. PMID- 11929006 TI - The mammographic screening trials: commentary on the recent work by Olsen and Gotzsche. PMID- 11929007 TI - Cancer anorexia-cachexia syndrome: current issues in research and management. AB - Cachexia is among the most debilitating and life-threatening aspects of cancer. Associated with anorexia, fat and muscle tissue wasting, psychological distress, and a lower quality of life, cachexia arises from a complex interaction between the cancer and the host. This process includes cytokine production, release of lipid-mobilizing and proteolysis-inducing factors, and alterations in intermediary metabolism. Cachexia should be suspected in patients with cancer if an involuntary weight loss of greater than five percent of premorbid weight occurs within a six-month period. The two major options for pharmacological therapy have been either progestational agents, such as megestrol acetate, or corticosteroids. However, knowledge of the mechanisms of cancer anorexia-cachexia syndrome has led to, and continues to lead to, effective therapeutic interventions for several aspects of the syndrome. These include antiserotonergic drugs, gastroprokinetic agents, branched-chain amino acids, eicosapentanoic acid, cannabinoids, melatonin, and thalidomide--all of which act on the feeding regulatory circuitry to increase appetite and inhibit tumor-derived catabolic factors to antagonize tissue wasting and/or host cytokine release. Because weight loss shortens the survival time of cancer patients and decreases performance status, effective therapy would extend patient survival and improve quality of life. PMID- 11929008 TI - American Cancer Society guidelines on nutrition and physical activity for cancer prevention: Reducing the risk of cancer with healthy food choices and physical activity. AB - The American Cancer Society (ACS) has set aggressive challenge goals for the nation to decrease cancer incidence and mortality--and to improve the quality of life of cancer survivors--by the year 2015. To address these critical goals, the ACS publishes the Nutrition and Physical Activity Guidelines to serve as a foundation for its communication, policy, and community strategies and ultimately, to affect dietary and physical activity patterns among Americans. These guidelines, published every five years, are developed by a national panel of experts in cancer research, prevention, epidemiology, public health, and policy, and as such, they represent the most current scientific evidence related to dietary and activity patterns and cancer risk. The American Cancer Society guidelines include recommendations for individual choices regarding diet and physical activity patterns, but those choices occur within a community context that either facilitates or interferes with healthy behaviors. Therefore, this committee presents one key recommendation for community action to accompany the four recommendations for individual choices for nutrition and physical activity to reduce cancer risk. This recommendation for community action underscores just how important community measures are to the support of healthy behaviors by means of increasing access to healthful food choices and opportunities to be physically active. The ACS guidelines are consistent with guidelines from the American Heart Association for the prevention of coronary heart disease as well as for general health promotion, as defined by the Department of Health and Human Services' 2000 Dietary Guidelines for Americans. PMID- 11929009 TI - A quality and efficiency analysis of the IMFAST segmentation algorithm in head and neck "step & shoot" IMRT treatments. AB - The performance of segmentation algorithms used in IMFAST for "step & shoot" IMRT treatment delivery is evaluated for three head and neck clinical treatments of different optimization objectives. The segmentation uses the intensity maps generated by the in-house TPS PLANUNC using the index-dose minimization algorithm. The dose optimization objectives include PTV dose uniformity and dose volume histogram-specified critical structure sparing. The optimized continuous intensity maps were truncated into five and ten intensity levels and exported to IMFAST for MLC segments optimization. The MLC segments were imported back to PLUNC for dose optimization quality calculation. The five basic segmentation algorithms included in IMFAST were evaluated alone and in combination with either tongue and groove/match line correction or fluence correction or both. Two criteria were used in the evaluation: treatment efficiency represented by the total number of MLC segments and optimization quality represented by a clinically relevant optimization quality factor. We found that the treatment efficiency depends first on the number of intensity levels used in the intensity map and second the segmentation technique used. The standard optimal segmentation with fluence correction is a consistent good performer for all treatment plans studied. All segmentation techniques evaluated produced treatments with similar dose optimization quality values, especially when ten-level intensity maps are used. PMID- 11929010 TI - Comparing calibration methods of electron beams using plane-parallel chambers with absorbed-dose to water based protocols. AB - Recent absorbed-dose-based protocols allow for two methods of calibrating electron beams using plane-parallel chambers, one using the N(Co)D,w for a plane parallel chamber, and the other relying on cross-calibration of the plane parallel chamber in a high-energy electron beam against a cylindrical chamber which has an N(Co)D,w factor. The second method is recommended as it avoids problems associated with the Pwall correction factors at 60Co for plane-parallel chambers which are used in the determination of the beam quality conversion factors. In this article we investigate the consistency of these two methods for the PTW Roos, Scanditronics NACP02, and PTW Markus chambers. We processed our data using both the AAPM TG-51 and the IAEA TRS-398 protocols. Wall correction factors in 60Co beams and absorbed-dose beam quality conversion factors for 20 MeV electrons were derived for these chambers by cross-calibration against a cylindrical ionization chamber. Systematic differences of up to 1.6% were found between our values of Pwall and those from the Monte Carlo calculations underlying AAPM TG-51, and up to 0.6% when comparing with the IAEA TRS-398 protocol. The differences in Pwall translate directly into differences in the beam quality conversion factors in the respective protocols. The relatively large spread in the experimental data of Pwall, and consequently the absorbed-dose beam quality conversion factor, confirms the importance of the cross-calibration technique when using plane-parallel chambers for calibrating clinical electron beams. We confirmed that for well-guarded plane-parallel chambers, the fluence perturbation correction factor at d(max) is not significantly different from the value at d(ref). For the PTW Markus chamber the variation in the latter factor is consistent with published fits relating it to average energy at depth. PMID- 11929011 TI - Optimizing the shape of ultrasound transducers for interstitial thermal ablation. AB - Heat deposition by interstitial routes, especially with ultrasound-based instruments, is becoming a valuable therapeutic option for the treatments of sites, which are difficult to access from outside of the body. The active part of most interstitial ultrasound applicators described in the literature is logically tubular to induce cylindrical volumes of coagulation necrosis. Because the pressure generated by such tubular transducers falls off rapidly with radial distance, we previously proposed using a rotating plane transducer. For a plane wave, the pressure fall-off is only due to attenuation, which makes deeper lesions and shorter treatment times possible. This work represents an advance in the development of ultrasound applicators designed for interstitial applications. This new applicator used a rotating slightly focused transducer. A brief theoretical analysis resulted in the choice of a long focal distance of 22 mm to obtain a nearly constant pressure all along the therapeutic depth. To experimentally validate this focal distance, pressure measurements were made in a tissue mimicking liquid phantom and the results were compared with those obtained with a plane transducer. In vitro experiments showed that necrosis could be induced at a depth of 15 mm. In the same conditions, the greatest depth attained with a plane transducer was only 10 mm. Because each individual lesion is narrower, more lesions and more time are required to necrose a cylindrical volume. The main advantage of this new type of applicator is that it can be used to induce necrosis at a greater depth without varying either the frequency, the intensity or the transducer cooling efficiency. PMID- 11929012 TI - Noise aliasing in interline-video-based fluoroscopy systems. AB - Video-based imaging systems for continuous (nonpulsed) x-ray fluoroscopy use a variety of video formats. Conventional video-camera systems may operate in either interlaced or progressive-scan modes, and CCD systems may operate in interline- or frame-transfer modes. A theoretical model of the image noise power spectrum corresponding to these formats is described. It is shown that with respect to frame-transfer or progressive-readout modes, interline or interlaced cameras operating in a frame-integration mode will result in a spectral shift of 25% of the total image noise power from low spatial frequencies to high. In a field integration mode, noise power is doubled with most of the increase occurring at high spatial frequencies. The differences are due primarily to the effect of noise aliasing. In interline or interlaced formats, alternate lines are obtained with each video field resulting in a vertical sampling frequency for noise that is one half of the physical sampling frequency. The extent of noise aliasing is modified by differences in the statistical correlations between video fields in the different modes. The theoretical model is validated with experiments using an x-ray image intensifier and CCD-camera system. It is shown that different video modes affect the shape of the noise-power spectrum and therefore the detective quantum efficiency. While the effect on observer performance is not addressed, it is concluded that in order to minimize image noise at the critical mid-to-high spatial frequencies for a specified x-ray exposure, fluoroscopic systems should use only frame-transfer (CCD camera) or progressive-scan (conventional video) formats. PMID- 11929013 TI - Evaluation of layer decomposition for multiframe quantitative coronary angiography. AB - Multiframe quantitative coronary angiography is typically performed by averaging measurements of artery diameter over multiple frames. This approach reduces errors attributable to random noise but may not reduce systematic errors caused by background structures, nonlinear system response, and motion blur. We attempt to reduce these sources of error by decomposing the image sequence into moving layers, one of which includes the artery. We embed simulated arteries into clinical angiographic sequences so that the true vessel dimensions are known accurately. The measurement tasks are minimum diameter, geometric percent stenosis, and densitometric percent stenosis. We compare measurements for single and multiple raw images, single images with fixed mask subtraction, single and multiple images with layered background subtraction, and time-averaged layer images. We find that both multiframe averaging and layer decomposition significantly improve geometric and densitometric accuracy compared with single frame measurements. The best results were obtained by averaging measurements from multiple frames of layered background-subtracted images. PMID- 11929014 TI - The x-ray sensitivity of amorphous selenium for mammography. AB - A study of the x-ray sensitivity of amorphous selenium (a-Se) for digital mammography has been performed. A uniform layer of a-Se was deposited on a glass substrate with electrodes on both surfaces. The deposition procedure was identical to that used for a-Se flat-panel detectors. A high voltage was applied to the top surface of the a-Se layer in order to establish an electric field E(Se). Then the sample was exposed to x rays with 27 kVp spectra generated from an x-ray tube with a molybdenum (Mo) target. The mean x-ray energy of the spectrum used was approximately 16.6 keV. The x-ray current generated by the a-Se layer was measured as a function of E(Se). From the current measurement and the estimation of total x-ray energy absorbed in the a-Se, the energy required to create one electron-hole pair (EHP), W, was determined as a function of E(Se). It was found that at the most commonly used E(Se) of 10 V/microm, W was measured as 64 eV. This is considerably higher than the widely accepted typical value of W = 50 eV measured at higher x-ray photon energies (e.g., 50 keV). The dependence of W as a function of E(Se) can be best fitted using the empirical expression of E(Se)-gamma. This relationship is consistent with the results obtained at higher x-ray energies. This article provides an accurate measurement of x-ray sensitivity of a-Se at mammographic energies independent of detector operation, such as the most recently developed flat-panel detectors. The results will be a useful tool for investigation and optimization of a-Se-based x-ray imaging detectors, such as determination of pixel fill-factor and optimal E(Se) during operation. PMID- 11929015 TI - Accurate localization of intracavitary brachytherapy applicators from 3D CT imaging studies. AB - PURPOSE: To present an accurate method to identify the positions and orientations of intracavitary (ICT) brachytherapy applicators imaged in 3D CT scans, in support of Monte Carlo photon-transport simulations, enabling accurate dose modeling in the presence of applicator shielding and interapplicator attenuation. MATERIALS AND METHODS: The method consists of finding the transformation that maximizes the coincidence between the known 3D shapes of each applicator component (colpostats and tandem) with the volume defined by contours of the corresponding surface on each CT slice. We use this technique to localize Fletcher-Suit CT-compatible applicators for three cervix cancer patients using post-implant CT examinations (3 mm slice thickness and separation). Dose distributions in 1-to-1 registration with the underlying CT anatomy are derived from 3D Monte Carlo photon-transport simulations incorporating each applicator's internal geometry (source encapsulation, high-density shields, and applicator body) oriented in relation to the dose matrix according to the measured localization transformations. The precision and accuracy of our localization method are assessed using CT scans, in which the positions and orientations of dense rods and spheres (in a precision-machined phantom) were measured at various orientations relative to the gantry. RESULTS: Using this method, we register 3D Monte Carlo dose calculations directly onto post insertion patient CT studies. Using CT studies of a precisely machined phantom, the absolute accuracy of the method was found to be +/-0.2 mm in plane, and +/-0.3 mm in the axial direction while its precision was +/-0.2 mm in plane, and +/-0.2 mm axially. CONCLUSION: We have developed a novel, and accurate technique to localize intracavitary brachytherapy applicators in 3D CT imaging studies, which supports 3D dose planning involving detailed 3D Monte Carlo dose calculations, modeling source positions, shielding and interapplicator shielding, accurately. PMID- 11929016 TI - Fiducial-based targeting accuracy for external-beam radiotherapy. AB - The accuracy of fiducial-based alignment of external radiotherapy beams is analyzed. The study considers three basic computational methods to determine the target position--the exact closed-form solution for three fiducials, the solution via singular value decomposition for four or more fiducials, and the iterative solution for any number of fiducials--and assesses their accuracy, robustness, and efficiency. Particular attention is paid to inaccuracies arising from the variability of fiducial positions in soft tissue. In nearly every test case it is found that all three solution methods, when properly implemented, yield the same result for the target position, but that the method of singular value decomposition must be modified to distinguish rotations from reflections. When an accurate measure of the rotation of the target site is needed, four fiducials give much better results than three, while more than five fiducials gain little further improvement. PMID- 11929017 TI - Introduction of audio gating to further reduce organ motion in breathing synchronized radiotherapy. AB - With breathing synchronized radiotherapy (BSRT), a voltage signal derived from an organ displacement detector is usually displayed on the vertical axis whereas the elapsed time is shown on the horizontal axis. The voltage gate window is set on the breathing voltage signal. Whenever the breathing signal falls between the two gate levels, a gate pulse is produced to enable the treatment machine. In this paper a new gating mechanism, audio (or time-sequence) gating, is introduced and is integrated into the existing voltage gating system. The audio gating takes advantage of the repetitive nature of the breathing signal when repetitive audio instruction is given to the patient. The audio gating is aimed at removing the regions of sharp rises and falls in the breathing signal that cannot be removed by the voltage gating. When the breathing signal falls between voltage gate levels as well as between audio-gate levels, the voltage- and audio-gated radiotherapy (ART) system will generate an AND gate pulse. When this gate pulse is received by a linear accelerator, the linear accelerator becomes "enabled" for beam delivery and will deliver the beam when all other interlocks are removed. This paper describes a new gating mechanism and a method of recording beam-on signal, both of which are, configured into a laptop computer. The paper also presents evidence of some clinical advantages achieved with the ART system. PMID- 11929018 TI - Film dosimetry for intensity modulated radiation therapy: dosimetric evaluation. AB - X-ray film has been used for the dosimetry of intensity modulated radiation therapy (IMRT). However, the over-response of the film to low-energy photons is a significant problem in photon beam dosimetry, especially in regions outside penumbra. In IMRT, the radiation field consists of multiple small fields and their outside-penumbra regions; thus, the film dosimetry, for it involves the source of over-response in its radiation field. In this study we aim to verify and possibly improve film dosimetry for IMRT. Two types of modulated beams were constructed by combining five to seven different static radiation fields using 6 MV x rays. For verifying film dosimetry, x-ray films and an ion chamber were used to measure dose profiles at various depths in a phantom. The film setups include both parallel and perpendicular arrangements against the beam incident direction. In addition, to reduce an over-response, we placed 0.01 in. (0.25 mm) thick lead filters on both sides of the film. Compared with ion-chamber measurement, measured dose profiles showed the film over-response at outside-penumbra and low dose regions. The error increased with depths and approached 15% as a maximum for the field size of 15 cm x 15 cm at 10 cm depth. The use of filters reduced the error down to 3%. In this study we demonstrated that film dosimetry for IMRT involves sources of error due to its over-response to low-energy photons, with the error most transparent in the low-dose region. The use of filters could enhance the accuracy in film dosimetry for IMRT. In this regard, the use of an optimal filter condition is recommended. PMID- 11929019 TI - The use of quantitative temperature images to predict the optimal power for focused ultrasound surgery: in vivo verification in rabbit muscle and brain. AB - In this study, we investigated the use of MRI-derived thermal imaging for determining the exposure parameters for focused ultrasound (FUS) surgery. Since the temperature rise induced by a FUS beam scales linearly with power, the temperature maps acquired during subthreshold sonications can be used to determine the power necessary to produce thermal tissue damage with a desired size. Thermal images acquired during multiple sonications delivered at different locations in rabbit thigh muscle and brain tissue in vivo were analyzed to test this hypothesis. First, the linearity of the induced temperature rise with the acoustic power was tested. Next, the temperature maps acquired during preliminary low power sonications were scaled up until the estimated size of the tissue damage was equal to the tissue damage size of subsequent high power sonications. A threshold thermal dose was used to estimate the onset of thermal damage. The predicted power (based on amount of scaling required to reach the target size) was then compared to the true high power value. Overall, the temperature rise varied linearly with power (slope of deltaThigh/deltaTlow vs Power(high)/Power(low) = 0.97, 0.93 for pairs of sonications at each location in brain, muscle). The predicted power matched the true high power in the brain sonications (slope = 1.04). The predicted power underestimated the true high power in the muscle sonications (slope = 0.87). This under-prediction was due to a deviation from linearity in those cases where tissue damage was detected in subsequent MR images (slope of deltaThigh/deltaTlow vs Power(high)/Power(low) = 1.02, 0.84 for no tissue damage, tissue damage). The source of this deviation was not clear from these experiments. Even with this underestimation of the power, this method will be useful because it will allow an estimate of the proper power to use during FUS surgery without exact knowledge of the tissue parameters. PMID- 11929020 TI - Effect of respiratory gating on reducing lung motion artifacts in PET imaging of lung cancer. AB - Positron emission tomography (PET) has shown an increase in both sensitivity and specificity over computed tomography (CT) in lung cancer. However, motion artifacts in the 18F fluorodioxydoglucose (FDG) PET images caused by respiration persists to be an important factor in degrading PET image quality and quantification. Motion artifacts lead to two major effects: First, it affects the accuracy of quantitation, producing a reduction of the measured standard uptake value (SUV). Second, the apparent lesion volume is overestimated. Both impact upon the usage of PET images for radiation treatment planning. The first affects the visibility, or contrast, of the lesion. The second results in an increase in the planning target volume, and consequently a greater radiation dose to the normal tissues. One way to compensate for this effect is by applying a multiple frame capture technique. The PET data are then acquired in synchronization with the respiratory motion. Reduction in smearing due to gating was investigated in both phantoms and patient studies. Phantom studies showed a dependence of the reduction in smearing on the lesion size, the motion amplitude, and the number of bins used for data acquisition. These studies also showed an improvement in the target-to-background ratio, and a more accurate measurement of the SUV. When applied to one patient, respiratory gating showed a 28% reduction in the total lesion volume, and a 56.5% increase in the SUV. This study was conducted as a proof of principle that a gating technique can effectively reduce motion artifacts in PET image acquisition. PMID- 11929021 TI - Parker weights revisited. AB - The short-scan case in fan-beam computed tomography requires the introduction of a weighting function to handle redundant data. Parker introduced such a weighting function for a scan over pi plus the opening angle of the fan. In this article we derive a general class of weighting functions for arbitrary scan angles between pi plus fan angle and 2pi (over-scan). These weighting functions lead to mathematically exact reconstructions in the continuous case. Parker weights are a special case of a weighting function that belongs to this class. It will be shown that Parker weights are not generally the best choice in terms of noise reduction, especially when there is considerable over-scan. We derive a new weighting function that has a value of 0.5 for most of the redundant data and is smooth at the boundaries. PMID- 11929022 TI - Neural network based automated algorithm to identify joint locations on hand/wrist radiographs for arthritis assessment. AB - Arthritis is a significant and costly healthcare problem that requires objective and quantifiable methods to evaluate its progression. Here we describe software that can automatically determine the locations of seven joints in the proximal hand and wrist that demonstrate arthritic changes. These are the five carpometacarpal (CMC1, CMC2, CMC3, CMC4, CMC5), radiocarpal (RC), and the scaphocapitate (SC) joints. The algorithm was based on an artificial neural network (ANN) that was trained using independent sets of digitized hand radiographs and manually identified joint locations. The algorithm used landmarks determined automatically by software developed in our previous work as starting points. Other than requiring user input of the location of nonanatomical structures and the orientation of the hand on the film, the procedure was fully automated. The software was tested on two datasets: 50 digitized hand radiographs from patients participating in a large clinical study, and 60 from subjects participating in arthritis research studies and who had mild to moderate rheumatoid arthritis (RA). It was evaluated by a comparison to joint locations determined by a trained radiologist using manual tracing. The success rate for determining the CMC, RC, and SC joints was 87%-99%, for normal hands and 81%-99% for RA hands. This is a first step in performing an automated computer-aided assessment of wrist joints for arthritis progression. The software provides landmarks that will be used by subsequent image processing routines to analyze each joint individually for structural changes such as erosions and joint space narrowing. PMID- 11929023 TI - Communication and sampling rate limitations in IMRT delivery with a dynamic multileaf collimator system. AB - The delivery of an intensity modulated radiation field with a dynamic multileaf collimator (MLC) requires precise correlation between MLC positions and cumulative monitor units (MUs). The purpose of this study is to investigate the precision of this correlation as a function of delivered MUs and dose rate. A semi-Gaussian shaped intensity profile and a simple geometric intensity pattern consisting of four square segments were designed to deliver a total of 1, 4, 16, 64, and 100 MUs at three different dose rates of 100, 400, and 600 MU/min. The semi-Gaussian intensity pattern was delivered using both sliding window and step and shoot techniques. The dose profiles of this intensity pattern were measured with films. The four square intensity pattern was delivered using step and shoot and conventional delivery techniques for comparison. Because of geometrical symmetry, the dose to each segment in this intensity pattern is expected to be the same when the same MU is assigned to each segment. An ionization chamber was used to measure the dose in the center of each of the four square segments. For the semi-Gaussian shaped profile, significant artifacts were observed when the profile was delivered with small MUs and/or at a high dose rate. For the four square intensity pattern, the dose measured in each segment presented a large variation when delivered with small MUs and a high dose rate. The variation increases as the MU/segment decreases and as the dose rate increases. These MU and dose rate dependencies were not observed when the intensity pattern was delivered using a conventional delivery technique. The observed distortion of the semi-Gaussian profile and dose variations among the segments of the four square intensity pattern are explained by considering the sampling rate and the communication time lag between the control systems. Finally, clinical significance is discussed. PMID- 11929024 TI - Brachytherapy dose-volume histogram computations using optimized stratified sampling methods. AB - A stratified sampling method for the efficient repeated computation of dose volume histograms (DVHs) in brachytherapy is presented as used for anatomy based brachytherapy optimization methods. The aim of the method is to reduce the number of sampling points required for the calculation of DVHs for the body and the PTV. From the DVHs are derived the quantities such as Conformity Index COIN and COIN integrals. This is achieved by using partial uniform distributed sampling points with a density in each region obtained from a survey of the gradients or the variance of the dose distribution in these regions. The shape of the sampling regions is adapted to the patient anatomy and the shape and size of the implant. For the application of this method a single preprocessing step is necessary which requires only a few seconds. Ten clinical implants were used to study the appropriate number of sampling points, given a required accuracy for quantities such as cumulative DVHs, COIN indices and COIN integrals. We found that DVHs of very large tissue volumes surrounding the PTV, and also COIN distributions, can be obtained using a factor of 5-10 times smaller the number of sampling points in comparison with uniform distributed points. PMID- 11929025 TI - Measurement of absorbed dose with a bone-equivalent extrapolation chamber. AB - A hybrid phantom-embedded extrapolation chamber (PEEC) made of Solid Water and bone-equivalent material was used for determining absorbed dose in a bone equivalent phantom irradiated with clinical radiation beams (cobalt-60 gamma rays; 6 and 18 MV x rays; and 9 and 15 MeV electrons). The dose was determined with the Spencer-Attix cavity theory, using ionization gradient measurements and an indirect determination of the chamber air-mass through measurements of chamber capacitance. The collected charge was corrected for ionic recombination and diffusion in the chamber air volume following the standard two-voltage technique. Due to the hybrid chamber design, correction factors accounting for scatter deficit and electrode composition were determined and applied in the dose equation to obtain absorbed dose in bone for the equivalent homogeneous bone phantom. Correction factors for graphite electrodes were calculated with Monte Carlo techniques and the calculated results were verified through relative air cavity dose measurements for three different polarizing electrode materials: graphite, steel, and brass in conjunction with a graphite collecting electrode. Scatter deficit, due mainly to loss of lateral scatter in the hybrid chamber, reduces the dose to the air cavity in the hybrid PEEC in comparison with full bone PEEC by 0.7% to approximately 2% depending on beam quality and energy. In megavoltage photon and electron beams, graphite electrodes do not affect the dose measurement in the Solid Water PEEC but decrease the cavity dose by up to 5% in the bone-equivalent PEEC even for very thin graphite electrodes (<0.0025 cm). In conjunction with appropriate correction factors determined with Monte Carlo techniques, the uncalibrated hybrid PEEC can be used for measuring absorbed dose in bone material to within 2% for high-energy photon and electron beams. PMID- 11929026 TI - Point/counterpoint. Medical physicists should not publicly endorse commercial products. PMID- 11929028 TI - Diagnostic modalities for gastroesophageal reflux. AB - OBJECTIVE: To evaluate commonly utilized diagnostic modalities to detecting Gastroesophageal Reflux (GER). METHODS: Sixty children aged 1-72 months (mean age 14.7 months) with symptoms suggestive of Gastroesosphageal Reflux (GER) were investigated and subjected to upper gastrointestinal endoscopy and esophageal biopsy (EB), gastroesophageal scintiscanning (GS) and 24 hour ambulatory pH monitoring. RESULT: GER was detected in 28 (46.7%) cases by one or more diagnostic modalities. Ambulatory 24 hour pH monitoring was positive in higher proportion (43.3%) of cases in comparison to other modalities, followed by EB (38.3%) and GS (30%). Considering 24 hour pH monitoring as the gold standard, esophageal biopsy was positive in 22/26 cases (84.6%) detected by 24 hour pH monitoring with a specificity of 97.1% as compared to 17/26 cases (65.4%) by gastroesophageal scintiscanning with a specificity of 97.1%. When compared with EB results, amongst various parameters measured during 24 hour pH monitoring, Reflux index (RI) ranked highest (sensitivity 95.6 % and specificity 89.2 %) followed by duration of longest episode > 20 minutes and Euler Byrne score. Oscillatory index, calculated from tracings of pH monitoring, even though ranked lower because of its low sensitivity helped to pick up 2 cases missed by EB and RI. CONCLUSION: Our results suggest that a combination of diagnostic modalities may be required to diagnose GER in young children. Ambulatory 24 hour pH monitoring appears to be the single best investigation and combining it with EB and/or GS can help to detect maximum number of cases. PMID- 11929029 TI - Gastroesophageal reflux as cause of chronic respiratory symptoms. AB - OBJECTIVE: Gastroesophageal reflux (GER) is a relatively common disorder in infants and children. It maybe associated with severe complications. The coexistence of GER and a wide range of respiratory symptoms has been reported. The purpose of our study was to investigate the relationship between chronic respiratory symptoms and GERD as an underlying cause. To our knowledge, there is not a method known study for identifying this relationship and prevalence in our area. METHODS: The study group consists of fifty-two (4 months-10 years) children who were referred to pediatric surgery ward for evaluation of GERD as a cause of chronic respiratory symptoms by 24 hours PH monitoring. Additionally, 10 patients with only one episode of pneumonia were evaluated as the control group. Chronic respiratory presentations include the following: chronic cough, recurrent pneumonia, asthma, and respiratory distress. RESULTS: 24 hour esophageal PH monitoring revealed GER in 22 (42.2%) patients as a cause of their chronic respiratory symptoms, while (30 (57.7%) children did not show any evidence of GER. GER was detected in 11 of 24 (45.7%) patients with chronic cough. Thirty three patients presented with recurrent pneumonia, 13 (39.9%) of whom had GER. In 8 patients with asthma, GER was found in 4 cases. None of the 6 patients with respiratory distress had GERD. CONCLUSION: The possibility of GERD was significantly higher in study group (children with chronic respiratory symptoms) compared to control group (p-value<0.01). All patients with chronic cough, recurrent pneumonia and asthma should be aggressively investigated for the possibility of GER. Documenting abnormal gastroesophageal reflux helps direct appropriate therapy before occurrence of major complications. PMID- 11929030 TI - The congenital long QT syndrome. AB - OBJECTIVE: The long QT syndrome (LQTS) is a disorder of the electrical system of the heart, due to dysfunction of the ion channels and involving the repolarisation process. The inherited form occurs when there is a mutation in one of the genes which encode the making of a channel. Prolongation of the QT interval renders the patient vulnerable to an arrythmia called torsade de pointes, resulting in syncope and sudden death. METHODS: Three children with the congenital long QT syndrome presented to the pediatric department, one of them also having a 2:1 atrio-ventricular block. The parents and siblings of these children were screened for the long QT syndrome with an electrocardiogram. 2D echocardiography was done to rule out structural abnormalities and audiometry for deafness. RESULTS: Four family members were identified on screening to have LQTS. Propranolol was started on all children with LQTS. The child with heart block also received a pacemaker. LQTS must be considered in all patients presenting with syncope especially if associated with deafness and/or a family history of sudden deaths in infancy or childhood. CONCLUSION: The corrected QT interval must be determined in all children with heart block since the two conditions are often associated. PMID- 11929031 TI - Psychological consequences of chronic physical illnesses in children and adolescents. AB - Chronic illnesses in children and adolescents have devastating influence on them and their families. The patients have to cope up with illness, medication and it's influence on their development. Consequently a large number of them have emotional disorders which influence the course and outcome of physical disorder. In recent few years, research has shifted its focus from merely figuring out the prevalence of the emotional disorders to understanding the adjustment to chronic illness under defined theoretical constructs. Apart from temperamental variation, family burden and functioning have been identified as factors operating in determining adjustment. Distress experienced by the family and disturbed family functioning directly influences the emotional outcome in physically disordered children. Comprehensive coordinated care services are required for integration of these patients into the community. PMID- 11929033 TI - Upper gastrointestinal bleeding: etiology and management. AB - Upper gastrointestinal bleeding is a potentially fatal condition at times due to loss of large volumes of blood. Common sources of upper gastrointestinal bleeding in children include mucosal lesions and variceal hemorrhage (most commonly extra hepatic portal venous obstruction) and, in intensive care settings infections and drugs are other etiological factors associated with bleeding. Massive upper GI bleeding is life threatening and requires immediate resuscitation measures in the form of protection of the airways, oxygen administration, immediate volume replacement with ringer lactate or normal saline, transfusion of whole blood or packed cells and also monitoring the adequacy of volume replacement by central venous lines and urine output. Upper GI endoscopy is an effective initial diagnostic modality to localize the site and cause of bleeding in almost 85-90% of patients. Antacids supplemented by H2- receptor antagonists, proton pump inhibitors and sucralfate are the mainstay in the treatment of bleeding from mucosal lesion. For variceal bleeds, emergency endoscopy is the treatment of choice after initial haemodynamic stabilization of patient. If facilities for endoscopic sclerotherapy (EST) are not available, pharmacotherapy which decreases the portal pressure is almost equally effective and should be resorted to. Shunt surgery is reserved for patients who do not respond to the above therapy. Beta blockers combined with sclerotherapy have been shown to be the most effective therapy in significantly reducing the risk of recurrent rebleeding from varices as well as the death rates, as compared to any other modality of treatment. Based on studies among adult patients, presence of shock, co-morbidities, underlying diagnosis, presence of stigmata of recent hemorrhage on endoscopy and rebleeding are independent risk factors for mortality due to upper GI bleeding. Rebleeding is more likely to occur if the patient has hematemesis, liver disease, coagulopathy, hypotension and or anemia. There is a great need for conducting therapeutic trials as well as identifying predictors of outcome of upper GI bleeding in children to develop evidence based management protocols. PMID- 11929032 TI - Management of chronic hepatitis B and C virus infections. AB - Hepatitis B and C virus (HBV and HCV) infections present an important health problem causing significant morbidity and mortality on a worldwide scale. The younger the subjects infected, the higher the risk predisposing to progression towards chronic infection. Treatment of chronic HBV and HCV infections is aimed at reducing hepatic inflammation and thus improving the symptoms, decreasing the likelihood of long-term sequelae such as hepatocellular carcinoma, and increasing the survival rate. Interferon accelerates the spontaneous course of chronic HBV infection in children with greater disease activity and lower levels of replication. There is limited information on the use of lamivudine and its long term benefit in children with chronic HBV infection. The response of combination therapy with IFN and ribavirin in children with chronic HCV infection is still under investigation. The long-term clinical and virological effects of various drugs used in chronic HBV and HCV infections on children remain to be evaluated. PMID- 11929034 TI - Ascites and encephalopathy in chronic liver disease. AB - Ascites and particularly encephalopathy in the setting of chronic liver disease are traditionally thought to be poor prognostic markers of end stage liver disease. Diagnostic and therapeutic approaches to these complications are based on experience with adult patients, however, differences in the spectrum of aetiologies causing end stage liver disease together with the relative resistance of the pediatric patient to encephalopathy impact on the management of such patients. The following is a review of current thinking in the aetiology and management of ascites and encephalopathy in the setting of chronic liver disease in children. PMID- 11929035 TI - Liver transplantation in children. AB - The results of liver transplantation has improved significantly in the last decade with one year survival figures close to 90% for children with chronic liver disease. This can be attributed to improvement in surgical techniques, better postoperative care and newer immunosuppresive drugs. As a result of this, increasing number of children are referred for transplantation with no significant increase in the number of solid organ donors. The earliest transplants in children were performed using organs from size matched pediatric donors. However, as the pediatric donor numbers were limited, liver reduction techniques were developed to transplant small children before deterioration. Increasing experience with reduced livers led to the development of split liver, living donor and auxiliary liver transplantation. Better management of immunosuppressive drugs and newer agents such as Mycophenolate Mofetil have reduced the incidence of graft loss due to chronic rejection and long-term renal toxicity. The goal for the future will remain to be transplantation without the use of long-term immunosuppression. PMID- 11929037 TI - Blount's disease: a lesser known cause of bowlegs mandating early differentiation from physiological bowing. AB - We present a 16-month-old child, with progressively increasing bowing of legs, having a normal serum calcium, phosphorus and alkaline phosphatase level; lower limb radiographs revealed bilateral medial tibial metaphyseal beaking. Based on these findings, a diagnosis of Blount's disease (infantile tibia vara) was made and an orthotic management program was instituted for the child. PMID- 11929036 TI - Chemotherapy related fatal neurotoxicity during induction in acute lymphoblastic leukemia. AB - Neurotoxicity is a common complication during cancer chemotherapy. It is estimated that 3-10% of children with acute lymphoblastic leukemia (ALL) experience acute, transient neurotoxicity during induction chemotherapy. Fatal acute neurotoxicity is rarely encountered. Neurological evaluation of children with ALL at diagnosis and during treatment is of value in order to diagnose neurological complications early so that appropriate intervention can be adopted. This communication describes the profile of two children with unexpected, acute fatal neurologic toxicity during induction chemotherapy for ALL. PMID- 11929038 TI - Sjogren-Larsson syndrome. AB - Sjogren-Larsson Syndrome is a rare autosomal disorder which occurs with 100% penetrance and is classically characterized by ichthyosis, spasticity and mental handicap. This has rarely been described in people of Indian origin, case reports mainly being restricted to Caucasians. We have described a 6-year-old girl with classical features of this syndrome, the symptoms having started in infancy. The spasticity is mainly diplegic in nature. Skin changes of ichthyosis are generalized and more in the flexural areas. Mental retardation is severe. Management is supportive. PMID- 11929039 TI - Complications of varicella zoster. AB - Primary infection with varicella zoster is characterzed by a generalized vesicular rash usually without significant systemic illness. Encephalitis, pneumonitis, pancreatitis, nephritis, Reye and Guillan-Barre syndrome transvers myelitis, myocarditis have been reported before, but there is not any case having all these system to be involved during the same infection in a sequential manner ending up with multiorgan failure. We wanted to represent 21-month-old boy had a multiorgan failure due to varicella zoster infection. PMID- 11929040 TI - Renal tubular acidosis presenting with respiratory arrest. PMID- 11929041 TI - The effects of some porphyrinogenic drugs on the brain cholinergic system. AB - In central nervous system, acetylcholinesterase (AChE) and butyrylcholinesterase (BuChE) hydrolyse acetylcholine. Diminished cholinesterase activity is known to alter several mental and psychomotor functions. The symptoms of cholinergic crisis and those observed during acute attacks of acute intermittent porphyria are very similar. The aim of this study was to investigate if there could be a link between the action of some porphyrinogenic drugs on brain and the alteration of the cholinergic system. To this end, AChE and BuChE activities were assayed in whole and different brain areas. Muscarinic acetylcholine receptor (mAChR) levels were also measured. Results obtained indicate that the porphyrinogenic drugs tested affect central cholinergic transmission. Quantification of mAChR gave quite different levels depending on the xenobiotic. Veronal administration inhibited 50% BuChE activity in whole brain, cortex and hippocampus; concomitantly cortex mAChR was 30% reduced. Acute and chronic isoflurane anaesthesia diminished BuChE activity by 70-90% in whole brain instead cerebellum and hippocampus mAChR levels were only altered by chronic enflurane anaesthesia. Differential inhibition of cholinesterases in the brain regions and their consequent effects may be of importance to the knowledge of the mechanisms of neurotoxicity of porphyrinogenic drugs. PMID- 11929042 TI - Circular permutation of 5-aminolevulinate synthase as a tool to evaluate folding, structure and function. AB - 5-Aminolevulinate synthase, a pyridoxal 5'-phosphate-dependent enzyme, catalyzes the condensation of glycine with succinyl-coenzyme A to yield aminolevulinate, carbon dioxide and CoA. This reaction corresponds to the first and regulatory step of the mammalian heme biosynthetic pathway. Mutations in the erythroid aminolevulinate synthase gene are associated with X-linked sideroblastic anemia, an erythropoietic disorder characterized by the presence of hypochromic microcytic erythrocytes in peripheral blood and ring sideroblasts in bone marrow. In the past five years, transient kinetic studies in conjunction with three dimensional structure models and engineered variants of aminolevulinate synthase have been instrumental in understanding the individual steps of the catalytic mechanism of aminolevulinate synthase. The mechanism of folding, assembly of the two subunits into a functional, dimeric holoenzyme has been recently explored in this laboratory using circular permutation of aminolevulinate synthase. PMID- 11929043 TI - Porphyrin profiles in blood and urine as a biomarker for exposure to various arsenic species. AB - A sensitive method using HPLC with fluorescence detection has been established for the measurement of porphyrins in biological materials. The assay recoveries were 88.0+/-1.8% for protoporphyrin IX in the blood, and ranged from 98.3+/-2.7% to 111.1+/-7.4% for various porphyrins in the urine. This method was employed to investigate the altered porphyrin profiles in rats after a single dose of various arsenicals including soluble sodium arsenate and sodium arsenite, and the relatively insoluble calcium arsenite, calcium arsenate and arsenic-contaminated soils at dose rates of 5 mg/kg or 0.5 mg/kg body weight. Porphyrin concentrations increased within 2448 hr after the arsenic treatment in blood and urine. Protoporphyrin IX is the predominant porphyrin in the blood. In rats administered 5 mg As(III)/kg body weight, protoporphyrin IX concentration elevated to 123% of the control values in rats, 24 hr after the treatment. Higher increases were recorded in the urinary protoporphyrin IX (253% at 24 hr; 397% on day 2), uroporphyrin (121% at 24 hr; 208% on day 2) and coproporphyrin III (391% at 24 hr; 304% on day 2), while there was no significant increase (109% on day 3) observed in the urinary coproporphyrin I excretion. In rats administered 5 mg As(V)/kg, urinary excretion of protoporphyrin LX, uroporphyrin, coproporphyrin III and coproporphyrin I elevated to the maximum levels by 48 hr with the corresponding percentage values compared to the control being 177%, 158%, 224% and 143%, respectively. In rats dosed with 5 mg As(III)/kg, the increases (expressed as % of the control values) of protoporphyrin IX in the blood were in the order: sodium arsenite (144%) > sodium arsenate (125%) > calcium arsenite (123%) > calcium arsenate. In contrast, there was no significant increase of protoporphyrin IX, when the six arsenic-contaminated cattle dip soils and nine copper chrome arsenate (CCA-contaminated) soils were administered to the rats. Probable explanations are discussed. PMID- 11929044 TI - Levels of uroporphyrinogen decarboxylase (URO-D) in erythrocytes of Italian porphyria cutanea tarda patients. AB - Porphyria cutanea tarda (PCT) is a human metabolic disorder due to the acquired or genetic impairment of uroporphyrinogen decarboxylase (URO-D) activity, the fifth enzyme of the heme biosynthetic pathway. A classification of inherited and non-inherited forms is based on the enzyme activity levels in red blood cells (RBC). Clinical manifestations of PCT are often precipitated by triggering factors such as alcohol, drug abuse, estrogens, virus infections, hepatotoxic chemicals and hepatic siderosis. We measured URO-D activity in RBC from a large sample of Italian PCT patients in order to define the enzyme activity distribution and to attempt a correlation among activity, risk factors and clinical outcome. Three classes of patients with low, normal and over-normal URO D activity were defined according to control values. Low URO-D levels were present in 25.8% of patients, suggesting the familial form of PCT (type II). In this group, the outcome of PCT seems to be less influenced by risk factors. Patients with over-normal URO-D activity in RBC deserve further investigation. PMID- 11929045 TI - Hemochromatosis (HFE) and transferrin receptor-1 (TFRC1) genes in sporadic porphyria cutanea tarda (sPCT). AB - Porphyria cutanea tarda (PCT), a disorder characterized by a photosensitive dermatosis and hepatic siderosis, is caused by a decreased activity of the hepatic enzyme uroporphyrinogen decarboxylase (UROD). Two forms of PCT have been described: a familial one (fPCT) with an inherited decrease of UROD activity in all tissues and a sporadic one (sPCT) with a decreased UROD activity restricted to the liver. Iron overload and acquired factors including hepatic viral infections, alcohol, drugs contribute to the expression of PCT. In 65 French sPCT patients and 108 controls we have evaluated the respective role of iron and HCV status, the hemochromatosis (HFE) gene mutations frequencies (H63D. S65C, C282Y), and in a case control study we searched for an association between sPCT and the human transferrin receptor-1 (TFRC1) gene whose product is thought to be in functional association with the HFE protein: three single nucleotide polymorphisms (SNPs) previously characterized and 2 novel ones were studied. The iron-related parameters and transaminases were higher in sPCT patients than those of non-porphyric controls. Of the sPCT patients studied, 28% were HCV positive. In the HFE gene, 17% of sPCT patients carried C282Y mutation compared to 4% in controls, no significant differences were found with H63D and S65C variants. Compound heterozygous genotypes, C282Y/H63D or C282Y/S65C, were not significantly different in sPCT and control groups. Independently from HFE gene mutations, an association was found between the IVS4+198 T allele in the TFRC1 gene and sPCT patients. Analysis of HFE genotypes indicated that C282Y (but not H63D nor S65C) is a susceptibility factor for the development of sPCT in West European continental patients. However, analysis of TFRC1 genotypes suggest that sPCT should be considered as a multifactorial disorder in which other intracellular iron metabolism genes could be involved. PMID- 11929046 TI - Autoimmunity and HCV infection in porphyria cutanea tarda: a controlled study. AB - Autoimmunity and high rates of autoantibodies have been implicated in the pathogenesis of porphyria cutanea tarda. These abnormalities could be in part virus-induced, since porphyria cutanea tarda in most geographical regions is highly associated with hepatitis C virus infection. We analyzed the link of autoantibodies, autoimmune hepatitis and systemic lupus erythematosus in 111 patients with porphyria cutanea tarda and sex- and age-matched controls (mean age 58+/-13 years) in Germany, a region with a low prevalence of hepatitis C virus infection. Patients with porphyria cutanea tarda displayed lower rates of anti nuclear antibodies (16/111, 14% vs 28/111, 25%, p<0,05) and of antibodies against smooth muscle (25/111, 23% vs 48/111, 43%, p<0,01), than controls. The percentage of patients with porphyria cutanea tarda with positive anti-HCV was low but significantly higher than in our controls (9/111, 8% vs 0/111, 0%, respectively), (p<0,05). Two patients with porphyria cutanea tarda (2/111, 2%) fulfilled the criteria for systemic lupus erythematosus and not one of 65 patients was found to have clinical autoimmune hepatitis. In the first controlled study of a large cohort of patients with porphyria cutanea tarda no increased prevalence of selected autoantibodies and autoimmune hepatitis was found. However, a higher prevalence of HCV infection and systemic lupus erythematosus in patients with porphyria cutanea tarda was confirmed. PMID- 11929047 TI - Molecular, immunological, enzymatic and biochemical studies of coproporphyrinogen oxidase deficiency in a family with hereditary coproporphyria. AB - A 27-year-old woman who had recurrent pain in renal bed since 1998 with increasing character, was stationary admitted. The patient showed dark urine, complained of hair loss and took since 1994 a hormonal oral contraceptive. No photosensitivity was observed. Determinations of urinary porphyrin metabolites in 1998 revealed a porphyria cutanea tarda like excretion pattern with elevations of uro- (1767 nmol/24 hr, normal <29 nmol/24 hr) and heptacarboxyporphyrin (568 nmol/24 hr; normal <4 nmol/24 hr). Follow-up studies in feces showed the characteristics of a hereditary coproporphyria with dominance of coproporphyrin isomer III (total= 1470 nmol/g, isomer III= 93%), (normal: <37 nmol/g, isomer III = 25-35%). The excretion of porphyrin precursors (delta-aminolevulinic acid and porphobilinogen) was increased by taking an ethinylestradiol-cyproteronacetate preparation, but acute and/or chronic manifestations were not observed. Coproporphyrinogen oxidase activity was decreased to 35% in the patient (normal=138+/-21 pkat/g protein; x+/-s), whereas the activity of red cell uroporphyrinogen decarboxylase was normal. Her mother and both sisters could be verified as heterozygous gene carriers of hereditary coproporphyria by their urinary and fecal excretion parameters and because of reduced coproporphyrinogen oxidase activity up to 50%. The father was normal with respect to his genotype. Molecular analysis revealed a hitherto unknown mutation with the transversion of a cytosine to thymine at nucleotide position 854 in exon 4 of the coproporphyrinogen oxidase gene. The gene defect was confirmed by DGGE in the mother and her three daughters. The investigation of the immunological nature of the defective coproporphyrinogen oxidase gene from the whole family revealed decreased concentrations of coproporphyrinogen oxidase protein in the patient, her mother and her two sisters. PMID- 11929048 TI - Multiple mechanisms for hereditary sideroblastic anemia. AB - Hereditary sideroblastic anemia (HSA) is a heterogeneous group of inherited anemic disorders which is characterized by the presence of ringed sideroblasts in the bone marrow, microcytic hypochromic anemia and typically its X-linked inheritance in patients. It has been shown that a deficiency of the erythroid specific delta-aminolevulinate synthase (ALAS-E) activity is responsible for pyridoxine-responsive HSA in many patients, however, the pathogenesis of other types of HSA remains still unknown. In this article, recent evidence suggesting multiple causes for HSA is summarized and discussed. PMID- 11929049 TI - Characterization of the mouse protoporphyrinogen oxidase gene. AB - The murine protoporphyrinogen oxidase gene has been isolated, characterized and localized. The gene spans 4.2 kb, is comprised of 13 exons and 12 introns, and is located on chromosome 1 in band 1 H2. Analysis of 1.2 kb of the 5' upstream region revealed a promoter which is not GC rich and lacks any TATA boxes or initiator elements in the vicinity of the transcription start site. A variety of putative transcriptional element binding sequences were identified and gel shift assays support the presence of two GATA-1 sites near -760 bp as well as AP-1, AP 2, and Sp1 sites in the -1200 bp 5' flanking region. Luciferase reporter constructs transiently expressed in erythroid cell lines demonstrated erythroid specific expression with the -1160 bp, but not with the -746 bp or -198 bp constructs. Expression in nonerythroid cells occurred maximally with -1160 bp, but was significant with -746 bp and absent with -198 bp. Expression of both housekeeping and erythroid-specific fusions in the transient expression systems was greatly decreased in the -5000 bp constructs suggesting the presence of repressor elements in the -1160 to -5000 bp region. PMID- 11929050 TI - A mouse model for South African (R59W) variegate porphyria: construction and initial characterization. AB - Variegate porphyria is inherited as an autosomal dominant disease with variable penetrance. It is characterized clinically by photocutaneous sensitivity and acute neurovisceral attacks, and biochemically by abnormal porphyrin excretion in the urine and feces. While the world-wide incidence of variegate porphyria is relatively low, in South Africa it is one of the most common genetic diseases in humans. Due to the large number of patients with variegate porphyria in South Africa, and the fact that variegate porphyria is representative of both the so called "acute" and the "photocutaneous" porphyrias, it would be valuable to have an animal model in which to study the disease. In this study we have produced a mouse model of "South African" variegate porphyria with the R59W mutation in C57/BL6 mice via targeted gene replacement. Hepatic protoporphyrinogen oxidase activity was reduced by approximately 50% in mice heterozygous for the mutation. Urine and fecal samples from these mice, in the absence of exogenous inducers of hepatic haem synthesis, contain elevated concentrations of porphyrins and porphyrin precursors in a pattern similar to that found in human variegate porphyric subjects. Bypassing the rate-limiting step in haem biosynthesis by feeding 5-aminolevulinic acid to these mice, results in an accentuated porphyrin excretory pattern characteristic of the variegate porphyric phenotype and urinary porphobilinogen is increased significantly. This initial characterization of these mice suggest that they are a good model for variegate porphyria at the biochemical level. PMID- 11929051 TI - Functional studies of mutations in the human protoporphyrinogen oxidase gene in variegate porphyria. AB - The autosomal dominant disorder, variegate porphyria (VP), results from mutations in the protoporphyrinogen oxidase (PPOX) gene. We have investigated the effects of 22 disease-associated missense mutations in this gene on enzyme activity. Mutants were generated in the expression plasmid pHPPOX by site-directed mutagenesis. They were screened for PPOX activity by complementation of the Escherischia coli strain SAS38X which lacks PPOX activity. Ten mutants (G40E, L85P, G232R, de1281H, V282D, L295P, V335G, S350P, L444P, G453V) had no detectable PPOX activity. PPOX activity of the remaining 12 mutants (L15F, R38P, L73P, V84G, D143V, R152C, L154P, V158M, R168H, A172V, V290L, G453R) ranged from less than 1% to 9.2% of wild-type activity. Our findings show that all 22 mutations substantially impair or abolish PPOX activity in a prokaryotic expression system and add to the evidence that they cause VP. PMID- 11929052 TI - Ferrochelatase gene mutations in erythropoietic protoporphyria: focus on liver disease. AB - A deficiency of ferrochelatase (FECH) activity underlies the excess accumulation of protoporphyrin that occurs in erythropoietic protoporphyria (EPP). In some patients, protoporphyrin accumulation causes liver damage that necessitates liver transplantation. The purpose of this study was to determine if specific mutations in the FECH gene are present in patients who develop liver disease. FECH cDNA and all 11 exons and their flanking intron regions in the FECH gene were amplified and sequenced by specific polymerase chain reactions. Gene mutations were determined in 34 individuals from 24 families: 14 had liver disease, 10 necessitating liver transplantation. All individuals were heterozygous for mutations that altered the coding region of FECH mRNA. The mutations in patients with liver disease were heterogenous, but usually caused a major structural alterations in the FECH protein, most commonly as a result of exon skipping in FECH mRNA. However, the mutations could not account for the severe phenotype by themselves, since the same mutations were found in asymptomatic family members of patients with liver disease and in patients from families in which liver disease was not present. Other genetic factors, and possibly acquired factors, also must be critical to the development of this severe phenotype in EPP. PMID- 11929053 TI - A genotype-phenotype correlation between null-allele mutations in the ferrochelatase gene and liver complication in patients with erythropoietic protoporphyria. AB - Erythropoietic protoporphyria (EPP), an inborn error of heme metabolism, causes in the majority of the patients only a symptom of photosensitivity. However, around 2% of the EPP sufferers develop liver complication in the form of liver cirrhosis and progressive liver failure. Mutations in the human ferrochelatase (FECH) gene causing EPP are highly heterogeneous and mostly family-specific. Actually, 62 FECH mutations have been published, 48 of them are "null allele" mutations inducing the formation of a truncated protein. The remaining 14 are missense mutations. In contrast to the null allele mutations, the latter lead to substitution of a single amino acid residue in the protein molecule and generate an enzyme that, although functionally impaired, is in its full length. In order to study the association between "null allele" mutation and liver complication, we combined our data with those in the literature. A total of 112 EPP patients were counted among 93 EPP families with a known FECH mutation. All 18 EPP patients who had severe liver complication carried a "null allele" mutation. In contrast, none of the 20 patients who carried a missense mutation had developed liver complication till the time of study (Fisher's exact test, p<0.05). High protoporphyrin blood concentration are considered to be a sign of an increased risk of liver disease. No correlation of protoporphyrin blood level with the type of mutation, was found, if patients with overt liver disease were excluded from the sample. Furthermore, no significant association of the liver complication with the location of the mutation within the FECH gene was found (Fisher exact test p = 0.46). These available data indicate a significant genotype-phenotype correlation between "null allele" mutation and protoporphyrin related liver disease in EPP. Although the risk for a EPP patient with a missense mutation to develop liver disease cannot be totally eliminated based on these data, it is comparably low. PMID- 11929054 TI - Late-onset porphyrias: what are they? AB - Porphyrias are inherited disorders of heme biosynthesis. ALA dehydratase porphyria (ADP) and congenital erythropoietic porphyria (CEP) are autosomal recessive porphyrias, and are typically expressed at birth or in childhood. However, a few cases of late-onset recessive porphyrias have been reported. Recently we encountered a late-onset ADP patient who developed symptoms of acute porphyria when he was 63 years old. This was accompanied by polycythemia vera. It was concluded that he developed the porphyria because an abnormal ALAD allele was clonally expanded by polycythemia vera. Upon reviewing the literature, a few cases of late-onset CEP were found to be also associated with hematologic abnormalities suggestive of myelodysplastic syndrome (MDS), another clonal disorder. These findings suggest that these late-onset porphyrias may be heterozygous for their gene defects, but clinical expression may be elicited if there is a loss of heterozygosity, either by a clonal expansion of the porphyric allele or by a loss of function mutation in the other allele. PMID- 11929055 TI - Morbidity on youth expeditions to developing countries. AB - OBJECTIVE: To document the frequency of illness and injury on youth expeditions and to compare this information with published data concerning other travelers. METHODS: During July and August 1996, 945 young people aged 15 to 20 years traveled to the developing world with a single expedition company, World Challenge Expeditions Ltd. All groups were supervised by adult leaders. Leaders recorded all episodes of illness or injury occurring on the expedition on medical cards. We report a retrospective analysis of these medical cards. RESULTS: There were no deaths or emergency repatriations for the entire group of 65 expeditions. A complete set of medical cards was received from 47 of 65 expeditions (72%). Data were available for 644 individuals (309 male and 335 female). Local medical advice was sought on 17 occasions (1% of all episodes); 2 expeditioners required admission to a hospital. Four hundred eighty individuals (75%) experienced 1646 recorded health problems during their expedition. The most common problems were headache (18% of individuals), diarrhea (33%), nausea and/or vomiting (20%), and respiratory tract symptoms (14%). CONCLUSIONS: This study suggests supervised travel to the developing world is safe for young people. The incidence of gastrointestinal and respiratory symptoms was similar to that in travelers of other age groups, and trauma or serious injury was much less common in this group than is reported in other travelers. Headache was more common than in older travelers and indicates that preexpedition briefing should focus on the assessment and potential significance of this symptom in the younger age group, particularly at high altitude (above 2500 m). PMID- 11929056 TI - Effectiveness of a repellent containing DEET and EBAAP for preventing tick bites. AB - OBJECTIVE: Topical repellents can provide effective personal protection from tick borne diseases by preventing the attachment of ticks. The goal of this study was to assess the effectiveness of a commercially available repellent spray containing both N,N-diethyl-3-methylbenzamide, previously known as N,N-diethyl-m toluamide (DEET), and ethyl-butylacetylaminopropionate (EBAAP) against tick bites in a population at risk in Switzerland under real-life conditions. METHODS: The effectiveness of an insect repellent spray containing both DEET and EBAAP was evaluated in a randomized, double-blind, placebo-controlled field study. The study, requiring simple application of the repellent to exposed skin, was carried out on 276 forestry workers and orienteers under everyday conditions in Switzerland from May to September 1999. We measured total effectiveness of the repellent by the following formula: percentage effectiveness = 100 x (T(P) - T(R))/T(P), where T(P) and T(R) were the average number of ticks per hour spent in wooded areas for the repellent and placebo groups, respectively. RESULTS: The average number of tick bites per hour of exposure to wooded areas differed significantly between the placebo (n = 138) and repellent (n = 138) groups, 0.17 vs 0.10 (P < .05). Total repellent effectiveness against tick attachment was 41.1% (95% CI, 2.5-79.6). On the arms, an effectiveness of 66% (95% CI, 17.3 114.7) was observed. No significant difference in the average number of unattached ticks could be found. CONCLUSIONS: This study found that an easily applied repellent is moderately effective in reducing the risk of tick bites. PMID- 11929057 TI - Evaluation of diagnostic criteria and incidence of acute mountain sickness in preverbal children. AB - OBJECTIVE: The Children's Lake Louise Score (CLLS) established the diagnostic criteria for acute mountain sickness (AMS) in preverbal children. Prospective application of the CLLS and interobserver agreement for the score had not been evaluated in a controlled trial. A study of children and their parents was used to evaluate the CLLS and determine the incidence of AMS in preverbal children. METHODS: A prospective, controlled trial. Children > or =3 months and < or =36 months old and their parents living below 1645 m were studied over 7 separate days. The CLLS, measured daily by the mother as well as by the father on days 5 and 6, is the sum of scores for fussiness (FS), eating (E), playfulness (P), and sleep (S). Children were studied on days 1 and 2 at home, on day 3 after travel without altitude gain to a hotel, on day 4 at home, on days 5 and 6 at a hotel at 3109 m, and on day 7 at home. Using our previous criteria, AMS was diagnosed if the CLLS was > or =7 with both the FS > or =4 and the E + P + S > or =3. Agreement between mothers' and fathers' CLLS values was measured with the kappa statistic (K). Adults were also evaluated for AMS by the CLLS on days 5 and 6. RESULTS: Thirty-seven children (mean age +/- SD = 16.5+/-10.5 months; 21 girls) participated, and AMS occurred in 7 of them (19%; 95% CI, 8.35%). Among 33 adults, 8 (24%; 95% CI, 9.39%) had AMS. Although the agreement of the parents on the CLLS components was poor, the agreement on the classification of AMS between mother and father was excellent (K = .67; P < .001), with both parents' scores exceeding the CLLS threshold for AMS. CONCLUSIONS: In this prospective trial, parents demonstrated excellent interobserver agreement for independent use of the CLLS to detect AMS. Consistent with our previous retrospective study, the incidence of AMS at moderate altitude in preverbal children (19%) was similar to that in adults (24%). PMID- 11929058 TI - Mechanisms of injury in competitive off-road bicycling. AB - OBJECTIVE: To describe the mechanisms of injury from crashes during competitive off-road bicycling (mountain biking) and to examine the type, location, and severity of the resulting injuries. METHODS: We examined and interviewed all bicyclists injured while competing at 7 off-road bicycling events. Information regarding the direction of fall and the presence of a collision or mechanical failure was obtained, and the injury patterns were compared. RESULTS: There were 97 injured riders, with a mean age of 28.3 years. Most victims were male (74%), and all cyclists wore helmets. Most injuries were minor and involved the extremities (70.5%). Injuries sustained from falling forward over the handlebars occurred more often than from falling to the side (65% vs 25%), tended to lead to injuries that were more severe (mean injury severity score [ISS] = 3.4 vs 1.7, P < .05), and produced more head and neck injuries (56% vs 8%, P < .05). Falls to the side generally led to a lower extremity injury (88% vs 57%, P < .05). Riders who were involved in collisions had injuries that were similar in severity and location to those of riders who had no collision. CONCLUSIONS: These findings suggest that off-road bicyclists whose mechanism of injury involves falling forward over the handlebars are at risk for more severe injury, especially to the head and neck. PMID- 11929059 TI - Acute mountain sickness in tourists with children at Lake Chungara (4400 m) in northern Chile. AB - OBJECTIVE: To evaluate the presence of acute mountain sickness (AMS) and cardiorespiratory responses in adult and pediatric tourists exposed to high altitude (Putre and Chungara). METHODS: The study was performed during a tourist expedition to Lake Chungara (4400 m) with an overnight stay in the town of Putre (3500 m). The study group included 15 adults (21-44 years), 10 teenagers (13-18 years), and 6 children (6-48 months). We evaluated AMS symptoms in adults with the Lake Louise questionnaire and in children with the modified Children's Lake Louise Score for preverbal subjects. Cardiorespiratory measurements included heart rate and oxygen saturation on arrival at Putre and Chungara. Values were expressed as mean +/- SD; significance was tested by analysis of variance, followed by the Newman-Keuls test (P < .05). RESULTS: Significantly lower hemoglobin saturation was found among the children in Putre (74.8% +/- 5.2%) and Chungara (65.3%+/-1.3%) than among the teenagers (82.2%+/-3.2% and 78.6% 2.5%) and adults (83.9%+/-3.3% and 79.7%+/-5.5%) at a similar altitude. A higher incidence of AMS was observed in children (100%) in Putre than in teenagers (50%) and adults (27%). CONCLUSIONS: Our results indicate that children are extremely sensitive to hypoxia, as expressed by symptoms of AMS and significant desaturation. Our findings add to the available information regarding the problems encountered when ascending to high altitude with children and support the importance of close monitoring of young children during ascent to high altitude. PMID- 11929060 TI - A comparison of mountain rescue casualty bags in a cold, windy environment. AB - OBJECTIVE: To help the Mountain Rescue Association of Scotland find the best protective mountain rescue casualty bag in cold and windy conditions. The study investigated how 3 different casualty bags (labeled Bag 1, Bag 2, and Bag 3) performed in a cold (-10 degrees C, dry bulb), windy (wind speed 3.0 m x s(-1)) environment using physiological and subjective responses of the participating subjects. METHODS: Eleven male subjects, aged 23.4+/-4 years, percentage body fat 15.5+/-2 (mean +/- SD). Each participated in a total of 3 tests (1 for each bag). The tests were scheduled to last 60 minutes. Core and skin temperatures (skin values were measured on the arm, chest, thigh, and calf, and a mean skin temperature was calculated) were measured during the tests. Heart rate, oxygen consumption (Vo2), and subjective cold perception ratings were also recorded at regular intervals throughout the test duration. All variables except for Vo2 and cold discomfort were adjusted for baseline. RESULTS: There was a significant difference in the mean response between the bags for the following variables: arm, chest, thigh, calf, mean temperature, and cold discomfort. CONCLUSIONS: All 3 bags showed limited ability to protect the subjects in cold, windy conditions. However, the study shows that Bag 2 offered the least protection against the imposed environment. It is difficult to differentiate between the other 2 bags, because Bag 1 performed better than Bag 3 for arm, calf, and mean temperatures, while Bag 3 outperformed Bag 1 for chest and thigh temperatures and cold discomfort scores. PMID- 11929061 TI - Acute leptospirosis in a triathlete. AB - We report the case of a 38-year-old male patient with acute leptospirosis. The most probable cause of infection was repeated and prolonged exposure to contaminated river water (Neckar and Enz rivers) while preparing for participation in long-distance triathlon (swimming, biking, and running) competitions. PMID- 11929062 TI - Skin lesions caused by stink bugs (Insecta: Heteroptera: Pentatomidae): first report of dermatological injuries in humans. PMID- 11929063 TI - Anthropological and thermoregulatory changes induced by a survival sojourn in a tropical climate. AB - OBJECTIVE: A survival sojourn in a tropical climate exposes subjects to 2 main constraints: food restriction and tropical climatic conditions. We hypothesized that such a sojourn could modify anthropological characteristics and thermoregulatory responses to heat and cold on return. METHODS: Eight European male subjects were submitted to a sweating test (ST) for 90 minutes (dry bulb temperature [Tdb] = 47 degrees C, relative humidity (RH) = 10%, and wind speed = 0.8 m x s(-1)) and to a whole-body cold-air test (Tdb = 1 degrees C, RH = 40%, and wind speed = 0.8 m x s(-1)) for 120 minutes in thermoclimatic chambers both before and after a 4-week survival sojourn in French Guyana. RESULTS: The survival sojourn resulted in a decrease in lean body mass (P < .05) without any significant change in body fat content. Heat thermoregulatory changes studied during the ST were characterized both by a lower mean skin temperature (Tsk) (P < .05) and a higher sweat rate measured after the sojourn than before it (m(sw); P < .05). Cold thermoregulatory changes were characterized by a higher T(sk) value (P < .05) and a decreased onset for continuous shivering without any significant change in internal temperature or metabolic heat production (M). CONCLUSIONS: This study showed that this type of sojourn modifies not only biometrical characteristics of the subjects but also the thermoregulatory responses, inducing an adaptation of the thermoregulatory system to heat and an increase in the sensitivity of the thermoregulatory system to cold. This finding could have practical implications on return after such a sojourn. PMID- 11929064 TI - Hysterical conversion disorder in a young girl at moderate altitude. PMID- 11929066 TI - The "Silver Hut" expedition--a commentary 40 years later. PMID- 11929065 TI - The ascent of Mount Everest following radial keratotomy. PMID- 11929067 TI - Mountaineering and climbing techniques in the curriculum of mountain medicine education programs: a survey of the European Courses for Mountain Medicine. AB - Mountain medicine education programs have been described and discussed from several perspectives. A Union Internationale des Associations Alpinistes Internationale Kommission fur Alpines Rettungswesen-International Society for Mountain Medicine (UIAA-IKAR-ISMM) label was defined and subsequently awarded to courses that comply with certain minimal requirements. These courses can award an international UIAA-IKAR-ISMM diploma to their successful candidates. The need for mountaineering training was listed in those minimal requirements, but no exact recommendations were given. A complete analysis of existing mountain medicine education programs in Europe is done, providing information about the practical mountaineering and climbing techniques taught in the different courses and, if applicable, the practical requirements and examinations to obtain the above mentioned diploma. The analysis clearly shows the widely varying requirements for proficiency in mountaineering and climbing in European mountain medicine courses. PMID- 11929068 TI - Writing good test questions. PMID- 11929069 TI - The removal of NH3-N from primary treated wastewater in subsurface reed beds using different substrates. AB - Subsurface flow experimental reed beds, were designed and built based on a combination of two design methodologies, that of the WRc and Severn Trent Water plc (3) and that of the USA, EPA (17). Four different growing media were used with a combination of top soil, gravel, river sand and mature sewage sludge compost, aiming to determine the best substrate for ammonia removal. Eight units were constructed, two for each material. One bed for each pair was planted with Typha latifolia plants commonly known as cattails. Primary treated domestic wastewater, was continuously fed in to the bed for more than six months. The best results were achieved by the gravel reed beds with an almost constant removal rate of NH3-N above 80%. There was no significance difference on the performance of planted and unplanted reed beds. PMID- 11929070 TI - Filamentous fungi in Indah Water Konsortium (IWK) sewage treatment plant for biological treatment of domestic wastewater sludge. AB - A study was carried out to isolate and identify filamentous fungi for the treatment of domestic wastewater sludge by enhancing biodegradability, settleability and dewaterability of treated sludge using liquid state bioconversion process. A total of 70 strains of filamentous fungi were isolated from three different sources (wastewater, sewage sludge and leachate) of IWK's (Indah Water Konsortium) sewage treatment plant, Malaysia. The isolated strains were purified by conventional techniques and identified by microscopic examination. The strains isolated belonged to the genera of Penicillium, Aspergillus, Trichoderma, Spicaria and Hyaloflorae The distribution of observed isolated fungi were 41% in sewage sludge followed by 39% in wastewater and 20% in leachate. The predominant fungus was Penicillium (39 strains). The second and third most common isolates were Aspergillus (14 strains) and Trichoderma (12 strains). The other isolates were Spicaria (3 strains) and Hyaloflorae (2 strains). Three strains (WWZP1003, LZP3001, LZP3005) of Penicillium (P. corylophilum, P. waksmanii, and P. citrinum respectively), 2 strains (WWZA1006 and SS2017) of Aspergillus (A. terrues and A. flavus respectively) and one strain (SSZT2008) of Trichoderma (T. harzianum) were tentatively identified up to species level and finally verified by CABI Bioscience Identification Services, UK. PMID- 11929071 TI - Improved method for concentration of Giardia, Cryptosporidium, and poliovirus from water. AB - Methods for the concentration of enteric viruses and the protozoan parasites, Giardia and Cryptosporidium, from drinking water currently require the use of two different types of filters. Electropositive or electronegative microporous filters (0.2-0.45 microm nominal porosity) are used for the collection of enteroviruses, while polypropylene spun-fiber filters (1 microm porosity) and small pleated cartridge filters are used for the collection of protozoan parasites from water. Since the filter mechanically traps the protozoa by size exclusion, a microporous filter with an appropriately small nominal porosity could possibly be used for co-collection of both protozoa and enteroviruses. This study compared the concentration efficiencies of a polypropylene fiber cartridge (DPPPY) filter and two different microporous filters (Filterite and IMDS) with poliovirus (type 1), with respect to their ability to concentrate Giardia and Cryptosporidium from water. Giardia cysts and Cryptosporidium oocysts were added to 4001 of either tap water or tertiary treated wastewater and passed through the test filter. The protozoa were eluted from the polypropylene filter by hand washing in a detergent solution. Viruses and protozoa were eluted from the microporous filter by two consecutive back-washes with a 1.5% beef extract, 0.1% Tween 80 solution. The eluent was then centrifuged to remove the parasites and the supernatant assayed for viruses. The overall efficiency was greater for the Filterite filter (40.4% for Giardia; 36.6% for Cryptosporidium) when compared to the spun fiber filter (10.1% for Giardia; 16.0% for Cryptosporidium). The Filterite filters were easier and faster to process than the polypropylene spun fiber filters. There was no significant difference in the recovery of protozoa from 1MDS and DPPPY filters. Co-collection of viruses and protozoan parasites from water onto the same filter is possible and can reduce the time and cost of routine water monitoring. PMID- 11929072 TI - Speciation and bioavailability of EDTA complexed copper in the microenvironment of fish gills. AB - Carp were exposed to a synthetic solution with varied concentrations of Cu-EDTA and constant levels of other copper species and the amount of copper accumulated in the fish gills was measured. The distributions of copper species in both the bulk solution and the fish gill microenvironment were modeled using MINTEQA2. Under experimental conditions without EDTA, Cu(OH)2 was the most dominant species in the bulk solution while Cu-mucus dominated the gill microenvironment. When Cu EDTA was added to the solution, copper accumulation in the fish gills increased significantly, suggesting a change in copper speciation. The amount of copper accumulated in the fish gills correlated positively to the concentration of Cu mucus, rather than to free copper. PMID- 11929073 TI - Mercury in mushrooms and soil of the Tarnobrzeska Plain, south-eastern Poland. AB - Mercury was quantified in the fruiting bodies of 15 species of higher mushrooms and underlying soil substrate collected from the Tarnobrzeska Plain in south eastern part of Poland in 1995. In total, 405 samples each of caps, stalks or whole fruiting bodies and 221 samples of soil (0 10cm layer) were examined. The area under investigation can be considered generally as unpolluted with mercury since the mean concentrations in underlying soil substrate of 14 mushroom species were between 21 +/- 9 and 58 +/- 31 ng/g dry wt. (total range between 4.5-240). Somewhat elevated concentrations, of 630 +/- 80 ng/g dry wt., were noted for underlying soil substrate of Livid Entoloma (Entoloma sinuatum). Among the mushroom species examined, King Bolete (Boletus edulis), Livid Entoloma, and Red capped Scaber Stalk (Leccinum rufum) contained the greatest mercury concentrations both in the caps (between 1,800 +/- 600 and 3,500 +/- 1,100 ng/g dry wt.) and stalks (between 900 +/- 400 and 2,300 +/- 1,000 ng/g dry wt.). Because of the relatively great bioconcentration factors (BCF) of mercury, which averaged between 73 +/- 44 and 220 +/- 270 in the caps, and between 37 +/- 28 and 170 +/- 240 in the stalks, King Bolete, Bay Bolete (Xerocomus badius) and Red capped Scaber Stalk can be considered as good mercury accumulators. Nevertheless, apart from the Common Earth Ball (Scleroderma citrinum), that can be considered as an excluder of mercury due to the BCF value of less than 1, other species examined were characterised by BCFs greater than 1. The caps were usually characterised with greater mercury content than the stalks (Hg(Cap)/Hg(stalk) quotients > 1) and the reverse was noted only for Bay Bolete. Due to a statistically significant relationship between mercury content of the caps/stalks and underlying soil substrate (p<0.001) in Filamentous Pax (Paxilus filamentosus), and in the stalks of Red-capped Scaber (p < 0.05) and Brick Tops (Hypholoma sublateritium) (caps; p < 0.05), these species can be suggested to exhibit bioindication potential for soil mercury contamination. PMID- 11929074 TI - Butyltins in sediments and three-spined stickleback (Gasterosteus aculleatus) from the marinas of the Gulf of Gdansk, Baltic Sea. AB - Fifty four sediment samples and three pooled samples of whole three-spined stickleback (Gasterosteus aculleatus) collected from four marinas located along the south-western coast of the Gulf of Gdansk, Baltic Sea, Poland, were examined for butyltins (BTs) i.e., tributyltin (TBT), dibutyltin (DBT) and monobutyltin (MBT), to understand the degree of pollution and distribution pattern. Total butyltin concentrations in sediments from the marina of the city of Gdynia ranged between 130 and 20,000 ng Sn/g dry wt. with a mean value of 2,900 ng Sn/g. In sediments collected from the marinas located in the area of the Brave Vistula River Channel BT concentrations ranged between 10 to 15,000 ng Sn/g drywt. with a mean values between 500 and 5,500 ng Sn/g, and the innermost sites of each of four marinas were the most polluted. The mean BT concentrations in three-spined stickleback were between 1,500 and 3,100 ng Sn/g wetwt. and the values followed mean concentrations noted in sediments from the corresponding marina. The pleasure yachts are identified as the main source of BT pollution of sediment and fish. Tributyltin was a major form of BTs present in sediments (between 24 and 43%) and in three-spined stickleback (between 54 and 70%), suggesting a very recent use of marine paints containing TBT in pleasure yachts. Evidently at the Polish coast of the Baltic Sea TBT was continuously used in 1997 in marine paints for boats less then 25 m long. PMID- 11929075 TI - Kinetic studies on UV-photodegradation of some chlorophenols using TiO2 catalyst. AB - A number of chlorophenols, namely 2-chlorophenol, 2,4,-dichlorophenol and 2,4,6 trichlorophenol, were decomposed in aqueous solution by using TiO2 as photoactivated catalyst under UV radiation emitted by a 125W medium pressure Hg lamp in an immersion well-type quartz photoreactor. The organic-bound chlorine was converted into the environmentally harmless inorganic chloride. For catalyst doses between 0.1 and 0.5gl(-1) the reaction mechanisms are elucidated. The corresponding rate constants were obtained by periodically measuring the remaining chlorophenol, and converted chloride in solution. A theoretical model for the degradation pathway is proposed expressing the rate as a linear function of the concentrations of chlorophenol and catalyst. Aside from the model calculated values, the pseudo-first order rate constants for a rough approximation of chlorophenols degradation as well as the kinetic parameters of Langmuir-Hinshelwood type decomposition are compared. The photodegradation rate of chlorophenols followed the order: C13 x Ph > Cl2 x Ph > Cl x Ph. PMID- 11929076 TI - Kinetics of adsorption and uptake of Cu2+ by Chlorella vulgaris: influence of pH, temperature, culture age, and cations. AB - Adsorption and uptake of Cu2+ by Chlorella vulgaris were distinguished by extracting the surface-bound Cu2+ with EDTA. The uptake of Cu2+ followed Michaelis Menten kinetics. The maximum rate of Cu2+ uptake (0.362fmolcell(-1) h( 1)) was obtained at pH 6.0. The rate of Cu2+ uptake was greater for cultures in the exponential phase of growth, and increased with a rise in temperature from 6 to 25 degrees C, thus pointing towards an active mechanism. The maximum number of Cu2+ binding sites was 3.245 fmol cell(-1) at pH 4.5. Adsorption of Cu2+ was strongly pH-dependent thereby indicating that the number and nature of metal binding sites on the cell surface change with changing chemistry of the solution. Unlike uptake, the adsorption remained unaffected by small changes in temperature. Older cultures displayed a higher Cu2+ adsorption capacity than the exponentially growing ones thus suggesting generation of new and/or additional Cu2+ binding sites on older cells of C. vulgaris. By pH titration, the cation exchange capacity of Chlorella, measured in terms of H+/ Na+ exchange, was about 17 fmol cell(-1) at pH 10.5. Negligible cation exchange capacity at and below pH 5.0 indicated that ion exchange was not the sole mechanism of Cu2+ adsorption by Chlorella. The uptake and adsorption of Cu2+ were inhibited by 100 microM of various cations including other heavy metal ions. The general concept that cations competitively inhibit accumulation of metals in living organisms does not hold for C. vulgaris. Non-competitive, uncompetitive and mixed inhibition of Cu2+ uptake and adsorption by various cations were more common than competitive inhibition. PMID- 11929077 TI - Decolorization of methyl red by a mixed culture of Bacillus sp. and Pseudomonas stutzeri. AB - The decolorization of methyl red directed toward rapid reduction of azo group by a mixed culture of Bacillus sp. and Pseudomonas stutzeri was investigated. The improvement of the decolorization of methyl red was obtained by a mixed culture (a mixed culture 2) with particular proportion of two bacteria (Bacillus sp. and Pseudomonas stutzeri). The mixed culture 2 was grown in a LB medium with methyl red. The improvement of the decolorization of methyl red was obtained only by the growing mixed culture and not obtained by the mixture of the resting cells. PMID- 11929078 TI - Effects of sibutramine in non-dieting obese women. AB - The aim of this study was to evaluate the effects of sibutramine on plasma leptin levels, body weight and glucose metabolism in non-dieting women. Fourteen healthy, non-diabetic, obese women were studied before treatment, after 1 week of placebo administration, and after a 2-week course of sibutramine (10 mg/day). At each of these stages, we assessed body composition, measured the levels of plasma leptin, C-peptide and various biochemical parameters, and also recorded plasma insulin and glucose levels during oral glucose tolerance tests. After 1 week of placebo treatment, there were no significant changes in any of the parameters. However, two weeks of 10 mg/day sibutramine dropped plasma leptin levels from a mean (+/-SE) of 48.84+/-4.54 to 42.84+/-4.74 ng/ml (p<0.04), reduced BMI from 39.36+/-2.01 to 38.57+/-1.93 kg/m2 (p<0.002), and decreased insulin resistance (IR, as measured using the homeostasis model assessment of insulin resistance) from 5.59+/-0.85 to 3.66+/-0.43 (p<0.02). There was no correlation between the reduction in leptin concentration and the decrease in BMI, fat mass, percent body fat, IR, C-peptide, or the area under curve for glucose or insulin. There was also no correlation between the decrease in leptin levels and the increases that occurred in the insulin sensitivity index or the hepatic sensitivity index. The results showed that treatment with 10 mg/day sibutramine significantly reduces BMI, IR and leptin levels in non-dieting obese women. PMID- 11929079 TI - Levothyroxine therapy and serum free thyroxine and free triiodothyronine concentrations. AB - Although the normal thyroid gland secretes both levothyroxine (L-T4) and levotriiodothyronine (L-T3), normalization of serum TSH with L-T4-replacement therapy alone in hypothyroidism is generally believed to result in a normal serum L-T3 and to reflect a euthyroid state. However several recent studies suggest that this may not be the case. Accordingly, the relationship between serum free L T4 and free L-T3 was examined in 20 normal individuals (group A) and in 53 patients with chronic autoimmune thyroiditis, 18 with normal TSH on no L-T4 replacement (group B), and 35 with normal TSH on L-T4-replacement therapy for hypothyroidism (group C). Data were analyzed by applying a one-way analysis of variance with correction for multiple comparisons. Serum TSH values were very similar among the 3 groups. In groups A and B, mean serum free T4 and free T3 were very similar. In group C, the mean free T4 (16+/-2 pmol/l) was significantly higher than the values in groups A (14+/-1) and B (14+/-2) (p<0.001) and the mean free T3 lower (4.0+/-0.5 pmol/l vs 4.2+/-0.5, NS and 4.4+/-0.5, p<0.02). Consequently, the mean molar ratio of free T4 to free T3 was significantly higher in group C than the ratios in groups A and B (p<0.0001), despite very similar TSH values. These findings indicate that in hypothyroid patients L-T4-replacement, that is sufficient to maintain a normal serum TSH, is accompanied by a serum free T4 that is higher than that in untreated euthyroid patients or normal individuals and may not result in an appropriately normal serum free T3 concentration. PMID- 11929080 TI - Prognostic value of serial serum thyroglobulin determinations after total thyroidectomy for differentiated thyroid cancer. AB - Serial weekly serum samples (for 3 weeks) were obtained from 42 patients with differentiated thyroid cancer (DTC, papillary no.=35, follicular no.=6, Hurthle cell no.=1) for serum thyroid hormone, TSH and TG before and after total thyroidectomy. Serum specimens were also obtained one month after radioiodine (131I) therapy followed by suppressive dose of L-thyroxine (L-T4, 2.5 microg/kg). The patients were subdivided into four groups: group I: the DTC was confined to a single solid nodule (no.=1 2); group II: thyroid malignancy invaded local cervical structures but there were no lymph node metastases (no.=8); group III: DTC with lymph node metastases (no.=6); and group IV: DTC with distant metastases (no.=16). In all group I patients serum TG remained undetectable in spite of elevated serum TSH levels at the 3rd week post-surgery (PS). Only one of group II patients had a detectable serum TG value of 5.2 ng/ml (3rd week PS). By contrast, 37.5% of group III patients had detectable serum TG levels, ranging from 3.4 to 16.8 ng/ml (3rd week PS). Lymph node metastases were detected in 5 of these patients by whole body scan (WBS) and removed surgically in 3. As expected, group IV patients had elevated serum TG values ranging 33.0-958.0 ng/ml and distant metastases were confirmed in all of them by WBS. From the calculations through univariate logistic regression comparing TG concentrations at the 3rd week PS from groups I and II vs groups III and IV, we obtained a cut-off value of 2.3 ng/ml with the following efficacy features: sensitivity=74.5%; specificity=95%; positive predictive value=92.3%; negative predictive value=65.5%; and accuracy=73.8%. After 131I and L-T4 suppressive therapy, only 5 out of 36 patients of groups I, II and III had detectable serum TG levels (3.1-7.0 ng/ml) whereas serum TG was detectable in all group IV patients (ranging 2.5-8.6 ng/ml). We concluded that serum TG concentrations above 2.3 ng/ml at the 3rd week PS could be suggestive of lymph node or distant metastases in patients with DTC. Patients with serum TG above this limit could be considered at risk for metastatic disease and higher doses of diagnostic iodine-131 (131I) may be indicated for actinic ablation. PMID- 11929081 TI - Activity of GH/IGF-1 axis in burn patients: comparison with normal subjects and patients with GH deficiency. AB - The aim of this study was to clarify the activity of GH/IGF-1 axis as well as the variations of nutritional parameters following a thermal injury in man. To this goal, in 22 patients with burn [BURN, age (mean+/-SE): 46.5+/-3.4 yr, BMI: 25.0+/ 0.8 kg/m2, % burn surface area: 26.0+/-3.0%, ROI score: 0.22+/-0.1] we evaluated IGF-1, IGF binding protein (IGFBP-3), GH, GH binding protein (GHBP), pre-albumin (pre-A), albumin (A) and transferrin (TRA) levels on days 1, 3, 7, 14 and 28 after intensive care unit (ICU) admission. IGF-1, IGFBP-3, GH and GHBP levels were also assayed basally in 29 normal subjects (Ns) (Ns, age: 47.5+/-2.8 yr, BMI: 22.0+/-1.4 kg/m2) and in 34 panhypopituitary patients with severe GH deficiency (GHD, age: 42.7+/-2.5 yr, BMI: 25.6+/-0.8 kg/m2). On ICU day 1, IGF-1 and IGFBP-3 in BURN were higher than those in GHD (p<0.05 for both, respectively) and lower than those in Ns (p<0.05) while GH levels in BURN did not differ from those in Ns and higher than GHD (p<0.01). In BURN, IGF-I and IGFBP-3 levels showed a progressive decline (p<0.05) with nadir on day 14, when they overlapped those in GHD, and then an increase on day 28, though persisting lower than in Ns, while GH levels did not vary during ICU stay. IGF-I levels were associated neither to burn extension nor to ROI score. On ICU day 1 pre-A, A and TRA levels were similar to those in Ns, but underwent a progressive decrease with nadir on day 7 (p<0.001) for pre-A and TRA, and later, on day 14 (p<0.05) for A; pre-A and TRA but not A showed a rebound increase (p<0.01) on day 14, though persistingly lower than in Ns. In conclusion, our present data firstly show the time course variation of IGF-I levels in burn patients as function of nutritional and hormonal variables. It has to be emphasized that in the most critical phase after burn injuries, IGF-1 levels are as low as in hypopituitary patients with severe GHD. The physiological basis which leads to the impairment of this endogenous anabolic drive in this phase is, however, not clear yet. PMID- 11929083 TI - Effects of high doses of corticosteroids on bone metabolism. AB - The effects of a chronic treatment with corticosteroids on bone are well known, but few data are available regarding the acute effect of these drugs on bone turnover. This study was aimed at evaluating the effects of high doses of corticosteroids administered for a short period on bone metabolism. We assessed 23 subjects (15 women and 8 men) suffering from multiple sclerosis and treated with methylprednisolone (1 g i.v. for 10 days) followed by oral prednisone for 9 days; patients affected by diseases involving bone or treated during the previous 6 months with drugs influencing bone metabolism were excluded. We observed a significant decrease of ALP and bone glia protein (BGP), in these subjects, and a significant sudden increase of urinary calcium/creatinine and urinary cross-laps after 3 days of treatment. All of these parameters, except urinary calcium/creatinine, returned to basal levels after 30 days from the beginning of treatment (11 days after the interruption of corticosteroids administration). Serum phosphorus showed a significant decrease after 3 days of treatment, but returned to basal levels after 10 days. These data suggest that high doses of corticosteroids administered for a short period are able to induce an increase of bone resorption and a decrease of bone formation; moreover, bone turnover returns to basal levels when the treatment is stopped. PMID- 11929082 TI - Lymphocyte subset pattern in acromegaly. AB - Immune function in acromegalic patients has been poorly investigated. The aim of this study was to evaluate the main surface antigen clusters of circulating lymphocytes in acromegaly. One hundred patients with active acromegaly (55 women and 45 men, aged 20-70 yr) and 200 healthy subjects sex- and age-matched with the patients (110 women and 90 men, aged 20-70 yr) were enrolled in this study. All patients and controls were born and live in Southern Italy. No patient had received octreotide, bromocriptine or corticosteroids for at least 3 months before entering the study. The analysis of lymphocyte subset pattern was performed by flow cytometry and fluorescein isothiocyanate or phycoerythrin directly conjugated monoclonal antibodies specific for the cell surface antigen clusters (CD) representing T-cell population as a whole (CD3), T helpers (CD4), T suppressors (CD8), natural killer cells (CD16) and B-cell population as a whole (CD19). Acromegalics had significantly increased levels of CD3 (67.1+/-7.2 vs 64.3+/-8.8%; p=0.03) and CD4 (37.8+/-3.5 vs 36.4+/-4.3%; p=0.004) and decreased levels of CD8 (31.4+/-3.3 vs 33.7+/-8.2%; p<0.01) and CD19 (12.1+/-3.1 vs 15.2+/ 5.1; p=0.01) without age-difference. The results of the current study demonstrate an increase in T-cell activity together with a decrease in B-cell activity in a very large series of patients with active acromegaly. These data further support the existence of abnormalities of the immune system in patients with chronic GH/IGF-1 excess. PMID- 11929084 TI - Lower limb alactic anaerobic power output assessed with different techniques in morbid obesity. AB - Short-term alactic anaerobic performance in jumping (5 consecutive jumps with maximal effort), sprint running (8 m) and stair climbing (modified Margaria test) were measured in 75 obese subjects (BMI: 40.3+/-5.0 kg/m2) and in 36 lean control subjects (BMI: 22.4+/-3.2 kg/m2) of the same age and gender distribution. The results show that obese subjects attained a significantly lower specific (per unit body mass) power output both in jumping (W(spec,j); p<0.001) and stair climbing (W(spec,s); p<0.001) and run at a significantly lower average velocity (v; p<0.001) during sprinting. In spite of the different motor skillfulness required to accomplish the jumping and climbing tests, W(spec,s) (and hence the vertical velocity in climbing, v(v)) was closely correlated with W(spec,j) (R2=0.427, p<0.001). In jumping, although the average force during the positive work phase was significantly higher in obese subjects (p<0.001), no difference between the 2 groups was detected in absolute power. In stair climbing the absolute power output of obese resulted significantly higher (18%) than that of lean controls (p<0.001). In sprint running, the lower average horizontal velocity attained by obese subjects also entailed a different locomotion pattern with shorter step length (L(s); p<0.001), lower frequency (p<0.001) and longer foot contact time with ground (T(c,r); p<0.001). W(spec,j) seems to be a determinant of the poorer motor performance of obese, being significantly correlated with: I) the vertical displacement of the centre of gravity (R2=0.853, p<0.001) in jumping; II) with v(v) in stair climbing; and III) with T(c,r) (R2=0.492, p<0.001), L(s) (R2=0.266, p<0.001) and v (R2=0.454, p<0.001) in sprinting. The results suggest that obese individuals, although partially hampered in kinetic movements, largely rely on their effective specific power output to perform complex anaerobic tasks, and they suffer from the disproportionate excess of inert mass of fat. Furthermore, in view of the sedentary style of life and the consequent degree of muscle de-conditioning accompanying this condition, it may prove useful to implement rehabilitation programs for obesity with effective power training protocols. PMID- 11929085 TI - Evaluation of circulating levels and renal clearance of natural amino acids in patients with Cushing's disease. AB - Although the hypercortisolism-induced impairment of protein homeostasis is object of several studies, a detailed evaluation of the complete amino acid profile of patients with Cushing's syndrome (CS) has never been performed. The aim of the current open transversal controlled study was to evaluate serum and urinary concentrations as well as renal clearance of the complete series of natural amino acids and their relationship with glucose tolerance in patients with Cushing's disease (CD). Twenty patients with CD (10 active and 10 cured) and 20 sex- and age-matched healthy controls entered the study. Measurement of serum and urinary levels of the complete series of natural amino acids was performed in all patients analyzed by cationic exchange high performance liquid cromatography (HPLC) after 2 weeks of a standardized protein intake regimen. The renal clearance (renal excretion rate) of each amino acid was calculated on the basis of the serum and urinary concentrations of creatinine and the specific amino acid. Fasting glucose and insulin levels, glucose and insulin response to standard glucose load, insulinogenic and homeostasis model insulin resistance (Homa-R) indexes were also evaluated and correlated to the circulating levels and renal clearances of each amino acid. Significantly higher serum (p<0.01) and urinary (p<0.05) levels of alanine and cystine, lower serum and higher urinary levels of leucine, isoleucine and valine (p<0.05) and higher renal excretion rates of leucine, isoleucine and valine (p<0.01) were found in patients with active CD than in patients cured from the disease and in controls. No difference was found between cured patients and controls. Creatinine clearance was similar in active and cured patients and in controls. In patients with active CD, urinary cortisol levels were significantly correlated to urinary cystine levels (r=0.85; p<0.01) and renal excretion rate of leucine (r=-0.76; p<0.05), isoleucine (r= 0.76; p<0.05) and valine (r=-0.66; p<0.05). Fasting blood glucose levels were significantly correlated to serum alanine levels (r=0.70; p<0.05). Although Homa R was significantly correlated to BMI in active patients (r=0.74 p<0.05), it was not correlated to amino acid levels. In conclusion, the results of the current study demonstrate that patients with CD have significant changes in serum and urinary concentration of several amino acids and changes in renal clearance of some specific amino acids. Normalization of cortisol levels restored the amino acid profile. PMID- 11929086 TI - Does smoking increase relapse rates in Graves' disease? AB - We wondered whether the relapse rate of Graves' hyperthyroidism was increased amongst patients who smoked. To this end, a retrospective analysis of clinical and laboratory data of consecutive patients with Graves' disease was carried out. All patients were treated with thionamide anti-thyroid drugs (ATD) for at least one year and remission was defined as continued and biochemical euthyroidism, at least 6 months after discontinuing ATD. The study comprised 221 subjects with Graves' disease from a hospital-based population over 9 years. We took the following variables into account when assessing contribution to disease relapse: Goiter, Thyroid Associated Ophthalmopathy (TAO), and Time to euthyroidism after starting ATD and interaction between smoking and sex. Smoking had a marginally significant (p=0.081) deleterious effect on the likelihood of remission after ATD treatment for Graves' disease. The effect of smoking was, however, highly significant in males and indeed the deleterious effect on remissions may be restricted to males (odds ratio, 11.1; 95% confidence interval, 1.25 to 98.5). The presence of goiter (odds ratio, 3.8; 95% confidence interval, 2.05 to 7.1) and TAO (odds ratio, 1.8; 95% confidence interval, 0.993 to 3.18) forecasted lower chances of achieving remission. The shorter the time a patient became euthyroid after starting ATD the more likely his disease was to remit. We conclude that cigarette smoking increases the likelihood of Graves' disease recurrence in males treated with anti-thyroid drugs. Thus, smoking appears to be an important risk factor in the pathogenesis and outcome of Graves' disease patients at least in subsets of patients. PMID- 11929087 TI - Traumatic hypopituitarism due to maternal uterine leiomyomas. AB - Hypopituitarism has been associated with different types of head trauma including traumatic delivery. We report a case of hypopituitarism in a boy with a history of induced traumatic labor associated with maternal uterine leiomyomas. He also had head and face deformations that were apparently caused by spatial restriction due to the enlarging leiomyomas while the patient was growing in utero. Trauma to the pituitary stalk could have occurred by cerebral entrapment and the pressures of labor. Although hypopituitarism has been associated with traumatic delivery and breech delivery, there are no reported cases related to uterine leiomyomas. PMID- 11929089 TI - Subacute thyroiditis in an immunosuppressed patient. AB - Subacute thyroiditis is a well-recognized cause of transient thyrotoxicosis, resulting from a destruction injury to the thyroid. The pathogenesis of this condition is not completely understood and there is debate regarding the extent of the contribution of autoimmunity and external agents, such as infections, to this process. We present the first reported case of subacute thyroiditis in a patient who had been on chronic lithium therapy as well as long-term immunosuppression, with cyclosporin and prednisolone, following an allogeneic bone marrow transplant. We speculate that this case suggests a minimal role of autoimmunity in the development of subacute thyroiditis. PMID- 11929088 TI - Very delayed hyponatremia after surgery and radiotherapy for a pituitary macroadenoma. AB - Severe hyponatremia (118 mmol/l) with natriuresis, consistent with cerebral salt wasting syndrome (CSWS), occurred 38 days after transsphenoidal surgery in a 59 year-old woman affected by a pituitary non-functioning macroadenoma. From the 35th day after surgery, she showed progressive polyuria, hypotension and hyponatremia associated with natriuresis, decreased plasma and increased urinary osmolality. The clinical examination revealed signs of dehydration and gradual decline in the level of consciousness. The anterior pituitary function was normal due to appropriate replacement of thyroid and adrenal axis. The patient was treated with saline administration until normal natremia and water balance were restored and neurological symptoms had completely disappeared. This case focuses on the unusually prolonged time of development of post-surgery hyponatremia, despite delayed symptomatic hyponatremia being reported to commonly occur 7 days after transsphenoidal surgery. Therefore, we would advise not to limit the periodic follow-up of the hydroelectrolytic balance to the first two weeks after surgery, but to prolong it until after discharge from hospital. In fact, an early diagnosis is of great importance to prevent permanent neurological damage or death. Since CSWS and syndrome of inappropriate secretion of ADH, the two disorders alternatively imputed to generate post-surgical hyponatremia, are characterized by different pathogenic mechanisms and require opposing therapeutic approaches, the occurrence of extracellular volume dilution or of increased sodium renal loss should be carefully investigated. The evidences in favor of CSWS, the possible mechanisms behind the syndrome and diagnosis and management of patients with post-transsphenoidal surgery CSWS are discussed. PMID- 11929090 TI - Macroprolactinoma associated with Cushing's disease, successfully treated with cabergoline. AB - Multiple pituitary hormone hypersecretions have already been described, but the co-occurrence of PRL and ACTH excess is very rare. To our knowledge, medical treatment with cabergoline only, avoiding pituitary surgery and radiotherapy in this type of tumor has never been reported before. This case report deals with a 31-yr-old man affected with a macroprolactinoma associated with a florid clinical image of Cushing's disease. Normalization of the prolactin levels and the disappearance of clinical and biochemical features of Cushing's disease were obtained after administration of medical treatment only. PMID- 11929091 TI - Iopanoic acid rapidly controls type I amiodarone-induced thyrotoxicosis prior to thyroidectomy. AB - Amiodarone-induced thyrotoxicosis (AIT) may develop either in apparently normal thyroid glands (Type II AIT) or in the presence of sub-clinical thyroid abnormalities (either autonomous goiter or latent Graves' disease; Type I AIT). Mixed forms also occur. While Type I AIT is due to iodine-induced excess thyroid hormone synthesis, Type II AIT is a form of amiodarone (possibly iodine) -induced destructive thyroiditis. Type I AIT is usually treated by combined thionamide and potassium perchlorate therapy, but may be resistant to therapy. On the other hand, Type II AIT often responds favorably to glucocorticoids and may not require further therapy once euthyroidism has been restored. Not infrequently, however, AIT (especially Type I) is resistant to conventional treatment, and several weeks or months may elapse before euthyroidism is restored. Thyroidectomy has been carried out in Type I AIT patients, but thyroid surgery in thyrotoxic patients, especially those with underlying cardiac problems, carries a high surgical risk. In this study we describe 3 patients with Type I AIT, who were successfully treated with a short course of iopanoic acid (IOP), an oral cholecystographic agent, which is rich in iodine and is a potent inhibitor of 5'-deiodinase, resulting in a marked decrease in the peripheral tissue conversion of T4 to T3, in preparation for thyroid surgery. Euthyroidism was rapidly restored in 7-12 days, allowing a subsequent safe and uneventful thyroidectomy in all cases. These patients were then treated with L-T4 for their hypothyroidism and amiodarone was safely re-instituted. We suggest that IOP is the drug of choice in the rapid restoration of euthyroidism prior to definitive thyroidectomy in patients with drug resistant Type I AIT. PMID- 11929092 TI - Cushing's syndrome in children and adolescents: current diagnostic and therapeutic strategies. PMID- 11929093 TI - Complications of inferior petrosal sinus sampling for the etiological diagnosis of Cushing's syndrome. PMID- 11929094 TI - Interference causes false high calcitonin levels with a commercial assay. PMID- 11929095 TI - Bather. Fernando Botero--Medellin 1932. PMID- 11929096 TI - Interactions between GABAergic and aminoacidergic pathways in the control of gonadotropin and GH secretion in pre-pubertal female rats. AB - Present experiments were carried out in 23-day-old female rats to analyze the interaction between excitatory amino acids (EAAs) and gamma-aminobutyric acid (GABA) in the control of gonadotropin and GH secretion. For this purpose, serum concentrations of LH, FSH and GH were measured after injection of different agonists of EAA receptor subtypes [N-methyl-D-aspartate (NMDA); kainic acid (KA), (+/-)-alpha-amino-3-hydroxy-5-methylisoxazole-4-propionic acid (AMPA)], antagonists of GABA receptors (bicuculline, phaclofen) or the combined administration of both types of drugs. The results obtained indicated that: 1) GABA has a minor physiological role in the control of LH and GH secretion, since neither LH nor GH serum concentrations changed after administration of bicuculline (antagonist of GABA(A) receptors) or phaclofen (antagonist of GABA(B) receptors); 2) GABA has a sex-specific physiological role in the control of FSH secretion in female rats, in which FSH secretion increases after phaclofen administration; 3) GH secretion was enhanced after administration of NMDA, KA and AMPA, while LH increased only after activation of NMDA receptors; 4) the stimulatory effect of NMDA on LH secretion was counteracted by administration of phaclofen; and 5) bicuculline and phaclofen reduced the ability of NMDA and AMPA to stimulate GH secretion. In conclusion, present experiments evidenced a physiological role of GABA, mediated by GABA(B) receptors, in the control of FSH secretion and a cross-talk between excitatory and inhibitory amino acids in the control of anterior pituitary secretion. PMID- 11929097 TI - Genetic analyses and evaluation of peripheral parameters of thyroid hormone action for the differential diagnosis of RTH. A novel heterozygous missense mutation (M334T) discovered. AB - Resistance to thyroid hormone (RTH) is a rare disease characterized by goiter and elevated free thyroid hormone (TH) levels in the presence of detectable concentrations of TSH. Most RTH patients harbor mutations in the ligand binding domain (LBD) of thyroid hormone receptor beta (TRbeta) gene, without a clear correlation between genotype and phenotype. Clinical, biochemical and genetic analyses were performed in several members of one family, because the index case presented with elevated free TH, measurable TSH and no hyperthyroid manifestations, but with a pituitary lesion at MRI. High free TH levels and TSH concentrations in the normal range were found also in 4 relatives. The presence of euthyroidism in all patients together with peripheral parameters of TH action in the normal range led to the diagnosis of generalized RTH (GRTH). In the five affected members, the genetic analysis revealed a novel heterozygous missense mutation at codon 334 (M334T). A different mutation at codon 334 was previously described in association with selective pituitary resistance to thyroid hormone (PRTH). Therefore, we confirm that substitutions at Methionine 334 are critical for the structural integrity of TRbeta LBD. The association of different phenotypes with substitutions affecting the same codon is another contribution confirming that RTH phenotype does not generally depend upon the site of the mutation in the LBD of TRbeta1. PMID- 11929098 TI - Binary morphological shape-based interpolation applied to 3-D tooth reconstruction. AB - In this paper, we propose an interpolation algorithm using a mathematical morphology morphing approach. The aim of this algorithm is to reconstruct the n dimensional object from a group of (n - 1)-dimensional sets representing sections of that object. The morphing transformation modifies pairs of consecutive sets such that they approach in shape and size. The interpolated set is achieved when the two consecutive sets are made idempotent by the morphing transformation. We prove the convergence of the morphological morphing. The entire object is modeled by successively interpolating a certain number of intermediary sets between each two consecutive given sets. We apply the interpolation algorithm for three dimensional tooth reconstruction. PMID- 11929099 TI - Topology correction in brain cortex segmentation using a multiscale, graph-based algorithm. AB - Reconstructing an accurate and topologically correct representation of the cortical surface of the brain is an important objective in various neuroscience applications. Most cortical surface reconstruction methods either ignore topology or correct it using manual editing or methods that lead to inaccurate reconstructions. Shattuck and Leahy recently reported a fully automatic method that yields a topologically correct representation with little distortion of the underlying segmentation. We provide an alternate approach that has several advantages over their approach, including the use of arbitrary digital connectivities, a flexible morphology-based multiscale approach, and the option of foreground-only or background-only correction. A detailed analysis of the method's performance on 15 magnetic resonance brain images is provided. PMID- 11929101 TI - Automatic detection of abnormalities in chest radiographs using local texture analysis. AB - A fully automatic method is presented to detect abnormalities in frontal chest radiographs which are aggregated into an overall abnormality score. The method is aimed at finding abnormal signs of a diffuse textural nature, such as they are encountered in mass chest screening against tuberculosis (TB). The scheme starts with automatic segmentation of the lung fields, using active shape models. The segmentation is used to subdivide the lung fields into overlapping regions of various sizes. Texture features are extracted from each region, using the moments of responses to a multiscale filter bank. Additional "difference features" are obtained by subtracting feature vectors from corresponding regions in the left and right lung fields. A separate training set is constructed for each region. All regions are classified by voting among the k nearest neighbors, with leave one-out. Next, the classification results of each region are combined, using a weighted multiplier in which regions with higher classification reliability weigh more heavily. This produces an abnormality score for each image. The method is evaluated on two databases. The first database was collected from a TB mass chest screening program, from which 147 images with textural abnormalities and 241 normal images were selected. Although this database contains many subtle abnormalities, the classification has a sensitivity of 0.86 at a specificity of 0.50 and an area under the receiver operating characteristic (ROC) curve of 0.820. The second database consist of 100 normal images and 100 abnormal images with interstitial disease. For this database, the results were a sensitivity of 0.97 at a specificity of 0.90 and an area under the ROC curve of 0.986. PMID- 11929100 TI - Computation of the mid-sagittal plane in 3-D brain images. AB - We present a new method to automatically compute, reorient, and recenter the mid sagittal plane in anatomical and functional three-dimensional (3-D) brain images. This iterative approach is composed of two steps. At first, given an initial guess of the mid-sagittal plane (generally, the central plane of the image grid), the computation of local similarity measures between the two sides of the head allows to identify homologous anatomical structures or functional areas, by way of a block matching procedure. The output is a set of point-to-point correspondences: the centers of homologous blocks. Subsequently, we define the mid-sagittal plane as the one best superposing the points on one side and their counterparts on the other side by reflective symmetry. Practically, the computation of the parameters characterizing the plane is performed by a least trimmed squares estimation. Then, the estimated plane is aligned with the center of the image grid, and the whole process is iterated until convergence. The robust estimation technique we use allows normal or abnormal asymmetrical structures or areas to be treated as outliers, and the plane to be mainly computed from the underlying gross symmetry of the brain. The algorithm is fast and accurate, even for strongly tilted heads, and even in presence of high acquisition noise and bias field, as shown on a large set of synthetic data. The algorithm has also been visually evaluated on a large set of real magnetic resonance (MR) images. We present a few results on isotropic as well as anisotropic anatomical (MR and computed tomography) and functional (single photon emission computed tomography and positron emission tomography) real images, for normal and pathological subjects. PMID- 11929102 TI - A multiple circular path convolution neural network system for detection of mammographic masses. AB - A multiple circular path convolution neural network (MCPCNN) architecture specifically designed for the analysis of tumor and tumor-like structures has been constructed. We first divided each suspected tumor area into sectors and computed the defined mass features for each sector independently. These sector features were used on the input layer and were coordinated by convolution kernels of different sizes that propagated signals to the second layer in the neural network system. The convolution kernels were trained, as required, by presenting the training cases to the neural network. In this study, randomly selected mammograms were processed by a dual morphological enhancement technique. Radiodense areas were isolated and were delineated using a region growing algorithm. The boundary of each region of interest was then divided into 36 sectors using 36 equi-angular dividers radiated from the center of the region. A total of 144 Breast Imaging-Reporting and Data System-based features (i.e., four features per sector for 36 sectors) were computed as input values for the evaluation of this newly invented neural network system. The overall performance was 0.78-0.80 for the areas (Az) under the receiver operating characteristic curves using the conventional feed-forward neural network in the detection of mammographic masses. The performance was markedly improved with Az values ranging from 0.84 to 0.89 using the MCPCNN. This paper does not intend to claim the best mass detection system. Instead it reports a potentially better neural network structure for analyzing a set of the mass features defined by an investigator. PMID- 11929103 TI - Edge-preserving tomographic reconstruction with nonlocal regularization. AB - Tomographic image reconstruction using statistical methods can provide more accurate system modeling, statistical models, and physical constraints than the conventional filtered backprojection (FBP) method. Because of the ill posedness of the reconstruction problem, a roughness penalty is often imposed on the solution to control noise. To avoid smoothing of edges, which are important image attributes, various edge-preserving regularization methods have been proposed. Most of these schemes rely on information from local neighborhoods to determine the presence of edges. In this paper, we propose a cost function that incorporates nonlocal boundary information into the regularization method. We use an alternating minimization algorithm with deterministic annealing to minimize the proposed cost function, jointly estimating region boundaries and object pixel values. We apply variational techniques implemented using level-sets methods to update the boundary estimates; then, using the most recent boundary estimate, we minimize a space-variant quadratic cost function to update the image estimate. For the positron emission tomography transmission reconstruction application, we compare the bias-variance tradeoff of this method with that of a "conventional" penalized-likelihood algorithm with local Huber roughness penalty. PMID- 11929104 TI - Volume image registration by cross-entropy optimization. AB - Cross-entropy (CE), an information-theoretic measure, quantifies the difference between two probability density functions. This measure is applied to volume image registration. When a good prior estimation of the joint distribution of the voxel values of two images in registration is available, the CE can be minimized to find an optimal registration. If such a prior estimation is not available, one seeks the registration which gives a joint distribution different from unlikely ones as much as possible, i.e., the CE is maximized to find an optimal registration. When the unlikely distribution is a uniform one, CE maximization reduces to joint entropy minimization; when the unlikely distribution is proportional to one of the marginal distributions, it reduces to conditional entropy minimization; when the unlikely distribution is the product of two marginal distributions, it degenerates to mutual-information maximization. These different CEs are added together and are used as criteria for image registration. The accuracy and robustness of this new approach are tested and compared using a likely joint distribution and various unlikely joint distributions and their combinations. PMID- 11929105 TI - Finite difference time domain (FDTD) analysis of optical pulse responses in biological tissues for spectroscopic diffused optical tomography. AB - Finite difference time domain (FDTD) analysis has been successfully formulated for solving diffusion equation in biological tissues. Time-dependent diffusion equations are approximated by FDTD equations by assigning diffuse photon fluence rates and radiant flux defined in the diffusion equations to Yee meshes. At the boundary between scattering and no scattering material, FDTD equation including only fluence rate has been derived, which make it possible to calculate the fluence rate at the boundary. The formulation is useful to solve diffusion equations by iterative algebraic calculations in scattering media with inhomogeneous optical properties. The conditions to give stabilities for numerical solutions have been become clear in terms of scattering coefficients and mean cosine of scattering angles. Using the formulation, the reflectance of three-layered slabs containing a clear layer have been calculated. As a result, it has been found that absorption loss changes of the highly scattering medium beyond the clear layer are estimated from the time profiles of the reflectance. PMID- 11929106 TI - Initialization, noise, singularities, and scale in height ridge traversal for tubular object centerline extraction. AB - The extraction of the centerlines of tubular objects in two and three-dimensional images is a part of many clinical image analysis tasks. One common approach to tubular object centerline extraction is based on intensity ridge traversal. In this paper, we evaluate the effects of initialization, noise, and singularities on intensity ridge traversal and present multiscale heuristics and optimal-scale measures that minimize these effects. Monte Carlo experiments using simulated and clinical data are used to quantify how these "dynamic-scale" enhancements address clinical needs regarding speed, accuracy, and automation. In particular, we show that dynamic-scale ridge traversal is insensitive to its initial parameter settings, operates with little additional computational overhead, tracks centerlines with subvoxel accuracy, passes branch points, and handles significant image noise. We also illustrate the capabilities of the method for medical applications involving a variety of tubular structures in clinical data from different organs, patients, and imaging modalities. PMID- 11929107 TI - O(N3 log N) backprojection algorithm for the 3-D radon transform. AB - We present a novel backprojection algorithm for three-dimensional (3-D) radon transform data that requires O(N3 log2 N) operations for reconstruction of an N x N x N volume from O(N2) plane-integral projections. Our algorithm uses a hierarchical decomposition of the 3-D radon transform to recursively decompose the backprojection operation. Simulations are presented demonstrating reconstruction quality comparable to the standard filtered backprojection, which requires O(N5) computations under the same circumstances. PMID- 11929108 TI - Statistical image reconstruction for polyenergetic X-ray computed tomography. AB - This paper describes a statistical image reconstruction method for X-ray computed tomography (CT) that is based on a physical model that accounts for the polyenergetic X-ray source spectrum and the measurement nonlinearities caused by energy-dependent attenuation. We assume that the object consists of a given number of nonoverlapping materials, such as soft tissue and bone. The attenuation coefficient of each voxel is the product of its unknown density and a known energy-dependent mass attenuation coefficient. We formulate a penalized likelihood function for this polyenergetic model and develop an ordered-subsets iterative algorithm for estimating the unknown densities in each voxel. The algorithm monotonically decreases the cost function at each iteration when one subset is used. Applying this method to simulated X-ray CT measurements of objects containing both bone and soft tissue yields images with significantly reduced beam hardening artifacts. PMID- 11929109 TI - Foetal dosimetry-is the ICRP dosimetric system for humans now complete? PMID- 11929110 TI - Determination of 32P in urine for early estimation of the neutron exposure level for three victims of the JCO criticality accident. AB - In the criticality accident which occurred on 30 September 1999 at a uranium conversion facility in Tokai-mura, Japan, three workers were severely exposed to neutron and gamma-ray irradiation. Preliminary estimations of doses from blood properties and 24Na concentration in blood were 16-20, 6-10 and 1-4 gamma-ray gray-equivalent (gammaGyEq) respectively for the three workers. For apparent dose estimation, neutron-induced radionuclides in biological materials such as blood, hair and urine were measured. Accordingly, we detected 32p in urine samples. The concentration ratios of 32P in the urine for the three workers showed a similar tendency to those of 24Na in blood. This result indicated that the radioactivity of 32P in urine could be used to estimate the neutron exposure level. PMID- 11929111 TI - Over the border--the problems of uncontrolled radioactive materials crossing national borders. AB - Cross-border movement of radioactive materials and contaminated items, in particular metallurgical scrap, has become a problem of increasing importance. Radioactive sources out of regulatory control, now often called 'orphan sources', have frequently caused serious, even deadly, radiation exposures and widespread contamination. The United States Nuclear Regulatory Commission reported over 2,300 incidents of radioactive materials found in recycled metal scrap and more than 50 accidental smeltings of radioactive sources. A further potentially serious problem is illicit trafficking in nuclear and other radioactive materials. In 1995 the International Atomic Energy Agency (IAEA) started a programme to combat illicit trafficking in nuclear and other radioactive materials, which includes an international database on incidents of illicit trafficking, receiving reports from some 80 member states. For the period 1993 2000 the IAEA database includes 345 confirmed incidents. While from 1994-1996 the frequency declined significantly, this trend has been reversed since 1997, largely due to radioactive sources rather than nuclear material. This paper compares monitoring techniques for radioactive materials in scrap applied at steel plants and scrap yards with monitoring at borders, a completely different situation. It discusses the results of the 'Illicit Trafficking Radiation Detection Assessment Program', a large international pilot study, conducted in cooperation between the IAEA, the Austrian Government and the Austrian Research Centre Seibersdorf. The aim of this exercise was to derive realistic and internationally agreed requirements for border monitoring instrumentation. Finally the present extent of border monitoring installations is discussed. PMID- 11929112 TI - Radiation protection of people and the environment: developing a common approach. AB - The problem with the current ICRP system of radiation protection, particularly for regulators, is that it fails to differentiate between the application of justification and optimisation to people in the circumstances of medical care or as part of a workforce, compared with their application to members of the general public in an environmental setting; plus the fact that it also fails to address the issue of potential impacts on the rest of the environment in any meaningful way. But if these deficiencies are to be addressed, it will be essential to consider how protection of both people and the living environment can be achieved within a broad philosophical framework, using complementary approaches, based on the same underlying scientific knowledge. This paper briefly examines some of these issues, and offers some suggestions for developing a common, or even a combined, approach. It draws upon recent suggestions made by the ICRP itself with regard to radiological protection, plus current activities-on several fronts-to provide an explicit basis for environmental protection. PMID- 11929113 TI - Multiple parameter biodosimetry of exposed workers from the JCO criticality accident in Tokai-mura. PMID- 11929114 TI - Optimisation and common sense. AB - This note builds on recent articles about the development of new ICRP recommendations by supporting the use of common sense in optimisation; use of an additional criterion relating to technology-based principles is suggested to support utility- and equity-based criteria. This is taken forward by use of authoritative good practice safety precautions and a need to consider safety in an integrated manner. It is noted that use of common sense in ALARP or ALARA decisions is liable to rely on access to information and training. PMID- 11929115 TI - Caffeine ameliorates radiation-induced skin reactions in mice but does not influence tumour radiation response. AB - Intramuscular administration of caffeine at a dose of 80 mg kg(-1) body weight to the gastrocnemius muscles of Swiss mice 5 min prior to local irradiation (35 Gy) of the leg delayed the progression of radiation-induced skin reactions in such animals. While 90% epilation with reddening of the skin was noted in animals treated with radiation alone, animals pretreated with caffeine suffered only partial hair loss with slight reddening of the skin on the 16th and 20th days post-irradiation. Beyond the 28th day, damage scores in irradiated feet for both the groups were similar (score 3) and remained unchanged until the 32nd day and then decreased and disappeared completely in both treatment groups by the 40th day after irradiation. In addition, the effect of caffeine on the radiation response of a mouse fibrosarcoma was investigated. Results showed that intratumoral administration of caffeine at a dose of 80 mg kg(-1) body weight 5 min prior to local exposure of tumours to 10 Gy of 60Co gamma-rays did not influence the response of tumours to radiation. The present study thus showed that although caffeine ameliorated radiation-induced skin reactions in the mouse leg, it did not affect the tumour radiation response, indicating its potential application in cancer radiotherapy. PMID- 11929116 TI - Dose coefficients for the embryo and foetus following intakes of radionuclides by the mother. AB - Committee 2 of the International Commission on Radiological Protection (ICRP) has the responsibility for calculating radiation doses from intakes of radionuclides for all age groups in the population. Publication 88 of the ICRP, which has recently been published, describes the development of models used for calculating radiation doses to the embryo and foetus following intakes of radionuclides by the mother. It also gives radiation doses to the offspring for intakes of radionuclides by the mother either before or during pregnancy. The approaches used in the development of the biokinetic and dosimetric models are summarised here together with a comparison of the doses to the offspring with those to the reference adult. PMID- 11929117 TI - Getting the message across: is communicating risk to the public worth it? AB - The public and media often seem irrational in their conception of hazard and risk. Scare stories abound, and large sums of money may be spent to mitigate tiny risks. This article describes why it may be human not to be entirely logical about risk. The article also describes what makes risk professionals afraid of entering a public debate on risk. This information is then used to help define how we could improve communication with the public about risk. We discuss how the National Radiological Protection Board is tackling this now. PMID- 11929118 TI - Reply to 'comments on hereditary effects of radiation'. PMID- 11929120 TI - Healthy worker effect. PMID- 11929119 TI - Logic, collective dose and threshold dose. PMID- 11929123 TI - Depleted uranium: a study of its uses in the UK and disposal issues. PMID- 11929122 TI - The BSE inquiry. PMID- 11929124 TI - OX40: targeted immunotherapy--implications for tempering autoimmunity and enhancing vaccines. AB - OX40 (CD134), a membrane-bound member of the tumor-necrosis-factor-receptor superfamily, is expressed primarily on activated CD4+ T cells. Recently, several groups have reduced clinical signs of autoimmunity in animal models by blocking the OX40-OX40-ligand interaction or depleting OX40+ T cells. By contrast, engagement of OX40 in the setting of active immunization has potent adjuvant properties, leading to enhanced cytokine production and increased numbers of antigen-specific memory T cells. These potent adjuvant effects lead to an enhancement of anti-tumor responses. OX40 has several unique features that make it a clinically relevant target. They include: (1) T cells isolated from a site of inflammation that express OX40 are T cells that have been stimulated recentlythrough the T-cell receptor in vivo; (2) OX40 is only expressed on T cells found at the site of inflammation, therefore, targeting this receptor does not interfere with the peripheral T-cell repertoire; and (3) the biological function of OX40 is limited primarily to effector CD4+ T cells, which are a major source of cytokines to induce and maintain ongoing immune responses. PMID- 11929125 TI - Immunity and retroviral superantigens in humans. PMID- 11929126 TI - Complement in inflammatory tissue damage and disease. PMID- 11929127 TI - From A to Z on CpG. PMID- 11929128 TI - Dendritic cells and immune responses in the central nervous system. PMID- 11929129 TI - KIR expression shapes cytotoxic repertoires: a developmental program of survival. AB - We hypothesize a sequential program of expression of the leukocyte-receptor complex (LRC) in CD8+T cells, associated with cellular activation and the subsequent establishment of immune homeostasis through resistance to apoptosis. This program, which is consistent with the linear development of memory CD8+ T cells, represents an ordered expression of genes during differentiation, analogous to expression of the homeobox- or globin-gene clusters. Our model not only has implications for the development and maintenance of T-cell memory but also, relates to the formation of LRC repertoires in other cell types, particularly, the development of killer-cell Ig-like receptor (KIR) repertoires in natural-killer-cell precursors. PMID- 11929130 TI - The origin of CD95-gene mutations in B-cell lymphoma. AB - CD95 (Apo-1/Fas) is crucial for the negative selection of B cells within the germinal center (GC). Impairment of CD95-mediated apoptosis results in defective affinity maturation and the persistence of autoreactive B-cell clones. CD95 was defined recently as a tumor-suppressor gene and is silenced in many tumor entities. In contrast to other malignancies, in GC-derived B-cell lymphomas, inactivation of the CD95 gene is often a result of deleterious mutations. Such mutations occur also at a low frequency in normal GC, but not naive, B cells. We propose that CD95 mutations in B-cell lymphomas originate from the GC reaction and are introduced most probably as targeting errors of the somatic hypermutation machinery, which bears--besides its physiological role--an inherent risk of malignant transformation and the persistence of autoreactive B-cell specificities. PMID- 11929131 TI - Leukocyte Ig-like receptor complex (LRC) in mice and men. AB - Here, we compare the architecture of membrane receptors with extracellular Ig like domains located within the leukocyte Ig-like receptor complex (LRC) of humans and mice. The receptors can be classified broadly into four groups, based on the homology of their Ig-like domains and gene architecture. Receptors in the first group are characterized by the presence of the Ig constant type 2-1 (IgC2 1) and variant Ig (vlg) domains, and include the leukocyte Ig-like receptors (LILRs) and murine paired Ig-activating receptors (PIRs). The second group of receptors possess an IgC2-2 domain and comprise the killer-cell Ig-like receptors (KIRs) and platelet collagen receptor glycoprotein VI (GPVI). The third group consists of receptors with IgC2-1, and IgC2-3 or IgC2-4 domains, and includes the receptor for IgA Fc (FCAR), NKp46 and murine Ly94. The fourth group, with a single extracellular IgC2-1 domain, consists of the leukocyte-associated Ig-like receptors (LAIRs). The genomic organization of and evolutionary associations between these receptors and their domains are examined. PMID- 11929133 TI - Stat1-dependent and -independent pathways in IFN-gamma-dependent signaling. AB - The paradigm that emerged from studies during the past decade established a central role for Jak-Stat (signal transducer and activator of transcription) signaling pathways in promoting the diverse cellular responses induced by interferon gamma (IFN-gamma). However, recent studies have shown that the IFN gamma receptor activates additional signaling pathways and can regulate gene expression by Stat1-independent pathways. The diversity of gene-expression patterns mediated by Stat1-dependent and -independent mechanisms and the balance between these two pathways play an important role in the biological response to IFN-gamma. PMID- 11929132 TI - The interleukin-10 family of cytokines. AB - A family of interleukin-10 (IL-10)-related cytokines has emerged, comprising a series of herpesviral and poxviral members and several cellular sequence paralogs, including IL-19, IL-20, IL-22 [IL-10-related T-cell-derived inducible factor (IL-TIF)], IL-24 [melanoma differentiation-associated antigen 7 (MDA-7)] and IL-26 (AK155). Although the predicted helical structure of these homodimeric molecules is conserved, certain receptor-binding residues are variable and define the interaction with specific heterodimers of different type-2 cytokine receptors. This leads, through the activation of signal transducer and activator of transcription (STAT) factors, to diverse biological effects. For example, whereas IL-10 is a well-studied pleiotropic immunosuppressive and immunostimulatory cytokine, IL-22/IL-TIF mediates acute-phase response signals in hepatocytes and IL-20 induces the hyperproliferation of keratinocytes, which has been proposed as a pathogenic mechanism of psoriasis. PMID- 11929134 TI - Automating the segmentation of medical images for the production of voxel tomographic computational models. AB - Radiation dosimetry for the diagnostic medical imaging procedures performed on humans requires anatomically accurate, computational models. These may be constructed from medical images as voxel-based tomographic models. However, they are time consuming to produce and as a consequence, there are few available. This paper discusses the emergence of semi-automatic segmentation techniques and describes an application (iRAD) written in Microsoft Visual Basic that allows the bitmap of a medical image to be segmented interactively and semi-automatically while displayed in Microsoft Excel. iRAD will decrease the time required to construct voxel models. PMID- 11929135 TI - WAP-based telemedicine applications. PMID- 11929136 TI - Medical e-commerce for regional Australia. AB - The residents of rural and regional Australia have less access to health care services than in capital cities. There is a reluctance of General Practitioners to practice in the country. New information technology and government initiatives are now addressing this problem. High bandwidth videoconferencing is now being routinely used to provide psychiatric consultations to areas without this service. But this (like many other implementations of telecommunication technologies to health) has resulted in loss of revenue to regional Australia while benefiting capital cities. Thus, the current implementation of telecommunication technology to health has resulted in loss of revenue of the regions while increasing the bias towards the cities. Further, the system is not economically viable and requires the Government to inject funds for the smooth operation of the system. This paper proposes the use of telecommunication technology for enabling the communities of regional Australia to access health facilities via physical and virtual clinics. The proposed technique is self supporting and is based in the country with the intent to prevent the drain of resources from regional Australia. The technique attempts to eradicate the problem at the root level by providing a business opportunity that is based in and to cater for the needs of the remote communities. The proposed system would provide health services by physical and virtual clinics and while serving the communities would be profit centres- and thus attracting doctors and other resources to the remote communities. PMID- 11929137 TI - Wavelets for QRS detection. AB - This paper examines the use of different wavelet functions for QRS complex detection in ECG. Wavelets provide time and frequency analysis simultaneously and offer flexibility with a number of wavelet functions with different properties available. This research has examined wavelet functions with different properties to determine the effects of orthogonality and time/frequency compactness of the wavelet on the ability to correctly detect the QRS. The error in detection (false negatives and positives) is the criterion for determining the efficacy of the wavelet function. The paper reports a significant reduction in error in detection of QRS complexes with mean error reduced to 0.75%. It also reports that wavelet functions that support symmetry and compactness provide better results. PMID- 11929138 TI - Glucose dialysate induces mitochondrial DNA damage in peritoneal mesothelial cells. AB - BACKGROUND: It is known that peritoneal mesothelial cells (PMCs) are denuded in patients undergoing long-term continuous ambulatory peritoneal dialysis (CAPD); the mechanism of damage is not well known. A high quantity of glucose loaded onto PMCs in these patients may generate toxic radicals during the mitochondrial metabolism, leading to mitochondrial DNA damage that accumulates due to the incomplete repair system of this DNA. OBJECTIVE: To study damage to the PMCs of long-term CAPD patients, and to examine whether glucose overload accelerates this damage in vitro. DESIGN: Descriptive clinical and in vitro study. PARTICIPANTS: Stable CAPD patients and nonuremic patients undergoing elective abdominal surgery. METHODS: (1) Clinical Samples: 13 peritoneal tissue samples from CAPD patients and 5 omental tissue samples from patients with normal renal function were investigated. PMCs in dialysate effluent were collected from another 13 stable CAPD patients. (2) In Vitro Samples: Primary PMCs were incubated for up to 144 hours in medium containing one of the following: 5.6 mmol/L glucose (control), 56 mmol/L glucose (G), 222 mmol/L glucose (high G), or 222 mmol/L mannitol (high M; osmolar control for high G). The tissues and cells of clinical and in vitro samples were stained for light and immunoelectron microscopy with anti-8-hydroxy-2'-deoxyguanosine (anti-8-OH-dG) antibody, a marker of oxidative DNA damage. In vitro cells were also studied using transmission electron microscopy. Cellular ATP content, mitochondrial membrane potential, and intracellular generation of reactive oxygen species (ROS) were analyzed by luciferase-luciferin system, or by flow cytometry using rhodamine 123 and 2',7' dichlorodihydrofluorescein diacetate (DCFH-DA). RESULTS: Biopsy specimens showed strong cytoplasmic staining with 8-OH-dG in patients on long-term CAPD, but only faint staining in patients with end-stage renal disease before the initiation of CAPD, and no staining in patients with normal renal function. Dialysate effluent showed strong granular staining with 8-OH-dG in most PMCs in all long-term CAPD patients, but only faint and focal staining in patients at the start and after 3 5 months of CAPD. In vitro experiments also showed strong granular staining by 8 OH-dG in most PMCs cultured in high G, weak staining in G and high M, and no staining in the control. Immunoelectron microscopy revealed the localization of 8 OH-dG to mitochondria. Transmission electron microscopy showed swelling of mitochondria, with decreased cristae, in PMCs cultured in high G. However, only partial expansion of mitochondria was seen in G and high M, and no changes were seen in the control. Cellular ATP content and mitochondrial membrane potential were reduced early, followed by an increase when cultured in high G. Intracellular ROS production was also increased in PMCs cultured in high G and high M. CONCLUSIONS: These data suggest that high-glucose peritoneal dialysate may promote oxidative mitochondrial DNA damage in PMCs in CAPD patients. PMID- 11929139 TI - Laparoscopic Tenckhoff catheter implantation in children. AB - OBJECTIVE: To assess the suitability of a laparoscopic Tenckhoff catheter implantation (TCI) technique in children. DESIGN: Prospective nonrandomized controlled study. SETTING: Laparoscopic and conventional TCIs in children were performed in a tertiary-care hospital. PATIENTS: Between 1998 and 2001, 25 laparoscopic and 23 conventional TCIs were performed in 42 children. Patients in unstable clinical conditions were excluded. The laparoscope was inserted via transumbilical incision, and a forceps by percutaneous puncture. After catheter insertion, the tip was positioned in the Douglas space, and the inner cuff placed adjacent to the peritoneum, without sutures. Peritoneal dialysis was initiated immediately after surgery. MAIN OUTCOME MEASURES: Catheter-related complications during the first 4 weeks after TCI. RESULTS: After laparoscopic TCI, dialysate leakage occurred in 2 of 25 cases, one of which could be managed conservatively. In 1 patient in whom dwell volume was increased immediately after laparoscopic TCI, subcutaneous leakage occurred at the site of forceps insertion. In 2 patients with severe pre-existing intra-abdominal adhesions, outflow obstruction persisted after laparoscopic TCI. Simultaneous herniotomy was performed in 2 male infants. After conventional TCI, dialysate leakage occurred in 5 of 23 cases, 1 of which could be managed conservatively. Outflow obstruction occurred in 2 cases. CONCLUSION: Laparoscopic TCI is feasible in children of all age groups, with at least equivalent functional results compared to conventional TCI. An additional advantage is the option to identify and eliminate anatomical risk factors, such as intra-abdominal adhesions or preformed inguinal hernias in male infants. PMID- 11929140 TI - Colonization-resistant antimicrobial-coated peritoneal dialysis catheters: evaluation in a newly developed rat model of persistent Pseudomonas aeruginosa peritonitis. AB - OBJECTIVE: Development of a rat model of persistent peritonitis and evaluation of the ability of liposomal ciprofloxacin hydrogel-coated silicone to resist colonization. DESIGN: A newly developed model of persistent Pseudomonas aeruginosa peritonitis to compare the ability of liposomal ciprofloxacin hydrogel (LCH)-coated silicone versus plain silicone for resistance to bacterial colonization. ANIMALS: Male Sprague-Dawley rats. RESULTS: Inoculating the peritoneum of rats with 1 mL 0.5% agar containing 10(6) colony-forming units (cfu)/mL P. aeruginosa in the presence of a plain silicone coupon resulted in persistent peritonitis for at least 7 days. Plain silicone coupons in all 40 rats were colonized (median 2.54 x 10(3) cfu/cm2; range 5.0 x 10(1)-1.0 x 10(6) cfu/cm2) and peritoneal washings were consistently culture-positive. In contrast, the LCH coupons removed after 7 days from the 40 test rats were sterile, as were the peritoneal washings, and there was no evidence of peritonitis. Blood cultures were negative in both groups. CONCLUSIONS: Liposomal ciprofloxacin hydrogel coated silicone resists colonization in this rat model of persistent P. aeruginosa peritonitis. PMID- 11929141 TI - Influence of dialysate on gastric emptying time in peritoneal dialysis patients. AB - OBJECTIVE: Peritoneal dialysis (PD) patients frequently suffer from dyspeptic complaints such as nausea, vomiting, abdominal distension, early satiety, and anorexia. Gastroparesis might be, at least partially, a source of dyspeptic complaints in PD patients. The aim of the present study was to determine the influence of the presence and composition of dialysate on gastric emptying in PD patients. DESIGN: Prospective study. SETTING: Renal Division, Department of Internal Medicine, Ghent University Hospital, Belgium. PATIENTS: Sixty-one PD patients using different dialysate solutions, and 27 healthy volunteers. MAIN OUTCOME MEASURE: Gastric emptying of solids was assessed by the 13C-octanoic acid breath test. RESULTS: Gastric emptying was impaired in PD patients, regardless of the composition of dialysate and even if tested with an empty peritoneal cavity. Gastric emptying was significantly slower when glucose-containing dialysate was compared to an empty peritoneal cavity, or when glucose-containing dialysate was compared to icodextrin dialysate. No difference in gastric emptying could be demonstrated between glucose-containing dialysate and dialysate containing a mixture of glycerol and amino acids as osmotic agent. CONCLUSIONS: These findings suggest that the delay in gastric emptying demonstrated in the presence of peritoneal dialysate is not the consequence of a mere volume or pressure effect, but of the absorption of substrate substances with caloric and/or metabolic activity, such as glucose or glycerol and amino acids. PMID- 11929142 TI - Comparison of hemodialysis and peritoneal dialysis--a cost-utility analysis. AB - OBJECTIVE: Our aim was to compare both health-related quality of life and costs for hemodialysis (HD) and peritoneal dialysis (PD) in a defined population. DESIGN: Decision-tree modeling to estimate total costs and effects for two treatment strategies, HD and PD, among patients with chronic kidney failure, for 5 years following the start of treatment. Courses of events and health-care consumption were mapped in a retrospective matched-record study. Data on health status were obtained from a matched population by a quality-of-life questionnaire (EuroQol). The study has a societal perspective. SETTING: All dialysis departments in the southeastern health-care region of Sweden. PATIENTS: 136 patients with kidney failure, comprising 68 matched pairs, were included in a retrospective record study; 81 patients with kidney failure, comprising 27 matched triplets, were included in a prospective questionnaire study. MAIN OUTCOME MEASURES: Cost per life year and cost per quality-adjusted life year. RESULTS: The cost per quality-adjusted life year for PD was lower in all analyzed age groups. There was a 12% difference in the age group 21-40 years, a 31% difference in the age group 41-60 years, and an 11% difference in the age group 61+ years. Peritoneal dialysis and HD resulted in similar frequencies of transplantation (50% and 41%, respectively) and expected survival (3.58 years and 3.56 years, respectively) during the first 5 years after the initiation of treatment. CONCLUSION: The cost-utility ratio is most favorable for PD as the primary method of treatment for patients eligible for both PD and HD. PMID- 11929143 TI - Lack of isoprene overproduction during peritoneal dialysis. AB - OBJECTIVE: Isoprene is the constitutive unit of isoprenoid lipids and sterols. However, it is also a potential toxic and carcinogenic agent. Recent findings of a marked and prolonged isoprene overproduction induced by hemodialysis sessions raises the question of isoprene behavior in patients on peritoneal dialysis. DESIGN: A study with repeated measures per patient and healthy control. SETTING: Nephrology and Dialysis Unit and Perugia University Medical School. PATIENTS: Sixteen consecutive patients on regular continuous ambulatory peritoneal dialysis (CAPD) were evaluated. Endogenous isoprene was analyzed using gas chromatographic assay of breath isoprene, collected at set times before and after dialysis fluid exchange. RESULTS: No significant variations were found in breath isoprene concentrations in the different samples from each patient, and levels were almost stable within the normal range of healthy controls. CONCLUSION: These results show that CAPD, unlike hemodialysis, has little or no effect on isoprene and isoprenoid-related lipid turnover. This lack of increased endogenous isoprene synthesis, in addition to being a distinctive metabolic feature of CAPD, could have important pathophysiological and clinical implications. PMID- 11929144 TI - Reformulating the integrated care concept for the new millennium. AB - The concept of integrated care advocates a planned approach to predialysis care. Its components include timely referral to a renal care team, timely preparation for dialysis, and timely initiation of dialysis, with peritoneal dialysis (PD) as the initial therapy for suitable patients. This initial therapy may then be followed by timely transfer to hemodialysis (HD) when residual renal function declines or when other PD-related problems occur. We argue that at least three problems exist with the integrated care approach as it is currently being articulated. They are (1) an under emphasis on slowing the rate of progression of chronic kidney disease, aggressive management of cardiovascular risk factors and other comorbidities, and pre-emptive live donor transplantation; (2) a lack of consideration of home HD as an alternate initial therapy; and (3) a lack of discussion about patient choice. We propose a modified version of integrated care that we believe will optimize modality distribution and patient outcomes, while reducing the relative costs per patient treated, compared to the current situation in most countries. PMID- 11929145 TI - The effect of technique failure on outcome in black patients on continuous ambulatory peritoneal dialysis. AB - BACKGROUND: We previously reported that, while black patients have a better patient survival than white patients on peritoneal dialysis (PD), they also have a significantly higher technique failure rate (39% vs 8%, p < 0.0001). The purpose of this study was to determine the effect of technique failure/transfer to hemodialysis (HD) on patient survival in black PD patients. METHODS: We retrospectively evaluated 137 incident black patients entering our PD program from January 1987 to December 1997. During the course of follow-up, 82 (60%) patients remained on PD (PD group) while 55 (40%) patients were permanently transferred to HD (PD-HD group). The primary outcome measured was patient survival. RESULTS: Average age was 49 +/- 15 years, 42% were male, and 40% had diabetes mellitus. At baseline, serum creatinine was 10.8 +/- 5.4 mg/dL, serum albumin 3.4 +/- 0.7 g/dL, body mass index 27.3 +/- 6.5 kg/m2, peritoneal transport status was high in 18% and high-average in 61%, and residual glomerular filtration rate was 3.4 +/- 3.5 mL/minute. There were no significant differences in clinical features, nutritional status, peritoneal transport, residual renal function, or dialysis adequacy at baseline between the PD group and PD-HD group. While a greater proportion of patients transferring to HD had cardiac disease (53% vs 32%, p < 0.05), there were no other significant differences in 15 comorbid conditions assessed at baseline. The primary reason for transfer was peritonitis (64%) and the overall peritonitis rate in the PD-HD group was significantly higher than in the PD group (2.21 vs 1.17 episodes/patient-year, p < 0.0001). Overall follow-up was 34 +/- 25 months for PD group and 44 +/- 26 months for PD-HD group (p < 0.01), with a mean time on PD prior to transfer to HD of 22 +/- 18 months. During the course of follow-up, there were no significant differences between the two groups in the number of patients transplanted or deaths. Patient survival at 1, 2, and 5 years was 91%, 80%, and 57% for PD group and 96%, 92%, and 55% for PD-HD group [p = not significant (NS)]. A risk-adjusted time-dependent Cox regression analysis resulted in an adjusted relative risk of death that was not significantly different for those who transferred from PD to HD versus those who remained on PD (relative risk 1.49; 95% confidence interval 0.77-2.89; p = NS). CONCLUSIONS: In black patients on PD, transfer to HD is not associated with any significant difference in patient survival compared to patients remaining on PD. While a high rate of peritonitis predisposes to technique failure, we found no features at baseline predictive of patients at greatest risk to fail PD. Since technique failure does not portend a poorer prognosis, PD remains a viable option for black patients entering an end-stage renal disease program. PMID- 11929146 TI - Efficacy of a low-dose intravenous iron sucrose regimen in peritoneal dialysis patients. AB - OBJECTIVE: Sufficient iron substitution leads to a decrease in the required recombinant human erythropoietin (rHuEPO) dose and/or an increased hematocrit in dialysis patients. Intravenous (i.v.) application of larger doses of iron sucrose may be associated with hyperferritinemia, appearance of catalytically free iron, and impaired phagocyte function. Therefore, we investigated the effectiveness of a low-dose i.v. iron regimen in peritoneal dialysis (PD) patients. PATIENTS AND INTERVENTIONS: Forty-five PD patients were followed over a period of 1 year. Serum ferritin, serum transferrin saturation, and hemoglobin were measured monthly. In cases of absolute iron deficiency (serum ferritin < 100 microg/L), 50 mg iron sucrose was given i.v. every second week. In cases of functional iron deficiency (ferritin > or = 100 microg/L and transferrin saturation < 20%) and in iron repleted patients (ferritin > or = 100 microg/L and transferrin saturation > or = 20%), 50 mg i.v. iron sucrose was applied monthly. Iron therapy was stopped in cases of acute infection (until complete recovery) and when serum ferritin level was > or = 600 microg/L. RESULTS: To analyze the influence of iron substitution on erythropoiesis and rHuEPO requirements, the EPO resistance index (ERI; quotient of rHuEPO dose in units/kilogram/week and hemoglobin in grams per deciliter) was calculated every 3 months. The ERI decreased significantly during the course of the study in the whole patient group (p = 0.009) as well as in the subgroup of 21 patients with absolute iron deficiency (p = 0.01). A nonsignificant decrease in the ERI was observed within the group of 14 iron repleted patients (p = 0.5). There was no significant change in the ERI in 10 patients with functional iron deficiency (p = 0.6). CONCLUSION: The low-dose i.v. iron regimen used in this study substantially decreased rHuEPO requirements in patients with absolute iron deficiency and was effective in maintaining iron stores in iron repleted patients. However, in the absence of significant hyperparathyroidism, aluminum toxicity, or inadequate dialysis, it did not improve the ERI in patients with functional iron deficiency. PMID- 11929147 TI - A population-specific formula predicting creatinine excretion in continuous peritoneal dialysis. AB - OBJECTIVE: The Cockroft-Gault formula was shown to systematically overestimate the decline in creatinine excretion with age in continuous peritoneal dialysis (CPD) patients and is, therefore, not suitable for studying creatinine excretion. The purpose of the present study was to develop and test a population-specific formula predicting average creatinine excretion in CPD. METHODS: Creatinine excretion in urine plus dialysate was measured in 925 CPD patients. Forty patients were excluded because of evidence of noncompliance. The remaining 885 subjects were randomly grouped into a derivation group (n = 432) and a validation group (n = 453). Stepwise multiple linear regression models were used to predict creatinine excretion in the derivation group. The candidate variables, chosen because they were previously shown to be predictors of creatinine excretion in CPD, included weight (W), age (A), gender (G), diabetes (D), and interaction terms between these four variables. Estimates of creatinine excretion from the best-fit regression formula (CrExcr1) and from the Cockroft-Gault formula (CrExcr2) were compared to creatinine excretion (CrExcr) in the validation group. RESULTS: The best-fit regression model in the derivation group included all four candidate variables (W, A, G, D), but no interaction terms. This model was as follows: CrExcr1 = 302.150 - 4.380A + 171.234G - 39.041D + 11.730W (r2 = 0.477, p < 0.001). In the validation set, CrExcr = -15.795 + 0.988CrExcr1 (r2 = 0.447, p < 0.001), and CrExcr = -303.823 + 0.732CrExcr2 (r2 = 0.340, p < 0.001). When the differences between measured and predicted creatinine excretion did not take into account the sign of each individual difference, CrExcr - CrExcr1 = 201 +/- 156 mg/24 hours, and CrExcr - CrExcr2 = 235 +/- 174 mg/24 hr (p < 0.001) in the validation group. When the sign of the difference was taken into account, CrExcr CrExcr1 = -28 +/- 149 mg/24 hr, and CrExcr - CrExcr2 = 63 +/- 295 mg/24 hr (p < 0.001). CONCLUSIONS: A population-specific formula predicting creatinine excretion in CPD was derived. This formula has greater accuracy than the Cockroft Gault formula and can be used in studies of creatinine excretion in CPD. PMID- 11929148 TI - Survival of elderly patients on peritoneal dialysis: retrospective study of 292 patients, from 1982 to 1999. AB - BACKGROUND: Dialysis is becoming increasingly frequent in patients over 75 years of age. Age is a superimposed comorbid factor commonly associated with poor prognosis in these patients. OBJECTIVE: To analyze the survival of 292 patients aged over 75 years on initiation of peritoneal dialysis (PD) from September 1982 to September 1999. DESIGN: Retrospective study. SETTING: Nephrology department in a University Hospital. RESULTS: Mean age was 81.5 years (range 75-92 years); 178 patients were over 80 years and 60 patients were over 85 years. Sex ratio was 136F/156M. Ninety-day mortality rate was 12%. Excluding the first 3 months, median patient survival was 21.6 months; 226 patients died on PD and 24 were shifted to hemodialysis. Survival was inversely correlated with the Charlson combined comorbidity index (CCI), but independent of predialysis hemoglobin and serum albumin levels. Over three selected periods, 1982-1989, 1989-1995, and 1995 1999, an increase was found in mean age (79.7 +/- 3.3, 82.6 +/- 3.9, and 81.8 +/- 4.4 years; p < 0.001), CCI (7.6 +/- 1.59, 8.0 +/- 1.52, and 8.5 +/- 1.63; p = 0.01), and predialysis creatinine clearance (6.2 +/- 2.3, 6.4 +/- 2.4, and 9.8 +/ 3.8 mL/minute; p < 0.001). Median survival was similar in the various selected periods (21.0, 21.5, and 25.4 months). The incidence of peritonitis decreased from 0.63 to 0.21 episodes per patient year. CONCLUSION: From 1982 to 1999, mean age and comorbidity increased on initiation of dialysis in elderly patients, with no increase in mortality. Survival in elderly patients on PD was related to the age-comorbidity index. PMID- 11929149 TI - Polycystic kidney disease and late peritoneal leakage in CAPD: are they related? PMID- 11929150 TI - Sterile/allergic peritonitis with icodextrin in CAPD patients. PMID- 11929151 TI - Transient sterile chemical peritonitis in a CAPD patient using icodextrin. PMID- 11929152 TI - Low peritonitis rate leads to high patient survival and technique success: the first five years of a peritoneal dialysis program. PMID- 11929153 TI - Incompatibility of vancomycin and ceftazidime for intraperitoneal use. PMID- 11929154 TI - Hypotension on CAPD: can use of vasoconstrictive drugs be harmful? PMID- 11929155 TI - Uterine prolapse--a rare or rarely reported complication of CAPD? PMID- 11929156 TI - Nature's impartiality, man's inhumanity: reflections on terrorism and world crisis in a context of historical disaster. AB - This paper compares the terrorist outrages of 11 September 2001 in New York City and Washington to the Lisbon earthquake of 1 November 1755. Both events occurred, literally out of the blue, at critical junctures in history and both struck at the heart of large trading networks. Both affected public attitudes towards disaster as, not only did they cause unparalleled destruction, but they also represented symbolic victories of chaos over order, and of moral catastrophism over a benign view of human endeavour. The Lisbon earthquake led to a protracted debate on teleology, which has some parallels in the debate on technological values in modern society. It remains to be seen whether there will be parallels in the reconstruction and the ways in which major disasters are rationalised in the long term. But despite the differences between these two events--which are obviously very large as nearly 250 years of history separate them and they were the work of different sorts of forces--there are lessons to be learned from the comparison. One of these is that disaster can contribute to a perilous form of self absorption and cultural isolation. PMID- 11929157 TI - The livelihoods gap: responding to the economic dynamics of vulnerability in Somalia. AB - A 'livelihoods gap' has become evident in international aid delivery to Somalia. Existing aid interventions do not address the economic dynamics of vulnerability resulting from Somalia's long history of predatory development and asset stripping. To prevent poor households' regular return to sub-subsistence income levels after a brief period of plenty, this paper argues that aid agencies should reorient and expand existing interventions to assist poor households to capitalise on temporary improvements in environmental and security conditions. As a corollary to emergency relief and efforts to construct state institutions, it is necessary to devise country-wide interventions that will rebuild household asset bases by protecting savings during times of stress and ensuring that markets benefit poor producers. PMID- 11929159 TI - The economics of landmine clearance in Afghanistan. AB - This paper presents an economic evaluation of landmine clearance in Afghanistan. The main benefits comprise increased agricultural output, saved transport time and running costs, saved human casualties and the saved costs of supporting refugees and displaced persons. An investment of US$100 million between 1988 and 1998 is estimated to provide annual benefits of $50.3 million per annum between 1999 and 2008. This translates into net present values of between $935 and $1,744 million, depending on the rate of discount used. This contrasts with the negative NPVs estimated for several other countries. PMID- 11929158 TI - Community resilience and volcano hazard: the eruption of Tungurahua and evacuation of the faldas in Ecuador. AB - Official response to explosive volcano hazards usually involves evacuation of local inhabitants to safe shelters. Enforcement is often difficult and problems can be exacerbated when major eruptions do not ensue. Families are deprived of livelihoods and pressure to return to hazardous areas builds. Concomitantly, prevailing socio-economic and political conditions limit activities and can influence vulnerability. This paper addresses these issues, examining an ongoing volcano hazard (Tungurahua) in Ecuador where contextual realities significantly constrain responses. Fieldwork involved interviewing government officials, selecting focus groups and conducting surveys of evacuees in four locations: a temporary shelter, a permanent resettlement, with returnees and with a control group. Differences in perceptions of risk and health conditions, and in the potential for economic recovery were found among groups with different evacuation experiences. The long-term goal is to develop a model of community resilience in long-term stress environments. PMID- 11929160 TI - Earthquake drills and simulations in community-based training and preparedness programmes. AB - The San Francisco, California, bay area is subject to continuous seismic risk. One particular response has been the development of community-based training programmes designed to teach residents basic emergency response skills. Citizens are taught emergency medical techniques, search and rescue, fire suppression and other fundamental response skills. Current estimates in the Bay Area place the number of programmes at more than 100. Many programmes now include an annual community drill to reinforce the training and to evaluate the programme. The study described here is based on an evaluation of an effort initiated by BayNET (Bay Area Neighborhood Emergency Training), a voluntary association of communities with community-based disaster preparedness programmes. In April 1996, BayNET asked all of its members to hold a community earthquake drill. After the drill, a mail survey was conducted of all programme managers. The survey examined the structure and administration of the programmes, training efforts and other related components. This paper describes the typology of drill formats that communities used, the role of the simulation in the city's preparedness efforts, the qualitative costs and benefits, as well as an assessment of the drill based on survey respondents. PMID- 11929161 TI - Forced migration, processes of return and livelihood construction among pastoralists in northern Sudan. AB - The Hawaweer, a nomadic, pastoralist group in northern Sudan, were seriously affected by the drought in the Sahel during the mid-1980s. Their experience illustrates the connection between internally displaced people, normal mobility, forced migration, dilemmas and opportunities of return and how new livelihoods can be successfully constructed based on traditional rights, strong local institutions and external resources. Some displaced Hawaweer got the chance to return to their homeland as new livelihood opportunities were established; others did not get this opportunity nor would they have returned if they had been given the chance. In both situations, the processes of displacement and return had an impact on the sense of belonging and identity. PMID- 11929162 TI - A survey of international urban search-and-rescue teams following the Ji Ji earthquake. AB - On 21 September 1999, the Ji Ji earthquake killed 2,347 people. In the immediate aftermath the international community, mobilised rapidly and sent urban search and-rescue (US&R) teams to the scene. This paper will present an annotated survey of the expertise and standard of equipment of international US&R teams following that earthquake which could serve as a blueprint for the establishment of US&R teams elsewhere at risk from earthquakes. PMID- 11929163 TI - Sensory, chemical and bacteriological changes in vacuum-packed pressurised squid mantle (Todaropsis eblanae) stored at 4 degrees C. AB - Sensory, chemical and bacteriological changes were studied in vacuum-packed squid mantles (Todaropsis eblanae) that were pressurised at 150, 200, 300 and 400 megapascal (MPa) for 15 min at ambient temperature and stored at 4 degrees C. Sensory analysis showed that the higher the pressurisation the longer the shelf life. Thus, the lot pressurised at 400 MPa was rejected after 28 days' storage compared with 7 days for the untreated lot. The chemical results generally corresponded with the sensory ones. Furthermore, ammonia (NH3) and trimethylamine (TMA) were produced in the pressurised lots after a pressure dependent delay. Urea decreased to low levels in all lots with the exception of the 400 MPa lot. Onset of production of agmatine, the dominant amine in this species, and other biogenic amines was delayed by increasing pressure, but still, high concentrations of these amines were detected in pressurised lots of acceptable sensory quality. Microbial counts conducted after 1 day of storage showed that the bacterial load was reduced by all pressures, reaching levels below the detection limit in the lots treated with 200-400 MPa. However, growth was resumed in all lots after a pressure dependent delay. Luminous bacteria predominated initially in the lots pressurised at 300 and 400 MPa, but were outnumbered by Enterobacteriaceae and lactic acid bacteria at the time of sensory rejection of these lots. All colonies isolated prior to pressurisation were identified as Photobacterium phosphoreum. This bacterium also resumed growth faster than other members of the endogenous microflora after pressurisation. All luminous colonies were identified as P. phosphoreum. Lactic acid bacteria isolated at the final sampling point of the lot pressurised at 400 MPa were identified as Carnobacterium piscicola and Carnobacterium divergens, while Serratia liquefaciens and Proteus vulgaris make up Enterobacteriaceae. PMID- 11929164 TI - Antimicrobial activity of individual and mixed fractions of dill, cilantro, coriander and eucalyptus essential oils. AB - Essential oils from dill (Anethum graveolens L.), coriander (seeds of Coriandrum sativum L.), cilantro (leaves of immature C. sativum L.) and eucalyptus (Eucalyptus dives) were separated into heterogeneous mixtures of components by fractional distillation and were analyzed by gas chromatography-mass spectroscopy. Minimum inhibitory concentrations against gram-positive bacteria, gram-negative bacteria and Saccharomyces cerevisiae were determined for the crude oils and their fractions. Essential oil of cilantro was particularly effective against Listeria monocytogenes, likely due to the presence of long chain (C6-C10) alcohols and aldehydes. The strength and spectrum of inhibition for the fractions often exceeded those determined in the crude oils. Mixing of fractions resulted in additive, synergistic or antagonistic effects against individual test microorganisms. PMID- 11929165 TI - Microbiological and organoleptic qualities of vacuum-packaged ground beef prepared from pasteurized manufacturing beef. AB - Manufacturing beef was pasteurized by immersion in water of 85 degrees C for 60 s, or was not pasteurized before being coarsely ground. The coarsely ground beef was vacuum-packaged and stored at 2 degrees C for up to 6 weeks. Before storage and at weekly intervals, three packs of pasteurized and three of unpasteurized beef were opened. The meat from each pack was finely ground and displayed in two overwrapped packs at 4 degrees C for 3 days after storage for up to 3 weeks, or for 2 days after longer storage times. Samples for microbiological analysis were obtained at the times of preparation of display packs and at the end of display. Displayed meat was assessed daily for colour, discolouration and retail acceptability, and for odour intensity and acceptability at the end of display. Before storage, the numbers of total aerobic bacteria, presumptive pseudomonads and presumptive Brochothrix thermosphacta recovered from pasteurized meat were >1 log unit less than the corresponding numbers recovered from unpasteurized meat, but the numbers of presumptive lactic acid bacteria and presumptive enterobacteria were <1 log unit less from pasteurized than from unpasteurized meat. After all periods of storage and display, the numbers of bacteria recovered from pasteurized and unpasteurized meat at each time were mostly similar. The colour of pasteurized meat was perceived as being paler than that of unpasteurized meat, but discolouration and retail acceptability of pasteurized and unpasteurized meat were perceived as similar at most times. The odours of displayed, pasteurized and unpasteurized meat were perceived to be similar at all times that odours were assessed. The findings indicate that pasteurizing of manufacturing beef to improve the microbiological safety of ground beef would not unacceptability degrade the appearance of the ground product, but that the storage life of ground beef would not be greatly extended by the treatment. PMID- 11929167 TI - Detection of Bacillus cereus in foods by colony hybridization using PCR-generated probe and characterization of isolates for toxins by PCR. AB - Isolates of Bacillus cereus from traditional Indian foods were detected by colony hybridization using the PCR-generated phospholipase (PL-1) probe. In all, 29 isolates picked up by the probe were confirmed as B. cereus by conventional cultural and biochemical characteristics. All the isolates reacted positively in PCR with phospholipase (PL-1) primers. Among the native isolates, 11 of them showed the discontinuous pattern of haemolysin BL activity in gel diffusion assay. Though 14 isolates reacted positively in PCR with primers (Ha-1) specific to the B gene of haemolysin BL, only four of them showed both the presence of gene and haemolysin BL activity. More than 50% of the isolates indicated their potential enterotoxigenicity by reacting positively with primers specific for the BceT gene encoding for diarrhoeal enterotoxin. PCR with primers for different inverse (IS) repeat elements revealed that isolates carrying transposon IS 231-P 231-1 did not carry IS 240-P 240. Some of the isolates were devoid ofthese IS elements. The study demonstrated the potential of using of a PCR-generated labelled PL-1 probe for the direct detection of B. cereus in food samples and PCR for characterizing the toxigenic isolates. PMID- 11929166 TI - Prevalence of enterovirus and hepatitis A virus in bivalve molluscs from Galicia (NW Spain): inadequacy of the EU standards of microbiological quality. AB - A study of the presence of hepatitis A virus (HAV) and enterovirus (EV) in shellfish from the northwestern coast of Spain, one of the most important mussel producers in the world, was carried out employing dot-blot hybridization and RT PCR techniques. In addition, bacterial contamination of the samples was evaluated by Escherichia coli (EC) counts, according to the European Union (EU) standards of shellfish microbiological quality. Shellfish samples included raft-cultured and wild mussels, as well as wild clams and cockles. Bacterial counts showed that the majority of samples (40.8%) could be classified as moderately polluted following the EU standards, and therefore should undergo depuration processes. However, differences in bacterial contamination were observed between cultured mussel and wild shellfish. Thus, percentage of clean samples (<230 EC/100 g shellfish) was clearly higher in cultured mussels (49.1%) than in wild mussels (22.8%) or clams and cockles (10.7%). HAV was detected in 27.4% and EV in 43.9% of the samples that were analyzed. Simultaneous detection of both viral types occurred in 14.1% of the samples. Statistical tests of dependence (chi-square test) showed no relationship either between viral and bacterial contamination, or between the presence of HAV and EV. Comparative analysis of hybridization and RT PCR for viral detection yielded different results depending on the virus type that was studied, RT-PCR being effective for HAV but not for EV detection. The obtained results reinforce once again the inadequacy of bacteriological standards to assess viral contamination and suggest that although virological analysis of shellfish is possible by molecular techniques, interlaboratory standardization and validation studies are needed before the routine use in monitoring shellfish microbiological safety. PMID- 11929168 TI - Evaluation of sorbitol-salicin MacConkey medium containing cefixime and tellurite (CT-SSMAC medium) for isolation of Escherichia coli O157:H7 from raw vegetables. AB - The utility of CT-SSMAC medium (sorbitol-salicin MacConkey medium containing cefixime and tellurite) for the isolation of Escherichia coli O157:H7 from raw vegetables was investigated. The colonies of all E. coli O157:H7 and O157:NM strains tested were colorless and beta-galactosidase-positive on CT-SSMAC medium. Furthermore, the number of colorless colonies on the CT-SSMAC medium was less than that on the sorbitol MacConkey medium containing cefixime and tellurite (CT SMAC medium) from several raw vegetable samples. All colorless colonies grown on CT-SSMAC medium from raw vegetable samples were beta-galactosidase-negative. These findings suggest that the CT-SSMAC medium is useful for the isolation of E. coli O157:H7 from raw vegetable samples. PMID- 11929169 TI - Comparison of methods for fluorescent detection of viable, dead, and total Escherichia coli O157:H7 cells in suspensions and on apples using confocal scanning laser microscopy following treatment with sanitizers. AB - The influence of treating Escherichia coli O157:H7 cells labeled with an enhanced green fluorescent protein (EGFP) plasmid with 20 microg/ml active chlorine, 100 mg/ml hydrogen peroxide, and 80 mg/ml acetic acid on fluorescence intensity was determined. In addition, fluorescent staining methods to differentiate viable and dead E. coli O157:H7 cells on the cuticle of Red Delicious cv. apples following treatment with water or 200 microg/ml active chlorine were evaluated. Suspensions of E. coli O157:H7 EGFP+ cells were exposed to chemical treatment solutions for 0, 30, 60, 120, or 300 s before populations (log10 cfu/ml) were determined by surface plating, and fluorescence intensities of suspensions and individual cells were measured using spectrofluorometry and confocal scanning laser microscopy (CSLM), respectively. The relative fluorescence intensity of suspensions and individual cells changed upon exposure to various treatments. Results indicate that the use of EGFP to tag E. coli O157:H7 may not be appropriate for investigations seeking to microscopically differentiate viable and dead cells on produce following surface treatment with sanitizers. SYTOX Orange and SYTOX Green nucleic acid stains fluorescently labeled dead E. coli O157:H7 cells attached to apple cuticles more intensely than did propidium iodide. A cross-signal occurred between CSLM photomultipliers when examining tissues treated with SYTOX Orange to detect dead cells and antibody labeled with Alexa Fluor 488 to detect total (dead and viable) cells. Because of the possibility of cross-signal resulting in an overestimation of the number of dead cells on apples and, perhaps, other produce treated with these stains, SYTOX Green is preferred to detect dead cells and antibody labeled with Alexa Fluor 594 is preferred to detect the total number of cells on apple surfaces following treatment with sanitizers. The performance of SYTOX Green in combination with Alexa Fluor 594 to detect dead and total cells of E. coli O157:H7 on other produce remains to be determined. PMID- 11929171 TI - Antibacterial activity of chitosans and chitosan oligomers with different molecular weights. AB - Antibacterial activities of six chitosans and six chitosan oligomers with different molecular weights (Mws) were examined against four gram-negative (Escherichia coli, Pseudomonas fluorescens, Salmonella typhimurium, and Vibrio parahaemolyticus) and seven gram-positive bacteria (Listeria monocytogenes, Bacillus megaterium, B. cereus, Staphylococcus aureus, Lactobacillus plantarum, L. brevis, and L. bulgaricus). Chitosans showed higher antibacterial activities than chitosan oligomers and markedly inhibited growth of most bacteria tested although inhibitory effects differed with Mws of chitosan and the particular bacterium. Chitosan generally showed stronger bactericidal effects with gram positive bacteria than gram-negative bacteria in the presence of 0.1% chitosan. The minimum inhibitory concentration (MIC) of chitosans ranged from 0.05% to >0.1% depending on the bacteria and Mws of chitosan. As a chitosan solvent, 1% acetic acid was effective in inhibiting the growth of most of the bacteria tested except for lactic acid bacteria that were more effectively suppressed with 1% lactic or formic acids. Antibacterial activity of chitosan was inversely affected by pH (pH 4.5-5.9 range tested), with higher activity at lower pH value. PMID- 11929170 TI - Comparison of aqueous commercial cleaners for effectiveness in removing Escherichia coli O157:H7 and Salmonella muenchen from the surface of apples. AB - Unpasteurized fruit juice and cider have been implicated in outbreaks of Escherichia coli O157:H7 and Salmonella infections, yet various processes used to clean and sanitize fruits before producing juice have not been thoroughly studied for their effectiveness in removing pathogens. The objective of this study was to evaluate cleaners used in the apple industry for their efficacy in removing E. coli O157:H7 and Salmonella from the surface of apples. E. coli O157:H7 was transformed with green fluorescent protein plasmid (pGFP). In addition to encoding for the production of GFP, the plasmid also encodes for ampicillin resistance. S. muenchen was adapted to grow in media containing 50 microg/ml nalidixic acid. The use of ampicillin and nalidixic acid resistant strains enabled enumeration of the pathogen without interference by microflora naturally present on apples. Unwaxed Red Delicious cv. apples were surface inoculated with 8.58 log10 cfu of E. coli O157:H7 and 8.11 log10 cfu of S. muenchen. Five commercial apple cleaners were applied at concentrations and exposure times recommended by manufacturers. Populations of E. coli O157:H7, S. muenchen, aerobic mesophiles, and yeasts and molds on apples treated with cleaners and water (control) were determined. Compared to washing with water, treatment with cleaners removed or killed up to 2.86, 3.11, 2.48, and 0.73 log10 cfu of E. coli O157:H7, S. muenchen, aerobic mesophiles, and yeasts and molds per apple, respectively. There were differences in the effectiveness of cleaners in removing pathogens, but pH (2.0 and 12.0) and concentration (1% and 5%) of cleaner, and time of exposure (0.5-2 min) were not correlated with magnitude of reduction in population. The use of some types of cleaners commercially formulated for apples may contribute significantly in attaining target 5-log10 reductions of pathogens on the fruit intended for unpasteurized juice production or the fresh produce market. PMID- 11929172 TI - Thermal inactivation of poliovirus type 1 in water, milk and yoghurt. AB - Loss of infectivity of poliovirus type 1, strain Sabin, during heating, freezing, and storage in water, milk and yoghurt was determined by plaque-titration in Vero cell cultures. The heating experiments simulated the conditions arising during the processing of milk and yoghurt, for example high-temperature heating (95 degrees C, 15 and 30 s), short-time pasteurization (72 degrees C, 15 and 30 s), long-time pasteurization (62 degrees C, 30 min), and yoghurt-fermentation (42 degrees C, 30 min and 180 min). Only high-temperature heating, long-time pasteurization and short-time pasteurization for 30 s proved to be reliable methods of inactivating polioviruses present in water, milk and yoghurt completely. Short-time pasteurization for 15 s and the conditions of yoghurt fermentation failed to cause complete inactivation of polioviruses. Additionally, polioviruses mixed in milk or yoghurt withstood these procedures with significantly lower reductions of infectivity than in water. Heating at 55 degrees C for 30 min resulted in complete inactivation of polioviruses, regardless of the suspending medium. The infectivity of polioviruses is scarcely affected by freezing (-20 degrees C, 30 min) and storage (24 days) at low temperatures (4 degrees C) and high humidity (a(w) = 0.99). PMID- 11929173 TI - Survival of bifidobacteria after spray-drying. AB - To investigate the survival of bifidobacteria after spray-drying, Bifidobacterium infantis CCRC 14633, B. infantis CCRC 14661, B. longum ATCC 15708, B. longum CCRC 14634 and B. longum B6 were first spray-dried with different carrier media including 10% (w/w) gelatin, gum arabic and soluble starch. B. infantis CCRC 14633 and B. longum were also determined in skim milk. It was found that survival of bifidobacteria after spray-drying varied with strains and is highly dependent on the carriers used. Among the test organisms, B. longum B6 exhibited the least sensitivity to spray-drying and showed the highest survival of ca. 82.6% after drying with skim milk. Comparisons of the effect of carrier concentrations revealed that spray-drying at 10% (w/w) gelatin, gum arabic or soluble starch resulted in the highest survival of bifidobacteria. In addition, among the various outlet-air temperatures tested, bifidobacteria showed the highest survival after drying at 50 degrees C. Elevation of outlet-air temperature caused increased inactivation of bifidobacteria. However, the inactivation caused by increased outlet-air temperature varied with the carrier used, with the greatest reduction observed using soluble starch and the least with skim milk. PMID- 11929174 TI - Change in the composition of the microflora on vacuum-packaged beef during chiller storage. AB - Five fresh beefcut samples were divided into 10 pieces, respectively, vacuum packaged and stored at 2 degrees C for up to 6 weeks. The average pH values of five pieces from five beef samples were 5.62 +/- 0.04 at the start of storage and 5.12 +/- 0.07 after 6 weeks of storage. The pieces were homogenized, diluted and cultivated at weekly intervals on glucose-blood-liver (BL), de Man Rogosa and Sharpe (MRS) and trypticase soya (TS) agars at 7 degrees C for 10-14 days. From plates with 30-300 colonies, or the highest number if below 30, 15 colonies were randomly picked from each of the three media used for each beef sample. Based on morphologies, SDS-PAGE whole-cell protein profiles and physio-chemical characteristics, a total of 1493 strains isolated were identified as Brochothrix thermosphacta (64), Carnobacterium divergens (79), Carnobacterium piscicola (27), Lactobacillus algidus (637), Lactobacillus sp. (4), Lactococcus piscium (270), Leuconostoc gelidum (375), Acinetobacter (3), Aeromonas (1), Bacillus (10), Corynebacterium (3), Enterobacteriaceae (1), Pseudomonas (13) or Psychrobacter (6). A wider range of organisms was isolated from TS (13 organisms) and BL (7) agars than from MRS agar (3). Leuc. gelidum, Lc. piscium and L. algidas increased in numbers during the first 3 weeks of storage from about 5 x 10(3) cfu/g to the level of about 10(8) cfu/g and persisted at this level thereafter. C. divergens and C. piscicola were inconsistently detected, but seemed to persist at the relatively low level of about 5 x 10(7) cfu/g during the last 3 weeks of storage. B. thermosphacta increased to the level of about 7 x 10(5) cfu/g during the first 2 weeks of storage, but was not detected thereafter. Remaining organisms were detected sporadically at levels of <3.5 x 10(2) cfu/g during the first 2 weeks of storage. PMID- 11929175 TI - Atopy and neural damage. AB - The occurrence of myelitis with atopic diathesis (atopic myelitis) affecting young adults has recently been noted in Japan. The disease preferentially affects the posterior column of the cervical spinal cord, as shown clinically and by MRI. It is characterized by hyperIgEaemia and the presence of mite antigen-specific IgE. The spinal cord lesions have been shown to be eosinophilic inflammation on biopsy and thus an allergic mechanism is thought to be operative in this condition. In addition, we also found that Hirayama disease, juvenile muscular atrophy of the distal upper extremity, is also associated with airway allergy such as allergic rhinitis and atopic asthma. In children, poliomyelitislike illness after acute asthma attacks is well known as Hopkins syndrome. Moreover, by the prospective study of the history of allergic disorders in common neurologic diseases, an association between spinal progressive muscular atrophy (SPMA) and asthma as well as between myelitis and atopic dermatitis has been demonstrated. These observations strongly suggest a link between atopic diathesis and spinal cord damage. Central nervous system damage associated with atopic diathesis may be classified into two types; eosinophilic myelitis preferentially affecting the cervical spinal cord and lower motor neuron damage, such as Hopkins syndrome, Hirayama disease and SPMA. The former is typically associated with atopic dermatitis while the latter, with airway allergy. PMID- 11929176 TI - Current therapy of multiple myeloma. AB - Not all patients who fulfill the minimal criteria for the diagnosis of multiple myeloma should be treated. If a patient is younger than 70 years, autologous peripheral blood stem cell transplantation should be seriously considered. Major challenges for stem cell transplantation are: 1) the inability to eradicate multiple myeloma from the patient, and 2) removal of myeloma cells and their precursors from the reinfused stem cells. Allogeneic transplantation cannot be recommended at present because of the excessive mortality. Nonmyeloablative approaches are promising. There is no evidence that combinations of alkylating agents are superior to melphalan and prednisone. The use of thalidomide and intermittently administered prednisone for maintenance is being explored. New agents include the immunomodulatory drugs, inhibitors of the ubiquitin proteasone pathway such as PS-341, antiangiogenesis drugs including 2-methoxy-estradiol, and farnesyl transferase inhibitors. Management of skeletal complications, hypercalcemia, anemia, infection, spinal cord compression, and renal insufficiency is discussed. PMID- 11929177 TI - Elevated plasma procoagulant and fibrinolytic markers in patients with chronic obstructive pulmonary disease. AB - OBJECTIVE: There is clinical and pathological evidence of thrombosis in pulmonary vessels of patients with chronic obstructive pulmonary disease (COPD). The purpose of this study was to investigate the presence of hypercoagulability and determine the extent of this abnormality in COPD patients. PATIENTS AND METHODS: We measured plasma levels of thrombin antithrombin III complex (TAT), fibrinopeptide A (FPA), tissue plasminogen activator-plasminogen activator inhibitor (tPA-PAI): markers of coagulation-fibrinolysis-system, and also beta thromboglobulin (beta-TG): a marker of platelet activation, in 40 COPD patients and in 20 control subjects. Measurements were also repeated 12 months after entry in all patients. RESULTS: TAT, FPA, tPA-PAI, and beta-TG concentrations were significantly higher in COPD than in control subjects. At 12 months follow-up, deltaA-aDO2 and delta%FEV1 were significantly higher in patients with high TAT or tPA-PAI levels than in patients with low levels and TAT, FPA and tPA-PAI levels remained elevated, although beta-TG levels decreased after domiciliary O2 therapy. CONCLUSION: Our results showed an enhanced prothrombotic process in COPD patients, which could potentially account for the increased thrombosis in pulmonary vessels in these patients. PMID- 11929178 TI - Glucose and oxygen hypometabolism in aceruloplasminemia brains. AB - OBJECTIVE: Aceruloplasminemia is an iron metabolic disorder caused by mutations in the ceruloplasmin gene. It is characterized by progressive neurodegeneration in association with iron accumulation. Excess iron functions as a potent catalyst of biologic oxidation. Previously we showed that an increased iron concentration is associated with the products of lipid peroxidation in the serum, cerebrospinal fluid, and brain tissues. To clarify the free radical-mediated tissue injury caused by intracellular iron accumulation through mitochondrial dysfunction. PATIENTS AND METHODS: We have measure brain oxygen and glucose metabolisms using positron emission tomography (PET) and examined brains at autopsy for iron contents and activities of the mitochondrial respiratory chain in two affected patients who had different truncation mutations of the ceruloplasmin gene. RESULTS: PET showed a marked decrease in glucose and oxygen consumption in the entire brain of aceruloplasminemia patients, with a preponderance of metabolic reduction in basal ganglia. Enzyme activities in the mitochondrial respiratory chain of the basal ganglia were reduced to approximately 45% and 42% respectively for complexes I and IV. An inverse relationship was shown between the amounts of iron accumulated and the levels of mitochondrial enzyme activities in all the brain regions examined. CONCLUSION: Iron-mediated free radicals may contribute to the impairment of mitochondrial energy metabolism in aceruloplasminemia. PMID- 11929179 TI - Hepatic angiomyolipoma associated with splenic hamartoma. AB - A 52-year-old woman was admitted to our hospital with thrombophlebitis of the internal jugular vein. Abdominal ultrasonography demonstrated a high echogenic mass measuring 4.5 cm in diameter in the liver, and abdominal CT revealed another liver tumor and an isodensity mass in the spleen. Abdominal MRI and angiography were performed and we presumed the tumors to be two hepatic angiomyolipoma and a splenic hamartoma. As an abdominal CT 21 months later revealed that all tumors were growing, these tomors were surgically resected. The histological diagnoses were hepatic angiomyolipoma and splenic hamartoma. PMID- 11929180 TI - Unusual swallow syncope caused by huge hiatal hernia. AB - Swallowing is a rare cause of syncope. A 76-year-old woman was admitted to a hospital due to postprandial loss of consciousness. Although no remarkable cardiogenic problem was proven, upper gastrointestinal barium examination revealed a huge hiatal hernia. Both echocardiography and MRI presented the collapsed left atrium due to the herniated stomach. Water pouring examination successfully represented lightheadedness, and Nissen's fundoplication was carried out. After this procedure, she never suffered from syncopal attack. PMID- 11929181 TI - Interferon therapy for hepatitis C virus (HCV)-related advanced chronic hepatitis after treatment of recurrent hepatocellular carcinoma: a successful case. AB - A complete response was obtained by interferon (IFN) therapy for hepatitis C virus (HCV)-related advanced chronic hepatitis after curative treatment of initial and recurrent hepatocellular carcinoma (HCC). The patient is alive 73 months after diagnosis of initial HCC and is in a tumor-free state 53 months after diagnosis of the last HCC. This case suggest that it is possible to suppress recurrence and to prolong the life of patients with HCV-related advanced chronic hepatitis, if a complete response is obtained by means of IFN therapy after the curative treatment of the recurrent HCC. PMID- 11929182 TI - Swallowing syncope: complex mechanisms of the reflex. AB - A 69-year-old woman was admitted to our hospital for the examination of syncope. When she ate solid food, she had dizziness or loss of consciousness. The ambulatory ECG suggested sino-atrial block during swallowing with a maximum sinus pause of 6 seconds. An electrophysiologic study revealed pre-existing sinus node dysfunction, which was exaggerated by the balloon inflation in the esophagus. Atropine counteracted the slowing of the basal sinus rate induced by esophageal pressure, but it did not block the effect on the maximum sinus node recovery time. This observation suggested that the syncope was mediated partly by a non vagal mechanism. PMID- 11929183 TI - Systemic capillary leak syndrome. AB - A 40-year-old woman was referred to our hospital with severe hypovolemic shock and anasarca. The laboratory findings showed marked hemoconcentration and a decrease in total serum protein with the presence of monoclonal IgG-lambda. She had had a similar episode of generalized edema 2 years previously. We diagnosed the patient as having typical systemic capillary leak syndrome (SCLS) and she improved gradually after infusion of albumin-containing fluid. SCLS is a very rare condition caused by unexplained episodic capillary hyperpermeability. Its treatment has remained largely supportive and the prognosis is generally poor. Awareness of SCLS is necessary for improvement of the outcome. PMID- 11929184 TI - Microvascular angina accompanied by epicardial coronary artery spasm. AB - A case of microvascular angina accompanied by coronary artery spasm is described. A 54-year-old woman had anginal pain at rest and during exercise. Both exercise testing and rapid atrial pacing caused significant ST depression in the inferior and all precordial leads. Exercise thallium myocardial scintigraphy was negative despite similar ST depressions. Coronary angiography revealed insignificant stenoses of the left anterior descending coronary artery after the injection of nitrate. Intracoronary acetylcholine provoked diffuse spasm of the artery with concurrent myocardial lactate production. Coronary flow reserve assessed with papaverine was 2.75. The combined use of amlodipine and high-dose nicorandil was effective for the treatment of angina. PMID- 11929185 TI - End-stage renal disease associated with familial Mediterranean fever. AB - A 39-year-old man had been suffering from periodic fever since childhood. He was started on hemodialysis due to secondary amyloidosis on December 2000. The patient was believed to have Familial Mediterranean fever (FMF) because of recurrent fever with peritonitis, arthritis and inflammatory changes and secondary amyloidosis in his kidneys, heart and colon. No other family member had recurrent fever. IL-6, TNF, and dopamine beta-hydroxylase were not increased in the febril phase. The patient was homozygous for the M6941 mutation. We report the first Japanese case of FMF associated with amyloidosis and confirmed by a gene mutation. PMID- 11929186 TI - Treatment of bronchorrhea by corticosteroids in a case of bronchioloalveolar carcinoma producing CA19-9. AB - A case of gastrointestinal cancer-associated antigen (CA19-9)-positive bronchioloalveolar carcinoma accompanied by bronchorrhea and respiratory failure successfully treated with corticosteroids is reported. The patient was treated with pulse methylprednisolone at a dose of 1,000 mg/day for three days, followed by oral prednisolone (60 mg/day). Within 2 days, the sputum volume decreased from >100 ml/day to 20 ml/day and it was finally controlled to 0-10 ml/day. The reduction in the sputum volume was associated with alleviation of dyspnea and hypoxemia. The levels of CA19-9 in the serum and the sputum were extremely high and an immunocytochemical study showed that the tumor cells were stained by CA19 9 antibody. This case demonstrates the therapeutic value of corticosteroids in the treatment of bronchorrhea in subjects with bronchioloalveolar carcinoma. PMID- 11929187 TI - Corticosteroid therapy for hemolytic anemia and respiratory failure due to Mycoplasma pneumoniae pneumonia. AB - This is a report of hemolytic anemia and respiratory failure due to Mycopkisma pneumoniae pneumonia. His chest CT scans showed bilateral diffuse thickened bronchovascular bundles and emphysematous changes. The pulmonary function test supported the diagnosis of chronic obstructive pulmonary disease (COPD). He was diagnosed as coldagglunitin-associated hemolytic anemia and M. pneumonzae pneumonia in inapparent COPD. Corticosteroid administration was remarkabily effective for hemolytic anemia and beneficial for acute exacerbation of COPD. PMID- 11929188 TI - An autopsy case of POEMS syndrome with a high level of IL-6 and VEGF in the serum and ascitic fluid. AB - A 45-year-old woman was hospitalized because of systemic edema and peripheral nerve impairment. The patient had complications of organomegaly, endocrinopathy, and monoclonal gammmopathy, and was diagnosed with POEMS syndrome based on these characteristic signs and symptoms. Interleukin-6 (IL-6) and vascular endothelial growth factor (VEGF) levels in the serum and ascitic fluid were high. Many of the patient's symptoms were ameliorated, and IL-6 and VEGF levels in the serum and ascitic fluid decreased slightly during chemotherapy, but she died of respiratory failure. Autopsy revealed severe systemic edema and macroscopic hemorrhage in many organs, but VEGF and IL-6 producing cells were not found. PMID- 11929189 TI - Unusual fatty infiltration of the soleus muscle in a patient with congenital nemaline myopathy: fat-suppression muscle MRI. AB - A 27-year-old man developed gait disturbance over a three-year period. Histochemical analysis suggested nemaline bodies, type I fiber atrophy and type II fiber hypertrophy. Conventional magnetic resonance imaging (MRI) showed a severe degree of homogenous hyperintensity in the soleus muscle. Fat-suppression MRI exhibited marked diffuse hypointensity in the soleus muscle. MRIs demonstrated unusual fatty proliferation of the soleus muscle. Fat-suppression MRI of skeletal muscles is beneficial in evaluating the accurate topography and degree of fatty infiltration. Radiological patterns of muscle damage are variable in patients with congenital nemaline myopathy, similar to the heterogeneous clinical aspects. PMID- 11929190 TI - An autopsy case of intravascular lymphomatosis with dermatomyositis. AB - A 62-year-old man was admitted to hospital with general malaise, arthralgia, edema, and high-grade fever. He was diagnosed as dermatomyositis because of typical skin rashes and muscle weakness. His symptoms were improved by high-doses of prednisolone and cyclosporin A, with the exception of the skin rash over the back. High-grade fever developed again when tapering prednisolone. His condition deteriorated gradually. Paresthesia, hypothyroidism, metabolic acidosis, and disseminated intravascular coagulopathy occurred, and he died eight months after the first admission. Postmortem examination revealed intravascular proliferation of atypical mononuclear cells in the lumens of small vessels in all organs. Intravascular lymphomatosis (B cell type) was diagnosed. PMID- 11929191 TI - Monckeberg's arteriosclerosis. PMID- 11929192 TI - Uncoupling proteins--a new family of proteins with unknown function. AB - Uncoupling proteins are inner mitochondrial membrane proteins, which dissipate the proton gradient, releasing the stored energy as heat. Five proteins have been cloned, named UCP1, UCP2, UCP3, UCP4 and UCP5/BMCP1. These proteins are structurally related but differ in tissue expression. UCP1 is expressed uniquely in the brown adipose tissue, while UCP2 is widely distributed, UCP3 is mainly restricted to skeletal muscle and UCP4 and UCP5/BMCP1 expressed in the brain. The properties and regulation of the uncoupling proteins and their exact function has been the focus of an intense research during recent years. This review briefly summarizes the actual knowledge of the properties and function of this new family of proteins. While UCP1 has a clear role in energy homeostasis, the newcomers UCP2-UCP5 may have more delicate physiological importance acting as free radical oxygen scavengers and in the regulation of ATP-dependent processes, such as secretion. PMID- 11929194 TI - Neuropeptide Y fails to normalize food intake in zinc-deficient rats. AB - Zinc deprivation results in decreased and cyclic food intake in rats. We determined the response of zinc-deprived rats to neuropeptide Y (NPY). In a preliminary experiment, rats were fed a low (-Zn; <1 mg/kg) or adequate zinc diet (+Zn; 100 mg/kg) for 4 days. NPY (5 or 10 microg) was then administered via an intracerebroventricular (ICV) cannula and food intake measured for 4 h. NPY stimulated food intake in all rats, but the difference in food intake due to zinc deprivation persisted. In a subsequent experiment, rats were fed the low zinc and adequate zinc diets for 4, 5 or 6 days. Food intake was suppressed in rats fed the low zinc compared to the adequate zinc diet on all of these days. When NPY (10 microg) was administered at the onset of the light cycle, the food intake was approximately 2.5-fold greater regardless of dietary zinc status, but the amount of food consumed by rats fed low zinc was approximately one-half the quantity consumed by NPY-stimulated zinc-adequate rats. NPY administered at the onset of dark failed to stimulate food intake in either dietary group although the total intake difference due to zinc status persisted. ICV administration of 5 nmol of zinc prior to NPY injection failed to correct the food intake response of the zinc-deficient rats. We conclude that the basis of the reduced food intake of zinc-deficient rats does not relate to NPY quantity or release, or to impairment of its signal transduction. There appears to be another undefined factor that limits food intake in zinc deficiency. PMID- 11929193 TI - Changes in UCP2, PPARgamma2, and c/EBPalpha gene expression induced by a neuropeptide Y (NPY) related receptor antagonist in overweight rats. AB - Neuropeptide Y (NPY), a peptide released by nervous cells, appears to contribute to adiposity regulation by increasing food intake and inhibiting lipolysis. New NPY receptor related antagonists such as S.A.0204 are being developed as potential anti-obesity drugs affecting adipocyte lipid metabolism and thermogenesis. In this sense, those animals fed on a high-energy yielding (cafeteria) diet decreased body fat weight as compared to overweight controls, when they were administered with S.A.0204, and increased body temperature, which statistically correlated with high UCP2 mRNA expression levels in white adipose tissue. In addition, the in vivo NPY-antagonist administration was able to prevent white adipose tissue growth in animals fed the cafeteria (high-fat) diet by impairing PPARy and CIEBPalpha mRNA expression in white fat cells. In summary, this novel NPY related-antagonist S.A.0204 may regulate body fat deposition by affecting both energy dissipation and white adipose tissue deposition, representing a potential new pharmacological strategy for obesity management. PMID- 11929195 TI - Differential involvement of dopamine D1 and D2 receptors and inhibition by dopamine of hypothalamic VMN neurons in early postnatally overfed juvenile rats. AB - Dopamine is among the neurotransmitters involved in central regulation of food intake, and body weight control. To study possible changes in neuronal responses to dopamine, single unit activity of the ventromedial hypothalamic nucleus (VMN) was recorded in brain slices of normal and obese rats. The latter had developed overweight throughout juvenile life (p < 0.05) by early postnatal over nourishment due to a reduction of litter size from 3rd to 21st day of life (small litters, SL). With effective concentrations of about 100-500 nM/I dopamine inhibited significantly more VMN neurons in obese than normal rats (Chi-square p < 0.05). While D2 receptors in the VMN are reported to mediate inhibition of food intake, the responses to dopamine were blocked by D2 receptor antagonists in significantly fewer neurons of SL than normal rats (p < 0.05). Furthemore, including results of action of D1 receptor agonists we found that significantly more neurons in SL than NL rats seem to express D1 receptors. Thus, increased suppression by dopamine of firing of VMN neurons that signal satiety with a rise in the discharge rate, and changed expression or activity of dopamine receptors might contribute to increased feeding behavior in juvenile rats hyperphagic and overweight due to early postnatal overfeeding. PMID- 11929196 TI - The effect of docosahexaenoic acid on aggression in elderly Thai subjects--a placebo-controlled double-blind study. AB - Docosahexaenoic acid (DHA) administration controls extraggression (aggression against others) in young subjects under psychological stress. However, it is not known whether its administration affects extraggression of elderly subjects. Forty Thai subjects of 50-60 years of age (22 males and 18 females) were recruited from Silpakorn University and nearby villages. They were allocated to the control and DHA groups in a double-blind fashion, and took 10 mixed plant oil capsules and 10 DHA capsules (1.5g DHA/day) for 2 months, respectively. Extraggression was measured with a psychological test (PF Study) at the beginning and end of the study. Just prior to the PF Study at the end of the study, subjects were asked to watch a stressful videotape as a stressor component. The average DHA intake from food was 150-160mg/day. In the group of university employees, extraggression did not change over time with placebo, whereas extraggression significantly decreased (31 +/- 13 to 24 +/- 13%, P = 0.04 by the paired-t test, P = 0.04 by ANOVA). In the group of villagers, there was no significant difference between the control and DHA groups in extraggression. The DHA administration favorably controlled extraggression in at least elderly white collar workers. The daily intake of 150-160 mg/day of DHA was not enough to control extraggression. PMID- 11929197 TI - Docosahexaenoic acid-rich phospholipid supplementation: effect on behavior, learning ability, and retinal function in control and n-3 polyunsaturated fatty acid deficient old mice. AB - This study investigated the effects of docosahexaenoic acid (DHA)-rich phospholipid supplementation on behavior, electroretinogram and phospholipid fatty acid (PUFA) composition in selected brain regions and retina in old mice. Two groups of mice were fed a semisynthetic balanced diet or a diet deficient in alpha-linolenic acid. At the age of 8 months, half of each diet group was supplemented with DHA. In the open field, no differences in motor or exploratory activities were observed between the four diet groups. In the light/dark test of anxiety, the time spent in the light compartment was significantly higher in both supplemented groups than in control and deficient groups. Learning performance in the Morris water maze was significantly impaired in deficient old mice, but was completely restored by the phospholipid supplementation. The electroretinogram showed a significant alteration of a- and b-wave amplitudes in control compared to deficient mice. Phospholipid supplementation induced a significant increase of b-wave amplitude in both control and deficient groups and restored normal fatty acid composition in brain regions and retina in deficient mice. DHA-rich phospholipids may improve learning ability, visual function and reverse biochemical modifications in old mice fed an n-3 polyunsaturated fatty acid deficient diet; they also may improve visual function in old mice fed a balanced diet. PMID- 11929198 TI - Perinatal undernutrition: changes in brain opiate receptor density. AB - The present work sought to study the binding properties of central mu-opiate receptors in whole brain and in different central areas in adult rats undernourished at perinatal age. Rats were undernourished with a hypoproteic diet containing 8% casein from day 14 of gestation until 50 days of age. The animals were thereafter fed a balanced commercial chow until 140 days of age. At this time point the experiments started. 3H-D-Ala2, N-Me-Phe4, Gly5-ol-enkephalin (3H DAMGO) was used to selectively label the mu-receptors. The results obtained demonstrated that perinatal undernutrition induced, in the adult animal, a decreased mu-receptors density (Bmax) both in whole brain as well as in midbrain, without significant changes in affinity. In addition, no changes were found in mu specific binding in the cortex of these undernourished animals. Taking into account that recent evidences from our laboratory have demonstrated a lower stress-induced analgesia following exposure to different stressful situations in rats undernourished in early life, the present findings seem to suggest that this lower analgesic response could be due, at least in part, to a lower density of mu opiate receptors in the brain. PMID- 11929199 TI - Zinc-deficient rats are insensitive to glucoprivation caused by 2-deoxy-D glucose. AB - Three-choice macronutrient intake studies indicate that zinc-deficient (Zn-) rats selectively decrease intake of carbohydrate. Because glucoprivic stimuli increase food intake and selection for carbohydrate, the ability of Zn- rats to respond to glucoprivation induced by 2-deoxy-D-glucose (2-DG) was tested. Rats were fed a Zn adequate (Zn+) or Zn- diet. In part 1, rats were challenged with 0, 250, or 400 mg 2-DG/kg BW (i.p.) after zinc deficiency was established. In part 2, rats received saline or 2-DG while zinc deficiency was being induced and then after deficiency was established. Food intake was increased after injection of 2-DG to Zn+ rats; however, food intake was not higher after 2-DG administration to Zn- rats. A dose-response test for 2-DG further confirmed these results. In part 2, it was found that Zn- rats lose the response to 2-DG administration when zinc deficiency-induced anorexia begins, after 3 days of consuming a zinc-deficient diet. It appears that the ability to sense blood glucose concentrations may be impaired during zinc deficiency, and this impairment could be a part of the anorexia that develops during zinc deficiency in the rat. PMID- 11929200 TI - Mediterranean diet and stroke: objectives and design of the SUN project. Seguimiento Universidad de Navarra. AB - PURPOSE: The Mediterranean diet has been postulated as a protective factor against different diseases including stroke. Thus, an epidemiological study in a Mediterranean country, such as Spain, focused on diet may offer new insights of the potential benefits of this nutritional pattern to prevent the onset of cerebrovascular diseases. METHODS: The SUN ("Seguimiento Universidad de Navarra") project is a prospective study among Spanish university alumni, aimed to identify the dietary determinants of stroke, coronary disease and other disorders. Two pilot studies have been developed. The first pilot study was focused on the understanding of the questionnaire. The second study used a random sample to assess the response proportion and the feasibility of using a mailing system for following-up the cohort. The first informative results are expected to be available after the first four years of following-up the cohort (2005). Here, we report the description of the baseline diet of the first participants in the cohort using data from 1,587 men and 2,260 women. RESULTS: The outcome of our pilot studies ensure the feasibility of a mail-based cohort. In the baseline assessment, we found a high consumption of olive oil (18.5 g/person/day), red wine (28.8 g/person/day), legumes (102.5 g/person/day), vegetables (507.8 g/person/ day) and fruits (316.7 g/person/day), with a great between-subject variability. Also, the values for cereals (170.4g/person/day), dairy products (239.3 g/person/day) and meat and meat products consumption (186 g/person/ day) in this cohort were estimated. The coefficients of variation ranged in women from 56 (for vegetables) to 240% (for red wine) and in men from 62 to 180% (for these same two items), reflecting a wide heterogeneity in the diet of participants. CONCLUSIONS: Although the participation was not high (22% according to the estimates of the pilot study), it was comparable to the proportion found in large previous cohorts such as the Nurses-II Health Study (24%). The sharp contrast in dietary habits between the US and Spain together with the high between-subjects variability we have found in our Spanish cohort provides an exceptional opportunity to assess the aspects of the Mediterranean diet, which may be protective against stroke and other neurological disorders. PMID- 11929201 TI - Anti-inflammatory medications: selective COX-2 inhibitors. PMID- 11929202 TI - Minimally invasive techniques for the treatment of intervertebral disk herniation. AB - Hemilaminectomy with diskectomy, the original surgical option to address intervertebral disk herniation, was superseded by open microdiskectomy, a less invasive technique recognized as the surgical benchmark with which minimally invasive spine surgery techniques have been compared as they have been developed. These minimally invasive surgical techniques for patients with herniated nucleus pulposus and radiculopathy include laser disk decompression, arthroscopic microdiskectomy, laparoscopic techniques, foraminal endoscopy, and microendoscopic diskectomy. Each has its own complications and requires a long learning curve to develop familiarity with the technique. Patient selection, and especially disk morphology, are the most important factors in choice of technique. The optimal candidate has a previously untreated single-level herniation with limited migration or sequestration of free fragments. PMID- 11929203 TI - Evaluation of pain in patients with apparently solidly fixed total hip arthroplasty components. AB - The cause of pain in a patient with an apparently solidly fixed total hip arthroplasty can be difficult to elucidate. A detailed history, careful examination, and plain radiographs provide the most useful information, especially in excluding causes not primarily related to the hip. Determining whether the pain is related to the implant, to soft tissue, or to bone can require laboratory tests, radiographic and fluoroscopic imaging, and contrast arthrography and local anesthetic injections. Particularly when pain is caused by occult infection, erythrocyte sedimentation rate, C-reactive protein level, hip aspiration, advanced radiologic imaging, and nuclear medicine scans can help determine the diagnosis. PMID- 11929204 TI - Physician-patient communication: a lost art? AB - In the face of rapid advances in technology, there has been a progressive deterioration of effective physician-patient communication. The American Academy of Orthopaedic Surgeons has identified that patients rate the orthopaedic profession as high in technical and low in communication skills. Poor communication, especially patient-interviewing skills, has been identified in medical students as well as in practicing physicians. Effective communication is associated with improved patient and physician satisfaction, better patient compliance, improved health outcomes, better-informed medical decisions, and reduced malpractice suits, and it likely contributes to reduced costs of care. Recognition of the importance of communication has influenced medical schools to revise curricula and to teach communication skills in residency training and continuing medical education programs. National certifying examinations also are being designed to incorporate these skills. Although written material is useful in increasing awareness of the importance of good physician-patient communication, behavioral change is more likely to occur in a workshop environment. The American Academy of Orthopaedic Surgeons is taking leadership in designing and implementing such an approach for its membership. PMID- 11929205 TI - Posttraumatic elbow stiffness: evaluation and management. AB - Posttraumatic elbow stiffness is a common problem that is often difficult to manage. The goal of treatment is to restore a functional range of elbow motion (> or =30 degrees to 130 degrees ). Nonsurgical treatment includes physical therapy and splinting. If nonsurgical treatment has failed, the type of surgical treatment required depends on the extent of degenerative changes. When degenerative changes are absent or mild, soft-tissue release offers reliable increases in elbow motion. When moderate degenerative changes exist within the joint, debridement arthroplasty of osteophytes and soft tissue has shown some success with increase in joint motion. With advanced degenerative changes, the therapeutic options are more limited. Results from biologic resurfacing arthroplasty are unpredictable, and total elbow arthroplasty should be reserved for the lower-demand elbow in a physiologically older individual. PMID- 11929206 TI - Pain management after major orthopaedic surgery: current strategies and new concepts. AB - Several recently developed analgesic techniques effectively control pain after major orthopaedic surgery. Neuraxial analgesia provided by epidural and spinal administration of local anesthetics and opioids provides the highest level of pain control; however, such therapy is highly invasive and labor intensive. Neuraxial analgesia is contraindicated in patients receiving low-molecular-weight heparin. Continuous plexus and peripheral neural blockades offer excellent analgesia without the side effects associated with neuraxial and parenteral opioids. Intravenous patient-controlled analgesia allows patients to titrate analgesics in amounts proportional to perceived pain stimulus and provide improved analgesic uniformity. Oral sustained-release opioids offer superior pain control and greater convenience than short-duration agents provide. Opioid dose requirements may be reduced by coadministration of COX-2-type nonsteroidal analgesics. PMID- 11929207 TI - Saphenous neuritis: a poorly understood cause of medial knee pain. AB - Saphenous neuritis is a painful condition caused by either irritation or compression at the adductor canal or elsewhere along the course of the saphenous nerve. The condition also may be associated with surgical or nonsurgical trauma to the nerve, especially at the medial or anterior aspect of the knee. Saphenous neuritis can imitate other pathology around the knee, particularly a medial meniscal tear or osteoarthritis. Unrecognized saphenous neuritis can confuse the patient's clinical picture, complicate treatment, and compromise results. As an isolated entity, saphenous neuritis may appear in conjunction with other common problems, such as osteoarthritis and patellofemoral pain syndrome, and it can have an indolent and protracted course. Its clinical appearance is characterized by allodynia along the course of the saphenous nerve. The diagnosis is confirmed by relief of symptoms after injection of the affected area with local anesthetic. Initial treatment can include non-surgical symptomatic care, treatment of associated pathology, and diagnostic or therapeutic injections of local anesthetic. In recalcitrant cases, surgical decompression and neurectomy are potential options. The key to treatment is prompt recognition; palpation of the saphenous nerve should be part of every routine examination of the knee. PMID- 11929208 TI - Duchenne muscular dystrophy. AB - Duchenne muscular dystrophy is an X-linked disease of muscle caused by an absence of the protein dystrophin. Affected boys begin manifesting signs of disease early in life, cease walking at the beginning of the second decade, and usually die by age 20 years. Until treatment of the basic genetic defect is available, medical, surgical, and rehabilitative approaches can be used to maintain patient function and comfort. Corticosteroids, including prednisone and a related compound, deflazacort, have recently been shown to markedly delay the loss of muscle strength and function in boys with Duchenne muscular dystrophy. Surgical release of lower extremity contractures may benefit some patients. Approximately 90% of boys with Duchenne muscular dystrophy will develop severe scoliosis, which is not amenable to control by nonsurgical means such as bracing or adaptive seating. The most effective treatment for severe scoliosis is prevention by intervening with early spinal fusion utilizing segmental instrumentation as soon as curves are ascertained and before the onset of severe pulmonary or cardiac dysfunction. PMID- 11929209 TI - The DNA damage response in filamentous fungi. AB - The mechanisms used by fungal cells to repair DNA damage have been subjects of intensive investigation for almost 50 years. As a result, the model yeasts Schizosaccharomyces pombe and Saccharomyces cerevisiae have led the way in yielding critical insights into the nature of the DNA damage response. At the same time, largely through the efforts of Etta Kafer, Hirokazu Inoue, and colleagues, a substantial collection of Aspergillus nidulans and Neurospora crassa DNA repair mutants has been identified and characterized in detail. As the analysis of these mutants continues and increasing amounts of annotated genome sequence become available, it is becoming readily apparent that the DNA damage response of filamentous fungi possesses several features that distinguish it from the model yeasts. These features are emphasized in this review, which describes the genes, regulatory networks, and processes that compose the fungal DNA damage response. Further characterization of this response will likely yield general insights that are applicable to animals and plants. Moreover, it may also become evident that the DNA damage response can be manipulated to control fungal growth. PMID- 11929210 TI - Non-Mendelian inheritance revealed in a genetic analysis of sexual progeny of Phytophthora cinnamomi with microsatellite markers. AB - We report the development of four microsatellite loci into genetic markers for the diploid oomycete plant pathogen Phytophthora cinnamomi and that (AC)(n) and (AG)(n) microsatellites are significantly less frequent than in plant and mammal genomes. A minisatellite motif 14 bp long was also discovered. The four microsatellite loci were used to analyze sexual progeny from four separate crosses of P. cinnamomi. A large proportion of non-Mendelian inheritance was observed across all loci in all four crosses, including inheritance of more than two alleles at a locus and noninheritance of alleles from either parent at a locus. The aberrant inheritance is best explained by nondisjunction at meiosis in both the A1 parent and the A2 trisomic parents, resulting in aneuploid progeny. Two loci on the putative trisomic chromosome showed linkage and no loci were linked to mating type. One aneuploid offspring was shown to have lost alleles at two loci following subculture over 4 years, indicating that aneuploid progeny may not be mitotically stable. PMID- 11929211 TI - Role in pathogenesis of two endo-beta-1,4-xylanase genes from the vascular wilt fungus Fusarium oxysporum. AB - A gene, xyl4, whose predicted amino acid sequence shows significant homology with family 11 xylanases, was identified from the tomato vascular wilt fungus Fusarium oxysporum f. sp. lycopersici. Expression of xyl4 is induced on oat spelt xylan as the carbon source, subject to carbon catabolite repression and preferentially expressed at alkaline ambient pH. Transcript levels of xyl4 on an inducing carbon source are differentially regulated by the nature and concentration of the nitrogen source. As shown by RT-PCR, xyl4 is expressed by F. oxysporum during the entire cycle of infection on tomato plants. Targeted inactivation of xyl4 and of xyl3, a previously identified gene of F. oxysporum f. sp. lycopersici encoding a family 10 xylanase, had no detectable effect on virulence on tomato plants, demonstrating that both genes are not essential for pathogenicity. PMID- 11929212 TI - Specific and reversible inactivation of Phycomyces blakesleeanus isocitrate lyase by ascorbate-iron: role of two redox-active cysteines. AB - Phycomyces blakesleeanus isocitrate lyase (EC 4.1.3.1) is in vivo reversibly inactivated by hydrogen peroxide. The purified enzyme showed reversible inactivation by an ascorbate plus Fe(2+) system under aerobic conditions. Inactivation requires hydrogen peroxide; was prevented by catalase, EDTA, Mg(2+), isocitrate, GSH, DTT, or cysteine; and was reversed by thiols. The ascorbate served as a source of hydrogen peroxide and also reduced the Fe(3+) ions produced in a "site-specific" Fenton reaction. Two redox-active cysteine residues per enzyme subunit are targets of oxidative modification; one of them is located at the catalytic site and the other at the metal regulatory site. The oxidized enzyme showed covalent and conformational changes that led to inactivation, decreased thermal stability, and also increased inactivation by trypsin. These results represent an example of redox regulation of an enzymatic activity, which may play a role as a sensor of redox cellular status. PMID- 11929213 TI - A genetic map of Blumeria graminis based on functional genes, avirulence genes, and molecular markers. AB - A genetic map of the powdery mildew fungus, Blumeria graminis f. sp. hordei, an obligate biotrophic pathogen of barley, is presented. The linkage analysis was conducted on 81 segregating haploid progeny isolates from a cross between 2 isolates differing in seven avirulence genes. A total of 359 loci were mapped, comprising 182 amplified fragment length polymorphism markers, 168 restriction fragment length polymorphism markers including 42 LTR-retrotransposon loci and 99 expressed sequence tags (ESTs), all the seven avirulence genes, and a marker closely linked to the mating type gene. The markers are distributed over 34 linkage groups covering a total of 2114 cM. Five avirulence genes were found to be linked and mapped in clusters of three and two, and two were unlinked. The Avr(a6) gene was found to be closely linked to markers suitable for a map-based cloning approach. A linkage between ESTs allowed us to demonstrate examples of synteny between genes in B. graminis and Neurospora crassa. PMID- 11929214 TI - Hydrophobins DGH1, DGH2, and DGH3 in the lichen-forming basidiomycete Dictyonema glabratum. AB - Dictyonema glabratum is a lichen-forming basidiomycete whose symbiotic phenotype shares similarities to both lichens and its non-lichen-forming relatives. In the photobiont layer of D. glabratum intercellular gas-filled spaces are present even when the lichen is water-saturated. The walls of hyphae lining air cavities are covered by a hydrophobic, rodlet-patterned layer, assumed to be formed by hydrophobins. Hot SDS-insoluble, but trifluoroacetic acid-soluble lichen cell wall extracts contained seven proteins. The N-terminal sequence of the most abundant 14-kDa protein was used to carry out cDNA cloning by RT-PCR. The deduced amino acid sequence of the amplified fragment encoded a class I hydrophobin, called DGH1. The cDNA sequence encoding the signal peptide was cloned by RACE PCR, which also coamplified cDNA fragments encoding two additional class I hydrophobins, DGH2 and DGH3. The three proteins share 54 to 66% amino acid identity. The D. glabratum hydrophobin extract containing either all proteins or primarily DGH1 self-assembled and formed a rodlet mosaic similar to the one observed in situ. Concentration of the protein extract was shown to influence the length of the self-assembled rodlets. PMID- 11929216 TI - Isolation and characterization of the mating-type idiomorphs from the wheat septoria leaf blotch fungus Mycosphaerella graminicola. AB - Both mating-type loci from the wheat septoria leaf blotch pathogen Mycosphaerella graminicola have been cloned and sequenced. The MAT1-2 gene was identified by screening a genomic library from the MAT1-2 isolate IPO94269 with a heterologous probe from Tapesia yallundae. The MAT1-2 idiomorph is 2772 bp and contains a single gene encoding a putative high-mobility-group protein of 394 amino acids. The opposite idiomorph was obtained from isolate IPO323, which has the complementary mating type, by long-range PCR using primers derived from sequences flanking the MAT1-2 idiomorph. The MAT1-1 locus is 2839 bp in size and contains a single open reading frame encoding a putative alpha1-domain protein of 297 amino acids. Within the nonidiomorphic sequences, homology was found with palI, encoding a membrane receptor from Aspergillus nidulans, and a gene encoding a putative component of the anaphase-promoting complex from Schizosaccharomyces pombe and a DNA-(apurinic or apyrimidinic) lyase from S. pombe. For each of the MAT genes specific primers were designed and tested on an F1 mapping population that was generated from a cross between IPO323 and IPO94269. An absolute correlation was found between the amplified allele-specific fragments and the mating type as determined by backcrosses of each F1 progeny isolate to the parental isolates. The primers were also used to screen a collection of field isolates in a multiplex PCR. An equal distribution of MAT1-1 and MAT1-2 alleles was found for most geographic origins examined. PMID- 11929215 TI - Molecular cloning, characterization, and expression of a redox-responsive cutinase from Monilinia fructicola (Wint.) Honey. AB - cDNA clones encoding a cutinase expressed in cutin-induced cultures of the plant pathogen Monilinia fructicola were isolated using a protein-based strategy. The largest cDNA (Mfcut1) was found to contain an open reading frame of 603 bp that predicted a 20.2-kDa protein of 201 amino acids with a 20-amino-acid secretory signal peptide and a pI of 8.4. The predicted protein contained cutinase/lipase consensus sequences with active site serines and potential protein kinase phosphorylation sites. Comparison of the deduced amino sequence from Mfcut1 with other fungal cutinase sequences revealed new features, which include conserved cysteines, C-terminal aromatic residues, and a novel histidine substitution in the D-H active site motif. The presence in the growth medium of antioxidants, such as caffeic acid, suppressed mRNA accumulation and enzyme activity of a cutinase from M. fructicola. MFCUT1 was expressed at high levels as a His-tagged fusion protein in Pichia pastoris and purified to apparent homogeneity in a single step by Ni(2+)-nitrilotriacetic acid affinity chromatography. Analysis of variant MFCUT1 mutants in which the novel serine and histidine residues were replaced by site-directed mutagenesis indicated that these residues had an important effect on enzyme activity. PMID- 11929217 TI - The role of proline in osmoregulation in Phytophthora nicotianae. AB - A cDNA encoding Delta(1)-pyrroline-5-carboxylate reductase (P5CR), the enzyme that catalyzes the final step in proline biosynthesis in plants and bacteria, has been cloned from the oomycete plant pathogen Phytophthora nicotianae. Genomic DNA blots indicated that P. nicotianae and P. cinnamomi each contain a single P5CR gene, whereas P. infestans contains one or two genes. Complementation of a strain of Escherichia coli defective in the P5CR protein by the P. nicotianae P5CR cDNA confirmed that the gene encoded a functional P5CR. RNA blots revealed that P5CR was expressed at a much higher level in P. nicotianae zoospores than in vegetative hyphae, sporulating hyphae, or germlings. Furthermore, P5CR mRNA levels increased with time in zoospores, demonstrating that transcription occurs in zoospores. mRNA encoding histidine and tryptophan biosynthetic enzymes was not highly and specifically expressed in zoospores, indicating that the developmental pattern of P5CR expression was not simply a reflection of overall amino acid biosynthesis as might be required for protein synthesis. Measurement of free proline in P. nicotianae at different stages of the life cycle revealed that proline concentration was highest in sporulating hyphae. Vegetative hyphae and germlings contained about 50% of this concentration of proline, and zoospores contained only about 1% of this level. Substantial amounts of proline were measured in the medium into which the zoospore had been released. Hypoosmotic shock of P. nicotianae hyphae led to an approximately 50% decrease in free proline concentration within 30 min of transfer to low-osmolarity medium and was accompanied by an increase in the level of P5CR mRNA. These data are discussed in terms of the possible role of proline in osmoregulation in Phytophthora. PMID- 11929218 TI - Hydrogen-bond-mediated tuning of the redox potential of the non-heme Fe site of superoxide dismutase. AB - The highly homologous proteins of Fe-containing superoxide dismutase (FeSOD) and MnSOD from Escherichia coli nonetheless exert very different redox tuning on the active site metal ion [Vance; Miller J. Am. Chem. Soc. 1998, 120, 461-467; Biochemistry 2001, 40, 13079-13087]. This was proposed to stem from different hydrogen bonding between the protein and the metal ion's coordinated solvent molecule, and the tight coupling between the protonation state of coordinated solvent and the oxidation state of the metal ion. We now present density functional theory (DFT) calculations on Fe2+ and Fe3+ bound to models of both FeSOD and MnSOD. The calculations support a very important role for the conserved second sphere Gln in MnSOD in specifically destabilizing coordinated H2O relative to coordinated OH-, and thus disfavouring the oxidized state of the metal ion. To test these results we have mutated this Gln to Glu, which is isosteric and isoelectronic to Gln but functions as an H-bond acceptor instead of an H-bond donor and thus should increase the stability of Fe2+-bound H2O. In accordance with the calculations, Q69E-FeSOD displays a significantly higher reduction potential than wild-type FeSOD. Thus we have demonstrated that hydrogen bonds to coordinated solvent can exert strong redox tuning on a metal ion. PMID- 11929219 TI - A fast and easy computational method to calculate the (13)C NMR chemical shift of organic species adsorbed on the zeolite surface. AB - Calculations of the 13C NMR chemical shifts for the methoxy and ethoxy groups adsorbed on Y and ZSM-5 zeolites were computed at GIAO/B3LYP/6-31+G*//MM+ level of theory, using a cluster representing a real part of the zeolites. The Y zeolite was represented by a cluster with 168 atoms, while ZSM-5 was represented by a cluster with 144 atoms. The calculated chemical shifts agreed well with reported experimental values, showing that the difference in chemical shifts is associated with differences in the geometry of the alkoxides on the two zeolites. PMID- 11929220 TI - Heterosuperbenzenes: a new family of nitrogen-functionalized, graphitic molecules. AB - A versatile synthetic route has been demonstrated to provide a series of soluble, fused polyaromatic C,N-molecules, with tunable optical and electronic properties and ligand functionality. The N-functionalized heterosuperbenzene, 1, is the founding member of this new family of active graphitic substructures. PMID- 11929221 TI - Low-activation solid-state syntheses by reducing transport lengths to atomic scales as demonstrated by case studies on AgNO(3) and AgO. AB - We have studied solid-state reactions of educt mixtures of elements in an atomic dispersion. The reduced transport distances allow for extremely low activated reactions. This has been demonstrated by case studies on AgNO3 and AgO, which form and crystallize from the atoms below room temperature. PMID- 11929222 TI - The role of configurational entropy in biochemical cooperativity. AB - Cooperativity is a common biochemical phenomenon in which two or more otherwise independent processes are thermodynamically coupled. Because cooperative processes are usually attended by changes in molecular conformation, thermodynamic coupling is usually attributed to an enthalpy-driven mechanism. In the family of glycopeptide antibiotics that includes vancomycin, however, cooperative phenomena occur that cannot be explained by conformational change. In this communication, we demonstrate that cooperativity in these systems can arise solely from changes in vibrational activity. PMID- 11929223 TI - Total synthesis and proof of structure of mycothiol bimane. AB - Mycothiol is a low-molecular weight thiol produced by actinomycetes that serves to protect these organisms from oxidative stress and alkylating agents. We report the total synthesis of mycothiol bimane (1) which is a commonly isolated derivative of mycothiol. The synthesis confirms the original structure assignment and unambiguously establishes the absolute stereochemistry of mycothiol to be 1-d myo-inosityl 2-deoxy-2-(N-acetamido-l-cysteinamido)-alpha-d-glucopyranoside. PMID- 11929224 TI - Crystallographic characterization and structural analysis of the first organic functionalization product of the endohedral fullerene Sc(3)N@C(80). AB - The structure of Sc3N@C80-C10H12O2, a Diels-Alder cycloadduct of Sc3N@C80, has been determined. The crystallographic data shows that cycloaddition occurs at a C C bond of 6:5 ring junction, and that the fullerene C1-C2 bond is elongated and pulled out from the fullerene. The Sc3N unit is well-ordered within the C80 cage and positioned away from the site of addition. The proximity of the Sc atoms to the cage carbon atoms causes those carbon atoms to protrude slightly from the surface of the fullerene cage. PMID- 11929225 TI - Epoxyquinol A, a highly functionalized pentaketide dimer with antiangiogenic activity isolated from fungal metabolites. AB - A unique pentaketide dimer structure of a novel fungal metabolite with antiangiogenic activity, designated as epoxyquinol A (1), was determined on the basis of NMR spectral data as well as the X-ray crystallographic analysis. 1 inhibits the endothelial migration induced by vascular endothelial growth factor (ED100 = 3 mug/mL). PMID- 11929226 TI - Enzymatic synthesis of chiral organophosphothioates from prochiral precursors. AB - The phosphotriesterase from Pseudomonas diminuta has been shown to selectively cleave the pro-R p-nitrophenolate substituent from bis-p-nitrophenyl alkyl phosphothioate esters. When the alkyl substituent is methyl, ethyl, or isopropyl the enantiomeric excess of the product is >/=99%. Manipulation of the active site through mutagenesis has enabled the preparation of protein variants that preferentially hydrolyze the pro-S substituent of the target substrates. This methodology thus permits the preparation of chiral products from prochiral precursors. PMID- 11929228 TI - Stereoselective cyclopropanation by cyclocopolymerization of butadiene. AB - An unprecedented cyclopropanation by cyclopolymerization reaction is presented. In particular, catalytic copolymerization processes of ethene and butadiene lead with high trans selectivity to a complete cyclocopolymerization of butadiene units. Ethene-based copolymers including cyclopropane and cyclopentane rings, which can be relevant to polyolefin applications, are obtained with high yields. Several aspects relative to the mechanism of this cyclocopolymerization reaction, including the high stereoselectivity, are discussed. PMID- 11929227 TI - Nickel-BINAP catalyzed enantioselective alpha-arylation of alpha-substituted gamma-butyrolactones. AB - A Ni(0)-BINAP system is utilized for the highly enantioselective alpha-arylation of alpha-substituted gamma-butyrolactones with aryl chlorides and bromides. alpha Quaternization is achieved in moderate to excellent yields. Furthermore, the rate accelerating effect caused by the addition of Zn(II) salts is investigated. PMID- 11929229 TI - Photoinduced supramolecular chirality in amorphous azobenzene polymer films. AB - Supramolecular chirality was optically induced in amorphous and achiral azobenzene polymer films by irradiation of a laser beam with elliptical polarization. The chirality resulted from helical orientation of azobenzene chromophores by a combined process of circular and linear polarization. The helix handedness could be controlled by incident light-handedness. PMID- 11929230 TI - Ruthenium-mediated cycloaromatization of acyclic enediynes and dienynes at ambient temperature. AB - The ruthenium(II) cation, [Cp*Ru(NCMe)3]OTf (4), triggers the Bergman cycloaromatization of acyclic endiynes at room temperature in THF solvent. Treatment of 1,2-di(1-alkynynyl)cyclopentenes (13-Me, alkynyl = propynyl; 13-Prn, alkynyl = pentynyl; 13-Bui, alkynyl = 4-methyl-pent-1-ynyl) with 4 in THF solvent at room temperature gives rise to the ruthenium arene complexes: [Cp*Ru{(3a,4,5,6,7,7a-eta)-2,3-dihydro-5,6-dialkyl-1H-indene}]OTf (15-Me, alkyl = methyl, 64% yield; 15-Prn, alkyl = n-propyl, 73% yield; 15-Bui, alkyl = 4-methyl 1-pentynyl, 88% yield). In a similar fashion, the room-temperature reaction of 4 with 1-ethynyl-2-(1-propynyl)cyclopentene (11) and [2-(1-propynyl)-1-cyclopenten 1-yl]trimethylsilane (14) leads to the formation of [Cp*Ru{(3a,4,5,6,7,7a-eta) 2,3-dihydro-5-methyl-1H-indene}]OTf (12, 92% yield) and [Cp*Ru{(3a,4,5,6,7,7a eta)-2,3-dihydro-6-methyl-1H-inden-5-yl)trimethylsilane}]OTf (16, 77% yield), respectively. The bis(TMS)-substituted enediyne (1-cyclopentene-1,2-diyldi-2,1 ethynediyl)bis(trimethylsilane) (9-TMS) and 4 underwent reaction at 100 degrees C to give [Cp*Ru{(3a,4,5,6,7,7a-eta)-2,3-dihydro-1H-inden-5-yl)trimethylsilane}]OTf (10, 69% yield). Deuterium-labeling studies rule out a mechanism that involves a ruthenium-vinylidene intermediate, and provide support for the involvement of a p benzyne intermediate. In a similar fashion, complex 4 is shown to trigger the cycloaromatization of the conjugated dienyne, 1-ethenyl-2-(1 pentynyl)cyclopentene (19), at room temperature in chloroform-d1 solvent to give [Cp*Ru{(3a,4,5,6,7,7a-eta)-2,3-dihydro-5-(1-propyl)-1H-indene}]OTf (20, 96% yield), with no deuterium enrichment. In the absence of ruthenium the thermal cyclization reactions of unsubstituted acyclic enediynes (Bergman cycloaromatization) and acyclic conjugated dienynes (Hopf cyclization) typically require elevated temperatures (150-250 degrees C). Complexes 10 and 15-Prn were characterized structurally by X-ray crystallography. PMID- 11929231 TI - Selective binding of mannose-encapsulated gold nanoparticles to type 1 pili in Escherichia coli. AB - The synthesis, characterization and biological application of mannose encapsulated gold nanoparticles (m-AuNP) are reported. m-AuNP is well dispersed and very stable without aggregation in the media of broad ion strength and pH ranges. The selective binding of m-AuNP to the mannose adhesin FimH of bacterial type 1 pili is demonstrated using transmission electron microscopy. The competition assay with free mannose suggests that m-AuNP binds FimH better than free mannose does. This work demonstrates that carbohydrate attached nanoparticles can be used as an efficient affinity label and a multi-ligand carrier in a biological system. PMID- 11929232 TI - A modular assembly strategy for improving the substrate specificity of small catalytic peptides. AB - In contrast to large proteins, small peptide catalysts typically display limited specificity for small molecule substrates. This is presumably a result of the limited opportunities small peptides have to fold in a manner that provides for the formation of an isolated reaction vessel that effectively binds and sequesters substrates from bulk solvent while at the same time catalyzing their transformation. For the preparation of small peptide catalysts that possess improved substrate specificity, we have developed a modular assembly strategy that involves appending phage display-derived substrate binding-domain modules to catalytically active peptide domains. We demonstrate the potential of this strategy with the construction of a small 35-amino acid residue aldolase peptide with improved substrate specificity. The advantages of this approach are that it reduces the demand on the functionalization of the catalytic site and it is modular, therefore making its adaptation to a variety of specificities rapid. The modular assembly strategy studied here may present advantages over exhaustive searches of large random-sequence peptide libraries for peptides with singular function. PMID- 11929233 TI - Tetracyanide-bridged divanadium complexes: redox switching between strong antiferromagnetic and strong ferromagnetic coupling. AB - Reaction of [(Me3tacn)V(CF3SO3)3] (Me3tacn = N,N',N''-trimethyl-1,4,7 triazacyclononane) with LiCN in DMF results in oligomerization of cyanide to form [(Me3tacn)2V2(CN)4(mu-C4N4)]. The structure of this binuclear complex features a planar tetracyanide unit bridging two VIV centers via imido type linkages. The conjugated pathway provided by the bridging ligand leads to strong antiferromagnetic coupling (J = -112 cm-1) and an S = 0 ground state. Reduction of the complex with cobaltocene generates the Class III mixed-valence anion [(Me3tacn)2V2(CN)4(mu-C4N4)]1-, wherein resonance exchange induces strong ferromagnetic coupling to give a well-isolated S = 3/2 ground state. PMID- 11929235 TI - Negative differential resistance in phenylene ethynylene oligomers. AB - The origin of the sharp peak profile (i.e., negative differential resistance, NDR) observed in the I/V curves of three-ring phenylene ethynylene oligomers is a topic of major current interest. Here, quantum-chemical calculations are performed to analyze the evolution of the one-electron structure of an unsubstituted three-ring oligomer under the influence of a static electric field (which models the driving voltage applied in the experiments). The results indicate that the rotation of the central ring of the oligomer induces resonant tunneling processes over a limited voltage range. This can thus be responsible for the NDR signature observed experimentally. PMID- 11929236 TI - Selective syntheses of metalated pyridines from two different unsymmetrical acetylenes, a nitrile, and a titanium(II) alkoxide. AB - Cyclotrimerization of two different, unsymmetrical acetylenes and p toluenesulfonylnitrile with a divalent titanium alkoxide reagent, Ti(O-i-Pr)4/2 i PrMgCl, yielded single pyridyltitanium compounds in a highly selective manner. These metalated pyridines were confirmed by deuteriolysis to give the corresponding deuterated pyridines and underwent iodinolysis and copper-catalyzed alkylation to demonstrate their synthetic utility. Alternatively, a different type of cyclotrimerization of an alkynamide, terminal acetylenes, and alpha alkoxynitriles mediated by the same titanium(II) alkoxide again proceeded in a highly selective manner to give single pyridines having a titanated side chain. PMID- 11929234 TI - Magnetic resonance imaging detects a specific peptide-protein binding event. AB - DOTA was conjugated to the N-terminus of a 12-mer peptide by using standard peptide synthesis chemistry. The peptide, first isolated by phage display, maintained a high affinity for its protein-binding target, Gal-80, even with GdDOTA attached. The high affinity constant (KA = 5 x 105 M-1) combined with the high relaxivity of the resulting GdDOTA-peptide.protein complex (r1bound = 44.8 +/- 1.7 mM-1 s-1) allowed detection of Gal-80 at muM levels using a standard magnetic resonance imaging protocol. This novel peptide-based, binding-activated MRI method could potentially be used to screen a wide variety of biomolecules. PMID- 11929237 TI - Crystals of crystals: fabrication of encapsulated and ordered two-dimensional arrays of microcrystals. AB - A new technique-crystallization in asymmetric microwells-generates arrays of small crystals with controlled size, orientation, and arrangement in space. These arrays of crystals can be generated in a form completely encapsulated in polymer. PMID- 11929238 TI - Structural chemistry of a green fluorescent protein Zn biosensor. AB - We designed a green fluorescent protein mutant (BFPms1) that preferentially binds Zn(II) (enhancing fluorescence intensity) and Cu(II) (quenching fluorescence) directly to a chromophore ligand that resembles a dipyrrole unit of a porphyrin. Crystallographic structure determination of apo, Zn(II)-bound, and Cu(II)-bound BFPms1 to better than 1.5 A resolution allowed us to refine metal centers without geometric restraints, to calculate experimental standard uncertainty errors for bond lengths and angles, and to model thermal displacement parameters anisotropically. The BFPms1 Zn(II) site (KD = 50 muM) displays distorted trigonal bipyrimidal geometry, with Zn(II) binding to Glu222, to a water molecule, and tridentate to the chromophore ligand. In contrast, the BFPms1 Cu(II) site (KD = 24 muM) exhibits square planar geometry similar to metalated porphyrins, with Cu(II) binding to the chromophore chelate and Glu222. The apo structure reveals a large electropositive region near the designed metal insertion channel, suggesting a basis for the measured metal cation binding kinetics. The preorganized tridentate ligand is accommodated in both coordination geometries by a 0.4 A difference between the Zn and Cu positions and by distinct rearrangements of Glu222. The highly accurate metal ligand bond lengths reveal different protonation states for the same oxygen bound to Zn vs Cu, with implications for the observed metal ion specificity. Crystallographic anisotropic thermal factor analysis validates metal ion rigidification of the chromophore in enhancement of fluorescence intensity upon Zn(II) binding. Thus, our high-resolution structures reveal how structure-based design has effectively linked selective metal binding to changes in fluorescent properties. Furthermore, this protein Zn(II) biosensor provides a prototype suitable for further optimization by directed evolution to generate metalloprotein variants with desirable physical or biochemical properties. PMID- 11929239 TI - Sequestering of Eu(III) by a GAAA RNA tetraloop. AB - The site-specific binding of metal ions maintains an important role in the structure, thermal stability, and function of folded RNA structures. RNA tetraloops of the "GNRA" family (where N = any base and R = any purine), which owe their unusual stability to base stacking and an extensive hydrogen bonding network, have been observed to bind metal ions having different chemical and geometric properties. We have used laser-induced lanthanide luminescence and isothermal titration calorimetry (ITC) to examine the metal-binding properties of an RNA stem loop of the GNRA family. Previous research has shown that a single Eu(III) ion binds the stem loop fragment in a highly dehydrated site with a K(d) of approximately 12 microM. Curve-fitting analysis of the broad luminescence excitation spectrum of Eu(III) upon complexation with the tetraloop fragment indicates the possibility of two microenvironments that do not differ in hydration number. Binding of Eu(III) to the loop was accompanied by positive enthalpic changes, consistent with energetic cost of removal of water molecules and suggesting that the binding is entropically driven. By comparison, binding of Mg(II) or Mn(II) to the RNA loop, or Eu(III) to the DNA analogue of the loop, was associated with exothermic changes, consistent with predominantly outer-sphere coordination. These results suggest specific binding, most probably involving ligands on the 5' side of the loop. PMID- 11929240 TI - Kinetics of disassembly of a DNA-bound porphyrin supramolecular array. AB - trans-Bis(N-methylpyridinium-4-yl)diphenylporphine forms extended, organized assemblies on DNA templates under appropriate conditions of concentration, ionic strength, and temperature. Addition of beta-cyclodextrin to these arrays leads to their disassembly as evidenced by changes in extinction, circular dichroism, and resonance light scattering spectra. The structure or flexibility of the polymer template has an effect on the rate of disassembly; the reaction is faster on a poly(dG-dC)(2) surface than on ct DNA. The kinetic profiles for the disassembly process can be fit with great precision with a two-kinetic parameter equation in which the rate constant is itself a function of time. The reaction rate, studied in the presence of excess beta-CD, shows a dependence on the mode of detection. A model is presented to account for these observations in which the arrays become increasingly reactive with time due to beta-CD attack at the interior of the porphyrin assemblies as well as the ends. PMID- 11929241 TI - Internally mismatched RNA: pH and solvent dependence of the thermal unfolding of tRNA(Ala) acceptor stem microhairpins. AB - The thermal unfolding of two RNA hairpin systems derived from the aminoacyl accepting arm of Escherichia coli tRNA(Ala) that included all possible single internal mismatches mostly in the third base pair position was measured spectroscopically in 0.1 M NaCl at pH 7.5 and, in part, 5.5. The thermodynamic parameters DeltaH(o), DeltaS(o), DeltaG(o), and T(m) of a total of 36 RNA strands were determined through nonlinear curve fitting of the melting profiles (22 tetralooped 22mers and 14 heptalooped 25mers, same stem sequence). Only three of the 22mers, the A.C-containing variants, were shown to be significantly more stable at pH 5.5. A number of remarkable differences-most likely of more general relevance-between the thermodynamics of certain structurally very similar hairpin variants (e.g., G.C versus A.U, G.U versus I.U) at pH 7.5 are discussed with respect to two possible ways of helix stabilization: pronounced hydration versus low entropic penalty. Four selected 22mers were additionally analyzed in 1 M NaCl and in solvent mixtures containing ethanol, ethylene glycol, and dimethylformamide. The wealth of thermodynamic data suggest that the exothermicity DeltaH(o) and entropic penalty T x DeltaS(o) of folding are strongly dominated by the rearrangement and formation of hydration layers around the solutes, while it is well-known that the stability of folding results only from the difference (DeltaG(o)) and ratio of both parameters (T(m) = DeltaH (o)/DeltaS(o)). PMID- 11929242 TI - Thermodynamic correlation analysis: hydration and perturbation sensitivity of RNA secondary structures. AB - The thermodynamic parameters DeltaH, DeltaS, T(m), and DeltaG of a total of 36 RNA strands, 22 tetralooped 22mers, and 14 heptalooped 25mers (same stem sequence) were analyzed with respect to enthalpy-entropy compensation (EEC). The EEC plots [DeltaH, DeltaS] were compared with collected literature data from protein and nucleic acid unfolding studies (3224 and 241 datapoints, respectively) which all proved to be remarkably linear. The similarity of the compensation slopes and intercepts for all compounds indicate that, irrespective of the chemical nature and stability of the folding solutes, the exothermicity DeltaH and entropic penalty T x DeltaS of folding are strongly dominated by the rearrangement and formation of hydration layers around the solutes, while it is well-known that the stability of folding results only from the difference (DeltaG) and ratio (T(m)) of both parameters. EEC plots [DeltaH, DeltaS] are presented in an extended context, as 3D plots [DeltaH, DeltaS, T(m)] allowing for a correct analytical description of the enthalpy-entropy relationship and for more practical interpretations of large amounts of thermodynamic data when replotted as [DeltaH, T(m)] or [DeltaG(T), T(m)]. The introduction of a variety of mismatches into nucleic acids, or limited irreguliarities into any supramolecular complex, and the analysis of the involved thermodynamics as shown in this study-i.e., scanning the "enthalpy-entropy space" of whole macromolecular subgroups-should permit to extract and quantify more "hidden information", such as hydration extent and sensitivity of macromolecular frameworks toward desolvation and structural perturbation, from thermodynamic analyses of large sample sizes. PMID- 11929243 TI - Deoxyribozyme-based logic gates. AB - We report herein a set of deoxyribozyme-based logic gates capable of generating any Boolean function. We construct basic NOT and AND gates, followed by the more complex XOR gate. These gates were constructed through a modular design that combines molecular beacon stem-loops with hammerhead-type deoxyribozymes. Importantly, as the gates have oligonucleotides as both inputs and output, they open the possibility of communication between various computation elements in solution. The operation of these gates is conveniently connected to a fluorescent readout. PMID- 11929244 TI - Convergent synthesis of polycyclic ethers via the intramolecular allylation of alpha-acetoxy ethers and subsequent ring-closing metathesis. AB - The Lewis acid mediated reaction of alpha-acetoxy ethers 15-22 gave the corresponding cyclized products 23, 25, 27, 29, 31, 32, 34, and 36 in good yields with high stereoselectivities. Those cyclized products were subjected to ring closing metathesis to afford the polycyclic ethers 38-42, 44, and 45 in good yields. The usefulness of the present methodology was demonstrated by the convergent synthesis of the CDEF ring system of brevetoxin B (1) and the CDEFG ring system of gambierol (2). PMID- 11929245 TI - Reactions of N-methyl-N-(4-biphenylyl)nitrenium ion with electron-rich arenes: laser flash photolysis and product studies. AB - An arylnitrenium ion, N-methyl-N-(4-biphenylyl)nitrenium ion, was generated through photolysis of 1-(N-methyl-N-4-biphenylyl)amino-2,4,6-trimethylpyridinium tetrafluoroborate, and its reactions with various donor-substituted arenes (e.g., 1,3,5-trimethoxybenzene, mesitylene, 1,4-dimethoxybenzene, hexamethylbenzene, etc.) were examined using product analysis and laser flash photolysis. In general, trapping of the short-lived nitrenium ion by the arenes leads to three types of products: (1) the parent amine, N-methyl-N-4-biphenylylamine; (2) an ortho-adduct, where the ring position ortho to the nitrenium ion center is bonded to the arene ring; and (3) an N-adduct, where the nitrenium ion nitrogen is bonded to the trap. Laser flash photolysis studies show that the rates of these trapping reactions vary from 10(4) to 10(9) M(-1) s(-1), depending on the structure of the arene trap. These trapping rate constants do not correlate with the one-electron oxidation potential of the arene, nor with the expected stability of a sigma-complex derived from direct electrophilic aromatic substitution. It is argued that the observed rate constants correspond to initial formation of a pi-complex between the arylnitrenium ion and the arene trap. This complex then forms the observed products. PMID- 11929246 TI - Catalytic regioselective sulfonylation of alpha-chelatable alcohols: scope and mechanistic insight. AB - This paper describes a convenient protocol for the regioselective sulfonylation of alpha-chelatable alcohols. Typically, the reaction of alpha-heterosubstituted alcohols with 1 equiv of p-TsCl and 1 equiv of Et(3)N in the presence of 2 mol % of Bu(2)SnO leads to rapid, regioselective, and exclusive monotosylation. The pK(a) of the amine was correlated to the reaction rate. A plausible mechanism for this reaction has been proposed on the basis of (119)Sn NMR studies. PMID- 11929247 TI - Regulation of one-electron oxidation rate of guanine by base pairing with cytosine derivatives. AB - Effects of base pairing on the one-electron oxidation rate of guanine derivatives, guanine, 8-bromoguanine, and 8-oxo-7,8-dihydroguanine have been studied. The one-electron oxidation rate of guanine derivatives was determined by triplet-quenching experiments, using N,N'-dibutylnaphthaldiimide (NDI) in the triplet excited state (3NDI*) and fullerene (C(60)) in the triplet excited state ((3)C(60*)) as oxidants. In all three guanine derivatives studied here, acceleration of the one-electron oxidation was observed upon hydrogen bonding with cytosine, which demonstrates lowering of the oxidation potential of guanine derivatives by base pairing with cytosine. When a methyl or bromo group was introduced to the C5 position of cytosine, acceleration or suppression of the one electron oxidation relative to the guanine:cytosine base pair was observed, respectively. The results demonstrate that the one-electron oxidation rate of guanine in DNA can be regulated by introducing a substituent on base pairing cytosine. PMID- 11929248 TI - Solid-phase synthesis of oligosaccharides and on-resin quantitative monitoring using gated decoupling (13)C NMR. AB - A general strategy for solid-phase oligosaccharide synthesis capable of nondestructive quantitative monitoring has been developed. The synthesis was carried out on TentaGel using thioglycosides as glycosylating agents and dimethylthiomethylsulfonium triflate as the activator. An acylsulfonamide linker was introduced to cleave the oligosaccharide from the resin. The solid-phase reactions were monitored quantitatively by using the inverse gated decoupling technique of (13)C NMR, where two (13)C-enriched markers were used to monitor the reactions: one was (13)C-enriched glycine incorporated as a part of the linker and as an internal standard, and the other was a (13)C-enriched acetyl group used as a protecting group of the glycosylation reagent. A representative synthesis of sialyl Lewis X branched tetrasaccharide was demonstrated. PMID- 11929249 TI - Unusual temperature dependence in the cis/trans-oxetane formation discloses competitive syn versus anti attack for the Paterno-Buchi reaction of triplet excited ketones with cis- and trans-cylooctenes. Conformational control of diastereoselectivity in the cyclization and cleavage of preoxetane diradicals. AB - Toluene-d(8) solutions of cis- and trans-cyclooctene (cis- and trans-1a) as well as (Z)- and (E)-1-methylcyclooctene (cis- and trans-1b) have been irradiated at temperatures between -95 and +110 degrees C in the presence of benzophenone (BP) to afford mixtures of the cis- and trans-configured oxetanes 2a,b and the regioisomeric 2b'. Correspondingly, benzoquinone (BQ) gave with cis- and trans-1a the cycloadducts cis- and trans-3a. The cis/trans diastereomeric ratios of the [2 + 2]-cycloadducts 2 and 3 display a strong temperature dependence; with cis- and trans-1a or cis-1b as starting materials, the diastereoselectivity of the oxetane formation is high at low temperature, under preservation of the initial cyclooctene configuration. With increasing temperature, the cis diastereoselectivity decreases continuously for the cis-cyclooctenes; in the case of the cis-1a, the diastereoselectivity is even switched to trans (cis/trans ca. 20:80) at very high temperatures. For the strained trans-1a, the trans-oxetanes are strongly preferred over the entire temperature range, with only minor leakage (up to 10%) to the cis-oxetanes at very high temperatures. Oxetane formation is accompanied by nonthermal trans-to-cis isomerization of the cyclooctene. The methyl-substituted trans-1b constitutes an exceptional substrate; it displays cis diastereoselectivity in the [2 + 2] photocycloaddition at low temperatures for both regioisomers 2b and 2b', and the trans selectivity increases at moderate temperature (cis/trans = 4:96), to decrease again at high temperature, especially for the minor regioisomer 2b'. This complex temperature behavior of the cis/trans diastereoselectivity may be rationalized in terms of the triplet-diradical mechanism of the Paterno-Buchi reaction. We propose that the cyclooctene may be competitively attacked by the triplet-excited ketone from the higher (syn) or the less (anti) substituted side; such syn and anti trajectories have hitherto not been considered. To account for the unusual temperature behavior in the diastereoselectivity of the present [2 + 2] photocycloaddition, we suggest that temperature-dependent conformational changes of the resulting triplet preoxetane diradicals compete with their cyclization to the cis/trans-oxetane diastereomers and retro cleavage to the cis-cyclooctene. PMID- 11929250 TI - Stereoselective synthesis of tetrahydrofurans via formal [3+2]-cycloaddition of aldehydes and allylsilanes. Formal total synthesis of the muscarine alkaloids (-) allomuscarine and (+)-epimuscarine. AB - The stereoselective synthesis of tetrahydrofurans was achieved by formal [3+2] cycloaddition of allyl and crotylsilanes with alpha-triethylsilyloxy aldehydes. The scope of the reaction was examined by using different alpha-substituted aldehydes and different substituents on the silicon. Tamao oxidation of the products resulted in formation of diols that are easily functionalized allowing an entry to natural products synthesis. The formal total synthesis of the muscarine alkaloids (-)-allomuscarine and (+)-epimuscarine was achieved. PMID- 11929251 TI - Mechanism of propylene insertion using bis(phenoxyimine)-based titanium catalysts: an unusual secondary insertion of propylene in a group IV catalyst system. AB - A highly regioselective secondary enchainment of propylene in a group IV catalyst system is reported. End-group analysis of polypropylene formed using the phenoxyimine-based titanium catalysts revealed a reversal in the regioselectivity of insertion for this class of catalysts. To the best of our knowledge, bis(phenoxyimine)-based titanium complexes are the only known group IV catalysts that insert propylene with exclusive 2,1-regiochemistry. Insertion of propylene into the initiating titanium hydride occurs with high 1,2-regiochemistry. Subsequent insertions into primary titanium alkyls are regiorandom, while insertions into secondary titanium alkyls proceed with high 2,1-regioselectivity. Cyclopolymerization and ethylene/propylene copolymerization strategies are employed to support this proposal. PMID- 11929252 TI - Antihydrophobic cosolvent effects for alkylation reactions in water solution, particularly oxygen versus carbon alkylations of phenoxide ions. AB - Antihydrophobic cosolvents such as ethanol increase the solubility of hydrophobic molecules in water, and they also affect the rates of reactions involving hydrophobic surfaces. In simple reactions of hydrocarbons, such as the Diels Alder dimerization of 1,3-cyclopentadiene, the rate and solubility data directly reflect the geometry of the transition state, in which some hydrophobic surface becomes hidden. In reactions involving polar groups, such as alkylations of phenoxide ions or S(N)1 ionizations of alkyl halides, cosolvents in water can have other effects as well. However, solvation of hydrophobic surfaces is still important. By the use of structure-reactivity relationships, and comparing the effects of ethanol and DMSO as solvents, it has been possible to sort out these effects. The conclusions are reinforced by an ab initio computer model for hydrophobic solvation. The result is a sensible transition state for phenoxide ion as a nucleophile, using its oxygen n electrons to avoid loss of conjugation. The geometry of alkylation of aniline is very different, involving packing (stacking) of the aniline ring onto the phenyl ring of a benzyl group in the benzylation reaction. The alkylation of phenoxide ions by benzylic chlorides can occur both at the phenoxide oxygen and on ortho and para positions of the ring. Carbon alkylation occurs in water, but not in nonpolar organic solvents, and it is observed only when the phenoxide has at least one methyl substituent ortho, meta, or para. The effects of phenol substituents and of antihydrophobic cosolvents on the rates of the competing alkylation processes indicate that in water the carbon alkylation involves a transition state with hydrophobic packing of the benzyl group onto the phenol ring. The results also support our conclusion that oxygen alkylation uses the n electrons of the phenoxide oxygen as the nucleophile and does not have hydrophobic overlap in the transition state. The mechanisms and explanations for competing oxygen and carbon alkylations differ from previous proposals by others. PMID- 11929253 TI - The boron-catalyzed polymerization of dimethylsulfoxonium methylide. A living polymethylene synthesis. AB - Trialkyl and aryl organoboranes catalyze the polymerization of dimethylsulfoxonium methylide (1). The product of the polymerization is a tris polymethylene organoborane. Oxidation affords linear telechelic alpha-hydroxy polymethylene. The polymer molecular weight was found to be directly proportional to the stoichiometric ratio of ylide/borane, and polydispersities as low as 1.01 1.03 have been realized. Although oligomeric polymethylene has been the most frequent synthetic target of this method, polymeric star organoboranes with molecular weights of 1.5 million have been produced. The average turnover frequency at 120 degrees C in 1,2,4,5-tetrachlorobenzene/toluene is estimated at >6 x 10(6) g of polymethylene (mol boron)(-1) h(-1). The mechanism of the polyhomologation reaction involves initial formation of a zwitterionic organoborane.ylide complex which breaks down in a rate-limiting 1,2-alkyl group migration with concomitant expulsion of a molecule of DMSO. The reaction was found to be first order in the borane catalyst and zero order in ylide. DMSO does not interfere with the reaction. The temperature dependence of the reaction rate yielded the following activation energy parameters (toluene, DeltaH(++) = 23.2 kcal/mol, DeltaS(++) = 12.6 cal deg/mol, DeltaG(++) = 19.5 kcal/mol; THF, DeltaH(++) = 26.5 kcal/mol, DeltaS(++) = 21.5 cal deg/mol, DeltaG(++) = 20.1 kcal/mol). PMID- 11929254 TI - Enantio- and diastereoselective stepwise cyclization of polyprenoids induced by chiral and achiral LBAs. A new entry to (-)-ambrox, (+)-podocarpa-8,11,13-triene diterpenoids, and (-)-tetracyclic polyprenoid of sedimentary origin. AB - An enantio- and diastereoselective stepwise cyclization of polyprenoids induced by Lewis acid-assisted chiral Bronsted acids (chiral LBAs) and achiral LBAs is described. In particular, the absolute stereocontrol in the initial cyclization of polyprenoids to form an A-ring induced by chiral LBAs and the importance of the nucleophilicity of the internal terminator in polyprenoids for the relative stereocontrol in subsequent cyclization are demonstrated. (-)-Ambrox was synthesized via the enantioselective cyclization of (E,E)-homofarnesyl triethylsilyl ether with tin(IV) chloride-coordinated (R)-2-(o-fluorobenzyloxy) 2'-hydroxy-1,1'-binaphthyl ((R)-BINOL-o-FBn) and subsequent diastereoselective cyclization with CF(3)CO(2)H.SnCl(4) as key steps. Protection of (E,E) homofarnesol by a triethylsilyl group increased the enantioselectivity of chiral LBA-induced cyclization and both the chemical yield and diastereoselectivity in the subsequent cyclization. The enantioselective cyclization of homo(polyprenyl)arenes possessing an aryl group was also induced by (R)-BINOL-o FBn.SnCl(4). Several optically active podocarpa-8,11,13-triene diterpenoids and ( )-tetracyclic polyprenoid of sedimentary origin were synthesized (75-80% ee) by the enantioselective cyclization of homo(polyprenyl)benzene derivatives induced by (R)-BINOL-o-FBn.SnCl(4) and subsequent diastereoselective cyclization induced by BF(3).Et(2)O/EtNO(2) or CF(3)CO(2)H .SnCl(4). PMID- 11929255 TI - The first total synthesis of lipid II: the final monomeric intermediate in bacterial cell wall biosynthesis. AB - Bacterial peptidoglycan is composed of a network of beta-[1,4]-linked glyan strands that are cross-linked through pendant peptide chains. The final product, the murein sacculus, is a single, covalently closed macromolecule that precisely defines the size and shape of the bacterial cell. The recent increase in bacterial resistance to cell wall active agents has led to a resurgence of activity directed toward improving our understanding of the resistance mechanisms at the molecular level. The biosynthetic enzymes and their natural substrates can be invaluable tools in this endeavor. While modern experimental techniques have led to isolation and purification of the biosynthetic enzymes utilized in peptidoglycan biosynthesis, securing useful quantities of their requisite substrates from natural substrates has remained problematic. In an effort to address this issue, we report the first total synthesis of lipid II (4), the final monomeric intermediate utilized by Gram positive bacteria for peptidoglycan biosynthesis. PMID- 11929256 TI - Synthesis, properties, and reactivity of cocaine benzoylthio ester possessing the cocaine absolute configuration. AB - One aspect of immunopharmacotherapy for cocaine abuse involves the use of a catalytic monoclonal antibody (mAb) to degrade cocaine via hydrolysis of the benzoate ester. A cocaine benzoylthio ester analogue provides a means to implement high-throughput selection strategies to potentially isolate mAbs with high activity. The required analogue was synthesized starting from (-)-cocaine hydrochloride and possessed the cocaine absolute configuration. Key points in the preparation were the introduction of the sulfur atom at C-3 via a bromomagnesium thiolate addition to the exo face of anhydroecgonine, separation of C-2 diastereomers, recycling of a C-2 thio ester byproduct, and formation of the necessary C-2 methyl and C-3 benzoylthio esters. Effects resulting from the lower electronegativity and greater hydrophobicity of sulfur compared to oxygen were observed. These characteristics could result in interesting drug properties. Furthermore, the analogue was found to be a substrate for catalytic mAbs that hydrolyze cocaine as monitored by HPLC and also spectrophotometry by coupling cleavage of the benzoylthio ester to the disulfide exchange with Ellman's reagent. Screening antibody libraries with the new cocaine analogue using the spectroscopic assay provides an avenue for the high-throughput identification of catalysts that efficiently breakdown cocaine. PMID- 11929258 TI - The first complete identification of a diastereomeric catalyst-substrate (alkoxide) species in an enantioselective ketone hydrogenation. Mechanistic investigations. AB - The enantioselective hydrogenations of the dialkyl 3,3-dimethyloxaloacetate ketone substrates (2, 3, and 4; alkyl = Me, (i)Pr, and (t)Bu, respectively) were catalyzed by [Ru((R)-BINAP)(H)(MeCN)(n)(sol)(3-n)](BF(4)) (1, n = 0-3, sol = THF or MeOH, (R)-BINAP = (R)-2,2'-bis(diphenylphosphino)-1,1'-binaphthyl) in up to 82% ee (R). Reaction of the active catalyst 1 with 1 equiv of substrate (2, 3, or 4) in THF or MeOH solution formed the diastereomeric catalyst-alkoxide complexes [Ru((R)-BINAP)(MeCN)(OCH(CO(2)R)-(C(CH(3))(2)CO(2)R))](BF(4)) (5/6 R = Me, 8/9 R = (i)Pr, and 10 R = (t)Bu, respectively) via hydride addition to the ketone carbonyl carbon and ruthenium addition to oxygen. The absolute configurations at the alkoxide groups ((R)- for the major diastereomers 5, 8, and 10) were determined via cleavage of the ruthenium-alkoxide bond with 1 equiv of HBF(4).OEt(2). The solution structures of the major diastereomer catalyst alkoxide complexes (5, 8, and 10) were unambiguously determined by variable temperature NMR spectroscopy. The major diastereomers (5, 8, and 10) had the same absolute configuration as the major product enantiomers from the catalytic hydrogenation of 2, 3, and 4 with 1 as catalyst. The ratio of major to minor alkoxide diastereomers was similar to the ee of the catalytic hydrogenation. The catalyst-alkoxide complexes are formed at temperatures as low as -30 degrees C with no other precursors or intermediates observed by NMR showing that ketone hydride insertion is likely not the turnover limiting step of the catalytic hydrogenation. Results from the stoichiometric hydrogenolysis of 5/6, 8/9, or 10 indicate that their formation is rapid and only partially reversible prior to the irreversible hydrogenolysis of the ruthenium-oxygen bond. The stereoselectivities of the formation and hydrogenolysis of 5/6, 8/9, and 10 sum up to equal the stereoselectivities of the respective catalytic hydrogenations of 2, 3, and 4. The rates of the hydrogenolysis were consistent with these diastereomers being true catalytic intermediates. PMID- 11929257 TI - A general nickel-catalyzed hydroamination of 1,3-dienes by alkylamines: catalyst selection, scope, and mechanism. AB - A simple colorimetric assay of various transition-metal catalysts showed that the combination of DPPF, Ni(COD)(2), and acid is a highly active catalyst system for the hydroamination of dienes by alkylamines to form allylic amines. The scope of the reaction is broad; various primary and secondary alkylamines react with 1,3 dienes in the presence of these catalysts. Detailed mechanistic studies revealed the individual steps involved in the catalytic process. These studies uncovered unexpected thermodynamics for the addition of amines to pi-allyl nickel complexes: instead of the thermodynamics favoring the reaction of a nickel allyl with an amine to form an allylic amine, the thermodynamics favored reaction of a nickel(0) complex with allylic amine in the presence of acid to form a Ni(II) allyl. The realization of these thermodynamics led us to the discovery that nickel and some palladium complexes in the presence or absence of acid catalyze the exchange of the amino groups of allylic amines with free amines. This exchange process was used to reveal the relative thermodynamic stabilities of various allylic amines. In addition, this exchange reaction leads to racemization of allylic amines. Therefore, the relative rate for C-N bond formation and cleavage influences the enantioselectivity of diene hydroaminations. PMID- 11929259 TI - Acetylide-bridged organometallic oligomers via the photochemical metathesis of methyl-iron(II) complexes. AB - The acetylido methyl iron(II) complexes, cis/trans-[Fe(dmpe)(2)(C[triple bond]CR)(CH(3))] (1) and trans-[Fe(depe)(2)(C[triple bond]CR)(CH(3))] (2) (dmpe = 1,2-dimethylphoshinoethane; depe = 1,2-diethylphosphinoethane), were synthesized by transmetalation from the corresponding alkyl halide complexes. Acetylido methyl iron(II) complexes were also formed by transmetalation from the chloride complexes, trans-[Fe(dmpe)(2)(C[triple bond]CR)(Cl)] or trans [Fe(depe)(2)(C[triple bond]CR)(Cl)]. The structure of trans-[Fe(dmpe)(2)(C[triple bond]CC(6)H(5))(CH(3))] (1a) was determined by single-crystal X-ray diffraction. The methyl acetylido iron complexes, [Fe(dmpe)(2)(C[triple bond]CR)(CH(3))] (1), are thermally stable in the presence of acetylenes; however, under UV irradiation, methane is lost with the formation of a metal bisacetylide. Photochemical metathesis of cis- or trans-[Fe(dmpe)(2)(CH(3))(C[triple bond]CR)] (R = C(6)H(5) (1a), 4-C(6)H(4)OCH(3) (1b)) with terminal acetylenes was used to selectively synthesize unsymmetrically substituted iron(II) bisacetylide complexes of the type trans-[Fe(dmpe)(2)(C[triple bond]CR)(C[triple bond]CR')] [R = Ph, R' = Ph (6a), 4-CH(3)OC(6)H(4) (6b), (t)()Bu (6c), Si(CH(3))(3) (6d), (CH(2))(4)C[triple bond]CH (6e); R = 4-CH(3)OC(6)H(4), R' = 4-CH(3)OC(6)H(4), (6g), (t)()Bu (6h), (CH(2))(4)C[triple bond]CH (6i), adamantyl (6j)]. The structure of the unsymmetrical iron(II) bisacetylide complex trans [Fe(dmpe)(2)(C[triple bond]CC(6)H(5))(C[triple bond]CC(6)H(4)OCH(3))] (6b) was determined by single-crystal X-ray diffraction. The photochemical metathesis of the bis-acetylene, 1,7-octadiyne, with trans-[Fe(dmpe)(2)(CH(3))(C[triple bond]CPh)] (1a), was utilized to synthesize the bridged binuclear species trans,trans-[(C(6)H(5)C[triple bond]C)Fe(dmpe)(2)(mu-C[triple bond]C(CH(2))(4)C[triple bond]C)Fe(dmpe)(2)(C[triple bond]CC(6)H(5))] (11). The trinuclear species trans,trans,trans-[(C(6)H(5)C[triple bond]C)Fe(dmpe)(2)(mu C[triple bond]C(CH(2))(4)C[triple bond]C)Fe(dmpe)(2)(mu-C[triple bond]C(CH(2))(4)C[triple bond]C)Fe(dmpe)(2)(C[triple bond]CC(6)H(5))] (12) was synthesized by the photochemical reaction of Fe(dmpe)(2)(C[triple bond]CPh)(C[triple bond]C(CH(2))(4)C[triple bond]CH) (6e) with Fe(dmpe)(2)(CH(3))(2). Extended irradiation of the bisacetylide complexes with phenylacetylene resulted in insertion of the terminal alkyne into one of the metal acetylide bonds to give acetylide butenyne complexes. The structure of the acetylide butenyne complex, trans-[Fe(dmpe)(2)(C[triple bond]CC(6)H(4)OCH(3))(eta(1)-C(C(6)H(5))=CH(C[triple bond]CC(6)H(4)OCH(3)))] (9a) was determined by single-crystal X-ray diffraction. PMID- 11929260 TI - Synthesis, structure, and bonding properties of 5-carbaphosphatranes: a new class of main group atrane. AB - 1-Hydro-5-carbaphosphatrane (1) and 1-methyl-5-carbaphosphatrane (2), the first 5 carbon analogues of phosphatranes, were synthesized by a demethylation reaction of cyclic phosphinate 3. X-ray analysis revealed that 1 has a typical trigonal bipyramidal structure with hydrogen and carbon atoms at the apical position and three oxygen atoms at the equatorial positions, indicating that 1 is a phosphorane in the perfectly "anti-apicophilic" arrangement. Apical P-C and P-H bond lengths were 1.921(2) and 1.38(2) A, respectively. The (1)J(PH) value of 1 and the (1)J(PC)(P-CH(3)) value of 2 were 852 and 215 Hz, respectively, which are extraordinarily large for the apical coupling constants of phosphoranes, but close to those of the reported phosphatranes with a 5-nitrogen atom. IR and Raman spectra are also reported. Force constant calculations indicate the transannular bond in carbaphosphatrane is 3 times stronger than in silatrane, due to its covalent character. PMID- 11929261 TI - Chemical control of phase transformation kinetics in periodic silica/surfactant composites. AB - Control of phase stability is investigated through control of silica chemistry in ordered silica/surfactant composites under hydrothermal conditions. The composites were hydrothermally treated in pH 9 through pH 11 buffers while using in situ real time X-ray diffraction to follow a p6mm hexagonal-to-lamellar structural transition. The data were analyzed using both isothermal and nonisothermal (temperature-ramped) kinetics to determine activation energies. It was found that the most mildly basic conditions utilized (pH 9), which favor silica condensation, best inhibit the phase transition and thus produce the most kinetically stable composites. High-pH treatment, conversely, allows for the most facile rearrangements. Condensation occurring during composite synthesis rather than during hydrothermal treatment has a much smaller effect on phase stability, probably because much of the condensation that occurs during synthesis is random and not optimally coupled to the nanoscale architecture. Materials that start out poorly condensed, by contrast, can be extensively hydrothermally modified so that the final material has an inorganic framework with a highly uniform silica density; this provides the maximum resistance to transformation and the highest kinetic stability. In all cases, very good agreement is found between the results of isothermal and nonisothermal kinetic methods. The trends across pHs indicate that both isothermal and nonisothermal measurements are accurate and that differences between them are meaningful and represent physical differences in the transforming materials resulting from the different heating processes. PMID- 11929262 TI - Single-molecule magnets: a new family of Mn(12) clusters of formula [Mn(12)O(8)X(4)(O(2)CPh)(8)L(6)]. AB - The reaction of (NBu(n)(4))[Mn(8)O(6)Cl(6)(O(2)CPh)(7)(H(2)O)(2)] (1) with 2 (hydroxymethyl)pyridine (hmpH) or 2-(hydroxyethyl)pyridine (hepH) gives the Mn(II)(2)Mn(III)(10) title compounds [Mn(12)O(8)Cl(4)(O(2)CPh)(8)(hmp)(6)] (2) and [Mn(12)O(8)Cl(4)(O(2)CPh)(8)(hep)(6)] (3), respectively, with X = Cl. Subsequent reaction of 3 with HBr affords the Br(-) analogue [Mn(12)O(8)Br(4)(O(2)CPh)(8)(hep)(6)] (4). Complexes 2.2Et(2)O.4CH(2)Cl(2), 3.7CH(2)Cl(2), and 4.2Et(2)O.1.4CH(2)Cl(2) crystallize in the triclinic space group P1, monoclinic space group C2/c, and tetragonal space group I4(1)/a, respectively. Complexes 2 and 3 represent a new structural type, possessing isomeric [Mn(III)(10)Mn(II)(2)O(16)Cl(2)] cores but with differing peripheral ligation. Complex 4 is essentially isostructural with 3. A magnetochemical investigation of complex 2 reveals an S = 6 or 7 ground state and frequency dependent out-of-phase signals in ac susceptibility studies that establish it as a new class of single-molecule magnet. These signals occur at temperatures higher than those observed for all previously reported single-molecule magnets that are not derived from [Mn(12)O(12)(O(2)CR)(16)(H(2)O)(x)]. A detailed investigation of forms of complex 2 with different solvation levels reveals that the magnetic properties of 2 are extremely sensitive to the latter, emphasizing the importance to the single-molecule magnet properties of interstitial solvent molecules in the samples. In contrast, complexes 3 and 4 are low-spin molecules with an S = 0 ground state. PMID- 11929263 TI - Pi-complexation of biphenyl, naphthalene, and triphenylene to trimeric perfluoro ortho-phenylene mercury. Formation of extended binary stacks with unusual luminescent properties. AB - Trimeric perfluoro-ortho-phenylene mercury (1) crystallizes from CS(2) as a pure compound. In the crystal, 1 forms staggered cofacial dimers (centroid distance of 3.38 A). In the dimer, the individual components are associated via long mercury pi interactions (3.443 < Hg...C < 3.650 A). Interestingly, this arrangement leads to the existence of relatively short intermolecular mercury-mercury distances (3.811 < Hg...Hg < 4.093 A). In this form, compound 1 is photoluminescent and exhibits a broad emission band with a maximum at 440 nm and a shoulder at 530 nm. Compound 1 interacts with biphenyl, naphthalene, or triphenylene to form 1.biphenyl (2), 1.naphthalene (3), and 1.triphenylene (4), respectively. These adducts have been characterized by elemental analysis and X-ray crystallography. Their structure reveals the existence of stacks in which molecules of 1 and molecules of arenes alternate. In each stack, secondary pi-interactions occur between the arene and the mercury centers of 1. The resulting Hg...C distances range from 3.25 to 3.55 A and are within the sum of the van der Waals radii. They reflect the presence of secondary polyhapto-pi interactions occurring between the electron-rich aromatic molecules and the acidic mercury centers. In the case of the triphenylene adduct 4, a arene-fluoroarene interaction is also observed (centroid distance of 3.605 A). Compounds 2-4 are photoluminescent. The emission observed for 2 and 3 corresponds to the phosphorescence of the aromatic substrate and suggests the occurrence of a mercury heavy atom effect. In the case of 4, the emission appears at longer wavelengths than those typically observed for triphenylene. PMID- 11929264 TI - Arene-mercury complexes stabilized by aluminum and gallium chloride: catalysts for H/D exchange of aromatic compounds. AB - Dissolution of Hg(arene)(2)(MCl(4))(2) [arene = C(6)H(5)Me, C(6)H(5)Et, o C(6)H(4)Me(2), C(6)H(3)-1,2,3-Me(3); M = Al, Ga] in C(6)D(6) results in a rapid H/D exchange and the formation of the appropriate d(n)-arene and C(6)D(5)H. H/D exchange is also observed between C(6)D(6) and the liquid clathrate ionic complexes, [Hg(arene)(2)(MCl(4))][MCl(4)], formed by dissolution of HgCl(2) and MCl(3) in C(6)H(6), m-C(6)H(4)Me(2), or p-C(6)H(4)Me(2). The H/D exchange reaction is found to be catalytic with respect to Hg(arene)(2)(MCl(4))(2) and independent of the initial arene ligand. Reaction of a 1:1 ratio of C(6)H(5)Me and C(6)D(6) with <0.1 mol % Hg(C(6)H(5)Me)(2)(MCl(4))(2) results in an equilibrium mixture of all isotopic isomers: C(6)H(5-x)D(x)Me and C(6)D(6-x)H(x) (x = 0-5). DFT calculations on the model system, Hg(C(6)H(6))(2)(AlCl(4))(2) and [Hg(C(6)H(6))(2)(AlCl(4))](+), show that the charge on the carbon and proton associated with the shortest Hg...C interactions is significantly higher than that on uncomplexed benzene or HgCl(2)(C(6)H(6))(2). The protonation of benzene by either Hg(C(6)H(6))(2)(AlCl(4))(2) or [Hg(C(6)H(6))(2)(AlCl(4))](+) was calculated to be thermodynamically favored in comparison to protonation of benzene by HO(2)CCF(3), a known catalyst for arene H/D exchange. Arene exchange and intramolecular hydrogen transfer reactions are also investigated by DFT calculations. PMID- 11929265 TI - Ru(II) and Os(II) nucleosides and oligonucleotides: synthesis and properties. AB - A general and versatile method for the site-specific incorporation of polypyridine Ru(II) and Os(II) complexes into DNA oligonucleotides using solid phase phosphoramidite chemistry is reported. Novel nucleosides containing a [(bpy)(2)M(3-ethynyl-1,10-phenanthroline)](2+) (M = Ru, Os) metal center covalently attached to the 5-position in 2'-deoxyuridine are synthesized, and their electrochemical as well as photophysical properties are studied. The Ru(II) nucleoside exhibits a rather long-lived excited state in phosphate buffer pH 7.0 (tau = 1.08 micros) associated with a relatively high emission quantum efficiency (phi = 0.051). The solvent dependence of the absorption and emission spectra is consistent with an emissive MLCT state where charge localization takes place on the extended heterocycle-linked phenanthroline. In contrast, the Os(II) containing nucleoside is quite nonemissive in aqueous environment (tau = 0.027 micros, phi = 1 x 10(-4)). The metal-containing nucleosides are converted into their phosphoramidites and are utilized for the high-yield preparation of modified oligonucleotides. The novel oligonucleotides, characterized by absorption and emission spectroscopy, enzymatic digestion, and electrophoresis, form stable duplexes. Circular dichroism spectra confirm that the global conformation of the double helix is not altered by the presence of these polypyridyl complexes in the major groove. Metal-containing phosphoramidites with predetermined absolute configuration at the octahedral coordination center are synthesized and utilized for the synthesis of diasteromerically pure metal containing DNA oligonucleotides. Emission spectroscopy suggests a higher protection of the Delta metal center from the bulk solvent and better accommodation within the major groove. PMID- 11929266 TI - Enzymatic GTP hydrolysis: insights from an ab initio molecular dynamics study. AB - Ab initio methods were used to shed light on fundamental aspects of the enzymatic mechanism of guanosine triphosphate hydrolysis in the Cdc42/Cdc42GAP complex. The calculations focused on the nucleophilic addition of the catalytic water molecule to the gamma-phosphate phosphorus atom. A large model system was required to correctly reproduce the electrostatic properties on the active site. The model turned out to reproduce most of the electrostatic field of the biological complex at the reactants. Our calculations established the H-bond pattern of the catalytic water (WAT), which turned out to interact with Q61 and T35, in the most stable conformation. This ruled out the possibility that the catalytic water transferred its proton directly to the gamma-phosphate. Furthermore, the calculations suggested that the electronic structure of WAT was very different from that in the bulk. Finally, this study showed that during the reaction, WAT transferred a proton to Gln61, consistent with the available X-ray data on a transition-state analogue/enzyme complex(19) and with the decrease of activity in the Q61E mutant. PMID- 11929267 TI - Second-sphere contributions to substrate-analogue binding in iron(III) superoxide dismutase. AB - A combination of spectroscopic and computational methods has been employed to explore the nature of the yellow and pink low-temperature azide adducts of iron(III) superoxide dismutase (N(3)-FeSOD), which have been known for more than two decades. Variable-temperature variable-field magnetic circular dichroism (MCD) data suggest that both species possess similar ferric centers with a single azide ligand bound, contradicting previous proposals invoking two azide ligands in the pink form. Complementary data obtained on the azide complex of the Q69E FeSOD mutant reveal that relatively minor perturbations in the metal-center environment are sufficient to produce significant spectral changes; the Q69E N(3) FeSOD species is red in color at all temperatures. Resonance Raman (RR) spectra of the wild-type and Q69E mutant N(3)-FeSOD complexes are consistent with similar Fe-N(3) units in all three species; however, variations in energies and relative intensities of the RR features associated with this unit reveal subtle differences in (N(3)(-))-Fe(3+) bonding. To understand these differences on a quantitative level, density functional theory and semiempirical INDO/S-CI calculations have been performed on N(3)-FeSOD models. These computations support our model that a single azide ligand is present in all three N(3)-FeSOD adducts and suggest that their different appearances reflect differences in the Fe-N-N bond angle. A 10 degrees increase in the Fe-N-N bond angle is sufficient to account for the spectral differences between the yellow and pink wild-type N(3) FeSOD species. We show that this bond angle is strongly affected by the second coordination sphere, which therefore might also play an important role in orienting incoming substrate for reaction with the FeSOD active site. PMID- 11929268 TI - Energetics of radical transfer in DNA photolyase. AB - Charge separation and radical transfer in DNA photolyase from Escherichia coli is investigated by computing electrostatic free energies from a solution of the Poisson-Boltzmann equation. For the initial charge separation 450 meV are available. According to recent experiments [Aubert et al. Nature 2000, 405, 586 590] the flavin receives an electron from the proximal tryptophan W382, which consequently forms a cationic radical WH(*)(+)382. The radical state is subsequently transferred along the triad W382-W359-W306 of conserved tryptophans. The radical transfer to the intermediate tryptophan W359 is nearly isoenergetic (58 meV uphill); the radical transfer from the intermediate W359 to the distal W306 is 200 meV downhill in energy, funneling and stabilizing the radical state at W306. The resulting cationic radical WH(*)(+)306 is further stabilized by deprotonation, yielding the neutral radical W(*)306, which is 214 meV below WH(*)(+)306. The time scale of the charge recombination process yielding back the resting enzyme with FADH(*) is governed by reprotonation of W306, with a calculated lifetime of 1.2 ms that correlates well with the measured lifetime of 17 ms. In photolyase from Anacystis nidulans the radical state is partially transferred to a tyrosine [Aubert et al. Proc. Natl. Acad. Sci. U.S.A. 1999, 96, 5423-5427]. In photolyase from Escherichia coli, there is a tyrosine (Y464) close to the distal tryptophan W306 that could play this role. We show that this tyrosine cannot be involved in radical transfer, because the electron transfer from tyrosine to W306 is much too endergonic (750 meV) and a direct hydrogen transfer is likely too slow. Coupling of specific charge states of the tryptophan triad with protonation patterns of titratable residues of photolyase is small. PMID- 11929269 TI - A new interpretation of the IR bands of supported Rh(I) monocarbonyl complexes. AB - The characteristic CO vibrational frequency of supported monocarbonyl complexes Rh(I)CO, at 2014 and 1984 cm(-1) on dealuminated Y zeolite and alumina, respectively, is lower than the frequencies of both the symmetric and the antisymmetric CO normal modes of the corresponding stable supported Rh(I) dicarbonyls. The CO mode with a measured frequency between those of the symmetric and antisymmetric CO frequencies of the dicarbonyls, previously assigned to rhodium monocarbonyl, is reassigned to mixed carbonyl dihydrogen complexes Rh(H(2))(CO) or Rh(H)(2)(CO). This reassignment is based on a critical analysis of reported experimental data, supplemented by quantum chemical calculations. PMID- 11929270 TI - Switching the inside and the outside of aggregates of water-soluble block copolymers with double thermoresponsivity. AB - Water-soluble block copolymers were prepared from the nonionic monomer N isopropylacrylamide (NIPA) and the zwitterionic monomer 3-[N-(3 methacrylamidopropyl)-N,N-dimethyl]ammoniopropane sulfonate (SPP) by sequential free radical polymerization via the RAFT process. Such block copolymers with two hydrophilic blocks exhibit double thermoresponsive behavior in water: the poly NIPA block shows a lower critical solution temperature, whereas the poly-SPP block exhibits an upper critical solution temperature. Appropriate design of the block lengths leads to block copolymers which stay in solution in the full temperature range between 0 and 100 degrees C. Both blocks of these polymers dissolve in water at intermediate temperatures, whereas at high temperatures, the poly-NIPA block forms colloidal hydrophobic associates that are kept in solution by the poly-SPP block, and at low temperatures, the poly-SPP block forms colloidal polar aggregates that are kept in solution by the poly-NIPA block. In this way, colloidal aggregates which switch reversibly can be prepared in water, and without any additive, their "inside" to the "outside", and vice versa. The aggregates provide microdomains and surfaces of different character, which can be controlled by a simple thermal stimulus. PMID- 11929271 TI - Formation and decomposition of distonic o-, m-, and p-benzyne radical cations from photolysis of Mg(+)(o-, m-, p-C(6)H(4)F(2)). AB - Distonic o-, m-, and p-benzyne radical cations (1-3) have been generated by a novel photolysis reaction of mass-selected Mg(+)-difluorobenzene complexes. The energy required for the formation of these radical cations is within 2.2 eV. The formation of o-benzyne cation is most facile. The benzyne radical cations dissociate further to yield ethyne and 1,3-butadiyne radical cation as major products given a sufficient amount of energy. The whole process involves only a single photon, and is very efficient. The calculated threshold for the formation of 1,3-butadiyne radical cation from Mg(+)(o-C(6)H(4)F(2)) is about 4.6 eV, quite comparable with the experimental estimate. PMID- 11929272 TI - Determination of the diglyceride content in greek virgin olive oils and some commercial olive oils by employing (31)P NMR spectroscopy. AB - In this study, the diglyceride contents of 96 samples of virgin olive oils from the regions of Crete, Lesvos, Messinia, Pilion, Zakynthos, Halkidiki, and Ilia, 15 samples of commercial extra virgin and pure olive oils, and 3 samples each of refined olive oils and pomace oils were determined by a facile method introduced in a previous publication. This method is based on the phosphitylation of the free hydroxyls of the diglycerides with 2-chloro-4,4,5,5 tetramethyldioxaphospholane and the integration of the appropriate peaks in the (31)P NMR spectra. This preliminary study showed interesting trends in the diglyceride content of the virgin olive oils from the various regions of Greece that can be used as simple criteria to assess the olive oil characteristics. Analysis of variance has been carried out for the diglyceride content of each region in an attempt to detect possible differences in the diglyceride levels among the various regions. Finally, the relationship between the ratio of 1,2 diglycerides to the total amount of diglycerides and the total amount of diglycerides has been used to monitor the quality of virgin olive oils, commercial olive oils, refined olive oils, and pomace oils. PMID- 11929273 TI - Liquid chromatography-mass spectrometry of cis- and all-trans-lycopene in human serum and prostate tissue after dietary supplementation with tomato sauce. AB - Several epidemiological studies suggest a lower incidence of prostate cancer in men who routinely consume tomato products. Tomatoes are the primary dietary source of lycopene, which is among the most potent antioxidants of the carotenoids. Men with clinical stage T1 or T2 prostate adenocarcinoma were recruited (n = 32) and consumed tomato sauce based pasta dishes for 3 weeks (equivalent to 30 mg of lycopene per day) before radical prostectomy. Prostate tissue from needle biopsy just before intervention and prostectomy after supplementation from a subset of 11 subjects was evaluated for both total lycopene and lycopene geometrical isomer ratios. A gradient HPLC system using a C(18) column with UV-vis absorbance detection was used to measure total lycopene. Because the absorbance detector was insufficiently sensitive, HPLC with a C(30) column and positive ion atmospheric pressure chemical ionization mass spectrometric (LC-MS) detection was developed as a new assay to measure the ratio of lycopene cis/trans isomers in these samples. The limit of detection of the LC MS method was determined to be 0.93 pmol of lycopene on-column, and a linear response was obtained over 3 orders of magnitude. Total lycopene in serum increased 2.0-fold from 35.6 to 69.9 microg/dL (from 0.664 to 1.30 microM) as a result of dietary supplementation with tomato sauce, whereas total lycopene in prostate tissue increased 3.0-fold from 0.196 to 0.582 ng/mg of tissue (from 0.365 to 1.09 pmol/mg). all-trans-Lycopene and at least 14 cis-isomer peaks were detected in prostate tissue and serum. The mean proportion of all-trans-lycopene in prostate tissue was approximately 12.4% of total lycopene before supplementation but increased to 22.7% after dietary intervention with tomato sauce. In serum there was only a 2.8% but statistically significant increase in the proportion of all-trans-lycopene after intervention. These results indicate that short-term supplementation with tomato sauce containing primarily all-trans lycopene (83% of total lycopene) results in substantial increases in total lycopene in serum and prostate and a substantial increase in all-trans-lycopene in prostate but relatively less in serum. PMID- 11929274 TI - Microwave-assisted extraction (MAE)-acid hydrolysis of dithiocarbamates for trace analysis in tobacco and peaches. AB - A simple and rapid method is presented for the analysis of residues of ethylenebis(dithiocarbamate) (maneb, zineb, and mancozeb) and N,N dimethyldithiocarbamate (thiram and ziram) fungicides in dry tobacco leaves and peaches. Residues are extracted and hydrolyzed to CS(2) in a single step by use of microwave energy in a closed-vessel system while the evolved CS(2) trapped in a layer of iso-octane overlaying the reaction mixture is taken for gas chromatographic-flame photometric analysis. This combined extraction-hydrolysis step is carried out in 10 and 15 min for sets of 12 samples of tobacco and peach matrices, respectively. Total sample preparation time for GC analysis is 40 min. The limits of detection (LOD) are 0.005 mg/kg for thiram and ziram on peaches and 0.1 mg/kg for maneb, zineb, and mancozeb on tobacco. The respective LOD and limit of quantification (LOQ) levels in CS(2) equivalents are 0.003 and 0.006 mg/kg on peaches and 0.04 and 0.2 mg/kg on tobacco, respectively. Recoveries in the 0.01 60 mg/kg fortification range are 80-100% with respective relative standard deviations <20%. The method was used for the analysis of >3000 commercial tobacco samples including also different marketed cigarette brands. PMID- 11929275 TI - Application of solid/liquid extraction for the gravimetric determination of lipids in royal jelly. AB - Gravimetric lipid determination is a major parameter for the characterization and the authentication of royal jelly quality. A solid/liquid extraction was compared to the reference method, which is based on liquid/liquid extraction. The amount of royal jelly and the time of the extraction were optimized in comparison to the reference method. Boiling/rinsing ratio and spread of royal jelly onto the extraction thimble were identified as critical parameters, resulting in good accuracy and precision for the alternative method. Comparison of reproducibility and repeatability of both methods associated with gas chromatographic analysis of the composition of the extracted lipids showed no differences between the two methods. As the intra-laboratory validation tests were comparable to the reference method, while offering rapidity and a decrease in amount of solvent used, it was concluded that the proposed method should be used with no modification of quality criteria and norms established for royal jelly characterization. PMID- 11929276 TI - Antioxidant and cyclooxygenase activities of fatty acids found in food. AB - Several commercially available C-8 to C-24 saturated and unsaturated fatty acids (1-29) were assayed for cyclooxygenase-I (COX-I) and cyclooxygenase-II (COX-II) inhibitory and antioxidant activities. Among the saturated fatty acids tested at 60 microg mL(-1), there was an increase in antioxidant activity with increasing chain length from octanoic acid to myristic acid (C-8-C-14) and a decrease thereafter. All unsaturated fatty acids tested at 60 microg mL(-1) showed good antioxidant activity except for undecylenic acid (12), cis-5-dodecenoic acid (13), and nervonic acid (29). The highest inhibitory activities among the saturated fatty acids tested on cyclooxygenase enzymes COX-I and COX-II were observed for decanoic acid to lauric acid (3-5) at 100 microg mL(-1). Similarly, among the unsaturated fatty acids tested, the highest activities were observed for cis-8,11,14-eicosatrienoic acid (25) and cis-13,16-docosadienoic acid (27) at 100 microg mL(-1). PMID- 11929277 TI - Preparation of tetrahydroagathic acid: a serum metabolite of isocupressic acid, a cattle abortifacient in ponderosa pine. AB - Isocupressic acid (1) was used to synthetically prepare a mixture of (8S,13R,S) labda-15,19-dioic acid (tetrahydroagathic acid) (5) via a two-step oxidation procedure followed by hydrogenation of the double bonds at C13 and C8. Reduction of the C8,17 double bond was stereospecific producing only the 8S isomer and confirmed by the nOe interaction between the resulting C17 and C20 methyl groups. The 13R and 13S isomers of 5 were separated and analyzed by HPLC/MS, and (13S) tetrahydroagathic acid was isolated and identified by comparison to a standard prepared by hydrogenation of naturally occurring (13S)-dihydroagathic acid (4). (13R,S)-dihydroagathic acid was prepared by selective sodium metal-catalyzed hydrogenation of the C13,14 allylic double bond of agathic acid (3). The prepared compounds were then used as standards to confirm the presence of 4 and 5 and their respective 13R and 13S isomers in bovine serum samples. Tetrahydroagathic acid was shown to be the only metabolite detected in serum samples taken from a suspected cattle abortion case submitted for diagnosis; and, thus, 5 could be a valuable diagnostic marker for pine needle-induced abortions. PMID- 11929278 TI - Purification and characterization of two antifungal chitinases extracellularly produced by Bacillus amyloliquefaciens V656 in a shrimp and crab shell powder medium. AB - A Gram-positive bacterium with antagonistic activity was isolated from the soil. It has been identified as Bacillus amyloliquefaciens strain V656 on the basis of 16S ribosomal DNA analysis and standard bacteriological tests. B. amyloliquefaciens V656 produced antifungal enzymes when it was grown in a medium containing shrimp and crab shell powder (SCSP) of marine waste. The antifungal enzymes displayed chitinase activities. Two extracellular antifungal chitinases (FI and FII) were purified and characterized, and their molecular weights, isoelectric points, pH and thermal stabilities, and antifungal activities were determined. The characteristics of V656 chitinases are similar to those of the known bacterial chitinases in terms of their isoelectric points, thermal instabilities, and lack of lysozyme activity. In contrast to other known bacterial chitinases, the unique characteristics of V656 chitinases include extremely low molecular weights and nearly neutral optimum pH. Furthermore, this is the first report of the isolation of chitinases from B. amyloliquefaciens that are active against fungi. PMID- 11929279 TI - Purification and characterization of an antimicrobial chitinase extracellularly produced by Monascus purpureus CCRC31499 in a shrimp and crab shell powder medium. AB - Monascus purpureus CCRC31499 produced an antimicrobial chitinase when it was grown in a medium containing shrimp and crab shell powder (SCSP) of marine wastes. An extracellular antimicrobial chitinase was purified from the culture supernatant to homology. The chitinase had a molecular weight of approximately 81,000 and a pI of 5.4. The optimal pH, optimum temperature, and pH stability of the chitinase were pH 7, 40 degrees C, and pH 6-8, respectively. The activity of the chitinase was activated by Fe(2+) and strongly inhibited by Hg(2+). The unique characteristics of the purified chitinase include high molecular weight, nearly neutral optimum pH, protease activity, and antimicrobial activity with bacteria and fungal phytopathogens. This is also the first report of isolation of a chitinase from a Monascus species. PMID- 11929280 TI - Resistance of soybean vegetative storage proteins (S-VSPs) to proteolysis by rumen microorganisms. AB - Soybean vegetative storage proteins (S-VSPs) are lysine-rich and, hence, are potentially of high nutritive value for high productive ruminants. Using S-VSPs from wild-type soybean and from transgenic tobacco plants expressing either one of the two S-VSPs subunits (S-VSP alpha or S-VSP beta) or both, we tested their stability in cow rumen fluid under in situ conditions, using SDS-polyacrylamide gel electrophoresis. Proteolysis and degradation pattern of S-VSPs from transgenic tobacco leaves occurred relatively fast compared with that of wild type (WT) soybean plants. Comparing the two S-VSPs subunits expressed in transgenic plants, we found that S-VSP alpha was degraded much faster than S-VSP beta. The degradation pattern of S-VSPs in transgenic tobacco plants expressing both subunits resembled that of WT soybean. In contrast, the degradation pattern of transgenic tobacco plants expressing a single subunit was different. These finding suggest that the quaternary structure of S-VSPs may be an important factor determining their resistance to rumen degradation. Our results also suggest that the stability to rumen proteolysis of a given protein, when expressed in a transgenic plant, may not always be predictable and has to be verified. PMID- 11929281 TI - Changes in proanthocyanidin chain length in winelike model solutions. AB - Reactions of seed and skin proanthocyanidins in the presence or absence of (-) epicatechin were followed in winelike solutions over 53 days at 30 degrees C. Proanthocyanidins were separated from flavanol monomers by sequential elution from a Sep Pak cartridge, and changes in proanthocyanidin composition were monitored by thiolysis analysis of the proanthocyanidin fraction. In solutions containing no free (-)-epicatechin, trace amounts of monomers were released and important losses of proanthocyanidins were measured, but their average composition and mean degree of polymerization (mDP) were hardly modified. In the presence of (-)-epicatechin, the mDP value decreased and oligomeric proanthocyanidins accumulated throughout the incubation while losses of total units were dramatically reduced. Our data indicate that interflavanic bond cleavage of proanthocyanidins occurred under mild acidic conditions such as encountered in wine and that the resulting carbocation proceeded to unknown species. The latter reaction did not take place in the presence of (-) epicatechin. Epicatechin added to the intermediate carbocation, thus being incorporated as the end unit of a shorter proanthocyanidin chain. The results of this study are discussed in relation to the loss of astringency reported during wine aging. PMID- 11929282 TI - Fermentation of cottonseed and other feedstuffs in cattle rumen fluid. AB - Bovine rumen fluid was fermented anaerobically over 48 h with cottonseed, corn, alfalfa, or a mixture of these substrates in anaerobic mineral buffer. Samples taken at different incubation times were derivatized with n-butanol and subjected to gas chromatography and mass spectroscopy. No unusual fermentation end-products from the cottonseed substrate were detected. Cottonseed supported rumen fermentation at levels comparable to those of the other substrates. Major components were usually found in the decreasing order of acetate, propionate, butyrate, and valerate, although acetate and propionate concentrations decreased late in the alfalfa and mixed-feed fermentations, eventually allowing butyrate concentrations to exceed those of propionate. As expected, lactate was produced in high concentrations when corn was fermented. The minor components 2 methylpropionate, 2- and 3-methylbutyrate, phenylacetate, phenylpropionate, and caproate also accumulated, with their relative concentrations varying with the substrate. Succinate was produced in substantial amounts only when corn and alfalfa were fermented; it did not accumulate when cottonseed was the substrate. Samples containing cottonseed were derivatized and subjected to reversed-phase high-performance liquid chromatography, revealing that gossypol concentrations did not change during fermentation. PMID- 11929283 TI - Mineral balance in milk heated using microwave energy. AB - Milk heated to 75 and 85 degrees C in a water bath or in a microwave oven was assayed for changes in salt partitioning after cooling to room temperature. To properly to assess differences and draw valid comparisons, the two heating methods used in the experiment were applied to samples for identical exposure times, and the samples were heated to attain the same final temperatures. Although the soluble Ca and P(i) contents were lower in the heated milk samples, no significant differences in salt partitioning were found between microwave and conventional heating. Ionic calcium levels in the milk samples pasteurized using microwave energy were very close to the levels in the samples heated in a conventional water bath (approximately 90% of the level in the untreated milk samples). The microwave heating-induced changes were completely reversed after storage at 20 degrees C for 24 h. The coagulation properties of the heated milk samples were also examined, and the coagulation time was longer and the curd formation rate slower in the microwave-heated milk than in the raw milk. Still, the experimental results demonstrated that microwave heating was no more detrimental to the milk than conventional heating and could thus be used for pasteurization purposes. PMID- 11929284 TI - Studies on bound (14)C-chlorsulfuron residues in soil. AB - The cause for phytotoxicity of bound residues of chlorsulfuron (2-chloro-N-[[4 methoxy-6-methyl-1,3,5-triazin-2-yl)amino] carbonyl]benzenesulfonamide) to rotational crops is unknown. This study was conducted to determine the formation of nonextractable (bound) residues of chlorsulfuron in soil, and the distribution of bound residues in different organic matter fractions. The results showed that over 150 days, the extractable fraction of (14)C-residues decreased to 25.1% of applied chlorsulfuron, while bound residues concurrently increased to 47.1%. The distribution of (14)C-bound residues in soil organic matter fractions followed an order of humic acid (HA) < humin < fulvic acid (FA). Although the most bound residues were detected in the FA fraction, the amount associated with the humin fraction increased with time. After soil treatment by autoclaving, it was found that bound (14)C-chlorsulfuron residues became available again in the soil. One of the released products was 2-amino-4-hydroxyl-6-methyl-1,3,5-triazine (identified by GC-MS), which is a degradation product of chlorsulfuron. PMID- 11929285 TI - Plant growth inhibitory activity of p-hydroxyacetophenones and tremetones from Chilean endemic Baccharis species and some analogous: a comparative study. AB - Plant growth inhibitory effects of acetophenones 1-6, tremetones 7-12, and MeOH and CH(2)Cl(2) extracts from the aerial parts of Baccharis linnearis, Baccharis magellanica, and Baccharis umbelliformis collected in Chile were assayed as growth inhibitory activity in ranges of 10-500 microM and 0.1-150 ppm, respectively. The effects on seedling growth, germination, and respiration of ryegrass, lettuce, green tomato, and red clover weedy target species were measured. In addition to the inhibitory activity on bleaching of crocin induced by alkoxyl radicals, these compounds also demonstrated scavenging properties toward 2,2-diphenyl-1-picrylhydrazyl in thin-layer chromatography autographic and spectrophotometric assays. In addition, acetophenones and tremetones also showed inhibition of H(+) uptake and oxygen uptake respiration in isolated chloroplasts and mitochondria, respectively. Our results indicate that 1, 4, 7-12, and CH(2)Cl(2) extracts interfere with the dicot preemergence properties, mainly energy metabolism of the seeds at the level of respiration. These compounds appear to have selective effects on the radicle more than shoot growth of dicot seeds. Also, the levels of radicle inhibition obtained with some compounds on Physalis ixocarpa and Trifolium pratense are totally comparable to those of ovatifolin, a known natural growth inhibitor. This behavior might be responsible for its plant growth inhibitory properties and its possible role as an allelopathic agent. PMID- 11929286 TI - Residue uptake and storage responses of Tarocco blood oranges after preharvest thiabendazole spray and postharvest heat treatment. AB - Tarocco blood oranges (Citrus sinensis Linn. Obsek) were subjected to a single preharvest spray with thiabendazole (TBZ) at a concentration of 1% active ingredient (ai) in water and harvested 2 weeks after fungicide application or heated at 37 degrees C for 48 h under saturated humidity after harvest. The two treatments were also combined before cold quarantine (3 weeks at 2 degrees C), subsequent storage (3 weeks at 8 degrees C), and simulated marketing period (SMP) (1 week at 20 degrees C). Fruit not treated with TBZ and unheated were used as controls. The residue levels of TBZ (active ingredient, whole fruit basis) after spray were approximately 6.3 and 5.4 mg x kg(-1) before fruit storage respectively, a level close to the tolerance limit set by the European Community. TBZ showed a high persistence during quarantine, storage, and SMP. TBZ spray significantly reduced the incidence and severity of chilling injury (CI) and decay during the postquarantine period and SMP. Heat treatment (HT) produced beneficial effects in controlling CI, especially during SMP, when applied in combination with TBZ. However, HT remarkably promoted the development of secondary fungal infections such as Phytophthora rots and adversely affected fruit flavor and taste. The occurrence of off-flavor and off-taste was found to be perceptible after heating. PMID- 11929287 TI - Recovery of pigments from Origanum majorana L. by extraction with supercritical carbon dioxide. AB - Extraction of pigments (chlorophylls and carotenoids) from marjoram (Origanum majorana L.) with supercritical carbon dioxide was investigated. The aim of this study was to map the effects of extraction pressure and temperature on the yield of coloring materials by applying a 3(2) full factorial design with three repeated tests in the center of the design. For comparison, laboratory and pilot plant Soxhlet extractions were carried out using ethanol and n-hexane solvents. The compositions of pigments in marjoram extracts were determined by HPLC. Similar amounts of carotenoids, in addition to 40% of chlorophylls and their derivatives, were recovered from the supercritical fluid extraction, in comparison to the ethanol Soxhlet extraction. PMID- 11929288 TI - Identification of betalains from yellow beet (Beta vulgaris L.) and cactus pear [Opuntia ficus-indica (L.) Mill.] by high-performance liquid chromatography electrospray ionization mass spectrometry. AB - Betaxanthins, the yellow-orange water-soluble pigments from yellow beet (Beta vulgaris ssp. vulgaris cv. Bejo Zaden) and cactus pear (Opuntia ficus-indica cv. Gialla) have been investigated using an HPLC system compatible with mass spectrometry. Five novel betaxanthins were found and characterized as the immonium adducts of betalamic acid with serine, gamma-aminobutyric acid, valine, isoleucine, and phenylalanine. To enable concentration of betalain samples, desalting was performed by solid-phase extraction. With this technique, betacyanins could be separated from the betaxanthins using the pH-dependent retention characteristics of red and yellow betalains. The betaxanthin fraction was taken for the preparation of betalamic acid as a precursor for semisynthetic standards. The HPLC method was applied to yellow beet and cactus pear, revealing a more complex betalain profile than described earlier, thus proving its suitability for screening of betaxanthin-containing plants as potential sources for natural food colors. PMID- 11929289 TI - In vitro gastrointestinal digestion study of pomegranate juice phenolic compounds, anthocyanins, and vitamin C. AB - Pomegranate is an important source of bioactive compounds, such as anthocyanins, other phenolic compounds, and ascorbic acid. In the present work an in vitro availability method has been used to assay the influence of the physiological conditions in the stomach and small intestine. This method enables the study of the release of anthocyanins, vitamin C, and total phenols from the pomegranate juice and their transformations during gastrointestinal digestion. Results have shown that pomegranate phenolic compounds are available during the digestion in a quite high amount (29%). Nevertheless, due to pH, anthocyanins are largely transformed into non-red forms and/or degraded (97%), and similar results are obtained for vitamin C (>95% degradation). PMID- 11929290 TI - Study of the organic acids composition of quince (Cydonia oblonga Miller) fruit and jam. AB - The organic acids present in several samples of quince fruit (pulp and peel) and quince jam (homemade and industrially manufactured) were analyzed by HPLC. The sample preparation was simple, involving only extraction with methanol (40 degrees C) and filtration through a Sep-pack C18 cartridge. The chromatographic separation was achieved using an ion exclusion column, Nucleogel Ion 300 OA (300 x 7.7 mm), in conjunction with a column heating device at 30 degrees C. An isocratic elution with H(2)SO(4) 0.01 N as the mobile phase, with a flow rate of 0.1 mL/min, and UV detection at 214 nm were used. These analyses showed that all samples presented a similar profile composed of at least six identified organic acids: citric, ascorbic, malic, quinic, shikimic, and fumaric acids. Several samples also contained oxalic acid. This study suggests that the organic acids levels and ratios may be useful for the determination of percent fruit content of quince jams. The citric acid value can also be used in the differentiation of the type of manufacture of the commercial quince jams (homemade or industrially manufactured). PMID- 11929291 TI - Bacterial removal of quinolizidine alkaloids and other carbon sources from a Lupinus albus aqueous extract. AB - Two Gram-negative bacterial strains capable of using lupanine, the predominant quinolizidine alkaloid in Lupinus albus, as a sole carbon source were isolated from soil in which L. albus and L. luteus had been grown [Santana, F. M. et al. J. Ind. Microbiol. 1996, 17, 110-115]. In the present study, we present results suggesting that these isolates are of potential interest for removing lupanine and other quinolizidine alkaloids (QA) from the effluent resulting from the wet processing of Lupinus seeds, at temperatures within the range 20-34 degrees C. Growth in L. albus aqueous extract was diauxic, with a first period of rapid growth leading to the simultaneous consumption of a significant part of the initial concentration of QA (3 g L(-1), being 2 g L(-1) lupanine) and amino acids (1.5 g L(-1)). This period was followed by a second period of slower growth corresponding to the subsequent partial utilization (25%) of the carbohydrates (initial concentration of 20 g L(-1)) together with further removal of QA and amino acids. Despite the differences detected in the susceptibility of the two strains to lupanine toxicity, in particular at supraoptimal temperatures, and in the efficiency of lupanine catabolism, their performance on L. albus extract did not vary significantly. PMID- 11929293 TI - Oxidation of carbaryl in aqueous solution by membrane anodic fenton treatment. AB - Carbaryl, a commonly used insecticide, was used in this study as a probe to investigate a new Fenton treatment technology, ion exchange membrane anodic Fenton treatment (membrane AFT). It was found that the degradation kinetics of carbaryl by membrane AFT obeys a previously published AFT model quite well. The NaCl (electrolyte) concentration in two half-cells was optimized for two kinds of membrane. Effects of the H(2)O(2)/Fe(2+) ratio and the Fenton reagent delivery rate were also investigated. The treatment efficiency for anion membrane AFT is higher than for salt-bridge AFT under the same operating conditions. Decreasing the delivery rate of Fenton reagents and increasing the treatment temperature also increase the treatment efficiency. The activation energy for carbaryl degradation by anion membrane AFT was estimated to be 14.7 kJ x mol(-1). 1 Naphthol, 1,4-naphthoquinone, and (phthalic acid-O-)yl N-methylcarbamate were detected by GC-MS as the degradation products of carbaryl by Fenton treatment. No decrease in carbaryl degradation rate was found during repeated use (100 times) of the anion exchange membrane. High and stable treatment efficiency can be achieved using an anion exchange membrane rather than a salt-bridge in the AFT system. Because of its effectiveness and convenience, the use of an ion exchange membrane as a substitute for the salt-bridge used in the previous AFT system has brought the AFT technology a major step closer to practical application. PMID- 11929292 TI - Inorganic and organic clays as carriers for controlled release of the herbicide hexazinone. AB - The risk of ground water contamination resulting from rapid leaching of highly soluble pesticides can be minimized through the application of the pesticide adsorbed on a matrix or carrier, which limits the amount of pesticide immediately available for undesirable losses. The use of natural materials for this purpose is of special interest in terms of economy and sustainability. In this work the adsorption of the herbicide hexazinone by two montmorillonites saturated with various inorganic and organic cations was determined and the ability of the two clays displaying the highest adsorption capacities [Fe(3+)-saturated Wyoming montmorillonite, (Fe-SW) and hexadecyltrimethylammonium-saturated Arizona montmorillonite (HDTMA-SA)] to act as carriers for slow release of hexazinone and to reduce herbicide leaching losses was evaluated. Hexazinone formulations based on Fe-SW and HDTMA-SA displayed slow release properties in water and soil/water suspensions, reduced herbicide leaching in soil columns, and maintained herbicidal activity, as compared with the currently available commercial hexazinone formulation (wettable powder). Loosely bound hexazinone-HDTMASA formulations, which led to the slowest breakthrough of hexazinone in soil columns along with the greatest amounts of herbicide released from the clay particles, displayed the most interesting characteristics for their use as slow release formulations and to prevent ground water contamination. PMID- 11929294 TI - Subcritical water extraction to evaluate desorption behavior of organic pesticides in soil. AB - We evaluated the feasibility of extracting organic pesticides in soil using a hot water percolation apparatus at 105 degrees C and 120 kPa pressure. Efficiency of the method was assessed by extracting six selected pesticides (acetochlor, atrazine, diazinon, carbendazim, imidacloprid, and isoproturon) from previously equilibrated soil at 13.6-65.8 mg/kg concentration range. Studies were performed on brown forest soil with clay alluviation (Luvisol). The method developed was compared to the traditional batch equilibrium method in terms of desorbed amount of pesticides from soil and extraction time. Pesticides in the liquid phase from the batch sorption experiment and in the effluent from the hot-water percolation were quantified by high-performance liquid chromatography with UV detection. The results of the percolation experiment are in close correlation with those of the conventional soil testing method. Desorbed quantities by hot-water percolation were 85% acetochlor, 62% atrazine, 65% carbendazim, 44% diazinon, 95% imidacloprid, and 84% isoproturon, whereas using batch equilibrium method 101, 66, 64, 37, 81, and 90% were desorbed, expressed as the percentage of the adsorbed amount of pesticide on soil following equilibration. The average time for hot-water extraction was 3.45 min, in contrast to the 16 h time consumption of the traditional batch method. The effect of temperature on stability of selected compounds was also evaluated using pesticide-spiked sand without soil. Recoveries of analytes ranged between 84.6 and 91.1% with reproducibility of 7.9 10.2%, except for diazinon, for which recovery was 59.4% with 14.4% relative standard deviation since decomposition occurred at elevated temperature. The percolation process has been described by a first-order kinetic equation. The parameters calculated from the equation provide an opportunity to estimate the amount of compound available for desorption, the rate of desorption processes in the studied soil-pesticide-water system, and modeling the leaching process to obtain additional information on the environmental behavior of the examined pesticide. PMID- 11929295 TI - Effect of beta-glycosidase activity of Oenococcus oeni on the glycosylated flavor precursors of Tannat wine during malolactic fermentation. AB - Under traditional wine-making conditions, this work examines the beta-glycosidic activity of Oenococcus oeni on glycosylated aroma compounds of Tannat wines during malolactic fermentation (MLF) by comparing the changes on selected aglycones liberated. MLF diminished the content of all the glycosylated compounds. The level of the free aroma components was slightly modified by the action of the malolactic fermentation so that the cleavage of the glycosidic linkage by the beta-glycosidic activity of O. oeni did not appear to increase significatively the aglycone contents. The consequences of further chemical rearrangements of the algycones under wine conditions were explored using synthesized glycoconjugates on synthetic medium. Bacteria could also be responsible for the cleavage of aroma glycosylated compounds, being the aglycone adsorbed on polysaccharides or peptidoglycans and was released into the external medium. This hypothesis was studied through the evaluation of a stable arrangement of aroma compounds with polysaccharides produced by lactic acid bacteria. A possible retaining of free-made aroma compounds into the whole cells of O. oeni was also investigated through cell culture analysis. Through the results obtained, we assume stable linkage of aroma compounds with bacterial polysaccharides. PMID- 11929296 TI - Generation of roasted notes based on 2-acetyl-2-thiazoline and its precursor, 2 (1-hydroxyethyl)-4,5-dihydrothiazole, by combined bio and thermal approaches. AB - Roasted notes contribute to the flavor of thermally processed foods such as meat and bread. 2-Acetyl-2-thiazoline is one of the key volatile compounds responsible for the roasted and popcorn-like aroma character. We report here on the biogeneration of flavoring preparations with intense roasted notes, which are characterized by a high content of 2-acetyl-2-thiazoline. These flavoring preparations were obtained by fermentation of cysteamine, ethyl-L-lactate, and D glucose with baker's yeast. The precursor of 2-acetyl-2-thiazoline, 2-(1 hydroxyethyl)-4,5-dihydrothiazole, was prepared under mild conditions by microbial reduction of the carbonyl group of 2-acetyl-2-thiazoline using baker's yeast as biocatalyst. The addition of 2-(1-hydroxyethyl)-4,5-dihydrothiazole as aroma precursor to pizza dough resulted in an increase of the roasted note. PMID- 11929297 TI - Degradation of the coffee flavor compound furfuryl mercaptan in model Fenton-type reaction systems. AB - The stability of the coffee flavor compound furfuryl mercaptan has been investigated in aqueous solutions under Fenton-type reaction conditions. The impact of hydrogen peroxide, iron, ascorbic acid, and ethylenediaminetetraacetic acid was studied in various combinations of reagents and temperature. Furfuryl mercaptan reacts readily under Fenton-type reaction conditions, leading to up to 90% degradation within 1 h at 37 degrees C. The losses were lower when one or more of the reagents was omitted or the temperature decreased to 22 degrees C. Volatile reaction products identified were mainly dimers of furfuryl mercaptan, difurfuryl disulfide being the major compound. In addition, a large number of nonvolatile compounds was observed with molecular masses in the range of 92-510 Da. The formation of hydroxyl and carbon-centered radicals was indicated by electron paramagnetic resonance spectra using alpha-(4-pyridyl-1-oxide)-N-tert butylnitrone or 5-(diethoxyphosphoryl)-5-methyl-1-pyrroline-N-oxide as spin traps. Whereas *OH was generated by Fenton-type reactions, the C-centered radical is probably a secondary product of the reaction of *OH with various organic molecules, the reaction with furfuryl mercaptan appearing to be the most important. No evidence for S-centered radicals was seen in the spin-trapping experiments, but a sulfur-containing radical was detected when measurements were made at 77 K in the absence of spin traps. PMID- 11929299 TI - Effects of carnosine on volatile generation from Maillard reaction of ribose and cysteine. AB - Carnosine occurs naturally in meat and meat products in significant quantity, and it possesses strong antioxidant activity that inhibits lipid oxidation and enhances shelf life. In this study, the effects of carnosine on thermal flavor generation were investigated using the model system of cysteine and ribose, which was heated to the roasting temperature of 180 degrees C for 2 h at pH 5 and pH 8.5. The results indicated that carnosine affected volatile formation in a complex manner. Volatiles identified from the liquid phase of the reaction systems of ribose and cysteine showed that the sulfur-containing compounds such as thiophenes, thiazoles, and polysulfides were the most abundant compounds. The addition of carnosine into the reaction mixtures in general caused a reduction in contents of thiophenes and some important meaty flavor compounds such as 2-methyl 3-furanthiol, 2-furfurylthiol, and their associated dimers. On the other hand, it facilitated the generation of several important nitrogen-containing volatiles such as pyrazine, methylpyrazine, 2,6-dimethylpyrazine, and other alkyl pyrazines and thiazoles, which are known to elicit roasty and nutty flavor notes. The results suggested that carnosine acts as a nitrogenous source to facilitate the formation of nitrogen-containing compounds, possibly by degradation to form ammonia. PMID- 11929298 TI - Influence of lipid fraction, emulsifier fraction, and mean particle diameter of oil-in-water emulsions on the release of 20 aroma compounds. AB - The influence of compositional and structural properties of oil-in-water emulsions on aroma release was examined under mouth conditions. The lipid (0.40 and 0.65) and emulsifier fractions (0.007, 0.010, and 0.014) were varied, as well as the mean particle diameter of the dispersed phase (0.60, 0.73, 0.85, and 1.10 microm). Aroma compounds were isolated in a model mouth system and quantified by gas chromatography-mass spectrometry. Studies were carried out to separate effects on the thermodynamic and the kinetic components of aroma release using equilibrium headspace analysis to distinguish the thermodynamic component. The lipid phase of the emulsions was composed of sunflower oil and the emulsifier phase was Tween 20. The release of 20 aroma compounds was evaluated; the compounds included alcohols (1-propanol, 1-butanol, 3-methyl-1-butanol, 2 pentanol, 1-hexanol, and 2-nonanol), ketones (diacetyl, 2-butanone, 2-heptanone, 2-octanone, and 2-decanone), esters (ethyl acetate, propyl acetate, butyl acetate, and ethyl butyrate), aldehydes (hexanal, heptanal, and octanal), a terpene (alpha-pinene), and a sulfur compound (dimethyl sulfide). Decrease in lipid fraction and emulsifier fraction, as well as increase in particle diameter, increased aroma release under mouth conditions. Differences between groups of compounds and between compounds of homologous series with varying chain lengths were found. Changes in particle diameter had a considerable effect on the thermodynamic component of aroma release, whereas hardly any influence of the lipid fraction and emulsifier fraction was observed. Lipid fraction, emulsifier fraction, and particle diameter affected the kinetic component of aroma release, which could partially be attributed to changes in viscosity. PMID- 11929300 TI - Physicochemical and sensory characteristic changes in fortified peanut spreads after 3 months of storage at different temperatures. AB - Three amino acid-balanced, vitamin- and mineral-fortified peanut spreads were stored at 4, 23, and 40 degrees C for 3 months. These were 69.6% peanut/19% soybean-40.5% fat, 61.9% peanut/19% soybean-44.5% fat, and 74.1% peanut/14% nonfat dry milk (NFDM)-40% fat. The peanut spreads were fortified with vitamin A, thiamin, riboflavin, vitamin B(6), vitamin C, calcium, and iron to provide 33.3% of the Recommended Dietary Allowances for children. Water-soluble vitamins were quite stable in deaerated peanut spreads. The antioxidant activity of phytochemicals in soybean prevented vitamin A degradation in soy-containing spreads, whereas the NFDM spread lost 70% of the initial content. Instron analysis detected major changes in texture when peanut spreads were stored at 40 degrees C, suggesting that the polymorphic form of lipid transformed and the proper crystallization of stabilizer was destroyed. Panelists did not detect the texture changes in peanut spreads stored at different temperatures. At 40 degrees C, the primary deteriorative changes in sensory quality were increased browning and the development of "soybean" and "oxidized" flavors as well as decreased "roasted peanutty" flavor. PMID- 11929301 TI - Use of gas chromatography-olfactometry to identify key odorant compounds in dark chocolate. Comparison of samples before and after conching. AB - After vacuum distillation and liquid-liquid extraction, the volatile fractions of dark chocolates were analyzed by gas chromatography-olfactometry and gas chromatography-mass spectrometry. Aroma extract dilution analysis revealed the presence of 33 potent odorants in the neutral/basic fraction. Three of these had a strong chocolate flavor: 2-methylpropanal, 2-methylbutanal, and 3 methylbutanal. Many others were characterized by cocoa/praline flavored/nutty/coffee notes: 2,3-dimethylpyrazine, trimethylpyrazine, tetramethylpyrazine, 3(or 2),5-dimethyl-2(or 3)-ethylpyrazine, 3,5(or 6)-diethyl 2-methylpyrazine, and furfurylpyrrole. Comparisons carried out before and after conching indicate that although no new key odorant is synthesized during the heating process, levels of 2-phenyl-5-methyl-2-hexenal, Furaneol, and branched pyrazines are significantly increased while most Strecker aldehydes are lost by evaporation. PMID- 11929302 TI - Effects of natural phenolic compounds on the antioxidant activity of lactoferrin in liposomes and oil-in-water emulsions. AB - The effect of natural phenolic compounds on the antioxidant and prooxidant activity of lactoferrin was studied in liposomes and oil-in-water emulsions containing iron. The antioxidants tested with lactoferrin were alpha-tocopherol, ferulic acid, coumaric acid, tyrosol, and natural phenolic extracts obtained from three different extra-virgin olive oils and olive mill wastewater. The natural extracts of olive oils and mill wastewaters were composed mainly of polyphenols and simple phenolics, respectively. Lipid oxidation at 30 degrees C was determined by the formation of hydroperoxides and fluorescent compounds resulting from oxidized lipid interactions. All phenolic compounds showed synergistic properties in reinforcing the antioxidant activity of lactoferrin in lipid systems containing iron. The highest synergistic effects were observed for the phenolic extracts rich in polyphenols of extra-virgin olive oils and lactoferrin. This synergistic effect was higher in liposomes than in emulsions. PMID- 11929303 TI - Antitumor activity of compounds isolated from leaves of Eriobotrya japonica. AB - In a search for possible antitumor agents from natural sources, megastigmane glycosides and polyphenolic constituents isolated from the leaves of Eriobotrya japonica (Rosaceae) were found to inhibit the 12-O-tetradecanoylphorbol-13 acetate (TPA)-induced activation of Epstein-Barr virus early antigen in Raji cells. Roseoside and procyanidin B-2 were among the active compounds found in an in vitro assay; these compounds were further assessed for antitumor activity in vivo in a two-stage carcinogenesis assay on mouse skin. Roseoside significantly delayed carcinogenesis induced by peroxynitrite (initiator) and TPA (promoter), and its potency was comparable to that of a green tea polyphenol, (-) epigallocatechin 3-O-gallate, in the same assay. PMID- 11929304 TI - Structural characterization of the metabolites of hydroxytyrosol, the principal phenolic component in olive oil, in rats. AB - Hydroxytyrosol is quantitatively and qualitatively the principal phenolic antioxidant in olive oil. Recently it was shown that hydroxytyrosol and five metabolites were excreted in urine when hydroxytyrosol was dosed intravenously or orally in an olive oil solution to rats. The conclusive identification of three metabolites of hydroxytyrosol by MS/MS as a monosulfate conjugate, a 3-O glucuronide conjugate, and 4-hydroxy-3-methoxyphenylacetic acid (homovanillic acid) has been established in this investigation. The structural configurations of the glucuronide conjugate and 4-hydroxy-3-methoxyphenylacetic acid were confirmed by (1)H NMR. The radical scavenging potencies of homovanillic acid, homovanillic alcohol, hydroxytyrosol, and the metabolites were examined with the radical 2,2-diphenyl-1-picrylhydrazyl. These studies showed them to be potent antioxidants with SC(50) values of 14.8 and 11.4 microM for homovanillic acid and homovanillic alcohol, respectively. The 3-O-glucuronide conjugate was more potent than hydroxytyrosol, with an SC(50) of 2.3 in comparison to 11.0 microM, and the monosulfate conjugate was almost devoid of radical scavenging activity. PMID- 11929305 TI - Activation of polyphenol oxidase in extracts of bran from several wheat (Triticum aestivum) cultivars using organic solvents, detergents, and chaotropes. AB - Polyphenol oxidase (PPO), known to induce browning in wheat-based products, has been shown to be activatable in wheat (Triticum aestivum) bran extracts by chemical compounds. The activity in the extracts could be increased to varying degrees with acetone, methanol, ethanol, 2-propanol, and n-butanol as additives in the extraction buffer. The most potent alcoholic activator was n-butanol (about a 3-fold increase), followed by 2-propanol and ethanol, whereas methanol had the least effect. Ionic detergents in the extraction buffer were also good activators, with sodium dodecyl sulfate (SDS) being more potent (3-fold increase) than cetyltrimethylammonium bromide (CTAB) that had only half as much effect, whereas the nonionic detergent, Triton X-114, was ineffective. The chaotropes, urea and guanidine x HCl (GND), were the most potent activators of all, increasing the activity over 4-fold. Of the two chaotropes, GND was more effective at lower concentrations (<6 M) than urea. However, the enzyme activity lessened at a higher concentration of GND (6 M), while there was a further increase in the activity with 6 M urea treatment. The activity lessened with higher concentration of GND presumably as a result of extensive denaturation of the enzyme, as GND is known to be a more potent denaturant than urea. It is hypothesized that in wheat PPO exists in an inactive form which may be activated by the presence of activators, hitherto unknown, similar in effect to that elicited by the chemical denaturants in this study. PMID- 11929306 TI - Protective activity of green tea against free radical- and glucose-mediated protein damage. AB - Protein oxidation and glycation are posttranslational modifications that are implicated in the pathological development of many age-related disease processes. This study investigated the effects of green tea extract, and a green tea tannin mixture and its components, on protein damage induced by 2,2'-azobis(2 amidinopropane) dihydrochloride (a free radical generator) and glucose in in vitro assay systems. We found that green tea extract can effectively protect against protein damage, and showed that its action is mainly due to tannin. In addition, it was shown that the chemical structures of tannin components are also involved in this activity, suggesting that the presence of the gallate group at the 3 position plays the most important role in the protective activity against protein oxidation and glycation, and that there is also a contribution by the hydroxyl group at the 5' position in the B ring and the sterical structure. These findings demonstrate the mechanisms of the usefulness of green tea in protein oxidation- and glycation-associated diseases. PMID- 11929307 TI - Arabinoxylan-lipid-based edible films and coatings. 3. Influence of drying temperature on film structure and functional properties. AB - This work is a contribution to better knowledge of the influence of the structure of films obtained from emulsions based on arabinoxylans, hydrogenated palm kernel oil, and emulsifiers on their functional properties. The sucrose esters (emulsifiers) have a great effect on the stabilization of the emulsified film structure containing arabinoxylans and hydrogenated palm kernel oil. The structure and stability of the emulsion during drying strongly affect barrier and mechanical properties of films. The higher are creaming and coalescence phenomena in films, the lower is the water vapor permeability. Emulsion destabilization is favored by high drying temperature and tends to give films having a "bilayer like" structure, which tends to improve the functional properties of arabinoxylans-based edible films. PMID- 11929308 TI - Enzymatic synthesis of [3'-O-methyl-(3)H]malvidin-3-glucoside from petunidin-3 glucoside. AB - Malvidin-3-glucoside has been labeled by enzymatic synthesis in a single-step experiment. Catechol-O-methyl transferase catalyzed the B-ring methylation of petunidin-3-glucoside, and S-Adenosyl-L-[methyl-(3)H] methionine was the methyl donor. Solid phase extraction and preparative high-performance liquid chromatography were necessary to separate [3'-O-methyl-(3)H]malvidin-3-glucoside from an isomer and the starting material. The specific activity was 2.2 Ci mmol( 1), and the yield of incorporation was 1.1%. A possible application of the labeled material is the study of anthocyanin reactions in complex mixtures such as red wine where products are difficult to isolate and analyze. PMID- 11929309 TI - Phenolic compounds and antioxidant capacity of Georgia-grown blueberries and blackberries. AB - Blueberries and blackberries grown at various locations in Georgia were collected and analyzed for flavonoids, total anthocyanins, total polyphenols, and Trolox equivalent antioxidant capacity (TEAC). Each sample was analyzed for phenolic acids (gallic acid, p-hydroxybenzoic acid, caffeic acid, ferulic acid, and ellagic acid) and flavonoids (catechin, epicatechin, myricetin, quercetin, and kaempferol). A high-performance liquid chromatographic (HPLC) method with photodiode array detection was used for analysis. Compounds were analyzed as aglycons after acid hydrolysis with 1.2 M HCl. Identification of each compound was based on retention time and UV spectra by comparison with pure commercial standards. Phenolic acids ranged from 0.19 to 258.90 mg/100 g fresh weight (FW), and flavonoids ranged from 2.50 to 387.48 mg/100 g FW. Total polyphenols ranged from 261.95 to 929.62 mg/100 g FW, and total anthocyanins ranged from 12.70 to 197.34 mg/100 g FW. TEAC values varied from 8.11 to a maximum of 38.29 microM/g FW. A linear relationship was observed between TEAC values and total polyphenols or total anthocyanins. The data indicate that blueberries and blackberries are rich sources of antioxidants. PMID- 11929311 TI - Classification and class-modeling of "Riviera Ligure" extra-virgin olive oil using chemical-physical parameters. AB - The Protected Designation of Origin (PDO) "Riviera Ligure" for extra-virgin olive oils from Liguria specifies three additional geographical mentions corresponding to three different geographical areas. To obtain a complete characterization of this typical Italian product, 217 samples of olive oils produced in this North Italian region during 1998/99 and 1999/2000 were analyzed. For each sample 31 variables were determined by chemical-physical analyses, and the data were subjected to a multivariate statistical analysis. For the 1998/99 crop, characterized by favorable climatic conditions, class-models of the three geographical areas were obtained with good predictive ability, also considering the influence of the month of olive harvesting. The oil samples from the 1999/2000 crop were similarly studied, but bad climatic conditions and a widespread Dacus oleae infestation leveled out the peculiar features of the oils produced in the three areas. PMID- 11929310 TI - Characterization of "lettucine", a serine-like protease from Lactuca sativa leaves, as a novel enzyme for milk clotting. AB - In this work we focused on the characterization of a novel plant rennet purified from lettuce leaves (Lactuca sativa L. cv Romana). The lettuce protease, lettucine, showed trypsin-like, SV8-like, and caseinolytic activities. Although the enzyme did not recognize peptides having hydrophobic amino acid residues in the P(1) position of the target bond, it did show milk-clotting activity, suggesting that different bonds rather than the Phe(105)-Met(106) of the kappa casein might be cleaved, still inducing milk-clotting. The enzyme exhibited proteolytic activity toward alpha-casein, beta-casein, kappa-casein, and milks with different fat contents, with the highest activity observed with partially skimmed milk, total casein, and alpha- and kappa-casein. SDS-PAGE studies showed that lettucine cleaved alpha-casein, beta-casein, and kappa-casein. In particular, we showed that alpha-casein breakdown occurred even though total casein or milks were supplied, suggesting that the lettuce enzyme is able to operate a significant disorganization of the casein's micellar structure. Moreover, the proteolytic activity of the enzyme analyzed under various technological parameters, such as temperature and pH, indicated that the lettuce enzyme is highly consistent with the milk-clotting process. PMID- 11929312 TI - Using transgenic poplars to elucidate the relationship between the structure and the thermal properties of lignins. AB - In an attempt to draw relationships between the molecular structure and the thermal behavior of lignins, thermomechanical analyses were run on six milled wood and enzyme poplar lignin fractions prepared from genetically modified and control woods. All the lignin samples displayed similar thermal profiles with a clear inflection point assigned to the glass transition point. The temperature (T(g)) at which this transition occurs showed large variations from 170 to 190 degrees C, depending both on the genetic modification and on the age of the tree. These variations were found to be closely related to the condensation degree of lignins evaluated by thioacidolysis. PMID- 11929313 TI - Antioxidative activity and safety of the 50 ethanolic extract from red bean fermented by Bacillus subtilis IMR-NK1. AB - This study aimed at evaluating the antioxidative activities and the safety of 50% ethanolic extract from red bean fermented by Bacillus subtillis IMR-NK1. The antioxidative activities, including alpha,alpha-diphenyl-beta-picryl-hydrazyl (DPPH) radicals scavenging effects, Fe(2+)-chelating ability, and reducing power, were studied in vitro. It was found that the antioxidative activity increased with the concentrations of the extract to a certain extent and then leveled off as the concentration further increased. As compared to the commercial antioxidants, the fermented red bean extract showed less scavenging effect on the DPPH radical and reducing power than alpha-tocopherol and BHT, but better Fe(2+) chelating ability. No mutagenicity or toxicity effect toward all tester strains was found in the 50% ethanolic extract of fermented red bean by means of the Ames test. The results suggested that the 50% ethanolic extract was safe in genotoxicity. PMID- 11929314 TI - Antioxidative phenolic compounds isolated from almond skins (Prunus amygdalus Batsch). AB - Nine phenolic compounds were isolated from the ethyl acetate and n-butanol fractions of almond (Prunus amygdalus) skins. On the basis of NMR data, MS data, and comparison with the literature, these compounds were identified as 3'-O methylquercetin 3-O-beta-D-glucopyranoside (1); 3'-O-methylquercetin 3-O-beta-D galactopyranoside (2); 3'-O-methylquercetin 3-O-alpha-L-rhamnopyranosyl-(1-->6) beta-D-glucopyranoside (3); kaempferol 3-O-alpha-L-rhamnopyranosyl-(1-->6)-beta-D glucopyranoside (4); naringenin 7-O-beta-D-glucopyranoside (5); catechin (6); protocatechuic acid (7); vanillic acid (8); and p-hydroxybenzoic acid (9). All of these compounds have been isolated from almond skins for the first time. 2,2 Diphenyl-1-picrylhydrazyl (DPPH) free radical scavenging activities for compounds 1-9 were determined. Compounds 6 and 7 show very strong DPPH radical scavenging activity. Compounds 1-3, 5, 8, and 9 show strong activity, whereas compound 4 has very weak activity. PMID- 11929315 TI - Comparison of radical scavenging effect, inhibition of microsomal oxygen free radical generation, and serum lipoprotein oxidation of several natural antioxidants. AB - Typical components of the Mediterranean diet, such as olive oil and red wine, contain high concentrations of complex phenols, which have been suggested to have an important antioxidant role. The aim of the present work was to determine the inhibitory potency of compounds such as oleuropein, hydroxytyrosol, and other structurally related compounds, such as gallic acid, toward reactive oxygen species generation and free radical scavenging ability. The potency of these compounds was also examined with respect to protecting in vitro low-density lipoprotein oxidation. These studies indicate that complex phenols, such as hydroxytyrosol, and gallic acid both inhibit free radical generation and act as free radical scavengers. The use of three different approaches to determine antioxidant potency demonstrates that activity in one test does not necessarily correlate with activity in another. It was also demonstrated that the presence of two phenolic groups is not always associated with antioxidant activity. PMID- 11929317 TI - The product label: how pharmacokinetics and pharmacodynamics reach the prescriber. AB - The product label, or package insert, is the 'manual' for the safe and effective use of a drug. Important pharmacokinetic and pharmacodynamic properties of a drug product should appear in the label under specific sections, as required in the Code of Federal Regulations (CFR), using a format and language recommended by the Food and Drug Administration (FDA) in various guidances to the industry. The relevant regulations and guidance documents impacting on how this information is conveyed to the healthcare professional are discussed, with special emphasis on how the new proposed rule will impact upon how information is to be conveyed. With the availability of new clinical pharmacology information not available at the time of approval, package inserts for older drugs should be updated to reflect the new data and recommend the proper dosage regimen, enabling prescribers to optimise drug therapy and minimise possible adverse events. PMID- 11929319 TI - Clinical pharmacokinetics of tirofiban, a nonpeptide glycoprotein IIb/IIIa receptor antagonist: comparison with the monoclonal antibody abciximab. AB - Tirofiban is a nonpeptide tyrosine derivative that antagonises platelet glycoprotein IIb/IIIa (GP IIb/IIIa) receptors. It is one of three GP IIb/IIIa antagonists approved by the US Food and Drug Administration for the treatment of patients with acute coronary syndromes. The clinical effect of tirofiban has been shown in large studies such as PRISM (Platelet Receptor Inhibition for Ischemic Syndrome Management), PRISM-PLUS (PRISM - Patients Limited by Unstable Signs and Symptoms) and RESTORE (Randomised Efficacy Study of Tirofiban for Outcomes and Restenosis). Tirofiban is administered as an intravenous infusion. Volume of distribution ranges from 21 to 87 L, and binding to human plasma proteins is modest at 64%. Metabolism in humans is negligible, and most drug is excreted renally with systemic clearance ranging from 4.8 to 25.8 L/h. Renal function may influence the excretion of tirofiban, but concurrent disease or other drugs generally used in patients with ischaemia seem not to do so. This review updates what is known about the pharmacokinetics of tirofiban in humans, especially in comparison with the monoclonal antibody against the IIb/IIIa receptor, abciximab. PMID- 11929318 TI - Strategies for increasing drug delivery to the brain: focus on brain lymphoma. AB - The blood-brain barrier (BBB) is a gate that controls the influx and efflux of a wide variety of substances and consequently restricts the delivery of drugs into the central nervous system (CNS). Brain tumours may disrupt the function of this barrier locally and nonhomogeneously. Therefore, the delivery of drugs to brain tumours has long been a controversial subject. The current concept is that inadequate drug delivery is a major factor that explains the unsatisfactory response of chemosensitive brain tumours. Various strategies have been devised to circumvent the BBB in order to increase drug delivery to the CNS. The various approaches can be categorised as those that attempt to increase delivery of intravascularly administered drugs, and those that attempt to increase delivery by local drug administration. Strategies that increase delivery of intravascularly injected drugs can manipulate either the drugs or the capillary permeability of the various barriers (BBB or blood-tumour barrier), or may attempt to increase plasma concentration or the fraction of the drug reaching the tumour (high-dose chemotherapy, intra-arterial injection). Neurotoxicity is a major concern with increased penetration of drugs into the CNS or when local delivery is practised. Systemic toxicity remains the limiting factor for most methods that use intravascular delivery. This review evaluates the strategies used to increase drug delivery in view of current knowledge of drug pharmacokinetics and its relevance to clinical studies of chemosensitive brain tumours. The main focus is on primary CNS lymphoma, as it is a chemosensitive brain tumour and its management routinely utilises specialised strategies to enhance drug delivery to the affected CNS compartments. PMID- 11929320 TI - Clinical pharmacokinetics of dorzolamide. AB - Dorzolamide is a carbonic anhydrase inhibitor for topical ophthalmic application. It is used in the treatment of glaucoma to lower the intraocular pressure. After absorption via the cornea and stroma, it inhibits carbonic anhydrase in the ciliary process, which leads to a reduction of aqueous humour production and therefore to the desired therapeutic effect. In the systemic circulation, dorzolamide is bound mainly to carbonic anhydrase in red blood cells. It is slowly metabolised to N-de-ethyldorzolamide, which in turn is also stored in red blood cells. The very slow elimination (half-life >4 months) of both substances takes place via the renal route. However, the inhibition of carbonic anhydrase in red blood cells is moderate in the course of a topical treatment, avoiding systemic adverse effects. This review summarises the pharmacokinetic and pharmacodynamic properties of dorzolamide and its metabolite in eye tissues and in the systemic circulation. PMID- 11929322 TI - Estimation of the absolute bioavailability of rivastigmine in patients with mild to moderate dementia of the Alzheimer's type. AB - OBJECTIVE: To investigate the bioavailability of rivastigmine, an approved therapy for patients with mild to moderate dementia of the Alzheimer's type, at the highest approved single dose of 6 mg. DESIGN AND SETTING: Randomised, two period crossover, single-centre, non-blinded, inpatient study. PATIENTS AND PARTICIPANTS: Eleven patients (five females and six males) with mean age 69.5 years. METHODS: The 6 mg oral dose was compared with a 2 mg intravenous dose of rivastigmine infused over a 1-hour period. Plasma concentrations of rivastigmine and its metabolite NAP 226-90 were measured with a gas chromatographic/mass spectrometric method. RESULTS: Following oral administration of a single 6 mg capsule, rivastigmine is rapidly absorbed with an average time to peak plasma concentration of about 1 hour and an average peak concentration of about 25.6 g/L. By a noncompartmental approach, the absolute bioavailability of the 6 mg oral dose of rivastigmine was 71.7% when compared with a 2mg intravenous infusion normalised for dose. By using a population pharmacokinetic model with Michaelis Menten elimination, absolute bioavailability was estimated at 60.2%. The average terminal elimination half-life of rivastigmine ranged from 1.4 to 1.7 hours for both treatments. Plasma concentrations of the major metabolite, NAP 226-90, formed by the hydrolysis of rivastigmine by cholinesterase are lower than those of the parent compound following oral and intravenous administration. CONCLUSION: A noncompartmental approach and a compartmental approach based on a population pharmacokinetic model with Michaelis-Menten elimination yielded comparable values, 71.7% and 60.2% respectively, for the absolute bioavailability of a single 6 mg oral dose of rivastigmine. Comparison with previous studies confirmed that the oral form of the drug exhibits increased bioavailability with increasing dose, consistent with its nonlinear pharmacokinetics.. PMID- 11929321 TI - Clinical pharmacokinetics and selective pharmacodynamics of new angiotensin converting enzyme inhibitors: an update. AB - The angiotensin converting enzyme (ACE) inhibitors are widely used in the management of essential hypertension, stable chronic heart failure, myocardial infarction (MI) and diabetic nephropathy. There is an increasing number of new agents to add to the nine ACE inhibitors (benazepril, cilazapril, delapril, fosinopril, lisinopril, pentopril, perindopril, quinapril and ramipril) reviewed in this journal in 1990. The pharmacokinetic properties of five newer ACE inhibitors (trandolapril, moexipril, spirapril, temocapril and imidapril) are reviewed in this update. All of these new agents are characterised by having a carboxyl functional groups and requiring hepatic activation to form pharmacologically active metabolites. They achieve peak plasma concentrations at similar times (t(max)) to those of established agents. Three of these agents (trandolapril, moexipril and imidapril) require dosage reductions in patients with renal impairment. Dosage reductions of moexipril and temocapril are recommended for elderly patients, and dosages of moexipril should be lower in patients who are hepatically impaired. Moexipril should be taken 1 hour before meals, whereas other ACE inhibitors can be taken without regard to meals. The pharmacokinetics of warfarin are not altered by concomitant administration with trandolapril or moexipril. Although imidapril and spirapril have no effect on digoxin pharmacokinetics, the area under the concentration-time curve of imidapril and the peak plasma concentration of the active metabolite imidaprilat are decreased when imidapril is given together with digoxin. Although six ACE inhibitors (captopril, enalapril, fosinopril, lisinopril, quinapril and ramipril) have been approved for use in heart failure by the US Food and Drug Administration, an overview of 32 clinical trials of ACE inhibitors in heart failure showed that no significant heterogeneity in mortality was found among enalapril, ramipril, quinapril, captopril, lisinopril, benazepril, perindopril and cilazapril. Initiation of therapy with captopril, ramipril, and trandolapril at least 3 days after an acute MI resulted in all-cause mortality risk reductions of 18 to 27%. Captopril has been shown to have similar morbidity and mortality benefits to those of diuretics and beta-blockers in hypertensive patients. Captopril has been shown to delay the progression of diabetic nephropathy, and enalapril and lisinopril prevent the development of nephropathy in normoalbuminuric patients with diabetes. ACE inhibitors are generally characterised by flat dose-response curves. Lisinopril is the only ACE inhibitor that exhibits a linear dose-response curve. Despite the fact that most ACE inhibitors are recommended for once-daily administration, only fosinopril, ramipril, and trandolapril have trough-to-peak effect ratios in excess of 50%. PMID- 11929323 TI - The underuse of therapy in the secondary prevention of hip fractures. AB - There is strong evidence to indicate that individuals who sustain a hip fracture are at a greater risk of developing another. The management of such patients should include efforts to prevent future fractures, including prescribing medications that have been shown to lower hip fracture risk. Such therapies that are currently available include calcium and vitamin D supplementation, alendronic acid and risedronic acid. In addition, there is epidemiological evidence to indicate that estrogen may also decrease the risk of hip fracture. Parathyroid hormone is another agent that has shown promise in this regard and is likely to be available for clinical use in the near future. However, the rates of utilisation of these therapies among patients with hip fractures are low. It is important to emphasise that secondary prevention of hip fractures should be an integral part of the management of individuals who sustain hip fractures. PMID- 11929324 TI - Identification of suicidal ideation and prevention of suicidal behaviour in the elderly. AB - In almost all industrialised countries, men aged 75 years and older have the highest suicide rate among all age groups. Although in younger age groups suicide attempts are often impulsive and communicative acts, suicide attempts in older people (defined as aged 65 years and older) are often long planned and involve high-lethality methods. These characteristics, in addition to the fact that elderly are more fragile and frequently live alone, more often lead to fatal outcome. In later life, in both sexes, the most common diagnosis in those who attempt or complete suicide is major depression. In contrast to other age groups, comorbidity with substance abuse and personality disorders is less frequent. Physical illness plays an important role in the suicidal behaviour of the elderly: most frequently, depression and illness co-occur; less often, the physical illness or the treating medications are causally related to the depressive symptoms. However, only 2 to 4% of terminally ill elderly commit suicide. In addition to physical illness, complicated or traumatic grief, anxiety, unremitting hopelessness after recovery from a depressive episode and history of previous suicide attempts are risk factors for suicide attempts and completed suicide. During a depressive episode, elderly patients with suicidal ideation have higher levels of anxiety and, during treatment, anxiety decreases the probability of remission and recovery. As well as overt suicide attempts, indirect self-destructive behaviours, which often lead to premature death, are common, especially in residents of nursing homes, where more immediate means to commit suicide are restricted. Although we do not have randomised trials of treatment, studies suggest that antidepressant treatment may decrease suicide risk. Prevention and treatment trials are underway to detect the effectiveness of improved treatment of depression by primary care physicians as a means of reducing the prevalence of depressive symptoms, hopelessness and suicidal ideation. PMID- 11929325 TI - Pharmacokinetic considerations of oral chemotherapy in elderly patients with cancer. AB - Persons over the age of 65 years are the fastest growing segment of the US population. In the next 30 years they will comprise over 20% of the population. Fifty per cent of all cancers occur in this age group and therefore there will be an expected rise in the total cancer burden. There has been an increasing trend over the past 20 years toward the use of oral chemotherapy. This change has been encouraged by the need to decrease the costs of chemotherapy administration, patient preferences and quality of life issues. Factors that must be considered with oral chemotherapy administration include limitations of saturability of absorption, patient compliance and pharmacokinetic/pharmacodynamic changes which occur in elderly patients. Interpatient variability and drug metabolism, particularly age-related changes in drug metabolism are being studied. The cytochrome P450 system has been intensively studied because of its importance with regard to chemotherapeutic drugs. This article reviews these issues and provides details regarding specific drugs including temozolomide, thalidomide, topotecan, the fluoropyrimidines, etoposide, hydroxycarbamide (hydroxyurea), tamoxifen, and alkylating drugs. Complementary and alternative therapies are also discussed. PMID- 11929327 TI - Rationale for the use of aldosterone antagonists in congestive heart failure. AB - Traditionally, the role of aldosterone in heart failure was thought to be a result of its effects on epithelial cells where it induces sodium reabsorption and potassium excretion with subsequent haemodynamic effects from intravascular volume expansion. On this basis, spironolactone, a non-selective aldosterone antagonist, has been used for the treatment of congestive heart failure to block aldosterone-mediated effects in epithelial cells. The Randomized Aldactone Evaluation Study (RALES), in which spironolactone was added to existing therapy in patients with heart failure, showed a significant reduction in morbidity and mortality. These results suggest that the role of aldosterone in the pathophysiology of cardiovascular disease may be more complex than previously recognised. There now is extensive experimental and growing clinical evidence for an important physiological role for aldosterone in the pathology of cardiac and renal disease. Classical effects of aldosterone are mediated via its nuclear receptor. Novel non-epithelial effects of aldosterone are mediated via a second messenger system, which involves activation of the sodium/hydrogen antiporter. These effects of aldosterone have been demonstrated in the kidney, vascular smooth muscle cell and leukocytes, and in the regulation of rapid corticotropin suppression. It has been hypothesised that cardiac damage induced by aldosterone is independent of the presence of hypertension. In support of this, experimental evidence demonstrates that cardiovascular damage induced by aldosterone can be prevented by administration of a selective mineralocorticoid receptor antagonist. These findings suggest the dissociation between cardiovascular lesions and high blood pressure, and highlight the importance of aldosterone in the pathological changes. PMID- 11929328 TI - New prospects for the treatment of lysosomal storage diseases. AB - Although individually rare, lysosomal storage disorders constitute a significant burden on society. To date, enzyme replacement therapy (ERT) has been the most successful therapeutic approach for lysosomal storage disorders. ERT reverses systemic manifestations of Gaucher disease but does not effectively treat the neurological complications. Recently, ERT produced a reduction of severe neuropathic pain, stabilisation of renal disease, and improved vascular function and structure in short-term, placebo-controlled trials in patients with Fabry's disease. Long-term studies are necessary to evaluate the full potential of ERT in this disease. In patients with Pompe disease, a fatal cardiac and skeletal muscle disorder, ERT improved cardiac function and structure, and increased overall muscle strength. It has already increased survival in a small number of affected infants. ERT also decreased liver and spleen size, joint mobility and quality of life in patients with mucopolysaccharidosis type I, but when the therapeutic protein is administered intravenously, it is unlikely to modify the neurological outcome in this or in other similar disorders. Bone marrow transplantation continues to be effective in Gaucher disease, in some forms of mucopolysaccharidosis and in mild forms of Krabbe disease, but it has high morbidity and mortality that limits its use in lysosomal storage disorders. Drugs that slow the rate of formation of accumulating glycolipids are being developed and one of them, OGT-918 (N-butyldeoxynojirimycin), is showing promise in patients with Gaucher disease. Gene therapy for lysosomal storage disorders holds promise as a replacement for the other therapies described here but requires much more development before clinical efficacy trials. PMID- 11929329 TI - Substance misuse in patients with schizophrenia: epidemiology and management. AB - Substance misuse in individuals with schizophrenia is very common, especially in young men, in communities where use is frequent and in people receiving inpatient treatment. Problematic use occurs at very low intake levels, so that most affected people are not physically dependent (with the exception of nicotine). People with schizophrenia and substance misuse have poorer symptomatic and functional outcomes than those with schizophrenia alone. Unless there is routine screening, substance misuse is often missed in assessments. Service systems tend to be separated, with poor inter-communication, and affected patients are often excluded from services because of their comorbidity. However, effective management of these disorders requires a fully integrated approach because of the close inter-relationship of the disorders. Use of atypical antipsychotics may be especially important in this population because of growing evidence (especially on clozapine and risperidone) that nicotine smoking, alcohol misuse and possibly some other substance misuse is reduced. Several pharmacotherapies for substance misuse can be used safely in people with schizophrenia, but the evidence base is small and guidelines for their use are necessarily derived from experience in the general population. PMID- 11929330 TI - Fetal pharmacotherapy. AB - Rapid progress has recently been encountered in pharmacologically treating the unborn baby. This unique area of drug therapy raises new methodological and ethical questions. This article is a systematic review of known modalities of fetal pharmacotherapy, and aims to highlight essential principles, difficulties and controversies in fetal pharmacotherapy. Unique pharmacokinetic features of pregnancy, the placenta and the fetus govern maternal-to-fetal drug transfer. Ethically, it is important that the mother and family are appropriately informed about the evidence in favour of specific fetal therapy, its risks and alternatives. Antenatal use of corticosteroids for lung maturation is an example of adequate methodology, leading to clear results. In contrast, the initial hopes in antenatal use of phenobarbital were based on less than optimal methodology. Folic acid for the prevention of neural tube defects is the first instance of fetal therapy that has led to the prevention of a major malformation. Serious infections, such as HIV, Group B streptococcus and toxoplasmosis highlight the need for controlled, randomised studies to prevent fetal infection. With scores of new modalities of fetal therapy likely to be introduced in the next few years, it will be important to adhere to the best possible methodology and execution, in order to address optimally the needs of the fetus. PMID- 11929331 TI - Emerging therapies in the pharmacological treatment of Parkinson's disease. AB - The pharmacological management of Parkinson's disease is a complex and dynamic task; there is no one 'right' strategy indicating which drugs should be used at a particular stage of the disease. There are now many different drugs belonging to several classes that may be effective, and there are still differences of opinion among leading clinicians about the best course of treatment. This review focuses on drug therapy for the motor impairment in Parkinson's disease. Current and future research directions are summarised by taking inventory of recent and innovative areas of development in the field, representing each category with at least one of its featured treatments. The main research efforts are being directed towards delaying the use of levodopa or finding therapies to be used as adjunct to it, in order to postpone motor complications and, in particular, dyskinesias. One of the recent trends is early employment of dopamine agonists. Additional efforts are being directed towards protecting and restoring dopamine neurons. Novel therapies acting on non-dopaminergic systems are also being researched. PMID- 11929333 TI - Topical pimecrolimus: a review of its clinical potential in the management of atopic dermatitis. AB - Pimecrolimus (SDZ ASM 981), an ascomycin derivative, is a nonsteroid, has anti inflammatory activity, and has demonstrated efficacy in reducing symptoms of atopic dermatitis in adult and paediatric patients when applied topically. Compared with vehicle, topical pimecrolimus 1.0% cream was significantly more effective at reducing symptoms of atopic dermatitis, as measured by the Eczema Area and Severity Index (EASI), in infants aged 3 to 23 months, children aged 2 to 17 years and adults. The median reductions from baseline in the total EASI score in adults after treatment with pimecrolimus 1.0% or corresponding vehicle twice daily for 3 weeks were 47 and 0%, respectively. In infants and children, treatment with pimecrolimus 1.0% twice daily for 6 weeks resulted in significant decreases in mean EASI scores compared with vehicle. The severity of pruritus was significantly reduced in patients of all age groups after topical treatment with pimecrolimus 1.0% cream. Compared with vehicle, the incidence of eczematous flares was also reduced by intermittent long-term use of topical pimecrolimus 1.0% in adults, children and infants. Sixty percent of children treated with pimecrolimus for 1 year completed the first 6 months of treatment without experiencing a flare, compared with 35% of patients who received vehicle. Furthermore, the use of topical corticosteroids for the treatment of uncontrolled flares in adults, children and infants was lower in the pimecrolimus groups than in the vehicle groups. Topical pimecrolimus 1.0% cream is well tolerated in atopic dermatitis patients of all age groups. There were no clinically relevant systemic adverse events reported from any of the studies in patients with atopic dermatitis. The most frequently reported adverse events pertained to application site reactions, such as burning and a feeling of warmth. In conclusion, topical pimecrolimus 1.0% cream has shown efficacy in the treatment of mild to moderate atopic dermatitis in infants, children and adults. Although tolerability data concerning infants and children have not yet been published in full, the drug appears to be well tolerated in all age groups, and there have been no reports of clinically relevant systemic adverse events. Furthermore, pimecrolimus 1.0% cream has shown no potential for skin atrophy, a problem commonly associated with treatment with topical corticosteroids. Pimecrolimus 1.0% cream provides a promising and well tolerated treatment option in the management of infants, children and adults with mild to moderate atopic dermatitis. PMID- 11929332 TI - Candesartan cilexetil plus hydrochlorothiazide combination: a review of its use in hypertension. AB - The combination of candesartan cilexetil [an angiotensin II type 1 (AT(1)) receptor antagonist] plus hydrochlorothiazide (a thiazide diuretic), has been used in the treatment of patients with hypertension. The blood pressure (BP) lowering effect of various doses of this combination, administered orally once a day for 4 to 52 weeks, has been demonstrated in clinical trials. These studies showed that combinations of candesartan cilexetil 4 to 16 mg with hydrochlorothiazide 12.5 or 25 mg induced significant reductions reductions in systolic (S) BP and diastolic (D) BP from baseline in patients with mild to severe hypertension. Data from clinical trials indicated that reductions in BP induced by candesartan cilexetil 4 to 32 mg/hydrochlorothiazide 12.5 mg combinations were significantly greater than those observed after monotherapy with either drug. Treatment for 8 weeks with candesartan cilexetil 16 mg/hydrochlorothiazide 12.5 mg or candesartan cilexetil 16 mg induced SBP/DBP reductions of 12.0/7.5 mm Hg and 7.5/5.5mm Hg, respectively (p < 0.05 both comparisons). Moreover, data from a randomised, double-blind, placebo-controlled, dose-finding study in 1038 patients with mild to moderate hypertension showed that the greatest reductions in SBP/DBP were achieved by candesartan cilexetil 16 mg/hydrochlorothiazide 12.5 mg. Significant differences in BP reduction in favour of the combination were observed when hypertensive patients were given candesartan cilexetil 4 or 8 mg/hydrochlorothiazide 12.5 mg or hydrochlorothiazide monotherapy for 8 weeks. Additionally, greater efficacy of the combination compared to monotherapy with either drug was demonstrated by response rates to treatment. Moreover, a fixed combination of candesartan cilexetil 16 mg/hydrochlorothiazide 12.5 mg demonstrated a greater antihypertensive effect than losartan 50 mg/hydrochlorothiazide 12.5 mg in two clinical trials. Candesartan cilexetil 8 mg/hydrochlorothiazide 12.5 mg showed a similar antihypertensive effect compared with that of combined lisinopril 10 mg/hydrochlorothiazide 12.5 mg. Candesartan cilexetil/hydrochlorothiazide combination was well tolerated in patients with hypertension. Combined data from placebo-controlled trials showed that most adverse events were uncommon and not serious. Patients receiving combination therapy exhibited, among other adverse events, headache (3.2 vs 5.5% for candesartan cilexetil/hydrochlorothiazide and placebo, respectively), back pain (3.0 vs 2.4%), dizziness (2.6 vs 1.2%) and respiratory infection (2.5 vs 1.4%). Moreover, 3.3 and 2.7% of patients receiving candesartan cilexetil/hydrochlorothiazide or placebo, respectively, discontinued treatment because of adverse events. CONCLUSION: The combination of candesartan cilexetil and hydrochlorothiazide (AT(1)-receptor antagonist and thiazide diuretic, respectively) is an effective treatment for patients with hypertension. Data from randomised, double-blind, placebo-controlled clinical trials showed that this combination is significantly more efficacious than either agent alone. Moreover, the combination of these two agents showed an excellent adverse event profile. Current data support the use of this combination as an alternative when monotherapy with either agent is not effective, and there are no compelling or specific indications for other drugs. However, data from large clinical trials, evaluating morbidity and mortality outcomes, are needed to determine the precise role of candesartan cilexetil/hydrochlorothiazide combination in the treatment of patients with hypertension. PMID- 11929334 TI - Bivalirudin: a review of its potential place in the management of acute coronary syndromes. AB - Bivalirudin, a synthetic analogue of hirudin, is a specific and reversible inhibitor of thrombin which binds directly with both fluid-phase and clot-bound thrombin. In patients with unstable angina undergoing percutaneous transluminal coronary angioplasty (PTCA), results from a large well designed study and its reanalysis (n = 4312) indicate that bivalirudin is more effective than heparin in the prevention of ischaemic complications for up to 90 days after the start of treatment. In addition, among patients undergoing PTCA for post myocardial infarction (MI) bivalirudin may be more effective than heparin in preventing ischaemic complications for up to 180 days after treatment was started. Data from dose-finding studies indicate bivalirudin has potential in the treatment of patients with unstable angina not undergoing percutaneous coronary intervention (PCI); however, well designed comparative studies are needed before firm conclusions can be made. Among patients with acute ST elevation MI, randomised trials have demonstrated bivalirudin to be significantly more effective than heparin in improving early patency in patients receiving thrombolytic therapy with streptokinase. Data from the Hirulog and Early Reperfusion/Occlusion (HERO) 1 trial (n = 412) indicate that bivalirudin recipients were significantly more likely to have Thrombin Inhibition in Myocardial Ischaemia (TIMI) grade 3 flow at 90 to 120 minutes than heparin recipients. In addition, data from the HERO-2 trial (n = 17 073) show bivalirudin was significantly more effective than heparin in reducing adjudicated 96-hour reinfarction and 30-day investigator-reported death/reinfarction than heparin. Bivalirudin was as effective as heparin in reducing 30-day mortality. Data from a meta-analysis of four randomised trials among patients undergoing PTCA or treatment for acute coronary syndromes indicate that, at after 30 to 50 days of follow-up, bivalirudin was significantly more effective than heparin in reducing the incidence of nonfatal MI and the combined endpoint of death or nonfatal MI. The most significant adverse events associated with bivalirudin are bleeding complications. In individual trials, bivalirudin was as well tolerated as heparin with, in general, a reduced incidence of bleeding complications. Additionally, bivalirudin provides a more consistent, predictable anticoagulant response. In 4312 patients with unstable angina undergoing PTCA the incidence of retroperitoneal bleeding, blood transfusion and major haemorrhage was significantly lower in bivalirudin than heparin recipients. Data from the HERO-2 trial in patients with acute MI indicate that although bivalirudin recipients had a significantly higher incidence of mild or moderate bleeding than heparin recipients, there was no difference in intracranial haemorrhage, severe bleeding or transfusions. Data from a meta-analysis among 5674 patients with ischaemic heart disease show bivalirudin recipients were at a significantly lower risk of haemorrhagic events than heparin recipients. CONCLUSIONS: Bivalirudin is an effective alternative to heparin in the prevention of ischaemic complications in patients with unstable angina undergoing PTCA. In addition, the drug has shown potential in the treatment of patients with unstable angina not undergoing PCI. For patients with MI, it is clear that bivalirudin can replace heparin in the management of MI where streptokinase is used as the thrombolytic agent. Further data are required on the efficacy of bivalirudin in patients undergoing thrombolysis with newer thrombolytics. PMID- 11929335 TI - Treatment of sexually transmitted infections with single-dose therapy: a double edged sword. AB - Since the advent of the antimicrobial era, single-dose therapy has been a valuable tool in the management of genital infection. Most of the common sexually transmitted infections (STIs) such as gonorrhoea, syphilis, trichomoniasis and chancroid can be treated in this way, as can genital infections which are not sexually transmitted such as bacterial vaginosis and genital tract candidiasis. Until recently, treatment for Chlamydia trachomatis infection required a multi dose regimen, but single-dose azithromycin has now been shown to be an effective and acceptable alternative to this. Unfortunately, eradicative therapy has proven to be elusive for the viral STIs such as genital herpes simplex infection, human papilloma virus infection and human immunodeficiency virus (HIV) infection. The main advantage of single-dose therapy lies in its convenience and in its ability to ensure virtually 100% compliance. This addresses the problems of reduced clinical efficacy and the difficulties in assessing the response to therapy which complicates poor treatment compliance. However, some single-dose regimens for STIs do have drawbacks, particularly in certain situations. This may be with respect to efficacy, for example in syphilis with single-dose benzathine penicillin therapy, particularly for pregnant women and individuals infected with HI. Alternatively, it may involve toxicity, for example with single-dose metronidazole therapy for trichomoniasis or bacterial vaginosis where a higher rate of gastrointestinal adverse effects may be expected than if a lower multi dose regimen is used. In addition, single-dose therapy, for example with nevirapine, given to the mother in labour and to the baby after delivery significantly reduces the risk of mother to child HIV transmission, but resistance mutations are frequently detected in the viral genome after the brief exposure to the drug, which could jeopardise its future use. Single-dose therapy clearly has both advantages and disadvantages. We have reviewed a range of these in a variety of situations, focussing on their applications, effectiveness, compliance and toxicity, highlighting how single-dose therapy may be a double edged sword. PMID- 11929326 TI - Chemoprevention of breast cancer: implications for postmenopausal women. AB - Estrogen administration is associated with reduction in perimenopausal symptoms and the risk for several conditions affecting postmenopausal women. As estrogen administration also increases the risk for breast cancer, a common dilemma facing many women and their physicians is whether to use estrogen replacement therapy (ERT), a selective estrogen receptor modulator (SERM) that antagonises estrogenic effects in breast tissue but retains some estrogen agonist properties in other organs, or neither. For women with average to moderate risk of breast cancer and with perimenopausal symptoms, ERT may be the best short-term choice. For very high-risk women (>1% per year) with menopausal symptoms, alternatives to ERT might be offered and tried first. A diagnosis of ductal carcinoma in situ or invasive breast cancer within the last 2 to 5 years should be considered a relative contraindication for ERT unless the tumour was estrogen receptor negative. High-risk women without menopausal symptoms are the best candidates for the only currently approved drug for breast cancer risk reduction, tamoxifen. Although the drug is approved for women with a 5-year risk of breast cancer > or = 1.7% (0.34% per year), postmenopausal women most likely to experience a favourable benefit/risk ratio are those with a Gail estimated risk of >0.5% per year without a uterus or >1% per year if they retain their uterus. Tamoxifen should not be used in women with prior history of thromboembolic or precancerous uterine conditions. Tamoxifen is often used in Europe in conjunction with transdermal ERT in hysterectomised women without obvious loss of efficacy or increased risk of thromboembolism. Raloxifene is a second generation SERM with estrogen-like agonist effects on bone but with less uterine estrogen agonist activity than tamoxifen. Raloxifene may have less potent breast antiestrogenic effects than tamoxifen, particularly in a moderate- to high-estrogen environment. Raloxifene is approved for use in reducing risk of osteoporosis, but not breast cancer. Whether it is as effective as tamoxifen in reducing breast cancer risk in postmenopausal women is the subject of a current trial. All women regardless of breast cancer risk are advised to employ nonpharmacological risk reduction measures, including normalisation of bodyweight, exercise, adequate calcium and vitamin D intake, and avoidance of smoking and alcohol. The preventive options are best weighed during an individualised consultation where a woman's menopausal symptoms and risk for breast cancer and other diseases can be examined, and the options for improving postmenopausal health can be discussed. PMID- 11929336 TI - Novel pharmacotherapeutic approaches to prevention and treatment of GVHD. AB - Acute and chronic graft versus host disease (GVHD) remain the major barriers to successful hematopoietic cell transplantation. The induction of GVHD may be divided into three phases: recipient conditioning;donor T-cell activation; andeffector cells mediating GVHD. This review examines GVHD prevention and treatment using this conceptual model as framework. The various pharmacological agents discussed impact on different phases of the GVHD cascade. For example, keratinocyte growth factor and interleukin (IL)-11 are cytokines that may be useful in disrupting phase I of the GVHD cascade by blocking gastrointestinal tract damage, and lowering serum levels of lipopolysaccharide and tumour necrosis factor (TNF)-alpha. Cyclosporin, tacrolimus (FK-506) and sirolimus (rapamycin) are some of the main agents that disrupt phase II (donor T-cell activation). Mycophenolate mofetil and tresperimus probably act on this phase as well. Other novel drugs that affect phase II are tolerance-induction agents such as CTLA-4 and anti-CD40-ligand monoclonal antibodies, and preliminary results using CTLA-4 monoclonal antibody in GVHD prevention are encouraging. Examples of agents that disrupt phase III are the IL-2 receptor antagonist daclizumab and the anti TNFalpha monoclonal antibody infliximab. These anti-cytokine antibodies have shown promising results in early studies. The most effective approach to GVHD prevention will probably be a combination regimen where the three phases of the GVHD cascade are disrupted. Once GVHD has occurred, all three phases of the cascade are activated. Developments of combination therapy for treatment of both acute and chronic GVHD are likely to yield better results than monotherapy. The numerous new treatment modalities presented should improve the outlook for patients with acute and chronic GVHD. PMID- 11929337 TI - Newer immunosuppressive drugs: their potential role in rheumatoid arthritis therapy. AB - Rheumatoid arthritis (RA) is a chronic immune-mediated disease characterised by chronic synovitis, which leads to cartilage damage and joint destruction. It is generally a progressive disease with radiographic evidence of joint damage, functional status decline and premature mortality. Proinflammatory cytokines, such as interleukin 1 and tumour necrosis factor alpha, play an important role in maintaining the chronicity of RA and mediating tissue damage. New approaches in the therapy of RA with anticytokine biological agents, which neutralise or block cytokines or their receptors, are now the first generation antirheumatic drugs in clinical practice. A better understanding of the signal transduction systems and gene regulation by transcription factors involved in cytokine production has opened the way for the discovery of novel therapeutic compounds useful in treating patients with RA. Overactivation of selective kinases or aberrant function of downstream transcription factors could help convert a normal immune response to a chronic disease state. This provides a unique opportunity for novel therapeutic interventions, since specific signal transduction or transcription factor targets might interrupt the perpetuation mechanisms in RA. The availability of potent and selective p38 mitogen activated protein kinase inhibitors provide a means in further dissecting the pathways implicated in cytokine production, which in turn maintain the chronicity of RA. Many studies conclude that these compounds are very useful in the treatment of chronic synovitis and therefore are very promising for RA treatment. PMID- 11929338 TI - Infections in neurosurgery: using laboratory data to plan optimal treatment strategies. AB - Laboratory services contribute to the management of patients with neurosurgical infections in a variety of ways and, in so doing, increase the likelihood of a favourable outcome. Microbiology laboratories and clinical microbiologists are able to confirm the diagnosis, identify the causative agents and facilitate optimal antimicrobial therapy. Other pathology specialties perform investigations which help neurosurgeons to differentiate between postoperative aseptic and bacterial meningitis, these disease processes being indistinguishable on clinical grounds. A broad range of variables have been evaluated to date, but only the lactate and interleukin-1beta concentrations in cerebrospinal fluid have been shown to have sufficiently high sensitivities and specificities to be useful for this purpose. In preliminary studies measurement of the serum C-reactive protein concentration has been shown to be an effective criterion for monitoring the response to antibacterial therapy in patients with spinal extradural abscesses, postoperative discitis, brain abscesses and subdural empyemas, thereby enabling patients to be treated successfully with courses of these drugs that are markedly shorter than those currently recommended. PMID- 11929340 TI - NSAIDs and gastrointestinal cancer prevention. AB - Numerous studies report the relationship between aspirin and other nonsteroidal anti-inflammatories (NSAIDs) and cancer incidence, in particular for colorectal cancer. This paper systematically reviews the evidence of the effect of aspirin and other NSAIDs on the primary prevention of colorectal and other gastrointestinal cancers in the general population. In 25 investigations of NSAIDs and colorectal cancer, 23 observational studies reported a relative risk reduction but estimates vary widely. Cohort studies generally indicate lesser reductions than case-control studies suggesting possible biases in the latter. Clear evidence of a dose relationship generally appears lacking but data do not indicate useful effects of aspirin in cardioprophylactic doses. Differences have otherwise not been detected between aspirin and other NSAIDs, nor between non aspirin NSAIDs. There is some evidence that the risk of colorectal cancer reduces with increased duration of NSAID use. The lower incidence of oesophageal and gastric cancers results in smaller numbers of cases in the studies reporting these cancers, particularly in the cohort studies. The trend is for a risk reduction for oesophageal and gastric cancers in people taking NSAIDs, which is more likely to be statistically significant in the case-control studies. A very small number of observational studies have reported the relationship between NSAIDs and the incidence of pancreatic, gallbladder and liver cancers. These show no consistent relationship. In view of the inadequate information about optimal dose and duration of NSAIDs for colorectal cancer reduction, and the adverse effects of NSAIDs, we are not yet in a position to recommend NSAIDs for the primary prevention of colorectal cancer in the general population. PMID- 11929342 TI - Moxifloxacin in uncomplicated skin and skin structure infections. AB - Moxifloxacin is a fluoroquinolone antibacterial agent which attains good penetration into peripheral tissues and inflammatory fluids. The drug shows good in vitro activity against staphylococci and streptococci. Moxifloxacin is therefore a suitable option for the treatment of uncomplicated skin and skin structure infections of bacterial origin. In clinical trials, moxifloxacin was as effective as cephalexin in the treatment of uncomplicated skin and skin structure infections in patients aged >or=18 years. Moxifloxacin 400mg once daily or cephalexin 500mg three times daily for 7 days both resulted in clinical resolution in 84% of patients during a double-blind, randomised trial in 401 patients (intent-to-treat). The main infectious agent in this study was Staphylococcus aureus. Similar results were obtained in two other randomised, double-blind trials published as abstracts. The bioavailability of moxifloxacin is substantially reduced by coadministration with antacids or iron preparations. Moxifloxacin, however, does not show pharmacokinetic interaction with theophylline or warfarin. Dosage adjustments are not required in patients with renal impairment or in patients with mild to moderate hepatic insufficiency. The most common adverse events reported during moxifloxacin treatment are gastrointestinal disturbances. The potential for photosensitivity reactions during moxifloxacin treatment is low. PMID- 11929341 TI - Tamoxifen resistant and refractory breast cancer: the value of aromatase inhibitors. AB - Tamoxifen has dominated endocrine treatment of breast cancer for over two decades. It is useful in metastatic breast cancer, adjuvant therapy, preoperative treatment, ductal carcinoma-in-situ and chemoprevention. However, breast cancer may be refractory to tamoxifen or develop resistance to it with ongoing treatment. This resistance involves several mechanisms including receptor mutation causing 'estrogen hypersensitivity' and an increasing agonist effect of tamoxifen. Megestrol (megestrol acetate), in North America, and aminoglutethimide, in Europe, have been the traditional second line therapies after tamoxifen in advanced breast cancer. Aromatase (estrogen synthetase) inhibitors are a logical alternative to tamoxifen to antagonise the effects of estrogen on breast cancer. The third-generation non-steroidal aromatase inhibitors anastrozole, letrozole and vorozole, and the steroidal inhibitor exemestane, have been studied after tamoxifen versus either megestrol or aminoglutethimide. They showed enhanced efficacy and significantly superior toxicity profiles. Compliance with the inhibitors was also significantly better than with the traditional treatments. Aromatase inhibitors have most recently been shown to be superior to tamoxifen as initial therapy and are being extensively tested in the adjuvant setting after, or instead of, tamoxifen. Pilot studies of chemoprevention are also being undertaken. The aromatase inhibitors are an important new addition to the armamentarium of breast cancer therapy. PMID- 11929345 TI - Optimising drug utilisation in long term care. AB - Providing quality long term care for the elderly while containing costs is presenting major challenges for governments and policy makers. Although international variability exists with respect to the number of medications and other factors influencing suboptimal pharmacotherapy, suboptimal pharmacotherapy among elderly persons is common. This international problem requires a creative and multifaceted approach to improve and rationalise prescribing. We outline the non-regulatory efforts and regulatory means to approaching this problem. The recent introduction of a prospective payment system for long-term care in the US has underscored the importance of a regulatory approach to counter-balance the cost containment efforts which bundle the cost of medications into a prospectively set per diem rate. An examination of how US regulatory bodies are considering improving prescribing is provided. Considering the case of coronary heart disease, we provide data regarding the performance of a quality indicator aimed at stimulating quality prescribing for this medical condition. Although the use of regulatory approaches can improve prescribing, it is also recognised that a more holistic approach involving multidisciplinary teams and greater focus on the patient is the ultimate aspiration. This is particularly the case with the elderly in whom appropriate drug therapy can have a major impact on outcomes. A major cultural shift in the way society views and treats the elderly may be required in order to produce dramatic improvements in long term care for older people. PMID- 11929346 TI - Acute exacerbations of chronic bronchitis: a pharmacoeconomic review of antibacterial use. AB - Chronic bronchitis is a common problem affecting a large proportion of the adult population. People with chronic bronchitis are subject to recurrent attacks of bronchial inflammation called acute exacerbations of chronic bronchitis (AECBs). In patients with AECBs, symptoms may worsen due to a bacterial infection; the exacerbation is then known as an acute bacterial exacerbation of chronic bronchitis (ABECB). ABECBs are thought to be controllable through the use of antibacterial agents. In this paper we review current evidence on the cost of chronic bronchitis and AECBs, the cost effectiveness of antibacterials in the management of ABECB, and the factors that may affect the cost-effectiveness of antibacterials in the management of ABECB. We find that the number of economic evaluations conducted in this area is small. Of the few economic evaluations that have been conducted there has been only one prospective economic evaluation based on a clinical trial. The remainder are simple decision analysis-based modelling studies or retrospective database studies. Our principle findings are as follows: a key factor affecting the cost-effective use of antibacterials in the management of ABECB is the definitive diagnosis of the condition. Unfortunately, diagnosing a bacterial cause of an AECB is difficult, which presents problems in ensuring that antibacterials are not prescribed unnecessarily;current evidence suggests but does not prove that use of more effective but more costly first-line antibacterials may be relatively cost effective and may minimise overall expenditure by reducing the high costs associated with treatment failure;chronic bronchitis and AECB have a significant and negative physical and psychological effect on health-related quality of life. In conclusion, the small number of economic evaluations conducted in this area, coupled with the nature of the design of these studies, precludes a definitive statement recommending which specific antibacterial should be preferred on cost-effectiveness grounds for the management of ABECB. On the basis of our findings we suggest some topics for further research. PMID- 11929347 TI - Intermittent claudication: pharmacoeconomic and quality-of-life aspects of treatment. AB - The purpose of this article is to review the literature on the pharmacoeconomics of treatment for intermittent claudication and to discuss the importance of quality-of-life assessment for evaluating treatment strategies. Systemic risk reduction is the primary objective in the treatment of patients with intermittent claudication, as these patients have a high future risk of cardiovascular morbidity and mortality. Modification of cardiovascular risk factors accompanied by antiplatelet therapy is likely to improve overall survival, reduce myocardial infarction and stroke, and will, perhaps, also reduce the risk of ulcers and amputation at acceptable cost-effectiveness ratios. The second goal in the treatment of patients with intermittent claudication is to improve their walking capacity and community-based functional status. Supervised exercise training is the most effective noninvasive intervention to improve walking capacity, but may have elevated indirect costs. Among patients with disabling claudication who are candidates for invasive therapeutic procedures, angioplasty is cost effective in those with femoropopliteal stenosis or occlusion and in those with critical limb ischaemia and a stenosis. For all these therapeutic strategies there is a need to relate the costs to a relevant and comprehensive measure of effectiveness. Quality-of-life evaluation by using questionnaires exploring the specific problems encountered by patients with intermittent claudication in their daily life appear to be the most appropriate tool to evaluate the net result of a treatment. Cost-utility studies by combining pecuniary and quality-of-life evaluations provide information that is extremely useful to patients with intermittent claudication, regulatory authorities, the pharmaceutical industry and healthcare providers. PMID- 11929348 TI - Development of an economic model to assess the cost effectiveness of asthma management strategies. AB - BACKGROUND AND OBJECTIVE: Asthma is a chronic-episodic disease characterised by acute, symptomatic episodes of varying severity. We developed a Markov model that can be used to estimate the cost effectiveness of alternative asthma treatments. Because of the costs they incur, asthma exacerbations ('attacks') requiring intervention by a healthcare professional were a central consideration in the development of the model. METHODS: Treatment success was assessed as asthma control, a composite measure based on goals defined in world-wide asthma management guidelines and in terms of quality-adjusted life-years (QALYs). The data from which the transition probabilities were derived came from patients with asthma who received either salmeterol/fluticasone propionate combination (SFC) 50/100microg or fluticasone propionate (FP) 100microg, administered twice daily via an inhaler, in a 12-week, randomised, double-blind, clinical trial. Costs were estimated from resource profiles defined for each of the model states. A key aspect of the model was the use of probabilistic sensitivity analysis techniques to examine the uncertainty in the cost-effectiveness results. Distributions were fitted to transition probabilities and to cost input parameters and values were sampled at random from these distributions using a second order Monte Carlo simulation technique. This produced a distribution for incremental cost effectiveness that was employed to construct 95% uncertainty intervals and to construct cost effectiveness acceptability curves. RESULTS: In this analysis, the model was run over a 12-week period using transition probabilities derived from the trial data. The results showed that treatment with SFC resulted in a higher proportion of successfully controlled weeks per patient than treatment with FP (66 vs 47%), and higher mean weekly direct asthma management costs (pound sterling 15.77 vs pound sterling 11.83; 2000 values). The average incremental cost per successfully controlled week with SFC was pound sterling 20.83. Probabilistic sensitivity analysis showed that the 95% uncertainty intervals for the incremental cost-effectiveness ratio was - pound sterling 64.94 to pound sterling 112.66. In approximately 25% of cases, SFC was dominant (more effective and less costly), but in the remaining cases, it was both more effective and more costly. It was shown that if decision makers are willing to pay approximately pound sterling 45 for an additional successfully controlled week, SFC will be the more cost-effective strategy in this patient population for 80% of the time. CONCLUSIONS: This is one of the first decision-analytic models of asthma to incorporate probabilistic sensitivity analysis techniques to explore uncertainty. The model's flexible yet standardised framework permits the cost effectiveness of alternative asthma management strategies in different healthcare settings to be established. PMID- 11929349 TI - Economic evaluation of triflusal and aspirin in the treatment of acute myocardial infarction. AB - OBJECTIVE: To compare the costs to the Spanish healthcare system of 35 days' treatment with triflusal (600 mg/day) and aspirin (300 mg/day) in patients with confirmed acute myocardial infarction within 24 hours of onset of symptoms. DESIGN: A cost minimisation analysis based on the results of the Triflusal in Acute Myocardial Infarction study (TIM) was conducted. The hypothesis was that despite a higher acquisition cost of triflusal, savings would result because of differences in efficacy and safety outcome (non-fatal cerebrovascular event and haemorrhagic events). Diagnostic Related Groups were used as a proxy for determining hospital costs in Spain and the values were obtained from different sources and refer to year 2000 costs. Only direct medical costs were considered for the economic analysis. RESULTS: Although the acquisition cost of triflusal was more expensive than that of aspirin, the cost of prevented events - non-fatal ischaemic cerebrovascular events and cerebral haemorrhages - entirely compensated for the cost of triflusal. The overall cost of treating patients with triflusal, compared with aspirin, represented a net saving of 28.4% per patient treated. CONCLUSION: Our study showed that triflusal is cost saving compared with aspirin in the treatment of the acute phase of myocardial infarction. PMID- 11929350 TI - Economic value of thrombolysis with adjunctive abciximab in patients with subacute peripheral arterial occlusion. AB - BACKGROUND AND OBJECTIVE: Glycoprotein (GP) IIb/IIIa receptor inhibitors enhance thrombolysis in patients with acute coronary syndromes. This analysis evaluates the economic impact of abciximab, a GP IIb/IIIa inhibitor, as an adjunct to urokinase in peripheral artery occlusions of less than 6 weeks duration. STUDY DESIGN: A post-hoc economic analysis was performed using clinical data and inpatient resource utilisation derived from the prospective comparative phase II Platelet Receptor Antibodies in Order to Manage Peripheral Artery Thrombosis (PROMPT) pilot study. Study endpoints were amputation-free survival and survival without open surgery or major amputation after 90 days, and the rate of major complications at 30 days. PERSPECTIVE: Third-party payer and the societal perspective. PATIENTS AND METHODS: Seventy patients with lower extremity thrombi were randomised (2 : 5 ratio) to urokinase plus placebo or to urokinase plus abciximab. Economically relevant data were retrospectively derived from the clinical study database from a specific evaluation of patient records and from expert opinion. RESULTS: From the viewpoint of the society, average total per patient direct and indirect costs accruing over 3 months were more favourable for treatment with abciximab plus urokinase than for urokinase alone [9723 euros (EUR) vs EUR10 322; 2000 values], despite higher initial hospitalisation costs of the combination therapy. Abciximab plus urokinase was the dominant strategy at 3 months due to a clinically higher rate of survival without amputation or bypass surgery coupled with a lower average per-patient cost. From the perspective of the third-party payer, treatment with abciximab plus urokinase was economically also superior to urokinase alone (EUR8773 vs EUR9663). CONCLUSIONS: Based on the preliminary findings of the PROMPT trial, the use of abciximab as an adjunct to urokinase in patients with subacute peripheral artery occlusions may be the favourable strategy compared with urokinase alone, in terms of clinical and economic outcomes. Further trials are needed to confirm these clinical and economic findings. The preliminary clinical benefits experienced by patients treated with abciximab plus urokinase in the PROMPT trial translated into cost savings in terms of reduced direct medical costs at 3 months. These cost savings more than offset the cost of abciximab. The use of abciximab as an adjunct to urokinase in patients with subacute peripheral artery occlusions may be the favourable strategy compared with urokinase-alone in terms of clinical and economic outcomes, but further trials are needed to confirm the these clinical and economic findings. PMID- 11929351 TI - Tai Chi Chuan: an ancient wisdom on exercise and health promotion. AB - Tai Chi Chuan (TCC) is a Chinese conditioning exercise and is well known for its slow and graceful movements. Recent investigations have found that TCC is beneficial to cardiorespiratory function, strength, balance, flexibility, microcirculation and psychological profile. The long-term practice of TCC can attenuate the age decline in physical function, and consequently it is a suitable exercise for the middle-aged and elderly individuals. TCC can be prescribed as an alternative exercise programme for selected patients with cardiovascular, orthopaedic, or neurological diseases, and can reduce the risk of falls in elderly individuals. The exercise intensity of TCC depends on training style, posture and duration. Participants can choose to perform a complete set of TCC or selected movements according to their needs. In conclusion, TCC has potential benefits in health promotion, and is appropriate for implementation in the community. PMID- 11929352 TI - Physiological profiles of elite senior wrestlers. AB - To perform excellently in international competitions, wrestlers have to achieve an excellent level of physical fitness and physical condition during training. This article reviews the physiological profiles of elite wrestlers. In general, successful wrestlers showed higher dynamic and isokinetic strength than unsuccessful wrestlers. In particular, upper body strength and anaerobic power were significantly different between the two groups. Aerobic capacity is one of the most important physical factors to achieve good results in wrestling competitions. The maximal oxygen uptake of national and international wrestlers taking part in international competition has been shown to be about 53 to 56 ml/kg/min. Around the time of the Seoul Olympics, typical values for wrestlers were about 60 ml/kg/min, with values of >70 ml/kg/min in some cases (the latter being similar to values reported for some endurance runners). The flexibility of the wrestlers was found to be lower than that of weight lifters and gymnasts. However, the flexibility of top-level wrestlers was higher than that of lower level wrestlers. To fully understand the physiological profiles of successful wrestlers, further research regarding anaerobic and aerobic energy metabolism, cardiopulmonary responses, body fat content, and changes in muscle hypertrophy both during the wrestling season and off-season is warranted. PMID- 11929339 TI - Pharmacological approaches for the treatment of obesity. AB - The high incidence of obesity, its multifactorial nature, the complexity and lack of knowledge of the bodyweight control system, and the scarcity of adequate therapeutics have fuelled anti-obesity drug development during a considerable number of years. Irrespective of the efforts invested by researchers and companies, few products have reached a minimum level of effectiveness, and even fewer are available in medical practice. As a consequence of anti-obesity research, our knowledge of the bodyweight control system increased but, despite this, the pharmacological approaches to the treatment of obesity have not resulted yet in effective drugs. This review provides a panoramic of the multiple different approaches developed to obtain workable drugs. These approaches, however, rely in only four main lines of action: control of energy intake, mainly through modification of appetite;control of energy expenditure, essentially through the increase of thermogenesis;control of the availability of substrates to cells and tissues through hormonal and other metabolic factors controlling the fate of the available energy substrates; andcontrol of fat reserves through modulation of lipogenesis and lipolysis in white adipose tissue. A large proportion of current research is centred on neuropeptidic control of appetite, followed by the development of drugs controlling thermogenic mechanisms and analysis of the factors controlling adipocyte growth and fat storage. The adipocyte is also a fundamental source of metabolic signals, signals that can be intercepted, modulated and used to force the brain to adjust the mass of fat with the physiological means available. The large variety of different approaches used in the search for effective anti-obesity drugs show both the deep involvement of researchers on this field and the large amount of resources devoted to this problem by pharmaceutical companies. Future trends in anti-obesity drug research follow closely the approaches outlined; however, the increasing mass of information on the molecular basis of bodyweight control and obesity will in the end prevail in our search for effective and harmless anti-obesity drugs. PMID- 11929354 TI - Return-to-work interventions for low back pain: a descriptive review of contents and concepts of working mechanisms. AB - Low back pain is a major medical and social problem associated with disability, work absenteeism and high costs. Given the impact of the problem, there is a need for effective treatment interventions in occupational healthcare that aim at the prevention of chronic disability and the realisation of return to work. These so called return-to-work (RTW) interventions are becoming increasingly popular. As well as questions concerning the effectiveness of RTW interventions, there are also important questions on the actual content and underlying concepts of these multifactorial intervention strategies. The purpose of this review is to examine the literature on the content and underlying concepts of RTW interventions for low back pain. A systematic literature search identified 14 randomised controlled trials (RCTs) evaluating the effects of 19 RTW interventions. The content and concepts of these RTW interventions are described, compared and discussed in this review. Further, the contents of the RTW interventions are classified by the use of predefined components (physical exercises, education, behavioural treatments and ergonomic measures). The identified RTW interventions varied with respect to the disciplines involved, the target population and the number and duration of sessions. The classification showed that physical exercises were a component of most of the selected interventions, followed by education, behavioural treatments and ergonomic measures. The most prevalent combination of components was the combination of physical exercises, behavioural treatment and education. However, the types of physical exercises, behavioural treatment and education varied widely among the RTW interventions. The described concepts for the physical exercises were an increase of muscle strength, coordination, range of motion of the spine and cardiovascular fitness, and a decrease of muscle tension. Education as a part of RTW interventions is believed to increase the understanding of patients regarding their disorder and treatment. Behavioural treatments were mainly based on the gate control theory of pain (psychophysiological processes are involved in pain perception) and/or the operant conditioning hypothesis (pain behaviour is determined by its consequences). No concepts were described for ergonomic measures. Finally, the plausibility of the described concepts is discussed. Future RCTs on this topic should evaluate the underlying concepts of the RTW intervention in addition to its effectiveness. PMID- 11929353 TI - Musculoskeletal problems of the chest wall in athletes. AB - Chest pain in the athlete has a wide differential diagnosis. Pain may originate from structures within the thorax, such as the heart, lungs or oesophagus. However, musculoskeletal causes of chest pain must be considered. The aim of this review is to help the clinician to diagnose chest wall pain in athletes by identifying the possible causes, as reported in the literature. Musculoskeletal problems of the chest wall can occur in the ribs, sternum, articulations or myofascial structures. The cause is usually evident in the case of direct trauma. Additionally, athletes' bodies may be subjected to sudden large indirect forces or overuse, and stress fractures of the ribs caused by sporting activity have been extensively reported. These have been associated with golf, rowing and baseball pitching in particular. Stress fractures of the sternum reported in wrestlers cause pain and tenderness of the sternum, as expected. Diagnosis is by bone scan and limitation of activity usually allows healing to occur. The slipping rib syndrome causes intermittent costal margin pain related to posture or movement, and may be diagnosed by the 'hooking manoeuvre', which reproduces pain and sometimes a click. If reassurance and postural advice fail, good results are possible with resection of the mobile rib. The painful xiphoid syndrome is a rare condition that causes pain and tenderness of the xiphoid and is self limiting. Costochondritis is a self-limiting condition of unknown aetiology that typically presents with pain around the second to fifth costochondral joints. It can be differentiated from Tietze's syndrome in which there is swelling and pain of the articulation. Both conditions eventually settle spontaneously although a corticosteroid injection may be useful in particularly troublesome cases. The intercostal muscles may be injured causing tenderness between the ribs. Other conditions that should be considered include epidemic myalgia, precordial catch syndrome and referred pain from the thoracic spine. PMID- 11929355 TI - A conceptual framework for achieving performance enhancing drug compliance in sport. AB - There has been, and continues to be, widespread international concern about athletes' use of banned performance enhancing drugs (PEDs). This concern culminated in the formation of the World Anti-Doping Agency (WADA) in November 1999. To date, the main focus on controlling the use of PEDs has been on testing athletes and the development of tests to detect usage. Although athletes' beliefs and values are known to influence whether or not an athlete will use drugs, little is known about athletes' beliefs and attitudes, and the limited empirical literature shows little use of behavioural science frameworks to guide research methodology, results interpretation, and intervention implications. Mindful of this in preparing its anti-doping strategy for the 2000 Olympics, the Australian Sports Drug Agency (ASDA) in 1997 commissioned a study to assess the extent to which models of attitude-behaviour change in the public health/injury prevention literature had useful implications for compliance campaigns in the sport drug area. A preliminary compliance model was developed from three behavioural science frameworks: social cognition models; threat (or fear) appeals; and instrumental and normative approaches. A subsequent review of the performance enhancing drug literature confirmed that the overall framework was consistent with known empirical data, and therefore had at least face validity if not construct validity. The overall model showed six major inputs to an athlete's attitudes and intentions with respect to performance enhancing drug usage: personality factors, threat appraisal, benefit appraisal, reference group influences, personal morality and legitimacy. The model demonstrated that a comprehensive, fully integrated programme is necessary for maximal effect, and provides anti-doping agencies with a structured framework for strategic planning and implementing interventions. Programmes can be developed in each of the six major areas, with allocation of resources to each area based on needs-assessment research with athletes and other relevant groups. PMID- 11929357 TI - Blood lipids and lipoproteins in child and adolescent athletes. AB - The purpose of this review is to describe the age- and gender-associated variation in blood lipids of young athletes and examine the association between training status, peak oxygen consumption (VO(2peak)), and body fatness with blood lipids in young athletes. Results from cross-sectional studies suggest similar levels of total cholesterol (TC), lower levels of triglycerides (TG) and low density lipoprotein-cholesterol (LDL-C), and higher levels of high-density lipoprotein-cholesterol (HDL-C) in young athletes compared with controls. Longitudinal data show that the age- and gender-associated variation in blood lipids in young distance runners is similar to youth in the general population; TC and LDL-C remain stable, HDL-C declines during adolescence (especially in males), and TG increases with age. Considerable heterogeneity exists in the blood lipids of young athletes, including dyslipidaemic values. Age, sexual maturity status, training status, VO(2peak) and body fatness are determinants of blood lipids in young athletes. VO(2peak) is a significant predictor of HDL-C independent of body fatness. Further research is warranted to examine: the prevalence rates of dyslipidaemia in various sport groups; and the complex interactions of genes, biological maturation, exercise training, dietary intake and composition, skeletal muscle and adipose tissue properties on lipoprotein metabolism in young athletes. PMID- 11929356 TI - Anabolic steroids: a review for the clinician. AB - The number of athletes self-administering ergogenic pharmacological agents to increase their competitive edge continues to be a problem. Most athletes using anabolic steroids (AS) have acquired a crude pharmacological database regarding these drugs. Their opinions regarding steroids have been derived from their subjective experiences and anecdotal information. For this reason, traditional warnings regarding the lack of efficacy and potential dangers of steroid misuse are disregarded. A common widely held opinion among bodybuilders is that the anabolic steroid experts are the athletic gurus who for years have utilised themselves as the experimental participants and then dispensed their empirical findings. This review will address the common anabolic steroid misconceptions held by many of today's athletes by providing an evaluation of the scientific literature related to AS in athletic performance. PMID- 11929358 TI - Dermatologic disorders of the athlete. AB - The most common injuries afflicting the athlete affect the skin. The list of sports-related dermatoses is vast and includes infections, inflammatory conditions, traumatic entities, environmental encounters, and neoplasms. It is critical that the sports physician recognises common and uncommon skin disorders of the athlete. Knowledge of the treatment and prevention of various sports related dermatoses results in prompt and appropriate care of the athlete. Infections probably cause the most disruption to individual and team activities. Herpes gladiatorum, tinea corporis gladiatorum, impetigo, and furunculosis are sometimes found in epidemic proportions in athletes. Vigilant surveillance and early treatment help teams avoid these epidemics. Fortunately, several recent studies suggest that pharmacotherapeutic prevention may be effective for some of these sports-related infections. Inflammatory cutaneous conditions may be banal or potentially life threatening as in the case of exercise-induced anaphylaxis. Athletes who develop exercise-induced anaphylaxis may prevent outbreaks by avoiding food before exercise and extreme temperatures while they exercise. Almost all sports enthusiasts are at risk of developing traumatic entities such as nail dystrophies, calluses and blisters. Other more unusual traumatic skin conditions, such as talon noire, jogger's nipples and mogul's palm, occur in specific sports. Several techniques and special clothing exist to help prevent traumatic skin conditions in athletes. Almost all athletes, to some degree, interact with the environment. Winter sport athletes may develop frostbite and swimmers in both fresh and saltwater may develop swimmer's itch or seabather's eruption, respectively. Swimmers with fair skin and light hair may also present with unusual green hair that results from the deposition of copper within the hair. Finally, athletes are at risk of developing both benign and malignant neoplasms. Hockey players, surfers, boxers and football players can develop athlete's nodules. Outdoor sports enthusiasts are at greater risk of developing melanoma and non-melanoma skin cancer. Athletes spend a great deal of time outdoors, typically during peak hours of ultraviolet exposure. The frequent use of sunscreens and protective clothing will decrease the athlete's sun exposure. It is critical that the sports physician recognises common and uncommon skin disorders of the athlete. Knowledge of the treatment and prevention of various sports-related dermatoses results in prompt and appropriate care of the athlete. PMID- 11929359 TI - Effect of dietary intake on immune function in athletes. AB - Athletes are exposed to acute and chronic stress that may lead to suppression of the immune system and increased oxidative species generation. In addition, the tendency to consume fewer calories than expended and to avoid fats may further compromise the immune system and antioxidant mechanisms. The exercise stress is proportional to the intensity and duration of the exercise, relative to the maximal capacity of the athlete. Muscle glycogen depletion compromises exercise performance and it also increases the stress. Glycogen stores can be protected by increased fat oxidation (glycogen sparing). The diets of athletes should be balanced so that total caloric intake equals expenditure, and so that the carbohydrates and fats utilised in exercise are replenished. Many athletes do not meet these criteria and have compromised glycogen or fat stores, have deficits in essential fats, and do not take in sufficient micronutrients to support exercise performance, immune competence and antioxidant defence. Either overtraining or under nutrition may lead to an increased risk of infections. Exercise stress leads to a proportional increase in stress hormone levels and concomitant changes in several aspects of immunity, including the following: high cortisol; neutrophilia; lymphopenia; decreases in granulocyte oxidative burst, nasal mucociliary clearance, natural killer cell activity, lymphocyte proliferation, the delayed-type sensitivity response, the production of cytokines in response to mitogens, and nasal and salivary immunoglobulin A levels; blunted major histocompatibility complex II expression in macrophages; and increases in blood granulocyte and monocyte phagocytosis, and pro- and anti-inflammatory cytokines. In addition to providing fuel for exercise, glycolysis, glutaminlysis, fat oxidation and protein degradation participate in metabolism and synthesis of the immune components. Compromising, or overusing, any of these components may lead to immunosuppression. In some cases, supplementation with micronutrients may facilitate the immune system and compensate for deficits in essential nutrients. In summary, athletes should eat adequate calories and nutrients to balance expenditure of all nutrients. Dietary insufficiencies should be compensated for by supplementation with nutrients, with care not to over compensate. By following these rules, and regulating training to avoid overtraining, the immune system can be maintained to minimise the risk of upper respiratory tract infections. PMID- 11929360 TI - Adductor muscle strains in sport. AB - An in-season adductor muscle strain may be debilitating for the athlete. Furthermore, an adductor strain that is treated improperly could become chronic and career threatening. Any one of the six muscles of the adductor group could be involved. The degree of injury can range from a minor strain (Grade I), where minimal playing time is lost, to a severe strain (Grade III) in which there is complete loss of muscle function. Ice hockey and soccer players seem particularly susceptible to adductor muscle strains. In professional ice hockey players throughout the world, approximately 10% of all injuries are groin strains. These injuries, which have been linked to hip muscle weakness, previous injuries to that area, preseason practice sessions and level of experience, may be preventable if such risk factors can be addressed before each season. Hip strengthening exercises were shown to be an effective method of reducing the incidence of adductor strains in one closely followed National Hockey League ice hockey team. Despite the identification of risk factors and strengthening intervention for ice hockey players, adductor strains continue to occur throughout sport. Clinicians feel an active training programme, along with completely restoring the strength of the adductor muscle group, is the key to successful rehabilitation. Surgical intervention is available if nonoperative treatment fails for 6 months or longer. Adductor release and tenotomy was reported to have limited success in athletes. PMID- 11929361 TI - Intimate partner violence following pregnancy. PMID- 11929362 TI - Genetic epidemiology and congenital malformations: from the chromosome to the crib. PMID- 11929363 TI - Concurrent serious bacterial infections in 2396 infants and children hospitalized with respiratory syncytial virus lower respiratory tract infections. AB - BACKGROUND: At Driscoll Children's Hospital (Corpus Christi, Tex), we observed that most infants and children hospitalized for treatment of respiratory syncytial virus (RSV) bronchiolitis and/or pneumonia received broad-spectrum intravenous antibiotics despite having typical RSV signs and symptoms and positive RSV-rapid-antigen tests on admission. Physicians were concerned about the possibility of concurrent serious bacterial infections, especially in infants younger than 3 months and in those with infiltrates present on the chest x-ray films. OBJECTIVE: To report the frequency of concurrent serious bacterial infections in infants and children hospitalized for treatment of RSV lower respiratory tract infections. METHODS: The medical records of 2396 infants and children admitted to Driscoll Children's Hospital with RSV bronchiolitis and/or pneumonia during 7 RSV seasons from July 1, 1991, through June 30, 1998, were reviewed. RESULTS: There were positive cultures obtained from initial sepsis/meningitis workups on admission in 39 infants and children (1.6%). Of these, 12 (31%) were positive blood cultures and 27 (69%) were positive urine cultures. There were no positive cerebrospinal fluid cultures. All of the positive blood cultures contained either Staphylococcus epidermidis, Staphylococcus warneri, or Bacillus species, which are common contaminants. None of the patients received a standard 10-day course of intravenous antibiotic therapy. All of the positive urine cultures were typical urinary tract pathogens. All of the patients were treated. CONCLUSIONS: Concurrent serious bacterial infections are rare in infants and children hospitalized with RSV lower respiratory tract infections and the empiric use of broad-spectrum intravenous antibiotics is unnecessary in children with typical signs and symptoms of RSV bronchiolitis. PMID- 11929364 TI - Prevalence and patterns of intimate partner violence among adolescent mothers during the postpartum period. AB - OBJECTIVE: To examine prevalence, frequency, severity, and patterns of intimate partner violence (IPV) during the first 24 months' post partum within a multiethnic cohort of adolescents. DESIGN: A prospective study of adolescent girls followed up for 24 months into the postpartum period. Follow-up surveys were completed at 3, 6, 12, 18, and 24 months' post partum. Overall, 74% completed at least 4 of the 5 follow-up surveys. SETTING: Postpartum unit at a university teaching hospital in Galveston, Tex. PARTICIPANTS: A total of 570 adolescents (18 years or younger; 219 Mexican Americans, 182 African Americans, and 169 European Americans) completed face-to-face interviews within 48 hours of delivery and returned at least 4 of 5 follow-up surveys. MAIN OUTCOME MEASURES: Prevalence of IPV and frequent and severe IPV. RESULTS: Prevalence of IPV was highest at 3 months' post partum (21%) and lowest at 24 months (13%). The percentage of assaulted mothers who experienced severe IPV increased from 40% to 62% across this period. Seventy-five percent of mothers reporting IPV during pregnancy also reported IPV within 24 months following delivery. Of importance, 78% who experienced IPV during the first 3 postpartum months had not reported IPV before delivery. Ethnic differences in IPV were observed at 3, 6, and 18 months' post partum. CONCLUSIONS: Adolescents are at high risk for experiencing IPV during the postpartum period. Frequent screening for IPV by health care practitioners is critical to maximize detection. PMID- 11929365 TI - A statewide survey of domestic violence screening behaviors among pediatricians and family physicians. AB - OBJECTIVE: To assess rates of previous domestic violence (DV) training, current screening practices, and barriers to screening among Connecticut pediatric primary care physicians. DESIGN: Self-administered mail survey. SETTING: State of Connecticut. PARTICIPANTS: Pediatricians and pediatric care-providing family practice physicians (N = 903). RESULTS: The response rate was 49% (n = 438). The demographic characteristics of the response sample were as follows: 70% male, 76% older than 40 years, 84% white, 87% in private practice, and 64% in suburban practice. Only 12% of the physicians reported routinely screening for DV at all well-child care visits, 61% reported screening only selective patients, and 30% said they did not screen for DV at all. Sixteen percent of the physicians reported having an office protocol for dealing with victims of DV. Respondents practicing in an urban setting were significantly more likely to screen routinely for DV than those practicing in a suburban setting (odds ratio, 1.77; 95% confidence interval, 1.12-2.79). Prior DV training was the strongest predictor of routine screening (odds ratio, 5.17; 95% confidence interval, 3.13-8.56). In fact, respondents with previous training made up 64% of those who routinely screened for DV. CONCLUSIONS: Only a minority of Connecticut pediatric care physicians routinely screen mothers for DV. Primary care physicians with education and training about DV are screening at higher rates than physicians with no education and training. Pediatric physicians need training, protocols, and best-practice models on how to identify and intervene with families experiencing DV. PMID- 11929366 TI - Does soccer ball heading cause retinal bleeding? AB - OBJECTIVES: To define forces of youth soccer ball heading (headers) and determine whether heading causes retinal hemorrhage. SETTING: Regional Children's Hospital, youth soccer camp. PATIENTS: Male and female soccer players, 13 to 16 years old, who regularly head soccer balls. MEASUREMENTS: Dilated retinal examination, after 2-week header diary, and accelerometer measurement of heading a lofted soccer ball. RESULTS: Twenty-one youth soccer players, averaging 79 headers in the prior 2 weeks, and 3 players who did not submit header diaries lacked retinal hemorrhage. Thirty control subjects also lacked retinal hemorrhage. Seven subjects heading the ball experienced linear cranial accelerations of 3.7 +/- 1.3g. Rotational accelerations were negligible. CONCLUSIONS: Headers, not associated with globe impact, are unlikely to cause retinal hemorrhage. Correctly executed headers did not cause significant rotational acceleration of the head, but incorrectly executed headers might. PMID- 11929367 TI - Opportunities for appropriate care: health care and contraceptive use among adolescents reporting unwanted sexual intercourse. AB - BACKGROUND: Unwanted sexual contact, reported by 30% to 42% of young women and 10% to 34% of young men, has been associated with negative health outcomes and increased teenaged pregnancy. OBJECTIVE: To determine health services and contraceptive use among adolescents reporting unwanted sexual intercourse. METHODS: Random-digit dial methods were used to survey 1040 adolescents in Monroe County, New York; 389 (37%) were sexually active and answered a question about whether they had ever been forced or pressured to have sexual intercourse. The data were weighted to reflect the county population. RESULTS: Among sexually active adolescents, 20% of females and 7% of males reported unwanted intercourse (P<.001). For 37% of male and 17% of female adolescents, the survey was the first time they had disclosed the incident (P =.17). Among female adolescents reporting unwanted intercourse, 91% have a usual source of care and 62% reported a well visit in the previous 6 months. Female adolescents reporting unwanted sex were more likely to have wanted contraceptives but not gotten them because of fear their parents would find out (32% vs 11%; P =.01) and to have had sex without contraception (69% vs 52%; P =.05) than those who had not had unwanted sex. CONCLUSIONS: Many adolescents have been forced or pressured to have sexual intercourse. Although many have never told anyone about the incident, most have visited a primary care physician or clinician. Physicians and other clinicians should screen for a history of unwanted intercourse and provide needed referrals for counseling and/or contraceptive information. PMID- 11929368 TI - Children and adolescents with acute alcohol intoxication/self-poisoning presenting to the emergency department. AB - OBJECTIVE: To describe the presentations, characteristics, and follow-up care of children and adolescents aged 10 to 18 years who present to emergency departments (EDs) with acute alcohol intoxication/self-poisoning. DESIGN: Retrospective medical record review. SETTING: Five EDs in Western Sydney, Australia. PARTICIPANTS: Patients aged 10 to 18 years who presented to EDs with acute alcohol intoxication/self-poisoning between January 1, 1996, and December 31, 2000. MAIN OUTCOME MEASURES: Frequency of presentations; presentation characteristics; psychosocial characteristics; and presence or absence of follow up. RESULTS: Two hundred twelve children and adolescents presented to EDs 216 times. Of the 212 patients, 49 (23%) were 14 years or younger, and the youngest was aged 10 years. The majority (82%) came after hours and were brought in by emergency services (77%). In 13% of presentations, verbal and/or physical aggression was present, and a threat of self-harm was present in 2% of cases. A mental health worker was consulted about the child or adolescent in only 6% of presentations. Most children and adolescents (85%) were discharged from the ED. Of concern, in 56% of presentations, a follow-up plan was not recorded. There was documentation of mental health follow-up in only 14% of presentations and follow up from drug and alcohol services in only 1%. Forensic history, school functioning, and a history of past mental health problems were not documented in more than 60% of the medical records examined. CONCLUSIONS: When children or adolescents present to an ED with acute alcohol intoxication/self-poisoning, their risk factors for psychosocial dysfunction appear to be inadequately assessed, documented, and followed up. Clear guidelines for assessment and referral pathways must be established in EDs. PMID- 11929369 TI - Associations between health risk behaviors and opposite-, same-, and both-sex sexual partners in representative samples of vermont and massachusetts high school students. AB - OBJECTIVE: To examine associations between health risk behaviors and sexual experience with opposite-, same-, or both-sex partners in representative samples of high school students. DESIGN: We used 1995 and 1997 data from the Vermont and Massachusetts Youth Risk Behavior Surveys. Logistic regression and multiple regression analyses were used to compare health risk behaviors among students who reported sex with opposite-sex partners only (opposite-sex students), with same sex partners only (same-sex students), and with both male and female sexual partners (both-sex students). SETTING: Public high schools in Vermont and Massachusetts. PARTICIPANTS: Representative, population-based samples of high school students. The combined samples had 14 623 Vermont students and 8141 Massachusetts students. MAIN OUTCOME MEASURE: Violence, harassment, suicidal behavior, alcohol and other drug use, and unhealthy weight control practices. RESULTS: In both states, both-sex students were significantly more likely to report health risk behaviors than were opposite-sex students. For example, both sex students had odds 3 to 6 times greater than opposite-sex students of being threatened or injured with a weapon at school, making a suicide attempt requiring medical attention, using cocaine, or vomiting or using laxatives to control their weight. In both states, same-sex students were as likely as opposite-sex students to report most health risk behaviors. CONCLUSION: Relative to opposite- and same sex students, both-sex students may be at elevated risk of injury, disease, and death by experiencing serious harassment and engaging in violence, suicidal behavior, alcohol and other drug use, and unhealthy weight control practices. PMID- 11929370 TI - Prospective identification and treatment of children with pediatric autoimmune neuropsychiatric disorder associated with group A streptococcal infection (PANDAS). AB - BACKGROUND: The current diagnostic criteria for pediatric autoimmune neuropsychiatric disorder associated with group A streptococcal infection (PANDAS) are pediatric onset, neuropsychiatric disorder (obsessive-compulsive disorder [OCD]) and/or tic disorder; abrupt onset and/or episodic course of symptoms; association with group A beta-hemolytic streptococcal (GABHS) infection; and association with neurological abnormalities (motoric hyperactivity or adventitious movements, including choreiform movements or tics). OBJECTIVE: To assess new-onset PANDAS cases in relation to acute GABHS tonsillopharyngitis. DESIGN: Prospective PANDAS case identification and follow-up. RESULTS: Over a 3 year period (1998-2000), we identified 12 school-aged children with new-onset PANDAS. Each patient had the abrupt appearance of severe OCD behaviors, accompanied by mild symptoms and signs of acute GABHS tonsillopharyngitis. Throat swabs tested positive for GABHS by rapid antigen detection and/or were culture positive. The GABHS serologic tests, when performed (n = 3), showed very high antideoxyribonuclease antibody titers. Mean age at presentation was 7 years (age range, 5-11 years). In children treated with antibiotics effective in eradicating GABHS infection at the sentinel episode, OCD symptoms promptly disappeared. Follow-up throat cultures negative for GABHS were obtained prospectively after the first PANDAS episode. Recurrence of OCD symptoms was seen in 6 patients; each recurrence was associated with evidence of acute GABHS infection and responded to antibiotic therapy, supporting the premise that these patients were not GABHS carriers. The OCD behaviors exhibited included hand washing and preoccupation with germs, but daytime urinary urgency and frequency without dysuria, fever, or incontinence were the most notable symptoms in our series (58% of patients). Symptoms disappeared at night, and urinalysis and urine cultures were negative. CONCLUSION: To our knowledge, this is the first prospective study to confirm that PANDAS is associated with acute GABHS tonsillopharyngitis and responds to appropriate antibiotic therapy at the sentinel episode. PMID- 11929371 TI - A comparison of health and fitness-related variables in a small sample of children of Japanese descent on 2 continents. AB - OBJECTIVE: To compare physical characteristics, health and fitness-related variables, and nutrient intake between children of Japanese ancestry living in the United States and Japan. DESIGN: Cross-sectional study. SETTING: Miami, Fla, and Tsukuba, Japan. SUBJECTS: Fourteen children of Japanese descent living in the United States and 14 sex- and age-matched children living in Japan. MAIN OUTCOME MEASURES: US and Japanese resident groups were compared on physical characteristics, health and fitness-related variables, and nutrient intake using the t test for paired samples. To assess differences between groups in variables not statistically significant, effect sizes were calculated using the Cohen d test of standardized differences. RESULTS: The following significant differences were found between US and Japanese resident groups, respectively: body mass index, 19.3 and 16.9, P =.02; percentage of body fat, 22.0% and 14.3%, P =.002; diastolic blood pressure, 65.8 and 58.9 mm Hg, P =.01; total cholesterol, 169.8 and 138.7 mg/dL (4.39 and 3.59 mmol/L, P =.001); low-density lipoprotein cholesterol, 108.2 and 88.0 mg/dL (2.80 and 2.28 mmol/L, P =.01); triglycerides, 92.5 and 59.0 mg/dL (1.04 and 0.67 mmol/L, P =.02); percentage of fat intake, 26.1% and 20.3%, P =.001; percentage of saturated fat intake, 7.9% and 6.1%, P<.002; percentage of carbohydrate intake, 57.9% and 63.9% (P =.004); vertical jump, 28.9 and 34.4 cm, P =.02; and flexibility, 58.2 and 42.6 cm, P =.002. Using the Cohen d test, US residents showed a moderately greater systolic blood pressure (107.5 vs 101.9 mm Hg, P =.10) and leg strength (81.5 vs 55.8 kg, P =.11) than did Japanese residents. CONCLUSIONS: A small sample (n = 14) of children of Japanese descent living in Florida showed more adverse health-related characteristics than did a comparable group of their peers living in Japan. The results are probably related to differences in their diets. It remains to be seen whether the differences in diets are related to where the children live. PMID- 11929372 TI - Effect of managed care on children's relationships with their primary care physicians: differences by race. AB - CONTEXT: Racial and ethnic disparities in the quality of the patient-provider relationship have been documented previously, but only among adults. Few studies have examined this aspect of primary care quality for children, and none has examined the role of managed care in mediating disparities. OBJECTIVE: To explore variations in the associations among 3 managed care policies and the sustained patient-provider relationship for children by racial and ethnic group. DESIGN, SETTING, AND PARTICIPANTS: Telephone survey of parents of a random sample of 413 children attending elementary school in a large school district spanning 3 cities in southern California. Self-reported managed care information was validated through contact with health plan representatives. MAIN OUTCOME MEASURE: Parent reporting of the strength of affiliation and interpersonal relationships among the child, the family, and the health care provider. RESULTS: Parents of Hispanic and black children reported significantly lower strength of affiliation than whites did (mean, 3.19 and 3.27 vs 3.57; P<.05 and P<.01, respectively). Asians and Hispanics reported significantly poorer interpersonal relationships with providers than whites did (mean, 3.35 and 3.38 vs 3.53; P<.05 for both). Managed care policies requiring patients to (1) sign up with a particular provider, (2) seek a referral for specialty care, and (3) stay within a network were associated with reductions in the interpersonal relationship. All minority groups, but not whites, reported significantly poorer relationships when managed care policies were present. Asian children were most sensitive to the managed care policies. CONCLUSIONS: Our data suggest that racial and ethnic minority children experience poorer patient-provider relationships compared with white children, even controlling for socioeconomic status and health system factors. Minority children are particularly sensitive to managed care policies that restrict patient freedom in choosing where to seek care. Implementing less restrictive policies may enhance the development of the patient-provider relationship. PMID- 11929373 TI - Demonstrated use of metered-dose inhalers and peak flow meters by children and adolescents with acute asthma exacerbations. AB - OBJECTIVES: To determine the ability of children and adolescents with acute asthma exacerbations to adhere to national guidelines for proper metered-dose inhaler (MDI) and peak flow meter (PFM) technique and to define characteristics associated with improper use. DESIGN: A prospective study in which the patients were instructed to use a placebo MDI or a PFM in the emergency department exactly as at home. Technique was graded on the basis of performance of specific steps recommended by national guidelines. SETTING AND PARTICIPANTS: Children and adolescents (aged 2-18 years) with acute asthma exacerbations in the emergency department of an urban children's hospital with acute asthma. RESULTS: Thirty three (45.2%) of 73 patients using an MDI (MDI group) demonstrated multiple steps improperly compared with 60 (44.4%) of 135 using an MDI with a holding chamber (MDI-HC group; P =.92). In the MDI group, young ages of the patients (P<.008) and the parents (P<.003) were associated with improper use. In the MDI-HC group, factors independently and significantly associated with improper use were no hospitalizations within the past year, parent assistance of the patient with MDI HC use, and nondaily use of the MDI-HC. Also, 165 (82.9%) of 199 children who, per national guidelines, should be using a PFM at home, did not. Eighty-two (73.9%) of 111 patients demonstrated perfect performance of all PFM steps. CONCLUSIONS: Among children with acute asthma, we found high rates of improper MDI use and PFM underuse. A greater emphasis must be placed on teaching methods to optimize drug delivery and to instruct patients about the importance of self monitoring of disease severity. PMID- 11929374 TI - Measuring quality of life in children with attention-deficit/hyperactivity disorder and their families: development and evaluation of a new tool. AB - OBJECTIVE: To psychometrically evaluate a new parent-completed questionnaire that measures the effect of attention-deficit/hyperactivity disorder (ADHD) on the everyday well-being of children and their families. SETTING: Using a mail out/mail-back method, the sample was drawn from the registry of an outpatient developmental and behavioral program of a large tertiary pediatric hospital. All children received medication for ADHD. PARTICIPANTS: Responses were received for 81 children of whom 60 (74%) were boys. An even split of questionnaires was returned for children with ADHD primarily inattentive (50%) and ADHD combined (50%). The condition of 70 patients (86%) had been diagnosed for 1 year or longer; 69 patients (89%) reported receiving medication. MAIN OUTCOME MEASURE: The ADHD Impact Module, HealthAct, Boston, Mass, developed with input from families, measures the effect of the disorder on the child's emotional-social well-being (Child Scale, 8 items) and the family (Home Scale, 10 items). RESULTS: The scales exceeded standard criteria for item convergent and discriminant validity. No floor effects and minimal (2%) ceiling effects were observed. Cronbach alpha was 0.88 and 0.93 (Child and Home Scales), respectively. Raw scale scores are transformed on a 0 through 100 continuum; a higher score indicates more favorable findings. Statistically significant differences (P<.000) were observed for ADHD inattentive vs ADHD combined on both scales (Child, 65.26 vs 48.86; Home, 72.79 vs 51.26). Better "success at home" scores were reported by parents of ADHD inattentive children (Child Scale, 62.12 vs 47.36, P =.00; Home Scale, 70.58 vs 47.01, P =.000). CONCLUSIONS: The ADHD Impact Module meets stringent psychometric standards. Further validation is required, but current evidence suggests it is a promising new questionnaire. PMID- 11929375 TI - Trends and toxic effects from pediatric clonidine exposures. AB - OBJECTIVE: To analyze the trends, demographics, and toxic effects associated with pediatric clonidine hydrochloride exposures reported to poison centers. DESIGN: Retrospective. SETTING AND PATIENTS: Clonidine-only exposures followed up to known outcome in children younger than 19 years reported to the American Association of Poison Control Center's database from January 1, 1993, through December 31, 1999. MAIN OUTCOME MEASURES: Frequency of exposures over time, acuity, reason, symptoms, management site, treatment, and outcome. RESULTS: There were 10 060 reported exposures with 57% reported for children younger than 6 years, 34% for children between 6 and 12 years old, and 9% for adolescents between 13 and 18 years old. In 1999 there were 2.5 times as many exposures as in 1993. In 6- through 12-year-olds, clonidine was the child's medication in 35% of the exposures, compared with 10% in children younger than 6 years and 26% in adolescents. The proportion of cases involving the child's medication increased over 7 years. While unintentional overdose was most common in children younger than 6 years, therapeutic errors and suicide attempts predominated in 6- through 12-year-olds and adolescents, respectively. In 6042 symptomatic children (60%), the most common symptoms were lethargy (80%), bradycardia (17%), hypotension (15%), and respiratory depression (5%). Most exposures resulted in no effect (40%) or minor effects (39%). Moderate effects occurred in 1907 children (19%), major effects in 230 children (2%); there was 1 fatality in a 23-month-old. CONCLUSIONS: While most of the clonidine exposures resulted in minimal toxic effects, serious toxic effects and death can occur. The trend toward increasing the number of exposures in children, especially with evidence of toxic effects in children receiving clonidine therapeutically, is cause for concern. PMID- 11929376 TI - Measuring the loss of autonomy over nicotine use in adolescents: the DANDY (Development and Assessment of Nicotine Dependence in Youths) study. AB - CONTEXT: There is no validated, theory-based tool for assessing the onset of nicotine dependence. However, the use of all addictive substances can result in a loss of autonomy. We propose that nicotine dependence begins when autonomy is lost, ie, when the sequelae of tobacco use, either physical or psychological, present a barrier to quitting. OBJECTIVES: To test the autonomy theory of nicotine dependence, and to evaluate the Hooked on Nicotine Checklist (HONC) as a measure of the loss of autonomy over tobacco use. DESIGN: The psychometric performance and concept validity of the HONC were evaluated in a 30-month prospective longitudinal study of the natural history of tobacco use in a cohort of 679 seventh-grade students. RESULTS: As hypothesized, endorsement of a single item on the HONC was associated with a failed attempt at smoking cessation (odds ratio [OR], 29; 95% confidence interval [CI], 13-65), continued smoking until the end of follow-up (OR, 44; 95% CI, 17-114), and daily smoking (OR, 58; 95% CI, 24 142). Scores on the HONC correlated with the maximum amount smoked (r = 0.65; P<.001) and the maximum frequency of smoking (r = 0.79; P<.001). Internal reliability was 0.94. A 1-factor solution explained 66% of the total variance. CONCLUSIONS: The data support the autonomy theory that dependence begins with the loss of autonomy. The autonomy theory represents a potentially useful alternative to current concepts of nicotine dependence for adolescents, and the HONC appears to measure lost autonomy in adolescents. Construct validity was demonstrated by its utility in predicting failed cessation and the progression of tobacco use. In addition, the psychometric properties were excellent. PMID- 11929377 TI - Pathological case of the month. Type IV sacrococcygeal teratoma with yolk sac (endodermal sinus) tumor presenting with failure to thrive and perianal pain. PMID- 11929378 TI - Radiological case of the month. Occult spinal dysraphism (tethered cord, diastematomyelia, and spinal lipoma). PMID- 11929379 TI - Concealed weapons and children's hospitals: a new look at safety in our changing world. PMID- 11929382 TI - Review article: monitoring for drug side-effects in inflammatory bowel disease. AB - The side-effects suitable for monitoring in patients with inflammatory bowel disease being treated with the four main groups of drugs (5-aminosalicylic acid preparations, azathioprine and 6-mercaptopurine, methotrexate, and corticosteroids) are reviewed. On the basis of the reported frequency, severity and timing of side-effects, a practical scheme of monitoring is recommended. This includes a baseline measurement of full blood count, creatinine and liver function tests in all patients. In the absence of worrying symptoms, we recommend the following: (i) no monitoring for sulfasalazine; (ii) for other 5 aminosalicylic acid preparations, the measurement of creatinine at 6 and 12 months and then annually; (iii) for azathioprine/6-mercaptopurine, thiopurine methyltransferase genotype/phenotype determination has no role in treatment monitoring, but a full blood count at 2 weeks, 1 month, 3 months and then every 3 months should be performed; (iv) for methotrexate, a full blood count and liver function tests should be performed every 3 months; (v) for steroids, dual energy X-ray absorptiometry bone scanning should be performed at the start of therapy, every year in which steroids are used if the T score is < 0, and every 3-5 years if the T score is > 0. PMID- 11929383 TI - Review article: the concept of entero-colonic encephalopathy, autism and opioid receptor ligands. AB - There is growing awareness that primary gastrointestinal pathology may play an important role in the inception and clinical expression of some childhood developmental disorders, including autism. In addition to frequent gastrointestinal symptoms, children with autism often manifest complex biochemical and immunological abnormalities. The gut-brain axis is central to certain encephalopathies of extra-cranial origin, hepatic encephalopathy being the best characterized. Commonalities in the clinical characteristics of hepatic encephalopathy and a form of autism associated with developmental regression in an apparently previously normal child, accompanied by immune-mediated gastrointestinal pathology, have led to the proposal that there may be analogous mechanisms of toxic encephalopathy in patients with liver failure and some children with autism. Aberrations in opioid biochemistry are common to these two conditions, and there is evidence that opioid peptides may mediate certain aspects of the respective syndromes. The generation of plausible and testable hypotheses in this area may help to identify new treatment options in encephalopathies of extra-cranial origin. Therapeutic targets for this autistic phenotype may include: modification of diet and entero-colonic microbial milieu in order to reduce toxin substrates, improve nutritional status and modify mucosal immunity; anti-inflammatory/immunomodulatory therapy; and specific treatment of dysmotility, focusing, for example, on the pharmacology of local opioid activity in the gut. PMID- 11929384 TI - Review article: faecal markers in the assessment of activity in inflammatory bowel disease. AB - The fundamental pathological process behind ulcerative colitis and Crohn's disease is intestinal inflammation. As the precise cause of this is not yet completely understood, current treatment strategies are aimed at reducing or eliminating the inflammation. Endoscopic examination and histological analysis of biopsy specimens remain the 'gold standard' methods for detecting and quantifying bowel inflammation; however, these techniques are costly, invasive, and repeated examinations are unpopular with patients. Disease activity questionnaires and laboratory 'inflammatory markers', although widely used, show an unreliable correlation with endoscopy and histology. New markers need to be developed to detect and quantify bowel inflammation. These would be of use diagnostically and also an aid to pharmacological treatment. PMID- 11929385 TI - Review article: an approach to Helicobacter pylori infection in the elderly. AB - The prevalence of Helicobacter pylori infection increases with age world-wide, reaching levels of 40-60% in asymptomatic elderly subjects and over 70% in elderly patients with gastroduodenal diseases. However, the percentage of H. pylori-positive elderly patients who are treated for their infection remains very low. Data are now available that demonstrate the benefit of curing H. pylori infection in elderly patients with H. pylori-associated peptic ulcer disease and severe chronic gastritis. Furthermore, the cure of H. pylori may prevent the progression of intestinal metaplasia and gastric atrophy. New studies are needed to clarify the role of eradication in elderly patients with non-ulcer dyspepsia and gastro-oesophageal reflux disease and in those who use non-steroidal anti inflammatory drugs. H. pylori infection may be easily diagnosed by histological evaluation, rapid urease test or culture performed on gastric biopsies taken during endoscopy. However, the biopsy site must be carefully selected in elderly patients. For non-invasive monitoring of H. pylori infection after treatment, the 13C-urea breath test has significantly higher accuracy than serology in the elderly; further studies are needed to clarify the role of the H. pylori stool antigen test in old age. One-week proton pump inhibitor-based triple therapy regimens, including clarithromycin, amoxicillin and/or nitroimidazoles, are highly effective and well tolerated in elderly patients. Low doses of both proton pump inhibitors and clarithromycin (in combination with standard doses of amoxicillin or nitroimidazoles) are sufficient. Low compliance and antibiotic resistance are the main factors related to treatment failure in old age. PMID- 11929386 TI - The efficacy of methotrexate for maintaining remission in inflammatory bowel disease. AB - BACKGROUND: The management of patients with inflammatory bowel disease who are resistant to or intolerant of azathioprine remains a challenge. Low-dose methotrexate has been shown to be effective in inducing remission in Crohn's disease. AIM: This review was conducted because there are limited long-term follow-up data during and after stopping treatment. There are also limited data on the use of methotrexate in ulcerative colitis. METHODS: The study was a retrospective review of clinical notes. Remission was defined as minimal bowel symptoms without the need for oral steroids for 3 months. Relapse was defined as bowel symptoms that required steroid treatment or surgery. RESULTS: Seventy patients were reviewed; 48 had Crohn's disease and 22 had ulcerative colitis. The mean duration of treatment was 17.1 months; the mean maintenance dose was 20 mg weekly. Remission was achieved in 34 of 55 patients who completed more than 3 months of treatment (62%). Life-table analysis showed that the chances of remaining in remission at 12, 24 and 36 months (if treatment was continued) were 90%, 73% and 51%, respectively. The chances of remaining in remission after stopping treatment at 6, 12 and 18 months were 42%, 21% and 16%, respectively. The dose of methotrexate (mg/kg) was associated with the induction of remission (P=0.02). Treatment was equally effective for Crohn's disease and ulcerative colitis. CONCLUSIONS: Maintenance methotrexate treatment gives acceptable remission rates for treatment periods up to 3 years. After stopping treatment, relapse is frequent and occurs early (usually within 1 year). PMID- 11929387 TI - A pilot study of treatment of active ulcerative colitis with natalizumab, a humanized monoclonal antibody to alpha-4 integrin. AB - BACKGROUND: Alpha-4 integrins facilitate leucocyte migration across vascular endothelium. AIM: To assess the safety and efficacy of natalizumab (Antegren), a humanized antibody to alpha-4 integrin, in patients with active ulcerative colitis. METHODS: Ten patients with active ulcerative colitis, defined by a Powell-Tuck activity score > 4, received a single 3 mg/kg natalizumab infusion. The primary end-point was the change in Powell-Tuck score at 2 weeks post infusion. RESULTS: Significant decreases in the median Powell-Tuck score were observed at 2 and 4 weeks post-infusion (7.5 and 6, respectively) compared to the median baseline score (10). Five of 10 patients achieved a good clinical response at 2 weeks and one more patient by 4 weeks, defined by a Powell-Tuck score of < or = 5. Significant improvements in quality of life scores were found at week 4. Rescue medication was required by two (20%), three (30%) and eight (80%) patients by weeks 2, 4 and 8, respectively (median, 34 days; range, 8-43 days). One patient remained in remission at 12 weeks. The median C-reactive protein at 2 weeks (6 mg/L) was lower than that pre-treatment (16 mg/L). CONCLUSIONS: A single 3 mg/kg infusion of natalizumab was well tolerated by ulcerative colitis patients. The positive efficacy demonstrated in this study merits further investigation by randomized, placebo-controlled trials. PMID- 11929388 TI - Influence of acute and chronic alcohol intake on the clinical course and outcome in acetaminophen overdose. AB - BACKGROUND: Animal studies on acetaminophen toxicity suggest that chronic alcohol intake affects the outcome adversely, whereas acute alcohol intake seems protective. Few clinical data are available. METHODS: We studied 209 consecutive patients with single-dose acetaminophen overdose. The combined influence of independent variables (gender, age, dose, delay to antidote treatment, chronic and acute alcohol intake and nomogram risk group) on dependent variables (death, development of hepatic encephalopathy and biochemical liver markers) was studied using multiple or logistic regression analysis. RESULTS: Fifty-seven (27.3%) patients had chronic alcohol intake and 45 (21.5%) patients had acute alcohol intake. Forty-four (21.1%) patients developed hepatic coma and 20 (43.5%) of these patients died. Chronic alcohol intake was significantly and independently associated with the development of hepatic coma, with a lower prothrombin index, lower platelet count, higher creatinine and higher bilirubin. The relative risks for hepatic coma and death were 5.3 (95% confidence interval, 2.2-12.4) and 1.4 (95% confidence interval, 0.5-3.9), respectively, in the chronic alcohol intake group compared with the no chronic alcohol intake group. Acute alcohol intake was not significantly associated with any of the dependent variables studied. CONCLUSIONS: Chronic alcohol intake enhances acetaminophen hepatotoxicity, whereas acute alcohol intake does not affect the clinical course. PMID- 11929390 TI - Antibiotic prophylaxis prior to endoscopic retrograde cholangiopancreatography in patients with obstructive jaundice: is it worth the cost? AB - BACKGROUND: There are few published data concerning the economic impact of antibiotic prophylaxis prior to endoscopic retrograde cholangiopancreatography in the setting of biliary obstruction. AIM: To perform decision analysis to determine the costs of prophylaxis in patients undergoing endoscopic retrograde cholangiopancreatography for obstructive jaundice. METHODS: A decision analysis model was constructed. The probability of biliary sepsis, death and endoscopic retrograde cholangiopancreatography complications was obtained from the medical literature and from a retrospective analysis of our own experience. Costs were obtained from Medicare reimbursement at our institution. The strategies evaluated were endoscopic retrograde cholangiopancreatography with and without single-dose antibiotic prophylaxis. We compared the total costs, number of episodes of cholangitis and deaths associated with each strategy. RESULTS: Based on published data and the results of our retrospective analysis, the strategy of administering single-dose prophylactic antibiotics prior to endoscopic retrograde cholangiopancreatography in patients with obstructive jaundice resulted in lower total costs, fewer episodes of cholangitis and fewer deaths compared to a strategy of not administering antibiotics. The results were sensitive to the rates of cholangitis, cost of antibiotics and the cost of treating an episode of cholangitis. CONCLUSIONS: Antibiotic prophylaxis prior to endoscopic retrograde cholangiopancreatography results in fewer cases of cholangitis and is cost saving when compared to a strategy of no prophylaxis in patients with obstructive jaundice. PMID- 11929389 TI - 13C-aminopyrine breath test to evaluate severity of disease in patients with chronic hepatitis C virus infection. AB - BACKGROUND: There are few data on the use of the 13C-aminopyrine breath test to evaluate the severity of disease in patients with hepatitis C virus-related chronic liver disease, although these patients represent one of the most important problems in clinical hepatology. AIMS: To compare 13C-aminopyrine breath test results of patients with hepatitis C virus-related chronic hepatitis and Child-Pugh class A cirrhosis with those of normal subjects, and to evaluate different methods of expressing 13C-aminopyrine breath test results. METHODS: Twenty-four patients with hepatitis C virus-related chronic hepatitis and 17 patients with Child-Pugh class A cirrhosis underwent 13C-aminopyrine breath test. Breath samples were collected every 30 min up to 2 h after 13C-aminopyrine administration. 13C-Aminopyrine breath test results were expressed as a percentage of the administered dose of 13C recovered per hour (% dose/h) and the cumulative percentage of administered dose of 13C recovered over time (% dose cum). Nineteen healthy subjects served as controls. Patients with hepatitis C virus-related chronic hepatitis were divided into subgroups on the basis of histological staging and grading. RESULTS: The 13C-aminopyrine breath test result (% dose/h) at 30 min was significantly different among the three subgroups of subjects (normal subjects, 11.5 +/- 3.5; chronic hepatitis patients, 8.1 +/- 4.1; cirrhosis patients, 5.0 +/- 3.1; P < 0.0005). Moreover, the differences between chronic hepatitis and cirrhosis patients were statistically significant (P < 0.03). The fibrosis score showed a significant inverse correlation with the 13C aminopyrine breath test result (% dose/h) at 30 min (rs=- 0.409, P=0.05). The 13C aminopyrine breath test result (% dose/h) at 30 min also allowed normal subjects and chronic hepatitis patients with low (< or = 2) or high (> 2) fibrosis scores to be distinguished. The 13C-aminopyrine breath test results (% dose cum) at 30, 60 and 90 min allowed discrimination between normal subjects and chronic hepatitis and cirrhosis patients. The 13C-aminopyrine breath test result (% dose cum) was also able to distinguish between normal subjects and chronic hepatitis patients with high but not low fibrosis scores. Both 13C-aminopyrine breath test results (% dose/h and % dose cum) at 120 min allowed the differentiation between normal subjects and chronic hepatitis patients with high (> or = 6) necro inflammatory activity. CONCLUSIONS: In patients with hepatitis C virus-related chronic liver disease, the 13C-aminopyrine breath test proved to be safe and easy to perform, and was able to evaluate different degrees of liver function impairment which were partly correlated to clinical and histological evaluation. In future studies, 13C-aminopyrine breath test results should be expressed in a standardized fashion to permit comparison. PMID- 11929391 TI - There are no reliable symptoms for erosive oesophagitis and Barrett's oesophagus: endoscopic diagnosis is still essential. AB - AIMS: To evaluate the sensitivity and specificity of different symptoms in erosive reflux oesophagitis and Barrett's oesophagus. METHODS: The presence of reflux symptoms was compared between a case population of 306 patients with endoscopically determined erosive reflux oesophagitis, 235 patients with biopsy proven Barrett's oesophagus and a control population of 198 subjects without reflux disease. RESULTS: Heartburn at any time and heartburn at night represented the only two symptoms to be simultaneously sensitive and specific. Symptoms that were induced by various foods, such as fat, tomato, chocolate, citrus or spices, tended to cluster in the same sub-group of patients. Similarly, heartburn induced by exercise, lying down or bending over tended to occur in the same sub-groups. The frequency of symptoms was influenced more by the presence of mucosal erosions than by the presence of Barrett's oesophagus. Reflux symptoms occurred more frequently in the presence rather than the absence of Barrett's oesophagus, and in long segment rather than short segment of Barrett's mucosa. CONCLUSIONS: Endoscopic inspection of the oesophageal mucosa remains the only certain method by which to reliably diagnose erosive reflux oesophagitis and Barrett's oesophagus. PMID- 11929392 TI - Effect of cisapride on nocturnal transient lower oesophageal sphincter relaxations and nocturnal gastro-oesophageal reflux in patients with oesophagitis: a double-blind, placebo-controlled study. AB - AIM: To investigate the effect of cisapride, a selective 5-hydroxytryptamine-4 receptor agonist, on the frequency of nocturnal transient lower oesophageal sphincter relaxations and oesophageal acid exposure in patients with gastro oesophageal reflux disease. METHODS: In a double-blind, placebo-controlled study, 10 patients with gastro-oesophageal reflux disease (six male and four female; mean age, 54 +/- 10.4 years) were randomly assigned to 5-day treatments with cisapride, 10 mg q.d.s., or placebo, separated by a 2-day washout period before the treatment crossover. Sleep stages, lower oesophageal sphincter tone and oesophageal pH were monitored overnight at the end of each treatment regimen. Gastric emptying was assessed before treatment. RESULTS: Cisapride decreased the frequency of transient lower oesophageal sphincter relaxations during sleep (1.2 +/- 0.2/h vs. 2.7 +/- 0.5/h with placebo; P=0.004) and oesophageal acid exposure (17.2 +/- 9.9% with placebo vs. 7.2 +/- 4.2% with cisapride; P=0.4). Cisapride increased lower oesophageal sphincter tone from 12.7 +/- 2.8 mmHg with placebo to 16.9 +/- 3.9 mmHg (P=0.03), and decreased heartburn episodes and antacid consumption. All patients had normal gastric retention data over 4 h. CONCLUSIONS: In patients with gastro-oesophageal reflux disease, cisapride significantly decreased the frequency of transient lower oesophageal sphincter relaxations during sleep and increased lower oesophageal sphincter pressure without changing gastric emptying. We hypothesize, therefore, that 5 hydroxytryptamine-4 mechanisms are important in the control of transient lower oesophageal sphincter relaxations in humans. PMID- 11929393 TI - Efficacy of a motilin receptor agonist (ABT-229) for the treatment of gastro oesophageal reflux disease. AB - BACKGROUND: ABT-229 is a potent motilin agonist without significant antibiotic activity. It has been shown to improve gastric emptying in humans and to increase lower oesophageal sphincter pressure in cats. AIM: To assess the efficacy of four different doses of ABT-229 (1.25 mg, 2.5 mg, 5 mg, 10 mg b.d.) compared to placebo in the treatment of gastro-oesophageal reflux disease, and to determine its safety in patients with gastro-oesophageal reflux disease. METHODS: In a double-blind, multicentre study, 324 patients with heartburn were randomized to receive four different doses of ABT-229 or placebo for 8 weeks. The efficacy was evaluated by Patient Symptom Questionnaire, daily diary, endoscopy and global evaluation of efficacy. RESULTS: There were no statistically significant improvement scores for any of the ABT-229 treatment groups vs. the placebo group in any of the efficacy parameters. Reflux symptom scores were significantly worse after treatment in the dyspeptic group. ABT-229 appeared to be well tolerated and safe in total daily doses up to 20 mg. CONCLUSION: ABT-229 appears to have limited, if any, clinical utility in the treatment of gastro-oesophageal reflux disease. PMID- 11929394 TI - Effects of prucalopride on colonic transit, anorectal function and bowel habits in patients with chronic constipation. AB - BACKGROUND: There is a need for better tolerated drugs to normalize bowel function in chronic constipation. Prucalopride is a highly selective, specific, serotonin4 receptor agonist with enterokinetic properties. AIM: To evaluate the effects of prucalopride on bowel function, colonic transit and anorectal function in patients with chronic constipation. METHODS: Twenty-eight patients were enrolled in this double-blind, placebo-controlled, crossover study (prucalopride: 1 mg, n=12; 2 mg, n=16). Patients kept a bowel function diary. Colonic transit times and anorectal function (anal manometry, rectal sensitivity and rectal compliance) were assessed. RESULTS: Prucalopride (1 mg) compared to placebo significantly increased the mean number of spontaneous complete, spontaneous and all bowel movements per week. Prucalopride (1 mg) significantly decreased the percentage of bowel movements with hard/lumpy stools and straining and increased the urge to defecate. Prucalopride (1 and 2 mg) decreased the mean total colonic transit time by 12.0 h (prucalopride 42.8 h vs. placebo 54.8 h; P=0.074). No statistically significant effects were found in any of the anorectal function parameters. Prucalopride was well tolerated. There were no clinically relevant changes in standard safety parameters. CONCLUSIONS: Prucalopride significantly improves stool frequency and consistency, and the urge to defecate, and may decrease colonic transit times in patients with chronic constipation. PMID- 11929395 TI - Effect of octreotide on fluid absorption and secretion by the normal human jejunum and ileum in vivo. AB - BACKGROUND: We hypothesized that part of the non-specific antidiarrhoeal effect of octreotide is mediated by a proabsorptive or antisecretory effect on small intestinal active electrolyte transport. METHODS: To measure the effect of octreotide on net absorption, the jejunum and ileum of normal human subjects were perfused with a balanced electrolyte solution; to measure the effect of octreotide on normal active chloride secretion, the jejunum was perfused with a bicarbonate-free solution. RESULTS: During perfusion of a balanced electrolyte solution, octreotide increased basal net fluid absorption in the jejunum and ileum by about 40 mL/h per 30 cm. In the jejunum, octreotide markedly inhibited basal and sham feeding-stimulated active chloride secretion and inhibited water secretion by 28 and 51 mL/h per 30 cm, respectively. CONCLUSIONS: Octreotide causes an increase in the net epithelial cell absorption rate of a balanced electrolyte solution in the normal jejunum and ileum. In the jejunum, this proabsorptive effect is mediated mainly by the reduction of normal active electrolyte secretion, rather than by stimulation of normal active electrolyte absorption. These results support the hypothesis that part of the antidiarrhoeal action of octreotide is due to its effects on active electrolyte transport mechanisms by normal epithelial cells of the small intestine. PMID- 11929396 TI - Helicobacter pylori increases the risk of upper gastrointestinal bleeding in patients taking low-dose aspirin. AB - AIM: To evaluate the role of Helicobacter pylori infection and other clinical factors in the risk of upper gastrointestinal bleeding in patients taking low dose aspirin. SUBJECTS AND METHODS: A case-control study was carried out of consecutive current users of low-dose aspirin admitted because of upper gastrointestinal bleeding. Within a cohort of 695 patients with upper gastrointestinal bleeding, 98 patients had taken low-dose aspirin and no other non-steroidal anti-inflammatory drug. Controls were 147 low-dose aspirin users without upper gastrointestinal bleeding of similar age, sex and extent of aspirin use as cases. H. pylori infection was determined by CagA/VacA serology and 13C urea breath test in all cases and controls. Adjusted odds ratios (OR) are provided. RESULTS: H. pylori infection was identified as an independent risk factor of upper gastrointestinal bleeding in this population (OR, 4.7; 95% confidence interval (95% CI), 2.0-10.9), but the presence of CagA-positive serology was not. Other risk factors identified were a previous ulcer history (OR, 15.2; 95% CI, 3.8-60.1), alcohol use (OR, 4.2; 95% CI, 1.7-10.4) and use of calcium channel blockers (OR, 2.54; 95% CI, 1.25-5.14). Antisecretory therapy (OR, 0.1; 95% CI, 0.02-0.3) and nitrovasodilators (OR, 0.2; 95% CI, 0.1-0.6) decreased the risk of bleeding. CONCLUSIONS: H. pylori infection is a risk factor for upper gastrointestinal bleeding in low-dose aspirin users, which might have therapeutic implications in high-risk patients. PMID- 11929397 TI - The 13C-urea breath test to detect Helicobacter pylori infection: a validated simple methodology with 50 mg 13C-urea. AB - BACKGROUND: Several modifications of the 13C-urea breath test have been published. For reasons of cost efficiency and practicability, the urea dose and measurement duration should be reduced while still maintaining excellent diagnostic accuracy. AIMS: To establish a validated simple protocol for the urea breath test with 50 mg 13C-urea and to compare this protocol with the conventional urea breath test with 100 mg 13C-urea. METHODS: Conventional urea breath test with 100 mg 13C-urea was performed on 152 dyspeptic patients. Full cream cow's milk was used as the test meal. Breath tests were repeated using 50 mg 13C-urea and the breath samples were collected at baseline and at 10 (protocol t10), 15 (protocol t15) and 30 min (protocol t30). Helicobacter pylori status was assessed by rapid urease test, histology and conventional urea breath test with 100 mg 13C-urea. RESULTS: With protocol t15, the best combination of sensitivity (99.1%), specificity (97.3%) and accuracy (98.7%) was obtained with a cut-off of 2.5 per thousand. There was an extremely high correlation coefficient between the three protocols and conventional 13C-urea breath test (all P < 0.001). CONCLUSIONS: A urea breath test with 50 mg 13C-urea using a simple test meal and a 15-min sampling interval with a low cut-off seems to be cost-effective and convenient. In a well-standardized laboratory, this modification is not associated with any loss of diagnostic accuracy. PMID- 11929399 TI - Effect of Helicobacter pylori infection and its eradication on nutrition. AB - AIMS: To investigate the effects of Helicobacter pylori infection and eradication on nutrition. METHODS: The body weight, height, blood pressure, gastric juice pH and fasting serum levels of glucose, total protein, albumin, total cholesterol and triglyceride were measured in H. pylori-positive and H. pylori-negative subjects, and the effect of eradication of H. pylori on these parameters was determined. The development of gastro-oesophageal reflux disease after treatment was also examined. Eight patients underwent a pancreatic function test before and after H. pylori eradication therapy. RESULTS: The incidence of hypoproteinaemia in H. pylori-positive subjects was significantly higher than that in H. pylori negative subjects. After eradication of H. pylori, the gastric juice pH values were significantly decreased, and the body weight and serum levels of total cholesterol, total protein and albumin were significantly increased. The incidence of hyperlipidaemia significantly increased and that of hypoproteinaemia significantly decreased in the group with eradication. Pancreatic function improved significantly after eradication of H. pylori. No significant changes in these parameters were observed in the group without eradication. Obese patients had a higher risk of the development of gastro-oesophageal reflux disease after eradication of H. pylori infection. CONCLUSIONS: The eradication of H. pylori appears to improve some nutritional parameters. PMID- 11929398 TI - One-week omeprazole, furazolidone and amoxicillin rescue therapy after failure of Helicobacter pylori eradication with standard triple therapies. AB - AIM: To test the efficacy of omeprazole, furazolidone and amoxicillin triple therapy for the treatment of Helicobacter pylori infection after failure of standard first-line therapy recommended by the Asia-Pacific Consensus on the management of H. pylori infection. METHODS: Patients with failed H. pylori eradication received omeprazole, 20 mg, furazolidone, 100 mg, and amoxicillin, 1 g, all twice daily for 1 week. Endoscopy (CLO test, histology and culture) was performed before treatment. Post-treatment H. pylori status was determined by 13C urea breath test 6 weeks later. RESULTS: Fifty patients were recruited. Resistance to metronidazole, clarithromycin and both drugs was in the range of 50 64%, 60-75% and 40-50%, respectively, after failure of first-line therapy. Amoxicillin resistance was not found. The intention-to-treat and per protocol H. pylori eradication rates were 52% and 53%, respectively. Patients with double resistance to metronidazole and clarithromycin showed the lowest eradication rate (38%), which was significantly lower than that of patients with sensitive strains (88%). Side-effects were minimal and compliance was excellent (98%). CONCLUSIONS: One-week omeprazole, furazolidone and amoxicillin rescue therapy achieved a high eradication rate in strains sensitive to metronidazole and clarithromycin. This is a cheap and safe rescue regimen when guided by pre-treatment sensitivity testing. PMID- 11929400 TI - Serum pepsinogen and gastrin levels in HIV-positive patients: relationship with CD4+ cell count and Helicobacter pylori infection. AB - BACKGROUND: The relationship between serum parameters of gastric function and Helicobacter pylori infection in human immunodeficiency virus (HIV)-positive patients is almost unknown. AIMS: To investigate in HIV-infected patients: (i) the relationship between serum gastrin and serum pepsinogens over the progressive phases of HIV-related disease; (ii) the impact of H. pylori infection on gastrin and pepsinogen serum levels and its relation to antral histology; (iii) the prevalence of parietal cell autoantibodies. METHODS: Fifty-nine HIV-positive patients were studied by upper endoscopy plus gastric antral biopsy. Serum samples were tested for gastrin, pepsinogen A, pepsinogen C and parietal cell autoantibodies. RESULTS: In patients without overt acquired immunodeficiency syndrome (AIDS), or with a CD4+ count of > 100 x 10(6) cells/L, mean serum levels of gastrin and pepsinogen C were higher than in subjects with AIDS or with a CD4+ count of < 100 x 10(6) cells/L (P < 0.01). Only one patient was found to be positive for parietal cell autoantibodies. H. pylori infection was associated with increased values of gastrin and pepsinogen C only in HIV-positive patients without AIDS or with a CD4+ count of > 100 x 10(6) cells/L. Atrophy was more frequent in patients with overt AIDS than in those without overt AIDS (57% vs. 33%, P=N.S.), and/or in patients with a CD4+ count of < 100 x 10(6) cells/L than in those with a CD4+ count of > 100 x 10(6) cells/L (62% vs. 26%, P < 0.05). CONCLUSIONS: HIV-positive patients without overt AIDS have increased serum levels of gastrin and pepsinogen C compared with HIV-positive patients with overt AIDS. PMID- 11929401 TI - An evaluation of a rapid urine test for the diagnosis of Helicobacter pylori infection in the Chinese population. AB - BACKGROUND: A new rapid urine test was developed to detect anti-Helicobacter pylori antibody in urine using the principle of immunochromatography. The accuracy of this test in the Chinese population remains to be defined. AIM: To evaluate a new rapid urine test for the diagnosis of H. pylori infection in the Chinese population. METHODS: Eligible patients without previous treatment of H. pylori were recruited. In-house rapid urease test and histology were used as the gold standard. The rapid urine test (RAPIRUN H. pylori antibody) was performed and the results were compared with the gold standard. RESULTS: One hundred and twenty-three patients were eligible for analysis and 61 (50%) were H. pylori positive by the gold standard. The rapid urine test showed a sensitivity, specificity, positive predictive value, negative predictive value and accuracy of 96.7%, 95.2%, 95.2%, 96.7% and 95.9%, respectively. Results were obtained within 20 min and no special laboratory support was required. CONCLUSIONS: The rapid urine test by immunochromatography is highly accurate for the diagnosis of H. pylori infection in untreated Chinese patients. PMID- 11929403 TI - Dose-dependent control of intragastric pH by pantoprazole, 10, 20 or 40 mg, in healthy volunteers. AB - BACKGROUND: Proton pump inhibitors have emerged as the most effective class of drugs for the treatment of gastro-oesophageal reflux. Pantoprazole is a proton pump inhibitor that has demonstrated high clinical efficacy. AIM: To evaluate the effect of once-daily doses of pantoprazole, 10, 20 and 40 mg, on gastric acidity in healthy volunteers. METHODS: Thirty-six subjects received pantoprazole in a three-way crossover design study. Ambulatory 24-h intragastric pH and distal oesophageal pH were monitored at baseline and on the last day of each treatment period. The measured endpoints were the median intragastric and oesophageal pH, the percentage of time the intragastric pH < 4 and oesophageal pH < 4 and the area under the curve for gastric acidity over 24 h. Safety was evaluated by incidence and severity of adverse events. RESULTS: Pantoprazole demonstrated a linear dose- dependent suppression of gastric acidity over the dose range 10-40 mg. The dose of 40 mg demonstrated a significantly greater response than the lower doses, particularly at night. All pantoprazole doses were well tolerated. CONCLUSIONS: Pantoprazole demonstrates a dose-related effect in the range 10-40 mg once daily. The once-daily dose of 40 mg provides the highest and most consistent control of gastric pH, especially at night. PMID- 11929402 TI - Dyspepsia tolerability from the patients' perspective: a comparison of celecoxib with diclofenac. AB - AIM: To compare celecoxib (800 mg/day, n=1997) with diclofenac (150 mg/day, n=1996) on dyspepsia-related tolerability. METHODS: In one of the two protocols comprising the Celecoxib Long-Term Arthritis Safety Study, a randomized double blind trial, patients completed the Severity of Dyspepsia Assessment Questionnaire at baseline and at weeks 4, 13, 26 and 52 for the following three scales: Pain Intensity, Non-Pain Symptoms and Satisfaction with Dyspepsia-Related Health. RESULTS: For the Pain Intensity scale, patients given diclofenac had significantly higher (worsening dyspepsia) mean changes, defined as follow-up minus baseline, than patients given celecoxib (P < 0.001, at all assessments). The mean changes in the Pain Intensity scale (scale, 2-47; higher score is higher pain intensity) were 0.99 (95% confidence interval (CI): 0.50, 1.48) for celecoxib and 2.76 (95% CI: 2.28, 3.25) for diclofenac at 4 weeks. Satisfaction was superior with celecoxib at all assessments (P < 0.001). At 4 weeks, the mean changes in the Satisfaction scale (scale, 7-35; higher score is higher satisfaction) were 0.02 (95% CI: - 0.26, 0.29) for celecoxib and - 0.72 (95% CI: 1.00, - 0.45) for diclofenac. Diclofenac patients had significantly higher Non Pain Symptoms at 4 weeks (P=0.005). CONCLUSIONS: Celecoxib, at two to four times the recommended dose, demonstrated a superior dyspepsia-related tolerability and satisfaction compared with standard dosages of diclofenac. PMID- 11929404 TI - Comparison of lansoprazole and famotidine for gastric acid inhibition during the daytime and night-time in different CYP2C19 genotype groups. AB - BACKGROUND: The acid inhibitory effect of lansoprazole depends on the S mephenytoin 4'-hydroxylase (CYP2C19) genotype status. The effect of famotidine is independent of this genotype. AIM: To investigate the acid inhibitory effects of lansoprazole vs. famotidine during the daytime and night-time with reference to different CYP2C19 genotypes. METHODS: Fifteen healthy volunteers were given 20 mg famotidine twice a day or 30 mg lansoprazole once a day for 8 days. On post-dose day 8, 24-h intragastric pH monitoring was performed. RESULTS: During the daytime, the intragastric pH with lansoprazole was significantly higher than that with famotidine in the heterozygous extensive metabolizer group, whereas no significant difference was observed in the homozygous extensive metabolizer group. During the night-time, the intragastric pH with famotidine was quite similar to that with lansoprazole in the heterozygous extensive metabolizer and poor metabolizer groups. However, during the night-time, the intragastric pH with famotidine was significantly higher than that with lansoprazole in the homozygous extensive metabolizer group. CONCLUSIONS: An insufficient acid inhibition by lansoprazole during the night-time in the homozygous extensive metabolizer group could be compensated for by famotidine. CYP2C19 genotype testing appears to be useful for predicting the optimal acid inhibitory drug treatment collated with circadian intragastric pH change. PMID- 11929405 TI - Inhibition of Helicobacter pylori-induced cyclo-oxygenase-2 aggravates NSAID caused gastric damage in Mongolian gerbils. AB - BACKGROUND: The effect of Helicobacter pylori infection on non-steroidal anti inflammatory drug-induced gastric mucosal injury is controversial. AIM: To examine the effect of the interaction between H. pylori and non-steroidal anti inflammatory drugs on gastric mucosal injury. METHODS: Mongolian gerbils infected with H. pylori were treated with indometacin at 8 mg/kg for 2 days or 7 days. Mucosal damage was assessed by macroscopic and histological examination, and myeloperoxidase activity was measured as an index of neutrophil infiltration. The expression levels of cyclo-oxygenase proteins were determined by Western blot analysis and cyclo-oxygenase activity. RESULTS: A 2-day course of indometacin did not cause an increase in gastric damage in H. pylori-infected Mongolian gerbils compared to uninfected gerbils, while a 7-day course of indometacin caused additive gastric damage in H. pylori-infected animals. H. pylori infection induced cyclo-oxygenase-2 expression in the stomach. Treatment with indometacin for 2 days did not significantly affect cyclo-oxygenase activity in H. pylori infected animals, while treatment for 7 days inhibited both cyclo-oxygenase-1 and cyclo-oxygenase-2 activities. Pre-treatment with a selective cyclo-oxygenase-2 inhibitor aggravated mucosal injury in H. pylori-infected animals treated or not treated with indometacin for 2 days. CONCLUSIONS: Our results suggest that cyclo oxygenase-2 protein induced by H. pylori infection may be involved in the defence of the gastric mucosa against damage caused by non-steroidal anti-inflammatory drugs. Therefore, inhibition of cyclo-oxygenase-2 activity may enhance non steroidal anti-inflammatory drug-caused gastric damage in H. pylori-infected animals. PMID- 11929406 TI - T-cell reactivity in neonates: influence of environmental and genetic factors. PMID- 11929407 TI - Has in vitro allergenicity testing any clinical relevance? PMID- 11929408 TI - Statistical issues--significantly important in medical research. PMID- 11929409 TI - Evaluation of the residual antigenicity and allergenicity of cow's milk substitutes by in vitro tests. AB - BACKGROUND: This study aimed the to investigate presence of residual allergenic cow's milk proteins (CMP) in some milk substitutes employed in the treatment of cow's milk allergy (CMA). These allergens may interfere with the treatment, and elicit allergic reactions in sensitized individuals. METHODS: The protein composition of the different extracts was evaluated by Lowry's method and tricine SDS-PAGE. Different immunoenzymatic methods were used (ELISA, EAST and immunoblotting) to quantify total serum IgE and specific serum IgE, as well as to detect the presence of antigenic and allergenic components. RESULTS: The results showed a higher protein content in mammalian milks (cow, sheep, mare, goat, and human) than in hydrolyzed substitutes (partially or extensively hydrolyzed casein or whey proteins). Residual native, processed, or contaminant polypeptides have been identified in the moderate hydrolysates, whereas extensive hydrolysates did not show the presence of residual components by immunoblotting. However, specific antibodies with capacity to bind to peptides have been detected by EAST and ELISA, suggesting that extensive hydrolysates contain residual peptides that preserve immunoreactive epitopes. We were unable to demonstrate either residual antigenicity or allergenicity in an amino-acid-based formula. CONCLUSIONS: Immunoenzymatic methods were used to detect the presence of cross-reactive components in mammalian milks. Residual allergenic components from cow's milk could be identified in both the moderate and extensive hydrolysates analyzed. This information may be relevant to the treatment of CMA. PMID- 11929410 TI - Privet pollen (Ligustrum sp.) as potential cause of pollinosis in the city of Cordoba, south-west Spain. AB - BACKGROUND: Privet pollen rarely accounts for more than 1% of the annual total of daily pollen concentrations measured in a city; however in areas where these trees are widely used as ornamentals the amounts collected may be high enough to cause allergy symptoms. METHODS: Air samples taken with volumetric particle samplers Lanzoni VPPS 1000 (Lanzoni s.r.l., Bologna, Italy) show that there are differences in privet pollen concentrations measured in neighbourhoods with a high incidence of privet trees and in those taken at some distance from the source of emission. RESULTS: The results suggest that differences are due to the short dispersal range of the pollen grains once released from the plant, resulting from both the entomophilous nature of the plant and the large size of the pollen grains. Urban design, moreover, may play an important role in impeding pollen grain dispersion if the air cannot flow freely through long, narrow avenues. Another important consideration is that the last stages of the flowering period of privet overlaps with the flowering period of olive trees, the main allergen in the area. The fact that the two pollen types share common allergens means that there may be a cross-reaction between olive tree pollen and privet pollen. CONCLUSIONS: Privet pollen should be considered as a potential causative agent of local allergy problems in areas where its presence is extensive and is in combination with other allergens. PMID- 11929411 TI - IgE-mediated allergy to corn: a 50 kDa protein, belonging to the Reduced Soluble Proteins, is a major allergen. AB - BACKGROUND: Although corn is often cited as an allergenic food, very few studies have been devoted to the identification of corn allergens and corn allergy has been rarely confirmed by double-blind, placebo-controlled food challenge (DBPCFC). Recently, Pastorello et al. (1) identified some salt-soluble IgE binding proteins of corn flour as potential allergens. One of these, corresponding to corn Lipid Transfer Protein (LTP), appeared to be the major one. The aim of this study was to verify the clinical significance of the skin prick test (SPT) and CAP-FEIA CAP-System IgE fluozoenzyme immunosorbent assay (Pharmacia Diagnostic, Uppsala, Sweden) positivities to corn and to identify the presence of IgE-binding proteins in the corn flour salt-insoluble protein fractions (comprising up to 96% of the total protein) using sera of patients with DBPCFC-documented food allergy to corn. In addition the effect of cooking and proteolytic digestion on the corn allergens was investigated. METHODS: Sixteen subjects with SPT and CAP-FEIA positivities to corn flour were examined. Only six of them complained of suffering from urticaria and/or other symptoms after ingestion of corn-based foods. The patients were food challenged with cooked corn flour (polenta). IgE-binding proteins were detected by immunoblotting. The digestibility of the IgE-binding proteins was examined during a pepsin attack followed by a pancreatin digestion performed on a cooked corn flour sample. RESULTS: Oral challenge was positive only for six patients with symptoms after ingestion of corn. A 50 kDa protein, belonging to the corn Reduced Soluble Protein (RSP) fraction was recognized by the serum IgE of all the DBPCFC-positive subjects and resulted to be resistant to both heating and peptic/pancreatic digestion. SPT with the purified RSP fraction gave positive results for all of the DBPCFC-positive patients examined. CONCLUSIONS: SPT and CAP-FEIA positivities to corn flour had no clinical significance for most of the patients and food allergy to corn has to be proved by DBPCFC. A salt-unextractable protein of 50 kDa, belonging to the RSP fraction, represents a potential allergen in food hypersensitivity to corn because of its stability to cooking and digestion. PMID- 11929412 TI - The score for allergic rhinitis (SFAR): a simple and valid assessment method in population studies. AB - BACKGROUND: No validated assessment of allergic rhinitis (AR) is presently available that can be used in population studies in the absence of medical diagnosis and of objective measurements of allergy. To compensate for this lack, a quantitative Score For Allergic Rhinitis (SFAR) ranging between 0 and 16 has been developed by experts. METHODS: The SFAR, encompassing eight features of AR, was validated in three different ways: 1) among 269 outpatients taking the specialist's diagnosis of AR and skin prick tests (SPT) positivity as a gold standard (diagnosis validation); 2) using psychometric methods (internal validation); and 3) in a random population-based sample of 3001 individuals by telephone interview (population acceptability). RESULTS: A SFAR value > or = 7 allowed satisfactory discrimination between the outpatients with AR from those without (sensitivity = 74% [95% confidence interval CI: 0.69,0.79], specificity = 83% [0.79, 0.87], positive predictive value = 84% [0.80, 0.88], negative predictive value = 74% [0.69, 0.79] and Youden's index = 0.57, respectively). Internal consistency of the score was also high (among others, Cronbach's alpha coefficient = 0.79). On average, it took only 3 min for the individuals interviewed on the phone to complete the questionnaire, the questions of which were well understood. Among these subjects, the prevalence of AR was 21% [95% CI: 19.5%, 22.5%], which is comparable to other determinations in France. CONCLUSIONS: The newly a priori proposed Score For Allergic Rhinitis (SFAR) is easy to use and can be useful to estimate prevalence and to study causation of AR in population settings. PMID- 11929413 TI - Asthma and allergy: the significance of chronic conditions for individual health behaviour. AB - BACKGROUND: In health economics, health is regarded as part of an individual's human capital. As such it depreciates over time, and investments in health are made in order to keep the stock of health capital at the desired level. Using this framework for analysis of health-related behaviour and Swedish panel data, we examined whether the presence of asthma or allergy affects perceived health and investments in health. METHODS: A set of panel data for approximately 3800 individuals interviewed repeatedly in 1980/81, 1988/89, and 1996/97 was created from the Swedish biannual survey of living conditions. Self-assessed health was chosen as the indicator of health capital and the reported number of sick days as the indicator of health investment. The presence of asthma or allergy, age, wage rate, wealth, marital status, number of children, exercise and smoking habits, gender, and geographic location of household were all chosen as explanatory variables. An ordered probit model was estimated for the health equation and a Poisson model for the investment equation. RESULTS: We found that both asthmatics and those who suffer from allergy invested more in their health than the general population. We also found that asthmatics reported significantly lower self assessed health than the general population, while those who suffered from allergy did not differ significantly from the general population regarding their self-assessed health. CONCLUSION: The human capital approach was found suitable for studying the impact of asthma and allergy on individual health behaviour. Health policy measures, which reduce the individual's costs of investing in his or her health, would improve health levels. Because asthmatics were found less healthy than those suffering from allergy, the potential gains would be larger for patients with asthma than for patients with allergy. The issue of whether this would be a cost-effective policy or not would require a different design and, hence, could not be solved within the present study. PMID- 11929414 TI - Factors associated with asthma in Albania. AB - BACKGROUND: This study was undertaken to see whether asthma in Albania (where it is particularly uncommon) is associated with certain 'western' characteristics, and with dietary and lifestyle factors that affect its occurrence elsewhere. METHODS: A screening survey was conducted among 2653 persons aged 20-44 years. A more detailed enquiry was conducted among a random subsample plus all others whose response indicated possible asthma. Three groups were compared: 495 probable nonasthmatics ('nonsymptomatics'); 106 with asthma-like symptoms and positive skin tests ('possible allergic asthmatics'); and 116 with similar symptoms and negative skin tests ('other symptomatics'). RESULTS: The possible allergic asthmatics were less likely than the nonsymptomatics to have attended school or nursery under the age of 5 years or to have shared a bedroom with an older child before that age. Continued education after the age of 18 years also appeared to be protective. Allowing for other factors, a strong negative relation was found with fruit and vegetable consumption, and weak positive relations with prepackaged food and fizzy drinks. Symptoms in skin prick test-negative subjects were associated with personal and parental smoking habit. CONCLUSIONS: This survey provides limited evidence linking asthma with aspects of western lifestyle. It supports the hypothesis that opportunities for infection during early childhood and the consumption of fruit and vegetables protect against asthma. PMID- 11929415 TI - T cell reactivity in neonates from an East and a West German city--results of the LISA study. AB - BACKGROUND: Within an ongoing birth cohort study (LISA) the cytokine production of cord blood T cells was compared between neonates from Leipzig (East Germany) and Munich (West Germany). The aim of this study was to analyse regional differences and influencing factors of the immune status. METHODS: Cytokine production was measured in a randomly selected subgroup of 158 children from the LISA (Life style - Immune system - Allergy) cohort by intracellular cytokine staining. Information on family "atopy" history (FAH) and home characteristics was obtained from questionnaires. RESULTS: Reduced numbers of interferon-gamma (IFN-gamma) and tumor necrosis factor-alpha (TNF-alpha) producing T cells were found in association with biparental FAH and housing renovation during pregnancy. In addition, cytokine production was influenced by season. In Munich, the frequency of biparental FAH and of renovation measures during pregnancy was significantly higher as compared to Leipzig. Neonates from Munich showed significantly decreased amounts of IFN-gamma and TNF-alpha and elevated levels of interleukin-4 (IL-4) producing T cells. Differences in cytokine production between Munich and Leipzig were influenced by season (IL-4) and housing renovation (IFN-gamma, TNF-alpha). CONCLUSIONS: Since differences in the T cell cytokine production of neonates in Munich and Leipzig are independent from FAH our findings may provide evidence for the impact of environmental factors upon the fetal immune system. PMID- 11929417 TI - Wipe test for the detection of indoor allergens. AB - BACKGROUND: The determination of house dust mite and cat allergens is a prerequisite for qualified assessment of exposure risk indoors. We describe the validation of integrated wipe tests comprising a porous sampling pin for the collection of settled house dust and an immunochemical test set-up for specific analysis of dust associated allergens. METHODS: Wipe tests comprised an immunochromatographic test system and a porous sampling pin for collecting dust by manually wiping a surface. Two different wipe tests were developed with specificity to cat allergen (Fel d 1) and to dust mite allergen (Der p 2/f 2). One-hundred and fifty on-site determinations for each of the allergens in 50 different households completed the field study to validate the analytical system. Reference analysis was based on collection of dust with a vacuum cleaner and analysis of dust associated allergens by ELISA. RESULTS: The wipe tests were found to produce semiquantitative results (class 0-3) on-site within 10 min of analysis time. For dust mite (cat) allergen the median of class 0-3 was 119 (263), 812 (844), 2708 (5670) and 8000 (56 800) ng/g dust, respectively. With respect to cut-off levels for dust mite (cat) allergens the clinical sensitivity was 62% (79%) and the clinical specificity was 94% (92%). CONCLUSIONS: The wipe tests proved to be a reliable tool for indoor allergen exposure screening, suitable for use even by untrained persons. This may have an impact on compliance with allergen avoidance measures. PMID- 11929416 TI - Predictors of high house dust mite allergen concentrations in residential homes in Sydney. AB - BACKGROUND: In parts of coastal Australia, house dust mite allergen concentrations in homes are often very high with at least 80% of homes in Sydney exceeding concentrations of 10 microg of allergen per gram of fine dust. In this study, we report the relation between characteristics of the home environment and house dust mite allergen concentrations at three sites in Sydney homes. METHODS: A total of 616 families were recruited as part of the Childhood Asthma Prevention Study (CAPS). Information about the home environment and structural aspects of the home was collected using a questionnaire. Samples of dust were collected from the parents' bed, the bedroom floor and the living room floor and assayed for Der p 1. RESULTS: A total of 68% of participants' beds, 65% of bedroom floors and 56% of living room floors had Der p 1 concentrations above 10 microg/g, with the highest concentrations of allergen in the bed. The most significant predictor of high Der p 1 concentrations in the bed and floors was the age of the home. We also found that beds with mattresses over two years old and with woollen or synthetic blankets or synthetic quilts had higher Der p 1 concentrations. Carpeted floors had higher Der p 1 concentrations than hard floors. CONCLUSION: The finding that high Der p 1 allergen concentrations in homes with carpets and older mattresses indicates that control strategies directed at these sources are likely to be effective in reducing exposure. Alternatives such as the use of house dust mite impermeable mattress encasings on older mattresses may also be effective in reducing exposure. PMID- 11929418 TI - Cross-reactivity between Platanus pollen and vegetables. AB - BACKGROUND: Several associations have been described between tree and plant pollens and certain foods. The objective of this study is to verify whether there is cross-reactivity between Platanus pollen and vegetable origin foods. METHODS: We selected 56 patients allergic to vegetable foods and subjected them to cutaneous tests with aeroallergens and vegetable foods. A statistical analysis was performed to evaluate the association of Platanus pollen with foods and with other aeroallergens. Later, a specific IgE determination was performed as well as a RAST (radioallergosorbent) inhibition experiment, to verify the existence of cross-reactivity in vitro. RESULTS: In the cutaneous tests we found a positive correlation between Platanus pollen and hazelnut, peanut, banana and celery. The results of the RAST inhibition experiment indicate an important cross-reactivity between the pollen of Platanus acerifolia and hazelnut and banana fruit, and an intermediate cross-reactivity with celery and peanut. CONCLUSION: We have described an association between the pollen of the Platanus tree and some vegetable foods such as hazelnut, banana, peanut and celery. This association could be explained by the in vitro IgE cross-reactivity detected. PMID- 11929419 TI - Rhinitis: do diagnostic criteria affect the prevalence and treatment? AB - BACKGROUND: Rhinitis is one of the world's most common health problems. Diagnostic criteria used in community surveys may affect reported prevalence and treatment. METHODS: A proportionately stratified random sample study was performed to investigate the prevalence, comorbidities and management of community-based patients with rhinitis in the tropical urban city of Singapore. RESULTS: The prevalence of at least one, two, three, or four nasal symptoms on most days during the past year in our study population was 25.5%, 13.1%, 6.5%, and 3.0%, respectively. Based on the definition of 'rhinitis' by the International Consensus Report (ICR), the prevalence was 13.1% in Singapore. There was significantly higher prevalence of self-reported allergy, asthma, and common cold/influenza-like illness among the rhinitis group. In the 53% of rhinitis subjects seeking for medical help, 71% visited a primary care physician and 20% an otolaryngologist. Treatments as reported by patients were decongestants (topical or oral) 27%, antibiotics 12%, antihistamines 6%, nasal steroids 3%, surgery 2%, traditional methods 28%, and 22% did not know what medication they had. Subjects considered the effectiveness of treatment unsatisfactory because the majority of them had only partial or no relief with any treatment. CONCLUSIONS: The standardization of the definition of rhinitis in epidemiological studies is of crucial importance, especially when comparing the prevalence between studies. Appropriate patient education by physicians with a good understanding of the nature of rhinitis and the available treatment options (e.g. evidenced-based efficacy, safety, and a good cost-benefit ratio) will maximize patient compliance and treatment outcomes. PMID- 11929420 TI - Siberian hamster: a new indoor source of allergic sensitization and respiratory disease. AB - BACKGROUND: The identification of 'unknown' allergic sensitizations may determine the prognosis and treatment of patients with respiratory airway disease. Currently, the presence in homes of 'exotic' animals as pets is increasing. In this article the Siberian hamster or dwarf hamster (Phodopus sungorus) was identified as a new indoor source of aeroallergens and respiratory disease. METHODS: The subjects were six outpatients who were treated for asthma and rhinitis. Siberian hamster hair extract was prepared with a standard wt/vol method, and patients were skin-prick tested with the extract. Serum-specific immunoglobulin (Ig)E against the Siberian hamster, common hamster (Cricetus cricetus) and golden hamster (Mesocricetus auratus) was determined. IgE immunoblotting was also performed for all six sera. Specific bronchial challenge was carried out with the Siberian hamster extract. RESULTS: Skin prick tests (SPT) with the Siberian hamster extract, and specific IgE-antibodies against Siberian hamster, were strongly positive in all six patients. Determinations of specific IgE-antibodies against C. cricetus and M. auratus were negative in all patients. IgE-immunoblotting of the sera revealed two IgE-binding fractions (MW 18 and 32 kDa) in five of the six sera. Specific bronchial provocation tests resulted in early asthmatic responses in the two patients who were challenged. CONCLUSIONS: The present study reveals the Siberian hamster to be able to induce both sensitization and disease, and this species of hamster should be taken into consideration as a cause of respiratory disease in exposed subjects. A noteworthy finding was the lack of sensitization in our patients to common hamster allergens (M. auratus and C. cricetus) that are usually tested when hamster allergy is suspected. PMID- 11929421 TI - Lack of association between atopic eczema/dermatitis syndrome and polymorphisms in the promoter region of RANTES and regulatory region of MCP-1. AB - BACKGROUND: Chemokines play an important role in the pathophysiology of atopic eczema/dermatitis syndrome (AEDS) and allergy. Recently polymorphisms in the promoter region of RANTES (regulated on activation normal T cell expressed and secreted) and in the gene regulatory region of MCP-1 (monocyte chemoattractant protein-1) have been found, which increase the expression of these chemokines. The - 403A allele of the RANTES promoter region was found associated with AEDS in German children. We investigated whether the presence of these polymorphisms was associated with AEDS or allergy in Hungarian children. METHODS: One hundred and twenty-eight children with AEDS, 102 allergic children without AEDS and 303 children of comparable ages without allergic disorders were screened for genotype with a PCR-based assay. RESULTS: There were no significant differences in the frequency of these polymorphisms, or in the distribution of genotypes between the groups. The total IgE concentration, the white blood cell count and the blood eosinophil cell count did not differ between the genotypes. CONCLUSION: In this cohort of Hungarian children there was no association between - 28G, and - 403A alleles in the RANTES promoter, - 2518G polymorphism in the distal regulatory region of the MCP-1 and AEDS, or allergy. PMID- 11929422 TI - Evaluation of Petri dish sampling for assessment of cat allergen in airborne dust. AB - BACKGROUND: Dust reservoir sampling is the most commonly used method for assessment of indirect allergen exposure. Because assessment of personal exposure using person-carried pumps is time-consuming and expensive we evaluated the Petri dish sampling method for measurement of airborne cat allergen in classrooms. METHODS: Petri dish sampling was evaluated in three study parts. Part I: by comparison between Petri dish sampling and personal air sampling in 44 classrooms with many (> or = 20%) and few (< or = 10%) cat owners and by additional Petri dish sampling in 40 pet-free homes. Part II: by sampling with duplicate Petri dishes in 28 classrooms. Part III: by sampling in three classrooms at four sampling heights during different sampling times. All samples were analyzed for cat allergen (Fel d 1) content with a monoclonal antibody two-site ELISA (enzyme linked immunosorbent assay), using signal amplification when necessary. RESULTS: There was a significant correlation between Petri dish sampling and personal air sampling (r = 0.66; P < 0.0001). Levels were five-fold higher in classes with many cat owners than in classes with few cat owners, regardless of method. A corresponding difference was found in the homes. Duplicate sample values were in fair agreement (Bland-Altman test) and were correlated (r = 0.77; P < 0.0001). Cumulative levels collected weekly in one Petri dish were lower than using five daily Petri dishes, regardless of sampling height. CONCLUSIONS: Petri dish sampling can be useful as an alternative method to personal air sampling of airborne allergens. PMID- 11929423 TI - Atopy and house dust mite sensitization as risk factors for asthma in children. AB - BACKGROUND: Recent evidence suggests that asthma is not invariably related to atopy. The aim of this study was to evaluate the frequency of atopy, asthma and sensitization to eight common allergens in a large group of children with allergic symptoms. METHODS: 1426 children referred to our Paediatric Asthma and Allergy Center because of allergic symptoms were examined. Bronchial asthma, allergic rhino-conjunctivitis, food allergy and atopic dermatitis were diagnosed with standardized methods. Atopy was diagnosed if at least one skin test was positive. RESULTS: Of the 1426 children examined, 629 (44%) were atopic and 769 (56%) were non-atopic. Asthma was diagnosed in the same proportion (i.e., 64%) of atopic and non-atopic children. However, after division into age groups, non atopic asthma was significantly more prevalent (chi2 = 8.46) in children between 0 and 3 years old (group 1). On the other hand, atopy was significantly associated with asthma only in group 3 (odds ratio 1.85). Furthermore, a significant association with asthma symptoms was found for house dust mite (HDM) in group 3 (odds ratio 4.8). CONCLUSIONS: Asthma is related to atopy in pre selected children only from the age of 7 years. House dust mite sensitization seems to be an important determinant of asthma in these "older" children. PMID- 11929424 TI - Increased levels of a TH2-type CC chemokine thymus and activation-regulated chemokine (TARC) in serum and induced sputum of asthmatics. AB - BACKGROUND: Cytokines liberated by TH2 cells play crucial roles in the pathogenesis of bronchial asthma. Recent studies have demonstrated that CC chemokine receptor (CCR)4 is preferentially expressed by TH2 cells. These facts suggest possible involvement of two CCR4-specific ligands i.e., thymus and activation-regulated chemokine (TARC) and macrophage-derived chemokine (MDC), in the pathogenesis of bronchial asthma via recruitment of TH2 cells to inflammatory sites. We investigated the levels of TARC and MDC in the serum and induced sputum of asthmatics. METHODS: The levels of TARC in the serum (46 asthmatics and 26 healthy subjects) and induced sputum (30 asthmatics and 6 healthy subjects) were measured using a highly sensitive ELISA system. The levels of eotaxin and MDC were also measured by ELISA. RESULTS: TARC, but not MDC, was significantly increased in asthmatic sera (P<0.001). Although MDC was undetectable in the sputum of most cases by our assay system, sputum TARC was significantly increased (P=0.027). CONCLUSIONS: The elevated TARC levels in asthmatics might be involved in the pathophysiology of asthma. PMID- 11929426 TI - Elevated serum leptin in AEDS. PMID- 11929427 TI - TARC in allergic disease. PMID- 11929428 TI - Risks of latex avoidance. PMID- 11929429 TI - IgE to Bet v 1/Bet v 2 in UAS. PMID- 11929430 TI - Anaphylaxis to coconut. PMID- 11929431 TI - Unusual NSAID hypersensitivity. PMID- 11929432 TI - Nonallergic anaphylaxis to pantoprazole. AB - Consider reactions to proton pump blockers in suspected food hypersensitivity PMID- 11929433 TI - A call to authors: three solid reasons to publish in the Australian and New Zealand Journal of Psychiatry. PMID- 11929435 TI - Dysregulation of the right brain: a fundamental mechanism of traumatic attachment and the psychopathogenesis of posttraumatic stress disorder. AB - OBJECTIVE: This review integrates recent advances in attachment theory, affective neuroscience, developmental stress research, and infant psychiatry in order to delineate the developmental precursors of posttraumatic stress disorder. METHOD: Existing attachment, stress physiology, trauma, and neuroscience literatures were collected using Index Medicus/Medline and Psychological Abstracts. This converging interdisciplinary data was used as a theoretical base for modelling the effects of early relational trauma on the developing central and autonomic nervous system activities that drive attachment functions. RESULTS: Current trends that integrate neuropsychiatry, infant psychiatry, and clinical psychiatry are generating more powerful models of the early genesis of a predisposition to psychiatric disorders, including PTSD. Data are presented which suggest that traumatic attachments, expressed in episodes of hyperarousal and dissociation, are imprinted into the developing limbic and autonomic nervous systems of the early maturing right brain. These enduring structural changes lead to the inefficient stress coping mechanisms that lie at the core of infant, child, and adult posttraumatic stress disorders. CONCLUSIONS: Disorganised-disoriented insecure attachment, a pattern common in infants abused in the first 2 years of life, is psychologically manifest as an inability to generate a coherent strategy for coping with relational stress. Early abuse negatively impacts the developmental trajectory of the right brain, dominant for attachment, affect regulation, and stress modulation, thereby setting a template for the coping deficits of both mind and body that characterise PTSD symptomatology. These data suggest that early intervention programs can significantly alter the intergenerational transmission of posttraumatic stress disorders. PMID- 11929434 TI - Are psychiatrists equipped for management roles in mental health services? AB - OBJECTIVES: To promote the discussion of leadership and management skills development among psychiatrists in Australia and New Zealand. METHOD: A key informant survey of fellows of the Royal Australian and New Zealand College of Psychiatrists was conducted via a semi-structured interview. This canvassed views about leadership and management issues including levels of confidence and needs for additional skills. RESULTS: There was widespread support for psychiatrists to be in management roles in mental health services; however, on entering management positions, psychiatrists often felt inadequately trained and prepared for their new role. Furthermore, many who had made the transition to management perceived a lack of support from their clinical colleagues. Clinicians appeared to believe that management was not difficult to learn and could be done by any experienced clinician. The provision of short courses and mentoring programmes is the preferred option for most psychiatrists seeking to acquire leadership and management skills. CONCLUSIONS: For psychiatrists to maximize their potential as leaders in Australian and New Zealand mental health services, greater attention to promoting the acquisition of relevant skills throughout training and in the post-fellowship years is required. Psychiatrists need to be supported and encouraged to pursue further education, training and research in this area. Failure to address this issue risks psychiatrists continuing to feel disadvantaged in management roles and hence reluctant to undertake the challenge. PMID- 11929436 TI - Magnetic resonance spectroscopy and its applications in psychiatry. AB - OBJECTIVE: This paper briefly describes neuroimaging using magnetic resonance spectroscopy (MRS) and provides a systematic review of its application to psychiatric disorders. METHOD: A literature review (Index Medicus/Medline) was carried out, as well as a review of other relevant papers and data known to the authors. RESULTS: Magnetic resonance spectroscopy is a complex and sophisticated neuroimaging technique that allows reliable and reproducible quantification of brain neurochemistry provided its limitations are respected. In some branches of medicine it is already used clinically, for instance, to diagnose tumours and in psychiatry its applications are gradually extending beyond research. Neurochemical changes have been found in a variety of brain regions in dementia, schizophrenia and affective disorders and promising discoveries have also been made in anxiety disorders. CONCLUSION: Magnetic resonance spectroscopy is a non invasive investigative technique that has provided useful insights into the biochemical basis of many neuropsychiatric disorders. It allows direct measurement, in vivo, of medication levels within the brain and has made it possible to track the neurochemical changes that occur as a consequence of disease and ageing or in response to treatment. It is an extremely useful advance in neuroimaging technology and one that will undoubtedly have many clinical uses in the near future. PMID- 11929437 TI - Alcoholism and personality. AB - OBJECTIVE: The search for an alcoholic personality has been pursued with varying enthusiasm throughout the 20th century. This paper reviews the methodological issues, research designs and current theories relating alcoholism and personality. METHOD: A selected literature search using computerised databases was ordered via the four major research design strategies: cross sectional studies, high-risk studies, longitudinal studies and genetic epidemiology studies. RESULTS: Cross sectional studies have suggested that two broad bands of personality, impulsivity/novelty seeking and neuroticism/negative emotionality, are associated with alcoholism. Although high-risk studies have repeatedly shown that sons of male alcoholics are at increased risk of alcoholism, whether this risk is related to personality variables is unclear. Many authors believe that the presence of antisocial personality disorder is a confounder and that this may explain some of the contradictory findings. Longitudinal studies have consistently reported that antisocial behaviour and hyperactivity are related to later alcoholism. Negative emotionality seems to be less important and may largely be a consequence of the alcoholism itself. Genetic epidemiological studies suggest that personality measures play a modest but significant role in the genetic influence of alcoholism. The strongest relationships are with conduct disorder and antisocial behaviour. The postulated alcoholic subtypes (Type I, Type II or Type A/B) based on age of onset and personality style have been challenged by recent research. The most vulnerable to alcoholism may be those with both high impulsivity/high novelty seeking and high neuroticism/negative emotionality. CONCLUSION: Antisocial behaviour and hyperactivity are the most consistent behaviours associated with alcoholism. These behaviours are not specific for alcoholism and are associated with many other psychiatric conditions. Personality variables by themselves explain only a small proportion of the risk for alcohol dependence. There is no alcoholic personality nor are there personality measures which are specific to vulnerability to later alcohol dependence. Attempting to link alcoholism with theoretical, poorly validated models of personality is premature. PMID- 11929438 TI - Notes towards a professional ethics for psychiatry. AB - OBJECTIVE: The uniqueness of psychiatry calls for a unique ethics. By identifying the features distinguishing psychiatry as medical and social practice, this article seeks to illustrate the methodology by which that ethics can be derived and to determine what kind of a framework and focus such an ethics requires. METHOD: The author is an analytically trained philosopher and employs the method of conceptual analysis. RESULTS: At least three characteristics are suggested by the features which taken together constitute psychiatry's uniqueness: an ethical framework accommodating character, a rubric for acknowledging boundary violations, and an emphasis on gender. CONCLUSIONS: The larger task of formulating the substance and details of that unique ethics is the next step. PMID- 11929439 TI - Deinstitutionalization for long-term mental illness: a 6-year evaluation. AB - OBJECTIVE: Forty-seven patients with long-term mental illness were transferred to the community following the closure of a psychiatric hospital in Sydney. This 6 year evaluation is an extension of a detailed clinical, ethnographic and economic study of the initial 2-years of community transition. METHOD: Quantitative evaluation was conducted using the Brief Psychiatric Rating Scale, Life Skills Profile, Social Behaviour Scale, Montgomery Asberg Depression Rating Scale and Quality Of Life measures. Assessments were completed prior to discharge and at two- and 6-year intervals following community transfer. Repeated measures analysis was utilized to determine changes in outcome variables over time. The residents' perception of 6-years of community living was explored in qualitative semistructured interviews. Details of accommodation, level of care, readmissions, incidents and medication were also documented. RESULTS: During the 6 years following community relocation a total of seven residents returned to hospital for long-term care, three residents died from medical causes and one resident required detention in a corrective services facility. The 36 residents who remained in the community at the 6-year follow-up no longer required intensive 24 h supervision. Living semi-independently, 23 residents resided in two to three person accommodation with either daily or weekly case manager visits. Clinically, community residents remained stable over the 6 years without significant changes in psychiatric symptoms, depression, living skills or social behaviour problems. Clinical stability was achieved with significant reduction in medication levels over the 6 years. Community-based residents continued to experience improved quality of life and reported their marked preference for living in the community. CONCLUSION: The residents maintained community tenure with significant improvement in quality of life and a reduction in medication, supported by a mental health system with adequate community resources. Issues regarding continuing rehabilitation and social integration need to be addressed. Further deinstitutionalization will require 24-h supervision for most initially and for some on a continuing basis. An ageing population will require specific age related medical and psychiatric services. PMID- 11929440 TI - Reduction of trauma symptoms following group therapy. AB - OBJECTIVE: We examined the effect of a group programme for the treatment of adults suffering the sequelae of childhood abuse and/or neglect. Symptom reduction was the indicator of whether this group programme was an effective treatment modality. METHOD: This was an experimental field study using a prepost design and General Linear Model (GLM) analyses. The group programme was conducted for adult survivors of child abuse and neglect and took place at Specialty Clinics, Central Coast Mental Health Services. There were 83 participants, comprising 64 women and 19 men; 56 were in the experimental group, and 17 in the control (waitlist) group. The Trauma Symptom Inventory (TSI) was administered to both experimental and control groups prior to receiving treatment and again three months later. RESULTS: General Linear Model analyses indicated there was a reduction of trauma symptoms for the experimental group on seven of the 10 scales of the TSI and on the three composite scales; no similar reduction was found for the control group. CONCLUSIONS: The study indicates that this group programme may have been an effective treatment modality for abuse survivors. The level of symptomatology was reduced for those in the group programme compared with the control group for whom there were no changes. PMID- 11929441 TI - The roles and relationships of psychiatrists and other service providers in mental health services. AB - OBJECTIVE: A number of the problems facing psychiatry are identified as having common origins in unresolved issues about teamwork. The aim is to identify the issues and to formulate recommendations for dealing with these obstacles. METHOD: The Royal Australian and New Zealand College of Psychiatrists Professional Liaison Committee (Australia) of the Board of Professional and Community Relations convened a number of meetings and discussions, with external assistance from JW and TT. RESULTS: Much of contemporary mental health care is delivered either directly or indirectly by several persons, often working in teams. Teamwork and collaboration are central to good working relationships and service delivery. Effective teamwork and collaboration is supported by key elements including agreed goals, an agreed approach, effective communication styles, established ground rules, clear team roles, and competent leadership. The obstacles to effective teamwork and collaboration are discussed in terms of the following: ambiguity and conflict over roles; conflict and confusion over leadership; differing understandings of clinical responsibility and accountability; interprofessional misperceptions; and differing rewards between the professions. CONCLUSIONS: Developing an understanding of these issues opens the way to a series of recommendations in relation to education, professional organizations, and workplaces and government, which can be considered by a number of sectors and groups. PMID- 11929442 TI - Suicide in Australian farming, 1988-1997. AB - OBJECTIVES: To identify and describe suicide data for occupational classifications relating to farm managers and agricultural labourers for Australia for the period 1988 to 1997, and to relate resultant suicide rates to farmers' terms of trade. METHOD: Descriptive and linear regression analysis of aggregated mortality data. RESULTS: In the period 1988 to 1997, 921 suicides were identified. The majority of these suicides were farm managers (67.4%). Farm manager suicides occurred predominantly in older age groups (55 + years). In comparison agricultural labourer suicides were younger, with the majority of suicides occurring in the 15-39 years age group. The most common methods for both groups were firearms (particularly hunting rifles and shot guns), hanging and motor vehicle exhaust gas. These methods accounted for approximately 81% of all male farm suicides. Firearms accounted for 51% of male farm suicide, in comparison to 23% for the wider Australian male population for the same period. Estimated age standardized rates per 100 000 for male farm managers for the 10 year period ranged from 24.8 to 51.4. For male agricultural labourers these rates ranged from 23.5 to 41.9. Analyses also revealed a negative correlation (r = 0.75, p < 0.01) between farm manager suicide rates and farmers' terms of trade. CONCLUSION: Male farm manager and agricultural labourer suicide rates are higher than male national rates and rates in the wider rural population, particularly in the later years of the period investigated. Firearms prefigure as the most common method of suicide, despite decreases in this method in the wider rural population. Establishing more accurate denominator data for the farming community is a priority, as is preventative action that accounts for unique aspects of farm life. PMID- 11929443 TI - Suicide among psychiatric patients: a case-control study. AB - OBJECTIVE: To examine patient- and treatment-based differences between psychiatric patients who do and do not die by suicide. METHOD: By linking databases of deaths and psychiatric service use in Victoria, we compared 597 cases who suicided over 5 years with individually matched controls. RESULTS: Cases and controls could not be distinguished on the majority of patient- or treatment-based characteristics. The exceptions were that cases were more likely to be male, less likely to be outside the labour force, more likely to have recent contact with inpatient and community services, and more likely to have a registration as their last contact. CONCLUSION: Patients who suicide 'look' similar to those who do not, suggesting prevention approaches should ensure that all psychiatric patients receive optimal care, including appropriate detection, diagnosis, assessment and treatment of mental health problems, and careful, individualised assessment of suicide risk. PMID- 11929444 TI - Clinical supervision: a review of underlying concepts and developments. AB - OBJECTIVE: This review examines how psychiatric clinical supervision is represented in the psychiatric literature and its relevance for Australasian psychiatry. METHOD: The literature was first identified then reviewed using Medline and Psychlit, manual searches of relevant journals and personal contact with some key workers in Australia and New Zealand. RESULTS: The predominantly American literature written two to three decades ago reflected the conditions in which psychiatry was practised at that time, largely based in asylums or private offices and informed by the dominant psychoanalytic discourse of that era. These articles, frequently anecdotal and with little empirical support, conceptualized supervision as a developmental process, a syndrome, or a process of identification. They focused substantially on the nature of the relationship between the trainee and supervisor. More recent writers have included trainees' perspectives. They have identified a number of problems with supervision, including role conflicts, uncertainty about boundaries, lack of supervisory training and lack of effective feedback, and have introduced the concepts of adult learning as highly relevant. These concerns, however, have led to little change to date. CONCLUSION: The implementation by the Royal Australian and New Zealand College of Psychiatrists (RANZCP) of new training by-laws provides an opportunity to define the meaning of supervision in the current clinical context, to undertake research to clarify the key elements in the process, and to evaluate different techniques of supervision. PMID- 11929445 TI - Informing patients about tardive dyskinesia: four-year follow up of a trial of patient education. AB - OBJECTIVE: This paper evaluates the effects of an educational intervention about tardive dyskinesia on knowledge and clinical stability at long-term follow up. METHOD: Fifty-six patients receiving antipsychotic maintenance completed a questionnaire assessing their knowledge about tardive dyskinesia. After random allocation to either educational intervention or control group, their knowledge, clinical stability and rates of tardive dyskinesia were reassessed after four years. RESULTS: Seventy per cent of patients completed the study. The patients in the educational group retained significantly more knowledge at follow up than at baseline but this knowledge was not significantly greater than that of the control group. There were no significant differences in the clinical outcomes between the groups. CONCLUSION: Patients can retain a small but significant amount of information with a low risk of noncompliance. Discussion about tardive dyskinesia is necessary in the process of obtaining informed consent to treatment. PMID- 11929446 TI - High prevalence disorders in urban and rural communities. AB - OBJECTIVE: High prevalence disorders (anxiety, depressive and substance use) are generally assumed to be more common in urban than rural dwellers. The aims of this paper are (i) to critically review studies measuring prevalence in rural as opposed to urban location, and (ii) to argue the need to look beyond the 'quantity' question to the quality question: how does urban or rural place influence mental health? METHOD: A literature review (Medline and PsychLIT) was carried out using the words 'rural, urban, mental/psychiatric, illness/disorders and prevalence', as well as a review of relevant papers and publications known to the authors. RESULTS: Many studies examining urban/rural differences in the rate of high prevalence disorders have been reported. Most use a 'one size fits all' definition of urban and rural, which assumes location is the key issue. The majority fail to show the purported difference in prevalence between the two settings. In general, studies have not examined interaction effects, but have simply treated the independent variables as main effects. Available data suggest that a variety of socio-demographic factors are more powerful predictors of difference in prevalence than is the location of residence. CONCLUSION: Further studies are required to understand if and how rural or urban place contributes to the development of psychiatric morbidity. These studies should mirror the clinical situation by taking into account a variety of individual and community based (including urban/rural place) risk factors which may be important determinants of mental health and mental illness, and examining the interaction between them. This may then identify the nature of any differences or what issues are specific to, or especially important, in the rural setting. PMID- 11929447 TI - The prevalence and nature of stalking in the Australian community. AB - OBJECTIVE: This study examines the extent and nature of stalking victimisation in a random community sample. METHOD: A postal survey was distributed to 3700 adult men and women selected from the electoral roll in the State of Victoria. Outcome measures included the lifetime and annual cumulative incidence of stalking, the duration and methods of harassment, rates of associated violence and responses to victimisation. RESULTS: Almost one in four respondents (23.4%;432) had been stalked, the unwanted behaviour they were subjected to being both repeated and fear-provoking. One in 10 (197) had experienced a protracted course of stalking involving multiple intrusions spanning a period of at least one month. Women were twice as likely as men to report having been stalked at some time in their lives, though the rates of victimisation in the 12 months prior to the study did not differ significantly according to gender. Younger people were significantly more likely than older respondents to report having been stalked. Victims were pursued by strangers in 42% of cases. The most common methods of harassment involved unwanted telephone calls, intrusive approaches and following. Associated threats (29%) and physical assaults (18%) frequently arose out of the stalking. Significant social and economic disruption was created by the stalking for 63% of victims. Most sought assistance to manage their predicament (69%). CONCLUSIONS: The experience of being stalked is common and appears to be increasing. Ten percent of people have been subjected at some time to an episode of protracted harassment. Assaults by stalkers are disturbingly frequent. Most victims report significant disruption to their daily functioning irrespective of exposure to associated violence. PMID- 11929448 TI - Factors associated with dysmorphic concern and psychiatric morbidity in plastic surgery outpatients. AB - OBJECTIVES: To compare new referrals to a plastic surgery clinic for cosmetic (non-medically explained) reasons with a control group of equal size with medically explained symptoms. METHOD: Patients attending for cosmetic (non medically explained) reasons were compared with the controls using the general health questionnaire (GHQ), and dysmorphic concern questionnaire (DCQ). Patients were divided into high and low DCQ scores on the basis of their median scores. RESULTS: Ninety subjects were approached of whom 84 (93%) participated giving 42 patients each in the cosmetic (non-medically explained) and control groups. Forty four per cent were referred for mammoplasty (n = 37) and 8% for rhinoplasty (n = 7). The other 40 cases (48%) were for other procedures including excision, abdominoplasty and blepharoplasty. Thirty-two per cent of the sample were GHQ cases (n = 27). Patients presenting for cosmetic (non-medically explained) reasons were 13 times more likely to be female (95% CI = 4.3-41), nine times more likely to have high DCQ scores (95% CI = 3.3-24), six times more likely to be GHQ cases (95% CI = 2.1-17), and seven times more likely to present for mammoplasty. The same factors were associated with high DCQ scores. Patients with high DCQ scores were 32 times as likely to be GHQ cases (95% CI = 6.8-151). On multivariate analysis, dysmorphic concern emerged as the only independent predictor of GHQ caseness rather than sex, surgical diagnosis or procedure (adjusted OR = 32.0, 95 % CI = 6.5-156). Similarly, only GHQ caseness and presentation for cosmetic (non-medically explained) surgery independently predicted DCQ score. CONCLUSIONS: Patients presenting for cosmetic (non-medically explained) surgery have high rates of dysmorphic concern and psychiatric morbidity PMID- 11929450 TI - An analysis of systems of classifying mild cognitive impairment in older people. AB - OBJECTIVE: Over the past two decades, a number of systems have been developed for the classification of cognitive and behavioural abnormalities in older people, in order that individuals at high risk of developing neurodegenerative disease, particularly Alzheimer's disease, may be identified well before the disease manifests clinically. This article critically examines the inclusion and exclusion criteria of a number of such classification systems, to determine the effect that variations in criterion may have on clinical, behavioural and neuroimaging outcomes reported from older people with mild cognitive impairment. METHOD: Qualitative review of the literature describing systems of classifying mild cognitive impairment, and outcomes from clinical, behavioural, neuroimaging and genetic studies of older people with mild cognitive impairment. RESULTS: The exclusion and inclusion criteria for these classification systems vary markedly, as do the design of studies upon which the validity of these systems has been assessed. Minor changes to individual exclusion/inclusion criterion may result in substantial changes to estimates of the prevalence and clinical outcome of mild cognitive impairment, while inadequate experimental design may act to confound the interpretation of results. CONCLUSIONS: As a result of these factors, accurate and consistent estimates of the outcome of mild cognitive impairments in otherwise healthy older people are yet to be obtained. On the basis of this analysis of the literature, optimal criteria via which accurate classifications of mild cognitive impairment can be made in future are proposed. PMID- 11929449 TI - The assessment of disability in patients with psychotic disorders: an application of the ICIDH-2. AB - OBJECTIVES: To describe the application of the International Classification of Functioning and Disability (ICIDH-2) in the assessment of disability in patients with psychotic disorders. METHOD: The study was conducted at the inpatient unit of St Vincent's Mental Health Service, Melbourne. Twenty inpatients with psychotic disorders were concurrently assessed by two raters. An evaluation of the feasibility of the instrument was made during this process. Inter-rater agreement was determined using raw agreement percentages and weighted kappas. RESULTS: The ICIDH-2 provides a comprehensive framework for the description and measurement of disability, including the dimensions of impairments in body structures and functions, activity limitations, participation restrictions and environmental factors. Certain codes are difficult for clinicians to rate because of the subjective manner in which they are defined. Our inter-rater reliability assessment varied across the dimensions from poor to almost perfect agreement. CONCLUSIONS: The ICIDH-2 is potentially useful in the clinical assessment of disability in patients with psychotic disorders although modifications are needed. These could include the use of anchor points defined for each dimension or code to assist the rating process and an alternative rating scale in which categories for rating are more broadly defined. Formal training may also be necessary to enable standardization of the rating process. There is also a need to augment measurements by clinicians with the assessment of the subjective experience of patients using self-rated disability and assessment of quality of life. PMID- 11929451 TI - The mental health of young people in Australia: report by the National Mental Health Strategy. PMID- 11929453 TI - Transsexualism - need it always be a DSM-IV disorder. PMID- 11929455 TI - Connection between lithium and muscular incoordination. PMID- 11929456 TI - Clozapine associated pericarditis and elevated troponin I. PMID- 11929457 TI - Risperidone-induced Pisa syndrome. PMID- 11929458 TI - A misleading and pejorative book. PMID- 11929469 TI - Urological problems in pregnancy. AB - During pregnancy the urinary tract undergoes extensive anatomical and physiological changes. These changes can result in many symptoms and pathological conditions that may affect the mother and fetus. It is well documented that childbirth may result in urinary tract damage which may predispose to postpartum symptoms. This review describes the physiological and pathological consequences of pregnancy and delivery on the urinary tract, and how these may be minimized. PMID- 11929470 TI - Advances in uroradiological imaging. AB - The development of new imaging techniques and the refinement of established methods in uroradiological imaging is proceeding rapidly. In the last few years several important developments have been implemented in the routine diagnostic evaluation of urological patients.A milestone is the recent advent of multidetector helical computed tomography (CT), enabling the radiologist to provide the clinician with high-quality three-dimensional (3-D) reconstructions of the urological organs. Powerful workstations are an indispensable tool in the post-processing of CT and magnetic resonance imaging (MRI)data. Significant advances in imaging were obtained in the fields of oncological imaging (e.g. prostate MRI and spectroscopic imaging), paediatric uroradiology(e.g. MR urography) and the evaluation of stone disease by unenhanced helical CT. PMID- 11929471 TI - Principles of nephrectomy for malignant disease. AB - For many years the prevailing belief was to advocate'radical' nephrectomy via a transperitoneal approach as the standard surgical procedure for renal cell carcinoma (RCC). because the early control of the renal vessels before manipulating the kidney should minimize the likelihood of disseminating tumour cells during surgery. This philosophy was based on retrospective data which were never confirmed in a controlled trial. Since then,evidence has accumulated that some patients maybe better served by an extraperitoneal (translumbar)approach, providing similar oncological efficacy with the added advantage of reduced morbidity. However,these results are again either retrospective or statistically insignificant, and therefore do not allow firm conclusions. Nevertheless, if there is any difference in the possible intraoperative dissemination of tumour, depending on the type of surgical approach, it will be small, requiring analysis in a large randomized multicentre trial. The treatment of choice for disease that is not disseminated is surgery, although the 5-year survival rates for all stages do not exceed 60%, even in contemporary series. Further improvements will probably have to rely on the development of more effective systemic therapy and the application of combined treatments to counter the relatively many patients presenting with advanced stages. Concepts and progress in this field appear to be of major interest for modern uro-oncologists after the advent of immunotherapeutic strategies that require a surgical intervention at some stage of the treatment cascade. PMID- 11929472 TI - Interdisciplinary abdomino-urological surgery for advanced colorectal carcinoma involving the urogenital tract. PMID- 11929473 TI - Breastfeeding and the development of atopic disease during childhood. PMID- 11929474 TI - The allergen challenge. PMID- 11929475 TI - Matrix metalloproteinase-9: marker or mediator of tissue damage in asthma? PMID- 11929476 TI - Changing views on carbon monoxide. PMID- 11929477 TI - Transforming growth factor-beta1: its anti-inflammatory and pro-fibrotic effects. PMID- 11929478 TI - Considerations on third generation antihistamines. PMID- 11929479 TI - Increased incidence of skin infections in atopy: evidence for an antigen-specific homing defect? PMID- 11929480 TI - Alcoholic drinks and asthma. PMID- 11929481 TI - Side-chain-specific reactions to betalactams: 14 years later. PMID- 11929482 TI - Isocyanate-induced occupational asthma in far-east Asia: pathogenesis to prognosis. PMID- 11929483 TI - Breastfeeding duration is a risk factor for atopic eczema. AB - BACKGROUND: The results of numerous studies on the influence of breastfeeding in the prevention of atopic disorders are often contradictory. One of the most important problems is confounding by other lifestyle factors. OBJECTIVE: The aim of the present study was to analyse the effect of any breastfeeding duration on the prevalence of atopic eczema in the first seven years of life taking into account other risk factors. METHODS: In an observational birth cohort study 1314 infants born in 1990 were followed-up for seven years. At 3, 6, 12, 18, 24 months and every year thereafter, parents were interviewed and filled in questionnaires, children were examined and blood was taken for in vitro allergy tests. Generalized Estimation Equations (GEE)-models were used to model risk factors for the prevalence of atopic eczema and for confounder adjustment RESULTS: Breastfeeding was carried out for longer if at least one parent had eczema, the mother was older, did not smoke in pregnancy, and the family had a high social status. The prevalence of atopic eczema in the first seven years increased with each year of age (OR 1.05; 95% CI 1.01-1.09 for each year), with each additional month of breastfeeding (1.03; 1.00-1.06 for each additional month), with a history of parental atopic eczema (2.06; 1.38-3.08), and if other atopic signs and symptoms appeared, especially specific sensitization (1.53; 1.25-1.88), and asthma (1.41; 1.07-1.85). Although breastfeeding should be recommended for all infants, it does not prevent eczema in children with a genetic risk. CONCLUSION: Parental eczema is the major risk factor for eczema. But in this study, each month of breastfeeding also increased the risk. PMID- 11929484 TI - Removal of bronchoalveolar cells augments the late eosinophilic response to segmental allergen challenge. AB - BACKGROUND: In patients with quiescent asthma, macrophages are the most prevalent cells recovered by bronchoalveolar lavage (BAL). Through activation via their FcepsilonRII receptors or by acting as antigen-presenting cells, macrophages could, in theory, promote the late airway response to allergen. OBJECTIVE: In order to investigate the importance of macrophages and other airway luminal cells in inducing the late airway response, a novel washout experiment was designed. METHODS: Five patients with ragweed-allergic asthma underwent bronchoscopy and segmental bronchial challenge with either normal saline or short ragweed extract in two segments of one lung. In a third segment of the opposite lung, 12 successive BALs (25 mL each) were performed, followed by challenge with an identical dose of short ragweed (washed-challenged segment). After 24 h, all three challenged segments underwent BAL. RESULTS: Initially, in the washed challenged segment, over 80% (mean 80.4%, range 68-88%) of the recoverable airway dwelling cells were removed. Unexpectedly, 24 h later these same washed challenged segments contained more eosinophils in the BAL than the challenged segments from the opposite lung (P = 0.033). CONCLUSIONS: Removing the majority of airway luminal cells followed by allergen bronchoprovocation increased the number of eosinophils recovered 24 h after challenge. Our results suggest that in quiescent allergic asthma, the airway luminal cells are protective and attenuate the late eosinophilic response to allergen challenge. PMID- 11929486 TI - Increased carbon monoxide levels in the nasal airways of subjects with a history of seasonal allergic rhinitis and in patients with upper respiratory tract infection. AB - BACKGROUND: Carbon monoxide (CO) has emerged as an endogenously produced gaseous mediator known to be involved in bronchial smooth muscle regulation. Increased amounts of CO have been found in exhaled air during asthma and lower airway inflammation. Recently CO has been shown to be produced in the nasal airways, but there are no reports of altered CO levels in nasal airways during inflammation. OBJECTIVE: This study was designed to investigate if CO levels increase in the human nasal airways during inflammatory conditions, such as allergy and upper airway respiratory tract infection (URTI). METHODS: CO was sampled separately from the upper and lower airways of 13 healthy control subjects, six patients with a history of allergic rhinitis and six patients with URTI. RESULTS: Nasal CO levels were increased in subjects with allergic rhinitis, compared to healthy controls (2.07 +/- 0.15 ppm, n = 6 and 1.62 +/- 0.08 ppm, n = 13, respectively, P < 0.01). CO levels were also increased in patients with URTI, compared to the same controls (1.92 +/- 0.09 ppm, n = 6, P < 0.05). Normal levels of CO were found in air from the lower airways among subjects with allergic rhinitis, whereas corresponding levels in the URTI patients were increased. CONCLUSION: The present data demonstrates that upper airway CO levels increase in parallel with different inflammatory stimuli, such as allergy and infection, suggesting a role for CO as marker or mediator of nasal inflammation. PMID- 11929485 TI - Increased release of matrix metalloproteinase-9 in the plasma of acute severe asthmatic patients. AB - BACKGROUND: Matrix metalloproteinases (MMPs) are likely to be relevant mediators of the extracellular matrix (ECM) degradation and airway remodelling. OBJECTIVE: We have compared the levels of MMPs, eotaxin and soluble interleukin 2 receptor (IL-2R) in the plasma of healthy subjects, atopic patients and asthmatic patients. METHODS: The asthmatic patients were separated into two groups, either well controlled on inhaled therapy or acute severe asthma. Patients with acute severe disease had all received systemic corticosteroids from 12 to 48 h before the blood was taken. Blood was recovered in EDTA tubes, incubated with either f MLP, PMA or vehicle for 10 min and centrifuged. MMP-9, TIMP-1, IL-2R and eotaxin levels were measured in the plasma by ELISA. Moreover, the activity of MMPs was also evaluated by zymography. RESULTS: An increased basal level of MMP-9 and IL2 R was observed in acute severe asthma. Following stimulation with f MLP and PMA there was an enhanced production of MMP-9 in the plasma of all groups of patients. However, the MMP-9 level was significantly enhanced in acute severe asthma, compared with the others. No difference was found for the TIMP-1 level between the patients. The eotaxin level in plasma was found to be significantly lower in acute severe asthmatics compared with the others groups. Zymography technique showed a significant increased activity of MMP-9 (92 kDa) but not MMP-2 (66 kDa) in the plasma of patients with acute asthma. CONCLUSION: The increased in MMP-9 production and activity observed in the present study suggests a process of extracellular matrix degradation in acute severe asthmatic patients and proposes MMP-9 as a non-invasive systemic marker of inflammation and airway remodelling in asthma. PMID- 11929487 TI - Effects of beta-endorphin on nasal allergic inflammation. AB - BACKGROUND: Beta-endorphin is a derivative of pro-opiomelanocortin. Cells of the immune system can also synthesize and secrete beta-endorphin. Its concentration is increased during the allergic reaction and during stress. Increased reactivity during psychological stress of allergic subjects is also well known. OBJECTIVE: Is beta-endorphin one physiological link between stress and an exacerbation of the allergic reaction? METHODS: First, intranasal beta-endorphin challenges with subsequent lavages to determine histamine and albumin levels and measurements of nasal flow and resistance in dose-response and time course experiments were performed. Secondly, we examined whether beta-endorphin pre-treatment increased the antigen-induced release of histamine and albumin in nasal lavages and the clinical symptoms. RESULTS: Exogenous beta-endorphin (100 pM-10 microM/mL) induced a dose-dependent increase in nasal symptoms in asymptomatic allergic subjects with rhinitis (n = 14) as well as in non-allergic controls (n = 10), but did not release any mediators into nasal secretion. However, comparing the antigen-evoked release of mediators into nasal secretions with that of a beta endorphin pre-treated antigen challenge we could note a significant enhancement of human serum albumin influx (P < 0.05) and histamine liberation (P < 0.05) 10 min after antigen challenge compared with the allergen challenge alone, with also a correlation with the more pronounced decrease in nasal flow (P < 0.05). CONCLUSION: These results suggest that beta-endorphin-induced increase in nasal congestion is mediated through direct neuroendocrine receptor activation independent of mast cell activation and that during the allergic reaction there is a beta-endorphin/mast cell interaction that enhances the mediator response to nasal allergen challenge. PMID- 11929488 TI - Human mast cells stimulate fibroblast proliferation, collagen synthesis and lattice contraction: a direct role for mast cells in skin fibrosis. AB - BACKGROUND: Mast cells, the key cells of immediate hypersensitivity type reactions, have also been postulated to have a central role in influencing tissue remodelling and fibrosis occurring in the skin. OBJECTIVE: Our aim was to investigate the direct role of human mast cells (HMC) in skin fibrotic processes, by assessing the effects of the addition of the human mast cell line HMC-1 to human skin fibroblasts, and to identify the responsible mediators. METHODS: HMC-1 sonicates were added to human skin fibroblasts and the following parameters were evaluated: proliferation ([3H]-thymidine), collagen synthesis ([3H] proline), activity of matrix metalloproteinases (MMPs) (zymography) and tissue inhibitors of metalloproteinases (TIMPs) (reverse zymography), and collagen gel contraction. RESULTS: HMC-1 sonicate increased significantly both proliferation and collagen production in the human skin fibroblasts and these properties were not affected by heating of the sonicate (56 degrees C, 30 min, or 100 degrees C, 3 min). Two main mast cell mediators, histamine and tryptase, were found to be responsible for the increase in fibroblast proliferation and collagen production. HMC-1 sonicate did not display any pre-formed gelatinase activity, and its addition to the fibroblasts did not change their pro-MMP-2 and MMP-2 activity. On the other hand, HMC-1 were found to possess TIMP-1 and TIMP-2. Addition of HMC-1 had no effect on fibroblasts TIMP-1 but induced a dose-dependent increase of TIMP-2 activity. In addition, HMC-1 sonicate seeded together with the fibroblasts in tri dimensional collagen gel significantly enhanced their contraction. CONCLUSION: We have shown that human mast cells, by granule-stored and therefore quickly releasable mediators, increase human skin fibroblast proliferation, collagen synthesis, TIMP-2 and collagen gel contraction. Therefore, mast cells have a direct and potentiating role in skin remodelling and fibrosis. PMID- 11929489 TI - Low prevalence of atopy in young Danish farmers and farming students born and raised on a farm. AB - BACKGROUND: Recent studies have shown that in several countries atopic sensitization to common allergens (common atopy) and atopic symptoms are markedly less prevalent in children living on a farm, compared with non-farm children living in the same rural areas. Living conditions on farms may, however, vary largely between different countries. It is also not yet known whether the "protective" effect of a farm environment can also be found in adults. MATERIALS AND METHODS: Common atopy and respiratory health were assessed by skin prick tests (SPT), questionnaire and measurement of bronchial hyper-responsiveness (BHR) in the Sund Stald (SUS) study, a cohort study on respiratory health in Danish farming students and conscripts from the same rural areas as controls. Results of SPT were confirmed by IgE serology in all SPT+ subjects and a subset of SPT- subjects. Prevalences of common atopy, respiratory symptoms and bronchial hyper- responsiveness were compared for farmers and controls, and for those who had or had not lived on a farm in early childhood. RESULTS: In multiple logistic regression analyses adjusting for ever smoking and a familial history of allergy, both being a farmer (ORs 0.62-0.75) and having had a farm childhood (ORs 0.55 0.75) appeared to contribute independently to a lower risk of sensitization to common allergens as assessed by SPT and IgE serology. A farm childhood was also inversely associated with high total IgE (OR 0.68), presence of respiratory symptoms (ORs 0.69-0.79) and BHR (OR 0.61) in these analyses. Direction and strength of the association between being a farmer and respiratory symptoms or BHR varied widely (ORs 0.69-1.28). CONCLUSION: The "anti-atopy" protective effect of a farm childhood could be confirmed in Danish farming students: prevalences of positive SPT, specific and total IgE, allergic symptoms and BHR were lower in those being born or raised on a farm. Past exposure to the farm environment in early childhood may therefore also contribute to a lower risk of atopic sensitization and disease at a later age. PMID- 11929490 TI - A lower prevalence of atopy symptoms in children with type 1 diabetes mellitus. AB - BACKGROUND: The Th1/Th 2 concept is a model to understand the pathophysiology of certain diseases. Atopic diseases (asthma, eczema and hayfever) are characterized by a chronic inflammatory reaction that is dominated by Th 2 cells, and type 1 diabetes mellitus (DM) is Th1 cell dominated. Because it is known that Th1 and Th 2 cells reciprocally counteract each other, it can be speculated that the prevalence of Th 2-mediated disease is lower in patients with Th1-mediated disease. OBJECTIVE: To compare the prevalence of atopic diseases between children with DM and age-matched controls. METHODS: Parents of children with DM were requested by Dutch paediatricians to complete the International Study of Asthma and Allergies in Childhood (ISAAC) questionnaire on the prevalence of atopic diseases. A control group was derived from a Dutch cross-sectional survey (the ISAAC2 study). RESULTS: We received 555 completed questionnaires, which is estimated to be 25% of the total number of Dutch children with DM. The control group consisted of 777 children. After age-matching, the questionnaires of 188 DM patients were used. Symptoms of asthma, hayfever and eczema were reported less in the group of children with DM compared with the control group (wheeze last year, OR 0.796, 95% CI 0.408-1.554; hayfever symptoms last year, OR 0.642, 95% CI 0.369 1.118; eczema symptoms last year, OR 0.693, 95% CI 0.430-1.115). CONCLUSION: The lower prevalence of astma, hayfever and eczema symptoms in DM patients compared with age-matched controls, although not statistically significant, is consistent with the Th1/Th 2 concept. PMID- 11929491 TI - Stressful life events promote the manifestation of asthma and atopic diseases. AB - BACKGROUND: Psychosocial stress is known to aggravate asthma. Less is known about the impact of stressful life events on the expression of asthma and atopic disorders. OBJECTIVE: To determine whether the onset of asthma, allergic rhinitis or conjunctivitis, and atopic dermatitis, are associated with stressful life events. METHODS: A postal survey on risk factors for asthma and atopic diseases was carried out among 10 667 Finnish first-year university students aged 18-25 years. Stressful life events, (i) severe disease or death of a family member, and (ii) parental or personal conflicts, were retrospectively recorded during the preceding year, 1-5 years, 6-10 years, and more than 10 years prior to the survey response. In a case-control setting, conditional multiple logistic regression analysis was used to assess the temporal association between major stressful life events occurring during a period either preceding, concomitant or subsequent with subject's diagnoses. RESULTS: Concomitant parental and personal conflicts increased the risk of asthma (OR 1.72, 95% CI 1.10-2.69) when adjusted by parental asthma, education and passive smoking at early age. Concomitant severe disease or death of mother, father or spouse (OR 1.52, 95% CI 1.09-2.16) and precedent parental and personal conflicts (OR 1.75, 95% CI 1.15-2.77) increased the risk of manifestation of allergic rhinoconjunctivitis when adjusted for parental atopic disease, education and passive smoking. Subjects' asthma and atopic dermatitis, but not allergic rhinoconjunctivitis, were related to excess of subsequent stressful life events. CONCLUSION: An association between stressful life events and subjects' asthma, allergic rhinoconjunctivitis and atopic dermatitis is suggested. PMID- 11929492 TI - A randomized, double-blind trial of the effect of glucocorticoid, antileukotriene and beta-agonist treatment on IL-10 serum levels in children with asthma. AB - BACKGROUND: Levels of an immunoregulatory and anti-inflammatory cytokine IL-10 are reduced in asthmatic airways, potentially contributing to more intense inflammation. Triamcinolone has anti-inflammatory properties and the anti inflammatory effects of montelukast and formoterol have been discussed. OBJECTIVE: The purpose of this study was to define the effect of treatment with triamcinolone, montelukast and formoterol on the serum level of IL-10, eosinophil blood counts, eosinophil cationic response (ECP) and clinical parameters (symptom score, FEV1 and PC20H) in children with moderate asthma. METHODS: An 8-week, placebo-controlled and randomized, double-blind trial was carried out. The subjects were 91 children with moderate atopic asthma who were allergic to dust mite. Patients were randomly allocated to receive 400 microg triamcinolone (n = 19), 5 or 10 mg (according to age) montelukast (n = 18), 24 microg formoterol (n = 18) or placebo (n = 36). RESULTS: Seventy-nine children completed the study. After treatment with triamcinolone and montelukast the level of IL-10 in blood serum significantly increased, eosinophil blood counts and ECP levels significantly decreased and all clinical parameters improved; treatment with formoterol had no effect on IL-10 level, eosinophil blood counts in serum and bronchial hyper-reactivity; ECP level significantly decreased after treatment and asthma symptoms and FEV1 improved significantly. Mean IL-10 levels in serum before and after treatment with triamcinolone were 7.23 pg/mL with 95% CI, 6.74 7.72% and 14.24 pg/mL with 95% CI, 11.6-16.88%, respectively (P < 0.001); with montelukast they were 6.59 pg/mL with 95% CI, 6.26-7.23% and 10.94 pg/mL with 95% CI, 8.24-12.65%, respectively (P < 0.002); with formoterol they were 7.06 pg/mL with 95% CI, 6.61-7.52% and 7.04 pg/mL with 95% CI, 6.15-7.93%. We found statistically significant correlations between serum level of IL-10 and serum level of ECP after treatment with triamcinolone and montelukast. CONCLUSION: This study demonstrates that one possible way by which triamcinolone and montelukast contribute to inhibition of inflammation is by increasing IL-10 levels. PMID- 11929493 TI - Controlled administration of penicillin to patients with a positive history but negative skin and specific serum IgE tests. AB - BACKGROUND: Although subjects with a positive history of immediate allergy to penicillin and negative skin test are traditionally considered to tolerate penicillin, current evidence indicates that they may develop an immediate reaction despite negative skin and serum specific IgE tests. It is thought that these patients require additional tests to confirm the diagnosis. OBJECTIVE: To assess in a large group of patients with a history of immediate allergy to penicillins but with both skin test and CAP-FEIA-negative to classical and side chain penicillin determinants, the role of controlled administration of betalactams as a diagnostic test. METHODS: A group of 330 patients with a history of immediate allergic reactions to penicillins was studied by two evaluators from the same allergy unit using the following protocol: skin tests with major and minor determinants of benzylpenicillin (benzylpenicilloyl-poly l-lysine and minor determinant mixture), amoxicillin and ampicillin, and determination of specific IgE antibodies to penicillins, by CAP-FEIA, in serum. If both tests proved negative, a controlled administration of the drug was then carried out. RESULTS: A total of 89 (27%) patients were skin test and CAP-FEIA-negative and therefore required controlled administration of the drug. Of these, 49 developed an immediate response and were therefore considered allergic, and the remainder had good tolerance after administration of both benzylpenicillin and amoxicillin. The clinical characteristics of this group were similar to the other allergic patients who were skin test or CAP-FEIA-positive, except that they were younger (P < 0.01). Twenty-two (45%) developed a response to benzylpenicillin and 27 (55%) had a selective response to amoxicillin. Although all reactions appeared within 1 h, a positive correlation was found between the dose inducing the response and the time elapsed from drug administration, for both benzylpenicillin and amoxicillin (P < 0.001). CONCLUSION: These data indicate that an important number of subjects are not correctly identified if only skin tests and/or CAP FEIA are used and that this is particularly relevant for side chain-specific reactions and younger subjects. This suggests that new diagnostic tests are required so as to limit the use of controlled administration. PMID- 11929494 TI - Flow cytometric basophil activation test by detection of CD63 expression in patients with immediate-type reactions to betalactam antibiotics. AB - BACKGROUND: In this study, we used flow cytometry to determine the percentage of activated basophils that expressed the CD63 marker after in vitro stimulation by different betalactam antibiotics. The diagnostic reliability of the technique was assessed, as well as its correlation with specific IgE. METHODS: Fifty-eight patients with clinical allergy to betalactam antibiotics and presenting positive skin tests to at least one of the allergens (minor determinant mixture (MDM), benzylpenicilloyl-polylysine (PPL), penicillin, ampicillin, amoxicillin, cephalosporins) were tested. Thirty subjects non-allergic to betalactams were also studied as controls. The flow assay stimulation test (FAST) uses flow cytometry to determine the percentage of basophils that express CD63 as an activation marker after in vitro stimulation with allergen. Double labelling with monoclonal antibodies anti-CD63-PE and anti-IgE FITC was used. RESULTS: The allergic patients show a statistically greater number of activated basophils than the control subjects, after the incubation of cells with all the betalactams at various concentrations. The sensitivity of the technique is 50%, the specificity 93.3%, the likelihood ratio for a positive value 7.46 and the likelihood ratio for a negative value 0.54. In spite of having a greater sensitivity (37.9%) and specificity (86.7%) than CAP, differences between sensitivity and specificities of both techniques (CAP and FAST) do not reach statistical significance. CONCLUSION: The basophil activation test is a particularly useful technique in the diagnosis of patients with IgE-mediated allergy to betalactams and allows the identification of 50% of patients. Used in conjunction with CAP, it allows the identification of 65.5% of such patients. PMID- 11929495 TI - Effect of formaldehyde on the expression of adhesion molecules in nasal microvascular endothelial cells: the role of formaldehyde in the pathogenesis of sick building syndrome. AB - BACKGROUND: Formaldehyde is associated with sick building syndrome (SBS), a set of diffuse and irritative symptoms predominantly involving the eyes and the respiratory tract. However, its pathophysiological mechanism in SBS has not yet been clarified. OBJECTIVE: In this study we investigated the effect of formaldehyde on the expression of adhesion molecules on human mucosal microvascular endothelial cells (HMMECs). Furthermore, we investigated the effect of formaldehyde on adhesiveness of HMMECs to eosinophils. MATERIALS AND METHODS: HMMECs were incubated with various concentrations of formaldehyde (1 ng/mL-1 microg/mL) for 24 h, and the expressions of intercellular adhesion molecule-1 (ICAM-1) and vascular cell adhesion molecule (VCAM-1) on HMMECs were evaluated by flow cytometry. The change in the expression of ICAM-1 and VCAM-1 mRNA was then evaluated by reverse transcriptase polymerase chain reaction. To understand the role of formaldehyde in eosinophilic inflammation of the nasal mucosa, we examined the effects of formaldehyde on the adhesiveness between HMMECs and eosinophils by eosinophil adhesion assay. RESULTS: Formaldehyde increased the surface expressions of ICAM-1 and VCAM-1 on HMMECs. Formaldehyde also induced ICAM-1 and VCAM-1 mRNA. In addition, the adhesiveness between HMMECs and eosinophils was also increased by formaldehyde. CONCLUSION: These in vitro studies suggest that formaldehyde may play a role as the irritant of the nasal mucosa by increasing the expressions of adhesion molecules on HMMECs and by enhancing the adhesiveness between HMMECs and eosinophils. PMID- 11929496 TI - Increased HLA-DR expression after exposure of human monocytic cells to air particulates. AB - BACKGROUND: The expression of HLA-DR on the cell membrane of antigen-presenting cells is of major importance for the induction of an allergic response in the airways. Environmental particulates are thought to play an important role in inducing or enhancing allergic sensitization, possibly by increasing the expression of HLA-DR on the cell membrane of antigen-presenting cells. In addition, these particulates may synergize with common sensitizing agents in inducing or enhancing HLA-DR and thus antigen presentation. OBJECTIVE: In this study, we investigated the potential of three particle types, namely carbon black, diesel exhaust particles and urban air particulates (0.1-1000 ng/cm(2)), to induce the expression of HLA-DR on differentiated THP-1 cells, taken as a model for alveolar macrophages. We also assessed the "adjuvant" potential of the particles on interferon (IFN)-gamma, a known enhancer of HLA-DR. RESULTS: By themselves, the particles (0.1-1000 ng/cm(2)) were not able to induce HLA-DR on the THP-1 cells after an incubation of 48 h. However, even at very low concentrations, carbon black (from 1 ng/cm(2) on) and diesel exhaust particles (from 0.1 ng/cm(2) on), interacted with IFN-gamma (100 U/mL) to enhance HLA-DR expression (up to 2.5-fold increase). CONCLUSION: This finding may reflect in vitro one of the mechanisms by which pollutant particles exert an "adjuvant" activity and may partially explain how exposure to particles can be related to the enhancement of allergic sensitization. PMID- 11929497 TI - DNA microarrays to study gene expression in allergic airways. AB - BACKGROUND: Allergic rhinitis results from interactions between a large number of cells and mediators in different compartments of the body. DNA microarrays allow simultaneous measurement of expression of thousands of genes in the same tissue sample. OBJECTIVE: To study gene expression in nasal mucosal biopsies from patients with allergic rhinitis using DNA micro-arrays. METHODS: Nasal biopsies were obtained from 14 patients with symptomatic birch pollen-induced allergic rhinitis and five healthy controls. RNA was extracted from the biopsies and pooled into one patient pool and one control pool. These were analysed in duplicate with DNA micro-arrays containing more than 12 000 known genes. RESULTS: Approximately half of the genes were expressed in the patient and control samples. Guided by the current literature we chose 32 genes of possible relevance to allergic airway inflammation and investigated their relative expression. Among these, transcripts encoding immunoglobulins and their receptors were most abundant. The expression of cytokines and growth factors was low, whereas their corresponding receptors and cell surface markers displayed higher expression levels. IgA had the highest expression of all 12 626 genes. RT-PCR showed that IgA1 was the predominant subclass. This was confirmed by the protein level in nasal fluids. Allergen-specific IgA was significantly higher in patients than in controls and correlated significantly with eosinophil granulae proteins. CONCLUSION: DNA micro-array analysis can be used to identify genes of possible relevance to allergic airway inflammation. In this study, the expression profile in the nasal mucosa was quantitatively dominated by immunoglobulins, particularly IgA. Protein analyses in nasal fluids indicated a role for allergen-specific IgA in eosinophil degranulation. PMID- 11929498 TI - Transforming growth factor-beta1 suppresses atopic dermatitis-like skin lesions in NC/Nga mice. AB - BACKGROUND: Atopic dermatitis is a chronic, relapsing inflammatory disorder characterized by pruritic and eczematous skin lesions. Transforming growth factor (TGF)-beta1 has been implicated in the suppression of inflammatory responses. OBJECTIVE: The purpose of this study is to determine whether TGF-beta1 suppresses skin lesions in a mouse model of atopic dermatitis. METHODS: We used the NC/Nga strain of mice as an in vivo model of atopic dermatitis. The effects of exogenous TGF-beta1 on atopic dermatitis-like skin lesions in NC/Nga mice were evaluated clinically, histologically and immunologically. RESULTS: Subcutaneous injection of recombinant TGF-beta1 macroscopically suppressed eczematous skin lesions in NC/Nga mice associated with reduced serum immunoglobulin E (IgE) levels. Histological analysis showed that TGF-beta1 significantly inhibited the infiltration of inflammatory cells such as mast cells and eosinophils into the skin of NC/Nga mice. Spontaneous interferon (IFN)-gamma production from splenocytes of NC/Nga mice was down-regulated by the treatment with TGF-beta1 and neutralizing antibody against IFN-gamma inhibited skin lesions in NC/Nga mice. The inhibitory effect of TGF-beta1 on the skin lesions lasted at least 1 week after cessation of the treatment. CONCLUSION: These findings indicate that TGF beta1 suppressed atopic dermatitis-like skin lesions in NC/Nga mice at least in part through down-regulation of IFN-gamma. These results suggest that TGF-beta1 may have a therapeutic potential for atopic dermatitis. PMID- 11929499 TI - Transforming growth factor-beta1 mediates coexpression of the integrin subunit alphaE and the chymase mouse mast cell protease-1 during the early differentiation of bone marrow-derived mucosal mast cell homologues. AB - BACKGROUND: Mucosal mast cells (MMC) play a central role in gut hypersensitivities and inflammation. They are morphologically, biochemically and functionally distinct from their connective tissue counterparts. Massive hyperplasia of MMC occurs 7-10 days after intestinal infection with nematodes but it has never been possible to replicate this phenomenon in vitro. OBJECTIVE: (1) To determine whether mouse bone marrow-derived mast cells (mBMMC) grown in the presence of transforming growth factor (TGF)-beta1 could develop over the same time frame (7-10 days) as MMC in parasitized mice. (2) To compare the early expression of surface receptors (integrins alphaE and beta7, c-kit and FcepsilonR) with that of the MMC-specific granule chymase mouse mast cell protease-1 (mMCP-1). METHODS: Mouse bone marrow cells were cultured in the presence of IL-9, IL-3 and Stem Cell Factor (SCF) with or without TGF-beta1. mBMMC were quantified after toluidine blue or Leishmans' staining. Expression of MMC-specific mouse mast cell proteases was analysed by ELISA, immunohistochemistry and RT-PCR. Surface antigen expression was characterized by flow cytometry and confocal microscopy. RESULTS: TGF-beta1 promotes the development of abundant MMC-like mBMMC from bone marrow progenitor cells with kinetics, which closely parallel that seen in vivo. mRNA transcripts encoding mMCP-1 and -2 are readily detectable by day 4 ex vivo in cultures grown in the presence of TGF-beta1. Between 30 and 40% and 75-90% of the cells in these cultures on days 4 and 7, respectively, have typical mast cell morphology, are c kit+, FcepsilonR+, integrin alphaEbeta7+, and express and secrete abundant mMCP 1. The integrin alphaE subunit is coexpressed with mMCP-1. CONCLUSION: The kinetics of mMCP-1+/alphaE+ mBMMC development, regulated by TGF-beta1, are consistent with that seen in vivo in the parasitized intestine. The normally down regulatory functions of TGF-beta1 in haematopoiesis are superseded in this culture system by its ability to promote the early expression of alphaE and mMCP 1. PMID- 11929500 TI - Antibodies directed against nerve growth factor inhibit the acute bronchoconstriction due to allergen challenge in guinea-pigs. AB - BACKGROUND: We have previously demonstrated that the administration of nerve growth factor (NGF) to guinea-pigs results in airway hyper-responsiveness within 1 h. OBJECTIVE: In the present study we document the involvement of NGF in the acute allergic airway response. METHODS: Guinea-pigs that are sensitized to ovalbumin show an acute bronchoconstriction directly after challenge with ovalbumin. RESULTS: Intratracheal application of 10 microg of antibodies directed against NGF (anti-NGF) 1 h before the challenge reduces the acute severe bronchoconstriction to approximately 40% and the sustained bronchoconstriction to approximately 20% of the reaction in controls. This shows a high potency of anti NGF in diminishing the direct bronchoconstriction. Inhibition of the tyrosine kinases of the tyrosine kinase receptor A, the high-affinity receptor for NGF, has no effect on the bronchoconstriction. Therefore, we postulate that the p75, the low-affinity receptor for neurotrophins, is responsible for the acute bronchoconstriction. Our findings suggest a role for NGF in the induction of the acute asthmatic reaction. CONCLUSION: These findings offer a new potential therapeutic strategy for the treatment of allergic asthma. PMID- 11929501 TI - Physician office visits of adults for anxiety disorders in the United States, 1985-1998. AB - OBJECTIVE: To determine the number of physician office visits by adults in which an anxiety disorder diagnosis was recorded and rates of treatment during these visits. DESIGN: We used data from the 1985, 1993, 1994, 1997, and 1998 National Ambulatory Medical Care Surveys, which is a nationally representative series of surveys of office-based practice employing clustered sampling. SETTING: Office based physician practices in the United States. PARTICIPANTS: A systematically sampled group of office-based physicians. RESULTS: The number of office visits with a recorded anxiety disorder diagnosis increased from 9.5 million in 1985 to 11.2 million per year in 1993-1994 and 12.3 million per year in 1997-1998, representing 1.9%, 1.6%, and 1.5% of all office visits in 1985, 1993-1994, and 1997-1998, respectively. The majority of recorded anxiety disorder diagnoses were not for specific disorders, with 70% of anxiety disorder visits to primary care physicians coded as "anxiety state, unspecified." Visits to primary care physicians accounted for 48% of all anxiety disorder visits in 1985 and 1997 1998. Treatment for anxiety was offered in over 95% of visits to psychiatrists but in only 60% of visits to primary care physicians. Primary care physicians were less likely to offer treatment for anxiety when specific anxiety disorders were diagnosed than when "anxiety state, unspecified" was diagnosed (54% vs 62% in 1997-1998). Prescriptions for medications to treat anxiety disorders increased between 1985 and 1997-1998 while use of psychotherapy decreased over the same time period in visits to both primary care physicians and psychiatrists. CONCLUSIONS: Although there is a large number of office visits with a recorded anxiety disorder diagnosis, under-recognition and under-treatment appear to be a continuing problem, especially in the primary care sector. Medication is being substituted for psychotherapy in visits to both psychiatrists and primary care physicians over time. PMID- 11929502 TI - Use of opioid medications for chronic noncancer pain syndromes in primary care. AB - OBJECTIVES: To define the spectrum of chronic noncancer pain treated with opioid medications in 2 primary care settings, and the prevalence of psychiatric comorbidity in this patient population. We also sought to determine the proportion of patients who manifested prescription opioid abuse behaviors and the factors associated with these behaviors. DESIGN: A retrospective cohort study. SETTING: A VA primary care clinic and an urban hospital-based primary care center (PCC) located in the northeastern United States. PATIENTS: A random sample of VA patients ( n=50) and all PCC patients ( n=48) with chronic noncancer pain who received 6 or more months of opioid prescriptions during a 1-year period (April 1, 1997 through March 31, 1998) and were not on methadone maintenance. MEASUREMENTS: Information regarding patients' type of chronic pain disorder, demographic, medical, and psychiatric status, and the presence of prescription opioid abuse behaviors was obtained by medical record review. MAIN RESULTS: Low back pain was the most common disorder accounting for 44% and 25% of all chronic pain diagnoses in the VA and PCC samples, respectively, followed by injury related (10% and 13%), diabetic neuropathy (8% and 10%), degenerative joint disease (16% and 13%), spinal stenosis (10% and 4%), headache (4% and 13%) and other chronic pain disorders (8% and 22%). The median duration of pain was 10 years (range 3 to 50 years) in the VA and 13 years in the PCC sample (range 1 to 49 years). Among VA and PCC patients, the lifetime prevalence rates of psychiatric comorbidities were: depressive disorder (44% and 54%), anxiety disorder (20% and 21%), alcohol abuse/dependence (46% and 31%), and narcotic abuse/dependence (18% and 38%). Prescription opioid abusive behaviors were recorded for 24% of VA and 31% of PCC patients. A lifetime history of a substance use disorder (adjusted odds ratio [OR], 3.8; 95% confidence interval [CI], 1.4 to 10.8) and age (adjusted OR, 0.94; 95% CI, 0.89 to 0.99) were independent predictors of prescription opioid abuse behavior. CONCLUSIONS: A broad spectrum of chronic noncancer pain disorders are treated with opioid medications in primary care settings. The lifetime prevalence of psychiatric comorbidity was substantial in our study population. A significant minority of patients manifested prescription opioid abusive behaviors, and a lifetime history of a substance use disorder and decreasing age were associated with prescription opioid abuse behavior. Prospective studies are needed to determine the potential benefits as well as risks associated with opioid use for chronic noncancer pain in primary care. PMID- 11929503 TI - Patients' use of the Internet for medical information. AB - OBJECTIVES: To determine the percentage of patients enrolled in a primary care practice who use the Internet for health information, to describe the types of information sought, to evaluate patients' perceptions of the quality of this information, and to determine if patients who use the Internet for health information discuss this with their doctors. DESIGN: Self-administered mailed survey. SETTING: Patients from a primary care internal medicine private practice. PARTICIPANTS: Randomly selected patients ( N=1,000) were mailed a confidential survey between December 1999 and March 2000. The response rate was 56.2%. MEASUREMENTS AND MAIN RESULTS: Of the 512 patients who returned the survey, 53.5% (274) stated that they used the Internet for medical information. Those using the Internet for medical information were more educated ( P <.001) and had higher incomes ( P <.001). Respondents used the Internet for information on a broad range of medical topics. Sixty percent felt that the information on the Internet was the "same as" or "better than" information from their doctors. Of those using the Internet for health information, 59% did not discuss this information with their doctor. Neither gender, education level, nor age less than 60 years was associated with patients sharing their Web searches with their physicians. However, patients who discussed this information with their doctors rated the quality of information higher than those who did not share this information with their providers. CONCLUSIONS: Primary care providers should recognize that patients are using the World Wide Web as a source of medical and health information and should be prepared to offer suggestions for Web-based health resources and to assist patients in evaluating the quality of medical information available on the Internet. PMID- 11929504 TI - Effect of discharge summary availability during post-discharge visits on hospital readmission. AB - OBJECTIVE: To determine if the delivery of hospital discharge summaries to follow up physicians decreases the risk of hospital readmission. SUBJECTS: Eight hundred eighty-eight patients discharged from a single hospital following treatment for an acute medical illness. SETTING: Teaching hospital in a universal health-care system. DESIGN: We determined the date that each patient's discharge summary was printed and the physicians to whom it was sent. Summary receipt was confirmed by survey and phoning each physician's office. Each patient's hospital chart was reviewed to determine their acute and chronic medical conditions as well as their course in hospital. Using population-based administrative databases, all post hospitalization visits were identified. For each of these visits, we determined whether the summary was available. MAIN OUTCOME MEASURES: Time to nonelective hospital readmission during 3 months following discharge. RESULTS: The discharge summary was available for only 568 of 4,639 outpatient visits (12.2%). Overall, 240 (27.0%) of patients were urgently readmitted to hospital. After adjusting for significant patient and hospitalization factors, we found a trend toward a decreased risk of readmission for patients who were seen in follow-up by a physician who had received a summary (relative risk 0.74, 95% confidence interval 0.50 to 1.11). CONCLUSIONS: The risk of rehospitalization may decrease when patients are assessed following discharge by physicians who have received the discharge summary. Further research is required to determine if better continuity of patient information improves patient outcomes. PMID- 11929506 TI - Reporting of numerical and statistical differences in abstracts: improving but not optimal. AB - OBJECTIVE: The reporting of relative risk reductions (RRRs) or absolute risk reductions (ARRs) to quantify binary outcomes in trials engenders differing perceptions of therapeutic efficacy, and the merits of P values versus confidence intervals (CIs) are also controversial. We describe the manner in which numerical and statistical difference in treatment outcomes is presented in published abstracts. DESIGN: A descriptive study of abstracts published in 1986 and 1996 in 8 general medical and specialty journals. INCLUSION CRITERIA: controlled, intervention trials with a binary primary or secondary outcome. Seven items were recorded: raw data (outcomes for each treatment arm), measure of relative difference (e.g., RRR), ARR, number needed to treat, P value, CI, and verbal statement of statistical significance. The prevalence of these items was compared between journals and across time. RESULTS: Of 5,293 abstracts, 300 met the inclusion criteria. In 1986, 60% of abstracts did not provide both the raw data and a corresponding P value or CI, while 28% failed to do so in 1Dr. Hux is a Career Scientist of the Ontario Ministry of Health and receives salary support from the Institute for Clinical Evaluative Sciences in Ontario.996 ( P <.001; RRR of 53%; ARR of 32%; CI for ARR 21% to 43%). The variability between journals was highly significant ( P <.001). In 1986, 100% of abstracts lacked a measure of absolute difference while 88% of 1996 abstracts did so ( P <.001). In 1986, 98% of abstracts lacked a CI while 65% of 1996 abstracts did so ( P <.001). CONCLUSIONS: The provision of quantitative outcome and statistical quantitative information has significantly increased between 1986 and 1996. However, further progress can be made to make abstracts more informative. PMID- 11929507 TI - Ambulatory morning report: can it prepare residents for the American Board of Internal Medicine Examination? AB - We assessed the ability of a novel ambulatory morning report format to expose internal medicine residents to the breadth of topics covered by the American Board of Internal Medicine (ABIM) exam. Cases were selected by the Ambulatory Assistant Chief Residents and recorded in a logbook to limit duplication. We conducted a retrospective review of 406 cases discussed from July 1998 to July 2000 and cataloged each according to the primary content area. The percentage of cases in each area accurately reflected that covered by the ABIM exam, with little redundancy or over-selection of esoteric diseases. Our data suggest that a general medicine clinic is capable of exposing house staff to the wide breadth of internal medicine topics previously thought to be unique to subspecialty clinics. PMID- 11929505 TI - Optimal strategy for the first episode of primary spontaneous pneumothorax in young men. A decision analysis. AB - OBJECTIVE: Primary spontaneous pneumothorax (PSP) is not uncommon in young men and is associated with frequent recurrence. The frequent recurrence after conservative treatment and resultant anxiety for recurrence are sources of disability. We explored which procedure is more appropriate as the initial therapy in terms of quality-adjusted life expectancy (QALE). DESIGN: Decision analysis using a Markov model. DATA SOURCES: Structured literature review for clinical probability. Utility derived from patients and medical staff using time trade-off method. SETTING: Hypothetical cohort. PATIENTS: Twenty-year-old men with a first episode of PSP for which simple aspiration was ineffective. INTERVENTIONS: One of the following treatment options: 1) thoracoscopic surgery, 2) pleural drainage followed by thoracoscopic surgery for recurrence, 3) pleural drainage followed by thoracoscopic surgery for the second recurrence, 4) pleurodesis followed by thoracoscopic surgery for recurrence, 5) pleurodesis followed by thoracoscopic surgery for the second recurrence, 6) pleural drainage followed by pleurodesis for the first recurrence and thoracoscopic surgery for the second recurrence. MEASUREMENTS AND MAIN RESULTS: During the 1-year period after one of the initial treatments, the QALE was 9.49 months for thoracoscopic surgery, 9.47 for pleurodesis, and 7.80-7.99 for pleural drainage. The QALE for thoracoscopic surgery was the longest among the 6 strategies during the period from 5 to 24 months. None of the variables in sensitivity analyses altered the main results except for thoracoscopic surgical death rate. When it exceeds 0.3%, pleurodesis becomes the preferred strategy. CONCLUSION: On the basis of the current best available data and patients' preference, thoracoscopic surgery can be considered the treatment of choice for the first episode of PSP. PMID- 11929509 TI - Social context of work injury among undocumented day laborers in San Francisco. AB - OBJECTIVE: To identify ways in which undocumented day laborers' social context affects their risk for occupational injury, and to characterize the ways in which these workers' social context influences their experience of disability. DESIGN: Qualitative study employing ethnographic techniques of participant-observation, supplemented by semistructured in-depth interviews. SETTINGS: Street corners in San Francisco's Mission District, a homeless shelter, and a nonprofit day labor hiring hall. PARTICIPANTS: Thirty-eight Mexican and Central American male day laborers, 11 of whom had been injured. PRIMARY THEMES: Anxiety over the potential for work injury is omnipresent for day laborers. They work in dangerous settings, and a variety of factors such as lack of training, inadequate safety equipment, and economic pressures further increase their risk for work injury. The day laborers are isolated from family and community support, living in a local context of homelessness, competition, and violence. Injuries tend to have severe emotional, social, and economic ramifications. Day laborers frequently perceive injury as a personal failure that threatens their masculinity and their status as patriarch of the family. Their shame and disappointment at failing to fulfill culturally defined masculine responsibilities leads to intense personal stress and can break family bonds. Despite the high incidence of work injuries and prevalence of work-related health conditions, day laborers are frequently reluctant to use health services due to anxiety regarding immigration status, communication barriers, and economic pressure. IMPLICATIONS: On the basis of these ethnographic data, we recommend strategies to improve ambulatory care services to day laborers in 3 areas: structural changes in ambulatory care delivery, clinical interactions with individual day laborers, and policymaking around immigration and health care issues. PMID- 11929508 TI - The limited incorporation of economic analyses in clinical practice guidelines. AB - BACKGROUND: Because there is increasing concern that economic data are not used in the clinical guideline development process, our objective was to evaluate the extent to which economic analyses are incorporated in guideline development. METHODS: We searched medline and HealthSTAR databases to identify English language clinical practice guidelines (1996-1999) and economic analyses (1990 1998). Additional guidelines were obtained from The National Guidelines Clearinghouse Internet site available at http://www.guideline.gov. Eligible guidelines met the Institute of Medicine definition and addressed a topic included in an economic analysis. Eligible economic analyses assessed interventions addressed in a guideline and predated the guideline by 1 or more years. Economic analyses were defined as incorporated in guideline development if 1) the economic analysis or the results were mentioned in the text or 2) listed as a reference. The quality of economic analyses was assessed using a structured scoring system. RESULTS: Using guidelines as the unit of analysis, 9 of 35 (26%) incorporated at least 1 economic analysis of above-average quality in the text and 11 of 35 (31%) incorporated at least 1 in the references. Using economic analyses as the unit of analysis, 63 economic analyses of above-average quality had opportunities for incorporation in 198 instances across the 35 guidelines. Economic analyses were incorporated in the text in 13 of 198 instances (7%) and in the references in 18 of 198 instances (9%). CONCLUSIONS: Rigorous economic analyses may be infrequently incorporated in the development of clinical practice guidelines. A systematic approach to guideline development should be used to ensure the consideration of economic analyses so that recommendations from guidelines may impact both the quality of care and the efficient allocation of resources. PMID- 11929510 TI - Antibiotic treatment of acute bronchitis in smokers: a systematic review. AB - OBJECTIVE: Community physicians in the United States prescribe antibiotics to 80% to 90% of smokers with acute bronchitis. We performed a systematic review of the literature to determine the efficacy of antibiotics for smokers with acute bronchitis. DESIGN: A medline search was done using the keywords bronchitis, cough, and antibiotics to identify English language articles published from January 1966 to September 2001. Randomized, placebo-controlled trials of antibiotics in previously healthy smokers and nonsmokers with acute bronchitis were included. MEASUREMENTS AND MAIN RESULTS: For each study, we abstracted information on design, size, inclusion criteria, patient characteristics, and outcomes. Of 2,029 articles in the original search, 109 relevant articles were retrieved and reviewed. There have been no studies specifically addressing antibiotic use in smokers with acute bronchitis. Nine randomized, placebo controlled trials of antibiotics have included 774 patients and over 276 smokers. Lack of subgroup reporting for smokers precluded meta-analysis. In 7 trials, smoking status did not predict or alter patients' response to antibiotics. In one trial, trimethoprim/sulfamethoxazole resulted in less-frequent cough overall, but not among smokers. In another trial, erythromycin reduced symptom scores only among nonsmokers while antibiotic-treated smokers had a trend toward higher symptom scores. CONCLUSION: Although no trials have specifically addressed antibiotic use in smokers with acute bronchitis, existing data suggest that any benefit of antibiotics is the same or less for smokers than for nonsmokers. PMID- 11929511 TI - Economic analyses and clinical practice guidelines: why not a match made in heaven? PMID- 11929512 TI - Chronic pain and narcotics: a dilemma for primary care. PMID- 11929513 TI - Proteolysis and its regulation at the surface of Streptococcus pyogenes. AB - Pathogenic bacteria often produce proteinases that are believed to be involved in virulence. Moreover, several host defence systems depend on proteolysis, demonstrating that proteolysis and its regulation play an important role during bacterial infections. Here, we discuss how proteolytical events are regulated at the surface of Streptococcus pyogenes during infection with this important human pathogen. Streptococcus pyogenes produces proteinases, and host proteinases are produced and released as a result of the infection. Streptococcus pyogenes also recruits host proteinase inhibitors to its surface, suggesting that proteolysis is tightly regulated at the bacterial surface. We propose that the initial phase of a S. pyogenes infection is characterized by inhibition of proteolysis and complement activity at the bacterial surface. This is achieved mainly through binding of host proteinase inhibitors and complement regulatory proteins to bacterial surface proteins. In a later phase of the infection, massive proteolytic activity will release bacterial surface proteins and degrade human tissues, thus facilitating bacterial spread. These proteolytic events are regulated both temporally and spatially, and should influence virulence and the outcome of S. pyogenes infections. PMID- 11929514 TI - The yeast transcriptome in aerobic and hypoxic conditions: effects of hap1, rox1, rox3 and srb10 deletions. AB - The transcriptome of Saccharomyces cerevisiae was screened using the high-density membrane hybridization method, under aerobic and hypoxic conditions, in wild-type and mutant backgrounds obtained by the disruption of the genes encoding the regulatory proteins Hap1, Rox1 and the Srb10 and Rox3 subunits of RNA polymerase II holoenzyme. None of the mutations studied was able to fully overcome the wild type hypoxic response. Deletion of the hap1 gene changed the expression profiles of individual open reading frames (ORFs) under both aerobic and hypoxic conditions. Major changes associated with rox3 deletion were related to the hypoxic activation. Rox3 also caused a repressor effect (oxygen-independent) on a subset of genes related to subtelomeric proteins. With regard to the effect brought about by the deletion of rox1 and srb10, correspondence cluster analysis revealed that the transcriptome profile in aerobic conditions is very similar in the wild-type and both deletion strains. In contrast, however, differences were found during hypoxia between the subgroup formed by wild-type and the Deltarox1 deletant compared with the Deltasrb10 deletant. An analysis of selected ORFs responding to hypoxia, in association with a dependence on the regulatory factors studied, made it possible to identify the clusters that are related to different regulatory circuits. PMID- 11929515 TI - InlA- but not InlB-mediated internalization of Listeria monocytogenes by non phagocytic mammalian cells needs the support of other internalins. AB - To determine the contribution of the previously identified internalins, InlA, InlB, InlC, InlE, InlG, and InlH, to internalization of Listeria monocytogenes by non-professional phagocytic mammalian cells, we constructed mutants with various combinations of deletions in the respective inl genes. Internalization of these mutants into the epithelial-like Caco-2 and the microvascular endothelial HBMEC cell lines were studied. Deletion of the inlGHE gene cluster, or of the single genes, led to a two to fourfold increased internalization by HBMEC and other non phagocytic mammalian cells. Invasion into HBMEC was totally blocked in the absence of InlB, and InlB-dependent internalization did not require the presence of any of the other internalins. Internalization by Caco-2 cells was reduced to a level of about 1% in the absence of InlA and InlB, and was most efficient in the presence of InlA, InlB and InlC and in the absence of InlG, InlH and InlE. InlB and InlA, in each case in the absence of the other internalins, led (compared with the wild-type strain) to reduced internalization of about 20% and less than 10% respectively. InlA-dependent internalization (in the absence of InlB) required the additional function of InlC and InlGHE. The deletion of inlGHE enhanced the expression of InlA and InlB. The increased amount of InlA led to an increase in early association of L. monocytogenes with Caco-2 cells without enhancing its uptake in the absence of the other internalins, whereas the larger amount of InlB did not enhance early association of L. monocytogenes with HBMEC but led to an increase in internalization of L. monocytogenes. The results suggest that InlB is able to induce phagocytosis in HBMEC and (at a lower efficiency) in Caco-2 cells by itself, but InlA needs the supportive functions of the other internalins to trigger phagocytosis. None of these internalins seems to be required for cell-to-cell spread by L. monocytogenes, as shown by microinjection of Caco-2 cells with appropriate inl mutants. PMID- 11929516 TI - Functional analysis of a vacuolar ABC transporter in wild-type Candida albicans reveals its involvement in virulence. AB - ATP-driven transport proteins belonging to the ATP-binding cassette (ABC) superfamily perform important functions in cell metabolism and detoxification. Compounds can be actively transported across membranes, including the plasma membrane or organellar membranes. The vacuole is an important organelle in fungal cells required for compartmentalization of metabolites as well as toxic substances. Sequestration into the vacuole is often energy-dependent. We present the first isolation and molecular analysis of a vacuolar ABC transporter gene in the opportunistic fungal pathogen Candida albicans. The protein encoded by the MLT1 gene is highly similar to Multiple Drug Resistance-associated Protein (MRP) like transporters of yeast and higher organisms that form the Cystic Fibrosis Transmembrane Conductance Regulator (CFTR)/MRP subfamily of ABC transporters, a class of proteins so far not characterized in C. albicans. MLT1 expression is extensively growth phase-regulated, and gene transcripts are inducible by metabolic poisons. Gene replacement mutants generated in wild-type C. albicans with the dominant selection marker MPAR showed a profound reduction in virulence in a mouse peritonitis model that was reversed by complementation with an intact MLT1 gene. Hence, this report provides primary evidence for the involvement of vacuolar ABC transporters in fungal virulence. PMID- 11929517 TI - A putative three-dimensional targeting motif of polygalacturonase (PehA), a protein secreted through the type II (GSP) pathway in Erwinia carotovora. AB - Intramolecular information specifying protein secretion through the type II (GSP) pathway of Gram-negative bacteria was investigated. Two regions of the polygalacturonase (PehA) of Erwinia carotovora containing residues proposed to be included in a targeting motif were located, one close to the C-terminus between residues 342 and 369 and another between residues 84 and 135 in the large central loops. The regions were required together to promote secretion. Further residues in the middle of the protein were required for proper positioning of the regions, suggesting that they were both involved in interaction with the GSP. To our knowledge, this is the first time that a possible three-dimensional targeting motif has been defined. At least one of the motifs comprises a cluster on the surface of the protein. The two motifs are structurally dissimilar, suggesting that there are two distinct recognition regions in the GSP apparatus. Finally, we propose that the targeting motifs are of a complex conformational nature with some variability accommodated, as illustrated by the observation that many mutations exhibited no clear phenotype individually but, in combination, severely compromised secretion. PMID- 11929518 TI - Mutations in FlbD that relieve the dependency on flagellum assembly alter the temporal and spatial pattern of developmental transcription in Caulobacter crescentus. AB - The transcription factor FlbD regulates the temporal and spatial transcription of flagellar genes in the bacterium Caulobacter crescentus. Activation of FlbD requires cell cycle progression and the assembly of an early (class II) flagellum structure. In this report, we identify 20 independent gain-of-function mutations in flbD that relieve regulation by flagellar assembly. One of these, flbD-1204, contained a mutation in the receiver domain (V17M) and another, flbD-1231, in the DNA binding domain (V451G). Both of these mutations resulted in an aberrant pattern of cell cycle transcription. The presence of the FlbD-1204 allele also resulted in a loss of swarmer-pole-specific transcription. These results indicate that temporal and spatial transcription is influenced by the assembly of the nascent flagellar structure. The trans-acting positive and negative regulatory factor, FliX, couples flagellar assembly to the activation of FlbD and, as we show here, also influences temporal transcription. Furthermore, we show that FliX can suppress the activity of FlbD mutants that cannot be phosphorylated, and that FliX is required for FlbD stability, and vice versa. These results indicate that FliX may interact directly with FlbD to regulate its activity. PMID- 11929519 TI - Replication restart in UV-irradiated Escherichia coli involving pols II, III, V, PriA, RecA and RecFOR proteins. AB - In Escherichia coli, UV-irradiated cells resume DNA synthesis after a transient inhibition by a process called replication restart. To elucidate the role of several key proteins involved in this process, we have analysed the time dependence of replication restart in strains carrying a combination of mutations in lexA, recA, polB (pol II), umuDC (pol V), priA, dnaC, recF, recO or recR. We find that both pol II and the origin-independent primosome-assembling function of PriA are essential for the immediate recovery of DNA synthesis after UV irradiation. In their absence, translesion replication or 'replication readthrough' occurs approximately 50 min after UV and is pol V-dependent. In a wild-type, lexA+ background, mutations in recF, recO or recR block both pathways. Similar results were obtained with a lexA(Def) recF strain. However, lexA(Def) recO or lexA(Def) recR strains, although unable to facilitate PriA-pol II dependent restart, were able to perform pol V-dependent readthrough. The defects in restart attributed to mutations in recF, recO or recR were suppressed in a recA730 lexA(Def) strain expressing constitutively activated RecA (RecA*). Our data suggest that in a wild-type background, RecF, O and R are important for the induction of the SOS response and the formation of RecA*-dependent recombination intermediates necessary for PriA/Pol II-dependent replication restart. In con trast, only RecF is required for the activation of RecA that leads to the formation of pol V (UmuD'2C) and facilitates replication readthrough. PMID- 11929520 TI - Involvement of surface polysaccharides in the organic acid resistance of Shiga Toxin-producing Escherichia coli O157:H7. AB - In general, wild Escherichia coli strains can grow effectively under moderately acidic organic acid-rich conditions. We found that the Shiga Toxin-producing E. coli (STEC) O157:H7 NGY9 grows more quickly than a K-12 strain in Luria-Bertani (LB)-2-morpholinoethanesulphonic acid (MES) broth supplemented with acetic acid (pH 5.4). Hypothesizing that the resistance of STEC O157:H7 to acetic acid is as a result of a mechanism(s) other than those known, we screened for STEC mutants sensitive to acetic acid. NGY9 was subjected to mini-Tn5 mutagenesis and, from 50,000 colonies, five mutants that showed a clear acetic acid-sensitive phenotype were isolated. The insertion of mini-Tn5 in three mutants occurred at the fcl, wecA (rfe) and wecB (rffE) genes and caused loss of surface O-polysaccharide, loss of both O-polysaccharide and enterobacterial common antigen (ECA) and loss of ECA respectively. The other two mutants showed inactivation of the waaG (rfaG) gene but at different positions that caused a deep rough mutant with loss of the outer core oligosaccharide of lipopolysaccharide (LPS) as well as phenotypic loss of O-polysaccharide and ECA. With the introduction of plasmids carrying the fcl, wecA, wecB and waaG genes, respectively, all mutants were complemented in their production of O-polysaccharide and ECA, and normal growth was restored in organic acid-rich culture conditions. We also found that the growth of Salmonella LPS mutants Ra, Rb1, Rc, Rd1, Rd2 and Re was suppressed in the presence of acetic acid compared with that of the parents. These results suggest that the full expression of LPS (including O-polysaccharide) and ECA is indispensable to the resistance against acetic acid and other short chain fatty acids in STEC O157:H7 and Salmonella. To the best of our knowledge, this is a newly identified physiological role for O-polysaccharide and ECA as well as an acid resistance mechanism. PMID- 11929521 TI - Translational readthrough of the PDE2 stop codon modulates cAMP levels in Saccharomyces cerevisiae. AB - The efficiency of translation termination in yeast can vary several 100-fold, depending on the context around the stop codon. We performed a computer analysis designed to identify yeast open reading frames (ORFs) containing a readthrough motif surrounding the termination codon. Eight ORFs were found to display inefficient stop codon recognition, one of which, PDE2, encodes the high-affinity cAMP phosphodiesterase. We demonstrate that Pde2p stability is very impaired by the readthrough-dependent extension of the protein. A 20-fold increase in readthrough of PDE2 was observed in a [PSI+] as compared with a [psi-] strain. Consistent with this observation, an important increase in cAMP concentration was observed in suppressor backgrounds. These results provide a molecular explanation for at least some of the secondary phenotypes associated with suppressor backgrounds. PMID- 11929522 TI - Functional genomics reveals the sole sulphate transporter of the Mycobacterium tuberculosis complex and its relevance to the acquisition of sulphur in vivo. AB - Sulphur is essential for some of the most vital biological activities such as translation initiation and redox maintenance, and genes involved in sulphur metabolism have been implicated in virulence. Mycobacterium tuberculosis has three predicted genes for the prototrophic acquisition of sulphur as sulphate: cysA, part of an ABC transporter, and cysA2 and A3, SseC sulphotransferases. Screening for amino acid auxotrophs of Mycobacterium bovis BCG, obtained by transposon mutagenesis, was used to select methionine auxotrophs requiring a sulphur-containing amino acid for growth. We have characterized one of these auxotrophs as being disrupted in cysA. Both the cysA mutant and a previously identified mutant in an upstream gene, subI, were functionally characterized as being completely unable to take up sulphate. Complementation of the cysA mutant with the wild-type gene from M. tuberculosis restored prototrophy and the ability to take up sulphate with the functional characteristics of an ABC transporter. Hence, it appears that this is the sole locus encoding inorganic sulphur transport in the M. tuberculosis complex. PMID- 11929523 TI - Characterization of a bi-directional promoter for divergent transcription of a PHD-zinc finger protein gene and a ran gene in the protozoan pathogen Giardia lamblia. AB - We showed previously that transcription of the ran gene in Giardia lamblia is regulated by an AT-rich initiator. In the present study, the ran initiator was found to regulate transcription of a neighbouring PHD zinc-finger protein gene. Deletion and scanning mutagenesis of the phd promoter in a firefly luciferase reporter system showed that the promoter activity is determined by multiple single-stranded T-tract DNA elements distributed into a distal domain spanning the ran initiator (-134/-103) and a proximal domain (-88/-48) spanning phd messenger RNA (mRNA) start sites (-74, -55 and -53 relative to the first ATG). The promoter activity is repressed by the single T-tract element on a non template strand of the ran initiator, and is activated by closely spaced T-tract elements on the opposite strand. The T-tract elements in the phd and ran initiators compete for similar ssDNA binding proteins. Mutation of -47/-42 resulted in dramatic reduction of luciferase activity without changing luciferase mRNA levels, indicating the potential involvement of a regulatory mechanism in PHD protein translation. These findings suggest that G. lamblia uses multiple copies of a T-tract element as both core and distal elements in regulating transcription initiation, and that expression of the phd gene is regulated at multiple levels. PMID- 11929524 TI - Control of the AcrAB multidrug efflux pump by quorum-sensing regulator SdiA. AB - SdiA is an Escherichia coli protein that regulates cell division in a cell density-dependent, or quorum-sensing, manner. We report that SdiA also controls multidrug resistance by positively regulating the multidrug resistance pump AcrAB. Overproduction of SdiA confers multidrug resistance and increased levels of AcrAB. Conversely, sdiA null mutants are hypersensitive to drugs and have decreased levels of AcrB protein. Our findings provide a link between quorum sensing and multidrug efflux. Combined with previously published reports, our data support a model in which a role of drug efflux pumps is to mediate cell-cell communication in response to cell density. Xenobiotics expelled by pumps may resemble the communication molecules that they normally efflux. PMID- 11929525 TI - Independent regulation of the divergent Escherichia coli nrfA and acsP1 promoters by a nucleoprotein assembly at a shared regulatory region. AB - Expression from the Escherichia coli nrfA promoter (pnrfA) is activated by both the FNR protein (an anaerobically triggered transcription activator) and the NarL or NarP proteins (transcription activators triggered by nitrite and nitrate). Under anaerobic conditions, FNR binds to a site centred at position -41.5 at pnrfA and activates transcription. Further activation, induced by the presence of nitrite, results from the binding of NarL and NarP to a site centred at position 74.5. A second promoter (pacsP1), which directs transcription into the adjacent gene encoding acetyl coenzyme A synthetase (acs), is overlapping and divergent to pnrfA. Despite extensive overlap of regulatory elements, pnrfA and pacsP1 are regulated independently. We demonstrate that at least two nucleoid-associated factors bind to the nrfA-acs intergenic region. The Fis protein binds to a site centred at position -15 (in relation to pnrfA transcription), whereas the IHF protein binds to a site centred at position -54. Both Fis and IHF repress in vivo expression from pacsP1, but have smaller repressive effects on expression from pnrfA. Gel retardation assays were used to investigate the pairwise binding of FNR, NarL, Fis and IHF proteins to the nrfA-acs intergenic region. The binding of NarL and IHF is mutually exclusive, whereas all other combinations can bind simultaneously. Experiments in which deletions and point mutations were introduced into the upstream region of pnrfA demonstrated that an additional factor must bind upstream to inhibit FNR-dependent transcription. We conclude that the nrfA-acs intergenic region is folded into an ordered nucleoprotein structure that permits the two divergent promoters to be regulated independently in response to different physiological signals. PMID- 11929526 TI - An ABC-type, high-affinity urea permease identified in cyanobacteria. AB - Urea is an important nitrogen source for many microorganisms, but urea active transporters have not been characterized at a molecular level in any bacterium. Cells of Synechocystis sp. PCC 6803 and Anabaena sp. PCC 7120 exhibited the capacity to take up [14C]-urea from low-concentration (<1 microM) urea solutions. The Ks of Anabaena cells for urea was about 0.11 microM, and the observed uptake activity involved the transport and metabolism of urea. In contrast to urease, which was constitutively ex-pressed, expression of the high-affinity urea uptake activity was subjected to nitrogen control. In an Anabaena ureG (urease-) mutant, a concentrative, active transport of urea could be demonstrated. We found that a mutant of open reading frame (ORF) sll0374 from the Synechocystis genomic sequence lacked urea transport activity. This ORF encoded a conserved component of an ABC-type transporter, but it is not clustered together with any other possible transporter-encoding gene. An Anabaena homologue of sll0374, urtE, was isolated and found to be part of a cluster of genes, urtABCDE, putatively encoding all the elements of an ABC-type permease. Although the longest transcript that we could detect only covered urtABC, the impairment of urea transport by inactivation of urtA, urtB or urtE suggested that the whole gene cluster is expressed producing the urea permease. Expression was induced under nitrogen-limiting conditions, and a complex promoter regulated by the cyanobacterial global nitrogen control transcription factor NtcA was found upstream from urtA. Our work adds urea to the known substrates of the versatile class of ABC-type transporters and suggests the involvement of a transporter of this superfamily in urea scavenging by some bacteria in natural environments. PMID- 11929527 TI - Evaluation of a nutrient starvation model of Mycobacterium tuberculosis persistence by gene and protein expression profiling. AB - The search for new TB drugs that rapidly and effectively sterilize the tissues and are thus able to shorten the duration of chemotherapy from the current 6 months has been hampered by a lack of understanding of the metabolism of the bacterium when in a 'persistent' or latent form. Little is known about the condition in which the bacilli survive, although laboratory models have shown that Mycobacterium tuberculosis can exist in a non-growing, drug-resistant state that may mimic persistence in vivo. Using nutrient starvation, we have established a model in which M. tuberculosis arrests growth, decreases its respiration rate and is resistant to isoniazid, rifampicin and metronidazole. We have used microarray and proteome analysis to investigate the response of M. tuberculosis to nutrient starvation. Proteome analysis of 6-week-starved cultures revealed the induction of several proteins. Microarray analysis enabled us to monitor gene expression during adaptation to nutrient starvation and confirmed the changes seen at the protein level. This has provided evidence for slowdown of the transcription apparatus, energy metabolism, lipid biosynthesis and cell division in addition to induction of the stringent response and several other genes that may play a role in maintaining long-term survival within the host. Thus, we have generated a model with which we can search for agents active against persistent M. tuberculosis and revealed a number of potential targets expressed under these conditions. PMID- 11929528 TI - H-NS-dependent activation of pectate lyases synthesis in the phytopathogenic bacterium Erwinia chrysanthemi is mediated by the PecT repressor. AB - Production of the main virulence determinant pectate lyases (Pels) of the phytopathogenic bacterium Erwinia chrysanthemi is modulated by a complex regulatory network involving the repressor proteins KdgR, PecS and PecT and the activator systems Pir, ExpI-ExpR and CRP. Of these regulators, CRP and PecT are particularly important since the absence of CRP or a slight overproduction of PecT leads to a drastic reduction in synthesis of Pel species. Recently, it has been shown that production of Pel species is strongly reduced in an E. chrysanthemi hns mutant, suggesting an activator function of the nucleoid associated protein H-NS in the expression of the pel genes. Here, we report that the reduced synthesis of Pel species in the hns mutant results from a negative control, exerted by H-NS, on the transcription of the regulatory gene pecT. This H-NS/PecT cascade regulation is one of the first elucidations of a positive effect of H-NS on target gene expression. Moreover, we found that H-NS also represses the expression of expI, expR and pel genes. H-NS control is the result of H-NS binding to extended regions within the pecT, expI, expR and pel genes. Investigation of the simultaneous binding of CRP, RNA polymerase (RNAP) and H-NS on the pelD gene revealed that these three proteins form a nucleoprotein com plex. Together, these data indicate that, by exerting a negative control at multiple levels, H-NS plays a crucial role in the E. chrysanthemi pel regulatory network. PMID- 11929529 TI - Co-ordinated expression of phycobiliprotein operons in the chromatically adapting cyanobacterium Calothrix PCC 7601: a role for RcaD and RcaG. AB - In the cyanobacterium Calothrix sp. PCC 7601 the cpc2 operon encoding phycocyanin 2 (PC2) is expressed if red radiations are available. RcaD was previously identified in extracts from red-light-grown cells as an alkaline phosphatase sensitive protein that binds upstream of the transcription start point (TSP) of the cpc2 operon. In this work, RcaD was purified, and the corresponding gene cloned with a PCR probe obtained using degenerated primers based on RcaD peptide sequences (accession no. AJ319541). Purified RcaD binds to the cpc2 promoter region and also to those of the constitutive cpc1 and apc1 operons that encode phycocyanin 1 and allophycocyanin. Escherichia coli-overexpressed RcaD can bind to the cpc2 promoter region. The rcaD gene is upstream of an open reading frame (ORF) termed rcaG. Co-transcription of both genes was demonstrated by reverse transcription (RT)-PCR experiments, and found to be independent of the light wavelengths. A single TSP was mapped. Sequence features of RcaD and RcaG led us to propose a functional relationship between these two proteins. A rcaD mutant generated by allelic exchange exhibited altered expression of the cpc2, cpeBA, apc1 and cpc1 operons upon green to red-light shifts. RcaD seems to be a co activator co-ordinating the transcription of the phycobiliprotein operons upon changes in light spectral quality. PMID- 11929530 TI - The nrfI gene is essential for the attachment of the active site haem group of Wolinella succinogenes cytochrome c nitrite reductase. AB - The cytochrome c nitrite reductase complex (NrfHA) is the terminal enzyme in the electron transport chain catalysing nitrite respiration of Wolinella succinogenes. The catalytic subunit NrfA is a pentahaem cytochrome c containing an active site haem group which is covalently bound via the cysteine residues of a unique CWTCK motif. The lysine residue serves as the axial ligand of the haem iron. The other four haem groups of NrfA are bound by conventional haem-binding motifs (CXXCH). The nrfHAIJ locus was restored on the genome of the W. succinogenes DeltanrfAIJ deletion mutant by integration of a plasmid, thus enabling the expression of modified alleles of nrfA and nrfI. A mutant (K134H) was constructed which contained a nrfA gene encoding a CWTCH motif instead of CWTCK. NrfA of strain K134H was found to be synthesized with five bound haem groups, as judged by matrix-assisted laser-desorption/ionization (MALDI) mass spectrometry. Its nitrite reduction activity with reduced benzyl viologen was 40% of the wild-type activity. Ammonia was formed as the only product of nitrite reduction. The mutant did not grow by nitrite respiration and its electron transport activity from formate to nitrite was 5% of that of the wild-type strain. The predicted nrfI gene product is similar to proteins involved in system II cytochrome c biogenesis. A mutant of W. succinogenes (stopI) was constructed that contained a nrfHAIJ gene cluster with the nrfI codons 47 and 48 altered to stop codons. The NrfA protein of this mutant did not catalyse nitrite reduction and lacked the active site haem group, whereas the other four haem groups were present. Mutant (K134H/stopI) which contained the K134H modification in NrfA in addition to the inactivated nrfI gene had essentially the same properties as strain K134H. NrfA from strain K134H/stopI contained five haem groups. It is concluded that NrfI is involved in haem attachment to the CWTCK motif in NrfA but not to any of the CXXCH motifs. The nrfI gene is obviously dispensable for maturation of a modified NrfA protein containing a CWTCH motif instead of CWTCK. Therefore, NrfI might function as a specific haem lyase that recognizes the active site lysine residue of NrfA. A similar role has been proposed for NrfE, F and G of Escherichia coli, although these proteins share no overall sequence similarity to NrfI and belong to system I cytochrome c biogenesis, which differs fundamentally from system II. PMID- 11929531 TI - The alternative sigma factor sigmaE controls antioxidant defences required for Salmonella virulence and stationary-phase survival. AB - Bacteria must contend with conditions of nutrient limitation in all natural environments. Complex programmes of gene expression, controlled in part by the alternative sigma factors sigmaS (sigma38, RpoS) and sigmaH (sigma32, RpoH), allow a number of bacterial species to survive conditions of partial or complete starvation. We show here that the alternative sigma factor sigmaE (sigma24, RpoE) also facilitates the survival of Salmonella typhimurium under conditions of nutrient deprivation. Expression of the sigmaE regulon is strongly induced upon entry of Salmonella into stationary phase. A Salmonella mutant lacking sigmaE has reduced survival during stationary phase as well as increased susceptibility to oxidative stress. A Salmonella strain lacking both sigmaE and sigmaS is non viable after just 24 h in stationary phase, but survival of these mutants is completely preserved under anaerobic stationary-phase conditions, suggesting that oxidative injury is one of the major mechanisms of reduced microbial viability during periods of nutrient deprivation. Moreover, the attenuated virulence of sigmaE-deficient Salmonella for mice can be largely restored by genetic abrogation of the host phagocyte respiratory burst, suggesting that the sigmaE regulon plays an important antioxidant role during Salmonella infection of mammalian hosts. PMID- 11929532 TI - Molecular analysis of KTI12/TOT4, a Saccharomyces cerevisiae gene required for Kluyveromyces lactis zymocin action. AB - TOT, the putative Kluyveromyces lactis zymocin target complex from Saccharomyces cerevisiae, is encoded by TOT1-7, six loci of which are isoallelic to RNA polymerase II (RNAPII) Elongator genes (ELP1-6). Unlike TOT1-3 (ELP1-3) and TOT5 7 (ELP5, ELP6 and ELP4 respectively), which display zymocin resistance when deleted, TOT4 (KTI12) also renders cells refractory to zymocin when maintained in multicopy or overexpressed from the GAL10 promoter. Elevated TOT4 copy number results in an intermediate tot phenotype, which includes mild sensitivities towards caffeine, Calcofluor white and elevated growth temperature, suggesting that TOT4 influences TOT/Elongator function. Tot4p interacts with Elongator, as shown by co-immunoprecipitation, and cell fractionation studies demonstrate partial co-migration with RNAPII and Elongator. As Elongator subunit interaction is not affected by either deletion of TOT4 or multicopy TOT4, Tot4p may not be a structural Elongator subunit but, rather, may regulate TOT/Elongator in a fashion that requires transient physical contact with TOT/Elongator. Consistent with a regulatory role, the presence of a potential P-loop motif conserved between yeast and human TOT4 homologues suggests capability of ATP or GTP binding and P-loop deletion renders Tot4p biologically inactive. PMID- 11929533 TI - Genetic analysis of Salmonella enteritidis biofilm formation: critical role of cellulose. AB - We report here a new screening method based on the fluorescence of colonies on calcofluor agar plates to identify transposon insertion mutants of Salmonella enteritidis that are defective in biofilm development. The results not only confirmed the requirement of genes already described for the modulation of multicellular behaviour in Salmonella typhimurium and other species, but also revealed new aspects of the biofilm formation process, such as two new genetic elements, named as bcsABZC and bcsEFG operons, required for the synthesis of an exopolysaccharide, digestible with cellulase. Non-polar mutations of bcsC and bcsE genes and complementation experiments demonstrated that both operons are responsible for cellulose biosynthesis in both S. enteritidis and S. typhimurium. Using two different growth media, ATM and LB, we showed that the biofilm produced by S. enteritidis is made of different constituents, suggesting that biofilm composition and regulation depends on environmental conditions. Bacterial adherence and invasion assays of eukaryotic cells and in vivo virulence studies of cellulose-deficient mutants indicated that, at least under our experimental conditions, the production of cellulose is not involved in the virulence of S. enteritidis. However, cellulose-deficient mutants were more sensitive to chlorine treatments, suggesting that cellulose production and biofilm formation may be an important factor for the survival of S. enteritidis on surface environments. PMID- 11929534 TI - Quorum sensing Escherichia coli regulators B and C (QseBC): a novel two-component regulatory system involved in the regulation of flagella and motility by quorum sensing in E. coli. AB - Quorum sensing is a cell-to-cell signalling mechanism in which bacteria secrete hormone-like compounds called autoinducers. When these auto-inducers reach a certain threshold concentration, they interact with bacterial transcriptional regulators, thereby regulating gene expression. Enterohaemorrhagic Escherichia coli (EHEC) O157:H7 as well as E. coli K-12 produces the autoinducer-2 (AI-2), which is synthesized by the product of the luxS gene, and previous work from our laboratory has shown that genes encoding the EHEC type III secretion system were activated by quorum sensing. Recently, by hybridizing an E. coli K-12 gene array with cDNA synthesized from RNA extracted from EHEC strain 86-24 and its isogenic luxS mutant, we observed that other potential virulence-associated factors, such as genes encoding the expression and assembly of flagella, motility and chemotaxis, were also activated by quorum sensing. The array data also indicated that several genes encoding putative E. coli regulators were controlled by quorum sensing. In this report, we describe a two-component system regulated by quorum sensing that shares homology with Salmonella typhimurium PmrAB, which we have named quorum sensing E. coli regulator B and C (QseBC). The qseBC genes, previously identified only as open reading frames b3025 and b3026, are organized in an operon in the E. coli chromosome, with qseB encoding the response regulator and qseC the sensor kinase. We confirmed the regulation of qseBC by quorum sensing using qseB::lacZ transcriptional fusions and characterized the phenotypes of an isogenic qseC mutation in EHEC. This mutant expressed less flagellin and had reduced motility compared with the wild-type and complemented strains. Transcription of flhD, fliA, motA and fliC::lacZ fusions was decreased in the qseC mutant, suggesting that qseBC is a transcriptional regulator of flagella genes. A qseC mutant was also generated in E. coli K-12 strain MC1000 that showed the same phenotypes as the EHEC mutant, indicating that qseBC regulates flagella and motility by quorum sensing in both EHEC and K-12. QseBC activates transcription of flhDC, which is the master regulator for the flagella and motility genes and, in the absence of flhD, QseBC failed to activate the transcription of fliA. Motility of a luxS, but not of a qseC, mutant can be restored by providing AI-2 exogenously as preconditioned media, suggesting that the qseC mutant is unable to respond to AI-2. However, QseC has no effect on the expression of other quorum sensing-controlled genes such as those encoding for the type III secretion system. These data indicate that QseBC is one component of the quorum-sensing regulatory cascade in both EHEC and K-12 that is involved in the regulation of flagella and motility genes, but that additional regulators in this cascade remain to be characterized. PMID- 11929535 TI - A novel family of regulated helicases/nucleases from Gram-positive bacteria: insights into the initiation of DNA recombination. PMID- 11929536 TI - Protein phosphatase 2A on track for nutrient-induced signalling in yeast. AB - Early studies identified two bona fide protein phosphatase 2A (PP2A)-encoding genes in Saccharomyces cerevisiae, designated PPH21 and PPH22. In addition, three PP2A-related phosphatases, encoded by PPH3, SIT4 and PPG1, have been identified. All share as much as 86% sequence similarity at the amino acid level. This review will focus primarily on Pph21 and Pph22, but some aspects of Sit4 regulation will also be discussed. Whereas a role for PP2A in yeast morphology and cell cycle has been readily recognized, uncovering its function in yeast signal transduction is a more recent breakthrough. Via their interaction with phosphorylated Tap42, PP2A and Sit4 play a pivotal role in target of rapamycin (TOR) signalling. PPH22 overexpression mimics overactive cAMP-PKA (protein kinase A) signalling and PP2A and Sit4 might represent ceramide signalling targets. The methylation of its catalytic subunit stabilizes the heterotrimeric form of PP2A and might counteract TOR signalling. We will show how these new elements could lead us to understand the role and regulation of PP2A in nutrient-induced signalling in baker's yeast. PMID- 11929537 TI - Phage P4 origin-binding domain structure reveals a mechanism for regulation of DNA-binding activity by homo- and heterodimerization of winged helix proteins. AB - The origin-binding domain of the gpalpha protein of phage P4 (P4-OBD) mediates origin recognition and regulation of gpalpha activity by the protein Cnr. We have determined the crystal structure of P4-OBD at 2.95 A resolution. The structure of P4-OBD is that of a dimer with pseudo twofold symmetry. Each subunit has a winged helix topology with a unique structure among initiator proteins. The only structural homologue of the P4-OBD subunit is the DNA-binding domain of the eukaryotic transcriptional activator Rfx1. Based on this structural alignment, a model for origin recognition by the P4-OBD dimer is suggested. P4-OBD mutations that interfere with Cnr binding locate to the dimer interface, indicating that Cnr acts by disrupting the gpalpha dimer. P4-OBD dimerization is mediated by helices alpha1 and alpha3 in both subunits, a mode of winged helix protein dimerization that is reminiscent of that of the eukaryotic transcription factors E2F and DP. This, in turn, suggests that Cnr is also a winged helix protein, a possibility that is supported by previously unreported sequence homologies between Cnr and Rfx1 and homology modelling. Hence, in a mechanism that appears to be conserved from phage to man, the DNA-binding activity of winged helix proteins can be regulated by other winged helix proteins via the versatile use of the winged helix motif as a homo- or heterodimerization scaffold. PMID- 11929538 TI - Inactivation of the srtA gene in Listeria monocytogenes inhibits anchoring of surface proteins and affects virulence. AB - During infection of their hosts, Gram-positive bacteria express surface proteins that serve multiple biological functions. Surface proteins harbouring a C terminal sorting signal with an LPXTG motif are covalently linked to the cell wall peptidoglycan by a transamidase named sortase. Two genes encoding putative sortases, termed srtA and srtB, were identified in the genome of the intracellular pathogenic bacterium Listeria monocytogenes. Inactivation of srtA abolishes anchoring of the invasion protein InlA to the bacterial surface. It also prevents the proper sorting of several other peptidoglycan-associated LPXTG proteins. Three were identified by a mass spectrometry approach. The DeltasrtA mutant strain is defective in entering epithelial cells, similar to a DeltainlA mutant. In contrast to a DeltainlA mutant, the DeltasrtA mutant is impaired for colonization of the liver and spleen after oral inoculation in mice. Thus, L. monocytogenes srtA is required for the cell wall anchoring of InlA and, presumably, for the anchoring of other LPXTG-containing proteins that are involved in listerial infections. PMID- 11929539 TI - Resveratrol acts as a natural profungicide and induces self-intoxication by a specific laccase. AB - The grapevine (Vitis) secondary metabolite resveratrol is considered a phytoalexin, which protects the plant from Botrytis cinerea infection. Laccase activity displayed by the fungus is assumed to detoxify resveratrol and to facilitate colonization of grape. We initiated a functional molecular genetic analysis of B. cinerea laccases by characterizing laccase genes and evaluating the phenotype of targeted gene replacement mutants. Two different laccase genes from B. cinerea were characterized, Bclcc1 and Bclcc2. Only Bclcc2 was strongly expressed in liquid cultures in the presence of either resveratrol or tannins. This suggested that Bclcc2, but not Bclcc1, plays an active role in the oxidation of both resveratrol and tannins. Gene replacement mutants in the Bclcc1 and Bclcc2 gene were made to perform a functional analysis. Only Bclcc2 replacement mutants were incapable of converting both resveratrol and tannins. When grown on resveratrol, both the wild type and the Bclcc1 replacement mutant showed inhibited growth, whereas Bclcc2 replacement mutants were unaffected. Thus, contrary to the current theory, BcLCC2 does not detoxify resveratrol but, rather, converts it into compounds that are more toxic for the fungus itself. The Bclcc2 gene was expressed during infection of B. cinerea on a resveratrol-producing host plant, but Bclcc2 replacement mutants were as virulent as the wild-type strain on various hosts. The activation of a plant secondary metabolite by a pathogen introduces a new dimension to plant-pathogen interactions and the phytoalexin concept. PMID- 11929540 TI - Salmonella enterica serotype Typhimurium ShdA is an outer membrane fibronectin binding protein that is expressed in the intestine. AB - The shdA gene is the only determinant known to be required for persistence of Salmonella enterica serotype Typhimurium (S. Typhimurium) in the murine caecum and for efficient and prolonged shedding of the organism with the faeces. To study the biological activity of the ShdA protein, we examined its expression and binding activity. ShdA was not detected with anti-ShdA antiserum in S. Typhimurium strain ATCC14028 grown in vitro, suggesting that this protein is not expressed under standard conditions of bacterial cultivation in the laboratory. However, in mice infected with S. Typhimurium, an immunofluorescence signal detected with anti-ShdA antiserum co-localized with that generated by anti-O4 antiserum in thin sections from the caecum. Expression of the cloned shdA gene from the T7 promoter in vitro resulted in detection of ShdA in the outer membrane of S. Typhimurium and in binding of fibronectin to the bacterial surface. Binding of purified glutathione-S-transferase (GST)-ShdA fusion protein to fibronectin was dose dependent and could be partially inhibited by preincubation with antifibronectin antibodies. GST-ShdA bound to connective tissue and the basement membrane in thin sections from the murine caecum in situ. A similar labelling pattern was produced when thin sections of the murine caecum were stained with antifibronectin antiserum. Collectively, these data demonstrate that ShdA is a surface-localized, fibronectin-binding protein whose expression is induced in vivo in the murine caecum, a tissue in which a cognate receptor of this outer membrane protein is expressed. PMID- 11929541 TI - Transcytosis of iota-toxin across polarized CaCo-2 cells. AB - Iota-toxin from Clostridium perfringens type E is a binary toxin consisting of two independent proteins, an enzymatic Ia and binding Ib component. Ia catalyses ADP-ribosylation of actin monomers, thus disrupting the actin cytoskeleton. In this report, we show that Ia plus Ib applied apically or basolaterally induce a rapid decrease in the transepithelial resistance (TER) of CaCo-2 cell monolayers and disorganization of actin filaments as well as the tight and adherens junctions. Ib alone, on the apical or basolateral side, slowly decreased the TER without affecting the actin cytoskeleton, possibly via pore formation. Interestingly, the two iota-toxin components inoculated separately on each cell surface induced cytopathic effects and a TER decrease. Anti-Ib sera, raised against the whole molecule or the Ia docking domain and applied to the opposite cell side versus Ib, neutralized the TER decrease. In addition, radioactive Ib incubated in the basolateral compartment was detected on the apical side by selective cell surface biotinylation. This argues for a transcytotic routing of Ib to mediate internalization of Ia from the opposite cell surface. Bafilomycin A1 also prevented the cytopathic effects of Ia and Ib applied separately to each cell side, possibly by blocking translocation of Ia into the cytosol and/or the intracellular transport of Ib. Ib is either routed into the cell independently of Ia, trans-cytosed and permanently exposed on the opposite cell surface or continuously recycled between an endosomal compartment and the cell surface. PMID- 11929542 TI - The polyphosphate kinase plays a negative role in the control of antibiotic production in Streptomyces lividans. AB - The polyphosphate kinase gene (ppk) from Streptomyces lividans, which encodes a 774-amino-acid protein (86.4 kDa) showing extensive homology to other bacterial polyphosphate kinases, was cloned by polymerase chain reaction (PCR) using oligonucleotides derived from the putative ppk gene from the closely related species, Streptomyces coelicolor. In vitro, the purified Ppk was shown to be able to synthesize the polyphosphate [poly(P)] from ATP (forward reaction) as well as to regenerate ATP from the poly(P) in the presence of an excess of ADP (reverse reaction). In conditions of poly(P) synthesis, a phosphoenzyme intermediate was detected, indicating an autophosphorylation of the enzyme in the presence of ATP. The ppk gene was shown to be transcribed as a monocistronic mRNA from a unique promoter. Its transcription was only detectable during the late stages of growth in liquid minimal medium. A mutant strain interrupted for ppk was characterized by increased production of the antibiotic actinorhodin on rich R2YE solid medium (0.37 mM KH2PO4 added). This production was enhanced on the same medium with no KH2PO4 added but was completely abolished by the addition of 1.48 mM KH2PO4. In the ppk mutant strain, this increased production correlated with enhanced transcription of actII-ORF4 encoding the specific activator of the actinorhodin pathway. In that strain, the transcription of redD and cdaR, encoding the specific activators of the undecylprodigiosin and calcium-dependent antibiotic biosynthetic pathways, respectively, was also increased but to a lesser extent. The enhanced expression of these regulators did not seem to be related to increased relA-dependent ppGpp synthesis, as no obvious increase in relA expression was observed in the ppk mutant strain. These results suggested that the negative regulatory effect exerted by Ppk on antibiotic biosynthesis was most probably caused by the repression exerted by the endogenous Pi, resulting from the hydrolysis of the poly(P) synthesized by Ppk, on the expression of the specific activators of the antibiotic biosynthetic pathways. PMID- 11929543 TI - Identification of a gene (lpt-3) required for the addition of phosphoethanolamine to the lipopolysaccharide inner core of Neisseria meningitidis and its role in mediating susceptibility to bactericidal killing and opsonophagocytosis. AB - We have identified a gene, lpt-3, that is required for the addition of phosphoethanolamine to the 3-position (PEtn-3) on the beta-chain heptose (HepII) of the inner core lipopolysaccharide (LPS) of Neisseria meningitidis (Nm). The presence of this PEtn-3 substituent is characteristic of the LPS of a majority ( approximately 70%) of hypervirulent Nm strains, irrespective of capsular serogroup, and is required for the binding of a previously described monoclonal antibody (mAb B5) to a surface-accessible epitope. All strains of Nm that have PEtn-3 possess the lpt-3 gene. In some lpt-3-containing strains, the 3-position on HepII is preferentially substituted by glucose instead of PEtn, the result of lgtG phase variation mediated by slippage of a homopolymeric tract of cytidines. Inactivation of lpt-3 resulted in loss of PEtn-3, lack of reactivity with mAb B5 and conferred relative resistance to bactericidal killing and opsonophagocytosis by mAb B5 in vitro. Thus, the identification of lpt-3 has facilitated rigorous genetic, structural and immunobiological definition of an immunodominant epitope that is a candidate immunogen for inclusion in an LPS-based vaccine to protect against invasive meningococcal disease. PMID- 11929544 TI - Plasticity of a transcriptional regulation network among alpha-proteobacteria is supported by the identification of CtrA targets in Brucella abortus. AB - CtrA is a master response regulator found in many alpha-proteobacteria. In Caulobacter crescentus and Sinorhizobium meliloti, this regulator is essential for viability and is transcriptionally autoregulated. In C. crescentus, it is required for the regulation of multiple cell cycle events, such as DNA methylation, DNA replication, flagella and pili biogenesis and septation. Here, we report the characterization of the ctrA gene homologue in the alpha2 proteobacteria Brucella abortus, a facultative intracellular pathogen responsible for brucellosis. We detected CtrA expression in the main Brucella species, and its overproduction led to a phenotype typical of cell division defect, consistent with its expected role. A purified B. abortus CtrA recombinant protein (His6 CtrA) was shown to protect the B. abortus ctrA promoter from DNase I digestion, suggesting transcriptional autoregulation, and this protection was enhanced under CtrA phosphorylation on a conserved Asp residue. Despite the similarities shared by B. abortus and C. crescentus ctrA, the pathway downstream from CtrA may be distinct, at least partially, in both bacteria. Indeed, beside ctrA itself, only one (the ccrM gene) out of four B. abortus homologues of known C. crescentus CtrA targets is bound in vitro by phosphorylated B. abortus CtrA. Moreover, further footprinting experiments support the hypothesis that, in B. abortus, CtrA might directly regulate the expression of the rpoD, pleC, minC and ftsE homologues. Taken together, these results suggest that, in B. abortus and C. crescentus, similar cellular processes are regulated by CtrA through the control of distinct target genes. The plasticity of the regulation network involving CtrA in these two bacteria may be related to their distinct lifestyles. PMID- 11929545 TI - c-Src/Lyn kinases activate Helicobacter pylori CagA through tyrosine phosphorylation of the EPIYA motifs. AB - The human pathogen Helicobacter pylori colonizes the mucous layer of the stomach. During parasitic infection, freely swimming bacteria adhere to the gastric epithelial cells and trigger intracellular signalling pathways. This process requires the translocation of the effector protein CagA into the host cell through a specialized type IV secretion system encoded in the cag pathogenicity island. Following transfer, CagA is phosphorylated on tyrosine residues by a host cell kinase. Here, we describe how the tyrosine phosphorylation of CagA is restricted to a previously identified repeated sequence called D1. This sequence is located in the C-terminal half of the protein and contains the five-amino-acid motif EPIYA, which is amplified by duplications in a large fraction of clinical isolates. Tyrosine phosphorylation of CagA is essential for the activation process that leads to dramatic changes in the morphology of cells growing in culture. In addition, we observed that two members of the src kinases family, c Src and Lyn, account for most of the CagA-specific kinase activity in host cell lysates. Thus, CagA translocation followed by tyrosine phosphorylation at the EPIYA motifs promotes a growth factor-like response with intense cytoskeletal rearrangements, cell elongation effects and increased cellular motility. PMID- 11929546 TI - Role of thioredoxins in the response of Saccharomyces cerevisiae to oxidative stress induced by hydroperoxides. AB - Glutaredoxins and thioredoxins are highly conserved, small, heat-stable oxidoreductases. The yeast Saccharomyces cerevisiae contains two gene pairs encoding cytoplasmic glutaredoxins (GRX1, GRX2) and thioredoxins (TRX1, TRX2), and we have used multiple mutants to determine their roles in mediating resistance to oxidative stress caused by hydroperoxides. Our data indicate that TRX2 plays the predominant role, as mutants lacking TRX2 are hypersensitive, and mutants containing TRX2 are resistant to these oxidants. However, the requirement for TRX2 is only apparent during stationary phase growth, and we present three lines of evidence that the thioredoxin isoenzymes actually have redundant activities as antioxidants. First, the trx1 and trx2 mutants show wild-type resistance to hydroperoxide during exponential phase growth; secondly, overexpression of either TRX1 or TRX2 leads to increased resistance to hydroperoxides; and, thirdly, both Trx1 and Trx2 are equally able to act as cofactors for the thioredoxin peroxidase, Tsa1. The antioxidant activity of thioredoxins is required for both the survival of yeast cells as well as protection against oxidative stress during stationary phase growth, and correlates with an increase in the expression of both TRX1 and TRX2. We show that the requirement for thioredoxins during this growth phase is dependent on their activity as cofactors for the antioxidant enzyme Tsa1, and for regulation of the redox state and protein-bound levels of the low-molecular-weight antioxidant glutathione. PMID- 11929547 TI - Behaviour of topological marker proteins targeted to the Tat protein transport pathway. AB - The Escherichia coli Tat system mediates Sec-independent export of protein precursors bearing twin arginine signal peptides. Formate dehydrogenase-N is a three-subunit membrane-bound enzyme, in which localization of the FdnG subunit to the membrane is Tat dependent. FdnG was found in the periplasmic fraction of a mutant lacking the membrane anchor subunit FdnI, confirming that FdnG is located at the periplasmic face of the cytoplasmic membrane. However, the phenotypes of gene fusions between fdnG and the subcellular reporter genes phoA (encoding alkaline phosphatase) or lacZ (encoding beta-galactosidase) were the opposite of those expected for analogous fusions targeted to the Sec translocase. PhoA fusion experiments have previously been used to argue that the peripheral membrane DmsAB subunits of the Tat-dependent enzyme dimethyl sulphoxide reductase are located at the cytoplasmic face of the inner membrane. Biochemical data are presented that instead show DmsAB to be at the periplasmic side of the membrane. The behaviour of reporter proteins targeted to the Tat system was analysed in more detail. These data suggest that the Tat and Sec pathways differ in their ability to transport heterologous passenger proteins. They also suggest that caution should be observed when using subcellular reporter fusions to determine the topological organization of Tat-dependent membrane protein complexes. PMID- 11929548 TI - Saccharomyces cerevisiae Ccr4-not complex contributes to the control of Msn2p dependent transcription by the Ras/cAMP pathway. AB - The Ccr4-Not complex is a global regulator of transcription that affects genes positively and negatively and is thought to modulate the activity of TFIID. In the present work, we provide evidence that the Ccr4-Not complex may contribute to transcriptional regulation by the Ras/cAMP pathway. Several observations support this model. First, Msn2/4p-dependent transcription, which is known to be under negative control of cAMP-dependent protein kinase (PKA), is derepressed in all ccr4-not mutants. This phenotype is paralleled by specific post-translational modification defects of Msn2p in ccr4-not mutants relative to wild-type cells. Secondly, mutations in various NOT genes result in a synthetic temperature sensitive growth defect when combined with mutations that compromise cells for PKA activity and at least partially suppress the effects of both a dominant active RAS2Val-19 allele and loss of Rim15p. Thirdly, Not3p and Not5p, which are modified and subsequently degraded by stress signals that also lead to increased Msn2/4p-dependent activity, show a specific two-hybrid interaction with Tpk2p. Together, our results suggest that the Ccr4-Not complex may function as an effector of the Ras/cAMP pathway that contributes to repress basal, stress- and starvation-induced transcription by Msn2/4p. PMID- 11929549 TI - Antitermination by GlpP, catabolite repression via CcpA and inducer exclusion triggered by P-GlpK dephosphorylation control Bacillus subtilis glpFK expression. AB - The Bacillus subtilis glpFK operon encoding the glycerol transport facilitator (GlpF) and glycerol kinase (GlpK) is induced by glycerol-3-P and repressed by rapidly metabolizable sugars. Carbon catabolite repression (CCR) of glpFK is partly mediated via a catabolite response element cre preceding glpFK. This operator site is recognized by the catabolite control protein A (CcpA) in complex with one of its co-repressors, P-Ser-HPr or P-Ser-Crh. HPr is a component of the phosphoenolpyruvate:sugar phosphotransferase system (PTS), and Crh is an HPr homologue. The hprK-encoded HPr kinase phosphorylates HPr and Crh at Ser-46. But in neither ccpA nor hprK mutants was expression of a glpF'-lacZ fusion relieved from CCR, as a second, CcpA-independent CCR mechanism implying the terminator tglpFK, whose formation is prevented by the glycerol-3-P-activated antiterminator GlpP, is operative. Deletion of tglpFK led to elevated expression of the glpF' lacZ fusion and to partial relief from CCR. CCR completely disappeared in DeltatglpFK mutants carrying a disruption of ccpA or hprK. The tglpFK-requiring CCR mechanism seems to be based on insufficient synthesis of glycerol-3-P, as CCR of glpFK was absent in ccpA mutants growing on glycerol-3-P or synthesizing H230R mutant GlpK. In cells growing on glycerol, glucose prevents the phosphorylation of GlpK by P-His-HPr. P-GlpK is much more active than GlpK, and the absence of P~GlpK formation in DeltaptsHI strains prevents glycerol metabolism. As a consequence, only small amounts of glycerol-3-P will be formed in glycerol and glucose-exposed cells (inducer exclusion). The uptake of glycerol-3-P via GlpT provides high concentrations of this metabolite in the ccpA mutant and allows the expression of the glpF'-lacZ fusion even when glucose is present. Similarly, despite the presence of glucose, large amounts of glycerol-3-P are formed in a glycerol-exposed strain synthesizing GlpKH230R, as this mutant GlpK is as active as P-GlpK. PMID- 11929550 TI - Consequences of RNase E scarcity in Escherichia coli. AB - The endoribonuclease RNase E plays an important role in RNA processing and degradation in Escherichia coli. The construction of an E. coli strain in which the cellular concentration of RNase E can be precisely controlled has made it possible to examine and quantify the effect of RNase E scarcity on RNA decay, gene regulation and cell growth. These studies show that RNase E participates in a step in the degradation of its RNA substrates that is partially or fully rate determining. Our data also indicate that E. coli growth requires a cellular RNase E concentration at least 10-20% of normal and that the feedback mechanism that limits overproduction of RNase E is also able to increase its synthesis when its concentration drops below normal. The magnitude of the in-crease in RNA longevity under conditions of RNase E scarcity may be limited by an alternative pathway for RNA degradation. Additional experiments show that RNase E is a stable protein in E. coli. No other E. coli gene product, when either mutated or cloned on a multicopy plasmid, seems to be capable of compensating for an inadequate supply of this essential protein. PMID- 11929551 TI - Influences of orthodontic appliances on oral populations of mutans streptococci. AB - A study was conducted to assess the influence of fixed appliances on the mutans streptococcal in a group of patients receiving orthodontic treatment. Mutans streptococcal counts in stimulated saliva of 27 patients were obtained at baseline, and at 1-month intervals for 4 months. The difference in mutans streptococcal counts at baseline and at the 4-month sampling was less than an order of magnitude in 18 of the patients, between 1 and 2 orders of magnitude in eight of the patients, and 3 orders of magnitude in one patient. Restriction endonuclease digests of genomic DNA from representative mutans streptococci isolates taken from baseline and 4-month saliva samples, as well as from 4-month tooth and appliance surfaces, were examined by pulsed field gel electrophoresis, after restriction endonuclease digestion. Results of the DNA banding patterns associated with isolates from 19 patients showed that, for 12 patients, all isolates examined represented the same clone of Streptococcus mutans, whereas for six patients two different S. mutans clones were detected. One patient yielded three different clones of S. mutans. A much larger number of baseline, as well as post-appliance, isolates will have to be examined from each patient in future studies, in order to determine if the number of different S. mutans clones harbored by individual patients is related to orthodontic treatment. PMID- 11929552 TI - Nitric oxide production by a murine macrophage cell line (RAW264.7) stimulated with lipopolysaccharide from Actinobacillus actinomycetemcomitans. AB - The aim of this study was to determine whether Actinobacillus actinomycetemcomitans lipopolysaccharide (LPS-A. actinomycetemcomitans) could stimulate a murine macrophage cell line (RAW264.7 cells) to produce nitric oxide (NO). The cells were treated with LPS-A. actinomycetemcomitans or Escherichia coli LPS (LPS-Ec) for 24 h. The effects of N(G)-monomethyl-L-arginine (NMMA), polymyxin B and cytokines (IFN-gamma, TNF-alpha, IL-4 and IL-12) on the production of NO were also determined. The role of protein tyrosine kinase, protein kinase C and microtubulin organization on NO production were assessed by incubating RAW264.7 cells with genistein, bisindolylmaleide and colchicine prior to LPS-A. actinomycetemcomitans stimulation, respectively. NO levels from the culture supernatants were determined by the Griess reaction. The results showed that LPS-A. actinomycetemcomitans stimulated NO production by RAW264.7 cells in a dose-dependent manner, but was slightly less potent than LPS-Ec. NMMA and polymyxin B blocked the production of NO. IFN-gamma and IL-12 potentiated but IL 4 depressed NO production by LPS-A. actinomycetemcomitans-stimulated RAW264.7 cells. TNF-alpha had no effects on NO production. Genistein and bisindolylmalemaide, but not colchicine, reduced the production of NO in a dose dependent mechanism. The results of the present study suggest that A. actinomycetemcomitans LPS, via the activation of protein tyrosine kinase and protein kinase C and the regulatory control of cytokines, stimulates NO production by murine macrophages. PMID- 11929554 TI - High prevalence of cfxA beta-lactamase in aminopenicillin-resistant Prevotella strains isolated from periodontal pockets. AB - This prospective study was designed to investigate amoxicillin-resistant oral anaerobes, and to identify their beta-lactamase-encoding genes. Three subgingival bacterial samples were collected from 12 patients suffering from periodontitis. One to seven beta-lactamase-producing strains were obtained from each patient, mostly belonging to the Prevotella genus (Bacteroides eggerthii, 2/35 strains; Prevotella sp., 33/35 strains). PCR assays were used to detect cfxA and cepA/cblA, the genes encoding class A/group2e beta-lactamases previously described in the Bacteroides fragilis group. The present investigation confirmed the role of Prevotella species as beta-lactamase producers in periodontal pockets. Additionally, this PCR screening showed (1): the high prevalence of CfxA beta-lactamase production by aminopenicillin-resistant Prevotella (32/33: 97.0% positive strains) vs. cepA/cblA (1/33: 3.0% positive strains), and (2) the presence of cfxA in the periodontal reservoir in the absence of antimicrobial therapy during the previous 6 months. PMID- 11929553 TI - Oral yeast carriage in patients with advanced cancer. AB - The aim of this study was to investigate oral yeast carriage amongst patients with advanced cancer. Oral rinse samples were obtained from 120 subjects. Yeasts were isolated using Sabouraud's dextrose agar and CHROMagar Candida, and were identified using a combination of the API 20 C AUX yeast identification system, species-specific PCR and 26S rDNA gene sequencing. Oral yeast carriage was present in 66% of subjects. The frequency of isolation of individual species was: Candida albicans, 46%; Candida glabrata, 18%; Candida dubliniensis, 5%; others, < 5%. The increasing isolation of non-Candida albicans species is clinically important, since these species are often more resistant to antifungal drugs. Oral yeast carriage was associated with denture wearing (P = 0.006), and low stimulated whole salivary flow rate (P = 0.009). Identification of these risk factors offers new strategies for the prevention of oral candidosis in this group of patients. PMID- 11929555 TI - Identification of Candida dubliniensis among oral yeast isolates from an Italian population of human immunodeficiency virus-infected (HIV+) subjects. AB - Candida dubliniensis, an emerging oral pathogen, phenotypically resembles Candida albicans so closely that it is easily misidentified as such. The aim of the present study was to evaluate the usefulness of two phenotypic methods, growth at 45 degrees C and 2,3,5-triphenyltetrazolium chloride (TTC) reduction, for confirming presumptive identification of C. dubliniensis and C. albicans by colony color on CHROMagar Candida (CAC) medium. A combination of these methods was used to establish the prevalence of oral C. dubliniensis in an Italian population of 45 human immunodeficiency virus (HIV)-infected subjects. Twenty-two samples (48.9%) were positive for yeasts on CAC medium producing a total of 37 fungal isolates. The colony color and 45 degrees C growth ability test correctly identified all C. dubliniensis and C. albicans isolates (5/37, 13.5%, and 16/37, 43.2%, respectively), while assessment of TTC reduction misidentified one C. albicans isolate. The isolation rate of C. dubliniensis was 11.1% (5/45 patients). All of the C. dubliniensis isolates were highly susceptible to fluconazole (MIC = 0.5 microg/ml). The combination of CAC medium screening with growth at 45 degrees C and TTC reduction tests may represent a simple, reliable and inexpensive identification protocol for C. dubliniensis. PMID- 11929556 TI - Cariogenic traits in xylitol-resistant and xylitol-sensitive mutans streptococci. AB - Long-term xylitol consumption leads to the emergence of xylitol-resistant (X-R) mutans streptococci. The aim of the present study was to compare cariogenic traits in X-R and xylitol-sensitive (X-S) strains. Six strains of mutans streptococci, three X-R and three X-S strains, were studied. Xylitol resistance and sensitivity were confirmed by growth in xylitol-supplemented media. Acid production from glucose or fructose or uptake of xylitol was initiated by adding (14)C-labelled glucose, fructose or xylitol to bacterial suspensions. The resultant metabolites were identified by HPLC. Lactate was the major metabolite from glucose, whether the bacteria were grown in the presence or the absence of xylitol. Lactate production per colony-forming unit was lower in X-S cells than in X-R cells. Fructose was metabolized by both X-R and X-S cells. Both X-R and X S cells took up xylitol, but xylitol-5-P was detected in X-S cells only. Total polysaccharides were measured through production of C(14)-labelled ethanol insoluble polymers from [U(14)-C]-sucrose. No difference in polysaccharide production was found between X-R and X-S cells. The present study thus does not support the contention that X-R are less cariogenic than X-S mutans streptococci. PMID- 11929557 TI - Inhibitory effects of whole and parotid saliva on immunomodulators. AB - Based on the presence of cytokines in whole saliva and their association with resistance and susceptibility to infectious disease, the present study was designed to evaluate the diagnostic potential of a large panel of cytokines and chemokines in saliva. Despite the endogenous presence of Th1/Th2 and pro inflammatory cytokines and several chemokines in whole and parotid saliva of most individuals tested, the detection of known concentrations of several recombinant cytokines and chemokines was inhibited immediately following their addition to each type of saliva. In contrast, purified immunoglobulins were unaffected by either whole or parotid saliva. Further studies revealed that the inhibition of immunoreactivity involved sequestration of the majority of cytokines affected and degradation of chemokines. These results suggest that absolute concentrations of cytokines/chemokines may not be fully detectable in saliva. Therefore, the diagnostic value of any cytokine/chemokine is questionable and should be evaluated independently as such. PMID- 11929558 TI - Genetic exchange between Treponema denticola and Streptococcus gordonii in biofilms. AB - Some gram-positive oral bacteria such as Streptococcus gordonii are naturally transformable. We investigated the possibility of genetic exchange among oral bacteria using an erythromycin-resistant (Erm(r)) shuttle plasmid, pKMR4PE, as an indicator. S. gordonii Challis cells were co-cultivated with purified pKMR4PE or with Trepomena denticola harboring pKMR4PE either in broth or in artificial biofilms. Transformation of S. gordonii occurred both in broth and in biofilms with pKMR4PE or T. denticola harboring pKMR4PE as donor sources. These results provide direct experimental evidence that gene transfer can occur from T. denticola to S. gordonii. PMID- 11929559 TI - Molecular and immunological evidence of oral Treponema in the human brain and their association with Alzheimer's disease. AB - The purpose of this investigation was to use molecular and immunological techniques to determine whether oral Treponema infected the human brain. Pieces of frontal lobe cortex from 34 subjects were analyzed with species-specific PCR and monoclonal antibodies. PCR detected Treponema in 14/16 Alzheimer's disease (AD) and 4/18 non-AD donors (P < 0.001), and AD specimens had more Treponema species than controls (P < 0.001). PCR also detected Treponema in trigeminal ganglia from three AD and two control donors. Cortex from 15/16 AD subjects and 6/18 controls contained Treponema pectinovorum and/or Treponema socranskii species-specific antigens (P < 0.01). T. pectinovorum and/or T. socranskii antigens were also found in trigeminal ganglia and pons from four embalmed cadavers, and 2/4 cadavers also had Treponema in the hippocampus. These findings suggest that oral Treponema may infect the brain via branches of the trigeminal nerve. PMID- 11929560 TI - Fluoride and organic weak acids as respiration inhibitors for oral streptococci in acidified environments. AB - Oxygen metabolism (respiration) of Streptococcus mutans GS-5 involving NADH oxidases, mainly of the H(2)O-producing type, was found to be acid sensitive, as was NADH oxidase activity of cell extracts. Respiration of intact cells in acidified media was also highly sensitive to fluoride, with a 50% inhibitory concentration of about 0.02 mM at pH 4. In contrast, NADH oxidases in cell extracts were fluoride insensitive. Fluoride inhibition of respiration of intact cells was related to weak-acid effects leading to enhanced proton permeability of cells, cytoplasmic acidification and resultant acid inhibition of NADH oxidases and glycolysis. Organic weak acids, such as indomethacin and benzoate, were also effective inhibitors. H(2)O(2) production by intact cells of Streptococcus sanguis NCTC 10904, a peroxide producer, was similarly inhibited by fluoride or organic weak acids in acidified media. Thus, weak acids act as respiratory inhibitors for oral streptococci indirectly by acidifying the cytoplasm rather than acting as direct inhibitors of NADH oxidases. PMID- 11929561 TI - Bacteroides forsythus hemagglutinin is inhibited by N-acetylneuraminyllactose. AB - Bacteroides forsythus, which has been recognized as a pathogen associated with periodontitis, produces a hemagglutinin. The hemagglutinin was localized in the envelope of B. forsythus. The hemagglutinating activity was inhibited by lactose at concentrations as low as 1 mM, and by L-arginine and L-lysine at concentrations of 100 mM. N-Acetylneuraminyllactose (NeuAc-lactose) was at least 100 times more potent an inhibitor than lactose, as it completely inhibited the hemagglutination at concentrations below 10 microM. This is similar to the Helicobacter pylori hemagglutinin. The hemagglutinin was heat-labile, and resistant to treatment with proteases such as trypsin. A specific antibody raised against one of the S-layer proteins that are major species-specific proteins had no inhibitory effect on the hemmaglutination. These results suggest that the NeuAc-lactose-sensitive adhesin of B. forsythus may play an important role in colonization in the oral cavity. PMID- 11929562 TI - Differences in Candida albicans adhesion to intact and denatured type I collagen in vitro. AB - An inhibition assay of Candida albicans adhesion to gelatin-immobilized membranes was compared with that to intact type I collagen-immobilized membranes using an arginine-glycine-aspartic acid (RGD) containing peptide. As compared with a protein-free membrane, gelatin and collagen significantly enhanced the adherence of C. albicans. The adhesion of the yeast to gelatin was significantly inhibited by the RGD peptides, but not by arginine-glycine-glutamic acid (RGE) peptides. In contrast, attachment to collagen was not inhibited by RGD peptides. These results suggest that the RGD sequence of gelatin and the integrin-like proteins of yeasts may be involved in adherence. PMID- 11929565 TI - The relationship between regional cerebral blood flow and the Wisconsin Card Sorting Test in negative schizophrenia. AB - The purpose of this study was to explore the relationship between regional cerebral blood flow (rCBF) and problem-solving thinking in negative schizophrenia. Twenty-one negative schizophrenic patients and 12 normal controls were studied with single photon emission computed tomography (SPECT). Measures of regional cerebral blood flow (rCBF) were taken both at rest and during a prefrontal activation task using Wisconsin Card Sorting Test (WCST). Compared with controls, poor performances on the WCST of total trials category (TT), perseverative errors (PE) and non-perseverative errors (NE) were found in negative schizophrenic (P < 0.05). During WCST activation, patients showed interhemispheric differences in the prefrontal region, but under rest conditions, no such differences manifested. The negative schizophrenia group had a significantly lower rCBF change rate in profrontal lobe during stimulant WCST than those in normal controls (P < 0.05). The negative schizophrenic patient has executive function deficits and lower rCBF perfusion in left profrontal lobes, which suggest that the negative schizophrenic patient has dysfunction of the left profrontal region. PMID- 11929563 TI - Bacteriology and antimicrobial susceptibility of gram-positive cocci isolated from pus specimens of orofacial odontogenic infections. AB - We recently reported the beta-lactamase production and antimicrobial susceptibility of anaerobic gram-negative rods isolated from pus specimens of 93 orofacial odontogenic infections. In this report, we determine the bacteriology and antimicrobial susceptibility of bacteria other than anaerobic gram-negative rods, mainly gram-positive cocci, isolated from the same specimens. Streptococcus constellatus and Peptostreptococcus micros were frequent isolates from all types of infection examined. Peptostreptococcus prevotii, Corynebacterium species, and Eubacterium species were recovered only from dentoalveolar infections, while Gemella morbillorum was found more frequently in periodontitis than in the other infections. beta-Lactamase-positive strains were detected only in staphylococci. Ampicillin, ampicillin/sulbactam, cefazolin, cefotaxime, imipenem, erythromycin, clindamycin and levofloxacin showed high susceptibility rates (> or = 77%) against viridans streptococci, Peptostreptococcus and Gemella. Minocycline showed a high MIC90 value against viridans streptococci (32 microg/ml), and metronidazole was effective against Peptostreptococcus and Gemella. These results provide useful information for the treatment of orofacial odontogenic infections. PMID- 11929566 TI - Body image, symptoms and insight in chronic schizophrenia. AB - Historically, schizophrenics' body image problems were regarded as related particularly to delusions and hallucinations. However, during the 1980s, the predominant view of the phenomenology of the disorder broadened to include negative symptoms; deviations in schizophrenics' body image underlie various behaviors or allegations concerning the body and should be refocused. The present study attempted to detect body image deviations in chronic schizophrenia using the Body Image Questionnaire (BIQ), which comprises three hypothetical components (anatomical, functional and other psychological components), and to clarify their related clinical characteristics in symptoms and insight. The BIQ was administered to 93 chronic schizophrenics (diagnosed according to DSM IV; 44 men and 49 women) and 177 normals (78 men and 99 women) adults. The combined data of the three BIQ components in schizophrenic and normal subjects were factor analyzed separately, and factor scores obtained were compared between schizophrenic and normal groups. The factor scores that differentiated groups were further compared between schizophrenic subgroups, determined by high or low scores for positive symptoms assessed by Scale for the Assessment of Positive Symptoms, negative symptoms by Scale for the Assessment of Negative Symptoms and insight by the Schedule for Assessing Insight. Significant differences between diagnostic groups were found in five of nine factor scores. Dullness in movement, powerlessness, unusually strong gastrointestinal function, lifelessness and fragility proved to be the deviated body images in chronic schizophrenic patients. Powerlessness and lifelessness proved to be related to positive and negative symptoms, and unusually strong gastrointestinal function and fragility to insight. PMID- 11929567 TI - Hoarding in obsessive-compulsive disorder and related disorders: a preliminary report of 15 cases. AB - Hoarding, the repetitive collection of excessive quantities of poorly useable items of little or no value with failure to discard these items over time, is characterized in DSM-IV as a symptom of obsessive-compulsive disorder (OCD) and obsessive-compulsive personality disorder (OCPD) but has, until recently, received scant empirical investigation. We describe the demographics, phenomenology, associated psychopathology and family history in 15 subjects presenting with hoarding behavior. Fifteen subjects were recruited from an OCD clinic and newspaper advertisement and assessed with the comprehensive Structured Clinical Interview for DSM-IV (SCID I and II), the Yale-Brown Obsessive Compulsive Scale (Y-BOCS), and a hoarding questionnaire (devised by the authors). The sample comprised 11 women and four men who hoarded a mean of seven item types, with a mean duration of 13.2 +/- 3.9 years (range 2-15 years). Their mean age was 41.8 +/- 14.3 years (range 20-65 years). The most common motive for hoarding was the fear of discarding items of practical value. Nine subjects met DSM-IV criteria for OCD, 9 met criteria for OCPD, for symptoms and behaviors other than hoarding, while six subjects met criteria for a putative OCD spectrum disorder (Tourette's, body dysmorphic disorder, trichotillomania). Six subjects reported little or no control over their hoarding, but only one subject saw her symptoms as an 'illness' warranting treatment. Pathological hoarding is usually a covert and chronic behavior causing distress and/or impairment, and may be related to OCD and OCPD. Hoarding may meet the criterion for a compulsion in DSM IV, yet there is evidence to suggest that hoarding may manifest in a variety of other psychiatric conditions. While a range of pharmacologic and behavioral treatments have been tried, their effectiveness in managing hoarding behaviors requires additional research. PMID- 11929568 TI - Relationship between clock drawing test performance and regional cerebral blood flow in Alzheimer's disease: a single photon emission computed tomography study. AB - The clock drawing test (CDT) is a useful tool for screening cognitive impairment. Previous neuropsychological studies have revealed that CDT performance requires several cognitive functions including semantic memory, visuospatial function and executive function. However, the neural substrates involved remain to be elucidated. The aim of the present study was to identify the brain regions responsible for CDT performance in patients with Alzheimer's disease (AD). Twenty six patients satisfying the National Institute of Neurological and Communicative Disease and Stroke-Alzheimer's Disease and Related Disorders Association criteria for probable AD underwent the CDT and the Mini-Mental State Examination (MMSE), together with a N-isopropyl-p-[123I] iodoamphetamine (IMP) single photon emission computed tomography measurements of the resting regional cerebral blood flow (rCBF). The CDT score correlated significantly with the MMSE score (r=0.582; P < 0.05). Stepwise multiple regression analysis revealed that the MMSE score and the left posterior temporal rCBF were major predictors of CDT score. These findings suggest that the CDT score may reflect the severity of dementia, and that it has a close relationship with the left posterior temporal function. Our findings provide the first functional neuroimaging evidence for the neural substrates involved in CDT performance. PMID- 11929570 TI - Psychological characteristics of eating disorders as evidenced by the combined administration of questionnaires and two projective methods: the Tree Drawing Test (Baum Test) and the Sentence Completion Test. AB - The objective of this study is to examine psychological/psychopathological characteristics of eating disorders and their subtypes through a combined administration of questionnaires and projective tests. Three questionnaires (Eating Disorder Inventory - 2, Social Adaptation Scale, Southern California University Eating Disorder Inventory - Revised) and two projective tests (the Tree Drawing Test [TDT, Baum Test], and the Sentence Completion Test [SCT]) were administered to 126 female patients between the ages of 15 and 30 years, with eating disorders according to DSM-IV criteria at our outpatient clinic, and to 54 sex- and age-matched control subjects. The purging subtypes of eating disorders (anorexia nervosa - binge-eating/purging type [ANBP] and bulimia nervosa - purging type [BNP]) were clearly differentiated from the controls, both by the questionnaires and the projective tests. Compared with the controls, ANBP/BNP showed more problematic profiles across the three questionnaires, drew smaller and poorer trees in TDT to a more left location on the drawing paper, and gave fewer positive, and more negative responses in SCT. In contrast, few significant differences were found between anorexia nervosa- restricting type (ANR) and the controls, and between ANBP and BNP. As a trend, however, ANR was consistently located between the controls and ANBP/BNP across the whole questionnaires and projective tests. PMID- 11929569 TI - Changes in regional cerebral blood flow abnormalities in late-life depression following response to electroconvulsive therapy. AB - The impact of electroconvulsive therapy (ECT) on the regional cerebral blood flow (rCBF) abnormalities in late-life depression is still unknown and the clinical significance of these findings in late-life depression has not been fully discussed. Using single photon emission computed tomography (SPECT) with 99mTc hexamethylpropylene amine oxime (99mTc-HMPAO), we examined the changes of rCBF patterns in nine late-life patients with major depressive episodes before and following response to ECT compared with nine age- and sex-matched healthy volunteers. Statistical comparisons were made on both region-of-interest (ROI) and voxel-by-voxel bases. In ROI-based analyses, a mean rCBF was significantly decreased in the patients before ECT compared with the control, significantly increased (normalized) in the patients 2 weeks after ECT compared with that before ECT, and still increased in the patients 12 weeks after ECT compared with that before ECT. In voxel-by-voxel analyses, a significant rCBF reduction was found in the bilateral pre- and subcallosal anterior cingulate cortex, the bilateral caudal orbitofrontal cortex, the right insular cortex and the right posterior middle frontal gyrus in patients before ECT compared with the control, and similar rCBF patterns were shown at 2 weeks and 12 weeks after ECT. We propose the hypothesis that a mean rCBF reduction may have a state-related property while persistent anterior paralimbic hypoperfusion may have a trait-like property, which relates to the relapse vulnerability as well as a tendency toward medication failure and illness chronicity in late-life depression. PMID- 11929571 TI - Mental health of two communities of Japanese-Brazilians: a comparative study in Japan and in Brazil. AB - The aim of the present study was to assess the mental health status of Japanese Brazilians living in Kiyoharadai, Japan and compare the findings with the Japanese-Brazilians living in Bauru, Brazil. A comparative community-based mental health survey was conducted from November 1997 to April 1999 on a randomly selected sample of Brazilians of Japanese descent living in Bauru, Brazil (n=213) and on the entire Brazilian community of Kiyoharadai, Utsunomiya, Japan (n=158), using the self-reporting questionnaire (SRQ-20). Analysis was done by chi2, Fisher and multiple logistic regression. Scores indicating probable cases of minor psychiatric disorders, were found in 3.2% of the community in Bauru city and 17.8% (OR=7.01) of the community in Kiyoharadai. The sociodemographic data indicated that those with high SRQ-20 scores were most likely to be female (OR=2.98), smokers (OR=2.76), and those whose former occupation was student when living in Brazil (OR=9.57). The mental health status of the Japanese-Brazilians living in Kiyoharadai, Japan is significantly worse than that of the community living in Bauru, Brazil, particularly among women, smokers and those who were students when living in Brazil. Further research concerning the mental health of this community is necessary and assistance provided. PMID- 11929572 TI - Cerebral blood flow changes in general paresis following penicillin treatment: a longitudinal single photon emission computed tomography study. AB - Three cases of general paresis were successfully treated with high-dose penicillin. In all cases, cerebrospinal fluid cell counts decreased to normal and mental status improved rapidly. Cerebrospinal fluid protein concentrations decreased and IQ scores and overall levels of functioning improved gradually over 1 year. Regional cerebral blood flow (CBF) changes were analyzed longitudinally for 1 year using single photon emission computed tomography (SPECT). Autoradiography method and region of interest (ROI) analyses were used for quantitative CBF assessment and 3D stereotactic surface projections (3D-SSP) analyses were used for qualitative CBF pattern assessment. 3D-SSP could not reveal remarkable CBF pattern changes through the courses. The ROI analyses showed remarkable CBF decreases in all brain regions 1 month after the treatment, which recovered to normal levels 1 year after the treatment. These results suggest that remarkable decreases of quantitative CBF counts reflect the disappearance of encephalitis, while their gradual recovery reflects the gradual improvement of cerebral functional activity. As Treponema pallidum infection affects whole brain and CBF changes globally, quantitative CBF assessment may be more efficient than qualitative CBF pattern analyses for the purpose of understanding the pathophysiology of general paresis. PMID- 11929573 TI - Prevalence of dementia in the older Japanese-Brazilian population. AB - The prevalence of dementing disorders in Campo Grande of a community of Japanese Brazilians who immigrated from Okinawa was studied. Previous reports showed that the dietary pattern in Japanese immigrants in Brazil, which characterized by a low fish and large meat intake, is possibly responsible for increased risk of cardiovascular diseases compared with Japanese in Okinawa. A total of 157 persons over 70-year-old were examined, and 19 cases were diagnosed as having dementia. The prevalence (cases/100 aged 70-year-older) was 12.1 for all types of dementia, 5.7 for Alzheimer's disease (AD), 0.6 for vascular dementia (VD), 4.5 for mixed dementia (AD/VD) and 1.3 for other types of dementia. There was no case of dementia with Lewy bodies or frontotemporal lobar degeneration. These results are similar to many previous studies in Western countries and some recent surveys in Japan, and clearly show that more AD than VD appears even in the Japanese Brazilian population. The higher prevalence rate of dementia in Japanese Brazilians compared with several studies in Japan may indicate the importance of dietary factors rather than genetic factors. PMID- 11929574 TI - Subcortical overactivation in untreated schizophrenic patients: a functional magnetic resonance image finger-tapping study. AB - Functional magnetic resonance imaging (fMRI) is a well established, non-invasive technique for mapping the working brain. Yet imaging of subcortical regions has proven to be difficult. We studied 40 subjects performing an unilateral self paced finger-tapping task. Patients with schizophrenia according to DSM-IV treated with olanzapine (n =10) or haloperidol (n=10) were compared to healthy controls (n =10) and untreated patients (n=10). Brainvoyager software was used for data-analyzing. All subjects showed highly significant activation in the contralateral sensorimotor area, the supplementary motor area and the ipsilateral cerebellum. In every investigated subject contralateral subcortical regions were also significantly activated (P < 0.001). Activation in ipsilateral pallidum was significantly higher in untreated patients compared with the other groups indicating an increase in subcortical coactivation. In addition, significant correlations were revealed within groups. This study emphasizes the possibility of investigating subcortical brain activation in patients with schizophrenia. The results of the present study outline the importance of further fMRI studies to investigate interindividual activation differences under different conditions especially focusing on basal ganglia. PMID- 11929575 TI - Static stabilometry in patients with migraine and tension-type headache during a headache-free period. AB - The vestibulospinal system was evaluated using a stabilometric method in patients with migraine and episodic tension-type headache during headache-free periods. Migraine patients often complain of dizziness or vertigo during headache attacks and some exhibit these symptoms between attacks. Computerized static stabilometry is a reliable and non-invasive technique to evaluate the equilibrium function in various diseases. The subjects consisted of 21 patients with migraine, 12 patients with episodic tension-type headache and, age- and sex-matched controls. We performed two sets of static stabilometric measurements with eyes open (EO) and eyes closed (EC) for 30 s. The averages of two sessions of the following six stabilometric parameters were used for the analysis: locus length (LNG), environmental area (ENV-AREA), rectangle area (REC-AREA), locus length per second, locus length per environ area (L/E), and root mean square area. Romberg quotients (EC/EO) of these six parameters were also analyzed. The mean values of LNG, ENV-AREA and REC-AREA in the EC session in the migraine group were significantly greater than those in the controls (P < 0.05, Mann-Whitney rank sum test). Romberg quotients of all stabilometric parameters except the L/E in the migraine group were significantly greater than in the controls. Patients with episodic tension-type headache did not show any differences in the stabilometric study from the controls. The present findings suggest that patients with migraine show a significant increase of the body sway during the EC session, which indicates an underlying dysfunction in the vestibulospinal system. PMID- 11929576 TI - Validation study of the Japanese version of the Obsessive-Compulsive Drinking Scale. AB - The Obsessive-Compulsive Drinking Scale (OCDS) is a self-rating questionnaire that measures cognitive and behavioral aspects of craving for alcohol. The OCDS consists of two subscales: the obsessive thoughts of drinking subscale (OS) and the compulsive drinking subscale (CS). This study aims to validate the Japanese version of the OCDS. First, internal consistency and discriminant validity were evaluated. Second, a prospective longitudinal 3-month outcome study of 67 patients with alcohol dependence who participated in a relapse prevention program was designed to assess the concurrent and predictive validity of the OCDS. The OCDS demonstrated high internal consistency. The OS had high discriminant validity, while the CS did not. Twenty-three patients (34.3%) dropped out of treatment. These patients had significantly higher OS scores than those who completed the program. At 3 months, the relapse group had significantly higher OCDS scores than the no relapse group. Also, the OCDS score was higher in subjects who had early-onset alcohol dependence than late-onset dependence. The OCDS is useful for evaluating cognitive aspect of craving and predicts dropout and relapse. PMID- 11929577 TI - The influence on the schizophrenic symptoms by the DRD2Ser/Cys311 and -141C Ins/Del polymorphisms. AB - The hyperactivity of dopaminergic systems is one of the major etiological hypotheses of schizophrenia. The major support for this hypothesis is that effective antipsychotic drugs bind to dopamine receptors and improve acute schizophrenic symptoms. For this reason, we investigated the allelic association between schizophrenia and polymorphisms of the DRD2 genes for the Ser/Cys311 and 141C Ins/Del. The subjects were 190 schizophrenics (120 males and 70 females) and 103 normal controls (53 males and 50 females). There were no significant differences between the patients and controls in the allele frequencies and the frequencies of the genotypes. We found no statistical association between schizophrenia and polymorphisms of the DRD2 genes for the Ser/Cys311 and -141C Ins/Del. These results indicate that the DRD2 gene may not develop schizophrenia. Next, we examined whether the genotypes influence the symptoms of schizophrenia the using Positive and Negative Symptom Scale scores. The Ser/Cys patients exhibited significantly lower positive and negative symptom scores than Ser/Ser patients. Patients with Del/Del, Ins/Del, or Ins/Ins showed higher positive symptom scores in descending order. This result suggested that the Del allele worsens the positive symptoms. We concluded that the DRD2 receptor gene may not influence the onset of schizophrenia, but there is a strong possibility that the Cys311 and -141C Del have a significant influence on the symptoms of schizophrenia. PMID- 11929578 TI - The characteristics of side-effects of bromperidol in schizophrenic patients. AB - The characteristics of the side-effects of bromperidol was investigated in 33 acutely exacerbated schizophrenic patients. The most frequently observed side effects were extrapyramidal symptoms. Acute dystonia developed in 10 of 33 patients, and the mean age was significantly lower (P < 0.05) in patients with dystonia (27.3 +/- 6.2 years) than that in patients without dystonia (41.5 +/- 12.9 years). Plasma drug concentrations were not associated with side-effects. These findings suggest that acute dystonia is affected by age factor, and that daily dosage or monitoring of drug concentration is unlikely to be a useful marker for the prediction of side-effects during bromperidol treatment. PMID- 11929579 TI - Risperidone failed to improve polydipsia-hyponatremia of the schizophrenic patients. AB - The effect of risperidone on polydipsia-hyponatremia was evaluated in six hospitalized schizophrenic patients. The normalized diurnal weight gain (NDWG), urine-specific gravity (USG), urine and plasma osmolarity, and serum sodium were monitored during 9 months of risperidone treatment. The dose of risperidone (mean +/- SD=8.0 +/- 1.0, range=6-9 mg/day) was determined as approximately half of the haloperidol-equivalent dose of previous neuroleptics. Before risperidone treatment, the mean (+/- SD) BPRS score was 23.5 +/- 7.1; no significant improvement was observed after risperidone (22.0 +/- 7.5). The subjects showed relatively high serum prolactin before risperidone treatment (mean +/- SD=16.5 +/ 9.7 ng/mL), that was not significantly decreased by risperidone (14.2 +/- 7.9 ng/mL). The monthly means (+/- SD) of NDWG and USG before risperidone were 5.5 +/ 1.5 (%) and 1.002 +/- 0.001, respectively. These and other indices did not significantly improve throughout the study period. Although the sample size is relatively small, our preliminary data showed that risperidone might not be effective on polydipsia-hyponatremia of schizophrenic patients. PMID- 11929580 TI - On the role of quantitative brain imaging in the differential diagnosis of speech disorders. AB - We present the case of a 71-year-old woman with an 11-year history of slowly progressive decline of motor speech. Normal clinical investigations including routine magnetic resonance imaging (MRI) at 7, 8 and 10 years after the onset of speech dysfunction led to the suggestion of a psychogenic disorder. Extensive clinical, neuropsychological and neuroimaging investigations including 18F desoxyglucose-positron emission tomography (PET), quantitative MRI and MR spectroscopy were performed to look for subtle brain pathology. Quantitative assessment of 3D-MRI, F-desoxyglucose-PET and magnetic resonance spectroscopy all demonstrated clear evidence of multifocal frontotemporal brain pathology that had not been picked up on routine MRI investigations on previous admissions. This is the longest benign history of slowly progressive anarthria reported so far. It demonstrates a possible role of quantitative neuroimaging techniques in the diagnosis of complex neuropsychiatric disorders. PMID- 11929581 TI - A case of diffuse neurofibrillary tangles with calcification. AB - We report a 79-year-old female with atypical senile dementia with Fahr-type calcification. The patient started to show memory disturbance at the age of 75 years, followed by visual hallucination, stereotypy, personality changes such as irritability, aggression and disinhibition. Brain computed tomography (CT) demonstrated bilateral and symmetric calcification of the basal ganglia and thalamus. Magnetic resonance imaging (MRI) revealed diffuse cortical atrophy pronounced in the fronto-temporal areas. On MRI T1-weighted images the calcified areas showed a mixture of low- and high-intensity signals. Based on the overlapping clinical symptoms of Alzheimer's disease and Pick's disease, together with the brain CT and MRI findings, we clinically diagnosed the patient as having 'diffuse neurofibrillary tangles with calcification' (DNTC). The characteristics of psychiatric symptoms and neuroradiological findings in DNTC are discussed. PMID- 11929586 TI - The quantity of naturally processed peptides stably bound by HLA-A*0201 is significantly reduced in the absence of tapasin. AB - Tapasin plays a critical role in promoting peptide binding by major histocompatibility complex (MHC) class I molecules in the endoplasmic reticulum. In its absence, cell surface expression of most allotypes is significantly reduced. Two exceptions are HLA-A*0201 and HLA-B*2705. In this study, the repertoire of peptides bound endogenously by these allotypes in the absence of tapasin was examined and stability of the HLA class I/peptide complexes assessed. Similar quantities of peptides were recovered from B*2705 complexes expressed in the absence and presence of tapasin and the composition of the peptide pools were not radically different. However, the stability of B*2705 molecules expressed at the surface of tapasin-deficient cells was found to be reduced which suggests there are subtle changes to the peptide repertoire. The impact of the absence of tapasin was more dramatic for A*0201. Although equivalent levels of cell surface A*0201 are expressed in the presence and absence of tapasin, very little A*0201 glycoprotein was recovered from tapasin-deficient cells suggesting the complexes readily dissociate. Consistent with reduced stability, A*0201 complexes were found to be rapidly lost from the surface of tapasin-deficient cells. Analysis of the small quantity of endogenously bound peptides recovered from A*0201 expressed in the absence of tapasin revealed a complex mixture typical of A*0201 molecules expressed in normal cells. Therefore these molecules are unable to exploit the alternative supply of TAP-independent A*0201-binding peptides present in the endoplasmic reticulum. Loading of A*0201 with peptides from both TAP-dependent and TAP-independent sources is significantly compromised without tapasin. PMID- 11929585 TI - Large sharing of T-cell epitopes and natural ligands between HLA-B27 subtypes (B*2702 and B*2705) associated with spondyloarthritis. AB - HLA-B*2702 is an ankylosing spondylitis-associated allotype that differs from the more common B*2705 at residues 77, 80, and 81, in the peptide-binding site. The diversity and fine specificity of alloreactive cytolytic T-lymphocyte (CTL) raised against B*2702 were analyzed at the clonal level. Significant crossreaction with B*2705 and B*2709 indicated that the three subtypes share numerous T-cell epitopes. However, some epitopes shared by B*2702 and B*2705 were lost in B*2709, which correlates with weaker association of this subtype to disease. Clonal specificities were donor-dependent, indicating that allo immunogenicity is variable among individuals. Anti-B*2702 CTL were little affected by single mutations mimicking B*2702/B*2705 polymorphism, but the double mutant at positions 77 and 81 was recognized worse than B*2705, suggesting a compensatory effect of residue 80. Thus, HLA-B27 polymorphism modulated alloreactivity through cooperative and compensatory effects on T-cell epitope structure. Comparison of B*2705- and B*2702-bound peptide repertoires revealed that they overlapped by 73% and 81%, respectively. This was larger than B*2702/B*2705 cross-reaction, indicating that HLA-B27 allospecificity is only partially determined by the nature of peptide repertoires. The large sharing of natural ligands and T-cell epitopes is consistent with a pathogenetic role of B*2702 and B*2705 in spondyloarthritis based on antigen presentation. PMID- 11929587 TI - Polymorphisms in the tumor necrosis factor (TNF) genes are associated with susceptibility to effects of ultraviolet-B radiation on induction of contact hypersensitivity. AB - We investigated the allelic distributions of single nucleotide polymorphisms (SNPs) of the TNFA, TNFB and IKBL genes, 3 microsatellites within the tumor necrosis factor (TNF) region of HLA locus, and the HLA phenotypes as well as the TLR4 gene in Chromosome 9 in 26 healthy Caucasian volunteers. These individuals were also assessed as ultraviolet B (UVB)-susceptible (S) or UVB-resistant (R). Our results identified 12 UVB-S and 14 UVB-R individuals. Attempts to correlate particular HLA-A, -B, -C, and -DR antigens with the UVB phenotypes failed. Similarly, attempts to correlate SNP at the NcoI-RFLP within intron 1 of the TNFB, IKBL and TLR4 gene with UVB phenotypes also failed. However, microsatellite analyses of TNFa, TNFc, and TNFd markers revealed a significant increase in the frequencies of TNFa2 in UVB-S individuals (P=0.00032) and of TNFd3 in UVB-R individuals (P=0.012). Moreover, DNA sequencing analyses of 5 SNPs of the TNFA promoter region revealed a significant increase in the frequency of the allele B of the TNFA gene (TNFApB) representing the nucleotide A at position -863 and C at position -1031 (P=0.015). Since it is known that TNFa2 and TNFApB is a high TNF alpha responder, whereas TNFd3 is a TNF-alpha low responder, we propose that the TNF region of HLA contains polymorphic genes that confer susceptibility and resistance to the deleterious effects of UVB radiation on the induction of contact hypersensitivity. This proposal is consistent with previous reports that a unique microsatellite region of the Tnfa gene in mice contains alleles that dictate the UVB-dependent phenotypes in mice, and implicate TNF-alpha as the primary mediator of the immune-damaging effects of UVB radiation. PMID- 11929588 TI - Inflammatory bowel disease is associated with a novel promoter polymorphism of natural resistance-associated macrophage protein 1 (NRAMP1) gene. AB - Inflammatory bowel diseases (IBD) representing both Crohn's disease and ulcerative colitis are characterized by chronic activation of macrophages. Natural resistance-associated macrophage protein 1 (NRAMP1) gene regulates macrophage activation of antimicrobial activity and has many pleiotropic effects on macrophage functions. To explore the role of the NRAMP1 gene in IBD susceptibility, we examined the promoter sequence of the NRAMP1 gene whose polymorphisms have been reported to influence the transcriptional activity of the NRAMP1 gene. One novel allele (allele 7) and two previously reported alleles (alleles 2 and 3) have been determined in a Japanese population. We investigated the association of IBD with these three alleles. The allele frequency of allele 7 was significantly higher in patients with Crohn's disease (11.1%) and ulcerative colitis (11.2%) than those in the healthy control group (4.5%) (Pc=0.015, Pc=0.018, respectively). Therefore, our findings suggest that the novel promoter polymorphism of the NRAMP1 gene may influence susceptibility to IBD in the Japanese population. PMID- 11929589 TI - The distribution of HLA class II susceptible/protective haplotypes could partially explain the low incidence of type 1 diabetes in continental Italy (Lazio region). AB - HLA class II is the primary susceptibility gene to type 1 diabetes and the analysis of HLA class II association could help to clarify the relative weight of genetic contribution to the incidence of the disease. Here we present an extensive typing for HLA class II alleles and their haplotypes in a homogenous population of type 1 diabetic patients (n=134) and controls (n=128) and in simplex (n=100) and multiplex families (n=50) from continental Italy (Lazio region). Among the various haplotypes tested, the DRB1*0301-DQA1*0501-DQB1*0201 was the most frequent found in type 1 diabetic patients and was transmitted in 82% of affected siblings, whereas DRB1*0402-DQA1*0301-DQB1*0302 appeared to have the highest odds ratio (10.4), this haplotype was transmitted in 96.3% of affected siblings, followed by DRB1*0405-DQA1*0301-DQB1*0302, DRB1*0405-DQA1*0301 DQB1*0201, DRB1*0401-DQA1*0301-DQB1*0302 and DRB1*0404-DQA1*0301-DQB1*0302. The following haplotypes showed a significant decreased transmission to diabetic siblings: DRB1*0701-DQA1*0201-DQB1*0303, DR2-DQA1*01-DQB1*0602, DR5-DQA1*0501 DQB1*0301. We suggest that the HLA DR/DQ haplotype/genotype frequencies observed could in part explain the low incidence of type 1 diabetes registered in Lazio region (8.1/100.000/year), for a number of reasons: i) the low frequency, in the general control population, of the most susceptible haplotypes and genotype for type 1 diabetes DRB1*0301-DQA1*0501-DQB1*0201 (14%), and DR4-DQA1*0301-DQB1*0302 (9%) and DRB1*0301-DQA1*0501-DQB1*0201/DR4-DQA1*0301-DQB1*0302 (0.8%) compared to other countries characterised by high incidence rate of the disease, Sardinia and Finland, respectively; ii) a significant lower ratio, in the control population, between the susceptible DRB1*0301-DQA1*0501-DQB1*0201 and the neutral DRB1*0701 DQA1*0501-DQB1*0201 haplotypes compared to the Sardinian population; iii) the high frequency of protection haplotypes/genotypes as the DR5-DQA1*0501-DQB1*0301, and DR5-DQA1*0501-DQB1*0301/DR5-DQA1*0501-DQB1*0301 very common in the control population of Lazio region and the DRB1*1401-DQA1*0101-DQB1*0503 haplotype. PMID- 11929590 TI - Combination of HLA-A and HLA class II alleles controls the susceptibility to rheumatoid arthritis. AB - Two hundred and four unrelated Japanese patients with rheumatoid arthritis (RA) were typed for HLA by serological typing and DNA typing. The serological typing revealed that frequencies of HLA-A11, DR4, DR53 and DQ4 were increased and those of DR8 and DQ1 were decreased in the patients. The DNA typing has precisely defined the disease-associated HLA class II alleles; DRBl*0405, DQAl*03 and DQBl*0401 showed positive associations, while negative associations were found with DRBl*0803, DQAl*0103 and DQBl*0601. The risk for RA was found to be closely associated with particular amino acid sequences of DR-beta chain, including glycine residue at the 86th position in addition to those between 70 and 74, which are known to confer binding specificity and affinity to antigenic peptides. The observation that the frequency of HLA-A11 was increased in the DRBl*0405 positive patients suggested the interaction of these two alleles in the susceptibility to RA. On the other hand, the frequency of DPB1*0201 was increased in the DRBl*0405-negative patients and the frequency of HLA-A2 was increased in the DPBl*0201-positive patients, especially in the younger onset group. These findings suggested that the combination of HLA-A2 and DPBl*0201 may confer the susceptibility in the DRBl*0405-negative patients. Our results suggested the possibility that the susceptibility to RA is controlled by the interaction of HLA A and DRBl genes or by that of HLA-A and DPBl genes in different patient subgroups. PMID- 11929591 TI - The distribution of DQ genes in the Saharawi population provides only a partial explanation for the high celiac disease prevalence. AB - Celiac disease (CD) is a multifactorial disorder of the small intestine caused by a permanent dietary intolerance to gluten. The combined presence of the HLA class II DQA1*0501 and DQB1*0201 alleles represents the major genetic component for disease predisposition. It has been shown that the Saharawi refugees living in northern Africa have a very high frequency of CD. In the present study we analysed this population to evaluate the degree of association with CD of the haplotypes and genotypes at the main HLA-DQB1 and DQA1 disease loci. We found a strong association of the DR3, DQB1*0201-DQA1*0501-positive haplotypes and genotypes. A very high frequency of DR3, DQB1*0201-DQA1*0501 was also observed in the general Saharawi population. These results indicate that there is a good correlation between disease prevalence and frequency of the main predisposing haplotype in the background population. However, the correlation is incomplete because similar frequencies of DR3 are also observed in populations such as the Sardinians showing a much lower prevalence of CD. We can conclude that the distribution of DQ genes in the Saharawi population only provides a partial explanation for the high prevalence of CD. Other factors, such as rapidly changing dietary habits and/or non-DQ genes, may also play some role. PMID- 11929592 TI - Strong association of a tumor necrosis factor-alpha promoter allele with cerebral malaria in Myanmar. AB - To investigate the host genetic factors affecting the clinical course of falciparum malaria, polymorphism of the tumor necrosis factor-alpha (TNF-alpha) promoter region was analyzed in patients with cerebral malaria. Two hundred and forty-three Myanmar patients with falciparum malaria at Mae Sot Malaria clinic and Mae Sot General Hospital located at the border between Thailand and Myanmar, were included in this study. Among the patients (128 from Karen, 115 from Burma), 200 were uncomplicated and 43 had cerebral malaria. The TNF-alpha 5'- flanking region showed biallelic polymorphic sites at -238, -308, -857, -863, -1031, and there were 7 alleles (TNFP-A, B, C, D, M1, M4, M7) found in the patients from Myanmar. We found that the TNFP-D allele was significantly associated with cerebral malaria in the populations from Karen (Pc<0.0001, OR=124.86) and Burma (Pc<0.0001, OR=34.50). TNFP-D showed no significant linkage disequilibrium with any alleles of HLA-B or HLA-DRB1, suggesting that TNFP-D was primarily associated with cerebral malaria in Myanmar. PMID- 11929593 TI - Linkage disequilibria between HLA-B, C1_4_1, MICA and MICB. AB - The polymorphisms of MICA exon 5 (5 alleles), MICB intron 1 (13 alleles), C1_4_1 (6 alleles), HLA-B (29 alleles) and HLA-A (15 alleles) were investigated in a healthy German population. Sequencing was performed for the MICB alleles CA14, CA15, CA17, CA23 and CA26 isolated from different cell lines. Variation to the published sequence was observed for CA14, CA15 and for CA17. At the C1_4_1 locus a new allele (CAAA)9 was identified and confirmed by sequencing. Linkage disequilibria were investigated for two-point- and three-point-haplotypes. Although the average relative delta value correlates loosely with the physical distance from HLA-B to MICB: HLA-B-C1_4_1>HLA-B-MICA>HLA-B-MICB, there are several exceptions to this rule. Analyzing three-point-haplotypes for the segment MICB to HLA-A a wide variation of linkage disequilibria for some of the classical HLA-A, B haplotypes has been observed. While the HLA-A1, B8 haplotype displays strong relative delta values over the entire distance from HLA-A to MICB, other haplotypes have linkage disequilibria only in a limited region. PMID- 11929594 TI - The nucleotide diversity of MICA and MICB suggests the effect of overdominant selection. AB - The major histocompatibility complex (MHC) class I-related molecules A and B (MICA and MICB) are stress-inducible cell surface antigens that are recognized by immunocytes bearing the receptor NKG2D. In our study we estimated the average number of synonymous (pis) and nonsynonymous nucleotide substitutions (pia) per site in exons 2-4 of MICA and MICB. In exons 2 and 3 of MICB only nonsynonymous substitutions were found, and in exon 3 of MICA pia clearly exceeded pis. This finding is in contrast to the evolutionary parameters found in most other genes, and is reminiscent of the elevated pia values caused by overdominant selection in the peptide-binding region of conventional MHC class I molecules. It may be explained by the hypothetical interaction with nonpeptide antigens, or by resistance against pathogens. PMID- 11929595 TI - Characterization of a novel O(1) variant allele at the ABO blood group locus. AB - We describe the identification and molecular characterization of a novel variant O(1) allele of the ABO blood group locus. The allele was found in a young child and by analyzing the maternal DNA we were able to show that a meiotic recombination event between the maternal O(1v-3) and B(1-1) alleles recreated a O(1)/B hybrid allele. Further characterization of intron 6 sequences delineated the putative recombination breakpoint between nucleotide position 42 and 163 of the intron. We propose that the novel O(1variant) allele should be named O(1v-7) and is a combination of exon 6 from a O(1v-3) allele and exon 7 from a B(1-1) allele. PMID- 11929596 TI - CD antigens 2001. AB - The most recent Human Leucocyte Differentiation Antigen Workshop ("HLDA7") took place in 2000 in Harrogate, UK and the proceedings are about to be published (Leucocyte Typing VII). New Sections were introduced in this Workship (Dendritic cells, Stem/progenitor cells, Erythroid cells and Carbohydrate Structures) and monoclonal antibodies were selected for which at least some molecular data were already available (to avoid "blind" screening of reagents against known specificities). A total of more than 80 new CD specificities were established (previously the average was less than 30 new CD specificities per Workshop) and these are listed in this article. There is already evidence for the existence of many new leucocyte surface molecules for study at the next HLDA Workshop (in Adelaide in 2004), and we have listed in this article a number of such potential CD candidates (identified following the production of monoclonal antibodies or via gene cloning). There are also today an increasing number of lineage- and/or stage-restricted leucocyte-associated molecules localised within the cell cytoplasm (or nucleus): they will certainly prove of intense in the future for many laboratories studying human haematopoietic cells (regardless of whether a new "intracellular CD" categorisation scheme is devised for such molecules). PMID- 11929599 TI - Junctions as organizing centers in epithelial cells? A fly perspective. AB - Junctions isolate the apical from the basal-lateral surfaces and are required for the polarization of epithelial cells. Genetics of junction formation in Drosophila has identified numerous transmembrane and scaffolding proteins that assemble in membrane-associated complexes. This review focuses on recent findings suggesting that junctions are autonomous organizing centers for the establishment and maintenance of apical-basal polarity in epithelial cells as well as other polarized processes such as the orientation of the division apparatus. PMID- 11929598 TI - Sorting out signals in fly endosomes. AB - Ligands and receptors that mediate cell-cell interactions during development are removed from the cell surface by endocytosis. Subsequently, many of these internalized proteins are detected in multivesicular bodies (MVBs). Recent work in different organisms has elucidated some aspects of MVB biogenesis and trafficking. This review discusses some intriguing links between these findings, the sorting of proteins in endocytic trafficking, and the regulation of signaling pathways in Drosophila. PMID- 11929600 TI - Morphogen gradient formation and vesicular trafficking. AB - Morphogens are secreted signaling molecules which form spatial concentration gradients while moving away from a restricted source of production. A simple model of gradient formation postulates that the morphogens dilute as they diffuse between cells. In this review we discuss recent data supporting the idea that movement of the morphogen could also occur via vesicular trafficking through the cells. We explore the implications of these results for the control of gradient formation and the determination of the gradient slope which ultimately encodes the coordinates of positional information. PMID- 11929601 TI - Ubiquitin-independent entry into the yeast recycling pathway. AB - The yeast a-factor receptor (Ste3p) is subject to two mechanistically distinct modes of endocytosis: a constitutive, ligand-independent pathway links to vacuolar degradation of the receptor, while a ligand-dependent uptake pathway links primarily to recycling and thus, receptor reutilization. Ste3p ubiquitination triggers its uptake into the constitutive pathway. The present work considers the role of the receptor ubiquitination associated with the Ste3p ligand-dependent endocytosis mechanism. The doa4delta mutation which reduces the cellular availability of ubiquitin blocks the Ste3p constitutive uptake. Uptake into the Ste3p ligand-dependent recycling pathway, however, continues unimpaired. The ubiquitin independence of Ste3p ligand-dependent uptake was further indicated by analysis of receptor mutants having Lys-to-Arg substitutions at all possible ubiquitin acceptor sites. Again, the ligand-induced internalization was unimpaired. Furthermore, no discernible effect was seen on either recycling or on the slow PEP4-dependent turnover of the receptor (for receptor internalized via the ligand-dependent mechanism, trafficking to the vacuole/lysosome is the minor, alternate fate to recycling). However, one striking effect of the Lys-to-Arg mutations was noted. Following a prolonged exposure of the cells to the a-factor ligand, rather than being delivered to the vacuolar lumen, the Lys-to-Arg receptor was found to localize instead to the limiting membrane of the vacuole. Thus, while receptor ubiquitination clearly is not required for either the a factor-dependent uptake into recycling pathway or for the recycling itself, it does affect the routing of receptor to the vacuole, likely by affecting the routing through the late endosomal, multivesicular body: ubiquitinated receptor may be selected into the internal, lumenal vesicles, while unmodified receptor may be left to reside at the limiting external membrane. PMID- 11929602 TI - Melanophilin, the product of the leaden locus, is required for targeting of myosin-Va to melanosomes. AB - The formation of complex subcellular organelles requires the coordinated targeting of multiple components. Melanosome biogenesis in mouse melanocytes is an excellent model system for studying the coordinated function of multiple gene products in intracellular trafficking. To begin to order events in melanosome biogenesis and distribution, we employed the classical coat-color mutants ashen, dilute, and leaden, which affect melanosome distribution, but not melanin synthesis. The loci have been renamed Rab27a, Myo5a, and Mlph for their gene products. While each of the three loci has been shown to be required for melanosome distribution, the point(s) at which each acts is unknown. We have utilized primary melanocytes to examine the interdependencies between rab27a, myosin-Va, and melanophilin. The localization of rab27a to melanosomes did not require the function of either myosin-Va or melanophilin, but leaden function was required for the association of myosin-Va with melanosomes. In leaden melanocytes permeabilized before fixation, myosin-Va immunoreactivity was greatly attenuated, suggesting that myosin-Va is free in the cytoplasm. Finally, we have complemented both the leaden and ashen phenotypes by cell fusion and observed redistribution of mature melanosomes in the absence of both protein and melanin synthesis. Together, our data suggest a model for the initial assembly of the machinery required for melanosome distribution. PMID- 11929603 TI - Transient surface delivery of invariant chain-MHC II complexes via endosomes: a quantitative study. AB - Newly synthesized major histocompatibility complex class II needs to be directed to late endocytic compartments to combine with peptide antigens. Efficient transport requires complexes of major histocompatibility complex class II and invariant chain (alphabetaIi). Since such complexes have been detected on the plasma membrane in human cells, this compartment was proposed as the primary destination for alphabetaIi exiting the trans-Golgi network. Here, I have used density gradient electrophoresis and selective biotinylation to investigate the trafficking route of alphabetaIi quantitatively. Density gradient electrophoresis analysis showed that alphabetaIi was transported from the trans-Golgi network to endosomes at approximately 1.7% min-1. Surface delivery of alphabetaIi was delayed relative to endosome transport by approximately 10 min and showed slower kinetics ( approximately 0.4% min-1), suggesting that alphabetaIi reached the plasma membrane only after arrival in endosomes. A biotinylation assay revealed that 20-40% of endosomal alphabetaIi was delivered to the plasma membrane at steady state, suggesting that surface alphabetaIi was entirely derived from endosomes. Surface alphabetaIi was rapidly re-internalized and either returned to the cell surface or accessed degradative compartments. Peptide loading commenced approximately 30 min after delivery to endosomes. Thus alphabetaIi directly traffics from trans-Golgi network to endosomes and enters an endosome-plasma membrane 'carousel' until transport to peptide-loading compartments ensues. PMID- 11929604 TI - An alternate targeting pathway for procathepsin L in mouse fibroblasts. AB - In transformed mouse fibroblasts, a significant proportion of the lysosomal cysteine protease cathepsin L remains in cells as an inactive precursor which associates with membranes by a mannose phosphate-independent interaction. When microsomes prepared from these cells were resolved on sucrose gradients, this procathepsin L was localized in dense vesicles distinct from those enriched for growth hormone, which is secreted constitutively when expressed in fibroblasts. Ultrastructural studies using antibodies directed against the propeptide to avoid detection of the mature enzyme in lysosomes revealed that the proenzyme was concentrated in dense cores within small vesicles and multivesicular endosomes which labeled with antibodies specific for CD63. Consistent with the resemblance of these cores to those of regulated secretory granules, secretion of procathepsin L from fibroblasts was modestly stimulated by phorbol, 12-myristate, 13-acetate. When protein synthesis was blocked with cycloheximide and lysosomal proteolysis inhibited with leupeptin, procathepsin L was found to gradually convert to the active single-chain protease. The data suggest that when synthesis levels are high, a portion of the procathepsin L is packaged in dense cores within multivesicular endosomes localized near the plasma membrane. Gradual activation of this proenzyme achieves targeting of the proenzyme to lysosomes by a mannose phosphate receptor-independent pathway. PMID- 11929605 TI - The dark side of lysosome-related organelles: specialization of the endocytic pathway for melanosome biogenesis. AB - Melanosomes are lysosome-related organelles within which melanin pigments are synthesized and stored in melanocytes and retinal pigment epithelial cells. Early ultrastructural studies of pigment cells revealed that melanosomes consist of a complex series of organelles; more recently, these structures have been correlated with cargo constituents. By studying the fate of melanosomal and endosomal cargo in melanocytic cells, the effects of disease-related mutations on melanosomal morphology, and the genes affected by these mutations, we are beginning to gain novel insights into the biogenesis of these complex organelles and their relationship to the endocytic pathway. These insights demonstrate how specialized cells integrate unique and ubiquitous molecular mechanisms in subverting the endosomal system to generate cell-type specific structures and their associated functions. Further dissection of the melanosomal system will likely shed light not only on the biogenesis of lysosome-related organelles but also on general aspects of vesicular transport in the endosomal system. PMID- 11929606 TI - Kinetic timing: a novel mechanism that improves the accuracy of GTPase timers in endosome fusion and other biological processes. AB - The GTPase superfamily contains a large number of proteins that function as molecular switches by binding and hydrolyzing GTP molecules. They are localized at various intracellular organelles and control diverse cellular processes. For many GTPases, the lifetime of the activated, GTP-bound state is believed to serve as a timer in determining the activation time of a biological event such as membrane fusion and signal transduction. However, such a timer is intrinsically stochastic due to thermal noise at the level of single GTPase molecules. Here, we describe a mathematical model that shows how a directional GTPase cycle, in a nonequilibrium steady-state driven by GTP hydrolysis, can significantly reduce the variance in the lifetime of an activated GTPase molecule and thereby increase the accuracy and efficiency of the timer. This mechanism, termed kinetic timing, articulates a clear function for the energy consumption in GTPase-controlled biological processes. It provides a rationale for why biological timers utilize a GTP hydrolysis cycle rather than a simple GTP binding-dissociation equilibrium, and why the GTP-bound state is a better timer than the GDP-bound state. It also explains the necessity for the existence of multiple GTP-bound intermediates identified by fluorescence spectroscopy and nuclear magnetic resonance studies. PMID- 11929607 TI - The protein coat in membrane fusion: lessons from fission. AB - Multiple cell biological processes involve two opposite rearrangements of membrane configuration, referred to as fusion and fission. While membrane intermediates in protein-mediated fusion have been studied in some detail, the global force which drives sequential stages of the fusion reaction from early local intermediates to an expanding fusion pore remains unknown. Fusion proceeds via stages, which are analogous but in the opposite direction to that of membrane budding-off and fission driven by protein coats. On the basis of this analogy, we propose that an interconnected coat formed by membrane-bound activated fusion proteins surrounding the membrane contact zone generates the driving force for fusion. This fusion protein coat has a strongly curved intrinsic shape opposite to that of the protein coat driving fission. To relieve internal stresses, the fusion protein coat spontaneously bends out of the initial shape of the membrane surface. This bending produces elastic stresses in the underlying lipid bilayer and drives its fusion with the apposing membrane. The hypothesis that 'bystander' proteins (i.e. fusion proteins outside the contact zone) generate the driving force for fusion offers a new interpretation for a number of known features of the fusion reaction mediated by the prototype fusion protein, influenza hemagglutinin, and might bring new insights into mechanisms of other fusion reactions. PMID- 11929608 TI - Apo AI/ABCA1-dependent and HDL3-mediated lipid efflux from compositionally distinct cholesterol-based microdomains. AB - We have investigated whether a raft heterogeneity exists in human monocyte derived macrophages and fibroblasts and whether these microdomains are modulated by lipid efflux. Triton X-100 (Triton) or Lubrol WX (Lubrol) detergent-resistant membranes from cholesterol-loaded monocytes were associated with the following findings: (i) Lubrol-DRM contained most of the cellular cholesterol and at least 75% of Triton-detergent-resistant membranes. (ii) 'Lubrol rafts', defined by their solubility in Triton but insolubility in Lubrol, were enriched in unsaturated phosphatidylcholine and showed a lower cholesterol to choline phospholipid ratio compared to Triton rafts. (iii) CD14 and CD55 were recovered in Triton- and Lubrol-detergent-resistant membranes, whereas CD11b was found exclusively in Triton DRM. ABCA1 implicated in apo AI-mediated lipid efflux and CDC42 were partially localized in Lubrol- but not in Triton-detergent-resistant membranes. (iv) Apo AI preferentially depleted cholesterol and choline phospholipids from Lubrol rafts, whereas HDL3 additionally decreased the cholesterol content of Triton rafts. In fibroblasts, neither ABCA1 nor CDC42 was found in Lubrol rafts, and both apo AI and HDL3 reduced the lipid content in Lubrol- as well as in Triton-detergent-resistant membranes. In summary, we provide evidence for the existence of compositionally distinct membrane microdomains in human cells and their modulation by apo AI/ABCA1-dependent and HDL3-mediated lipid efflux. PMID- 11929609 TI - Selective delivery of secretory cargo in Golgi-derived carriers of nonepithelial cells. AB - In epithelial cells, soluble cargo proteins destined for basolateral or apical secretion are packaged into distinct trans-Golgi network-derived transport carriers. Similar carriers, termed basolateral- and apical-like, have been observed in nonepithelial cells using ectopically expressed membrane marker proteins. Whether these cells are capable of selectively packaging secretory proteins into distinct carriers is still an open question. Here, we have addressed this issue by analyzing the packaging and transport of secretory human chromogranin B fusion proteins using a green fluorescent protein-based high resolution, dual-color imaging technique. We were able to show that these secretory markers were selectively packaged at the Golgi into tubular/vesicular like transport carriers containing basolateral membrane markers, resulting in extensive cotransport. In contrast, deletion mutants of the human chromogranin B fusion proteins lacking an N-terminal loop structure were efficiently transported in both basolateral- and apical-like carriers, the latter displaying a spherical morphology. Similarly, in polarized epithelial cells, the human chromogranin B fusion protein was secreted basolaterally and the loop-deleted analogue into both the basolateral and apical medium. These findings suggest that nonepithelial cells, like their epithelial counterparts, possess a sorting machinery capable of selective packaging of secretory cargo into distinct types of carriers. PMID- 11929610 TI - Characterization of novel Rab6-interacting proteins involved in endosome-to-TGN transport. AB - Rab6 GTPase regulates intracellular transport at the level of the Golgi complex. Using the yeast two-hybrid screen, we have isolated two clones that specifically interact with the three isoforms of Rab6 present in mammalian cells (Rab6A, A' and B). The cDNAs encode two proteins of 976 and 1120 amino acids (calculated molecular mass of 112 and 128 kDa, respectively) that we named Rab6IP2A and Rab6IP2B (for Rab6 Interacting Protein 2). The two proteins likely correspond to spliced variants of the same gene. Rab6IP2s have no significant homology with other known proteins, including Rab effectors or partners. They are ubiquitously expressed, mostly cytosolic and found in high molecular mass complexes in brain cytosol. We show that Rab6IP2s can be recruited on Golgi membranes in a Rab6:GTP dependent manner. The overexpression of any form of Rab6IP2 has no detectable effect on the secretory pathway. In contrast, the retrograde transport of the Shiga toxin B subunit between the plasma membrane and the Golgi complex is partly inhibited in cells overexpressing the Rab6-binding domain of Rab6IP2. Our data suggest that Rab6IP2s is involved in the pathway regulated by Rab6A'. PMID- 11929611 TI - Calcium-dependent transferrin receptor recycling in bovine chromaffin cells. AB - The release of regulated secretory granules is known to be calcium dependent. To examine the Ca2+-dependence of other exocytic fusion events, transferrin recycling in bovine chromaffin cells was examined. Internalised 125I-transferrin was released constitutively from cells with a half-time of about 7 min. Secretagogues that triggered catecholamine secretion doubled the rate of 125I transferrin release, the time courses of the two triggered secretory responses being similar. The triggered 125I-transferrin release came from recycling endosomes rather than from sorting endosomes or a triggered secretory vesicle pool. Triggered 125I-transferrin release, like catecholamine secretion from the same cells, was calcium dependent but the affinities for calcium were very different. The extracellular calcium concentrations that gave rise to half maximal evoked secretion were 0.1 mm for 125I-transferrin and 1.0 mm for catecholamine, and the intracellular concentrations were 0.1 microm and 1 microm, respectively. There was significant 125I-transferrin recycling in the virtual absence of intracellular Ca2+, but the rate increased when Ca2+ was raised above 1 nm, and peaked at 1 microm when the rate had doubled. Botulinum toxin type D blocked both transferrin recycling and catecholamine secretion. These results indicate that a major component of the vesicular transport required for the constitutive recycling of transferrin in quiescent cells is calcium dependent and thus under physiological control, and also that some of the molecular machinery involved in transferrin recycling/fusion processes is shared with that for triggered neurosecretion. PMID- 11929612 TI - RNA:(guanine-N2) methyltransferases RsmC/RsmD and their homologs revisited- bioinformatic analysis and prediction of the active site based on the uncharacterized Mj0882 protein structure. AB - BACKGROUND: Escherichia coli guanine-N2 (m2G) methyltransferases (MTases) RsmC and RsmD modify nucleosides G1207 and G966 of 16S rRNA. They possess a common MTase domain in the C-terminus and a variable region in the N-terminus. Their C terminal domain is related to the YbiN family of hypothetical MTases, but nothing is known about the structure or function of the N-terminal domain. RESULTS: Using a combination of sequence database searches and fold recognition methods it has been demonstrated that the N-termini of RsmC and RsmD are related to each other and that they represent a "degenerated" version of the C-terminal MTase domain. Novel members of the YbiN family from Archaea and Eukaryota were also indentified. It is inferred that YbiN and both domains of RsmC and RsmD are closely related to a family of putative MTases from Gram-positive bacteria and Archaea, typified by the Mj0882 protein from M. jannaschii (1dus in PDB). Based on the results of sequence analysis and structure prediction, the residues involved in cofactor binding, target recognition and catalysis were identified, and the mechanism of the guanine-N2 methyltransfer reaction was proposed. CONCLUSIONS: Using the known Mj0882 structure, a comprehensive analysis of sequence-structure-function relationships in the family of genuine and putative m2G MTases was performed. The results provide novel insight into the mechanism of m2G methylation and will serve as a platform for experimental analysis of numerous uncharacterized N-MTases. PMID- 11929613 TI - The farnesyl transferase inhibitor RPR-130401 does not alter radiation susceptibility in human tumor cells with a K-Ras mutation in spite of large changes in ploidy and lamin B distribution. AB - BACKGROUND: Growth inhibition by RPR-130401, a non-peptidomimetic farnesyltransferase inhibitor, was investigated without or with combined exposure to ionizing radiation in three human tumor cell lines (HCT-116, MiAPaCa-2 and A 549) bearing a point mutation in the K-Ras gene. RESULTS: RPR-130401 inhibited cell growth with an IC50 of 50 nM (HCT-116), 120 nM (MiAPaCa-2) and 710 nM (A 549), with a poor incidence of apoptosis. The drug brought about G1 and S phase depletion together with arrest of cells in G2 phase and induced a significant accumulation of hyperploid cells showing active S phase DNA synthesis, with HCT 116 and A-549 cells being the most and least responsive, respectively. The drug also produced dramatic changes of the nuclear lamin B pattern, without lamin B cleavage and perturbation of the actin cytoskeleton. On the other hand, RPR 130401 elicited strictly additive interaction in combined treatment with ionizing radiation with regard to cell kill, altered cell cycle progression and induced hyperploidy. CONCLUSIONS: The data suggest that disruption of orderly progression through mitosis and cytokinesis, is a major outcome of drug action and that this effect proceeds from inhibition of lamin B farnesylation. It is anticipated from the strict additivity of RPR-130401 and radiation that neither induced radiation resistance nor acute or late complications of radiotherapy, should occur in combined treatment with RPR-130401. PMID- 11929614 TI - "Bronchial artery delivery of viral vectors for gene delivery in cystic fibrosis; superior to airway delivery?". AB - BACKGROUND: Attempts at gene therapy for the pulmonary manifestations of Cystic Fibrosis have relied mainly on airway delivery. However the efficiency of gene transfer and expression in the airway epithelia has not reached therapeutic levels. Access to epithelial cells is not homogenous for a number of reasons and the submucosal glands cannot be reached via the airways. PRESENTATION: We propose to inject gene delivery vectors directly into bronchial arteries combined with pre-delivery of vascular endothelial growth factor to increase vascular endothelial permeability and post-delivery flow reduction by balloon occlusion. Thus it may be possible to reach mucous secreting cells of the bronchial luminal epithelium and the submucosal glands in an increased and homogenous fashion. TESTING: This combination of techniques to the best of our knowledge has not previously been investigated, and may enable us to overcome some of the current limitations to gene therapy for Cystic Fibrosis. PMID- 11929615 TI - Pressure-controlled versus volume-controlled ventilation: does it matter? AB - Volume-controlled ventilation (VCV) and pressure-controlled ventilation (PCV) are not different ventilatory modes, but are different control variables within a mode. Just as the debate over the optimal ventilatory mode continues, so too does the debate over the optimal control variable. VCV offers the safety of a pre-set tidal volume and minute ventilation but requires the clinician to appropriately set the inspiratory flow, flow waveform, and inspiratory time. During VCV, airway pressure increases in response to reduced compliance, increased resistance, or active exhalation and may increase the risk of ventilator-induced lung injury. PCV, by design, limits the maximum airway pressure delivered to the lung, but may result in variable tidal and minute volume. During PCV the clinician should titrate the inspiratory pressure to the measured tidal volume, but the inspiratory flow and flow waveform are determined by the ventilator as it attempts to maintain a square inspiratory pressure profile. Most studies comparing the effects of VCV and PCV were not well controlled or designed and offer little to our understanding of when and how to use each control variable. Any benefit associated with PCV with respect to ventilatory variables and gas exchange probably results from the associated decelerating-flow waveform available during VCV on many ventilators. Further, the beneficial characteristics of both VCV and PCV may be combined in so-called dual-control modes, which are volume-targeted, pressure-limited, and time-cycled. PCV offers no advantage over VCV in patients who are not breathing spontaneously, especially when decelerating flow is available during VCV. PCV may offer lower work of breathing and improved comfort for patients with increased and variable respiratory demand. PMID- 11929616 TI - Closed-loop mechanical ventilation. AB - Closed-loop mechanical ventilation encompasses a plethora of techniques, ranging from the very simple to the relatively complex. In the simplest form, closed-loop ventilation is the control of one output variable of the mechanical ventilator based on the measurement of an input variable. An example would be pressure support ventilation, in which flow (output) is constantly changing to maintain pressure (input) constant throughout inspiration. More complex forms of closed loop ventilation involve measurement of multiple inputs (eg, compliance, oxygen saturation, respiratory rate) to control multiple outputs (eg, ventilator frequency, airway pressure, tidal volume). The latter type of control more closely mimics the ventilatory control and response of human physiology. This review discusses both currently available closed-loop ventilation techniques and those only available outside the United States, along with some cutting-edge techniques that have only limited use. The operation, theoretical advantages, and limitations of each technique are reviewed. When available, the literature supporting or refuting each technique will be reviewed, but, unfortunately, little has been published on certain techniques. PMID- 11929617 TI - Weaning from mechanical ventilation. AB - Invasive mechanical ventilation can be lifesaving for patients with acute respiratory failure, but numerous complications have been identified. Therefore, once clinical improvement has occurred, emphasis is placed on quickly weaning (ie, liberating) the patient from mechanical ventilation. Weaning can be subdivided into 2 components: readiness testing and progressive withdrawal. Traditionally, both clinical factors and weaning predictors have been used to assess readiness for spontaneous breathing trials, which can be carried out using a T-piece or a low level of ventilatory support. The role of weaning predictors is under investigation, and their role in clinical decision making remains poorly defined. Recent insights into the pathophysiology of weaning failure have provided a framework for identifying potentially correctable limiting factors. Randomized controlled trials suggest that several approaches to progressive withdrawal may be acceptable, though only a minority of patients require progressive withdrawal. Emerging evidence indicates that protocol-directed weaning, driven by respiratory therapists and intensive care nurses, can improve outcome. PMID- 11929618 TI - The role of tracheostomy in weaning from mechanical ventilation. AB - A common clinical observation is that patients wean more rapidly from mechanical ventilation following tracheotomy. Expected changes in tube resistance and dead space are not adequate to explain this observation in adult patients. Theoretical considerations are too complicated to allow evaluation of expected changes in work of breathing following tracheotomy. The resistance of the upper airway is about the same as breathing quietly through an 8.0 mm endotracheal tube; however, many patients experience a higher work of breathing following extubation. This is not true in infants, in whom the reduction in airway diameter is profound and a marked reduction in resistance is seen following extubation. The other benefits of tracheostomy include better secretion removal, improved oral hygiene, less laryngeal damage, and ability to eat and speak. These should be considered when proposing this procedure. There may be less late ventilator-associated pneumonia following early tracheotomy. The assumed better safety of tracheostomy has been questioned. That patients appear to wean more rapidly is probably accounted for by the variety of factors mentioned above. PMID- 11929619 TI - Extubation. AB - As many as 20% of extubated patients require reintubation (ie, extubation failure) within 72 hours of extubation, with the exact prevalence depending on numerous factors. The pathophysiologic basis of extubation failure is often different from the cause of weaning failure. Extubation failure substantially prolongs the duration of mechanical ventilation, intensive care unit stay, and hospital stay, and substantially increases hospital mortality. Therefore, prediction of extubation outcome and prevention of extubation failure may be critically important. Unfortunately, standard weaning tests have not proven sufficiently accurate in predicting extubation outcome. New semi-objective measurements of cough strength and secretion volume can help identify patients at increased risk for extubation failure. It is important to note that mortality increases with reintubation delay, which indicates that clinical worsening may take place during the period without ventilatory support. Therefore, improved outcome may derive from rapid identification of patients at increased risk, followed by expeditious reinstitution of ventilatory support when extubation failure occurs. PMID- 11929620 TI - The long-term outcomes of mechanical ventilation: what are they and how should they be used? AB - In the critical care setting, usually the most important outcome is survival. However, this is not the only important outcome of critical care. There are increasing data that the patient's quality of life and functional status can be affected long after an intensive care unit stay, and some data suggest that mechanical ventilation strategies could influence those outcomes. Critical care clinicians' decisions regarding mechanical ventilation and related treatments such as level of sedation might have more profound and far-reaching residual effects than has been previously recognized. To deliver effective, cost effective, and patient-centered care, critical-care clinicians must consider outcomes other than survival. These outcomes include such diverse concepts as quality of life, functional status, and neuropsychological function. This review addresses theoretical and practical challenges to measuring and interpreting those other outcomes. PMID- 11929621 TI - Invasive mechanical ventilation in adults: conference summary. PMID- 11929622 TI - Bayesian QTL mapping using skewed Student-t distributions. AB - In most QTL mapping studies, phenotypes are assumed to follow normal distributions. Deviations from this assumption may lead to detection of false positive QTL. To improve the robustness of Bayesian QTL mapping methods, the normal distribution for residuals is replaced with a skewed Student-t distribution. The latter distribution is able to account for both heavy tails and skewness, and both components are each controlled by a single parameter. The Bayesian QTL mapping method using a skewed Student-t distribution is evaluated with simulated data sets under five different scenarios of residual error distributions and QTL effects. PMID- 11929623 TI - Non-random mating for selection with restricted rates of inbreeding and overlapping generations. AB - Minimum coancestry mating with a maximum of one offspring per mating pair (MC1) is compared with random mating schemes for populations with overlapping generations. Optimum contribution selection is used, whereby DeltaF is restricted. For schemes with DeltaF restricted to 0.25% per year, 256 animals born per year and heritability of 0.25, genetic gain increased with 18% compared with random mating. The effect of MC1 on genetic gain decreased for larger schemes and schemes with a less stringent restriction on inbreeding. Breeding schemes hardly changed when omitting the iteration on the generation interval to find an optimum distribution of parents over age-classes, which saves computer time, but inbreeding and genetic merit fluctuated more before the schemes had reached a steady-state. When bulls were progeny tested, these progeny tested bulls were selected instead of the young bulls, which led to increased generation intervals, increased selection intensity of bulls and increased genetic gain (35% compared to a scheme without progeny testing for random mating). The effect of MC1 decreased for schemes with progeny testing. MC1 mating increased genetic gain from 11-18% for overlapping and 1-4% for discrete generations, when comparing schemes with similar genetic gain and size. PMID- 11929624 TI - Comparison between estimation of breeding values and fixed effects using Bayesian and empirical BLUP estimation under selection on parents and missing pedigree information. AB - Bayesian (via Gibbs sampling) and empirical BLUP (EBLUP) estimation of fixed effects and breeding values were compared by simulation. Combinations of two simulation models (with or without effect of contemporary group (CG)), three selection schemes (random, phenotypic and BLUP selection), two levels of heritability (0.20 and 0.50) and two levels of pedigree information (0% and 15% randomly missing) were considered. Populations consisted of 450 animals spread over six discrete generations. An infinitesimal additive genetic animal model was assumed while simulating data. EBLUP and Bayesian estimates of CG effects and breeding values were, in all situations, essentially the same with respect to Spearman's rank correlation between true and estimated values. Bias and mean square error (MSE) of EBLUP and Bayesian estimates of CG effects and breeding values showed the same pattern over the range of simulated scenarios. Methods were not biased by phenotypic and BLUP selection when pedigree information was complete, albeit MSE of estimated breeding values increased for situations where CG effects were present. Estimation of breeding values by Bayesian and EBLUP was similarly affected by joint effect of phenotypic or BLUP selection and randomly missing pedigree information. For both methods, bias and MSE of estimated breeding values and CG effects substantially increased across generations. PMID- 11929625 TI - Multiple trait model combining random regressions for daily feed intake with single measured performance traits of growing pigs. AB - A random regression model for daily feed intake and a conventional multiple trait animal model for the four traits average daily gain on test (ADG), feed conversion ratio (FCR), carcass lean content and meat quality index were combined to analyse data from 1449 castrated male Large White pigs performance tested in two French central testing stations in 1997. Group housed pigs fed ad libitum with electronic feed dispensers were tested from 35 to 100 kg live body weight. A quadratic polynomial in days on test was used as a regression function for weekly means of daily feed intake and to describe its residual variance. The same fixed (batch) and random (additive genetic, pen and individual permanent environmental) effects were used for regression coefficients of feed intake and single measured traits. Variance components were estimated by means of a Bayesian analysis using Gibbs sampling. Four Gibbs chains were run for 550000 rounds each, from which 50000 rounds were discarded from the burn-in period. Estimates of posterior means of covariance matrices were calculated from the remaining two million samples. Low heritabilities of linear and quadratic regression coefficients and their unfavourable genetic correlations with other performance traits reveal that altering the shape of the feed intake curve by direct or indirect selection is difficult. PMID- 11929626 TI - Food resource allocation patterns in lactating females in a long-term selection experiment for litter size in mice. AB - Resource allocation patterns, as quantified by residual food intake (RFI), and the consequences for offspring development were investigated during lactation in 96 females of a mouse line selected for 104 generations for high litter size at birth (S-line) and in 87 females of a non-selected control line (C-line). Litters of 45 C-line dams (Cs) and 48 S-line dams (Ss) were standardised (s) at birth; other dams (ns) supported total number of pups born (Cns and Sns, respectively). RFI during lactation was significantly lower in Sns-dams than in C-line dams and Sns-dams. After weaning Sns-dams seemed to be able to restore the negative resource situation. Sns-pups were about 25% less mature than Cns-pups at all times. Maturity was similar for Cs- and Ss-pups from 2 d in lactation on, and about 18% and 53% higher than Cns- and Sns-pups. The pre-weaning mortality rate was significantly higher in Sns-litters (35.6 +/-2.76) than in Cns-litters (4.95 +/-2.23). The results suggest that S-line dams allocated considerably more resources to maintenance of offspring than C-line dams. This was insufficient to provide the offspring with an adequate amount of resources, resulting in reduced pup development and increased pre-weaning mortality rates. PMID- 11929627 TI - Association of a missense mutation in the bovine leptin gene with carcass fat content and leptin mRNA levels. AB - Previously, we have shown that alleles of the BM1500 microsatellite, located 3.6 kb downstream of the leptin gene in cattle, were associated with carcass fat measures in a population of 154 unrelated beef bulls. Subsequently, a cytosine (C) to thymine (T) transition that encoded an amino acid change of an arginine to a cysteine was identified in exon 2 of the leptin gene. A PCR-RFLP was designed and allele frequencies in four beef breeds were correlated with levels of carcass fat. The T allele was associated with fatter carcasses and the C allele with leaner carcasses. The frequencies of the SNP alleles among breeds indicated that British breeds have a higher frequency of the T allele whereas the continental breeds have a higher occurrence of the C allele. A ribonuclease protection assay was developed to quantify leptin mRNA in a separate group of animals selected by genotype. Animals homozygous for thymine expressed higher levels of leptin mRNA. This may suggest that the T allele, which adds an extra cysteine to the protein, imparts a partial loss of biological function and hence could be the causative mutation. PMID- 11929629 TI - SRY-related genes in the genome of the rice field eel (Monopterus albus). AB - The mammalian sex determining gene, SRY, is the founding member of the new growing family of Sox (SRY-like HMG-box gene) genes. Sox genes encode transcription factors with diverse roles in development, and a few of them are involved in sex determination and differentiation. We report here the existence of Sox genes in the rice field eel, Monopterus albus, and DNA sequence information of the HMG box region of five Sox genes. The Sox1, Sox4 and Sox14 genes do not have introns in the HMG box region. The Sox9 gene and Sox17 gene, which each have an intron in the conserved region, show strong identity at the amino acid level with the corresponding genes of mammals and chickens. Similar structure and identity of the Sox9 and Sox17 genes among mammals, chickens and fish suggest that these genes have evolutionarily conserved roles, potentially including sex determination and differentiation. PMID- 11929628 TI - Comparative analysis on the structural features of the 5' flanking region of kappa-casein genes from six different species. AB - Kappa-casein plays an essential role in the formation, stabilisation and aggregation of milk micelles. Control of kappa-casein expression reflects this essential role, although an understanding of the mechanisms involved lags behind that of the other milk protein genes. We determined the 5'-flanking sequences for the murine, rabbit and human kappa-casein genes and compared them to the published ruminant sequences. The most conserved region was not the proximal promoter region but an approximately 400 bp long region centred 800 bp upstream of the TATA box. This region contained two highly conserved MGF/STAT5 sites with common spacing relative to each other. In this region, six conserved short stretches of similarity were also found which did not correspond to known transcription factor consensus sites. On the contrary to ruminant and human 5' regulatory sequences, the rabbit and murine 5'-flanking regions did not harbour any kind of repetitive elements. We generated a phylogenetic tree of the six species based on multiple alignment of the kappa-casein sequences. This study identified conserved candidate transcriptional regulatory elements within the kappa-casein gene promoter. PMID- 11929630 TI - Comparison analysis of distributions of tyrosine hydroxylase, calmodulin and calcium/calmodulin-dependent protein kinase II in a triple stained slice of rat brain. AB - The immunohistochemical distributions of tyrosine hydroxylase (TH), calmodulin (CaM) and calcium/CaM-dependent protein kinase II (CaMKII) in the rat forebrain were compared quantitatively to confirm our previous finding that TH activity and dopamine synthesis in the brain are regulated by a calcium/CaM-dependent system. The same slice was triply stained and the above substances were detected immunohistochemically. Their distributions in the slice were measured using a brain mapping analyzer which is a microphotometry system for the analysis of the distribution of neurochemicals in a large tissue slice. Each coronal section was divided into approximately 250000 to 310000 microareas at 20-microm intervals, and the immunohistochemical fluorescence intensities of the three substances in these microareas were analyzed independently. Quantitative images of the distributions were reconstructed from the data, and the distribution of each substance was investigated. Immunoreactive staining of TH, CaM and CaMKII was observed in almost all areas of the brain, but the intensities varied. Markedly intense TH-, CaM- and CaMKII-like immunoreactivities were distributed in the anterior dorsolateral and posterior areas of the neostriatum, nucleus accumbens and olfactory tubercle. In the previous study, the amount of dopamine was increased by the intracerebroventricular administration of calcium chloride in the neostriatum and nucleus accumbens. Combining these results with those previously reported, it is suggested that TH activity and dopamine synthesis in these regions are regulated by calcium ions via CaM and CaMKII. This method is a powerful technique for quantitative and comparative analysis of the distributions of various neurochemicals in the same slice, and we believe that it will facilitate investigation of the functions of the central nervous system and disorders thereof in various diseases. PMID- 11929631 TI - Styrene 7,8-oxide induces caspase activation and regular DNA fragmentation in neuronal cells. AB - Neurobehavioral changes have been described in workers occupationally exposed to styrene vapors. Alterations of neurotransmitters and loss of neurons have been observed in brains of styrene-exposed rats. However, the mechanisms of neuronal damage are not yet clearly understood. We have characterized the cellular alterations induced by the main reactive intermediate of styrene metabolism, styrene 7,8-oxide (SO) in the human neuroblastoma SK-N-MC cell line and primary culture of rat cerebellar granule cells (CGC). SK-N-MC cells exposed to SO (0.3-1 mM) displayed apoptotic morphology, together with chromatin condensation and DNA cleavage into high molecular weight fragments of regular size. These features were accompanied by the activation of class II caspases, as detected with the DEVD assay, by following the cleavage of the caspase-substrate poly (ADP-ribose) polymerase (PARP) and by detection of the active fragment of caspase-3. Pre incubation of the cells with the caspase inhibitor z-VAD-fmk reduced the cellular damage induced by SO, suggesting that caspases play an important role in SO toxicity. Increased proteolysis by class II caspases was detected also in primary culture of CGC exposed to SO. In addition, the presence of the 150-kDa cleavage product of alpha-fodrin suggests a possible activation of calpains in SK-N-MC cells. Moreover, SO did not affect the level of expression of the p53 protein, even though it is known to cause DNA damage. The identified intracellular pathways affected by SO exposure provides end-points that can be used in future studies for the evaluation of the neurotoxic effect of styrene in vivo. PMID- 11929633 TI - Regional alterations of the NO/NOS system in the aging brain: a biochemical, histochemical and immunochemical study in the rat. AB - We have used several approaches (immunohistochemistry and enzyme histochemistry, Western blotting, biochemical assay of Ca(2+)-dependent catalytic activity) in order to detect differences in neuronal nitric oxide synthase (nNOS) expression and activity in various brain regions of young-adult (4-month-old) and aged (28 month-old) rats. In most of the brain regions examined (striatum, neocortex, olfactory cortex and hippocampus) some significant decrease in the density per unit area of nNOS neurons, detected either through immunohistochemistry or enzyme histochemistry, was observed in aged rats. However, only in the striatum and olfactory cortex this was accompanied by a significant decrease of the catalytic activity of the constitutive, Ca(2+)-dependent NOS form. In these two regions, the relative level of expression of nNOS protein was also significantly decreased, as assessed by Western blotting of proteic extracts from young-adult and aged rats. Other observed differences were a paler stain of neurons in some brain areas of the aged rats and differences of cellular compartmentalization of the protein in the same rats, as assessed through confocal microscopy. The present observations demonstrate that the expression and activity of nNOS show regionally-specific alterations in the brain of aged healthy rats, with a trend towards decrease, rather than toward increase as suggested by some previous reports. Therefore, hypotheses implicating nitric oxide increase in brain aging should be reconsidered on the basis of a clear-cut distinction between the physiological and the pathological aspects of the aging process. PMID- 11929632 TI - Nuclear factor kappaB activation is mediated by NMDA and non-NMDA receptor and L type voltage-gated Ca(2+) channel following severe global ischemia in rat hippocampus. AB - Recent studies suggest that nuclear factor NF-kappaB may be involved in excitotoxin-induced cell apopotosis. To analyze the variation of NF-kappaB, levels of NF-kappaB were measured after the rats were subjected to 30 min of four vessel occlusion and sacrificed in selected reperfusion time points. Induction of NF-kappaB consisting mainly of p65 and p50 subunits was detected by Western blot with anti p65, p50 antibodies, respectively. DNA binding activity of NF-kappaB was performed by electrophoretic mobility-shift analysis. Our studies indicate that ischemia-induced NF-kappaB nuclear translocation is time-dependent. Inductions or binding activity of NF-kappaB in nucleus increased about 10-fold from 6 to 12 h as compared with that of the control group, then gradually declined in the following 24, 72 h. To further analyze the regulation by ionotropic glutamate receptor and L-type voltage-gated Ca(2+) channel (L-VGCC) in vivo, N-methyl-D-aspartate (NMDA) receptor antagonist ketamine, alpha-amino-3 hydroxy-5-methyl-4-isoxazolepropionate/kainate receptor antagonist 6,7 dinitroquinoxaline-2,3 (1H,4H)-dione and L-VGCC antagonist nifedipine were given 20 min prior to 30 min of ischemia. The NF-kappaB nuclear translocation was completely blocked by these three antagonists in a dose-dependent manner after ischemia/reperfusion 6 h. Increased phosphorylation of the NF-kappaB regulatory unit IkappaB-alpha was detected by Western blot. Decrement of IkappaB-alpha was found after 3 h reperfusion in the cytoplasm following global ischemia, which was also blocked by such three antagonists. These results illustrate that glutamate gated ionotropic NMDA or non-NMDA receptors and voltage-gated Ca(2+) channels are important routes to mediate NF-kappaB activation during brain ischemic injury. Active NF-kappaB may attend the excitotoxin-induced cell death in turn. Our studies also suggest that IkappaB-alpha is an important regulatory unit that controls the activation of NF-kappaB after its phosphorylation and degradation and resynthesis in rat hippocampus following global ischemia. PMID- 11929634 TI - Stromelysin-1 and gelatinase A are upregulated before TNF-alpha in LPS-stimulated neuroinflammation. AB - Neuroinflammation induces a complex molecular cascade that leads to the proteolysis of cells. Matrix metalloproteinases (MMPs) attack all components of the extracellular matrix in a number of neuroinflammatory diseases and cause a delayed opening of the blood-brain barrier (BBB). Earlier, we showed that lipopolysaccharide (LPS) disrupted the BBB through the action of gelatinase B (MMP-9). In a study of cerebral ischemia, gelatinase A (MMP-2) was seen in astrocytic end-feet and stromelysin-1 (MMP-3) in microglia. Since other MMPs may be important in LPS-induced injury, we studied the gene transcription and cellular localization of several MMPs and an inflammatory mediator, tumor necrosis factor (TNF-alpha), using competitive polymerase chain reaction (PCR) and immunohistochemical methods. Significantly elevated levels of MMP-2 and -3 mRNA were observed in LPS-injected brains by 2 h after injection as compared to non-injected brain tissue (P<0.05). By 8 h post-LPS injection, gene expression of MMP-2 and -3 had declined in both saline- and LPS-injected tissue, while TNF alpha mRNA levels rose significantly. Immunohistochemistry of control brains confirmed the earlier observation of MMP-2 immunoreactivity in processes abutting cerebral blood vessels, which increased after LPS injection. The expression of MMP-9 and MMP-3 was localized mainly to the cerebrovasculature in LPS-stimulated brain tissue, predominantly in the perivascular cells of the basal lamina near the site of injection. Both of these proteinases were present at the site of LPS injection at 8 h, but MMP-2 was absent. Our results show that MMP genes are up regulated prior to the induction of cytokines such as TNF-alpha, and that MMP proteins are prominent around blood vessels in LPS-induced neuroinflammation. PMID- 11929635 TI - Phosphorylation of P42/P44 MAP kinase and DNA fragmentation in the rat perforant pathway stimulation model of limbic epilepsy. AB - The intracellular signaling pathways associated with neuronal injury after perforant pathway stimulation of the rodent hippocampus have not been examined. To determine whether activation of the p42/p44 (Erk1/2) MAP kinase (MAPK) phosphorylation cascade is linked to neuronal injury after perforant pathway stimulation (PPS), we stained for phosphorylated Erk1/2 (P-Erk1/2) and for DNA fragmentation, a marker of cell death after PPS. Eighteen Sprague-Dawley rats underwent PPS for 6 (n=6), 12 (n=6), or 24 (n=6) h and were sacrificed either immediately (n=9) or 48 h (n=9) after stimulation. Sham-operated non-stimulated control animals (n=2) and control animals receiving low frequency stimulation only (n=4) were also examined. Brain sections were stained for DNA fragmentation and P-Erk1/2. DNA fragmentation was evident only in granule cells and CA3 pyramidal cells of the stimulated side 48 h after 24 h of PPS. PPS resulted in robust phosphorylation of Erk1/2 that displayed a stereotyped timecourse, appearing first in hilar neurons on the ipsilateral side and later in hilar neurons, granule cells, hippocampal pyramidal and non-neuronal cell populations on both the stimulated and contralateral sides. Both Erk1/2 phosphorylation and DNA fragmentation show definite and reproducible staining patterns after PPS that vary based on duration of stimulation. Populations displaying Erk1/2 activation appeared to differ from those showing DNA fragmentation and neuronal injury. PMID- 11929636 TI - Pro-inflammatory effects of aluminum in human glioblastoma cells. AB - Inflammatory events have been associated with senile plaques, one of the pathological hallmarks of Alzheimer's disease (AD). It is believed that aggregated beta-amyloid (betaA) proteins, which form the core of these plaques, may be responsible for triggering the inflammatory reaction. In the present study, the ability of aluminum (Al) to initiate similar inflammatory events was investigated in a human glioblastoma cell line. A 6-day exposure to either lipopolysaccharide (LPS) or aluminum sulfate caused a significant increase in the rate of proliferation of the glioblastoma cells. Both treatments also caused activation of the immune-responsive transcription factor NF-kappaB although there were time-related differences. The levels of secreted cytokines, interleukin-6 (IL-6) and tumor necrosis factor alpha (TNF-alpha) were both increased by the LPS treatment although exposure to Al decreased the secretion of the former while elevating the levels of the latter. These events may be due to the activation of glial cells and subsequent stress response to either Al complexes or LPS. Although exposure to either stress factor caused a stimulation of inflammatory markers, there were time-dependent differences in the response. This may reflect the ability of the cells to discern different stress factors and thus orchestrate an innate immune response profile distinct to each immunogen. PMID- 11929637 TI - AMPA glutamatergic receptor-immunoreactive subunits are expressed in lumbosacral neurons of the spinal cord and neurons of the dorsal root and pelvic ganglia controlling pelvic functions in the rat. AB - Sacral preganglionic neurons innervate the pelvic organs via a relay in the major pelvic ganglion. Pudendal motoneurons innervate striated muscles and sphincters of the lower urinary, genital and digestive tracts. The activity of these spinal neurons is regulated by sensory afferents of visceral and somatic origins. Glutamate is released by sensory afferents in the spinal cord, and interacts with a variety of receptor subtypes. The aim of the present study was to investigated the presence of AMPA glutamate receptor subunits (GluR1-GluR4) in the neural network controlling the lower urogenital and digestive tracts of male rats. We performed double-immunohistochemistry directed against a neuronal tracer, the cholera toxin beta subunit (Ctbeta) and each of the four receptor subunits. GluR1, GluR2 and GluR3 subunits were present in many sacral preganglionic neurons retrogradely labelled with Ctbeta applied to the pelvic nerve, and in some dorsolateral and dorsomedian motoneurons retrogradely labelled with Ctbeta injected in ischiocavernosus and bulbospongiosus muscles. The four subunits were detected in postganglionic neurons of the major pelvic ganglion retrogradely labelled with Ctbeta injected in the corpus cavernosum, and in some somata of sensory afferents of the L6 dorsal root ganglion labelled with Ctbeta applied to the dorsal penile nerve or injected in corpus cavernosum. The results provide a detailed knowledge of the neural targets expressing the various AMPA receptor subunits and suggest that part of the neural network that controls pelvic organs, including sensory afferents and postganglionic neurons, is sensitive to glutamate through the whole family of AMPA subunits. PMID- 11929638 TI - Inhibition of neuronal nitric oxide synthase increases adrenalectomy-induced granule cell death in the rat dentate gyrus. AB - Recent studies have shown that the expression of neuronal nitric oxide synthase (NOS) mRNA is increased after adrenalectomy (ADX). However, the role of increased NO production after ADX in the dentate gyrus is unknown. In this study, the relationship between NO inhibition and apoptosis in the dentate gyrus after ADX was examined. 7-Nitroindazole (7-NI; 30 mg/kg, i.p.), a selective inhibitor of neuronal NOS, was injected 1 day before ADX and subsequently once every 24 h. Then 4 days after ADX, dentate granule cell death was evaluated using silver impregnation and Nissl staining methods. Inhibition of neuronal NOS by 7-NI increased the number of dying granule cells approximately 4-fold in the dentate gyrus of the ADX rats, compared to vehicle-injected ADX controls. These results suggest that increased NO production after ADX may play an endogenous neuroprotective role in the dentate gyrus. PMID- 11929639 TI - Reduced nicotinamide nucleotides prevent nitration of tyrosine hydroxylase by peroxynitrite. AB - Tyrosine hydroxylase (TH) is the initial and rate-limiting enzyme in the biosynthesis of the neurotransmitter dopamine (DA). TH activity is inhibited by peroxynitrite (ONOO(-)) by a mechanism that involves nitration of tyrosine residues and oxidation of cysteine residues in the enzyme. Reduced forms of the nicotinamide adenine dinucleotide cofactors, NADH and NADPH, protect TH from inhibition by ONOO(-) and prevent nitration of tyrosine residues. NAD, the oxidized form of the cofactors, neither protects TH from ONOO(-)-induced inhibition nor prevents the nitration of tyrosine residues in the enzyme. These results suggest that the redox status of the nicotinamide nucleotide cofactors could influence the ability of ONOO(-) to modify proteins that are important to the function of DA neurons. PMID- 11929640 TI - Oral contraceptives and venous thromboembolism: a five-year national case-control study. AB - The objective of this study was to assess the influence of oral contraceptives (OCs) on the risk of venous thromboembolism (VTE) in young women. A 5-year case control study including all Danish hospitals was conducted. All women 15-44 years old, suffering a first ever deep venous thrombosis or a first pulmonary embolism (PE) during the period January 1, 1994, to December 30, 1998, were included. Controls were selected annually, 600 per year in 1994-1995 and 1200 per year 1996 1998. Response rates for cases and controls were 87.2% and 89.7%, respectively. After exclusion of nonvalid diagnoses, pregnant women, and women with previous thrombotic disease, 987 cases and 4054 controls were available for analysis. A multivariate, matched analysis was performed. Controls were matched to cases within 1-year age bands. Adjustment was made for confounding influence (if any) from the following variables: age, year, body mass index, length of OC use, family history of VTE, cerebral thrombosis or myocardial infarction, coagulopathies, diabetes, years of schooling, and previous birth. The risk of VTE among current users of OCs was primarily influenced by duration of use, with significantly decreasing odds ratios (OR) over time: <1 year, 7.0 (5.1-9.6); 1-5 years, 3.6 (2.7-4.8); and >5 years, 3.1 (2.5-3.8), all compared with nonusers of OCs. After adjustment for confounders, current use of OCs with second- (levonorgestrel or norgestimate) and third- (desogestrel or gestodene) generation progestins when compared with nonuse resulted in ORs for VTE of 2.9 (2.2-3.8) and 4.0 (3.2-4.9), respectively. After adjusting for progestin types and length of use, the risk decreased significantly with decreasing estrogen dose. With 30-40 microg as reference, 20 and 50 microg products implied ORs of 0.6 (0.4-0.9) and 1.6 (0.9-2.8), respectively (p(trend) = 0.02). After correction for duration of use and differences in estrogen dose, the third/second-generation risk ratio was 1.3 (1.0-1.8; p <0.05). In conclusion, use of OCs was associated significantly to the risk of VTE. The risk among current users was reduced by more than 50% during the first years of use. The risk increased more than 100% with increasing estrogen dose, and the difference in risk between users of third- and second generation OCs, after correction for length of use and estrogen dose, was 33%. PMID- 11929641 TI - Contraceptives and cerebral thrombosis: a five-year national case-control study. AB - The object of this study was to assess the influence of oral contraceptives (OCs) on the risk of cerebral thromboembolic attacks (CTA) including thrombotic stroke and transitory cerebral ischemic attacks. A 5-year case-control study including all Danish hospitals was conducted. All women 15-44 years old suffering a first ever CTA during the period January 1, 1994 to December 31, 1998, were included. Controls were selected annually, 600 per year in 1994-1995, 1200 per year 1996 1998. Response rates for cases and controls were 88% and 90%, respectively. After exclusion of nonvalid diagnoses, pregnant women, and women with previous thrombotic diseases, 626 cases and 4054 controls were available for analysis. A multivariate matched analysis was performed. Controls were matched to cases within 1-year age bands. Adjustments were made for the following potential confounders: year, length of OC use, smoking, hypertension, migraine, family CTA, and years of schooling. There were 212 and 1208 current users of OCs among cases and controls, respectively. The risk of CTA among current users of OCs decreased significantly with decreasing estrogen dose (nonusers reference): OCs with 50 microg, 30-40 microg, 20 microg ethinyl estradiol (EE) and progestin-only pills implied adjusted odds ratios (ORs) (95% CI) of 4.5 (2.6-7.7), 1.6 (1.3-2.0), 1.7 (1.0-3.1), and 1.0 (0.3-3.0), respectively. Current users of OCs with second- (levonorgestrel or norgestimate) and third- (desogestrel or gestodene) generation progestins combined with 20-30 microg EE had ORs of CTA of 2.2 (1.6-3.0) and 1.4 (1.0-1.9), respectively. After correction for differences in estrogen dose, the third- to second-generation risk ratio was 0.6 (0.4-0.9; p = 0.01). In conclusion, high dose OCs and OCs with second-generation progestins were associated with the risk of CTA. The risk increased 2.5 times with estrogen dose increasing from 20 to 50 microg EE, and users of low-dose OCs with second generation progestins had a 61% higher risk-association of CTA than users of OCs with third-generation progestins. PMID- 11929642 TI - Case-control studies on venous thromboembolism: bias due to design? A methodological study on venous thromboembolism and steroid hormone use. AB - This study addressed methodological issues of epidemiologic studies on venous thromboembolism (VTE) to show how design decisions can affect the results. It examines the influence of a restricting to hospitalized and to "idiopathic" cases of VTE in case-control studies on VTE risk and oral contraceptive (OC) use and thereby the bias potential. The diagnostic processes and OC exposures of 1068 suspected cases of VTE were followed in 21 German centers from 1994 to 1999 and evaluated compared to population and hospital controls. Of 606 confirmed cases, 385 (65.5%) were hospitalized, 168 (27.7%) were "idiopathic." Comparing users versus nonusers of OCs, the odds ratio (OR) for VTE risk for all cases and controls was 3.38 and 5.44 for "idiopathic" VTE. For hospital cases and controls, the OR was 3.72 and 9.1 for "idiopathic" VTE. The risk ratio for third- vs. second-generation OCs was increased in the hospital base but not in the population base. It was concluded that restriction to hospitalized events and exclusions of certain cases overestimate the VTE risk of OCs. An evidence-based consensus on methodological standards and definitions in case-control studies on VTE and steroid hormone use is required. PMID- 11929643 TI - An open label, comparative study of the effects of a dose-reduced oral contraceptive containing 20 microg ethinyl estradiol and 100 microg levonorgestrel on hemostatic, lipids, and carbohydrate metabolism variables. AB - In this open label, randomized study we compared the influence of a dose-reduced oral contraceptive containing 20 microg ethinyl estradiol (EE) and 100 microg levonorgestrel (20 EE) with a reference preparation containing 30 microg EE and 150 microg levonorgestrel (30 EE) on hemostatic, lipids, and carbohydrate metabolism variables. Data from 48 volunteers were obtained. The direction of the change (increase or decrease) in most of the hemostatic variables were similar in both treatment groups. In particular, prothrombin fragment 1 + 2 increased during treatment, reaching a median percent change of 40% in the 20 EE group and of 17% in the 30 EE group after one year. D-Dimer fibrin split products remained virtually unchanged, with no change at Cycle 13. The median HDL2 cholesterol levels decreased by 26% in the 20 EE group and by 39.8% in 30 EE group (p = 0.0045 for group difference) after one year. The median one year change for LDL cholesterol was 3.23% in the 20 EE group, compared to 25% in the 30 EE group, for VLDL 11.1% compared to 38.8%, respectively, and for total triglycerides 10.0% compared to 37.5%, respectively. The median absolute change for the area under the curve (AUC)(0-3h) for glucose at treatment Cycle 13 was 41.25 mmol/L x min in the 20 EE group and 73.50 mmol/L x min in the 30 EE group. The AUC(0-3h) insulin at treatment Cycle 13 decreased in the 20 EE group by 1635.0 pmolL x min and increased in the 30 EE group by 11797.5 pmolL x min (p = 0.0491 for group difference). Both study treatments were safe and well tolerated by the volunteers. In conclusion, the balanced one-third dose reduction in this new oral contraceptive evoked similar effects on the hemostatic variables, but favorable results for the lipid and carbohydrate profiles. PMID- 11929644 TI - Effect of dienogest-containing oral contraceptives on lipid metabolism. AB - In a double-blind, controlled, randomized, four-arm, bicentric clinical study, the effect of four oral contraceptives (OCs) on lipid metabolism was investigated. Four groups composed of 25 volunteers each (mean age 26.1 +/- 4.5 years; body mass index 21.9 +/- 2.8 kg/m(2)) were treated for six cycles with monophasic combinations containing 21 tablets with either 30 microg ethinyl estradiol (EE) + 2 mg dienogest (DNG) (30 EE/DNG), 20 microg EE + 2 mg DNG (20 EE/DNG), 10 microg EE + 2 mg estradiol valerate (EV) + 2 mg DNG (EE/EV/DNG), or 20 microg EE + 100 microg levonorgestrel (LNG; EE/LNG). The study was completed by 91 women. Blood samples were taken by venipuncture after at least 12 h fasting on Days 21-26 of the control cycle and Days 18-21 of the first, third, and sixth treatment cycle. There were clear differences between the effects of EE/LNG and the formulations containing estrogens and DNG. Although EE/LNG did not change the triglycerides levels, a significant increase was observed during treatment with the DNG-containing preparations. Although EE/LNG significantly reduced HDL-CH and HDL(2)-CH, there was a nonsignificant increase with the DNG-containing OCs. No change was observed in the levels of HDL(3)-CH. A significant rise in apolipoprotein A1 occurred during intake with the three DNG-containing formulations, but not with EE/LNG. In contrast to the women treated with combinations of estrogens and DNG, apolipoprotein B rose significantly in the women in the EE/LNG group. Lipoprotein (a) was significantly reduced by 30 EE/DNG and EE/LNG and remained unaltered with 20 EE/DNG and EE/EV/DNG. Altogether, the changes in lipid metabolism caused by the DNG-containing formulations appeared to be more favorable than those observed with EE/LNG. In OCs with DNG, the EE dose does not seem to play a major role with respect to the effect on lipids. PMID- 11929645 TI - Long-term effects of combined oral contraceptives on markers of endothelial function and lipids in healthy premenopausal women. AB - The aim of this prospective cross-over study was to investigate the effect of two low-dosed oral contraceptives on markers of endothelial function and plasma lipids. Twelve healthy, nonsmoking women (mean age: 21.7 years) were recruited from the family planning clinic of the university hospital Zurich. For 6 months the participants received a treatment with two contraceptive pills containing 30 microg ethinyl estradiol/150 microg levonorgestrel (three cycles) and 30 microg ethinyl estradiol/75 microg gestodene (three cycles). Plasma levels of endothelin 1, nitric oxide, cholesterol, and HDL were measurement before and during treatment with both oral contraceptive treatments. No significant changes in the plasma levels of nitric oxide and endothelin-1, both important regulators of the vascular tone, were observed during oral contraceptive use. A significant negative correlation was found between nitric oxide and endothelin-1 and nitric oxide and cholesterol. There was a positive correlation between endothelin-1 and cholesterol. In conclusion, the investigated contraceptive pills did not cause major changes in circulating nitric oxide and endothelin-1 plasma levels. PMID- 11929646 TI - Is postpartum contraceptive advice given antenatally of value? AB - In response to the concept that a good postpartum program should begin prenatally, this study was designed to determine whether the provision of expert contraceptive counseling during the antenatal period would have an impact on contraceptive uptake, patterns of contraceptive usage, and pregnancy rates during the first year after childbirth. Over 500 women attending antenatal clinics in each of three centers (Edinburgh, Scotland; Shanghai, People's Republic of China; Cape Town, South Africa) were randomized to receive expert contraceptive advice (participants, n = 771) or the standard advice routinely given in that setting (controls, n = 866). Follow-up was by postal or interviewer-administered questionnaires at 16 and 52 weeks after childbirth. There were no significant differences in the prevalence of contraceptive use at one year (over 79% in all centers) between participants and controls. In Edinburgh, participants were more likely to undergo sterilization (p < 0.01) than controls, otherwise there were no differences among Edinburgh, Shanghai, or Cape Town in either the methods of contraception chosen or in the methods used over time. Contraceptive counseling delivered antenatally appeared to have no impact on the pregnancy rate during the first year after childbirth. In Shanghai, over 11% of women in both groups underwent termination of pregnancy in the year of follow-up. In conclusion, although women in all centers said they found the opportunity to discuss contraception antenatally was useful, it had very little effect on contraceptive use or on subsequent pregnancy rates. PMID- 11929647 TI - Influence of interstitial fibrosis on spermatogenesis after vasectomy and vasovasostomy. AB - This study focused on the testicular interstitial fibrosis after vasectomy and the intraseminiferous status [i.e., Johnsen's score, proliferative cell nuclear antigen (PCNA) expression] following vasectomy reversal. Testicular biopsy specimens from 21 consecutive men were obtained at vasovasostomy. Percent of interstitial fibrosis was determined quantitatively by NIH-image after Masson Trichrome staining. PCNA-labeling index (LI) was calculated on each testis. The associations between the obstructive interval and each parameter were examined. These parameters were also analyzed on whether patency or pregnancy was achieved or not. Significant decrease in PCNA-LI and increase in interstitial fibrosis were observed along with the obstructive interval (p <0.0001, p = 0.0005, respectively). Interstitial fibrosis of the patients without patency/fertility was significantly greater than that of the patients with patency/fertility (47.5%/39.0% versus 33.4%/32.3%, p = 0.02/0.04, respectively). PCNA-LI and Johnsen's score did not predict the treatment outcome. Interstistial fibrosis, but not the extent of germ cell differentiation or DNA synthesis, reflects the treatment outcome after vasectomy reversal. Interstistial fibrosis contributes to the irreversible damage of vasectomized testes. PMID- 11929648 TI - Expression of P16(INK4a) in testis of rhesus monkey during heat stress and testosterone undecanoate induced azoospermia or oligozoospermia. AB - Previous studies on azoospermia or oligozoospermia induced by heat stress or high doses of testosterone mainly focused on germ cell apoptosis; no data regarding their possible effect on spermatogonia mitosis are available. We have established unilateral cryptorchid and testosterone undecanoate (TU)-treated monkey models and examined expression of P16(INK4a) in the testis to look at its possible role in azoospermia or oligozoospermia induced by the heat stress or the TU treatment. The results showed that both heat stress and TU were capable of inducing expression of P16(INK4a) mainly in spermatogonia and other types of germ cells as well as Sertoli cells at the later stage of germ cell apoptosis, namely on Day 10 after operation or on Day 60 after TU injection. It is, therefore, suggested for the first time that P16(INK4a) protein may inhibit the spermatogonia mitosis in the testis at the later stage of the germ cell apoptosis, resulting in arrest of spermatogenesis. PMID- 11929649 TI - Urinary protein excretion and expectant management of early onset, severe pre eclampsia. AB - OBJECTIVE: To evaluate the importance of proteinuria in the expectant management of early onset, severe pre-eclampsia. METHODS: In this prospective series of 340 women, 24-h urine collections were performed and monitored twice weekly in a high care ward. RESULTS: Seventy-four women with at least two 24-h urine collections were grouped into women with a proteinuria increase of > or =2 g (n=29) and with women whose proteinuria decreased, or increased by <2 g (n=45). Major maternal complications, prolongation of gestation, and perinatal outcomes were comparable. Fifty-six (75%) women experienced an increase in proteinuria. When patients with heavy proteinuria (n=83) were compared to those with moderate proteinuria (n=257), maternal and perinatal outcomes were comparable. More days were gained before delivery in the heavy proteinuria group than in the moderate (12 vs. 9; P<0.001). CONCLUSION: Most patients experienced increased proteinuria. Neither the rate of increase nor the amount of proteinuria affected maternal and perinatal outcomes. PMID- 11929650 TI - Risk factors for arrest of descent during the second stage of labor. AB - OBJECTIVE: To define obstetrical risk factors for arrest of descent during the second stage of labor and to determine perinatal outcome. STUDY DESIGN: All singleton, vertex, term deliveries with an unscarred uterus, between the years 1988 and 1999 were included. Univariable and multivariable analysis were performed to investigate independent risk factors associated with arrest of descent during the second stage of labor and the perinatal outcome. RESULTS: The study included 93266 deliveries, of these 1545 (1.7%) were complicated with arrest of descent during the second stage of labor. Using a multivariable analysis, the following obstetric risk factors were found to be significantly associated with arrest of descent: nulliparity (OR=7.8, 95% CI=6.9-8.7; P<0.001), birth weight >4 kg (OR=2.3, 95% CI=1.9-2.8; P<0.001), epidural analgesia (OR=1.8, 95% CI=1.6-2.0; P<0.001), hydramnios (OR=1.6, 95% CI=1.3-2.0; P<0.001), hypertensive disorders (OR=1.5, 95% CI=1.3-1.8; P<0.001), gestational diabetes A1 and A2 (OR=1.5, 95% CI=1.2-1.8; P<0.001), male gender (OR=1.4, 95% CI=1.2-1.5; P<0.001), premature rupture of membranes (PROM, OR=1.3, 95% CI=1.04-1.6; P=0.021), and induction of labor (OR=1.2, 95% CI=1.02-1.4; P=0.030). Deliveries complicated by arrest of descent resulted in cesarean section in 20.6%, vacuum extraction in 74.0%, and forceps delivery in 5.4%. Newborns delivered after arrest of descent during the second stage of labor had significantly higher rates of low Apgar scores (<7) at 1 and 5 min, as compared to the controls (12.7 vs. 2.1%, P<0.001; and 0.9 vs. 0.2%, P<0.001, respectively). Nevertheless, no significant differences were noted between the groups regarding perinatal mortality (0.38 vs. 0.44%; P=0.759). CONCLUSIONS: Major risk factors for arrest of descent during the second stage of labor were nulliparity, fetal macrosomia, epidural analgesia, hydramnios, hypertensive disorders and gestational diabetes mellitus. These risk factors should be carefully evaluated during pregnancy in order to actively manage high-risk pregnancies. PMID- 11929651 TI - Induction of labor in twin pregnancy after 36 weeks does not increase maternal fetal morbidity. AB - OBJECTIVE: To compare maternal and perinatal outcomes in twin pregnancy patients without any fetal or maternal disease, managed by labor induction or expectant management after 36 weeks gestation. METHODS: We conducted a case-control study of 81 patients: labor induction (n=36) vs. expectant management (n=45). Labor was induced using oxytocin (n=18), vaginal prostaglandins (n=6) or intrauterine balloon catheter (n=12) according to the Bishop score. Maternal and perinatal outcome variables were compared among both groups. RESULTS: The characteristics of the labor induction group and the expectant management group were not statistically different, except for the rate of nulliparae (55.6% vs. 33.3%) and the rate of epidural analgesia (100% vs. 80%). There was no significant difference in labor time (6.5+/-2.8 h vs. 6.0+/-3.6 h), cesarean section rate (8.3% vs. 13.3%) or duration of maternal hospitalization (7.3+/-2.0 days vs. 7.5+/-2.3 days) in the labor induction group and in the expectant management group, respectively. The birth weight was higher in the labor induction group than in the expectant management group (2639+/-352 g vs. 2463+/-298 g, P<0.001). The rate of Apgar score <7 at 5 min was 0% and 3.3%, respectively. Neonatal intensive care unit admission occurred in 30.5% and 26.6% of the groups, respectively. No perinatal death was reported. CONCLUSION: Induction of labor may be proposed to patients with uneventful twin pregnancy after 36 weeks gestation without increasing maternal-fetal morbidity. PMID- 11929652 TI - Adequacy of oophorectomy at the time of gynecologic surgery. AB - OBJECTIVES: To determine the incidence of incomplete ovarian removal during gynecologic surgery and correlate the risk of inadequate removal with the procedure chosen. METHODS: This is a prospective observational study. Ovaries received during a 4-month period in the participating institutions were independently histologically evaluated. Gross inspection of the ovarian capsule, infundibulopelvic ligament, hilum and utero-ovarian ligament was assessed. Grossly close margins were confirmed histopathologically. Any margin with histologically confirmed ovarian tissue at the margin was interpreted as incompletely removed. Details of each surgical procedure were recorded for comparison. RESULTS: Ovaries (n=174) from 94 patients were collected and 155 were evaluable. The overall incidence of incomplete ovarian removal was 6.5%. Of the 125 ovaries removed abdominally, 23 were laparoscopically assisted and 7 were vaginal; inadequate removal was documented in 5%, 9% and 29%, respectively (P=0.04). There was no relationship of inadequate resection by underlying pathologic diagnosis (P=0.25) or by institution (4.6% university hospital vs. 8.8% community hospital; P=0.29). CONCLUSIONS: Incomplete ovarian removal occurs and is related to surgical approach. A larger study is warranted to evaluate the role of pelvic pathology or surgeon experience as a risk for incomplete oophorectomy. PMID- 11929653 TI - Vaginal birth after cesarean section in a rural African setting. PMID- 11929654 TI - Angiotensin-converting enzyme activity in pre-eclampsia. PMID- 11929655 TI - Maternal serum uric acid as predictor of fetal acidemia. PMID- 11929656 TI - Intrauterine adhesions in an African population. PMID- 11929657 TI - Screening for hypothyroidism at a menopause clinic. PMID- 11929658 TI - Methemoglobinemia following chromopertubation in treated pelvic tuberculosis. AB - Methylene blue administered through cervix is used to check tubal patency at laparoscopy. Although methylene blue is used to treat methemoglobinemia it can also produce methemoglobinemia in susceptible individuals. A case of methemoglobinemia induced by methylene blue in a patient with treated pelvic tuberculosis is presented. PMID- 11929659 TI - A comparison of the prevalence of domestic violence between patients seeking termination of pregnancy and other general gynecology patients. AB - OBJECTIVES: (1) To determine and compare the prevalence of domestic violence among abortion-seeking patients with other general gynecology patients; (2) to see if a follow-up interview 6 weeks after abortion can improve the abuse disclosure rate; (3) to see if the abused victims accept direct referral to their gynecologists/social workers for help. METHODS: This is a prospective questionnaire survey in a university teaching hospital on patients seeking abortion and an approximately equal number of other general gynecology patients. Participants were interviewed by a designated project nurse in a private setting, using a structured questionnaire (Modified Abuse Assessment Screen Questionnaire) to assess the past and recent history of abuse. The interview was repeated 6 weeks after the abortion for the abortion-seeking group. RESULTS: Five hundred and one participants were interviewed, including 245 seeking abortion (TOP group) and 256 other general gynecology patients (non-TOP group). The lifetime prevalence of abuse in the TOP group (27.3%) was significantly higher than the non-TOP group (8.2%) (P<0.001). Repeating the interview 6 weeks after the abortion did not increase the disclosure rate. Most abused victims were unwilling to disclose their information of abuse to their gynecologists or social workers at the time of interview. CONCLUSIONS: Domestic violence is a significant problem among the gynecology patients, particularly those seeking abortion. A single interview prior to abortion is adequately effective for screening. However, the most effective and acceptable way of helping these victims needs to be explored further. PMID- 11929660 TI - Human rights and HIV-positive women. AB - Recent court decisions, for instance in South Africa and Latin America, have held states bound to respect and serve HIV/AIDS patients' human rights to indicated and available medical care. HIV/AIDS is estimated to affect over 36 million people worldwide, including 16.4 million women of reproductive age. In the last 20 years, nearly 58 million people have been infected. This article reviews national responses to mounting concern with the HIV/AIDS pandemic, particularly in China, India and Africa, medical professional responses, notably by the World Medical Association, and international guidelines on human rights responses. These pay special attention to patients' rights to be treated without discrimination. It addresses national and international approaches to advancing HIV prevention, treatment and research on which UNAIDS and the UN High Commissioner for Human Rights have collaborated. Special issues in clinical care concern abortion services for HIV-positive women, breastfeeding and patients' involvement in research. PMID- 11929661 TI - Stability indicating methods for the determination of diloxanide furoate. AB - Five new selective, precise and accurate methods are described for the determination of diloxanide furoate (DI) in presence of its degradation products. Method A utilizes the first and second derivative spectrophotometry at 270 and 280 nm, respectively. Method B is a RSD(1) spectrophotometric method based on the simultaneous use of the first derivative of ratio spectra and measurement at 270 nm. Method C is a pH-induced difference spectrophotometry using UV measurement at 295 nm. Method D is a densitometric one, after separation on silica gel plate using chloroform: methanol as mobile phase and the spots were scanned at 258 nm. Method E is reversed phase high performance liquid chromatography using methanol: water (80:20% v/v) as mobile phase at a flow rate of 1 ml/min and UV detection at 258 nm. Regression analysis showed good correlation in the concentration ranges 5 30, 5-25, 10-40 microg/ml, 100-500 ng/spot, 2-50 microg/ml with percentage recoveries of 99.92+/-0.56 and 99.79+/-0.47, 99.23+/-0.38, 99.96+/-0.06, 99.03+/ 0.51, 98.81+/-0.68 for methods A, B, C, D and E, respectively. These methods are suitable as stability indicating methods for the determination of DI in presence of its degradation products either in bulk powder or in pharmaceutical formulations. PMID- 11929662 TI - Determination of travoprost and travoprost free acid in human plasma by electrospray HPLC/MS/MS. AB - A quantitative method for the analysis of AL-5848, the (+)-enantiomer of fluprostenol (FP), in human plasma is described. Plasma was spiked with a tetradeuterated analog of travoprost free acid (AL-5848X) as internal standard (IS) and acidified with 0.1 M formic acid. Sample clean up was performed using reversed phase solid-phase extraction. Following elution of the compounds of interest and evaporation to dryness, the residue was reconstituted in methanol:water (1:1) and chromatographed on an octadecylsilica (C18) column with negative ion electrospray ionization tandem mass spectrometry. The [M[bond]H](-) ions at m/z 457 and 461 for the analyte and IS, respectively, were subjected to collisional fragmentation with argon to yield the same intense 3 trifluoromethylphenolate (m/z 161) product ion. The validated concentration range was 0.010-3.00 ng/ml based on a 1.0 ml plasma aliquot. Fully adequate accuracy, precision, specificity, recovery and stability for routine use in clinical pharmacokinetic studies were demonstrated. Analysis of a second plasma aliquot following incubation with rabbit esterase allows the isopropyl ester pro-drug, travoprost (AL-6221), to be determined by difference. PMID- 11929663 TI - Rapid microtiter plate assay for determination of inulin in human plasma and dialysate. AB - A rapid, sensitive, and reproducible microtiter plate assay for the determination of inulin in human plasma, dialysate, and phosphate-buffered saline (PBS) was developed. Plasma or PBS samples (100 microl aliquots) were prepared by the addition of indole-3-acetic acid (150 microl) and HCl (3 ml) and then briefly vortex-mixed. Samples were then incubated in a 60 degrees C water bath for 20 min, cooled in a room temperature water bath for 40 min, then diluted with deionized, distilled water (DDW; 3 ml) and again vortex-mixed. Finally, an aliquot (200 microl) of each sample was transferred to a 96-well microtiter plate and read spectrophotometrically at 490 nm. Dialysate samples were processed in a similar manner, but required an initial enzymatic step in order to remove dextrose and minimize assay interference. Samples (100 microl aliquots) were prepared by the addition of glucose oxidase/catalase solution (100 microl), briefly vortex mixed, and then incubated in a 37 degrees C water bath for 60 min, samples were then reacted with indole-3-acetic acid as before. Calibration curves were linear over the concentration range of 0.5-4 mg/ml or 0.025-0.4 mg/ml for plasma or PBS and dialysate, respectively; correlation coefficients (r(2)) were >0.99. The intra- and inter-day coefficients of variation in plasma, PBS, and dialysate were <15%. This method is well suited for the rapid analysis of large numbers of samples and is currently being used for in vitro investigations of solute removal by hemodialysis. PMID- 11929664 TI - Voltammetric studies of the interaction of lumazine with cyclodextrins and DNA. AB - The interaction of lumazine, an antibacterial drug, with alpha-, beta cyclodextrins and DNA in aqueous solution was studied by differential pulse stripping voltammetry and cyclic voltammetry as well as UV-vis spectroscopy. The electrochemical and absorption spectral data indicated a 1:1 complex formation of lumazine with alpha-, beta-cyclodextrins and DNA. The nature of the process, taking place at the hanging mercury drop electrode, was clarified. It was found that the complexation of lumazine molecules enhances the stacking interactions which might facilitate the formation of a perpendicularly stacked layer of lumazine-alpha-cyclodextrin complex on the electrode surface. Based on the variations in the current or absorbance, the formation constants and consequently, the Gibbs energy of these complexes were determined. The small size cavity of alpha-cyclodextrin was found to have a greater affinity for lumazine than the beta-cyclodextrin. Moreover, the interactions of lumazine-alpha cyclodextrin or lumazine-beta-cyclodextrin inclusion complex with DNA have been investigated by means of voltammetry. The results suggest that lumazine displayed high affinity for DNA and the inclusion complex decomposed when it binds to DNA. PMID- 11929665 TI - Densitometric determination of diclofenac, 1-(2,6-dichlorophenyl)indolin-2-one and indolin-2-one in pharmaceutical preparations and model solutions. AB - A chromatographic-densitometric method for identification and quantitation of diclofenac and its impurities, i.e. 1-(2,6-dichlorophenyl)indolin-2-one and indolin-2-one in pharmaceutical preparations and model solutions was developed. The effect of pH, temperature and ultra violet (UV) radiation on diclofenac's concentration was investigated. Chromatographic separation was performed on TLC silica gel coated plates with the mobile phase: cyclohexane-chloroform-methanol (12:6:1, v/v/v). Densitometric detection was carried out in UV at lambda=248 nm. The conditions for good separation and the detection limit were established. The recovery for diclofenac was 99.20%, for 1-(2,6-dichlorophenyl)indolin-2-one- 92.34% and for indolin-2-one--95.85%. The method was used for quality assessment of diclofenac in pharmaceutical preparations. Reliable results comparable to those determined by high performance liquid chromatography (HPLC) were obtained. PMID- 11929666 TI - Development and validation of a photometric titration method for the quantitation of sodium chondroitin sulfate (bovine) in Cosequin DS chewable tablet. AB - A photometric titration method was developed and validated to quantitate sodium chondroitin sulfate in raw materials and Cosequin DS chewable tablet(1). About 0.1% (w/v) cetylpyridinium chloride was used to titrate sodium chondroitin sulfate with photometric indication at wavelength 420 nm. The standard curves for sodium chondroitin sulfate showed linearity (r> or = 0.99) over the selected concentration range from 0.6 to 1.4 mg/ml. The chewable tablet was ground to fine powder and extracted with water and the resulting solutions filtered through a 0.45 microm membrane filter. Recovery between 97 and 103%,. The intra- and inter- day precision as indicated by the relative standard deviation (R.S.D.) were not greater than 0.33 and 0.78%, respectively. The method was found to be specific and with excellent linearity, accuracy and precision and is well suited for the quantitation of sodium chondroitin sulfate in raw material and Cosequin DS chewable tablet. PMID- 11929667 TI - Validation of a near-infrared transmission spectroscopic procedure, part A: validation protocols. AB - It is possible to devise calibration and validation protocols that enable the ICH guidelines to conform to the specialized requirements of the NIR method of analysis. Some of the required characteristics of evaluation specified by the guidelines, such as accuracy and repeatability, can be applied directly, just as with any other analytical method. Other characteristics are adapted through the novel use of specialized statistics, or through the use of creative methods and procedures to match the recommendations of the guidelines to the unique and specialized requirements of the NIR method. PMID- 11929668 TI - Simultaneous analysis of dehydroacetic acid, benzoic acid, sorbic acid and salicylic acid in cosmetic products by solid-phase extraction and high performance liquid chromatography. AB - A high-performance liquid chromatographic (HPLC) method for simultaneous determination of dehydroacetic acid (DHA), benzoic acid (BA), sorbic acid (SOA) and salicylic acid (SA) was developed for application to cosmetic products. Isocratic reversed-phase HPLC was employed for quantitative analysis using tetra n-butylammonium (TBA) hydroxide as an ion-pair reagent. Cosmetic samples were purified by solid-phase extraction using Bond-Elut SI cartridges. Four acidic preservatives were eluted with methanol from cartridges. The HPLC assay was carried out using TSK gel ODS-80TM column (5 microm, 150 x 4.6 mm I.D.). The mobile phase consisted of a mixture of water and methanol (65:35, v/v) containing 2.5 mM TBA hydroxide adjusted with phosphoric acid to pH 7.0. The calibration curves of these preservatives showed good linearity with UV detection (235 nm). The correlation coefficients were better than 0.999 in all cases. The lower limits of detection (defined as a signal-to-noise ratio of about 3) were approximately 2.5 ng for DHA, 4.0 ng for BA, 2.0 ng for SOA and 5.5 ng for SA. The procedure described here is simple, selective and is suitable for quality control of finished cosmetic products. PMID- 11929669 TI - An equivalent length model of microdialysis sampling. AB - One of the critical issues in microdialysis sampling is how to predict the extraction fraction (E(d)), based on transport properties of analytes in both tissues and probes. A one-dimensional (1-D) model has been used widely in previous studies to predict E(d) at the steady state. However, this model is valid only for long probes. To this end, an equivalent length (EL) model was developed for probes with any length used in experiments. The key idea in the model was to replace the probe length (L) in the 1-D model with an equivalent length (L(E)) when calculating transport resistance in surrounding tissues. The length difference, (L(E)-L), was assumed to be proportional to the penetration depth of analytes (Gamma). The proportionality constant (lambda) was determined through minimizing the errors in predicted E(d). We found that, the EL model could accurately predict E(d) when lambda=0.369. The maximum error in EL model predictions was <6%, for model constants varying in the same ranges as those in microdialysis experiments. This error was one order of magnitude smaller than that in 1-D model predictions. PMID- 11929670 TI - Comparison of SPE and fast LC to eliminate mass spectrometric matrix effects from microsomal incubation products. AB - Twenty-seven highly diversified pharmaceutical compounds were used as a test set to evaluate matrix effects from microsomal media on LC/MS analyses. The individual effects of Tris buffer, NADPH and microsomes on ESI response were investigated. Direct flow injection MS/MS analysis, using no sample preparation or HPLC separation, gave an average of 2.2-5-fold matrix suppression in MS response from Tris buffer and NADPH. More polar analytes were affected the greatest. To reduce the loss in response, an automated solid phase extraction (SPE) procedure was developed. A much smaller average matrix effect was observed when samples were prepared using a Waters Oasis HLB 96-well SPE. As little as 1 ml of methanol (MeOH) was sufficient to elute most compounds with more than 80% recovery. Comparable results were obtained by directly injecting a protein precipitated incubation onto a fast gradient LC separation prior to MS/MS detection. No advantage was seen by using both SPE and a fast LC separation prior to MS/MS analysis. PMID- 11929671 TI - Chiral high-performance liquid chromatographic analysis of the enantiomers of XK469, a new antitumor agent, in plasma and urine. AB - XK469 (NSC697887), (+/-)-2-[4-(7-Chloro-2-quinoxaliny)oxy]-phenoxy propionic acid, an analog of the herbicide Assure(R), which possesses antitumor activity, especially against murine solid tumors and human xenografts, has recently been found to be the first topoisomerase II beta poison. Both R(+) and S(-) isomers are cytotoxic, although the R-isomer is more potent. A chiral high-performance liquid chromatography (HPLC) assay that utilizes Chirobiotic T column for the measurement of enantiomers of XK469 in plasma has been developed with a limit of quantitation (LOQ) of 0.2 microg/ml using a 0.2 ml plasma sample. Chloroqinoxaline sulfonamide (CQS) was used as the internal standard and the assay has been validated in rat plasma. The within-run coefficient of variations (CVs) were 5.9, 5.0, and 3.1% for the S-isomer and 8.1, 4.2, 6.4% for R(+)-XK469 at 0.2, 1, and 2 microg/ml, respectively. The between-run CVs were 10.5, 5.3, and 1.9% for S(-)- and 10.9, 6.3, and 3.6% for R(+)-XK469. Using this chiral assay, a plasma concentration time data of R(+)-,S(-)-XK469 in a Fischer 344 rat receiving i.v. dosing of S(-)XK469 at 10 mg/kg was monitored. S(-)XK469 was found to be significantly converted to the R-enantiomer in circulation even when the S enantiomer was administered. The predominant inversion from S(-)- to R(+)-XK469 was also observed in the mouse and dog plasma. In the rat, the plasma concentration-time profiles for both isomers follow two compartmental pharmacokinetics with the t(1/2 beta) for the R-enantiomer slightly longer and the clearance of the S-enantiomer higher than the R-enantiomer. PMID- 11929672 TI - A chromatographic method for the quantification of prostaglandin E(1) and prostaglandin A(1) encapsulated in an intravenous lipid formulation. AB - Peripheral vascular disease is a common ailment of the aged and diabetic communities. As the numbers of these individuals increase, the need for therapeutic interventions will continue to grow. One of the possible therapies is the use of prostaglandins (PGE(1), prostacyclin and Iloprost) to decrease the vascular tone and increase vascular blood flow. Due to the hydrophobicity of the prostaglandins and prostaglandin analogues, various vehicles have been utilized to maintain the active pharmaceutical ingredient in a stable solution, e.g. alpha cyclodextrin (Alprostadil, Edex) or emulsified lipid vehicles. In our laboratory, we designed a method for separating and assaying lipid-encapsulated PGE(1). Utilizing organic extraction, automated solid-phase extraction and precipitation techniques, we validated the measurement of the PGE(1) and PGA(1) content of the clinical drug formulation in the microgram per milliliter concentration range with an high performance liquid chromatography (HPLC) assay. PMID- 11929673 TI - Determination of propylthiouracil using 1,3-dibromo-5,5-dimethylhydantoin (DBH): analytical methods of pharmacopeias with DBH in respect to environmental and economical concern: part 9. AB - USP 2000 [The United States Pharmacopeia, Rockville USA, 24th ed., 2000, pp. 1436] and PH. EUR. 1997 [European Pharmacopoeia, third ed., Council of Europe, Strasbourg, 1997, p. 1401] determine propylthiouracil using neutralization titration, whereby 0.1 M silver nitrate and twice boiling is necessary. With the application of 1,3-dibromo-5,5-dimethylhydantoin (DBH), the assay of propylthiouracil can be performed easily, faster and friendlier to environment. A mean deviation of 0.03% and a relative standard deviation of 0.3% are obtained. 5 Bromo-6-propyluracil is formed, when propylthiouracil is determined with DBH. PMID- 11929674 TI - Spectrophotometric and AAS determination of ramipril and enalapril through ternary complex formation. AB - Two sensitive, spectrophotometric and atomic absorption spectrometric procedures are developed for the determination of two antihypertensive agents (enalapril maleate and ramipril). The spectrophotometric procedures for the two cited drugs are based on ternary complex formation. The first ternary complex (copper(II), eosin, and enalapril) was estimated by two methods; the first depends on its extraction with chloroform measuring at 533.4 nm. Beer's law was obeyed in concentration range from 56 to 112 microg ml(-1). The second method for the same complex depends on its direct measurement after addition of methylcellulose as surfactant at the pH value 5 at 558.8 nm. The concentration range is from 19 to 32 microg ml(-1). The second ternary complex (iron(III), thiocyanate, and ramipril) was extracted with methylene chloride, measuring at 436.6 nm, with a concentration range 60-132 microg ml(-1). The direct atomic absorption spectrometric method through the quantitative determination of copper or iron content of the complex was also investigated for the purpose of enhancing the sensitivity of the determination. The spectrophotometric and atomic absorption spectrometric procedures hold their accuracy and precision well when applied to the determination of ramipril and enalapril dosage forms. PMID- 11929675 TI - In vitro stability analyses as a model for metabolism of ferromagnetic particles (Clariscan), a contrast agent for magnetic resonance imaging. AB - Clariscan is a new contrast agent for magnetic resonance imaging. It is an aqueous suspension of ferromagnetic particles injected for blood pool and liver imaging. Previous experiments showed that particles made of 59Fe were taken up by the mononuclear phagocytic system and then solubilised. The present work aims at explaining a possible mechanism for the dissolution of these ferromagnetic particles in the body. The particles were diluted in 10-mM citrate or 10-mM acetate buffers at pH 4.5, 5.0 and 5.5 and incubated at 37 degrees C for up to 22 days, protected from light. The mixtures were analysed at different times during this incubation period using photon correlation spectroscopy, magnetic relaxation, visible spectroscopy and reactivity of the iron with the chelator, bathophenanthroline disulphonic acid. The data obtained with these techniques showed that the particles were almost completely solubilised within 4-7 days when incubated in 10 mM citrate, pH 4.5. Incubation in 10 mM citrate buffer, pH 5.0 revealed a slower solubilisation of the particles, as the changes observed after 72 h of incubation at pH 5.0 were 43-71% of the changes observed at pH 4.5. Incubation in 10 mM citrate, pH 5.5 revealed an even slower solubilisation of the particles, as the changes observed after 72 h of incubation at pH 5.5 were 12-34% of those observed at pH 4.5. Incubation of the particles in 10 mM acetate at pH 4.5, 5.0 and 5.5, as well as incubation of the particles in water pH adjusted to pH 5.1, resulted in only minor or no solubilisation of the particles. The results indicate that the low pH of endosomes and lysosomes, as well as endogenous iron complexing substances, may be important for the solubilisation of these ferromagnetic particles following i.v. injection of Clariscan. PMID- 11929676 TI - Fluorimetric determination of isoxsuprine hydrochloride in pharmaceuticals and biological fluids. AB - Two simple and highly sensitive fluorimetric methods have been developed for the determination of isoxsuprine hydrochloride in bulk, in dosage forms and in biological fluids. The first method involves the direct measurement of the native fluorescence of the drug in the concentration range 0.4-4.0 microg ml(-1), the second method is based on the oxidation of isoxsuprine HCl with cerium(IV) followed by fluorimetric measurement in the concentration range 0.02-0.2 microg ml(-1). The average % found were 99.9 +/- 0.78 and 100.0 +/- 0.62 for the two methods, respectively. The minimum detectability (3 S(B)) were 0.11 and 0.007 microg ml(-1) for the two methods, respectively. The methods results showed insignificant difference with those of the official method. PMID- 11929677 TI - Determination of selenium sulfide using 1,3-dibromo-5,5-dimethylhydantoin (DBH). Analytical methods of pharmacopeias with DBH in respect to environmental and economical concern Part 11. AB - USP 2000 (The United States Pharmacopeia, Rockville, MD, 24th ed., 2000, pp. 1514 1515) and PH. EUR. 1997 (European Pharmacopoeia, 3rd ed., Council of Europe, Strasbourg, 1997, p. 1459) dissolve selenium sulfide by boiling with fuming nitric acid for about 1 h. After cooling to room temperature and dilution with water nitrous acid and nitrogen oxides are removed with urea also by boiling before the iodometric titration is performed. This method can be importantly simplified and improved, when using 1,3-dibromo-5,5-dimethylhydantoin (DBH) in glacial acetic acid in presence of solid potassium bromide. Selenium sulfide is dissolved during 5-10 min at room temperature. The excess of DBH and bromine can be removed with 5-sulfosalicylic acid also without boiling. The point of the indicator change for the iodometric titration is improved in comparison to the method of the pharmacopeias, because the colloidal solution of selenium mostly coagulates. Also the sulfur content of selenium sulfide can be determined with DBH. PMID- 11929678 TI - Comparison of electrochemical detection of acetylcholine-induced nitric oxide release (NO) and contractile force measurement of rabbit isolated carotid artery endothelium. AB - Since the identification of nitric oxide (NO) as an endothelial-derived relaxing factor, it became very important to quantify NO in biological models eventhough it is present in very low concentrations with a very short half-life. The use of electrochemistry as an alternative detection method is quite promising and electrochemical probes are now being developed to detect NO. This paper consists of an amperometric, bi-polymer modified, platinum-iridium microelectrode (Pt 90% Ir 10% alloy, multistranded, total diameter 130 microm) design and its application for NO detection in acetylcholine (Ach) introduced, rabbit isolated carotid artery endothelium model. In a pH range of 3.0-10.0. pH 3.0 was found to be the optimum pH. As the pH values increased up to 10.0, the response current decreased as the oxidation of NO is catalyzed by H(+) in the acidic media. Temperature effect was checked at 25 degrees C (room temperature), 30 and 40 degrees C. An increasing trend was observed in sensor response with the increasing temperature. Most common biological interferences as ascorbic acid, uric acid and glucose were eliminated via bi-polymer coatings of four layers of Nafion and a layer of 50 mM o-phenylenediamine (OPD). When S/N ratio was accepted as 3, limit of detection was calculated as 15 nM. NO release from carotid artery endothelium was also determined by measuring response force in thermostatic isolated organel baths. Obtained force responses (mg) were compared with the electrochemical (nA) sensor responses. PMID- 11929679 TI - Polarographic determination of berberine in the presence of H(2)O(2) in medicinal plants. AB - The mechanism of the parallel catalytic wave of berberine in the presence of H(2)O(2) was studied. The results showed that the reduction process of the C[double bond]N bond of berberine was two successive one-electron transfers, in which an intermediate free radical was involved. When H(2)O(2) was present, it oxidized the free radical to the original C[double bond]N bond, producing the parallel catalytic wave of berberine. In Na(2)B(4)O(7)-Na(2)CO(3) (pH 9.4, 0.08 M)-H(2)O(2)(4 mM) supporting electrolyte, the peak current of the catalytic wave was linear to the berberine concentration in the range of 1.0 x 10(-8)-3.0 x 10( 7) M. The limit of detection was 7.0 x 10(-9) M. The catalytic wave can be applied to direct determination of berberine in medicinal plant Coptis chinensis Franch, after sufficiently diluting the water extraction without preliminary separation. PMID- 11929680 TI - Spectrophotometric, spectrofluorimetric, HPLC and CZE determination of mirtazapine in pharmaceutical tablets. AB - Four analytical methods have been developed for the quality control of tablets containing mirtazapine: spectrophotometry, spectrofluorimetry, high performance liquid chromatography (HPLC) and capillary zone electrophoresis (CZE). All the methods only require a simple extraction procedure of mirtazapine from the tablets before analysis. The concentration of mirtazapine in solutions was determined in the linearity range of 5-25 microg/ml at lambda=315 nm for spectrophotometry and at lambda=220 nm for HPLC and CZE. Spectrofluorimetric determinations were achieved at lambda(excitation)=328 nm and lambda(emission)=415 nm in the linearity range of 2-25 ng/ml. All the methods gave similar results and were validated for selectivity, linearity, precision and sensitivity. Spectrometric methods gave slightly higher RSD values (up to 2.54%). The four methods were directly and easily applied to the pharmaceutical preparation with accuracy, resulting from recovery experiments between 99.72% in HPLC and 101.47% in spectrofluorimetry. PMID- 11929682 TI - Determination of organic acids in leaves and rinds of Garcinia indica (Desr.) by LC. AB - Organic acids in leaves and rinds of Garcinia indica (Kokam) were determined by high-performance liquid chromatography. The major organic acid in leaves and rinds has been found to be (-)-hydroxycitric acid present to the extent of 4.1 4.6 and 10.3-12.7%, respectively, by isocratic elution with 8 mM sulfuric acid as mobile phase with a flow rate of 1.0 ml/min using UV detection at 210 nm. Hydroxycitric acid lactone and citric acid are present in leaves and rinds in minor quantities. This method has been shown to be very reproducible with the coefficient of variation ranging from 2.8 to 4.2%. This is the first report on the composition of organic acids in the leaves and rinds of G. indica by HPLC. PMID- 11929681 TI - Spectrophotometric determination of meclozine HCl and papaverine HCl in their pharmaceutical formulations. AB - A simple, accurate and highly sensitive spectrophotometric method is proposed for the rapid determination of meclozine and papaverine hydrochlorides using chromotrope 2B (C2B) and chromotrope 2R (C2R). The method consists of extracting the formed ion-associates into chloroform in the case of meclozine HCl and into methylene chloride in case of papaverine HCl. The ion-associates exhibit absorption maxima at 536 and 524 nm for C2B and C2R with meclozine HCl and at 540 and 528 nm with papaverine HCl, respectively. Meclozine can be determined up to 4.0 and 2.6 mg ml(-1), using C2B and C2R, respectively, while papaverine can be determined up to 1.68 and 1.37 mg ml(-1), respectively. The effect of acidity, reagent concentration, time, solvent and stoichiometric ratio of the ion associates were studied. The molar absorptivity and Sandell sensitivity of the reaction products were calculated. The method was applied to the determination of the drugs in their pure state or pharmaceutical preparations with mean recovery values of 99.63-100.80 and 99.75-100.08% and coefficient of variation 0.945-2.210 and 1.020-1.268 for meclozine HCl and papaverine HCl, respectively. PMID- 11929683 TI - Separation of N-alkyl phenothiazine sulfones by HPTLC using an optimum mobile phase. AB - In high performance thin layer chromatography some form of optimization is necessary if complete separation of all components is required. The selection of mobile phase composition is one of the most important components of an optimization strategies. The aim of this paper is the separation of the N-alkyl phenothiazine sulfones by high performance thin layer chromatography using an optimum mobile phase system. The optimum composition of mobile phase (toluene ethyl ether-chloroform, 30:50:20, v/v) are given by the maximum of objective function (F(obj)=10.6110). PMID- 11929684 TI - Simultaneous determination of dorzolamide HCL and timolol maleate in eye drops by two different spectroscopic methods. AB - Two-component mixtures of dorzolamide hydrochloride and timolol maleate were assayed by first derivative and ratio derivative spectrophotometric methods. The first method, derivative spectrophotometry, by the zero-crossing measurements, was used due to the drugs closely overlapping absorption spectra. Linear calibration graphs of first derivative values at 250.3 nm for dorzolamide hydrochloride and 315.8 nm for timolol maleate. The second method, is based on ratio first derivative spectrophotometry, the amplitudes in the first derivative of the ratio spectra at 242.9 and at 223.5 nm were selected to determine dorzolamide and timolol maleate in the binary mixture. Calibration graphs were established for 8.0-30.0 microg ml(-1) for dorzolamide hydrochloride and 3.0-24.6 microg ml(-1) for timolol maleate in binary mixture. Good linearity, precision and selectivity were found, and the proposed methods were applied successfully to the pharmaceutical dosage from containing the above-mentioned drug combination without any interference by the excipients. Vierordt's method was also developed for a comparison method. PMID- 11929685 TI - Chlamydia pneumoniae, systemic inflammation and the risk of venous thrombosis. AB - Inflammatory mediators are involved in activation of the coagulation system, and elevated plasma concentrations of IL-6 and IL-8 are associated with an increased risk of venous thrombosis. Using serologic and molecular biologic tests, we investigated in a case-control study on patients with recurrent venous thrombosis the association between Chlamydia (C) pneumoniae and venous thrombosis and we evaluated the relation between C. pneumoniae serology and the cytokines IL-6 and IL-8. The presence of C. pneumoniae antibody titers > or = 1:16 was not associated with an increased risk of venous thrombosis (odds ratio 0.8 95% CI, 0.4-1.7). Circulating C. pneumoniae-DNA was detected in only one patient and two control subjects. IgG antibody titers against C. pneumoniae were not correlated with the concentrations of IL-6 and IL-8. These results indicate that the inflammatory process shown in patients with venous thrombosis is not related to C. pneumoniae. PMID- 11929686 TI - Chronic hepatosplenic abscesses in Brucellosis. Clinico-therapeutic features and molecular diagnostic approach. AB - In order to analyze the clinical and therapeutic features of chronic hepatosplenic abscesses, and to define the diagnostic yield of new molecular techniques, we describe seven cases, four hepatic and three splenic, of this uncommon complication of Brucellosis. Onset of symptoms in all cases was insidious and the diagnostic delay considerable. Abdominal CT scan showed large, poorly defined lesions, with heterogeneous attenuation and thick central calcifications surrounded by hypointense areas. Histologically, all cases presented granulomas with central necrosis, a polymorphic infiltrate, few giant cells and peripheral fibrosis. The diagnostic yield with conventional microbiologic techniques was poor, whereas a Brucella PCR-assay of a tissue or pus sample was positive in all six cases in which it was performed. Conservative therapy with antibiotics, either alone or combined with percutaneous drainage, failed in all cases, so that in this type of lesion, the treatment of choice should be medical-surgical, in order to guarantee excision of the central calcium nucleus responsible for the persistence of the infection. PMID- 11929687 TI - Changes in the prevalence of hepatitis C virus (HCV) genotype 4 in Calabria, Southern Italy. AB - The aim of this study was to assess changes in the prevalence of hepatitis C virus (HCV) genotypes, focusing on genotype 4, by surveying population of chronic hepatitis C patients within an area of Southern Italy. HCV-RNA was detected in serum using two commercial hepatitis C RNA PCR assays (Amplicor Roche Diagnostic System, and AmpliSensor HCV, Nuclear Laser Medicine). PCR products were analyzed for genotyping using a reverse hybridization of the amplified product by a line probe assay (INNO LIPA, Innogenetics). In our Institution we have previously observed, in a period of 18 months (January 1997-May 1998) an initial increase of the genotype 4 which appeared in 3.3% of HCV patients versus a percentage of 1.3%, during 1996. Later data obtained from 702 HCV-RNA positive patients, collected from June 1998 until December 1999 indicated a 3.7% of genotype 4. This percentage increased until to 4.7% in the most recent period studied (January 2000-February 2001). Drug addiction, blood transfusion and sporadically acquired infections represented the most frequent risk factors. In the Calabria region, genotype 1b, the most prevalent isolate (53.3%) and genotype 2a/2c (26.2%) were associated with older age, confirming our previous study. Genotype 4 was the fifth most prevalent genotype observed, just after 3a and 1a subtypes. Spread of genotype 4 in Calabria region is mostly associated to older age when compared to genotype 3a and 1a, but is statistically associated with a younger group of patients when compared with genotype 1b. In conclusion we demonstrated a fourfold increased prevalence of HCV genotype 4 during the last 5 years. PMID- 11929688 TI - Comparison of line probe assay and DNA sequencing of 5' untranslated region for genotyping hepatitis C virus: description of novel line probe patterns. AB - We compared a commercial line probe assay (INNO-LiPA HCV II, Innogenetics, N.V., Ghent, Belgium, distributed by Bayer Diagnostics) to an in-house 5' untranslated region direct DNA sequencing method for genotyping hepatitis C virus (HCV). Initial evaluation demonstrated that the INNO-LiPA HCV II assay and sequencing assay assigned the same genotype for 110/132 (83.3%) patient specimens (98 subtype and 12 genotype only identifications). Following the initial evaluation, the INNO-LiPA HCV II assay was used routinely to genotype HCV from patient specimens submitted to our laboratory for genotyping (n = 1,739). During this second part of the study, novel line probe patterns have been noted and interpreted using the in-house direct sequencing assay. Reactivity at bands 1, 2, 3, 4, 5 and 8 (n = 4) or 1, 2, 3, 4, 6 and 7 (n = 2) represented HCV genotype 1. Reactivity at bands 1, 2, 5 and 9 (n = 1) represented HCV genotype 2. Reactivity at bands 1, 2, 5, 9 and 16 (n = 1) represented HCV genotype 4. Reactivity at bands 1, 2, 5, 9, 10, 11 (weak band) and 12 (n = 118) most likely represented HCV genotype 2b. This information should be of use to INNO-LiPA HCV II assay users. PMID- 11929689 TI - Comparison of Albicans ID2 agar plate with the germ tube for presumptive identification of Candida albicans. AB - Albicans ID2 (bioMerieux, France) is a commercially available chromogenic medium that allows rapid and specific macroscopic identification of Candida albicans and facilitates the differentiation of species in mixed cultures. We compared it with the standard method for the identification of yeast species, the germ tube test (GT). This study involved 423 clinical isolates, including 163 C. albicans and 260 non-albicans yeasts. Sensitivity of Albicans ID2 agar plates regarding the identification of C. albicans were 98.2% after 48 h of incubation and specificity of 96.6%. This method using rapid enzymatic method shows the same similar sensitivity than the GT test The false negative rate (1.8%) for the GT test is consistent with that previously reported. None tests discriminated between C. albicans and C. dubliniensis isolates. PMID- 11929690 TI - Trypanosoma cruzi: isolation of an immunodominant peptide of TESA (Trypomastigote Excreted-Secreted Antigens) by gene cloning. AB - Trypomastigote forms of Trypanosoma cruzi excrete-secrete several molecules, which are immunodominant during the human infection. This complex antigenic mixture termed TESA (Trypomastigote Excreted-Secreted Antigens) presents a 150 160 kDa band that shows excellent specificity and sensitivity in Chagas' disease diagnosis by immunoblotting. Here we describe the isolation and the antigenic characterization of a recombinant peptide (TESA-1) containing a 10 kDa T. cruzi peptide that belongs to the 150-160 kDa TESA fraction. The clone was isolated by screening a T. cruzi genomic expression library with chagasic antibodies reactive to the 150-160 kDa band of TESA immunoblots. After expression, the recombinant peptide TESA-1 was purified and used to immunize rabbits. Anti-TESA-1 immunesera specifically recognized the 150-160 kDa fraction of TESA-blots from eight different T. cruzi strains. The TESA-1 peptide reacted with 82.2% of chagasic patient sera by immunoblotting, showing that it harbors most of the antigenic epitopes that account for the high reactivity of the 150-160 kDa band of TESA. PMID- 11929691 TI - Prevalence of extended spectrum beta-lactamase (ESBL)-producing clinical isolates in the Asia-Pacific region and South Africa: regional results from SENTRY Antimicrobial Surveillance Program (1998-99). AB - The frequency of occurrence of ESBL-producing clinical strains varies widely in distinct geographic areas. The objective of this study was to describe the frequency of occurrence, the preferred substrate, and the co-resistance patterns of the ESBL-producing isolates collected from the Asia-Pacific region and South Africa through the SENTRY Antimicrobial Surveillance Program between January 1998 and December 1999. A total of 1,377 Escherichia coli, 678 Klebsiella pneumoniae, and 138 Proteus mirabilis isolates were collected from diverse body sites. Using NCCLS criteria, 139 E. coli (10.1%), 171 K. pneumoniae (25.2%), and 2 P. mirabilis (1.4%) had presumptive ESBL phenotypes; 100, 146 and 1 strain respectively were confirmed to be ESBL producers on clavulanate enhancement testing. The frequency of occurrence of confirmed ESBL-producing E. coli by the medical centers varied from 0-1% for centers located in Australia to 13-35% for mainland Chinese centers. The higher prevalence rates (>20%) of ESBL K. pneumoniae phenotypes were observed in all mainland Chinese centers, one Japanese and one Taiwanese center, and in the Philippine, South African, Singaporean and medical centers. The spread of the presumptive ESBL phenotype to the Enterobacter species was observed in nine medical centers. Overall, ceftriaxone and aztreonam were the best substrates for the detection of the ESBL phenotype between both E. coli isolates and K. pneumoniae ESBL phenotypes; however, there was significant variation between countries in substrate preference. Co-resistances to gentamicin, tobramycin, tetracycline, and trimethoprim-sulfamethoxazole were common throughout isolates collected from most medical centers. Ciprofloxacin resistance rates were very high among isolates collected from Hong Kong, mainland China, Singapore, and the Philippines. The best coverage against ESBL-producing isolates was obtained with imipenem (0% resistance), followed by amikacin (6% resistance). PMID- 11929692 TI - Molecular epidemiology and antimicrobial susceptibility of methicillin-resistant Staphylococcus aureus in Taiwan. AB - Methicillin-resistant Staphylococcus aureus (MRSA) has become an important nosocomial pathogen and has increased frequency in the past 20 years. It is highly transmissible between wards, hospitals, and cities. In most major hospitals in Taiwan, MRSA accounts for more than 60% of the S. aureus isolates. Whether there is a predominant strain, which is spread over the whole of Taiwan has not yet been studied. We collected 208 sequential clinical isolates of MRSA from 22 hospitals: seven in northern Taiwan, seven in western Taiwan, five in southern Taiwan, and three in eastern Taiwan during a three-month period in 1998. Using pulsed-field gel electrophoresis, 112 MRSA isolates were shown to belong to one major type,--type C, and this type was shown to have spread widely across all of Taiwan. Ninety-six isolates belonged to 20 other minor types. Most MRSA isolates of this major type were multi-drug resistant and only susceptible to vancomycin and rifampin. We concluded that the high prevalence of MRSA in Taiwan was partly due to the spreading of a predominant strain and most of them were multi-drug resistant. This might imply that more effort should be made to control the spread of MRSA in Taiwan. PMID- 11929693 TI - Simultaneous detection of the mecA and ileS-2 genes in coagulase-negative staphylococci isolated from Brazilian hospitals by multiplex PCR. AB - Coagulase-negative Staphylococcus spp. (CNS) has been associated with primary bloodstream infections and implanted medical devices. Its importance is increasing due to the acquisition of resistance to oxacillin (Oxa) and, recently, resistance to mupirocin (Mup). Mupirocin, a topical antimicrobial, has been used in the prevention of staphylococci catheter colonization. Susceptibility to Oxa and Mup was analyzed by different testing methods in clinical CNS isolates. Among 112 CNS strains, 69 (61.6%) were Oxa(R) by the disk diffusion (DD) method and 72 (64.2%) grew on the oxacillin agar screen plate. S. epidermidis and S. haemolyticus presented high rates of oxacillin resistance, 75.4% and 96.1%, respectively. Twenty four (21.4%) strains were Mup(R) by the DD test and 21 of them (87.5%) were identified as S. epidermidis. The detection of the mecA and ileS-2 genes, determined by multiplex-PCR, showed that 72 (64.2%) CNS strains possessed the mecA gene, while 16 (14.3%) possessed the ileS-2 gene. Fifteen of these strains presented the two resistance genes simultaneously. The isolates containing the ileS-2 gene presented a minimum inhibitory concentration (MIC) >1024 microg/mL in the E-test, while low-level mupirocin resistance (MICs of 12 16 microg/mL) was observed in those strains without ileS-2. The resistances to high and low levels of mupirocin could not be distinguished when the DD test was used. The analysis of the Mup(R) S. epidermidis strains by Pulsed Field Gel Electrophoresis showed that 17 (80.9%) strains belonged to one of two patterns (A and B), which have been shown to be prevalent in hospitals in Rio de Janeiro. This report showed that the PCR method for detection of oxacillin and mupirocin resistance in CNS is necessary to determine accurate rates of these resistance, and will can help in the staphylococcal infections prevention and control policies in Brazil. PMID- 11929694 TI - Ciprofloxacin as broad-spectrum empiric therapy--are fluoroquinolones still viable monotherapeutic agents compared with beta-lactams: data from the MYSTIC Program (US)? AB - Antimicrobial susceptibility data from United States medical centers participating in the Meropenem Yearly Susceptibility Test Information Collection (MYSTIC) surveillance program were analyzed. These results indicate that ciprofloxacin resistance in commonly encountered nosocomial pathogens has reached a level where fluoroquinolones may no longer provide assured empiric monotherapy for serious infections nor provide a spectrum as a co-drug equivalent to aminoglycosides. Local susceptibility testing data must be assessed before fluoroquinolone empiric therapy can be used with confidence. PMID- 11929695 TI - A new integron carrying VIM-2 metallo-beta-lactamase gene cassette in a Serratia marcescens isolate. AB - Serratia marcescens is an important nosocomial pathogen which is often resistant to multiple antimicrobial agents. An imipenem-resistant S. marcescens isolate from a urine specimen was found to carry a bla(VIM-2) gene cassette on a class 1 integron. This finding indicates that bla(VIM-2) is presently spreading even to Serratia spp. in Korea, which could compromise the usefulness of carbapenem in the treatment of multi-resistant Gram-negative bacilli infections. Clinical laboratory should be able to detect the VIM-2-producing isolates with even low carbapenem MIC. PMID- 11929696 TI - Basis of FLT as a cell proliferation marker: comparative uptake studies with [3H]thymidine and [3H]arabinothymidine, and cell-analysis in 22 asynchronously growing tumor cell lines. AB - The usefulness of radiolabeled 3'-fluoro-3'-deoxythymidine (FLT), a thymidine derivative with affinity to cytoplasmic thymidine kinase 1 (TK1), as a tumor proliferation marker was evaluated using [3H]FLT and 22 cultured tumor cell lines. Asynchronously growing tumor cells were used for studies to mimic in vivo status of tumors. FLT uptake in each cell line was compared with [3H]thymidine ([3H]Thd) uptake and %S-phase fraction, both known as acceptable markers of proliferation. Uptake of the mitochondrial TK2 specific substrate [3H]arabinothymidine ([3H]AraT) was studied as a reference. Metabolic fate of FLT in tumor cells was also analyzed to elucidate the retention mechanism of FLT. [3H]FLT uptake was mildly correlated with the %S-phase fraction (r=0.76, p<0.0001) and correlated better with [3H]Thd uptake (r=0.88, p<0.0001). In contrast, the TK(2) specific substrate, [3H]AraT, was not significantly correlated with the %S-phase fraction (r=0.19, p=0.39), although it showed some correlation with the [3H]Thd uptake (r=0.47, p<0.05). Over 90% of radioactivity of [3H]Thd was found in the DNA fraction after 60 minutes incubation. In contrast, most of the radioactivity of [3H]FLT was found in the acid-soluble fraction (95%). [3H]FLT incorporation into the DNA fraction was negligible (0.2%). The [3H]AraT was mainly distributed in the acid-soluble fraction (70%) and the DNA fraction (20%). From our results, we concluded that FLT uptake in tumor cells reflects tumor cell proliferation. However, much more convincing validation is needed to clarify the difference between FLT and true substrates for DNA synthesis, like thymidine. PMID- 11929697 TI - Synthesis and biodistribution of a Lipophilic 64Cu-labeled monocationic Copper(II) complex. AB - The lipophilic, monocationic copper(II) complex of the diiminedioxime ligand 2,10 di-n-butyl-3,9-dimethyl-1,4,8,11-tetraazaundeca-1,3,8,10-tetraen-1,11-dione dioxime was synthesized and labeled with 64Cu. The biological properties of the 64Cu-labeled complex were measured in vivo and in vitro. In vivo, the complex shows uptake by the heart similar to that of 99mTc-tetrofosmin. In vitro, its uptake by multidrug resistant and non-resistant MES-SA tumor cells parallels that of 99mTc-MIBI, a well-characterized marker of multidrug resistance. These results suggest that this class of copper complexes may form the basis for the development of a 64Cu PET radiopharmaceutical for the functional imaging of multidrug resistance and/or myocardial perfusion. PMID- 11929698 TI - (+)-[76Br]A-69024: a non-benzazepine radioligand for studies of dopamine D1 receptors using PET. AB - (+)-[76Br]A-69024 is a specific and enantioselective dopamine D1 receptor radioligand. The Bmax of (+)-[76Br]A-69024 measured in vitro on rat striatum membranes was 320 +/- 25 fmoles/mg protein with an apparent dissociation constant of Kd = 0.6 +/- 0.1 nM. The inactive enantiomer, (-)-[76Br]A-69024, displayed no affinity in the same assay. In vivo, the biodistribution (+)-[76Br]A-69024 in rats showed a rapid and high uptake in the striatum (1% ID/g), followed by a slow wash out. The striatum/cerebellum concentration ratio (index of specific binding) reached a maximum value of 10 at 60 minutes post injection. A tissue to cerebellum ratio of 2.8 and 1.5 was also observed for frontal and posterior cortex respectively. With the pharmacologically inactive enantiomer, (-)-[76Br]A 69024, the brain uptake was determined to be non specific since a striatum/cerebellum ratio of approximately 1 was observed throughout the time course of the experiment. The selectivity of (+)-[76Br]A-69024 uptake was demonstrated in competition experiments. The specific uptake in the striatum and cortical regions was completely prevented after administration of the D1 antagonist SCH 23390. Pre-treatment of rats with unlabelled (+)A-69024 also displayed the same regional inhibition of (+)-[76Br]A-69024 uptake. Pre administration of rats with spiperone (D2) and ketanserin (5-HT2/5-HT2C) showed no inhibitory effect on (+)-[76Br]A-69024 uptake in any brain region. Using (+) [76Br]A-69024, PET study in baboon demonstrated a preferential accumulation of the radioactivity in the striatum, frontal and posterior cortex which was displaced to the level of the cerebellum by SCH 23390. These results suggest that (+)-[76Br]A-69024 may deserve further investigation as a potential radioligand for studying striatal and cortical dopamine D1 receptors using PET. PMID- 11929699 TI - In vitro uptake of [1-14C]Octanoate in brain slices of rats: basic studies for assessing [1-11C]Octanoate as a PET tracer of glial functions. AB - To clarify the contribution of glial cells to octanoate uptake into the brain, we determined the effects of fluoroacetate, a selective inhibitor of glial metabolism, on in vitro brain uptake of [1-14C]octanoate, using rat brain slices. The [1-14C]octanoate uptake significantly decreased, depending on the concentration of fluoroacetate (p = 0.001). The [1-14C]octanoate uptakes at 5 mM (0.23 +/- 0.05% uptake/mg slice) and 25 mM fluoroacetate (0.12 +/- 0.01% uptake/mg slice) were significantly lower than that at control (0.29 +/- 0.02% uptake/mg slice, p < 0.05 and p < 0.001, respectively). The results demonstrate the contribution of glial cells to octanoate uptake into the brain. The potential of [1-11C]octanoate as a PET tracer for studying glial functions is suggested. PMID- 11929700 TI - Positron emission tomography and ex vivo and in vitro autoradiography studies on dopamine D2-like receptor degeneration in the quinolinic acid-lesioned rat striatum: comparison of [11C]raclopride, [11C]nemonapride and [11C]N methylspiperone. AB - With [11C]raclopride,[11C]nemonapride and [11C]N-methylspiperone, degeneration of dopamine D2-like receptors in the unilaterally quinolinic acid-lesioned rats was evaluated by positron emission tomography (PET) and ex vivo and in vitro autoradiography. PET showed a decreased uptake of [11C]raclopride in the lesioned striatum, but an increased uptake of [11C]nemonapride and [11C]N-methylspiperone despite a decreased binding in vitro. Ex vivo autoradiography showed an increased accumulation of the three ligands in the cortical region overlying the injured striatum, probably enlarging PET signals. PET has the limited potential for evaluating the receptor degeneration in the present animal model. PMID- 11929701 TI - PET evaluation of a tetracyclic, atypical antidepressant, [N-methyl 11C]mianserin, in the living porcine brain. AB - We synthesized [N-methyl-11C]mianserin by alkylation of N-desmethyl mianserin with [11C]methyl iodide followed by HPLC purification. We used PET for determining the regional cerebral pharmacokinetics of the radiotracer in anesthetized swine. [N-methyl-11C]Mianserin entered most brain regions readily (range of K1 values: 0.66-1.13), reaching highest levels in the basal ganglia and thalamus. The binding potential of [N-methyl-11C]mianserin was relatively low (range: 0.07-0.50), but regional differences were nonetheless observed, with highest values in the temporal cortex and lowest values in the brainstem. These PET findings, which are the first ones for a tetracyclic, antidepressant drug, show that [N-methyl-11C]mianserin has only a limited degree of regional specificity of binding in the living brain. PMID- 11929702 TI - Stabilization of neurotensin analogues: effect on peptide catabolism, biodistribution and tumor binding. AB - Neurotensin (NT) receptors in pancreatic and other neuroendocrine tumors are promising targets for imaging and therapeutic purposes. Here, we report on the effect of distinct changes in the peptide chain on catabolism in vitro for five radiolabeled [99mTc] neurotensin analogues having high affinity for neurotensin receptors. Substitution of NT(1-7) by (NalphaHis)Ac--the Tc-binding moiety- combined with a reduced bond 8-9 (CH2NH), N-methylation of peptide bonds or replacement of Ile(12) by tertiary leucin (Tle) led to peptide stabilization of various degrees. Biodistribution studies in nude mice bearing HT29 xenografts showed higher tumor uptake with more stable peptides, yielding high tumor to blood ratios of up to 70. PMID- 11929703 TI - Synthesis, characterization and biodistribution of bisphosphonates Sm-153 complexes: correlation with molecular modeling interaction studies. AB - Bisphosphonates (BPs) are characterized by a P-C-P backbone structure and two phosphonic acid groups bonded to the same carbon, and are established as osteoclast-mediated bone resorption inhibitors. The nature of the groups attached to the central carbon atom are responsible in determining the potency of bisphosphonates as anti-resorption drugs. However, it is not yet clear the exact relationship between their molecular structure and pharmacologic activities. In this study, molecular geometries of pamidronate, alendronate and neridronate, differing only in the length of the aliphatic chains, were predicted by molecular mechanics and their interactions with hydroxyapatite, the main bone mineral component, were examined. We report the synthesis and radiochemical characterization of 153Sm complexes with pamidronate, alendronate and neridronate. Hydroxyapatite binding and biodistribution studies of these complexes have shown a good correlation with the theoretical molecular modeling interaction studies. So, it is possible to conclude that computational chemistry techniques are a good approach to evaluate specific interactions and may play a relevant role in determining the relative ability of BPs to mineral bone, and open new perspectives to the design of new BPs with increased pharmacological activity. These techniques could be extended to BPs as ligands to carrier radioactive metals, aiming for new bone therapeutic radiopharmaceuticals. PMID- 11929704 TI - Detecting parathyroid adenoma using technetium-99m tetrofosmin: comparison with P glycoprotein and multidrug resistance related protein expression--a preliminary report. AB - The aim of this study was to investigate the relationships among technetium-99m tetrofosmin (Tc-TF) accumulation in parathyroid adenoma and the expression of P glycoprotein (Pgp) or multidrug resistance related protein (MRP). Before operation, 33 patients with parathyroid adenomas (larger than 1.5 gm) were studied with parathyroid scintigraphy 10 minutes and 2 hours after intravenous injection of Tc-TF before operation. Immunohistochemical analyses (IHA) were performed on multiple nonconsecutive sections of operative parathyroid specimens to detect Pgp or MRP expression. According to the results of IHA, the 33 parathyroid adenomas were separated into four groups: (1) 2 adenomas with both positive Pgp and positive MRP expression, (2) 1 adenomas with positive Pgp but negative MRP expression, (3) 2 adenomas with negative Pgp but positive MRP expression, and (4) 28 adenomas with both negative Pgp and negative MRP expression. All of 28 adenomas in the group 4 could be detected by Tc-TF parathyroid imaging. All of 5 adenomas in the groups 1 to 3 could not be detected by TcTF parathyroid imaging (p < 0.05). Not only the size of parathyroid adenomas, but also significant Pgp or MRP expression limited the sensitivity of Tc-TF parathyroid imaging to localize parathyroid adenomas before operation. PMID- 11929705 TI - A simplified and improved synthesis of [11C]phosgene with iron and iron (III) oxide. AB - [11C]Phosgene ([11C]COCl2), a useful precursor for labeling several radiopharmaceuticals, is generally produced by catalytic oxidation of [11C]carbon tetrachloride over Fe granules, although in low yields or with poor reproducibility. In order to develop am improved synthesis of [11C]phosgene, two oxidizing agents, Fe2O3 and CuO, were examined. The yield of [11C]phosgene was significantly increased using Fe2O3 powder mixed with Fe granules, while the use of CuO alone, or CuO powder mixed with Fe granules resulted in an insignificant yield. The yield and specific activity of S- (-) [11C]CGP-12177 synthesized using Fe2O3 powder mixed with Fe granules were markedly higher than those synthesized by the previous methods using Fe granules alone or Fe granules mixed with Fe powder. Thus, in the present study, we developed a simple and practical method for the synthesis of [11C]phosgene, which provided an improved yield of S- (-) [11C]CGP-12177. PMID- 11929706 TI - Supercritical CO2 fluid radiochromatography system used to purify [11C]toluene for PET. AB - Abuse of inhalants in today's society has become such a widespread problem among today's adolescents that in many parts of the world their use exceeds that of many other illicit drugs or alcohol. Even so, little is known how such inhalants affect brain function to an extent that can lead to an abuse liability. While methodologies exist for radiolabeling certain inhalants of interest with short lived positron emitting radioisotopes that would allow their investigation in human subjects using positron emission tomography (PET), the purification methodologies necessary to separate these volatile substances from the organic starting materials have not been developed. We've adapted supercritical fluid technology to this specific PET application by building a preparative-scale supercritical CO2 fluid radiochromatograph, and applied it to the purification of [11C]toluene. We've demonstrated that [11C]toluene can be separated from the starting materials using a conventional C18 HPLC column and pure supercritical CO2 fluid as the mobile phase operating at 2000 psi and 40 degrees C. We've also shown that the purified radiotracer can be quantitatively captured on Tenax GR, a solid support material, as it exits the supercritical fluid stream, thus allowing for later desorption into a 1.5% cyclodextrin solution that is suitable for human injection, or into a breathing tube for direct inhalation. PMID- 11929707 TI - Improved method for the quality assurance of [C-11]choline. AB - [C-11]choline is a very promising radiomarker for the diagnosis of various human tumors using Positron Emission Tomography (PET). The existing quality control process for [C-11]choline is complicated and combines two HPLC methods with limited separation and sensitivity which prevent the accurate determination of the specific activity. We have developed a new efficient single HPLC method for the detection of choline chloride and dimethylaminoethanol with high resolution and sensitivity using cation-exchange chromatography. PMID- 11929708 TI - Is enteral administration of fluorine-18-fluorodeoxyglucose (F-18 FDG) a palatable alternative to IV injection? Pre-clinical evaluation in normal rodents. AB - To establish effective methods of enteral 2-[18F]-fluoro-2-deoxy-D-glucose (FDG) administration, the efficiency of FDG absorption in the gastrointestinal tracts following enteral administrations was evaluated using the FDG biodistribution in normal rodents, in combination with various fasting conditions and FDG diluents. The blood FDG curve using hypotonic solution showed a rapid increase, while that in iso- and hypertonic groups showed slow rises. Brain FDG uptake had a close positive correlation with blood AUC (area under curve) and an inverse relationship with the stomach contents. PMID- 11929709 TI - Dendritic cell subtypes in autoimmune liver diseases; decreased expression of HLA DR and CD123 on type 2 dendritic cells. AB - In order to dissect the role of different subsets of antigen-presenting dendritic cell (DC) in autoimmune liver diseases, we analyzed the frequencies and phenotypes of DC1 and DC2 from patients with primary biliary cirrhosis (PBC) and autoimmune hepatitis (AIH). The numbers of DC, DC1 and DC2 in a fixed amount of peripheral blood (20 ml) from 14 patients with PBC (therapy [-]: 5, therapy [+]: 9), seven patients with AIH (therapy [-]): 4, therapy [+]: 3), nine patients with chronic hepatitis due to hepatitis C virus and 12 healthy subjects were estimated using three-color flow cytometry. The mean fluorescence intensities of HLA DR, CD11c and CD123 on DC subsets were evaluated from the flow cytometric profiles. The numbers of DC1 and DC2 did not differ significantly among patients and controls. The expression of HLA DR was lower on DC1 from patients with AIH, but not on PBC. However, the levels of HLA DR and CD 123 on DC2 were significantly decreased in patients with PBC (therapy [-]) and AIH (therapy [-]) compared with healthy subjects (P<0.05). Defective phenotype of DC2 in both AIH and PBC may have a relevance to the breakdown of tolerance to self antigen in these autoimmune diseases. Functional study of DC2 in autoimmune liver diseases is warranted. PMID- 11929710 TI - Long-term prognosis of primary biliary cirrhosis (PBC) in Japan and analysis of the factors of stage progression in asymptomatic PBC (a-PBC). AB - Objective: Based on data from a national survey of primary biliary cirrhosis (PBC), the pathology and prognosis of PBC in Japan were clarified. In particular, we tried to perform multivariate analysis of factors useful in determining prognosis of asymptomatic PBC (a-PBC). Methods: The survey was performed 10 times. Responses from 3778 of 4361 registered patients (416 institutions) were investigated (survey period: January 1968-December 1998). At the time of diagnosis, patients were classified as a-PBC or symptomatic PBC (s1-PBC; pruritus only, s2-PBC; jaundice and serum bilirubin level above 2 mg/dl). The survival rate was obtained by the Kaplan-Meier method. Logistic regression analysis was used in multivariate analysis of prognostic factors of a-PBC. Results: There were no significant differences in clinical findings from those in previous reports. The 5-year survival rates of patients with a-PBC, s1-PBC, and s2-PBC at the time of diagnosis were 97, 88, and 53%, respectively. Patients with a-PBC at the time of diagnosis were divided into groups: those in whom the disease progressed to s2 PBC (8%) and did not progress to s2-PBC (92%) at the final examination, and the prognosis was compared between groups. The prognosis was significantly poorer in the s2-PBC progression group. As a result of multivariate analysis for prediction of prognosis, levels at diagnosis of total serum bilirubin (T-Bil), albumin (Alb), total cholesterol (T-Cho), histological stage, and presence or absence of ursodeoxycholic acid (UDCA) administration were selected as significant factors (P<0.00001). Conclusion: Serum T-Bil, Alb, T-Cho, and histological stage at the time of diagnosis and presence or absence of UDCA administration were considered useful early prognostic indicators in patients diagnosed as having a-PBC whose prognosis may deteriorate with progression to s2-PBC. PMID- 11929711 TI - Serum and tissue PIVKA-II expression reflect the biological malignant potential of small hepatocellular carcinoma. AB - A sensitive method for measuring the serum level of protein-induced by vitamin K absence or antagonist II (PIVKA-II) has become so widely available that it is now used for the clinical diagnosis of small hepatocellular carcinoma (HCC). It is known that serum PIVKA-II can be a prognostic indicator for HCC, but there have been no detailed investigations concerning the tissue expression of PIVKA-II. The present study assessed the relationship between serum or tissue PIVKA-II and the biological malignant potential of HCC. The subjects were 25 patients with histologically confirmed HCC, that were solitary and 3 cm or less in diameter. Tissue PIVKA-II was detected by immunostaining using MU-3 as the primary antibody. The biological malignant potential of the tumors was evaluated on the basis of the Ki-67 labeling index of HCC cells and the tumor arterial vascularity assesed by angiography and CO(2) enhanced ultrasonography. The recurrence-free period after treatment was also evaluated. Among the 25 patients, eight were positive for tissue PIVKA-II. Serum PIVKA-II levels were significantly higher in the tissue PIVKA-II-positive patients compared with the negative patients, but serum and tissue PIVKA-II expressions were not consistently parallel. Tumor cell proliferation was closely correlated with the tissue PIVKA-II expression, while the recurrence-free period was correlated with the serum PIVKA-II level. Tumor arterial vascularity showed a strong correlation with the expression of both serum and tissue PIVKA-II. In conclusion, serum and tissue PIVKA-II expression reflect the biological malignant potential of HCC and thus may be useful indicators for the prognosis of small HCC. PMID- 11929712 TI - Gene expression of interleukin-12 receptor beta2 chain and Th1 population of peripheral blood mononuclear cells in chronic hepatitis C. AB - Gene expression of interleukin 12-receptor beta2 chain mRNA in peripheral blood mononuclear cells (PBMCs) was examined in patients with chronic hepatitis C (n=7) and in healthy control subjects (n=6) by semi-quantitative RT-PCR. The level of interleukin 12-receptor beta2 chain mRNA was higher in patients with chronic hepatitis C than in healthy subjects (P=0.032). The level of interleukin 12 receptor beta2 chain mRNA had a weak correlation with the ratio of Th1 to Th2 populations (r=0.714, P=0.020). There was a tendency for the level of interleukin 12-receptor beta2 mRNA to increase both in chronic hepatitis C (P=0.109) and in healthy volunteers (P=0.144) after the incubation of PBMCs with interferon-alpha in vitro. During interferon-alpha administration to the patients with chronic hepatitis C, the level of interleukin 12-receptor beta2 chain mRNA in PBMCs was increased in all four cases. Although this is a preliminary study with a small sample size, our results suggest that the level of interleukin 12-receptor beta2 chain mRNA is higher than normal in patients with chronic hepatitis C and can be further enhanced by interferon therapy. PMID- 11929714 TI - The clinical significance of GBV-C/HGV exposure in C-viral chronic liver disease and blood donors. AB - Aims: The relationship between the hepatitis G virus (HGV) RNA-positive state or the HGV anti-E2 antibody-positive state, and various clinical parameters among patients with C-viral chronic liver disease and blood donors, was examined. Patients and methods: The subjects consisted of 166 patients with hepatitis C virus (HCV) RNA-positive liver disease, and 157 blood donors. Serum samples were tested for the presence of HGV RNA by the polymerase chain reaction method. The HGV E2 antibody level was measured with an enzyme-linked immunosorbent assay kit. Results: HGV RNA was detected in 1.3% of the blood donors, and HGV E2 antibodies were detected in the sera of 2.5% of the blood donors. The detection rate of HGV RNA and HGV E2 antibodies among the patients with C-viral liver disease was 4.8 and 28.3%, respectively. The detection rates of HGV RNA and HGV E2 antibody among those in each F stage were 0 and 25.0% among those in the F1 stage, 2.6 and 34.2% among those in F2, 11.1 and 27.8% among those in F3, 12.5 and 29.2% among those in F4, and 11.1 and 27.8% among those with hepatocellular carcinoma (HCC). The detection rate of HGV RNA increased with the progression of the F stage and HCC, however, the detection rate of HGV E2 antibody among the four F stages and HCC did not significantly differ. In addition, the clinical parameters and background of those who did or did not have HGV E2 antibodies or HGV RNA, did not significantly differ. Conclusion: HGV exposure may promote the progression of liver fibrosis (F stage) in C-viral liver diseases. PMID- 11929713 TI - Aromatase-deficient (ArKO) mice are retrieved from severe hepatic steatosis by peroxisome proliferator administration. AB - Tamoxifen is a potent antagonist of estrogen, and hepatic steatosis is a frequent complication in adjuvant tamoxifen for breast cancer. Recently, aromatase deficient (ArKO, Ar-/-) mice lacking intrinsic estrogen was developed and the molecular mechanism involved in progression of massive hepatic steatosis in estrogen-deficiency was elucidated; impairment in hepatic fatty acid beta oxidation of peroxisomes, microsomes and mitochondria. This impairment is latent, but is potentially serious, because hepatic energy supply depends greatly on fatty acid beta-oxidation. Therefore in the present study, we tried to conquer impaired hepatic fatty acid beta-oxidation by administrating bezafibrate, a potent peroxisome proliferator, to Ar-/- mice through activating fatty acid beta oxidation via the peroxisome proliferator activated receptor-alpha mediated signaling pathway. Northern blot analysis of Ar-/- mice liver revealed a significant restoration of mRNA expression of very long fatty acyl-CoA synthetase in peroxisome, peroxisomal fatty acyl-CoA oxidase, and medium-chain acyl-CoA dehydrogenase in mitochondria, essential enzymes in fatty acid beta-oxidation by administration of bezafibrate. Severe hepatic steatosis observed in Ar-/- mice regressed dramatically. Consistent findings were obtained in the in vitro assays of fatty acid beta-oxidation activity. These findings demonstrate that bezafibrate is capable of restoring impaired fatty acid beta-oxidation in vivo via the peroxisome proliferator-activated receptor-alpha mediated signaling pathway and is potent enough to regress severe hepatic steatosis in mice deficient in intrinsic estrogen. PMID- 11929715 TI - Hepatitis C virus core protein binds to a C-terminal region of NS5B RNA polymerase. AB - Hepatitis C virus (HCV) NS5B has been shown to exhibit RNA-dependent RNA polymerase activity for its viral RNA replication. In this study, we demonstrated the formation of a complex between NS5B and the core protein (NS5B-core protein complex) in mammalian cells, as determined by indirect immunofluorescence and immunoprecipitation analyses. The localization of the core protein was observed to change to the same locus in ER as NS5B locates by its coexpression with NS5B, indicating that the localization of the core protein is determined by NS5B. The truncated NS5B molecule lacking the C-terminal region did not form a complex with the core protein, suggesting that the C-terminal region of NS5B is essential for its interaction with the core protein. Moreover, the change in NS5B localization because of C-terminal deletion indicates that this region includes a certain signal for NS5B retention in ER. PMID- 11929716 TI - Septic endophthalmitis and meningitis associated with Klebsiella pneumoniae liver abscess. AB - We report a female case of septic endophthalmitis and meningitis associated with Klebsiella pneumoniae liver abscess which was thought to be caused by duodeno biliary reflux related to choledochoduodenostomy. We treated this patient by ultrasonography-guided percutaneous abscess drainage and intravenous administration of third generation antibiotics. However, the visual function of her left eye was eventually lost. Reports of liver abscess with metastatic lesions are rare, and our experience suggests that more physicians should be alert to septic metastatic lesions such as K. pneumoniae liver abscess or bacteremia with complaints of ocular or central nervous system symptoms. PMID- 11929717 TI - CEA producing primary hepatic carcinoid. AB - Imaging studies of a hepatic tumor in a 53-year-old woman with elevated serum levels of neuron-specific enolase (NSE), carcinoembryonic antigen (CEA) and 5 hydroxyindole acetic acid (5HIAA) revealed a hypervascular tumor in the right lobe. Grossly, the brownish tumor was measured 13.5x12 cm with four daughter nodules. Microscopically, the majority of these columnar and round tumor cells had ribbon-or rosette-like patterns with the expression of neuroendocrine marker proteins, such as Grimelius, NSE, chromogranin A, and synaptophysin, and moderate expression of CEA but without the expression of cytokeratin nos 7,8,14,18,19 and OV-6; the minority had glandular patterns with a strong expression of CEA but without the expression of cytokeratin nos 7,8,14,18,19 and OV-6. Ultrastructurally, most tumor cells contained populations of electron-dense core granules ranging between 100 and 200 nm in diameter. After hepatectomy, serum CEA, NSE, and 5HIAA reverted to normal ranges and persisted for 19 months. These findings suggested that the diagnosis of primary hepatic carcinoid was tenable and that the tumor might derive from hepatic stem cells which acquired the additional nature of producing CEA without cytokeratins characteristic of hepatocytes or bile duct cells. Some molecular based approaches have attributed unique biological behavior and histogenesis to this carcinoid tumor. PMID- 11929718 TI - Erratum to "Anti-fibrogenic effect of an angiotensin converting enzyme inhibitor on chronic carbon tetrachloride-induced hepatic fibrosis in rats". [Hepatology Research 21/2, 147-158 (2001)]. PMID- 11929719 TI - Erratum to "Th1 and Th2 cytokines differentially regulate the transformation of Kupffer cells into multinucleated giant cells but similarly enhance the Kupfer cell-induced hepatic stellate cell proliferation". [Hepatology Research 20/2, 193 206 (2001)]. PMID- 11929721 TI - Environmental justice: building a unified vision of health and the environment. AB - The assorted and multidimensional concerns that give rise to the issue of environmental justice have proved to be intellectually daunting and highly resistant to positive change. Low-income, people of color, and tribal communities confronting environmental stressors are beset by stressors in both the physical and social environments. For this reason, while the bifurcation of the public health and environmental fields taking place over the past several decades has yielded generally negative impacts in areas of public health, environment, and planning, the consequences for low-income and disadvantaged communities have been especially grievous. This commentary builds on the recent Institute of Medicine workshop titled "Rebuilding the Unity of Health and the Environment: A New Vision of Environmental Health for the 21st Century." The workshop organizers posited that only by thinking about environmental health on multiple levels will it be possible to merge various strategies to protect both the environment and health. In this commentary we examine how such a new vision of uniting public health and the environment can contribute to attaining environmental justice for all populations. PMID- 11929722 TI - Community involvement in the ethical review of genetic research: lessons from American Indian and Alaska Native populations. AB - The National Bioethics Advisory Commission has proposed that regulatory oversight for research with human subjects be extended beyond the protection of individual research participants to include the protection of social groups. To accomplish this, the commission recommends that investigators and ethics review boards a) work directly with community representatives to develop study methods that minimize potential group harms, b) discuss group implications as part of the informed consent process, and c) consider group harms in reporting research results. We examine the utility of these recommendations in the context of research with American Indian and Alaska Native communities. Because much attention has been given to the question of how best to consult with members of these communities in the design and conduct of research, we believe it behooves investigators to consider the lessons to be learned from research involving American Indians and Alaska Natives. After describing several difficulties surrounding the application of the commission's approach to these research contexts, we propose a research agenda to develop best practices for working with local communities in the ethical assessment of epidemiologic and environmental health research. PMID- 11929723 TI - Environmental justice and regional inequality in southern California: implications for future research. AB - Environmental justice offers researchers new insights into the juncture of social inequality and public health and provides a framework for policy discussions on the impact of discrimination on the environmental health of diverse communities in the United States. Yet, causally linking the presence of potentially hazardous facilities or environmental pollution with adverse health effects is difficult, particularly in situations in which diverse populations are exposed to complex chemical mixtures. A community-academic research collaborative in southern California sought to address some of these methodological challenges by conducting environmental justice research that makes use of recent advances in air emissions inventories and air exposure modeling data. Results from several of our studies indicate that communities of color bear a disproportionate burden in the location of treatment, storage, and disposal facilities and Toxic Release Inventory facilities. Longitudinal analysis further suggests that facility siting in communities of color, not market-based "minority move-in," accounts for these disparities. The collaborative also investigated the health risk implications of outdoor air toxics exposures from mobile and stationary sources and found that race plays an explanatory role in predicting cancer risk distributions among populations in the region, even after controlling for other socioeconomic and demographic indicators. Although it is unclear whether study results from southern California can be meaningfully generalized to other regions in the United States, they do have implications for approaching future research in the realm of environmental justice. The authors propose a political economy and social inequality framework to guide future research that could better elucidate the origins of environmental inequality and reasons for its persistence. PMID- 11929724 TI - Community-based participatory research as a tool to advance environmental health sciences. AB - The past two decades have witnessed a rapid proliferation of community-based participatory research (CBPR) projects. CBPR methodology presents an alternative to traditional population-based biomedical research practices by encouraging active and equal partnerships between community members and academic investigators. The National Institute of Environmental Health Sciences (NIEHS), the premier biomedical research facility for environmental health, is a leader in promoting the use of CBPR in instances where community-university partnerships serve to advance our understanding of environmentally related disease. In this article, the authors highlight six key principles of CBPR and describe how these principles are met within specific NIEHS-supported research investigations. These projects demonstrate that community-based participatory research can be an effective tool to enhance our knowledge of the causes and mechanisms of disorders having an environmental etiology, reduce adverse health outcomes through innovative intervention strategies and policy change, and address the environmental health concerns of community residents. PMID- 11929725 TI - Mapping environmental injustices: pitfalls and potential of geographic information systems in assessing environmental health and equity. AB - Geographic Information Systems (GIS) have been used increasingly to map instances of environmental injustice, the disproportionate exposure of certain populations to environmental hazards. Some of the technical and analytic difficulties of mapping environmental injustice are outlined in this article, along with suggestions for using GIS to better assess and predict environmental health and equity. I examine 13 GIS-based environmental equity studies conducted within the past decade and use a study of noxious land use locations in the Bronx, New York, to illustrate and evaluate the differences in two common methods of determining exposure extent and the characteristics of proximate populations. Unresolved issues in mapping environmental equity and health include lack of comprehensive hazards databases; the inadequacy of current exposure indices; the need to develop realistic methodologies for determining the geographic extent of exposure and the characteristics of the affected populations; and the paucity and insufficiency of health assessment data. GIS have great potential to help us understand the spatial relationship between pollution and health. Refinements in exposure indices; the use of dispersion modeling and advanced proximity analysis; the application of neighborhood-scale analysis; and the consideration of other factors such as zoning and planning policies will enable more conclusive findings. The environmental equity studies reviewed in this article found a disproportionate environmental burden based on race and/or income. It is critical now to demonstrate correspondence between environmental burdens and adverse health impacts--to show the disproportionate effects of pollution rather than just the disproportionate distribution of pollution sources. PMID- 11929727 TI - Examining urban brownfields through the public health "macroscope". AB - Efforts to cope with the legacy of our industrial cities--blight, poverty, environmental degradation, ailing communities--have galvanized action across the public and private sectors to move vacant industrial land, also referred to as brownfields, to productive use; to curb sprawling development outside urban areas; and to reinvigorate urban communities. Such efforts, however, may be proceeding without thorough investigations into the environmental health and safety risks associated with industrial brownfields properties and the needs of affected neighborhoods. We describe an approach to characterize vacant and underused industrial and commercial properties in Southeast Baltimore and the health and well being of communities living near these properties. The screening algorithm developed to score and rank properties in Southeast Baltimore (n= 182) showed that these sites are not benign. The historical data revealed a range of hazardous operations, including metal smelting, oil refining, warehousing, and transportation, as well as paints, plastics, and metals manufacturing. The data also identified hazardous substances linked to these properties, including heavy metals, solvents, polycyclic aromatic hydrocarbons, plasticizers, and insecticides, all of which are suspected or recognized toxicants and many of which are persistent in the environment. The health analysis revealed disparities across Southeast Baltimore communities, including excess deaths from respiratory illness (lung cancer, chronic obstructive pulmonary disease, influenza, and pneumonia), total cancers, and a "leading cause of death" index and a spatial and statistical relationship between environmentally degraded brownfields areas and at-risk communities. Brownfields redevelopment is a key component of our national efforts to address environmental justice and health disparities across urban communities and is critical to urban revitalization. Incorporating public health into brownfields-related cleanup and land-use decisions will increase the odds for successful neighborhood redevelopment and long-term public health benefits. PMID- 11929726 TI - Assessment of personal and community-level exposures to particulate matter among children with asthma in Detroit, Michigan, as part of Community Action Against Asthma (CAAA). AB - We report on the research conducted by the Community Action Against Asthma (CAAA) in Detroit, Michigan, to evaluate personal and community-level exposures to particulate matter (PM) among children with asthma living in an urban environment. CAAA is a community-based participatory research collaboration among academia, health agencies, and community-based organizations. CAAA investigates the effects of environmental exposures on the residents of Detroit through a participatory process that engages participants from the affected communities in all aspects of the design and conduct of the research; disseminates the results to all parties involved; and uses the research results to design, in collaboration with all partners, interventions to reduce the identified environmental exposures. The CAAA PM exposure assessment includes four seasonal measurement campaigns each year that are conducted for a 2-week duration each season. In each seasonal measurement period, daily ambient measurements of PM2.5 and PM10 (particulate matter with a mass median aerodynamic diameter less than 2.5 microm and 10 microm, respectively) are collected at two elementary schools in the eastside and southwest communities of Detroit. Concurrently, indoor measurements of PM2.5 and PM10 are made at the schools as well as inside the homes of a subset of 20 children with asthma. Daily personal exposure measurements of PM10 are also collected for these 20 children with asthma. Results from the first five seasonal assessment periods reveal that mean personal PM10 (68.4 39.2 microg/m(3)) and indoor home PM10 (52.2 30.6 microg/m(3)) exposures are significantly greater (p < 0.05) than the outdoor PM10 concentrations (25.8 11.8 microg/m(3)). The same was also found for PM2.5 (indoor PM2.5 = 34.4 21.7 microg/m(3); outdoor PM2.5 = 15.6 8.2 microg/m(3)). In addition, significant differences (p < 0.05) in community-level exposure to both PM10 and PM2.5 are observed between the two Detroit communities (southwest PM10 = 28.9 14.4 microg/m(3)), PM2.5 = 17.0 9.3 microg/m(3); eastside PM10 = 23.8 12.1 microg/m(3), PM2.5 = 15.5 9.0 microg/m(3). The increased levels in the southwest Detroit community are likely due to the proximity to heavy industrial pollutant point sources and interstate motorways. Trace element characterization of filter samples collected over the 2-year period will allow a more complete assessment of the PM components. When combined with other project measures, including concurrent seasonal twice-daily peak expiratory flow and forced expiratory volume at 1 sec and daily asthma symptom and medication dairies for 300 children with asthma living in the two Detroit communities, these data will allow not only investigations into the sources of PM in the Detroit airshed with regard to PM exposure assessment but also the role of air pollutants in exacerbation of childhood asthma. PMID- 11929728 TI - Environmental injustice and the Mississippi hog industry. AB - The recent growth and restructuring of the swine industry in the state of Mississippi has raised various environmental and socioeconomic concerns. We spatially examined the location and attributes of 67 industrial hog operations to determine if African American and low-income communities have a high prevalence of industrial hog operations located near their neighborhoods at the census block group level. We used spatial data and cross-classification analysis to compare the prevalence of industrial hog operations in neighborhoods that are primarily African American and low income with the prevalence in neighborhoods that are African American and affluent. We also used logistic regression to evaluate the relationship between the environmental justice variables and the location of the industrial hog operations. The block group characterization showed a high prevalence of hog operations in the four highest quintiles compared with the lowest quintile for percentage African American and percentage poverty. At increasing levels of percentage African Americans and percentage of persons in poverty, there are 2.4-3.6 times more operations compared with the referent group; additionally, scale adjustment to only the hog counties reduces this to 1.8-3.1 more operations compared with the referent group. The inequitable distribution of hog-confined agricultural feeding operations in these communities may have adverse environmental impacts associated with industrial hog production, such as increased health risks and quality of life degradation, as have occurred in other areas having similar facilities. PMID- 11929729 TI - Evaluating cumulative risk assessment for environmental justice: a community case study. AB - A key feature of cumulative risk assessment (CRA) is the ability to estimate differential health risks from environmental exposures within populations. Identifying populations at increased risk from environmental exposures is the first step toward mitigating such risks as required by the fair treatment mandate of environmental justice. CRA methods remain under development except for a limited application in pesticide regulations. The goals of this research were to advance CRA methods and to test their application in a community case study. We compared cumulative risk and health assessments for South and Southwest Philadelphia communities. The analysis found positive correlations between cumulative risk and mortality measurements for total mortality in Whites and non Whites when we conducted the risk assessment using a multi-end point toxicological database developed for this project. Cumulative risk scores correlated positively with cause-specific mortality in non-Whites. Statistically significant increases in total and respiratory mortality rates were associated with incremental increases in the hazard ratio cumulative risk scores, with ranges of 2-6% for total and 8-23% for respiratory. Regression analyses controlled for percent non-White population and per capita income, indicating that risk scores represent an environmental effect on health independent of race and income. This case study demonstrated the successful application of CRA at the community level. CRA adds a health dimension to pollutant concentrations to produce a more comprehensive understanding of environmental inequities that can inform decision making. CRA is a viable tool to identify high-risk areas and to guide surveillance, research, or interventions. PMID- 11929731 TI - Overlooked and underserved in Harlem: a population-based survey of adults with asthma. AB - The prevalence of asthma has increased over the past two decades; if this trend persists over the next two decades, the number of individuals with asthma in the United States will double by 2020, affecting 29 million Americans. Many of these individuals will be adults. Recent community-based participatory research in Harlem has focused on children with asthma, but little is known about the prevalence and burden of asthma among adults. We conducted a population-based probability sample of Central Harlem adults 18-65 years of age from 1992 to 1994. Asthma was one of three ambulatory care-sensitive conditions surveyed. We used an additional set of questions regarding asthma management and burden for those respondents who reported they had asthma. The prevalence of self-reported asthma was 14% in this population-based sample of Central Harlem adults. Respondents with asthma reported remarkably high rates of emergency department (ED) visits for asthma, but women were more likely than men to report two or more ED visits in the year prior to interview (38% vs. 18%). Women with asthma were also more likely than men with asthma to report activity restrictions because of asthma (61% vs. 26%). The burden of asthma among adults in Central Harlem is considerable. We urgently need comprehensive health approaches to address the high prevalence of health risks related to multiple chronic diseases, notably smoking and obesity. Key priorities are to determine which community education, prevention, and promotion programs are most effective and will best serve Harlem adults. PMID- 11929730 TI - Different slopes for different folks: socioeconomic and racial/ethnic disparities in asthma and hay fever among 173,859 U.S. men and women. AB - Although allergic diseases such as asthma and hay fever are a major cause of morbidity in industrialized countries, most studies have focused on patterns of prevalence among children and adolescents, with relatively few studies on variations in prevalence by race/ethnicity and socioeconomic position among adults. Our study examined racial/ethnic and socioeconomic patterns in the prevalence of asthma overall, asthma with hay fever, asthma without hay fever, and hay fever overall, in a population of 173,859 women and men in a large prepaid health plan in northern California. Using education as a measure of socioeconomic position, we found evidence of a positive gradient for asthma with hay fever with increasing level of education but an inverse gradient for asthma without hay fever. Hay fever was also strongly associated with education. Compared with their White counterparts, Black women and men were more likely to report asthma without hay fever, and Black women were less likely to have asthma with hay fever. Asian men were also more likely to report asthma with hay fever, and Asian women and men were much more likely to have hay fever. Racial/ethnic disparities in prevalence of allergic diseases were largely independent of education. We discuss implications for understanding these social inequalities in allergic disease risk in relation to possible differences in exposure to allergens and determinants of immunologic susceptibility and suggest directions for future research. PMID- 11929733 TI - Pesticide safety among farmworkers: perceived risk and perceived control as factors reflecting environmental justice. AB - Farmworkers in the United States constitute a population at risk for serious environmental and occupational illness and injury as well as health disparities typically associated with poverty. Pesticides are a major source of occupational injury and illness to which farmworkers are exposed. Efforts to provide safety training for farmworkers have not been fully evaluated. Based on the Health Belief Model, this analysis examines how safety information affects perceived pesticide safety risk and control among farmworkers and how perceived risk and control affect farmworker knowledge and safety behavior. Data are based on interviews conducted in 1999 with 293 farmworkers in eastern North Carolina as part of the Preventing Agricultural Chemical Exposure in North Carolina Farmworkers' Project. Perceived pesticide risk and perceived pesticide control scales were developed from interview items. Analysis of the items and scales showed that farmworkers had fairly high levels of perceived risk from pesticides and perceived control of pesticide safety. Receiving information about pesticide safety (e.g., warning signs) reduced perceived risk and increased perceived control. Pesticide exposure knowledge was strongly related to perceived risk. However, perceived risk had a limited relationship to safety knowledge and was not related to safety behavior. Perceived control was not related to pesticide exposure knowledge, but was strongly related to safety knowledge and safety behavior. A key tenet of environmental justice is that communities must have control over their environment. These results argue that for pesticide safety education to be effective, it must address issues of farmworker control in implementing workplace pesticide safety. PMID- 11929732 TI - Lead sources, behaviors, and socioeconomic factors in relation to blood lead of native american and white children: a community-based assessment of a former mining area. AB - Lead poisoning prevention requires knowledge of lead sources and of appropriate residential lead standards. Data are severely lacking on lead sources for Native American children, many of whom live in rural areas. Further, the relation of mining waste to blood lead concentrations (BPbs) of rural children is controversial. In collaboration with the eight tribes of northeastern Oklahoma, we assessed lead sources and their effects on BPbs for rural Native American and White children living in a former mining region. Venous blood lead, residential environmental (soil, dust, paint, water), and caregiver interview (e.g., hand-to mouth behaviors, socioeconomic conditions) data were obtained from a representative sample of 245 children 1-6 years of age. BPbs ranged from 1 to 24 microg/dL. There were no ethnic differences in BPbs (p= 0.48) nor any patterns of excess lead sources for Native American or White children. Multiple linear regression analyses indicated that mean soil lead, mean floor lead loading, mouthing behaviors, caregivers' education, and residence in former mining towns were all strongly associated with BPbs. Logistic regression results showed mean floor dust lead loading greater than or equal to 10.1 microg/ft(2) (odds ratio [OR], 11.4; 95% confidence interval [CI], 3.5-37.3), and yard soil lead >165.3 mg/kg (OR, 4.1; CI, 1.3-12.4) were independently associated with BPbs greater than or equal to 10 microg/dL. We also found strong interactions between soil lead and poverty (p= 0.005), and dust and soil sources (p= 0.02). Our findings indicate that soil and dust lead derived largely from mining waste pose a health hazard to Native American and White children, and that current residential dust lead standards are insufficient to adequately protect children. Moreover, our finding that poor children are especially vulnerable to lead exposures suggests that residential standards should consider interactions among socioeconomic conditions and lead sources if environmental justice is to be achieved. PMID- 11929734 TI - Combining community-based research and local knowledge to confront asthma and subsistence-fishing hazards in Greenpoint/Williamsburg, Brooklyn, New York. AB - Activists in the environmental justice movement are challenging expert-driven scientific research by taking the research process into their own hands and speaking for themselves by defining, analyzing, and prescribing solutions for the environmental health hazards confronting communities of the poor and people of color. I highlight the work of El Puente and The Watchperson Project--two community-based organizations in the Greenpoint/Williamsburg neighborhood in Brooklyn, New York, that have engaged in community-based participatory research (CBPR) to address asthma and risks from subsistence-fish diets. The CBPR process aims to engage community members as equal partners alongside scientists in problem definition, information collection, and data analysis--all geared toward locally relevant action for social change. In the first case I highlight how El Puente has organized residents to conduct a series of asthma health surveys and tapped into local knowledge of the Latino population to understand potential asthma triggers and to devise culturally relevant health interventions. In a second case I follow The Watchperson Project and their work surveying subsistence anglers and note how the community-gathered information contributed key data inputs for the U.S. Environmental Protection Agency Cumulative Exposure Project in the neighborhood. In each case I review the processes each organization used to conduct CBPR, some of their findings, and the local knowledge they gathered, all of which were crucial for understanding and addressing local environmental health issues. I conclude with some observations about the benefits and limits of CBPR for helping scientists and communities pursue environmental justice. PMID- 11929736 TI - Holistic risk-based environmental decision making: a Native perspective. AB - Native American Nations have become increasingly concerned about the impacts of toxic substances. Although risk assessment and risk management processes have been used by government agencies to help estimate and manage risks associated with exposure to toxicants, these tools have many inadequacies and as a result have not served Native people well. In addition, resources have not always been adequate to address the concerns of Native Nations, and involvement of Native decision makers on a government-to-government basis in discussions regarding risk has only recently become common. Finally, because the definitions of health used by Native people are strikingly different from that of risk assessors, there is also a need to expand current definitions and incorporate traditional knowledge into decision making. Examples are discussed from the First Environment Restoration Initiative, a project that is working to address toxicant issues facing the Mohawk territory of Akwesasne. This project is developing a community defined model in which health is protected at the same time that traditional cultural practices, which have long been the key to individual and community health, are maintained and restored. PMID- 11929735 TI - Is the U.S. Environmental Protection Agency brownfields assessment pilot program environmentally just? AB - In the early 1990s, the U.S. Environmental Protection Agency (U.S. EPA) started a grant program to assist communities redevelop brownfields, which are abandoned or underutilized sites that have real or perceived contamination. In addition to determining if the communities receiving the grants were the most distressed cities in the United States, we also evaluate the U.S. EPA program in terms of environmental justice at the macro scale. Using 1990 U.S. Census of Housing and Population data and a matched-cities methodology, we compared the brownfields pilot cities to other communities in the United States. We found that regardless of intent, the U.S. EPA program is environmentally just by disproportionately awarding grants to the most economically distressed cities. We also found that the cities that received funding in the early years of the program were more economically distressed than cities receiving the funding more recently. PMID- 11929737 TI - "Hey, mom, thanks!": use of focus groups in the development of place-specific materials for a community environmental action campaign. AB - We examined the relevance of five strategies to reduce the risk of exposure to environmental hazards for African-American and Hispanic children living in Northern Manhattan in New York City. Researchers conducting a community-wide intervention to increase awareness of environmental health hazards identified five strategies for keeping children healthy, preventing asthma, and promoting children's growth and development. These strategies were based on current scientific knowledge of environmental health and were tested and refined through a series of focus groups. The 14 focus groups were conducted with women of childbearing age living in the communities under study. The purpose of the focus groups was to test the relevancy of the five strategies and to obtain data to inform the intervention's social action campaign. Here authors discuss the process of identifying strategies for risk reduction and incorporating community residents' perceptions of risk into health risk messages. The authors argue that broader social and historical contexts are important in shaping community members' interpretations of risk and subsequent response to health education campaigns. PMID- 11929738 TI - Awareness of environmental risks and protective actions among minority women in Northern Manhattan. AB - We report findings of a survey of 555 women 18-35 years of age living in Northern Manhattan in New York City. The survey was conducted by the Columbia Center for Children's Environmental Health (CCCEH) to learn what community women knew about environmental risks to health and what they did to protect themselves and their families, to validate the findings of focus groups held with community women, and to provide information for planning the Healthy Home, Healthy Child campaign sponsored by CCCEH. Survey findings showed that overall awareness of environmental risks to children's health was high, with more than 95% of respondents identifying lead, household pests, pesticides, environmental tobacco smoke, and drugs as harmful to health. Similarly, more than 95% of respondents reported taking one or more protective actions to reduce these risks, suggesting that these factors significantly concern women living in Northern Manhattan. The reported levels of specific protective actions to reduce these risks, however, varied greatly. In each area of risk the most frequently reported actions were effective ones, but many other important protective actions were rarely mentioned, suggesting that there was room for an educational campaign to teach women new ways to protect their families. Survey respondents and CCCEH scientists differed in the priorities they placed on the importance of key protective actions, confirming earlier focus group findings and suggesting the importance of incorporating community concerns into the planning of environmental campaigns. PMID- 11929739 TI - Unequal exposure to ecological hazards: environmental injustices in the Commonwealth of Massachusetts. AB - This study analyzes the social and geographic distribution of ecological hazards across 368 communities in the Commonwealth of Massachusetts. Combining census data with a variety of environmental data, we tested for and identified both income-based and racially based biases to the geographic distribution of 17 different types of environmentally hazardous sites and industrial facilities. We also developed a composite measure of cumulative exposure to compare the relative overall risks characteristic of each community. To the best of our knowledge, this point system makes this the first environmental justice study to develop a means for measuring and ranking cumulative exposure for communities. The study also controls for the intensity of hazards in each community by accounting for the area across which hazards are distributed. The findings indicate that ecologically hazardous sites and facilities are disproportionately located and concentrated in communities of color and working-class communities. The implication of this research for policymakers and citizen advocates is that cumulative exposure of residents to environmentally hazardous facilities and sites should receive greater consideration regarding community demographics and environmental health indicators. We conclude that the provision of additional resources for environmental monitoring and ranking, as well as yearly progress reports, is necessary for communities and state agencies to achieve equal access to clean and healthy environments for all residents. PMID- 11929740 TI - Segregation and black/white differences in exposure to air toxics in 1990. AB - I examined non-Hispanic Black and non-Hispanic White differences in exposure to noncriteria air pollutants in 44 U.S. Census Bureau-defined metropolitan areas with populations greater than one million, using data on air toxics concentrations prepared for the U.S. Environmental Protection Agency as part of its Cumulative Exposure Project combined with U.S. census data. I measured differences in exposure to air toxics through the calculation of a net difference score, which is a statistical measure used in income inequality analysis to measure inequality over the whole range of exposures. The scores ranged from 11.52 to 83.60. In every metropolitan area, non-Hispanic Blacks are more likely than non-Hispanic Whites to be living in tracts with higher total modeled air toxics concentrations. To assess potential reasons for such a wide variation in exposure differences, I performed a multiple regression analysis with the net difference score as the dependent variable. Independent variables initially included were as follows: the dissimilarity index (to measure segregation), Black poverty/White poverty (to control for Black/White economic differences), population density and percentage of persons traveling to work who drive to work (alone and in car pools), and percentage of workforce employed in manufacturing (factors affecting air quality). After an initial analysis I eliminated from the model the measures of density and the persons driving to work because they were statistically insignificant, they did not add to the predictive power of the model, and their deletion did not affect the other variables. The final model had an R(2) of 0.56. Increased segregation is associated with increased disparity in potential exposure to air pollution. PMID- 11929741 TI - From asthma to AirBeat: community-driven monitoring of fine particles and black carbon in Roxbury, Massachusetts. AB - Asthma is an ongoing environmental justice concern in Roxbury, an urban neighborhood of Boston, Massachusetts. Residents, especially local youth, were the first to investigate the potential links between high asthma rates and air pollution, particularly from diesel buses and trucks. A youth-led march for clean air and community air monitoring projects drew governmental and media attention to these problems. In 1998, a collaboration of environmental justice, government, and research groups came together to develop a real-time air pollution monitoring system known as AirBeat. This community-based participatory research project was designed to answer community questions about whether there are pollution "hot spots" in Roxbury and the degree to which diesel emissions are contributing to health problems. AirBeat measures and reports levels of PM2.5 (particulate matter with a mass median aerodynamic diameter < or= to 2.5 microm), ozone, and black carbon on an hourly basis. These data are accessible via a website, telephone hotline, and a flag warning system. AirBeat is successful because community residents and organizations participate as equal partners with an equitable share of funding. The project also promotes a community sense of ownership and pride. Dozens of youth have developed leadership and scientific skills. The media have extensively covered the project as a community victory. The data support the claim that Dudley Square in Roxbury is a hot spot for air pollution. This information is now being used to advocate for alternative fuel transit buses and other clean air measures. Finally, this project has strengthened community partnerships with research and governmental institutions. PMID- 11929742 TI - Do Mexican Americans perceive environmental issues differently than Caucasians: a study of cross-ethnic variation in perceptions related to water in Tucson. AB - Little is known about the environmental perceptions of our nation's Mexican and Mexican American population, especially in the area of water quality. We examined these perceptions to determine the extent to which Caucasians and Mexican Americans living in the Tucson, Arizona, metropolitan area differ in their perceptions of water quality-related risk, inequity, trust, and participation in civic activities. Ethnic variations in perceptions toward inequity, trust, and public participation were observed even when socioeconomic variation between Caucasians and Mexican Americans was controlled. However, significant ethnic variations in perceptions of water quality-related risks were observed only when socioeconomic variation was not controlled. Implications of these findings to environmental justice efforts in Mexican American communities are discussed. PMID- 11929744 TI - Deteriorated housing contributes to high cockroach allergen levels in inner-city households. AB - The high prevalence of childhood asthma in low-income, inner-city populations is not fully understood but has been at least partly attributed to the disproportionate exposures associated with socioeconomic disadvantage. The contribution of indoor allergens to asthma is well documented, but links between socioeconomic disadvantage and indoor allergen levels are not clear. We investigated levels of cockroach allergens (Bla g 2) in a sample of 132 Dominican or African American low-income households with young children in northern Manhattan in New York City (40% were receiving public assistance) to determine whether the distribution of allergens is a function of housing deterioration. Deterioration was measured by the presence and number of physical housing problems (holes in the ceilings and walls, water damage, etc.). More than 50% of the sample had two or more types of housing dilapidation, and 67% of the sample reported cockroach sightings in their homes. Samples of dust were collected from kitchen and bedroom surfaces. We hypothesized that the greater the dilapidation, the higher the allergen levels, independent of income, sociocultural factors, and pest-control methods. In addition, we hypothesized that the homes of families characterized by frequent moves (23.5%) would have higher allergen levels than more stable families. Results showed significant positive associations between housing deterioration and allergen levels in kitchens, after adjusting for income and ethnicity, with independent effects of residential stability (p< 0.05). Bedroom allergen levels were associated with housing instability (p < 0.01) and ethnicity (p< 0.01). Findings demonstrated that indoor household allergen levels are related to degree of household disrepair, after adjusting for individual family attributes, suggesting that social-structural aspects of housing may be appropriate targets for public health interventions designed to reduce allergen exposure. PMID- 11929743 TI - The Seattle-King County healthy homes project: implementation of a comprehensive approach to improving indoor environmental quality for low-income children with asthma. AB - Pediatric asthma is a growing public health issue, disproportionately affecting low-income people and people of color. Exposure to indoor asthma triggers plays an important role in the development and exacerbation of asthma. We describe the implementation of the Seattle-King County Healthy Homes Project, a randomized, controlled trial of an outreach/education intervention to improve asthma-related health status by reducing exposure to allergens and irritants in the home. We randomly assigned 274 low-income children with asthma ages 4-12 to either a high- or a low-intensity group. In the high-intensity group, community health workers called Community Home Environmental Specialists (CHES) conducted initial home environmental assessments, provided individualized action plans, and made additional visits over a 12-month period to provide education and social support, encouragement of participant actions, provision of materials to reduce exposures (including bedding encasements), assistance with roach and rodent eradication, and advocacy for improved housing conditions. Members of the low-intensity group received the initial assessment, home action plan, limited education during the assessment visit, and bedding encasements. We describe the recruitment and training of CHES and challenges they faced and explain the assessment and exposure reduction protocols addressing dust mites, mold, tobacco smoke, pets, cockroaches, rodents, dust, moisture, and toxic or hazardous chemicals. We also discuss the gap between the practices recommended in the literature and what is feasible in the home. We accomplished home interventions and participants found the project very useful. The project was limited in resolving structural housing quality issues that contributed to exposure to indoor triggers. PMID- 11929748 TI - Interactions of STAT5b-RARalpha, a novel acute promyelocytic leukemia fusion protein, with retinoic acid receptor and STAT3 signaling pathways. AB - Signal transducer and activator of transcription (STAT) 5b-retinoic acid receptor (RAR) alpha is the fifth fusion protein identified in acute promyelocytic leukemia (APL). Initially described in a patient with all-trans retinoic acid (ATRA)-unresponsive disease, STAT5b-RARalpha resulted from an interstitial deletion on chromosome 17. To determine the molecular mechanisms of myeloid leukemogenesis and maturation arrest in STAT5b-RARalpha(+) APL and its unresponsiveness to ATRA, we examined the effect of STAT5b-RARalpha on the activity of myeloid transcription factors including RARalpha/retinoid X receptor (RXR) alpha, STAT3, and STAT5 as well as its molecular interactions with the nuclear receptor corepressor, SMRT, and nuclear receptor coactivator, TRAM-1. STAT5b-RARalpha bound to retinoic acid response elements (RAREs) both as a homodimer and as a heterodimer with RXRalpha and inhibited wild-type RARalpha/RXRalpha transactivation. Although STAT5b-RARalpha had no effect on ligand-induced STAT5b activation, it enhanced interleukin 6-induced STAT3 dependent reporter activity, an effect shared by other APL fusion proteins including promyelocytic leukemia-RARalpha and promyelocytic leukemia zinc finger (PLZF)-RARalpha. SMRT was released from STAT5b-RARalpha/SMRT complexes by ATRA at 10(-6) M, whereas TRAM-1 became associated with STAT5b-RARalpha at 10(-7) M. The coiled-coil domain of STAT5b was required for formation of STAT5b-RARalpha homodimers, for the inhibition of RARalpha/RXRalpha transcriptional activity, and for stability of the STAT5b-RARalpha/SMRT complex. Thus, STAT5b-RARalpha contributes to myeloid maturation arrest by binding to RARE as either a homodimer or as a heterodimer with RXRalpha resulting in the recruitment of SMRT and inhibition of RARalpha/RXRalpha transcriptional activity. In addition, STAT5b RARalpha and other APL fusion proteins may contribute to leukemogenesis by interaction with the STAT3 oncogene pathway. PMID- 11929749 TI - The Stat5-RARalpha fusion protein represses transcription and differentiation through interaction with a corepressor complex. AB - The transcription factor Stat5 mediates the cellular response to activation of multiple cytokine receptors involved in the regulation of proliferation and differentiation of hematopoietic cells. Recently, the human Stat5 gene was found to be translocated to the RARalpha gene in a patient with acute promyelocytic leukemia indicating that Stat5 might also play a role in cellular transformation. We investigated the mechanism by which Stat5 might exert this function and studied the biochemical and cellular functions of fusion proteins comprising Stat5 and RARalpha. The expression of Stat5-RARalpha causes the transcriptional repression of gene transcription, a process that requires the coiled-coil domain of Stat5 (amino acid positions 133-333). Oligomerization of this domain in the Stat5-RARalpha fusion protein leads to stable binding of the corepressor SMRT independent of all-trans retinoic acid (ATRA) stimulation and is accompanied by an impaired response to differentiation signals in hematopoietic cells. This inhibitory effect on myeloid differentiation cannot be overcome by simultaneous coexpression of RARalpha. We conclude that Stat5 is capable of interacting with a corepressor complex that alters the pattern of corepressor binding to RARalpha and its dissociation in response to ATRA stimulation, leading to enhanced repressor activity and a block of hematopoietic differentiation. PMID- 11929750 TI - Creation of a genetic system for analysis of the phagocyte respiratory burst: high-level reconstitution of the NADPH oxidase in a nonhematopoietic system. AB - The phagocyte nicotinamide adenine dinucleotide phosphate (reduced form) (NADPH) oxidase was functionally reconstituted in monkey kidney COS-7 cells by transfection of essential subunits, gp91(phox), p22(phox), p47(phox), and p67(phox). COS-7 cells express the essential small guanosine 5'-triphosphatase, Rac1. Transgenic COS-phox cells were capable of arachidonic acid-induced NADPH oxidase activity up to 80% of that of human neutrophils, and of phorbol myristate acetate (PMA)-induced activity up to 20% of that of neutrophils. Expression of all 4 phox components was required for enzyme activity, and enzyme activation was associated with membrane translocation of p47(phox), p67(phox), and Rac1. Expression of p47(phox) Ser303Ala/Ser304Ala or Ser379Ala phosphorylation deficient mutants resulted in significantly impaired NAPDH oxidase activity, compared with expression of wild-type p47(phox) or the p47(phox) Ser303Glu/Ser304Glu phosphorylation mimic, suggesting that p47(phox) phosphorylation contributes to enzyme activity in the COS system, as is the case in neutrophils. Hence, COS-phox cells should be useful as a new whole-cell model that is both capable of high-level superoxide production and readily amenable to genetic manipulation for investigation of NADPH oxidase function. PMA-elicited superoxide production in COS-phox cells was regulated by activation of protein kinase C (PKC) and Rac. Although COS-7 cells differ from human neutrophils in PKC isoform expression, transient expression of major neutrophil isoforms in COS-phox cells did not increase PMA-induced superoxide production, suggesting that endogenous isoforms were not rate limiting. Val204 in p67(phox), previously shown to be required for NADPH oxidase activity under cell-free conditions, was found to be essential for superoxide production by intact COS-phox cells, on the basis of transfection studies using a p67(phox) (Val204Ala) mutant. PMID- 11929751 TI - IRTAs: a new family of immunoglobulinlike receptors differentially expressed in B cells. AB - The IRTA1 and IRTA2 genes encode immunoglobulinlike cell surface receptors expressed in B cells and involved in chromosome 1q21 translocations in B-cell malignancy. We have now characterized and comparatively analyzed the structure and expression pattern of the entire family of IRTA genes, which includes 5 members contiguously located on chromosome 1q21. The IRTA messenger RNAs are expressed predominantly in the B-cell lineage within discrete B-cell compartments: IRTA1 is specific to the marginal zone, IRTA2 and IRTA3 are found in the germinal center light zone and in intraepithelial and interfollicular regions, and IRTA4 and IRTA5 are expressed predominantly in the mantle zone. All IRTA genes code for transmembrane receptors that are closely related to Fc receptors in their most amino-terminal extracellular domains and that possess cytoplasmic domains containing ITIM (immunotyrosine inhibition motifs)- and, possibly, ITAM (immunotyrosine activation motifs)-like motifs. These structural features suggest that the IRTA receptors may play a role in regulating activation of normal B cells and possibly in the development of neoplasia. PMID- 11929752 TI - Sustained phenotypic correction of hemophilia B dogs with a factor IX null mutation by liver-directed gene therapy. AB - Hemophilia B is an X-linked coagulopathy caused by absence of functional coagulation factor IX (FIX). Using adeno-associated virus (AAV)-mediated, liver directed gene therapy, we achieved long-term (> 17 months) substantial correction of canine hemophilia B in 3 of 4 animals, including 2 dogs with an FIX null mutation. This was accomplished with a comparatively low dose of 1 x 10(12) vector genomes/kg. Canine FIX (cFIX) levels rose to 5% to 12% of normal, high enough to result in nearly complete phenotypic correction of the disease. Activated clotting times and whole blood clotting times were normalized, activated partial thromboplastin times were substantially reduced, and anti-cFIX was not detected. The fourth animal, also a null mutation dog, showed transient expression (4 weeks), but subsequently developed neutralizing anti-cFIX (inhibitor). Previous work in the canine null mutation model has invariably resulted in inhibitor formation following treatment by either gene or protein replacement therapies. This study demonstrates that hepatic AAV gene transfer can result in sustained therapeutic expression in a large animal model characterized by increased risk of a neutralizing anti-FIX response. PMID- 11929753 TI - Plasmodium yoelii uses the murine Duffy antigen receptor for chemokines as a receptor for normocyte invasion and an alternative receptor for reticulocyte invasion. AB - Erythrocyte invasion by malaria parasites is a complex multistep process involving parasite and erythrocyte receptors. It is a critical stage in the parasite life cycle and, therefore, a logical step in which to intervene to prevent or ameliorate disease. Rodent models of malaria, commonly Plasmodium yoelii, are frequently used for studies of malaria pathogenesis. Little is known, however, about the invasion machinery of rodent malaria parasites. We have found previously that mice congenic for a region of chromosome 1, containing the Duffy antigen/receptor for chemokines (DARC), have different susceptibility to P yoelii infection. Because P vivax, a human parasite, and P knowlesi, a simian parasite, use DARC to enter human erythrocytes, we sought to identify the role of the murine DARC in P yoelii invasion. Using a novel in vivo invasion assay and DARC knock-out mice, we found that DARC knock-out normocytes (mature erythrocytes) had negligible levels of P yoelii invasion compared with wild-type normocytes, demonstrating that DARC is a receptor for invasion of murine erythrocytes. In contrast, DARC knock-out reticulocytes were invaded at a rate similar to that for wild-type reticulocytes. We conclude that there is a DARC- independent pathway for reticulocyte invasion. These findings represent the first identification of a murine malaria receptor on erythrocytes and the first determination that different pathways of invasion exist on normocytes and reticulocytes. Because we show conservation of host-receptor interactions between rodent and human malaria, we can now use this model to identify how immunity can interfere with the invasion process. PMID- 11929755 TI - Autoimmunity in human primary immunodeficiency diseases. AB - Human primary immunodeficiency diseases are experiments of nature characterized by an increased susceptibility to infection. In many cases, they are also associated with troublesome and sometimes life-threatening autoimmune complications. In the past few years, great strides have been made in understanding the molecular basis of primary immunodeficiencies, and this had led to more focused and successful treatment. This review has 3 aims: (1) to highlight the variety of autoimmune phenomena associated with human primary immunodeficiency diseases; (2) to explore how primary immunodeficiencies predispose patients to autoimmune phenomena triggered by opportunistic infections; and (3) to consider the rationale for the current treatment strategies for autoimmune phenomena, specifically in relation to primary immunodeficiency diseases. Reviewing recent advances in our understanding of the small subgroup of patients with defined causes for their autoimmunity may lead to the development of more effective treatment strategies for idiopathic human autoimmune diseases. PMID- 11929754 TI - Dose-adjusted EPOCH chemotherapy for untreated large B-cell lymphomas: a pharmacodynamic approach with high efficacy. AB - We hypothesized that incremental improvements in the cyclophosphamide-doxorubicin vincristine-prednisone (CHOP) chemotherapy regimen through optimization of drug selection, schedule, and pharmacokinetics would improve outcome in patients with large B-cell lymphomas. A prospective multi-institutional study of administration of etoposide, vincristine, and doxorubicin for 96 hours with bolus doses of cyclophosphamide and oral prednisone (EPOCH therapy) was done in 50 patients with previously untreated large B-cell lymphomas. The doses of etoposide, doxorubicin, and cyclophosphamide were adjusted 20% each cycle to achieve a nadir absolute neutrophil count below 0.5 x 10(9)/L. The median age of the patients was 46 years (range, 20-88 years); 24% were older than 60 years; and 44% were at high intermediate or high risk according to International Prognostic Index (IPI) criteria. There was a complete response in 92% of patients, and at the median follow-up time of 62 months, the progression-free survival (PFS) and overall survival (OS) rates were 70% and 73%, respectively. Neither IPI risk factors nor the index itself was associated with response, PFS, or OS. Doses were escalated in 58% of cycles, and toxicity levels were tolerable. Significant inverse correlations were observed between dose intensity and age for all adjusted agents, and drug clearance of doxorubicin and free etoposide was also inversely correlated with age (r = -0.54 and P(2) =.08 and r = -0.45 and P(2) =.034, respectively). Free-etoposide clearance increased significantly during successive cycles (P(2) =.015). Lymphomas with proliferation of at least 80% had somewhat lower progression and those expressing bcl-2 had significantly higher progression (P(2) =.04). Expression of bcl-2 may discriminate the recently described activated B-like from germinal-center B-like large-cell lymphomas and provide important pathobiologic and prognostic information. Dose-adjusted EPOCH may produce more cell kill than CHOP-based regimens. Dynamic dose adjustment may overcome inadequate drug concentrations, particularly in younger patients, and compensate for increased drug clearance over time. PMID- 11929756 TI - Regulation of endothelial cell branching morphogenesis by endogenous chemokine stromal-derived factor-1. AB - The chemokine stromal-derived factor-1 (SDF-1) and its unique receptor, CXCR4, are required for normal cardiovascular development, but a critical role for SDF-1 in postnatal vascular remodeling and the mechanisms underlying SDF-1/CXCR-4 vasculogenesis are unclear. Here we show that SDF-1 is expressed by the vascular endothelium from selected healthy and tumor tissues. In vitro, primary endothelial cells constitutively express SDF-1 that is detected in the cytoplasm, on the cell surface, and in the culture supernatant. Vascular endothelial growth factor (VEGF) and basic fibroblast growth factor (bFGF) increase SDF-1 expression in endothelial cells. In functional studies, pertussis toxin and antibodies to SDF-1 or CXCR-4 disrupt extracellular matrix-dependent endothelial cell tube formation in vitro. This morphogenic process is associated with time-dependent modulation of surface CXCR-4 expression that changes from being diffuse to being polarized and subsequently lost. In vivo, pertussis toxin and neutralizing antibodies directed at SDF-1 inhibit growth factor-dependent neovascularization. These results indicate that SDF-1/CXCR-4 identifies VEGF- and bFGF-regulated autocrine signaling systems that are essential regulators of endothelial cell morphogenesis and angiogenesis. PMID- 11929757 TI - A humanized non-FcR-binding anti-CD3 antibody, visilizumab, for treatment of steroid-refractory acute graft-versus-host disease. AB - Visilizumab is a humanized anti-CD3 monoclonal antibody characterized by a mutated IgG2 isotype, lack of binding to Fcgamma-receptors, and ability to induce apoptosis selectively in activated T cells. To test pharmacokinetics, safety, and immunosuppressive activity of visilizumab, 17 patients with glucocorticoid refractory acute graft-versus-host disease (GVHD) were enrolled in a phase 1 study. Six patients were given 7 doses of visilizumab (0.25 or 1.0 mg/m(2)) on days 1, 3, 5, 7, 9, 11, and 13. Because multiple doses of 1 mg/m(2) caused delayed visilizumab accumulation and prolonged lymphopenia, the next 11 patients received a single dose of 3.0 mg/m(2) on day 1. GVHD improved in all patients; 15 were evaluable through day 42. Multiple dosing resulted in 1 of 6 complete responses (CRs) and 5 partial responses (PRs), but all 6 patients died at a median of 87 days after starting visilizumab therapy. Single dosing resulted in 6 of 9 CRs, 3 PRs, and 7 of 11 patients surviving after 260 to 490 days (median, 359 days; P =.03). There were no allergic reactions and 3 grade 1 acute infusional toxicities. Plasma Epstein-Barr virus (EBV) DNA titers more than 1000 copies/mL and posttransplant lymphoproliferative disease (PTLD) developed in 2 of the first 7 patients. Based on rising EBV DNA titers, 5 of the next 10 patients were given the B cell-specific monoclonal antibody, rituximab. EBV DNA became undetectable and no overt PTLD developed. Visilizumab is well tolerated and has activity in advanced GVHD. A phase 2 study incorporating preemptive therapy for PTLD is warranted to determine the efficacy of visilizumab in GVHD. PMID- 11929758 TI - A prospective study of venous thromboembolism in relation to factor V Leiden and related factors. AB - The aim of this study was to examine the occurrence of venous thromboembolism (VTE) in relation to factor V-related risk factors. Using a nested case-control design combining 2 population-based prospective studies, we measured factor V Leiden, HR2 haplotype, activated protein C (APC) resistance, and plasma factor V antigen in 335 participants who developed VTE during 8 years of follow-up and 688 controls. The overall odds ratio (OR) of VTE was 3.67 (95% CI, 2.20-6.12) in participants carrying factor V Leiden compared with noncarriers. APC resistance measured after predilution with factor V-deficient plasma conferred an OR of 2.58 (95% CI, 1.62-4.10). All 3 participants homozygous for the HR2 haplotype had a VTE, and the OR of VTE for homozygosity was estimated to be 5.5 (95% CI, 2.45 12.5). Carriers of the HR2 haplotype otherwise were not at increased risk of VTE overall (OR = 1.05; 95% CI, 0.64-1.72), but double heterozygotes for HR2 and factor V Leiden carried an OR of idiopathic VTE of 16.3 (95% CI, 1.7-159) compared with noncarriers. Factor V antigen also was not associated with VTE overall, but for participants with the combination of high factor V antigen plus factor V Leiden the OR of idiopathic VTE was 11.5 (95% CI, 4.2-31.4). In the general population, APC resistance and factor V Leiden were important VTE risk factors; homozygosity for the HR2 haplotype may be a risk factor but was rare; otherwise, HR2 haplotype and factor V antigen were not risk factors except in carriers of factor V Leiden. PMID- 11929759 TI - Association of CD34 cell dose with hematopoietic recovery, infections, and other outcomes after HLA-identical sibling bone marrow transplantation. AB - Although CD34 cell dose is known to influence outcome of peripheral stem cell- and/or T-cell-depleted transplantation, such data on unmanipulated marrow transplantation are scarce. To study the influence of CD34(+) cell dose on hematopoietic reconstitution and incidence of infections after bone marrow transplantation, we retrospectively analyzed 212 patients from January 1994 to August 1999 who received a transplant of an unmanipulated graft from an HLA identical sibling donor. Median age was 31 years; 176 patients had hematologic malignancies. Acute graft-versus-host disease prophylaxis consisted mainly in cyclosporin associated with methotrexate (n = 174). Median number of bone marrow nucleated cells and CD34(+) cells infused were 2.4 x 10(8)/kg and 3.7 x 10(6)/kg, respectively. A CD34(+) cell dose of 3 x 10(6)/kg or more significantly influenced neutrophil (hazard ratio [HR] = 1.37, P =.04), monocyte (HR = 1.47, P =.02), lymphocyte (HR = 1.70, P =.003), erythrocyte (HR = 1.77, P =.0002), and platelet (HR = 1.98, P =.00008) recoveries. CD34(+) cell dose also influenced the incidence of secondary neutropenia (HR = 0.60, P =.05). Bacterial and viral infections were not influenced by CD34 cell dose, whereas it influenced the incidence of fungal infections (HR = 0.41, P =.008). Estimated 180-day transplantation-related mortality (TRM) and 5-year survival were 25% and 56%, respectively, and both were highly affected by CD34(+) cell dose (HR = 0.55, P =.006 and HR = 0.54, P =.03, respectively). Five-year survival and 180-day TRM were, respectively, 64% and 19% for patients receiving a CD34(+) cell dose of 3 x 10(6)/kg or more and 40% and 37% for the remainders. In conclusion a CD34(+) cell dose of 3 x 10(6)/kg or more improved all hematopoietic recoveries, decreased the incidence of fungal infections and TRM, and improved overall survival. PMID- 11929760 TI - Comparison of Escherichia coli-asparaginase with Erwinia-asparaginase in the treatment of childhood lymphoid malignancies: results of a randomized European Organisation for Research and Treatment of Cancer-Children's Leukemia Group phase 3 trial. AB - Asparaginase is an enzyme used in the treatment of acute lymphoblastic leukemia and lymphoblastic lymphoma in children. It has minimal bone marrow toxicity. Its major side effects are anaphylaxis, pancreatitis, diabetes, coagulation abnormalities, and thrombosis, especially intracranial. It is derived from 2 different sources: Escherichia coli and Erwinia chrysanthemi. Nonrandomized clinical studies have suggested a similar efficacy of these 2 types of asparaginases and a lower toxicity for Erwinia-asparaginase. The European Organisation for Research and Treatment of Cancer-Children's Leukemia Group (EORTC-CLG) 58881 trial randomized 700 children with acute lymphoblastic leukemia or lymphoblastic lymphoma to either E coli- or Erwinia-asparaginase at the same dosage of 10 000 IU/m(2) twice weekly to compare toxicity and efficacy. Coagulation abnormalities were more frequent in the E coli-asparaginase than in the Erwinia-asparaginase arm of the study (30.2% versus 11.9%, P <.0001). The incidence of other toxicity was not significantly different. In the Erwinia asparaginase arm, more patients failed to achieve complete remission (4.9% versus 2.0%; P =.038) and the relapse rate was higher, leading to shorter event-free survival (hazard ratio,1.59; 95% CI, 1.23-2.06; P =.0004). The estimate of event free survival rate (SE) at 6 years was 59.8% (2.6%) versus 73.4% (2.4%). Overall survival rate at 6 years was also lower in the Erwinia-asparaginase arm at 75.1% (2.3%) versus 83.9% (2.0%), P =.002. With the dose scheduling used in this protocol, E coli-asparaginase induced more coagulation abnormalities but was superior to Erwinia-asparaginase for the treatment of childhood lymphoid malignancies. PMID- 11929761 TI - Receptor tyrosine kinase, EphB4 (HTK), accelerates differentiation of select human hematopoietic cells. AB - EphB4 (HTK) and its ligand, ephrinB2, are critical for angiogenesis and result in fatal abnormalities of capillary formation in null mice. EphB4 was originally identified in human bone marrow CD34(+) cells by us and has since been reported to be expressed in erythroid progenitors, whereas the ligand ephrinB2 is expressed in bone marrow stromal cells. Reasoning that the developmental relationship between angiogenesis and hematopoiesis implies common regulatory molecules, we assessed whether EphB4 signaling influences the function and phenotype of primitive human hematopoietic cells. Ectopically expressed EphB4 in cell lines of restricted differentiation potential promoted megakaryocytic differentiation, but not granulocytic or monocytic differentiation. Primary cord blood CD34(+) cells transduced with EphB4 resulted in the elevated expression of megakaryocytic and erythroid specific markers, consistent with EphB4 selectively enhancing some lineage-committed progenitors. In less mature cells, EphB4 depleted primitive cells, as measured by long-term culture-initiating cells or CD34(+)CD38(-) cell numbers, and increased progenitor cells of multiple cell types. Effects of ectopic EphB4 expression could be abrogated by either targeted mutations of select tyrosine residues or by the tyrosine kinase inhibitor, genistein. These data indicate that EphB4 accelerates the differentiation of primitive cells in a nonlineage-restricted manner but alters only select progenitor populations, influencing lineages linked by common ancestry with endothelial cells. EphB4 enforces preferential megakaryocytic and erythroid differentiation and may be a molecular bridge between angiogenesis and hematopoiesis. PMID- 11929762 TI - Long-term support of hematopoiesis by a single stem cell clone in patients with paroxysmal nocturnal hemoglobinuria. AB - Paroxysmal nocturnal hemoglobinuria (PNH) is an acquired hematopoietic stem cell disorder characterized by clonal blood cells that are deficient in glycosylphosphatidylinositol-anchored proteins because of somatic mutations of the PIG-A gene. Many patients with PNH have more than one PNH clone, but it is unclear whether a single PNH clone remains dominant or minor clones eventually become dominant. Furthermore, it is unknown how many hematopoietic stem cells (HSCs) sustain hematopoiesis and how long a single HSC can support hematopoiesis in humans. To understand dynamics of HSCs, we reanalyzed the PIG-A gene mutations in 9 patients 6 to 10 years after the previous analyses. The proportion of affected peripheral blood polymorphonuclear cells (PMNs) in each patient was highly variable; it increased in 2 (from 50% and 65% to 98% and 97%, respectively), was stable in 4 (changed less than 20%), and diminished in 3 (94%, 99%, and 98% to 33%, 57%, and 43%, respectively) patients. The complexity of these results reflects the high variability of the clinical course of PNH. In all patients, the previously predominant clone was still present and dominant. Therefore, one stem cell clone can sustain hematopoiesis for 6 to 10 years in patients with PNH. Two patients whose affected PMNs decreased because of a decline of the predominant PNH clone and who have been followed up for 24 and 31 years now have an aplastic condition, suggesting that aplasia is a terminal feature of PNH. PMID- 11929763 TI - Intrathymic and extrathymic development of human plasmacytoid dendritic cell precursors in vivo. AB - The development of plasmacytoid dendritic cells (pDC2) from human CD34(+) stem cells in vivo was studied in RAG-2(-/-) interleukin (IL)-2Rgamma(-/-) mice that lack functional T and B cells and natural killer cells. CD34(+) cells isolated from fetal liver or thymus were labeled with 5- and 6-carboxyfluorescein diacetate succinimidyl ester (CFSE) and were injected into a human thymus grafted subcutaneously in the RAG-2(-/-) IL-2Rgamma(-/-) mice. One to 4 weeks later the CFSE label was found not only in T cells but also in CD123(+/high) CD4(+)CD45RA(+) pDC2, indicating that the CD34(+) cells can develop into pDC2 within a thymus. In addition to pDC2, CFSE-labeled dendritic cells with a mature phenotype, determined by the cell surface markers CD11c, CD83, and CD80, were found in the injected human thymus graft. pDC2 was not found in the periphery of mice carrying a human thymic graft, indicating that the intrathymic pDC2 failed to emigrate from the thymus. We also demonstrate that pDC2 can develop outside the thymus because relatively high percentages of pDC2 were found in the periphery after the intravenous injection of CD34(+)CD38(-) fetal liver cells in RAG-2(-/-) IL-2Rgamma(-/-) mice without a human thymus graft. These data indicate that the thymus and the peripheral pDC2 develop independently of each other. PMID- 11929764 TI - Extensive in vivo self-renewal, long-term reconstitution capacity, and hematopoietic multipotency of Pax5-deficient precursor B-cell clones. AB - Self-renewal, pluripotency, and long-term reconstitution are defining characteristics of single hematopoietic stem cells. Pax5(-/-) precursor B cells apparently possess similar characteristics. Here, using serial transplantations, with in vitro recloning and growth of the bone marrow-homed donor cells occurring after all transplantations, we analyzed the extent of self-renewal and hematopoietic multipotency of Pax5(-/-) precursor B-cell clones. Moreover, telomere length and telomerase activity in these clones was analyzed at various time points. Thus far, 5 successive transplantations have been performed. Clones transplanted for the fifth time, which have proliferated for more than 150 cell divisions in vitro, still repopulate the bone marrow with precursor B cells and reconstitute these recipients with lymphoid and myeloid cells. During this extensive proliferation, Pax5(-/-) precursor B cells shorten their telomeres at 70 to 90 base pairs per division. Their telomerase activity remains at 3% of that of HEK293 cancer cells during all serial in vivo transplantations/in vitro expansions. Together, these data show that Pax5(-/-) precursor B-cell clones possess extensive in vivo self-renewal capacity, long-term reconstitution capacity, and hematopoietic multipotency, with their telomeres shortening at the normal rate. PMID- 11929765 TI - Long-term repopulating ability of telomerase-deficient murine hematopoietic stem cells. AB - Telomere length must be tightly regulated in highly proliferative tissues, such as the lymphohematopoietic system. Under steady-state conditions, the levels and functionality of hematopoietic-committed or multipotent progenitors were not affected in late-generation telomerase-deficient mice (mTerc(-/-)) with critically short telomeres. Evaluation of self-renewal potential of mTerc(-/-) day-12 spleen colony-forming units demonstrated no alteration as compared with wildtype progenitors. However, the replating ability of mTerc(-/-) granulocyte macrophage CFUs (CFU-GMs) was greatly reduced as compared with wildtype CFU-GMs, indicating a diminished capacity of late-generation mTerc(-/-) committed progenitors when forced to proliferate. Long-term bone marrow cultures of mTerc( /-) bone marrow (BM) cells show a reduction in proliferative capacity; this defect can be mainly attributed to the hematopoietic, not to the stromal, mTerc( /-) cells. In serial and competitive transplantations, mTerc(-/-) BM stem cells show reduced long-term repopulating capacity, concomitant with an increase in genetic instability compared with wildtype cells. Nevertheless, in competitive transplantations late-generation mTerc(-/-) precursors can occasionally overcome this proliferative impairment and reconstitute irradiated recipients. In summary, our results demonstrate that late-generation mTerc(-/-) BM cells with short telomeres, although exhibiting reduced proliferation ability and reduced long term repopulating capacity, can still reconstitute myeloablated animals maintaining stem cell function. PMID- 11929766 TI - CCAAT/enhancer-binding proteins are required for granulopoiesis independent of their induction of the granulocyte colony-stimulating factor receptor. AB - Potential redundancy among members of the CCAAT/enhancer-binding protein (C/EBP) family in myeloid cells is indicated by the ability of C/EBPbeta to replace C/EBPalpha in vivo, by the expression of granulocyte colony-stimulating factor receptor (G-CSFR) on C/EBPalpha(-/-) cell lines, and by our finding that as with C/EBPalpha-estrogen receptor (C/EBPalpha-ER), either C/EBPbeta-ER or C/EBPdelta ER can induce terminal granulopoiesis in 32D cl3 cells. To assess the consequences of globally inhibiting C/EBPs, we employed KalphaER, containing a Kruppel-associated box (KRAB) transrepression domain, the C/EBPalpha DNA-binding domain, and an ER ligand-binding domain. C/EBPs have a common DNA-binding consensus, and activation of KalphaER repressed transactivation by endogenous C/EBPs 50-fold and reduced endogenous G-CSFR expression. In 32D cl3 cells coexpressing exogenous G-CSFR, activation of KalphaER prevented and even reversed myeloperoxidase, lysozyme, lactoferrin, and C/EBPepsilon RNA induction by G-CSF. In contrast, induction of PU.1 and CD11b, a gene regulated by PU.1 but not by C/EBPs, was unaffected. A KalphaER variant incapable of binding DNA owing to an altered leucine zipper did not affect 32D cl3 differentiation. Transduction of KalphaER into murine hematopoietic progenitor cells suppressed the formation of granulocyte colony-forming units, even in cytokines that enable C/EBPalpha(-/-) progenitors to differentiate into neutrophils. The formation of macrophage and of granulocyte-macrophage colony-forming units were also inhibited, but erythroid burst-forming units grew normally. Thus, in 32D cl3 cells and perhaps normal progenitors, C/EBPs are required for granulopoiesis beyond their ability to induce receptors for G-CSF and other cytokines. One requisite activity may be activation of the C/EBPepsilon gene by C/EBPalpha, as either C/EBPalpha-ER or C/EBPbeta-ER rapidly elevated C/EBPepsilon RNA in 32D cl3 cells in the presence of cycloheximide but not actinomycin D. PMID- 11929767 TI - Hematopoietic cells expressing the peripheral cannabinoid receptor migrate in response to the endocannabinoid 2-arachidonoylglycerol. AB - Cb2 is a novel protooncogene encoding the peripheral cannabinoid receptor. Previous studies demonstrated that 2 distinct noncoding first exons exist: exon 1A and exon-1B, which both splice to protein-coding exon-2. We demonstrate that in retrovirally induced murine myeloid leukemia cells with proviral insertion in Cb2, exon-1B/exon-2 Cb2 messenger RNA levels have been increased, resulting in high receptor numbers. In myeloid leukemia cells without virus insertion in this locus, low levels of only exon-1A/exon-2 Cb2 transcripts were present and receptors could not be detected. To elucidate the function of Cb2 in myeloid leukemia cells, a set of in vitro experiments was carried out using 32D/G-CSF-R (granulocyte colony-stimulating factor receptor) cells transfected with exon 1B/exon-2 Cb2 complementary DNA and a myeloid cell line carrying a virus insertion in Cb2 (ie, NFS 78). We demonstrate that a major function of the Cb2 receptor is stimulation of migration as determined in a transwell assay. Exposure of Cb2-expressing cells to different cannabinoids showed that the true ligand for Cb2 is 2-arachidonoylglycerol (2-AG), which may act as chemoattractant and as a chemokinetic agent. Furthermore, we observed a significant synergistic activity between 2-AG and interleukin-3 or G-CSF, suggesting cross-talk between the different receptor systems. Radioactive-ligand binding studies revealed significant numbers of Cb2 receptors in normal spleen. Transwell experiments carried out with normal mouse spleen cells showed 2-AG-induced migration of B220 , CD19-, immunoglobulin M-, and immunoglobulin D-expressing B lymphocytes. Our study demonstrates that a major function of Cb2 receptor expressed on myeloid leukemia cells or normal splenocytes is stimulation of migration. PMID- 11929768 TI - Coronary no-reflow is caused by shedding of active tissue factor from dissected atherosclerotic plaque. AB - Defined angiographically, no-reflow (NR) manifests as an acute reduction in coronary flow in the absence of epicardial vessel obstruction. One candidate protein to cause coronary NR is tissue factor (TF), which is abundant in atherosclerotic plaque and a cofactor for activated plasma coagulation factor VII. Scrapings from atherosclerotic carotid arteries contained TF activity (corresponding to 33.03 +/- 13.00 pg/cm(2) luminal plaque surface). Active TF was sedimented, indicating that TF was associated with membranes. Coronary blood was drawn from 6 patients undergoing coronary interventions with the distal protection device PercuSurge GuardWire (Traatek, Miami, FL). Fine particulate material that was recovered from coronary blood showed TF activity (corresponding to 91.1 +/- 62.16 pg/mL authentic TF). To examine the role of TF in acute coronary NR, blood was drawn via a catheter from coronary vessels in 13 patients during NR and after restoration of flow. Mean TF antigen levels were elevated during NR (194.3 +/- 142.8 pg/mL) as compared with levels after flow restoration (73.27 +/- 31.90 pg/mL; P =.02). To dissect the effects of particulate material and purified TF on flow, selective intracoronary injection of atherosclerotic material or purified relipidated TF was performed in a porcine model. TF induced NR in the model, thus strengthening the concept that TF is causal, not just a bystander to atherosclerotic plaque material. The data suggest that active TF is released from dissected coronary atherosclerotic plaque and is one of the factors causing the NR phenomenon. Thus, blood-borne TF in the coronary circulation is a major determinant of flow. PMID- 11929769 TI - Differential catalytic properties and vascular topography of murine nucleoside triphosphate diphosphohydrolase 1 (NTPDase1) and NTPDase2 have implications for thromboregulation. AB - Nucleoside triphosphate diphosphohydrolases (NTPDases) are a recently described family of ectonucleotidases that differentially hydrolyze the gamma and beta phosphate residues of extracellular nucleotides. Expression of this enzymatic activity has the potential to influence nucleotide P2 receptor signaling within the vasculature. We and others have documented that NTPDase1 (CD39, 78 kd) hydrolyzes both triphosphonucleosides and diphosphonucleosides and thereby terminates platelet aggregation responses to adenosine diphosphate (ADP). In contrast, we now show that NTPDase2 (CD39L1, 75 kd), a preferential nucleoside triphosphatase, activates platelet aggregation by converting adenosine triphosphate (ATP) to ADP, the specific agonist of P2Y(1) and P2Y(12) receptors. We developed specific antibodies to murine NTPDase1 and NTPDase2 and observed that both enzymes are present in the cardiac vasculature; NTPDase1 is expressed by endothelium, endocardium, and to a lesser extent by vascular smooth muscle, while NTPDase2 is associated with the adventitia of muscularized vessels, microvascular pericytes, and other cell populations in the subendocardial space. Moreover, NTPDase2 represents a novel marker for microvascular pericytes. Differential expression of NTPDases in the vasculature suggests spatial regulation of nucleotide-mediated signaling. In this context, NTPDase1 should abrogate platelet aggregation and recruitment in intact vessels by the conversion of ADP to adenosine monophosphate, while NTPDase2 expression would promote platelet microthrombus formation at sites of extravasation following vessel injury. Our data suggest that specific NTPDases, in tandem with ecto-5' nucleotidase, not only terminate P2 receptor activation and trigger adenosine receptors but may also allow preferential activation of specific subsets of P2 receptors sensitive to ADP (e.g., P2Y(1), P2Y(3), P2Y(12)) and uridine diphosphate (P2Y(6)). PMID- 11929770 TI - Plasminogen activator inhibitor type 2: a regulator of monocyte proliferation and differentiation. AB - We have explored the role of plasminogen activator inhibitor type 2 (PAI-2) in THP-1 monocyte-like cells. These cells possess a mutation in the PAI-2 gene and do not produce an active PAI-2 protein. Transfection of THP-1 cells with plasmids expressing active PAI-2 reduced the cells' inherent adhesive properties and decreased the rate of cell proliferation. THP-1 cells expressing active PAI-2 also displayed an altered phenotype in response to phorbol ester-induced differentiation that was concomitant with a reduction in CD14 expression. THP-1 cells transfected with a variant PAI-2 containing a mutation in the reactive center (PAI-2(Ala380)) displayed no noticeable change in any of these parameters, suggesting the involvement of a PAI-2-sensitive serine protease(s). The antiproliferative effect of PAI-2 was attenuated by treating the PAI-2-expressing THP-1 cells with recombinant urokinase (u-PA), suggesting that PAI-2 was disruptive of a u-PA/u-PA receptor signaling pathway initiated on the cell surface. Consistent with this, treatment of wild-type THP-1 cells with recombinant PAI-2 also caused a reduction in cellular proliferation. These results implicate endogenous PAI-2 as a modulator of monocyte adhesion, proliferation, and differentiation. PMID- 11929771 TI - Agonist-induced aggregation of Chinese hamster ovary cells coexpressing the human receptors for fibrinogen (integrin alphaIIbbeta3) and the platelet-activating factor: dissociation between adhesion and aggregation. AB - This work reports the establishment of a Chinese hamster ovary (CHO) cell line stably coexpressing the human alphaIIbbeta3 integrin and the platelet-activating factor receptor (PAFR). These cells aggregate in response to PAF in a Ca(++), alphaIIbbeta3, and soluble fibrinogen (Fg)-dependent manner that is prevented by PAF antagonists or alphaIIbbeta3 blockade. The aggregating response is accompanied by enhanced binding of fibrinogen and the activation-dependent IgM PAC1. This model has permitted us to identify, for the first time, intracellular signals distinctly associated with either alphaIIbbeta3-mediated adhesion or aggregation. Nonreceptor activation of protein kinase C (PKC) by phorbol ester produced cellular adhesion and spreading onto immobilized Fg, but it was not a sufficient signal to provoke cellular aggregation. Moreover, inhibition of PKC impeded the PAF stimulation of cellular adhesion, whereas the aggregation was not prevented. The PAF-induced cellular aggregation was distinctly associated with signaling events arising from the liganded Fg receptor and the agonist-induced stimulation of a calcium/calmodulin-dependent signaling pathway. Sustained tyrosine phosphorylation of both mitogen-activated protein kinase (MAPK) and an approximately 100-kd protein was associated with the PAF-induced aggregation, whereas phosphorylation of focal adhesion kinase (FAK) was preferably associated with cellular adherence and spreading onto immobilized Fg. PMID- 11929772 TI - Two classes of germline genes both derived from the V(H)1 family direct the formation of human antibodies that recognize distinct antigenic sites in the C2 domain of factor VIII. AB - Most plasmas from patients with inhibitors contain antibodies that are reactive with the C2 domain of factor VIII. Previously, we have shown that the variable heavy chain (V(H)) regions of antibodies to the C2 domain are encoded by the closely related germline gene segments DP-10, DP-14, and DP-88, which all belong to the V(H)1 gene family. Here, we report on the isolation and characterization of additional anti-C2 antibodies that are derived from V(H) gene segments DP-88 and DP-5. Competition experiments using murine monoclonal antibodies CLB-CAg 117 and ESH4 demonstrated that antibodies derived from DP-5 and DP-88 bound to different sites within the C2 domain. Epitope mapping studies using a series of factor VIII/factor V hybrids revealed that residues 2223 to 2332 of factor VIII are required for binding of the DP-10-, DP-14-, and DP-88-encoded antibodies. In contrast, binding of the DP-5-encoded antibodies required residues in both the amino- and carboxy-terminus of the C2 domain. Inspection of the reactivity of the antibodies with a series of human/porcine hybrids yielded similar data. Binding of antibodies derived from germline gene segments DP-10, DP-14, and DP-88 was unaffected by replacement of residues 2181 to 2243 of human factor VIII for the porcine sequence, whereas binding of the DP-5-encoded antibodies was abrogated by this replacement. Together these data indicate that antibodies assembled from V(H) gene segments DP-5 and the closely related germline gene segments DP-10, DP 14, and DP-88 target 2 distinct antigenic sites in the C2 domain of factor VIII. PMID- 11929773 TI - Plasmin activity is required for myogenesis in vitro and skeletal muscle regeneration in vivo. AB - Plasmin, the primary fibrinolytic enzyme, has a broad substrate spectrum and is implicated in biologic processes dependent upon proteolytic activity, such as tissue remodeling and cell migration. Active plasmin is generated from proteolytic cleavage of the zymogen plasminogen (Plg) by urokinase-type plasminogen activator (uPA) and tissue-type plasminogen activator (tPA). Here, we have investigated the role of plasmin in C2C12 myoblast fusion and differentiation in vitro, as well as in skeletal muscle regeneration in vivo, in wild-type and Plg-deficient mice. Wild-type mice completely repaired experimentally damaged skeletal muscle. In contrast, Plg(-/-) mice presented a severe regeneration defect with decreased recruitment of blood-derived monocytes and lymphocytes to the site of injury and persistent myotube degeneration. In addition, Plg-deficient mice accumulated fibrin in the degenerating muscle fibers; however, fibrinogen depletion of Plg-deficient mice resulted in a correction of the muscular regeneration defect. Because we found that uPA, but not tPA, was induced in skeletal muscle regeneration, and persistent fibrin deposition was also reproducible in uPA-deficient mice following injury, we propose that fibrinolysis by uPA-dependent plasmin activity plays a fundamental role in skeletal muscle regeneration. In summary, we identify plasmin as a critical component of the mammalian skeletal muscle regeneration process, possibly by preventing intramuscular fibrin accumulation and by contributing to the adequate inflammatory response after injury. Finally, we found that inhibition of plasmin activity with alpha2-antiplasmin resulted in decreased myoblast fusion and differentiation in vitro. Altogether, these studies demonstrate the requirement of plasmin during myogenesis in vitro and muscle regeneration in vivo. PMID- 11929774 TI - Flt3 ligand as a vaccine adjuvant in association with HER-2/neu peptide-based vaccines in patients with HER-2/neu-overexpressing cancers. AB - Dendritic cells (DCs) are potent antigen-presenting cells and have shown promise to function as "natural" vaccine adjuvants. Currently, most cancer vaccine trials using DCs generate autologous DCs ex vivo for each patient. Systemic treatment with Flt3 ligand (FL) results in a marked increase of DCs in tissues such as spleen and lymph nodes in mice and in the peripheral blood and skin of humans. In light of these observations, we questioned whether FL could be used systemically as a vaccine adjuvant to stimulate DC mobilization in vivo, circumventing the need to generate DCs ex vivo. Ten patients with HER-2/neu-overexpressing cancer were enrolled in a phase 1 study to receive a HER-2/neu peptide-based vaccine targeting the intracellular domain of the HER-2/neu protein. All patients received 20 microg/kg FL per day subcutaneously for 14 days. Five patients received the HER-2/neu peptide-based vaccine alone on day 7 of the 14-day cycle, and 5 patients received the vaccine admixed with 150 microg granulocyte macrophage-colony-stimulating factor (GM-CSF) on day 7 of the FL cycle. T-cell proliferative responses to HER-2/neu peptides and intracellular domain protein suggest that vaccine regimens including FL as an adjuvant were not effective in eliciting a significant HER-2/neu protein-specific T-cell proliferative response. However, including FL as a vaccine adjuvant was effective in boosting the precursor frequency of interferon-gamma-secreting HER-2/neu-specific T cells. The small sample size of each group, however, did not allow a statistically significant comparison of immune responses between the FL alone and FL with GM CSF arms. Finally, vaccine regimens including FL as a vaccine adjuvant were associated with the development of apparent autoimmune phenomena in some patients. PMID- 11929775 TI - Effects of exogenous interleukin-7 on human thymus function. AB - Immune reconstitution is a critical component of recovery after treatment of human immunodeficiency virus (HIV) infection, cancer chemotherapy, and hematopoietic stem cell transplantation. The ability to enhance T-cell production would benefit such treatment. We examined the effects of exogenous interleukin-7 (IL-7) on apoptosis, proliferation, and the generation of T-cell receptor rearrangement excision circles (TRECs) in human thymus. Quantitative polymerase chain reaction demonstrated that the highest level of TRECs (14 692 copies/10 000 cells) was present in the CD1a(+)CD3(-)CD4(+)CD8(+) stage in native thymus, suggesting that TREC generation occurred following the cellular division in this subpopulation. In a thymic organ culture system, exogenous IL-7 increased the TREC frequency in fetal as well as infant thymus, indicating increased T-cell receptor (TCR) rearrangement. Although this increase could be due to the effect of IL-7 to increase thymocyte proliferation and decrease apoptosis of immature CD3(-) cells, the in vivo experiments using NOD/LtSz-scid mice given transplants of human fetal thymus and liver suggested that IL-7 can also directly enhance TREC generation. Our results provide compelling evidence that IL-7 has a direct effect on increasing TCR-alphabeta rearrangement and indicate the potential use of IL-7 for enhancing de novo naive T-cell generation in immunocompromised patients. PMID- 11929776 TI - Disrupted homeostasis of Langerhans cells and interdigitating dendritic cells in monkeys with AIDS. AB - Langerhans cells (LCs) are immature dendritic cells (DCs) that capture antigen in peripheral tissues and migrate to draining lymph nodes, where they reside in the paracortex as interdigitating dendritic cells (IDCs). We studied the effects of simian immunodeficiency virus (SIV) on LCs and IDCs during different stages of infection in monkeys. LCs isolated from monkeys with acute SIV infection or acquired immunodeficiency syndrome (AIDS) underwent normal maturation in vitro, including a switch in chemokine receptor expression from CCR5 to CXCR4 and CCR7. LCs migrated normally from skin in response to contact sensitization in monkeys with acute SIV infection. In contrast, LC migration from skin was markedly impaired during AIDS, associated with a reduction in antigen-bearing DCs in draining lymph nodes. Lymph node IDCs were increased in proportion during acute SIV infection and had an activated phenotype, whereas during AIDS IDCs had significantly lower expression of CD40 and the activation marker CD83. IDCs from monkeys with AIDS were refractory to stimulation with CD40L, demonstrating a functional consequence of decreased CD40 expression. SIV-infected DCs were not identified in lymph nodes or skin of monkeys with AIDS, suggesting an indirect effect of infection on DC populations in vivo. These data indicate that DCs are mobilized to lymph nodes during acute SIV infection, but that during AIDS this process is suppressed, with LC migration and IDC activation being impaired. We conclude that disruption of DC homeostasis may play a role in immunopathology induced by human immunodeficiency virus and suggest that therapeutic strategies targeting DCs may have limited efficacy during AIDS. PMID- 11929777 TI - Increased dendritic cell number and function following continuous in vivo infusion of granulocyte macrophage-colony-stimulating factor and interleukin-4. AB - Dendritic cells (DCs) are rare antigen-presenting cells that play a central role in stimulating immune responses. The combination of recombinant granulocyte macrophage-colony-stimulating factor (rGM-CSF) and recombinant interleukin-4 (rIL 4) provides an important stimulus for generating DCs from murine bone marrow precursors in vitro. Using miniature osmotic pumps, we now demonstrate that continuous infusion of these cytokines for 7 days had a similar effect in vivo, increasing the number and function of splenic DCs. Administration of rGM-CSF/rIL 4 (10 microg/d each) increased the concentration of CD11(+) DCs by 2.7-fold and the absolute number of splenic DCs by an average of 5.7-fold. DC number also increased in peripheral blood and lymph nodes. The resultant DCs exhibited a different phenotype and function than those in control mice or mice treated with rGM-CSF alone. rGM-CSF/IL-4 increased both the myeloid (CD11c(+)/CD11b(+)) and the lymphoid (CD11c(+)/CD8alpha(+)) subpopulations, whereas rGM-CSF increased only myeloid DCs. DCs were highly concentrated in the T-cell areas of white pulp after rGM-CSF/IL-4 administration, whereas they were diffusely distributed throughout white pulp, marginal zones, and red pulp in mice treated with rGM-CSF alone. rGM-CSF/rIL-4 also significantly increased the expression of major histocompatibility complex (MHC) class I and MHC class II on CD11c(+) cells and increased their capacity to take up antigens by macropinocytosis and receptor mediated endocytosis. Splenic DCs generated in response to rGM-CSF/rIL-4 were functionally immature in terms of allostimulatory activity, but this activity increased after short-term in vitro culture. Systemic treatment with rGM-CSF/rIL 4 enhanced the response to an adenoviral-based vaccine and led to antigen specific retardation in the growth of established tumor. We conclude that systemic therapy with the combination of rGM-CSF/rIL-4 provides a new approach for generating DCs in vivo. PMID- 11929778 TI - B7-CTLA4 interaction promotes cognate destruction of tumor cells by cytotoxic T lymphocytes in vivo. AB - Costimulatory molecules B7-1 and B7-2 (hereby collectively called B7) interact with CD28 and CTLA4 on T cells and promote antitumor immunity. The function of B7 CTLA4 interaction in antitumor CTL response remains controversial. Here we used CD28(-/-) and CD28(+/-) or CD28(+/+) transgenic mice that express the T-cell receptor specific for an unmutated tumor antigen, P1A, and for tumor cells expressing a CTLA4-specific B7 mutant to evaluate the function of CD28-B7 and CTLA4-B7 interactions in induction and effector phases of antitumor immunity. We report that B7-CD28 and B7-CTLA4 interactions promote tumor rejection. However, this is achieved by distinct mechanisms. B7-CD28 interaction enhances T-cell clonal expansion, though a role for this interaction in the effector phase cannot be ruled out. In contrast, B7-CTLA4 interaction enhances the CTL-mediated destruction of tumors, but not T-cell clonal expansion. PMID- 11929779 TI - Human mast cell progenitors use alpha4-integrin, VCAM-1, and PSGL-1 E-selectin for adhesive interactions with human vascular endothelium under flow conditions. AB - Mast cells (MCs) are central to asthma and other allergic diseases, and for responses to infection and tissue injuries. MCs arise from committed progenitors (PrMCs) that migrate from the circulation to tissues by incompletely characterized mechanisms, and differentiate in situ in perivascular connective tissues of multiple organs. PrMCs derived in vitro from human cord blood were examined for adhesion molecule expression and their ability to adhere to human umbilical vein endothelial cells (HUVECs) under conditions that mimic physiologic shear flow. The PrMCs expressed alpha(4)beta(1), low levels of beta7, and the beta2-integrins alphaLbeta2 and alphaMbeta2. The PrMCs also expressed PSGL-1, but not L-selectin. At low (0.5 dynes/cm(2)-1.0 dynes/cm(2)) shear stress, PrMCs attached and rolled on recombinant E-selectin and P-selectin and VCAM-1. An anti PSGL-1 monoclonal antibody (mAb) blocked essentially all adhesion to P-selectin but reduced adhesion to E-selectin by only 40%, suggesting PrMCs express other ligands for E-selectin. PrMCs adhered strongly to tumor necrosis factor-alpha (TNF-alpha)-activated HUVECs, whereas adhesion to interleukin 4 (IL-4)-activated HUVECs was lower. PrMC adhesion to IL-4-activated HUVECs was totally alpha4 integrin- and VCAM-1-dependent. Adhesion to TNF-alpha-activated HUVECs was blocked by 50% by mAbs against alpha4-integrin, vascular cell adhesion molecule-1 (VCAM-1), E-selectin, or PSGL-1, whereas combinations of mAbs to alpha4-integrin plus PSGL-1, or VCAM-1 plus E-selectin, blocked adhesion by greater than 70%. Thus, PrMCs derived in vitro predominantly use alpha4-integrin, VCAM-1, PSGL-1, and other ligands that bind E-selectin for adhesion to cytokine-activated HUVEC monolayers. These observations may explain the abundance of MCs at sites of mucosal inflammation, where VCAM-1 and E-selectin are important inducible receptors. PMID- 11929780 TI - Spontaneous generation and survival of blood dendritic cells in mononuclear cell culture without exogenous cytokines. AB - Studies on purified blood dendritic cells (DCs) are hampered by poor viability in tissue culture. We, therefore, attempted to study some of the interactions/relationships between DCs and other blood cells by culturing unseparated peripheral blood mononuclear cell (PBMC) preparations in vitro. Flow cytometric techniques were used to undertake a phenotypic and functional analysis of DCs within the cultured PBMC population. We discovered that both the CD11c(+) and CD11c(-) CD123(hi) DC subsets maintained their viability throughout the 3-day culture period, without the addition of exogenous cytokines. This viability was accompanied by progressive up-regulation of the surface costimulatory (CD40, CD80, CD86) and activation (CMRF-44, CMRF-56, CD83) molecules. The survival and apparent production of DCs in PBMC culture (without exogenous cytokines) and that of sorted DCs (with cytokines) were evaluated and compared by using TruCOUNT analysis. Absolute DC counts increased (for CD123(hi) and CD11c(+) subsets) after overnight culture of PBMCs. Single-cell lineage depletion experiments demonstrated the rapid and spontaneous emergence of "new" in vitro generated DCs from CD14(+)/CD16(+) PBMC radioresistant precursors, additional to the preexisting ex vivo DC population. Unlike monocyte-derived DCs, blood DCs increased dextran uptake with culture and activation. Finally, DCs obtained after culture of PBMCs for 3 days were as effective as freshly isolated DCs in stimulating an allogeneic mixed leukocyte reaction. PMID- 11929781 TI - IgG plasma cells display a unique spectrum of leukocyte adhesion and homing molecules. AB - Long-lived antibody-secreting plasma cells are formed in the secondary lymphoid organs and subsequently home to the bone marrow, although the mechanisms that control this migration remain primarily unknown. In this study, we show that IgG plasma cells constitute a significant fraction of cervical lymph node cells from older mice deficient in both E- and P-selectin (E/P(-/-)), and that these cells can be prospectively isolated by phenotype. These IgG plasma cells were polyclonal, cytoplasmic Ig(+), spontaneously secreted antibody, were in the G(0)/G(1) phase of the cell cycle, and failed to express multiple B-cell surface markers. The plasma cells exhibited up-regulated cell surface expression of multiple adhesion molecules, including alpha(4) and leukocyte function-associated antigen 1 (LFA-1) integrins, CD44, and P-selectin glycoprotein ligand 1 (PSGL-1). IgG plasma cells bound to vascular cell adhesion molecule 1 (VCAM-1) significantly better than IgM(+) B cells, indicating that the alpha(4) integrins were constitutively active. A subset of IgG plasma cells also bound hyaluronic acid, the ligand for CD44. In addition, the IgG plasma cells interacted strongly with E-selectin, but poorly with P-selectin, despite elevated levels of PSGL-1 protein. The preferential interaction of plasma cells with E-selectin, but not P selectin, correlated with elevated alpha1,3-fucosyltransferase-VII messenger RNA levels, but selective down-regulation of core 2 beta1-6-N-glucosaminyltransferase levels, compared to B cells. These results demonstrate a unique adhesion profile for murine IgG plasma cells. Furthermore, the E/P(-/-) mice represent a novel system to isolate and purify significant numbers of primary IgG plasma cells. PMID- 11929782 TI - Human cytomegalovirus inhibits maturation and impairs function of monocyte derived dendritic cells. AB - Dendritic cells (DCs) play a pivotal role in the generation of virus-specific cytotoxic T-cell responses, but some viruses can render DCs inefficient in stimulating T cells. We studied whether infection of DCs with human cytomegalovirus (HCMV) results in a suppression of DC function which may assist HCMV in establishing persistence. The effect of HCMV infection on the phenotype and function of monocyte-derived DCs and on their ability to mature following infection with an endothelial cell-adapted clinical HCMV isolate were studied. HCMV infection induced no maturation of DCs; instead, it efficiently down regulated the expression of surface major histocompatibility complex (MHC) class I, CD40, and CD80 molecules. Slight down-regulation of MHC class II and CD86 molecules was also observed. Lipopolysaccharide (LPS)-induced maturation of infected DCs was strongly inhibited, as indicated by lower levels of surface expression of MHC class I, class II, costimulatory, and CD83 molecules. The down regulation or inhibition of these surface markers occurred only in HCMV antigen positive DCs. DCs produced no interleukin 12 (IL-12) and only low levels of tumor necrosis factor alpha (TNF-alpha) upon HCMV infection. Furthermore, cytokine production upon stimulation with LPS or CD40L was significantly impaired. Inhibition of cytokine production did not depend on viral gene expression as UV irradiated HCMV resulted in the same effect. Proliferation and cytotoxicity of T cells specific to a recall antigen presented by DCs were also reduced when DCs were HCMV infected. This study shows that HCMV inhibits DC function, revealing a powerful viral strategy to delay or prevent the generation of virus-specific cytotoxic T cells. PMID- 11929783 TI - Level of galactosylation determines cryoglobulin activity of murine IgG3 monoclonal rheumatoid factor. AB - Autoantibodies of the cryoprecipitating IgG3 isotype have been shown to play a significant role in the development of murine lupus-like autoimmune syndrome. At present, the structural basis of IgG3 cryoprecipitation and its role in autoantibody pathogenicity remain to be defined. Using molecular variants of an IgG3 monoclonal rheumatoid factor, 6-19, derived from an autoimmune MRL-Fas(lpr) mouse, we have investigated the implication of charged residues in the heavy chain variable (VH) region, potential CH3-linked oligosaccharides, and galactosylation of CH2-linked oligosaccharides in its cryoglobulin activity. The cryoglobulin activity of the IgG3 6-19 mutant bearing more negatively charged residues at VH 6 and 23 was found to be reduced but still highly significant, whereas that of the mutant lacking a potential CH3 glycosylation site remained unchanged. In marked contrast, IgG3 6-19 variants obtained from 6-19 heavy-chain transgenic mice displayed barely detectable cryoglobulin activity associated with an increased level of galactosylation in the CH2 oligosaccharide side chains. Thus, our data strongly suggest that the cryoglobulin activity of IgG3 6-19 autoantibody is critically determined by levels of galactosylation in the CH2 oligosaccharide side chains, whereas VH residues play a secondary role in 6-19 IgG3 cryoglobulin activity. PMID- 11929784 TI - The growth of cutaneous T-cell lymphoma is stimulated by immature dendritic cells. AB - In the initial stage of cutaneous T-cell lymphoma (CTCL), proliferating CTCL cells are concentrated in the epidermis in close association with an immature dendritic cell (DC), the Langerhans cell. Because long-term in vitro culture of CTCL cells has proven difficult, the in vivo association with the major antigen presenting cell (APC) of the epidermis has been postulated to play a role in directly stimulating the clonal T-cell proliferation. We report that CTCL cells can be reproducibly grown in culture for 3 months when cocultured with immature DCs. CTCL cells retain the phenotype and genotype of the initial malignant clone, whereas the APCs are a mixture of immature and mature DCs. CTCL cell and DC survival was dependent on direct membrane contact. Growth was inhibited by antibodies that bound to the T-cell receptor (TCR) or interfered with the interaction of CD40 with its ligand on the CTCL cell. Addition of antibody to CD3 or the clonotypic TCR caused rapid CTCL cell apoptosis followed by engulfment by avidly phagocytic immature DCs and subsequent DC maturation. The opportunity to study CTCL cells and immature DCs for prolonged periods will facilitate studies of tumor cell biology and will allow investigation of the intriguing hypothesis that CTCL cell growth is driven through TCR recognition of class II-presented self-peptides. In addition, the culture of CTCL cells will permit evaluation of therapies in vitro before clinical intervention, thereby improving safety and efficacy. PMID- 11929785 TI - CD95 engagement induces disseminated endothelial cell apoptosis in vivo: immunopathologic implications. AB - Fas (CD95) is a death receptor involved in apoptosis induction on engagement by Fas ligand (CD95L). Although CD95L-mediated apoptosis has been proposed as a pathogenic mechanism in a wide range of diseases, including graft-versus-host disease, systemic CD95 engagement in mice by agonistic CD95-specific antibodies or by soluble multimeric CD95L (smCD95L), though lethal, has been reported to cause apoptosis only in a limited range of cell types, that is, hepatocytes, hepatic sinusoidal endothelial cells, and lymphocytes. Another member of the tumor necrosis factor (TNF)/CD95L family, TNF-alpha, induces disseminated vascular endothelial cell apoptosis, which precedes apoptosis of other cell types and lethal multiorgan failure. Here we show that systemic CD95 engagement in vivo by agonistic CD95-specific antibody or smCD95L causes rapid, extensive, and disseminated endothelial cell apoptosis throughout the body, by a mechanism that does not depend on TNF-alpha. Disseminated endothelial cell apoptosis was also the first detectable lesion in a murine model of acute tissue damage induced by systemic transfer of allogeneic lymphocytes and did not occur when allogeneic lymphocytes were from CD95L-defective mice. Both vascular and additional tissue lesions induced by agonistic CD95-specific antibody, smCD95L, or allogeneic lymphocytes were prevented by treatment with an inhibitor of caspase-8, the upstream caspase coupled to CD95 death signaling. Vascular lesions are likely to play an important role in the pathogenesis of allogeneic immune responses and of other diseases involving circulating CD95L-expressing cells or smCD95L, and the prevention of CD95-mediated death signaling in endothelial cells may have therapeutic implications in these diseases. PMID- 11929786 TI - Identification of a myeloid intrathymic pathway of dendritic cell development marked by expression of the granulocyte macrophage-colony-stimulating factor receptor. AB - In this study, the finding that a significant proportion of all dendritic cells (DCs) resident in vivo in the human postnatal thymus displayed a myeloid-related phenotype prompted us to re-examine the developmental origin of thymic DCs, a cell type hitherto considered to represent a homogeneous lymphoid-derived population. We show here that these novel intrathymic DCs are truly myeloid, as they arise from CD34(+) early thymic progenitors through CD34(lo) intermediates which have lost the capacity to generate T cells, but display myelomonocytic differentiation potential. We also demonstrate that phenotypically and functionally equivalent myeloid precursors devoid of T-cell potential do exist in vivo in the postnatal thymus. Moreover, although interleukin 7 (IL-7) supports the generation of such myeloid intermediates, we show that their developmental branching from the main intrathymic T-cell pathway is linked to the up-regulation of the myelomonocytic granulocyte macrophage-colony-stimulating factor (GM-CSF) receptor, to the down-regulation of the IL-7 receptor and to the lack of pre-T cell receptor alpha (pTalpha) gene transcriptional activation. Taken together, these data challenge the current view that the thymus is colonized by a lymphoid restricted progenitor and provide evidence that a more immature precursor population with lymphoid and myelomonocytic potential is actually seeding the human postnatal thymus. PMID- 11929787 TI - The coiled-coil domain and Tyr177 of bcr are required to induce a murine chronic myelogenous leukemia-like disease by bcr/abl. AB - The bcr/abl fusion in chronic myelogenous leukemia (CML) creates a chimeric tyrosine kinase with dramatically different properties than intact c-abl. In P210 bcr/abl, the bcr portion includes a coiled-coil oligomerization domain (amino acids 1-63) and a grb2-binding site at tyrosine 177 (Tyr177) that are critical for fibroblast transformation, but give variable results in other cell lines. To investigate the role of the coiled-coil domain and Tyr177 in promoting CML, 4 P210 bcr/abl-derived mutants containing different bcr domains fused to abl were constructed. All 4 mutants, Delta(1-63) bcr/abl, (1-63) bcr/abl, Tyr177Phe bcr/abl, and (1-210) bcr/abl exhibited elevated tyrosine kinase activity and conferred factor-independent growth in cell lines. In contrast, differences in the transforming potential of the 4 mutants occurred in our mouse model, in which all mice receiving P210 bcr/abl-expressing bone marrow cells exclusively develop a myeloproliferative disease (MPD) resembling human CML. Of the 4 mutants assayed, only 1-210 bcr/abl, containing both the coiled-coil domain and Tyr177, induced MPD. Unlike full-length P210, this mutant also caused a simultaneous B cell acute lymphocytic leukemia (ALL). The other 3 mutants, (1-63) bcr/abl, Tyr177Phe bcr/abl, and Delta(1-63) bcr/abl, failed to induce an MPD but instead caused T-cell ALL. These results show that both the bcr coiled-coil domain and Tyr177 are required for MPD induction by bcr/abl and provide the basis for investigating downstream signaling pathways that lead to CML. PMID- 11929788 TI - Involvement of protein kinase C and phosphatidylinositol 3-kinase pathways in the survival of B-cell chronic lymphocytic leukemia cells. AB - B-cell chronic lymphocytic leukemia (B-CLL) is characterized by the accumulation of long-lived CD5(+) B lymphocytes. TPA (12-O-tetradecanoylphorbol 13- acetate) and interleukin-4 (IL-4) inhibit apoptosis of B-CLL lymphocytes ex vivo. We used specific inhibitors of protein kinase C (PKC), extracellular-regulated kinase (ERK), and phosphatidylinositol 3-kinase (PI3-kinase) to study their involvement in TPA- and IL-4-induced survival of B-CLL lymphocytes. BisI, a specific inhibitor of PKC, induced apoptosis and inhibited the antiapoptotic activity of TPA and IL-4. B-CLL cells have a basal PKC activity that was increased by TPA but not by IL-4. TPA, but not IL-4, induced ERK activation. However, the inhibition of ERK activation did not affect the viability of B-CLL lymphocytes, demonstrating that this pathway is not involved in their survival. Inhibition of PI3-kinase by LY294002 induced apoptosis of B-CLL cells and inhibited the survival effect of IL-4 and TPA. In addition, Akt, a downstream effector of PI3 kinase activity, was phosphorylated by TPA and IL-4 in B-CLL cells, though PI3 kinase had no effect on PKC-dependent phosphorylation of Akt. Furthermore, the inhibition of PKC or PI3-kinase increased dexamethasone- and fludarabine-induced apoptosis ex vivo in the presence of survival factors. These results demonstrate that PKC and PI3-kinase are involved in the survival of B-CLL cells and suggest that inhibitors of these pathways could be combined with the drugs used in the treatment of B-CLL. PMID- 11929789 TI - The chemokine receptor CCR7 and alpha4 integrin are important for migration of chronic lymphocytic leukemia cells into lymph nodes. AB - Malignant lymphocyte migration into lymph nodes is an important aspect of chronic lymphocytic leukemia (CLL), yet little is known about the processes involved. Here we demonstrate that CLL cells migrate across vascular endothelium in response to at least 3 chemokines, namely, CCL21, CCL19, and CXCL12. Moreover, transendothelial cell migration (TEM) in response to CCL21 and CCL19 was significantly higher for the malignant B cells of patients who had clinical lymph node involvement as compared with those of patients lacking such organomegaly. Furthermore, the expression of CCR7, the receptor for both CCL21 and CCL19, correlated with clinical lymphadenopathy, and blocking of CCR7 inhibited CLL cell TEM. By using immunohistochemistry we demonstrated that CCL21 and CCL19, but not CXCL12, are located in high endothelial venules and are, therefore, in an appropriate location to induce TEM. Regarding the adhesion receptors involved in TEM, alpha4 (most likely in association with beta1) and alphaLbeta2 were shown to be important in CLL cell TEM in vitro, but only the level of alpha4 expression correlated with the presence of clinical lymphadenopathy. The present studies are the first to shed light on the factors determining CLL cell entry into nodes and define the phenotype of circulating malignant cells likely to determine the pattern of lymph node enlargement in the disease. PMID- 11929790 TI - Recurring chromosomal abnormalities in leukemia in PML-RARA transgenic mice parallel human acute promyelocytic leukemia. AB - Acute promyelocytic leukemia (APL) is characterized by the t(15;17)(q22;q11.2), which results in the PML-RARA fusion gene. In previous studies, we demonstrated that expression of a human PML-RARA complementary DNA in murine granulocyte precursor cells initiated the development of leukemia. However, leukemogenesis by PML-RARA required additional genetic alterations. To identify genetic changes that cooperate with PML-RARA in leukemogenesis, we performed spectral karyotyping analysis of myeloid leukemias from hMRP8-PML-RARA mice (11 cases) and from mice coexpressing PML-RARA and BCL2 (8 cases). Clonal abnormalities were detected in 18 of 19 cases (95%). Recurring numerical abnormalities identified in these murine leukemias included +15 (15 cases, 79%); loss of a sex chromosome (12 cases, 63%); +8 (10 cases, 53%); +10 (9 cases, 47%); +4, +7, or +14 (8 cases each, 42%); +16 (7 cases, 37%); and +6 (5 cases, 26%). In a series of 965 patients with APL, we identified secondary abnormalities in 368 (38%). The most common recurring abnormalities were +8 or partial trisomy of 8q (120 patients, 12.4%) and ider(17) t(15;17) (42 patients, 4.4%). The critical consequence of +8 in human leukemias appears to be the gain of 8q24, which is syntenic to mouse 15. Thus, our results suggest that PML-RARA-initiated murine leukemia is associated with a defined spectrum of genetic changes, and that these secondary mutations recapitulate, in part, the cytogenetic abnormalities found in human APL. PMID- 11929791 TI - High frequency of leukemic clones in newborn screening blood samples of children with B-precursor acute lymphoblastic leukemia. AB - The detection of leukemia cells on newborn genetic screening cards ("Guthrie cards") of a small group of patients and several sets of identical twins developing acute lymphoblastic leukemia (ALL) with identical phenotypic and chromosomal markers has provided evidence that childhood ALL cases may arise in utero. We conducted a retrospective study of a randomly selected group of childhood B-precursor ALL patients to determine the frequency of the presence of "leukemic" clones prenatally in ALL cases by testing newborn screening cards. The 17 ALL patients analyzed had a median age of 46 months (range, 18 months to 13 years) and had median presenting white blood cell (WBC) counts of 10 950/microL (range, 2900-70 300/microL) at diagnosis. A clonal rearrangement of the immunoglobulin heavy chain (IgH) gene was identified in diagnostic lymphoblasts and sequenced and patient-specific primers were used to amplify DNA from blood samples on the patient's newborn screening cards. Twelve of the 17 (71%) analyzed newborn cards had detectable IgH rearrangements amplified by seminested polymerase chain reaction. DNA sequencing confirmed that the IgH rearrangements detected matched the IgH sequences identified from diagnostic leukemia cells, indicating the presence of a "leukemic" clone at birth. There were no differences in age or presenting WBC counts between the cases with or without positive newborn screening cards. All 6 patients with hyperdiploid ALL had detectable "leukemic" clones on their cards. The results of our study support the notion that a high proportion of childhood B-precursor ALL cases arise in utero, although postnatal events are also important factors in leukemogenesis. PMID- 11929792 TI - Presence of a mobilizable intracellular pool of hepatocyte growth factor in human polymorphonuclear neutrophils. AB - Hepatocyte growth factor (HGF), a heparin-binding factor, is synthesized as a single-chain inactive precursor (pro-HGF), which is converted by proteolysis to an active heterodimer (mature HGF). HGF has pleiotropic activities and has been implicated in the regulation of mitogenesis, motogenesis, and morphogenesis of epithelial and endothelial cells. As polymorphonuclear neutrophils (PMNs) secrete numerous cytokines involved in the modulation of local inflammation, we investigated their ability to produce HGF. We found that HGF was stored in secretory vesicles and in gelatinase/specific granules. This intracellular stock was rapidly mobilized by degranulation when neutrophils were stimulated with phorbol myristate acetate or N-formylmethionyl-leucyl-phenylalanine. Cycloheximide did not affect the release of HGF. Moreover, HGF messenger RNA and protein expression was found in bone marrow myeloid cells, suggesting that HGF synthesis likely occurs during PMN maturation. In mature circulating PMNs, intracellular HGF was in the pro-HGF form, whereas the HGF secreted by degranulation was the mature form. Furthermore, PMNs pretreated with diisopropyl fluorophosphate only released the pro-HGF form, suggesting that PMN-derived serine protease(s) are involved in the proteolytic process. We also obtained evidence that secreted mature HGF binds PMN-derived glycosaminoglycans (probably heparan sulfate). These findings suggest that PMNs infiltrating damaged tissues may modulate local wound healing and repair through the production of HGF, a major mediator of tissue regeneration. PMID- 11929793 TI - Fetal and adult hemoglobin production during adult erythropoiesis: coordinate expression correlates with cell proliferation. AB - The design and evaluation of therapies for the sickle cell and beta-thalassemia syndromes rely on our understanding of hemoglobin accumulation during human erythropoiesis. Here we report direct measurements of hemoglobin composition and messenger RNA (mRNA) levels in cultured CD34(+) cells and correlate those measurements with studies of freshly obtained bone marrow samples. Hemoglobin levels in differentiating cells were also compared with morphologic, immunophenotypic, and cell cycle assessments. A population of large preproerythroblasts was first identified within 24 hours and became the dominant population by day 5. The transition from proerythroblast to basophilic normoblast occurred later, from days 7 to 9, and correlated with a peak of 74.1% +/- 3.9% of the cells in the S phase of cell cycle. Orthochromatic normoblasts were the dominant and final cell type by day 13. High-performance liquid chromatography based quantitation of fetal (HbF) and adult (HbA) hemoglobin and real-time polymerase chain reaction globin mRNA quantitation demonstrated a coordinate rise in the accumulation of both proteins and mRNA among these developmentally staged populations. Quantitative analyses on freshly sorted bone marrow populations demonstrated a similar rising pattern with beta-globin and HbA as the dominant species at both early and late stages of differentiation. We found no evidence for HbF dominant populations or switching during differentiation in adult cells. Instead, rapid increases in both HbF (heterocellular) and HbA (pancellular) content were observed, which coincided with the apex in cell cycling and the proerythroblast-basophilic normoblast transition. Based on these measurements, we conclude that HbF and HbA content are regulated with the rate of proliferation during adult erythropoiesis. PMID- 11929794 TI - Longitudinal changes in brain magnetic resonance imaging findings in children with sickle cell disease. AB - Children with sickle cell anemia (HbSS) are at high risk for neurologically overt cerebral infarcts associated with stroke and neurologically silent cerebral infarcts correlated with neuropsychometric deficit. We used complete magnetic resonance imaging (MRI) histories from 266 HbSS children, aged 6 through 19 years, who were enrolled in the Cooperative Study of Sickle Cell Disease (CSSCD) to examine silent infarct prevalence, localization, recurrence, and progression. We report a baseline prevalence of 21.8%, marginally higher than previously reported due to improved imaging technologies. Although we observed no overall sex difference in prevalence, most lesions in girls occurred before age 6, whereas boys remained at risk until age 10. Silent infarcts were significantly smaller and less likely to be found in the frontal or parietal cortex than were infarcts associated with stroke. Children with silent infarct had an increased incidence of new stroke (1.03/100 patient-years) and new or more extensive silent infarct (7.06/100 patient-years) relative to stroke incidence among all children in our cohort (0.54/100 patient-years). Both events were substantially less frequent than the risk of stroke recurrence among children not provided chronic transfusion therapy. Although chronic transfusion is known to decrease occurrence of new silent infarcts and strokes in children with elevated cerebral arterial blood flow velocity, further study is required to determine its risk-benefit ratio in children with silent infarct and normal velocities. Until safe and effective preventive strategies against infarct recurrence are discovered, MRI studies are best reserved for children with neurologic symptoms, neuropsychometric deficits, or elevated cerebral artery velocities. PMID- 11929795 TI - HBED ligand: preclinical studies of a potential alternative to deferoxamine for treatment of chronic iron overload and acute iron poisoning. AB - We have continued the preclinical evaluation of the efficacy and safety of the hexadentate phenolic aminocarboxylate iron chelator N, N'-bis(2-hydroxybenzyl) ethylenediamine-N, N'-diacetic acid monosodium salt (NaHBED) for the treatment of both chronic transfusional iron overload and acute iron poisoning. We examined the effect of route of administration by giving equimolar amounts of NaHBED and deferoxamine (DFO) to Cebus apella monkeys as either a subcutaneous (SC) bolus or a 20-minute intravenous (IV) infusion. By both routes, NaHBED was consistently about twice as efficient as DFO in producing iron excretion. For both chelators at a dose of 150 micromol/kg, SC was more efficient than IV administration. The biochemical and histopathologic effects of NaHBED administration were assessed. No systemic toxicity was found after either IV administration once daily for 14 days to iron-loaded dogs or after SC administration every other day for 14 days to dogs without iron overload. Evidence of local irritation was found at some SC injection sites. When the NaHBED concentration was reduced to 15% or less in a volume comparable to a clinically useful one, no local irritation was found with SC administration in rats. Because treatment of acute iron poisoning may require rapid chelator infusion, we compared the effects of IV bolus administration of the compounds to normotensive rats. Administration of DFO produced a prompt, prolonged drop in blood pressure and acceleration of heart rate; NaHBED had little effect. NaHBED may provide an alternative to DFO for the treatment of both chronic transfusional iron overload and of acute iron poisoning. PMID- 11929797 TI - Activated eosinophils in upper gastrointestinal tract of patients with graft versus-host disease. AB - Digestive tract damage during graft-versus-host reaction (GVHR) causes high morbidity and mortality. Diagnosis is often late because biopsies are performed when clinical signs are severe and pathologic markers of early inflammatory lesions are lacking. Eosinophils are inflammatory cells, cytotoxic in vitro to digestive epithelium; they are found in biopsy specimens taken during acute flare ups of inflammatory bowel disease. We performed systematic duodenal biopsies immediately after digestive symptoms occurred and found a digestive GVHR incidence of 73.1% (n = 93), higher than that found when digestive biopsies were performed immediately after severe clinical signs. Eosinophils were only present when there were histologic signs of GVHR; eosinophil presence correlated with GVHR severity. Electron microscopy with immunogold staining showed pathologic signs of in situ eosinophil activation, such as cytoplasmic granule alterations, and eosinophil peroxidase release in all patients. Interleukin-5 presence in activated eosinophils suggests eosinophil recruitment in digestive GVHR is an autocrine mechanism. Eosinophil density also correlated with GVHR severity, whether in acute or chronic clinical phases. Tissue eosinophils could thus be a marker of acute inflammatory flare-ups in GVHR. Systematic duodenal biopsy performed at the onset of digestive symptoms should allow early GVHR detection, and pathologic signs of GVHR, together with eosinophil density, might help modulate immunosuppressive therapy. PMID- 11929796 TI - The DBY gene codes for an HLA-DQ5-restricted human male-specific minor histocompatibility antigen involved in graft-versus-host disease. AB - Graft rejection or graft-versus-host (GVH) disease after HLA-identical stem cell transplantation is the result of recognition of minor histocompatibility antigens (mHags) by immunocompetent T lymphocytes from recipient or donor origin, respectively. Cytolytic T lymphocyte (CTL) clones can be isolated during graft rejection and GVH disease to identify mHags and their corresponding genes. Thus far, all human mHags identified appeared to be HLA class I-restricted. Here, we report the characterization of the first human HLA class II-restricted sex-linked mHag involved in GVH disease. Previously, we isolated an HLA-DQ5-restricted CD4(+) CTL clone from a male patient with chronic myeloid leukemia who developed acute GVH disease grade III-IV after transplantation of HLA genotypically identical female stem cells. Using a panel of female HLA-DQ5(+) EBV cells that we stably transfected with Y chromosome-specific genes, we determined that the HLA class II male-specific mHag (H-Y) was encoded by the Y chromosome-specific gene DBY. The H-Y epitope was localized in the DBY protein using female HLA-DQ5(+) peripheral blood mononuclear cells loaded with DBY protein fragments. The minimal peptide sequence leading to maximal recognition by the specific HLA-DQ5 restricted CTL clone was characterized as the 12-amino acid sequence HIENFSDIDMGE. Although the epitope differed by 3 amino acids from its X-homolog DBX, only 2 polymorphisms were shown to be essential for recognition by the CTL clone. PMID- 11929798 TI - Murine acute graft-versus-host disease can be prevented by depletion of alloreactive T lymphocytes using activation-induced cell death. AB - Depletion of T lymphocytes from allogeneic bone marrow transplants successfully prevents the development of graft-versus-host disease (GvHD) but is associated with impaired engraftment, immunosuppression, and abrogation of the graft-versus leukemia effect. We therefore explored the possibility of selectively eliminating alloreactive T cells by CD95/CD95L-mediated activation-induced cell death (AICD) in a major histocompatibility complex allogeneic murine model system. Activation of resting or preactivated T lymphocytes from C3H/HeJ (H-2(k)) mice was induced with irradiated BALB/cJ (H-2(d)) mouse-derived stimulators. Substantial decrease (> or = 80%) of proliferative and lytic responses by activated alloreactive T cells was subsequently achieved by incubating the mixed lymphocyte culture with an agonistic monoclonal antibody to CD95, and residual T cells recovered did not elicit alloreactivity upon challenge to H-2(d). Depletion of alloreactive T lymphocytes by AICD was specific because reactivity to an I-A(d)-restricted ovalbumin (OVA) peptide by OVA-specific CD4(+) T cells mixed into the allogeneic T-cell pool and subjected to induction of AICD in the absence of OVA peptide could be preserved. Adoptive transfer of donor-derived allogeneic T lymphocytes, depleted from alloreactive T cells by AICD in vitro, in the parent (C3H/He) to F(1) (C3H/He x BALB/c) GvHD model prevented lethal GvHD. The results presented suggest that alloreactive T cells can effectively be depleted from allogeneic T cells by induction of AICD to prevent GvHD and might introduce a new strategy for the separation of GvH-reactive T cells and T cells mediating antiviral and possibly graft-versus-leukemia effects. PMID- 11929800 TI - High-dose therapy and autologous blood stem cell transplantation in POEMS syndrome. AB - We treated 5 patients with polyneuropathy, organomegaly, endocrinopathy, monoclonal gammopathy, and skin changes (POEMS) syndrome and multifocal bone lesions or diffuse bone marrow plasmacytic infiltration with high-dose therapy (HDT) and autologous blood stem cell transplantation. In all cases, the treatment produced remission of plasma cell proliferation associated with marked improvement in the patients' performance status, neurologic symptoms, and other manifestations of the syndrome. HDT with stem cell support should be investigated further as a therapeutic option in patients with POEMS syndrome and disseminated plasma cell dyscrasia. PMID- 11929799 TI - Randomized study of valacyclovir as prophylaxis against cytomegalovirus reactivation in recipients of allogeneic bone marrow transplants. AB - Oral valacyclovir for cytomegalovirus (CMV) prophylaxis in bone marrow transplantation (BMT) was investigated in a randomized, double-blind, acyclovir controlled, multicenter clinical trial in recipients of allogeneic BMT who were CMV seropositive (or donor positive) before transplantation and were aged 13 years or older. Patients were randomized before BMT. All initially received intravenous acyclovir (500 mg/m(2)) 3 times daily until day 28 after transplantation or after discharge, then oral valacyclovir (2 g) or acyclovir (800 mg) 4 times daily until week 18 after transplantation. Evidence of CMV infection, CMV disease, and death were documented for 22 weeks. Primary end points were time to CMV infection (detection of CMV in blood, broncho-alveolar lavage) or CMV disease and survival. Preemptive CMV therapy was permitted. Seven hundred twenty-seven patients were evaluable for efficacy. After the administration of intravenous acyclovir, valacyclovir was significantly more effective than oral acyclovir in reducing the incidence of CMV infection. CMV infection or disease developed in 102 (28%) valacyclovir patients, compared with 143 (40%) acyclovir patients (HR, 0.59; 95% CI, 0.46-0.76; P <.0001). Survival did not differ between treatments (76% and 75% in the valacyclovir and acyclovir groups, respectively). The safety of oral valacyclovir was similar to that of high-dose oral acyclovir. Valacyclovir was more effective than acyclovir in preventing CMV reactivation in BMT recipients and showed a similar safety profile. CMV disease incidence was low, and no differences were observed between oral valacyclovir and acyclovir. Survival was similar in each group. Valacyclovir prophylaxis provides a clinically valuable intervention but must be part of an overall strategy for CMV prevention in BMT. PMID- 11929801 TI - Loss of PU.1 expression is associated with defective immunoglobulin transcription in Hodgkin and Reed-Sternberg cells of classical Hodgkin disease. AB - Immunoglobulin transcription is impaired in Hodgkin and Reed-Sternberg (HRS) cells of classical Hodgkin disease (cHD). We recently demonstrated that defective immunoglobulin promoter transcription correlates with the down-regulation of the B-cell transcription factors Oct2 and BOB.1/OBF.1. These results prompted us to investigate whether immunoglobulin enhancer activity is also impaired in HRS cells and whether as yet unidentified factors could be necessary for immunoglobulin enhancer activity in HRS cells of cHD. Here we analyzed 30 cases of cHD for expression of the Ets family member PU.1 that is known to collaborate with multiple transcription factors and to regulate expression of immunoglobulin genes. We show that PU.1 is not expressed in primary and cultured HRS cells. Reintroduction of PU.1 and Oct2 in cultured HRS cells restored the activity of cotransduced immunoglobulin enhancer constructs. Our study identifies PU.1 deficiency as a recurrent defect in HRS cells that might contribute to their impairment of immunoglobulin transcription. PMID- 11929802 TI - Homozygous factor V splice site mutation associated with severe factor V deficiency. AB - We investigated a family whose proband has a severe bleeding disorder and factor V antigenic and functional levels of 8% and less than 1% of control values, respectively. Molecular analysis of the factor V gene revealed a novel homozygous mutation in the last nucleotide of exon 10. 1701G>T causes activation of a cryptic exonic splice site in exon 10, which encodes part of the factor V heavy chain (A2 domain). This leads to the deletion of 35 nucleotides and results in a frameshift with a premature stop codon at amino acid position 498. The G1701 and corresponding Gln509 are conserved in murine, bovine, and porcine factor V and in human factor VIII. Few factor V deficiency mutations have been identified as yet. Several are present in the heterozygous form in combination with factor V Leiden (Arg506Gln). This is the first reported homozygous splice site mutation in a patient with factor V deficiency. PMID- 11929803 TI - A novel mutation in the erythropoietin receptor gene is associated with familial erythrocytosis. AB - Primary familial erythrocytosis (familial polycythemia) is a rare myeloproliferative disorder with an autosomal dominant mode of inheritance. We studied a new kindred with autosomal dominantly inherited familial erythrocytosis. The molecular basis for the observed phenotype of isolated erythrocytosis is heterozygosity for a novel nonsense mutation affecting codon 399 in exon 8 of the erythropoietin receptor (EPOR) gene, encoding an EpoR peptide that is truncated by 110 amino acids at its C-terminus. The new EPOR gene mutation 5881G>T was found to segregate with isolated erythrocytosis in the affected family and this mutation represents the most extensive EpoR truncation reported to date, associated with familial erythrocytosis. Erythroid progenitors from an affected individual displayed Epo hypersensitivity in in vitro methylcellulose cultures, as indicated by more numerous erythroid burst-forming unit-derived colonies in low Epo concentrations compared to normal controls. Expression of mutant EpoR in interleukin 3-dependent hematopoietic cells was associated with Epo hyperresponsiveness compared to cells expressing wild-type EpoR. PMID- 11929804 TI - What do oncogenic mutations have to do with angiogenesis/vascular dependence of tumors? PMID- 11929805 TI - Differential mitotic responses to microtubule-stabilizing and -destabilizing drugs. AB - Although microtubule interacting agents inhibit spindle dynamics, thereby leading to a block in mitosis, we report that low concentrations of these drugs result in differential mitotic effects. Microtubule-stabilizing agents including Taxol, epothilone B, and discodermolide produce aneuploid populations of A549 cells in the absence of a mitotic block. Such aneuploid populations are diminished in an epothilone B-resistant cell line. In contrast, microtubule-destabilizing agents like colchicine, nocodazole, and vinblastine are unable to initiate aneuploidy. The aneuploid cells result from aberrant mitosis as multipolar spindles are induced by the stabilizing drugs, but not by destabilizing agents. The results suggest that the mechanism underlying aberrant mitosis may not be the same as that responsible for mitotic block, and that the former determines the sensitivity of cells to Taxol-like drugs. PMID- 11929806 TI - Biallelic inactivation of the thyroid hormone receptor beta1 gene in early stage breast cancer. AB - Loss of heterozygosity within the short arm of chromosome 3 is a common molecular event in several types of solid tumors. In breast cancer, 3p loss of heterozygosity occurs in invasive tumor cells as well as in morphologically normal terminal ductal lobular units adjacent to carcinoma in some cases [G. Deng et al., Science (Wash. DC), 274: 2057-2059, 1996.]. The most frequent region of allelic loss at 3p24.3 in morphologically normal terminal ductal lobular units encompasses the thyroid hormone receptor beta1 (TRbeta1) gene. Here we have observed a variable degree of TRbeta1 promoter hypermethylation in all 11 cases of primary breast cancer examined. Moreover, hypermethylation occurred at the same CpG sites in nonmalignant tissue peripheral to carcinoma in 4 of 11 cases. The lack of TRbeta1 nuclear staining, a likely result of biallelic gene inactivation, was observed in 25% (22 of 85) of primary tumors. This is a first demonstration of promoter hypermethylation and a concurrent reduction of TRbeta1 transcripts in breast cancer cell lines, although specific CpG sites targeted for gene silencing remain to be determined. Gene expression was restored by treatment with 5-aza-deoxycytidine in such cases. The observation of early, frequent, and multiple mechanisms of TRbeta1 inactivation suggests a potential role for this gene in the suppression of breast tumorigenesis. PMID- 11929807 TI - Endostatin regulates endothelial cell adhesion and cytoskeletal organization. AB - Endostatin, an endogenous angiogenesis inhibitor, attenuates endothelial cell migration through an unknown mechanism. We show that endostatin induced tyrosine phosphorylation of focal adhesion kinase and paxillin, and promoted formation of focal adhesions and actin stress fibers, similar to fibroblast growth factor-2 (FGF-2). In cells cotreated with endostatin and FGF-2, focal adhesions and actin stress fibers were decreased, indicating that endostatin disturbs cell-matrix adhesion. Reduced tyrosine phosphorylation and cytoplasmic relocalization of beta catenin in cells treated with FGF-2 and endostatin indicates that loosening of cell-cell adhesion is also disturbed by endostatin. These data provide a molecular basis both for the lack of effect of endostatin on the normal, quiescent vasculature, and its antagonistic effects on stimulated tumor vessels. PMID- 11929808 TI - Hormonal regulation of VEGF in orthotopic MCF7 human breast cancer. AB - Vascular endothelial growth factor (VEGF) mRNA and protein levels were higher in MCF7 breast cancer cells and orthotopic tumors after treatment with tamoxifen, as compared with treatment with estrogen. Accordingly, tumor vascular permeability, evaluated in vivo from contrast-enhanced magnetic resonance imaging, was elevated during treatment with tamoxifen. The results indicate that estrogen regulates angiogenesis in MCF7 tumors by maintaining VEGF at levels sufficient for the generation of functional microcapillaries and consequently facilitates tumor growth. During tamoxifen treatment, estrogen regulation is removed, and VEGF increases to levels that enhance markedly vascular permeability and reduce their perfusion function, leading to inhibition of tumor progression. PMID- 11929809 TI - High frequency loss of heterozygosity in von Hippel-Lindau (VHL)-associated and sporadic pancreatic islet cell tumors: evidence for a stepwise mechanism for malignant conversion in VHL tumorigenesis. AB - Germ-line mutation of the von Hippel-Lindau (VHL) gene predisposes to the development of multifocal, benign lesions, including retinal and central nervous system hemangioblastomas, pheochromocytomas, and renal and pancreatic cysts. Progression to malignancy in VHL disease is associated primarily with the development of renal cell carcinoma (RCC) and pancreatic islet cell tumors (PICT). Although many reports have documented the multiple functions of the VHL protein, few have investigated the intriguing question related to the tissue specificity of malignant conversion in VHL disease, a problem not easily explained by strict genotype-phenotype correlations. We investigated a novel VHL kindred with a preponderance of PICTs to determine whether loss of additional genetic loci associated with the sporadic forms of RCC and PICTs might play a role in malignant conversion in this disease. We report the high frequency loss of heterozygosity (LOH) of genetic loci distinct from and mapping proximal to VHL within human chromosome 3p in the VHL kindred under study. Furthermore, chromosome 3p LOH occurs subsequent to VHL mutation and cyst formation, and correlates with malignant progression in VHL-associated PICTs. High frequency LOH was also observed in sporadic PICTs in regions of 3p associated with LOH in sporadic clear cell RCC as well as homozygous deletion in lung cancer. A stepwise model for malignant conversion in VHL disease is herein proposed. PMID- 11929810 TI - Lack of mutation induction with exposure to 1.5 GHz electromagnetic near fields used for cellular phones in brains of Big Blue mice. AB - The possible mutagenic potential of exposure to 1.5 GHz electromagnetic near field (EMF) was investigated using brain tissues of Big Blue mice (BBM). Male BBM were locally exposed to EMF in the head region at 2.0, 0.67, and 0 W/kg specific absorption rate for 90 min/day, 5 days/week, for 4 weeks. No gliosis or degenerative lesions were histopathologically noted in brain tissues, and no obvious differences in Ki-67 labeling and apoptotic indices of glial cells were evident among the groups. There was no significant variation in the frequency of independent mutations of the lacI transgene in the brains. G:C to A:T transitions at CpG sites constituted the most prevalent mutations in all groups and at all time points. Deletion mutations were slightly increased in both the high and low EMF exposure groups as compared with the sham-exposed group, but the differences were not statistically significant. These findings suggest that exposure to 1.5 GHz EMF is not mutagenic to mouse brain cells and does not create any increased hazard with regard to brain tumor development. PMID- 11929811 TI - Low mutation incidence in polymorphic noncoding short mononucleotide repeats in gastrointestinal cancer of the microsatellite mutator phenotype pathway. AB - Frameshifts in short mononucleotide tracts (SMT) in genes, such as TGFbetaRII and BAX, are common in gastrointestinal tumors of the microsatellite mutator phenotype (MMP). The significance of less common mutations has been recently challenged because frequencies as high as 50% were reported in some noncoding SMTs in MMP colon cancer cell lines (L. Zhang, et al., Cancer Res., 61: 3801 3805, 2001). We did not confirm these findings after examining >50 MMP gastrointestinal cancers for mutations in eight SMT loci with the highest reported frequencies. In three of these loci, no clonal mutations were detected, and they were infrequent (2.9-6.7%) in the other five. Length polymorphisms are frequent (25.7-43.9%) in one-half of these SMTs, suggesting an explanation for the discrepancy. Because of the peculiar features of MMP tumors, low prevalence of mutations in cancer genes may not be a disqualifying criterion for their functionality. PMID- 11929812 TI - Metabolic markers of breast cancer: enhanced choline metabolism and reduced choline-ether-phospholipid synthesis. AB - Specific genetic alterations during malignant transformation may induce the synthesis and breakdown of choline phospholipids, mediating transduction of mitogenic signals. The high level of water-soluble choline metabolites in cancerous breast tumors, relative to benign lesions and normal breast tissue, has been used as a diagnostic marker of malignancy. To unravel the biochemical pathways underlying this phenomenon, we used tracer kinetics and 13C and 31P magnetic resonance spectroscopy to compare choline transport, routing, and metabolism to phospholipids in primary cultures of human mammary epithelial cells and in MCF7 human breast cancer cells. The rate of choline transport under physiological choline concentrations was 2-fold higher in the cancer cells. The phosphorylation of choline to phosphocholine and oxidation of choline to betaine yielded 10-fold higher levels of these metabolites in the cancer cells. However, additional incorporation of choline to phosphatidylcholine was similar in both cell types. Thus, enhanced choline transport and augmented synthesis of phosphocholine and betaine are dominant pathways responsible for the elevated presence of choline metabolites in cancerous breast tumors. Uniquely, reduced levels and synthesis of a choline-ether-phospholipid may also serve as a metabolic marker of breast cancer. PMID- 11929813 TI - Lack of tumorigenesis in the mouse liver after adenovirus-mediated expression of a dominant stable mutant of beta-catenin. AB - Mutations in the glycogen synthase kinase 3beta (GSK3beta) phosphorylation sites of the beta-catenin gene exon 3 are found in 20-30% of human primary hepatocellular carcinoma (HCC), whereas mutations in the APC or AXIN genes are found in other HCC populations. These data strongly suggest that the Wnt signaling pathway is involved in hepatocarcinogenesis. To determine the role of beta-catenin in intestinal tumorigenesis, we earlier constructed a mutant mouse strain Catnb(lox(ex3)), in which exon 3 of the beta-catenin gene was sandwiched by loxP sequences. By genetic crosses of these mice with the Fabpl-cre transgenic mice that express the cre gene controlled by the fatty acid binding protein gene promoter, we introduced the beta-catenin stabilizing mutation into the small intestine and liver. Although numerous polyps were formed in the small intestine, we did not find any neoplastic (i.e., dysplastic) foci in the liver, and the mice died in 5 weeks after birth because of acute liver damage accompanying mitochondrial swelling. When a recombinant adenovirus that expresses the cre gene from a human cytomegalovirus early gene promoter was constructed and inoculated at a high multiplicity (10(9) plaque-forming units/mouse), the Catnb(lox(ex3)) mice showed marked hepatomegaly, with similar mitochondrial swelling in the hepatocytes, and died within 3 weeks after infection. On the other hand, when inoculated at lower multiplicities of infection (10(7) and 10(8) plaque-forming units/mouse, respectively), the Catnb(lox(ex3)) mice survived >6 months without any neoplastic foci in the liver, although the nuclear localization of beta catenin was found in some hepatocytes even after 6 months. These results suggest that, in contrast to intestinal polyposis, the Wnt pathway activation by stabilized beta-catenin is not sufficient for hepatocarcinogenesis, but additional mutations or epigenetic changes may be required. PMID- 11929814 TI - Characterization of benzo(a)pyrene-trans-7,8-dihydrodiol glucuronidation by human tissue microsomes and overexpressed UDP-glucuronosyltransferase enzymes. AB - UDP-glucuronosyltransferase (UGT)-mediated glucuronidation of benzo(a)pyrene trans-7,8-dihydrodiol (BPD), precursor to the potent mutagen benzo(a)pyrene-7,8 dihydrodiol-9,10-epoxide, may be an important pathway in the detoxification of benzo(a)pyrene. To better characterize this pathway in humans, high-pressure liquid chromatography (HPLC) was used to detect glucuronide conjugates of BPD formed in vitro. Three peaks were detected by HPLC after incubation of racemic BPD with human liver microsomes; these were identified as monoglucuronides by liquid chromatography-mass spectrometry analysis. Proton nuclear magnetic resonance spectroscopy of isolated fractions, combined with HPLC analysis of the glucuronide products from human liver microsomal incubations with purified benzo(a)pyrene-trans-7S,8S-dihydrodiol [(+)-BPD] and benzo(a)pyrene-trans-7R,8R dihydrodiol [(-)-BPD] forms of BPD, indicated that peak 1 contained the 7 glucuronide of 7S,8S-BPD (BPD-7S-Gluc), peak 2 was a mixture of the 7-glucuronide of 7R,8R-BPD (BPD-7R-Gluc) and the 8-glucuronide of 7S,8S-BPD (BPD-8S-Gluc), and peak 3 contained the 8-glucuronide of 7R, 8R-BPD (BPD-8R-Gluc). In liver microsomes, peak 1 (BPD-7S-Gluc) was the largest peak observed, whereas in microsomes from aerodigestive tract tissues, peak 2 (both BPD-7R-Gluc and BPD-8S Gluc) was the largest HPLC peak observed. The liver enzymes UGT1A1 and UGT2B7 formed BPD-7S-Gluc as the major diastereomer, whereas UGT1A8 and UGT1A10, extrahepatic enzymes present in the aerodigestive tract, preferentially formed both BPD-7R-Gluc and BPD-8S-Gluc. In addition, both UGT1A9 and UGT1A7 preferentially formed BPD-7R-Gluc. No detectable glucuronidating activity against BPD was observed by UGT1A3, UGT1A4, UGT1A6, UGT2B4, UGT2B15, or UGT2B17. The affinity of individual UGT enzymes as determined by K(m) analysis was UGT1A10 > UGT1A9 > UGT1A1 > UGT1A7 for (-)-BPD and UGT1A10 > UGT1A9 > UGT2B7 approximately UGT1A1 > UGT1A7 for (+)-BPD. These results suggest that several UGTs may play an important role in the overall glucuronidation of BPD in humans, with UGT1A1, UGT1A7, UGT1A9, UGT1A10 and potentially UGT1A8 playing an important role in the glucuronidation of the procarcinogenic (-)-BPD enantiomer, and that the stereospecific activity exhibited by different UGTs against BPD is consistent with tissue-specific patterns of BPD glucuronide diastereomer formation and UGT expression. PMID- 11929815 TI - Prevalence and spectrum of p53 mutations associated with smoking in breast cancer. AB - To explore the role of smoking in breast cancer, we undertook a population-based study to evaluate the prevalence and spectrum of p53 mutations in the breast tumors of smokers and nonsmokers. We evaluated 456 archival invasive breast tumors for mutations in exons 4-8 of the p53 gene, using single-strand conformational polymorphism analysis and manual sequencing. Statistical analyses were performed to determine the association of p53 mutations with clinical and smoking characteristics. Of 108 mutations identified, 77 (71%) were point mutations and 31 (29%) were deletions or insertions. A higher prevalence of p53 mutations was found in the breast tumors of current smokers (36.5%; P = 0.02) compared with never smokers (23.6%), whereas fewer mutations were found in former smokers (16.2%; P = 0.09). After adjustment for age, race, menopausal status, clinical stage, tumor size, and family history of breast cancer, current smokers were significantly more likely to harbor any p53 mutation [odds ratio (OR), 2.11; 95% confidence interval (CI), 1.17-3.78], p53 transversions (OR, 3.37; 95% CI, 1.03-11.06), and G:C-->T:A transversions (OR, 10.53; 95% CI, 1.77-62.55) compared with never smokers. Stage at diagnosis did not account for the increase in p53 mutation-positive breast cancer among current smokers. Former smokers were also more likely than never smokers to harbor G:C-->T:A transversions (OR, 2.43; 95% CI, 0.37-15.73), although this association was not statistically significant. Among former smokers, the prevalence of p53 mutations varied with time since quitting: former smokers who quit smoking for longer than 1 year had a lower prevalence of p53 mutations (10.5% for 1-5 years and 12.9% for >5 years) than those who had stopped smoking within the year of their cancer diagnosis (26.3%). Our results indicate that cigarette smoking appears to modify the prevalence and spectrum of p53 mutations in breast tumors. Moreover, the difference in mutational spectra observed between smokers and nonsmokers is suggestive of the genotoxic effects of smoking in breast tissue. PMID- 11929816 TI - Blockade of vascular endothelial growth factor receptor and epidermal growth factor receptor signaling for therapy of metastatic human pancreatic cancer. AB - We determined whether concurrent blockage of vascular endothelial growth factor (VEGF) receptor and epidermal growth factor (EGF) receptor signaling by two novel tyrosine kinase inhibitors, PTK 787 and PKI 166, respectively, can inhibit angiogenesis and, hence, the growth and metastasis of human pancreatic carcinoma in nude mice. Highly metastatic human pancreatic carcinoma L3.6pl cells were injected into the pancreas of nude mice. Seven days later, groups of mice began receiving oral doses of PTK 787 and PKI 166 three times weekly. Some groups of mice also received i.p. injections of gemcitabine twice a week. The mice were necropsied when the control mice became moribund. Treatment with PTK 787 and PKI 166, with gemcitabine alone, or with the combination of PTK 787, PKI 166, and gemcitabine produced 69, 50, and 97% reduction in the volume of pancreatic tumors, respectively. Administration of protein tyrosine kinase inhibitors and gemcitabine also significantly decreased the incidence of lymph node and liver metastasis. The therapeutic efficacy directly correlated with a decrease in circulating proangiogenic molecules (VEGF, interleukin-8), a decrease in microvessel density, a decrease in proliferating cell nuclear antigen staining, and an increase in apoptosis of tumor cells and endothelial cells. Therapies produced by combining gemcitabine with either PKI 166 or PTK 787 were similar to those produced by combining gemcitabine with both PKI 166 and PTK 787. These results suggest that blockade of either epidermal growth factor receptor or VEGF receptor signaling combined with chemotherapy provides an effective approach to the therapy of pancreatic cancer. PMID- 11929817 TI - Systemic inhibition of tumor growth and angiogenesis by thrombospondin-2 using cell-based antiangiogenic gene therapy. AB - Recent studies indicate that continuous administration improves the antitumoral efficacy of angiogenesis inhibitors, as compared with intermittent dosing, suggesting a potential role of gene therapy in antiangiogenic tumor therapy. We established a tissue-engineered implant system for the continuous in vivo production of thrombospondin-2 (TSP-2), a potent endogenous inhibitor of tumor growth and angiogenesis. Fibroblasts were retrovirally transduced to overexpress TSP-2 and were seeded onto biodegradable polymer scaffolds. After transplantation into the peritoneal cavity of nude mice, bioimplants maintained high levels of TSP-2 secretion over extended time periods, resulting in increased levels of circulating TSP-2. Bioimplant-generated TSP-2 potently inhibited tumor growth and angiogenesis of human squamous cell carcinomas, malignant melanomas, and Lewis lung carcinomas that were implanted at a distant site. These results provide the first proof-of-principle for the feasibility and therapeutic efficiency of systemic, cell-based antiangiogenic gene therapy using biodegradable polymer grafts for the treatment of cancer. PMID- 11929819 TI - Soluble FLT-1 expression suppresses carcinomatous ascites in nude mice bearing ovarian cancer. AB - Vascular endothelial growth factor (VEGF), a bifunctional protein enhancing vascular permeability and stimulating endothelial growth, is thought to be responsible for fluid accumulation and angiogenesis in ascites tumors. To investigate the effects of stable expression of the soluble form of Flt-1 VEGF receptor (sFlt-1), a known endogenous inhibitor of VEGF, on the malignant ascites tumors, we cotransduced RMG-1 human ovarian cancer cells with adeno-associated virus vectors carrying the sFlt-1 cDNA and Neo gene or Neo gene alone and isolated both the sFlt-1-expressing clone and the Neo-expressing clone. In vitro growth characteristics were essentially the same. As expected, conditioned medium collected from the sFlt-1-expressing cells significantly inhibited the human umbilical vein endothelial cell proliferation in the presence of recombinant VEGF. Expression of sFlt-1 significantly suppressed RMG-1 cell-induced angiogenesis in vivo in the mouse dorsal air sac assay model. We then inoculated sFlt-1- or Neo alone-expressing cells i.p. into female BALB/c nude mice. The average volume of ascites fluid, number of leaked RBCs, and number of cancer cells were significantly lower in mice injected with sFlt-1-expressing cells than in the controls. Survival time was significantly prolonged in mice injected with sFlt-1-expressing cells. These results suggest that inhibition of VEGF activity by sFlt-1 expression may provide a means to control carcinomatous ascites and angiogenesis of malignant ascites tumors. PMID- 11929818 TI - Antitumor effect of TAT-oxygen-dependent degradation-caspase-3 fusion protein specifically stabilized and activated in hypoxic tumor cells. AB - Human solid tumors contain hypoxic regions that have considerably lower oxygen tension than normal tissues. These impart resistance to radiotherapy and anticancer chemotherapy, as well as predisposing to increased tumor metastases. To develop a potentially therapeutic protein drug highly specific for solid tumors, we constructed fusion proteins selectively stabilized in hypoxic tumor cells. A model fusion protein, oxygen-dependent degradation (ODD)-beta galactosidase (beta-Gal), composed of a part of the ODD domain of hypoxia inducible factor-1alpha fused to beta-Gal, showed increased stability in cultured cells under a hypoxia-mimic condition. When ODD-beta-Gal was further fused to the HIV-TAT protein transduction domain (TAT(47-57)) and i.p. injected to a tumor bearing mouse, the biologically active fusion protein was specifically stabilized in solid tumors but was hardly detected in the normal tissue. Furthermore, when wild-type (WT) caspase-3 (Casp3(WT)) or its catalytically inactive mutant was fused to TAT-ODD and i.p. injected to a tumor-bearing mouse, the size of tumors was reduced by the administration of TAT-ODD-Casp3(WT) but not by TAT-ODD-mutant Casp3. TAT-ODD-Casp3(WT) did not cause any obvious side effects on tumor-bearing mice, suggesting specific stabilization and activation of the fusion protein in the hypoxic tumor cells. These results suggest that the combination of protein therapy using a cytotoxic TAT-ODD fusion protein with radiotherapy and chemotherapy may provide a new strategy for annihilating solid tumors. PMID- 11929820 TI - Tumor genotype-specific growth inhibition in vivo by antisense oligonucleotides against a polymorphic site of the large subunit of human RNA polymerase II. AB - Loss of heterozygosity (LOH) reduces genes to hemizygosity in cancer cells and presents an absolute difference between normal and cancer cells. The regions of LOH are usually much larger than the tumor suppressor gene, which is lost, and are expected to contain genes that are essential for cell survival. Single nucleotide polymorphisms (SNPs) are the most common type of genetic variation in man, often giving rise to two or more allelic forms of most genes. SNPs of essential genes that are frequently affected by LOH can be used as a target for a novel therapy against cancer cells with LOH. The SNPs can be targeted by antisense oligonucleotides (ODNs) that will discriminate between two alleles. We have designed allele-specific phosphorothioate ODNs against the gene of the large subunit of RNA polymerase II (POLR2A), a gene located in close proximity to the tumor suppressor gene p53, which frequently shows LOH in cancer cells. This report shows that phosphorothioate antisense ODNs directed against POLR2A can inhibit tumor growth in vivo as efficiently as a well-described antitumor antisense ODN directed against Ha-ras. In addition, we show that a single bp mismatch can be sufficient to obtain allele-specific inhibition of tumor growth, demonstrating that the effects observed are true antisense effects. PMID- 11929821 TI - Celecoxib exhibits the greatest potency amongst cyclooxygenase (COX) inhibitors for growth inhibition of COX-2-negative hematopoietic and epithelial cell lines. AB - Cyclooxygenase-2 (COX-2) is an important cellular target for both therapy and/or prevention of inflammatory disorders and cancer. The advent of selective COX-2 inhibitors now allows a more precise and safer treatment approach. The screening of an array of cancer cell lines for growth inhibitory effects of COX-2-selective and -nonselective inhibitors, including celecoxib (Celebrex) and rofecoxib (Vioxx), produced two unanticipated findings. Firstly, the antiproliferative effects of celecoxib were noted to be of very similar magnitude for both hematopoietic and epithelial cancer cell lines. Most hematopoietic cell lines had no detectable COX-2 expression by reverse transcription-PCR, and none expressed COX-2 protein. In addition, COX-2-negative epithelial lines were found to have IC50s for celecoxib that were very similar to their COX-2+ counterparts. Thus, important antiproliferative effects were observed that were independent of both the cell lineage and COX-2 status. Secondly, it was also observed that COX-2 inhibitor drugs, celecoxib and rofecoxib, with similar COX-2-selectivity and clinical efficacy for inflammatory indications, differed significantly in their in vitro antiproliferative effects on cancer cell lines. IC50s of 35-65 microM were observed for celecoxib across this entire panel of cell lines. Finally, no difference in the mode or degree of cytotoxicity was apparent between cell lines, because similar levels of apoptosis were observed in COX-2+ and -negative cell lines after treatment with celecoxib, with correspondingly lower levels after rofecoxib treatment. These data are important in that they provide the first direct comparison of epithelial and hematopoietic cancer cell lines, as well as a direct comparison of the in vitro anticancer effects of the two clinically available COX-2 inhibitors. PMID- 11929822 TI - Combined anti-fetal liver kinase 1 monoclonal antibody and continuous low-dose doxorubicin inhibits angiogenesis and growth of human soft tissue sarcoma xenografts by induction of endothelial cell apoptosis. AB - Vascular endothelial growth factor (VEGF) and VEGF receptor 2 [fetal liver kinase 1 (Flk-1)/kinase insert domain-containing receptor] have been shown to play a major role in tumor angiogenesis. In this study, we investigated whether anti-Flk 1 monoclonal antibody DC101 could therapeutically inhibit growth and angiogenesis of human soft tissue sarcoma, and we explored its capacity to enhance the tumoricidal effects of doxorubicin. Treatment of well-established leiomyosarcoma SKLMS-1 and rhabdomyosarcoma RD xenografts in severe combined immunodeficient mice with DC101 resulted in significant antitumor activity. In a parallel study, we compared tumor inhibition with continuous low-dose "antiangiogenic" schedule versus once-every-2-weeks high-dose standard schedule of doxorubicin. We found that continuous low-dose treatment inhibited the tumor growth of RD xenografts about 46.5% of that with standard-schedule treatment, but that continuous low dose treatment did not inhibit the tumor growth of SKLMS-1 xenografts. Notably, combined DC101 and continuous low-dose doxorubicin resulted in more effective growth inhibition of SKLMS-1 and RD xenografts than has been observed with any agent alone in a long-term s.c. tumor xenograft model. The combination therapy was associated with no additional toxicity to the host animal compared with low dose doxorubicin alone. Histological examination of xenografts showed significantly reduced microvessel counts in the tumors given combined therapy compared with the tumors given either agent alone. These results are consistent with an enhanced inhibition of angiogenesis in vivo by combined DC101 and doxorubicin using Matrigel plug assay. Additionally, DC101 plus doxorubicin directly exerted enhanced inhibitory effects on endothelial cell migration, proliferation, and tube-like formation in vitro. Furthermore, the combination induced an enhanced apoptosis of endothelial cells that was associated with an increase of capase-3 activity. Thus, the inhibition of angiogenesis and induction of endothelial cell apoptosis are likely important mechanisms for the antitumor activity of combined DC101 and doxorubicin. Collectively, our data suggested that anti-VEGF receptor 2 in combination with continuous low-dose doxorubicin may provide a new therapeutic approach for human soft tissue sarcoma in the clinic. PMID- 11929823 TI - Synergistic enhancement of antitumor immunity with adoptively transferred tumor specific CD4+ and CD8+ T cells and intratumoral lymphotactin transgene expression. AB - The lack of efficient T-cell infiltration of tumors is a major obstacle to successful adoptive T-cell therapy. We have shown that transplanted SP2/0 myeloma tumors that have been engineered to express lymphotactin (Lptn) invariably regress under the influence of infiltrating XCR1+T cells and neutrophils. Herein, we characterize these T cells and investigate their therapeutic efficacy, either alone or with Lptn gene therapy. After stimulation with SP2/0 cells, these T cells were CD25+FasL+L-selectin-, expressed XCR-1, and were chemoattracted by Lptn in vitro. They comprised 66% CD4+ Th1 and 33% CD8+ Tc1 cells, both of which expressed significant amounts of IFN-gamma, perforin, and tumor necrosis factor alpha, but not interleukin-4. The CD4+ Th1 and CD8+ Tc1 cells, which were inhibited and stimulated, respectively, for proliferation with Lptn signaling, displayed 38 and 84% specific killing, respectively, for Ia(d)/H-2K(d)-expressing SP2/0 tumor cells (E:T ratio, 100). In vivo, combined intratumoral Lptn gene transfer and adoptive immunotherapy with these CD4+ and CD8+ T cells eradicated well-established SP2/0 tumors in six of eight mice, and dramatically slowed tumor growth in the other two mice. Cell tracking using labeled T cells confirmed that these cells infiltrated better into the Lptn-expressing tumors than non-Lptn expressing ones. Control or Lptn adenoviral treatments by themselves did not alter the lethal outcome for tumor-bearing mice, nor did T-cell therapy by itself, although the latter two treatments did slow its time frame. Combined Lptn gene transfer and adoptive CD4+ or CD8+ cell transfers were not nearly as efficacious as the combined Lptn gene and unfractionated T-cell transfers. Taken together, our data provide solid evidence of a potent synergy between adoptive CD4+ and CD8+ T-cell therapy and Lptn gene transfer into tumor tissues, which culminated in the eradication of well-established tumor masses. PMID- 11929824 TI - CD40-expressing carcinoma cells induce down-regulation of CD40 ligand (CD154) and impair T-cell functions. AB - The interaction of CD40 expressed by immunocompetent cells with its ligand CD154 on the surface of T-helper cells plays a crucial role in the immune response. Recently, the presence of CD40 was also demonstrated on a variety of carcinomas. Whereas the critical relevance of CD40 in cytotoxic T-cell priming via dendritic cells is already established, the biological role of CD40/CD154 interactions in nonhematopoetic cells is still unclear. In the present study we demonstrate that CD154 expression density is down-regulated on activated T cells on interaction with CD40+ tumor cells. Naive T cells cocultured with CD40+carcinoma showed impaired functionality as indicated by a reduced frequency of IFN-gamma secreting cells, reduced interleukin 2 secretion, impaired proliferation, and a lack of CD154 re-expression on restimulation. In distinction, T-cell effector lysing capacity was not impaired by CD40-expressing tumor cell targets. The present results suggest that in marked contrast to antigen-presenting cells, CD40 expression on carcinoma cells suppresses T-cell activation. Our findings support the statement that CD40 functions are context dependent and imply a new function for CD40 expressed on nonantigen-presenting cells. PMID- 11929825 TI - Toward synthetic combinatorial peptide libraries in positional scanning format (PS-SCL)-based identification of CD8+ Tumor-reactive T-Cell Ligands: a comparative analysis of PS-SCL recognition by a single tumor-reactive CD8+ cytolytic T-lymphocyte clone. AB - The use of synthetic combinatorial peptide libraries in positional scanning format (PS-SCL) has emerged recently as an alternative approach for the identification of peptides recognized by T lymphocytes. The choice of both the PS SCL used for screening experiments and the method used for data analysis are crucial for implementing this approach. With this aim, we tested the recognition of different PS-SCL by a tyrosinase 368-376-specific CTL clone and analyzed the data obtained with a recently developed biometric data analysis based on a model of independent and additive contribution of individual amino acids to peptide antigen recognition. Mixtures defined with amino acids present at the corresponding positions in the native sequence were among the most active for all of the libraries. Somewhat surprisingly, a higher number of native amino acids were identifiable by using amidated COOH-terminal rather than free COOH-terminal PS-SCL. Also, our data clearly indicate that when using PS-SCL longer than optimal, frame shifts occur frequently and should be taken into account. Biometric analysis of the data obtained with the amidated COOH-terminal nonapeptide library allowed the identification of the native ligand as the sequence with the highest score in a public human protein database. However, the adequacy of the PS-SCL data for the identification for the peptide ligand varied depending on the PS-SCL used. Altogether these results provide insight into the potential of PS-SCL for the identification of CTL-defined tumor-derived antigenic sequences and may significantly implement our ability to interpret the results of these analyses. PMID- 11929826 TI - Hepatocyte growth factor induces Wnt-independent nuclear translocation of beta catenin after Met-beta-catenin dissociation in hepatocytes. AB - Hepatocyte growth factor (HGF) and Wnt signaling pathways have been shown to be important in embryogenesis and carcinogenesis. The aim of this study was to elucidate the mechanism of functional similarities observed in the two pathways. We used normal rat liver, primary hepatocyte cultures and a dominant-negative Met expression system to study the effect of HGF on Wnt pathway components. We demonstrate novel association of beta-catenin and Met, a tyrosine kinase receptor of HGF, at the inner surface of the hepatocyte membrane. HGF induces dose dependent nuclear translocation of beta-catenin in primary hepatocyte cultures that is Wnt independent. The source of beta-catenin for translocation in hepatocytes is the Met-beta-catenin complex, which appears to be independent of the E-cadherin-beta-catenin complex. To test the functionality of this association, we used a dominant-negative Met expression system that expresses only the extracellular and transmembrane regions of the beta-subunit of Met. A loss of Met-beta-catenin association resulted in abrogation of nuclear translocation of beta-catenin upon HGF stimulation. This event is tyrosine phosphorylation dependent, and the association of Met and beta-catenin is crucial for this event. We conclude that the HGF causes similar redistribution of beta catenin as Wnt-1 in the hepatocytes and that this effect is attributable to subcellular association of Met and beta-catenin. The intracellular kinase domain of Met is essential for tyrosine phosphorylation and nuclear translocation of beta-catenin. Part of the multifunctionality of HGF might be attributable to nuclear beta-catenin and the resulting target gene expression. PMID- 11929827 TI - Epigenetic inactivation of 14-3-3 sigma in oral carcinoma: association with p16(INK4a) silencing and human papillomavirus negativity. AB - In vitro studies have identified 14-3-3sigma as a regulator of senescence in human keratinocytes. To assess its contribution to squamous neoplasia, we have analyzed genetic and epigenetic changes in this gene in squamous cell carcinomas (SCCs) and dysplastic lesions of the oral cavity. No mutations were detected in the coding sequence of 14-3-3sigma in 20 oral carcinomas, and there was loss of heterozygosity in only 7 of 40 informative cases. In contrast to the absence of genetic change, aberrant methylation within 14-3-3sigma was detected in 32 of 92 squamous cell carcinomas and in 3 of 6 oral dysplasias and was associated with reduced or absent expression at both mRNA and protein levels. Methylation was not detected in matched, normal epithelial tissue controls. Carcinomas in which 14-3 3sigma was methylated were significantly more likely to lack DNA sequences from human papillomavirus and to have coincident methylation of p16(INK4a) than cases that expressed 14-3-3sigma. Methylation was detected in SCC, both wild-type and mutant for p53, but was more commonly detected in cancers with wild-type p53. These results implicate coincident epigenetic abrogation of function in both sigma and p16(INK4a) in a subset of SCCs of the oral cavity. PMID- 11929829 TI - Comparative gene expression profile analysis of neurofibromatosis 1-associated and sporadic pilocytic astrocytomas. AB - Pilocytic astrocytomas (PAs) are WHO grade I brain tumors that do not typically progress to more malignant grades of astrocytoma. Whereas there have been significant advances in the molecular genetics of high-grade astrocytomas, relatively little is known about the genetic changes associated with PA formation. In an effort to better characterize these low-grade neoplasms, we compared the gene expression profiles of six sporadic and two neurofibromatosis 1 associated PAs with other tissues and cell lines of both astrocytic and oligodendroglial origin. Hierarchical cluster analysis of gene expression data clearly delineated PAs from low-grade oligodendrogliomas and normal white matter. The two NF1-associated tumors and one of the sporadic PAs displayed expression profiles that were more closely related to those of cultured normal human fetal astrocytes. However, PAs also expressed individual genes typically associated with oligodendroglial lineage (e.g., proteolipid protein and PMP-22). The expression patterns of specific genes (e.g., ApoD) were unique to PA tumors, whereas genetic changes characteristic of high-grade astrocytomas were not encountered. Differential expression of two transcripts, neural cellular adhesion molecule and connexin-43, was confirmed at the protein level, suggesting that these cell adhesion molecules might be particularly important in the molecular pathogenesis of these tumors. We conclude that PAs are genetically unique gliomas with gene expression profiles that resemble those of fetal astrocytes and, to a lesser extent, oligodendroglial precursors. PMID- 11929828 TI - Differential effects of p21(WAF1/CIP1) deficiency on MMTV-ras and MMTV-myc mammary tumor properties. AB - p21(WAF1/CIP1) (p21) functions as a cyclin-dependent kinase (CDK) inhibitor and is a key mediator of p53-dependent growth arrest. However, its role in cell cycle regulation is complex, because it also appears to promote CDK activity in certain experimental contexts. Its potential role in tumor suppression was evaluated in MMTV-ras and MMTV-myc transgenic mice that were interbred to p21(WAF1/CIP1) knockout mice (p21-/-). p21 deficiency had differential effects on tumor incidence and age of onset, proliferation, and apoptosis in the presence of these two oncogenes. Tumors arising in MMTV-ras/p21-/- mice displayed higher S-phase fractions and correspondingly increased cyclin D1 and E/CDK activity than MMTV ras tumors. In contrast, MMTV-myc/p21-/- tumors had lower S-phase fractions and levels of cyclin D1 and E/CDK activity than MMTV-myc tumors. In both tumor types, changes in cyclin D1 and E/CDK activity were paralleled by changes in the corresponding cyclin protein levels. Tumor cell apoptosis was also differentially influenced by p21 deficiency in the two models. MMTV-ras/p21-/- tumors exhibited a significant increase in spontaneous apoptosis as compared with MMTV-ras tumors, whereas p21 deficiency had minimal effect on apoptosis in MMTV-myc tumors. These results indicate that the effects of p21 expression on cellular proliferation are differentially affected by the expression of different oncogenes, and that p21 may play a role in promoting either growth arrest or proliferation, depending on the specific cellular context. PMID- 11929830 TI - Susceptibility of Msh2-deficient mice to inflammation-associated colorectal tumors. AB - Patients with longstanding extensive ulcerative colitis have an increased risk of developing colorectal cancer (CRC). There are significant differences in the early pathogenesis of colitis-associated tumors compared with common CRC, whereas the frequency, degree, and significance of microsatellite instability (MSI) as a marker of mismatch repair deficiency in colitis tumors remain unclear. Here we describe the application of the DSS model of chronic colitis to mice with a defect in the Msh2 mismatch repair gene to discern these early events. These mice do not develop CRC spontaneously without an external trigger. The aim of this study was to determine the effect of the Msh2 defect on the frequency and grade of colitis-associated colorectal dysplasia and adenocarcinoma in Msh2-/-, Msh2+/ , and wild-type (Msh2+/+) mice and on the MSI status of the tumors. We show that in mice with chronic colitis, 60% of the Msh2-/- and 29% of the wild-type mice developed high-grade dysplasia or adenocarcinoma, but heterozygosity for the Msh2 defect did not increase tumor susceptibility over wild-type genotype. The largest difference between genotypes was in the frequency of high-grade dysplasia, with 46.7, 8, and 12.5% in Msh2-/-, Msh2+/-, and Msh2+/+ mice, respectively. The Msh2 /- mice developed MSI-high tumors, whereas the majority of the Msh2+/- and wild type tumors had no MSI. In the Msh2-/- mice, MSI appeared early in non-neoplastic colon tissue, presumably as a result of markedly increased epithelial cell proliferation associated with inflammation. These observations suggest that a homozygous mismatch repair defect predisposes to tumors triggered by chronic inflammation but is not the only factor involved because tumors also developed in the wild-type mice. This model of colitis offers opportunities to characterize the different molecular pathways of carcinogenesis operating in chronic colitis. PMID- 11929831 TI - The melanocyte-specific isoform of the microphthalmia transcription factor affects the phenotype of human melanoma. AB - The microphthalmia transcription factor MITF plays a pivotal role in the development and differentiation of melanocytes. The purpose of this work was to investigate the expression and function of the melanocyte-specific isoform MITF-M in human melanoma. We found that MITF-M is repressed in 8 of 14 established melanoma cell lines tested. Transfection of MITF-M into a melanoma cell line (518A2) lacking the M-isoform and into a permanent cell line established from normal melanocytes (NMel-II) resulted in slower tumor growth in a severe combined immunodeficient-mouse xenotransplantation model. The growth difference between vector control-transfected tumors derived from the NMel-II cell line (mean tumor weight +/- SD, 3.2 g +/- 1.13) and MITF-M (+) transfectants (mean tumor weight +/ SD, 1.1 g +/- 0.49) was significant (P = 0.018). The mean tumor weight of control-transfected 518A2 tumors was 0.99 g +/- 0.22 and of MITF-M (+) transfectants, 0.69 g +/- 0.32. The difference in growth between 518A2 controls and the MITF-M (+) transfectants was clear, however it did not reach statistical significance (P = 0.08). In addition to the growth-inhibitory effects, MITF-M expression led to a change in the histopathological appearance of tumors from epitheloid toward a spindle-cell type in vivo. These results indicate a role for the MITF-M isoform in the in vivo growth control and the phenotype of human melanoma. In conclusion, MITF-M may qualify as a marker capable of identifying subgroups of melanoma patients with different tumor biology and prognosis. PMID- 11929832 TI - Limited proliferation and telomere dysfunction following telomerase inhibition in immortal murine fibroblasts. AB - Telomerase is a ribonucleoprotein enzyme that functions to maintain telomeres, the terminal DNA that protects chromosomal integrity, regulating cellular replicative life span. Telomerase is not expressed in most normal human somatic cells but is active in stabilizing telomeres of certain self-renewing cell populations and most malignant cells, making the enzyme an appealing target for anticancer therapy. We describe here a novel cross-species approach to telomerase inhibition. Ectopic expression of the human telomerase catalytic reverse transcriptase component in murine cells inhibited endogenous murine telomerase activity. Using this approach, telomerase inhibition in immortal murine fibroblasts resulted in critical telomere shortening, leading to slowed proliferation, abnormal morphology, altered cell cycle, and telomere dysfunction with cytogenetic instability, followed by apoptotic cell death. Subpopulations of two telomerase-inhibited clones escaped widespread apoptosis, showing proliferative recovery in culture despite persistently inhibited telomerase activity with progressive telomere shortening and dysfunction. This study, by targeting immortal murine cells for telomerase inhibition, demonstrates the importance of telomerase to murine cell immortalization and telomere maintenance. Moreover, the murine model used here should prove useful in further evaluating telomerase inhibition as an anticancer therapy. PMID- 11929833 TI - Rad6 overexpression induces multinucleation, centrosome amplification, abnormal mitosis, aneuploidy, and transformation. AB - We have isolated by differential RNA display a cDNA that is up-regulated in metastatic mammary tumor lines. This cDNA corresponds to HR6B, the yeast homologue of Rad6, a ubiquitin-conjugating enzyme, and a key player in postreplication repair and induced mutagenesis in the yeast. We show that Rad6 protein expressed in metastatic tumor lines is wild type and functional, because it is able to catalyze the transfer of ubiquitin to histone H2b and is predominantly localized in the nucleus as compared with cytoplasmic localization in normal or nonmetastatic mammary cells. This pattern of Rad6 protein expression/localization is not restricted to breast cancer cell lines, because human breast carcinomas display similar patterns of Rad6 up-regulation and nuclear localization suggesting that deregulation in expression of Rad6 may be an important step in transformation to malignant phenotype. Constitutive overexpression of exogenous human HR6B cDNA into normal-behaving MCF10A human breast epithelial cells induced cell-cell fusion that resulted in generation of multinucleated cells, centrosome amplification, multipolar mitotic spindles, aneuploidy, and ability for anchorage-independent growth. Double immunofluorescence labeling experiments demonstrated the colocalization of Rad6 protein with gamma-tubulin on centrosomes. This physical association of Rad6 with centrosomes is maintained throughout the interphase and mitotic phases of the cell cycle. The Rad6 protein exhibits notable alterations in distribution during interphase and mitotic stages of the cell cycle that are compatible with its function as a transcription factor. These findings suggest that Rad6 is an important ubiquitin-conjugating enzyme that may play a significant role in the maintenance of genomic integrity of mammalian cells and that an imbalance in the levels and activity of Rad6 could lead to chromosomal instability and transformation in vitro. PMID- 11929834 TI - Collective cell movement in primary melanoma explants: plasticity of cell-cell interaction, beta1-integrin function, and migration strategies. AB - Collective cell movement represents an efficient dissemination strategy in neoplastic epithelial and mesenchymal cancer. In primary melanoma explants cultured in three-dimensional collagen lattices, invasive migration of multicellular clusters was dependent on the function of beta1 integrins, as shown by preferential beta1-integrin expression and clustering in a subset of promigratory cells at the leading edge ("guiding cells") and the abrogation of multicellular migration by adhesion-perturbing anti-beta1-integrin antibody. Interference with beta1-integrin function induced complex changes in cluster polarity and cohesion, including development of two or several opposing leading edges, cluster disruption, and the detachment of individual cells followed by beta1-integrin-independent "amoeboid" crawling and dissemination. The conversion from beta1-integrin-dependent collective movement to beta1-integrin-independent single-cell motility suggests efficient cellular and molecular plasticity in tumor cell migration strategies. PMID- 11929835 TI - Suppression of Rac activity induces apoptosis of human glioma cells but not normal human astrocytes. AB - Tumors of glial origin such as glioblastoma multiforme (GBM) comprise the majority of human brain tumors. Patients with GBM have a very poor survival rate, with an average life expectancy of <1 year. We asked whether we could identify a survival pathway in high-grade glioma and oligodendroglioma cells that when suppressed, would induce apoptosis of these tumor cells but not of normal human adult astrocytes. To identify these pathways, we selectively suppressed the activity of a number of proteins (Ras, Rac1, Akt1, RhoA, c-jun, and MEK1/2) hypothesized to play roles in cell survival. We found that suppression of Rac1, a small GTP-binding protein, inhibited survival and produced apoptosis in three human glioma cell lines (U87, U343, and U373). Serum induced the activity of Rac1 and the activity or phosphorylation state of p21-activated kinase 1 and c-Jun NH(2)-terminal kinase (JNK), two intracellular targets of Rac1. Suppression of Rac1 also induced apoptosis in 19 of 21 short-term cultures of human primary cells from grades II and III oligodendroglioma and grade IV glioblastoma that varied in p53, epidermal growth factor receptor, epidermal growth factor receptor vIII, MDM2, and p16/p19 mutational or amplification status. In contrast, inhibition of Rac1 activity did not induce apoptosis of normal primary human adult astrocytes. In both established glioma cell lines and primary glioma cells, apoptosis induced by the inhibition of Rac was partially rescued by activated mitogen-activated protein kinase kinase 1, an activator of JNK, suggesting that JNK functions downstream of Rac1 in glioma cells. These results indicate that Rac1 regulates a major survival pathway in most glioma cells, and that suppression of Rac1 activity stimulates the death of virtually all glioma cells, regardless of their mutational status. Agents that suppress Rac1 activity may therefore be useful therapeutic treatments for malignant gliomas. PMID- 11929836 TI - Human non-small cell lung tumors and cells derived from normal lung express both estrogen receptor alpha and beta and show biological responses to estrogen. AB - Lung cancer is becoming increasingly common in women and in the United States accounts for more female cancer deaths annually than breast cancer. Many epidemiological studies have provided evidence that women are more susceptible than men to the adverse effects of tobacco smoke. These observations suggest the possible role of estrogens in lung carcinogenesis. We report here the expression of mRNA for estrogen receptor alpha (ERalpha) and beta (ERbeta) in cultured human non-small cell lung cancer cells, cultured lung fibroblasts, and primary cultures of normal bronchial epithelium. Western analysis of ERalpha suggested that the main protein expressed in lung tumor cells is a variant, probably attributable to alternative splicing. Protein for ERbeta was found to be a mixture of full-length as well as alternatively spliced variants. beta-Estradiol produced a proliferative response in vitro in both normal lung fibroblasts and cultured non small cell lung tumor cells. This effect was also observed in vivo. In this regard, beta-estradiol stimulated growth of the non-small cell lung tumor line, H23, grown as tumor xenografts in SCID mice. This effect was blocked by fluvestrant (ICI 182,780). In paraffin sections of non-small cell lung tumors, ERbeta immunoreactivity was localized to the nucleus, whereas ERalpha immunoreactivity was mainly localized to the cytoplasm, suggesting that both nuclear and cytoplasmic signaling may be involved in estrogenic responses in the lung. To show that the ERs found in the lung are functional, we demonstrated that beta-estradiol stimulated transcription of an estrogen response element luciferase construct transfected in non-small cell lung tumor cell lines. Antiestrogens blocked this effect. Treatment of lung fibroblasts with beta estradiol also increased secretion of hepatocyte growth factor by 2-fold. These results suggest that estrogen signaling plays a biological role in both the epithelium and the mesenchyme in the lung and that estrogens could potentially promote lung cancer, either through direct actions on preneoplastic or neoplastic cells or through indirect actions on lung fibroblasts. Additionally, it is possible that antiestrogens may have therapeutic value to treat or prevent lung cancer. PMID- 11929837 TI - Vascular accumulation of a novel photosensitizer, MV6401, causes selective thrombosis in tumor vessels after photodynamic therapy. AB - The antivascular effects of photodynamic therapy (PDT) and their mechanisms are not clearly understood. Here, we examined the effects of PDT with a novel photosensitizer MV6401 on the microvasculature in a mammary tumor (MCaIV) grown in a murine dorsal skinfold chamber and in normal tissue controls. The mice were irradiated with light 15 min after i.v. administration of MV6401 when the drug was localized only in the vascular compartment, as shown by fluorescence microscopy and immunohistochemistry. PDT with MV6401 caused a dose-dependent biphasic blood flow stasis and vascular hyperpermeability, as determined by intravital microscopy. This biphasic response was classified into two components: (a) an acute response observed immediately after PDT; and (b) a long-term response observed at times greater than 3 h after PDT. The acute temporal vascular effects were characteristic of vasoconstriction but not of thrombus formation. However, the long-term vascular shutdown was mediated by thrombus formation, as evidenced by histological evaluation and inhibition with heparin. Minimal effects were observed in normal vessels after antivascular doses used against the tumor, but there was no long-term vascular damage. In concert with the stasis, a dose-dependent tumor growth delay was observed. This study provides mechanistic insights into antitumor vascular effects of PDT and suggests novel strategies for tumor treatment with PDT. PMID- 11929839 TI - Persistence of solitary mammary carcinoma cells in a secondary site: a possible contributor to dormancy. AB - Tumors can recur years after treatment, and breast cancer is especially noted for long periods of dormancy. The status of the cancer during this period is poorly understood. As a model to study mechanisms of dormancy, we used murine D2.0R mammary carcinoma cells, which are poorly metastatic but form occasional metastases in liver and other organs after long latency. Highly metastatic D2A1 cells provided a positive, metastatic control. Our goals were to learn how the cell lines differ in survival kinetics in a secondary site and to seek evidence for the source of D2.0R dormancy. In spontaneous metastasis assays from mammary fat pad injections, we found evidence for dormancy because of a persistence of large numbers of solitary cells in the liver. To quantify the fate of cells after arrival in liver, experimental metastasis assays were used. To permit identification of cells that had not divided, cells were labeled before injection with fluorescent nanospheres, which were diluted to undetectable levels by cell division. Cancer cells were injected i.v. to target them to the liver and coinjected with reference microspheres to monitor cell survival. Dormancy was defined as retention of nanosphere fluorescence in vivo, as well as negative staining for the proliferation marker Ki67. A large proportion of D2.0R cells persisted as solitary dormant cells. No metastases formed, but viable cells could be recovered from the liver 11 weeks after injection. Large numbers of solitary, dormant, Ki67-negative D2A1 cells were also detected against a background of progressively growing metastases. Thus, this study identified a possible contributor to tumor dormancy: solitary, dormant cells that persist in tissue. If such cells are present in patients, they could contribute to tumor recurrence and would not be susceptible to current therapeutic strategies targeting proliferating cells. PMID- 11929838 TI - Tumor-specific down-regulation of the tumor necrosis factor-related apoptosis inducing ligand decoy receptors DcR1 and DcR2 is associated with dense promoter hypermethylation. AB - TRAIL (tumor necrosis factor-related apoptosis-inducing ligand) induces apoptosis in a large variety of cancer cells but not in most normal human cells. This feature makes TRAIL, a potential antitumor agent. TRAIL can bind to four different receptors, two pro-apoptotic death receptors (DRs), DR4 and DR5, and two antiapoptotic decoy receptors (DcRs), DcR1 and DcR2. Normal cells express all four of the receptors. The increased TRAIL sensitivity of tumor cells has been postulated to result from the lack of DcR expression. We studied the tumor specific down-regulation of the TRAIL receptors DcR1 and DcR2, as well as DR4 and DR5, in a group of pediatric tumor cell lines [nine neuroblastoma and three peripheral primitive neuro-ectodermal tumors (PNETs)] and three cell lines from adult tumors. Lack of expression of DcR1 and DcR2 was widespread (13 of the 15 cell lines and 10 of 15, respectively), both in the adult tumor cell lines and in the pediatric tumor lines. DR4 and DR5 were expressed in 8 of 15 and 12 of 15 cell lines, respectively. To understand the tumor-specific down-regulation of the TRAIL receptors, the promoter regions were studied for possible methylation changes of their CpG islands. All normal tissues were completely unmethylated, whereas in the tumor cell lines, we found frequent hypermethylation of the promoter. For DcR1 and DcR2, we found dense hypermethylation in 9 (69%) of 13 and 9 (90%) of 10 of nonexpressing cell lines, respectively. DR4 and DR5 were methylated in 5 (71%) of 7 and 2 (67%) of 3 nonexpressing cell lines, respectively. Treatment with the demethylating agent 5-aza-2'deoxycytidine resulted in partial demethylation and restored mRNA expression. In addition, we performed mutation analysis of the death domains of DR4 and DR5 by sequencing exon 9. Mutations were not present in any of the neuroblastoma or PNET cell lines. A panel of 28 fresh neuroblastoma tumor samples also lacked expression of DcR1 and DcR2 in 85 and 74% of cases, respectively. Hypermethylation was observed in 6 (21%) of 28 for DcR1 and 7 (25%) of 28 for DcR2. DR4 and DR5 were both expressed in 22 of 28 tumors, and no promoter methylation was observed. These data suggest that hypermethylation of the promoters of DcR1 and DcR2 is important in the down-regulation of expression in neuroblastoma and other tumor types. PMID- 11929840 TI - Growth phase-dependent expression of ICAD-L/DFF45 modulates the pattern of apoptosis in human colonic cancer cells. AB - The inhibitor of caspase-3-activated DNase (ICAD) is a caspase-3 substrate that controls nuclear apoptosis. ICAD has two isoforms: a functional isoform of M(r) 45,000, ICAD-L/DNA fragmentation factor (DFF) 45; and a M(r) 35,000 isoform, ICAD S/DFF35. ICAD-deficient murine cells display resistance to apoptotic stimuli and absence of typical nuclear changes of apoptosis. Our aim was to: (a) characterize the ICAD expression in several human colonic cancer cell lines compared with human normal colonocytes; and (b) correlate the phenotypic features of apoptosis to the level of ICAD expression. ICAD expression was assessed by immunoblot analysis. Early markers of apoptosis of cultured cells included lactate dehydrogenase retention in dying cells, cytokeratin 18 cleavage, and caspase-3 activation. Nuclear markers of apoptosis were assessed by Hoechst staining of nuclei, electron microscopy, and DNA electrophoresis. Inhibition of caspases was performed using a broad-spectrum caspase inhibitor, z-Val-Ala-Asp-fluoromethyl ketone. ICAD expression was restricted to the functional ICAD-L/DFF45 isoform in colonic cancer cells as well as in human normal colonocytes. In a clonal derivative of HT29 cells (HT29-Cl.16E cells), ICAD expression was found to be down-regulated during the exponential phase of growth, and the cell death triggered by IFN-gamma, anti-Fas antibody plus Adriamycin was characterized by the expression of early markers of apoptosis, whereas the key nuclear features of apoptosis were absent. In contrast, exposure of confluent cells to this treatment led to a typical apoptotic nuclear fragmentation. Both forms of apoptosis, in exponentially growing and confluent cells, were sensitive to the broad spectrum inhibitor of caspases, z-Val-Ala-Asp-fluoromethyl ketone. Our findings support the concept that the expression of ICAD is essential to the execution of full blown apoptosis in colonic cancer cells. Altogether, our results point to ICAD as a potential target for restoring a normal apoptotic signal transduction pathway in colonic cancer cells. PMID- 11929841 TI - Antisense RNA down-regulation of bcl-xL Expression in prostate cancer cells leads to diminished rates of cellular proliferation and resistance to cytotoxic chemotherapeutic agents. AB - bcl-xL is a M(r) 26,000 bcl-2 homologue that is highly expressed in prostate cancer cells. In previous studies, the down-regulation of its expression by antisense oligonucleotides led to resistance. In this work, the 445-bp 5' terminus of the bcl-xL cDNA was cloned in the antisense orientation and stably transfected into DU145 and LNCaP prostate cancer cells. In the DU145 (and to a lesser extent the LNCaP) transfectants, phenotypic changes (versus mock transfected cells) included an increase in doubling time (from 36 to 175 h) in the clone in which bcl-xL protein expression was 25% of control. The transfectants did not demonstrate characteristic apoptotic changes, as demonstrated by 4',6-diamidino-2-phenylindole staining, lack of either DNA laddering, caspase-3 activation, or poly(ADP)ribose and lamin cleavage, and the absence of a significant sub-G(0) population. Cell cycle analysis demonstrated an increase in a tetraploid population (from 28% to 66%), as well as the appearance of a hypertetraploid population. Levels of cIAP-1 protein were almost undetectable in the mock cells but increased at least 25-fold in the DU145 transfectants. The down-regulation of bcl-xL in both DU145 (and to a much lesser extent in LNCaP) cells led to their resistance to cytotoxic agents, including docetaxel, mitoxantrone, etoposide, vinblastine, and carboplatin. Reversion of bcl-xL expression in stable DU145 transfectants to nearly the levels found in the mock-transfected cells was accomplished by retroviral infection of the cells with a bcl-xL sense cDNA under control of a prolific promoter. This led to a dramatic increase in the growth rate and in BrdUrd incorporation, as well as a sharp decrease in the expression of cIAP-1 protein. Overall, these findings highlight the adaptability of prostate cancer cells to loss of bcl-xL and suggest that in addition to its prosurvival role, bcl-xL protein may also be involved in the regulation of the rate of cellular proliferation. PMID- 11929842 TI - Loss of Bax alters tumor spectrum and tumor numbers in ARF-deficient mice. AB - p19(ARF) is a key regulator of the p53-mediated apoptotic and tumor suppressor pathway. The proapoptotic Bax gene is a transcription target of p53, yet genetic studies in some animal models have suggested that Bax and p53 loss may cooperate in tumorigenesis. ARF-deficient mice are tumor prone, and to determine whether Bax loss could cooperate in the development of these tumors, we generated mice null for both ARF and Bax. The tumor latency of Bax+/+ARF-/-, Bax+/-ARF-/- and Bax-/-ARF-/- mice was similar with a mean survival of 48.9, 48.1, and 47.6 weeks, respectively. In Bax+/+ARF-/- mice, the predominant tumor type was B- and T-cell lymphoma followed by sarcomas and a lack of carcinomas. However, the frequency of lymphoma development dramatically decreased, whereas that of sarcomas and carcinomas increased, in a gene dosage-dependent manner in Bax+/-ARF-/- and Bax-/ ARF-/- mice. Furthermore, uncommon tumors of ARF-/- mice (osteosarcoma and hemangiosarcoma) were observed in Bax/ARF-double null mice, and tumor types not described previously in ARF-null mice (mixed germ cell tumor, Triton tumor, and histiocytic sarcoma) also developed in Bax-/-ARF-/- animals. Importantly, multiple primary malignant tumors of different lineage arose in 25% of the Bax-/ ARF-/- mice, whereas only one tumor type per animal was observed in Bax+/+ARF null littermates. Finally, the wild-type Bax allele was retained in tumors arising in Bax+/-ARF-/- mice. Thus, Bax appears to function as a tumor modifier rather than as a classic tumor suppressor, and the combined loss of Bax and the ARF allows for the emergence of multiple malignant tumor types, an alteration of the tumor spectrum, and tumors not observed previously in ARF-null mice. PMID- 11929843 TI - Correspondence re: A. Kumar et al., cross-resistance to the synthetic retinoid CD437 in a paclitaxel-resistant human ovarian carcinoma cell line is independent of the overexpression of retinoic acid receptor-gamma. Cancer Res., 61: 7552 7555, 2001. PMID- 11929844 TI - Special conference of the American Association for Cancer Research on molecular imaging in cancer: linking biology, function, and clinical applications in vivo. AB - The AACR Special Conference on Molecular Imaging in Cancer: Linking Biology, Function, and Clinical Applications In Vivo, was held January 23-27, 2002, at the Contemporary Hotel, Walt Disney World, Orlando, FL. Co-Chairs David Piwnica Worms, Patricia Price and Thomas Meade brought together researchers with diverse expertise in molecular biology, gene therapy, chemistry, engineering, pharmacology, and imaging to accelerate progress in developing and applying technologies for imaging specific cellular and molecular signals in living animals and humans. The format of the conference was the presentation of research that focused on basic and translational biology of cancer and current state-of the-art techniques for molecular imaging in animal models and humans. This report summarizes the special conference on molecular imaging, highlighting the interfaces of molecular biology with animal models, instrumentation, chemistry, and pharmacology that are essential to convert the dreams and promise of molecular imaging into improved understanding, diagnosis, and management of cancer. PMID- 11929845 TI - Oestrogenic repression of human coagulation factor VII expression mediated through an oestrogen response element sequence motif in the promoter region. AB - Reporter gene analysis of two regions of the human factor VII (FVII) gene promoter (residues -658 to -1 and -348 to -1, where +1 is the start site of translation) in the mammalian liver-derived cell line HepG2 showed reduced transcriptional activity in the presence of oestrogenic factors. This effect was independent of promoter polymorphic haplotype. Similar analysis using a smaller region of the promoter spanning residues -187 to -1 failed to show any evidence of oestrogenic suppression. Electrophoretic mobility shift assays and supershift assays using recombinant oestrogen receptor alpha and anti-oestrogen receptor antibody localized the sequence motif to which oestrogen receptor was binding to residues -225 to -212 of the FVII promoter. The lack of oestrogenic suppression in a reporter gene construct spanning residues -658 to -1 modified to abolish oestrogen receptor binding at this site, confirmed the functional significance of this motif. Although superficially similar to the classical oestrogen response element (ORE), comprising two half sites separated by three spacer nucleotides, the FVII ORE represents an alternative type of ORE in which the two half sites are separated by just two spacer nucleotides. EMSAs indicated that increasing spacer nucleotide number from two to three in the FVII ORE, or decreasing it from three to two in a consensus ORE sequence motif, had a small effect on the binding affinity for oestrogen receptor. These data correlate with and provide a plausible mechanism for the inverse relationship between FVII and oestradiol levels observed during the menstrual cycle. PMID- 11929846 TI - Adeno-associated virus vector gene transfer and sarcolemmal expression of a 144 kDa micro-dystrophin effectively restores the dystrophin-associated protein complex and inhibits myofibre degeneration in nude/mdx mice. AB - Duchenne muscular dystrophy is a severe life-threatening X-linked recessive disorder, caused by mutations in the dystrophin gene, for which currently there is no effective treatment. Because of the large size of the dystrophin cDNA (14 kb) this precluded it from being used in early adenovirus- or retrovirus-based gene therapy vectors. However, some therapeutic success has been achieved in mdx mice using adenovirus- and retrovirus-mediated transfer of a 6.3 kb recombinant mini-dystrophin cDNA. Despite this, problems with immunogenicity and inefficient transduction of mature myofibres make these vectors less than ideal for gene transfer to skeletal muscle. Adeno-associated viral (AAV) vectors overcome many of the problems associated with other vector systems. However, AAV vectors can only accommodate <5 kb of foreign DNA. For this reason we have produced a micro dystrophin cDNA gene construct that is <3.8 kb. This construct, driven by a CMV promoter, was introduced into the skeletal muscle of 12-day-old nude/mdx mice using an AAV vector, resulting in specific sarcolemmal expression of micro dystrophin in >50% of myofibres up to 20 weeks of age, and effective restoration of the dystrophin-associated protein (DAP) complex components. Additionally, evaluation of central nucleation indicated a significant inhibition of degenerative dystrophic muscle pathology. We have therefore shown that the current micro-dystrophin gene delivered in vivo using an AAV vector is not only capable of restoring sarcolemmal DAP complexes, but can also ameliorate dystrophic pathology at the cellular level. PMID- 11929847 TI - A molecular basis for differential developmental anomalies in Axenfeld-Rieger syndrome. AB - Pitx2, a bicoid-like homeodomain transcription factor and Dlx2 are two transcriptional markers observed during early tooth development. PITX2 binds to bicoid and bicoid-like elements in the Dlx2 promoter and activates this promoter 30-fold in Chinese hamster ovary cells. Mutations in PITX2 associated with Axenfeld-Rieger syndrome (ARS) provided the first link of this homeodomain transcription factor to tooth development. We are investigating the molecular basis of developmental anomalies associated with human PITX2 mutations. A phenotypically less severe ARS mutant (without tooth anomalies), PITX2 R84W, has a similar DNA binding specificity compared to wild-type PITX2 and transactivates the Dlx2 promoter. This mutation is associated with iris hypoplasia (IH); in contrast a Rieger syndrome mutation, PITX2 T68P, which presents clinically with the full spectrum of developmental anomalies (including tooth anomalies), is unable to transactivate the Dlx2 promoter. Since Dlx2 expression is required for tooth and craniofacial development the lack of tooth anomalies in the patient with IH may be due to the residual activity of this mutant in activating the Dlx2 promoter. We demonstrate that PITX2 phosphorylation increases PITX2 and PITX2 R84W DNA binding. The PITX2 T68P ARS mutation occurs at a protein kinase C phosphorylation site in the homeodomain. Surprisingly, phosphorylation of PITX2 T68P is increased compared to wild-type PITX2 but has little effect on its DNA binding activity. Altogether these data suggest a molecular mechanism for tooth development involving Dlx2 gene expression in ARS patients. PMID- 11929848 TI - Novel ENU-induced eye mutations in the mouse: models for human eye disease. AB - We have carried out a genome-wide screen for novel N-ethyl-N-nitrosourea-induced mutations that give rise to eye and vision abnormalities in the mouse and have identified 25 inherited phenotypes that affect all parts of the eye. A combination of genetic mapping, complementation and molecular analysis revealed that 14 of these are mutations in genes previously identified to play a role in eye pathophysiology, namely Pax6, Mitf, Egfr and Pde6b. Many of the others are located in genomic regions lacking candidate genes and these define new loci. Four of the mutants display a similar phenotype of dilated pupils but do not appear to be allelic, and at least two of these are embryonic lethal when homozygous. This collection of eye mutations will be valuable for understanding gene function, for dissecting protein function and as models of human eye disease. PMID- 11929849 TI - A novel interaction between lamin A and SREBP1: implications for partial lipodystrophy and other laminopathies. AB - The gene encoding nuclear lamins A and C is mutated in at least three inherited disorders. Two of these, Emery-Dreifuss muscular dystrophy (EDMD-AD) and a form of dilated cardiomyopathy (CMD1A), involve muscle defects, and the other, familial partial lipodystrophy (FPLD), involves loss of subcutaneous adipose tissue. Mutations causing FPLD, in contrast to those causing muscle disorders, are tightly clustered within the C-terminal domain of lamin A/C. We investigated the expression and subcellular localization of FPLD lamin A mutants and found no abnormalities. We therefore set out to identify proteins interacting with the C terminal domain of lamin A by screening a mouse 3T3-L1 adipocyte library in a yeast two-hybrid interaction screen. Using this approach, the adipocyte differentiation factor, sterol response element binding protein 1 (SREBP1) was identified as a novel lamin A interactor. In vitro glutathione S-transferase pull down and in vivo co-immunoprecipitation studies confirmed an interaction between lamin A and both SREBP1a and 1c. A binding site for lamin A was identified in the N-terminal transcription factor domain of SREBP1, between residues 227 and 487. The binding of lamin A to SREBP1 was noticeably reduced by FPLD mutations. Interestingly, one EDMD-AD mutation also interfered with the interaction between lamin A and SREBP1. Whilst the physiological relevance of this interaction has yet to be elucidated, these data raise the intriguing possibility that fat loss seen in laminopathies may be caused, at least in part, by reduced binding of the adipocyte differentiation factor SREBP1 to lamin A. PMID- 11929850 TI - Heteroligomerization of an Aquaporin-2 mutant with wild-type Aquaporin-2 and their misrouting to late endosomes/lysosomes explains dominant nephrogenic diabetes insipidus. AB - Autosomal nephrogenic diabetes insipidus (NDI), a disease in which the kidney is unable to concentrate urine in response to vasopressin, is caused by mutations in the Aquaporin-2 (AQP2) gene. Analysis of a new family with dominant NDI revealed a single nucleotide deletion (727deltaG) in one AQP2 allele, which encoded an AQP2 mutant with an altered and extended C-terminal tail. When expressed in oocytes, the tetrameric AQP2-727deltaG was retained within the cell. When co expressed, AQP2-727deltaG, but not a mutant in recessive NDI (AQP2-R187C), formed hetero-oligomers with wild-type (wt) AQP2 and reduced the water permeability of these oocytes, because of a reduced plasma membrane expression of wt-AQP2. Expressed in renal epithelial cells, AQP2-727deltaG predominantly localized to the basolateral membrane and late endosomes/lysosomes, whereas wt-AQP2 was expressed in the apical membrane. Upon co-expressing in these cells, wt-AQP2 and AQP2-727deltaG mainly co-localized to late endosomes/lysosomes. In conclusion, hetero-oligomerization of AQP2-727deltaG with wt-AQP2 and consequent mistargeting of this complex to late endosomes/lysosomes results in absence of AQP2 in the apical membrane, which can explain dominant NDI in this family. Together with other mutants in dominant NDI, our data reveal that a misrouting, instead of a lack of function, is a general mechanism for the 'loss of function' phenotype in dominant NDI and visualizes for the first time a mislocalization of a wild-type protein to late endosomes/lysosomes in polarized cells after oligomerization with a mutant protein. PMID- 11929851 TI - Age and insertion site dependence of repeat number instability of a human DM1 transgene in individual mouse sperm. AB - Precise measurement of germline repeat number mutations is important for understanding the molecular etiology of expanded trinucleotide repeat diseases. We used single genome-equivalent PCR of sperm DNA to measure the mutation frequencies in two lines of Dmt transgenic mice containing an expanded CTG.CAG tract on an identical genetic background. Single genome-equivalent PCR indicated that apparent mutational spectra derived in other investigations from PCR of bulk sperm DNA were largely the consequence of PCR stutter and not mutations. Here we show that sperm from 8-week-old Dmt-D mice had a significantly higher mutation frequency (change of >1 repeat) (14.2%) than those of Dmt-E mice of the same age (5.5%), in agreement with pedigree analysis. Furthermore, the mutation frequency in sperm of Dmt-D mice increased significantly with age (28.0% at 17 weeks). The age dependence of the degree of expansion implies that mutations accumulate with time in spermatogenic stem cells. Similar rates of expansion per spermatogenic cycle in man would yield the large expansions observed in human diseases such as myotonic dystrophy type 1. Pedigree data showed a significant age-dependent bias toward repeat contraction in female transmissions and a trend towards expansion with age in male transmissions. Thus, direct single genome-equivalent PCR of the sperm DNA of an individual male appears to predict the distribution of mutant allele sizes that might be inherited by its offspring. In further contrast to a recent report, the sex of the offspring had no detectable effect on the direction of the mutational length change. PMID- 11929852 TI - Gain-of-function mutation in ADULT syndrome reveals the presence of a second transactivation domain in p63. AB - The transcriptional co-activator p63 is of crucial importance for correct development of the limbs, ectodermal appendages (skin, nails, teeth, hair, glands), lip and palate. Mutations in the p63 gene are found in a number of human syndromes, including ectrodactyly-ectodermal dysplasia-cleft lip/palate (EEC) syndrome, limb-mammary syndrome (LMS), Hay-Wells syndrome and in non-syndromic split-hand/split-foot malformation (SHFM). Each syndrome has a specific pattern of mutations with different functional effects in in vitro functional assays. We report a mutation R298Q in acro-dermato-ungual-lacrimal-tooth (ADULT) syndrome, another EEC-like condition. The mutation is located in the DNA binding domain of p63, which harbors almost all EEC associated mutations. However, unlike mutations in EEC syndrome, the R298Q ADULT syndrome mutation does not impair DNA binding. Rather, the mutation confers novel transcription activation capacity on the DeltaN-p63gamma isoform, which normally does not possess such activity. These results confirm that ADULT syndrome is a clinically as well as molecularly distinct member of the expanding p63 mutation family of human malformation syndromes. Our results further show that p63 contains a second transactivation domain which is normally repressed and can become activated by mutations in the DNA binding domain of p63. PMID- 11929853 TI - Three proteins, MBNL, MBLL and MBXL, co-localize in vivo with nuclear foci of expanded-repeat transcripts in DM1 and DM2 cells. AB - Myotonic dystrophy is a complex neuromuscular disorder associated with DNA expansion mutations in two different genes. In DM1 a CTG repeat in the 3' untranslated region of DMPK is expanded, whereas in DM2 an intronic CCTG expansion occurs in the gene ZNF9. Transcripts containing expanded repeats form foci in the nuclei of DM1 and DM2 cells. Recent work using antibodies has shown that proteins related to Drosophila muscleblind co-localize with repeat foci in DM1 and DM2 cells. We show that rather than there being a single human muscleblind gene producing multiple proteins through alternative splicing, there are in fact three different muscleblind genes, MBNL, MBLL and MBXL, which map to chromosomes 3, 13 and X, respectively, and which show extensive alternative splicing. Two of the genes, MBNL and MBLL, are expressed in many adult tissues whereas MBXL is expressed predominantly in the placenta. Green fluorescent protein-tagged versions of MBNL, MBLL and MBXL co-localize with nuclear foci in DM1 and DM2 cells, suggesting that all three proteins may play a role in DM pathophysiology. PMID- 11929855 TI - The Leber congenital amaurosis gene product AIPL1 is localized exclusively in rod photoreceptors of the adult human retina. AB - Leber congenital amaurosis (LCA) is the most severe inherited retinal dystrophy resulting in markedly impaired vision or blindness at birth. LCA is characterized by an extinguished electroretinogram in infancy, which is thought to be indicative of an early and severe impairment of both the rod and cone photoreceptors in the human retina. Recently, the aryl hydrocarbon receptor interacting protein-like 1 (AIPL1) gene was identified as the fourth causative gene of LCA. AIPL1 encodes a 384 amino acid protein of unknown function. We have generated a polyclonal antibody against a peptide from a unique region within the primate AIPL1 protein, which detects a protein of approximately 43 kDa in human retinal extracts. A screen of human tissues and immortalized cell lines with this antibody reveals AIPL1 to be specific to human retina and cell lines of retinal origin (Y79 retinoblastoma cells). Within the retina, AIPL1 was detected only in the rod photoreceptor cells of the peripheral and central human retina. The AIPL1 staining pattern extended within the rod photoreceptor cells from the inner segments, through the rod nuclei to the rod photoreceptor synaptic spherules in the outer plexiform layer. AIPL1 was not detected in the cone photoreceptors of peripheral or central human retina. This study is the first to suggest that AIPL1 performs a function essential to the maintenance of rod photoreceptor function. PMID- 11929854 TI - Frataxin promotes antioxidant defense in a thiol-dependent manner resulting in diminished malignant transformation in vitro. AB - Friedreich ataxia is an inherited disorder caused by decreased expression of frataxin protein. Increasing evidence suggests that this protein might detoxify reactive oxygen species (ROS) by an unknown mechanism. Here we demonstrate that transgenic overexpression of human frataxin increases cellular antioxidant defense via activation of glutathione peroxidase and elevation of reduced thiols, thereby reducing the incidence of malignant transformation induced by ROS, as observed by soft agar assays and tumour formation in nude mice. These findings expand the understanding of antioxidant properties of frataxin, and tentatively suggest a role in the early induction of cancer. PMID- 11929856 TI - Lipoid proteinosis maps to 1q21 and is caused by mutations in the extracellular matrix protein 1 gene (ECM1). AB - Lipoid proteinosis (LP), also known as hyalinosis cutis et mucosae or Urbach Wiethe disease (OMIM 247100) is a rare, autosomal recessive disorder typified by generalized thickening of skin, mucosae and certain viscera. Classical features include beaded eyelid papules and laryngeal infiltration leading to hoarseness. Histologically, there is widespread deposition of hyaline (glycoprotein) material and disruption/reduplication of basement membrane. The aetiology of LP is currently unknown. Using DNA from three affected siblings in a consanguineous Saudi Arabian family we performed genome-wide linkage and mapped the disorder to 1q21 (marker D1S498) with a two-point LOD score of 3.45 at theta = 0. A further 28 affected individuals from five other unrelated consanguineous family groups from different geographical regions also showed complete linkage and resulted in a maximum two-point LOD score of 21.85 at theta = 0. Using available markers in the interval between D1S442 and D1S305, the observed recombinants placed the gene in a 2.3 cM critical interval between D1S2344 and D1S2343 (Marshfield genetic map) corresponding to an approximately 6.5 Mb region on the UCSC physical map. Using a candidate gene approach (comparison of control versus LP gene expression in cultured fibroblasts) and subsequent direct sequencing of genomic DNA, we identified six different homozygous loss-of-function mutations in the extracellular matrix protein 1 gene (ECM1). Although the precise function of ECM1 is not known, our findings provide the first clinical indication of its relevance to skin adhesion, epidermal differentiation, wound healing, scarring, angiogenesis/angiopathy and basement membrane physiology, as well as defining the molecular basis of this inherited disorder. PMID- 11929857 TI - Structural analysis of the chicken BRCA2 gene facilitates identification of functional domains and disease causing mutations. AB - Carriers of mutations in the BRCA2 gene have a high risk of developing breast and other cancers. The BRCA2 gene, which is located on human chromosome 13, encodes a very large protein of only poorly understood function. To define regions of sequence conservation and highlight potentially functionally important domains, we have cloned and characterized the chicken BRCA2 gene, the first non-mammalian BRCA2 gene to be described. The gene is organized similarly to the human BRCA2 gene, but is more compact and is localized to the subtelomeric region of chicken chromosome 1q, within a region that contains other genes from human chromosome 13. The chicken BRCA2 gene encodes a protein of 3399 amino acids, which is poorly conserved with mammalian BRCA2 proteins, having only 37% amino acid identity overall with human BRCA2. However, certain domains are much more highly conserved, indicating functional significance. We describe genes with some of these conserved domains in organisms as diverse as intracellular parasites, mosquitoes and plants. The evolutionarily divergent chicken BRCA2 sequence may also be useful in assigning the large number of sequence variants that have been described in the human BRCA2 gene which are of unknown significance in disease causation. PMID- 11929858 TI - Hyperekplexia associated with compound heterozygote mutations in the beta-subunit of the human inhibitory glycine receptor (GLRB). AB - Hyperekplexia (MIM: 149400) is a neurological disorder characterized by an excessive startle response which can be caused by mutations in the alpha1-subunit (GLRA1) of the heteropentameric human inhibitory glycine receptor (hGlyR). These receptors facilitate fast-response, inhibitory glycinergic neurotransmission in the brainstem and spinal cord leading to a rapid modification and reduction of the excitatory startle response. Mutations in the beta-subunit of GlyR (glrb) occur in a murine model of hyperekplexia (spastic), but have not been detected in human hyperekplexia. Following mutation analysis of the human beta-subunit of hGlyR (GLRB) in a cohort of 22 hyperekplexia patients, we provide evidence to confirm that GLRB mutations can cause human hyperekplexia. A missense (G920A resulting in G229D) and a splice site mutation (IVS5+5G-->A) occurred together in a compound heterozygote with a transient hyperekplexia phenotype. Exon trap analysis revealed that IVS5+5G-->A results in the exclusion of exon 5 from GLRB transcripts. Electrophysiological studies showed reduced sensitivity to agonist mediated activation of the alpha1beta (G229D) GlyR suggesting that GlyR beta subunits are not restricted to conferring modulatory influences and maintaining structural integrity, but may also play a functional role in hGlyR ligand binding. PMID- 11929859 TI - Complexin regulates the closure of the fusion pore during regulated vesicle exocytosis. AB - Membrane fusion during exocytosis and throughout the cell is believed to involve members of the SNARE (soluble N-ethylmaleimide-sensitive fusion protein attachment protein receptors) family of proteins. The assembly of these proteins into a four-helix bundle may be part of the driving force for bilayer fusion. Regulated exocytosis in neurons and related cell types is specialized to be fast and Ca(2+)-dependent suggesting the involvement of other regulatory proteins specific for regulated exocytosis. Among these are the complexins, two closely related proteins that bind only to the assembled SNARE complex. We have investigated the function of complexin by analysis of single vesicle release events in adrenal chromaffin cells using carbon fiber amperometry. These cells express complexin II, and overexpression of this protein modified the kinetics of vesicle release events so that their time course was shortened. This effect depended on complexin interaction with the SNARE complex as introduction of a mutation of Arg-59, a residue that interacts with synaptobrevin in the SNARE complex, abolished its effects. The data are consistent with a function for complexin in stabilizing an intermediate of the SNARE complex to allow kiss-and run recycling of the exocytosed vesicle. PMID- 11929861 TI - A novel group of oleosins is present inside the pollen of Arabidopsis. AB - In plants, subcellular triacylglycerol granules in seeds (oil bodies) and floral tapetum (tapetosomes) are stabilized by amphipathic structural protein called oleosin. We hereby report a novel group of oleosins that is present inside the pollen of Arabidopsis thaliana. We have used the conserved sequence of oleosins to locate, via the DNA database, all 16 oleosin genes in the Arabidopsis genome. The oleosin genes can be divided into three groups according to their sequences and tissue-specific expressions, as probed by RNA blot hybridization and reverse transcriptase-PCR. The first group includes eight genes specifically expressed in the floret tapetum. The second group includes five genes specifically expressed in maturing seeds. The third, novel group includes three genes expressed in both maturing seeds and floral microspores, which will become pollen. Transgenic study using the promoter of one of these genes attached to a reporter gene has provided corroborative evidence for the specific expression of the gene in the microspores in the florets. One of the pollen oleosins can be identified by microsequencing and specific immunoblotting. Pollen oleosins synthesized by recombinant bacteria can collaborate with phospholipids in stabilizing reconstituted oil bodies. Thus, pollen has oleosins to stabilize the abundant subcellular oil bodies. Seed oil bodies and floret tapetosomes have been isolated from the miniature Arabidopsis plants, and the success indicates that the organelles can be subjected to future biochemical and genetic studies. PMID- 11929860 TI - Characterization of the murine hyaluronidase gene region reveals complex organization and cotranscription of Hyal1 with downstream genes, Fus2 and Hyal3. AB - Hyaluronidases are required for the breakdown of hyaluronan (HA), an abundant component of the extracellular matrix of vertebrate tissues. Multiple hyaluronidase genes have been identified, but the only clue to the function of their products has come from the identification of hyaluronidase 1 deficiency in a single patient with a mild clinical phenotype. As a first step in the generation of mice with hyaluronidase deficiency, we have used experimental and bioinformatic approaches to examine the organization of the mouse chromosome 9 region containing, in order, Hyal2, Hyal1, and Hyal3. This region was found to be complex, with Fus2 partially embedded in Hyal3, and Ifrd2 immediately downstream from Hyal3. The Hyal genes were all found to have four exons, and exons 2-4 exhibited the highest sequence conservation. Northern blot analysis demonstrated that the tissue expression profile for Hyal1 was similar in mice and humans, but a greater number of transcripts was detected in mouse tissues. Hyal3 was expressed more broadly in mice compared with humans and again exhibited additional transcripts. Reverse transcription-PCR demonstrated that some of the larger Hyal1 transcripts, seen on the Northern blot, were the result of cotranscription of Hyal1 with downstream genes, Fus2 or Hyal3. In vitro transcription/translation of one of the high abundance bicistronic transcripts produced Hyal 1, suggesting that Hyal 1 could be produced from all of the bicistronic transcripts. Characterization of the region including mouse Hyal1 and Hyal3 revealed complex organization and transcription that must be considered in the development and interpretation of mouse models involving genes in this region. PMID- 11929862 TI - In vitro evolution of recognition specificity mediated by SH3 domains reveals target recognition rules. AB - We have designed a repertoire of 10(7) different SH3 domains by grafting the residues that are represented in the binding surfaces of natural SH3 domains onto the scaffold of the human Abl-SH3 domain. This phage-displayed library was screened by affinity selection for SH3 domains that bind to the synthetic peptides, APTYPPPLPP and LSSRPLPTLPSP, which are peptide ligands for the human Abl or Src SH3 domains, respectively. By characterizing the isolates, we have observed that as few as two or three amino acid substitutions lead to dramatic changes in recognition specificity. We propose that the ability to shift recognition specificity with a small number of amino acid replacements is an important evolutionary characteristic of protein binding modules. Furthermore, we have used the information obtained by these in vitro evolution experiments to generate a scoring matrix that evaluates the probability that any SH3 domain binds to the peptide ligands for the Abl and Src SH3 domains. A table of predictions for the 28 SH3 domains of baker's yeast is presented. PMID- 11929863 TI - Activation of matrix metalloproteinase-2 by overexpression of manganese superoxide dismutase in human breast cancer MCF-7 cells involves reactive oxygen species. AB - Matrix metalloproteinases (MMPs) participate in cell migration and remodeling processes by affecting the extracellular matrix. MMP-2 is thought to be involved in cancer cell invasiveness. It has been proposed that the activity of MMP-2 can be modulated by intracellular reactive oxygen species (ROS)/reactive nitrogen species. We hypothesized that manganese superoxide dismutase (MnSOD) could mediate MMP-2 activity by changing the intracellular ROS level and that nitric oxide ((.)NO) may be involved in this process. Human breast cancer MCF-7 cells were stably transfected with plasmids containing MnSOD cDNA. A 2-30-fold increase of MnSOD protein and activity was observed in four clones. Our data demonstrated that overexpression of MnSOD stimulated the activation of MMP-2 with a corresponding elevation of ROS. A decrease in ROS by ebselen, a glutathione peroxidase mimetic, or by transduction of adenovirus containing human catalase or glutathione peroxidase cDNA abolished the effect of MnSOD on MMP-2 activation. Treatment of MCF-7 cells with antimycin A or rotenone increased intracellular ROS production and MMP-2 activation simultaneously. Our data also showed a suppression of endothelial nitric-oxide synthase expression that was accompanied by decreased (.)NO production in MnSOD-overexpressing cells. However, the changes in endothelial nitric-oxide synthase and (.)NO did not correlate with the MnSOD activity. Corresponding changes of MMP-2 activity after the addition of a NOS inhibitor (N(G)-amino-l-arginine) or a (.)NO donor ((Z)-1-[(2-aminoethyl)-N-(2 ammonioethyl)amino]diazen-1-ium-1,2-diolate) to the cells suggested the possibility that (.)NO may be involved in the MnSOD-mediated MMP-2 activation pathway. These results indicate that MnSOD induces MMP-2 activity by regulation of intracellular ROS and imply that signaling pathways involving (.)NO may also be involved in the MnSOD mediation of MMP-2 activity. PMID- 11929864 TI - Targeted gap junction protein constructs reveal connexin-specific differences in oligomerization. AB - To define further the mechanisms of gap junction protein (connexin (Cx)) oligomerization without pharmacologic disruption, we have examined the transport and assembly of connexin constructs containing C-terminal di-lysine-based endoplasmic reticulum (ER) (HKKSL) or ER-Golgi intermediate compartment (AKKFF) targeting sequences. By immunofluorescence microscopy, Cx43-HKKSL transiently transfected into HeLa cells showed a predominantly ER localization, although Cx43 AKKFF was localized to the perinuclear region of the cell. Sucrose gradient analysis of Triton X-100-solubilized connexins showed that either Cx43-HKKSL or Cx43-AKKFF expressed alone by HeLa cells was maintained as an apparent monomer. In contrast to Cx43-HKKSL, Cx32-HKKSL was maintained in the ER as stable hexamers, consistent with the notion that Cx32 and Cx43 oligomerization occur in distinct intracellular compartments. Furthermore, Cx43-HKKSL and Cx43-AKKFF inhibited trafficking of Cx43 and Cx46 to the plasma membrane. The inhibitory effect was because of the formation of mixed oligomers between Cx43-HKKSL or Cx43 AKKF and wild type Cx43 or Cx46. Taken together, these results suggest that Cx43 HKKSL and Cx43-AKKFF recirculate through compartments where oligomerization occurs and may be maintained as apparent monomers by a putative Cx43-specific quality control mechanism. PMID- 11929865 TI - Proteolysis of latent transforming growth factor-beta (TGF-beta )-binding protein 1 by osteoclasts. A cellular mechanism for release of TGF-beta from bone matrix. AB - The binding of growth factors to the extracellular matrix (ECM) may be a key pathway for regulation of their activity. We have shown that a major mechanism for storage of transforming growth factor-beta (TGF-beta) in bone ECM is via its association with latent TGF-beta-binding protein-1 (LTBP1). Although proteolytic cleavage of LTBP1 has been reported, it remains unclear whether this represents a physiological mechanism for release of matrix-bound TGF-beta. Here we examined the role of LTBP1 in cell-mediated release of TGF-beta from bone ECM. We first characterized the soluble and ECM-bound forms of latent TGF-beta produced by primary osteoblasts. Next, we examined release of ECM-bound TGF-beta by bone resorbing cells. Isolated avian osteoclasts and rabbit bone marrow-derived osteoclasts released bone matrix-bound TGF-beta via LTBP1 cleavage. 1,25 Dihydroxyvitamin D3 enhanced LTBP1 cleavage, resulting in release of 90% of the ECM-bound LTBP1. In contrast, osteoblasts failed to cleave LTBP1 or release TGF beta from bone ECM. Cleavage of LTBP1 by avian osteoclasts was inhibited by serine protease and metalloproteinase (MMP) inhibitors. Studies using purified proteases showed that plasmin, elastase, MMP2, and MMP9 were able to cleave LTBP1 to produce 125-165-kDa fragments. These studies identify LTBP1 as a novel substrate for MMPs and provide the first demonstration that LTBP1 proteolysis may be a physiological mechanism for release of TGF-beta from ECM-bound stores, potentially the first step in the pathway by which matrix-bound TGF-beta is rendered active. PMID- 11929866 TI - Mer receptor tyrosine kinase signaling: prevention of apoptosis and alteration of cytoskeletal architecture without stimulation or proliferation. AB - Mer is a member of the Axl/Mer/Tyro3 receptor tyrosine kinase family, a family whose physiological function is not well defined. We constructed a Mer chimera using the epidermal growth factor receptor (EGFR) extracellular and transmembrane domains and the Mer cytoplasmic domain. Stable transfection of the Mer chimera into interleukin 3 (IL-3)-dependent murine 32D cells resulted in ligand-activable surface receptor that tyrosine autophosphorylated, stimulated intracellular signaling, and dramatically reduced apoptosis initiated by IL-3 withdrawal. However, unlike multiple other ectopically expressed receptor tyrosine kinases including full-length EGFR or an EGFR/Axl chimera, the Mer chimera did not stimulate proliferation. Moreover, and in contrast to EGFR, Mer chimera activation induced adherence and cell flattening in the normally suspension growing 32D cells. The Mer chimera signal also blocked IL-3-dependent proliferation leading to G(1)/S arrest, dephosphorylation of retinoblastoma protein, and elongation of cellular processes. Unlike other agonists that lead to a slow (4-8 days) ligand-dependent differentiation of 32D cells, the combined Mer and IL-3 signal resulted in differentiated morphology and growth cessation in the first 24 h. Thus the Mer chimera blocks apoptosis without stimulating growth and produces cytoskeletal alterations; this outcome is clearly separable from the proliferative signal produced by most receptor tyrosine kinases. PMID- 11929867 TI - Phosphorylation of neuroglycan C, a brain-specific transmembrane chondroitin sulfate proteoglycan, and its localization in the lipid rafts. AB - Neuroglycan C (NGC) is a brain-specific transmembrane chondroitin sulfate proteoglycan. In the present study, we examined whether NGC could be phosphorylated in neural cells. On metabolic labeling of cultured cerebral cortical cells from the rat fetus with (32)P(i), serine residues in NGC were radiolabeled. Some NGC became detectable in the raft fraction from the rat cerebrum, a signaling microdomain of the plasma membrane, with cerebral development. NGC from the non-raft fraction, not the raft fraction, could be phosphorylated by an in vitro kinase reaction. The phosphorylation of NGC was inhibited by adding to the reaction mixture a recombinant peptide representing the ectodomain of NGC, but not by adding a peptide representing its cytoplasmic domain. NGC could be labeled by an in vitro kinase reaction using [gamma (32)P]GTP as well as [gamma-(32)P]ATP, and this kinase activity was partially inhibited by 5,6-dichloro-1-beta-d-ribofuranosylbenzimidazole, a selective inhibitor of casein kinase II. In addition to the intracellular phosphorylation, NGC was also phosphorylated at the cell surface by an ectoprotein kinase. This is the first report to demonstrate that NGC can be phosphorylated both intracellularly and pericellularly, and our findings suggest that a kinase with a specificity similar to that of casein kinase II is responsible for the NGC ectodomain phosphorylation. PMID- 11929868 TI - Distinct regulatory effects of the Na,K-ATPase gamma subunit. AB - The two variants of the gamma subunit of the rat renal sodium pump, gamma(a) and gamma(b), have similar effects on the Na,K-ATPase. Both increase the affinity for ATP due to a shift in the enzyme's E(1) <--> E(2) conformational equilibrium toward E(1). In addition, both increase K(+) antagonism of cytoplasmic Na(+) activation. To gain insight into the structural basis for these distinct effects, extramembranous N-terminal and C-terminal mutants of gamma were expressed in rat alpha1-transfected HeLa cells. At the N terminus, the variant-distinct region was deleted (gammaNDelta7) or replaced by alanine residues (gammaN7A). At the C terminus, four (gamma(a)CDelta4) or ten (gamma(a)CDelta10) residues were deleted. None of these mutations abrogates the K(+)/Na(+) antagonism as evidenced in a similar increase in K'(Na) seen at high (100 mm) K(+) concentration. In contrast, the C-terminal as well as N-terminal deletions (gammaNDelta7, gamma(a)CDelta4, and gamma(a)CDelta10) abolished the decrease in K'(ATP) seen with wild-type gamma(a) or gamma(b). It is concluded that different regions of the gamma chain mediate the distinct functional effects of gamma, and the effects can be long range. In the transmembrane region, the impact of G41R replacement was analyzed since this mutation is associated with autosomal dominant renal Mg(2+)-wasting in man (Meij, I. C., Koenderink, J. B., van Bokhoven, H., Assink, K. F. H., Groenestege, W. T., de Pont, J. J. H. H. M., Bindels, R. J. M., Monnens, L. A. H., Van den Heuvel, L. P. W. J., and Knoers, N. V. A. M. (2000) Nat. Genet. 26, 265-266). The results show that Gly-41 --> Arg prevents trafficking of gamma but not alphabeta pumps to the cell surface and abrogates functional effects of gamma on alphabeta pumps. These findings underscore a potentially important role of gamma in affecting solute transport, in this instance Mg(2+) reabsorption, consequent to its primary effect on the sodium pump. PMID- 11929869 TI - A bicyclic autotrophic CO2 fixation pathway in Chloroflexus aurantiacus. AB - Phototrophic CO(2) assimilation by the primitive, green eubacterium Chloroflexus aurantiacus has been shown earlier to proceed in a cyclic mode via 3 hydroxypropionate, propionyl-CoA, succinyl-CoA, and malyl-CoA. The metabolic cycle could be closed by cleavage of malyl-CoA affording glyoxylate (the primary CO(2) fixation product) with regeneration of acetyl-CoA serving as the starter unit of the cycle. The pathway of glyoxylate assimilation to form gluconeogenic precursors has not been elucidated to date. We could now show that the incubation of cell extract with a mixture of glyoxylate and [1,2,3-(13)C(3)]propionyl-CoA afforded erythro-beta-[1,2,2'-(13)C(3)]methylmalate and [1,2,2' (13)C(3)]citramalate. Similar experiments using a partially purified protein fraction afforded erythro-beta-[1,2,2'-(13)C(3)]methylmalyl-CoA and [1,2,2' (13)C(3)]mesaconyl-CoA. Cell extracts of C. aurantiacus were also shown to catalyze the conversion of citramalate into pyruvate and acetyl-CoA in a succinyl CoA-dependent reaction. The data suggest that glyoxylate obtained by the cleavage of malyl-CoA can be utilized by condensation with propionyl-CoA affording erythro beta-methylmalyl-CoA, which is converted to acetyl-CoA and pyruvate. This reaction sequence regenerates acetyl-CoA, which serves as the precursor of propionyl-CoA in the 3-hydroxypropionate cycle. Autotrophic CO(2) fixation proceeds by combination of the 3-hydroxypropionate cycle with the methylmalyl-CoA cycle. The net product of that bicyclic autotrophic CO(2) fixation pathway is pyruvate serving as an universal building block for anabolic reactions. PMID- 11929870 TI - Multiple mechanisms linked to platelet activation result in lysophosphatidic acid and sphingosine 1-phosphate generation in blood. AB - Lysophosphatidic acid (LPA) and sphingosine 1-phosphate (Sph1P) production was examined in vitro under conditions that simulated blood clotting. Several approaches were utilized to elucidate the metabolic pathways. 1) Platelet phospholipids were labeled using [32P]orthophosphate, and the production of [32P]Sph1P and LPA was examined. Thrombin stimulation of platelets resulted in rapid secretion of Sph1P stored within the platelet. In contrast, LPA was neither stored within nor secreted from platelets. Nonetheless, extracellular levels of LPA gradually increased following stimulation. 2) Stable-isotope dilution mass spectrometry was used to quantify the molecular species of LPA generated from platelets in vitro. Only 10% of the LPA generated following thrombin stimulation was associated with platelets, the remaining 90% was contained within the extracellular medium. The acyl composition of LPA produced by platelets differed depending on the presence or absence of plasma in the incubation. 3) The fate of exogenously added fluorescent phospholipid analogs was determined. Incubation of [(7-nitro-2-1,3-benzoxadiazol-4-yl)amino]dodecanoyl-(NBD)-labeled phosphatidylcholine, phosphatidylethanolamine, and phosphatidylserine with the supernatant fractions from thrombin-stimulated platelets yielded no LPA production. However, these lipids were converted to the corresponding lysolipids by released PLA1 and PLA2 activities. When incubated with plasma or serum the NBD labeled lysophospholipids were readily converted to LPA. Inhibitors of lysophospholipase D and the biological activity of LPA were detected in plasma. These results suggest that the bulk of LPA produced through platelet activation results from the sequential cleavage of phospholipids to lysophospholipids by released phospholipases A1 and A2 and then to LPA by plasma lysophospholipase D. PMID- 11929872 TI - Functional characterization of Drosophila melanogaster peptide O xylosyltransferase, the key enzyme for proteoglycan chain initiation and member of the core 2/I N-acetylglucosaminyltransferase family. AB - Chondroitin and heparan sulfates are essential players in animal development and are synthesized by a series of glycosyltransferases, the first of which is UDP alpha-D-xylose:proteoglycan core protein beta-D-xylosyltransferase (EC ). In the present study, a Drosophila melanogaster gene (CG17771), previously designated as a homologue of core 2 and I beta1,6-N-acetylglucosaminyltransferases, was shown to encode an active peptide O-xylosyltransferase. A novel coupled assay using matrix-assisted laser desorption ionization time-of-flight mass spectrometry demonstrated transfer of xylose to the peptide DDDSIEGSGGR. Analysis of sequences of various peptide O-xylosyltransferase and beta1,6-N acetylglucosaminyltransferase sequences indicates that they are members of a large multifunctional protein family with a range of roles in beta-glycosylation of either peptide or glycan substrates. Because in contrast to mammals, there is only one fly peptide O-xylosyltransferase gene, it is anticipated that, given the key roles of proteoglycans, the hereby designated oxt gene is essential for viability. PMID- 11929871 TI - Bcl-2 family member Bfl-1/A1 sequesters truncated bid to inhibit is collaboration with pro-apoptotic Bak or Bax. AB - Following caspase-8 mediated cleavage, a carboxyl-terminal fragment of the BH3 domain-only Bcl-2 family member Bid transmits the apoptotic signal from death receptors to mitochondria. In a screen for possible regulators of Bid, we defined Bfl-1/A1 as a potent Bid interacting protein. Bfl-1 is an anti-apoptotic Bcl-2 family member, whose preferential expression in hematopoietic cells and endothelium is controlled by inflammatory stimuli. Its mechanism of action is unknown. We find that Bfl-1 associates with both full-length Bid and truncated (t)Bid, via the Bid BH3 domain. Cellular expression of Bfl-1 confers protection against CD95- and Trail receptor-induced cytochrome c release. In vitro assays, using purified mitochondria and recombinant proteins, demonstrate that Bfl-1 binds full-length Bid, but does not interfere with its processing by caspase-8, or with its mitochondrial association. Confocal microscopy supports that Bfl-1, which at least in part constitutively localizes to mitochondria, does not impede tBid translocation. However, Bfl-1 remains tightly and selectively bound to tBid and blocks collaboration between tBid and Bax or Bak in the plane of the mitochondrial membrane, thereby preventing mitochondrial apoptotic activation. Lack of demonstrable interaction between Bfl-1 and Bak or Bax in the mitochondrial membrane suggests that Bfl-1 generally prevents the formation of a pro-apoptotic complex by sequestering BH3 domain-only proteins. PMID- 11929873 TI - Class II histone deacetylases are directly recruited by BCL6 transcriptional repressor. AB - BCL6 is a member of the POZ/zinc finger (POK) family involved in survival and/or differentiation of a number of cell types and in B cell lymphoma upon chromosomal alteration. Transcriptional repression by BCL6 is thought to be achieved in part by recruiting a repressor complex containing two class I histone deacetylases (HDACs). In this study we investigated whether BCL6 could also target members of class II HDACs. Our results indicate that three related class II deacetylases, HDAC4, HDAC5, and HDAC7 can associate with BCL6 in vivo and in vitro. Using electron microscopy, we found that endogenous BCL6 and class II HDACs partially co-localize in the nucleus. Overexpression experiments showed that BCL6 and HDAC4, -5, or -7 are intermingled onto common nuclear substructures and form stable complexes. A highly conserved domain in the N-terminal region of HDAC5 and HDAC7 as well as the zinc finger region of BCL6 were found necessary for the complex formation in vivo and in vitro. Moreover, our data point to the zinc finger region of BCL6 as a multifunctional domain which, beside its known capacity to bind DNA, is involved in the nuclear targeting of the protein and in the recruitment of the class II HDACs, and hence constitutes an autonomous repressor domain. Since PLZF, a BCL6 relative, could also interact with HDAC4, 5, and 7, we suggest that class II HDACs are largely involved in the control of the POK transcription factors activity. PMID- 11929874 TI - A functional role for the B56 alpha-subunit of protein phosphatase 2A in ceramide mediated regulation of Bcl2 phosphorylation status and function. AB - Recently it has been shown that the potent apoptotic agent ceramide activates a mitochondrial protein phosphatase 2A (PP2A) and promotes dephosphorylation of the anti-apoptotic molecule Bcl2 (Ruvolo, P. P., Deng, X., Ito, T., Carr, B. K., and May, W. S. (1999) J. Biol. Chem. 274, 20296-20300). In cells expressing Bcl2, dephosphorylation of Bcl2 appears to be required for ceramide-induced cell death because treatment of cells with low doses of the PP2A inhibitor okadaic acid blocks Bcl2 dephosphorylation and promotes cell survival. Furthermore, the non phosphorylatable (i.e. PP2A-resistant) gain-of-function S70E mutant Bcl2 can protect cells from ceramide-induced apoptosis. These findings support a model whereby Bcl2 function is regulated by PP2A. PP2A is a heterotrimer that contains a catalytic C-subunit, a structural A-subunit, and a regulatory B-subunit. The A- and C-subunits are fairly conserved and ubiquitously expressed, and they form the catalytic complex of the phosphatase. In contrast, there are at least three families of diverse B-subunit molecules that vary in expression temporally and by tissue type. It is hypothesized that ceramide regulates PP2A via the B-subunit. Thus, understanding the mechanism of how PP2A regulates Bcl2 phosphorylation status and how ceramide might regulate this process requires identification of the regulatory B-subunit of PP2A that comprises the Bcl2 phosphatase. Results indicate that the B56 alpha-subunit is a candidate regulatory subunit of the physiologic Bcl2 phosphatase since (a) B56 alpha associates with Bcl2 as evidenced by pull-down experiments, (b) B56 alpha co-localizes with Bcl2 in mitochondrial membranes, (c) ceramide promotes translocation of B56 alpha to mitochondrial membranes, and (d) overexpression of B56 alpha promotes mitochondrial PP2A activity and Bcl2 dephosphorylation and potentiates cell killing with ceramide. These findings suggest a role for B56 alpha in regulating the Bcl2 phosphatase. PMID- 11929876 TI - Dynamic regulation of LFA-1 activation and neutrophil arrest on intercellular adhesion molecule 1 (ICAM-1) in shear flow. AB - Neutrophil recruitment during acute inflammation is triggered by G-protein-linked chemotactic receptors that in turn activate beta(2) integrin (CD18), deemed a critical step in facilitating cell capture and arrest under the shear force of blood flow. A conformational switch in the I domain allosteric site (IDAS) and in CD18 regulates LFA-1 affinity for endothelial ligands including intercellular adhesion molecule 1 (ICAM-1). We examined the dynamics of CD18 activation in terms of the efficiency of neutrophil capture of ICAM-1, and we correlated this with the membrane topography of 327C, an antibody that recognizes the active conformation of CD18 I-like domain. Adhesion increased in direct proportion to chemotactic stimulus rising 7-fold over a log range of interleukin-8 (IL-8). A threshold dose of approximately 75 pm IL-8, corresponding to ligation of only approximately 10-100 receptors, was sufficient to activate approximately 20,000 CD18 and a rapid boost in the capture efficiency on ICAM-1. This was accompanied by a rapid redistribution of active LFA-1, but not Mac-1, into membrane patches, a necessary component for optimum adhesion efficiency. Shear-resistant arrest on a monolayer of ICAM-1 was reversed within minutes of chemotactic stimulation correlating with a shift from high to low affinity CD18 and dispersal of patches of active CD18. Mobility of active CD18 into high avidity patches was dependent on phosphatidylinositol 3-kinase activity and not F-actin polymerization. The data reveal that the number of chemotactic receptors bound and the topography and lifetime of high affinity LFA-1 tightly regulate the efficiency of neutrophil capture on ICAM-1. PMID- 11929877 TI - Sphingomyelin and cholesterol promote HIV-1 gp41 pretransmembrane sequence surface aggregation and membrane restructuring. AB - The interfacial sequence DKWASLWNWFNITNWLWYIK, preceding the transmembrane anchor of gp41 glycoprotein subunit, has been shown to be essential for fusion activity and incorporation into virions. HIV(c), a peptide representing this region, formed lytic pores in liposomes composed of the main lipids occurring in the human immunodeficiency virus, type 1 (HIV-1), envelope, i.e. 1-palmitoyl-2 oleoylphosphatidylcholine (POPC):sphingomyelin (SPM):cholesterol (Chol) (1:1:1 mole ratio), at low (>1:10,000) peptide-to-lipid mole ratio, and promoted the mixing of vesicular lipids at >1:1000 peptide-to-lipid mole ratios. Inclusion of SPM or Chol in POPC membranes had different effects. Whereas SPM sustained pore formation, Chol promoted fusion activity. Even if partitioning into membranes was not affected in the absence of both SPM and Chol, HIV(c) had virtually no effect on POPC vesicles. Conditions described to disturb occurrence of lateral separation of phases in these systems reproduced the high peptide-dose requirements for leakage as found in pure POPC vesicles and inhibited fusion. Surface aggregation assays using rhodamine-labeled peptides demonstrated that SPM and Chol promoted HIV(c) self-aggregation in membranes. Employing head-group fluorescent phospholipid analogs in planar supported lipid layers, we were able to discern HIV(c) clusters associated to ordered domains. Our results support the notion that the pretransmembrane sequence may participate in the clustering of gp41 monomers within the HIV-1 envelope, and in bilayer architecture destabilization at the loci of fusion. PMID- 11929878 TI - Sec34 is implicated in traffic from the endoplasmic reticulum to the Golgi and exists in a complex with GTC-90 and ldlBp. AB - Sec34p/Grd20p has been implicated in endoplasmic reticulum (ER)-to-Golgi transport and/or post-Golgi trafficking events and exists in a protein complex consisting of at least eight subunits in yeast. Although the mammalian counterpart (Sec34) of Sec34p has been molecularly identified, its role and interacting partners remain undefined. In this study, we have prepared antibodies specifically against the recombinant N-terminal fragment of Sec34 that recognize a polypeptide of about 93 kDa and label the Golgi apparatus. In a well characterized semi-intact cell assay that reconstitutes transport of the envelope glycoprotein (VSVG) of vesicular stomatitis virus from the ER to the Golgi apparatus, anti-Sec34 antibodies inhibited the transport in a dose-dependent manner. The inhibition by anti-Sec34 antibodies could be neutralized by a noninhibitory amount of the antigen. Large-scale immunoprecipitation of rat liver cytosol with immobilized anti-Sec34 antibodies has co-immunoprecipitated GTC-90 and ldlBp, two peripheral Golgi proteins previously shown to exist in separate protein complexes. Two mammalian homologues (Dor1 and Cod1) of the yeast Sec34 complex were similarly recovered in the Sec34 immunoprecipitates. When expressed in transfected cells, epitope-tagged ldlCp and Cod2 were co-immunoprecipitated with anti-Sec34 antibodies with efficiencies comparable to that observed for tagged ldlBp, Dor1, and Cod1. Direct interactions of Sec34 with ldlBp and ldlCp were further demonstrated in vitro. These results suggest that Sec34, GTC-90, and ldlBp/ldlCp are part of the same protein complex(es) that regulates diverse aspects of Golgi function, including transport from the ER to the Golgi apparatus. PMID- 11929879 TI - Proliferating cell nuclear antigen associates with histone deacetylase activity, integrating DNA replication and chromatin modification. AB - Faithful inheritance of the chromatin structure is essential for maintaining the gene expression integrity of a cell. Histone modification by acetylation and deacetylation is a critical control of chromatin structure. In this study, we test the hypothesis that histone deacetylase 1 (HDAC1) is physically associated with a basic component of the DNA replication machinery as a mechanism of coordinating histone deacetylation and DNA synthesis. Proliferating cell nuclear antigen (PCNA) is a sliding clamp that serves as a loading platform for many proteins involved in DNA replication and DNA repair. We show that PCNA interacts with HDAC1 in human cells and in vitro and that a considerable fraction of PCNA and HDAC1 colocalize in the cell nucleus. PCNA associates with histone deacetylase activity that is completely abolished in the presence of the HDAC inhibitor trichostatin A. Trichostatin A treatment arrests cells at the G(2)-M phase of the cell cycle, which is consistent with the hypothesis that the proper formation of the chromatin after DNA replication may be important in signaling the progression through the cell cycle. Our results strengthen the role of PCNA as a factor coordinating DNA replication and epigenetic inheritance. PMID- 11929880 TI - Protein phosphatase 2A holoenzyme assembly: identification of contacts between B family regulatory and scaffolding A subunits. AB - Protein serine/threonine phosphatase (PP) 2A is a ubiquitous enzyme with pleiotropic functions. Trimeric PP2A consists of a structural A subunit, a catalytic C subunit, and a variable regulatory subunit. Variable subunits (B, B', and B" families) dictate PP2A substrate specificity and subcellular localization. B-family subunits contain seven WD repeats predicted to fold into a beta propeller structure. We carried out mutagenesis of Bgamma to identify domains important for association with A and C subunits in vivo. Several internal deletions in Bgamma abolished coimmunoprecipitation of A and C subunits expressed in COS-M6 cells. In contrast, small N- and C-terminal Bgamma deletions had no effect on incorporation into the PP2A heterotrimer. Thus, holoenzyme association of B-family subunits requires multiple, precisely aligned contacts within a core beta-propeller domain. Charge-reversal mutagenesis of Bgamma identified a cluster of conserved critical residues in Bgamma WD repeats 3 and 4. Acidic substitution of paired basic residues in Bgamma (RR165EE) abolished association with wild-type A and C subunits, while fostering incorporation of Bgamma into a PP2A heterotrimer containing an A subunit with an opposite charge-reversal mutation (EE100RR). Thus, binding of A and B subunits requires electrostatic interactions between conserved pairs of glutamates and arginines. By expressing complementary charge-reversal mutants in neuronal PC6-3 cells, we further show that holoenzyme incorporation protects Bgamma from rapid degradation by the ubiquitin/proteasome pathway. PMID- 11929881 TI - Acute stimulation of Na/K pump by cardiac glycosides in the nanomolar range. PMID- 11929882 TI - Isoform-specific stimulation of cardiac Na/K pumps by nanomolar concentrations of glycosides. AB - It is well-known that micromolar to millimolar concentrations of cardiac glycosides inhibit Na/K pump activity, however, some early reports suggested nanomolar concentrations of these glycosides stimulate activity. These early reports were based on indirect measurements in multicellular preparations, hence, there was some uncertainty whether ion accumulation/depletion rather than pump stimulation caused the observations. Here, we utilize the whole-cell patch-clamp technique on isolated cardiac myocytes to directly measure Na/K pump current (I(P)) in conditions that minimize the possibility of ion accumulation/depletion causing the observed effects. In guinea pig ventricular myocytes, nanomolar concentrations of dihydro-ouabain (DHO) caused an outward current that appeared to be due to stimulation of I(P) because of the following: (1) it was absent in 0 mM [K(+)](o), as was I(P); (2) it was absent in 0 mM [Na(+)](i), as was I(P); (3) at reduced [Na(+)](i), the outward current was reduced in proportion to the reduction in I(P); (4) it was eliminated by intracellular vanadate, as was I(P). Our previous work suggested guinea pig ventricular myocytes coexpress the alpha(1)- and alpha(2)-isoforms of the Na/K pumps. The stimulation of I(P) appears to be through stimulation of the high glycoside affinity alpha(2)-isoform and not the alpha(1)-isoform because of the following: (1) regulatory signals that specifically increased activity of the alpha(2)-isoform increased the amplitude of the stimulation; (2) regulatory signals that specifically altered the activity of the alpha(1)-isoform did not affect the stimulation; (3) changes in [K(+)](o) that affected activity of the alpha(1)-isoform, but not the alpha(2) isoform, did not affect the stimulation; (4) myocytes from one group of guinea pigs expressed the alpha(1)-isoform but not the alpha(2)-isoform, and these myocytes did not show the stimulation. At 10 nM DHO, total I(P) increased by 35 +/- 10% (mean +/- SD, n = 18). If one accepts the hypothesis that this increase is due to stimulation of just the alpha(2)-isoform, then activity of the alpha(2) isoform increased by 107 +/- 30%. In the guinea pig myocytes, nanomolar ouabain as well as DHO stimulated the alpha(2)-isoform, but both the stimulatory and inhibitory concentrations of ouabain were approximately 10-fold lower than those for DHO. Stimulation of I(P) by nanomolar DHO was observed in canine atrial and ventricular myocytes, which express the alpha(1)- and alpha(3)-isoforms of the Na/K pumps, suggesting the other high glycoside affinity isoform (the alpha(3) isoform) also was stimulated by nanomolar concentrations of DHO. Human atrial and ventricular myocytes express all three isoforms, but isoform affinity for glycosides is too similar to separate their activity. Nevertheless, nanomolar DHO caused a stimulation of I(P) that was very similar to that seen in other species. Thus, in all species studied, nanomolar DHO caused stimulation of I(P), and where the contributions of the high glycoside affinity alpha(2)- and alpha(3)-isoforms could be separated from that of the alpha(1)-isoform, it was only the high glycoside affinity isoform that was stimulated. These observations support early reports that nanomolar concentrations of glycosides stimulate Na/K pump activity, and suggest a novel mechanism of isoform-specific regulation of I(P) in heart by nanomolar concentrations of endogenous ouabain-like molecules. PMID- 11929883 TI - Interactions among toxins that inhibit N-type and P-type calcium channels. AB - A number of peptide toxins from venoms of spiders and cone snails are high affinity ligands for voltage-gated calcium channels and are useful tools for studying calcium channel function and structure. Using whole-cell recordings from rat sympathetic ganglion and cerebellar Purkinje neurons, we studied toxins that target neuronal N-type (Ca(V)2.2) and P-type (Ca(V)2.1) calcium channels. We asked whether different toxins targeting the same channels bind to the same or different sites on the channel. Five toxins (omega-conotoxin-GVIA, omega conotoxin MVIIC, omega-agatoxin-IIIA, omega-grammotoxin-SIA, and omega-agatoxin IVA) were applied in pairwise combinations to either N- or P-type channels. Differences in the characteristics of inhibition, including voltage dependence, reversal kinetics, and fractional inhibition of current, were used to detect additive or mutually occlusive effects of toxins. Results suggest at least two distinct toxin binding sites on the N-type channel and three on the P-type channel. On N-type channels, results are consistent with blockade of the channel pore by omega-CgTx-GVIA, omega-Aga-IIIA, and omega-CTx-MVIIC, whereas grammotoxin likely binds to a separate region coupled to channel gating. omega-Aga-IIIA produces partial channel block by decreasing single-channel conductance. On P type channels, omega-CTx-MVIIC and omega-Aga-IIIA both likely bind near the mouth of the pore. omega-Aga-IVA and grammotoxin each bind to distinct regions associated with channel gating that do not overlap with the binding region of pore blockers. For both N- and P-type channels, omega-CTx-MVIIC binding produces complete channel block, but is prevented by previous partial channel block by omega-Aga-IIIA, suggesting that omega-CTx-MVIIC binds closer to the external mouth of the pore than does omega-Aga-IIIA. PMID- 11929884 TI - The mechanism of ciliary stimulation by acetylcholine: roles of calcium, PKA, and PKG. AB - Stimulation of ciliary cells through muscarinic receptors leads to a strong biphasic enhancement of ciliary beat frequency (CBF). The main goal of this work is to delineate the chain of molecular events that lead to the enhancement of CBF induced by acetylcholine (ACh). Here we show that the Ca(2+), cGMP, and cAMP signaling pathways are intimately interconnected in the process of cholinergic ciliary stimulation. ACh induces profound time-dependent increase in cGMP and cAMP concentrations mediated by the calcium-calmodulin complex. The initial strong CBF enhancement in response to ACh is mainly governed by PKG and elevated calcium. The second phase of CBF enhancement induced by ACh, a stable moderately elevated CBF, is mainly regulated by PKA in a Ca(2+)-independent manner. Inhibition of either guanylate cyclase or of PKG partially attenuates the response to ACh of [Ca(2+)](i), but completely abolishes the response of CBF. Inhibition of PKA moderately attenuates and significantly shortens the responses to ACh of both [Ca(2+)](i) and CBF. In addition, PKA facilitates the elevation in [Ca(2+)](i) and cGMP levels induced by ACh, whereas an unimpeded PKG activity is essential for CBF enhancement mediated by either Ca(2+) or PKA. PMID- 11929885 TI - Voltage-dependence of ion permeation in cyclic GMP-gated ion channels is optimized for cell function in rod and cone photoreceptors. AB - The kinetics of the photocurrent in both rod and cone retinal photoreceptors are independent of membrane voltage over the physiological range (-30 to -65 mV). This is surprising since the photocurrent time course is regulated by the influx of Ca(2+) through cGMP-gated ion channels (CNG) and the force driving this flux changes with membrane voltage. To understand this paradigm, we measured Pf, the fraction of the cyclic nucleotide-gated current specifically carried by Ca(2+) in intact, isolated photoreceptors. To measure Pf we activated CNG channels by suddenly increasing free 8-Br-cGMP in the cytoplasm of rods or cones loaded with a caged ester of the cyclic nucleotide. Simultaneous with the uncaging flash, we measured the cyclic nucleotide-dependent changes in membrane current and fluorescence of the Ca(2+) binding dye, Fura-2, also loaded into the cells. We determined Pf under physiological solutions at various holding membrane voltages between -65 and -25 mV. Pf is larger in cones than in rods, but in both photoreceptor types its value is independent of membrane voltage over the range tested. This biophysical feature of the CNG channels offers a functional advantage since it insures that the kinetics of the phototransduction current are controlled by light, and not by membrane voltage. To explain our observation, we developed a rate theory model of ion permeation through CNG channels that assumes the existence of two ion binding sites within the permeation pore. To assign values to the kinetic rates in the model, we measured experimental I-V curves in membrane patches of rods and cones over the voltage range -90 to 90 mV in the presence of simple biionic solutions at different concentrations. We optimized the fit between simulated and experimental data. Model simulations describe well experimental photocurrents measured under physiological solutions in intact cones and are consistent with the voltage-independence of Pf, a feature that is optimized for the function of the channel in photoreceptors. PMID- 11929888 TI - Desensitization of group I metabotropic glutamate receptors in rat sympathetic neurons. AB - Desensitization of heterologously expressed metabotropic glutamate receptor 5a (mGluR5a) was examined in rat sympathetic neurons. Calcium currents in cells expressing mGluR5a exhibited substantial inhibition in response to glutamate exposure. In the continued presence of glutamate, inhibition attenuated rapidly over the course of about a minute. Desensitization was eliminated when a nonhydrolyzable ATP analogue was substituted for ATP in the pipette solution, suggesting that desensitization was mediated by a phosphorylation event. Next, pharmacological agents were used to investigate the nature of the kinase involved in desensitization. Desensitization was sensitive to the nonspecific kinase inhibitor, staurosporine, but not H-7, another nonspecific kinase inhibitor. Inhibitors of myosin light chain kinase and calmodulin-dependent kinase were without effect on desensitization. However, desensitization was sensitive to the protein kinase C inhibitor bisindolymaleimide. In contrast, Go6976, a selective inhibitor of conventional protein kinase C isoforms, was without effect. In addition, desensitization persisted in the presence of 10 mM intracellular bis-(o aminophenoxy)-N,N,N',N'-tetraacetic acid, a fast Ca(2+) chelator. Finally, overexpression of wild-type calmodulin, which can bind mGluR5 and inhibit phosphorylation, did not alter mGluR desensitization. Two Ca(2+)-binding deficient calmodulin mutants were also without effect. These data indicate a role for nonconventional protein kinase C isoforms as a mediator of mGluR5 desensitization and that the phosphorylation of mGluR5a that competes with calmodulin binding does not mediate desensitization. PMID- 11929887 TI - Calcium-induced transitions between the spontaneous miniature outward and the transient outward currents in retinal amacrine cells. AB - Spontaneous miniature outward currents (SMOCs) occur in a subset of retinal amacrine cells at membrane potentials between -60 and -40 mV. At more depolarized potentials, a transient outward current (I(to)) appears and SMOCs disappear. Both SMOCs and the I(to) are K(+) currents carried by BK channels. They both arise from Ca(2+) influx through high voltage-activated (HVA) Ca(2+) channels, which stimulates release of internal Ca(2+) from caffeine- and ryanodine-sensitive stores. An increase in Ca(2+) influx resulted in an increase in SMOC frequency, but also led to a decline in SMOC mean amplitude. This reduction showed a temporal dependence: the effect being greater in the latter part of a voltage step. Thus, Ca(2+) influx, although required to generate SMOCs, also produced a negative modulation of their amplitudes. Increasing Ca(2+) influx also led to a decline in the first latency to SMOC occurrence. A combination of these effects resulted in the disappearance of SMOCs, along with the concomitant appearance of the I(to) at high levels of Ca(2+) influx. Therefore, low levels of Ca(2+) influx, arising from low levels of activation of the HVA Ca(2+) channels, produce randomly occurring SMOCs within the range of -60 to -40 mV. Further depolarization leads to greater activation of the HVA Ca(2+) channels, larger Ca(2+) influx, and the disappearance of discontinuous SMOCs, along with the appearance of the I(to). Based on their characteristics, SMOCs in retinal neurons may function as synaptic noise suppressors at quiescent glutamatergic synapses. PMID- 11929886 TI - Mechanism of generation of spontaneous miniature outward currents (SMOCs) in retinal amacrine cells. AB - A subtype of retinal amacrine cells displayed a distinctive array of K(+) currents. Spontaneous miniature outward currents (SMOCs) were observed in the narrow voltage range of -60 to -40 mV. Depolarizations above approximately -40 mV were associated with the disappearance of SMOCs and the appearance of transient (I(to)) and sustained (I(so)) outward K(+) currents. I(to) appeared at about -40 mV and its apparent magnitude was biphasic with voltage, whereas I(so) appeared near -30 mV and increased linearly. SMOCs, I(to), and a component of I(so) were Ca(2+) dependent. SMOCs were spike shaped, occurred randomly, and had decay times appreciably longer than the time to peak. In the presence of cadmium or cobalt, SMOCs with pharmacologic properties identical to those seen in normal Ringer's could be generated at voltages of -20 mV and above. Their mean amplitude was Nernstian with respect to [K(+)](ext) and they were blocked by tetraethylammonium. SMOCs were inhibited by iberiotoxin, were insensitive to apamin, and eliminated by nominally Ca(2+)-free solutions, indicative of BK-type Ca(2+)-activated K(+) currents. Dihydropyridine Ca(2+) channel antagonists and agonists decreased and increased SMOC frequencies, respectively. Ca(2+) permeation through the kainic acid receptor had no effect. Blockade of organelle Ca(2+) channels by ryanodine, or intracellular Ca(2+) store depletion with caffeine, eradicated SMOCs. Internal Ca(2+) chelation with 10 mM BAPTA eliminated SMOCs, whereas 10 mM EGTA had no effect. These results suggest a mechanism whereby Ca(2+) influx through L-type Ca(2+) channels and its subsequent amplification by Ca(2+)-induced Ca(2+) release via the ryanodine receptor leads to a localized elevation of internal Ca(2+). This amplified Ca(2+) signal in turn activates BK channels in a discontinuous fashion, resulting in randomly occurring SMOCs. PMID- 11929890 TI - Encoding of compressive stress during indentation by slowly adapting type I mechanoreceptors in rat hairy skin. AB - The mechanical state encoded by slowly adapting type 1 mechanoreceptors (SAI) during indentation was examined using an isolated preparation in a rat model. Skin and its intact innervation were harvested from the medial thigh of the rat hindlimb and placed in a dish, with the corium side down, containing synthetic interstitial fluid. The margins of the skin were coupled to an apparatus that could stretch and apply compression to the skin. Using a standard teased nerve preparation, the neural responses of single SAIs were identified. SAIs were stimulated, using controlled compressive stress while simultaneously measuring displacement, by compressing the skin between indenters (flat cylinders) of different diameters and a hard platform. SAIs were subcategorized according to whether their neural response saturated above or below 10 kPa compressive stress (SAI-H or SAI-L, respectively). Linear regression was used to evaluate the relationships between neuron response and stress and force and displacement. For all SAIs, the mean neural response was significantly and substantially more highly correlated with compressive stress than force or displacement. For the SAI L subcategory, the mean correlation coefficient was significantly and substantially greater for stress than for force but not significantly different for displacement. The data from this study support the hypothesis that SAI mechanoreceptors stimulated by indentation encode compressive stress rather than force, displacement, or strain. PMID- 11929889 TI - Role of eye, head, and shoulder geometry in the planning of accurate arm movements. AB - Eye-hand coordination requires the brain to integrate visual information with the continuous changes in eye, head, and arm positions. This is a geometrically complex process because the eyes, head, and shoulder have different centers of rotation. As a result, head rotation causes the eye to translate with respect to the shoulder. The present study examines the consequences of this geometry for planning accurate arm movements in a pointing task with the head at different orientations. When asked to point at an object, subjects oriented their arm to position the fingertip on the line running from the target to the viewing eye. But this eye-target line shifts when the eyes translate with each new head orientation, thereby requiring a new arm pointing direction. We confirmed that subjects do realign their fingertip with the eye-target line during closed-loop pointing across various horizontal head orientations when gaze is on target. More importantly, subjects also showed this head-position-dependent pattern of pointing responses for the same paradigm performed in complete darkness. However, when gaze was not on target, compensation for these translations in the rotational centers partially broke down. As a result, subjects tended to overshoot the target direction relative to current gaze; perhaps explaining previously reported errors in aiming the arm to retinally peripheral targets. These results suggest that knowledge of head position signals and the resulting relative displacements in the centers of rotation of the eye and shoulder are incorporated using open-loop mechanisms for eye-hand coordination, but these translations are best calibrated for foveated, gaze-on-target movements. PMID- 11929891 TI - Na(+)/Ca(2+) exchanger in GABAergic presynaptic boutons of rat central neurons. AB - Rat Meynert neurons were acutely isolated using a dissociation technique that maintains functional GABAergic presynaptic boutons. Miniature inhibitory postsynaptic currents (mIPSCs) were recorded under voltage-clamp conditions using whole cell patch-clamp recordings. Using the frequency of mIPSCs as a measure of presynaptic terminal excitability, the existence of a Na(+)/Ca(2+) exchanger (NCX) in these GABAergic nerve terminals was clearly demonstrated. Both the frequency and the amplitude of mIPSCs were unaffected by replacement of extracellular Na(2+). However, in this Na(+)-free external solution, ouabain could now induce a transient increase of mIPSCs frequency, which was not inhibited by adding Cd(2+) or cyclopiazonic acid but was inhibited by removing external Ca(2+). This indicates that this transient potentiation was dependent on external Ca(2+), but that this Ca(2+) influx was not via voltage-dependent Ca(2+) channels. KB-R7943, an inhibitor of NCX, at a concentration of 3 x 10(-6) M, reduced this transient increase of mIPSCs frequency without affecting mIPSCs amplitude and the response to exogenous GABA. These results demonstrate the existence of NCX in these GABAergic nerve terminals. In zero external Na(+), ouabain causes an accumulation of intraterminal Na(+) and a resultant influx of Ca(2+) through the reversed mode operation of NCX. However, under more physiological conditions, NCX may also operate in a forward mode and serve to maintain low intracellular [Ca(2+)] in nerve terminals. PMID- 11929892 TI - Discharge patterns of hypoglossal motoneurons during fictive breathing, coughing, and swallowing. AB - We performed a series of experiments to study the intracellular activity of 58 hypoglossal motoneurons (HMs) in decerebrate, paralyzed, and ventilated cats. Changes in membrane potentials (MP) and discharge activities were evaluated during fictive breathing (FB), swallowing (FS), and coughing (FC). FS and FC were elicited by electrical stimulation of the superior laryngeal nerves. FB, FS, and FC all exhibited characteristic discharge patterns of the phrenic, abdominal, pharyngeal branch of the vagus, and hypoglossal nerves. Thirty-nine HMs displayed respiratory modulation, and 19 were nonrespiratory modulated. Nine HMs did not exhibit MP changes during FB, FS, and FC. During FS, 49 HMs exhibited MP changes consisting of depolarization, hyperpolarization or hyperpolarization depolarization. HMs involved in FS were either respiratory modulated (n = 38) or not (n = 11). Only 20 HMs displayed MP changes and/or discharge activity during FC. All but two HMs fired during the expiratory phase of FC or at the end of this reflex. All HMs involved in FC (n = 20) were also modulated during both FB and FS. Our results suggest that the XII nucleus is functionally divided into common and distinct subsets of HMs based on their spontaneous activities and responses observed during FS and FC. The changes in MP and discharge frequencies observed during the three behaviors also suggest that HMs are driven by specific premotor neurons during FS, whereas a common premotor pathway is involved during FB and FC. PMID- 11929893 TI - Effect of the glial envelope on extracellular K(+) diffusion in olfactory glomeruli. AB - In many species, including vertebrates and invertebrates, first-order olfactory neuropils are organized into spherical glomeruli, partially enveloped by glial borders. The effect of this characteristic organization on olfactory information processing is poorly understood. The extracellular concentration of potassium ions ([K(+)]) must rise around olfactory receptor axons in specific glomeruli following odor-induced activation. To explore the time course and magnitude of K(+) accumulation and possible effects of such accumulation on neural activity within and among glomeruli, we developed a theoretical model to simulate the diffusion of K(+) in extracellular spaces of the glomeruli of the moth Manduca sexta. K(+) released by activated axons was assumed to diffuse through the extracellular spaces in glomeruli and the glial borders that surround them. The time-dependent diffusion equations were solved in spherical coordinates using a finite-difference method. The results indicate that the glial envelope forms a significant barrier to the spread of K(+) between neighboring glomeruli, thus reducing the likelihood of cross-talk between glomeruli, and may cause elevation of extracellular [K(+)] to levels that influence neural activity within the activated glomerulus for many seconds. Such effects could enhance olfactory discrimination and sensitivity, respectively. PMID- 11929894 TI - Representation of spectral and temporal envelope of twitter vocalizations in common marmoset primary auditory cortex. AB - Cortical sensitivity in representations of behaviorally relevant complex input signals was examined in recordings from primary auditory cortical neurons (AI) in adult, barbiturate-anesthetized common marmoset monkeys (Callithrix jacchus). We studied the robustness of distributed responses to natural and degraded forms of twitter calls, social contact vocalizations comprising several quasi-periodic phrases of frequency and AM. We recorded neuronal responses to a monkey's own twitter call (MOC), degraded forms of their twitter call, and sinusoidal amplitude modulated (SAM) tones with modulation rates similar to those of twitter calls. In spectral envelope degradation, calls with narrowband channels of varying bandwidths had the same temporal envelope as a natural call. However, the carrier phase was randomized within each narrowband channel. In temporal envelope degradation, the temporal envelope within narrowband channels was filtered while the carrier frequencies and phases remained unchanged. In a third form of degradation, noise was added to the natural calls. Spatiotemporal discharge patterns in AI both within and across frequency bands encoded spectrotemporal acoustic features in the call although the encoded response is an abstract version of the call. The average temporal response pattern in AI, however, was significantly correlated with the average temporal envelope for each phrase of a call. Response entrainment to MOC was significantly correlated with entrainment to SAM stimuli at comparable modulation frequencies. Sensitivity of the response patterns to MOC was substantially greater for temporal envelope than for spectral envelope degradations. The distributed responses in AI were robust to additive continuous noise at signal-to-noise ratios > or =10 dB. Neurophysiological data reflecting response sensitivity in AI to these forms of degradation closely parallel human psychophysical results on the intelligibility of degraded speech in quiet and noisy conditions. PMID- 11929895 TI - Differences in mechano-transducer channel kinetics underlie tonotopic distribution of fast adaptation in auditory hair cells. AB - The first step in audition is a deflection of the sensory hair bundle that opens mechanically gated channels, depolarizing the sensory hair cells. Two mechanism of adaptation of mechano-electric transducer (MET) channels have been identified in turtle auditory hair cells. The rate of fast adaptation varies tonotopically and is postulated to underlie a mechanical tuning mechanism in turtle auditory hair cells. Fast adaptation is driven by a calcium-dependent feedback process associated with MET channels. The purpose of this paper is to test the hypothesis that fast adaptation contributes to MET channel kinetics and that variations in channel kinetics underlie the tonotopic distribution of fast adaptation. To test for kinetic differences, the open channel blocker dihydrostreptomycin (DHS) was used. DHS blocked MET currents from low-frequency cells (IC(50) = 14 +/- 2 microM) more effectively than high-frequency cells (IC(50) = 75 +/- 5 microM), suggesting differences in MET channel properties. DHS block showed similar calcium sensitivities at both papilla locations. No difference in calcium permeation or block of the transducer channels was observed, indicating that the DHS effect was not due to differences in the channel pore. Slowing adaptation increased DHS efficacy, and speeding adaptation decreased DHS efficacy, suggesting that adaptation was influencing DHS block. DHS block of MET channels slowed adaptation, most likely by reducing the peak intraciliary calcium concentration achieved, supporting the hypothesis that the rate of adaptation varies with the calcium load per stereocilia. Another channel blocker, amiloride showed similar efficacy for high- and low-frequency cells with an IC(50) of 24.2 +/- 0.5 microM and a Hill coefficient of 2 but appeared to block high-frequency channels faster than low-frequency channels. To further explore MET channel differences between papilla locations, stationary noise analysis was performed. Spectral analysis of the noise gave half power frequencies of 1,185 +/- 148 Hz (n = 6) and 551 +/- 145 Hz (n = 5) for high- and low-frequency cells in 2.8 mM external calcium. The half power frequency showed similar calcium sensitivity to that of adaptation shifting to 768 +/- 205 Hz (n = 4) and 289 +/- 63 Hz (n = 4) for high- and low-frequency cells in 0.25 mM external calcium. Both the pharmacological data and the noise analysis data are consistent with the hypothesis that the tonotopic distribution of fast adaptation is in part due to differences in MET channel kinetics. An increase in the number of MET channels per stereocilia (termed summation) and or intrinsic differences in MET channel kinetics may be the underlying mechanism involved in establishing the gradient. PMID- 11929896 TI - Cortical representation of auditory space: information-bearing features of spike patterns. AB - Previous studies have demonstrated that the spike patterns of cortical neurons vary systematically as a function of sound-source location such that the response of a single neuron can signal the location of a sound source throughout 360 degrees of azimuth. The present study examined specific features of spike patterns that might transmit information related to sound-source location. Analysis was based on responses of well-isolated single units recorded from cortical area A2 in alpha-chloralose-anesthetized cats. Stimuli were 80-ms noise bursts presented from loudspeakers in the horizontal plane; source azimuths ranged through 360 degrees in 20 degrees steps. Spike patterns were averaged across samples of eight trials. A competitive artificial neural network (ANN) identified sound-source locations by recognizing spike patterns; the ANN was trained using the learning vector quantization learning rule. The information about stimulus location that was transmitted by spike patterns was computed from joint stimulus-response probability matrices. Spike patterns were manipulated in various ways to isolate particular features. Full-spike patterns, which contained all spike-count information and spike timing with 100-micros precision, transmitted the most stimulus-related information. Transmitted information was sensitive to disruption of spike timing on a scale of more than approximately 4 ms and was reduced by an average of approximately 35% when spike-timing information was obliterated entirely. In a condition in which all but the first spike in each pattern were eliminated, transmitted information decreased by an average of only approximately 11%. In many cases, that condition showed essentially no loss of transmitted information. Three unidimensional features were extracted from spike patterns. Of those features, spike latency transmitted approximately 60% more information than that transmitted either by spike count or by a measure of latency dispersion. Information transmission by spike patterns recorded on single trials was substantially reduced compared with the information transmitted by averages of eight trials. In a comparison of averaged and nonaveraged responses, however, the information transmitted by latencies was reduced by only approximately 29%, whereas information transmitted by spike counts was reduced by 79%. Spike counts clearly are sensitive to sound-source location and could transmit information about sound-source locations. Nevertheless, the present results demonstrate that the timing of the first poststimulus spike carries a substantial amount, probably the majority, of the location-related information present in spike patterns. The results indicate that any complete model of the cortical representation of auditory space must incorporate the temporal characteristics of neuronal response patterns. PMID- 11929897 TI - Changes in crossed spinal reflexes after peripheral nerve injury and repair. AB - We investigated the changes induced in crossed extensor reflex responses after peripheral nerve injury and repair in the rat. Adults rats were submitted to non repaired sciatic nerve crush (CRH, n = 9), section repaired by either aligned epineurial suture (CS, n = 11) or silicone tube (SIL4, n = 13), and 8 mm resection repaired by tubulization (SIL8, n = 12). To assess reinnervation, the sciatic nerve was stimulated proximal to the injury site, and the evoked compound muscle action potential (M and H waves) from tibialis anterior and plantar muscles and nerve action potential (CNAP) from the tibial nerve and the 4th digital nerve were recorded at monthly intervals for 3 mo postoperation. Nociceptive reinnervation to the hindpaw was also assessed by plantar algesimetry. Crossed extensor reflexes were evoked by stimulation of the tibial nerve at the ankle and recorded from the contralateral tibialis anterior muscle. Reinnervation of the hindpaw increased progressively with time during the 3 mo after lesion. The degree of muscle and sensory target reinnervation was dependent on the severity of the injury and the nerve gap created. The crossed extensor reflex consisted of three bursts of activity (C1, C2, and C3) of gradually longer latency, lower amplitude, and higher threshold in control rats. During follow-up after sciatic nerve injury, all animals in the operated groups showed recovery of components C1 and C2 and of the reflex H wave, whereas component C3 was detected in a significantly lower proportion of animals in groups with tube repair. The maximal amplitude of components C1 and C2 recovered to values higher than preoperative values, reaching final levels between 150 and 245% at the end of the follow-up in groups CRH, CS, and SIL4. When reflex amplitude was normalized by the CNAP amplitude of the regenerated tibial nerve, components C1 (300-400%) and C2 (150-350%) showed highly increased responses, while C3 was similar to baseline levels. In conclusion, reflexes mediated by myelinated sensory afferents showed, after nerve injuries, a higher degree of facilitation than those mediated by unmyelinated fibers. These changes tended to decline toward baseline values with progressive reinnervation but still remained significant 3 mo after injury. PMID- 11929898 TI - Quantitative analysis of catch-up saccades during sustained pursuit. AB - During visual tracking of a moving stimulus, primates orient their visual axis by combining two very different types of eye movements, smooth pursuit and saccades. The purpose of this paper was to investigate quantitatively the catch-up saccades occurring during sustained pursuit. We used a ramp-step-ramp paradigm to evoke catch-up saccades during sustained pursuit. In general, catch-up saccades followed the unexpected steps in position and velocity of the target. We observed catch-up saccades in the same direction as the smooth eye movement (forward saccades) as well as in the opposite direction (reverse saccades). We made a comparison of the main sequences of forward saccades, reverse saccades, and control saccades made to stationary targets. They were all three significantly different from each other and were fully compatible with the hypothesis that the smooth pursuit component is added to the saccadic component during catch-up saccades. A multiple linear regression analysis was performed on the saccadic component to find the parameters determining the amplitude of catch-up saccades. We found that both position error and retinal slip are taken into account in catch-up saccade programming to predict the future trajectory of the moving target. We also demonstrated that the saccadic system needs a minimum period of approximately 90 ms for taking into account changes in target trajectory. Finally, we reported a saturation (above 15 degrees /s) in the contribution of retinal slip to the amplitude of catch-up saccades. PMID- 11929899 TI - Adenosine acting on A1 receptors protects NO-triggered rebound potentiation and LTP in rat hippocampal slices. AB - Exposure of hippocampal slices to nitric oxide (NO) results in a depression of CA1 synaptic transmission. Under 0.2-Hz stimulation, washout of NO leads to a persistent potentiation that depends on N-methyl-D-aspartate (NMDA) receptors and endogenous NO formation and that occludes tetanus-induced long-term potentiation (LTP). The experiments were initially aimed at determining the relationship between the NO-induced synaptic depression and rebound potentiation. The adenosine A1 antagonist, 8-cyclopentyl-1,3-dipropylxanthine (DPCPX) partially inhibited the depression produced by the NO donor diethylamine NONOate (300 microM). It also led to a complete block of both the rebound potentiation and the subsequent tetanus-induced LTP. LTP was preserved in the presence of DPCPX if the stimulation frequency was reduced to 0.033 Hz or if the NO application was omitted. The NO-triggered rebound potentiation was restored if the experiment (DPCPX followed by exogenous NO) was conducted in the presence of an NMDA antagonist. The restored potentiation was completely blocked by the NO synthase inhibitor, L-nitroarginine. It is concluded that the NO-induced depression is partially mediated by increased release of endogenous adenosine acting on A1 receptors. Moreover, tonic A1 receptor activation by adenosine protects LTP and the rebound potentiation from being disabled by untimely NMDA receptor activity. Hence, the NO-induced depression and rebound potentiation are linked in the sense that the depression helps to preserve the capacity of the synapses to undergo potentiation. Finally, the results give the first example of exogenous NO eliciting an enduring potentiation of hippocampal synaptic transmission that is dependent on endogenous NO formation, but not on NMDA receptors. PMID- 11929900 TI - Evidence that REM sleep is controlled by the activation of brain stem pedunculopontine tegmental kainate receptor. AB - Glutamate, the neurotransmitter, enhances rapid-eye-movement (REM) sleep when microinjected into the brain stem pedunculopontine tegmentum (PPT) of the cat and rat. Glutamate and its various receptors are normally present in the PPT cholinergic cell compartment. The aim of this study was to identify which specific receptor(s) in the cholinergic cell compartment of the PPT are involved in glutamate-induced-REM sleep. To identify these glutamate-induced REM-sleep generating receptor(s) in the PPT cholinergic cell compartment, specific receptors were pharmacologically blocked differentially by localized pretreatment of specific glutamate receptor antagonists; glutamate was then microinjected into the PPT cholinergic cell compartment while quantifying the effects on REM sleep in freely moving chronically instrumented rats. The results demonstrate that when kainate receptors were blocked by pretreatment with a kainate-specific receptor antagonist, microinjection of glutamate was unable to induce REM sleep. Pharmacological blockade of specific N-methyl-D-aspartate and alpha-amino-3 hydroxy-5-methyl-4-isoxazolepropionic acid receptors was unable to block glutamate-microinjection-induced-REM sleep. These findings suggest, for the first time, that the activation of kainate receptors within the cholinergic cell compartment of the PPT is an essential portion of the mechanism for the generation of glutamate-induced REM sleep in the rat. PMID- 11929901 TI - Threshold conditions for synaptically evoking Ca(2+) waves in hippocampal pyramidal neurons. AB - Regenerative Ca(2+) release from inositol 1,4,5-trisphosphate (IP(3))-sensitive intracellular stores in the form of Ca(2+) waves leads to large-amplitude [Ca(2+)](i) increases in the apical dendrites of hippocampal CA1 pyramidal neurons. Release is generated following synaptic activation of group I metabotropic glutamate (mGlu) receptors. We systematically examined the conditions for evoking these waves in transverse slices from 2- to 3-wk-old rats. Using a sharpened asymmetrical bipolar tungsten stimulating electrode placed in the stratum radiatum, we varied the lateral position of the electrode, the number of stimulating pulses, the train frequency, and stimulus current. Several trends were clear. Increasing the frequency of stimulation from 20 to 100 Hz, keeping the total number of pulses constant, lowered the required stimulus current. Stimulation at frequencies below 20 Hz made it difficult to evoke release. Increasing the number of stimulation pulses, keeping the frequency constant, lowered the threshold current. A minimum of five pulses at 100 Hz was required to evoke release reliably, but several examples of success with three pulses were recorded. Theta-burst stimulation was as effective as tetanic stimulation. Placing the point of the stimulation electrode closer to the pyramidal neuron made it easier to evoke release, although stimulation at a lateral distance of 500 microm with unsharpened electrodes was sometimes successful. The simplest explanation for these results is that a bolus of IP(3) must be produced quickly in a restricted region of the dendrites to generate Ca(2+) waves. The conditions necessary for evoking regenerative Ca(2+) release have many parallels (and some differences) with the conditions required to evoke long-term potentiation in these cells following tetanic stimulation. PMID- 11929902 TI - Superior colliculus activity related to concurrent processing of saccade goals in a visual search task. AB - Saccades are typically separated by inter-saccadic fixation intervals (ISFIs) of > or =125 ms. During this time, the saccadic system selects a goal and completes the preparatory processes required prior to executing the subsequent movement. However, in tasks in which competing stimuli are presented, two sequentially executed movements to different goals can be separated by much shorter ISFIs. This suggests that the saccadic system is capable of completing many of the preparatory requirements for a second saccade concurrently with the execution of an initial movement. We recorded single neurons in the superior colliculus (SC) during rapid saccade sequences made by rhesus monkeys performing a search task. We found that during the execution of an initial saccade, activity related to the goal of a quickly following second saccade can be simultaneously maintained in the SC motor map. This activity appears to signal the selection or increased salience of the second saccade goal even before the initial saccade has ended. For movements separated by normal ISFIs (> or =125 ms), we did not observe activity related to concurrent processing, presumably because for these longer ISFI responses, the goal of the second saccade is not selected until after the end of the first saccade. These results indicate that, at the time of an initial saccade, the SC does not necessarily act as a strict winner-take-all network. Rather it appears that the salience of a second visual goal can be simultaneously maintained in the SC. This provides evidence that selection or preparatory activity related to the goal of a second saccade can overlap temporally with activity related to an initial saccade and indicates that such concurrent processing is present even in a structure which is fairly close to the motor output. PMID- 11929903 TI - Ca(2+)-dependent Ca(2+) clearance via mitochondrial uptake and plasmalemmal extrusion in frog motor nerve terminals. AB - Ca(2+) clearance in frog motor nerve terminals was studied by fluorometry of Ca(2+) indicators. Rises in intracellular Ca(2+) ([Ca(2+)](i)) in nerve terminals induced by tetanic nerve stimulation (100 Hz, 100 or 200 stimuli: Ca(2+) transient) reached a peak or plateau within 6-20 stimuli and decayed at least in three phases with the time constants of 82-87 ms (81-85%), a few seconds (11 12%), and several tens of seconds (less than a few percentage). Blocking both Na/Ca exchangers and Ca(2+) pumps at the cell membrane by external Li(+) and high external pH (9.0), respectively, increased the time constants of the initial and second decay components with no change in their magnitudes. By contrast, similar effects by Li(+) alone, but not by high alkaline alone, were seen only on 200 stimuli-induced Ca(2+) transients. Blocking Ca(2+) pumps at Ca(2+) stores by thapsigargin did not affect 100 stimuli-induced Ca(2+) transients but increased the initial decay time constant of 200 stimuli-induced Ca(2+) transients with no change in other parameters. Inhibiting mitochondrial Ca(2+) uptake by carbonyl cyanide m-chlorophenylhydrazone markedly increased the initial and second decay time constants of 100 stimuli-induced Ca(2+) transients and the amplitudes of the second and the slowest components. Plotting the slopes of the decay of 100 stimuli-induced Ca(2+) transients against [Ca(2+)](i) yielded the supralinear [Ca(2+)](i) dependence of Ca(2+) efflux out of the cytosol. Blocking Ca(2+) extrusion or mitochondrial Ca(2+) uptake significantly reduced this [Ca(2+)](i) dependent Ca(2+) efflux. Thus Ca(2+)-dependent mitochondrial Ca(2+) uptake and plasmalemmal Ca(2+) extrusion clear out a small Ca(2+) load in frog motor nerve terminals, while thapsigargin-sensitive Ca(2+) pump boosts the clearance of a heavy Ca(2+) load. Furthermore, the activity of plasmalemmal Ca(2+) pump and Na/Ca exchanger is complementary to each other with the slight predominance of the latter. PMID- 11929904 TI - Evidence of a functionally segregated pathway from dorsal cochlear nucleus to inferior colliculus. AB - Type O units in the central nucleus of the inferior colliculus (ICC) of decerebrate cats are excited by best frequency (BF) tones near threshold, but are inhibited by high-level tones at all frequencies. Dorsal cochlear nucleus (DCN) principal cells display similar response map features and project directly to the ICC, and are thus supposed to be the dominant source of excitatory input for type O units. To test this hypothesis, the responses of type O units were compared before and after two pharmacological manipulations. When DCN to ICC axons were blocked by pressure injections of lidocaine, most type O units (approximately 80%) were silenced or showed substantially reduced activity, but some units showed increased activity. All of the former units had low maximal rates to BF tones, whereas the latter units had high rates. When local circuit inhibitory mechanisms in the ICC were blocked by iontophoretic application of bicuculline or strychnine, type O unit responses also fell into two classes: low-rate units that showed increased spontaneous and driven activities and high-rate units that showed, in addition, altered response map features. Taken together, these results demonstrate that low-rate type O units are part of a functionally segregated pathway initiated by the DCN, whereas high-rate type O units are created at the level of the ICC. PMID- 11929906 TI - Intrinsic activation of human motoneurons: possible contribution to motor unit excitation. AB - The main purpose of this study was to estimate the contribution of intrinsic activation of human motoneurons (e.g., by plateau potentials) during voluntary and reflexive muscle contractions. Pairs of motor units were recorded from either the tibialis anterior or soleus muscle during three different conditions: 1) during a brief muscle vibration followed by a slow relaxation of a steady isometric contraction; 2) during a triangular isometric torque contraction; and 3) during passive sinusoidal muscle stretch superimposed on a steady isometric contraction. In each case, the firing rate of a tonically firing control motor unit was used as a measure of the effective synaptic excitation (i.e., synaptic drive) to a slightly higher-threshold test motor unit that was recruited and de recruited during a contraction trial. The firing rate of the control unit was compared at recruitment and de-recruitment of the test unit. This was done to determine whether the estimated synaptic drive needed to recruit a motor unit was less than the amount needed to sustain firing as a result of an added depolarization produced from intrinsic sources. After test unit recruitment, the firing rate of the control unit could be decreased significantly (on average by 3.6 Hz from an initial recruitment rate of 9.8 Hz) before the test unit was de recruited during a descending synaptic drive. Similar decreases in control unit rate occurred in all three experimental conditions. This represents a possible 40% reduction in the estimated synaptic drive needed to maintain firing of a motor unit compared with the estimated amount needed to recruit the unit initially. The firing rates of both the control and test units were modulated together in a highly parallel fashion, suggesting that the unit pairs were driven by common synaptic inputs. This tight correlation further validated the use of the control unit firing rate as a monitor of synaptic drive to the test motor unit. The estimates of intrinsically mediated depolarization of human motoneurons ( approximately 40% during moderate contractions) are consistent with values obtained for plateau potentials obtained from intracellular recordings of motoneurons in reduced animal preparations, although various alternative mechanisms are discussed. This suggests that similar intrinsic conductances provide a substantial activation of human motoneurons during moderate physiological activity. PMID- 11929905 TI - Purkinje cells of the cerebellar dorsal vermis: simple-spike activity during pursuit and passive whole-body rotation. AB - To track a slowly moving object during whole body rotation, smooth-pursuit and vestibularly induced eye movements must interact to maintain the accuracy of eye movements in space (i.e., gaze), and gaze movement signals must eventually be converted into eye movement signals in the orbit. To understand the role played by the cerebellar vermis in pursuit-vestibular interactions, in particular whether the output of the vermis codes gaze-velocity or eye-velocity, we examined simple-spike activity of 58 Purkinje (P-) cells in lobules VI-VII of head stabilized Japanese monkeys that were trained to elicit smooth-pursuit eye movements and cancel their vestibuloocular reflex (VOR) during passive whole body rotation around horizontal, vertical, or oblique axes. All pursuit-sensitive vermal P-cells also responded during VOR cancellation, and the majority of them had peak modulation near peak stimulus velocity. The directions of maximum modulation during these two tasks were distributed in all directions with a downward preponderance. Using standard criteria, 40% of pursuit-sensitive vermal P-cells were classified as gaze-velocity. Other P-cells were classified either as eye/head-velocity group I (36%) that had similar preferred directions during pursuit and VOR cancellation but that had larger responses during VOR x1 when gaze remained stationary, or as eye/head-velocity group II (24%) that had oppositely directed or orthogonal eye and head movement sensitivity during pursuit and VOR cancellation. Eye/head-velocity group I P-cells contained cells whose activity was correlated with eye velocity. Modulation of many P-cells of the three groups during VOR x1 could be accounted for by the linear addition of their modulations during pursuit and VOR cancellation. When monkeys fixated a stationary target, over half of the P-cells tested, including gaze-velocity P cells, discharged in proportion to the velocity of retinal motion of a second spot. These observations are in a striking contrast to our previous results for floccular vertical P-cells. Because we used identical tasks, these differences suggest that the two cerebellar regions are involved in very different kinds of processing of pursuit-vestibular interactions. PMID- 11929907 TI - Intrinsic activation of human motoneurons: reduction of motor unit recruitment thresholds by repeated contractions. AB - The main purpose of this study was to examine whether facilitation of human motor unit recruitment by repeated voluntary contractions is mediated, in part, by time and activity-dependent increases in the intrinsic excitability of the parent motoneuron. To do this, pairs of tibialis anterior or soleus motor units were recorded during slowly increasing and then decreasing voluntary contractions. The firing rate of the lower-threshold motor unit of the pair (control unit) was used as a measure of effective synaptic excitation (i.e., drive) to the motoneurons. This rate was used to estimate the recruitment threshold of the higher-threshold unit of the pair (test unit). The test unit was repeatedly recruited and de recruited in a series of contractions, and the interval between the de recruitment and re-recruitment of the test unit (interactivation interval) was systematically varied between 0.6 and 60 s. An increase in intrinsic excitability of a unit was considered to have occurred if the level of estimated synaptic input (as measured by the firing rate of the control motor unit) needed to recruit a unit was reduced. At short interactivation intervals (1-2 s), the control unit firing frequency was significantly lower when the test unit was recruited on the second contraction, compared with the first (by 3.9 Hz or a 64% reduction). This suggested that the intrinsic excitability of the test motor unit had increased during the second contraction because it could be recruited at a much lower level of estimated synaptic drive. Longer interaction intervals (2-6 s) produced less recruitment facilitation. At even longer interactivation intervals (>6 s) there was no significant facilitation (time constant of effect was 4.8 s). In some motor units, the effect of this short-term facilitation appeared to be so pronounced that it resulted in reversing the order of de recruitment with the other initially lower-threshold motor units. Such reversals were occasionally observed for orderly re-recruitment. The time course and behavior of the observed short-term facilitation of motor unit discharge was qualitatively similar to the warm-up phenomenon of plateau potentials seen in motoneurons of reduced preparations (e.g., 4-6 s). The possibility of warm-up contributing to the time and activity-dependent facilitation of human motor unit recruitment is discussed. PMID- 11929908 TI - Physiological correlates of perceptual learning in monkey V1 and V2. AB - Performance in visual discrimination tasks improves with practice. Although the psychophysical parameters of these improvements have suggested the involvement of early areas in visual cortex, there has been little direct study of the physiological correlates of such perceptual learning at the level of individual neurons. To examine how neuronal response properties in the early visual system may change with practice, we trained monkeys for more than 6 mo in an orientation discrimination task in which behaviorally relevant stimuli were restricted to a particular retinal location and oriented around a specific orientation. During training the monkeys' discrimination thresholds gradually improved to much better than those of naive monkeys or humans. Although this improvement was specific to the trained orientation, it showed little retinotopic specificity. The receptive field properties of single neurons from regions representing the trained location and a location in the opposite visual hemifield were measured in V1 and V2. In most respects the receptive field properties in the representations of the trained and untrained regions were indistinguishable. However, in the regions of V1 and V2 representing the trained location, there were slightly fewer neurons whose optimal orientation was near the trained orientation. This resulted in a small but significant decrease in the V1 population response to the trained orientation at the trained location. Consequently, the observed neuronal populations did not exhibit any orientation-specific biases sufficient to explain the orientation specificity of the behavioral improvement. Pooling models suggest that the behavioral improvement was accomplished with a task-dependent and orientation-selective pooling of unaltered signals from early visual neurons. These data suggest that, even for training with stimuli suited to the selectivities found in early areas of visual cortex, behavioral improvements can occur in the absence of pronounced changes in the physiology of those areas. PMID- 11929909 TI - Responses of spinothalamic lamina I neurons to maintained noxious mechanical stimulation in the cat. AB - Noxious mechanical stimuli that are maintained for minutes produce a continuous sensation of pain in humans that augments during the stimulus. It has recently been shown with systematic force-controlled stimuli that, while all mechanically responsive nociceptors adapt to these stimuli, the basis for such pain can be ascribed to A-fiber rather than C-fiber nociceptors, based on distinctions in their respective response profiles and stimulus-response functions. The present experiments investigated whether similar distinctions could be made in subsets of nociceptive lamina I spinothalamic tract (STT) neurons using similar maintained stimuli. Twenty-eight lamina I STT neurons in the lumbosacral dorsal horn of barbiturate-anesthetized cats were tested with noxious mechanical stimuli applied with a probe of 0.1 mm(2) contact area at forces of 25, 50, and 100 g for 2 min. The neurons were classified as nociceptive-specific (NS, n = 14) or polymodal nociceptive (HPC, n = 14) based on their responses to quantitative thermal stimuli. The NS neurons had greater responses and showed less adaptation than the HPC neurons in response to these stimuli, and they encoded stimulus intensity better. Comparison of the normalized response profiles of all 28 nociceptive lamina I STT neurons, independent of cell classification, revealed 2 subgroups that differed significantly: "Maintained" cells with responses that remained above 50% of the initial peak rate during stimulation and "Adapting" cells with responses that quickly declined to <50%. The Maintained neurons encoded the intensity of the mechanical stimuli better than the Adapting neurons, based on ratiometric functions. A k-means cluster analysis of all 28 cells distinguished the identical two subgroups. These categories corresponded closely to the NS and HPC categories: Maintained cells were mostly NS neurons (10 NS, 3 HPC), and Adapting cells were mostly HPC neurons (4 NS, 11 HPC). Thus the present data are consistent with the distinctions between A-fiber and C-fiber nociceptors observed previously, because A-fiber nociceptors are the predominant input to NS lamina I STT neurons and C-fiber nociceptors are the predominant input to HPC neurons. These findings support the view that NS, but perhaps not HPC, lamina I STT neurons have a role in the pain caused by maintained mechanical stimuli and contribute to the sensations of "first" pain and "sharpness." Nonetheless, none of the units studied showed increasing responses during the stimuli, suggesting a role for other ascending neurons or forebrain integration in the augmenting pain produced by maintained mechanical stimulation. PMID- 11929910 TI - Responses of spinothalamic lamina I neurons to repeated brief contact heat stimulation in the cat. AB - It was recently shown that repeated heat stimulation, using brief contacts (<1 s) with a preheated thermode at sufficiently short interstimulus intervals (ISIs <5 s) and high temperatures (> or =51 degrees C), will elicit in humans a sensation of rapidly augmenting "second" (burning) pain with only a weak "first" (sharp) pain sensation. Most strikingly, at short intertrial intervals (ITIs >5 s) such summation will reset, or begin again at baseline. In the present experiments, the responses of nociceptive lamina I spinothalamic (STT) neurons in the lumbosacral dorsal horn of barbiturate-anesthetized cats were examined using this repeated brief contact heat paradigm. The neurons were classified as nociceptive-specific (NS, n = 8) or polymodal nociceptive (HPC, n = 8) based on their responses to quantitative thermal stimuli; all had receptive fields on the glabrous ventral hindpaw. A pneumatic piston was used to apply a thermode preheated to 34, 46, 49, 53, or 58 degrees C with a contact dwell time of approximately 0.7 s to the ventral hindpaw repeatedly (15 times) at ISIs of 2, 3, and 5 s, with 3-5 min between trials. The mean responses of the 16 nociceptive lamina I STT cells showed rapid temporal summation that was directly dependent on temperature and inversely dependent on ISI, with the greatest increases occurring between the 3rd and 10th contacts. The temporal profiles of this family of curves correspond with the psychophysical data on human sensation. Further analysis showed that this summation was due to the HPC cells, which all showed strong summation; in contrast, the NS cells showed little, if any. The HPC responses to the repeated heat stimuli lagged each contact by approximately 1 s, consistent with the strong, monosynaptic C-fiber input that is characteristic of HPC cells and also with the dependence of second pain on C-fiber nociceptors. HPC cells also displayed the reset phenomenon at short ITIs, again in correspondence with the psychophysical data. The summation and the reset displayed by HPC cells were not related to skin temperature. Thus the results presented in this study, together with those in the preceding article, demonstrate a double dissociation indicating that NS and HPC lamina I STT cells can subserve the qualitatively distinct sensations of first (sharp) and second (burning) pain, respectively. These findings support the concept that the lamina I STT projection comprises several discrete sensory channels that are integrated in the forebrain to generate distinct sensations. PMID- 11929911 TI - Effect of short-term saccadic adaptation on saccades evoked by electrical stimulation in the primate superior colliculus. AB - The brain maintains the accuracy of visually guided movements by using visual feedback to correct for changes in the nervous system and musculature that would otherwise result in dysmetria. In monkeys, evidence suggests that an adaptive mechanism can compensate for weakness in an extraocular muscle by changing the gain of the neural signal to the weakened muscle. The visual effects of such neuromuscular changes have been simulated using a short-term saccade adaptation paradigm, in which the target spot jumps to a new location during the initial saccade. Under these circumstances, over several hundred trials, monkeys gradually change the amplitude of their saccades so that the eye lands closer to the final location of the target spot. There is considerable evidence from lesion and single-unit recording studies that the locus of such saccade adaptation is downstream of the superior colliculus in the cerebellum. Paradoxically, previous research has indicated that saccades evoked by electrical stimulation in the superior colliculus are not modified by short-term saccade adaptation, suggesting that adaptation occurs in the oculomotor system upstream of the superior colliculus or else in a pathway that bypasses the superior colliculus. We tested whether this result was due to using suprathreshold stimulation currents. Stimulating at 44 low-threshold sites in the superior colliculi of three monkeys revealed that using low current levels evoked saccades that were modified by adaptation. Adaptation for visually guided and electrically evoked saccades had similar time courses and tended to be accomplished by a reduction in saccade velocity rather than a decrease in duration. Moreover, the more similar the velocity of electrically evoked and visually guided saccades prior to the start of saccadic adaptation the greater the effect of adaptation on electrically evoked saccades. These results suggest that the superior colliculus is indeed upstream of the locus of adaptation, corroborating previous lesion and single cell recording studies, but that the mechanism mediating saccade adaptation is sensitive to the parameters of electrical stimulation. PMID- 11929912 TI - Two different mechanisms underlie reversible, intrinsic optical signals in rat hippocampal slices. AB - Intrinsic optical signals (IOSs) induced by synaptic stimulation and moderate hypotonic swelling in brain tissue slices consist of reduced light scattering and are usually attributed to cell swelling. During spreading depression (SD), however, light-scattering increases even though SD has been shown to cause strong cell swelling. To understand this phenomenon, we recorded extracellular voltage, light transmission (LT), which is inversely related to light scattering, and interstitial volume (ISV) simultaneously from the same site (stratum radiatum of CA1) in both interface and submerged hippocampal slices. As expected, moderate lowering of bath osmolarity caused concentration-dependent shrinkage of ISV and increase in LT, while increased osmolarity induced opposite changes in both variables. During severe hypotonia, however, after an initial increase of LT, the direction of the IOS reversed to a progressive decrease in spite of continuing ISV shrinkage. SD caused by hypotonia, by microinjection of high-K(+) solution, or by hypoxia, was associated with a pronounced LT decrease, during which ISV shrinkage indicated maximal cell swelling. If most of the extracellular Cl(-) was substituted by the impermeant anion methylsulfate and also in strongly hypertonic medium, the SD-related decrease in LT was suppressed and replaced by a monotonic increase. Nevertheless, the degree of ISV shrinkage was similar in low and in normal Cl(-) conditions. The optical signals and ISV changes were qualitatively identical in interface and submerged slices. We conclude that there are at least two mechanisms that underlie reversible optical responses in hippocampal slices. The first mechanism underlies light-scattering decrease (hence enhancing LT) when ISV shrinks (cell swelling) under synaptic stimulation and mild hypotonia. Similarly, as result of this mechanism, expansion of ISV (cell shrinkage) during mild hypertonia leads to an increased light scattering (and decreased LT). Thus optical signals associated with this first mechanism show expected cell-volume changes and are linked to either cell swelling or shrinkage. A different mechanism causes the light-scattering increase (leading to a LT decrease) during severe hypotonia and various forms of SD but with a severely decreased ISV. This second mechanism may be due to organelle swelling or dendritic beading but not to cell-volume increase. These two mechanisms can summate, indicating that they are independent in origin. Suppression of the SD-related light-scattering increase by lowering [Cl(-)](o) or severe hypertonia unmasks the underlying swelling-related scattering decrease. The simultaneous IOS and ISV measurements clearly distinguish these two mechanisms of optical signal generation. PMID- 11929913 TI - Inhibition of glutamatergic synaptic input to spinal lamina II(o) neurons by presynaptic alpha(2)-adrenergic receptors. AB - Activation of spinal alpha(2)-adrenergic receptors by the descending noradrenergic system and alpha(2)-adrenergic agonists produces analgesia. However, the sites and mechanisms of the analgesic action of spinally administered alpha(2)-adrenergic receptor agonists such as clonidine are not fully known. The dorsal horn neurons in the outer zone of lamina II (lamina II(o)) are important for processing nociceptive information from C-fiber primary afferents. In the present study, we tested a hypothesis that activation of presynaptic alpha(2)-adrenergic receptors by clonidine inhibits the excitatory synaptic input to lamina II(o) neurons. Whole cell voltage-clamp recordings were performed on visualized lamina II(o) neurons in the spinal cord slice of rats. The miniature excitatory postsynaptic currents (mEPSCs) were recorded in the presence of tetrodotoxin, bicuculline, and strychnine. The evoked EPSCs were obtained by electrical stimulation of the dorsal root entry zone or the attached dorsal root. Both mEPSCs and evoked EPSCs were abolished by application of 6 cyano-7-nitroquinoxaline-2,3-dione. Clonidine (10 microM) significantly decreased the frequency of mEPSCs from 5.8 +/- 0.9 to 2.7 +/- 0.6 Hz (means +/- SE) without altering the amplitude and the decay time constant of mEPSCs in 25 of 27 lamina II(o) neurons. Yohimbine (2 microM, an alpha(2)-adrenergic receptor antagonist), but not prazosin (2 microM, an alpha(1)-adrenergic receptor antagonist), blocked the inhibitory effect of clonidine on the mEPSCs. Clonidine (1-20 microM, n = 8) also significantly attenuated the peak amplitude of evoked EPSCs in a concentration-dependent manner. The effect of clonidine on evoked EPSCs was abolished in the presence of yohimbine (n = 5). These data suggest that clonidine inhibits the excitatory synaptic input to lamina II(o) neurons through activation of alpha(2)-adrenergic receptors located on the glutamatergic afferent terminals. Presynaptic inhibition of glutamate release from primary afferents onto lamina II(o) neurons likely plays an important role in the analgesic action produced by activation of the descending noradrenergic system and alpha(2)-adrenergic agonists. PMID- 11929914 TI - NMDA receptor-mediated currents in rat cerebellar granule and unipolar brush cells. AB - The properties of N-methyl-D-aspartate (NMDA) receptor-mediated currents at the giant cerebellar mossy-fiber unipolar brush cell (UBC) synapse were compared with those of adjacent granule cells using patch-clamp recording methods in thin slices of rat cerebellar nodulus. In UBCs, NMDA receptor-mediated excitatory postsynaptic currents (EPSCs) decayed as a single exponential whose time constant was independent of membrane potential. The EPSC was reduced in all cells by the NR1/NR2B-selective antagonist ifenprodil, and the Zn(2+) chelator N,N,N',N' tetrakis(2-pyridylmethyl)ethylenediamine (TPEN) produced a transient potentiation in 50% of cells. In contrast, the NMDA EPSC in granule cells decayed as a double exponential that dramatically switched to a slower rate at positive membrane potentials. The synaptic response in some granule cells also displayed a late second peak at positive potentials, and in others, activation of mossy fibers produced repetitive trains of EPSCs indicating they may be postsynaptic to the UBC network. Single-channel recordings of outside-out somatic patches from UBCs in magnesium-free solution revealed only high-conductance (50 pS) channels whose open time was increased with depolarization, but the opening frequency was decreased to yield a low (p(o) = 0.0298), voltage-independent opening probability. Lowering extracellular calcium (2.5-0.25 mM) had no effects on channel gating, although an increase of single-channel conductance was observed at lower calcium concentrations. Taken together, the data support the notion that the NMDA receptor in UBCs may comprise both NR1/NR2A and NR1/NR2B receptors. Furthermore, the properties of the EPSC in these two classes of feedforward glutamatergic interneurons display fundamental differences that may relate to their roles in synaptic integration. PMID- 11929915 TI - Disparity-selective neurons in area V4 of macaque monkeys. AB - Area V4 is an intermediate stage of the ventral visual pathway providing major input to the final stages in the inferior temporal cortex (IT). This pathway is involved in the processing of shape, color, and texture. IT neurons are also sensitive to horizontal binocular disparity, suggesting that binocular disparity is processed along the ventral visual pathway. In the present study, we examined the processing of binocular disparity information by V4 neurons. We recorded responses of V4 neurons to binocularly disparate stimuli. A population of V4 neurons modified their responses according to changes of stimulus disparity; neither monocular responses nor eye movements could account for this modulation. Disparity-tuning curves were similar for different locations within a neuron's receptive field. Neighboring neurons recorded using a single electrode displayed similar disparity-tuning properties. These findings indicate that a population of V4 neurons is selective for binocular disparity, invariant for the position of the stimulus within the receptive field. The finding that V4 neurons with similar disparity selectivity are clustered suggests the existence of functional modules for disparity processing in V4. PMID- 11929916 TI - Glutamate uptake controls expression of a slow postsynaptic current mediated by mGluRs in cerebellar Purkinje cells. AB - At the cerebellar parallel fiber-Purkinje cell synapse, isolated presynaptic activity induces fast excitatory postsynaptic currents via ionotropic glutamate receptors while repetitive, high-frequency, presynaptic activity can also induce a slow excitatory postsynaptic current that is mediated by metabotropic glutamate receptors (mGluR1-EPSC). Here we investigated the involvement of glutamate uptake in the expression of the mGluR1-EPSC. Inhibitors of glutamate uptake led to a large increase of the mGluR1-EPSC. D-aspartate (0.4 mM) and L(-)-threo-3 hydroxyaspartate (0.4 mM) increased the mGluR1-EPSC approximately 4.5 and approximately 9-fold, respectively, while dihydrokainic acid (1 mM), had no significant effect on the mGluR1-EPSC. D-aspartate (0.4 mM) shifted the concentration-response curve of the depression of the mGluR1-EPSC by the low affinity mGluR1 antagonist (S)-a-Methyl-4-carboxyphenylglycine [(S)-MCPG] to higher concentrations and decreased the stimulus intensity and the number of necessary stimuli to evoke an mGluR1-EPSC. Depression of the mGluR1-EPSC by rapid pressure application of (S)-MCPG at varying time intervals after tetanic stimulation of the parallel fibers indicated that the glutamate concentration in the peri- and extrasynaptic space decayed with time constants of 36 and 316 ms under control conditions and with inhibition of glutamate uptake, respectively. These results show that expression of the slow mGluR-mediated excitatory postsynaptic current is controlled by glutamate transporter activity. Thus in contrast to fast glutamatergic synaptic transmission, metabotropic glutamate receptor-mediated transmission is critically dependent on the activity and capacity of glutamate uptake. PMID- 11929917 TI - Taste responses of neurons of the hamster solitary nucleus are enhanced by lateral hypothalamic stimulation. AB - Gustatory responses in the brain stem are modifiable by several physiological factors, including blood insulin and glucose, intraduodenal lipids, gastric distension, and learning, although the neural substrates for these modulatory effects are not known. Stimulation of the lateral hypothalamus (LH) produces increases in food intake and alterations in taste preference behavior, whereas damage to this area has opposite effects. In the present study, we investigated the effects of LH stimulation on the neural activity of taste-responsive cells in the nucleus of the solitary tract (NST) of the hamster. Bipolar stimulating electrodes were bilaterally implanted in the LH, and the responses of 99 neurons in the NST, which were first characterized for their taste sensitivities, were tested for their response to both ipsilateral and contralateral LH stimulation. Half of the taste-responsive cells in the NST (49/99) were modulated by LH stimulation. Contralateral stimulation was more often effective (41 cells) than ipsilateral (13 cells) and always excitatory; 10 cells were excited bilaterally. Six cells were inhibited by ipsilateral stimulation. A subset of these cells (n = 13) was examined for the effects of microinjection of DL-homocysteic acid (DLH), a glutamate receptor agonist, into the LH. The effects of electrical stimulation were completely mimicked by DLH, indicating that cell somata in and around the LH are responsible for these effects. Other cells (n = 14) were tested for the effects of electrical stimulation of the LH on the responses to stimulation of the tongue with 0.032 M sucrose, NaCl, and quinine hydrochloride, and 0.0032 M citric acid. Responses to taste stimuli were more than doubled by the excitatory influence of the LH. These data show that the LH, in addition to its role in feeding and metabolism, exerts descending control over the processing of gustatory information through the brain stem. PMID- 11929918 TI - GABAergic and glutamatergic modulation of spontaneous and motor-cortex-evoked complex spike activity. AB - Olivocerebellar activity is organized such that synchronous complex spikes occur primarily among Purkinje cells located within the same parasagittally oriented strip of cortex. Previous findings have shown that this synchrony distribution is modulated by the release of GABA and glutamate within the inferior olive, which probably act by controlling the efficacy of the electrotonic coupling between olivary neurons. The relative strengths of these two neurotransmitters in modulating the patterns of synchrony were compared by obtaining multiple electrode recordings of spontaneous crus 2a complex spike activity during intraolivary injection of solutions containing a GABA(A) (picrotoxin) and/or AMPA [1,2,3,4-tetrahydro-6-nitro-2,3-dioxo-benzo[f]quinoxaline-7-sulfonamide disodium (NBQX)] receptor antagonist. Injection of either antagonist led to increased synchrony between cells located within the same parasagittally oriented approximately 250-microm-wide cortical strip. Picrotoxin also increased complex spike synchrony among cells located in different cortical strips, leading to a less prominent banding pattern, whereas injections of NBQX tended to decrease complex spike synchrony among such cells, enhancing the banding pattern. The relative strength of these two classes of olivary afferents was assessed by first injecting one of the antagonists alone and then in combination with the other. The enhanced banding pattern of complex spike synchrony following injection of NBQX alone remained during the subsequent combined injection of both antagonists. Furthermore, the widespread synchronization of complex spike activity following injection of picrotoxin alone was partially or completely reversed by combined injection of picrotoxin and NBQX. Changes in the climbing fiber reflex induced by the intraolivary injections paralleled the changes observed for spontaneous complex spike activity, indicating that the effects of picrotoxin and NBQX on the synchrony distribution reflect changes in the pattern of effective coupling of inferior olivary neurons and demonstrating that synchronous complex spike activity does not require simultaneous excitatory input to olivary cells. Finally the pattern of synchrony during motor cortical stimulation was examined. It was found that the patterns of synchrony for motor-cortex-evoked complex spike activity were similar to those of spontaneous activity, indicating an important role for electrotonic coupling in determining the response of the olivocerebellar system to afferent input. Moreover, intraolivary injections of picrotoxin increased the spatial distribution of the evoked response. In sum, the results provide evidence for the hypothesis that electrotonic coupling of inferior olivary neurons via gap junctions is the mechanism underlying complex spike synchrony and that this coupling plays an important role in determining the responses of the olivocerebellar system to synaptic input. PMID- 11929919 TI - Subthreshold oscillations induced by spinal nerve injury in dissociated muscle and cutaneous afferents of mouse DRG. AB - Whole cell patch-clamp recordings were obtained from dissociated mouse lumbar dorsal root ganglion (DRG) neurons. Recordings were made from control neurons and neurons axotomized by transection of the corresponding spinal nerve 1-2 days prior to dissociation. Medium to large muscle and cutaneous afferent neurons were identified by retrograde transport of True Blue or Fluoro-Gold injected into the corresponding peripheral tissue. Action potentials were classified as non inflected spikes (A(0)) and inflected spikes (A(inf)). High-frequency, low amplitude subthreshold membrane potential oscillations were observed in 8% of control A(0) neurons, but their incidence increased to 31% in the nerve injury group. Fifty percent of axotomized muscle afferent A(0) cells displayed oscillations, while 26% of axotomized cutaneous afferents exhibited oscillations. Lower-frequency oscillations were also observed in a small fraction (4%) of A(inf) neurons on strong depolarization. Their numbers were increased after the nerve injury, but the difference was not statistically significant. The oscillations often triggered burst firing in distinct patterns of action potential activity. These results indicate that injury-induced membrane oscillations of DRG neurons, previously observed in whole DRG of rats, are present in dissociated DRG neurons of the adult mouse. Moreover, these observations indicate that both muscle and cutaneous afferents in the A(beta) size range give rise to injury-induced membrane oscillations, with muscle afferents being more prone to develop oscillations. PMID- 11929920 TI - Mechanisms involved in persistent facilitation of neuromuscular synapses in aplysia. AB - Synaptic plasticity can last from a fraction of a second to weeks depending on how it was induced. The mechanisms that underlie short-, intermediate-, and long term plasticity have been intensively studied at central synapses of both vertebrates and invertebrates; however, peripheral plasticity has not received as much attention. In this study, we investigated the mechanisms that contribute to a persistent form of plasticity at neuromuscular synapses in buccal muscle I3a of Aplysia. These synapses are reversibly facilitated by the small cardioactive peptide (SCP), a peptide cotransmitter that is intrinsic to the motor neurons, and persistently facilitated by serotonin (5HT) released from modulatory neurons that are extrinsic to the motor circuit. Many of the short-term effects of 5HT and SCP are mediated by the cAMP pathway, but little is known about the mechanisms that underlie persistent modulation. We were able to eliminate several possible mechanisms. One of these was the possibility that the apparent reversal of SCP's effects was due to desensitization of the SCP receptor. Superfusion for longer periods or with higher concentrations of SCP indicate that the SCP receptors do not desensitize. We also determined that new protein synthesis is not required for the persistent facilitation of EJPs. Another possibility was that 5HT was taken up and slowly re-released. Our results suggest that this mechanism is also unlikely. Activation of the cAMP pathway does not appear to mediate persistent effects; however, 5HT as well as SCP does cause persistent increases in cAMP levels that can prime I3a synapses and increase the effectiveness of activators of the cAMP pathway. Instead, the persistent effects of 5HT are mimicked by phorbol, suggesting that protein kinase C or an Aplysia homologue of unc13 may mediate these effects. These results, in combination with results from experiments on the sensory neurons that contribute to withdrawal reflexes in Aplysia, suggest that the mechanisms for intermediate- and long-term facilitation may reside in all of the synapses involved in the sensory to motor response reflex. PMID- 11929921 TI - Regulation of firing response gain by calcium-dependent mechanisms in vestibular nucleus neurons. AB - Behavioral reflexes can be modified by experience via mechanisms that are largely unknown. Within the circuitry for the vestibuloocular reflex (VOR), neurons in the medial vestibular nucleus (MVN) show adaptive changes in firing rate responses that are correlated with VOR gain (the ratio of evoked eye velocity to input head velocity). Although changes in synaptic strength are typically assumed to underlie gain changes in the VOR, modulation of intrinsic ion channels that dictate firing could also play a role. Little is known, however, about how ion channel function or regulation contributes to firing responses in MVN neurons. This study examined contributions of calcium-dependent currents to firing responses in MVN neurons recorded with whole cell patch electrodes in rodent brain stem slices. Firing responses were remarkably linear over a wide range of firing rates and showed modest spike frequency adaptation. Firing response gain, the ratio of evoked firing rate to input current, was reduced by increasing extracellular calcium and increased either by lowering extracellular calcium or with antagonists to SK- and BK-type calcium-dependent potassium channels and N- and T-type calcium channels. Blockade of SK channels occluded gain increases via N-type calcium channels, while blocking BK channels occluded gain increases via presumed T-type calcium channels, indicating specific coupling of potassium channels and their calcium sources. Selective inhibition of Ca(2+)/calmodulin dependent kinase II and broad-spectrum inhibition of phosphatases modulated gain via BK-dependent pathways, indicating that firing responses are tightly regulated. Modulation of firing response gain by phosphorylation provides an attractive mechanism for adaptive control of VOR gain. PMID- 11929922 TI - Sensitization to mechanical stimulation by inflammatory mediators and by mild burn in canine visceral nociceptors in vitro. AB - Hyperalgesia to mechanical stimulation and heat is commonly observed in inflamed conditions. Although sensitization to heat is well documented and its mechanism has also been well studied, it remains unclear whether and how nociceptors are sensitized to mechanical stimulation. Therefore we conducted in vitro investigation of which inflammatory mediators (bradykinin, histamine, prostaglandin E2, and protons) sensitize nociceptors to suprathreshold mechanical stimulation and at what concentrations. In addition, we studied the effects of possible second messengers for these mediators downstream of the receptors and also the effects of mild burn. Single polymodal receptor activities were recorded in canine testis-spermatic nerve preparations excised from deeply anesthetized dogs. Mechanical stimulation was applied to the identified receptive field for 10 s with a servo-controlled mechanical stimulator. Bradykinin at 0.001 microM induced neither excitation nor facilitation of the mechanical response; however, it facilitated the mechanical response at 0.01 microM and higher, levels at which significant excitation was also induced by bradykinin alone. Histamine excited the nociceptor and sensitized it to mechanical stimulation at 10 microM and higher. PG E(2) also sensitized the mechanical response, but starting at 1 microM, without inducing excitation by itself. The effects of two possible intracellular messengers for these mediators were studied using forskolin (10 microM), which increases intracellular cAMP, and a protein-kinase-C-stimulating phorbol ester, phorbol 12,13-dibutyrate (0.1 microM). Both substances reversibly facilitated the mechanical response of testicular polymodal receptors. In contrast, low-pH solution (pH: 6.6-4.5) seldom induced excitation and failed to facilitate the mechanical response. After 55 degrees C, 30-s heat stimulation, testicular polymodal receptors were sensitized to mechanical stimulation. These results demonstrated that inflammatory mediators and burn sensitized nociceptor responses to mechanical stimulation and provide support for the idea that peripheral nociceptor sensitization is a mechanism involved in hyperalgesia to mechanical stimulation in inflamed tissues. PMID- 11929923 TI - Characterization and comparison of the NR3A subunit of the NMDA receptor in recombinant systems and primary cortical neurons. AB - Recently, we cloned and began to characterize a new N-methyl-D-aspartate receptor (NMDAR) subunit, NR3A. Here we extend our earlier findings by showing that recombinantly expressed NR3A in COS cells is biochemically associated with both NR1 and NR2 subunits. In the oocyte or HEK 293 cell expression systems, co injection of NR3A with NR1/NR2 subunits acts in a dominant-interfering manner, resulting in a decrease in NMDAR unitary conductance, decrease in Ca(2+) permeability, decrease in Mg(2+) sensitivity, and slight increase in mean open time compared with NR1/NR2 channels. The smaller unitary conductance channel has also been observed in primary cortical neurons cultured from wild-type rodent on postnatal day 8 (P8) and similarly found to be relatively insensitive to Mg(2+) block. Consistent with these findings, whole cell NMDA-evoked currents are larger in NR3A-deficient mice compared with wild-type mice, and this effect follows a developmental pattern similar to that of NR3A protein expression on Western blots, with peak expression at P8. Finally, a new longer splice variant of NR3A has been cloned and found to be expressed in rodent cortical neurons by single cell RT-PCR and in situ hybridization. PMID- 11929924 TI - Comparison of human ocular torsion patterns during natural and galvanic vestibular stimulation. AB - Galvanic vestibular stimulation (GVS) is reported to induce interindividually variable tonic ocular torsion (OT) and superimposed torsional nystagmus. It has been proposed that the tonic component results from the activation of otolith afferents. We tested our hypothesis that both the tonic and the phasic OT are mainly due to semicircular canal (SCC) stimulation by examining whether the OT patterns elicited by GVS can be reproduced by pure SCC stimulations. Using videooculography we measured the OT of six healthy subjects while two different stimuli with a duration of 20 s were applied: 1) transmastoidal GVS steps of 2 mA with the head in a pitched nose-down position and 2) angular head rotations around a combined roll-yaw axis parallel to the gravity vector with the head in the same position. The stimulation profile was individually scaled to match the nystagmus properties from GVS and consisted of a sustained velocity step of 4-12 degrees /s on which a velocity ramp of 0.67-2 degrees /s(2) was superimposed. Since blinks were reported to induce transient torsional eye movements, the subjects were also asked to blink once 10 s after stimulus onset. Analysis of torsional eye movements under both conditions revealed no significant differences. Thus we conclude that both the tonic and the phasic OT responses to GVS can be reproduced by pure rotational stimulations and that the OT-related effects of GVS on SCC afferents are similar to natural stimulations at small amplitudes. PMID- 11929925 TI - Bilateral soleus H-reflexes in humans elicited by simultaneous trains of stimuli: symmetry, variability, and covariance. AB - Experiments using electrical and mechanical activation of spinal reflexes have contributed important results toward the understanding of neuronal and synaptic dynamics involved in spinal neural circuits as well as their response to different inputs. In this work, data obtained from the simultaneous stimulation of both legs are analyzed to provide information on the degree of symmetry of the respective spinal reflex circuits and on the characteristics of reflex variability. H-reflexes recorded from relaxed muscles show a frequency-dependent amplitude depression when elicited by a train of stimuli. This effect has been attributed to homosynaptic depression. Soleus H-reflexes were recorded in response to trains of simultaneous stimuli applied to both legs in right-handed subjects that were sitting in a relaxed state. The first objective was to verify the existence of asymmetries in H-reflex parameters obtained from the two legs. We measured the mean, variance, and coefficient of variation of the depressed H reflex amplitudes and the time constant of decay toward the depressed plateau. The second objective was the analysis of the time correlation of subsequent H reflex amplitudes in a long train of responses recorded from a given leg. The statistical dependence of H-reflex amplitudes in the long trains recorded from both legs was also investigated. Data obtained from preliminary experiments showed that there was no effect of a given stimulus on the contralateral leg applied simultaneously or 1 s before, therefore validating the simultaneous stimulation paradigm. Paired t-tests indicated that several parameters measured bilaterally from soleus H-reflex trains of right-handed subjects were not statistically different in the overall, although individually there were statistically significant asymmetries, toward either the right or left leg. Sequences of H-reflex amplitudes, as measured by the auto-covariance, were either white or had a memory ranging from 2 up to 50 s. This indicates that the random fluctuations in presynaptic inhibition and/or postsynaptic inputs to motoneurons may have either fast or slow time courses. The average auto-covariance sequences of the right and left legs, computed from all subjects, were practically superposable. The cross-covariance between the bilateral H-reflex amplitudes showed a statistically significant peak at zero lag in some experiments, suggesting a correlation between the synaptic inputs to the Ia-motoneuron systems of the soleus muscles of both legs. PMID- 11929926 TI - Single-neuron analysis of human thalamus in patients with intention tremor and other clinical signs of cerebellar disease. AB - Tremor that occurs as a result of a cerebellar lesion, cerebellar tremor, is characteristically an intention tremor. Thalamic activity may be related to cerebellar tremor because transmission of some cerebellar efferent signals occurs via the thalamus and cortex to the periphery. We have now studied thalamic neuronal activity in a cerebellar relay nucleus (ventral intermediate-Vim) and a pallidal relay nucleus (ventralis oral posterior-Vop) during thalamotomy in patients with intention tremor and other clinical signs of cerebellar disease (tremor patients). The activity of single neurons and the simultaneous electromyographic (EMG) activity of the contralateral upper extremity in tremor patients performing a pointing task were analyzed by spectral cross-correlation analysis. EMG spectra during intention tremor often showed peaks of activity in the tremor-frequency range (1.9-5.8 Hz). There were significant differences in thalamic neuronal activity between tremor patients and controls. Neurons in Vim and Vop had significantly lower firing rates in tremor patients than in patients undergoing thalamic surgery for pain (pain controls). Other studies have shown that inputs to Vim from the cerebellum are transmitted through excitatory connections. Therefore the present results suggest that tremor in these tremor patients is associated with deafferentation of the thalamus from cerebellar efferent pathways. The thalamic X EMG cross-correlation functions were studied for cells located in Vim and Vop. Neuronal and EMG activity were as likely to be significantly correlated for cells in Vim as for those in Vop. Cells in Vim were more likely to have a phase lag relative to EMG than were cells in Vop. In monkeys, cells in the cerebellar relay nucleus of the thalamus, corresponding to Vim, are reported to lead movement during active oscillations at the wrist. In view of these monkey studies, the present results suggest that cells in Vim are deafferented and have a phase lag relative to tremor that is not found in normal active oscillations. The difference in phase of thalamic spike X EMG activity between Vim and Vop may contribute to tremor because lesions of pallidum or Vop are reported to relieve cerebellar tremor. PMID- 11929927 TI - Vagal mechanoreceptors and chemoreceptors in mouse stomach and esophagus. AB - We used a novel in vitro mouse vagus-gastro-esophageal preparation to study the properties of peripheral vagal afferent endings. We found two types of mechanoreceptive fiber, mucosal receptors and tension receptors. These were distinguished by their sensitivity to mucosal stroking with von Frey hairs and circular tension applied via a claw-cantilever system. A comparison was made with gastro-esophageal afferents found in a similar preparation of ferret tissue. Responses of mouse tension receptors to circular tension were significantly greater than ferret tension and tension/mucosal receptors. Similarly the responses of mouse mucosal receptors to mucosal stroking were significantly greater than ferret mucosal and tension/mucosal receptors. Forty-seven percent of mouse mucosal receptors and 50% of tension receptors responded to one or more drugs or chemical stimuli applied to the receptive field. These included alpha,beta-methylene ATP (10(-6) to 10(-3) M), 5-hydroxytryptamine (10(-6) to 10( 3) M), and hydrochloric acid (10(-2) to 10(-1) M). Drug responses were concentration dependent. One hundred percent of mucosal receptors and 61% of tension receptors tested responded to bile (1:8 to 1:1 dilution). A third type of fiber was recruited by bile. These fibers were mechanically insensitive and silent prior to bile exposure. In conclusion, we have shown three types of gastro esophageal vagal afferent fibers in the mouse: mucosal mechanoreceptors, tension receptors, and specific chemoreceptors activated by bile. PMID- 11929928 TI - Responses of spectrally selective cells in macaque area V2 to wavelengths and colors. AB - We have recorded from wavelength-selective cells in macaque monkey visual area V2, interposed between areas V1 and V4 of the color-specialized pathway, to learn whether their responses correlate with perceived colors or are determined by the wavelength composition of light reflected from their receptive fields. All the cells we recorded from were unselective for the orientation and direction of motion of the stimulus, and all were histologically identified to be in the thin cytochrome oxidase stripes. Using multi-colored "Mondrian" scenes of the appropriate spatial configuration, areas of different color were placed in the receptive field of each cell and the entire scene illuminated by three projectors, passing long-, middle-, and short-wave light, respectively, in various combinations. Our results show that wavelength-selective cells in V2 respond to an area of any color depending on whether or not it reflects a sufficient amount of light of their preferred wavelength. In addition, the responses of a third of the cells tested were also influenced by the wavelength composition of their immediate surrounds, thus signaling the result of a local spatial comparison with respect to the amount of their preferred wavelength present. The responses of all also depended on the sequence with which their receptive fields were illuminated with light of the three different wavebands: cells were activated when there was an increase (and inhibited when there was a decrease) in the amount of their preferred wavelength with respect to the other two; the temporal route taken was therefore a determining factor, and, depending on it, cells would either respond or not to a particular combination of wavelengths. We conclude that although spatiotemporal wavelength comparisons are taking place in the color-specialized subdivisions of area V2, the determination of complete color-constant behavior at the neuronal level requires further processing, in other areas. PMID- 11929929 TI - Directional asymmetry of neurons in cortical areas MT and MST projecting to the NOT-DTN in macaques. AB - The cortical projection to the subcortical pathway underlying the optokinetic reflex was studied using antidromic electrical stimulation in the midbrain structures nucleus of the optic tract and dorsal terminal nucleus of the accessory optic system (NOT-DTN) while simultaneously recording from cortical neurons in the superior temporal sulcus (STS) of macaque monkeys. Projection neurons were found in all subregions of the middle temporal area (MT) as well as in the medial superior temporal area (MST). Antidromic latencies ranged from 0.9 to 6 ms with a median of 1.8 ms. There was a strong bias in the population of cortical neurons projecting to the NOT-DTN for ipsiversive stimulus movement (towards the recording side), whereas in the population of cortical neurons not projecting to the NOT-DTN a more or less equal distribution of stimulus directions was evident. Our data indicate that there is no special area in the posterior STS coding for ipsiversive horizontal stimulus movement. Instead, a specific selection of cortical neurons from areas MT and MST forms the projection to the NOT-DTN and as a subpopulation has the same directional bias as their subcortical target neurons. These findings are discussed in relation to the functional grouping of cortical output as an organizational principle for specific motor responses. PMID- 11929930 TI - Neuromodulatory role of serotonin in the ferret thalamus. AB - Serotonergic fibers broadly innervate the thalamus and may influence the sleep wake cycle, attention, and other processes through modulation of neurons in this structure. However, the actions of serotonin in the dorsal thalamus have been investigated in detail only in the dorsal lateral geniculate nucleus. In the present study, we examined the action of serotonin in several different regions of the ferret dorsal thalamus, including the associative nuclei, using the in vitro slice preparation and intracellular recording techniques. In nearly all nuclei examined, the predominant action of serotonin was one of hyperpolarization and inhibition of the tonic firing mode. The magnitude of the hyperpolarizing response decreased with age and varied greatly across and somewhat within nuclei maintaining the following relationship (in descending order of magnitude): lateral posterior, lateral dorsal, pulvinar, mediodorsal, center median, anteroventral, central lateral, ventral basal, and medial geniculate. This hyperpolarization is elicited through two mechanisms: one direct and the other via local interneurons. The direct action occurs through an increase in potassium conductance mediated through the 5-HT(1A) receptor. This conclusion is supported by the findings that it persists in the presence of tetrodotoxin and block of GABAergic synaptic transmission, the reversal potential shifts in a Nernstian fashion with changes in extracellular potassium concentration, and the response is antagonized by the 5-HT(1A) antagonist WAY100635 and mimicked by the application of the 5-HT(1A)-selective agonist 8-OH DPAT. The second mechanism by which 5-HT evoked a hyperpolarization was through the activation of local interneurons. In slices in which GABA receptors were not blocked, 5-HT application increased the frequency and amplitude of spontaneous inhibitory postsynaptic potentials (IPSPs) occurring in thalamocortical neurons. Application of 5-HT to physiologically or morphologically identified interneurons evoked a prolonged suprathreshold depolarization. Our results suggest that serotonergic inputs act differentially across the thalamus in a complex manner involving direct and indirect mechanisms. It appears that 5-HT has a greater direct postsynaptic inhibitory influence in the posterior, medial, and intralaminar nuclei than in the primary sensory nuclei. PMID- 11929931 TI - Coherent electrical activity between vibrissa sensory areas of cerebellum and neocortex is enhanced during free whisking. AB - We tested if coherent signaling between the sensory vibrissa areas of cerebellum and neocortex in rats was enhanced as they whisked in air. Whisking was accompanied by 5- to 15-Hz oscillations in the mystatial electromyogram, a measure of vibrissa position, and by 5- to 20-Hz oscillations in the differentially recorded local field potential (nablaLFP) within the vibrissa area of cerebellum and within the nablaLFP of primary sensory cortex. We observed that only 10% of the activity in either cerebellum or sensory neocortex was significantly phase-locked to rhythmic motion of the vibrissae; the extent of this modulation is in agreement with the results from previous single-unit measurements in sensory neocortex. In addition, we found that 40% of the activity in the vibrissa areas of cerebellum and neocortex was significantly coherent during periods of whisking. The relatively high level of coherence between these two brain areas, in comparison with their relatively low coherence with whisking per se, implies that the vibrissa areas of cerebellum and neocortex communicate in a manner that is incommensurate with whisking. To the extent that the vibrissa areas of cerebellum and neocortex communicate over the same frequency band as that used by whisking, these areas must multiplex electrical activity that is internal to the brain with activity that is that phase-locked to vibrissa sensory input. PMID- 11929932 TI - HSV-1 helper virus 5dl1.2 suppresses sodium currents in amplicon-transduced neurons. AB - The Herpes Simplex Virus-1 (HSV-1) amplicon system is one of several viral-based strategies currently being developed for gene delivery into mammalian neurons for experimental or therapeutic purposes. Amplicon-containing viruses contain no HSV 1 genes and are amplified in titer relative to the helper viruses used to package them. In this way, they are designed to have a minimal impact on the physiology of transduced neurons. We show here, however, that amplicon preparations made using the 5dl1.2 helper virus selectively suppress sodium currents in cultured neurons by approximately 80% within 2 days of transduction and reduce average spike frequency in response to depolarization from 23 +/- 4 to 0.4 +/- 0.4 Hz. We observe similar suppression of Na(+) currents in cells treated with the 5dl1.2 helper virus alone, indicating that the helper virus retains the ability of wild type HSV-1 to inhibit these currents potently. Staining amplicon-transduced neurons with anti-HSV antibodies, we find that 80% of the neurons express viral proteins, indicating that helper virus typically co-infects these cells. We conclude that Na(+) current suppression by the amplicon preparation results from the preferential coinfection of transduced neurons by the 5dl1.2 helper virus. PMID- 11929933 TI - Distribution of eye- and arm-movement-related neuronal activity in the SEF and in the SMA and Pre-SMA of monkeys. AB - We analyzed neuronal activity in the supplementary eye field (SEF), supplementary motor area (SMA), and presupplementary motor area (pre-SMA) during the performance of three motor tasks: capturing a visual target with a saccade, reaching one arm to a target while gazing at a visual fixation point, or capturing a target with a saccade and arm-reach together. Our data demonstrated that each area was involved in controlling the arm and eye movements in a different manner. Saccade-related neurons were found mainly in the SEF. In contrast, arm-movement-related neurons were found primarily in the SMA and pre SMA. In addition, we found that the activity of both arm-movement- and saccade related neurons differed depending on the presence or absence of an accompanying saccade or arm movement. Such context dependency was found in all three areas. We also discovered that activity preceding eye or arm movement alone, and eye and arm movement combined, appeared more often in the pre-SMA and SEF, suggesting their involvement in effector-independent aspects of motor behavior. Subsequent analysis revealed that the laterality of arm representation differed in the three areas: it was predominantly contralateral in the SMA but largely bilateral in the pre-SMA and SEF. PMID- 11929934 TI - D1/D5 dopamine receptors stimulate intracellular calcium release in primary cultures of neocortical and hippocampal neurons. AB - D1/D5 dopamine receptors in basal ganglia, hippocampus, and cerebral cortex modulate motor, reward, and cognitive behavior. Previous work with recombinant proteins revealed that in cells primed with heterologous G(q/11)-coupled G protein-coupled receptor (GPCR) agonists, the typically G(s)-linked D1/D5 receptors can stimulate robust release of calcium from internal stores when coexpressed with calcyon. To learn more about the intracellular signaling mechanisms underlying these D1/D5 receptor regulated behaviors, we explored the possibility that endogenous receptors stimulate internal release of calcium in neurons. We have identified a population of neurons in primary cultures of hippocampus and neocortex that respond to D1/D5 dopamine receptor agonists with a marked increase in intracellular calcium (Ca) levels. The D1/D5 receptor stimulated responses occurred in the absence of extracellular Ca(2+) indicating the rises in Ca involve release from internal stores. In addition, the responses were blocked by D1/D5 receptor antagonists. Further, the D1/D5 agonist-evoked responses were state dependent, requiring priming with agonists of G(q/11) coupled glutamate, serotonin, muscarinic, and adrenergic receptors or with high external K(+) solution. In contrast, D1/D5 receptor agonist-evoked Ca(2+) responses were not detected in neurons derived from striatum. However, D1/D5 agonists elevated cAMP levels in striatal cultures as effectively as in neocortical and hippocampal cultures. Further, neither forskolin nor 8-Br-cAMP stimulation following priming was able to mimic the D1/D5 agonist-evoked Ca(2+) response in neocortical neurons indicating that increased cAMP levels are not sufficient to stimulate Ca release. Our data suggest that D1-like dopamine receptors likely modulate neocortical and hippocampal neuronal excitability and synaptic function via Ca(2+) as well as cAMP-dependent signaling. PMID- 11929935 TI - Computational study on monkey VOR adaptation and smooth pursuit based on the parallel control-pathway theory. AB - Much controversy remains about the site of learning and memory for vestibuloocular reflex (VOR) adaptation in spite of numerous previous studies. One possible explanation for VOR adaptation is the flocculus hypothesis, which assumes that this adaptation is caused by synaptic plasticity in the cerebellar cortex. Another hypothesis is the model proposed by Lisberger that assumes that the learning that occurs in both the cerebellar cortex and the vestibular nucleus is necessary for VOR adaptation. Lisberger's model is characterized by a strong positive feedback loop carrying eye velocity information from the vestibular nucleus to the cerebellar cortex. This structure contributes to the maintenance of a smooth pursuit driving command with zero retinal slip during the steady state phase of smooth pursuit with gain 1 or during the target blink condition. Here, we propose an alternative hypothesis that suggests that the pursuit driving command is maintained in the medial superior temporal (MST) area based on MST firing data during target blink and during ocular following blank, and as a consequence, we assume a much smaller gain for the positive feedback from the vestibular nucleus to the cerebellar cortex. This hypothesis is equivalent to assuming that there are two parallel neural pathways for controlling VOR and smooth pursuit: a main pathway of the semicircular canals to the vestibular nucleus for VOR, and a main pathway of the MST-dorsolateral pontine nuclei (DLPN) flocculus/ventral paraflocculus to the vestibular nucleus for smooth pursuit. First, we theoretically demonstrate that this parallel control-pathway theory can reproduce the various firing patterns of horizontal gaze velocity Purkinje cells in the flocculus/ventral paraflocculus dependent on VOR in the dark, smooth pursuit, and VOR cancellation as reported in Miles et al. at least equally as well as the gaze velocity theory, which is the basic framework of Lisberger's model. Second, computer simulations based on our hypothesis can stably reproduce neural firing data as well as behavioral data obtained in smooth pursuit, VOR cancellation, and VOR adaptation, even if only plasticity in the cerebellar cortex is assumed. Furthermore, our computer simulation model can reproduce VOR adaptation automatically based on a heterosynaptic interaction model between parallel fiber inputs and climbing fiber inputs. Our results indicate that different assumptions about the site of pursuit driving command maintenance computationally lead to different conclusions about where the learning for VOR adaptation occurs. Finally, we propose behavioral and physiological experiments capable of discriminating between these two possibilities for the site of pursuit driving command maintenance and hence for the sites of learning and memory for VOR adaptation. PMID- 11929936 TI - Dopamine excites fast-spiking interneurons in the striatum. AB - The striatum is the main recipient of dopaminergic innervation. Striatal projection neurons are controlled by cholinergic and GABAergic interneurons. The effects of dopamine on projection neurons and cholinergic interneurons have been described. Its action on GABAergic interneurons, however, is still unknown. We studied the effects of dopamine on fast-spiking (FS) GABAergic interneurons in vitro, with intracellular recordings. Bath application of dopamine elicited a depolarization accompanied by an increase in membrane input resistance (an effect that persisted in the presence of tetrodotoxin) and action-potential discharge. These effects were mimicked by the D1-like dopamine receptor agonist SKF38393 but not by the D2-like agonist quinpirole. Evoked corticostriatal glutamatergic synaptic currents were not affected by dopamine. Conversely, GABAergic currents evoked by intrastriatal stimulation were reversibly depressed by dopamine and D2 like, but not D1-like, agonists. Cocaine elicited effects similar to those of dopamine on membrane potential and synaptic currents. These results show that endogenous dopamine exerts a dual excitatory action on FS interneurons, by directly depolarizing them (through D1-like receptors) and by reducing their synaptic inhibition (through presynaptic D2-like receptors). PMID- 11929937 TI - Electrical stimulation of a denervated muscle promotes selective reinnervation by native over foreign motoneurons. AB - The effect of electrical stimulation of the denervated posterior cricoarytenoid (PCA) muscle on its subsequent reinnervation was explored in the canine. Eight animals were implanted with a planar array of 36 electrodes for chronic stimulation and recording of spontaneous and evoked electromyographic (EMG) potentials across the entire fan-shaped surface of a muscle pair. Normative EMG data were recorded from each electrode site before unilateral nerve section, and from the innervated partner after nerve section. After randomizing the animals to experimental and control groups, the right recurrent laryngeal nerve innervating the PCA abductor muscle and its adductor antagonists was sectioned and reanastomosed. The PCA muscle in four experimental animals was continuously stimulated during the 11-mo experiment, using a 1-s, 30-pps, biphasic pulse train composed of 1-ms pulses 2-6 mA in amplitude and repeated every 10 s. The remaining four animals served as nonstimulated controls. Appropriate reinnervation by native inspiratory motoneurons was indexed behaviorally by the magnitude of vocal fold opening and electromyographically by the potential across all electrode sites. Inappropriate reinnervation by foreign adductor motoneurons was quantitated by recording EMG potentials evoked reflexly by stimulation of sensory afferents of the laryngeal mucosa. All four experimental animals showed a greater level of correct PCA muscle reinnervation (P < 0.0064) and a lesser level of incorrect reinnervation (P < 0.0084) than the controls. Direct muscle stimulation also appeared to enhance the overall magnitude of reinnervation, but the effect was not as strong (P < 0.113). These findings are consistent with a previous report and suggest that stimulation of a mammalian muscle may profoundly affect its receptivity to reinnervation by a particular motoneuron type. PMID- 11929938 TI - Common drive in motor units of a synergistic muscle pair. AB - The interaction among the motor units of the extensor carpi radialis longus (ECRL) and the extensor carpi ulnaris (ECU) muscles in man was studied during wrist extensions in which the two muscles acted as synergists. Intramuscular recordings were obtained using special quadrifilar needle electrodes. Isometric wrist extensions at 20-30% of the maximal effort were studied. The electromyographic (EMG) signals were decomposed into the individual motor-unit action potential trains comprising the signal. The interaction among motor units were characterized by the estimated time-varying mean firing rate and the cross correlation between the time-varying mean firing rates of pairs of motor units. Pairs of motor units within each muscle as well as pairs of motor units across the muscles were considered. In-phase common fluctuations, termed common drive, were observed in the mean firing rates of motor units within each muscle, consistent with earlier work on other muscles. Common fluctuations were also observed between the firing rates of ECU and ECRL motor units albeit with a variable phase shift. The existence of common drive across synergistic muscles was interpreted as implying that the CNS considers the muscles as a functional unit when they act as synergists. PMID- 11929939 TI - Transient analgesia evoked by noxious stimulus offset. AB - Pain has long been thought to wax and wane in relative proportion to fluctuations in the intensity of noxious stimuli. Dynamic aspects of nociceptive processing, however, remain poorly characterized. Here we show that small decreases (+/-1-3 degrees C) in noxious stimulus temperatures (47-50 degrees C) evoked changes in perceived pain intensity that were as much as 271% greater than those of equal magnitude increases. These decreases in perceived pain intensity were sufficiently large to be indistinguishable from those evoked by 15 degrees C decreases to clearly innocuous levels. Furthermore, decreases in pain ratings following noxious stimulus offset were significantly greater than those occurring during adaptation to constant temperature stimuli. Together, these findings indicate that an analgesic mechanism is activated during noxious stimulus offset. This analgesic phenomenon may serve as a temporal contrast enhancement mechanism to amplify awareness of stimulus offset and to reinforce escape behaviors. Disruption of this mechanism may contribute importantly to chronic pain. PMID- 11929940 TI - Is TIG1 a new tumor suppressor in prostate cancer? PMID- 11929941 TI - Disaggregating the effects of race and poverty on breast cancer outcomes. PMID- 11929942 TI - Upstaging tamoxifen? New classes of drugs emerging for breast cancer. PMID- 11929943 TI - SNPs: can genetic variants control cancer susceptibility? PMID- 11929944 TI - Mortality from esophageal cancer in selected countries, 1998. PMID- 11929945 TI - Networks aim to bridge gap between clinical research, medical practice. PMID- 11929946 TI - Use of breast-conserving therapy increases, but variations persist. PMID- 11929948 TI - Tazarotene-induced gene 1 (TIG1) expression in prostate carcinomas and its relationship to tumorigenicity. AB - BACKGROUND: Prostate cancer is the most common noncutaneous male cancer and one of the least understood malignant diseases. Identifying key genetic factors involved in the metastasis of prostate cancer cells is critical. In this study, we used selective subtractive differential gene display to identify a gene whose decreased expression may contribute to the growth and expansion of prostate cancer. METHODS: We used 192 primer pair combinations and polymerase chain reaction technology to identify genes expressed in the benign prostate cell line PNT-2 but not in the malignant prostate cancer cell lines LNCaP, Du-145, PC-3, or PC-3M. The tazarotene-induced gene 1 (TIG1) was chosen for further study. TIG1 expression in normal tissues and cell lines was analyzed by northern blot and in normal and tumor prostate tissue sections by in situ hybridization. The in vitro invasiveness (migration through extracellular matrix) and in vivo tumorigenicity (growth in nude mice) were assessed for the highly malignant PC-3M cell line transfected with TIG1 or control cDNA. All statistical tests were two-sided. RESULTS: TIG1 mRNA was expressed in a variety of normal tissues other than prostate tissue. TIG1 mRNA was detected in all 10 normal human prostate tissues and all 51 benign prostatic hyperplastic tissues analyzed but in only four of 51 malignant prostate tissues analyzed. Compared with vector-transfected cells, transfection of PC-3M cells with TIG1 decreased in vitro invasiveness from 14.7% to 3.7%, (mean difference = 11%; 95% confidence interval [CI] = 9.2% to 12.8%, P<.001) and decreased in vivo tumorigenicity from an average tumor weight of 1.31 g to 0.55 g, (mean difference = 0.76 g; 95% CI = 0.43 to 1.09 g, P<.001). CONCLUSION: TIG1 may be a tumor suppressor gene whose diminished expression is involved in the malignant progression of prostate cancer. PMID- 11929949 TI - Race, socioeconomic status, and breast cancer treatment and survival. AB - BACKGROUND: Previous studies have found that African-American women are more likely than white women to have late-stage breast cancer at diagnosis and shortened survival. However, there is considerable controversy as to whether these differences in diagnosis and survival are attributable to race or socioeconomic status. Our goal was to disentangle the influence of race and socioeconomic status on breast cancer stage, treatment, and survival. METHODS: We linked data from the Metropolitan Detroit Surveillance, Epidemiology, and End Results (SEER)(1) registry to Michigan Medicaid enrollment files and identified 5719 women diagnosed with breast cancer, of whom 593 were insured by Medicaid. We first calculated the unadjusted odds ratios (ORs) associated with race, Medicaid insurance, and poverty for breast cancer stage at diagnosis, breast cancer treatment, and death. We then estimated the ORs of having late-stage breast cancer at diagnosis, breast-conserving surgery, no surgery, and death using logistic regression after controlling for clinical and nonclinical factors. All statistical tests were two-sided. RESULTS: Before controlling for Medicaid enrollment and poverty, African-American women had a higher likelihood than white women of each unfavorable breast cancer outcome. However, after controlling for covariates, African-American women were not statistically significantly different from white women on most outcomes except for surgical choice. African-American women were more likely than white women to have no surgery (adjusted OR = 1.62; 95% confidence interval [CI] = 1.11 to 2.37). Among women who had surgery, African-American women were more likely to have breast-conserving surgery than were white women (adjusted OR = 1.63; 95% CI = 1.33 to 1.98). CONCLUSIONS: The linkage of Medicaid and SEER data provides more in-depth information on low income women than has been available in past studies. In our Metropolitan Detroit study population, race was not statistically significantly associated with unfavorable breast cancer outcomes. However, low socioeconomic status was associated with late-stage breast cancer at diagnosis, type of treatment received, and death. PMID- 11929950 TI - Hormone replacement therapy and the risk of invasive epithelial ovarian cancer in Swedish women. AB - BACKGROUND: Estrogen replacement therapy (ERT), which is mainly used to relieve climacteric symptoms, increases a woman's risk for uterine endometrial cancer and epithelial ovarian cancer (EOC). Estrogens are often combined with progestins in hormone replacement therapy (HRT) to reduce the risk of uterine endometrial cancer. Data on the association between HRT including progestins and EOC risk are limited. This nationwide case-control study examined EOC risk in relation to HRT regimens with sequentially added progestins (HRTsp) and continuously added progestins (HRTcp). METHODS: Between 1993 and 1995, we enrolled 655 histologically verified incident case patients with EOC and 3899 randomly selected population controls, all 50-74 years of age. Data on HRT use were collected through mailed questionnaires. Multivariate-adjusted odds ratios (ORs) and 95% confidence intervals (CIs) were estimated by the use of unconditional logistic regression. RESULTS: Risks of EOC were elevated among ever users as compared with never users of both ERT (OR = 1.43, 95% CI = 1.02 to 2.00) and HRTsp (OR = 1.54, 95% CI = 1.15 to 2.05); risks were elevated for serous, mucinous, and endometrioid subtypes. For all EOC types combined, the greatest risk increases were seen with hormone use exceeding 10 years. Ever use of HRTcp was not associated with increased EOC risk relative to HRTcp never use (OR = 1.02, 95% CI = 0.73 to 1.43). The risk of EOC was elevated among HRTsp ever users as compared with HRTcp ever users (OR = 1.78, 95% CI = 1.05 to 3.01). ORs for EOC after ever use of low-potency estrogens were 1.18 (95% CI = 0.89 to 1.55) for oral and 1.33 (95% CI = 1.03 to 1.72) for vaginal applications, but no relationship was seen between EOC risk and duration of use. CONCLUSION: Ever users of ERT and HRTsp but not HRTcp may be at increased risk of EOC. PMID- 11929951 TI - Modulation of p53, ErbB1, ErbB2, and Raf-1 expression in lung cancer cells by depsipeptide FR901228. AB - BACKGROUND: Histone deacetylases (HDACs) modulate chromatin structure by regulating acetylation of core histone proteins. HDAC inhibitors, such as depsipeptide FR901228 (FK228), induce growth arrest and apoptosis in a variety of human cancer cells by mechanisms that cannot be attributed solely to histone acetylation. This study evaluated the mechanisms by which FK228 mediates apoptosis in non-small-cell lung cancer (NSCLC) cells. METHODS: Proliferation and apoptosis were assessed in a panel of NSCLC cell lines that vary in the expression of the growth-regulating proteins p53, pRb, and K-Ras treated with a clinically relevant dose of FK228 (25 ng/mL). Western blot and immunoprecipitation techniques were used to analyze expression of cell-cycle proteins (cyclin A, cyclin E, p53, and p21), signaling-related proteins (ErbB1, ErbB2, and Raf-1), activity of extracellular signal-regulated kinase 1 and 2 (ERK1/2), binding of mutant p53 and Raf-1 to heat shock protein (Hsp)90, and acetylation of Hsp90. RESULTS: FK228 treatment inhibited growth and induced apoptosis in NSCLC cells expressing wild-type or mutant p53. FK228 treatment led to altered expression of cyclin A, cyclin E, and p21, and to reduced expression of mutant, but not wild-type, p53. FK228-treated cells also were depleted of ErbB1, ErbB2, and Raf-1 proteins, and exhibited lower ERK1/2 activity. FK228 treatment also inhibited the binding of mutant p53 and Raf-1 to Hsp90; this inhibition was associated with acetylation of Hsp90. CONCLUSIONS: FK228 depletes the levels of several oncoproteins that are normally stabilized by binding to Hsp90 in cancer cells. The resulting ability of FK228 to diminish signal transduction via pathways involving Raf-1 and ERK may contribute to the potency and specificity of this novel antitumor agent. PMID- 11929952 TI - Osteopontin identified as lead marker of colon cancer progression, using pooled sample expression profiling. AB - BACKGROUND: New tumor markers and markers of tumor progression are needed for improved staging and for better assessment of treatment of many cancers. Gene expression profiling techniques offer the opportunity to discover such markers. We investigated the feasibility of sample pooling strategy in combination with a novel analysis algorithm to identify markers. METHODS: Total RNA from human colon tumors (n = 60) of multiple stages (adenomas; cancers with modified Astler Collier stages B, C, and D; and liver metastases) were pooled within stages and compared with pooled normal mucosal specimens (n = 10) by using oligonucleotide expression arrays. Genes that showed consistent increases or decreases in their expression through tumor progression were identified. Northern blot analysis was used to validate the findings. All statistical tests were two-sided. RESULTS: More than 300 candidate tumor markers and more than 100 markers of tumor progression were identified. Northern analysis of 11 candidate tumor markers confirmed the gene expression changes. The gene for the secreted integrin-binding protein osteopontin was most consistently differentially expressed in conjunction with tumor progression. Its potential as a progression marker was validated (Spearman's rho = 0.903; P<.001) with northern blot analysis using RNA from an independent set of 10 normal and 43 tumor samples representing all stages. Moreover, a statistically significant correlation between osteopontin protein expression and advancing tumor stage was identified with the use of 303 additional specimens (human cancer = 185, adenomas = 67, and normal mucosal specimens = 51) (Spearman's rho = 0.667; P<.001). CONCLUSIONS: Sample pooling can be a powerful, cost-effective, and rapid means of identifying the most common changes in a gene expression profile. We identified osteopontin as a clinically useful marker of tumor progression by use of gene expression profiling on pooled samples. PMID- 11929954 TI - Re: Effect of age on risk of second primary colorectal cancer. PMID- 11929953 TI - Activation of cancer-specific gene expression by the survivin promoter. AB - BACKGROUND: Survivin, a member of the IAP (inhibitor of apoptosis) gene family, appears to be overexpressed in common cancers but not in corresponding normal adult tissues. To investigate whether the survivin promoter controls cancer cell specific gene expression, we determined whether the survivin gene promoter could regulate reporter gene expression in cancer cell lines and xenografts. METHODS: Survivin protein levels were determined in human and murine cancer cell lines and in normal tissues of adult C57BL/6 mice by Western blot analysis. A reporter construct in which a portion of the survivin gene promoter was used to drive transcription of a human secreted alkaline phosphatase (SEAP) gene was transiently transfected into cancer cells, and promoter activity was extrapolated from SEAP activity. A2780 human ovarian cancer cells were transfected with this construct, and stable transfectants were injected into the intrabursal ovarian space of immunodeficient mice. Tumor growth was monitored, and plasma SEAP levels were used as a measure of survivin promoter activity in vivo. RESULTS: Survivin protein was detected in all cancer cell lines examined but not in most normal adult mouse tissues. After transfection, the survivin promoter was more active in all cancer cell lines than in normal ovarian surface epithelial cells or mouse 3T3 cells. After 0.8 x 10(6) stable transfectant cells were injected into the intrabursal cavity of mouse ovaries, plasma SEAP activity was detected within 24 hours, and the activity increased with time and tumor growth. CONCLUSION: Transfection experiments indicate that survivin protein expression in cancer tissue appears to be regulated, at least in part, transcriptionally. Thus, the survivin promoter may be useful in controlling gene expression in cancer cells. PMID- 11929956 TI - Re: Night shift work, light at night, and risk of breast cancer. PMID- 11929957 TI - Re: Night shift work, light at night, and risk of breast cancer. PMID- 11929958 TI - Re: Night shift work, light at night, and risk of breast cancer. PMID- 11929961 TI - The defective kernel 1 (dek1) gene required for aleurone cell development in the endosperm of maize grains encodes a membrane protein of the calpain gene superfamily. AB - Endosperm of cereal grains is one of the most important renewable resources for food, feed, and industrial raw material. It consists of four triploid cell types, i.e., aleurone, starchy endosperm, transfer cells, and cells of the embryo surrounding region. In maize, the aleurone layer is one cell layer thick and covers most of the perimeter of the endosperm. Specification of maize aleurone cell fate is proposed to occur through activation of the tumor necrosis factor receptor-like receptor kinase CRINKLY4. A second maize gene essential for aleurone cell development is defective kernel 1 (dek1). Here we show that DEK1 shares high homology with animal calpains. The predicted 2,159-aa DEK1 protein has 21 transmembrane regions, an extracellular loop, and a cysteine proteinase domain that shares high homology with domain II of m-calpain from animals. We propose that DEK1 functions to maintain and restrict the aleurone cell fate imposed by CR4 through activation of its cysteine proteinase by contact with the outer endosperm surface. DEK1 seems to be the only member of the calpain superfamily in plants, Arabidopsis DEK1 sharing 70% overall identity with maize DEK1. The expression of dek1 in most plant tissues in maize and Arabidopsis, as well as its presence in a variety of higher plants, including angiosperms and gymnosperms, suggests that DEK1 plays a conserved role in plant signal transduction. PMID- 11929962 TI - Anion-templated self-assembly of tetrahedral cage complexes of cobalt(II) with bridging ligands containing two bidentate pyrazolyl-pyridine binding sites. AB - The bridging ligands L(1) and L(2) contain two N,N-bidentate pyrazolyl-pyridine units linked to a central aromatic spacer unit (1,2-phenyl or 2,3-naphthyl, respectively). Reaction with Ni(II) salts and treatment with the anions tetrafluoroborate or perchlorate result in formation of dinuclear complexes having a 2:3 metal:ligand ratio, with one bridging and two terminal tetradentate ligands. In contrast, reaction of L(1) and L(2) with Co(II) salts, followed by treatment with tetrafluoroborate or perchlorate, results in assembly of cage complexes having a 4:6 metal:ligand ratio; these complexes have a metal ion at each corner of an approximate tetrahedron, and a bis-bidentate bridging ligand spanning each edge. The central cavity is occupied by a tetrahedral counterion that forms multiple hydrogen-bonding interactions with the methylene protons of the bridging ligands. The anionic guest fits tightly into the central cavity of the cage to which it is ideally complementary in terms of shape, size, and charge. Solution NMR experiments show that the central anion acts as a template for cage formation, with a mixture of Co(II) and the appropriate bridging ligand alone giving no assembly into a cage until the tetrahedral anion is added, at which point cage assembly is fast and quantitative. The difference between the structures of the complexes with Ni(II) and Co(II) illustrate how the uncoordinated anions can exert a profound influence on the course of the assembly process. PMID- 11929963 TI - Chemically induced supramolecular reorganization of triblock copolymer assemblies: trapping of intermediate states via a shell-crosslinking methodology. AB - The mechanism of morphological phase transitions was studied for rod-shaped supramolecular assemblies comprised of a poly(acrylic acid)-block-poly(methyl acrylate)-block-polystyrene (PAA(90)-b-PMA(80)-b-PS(100)) triblock copolymer in 33% tetrahydrofuran/water after perturbation by reaction with a positively charged water-soluble carbodiimide. Tetrahydrofuran solvation of the hydrophobic core domain provided the dynamic nature required for the rod-to-sphere phase transition to be complete within 30 min. The intermediate morphologies such as fragmenting rods and pearl-necklace structures were trapped kinetically by the subsequent addition of a diamino crosslinking agent, which underwent covalent crosslinking of the shell layer. Alternatively, shell-crosslinked rod-shaped nanostructures with preserved morphology were obtained by the addition of the crosslinking agent before the addition of the carbodiimide, which allowed for the shell crosslinking to be performed at a faster rate than the morphological reorganization. The formation of robust shell-crosslinked nanostructures provides a methodology by which the morphological evolution processes can be observed, and it allows access to otherwise thermodynamically unstable nanostructures. PMID- 11929964 TI - Pillared honeycomb nanoarchitectures formed on solid surfaces by the self assembly of lipid-packaged one-dimensional Pt complexes. AB - A highly lipophilic polyion complex [Pt(en)(2)][PtCl(2)(en)(2)](1)(4) (en, 1,2 diaminoethane) is prepared from one-dimensional mixed valence Pt(II)/Pt(IV) complex and newly designed chiral amphiphile 1. The powdery sample showed purple color, which is a result of the mixed valence absorption of the linear chlorobridged complex (Pt(II)-Cl-Pt(IV)-Cl-)(n). When the lipid complex is dispersed in dichloromethane, purple-colored dispersion is obtained at 0 degrees C, whereas the color disappears after heating the solution to 21 degrees C. The observed thermochromism is reversible with respect to the temperature changes and is ascribed to the reversible dissociation and reassembly of the self-assembling inorganic wires. Casting of the 0 degrees C-purple dispersion on solid substrates affords honeycomb nanostructures in addition to the nanowires with the width of about 20 nm. The honeycomb patterns seem to be templated by the condensed water droplets that are formed and aligned on the rapidly evaporating dichloromethane solution. On the other hand, more regular honeycomb structures are exclusively obtained by casting the 21 degrees C-colorless solution. These observations indicate that the ordered honeycomb structures are obtainable on solid surfaces by the self-assembly of molecularly dispersed components [Pt(en)(2)](1)(2) and trans-[PtCl(2)(en)(2)](1)(2.) Very interestingly, formation of double-layered honeycomb nanostructure is observed by scanning electron microscopy. The unit hexagons and pillars of the honeycombs are made of nanowires that are hierarchically assembled from the lipid-packaged Pt(II)/Pt(IV) complexes. The surface self-organization of lipophilic inorganic complexes has a potential to fabricate novel nanoarchitectures with conjugated electronic structures. PMID- 11929965 TI - Phase transfer of monosaccharides through noncovalent interactions: selective extraction of glucose by a lipophilic cage receptor. AB - We have previously shown that macrotricyclic host 1a is a powerful receptor for glucopyranosyl units in the nonpolar medium of chloroform. However, the solubility properties of 1a did not permit studies of the extraction of carbohydrates from aqueous solution. This paper describes the synthesis of the new variant 1b, furnished with a highly lipophilic exterior array of 12 benzyloxy substituents. In homogeneous solution, 1b behaves much as 1a, binding n-octyl beta-d-glucoside with K(a) = 720 M(-1) in CD(3)OH/CDCl(3) (8:92). In two-phase experiments, the improved solubility of 1b allows carbohydrate extraction to be observed. Three hexoses (glucose, galactose, and mannose), two pentoses (ribose and xylose), and the two methyl glucosides are all extracted substantially into chloroform from 1 M aqueous solutions. Among the hexoses, 1b shows notable affinity and selectivity for glucose, extracting detectable amounts even from 0.1 M aqueous solutions. PMID- 11929967 TI - Cytosine substituted calix[4]pyrroles: neutral receptors for 5'-guanosine monophosphate. AB - The synthesis and characterization of two cytosine-substituted calix[4]pyrrole conjugates, bearing the appended cytosine attached at either a beta- or meso pyrrolic position, is described. These systems were tested as nucleotide selective carriers and as active components of nucleotide-sensing ion-selective electrodes at pH 6.6. Studies of carrier selectivity were made using a Pressman type model membrane system consisting of an initial pH 6.0 aqueous phase, an intervening dichloromethane barrier containing the calix[4]pyrrole conjugate, and a receiving basic aqueous phase. Good selectivity for the Watson-Crick complementary nucleotide, 5'-guanosine monophosphate (5'-GMP), was seen in the case of the meso-linked conjugate with the relative rates of through-membrane transport being 7.7:4.1:1 for 5'-GMP, 5'-AMP, and 5'-CMP, respectively. By contrast, the beta-substituted conjugate, while showing a selectivity for 5'-GMP that was enhanced relative to unsubstituted calix[4]pyrrole, was found to transport 5'-CMP roughly 4.5 times more quickly than 5'-GMP. Higher selectivities were also found for 5'-CMP when both the beta- and meso-substituted conjugates were incorporated into polyvinyl chloride membranes and tested as ion selective electrodes at pH 6.6, whereas near-equal selectivities were observed for 5'-CMP and 5'-GMP in the case of unsubstituted calix[4]pyrroles. These seemingly disparate results are consistent with a picture wherein the meso-substituted cytosine calix[4]pyrrole conjugate, but not its beta-linked congener, is capable of acting as a ditopic receptor, binding concurrently both the phosphate anion and nucleobase portions of 5'-GMP to the calixpyrrole core and cytosine "tails" of the molecule, respectively, with the effect of this binding being most apparent under the conditions of the transport experiments. PMID- 11929966 TI - A common mutation in the 5,10-methylenetetrahydrofolate reductase gene affects genomic DNA methylation through an interaction with folate status. AB - DNA methylation, an essential epigenetic feature of DNA that modulates gene expression and genomic integrity, is catalyzed by methyltransferases that use the universal methyl donor S-adenosyl-l-methionine. Methylenetetrahydrofolate reductase (MTHFR) catalyzes the synthesis of 5-methyltetrahydrofolate (5 methylTHF), the methyl donor for synthesis of methionine from homocysteine and precursor of S-adenosyl-l-methionine. In the present study we sought to determine the effect of folate status on genomic DNA methylation with an emphasis on the interaction with the common C677T mutation in the MTHFR gene. A liquid chromatography/MS method for the analysis of nucleotide bases was used to assess genomic DNA methylation in peripheral blood mononuclear cell DNA from 105 subjects homozygous for this mutation (T/T) and 187 homozygous for the wild-type (C/C) MTHFR genotype. The results show that genomic DNA methylation directly correlates with folate status and inversely with plasma homocysteine (tHcy) levels (P < 0.01). T/T genotypes had a diminished level of DNA methylation compared with those with the C/C wild-type (32.23 vs.62.24 ng 5 methylcytosine/microg DNA, P < 0.0001). When analyzed according to folate status, however, only the T/T subjects with low levels of folate accounted for the diminished DNA methylation (P < 0.0001). Moreover, in T/T subjects DNA methylation status correlated with the methylated proportion of red blood cell folate and was inversely related to the formylated proportion of red blood cell folates (P < 0.03) that is known to be solely represented in those individuals. These results indicate that the MTHFR C677T polymorphism influences DNA methylation status through an interaction with folate status. PMID- 11929968 TI - Self-assembly directed by NH...O hydrogen bonding: new layered molecular arrays derived from 4-tert- butylbenzoic acid and aliphatic diamines. AB - (1H) and (13)C NMR titrations in both CDCl(3) and CD(3)OD demonstrate that 4-tert butylbenzoic acid interacts with both propane-1,2-diamine and propane-1,3-diamine to yield 1:2 host-guest complexes in these solvents. Based on this observation, the isolation of new three-dimensional molecular arrays through cocrystallization of the above diamines and 4-tert-butylbenzoic acid (in a 1:2 molar ratio) has been achieved. X-ray studies of these self-assembled structures show that they incorporate [propane-1,2-diamine x (4-tert-butylbenzoic acid)(2)] or [propane-1,3 diamine x (4-tert-butylbenzoic acid)(2)] hydrogen-bonded motifs. Three structural derivatives of the latter type (two monohydrate forms and one anhydrous form) have been characterized. The structures are compared with a previously described three-dimensional array based on the "parent" [ethane-1,2-diamine x (benzoic acid)(2)] motif. Similarities occur between each of the structures. In each, a two-dimensional "ionic" layer consisting of an extensive network of hydrogen bonds is sandwiched between two "less polar" aromatic ring-containing layers. In the respective ionic layers, the carboxylic acid protons have been transferred onto the amines to yield diammonium cations, with all ammonium protons being involved in hydrogen bonding. In part, the adoption of these unusual layered structures seems to reflect a tendency toward maximization of both the number and strengths of the hydrogen bond interactions in the respective ionic layers. PMID- 11929969 TI - Unidirectional rotary motion in a liquid crystalline environment: color tuning by a molecular motor. AB - Life could not exist without motion induced by a variety of molecular motors. The construction of artificial motors by chemical synthesis, which can power motions that lead to macroscopic detectable effects in a system, is a major endeavor in contemporary science. To move toward this goal, a host-guest system, composed of a nematic liquid crystal film doped with a chiral light-driven molecular motor, is assembled. Irradiation of the film results in unidirectional rotary motion of the molecular motor, which induces a motion of the mesogenic molecules leading to a molecular reorganization and, as a consequence, a change in the color of the film. In this way, by control of the rotary motion at the molecular level, color tuning over the entire visible spectrum is achieved. These findings demonstrate that a molecular motor can exert a visually observable macroscopic change in a material. PMID- 11929970 TI - Toward complex matter: supramolecular chemistry and self-organization. PMID- 11929971 TI - Sialylation is essential for early development in mice. AB - Sialic acids are widely expressed as terminal carbohydrates on glycoconjugates of eukaryotic cells. Sialylation is crucial for a variety of cellular functions, such as cell adhesion or signal recognition, and regulates the biological stability of glycoproteins. The key enzyme of sialic acid biosynthesis is the bifunctional UDP-N-acetylglucosamine-2-epimerase/N-acetylmannosamine kinase (UDP GlcNAc 2-epimerase), which catalyzes the first two steps of sialic acid biosynthesis in the cytosol. We report that inactivation of the UDP-GlcNAc 2 epimerase by gene targeting causes early embryonic lethality in mice, thereby emphasizing the fundamental role of this bifunctional enzyme and sialylation during development. The need of UDP-GlcNAc 2-epimerase for a defined sialylation process is exemplified with the polysialylation of the neural cell adhesion molecule in embryonic stem cells. PMID- 11929972 TI - A protein subunit of human RNase P, Rpp14, and its interacting partner, OIP2, have 3'-->5' exoribonuclease activity. AB - The processing of precursor tRNAs at their 5' and 3' termini is a fundamental event in the biosynthesis of tRNA. RNase P is generally responsible for endonucleolytic removal of a leader sequence of precursor tRNA to generate the mature 5' terminus. However, much less is known about the RNase P counterparts or other proteins that are active at the tRNA 3' terminus. Here we show that one of the human RNase P subunits, Rpp14, together with one of its interacting protein partners, OIP2, is a 3'-->5' exoribonuclease with a phosphorolytic activity that processes the 3' terminus of precursor tRNA. Immunoprecipitates of a crude human RNase P complex can process both ends of precursor tRNA by hydrolysis, but purified RNase P has no exonuclease activity. Rpp14 and OIP2 may be part of an exosome activity. PMID- 11929973 TI - Molecular self-assembly of surfactant-like peptides to form nanotubes and nanovesicles. AB - Several surfactant-like peptides undergo self-assembly to form nanotubes and nanovesicles having an average diameter of 30-50 nm with a helical twist. The peptide monomer contains 7-8 residues and has a hydrophilic head composed of aspartic acid and a tail of hydrophobic amino acids such as alanine, valine, or leucine. The length of each peptide is approximately equal to 2 nm, similar to that of biological phospholipids. Dynamic light-scattering studies showed structures with very discrete sizes. The distribution becomes broader over time, indicating a very dynamic process of assembly and disassembly. Visualization with transmission electron microscopy of quick-freeze/deep-etch sample preparation revealed a network of open-ended nanotubes and some vesicles, with the latter being able to "fuse" and "bud" out of the former. The structures showed some tail sequence preference. Many three-way junctions that may act as links between the nanotubes have been observed also. Studies of peptide surfactant molecules have significant implications in the design of nonlipid biological surfactants and the understanding of the complexity and dynamics of the self-assembly processes. PMID- 11929974 TI - The Saccharomyces cerevisiae Fin1 protein forms cell cycle-specific filaments between spindle pole bodies. AB - The FIN1 gene from the yeast Saccharomyces cerevisiae encodes a basic protein with putative coiled-coil regions. Here we show that in large-budded cells a green fluorescent protein-Fin1 fusion protein is visible as a filament between the two spindle pole bodies. In resting cells the protein is undetectable, and in small-budded cells it is localized in the nucleus. During late mitosis it localizes on the spindle pole bodies. Filaments of cyano fluorescent protein tagged Fin1 colocalize with filaments of green fluorescent protein-tagged Tub1 only in large-budded cells. By electron and atomic force microscopy we showed that purified recombinant Fin1p self-assembles into filaments with a diameter of approximately 10 nm. Our results indicate that the Fin1 protein forms a cell cycle-specific filament, additional to the microtubules, between the spindle pole bodies of dividing yeast cells. PMID- 11929975 TI - Genetic analysis of the archaeon Methanosarcina barkeri Fusaro reveals a central role for Ech hydrogenase and ferredoxin in methanogenesis and carbon fixation. AB - Ech hydrogenase (Ech) from the methanogenic archaeon Methanosarcina barkeri catalyzes the reversible reduction of ferredoxin by H(2) and is a member of a distinct group of membrane-bound [NiFe] hydrogenases with sequence similarity to energy-conserving NADH:quinone oxidoreductase (complex I). To elucidate the physiological role(s) of Ech a mutant lacking this enzyme was constructed. The mutant was unable to grow on methanol/H(2)/CO(2), H(2)/CO(2), or acetate as carbon and energy sources but showed wild-type growth rates with methanol as sole substrate. Addition of pyruvate to the growth medium restored growth on methanol/H(2)/CO(2) but not on H(2)/CO(2) or acetate. Results obtained from growth experiments, cell suspension experiments, and enzyme activity measurements in cell extracts provide compelling evidence for essential functions of Ech and a 2[4Fe-4S] ferredoxin in the metabolism of M. barkeri. The following conclusions were made. (i) In acetoclastic methanogenesis, Ech catalyzes H(2) formation from reduced ferredoxin, generated by the oxidation of the carbonyl group of acetate to CO(2). (ii) Under autotrophic growth conditions, the enzyme catalyzes the energetically unfavorable reduction of ferredoxin by H(2), most probably driven by reversed electron transport, and the reduced ferredoxin thus generated functions as low potential electron donor for the synthesis of pyruvate in an anabolic pathway. (iii) Reduced ferredoxin in addition provides the reducing equivalents for the first step of methanogenesis from H(2)/CO(2), the reduction of CO(2) to formylmethanofuran. Thus, in vivo genetic analysis has led to the identification of the electron donor of this key initial step of methanogenesis. PMID- 11929976 TI - Configurationally homogeneous diastereomers of a linear hexa(tertiary phosphine): enantioselective self-assembly of a double-stranded parallel helicate of the type (P)-[Cu(3)(hexaphos)(2)](PF(6))(3). AB - Three configurationally homogeneous diastereomers of the linear hexa(tertiary phosphine) Ph(2)PCH(2)CH(2)P(Ph)CH(2)CH(2)P(Ph)CH(2)CH(2)P(Ph)CH(2)CH(2)P(Ph)CH(2)CH(2)PPh(2 (hexaphos) have been isolated in enantiomerically pure form, namely (R,S,S,R)-, (R,S,S,S)-, and (S,S,S,S)-hexaphos. The strongly helicating (R,S,S,R)-(-) form of the ligand combines with copper(I) ions to generate by stereoselective self assembly the P enantiomer of a parallel helicate of the type [Cu(3)(hexaphos)(2)](PF(6))(3), which has been characterized by x-ray crystallography. Theoretical modeling of the cation indicates that it is the relationship between the helicities of the two 10-membered rings containing the three copper ions, each of which has the twist-boat-chair-boat conformation, and the configurations of the three chiral, tetrahedral copper stereocenters of P configuration that determines the stereochemistry of the parallel and double alpha-helix conformers of the double-stranded trinuclear metal helicate. PMID- 11929977 TI - The plant-specific function of 2-Cys peroxiredoxin-mediated detoxification of peroxides in the redox-hierarchy of photosynthetic electron flux. AB - The 2-cysteine peroxiredoxins (2-Cys Prx) constitute an ancient family of peroxide detoxifying enzymes and have acquired a plant-specific function in the oxygenic environment of the chloroplast. Immunocytochemical analysis and work with isolated intact chloroplasts revealed a reversible binding of the oligomeric form of 2-Cys Prx to the thylakoid membrane. The oligomeric form of the enzyme was enhanced under stress. The 2-Cys Prx has a broad substrate specificity with activity toward hydrogen peroxides and complex alkyl hydroperoxides. During the peroxide reduction reaction, 2-Cys Prx is alternatively oxidized and reduced as it catalyzes an electron flow from an electron donor to peroxide. Escherichia coli thioredoxin, but also spinach thioredoxin f and m were able to reduce oxidized 2-Cys Prx. The midpoint redox potential of -315 mV places 2-Cys Prx reduction after Calvin cycle activation and before switching the malate valve for export of excess reduction equivalents to the cytosol. Thus the 2-Cys Prx has a defined and preferential place in the hierarchy of photosynthetic electron transport. The activity of 2-Cys Prx also is linked to chloroplastic NAD(P)H metabolism as indicated by the presence of the reduced form of the enzyme after feeding dihydroxyacetone phosphate to intact chloroplasts. The function of the 2 Cys Prx is therefore not confined to its role in the water-water cycle pathway for energy dissipation in photosynthesis but also mediates peroxide detoxification in the plastids during the dark phase. PMID- 11929978 TI - Hierarchical formation of helical supramolecular polymers via stacking of hydrogen-bonded pairs in water. AB - Bifunctional ureido-s-triazines provided with penta(ethylene oxide) side chains are able to self assemble in water, leading to helical columns via cooperative stacking of the hydrogen-bonded pairs (DADA array). Monofunctional ureido-s triazines do not form such helical architectures. The presence of a linker, covalently connecting the two ureido-s-triazine units, is essential as it generates a high local concentration of aromatic units, favorable for stacking interactions. This hydrophobic stacking of the aromatic units occurs at concentrations as low as 5 x 10(-6) M and can be visualized by using fluorescence spectroscopy. The stacking generates a hydrophobic microenvironment that allows intermolecular hydrogen bonding to occur at higher concentrations because the hydrogen bonds are shielded from competitive hydrogen bonding with water. This hierarchical process results in the formation of a helical self-assembled polymer in water at concentrations above 10(-4) M. Chiral side chains attached to the ureido-s-triazine units bias the helicity of these columns as concluded from CD spectroscopy and "Sergeants and Soldiers" experiments. PMID- 11929979 TI - Self-organizing super-structures formed from hydrogen-bonded biimidazolate metal complexes. AB - Manipulation of molecular crystals formed from self-organization is one of the important methods to develop the new molecular functional materials. In particular, we look upon mutual cohesive interactions of hydrogen bonding and metal coordination as one useful tool to construct the preprogramming superstructures. In this study, five controlled superstructures of a one dimensional linear chain--a zigzag ribbon, right-handed and left-handed helices, and a two-dimensional honeycomb sheet--are newly created by using the neutral metal complexes with some 2,2'-biimidazolate mono-anions. PMID- 11929980 TI - Unraveling the nanostructure of supramolecular assemblies of hydrogen-bonded rosettes on graphite: an atomic force microscopy study. AB - The self-organization of multicomponent tetrarosette assemblies into ordered nanostructures on graphite surfaces has been studied by atomic force microscopy (AFM). Real-space information on the level of individual molecules allowed us to analyze the underlying structure in unprecedented detail. In highly ordered nanorod domains, tetrarosettes 1(3) x (DEB)(12) arrange in the form of parallel rows with a spacing of 4.6 +/- 0.1 nm. High resolution AFM revealed the internal packing of the tetrarosette assemblies in these rows, which can be described by an oblique lattice with a = 2.5 +/- 0.3 nm, b = 5.0 +/- 0.1 nm, and gamma = 122 +/- 3 degrees. The results, together with recent improvements in synthetic approaches, contribute to the development of a general strategy to develop H bonding-based nanostructures with molecular precision. PMID- 11929981 TI - Peptide-amphiphile nanofibers: a versatile scaffold for the preparation of self assembling materials. AB - Twelve derivatives of peptide-amphiphile molecules, designed to self-assemble into nanofibers, are described. The scope of amino acid selection and alkyl tail modification in the peptide-amphiphile molecules are investigated, yielding nanofibers varying in morphology, surface chemistry, and potential bioactivity. The results demonstrate the chemically versatile nature of this supramolecular system and its high potential for manufacturing nanomaterials. In addition, three different modes of self-assembly resulting in nanofibers are described, including pH control, divalent ion induction, and concentration. PMID- 11929982 TI - Presence of mouse mammary tumor virus specifically alters the body odor of mice. AB - It has long been recognized that various genetic and metabolic human disorders alter body odor, which is not surprising because they may alter body chemistry. Thus, it has been suggested that some human diseases may be diagnosed by odor alone. In that regard, the mouse mammary tumor virus (MMTV) and its tumors of mice, which may have human counterparts, are of special interest because of the need for basic research possible only in inbred and genetically defined animals. Accordingly, we now show that the mouse MMTV, whether obtained environmentally or genetically transmitted, alters the body odor of mice in both males and females, and regardless of the presence or absence of tumors. These observations, together with the prospect of artificial human odor discrimination, may aid in the search for early human diagnostics. PMID- 11929983 TI - Catalysis of cis/trans isomerization in native HIV-1 capsid by human cyclophilin A. AB - Packaging of cyclophilin A (CypA) into HIV-1 virions is essential for efficient replication; however, the reason for this is unknown. Incorporation is mediated through binding to the Gly-89-Pro-90 peptide bond of the N-terminal domain of HIV 1 capsid (CA(N)). Despite the fact that CypA is a peptidyl-prolyl cis/trans isomerase, catalytic activity on CA(N) has not been observed previously. We show here, using NMR exchange spectroscopy, that CypA does not only bind to CA(N) but also catalyzes efficiently the cis/trans isomerization of the Gly-89-Pro-90 peptide bond. In addition, conformational changes in CA(N) distal to the CypA binding loop are observed on CypA binding and catalysis. The results provide experimental evidence for efficient CypA catalysis on a natively folded and biologically relevant protein substrate. PMID- 11929984 TI - CD8(+) T cells are not necessary for 1 alpha,25-dihydroxyvitamin D(3) to suppress experimental autoimmune encephalomyelitis in mice. AB - In addition to its role in calcium and phosphorous homeostasis, 1 alpha,25 dihydroxyvitamin D(3) [1,25-(OH)(2)D(3)] appears to be a modulator of the immune system. Administration of 1,25-(OH)(2)D(3) prevents disease in several autoimmune animal models, including experimental autoimmune encephalomyelitis (EAE). The vitamin D receptor is believed to mediate this activity. Among cells of the immune system, CD8(+) T cells have the highest levels of the vitamin D receptor. Because CD8(+) T cells have been implicated as both suppressors and effectors of the inflammation associated with multiple sclerosis and EAE, we examined the question of whether the 1,25-(OH)(2)D(3) suppression of EAE occurs through a CD8(+) T cell-dependent mechanism. To test this hypothesis, mice that are homozygous knockouts for the alpha chain of the CD8 receptor and have been characterized as lacking functional CD8(+) T cells (CD8(+) -/-) were provided 1,25-(OH)(2)D(3) in their diet before EAE induction. Although CD8(+) -/- mice fed the same diet lacking 1,25-(OH)(2)D(3) have a high incidence of EAE, EAE did not occur in CD8(+) -/- mice fed the diet containing 1,25-(OH)(2)D(3). We conclude that CD8(+) T cells neither are needed nor do they play a role in the prevention of EAE by 1,25-(OH)(2)D(3). PMID- 11929985 TI - CD40 stimulation leads to effective therapy of CD40(-) tumors through induction of strong systemic cytotoxic T lymphocyte immunity. AB - Adequate spontaneous activation of tumor-specific T lymphocytes in tumor-bearing hosts is rare, despite the expression of tumor antigens that are potentially highly immunogenic. For example, failure of the immune system to raise competent responses against established tumors expressing the human adenovirus E1A-antigen allows this tumor to grow in immunocompetent mice. We show that systemic in vivo administration of agonistic anti-CD40 antibodies into tumor-bearing mice results in tumor eradication mediated by CD8(+) T cells. Treatment resulted in a strong expansion and systemic accumulation of E1A-specific CTL and depended on CD40 expression on host cells, as the tumor was CD40(-), and therapy failed in CD40 deficient mice. Local intratumoral administration of anti-CD40 mAb is equally effective in licensing strong, systemic CTL immunity, resulting in the clearance of distant tumor nodules. Our data indicate that the immune response after cancer host interactions can be directed toward competence, leading to the cure of established tumors merely by delivery of a CD40-dependent "license to kill" signal. PMID- 11929986 TI - Inhibition of chymotrypsin through surface binding using nanoparticle-based receptors. AB - Efficient binding of biomacromolecular surfaces by synthetic systems requires the effective presentation of complementary elements over large surface areas. We demonstrate here the use of mixed monolayer protected gold clusters (MMPCs) as scaffolds for the binding and inhibition of chymotrypsin. In these studies anionically functionalized amphiphilic MMPCs were shown to inhibit chymotrypsin through a two-stage mechanism featuring fast reversible inhibition followed by a slower irreversible process. This interaction is very efficient, with a K(i)(app) = 10.4 +/- 1.3 nM. The MMPC-protein complex was characterized by CD, demonstrating an almost complete denaturation of the enzyme over time. Dynamic light scattering studies confirm that inhibition proceeds without substantial MMPC aggregation. The electrostatic nature of the engineered interactions provides a level of selectivity: little or no inhibition of function was observed with elastase, beta-galactosidase, or cellular retinoic acid binding protein. PMID- 11929987 TI - Imaging the progression of Alzheimer pathology through the brain. PMID- 11929988 TI - From bacteria to mitochondria: aconitase yields surprises. PMID- 11929989 TI - What are big brains for? PMID- 11929990 TI - Dynamics of self assembly of magnetized disks rotating at the liquid-air interface. AB - This paper is a theoretical study of dynamic self assembly in a system of millimeter-sized magnetized disks floating at a liquid-air interface and spinning under the influence of a rotating magnetic field. Equations of motions are derived that account for the hydrodynamic and magnetic forces acting in the system. Numerical integration of these equations predicts formation of ordered structures of spinning disks; the simulated structures reproduce the patterns observed experimentally. PMID- 11929992 TI - The Pythagorean Theorem: I. The finite case. AB - The Pythagorean Theorem and variants of it are studied. The variations evolve to a formulation in terms of noncommutative, conditional expectations on von Neumann algebras that displays the theorem as the basic result of noncommutative, metric, Euclidean Geometry. The emphasis in the present article is finite dimensionality, both "discrete" and "continuous." PMID- 11929991 TI - Phosphate backbone neutralization increases duplex DNA flexibility: a model for protein binding. AB - An important component of protein-DNA recognition is the charge neutralization of DNA backbone phosphates and subsequent protein-induced DNA bending. Replacement of phosphates by neutral methylphosphonates has previously been shown to be a model for protein-induced bending. In addition to bending, the neutralization process may change the inherent flexibility of the DNA--a feature never before tested. We have developed a method to measure the differential flexibility of duplex DNA when methylphosphonate substitutions are made and find that the local flexibility is increased up to 40%. These results imply that backbone neutralization-dependent DNA flexibility augments DNA-binding motifs in protein DNA recognition processes. PMID- 11929993 TI - The in vivo function of the ribosome-associated Hsp70, Ssz1, does not require its putative peptide-binding domain. AB - Two proteins of the Hsp70 class (Ssb and Ssz1) and one of the J-type class (Zuo1) of molecular chaperones reside on the yeast ribosome, with Ssz1 forming a stable heterodimer with Zuo1. We designed experiments to address the roles of these two distantly related ribosome-associated Hsp70s and their functional relationship to Zuo1. Strains lacking all three proteins have the same phenotype as those lacking only one, suggesting that these chaperones all function in the same pathway. The Hsp70 Ssb, whose peptide-binding domain is essential for its in vivo function, can be crosslinked to nascent chains on ribosomes that are as short as 54 amino acids, suggesting that Ssb interacts with nascent chains that extend only a short distance beyond the tunnel of the ribosome. A ssz1 mutant protein lacking its putative peptide-binding domain allows normal growth. Thus, binding of unfolded protein substrates in a manner similar to that of typical Hsp70s is not critical for Ssz1's in vivo function. The three chaperones are present in cells in approximately equimolar amounts compared with ribosomes. The level of Ssb can be reduced only a few-fold before growth is affected. However, a 50- to 100-fold reduction of Ssz1 and Zuo1 levels does not have a substantial effect on cell growth. On the basis of these results, we propose that Ssbs function as the major Hsp70 chaperone for nascent chains on the ribosome, and that Ssz1 has evolved to perform a nonclassical function, perhaps modulating Zuo1's ability to function as a J-type chaperone partner of Ssb. PMID- 11929994 TI - A functional chaperone triad on the yeast ribosome. AB - The chaperones RAC (ribosome-associated complex), consisting of Ssz1p and zuotin, and Ssb1/2p are associated with ribosomes of yeast. Ssb1/2p was previously shown to form a crosslink product to polypeptides trapped in ribosome-nascent chain complexes (RNCs) in vitro. Here we show that an efficient crosslink of the nascent chain to Ssb1/2p depends on the presence of functional RAC. The crosslink to Ssb1/2p was significantly diminished if (i) RAC was removed from RNCs: a process reversed by addition of purified RAC; (ii) RAC carried a mutation in the J-domain of zuotin, leading to its inactivation in vivo; (iii) RAC's Ssz1p subunit was absent because RNCs were generated in a Deltassz1-derived translation extract. In vivo the same specific set of growth defects caused by the absence of any of the three chaperones was also displayed by a Deltassb1/2Deltassz1Deltazuo1 strain. The combination of in vitro and in vivo data supports a model in which Ssb1/2p, Ssz1p, and zuotin act in concert on nascent chains while they are being synthesized. PMID- 11929996 TI - Diversity and evolution of the green fluorescent protein family. AB - The family of proteins homologous to the green fluorescent protein (GFP) from Aequorea victoria exhibits striking diversity of features, including several different types of autocatalytically synthesized chromophores. Here we report 11 new members of the family, among which there are 3 red-emitters possessing unusual features, and discuss the similarity relationships within the family in structural, spectroscopic, and evolutionary terms. Phylogenetic analysis has shown that GFP-like proteins from representatives of subclass Zoantharia fall into at least four distinct clades, each clade containing proteins of more than one emission color. This topology suggests multiple recent events of color conversion. Combining this result with previous mutagenesis and structural data, we propose that (i) different chromophore structures are alternative products synthesized within a similar autocatalytic environment, and (ii) the phylogenetic pattern and color diversity in reef Anthozoa is a result of a balance between selection for GFP-like proteins of particular colors and mutation pressure driving the color conversions. PMID- 11929995 TI - Selenoprotein R is a zinc-containing stereo-specific methionine sulfoxide reductase. AB - Selenoprotein R (SelR) is a mammalian selenocysteine-containing protein with no known function. Here we report that cysteine homologs of SelR are present in all organisms except certain parasites and hyperthermophiles, and this pattern of occurrence closely matches that of only one protein, peptide methionine sulfoxide reductase (MsrA). Moreover, in several genomes, SelR and MsrA genes are fused or clustered, and their expression patterns suggest a role of both proteins in protection against oxidative stress. Consistent with these computational screens, growth of Saccharomyces cerevisiae SelR and MsrA mutant strains was inhibited, and the strain lacking both genes could not grow, in the presence of H2O2 and methionine sulfoxide. We found that the cysteine mutant of mouse SelR, as well as the Drosophila SelR homolog, contained zinc and reduced methionine-R-sulfoxide, but not methionine-S-sulfoxide, in in vitro assays, a function that is both distinct and complementary to the stereo-specific activity of MsrA. These findings identify a function of the conserved SelR enzyme family, define a pathway of methionine sulfoxide reduction, reveal a case of convergent evolution of similar function in structurally distinct enzymes, and suggest a previously uncharacterized redox regulatory role of selenium in mammals. PMID- 11929997 TI - Rapid compaction during RNA folding. AB - We have used small angle x-ray scattering and computer simulations with a coarse grained model to provide a time-resolved picture of the global folding process of the Tetrahymena group I RNA over a time window of more than five orders of magnitude. A substantial phase of compaction is observed on the low millisecond timescale, and the overall compaction and global shape changes are largely complete within one second, earlier than any known tertiary contacts are formed. This finding indicates that the RNA forms a nonspecifically collapsed intermediate and then searches for its tertiary contacts within a highly restricted subset of conformational space. The collapsed intermediate early in folding of this RNA is grossly akin to molten globule intermediates in protein folding. PMID- 11929998 TI - Fluorescence polarization of green fluorescence protein. AB - We report here the striking anisotropy of fluorescence exhibited by crystals of native green fluorescence protein (GFP). The crystals were generated by water dialysis of highly purified GFP obtained from the jellyfish Aequorea. We find that the fluorescence becomes six times brighter when the excitation, or emission, beam is polarized parallel (compared with perpendicular) to the crystal long axis. Thus, the major dipoles of the fluorophores must be oriented very nearly parallel to the crystal long axis. Observed in a polarizing microscope between parallel polars instead of either a polarizer or analyzer alone, the fluorescence polarization ratio rises to an unexpectedly high value of about 30:1, nearly the product of the fluorescence excitation and emission ratios, suggesting a sensitive method for measuring fluorophore orientations, even of a single fluorophore molecule. We have derived equations that accurately describe the relative fluorescence intensities of crystals oriented in various directions, with the polarizer and analyzer arranged in different configurations. The equations yield relative absorption and fluorescence coefficients for the four transition dipoles involved. Finally, we propose a model in which the elongated crystal is made of GFP molecules that are tilted 60 degrees to align the fluorophores parallel to the crystal long axis. The unit layer in the model may well correspond to the arrangement of functional GFP molecules, to which resonant energy is efficiently transmitted from Ca2+-activated aequorin, in the jellyfish photophores. PMID- 11929999 TI - Mg2+-dependent conformational change of RNA studied by fluorescence correlation and FRET on immobilized single molecules. AB - Fluorescence correlation spectroscopy (FCS) of fluorescence resonant energy transfer (FRET) on immobilized individual fluorophores was used to study the Mg2+ facilitated conformational change of an RNA three-helix junction, a structural element that initiates the folding of the 30S ribosomal subunit. Transitions of the RNA junction between open and folded conformations resulted in fluctuations in fluorescence by FRET. Fluorescence fluctuations occurring between two FRET states on the millisecond time scale were found to be dependent on Mg2+ and Na+ concentrations. Correlation functions of the fluctuations were used to determine transition rates between the two conformations as a function of Mg2+ or Na+ concentration. Both the opening and folding rates were found to vary with changing salt conditions. Assuming specific binding of divalent ions to RNA, the Mg2+ dependence of the observed rates cannot be explained by conformational change induced by Mg2+ binding/unbinding, but is consistent with a model in which the intrinsic conformational change of the RNA junction is altered by uptake of Mg2+ ion(s). This version of FCS/FRET on immobilized single molecules is demonstrated to be a powerful technique in the study of conformational dynamics of biomolecules over time scales ranging from microseconds to seconds. PMID- 11930000 TI - FKBP12-rapamycin-associated protein associates with mitochondria and senses osmotic stress via mitochondrial dysfunction. AB - FKBP12-rapamycin associated protein (FRAP, also known as mTOR or RAFT) is the founding member of the phosphatidylinositol kinase-related kinase family and functions as a sensor of physiological signals that regulate cell growth. Signals integrated by FRAP include nutrients, cAMP levels, and osmotic stress, and cellular processes affected by FRAP include transcription, translation, and autophagy. The mechanisms underlying the integration of such diverse signals by FRAP are largely unknown. Recently, FRAP has been reported to be regulated by mitochondrial dysfunction and depletion of ATP levels. Here we show that exposure of cells to hyperosmotic conditions (and to glucose-deficient growth medium) results in rapid and reversible dissipation of the mitochondrial proton gradient. These results suggest that the ability of FRAP to mediate osmotic stress response (and glucose deprivation response) is by means of an intermediate mitochondrial dysfunction. We also show that in addition to cytosolic FRAP a large portion of FRAP associates with the mitochondrial outer membrane. The results support the existence of a stress-sensing module consisting of mitochondria and mitochondrial outer membrane-associated FRAP. This module allows the cell to integrate a variety of stress signals that affect mitochondrial function and regulate a growth checkpoint involving p70 S6 kinase. PMID- 11930001 TI - The cAMP-specific phosphodiesterase TbPDE2C is an essential enzyme in bloodstream form Trypanosoma brucei. AB - Chemotherapy of human sleeping sickness, a fatal disease caused by the protozoan parasite Trypanosoma brucei, is in a dismal state, and the identification and characterization of new drug targets is an urgent prerequisite for an improvement of the dramatic situation in the field. Over the last several years, inhibitors of cyclic nucleotide-specific phosphodiesterases have proven to be highly successful drug candidates for an assortment of clinical conditions. Their potential as antiparasitic drugs has not been explored so far. This study reports the characterization of a cAMP-specific phosphodiesterase from T. brucei, TbPDE2C. This enzyme is a class I phosphodiesterase, and it is a member of a small enzyme family in T. brucei, TbPDE2. Inhibitors of this enzyme block the proliferation of bloodstream form trypanosomes in culture. RNA interference experiments demonstrated that the TbPDE2 family, and in particular TbPDE2C, are essential for maintaining intracellular cAMP concentrations within a physiological range. Bloodstream form trypanosomes are exquisitely sensitive to elevated concentrations of intracellular cAMP, and a disruption of TbPDE2C function quickly leads to the disruption of nuclear and cellular cell division, and to cell death. TbPDE2C might represent a novel drug target for the development of new and effective trypanocidal drugs. PMID- 11930002 TI - Renewable and nonrenewable resources: amino acid turnover and allocation to reproduction in Lepidoptera. AB - The allocation of nutritional resources to reproduction in animals is a complex process of great evolutionary significance. We use compound-specific stable isotope analysis of carbon (GC/combustion/isotope ratio MS) to investigate the dietary sources of egg amino acids in a nectar-feeding hawkmoth. Previous work suggests that the nutrients used in egg manufacture fall into two classes: those that are increasingly synthesized from adult dietary sugar over a female's lifetime (renewable resources), and those that remain exclusively larval in origin (nonrenewable resources). We predict that nonessential and essential amino acids correspond to these nutrient classes and test this prediction by analyzing egg amino acids from females fed isotopically distinct diets as larvae and as adults. The results demonstrate that essential egg amino acids originate entirely from the larval diet. In contrast, nonessential egg amino acids were increasingly synthesized from adult dietary sugars, following a turnover pattern across a female's lifetime. This study demonstrates that female Lepidoptera can synthesize a large fraction of egg amino acids from nectar sugars, using endogenous sources of nitrogen. However, essential amino acids derive only from the larval diet, placing an upper limit on the use of adult dietary resources to enhance reproductive success. PMID- 11930003 TI - Evolutionary conservation of chromosome territory arrangements in cell nuclei from higher primates. AB - We demonstrate that the nuclear topological arrangement of chromosome territories (CTs) has been conserved during primate evolution over a period of about 30 million years. Recent evidence shows that the positioning of chromatin in human lymphocyte nuclei is correlated with gene density. For example, human chromosome 19 territories, which contain mainly gene-dense and early replicating chromatin, are located toward the nuclear center, whereas chromosome 18 territories, which consist mainly of gene-poor and later replicating chromatin, is located close to the nuclear border. In this study, we subjected seven different primate species to comparative analysis of the radial distribution pattern of human chromosome 18 and 19-homologous chromatin by three-dimensional fluorescence in situ hybridization. Our data demonstrate that gene-density-correlated radial chromatin arrangements were conserved during higher-primate genome evolution, irrespective of the major karyotypic rearrangements that occurred in different phylogenetic lineages. The evolutionarily conserved positioning of homologous chromosomes or chromosome segments in related species supports evidence for a functionally relevant higher-order chromatin arrangement that is correlated with gene-density. PMID- 11930004 TI - Sexual selection driving diversification in jumping spiders. AB - Theory predicts that speciation rates should be accelerated in organisms undergoing sexual selection. In systems involving female choice, sexual selection acts directly on traits that may be important in prezygotic reproductive isolation, potentially fostering rapid divergence of such traits among allopatric populations. Despite the appeal of this concept, it has proven difficult to document. We provide genetic, behavioral, and simulation data illustrating that the striking and possibly recent divergence in traits of male behavior and morphology among populations of the jumping spider Habronattus pugillis can be attributed to sexual selection. We have found evidence for varying degrees of lower female response and offspring viability among some between-population crosses, consistent with the early stages of speciation. We have developed a gene tree-based method for comparing phenotypic and genetic data sets to infer selection, and have found robust statistical evidence that directional selection has acted on male traits, by confirming that their rate of fixation exceeds that of neutral mitochondrial genes. Because these traits are apparent targets of female choice, the results indicate that sexual selection is driving divergence of phenotypes potentially crucial to the speciation process. PMID- 11930005 TI - Mitf and Tfe3, two members of the Mitf-Tfe family of bHLH-Zip transcription factors, have important but functionally redundant roles in osteoclast development. AB - The Mitf-Tfe family of basic helix-loop-helix-leucine zipper (bHLH-Zip) transcription factors encodes four family members: Mitf, Tfe3, Tfeb, and Tfec. In vitro, each protein in the family can bind DNA as a homo- or heterodimer with other family members. Mutational studies in mice have shown that Mitf is essential for melanocyte and eye development, whereas Tfeb is required for placental vascularization. Here, we uncover a role for Tfe3 in osteoclast development, a role that is functionally redundant with Mitf. Although osteoclasts seem normal in Mitf or Tfe3 null mice, the combined loss of the two genes results in severe osteopetrosis. We also show that Tfec mutant mice are phenotypically normal, and that the Tfec mutation does not alter the phenotype of Mitf, Tfeb, or Tfe3 mutant mice. Surprisingly, our studies failed to identify any phenotypic overlap between the different Mitf-Tfe mutations. These results suggest that heterodimeric interactions are not essential for Mitf-Tfe function in contrast to other bHLH-Zip families like Myc/Max/Mad, where heterodimeric interactions seem to be essential. PMID- 11930006 TI - The combined absence of NF-kappa B1 and c-Rel reveals that overlapping roles for these transcription factors in the B cell lineage are restricted to the activation and function of mature cells. AB - Transcription factors NF-kappaB1 and c-Rel, individually dispensable during embryogenesis, serve similar, yet distinct, roles in the function of mature hemopoietic cells. Redundancy among Rel/NF-kappaB family members prompted an examination of the combined roles of c-Rel and NF-kappaB1 by using mice that lack both proteins. Embryonic development and the maturation of hemopoietic progenitors were unaffected in nfkb1(-/-)c-rel(-/-) mice. Peripheral T cell populations developed normally, but follicular, marginal zone, and CD5(+) peritoneal B cell populations all were reduced. In culture, a failure of mitogen stimulated nfkb1(-/-)c-rel(-/-) B cells to proliferate was caused by a cell cycle defect in early G(1) that prevented growth. In vivo, defects in humoral immunity and splenic architecture seen in nfkb1(-/-) and c-rel(-/-) mice were exacerbated in the double mutant mice. These findings demonstrate that in the B lineage overlapping roles for NF-kappaB1 and c-Rel appear to be restricted to regulating the activation and function of mature cells. PMID- 11930007 TI - Role of poly(ADP-ribosyl)ation in DNA-PKcs- independent V(D)J recombination. AB - V(D)J recombination is critical to the generation of a functional immune system. Intrinsic to the assembly of antigen receptor genes is the formation of endogenous DNA double-strand breaks, which normally are excluded from the cellular surveillance machinery because of their sequestration in a synaptic complex and/or rapid resolution. In cells deficient in double-strand break repair, such recombination-induced breaks fail to be joined promptly and therefore are at risk of being recognized as DNA damage. Poly(ADP-ribose) polymerase-1 is an important factor in the maintenance of genomic integrity and is believed to play a central role in DNA repair. Here we provide visual evidence that in a recombination inducible severe combined immunodeficient cell line poly(ADP-ribose) formation occurs during the resolution stage of V(D)J recombination where nascent opened coding ends are generated. Poly(ADP-ribose) formation appears to facilitate coding end resolution. Furthermore, formation of Mre11 foci coincide with these areas of poly(ADP-ribosyl)ation. In contrast, such a response is not observed in wild-type cells possessing a functional catalytic subunit of DNA-dependent protein kinase (DNA-PK(cs)). Thus, V(D)J recombination invokes a DNA damage response in cells lacking DNA-PK(cs) activity, which in turn promotes DNA-PK(cs)-independent resolution of recombination intermediates. PMID- 11930008 TI - Osteopontin deficiency protects joints against destruction in anti-type II collagen antibody-induced arthritis in mice. AB - Rheumatoid arthritis is one of the most critical diseases that impair the quality of life of patients, but its pathogenesis has not yet been fully understood. Osteopontin (OPN) is an extracellular matrix protein containing Arg-Gly-Asp (RGD) sequence, which interacts with alpha(v)beta3 integrins, promotes cell attachment, and cell migration and is expressed in both synovial cells and chondrocytes in rheumatoid arthritis; however, its functional relationship to arthritis has not been known. Therefore, we investigated the roles of OPN in the pathogenesis of inflammatory process in a rheumatoid arthritis model induced by a mixture of anti type II collagen mAbs and lipopolysaccharide (mAbs/LPS). mAbs/LPS injection induced OPN expression in synovia as well as cartilage, and this expression was associated with joint swelling, destruction of the surface structures of the joint based on scanning electron microscopy, and loss of toluidine blue-positive proteoglycan content in the articular cartilage in wild-type mice. In contrast, OPN deficiency prevented the mice from such surface destruction, loss of proteoglycan in the articular joint cartilage, and swelling of the joints even when the mice were subjected to mAbs/LPS injection. Furthermore, mAbs/LPS injection in wild-type mice enhanced the levels of CD31-positive vessels in synovia and terminal deoxynucleotidyltransferase-mediated UTP end labeling positive chondrocytes in the articular cartilage, whereas such angiogenesis as well as chondrocyte apoptosis was suppressed significantly in OPN-deficient mice. These results indicated that OPN plays a critical role in the destruction of joint cartilage in the rheumatoid arthritis model in mice via promotion of angiogenesis and induction of chondrocyte apoptosis. PMID- 11930009 TI - Panhandle and reverse-panhandle PCR enable cloning of der(11) and der(other) genomic breakpoint junctions of MLL translocations and identify complex translocation of MLL, AF-4, and CDK6. AB - We used panhandle PCR to clone the der(11) genomic breakpoint junction in three leukemias with t(4;11) and devised reverse-panhandle PCR to clone the breakpoint junction of the other derivative chromosome. This work contributes two elements to knowledge on MLL translocations. First is reverse-panhandle PCR for cloning breakpoint junctions of the other derivative chromosomes, sequences of which are germane to understanding the MLL translocation process. The technique revealed duplicated sequences in one case of infant acute lymphoblastic leukemia (ALL) and small deletions in a case of treatment-related ALL. The second element is discovery of a three-way rearrangement of MLL, AF-4, and CDK6 in another case of infant ALL. Cytogenetic analysis was unsuccessful at diagnosis, but suggested t(4;11) and del(7)(q21q31) at relapse. Panhandle PCR analysis of the diagnostic marrow identified a breakpoint junction of MLL intron 8 and AF-4 intron 3. Reverse-panhandle PCR identified a breakpoint junction of CDK6 from band 7q21-q22 and MLL intron 9. CDK6 encodes a critical cell cycle regulator and is the first gene of this type disrupted by MLL translocation. Cdk6 is overexpressed or disrupted by translocation in many cancers. The in-frame CDK6-MLL transcript is provocative with respect to a potential contribution of the predicted Cdk6-MLL fusion protein in the genesis of the ALL, which also contains an in-frame MLL-AF4 transcript. The sequences in these three cases show additional MLL genomic breakpoint heterogeneity. Each breakpoint junction suggests nonhomologous end joining and is consistent with DNA damage and repair. CDK6-MLL is a new fusion of both genes. PMID- 11930010 TI - Severe iron deficiency anemia in transgenic mice expressing liver hepcidin. AB - We recently reported the hemochromatosis-like phenotype observed in our Usf2 knockout mice. In these mice, as in murine models of hemochromatosis and patients with hereditary hemochromatosis, iron accumulates in parenchymal cells (in particular, liver and pancreas), whereas the reticuloendothelial system is spared from this iron loading. We suggested that this phenotypic trait could be attributed to the absence, in the Usf2 knockout mice, of a secreted liver specific peptide, hepcidin. We conjectured that the reverse situation, namely overexpression of hepcidin, might result in phenotypic traits of iron deficiency. This question was addressed by generating transgenic mice expressing hepcidin under the control of the liver-specific transthyretin promoter. We found that the majority of the transgenic mice were born with a pale skin and died within a few hours after birth. These transgenic animals had decreased body iron levels and presented severe microcytic hypochromic anemia. So far, three mosaic transgenic animals have survived. They were unequivocally identified by physical features, including reduced body size, pallor, hairless and crumpled skin. These pleiotropic effects were found to be associated with erythrocyte abnormalities, with marked anisocytosis, poikylocytosis and hypochromia, which are features characteristic of iron-deficiency anemia. These results strongly support the proposed role of hepcidin as a putative iron-regulatory hormone. The animal models devoid of hepcidin (the Usf2 knockout mice) or overexpressing the peptide (the transgenic mice presented in this paper) represent valuable tools for investigating iron homeostasis in vivo and for deciphering the molecular mechanisms of hepcidin action. PMID- 11930011 TI - Bradykinin antagonist dimer, CU201, inhibits the growth of human lung cancer cell lines by a "biased agonist" mechanism. AB - All small cell (SCLCs) and many non-small cell lung cancers (NSCLCs) have neuroendocrine features including production of neuropeptides and cell surface receptors creating autocrine and paracrine growth loops. Neuropeptides bind to a family of 7-transmembrane receptors and activate heterotrimeric G proteins consisting of G(alphaq) and G(alpha12,13). Substance P derivatives (SPDs) induced apoptosis and inhibited growth of lung cancer cells by discoordinately inhibiting G(alphaq) and stimulating G(alpha12,13). However, these SPDs had low potency and short half-lives. In this report we show that a bradykinin antagonist dimer, CU201, inhibited the growth of SCLC and NSCLC cell lines with or without multidrug-resistant proteins and was 10-fold more potent with a longer plasma half-life than SPDs. Bradykinin agonists in either monomeric or dimeric form and monomeric bradykinin antagonist have no effect on lung cancer cell growth. The dimeric linking moiety of the two molecules was created, requiring a sufficient number of carbon chains to provide critical spacing between the two antagonists. CU201 inhibited intracellular Ca2+ release in response to bradykinin, indicating blockage of the G(alphaq) signal, and stimulated c-Jun kinases, indicating stimulation of the G(alpha12,13) pathway. CU201-induced apoptosis was preceded by unique changes in apparent nuclear DNA binding and by c-Jun kinase and caspase-3 activation. At the concentration at which CU201 inhibited the growth of the cancer cells, it had no effect on the growth of normal lung cells in vitro. CU201 and similar compounds offer hope of becoming a new form of targeted therapy for tumors with neuroendocrine properties. PMID- 11930012 TI - Genes directly controlled by CtrA, a master regulator of the Caulobacter cell cycle. AB - Studies of the genetic network that controls the Caulobacter cell cycle have identified a response regulator, CtrA, that controls, directly or indirectly, one quarter of the 553 cell cycle-regulated genes. We have performed in vivo genomic binding site analysis of the CtrA protein to identify which of these genes have regulatory regions bound directly by CtrA. By combining these data with previous global analysis of cell cycle transcription patterns and gene expression profiles of mutant ctrA strains, we have determined that CtrA directly regulates at least 95 genes. The total group of CtrA-regulated genes includes those involved in polar morphogenesis, DNA replication initiation, DNA methylation, cell division, and cell wall metabolism. Also among the genes in this notably large regulon are 14 that encode regulatory proteins, including 10 two-component signal transduction regulatory proteins. Identification of additional regulatory genes activated by CtrA will serve to directly connect new regulatory modules to the network controlling cell cycle progression. PMID- 11930013 TI - Genetic control of quorum-sensing signal turnover in Agrobacterium tumefaciens. AB - A signal turnover system is an essential component of many genetic regulatory mechanisms. The best-known example is the ubiquitin-dependent protein degradation system that exists in many organisms. We found that Agrobacterium tumefaciens adopts a unique signal turnover system to control exiting from a quorum-sensing mode. A. tumefaciens regulates Ti plasmid conjugal transfer by a quorum-sensing signal, N-3-oxo-octanoyl homoserine lactone (3OC8HSL), also known as Agrobacterium autoinducer. By using Tn5 mutagenesis and a functional cloning approach, we identified two genes that are involved in switching from a conjugal quorum-sensing mode to a nonconjugal mode at the onset of stationary phase. First, we located attJ, which codes for an IclR-type suppressor that regulates the second gene attM. The latter encodes a homologue of N-acylhomoserine lactone (AHL)-lactonase. Mass spectrometry analysis shows that the enzyme encoded by attM is an AHL-lactonase that hydrolyzes the lactone ring of 3OC8HSL. In wild-type A. tumefaciens, attM expression is initially suppressed by AttJ but significantly elevated at the stationary phase accompanied a sharp decline in 3OC8HSL. DNA gel retardation analysis shows that AttJ specifically binds to the promoter that controls AHL-lactonase expression. Mutation of attJ resulted in constitutive production of AHL-lactonase that abolishes 3OC8HSL accumulation and Ti plasmid transfer. These data suggest that A. tumefaciens has a sophisticated multicomponent quorum-sensing signal turnover system, allowing the cell to sense a change in growth and adjust cellular activities accordingly. PMID- 11930014 TI - The complete genome of hyperthermophile Methanopyrus kandleri AV19 and monophyly of archaeal methanogens. AB - We have determined the complete 1,694,969-nt sequence of the GC-rich genome of Methanopyrus kandleri by using a whole direct genome sequencing approach. This approach is based on unlinking of genomic DNA with the ThermoFidelase version of M. kandleri topoisomerase V and cycle sequencing directed by 2'-modified oligonucleotides (Fimers). Sequencing redundancy (3.3x) was sufficient to assemble the genome with less than one error per 40 kb. Using a combination of sequence database searches and coding potential prediction, 1,692 protein-coding genes and 39 genes for structural RNAs were identified. M. kandleri proteins show an unusually high content of negatively charged amino acids, which might be an adaptation to the high intracellular salinity. Previous phylogenetic analysis of 16S RNA suggested that M. kandleri belonged to a very deep branch, close to the root of the archaeal tree. However, genome comparisons indicate that, in both trees constructed using concatenated alignments of ribosomal proteins and trees based on gene content, M. kandleri consistently groups with other archaeal methanogens. M. kandleri shares the set of genes implicated in methanogenesis and, in part, its operon organization with Methanococcus jannaschii and Methanothermobacter thermoautotrophicum. These findings indicate that archaeal methanogens are monophyletic. A distinctive feature of M. kandleri is the paucity of proteins involved in signaling and regulation of gene expression. Also, M. kandleri appears to have fewer genes acquired via lateral transfer than other archaea. These features might reflect the extreme habitat of this organism. PMID- 11930015 TI - Gene expression analysis in schizophrenia: reproducible up-regulation of several members of the apolipoprotein L family located in a high-susceptibility locus for schizophrenia on chromosome 22. AB - We screened a custom-made candidate gene cDNA array comprising 300 genes. Genes chosen have either been implicated in schizophrenia, make conceptual sense in the light of the current understanding of the disease, or are located on high susceptibility chromosome locations. The array screen using prefrontal cortex tissue from 10 schizophrenia and 10 control brains revealed robust up-regulation of apolipoprotein L1 (apo L1) by 2.6-fold. The finding was cross-validated in a blinded quantitative PCR study using prefrontal cortex tissue from the Stanley Foundation brain collection, Bethesda, MD. This collection consists of 15 schizophrenia, 15 bipolar disorder, 15 major depression, and 15 control individuals, all 60 brains being well-matched on conventional parameters, with antipsychotic drug exposure in the schizophrenia and bipolar disorder groups. Significant up-regulation of apo L1 gene expression in schizophrenia was confirmed. Using quantitative PCR, expression profiles of other members of the apo L family (apo L2-L6) were investigated, showing that apo L2 and L4 were highly significantly up-regulated in schizophrenia. Results were then confirmed in an independent set of 20 schizophrenia and 20 control brains from Japan and New Zealand. Apo L proteins belong to the group of high density lipoproteins, with all six apo L genes located in close proximity to each other on chromosome 22q12, a confirmed high-susceptibility locus for schizophrenia and close to the region associated with velocardiofacial syndrome that includes symptoms of schizophrenia. PMID- 11930016 TI - Mapping the evolution of regional atrophy in Alzheimer's disease: unbiased analysis of fluid-registered serial MRI. AB - Alzheimer's disease (AD) is characterized by progressive cerebral atrophy, which may be assessed by using volumetric MRI. We describe a voxel-based analysis of nonlinear-registered serial MRI to demonstrate the most statistically significant (P < 0.001) regions of change at different stages of the disease. We compared presymptomatic (n = 4), mild (n = 10), and moderately affected (n = 12) patients with early- and late-onset AD, with age- and sex-matched controls, and demonstrated increasing global atrophy with advancing disease. Significantly increased rates of hippocampal atrophy were seen in presymptomatic and mildly affected patients. There was a shift in the distribution of temporal lobe atrophy with advancing disease; the inferolateral regions of the temporal lobes showed the most significantly increased rates of atrophy by the time the patients were mildly or moderately affected. Significantly increased rates of medial parietal lobe atrophy were seen at all stages, with frontal lobe involvement occurring later in the disease. Our results suggest that the sites showing the most significant rates of atrophy alter as the disease advances, and that regional atrophy is already occurring before the onset of symptoms. This technique provides insights into the natural history of AD, and may be a valuable tool in assessing the efficacy of disease-modifying treatments, especially if these treatments were to have region-specific effects. PMID- 11930017 TI - Cloning and characterization of a cAMP-specific phosphodiesterase (TbPDE2B) from Trypanosoma brucei. AB - Here we report the cloning, expression, and characterization of a cAMP-specific phosphodiesterase (PDE) from Trypanosoma brucei (TbPDE2B). Using a bioinformatic approach, two different expressed sequence tag clones were identified and used to isolate the complete sequence of two identical PDE genes arranged in tandem. Each gene consists of 2,793 bases that predict a protein of 930 aa with a molecular mass of 103.2 kDa. Two GAF (for cGMP binding and stimulated PDEs, Anabaena adenylyl cyclases, and Escherichia coli FhlA) domains, similar to those contained in many signaling molecules including mammalian PDE2, PDE5, PDE6, PDE10, and PDE11, were located N-terminal to a consensus PDE catalytic domain. The catalytic domain is homologous to the catalytic domain of all 11 mammalian PDEs, the Dictyostelium discoideum RegA, and a probable PDE from Caenorhabditis elegans. It is most similar to the T. brucei PDE2A (89% identity). TbPDE2B has substrate specificity for cAMP with a K(m) of 2.4 microM. cGMP is not hydrolyzed by TbPDE2B nor does this cyclic nucleotide modulate cAMP PDE activity. The nonselective PDE inhibitors 3-isobutyl-1-methylxanthine, papaverine and pentoxifyline are poor inhibitors of TbPDE2B. Similarly, PDE inhibitors selective for the mammalian PDE families 2, 3, 5, and 6 (erythro-9-[3-(2-hydroxynonyl)]-adenine, enoximone, zaprinast, and sildenafil) were also unable to inhibit this enzyme. However, dipyridamole was a reasonably good inhibitor of this enzyme with an IC50 of 27 microM. cAMP plays key roles in cell growth and differentiation in this parasite, and PDEs are responsible for the hydrolysis of this important second messenger. Therefore, parasite PDEs, including this one, have the potential to be attractive targets for selective drug design. PMID- 11930018 TI - Structural requirement of bilin chromophore for the photosensory specificity of phytochromes A and B. AB - Phytochromes are an important class of chromoproteins that regulate many cellular and developmental responses to light in plants. The model plant species Arabidopsis thaliana possesses five phytochromes, which mediate distinct and overlapping responses to light. Photobiological analyses have established that, under continuous irradiation, phytochrome A is primarily responsible for plant's sensitivity to far-red light, whereas the other phytochromes respond mainly to red light. The present study reports that the far-red light sensitivity of phytochrome A depends on the structure of the linear tetrapyrrole (bilin) prosthetic group. By reconstitution of holophytochrome in vivo through feeding various synthetic bilins to chromophore-deficient mutants of Arabidopsis, the requirement for a double bond on the bilin D-ring for rescuing phytochrome A function has been established. In contrast, we show that phytochrome B function can be rescued with various bilin analogs with saturated D-ring substituents. PMID- 11930019 TI - GCR1, the putative Arabidopsis G protein-coupled receptor gene is cell cycle regulated, and its overexpression abolishes seed dormancy and shortens time to flowering. AB - Although signaling through heterotrimeric G proteins has been extensively studied in eukaryotes, there is little information about this important signaling pathway in plants. We observed that expression of GCR1, the gene encoding the only known (but still putative) G protein-coupled receptor of Arabidopsis thaliana, is modulated during the cell cycle and during plant development. Overexpression of GCR1 in tobacco (Nicotiana tabacum) BY-2 cells caused an increase in thymidine incorporation and in the mitotic index of aphidicolin synchronized cells. Overexpression of GCR1 in Arabidopsis caused two remarkable phenotypes: seed dormancy was abolished and time to flowering was reduced. Molecular markers of these two developmental processes (phosphatase PP2A and MYB65 in germination; LFY during flowering) were up-regulated in GCR1 overexpressors. These data are consistent with the hypothesis that GCR1 may be a regulator of the cell cycle and that this regulation underlies the developmental changes observed in the GCR1 transformants. PMID- 11930020 TI - Chaos in learning a simple two-person game. AB - We investigate the problem of learning to play the game of rock-paper-scissors. Each player attempts to improve her/his average score by adjusting the frequency of the three possible responses, using reinforcement learning. For the zero sum game the learning process displays Hamiltonian chaos. Thus, the learning trajectory can be simple or complex, depending on initial conditions. We also investigate the non-zero sum case and show that it can give rise to chaotic transients. This is, to our knowledge, the first demonstration of Hamiltonian chaos in learning a basic two-person game, extending earlier findings of chaotic attractors in dissipative systems. As we argue here, chaos provides an important self-consistency condition for determining when players will learn to behave as though they were fully rational. That chaos can occur in learning a simple game indicates one should use caution in assuming real people will learn to play a game according to a Nash equilibrium strategy. PMID- 11930021 TI - Pulmonary arteriovenous malformation. AB - Pulmonary arteriovenous malformations (PAVM) are rare pulmonary vascular anomalies. Although most patients are asymptomatic, PAVMs can cause dyspnoea from right-to-left shunt. Because of paradoxical emboli, various central nervous system complications have been described including stroke and brain abscess. There is a strong association between PAVM and hereditary haemorrhagic telangiectasia. Chest radiography and contrast enhanced computed tomography are essential initial diagnostic tools but pulmonary angiography is the gold standard. Contrast echocardiography is useful for diagnosis and monitoring after treatment. Most patients should be treated. Therapeutic options include angiographic embolisation with metal coil or balloon occlusion and surgical excision. PMID- 11930022 TI - Enteral feeding. Nasogastric, nasojejunal, percutaneous endoscopic gastrostomy, or jejunostomy: its indications and limitations. AB - The following article is intended to provide a review of the current state of enteral feeding; a rapidly changing and developing field. It covers the type of feed, the routes of access, and the problems that can occur with enteral feeding. PMID- 11930023 TI - Japanese viral encephalitis. AB - One of the leading causes of acute encephalopathy in children in the tropics is Japanese encephalitis (JE). Transmitted by the culex mosquito, this neurotropic virus predominately affects the thalamus, anterior horns of the spinal cord, cerebral cortex, and cerebellum. It mainly affects children <15 years and is mostly asymptomatic. The occasional symptomatic child typically presents with a neurological syndrome characterised by altered sensorium, seizures, and features of intracranial hypertension. Aetiological diagnosis is based on virus isolation or demonstration of virus specific antigen or antibodies in the cerebrospinal fluid/blood. Though no antiviral drug is available against JE, effective supportive management can improve the outcome. Control of JE involves efficient vector control and appropriate use of vaccines. PMID- 11930024 TI - Mechanisms of bacterial pathogenicity. AB - Pathogenic bacteria utilise a number of mechanisms to cause disease in human hosts. Bacterial pathogens express a wide range of molecules that bind host cell targets to facilitate a variety of different host responses. The molecular strategies used by bacteria to interact with the host can be unique to specific pathogens or conserved across several different species. A key to fighting bacterial disease is the identification and characterisation of all these different strategies. The availability of complete genome sequences for several bacterial pathogens coupled with bioinformatics will lead to significant advances toward this goal. PMID- 11930025 TI - Role of the human papilloma virus in the development of cervical intraepithelial neoplasia and malignancy. AB - Human papilloma virus (HPV) is a public health problem as a sexually transmitted disease and as a critical factor in the pathogenesis of various cancers. The clinical manifestations, epidemiology, and virology that are critical to understanding the process of cervical dysplasia and neoplasia are reviewed. A discussion of the cervical transformation zone and the classification of cervical dysplasia and neoplasia leads into the importance of the Papanicolaou smear in prevention of potentially devastating sequelae of this virus. The role of the immune system in the progression of the disease and how it relates to vaccines, as well as treatment and prevention of HPV, are reviewed. PMID- 11930026 TI - Are single mothers in Britain failing to monitor their oral health? AB - OBJECTIVES: This study was designed to identify association between self reported dental attendance patterns and family structure in the UK. DESIGN: A national study involving 666 women with dependent children. SETTING: Home interviews were undertaken exploring time and reason for last dental visit. In addition, numerous sociodemographic and service related characteristics were collected. RESULTS: Bivariate analysis identified that family structure was associated with respondents' self reported dental attendance patterns: marital status (p<0.01), number of children (p<0.05), and age of children (p<0.05). When the combined effects of age, family structure, income, educational attainment, working status, and service factors (difficulty obtaining a NHS dentist and time taken to get an appointment) on dental attendance were explored, family structure emerged as a very important predicator of service use. Notably, young (age 16-34) single mothers and those with more than two children were less likely to have attended the dentist within the past year for reasons other than a dental emergency compared with older (age 35 or more), mothers from a two parent family and those with one or two children. CONCLUSION: Family structure is associated with self reported dental attendance patterns. Young single mothers with more than two children may be failing to monitor their oral health appropriately. PMID- 11930027 TI - Breath analysis to detect recent exposure to carbon monoxide. AB - OBJECTIVES: To determine the normal range for carbon monoxide concentrations in the exhaled breath of subjects in the emergency department and to develop a protocol for the use of a breath analyser to detect abnormal carbon monoxide exposure. METHODS: A hand held breath analyser was used to measure end expiratory carbon monoxide concentrations in 382 consenting subjects. Questionnaire data were collected to assess the effect of common sources of carbon monoxide exposure on breath carbon monoxide levels. Smokers were used as a carbon monoxide exposed group for comparison with non-smokers. RESULTS: The range of carbon monoxide concentrations obtained in the non-smoking group was 0-6 ppm and in the smoking group was 1-68 ppm. Smokers had a mean breath carbon monoxide concentration of 16.4 ppm and non-smokers had a mean of 1.26 ppm (95% confidence interval (CI) for difference 13.6 to 16.8 ppm). Male sex and frequent motor vehicle use were associated with slightly higher carbon monoxide concentrations (by 0.40, 95% CI 0.18 to 0.63 ppm, and 0.38, 95% CI 0.13 to 0.63 ppm, respectively) in the non smoking group. Mean breath carbon monoxide concentrations increased in direct proportion to the number of cigarettes smoked (p<0.001) and there was a negative correlation between carbon monoxide and time since last smoking a cigarette (p<0.001). Altogether 23% of smokers had breath carbon monoxide concentrations in the range 1-6 ppm. CONCLUSIONS: Breath analysis was rapid and results correlated well with carbon monoxide exposure. In this population subjects with breath carbon monoxide concentrations greater than 6 ppm should be assessed for the risk of carbon monoxide poisoning. However even carbon monoxide concentrations less than 6 ppm do not exclude carbon monoxide poisoning within the last 24 hours. PMID- 11930028 TI - Prognosis of Japanese encephalitis patients with dystonia compared to those with parkinsonian features only. AB - OBJECTIVES: A number of movement disorders have been reported in Japanese encephalitis (JE). The prognostic significance of these movement disorders, however, has not been evaluated. The present study reports the prognostic significance of parkinsonian features and dystonia in JE. PATIENTS AND METHODS: During 1992 and 1998, 50 JE patients were managed; 35 of them developed movement disorders (the study group). The diagnosis of JE was based on clinical, radiological, and serological criteria. Parkinsonian features were rated by the unified Parkinson's disease rating scale and dystonia by the dystonia rating scale. The patients with parkinsonian features only were classified into group I and those with additional dystonia or dyskinesia into group II. The outcome was defined at the end of three months into poor, partial, and complete recovery depending on how the patients coped with daily living activities. RESULTS: The patients' ages ranged from 2 to 64 years and 11 were females. The admission mean Glasgow coma scale score was 6.9 (range 4-13). The movement disorders were noted after 1-4 weeks of ictus. There were 16 patients in group I and 19 in group II. The parkinsonian features were more pronounced in group II than in group I. At three months of follow up, fewer patients had parkinsonian features in group I than group II. Hypophonia, however, persisted in 12 patients in group I and 16 in group II until the three month follow up. In group II, the mean dystonia score was 3.2 which regressed to 1.8 at three months. Tremor was present in five patients in groups I and eight in group II. Cranial computed tomography was abnormal in six and magnetic resonance imaging abnormal in 15 patients in group I and in nine and 12 patients respectively in group II. The thalamus was most frequently involved (11 patients in each group), basal ganglia (four in group I and six in group II), and midbrain (six in group I and one in group II). Group II patients had poorer recovery compared with group I. In group I, at the end of three months functional recovery was complete in 10, partial in two, and poor in three patients. In group II, four patients had complete, seven partial, and eight poor recovery. CONCLUSION: JE results in a transient form of parkinsonian syndrome, which is associated with a lower frequency of tremor and prominent hypophonia. The presence of dystonia suggests more severe illness and poorer prognosis. PMID- 11930029 TI - Osteoporosis prophylaxis during corticosteroid treatment: failure to prescribe. AB - AIMS: To investigate prescribing patterns to prevent steroid induced osteoporosis. To compare prophylactic prescribing with National Osteoporosis Society (NOS) guidelines. METHOD: All patients (n=92) taking oral corticosteroids admitted to general medical wards at a district general hospital were prospectively investigated over a nine month period. RESULTS: Variations from recommended management were revealed. Altogether 64.7% of all inpatients who qualified for prophylaxis for steroid induced osteoporosis were not provided with any suitable agent. It was also found that 21.6% of those who qualified for treatment received a bisphosphonate, the only treatment currently licensed for preventing steroid induced osteoporosis. Of those prescribed prophylactic treatment, a bisphosphonate was selected for 39.3%, hormone replacement therapy was given to 25.0%, and 35.7% received treatment that is not recommended in NOS guidelines. CONCLUSION: This study revealed substantial variations from NOS guidelines. It is suggested that osteoporosis prophylaxis during steroid treatment is promoted by local hospital guidelines, hospital and community pharmacists, audit, and general practitioners. PMID- 11930030 TI - Troponin T elevation in lobar lung disease. AB - Two patients with a rise in cardiac troponin T (cTnT) concentrations during the course of lobar pneumonia, without any evidence of acute coronary syndrome or renal failure, are presented. The increase in cTnT concentration is considered a highly sensitive marker of cardiac injury, although it may also rise in other conditions and as a result of lobar pneumonia. Thus, this effect should be considered when the possibility of acute coronary syndrome in such patients is addressed. PMID- 11930032 TI - A proliferating pimple. PMID- 11930033 TI - An interesting case of thirst and polyuria. PMID- 11930031 TI - Transient ischaemic attacks mimicking focal motor seizures. AB - Limb shaking is an under-recognised form of transient ischaemic attack (TIA), which can easily be confused with focal motor seizures. However, it is important to distinguish limb shaking TIAs and focal seizures, as patients with this form of TIA almost invariably have severe carotid occlusive disease and are at high risk of stroke. A patient with limb shaking TIAs is presented in whom the diagnosis was missed. PMID- 11930034 TI - An episodic eruption. PMID- 11930035 TI - A 15 year old girl with fever, jaundice, haemolysis, and sudden clinical deterioration. PMID- 11930040 TI - The 21st century: the age of screening. PMID- 11930041 TI - The ALARA concept in pediatric CT: myth or reality? PMID- 11930042 TI - Forensic radiology: response to the Pentagon Attack on September 11, 2001. PMID- 11930043 TI - A suggestion: look at the images first, before you read the history. PMID- 11930044 TI - Intracranial mass lesions: dynamic contrast-enhanced susceptibility-weighted echo planar perfusion MR imaging. AB - Dynamic contrast agent-enhanced perfusion magnetic resonance (MR) imaging provides physiologic information that complements the anatomic information available with conventional MR imaging. Analysis of dynamic data from perfusion MR imaging, based on tracer kinetic theory, yields quantitative estimates of cerebral blood volume that reflect the underlying microvasculature and angiogenesis. Perfusion MR imaging is a fast and robust imaging technique that is increasingly used as a research tool to help evaluate and understand intracranial disease processes and as a clinical tool to help diagnose, manage, and understand intracranial mass lesions. With the increasing number of applications of perfusion MR imaging, it is important to understand the principles underlying the technique. In this review, the essential underlying physics and methods of dynamic contrast-enhanced susceptibility-weighted echo-planar perfusion MR imaging are described. The clinical applications of cerebral blood volume maps obtained with perfusion MR imaging in the differential diagnosis of intracranial mass lesions, as well as the pitfalls and limitations of the technique, are discussed. Emphasis is on the clinical role of perfusion MR imaging in providing insight into the underlying pathophysiology of cerebral microcirculation. PMID- 11930045 TI - MR anatomy of the proximal cisternal segment of the trochlear nerve: neurovascular relationships and landmarks. AB - PURPOSE: To assess the anatomic features and vascular relationships of the proximal portion of the cisternal segment of the trochlear nerve. MATERIALS AND METHODS: In 30 subjects (60 nerves) and in one patient with right superior oblique myokymia (SOM), the anatomy of the trochlear nerve was depicted with three-dimensional (3D) Fourier transformation constructive interference in steady state (CISS) magnetic resonance (MR) imaging, whereas the adjacent vessels were detected with 3D time-of-flight (TOF) MR imaging before and after gadopentetate dimeglumine administration. The images were evaluated with respect to the identification of the trochlear nerve, the distance between the point of exit (PE) and the midline, the visualized length, the vascular relationships, and the distance between the PE and the point of neurovascular contact. RESULTS: 3D CISS MR imaging depicted the proximal cisternal segment of the trochlear nerve in the transverse, sagittal, and coronal planes in 57 (95%), 51 (85%), and 48 (80%) of 60 nerves, respectively. The distance from the midline to the PE was 3-9 mm, and the maximum visualized length of the trochlear nerve was 1-14 mm. An arterial trochlear neurovascular contact was seen at the root exit zone (REZ) in eight (14%) nerves and at a mean distance of 3.4 mm distal to the PE in 29 nerves (51%). The patient with SOM had arterial-trochlear neurovascular contact at the REZ. CONCLUSION: Use of 3D CISS sequences and 3D TOF sequences with or without gadopentetate dimeglumine enables accurate identification of the proximal cisternal segment of the trochlear nerve and its neurovascular relationships. PMID- 11930046 TI - Demonstration of the artery of Adamkiewicz at multi- detector row helical CT. AB - PURPOSE: To assess the ability of multi-detector row helical computed tomography (CT) to depict the artery of Adamkiewicz. MATERIALS AND METHODS: Seventy patients with vascular diseases underwent multi-detector row helical CT of the entire aorta and iliac arteries. The artery of Adamkiewicz was examined on multiplanar and curved planar reformation images and on cine-mode displays. The visualization of the artery of Adamkiewicz, as well as its branching level and side of origin, was investigated. RESULTS: In 63 (90%) of the 70 patients, at least a single artery of Adamkiewicz was clearly visualized from the intervertebral foramen to the hairpin-shaped union with the anterior spinal artery. Two arteries of Adamkiewicz were identified in 15 (24%) of 63 patients. Fifty-five arteries of Adamkiewicz (71%) originated from the left side. Seventy-two (92%) originated between T8 and L1. Neither the intercostal vein nor the posterior spinal vein were visualized in 57 of 63 patients. Continuity of the entire length, starting from the stem of the intercostal or lumbar artery and proceeding to the artery of Adamkiewicz and finally to the anterior spinal artery, was traceable on cine-mode displays or on curved planar reformation images in 20 of 63 patients. CONCLUSION: Multi-detector row helical CT depicts the artery of Adamkiewicz in a high percentage of patients. PMID- 11930047 TI - Differentiation of multiple sclerosis from other inflammatory disorders and cerebrovascular disease: value of spinal MR imaging. AB - PURPOSE: To determine the value of magnetic resonance (MR) imaging in the spinal cord to differentiate multiple sclerosis (MS) from other inflammatory disorders and cerebrovascular diseases (together, other neurologic disease [OND]). MATERIALS AND METHODS: The study population included 66 patients with OND and 25 patients with MS, who were matched for age, sex, and symptom duration or severity. Brain MR imaging included gadolinium-enhanced T1-weighted and dual-echo T2-weighted spin-echo sequences to assess the number, size, and appearance of lesions, contrast enhancement, and compatibility with diagnostic criteria for MS. Spinal cord MR imaging included cardiac-triggered gadolinium-enhanced sagittal T1 weighted spin-echo and dual-echo T2-weighted sequences to assess the general appearance (normal, focal lesion, diffuse abnormality) and number or size of focal lesions. Images obtained in MS and OND patients were compared. Specificity, sensitivity, accuracy, and positive and negative predictive values with MR images were calculated. RESULTS: Brain images were abnormal in all MS patients and in 65% of OND patients. Abnormal cord images were found in 92% of MS and 6% of OND patients. The combination of brain and spinal cord images increased accuracy of diagnosis compared with use of brain images alone. CONCLUSION: In contrast to MS, cord lesions are very uncommon in OND. This finding can help differentiate these disorders. PMID- 11930048 TI - Improvement of treatment plans developed with intensity-modulated radiation therapy for concave-shaped head and neck tumors. AB - PURPOSE: To improve dose conformity and normal tissue sparing in patients with concave-shaped head and neck cancers by using tomotherapy and static step-and shoot intensity-modulated radiation therapy (IMRT) and by comparing results with those of three-dimensional (3D) conformal radiation therapy (CRT) and two dimensional (2D) radiation therapy. MATERIALS AND METHODS: Treatment planning in 10 patients with concave-shaped head and neck tumors was performed by using tomotherapy and step-and-shoot IMRT, 3D CRT, and 2D techniques. IMRT plans were modified by placing "virtual critical structures" in regions outside the target where hot spots occurred. These modified plans were used for comparison because they provided better dose conformity. Critical structures were the spinal cord, the parotid glands, and the mandible. Comparisons were performed by means of dose volume histograms, clinical target volume (CTV), target covered by 95% isodose (D(95%)), dose received by 5% of the critical structure volume (D(5%)), maximum dose, mean dose, and normal tissue complication probability for critical structures. RESULTS: Original IMRT plans showed more conformal dose distributions than those in 3D CRT and 2D plans. However, hot spots developed in the posterior and anterior neck. Introduction of virtual critical structures in IMRT plans resulted in removal of these hot spots without affecting target coverage. Modified IMRT plans also demonstrated better CTV coverage than that in 3D CRT and 2D plans. The average D(95%) was 97.3% with tomotherapy, 97.1% with step-and shoot IMRT, 84.7% with 3D CRT, and 69.4% with 2D techniques. D(5%) for the spinal cord changed from approximately 45 Gy with 3D plans and 46 Gy with 2D plans to approximately 28 Gy with IMRT. CONCLUSION: IMRT demonstrated better target coverage and sparing of critical structures than that of 3D CRT and 2D techniques. Use of virtual critical structures resulted in removal of hot spots around the spinal cord. PMID- 11930049 TI - Transient hypoxia-ischemia in rats: changes in diffusion-sensitive MR imaging findings, extracellular space, and Na+-K+ -adenosine triphosphatase and cytochrome oxidase activity. AB - PURPOSE: To investigate the correlation between diffusion-weighted (DW) magnetic resonance (MR) image changes with alterations in extracellular volume and changes in cytochrome oxidase and Na(+)-K(+)-adenosine triphosphatase (ATPase) activity at various times during and after cerebral hypoxia-ischemia in neonatal and juvenile rats. MATERIALS AND METHODS: One- and 4-week-old rats were randomly assigned to control or transient cerebral hypoxia-ischemia (ie, right carotid artery occlusion plus exposure to 8% oxygen) groups. Hypoxic-ischemic changes compared with normal ipsilateral brain tissue on DW images and the apparent diffusion coefficient of water were measured during and at 1 and 24 hours after hypoxia-ischemia ended. Hypoxic-ischemic changes in extracellular space and ipsilateral versus contralateral differences in Na(+)-K(+)-ATPase and cytochrome oxidase activity were measured. RESULTS: Hyperintensities on DW images obtained during hypoxia-ischemia correlated well (P <.05) with extracellular space reductions, which occurred 15 minutes earlier in the brains of 4-week-old rats than in the brains of 1-week-old rats. Similarly, within 1 hour after hypoxia ischemia ended, DW image and extracellular space changes normalized. In contrast, Na(+)-K(+)-ATPase and cytochrome oxidase activity decreased in some regions during hypoxia-ischemia and remained reduced 1 hour after the end of hypoxia ischemia. Twenty-four hours after signal intensity normalization, hyperintense areas reappeared on DW images, and Na(+)-K(+)-ATPase and cytochrome oxidase activity remained decreased. CONCLUSION: Signal intensity alterations with diffusion-sensitive MR imaging during and after transient hypoxia-ischemia are closely associated with a corresponding shrinkage and reexpansion of the extracellular space, irrespective of age. Mechanisms other than Na(+)-K(+)-ATPase changes may induce the early cell volume changes detected with diffusion sensitive MR imaging. PMID- 11930050 TI - Noninvasive measurement of extraction fraction and single-kidney glomerular filtration rate with MR imaging in swine with surgically created renal arterial stenoses. AB - PURPOSE: To test whether magnetic resonance (MR) imaging enables accurate measurement of extraction fraction (EF) in swine with unilateral renal ischemia and to evaluate effects of renal arterial stenosis on EF and single-kidney glomerular filtration rate. MATERIALS AND METHODS: High-grade unilateral renal arterial stenoses were surgically created in eight pigs. Direct measurements of renal venous and arterial inulin concentration provided reference standard estimates of single-kidney EF. Pigs were imaged with a 1.5-T imager to estimate EF, renal blood flow, and glomerular filtration rate. A breath-hold inversion recovery spiral sequence was used to measure T1 of blood in the infrarenal inferior vena cava and renal veins after intravenous administration of gadopentetate dimeglumine, and these data were used to calculate EF. Cine-phase contrast material-enhanced imaging of the renal arteries provided quantitative renal blood flow measurements. Bilateral single-kidney glomerular filtration rate was then determined: glomerular filtration rate = renal blood flow x (1 - hematocrit level) x EF. RESULTS: A statistically significant linear correlation was found between EF, as determined with MR imaging, and inulin (r = 0.77). As compared with kidneys without renal arterial stenosis, kidneys with renal arterial stenosis showed 50% (0.14/0.28) EF reduction (P <.01) and 59% glomerular filtration rate reduction (P <.01). CONCLUSION: MR imaging shows promise for in vivo measurement of EF and glomerular filtration rate, which may be useful in assessing the clinical importance of renal arterial stenosis. PMID- 11930051 TI - Covered, retrievable, expandable urethral nitinol stent: feasibility study in dogs. AB - PURPOSE: To evaluate the feasibility of using a retrievable urethral stent and to evaluate whether granulation tissue resolves after stent removal in a canine urethra. MATERIALS AND METHODS: Polyurethane-covered retrievable 0.10-mm-thick (n = 11) or 0.15-mm-thick (n = 10) nitinol wire stents were placed in the urethras of 20 dogs. In one dog, a second stent was placed in the urethra because of complete migration of the first stent into the urinary bladder. The stents were removed with retrieval hook wires 4 weeks (n = 10) and 8 weeks (n = 10) after placement. Fourteen dogs were sacrificed just after stent removal, and the other six dogs were sacrificed 2 weeks after stent removal. Information concerning procedure success, stent migration, and tissue response was obtained. RESULTS: Stent placement was technically successful in all dogs. Follow-up urethrograms showed partial (n = 4) or complete (n = 1) stent migration. Stent removal failed in two dogs due to partial or complete migration. Granulation tissue was observed at both ends of the stent in 17 dogs. Urethrograms and urethral specimens obtained 2 weeks after stent removal showed diminished granulation tissue and decreased thickness of the papillary projections of the epithelium compared with results obtained immediately after stent removal. CONCLUSION: Although some design modifications are necessary to reduce current complications, the polyurethane-covered retrievable nitinol stent seems feasible for use in the urethra. Stent-induced granulation tissue formation improved after stent removal. PMID- 11930052 TI - Prostate cancer tumor volume: measurement with endorectal MR and MR spectroscopic imaging. AB - PURPOSE: To determine accuracy of magnetic resonance (MR) and three-dimensional (3D) MR spectroscopic imaging in prostate cancer tumor volume measurement. MATERIALS AND METHODS: Endorectal MR and 3D MR spectroscopic imaging were performed in 37 patients before radical prostatectomy. Two independent readers recorded peripheral zone tumor nodule location and volume. Results were analyzed with step-section histopathologic tumor localization and volume measurement as the standard. Accuracy of tumor volume measurement was assessed with the Pearson correlation coefficient. P values were calculated with a random effects model. Bland-Altman regression analysis was used to evaluate systematic bias between tumor volumes measured with MR imaging and true tumor volumes. Analyses were performed for all nodules and nodules greater than 0.50 cm(3). RESULTS: Mean volume of peripheral zone tumor nodules (n = 51) was 0.79 cm(3) (range, 0.02-3.70 cm(3)). Two readers detected 20 (65%) and 23 (74%) of 31 peripheral zone tumor nodules greater than 0.50 cm(3). For these nodules, measurements of tumor volume with MR imaging, 3D MR spectroscopic imaging, and a combination of both were all positively correlated with histopathologic volume (Pearson correlation coefficients of 0.49, 0.59, and 0.55, respectively); only measurements with 3D MR spectroscopic imaging and a combination of MR and 3D MR spectroscopic imaging demonstrated statistical significance (P <.05). Tumor volume estimation with all three methods was more accurate for higher tumor volumes. CONCLUSION: Addition of 3D MR spectroscopic imaging to MR imaging increases overall accuracy of prostate cancer tumor volume measurement, although measurement variability limits consistent quantitative tumor volume estimation, particularly for small tumors. PMID- 11930053 TI - Patients with acute flank pain: comparison of MR urography with unenhanced helical CT. AB - PURPOSE: To compare unenhanced helical computed tomography (CT) and magnetic resonance (MR) urography, by using T2-weighted and contrast material-enhanced T1 weighted imaging to examine patients with acute flank pain, with reference to excretory urography and final clinical diagnosis. MATERIALS AND METHODS: Forty nine patients underwent CT, MR urography (with T2-weighted and gadopentetate dimeglumine-enhanced T1-weighted sequences), and excretory urography. CT and MR urographic findings were evaluated separately and independently by two radiologists each (CT, observers A and B; MR urography, observers C and D) for the presence, cause, level, and degree of obstruction. The final conclusive diagnosis was based on the combination of excretory urographic, clinical, and interventional results. RESULTS: At final diagnosis, 32 (65%) patients were found to have ureteral stones causing unilateral obstruction. In ureteral stone detection, the sensitivity and specificity of CT were 90.6% (29 of 32 patients) and 100.0% (17 of 17 patients), respectively (observer A) and 90.6% (29 of 32 patients) and 94.1% (16 of 17 patients), respectively (observer B), while those of MR urography were 93.8% (30 of 32 patients) and 100.0% (17 of 17 patients), respectively (observer C) and 100.0% (32 of 32 patients) and 100.0% (17 of 17 patients), respectively (observer D). Spearman correlation coefficients for stone size at CT were 0.76 (P <.001) and 0.75 (P <.001) and at MR urography, 0.49 (P =.005) and 0.51 (P =.004). CONCLUSION: In routine clinical practice, CT is the modality of choice in the evaluation of patients with acute flank pain. MR urography is an accurate and suitable alternative imaging technique in selected patients. PMID- 11930054 TI - Cutting balloon percutaneous transluminal angioplasty for salvage of lower limb arterial bypass grafts: feasibility. AB - PURPOSE: To evaluate the feasibility of cutting balloon percutaneous transluminal angioplasty (PTA) for treatment of neointimal hyperplasia in peripheral arterial bypass grafts. MATERIALS AND METHODS: Fifteen consecutive patients (six women, nine men; age range, 57-89 years; mean age, 71 years) were treated with cutting balloon PTA for 16 anastomotic stenoses after infrainguinal bypass (prosthetic grafts, seven patients; prosthetic-vein composite grafts, two; venous grafts, five; and ileofemoral stent-graft, one). Cutting balloon PTA was followed by conventional PTA to improve anastomotic diameter. Patients with stenotic vein grafts underwent cutting balloon PTA after failed conventional PTA; the other patients were treated primarily with cutting balloon PTA. Criteria for success were a lumen diameter improvement of greater than 50% or residual stenosis of 20% or less. Follow-up was performed with color duplex ultrasonographic surveillance. Patency rates and durations were calculated with Kaplan-Meier survival curves and log-rank statistics. RESULTS: Attempted conventional PTA (n = 6) prior to cutting balloon PTA was unsuccessful. Cutting balloon PTA was technically successful in 15 (94%) of 16 lesions, without clinical complications. Two local restenoses and one graft occlusion occurred between 5 and 7 months. The cumulative 6-month primary and secondary graft patency rates were 84% and 92%, respectively. At 12 and 18 months, they were 67% (95% CI: 0.34, 0.86) and 83% (95% CI: 0.48, 0.96), respectively; mean follow-up was 10.0 months. CONCLUSION: Cutting balloon PTA proved feasible for treatment of resistant peripheral arterial bypass graft stenosis, commonly caused by neointimal hyperplasia, with excellent technical success. Short-term patency with this technique appears to be superior to that with conventional PTA, and it compares well with patency of atherectomy for salvage of infrainguinal bypass grafts. PMID- 11930055 TI - CT-guided transthoracic percutaneous ethanol injection for hepatocellular carcinoma not detectable with US. AB - PURPOSE: To evaluate the safety and effectiveness of computed tomography (CT) guided percutaneous ethanol injection (PEI) for the treatment of hepatocellular carcinoma (HCC) not detectable with ultrasonography (US). MATERIALS AND METHODS: Between April 1994 and January 2001, 51 patients with 57 HCC nodules not detectable with US underwent CT-guided transthoracic PEI. Complications associated with the transthoracic approach, effectiveness of transthoracic PEI, and prognosis of the patients were evaluated. RESULTS: Seventy-one PEI sessions were performed for 57 nodules. Complications included pneumothorax in 21 sessions (30%) for 19 nodules (33%), moderate pleural effusion in four sessions (6%) for four nodules (7%), and hemoptysis in three sessions (4%) for two nodules (4%). A chest tube was required for pneumothorax in five sessions (7%) for five nodules (9%), and pleural effusion drainage was performed in two sessions (3%) for two nodules (4%). Apparent tumor necrosis was noted at CT in 51 nodules (89%). During follow-up (range, 3 months to 5(1/2) years; mean, 29 months +/- 18 [SD]), local recurrence was seen in seven nodules (12%), three of which received repeat treatment with transthoracic PEI. Twenty-six patients survived, and 25 patients died of multiple tumors, hepatic failure, or rupture of esophageal varices. CONCLUSION: Transthoracic PEI seems to be relatively safe and effective for the treatment of HCC not detectable with US. PMID- 11930056 TI - Percutaneous vertebroplasty for severe osteoporotic vertebral body compression fractures. AB - PURPOSE: To determine the efficacy of percutaneous vertebroplasty in treating severe vertebral body compression fractures, or vertebra plana, in patients with osteoporosis. MATERIALS AND METHODS: In 155 patients, 310 percutaneous vertebroplasties were performed during 25 months and 15 days. Of these, 37 patients (27 women, 10 men; mean age, 73.6 years) underwent 48 vertebroplasties for severe osteoporotic vertebral body compression fractures. The fractures were defined as vertebrae that have collapsed to less than one-third of their original height. Imaging and clinical features were analyzed, including the extent of vertebral collapse, location of the involved vertebra, pattern of vertebral compression, volume of polymethylmethacrylate injected, vertebroplasty complications, and clinical outcome. RESULTS: Vertebral body collapse averaged 23% (range, 4.5%-33.0%) of the original height. Involved vertebrae were located from levels T5 to L5, with one-half affected at the thoracolumbar junction. Patterns of vertebral compression were divided into gibbus (31 of 48 or 65%), plana (13 of 48 or 27%), and H shape (four of 48 or 8%). The mean volume of the cement injected was 6.0 mL (range, 1.5-12.5 mL). Complications observed on radiographs included cement leakage to the adjacent disc (17 of 48 or 35%) and the paravertebral soft tissues (four of 48 or 8%). There were no major complications. At clinical follow-up (mean duration, 11 months and 3 days; range, 3-24 months), pain relief was complete in 14 (47%) of 30 patients, partial in 15 (50%), and unchanged in one (3%). No patient required surgery. CONCLUSION: Percutaneous vertebroplasty for severe osteoporotic vertebral body compression fractures is safe and effective and should not be withheld in this group of patients. PMID- 11930057 TI - Interventional musculoskeletal procedures performed by using MR imaging guidance with a vertically open MR unit: assessment of techniques and applicability. AB - PURPOSE: To evaluate the safety of and time required for a broad range of musculoskeletal interventional procedures performed by using magnetic resonance (MR) imaging guidance with a vertically open 0.5-T unit. MATERIALS AND METHODS: Sixty-three MR imaging-guided procedures were performed. A vertically open MR unit equipped with in-room display monitors allowed interactive freehand MR guidance predominantly with fast spin-echo and gradient-echo sequences. Each procedure was classified in terms of the anatomic location, procedure type, and tissue type involved. The procedures were evaluated for success of needle placement, adequacy of tissue sampling, total procedural time, needle time, number of needle passes, and complications. RESULTS: Procedures consisted of tissue sampling with core-needle (n = 6) or fine-needle aspiration (n = 20) biopsy, corticosteroid or contrast agent injection (n = 19), joint cyst aspiration (n = 7), and drainage (n = 11). Successful needle placement was achieved in all 63 cases. Cytologic and histologic tissue samples were sufficient for pathologic diagnosis in 24 of 26 cases. In two cases, complications occurred: transient local bleeding and a brief vasovagal episode. The mean total procedural time was 64.8 minutes; the mean needle time, 26.2 minutes; and the mean number of needle passes per patient, 1.6. CONCLUSION: With use of a vertically open MR unit, MR-guided interventional procedures involving bone, soft tissue, intervertebral disks, and joints are safe and sufficiently rapid for use in clinical practice. PMID- 11930058 TI - Sublabral foramen and buford complex: inferior extent of the unattached or absent labrum in 50 patients. AB - PURPOSE: To determine the inferiormost extent of the anterosuperior labral variants on conventional transverse MR images. MATERIALS AND METHODS: We reviewed transverse MR images in 50 consecutive patients with a sublabral foramen or Buford complex at arthroscopy. Images were randomly mixed with those of 58 patients with either a normal labrum (n = 20) or an anterior labral tear (n = 38) at arthroscopy. MR imaging was fat suppressed fast spin echo intermediate or T2 weighted (repetition time msec/effective echo time msec, 1,800-3,000/30-102). Two radiologists evaluated by means of consensus the anterior labrum while blinded to patient history and arthroscopic results. Transverse images obtained through the glenoid fossa were totalled to determine the midpoint. Sensitivity, specificity, and accuracy of MR for depicting a sublabral foramen or Buford complex were calculated along with 95% CIs, by using surgical findings as the reference standard. RESULTS: The sensitivity of MR for diagnosing a sublabral foramen or Buford complex was 0.94 (47 of 50 patients, 95% CI: 0.87, 1.00), specificity was 0.80 (16 of 20 patients, 95% CI: 0.62, 0.97), and accuracy was 0.90 (63 of 70 patients, 95% CI: 0.82, 0.97). The anterior labrum was abnormal on the first transverse section inferior to the midpoint in nine (18%) patients. The labrum was also abnormal on the second section below the midpoint in three (6%) patients. Because of the anterior tilt of the scapula, the midpoint was near the anterior glenoid notch at about the position between 2- and 3-o'clock. CONCLUSION: The labrum may be unattached or absent on the first two transverse images obtained below the midpoint. PMID- 11930059 TI - Recurrent glomus tumors of fingertips: MR imaging evaluation. AB - PURPOSE: To determine the magnetic resonance (MR) imaging findings in recurrent glomus tumors of the fingertips. MATERIALS AND METHODS: Twenty-four consecutive patients with recurrent pain after previous excision of a glomus tumor of the fingertip underwent MR imaging studies and surgery. T1-weighted spin-echo MR images were obtained in each patient before and after intravenous injection of contrast material; T2-weighted spin-echo and three-dimensional gradient-recalled echo images were also obtained. MR angiography was performed in four patients. Postsurgical histopathologic analysis revealed recurrent glomus tumors in 22 patients. Signal intensity, enhancement, and margins of the scar tissue and the recurrent tumors at MR were assessed. RESULTS: The postsurgical scars were depicted in 21 (88%) of 24 patients with all sequences but were best demonstrated on gradient-recalled echo MR images. Seven patients had undergone multiple surgical procedures and had extensive scar tissue and, in one case, a neuroma. In all patients, MR imaging revealed a nodule compatible with the diagnosis of a recurrent glomus tumor. In 13 (54%) of 24 patients, the nodule had typical features of a glomus tumor. In eight (33%) of 24 patients, the tumors had low signal intensity or isointensity compared with the nail bed on T2-weighted images. In six (25%) of 24 patients, the tumors had faint enhancement after intravenous gadolinium chelate administration. The margins of the tumors were blurred by scar tissue in nine of 24 cases. CONCLUSION: MR imaging can aid in the evaluation of recurrent glomus tumors. PMID- 11930060 TI - US evaluation of fetal growth: prediction of neonatal outcomes. AB - PURPOSE: To determine whether fetal growth measured at serial ultrasonographic (US) examinations can predict neonatal morbidity, independent of whether gestational age is known. MATERIALS AND METHODS: Women (n = 321) who had singleton pregnancies and underwent two or more second- or third-trimester obstetric US examinations were included in a retrospective cohort analysis. Inadequate fetal growth was defined as growth at or below the 10th percentile. The relative risk of each poor outcome was calculated for fetuses with inadequate growth, compared with the risk for fetuses with normal growth. RESULTS: Inadequate fetal growth was associated with 3.9 times the risk of a birth weight less than 2,500 g, 17.7 times the risk of a birth weight less than the 3rd percentile for gestational age, 2.3 times the risk of preterm birth, 2.6 times the risk of a long newborn hospital stay, and 3.6 times the risk of neonatal intensive care unit admission. After adjusting for confounding variables, including fetal weight, fetal growth remained a significant predictor of small birth size and poor outcomes. Inadequate growth predicted the risk of poor outcomes, even when gestational age was unknown. When inadequate growth was used to identify fetuses at risk, 21%-67% of neonates who were small at birth or had poor outcomes were identified at false-positive rates of only 5%-9%. For all outcomes, inadequate growth enabled identification of more fetuses with poor birth outcomes than low estimated fetal weight. CONCLUSION: Morbidity is significantly increased among fetuses who demonstrate less than expected growth. Growth between two US examinations can be used to estimate the risk of neonatal morbidity, even when gestational age is unknown. PMID- 11930062 TI - Case 45: cystic angiomatosis. PMID- 11930063 TI - Staging of neuroblastoma at imaging: report of the radiology diagnostic oncology group. AB - PURPOSE: To compare the accuracies of computed tomography (CT), magnetic resonance (MR) imaging, and bone scintigraphy in staging disease in patients with neuroblastoma. MATERIALS AND METHODS: Ninety-six children with newly diagnosed neuroblastoma were enrolled in a multicenter prospective cohort study. CT, MR, and bone scintigraphy were used to evaluate tumor stage. Sensitivity and specificity values and receiver operating characteristic (ROC) curve analyses were used to compare the accuracy of CT, MR, and scintigraphy for tumor staging. RESULTS: Eighty-eight patients were eligible for staging analysis, and 45 patients who underwent surgery at initial diagnosis were eligible for analysis of local tumor extent. CT and MR had sensitivities of 43% and 83%, respectively (P <.01), and specificities of 97% and 88%, respectively (P >.05), for detection of stage 4 disease. Areas under the ROC curves for CT and MR were 0.81 and 0.85, respectively (P =.06); that for scintigraphy was 0.83. Addition of scintigraphy to both CT and MR increased the areas under the ROC curves to 0.90 and 0.88, respectively. Accuracy of CT and MR for staging disease confined to the chest or abdomen (stages 1, 2, and 3) was poor. CONCLUSION: MR alone and CT and MR combined with bone scintigraphy enable the accurate detection of stage 4 disease. Both CT and MR perform poorly for local tumor staging. PMID- 11930064 TI - Defining normal upper airway motion in asymptomatic children during sleep by means of cine MR techniques. AB - PURPOSE: To define normal upper airway motion in asymptomatic children during sleep by means of dynamic cine magnetic resonance (MR) techniques. MATERIALS AND METHODS: In children referred for MR of the brain who required sedation, a sagittal midline cine MR sequence was performed. Motion of the nasopharynx, oropharynx, and hypopharynx was characterized as static patent, dynamic patent, intermittent collapse, or static collapsed; maximal diameter and greatest change in size were calculated in millimeters. Mouth position (open or closed) was determined. Parameters were compared with age (t test) and mouth position (Fisher exact test). RESULTS: In the 148 subjects (mean age, 3.4 years), the nasopharynx showed dynamic motion in 53 (36%). The oropharynx was most commonly collapsed in 98 (66%) of the patients. The hypopharynx showed dynamic motion in 72 (49%) of the patients and was never collapsed. Vertical motion was present in 77 (52%) of the patients. The mouth was open in 96 (65%) of the patients. There was a statistically significant correlation between mouth position and dynamic motion in the oropharynx (P =.006) and in the nasopharynx (P <.006) but not in the hypopharynx (P =.655). CONCLUSION: Dynamic changes in diameter were often seen in the nasopharynx and in the hypopharynx of asymptomatic sleeping children. However, collapse of the hypopharynx was not normally encountered. PMID- 11930065 TI - Mild intermittent asthma: CT assessment of bronchial cross-sectional area and lung attenuation at controlled lung volume. AB - PURPOSE: To evaluate, with thin-section computed tomography (CT), changes in bronchial cross-sectional area and lung attenuation induced by bronchial stimulation in patients with mild intermittent asthma, at a given lung volume monitored with pneumotachography. MATERIALS AND METHODS: Twelve patients with mild intermittent asthma who were nonsmokers (National Institutes of Health staging) and six nonsmoking healthy volunteers, age and sex ratio-matched, were examined by using helical thin-collimation CT at the level of basal bronchi at 65% of total lung capacity. Three sets of acquisitions were obtained: at baseline and after inhalation of methacholine and then salbutamol. Cross-sectional areas of bronchi greater than 4 mm(2) were segmented and calculated from CT images. Lung attenuation was measured in the anterior, lateral, and posterior areas of the right lung parenchyma. Gas trapping was evaluated by using thin-section CT at residual volume in six of the patients with asthma. Statistical analysis included two factors repeated-measurement analysis of variance and Mann-Whitney and Kruskal-Wallis nonparametric tests. RESULTS: Bronchial cross-sectional areas and lung attenuation did not vary significantly compared with baseline values following bronchial challenge in healthy volunteers or patients with asthma. However, in patients with asthma, bronchial cross-sectional areas were significantly smaller than in healthy volunteers, except after inhalation of salbutamol. Lung attenuation and anteroposterior attenuation gradient were significantly higher in patients with asthma than in healthy patients (P <.001). Air-trapping scores were significantly higher after methacholine challenge. CONCLUSION: Helical thin-collimation CT at controlled lung volume and at full expiration associated with bronchial challenge may help evaluate bronchoreactivity and inflammation in mild intermittent asthma. PMID- 11930066 TI - Lymphangioleiomyomatosis: correlation of qualitative and quantitative thin section CT with pulmonary function tests and assessment of dependence on pleurodesis. AB - PURPOSE: To explore the relationship between findings at thin-section computed tomography (CT) and pulmonary function tests in lymphangioleiomyomatosis (LAM) and to evaluate the influence of pleurodesis on this relation and the effectiveness of quantitative versus qualitative CT in the assessment of disease severity. MATERIALS AND METHODS: Thirty-seven patients with LAM (17 with pleurodesis) underwent CT and pulmonary function tests. The severity of pulmonary cystic involvement was graded qualitatively by two independent readers and measured quantitatively at CT with a thresholding technique. Relationships between findings at CT and pulmonary function tests and the influence of pleurodesis on these findings were assessed with regression analysis and analysis of covariance. RESULTS: Qualitative ratings had good agreement between observers (kappa = 0.75). Quantitative CT had good repeatability and showed significant correlation with the percent predicted forced expiratory volume in 1 second (FEV(1)%) (r = 0.67, P <.001), percent predicted diffusing capacity of lung for carbon monoxide (DLCO%) (r = 0.48, P <.005), percent predicted ratio of residual volume to total lung capacity (RV/TLC%) (r = -0.65, P <.001), and percent predicted TLC (r = 0.34, P <.04). Quantitative CT results were somewhat better than qualitative CT results. The standard error of the FEV(1)% for the quantitative CT was about 85% of that for the qualitative CT. Pleurodesis had no statistically significant effect on the slope of the regression line between quantitative CT findings, FEV(1)%, and DLCO% (corrected for alveolar volume). The slope between quantitative CT and RV/TLC% was significantly (P =.044) more negative in patients with pleurodesis. CONCLUSION: Qualitative and quantitative CT findings correlate with pulmonary dysfunction over a wide range of disease severity in patients with LAM. Pleurodesis influences the relationship between CT measurements and pulmonary function test results. PMID- 11930067 TI - Detection of lung nodules on digital chest radiographs: potential usefulness of a new contralateral subtraction technique. AB - PURPOSE: To evaluate the potential usefulness of a contralateral subtraction technique developed for radiologists' performance in the detection of subtle lung nodules on chest radiographs. MATERIALS AND METHODS: Fifty chest radiographs (25 normal and 25 abnormal with a subtle lung nodule) that were digitized with a 0.175-mm pixel size and 4,096 gray levels were used. Twelve radiologists (10 attending and two residents) participated in observer tests and read both original and contralateral subtraction images with a sequential testing method. Radiologists' performance was evaluated by means of receiver operating characteristic analysis with use of a continuous rating scale. The beneficial and detrimental effects of the contralateral subtraction technique on the radiologists' performance were also evaluated. RESULTS: The area under the receiver operating characteristic curve values obtained without and with contralateral subtraction images were 0.926 and 0.962, respectively. Results indicated that the contralateral subtraction images significantly (P <.05) improved diagnostic accuracy, particularly for radiologists with limited experience. CONCLUSION: The contralateral subtraction technique can assist radiologists in the correct identification of subtle lung nodules on chest radiographs. PMID- 11930068 TI - Exercise MR imaging in the assessment of pulmonary regurgitation and biventricular function in patients after tetralogy of fallot repair. AB - PURPOSE: To assess the responses of pulmonary regurgitation (PR) and biventricular function to submaximal exercise by using a magnetic resonance (MR) imaging exercise protocol with young adult patients who underwent tetralogy of Fallot repair at a young age. MATERIALS AND METHODS: Fifteen patients with corrected tetralogy of Fallot (mean age, 17.5 years +/- 2.5 [SD]) underwent MR imaging at rest and during exercise for the evaluation of PR and biventricular function. Results were compared with findings from 16 control subjects (mean age, 17.5 years +/- 2.3). Mean age at tetralogy of Fallot repair was 2.1 years +/- 1.6, and mean follow-up time after repair was 15.4 years +/- 2.6. Exercise level at MR imaging was calculated individually and corresponded to 60% of peak oxygen uptake. The parameters of cardiac function obtained at rest and during exercise were compared by using a paired t test. An unpaired t test was used to compare parameters of cardiac function between patients and control subjects. RESULTS: PR decreased during exercise (from 27 mL/m(2) +/- 17 to 23 mL/m(2) +/- 15; P =.012). At rest, right ventricular (RV) ejection fraction was normal (>47%) in 80% of patients. RV response to exercise in the patient group was abnormal compared with response in the control group, as demonstrated by an increase in RV end-diastolic volume index (132 mL/m(2) +/- 36 to 137 mL/m(2) +/- 38; P =.041) and no significant change in end-systolic volume index or ejection fraction. In only one patient, RV ejection fraction increased by more than 5%. Left ventricular response was not different between patients and control subjects. CONCLUSION: MR imaging is well suited to assess cardiac response to exercise, and findings revealed a decrease in PR and an abnormal RV response to exercise in patients with corrected tetralogy of Fallot. PMID- 11930069 TI - Techniques for the detection of coronary atherosclerosis: multi-detector row CT coronary angiography. AB - PURPOSE: To investigate the accuracy of different computed tomographic (CT) reformation techniques in assessing the coronary arteries. MATERIALS AND METHODS: Sixty-four patients undergoing both multi-detector row CT and invasive coronary angiography were consecutively included in a retrospective study. CT scans were obtained with collimation of 4 x 1 mm, pitch of 1.5, and rotation time of 500 msec. Retrospective electrocardiographic gating was used for image reconstruction, with 1.25-mm section thickness and 0.5-mm increment. The CT data set of each patient was evaluated by independent observers using transverse scanning, virtual endoscopic, and three-dimensional reformation and multiplanar reformation. RESULTS: Hemodynamically relevant stenoses (>50%) were detected with highest sensitivity at transverse scanning (58 of 79 [73.4%] stenoses), followed by virtual endoscopic (38 of 79 [48.1%] stenoses) and three-dimensional reformation (34 of 79 [43.0%] stenoses), and multiplanar reformation (37 of 79 [46.8%] stenoses). Atherosclerotic plaques were identified with comparable sensitivities at transverse scanning (143 of 218 plaques [65.6%]) and at three dimensional (139 of 218 [63.8%] plaques) and virtual endoscopic reformation (136 of 218 [62.4%] plaques). Multiplanar reformation had distinctly poorer results (217 of 218 [58.3%] plaques). Combined interpretation with all four techniques increased sensitivity to 74.7% (59 of 79) for stenosis and 71.6% (156 of 218) for atherosclerosis. Calculated overall specificity was 91.4% or greater. Sufficient vascular evaluation was possible only in vessels larger than 1.6 mm in diameter. Thus, even in patients with heart rates below 60 bpm, only 80.0% of all coronary segments could be visualized, while at higher frequencies, visibility decreased to 66.2%. CONCLUSION: Although multi-detector row CT is a favorable alternative procedure in evaluating coronary arteries, its clinical value still is restricted to low heart rates and proximal coronary arterial segments. PMID- 11930070 TI - Malignant lesions initially subjected to short-term mammographic follow-up. AB - PURPOSE: To determine whether systematically evaluated criteria for probably benign lesions were actually applied to lesions placed into that category. MATERIALS AND METHODS: A search of the mammography database yielded 295 cases that were initially followed up with short-term interval mammography but eventually received a biopsy recommendation for the same breast. Of the 83 malignancies (81 patients) for which mammograms and pathology reports were available for review, 51 malignancies corresponded to the lesions for which short term follow-up was recommended. Each case was retrospectively reviewed to determine whether the lesion followed up represented the subsequently diagnosed malignancy. Each lesion was characterized with appropriate Breast Imaging Reporting and Data System descriptors, based on the mammographic imaging available when short-term follow-up was first recommended. These characteristics were then used to determine if, in retrospect, the mammographic appearance met previously published criteria for probably benign lesions. RESULTS: Of the 51 malignancies, 23 (45%) appeared mammographically as microcalcifications, 12 (24%) as masses, four (8%) as architectural distortion, and 12 (24%) as developing densities. None fulfilled strict criteria for a probably benign lesion when reviewed in retrospect. Forty-seven (92%) of 51 lesions had already demonstrated progression at the time of follow-up recommendation. CONCLUSION: Short-term mammographic follow-up is often recommended for lesions that, in retrospect, do not fulfill established diagnostic criteria for probably benign lesions. PMID- 11930071 TI - Breast US: assessment of technical quality and image interpretation. AB - PURPOSE: To determine whether ultrasonography (US) of the breast performed at a wide range of clinical practices conforms to the American College of Radiology (ACR) standards for quality and to assess the interpretations of breast sonograms. MATERIALS AND METHODS: Static images from 152 breast US examinations performed at 86 institutions were evaluated for compliance with ACR guidelines for breast US hardware, technical factors, imaging protocol, and image annotation. Official interpretations submitted by the referring facilities were compared with static images submitted by the facility. Discrepancies were confirmed by two dedicated breast radiologists after repeat imaging, short interval follow-up imaging, or biopsy. RESULTS: A total of 60.5% of cases did not comply with at least one ACR guideline on breast US and included 9.2% of cases with inadequate equipment, 14.7% of cases with inappropriate focal zone placement, at least 14% of cases with static images in only one imaging plane, and 25% of cases with incomplete patient identifiers. Clinically relevant interpretation errors and interpretation discrepancies were confirmed in 23 (15.1%) of 152 cases. CONCLUSION: The majority of breast US examinations did not comply with at least some of the standards for quality set forth by the ACR. Attention to these basic standards could substantially improve image quality. PMID- 11930072 TI - The pseudocalculus sign. PMID- 11930073 TI - Evaluation of patients with jejunostomy tubes: imaging findings. AB - PURPOSE: To determine the frequency and nature of abnormalities observed on radiographs after placement of jejunostomy (J) tubes for enteral nutrition. MATERIALS AND METHODS: Radiology database review revealed that 280 studies of the J tube or of the small bowel with water-soluble contrast material and/or barium sulfate were performed in patients during 10 years. Review of the radiologic reports revealed abnormalities related to the placement of tubes in 105 (38%) cases. Images were reviewed to determine abnormalities in these 105 cases. Radiologic, medical, and surgical records were also reviewed to determine the clinical course and any subsequent interventions. RESULTS: One or more complications were detected in 40 (14%) of 280 cases: small-bowel obstruction in 17 (6%) cases, nonobstructive small-bowel narrowing in six (2%), extraluminal tracks or collections in seven (2%), extravasation of contrast material to the skin in 11 (4%), jejunal hematomas in five (2%), and intussusceptions in four (1%). Mechanical problems related to the tube were detected in 52 (19%) cases, including coiling, kinking, or knotting of the tube in 38 (14%), malpositioning in five (2%), retrograde flow in four (1%), occlusion in four (1%), and a hole in one (<1%). Focal thickening of small-bowel folds was detected in 24 (9%) cases. CONCLUSION: Radiographs in 280 patients with J tubes revealed one or more complications that resulted from tube placement (40 [14%] cases), mechanical problems related to location or function of the tube (52 [19%] cases), and development of focally thickened small-bowel folds (24 [9%] cases). PMID- 11930076 TI - MR Imaging of the heart with cine true fast imaging with steady-state precession: influence of spatial and temporal resolutions on left ventricular functional parameters. AB - The influence of changes in spatial and temporal resolutions on functional parameters in the left ventricle (LV) were investigated with magnetic resonance (MR) imaging with a modified true fast imaging with steady-state precession, or FISP, two-dimensional sequence that provided temporal resolution of 21-90 msec and spatial resolution of 1-3 mm. MR imaging in the heart was performed in 15 healthy volunteers. A decrease in LV functional parameters was observed with reduced spatial and temporal resolutions. The influence of temporal resolution was more relevant. PMID- 11930075 TI - Hepatic tumors treated with percutaneous radio-frequency ablation: CT and MR imaging follow-up. AB - PURPOSE: To describe the appearance of hepatic tumors treated with radio frequency (RF) ablation on computed tomographic (CT) and magnetic resonance (MR) images and the pattern of residual tumor at the site of RF ablation and to assess prospectively the sensitivity, specificity, and positive and negative predictive CT and MR imaging values in the evaluation of RF treatment. MATERIALS AND METHODS: Thirty-one patients with 50 tumors (nine hepatocellular carcinomas and 41 metastases) treated with RF ablation underwent CT and MR imaging on the same day at 2, 4, and 6 months; CT was performed every 3 months thereafter. CT and MR findings were interpreted separately and prospectively by two reviewers with consensus. For both imaging techniques, appearance of the treated area, treatment efficacy, and complications were assessed at each time. Sensitivity and specificity were determined by using the McNemar test. RESULTS: After a mean follow-up of 19 months, nine tumors showed local regrowth. At 2 months, MR imaging depicted more local regrowths (eight of nine; sensitivity, 89%) than did CT (four of nine; sensitivity, 44%) but without significant differences (P =.12). In two cases, only T2-weighted imaging depicted local regrowth. All nine lesions became conspicuous at 4-month follow-up with both techniques. At 2 months, thin peripheral rim enhancement and arterioportal shunting were found in 24% and 12%, respectively, of the treated tumors. These findings disappeared thereafter and are not linked to tumor regrowth. CONCLUSION: Despite the small number of patients, CT and MR imaging may depicted all local regrowth at 4 months or sooner. MR imaging may have an edge over CT in the early detection of local regrowth. PMID- 11930074 TI - MR colonography with barium-based fecal tagging: initial clinical experience. AB - PURPOSE: To assess a strategy for fecal tagging with barium sulfate as an inexpensive tagging agent in conjunction with magnetic resonance (MR) colonography in patients suspected of having colorectal lesions. MATERIALS AND METHODS: Twenty-four patients suspected of having colonic lesions because of rectal bleeding, positive fecal occult blood test results, or altered bowel habits underwent MR colonography and subsequent conventional colonoscopy. A 200 mL dose of a barium sulfate-containing contrast agent was ingested with each of four low-fiber meals, beginning 36 hours before the examination. For MR colonography, the colon was filled with tap water. Gadobenate dimeglumine was injected intravenously. Images were acquired 75 seconds after gadobenate dimeglumine administration by using only a T1-weighted three-dimensional gradient echo sequence. Images were reviewed by two radiologists blinded to conventional colonoscopic data. By using colonoscopy as the reference standard, sensitivity and specificity of MR colonography were determined for detecting colorectal masses. RESULTS: On the basis of MR colonography, 15 polyps of 5-20 mm and 10 carcinomas were detected and later confirmed with conventional colonoscopy. Conventional colonoscopy depicted three additional lesions less than 8 mm in diameter. Thus, sensitivity of MR colonography was 89.3% (25 of 28) for lesions and 91.7% (22 of 24) for patients. CONCLUSION: Barium-tagged MR colonography obviates bowel cleansing and depicts all lesions exceeding 8 mm in diameter. PMID- 11930077 TI - Thoracic aortic dissection and aneurysm: evaluation with nonenhanced true FISP MR angiography in less than 4 minutes. AB - Comprehensive aortic magnetic resonance (MR) examinations currently include multiple nonenhanced and contrast material-enhanced sequences. The authors hypothesized that the nonenhanced true fast imaging with steady-state precession (FISP) portion alone of their comprehensive imaging protocol would be adequate to confidently confirm or exclude dissection or aneurysm of the aorta. In a retrospective review of 29 comprehensive thoracic aortic MR examinations, nonenhanced true FISP MR imaging alone was 100% accurate for determining the presence or absence of dissection or aneurysm. PMID- 11930078 TI - Automated generation of curved planar reformations from volume data: method and evaluation. AB - The authors developed and evaluated a method to automatically create interactive vascular curved planar reformations with computed tomographic (CT) angiographic data. The method decreased user interaction time by 86%, from 15 to 2 minutes. Expert reviewers were asked to indicate their confidence in differentiating automatically created images from clinical-quality manually produced images. The area under the receiver operating characteristic curve was 0.45 (95% CI: 0.39, 0.51), and a test of equivalency indicated that reviewers could not distinguish between images. They also graded image quality as equivalent to that with manual methods and found fewer artifacts on automatically created images. Automatic methods rapidly produce curved planar reformations of equivalent quality with reduced time and effort. PMID- 11930079 TI - Potential of in vivo proton MR spectroscopy in the assessment of breast lesions without the use of contrast agent. PMID- 11930080 TI - Accuracy of gray-scale and color Doppler US and serum markers as predictors of prostate carcinoma. PMID- 11930084 TI - The message and the messenger: delivering RNA in neurons. AB - Synaptic plasticity results in enduring changes in synaptic function. Localized protein synthesis is part of this process. Kosik and Krichevsky describe how a dynamic macromolecular structure, the RNA granule, may be a key element contributing to changes in protein production leading to synaptic plasticity. PMID- 11930086 TI - Micronutrient supplementation and infection: a double-edged sword? PMID- 11930085 TI - Iron metabolism and requirements in early childhood: do we know enough?: a commentary by the ESPGHAN Committee on Nutrition. PMID- 11930087 TI - Nutritional interventions in pediatric HIV: it's hard to hit a moving target. PMID- 11930088 TI - Constipation: is there a new approach? PMID- 11930089 TI - Post-transplant lymphoproliferative disorders: making sense of the proliferation of potential therapies. PMID- 11930090 TI - Preparing your curriculum vitae. PMID- 11930091 TI - Effects of nutritional rehabilitation on intestinal function and on CD4 cell number in children with HIV. AB - BACKGROUND: A complex interplay of malnutrition, intestinal dysfunction, and immune impairment increases the progression of human immunodeficiency virus (HIV) disease in children. The authors tested the hypothesis that nutritional support improves intestinal and immune functions in children infected with human immunodeficiency virus (HIV). METHODS: A questionnaire was circulated through reference centers for pediatric HIV infection to evaluate the effects of nutritional rehabilitation, total parenteral nutrition (TPN) and enteral nutrition (EN), in children. Information included changes in body weight, CD4 cell numbers, and intestinal absorption-as judged by the xylose load-before and after clinical nutritional support and the outcome of children. RESULTS: Sixty two children underwent nutritional support: 46 received TPN and 16 received EN. All but three had full-blown acquired immunodeficiency syndrome, and all were severely malnourished. Baseline clinical conditions were worse in children receiving TPN than in those receiving EN. Intestinal dysfunction was detected in all children who received xylose oral load. A significant increase in CD4 cell count, xylose levels, and body weight followed EN. A similar pattern was observed after TPN, but none of the parameters significantly changed. Twenty-seven children who received TPN and three who received EN eventually died. Fourteen who received TPN and eight who received EN were shifted to oral feeding, and five who received TPN and five who received EN continued with clinical nutritional support at the end of the observation period. CONCLUSIONS: Nutritional intervention may restore intestinal absorption and increase CD4 cell numbers. The efficacy of nutritional intervention is enhanced if provided before a terminal stage of HIV infection. These data provide evidence of a close association among nutritional condition, intestinal absorption, and immune impairment. PMID- 11930092 TI - Polyethylene glycol without electrolytes for children with constipation and encopresis. AB - BACKGROUND: Children with functional constipation and encopresis benefit from behavior modification and from long-term laxative medication. Polyethylene glycol without electrolytes has become the first option for many pediatric gastroenterologists. METHODS: Twenty-eight children treated with polyethylene glycol without electrolytes were compared with 21 children treated with milk of magnesia to evaluate the efficiency, acceptability, side effects, and treatment dosage of polyethylene glycol in long-term treatment of functional constipation and encopresis. Children were rated as "doing well," "improved," or "not doing well," depending on resolution of constipation and encopresis. RESULTS: At the 1 , 3-, 6-, and 12-month follow-ups, bowel movement frequency increased and soiling frequency decreased significantly in both groups. At the 1-month follow-up, children on polyethylene glycol were soiling more frequently (P < 0.01) and fewer were improved (P < 0.01). At the 3- and 6-month follow-ups, both groups had similarly improved. At the 12-month visit, 61% of children on polyethylene glycol and 67% of children on milk of magnesia were doing well. Children on polyethylene glycol soiled more frequently (P < 0.01). None refused polyethylene glycol, but 33% refused to take milk of magnesia. The mean initial treatment dosage of polyethylene glycol was 0.6 +/- 0.2 g/kg daily. Polyethylene glycol had no taste, and no loss of efficacy occurred. Polyethylene glycol did not cause clinically significant side effects. CONCLUSIONS: Polyethylene glycol without electrolytes is an alternative for long-term management of children with constipation and encopresis. PMID- 11930093 TI - Treatment of childhood encopresis: a randomized trial comparing three treatment protocols. AB - OBJECTIVES: To compare short- and long-term effectiveness of three additive treatment protocols in children experiencing chronic encopresis. METHODS: Children, 6 to 15 years of age, who experienced at least weekly fecal soiling for 6 months or longer were eligible for the study. Children were randomly assigned to a group that received intensive medical therapy (IMT), a group that received intensive medical therapy plus a behavior management program called enhanced toilet training (ETT), or a group that received intensive medical therapy with enhanced toilet training and external anal sphincter electromyographic biofeedback (BF). Data concerning toileting habits were collected for 14 consecutive days before an initial visit, and at 3, 6, and 12 months after initiation of therapy. All data were collected using a computerized voice-mail system that telephoned the families each day. At 12 months, children were classified as significantly improved (reduction in soiling, P < 0.001) or cured ( 0.90, P < 0.001 in each case). Response to treatment during the first 2 weeks of therapy was highly predictive of outcome at 3, 6, and 12 months (P < 0.0001). Children in the ETT group used less laxative medication (P < 0.04) and required fewer treatment contacts (P = 0.08) than children in the IMM group. All three treatments resulted in significant increases in daily bowel movements passed in the toilet and self-initiated toileting, and resulted in decreases in average daily soiling at 3, 6, and 12 months (P < 0.05). CONCLUSIONS: Enhanced toilet training is somewhat more effective in treating childhood encopresis than either intensive medical therapy or anal sphincter biofeedback therapy. Although similar total cure rates at 1 year can be expected with these three forms of therapy, enhanced toilet training results in statistically significant decreases in the daily frequency of soiling for the greatest number of children. PMID- 11930094 TI - Early constipation and toilet training in children with encopresis. AB - OBJECTIVE: To evaluate the frequency of predisposing factors for encopresis before and during toilet training, comparing children with primary and secondary encopresis. METHODS: In this retrospective study, questionnaires from the initial evaluation at an encopresis clinic at a tertiary care pediatric hospital were reviewed for the presence or absence of factors in the first 2 years of life, for toilet training practices, and for disruptive events during the training process. Children younger than 48 months or those with organic defecation disorders were excluded. RESULTS: In 411 children with encopresis, the reported frequency of predisposing factors included constipation in 35%, and previous treatment for constipation in 24%. Toilet training was initiated before age 2 years in 26% and after age 3 years in 14%. Interruption of toilet training and punishment were seen more in primary encopresis than in secondary encopresis (50% versus 23%; P < 0.05) and (52% versus 26%; P < 0.05) respectively. Constipation (30% versus 18%; P < 0.05) and abdominal pain (23% versus 9%; P <0.0:5) during toilet training were more common in primary encopresis as was fear of the toilet (47% versus 10%; P < 0.05). CONCLUSIONS: In children with encopresis, early difficult defecation, previous treatment for constipation, and early initiation of toilet training were less common than expected. Children with primary encopresis did not have an increased incidence of early constipation or invasive treatments compared with those with secondary encopresis. However, children with primary encopresis did have more difficult and disruptive toilet training experiences. PMID- 11930095 TI - Anti-CD20 monoclonal antibody (Rituximab) treatment for Epstein-Barr virus associated, B-cell lymphoproliferative disease in pediatric liver transplant recipients. AB - OBJECTIVES: Anti-B-cell immunotherapy has been used with success in adults with posttransplant B-cell lymphoproliferative disease (PTLD), but such treatment has rarely been reported in children. We report the outcome of anti-CD20 antibody (rituximab) therapy for Epstein-Barr virus (EBV)-associated PTLD in six pediatric liver transplant recipients. METHODS: In these six patients, PTLD was diagnosed within 2 to 4 months after transplantation. The tumors were classified as monomorphic or polymorphic B-cell infiltrate expressing CD20 antigen and EBV genome. Anti-CD20 therapy was associated with withdrawal of tacrolimus or ciclosporine therapy in all patients. Intravenous rituximab was administered at 375 mg/m2 once a week for 3 to 4 consecutive weeks. RESULTS: Rituximab treatment was associated with decreased EBV load, disappearance of abnormal serum immunoglobulin concentration, and disappearance of tumoral masses, which occurred 1 to 2.5 months after treatment onset. Despite rituximab therapy, one patient was diagnosed subsequently with a cerebral tumor. Five patients experienced acute liver graft rejection episodes within 10 days to 2.5 months after beginning treatment. In these patients, immunosuppression was reintroduced, but three children experienced fatal chronic rejection, whereas two experienced complete tumor remission. Three children are alive and in complete remission, with normal liver tests, 15 months to 3 years after PTLD onset. CONCLUSIONS: Rituximab therapy is an interesting approach for children with early EBV-associated PTLD after liver transplantation. It does not prevent cerebral localization, and rapid resumption of immunosuppression may be advisable to prevent lethal chronic liver graft rejection. PMID- 11930096 TI - Effect of human milk and epidermal growth factor on growth of human intestinal Caco-2 cells. AB - BACKGROUND: Epidermal growth factor (EGF) in human milk has been thought to be mitogenic for cell growth. This study investigated the effects of human milk and EGF on the growth of human intestinal Caco-2 cells to determine whether the action occurred through regulation of the cell cycle or through c-jun expression. METHODS: Cells were incubated with 5% human milk, 0.375 nmol/L EGF (relevant to EGF concentration in 5% human milk, 0.05 x EGF), 7.5 nmol/L EGF (1 x EGF), or 75 nmol/L EGF (10 x EGF). Cell numbers; cellular RNA, DNA, and protein concentrations; DNA content in the cell cycle, and expressions of c-Jun protein and mRNA were analyzed. RESULTS: Cell numbers increased in the 1 x and 10 x EGF groups at 48 hours. Cellular RNA increased in the 5% human milk and 10 x EGF groups. DNA and protein contents increased in the 1 x and 10 x EGF groups. The 1 x and 10 x EGF groups increased DNA content in the G1 phase compared with the 5% human milk group at 24 hours. The greatest c-jun protein expression was 2.6, 1.4, 1.8, and 1.9 times the control, and the c-jun mRNA increased by 202%, 14%, 150%, and 181%, respectively, in the 5% human milk, 0.05 x, 1 x, and 10 x EGF groups. CONCLUSIONS: In a dose-dependent manner, EGF stimulated intestinal growth in vitro, by increasing DNA content in the G1 phase and c-jun mRNA expression. However, low concentrations of human milk (5%) and its equivalent EGF did not affect cell growth. PMID- 11930097 TI - Management of intractable constipation with antegrade enemas in neurologically intact children. AB - OBJECTIVES: To assess the benefit of antegrade enemas in children with severe constipation who were referred to a tertiary care center. METHODS: From 1997 to 1999, 12 children (9 male, aged 8.7 +/- 4.4 years) underwent cecostomy placement. All children were neurologically normal and had been extensively examined to rule out organic causes of constipation. Follow-up included a questionnaire to interview caregivers 13.1 +/- 8.5 months after cecostomy placement. RESULTS: For all children, antegrade enemas led to improvement in the number of bowel movements / week (7.1 versus 1.4, P < 0.005), number of soiling accidents / week (1.0 versus 4.7, P < 0.01), abdominal pain score (0.9 versus 2.9, P < 0.005), emotional health score (3.6 versus 1.9, P < 0.005), overall health score (3.6 versus 1.7, P < 0.005), number of medications used for constipation (0.8 versus 4.0, P < 0.005), number of missed school days / month (1.5 versus 7.5, P < 0.02), and number of physician office visits / year (9.2 versus 24.0, P < 0.05). Irrigation solutions used for the antegrade enemas included polyethylene glycol (67%), saline and glycerin solution (25%), and phosphate enema (8%), administered everyday in seven children and every other day in five children. Adverse events included skin breakdown and granulation tissue in one patient, leakage of irrigation solution in one patient, and dislodging of the tube in two patients. Five patients discontinued the use of antegrade enemas within a mean of 14.6 +/- 9.1 months after beginning treatment. CONCLUSION: Antegrade enemas through a cecostomy are a safe and satisfactory option for children who are neurologically intact and who have severe constipation that does not respond to medical treatment. PMID- 11930098 TI - Gastrointestinal malformations, associated congenital abnormalities, and intrauterine growth. AB - BACKGROUND: In contrast with other malformations, congenital anomalies of the gastrointestinal tract have been scarcely investigated. METHODS: The prevalence of gastrointestinal malformations with special reference to associated disorders and intrauterine growth was retrospectively analyzed in the newborn infants admitted to the Neonatal Intensive Care Unit of the Department of Pediatrics, University of Pecs, Hungary, in the 14-year period between 1987 and 2000. RESULTS: Of 4,241 neonates with gastrointestinal malformations, 241 (5.68%) had a total of 304 malformations (excluding Hirschsprung disease). In 133 patients, the gastrointestinal anomalies were observed as one of multiple malformations; a specific syndrome or association was diagnosed in 36 cases. Skeletal disorders were the most frequently associated anomalies. Intrauterine growth retardation was found in a large number of patients with both isolated and multiple gastrointestinal malformations (38.9% and 30.8%, respectively). CONCLUSIONS: Gastrointestinal malformations often are complicated by skeletal anomalies and intrauterine growth retardation. The association among these disorders requires further investigation. However, from a practical point of view, this association should be considered in treating affected patients. PMID- 11930099 TI - Listeria meningitis after treatment with infliximab. PMID- 11930100 TI - Inferior vena cava occlusion and protein-losing enteropathy after liver transplantation in children. PMID- 11930101 TI - Cimetropium bromide in the treatment of crisis in infantile colic. AB - BACKGROUND: Treatment of infantile colic remains an open issue. In Italy, cimetropium bromide is used extensively to treat infantile colic. The aim of this randomized, double-blind, placebo-controlled clinical trial was to investigate the effectiveness and side effects of cimetropium bromide in the treatment of infants with colic crisis. METHODS: Ninety-seven infants with colic were enrolled. The diagnosis of infantile colic in healthy infants with regular growth, aged 15 to 60 days was made according to the criteria of Wessel. The infants were divided into two groups, one treated with cimetropium bromide (1.2 mg/kg) and the other treated with placebo at onset of each crisis for 3 days. Duration of crying and side effects were recorded daily in a structured diary for the 3 days of therapy. Statistical analysis was performed using the chi-square and t tests. RESULTS: Eighty-six infants completed the trial. The average duration of crying for each crisis was 17.3 +/- 12.6 minutes in the cimetropium bromide group and 47.5 +/- 28.5 minutes in the placebo group (P < 0.005). Response to cimetropium bromide was 74%. Response to placebo was 33% (P < 0.05). Side effects did not differ significantly between the two groups, except sleepiness, which increased in the infants treated with cimetropium bromide. CONCLUSION: Cimetropium bromide was more effective than placebo in reducing the duration of crying in children with infantile colic. The use of the anticholinergic drugs, aside from the dicyclomine because of its dangerous side effects, should be revaluated for treating infantile colic. PMID- 11930102 TI - Successful treatment of metastatic Crohn disease with infliximab. PMID- 11930103 TI - Celiac disease screening of Turkish children with recurrent abdominal pain. PMID- 11930104 TI - Effect of growth hormone therapy in patients with Crohn disease. PMID- 11930105 TI - Effects of several flavonoids on the growth of B16F10 and SK-MEL-1 melanoma cell lines: relationship between structure and activity. AB - Although flavonoids seem to be capable of acting at all stages of the carcinogenic process, little information is available on their action in melanoma cell lines. The aim of this study was to assess the response of B16F10 and SK-MEL 1 melanoma cell lines to treatment with six different flavonoids after 24 and 72 h of exposure and to relate the response to their structure. We then compared the findings with those for melphalan treatment. When cultures were treated for 24 h, only slight inhibition at the highest concentrations (25 and 50 microM) of tangeretin and luteolin were observed, whereas melphalan caused a dose-related inhibition of growth at all concentrations. Quercetin, hesperetin, 7,3' dimethylhesperetin and eriodictyol did not produce any effect at 24 h on B16F10 or SK-MEL-1 cells, results which point to the low toxicity of flavonoids. After 72 h of exposure culture growth was inhibited by 7,3'-dimethylhesperetin at 50 microM, but lower concentrations had no effect. Tangeretin was the most effective of the flavonoids in inhibiting B16F10 and SK-MEL-1 cell growth, showing a clear dose-response curve after 72 h. These results suggest that the absence of the C2 C3 double bond on hydroxylated flavonoids results in a loss of effect on both the cell lines, while the higher activity of tangeretin compared with 7,3' dimethylhesperetin suggests that the presence of at least three adjacent methoxyl groups confers a more potent antiproliferative effect. PMID- 11930106 TI - What is the relationship between P-glycoprotein and adhesion molecule expression in melanoma cells? AB - A number of studies have reported that increased P-glycoprotein expression in drug-resistant tumour cells may be associated with decreased expression of a family of surface glycoproteins. However, despite its potential biological and clinical relevance, this phenomenon has not been extensively studied. In this study the phenotypic alterations that are associated with the acquisition of the multidrug-resistant phenotype in tumour cells, together with drug transporter overexpression, were investigated in human melanoma cells. The expression of cell adhesion molecules was analysed in a panel of multidrug-resistant melanoma cell lines (M14Dx) showing different degrees of resistance to doxorubicin and different levels of the expression of the drug transporter P-glycoprotein. In particular, expression of intercellular adhesion molecule-1 (ICAM-1), CD44, very late activation antigen (VLA)-5 and VLA-2 was determined by flow cytometry in the different resistant cell lines. A progressive downregulation of all the adhesion molecules examined was revealed in M14Dx cells, in parallel with an increasing level of expression of the drug transporter P-glycoprotein. The results obtained raise the question of the role of P-glycoprotein in the invasive and metastatic behaviour of tumour cells. PMID- 11930107 TI - Melanin affinity of N-(2-diethylaminoethyl)-4-iodobenzamide, an effective melanoma imaging agent. AB - The cellular uptake and incorporation in macromolecules of iodine-125 labelled N (2-diethylaminoethyl)-4-iodobenzamide ([125I]BZA), a melanoma imaging agent, was studied using human melanoma cells M3Dau (amelanotic) and M4Beu (melanotic). The interaction between [125I]BZA and synthetic melanin was examined in various conditions of incubation. The results showed that uptake was high only for M4Beu, whereas the incorporation in trichloroacetic acid-precipitable proteins was very low for both model cell lines, with no correlation with melanin content. Experiments with synthetic melanin showed that BZA binding to melanin was saturable and reversible, and involved several types of interaction. The influence of the ionic environment indicated that electrostatic forces play a role in the affinity, and the decrease in binding produced by the presence of an alcohol in the medium suggested that hydrophobic interactions may be involved in the binding mechanism. This was supported by the Scatchard analysis, which revealed two classes of binding sites, and the determination of two association constants (K1 = 3.9 +/- 1.9 x 106/M and K2 = 2.9 +/- 0.9 x 104/M). The affinity of BZA for melanin might explain the good results obtained in a phase II clinical trial for the diagnosis of malignant melanoma metastases, in which the specificity was 100%. PMID- 11930108 TI - Priming effects of GM-CSF, IFN-gamma and TNF-alpha on human neutrophil inflammatory cytokine production. AB - Identifying and evaluating the priming agents for cytokine release by neutrophils might be helpful in controlling the innate immune response of the host. In the present study we examined the role of granulocyte/macrophage colony-stimulating factor (GM-CSF), interferon-gamma (IFN-gamma) and tumour necrosis factor-alpha (TNF-alpha) as priming agents for interleukin (IL)-1beta, IL-6 and TNF-alpha production by stimulated neutrophils from control subjects and malignant melanoma patients. When the cells from controls and patients were preincubated with primer agents, opsonized zymosan-stimulated inflammatory cytokine production was enhanced. The major neutrophil-priming factor for IL-6 secretion by polymorphonuclear leukocytes (PMNs) in the control and patient groups was TNF alpha. However, GM-CSF and IFN-gamma are also significant primers. GM-CSF priming was critical for the release of TNF-alpha from PMNs in control and melanoma patients. The ability of GM-CSF, IFN-gamma and TNF-alpha to serve as effective priming agents for inflammatory mediator production by PMNs revealed a new role for these cytokines in the innate immune response of the melanoma-bearing host. PMID- 11930109 TI - CD80-mediated induction of immunostimulation in two ocular melanoma cell lines is augmented by interferon-gamma. AB - Although the transfection of the T-cell costimulatory molecule CD80 cDNA into human tumours can augment their immunogenicity in vitro, its expression alone is ineffective in many tumour systems. We evaluated the influence of CD80 expression on the immunostimulatory activity of ocular melanoma cell lines and determined whether IFN-gamma could enhance the effect. Two ocular melanoma cell lines were transfected with CD80 cDNA. The immunostimulatory capacity of the CD80+ transfectants was determined by their ability to stimulate the proliferation of allogeneic peripheral blood mononuclear cells (PBMC). The influence of additional accessory molecules on PBMC proliferation was assessed by pre-treating the CD80 transfectants with IFN-gamma. The CD80+ transfectants induced proliferation of allogeneic PBMC. IFN-gamma treatment of the tumour cells induced upregulated expression of MHC class I, de novo expression of MHC class II and CD54, and enhanced the ability of the CD80+ transfectants to stimulate PBMC proliferation. CD4+ T cells were not required for the proliferative response against untreated CD80+ tumour cells but were necessary for the augmentation of proliferation observed following IFN-gamma treatment. CD80+ ocular melanoma cells possess immunostimulatory potential which is augmented by IFN-gamma induced upregulation of cell surface molecules. Further studies on the role of costimulatory molecules in inducing anti-tumour immunity in ocular melanoma may help to define new strategies for application of immunotherapeutic approaches to treat this aggressive disease. PMID- 11930110 TI - Overexpression of HER-2 in thick melanoma. AB - In this study we evaluated the overexpression status of HER-2 and its prognostic significance on survival in patients with thick cutaneous malignant melanoma. The immuno-alkaline phosphatase antigen detection technique was applied to archival diagnostic material from 51 patients with primary lesions measuring >or= 10 mm in Breslow thickness. Eleven additional patients with primary lesions measuring or = 4 h on continued exposure. Ten minutes pretreatment with 1 or 5 microM tetramethylpyrazine dose dependently and significantly attenuated the kainate-induced damage. Taken together, the results suggest that multiple mechanisms including protection of mitochondria, decrease in free radical generation and scavenging of free radicals might be involved in TMP's protection against kainate induced cell toxicity. PMID- 11930175 TI - Autocorrelation analyses of magnetoencephalographic alpha waves in relation to subjective preference for a flickering light. AB - Human cortical responses corresponding to the subjective preference for a flickering light of varying period were investigated. Paired-comparison tests were performed to examine the subjective preference for a flickering light, and MEG was recorded during presentations of the most preferred and less preferred flickering lights alternately. Results showed that the effective duration of the autocorrelation function, tau(e), which represents a repetitive feature of the MEG alpha waves, becomes longer during the preferred condition. This reveals that the brain repeats a similar rhythm under preferred conditions. PMID- 11930176 TI - Genetic modifiers that aggravate the neurological phenotype of the wobbler mouse. AB - The autosomal recessive mutation wobbler of the mouse (phenotype WR; genotype wr/wr) causes muscular atrophy due to motoneuron degeneration with 100% penetrance on the standard Mus musculus laboratorius C57BL/6J background. In inter- and backcrosses with M. m. castaneus strain CAST/EI we have observed a variability in the severity of neurological symptoms. Approximately 15% of the WR (wr/wr) CAST/B6 hybrids were modified wobbler (WR*) mice defined by an aggravated neuromuscular phenotype with hindlimbs severely affected in addition to forelimbs. Histologically the overt WR* phenotype was paralleled by a caudally extended neurodegeneration in the ventral horn of the spinal cord with severe astrogliosis, and levels of acetylcholine receptor alpha-subunit mRNA in leg muscle much higher than in standard WR mice. Segregation analysis, using 68 polymorphic autosomal markers in a whole genome scan, revealed a major modifier gene locus, termed wrmod1, on chromosome 14. Individual recombination events in chromosome 14 consomic mice narrowed the wrmod1 candidate region to a 29 cM interval between D14MIT154 and D14MIT105, a region homologous to human chromosome 13q. Our analysis provides access to genes that modify neurodegeneration, the human counterparts of which may be responsible for the variable expression of hereditary spinal muscular atrophies. PMID- 11930177 TI - Lack of conscious recognition of one's own actions in a haptically deafferented patient. AB - How do we become aware of our own actions? This classical question is still a matter of debate: does consciousness depend on central efferent signals or derive from peripheral information? In this paper, we had the opportunity to study a haptically deafferented patient using a well-tested experimental paradigm where a cognitive conflict is produced between motor intention, proprioception and visual feedback. Our results show that the patient was able to solve the conflict and to generate accurate movements to a target in the absence of proprioceptive feedback and with very limited visual feedback from her movements. Yet, she could not report any conscious perception of the conflict and showed no conscious knowledge of her actual performance. We suggest that information derived from efferent processes cannot in themselves be a source for conscious experience about our own actions. PMID- 11930178 TI - Nanomolar amyloid beta protein activates a specific PKC isoform mediating phosphorylation of MARCKS in Neuro2A cells. AB - Myristoylated alanine-rich C kinase substrate (MARCKS), a protein associated with cell growth, neurosecretion and macrophage activation, is activated by protein kinase C (PKC) phosphorylation. We reported that amyloid beta protein (Abeta) activated MARCKS through a tyrosine kinase and PKC-delta in rat cultured microglia. Here we report that Abeta signaling pathway through a specific PKC isoform is involved in the phosphorylation of MARCKS in Neuro2A cells. Selective PKC inhibitors but not tyrosine kinase inhibitors significantly inhibited the phosphorylation of MARCKS induced by Abeta. Abeta selectively activated PKC-alpha among the four PKC isoforms localized in Neuro2A cells. PKC-alpha activated by Abeta directly phosphorylated a recombinant MARCKS in vitro, Translocation of PKC alpha from the cytoplasm to the membrane and accumulation of phospho-MARCKS in the cytoplasm were induced by Abeta. These results suggest involvement of a phosphoinositide signaling system through PKC-alpha in the phosphorylation of MARCKS in neurons, an event which may be associated with mechanisms underlying neurotrophic and neurotoxic effects of Abeta. PMID- 11930179 TI - Commissural glycinergic inhibition of bushy and stellate cells in the anteroventral cochlear nucleus. AB - Synaptic inputs from one cochlear nucleus (CN) to the other can play an important role in modulating the activity of CN neurons. Using the isolated whole brain preparation of the guinea pig, we tested the effects of electrical stimulation of the contralateral auditory nerve (AN) on intracellularly recorded and stained neurons of the anteroventral cochlear nucleus. Stimulation of the contralateral AN evoked only inhibitory postsynaptic potentials (IPSPs) in 63% of recorded neurons, including bushy and stellate cells. The latency of most IPSPs (88%) was in the range 3.3-7.6 ms, consistent with mono- and disynaptic transmission from the contralateral CN. The IPSPs had an average amplitude of 2.6 +/- 1.9 mV and were blocked by strychnine suggesting their glycinergic nature. These data, together with our similar findings in other CN subdivisions, indicate that principal cells of the CN contribute to binaural interactions at earliest stages of acoustic processing. PMID- 11930182 TI - Under-utilization of radionuclide therapy in metastatic bone pain palliation. PMID- 11930180 TI - Interleukin-1 beta is involved in the pressor response of amygdaloid neurons excited by sodium glutamate. AB - The effects of interleukin-1beta in the paraventricular nucleus or caudal arcuate nucleus on hypertensive and tachycardiac responses induced by excitation of the central amygdaloid nucleus were studied. We observed that microinjection of sodium glutamate into central amygdaloid nucleus resulted in hypertension and tachycardia. Microinjection of interleukin-1beta into paraventricular nucleus or caudal arcuate nucleus induced significant pressor and tachycardiac responses while pretreatment with microinjection of rabbit anti-rat interleukin-1beta antibody into bilateral paraventricular nuclei or arcuate nuclei attenuated the hypertensive response induced by microinjection of 40 nmol sodium glutamate into central amygdaloid nucleus. The above results suggest that the pressor effect of central amygdaloid nucleus is mediated by interleukin-1beta in paraventricular nucleus or arcuate nucleus. PMID- 11930183 TI - The pathology of hibernating myocardium. AB - Myocardial hibernation represents a protective mechanism of muscle preservation in the setting of atherosclerotic coronary artery disease. Long-standing myocardial hypoperfusion leads to diminished myocardial contractility that reverses with improved blood flow after revascularization. The morphologic changes in both animal models and humans are described. PMID- 11930184 TI - Detection and characterization of hibernating myocardium. AB - Since Tennant and Wiggers observed that coronary occlusion caused a reduction in cardiac contractile function, a lot has been written about the concept of hibernating myocardium. Known as the 'smart heart', hibernating myocardium is characterized by a persistent ventricular myocardial dysfunction with preserved viability, which improves with the relief of the ischaemia; this chronic downregulation in contractile function being a protective mechanism to reduce oxygen demand and thus ensure myocyte survival. This improvement usually results in an enrichment in the quality of life as well as enhanced ventricular function. In fact, it has been observed that the cardiac event rate in patients with viable dysfunctional left ventricular segments who are medically treated, is higher than the event rate in patients with comparable viability who are revascularized. Different degrees of histological alteration have been seen in hibernating myocardium, ranging from cellular de-differentiation (fetal phenotype) to cellular degeneration. Cellular de-differentiation has been associated with repetitive stunning. On the other hand, cellular degeneration (with more extensive fibrosis) has been associated with chronic low myocardial blood flow and a longer time to recovery after revascularization. These histological patterns may suggest an evolution from cellular de-differentiation to degeneration, which ends in scar formation if no revascularization is performed. In fact, several studies have described the clinical value of identifying and revascularizing hibernating segments as early as possible, to minimize fibrosis and morbidity from adverse events. Detection of hibernating myocardium still remains an important clinical problem. Imaging modalities to assess myocardial viability must differentiate potentially functional tissue from myocardium with no potential for functional recovery. These techniques fall into three broad categories: ventricular function assessment, myocardial perfusion imaging and myocardial metabolic imaging. PET imaging with fluorine-18 fluorodeoxyglucose (18F-FDG) and 11C-acetate, single photon emission computed tomography (SPECT) with thallium and 99mTc-sestamibi, dobutamine echocardiograpy, magnetic resonance imaging (MRI) and fast computed tomography (CT) have been used for this purpose. PET imaging, in both perfusion and glucose metabolic activity, has become a standard for myocardial viability assessment, however, similar information may be available from carefully performed studies with perfusion tracers alone. PMID- 11930185 TI - Assessment of myocardial viability by positron emission tomography. AB - Accurate assessment of myocardial viability is critical for identifying patients likely to benefit from coronary revascularization. Positron emission tomography (PET) has several advantages over single photon emission computed tomography (SPECT), including higher sensitivity and specificity, as well as the ability to measure myocardial blood flow and myocardial metabolism in absolute terms, which is important in understanding the pathophysiology of ischemic cardiomyopathy. The most commonly used PET tracer is [18F]2-fluoro-2deoxy-D-glucose (FDG). The dependence of ischemic myocardium on glucose metabolism makes FDG an ideal tracer in this setting. Studies have shown positive and negative predictive values for the detection of viable myocardium in the range of 48-94%, and 73-96%, respectively. FDG is superior to SPECT using thallium or technetium myocardial perfusion agents, as well as echocardiography with dobutamine infusion. FDG PET also provides important prognostic information. Patients with evidence of myocardial viability by FDG PET have fewer cardiac events and survive longer if revascularized compared to patients who are treated medically. This article will review myocardial metabolism, PET procedures and interpretive criteria, as well as problems and limitations. Data from the literature regarding diagnostic and prognostic information will also be summarized. PMID- 11930186 TI - Echocardiographic and magnetic resonance methods for diagnosing hibernating myocardium. AB - Hibernating myocardium refers to regions of impaired left ventricular function at rest due to coronary artery disease that is reversible with revascularization. The accurate identification and assessment of myocardial viability is a critical aspect of the management of the patient with coronary artery disease and left ventricular dysfunction. Several non-invasive methods exist to assist the clinician in distinguishing those patients with significant regions of hibernating myocardium from those who have non-viable scar. This is important not only to identify those patients who would most benefit from percutaneous intervention or surgery, but also to spare the latter group from the morbidity and mortality associated with a revascularization procedure that would provide little benefit. While nuclear medicine imaging is the most widely used means for evaluating myocardial viability, alternative modalities have emerged and have gained increasing acceptance in recent years. This article will review the echocardiographic and magnetic resonance imaging (MRI) methods that are currently available or under investigation to assess myocardial viability. These techniques include rest and stress echocardiography, myocardial contrast echocardiography, stress MRI, contrast-enhanced MRI and magnetic resonance spectroscopy (MRS). PMID- 11930187 TI - Clinical assessment in ischaemic cardiomyopathy. AB - Despite the trend of decreasing death rates attributable to ischaemic heart disease and stroke, the prevalence of heart failure and the resultant death rates in the United States have almost tripled between 1974 and 1994 [1]. Coronary artery disease is the commonest cause of heart failure in developed countries, accounting for up to 60% of cases. Advances in medical therapy, particularly the use of angiotensin-converting enzyme inhibitors and beta-blockers, have served to reduce morbidity and mortality in patients with left ventricular (LV) dysfunction due to coronary artery disease [2-5]. However, these improvements have been modest, and despite these therapies, patients with severe ischaemic cardiomyopathy continue to have a high mortality when treated medically. It is increasingly clear that the impaired LV function in these patients is not always an irreversible process. Traditionally, these observations have been made following demonstrable improvements in systolic function after coronary revascularization procedures. Diagnostic testing to evaluate the presence and extent of viable myocardium has therefore become an important component of the clinical assessment of patients with chronic coronary artery disease and LV dysfunction. PMID- 11930188 TI - Administered activity optimization in patients studied by equilibrium gated radionuclide ventriculography using pyrophosphate and technetium-99m. AB - Equilibrium gated radionuclide ventriculography is one of the most important fields of work in nuclear cardiology. The in vivo labelled method utilizing 99mTc and red cells labelled with pyrophosphate is one of the most widely used. In the present work determination of the optimum 99mTc activity was studied. A sample of 30 patients was divided into five groups which received various amounts of radioactivity: 303 MBq, 444 MBq, 617 MBq, 803 MBq and 1020 MBq. The image quality of each study was assessed qualitatively and by using a method based on the construction of an image quality discriminant function. The number of counts in regions of interest in the heart, liver, lung and spleen, and the background, were processed as variables. The best results for the selected variables were obtained with 803 MBq. PMID- 11930189 TI - Statistical parametric mapping in brain single photon computed emission tomography after carbon monoxide intoxication. AB - The purpose of this retrospective study was to assess regional cerebral blood flow in patients after carbon monoxide intoxication by using brain single photon emission computed tomography and statistical parametric mapping. Eight patients with delayed neuropsychiatric sequelae and ten patients with no neuropsychiatric symptoms after carbon monoxide intoxication were studied with brain single photon emission tomography imaging with 99mTc-hexamethyl-propyleneamine oxime. Forty four control subjects were also studied. We used the adjusted regional cerebral blood flow images in relative flow distribution (normalization of global cerebral blood flow for each subject to 50 ml x 100 g(-1) x min(-1) with proportional scaling) to compare these groups with statistical parametric mapping. Using this technique, significantly decreased regional cerebral blood flow was noted extensively in the bilateral frontal lobes as well as the bilateral insula and a part of the right temporal lobe in the patients with delayed neuropsychiatric sequelae as compared with normal volunteers (P< 0.005). In the patients with no neuropsychiatric symptoms, significantly decreased regional blood flow in the bilateral frontal lobes particularly on the left side was detected. There was a significantly decreased regional cerebral blood flow in the right frontal lobe and insula in the patients with delayed neuropsychiatric sequelae as compared to those with no neuropsychiatric sequelae. It is concluded that statistical parametric mapping is a useful technique for highlighting differences in regional cerebral blood flow in patients following carbon monoxide intoxication as compared with normal volunteers. The selectively reduced blood flow noted in this investigation supports the contention that the decrease following carbon monoxide intoxication may be prolonged and further worsen in the frontal lobe. In addition, the present study may help to clarify the characteristics of the pathophysiological alteration underlying delayed neuropsychiatric sequelae. PMID- 11930190 TI - Scintigraphy of acute inflammatory lesions in rats with radiolabelled recombinant human neutrophil-activating peptide-2. AB - Radiolabelled recombinant human interleukin-8 (IL-8) with its homologue neutrophil-activating peptide-2 (NAP-2) have been compared for imaging acute sterile inflammatory lesions in rats. 125I-IL-8 and 125I-NAP-2 were prepared by reaction with chloramine-T and injected intravenously into male rats bearing subcutaneous carrageenan abscesses in their left hindlimbs. Left hindlimb and right hindlimb activities were determined from serial total-body scintigrams between 1 h and 96 h post-injection as regional per cent injected activity corrected for physical decay (%IA). Time-activity curves for 125I-IL-8 and 125I NAP-2 in the carrageenan-containing left hindlimbs were similar in that both peaked at 1-3 h post-injection (IL-8, 4.9+/-0.5%IA; NAP-2, 4.8+/-1.9%IA) and decreased exponentially thereafter. However, while the lesioned-to-control limb activity ratio (L/C) for 125I-IL-8 only approximately doubled during the imaging period (1.7+/-0.3 at 1 h vs 3.7+/-1.0 at 24 h post-injection), L/C for 125I-NAP-2 more than tripled, rising from 1.5+/-0.4 at 1 h to 5.3+/-0.7 by 72 h post injection. It is concluded that while both radiolabelled IL-8 and NAP-2 may prove useful for clinical imaging, radiolabelled NAP-2 may provide better discrimination of inflammatory lesions from normal tissue at later times post injection. PMID- 11930191 TI - Development and validation of hydroxy ethyl starch kits for instant use in gastroesophageal reflux and gastric motility studies. AB - A reduction method based on stannous chloride is described to prepare hydroxy ethyl starch kits for gastrointestinal reflux and gastric motility studies. Following verification of the consistency of radiolabelling, in vitro experiments were carried out to validate 99mTc hydroxy ethyl starch as a liquid phase and solid phase gastric motility imaging radiotracer. Gastroesophageal reflux, liquid phase and solid phase studies were then conducted in 13 adult volunteers to examine the in vivo stability of the radiotracer. High labelling efficiency (>95% when prepared at neutral pH) was consistently achieved, which remained stable in conditions simulating gastric environment. Twelve of the 13 volunteers did not show absorption of any radioactivity from the gastro-intestinal tract. 99mTc hydroxy ethyl starch is a new agent suitable for gastroesophageal reflux and gastric motility studies. It is available in kit form and is a more 'physiological' agent than 99mTc sulphur colloid for preparing a solid radioactive meal. 99mTc hydroxy ethyl starch represents a true carbohydrate meal, and unlike 99mTc sulphur colloid, is easy to prepare and can easily be standardized to produce a standard vegetarian meal. PMID- 11930192 TI - Are patients informed when they consent to ERCP? AB - BACKGROUND: Only the British Society of Gastroenterology has published consent guidelines that are inclusive for endoscopic retrograde cholangiopancreatography (ERCP). Previous research has shown that there are variations in the information discussed with patients who are undergoing ERCP. PURPOSE: To examine the informed consent practices for ERCP in Ontario. METHODS: A self-report questionnaire was sent to ERCP endoscopists in Ontario, who were identified through a pre-existing database. The 14-item questionnaire included questions pertaining to the risks, benefits and alternatives discussed, how consent was obtained and whether the consent process was modified for patients older than 75 years. RESULTS: Of the 82 surveys sent, 36 responses were received, with three respondents indicating that they no longer performed ERCP; the total response rate was 40%. Ninety-four per cent of those who responded noted that they obtained written consent, and 6% obtained verbal consent. When discussing risks with their patients, 91% of respondents always mentioned pancreatitis, 88% always mentioned bleeding, 73% always mentioned perforation and 30% always mentioned the risk of infection; only 24% always mentioned the possibility of being allergic to the contrast agent, and 73% rarely or never mentioned death. When dealing with patients older than 75 years, 38% of respondents tended to be more brief in their explanations, 31% gave the same details in their explanation and 31% gave more detailed information than they gave to younger patients. Seventy-nine per cent mentioned the possibility of diagnostic failure and 82% mentioned the possibility of therapeutic failure, while only 27% mentioned the possibility of missing a diagnosis. CONCLUSIONS: Variability exists in terms of 'important information' given to patients undergoing ERCP. Standard informed consent guidelines specific to ERCP may help endoscopists uphold their responsibility to the patient, enhance patient understanding and reduce the risk of liability. PMID- 11930193 TI - Diagnosis of constipation in family practice. AB - BACKGROUND: Patients who complain of constipation to their family doctor may not be truly constipated. Variability in stool frequency and consistency, and perception of symptoms may lead to inaccurate patient reporting or diagnosis of constipation. OBJECTIVES: To determine whether patients visiting their family doctor with a complaint of, or diagnosed with, constipation fulfilled the Rome II criteria for functional constipation and had stool characteristics of constipation. METHODS: A random sample of Canadian family physicians were recruited to enroll a series of adults who complained of, or had received a diagnosis of, constipation during an office visit. Patients were advised of the survey. Those providing written consent were contacted by an independent research firm and forwarded a survey questionnaire that included the Rome II gastrointestinal questionnaire, questions regarding their medical history and questions regarding their demographics. Patients also completed a four-week daily diary recording their bowel habits using the Bristol Stool Form Scale, medication use and satisfaction with treatment. Questionnaire and diary responses were retrieved by telephone. RESULTS: One hundred eighty-four family physicians enrolled 311 patients, of whom 220 completed the questionnaire. Females comprised 79.5% of the sample and had a mean age of 54.2 years (males 61.6 years; P<0.05). According to the Rome II criteria, 37.3% had functional constipation and 46.8% had irritable bowel syndrome (IBS). Whole gut transit times estimated using the Bristol Stool Scale were similar among those with self-reported constipation, those with Rome II functional constipation and those with Rome II IBS (79.3 h, 85.8 h and 77.4 h, respectively). Almost half of the patients with IBS or functional constipation were taking a pain medication, while nearly one-fifth took antidepressants. Of the medications or remedies taken to treat constipation, patients rated 49.8% of the doses as satisfactory. CONCLUSIONS: A large proportion of Canadian primary care patients whose presenting complaint or diagnosis was constipation satisfied the Rome II criteria for IBS, with a smaller number defined as functionally constipated. IBS patients tended to be younger than those with functional constipation, and whole gut transit times did not differentiate IBS from functional constipation. Careful questioning of patients who complain of constipation may reveal constipating medication, diarrhea symptoms or IBS. PMID- 11930194 TI - Clinical experience with infliximab for Crohn's disease: the first 100 patients in Edmonton, Alberta. AB - OBJECTIVE: To determine whether the clinical efficacy and safety of infliximab in diverse clinical referral practices was similar to that seen in the randomized, controlled clinical trials. METHODS: Data were gathered from a review of charts of 109 consecutive patients with inflammatory and/or fistulizing Crohn's disease who received infliximab infusions. Responses were recorded based on the physician's global clinical assessment and classified as complete, partial or nonresponse. RESULTS: One hundred nine patients were treated with one to nine infusions of infliximab at a dose of 5 mg/kg and followed up for a median of 24 weeks (range one to 40 weeks). Fifty-four patients were treated for inflammatory disease, 38 for fistulizing disease and 17 for both. Clinical response occurred in 73% (17% complete response, 55% partial response). The clinical response rate did not vary relative to patient demographics, disease distribution, indication for infliximab, or the concomitant use of corticosteroids or immune modifiers. For those taking concomitant immune modifiers, the response rate was 75%. The median time to response was two weeks (range one to six weeks). The median duration of response was 12 weeks (range six to 88 weeks). Reduction or cessation of steroids was possible in 17 of 32 patients. Adverse events related to infliximab occurred in 7% of patients. These events were characterized as mild and did not require stoppage of infliximab therapy, except in one patient who had a treatable anaphylactic-like infusion reaction. CONCLUSIONS: The patient group in the present study realized significant clinical benefit, with minimal adverse effects, following treatment with infliximab. Clinical response rates paralleled those previously described in placebo controlled trials and retrospective clinical practice reviews. Nevertheless, the complete response rate (ie, remission) in this patient group was lower than that previously described. PMID- 11930195 TI - Small cell lung cancer with positive anti-Hu antibodies presenting as gastroparesis. AB - Lung cancer is the most common cancer in North America. Small cell lung cancer (SCLC) represents 15% to 25% of lung cancers. SCLC commonly relapses, resulting in a 3% to 8% five- year survival rate. The poor prognosis associated with SCLC is partly due to late diagnosis of the disease. Paraneoplastic syndromes can be early manifestations of SCLC. The potential benefit of early diagnosis has prompted investigations into markers of this disease. Some patients may present with predominantly gastrointestinal dysmotility symptoms that have no obvious explanation. Testing for anti-Hu antibodies, as a valuable marker of SCLC, should be considered in the investigation. A case of SCLC with positive anti-Hu antibodies presenting with intestinal pseudo-obstruction is presented. Gastrointestinal dysmotility as a manifestation of paraneoplastic syndrome is reviewed. PMID- 11930196 TI - ERCP: new standards, new challenges. PMID- 11930197 TI - Endoscopic evaluation of the small intestine. AB - Technological achievements in the area of endoscope design and development have resulted in instruments capable of advancing beyond the reach of simple gastroscopes. Such instruments, known as enteroscopes, form the bases of small bowel endoscopy. Recent widespread use of enteroscopes have contributed significantly to the understanding of small intestinal pathology and improved the ability to diagnose and treat patients with intestinal bleeding sources. PMID- 11930198 TI - Gallbladder polyps: epidemiology, natural history and management. AB - Polypoid lesions of the gallbladder affect approximately 5% of the adult population. Most affected individuals are asymptomatic, and their gallbladder polyps are detected during abdominal ultrasonography performed for unrelated conditions. Although the majority of gallbladder polyps are benign, most commonly cholesterol polyps, malignant transformation is a concern. The differentiation of benign from malignant lesions can be challenging. Several features, including patient age, polyp size and number, and rapid growth of polyps, are important discriminating features between benign and malignant polyps. Based on the evidence highlighted in this review, the authors recommend resection in symptomatic patients, as well as in asymptomatic individuals over 50 years of age, or those whose polyps are solitary, greater than 10 mm in diameter, or associated with gallstones or polyp growth on serial ultrasonography. Novel imaging techniques, including endoscopic ultrasonography and enhanced computed tomography, may aid in the differential diagnosis of these lesions and permit expectant management. PMID- 11930200 TI - [Intracellular potential changes in crucian carps Mauthner cells evoked by vagal sensory inputs]. AB - In the present experiments, microelectrode recording technique was used to explore the electrophysiological properties of Mauthner cells (M-cells) with stimuli applied to the right vagus. Direct stimulation of the right vagus evoked short-latency, long-lasting, graded, and complex postsynaptic potentials (PSPs) on both Mauthner cell bodies in crucian carp, which are stimulus strength- and frequency- dependent. The evoked PSPs reduced the amplitudes of the antidromic action potentials. After intramuscular injection of strychnine, both amplitude and averaged duration of the PSPs increased, on which two or more action potentials were superimposed. The results suggest that the afferent pathways from vagus to M-cells are mediated by several neuron chains with different numbers of relayed synapses, and they may be excitatory and/or inhibitory in nature. Therefore vagus input would modulate the excitability of the M-cells. PMID- 11930199 TI - [An exploration of animal behavior screen platform for novel gene function in central nervous system]. AB - For the purpose of large-scale screening of novel gene functions in mammalian nervous system, we have developed an animal behavior-monitoring platform employing antisense-oligo technology. Twenty genes of different categories were chosen from a low abundant gene (c)DNA sub-library of rat brain. Antisense oligo nucleotides of these genes were designed and synthesized according to the homologues of the genes in mouse for mouse behavior tests. These antisense oligos were injected into the lateral ventricles of mouse brain using a Hamilton micro syringe, with saline and oligos of scramble sequences as controls. These mice were tested with the following behavior model paradigms: metabolism, open field behavior, tail flick latency, and step-down test. Out of the 20 genes tested, 14 genes showed significant behavioral differences from the control groups at the level of P value less than 0.05 or 0.001 in different behavior animal models. PMID- 11930201 TI - [Suppression of glutamate neurotoxicity by 5-HT in hippocampal neurons]. AB - The effects of excess glutamate on the survival rate of cultured hippocampal neurons, the population spike (PS) of CA1 area of hippocampal slices and the Ca(2+) current of hippocampal neurons were observed in rats. Decreased survival rate of hippocampal neurons in the presence of excess glutamate was partially reversed by 5-HT, which was in association with a less decrease of PS magnitudes. The Ca(2+) inward current induced by glutamate was significantly diminished in the presence of 5-HT. It is suggested that 5-HT can inhibit excess glutamate induced neurotoxicity by suppressing the glutamate triggered calcium influx. PMID- 11930202 TI - [The biochemical properties of L-arginine/nitric oxide pathway in rat brain mitochondria]. AB - Biochemical property of L-arginine (L-Arg)/nitric oxide synthase (NOS)/nitric oxide (NO) system was observed in rat brain mitochondria. The results showed that there existed L-Arg transporters of high-affinity, low-transport and saturability in mitochondria membrane of normal rat brain. The maximum transport velocity (V(max)) and Michaelis constant (Km) were 5.87+/ -0.46 nmol/mg pro x min(-1) and 7.8+/- 0.56 micromol/L respectively. In normal condition, endothelium-type NOS was expressed in the mitochondria of rat brain, with its V(max) and Km being 2.7+/-0.3 nmol/mg pro x min(- 1) and 20.85+/-3.27 micromol/L, respectively. NO production was increased in a time-dependent manner and reached a maximum value at 60 min. These results suggest that the L-Arg transporters in MT membrane of normal rat brain were similar with Y(+) type amino acid transporters in cellullar membrane. Furthermore the NOS of mitochondria belongs to eNOS, whereas the L Arg/NOS/NO system may be a source of NO. PMID- 11930203 TI - [Increase in amount and affinity of adenosine receptor in rat hippocampal cellular membranes induced by cerebral ischemic preconditioning and its protective effects on the neurons]. AB - To determine the role of adenosine (ADO) receptor in the induction of brain ischemic tolerance, changes in amount and affinity of ADO receptor in rat hippocampal cellular membranes after transient ischemia were investigated using radioligand binding method. Ischemia for 6 min resulted in an apparent delayed neuron death (DND) in the hippocampus, while ischemia for 3 min did not cause DND. Preconditioning ischemia for 3 min could apparently decrease DND caused by ischemia for 6 min after the preconditioning at an interval of 1 d reperfusion. In correspondence with the histological changes, ischemia for 3 min caused an increase in amount and affinity of ADO receptor at 1 or 3 days after reperfusion (P<0.05 vs sham), while ischemia for 6 min caused a decrease in the amount and an increase in the affinity (P<0.05 vs sham). Compared with the rats suffering from ischemia for 6 min followed by reperfusion for 4 h and 1 or 3 d, the amount and affinity of ADO receptor increased in rats with preconditioning ischemia (3 min) 1 d before the ischemia for 6 min. The above results showed that cerebral ischemic preconditioning increased the amount and affinity of ADO receptor in hippocampal cellular membranes, and resisted the down-regulating of ADO receptor caused by severe ischemia, suggesting an important role of the increase in amount and affinity of ADO receptor in the induction of brain ischemic tolerance. PMID- 11930204 TI - [Effects of corticosterone on the activity of the neurons in the rostral ventrolateral medulla of rats]. AB - Spontaneous discharges of the rostral ventrolateral medulla (RVLM) neurons were extracellularly recorded by multi-barrelled microelectrodes in urethane anesthetized rats. A total of 145 neurons were recorded: 33 were cardiovascular, 31 were nociceptive modulating, and 81 were unknown functional. The cardiovascular neurons were electrophysiologically identified through activating baroreceptor reflex by electrical stimulation of the aortic nerve and by intravenous injection of phenylephrine. Of the 33 cardiovascular neurons, 25 (76%) increased in firing rate after iontophoretical application of corticosterone sulfate (CORT), and 8 (24%) failed to respond. Of the 31 nociceptive modulating neurons excited by noxious stimulation, the firing rate decreased in 19 (64%), increased in 2 (6%), and did not respond in 10 (30%) after iontophoresis of CORT. Of the remaining 81 unknown functional neurons, 32 (40%) were excited, 5 (6%) inhibited, and 44 (54%) were not affected by CORT. These results demonstrate that CORT may modulate the activities of the neurons in RVLM through fast non-genomical effect, suggesting that the mechanism of the fast actions of CORT may play an important role in integration of the cardiovascular, nociceptive modulating activity under some conditions such as stress. PMID- 11930205 TI - [Frequency-specific responses of human brain to peripheral transcutaneous electric nerve stimulation: a functional magnetic resonance imaging study]. AB - The purpose of the present investigation was to determine the responses of human brain to transcutaneous electric nerve stimulation (TENS) at different frequencies by functional magnetic resonance imaging (fMRI) examinations covering the whole brain of eleven healthy volunteers. Each subject received TENS at acupoints ST36 and SP6 of the left leg at the frequencies of 2 and 100 Hz. Frequency-specific responses were found in motor-related areas, thalamus, limbic system and associated cortex to stimulation of the two frequencies, while the primary somatosensory areas were activated by both. Therefore, it appears that 2 and 100 Hz TENS act through different neuro-pathways in the central nervous system. PMID- 11930206 TI - [Inositol 1,4,5-triphosphate receptors (IP(3)Rs) in myocardial nuclei involved in pressure overload-induced hypertrophy of rat heart]. AB - In order to clarify the importance of nuclear IP(3)Rs in the development of myocardial hypertrophy, a hypertensive rat model was established by abdominal aortic constriction, and velocity and isopyknic gradient centrifugation was employed to fractionate the cardiac nuclei. The maximal number of binding sites (B(max)) and dissociation constant (Kd) of IP(3) to the nuclear envelopes were measured by [(3)H] IP(3) binding assay. The existence of IP(3)Rs on myocardial nuclear envelope was confirmed. [Ca(2+)] inhibited [(3)H] IP(3) binding to its receptors in cardiac nuclear envelopes in a concentration-dependent way. Phosphorylation by CaM and endogenous PKC decreased B(max) of nuclear IP(3) receptors. B(max) and Kd of nuclear IP(3)Rs were increased by 1.217 (P<0.01) and by 2.149-fold (P<0.01) respectively in hypertrophic myocardium as compared with those of the control. The above results suggest that IP(3)Rs exist in myocardial nuclei and are down regulated by CaM, PMA and free Ca(2+). The increase of the binding sites of IP(3)Rs in the nuclear envelopes and the decrease of their affinity might play importment roles in the development of overload induction of cardiac hypertrophy. PMID- 11930208 TI - [Hemorrhagic shock induces changes in large-conductance Ca(2+) dependent K+ channel activity in mesenteric arterial smooth muscle cells of rats]. AB - The purpose of the present study was to detect changes in the activity of large conductance Ca(2+)activated K(+) channel (BK(Ca)) during hemorrhagic shock (HS) in mesenteric arterial vascular smooth muscle cells using inside-out patches. The HS rat model was produced with withdrawing blood from the femoral artery. The results showed that the BK(Ca) activity was inhibited with decreased open probability (P(o)) and single-channel conductance. The decrease of P(o) resulted from an increase of the close time constant (tau(cs)) in the compensatory stage of HS, i.e.the term of hyperresponse of the muscle to norepinephrine (NE) (HS lasted for about 40 min). During the decompensation stage of HS, i.e. the period of hyporesponse to NE (HS developed for 3 h), the BK(Ca) activity was enhanced with the increase in single-channel conductance and P(o) which was a result of a decrease in tau(cs). PMID- 11930207 TI - [Vasonatrin peptide attenuates hypoxia-induced increase in [Ca(2+)] (i) of culured rat cardiac fibroblasts]. AB - The purpose of this work was to test the hypothesis that vasonatrin peptide (VNP) can attenuate the growth-promoting effect by hypoxia in cardiac fibroblasts of cultured neonatal rats. Cultured fibroblasts were divided into four groups: control group, hypoxia group, VNP group and VNP+hypoxia group. The growth of cardiac fibroblasts was observed using MTT method and the incorporation of (3)H TdR, and the effect of VNP on the intracellular level of Ca(2+) was measured by means of interactive laser cytometry. It was found that hypoxia (2% - 3%) increased significantly the MTT optical density (OD) of cardiac fibroblasts (P<0.05 vs control group), but the increase was greatly attenuated in the VNP (10(- 6)mol/L) group and also the incorporation of (3)H-TdR in cardiac fibroblasts (P<0.05 vs hypoxia group). VNP (10(- 6)mol/L) also decreased the intracellular level of Ca(2+) which was increased by hypoxia (P<0.05) as compared with control and hypoxia group. These findings demonstrate that VNP can attenuate the hypoxia-induced growth-promoting effect in cardiac fibroblasts, which may be associated with the elevation of intracellular Ca(2+). PMID- 11930209 TI - [Changes in cyclin expression during proliferation and differentiation of CD34(+) cells derived from fetal liver induced by thrombopoietin]. AB - In order to elucidate the intrinsic mechanism underlying proliferation and differentiation of megakaryocytes during ontogenesis, CD34(+) cells were isolated from human fetal liver (FL) with a high-gradient magnetic sorting system (MACS) and were incubated in liquid suspension with 50 and 100 ng/ml of thrombopoietin (TPO) and in MegaCult(Tm) -C semi-solid culture system with 0, 12.5, 25, 50, 100, and 200 ng/ml of TPO. The cell number, colony number of CFU-Mk, platelet associated antigen phenotype, and DNA ploidy of CD41(+) cells were examined from d 0 to d 12 in culture. The expression patterns of cyclins B1, D1, and D3 were also analyzed by using immunoblot and flow cytometry. TPO stimulated proliferation of CD34(+) cells of FL from 1 x 10(5)/ml to 13.12 +/-4.06 10(5)/ml with 95% of CD41a(+) cells and 3% of CD34(+) cells after 12 d of culture. Most of the megakaryocytes (MKs) derived from FL were in 2 N ploidy class, and few in 4 N ploidy class, but no megakaryocytes ploidy class was higher than 4 N. The effect of TPO on the formation of CFU-Mk colonies from FL derived CD34(+) cells is shown in a dose-response curve. The expression of cyclin B1 increased progressively and the high level of cyclin B1 was maintained in FL CD34(+) cells induced by TPO during 12 d of culture. A high level of cyclin B1 appeared on FL derived MKs of G1 phase at d 12. The expression of cy-of cyclins D1 and D3 gradually increased in FL CD34(+) cells, which was induced by TPO during the initial 6-day incubation. Afterwards, the level of cyclins D1 and D3 decreased progressively, particularly in MKs which were in G2+M phases. These data suggest that (1) TPO induced proliferation and differentiation of FL derived CD34(+) cells through upregulation of cyclin B1 in G2+M phases and cyclins D1 and D3 in all phases of cell cycle, and (2) Continuing high level of cyclin B1 and decreases of cyclins D1 and cyclin D3 on MKs in G2+M phases may contribute to a retardation of MK endoreduplication. PMID- 11930210 TI - [Inhibition of platelet function by L-arginine.L-aspartate salt]. AB - The effects of L-arginine.L-aspartate salt (DR) on platelet aggregation, adhesion and release were investigated. Platelet aggregation induced by adenosine 5 diphosphate (ADP) was significantly inhibited (P<0.01) by intravenous injection of DR (15 mg/kg) in rats or oral administration (15 mg/kg) in rabbits, the inhibitory effect on rabbit platelet aggregation lasting for more than 8 h (P<0.01). Platelet aggregation induced by ADP, collagen or thrombin in rats was all markedly inhibited by 7.5, 15 or 30 mg/kg of DR (bid for 3.5 d, ig, P<0.01). Platelet adhesion to foreign objects was inhibited by 30 mg/kg of DR (ig). Bleeding time in rat tails was prolonged by 30 mg/kg of DR (P<0.05). Furthermore, PGI(2) released from the vascular wall was increased in DR-treated rats (P<0.05), however, TXA(2) released from platelets was not affected. These data demonstrate the inhibitory effect of DR on platelet function, suggesting that its action target may be different from that of acetylsalicylic acid, and that the increase of PGI(2) release may be responsible partly for this effect. It is suggested that DR may probably be used as a new agent for regulating platelet function. PMID- 11930211 TI - [Properties of peroxynitrite-induced relaxation in rabbit pulmonary artery in vitro]. AB - Vasodilatory properties of exogenous peroxynitrite (ONOO(-)) and effects of endothelial cells on ONOO(-) induced relaxation were investigated in isolated rabbit pulmonary arterial rings (PARs). In pre-contracted PARs, ONOO(-) gave rise to vasodilation in a dose-dependent manner, which was significantly higher than that of decomposed ONOO-. In contrast, relaxation of PARs to ONOO(-) was lower, as compared with sodium nitroprusside (SNP) or acetylcholine(ACh). ONOO(-) induced more significant relaxation in denuded endothelial PARs than in intact endothelial PARs. Relaxation of PARs to repetitively administered ONOO(-) appeared progressively decreased. Under this experimental condition, relaxation of PARs to ACh remained unchanged after administrating ONOO(-). These results suggest that ONOO(-) causes weak relaxation in pulmonary artery, which is down regulated by endothelium and is of tachyphylaxis. PMID- 11930212 TI - [Current responses mediated by endogenous GABA(B) and GABA(C) receptors in Xenopus oocytes]. AB - The properties of GABA-activated current in Xenopus oocytes and its underlying mechanism were studied using the two-electrode voltage-clamp technique. External application of GABA (10(-10) - 10(-3)mol/L) induced a concentration-dependent outward current in a proportion of oocytes (35.5%, 55/155). Selective GABA(A) receptor antagonist bicuculline (10(-5) mol/L) did not block the GABA-activated current (n=6). However, 2-hydroxysaclofen (10(-4) mol/L), a GABA(B) receptor antagonist, reversed the GABA-activated outward current to an inward current (n=9), which was abolished completely by application of I4AA (10(-5) mol/L), a GABA(C) receptor selective antagonist (n=6). In 20% (12/60) of oocytes, application of baclofen (10(-10) - 10(-4) mol/L), a GABA(B) receptor agonist, also induced a concentration-dependent outward current 2-Hydroxysaclofen at the concentrations of 3 x 10(-6), 3 x 10(-5) and 3 x 10(-4) mol/L blocked the baclofen(10(-5) mol/L)-activated outward current by (6.3+/-3.2) %, (44.1+/-2.2) %, and (86.0+/-1.6) %, respectively (n=6). The reversal potential for baclofen activated current was around 96.8+/-7.2 mV (n=6), and the baclofen-activated current could be blocked by TEA (n=5) and Ba(2+) (n=5). These results suggest that there exist endogenous GABA receptors, GABA(B) receptors mediating a slow and sustained outward current and GABA(C) receptors mediating an inward current in follicular Xenopus oocytes. PMID- 11930213 TI - [Effects of murine bone marrow endothelial cell conditioned medium in combination with FL and TPO on the growth of HPP-CFC and CFU-GM]. AB - In the present study, the effects of murine bone marrow endothelial cell conditioned medium (ECM) combined with flt3 ligand (FL) or/and thrombopoietin (TPO) on the proliferation of HPP-CFC and CFU-GM were investigated. Both ECM and the concentrated retentate of ECM (MW>10 kD) promoted the growth of CFU-GM and HPP-CFC, and this promoting effect was further enhanced by addition of FL or TPO. Using reverse transcriptase-polymerase chain reaction (RT-PCR) technique, the expression of FL and TPO mRNA was not found in murine bone marrow endothelial cells. PMID- 11930214 TI - [Preliminary separation and identification of the neurotrophic substances from dorsal spinal cord of chicken embryos]. AB - Extracts of spinal dorsal horn and ventral horn of chicken embryos hatched for 9 days were separated by Sephadex G-75 gel filtration, and different chromatographic fractions of DI and DII, VI and VII were obtained. The neurite outgrowth promoting effects of the various fractions were assayed by the dorsal root ganglion (DRG) culture in vitro. It was shown that DII, but not DI, fraction of the dorsal horn extract exhibited a significant neurite-outgrowth promoting effect on DRG. By means of SDS-PAGE, the molecular weight of DII fraction ranged from 61 to 15 kD. VI and VII fraction of ventral horn showed no obvious effect on neurit-outgrowth of DRG in vitro. PMID- 11930215 TI - Changes in the contents of glycogen and lactate in the brain and blood during hypoxic preconditioning. AB - Mice were randomly divided into groups H4 (hypoxic preconditioning group with repetitive hypoxic exposures for four runs), H(1) (hypoxic control group with exposure to hypoxia for one run) and H(0) (normal control group with no exposure to hypoxia). Glycogen content of whole brain of group H4 was found to be significantly higher than that of groups H(1) and H(0). The glycogen content in telencephalon, diencephalon and pons of group H4 was markedly higher than that in the corresponding areas of groups H(1) and H(0). Glycogen content of whole brain in group H(1) was markedly lower than that in group H(0), whereas no significant difference was seen in these brain subregions between groups H(1) and H(0). Brain lactate contents of groups H4 and H(1) did not show significant difference, though they were significantly higher than that of group H(0). Blood lactate content of group H4 was significantly lower than those of groups H(1) and H(0). The results above indicate that the concomitant increase of glycogen and decrease of lactate in the brain are due to the participation of aerobic metabolism during hypoxic preconditioning or the formation of tolerance to hypoxia. PMID- 11930216 TI - [Apoptosis during placentation]. AB - Apoptosis is a normal physiological phenomenon. During the process of implantation, there are a large number of cells at the implantation site undergoing apoptosis, which has been suggested to play an important role in the regulation of endometrium decidulization and trophoblast invasion. Apoptosis mediated by Fas/FasL system may also be associated with maternal immunotolerance to the fetus. In this review, the common pathway of apoptosis and the regulation in the placenta are described. PMID- 11930217 TI - Rapid activation of p38 mitogen-activated protein kinase by corticosterone in PC12 cells. AB - The present study using immunoblot showed that corticosterone (B) could induce a rapid activation of p38 in PC12 cells. The dose and time response curves w ere bell shaped with a maximal activation at 10-9 mol/L and 15 min respectively. The activation was not affected by steroid nuclear receptor antagonist RU38486. Bovine serum albumin coupled B (B-BSA) could induce phosphorylation of p38. Tyrosine kinase inhibitor genistein failed to block the phosphorylation, a fact suggesting that the tyrosine kinase activity is not involved in the pathway. On the other hand, phorbol 12-myristate 13-acetate (PMA), a protein kinase C (PKC) activator, could mimic the actions of B, while G 6976, a PKC inhibitor, could completely abolish the phosphorylation induced by B. These results clearly demonstrate that B activates p38 MAPK readily via a putative membrane receptor through a PKC-dependent pathway. PMID- 11930219 TI - [Effect of interleukin-2 on intracellular calcium transients in rat ventricular myocytes]. AB - In the present study, we investigated the effect of interleukin-2 (IL-2) on the intracellular calcium in enzymatically isolated ventricular myocytes with the use of the spectrofluorometric techniques. It was shown that IL-2 (2.5 200 U/ml) depressed electrically induced Ca(2+) (i) transients of ventricular myocytes in a dose dependent manner. IL-2 (200 U/ml) did not alter the caffeine releasable pool of Ca(2+). Pretreatment with the non selective opioid antagonist naloxone (10(-8)mol/L) or a specific kappa opioid antagonist nor binaltorphimine (nor-BNI, 10(-8) mol/L) abolished the inhibitory effect of IL-2 (200 U/ml) on the Ca(2+) (i) transients of cardiomyocytes, whereas the specific delta opioid antagonist naltrindole (10(-6) mol/L) did not abolish the inhibitory effect. The effect of IL-2 (200 U/ml) was also abolished after pretreatment with pertussis toxin (PTX, 5 mg/L) as well as phospholipase C (PLC) inhibitor U73122 (5 10(-6) mol/L), but not by tyrosine kinase inhibitor genistein (10(-4) mol/L). It is concluded that the depressant effect of IL-2 on the Ca(2+) (i) transients of isolated ventricular myocytes is mainly mediated by cardiac kappa opioid receptor pathway including a PTX sensitive Gi-protein and PLC, but not by tyrosine kinase. PMID- 11930218 TI - [Modification of membrane lipid of human umbilical vein endothelia cells after direct lipopolysaccharide injury]. AB - Our previous study showed that the membrane viscosity of human umbilical vein endothelial cells (HUVECs) increased with lipopolysaccharide (LPS) concentration after direct LPS damaging, implying that LPS could change the membrane structure and composition of HUVECs. The purpose of the present study was to investigate the modification of HUVECs membrane lipid by LPS under direct LPS injury to HUVECs. Phospholipid components of HUVEC membrane were determined with high performance capillary electrophoresis (HPCE), and HUVECs were directly damaged by LPS of different concentrations: 0, 0.313, 0.625, 1.25, 2.5, 5 and 10 microgram/ml for 3 h, and by 0.625 microgram/ml for 1, 3, 6, 12, 24 and 48 h. The results show that total phospholipid content of HUVECs increased with the increase of LPS concentration at 3 h, and decreased with the duration of LPS effect at 0.625 microgram/ml. Phosphatidylinositol (PI) and phosphatidylcholine (PC) contents increased with the increase of LPS concentration and decreased with the duration of LPS effect; LPS contents and its effect duration had slight effect on sphingomyelin (SM), phosphatidylethanolamine (PE) or phosphatidylserine (PS) contents. The LPS activating effect of HUVEC membrane lipid showed a typical property of enzymatic dynamics. These experimental results indicate that LPS directly activates the metabolism of HUVEC membrane lipid and induces the modification of HUVEC membrane phospholipid, which implies that the mechanism of the impairment of HUVECs by LPS may be related to the membrane structure of HUVECs and the metabolism of HUVEC membrane lipid. PMID- 11930220 TI - [Influence of interleukin-10 on nitric oxide /nitric oxide synthase system of the aorta]. AB - To study the influence of the inflammatory factor---interleukin-10 on nitric oxide (NO) and nitric oxide synthase system of rat aorta, Griess assay, production of (3)H-citrulline and Western blot were used to determine the release of NO, and the activity and expression of nitric oxide synthase, respectively. The results showed that lipopolysaccharide (LPS) stimulated the activity of inducible nitric oxide synthase (iNOS) and the release of NO. 10(-10) 10(-8) g/ml of IL-10 induced the activity and expression of endothelial nitric oxide synthase (eNOS), but not of iNOS. 10(-9) 10(-8) g/ml of IL-10 also downloaded the release of NO, and the activity and expression of iNOS induced by LPS, while 10(-7) g/ml of IL-10 significantly stimulated the activity and expression of iNOS but not eNOS. In summary, IL-10 presents a duple role in NO/NOS system. On the one hand, IL-10 inhibits the activity and expression of iNOS induced by inflammatory factor; on the other hand, IL-10 stimulates the release of endothelial NO. PMID- 11930221 TI - [Identification of genes related to cell phenotypic transition by differential display analysis]. AB - To identify the genes that are differentially expressed during the phenotypic transition from vascular adventitial fibroblasts to myofibroblasts, the adventitial fibroblasts were cultured from rat thoracic aorta, and myofibroblasts were obtained by treatment of fibroblasts with TGF-beta1. Differential display PCR (DD-PCR) was used to screen for differentially expressed genes by comparison of mRNA extracted from the two cell populations. Bands upregulated or downregulated on DD gels were excised, reamplified, cloned and sequenced. DD results were verified by quantitative PCR and Northern blot analysis.Antisense oligonucleotide was transfected to study the effect of osteopontin on migration of AF. Differential display showed a significant difference in gene expression profile between the two cell types. A transcript that was downregulated in myofibroblasts showed high DNA sequence homology to part of the gene for NADH dehydrogenase subunit 5. An upregulated transcript showed significant sequence homology to osteopontin gene. Quantitative PCR and Northern blot analysis confirmed the DD results. Among the other differential bands detected, 4 candidate sequences showed no homology to the known genes. The AF numbers of migration were significantly decreased by use of OPN antisense oligonucleotide. This study suggests that the downregulation of gene encoding NADH dehydrogenase subunit 5 and upregulation of osteopontin gene and several other unknown genes may be involved in the phenotypic transition of adventitial fibroblasts to myofibroblats. Inhibition of the expression of OPN may play an important role in the process of vascular remodeling. PMID- 11930222 TI - [Changes in Galphaq/11 mediated signal transduction pathway in the aorta of two kidney one clip hypertensive rats]. AB - The objective of this study was to investigate the role of signal transduction pathway mediated by Galphaq/11 of the aorta in blood pressure regulation during the development of two-kidney one clip (2K1C) Glodblatt hypertension in the rat. Levels of Galphaq/11-subunit and extracellular signal regulated kinase 1/2 (ERK1/2) of the aorta were measured by Western blot analysis at week 1, 2, 4 or 8 after operation. Phospholipase C (PLC) activity of the aorta was measured by using (3)H phosphatidylinostol (4,5) bisphosphate as a substrate. Both systolic and diastolic blood pressure increased markedly in 2K1C rats at week 2, 4 or 8, whereas, the increase in Galphaq/11 and ERK1/2 expression of the aorta began at week 1 after operation (increased by 57.53% and 40.16%, respectively), and maintained at a high level during week 2 8 compared with time matched controls (P<0.01). PLC activities in the aorta were also increased significantly in 2K1C groups at week 4 and 8 compared with the time matched controls (P<0.05). The results indicate that Galphaq/11-mediated signal transduction pathway of the aorta is activated in 2K1C hypertension, and may contribute to the initiation and maintenance of renal hypertension. PMID- 11930223 TI - Renal ischemia-induced Fos expression in catecholaminergic neurons of rats. AB - The present study was undertaken to define whether the renal artery occlusion (RAO) would activate the catecholaminergic neurons in the brainstem nuclei by double immunohistochemical method for detecting Fos and tyrosine hydroxylase. The results are as follows: (1) while the basal expression of Fos was relatively low in the brainstem, RAO was capable of inducing a robust Fos-like immunoreactive neurons in the nucleus tractus solitarius (NTS), area postrema (AP), nucleus paragigantocellularis lateralis (PGL) and locus coeruleus (LC); (2) numerous catecholaminergic neurons in NTS, AP, PGL and LC could be activated by RAO as shown by Fos expression; and (3) these responses to RAO were significantly attenuated by pretreatment with an adenosine receptor antagonist 8 phenyltheophylline. The results suggest that RAO can activate a large number of neurons including some catecholaminergic neurons in the brainstem nuclei. Such effects of renal ischemia may be attributed to RAO-induced adenosine release from the kidney which subsequently activates renal afferents. PMID- 11930224 TI - Differential responses of regional vascular beds to local injection of agmatine in rats. AB - In 66 anaesthetized rats, the effects of local injection of agmatine on femoral, renal, and mesenteric vascular beds were investigated respectively by constant flow perfusion method. The results are as follows. (1) Agmatine (0.1, 0.5, 1 mg/kg) increased the perfusion pressure (PP) of femoral vascular bed in a dose dependent manner. The effect of agmatine (1 mg/kg) was completely blocked by pretreatment with idazoxan (0.5 mg/kg), an antagonist for imidazoline receptors (IR) and alpha(2)-adrenergic receptors (alpha(2)-AR), and yohimbine (1 mg/kg), a selective alpha(2)-AR antagonist. (2) Agmatine also increased the PP of renal vascular bed in a dose-dependent manner, and agmatine at high dose (1 mg/kg) caused a biphasic increase of PP in renal vascular bed. Idazoxan blocked these effects completely, while yohimbine led the agmatine induced effect to a decrease in the PP of renal vascular bed. (3) Agmatine decreased the PP of mesenteric vascular bed in a dose-dependent manner, an effect which was completely blocked by idazoxan, but unaffected by yohimbine. From the results obtained, it is concluded that agmatine differentially affects the vascular tone in the femoral, renal, and mesenteric vascular beds. PMID- 11930225 TI - [Responses of afferent unit of the caudal nerve of diabetic hyperalgesic rats to sympathetic efferent stimulation]. AB - Responses of afferent unit to sympathetic stimulation (SS), intravenous injection of noradrenaline (NA) and phentolamine in the caudal nerve of diabetic rats were investigated. The results showed that the discharge frequencies of C and Adelta units with spontaneous discharges were increased in diabetic hyperalgesic rats after SS, and these spontaneous discharges were eliminated by adrenergic antagonist. The C (6/21) and Adelta (19/81) units with no spontaneous discharges of diabetic hyperalgesic rat turned from silent state into active state during SS; although SS did not elicit afferent discharges of the C mechanical receptive units (C-M), it elicited afferent discharges of a part of C mechano heat units (C-MH) and C polymodal units (C-Pol); afferent discharges of some of the Adelta mechanical receptive units (Adelta-M) and Adelta mechano heat units (Adelta-MH) were also elicited by SS. The latencies of the C and Adelta units responses upon SS were not equal, but no less than 5 s. SS elicited neither afferent discharges from Abeta mechanical receptive units of diabetic hyperalgesic rat, nor receptive units of all types in the control rat. The C(3/8) and Adelta (4/12) units of diabetic hyperalgesic rat were activated by intravenous injection of NA. The present data suggest that NA released from sympathetic nerve terminals excites C and Adelta units of diabetic hypesthesic rat, which may be a peripheral factor in hyperalgesia and paresthesia of diabetic rats. PMID- 11930226 TI - Changes in glycine content in mouse brain during hypoxic preconditioning. AB - Glycine content of the whole brain and subregions was measured by high pressure liquid chromatography (HPLC) in mice during their repetitive exposures to hypoxia. The contents of glycine in whole brain, diencephalon, and hippocampus and brain stem was significantly increased as the animals tolerance to hypoxia was increased. The results suggest that glycine, as an inhibitory neurotransmitter in the brain, may positively contribute to the development of hypoxic preconditioning. PMID- 11930227 TI - The role of central arginine vasopressin in corticotropin releasing hormone induced fever in rats. AB - The purpose of the present study was to investigate the role of central arginine vasopressin (AVP) in corticotropin releasing hormone (CRH) induced fever in the rat. Guide cannulae were inserted into the third ventricle and placed over the ventral septal area (VSA). The content of arginine vasopressin in the VSA of the brain was determined by radioimmunoassay. Colon temperature was monitored in lightly restrained rats by insertion of a catheter mounted thermistor probe 5 cm in the rectum. The results demonstrated that intracerebroventricular (icv) injection of CRH increased AVP level in the VSA and the colonic temperature of the rats. Microinjection of AVP V(1) antagonist into the VSA 10 min before CRH administration significantly enhanced CRH-induced febrile response, while AVP V(1) antagonist itself did not have a significant effect on the colonic temperature. Furthermore, injection of AVP into the VSA 5 min before CRH administration (icv) suppressed the fever evoked by CRH. These findings suggest that CRH is an important factor that stimulates the release of AVP in the VSA during fever, and endogenous AVP in the VSA has an antipyretic action on the CRH induced fever. PMID- 11930228 TI - Correlation between mitochondrial membrane potential and neurotoxic effect of corticosterone on primary cultured hippocampal cells. AB - Experiments were carried out to study the toxic effect of corticosterone (CORT) on primary cultured hippocampal cells and its relationship with mitochondrial membrane potential (MMP). Hippocampal cell survival and MMP were assessed by MTT and laser scanning confocal microscopy respectively. The results indicated that the survival rate of hippocampal neurons and astrocytes was dose dependently reduced in the hypoglycemic (3 5 mmol/L) and serum free medium for 24 h. With the same concentration of CORT, the toxic effect of CORT on hippocampal neurons was more serious than that on astrocytes. A supplement of high concentration of glucose (25 mmol/L) greatly improved the survival rate of hippocampal neurons cultured in the hypoglycemic and serum free medium. The disruption of MMP was induced by CORT (10( 6) mol/L) in hippocampal neurons cultured in hypoglycemic and serum free medium and antagonized by high concentration of glucose. It is therefore suggested that hippocampal neurons are more sensitive to the toxic effect of CORT in comparison with astrocytes. Neurotoxic effect of CORT can be reduced by high concentration of glucose. CORT-induced reduction of MMP may be one of the mechanisms for its neurotoxicity. PMID- 11930229 TI - [Increased cellular proliferation in BEL-7402 hepatoma cells transfected by human hepatic stimulator substance gene]. AB - A hepatotropic factor originally identified in weanling or regenerating rat livers, known as hepatic stimulator substance (HSS), is characterized by promoting markedly liver regeneration after partial hepatectomy. Although HSS gene and the gene product have been described recently, the cellular mechanism of HSS and its genetic function remain obscure. In our previous studies, human HSS (hHSS) was demonstrated to promote the growth of hepatoma cells and phosphorylate the mitogen-activated protein kinase (MAPK). In this study, we reported the growth features of human hepatoma cells response to hHSS gene transfection. The hHSS eukaryotic vector was transfected to BEL-7402 hepatoma cells and the expression of hHSS was analyzed with Northern and Southern blot. The results showed that the HSS recombinant construct was functionally expressed in the target cells as analyzed with Northern blot and reverse transcriptase polymerase chain reaction (RT-PCR). The growth potential of HSS transfected hepatoma cells was markedly enhanced as compared with the non transfected cells. The S-phase of the hHSS-transfected cells increased by 61.5% than that of the non-transfected cells as shown by flow cytometry. Moreover, MAPK phosphorylation, one of the most important mitogenic indexes of growth signal cascade, was profoundly activated at sites of Thr202/Tyr204 due to HSS gene transfer. Based on these results, it is concluded that the introduction of HSS gene into hepatoma cells might be able to stimulate directly the cellular growth in vitro, allowing a possibility of reevaluation of HSS mechanism in intact cells. PMID- 11930230 TI - [Role of nitric oxide in cholecystokinin octapeptide alleviation of tumor necrosis factor alpha induced changes in rabbit pulmonary arterial reactivity]. AB - To explore the mechanism underlying cholecystokinin octapeptide (CCK-8) induced attenuation in pulmonary arterial hypertension (PAH) in endotoxic shock (ES), the effect of CCK-8 on the changes in rabbit pulmonary arterial reactivity induced by tumor necrosis factor-alpha (TNF-alpha) was observed with isolated arterial ring technique and by examination of nitric oxide synthase (NOS). The contractile response to 10(-6) mol/L phenylephrine (PE) and the endothelium dependent relaxation response to 10(-6) mol/L acetylcholine (ACh) were not affected by TNF alpha (4000 U/ml) after incubation for 2 h; the relaxation response was decreased significantly when the incubation was prolonged to 7 or 14 h, which, however, could be reversed by a concomitant exposure to CCK-8 (0.5 microgram/ml), but the incubation of pulmonary arterial rings with CCK-8 (0.5 microgram/ml) alone did not bring out any contractile responses. The endothelium dependent relaxation response to 10(-6) mol/L ACh was restored by L arginine in the TNF-alpha group which had been incubated for 7 h, but was not affected by AG in each group, while the contractile response to 10(-6) mol/L PE increased significantly in the TNF alpha group. The relaxant response to 10(-6) mol/L ACh changed into a contractile response after preincubation with L-NNA in each group, while the contraction response to 10(-6) mol/L PE increased significantly. The NOS activity increased in the TNF-alpha and the TNF alpha+CCK-8 groups, while no significant difference was observed between the vehicle and the CCK-8 groups. These results suggest that CCK-8 prevents TNF-alpha induced impairment in endothelium dependent relaxation response, and the effects of both CCK-8 and TNF-alpha are related to NO. PMID- 11930231 TI - Promotion of pulmonary fibroblast proliferation and apoptosis by sodium nitroprusside. AB - The effects of nitric oxide (NO) donor sodium nitroprusside (SNP) on the proliferation and apoptosis and on Bcl-2, Bax and p53 proteins of pulmonary fibroblasts were investigated by using MTT cleavage assay, agarose gel electrophoresis and flow cytometric analysis. The results showed increases in the optical density (550 nm) of MTT cleavage assay, the number of cells and the proliferation index (PI), in comparison with the control. The number of apoptotic cells was also increased, though the percentage of apoptotic cells was too low to reveal oligonucleosomal fragmentation of characteristic ladder pattern, which is associated with apoptosis. In the meantime, the level of Bcl-2 decreased and that of Bax increased, while the p53 remained unchanged. These results suggest that exogenous NO has a dual effect on proliferation and apoptosis; and the action of NO on pulmonary fibroblasts is mainly proliferative. Down regulation of Bcl-2 and up regulation of Bax are implicated in the molecular mechanisms of this action. PMID- 11930232 TI - The role of MDR1 gene in volume-activated chloride currents in pigmented ciliary epithelial cells. AB - The role of multidrug resistance (MDR1) gene in the activation of volume activated chloride currents in bovine pigmented ciliary epithelial (PCE) cells was investigated by the patch-clamp technique, the antisense approach, the immunofluorescent technique and the confocal microscopy. PCE cells express P glycoprotein (P-gp, the product of MDR1 gene). An MDR1 antisense oligonucleotide suppressed MDR1 expression (93% reduction of P-gp immunofluorescence), delayed the activation of a volume-activated chloride current (latency prolonged by 109%), reduced the activation rate by 62% and decreased the peak value of the current by 56%. The transfection reagent lipofectin and the mismatch control oligonucleotide did not significantly affect the current. The data indicate that the volume-activated chloride current is associated with the endogenous expression of MDR1 gene in PCE cells. PMID- 11930233 TI - [Inhibition of moderate hypoxia-induced protein synthesis by vasonatrin peptide in cultured neonatal rat cardiomyocytes]. AB - The present work was to investigate the effects of vasonatrin peptide (VNP) on cardiomyocyte protein synthesis induced by moderate hypoxia. In cultured neonatal rat cardiomyocytes, MTT methods, total protein measurement and (3)H-leucine incorporation were used to calculate the cell number and measure the protein synthesis of cardiomyocytes. Furthermore, radioimmunoassay was undertaken to observe the effects of VNP on the intracellular levels of cAMP, cGMP and the concentration of endothelin (ET) in the culture medium. The results showed that both the cell number and protein synthesis decreased with severe hypoxia for 24 h. In contrast, under moderate hypoxia, cardiomyocyte hypertrophy developed; the protein synthesis as evidenced by total protein content and 3H-eucine incorporation increased significantly. VNP reduced cardiomyocyte protein synthesis induced by moderate hypoxia in a dose-dependent manner. Furthermore, VNP increased the intracellular level of cGMP and decreased the concentration of ET in the culture medium under moderate hypoxia, but had no effect on the level of cAMP. These results suggest that VNP inhibits moderate hypoxia-induced protein synthesis in cultured neonatal rat cardiac myocytes. This effect is mediated, at least in part, by an increase in intracellular cGMP, a reduction in synthesis, and/or a release in ET of cardiomyocytes. PMID- 11930234 TI - [Increased resistance against oxidant-induced injury in the rat vascular smooth muscle cells transfected with human heme oxygenase-1 gene]. AB - The heme oxygenase-1 (HO-1), a rate-limiting enzyme in heme metabolism, has been recently defined as a novel stress-stimulated protein, since the intracellular expression of HO-1 in response to various stimuli as oxidation, ischemia and endotoxin injury has been proved to be able to protect the cells from damage. In this study, a retroviral vector containing human HO-1 gene was constructed and transfected to rat vascular smooth muscle cells (VSMCs). Using Southern and Northern blot analyses, the integration and mRNA expression of HO-1 gene in the transfected cells were confirmed. The profound protein expression of HO-1 as well as HO enzyme activity in the transfected cells increased by 1.8-fold and 2.0-fold respectively as compared with the non-transfected cells. It was found that the HO 1 transfected-VSMCs presented dominant resistance to toxicity produced by exposure to H2O2, as a significant protective effect of HO-1 marked by cell survival and LDH leakage was observed when 200, 400 and 600 micromol/L of H2O2 were used. The protection of HO-1 rapidly declined after the transfected-VSMCs were pretreated 24 h with an HO-1 specific inhibitor (ZnPP-IX). The results of this investigation suggest that the functional expression of HO-1 gene within VSMCs raises an alternative ability to protect the vascular cells against active oxygen injury. PMID- 11930235 TI - 17beta-Estradiol inhibits vascular smooth muscle cell proliferation and c -fos expression: role of nitric oxide. AB - Rat vascular smooth muscle cells (VSMC) were used to study the effect of 17beta estradiol (E(2)) on cellular proliferation (cell counting), DNA synthesis ((3)H thymidine incorporation), MTT, c -fos mRNA expression and nitric oxide (NO) release. The results obtained showed that E(2) (10(-12) 10(-8) mol/L) induced NO release from VSMC in a concentration-dependent manner; 10(-8) mol/L E (2)significantly inhibited VSMC cellular proliferation and DNA synthesis induced by 10% FCS and 10(-7) mol/L ET-1, which was obviously reversed by 10(-7)mol/L tamoxifen and 10(-6) mol/L L-NAME; after a pretreatment for 24 hours, 10(-8)mol/L E(2) significantly inhibited VSMC c -fos mRNA expression induced by 10(-7)mol/L ET-1, which was also obviously reversed by 10(-6) mol/L L-NAME. These results suggest that the inhibitory effects of E(2) on VSMC cellular proliferation and c fos mRNA expression are closely related with NO release in VSMC, which is, at least, partly medicated by ER. PMID- 11930236 TI - [Prokaryotic expression and purification of human hepatic stimulator substance]. AB - To explore the possibility of prokaryotic expression of human hepatic stimulator substance (hHSS), hHSS gene was inserted in the downstream of glutathion S transferase (GST) in a pET-42a expression vector and recombinant GST-hHSS fusion protein was expressed under IPTG induction in BL-21(DE3) cells. The recombinant HSS was purified with His.Tag affinity chromatography, and its bioactivity was analyzed. The results showed that GST-hHSS fusion protein was expressed both as a soluble or a inclusive body in bacterial cytosol. The soluble GST-hHSS expression reached up to 30% of the whole soluble protein of bacteria as determined by densitometry. The cleavage of GST-hHSS fusion protein with Factor Xa produced two fragments of the protein, which sized 33 and 15 kD, respectively. The molecular weight of recombinant HSS protein was identical to theoretical deduction based on the DNA sequences. The protein homology of 15 kD hHSS could be efficiently eluted out after Factor Xa cleavage. It is further indicated that the recombinant hHSS is able to proliferate hepatoma cells of BEL-7402 in the preliminary experiments. PMID- 11930237 TI - Interleukin-1beta expression and phospholipase A(2) activation after intestinal ischemia/reperfusion injury. AB - The experiments were carried out to explore the interactions between IL-1 beta gene expression, protein level and phospholipase A(2) PLA(2) inhibition after intestinal ischemia/reperfusion injury. Using a rat intestinal ischemia/reperfusion injury model, after collecting the serum, lung lavage, abdomen cavity lavage and important organ tissue samples from control, injury and PLA(2) inhibitor treated groups, IL-1 beta level was measured by radioimmunoassay, and the mRNA expression of IL-1 beta and type II PLA (2)was determined by RT-PCR. After 6 h of injury, the IL-1 beta level in serum was significantly higher than that in the control group; an increase in IL-1 beta was also observed in abdomen cavity lavage 1 or 3 h after injury. IL-1 beta was significantly increased in liver tissue after injury, but was not changed obviously in the lung, kidney and intestinal tissues. IL-1 beta in the lung lavage was significantly higher than that of control group. The mRNA expression of IL-1 beta in lung tissue was increased after injury, but type II PLA(2) mRNA expression was decreased. There were different changes in IL-1 beta level and gene expression after treatment with PLA(2) inhibitor chloroquine, cyclo-oxidase inhibitor indomethacin, or PAF receptor antagonist SR27417 respectively after injury. All these results indicate that after intestinal ischemia/reperfusion injury, the IL-1 beta level and mRNA gene expression are significantly increased, however, the relationship among IL-1 beta, PLA(2) activation and its metabolite release remains to be further elucidated. PMID- 11930238 TI - [Dexamethasone inhibits osteoblastic differentiation by down-regulation of LIM mineralization protein 1]. AB - To investigate the mechanisms of the inhibition of osteoblastic differentiation by dexamethasone (DEX), the effects of different doses of DEX on the activity of alkaline phosphatase (ALP), the synthesis of osteocalcin (OC) and the expression of collagen type I were observed in the cultured rat osteoblasts. The LIM mineralization protein-1 (LMP-1) mRNA, a positive regulator of osteoblasts, was semi-quantified by RT-PCR. The results showed that a lower dose (10(-9) mol/L) of DEX could enhance the activity of ALP, the synthesis of OC and the expression of collagen type I. However, a higher dose (10(-7) mol/L) of DEX inhibited them and down-regulated the expression of LMP-1 mRNA in osteoblasts. It is suggested that DEX stimulates osteoblast differentiation at lower dose, while at higher dose it inhibits osteoblast differentiation. The inhibitory action of DEX on osteoblast differentiation might be mediated by the down-regulation of LMP-1 mRNA. PMID- 11930239 TI - Vasodilative action of carbon monoxide on rat pulmonary artery in vitro. AB - The present study investigates the vasodilative action of carbon monoxide on rat pulmonary artery in vitro. After isolation of the pulmonary artery rings (PAR) from Wistar rats, an ACh concentration-response curve was generated; the PARs were incubated with the NOS inhibitor L-NAME (30 micromol/L, n=10) or the heme oxygenase inhibitor ZnPPIX (10 micromol/L)+L-NAME (30 micromol/L, n=10) for 30 min. After that, a second ACh concentration-response curve was elicited. Other isolated PARs were randomly divided into two groups: endothelium-intact group (n=8) and endothelium-denuded group (n=8). The effect of exogenous carbon monoxide (CO) on pulmonary arterial vessel tone was observed. The results showed that ACh induced a concentration-dependent pulmonary vasorelaxation. This relaxation disappeared after endothelium was denuded. The ACh induced relaxation was attenuated after pretreatment with 30 micromol/L L-NAME, and attenuated further after pretreatment with 10 micromol/L ZnPPIX+30 micromol/L L-NAME. Exogenous carbon monoxide relaxed pulmonary artery in both the endothelium-intact group and the endothelium-denuded group. These data suggest that ZnPPIX inhibits ACh induced endothelium-dependent pulmonary artery relaxation and that CO is an endothelium-derived relaxation factor, and exogenous CO can relax pulmonary artery. PMID- 11930242 TI - [Roles of entopeduncular nucleus in acupuncture analgesia and caudate-putamen nucleus stimulation-induced analgesia]. AB - The present study was to investigate whether entopeduncular nucleus (EP) is involved in caudate-putamen nucleus (CPu) stimulation-induced analgesia and in acupuncture analgesia. It was found that the foot-withdrawal latency elicited by radiant heat exposure was increased after electroacupuncture analgesia (EA), and the nociceptive responses of neurons in parafascicular nucleus (Pf) were inhibited after EA or after excitation of CPu neurons in normal rats, but the foot-withdrawal latency and nociceptive responses of Pf neurons were unchanged by EA or excitation of CPu in the rats with lesion of EP by local application of kainic acid. The results obtained with microinjeciton of saline instead of kainic acid into the EP were the same with those in the nonlesioned control group. The differences in the results between the lesion group and the other groups were significant ( <0.05). It is suggested that EP is involved in acupuncture analgesia and also plays an important role in caudate-putamen nucleus stimulation induced analgesia. PMID- 11930240 TI - [Effects of regulatory peptides on adhesion of eosinophil to bronchial epithelial cells]. AB - To explore the roles of regulatory peptides in the process of various anaphylactic inflammation of the airway, we observed the influence of four peptides, i.e., vasoactive intestinal peptide (VIP), epidermal growth factor (EGF), endothelin-1 (ET-1), and calcitonin gene-related peptide (CGRP), on the adhesion of eosinophil (EOS) to unstimulated and O(3)-stressed bronchial epithelial cells (BEC). From the experiments we observed that VIP and EGF decreased EOS adherence to O(3)-stressed BEC and downregulated airway inflammation; ET-1 and CGRP increased the adhesion of EOS to BEC in the inflammatory process; and CGRP aggravated O(3)-stressed reactions. The effects of ET-1 and CGRP were inhibited by W(7)and H(7). Anti-ICAM-1 antibody inhibited the adhesion of EOS to BEC, which brings to light that EOS adherence to BEC may be related to the expression of ICAM-1 of BEC. PMID- 11930241 TI - Effects of intracarotid injection of 17beta-estradiol on electrical activity of rostral ventrolateral medullary neurons in male rats. AB - The purpose of this study was to determine the effects of 17beta-estradiol (E(2)) on electrical activity of the rostral ventrolateral medulla (RVLM) neurons in rats. Male Sprague-Dawley rats were anesthetized with urethane (1.0 g/kg) and subjected to sino-aortic denervation. Blood pressure, heart rate and spontaneous discharge of RVLM neurons were recorded simultaneously. Intracarotid injection of E(2) (10 ng/kg) decreased the discharge rate from 14.46+/-0.47 to 9.73+/-0.33 spikes/s (P<0.001) in 25 out of 30 RVLM neurons, while blood pressure and heart rate showed no significant change. The inhibitory effect of E(2) on RVLM neuronal activity was rapid at the onset (within 1 min) and long-lasting (>5 min). Prior administration of antiestrogen tamoxifen (TAM) did not affect the effect of E(2). However, pretreatment with N( )-nitro-L-arginine-methyl ester (L-NAME), an inhibitor of nitric oxide (NO) synthase, significantly attenuated the inhibitory effect of E(2). In addition, NO donor 3-morpholinosydnonimine (SIN-1) potentiated the effect of E(2). These results suggest that E(2) may inhibit spontaneous electrical activity of RVLM neurons, an effect which is mediated by the activation of NOS with the resultant of NO release via nongenomic actions. PMID- 11930244 TI - [Role of melatonin in spatial learning and memory in rats and its mechanism]. AB - It has been suggested that melatonin is involved in learning and memory. In the present study, we investigated the effects of melatonin on spatial learning and memory in rats, using Morris water maze and electrophysiological methods. The results are as follows. (1) During a six-day water maze training, the mean escape latency of melatonin group in the last 4 days was 30.02+/-3.6 s, and that of control group was 18.44+/-2.7 s (P<0.01). The crossing annulus coefficient of melatonin group was 25.68+/-2.32%, and that of control group was 43.33+/-2.85% (P<0.01). (2) Microinjection of melatonin into CA1 area inhibited long-term potentiation (LTP). Sixty minutes after tetanus, the field excitory postsynaptic potentials (fEPSP) slope of group C (n=7 0.5 microliter saline) was 169.71+/-6.48 % of the baseline, and that of group M2 (n=6, 2 microgram melatonin) was 114.28+/ 1.80% of the baseline. The difference is significant (P<0.01). (3) We also investigated the effects of melatonin on LTP after administration of scopolamine. Sixty minutes after tetanus, the fEPSP slope of group SM (n=6, 2 microgram scopolamine before 2 microgram melatonin) was 113.70+/-5.55% of the baseline. It showed a significant decrease compared with group C (P<0.01). However, there was no difference between groups SM and M2 (P>0.05, i.v.). The results obtained by applying melatonin after bicuculline were different from those after scopolamine. Sixty minutes after tetanus, the fEPSP slope of group BM (n=7, 1 microgram bicuculline before 2 microgram melatonin) was 162.29+/>10.52% of the baseline. Compared with group C, there is no significant difference (P>0.05); but compared with group M2, the difference is significant (P<0.01). Our results showed that application of melatonin in rats significantly inhibited not only spatial learning and memory, but also LTP in CA1 area. Furthermore, the results indicate that the inhibition of LTP by melatonin may not be mediated by cholinergic system, but may be through the modulation of GABAergic system. PMID- 11930243 TI - [Propofol depresses c -fos expression of NOS neurons in the spinal cord of rats with inflammatory pain]. AB - In formalin pain model, the effect of propofol on Fos expression in the spinal cord was examined by means of c -fos oncogene immunohistochemistry and NADPH-d histochemistry. Fos-like immunoreactive (FLI) neurons were mainly found in the ipsilateral dorsal horn after injection of formalin, some of which were FLI/NOS double-labeled neurons. Most of the FLI or FLI/NOS double-labeled neurons were observed in the medial part of lamina and the outer lamina . Before or after injection of formain, i.p. injection of propofol significantly decreased the number of FLI and FLI/NOS double-labeled neurons in all laminae (P<0.05 or P<0.01). By single i.p. injection of propofol or normal saline, few FLI neurons were found in the spinal cord. The results suggest that the antinociceptive function of propofol is possibly involved in the depression of the NOS neurons in the spinal cord. PMID- 11930245 TI - [Expression of intercellular adhesion molecule-1 in different organs of the mice with endotoxic shock induced by lipopolysaccharide]. AB - To investigate and compare the expression of intercellular adhesion molecule-1 (ICAM-1) in different organs of the mice with endotoxic shock induced by lipopolysaccharide (LPS), protein and mRNA of ICAM-1 were measured by Western blotting and RT-PCR respectively in different organs of BALB/c mice administered intraperitoneally with 5 mg/kg LPS. The results showed that the constitutive expression of ICAM-1 protein and mRNA was the greatest in the lungs, followed by the spleen, kidney and intestine. After LPS stimulation, the upregulation of ICAM 1 was still greatest in the lungs, followed by the liver, spleen, heart, kidney and intestine. Compared with the normal mice, the expression of ICAM-1 protein in endotoxic shocked mice increased by 4.5-fold in the lungs, 3.0-fold in the kidney, 1.5-fold in the spleen; the expression in the liver and heart was negative under normal condition and changed into positive during endotoxic shock; but ICAM-1 expression in the intestine did not change significantly. The expression of ICAM-1 mRNA also increased consistently. These data highlight that LPS can up-regulate ICAM-1 protein and mRNA expression in different tissues of the mice with endotoxic shock. The difference in ICAM-1 expression among the organs may lead to different sensitivity of organ damage in endotoxic shock. This suggests that inhibition of ICAM-1 expression may be a useful principle for prevention and treatment of endotoxic shock. PMID- 11930246 TI - [Effect of angiotensin II on follicular atresia in mouse]. AB - The effect of angiotensin II (Ang II) on the follicular development was studied by using an animal model of follicular atresia induced by pregnant mare s serum gonadotropin (PMSG). The results showed that: (1) a large number of atretic follicles were found in the ovary of 24-day-old mouse after 6-day treatment of PMSG. Deoxyribonucleic acid (DNA) extracted from granulosa cells clearly showed a ladder band under agarose gel electrophoresis analysis. (2) the contents of Ang II in the ovary extremely increased with the development of follicular atresia. (3) Ang II significantly antagonized the stimulating effect of the follicle stimulating hormone (FSH) on estradiol (E(2)) generation of granulosa cells. It is suggested that Ang II may be involved in the regulation of follicular atresia in mouse. PMID- 11930247 TI - [Recombinant human interleukin-10 inhibits vascular smooth muscle cell proliferation induced by TNF-alpha]. AB - Vessel injury provokes a release in proinflammatory cytokines that influence vascular smooth muscle cell (VSMC) proliferation. The purposes of this study was to determine the effects of recombinant human interleukin-10 (rhIL-10) on rat vascular smooth muscle cell proliferation and the activity of p44/p42 mitogen activated protein kinase (MAPK) promoted by tumor necrosis factor-alpha (TNF alpha). Rat aortic VSMCs were cultured and treated with rhIL-10 or TNF-alpha respectively, and then cotreated with rhIL-10 and TNF-alpha. The proliferation of VSMCs was quantified by colormetric assay. Cell cycle analysis was performed by flow cytometry. The p44/42 MAPK activity was evaluated by the immunoblotting technique using anti-p44/42 phospho-MAPK antibody. Compared to control group, TNF alpha stimulated significantly VSMC proliferation in TNF-alpha group. rhIL-10 alone had no effect on VSMC growth, but significantly inhibited VSMC proliferation induced by TNF-alpha at a dose of 10 ng/ml. The cell number in G(0)/G(1) phase of TNF-alpha and rhIL-10 co-treatment group was higher than that of TNF- alpha group (P<0.01) by flow cytometry analysis. The p44/42 MAPK activity was significantly enhanced by TNF-alpha and the TNF-alpha effect was opposed by rhIL-10. It is suggested that rhIL-10 can inhibit TNF-alpha induced VSMC proliferation and phosphorylation of p44/42 MAPK. PMID- 11930248 TI - [Electrophysiological effects of phytoestrogen genistein on guinea pig papillary muscles]. AB - The cardiac electrophysiological effects of genistein (GST) were examined in guinea pig papillary muscle using intracellular microelectrode technique. The results obtained are as follows. (1) Duration of action potential (APD) in normal papillary muscles was decreased by GST (10 100 micromol/L) in a concentration dependent manner. (2) In partially depolarized papillary muscles, 50 micromol/L GST not only reduced APD, but also decreased the amplitude of action potential, overshoot and maximal velocity of phase 0 depolarization. (3) Pretreatment with N( )-nitro-L-arginine (L-NNA, 5 mmol/L) failed to affect the above effects of GST (50 micromol/L)on papillary muscles. (4) 17beta-estradiol (E(2), 5 micromol/L) or GST (10 micromol/L) alone did not affect action potential, while GST combined with E(2) at the same doses shortened APD significantly. All these results indicate that the effects of GST on papillary muscles are likely due to a decrease of calcium inflow which is not mediated by NO and that GST has a facilitative or synergetic action with E(2). PMID- 11930250 TI - Editorial Comment. PMID- 11930251 TI - Editorial Comment. PMID- 11930252 TI - Editorial Comment. PMID- 11930265 TI - 3rd European Meeting on Pulse Wave Analysis and Large Artery Function. PMID- 11930268 TI - Dopa-responsive dystonia -- the story so far. AB - Dopa-responsive dystonia (DRD) is an eminently treatable condition and its recognition is therefore of crucial importance. In classical cases, the disease manifests in early childhood with walking problems due to dystonia of the lower limbs. The dystonia is frequently accompanied by "parkinsonian" features such as reduced facial expression or slowing of fine finger movements. Biochemically, the disorder is typically characterized by low levels of the neurotransmitter metabolite homovanillic acid and reduced levels of neopterin and tetrahydrobiopterin (BH4) in the cerebrospinal fluid. This is due to heterozygote mutations of the GTP cyclohydrolase I gene, which is the rate-limiting enzyme in the synthesis of BH4. BH4 is an essential co-factor for tyrosine hydroxylase (TH), the rate-limiting enzyme in the synthesis of dopamine. Reduced levels of BH4 lead to the dopamine-deficit syndrome DRD because of reduced TH activity. Other genes implicated in the pathogenesis of this disorder are the TH gene itself and the parkin gene. This article summarizes all relevant aspects of DRD including recent advances in the genetics of this disorder and the widening phenotype. Particular emphasis is given to clinically relevant aspects such as diagnostic difficulties and atypical presentations in infancy and early childhood. PMID- 11930269 TI - Systemic activation of the immune system during ganciclovir treatment following intratumoral herpes simplex virus type 1 thymidine kinase gene transfer in an adolescent ependymoma patient. AB - During ganciclovir treatment of an adolescent ependymoma patient two weeks after intracranial implantation of HSVtk retroviral vector producer cells, increasing numbers of peripheral T- and B-cells were found as well as enhanced T-cell activation and elevated serum levels of interleukin 12 and soluble Fas ligand. These findings suggest the systemic activation of the immune system during ganciclovir treatment in our patient. The induction of an immune response by HSVtk/ganciclovir supports the concept of an anti-tumor vaccination effect by prodrug activating gene therapy systems and may open new promising perspectives for enhancing therapeutic efficiency by combined prodrug activating and immunological gene therapy strategies. PMID- 11930270 TI - Selective muscle involvement on magnetic resonance imaging in autosomal dominant Emery-Dreifuss muscular dystrophy. AB - OBJECTIVE: The aim of this study was to evaluate the spectrum of muscle involvement on MRI in patients with autosomal dominant Emery-Dreifuss muscular dystrophy (EDMD2) due to mutations in the lamin A/C gene and to compare it to the pattern found in other conditions with similar phenotype. PATIENTS AND METHODS: Nine patients with a diagnosis of EDMD2 had MRI scanning of their leg muscles. Seven other patients, four with the X-linked form of Emery-Dreifuss muscular dystrophy (EDMD) and three with an Emery-Dreifuss-like phenotype but no detectable mutations in either the emerin or the lamin A/C gene were also scanned as disease controls. RESULTS: All patients with EDMD2 showed a characteristic involvement of the posterior calf muscles. The medial head of the gastrocnemius was always predominantly involved while the lateral head was relatively spared. This pattern was more obvious in mildly affected patients in whom the other calf muscles were spared or only mildly involved but was also recognisable in the patients with more advanced disease. In contrast, none of the patients with the X linked EDMD or with Emery-Dreifuss-like phenotype but no mutation in either genes showed this pattern of muscle involvement. CONCLUSIONS: Our results suggest that patients with EDMD2 have a specific pattern of muscle involvement and that muscle MRI can be used, in combination with other techniques, to distinguish various genetic forms of Emery-Dreifuss muscular dystrophy. PMID- 11930271 TI - The relationship between paroxysmal kinesigenic choreoathetosis and epilepsy. AB - PURPOSE: To clarify the relationship between paroxysmal kinesigenic choreoathetosis (PKC) and epilepsy, we investigated the clinical and electroencephalographic (EEG) findings of patients with familial PKC and epilepsy, as well as sporadic cases with both PKC and epilepsy. PATIENTS AND METHODS: Patients consisted of 12 familial cases from seven families and three sporadic cases. The period of follow-up ranged from 17 months to 33 years, 7 months (average: 16 years, 8 months). During the follow-up, a total of 163 EEGs (11 EEGs per subject) were studied, including interictal and ictal EEGs. RESULTS: Transient epileptic discharges were found in ten of the 15 patients (66.7 %) during the clinical course. As for focus, centro-midtemporal and frontal spikes were most often observed. The ictal EEG of an afebrile convulsion in one patient showed a partial seizure with secondary generalization which originated from the frontal area. CONCLUSIONS: It appears that patients who suffer from both PKC and epilepsy have a functional abnormality of the cerebral cortex, particularly in the perirolandic and frontal regions. PMID- 11930272 TI - Epilepsy surgery in children with focal cortical dysplasia (FCD): results of long term seizure outcome. AB - The purpose of this study was to assess the effect of epilepsy surgery on seizure outcome in children and adolescents under 18 years with intractable epilepsy due to focal cortical dysplasia. We analysed clinical data, such as age at seizure onset, epilepsy course, localisation of focus from presurgical evaluation, MRI, tissue pathology and seizure outcome in 68 patients 6 months to 9 years after epilepsy surgery. Seizure outcome was classified according to the Engel classification. Mean age at seizure onset was 7 months, ranging from the first days of life to 7 years. All patients had medically intractable epilepsy. Localisation of the lesion was predominantly extratemporal: posterior (uni- or multilobar) 43 %, frontal without central region 26 %, multilobar involving central area 19 % and temporal in 12 %. MRI signs typically seen in cortical dysplasia (FCD) such as localised blurring of gray-white matter junction was found in 68 %, dysgyria in 62 %, thickening of the cortical ribbon in 46 % and T2 signal elongation of the subcortical white matter in 40 % of the patients' MRI. Age at surgery ranged from 5 months to 16 years; 14 patients were under 2 years when operated on. In 34 patients (6 patients under 3 years) subdural grid electrode evaluation was performed prior to surgery. Pathology revealed focal cortical dysplasia without balloon cells (type I) in 60 %, FCD of the balloon cell subtype (type II) in 40 % of the specimens. Postoperative complications were subdural hygroma in 5 and an increased motor deficit in 2 patients. Up to two years after epilepsy surgery 50 % of the children were seizure free (Engel class I), 10 % Engel class II, 33 % Engel class III and 7 % unchanged (Engel class IV). Long-term seizure outcome (> 3 years post surgery) in 32 patients showed similar results (class I 50 %, class II 19 %, class III 28 %, class IV 3 %). Complete resection of the dysplastic lesion was significantly correlated with favorable seizure outcome, whereas seizure outcome was not significantly different in patients with mild (type I) or balloon cell (type II) FCD. Children operated after 6 years of age had no better outcome than children operated in infancy or at preschool age. Epilepsy surgery resulted in good (class I and II) seizure control in 60 % of children with intractable epilepsy due to focal cortical dysplasia. PMID- 11930273 TI - Congenital disorder of glycosylation IId (CDG-IId) -- a new entity: clinical presentation with Dandy-Walker malformation and myopathy. AB - A 1.5-year-old boy with macrocephaly due to a Dandy-Walker malformation presented with progressive hydrocephalus, extensive muscular hypotonia, transient cholestatic syndrome, extensive coagulation abnormalities and elevated creatine kinase indicating myopathy. Diagnostic work-up indicated a congenital disorder of glycosylation (CDG, formerly carbohydrate deficient glycoprotein syndrome). The serum transferrin pattern obtained by automated isoelectric focusing (IEF) showed an hitherto unreported pattern with strongly elevated tri-, di-, mono- and asialotransferrin bands, increasing in this order together with markedly decreased tetrasialotransferrin. Investigation of two additional glycoproteins, alpha(1)-antitrypsin and alpha(1)-antichymotrypsin, confirmed a generalised defect of glycosylation. All known glycosylation defects could be ruled out by enzymatic analyses in either leukocytes or fibroblasts or by the results obtained by IEF. SDS-electrophoresis demonstrated a marked difference in the molecular weight of transferrin, suggesting the lack of parts or of all oligosaccharide chains. The defect could be delineated to a deficiency of beta-1,4 galactosyltransferase (E.C.2.4.1.38) due to a homozygous insertion (1031 - 1032 insC). Details of the biochemical and molecular findings will be described elsewhere. PMID- 11930275 TI - Large cerebral vessel occlusive disease in Lyme neuroborreliosis. AB - We report on a 12-year-old, previously healthy girl with an acute hemiparesis as the predominant clinical manifestation of Lyme neuroborreliosis (LNB). The diagnosis of LNB was based on cerebrospinal fluid (CSF) studies, laboratory findings and the clinical course whereas the patient's history and the lack of characteristic skin lesions obscured the diagnosis in the beginning. After four weeks of antibiotic and physiotherapeutic treatment, the hemiparetic symptoms had completely resolved. Although evidence of vasculitic and perivascular inflammation in LNB has been described in the literature, large cerebral vessel occlusive disease represents a rare finding. Appropriate treatment strategies can lead to good clinical rehabilitation, as shown in this case, making the timely diagnosis a crucial issue. We conclude that LNB should be considered in every stroke-like episode of unknown origin in children, even in the absence of a history of a tick bite or typical skin lesions. PMID- 11930274 TI - MECP2 mutation in a boy with severe neonatal encephalopathy: clinical, neuropathological and molecular findings. AB - We describe the clinical and neuropathological presentation of a male with an MECP2 mutation whose sister has Rett syndrome (RS). He presented with severe neonatal encephalopathy and died at the age of 13 months. Mutation analysis of the MECP2 gene demonstrated a 488 - 489 del mutation in his and his sister's copies of the gene. Post mortem examination revealed bilateral polymicrogyria in the perisylvian region. This malformation was visibly more severe than previously described in females with RS and another male with an MECP2 mutation. As bilateral polymicrogyria was described in congenital perisylvian syndrome, the presented patient could be regarded as having suffered from a severe form of this syndrome. We conclude that MECP2 screening should be considered in males with severe neonatal encephalopathy and in males and females with a bilateral polymicrogyria syndrome. PMID- 11930276 TI - Breath holding spells in a 3-day-old neonate: an unusual early presentation in a family with a history of breath holding spells. AB - Breath holding spells are common in childhood. They peak about 2 years of age and abate by 5 years of age; they are rare before 6 months of age. We report a case of cyanotic breath holding spells starting at the age of 3 days. There was a family history of breath holding spells including a three-year-old brother, patient's father, his sister, and the paternal great grandfather. Pharmacological intervention with iron replacement therapy and piracetam was not successful. We want to draw attention to the fact that breath holding spells should be included in the differential diagnosis of neonatal cyanotic spells. The course of these neonatal breath holding spells is thought to be "benign", but nevertheless, severe breath holding spells can pose problems for parents and clinicians. PMID- 11930277 TI - Resistance to rocuronium in a child with Schwartz-Jampel syndrome type 1 B. AB - In Schwartz-Jampel syndrome micrognathia and jaw muscle rigidity may result in difficult or impossible tracheal intubation. Since the dose-response relationship to muscle relaxants is unknown in this rare disease we assessed by mechanomyography the neuromuscular response to the rocuronium in a two-year-old child with Schwartz-Jampel syndrome (SJS) Type 1 B. Rocuronium's dose-response curve was markedly shifted (3.5-fold dose) to the right when compared to healthy children and intubation conditions were improved. This resistance to NDMR may result from a lower acetylcholine degradation rate suggested as being the consequence of mutation of the gene encoding perlecan (HSPG2) in SJS. Thus, considerably higher doses of NDMR than usual may be required for facilitation of tracheal intubation in patients with SJS. Since evidence for genetic heterogeneity of SJS exists we also recommend incremental doses of a rapidly acting NDMR with continuous monitoring of neuromuscular function so as to assess the optimum relaxant dose. PMID- 11930278 TI - D-glyceric aciduria in a six-month-old boy presenting with West syndrome and autistic behaviour. AB - D-Glyceric aciduria is a disease with a very heterogeneous group of symptoms, with D-glyceric acid excretion as the chief common characteristic. Findings described in previous patients include progressive neurological impairment, hypotonia, seizures, failure to thrive and metabolic acidosis. However, there are also asymptomatic patients with mild neurological impairment. A six-month-old boy was admitted to our clinic with the complaints of dullness to his environment, seizures and autistic behaviour. EEG revealed multifocal generalized epileptic activity in a hypsarrhythmia pattern. Organic acid analysis (GC-MS) in urine revealed increased glyceric acid excretion. Analysis of the optical form of glyceric acid by a polarimetric method supported the diagnosis of D-glyceric aciduria. MRI showed white matter lesions with cerebral atrophy, particularly in the frontotemporal regions, and reversible abnormalities in the mesencephalon, thalami and globus pallidium resolving after fructose restriction in the diet. To our knowledge, this is the first case report of a patient with D-glyceric aciduria who presented with West syndrome and autistic behaviour in whom serial MRI findings are also defined. PMID- 11930279 TI - Undetectable levels of CSF hypocretin-1 (orexin-A) in two prepubertal boys with narcolepsy. AB - We report on two prepubertal narcoleptic boys with undetectable levels of hypocretin-1 (orexin-A) in their cerebrospinal fluid (CSF). The disease onset times were 6 and 8 years, and CSF was collected 8 and 20 months after the onset, respectively. The initial symptoms were excessive daytime sleepiness, cataplexy and disrupted nocturnal sleep. Both subjects are DRB1*1501 and DQB1*0602 positive. The measurement of CSF hypocretin-1 is valuable for the decisive diagnosis of narcolepsy and for selecting the type of treatment in prepubertal children. Our results suggest that a significant degree of hypocretin deficiency is already present at the disease onset. PMID- 11930280 TI - Alice in Wonderland syndrome: a clinical presentation of frontal lobe epilepsy. PMID- 11930282 TI - [For what we need clinical nutrition]. AB - Clinical nutrition covers a wide range of specialties of medicine. There is a need for better nutritional support in prevention of chronic diseases, investigations to assess malnutrition, dietetics including artificial nutrition and rehabilitation as well as supportive care for the severely and terminally ill patients. Clinical nutrition follows the principles of evidence-based medicine. In future the training of doctors in human nutrition has to be further improved. Since nothing works well in isolation an integration of clinical nutrition into gastroenterology is recommended. PMID- 11930283 TI - [Tube feeding - who, how and when]. AB - Summary. The concept of enteral nutrition is well established for a long time. During the past twenty years different percutaneous tube feeding techniques have been established. The most popular method is the percutaneous endoscopic gastrostomy (PEG) which is applied as a pull-through technique in Germany. In some patients this approach is not successful and alternative techniques must be used (e. g. introducer method). In some patients nasal or percutaneous feeding tube have to be placed directly into the intestine. There are different systems available for this approach which have to be clearly indicated. As a second step after initiation of enteral nutrition therapy quality of life can be improved by implanting secondary systems (e. g. button gastrostomy).A standardized technique for inserting tubes is essential to have a successful long-term outcome in enteral nutrition and care after has to be integrated into the regimen. The aim of this article is to demonstrate different enteral nutrition tube techniques their indication, contraindication and long-term follow-up. PMID- 11930284 TI - [Metabolic syndroma long term results]. AB - Several studies in obesity verify more effectiveness in weight-loss if diet and exercise were combined with behavioral modification. The main problem with all obesity treatments is a return to baseline weight after treatment ends. We have some evidence, that long-term behavioral methods may be successful to induce long term weight-loss maintenance. Strategies for improving weight-loss maintenance in rehabilitation are discussed PMID- 11930285 TI - [Obesity treatment in specialized rehabilitation centers - the "Reha-Team" and process quality]. AB - Obesity is a chronic disease with high economic relevance in particular for mutual pension funds. Evidence based guidelines describe strategies for weight reduction and maintenance. Successfull therapy requires trained staff of various professions with special skills in patient education and behaviour therapy enabling patients to make informed decisions. Using methods of quality management the process of obesity management can be optimized by training programmes as well as in-house guidelines for new obesity team members and by continued medical education in metabolic diseases and in patient communication. PMID- 11930286 TI - [Health education for chronic liver and pancreatic diseases: Discussion groups "chronic liver disease" (CL), "chronic pancreatic disease" (CP)]. AB - The article deals with our experience in the educational programs for patients with chronic liver diseases and chronic pancreatic diseases.The structure of our educational programs is modular, the main event is the discussion group "Chronic Liver Diseases" (CL) (2 meetings are headed by physicians, 1 by psychologists, duration in all 3,6 hours) respectively the discussion group "Chronic Pancreatic Diseases" (CP) (2 meetings headed by physicians, 1 by nutrition consultants, duration in all 2,6 hours). As needed, additional modules can be added, for example specific seminars in case of alcohol abuse or educational programs for diabetics. The average number of participants is between 7 - 15 persons in the discussion group CL, respectively 11 - 20 in the discussion group CP. An overhead projector is used to address the various topics, the patients are encouraged to actively participate in the discussion. The contents can be found in a structured curriculum and focus on anatomical basics, diagnostic and therapeutic aspects, physical fitness and performance in professional life. PMID- 11930287 TI - [Results of a structured hypertension teaching and treatment programme]. AB - The benefit of good blood pressure control in patients with arterial hypertension has been repeatedly demonstrated. In high risk patients, a good blood pressure control is one of the main prerequisites for the reduction in morbidity and mortality. However, as shown in population-based studies, the quality of blood pressure control both in patients with essential hypertension and in hypertensive diabetic patients is still unsatisfactory. Only a minority of patients achieve target blood pressure values below 140/90 mm Hg. This situation has not changed within the recent years, although the prescriptions of antihypertensive drugs have continuously increased. This paper describes a structured hypertension treatment and teaching programme (HTTP) which was developed and evaluated during the last 16 years and which aims at intensification of antihypertensive therapy by active involvement of the patient in his own treatment. This programme primarily focuses on the improvement of the patients' long-term treatment compliance by means of information about non-pharmacological and pharmacological therapies and instructions to regular blood pressure self measurements. The efficacy and feasibility of the HTTP has been evaluated in several studies which unanimously show considerable benefits not only concerning blood pressure control but, most importantly, regarding reduction in the incidence and progression of hypertension associated diseases as cerebro- and cardiovascular events and diabetic nephropathy. Therefore, it seems essential to implement the HTTP in the German health care system in order to finally achieve the desired quality of care for patients with arterial hypertension. PMID- 11930288 TI - [Psychologic concepts for correction of hypoglycemia-unawareness and reduction of hypoglycemia-anxiety]. AB - summary. Patients with Type-1- Diabetes have around 3 - 5 Hypoglycemias weekly, if they adjust to a nearly normal blood glucose level. Severe Hypoglycemia are rare, but often ar ed bei the patients, especially if they have Hypo-unawareness. This anxiety can provoke a behaviour of avoidness, so these people try to avoid even mild Hypoglycemia. They attitude themselves to high niveau of blood glucose. So they have a higher risk to get complications or they are living with an over estimated behaviour of safety. Hypo-unawareness and Hypo-anxiety have an urgent need of psychotherapy. PMID- 11930289 TI - [Diabetes and assessment of working capability--neuropathy and autonomic neuropathy]. AB - Up to now no obligatory guidelines for the assessment of working capability exist for patients with diabetes and peripheral and/or autonomic neuropathy. Precise investigations of clinical symptoms of an existing polyneuropathy are preconditions for this assessment. For the evaluation of the working capability a detailed analysis of working conditions and working place is necessary. Special attention is needed with respect to the prophylaxis of the diabetic foot. Cardiac autonomic neuropathy as well as the autonomic neuropathy of the gastro-intestinal tract are of importance for the assessment of working capability. In practice inadequate diagnostic procedures occur up to now and assessment of working capability is mostly not included into these diagnostic procedures. The processing of corresponding recommendations should be a common task of diabetologists and job physicians. PMID- 11930290 TI - [Driving capacity of diabetics against the background of legal principles]. AB - Within the conversion of the 2nd Council Directive on Driving Licences of the Council of the European Communities (91/439/EEC) into national law, changes became necessary to the Road Traffic Act (StVG) and a new Driving Licence Ordinance (FeV) was created which has been in force since 01.01.1999 (Federal Law Gazette, 1998, Part I No. 55, 26. August 1998). Both in Annex III of the EC Directive and in Annex 4 of the Driving Licence Ordinance, statements are made with regard to the driving capability of diabetics. Besides this legal basis for evaluating capacity to drive a motor vehicle, the appraisal guidelines (Begutachtungs-Leitlinien zur Kraftfahrereignung) established in commission of the German Federal Ministry of Transport, Building and Housing are to be understood as a set of rules which are outside the framework of actual legal statute as such and which are intended to support assessors in making decisions in individual cases. When considering whether an appraisal of driving capacity has to take place, in particular a difference is made between the different classes of vehicle. For motor cycles and passenger cars (Classes A and B), it will also be the case in future that a medical examination will only be ordered if there is a specific reason. Apart from this, only a sight test is necessary, as has been the practice up to now. In the case of lorries and buses (Classes C and D) and the licence for transportation of passengers (on a commercial basis) there will be an initial appraisal, as has also been the case up to now, and there will also be repeat appraisals. This is new for Class C, where up to the present there has only been an initial appraisal. The basic principle which applies is that the driving licence authority has the duty to instigate appraisal of the driving capacity of an applicant for a driving licence or of the holder of a driving licence if there is justified doubt about the driving capacity of the person concerned. PMID- 11930291 TI - [Evaluation of osteoporosis]. AB - Physical examination, bone densitometry and x-ray are the mainstay in the evaluation of osteoporosis. A limited number of simple blood tests are sufficient to discriminate most secondary forms of osteoporosis and to exclude osteomalacia. Bone-specific biochemical markers play an increasing role in monitoring osteoporosis therapy. They may also play a role in treatment decision in the coming years. Bone biopsies are helpful if findings are unclear or inconsistent. The present review gives a summary of the opportunities and limitations of the above diagnostic procedures. PMID- 11930293 TI - [Role of physical activity for the prevention and rehabilitation of osteoporosis]. AB - The therapeutic exercises pursue preventive as well as rehabilitative aims within the osteoporosis therapy. The objective depends on whether an osteoporosis already occurred and it depends also on the respective stage of the osteoporosis. Aims and main points from the physiotherapy and sports therapy are: construction of the bone substance; receipt of the bone substance; pain alleviation; fall prophylaxis. To reach an optimal bone mineral content in recent years activities are necessary on which own body weight must be carried. In the later adult age the same activities in addition are necessary to reduce the natural reduction rate of bone mineral content to a minimum. With beginning or already occurred osteoporosis must those activities be operated with caution and restraint which have to do with abrupt interceptions of the body weight.The combination of medication and physical activities shows better therapy results as the respective measures alone. An important therapeutic aspect with older humans exists in the reduction of the fall risk and thus the fracture risk. In addition a muscle training - as for example in the medical training therapy - can raise the pain threshold, so that the pain sensitivity of the patients is improved. PMID- 11930292 TI - [Osteoporosis - Evidence based therapy]. AB - Osteoporosis therapy has been controversially discussed in the past. In the meantime, several therapeutic options to prevent fractures are available for this disease. With respect to proven fracture benefit, however, the quality of evidence from randomised and controlled clinical trials varies substantially among therapies. From systematic research the best external evidence is available for a supplementation with calcium and vitamin D and a therapy with the bisphosphonates alendronate or risedronate, as well as the SERM raloxifene. For other therapeutic agents like fluorides, vitamin D metabolites, calcitonin and etidronate the quality of evidence is much lower. So far, there is no evidence for other pharmaceutical therapies. Hip protectors are effective in the prevention of hip fractures. PMID- 11930294 TI - [Indications for chemotherapy in cancers of the esophagus, stomach and pancreas]. AB - During the last years the chemotherapy in osophageal, stomach and pancreatic cancer demonstrated some success. Radiochemotherapy for esophageal cancer is indicated as neoadjuvant therapy before surgery in locally advanced cancer or in patients with other diseases, which do not allow surgery. In stomach cancer patient there is a clear indication for chemotherapy in metastatic disease and within clinical trials as neoadjuvant chemotherapy in locally advanced cancer. In pancreatic cancer patient the chemotherapy shows less success comparing to other gastrointestinal cancer; it is part of the palliative concept with other therapeutic strategies. PMID- 11930295 TI - [Anorectic syndrome]. AB - The "anorectic syndrome" of patients with malignant diseases is a symptom complex consisting of loss of appetite, aversion against food and disturbances of taste and smell. Anorexia is defined as a decrease of nourishment below 80 % of the usual food intake. Occurrence of anorexia depends on the type and location of the tumor and on tumor stage. About 40 % of tumor patients suffer from anorexia already at the time of diagnosis. The highest prevalence is seen in advanced tumor stages the etiology of anorexia is multifactorial. The causes mentioned include food aversion, disturbances of taste, gastrointestinal disorders, pain, treatment sequelas, consequences of metabolic disturbances, effects and interactions of cytokines, neuropeptides and neurotransmitters, reduced exercise as well as psychological factors and conditioned aversions. Treatment options of anorexia consist of nutritional measures like "preference guided nutrition" considering the patients individual aversions and preferences. Drugs recommended for treatment are procinetic agents, corticosteroids, gestagens or cannabinoids PMID- 11930296 TI - [Handling of cytostatics and patients care in chemotherapy]. PMID- 11930297 TI - [Nursing care and companionship of patients in cytostatic therapy in a rehabilitation-clinic]. PMID- 11930298 TI - [Rehabilitation after surgery for gastrointestinal malignant tumors]. AB - In Germany every year about 100,000 people develop a malignant tumor of the gastrointestinal tract. Only few of them are included in a structured rehabilitation program after surgery. The aim of such a rehabilitation program is to minimize the sequels of gastrointestinal tumor disease and its treatment and influence on the daily life of patients. Malnutrition is a common symptom of tumor manifestation, especially in the gastrointestinal tract. It is important to establish the diagnosis of malnutrition and to treat such patients with nutritional support.Fatigue and impairment of physical performance are serious problems of cancer patients. Particularly in the recovery phase immediately after treatment, low physical performance imposes limitations on basic daily activities. An active aerobic training program can improve the physical performance and fitness in a short time. Deficits and symptoms depend on tumor stage and localisation. A specialised team is necessary to treat all the possible symptoms and complications after cancer surgery. Patients have to be informed precisely about structural and functional deficits due to tumor operation. Only the well informed patient will comply to the suggested treatment. PMID- 11930299 TI - [Chronic inflammatory bowel disease - new therapies: tumor necrosis factor antibodies and cytokines]. AB - The chimeric antibody directed against tumor necrosis factor- alpha (TNF alpha) Infliximab (Remicade) is effective in the therapy of Crohn's disease. This is rather surprising since it was unlikely that single factor should be effective, considering the complexity and redundance of the mucosal cytokine system. Because of safety reasons and high cost the indication should be restrictive. PMID- 11930300 TI - [Extra-intestinal Manifestations of chronic inflammatory Bowel diseases]. AB - Approximately one third of all patients with Crohn's disease and ulcerative colitis suffer from extra-intestinal manifestations of their inflammatory bowel disease. Most commonly those symptoms occur simultaneously with the CED symptoms; they can, however, either precede them or appear later on in the course of the disease. The most frequent extra-intestinal symptoms are arthralgias of peripheral joints and spine, which are usually bland and self-limiting, while ankylosing spondylitis and erosive arthritides are rare. Skin lesion and eye affections can also parallel the bowel condition, but occasionally they precede intestinal manifestations and can be a first diagnostic clue. In addition, different extra-intestinal symptoms tend to simultaneously co-occur more frequently in some patients with CED, while others are not affected at all. Immunogenetic parameters play a role for the manifestations of the pathologic immune response both in the gut and in the musculo-skeletal systems, as indicated by associations with MHC class I alleles. Enteropathic microorganisms are also thought to be involved in the pathogenetic mechanisms. PMID- 11930301 TI - [Place of psychotherapy in the treatment of chronic inflamatory bowel disease]. PMID- 11930302 TI - [Information programs for patients during gastroenterological rehabilitation]. AB - Patient education especially groups for patients with inflammatory bowel disease belong to the typical tasks of Reha-medicine. In the Reha-Clinic "Ob der Tauber", Bad Mergentheim these patients are informed in special Colitis/Crohn-groups for eleven years now. Normally approximately ten patients with IBD are treated in our house at the same time. These patients participate in a nearly closed group in which they are taught in three weeks blocks. Three hours a week the patients are informed by doctors, one hour a week a diet assistant teaches the participants. The psychotherapeutic group takes place regularly once a week. The aim of the patient information or health education is to reach a mostly complete information of the patients about inflammatory bowel disease, that means to inform them about all diagnostic and therapeutic possibilities. Another important purpose of team work is the support of dynamic group processes like reduction of anxiety and the influence on the patients for developing an active positive attitude towards their disease. The participating therapist (doctors, psychotherapist, diet assistant) have created a structured concept which was modified during the last years. Resulting from the interactive work between therapist and patients this concept is improved continuously. This development however is delayed by the shortcomming of bad personal capacities and the lack of possibilities for the therapist to improve their pedagogic and psychotherapeutic abilities. At the moment their education is based on autodidactic methods and empathy. For a standardization of the program including all hospitals the extended curriculum of the GRVS is useful. An evaluation concerning the therapeutic benefit of patient information has not been done yet. PMID- 11930303 TI - [Vocational assessment of diarrhea, incontinence and stoma]. AB - The assessment of capability of work consist of measuring the patient subjective willingness to perform and his efficiency which should be objectively examined by functional diagnostic tools. On the one hand, there is the consideration of the patients psycho-mental burdens and on the other hand has to be defined the extent of organic functional restrictions. The latter is difficult to assess because diseased organs of the gastrointestinal tract are not directly involved in the efficiency of work. In chronic diarrhea purely watery diarrhea causes mobility restrictions at the place of work. In case of chronic malabsortive diarrhea, psycho-mental and physical deficits of efficiency have to be taken into account. The latter however may be almost restored by adequate substitution of macro and micro nutrients. For the clinical assessment of the severity of incontinence and ist psychosocial effects structured history sheets and working up of clinical findings and specifics function tests are necessary. The so defined degree of stool incontinence determines the restrictions of vocational efficiency. Generally persons with stoma are nearly not disabled and capable of gainful employment. This also pertains to professions with specific hygienic requirements, such as a production and distribution of foods. Persons with well applied and provided stomata should avoid severe physical burdens, unsuitable postures and periodical forms of labour like piece-work. PMID- 11930304 TI - [Shiftwork]. AB - First the most important problems of shiftworkers are discussed, i. e. living against the "inner clock", disturbances of sleep and appetite, gastrointestinal complaints, diseases, disturbances of the social life, faults and accidents. Thereafter a list of potenzial measures to reduce the problems of shiftworkers is presented and one measure -the design of shift systems according to new ergonomic recommendations- is explained. PMID- 11930305 TI - [Current aspects of hepatotoxicity in occupational medicine]. AB - A large number of chemical agents encountered occupationally are known as potential hepatotoxins. The practical relevance of these results should be considered critically for the individual case and under usual work layout. In this paper the current state of knowledge for hepatotoxicity in occupational medicine is described. Different diagnostical methods and criteria for suspected chemical-induced liver injury are shown. The paper focuses on the potential hepatotoxic effects of chronic exposures to solvents, since these substances are used in a large variety of modern places of work. On the basis of field studies in occupationally exposed workers the potential hepatotoxical effects of mixtures of solvents are discussed. Recommendations for medical surveillance places of work are proposed. PMID- 11930306 TI - [Biomaterial regulation (biostoffverordnung) - aspects of the practical conversion]. AB - German Biomaterial regulation (Biostoffverordnung) applies for activities with biological materials (BM). The purpose of this regulation is protection of employees working with BM. BM are micro organisms, inclusively genetically changed micro organisms, cell cultures and human pathogene endoparasits, which cause infections, sensitizing or toxic effects on humans. This regulation describes the necessary organizational, safety-relevant, personally and hygiene measures for the prevention of the diseases and damages caused by BA. PMID- 11930307 TI - [Ambulant rehabilitation of patients with gastrointestinal and metabolic diseases]. AB - For the following reasons it has, despite plausible different opinions, seldom been possible realize ambulant rehabilitation offers:For historical reasons gut rehabilitation centers lie outside of conurbations. A model of gastrointestinal oncologic rehabilitation supports developing an entirely new concept for ambulant treatments, instead of merely copying the presently in-patient rehabilitation therapy. In the exemplary thoughts of the GRVS, which were already presented five years ago, quantitative factors have not been taken into consideration satisfactorily. Considering this the following three diagnosis groups become relevant - diabetes m. - obesity - gastrointestinal cancer. The needed ambulant rehabilitation concepts are to be connected to the long-term ambulant treatment, available close to the patient's home city. PMID- 11930308 TI - Risk of influenza A (H5N1) infection among poultry workers, Hong Kong, 1997-1998. AB - In 1997, outbreaks of highly pathogenic influenza A (H5N1) among poultry coincided with 18 documented human cases of H5N1 illness. Although exposure to live poultry was associated with human illness, no cases were documented among poultry workers (PWs). To evaluate the potential for avian-to-human transmission of H5N1, a cohort study was conducted among 293 Hong Kong government workers (GWs) who participated in a poultry culling operation and among 1525 PWs. Paired serum samples collected from GWs and single serum samples collected from PWs were considered to be anti-H5 antibody positive if they were positive by both microneutralization and Western blot testing. Among GWs, 3% were seropositive, and 1 seroconversion was documented. Among PWs, approximately 10% had anti-H5 antibody. More-intensive poultry exposure, such as butchering and exposure to ill poultry, was associated with having anti-H5 antibody. These findings suggest an increased risk for avian influenza infection from occupational exposure. PMID- 11930309 TI - Illness severity, viral shedding, and antibody responses in infants hospitalized with bronchiolitis caused by respiratory syncytial virus. AB - The relationships between host factors, viral shedding, illness severity, and antibody response in respiratory syncytial virus (RSV)-induced bronchiolitis are poorly defined. These relationships were prospectively evaluated in 77 infants hospitalized with RSV bronchiolitis in multicenter, double-blind, placebo controlled trials of RSV immunoglobulin therapy. Severity of illness was influenced by age and host risk factors but was not influenced by RSV neutralizing antibody titer or by the amount of virus in nasal secretions at enrollment. Virus recovery in nasal secretions was variable but was highest at enrollment. Viral shedding was not influenced by primary diagnosis, antibody titer, age, or duration of acute respiratory illness before enrollment. In intubated patients, the amounts of virus recovered in nasal secretions and endotracheal aspirates were highly correlated. A serum neutralizing antibody response was seen in 64% of subjects who received placebo. The response was not influenced by age, primary diagnosis, amount of virus recovered, or severity of illness but was suppressed by preexisting antibody. PMID- 11930310 TI - Seroprevalence and coinfection with herpes simplex virus type 1 and type 2 in the United States, 1988-1994. AB - Seroprevalence of and coinfection with herpes simplex virus type 1 (HSV-1) and type 2 (HSV-2) in the United States were analyzed by use of data from a nationally representative survey (National Health and Nutrition Examination Survey III, 1988-1994). Evidence was explored for possible protection by prior HSV-1 infection against infection and clinical disease with HSV-2. Overall, 27.1% of persons aged > or =12 years were seronegative for HSV-1 and HSV-2; 51.0% were seropositive for HSV-1 only, 5.3% for HSV-2 only, and 16.6% for both HSV-1 and HSV-2. The seroprevalence of HSV-2 was higher in persons with HSV-1 antibody. Approximately 76% of persons who had HSV-2 antibody also had HSV-1 antibody. Persons seropositive for HSV-2 only reported a history of genital herpes more frequently (16.2%) than persons seropositive for both HSV-1 and HSV-2 (5.9%). The seroprevalence of HSV-1 and age at infection may influence the epidemiology of clinical genital herpes, even if prior HSV-1 infection does not prevent HSV-2 infection. PMID- 11930311 TI - Comparative analysis of CD8+ T cell responses against human cytomegalovirus proteins pp65 and immediate early 1 shows similarities in precursor frequency, oligoclonality, and phenotype. AB - CD8+ T cells are key effectors of the immune response against human cytomegalovirus (HCMV). A number of HCMV-derived CD8+ T cell epitopes are known. Using epitope prediction and subsequent testing for interferon-gamma responses by the ELISPOT assay, we identified an optimal human leukocyte antigen (HLA)-A*0201 restricted CD8+ T cell epitope derived from the major immediate early 1 (IE-1) gene product. As many as one-third of HLA-A*0201-positive, HCMV-seropositive donors make responses to this peptide (residues 316-324 [VLEETSVML]), which can exceed responses against a published immunodominant pp65 epitope (residues 495 503 [NLVPMVATV]). Major histocompatibility complex peptide tetramer staining facilitated detailed phenotypic analyses and revealed populations that resemble terminally differentiated effector cells (CD57+ and CD28-), with considerable restriction in T cell receptor beta-chain variable region use. The results confirm that, although pp65 is a major target for CD8+ T cells, the IE-1 protein may itself stimulate comparable responses in some persons. PMID- 11930312 TI - Suppression of human immunodeficiency virus replication during acute measles. AB - To determine the effect of measles virus coinfection on plasma human immunodeficiency virus (HIV) RNA levels, a prospective study of hospitalized children with measles was conducted between January 1998 and October 2000 in Lusaka, Zambia. Plasma HIV RNA levels were measured during acute measles and 1 month after hospital discharge. The median plasma HIV RNA level in 33 children with measles who were followed longitudinally was 5339 copies/mL at study entry, 60,121 copies/mL at hospital discharge, and 387,148 copies/mL at 1-month follow up. The median plasma HIV RNA level in children without acute illness was 228,454 copies/mL. Plasma levels of immune activation markers were elevated during the period of reduced plasma HIV RNA. Plasma levels of several potential HIV suppressive factors also were elevated during acute measles. HIV replication is transiently suppressed during acute measles at a time of intense immune activation. PMID- 11930313 TI - Elevated mucosal addressin cell adhesion molecule-1 expression in acquired immunodeficiency syndrome is maintained during antiretroviral therapy by intestinal pathogens and coincides with increased duodenal CD4 T cell densities. AB - Reduced intestinal CD4 T cell numbers and gastrointestinal disease are common features of acquired immunodeficiency syndrome (AIDS). Duodenal lymphocyte densities and mucosal addressin cell adhesion molecule (MAdCAM)-1 expression were analyzed in patients with AIDS after highly active antiretroviral therapy (HAART). Compared with human immunodeficiency virus (HIV)-seronegative individuals, HAART-naive patients with AIDS displayed reduced duodenal CD4 T cell densities. After HAART, AIDS patients with opportunistic intestinal pathogens displayed greater increases in duodenal lamina propria (LP) CD4 T cell densities than patients without such infections. Duodenal MAdCAM-1 expression was elevated in all HAART-naive patients with AIDS but remained elevated only in the intestinal pathogen group after HAART. The data suggest that, in HIV-1 infection, lymphocyte migration to the intestine may be promoted by increased MAdCAM-1 expression. After HAART, opportunistic intestinal pathogens maintain elevated MAdCAM-1 expression, which results in prominent increases in LP CD4 T cell densities in the absence of HIV-mediated CD4 T cell destruction. PMID- 11930314 TI - Is human immunodeficiency virus RNA load composed of neutralized immune complexes? AB - During acute human immunodeficiency virus (HIV) infection, both virus load (HIV RNA) and infectivity are high (10(3)-10(7) RNA copies/mL or TCID(50)/mL) until antibody is produced, which may reduce the HIV infectivity. In HIV carriers, the HIV RNA load is elevated (10(3)-10(5) copies/mL), but infectivity is low (10(0) 10(2) TCID(50)/mL). The low infectivity in carriers could be due to neutralization by antibody in serum, resulting in immune complexes (ICs). We demonstrated that ICs in plasma, prepared with protein A beads, contained HIV RNA (80%-100%) in association with immunoglobulin G (IgG). In comparison, ICs from patients with acute HIV infection and little or no antibody contained virtually no HIV RNA. Moreover, ICs prepared by ultrafiltration contained IgG and specifically and irreversibly neutralized HIV, which indicates that the ICs contained neutralizing antibody. These findings indicate that the HIV RNA in the plasma of carriers is frequently composed of antibody-neutralized HIV as ICs. PMID- 11930315 TI - Response to treatment and disease progression linked to CD4+ T cell surface CC chemokine receptor 5 density in human immunodeficiency virus type 1 vertical infection. AB - The factors governing interindividual variability in disease progression among children vertically infected with human immunodeficiency virus type 1 (HIV-1) remain unclear. Because it has recently been shown in infected adults that the density of CC chemokine receptor 5 (CCR5) molecules at the surface of nonactivated (human leukocyte antigen [HLA]-DR(-)) CD4+ T cells correlates with disease progression, the same correlation was sought in children. HLA-DR(-)CD4+ T cell surface CCR5 density was constant over time and correlated with the bioclinical stage and with the CD4 cell slope observed before antiretroviral treatment. In addition, CCR5 density was negatively correlated with the intensity of the decrease in viremia during antiretroviral therapy and was positively correlated with CD4 cell slope since birth. These results are compatible with the hypothesis that CCR5 density is a key factor governing disease progression in pediatric HIV-1 infection and, thereby, an indicator of prognosis. Moreover, they suggest that therapies aimed at reducing CCR5 accessibility should slow down HIV disease evolution in children. PMID- 11930316 TI - Virologic and immunologic response to regimens containing nevirapine or efavirenz in combination with 2 nucleoside analogues in the Italian Cohort Naive Antiretrovirals (I.Co.N.A.) study. AB - This nonrandomized study compared the virologic and immunologic responses to potent regimens containing either efavirenz or nevirapine after considering potential systematic differences between patients receiving these drugs. Virologic failure was defined as the first of 2 consecutive measurements of virus load >500 human immunodeficiency virus RNA copies/mL. Of the 694 patients included in the analysis, 460 (66.3%) started nevirapine and 234 (33.7%) started efavirenz. The adjusted relative hazard of virologic failure for patients who started nevirapine, compared with those who started efavirenz, was 2.08 (95% confidence interval, 1.37-3.15; P=.0006). In addition, patients receiving efavirenz tended to recover 5 CD4 cells/microL more per quarter (P=.05). Although comparisons of drug efficacy in nonrandomized studies should be viewed with caution, no results from randomized controlled comparisons of these drugs are thought to be available. The findings of this study are in agreement with those of other observational studies. PMID- 11930317 TI - Selection of zidovudine resistance mutations and escape of human immunodeficiency virus type 1 from antiretroviral pressure in stavudine-treated pediatric patients. AB - The relationship between clinical changes in stavudine activity and stavudine resistance was investigated in 16 human immunodeficiency virus (HIV)-infected children who received stavudine monotherapy for 18 months. Seven patients responded well to stavudine therapy, 3 experienced transient reductions in virus load, and all others had no detectable virologic response. In both the responders and nonresponders, no changes in stavudine susceptibility or specific baseline/emergent mutations in reverse transcriptase were observed. Only posttherapy HIV isolates from transient responders had elevated IC(50) values for stavudine. In 2 of the 3 transient responders, substitutions at codons 41, 210, and 215 were selected. The significance of these mutations was confirmed in viral competition experiments, site-directed mutagenesis, and in vitro selection. Selection of mutations previously associated with zidovudine resistance can be an important mechanism through which HIV may escape stavudine. The effect of these mutations on phenotypic stavudine susceptibility is relatively small but apparently large enough to be clinically significant. PMID- 11930318 TI - Feline immunodeficiency virus infection is characterized by B7+CTLA4+ T cell apoptosis. AB - The B7.1 and B7.2 costimulatory molecules on antigen-presenting cells provide second signals for regulating T cell immune responses via CD28 and cytotoxic T lymphocyte antigen 4 (CTLA4) on T cells. CD28 signals cell proliferation, whereas CTLA4 signals for anergy or apoptosis, terminating the immune response. Because T cell apoptosis and immunodeficiency is a characteristic of feline immunodeficiency virus (FIV)-infected cats, it is possible that negative T cell signaling via B7 and CTLA4 may be favored in these cats. Flow cytometry revealed high percentages of CD8+ and CD4+ cells expressing B7.1, B7.2, and CTLA4 in lymph nodes of FIV-positive cats and a large fraction of CTLA4+ T cells coexpressing B7.1 and B7.2. Three-color analysis with anti-B7.1, anti-B7.2, or anti-CTLA4 and TUNEL (terminal deoxynucleotidyl transferase nick-end-labeling) analysis revealed that apoptosis was a characteristic of B7.1+ B7.2+ CTLA4+ T cells. These data support the hypothesis that lymph node apoptosis and immune deterioration in FIV infected cats results from chronic B7.1- and/or B7.2-CTLA4-mediated T-T interactions. PMID- 11930319 TI - Evidence for differential expression of candida albicans virulence genes during oral infection in intact and human immunodeficiency virus type 1-transgenic mice. AB - To comprehensively assess the in vivo expression of Candida albicans hydrolytic enzyme genes during oropharyngeal candidiasis (OPC), a controlled sequential analysis of the temporal expression of individual members of the SAP (secretory aspartyl proteinase) gene family and PLB1 (phospholipase B) in a murine model of OPC was conducted. Acute infections in intact C3H and DBA/2 mice were terminated by clearance of C. albicans within 7 days after oral inoculation, but transgenic (Tg) mice expressing human immunodeficiency virus type 1 were persistently colonized until a final outgrowth before death. In contrast to the sustained expression of other SAP genes and PLB1, SAP7 and SAP8 were conspicuously distinguished by their transient expression in both intact and Tg mice. SAP5 and SAP9 were most strongly expressed throughout the course of infection in the Tg mice. These findings indicate that expression of individual members of the C. albicans SAP gene family is differentially regulated during experimental OPC. PMID- 11930320 TI - Mucosal candidiasis in transgenic mice expressing human immunodeficiency virus type 1. AB - The availability of CD4C/HIV(MutA) transgenic (Tg) mice expressing human immunodeficiency virus type 1 in immune cells and developing an AIDS-like disease has provided the opportunity to devise a model of mucosal candidiasis that closely mimics the clinical and pathologic features of candidal infection in human AIDS. After intraoral infection with Candida albicans, oral burdens were strikingly elevated in the Tg mice, compared with non-Tg littermates (P<.05), during primary infection, a 6-10-week carrier state, and a marked terminal outgrowth preceding death. The chronic carrier state was absent in the non-Tg mice because of clearing of C. albicans. Candida hyphae penetrated the epithelium of the oral cavity, esophagus, and cardial-atrium fold of the stomach, accompanied by a mononuclear cell infiltrate. Immunohistochemical analysis suggested that decreased frequencies of major histocompatibility complex class II expressing cells, combined with reduced CD4+ cells, may underlie the susceptibility to mucosal candidiasis in these Tg mice. PMID- 11930321 TI - Glutathione protects mice from lethal sepsis by limiting inflammation and potentiating host defense. AB - Neutrophils have a dual role in sepsis-defending against infection and mediating organ failure. Because glutathione (GSH) is lower in sepsis, the hypothesis that GSH depletion might impair the migratory response of neutrophils to infection was tested. In a mouse model of polymicrobial sepsis induced by cecal ligation and puncture, GSH depletion inhibited peritoneal neutrophil infiltration, increased bacterial colonies, augmented pulmonary neutrophil infiltrate, and worsened survival. The reduced peritoneal influx of neutrophils was explained by a reduced in vivo neutrophil migration in response to locally administered chemokines and by reduced chemotactic activity and chemokine levels in peritoneal lavage fluid. Conversely, the GSH precursor N-acetyl-l-cysteine augmented neutrophil infiltration in the peritoneum but not in the lung, decreased bacterial colonies, and improved survival. Thus, migration of neutrophils to a site of infection and to a distant site is differently regulated, and optimal GSH levels are important for an efficient response to sepsis. PMID- 11930322 TI - Genogrouping and incidence of virulence factors of Enterococcus faecalis in liver transplant patients differ from blood culture and fecal isolates. AB - Enterococcus faecalis is a leading cause of infections in liver transplant patients. This study reviewed the incidence of virulence factors such as hemolysin, gelatinase, aggregation substances (asa1 and asa373), or the enterococcal surface protein (Esp) in isolates from liver transplant patients. In total, 133 isolates from liver transplant patients were compared with 47 isolates from feces of healthy volunteers and 66 isolates from blood cultures. Amplified fragment length polymorphism (AFLP) analysis indicates that the isolates from different clinical subgroups can be divided into genogroups with an AFLP similarity of >80% and different virulence factors. Hemolysin and asa1 might be associated with infection, as they are more frequent in isolates from blood cultures and transplant patients. Esp might be associated with colonization and spread, because it is more frequent in isolates from feces of healthy volunteers and transplant patients. An epidemic esp gene-positive strain among liver transplant patients supports this hypothesis. PMID- 11930323 TI - Presence of Rickettsia helvetica in granulomatous tissue from patients with sarcoidosis. AB - In samples obtained during the autopsies of 2 patients with sarcoidosis, genetic material from Rickettsia helvetica was detected by polymerase chain reaction, and histologic and immunohistochemical examination (using 3 different antibodies) of the polymerase chain reaction-positive tissues showed different degrees of granuloma formation and presence of rickettsia-like organisms predominantly located in the endothelium and macrophages. Electron microscopic examination clearly identified and demonstrated rickettsia-like organisms within the granuloma, with findings suggestive of ongoing infection. Immunogold labeling with Proteus OX-19 antiserum showed that the gold markers were localized to the rickettsia-like organisms. Paraffin-embedded biopsy specimens from 30 patients with confirmed sarcoidosis were also reexamined, and 26 specimens were judged to be positive for rickettsia-like organisms by histologic and immunohistochemical examination. In a specimen from 1 patient, rickettsia-like organisms also were demonstrated and identified by transmission electron microscopy. These results support the hypothesis that rickettsiae may contribute to a granulomatous process, as is seen in sarcoidosis. PMID- 11930324 TI - Type II alveolar cells play roles in macrophage-mediated host innate resistance to pulmonary mycobacterial infections by producing proinflammatory cytokines. AB - Roles of type II pneumocytes in macrophage (Mphi)-mediated host resistance to pulmonary Mycobacterium tuberculosis (MTB) and M. avium complex (MAC) infections were studied. Electron microscopy of the lung sections of mice given intratracheal infection indicated that the organisms invaded both Mphis and type II pneumocytes. When Mono-Mac-6 Mphis(MM6-Mphis) and A-549 type II pneumocytes (A 549 cells) were cocultivated, bacterial growth in MM6-Mphis was reduced by A-549 cell-derived soluble factors, indicating the roles of type II pneumocytes in Mphi mediated host resistance to mycobacteria. MTB- or MAC-infected A-549 cells showed increased mitochondrial RNA expression of cytokines and surfactant proteins (SPs), in the order tumor necrosis factor-alpha (TNF-alpha) > or = granulocyte Mphi colony-stimulating factor (GM-CSF) > Mphi chemoattractant protein > or = interleukin-8 > SP-D. Anti-TNF-alpha and anti-GM-CSF antibodies attenuated A-549 cell-dependent inhibition of intramacrophage mycobacteria, indicating their crucial roles in A-549 cell-mediated potentiation of Mphi antimycobacterial activity. PMID- 11930325 TI - Onchocerca volvulus-exposed persons fail to produce interferon-gamma in response to O. volvulus antigen but mount proliferative responses with interleukin-5 and IL-13 production that decrease with increasing microfilarial density. AB - The cellular immune response to Onchocerca volvulus antigen (OvAg) was studied in 551 persons exposed to O. volvulus transmission in a hyperendemic area of Ghana, West Africa. A whole-blood assay showed that, in response to a soluble O. volvulus extract, cell proliferation, as well as interleukin (IL)-5 and IL-13 concentrations in the supernatants, were high in cultures of blood from microfilaria (mf)-negative persons and significantly decreased with increasing mf counts of the donors. Only background concentrations of interferon (IFN)-gamma were found, and these did not correlate with mf counts. In response to a mitogen, cells from all persons strongly reacted with proliferation and secretion of all 3 cytokines studied. These findings show that the response of human peripheral blood cells to OvAg does not include significant IFN-gamma production; that mf negativity is associated with IL-5 and IL-13 production but not, as previously suggested, with IFN-gamma production; and that IL-5 and IL-13 production decreases with increasing mf density. PMID- 11930326 TI - Protection of humans against malaria by immunization with radiation-attenuated Plasmodium falciparum sporozoites. AB - During 1989-1999, 11 volunteers were immunized by the bites of 1001-2927 irradiated mosquitoes harboring infectious sporozoites of Plasmodium falciparum (Pf) strain NF54 or clone 3D7/NF54. Ten volunteers were first challenged by the bites of Pf-infected mosquitoes 2-9 weeks after the last immunization, and all were protected. A volunteer challenged 10 weeks after the last immunization was not protected. Five previously protected volunteers were rechallenged 23-42 weeks after a secondary immunization, and 4 were protected. Two volunteers were protected when rechallenged with a heterologous Pf strain (7G8). In total, there was protection in 24 of 26 challenges. These results expand published findings demonstrating that immunization by exposure to thousands of mosquitoes carrying radiation-attenuated Pf sporozoites is safe and well tolerated and elicits strain transcendent protective immunity that persists for at least 42 weeks. PMID- 11930327 TI - Neutralization of pseudotyped vesicular stomatitis virus expressing hepatitis C virus envelope glycoprotein 1 or 2 by serum from patients. AB - Infection with hepatitis C virus (HCV) generally progresses to chronic disease, although a minority of patients appear to clear viremia spontaneously. In this investigation, serum samples were analyzed for virological parameters, serum alanine aminotransferase (ALT) levels, and neutralizing antibody response against pseudotyped vesicular stomatitis virus (VSV) generated using chimeric envelope glycoprotein 1 (E1) or 2 (E2) of HCV. Testing of sequential serum samples that were collected beginning at the onset of acute-phase disease demonstrated intermittent viremia, elevated ALT levels, and detectable neutralization activity against VSV in 9 of 10 patients. Serum neutralization activity did not exhibit a correlation with the genotype of the infecting HCV or with virus load. On the other hand, patients with chronic HCV infection consistently had detectable amounts of virus present but no significant variation in ALT levels, and serum samples from a majority (>90%) of patients failed to show detectable neutralization activity. PMID- 11930328 TI - Hepatitis A virus infection in tamarins: experimental transmission via contaminated factor VIII concentrates. AB - An experimental hepatitis A virus (HAV) transmission study was performed in 3 tamarins, using a factor VIII concentrate linked to a recent outbreak of HAV infections in German hemophiliacs. The typical indicators for HAV infection were investigated in feces and serum samples. One tamarin showed a classical HAV infection with seroconversion. HAV antigen and HAV RNA were detected in feces of a second animal, but no seroconversion was observed until 19 weeks after inoculation. The HAV sequences from the reverse-transcription- and polymerase chain reaction-positive samples of the 2 animals were identical to the deduced HAV sequences of the chain of infection (from plasma pool to final product to patients). The study results provide conclusive evidence of the presence of infectious HAV in coagulation factor concentrate. Because a number of HAV transmission episodes have been described for solely solvent/detergent-treated factor VIII preparations, continued use of these agents seems questionable. PMID- 11930329 TI - Monocyte chemoattractant protein-2 (CC chemokine ligand 8) inhibits replication of human immunodeficiency virus type 1 via CC chemokine receptor 5. AB - CC chemokine receptor 5 (CCR5) is a coreceptor for cellular entry of monocyte tropic (R5) strains of human immunodeficiency virus (HIV) type 1, which has been implicated as the predominant phenotype of HIV in early infection. The CCR5 agonists macrophage inflammatory protein (MIP)-1alpha, MIP-1beta, and RANTES (regulated on activation, normally T cell-expressed and -secreted) have been shown to block replication of R5 virus in vitro and have gained attention as potential antiviral factors. However, a few reports have suggested that other chemokines may also block R5 HIV-1, including monocyte chemoattractant protein (MCP)-2 (CC chemokine ligand 8). We demonstrate that MCP-2 specifically inhibits replication of R5 HIV-1 and that this activity is additive to that of RANTES. Furthermore, MCP-2 induces a robust, pertussis toxin-sensitive calcium flux in primary lymphocytes, and cross-desensitization studies indicate that MCP-2 acts via CCR5. These data confirm that MCP-2 is a ligand for CCR5 on CD4(+) lymphocytes and can specifically block R5 HIV-1. PMID- 11930330 TI - A pilot study of the discontinuation of antifungal therapy for disseminated cryptococcal disease in patients with acquired immunodeficiency syndrome, following immunologic response to antiretroviral therapy. AB - To determine whether microbiologic cure of acquired immunodeficiency syndrome (AIDS)-related disseminated cryptococcosis is possible in patients receiving highly active antiretroviral therapy (HAART), antifungal therapy was discontinued in 6 patients with a history of disseminated cryptococcosis who had received > or =12 months of antifungal therapy. All were asymptomatic and had absolute CD4+ T cell counts of >150 cells/microL (range, 178-525 cells/microL). Blood, cerebrospinal fluid (CSF), and urine samples were obtained for fungal culture. Serum and CSF cryptococcal antigen titers were also obtained. All 6 patients had CSF and blood cultures negative for Cryptococcus neoformans and were receiving HAART. All patients' subsequent cultures remained sterile, and all patients were clinically asymptomatic 24 months after ending antifungal therapy. Disseminated cryptococcal disease can be cured by prolonged antifungal therapy in some patients with AIDS who experience sustained CD4 lymphocyte increases while receiving HAART. PMID- 11930331 TI - Protective effect of interleukin-4 -589T polymorphism on human immunodeficiency virus type 1 disease progression: relationship with virus load. AB - The interleukin (IL)-4 -589T allele bears a single nucleotide polymorphism at position -589 upstream from the open-reading frame of the IL-4 gene. To determine the influence of this allele on human immunodeficiency virus (HIV) type 1 disease, disease progression and serum virus load were assessed by IL-4 genotype in 427 white patients with known seroconversion dates who were followed in the French SEROCO cohort between 1988 and 1996. Serum virus load was 0.20 log lower during the 6-24-month plateau phase after seroconversion in patients with IL-4 589T than in those without this allele (P=.02). Kaplan-Meier analysis survival curves showed a slower progression to clinical AIDS in carriers of IL-4 -589T (P=.04). Adjustment for early serum virus load greatly diminished the strength of this association. These results suggest that IL-4 -589T protects against HIV-1 disease progression by reducing virus load. PMID- 11930332 TI - Vitamin A supplementation and human immunodeficiency virus type 1 shedding in women: results of a randomized clinical trial. AB - Observational studies have associated vitamin A deficiency with vaginal shedding of human immunodeficiency virus (HIV) type 1-infected cells and mother-to-child HIV-1 transmission. To assess the effect of vitamin A supplementation on vaginal shedding of HIV-1, a randomized, double-blind, placebo-controlled trial of 6 weeks of daily oral vitamin A (10,000 IU of retinyl palmitate) was conducted among 400 HIV-1-infected women in Mombasa, Kenya. At follow-up, there was no statistically significant difference in the prevalence of HIV-1 DNA (18% vs. 21%, P=.4) or the quantity of HIV-1 RNA (3.12 vs. 3.00 log(10) copies/swab, P=1.0) in vaginal secretions of women receiving vitamin A, compared with women receiving placebo. No significant effect of supplementation on plasma HIV-1 load or CD4 or CD8 cell counts was observed, and no effect was seen among women who were vitamin A deficient at baseline. Vitamin A supplementation is unlikely to decrease the infectivity of women infected with HIV-1. PMID- 11930333 TI - Immune protection against staphylococcal enterotoxin-induced toxic shock by vaccination with a Venezuelan equine encephalitis virus replicon. AB - A candidate vaccine against staphylococcal enterotoxin B (SEB) was developed using a Venezuelan equine encephalitis (VEE) virus vector. This vaccine is composed of a self-replicating RNA, termed "replicon," containing the VEE nonstructural genes and cis-acting elements and a gene encoding mutagenized SEB (mSEB). Cotransfection of baby hamster kidney cells with the mSEB replicon and 2 helper RNA molecules resulted in the release of propagation-deficient mSEB-VEE replicon particles (mSEB-VRPs). Mice inoculated subcutaneously with mSEB-VRPs were protected (15 of 20 mice) from a challenge with 5 median lethal dose units of wild-type (wt) SEB. T cells from mice vaccinated with mSEB-VRP responded normally both in vitro to wt SEB and in recall response to the inactivated mSEB polypeptide. The profile of cytokines measured after challenge with wt SEB suggested that the mode of protection was predominantly Th1 dependent. Our results suggest that the VEE replicon is a practical and convenient model system for evaluating efficacy of vaccines for the control of bacterial diseases. PMID- 11930334 TI - Worldwide incidence of multidrug-resistant tuberculosis. AB - Planning for tuberculosis (TB) control requires an assessment of the number and distribution of drug-resistant cases. This study used results of resistance surveys from 64 countries, together with data predictive of resistance rates from 72 others, to estimate the number of new multidrug-resistant (MDR) TB cases that occurred in 2000. By these methods, an estimated 273,000 (95% confidence limits, 185,000 and 414,000) new cases of MDR TB occurred worldwide in 2000, 3.2% of all new TB cases. The analysis provides the first comprehensive set of estimates of the MDR TB burden by country and globally. PMID- 11930335 TI - Polymerase chain reaction detection of aspergillus DNA in experimental models of invasive aspergillosis. AB - To determine the sensitivity of polymerase chain reaction (PCR) assays for the diagnosis of invasive aspergillosis, results of quantitative culture, PCR-ELISA, and a quantitative LightCycler assay (Roche Diagnostics) of blood and organ specimens of experimentally infected mice and rabbits were compared. By PCR ELISA, 297 of 379 murine lung specimens were positive, but only 235 of 379 were culture positive. Whereas 64 culture-negative lungs were positive by PCR, Aspergillus was grown from only 2 PCR-negative samples. The PCR assay was 19.4 times more sensitive than culture. None of the 68 blood cultures from mice and rabbits were positive for Aspergillus fumigatus, whereas PCR detected Aspergillus DNA in 17 of 68 blood samples. Quantitative PCR analysis of blood samples showed a fungus load of 10(1)-10(2) cfu/mL of blood. The data confirm the superior sensitivity of PCR for the diagnosis of experimental Aspergillus infections. PMID- 11930336 TI - Identification of a conserved Plasmodium falciparum var gene implicated in malaria in pregnancy. AB - The Plasmodium falciparum erythrocyte membrane protein 1 (PfEMP1) family is a highly polymorphic class of variant surface antigens encoded by var genes that play an important role in malaria pathogenesis. This report describes the unexpected finding that 1 of the var genes encoding a PfEMP1 variant that binds to the host receptor chondroitin sulfate A (CSA) and is implicated in malaria in pregnancy is well conserved among P. falciparum isolates worldwide. The N terminal domains of this PfEMP1 variant are especially highly conserved, whereas the functional CSA binding domain is more variable. Analysis of var gene expression in placental parasites from primigravid women in Malawi did not support a role for this conserved gene in placental infection but identified a second commonly occurring var gene. These results indicate the need for reevaluation of previous assumptions of a minimal overlap between var gene repertoires from different parasite isolates. PMID- 11930337 TI - Tobacco cessation intervention in a nurse practitioner managed clinic. AB - Tobacco cessation counseling by health care professionals has been associated with increased tobacco cessation rates. In this study we compare the effectiveness of two smoking cessation approaches administered by nurse practitioners--a stepped care approach (n = 34) and a routine care approach (n = 41), using a pre- and post-test control group study design. Additionally, this study was guided by the "stages of change" construct of the transtheoretical model. Study results indicated that nurse practitioners delivered consistent office-based advice about tobacco use. In addition, nurse practitioner smoking cessation interventions contributed to positive shifts in stage of change for smoking cessation. PMID- 11930338 TI - Body piercing: what nurse practitioners need to know. AB - Body piercing is a growing trend among today's youth. Chances are, nurse practitioners will be in contact with someone who has a body piercing and/or complication relating to piercing. This article includes the history of body piercing, the environment of piercing parlors, the methods commonly used, site specific explanations of select piercings, complications, treatment and practice implications. Nurse practitioners need to establish collaborative working relationships with their clients as well as those who do the piercings. PMID- 11930339 TI - Differential diagnosis of infective endocarditis. AB - Infective endocarditis (IE) is a deadly disease if left untreated. Early detection and treatment of IE reduces the high mortality rate. Mitral valve prolapse is now the leading risk factor for development of IE. The Duke (1994) criteria includes echocardiography as a diagnostic tool. Outclient i.v. therapy is increasing due to once-daily dosing regimens, which reduces the cost of treatment. Identifying at risk clients and educating them is vital to the prevention of IE. This article familiarizes the primary care provider with the pathophysiology, clinical presentation, diagnostic testing, complications, medical treatment, prophylactic management, and client education of IE. PMID- 11930340 TI - [CAD/CAM-copings for partial coverage]. AB - Aim of the study was to evaluate the Computer Integrated Ceramic Reconstruction (CICERO)-system and the Ceramic Reconstruction (CEREC)-system for the production of all ceramic copings for partial coverage. Posterior teeth were prepared and the stone dies were made. Accuracy analyses were performed on ceramic restorations made by means of the CICERO and by means of the CEREC technique. The marginal gaps were compared to that of a control cast metal restoration. The results demonstrate that the marginal gaps of the CICERO and CEREC copings varied respectively for the premolar 58-80 microns (mean 69 microns) and 71-91 microns (mean 81 microns). For the upper molar 63-92 microns (mean 78 microns) and 68-110 microns (mean 89 microns) and for the lower molar 54-98 microns (mean 76 microns) and 73-99 microns (mean 86 microns). Control cast metal partial coverage restorations showed marginal gaps of 33, 49 and 41 microns. It is concluded that computers can produce copings for partial coverage preparations on stone dies with a mean marginal gap for CICERO copings of 74 microns and for CEREC copings of 85 microns. These values were obtained before optimizing the marginal fit by means of porcelain veneering. PMID- 11930341 TI - [What does the non-dentistry-trained person know about avulsion?]. AB - The prognosis of an avulsed permanent incisor depends on a swift and correct emergency treatment. The chance of a favourable prognosis increases if the tooth is replanted as quickly as possible, preferably by the persons first present at the scene of the accident. Usually this person is a layman, therefore the aim of this study is to investigate the knowledge in this respect. To 48 schools and 15 swimming pools in Amsterdam, Arnhem and Nijmegen (the Netherlands), questionnaires were sent. Thirty-six schools and 15 swimming pools responded. Twenty-eight percent of the respondents had previous experience with an avulsion. Ten percent of the respondents would replant the incisor and 90% would store the incisor. Sixty-three percent would treat the tooth correctly. Respondents with previous experience had no more knowledge of the most effective treatment than persons without previous experience. Swimming instructors and physical health instructors knew significantly more about avulsion than teachers in primary schools. Since only 10% of the respondents would replant the incisor and 37% would handle this accident incorrectly, more information seems to be necessary. PMID- 11930342 TI - [Ergonomics in dental practice. Prevention of physical and mental overload]. AB - In this article two important issues in dental ergonomics are discussed: physical and mental stress. It is important for a dentist to stay aware of the balance between on one hand short term profit against various forms of stress, and on the other hand long term negative effects. Possible problems are described without going in more deeply into theoretical backgrounds. Practical suggestions for action are given for the dentist who wants to work with pleasure and in good health over a long period of time. PMID- 11930343 TI - [Syndromes 8. The diagnosis of congenital anomalies and syndromes]. AB - In 2 to 3% of newborns a congenital anomaly is detected. About 20% of these have even multiple anomalies. After a short review on the classification of congenital anomalies a description of the diagnostic workup is given with emphasis on items which are different from the usual clinical practice: pregnancy history, family history, dysmorphologic examination, use of electronic databases and genetic/molecular investigations. The aim of this workup is to reach an etiologic diagnosis which is a prerequisite for both establishing prognosis and adequate therapy and genetic counseling. PMID- 11930344 TI - [NVT-Fall Congress: 'From mutilation to rehabilitation']. PMID- 11930345 TI - [A normal case of impacted molars?]. PMID- 11930346 TI - [Alzheimer's disease 1]. PMID- 11930347 TI - [Alzheimer's disease 2]. PMID- 11930348 TI - [Bone transplantation in cleft palate patients. A retrospective study]. AB - Bone-grafts were evaluated in 45 patients with uni- or bilateral residual alveolar clefts. In this population there were 31 unilateral clefts and 14 bilateral clefts. This makes a total of 59 bone grafts. The mean follow-up period was 5.7 years. The patients were divided into two groups. Group A (grafting prior to the eruption of the homolateral canine) and group B (grafting after the eruption of the homolateral canine). Group A consisted of 38 grafts in 30 patients and group B consisted of 21 grafts in 15 patients. The grafts were harvested from the iliac crest. The alveolar bone-height was evaluated on occlusal radiographs. Results in patients treated prior to the eruption of the homolateral canine were significantly better then in patients treated after eruption of the canine. No difference was found between unilateral or bilateral clefts. Of the 38 canines which had not erupted at the time of the grafting procedure 26 (68.4%) erupted spontaneously. PMID- 11930349 TI - [Preparation with air abrasion]. AB - Air abrasive technology has re-emerged in dentistry. The history, characteristics and clinical application of this approach are reviewed, including advantages and limitations for the removal of enamel, dentin and decay. Air-abrasive technology may be especially suited for use in bonded restorations as well as for repair of all types of restorations. The factors affecting adequate bond strength are discussed. PMID- 11930350 TI - [Syndromes 9. Hemifacial microsomia]. AB - Hemifacial microsomia is also known under quite a number of synonyms. Pathognomonic for these usually sporadic cases are: marked three dimensional asymmetry of the mandible, the ear, the maxilla, the zygoma and the orbit. The more distal the structure, the more marked the asymmetry. Many other optional symptoms are described. The differential diagnosis is almost never a problem in this aspecific complex of symptoms. Surgical therapy asks for expert treatment. The treatment contains usually several operations of which one to three are really major. Even then, very good results are mainly obtained in the less pronounced cases only. PMID- 11930351 TI - [Risks of ingesting cleansing tablets for dental prostheses]. AB - Aim of the study was to determine the frequency and severity of intoxication caused by cleansing tablets for dental prostheses. The investigation was performed by the National Poisons Controle Centre in Bilthoven, the Netherlands. In 1996, alle 78 physicians who requested information from the Centre about intoxications with cleansing tablets for dental prostheses were sent a questionnaire enquiring about the exposure situations and the severity of the effects of the intoxications. Forty-six questionnaires (59%) were returned. Older people (24: 52%) and children (14; 30%) constituted the largest groups of intoxicated. As a rule, accidental ingestion of the tablets was involved. In most cases, one tablet had been swallowed. Local effects, mostly of a mild nature, were reported in 22 patients. Systemic effects were reported in two patients but in these cases the connection with the ingestion was doubtful. PMID- 11930352 TI - [Letters from Nederlands Tijdschrift voor Geneeskunde]. PMID- 11930353 TI - [Why do the upper incisors not erupt?]. PMID- 11930354 TI - [Oral carcinoma in 34 dogs]. AB - In a retrospective study, the epidemiologic data, treatment results and survival times of dogs with an oral squamous cell carcinoma in the period 1983-1996 have been reviewed, using the data base of the Department of Clinical Sciences of Companion Animals of the Faculty of Veterinary Medicine of the University of Utrecht, The Netherlands. The etiology of oral squamous cell carcinoma in dogs is unknown. For various reasons, such as extent of the disease, active treatment was undertaken in only 22 out of 34 dogs. Two dogs were lost during follow-up. Sixteen of the remaining 20 dogs who underwent surgical treatment for their oral squamous cell carcinoma were alive, one year postoperatively, apparently without tumour. Postoperative radiotherapy may even improve the survival rates. PMID- 11930355 TI - [Syndromes 10. Beckwith-Wiedemann syndrome]. AB - Beckwith-Wiedemann syndrome is characterised by variable clinical combinations of exomphalos, macroglossia, gigantism, hypoglycemia and an increased incidence of tumors. This article briefly describes its clinical symptomatology. Recently, hemangiomatous lesions in tongues have been determined. PMID- 11930356 TI - [New direct restorative materials]. AB - People worldwide have become increasingly aware of the potential adverse effects on the environment, of pollution control and of toxic effects of food, drugs and biomaterials. Amalgam and its potential toxic side effects (still scientifically unproven) continue to be discussed with increasing controversy by the media in some countries. Consequently, new direct restorative materials are now being explored by dentists, materials scientists and patients who are searching for the so-called 'amalgam substitute' or 'amalgam alternative'. From a critical point of view some of the new direct restorative materials are good with respect in aesthetics, but all material characteristics must be considered, such as mechanical properties, biological effects, and longterm clinical behaviour. PMID- 11930357 TI - [A recurrent swelling of the gingiva]. PMID- 11930358 TI - [Parkinson disease]. PMID- 11930359 TI - [Etiology of missing teeth]. AB - This article gives a brief overview of the causes of tooth loss. Caries is still the most prominent reason for tooth loss. In patients over 40 years, periodontal reasons become more prevalent. However, also in this age category caries is the main reason. In young individuals, missing teeth are most frequently associated with agenesia or trauma. Iatrogenic damage is considered to be a casuistic cause of tooth loss. Nevertheless, some specific areas, such as in the combined treatment 'endodontic treatment--post and core--crown', can be distinguished in which tooth loss is more frequently seen. PMID- 11930360 TI - [Prevention of dental trauma]. AB - Dental trauma is for people under 25 years of age a larger threat than caries and periodontal diseases. Protrusion of the upper front teeth in youth should therefore be corrected by orthodontic treatment. By doing contact-sports the use of the mouth guard is necessary. Since several different types are available, one has to make a correct choice specified by the kind of sport. The statement that it is more important to use an apparatus than just to own it sounds logically, but is often not followed in daily live. PMID- 11930361 TI - [The single missing tooth. Treatment possibilities from an orthodontic view point]. AB - In patients with single missing teeth, a standard treatment plan does not exist. The treatment plan depends on the age and the facial profile, the malocclusion, the interdigitation, the available space, the amount of bone, the shape, color, and the level of the gingiva of the neighbouring teeth. Closing or opening the diastema should be considered. A diagnostic set-up of the final occlusion can provide more insight into the final result that can be expected from the treatment modalities. Orthodontic closure of the diastema is preferable, but not always feasible or desirable and compensatory extractions may be necessary. In children, auto-transplantation is an option. In adults with a sufficient amount of bone the placement of an implant may be the treatment of choice. Frequently, preimplantological bone augmentation is necessary because otherwise prosthodontic replacement is the only solution. PMID- 11930362 TI - [Single tooth replacement with a removable partial denture]. AB - The indication of removable partial dentures in cases of single tooth replacement is limited. This is due to a number of disadvantages closely connected to the use of removable partial denture. An often applied use is that of a mucosa-borne transitional acrylic denture to improve patients aesthetics until the definitive denture can be made. Acrylic dentures may be indicated as a definitive denture when limited financial resources are available. A metal frame removable partial denture can be applied to lengthen a shortened dental arch or in specific cases in which fixed partial dentures are not the preferable treatment option. The success of the treatment with a removable partial denture is highly dependent on the follow-up service and the oral hygiene awareness. PMID- 11930363 TI - [Three-unit bridge prostheses. With or without crowns?]. AB - Replacement of a single tooth by a fixed partial prosthesis is possible in several ways: a conventional bridge, a resin-bonded bridge or a UDA-bridge (Universal Dental Anchorage). Preservation of natural tooth substance, an optimal environment for the periodontal tissues and good esthetics are the main criteria for making a choice. Unrestored or minimally restored front teeth are an indication for a resin-bonded bridge. When an indication for a crown on the adjacent teeth is already present, a conventional bridge is the treatment of choice. Although the future promises metal free bridge solutions, the long-term prognosis of posterior bridges is best guaranteed by using metal-ceramic bridges. The UDA system requires improvement for general use. PMID- 11930364 TI - [Single tooth replacement with an implant. Possibilities and limitations]. AB - Implant treatment in case of a single missing tooth can potentially lead to a well functioning and aesthetically pleasing single crown on an implant. However, in order to achieve an optimal treatment result, a long road has to be travelled since usually resorption defects have to be repaired and soft tissue corrections are frequently necessary. This paper provides information regarding commonly observed situations and their consequences for the prognosis of implant treatment. It is concluded that a single tooth implant can be an elegant solution, though not an easy one. It requires a high degree of experience and surgical and prosthetic capabilities of the dentist. The natural situation can only be mimicked if all stages of treatment are adequately planned and conducted and if no major complications occur in the process. To prevent disappointments, both dentist and patient should be well aware of all conditions required for an optimal result. PMID- 11930366 TI - [Local bone augmentations for the use of implants]. AB - Placement of endosseous implants is often restricted because of lack of supporting bone, limiting placement of implants of adequate length in a prosthodontically optimal position. Several surgical procedures have been described to create sufficient volume of bone for placement of implants. In this paper, various techniques are described for harvesting intraoral bone grafts for augmentation of a localized defect of the alveolar ridge. PMID- 11930365 TI - [Preoperative planning for the placement of a single implant]. AB - Radiography is of primary importance for the preoperative planning of oral implant placement. Depending on the indication, various techniques are available, each with its own merits and drawbacks. In general, it can be stated that the intra-oral radiography using the paralleling technique is the method of choice for a solitary tooth replacement. Panoramic radiography is indicated for implant placement in the symphyseal area of the lower jaw. Cross-sectional imaging is performed when information is needed in the bucco-lingual dimension. Conventional tomography is recommended for preoperative planning of limited edentulous areas. CT-scan imaging is required for rehabilitation of extended edentulous areas or in complex maxillofacial surgery combined with implant placement. Choosing the optimal technique can be performed by weighing the radiation dosage and the diagnostic information required. PMID- 11930367 TI - [Guided bone regeneration for single tooth replacements by oral implants]. AB - Single tooth replacement is a good indication for an oral implant. However, bone regeneration, is often required to improve the bone quantity and the contour of the overlying soft tissues. Guided Bone Regeneration (GBR) using membranes is a good technique to restore local bone defects within a six months period. With this technique it is crucial that a membrane properly separates the bony and soft tissues and creates a space around a bony defect for bone ingrowth. This article describes the indications and contra-indications for GBR, the technique and the possible complications. PMID- 11930368 TI - [Immediate repair of labial bone defects]. AB - In this paper some surgical techniques are described to prevent or restore the characteristic and esthetically disturbing resorption defect in the anterior alveolar process as a result of tooth loss. One-stage approaches are described. Submucosal implants are used in combination with conventional fixed prosthodontics. If a single tooth replacement by means of an implant is aimed at, the split-crest technique may be a valuable alternative for an autologous bone graft or guided bone regeneration procedure. PMID- 11930369 TI - [Considerations of single tooth replacement. The choice between different treatment options]. AB - When a single tooth is lost both functional and aesthetic aspects have to be restored. A fixed prosthodontic restoration such as a conventional bridge, a resin-bonded bridge or a solitary implant is the treatment of choice. The final choice between these three treatment modalities strongly depends upon specific characteristics in the patients' dentition and his preferences. There is a vast amount of clinical experience with the conventional fixed bridge and it is available in every dental office. Without a doubt the resin-bonded bridge is the cheapest option. However, the patient does run the risk of loosening of the bridge, usually at an inconvenient moment. It then has to be recemented or replaced. The solitary implant has several advantages over the fixed and resin bonded bridge. For optimal results, it does require a physician with a vast experience in the field of oral implantology. For young individuals closure of the diastema by means of orthodontics is also possible. A partial denture is only an option as a temporary provision or when more teeth were lost or are bound to be lost. PMID- 11930370 TI - [Five year survival of posterior adhesive bridges. Influence of bonding systems and tooth preparation]. AB - The present study reports the final analysis of a randomized controlled clinical trial in which different designs of posterior resin-bonded bridges were evaluated for a period of at least 5 years. The operational hypothesis was that the bonding system and the preparation design used in posterior resin-bonded bridges have an influence on the survival and clinical functioning of these restorations. Survival in this study was defined at two levels: (1) 'primary' survival (survival without any debonding), and (2) 'functional' survival (survival including loss of retention on one occasion and successful rebonding of the original resin-bonded bridge without further debonding). Preparation of grooves in abutment teeth for posterior resin-bonded bridges appeared to be beneficial to their chance of survival. Resin-bonded bridges placed in the maxilla have a better prognosis than those made in the mandible. The bonding systems used in this study (etching/Clearfil F2, sand blasting/Panavia EX and silica coating/Microfill Pontic C) appear to have no influence on the chance of failure with regards to the 'primary' survival. In rebonded posterior resin-bonded bridges, the bonding system silica coating/Microfill Pontic C was more retentive than the other systems tested. PMID- 11930371 TI - [The effect of drugs on the orofacial area]. AB - The oral and maxillofacial region is affected by the negative side effects of a wide variety of endogenic and exogenic factors including drugs. Drugs are commonly used to treat oral and maxillofacial diseases, but more often the oral and maxillofacial region is affected by the side effects of drugs and home remedies on the orofacial tissues taken by the patient himself or prescribed by a dentist or physician. In this paper both factors are described which the dentist has to consider when he prescribes a drug as well as the oral and maxillofacial side effects of a variety of drugs. PMID- 11930372 TI - [Syndromes 12. Turner syndrome]. AB - Turner syndrome is one of the most common chromosomal disorders. The incidence is about 1 on 2,500 till 1 on 10,000 living female young births. Short stature is the most common finding in patients with Turner syndrome. Besides short stature and gonadal dysgenesis, typical craniofacial and dental features are also present. Disturbance of the enchondral ossification results in abnormal craniofacial morphology. Oestrogen medication, to induce their puberty, and recombinant human growth therapy, to improve final height of these patients, are the most common treatment possibilities. It is the intention of this short paper to inform the dentist/orthodontist about the general aspects of the Turner syndrome. This information can be used in their treatment plan. PMID- 11930373 TI - [NVT-Spring Congress: 'Future for eighty year olds with teeth']. PMID- 11930374 TI - [Retention costs 3 molars]. PMID- 11930375 TI - [Multiple sclerosis 2]. PMID- 11930376 TI - Intrapartum fetal monitoring: past, present, and future. AB - The concept of intrapartum "monitoring" of the fetal heart rate by auscultation has been extant for almost 200 years and by electronic means for more than 30 years. This article explores historical aspects of fetal monitoring, the advent of electronic fetal monitoring and its controversies, and present and future research opportunities to enhance the reliability, validity, and efficacy of fetal monitoring. PMID- 11930377 TI - The physiology of fetal heart rate patterns and perinatal asphyxia. AB - The purpose of electronic fetal heart rate (FHR) monitoring is the ongoing assessment of fetal oxygenation. FHR tracings are analyzed for characteristic patterns that signify specific hypoxic or nonhypoxic events. A working knowledge of fetal physiology and the fetal response to hypoxia can aid and refine clinical interpretation of FHR patterns during labor. This article reviews the fetal response to decreased oxygenation, the physiology of subsequent FHR patterns and the clinical presentation of asphyxia in the newborn. PMID- 11930378 TI - Risk management and electronic fetal monitoring: decreasing risk of adverse outcomes and liability exposure. AB - Electronic fetal monitoring (EFM) has the potential to promote fetal health and improve neonatal status at birth; however, EFM as a stand-alone tool is ineffective in avoiding preventable adverse outcomes. It is effective only when used in accordance with published standards and guidelines by professionals skilled in correct interpretation and when appropriate timely intervention is based on that interpretation. Interpretation and intervention are best accomplished as a collaborative perinatal team rather than individual activity. Only in these circumstances can EFM optimally contribute to fetal well-being and subsequent neonatal health. Risk management strategies to decrease potential liability are presented that can be accomplished with careful planning and collaboration among perinatal team members. PMID- 11930379 TI - Intermittent auscultation of the fetal heart rate: a review of general principles. AB - Multiple randomized clinical trials have been unsuccessful in providing evidence of efficacy of electronic fetal monitoring; thus, there is renewed interest in intermittent auscultation during labor for women with low-risk pregnancies. Auscultation must be used with palpation or external or internal electronic monitoring of uterine contractions. Auscultation and palpation require education, experience, and competency validation at regular intervals. Institutional policies and standards of care are mandatory for intermittent auscultation. Concerns exist regarding the personnel costs for auscultation; however, these costs may ultimately be shown to be offset by significant benefits in improved outcomes and patient satisfaction. PMID- 11930380 TI - Meta analysis: effects of interventions on premature infants feeding. AB - Nipple feeding of premature infants presents a challenge to neonatal nurses who are trying to prepare the infants for eventual discharge from the hospital. To determine what empirical evidence there was to support interventions that positively influence feeding performance and feeding efficiency a meta-analysis is presented. The effects of NPO, nasogastric tubes, different nipples, breast versus bottle feeding, gestational age, oral support, oral stimulation, and nonnutritive sucking on volume intake are evaluated. The results suggest that nursing care should: (a) consider the infant's gestational age; (b) maturational ability and development; (c) keep track of and attempt to minimize the length of time an infant is NPO; and (d) provide prefeeding oral stimulation and oral support. PMID- 11930381 TI - Developmentally supportive caregiving and technology in the NICU: isolation or merger of intervention strategies? AB - Providing nursing care in the highly technical environment of the neonatal intensive care unit is changing as concepts of developmentally supportive caregiving merge with conventional nursing interventions and are integrated into routine practice. Medical knowledge with the use of technology is insufficient in guiding the whole of neonatal nursing care. Integration of other patterns and concepts is essential to expert nursing practice with high risk infants and their families. PMID- 11930382 TI - Internet resources for perinatal and neonatal nurses. AB - The goal of this column is to provide JPNN readers with a periodic update of additional Internet resources they may find useful for their practice, for recommending to clients and their families, or for teaching. PMID- 11930383 TI - Nurse practitioners' perceptions of their caring behaviors. AB - PURPOSE: To explore nurse practitioners' (NPs) perceptions of their own caring behaviors, the relationship between sociodemographic variables, environmental factors, and NP's perceptions of their caring behaviors. DATA SOURCES: A mailed survey to a systematic random sample of 200 members of an Illinois NP group. CONCLUSIONS: The top ten caring behaviors in rank order were appreciating the patient as a human being, showing respect for the patient, being sensitive to the patient, talking with the patient, treating patient information confidentially, treating the patient as an individual, encouraging the patient to call with problems, being honest with the patient, and listening attentively to the patient. IMPLICATIONS FOR PRACTICE: The quality of instruction in the biomedical aspect of nursing education is relatively easily assessed. Caring is nurses' hidden work that may go unrecognized except when the caring behaviors are missed by the patients or their families. PMID- 11930384 TI - The distribution of advanced practice nurses in Arkansas: gaps in the care of rural underserved populations. AB - PURPOSE: To examine the relationships between advanced practice nurses' (APN) practice sites and economic factors, racial and ethnic characteristics of the populations they serve, and the likelihood of APNs practicing in underserved areas. DATA SOURCES: Retrospective analysis of U.S. census data and records supplied by the Arkansas State Board of Nursing. CONCLUSIONS: Nurse practitioners (NPs) were the only APN group with equal distribution in rural or urban areas. Overall, APNs and NPs were more likely to practice in counties with higher rates of poverty and lower education levels. IMPLICATIONS FOR PRACTICE: More effort needs to be made in recruiting and retaining APNs in rural and underserved areas. PMID- 11930385 TI - Hypercholesterolemia: a look at low-cost treatment and treatment adherence. AB - PURPOSE: To determine whether a positive cholesterol-lowering effect could be achieved with a psyllium dose of 6 grams per day instead of the usual 10 grams per day as advocated by other researchers. DATA SOURCES: Randomized trial of 46 males and females with hypercholesterolemia; multivariate analysis of variance with repeated measures on 1 factor done on 28 subjects (18 in treatment group, 10 in control group) remaining after 16 weeks of treatment. CONCLUSIONS: Lipoprotein analysis at 2, 8, and 16 weeks indicated that a daily dose of 6 grams of psyllium hydrophilic mucilloid did not significantly affect serum total cholesterol nor low-density lipoproteins in either men or women with hypercholesterolemia. The effects of psyllium on hypercholesterolemia appear to be dose dependent. PRACTICE IMPLICATIONS: Although it is a low cost option, the addition of psyllium to the diet has unpleasant side-effects, including abdominal distention, flatulence, and discomfort. Because these side effects are troublesome, the lowest effective dose of psyllium may be an important factor in improving treatment adherence. PMID- 11930386 TI - Child abuse: clinical findings and management. AB - PURPOSE: To provide an overview of the role of the nurse practitioner (NP) in identifying, reporting, and managing child abuse in primary care. DATA SOURCES: Selected research, national guidelines, and the author's experience. CONCLUSIONS: Child abuse is a complex phenomenon characterized by maladaptive behaviors between children and their parents. IMPLICATIONS FOR PRACTICE: The role of the NP includes identification of families at risk, recognition of clinical findings of abuse, diagnosis of abuse, education for families identified at risk, and management of children diagnosed with abuse. The ultimate goal is the safe return of the child to a loving family. PMID- 11930387 TI - Factors contributing to the quality of end-of-life care. AB - PURPOSE: To examine the factors influencing healthcare for the elderly and the terminally ill in modern health care. DATA SOURCES: Selected articles, Internet sources, and legal sources. CONCLUSION: Despite increasing use of advance directives, patients are not given adequate information at an appropriate time to make informed decisions. Even when advance directives are in place, they may not be honored. IMPLICATIONS FOR PRACTICE: Among the most critical areas to consider in improving end-of-life care are encouraging patients to make known their wishes, respecting patients' concerns about quality of life, and promoting continuity and skill in end-of-life care. PMID- 11930388 TI - Post-traumatic stress disorder in primary care practice. AB - PURPOSE: To provide clinicians in primary care settings with guidelines on evaluation, diagnosis, and management of post-traumatic stress disorder (PTSD). DATA SOURCES: Research-based articles in the medical and psychiatric literature, review articles, and diagnostic criteria. CONCLUSIONS: PTSD is a disorder where psychological and physiological reactions are closely related. Populations at risk include survivors of motor vehicle accidents, natural and man-made disasters, torture, and childhood sexual abuse, as well as combat veterans. IMPLICATIONS FOR PRACTICE: Patients with a history of recent trauma should be targeted for early screening and intervention. Effective treatment of PTSD is a three-stage process consisting of stabilization of symptoms, processing of traumatic perceptions, and integration of trauma into the patient's world- and self-view. PMID- 11930389 TI - Reducing needle-stick injuries with the use of 2-octyl cyanoacrylates for laceration repair. AB - PURPOSE: To describe the effectiveness of 2-octyl cyanoacrylates (2-OCA) for repair of lacerations, reduction of needle-stick injuries to health care providers, patient satisfaction, and overall cost reduction. DATA SOURCES: A descriptive study of typical lacerations (N = 38) presenting to a suburban urgent care center. CONCLUSIONS: Wound approximation was complete (92.1%); with no erythema, drainage, or warmth (100%) and patients were satisfied (87%) and preferred 2-OCA (91%). No needle stick injuries occurred (100%) to providers or ancillary staff. IMPLICATIONS FOR PRACTICE: 2-Octyl cyanoacrylates tissue adhesive effectively closes and heals wounds without infection, pain, or erythema. Health care providers can prevent the risk of blood and body fluid exposure from needle-stick injuries through the use of 2-OCA tissue adhesive. PMID- 11930390 TI - Proper use of skin adhesives for wound repair. AB - PURPOSE: To describe the proper application of tissue adhesive for wound closure in the clinical setting. DATA SOURCES: Product information and clinical experience of the author. CONCLUSIONS: A thin film of adhesive that extends at least one-half centimeter on each side of the opposed wound edges provides the best coverage. Avoid seepage into the wound and runoff onto surrounding tissue to prevent complications. IMPLICATIONS FOR PRACTICE: Proper application assures the best results. Patient information is provided to assure adequate healing. PMID- 11930391 TI - A review of current weight management: research and recommendations. AB - PURPOSE: To review current research and recommendations on weight loss and weight control and provide suggestions for health care providers who furnish weight management counseling. DATA SOURCES: Scientific publications, clinical guidelines, and government sources. CONCLUSIONS: Research reaffirms the long-held understanding that weight loss can be accomplished only through a reduction in the number of calories consumed and an increase in exercise. Weight maintenance requires life-long behavioral change combining moderate exercise, lower fat intake, increased fruit and vegetable consumption, as well as social support. Fad diets and medications are not the answer to long-term weight maintenance. IMPLICATIONS FOR PRACTICE: The essential components of a weight loss or weight management program include: calorie reduction of 300-500 calories per day, appropriate exercise, variety in food choices, increased consumption of grains, fruits, and vegetables, and reduction of fat to no more than 30% of daily calories. Clients should be referred to dietitian and exercise consultants as needed. PMID- 11930392 TI - Promoting health in older adults: a four-year analysis. AB - PURPOSE: The purpose of this study was to consider the influence of selected health promotion and disease prevention interventions in elderly residents of a continuing care retirement community (CCRC) over a four-year period by comparing actual health promotion practices of the residents. DATA SOURCES: Original research using a descriptive design, face-to-face interviews of residents (N = 176-200), chart reviews, and administration of a mini-mental state exam (MMSE) and health survey administered annually. CONCLUSIONS: In each year the mean age of the residents was at least 85, the majority were female, Caucasian, and unmarried. With the exception of checking stools for occult blood, there was a statistically significant change in all health promotion behaviors over the four year period. The most significant change was in the area of exercise behavior, which increased from 24% of the residents participating in regular exercise in year one to 61% by year four. IMPLICATIONS FOR PRACTICE: The purpose of health promotion and disease prevention in older adults is to reduce the potential years of life lost in premature mortality and ensure better quality of remaining life. In addition to regularly scheduled interventions (group education, on-site administration of pneumonia and flu vaccines, on-site exercise room and walking group), individualized counseling regarding the pros and cons of health-promotion activities was provided to help residents make an educated decision about engaging in these activities. These interventions can be used to help facilitate participation in health promotion activities as appropriate and desired for each older adult. PMID- 11930393 TI - Evaluating and applying clinical practice guidelines. AB - Clinical practice guidelines (CPG) hold great potential for providing a summary of large volumes of clinical evidence and a related set of practical recommendations. Nurse practitioners should become aware of the range of available CPGs and methods by which they can be evaluated for use. Appropriate evaluation of CPGs should include their overall reliability and validity, as well as their applicability in specific situations. This article provides an overview of an appropriate evaluation method and serves as an introduction to future columns presenting individual CPGs. PMID- 11930394 TI - A comparison of perspectives on breastfeeding between two generations of black American women. AB - PURPOSE: To determine differences in breastfeeding perspectives between two generations of black American women with and without access to governmental food assistance programs (i.e., WIC). DATA SOURCES: Descriptive, comparative study of a convenience sample of 118 black American women in their childbearing years and beyond conducted in a primary rural health care clinic serving an indigent population. CONCLUSIONS: A significant difference was found between breastfeeding perceptions and rate among younger black American women on WIC program and their mothers without access to these programs. The availability of free formula through WIC programs has partially influenced the rate of breastfeeding among the young black American women. The other significant influencing factor was public embarrassment at breastfeeding. IMPLICATIONS FOR PRACTICE: An extensive educational campaign is needed in order to influence the public's perceptions of breastfeeding as an embarrassment. The success of programs such as WIC must be measured beyond the first six months of an infant's life. PMID- 11930395 TI - Infectious diseases of refugees and immigrants. PMID- 11930396 TI - Viral upper respiratory infections. AB - Clinical practice guidelines (CPG) hold great potential for providing a summary of large volumes of clinical evidence and a related set of practical recommendations. Nurse practitioners should become aware of the range of available CPGs and methods by which they can be evaluated for use. Appropriate evaluation of CPGs should include their overall reliability and validity, as well as their applicability in specific situations. This article is part of an ongoing series presenting individual CPGs. PMID- 11930397 TI - Ascariasis. PMID- 11930398 TI - An analysis of research and clinical practice in neonatal pain management. AB - PURPOSE: To review the literature on neonatal pain management with a focus on historical misconceptions, ethical issues, barriers to practice, the role of the advanced practice nurse (APN), and suggested improvements. DATA SOURCES: Selected research and review articles in nursing and medical literature. CONCLUSIONS: Health care professionals agree that neonates experience pain and in turn deserve effective treatment. Research is controversial regarding the extent of pain management necessary in relation to short-term benefits and long-term consequences. Ethical issues arise when research supporting pain management is not consistently utilized in nursing practice. IMPLICATIONS FOR PRACTICE: Protocols and standardized pain management strategies have demonstrated a beneficial effect on overall patient outcomes. The APN is identified as being in the optimal position to facilitate enhanced neonatal pain management through research, education, and direct clinical care. PMID- 11930399 TI - A motivational intervention to improve adherence to treatment of chronic disease. AB - PURPOSE: To describe an approach to intervention, based on Motivational Interviewing (MI) and the Transtheoretical Model (TTM) of Change that allows the health care provider to support medication adherence in a client-centered fashion. DATA SOURCES: Review of selected research-based and theoretical articles on MI, TTM, and medication compliance issues. CONCLUSIONS: Adherence is a complex phenomenon that requires a personalized intervention. The response must focus on clients' readiness to adhere, their sense of self-efficacy in taking medicines in all circumstances, and their personal pros and cons related to adherence. IMPLICATIONS FOR PRACTICE: Adherence to complex medication regimens is important to long-term treatment of chronic diseases, such as HIV disease. Adherence in HIV is particularly critical to adequately suppress viral replication as well as to prevent opportunistic infections. PMID- 11930400 TI - Cardioprotective benefits of hormone replacement therapy. AB - PURPOSE: To examine the effect of hormone replacement therapy (HRT) on adverse cardiac events in postmenopausal women. DATA SOURCES: A systematic review of all available scientific literature (513 articles) on the cardioprotective benefits of HRT, including research reports, meta-analyses, and commentaries. CONCLUSIONS: Epidemiological studies have consistently suggested that HRT administered to postmenopausal women significantly lowers the risks of cardiovascular events. However, the claimed benefits might have been exaggerated. The results of the only large randomized clinical trial did not show similar benefits as those reported from observational studies. Research findings about cardiovascular protection of HRT remain inconclusive. IMPLICATIONS FOR PRACTICE: While waiting for more definitive data, it is appropriate to consider other options that have been proven safe and effective in the prevention of cardiovascular disease. PMID- 11930402 TI - Hypertension outcomes in an urban nurse-managed center. AB - PURPOSE: To assess the quality of hypertension (HTN) care in an urban nurse managed center (NMC) by chart audits of insured and uninsured (N = 52) African Americans who were managed by nurse practitioners. DATA SOURCES: A chart audit form was developed by the authors that merged Health Plan Employer Data and Information Set (HEDIS) criteria with the Joint National Committee on Detection, Evaluation, and Treatment of High Blood Pressure (JNC VI) criteria. CONCLUSIONS: There was near comparable hypertension control among the two groups. No difference was found in systolic blood pressure (BP) control; however, the uninsured group had a slightly greater average diastolic BP compared with the insured group. There was no significant difference in the number of HTN medications or the number of risk factors. A significant difference was found in the number of NP visits per year between the two groups; the uninsured group averaged 3.2 more visits per year. IMPLICATIONS FOR PRACTICE: Nurse practitioners in this NMC were able to manage HTN in a high-risk population despite a lack of insurance coverage for anti-hypertensive prescriptions. The finding that the uninsured group had more clinic visits per year than the insured group is significant in that it increases the cost of providing care for these patients and the health care system. The patient cost in time, transportation and burden needs further assessment. PMID- 11930401 TI - Health care provider communicator style and patient comprehension of oral contraceptive use. AB - PURPOSE: To explore reasons for the incorrect usage of oral contraceptive pills (OCPs) by examining the relationship between patients' abilities to comprehend and/or recall information presented to them by nurse practitioners (NPs) and the communicator style of their NP. DATA SOURCES: A convenience sample of 46 OCP users completed an instrument designed to test their comprehension of OCP use and an instrument designed to measure the communicator style of their NP. CONCLUSIONS: A multiple stepwise regression revealed that attentive and friendly communicator styles were positive predictors and the communicator image and dramatic styles were negative predictors of comprehension. PRACTICE IMPLICATIONS: Unwanted pregnancies that result from non-compliance with OCP regimens can have significant social and financial effects. A lack of understanding of proper OCP use may depend on the style of communication a NP uses to convey crucial information regarding the regimen. PMID- 11930403 TI - The role of computer-assisted simulation in nurse practitioner education. AB - PURPOSE: To develop a better understanding of the role of computer-assisted simulations (CAS) in nurse practitioner (NP) education by observing 4 pairs of students complete a CAS case. DATA SOURCES: Qualitative observational study of 8 students performing in a computer laboratory and a series of post-observation interviews. Observations were recorded, refined, coded, and quantified based on the theoretical framework of ecological psychology. CONCLUSIONS: Each pair of students established their own "personalities" for the completion of the task. Misinterpretation of information was common and the absence of a live patient interaction affected the reasoning process of the students. The students demonstrated the ability to develop a perspective on the case based on previous nursing experience. Students generally obtained adequate data on which to base differential/final diagnoses. Observations provided information regarding strengths and weaknesses of students and methods used to solve scenarios. IMPLICATIONS FOR PRACTICE: All students agreed that the CAS scenarios were realistic, useful learning experiences, and that the knowledge gained would be transferable to real clinical situations. Such experiences provide useful experience with terminology, sequencing of the examination, and decision-making. PMID- 11930404 TI - [Smoking and dentistry]. PMID- 11930405 TI - [Health damage caused by smoking]. AB - This paper describes the detrimental effects of smoking. More than 30% of the male and female Dutch population still smokes cigarettes. This figure compares unfavorably in international perspective. The smoking habit is responsible for approximately one third of all deaths from cardiovascular, pulmonary and malignant disease. Cigarette smokers face a 50% chance of dying as a result of a smoking related disease. The morbidity associated with smoking is also very substantial. Subjects who discontinue smoking before being struck by disease, and patients who quit the habit once early symptoms develop, have a more favorable outcome. For these reasons, and also because most patients esteem the opinion of their doctor, smoking behavior should be discussed during regular patient doctor contacts, and followed by the advice to quit smoking. PMID- 11930406 TI - [Smoking and pregnancy]. AB - Smoking is disadvantageous for health in every respect. This is a fortiori the case during pregnancy. Pregnant women who smoke burden their unborn child with a mixture of hundreds of tobacco combustion products, dozens of which have been proved to be harmful, apart from the negative influence of the smoke induced blockade of the haemoglobin by carbon monoxide. Since the embryonic and foetal tissues which are formed in a short period, are particularly vulnerable, many detrimental effects can be observed. The fertility rate of women who smoke is reduced and their risk of abortion, miscarriage and preterm delivery is increased. Babies born to women who smoked during pregnancy have lower birth weights and are less developed. They run an increased risk of many disorders including asthma and even cancer. Their cancer risk seems even to be increased in adulthood. Smoking fathers also damage the development of their unborn child. PMID- 11930407 TI - [Smoking and lung cancer]. AB - Since fifty years it is clear now that smoking of tobacco products is responsible for the lung cancer epidemic that is currently in progress worldwide. Although in the Western world a small decrease of lung cancer in males is found, the number of female patients is steadily increasing. Changes in tobacco production have resulted in exposition of smokers to other carcinogens. This is probably the cause of the change in the histological pattern with an increase of adenocarcinoma and stabilisation of squamous cell lung cancer. Despite the bad prognosis there is some hope that with improvement of early detection methods more patients can be cured. However, for a real change it is necessary to discourage smoking by all means. PMID- 11930408 TI - [Smoking and oral diseases]. AB - There are various oral diseases related to tobacco habits. In the majority of these diseases there is a dose-response relationship. This is particularly true for premalignant lesions, such as leukoplakia, and squamous cell carcinoma. Patients should be informed about the deleterious effects of tobacco on the oral tissues. At the same time the patient should know that cessation of the tobacco habits does eliminate the increased risk of developing oral precancer and cancer within a reasonable period of time. PMID- 11930409 TI - [Smoking as a risk factor for periodontitis]. AB - Smokers have a higher susceptibility for periodontitis than non-smokers. Smoking is associated with more loss of attachment, progressive periodontal breakdown and also with a poor response to periodontal treatment. This is caused by the direct effect of nicotine on several factors that play a bacterial attack. These factors include the gingival vascularization and cellular defense mechanisms. The periodontal status of former smokers is intermediate to that of those who have never smoked and current smokers. Smoking cessation seems to have a beneficial effect on periodontal health. PMID- 11930410 TI - [How effective are smoking cessation campaigns?]. AB - An important risk factor, single as well as combined with other factors, for the development of cancer is tobacco smoking. In the Netherlands 37% of the adult men smoke, and 30% of the women. It is estimated that 20% of the now living population will eventually die from the consequences of smoking. Besides stimulating youngsters not to take up smoking, helping smokers to stop is preventative for the evolution of cancer. However, how effective are the attempts that are currently accomplished in this field? In this contribution, the quit rates that the different methods produce will be described, over and above the 5% of smokers who yearly succeed in quitting on their own. These rates vary from 5% among the general population resulting from a mass media led campaign. Finally, some critical comments will be given on governmental policies. PMID- 11930411 TI - [Are dentists willing to advise their patients to quit smoking?]. AB - In order to know whether dentists are prepared to advise their patients to stop smoking, a representative study was performed in several EU-countries. From the Dutch results, it may be concluded that dentists in the Netherlands are aware of the harmful effects of tobacco use. They are prepared to pay more attention in their dental practice to these harmful effects for general health and for oral health in particular. The majority is willing to participate in a smoking cessation campaign, provided there is more information about 'why' and 'how'. PMID- 11930412 TI - [A misleading cheek lesion]. PMID- 11930413 TI - Peer review: how it should be done. PMID- 11930414 TI - Weight management practices among primary care providers. AB - This pilot study examined how primary care providers manage patients with weight problems, an important component of primary care. A convenience sample of 17 nurse practitioners and 15 physicians were surveyed about assessments and interventions used in practice for weight management along with perceived barriers to providing effective weight management. Practice patterns between gender, profession and practice setting of the nurse practitioners were compared. PMID- 11930415 TI - Development of a student nurse practitioner computerized clinical log. AB - A computerized clinical log was developed and revised over a period of three semesters by the family nurse practitioner students at the University of Texas Health Science Center in San Antonio, Texas. Students entered data on a disk formatted in Microsoft Access, and results were analyzed by the Statistical Packages for the Social Sciences (SPSS). More than 4,000 patient encounters were documented during three semesters. The development and refinement of the computerized log are described. Results are presented and the implications of the database for educational, curricular and employment purposes are discussed. PMID- 11930416 TI - Coping strategies of rural families of critically ill patients. AB - This study explored the coping strategies of families of critically ill patients in a rural Southern Appalachian setting. A convenience sample of 30 family members of 22 critically ill patients in two rural hospitals completed the Jaloweic Coping Scale. The five most frequently used coping methods were helping, thinking positively, worrying about the problem, trying to find out more about the problem and trying to handle things one step at a time. The five most effective coping strategies were talking the problem over with friends, praying, thinking about the good things in life, trying to handle things one step at a time and trying to see the good side of the situation. Findings contradicted many of the more "negative" descriptions of Appalachian people in the literature. Similarities outweighed differences when comparing the coping styles of rural and urban populations. Findings suggest that coping strategies must be considered for positive outcomes in the delivery of care to such a rural population. PMID- 11930417 TI - Diagnosing appendicitis across the life span. AB - Arriving at a definitive diagnosis of acute appendicitis (AA) requires an understanding of the various presenting signs and symptoms of all ages and genders across the life span. Primary care providers must also be aware of the pertinent laboratory tests and imaging procedures that will assist in providing useful information toward making an accurate diagnosis of AA. Utilizing an in depth review of the literature, this article delineates the presenting signs and symptoms of AA from children to adults to elders. It also delineates which laboratory and imaging techniques should be used for each age group as effective adjuncts in diagnosing AA. PMID- 11930418 TI - The A, B, C's of bone building in adolescence. AB - The majority of treatment for osteoporosis has focused on secondary and tertiary strategies that attempt to preserve acquired adult bone mass. However, the occurrence of osteoporosis and its related physical and economic sequelae continue to escalate, resulting in devastating morbidity and mortality in later adulthood because of bone fractures. Mounting evidence continues to support primary intervention strategies that target adolescents during skeletal maturation as effective in building bone mass reserves with the power to resist the threat of osteoporotic fractures. Nurse practitioners need to exercise their expertise as primary care providers by teaching adolescents and their families the A, B, Cs of bone building with calcium, vitamin D and exercise. PMID- 11930419 TI - Evaluation of a single seizure: guidelines for advance practice nurses. AB - Deciding whether or not to treat a client after a single seizure can be challenging for the clinician. The risk of seizure recurrence is greatest in the first six months after the initial episode. The decision to treat a single seizure should be based on diagnostic and clinical findings that assess the client's risk for recurrence. When making treatment decisions, the clinician must consider the benefits and risks of single-seizure therapy. Careful consideration should be given to differential diagnosis, factors that induce symptomatic seizures and risk factors for epilepsy. PMID- 11930420 TI - Creating a "health connection" between STD services and primary care. AB - Women's health nurse practitioners may find their practice includes men who present for treatment of sexually transmitted diseases (STD). In seven months, the women's health nurse practitioner at Temple Health Connection treated 70 African-American males for STDs. Seventeen males (24%) were diagnosed with hypertension. Nine of these patients returned for follow-up care with a primary care adult nurse practitioner. Our experience shows it is important to combine STD treatment with accessible primary health care. PMID- 11930421 TI - Urinary tract infections. AB - Urinary tract infections are some of the most commonly treated bacterial infections in adults. Factors contributing to development of UTIs include personal habits, sexual habits, altered immune status, presence of diabetes, lack of estrogen and use of spermacides. Untreated or persistant UTIs may cause other more serious complications such as kidney damage or prostate infections. Symptoms that persist after appropriate treatment need further evaluation. As with any disease, appropriate patient education is necessary to help the client understand the process and follow an effective treatment plan. PMID- 11930422 TI - [Antibiotic resistance in the subgingival microflora in patients with adult periodontitis. A comparative survey between Spain and the Netherlands]. AB - The widespread use of antibiotics for treatment of bacterial infections has lead to the emergence of resistant human pathogens. Great differences have been documented between European countries in the use of systemic antibiotics. In parallel, significant differences in levels of resistant pathogens have been documented. In order to investigate whether differences in antibiotic use influence the level of antimicrobial resistance of the subgingival microflora, microorganisms from the subgingival plaque of untreated patients with adult periodontitis in The Netherlands (n = 30) and Spain (n = 31) were compared. Blood agar plates containing breakpoint concentrations of penicillin, amoxicillin, amoxicillin and clavulanate, metronidazole, erythromycin, azithromycin, clindamycin and tetracycline were used to determine the proportion of bacteria from the subgingival plaque that was resistant to these antibiotics. In the Spanish patients, statistically significant higher mean levels of resistance were found for penicillin, amoxicillin, metronidazole, clindamycin and tetracycline. The mean number of different bacterial species growing on the selective plates was higher in the Spanish patients, as was the percentage of resistant strains of most periodontal pathogens. A striking difference was observed in the frequency of occurrence of tetracycline-resistant periodontal pathogens. In Spain, 5 patients had > 3 tetracycline resistant periodontal pathogens, whereas this was not observed in any of the Dutch patients. It is concluded that the widespread use of antibiotics in Spain is reflected in the level of resistance of the subgingival microflora of adult patients with periodontitis. PMID- 11930423 TI - [Formaldehyde in dental materials]. AB - The use of formaldehyde in dentistry has been discussed for years. This because of the possible systemic effects of its use. This paper addresses the possible systemic effects of the use of formaldehyde and the question in which dental products formaldehyde is a component. The indications for the use of formaldehyde as its alternatives are listed. The conclusion is that for nearly all dental indications for the use of formaldehyde good alternatives are available. PMID- 11930424 TI - [Role of inheritability of tooth form, tooth malformation and tooth position]. AB - A common technique to divide the influence of heredity and environment on certain characteristics, or pathologies is the one that uses twins. There are more or less complex techniques to carry out twin research, from which the most simple procedure consists of determining the amount of concordance of certain characteristics in monozygotic (MZ) and dizygotic (DZ) twins. In individuals who develop from one oocyte, like MZ twins, one would expect a correlation of 1.00 (or 100% concordance) purely from their gene-relation. DZ twins are the result of separate conceptions of the same parents and they thus differ as much from each other as ordinary brothers and sisters. The most recently developed twin methodologies use path analysis and model fitting for the estimations of the heritabilities and environmental influences on certain characteristics. In this article it is tried to picture the genetic an environmental influence on tooth form, tooth position and occlusal characteristics with different genetic techniques. Generally it can be concluded that our genes are of utmost importance for tooth form and tooth malformations, but the environment has a much bigger impact on tooth position and occlusal parameters. PMID- 11930425 TI - [Syndromes 13. Epidermolysis bullosa]. AB - Epidermolysis Bullosa is characterised by blister formation of skin and mucous membranes. Three major varieties of epidermolysis bullosa (EB) are reviewed including their dental and oral aspects: EB simplex, junctional EB and dystrophic EB. Marked oral involvement of the soft and hard tissues can produce potentially devastating alterations in anatomy and function. Oral debilitation is limited primarily to the recessive dystrophic EB type due to soft tissue scarring following blister formation. Microstomia, ankyloglossia and obliteration of the oral vestibule are typical features of dystrophic EB. In other cases enamel hypoplasia and cementum disorders can be present. Epidermolysis bullosa has considerable impact on oral health and dental care. PMID- 11930426 TI - [An acute unilateral hypoglossus nerve palsy]. PMID- 11930427 TI - [Herpes simplex infections]. PMID- 11930429 TI - [Topics in clinical dentistry. Trends in the Dutch dental literature]. AB - Aim of the study was to evaluate a method for selecting topics suitable for developing dental clinical practice guidelines in the Netherlands, based on an analysis of Dutch dental journals. A search for dental clinical topics was conducted by analysing Dutch dental journals, magazines and series over the period 1992-1997. The numbers of publications per topic were plotted against the publication years. The number of publications as well as the value of the slope of the linear regression were considered to be indicators of the importance of a topic. 'Dental implants (indication)' had the highest number of publications, followed by 'orthodontic treatment planning' and 'periodontology (indication)'. The topic 'practice hygiene' showed the highest value of the slope of the linear regression, followed by 'TMJ dysfunction' and 'dental implants (indication)'. With this method, it is feasible to detect changes and tendencies in the Dutch dental literature. It permits a selection of clinically relevant topics over a time span. It was concluded that this method may be very useful in the selection of a topic, but should probably be combined with other methods. PMID- 11930428 TI - [Considerations for dental care during pregnancy]. AB - Traditionally, general dentists have been reluctant to treat pregnant patients due to uncertainty of the risks that might be imposed on both the mother and the fetus. However, preventive, emergency, and routine dental procedures are all suitable during various phases of a pregnancy, with some treatment modifications and initial planning. This article will review some of the physiologic changes and oral pathology associated with pregnancy, and how these alterations affect dental care for the patient. A brief discussion concerning the risks and benefits of radiation, certain medications, and pre-natal fluoride is also given. Additionally, an overview of possible obstetric emergencies that may arise during a general dental appointment is presented with treatment options outlined. PMID- 11930430 TI - [Syndromes 15. Robin sequence]. AB - Robin sequence (RS) is a congenital anomaly that includes micrognathia, U-shaped cleft palate, and upper airway obstruction. More than 80% of children with RS have other malformations as a result of associated congenital anomalies. This report briefly reviews some multidisciplinary aspects, including etiopathology, diagnosis and treatment. PMID- 11930431 TI - [Autotransplantation; not by definition a root canal treatment?]. PMID- 11930432 TI - [Colon carcinoma 2]. PMID- 11930433 TI - [Dental caries prevalence in youth from the Hague. Have the differences in dental health between social groups closed?]. AB - In 1998 a dental survey amongst 6- and 12-year-old schoolchildren in the Hague was carried out. The 1998-survey showed differences in caries experience between SES groups that had increased in the period 1989-1998. The decline in caries experience among 6-year-old children of low SES groups had come to an end after 1983, whereas among 12-year-old children the decrease in mean DMFS values had stopped in the early nineties. However, the survey showed that during 1989-1998, the percentages of caries-free children in medium and high SES groups continued to rise. Of the medium SES 6- and 12-year-old children, In 1998, the percentage of caries-free 6- and 12-year-old children in medium SES group were 79 and 87, respectively. The comparable results in the high SES group were 87 (6-yr-olds) and 93 (12-yr-olds). PMID- 11930434 TI - [Reproducible marginal gap measurements of partial ceramic crowns]. AB - Aim of this research was to test the hypothesis that marginal gap measurements by a digital microscopic image processing computer are reproducible. Light microscopic images of the margins of Artglass partial crowns on stone dies were obtained with a 3CCD color video camera, digitized and processed. Ten measuring points were painted on a 0.5 mm section of the finish line of the preparation. The opposite corresponding measuring points on the margin of the partial crown were painted as well. The distance (marginal gap) between two corresponding points was painted and calculated by the computer. Reproducibility (precision) is reflected in the ability to reproduce the same measurement results in repeated measurements (n = 5). The reproducibility error was expressed as the coefficient of variation (CV) in percent (%). The localization of the corresponding measurement points and the calculation of the marginal gap were performed without errors. For a marginal gap between 15-50 microns the average measurement error was 3% or 0.45-1.5 microns. It was concluded that the measurement of the gap between two points by the digital image processing computer was reproducible. The measurement error was neglectable given that a clinically acceptable marginal gap is 100 microns. PMID- 11930435 TI - [Cutaneous and mucosal reactions to dental materials]. AB - The concentration of substances released by dental materials is usually too low to cause systemic-toxic effects, but there is the potential to induce dermatoses in especially dentists as well as mucositis in mainly dental patients. The cytotoxic irritative and allergic diseases are successively described categorized according to their pathogenesis. The identification of the diseases, which often have a similar appearance, depends on history-taking and patch testing. Non dental sensitization is of diagnostic importance. PMID- 11930436 TI - [Syndromes 14. Rendu-Osler-Weber disease]. AB - The Rendu-Osler-Weber disease is due to an autosomal dominant disease with multiple telangiectasia in skin and mucosa. Recurrent bleeding of the nose is due to telangiectasia of the nasal mucosa. Haemorrhage of the oral mucosa also occurs. Extensive arteriovenous malformations can be present in lungs, liver and brain. Treatment of bleedings in the oral mucosa is easily possible by means of coagulation. Patients with pulmonary arteriovenous malformations should receive prophylactic antibiotic treatment before extractions, removal of third molars and subgingival curettage. PMID- 11930437 TI - [Periapical radiographic abscess with dental implantation]. PMID- 11930438 TI - [Colon carcinoma I]. PMID- 11930439 TI - Patient satisfaction with nurse practitioner service in a rural setting. AB - PURPOSE: To determine the level of patient satisfaction with service provided by nurse practitioners (NP) to rural patients. DATA SOURCES: Original research utilizing the Nurse Practitioner Satisfaction Instrument (NPSI), developed by the researcher and completed by a convenience sample of 93 rural patients. CONCLUSIONS: Overall there was a high level of patient satisfaction with NP service. Significant relationships (p < or = .05) were identified between patient satisfaction and the subject's age, educational level, familiarity with the NP seen, overall health, satisfaction with expectations of service being met, and likelihood to recommend the NP to others. IMPLICATIONS FOR PRACTICE: These findings can be used to support the effectiveness of NP providers and the need for less restriction in reimbursement policies, which in turn, may improve access to care in rural and underserved areas. PMID- 11930440 TI - Repetitive strain injuries in youth. AB - PURPOSE: To review the anatomic and physiologic considerations in repetitive strain injuries (RSI) in youth in relation to normal growth and development. DATA SOURCES: Extensive review of scientific literature, laws relating to youth labor, and the authors' experience. CONCLUSIONS: Early detection of youth at risk for RSIs is crucial for prevention and treatment. Physical maturity and size are more important than chronological age in the safe performance of work tasks or sports. IMPLICATIONS FOR PRACTICE: Interventions that reduce injury in youth include gradually increasing training load, improving flexibility, and strength, improving biomechanics, and correcting malalignment. Assessments should include tests of flexibility, laxity, and muscle symmetry. PMID- 11930441 TI - Development of a local advanced practice nurse coalition. AB - PURPOSE: To describe the experiences of a group of advanced practice nurses (APNs) in a rural area in developing an APN Coalition. DATA SOURCES: Selective review of literature and documents from the newly developed organizational framework for the coalition. CONCLUSION: Professional connections can begin at the local level through the development of local APN coalitions and then expand to involvement in state and national organizations. IMPLICATIONS FOR PRACTICE: Guidelines for establishing the coalition organizational structure are included and can be used as a template by similar evolving APN coalitions. PMID- 11930443 TI - Nurse practitioners and physicians: patients' perceived health and satisfaction with care. AB - The advent of managed care has created changes in the health care environment and nurse practitioners have found a need to evaluate their care. Perceived health and patient satisfaction were measured in a multiethnic sample of 160 clinic patients, ages 18 to 89, in a managed care setting. Results of the Medical Outcomes Study SF-20 and the Nurse Practitioner Satisfaction Instrument indicated no statistically significant difference in perceived health and satisfaction with care, whether the care was given by a nurse practitioner or a primary care physician. The findings warrant further study and may mean that nurse practitioners placed in managed care environments can be expected to perform as effectively as they have in non-managed care environments. PMID- 11930442 TI - Treatment of rheumatoid arthritis: etanercept a recent advance. AB - PURPOSE: To review the specific mechanism of action of biologic response modifiers and help clinicians place these new drugs in the context of established therapies for rheumatoid arthritis (RA). DATA SOURCES: Review of scientific literature, including all the clinical trials for these drugs. CONCLUSIONS: Biologic response modifiers target key inflammatory events involved in joint destruction. They have the potential to revolutionize the management of RA. IMPLICATIONS FOR PRACTICE: Referral to a rheumatologist should be strongly considered as soon as the diagnosis of RA is made. Early referral to a specialist has been associated with improved health status for patients with RA due to early and aggressive therapy with these new agents. PMID- 11930444 TI - Professionalism and the nurse-midwife practitioner: an exploratory study. AB - This article reports the findings of an exploratory study on the level of professionalism among certified-nurse midwives (CNMs). Using a five dimensional scale to measure professionalism, our research examined the level of professionalism among CNMs. It explored the relationship between organizational reward structures, level of education, and professionalism. The results show that CNMs exhibit a high degree of professionalism in the practice of nurse-midwifery. The findings also support the notion of a causal relationship between reward structures, education levels and CNM professionalism. PMID- 11930446 TI - Accessing patient assistance programs to meet clients' medication needs. AB - The high cost of medication is a barrier to patient compliance. Nurse practitioners can access patient assistance programs to assist low-income clients to obtain medications, empowering them to take action to improve their health. At Health Care Access, a primary care clinic serving low-income, uninsured persons, approximately $150,000 worth of medications were obtained through patient assistance programs in 1997, compared to the total clinic budget of $189,000. Processes for obtaining prescription drugs vary between pharmaceutical companies. An understanding of these types of programs provides nurse practitioners access to these resources for their clients. PMID- 11930445 TI - Thyroid disease: a review for primary care. AB - Manifestations of thyroid disease include hyperthyroidism and hypothyroidism. In the past, the diagnosis of thyroid disease has been perplexing due to the multisystemic effects of thyroid hormone and the lack of specificity in laboratory tests. New methods of measuring thyroid function have made screening and diagnosis of thyroid dysfunction both more accurate and cost-effective. Testing for thyroid stimulating hormone (TSH) is now a first-line diagnostic procedure and, in some cases, may be the only test indicated. The general pathophysiology of thyroid, the diagnosis and management of thyroid disease, and current screening recommendations are reviewed here. PMID- 11930447 TI - Pharmacokinetics in children: history, considerations, and applications. AB - Crucial to safe pharmacological intervention in the pediatric population is a thorough understanding of the pharmacokinetic and pharmacodynamic differences in children as opposed to adults. This article discusses these differences and clarifies factors related to medication prescribing and administration techniques. PMID- 11930448 TI - Neurocysticercosis: migration of a parasite. AB - Neurocysticercosis is the most common parasitic infection of the central nervous system (CNS). Infection occurs following oral ingestion of eggs of the porcine tapeworm, Taenia solium. Eggs migrate to distal tissue sites via the bloodstream, and develop into larval cysts. When cyst growth disrupts normal function of the CNS, patients most commonly present with new onset seizures or hydrocephalus. Neuroimaging with computed tomography (CT) scan and magnetic resonance imaging (MRI) is used to diagnose neurocysticercosis. Treatment regimens are based on cyst location and developmental stage, and include supportive care, supportive care combined with pharmacologic therapy, and surgical intervention. PMID- 11930449 TI - International collaboration: initial steps and strategies. AB - The nurse practitioner (NP) movement, which began in the United States in 1965, has caused sweeping changes both in nursing and in health care delivery. The concept of NPs then spread to the United Kingdom (U.K.) and continues to move globally. The international development of NPs has been streamlined through collaborative efforts of the U.S. and the U.K. There have been striking similarities in the evolution of expanded practice roles in both countries. In 1993, NPs from the U.S. and the U.K. came together to support the first international NP conference and, in the process, developed a framework for global international collaboration. This article summarizes the collective knowledge and the strategies for success developed through this collaborative effort. PMID- 11930450 TI - A personal account of a medical mission. AB - The author participated in a medical mission to the Philippines while he was a student in a family nurse practitioner program. This first person account of his two weeks in the Philippines details his impressions of the country, experiences with poverty, and the challenges of working with limited resources. PMID- 11930451 TI - Use of complementary and alternative treatments by individuals with fibromyalgia syndrome. AB - Although muscle pain is the primary complaint of patients with fibromyalgia, there are myriad associated symptoms that cause them to seek healthcare. Some individuals try alternative treatments when conventional medicine does not provide symptom relief. A questionnaire was developed to collect information regarding complementary treatments and their effectiveness. Sixty individuals visited the researcher's web page and completed and submitted an online questionnaire on fibromyalgia. Literature, heat, walking, vitamins, and massage were the interventions tried most frequently. Literature, aromatherapy, support groups, heat, and massage were rated the most effective. PMID- 11930452 TI - An algorithm for monitoring phenytoin therapy. AB - Nurse practitioners need an evidence-based framework for monitoring clients on phenytoin (Dilantin) therapy. A Med-line search of published literature and a synthesis of 10 years of research was performed. The effect of phenytoin within the body is dependent on the amount of serum albumin and the functional status of the major organs in the body. The basic principles of pharmacotherapeutics holds true for phenytoin. Closer investigation of total serum phenytoin and serum albumin ratio by primary care providers is necessary for monitoring therapeutic levels of phenytoin therapy. An algorithm was developed to provide a quick reference for monitoring therapeutic phenytoin levels. PMID- 11930453 TI - Syringomyelia: an expanding problem. AB - Syringomyelia, a central cavitation of the spinal cord, was once thought to be a rare disorder. Associated with congenital malformations as well as trauma and tumors, a syringomyelia can cause a variety of sensory complaints, including parasthesias, changes in temperature recognition, and pain. Other symptoms may range from gastrointestinal problems to shoulder arthropathy. Left unchecked, a syringomyelia can expand and lead to permanent disability. The nurse practitioner should be suspicious of the presence of a syrinx in patients who have otherwise unexplained sensory deficits that follow cervical or thoracic dermatomes. PMID- 11930454 TI - Minors' rights to refuse medical treatment requested by their parents: remaining issues. AB - Nurse practitioners are regularly faced with ethical and legal dilemmas when providing care to minors. Laws may not provide clear direction; there may even be conflicting precedents regarding the status of minors, particularly with regard to the juvenile justice system. This article reviews the status of minors' rights with regard to refusing or consenting to medical tests or treatments. Three cases from one author's (DPG) practice illustrate the issues involved. PMID- 11930455 TI - The challenge of evaluating fatigue. AB - Fatigue, though a common complaint, presents a challenge to nurse practitioners who must evaluate its significance in the primary care setting. Fatigue is described as a multidimensional phenomenon and is classified as physiologic, acute, or chronic. A distinction is made between chronic fatigue and chronic fatigue syndrome (CFS). The use of the Piper Fatigue Scale is suggested in order to objectively quantify this symptom in clinical practice. In this article, the Habit and Lifestyle Form for the Evaluation of Fatigue is presented as a way to assist the provider in considering simple and obvious causes of fatigue that may otherwise be overlooked. Finally, an algorithm for the evaluation of fatigue is presented. PMID- 11930456 TI - Winding roads and faded signs: ethical decision making in a postmodern world. AB - We are living in an era, sometimes referred to as "postmodern," exemplified by complex change related to vast increases in information and technology and exposure to diverse people and ideas. Society as a whole is experiencing dissonance in solving ethical dilemmas, and nurses' ethical dilemmas are never far removed from the social context in which nurses practice. This article explores aspects of postmodernism that complicate ethical decision making. It is hoped that this discussion may aid nurses in understanding how world values, especially those of postmodernism, complicate ethical decision making in health care. Suggestions melding aspects of the postmodern with traditional approaches to ethical decision making are presented. PMID- 11930457 TI - The "right" to a caring relationship: the law and ethic of care. AB - The ethic of care is often described as the ethical basis for nursing. While ethics and legal issues are related, it is unclear if perinatal nurses apply this theoretical relationship to clinical practice. There is evidence that medical malpractice litigation may be motivated by dissatisfaction with the caring relationship. A patient's moral "right" to a caring relationship, barriers in clinical practice to caring relationships, and legal remedy for perceived violation of the right to a caring relationship are discussed. Nurses must promote sound caring relationships and articulate to others the impact on perinatal outcomes and legal liability. PMID- 11930458 TI - Ethical issues for nurses providing perinatal care in community settings. AB - Ethical issues in perinatal nursing are complex in that two patients--mother and fetus--are considered. This work considers six areas of potential ethical conflict: conflict between the mother and fetus, informed consent, confidentiality, cultural conflicts, conflicts associated with managed care, and conflicts in childbirth education. Ethical principles of autonomy, beneficence, and justice are included. Strategies for resolving ethical conflicts in community practice settings are suggested. PMID- 11930459 TI - Moral distress in perinatal nursing. AB - This work focuses on an emerging concept in nursing ethics--moral distress. Unlike an ethical dilemma where one does not know the right thing to do, in contrast, moral distress occurs when one knows the right thing to do but is constrained by the institution or one's coworkers in doing it. Moral distress is a process that involves recognition that a decision is difficult to act upon; the experience of emotional distress inherent in that situation; choosing strategies; and then acting. Whistle blowing and collaborative practice models both provide strategies for moral action in moral distress situations. Less dramatic and quieter forms of moral action will also be examined. Finally, developing the strength to move from moral distress to moral action will be discussed, as well as research implications for this emerging ethical issue. PMID- 11930461 TI - Contextual dynamics of ethical decision making in the NICU. AB - The manner in which the contextual dynamics of the neonatal intensive care unit (NICU) culture influence decisions around withholding and withdrawing treatment for very low birth weight infants is examined based on sociological studies of the NICU culture. The influence of these dynamics on nurse and parent participation in treatment decisions is discussed. Steps toward increasing nurses' role in decision making and ability to empower the participation of parents include (a) using an understanding of the dynamics of the individual NICU to establish a collaborative team culture, and (b) demonstrating that data obtained through relationship with the infant make an important and valid contribution to ethical decision making. PMID- 11930460 TI - Ethical issues in the delivery room: resuscitation of extremely low birth weight infants. AB - The neonatal team attending high-risk deliveries is often faced with difficult ethical decisions concerning aggressive cardiopulmonary resuscitation for infants of 22-24 weeks gestational age (GA). These decisions are often based on the ethical principles of beneficence, nonmaleficence, justice, futility, autonomy, quality of life, and best interests and legal rights of the infant. Three delivery room recommendations are discussed along with their legal and ethical considerations. The perinatal and neonatal intensive care nurse must understand the moral, ethical, legal, and professional responsibilities and their effect on her/his own judgment decisions. These, in turn, will affect the infants, their parents, and other colleagues. PMID- 11930462 TI - Life support decisions involving imperiled infants. AB - The birth of a baby can be a joyous event. When an infant requires neonatal intensive care, concern may overshadow joy. This work reports a descriptive, qualitative study that explored parents' perception of their experiences in the neonatal intensive care unit (NICU) when faced with the dilemma of withholding and/or withdrawing treatment from their infants. Data from open-ended interviews revealed significant themes. Clear, accurate, and timely exchange of information helped parents grasp their situation and feel involved in the decision. When health care providers demonstrated to parents that they cared, it helped promote trust between providers and parents. Trust promoted parents' confidence in information received and ultimately in the decision reached. Thus, parents were focused on relationships with health care providers, not on ethics. PMID- 11930463 TI - Attitudes toward nurse practitioners: influence of gender, age, ethnicity, education and income. AB - Survey research was undertaken to measure relationships between gender, age, ethnicity, education, income level, and an individual's attitude toward using a nurse practitioner (NP) for health care. Pender's Health Promotion Model provided the theoretical basis for the research initiative and instrument design. Following initial pilot work, 238 individuals were surveyed. While no significant differences on the basis of gender and race were found, high school graduates demonstrated significantly more positive attitudes toward NPs than non-high school graduates, and older subjects and those with lower incomes were less positively inclined to use NP services. These findings have implications for the marketing of NP services, NP education, and public education, and should be used as a basis for additional research in this area. PMID- 11930464 TI - Increasing the knowledge base of asthmatics and their families through asthma clubs along the southwest border. AB - This study focused on behavior modification and enhancement of knowledge concerning home asthma management. The intervention focused on asthma awareness, severity assessment, medication use, and development of management plans while building a support group. The quasi-experimental design utilized various tools to evaluate knowledge, behavior modification, and self-care management. A 25% increase in knowledge and a 13% decrease in emergency department visits was identified over the 12-month period of the study. PMID- 11930465 TI - The use of low molecular weight heparin in the initial management of patients with deep vein thrombosis. AB - Low molecular weight heparins (LMWH) have been found to be a safe and effective treatment for the prevention of venous thromboembolism. Recent clinical trials have further investigated the use of this class of drugs in the initial treatment of venous thromboembolism. The interest surrounding this new group of anticoagulants is sparked by the reduction in intensive resources required to safely administer this medication in comparison to the traditional treatment with heparin. Low molecular weight heparins do not require laboratory monitoring and can be given once or twice a day by subcutaneous injection, thereby allowing safe administration in the home. The potential cost savings incurred is timely in light of cost containment needs surrounding the current health care climate. PMID- 11930467 TI - Integration of spirituality into health care practice by nurse practitioners. AB - Although conceptualizations of mind, body, and spirit are ancient, spiritual aspects have not been readily integrated into health care. Western medicine's mechanistic model, with its "fix-broken-parts" paradigm, focuses on the physical body, with occasional consideration given to emotional and mental aspects. One's view of self in relation to a Supreme Being, and one's existence and purpose for life is central to health at all levels, e.g., spiritual, physical, emotional, and cognitive. The purpose of this article is to describe the role that nurse practitioners can and should take with patients and their families in integrating spirituality into health care practice. PMID- 11930466 TI - Hereditary hemochromatosis: should clinicians screen for this common disease? AB - Hereditary hemochromatosis is a serious condition of abnormal iron metabolism that is often missed until complications involving multiple organ systems occurs. It is an autosomal recessive condition primarily affecting Caucasians of Western European origin. This article discusses the pathophysiology, diagnosis, and management of this condition. Questions related to population screening are examined and recommendations made for a cost-effective screening program. PMID- 11930468 TI - Antimicrobial resistance. AB - Antimicrobial resistance is not a new concept. For over half a century, health care providers have been faced with this problem. The overuse and misuse of antimicrobial therapy by health care providers has contributed largely to the problem, but several other factors have also been associated with antimicrobial resistance. This article reviews current literature regarding antimicrobial resistance in an effort to educate health care providers to make judicious decisions in the treatment of bacterial infections and stem the rise of antibiotic resistance by carefully scrutinizing prescribing practices. Contributing factors to antimicrobial resistance and recommendations for the control of antimicrobial resistance will be reviewed. Treatment recommendations for common health ailments (i.e., acute otitis media, rhinitis, sinusitis, and pharyngitis) are provided. PMID- 11930469 TI - Evaluating acute musculoskeletal complaints. AB - Clinical practice guidelines (CPG) are now widely available summarizing large amounts of scientific evidence and providing specific recommendations for the evaluation, diagnosis, and management of varied health problems. In order to take advantage of the available CPGs in clinical practice, providers must be aware of their existence and be able to critique them for scientific merit and relevance to specific clinical settings. This ongoing series is designed to provide a brief review of a different CPG each month. This month, the CPG reviewed addresses the evaluation of an extremely common set of complaints: musculoskeletal symptoms. The document emphasizes the importance of a thorough history and physical for these presentations and indicates when specific diagnostic studies and/or consultations are warranted. PMID- 11930470 TI - Improving compliance in your dyslipidemic patient: an evidence-based approach. AB - PURPOSE: To synthesize the evidence from landmark clinical studies of lipid lowering pharmacotherapy and nurse management of hyperlipidemia, discuss issues related to nonadherence, and proposes strategies for achieving long-term cholesterol control. DATA SOURCES: All publications of lipid-lowering clinical trials related to pharmacotherapy for dyslipidemias were accessed from a thorough Medline Search and reviewed by two nurse experts. CONCLUSIONS: Randomized controlled studies provide compelling evidence that reduction of blood cholesterol with pharmacotherapy reduces both first and subsequent coronary events. Nonetheless, inadequate provider and patient adherence to guidelines for lipid lowering remains prevalent. Studies show that nurses provide safe and effective care for patients with abnormal lipids. IMPLICATIONS: This article assists nurse practitioners in playing an active role in the implementation of the National Cholesterol Education Program Adult Treatment Panel III Report to be released in Spring 2001, where a strong emphasis will be placed on multidisciplinary approaches and adherence. PMID- 11930471 TI - Hepatic hydrothorax: a retrospective case study. AB - PURPOSE: To present the pathophysiology, differential diagnoses, assessment techniques, and treatment options for hepatic hydrothorax. DATA SOURCES: A case study is presented with supporting material from current medical literature. CONCLUSIONS: Hepatic hydrothorax is a pleural effusion caused by the flow of ascitic fluid into the pleural space through an actual defect in the diaphragm. Successful outcomes depend on early detection and timely referral of often-subtle lung involvement. IMPLICATIONS FOR PRACTICE: Although incidence is reported to be as high as 12% in cirrhotic patients, standard medical references attach little importance to pulmonary risks in this population. Hepatic hydrothorax should always be considered in the cirrhotic patient with a pleural effusion. PMID- 11930472 TI - Individualizing care of hepatitis B in older adults and Asians. AB - PURPOSE: To review the pathophysiology of hepatitis B virus (HBV) as it relates to special groups such as older adults and Asian populations and to stratify preventive services relative to these particular populations. DATA SOURCES: Selected scientific literature and practice guidelines. CONCLUSIONS: The HBV is one of the most common causes of viral hepatitis. Acute infection is mostly self limiting with nonspecific symptoms. Chronic infection is characterized by persistent serum levels of Hepatitis B surface antigen (HBsAg) for more than six months. IMPLICATIONS FOR PRACTICE: Chronic HBV is a major risk factor for hepatocellular carcinoma. Special populations, such as older adults with decreased immune response and Asians from places with high endemic HBV infection, are at particularly high risk for progression of HBV infection to chronic states and serious liver damage. PMID- 11930473 TI - Lyme disease. AB - PURPOSE: To review the presentation, treatment, and prevention of Lyme disease in primary care. DATA SOURCES: Selected articles from the scientific literature and the Centers for Disease Control. CONCLUSIONS: Lyme disease is a tick-borne, multisystem inflammatory disease with worldwide distribution caused by the spirochete Borrelia burgdorferi. Lyme disease is initially characterized by a spreading, annular erythema migrans skin rash and can disseminate to the musculoskeletal, neurologic, or cardiovascular system. Stages of the disease present with varying clinical manifestations. IMPLICATIONS FOR PRACTICE: Treatment of Lyme disease with appropriate antimicrobial therapy should be prompt in order to avoid debilitating outcomes. Awareness of the many presentations of Lyme disease in endemic areas may facilitate a timely diagnosis and appropriate treatment. PMID- 11930475 TI - Primary care providers' views of precepting nurse practitioner students. AB - PURPOSE: To explore perceived barriers to and facilitators of precepting as viewed by experienced community-based preceptors. DATA SOURCES: Twenty-four nurse practitioner and physician preceptors participated in a 20-minute telephone interview, using a semi-structured format. The interviews were audio-taped and transcribed. Constant comparative analysis was used to organize the preceptors' responses into thematic categories. CONCLUSIONS: Overall, most preceptors were satisfied with the precepting experience; however, many preceptors cited increasing difficulty with teaching students in primary care sites due various organizational constraints such as the imperative to increase productivity. Good communication with school of nursing faculty was cited as the key factor to continued willingness to precept. IMPLICATIONS FOR PRACTICE: Understanding factors that are important for preceptors continuing in their clinical teaching role is important for schools of nursing, especially given the increasing time limitations that many providers face due to social and economic changes in health care delivery. PMID- 11930474 TI - The effect of age, gender, risk level and glycosylated hemoglobin in predicting foot amputation in HMO patients with diabetes. AB - PURPOSE: To determine if age, gender, risk level and glucosylated hemoglobin could predict foot amputations in HMO patients with diabetes. DATA SOURCES: A retrospective chart review of a non-random sample of 27 patients with amputation and 110 randomly selected patients without amputation during the same time period. Chi-square, t-tests and logistic regression were used to analyze the data. CONCLUSIONS: There was no significant difference between the group with amputation and those without amputation in age or gender. However, there was a significant difference between the two groups in the risk stratification (p = .005) and glycosylated hemoglobin levels (p = .058). IMPLICATIONS FOR PRACTICE: Nurse practitioners can effectively screen for amputation risk factors and closely monitor glycosylated hemoglobin levels of diabetic patients to reduce the incidence of vascular complications and related lower extremity amputation. PMID- 11930476 TI - [Survival of single surface ART-restorations in Zimbabwe after 3 years]. AB - A 3 year longitudinal study to assess the survival of single-surface ART restorations, was carried out among first year secondary school students in and around Harare, Zimbabwe, during the period 1993-1996 and 1994-1997. The treatments were provided by dentists and dental therapists. The evaluation was carried out by independent evaluators, one of which was an expert from the USA. Actuarial life-table analysis was applied for survival estimations. The survival of ART restorations after 3 years was 85.3% in 1993-1996 and 88.3% in 1994-1997. The 3-years survival percentage per operator ranged from 96.1% (1993-1996) and 94.3% (1994-1997) for one of the dentists to 69.3% (1993-1996) and 65.4% (1994 1997) for one of the junior dental therapists. In 1994-1997, reasons for failure were predominantly operator and material related. Secondary caries was observed in 0.5% of the ART restorations evaluated. The 3-years survival outcomes of single-surface ART restorations are promising. The study has demonstrated that ART with glass-ionomer restorative material provided quality restorative dental care to these student populations. ART has become one of the treatment modalities available to oral health workers in managing dental caries. There is a need for further research into the efficacy of the ART approach and into its application in various types of communities. PMID- 11930477 TI - [Latex glove allergy in dental practice]. AB - Skin reactions due to the use of latex examination gloves occur frequently in dental practice. They consist of immediate type I allergy to natural latex protein, type IV delayed hypersensitivity to rubber additives and irritant dermatitis. Type I allergy to latex protein can cause contact urticaria, angioedema, rhinoconjunctivitis and asthma in the dentist, the dental assistant and also in the patient. Exceptionally, latex allergy can cause a life threatening anaphylactic reaction. Cross reactions to tropical fruit can occur in the latex allergic patient. Type IV allergy causes allergic contact dermatitis. Rubber additives are also present in synthetic rubber. Glove advice for latex allergic patients are presented. PMID- 11930478 TI - [Children with ADHD]. AB - An ever increasing number of hyperactive and impulsive children receive the diagnosis Attention-Deficit Hyperactivity Disorder (ADHD). The relatively high prevalence, negative prognosis, and assumed inheritable nature of this behavioral disorder make ADHD one of the most prominent child diagnoses to date. This article discusses the behavioral characteristics, etiology, diagnostics and treatment of ADHD in school-age children. Finally, some suggestions for managing ADHD in the dental practice are given. PMID- 11930479 TI - [Syndromes 11. Treacher collins syndrome]. AB - Treacher Collins syndrome is seen once in 10.000 births. Inheritance is autosomal dominant with variable expressivity. The most prominent symptoms are antimongoloid slant of the eyelids, hypo- or even aplasia of the zygomata, very hypoplastic mandible with receding chin, deformed ear lobes and conductive hearing loss. With two to three operations a considerable improvement can be achieved. The correction of the eyelids is often the most difficult problem. PMID- 11930480 TI - [Sarcoidosis of the gingiva]. PMID- 11930481 TI - [Multiple sclerosis 1]. PMID- 11930482 TI - Home-based DM nurses: a cost-effective personal touch. PMID- 11930483 TI - Employers learn how to capitalize on DM services. AB - Employer-driven DM is on the increase. Why? Because self-funded corporations are beginning to realize the cost advantages of prevention--in terms of employee productivity as well as decreased utilization. Also, some employers want to ensure that certain key wellness or DM services will continue regardless of any changes in health coverage. While corporate interest in DM is swelling, organizations are delving into this arena in differing ways, some opting to create their own programs while others contract directly with DM vendors. PMID- 11930484 TI - Computer model creates a 'virtual world' for forecasting costs, outcomes. AB - New model creates a 'virtual world' for forecasting costs and outcomes. While predictive modeling programs are garnering most of the attention these days in the health care forecasting arena, pioneers at Kaiser Permanente have created quite a buzz with a new computer model which may be able to promise even richer information that can be tailored to specific populations. See how the model, dubbed "Archimedes," is already beginning to tackle many of the complex questions regarding chronic disease management. PMID- 11930485 TI - Study suggests video visits have significant cost, quality benefits. AB - Interactive video offers new alternatives in caring for the chronically ill. It may not be a new idea, but caring for chronically ill patients via two-way video is gaining favor as more studies surface suggesting good outcomes as well as cost advantages. In fact, our diabetes project shows that merely adding video visits to a routine schedule of in-person visits is cost-efficient. And that's just for starters. See what kinds of savings are projected when video visits are actually substituted for in-person visits. PMID- 11930486 TI - New JCAHO staffing standards: you may be surprised at what you need to do. AB - As of July 1, the Joint Commission on Accreditation of Healthcare Organizations will require you to assess staffing by using specific indicators based on clinical/service and human resource outcomes. Your ED must have sufficient numbers of qualified staff to care for patients, including direct and indirect caregivers. Nurses must have the required skills to care the for patients they are assigned to. If data reveal inadequate staffing, you must take appropriate action to correct this. PMID- 11930487 TI - Here's how to make point-of-care testing a success. AB - Point-of-care testing can ensure that patients are discharged from the ED appropriately. Tests done in the ED can save up to 50 minutes in lab turnaround time. If a single individual handles the testing and results, there is less chance of error. Joint Commission surveyors will be looking for the ability to maintain quality control and proficiency testing records. PMID- 11930488 TI - Consider pros and cons of point-of-care testing. PMID- 11930489 TI - Ask staff to help choose the right nurse manager. AB - If staff are involved in selecting a new ED nurse manager, it improves satisfaction and morale. Applicants can be interviewed by a panel of staff members. Invite staff to give input on the questions you ask candidates. Allow staff to choose the top candidates, but have administration make the final selection. PMID- 11930490 TI - Reducing practice to its essentials. PMID- 11930491 TI - Coding discrepancies. PMID- 11930492 TI - Learning from those who've left family practice. PMID- 11930493 TI - Time to re-examine physician training. PMID- 11930494 TI - 11 tips for more productive billing. PMID- 11930495 TI - Going solo: one doc, one room, one year later. PMID- 11930496 TI - How to recruit new residency graduates. PMID- 11930497 TI - Reducing waits and delays in the referral process. PMID- 11930498 TI - What every physician should know about generic drugs. PMID- 11930499 TI - Time management tips that work. PMID- 11930500 TI - Employer sponsored health benefit plans under ERISA after Pegram v. Herdrich: the fiduciary duty argument and mixed eligibility versus treatment decisions. PMID- 11930501 TI - Wrongful birth and wrongful conception: a parent's need for a cause of action. PMID- 11930502 TI - Genetic research: are more limitations needed in the field? PMID- 11930503 TI - More Hippocrates, less hypocrisy: "early offers" as a means of implementing the Institute of Medicine's recommendations on malpractice law. PMID- 11930504 TI - The punishment of "health care fraud". PMID- 11930505 TI - Biomedical and behavioral research on juvenile inmates: uninformed choices and coerced participation. PMID- 11930506 TI - Banner gears up to take on S.D. attorney general. Health system's suit argues that no law requires sale proceeds to remain in state; Barnett disagrees. PMID- 11930507 TI - Up, up and away. Spending is up, but growth slows. PMID- 11930508 TI - Now I know how it feels. A retreat provides a new perspective on women in leadership roles. PMID- 11930509 TI - Makeover at Aetna lags. A leaner Aetna focuses on profitability as it slips to No. 2 among health insurers. But aggressive downsizing has yet to buoy its sagging bottom line. PMID- 11930510 TI - The family business. Illinois hospital administrator, ACHE's young executive of the year, follows in her dad's footsteps. PMID- 11930511 TI - Crime and punishment. Instead of a slap on the wrist, crooked healthcare executives now are being sentenced to prison thanks to a crackdown on white collar crime. AB - Lawmakers wondering how to handle white-collar crime might look to the healthcare industry, where crooked executives are being held criminally accountable for shady business dealings and corrupt practices that previously may have resulted in quiet dismissals. A confluence of factors has driven the crackdown, from escalating whistleblower lawsuits to a decade-long effort to halt Medicare and Medicaid fraud. PMID- 11930512 TI - Bioterrorism: introduction and major agents. PMID- 11930513 TI - AACE hyperandrogenism guidelines. AB - The problems associated with hyperandrogenism are not uncommon in primary care settings. In fact, polycystic ovary syndrome, a common cause of androgen excess, exists in 5% to 10% of women during their reproductive years, with onset typically in adolescence. The subject of this column is the AACE's Hyperandrogenism Guidelines, which review the evaluation and treatment of the disorders causing androgenic excess. PMID- 11930514 TI - Management of osteoarthritis. AB - PURPOSE: To review the classification, epidemiology, pathophysiology, differential diagnosis, and medical management of osteoarthritis. DATA SOURCES: Selected studies, review articles, rheumatology primers, and clinical practice guidelines. CONCLUSIONS: Osteoarthritis is the most common form of arthritis and the second most common cause of long-term disability among adults in the United States. It is a heterogeneous condition causing pathogenic changes that are presumably irreversible; it should not be considered part of "normal aging." IMPLICATIONS FOR PRACTICE: Medical management of OA includes various treatment modalities and should be based on the distribution and severity of joint involvement, as well as the presence of comorbid conditions. Treatment goals include decreasing stress on involved joints, limiting physical disability, maintaining or improving function, reducing pain, and avoiding drug toxicity. PMID- 11930515 TI - Esophageal motor disorders: achalasia and esophageal spasm. AB - PURPOSE: To define the esophageal motor disorders of achalasia and esophageal spasms and describe their presentation in the clinical setting. DATA SOURCES: Selected research-based articles, textbooks, and expert opinion. A case study is presented. CONCLUSIONS: The presentation of esophageal motor disorders may not be clear, particularly when the presenting symptom is chest pain. Determining whether the pain is cardiac or digestive in origin is crucial. IMPLICATIONS FOR PRACTICE: Progressive dysphagia for both solids and liquids is the major symptom of achalasia; other symptoms include regurgitation, chest pain, and nocturnal cough. Diffuse esophageal spasm typically causes substernal chest pain with nonprogressive dysphagia and odynophagia for both liquids and solids. Dysphagia related to esophageal motility is characterized by a sensation of swallowed food "sticking" in the throat or chest; there is no problem initiating the act of swallowing. PMID- 11930516 TI - Beyond sexual assessment: lessons learned from couples post radical prostatectomy. AB - PURPOSE: To share selected experiences of advanced practice nurses (APNs) who implemented a home-based nursing protocol related to psychosexual function for couples following radical surgery for prostate cancer. DATA SOURCES: Selected research-based articles, the PLISSIT Model for sexual rehabilitation counseling, and the authors' experiences. CONCLUSIONS: Five lessons related to communicating about sexuality and intimacy were synthesized from the experience, including examining knowledge and self-awareness regularly, using a structured interview guide to facilitate the process, developing a trusting relationship with the couple, attending to verbal and nonverbal cues, and providing information about the full range of sexual expression. IMPLICATIONS FOR PRACTICE: Include an assessment of sexual health as an integral part of a general health assessment. Patients do not generally volunteer information about their sexual concerns unless the subject is introduced by the APN. PMID- 11930517 TI - Partners for NP education: the 1999 AANP preceptor and faculty survey. AB - PURPOSE: To increase understanding of national trends in nurse practitioner (NP) clinical education, the Education Committee of the American Academy of Nurse Practitioners (AANP) conducted a survey at the 1999 National AANP Conference in Atlanta. DATA SOURCES: A convenience sample of preceptors (n = 87) and faculty (n = 42) out of the total attendance of 1,744 responded to a written questionnaire. CONCLUSIONS: Preceptor respondents provided data concerning the number of NP students supervised, influence of student supervision on productivity, and availability of incentives for precepting. Faculty reported placement and supervision issues, the extent of precepting in their clinical practice site, and recognition and support for this role. Faculty and preceptors disagreed about the types and number of incentives offered for accepting students as well as the congruence of clinical teaching activities and national teaching guidelines. Precepting did not appear to strongly influence preceptor productivity. External funding did not influence opportunities for clinical education. IMPLICATIONS FOR PRACTICE: In 1998, the graduates of NP programs rose by 15.8% and over half of all nursing students enrolled in graduate nursing programs were seeking a NP education. This increase in students may compromise the ability of schools of nursing to insure quality clinical education of NP students by increasing faculty workload and placing greater demands on expert preceptors in the community. Clinical education is also changing in light of changes in the health care system. PMID- 11930518 TI - Assessing diagnostic accuracy and tympanocentesis skills by nurse practitioners in management of otitis media. AB - PURPOSE: To assess healthcare provider accuracy in recognizing the physical examination findings of acute otitis media (AOM) and otitis media with effusion (OME) and technical competence in performing tympanocentesis using a simulation model. DATA SOURCES: A descriptive study of 1,271 pediatricians and 206 nurse practitioners (NPs) who viewed 9 different video-recorded pneumatic otoscopic examinations of tympanic membranes (TMs) during a continuing medical education (CME) course. Diagnostic differentiation of AOM, OME and normal was ascertained. A mannequin model of a one-two year old child was used to assess technical proficiency of performing tympanocentesis on artificial tympanic membranes. CONCLUSIONS: Overall, the average correct diagnosis by all healthcare providers was 46% (range = 25% to 71%); by pediatricians it was 50% (25% to 71%) and by NPs 42% (25% to 68%). Pediatricians and NPs correctly recognized the absence of normality 86% to 99% and 68% to 99% of the time; they over-diagnosed AOM in 7% to 58% (average 27%) and 10% to 65% of examinations (average 31%), respectively. Performance of tympanocentesis was optimally performed by 73% of NPs. IMPLICATIONS FOR PRACTICE: The distinction between AOM and OME is important for antibiotic management decisions; tympanocentesis may be useful in diagnosis of AOM in selected patients. Healthcare providers may misdiagnose AOM and OME with some frequency. Nurse practitioners and pediatricians have the skills to be trained in the tympanocentesis procedure. Interactive CME courses with simulation technology may enhance skills and lead to a willingness to change and improve diagnostic accuracy and treatment paradigms. PMID- 11930519 TI - Substance abuse by pregnant women: legal and ethical concerns. AB - Court-ordered detention and prosecution of a pregnant woman for substance abuse is an unusual but increasing occurrence in the United States. At least 24 states have attempted to prosecute a pregnant woman on a variety of charges for suspected harm to the fetus. This article reviews the issues involved including personal autonomy duty of caregivers to women and fetuses, and ethical frameworks for discussion. PMID- 11930520 TI - Psychopharmacology in pregnancy. AB - Clinicians are confronted with challenging situations when working with women who are pregnant and have a co-existing mental illness. A risk benefit assessment is helpful when identifying possible care interventions. Psychopharmaceutical intervention is a consideration when nonpharmacological interventions are ineffective or inappropriate. Informed consent based on known and unknown risks to the mother and fetus should be obtained. Literature and case reports are contradictory and not conclusive about the risks of medications used for psychiatric illnesses. This article reviews the literature and provides clinical guidelines for antipsychotic medications, antidepressant medications, mood stabilizing medications, and antianxiety medications. PMID- 11930521 TI - Clinical management of methadone dependence during pregnancy. AB - This is a review of the literature regarding the clinical management of pregnant women maintained on methadone treatment. The prevalence of opiate use, definition of opiate dependence, common concerns regarding methadone use in pregnancy, and maternal/fetal harm are addressed. Recommendations for nurses are synthesized from the clinical literature, clinical experiences, and the empirical literature. These recommendations address: antepartum issues including treatment, dosage and pharmacological considerations, medical conditions and lab tests, intrapartum issues, postpartum concerns including breastfeeding, neonatal withdrawal, and developmental effects associated with methadone. PMID- 11930522 TI - Prominence of withdrawal symptoms in polydrug-exposed infants. AB - The problem of licit and illicit drug use during pregnancy continues to be a major perinatal health issue in this country. It is estimated that 200,000 infants are born each year to women who used illegal drugs while they are pregnant. Much information is available regarding the physiologic and neurobehavioral signs and symptoms of withdrawal expressed in infants exposed in utero to individual substances such as heroin, marijuana, methadone, alcohol, barbiturates, and PCP. However, little information is available related to the signs and symptoms of withdrawal observed in infants exposed to many combinations of these drugs prenatally. Additionally, few reports discuss the most prominent signs and symptoms of withdrawal in infants exposed to one or many drugs in utero. This article describes the results of a study that provides new information regarding the most prominent signs and symptoms of withdrawal exhibited by infants exposed to polydrugs, such as alcohol, opiates, stimulants, and sedatives, during pregnancy. PMID- 11930523 TI - Understanding drug testing in the neonate and the role of meconium analysis. AB - The use of illicit drugs during pregnancy is a problem that affects a significant number of pregnant women or women of childbearing age. For many reasons, the identification of the drug exposed mother and her infant is a necessary, albeit difficult, task. This article will discuss drug testing to detect the antenatal exposure of the newborn infant to illicit drugs and review the different laboratory methods that are used and the role of meconium analysis in neonatal drug testing. PMID- 11930524 TI - Cocaine: effects of in utero exposure on the fetus and neonate. AB - In North America, an increasing number of babies are prenatally exposed to cocaine, yet the implications of cocaine use during pregnancy are not fully understood. The effects of cocaine are exerted primarily by its influence on aminergic receptors in the central and peripheral nervous systems. Developmental, physiological, and behavioral problems in infants and children are likely outcomes of maternal cocaine abuse, but these findings are confounded by concomitant use of other drugs such as marijuana and cocaine and by factors such as time, dosage, and route of cocaine intake. Different screening options exist for cocaine and its metabolites, including sampling of neonatal urine, hair and meconium need to be considered, as do the sensitivity and the ethical implications of such testing. Clinical management of cocaine-exposed infants requires attention to several issues, including: central nervous system irritation, cardiac anomalies, apnea, and feeding difficulties, as well as infant safety and follow-up postdischarge. Early detection and intervention remain the primary objectives of caring for cocaine-exposed infants. PMID- 11930525 TI - Management of menopause. AB - Clinical practice guidelines (CPG) hold great potential for providing a summary of large volumes of clinical evidence and a related set of practical recommendations. This article is part of an ongoing series presenting individual CPGs, with a summary and brief critique. The topic addressed by the two CPGs reviewed this month is the management of perimenopause and menopause. Combined, they provide guidance on the initial assessment, ongoing assessment, and pharmacologic management of menopause, as well as a comprehensive counseling program to facilitate informed decision making by women as they experience menopause. PMID- 11930526 TI - Chagas' disease (American trypanosomiasis). PMID- 11930527 TI - Brainstem conundrum: the Chiari I malformation. AB - PURPOSE: To describe the Chairi I Malformation in relation to the anatomy of the brain and spinal cord, the common manifestations of the condition, diagnostic considerations, and management for the primary care provider. DATA SOURCES: Extensive review of the world-wide scientific literature on the condition, supplemented with actual case studies. CONCLUSIONS: The adult Chairi I Malformation is an insidious congenital brainstem anomaly that consists of caudal displacement of the cerebellar tonsils, brainstem and fourth ventricle into the upper cervical space, resulting in overcrowding of the posterior fossa. IMPLICATIONS FOR PRACTICE: Due to the vague, and often ambiguous presenting symptoms of Chiari I Malformation, many patients are misdiagnosed with conditions such as multiple sclerosis, fibromyalgia, chronic fatigue syndrome, or psychiatric disorders. Patients frequently experience symptoms months to years prior to accurate diagnosis and often incur irreversible neurologic deficits. PMID- 11930528 TI - Polycystic ovary syndrome: implications for the advanced practice nurse in primary care. AB - PURPOSE: To discuss the diagnosis and management of polycystic ovary syndrome (PCOS) by the advanced practice nurse in primary care. DATA SOURCES: Selected research and clinical articles. CONCLUSIONS: The disorder is an endocrinopathy characterized by chronic anovulation, resulting in multiple ovarian cysts. Recent research suggests a genetic etiology and a close association with obesity. Patients often present with hyperandrogenism, irregular menses, and infertility. Management is directed at the alleviation of individual symptoms. IMPLICATIONS FOR PRACTICE: If left untreated, cardiovascular disease, abnormal insulin metabolism, and ovarian and endometrial cancers may develop. PMID- 11930529 TI - Inhalational anthrax: threat, clinical presentation, and treatment. AB - PURPOSE: To provide nurse practitioners (NPs) with a basic understanding of clinical presentation, transmission, diagnosis, pharmacological treatment, and post-exposure prophylaxis of inhalational anthrax. DATA SOURCES: Selected research and clinical articles and government guidelines. CONCLUSIONS: Inhalational anthrax has an incubation period of 1 to 6 days and is very difficult to diagnose early. The chest radiograph consistently reveals a widened mediastinum and pleural effusion without infiltrates. Mortality for inhalational anthrax is high, despite aggressive treatment after onset of symptoms. Delays in diagnosis contribute to the high mortality rate. IMPLICATIONS FOR PRACTICE: The potential use of aerosolized anthrax as a biological warfare weapon has renewed interest in inhalational anthrax. Primary care providers are cornerstones in the defense against biological weapons because they may be the first to recognize and report suspicious cases. PMID- 11930530 TI - Smoking cessation interventions for pregnant women to prevent low birth weight: what does the evidence show? AB - PURPOSE: To review the epidemiology of smoking cessation and low birth weight (LBW), the use of meta-analysis and cost-benefit analysis in analyzing this problem, and the financial considerations of institutionalizing smoking cessation interventions. Recommendations for clinicians caring for smoking pregnant women and suggestions for implementing the recommended strategies for smoking cessation programs are included. DATA SOURCES: Comprehensive review of smoking and low birth weight (LBW) outcomes, cost-effectiveness, and tobacco use and dependence literature; clinical practice guidelines, and a widely cited meta-analysis on smoking cessation. CONCLUSIONS: Research has shown that smoking is a significant factor in LBW outcomes and that cessation is especially critical for pregnant women. IMPLICATIONS FOR PRACTICE: Careful evaluation of the intervention research is required before designing local interventions to ensure the most effective measures are utilized. PMID- 11930531 TI - A community needs assessment for a SANE program using Neuman's model. AB - PURPOSE: To present guidelines for a community needs assessment for a Sexual Assault Nurse Examiner (SANE) program using Neuman's Systems Model. The guidelines provide advanced practice nurses with a holistic, systematic means to conduct an assessment prior to the implementation of a SANE program and facilitate collaborative work with other disciplines and agencies. DATA SOURCES: Selected research-based articles on sexual assault, classic texts and writings on Neuman's Systems Model, and community assessment models and programs. CONCLUSIONS: Sexual assault is a problem faced by almost every community. A thorough community assessment is an important first step in establishing programs that adequately meet a community's needs. Guidelines for conducting such an assessment related to implementation of a SANE program are rare, and guidelines using a nursing model were not found in the literature. IMPLICATIONS FOR PRACTICE: One program that has been successful in meeting community needs is the SANE program. A concise and organized assessment guide can reduce the necessary time and money allocated for a community assessment and can provide a basis for evaluation and research. PMID- 11930532 TI - A military challenge to managing feminine and personal hygiene. AB - PURPOSE: To examine the personal remembrances of a group of military women concerning their experiences with hygiene issues during service in a hostile environment. DATA SOURCES: Verbatim transcripts of focused interviews from 33 women who had been deployed to combat or the field were subjected to content analysis. Questions related to the womens' experiences in cleansing the body, collecting menses waste, and protecting against genitourinary infection in a hostile environment. CONCLUSIONS: Management of personal hygiene needs was often difficult and consumed a great deal of time. Hygiene issues affected women's attitudes, practice, work, morale and coping. IMPLICATIONS FOR PRACTICE: By understanding the hygiene needs and practices of military woman changes can be made that have the potential to affect health promotion and disease prevention strategies for all women. PMID- 11930533 TI - PPS transfers still being erroneously reported as discharges. PMID- 11930534 TI - Performance measures and accountability sustain PARC (patient access and revenue cycle) improvements. PMID- 11930535 TI - [Pathological examination of breast biopsy specimens]. AB - No differences for long-term disease free survival could be found between breast conserving surgery and mastectomy. Most importantly is the fact that this therapy presents a significantly higher risk for local recurrence. The characterisation of this risk is one of the most important things to do. These findings result in a widespread change in treatment of breast cancer patients. Consequently an increase in interdisciplinary working between radiologists, surgeons and pathologists could be found. Histological examinations are necessary for diagnosis and exactly evaluation of the tumor extension. Microscopic evaluation of the resection margin is of most important interest, because there is a direct connection between local recurrence and tumor infiltration of the resection margin. We performed our investigations by the use of a standardized complete embedding method with the possibility of three-dimensional reconstruction, on a cohort of 280 patients. Additionally this method allowed the detection of all relevant findings on one hand and on the other hand an evaluation of all resection margins. Our results showed a breast conserving therapy including tumor free margins was performed in 67% of the patients. But there was a second resection necessary in 57% of the cases. An extensive tumor distribution as the detection of multifocal tumor spread was the reason for mastectomy in 33%. Our findings point out the necessarily of the histological examination in the line of the complete embedding method of the breast biopsy material in order to analyse the tumor including resection margin evaluation. PMID- 11930536 TI - [Indications for percutaneous stereotactic vacuum core biopsy of the breast]. AB - As a consequence of technical improvements and an increased number of investigations in asymptomatic patients, more and more suspicious lesions are being detected in mammography. These lesions can be evaluated using different biopsy techniques. In comparison to open biopsy, stereotactic methods require less costs and time, and reduce morbidity. The introduction of the vacuum core biopsy method allowed the excision of suspicious areas through a single needle insertion. In the current literature, however, the indications for vacuum core biopsy are being discussed controversely. This article includes an introduction of this specific technique, a presentation of the BI-RADS (Breast Imaging Reporting and Data System) categories and an overview of the literature of the indications for use of the vacuum system. The "pros and cons" of vacuum core biopsy will be discussed in comparison with the alternative biopsy methods. PMID- 11930537 TI - [Stereotactic vacuum biopsy in prone and sitting position]. AB - PURPOSE: The purpose of our study was to determine the accuracy of vacuum assisted biopsy (VB) and to evaluate the side effects of this method depending on the positioning of the patient during the intervention. METHODS: Interventions were performed with the 11-G biopsy needle either in prone position (Mammotest S, Fischer Imaging, USA) on a dedicated table for breast intervention (n = 308) or in upright position with a dedicated digital stereotactic mammography system (Mammomat 3000, Optima, Siemens, Germany). VB was indicated because of indeterminate or suspicious microcalcifications (84.1%) or because of dense lesions (15.9%). All benign results found with VB have been followed up after 6 and 12 months by mammography. All malignant or histological findings discordant to imaging features have been verified by open surgery. RESULTS: Histology after VB offered 215 (64.4%) benign lesions. In neither case a carcinoma developed during follow-up. In 90 (27%) cases malignancy was found with VB. After reexcision the diagnosis of VB was confirmed in 85.6%. In 8.9% of the cases in which VB showed DCIS, histology after open surgery revealed also invasive tumor components. Technical problems occurred in 4 cases. Major side effects developed neither in prone position nor in upright, sitting position of the patient. CONCLUSION: VB is a reliable method with less side effects for the histological evaluation of breast lesions. The accuracy and complications are not depending on the type of stereotactic device or on the patients position. The accuracy of the histological diagnosis achieved with VB can be compared to that of open surgery. The relative high costs of this method may cause a problem. But compared to the costs resulting by open diagnostic surgery this disadvantage is only a relative one. PMID- 11930538 TI - [Sonographically guided core needle biopsy of the breast: technique, accuracy and indications]. AB - PURPOSE: The purpose of this retrospective analysis was to assess the diagnostic accuracy and complication rate of sonographically guided core needle biopsy in palpable breast masses, mammographically detected nonpalpable lesions, and sonographically detected clinically and mammographically occult lesions. PATIENTS AND METHODS: Sonographically guided core needle biopsy was performed in 590 lesions in 572 patients, by using an automated biopsy gun with a 14-gauge large core needle and a coaxial system. Core needle biopsy results were compared with surgical biopsy in 265 cases. 325 lesions with benign histologic diagnoses were followed up for at least 18 months. RESULTS: 234 carcinomas and 356 benign abnormalities were found in the 572 patients. Core needle biopsy reached a sensitivity of 98.7% at a specificity of 99.7%. Underestimation rates for lesions initially diagnosed as DCIS and for lesions initially diagnosed as ADH were 3/10 and 6/14, respectively. Of three false-negative results, two were immediately recognized, and one was identified at follow-up. Serious bleeding occurred in one patient (0.2% complication rate). CONCLUSIONS: This report confirms that sonographically guided large core needle biopsy is a safe, reliable and cost effective method for the assessment of both palpable and nonpalpable, mammographically and sonographically detected breast abnormalities. PMID- 11930539 TI - [MR-guided preoperative localization and percutaneous core biopsy of suspected breast lesions. Possibilities and experience on the vertically open 0.5-T system]. AB - PURPOSE: To evaluate the feasibility of performing breast interventions in a vertically open 0.5 T MR system (SIGNA SP/i, GE Medical Systems). To develop fitted equipment and to establish preoperative wire localization and percutaneous breast core biopsy as clinical routine procedures. PATIENTS AND METHODS: Initially, we applied a localization method with the patient placed in a sitting position in 31 cases using a single loop coil and a self-developed fixation device. Subsequently, 46 wire localizations and 28 percutaneous core biopsies were carried out in prone patient position using an open breast coil with an integrated biopsy device. The used instruments were either MR-compatible (18 G biopsy needle and localization wire, 14 G coaxial needle, prototype of a 16 G double-shoot gun) or MR-safe (double-shoot gun with 16 G needle). RESULTS: After biopsy we found the needle tip (18 G for a wire localization and 14 G for a percutaneous core biopsy, respectively) placed either within or close to the lesions (< 10 mm distance) for all patients. Out of a total of 66 benign lesions and 39 malignant tumors we missed the lesion (12 mm mean diameter, 4-25 mm range) during open biopsy in two cases and obtained a false negative result for one percutaneous biopsy of a 5 mm lesion. CONCLUSION: Preoperative wire localization and percutaneous core biopsy of suspicious breast lesions demonstrated by MRI can be carried out a vertically open 0.5 T MR scanner. The degree of accuracy is comparable with that of X-ray or ultrasound-guided procedures. A follow-up has to be performed in cases with a negative biopsy. PMID- 11930540 TI - [Pulmonary aspergillosis as initial manifestation of septic granulomatosis (chronic granulomatous disease, CGD) in a premature monozygotic female twin and FDG-PET diagnosis of spread of the disease]. AB - PURPOSE: To present the unusual case of chronic granulomatous disease (CGD) manifestating as pulmonal aspergillosis in female enzygotic twins during the neonatal period. To demonstrate and to discuss the complex diagnosis and the diagnostic value of FDG-PET in monitoring the spread and activity of the disease. PATIENTS: Plain chest x-ray and CT of the lung showed multiple, mostly round lesions in one of the former preterm twins at the age of 8 weeks. The girl with known ASD II was then suffering from dyspnoea and recurrent pulmonary infections. Because of the imaging findings invasive pulmonary aspergillosis due to an underlying immunodeficiency was suspected. RESULTS: Aspergillosis was confirmed histologically and microbiologically by specimens taken from an open lung biopsy. Because of coexisting granulocytic dysfunction the diagnosis of CGD was made. This diagnosis was equally confirmed by noninvasive methods in the asymptomatic sister. FDG-PET did not show any evidence of extra-pulmonary spread of disease. CONCLUSION: CGD can present as isolated pulmonal aspergillosis even in the neonatal period when an immunodeficiency is discussed. In enzygotic twins screening of the asymptomatic twin is mandatory. FDG-PET is a useful tool in screening for spread of the disease and in evaluating disease activity. PMID- 11930541 TI - [Palpable resistance in the right renal area. Xanthogranulomatous pyelonephritis with fistulization in the colon ascendens and M. psoas major]. PMID- 11930542 TI - [Restriction of high-frequency exposure of patients in MR examinations]. PMID- 11930544 TI - [From veterinarian to quality control manager. What is going to change in the Netherlands for the practicing large animal veterinarian under influence of the White Paper about food safety?]. AB - The work of farm animal practitioners will change in the coming years as a result of the White Paper on food safety. Both government and the Royal Veterinary Association of the Netherlands are working on an accreditation system for veterinarians. The veterinary practitioner is a link in the chain to achieve safe products. Where in the past emphasis was on the individual animal, it will now be on the herd or flock. The veterinarian will officially determine the health status of the farm, which in turn will play a role in the inspection procedures at the slaughterhouse. This form of farm management will become compulsory for all stockholders within the framework of the Veterinary Network for Supervision. In turn, these developments will affect the veterinary medicine curriculum. New subjects such as quality management will become increasingly important. PMID- 11930545 TI - [Human embryonic stem cells: possibilities for future cell transplantation therapy]. AB - Human embryonic stem cells are of great importance, and Parkinson's disease is given as an example of a condition that could benefit from the development of stem cell-based transplantation therapies. The reason for this is fairly obvious: the disease is caused by the loss of only one cell type from the brain that has one major function, namely the production of dopamine. Replacement of these cells should in principle cure the disease. But what are stem cells and how far is scientific research from being able to offer stem cell-based therapy in the clinic to patients suffering from Parkinson's disease, and other chronic diseases? These questions are addressed here together with a critical evaluation of short and long-term clinical perspectives, and a discussion of possible alternatives such as adult stem cells. PMID- 11930546 TI - [Proposal of the European Commission concerning documentation of all veterinary drugs does not contribute to food safety]. PMID- 11930543 TI - [Minimally invasive surgical procedures of the breast: comparison of different biopsy systems in a breast parenchymal model]. AB - PURPOSE: The purpose of our study was to evaluate various systems for large core biopsy. First to determine the volume of specimen that can be removed with each minimal-invasive method and second to compare their consistency. With respect to the optimum of volume which can be acquired the 12-G system should be optimized by special modifications. METHODS: Examinations were performed on a phantom. We used 8 automatic, one sequential-automatic and 4 halfautomatic highspeed core biopsy systems of 16-, 14- and 12 G as well as the 11G vacuum biopsy. The weight and consistency of the specimen was determined in every case. The 12G system was optimized by enlargement of the biopsy chamber. RESULTS: The 11G vacuum biopsy exhibited best results concerning the weight of the specimen (93.7 +/- 13.5 mg). The results achieved with high speed core biopsy varied with the needle diameter as well as with the technique. Semi- and halfautomatic systems were superior to automatic systems. Best results among highspeed core biopsy systems were achieved with the modified 12G biopsy needle (41.3 +/- 4.4 mg). CONCLUSION: As expected, largest volume of specimen can be removed with vacuum biopsy. High speed core biopsy systems are inferior to vacuum biopsy. Nevertheless, the amount of volume achieved can be increased to 22.4 mg by using 14G needles and to 41.3 mg by using 12G needles. The modified 12-G needle may be considered as an less expensive alternative to 11G vacuum biopsy. PMID- 11930547 TI - [DLV Advice Group starts together with Wageningen UR project Quality and Safety of Animal Feed]. PMID- 11930548 TI - [Stable dust, lung health and PRRS diagnostic]. PMID- 11930549 TI - [Function cellular adhesion molecules determine migration pattern of neutrophil granulocytes in the lung]. PMID- 11930550 TI - [Symposium "Animal health: question of European legislation?" "Human, animal, nature and economy should be in balance"]. PMID- 11930551 TI - [Veterinary care delivery]. PMID- 11930552 TI - [Animals in intensive farming should receive rights]. PMID- 11930553 TI - [Clinical research of the cardiovascular system]. PMID- 11930554 TI - ['The panting farmer with a cow foaming at the mouth and a diagnosis out of the old box?']. PMID- 11930555 TI - [Leave him lying for a night...]. PMID- 11930556 TI - [Conclusion of notice and new law about 'independence']. PMID- 11930557 TI - [Veterinarians e-mail....]. PMID- 11930558 TI - [Procalcitonin and PaO2/FiO2 ratio as predictors of mortality in the early postoperative period after esophagectomy]. AB - OBJECTIVE: Predicting outcome in critical care remains difficult. One factor making the task difficult is, that the time elapsed between the onset of symptoms and admission to the intensive care unit is often unknown. The aim of this study is to evaluate the early course and predictive value of organ dysfunction monitored by Multiple Organ Dysfunction Score (MODS) and serum procalcitonin (PCT), microalbuminuria (M:Cr) following oesophageal tumor resection. MATERIALS AND METHODS: In our prospective study, 79 patients (67 survivors, 12 non survivors) are involved all had elective oesophageal tumour resection. MODS was monitored daily (t1, t2, t3). Serum PCT levels were determined 24 hourly (t0, t24, t48, t72). M:Cr was measured before (tp), and after surgery (t0, t6, t24, t48, t72). For statistical analysis Wilcoxon rank sum test, Mann-Whitney U test, receiver operating characteristic curve analysis and logistic regression analysis were used. RESULTS: Significantly higher MODS were observed in patients who died. As of organ dysfunction, PaO2/FiO2 ratio was below normal and remained significantly lower in non-survivors at t1, t2, t3. Serum PCT at t0 was normal in both groups. Levels at t24 increased significantly, and values were significantly higher in non-survivors. Levels remained elevated at t48 and t72 without statistically significant difference between the two groups. Preoperative M:Cr was normal. At t0 levels increased significantly, but then at t6-72 levels returned to normal without significant differences between survivors and non survivors. CONCLUSIONS: The PaO2/FiO2 ratio gave clear differentiation between survivors and non-survivors. It seems to be a sensitive measurement for predicting outcome following oesophagectomy. Routine measurement of serum PCT in the postoperative period may help predicting outcome but further studies are required. PMID- 11930559 TI - [Surgical aspects of histologically unproven space-occupying lesions of the pancreas head]. AB - A suspected, but unproved malignant tumor in the head of the pancreas is a fairly common problem for surgeon. Even intraoperatively, differentiation between chronic pancreatitis and carcinoma is difficult. We try to give guidelines about what can be done with a pancreas head mass intraoperatively without previous cytology or histology. When do we have to achieve definite diagnosis at all costs, and how can we achieve it? Results of 40 intraoperative aspiration cytologies, performed for suspected pancreatic cancer were analysed. All of them were controlled by histology in the resected specimen. Intraoperative biopsy was false negative in 12.5% and the diagnosis was uncertain in 35%. These data show that a benign result by itself never excludes the presence of malignancy. If pathology will alter our decision about resection, all efforts should be made to confirm the diagnosis. On the other hand, in a case of a mass lesion with obstructive symptoms, cytology does not alter the need for surgical decompression, and some kind of resection is a reasonable option, even in case of chronic pancreatitis. Nihilistic approach in the case of pancreatic head mass with suspected but unproved malignancy is not justified. Pancreatoduodenectomy should be performed for all tumors even without histologic confirmation if an experienced team can perform it with low postoperative morbidity and mortality. PMID- 11930560 TI - [Fournier's gangrene: early diagnosis is the best remedy]. AB - Despite the broadening knowledge about Fournier's gangrene, its mortality rate is still considered very high. The authors describe the etiology, pathomechanism, the possible pathways of progression, and the causes and predisposing factors of the disease. Besides the traditional--surgical and antibiotic--therapy, complementary ways of treatment are described. Between 1994 and 2000, ten patients were treated for Fournier's gangrene in our department. We present our experience, the early signs and the way clinical symptoms develop. It is important to observe the early severe general condition and initially minimal local changes for early diagnosis in improved survival. PMID- 11930561 TI - [Continuous autotransfusion during liver transplantation]. AB - The advantages and disadvantages of continuous autotransfusion during liver transplantation are investigated in our study compared with blood saving and traditional cell saving techniques. Patients were divided into three groups in this retrospective study; Group 1 (n = 14): continuous autotransfusion was applied; in Group 2 (n = 14): no blood saving technique used; in Group 3 (n = 14): Haemonetics cell saver was used. In Group 1 the number of Child B patients was significantly higher than Child C patients (p < 0.05). The initial values of haemoglobin were significantly lower in Groups 1 and 3 (89 +/- 19 vs. 103 +/- 17 vs. 90 +/- 16.8 g/l; p < 0.03). During hepatectomy in Group 1 lower haemoglobin values were detected than in the other two groups (96 +/- 7 vs. 104 +/- 16 vs. 106 +/- 16.6 g/l; p < 0.05). The quantity of total blood utilisation (donor + autotransfusion) was significantly higher in Group 3 than Group 2 and in Group 1 than Group 2 (21.06 +/- 11.2 vs. 11.07 +/- 3.8 vs. 30.71 +/- 18 U; p < 0.001). Comparing the values of ACT in each group during operation periods no significant difference was found. Treatment time on the ICU of the patients in Group 3 was significantly longer than in the other two groups (11.08 +/- 7.8 vs. 9.17 +/- 3.5 vs. 26.62 +/- 14.6 days; p < 0.03). We found that applying CATS is advantageous during liver transplantation, as the device reduces donor blood requirement. No significant complication was observed. PMID- 11930562 TI - [Chronic pancreatitis combined with obliterative arterial disease]. AB - We performed a prospective study at the 1st Surgical Department, Semmelweis University Budapest between 1996 and 2000, in which we examined the common occurrence of chronic pancreatitis and obstructive vascular disease in patients admitted to our department because of either of these illnesses. There were 37 patients suffering from both diseases, all are included in our trial. Control groups were set up of 50 consecutive chronic pancreatitis patients and 50 consecutive vascular patients. In all patients with two disease chronic pancreatitis developed earlier than vascular disease. Occurrence of vascular disease is twice as common among patients with chronic pancreatitis as in normal population. Risk factors like smoking, coffee-abuse and alcohol, which might trigger both diseases, didn't vary in the three groups. This combination of diseases usually starts in the young, and causes severe and progressive deterioration in the quality of life. In four patients after operations performed because of chronic pancreatitis, peripheral arterial operation was also necessary. Operative mortality rate in the chronic pancreatitis group was 2%, in vascular patients 4% and in the group with both diseases it was 16%. We found in our study that there is relation between chronic pancreatitis and peripheral obstructive vascular disease. Further examinations are necessary to examine the etiology. In patients with chronic pancreatitis we must always think of the possibility of later obstructive vascular disease. PMID- 11930563 TI - [Femoral neuropathy caused by retroperitoneal hematoma]. AB - Iliacus hematoma causing femoral neuropathy are mostly seen as a result of anticoagulation. Hematoma can occur even while correctly managed anticoagulant treatment because of drug interactions and/or coagulopathies caused by liver disease. Between 1999 and 2001 a 81 and a 68-year-old man were treated in our department because of retroperitoneal hematoma on the left side. Spinal disease and discus abnormalities were excluded with abdominal ultrasound, CT and MRI. The hematomas were operated on because of the progressive complaints and in one patient because of fever and sepsis. After the operation the patients were fast mobilised with help. To decrease the damage and to be cost-effective, hematomas causing neuropathy or other complications should be drained or operated on as soon as possible. PMID- 11930564 TI - [Laparoscopic transcystic biliary calculus extraction with Dormia basket]. AB - We describe transcystic common bile duct (CBD) stone extraction using Dormia basket in patients with preoperatively unexpected CBD stones during laparoscopic cholecystectomy (LC). We perform intraoperative cholangiography (IOC) selectively. Stone extraction was successful five times and we converted to open procedure in one patient because of the size of the stone. Mild postoperative acute pancreatitis developed once. No other complication was detected. Procedures performed if CBD stone is detected intraoperatively are evaluated in the article, including indications and difficulties of the Dormia basket method. We suggest the introduction of the method as an alternative treatment for unexpected CBD stones during laparoscopic cholecystectomy. PMID- 11930565 TI - [First experience with transjugular intrahepatic porto-systemic shunt (TIPS)]. AB - Transjugular intrahepatic portosystemic shunt (TIPS) is a radiological interventional procedure for the treatment of portal hypertension. This is a percutaneous method that lowers the elevated portal pressure by creating an intrahepatic shunt between the portal vein and one of the hepatic veins. TIPS carries lower risk than surgical portosystemic shunt, and prevents rebleeding from oesophageal varices more effectively than sclerotherapy. Since September 1999 the authors performed--with revisions--30 TIPS procedures in 20 patients. (Some patients needed more than one attempt.) We summarise our practice in patient selection, preoperative evaluation, followed by a detailed description of the technique of TIPS. Intensive monitoring and special issues of anaesthesia are also described. Technical success rate was 95%. Ninety-three percent of patients improved clinically. Despite some complications, no immediate mortality was noted. Two patients died within 30 days (both Child C). Average portosystemic gradient was reduced from 19.4 Hgmm to 9.6 Hgmm. No esophageal varix rebleeding occurred with patent shunt, and no ascites drainage was needed, either. 17% of patients developed encephalopathy requiring treatment. Our short follow-up time (average 6 months, longest 21 months) does not allow correct comparison with the 2-years survival of severe cirrhotic patients, but according to the literature TIPS does not change it significantly. PMID- 11930566 TI - The health of women: a call to move APN. Practice out of the exam room and into the community. PMID- 11930567 TI - Infectious diseases: echinococcosis (hydatid disease). PMID- 11930568 TI - Chronic obstructive pulmonary disease. AB - The GOLD Report provides valuable guidance for persons who treat persons with COPD, whether they are treating the actual disease or other comorbidities. Because COPD is such a prevalent disease and an important health problem, this set of guidelines should have broad application, in many settings. PMID- 11930569 TI - Prevention in the inpatient setting. AB - PURPOSE: To review the literature on and describe approaches to implementing health promotion and disease prevention practices in the inpatient setting. DATA SOURCES: Comprehensive review of nursing and medical literature, clinical practice guidelines, and U.S. government documents. CONCLUSIONS: Preventive screening has been shown to reduce morbidity and mortality from many diseases. Health promotion and disease prevention activities form the foundation of practice for many nurse practitioners (NPs). As increasing numbers of NPs begin to practice in the acute care setting, it is important they incorporate prevention into the inpatient setting. IMPLICATIONS FOR PRACTICE: Because there are many barriers that impact appropriate preventive screening, health care providers must take advantage of every patient encounter including those in the inpatient setting. Although it may not be feasible to actually perform certain screening exams while a patient is hospitalized, it is feasible in many cases to take a few minutes to review recommended preventive screening. PMID- 11930570 TI - Ovarian cancer strategies for nurse practitioners. AB - PURPOSE: To review and outline practical strategies for the effective detection and prevention of ovarian cancer. DATA SOURCES: Selected scientific literature, government consensus findings, and the author's experience. CONCLUSIONS: Ovarian cancer is the fourth most common cause of cancer death in American women, ranking behind lung, breast, and colorectal cancer. Seventy-five percent of ovarian cancers are currently diagnosed at an advanced stage. IMPLICATIONS FOR PRACTICE: No cost-effective screening methods are currently available. The battle to beat ovarian cancer is based on four strategies: identification of risk factors, appropriate screening methods, early detection, and prevention. PMID- 11930571 TI - Depression in post-acute myocardial infarction patients. AB - PURPOSE: To identify strategies for nurse practitioners to diagnose and treat depression following acute myocardial infarction (AMI) and prevent subsequent cardiac events. DATA SOURCES: Selected articles in the medical and psychiatric literature and government consensus guidelines. CONCLUSIONS: Between 8% and 45% of patients who have suffered an AMI exhibit symptoms of major depression. Depression is an independent risk factor for cardiovascular disease and causes higher morbidity and mortality rates among patients with coronary artery disease. IMPLICATIONS FOR PRACTICE: Early diagnosis and aggressive treatment are crucial to reducing cardiac related morbidity and mortality. Cardiac rehabilitation programs, psychotherapy and counseling, and pharmacotherapy are effective in the management of AMI depression. PMID- 11930572 TI - Peyronie's disease: assessment and treatment. AB - PURPOSE: To describe the assessment, diagnosis, appropriate testing, and available treatment options for Peyronie's disease. DATA SOURCES: Selected articles from the medical literature, standard textbooks and online resources. CONCLUSIONS: Clinical hallmarks of the disease include palpable, fibrotic plaques, temporary painful, penile curvature during erection and decreased rigidity of the penis. IMPLICATIONS FOR PRACTICE: Treatment options range from watchful waiting to surgery. Several promising medications, including vitamin E, are being investigated. PMID- 11930573 TI - Asthma care by nurse practitioners in the United States. AB - PURPOSE: To survey nurse practitioners (NPs) in the United States on their level of input to asthma care prior to enrolling in The National Asthma and Respiratory Training Centre (NARTC) specialist asthma education program in the UK. DATA SOURCES: Responses of 134 (66%) of the 202 NPs who self-selected for enrollment in the NARTC program on a preliminary questionnaire. CONCLUSIONS: Nearly all (133 or 99%) currently work with asthmatics and 122 (91%) prescribe asthma drugs. Only 92 (69%) measure peak flow(PF), 84 (63%) check inhaler technique, 76 (57%) teach use of a home PF meter and diary, and 63 (47%) provide written self-management plans on any regular basis. Many have not established follow-up procedures (56 or 42%), or evaluated their asthma management practices (55 or 41%). IMPLICATIONS FOR PRACTICE: Successful asthma management requires correct medication, systematic follow-up, patient education, and self-management. Specialist asthma training should encourage practitioners to combine effective drug usage with a long-term preventative approach. PMID- 11930574 TI - "How to get along with doctors and other health professionals". AB - All nurses will from time to time find themselves in negative interactions with doctors and other healthcare professionals. By using positive communication techniques the nurse can promote healthy interpersonal interactions and a positive atmosphere. By selecting responses rather than responding spontaneously difficult situations can be managed and a professional environment maintained. PMID- 11930575 TI - Re: JPN article fall 2000 "Once a nurse always a nurse". PMID- 11930577 TI - Hypothermia emergency tips. Knowing these few tips could save someone's life including your own. PMID- 11930576 TI - So your patient is Latino.... PMID- 11930578 TI - New education program on cardiovascular disease launched for Hispanic community. PMID- 11930579 TI - Treating tobacco use and dependence. AB - This month, the Clinical Practice Guideline (CPG) column reviews the recently published guideline, Treating Tobacco Use and Dependence. This set of recommendations is available in a quick reference format and a more comprehensive clinician's guide that includes detailed information on the methodology and references used to develop the guidelines. Both are available in electronic and hard copy versions through a variety of sources, including the National Guidelines Clearinghouse, the Centers for Disease Control and Prevention, and the National Cancer Institute. PMID- 11930580 TI - Brucellosis (Mediterranean fever, Gibralter fever, Malta fever, Cyprus fever, undulant fever, typhomalarial fever). PMID- 11930581 TI - Prescribing for seniors: it's a balancing act. AB - PURPOSE: To present a rational approach to prescribing for seniors, balancing the need for and number of medications. DATA SOURCES: Selected evidence-based reviews and research articles and the author's own experience. CONCLUSIONS: Whenever possible, alternatives to medication therapy should be considered as the initial treatment of choice; however, medications should be prescribed when indicated and not withheld due to a patient's age. Reducing the number of medications reduces the risk for adverse drug reactions and drug-to-drug interactions. IMPLICATIONS FOR PRACTICE: Careful review of an older patient's medication list prior to initiating new therapy is necessary. Follow-up care to review the efficacy and monitor any potential side effects is crucial. Timely discontinuation of a drug when therapeutic usefulness is surpassed is equally important. PMID- 11930582 TI - Postmenopausal bleeding: a practical approach. AB - PURPOSE: To provide nurse practitioners (NPs) with a practical approach for the management of postmenopausal bleeding in office settings. DATA SOURCES: Selected scientific literature and the author's own experience. CONCLUSIONS: Postmenopausal bleeding should be considered cancer until proven otherwise, despite the fact that abnormal pathology is found in only 15% of endometrial biopsies. IMPLICATIONS FOR PRACTICE: Post menopausal bleeding can be safely evaluated and managed by NPs using aspiration endometrial biopsies and/or transvaginal ultrasound. Hospitalization for dilation and curettage under anaesthesia can often be avoided. PMID- 11930583 TI - The role of human papillomavirus in screening for cervical cancer. AB - PURPOSE: To review the options for effectively screening for cervical cancer, including human papilloma virus (HPV) identification, cytologic screening, colposcopy, or a combination approach. Current pathophysiology, diagnostic criteria, treatment approaches, and patient preparation and education related to cervical cancer screening and prevention are also included. DATA SOURCES: Comprehensive review of current literature, including research and review articles. CONCLUSION: Because the Papanicolau (Pap) smear is a screening tool, not a diagnostic tool, further studies must be done to identify the actual nature of discovered abnormalities. Of particular concern is the classification of atypical squamous cells of undetermined significance (ASCUS), which may simply indicate inflammation, or may be the first indicator of serious pathology. Following ASCUS Pap smears with HPV screening will allow for a clarification of the best approach to treatment. A screening algorithm supported by a review of the literature is proposed. IMPLICATIONS FOR PRACTICE: Cervical cancer is a preventable disease caused by certain forms of HPV. Current screening protocols are based on the use of the Pap smear; and in areas where this test is routine and available, morbidity and mortality rates have dropped dramatically. Many women throughout the world and in underserved regions of the U. S. do not have adequate access to routine screening with Pap smear technology. As long as women continue to die needlessly of cervical cancer, more comprehensive and accessible screening methods must be explored. (Cutting the unnecessary worldwide and in the U. S.). PMID- 11930584 TI - The invisibility of advanced practice nurses in popular magazines. AB - PURPOSE: To determine the image portrayed in the media of nurses and particularly nurse practitioners (NPs) compared with other health care providers. DATA SOURCES: An analysis of advertisements and articles in popular magazines aimed at female, male, and gender-neutral audiences was conducted between 12/99 and 06/00. Relationships between target audience and content of advertisements and articles, as well as portrayal of health care providers were also explored. CONCLUSIONS: A review of 100 consecutive advertisements for health-related products and 96 consecutive health-related articles in popular magazines revealed that the media continues to overwhelmingly depict and promote physicians as the source of health care and health-related advice. IMPLICATIONS FOR PRACTICE: Because when there are few references to a group, such as NPs, it is reasonable to assume group members have little influence, standing, and authority, NPs should be concerned about the implications of these findings. Strategies for increasing the visibility of NPs as credible and valuable members of the health care team are shared. PMID- 11930585 TI - Teaching spirituality to nurse practitioner students: the importance of the interconnection of mind, body, and spirit. AB - PURPOSE: To describe the author's experience in locating and implementing a spiritual assessment tool in teaching a first semester clinical nurse practitioner (NP) course. DATA SOURCES: Selected articles and responses of NP students to the use of the tool. CONCLUSIONS: A spiritual assessment is an important component of a comprehensive health assessment. The use of a formal structured protocol provides a framework for beginning students to become comfortable with sprirituality and spiritual assessments and to begin to recognize spiritual distress in clients. IMPLICATIONS FOR PRACTICE: The protocol used in the course is very extensive and might not be practical in a busy practice setting. An alternative, 4-point assessment is provided. PMID- 11930586 TI - Nurse practitioner peer review: process and evaluation. AB - PURPOSE: To describe the initiation and evaluation of a nurse practitioner (NP) peer review program for a group of 15 NPs practicing at a Veteran's Affairs Medical Center. DATA SOURCES: Using a standardized review form, 15 NPs performed a peer review on a total of 163 medical records; each review was re-reviewed by 2 researchers who were also NPs. To determine the congruency of the reviews among the 15 NPs, the data were analyzed with Intraclass correlations (ICC) and ANOVA. CONCLUSIONS: A relatively low ICC (r = 0.37) was found for the 15 NPs. The ANOVA indicated that there were significant differences among the NPs (F = 11.92, p < .0001). IMPLICATIONS FOR PRACTICE: The outcome of a peer review process depends upon the motivation and values of the NPs, practice sites, and standardization of the format, as well as the degree of collective participation. Peer review can reveal charting deficiencies as well as identify topics for continuing education and risk management programs. PMID- 11930587 TI - Professional practice: what to do if you are sued. PMID- 11930588 TI - Clarifying authorship. PMID- 11930590 TI - The pleasure of your company: attitudes and opinions of preceptors toward nurse practitioner preceptees. AB - PURPOSE: To survey the attitudes and opinions of preceptors about their experiences with nurse practitioner students. DATA SOURCES: Original research using a mailed questionnaire designed by the researchers to 196 physician and nurse practitioner preceptors (23% response rate). CONCLUSIONS: Preceptors desired faculty support and feedback, wanted involvement in shaping the curriculum, and had strong opinions about the precepting experience. IMPLICATIONS FOR PRACTICE: The attitudes and opinions of preceptors are extremely important to the availability of a qualified and motivated pool of preceptors, the clinical growth of the students, the quality of the student's learning experience, and the survival of nurse practitioner programs. PMID- 11930589 TI - Challenges and strategies for initiating a nursing facility practice. AB - PURPOSE: To provide a summary analysis of five case reports describing the challenges and strategies of nurse practitioners' (NPs') first-year experiences on initiating an effective role in a nursing facility practice. DATA SOURCES: Original qualitative research analyzing written journals of five NPs and written notes from two loosely structured group discussions among project participants. CONCLUSIONS: Two broad themes emerged: Figuring it Out and Responding/Getting a Handle on Things. Common sense and good resources were identified as critical. IMPLICATIONS FOR PRACTICE: Strategies to promote ease of role transition are essential for NPs in the nursing facility. Further, in addition to a focus on care of frail older adults, emphasis on the culture of and strategies for nursing facility visits, ethical issues, and ethical decision making are important components of NP educational programs. PMID- 11930591 TI - Advances in the treatment of hepatitis C: combination antiviral therapy with interferon alfa-2b and ribavirin. AB - PURPOSE: To provide nurse practitioners with the information to manage patients with chronic hepatitis C (HCV) receiving a new combination drug therapy containing ribavirin and interferon alfa-2b. DATA SOURCES: Reviews of clinical trial results including large multicenter trials, Centers for Disease Control and Prevention documents, data from the drug manufacturer. CONCLUSION: This new therapy offers the potential for HCV remission or complete cure of the HCV infection. Although virologic responses are markedly improved with combination therapy, the side effects associated with combination therapy warrant regular patient monitoring, management, and medical intervention when clinically indicated. IMPLICATIONS FOR PRACTICE: Combination therapy does not significantly worsen the side effects associated with mono-therapy, which are predictable, manageable, and reversible. However, proper patient education, symptom management, vigilance for serious side effects, and monitoring of hematologic parameters are critical to patient outcome. PMID- 11930592 TI - Urinary incontinence: a classification system and treatment protocols for the primary care provider. AB - PURPOSE: To provide nurse practitioners (NPs) with an objective system for classification of urinary incontinence and protocols for treating the condition in the primary care setting. DATA SOURCE: Clinical practice guidelines and the author's clinical experience. CONCLUSIONS: By using tables of classifications of urinary incontinence, management strategies, and protocols, NPs can successfully manage mild urinary incontinence in primary care settings in a cost-effective manner. IMPLICATIONS FOR PRACTICE: A thorough history and objective classification of the severity of incontinence provides the framework for treating urinary incontinence in a methodical and effective manner. PMID- 11930593 TI - Screening patients for latex allergies. AB - PURPOSE: To review the reasons for the recent increase in latex allergies and the five possible routes of exposure to latex. DATA SOURCES: Selected review articles from print and electronic sources; author's clinical experience; case studies. CONCLUSIONS: Latex allergies are now being documented in groups not previously considered to be at risk due to increased screening in primary care settings. IMPLICATIONS FOR PRACTICE: Nurse practitioners should question about latex sensitivity, flag the charts of latex-sensitive clients, counsel those with a suspected latex sensitivity, refer those with known latex allergy to an allergist, and take the necessary precautions to move toward a latex-safe environment. PMID- 11930594 TI - Nurse practitioners: a comparison of rural-urban practice patterns and willingness to serve in underserved areas. AB - PURPOSE: To determine nurse practitioners' (NPs) practice patterns and willingness to practice in underserved areas in both rural and urban settings in a largely rural state. DATA SOURCES: A census of all NPs holding a Pennsylvania license and providing addresses in Pennsylvania or one of the contiguous states was conducted in 1996. The ZIP codes of practice sites were matched with 1990 census data. CONCLUSIONS: Nurse practitioners in rural areas are more likely than their urban counterparts to provide primary care in primary care practice settings, they see more patients per week, and they are more likely to be the principal provider of care for a higher percentage of their patients. Experience with managed care contracts is greater for urban NPs as is their willingness to practice in urban underserved areas. Rural NPs were more willing to practice in rural underserved areas than their urban counterparts. IMPLICATIONS FOR PRACTICE: Access to primary care continues to be a concern in rural areas. The increasing market penetration of managed care and the deficit of primary care providers in rural environments may lead to increased opportunities for NPs in the rural health care delivery system. PMID- 11930595 TI - Documentation of care outcomes in an academic nursing clinic: an assessment. AB - PURPOSE: To assess documentation of client data collected at an academic nursing clinic using the Wilson and Cleary Health Related Quality of Life (HRQOL) conceptual model as a framework. DATA SOURCES: A chart audit of 100 randomly selected active client records was conducted. CONCLUSIONS: Although several significant HRQOL variables were documented, data regarding general health perception and quality of life were not present. The HRQOL conceptual model provided an appropriate structure for evaluating the documentation. Further effort must be made to include key HRQOL dimensions in the clinic's documentation system. IMPLICATIONS FOR PRACTICE: Documenting the quality of care provided in nursing clinics is essential in order for other professionals and the public to recognize nursing professionals as accountable and credible. This project formed the basis for a computerized outcomes-based client record system. PMID- 11930596 TI - Ohio APNs: lead poisoning knowledge and practices. AB - PURPOSE: To determine lead poisoning knowledge and practices of Ohio's advanced practice nurses (APNs). DATA SOURCES: Original research conducted by mailed survey sent to all APNs registered in Ohio (N = 1448). The response rate was 72%. APNs from nine specialties and 70% of counties responded. CONCLUSIONS: Respondents correctly answered approximately 80% of the questions. Knowledge gaps in primary prevention strategies were noted. Of APNs serving pregnant women and/or children < 6 years, 57% screen for lead poisoning and 16% report no one provides lead education. IMPLICATIONS FOR PRACTICE: Lead poisoning is a preventable health problem. Education of APN students and preceptors is needed in the areas of primary lead poisoning prevention issues and practice protocols. Results from this study can help focus educational efforts and outreach programs. PMID- 11930597 TI - A critical review of current nursing faculty practice. AB - PURPOSE: To critically examine the current literature on nursing faculty practice, using the National Organization of Nurse Practitioner Faculties (NONPF) Guidelines for Evaluation of Faculty Practice, and to examine faculty practice models' strengths, weaknesses, and barriers. DATA SOURCES: Thirty-five articles describing models of faculty practice were identified through an exhaustive search on CINAHL and Medline. Two NONPF monographs on nursing faculty practice were used as guidelines for the critical review. CONCLUSIONS: Faculty practice has become an integral component of faculty-role expectations at many schools of nursing. Workload, especially without adequate compensation, remains a hindrance to practice. The value of faculty practice time and expertise has not been sufficiently demonstrated. Integration of practitioner, educator and researcher roles remains extremely difficult and sometimes elusive. IMPLICATIONS FOR PRACTICE: Faculty practice offers many advantages to schools of nursing, including educational and research opportunities for faculty and students, as well as practice sites and affordable community healthcare. Providing health care in the community presents an opportunity for independent and collaborative practice. To fully utilize the great research opportunities provided by faculty practice, more emphasis must be placed on gathering and analyzing descriptive data. PMID- 11930598 TI - Trauma risks and prevention strategies for snowboarders. AB - PURPOSE: To provide primary care providers with current information about the risk of injury and trauma prevention measures for snowboarders. DATA SOURCES: Selected research articles and Internet sources. CONCLUSIONS: Beginners are at increased risk for snowboarding injuries. Lower extremity injuries have been reduced in recent years by the introduction of the hybrid boot for beginners. Little has been done to reduce the significant impact of head injuries and wrist fractures to snowboarders. IMPLICATIONS FOR PRACTICE: Snowboarders comprise approximately 25%-37% of the population on ski slopes. Recommending the use of helmets and wrist guard protection to snowboarding patients is a useful trauma prevention measure. The high rate of injury to the beginner and young snowboarder makes a particularly strong argument for these safety recommendations to this patient population. PMID- 11930599 TI - Meeting national tobacco challenges: recommendations for smoking cessation groups. AB - PURPOSE: To review the literature on model smoking cessation programs and provide an overview of the practice guidelines for intensive smoking cessation groups. DATA SOURCES: Selected evidence-based literature and AHCPR/AHRQ clinical practice guidelines. CONCLUSIONS: Smoking is the leading cause of preventable death in our global society and is expected to kill more people than any other disease within 20 years. Data support that most smokers want to quit and are more likely to succeed with a combination of behavioral and pharmacological support. IMPLICATIONS FOR PRACTICE: Smoking cessation must be a priority in the delivery of patient care. Intensive interventions are more effective than brief interventions and should be available for all smokers. PMID- 11930600 TI - Ebola and Marburg hemorrhagic fevers. PMID- 11930601 TI - American College of Radiology Appropriateness Criteria. AB - (CPG) series provides an overview of one CPG each month. The overview includes a brief summary of the guideline's content, as well as the identification of some factors by which the author has critiqued it. The first article in the series reviewed the steps of CPG critique. Subsequent columns have described CPGs related to viral upper respiratory illnesses, tobacco dependence, menopause and perimenopause, and musculoskeletal evaluation. The document described in this month's clinical practice guideline column is actually a set of recommendations ranking the appropriateness of specific radiologic imaging or therapeutic options for particular conditions or presentations. The American College of Radiology Appropriateness Criteria provides a very valuable resource when ordering diagnostic imaging procedures. PMID- 11930602 TI - Providing wholistic health care for the elderly: utilization of the Shuler Nurse Practitioner Practice Model. AB - PURPOSE: To present an application of The Shuler Nurse Practitioner Practice Model with elderly patients through use of a gerontological wholistic assessment and the utilization of the model in developing a treatment plan and evaluating patient outcomes. DATA SOURCES: Selected literature from nursing and medicine, including natural, alternative, and complementary (NAC) therapies, as well as the authors' personal experiences. CONCLUSIONS: Providing health care services to the elderly involves recognition of not only physical ailments, but also an awareness of how functional status, psychological condition, social support, environmental conditions and cultural, as well as spiritual beliefs, shape the total health care needs of this vulnerable group. IMPLICATIONS FOR PRACTICE: Nurse practitioners who work with the aged could benefit from a theoretically based, wholistic assessment guide. Treatment plans that include NAC therapies and healing strategies should be considered. Outcome evaluations should reflect adherence to the elders' preferences regarding quality and end-of-life issues. PMID- 11930603 TI - Augmentation mammoplasty: implications for the primary care provider. AB - PURPOSE: To inform primary care nurse practitioners (NPs) of the risks and possible complications of augmentation mammoplasty and suggest guidelines for counseling and monitoring women who have undergone this procedure. DATA SOURCES: Selected research and clinical articles, government documents, and the author's experience. CONCLUSIONS: Because so many women have had breast augmentation mammoplasty, it is inevitable that NPs will see patients in the primary care settings who have complications related to the procedure or the type of implant. The most common complications include (a) changes in breast sensation, (b) capsular contracture, (c) calcifications, (d) mammography distortion or inaccuracies, (e) gel-bleed, (f) implant rupture or leakage, and (g) possible systemic reactions involving the immune system. IMPLICATIONS FOR PRACTICE: In addition to providing support and high quality preventive care, NPs may need to take on the task of coordinating other specialities and services when treating complications or when screening for breast cancer or implant rupture. PMID- 11930604 TI - Nurse practitioners and parent education: a partnership for health. AB - PURPOSE: To present the findings from a pilot-test of an innovative curriculum for parent education classes that included information about well child care and the care of sick children presented to substance abusing women at a residential drug treatment facility. DATA SOURCES: Selected literature and the results of a comparison of pre- and post-tests from the 37 women who received the classes over a 6 week period. CONCLUSIONS: The Healthy Children, Happy Children parent education program was successful overall in increasing the knowledge of the mothers about common skin disorders, care of infants, toddlers, and preschoolers, and basic first aid. IMPLICATIONS FOR PRACTICE: Nurse practitioners (NPs) are well suited to teach parent education classes that contain information on growth and development, well child care and diagnosis and management of acute minor conditions. The Healthy Children, Happy Children curriculum can be adapted to a variety of settings and populations. PMID- 11930605 TI - Diagnosis and management of chronic prostatitis by primary care providers. AB - PURPOSE: To describe the etiology, clinical manifestations, differential diagnosis, and treatment of chronic bacterial and chronic abacterial prostatitis (CBP and CAP respectively) in the primary care setting. DATA SOURCES: Selected research, clinical guidelines, and research-based articles in the scientific literature. CONCLUSIONS: Most cases of CBP can be appropriately diagnosed and treated in the primary care office. In the case of a diagnosis of CAP, initial therapy can be started by the nurse practitioner (NP) with referral to a urologist for refractory cases. IMPLICATIONS FOR PRACTICE: Complete eradication of pathogens in CBP is not always possible. Assisting patients to carefully follow their treatment regimens, including completion of all antibiotic therapy, will reduce the frequency and severity of symptoms. Successful management of chronic prostatitis symptoms can result in an improved quality of life and an increased ability to perform activities of daily living for patients. Chronic abacterial prostatitis may require referral to a urologist or mental health professional for co-management of symptoms. PMID- 11930606 TI - Strategies for working with elderly clients: a qualitative analysis of elderly client/nurse practitioner communication. AB - PURPOSE: To describe communication between nurse practitioners (NPs) and elderly clients. DATA SOURCES: A qualitative study using a focus group composed of 6 NPs and in-depth interviews of an additional 6 NPs who provide care to elderly clients in a variety of settings. CONCLUSIONS: The NPs described a framework of critical thinking, termed hypervigilance, which they enter before and during an interaction with an elderly client. Specific communication strategies were identified that increased the likelihood of compliance with or adherence to treatment regimens for elderly clients. IMPLICATIONS FOR PRACTICE: Hypervigilance is descriptive of actively working to merge nursing practice with traditional medical primary care practice. This finding raises the issue of the difference in practice between physicians and NPs in regards to communication with clients. PMID- 11930607 TI - [Emphasis on immune prevention of Helicobacter pylori infection]. PMID- 11930608 TI - [Mission and challenge: health care of Chinese children in the new century]. PMID- 11930609 TI - [The prevalence and mechanism of Helicobacter pylori resistance to clarithromycin in Beijing]. AB - OBJECTIVE: (1) To determine the prevalence of Helicobacter pylori strains resistant to clarithromycin in Peking. (2) To determine the genetic mutation in relation to clarithromycin resistance. METHODS: Helicobacter pylori were cultured from gastric biopsies obtained from 89 patients during upper endoscopy. Minimal inhibitory concentrations of clarithromycin were determined by Etest method. Genomic DNA was prepared by the hexadecyltrimethylammonium bromide (CTAB)-phenol extraction method. Mutations in clarithromycin-resistant strains were identified by polymerase chain reaction and restrictions analysis. RESULTS: (1) The prevalence of Helicobacter pylori strains resistant to clarithromycin was 13.5% in Beijing. (2) All twelve clarithromycin-resistant strains had the new BsaI site which is characteristic of the A2143G point mutation in 23S rRNA gene, none of the tested twenty-four susceptible strains had the A2142G or A2143G mutation. CONCLUSION: The prevalence of clarithromycin-resistant strains was not rare in Beijing. Mutations in 23S rRNA are associated with clarithromycin resistance in Helicobacter pylori. PMID- 11930610 TI - [Gene cloning and expression of outer membrane protein of Helicobacter pylori]. AB - OBJECTIVE: To construct a recombinant vector which expresses the 18 kDa outer membrane protein (OMP) from Helicobacter pylori (Hp), and exploit the possibility of obtaining Hp vaccine and diagnostic reagent kit for rapid diagnosis of Hp infection. METHODS: The gene encoding the structural 18 kDa outer membrane protein of Hp was amplified from Hp chromosomal DNA by PCR. The purified PCR product and identified plasmid pQE30 underwent restriction enzyme (Hind III, BamHI, BgLI) digestion and purified by PCR purification reagent kit, and then linked in the proportion of 4:1 (molar weight). The recombinant vector was sequenced with T7 as seqiencing primer. Homology of the determined DNA sequence was analyzed by DNA analysis software. The recombinant vector was selected and identified by restriction enzyme digestion, and then transformed into DH5 a (PREP4) Escherichia coli strain which was cultured and induced by isopropylthio beta-D-galactosideso as to determine its expressed products. RESULTS: The gene segment inserted into the recombinant vector was identified as the gene experssing the OMP of HP with a molecular mass of 18 fDa. As compared with previously reports, 2% of the gene was mutated, 1.68% of the amino acid residues was changed, and the homogeneity was about 98%. The level of soluble expression products was about 18% of total cellular protein. ELISA results showed that this objective protein could be recognized by anti-serum against Hp. CONCLUSION: The product expressed by Hp OMP gene clone has good antigenicity. The recombinant vector expressing 18 kDa OMP may be a potential source for effective protein vaccine against Hp infection and reagent kit of Hp infection diagnosis. PMID- 11930611 TI - [Effects of touch on growth and mentality development in normal infants]. AB - OBJECTIVE: To determine the effects of conducting Touch to infants upon their growth and mental development, sleeping and disease conditions, as well as the difference of effects of conducting Touch among infants of different month age. METHODS: 310 normal full-term infants aged 0-6 months were divided randomly into Touch group (n = 208) and control group (n = 102). Parents of Touch group were asked to conduct Touch to their infants 2-3 times/day, lasting 15-20 minutes every time, for at least 3 months. Intelligence tests were performed to infants of both groups at the beginning of the experiment and 6 months later (CDCC measurement table was employed). Data such as height, weight, circumferences of head and chest, sleeping, feeding and disease conditions etc. were recorded each month in a successive period of 6 months. Statistical analysis was conducted using SPSS 10.0. RESULTS: (1) The average score of mental development (MDI) was 7.4 higher, with a statistically significant difference, among the infants who began to receive touch within 3 months after birth than those infants with the same age in control group. The average score of psychomotor development index (PDI) was 5.8 higher, with a statistically significant difference, among the infants who began to receive touch within 3 months after birth than those infants with the same age in control group. Infants who began to receive touch within 3 months after birth slept remarkably better than those in control group did. No significant difference was found in scores of MDI and PDI and sleeping condition between infants who began to receive touch 3 months after birth and the infants with the same age in the control group. (2) Weight gain and growth in chest circumference in infants receiving touch within one month after birth were significantly better compared with the infants of the same age in the control group. There was no significant difference in values of height and head circumference between the two groups. There was no significant difference in values of height and head circumference between the two groups. (3) No significant difference in the incidence rates of acute respiratory infection and diarrhea disease was found between the touch group and control group. CONCLUSION: (1) Growth and mental development of infants can be improved by touch. Tthe earlier to conduct touch the better the effects. (2) Touch helps improve sleeping condition in infants. PMID- 11930612 TI - [Monitoring and intervention of infant mortality rate and fetal and infant mortality rate in Beijing, 1992-2000]. AB - OBJECTIVE: To monitor the changing trend of infant mortality rate (IMR) and fetal and infant mortality rate (FIM) in Beijing and to develop proper measures to decrease these two rates. METHODS: Dynamic series analysis was used to analyze the monitoring data of 590,833 cases of live birth, 5,835 cases of infant death, and 10,499 cases of stillbirth in Beijing during 1992-2000. The interventional measures in different phases were analyzed too. RESULTS: The IMR and FIMR in Beijing were 12.42% and 19.43% respectively in 1992 and were reduced to 6.84% and 15.79% respectively in 2000. The IMR and FIMR were declining significantly during the periods of 1992-1994, 1995-1997, and 1998-2001 (P < 0.01). The decrease of IMR by 48% was more remarkable than that of FIMR by 19%. The IMR I rural areas was higher than that in urban area. Pneumonia dropped from the position of the first death cause for infants to the fourth cause and congenital heart disease became the first cause. CONCLUSION: FIMR helps more comprehensively evaluate the effect of maternal and child health care. Monitoring and intervention of congenital heart diseases is very important. PMID- 11930613 TI - [Identification of high-pathogenicity island-harboring Escherichia coli from children with diarrhea disease]. AB - OBJECTIVE: To investigate the etiological status of high-pathogenicity island harboring Escherichia coli (HPIEC) in diarrhea diseases among children. METHODS: Escherichia coli was Isolated and identified from the stool specimens of 1,032 children with diarrhea diseases by culturing, serotyping, PCR, and colony hybridization. RESULTS: 652 strains of E. coli were isolated from the 1,032 stool specimens and confirmed by colony hybridization out of which 225 were identified as diarrheagenic E. coli, including 20 EPEC, 81 ETEC, 47 SLTEC, 74 ESIEC, and 3 EIEC strans. 112 (17.2%) irp2 virulent gene positive E. coli strains were detected from the 652 strains, out of which 24 were from the 74 ESIEC strans and 17 from the 47 SLTEC stranis. The typical clinical symptoms of diarrhea caused by HPI-haboring E. coli included anorexia (87.5%), abdominal pain (58.0%), diarrhea (75.9%, over 6 episodes, mostly of mucous stool a day, 75.9%), and fever (50.9%). CONCLUSION: High-pathogenicity island-harboring E. coli is one of the important pathogens of diarrhea diseases in children. PMID- 11930614 TI - [Study on the stroma of patients with paroxysmal nocturnal hematoglobinuria]. AB - OBJECTIVE: To investigate the relationship between the stroma of bome marrow and the abnormality of hematopoiesis in patients with paroxysmal nocturnal hematoglobinuria (PNH). METHODS: The bone marrow stromata from PNH patients was compared with those from normal volunteers in aspects of their CFU-F formation and hematopoietic sustentation for normal CD34+ cells. The stromal IL-6 and TNF alpha mRNA expression in patients and normal volunteers was also measured by RT PCR(self beta-actin as contrast). RESULTS: (1) The CFU-F formation ability in PNH patients was very similar to that in normal volunteers (P > 0.05). (2) The bone marrow stromata from patients with PNH had the same hematopoietic maintenance compared to those from normal controls (P > 0.05). (3) The stomal IL-6 and TNF alpha mRNA expression was similar in PNH patients and in normal volunteers (P > 0.05). CONCLUSION: The CFU-F formation capacity of bone marrow fibroblast was normal in PNH patients. The stromata in patients with PNH had normal hematopoietic maintainence to the normal CD34+ cells. The mRNA expression of IL-6 and TNF-alpha in PNH stromata was almost normal. PMID- 11930615 TI - [Screening for the new variant of defective receptor-binding apolipoprotein B 100]. AB - OBJECTIVE: To detect possibly existing unknown mutation(s) in nucleotide sequence coding amino acid residues 2,980 to 3,084, that may lead to familial defective apo B-100, and to provide evidence for evaluating the putative receptor binding domain of apo B-100. METHODS: The nucleotide sequence coding the amino acid resudues 2,980-3,084 of apo B gene in 341 patients with primary hypercholesteremia and of 50 controls was amplified by PCR and subjected to single strand conformational polymorphism analysis under optimized conditions. RESULTS: No abnormal electrophoretic figure was found in the samples from 341 hypercholesterlemic patients and 50 controls. CONCLUSION: (1) Point mutation causing hypercholesterlemia is unlikely to exist, or rare, if any, in the codons 2,980-3,084 of apo B-100 in Chinese. (2) Amino acid residues 2,980-3,084 may not be involved in the receptor-binding of apo B-100. PMID- 11930616 TI - [Genetic polymorphism analysis of apolipoprotein CII microsatellite DNA (TG) n (AG) m in patients with coronary heart disease]. AB - OBJECTIVE: To investigate the relationship between the polymorphism of apolipoprotein CII (apoCII) microsatellite DNA (TG) n (AG) m and coronary heart disease (CHD). METHODS: The apoCII microsatellite DNA (TG) n(AG)m genotypes of 108 patients with CHD and 231 healthy persons were detected by using polymerase chain reaction (PCR) and high voltage denatured polyacrylamid gels electrophoresis. RESULTS: The allele frequency distribution was significantly different between controls and patients with CHD (chi 2 = 19.93, P < 0.05). The allele 17 frequency in CHD group was remarkably higher than that in controls (chi 2 = 13.98, P < 0.01). The concentrations of TG and HDL-C among different genotype groups were significantly different. The levels of TG in 17/29 group and 17/30 group were higher than those in other groups (P < 0.05), while the concentration of HDL-C was lower (P < 0.05). CONCLUSION: The allele 17 of apoCII may be an independent risk factor for CHD in Han population. PMID- 11930617 TI - [Classification of intracranial dural arteriovenous fistula and its clinical signification]. AB - OBJECTIVE: To develop a practical imaging calssification of intracranial dural arteriovenous fistula (DAVF) based on imaging and its clinical significance. METHOD: The lesion positions and drainage patterns of vein in 110 patients with 121 dural arteriovenous fistulas hospitalized in Xuanwu Hospital were analyzed and a new type of classification of DAVF was developed. RESULTS: The dural arteriovenous fistulas in 110 patients were classified accrding to lesion position into DAVF of dural sinus, cavernous sinus, tentorial incisure, venous plexus at skull base, and cerebral falx; or classified according to the pattern of venous drainiage into type I (draining directly into dural vein or dural sinus), type II (directly draining into dural sinus with retrograde venous drainage into cortical vein or spinal vcein), and type III (directly draining into cortical vein or spinal vein). Then, based on a combination of these two criteria, DAVF was claassified into cavernius sinus area type I (38 cases, 69.1%) and type II (17 cases, 30.9%); dural sinus area type I (11 cases, 33.3%), type II (16 cases, 56.4%), and type III (2 cases, 10.3%); and tentorial area (20 cases), skull base vein plexus area (6 cases), and cerebral falx area (1 case), all in type III. CONCLUSION: The newly developed classification system of intracranial dural arteriovenous fistula helps analyze clinical risk and determine therapeutic procedures. PMID- 11930618 TI - [Relationship between serum 1,25-dihydroxyvitamin D and bone loss in postmenopausal women]. AB - OBJECTIVE: To study the correlations between loss of activity and/or amont of serum 1,25-dihydroxyvitamin D(1,25(OH)2D) and 25-dihydroxyvitamin D(1,25(OH)2D) and bone loss in postmenopausal women, in order to reveal the possible pathophysiology of postmenopausal osteoporosis. METHODS: The serum levels of 1,25 (OH)2D, 25 (OH) D, PTH, and E2 in 57 postmenopausal women with osteoporosis and 37 postmenopausal women without osteoporosis were measured by radioimmunology. Their Dpd/Cr in urine was measured by ELISA. The bone mineral density (BMD) at lumbar spine (L2-4) was measured in each subject by DEXA. RESULTS: The serum 1,25(OH)2D in postmenopausal oeteoporotic women and non-osteoporotic women were 18 pg/ml +/- 6 pg/ml and 31 pg/ml +/- 14 pg/ml respectively (P < 0.01) The serum 25 (OH)2D in postmenopausal oeteoporotic women and non-osteoporotic women were 32 ng/ml +/- 9 ng/ml and 46 ng/ml +/- 17 ng/ml respectively (P < 0.01). Postmenopausal osteoporotic women had a lower estrogen concentration (P < 0.01), higher serum parathyroid hormone (PTH) (P < 0.01) and higher urinary Dpd/Cr level (P < 0.01). The level of serum 1,25(OH)2D was highly correlated with BMD in L2-4 for postmenopausal women (r = 0.693, P < 0.001). There was a close relationship between the serum level of 1,25(OH)2D and serum level of 25(OH)2D, suggesting an association between the two factors. CONCLUSION: The lower bone mass density was associated with lower level of serum 1,25-dihydroxyvitamin D, accompanied by reduced serum estrogen and elevated serum PTH in postmenopausal women. Those may be among the important pathphysiological changes of postmenopausal osteoporosis. PMID- 11930619 TI - [Genetic diagnosis and prenatal diagnosis of spinal muscular atrophy by restriction endonucleases digestion of PCR product]. AB - OBJECTIVE: To establish an efficient and rapid method for genetic diagnosis and prenatal diagnosis of spinal muscular atrophy (SMA). METHODS: 11 SMA patients were detected for the deletion and mutation in the survival motor neuron (SMN) gene by restriction endonucleases digestion of PCR products based on the difference between the two homologous copies of SMN. Linkage analysis was performed using four (CA) n repeats. RESULTS: Both eon 7 and exon 8 were deleted in 10 patients. Only exon 7 deletion was found in one patient. Prenatal diagnosis of four SMA pedigrees was performed by restriction endonucleases digestion. SMN gene fragments of three fetuses were different from those of probands, and the SMN gene fragments of one fetus was the same as those of proband. CONCLUSION: Restriction endonucleases digestion of PCR products is an efficient and rapid method for diagnosis of SMA. PMID- 11930620 TI - [Dynamic change of tumor necrosis factor and endothelin during perioperative period in patients undergoing coronary artery bypass grafting]. AB - OBJECTIVE: To observe the dynamic changes of tumor necrosis factor-alpha(TNF alpha) and endothelin (ET-1) during perioperative period in patients undergoing coronary artery bypass grafting (CABG) with cardiopulmonary bypass (CPB) and to investigate their pathological significance so as to bring up feasible measure to increase therapeutic effect. METHODS: Blood samples of 32 patients undergoing CABG with CPB were drawn before operation, induction period of anaesthesia, after aortic crossclamping and declamping, by the end of surgery, and 2 hours, 8 hours, and 24 hours after surgery. The plasma TNF-alpha and ET-1 were measured by radioimmunoassay (RIA). RESULTS: (1) The plasma TNF-alpha level increased markedly after aortic crossclamping (18.3 ng/L +/- 3.4 ng/L vs 12.1 ng/L +/- 2.0 ng/L, P < 0.05) and went up to the peak value(22.4 ng/L +/- 3.6 ng/L) after aortic declamping, and then decreased. However, till 24 hours after operation, the TNF-alpha concentration maintained at a level higher than that before surgery (18.5 ng/L +/- 4.1 ng/L vs 12.1 ng/L +/- 2.0 ng/L, P < 0.05. After aortic declamping, during reperfusion of the lungs the TNF-alpha level was significantly higher than that after aortic crossclamping and before declamping (22.4 ng/L +/- 3.6 ng/L vs 18.3 ng/L +/- 3.4 ng/L P < 0.05). (2) The plasma ET-1 increased significantly after aortic crossclamping (146 ng/L +/- 20 ng/L vs 97 ng/L +/- 14 ng/L, P < 0.05) and went down after aortic declamping. Two hours after surgery, the plasma ET-1 increased again and reached the second height (134 ng/L +/- 19 ng/L), and then went down again and decreased to 92 ng/L +/- 18 ng/L 24 h after operation. CONCLUSION: (1) Attenuation of inflammatory reactions mediated by TNF alpha should be considered an important strategy for myocardial and lung protection. (2) During the perioperative period of CABG the plasma ET-1 level shows two peaks, occurring respectively after aortic crossclamping and 2 h after surgery, which may result from operation itself and ischemia-reperfusion injury. PMID- 11930621 TI - [Clinical research of primary hyperparathyroidism]. AB - OBJECTIVE: To investigate the clinical features and therapeutic procedures of primary hyperparathyroidism(PHPT) in China. METHODS: Clinical data of twenty eight consecutive in-patients with PHPT hospitalized in the Second Hospital Affiliated to Zhejiang University Medical School during the period 1968-2001 were analyzed retrospectively and compared with the domestic relevant literature. RESULTS: Twenty-eight patients underwent operative treatment, including exploratory operation of neck in 14 cases, cystectomy in one case, cancer radical operation in one case, and direct parathyroid adenomectomy in 14 cases with solitary adenoma. All cases resulted in normocalcemia after operation. The cure rate of PHPT was 100%. No early and long-term operative complications and recurrence were found. CONCLUSION: PHPT, mostly occurring among people aged 30 40, is rare in China. Osteopathy can be seen in 90% of PHPT patients. For solitary parathyroid adenoma, direct adenomectomy through small neck incision is a safe and effective alternative to conventional neck exploratory operation. PMID- 11930622 TI - [Relationship between serum CA125 level and prognosis in the patients with uterine endometrial carcinoma]. AB - OBJECTIVE: To study the relationship between serum CA125 level and prognosis of uterine endometrial carcinoma. METHODS: The serum CA125 levels in 71 patients with uterine endometrial carcinoma were measured by using microparticle enzyme immunoassay. The relationship between preoperative CA125 levels and surgical pathologic staging, histologic classification, and other prognostic factors was investigated. RESULTS: (1) The positive rate of serum CA125 in postmenopausal patients was significantly higher than in premenopausal patients. (2) The positive rate of serum CA125 was related to surgical pathologic staging. (3) Serum CA125 level was correlated not only with histological classification but also with other prognosis factors. (4) Increase of serum CA125 after operation predicted the possibility of relapse. CONCLUSION: CA125 is a useful marker in predicting the prognosis of uterine endometrial carcinoma. PMID- 11930623 TI - [Enhancement of study and administration of stem cell]. PMID- 11930624 TI - [Comments on the research of chronic obstructive pulmonary disease]. PMID- 11930626 TI - [Implantation of skeletal muscle stem cells in inhibition of fibroatrophy of ischemic myocardium: an experimental study]. AB - OBJECTIVE: To investigate the effect of stem cells derived from autogenous skeletal muscle, namely satellite cells, and implanted into ischemic myocardium on inhibition of myocardium fibroatrophy. METHODS: The left anterodecendant arteries (LAD) of 12 adult dogs were ligated so as to establish animal model of acute myocardiac infarction. Satellite cells isolated from the greatest gluteal muscle of dogs were labeled with 4' 6-dimidino-2-phenylindone (DAPI), and then infused into the ventricular myocardium of the isogenic dogs through LAD. Specimens of ischemic myocardium were taken 2, 4, and 8 weeks after myoblast implantation. Histologic sections are examined under common light microscope and fluorescent microscope. Twelve dogs were used as controls. RESULTS: In the ischemic myocardium where satellite cells had been implanted, fibroaastrophy was efeectively inhibited. The implanted satellite cells had differentiated into fully developed striated muscle cells. Vitreous degeneration and disorder of basic structure could be observed in the ischemic myocardium in the control group. CONCLUSION: The satellite cells from autologous skeletal muscle differentiate into cardiac muscle cell-like cells in the ischemic area and inhibit fibroatrophy of the ischemic myocardium after implantation, thus bringing a hope of a new cure for myocardial damage. PMID- 11930625 TI - [Allogeneic peripheral blood stem cell transplantation for high-risk leukemia]. AB - OBJECTIVE: To study the effects of allogeneic peripheral blood stem cell transplantation (allo-PBSCT) in high-risk leukemia. METHODS: From October 1995 to December 2000, 25 patients with high-risk leukemia (median age 34 years, range 5.5-52 years) were transplanted with peripheral blood stem cells from their HLA identical sibling donors. Among them, 15 patients suffered from acute leukemia (ALL) (one ph+ ALL in CR1, seven in CR2 or greater, and seven in relapse, including two in relapse after the first all-BMT), four patients suffered from chronic myelocytic leukemia (CML) (in CP2, AP, BC, and relapse after BMT respecively), 6 patients suffered from myelodys plastic syndrome (MDS), including one case of refractory anemia with excess of blasts (REAB), one case of refractory anemia with excess of blasts in trransformation (REAB-T), and 4 cases of acute leukemia secondary to MDS. The graft versus host disease (GVHD) prophylaxis included administration of cyclosporine and methotrexate. RESULTS: All patients were successfully engrafted. The median times (range) for their neutrophil returning to > or = 0.5 x 10(9)/L and for platelet returning to > or = 20 x 10(9)/L were 14 (10-18) days and 11 (7-45) days after transplantation resprctively. Grade II acute GVHD occurred in 12 patients with an incidence rate of 48%. Grade III GVHD was found in one patient (4%). No grade IV GVHD was seen. Among the 23 evaluable patients, 16 were diagnosed as chronic GVHD (70%). The actual transplant-related mortality was 16%. Leukemia relapse occurred in 6 patients, four of them received donor lymphocyte infusion (DLI) and achieved remission again. Nineteen patients were alive and disease-free with a median follow-up time of 304 (94-1,963) days. The two-year probability of overall survival, disease-free survival (DFS), and relapse rates were 64%, 58%, and 25% respectively. CONCLUSION: Allo-PBSCT decreases the relapse rate, increases the disease-free survival rate for patients with high-risk leukemia. All-PBSCT may be a better choice for patients with high-risk hematological maligmancies. PMID- 11930627 TI - [Risk factors of chronic obstructive pulmonary disease in patients with chronic bronchitis]. AB - OBJECTIVE: To explore the risk factors of chronic obstructive pulmonary disease (COPD) in patients with chronic bronchitis in following-up 8 years. METHODS: The baseline survey of the study was carried out on 1,999 patients with chronic bronchitis screened out from 67,251(15 years or more) rural people in 1992. In 2000, 1,114 patients were reexamined by simple random sampling method. RESULTS: 869 (78.0% of 1,114) with complete data entered analysis of this study. Smoking, and family history of COPD were associated significantly positively with the decrement per capita yearly of FEV1(forced expiratory volume in the first second) and/or FEV1/FVC (forced vital capacity) ratio. Ex-smoking decreased the decrement of lung function in smokers. The accumulative incidence of COPD was associated positively with smoking and family history of COPD and negatively with baseline FEV1 and FEV1/FVC. The relative risk of COPD was not significantly different between non-smokers and ex-smokers. CONCLUSION: For Chinese rural patients with chronic bronchitis, aging, smoking, family history of COPD, lower pulmonary function are independent risk factors of COPD while sex is not one. Ex-smoking can almost decrease the relative risk of COPD to the approximative risk level of non-smoking. PMID- 11930628 TI - [Activation of transcription factors and induction of cytokines from macrophages in chronic obstructive pulmonary disease]. AB - OBJECTIVE: To investigate the activation of transcription factors and induction of cytokines from alveolar macrophages in chronic obstructive pulmonary disease (COPD). METHODS: Alveolar macrophages were collected by fibrobronchoscopy from 8 patients with chronic bronchitis, 8 patients with COPD, and 8 healthy volunteers. All patients were at stable stage. The macrophages thus collected were cultured and stimulated with lipopolysaccharide (LPS, 10 micrograms/ml). The IL-8, IL-1 beta, TNF alpha and IL-6 thus produced were measured by ELISA in the supernatant. Nuclear factor-kappa B (NF kappa B), activator protein-1 (AP-1), AP-2 and AP-3 were detected by electrophoretic mobility shift assay. RESULTS: The concentration of IL-8 released from macrophages of patients with COPD at stable stage before LPS stimulation was about 3 times higher than that in the healthy control (F = 4.34, P < 0.05). The concentration of IL-8 released from macrophages in patients with COPD was increased further after LPS stimulation in comparison to that in healthy controls(F = 3.56, P < 0.05). The concentration of IL-1 beta and that of TNF alpha released from macrophages of COPD patients were further increased after LPS stimulation (P < 0.05) in the COPD patients, but there was no difference in the concentration of IL-1 beta and between the control and COPD patients before LPS stimulation. The constitutive activity of AP-1 and the activity of NF kappa B induced by LPS were higher in the patients with COPD than in the controls. CONCLUSION: The alverlar macrophages of patients with COPD at stable stage may release higher concentration of IL-8 and IL-1 beta. LPS stimulation increases the release of IL-1 beta and TNF alpha of alveolar macrophages. Enhancement of activity of NF kappa B and AP-1 may positively regulate the production of IL-8 and IL-1 beta in the airflow obstruction. PMID- 11930629 TI - [Effects of hepatitis G virus coinfection on hepatic pathological changes of patients with chronic hepatitis B: a long-term comparative study]. AB - OBJECTIVE: To observe the long-term effect of hepatitis G virus (HGV) coinfection on hepatic pathological changes of patients with chronic hepatitis B (CHB) and explore the pathogenicity of HGV. METHODS: Menghini method liver biopsy was performed on 45 patients with CHB twice with an interval of 5 years on a voluntary basis. The liver tissue of 21 cases was HGV nonstructural region 5 (HGV NS5) antigen positive by peroxidase anti-peroxidase (PAP) immunohistochemical staining assay (HGV coinfection group) in these two tests, and 24 cases were negative (HGV noncoinfection group). There was no significant difference in age, sex, course of disease, amount of serum HBV-DNA by competitive quantitative polymerase chain reaction assay and severity of hepatic pathological leisions between these two groups (P < 0.05) and they were treated with the same scheme and without sustained curative effeect. The hepatic pathological changes of the two groups after 5 years were compared and analysed retrospectively. RESULTS: There was no significant difference in basic hepatic pathological change, severity of inflammatory activity grade and fibrosis stage in these two groups. The numbers of cases of inflammatory activity grade G1, G2, G3, and G4 were 3, 7, 7, and 4 in HGV coinfection group, and 5, 8, 7, and 4 in HGV noncoinfection group in the beginning of observation; and were 3, 5, 7, and 6 in HGV coinfection group, and 4, 6, 8, and 6 in HGV noncoinfection group after 5 years. The numbers of cases in fibrosis stages S1, S2, S3, and S4 were 4, 7, 7, and 3 in HGV coinfection group, and 6, 8, 6, and 4 HGV noncoinfection group in the beginning of observation; and were 3, 6, 5, 7 in HGV coinfection group, and 4, 7, 5, and 8 in HGV noncoinfection group after 5 years (P > 0.05). CONCLUSION: HGV coinfection does not lead to the activation of hepatic pathological changes and does not speed up the progression of fibrosis in patients with CHB. HGV coinfection does not play an obvious role in long-term hepatic pathological changes of patients with CHB. HGV has at most a mild hepatic pathogenicity. PMID- 11930631 TI - [Preparation of decalcified bone matrix, bone cement and bone morphogenetic protein composite]. AB - OBJECTIVE: To investigate the most excellent composite proportion and the most convenient composite method of decalcified bone matrix (DBM), bone cement (BC) and bovine bone morphogenetic protein (bBMP) composite. METHODS: The canine DBM and the bBMP were prepared beforehand. First mix the bBMP and DBM together with the proportion of 1:25 with adsorption and directly, then add BC with different weight proportion and finally the complex material was examined by biomechanics, scan electron microscope, determination of bone inductive activity and implanted into the canine femoral defect caused by microwave-induced hyperthermia. RESULTS: The bBMP had strong bone inductive activity and had no affect for this function when combined with DBM with different method. There were irregular gaps in the composite and most of them with diameter less than 100 microns and the porosity of the composite was less than 20% when the proportion of DBM was 40%. So it was unfavorable for the new bone to form and the new blood vessels to reconstruct. The DBM and BC could not be combined together when the proportion of DBM was 80%. While the biomechanical intensity of the composite decreased and the qualities of the new bone forming and new blood vessels reconstructing improved successively when the proportion of DBM was from 50% to 75%. CONCLUSION: It is the most convenient that bBMP is combined with DBM directly while its bone inductive activity could not be affected. The composite material not only has high biomechanical intensity, but also is favorable for new bone to form and new blood vessels to reconstruct when the proportion of DBM is 50% and 60%. So it is favorable for bone repair in the places where need to bear a load. However it is also favorable for bone repair in the places where do not need to bear a load when the proportion of DBM is 75%. PMID- 11930630 TI - [Phenotypic and functional alterations of mesangial cells in patients with diabetic nephropathy]. AB - OBJECTIVE: To explore the phenotypic and functional changes of glomerular mesangial cells (MC) in the development of diabetic nephropathy (DN). METHODS: Renal biopsy specimens were obtained from patients with type II DN. MC from microdissected glomeruli were cultured in vitro. Both the cells deriving from minute piece of renal biopsy specimen of type 2 DN with overt proteinuria and from transplanted donors' kidneys were investigated. Cell volume, RNA/DNA ratio and the production of fibronectin and glucose transporter 1 (GLUT1) was analyzed by flow ctrometry. Cell proliferation was determined by 3H-Tdr incorporation assay and doubling time. Immunofluorescence staining and flow ctrometry were used to examine the expression of alpha-smooth muscle actin and extracellular matrix. The mRNA expression of GLUT1 was determined by Northern blotting and flow cytometry. Glucose uptake rate by MC was detected using the [3H]-2-DG. The activity of glutamine: fructose-6-P aminotransferase (GFAT), the key enzyme of hexosamine pathway, was measured by spetrophotometry method. Specimens from transplanted kidneys were used a s controls. RESULTS: In specimens from patients with type II DN, increase in volume of MC (in arbitrary units) and RNA/DNA ratio (0.29 +/- 0.05 vs. 0.17 +/- 0.03, P < 0.01) accompanied with high 3H-Tdr incorporation rate (1,898 +/- 421 vs. 1,221 +/- 262, P < 0.05) and shorter doubling time (31.6 +/- 1.84 h vs. 35.0 +/- 5.37 h, P < 0.05) was found in comparison with the control. The synthesis of alpha-smooth muscle actin and extracellular matrix, including fibronectin and laminin, increased. Enhanced mRNA and protein expression of GLUT1 were verified in MC from DN. 2-DG uptake assay showed increased glucose uptake rate in MC from DN compared to the control(1,592 cpm x 10(5) cell-1 vs. 1,275 cpm x 10(5) cell-1, P < 0.05). Furthermore, MC from DN demonstrated a higher GFAT activity as compared to control. CONCLUSION: Dramatic phenotypic and functional changes, including cell hypertrophy, increased cell turnover, excessive formation of ECM, enhanced expression of GLUT1 as well as alterration in cell glucose uptake, excessive flux of glucose metabolism through the hexosamine pathway, occur in MC from type II DN. The above mentioned phenotypic and functional changes may be the basis of pathogenesis of MC in DN. PMID- 11930632 TI - [Diagnostic value of tau in cerebrospinal fluid in alzheimer disease]. AB - OBJECTIVE: To search for reliable and quantitative biochemical marker for diagnosis of Alzheimer disease. METHODS: ELISA-double enzyme amplification assay was used to detect the total tau and abnormally hyperphosphorylated (p-tau) in cerebrospinal fluid specimens of patients with Alzheimer disease (AD, N = 52, 30 from the Netherlands and 22 from Wuhan and Haikou, China), vascular dementia (VD, N = 46, 18 from the Netherlands and 28 from Haikou), and non-neurological disease (N = 37, 13 from the Netherlands and 24 from Haikou) and of normal elderly controls (N = 56, 26 from the Netherlands, and 30 from Wuan and Haikou). RESULTS: The measurement of CSF specimens from Netherlands Brain Bank showed that the levels of total tau and p-tau in CSF specimens of AD patients were significantly higher than those in CSF of patients with VD and non-dementia neurological disorders, and of age-matched non-neurological normal controls. With CSF tau > or = 370 pg/ml as the marker for diagnosis of AD, the sensitivity, specificity, reliability as well as differentiation rate of AD from VD were 90.0%, 79.0%, 82.8% and 66.7%, respectively. With CSF p-tau > or = 120 pg/ml as the marker for diagnosis of AD, the sensitivity, specificity, reliability as well as differentiation rate of AD from VD were 93.3%, 89.5%, 90.8%, and 83.3% respectively. The measurement of total and phosphorylated taus in CSF specimens of AD and non-AD patients and normal controls collected in China showed the similar results; and the sensitivity, specificity, reliability as well as differentiation rate of AD from VD were 77.3%, 85.4%, 83.7% and 71.4% with CSF tau > or = 120 pg/ml as the marker for diagnosis of AD. There was no correlation between CSF tau and p-tau and age, sex, and seriousness of disease. CONCLUSION: Increased phosphorylated tau in human cerebrospinal fluid is a reliable biomarker for diagnosis of Alzheimer disease. PMID- 11930633 TI - [Effects of cyclooxygenase-2 on formation and healing of acetic acid-induced gastric ulcer in rats]. AB - OBJECTIVE: To investigate the possible role of cyclooxygenase-2 (COX-2) inhibitor NS-398 in formation and healing of acetic acid-induced gastric ulcer in rats. METHODS: Fourty eight male Sprague-Dawley rats weighed 160-180 g were perfused with acetic acid into the stomach to induce gastric ulcer and then divided into two groups. NS-398, a specific COX-2 antagonist, was injected subcutaneously 3 hours before and 21 hours after the perfusion with acetic acid and then injected every 24 hours with the dose of 6 mg.kg-1 to 24 rats (treatment group). The other 24 rats were injected subcutaneously with normal saline as controls. In both groups 8 rats were killed 1, 3, and 7 days after the induction of gastric ulcer by acetic acid respectively. RT-PCR and Western blotting were used to determine the expression of COX-2 mRNA and inducible nitric oxide synthase (iNOS) mRNA in the gastric mucosa at different time points. Prostaglandin E2(PGE2) concentration in gastric mucosa was determined by ELISA as a parameter reflecting the COX activity. The severity of ulcer was assessed by ulcer area. RESULTS: COX-2 mRNA expression and PGE2 production were markedly increased in gastric mucosa after ulcer induction, especially in the basal part. After the treatment of NS-398, the increased PGE2 production was inhibited. The ulcer area in NS-398 group was significantly smaller than that in control group 1 day after ulcer induction with slighter congestion and edema around the ulcer. There was no significant difference in ulcer area between NS-398 treatment group and control group 3 days after ulcer induction. However, the ulcer area in NS-398 treatment group was significantly greater than that in control group 7 days after ulcer induction. Along with the severity changes of mucosal lesion, the iNOS expression and activity decreased markedly in the NS-398 group. CONCLUSION: NS-398 inhibits COX 2 activity, thus alleviating inflammatory reaction in acetic acid induced gastric ulcer and averting further damage of tissues. However, it retards the ulcer healing by inhibiting PGE2 production in iNOS expression and activity in gastric mucosa. PMID- 11930634 TI - [Human GDNF cDNA-engineered SH-SY5Y cells' neurotrophic and protective effect on primary dopaminergic neurons of rat]. AB - OBJECTIVE: To construct a kind of engineered cell secreting human GDNF and study its possible effects on gene therapy of Parkinson's disease. METHOD: Human GDNF cDNA with Kozak sequence was cloned by RT-PCR, and then was transfected into SH SY5Y cell line of human neuroblastoma. These engineered cells were co-cultured with primary mesencephalic cells of rats. Dopaminergic neurons were examined by immunohistochemistry. RESULTS: The number of dopaminergic neurons protected by engineered cells increased at least by 95.4% in comparison with the control cells (P < 0.01). The number of dopaminergic neurons protected by engineered cells against MPP+ toxicity increased 9.5-10.8 times (P < 0.01). CONCLUSION: A kind of engineered SH-SY5Y cells secreting human GDNF has been constructed successfully. These cells obviously protect dopaminergic neurons against degeneration and MPP+ toxication and may play an important role in gene therapy of Parkinson's disease. PMID- 11930635 TI - [Intracranial artery occlusive diseases in patients with hypertension and diabetes mellitus]. AB - OBJECTIVES: To investigate risk effects of hypertension (HT) and diabetes mellitus (DM) to intracranial artery occlusive lesions. METHODS: 845 patients (South group) visiting the First Affiliated Hospital of Zhejiang University, and 2325 patients (North group) visiting the Peking Union Medical College Hospital were assessed respeitively. All the patients underwent transcranial Doppler detection to discover intracranial occlusive artery lesions. Age, sex, hypertension (HT) and diabetes mellitus (DM) were included in the analysis. RESULTS: Among the patients > or = 50 years old, the frequencies of middle cerebral artery(MCA) occlusive diseases were: 6.96%, 11.43%, 20.59% and 1.75% in South; 6.24%, 11.18%, 13.51% and 1.95% in North, when suffering from HT, DM, both of HT and DM(MIX), or neither of them(Nor). The risk ratios of HT, DM, MIX were: 4.0, 6.5 and 11.8 in South; 3.2, 5.7 and 6.9 in North; and 2.9, 16.9 and 37.3 when combining South and North. There were no statistical differences of frequencies of MCA occlusive lesions under HT, DM and MIX between the South and North. Multi-variable logistic regression analysis showed that HT and DM were significant and independent predictors for MCA occlusive lesion. The details of other cerebral arteries were canceled here. CONCLUSIONS: Hypertension and diabetes mellitus may contribute to the development of occlusive lesions of the cerebral arteries in Chinese people. PMID- 11930637 TI - [Stem cell transplantation for repairment of injured myocardium]. PMID- 11930636 TI - [Clinical significance of the novel tumor marker CYFRA21-1 in patients with esophageal cancer]. AB - OBJECTIVE: To study the clinical significance of the novel tumor marker--CYFRA21 1 in patients with esophageal cancer. METHODS: The CYFRA21-1 level in serum of 84 patients with a definite diagnosis of esophageal cancer was examined 10 days before and after operation by ELISA. A 3 years' follow-up was conducted to the survival of patients. RESULTS: (1) The CYFRA21-1 level was > 3.3 ng/ml in 72.6% of the patients (61/84). (2) The serum CYFRA21-1 level decreased significantly after operation in patients at stage III or with high differentiation (P < 0.05). (3) The difference between pre- and post-operative serum CYFRA21-1 levels was statistically significant in patients who had undegone radical operation, and was not in patients who had undergone palliative operation. (4) In addition to stage (P < 0.05) and type of operation (P < 0.05), the difference of CYFRA21-1 level before and after operationwas closely related to the prognosis (P < 0.05). CONCLUSION: CYFRA21-1 is a useful marker in diagnosis and prediction of prognosis of esophageal cancer. PMID- 11930639 TI - Roles of NHE-1 in the proliferation and apoptosis of pulmonary artery smooth muscle cells in rats. AB - OBJECTIVE: To evaluate the roles of Na+/H+ exchanger-1 (NHE-1) in the proliferation and apoptosis of pulmonary artery smooth muscle cells in rats. METHODS: Twenty Wistar rats were randomized into control group and 3-week hypoxic group. Intracellular pH (pHi) of the smooth muscle was determined with fluorescence measurement of the pH-sensitive dye BCECF-AM, and the expression of NHE-1 mRNA was detected by reverse transcription polymerase chain reaction (RT PCR). Primary culture of pulmonary artery smooth muscle cells in vitro was performed. In situ cell death detection kit (TUNEL) was used for studying the effect of specific NHE-1 inhibitor-dimethyl amiloride (DMA) on the apoptosis of muscle cells which had intracellular acidification. RESULTS: pHi value and NHE-1 mRNA expression of pulmonary artery smooth muscle cells were significantly higher in the hypoxic group than in the control group (P < 0.01, P < 0.001). DMA elevated the apoptotic ratio remarkably. The effect was enhanced when DMA concentration increased and the time prolonged. CONCLUSIONS: With the function of adjusting pHi, NHE-1 may play an important role in the proliferation and apoptosis of pulmonary artery smooth muscle cells. PMID- 11930638 TI - Effect of trans-acting factor on rat glutathione S-transferase P1 gene transcription regulation in tumor cells. AB - OBJECTIVE: To investigate the effect of trans-acting factor(s) on rat glutathione S-transferase P1 gene (rGSTP1) transcription regulation in tumor cells. METHODS: The binding of trans-acting factor(s) to two enhancers of the rGSTP1 gene, glutathione S-transferase P enhancer I (GPEI) and glutathione S-transferase P enhancer II-1 (GPE II-1), was identified by an electrophoretic mobility shift assay (EMSA). The molecular weight of trans-acting factor was measured in a UV cross-linking experiment. RESULTS: Trans-acting factor interacting with the core sequence of GPEI (cGPEI) were found in human cervical adenocarcinoma cell line (HeLa) and rat hepatoma cell line (CBRH7919). These proteins were not expressed in normal rat liver. Although specific binding proteins that bound to GPE II-1 were detected in all three cell types, a 64 kDa binding protein that exists in HeLa and CBRH7919 cells was absent in normal rat liver. CONCLUSION: cGPEI, GPEII specific binding proteins expressed in HeLa and CBRH7919 cells may play an important role in the high transcriptional level of the rGSTP1 gene in tumor cells. PMID- 11930640 TI - Effect of a cyclosporine A delivery system in corneal transplantation. AB - OBJECTIVE: To test the immunosuppressive effect of cyclosporine (Cs) in a polymer placed in the anterior chamber of corneal allograft recipients. METHODS: Wistar inbred rats with vascularized corneas were recipients of corneal allografts from Sprague-Dawley donor rats. Rats underwent penetrating keratoplasty and were divided randomly into four groups: untreated control animals (UCA); Cs-polymer anterior chamber recipients (CPA); co-polymer subconjunctival recipients (CPS); and Cs-olive oil drop recipients (COO). Grafts were examined by slit lamp every 3 days and clinical conditions were scored. Cs concentration in the aqueous humor was assayed at 1, 2, and 4 weeks. At 1, 2 and 4 weeks after transplantation, the operated eyes were collected for histopathological evaluation of the grafts. RESULTS: The median survival time of the allografts was 8.2 +/- 1.48 days for the UCA group, 11.4 +/- 2.50 days for the CPS group, and 17.0 +/- 2.00 days for the CPA group. There was a statistically significant difference (P < 0.05) between survival time of the allografts in the animals of the CPA group compared to the other groups of graft recipients. Significantly higher concentrations of Cs were found in the eyes given an anterior chamber implant of Cs-polymer, compared to other treatment groups or untreated rats. A transient inflammatory response in the anterior chamber was observed in the CPA group. CONCLUSIONS: Cs-polymer placed in the anterior chamber significantly prolongs corneal allograft survival time in a high risk corneal graft rejection model. This intraocular delivery system may be a valuable adjunct for the suppression of immune graft rejection. PMID- 11930641 TI - Use of cutting balloons in coronary interventions. AB - OBJECTIVE: To study the efficacy and safety of cutting balloons in coronary interventions. METHODS: Twenty-two patients with 23 narrowed coronary arteries and 25 lesions enrolled the study, 17 of whom had 18 diseased coronary arteries and 18 lesions were in-stent restenosis. The average time from previous stenting was 7.60 +/- 3.53 months. The lesions were dilated with cutting balloons. RESULTS: All of the lesions were dilated successfully by 5.16 +/- 2.30 inflations of a cutting balloon. The mean total duration of balloon inflation was 233.96 +/- 94.83 seconds at pressures up to 9.40 +/- 1.96 bars. The severity of vascular stenosis was lessened substantially (89.64 +/- 8.65% vs 17.60 +/- 17.15%, P = 0.000) without severe complications. Three restenotic lesions were further dilated with conventional balloons, another one was stented again because of a dissection distal to the previous stent. Three primary lesions were stented for dissection or residual stenosis. Angina pectoris reoccurred in two patients in a mean follow-up period of 7.42 +/- 6.87 (range 0.5-20) months. CONCLUSION: Cutting balloon dilation is an effective and safe choice in interventions for coronary disease especially for in-stent restenosis. PMID- 11930643 TI - Prevalence and incidence of urinary incontinence in the elderly: a longitudinal study in South Australia. AB - OBJECTIVE: To identify the prevalence, incidence and trend of urinary incontinence in the elderly over a two-year period. METHODS: We randomly selected 4187 elderly people using the State Electoral Data Base in South Australia. Questionnaire surveys were conducted in 1992 with a response rate of 53.4%. The following two surveys were conducted 12 and 24 months later. A total of 2087 elderly people completed all three surveys. RESULTS: Urge incontinence had a higher prevalence rate, with 36.6% to 41.6% of the elderly having at least an occasional problem (with 7.5% to 9.6% having problems often), compared to 23.4% to 28.8% having at least an occasional problem with stress incontinence (with 3.3% to 5.0% having problems often). Combined incontinence and urge incontinence were more common than stress incontinence alone. The incidence rates of urge incontinence and stress incontinence were 19.8% (with 3.1% often) and 14.5% (with 1.9% often), respectively. The trends toward incontinence during the two-year period indicated increasing trends in the elderly. CONCLUSION: Urinary incontinence is an important health problem in the elderly and our data show that it has an increasing trend for future. PMID- 11930642 TI - Direct gastroscopy for detecting gastric cancer in the elderly. AB - OBJECTIVE: To evaluate the safety and effectiveness of direct gastroscopy for detecting gastric cancer. METHODS: Clinical screening by direct gastroscopy was performed for gastric cancer (GC) from September 1985 to July 1998. 3048 elderly people were screened. Their age ranged from 60 to 93 years, and 2034 of the 3084 were followed up. RESULTS: Ninety-two patients with gastric cancer were discovered by gastroscopy, representing 3.02% of the screened population. The rate of early gastric cancer (EGC) was 63.04% (58/92) of all gastric cancers detected. The rate was up to 79.59% (39/49) on follow-up, and was 74.14% (43/51) in asymptomatic patients with gastric cancer. The excision rate was 88.89% for patients with gastric cancer, and 100% for patients with early gastric cancer. The 5-year survival rate was 91.89% for patients with gastric cancer, and 96.30% for patients with early gastric cancer. CONCLUSION: Clinical screening and follow up by direct gastroscopy in persons over 60 years of age are a safe and effective method for raising the 5-year survival and detection rate of gastric cancer, especially early gastric cancer. PMID- 11930644 TI - Correlation of expression of preprothyrotropin-releasing hormone and receptor with rat testis development. AB - OBJECTIVE: To investigate the expression regulation of thyrotrophin-releasing hormone (TRH) and TRH receptor (TRH-R), and their role in the development of rat testis. METHODS: Oligonucleotide primers were designed from the sequences of rat hypothalamus prepro TRH (ppTRH) and pituitary TRH-R cDNA for reverse transcription polymerase chain reaction (RT-PCR). Specific fragments of ppTRH and TRH-R cDNA were cloned and sequenced. Expression plasmids containing ppTRH and TRH-R genes were then constructed, and expression was found in E. coli DH5-alpha. ppTRH and TRH-R mRNA in the testis was quantitated in RNA samples prepared from rats at different developmental stages by real time quantitative RT-PCR. RESULTS: The quantitative analyses demonstrated that no ppTRH and TRH mRNA could be detected at the earliest stage (day 8). ppTRH and TRH mRNA signals were detected on day 15 and increased progressively on days 20, 35, 60 and 90. CONCLUSION: Our results suggest that rat testis could specifically express TRH and TRH-R, and the transcriptions of ppTRH and TRH-R genes in the rat testis were development dependent. The acquirement of expressed products for ppTRH and TRH-R can be used for further research on the physiological significance of TRH and TRH-R expression in rat testis. PMID- 11930645 TI - Evaluation of renal function in children with moderate or severe hydronephrosis after operation. AB - OBJECTIVE: To evaluate postoperative renal function in children with congenital moderate or severe hydronephrosis. METHODS: 99mTc-labeled diethylenetriaminepenta acetic acid scintigraphy was performed in 50 children with unilateral moderate or severe hydronephrosis to determine postoperative renal function. We also analyzed the factors influencing renal function recovery. RESULTS: Average postoperative renal function in 50 cases was 40.62% +/- 10.09%. Among them, 32% of patients had nearly normal renal function and differentiated renal function reached up to 45%. Average preoperative and postoperative renal function in 25 cases was 23.89% +/- 11.65% and 39.33% +/- 8.59% respectively and the increase of renal function was about 15.44% +/- 11.18% (P = 0.0003). Renal parenchyma thickness was negatively correlated with postoperative renal function (r = -0.62, P = 0.0009). The follow up period was positively correlated with postoperative renal function (r = 0.58, P = 0.0026). The patients' age had no correlation with renal function recovery (r = -0.05, P = 0.80). Recovery of renal function in hydronephrosis with extrarenal pelvis was greater than that in hydronephrosis with intrarenal pelvis (P = 0.016). CONCLUSIONS: Postoperative renal function in children with moderate or severe hydronephrosis can recover to normal. Recovery of renal function was more obvious in hydronephrosis with thinner renal parenchyma, longer follow-up period and extrarenal pelvis. PMID- 11930646 TI - Transcatheter closure of secundum atrial septal defect using an Amplatzer septal occluder. AB - OBJECTIVE: To evaluate the safety and efficacy of the Amplatzer septal occluder for transcatheter closure in patients with secundum atrial septal defect (ASD II). METHODS: Patients with clinically confirmed ASD II were recommended for transcatheter closure of ASD II. RESULTS: 30 ASD II patients (20 females) underwent transcatheter closure at a median age of 18.4 years (5-55 years). Both the stretched diameters of ASDs and the sizes of the devices were from 18 to 34 mm (25 +/- 7 mm). The successful placement rate was 100%. The rest shunt documented by color Doppler, was immediately after implantation in 40% of patients, in 9.9% after 24 hours, and in 3.3% trace at 3 months. No serious complications were observed. There was improvement in symptoms and in cardiac size. Septal motion abnormalities normalized in all patients after 3 months follow-up. CONCLUSION: The Amplatzer septal occluder is a safe and effective device for transcatheter closure of ASD II. Long-term follow-up is still required before widespread clinical use can be recommended. PMID- 11930647 TI - Management of nephropathy in patients with type 2 diabetes. AB - PURPOSE: To review evidence-based management of nephropathy in patients with type 2 diabetes. DATA SOURCES: A literature search (MEDLINE 1966 to 2000) was performed using the key word "diabetic nephropathy". Relevant book chapters were also reviewed. STUDY SELECTION: Well-controlled, prospective landmark studies and expert review articles on diabetic nephropathy were selected. DATA EXTRACTION: Data and conclusions from the selected articles that provide solid evidence to the optimal management of diabetic nephropathy were extracted and interpreted in light of our clinical research experience with many thousands of Hong Kong Chinese patients. RESULTS: Hypertension, long diabetes duration, poor glycaemic control and central obesity are the most important risk factors. Microalbuminuria is a practical marker to predict overt nephropathy in type 2 diabetic patients. Risk factor modification, renal function monitoring and combined therapies are the current integrated approaches to manage patients with diabetic kidney disease. Optimal glycaemic control is the mainstay of treatment but effective antihypertensive therapy is also key to delaying the progression of diabetic nephropathy. Angiotensin-converting enzyme inhibitors and angiotensin II receptor antagonists have important renoprotective actions independent of their blood pressure lowering actions. CONCLUSIONS: Diabetic nephropathy is the leading cause of end-stage renal disease worldwide. Monitoring renal function and screening for microalbuminuria will allow the identification of patients with nephropathy at a very early stage for intervention. Tight glycaemic control and aggressive antihypertensive treatment as well as the use of renin-angiotensin system inhibitors should substantially delay the progression of nephropathy. PMID- 11930648 TI - Nasopharyngeal carcinoma with particular characteristics: case report. PMID- 11930649 TI - Primary omental malignant mixed mullerian tumor in a 67-year-old woman. PMID- 11930650 TI - Lead poisoning in new immigrant children from the mainland of China. AB - OBJECTIVE: To define the prevalence, severity and risk factors for lead poisoning in new immigrant children from the mainland of China to Hong Kong, China. METHODS: New immigrant children from the mainland of China under 18 years of age were invited to join the study. Their growth parameters and venous blood lead levels (BLL) were measured within 7 days of arrival. Those with elevated BLL i.e. > 10 micrograms/dl (0.48 mumol/L) were assessed for signs, symptoms and risk factors of lead poisoning. Education on the prevention of lead poisoning and follow up BLL measurements were offered until their BLL normalized. RESULTS: Four hundred and fifty-seven children were recruited. Among them, 18.1% and 2.6% had BLL > 0.48 and 0.71 mumol/L, respectively. None had BLL > 0.96 mumol/L. Possible risk factors included contaminated drinking water (19%), herb ingestion (17.5%), pica, playing in dumping grounds, residing near paint factories or highways, habitual inhalation of car exhaust and cooking with petroleum. Symptoms and signs included abdominal pain, headache, short stature, and learning difficulties but did not correlate with BLL. None required treatment except for counseling on the avoidance of risk factors. About 94.7% showed a reduction in BLL 2-3 months after arrival and before counseling. All had normalized BLL by 9 months except 1 who had an X-ray feature of lead line. CONCLUSIONS: Lead poisoning is common in new immigrant children from the mainland of China. Environmental factors were most important while behavioral factors like regular herb ingestion might have contributed to the persistence of high BLL in the minority. PMID- 11930651 TI - Expression of P16 protein and Bcl-2 protein in malignant eyelid tumors. AB - OBJECTIVE: To investigate the relationship between P16 gene (the tumor suppressor gene) and the bcl-2 gene (the apoptosis inhibitor gene) and the incidence and development of malignant eyelid tumors. METHODS: The streptavidin-biotin peroxidase complex immunohistochemistry method was used to study the expression of P16 gene and the bcl-2 gene in 96 cases of malignant eyelid tumors. RESULTS: Among the 96 cases, there were 40 basal cell carcinomas (BCCs), 33 squamous carcinomas and 23 sebaceous carcinoma, with P16 protein positive (nuclear staining) rates 70%, 54.6% and 56.5%, respectively. The P16 positive rate was negatively correlated with the degree of tumor histological differentiation, and the rate difference between the high differentiated carcinomas was significant (P < 0.05). Positive Bcl-2 protein expression was detected in the cytoplasm. All 40 BCC cases were Bcl-2 positive, and nearly all of the tumor cells showed positive cytoplasmic expression, while in the 33 squamous cell carcinoma cases only one showed positive focal reaction, and the staining in the other 32 cases was relatively faint. None of the 23 sebaceous carcinomas expressed Bcl-2. CONCLUSIONS: The expression of the P16 protein was related to the occurrence and degree of differentiation of malignant eyelid tumors. The overexpression of the Bcl-2 protein suggests that suppression of apoptosis might play a role in the tumorigenesis of BCC. PMID- 11930652 TI - Percutaneous transluminal septal myocardial ablation in patients with hypertrophic obstructive cardiomyopathy. AB - OBJECTIVE: To evaluate the immediate and follow-up results of percutaneous transluminal septal myocardial ablation (PTSMA) in patients with hypertrophic obstructive cardiomyopathy (HOCM). METHODS: Fifteen symptomatic, drug-refractory patients with HOCM underwent PTSMA procedures with application of a myocardial contrast echocardiography (MCE) intra-procedure. Before and after the procedure, clinical evaluations were obtained in all patients, who were followed up for a mean period of 8.6 +/- 3.8 (6-20) months. RESULTS: Immediate left ventricular outflow tract gradient (LVOTG) reduction was achieved (77.93 +/- 22 mm Hg vs 14.8 +/- 15 mm Hg, P < 0.0001) after the procedure with a mean decrease of 5.75 +/- 2.87 mm Hg of left ventricular end diastolic pressure (P < 0.001). Follow up results revealed that ventricular remodelling occurred mainly 1-3 months after the procedure, but without evidence of ventricular dilation and contract dysfunction. Heart function (NYHA) was greatly improved (3.4 +/- 0.5 vs 1.1 +/- 0.4, P < 0.001) and exercise endurance increased. A renewed increase of LVOTG was found in 2 patients during follow-up. CONCLUSIONS: LVOTG was greatly decreased in HOCM patients undergoing a PTSMA procedure, and their symptoms were greatly improved without cardiac complications during follow-up. Sub-selection and reopening of target vessels were the causes of renewed increase of LVOTG, and this can be avoided with the accumulation of experience. This is a promising method for the treatment of symptomatic patients with HOCM. PMID- 11930653 TI - Clinical and laboratory features of preleukemia patients. AB - OBJECTIVE: To explore prospective diagnostic criteria for preleukemia. METHODS: A case control study was done comparing the discrepancies on clinical and laboratory features between patients with preleukemia and those with chronic aplastic anemia (CAA) or atypical paroxysmal nocturnal hemoglubinuria (a-PNH). RESULTS: There were eight variables of significance: (1) lymphocytoid micromegakaryocytes in the bone marrow; (2) immature granulocytes in the peripheral blood; (3) > or = 2.0% myeloblasts in the bone marrow; (4) positive periodic acid schiff (PAS) stained nucleated erythrocytes; (5) myeloid differentiation index > or = 1.8; (6) typical colonal karyotypic abnormalities; (7) negative sister chromatid differentiation; (8) cluster/colony ratio of granulocyte-macrophage colony-forming units (CFU-GM) > 4.0. The following criteria were assigned: A: to meet variable one and at least two of the other seven variables and B: to meet at least four of the eight variables. All of the patients with preleukemia met either A or B and none of the patients with CAA or a-PNH did. CONCLUSIONS: Preleukemia is different from CAA or a-PNH. It has its own clinical and laboratory features, which may be useful for its prospective diagnosis. PMID- 11930654 TI - Accumulation of ciprofloxacin and lomefloxacinin fluoroquinolone-resistant strains of Escherichia coli. AB - OBJECTIVE: To evaluate the role of outer membrane protein (Omp) F-deficiency and active efflux in the accumulation of hydrophilic fluoroquinolones ciprofloxacin (CPLX) and lomefloxacin (LMLX) in resistant E. coli strains. METHODS: Fluoroquinolone accumulation in bacteria and the effect of active efflux were measured by a fluorescence method. The outer membrane proteins of the bacteria were analysed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS PAGE). E. coli strains in this study included control strains JF701 and JF703 that are OmpC- or OmpF-deficient mutants of E. coli K-12, respectively, and the fluoroquinolone susceptible strain the fluoroquinolone susceptible strain of Escherichia coli (Ecs) and its in vitroselected resistant strains R2 and R256, and the clinical resistant isolates R5 and R6. RESULTS: The steady-state accumulation concentration of each drug in Ecs appeared to be the same as in JF701, while in the OmpF-deficient strain JF703, it was 1/5 CPLX or 1/2 LMLX lower than that in JF701, but JF703 was still susceptible to fluoroquinolones. On the other hand, compared with susceptible strains, a 2- to 10-fold decrease in the accumulation of each drug was found in the resistant strains except R2, in which the accumulation was slightly higher than in JF703. After the addition of 2,4-dinitrophenol (DNP), accumulation of each drug increased, especially in resistant strains, indicating that the function of the active efflux (pump) system in these bacteria had been enhanced dramatically. Furthermore, both OmpF and OmpC in Ecs, OmpF-deficiency in R2 and R256 and OmpC-deficiency in R5 and R6 were observed. CONCLUSION: The decreased accumulation of hydrophilic fluoroquinolones in E. coli involved OmpF-deficiency and active efflux (pump), and the latter may be an important factor. PMID- 11930655 TI - Gene expression profile differences in high and low metastatic human ovarian cancer cell lines by gene chip. AB - OBJECTIVES: To study the difference between gene expressions of high (H0-8910PM) and low (HO-8910) metastatic human ovarian carcinoma cell lines and screen novel associated genes by cDNA microarray. METHODS: cDNA retro-transcribed from equal quantities of mRNA derived from high and low metastatic tumor cells or normal ovarian tissues were labeled with Cy5 and Cy3 fluorescein as probes. The mixed probe was hybridized with two pieces of BioDoor 4096 double dot human whole gene chip and scanned with a ScanArray 3000 laser scanner. The acquired image was analyzed by ImaGene 3.0 software. RESULTS: A total of 355 genes with expression levels more than 3 times larger were found by comparing the HO-8910 cell with normal ovarian epithelial cells. A total of 323 genes with expression levels more than 3 times larger in HO-8910PM cells compared to normal ovarian epithelium cells were also detected. A total of 165 genes whose expression levels were more than two times those of HO-8910PM cells compared to their mother cell line (HO 8910) were detected. Twenty-one genes with expression levels > 3 times were found from comparison of these two tumor cell lines. CONCLUSIONS: cDNA microarray techniques are effective in screening differential gene expression between two human ovarian cancer cell lines (H0-8910PM; HO-8910) and normal ovarian epithelial cells. These genes may be related to the genesis and development of ovarian carcinoma. Analysis of the human ovarian cancer gene expression profile with cDNA microarray may help in gene diagnosis, treatment and prevention. PMID- 11930656 TI - Usefulness of determining a protein induced by vitamin K absence in detection of hepatocellular carcinoma. AB - OBJECTIVE: Protein induced by vitamin K absence or antagonist II (PIVKA II), also called des-gamma carboxy prothrombin (DCP), is a sensitive marker for the diagnosis of hepatocellular carcinoma (HCC), in Japan and the United States since the sensitive kits were available (1998). PIVKA II is not used in clinical diagnosis in China so far. The aim of this study was to assess the diagnostic value of PIVKA II in Chinese patients with HCC. METHODS: Serum PIVKA II and alpha fetoprotein (AFP) levels were determined in 60 patients with HCC and 30 patients with cirrhosis not carrying HCC. RESULTS: The mean serum concentration of PIVKA II in HCC patients (784.3 +/- 1364.1 mean +/- s) was higher than that in cirrhosis patients (16.1 +/- 31.7); this difference was highly significant (P < 0.0001). When the cutoff level of 40 mAU/ml was used as the level of discriminating HCC from cirrhosis, 51.7% of patients (31/60) with HCC had PIVKA II values above this level (sensitivity). Only 4 patients with cirrhosis had such high PIVKA II levels. Thus, the specificity of this test was 86.7% (26/30). Total accuracy was 62.2% [(31 + 26)/(60 + 30)]. Seven of 19 small HCCs (36.84%) had PIVKA II values above the cutoff level. Concentrations of AFP above 20 ng/ml were observed in 34 of 60 patients with HCC (56.7%) and in 11 patients with cirrhosis (36.7%). Eleven of 26 patients with HCC (46.2%) without increased AFP had concentrations of PIVKA II greater than 40 mAU/ml. No significant correlation was found between serum levels of AFP and PIVKA II that were measured in 60 HCC patients (rs = 0.101, P = 0.247). Combining the information from PIVKA II and AFP showed an increase of approximately 21.6% over AFP and 26.7% over PIVKA II alone. For small HCC patients, combining the information from PIVKA II and AFP showed an increase of approximately 15.8% over AFP alone and 21.1% over PIVKA II alone. CONCLUSION: PIVKA II is a useful early diagnostic marker for HCC and may be more sensitive when combined with AFP in Chinese patients. PMID- 11930657 TI - Interleukin-1 gene polymorphism disease activity and bone mineral metabolism in rheumatoid arthritis. AB - OBJECTIVE: To determine whether interleukin-1 alpha and 1 beta gene polymorphism is associated with rheumatoid arthritis disease activity and bone mineral metabolism, and whether there is any relationship between IL-1 beta and rheumatoid arthritis (RA) motif gene. METHODS: IL-1 gene polymorphisms were analyzed in 65 RA patients who met American College of Radiology (ACR) criteria and 60 controls. From genomic DNA, 2 polymorphisms in each gene for IL1 alpha-889 and IL-1 beta + 3953 were typed by PCR-RFLP and HLA-DRB1 allele typing was also undertaken by PCR-SSOP. Some clinical and laboratory parameters were collected. The allelic frequencies and carriage rates were compared between RA patients and controls and between patients with active and quiescent disease. Comparison was also made between IL-1 polymorphism and parameters of bone mineral metabolism and between patients with the HLA-DRB1 RA motif plus IL-1 beta 2 and patients without the two alleles. Fisher test and the analysis of variance was used to analyze the data. RESULTS: There was no significant difference in the frequency and carriage rate of IL-1 alpha polymorphisms between RA patients and the controls. The beta 2/2 genotype of IL-1 beta was more common in female RA patients compared with controls (P = 0.001). A lower carriage rate of IL-1 beta 2 occurred in male RA patients (P = 0.001). A higher carriage rate of IL-1 alpha 2 is associated with a higher ESR (P = 0.008), HAQ score (P = 0.03), and vit-D3 (P < 0.001), but conversely a lower SJC (p = 0.002), a lower RF (P = 0.002) and a lower BMD at the lumbar spine (P = 0.001). A higher frequency of IL-1 alpha 1 is associated with a lower CRP value (P = 0.009). An increased IL-1 beta 2 carriage is associated with active rheumatoid disease as indicated by a higher CRP (P < 0.001), ESR (P < 0.001) and pain score (P = 0.001) and a higher BMD at the lumbar spine (P = 0.007), lower vit-D3 and. Udpd/Crea level The presence of the HLA DRB1 RA motif and IL-1 beta allele 2 at same time did not contribute to disease activity. CONCLUSION: Polymorphisms of the IL-beta gene may affect the RA occurrence. Carriage of IL-1 beta 2 polymorphisms is associated with more active disease in RA and the presence of both the IL-1 alpha 2 and the IL-1 beta 1 allele in RA influences bone resorption. PMID- 11930658 TI - Normative values of pulmonary function testing in Chinese adults. AB - OBJECTIVES: To compare the difference in pulmonary function between Caucasians and Chinese and assess the best reference values of pulmonary function for Chinese adults. METHODS: Values for pulmonary function were predicted by tests on 4773 Chinese healthy subjects (male: 2560, female: 2213, aged 15-78 years) in six parts (north, northeast, northwest, east, southwest and south) of China. Prediction equations of the European Community for Steel and Coal (ECSC), other equations for overseas Chinese or for Caucasians were also selected. The regression coefficients of forced vital capacity (FVC), forced expiratory volume in one second (FEV1), FEV1/FVC, total lung capacity (TLC) and residual volume (RV) were summarized. RESULTS: ECSC predictions were closer to the Chinese ones than other selected equations. Comparison with ECSC predictions showed that on average the values for FVC, FEV1 were 5.3% smaller in Chinese males and 3.3% smaller in Chinese females, with the maximal differences in south China and the minimal differences in North China. RV and TLC in Chinese were lower than in Caucasians (males 4.8%, 5.5%, respectively; females 8.7% and 6.0%, respectively). Conversion factors were given for adjusting ECSC equations to fit Chinese. CONCLUSIONS: For predicting values of pulmonary function in Chinese, we suggest to use the equations reported here. Alternatively, ECSC regression equations may be used with appropriate conversion factors. PMID- 11930659 TI - Coronary bypass revascularization with radial artery and internal mammary artery grafts. AB - OBJECTIVE: To evaluate radial artery (RA) and internal mammary artery (IMA) grafts in coronary artery bypass and the use of color Doppler ultrasound in the peri-operative evaluation of IMA and radial-ulnar collateral circulation. METHODS: From June 1998 to June 2000, sixty cases of coronary bypass revascularization with RA and IMA were performed. Preoperatively, the radial ulnar collateral circulation was evaluated with the modified Allen's test, color Doppler ultrasound and noninvasive oxygen saturation measurement. The IMA lumen and blood flow were measured at the first intercostal space with color Doppler ultrasound preoperatively and postoperatively. RESULTS: One patient (1.7%) died of serious cardiac arrhythmia on the fourth postoperative day. There were no arterial graft harvest related complications. Before harvesting, the ulnar artery blood flow was 30.78 +/- 9.71 ml/min, and it increased to 43.36 +/- 13.98 ml/min (40.87% increase, P < 0.01) after the operation. Compared with the baseline, there was no obvious change of IMA blood flow postoperatively (P > 0.05), but the systolic/diastolic flow ratio markedly decreased from 8.57 +/- 3.98 ml/min to 3.41 +/- 4.87 ml/min (P < 0.01). CONCLUSIONS: Arterial grafts can be safely used for coronary bypass revascularization with good results. The ulnar artery blood flow can increase compensatively after RA harvesting. The diastolic blood flow of grafted IMA markedly increased postoperatively. Color Doppler ultrasound was very helpful both in evaluating the radial-ulnar collateral circulation before RA harvesting and in assessing the patency of the grafted IMA after coronary artery bypass grafting (CABG). PMID- 11930660 TI - All-trans retinoic acid as a single agent induces complete remission in a patient with acute leukemia of M2a subtype. AB - OBJECTIVE: To present a special case with the karyotype and molecular marker of acute myeloid leukemia (AML)-M2 who was induced to complete remission by all trans retinoic acid (ATRA) alone. METHODS: A recently hospitalized young female patient with acute leukemia was initially diagnosed as M3 subtype based on morphological French-American-British (FAB) classification. Karyotype analysis using standard G and R banding techniques and RT-PCR were applied to further define the diagnosis. After primarily cultured bone marrow cells from the iliac aspiration were tested for in vitro induced differentiation, the patient was treated with oral all-trans retinoic acid alone, 60 mg per day until complete remission was achieved. Peripheral blood and bone marrow changes were monitored over the whole treatment course. RESULTS: The characteristic chromosomal aberration for M3, the t(15;17) reciprocal translocation, was not found while a t(8;21) translocation was verified. Furthermore, an amplified product of the AML 1/ETO fusion gene instead of the PML/RAR alpha fusion gene was detected by RT-PCR and the diagnosis was corrected from M3 to M2. Primary cultured bone marrow cells can be fully induced to terminal differentiation after 4 days exposure to ATRA. A hematological complete remission was achieved after 40 days treatment with ATRA as a single therapeutic agent, suggesting an alternative pathway mediating ATRA induced myeloid differentiation. CONCLUSION: A leukemia patient with a subtype other than M3, such as M2 in this case, may also be induced to complete remission by the mechanism of ATRA-induced terminal differentiation. This implies that there may be a pathway other than PML/RAR alpha fusion gene product which mediates ATRA-induced myeloid maturation in leukemia cells. PMID- 11930662 TI - Hepatic microcirculatory disturbances in patients with chronic hepatitis B. AB - OBJECTIVE: To document morphological changes in hepatic microcirculation in liver tissue with hepatitis B and the pathogenesis of hepatic microcirculatory disturbances. METHODS: Liver tissue samples were obtained from patients with hepatitis B by liver biopsy. These samples were examined with a light microscope and transmission electron microscope. RESULTS: Hepatic microcirculatory disturbances existed in patients with hepatitis B, including those with normal liver function, manifested by red blood cell aggregation in sinusoids seen under light microscope and sinusoidal capillarization seen under electron microscope. Weibel-Palade bodies in sinusoidal endothelial cells were seen in 26 out of 53 cases. Intimate contacts were found between lymphocyte/Kupffer cells and sinusoidal endothelial cells. CONCLUSIONS: Hepatic microcirculatory disturbances exist in patients with hepatitis B. The appearance of Weibel-Palade bodies in sinusoidal endothelial cells may be a key step in the development of hepatic microcirculatory disturbances. PMID- 11930661 TI - Down stream involvement of the bile duct in hepatolithiasis. AB - OBJECTIVE: To evaluate the down stream involvement of the bile duct in hepatolithiasis. METHODS: Mechanical damage to bile duct epithelia and long standing cholangitis as result of hepatolithiasis play an important role in the carcinogenesis of bile duct epithelia and stricture of the intra- and extra hepatic bile duct. Macromorphological and microscopic changes in bile duct mucosa of 100 consecutive patients with hepatolithiasis were investigated using intra- or post-operative cholangioscopy. Biopsy specimens of lesions obtained during cholangioscopy were studied with immunohistochemical staining and flow cytometry to determine proliferative activity and DNA content. Five cases of well-proven cholangiocarcinoma were simultaneously studied as controls. RESULTS: Of the 100 patients, those with chronic cholangitis accounted for 86% (86/100), proliferative lesions 11% (11/100), adenomatous polyps 1% (1/100), and adenocarcinoma 2% (2/100). The obvious mucosal lesion associated with hepatolithiasis was located down-stream of the bile duct, predominantly in the hilar region, e.g. orifices of the right/left hepatic duct and common hepatic duct (73% mucosa lesions in the hilar region). The intensity of cancer embryonic antigen stain and the proliferative cell nuclear antigen index increased with the development of bile duct lesions. Aneuploid DNA presented mainly in the high degree malignant adenocarcinomas (> 80% of cases). CONCLUSIONS: The obvious mucosal lesions associated with hepatolithiasis were located down-stream of the bile duct, predominantly in the hilar region (73% of mucosal lesions). The proliferative activity of examined bile duct mucosa lesions increased with the development of pathological deterioration, which may contribute to the development of hilar bile duct stricture and hilar cholangiocarcinoma. PMID- 11930663 TI - Clonal expansion T cells identified in acute monoblastic leukemia by CDR3 size analysis of TCR V beta repertoire using RT-PCR and genescan. AB - OBJECTIVE: To investigate the distribution and clonality of T cell receptor (TCR) V beta repertoire in patients with acute monoblastic leukemia (AML-M5). METHODS: Expression of the TCR V beta repertoire was analyzed using reverse transcription polymerase chain reaction (RT-PCR), which amplified the complementarity determining region 3 of 24 TCR V beta genes in peripheral blood from 9 cases with acute myclogenous leukemia subtype 5 or acute monoblastic leukemia (AML-M5). PCR products were further studied by genescan analysis to identify T cell clonality. RESULTS: Expression of 1-10 V beta subfamilies was found in samples from 9 patients. Genescan analysis showed that some V beta subfamily products from 8 of 9 cases contained an oligoclonal peak. Oligoclonal T cells of the V beta 2 subfamily could be found in 6 patients with AML-M5. CONCLUSIONS: T cell clonality expansion was found in AML-M5 cases and were tendentious in the V beta 2 subfamily, suggesting a the specific immune response for leukemia cell (M5) associated antigen and may display antileukemia activity. PMID- 11930664 TI - Relationship between microdeletion on Y chromosome and patients with idiopathic azoospermia and severe oligozoospermia in the Chinese. AB - OBJECTIVES: To evaluate the relationship between microdeletion or mutation on the Y chromosome and Chinese patients with idiopathic azoospermia and severe oligozoospermia and to establish a molecular detection method. METHODS: Microdeletion or mutation detection at the AZFa (sY84 and USP9Y), AZFb, AZFc/DAZ and SRY regions of the Y chromosome. Seventy-three azoospermia and 28 severe oligozoospermia patients were evaluated using PCR and PCR-SSCP techniques. RESULTS: Twelve of 101 patients (12%) with the AZFc/DAZ microdeletion were found, including 8 with azoospermia (11%) and 4 with severe oligozoospermia (14.3%), and 1 patient had a AZFb and AZFc/DAZ double deletion. No deletions in the AZFa or SRY regions were found. No deletions in AZFa, AZFb, AZFc/DAZ or SRY regions were found in 60 normal men who had produced one or more children. CONCLUSIONS: Microdeletion on the Y chromosome, especially at its AZFc/DAZ regions, may be a major cause of azoospermia and severe oligozoospermia leading to male infertility in China. It is recommended that patients have genetic counseling and microdeletion detection on the Y chromosome before intracytoplasmic sperm injection. PMID- 11930665 TI - Analysis of the GM-CSF and GM-CSF/IL-3/IL-5 receptor common beta chain in a patient with pulmonary alveolar proteinosis. AB - OBJECTIVE: To investigate the expression of the granulocyte-macrophage colony stimulating factor (GM-CSF) and GM-CSF/IL-3/IL-5 receptor common beta chain (beta c receptor) in an adult patient with idiopathic pulmonary alveolar proteinosis (PAP), so as to demonstrate the possible association of the GM-CSF and beta c receptor with the pathogenesis of human PAP. METHODS: The GM-CSF levels were measured with a commercial ELISA kit (sensitivity 5 pg/ml) and the beta c receptor expression on the cell surface was detected by flow cytometry analysis. Reverse transcription polymerase chain reaction (RT-PCR) analysis was employed to detect the expression of the GM-CSF mRNA and the beta c receptor mRNA in peripheral blood mononuclear cells and alveolar macrophages. The entire coding regions of the GM-CSF cDNA and the beta c receptor cDNA were sequenced by the Sanger dideoxy-mediated chain termination method to detect possible mutations. RESULTS: The patient with PAP failed to release the GM-CSF protein either from circulating mononuclear cells or from alveolar macrophages. The expression of the GM-CSF mRNA was normal after the stimulation of lipopolysaccharide, whereas a point mutation at position 382 of the GM-CSF cDNA from "T" to "C" was revealed by cDNA sequencing, which caused a change in amino acid 117 of the protein from isoleucine to threonine. The beta c receptor expression on the cell surface was normal, and the beta c receptor mRNA expression and the sequence of the entire coding region of the beta c receptor were also normal. CONCLUSIONS: The decreased GM-CSF production is associated with the pathogenesis of human PAP. A point mutation of the GM-CSF cDNA may contribute to the decreased GM-CSF production in our adult PAP patient. The mutation of the beta c receptor in some of paediatric patients with PAP may not be a common problem in adult patients. PMID- 11930666 TI - Overexpression of bcl-2 protects hepatoma cell line HCC-9204 from ethanol-induced apoptosis. AB - OBJECTIVE: To investigate the effect of overexpression of bcl-2 on ethanol induced apoptosis of primary hepatocellular carcinoma (HCC) cells. METHODS: The retrovirus expression vector pDOR-SB containing human bcl-2 cDNA was introduced into a human HCC cell line HCC-9204 by liposome-mediated transfection. pDOR transfected and non-transfected HCC-9204 cells were used as controls. The expression of Bcl-2 protein by transfected HCC-9204 cells was detected by the immunohistochemical method. Then the cells were cloned with the limited dilution method continually until a monoclonal cell strain whose positive rate of Bcl-2 protein was 100% detected by flow cytometry was obtained. The killing rates of cells were detected by Methabenzthiazuron assay after the treatment of 6% ethanol for 6 h. The extent of apoptosis was analyzed by transferase-mediated dUTP nick end labeling (TUNEL) staining and flow cytometry. RESULTS: Most of the pDOR-SB transfected cells demonstrated Bcl-2 positive signals, while no signal was found in the controls. The positive rate of Bcl-2 protein detected by flow cytometry in the obtained monoclonal cell strain, which was named HCC-bcl2, was 100% after the cells had been cloned 3 times continually. The killing rate, TUNEL index and the scale of sub-G1 apoptotic peak in HCC-bcl2 cells were all significantly lower than those in the control cells. CONCLUSION: Overexpression of Bcl-2 protein suppresses ethanol-induced apoptosis of the HCC cell line HCC-9204. PMID- 11930668 TI - Localizing the focus of ischemic stroke with near infrared spectroscopy. AB - OBJECTIVE: To evaluate the changes in near infrared spectroscopy (NIRS) absorptive properties induced by cerebral ischemia. METHODS: A dual wavelength (760 nm and 850 nm) NIRS system measuring total hemoglobin concentration changes was constructed. Twenty Sprague-Dawley rats were used to set up middle cerebral artery occlusion (MCAO) model that had a stable ischemia focus on the cortex. We used NIRS to localize the ischemia focus that was confirmed with magnetic resonance imaging (MRI) and triphenyltetrazonlium chloride brain staining. The cortical ischemia area and the geometric configuration of the NIRS topograms were compared with those from MRI and the anatomical samples for the same rat. RESULTS: The reconstructed NIRS topograms showed that there was an optical density decreased area on the left cortex of the rats with MCAO model. The mean ischemia area as shown in NIRS images was 19.50 mm2 (19.50 +/- 0.35 mm2, n = 20). Anatomical samples showed that the mean ischemic area located in the NIRS measurement area was 18.46 mm2 (18.46 +/- 0.38 mm2, n = 20). For MRI, the mean ischemia area located in the NIRS measurement area was 20.71 mm2 (20.71 +/- 0.27 mm2, n = 20). There were no significant differences among NIRS, MRI and anatomical samples (F(2,57) = 2.47, P > 0.05) for defining the ischemia area. The results showed that there was a significant correlation among NIRS, MRI (r = 0.782, P < 0.05) and anatomical sample (r = 0.851, P < 0.05) for the same cortical ischemia area. Meanwhile, the location and geometric configuration of the ischemia focus shown by NIRS were identified by MRI and anatomical samples, though the NIRS images had worse spatial resolution. CONCLUSIONS: NIRS showed a good agreement with MRI and anatomical samples in the ischemic area and location determination of the infarction focus for the rat MCAO model. The study suggests that NIRS can non-invasively trace cortical hemodynamic changes induced by ischemia in real time. PMID- 11930667 TI - Acute effect of tetrandrine pulmonary targeting microspheres on hypoxic pulmonary hypertension in rats. AB - OBJECTIVE: To assess the effect of tetrandrine (Tet) pulmonary targeting microspheres on hypoxic pulmonary hypertension and evaluate its selective action on pulmonary circulation. METHODS: Twenty rats were exposed to hypoxic conditions for 3 weeks. Ten rats were used as normoxic controls. We administered Tet pulmonary targeting microspheres to 10 hypoxic rats and Tet aqueous solution to 10 hypoxic rats and the 10 control rats. Mean pulmonary arterial pressure (mPAP) was measured by a right cardiac catheterization, and mean systemic blood pressure (mSBP) was measured by left femoral catheterization. RESULTS: Rats exposed to hypoxia developed pulmonary hypertension. The decrease in mPAP in rats treated with Tet pulmonary targeting microspheres was significantly greater than that in rats receiving Tet aqueous solution (P < 0.05), and the effects were longer with Tet pulmonary targeting microspheres. Moreover, Tet pulmonary targeting microspheres, unlike Tet aqueous solution, did not decrease mSBP. CONCLUSION: Tet pulmonary targeting microspheres were more effective than Tet aqueous solution treating hypoxic pulmonary hypertension and acted selectively on the pulmonary circulation. PMID- 11930669 TI - D-AP5 blocks the increase of intracellular free Ca2+ induced by glutamate in isolated cochlear IHCs. AB - OBJECTIVE: To investigate the effect of D-AP5 (D-2-amino-5-phosphonopentanoate, a specific NMDA-antagonist) on the increase of intracellular free Ca2+ concentration ([Ca2+]i) induced by glutamate in isolated cochlear inner hair cells (IHCs), and to detect the autoreceptors of the IHC membrane. METHODS: When a laser scanning confocal microscope (LSCM) was used, the exogenous glutamate (Glu)-induced changes in [Ca2+]i of isolated IHCs and OHCs of guinea pig cochlea were observed with fluo-3, a fluorescent probe for [Ca2+]i. After D-AP5 or CNQX (6--cyano--7--nitroguinoxaline--2, 3--dione, a specific AMPA- antagonist) was administered, the exogenous glutamate (Glu)-induced changes in [Ca2+]i of isolated IHCs were recorded. RESULTS: In the presence of a low concentration Glu (3.85 mumol/L), there was an increase of [Ca2+]i in IHCs, whereas there was no change in OHCs. When 50 mumol/L of D-AP5 was administrated in advance, Glu did not induce a corresponding increase in [Ca2+]i in IHCs, and 50 mumol/L of CNQX did not completely block the increase of [Ca2+]i in IHCs. CONCLUSIONS: These results suggest that the autoreceptors existing in the IHC membrane are mainly of NMDA type, while there are relatively few AMPA receptors. Exogenous Glu is capable of increasing [Ca2+]i in IHCs by acting on the NMDA autoreceptor of IHCs in a positive feedback manner. PMID- 11930670 TI - Mechanisms of selective head cooling for resuscitating damaged neurons during post-ischemic reperfusion. AB - OBJECTIVE: To evaluate the efficacy and the mechanism of application of selective head cooling on neuronal morphological damage during postischemic reperfusion in a rabbit model. METHODS: 168 New Zealand rabbits were randomized into three groups. Group I [n = 24, (38 +/- 0.5) degrees C, non-ischemic control]; Group II [n = 72, (38 +/- 0.5) degrees C, normothermic reperfusion]; Group III [n = 72, (28 +/- 0.5) degrees C, selective head cooling, initiated at the beginning of reperfusion). Animals in three subgroups (n = 24, each) of Group II and Group III had reperfused lasting for 30, 180 and 360 min respectively. Using computerized image analysis technique on morphological changes of nucleus, the degree of neuronal damage in 12 regions were differentiated into type A (normal), type B (mild damaged), type C (severely damaged) and type D (necrotic). Fourteen biochemical parameters in brain tissues were measured. RESULTS: As compared with Group I, the counts of type A neuron decreased progressively, and those of type B, C and D increased significantly in Group II during reperfusion (P < 0.01). In Group II, vasoactive intestinal peptide, b-endorphine, prostacyclin, T3 and Na+, K(+)-ATPase were correlated with the changes of type A; b-endorphine and thromboxane with type B; glucose and vasopressin with type C; Na+, K(+)-ATPase, glutamic acid, T3 and vasoactive intestinal peptide with type D (P < 0.05). As compared with Group II, the counts of type A increased, and those of type C and D significantly decreased in Group III (P < 0.01). In Group III, Ca2+, Mg(2+) ATPase were correlated with the changes of type A, C and D (P < 0.01). CONCLUSION: Selective head cooling for sex hours during postischemic reperfusion does improve neuronal morphological outcomes in terms of morphological changes. PMID- 11930672 TI - [Paying attention to prevent and treats of secondary renal diseases]. PMID- 11930671 TI - DNA sequence analysis of the triose phosphate isomerase gene from isolates of Giardia lamblia. AB - OBJECTIVE: To confirm the genetic relation between Giardia lamblia (G. lamblia) isolates from different geographic regions of China and other countries. METHODS: Genomic DNA were extracted from the trophozoites or cysts of Giardia lamblia. The triose phosphate isomerase (tim) gene was amplified using polymerase chain reaction (PCR) technique. PCR products were digested with endonuclease and sequenced. The data of sequencing were analyzed with the DNAstar software and compared with that of the isolates acquired from GenBank. RESULTS: Of nine isolates of Giardia lamblia from China (C1, C2, CH2 and CH3), Cambodia (CAM), Australia (A1 and A2) and America (BP and CDC), respectively, 3 (A1, A2 and CAM) fit into Group 1 (WB), 2 (CH2 and CH3)) into Group 2, and 4 (C1, C2, BP and CDC) into Group 3 (GS). The results confirmed the genetic relatedness of G. lamblia isolates from all over the world. CONCLUSION: Genotyping isolates of G. Lamblia provides important information for establishing the phylogenetic relationship or for the epidemiological evaluation of the spreading of this organism. PMID- 11930673 TI - [Prevention of contrast-medium-induced acute renal failure]. PMID- 11930674 TI - [The expression of glucocorticoid receptor beta messenger RNA in peripheral white blood cells of hormone-resistant nephrotic syndrome patients]. AB - OBJECTIVE: To study the molecular mechanism contributing to hormone resistance of nephrotic syndrome. METHODS: The specific hormone binding sites of receptor was determined by radioligand binding assay. Messenger RNA of receptor was determined by quantitative reverse transcription-polymerase chain reaction. RESULTS: No significant differences of specific hormone binding sites of peripheral white blood cells were observed among hormone-sensitive patients, hormone-resistant patients and healthy volunteers. However, in hormone-resistant patients, GR alpha/GR beta mRNA ratio decreased significantly as compared with that of hormone sensitive patients and healthy volunteers. CONCLUSION: Results of the experiments indicated that the increased expression of GR beta, which leads to the decrease of the GR alpha/GR beta ratio, may be one of the mechanism responsible for hormone resistance in some nephrotic syndrome patients. PMID- 11930675 TI - [The association between polymorphism of endothelial nitric oxide synthase gene and diabetic nephropathy]. AB - OBJECTIVE: To investigate the association between a 894 G-->T mutation at exon 7 and a 27 base pair(bp) repeat polymorphism in intron 4 of the eNOS gene and diabetic nephropathy in Chinese. METHODS: A case control study of 228 Chinese subjects (including 143 type 2 diabetes mellitus with or without nephropathy and 85 normal controls) was performed. The number of 894 G-->T mutation allele and 27 bp repeat polymorhism alleles were determined by polymerasechain reaction restriction fragment lenth polymorphism(PCR-RFLP) method and PCR combined with 4% agarose electrophoresis. RESULTS: The frequency of the T allele and TG genotype at exon 7 and the a allele and ab genotype in intron 4 were significantly higher in diabetic nephropathy positive (DN+) group than in diabetic nephropathy negative (DN-) group and control subjects(P < 0.05). Diabetic patients with coexistence of the T and a alleles had a higher incidence of diabetic nephropathy(P < 0.05) than those with only one of the two alleles or without any of the two alleles. Multivariate logistic regression analysis showed that blood glucose, GHbA1c, SBP, TC, 894 G-->T mutation at exon 7 and a 27 base pair(bp) repeat polymorphism in intron 4 of the eNOS gene are independent risk factors for diabetic nephropathy. CONCLUSIONS: The T allele at exon 7 and a allele in intron 4 are related to diabetic nephropathy in Chinese patients with type with 2 diabetes mellitus. The incidence of diabetic nephropathy is higher in patients who have both T and a alleles than patients who have either T or a allele alone. PMID- 11930676 TI - [A prospective study on the effects of low-osmolar and nonionic contrast media on renal functions]. AB - OBJECTIVE: To investigate the effects of contrast medium, especially large dose of Iopromide, on renal functions. METHODS: 100 hospitalized patients who were going to have cardiac or renal angiography were selected. 96 patients had normal renal functions and 4 had slightly abnormal renal functions. Scr, Ccr, urine gamma-GT and urine glucose were monitored on the first, third and fifth day after angiography. RESULTS: All the 100 patients were administered with Iopromide. Of the 96 patients with normal renal functions, all had elevated urine gamma-GT on the first day after giving Iopromide; the level returned to normal on the third day. There was no significant change of Scr and Ccr after angiography. Of the 4 patients with abnormal renal functions, all had significant change of Scr and Ccr after using Iopromide and the levels of both returned to pre-angiography level on the fifth day. CONCLUSIONS: In patients with normal renal functions, Iopromide may cause transient elevation of urine gamma-GT level and positive urine glucose. In patients with slightly abnormal renal functions, Iopromide may cause significant decrease of Ccr and increase of Scr. However, with proper treatment, these indexes may return to pre-angiography level on the fifth day after angiography. PMID- 11930677 TI - [Comparison of mortality rates between peritoneal dialysis and hemodialysis in diabetic nephropathy patients]. AB - OBJECTIVE: To evaluate and compare the different survival rates and clinical outcome of continuous ambulatory peritoneal dialysis (PD) and hemodialysis (HD) treatment in diabetic patients. METHODS: 62 diabetic patients suffering from end stage renal complication were treated with PD or HD. The age, sex and residual renal function of the two groups were matching. Their survival rates and levels of blood lipid, blood sugar, serum albumin and hemoglobin were observed. RESULTS: The mortality rates of HD and PD were not significantly different in the first two years. But mortality of PD increased rapidly after two years of treatment. The levels of blood triglycerides, low density lipoprotein cholesterol and fasting sugar in PD patients were higher than that in HD patients, while the concentration of serum albumin was lower than HD patients significantly. CONCLUSIONS: In diabetic patients with renal failure PD as a choice to replace the kidney function was not superior to HD, especially after two years of PD treatment. A close watch is necessary to detect the change of parameter levels mentioned above and if is better to shift to HD. A strict long-term and extensive survey is needed to compare these two methods further. PMID- 11930678 TI - [Diagnostic significance of measurement of the receptor for urokinase-type plasminogen activator on granulocytes and in plasma from patients with paroxysmal nocturnal hemoglobinuria]. AB - OBJECTIVE: To investigate the expression levels of the receptor for urokinase type plasminogen activator (uPAR) on granulocytes and the soluble uPAR (suPAR) level in plasma from patients with paroxysmal nocturnal hemoglobinuria (PNH) and its clinical application in the diagnosis of this disorder. METHODS: Expression of uPAR, CD55 and CD59 was measured on granulocytes by using flow cytometry (FCM) and the level of suPAR in plasma by using immunoradiometric assay (IRMA) in 20 patients with PNH, 59 anemic patients with diagnoses other than PNH (18 with autoimmune hemolytic anemia, 6 with other hemolytic anemias, 26 with aplastic anemia and 9 with iron deficiency anemia) and 21 healthy individuals. RESULTS: In PNH patients, both the mean fluorescence intensity and percentage of positive fluorescence-activated granulocytes of uPAR, CD55 and CD59 were remarkably decreased, while in anemic patients with diagnoses other than PNH the results were not different when compared with those of the healthy individuals. The expression level of uPAR was reduced to the same extent as that of CD55 and CD59 on the PNH-affected granulocytes. In addition, some peak shape abnormalities (double peaks and/or peak tailing) in the histogram of fluorescence intensity were also found in PNH patients. The suPAR concentration of PNH plasma was (4.04 +/- 2.47) micrograms/L, being higher than that of the healthy individuals (1.73 +/- 0.96) micrograms/L (P < 0.01). And the percentage of positive fluorescence activated granulocytes was inversely associated with the plasma suPAR level in PNH patients (r = -0.79, P < 0.01). CONCLUSION: It is suggested that measurement of expression level of uPAR on granulocytes with FCM and suPAR in plasma with IRMA would be alteanative specific techniques for the diagnosis of PNH. PMID- 11930679 TI - [The relationship among plasma soluble vascular cell adhesion molecule-1 level, airway responsiveness and blood eosinophil cationic protein in asthmatics with remission at least 3 years]. AB - OBJECTIVE: To study the mechanism of asthma remission. The plasma soluble vascular cell adhesion molecule-1 (sVCAM-1), airway responsiveness (PC20) and other relative factors were measured in asthmatics with remission at least 3 years and compared with healthy controls. METHODS: Group I (asthmatics with remission at puberty): 20 cases including 11 males and 9 females, mean age of 21 years. All of the patients were definitely diagnosed as asthmatics in childhood and got natural remission for at least 3 years(with median duration of 6 years). Group II (normal controls): 21 healthy college students including 10 males and 11 females, mean age of 22.5 years, had no history of asthma or other allergic diseases. The intravenous blood was taken from all subjects with EDTA as anticoagulant for study. The airway responsiveness were measured by methacholine bronchial provocation test(PC20). The sVCAM-1 was measured with ELISA, according to the manual of the kit. Methacholine was atomized by Wright atomizer [Devilbiss AP-50, USA, driven by air compressor, with a constant atomizing volume of (0.13 +/- 0.02) ml/min, diameter of atomizes particles 1.3-3.0 microns]. Its concentration was redoubled from 0.03 to 128 g/L until a certain dose as PC20 value triggering forced expiratory volume in one second (FEV1) decreased by 20% reached. The eosinophil cationic protein (ECP) levels were measured with enzyme linked immuno-CAP system. Blood eosinophils(> 330/mm3 as being above normal) and basophils (> 33/mm3 as abnormally increased) were counted using Acian blue dyeing. RESULTS: (1) The mean value of sVCAM-1 of group I (63.7 +/- 31.7) micrograms/L was significantly higher than that of group II (41.9 +/- 11.7) micrograms/L. (2) The percentage of the cases with increased eosinophils in group I was 15.0% (3/20), had no significant difference compared with that in group II (4.8%, 1/21). The percentage of the cases with increased basophils of group I was 45.0%(9/20), significantly higher than that of group II, 4.8%(1/21), P < 0.01. (3) The blood ECP level of group I was (2.7 +/- 1.3) micrograms/L, simillar to that of group II (2.18 +/- 0.5) micrograms/L. (4) The PC20 value of group I (M = 2.6 g/L) was significantly lower than that of group II (M = 128 g/L). (5) The basophil count negatively correlated with the PC20(r = -0.66), however, the sVCAM 1 did not have significant correlation with the basophil count or with the PC20. CONCLUSION: The quantity and activity of eosinophils in asthmatics with remission at least 3 years are droped to the normal level, but the expression of VCAM-1, quantity of basophils were still higher than normal. The airway responsiveness had negative correlation with the basophil count. PMID- 11930680 TI - [Clinical characteristics of primary biliary cirrhosis with Sjogren's syndrome of 20 cases]. AB - OBJECTIVE: Primary biliary cirrhosis (PBC) is a chronic non-suppurative destructive cholangitis of unknown etiology. It has been reported that patients with PBC suffer from xerostomia and keratoconjunctivitis at high prevalence and PBC has been shown to be associated with Sjogren's syndrome. This study investigated the relationship between PBC and Sjogren's syndrome. METHOD: 20 cases diagnosed as PBC consented to the evaluation for Sjogren's syndrome which included clinical and serological features, Schirmer's test, measurement of parotid flow rate and labial minor salivary gland biopsy. RESULTS: The main manifestations of PBC are jaundice, pruritus, hepatosplenomegaly. M2 subtype of antimitochondrial antibody is of great specificity for PBC. Many PBC patients are suffering from xerostomia and keratoconjunctivitis sicca but do not have typical serological and pathological presentations of primary Sjogren's syndrome. Corticosteroids and immunosuppressive agents have disappointing effects in the treatment. Ursodeoxycholic acid is effective to certain extent. CONCLUSION: It was indicated that Sjogren's syndrome associated with PBC appears to be a secondary form differing from primary Sjogren's syndrome. PMID- 11930681 TI - [A study for the relationship between serum calcitonin gene-related peptide and coronary arteriopathy]. AB - OBJECTIVE: To investigate the relationship between serum calcitonin gene-related peptide (CGRP) and coronary angiopathy. METHODS: In a cross-sectional study, serum CGRP levels of 119 patients with coronary heart disease (CHD), 78 with non CHD and 39 healthy subjects were measured with radioassay method. The information on conventional risk factors were collected by interviews. RESULTS: The development of coronary angiopathy was correlated with smoking, diabetes and aging. The mean serum CGRP level was significantly lower in CHD than in NCHD patients [(35.5 +/- 48.8) mumol/L vs (63.8 +/- 47.3) mumol/L, P < 0.01). There was no difference between the mean serum CGRP level in NCHD patients and healthy controls. By multivariate logistic regression, the odds ratios (OR) of aging, smoking and diabetes were all > or = 1 (P < 0.01), that means they are independent risk factors for the development of coronary angiopathy. The OR of CGRP was < or = 1, indicating that it was an independent protective factor. CONCLUSIONS: It is shown that aging, smoking and diabetes are all independent risk factors for the development of angiopathy, while CGRP is an independent protective factor for the development of coronary angiopathy. PMID- 11930682 TI - [Clinical and pathological characteristics of heroin spongiform leukoencephalopathy in China]. AB - OBJECTIVE: To observe the clinical, image and pathological characteristics of Heroin Spongiform Leukoencephalopathy(HSLE). METHODS: Clinical, CT and MRI analysis of 28 cases, brain autopsies of 2 cases and brain biopsies of 8 cases with HE, Loyez and Congo Red staining and observation through electron microscope. RESULTS: The clinic, image and pathologic characteristics of HSLE: (1) the history of inhalation of heated heroin vapor; (2) among 28 cases with HSLE, recurrence of HSLE was found in 17 cases during the abstinence, in 8 cases 1-2 month after abstinence, in 2 cases 4 month after abstinence and in 1 case after inhaling large amount of heroin; (3) the majority of cases were described with the acute onset characterized by cerebellar signs; (4) pyramidal tract lesion was frequently involved, but sensory system usually remained normal; (5) CSF test was normal; (6) CT and MRI revealed extensive symmetric white matter lesions in cerebra and cerebellum; (7) spongiform vacuoles degeneration of white matter was the main morphological change. CONCLUSIONS: Spongiform leukoencephalopathy may be considered if a patient who had a history of inhaling heated heroin vapor showed acute cerebellar signs. Spongiform vacuoles degeneration of cerebral white matter was the main pathological change. PMID- 11930683 TI - [Amphotericin B for treatment of fungal infections in 40 patients with malignant hematologic diseases]. AB - OBJECTIVE: To observe the therapeutic effect and side effects of amphotericin B for fungal infections in patients with malignant hematologic diseases. METHODS: 40 patients (male 27, female 13; average age 35.5 years) with malignant hematologic diseases were given amphotericin B, 5-50 mg/d per day for 5-85 days (average time 21 days). RESULTS: The clinical efficacy rate of amphotericin B was 52.5%, and the fungal elimination rate was 56.2%. Among the side effects, rigor and fever were present in 2.5% of the patients. Hypokalaemia was found in 12.5%, hepatotoxicity in 15.0% and nephrotoxicity in 15.0%. CONCLUSION: As amphotericin B has a broad anti-fungal spectrum and relatively good efficacy, it is still a high-efficiency drug in treatment of systematical fungal infections. However, the use of drug is limited because of its many side effects. Our study indicates that if it is used properly and hepatic and renal function tests are carried out regularly, amphotericin B is a relatively safe and effective drug. PMID- 11930684 TI - [A clinical study of haploidentical transplantation using granulocyte colony stimulating factor stimulating donor bone marrow]. AB - OBJECTIVE: To explore the effects of reducing the incidence of severe acute graft versus-host disease (GVHD) and improving the disease free survival(DFS) in haploidentical donor transplantation by granlocyte colony-stimulating factor (G CSF) administration to donor before harvesting and a number of immunosuppresants added to host. METHODS: Thirteen patients with leukemia received allo-bone marrow transplantation (BMT) from two or three HLA loci mismatched related donor (haploidentical group). The clinical outcomes of the bone marrow transplantion were compared with thase of 13 consecutive HLA identical sibling transplantion (identical group). In haploidentical donor BMT, the donors of patients were given G-CSF (Lenograstim Chugai) 250 micrograms/day for seven doses prior to marrow harvest. CSA, MTX, ATG and mycophenolate mofetil (MMF) were combined for GVHD prophylaxis. ATG 5 mg/kg/day was infused for 4 days before transplantation and MMF was adminisered from 7th day after. RESULTS: All the patients were engrafted. The median number of CD34+ cells in graft was 6.1 x 10(6)/kg in haploidentical group and 2.5 x 10(6)/kg in identical group (P < 0.01). The median number of CD3+ cells was 50.5 x 10(6)/kg and 47.0 x 10(6)/kg respectively (P > 0.05). All patients had 100% donors hematopoietic cells after transplantation by cytogenetic evidence analysis. Five of the thirteen patients (38.5%) in haploidentical group and three of the thirteen patients(23.1%) in identical group experienced II-IV acute GVHD (P > 0.05). The probability of chronic GVHD was 87.5% in haploidentical group and 67.5% in identical group (P > 0.05), However none in both groups developed extensive cGVHD. The median follow-up duration was 453 days (range 180-690 days) for haploidentical group and 510 days (range 220-810 days) for identical group. In haploidentical group, five patients died from transplant related mortality (3 GVHD, 2 infection), none relapsed and eight patients(61.5%) survive in disease free situation. In identical group, two patients died from transplant related mortality (1 GVHD, 1 infection), two patients died from relapse and nine patients (69.2%) survive in disease free situation. DFS in haploidentical group and in identical group was similar(P > 0.05). CONCLUSION: The transplants from haploidentical donor used in this study is 3 effective and feasible in preventing acute severe GVHD and improving DFS. PMID- 11930685 TI - [Study on ovarian hyperstimulation syndrome]. PMID- 11930686 TI - [Association between renin activity, angiotensin II and ovarian hyperstimulation syndrome]. AB - OBJECTIVE: To investigate the association between renin-like activity (RA) and angiotensin II (A II) and the ovarian hyperstimulation syndrome (OHSS). METHODS: Blood samples were taken from 42 patients undergoing in vitro fertilization (IVF). According to the ovarian stimulating response, the patients were divided into 4 groups: group I, low responders, 7 cases; group II, moderate responders, 8 cases; group III a high responders without using albumin, 7 cases, group III b, high responders with albumin, 10 cases; group IV, severe and moderate OHSS, 10 cases. 5 patients for intrauterin insemination with natural cycle as control. Follicular fluid (FF) was collected at oocyte retrieval from 28 IVF patients (including 7 OHSS patients). Fluid from ascites and hydrothorax was obtained from 3 OHSS patients, and peritoneal fluid obtained from 5 infertility patients by laparoscopy and ascitic fluid obtained from 6 ovarian carcinoma patients served as control. RA and A II levels were measured by radioimmunoassay. RESULTS: Plasma RA and A II levels at mid-luteal phase from OHSS patients [(19.9 +/- 19.0) micrograms.L-1.h-1 and (397.0 +/- 378.2) ng/L] were significantly higher than those from the other patients (P < 0.01-0.05), also the levels at mid-luteal phase from IVF patients were significantly higher than those at late-follicular phase [(4.1 +/- 2.9)-(4.9 +/- 3.2) micrograms.L-1.h-1 Vs (1.5 +/- 0.9)-(1.9 +/- 1.0) micrograms.L-1.h-1, (85.5 +/- 49.5)-(109.4 +/- 46.0) ng/L Vs (33.6 +/- 15.9) (37.0 +/- 17.1) ng/L, P < 0.01-0.05]. RA and A II levels in FF were significant higher than those in plasma (P < 0.01-0.05). A II in ascitic fluid in OHSS patients was 4 to 8 times higher than that in plasma, while RA in ascitic fluid was 1.5 to 3 times higher than that in plasma, but the levels of A II and RA in controls were very low. The plasma RA and A II levels dropped very significantly along with clinical improvement (P < 0.01). CONCLUSION: These findings suggest that RA and A II are associated with the pathogenesis of capillary leakage in OHSS. PMID- 11930687 TI - [Function of vascular endothelial growth factor in ovarian hyperstimulation syndrome]. AB - OBJECTIVE: To evaluate the function of vascular endothelial growth factor (VEGF) and other clinical indexes to forcast ovarian hyperstimulation syndrome (OHSS). METHOD: Collecting the serum and follicular fluid (FF) of 42 cases in the in vitro fertilization and embryo transfer (IVF-ET) cycles and then analysis the relationship of the clinical and detected indexes between the OHSS (10 cases) and the non-OHSS (32 cases). RESULTS: The concentration of VEGF in FF, E2 on the day of human chorionic gonadotropin (hCG) injection, basal luteinizing hormone (LH) and the number of ovum retried much higher in the OHSS than in the non-OHSS. CONCLUSIONS: The concentration of VEGF in FF of OHSS cases is higher than that of controls, supporting the role of VEGF as a mediator of OHSS. Therefore VEGF in FF is a forcast index of OHSS. PMID- 11930688 TI - [Preliminary study on the role of vascular endothelial growth factor in pathogenesis of ovarian hyperstimulation syndrome]. AB - OBJECTIVE: To explore the role of vascular endothelial growth factor (VEGF) in the pathogenesis of ovarian hyperstimulation syndrome (OHSS). METHODS: Fourteen moderate or severe OHSS patients and 13 non-OHSS cases were enrolled prospectively from in vitro fertilization and embryo transfer (IVF-ET) or intracytoplasmic sperm injection program. The VEGF levels of follicular fluid (FF) on the day of oocytes pick up (DOPU) and of sera on the day of human chorionic gonadotrophin (hCG) injection (DhCGI) and embryo transfer (DET) were measured by enzyme-labeled immunosorbent assay. Expression of VEGF protein and mRNA of luteinized granulosa cells obtained during OPU were detected by immunohistochemical and reverse transcription polymerase chain reaction respectively. RESULTS: The VEGF levels of FF on the DOPU and of sera on DhCGI were not significantly different between the OHSS and control groups [1257.2 +/- 648.0) ng/L Vs (1745.1 +/- 802.4) ng/L and (250.1 +/- 109.5) ng/L Vs (196.7 +/- 81.7) ng/L, respectively, P > 0.05]. However, both serum VEGF levels on DET and its increments from DhCGI to DET are significantly greater in the OHSS group as compared with the controls, so was the expression of VEGF mRNA levels of luteinized granulosa cells on DOPU. CONCLUSION: VEGF may play a role in pathogenesis of OHSS. PMID- 11930689 TI - [Expression of leukemia inhibitory factor in the decidua of normal early pregnancy, threatened abortion and inevitable abortion]. AB - OBJECTIVE: To investigate the expression of leukemia inhibitory factor (LIF) in the decidua of normal early pregnancy, threatened abortion and inevitable abortion. METHOD: We examined LIF gene expression in the above-mentioned decidua by a quantitative reverse transcription-polymerase chain reaction (RT-PCR) method, and also examined the serum pregesterone, human chorionic gonadotrapin (hCG) by radioimmunoassay in all cases. RESULTS: (1) Serum levels of pregesterone and hCG are: (91.5 +/- 27.2) nmol/L, (69.9 +/- 14.9) kU/L in normal early pregnancy; (88.4 +/- 24.7) nmol/L, (57.6 +/- 11.2) kU/L in threatened abortion respectively. There was no difference in the levels of pregesterone and hCG between the two groups (P > 0.05). While serum pregesterone, hCG levels in inevitable group were (33.1 +/- 19.6) nmol/L, (10.3 +/- 3.2) kU/L respectively. Compared with normal early pregnancy and threatened abortion group, the levels of serum pregesterone and hCG reduced significantly (P < 0.05). (2) The expression of LIF in three groups: There was no statistically significant difference in the levels of LIF expression between the normal early pregnancy group (2.10 +/- 0.32) and threatened abortion (1.92 +/- 0.20) groups, while the levels of LIF expression in inevitable abortion group (0.7 +/- 0.06) was lower than those in normal early pregnancy group and threatened abortion group (P < 0.05, respectively). CONCLUSION: The reduction of LIFmRNA expression in the decidua of early pregnancy may decrease the serum pregesterone and hCG levels and cause inevitable abortion. PMID- 11930690 TI - [The changes of serum levels of interleukin-6 and interleukin-8 in pregnant women with systemic lupus erythematosus]. AB - OBJECTIVE: To assess the changes of serum levels of interleukin-6(IL-6) and interleukin-8 (IL-8) in pregnant women with stationary phase systemic lupus erythematosus (SLE). METHODS: Enzyme-linked immunosorbent assay (ELISA) was used to measure serum IL-6 and IL-8 from 14 pregnant women with stationary phase SLE (SLE group), 12 normal pregnant women (normal pregnant group) and 12 normal non pregnant women (group normal non-pregnant). The later two groups are set as controls. RESULTS: The IL-6, IL-8 levels in group SLE patients are (20.31 +/- 5.70) ng/L and (48.80 +/- 9.17) ng/L, significantly higher than those in normal pregnant group [IL-6, IL-8 levels are (8.40 +/- 2.49) ng/L and (21.15 +/- 5.21) ng/L, respectively, P < 0.01)] and normal non-pregnant group [IL-6, IL-8 levels are (6.14 +/- 0.86) ng/L and (17.71 +/- 4.43) ng/L, respectively, P < 0.01]. However, there's no significant difference between the normal pregnant group and normal non-pregnant group (P > 0.05). CONCLUSION: Serum levels of IL-6, IL-8 may be helpful to monitor the progress of SLE during pregnancy. PMID- 11930691 TI - [Study on endothelial cell injury in women with pregnancy induced hypertension]. AB - OBJECTIVE: To investigate the mechanisms of endothelial cell injury in women with pregnancy induced hypertension (PIH). METHODS: Human umbilical endothelial cells were cultured in the serum from 10 women with severe PIH and 10 normal pregnant women, and co-cultured with neutrophils or not for 48 hrs. The morphology and cell cycles were studied. RESULTS: Without neutrophils, no apoptotic cells were observed, while neutrophils were added, apoptotic cells were obviously detected. The percentage of apoptotic cells in PIH was more than that in non-PIH. The percentage of cells in G2/M phase decreased in all the groups, but there were no significant differences among them. CONCLUSIONS: Endothelial cells were injured indirectly in PIH patients. Activated neutrophils induced apoptosis might be one of the answers. PMID- 11930692 TI - [Percutaneous testicular sperm aspiration and intracytoplasmic sperm injection in the treatment of severe male infertility]. AB - OBJECTIVE: To investigate the efficacy of intracytoplasmic sperm injection (ICSI) combined with percutaneous testicular sperm aspiration (PTSA) in the treatment of severe male infertility. METHODS: From October 1998 to December 2000, 162 couples were enrolled, and ICSI and PTSA were adopted. Ovarian stimulation was achieved by the short protocol. All metaphase II (M II) oocytes were selected for ICSI. RESULTS: 1,517 M II oocytes were injected in 185 cycles, 990 fertilized (65.3%), and 152 embryo implanted. 54 couples achieved clinical pregnancy (35.5%). CONCLUSION: PTSA combined with ICSI is a rapid, convenient, painless and effective approach for the treatment of severe male infertility. PMID- 11930693 TI - [Study on the reversion of drug resistance in human cervical cancer cell lines]. AB - OBJECTIVE: To determine the resistance reversion of mitomycin (MMC) by 3'-Keto bmt1-val2-cyclosporin (SDZ PSC 833) in human cervical cancer in vitro and in vivo. METHEDS: A xenografted mitomycin resistant mice model of cervical cancer was devolped. The reversion of mitomycin resistance by SDZ PSC 833 (1 or 3 mg/L) was detected from human cervical cancer cell (Hela) and its resistant subline Hela/MMC in vitro and in vivo. Studies in vitro include drug resistance reversion experiment and the changes of morphology. Studies in vivo including tumor volume and tumor related histopathological changes in the autopsied specimen were evaluated by comparing random sections of each group. RESULTS: Nontoxic doses of SDZ PSC 833 could result in almost partial reversion of MMC-resistance of Hela/MMC. In vivo studies also showed SDZ PSC 833 augmented the growth inhibitory effect of mitomycin on Hela/MMC xenografted in nude mice. CONCLUSION: SDZ PSC 833 can overcome mitomycin resistance of Hela/MMC in vitro and in vivo, so SDZ PSC 833 will be a better candidate clinically for reversing multidrug resistance. PMID- 11930694 TI - [Restaging laparotomy of presumed early ovarian cancer]. AB - OBJECTIVE: To assess the value and complications of restaging laparotomy in women with presumed early ovarian cancer who have undergone inadequate initial staging procedures. METHODS: Between 1986 and 1996, 42 patients underwent restaging laparotomy in Cancer Hospital, Peking Union Medical College after receiving inadequate initial surgical procedure for presumed early ovarian cancer. Presumed stages from initial surgery include stage I a in 28 cases; I b in 1 case; I c in 12 cases; II a in 1 case. Histological distributions were as follows: epithelial cancer in 26 cases; malignant germ cell tumor in 9 cases; granulose cell tumor in 7 cases. RESULTS: Twelve patients (28.6%) had disease upstaged and eight (19.0%) had stage III disease confirmed by restaging laparotomy. Positive findings at restaging laparotomy were mainly in omentum (16.7%) and pericolic gutters (33.3%). Peritoneal cytology was positive in 25.0 percent of patients. Three and five year survival rates for 30 patients with negative findings were 96.7% and 86.7%, respectively. Complications of restaging laparotomy included hemorrhage blood loss (50-1,100 ml, average 280 ml) and lymphocyst (7.1%). CONCLUSIONS: Patients with presumed early ovarian cancer who had undergone inadequate staging procedures should undergo restaging laparotomy. Patients with negative findings have an excellent prognosis. Complications of restaging laparotomy were minor. PMID- 11930696 TI - [Follow the guideline of pulmonary thromboembolism]. PMID- 11930695 TI - [Clinical study on women with amenorrhea after levonorgestrel intrauterine system]. AB - OBJECTIVE: To study the 5 years contraceptive efficacy of levonorgestrel intrauterine system (LNG-IUS) and the ovarian functions in amenorrhea women. METHODS: LNG-IUS and TCu 380A IUD were randomly inserted in 2 groups of women, 50 each. Women were followed annually for 5 years. Serum follicle stimulating hormone (FSH), luteinizing hormone (LH), prolactin (PRL), estradiol (E2), and T levels in 14 women with amenorrhea, and endometrial biopsies were obtained in 9 cases. RESULTS: By the end of 5 years, there were 2 pregnancies in TCu 380A group, but none in LNG-IUS group; and 24 amenorrhea cases in LNG-IUS. The 5 year cumulative continuation rates were 24% for LNG-IUS and 78% for TCu 380A (P < 0.001). In the 14 women with amenorrhea, the average serum concentrations were not significantly different from that of the normal controls. Histological study showed endometrial atrophy or a few secretory glands. CONCLUSIONS: LNG-IUS is a highly effective and safe contraceptive measure. Oligomenorrhea and amenorrhea caused by the local effects of LNG were reversible and did no harm to women's health. However, comprehensive consultation provided to women before insertion is of utmost importance for continuation use. PMID- 11930697 TI - [Trends of underline diseases of pulmonary embolism from Peking Union Medical College Hospital]. AB - OBJECTIVE: To highlight the understanding of pulmonary embolism(PE), and analyze the trends of underline diseases of PE from Peking Union Medical College(PUMC) Hospital in the last half century. METHODS: 239 cases of PE were reviewed retrospectively from 1950 to 2000 in PUMC Hospital. RESULTS: In the first and second periods (1950-1982, 1983-1990), about 3 cases of PE were diagnosed annually, but in the third (1991-1997) and fourth periods (1998-2000), 8 cases and 20.6 cases of PE were found annually. The incidence of underline diseases of PE, such as deep venous thrombosis (DVT), cardiac disease, malignancy and connective tissue disease varied in different periods. The morbidity of DVT with PE was dramatically enhanced recently. CONCLUSION: There is an increasing trend of incidence of PE in the last half century. DVT events become the most common underline disease or risk factor of PE. PMID- 11930698 TI - [The significance of thrombosis after experimental pulmonary thromboembolism]. AB - OBJECTIVE: To study the significance of thrombosis after experimental pulmonary thromboembolism (PTE). METHODS: Acute PTE models of rabbits were established with injection of autologous blood clots (0.04 g/kg) stabilized in a temperature controlled (70 degrees C) of distilled water for 10 minutes through the femoral vein, then the regulation of thrombosis was explored at dissection and upon microscopic examination after PTE. Moreover, the coagulability of blood and the plasma level of thromboxane A2(TXA2) and endothelin (ET) were examined. RESULTS: Thrombotic propensity was found at 1 h, and fresh thrombosis started to form at 24 h following clots infusion. Emboli were completely or partly dissolved at 5 d and organized at 10 and 14 d after clots infused. Prothrombin time was significantly lower [(7.15 +/- 0.06)s], and fibrinogen was higher [(5.86 +/- 1.50) g/L] at 24 h post-clots, compared with pre-clots [(7.34 +/- 0.19)s, (3.37 +/- 1.02) g/L] (P < 0.05). Venous plasma level of TXA2 began to increase at 5 min [(2.5 +/- 0.7) micrograms/L] and continued to rise to its maximum at 15 min [(2.5 +/- 0.6) micrograms/L], then declined at 60 min after clots infusion. The level of ET in both arterial and venous blood increased at 5 d post-clots [(0.84 +/- 0.15) micrograms/L and (0.23 +/- 0.05) micrograms/L] separately, while most of emboli resolved. CONCLUSIONS: There is thrombus formation after autologous-blood clots-induced PTE. Furthermore, thrombus formation, fibrinolysis and organization may always interact on each other consistently, and control the pathogenesis of PTE. Abnormalities of ET metabolism occur after PTE and the major mediator of TXA2 plays an important role in the early phase of PTE. PMID- 11930699 TI - [The prognostic value of serum neuron specific enolase detection in small cell lung cancer]. AB - OBJECTIVE: To evaluate the prognostic value of NSE detection in small cell lung cancer. METHODS: 144 unresectable small cell lung cancer patients were eligible in the retrospective study. NSE was analyzed before chemotherapy and 21 days after chemotherapy using ELISA method. RESULTS: For D1-NSE, 98 (68.1%) were NSE positive and 46(31.9%) were NSE negative; for D21-NSE, 57(39.6%) were NSE positive and 87 (60.4%) were NSE negative. In univariate analysis, both D1-NSE and D21-NSE have prognostic significance. But in multivariate analysis only D21 NSE is an independent prognostic factor. CONCLUSION: D21-NSE in small cell lung cancer is an independent prognostic factor and may be proposed for use in the clinic and research. PMID- 11930700 TI - [Synthesis and release of pulmonary tissue adrenomedullin on hypoxic pulmonary hypertension in rats and its significance]. AB - OBJECTIVE: To investigate the synthesis and release of adrenomedullin (AM) of lung tissue in rats on hypoxic pulmonary hypertension (HPH) and study its mechanism and significance in the pathogenesis process of HPH. METHODS: 54 Wistar rats were divided into hypoxia 10 d (n = 12), 20 d (n = 12), 30 d (n = 12) groups and control group (n = 18). The rats of hypoxia groups were exposed to chronic hypobaric hypoxia environment at high 5 km to establish HPH models. The expression of pulmonary cell AM protein, the content of plasma AM and bronchial alveolar lavage fluid (BALF) AM were determined by optical microscopy, immunohistochemistry and radio-immunoassay. RESULTS: AM was widely distributed in lung tissue. There was positive of AM in intraplasma of vascular endothelial cells (EC), vascular and bronchial smooth muscular cells (SMC), bronchial epithelial cells, macrophages (M phi) and type II alveolar epithelial cells in every groups. After hypoxia, the expression of AM in above cells, especially EC, SMC and M phi, increased more than control group. Plasma AM contents of hypoxia groups were higher than control group(P < 0.01). 20 d after hypoxia, the AM content was twice as control group. At 10 d to 20 d after hypoxia, BALF AM content was increased more than control group (P < 0.01). But at 30 d the content was decreased near to normal. CONCLUSIONS: Hypoxia is effective in promoting AM synthesis and release in pulmonary tissue and suggestes that AM as a peptide plays an important regulating role on pulmonary circulation, exchange of air and pulmonary vascular structural remodeling in the pathophysiological process of HPH. PMID- 11930701 TI - [Low flow technique to perform static pressure-volume curve during mechanical ventilation]. AB - OBJECTIVE: To describe a fast, simple method to acquire static pressure-volume (P V) curve of the respiratory system and to compare it with airway occlusion technique. METHODS: ARDS sheep model was induced by intravenously infusing 3-6 micrograms/kg LPS, static P-V curves were determined by low flow technique and airway occlusion technique. Static compliance (Cst), lower inflection point (Pinfd and Pinfb) and airway pressure were compared between the two methods. RESULTS: Pinf were (8.91 +/- 0.82) cm H2O and (8.59 +/- 0.78) cm H2O determined by low flow technique and airway occlusion technique respectively, the correlation between which was good (r = 0.93, P < 0.05). Cst determined by low flow technique was similar to that determined by airway occlusion technique [(19 +/- 7) L/cm H2O vs (20 +/- 7) L/cm H2O, P > 0.05]. The time needed to acquire a static P-V curve by low flow technique was 3-4 minutes versus 30-35 minutes by airway occlusion technique. CONCLUSION: Accuracy and reliability of low flow technique are sufficient for clinical use, it is much less time consuming than the airway occlusion technique. PMID- 11930702 TI - [Changes of L-selectin expression on polymorphonuclear leukocytes and its role during polymorphonuclear leukocytes sequestration in lungs of rats with acute lung injury]. AB - OBJECTIVE: To explore the changes of L-selectin protein expression on peripheral blood polymorphonuclear leukocytes (PMN) and their role during PMN sequestration in the lungs of rats with acute lung injury (ALI). METHODS: The model of ALI in rat was established by intravenous injection of E. Coli endotoxin (ET). The protein expression of L-selectin on peripheral blood PMNs was measured by immunofluorescence and flow cytometry. Myeloperoxidase (MPO) enzyme analysis and histology were employed to estimate the quantity of PMN sequestration in lungs of rats during ALI. RESULTS: (1) The protein expression of L-selectin on PMN after injection of ET (7.8 +/- 1.6) was all significantly lower than that in control group (10.5 +/- 2.1, P < 0.05). (2) MPO activity increased significantly after injection of ET in comparison with the control group [0.069 +/- 0.011) U/mg lung tissue, P < 0.01]. The difference was not significant between the fucodin treated 5 min group [(0.391 +/- 0.071) U/mg lung tissue] and ET-treated 5 min group [(0.359 +/- 0.074) U/mg lung tissue]. But MPO activity was significantly lower in fucodin-treated 15 min group [(0.396 +/- 0.061) U/mg lung tissue] than that in ET treated group at same time point [(0.490 +/- 0.069) U/mg lung tissue, P < 0.05]. CONCLUSIONS: (1) In physiological state, L-selectin are consistently expressed on the surface of PMN membrane. The protein expression of L-selectin on PMNs reduces rapidly after injection of ET and is the lowest at 15 min, then gradually increases. (2) The early stage of PMN sequestration in lungs is not dependent on L-selectin, but L-selectin is still important for maintaining a continual PMN sequestration in lungs of rats with ALI. PMID- 11930703 TI - [Construction of Ag85B DNA vaccine for Mycobacterium tuberculosis and its immunogenicities]. AB - OBJECTIVE: To construct pcDNA3-Ag85B recombinant and evaluate its immunogenicity and types of immune response induced by recombinant pcDNA3-Ag85B. METHODS: Ag85B gene of Mycobacterium tuberculosis was inserted into pcDNA3 to construct DNA vaccine pcDNA3-Ag85B. BALB/c mice were intramuscularly immunized with DNA vaccine, saline and pcDNA3, respectively. The levels of anti-Ag85B antibody were examined with ELISA and the transcripts of IL-2, IL-4, IL-10 and IFN-gamma of spleen lymphocytes stimulated with recombinant Ag85B were tested with RT-PCR at the end of 6th week of inoculation. RESULTS: Compared with control group, the specific antibody levels against Ag85B were much higher and the IL-2 and IFN gamma transcripts were up regulated in the pcDNA3-Ag85B group. There was no change of IL-4 and IL-10 expressions in three groups. CONCLUSION: The pcDNA3 Ag85B stimulated the specific antibody products and induced the Th1 type cell immunity. PMID- 11930706 TI - [Molecular diagnostic techniques in identification of mycobacterial species]. PMID- 11930704 TI - [Systemic side effects of long-term treatment with low dose inhaled corticosteroids in children with asthma]. AB - OBJECTIVE: To observe the systemic side effects of low dose inhaled Beclomethasone dipropionate (BDP) in children with mild asthma. METHODS: 30 children with mild asthma were randomly divided into 3 groups to receive treatment with inhaled placebo (group A), BDP 200 micrograms/d (group B) and BDP 400 micrograms/d (group C) respectively. Bronchial hyperresponsiveness (BHR), height growth, bone mineral density (BMD), calcium and phosphate metabolism and hypothalamic-pituitary-adrenal axis (HPAA) function were measured. RESULTS: Inhaled BDP of 200 micrograms/d and 400 micrograms/d reduced BHR in mild asthmatic children and there was no significant difference between two groups [log(PD20-FEV1)]:(2.04 +/- 0.47) micrograms to (2.70 +/- 0.13) micrograms in group A and (1.94 +/- 0.46) micrograms to (3.15 +/- 0.18) micrograms in group B (P < 0.01). Serum osteocalcin, calcium, phosphate, alkaline phosphatase, basic cortisol and BMD didn't change significantly after BDP treatment in three groups (all P > 0.05) [In group A, B and C, concentrations serum osteocalcin were (29 +/ 12) micrograms/L, (22 +/- 6) micrograms/L, (31 +/- 11) micrograms/L, serum calcium: (2.49 +/- 0.11) mmol/L, (2.39 +/- 0.28) mmol/L, (2.20 +/- 0.35) mmol/L, serum phosphate: (1.8 +/- 0.6) mmol/L, (1.7 +/- 0.7) mmol/L, (1.5 +/- 0.4) mmol/L, radius BMD: (0.44 +/- 0.02) g/cm2, (0.42 +/- 0.05) g/cm2, (0.40 +/- 0.10) g/cm2, ulna BMD:(0.35 +/- 0.04) g/cm2, (0.36 +/- 0.08) g/cm2, (0.32 +/- 0.07) g/cm2, serum alkaline phosphatase: (410 +/- 113) U/L, (337 +/- 99) U/L, (351 +/- 122) U/L, serum basic cortisol: (350 +/- 86) nmol/L, (407 +/- 199) nmol/L, (365 +/- 71) nmol/L, lumbar spine (L4-5) BMD: (0.64 +/- 0.06) g/cm2, (0.59 +/- 0.08) g/cm2, (0.62 +/- 0.09) g/cm2 respectively]. Height growth had a trend of reducing after BDP treatment though not reaching statistical difference. Height standard deviation score (SDS): 1.1 +/- 0.7 to 1.2 +/- 0.9 in group A, 1.3 +/- 0.7 to 1.3 +/- 0.9 in group B and 1.1 +/- 0.7 to 1.0 +/- 0.7 in group C. Serum cortisol after ACTH stimulation reduced significantly in group C [(621 +/- 199) nmol/L to (482 +/- 97) nmol/L, P < 0.01]. CONCLUSION: The results of this study suggest that 200 micrograms/d BDP can reduce BHR significantly and has no detected systemic side effects in mild asthmatic children, and 400 micrograms/d BDP can reduce serum cortisol after ACTH stimulation. The long-term dose of BDP should be controlled to be less than 400 micrograms/d in children with mild asthma. PMID- 11930705 TI - [A study of heart rate variability in asthma]. AB - OBJECTIVE: To study the changes of autonomic nervous function in the young adult with asthma through the heart rate variability(HRV). METHODS: Two groups of young adults, 23 healthy volunteers and 69 asthmatic young adults were divided into three groups according to the severity of asthma. Twenty-four hours Holter monitored and heart rate variability analysed. RESULTS: The HF and pNN50 that showed the vagal tone in asthma subjects increased as compared with the normal group, while the LF and SDANN that mainly showed the sympathetic tone decreased(P < 0.01). These values were remarkable in the severe asthma group (P < 0.01). CONCLUSION: Even in mormal condition when patients are free of asthmatic attack, autonomic nervous function of asthmatics differ from that of normal young adults. PMID- 11930707 TI - [The clinical significance of Her-2/Neu in lung cancer]. PMID- 11930708 TI - [The orthopedic treatment of skeletal class III malocclusion with maxillary protraction therapy]. AB - OBJECTIVE: The aim of this study was to investigate the changes of skeletal anterior crossbite in early-stage after maxillary expansion and protraction therapy. METHODS: 40 chinese children with skeletal anterior crossbite were divided into two groups: the control group received no orthodontic treatment and the experimental group received maxillary expansion and protraction. The cephalometric analysis was used to evaluate the changes in both groups. RESULTS: In the experimental group, A point moved forward 3.5 mm (-1.75 mm in the control group). SNB, SNPg decreased and ANB, NP-PA increased. Protraction therapy to treat skeletal anterior crossbite in the middle and late stage of mixed dentition could influence craniofacial growth and development, such as accelerating forward growth of the maxilla, making mandible downward growth. CONCLUSIONS: Protraction therapy can achieve successful result in treatment of skeletal anterior crossbite in middle and late mixed dentition. PMID- 11930709 TI - [Expression of mRNA of osteocalcin in condylar cartilage of young SD rats after functional protrusion]. AB - OBJECTIVE: To study the temporal and spatial patterns of the expression of mRNA in condylar cartilage of young growing SD rats, in condition of different functional treatment time span. METHODS: SD rats are randomly divided into 3 groups: control, all-day group (wearing functional appliance for 24 hours a day) and 12-hour group (wearing appliance for 12 hours in daytime). After one week, animals are sacrificed and Osteocalcin mRNA expression is studied by in situ hybridization using Oligonucleotide probe, calculated by image processing and macroscopic and microscopic statistics. RESULTS: Osteocalcin mRNA is expressed in condylar cartilage of young growing SD rats and the acrophase is around 6:00 am. After wearing the functional appliance, the intensity in all-day group rises more than that in 12-hourgroup. CONCLUSIONS: Better results can be achieved in all-day group with functional appliance. PMID- 11930710 TI - [Effect of SP on intracellular free calcium concentration in cultured osteoblast]. AB - OBJECTIVE: To investigate the effect of SP on intracellular free calcium concentration (Ca2+)i in cultured osteoblast. METHODS: Effect of SP on intracellular free calcium was measured using cell culturing and laser scanning confocal microscopy with fluo-3/AM as a fluorescence probe. RESULTS: After treated with SP(10(-4) mol/L), 70% of the cells responded to it; the intracellular (Ca2+)i increased markedly [fluorescence intensity increased by (90 +/- 18)%]. CONCLUSIONS: These results suggested that calcium may have a role as a intracellular secondary messenger in the action of SP in osteoblast. PMID- 11930712 TI - [Culture and cryopreservation of human mandibular condylar cartilage cells]. AB - OBJECTIVE: To establish an effective way in cryopreserving mandibular condylar cartilage cells. METHODS: Human mandibular condylar cartilage (MCC) cells were harvested and amplified by microcarrier culture technique. MCC cells were preserve in liquid nitrogen. The biological properties of cryopreserved MCC cells were investigated. RESULTS: Cells from liquid nitrogen preservation kept their phenotypic, proliferative an metabolic characteristics of MCC cells in vivo. CONCLUSIONS: Cryopreservation of MCC cells can supply adequate qualified cells for repairing the defects of mandibular condyle cartilage and temporomandibular joint disk. PMID- 11930711 TI - [A study on the change of caries activity of children with fixed orthodontic treatment]. AB - OBJECTIVE: To investigate the change of caries activity in children with fixed appliance. METHODS: 30 children of permanent dentition were treated with edgewise appliance. CAT was used to test the degree of caries activity before the orthodontic treatment, at the first week, at the first month, at the third month and at the sixth month of appliance bonding respectively. RESULTS: Compared with pre-treatment, there was a significant difference of CAT at the first, third and sixth month of treatment; compared with the first week of treatment, there was also a significant difference of CAT at the third and sixth month of treatment. CONCLUSIONS: The CAT degree increases in children with fixed orthodontic appliance and these children are more sensitive to dental caries. PMID- 11930713 TI - [Alteration of immunological properties of bacterial lipopolysaccharide by calcium hydroxide treatment]. AB - OBJECTIVE: To study alteration of immunological properties of LPS by Ca(OH)2 treatment, and to establish an experimental model in which ThP-1 cell line was cultured in vitro to observe the alteration of immunological properties of LPS. METHODS: ThP-1 cell line was used to establish a culture model in vitro. Amount of TNF-alpha released from stimulated cells acted as the index on immunological properties of the stimuli and alteration of immunological properties of LPS by Ca(OH)2 treatment. RESULTS: Within the range of observed LPS concentrations (10 micrograms/L-10 mg/L), the original immunological properties of LPS decreased significantly after treatment by Ca(OH)2 for a week (P < 0.001). When pH > or = 12.30, the amount of TNF-alpha excreted by monocyte did not differ significantly from the control group (P > 0.05). CONCLUSIONS: After treatment by Ca(OH)2 for a week, the original immunological properties of LPS decreased significantly. High pH value of Ca(OH)2 played a critical role during the alteration of immunological properties of LPS. PMID- 11930714 TI - [The remineralization of enamel lesions by casein phosphopeptide-amorphous calcium fluoride phosphate in vitro]. AB - OBJECTIVE: To evaluate the remineralizing effect on artificial enamel subsurface lesions in vitro by 2% casein phosphopeptide-amorphous calcium fluoride phosphate complexes (CPP-ACFP) solution. METHODS: Twenty-eight enamel blocks were cut from human third molars, in which 2 lesions were prepared from each block. The 28 samples were randomly assigned to the four study groups, in which each group was incubated in remineralization solution for 1, 3, 5, 10 days respectively. After remineralization the samples were sectioned, microradiographied and the mineral content were determined by microdensitometry. RESULTS: The remineralizing solution replaced 9.19%, 14.27%, 29.07%, 38.45% of mineral lost after 1, 3, 5, 10 days respectively (y = 8.9316 x0.6347, R2 = 0.9322; y: remineralizing rate, x: time). Significant differences were found in remineralizing rate between any two groups using One-way Scheffe multiple comparison. CONCLUSIONS: The 2% CPP-ACFP solution could significantly remineralize the artificial enamel subsurface lesion in vitro. The remineralizing rates were increased with the experiment time (within 10 days). PMID- 11930715 TI - [The effects of otitis media with effusion and hearing loss on the speech outcome after cleft palate surgery]. AB - OBJECTIVE: To find out the exact effect of OME and hearing loss on the speech outcome post palatoplasty in cleft palate patients. METHODS: Otoscopy and tympanometry were performed in 18 cleft palate patients whose ages ranged from 5 to 23 years old and hearing thresholds were also tested before cleft palates repair. All patients were followed up from 3 to 18 months postoperatively. Articulation scores were evaluated and linear correlation analysis was performed between these results and their mean hearing thresholds for air conduction in speech frequencies before the palate closure. The incidences of the OME were compared among the groups with different articulation scores. RESULTS: The postoperative articulation scores correlate negatively (r = -0.504, P = 0.032 < 0.05) to the degrees of hearing loss. But the incidences of OME had no significant differences among the groups with different articulation scores. CONCLUSIONS: The hearing level preoperation is more important than tympanometric data on the foresight of the speech result postoperation in cleft palate patients. PMID- 11930716 TI - [The preliminary study on the oxide film of pure titanium treated by anodic oxidation]. AB - OBJECTIVE: To study the oxide film of pure titanium implant material treated by anodic oxidation and to learn the effect of anodic oxidation on it. METHODS: Six titanium sheets, which were ground, polished and pretreated, were treated by anodic oxidation at 10 v, 24 v or 40 v for 10 minutes and at 24 v for 10 minutes, 40 minutes or 2 hours. The microstructures and the corrosive-resistance of oxide films were studied by means of X-ray diffraction and electro-chemical corrosion. RESULTS: The results of X-ray diffraction showed that the titanic peaks of 2.55 and 2.34 inverted as potential went up. Not many disorderly peaks of titanium oxide were found. And all the peaks of 2.55 and 2.34, 1.25 and 1.23, 1.72 and 1.47 did the same when time went on. Disorderly peaks of titanium oxide first appeared more and then fewer. The open loop potential of the oxide film treated by anodic oxidation moved positively, and polarized current was a 100'th of that of natural oxide film. CONCLUSIONS: The oxide film with a certain color and crystallinity could be controlled by the potential and time. The corrosive resistance of the oxide film is greatly improved by means of anodic oxidation. It will be the next study how to utilize and adjust the rule of titanium crystal corrosion. PMID- 11930717 TI - [The influence of surface roughness on bacteria adhesion on titanium]. AB - OBJECTIVE: To investigate the correlation between surface roughness and bacteria adhesion. METHODS: Surface roughness of four abutments were determined by Talysurf/6p-120 topographer. Ra, Rz, Rp, Rv, Ry and Sm values were gathered. Commercial pure titanium and titanium alloy discs, S. mutans and A. viscosus were used in adhesion experiment. RESULTS: Ra values (in microns) of the tested abutments were: 0.1556, 0.2073, 0.6976, and 0.3811. No statistical difference was observed in bacterial quantity when Ra < 0.4528 microns and Rz < 3.3882 microns in adhesion experiment. CONCLUSIONS: The surface roughness of Ra < 0.4 micron and Rz < 3.4 microns is recommended for implant abutment. PMID- 11930718 TI - [The influence of two different casting temperatures on properties of the incorporated keeper]. AB - OBJECTIVE: To probe the clinical possibility of making incorporated keeper cap by high temperature casting. METHODS: The incorporated keepers were equally divided into two groups, and cast by applying medium and high temperatures respectively. The two groups were compared with the following items: the morphological changes and thickness of keeper's oxide film, the combining quality between keeper and alloy, and the magnetic retention force. RESULTS: The two different casting temperatures had the similar influence on the keeper's properties, such as the keeper's metallographical view, magnetic force (medium temperature casting group: 433.6 g; high temperature casting group: 433.8 g), and combining quality with alloy, except for the morphology and thickness of the keeper's oxide film. The oxide films of the high temperature casting group (surface: 7.59 microns combining surface of keeper and alloy: 6.17 microns) were thicker than those of the medium temperature casting group (surface: 4.65 microns; combining surface of keeper and alloy: 3.95 microns). CONCLUSIONS: The incorporated keeper could be made by using high temperature casting method instead of medium temperature one in clinic. PMID- 11930719 TI - [Evaluation clinical long-term curative effect and analysis of its influence factors after restoration of teeth with wedge-shaped defects]. AB - OBJECTIVE: To evaluate the long-term curative effect clinically and analysis of its influence factors after restoration of teeth with wedge-shaped defects. METHODS: Seventy five cases of 509 teeth with various degrees of dental wedge shaped defects were randomly divided into two groups--the group restored by Dyract Cement and the group restored by 3M-Vitremer. Evaluation was made with the modified USPHS evaluation criterion after restoration for 1,2,4 years respectively. RESULTS: The successful rate dropped down from 2 year successful rate 85.5% to 4-year 27.7%. The new wedge-shaped defects were presented at the cervical margin of the original restoration in 1.8% (2 years later) and 6.6% (4 years later). There were a lot of influence factors on long-term curative effect. The failure rate in the group with occlusal interference was significantly higher than that without occlusal interference. CONCLUSIONS: The clinical successful rates are affected by the value of occlusal force on the restored teeth and the physical properties of the restorative materials. It is suggested that the process of restoration of dental wedge-shaped defect should include obviating occlusal factors as well as to improve the properties of restorative materials. PMID- 11930720 TI - [Effects of change of crown contour on health of gingiva]. AB - OBJECTIVE: To study the effects of different excessive crown contours on the health of periodontium. METHODS: A total of 63 posterior teeth of 12 volunteers were randomly divided into three groups and each group was consisted of 21 teeth. The buccal surfaces of the teeth were overcontoured 0.2 mm, 0.5 mm, and 0.8 mm respectively. At the point of 1 week, 4 weeks and 8 weeks, gingival cervical fluid (GCF) were collected, GCF-aspartate aminotransferase (GCF-AST) and GCF alkaline phosphatase (GCF-ALP) levels were tested and the debris amount was assessed for three groups. RESULTS: The 0.8 mm group presented most amount of GCF, highest level of GCF-AST, GCF-ALP and most debris. The 0.5 mm group was the second. When the excess was 0.2 mm, all indexes showed no statistically significant change. CONCLUSIONS: Excessive crown contour is unfavourable to periodontium health. The more the teeth are excessively contoured, the more they impair periodontium. PMID- 11930721 TI - [The efficacy of the chlorhexidine chip following scaling and root planing (SRP) and compared to SRP alone]. AB - OBJECTIVE: To evaluate the clinical efficacy and safety of the adjunctive use of Chlorhexidine Chip (CHX; commercial name, Perio Chip) following scaling and root planing (SRP) in periodontitis. METHODS: One center, blinded, randomized, split mouth and active control study was designed. Sixty-five adult periodontitis patients were enrolled into the baseline after SRP. Each subject had at least one tooth with pocket depth of 5 mm or more and bleeding on probing (BOP) in each side of the mouth. Then the tooth was selected as a target and its parameters of probing depth (PD), attachment loss (AL), BOP, gingival index (GI), plaque index (PI) and staining index (SI) were recorded. Each side of mouth was randomly assigned to one of two treatments--drug placement after SRP or SRP alone. All the adverse events and parameters were recorded at time of 3-month, 6-month and meanwhile the patients received oral hygiene instruction and scaling. The same was conducted at 6-week, 4.5-month except for recording of parameters and scaling of the target teeth. RESULTS: Reduction of PD and gain of attachment at 6-month in group of SRP plus CHX (1.32 mm, 0.94 mm) were significantly higher than those in group of SRP alone (0.77 mm, 0.40 mm) (P < 0.001). Forty-four point six percent (44.6%) of patients reflected adverse reactions related to drug placement. Toothaches, the main reactions, were mild to moderate in nature and spontaneously resolved within 2-4 days. CONCLUSIONS: The chlorhexidine chip is indeed a safe and effective control-delivered drug for topical use when patient in his supportive periodontal therapy. PMID- 11930722 TI - [Intramandibular course and anatomic structure of the inferior alveolar nerve canal]. AB - OBJECTIVE: To study the intramandibular course and anatomic structure of mandibular canal. METHODS: The intramandibular course of mandibular canal and its dimension were measured in 15 adults mandibles with teeth; the relationship between blood vessel and nerve was observed in 4 fresh samples in which the artery was filled. RESULTS: The mandibular canal was close to the lingual side and the inferior margin of the mandible. The blood vessel lay above nerve in the mandibular canal. CONCLUSIONS: In implant operation performed according to normal anatomy, injury to inferior alveolar nerve may be avoided. Hemorrhage is the earliest manifestation suggesting that continuing drill may damage the inferior alveolar nerve. PMID- 11930723 TI - [Experimental study on effect of capping pulp with collagen]. AB - OBJECTIVE: To investigate the function of collagen on reparative dentin formation, collagen was used as capping pulp material in the teeth of minipigs. METHODS: The histopathologic examinations and ultrastructures were observed after capping for 2 weeks, 4 weeks and 13 weeks. RESULTS: After 2 weeks, the reparative dental masses were formed and the stellative cells appeared. After 4 weeks, the reparative dental bridges were formed and the degenerative cells appeared in matrix. After 13 weeks, the complete, think and dense dental bridges were observed. CONCLUSIONS: It is suggested that capping with collagen can promote formation of reparative dentine without stimulation to the pulp and that collagen can be used for capping. PMID- 11930724 TI - [Detection of HPV type 16, 18 infection and p53 protein overexpression in oral squamous cell carcinoma]. AB - OBJECTIVE: To study the action and interaction of HPV16, 18 infection and p53 protein overexpression in carcinogenesis of oral squamous cell carcinoma (OSCC). METHODS: Polymerase chain reaction (PCR) was used for detection of HPV16, 18 DNA, immunohistochemical technique (LSAB) for detection of p53 protein overexpression. RESULTS: In HPV-DNA analysis, HPV 16, 18 DNA were demonstrated significantly (P < 0.05) more often (48.89%, 22 of 45; 15 cases positive with HPV16-DNA and 7 cases positive with HPV18-DNA) in OSCC than in normal oral mucosa (negative). The frequency of p53 protein overexpression was 62.22% (28 of 45) in OSCC, while negative in normal oral mucosa (P < 0.01). The average survival time and survival rate of patients with HPV16, 18 infection and p53 overexpression was shorter than that of patients without HPV infection (P > 0.05) and without p53 overexpression (P < 0.05). CONCLUSIONS: The result suggests that HPV16, 18 infection plays a significant role in carcinogenesis of OSCC. p53 mutation is a frequent genetic event in OSCC. HPV status and p53 protein overexpression partially affect the average survival time and survival rate of OSCC patients. PMID- 11930725 TI - [Applied study on computerized quantitative facial nerve functional estimating system]. AB - OBJECTIVE: To prove the clinical applied value of quantitative facial nerve functional estimating system (QFES) by comparing the estimating results of the computerized QFES with manual QFES. METHODS: 50 volunteers with normal facial nerve function were used. Every volunteer was taken 7 photos with digital camera, which including static, and forehead wrinkle, eyebrows knit tight, eyes closed tight, nose wrinkle, smile, pout and mouth open to maximal seven expressions. The facial nerve function was estimated by computerized QFES, and the estimating results were compared with manual estimations by QFES in our study during 1991 1994. RESULTS: All estimating indices in this study were in the range of 0.70 0.90 as well as the manual estimating results. However, the stand deviation in this study showed significantly smaller than the previous study. CONCLUSIONS: QFES is an ideal quantitative facial nerve functional estimating system, after computerization the efficiency is increased and the systemic error is decreased. It will benefit to apply computerized QFES in the facial nerve function estimating fields. PMID- 11930726 TI - [The adenovirus-mediated HSV-TK/GCV suicide gene system in the treatment of tongue carcinoma cell line]. AB - OBJECTIVE: To study the effects of replication-defective recombinant adenovirus mediated herpes simplex virus thymidine kinase (AdCMV HSV-TK)/ganciclovir (GCV) suicide gene system on tongue carcinoma cell line. METHODS: Replication-defective recombinant adenovirus containing CMV promoter were propagated in 293 cells, purified by cesium chloride gradient and titrated by tissue culture infectious doses50 (TCID50) methods. Infectivity test was used to determine the efficiency of adenoviral transduction into tongue carcinoma cell line (Tca8113 cell line) and RT-PCR was applied to detect the expression of TK gene after infecting Tca8113 cells. The killing effects and bystander effects of HSV-TK/GCV system on tongue carcinoma cells were detected with MTT assay. RESULTS: The adenoviral titration was about 2 x 10(10) plaque-forming units(PFU)/ml. The efficiency of adenoviral transduction into tongue carcinoma cells was more than 70 percent when multiplicity of infection (MOI) was more than 20. The recombinant adenovirus expressed TK gene after infecting Tca8113 cells in vitro. Adenovirus-mediated HSV TK/GCV suicide gene system had killing effects on Tca8113 cells and these effects were related with time and GCV dose. The killing effects were poor after treated for 3 days and increased for 5 and 7 days. There had a good killing effect only when MOI was more than 20. When MOI was 80, GCV 5 x 10(-4) mol/L and treatment for 7 days, the viability of cells is only 4%. The bystander effect of this system on Tca8113 cells is poor. CONCLUSIONS: Only when most of the tongue carcinoma cells were transferred by AdCMV HSV-TK, HSV-TK/GCV suicide gene system has good killing effect on tongue carcinoma cell line. PMID- 11930727 TI - [Relationship between expression of inducible nitric oxide synthase and of VEGF in oral carcinoma]. AB - OBJECTIVE: To study the expression of inducible NOS in oral carcinoma and it's relationship with the expression of VEGF, MVD and lymphatic metastasis. METHODS: With 9 cases of normal oral mucosa as control, the expression of iNOS in 41 cases of oral carcinoma was evaluated by immunohistochemistry SP staining. Meanwhile MVD and the expression of VEGF were also detected. RESULTS: The expression rate of iNOS in 41 cases was 63.41%, and the rate between N+ group and N- group was significantly different (P < 0.01). There were significant difference of MVD (P < 0.001) and of the expression of VEGF(P < 0.05) among groups of different iNOS expression level. CONCLUSIONS: The expression of iNOS in oral carcinomas is high, and it is closely related to angiogenesis and lymphatic metastasis. The expression of iNOS probably takes part in VEGF's angiogenic effect. PMID- 11930728 TI - [Antimetastatic effects of arginine-aspartate on salivary adenoid cystic carcinoma in vitro]. AB - OBJECTIVE: To test the effects of arginine-aspartate (RD) on metastasis of salivary cystic carcinoma (SACC). METHODS: Highly metastatic SACC cell line SACC LM was used in the study. The effect of RD on tumor cell-ECM adhesion was assayed by MTT technique. The effect of RD on invasion and migration of SACC-LM were evaluated using modified boyden chamber. RESULTS: RD inhibited tumor cell fibronectin adhesion at 5 mg/L concentration, but no effects on tumor cell laminin and tumor cell type IV collagen adhesion. RD inhibited the invasion at 1, 5, 25 mg/L, and inhibited the migration only at 25 mg/L. CONCLUSIONS: The result shows that the RD has an antimetastatic effect on SACC-LM cells in vitro. PMID- 11930729 TI - [Prognostic significance of heat shock proteins 70 and p27 protein in patients with mucoepidermoid carcinoma]. AB - OBJECTIVE: To study the expression of heat shock proteins 70 (HSP70) and p27 in mucoepidermoid carcinoma (MEC) in relation to biological behaviour and prognosis. METHODS: Immunohistochemical LSAB method was used to evaluate the expression of HSP70 and p27 in 44 MEC. 10 Pleomorphic adenomas (PAs) were used as controls. RESULTS: The expression of HSP70 was significantly higher in MEC than in PAs, while p27 expression was significantly lower in MEC than in PAs. There was a negative correlation between HSP70 and p27 expression. Compared with clinicopathologic features, the expression of HSP70 was related to histologic grade, clinical stage and tumor size, and p27 expression correlated negatively with histologic grade, clinical stage, tumor size and lymph node metastases. Multivariate analysis revealed that Hsp70 expression was a strong prognostic indicator of MEC. CONCLUSIONS: HSP70 should be considered an independent prognostic indicator of MEC and p27 might favour prognosis of MEC. PMID- 11930730 TI - Current status of aortic aneurysm surgery in Hong Kong. AB - OBJECTIVE: To determine the epidemiology and the status of open and endovascular aortic surgery for aortic aneurysm in Hong Kong. METHODS: Three separate data sources were obtained: (1) the Hong Kong Hospital Authority discharge statistics for 1999 and 2000; (2) a survey on aortic aneurysms in public hospitals conducted by the working group of vascular surgery; and (3) the department of surgery, University of Hong Kong Medical Center aortic aneurysm database. The disease pattern, distribution as well as audit of operative mortality was determined. RESULTS: Aortic aneurysm ranked tenth as the leading causes of death in Hong Kong, and the incidence is increasing. Almost 800 new cases were diagnosed each year, with 10% presenting as rupture, but the death rate for ruptured aneurysms was 80%. About half of all operations on aortic aneurysms was performed for rupture, and a significant number of newly diagnosed patients were not receiving surgery. In experienced centers, the operative mortality for elective and ruptured aneurysm have improved to 2% and 38% in recent years. A growing interest and number of endovascular repair operations were performed which has led to some concerns on patient selection and follow up. CONCLUSIONS: Similar to a worldwide trend, aortic aneurysm in Hong Kong is diagnosed more frequently. With the relatively high mortality for ruptured aneurysms, effective diagnosis and elective surgery on patients with aortic aneurysms in experienced vascular centers remained the best treatment. Since a majority of aneurysms remained untreated, patient and physician education is of paramount importance. PMID- 11930731 TI - [Fractionated-clamping for thoracoabdominal aortic aneurysm repair: report of 13 cases]. AB - OBJECTIVE: Fractionated-clamping was applied to aneurysm repair in patients with thoracoabdominal aortic aneurysm(TAA), and its effects in decreasing operative mortality and severe complications such as renal failure and hemiplegia were discussed. METHODS: Using improved shunting and cross-clamping technique, we repaired 13 thoracoabdominal aortae and their branches (9 males and 4 females). TAA Crawford type: type I one case, type II two cases, type III two cases and type IV three cases. TAA dissection Debakey type: type I one case, type III four cases (including 2 of ruptured aneurysm) and aorta stenosis one case. RESULTS: Twelve procedures were performed successfully. One patient died of cardiac attack immediately before accomplishment of the operation with a operative mortality rate of 7.7% (1/13). Operative complications included acute necrotizing pancreatitis (1 case), ARDS (1), thoracic cavity bleeding (2), hemiplegia (1), and acute renal failure (1). The incidence rate of complications was significantly lower than that reported elsewhere. CONCLUSIONS: Fractionated clamping for thoracoabdominal aortic aneurysm repair is a renovation on the basis of Crawford procedure and aorta bypass method. Clinical results demonstrated that the procedure decreases the surgical mortality and the incidence rate of dangerous complications. Fractionated clamping for aorta replacement is a reasonable procedure under general anesthesia at normal temperature. PMID- 11930732 TI - [Early mortality and complications of abdominal aortic aneurysm repair: analysis of 186 cases]. AB - OBJECTIVE: To study the early mortality and major complications of infrarenal abdominal aortic aneurysm (AAA) repair. METHODS: One hundred and eighty-six cases of infrarenal AAA, which underwent surgery in our hospital between January 1988 and December 2000, were selected. Early mortality and major complications were rated, and preoperative cardiac, pulmonary and renal condition, age and surgical impact were taken into account while evaluating the risk factors of early mortality and major complications. RESULTS: The early mortality was 5.0% for elective surgery and 57.1% for ruptured surgery. The early major complication rate for AAA repair was 18.4% in total, and 10.6%, 11.2%, 2.8%, 1.1% for cardiac complication, pulmonary complication, acute renal failure, stroke and hepatic insufficiency respectively. Coronary heart disease significantly predisposed the patients to the development of cardiac complications (chi 2 = 19.737, P < 0.01) while hypertension had no such effect (chi 2 = 1.870, P > 0.05). Abnormal pulmonary function, smoking and PaO2 less than 80 mmHg were responsible for postoperative pneumonia(chi 2 = 4.051, 5.885 and 5.162, P < 0.05), but not for adult respiratory distress syndrome(chi 2 = 0.127, 0.916 and 1.067, P > 0.05). Preoperative renal function was not related postoperative acute renal failure(chi 2 = 0.404, P > 0.05). Age more than 70 years or operation spanned more than 5 hours increased major complications(chi 2 = 16.119 and 10.163, P < 0.01) and early mortality(chi 2 = 16.119 and 10.163, P < 0.01). CONCLUSIONS: Multiple system organ failure is the leading cause of early mortality of AAA repair. Cardiac and pulmonary complications are commonly seen after AAA repair. Preoperative cardiac, pulmonary condition, age and surgical aspects greatly influence the early mortality and major complications. PMID- 11930733 TI - [Management of arterial occlusive diseases of abdominal aorta and iliac artery: experience with 267 cases]. AB - OBJECTIVE: To study the surgical treatment of infrarenal aortic and iliac arterial occlusive disease. METHOD: We analyzed 267 patients with arterial occlusive diseases of the abdominal aorta and iliac artery retrospectively. RESULTS: All the patients were treated by surgery including aortic-iliac bypass in 145 patients, iliac-femoral bypass in 40, femoro-femoral bypass in 45, and axillo-femoral bypass in 37. Their total effective rate and perioperative mortality were 96.5% and 3.5%, respectively. Of the 267 patients, 178 were followed up. The patency rate of prosthesis was 82.0%. CONCLUSIONS: Extra anatomical bypass is a procedure of choice for high-risk patients, especially for those chronic patients with coronary heart disease, hypertension, cerebral arterial atherosclerosis, and brain infarction. PMID- 11930734 TI - [Surgical treatment of abdominal aortic aneurysms: report of 482 cases]. AB - OBJECTIVE: To improve the safety of abdominal aortic aneurysm (AAA) repair. METHOD: The experience of 482 cases of AAA in surgical treatment and endovascular graft exclusion were reviewed from January 1960 to March 2001. RESULTS: Retroperitoneal approach and minimal incision was used for AAA operation. New methods for control of the "neck" of aneurysm, aneurysmectomy and endovascular graft exclusion were applied. The danger of AAA repair obviously decreased. The operative mortality was 5.2% and the five-year survival rate was 74.4%. CONCLUSION: The improvement of surgical and anesthetic techniques made AAA resection rapid and safe. PMID- 11930735 TI - [Endovascular stent-grafting of descending thoracic aortic lesions]. AB - OBJECTIVE: To discuss the feasibility of endoluminal stent-grafting for descending thoracic aortic lesions. METHODS: 12 patients underwent stent-grafting of the thoracic aorta with Talent (11 patients) and Vanguard (1) device. Endovascular technique was used in 12 patients with descending thoracic aortic aneurysms (5 patients), Stanford B dissections (6), and pseudoaneurysm (1). 13 straight stent-grafts were inserted through 18-24 F theaths via a femoral arteriotomy under local (2 patients) or general (10) anesthesia. RESULTS: The procedure was technically successful in all patients without aneurysm rupture, paraplegia, and organs or limbs ischemia. There were 3 early endoleaks. Follow-up showed that 4 of 5 aortic aneurysms were excluded completely and one endoleak was changed into persistent. Four entries of 6 patients with stanford B dissection were excluded and 2 early endoleaks were sealed during one month after operation. Four false lumen were filled completely and 2 false lumen were filled partly by thrombosis. CONCLUSION: Descending thoracic aortic stent-grafting is a safe, highly successful and feasible alternative to conventional surgery. The long-term result remains unclear, especially that of the endovascular repair of Stanford B dissection. PMID- 11930736 TI - [Clinical study of detenia cecal-ascending colon continent cutaneous urinary reservoir]. AB - OBJECTIVE: To construct a good continent urinary diversion which is easy to be performed and has a low incidence of complications. METHODS: 26 cases of bladder cancer were given radical cystectomy before the cecal-ascending colon was excluded and 15-20 cm of the cecal-ascending colon was isolated. The colon teniae were then incised at the interval of 0.5-1.0 cm to construct the detenia cecal ascending colon continent urinary diversion open to the umbilicus. RESULTS: All the patients were followed up for 21.1 +/- 10.1 months. Reliable continence was achieved in all with a low incidence of complications. The capacity of the reservoirs reached 350-600 ml 6 month after operation. Self catherizations were carried out every 3 to 6 hours, Urodynamic data showed a mean maximum filling pressure of 58.7 +/- 24.5 cmH2O, and a mean maximum urethral (efferent) closure pressure of 104.3 +/- 33.8 cmH2O. CONCLUSION: Detenia cecal-ascending colon continent urinary diversion is an ideal method. PMID- 11930737 TI - [Enhanced continent mechanism of the tapered ileum in continent urinary reservoir]. AB - OBJECTIVE: To construct a reliable continent tube that is easy to catheterize and is surgically simple. METHODS: From October 1999 through March 2001, 20 patients underwent a procedure in which ileal segment was tapered as an efferent tube and the partial efferent tube was placed between the back surface of the rectus muscle and the wall of the ileal pouch. The internal orifice of the tapered ileum was anastomosed to the ileal pouch and its external orifice of the tapered ileum was anastomosed to the umbilicus. Urodynamic study of the efferent tubes and pouch was done 1.5 to 3 months and 6 to 17 months after operation. RESULTS: The stoma was easily catheterized with a 16 F catheter in all patients. One patient died of heart disease 55 days after the operation, while 18 of the remaining 19 were completely continent day and night. At 1.5 to 3 months, the urodynamic study of the efferent tubes showed the maximum close pressure with a full pouch of 46 124 cmH2O(91.53 +/- 17.21), and when the pouch was empty it was 34-84 cmH2O(66.68 +/- 11.60). The difference in the mean maximum closure pressure in full and empty pouches was statistically significant (t = 10.59, P < 0.01). At 6 to 17 months, urodynamic study was performed in 12 patients, the maximum closure pressure in the efferent tube was 77 to 154 cmH2O (100.92 +/- 20.88) when the pouch was filled with saline. When the pouch was empty, it was 56 to 115 cmH2O (74.08 +/- 14.59). The difference in the mean maximum closure pressure in full and empty pouches was statistically significant (t = 8.54, P < 0.01). Reservoir capacity was 360 to 750 ml (455 +/- 110.74). When it was filled to the maximum, the reservoir pressure was 16 to 35 cmH2O (23.17 +/- 5.82). There was no contractive wave in filling in any patient. CONCLUSIONS: This study indicates that the continent mechanism of the tapered ileum can be greatly enhanced by fixing it between the abdominal and pouch walls. This maneuver also provides easy catheterization and surgical simplicity. PMID- 11930738 TI - [Orthotopic detenia cecal-ascending colon urinary reservoir: report of 18 cases]. AB - OBJECTIVE: To improve the technique of detenia cecal-ascending colon continent urinary reservoir for farther improving the life quality of the patient. METHODS: Orthotopic detenia cecal-ascending colon urinary reservoir was carried out by complete resection of all the tenia and multiple transverse incision of colonic circular muscular layer for 18 patients with bladder cancer underwent radical cystectomy. RESULTS: The patients have been followed up for 6-20 months. All were completely continent during the day and 13 patients got complete continence at the night. Only 3 patients had nocturnal incontinence. The capacity of the urine reservoir was 410-520 ml. The maximum intrareservoir pressure was 39-60 cmH2O, while the post-micturition residual volume was 15-46 ml and the maximum flow rate was 12-28 ml/per second. No evidence of ureter reflux and ureteral obstruction was observed and serum electrolytes were normal in all patients. CONCLUSIONS: Orthotopic detenia cecal-ascending colon urinary reservoir shows good clinical and functional results, and the simple technique, exerts low pressure in the reservoir and produces minimal complications. PMID- 11930740 TI - [Prospective double-blind randomized trial of cryoanalgesia: experimental and clinical study]. AB - OBJECTIVE: To alleviate and prevent postthoracotomy severe chest pain. METHODS: In dogs, intercostal nerves were freezed at -50 degrees C at different times. Pathological changes and repair process of the nerves were observed while identifying the best freezing time that could be used clinically. Prospective double-blind randomized trial in 200 patients who had undergone thoracotomy was carried out from January 1996 to June, 2000. Cryoanalgesia(Cryo group) and control group consisted of 100 patients. Before closing chest, 4 routes of costal nerves (incision, above and below incision, and one for inserting drainage tube) were freezed for 1.5 minutes separately. Postoperative incision pain was recorded by VAS(visual analogue scales) including recording the dosage of narcotics, pulmonary function on 3, 7, 15, 30, 90 postoperative days. RESULTS: Pathological changes of the axons and myelin sheaths progressed along with freezing time from 30 seconds to 2 minutes, but recovered in about one month. As the best freezing time, 1.5-minute freezing provided effective analgesia and quick recovery. Clinical application showed strong analgesia effect. VAS score of the cryo-group and control group was 2.29, and 7.24 (P < 0.001) respectively. In the cryo-group, no pain (0 degree) was observed in 22 patients (22%), very slight pain (1-2 degrees) in 44(44%), slight pain (3-4 degrees) in 20 (20%), and moderate pain (5 6 degrees) in 8 (8%). The total effective rate (0-6 degrees) was 94%. Dosage of dolantin in the cryo- and control groups was 41.3 mg and 185 mg, respectively(P < 0.001). The changes of pulmonary function for the two groups were not significant. CONCLUSION: Freezing intercostal nerves during thoracotomy can effectively prevent postoperative chest pain. The pathological changes of nerves are reversible. PMID- 11930739 TI - [Surgical treatment of great vessel injuries of thoracic outlet]. AB - OBJECTIVE: To summarize the experience in surgical treatment of great vessel injury of the thoracic outlet. METHODS: Thirty-two cases (24 men and 8 women aged 2-48 years) treated by surgery were collected during the period of 1970-1998. Sharp injury, blunt injury or decelerated injury was the causes. The vessels of injury included the common carotid artery, subclavian artery and accompanying veins. Emergency suture or repair of the artery with polyester patch was performed in 15 patients, aneurysmectomy in 6, resection of vessels with end-to end anastomosis in 5, Gore-Tex vessel reconstruction in 2, and transplantation of the great saphenous vein in 3. RESULTS: Twenty-four patients (75%) were cared and 7 (21.8%) died. CONCLUSIONS: In the early period, patients with great vessel injury of the thoracic outlet died from acute bleeding, cerebral ischemia or combined multiple injuries, in the later period, however, false aneurysm develop. The operation is appropriate to the recondition of damaged cut, the effect of evolution in false aneurysm is approving. For large damaged cut or for bigger vessel injuries, the repair with polyester patch, Gore-Tex of blood vessel reconstruction or the transplantation of the great saphenous vein must be used. PMID- 11930741 TI - [Microsurgical treatment of apical region by combined orbital pterygoid approach]. AB - OBJECTIVE: To study microsurgical treatment of orbital apical region tumor by combined orbital pterygoid approach. METHOD: In 21 cases, manifestations and surgical methods were analyzed retrospectively. The surgical approach and procedure were discussed. RESULTS: All the 21 cases were treated by combined orbital pterygoid approach. The tumors were removed completely in 18 cases, and partially resected in 3. There was no death. CONCLUSIONS: This kind of mass is difficult to explore by either transorbital or transcranial approach because of its position. Combined orbital pterygoid approach provides better access and exploration. Appropriate surgical approach, hemostasis and gentle microsurgical manipulation are the key to reducing postoperative complications. PMID- 11930742 TI - [Shoulder pain after laparoscopic cholecystectomy]. AB - OBJECTIVE: To detect the mechanism of shoulder pain after laparoscopic cholecystectomy(LC). METHODS: Ninety patients who had receive LC were randomly divided into three groups (each 30 patients): group A, no management after LC; group B, absorption of residual CO2 after LC; group C, O2 insufflation for three times followed by complete absorption of CO2 after LC. PO2 and PCO2 before and after LC were recorded. Postoperative shoulder pain was assessed with shoulder pain's rate and level. RESULTS: The PO2 difference of group C between before and after LC was significant as compared to groups A and B(P < 0.05). Postoperative shoulder pain developed in 13 patients (43.33%) of group A, in 8 (26.67%) of group B, and 21 (70%) of group C (P > 0.05 A vs B; P < 0.05 A vs C; and P < 0.01 B vs C). The shoulder pain's level was higher in group C than in groups A and B. CONCLUSIONS: Direct irritation to the diaphragm due to the residual CO2 pneumoperitoneum is not the main cause of shoulder pain. The excessive traction of the triangular ligament and over-stretching of the diaphragmatic fibers due to insufflation are the main cause of shoulder pain. PMID- 11930743 TI - [Cox proportional risk model analysis on the affecting factors of liver transplantation]. AB - OBJECTIVE: To analyze the relative factors affecting the survival time of liver transplantation and identify the main factors for the purpose of supply the reference evidence to the clinic. METHODS: The clinical data on preoperative condition, interoperation and liver function in 116 out of 128 recipients were quantified by the cox proportional risk model after liver transplantation. Their variables were calculated and analyzed by cox regression under the SAS 6.12 software. RESULTS: Z14 (FFP) was obvious at alpha = 0.05. After deleting the non obvious factors from the model, regression coefficient, standard errors, wald, P and RR were 0.065, 0.024, 7.493, 0.006 and 1.067 respectively. There was no difference of survival time between living liver transplantation and cadaver liver transplantation. The relative dangerous degree in cadaver liver transplantation was 0.340 as much as that in living liver transplantation. CONCLUSIONS: Living liver transplantation shortens the waiting time of liver transplantation and alleviates patient's suffering. It suits for children and adult and supports a selective liver transplantation. PMID- 11930744 TI - [Retrograde interlocked intramedullary nailing of fresh humeral shaft fractures]. AB - OBJECTIVE: To report the experience in retrograde intramedullary nailing of fresh humeral shaft fractures. METHOD: From February 1999 to June 2000, 32 humeral shaft fractures were treated with retrograde intramedullary nailing. All fractures were fresh and closed. The entrance point was on the proximal boarder of the olecranon fossa. Nails were locked distally and proximally. RESULTS: All fractures healed within 3 months and showed good function of shoulder and elbow joints. Radial palsy occurred in 1 patient post-operatively. CONCLUSION: Retrograde intramedullary nailing of humeral shaft fracture is one of the best treatments. Its advantages include no disturbance to the rotator cuff, stable fixation of the fractures, less soft tissue injury and fast recovery of the functions of near shoulder and elbow joint. PMID- 11930745 TI - [Clinical application of TRAFIX internal fixation system in correction of scoliosis]. AB - OBJECTIVE: To evaluate the results of TRAFIX internal fixation system in correction of adolescent scoliosis. METHODS: From October 1997 to June 1999, 27 patients with idiopathic scoliosis received spinal instrumentation with TRAFIX system. Their average age was 14.2 years (from 11 to 21). Four patients of the 27 underwent anterior release, while 1 patient accepted revision approach. The average follow up time was 26 months (13-37 months). RESULTS: The measurements for primary coronal deformity before and after surgery were 65 degrees (42 degrees-110 degrees) and 29.2 degrees (3 degrees-64 degrees) respectively. The average curve correction was 55.4%. The average fused segments were 11.5 vertebrae. The distance between the center of apex and C7 plumb line was 52.1 mm before operation (25-94 mm) and 28.5 mm after surgery (4-62 mm). CONCLUSION: The TRAFIX internal fixation system provides three-dimensional correction with refinement, convenient and reliable fixation. PMID- 11930746 TI - [Reconstruction of heel: long-term follow up of 12 cases]. AB - OBJECTIVE: To study long-term follow up result of fibula transplantation to reconstruct partial or complete defect of the heel. METHODS: Heel defects were reconstructed by means of reverse transferring of combined vascular pedicled fibulaflap. The calcaneus was repaired by two segments of the fibula. Skin defects were covered by skin flap, and sensation was restored by anastomosing the sural nerve and medial pedal nerve. The residual cavity was filled by muscles surrounding the fibula to reconstruct the appearance of foot. RESULTS: Four patients with complete heel defect and 8 with partial defect were operated on successfully. Two to 13 years(average 5.2 years) follow-up showed that the function and appearance of foot were normal and adaptive changes happened in the tissue of foot. The fibula was getting into a real cal caneous gradually. Besides this, the skin was thickened and the sensation was restored with the two-point discrimination of 0.8 to 3.0 cm. All patients resumed physical work. Comprehensive evaluation revealed that 11 patients were excellent and 1 was good in their functional recovery. CONCLUSION: The results suggested that this operation is ideal for reconstructing heel defect. All kinds of tissues in heel can be reconstructed and more satisfactory function and appearance can be obtained through adaptive heel reshaping along with long-term walk. PMID- 11930747 TI - [Establishment and biologic characterization of human hepatocellular carcinoma cell line HCC-9903]. AB - OBJECTIVE: To establish a new hepatocellular carcinoma (HCC) cell line and provide a new and ideal experimental model. METHODS: The cell line designated as HCC-9903 was derived from a surgical specimen of HCC. After primary culture, its biologic characteristics were studied under light and electron microscope and by determination of doubling time and growth curve, culture in soft agar, flow cytometry(FCM), chromosome analysis, detection of AFP, HBV and telomerase, agglutination test of concanavalin (ConA) as well as inoculation in nude mice. RESULTS: The first passage was found on the 29th day. Morphological study demonstrated its malignant characteristics. Chromosome analysis revealed a chromosome number ranging from 49 to 135, with a model number of 52 to 67. Aberrant chromosomes 1 q(i) and t (6,11) were its chromosome markers. The cell line produced tumors after subcutaneous transplantation into nude mice. CONCLUSION: According to the rules of establishing a cell line in vitro, HCC-9903 has been proved to be a new cell line of human HCC carcinoma, and more experimental methods have been used to study its biological characteristics. PMID- 11930748 TI - [Effect of PCNA antisense oligooxynucleotides and VEGF antisense oligoxynucleotides on growth of hepatocellular carcinoma transplanted in nude mice]. AB - OBJECTIVE: To study the inhibitory effect of PCNA antisense oligonucleotides or/and VEGF antisense oligonucleotides on growth of human hepatocellular carcinoma in nude mice. METHODS: Human hepatocellular carcinoma transplanted subcutaneously in nude mice model was established, and subsequently was divided into four groups: PCNA-ASODN group, VEGF-ASODN group, combined group and control group. Different treatments were given respectively. The weigh and volume of subcutaneous tumors was measured, and the morphological changes of tumor cells was observed under microscope. RESULTS: The growth inhibitory rate in the PCNA ASODN group was 72.8%, 44.9% in the VEGF-ASODN group, and 87.2% in the combined group. The combined group which was treated with PCNA-ASODN and VEGF-ASODN showed more effective inhibitory than the PCNA-ASODN group or the VEGF-ASODN group respectively. CONCLUSIONS: The growth of human hepatocellular carcinoma in nude mice can be inhibited by PCNA antisense oligonucleotides and VEGF antisense oligonucleotides respectively. Better inhibitory effect can be obtained if PCNA antisense oligonucleotides and VEGF antisense oligonucleotides are combined in use. PMID- 11930749 TI - [Intracellular signal transduction pathways of IL-1 beta-induced cytotoxicity on primary cultured rat hepatocytes]. AB - OBJECTIVE: To investigate the intracellular signal transduction pathways of IL-1 beta-induced cytotoxicity on primary cultured rat hepatocytes. METHODS: All studies were based on primary cultured hepatocytes from male Wistar strain rats (5 to 7 weeks old, 170 to 230 g). The activity of lactose dehydrogenase (LDH) was detected by colormetric assay. JNK and p38 kinase were analyzed by Western blot assay. The AP-1 activity was evaluated by electrophoretic mobility shift assay. RESULTS: IL-1 beta significantly increased LDH release. LDH activity in group IL 1 beta and controls was 21.9% +/- 3.6% and 11.0% +/- 1.8% respectively, P < 0.01). IL-1 beta induced cytotoxicity via activating JNK pathway and transcription factor AP-1 and the simultaneous activation of p38 kinase pathway negatively regulated this process. CONCLUSION: IL-1 beta induces cytotoxicity in primary cultured rat hepatocytes via activating JNK pathway and transcription factor AP-1. PMID- 11930750 TI - [Relationship between expression of IL-1 beta mRNA and rejection of heart allograft]. AB - OBJECTIVE: To detect relationship between expression of IL-1 beta mRNA and allograft survival time. METHODS: A simplified heterotopic heart transplantation model and a new reliable semi-quanitative RT-PCR technique were used. Inbred SD and Wistar rats were used as donors and recipients respectively in allogeneic transplantation. In synogeneic transplantation, both the donor and recipient were Wistar rats. Thirty allografts and thirty isografts were assessed daily by palpation to observe grafts' viability. In every time point, six allografts and six isografts were used to detect IL-1 beta mRNA expression. The allografts and isografts were harvested on postoperative days 1, 3, 5, 7, 9, 11. After the total RNA was prepared, the control primer and target primer were amplified in the same tube. The average ratio of the target products pixel volume to primer's was calculated, by which the coordinates chart was made to detect the expression level of IL-1 beta mRNA directly. RESULTS: The levels of the IL-1 beta mRNA expression in allografts were much higher than those in isografts. The peak of elevated level of the IL-1 beta mRNA expressions appeared on the 1st day post transplantation 1st day, and prior to the peak of the allografts dead time. CONCLUSIONS: The experiment suggested that the IL-1 beta may play an important role in rejection of heart graft. The determination of IL-1 beta mRNA expression may serve as a useful immunologic monitoring protocol, and some administrations against IL-1 beta may be used to prevent and treat rejection. PMID- 11930751 TI - The costs of penance. PMID- 11930752 TI - Let priests marry and ordain women. This is a time for radical change. PMID- 11930753 TI - [Diagnostic and therapeutic guidelines (algorithms) in the treatment of schizophrenia. Recommendations of the Croatian Society for Clinical Psychiatry of the Croatian Medical Association]. AB - This paper presents guidelines for the diagnosis and treatment of schizophrenia. Diagnostic guidelines are based on ICD-10 criteria for diagnosis of schizophrenia. Treatment guidelines represent a synthesis of some recently published international algorithms and our own experiences. The guidelines were presented at the seminar organized by the Croatian Association for Clinical Psychiatry and represent professional recommendations of the Association for Diagnostics and Treatment of Schizophrenia. The treatment guidelines include psychopharmacological recommendations for the selection and dosage of antipsychostics, guidelines for psychosocial interventions and psychotherapy. The treatment of the first episode, exarcerbation, relapse of illness, as well as maintenance therapy, treatment resistance and side effects are included. PMID- 11930754 TI - [Effect of inoculum size on sensitivity and specificity of the double-disk synergy test for the detection of wide-spectrum beta-lactamases]. AB - The plasmid-mediated extended-spectrum beta-lactamases (ESBL) confer resistance to oxymino-cephalosporins, such as cefotaxime, ceftazidime, and ceftriaxone and to monobactams such as aztreonam. It is well known fact that ESBL producing bacteria exhibit a pronounced inoculum effect against broad spectrum cephalosporins like ceftazidime, cefotaxime, ceftriaxone and cefoperazone. The aim of this investigation was to determine the effect of inoculum size on the sensitivity and specificity of double-disk synergy test (DDST) which is the test most frequently used for detection of ESBLs, in comparison with other two methods (determination of ceftazidime MIC with and without clavulanate and inhibitor potentiated disk-diffusion test) which are seldom used in clinical laboratories. The experiments were performed on a set of K. pneumoniae strains with previously characterized beta-lactamases which comprise: 10 SHV-5 beta-lactamase producing K. pneumoniae, 20 SHV-2 + 1 SHV 2a beta-lactamase producing K. pneumoniae, 7 SHV 12 beta-lactamase producing K. pneumoniae, 39 putative SHV ESBL producing K. pneumoniae and 26 K. pneumoniae isolates highly susceptible to ceftazidime according to Kirby-Bauer disk-diffusion method and thus considered to be ESBL negative. According to the results of this investigation, increase in inoculum size affected more significantly the sensitivity of DDST than of other two methods. The sensitivity of the DDST was lower when a higher inoculum size of 10(8) CFU/ml was applied, in distinction from other two methods (MIC determination and inhibitor potentiated disk-diffusion test) which retained high sensitivity regardless of the density of bacterial suspension. On the other hand, DDST displayed higher specificity compared to other two methods regardless of the inoculum size. This investigation found that DDST is a reliable method but it is important to standardize the inoculum size. PMID- 11930755 TI - [Arthroscopic meniscectomy in the General Hospital of Pozega--5-years' experience]. AB - In this paper, a 5-year-experience in arthroscopic meniscectomy at the Department of Orthopaedic Surgery, General Hospital Pozega, is presented. Patient data were obtained from medical documentation and physical examination after surgery and completed rehabilitation. We describe arthroscopic examination of the knee joint in our patients and finding of meniscal rupture, which ended with partial or subtotal meniscectomy. During the period since 1st January 1995 to December 31st 1999, at the Department of Orthopaedic Surgery in our hospital, 442 arthroscopies of the knee joints were performed. We found meniscal rupture in 235 patients (239 knees). In 232 knee joints partial or subtotal arthroscopic meniscectomy, and in seven knee joints arthrotomy and open meniscectomy according Smillie were done. In the last five years (1995-1999) the most common orthopaedic operation at the Department of Orthopaedic Surgery, General Hospital Pozega was arthroscopic meniscectomy (19% of orthopaedic operations). Complications (3.3%) of arthroscopic meniscectomy, which did not affect the good result of the treatment, are also presented. Authors discuss this and other arthroscopic procedures of treatment of meniscal rupture and specify their advantages and disadvantages. PMID- 11930757 TI - [Idiopathic eosinophilia with ascites (case report)]. AB - Hypereosinophilia can present a diagnostic problem if initial diagnostic procedures (medical history, physical examination and basic laboratory exams) fail to reveal the cause. Persistent finding of hypereosinophilia in such patients demands diagnostic intervention in order to rule out neoplastic and lymphoproliferative diseases, chronic inflammatory diseases of respiratory and gastrointestinal system and skin, and other inflammatory and postinflammatory conditions. If these disorders are ruled out, consideration should also be given to the diagnosis of primary hypereosinophilia, including idiopathic hypereosinophilic syndrome. The paper presents a patient with significant elevation of absolute and relative numbers of eosinophils in whom the only physical pathologic finding was eosinophilic ascites. Extensive diagnostic investigation did not prove secondary character of abnormalities, and since all pathologic findings receded during 7-month observation without treatment, the conclusion was that the disorder was benign. PMID- 11930756 TI - [Twin pregnancies in maternity wards in Cakovec, Sisak and Sibenik in 1998]. AB - This study shows the frequency of twin births in maternity wards where a pilot study of the use of child health cards was undertaken during 1998 (Cakovec, Sisak, Sibenik), the months in the year when twin births are more common, their characteristics in terms of the mother's age and parity, gestational age and the sex of the twins. In the three maternity wards mentioned, 3.582 children were born in 1998, of which 38 births were of twins, which is an average of 1.03%. There is no statistically significant difference between the three hospitals in terms of the frequency of twin births (p > 0.05). The most twins were born in May (23.6%). The average age of the mothers who gave birth to twins was 29.5 +/- 10.8 years. In terms of parity, twins were most often (in 55.3% cases) born to women who had already had 2 to 4 babies. In terms of the gestational age of twins, 20 pairs of twins (52.6%) were born at term, 18 pairs (47.4%) were born prematurely, 39 boys were born (51.3%), and 37 girls (48.7%), that is 13 pairs of boy twins, 13 pairs of girl-boy twins and 12 pairs of girls twins. The authors consider that the child health card, along with all its uses relating to the monitoring of the growth and development of children, may also serve in the monitoring of perinatal factors of twin pregnancies. PMID- 11930758 TI - [Less invasive laparoscopic cholecystectomy: initial experience and literature review]. AB - Laparoscopic cholecystectomy using the 5 and 10 mm caliber instrument is still a gold standard for surgical treatment of the gallstone disease. Today it is possible to perform this operation with a less invasive manner, using the fine caliber instruments, whose diameter is under 5 mm. According to published data, less invasive laparoscopic cholecystectomy does not have a higher complication rate, and other parameters are very similar or even better, specially the cosmetic outcome. In this article with small patient number we confirmed better cosmetic outcome, described different operation techniques and warned about the shortcomings. A greater number of patients and randomized study is indispensable for better technique analysis. PMID- 11930759 TI - [Hepatic portoenterostomy and primary liver transplantation in the treatment of biliary atresia]. AB - Results show that the use of sequential surgical treatment, employing Kasai portoenterostomy in infancy, followed by selective liver transplantation for children with progressive hepatic deterioration yields improved overall survival. All children with successful Kasai portoenterostomy procedures who do not require orthotopic liver transplantation are survivors. Using newer transplant techniques, the 5-year survival rate for children who receive transplants with a primary diagnosis of biliary atresia was 82%. This yields an overall survival rate of 86% in this entire study population. Limited donor availability and increased complications after liver transplantation in infants less than 1 year of age mitigate against the use of primary liver transplantation without prior portoenterostomy for infants with biliary atresia. At present, these two operative procedures should be used as sequential and complementary modes of treatment rather than as competitive procedures. When biliary atresia is not recognized in infancy and established cirrhosis has resulted, primary transplantation should be offered as the initial surgical treatment. PMID- 11930760 TI - [Occupational exposure to harmful materials in artistic activities]. AB - Artists' activities such as sculptures, paintings, graphic designs, photography, restoration, glassblowing, glass cutting and ceramics are described. The hazards the artists are exposed to during their professional activities with potential dangerous effects on their health are listed. Diseases which can develop as a consequence of exposure to specific noxious agents in such in environment are described. Preventive measures (personal, technical and medical) which should be applied in order to prevent the development of diseases related to these specific working conditions are stressed. PMID- 11930761 TI - [Dr. Hugo Gjankovic (Buzet, 18 September 1893--Dubrovnik, 14 May 1981)]. AB - Prof. dr. Hugo Gjankovic is among the most prominent Croatian doctors of this century. He has studied medicine in Austria and has been promoted in Wienna in 1920. First he worked unpaid in Ginecology Clinics in Wienna and then in 1922. he was accepted as assistant doctor at newly established Surgery Clinic of the Medical Faculty University of Zagreb. He worked hard to become respectable surgeon and university teacher. He was an effervescent member of the Croatian Medical Association--lecturing, publishing numerous papers and participating at many congresses in the country and abroad. His compulsory transfer, for political reasons, to the Department of Surgery of the State Hospital in Sarajevo during the Second World Was ment four years-long break in his academic career. Returned to afterwar Zagreb and to the Clinic as assistant professor he continued to improve the operation techniques, to establish the surgeons associations and to lecture for his colleagues, for the medical students and to the general public. He has published several textooks of surgery. Insulted by the measures taken by the Medical Faculty Authorities towards himself, he felt forced to leave Zagreb in 1953. In his 60th he has accepted the position of the Head of the Department of Surgery in the General Hospital in Dubrovnik. Next seven years he has invested in the through reconstruction of that hospital, particularly the Department of Surgery. He has proved helpful to his younger colleagues in their education and academic advancement. There is a lot to be learned from, not only his numerous papers and books, but from his biography, too. PMID- 11930762 TI - [Epidemiology of arterial hypertension in Croatia EH-UH 2000]. PMID- 11930763 TI - [Adequacy of hemodialysis at the threshold of the third millennium]. PMID- 11930764 TI - Management of chronic hepatitis C. AB - One purpose of the Clinical Practice Guideline column is to increase the awareness of the broad availability of existing guidelines and recommendations on various health topics. The hepatitis C virus (HCV) currently accounts for 20% to 40% of acute viral hepatitis, 60% to 80% of chronic hepatitis, and 20% to 30% of cirrhosis, end-stage liver disease, and liver cancer. Nearly four million Americans are currently infected with hepatitis C. Nurse practitioners should be aware of the recommendations regarding the diagnosis, management, and monitoring of the disease. This column summarizes the content of two NIH documents regarding the care of patients with hepatitis C: Management of Hepatitis C: NIH Consensus Statement (NIH, 1997) and Chronic Hepatitis C: Current Disease Management (NIDDK, 2000). PMID- 11930765 TI - Discovering resources--making global connections, international networking. AB - PURPOSE: To describe the key objectives, aims, activities and vision for the International Nurse Practitioner/Advanced Practice Nursing Network (INP/PNN). DATA SOURCES: Selected publications and documents, personal experience, and commentary. CONCLUSIONS: Advanced practice nursing and NP roles are prevalent in the United States and are now emerging in numerous countries throughout the world. There is a heightened interest among APNs and NPs internationally to share knowledge, expertise, and resources to enhance the presence of nursing in primary healthcare worldwide. The INP/APNN proposes to be an option for supporting the diversity of international networking in this field. IMPLICATIONS FOR PRACTICE: Nurse practitioner and APN roles are emerging globally. Globalization and global health issues impact all health care practitioners. Collaboration, partnering, and networking have the potential of enhancing and advancing practice for both experienced practitioners and countries that are initiating APN roles. PMID- 11930766 TI - Unintentional injuries in the young adult male. AB - PURPOSE: To review the primary causes of unintentional deaths in young adult males and suggest strategies for educating young men about reducing risks for accidental death. DATA SOURCES: Selected scientific literature and Internet sources. CONCLUSIONS: Unintentional deaths are the fifth leading cause of death in all age groups, after heart disease, cancer, stroke, and chronic obstructive pulmonary disease. Accidental death is the leading cause of death in males 1 to 44 years of age. Motor vehicle crashes, drowning, gunshot wounds, and occupational fatalities are among the leading injuries that contribute to the unanticipated death of the young adult male. IMPLICATIONS FOR PRACTICE: The ability to make sound decisions is a developmental process that is not achieved until later in adolescence or early adulthood. Nurse practitioners must assess the adolescent developmentally to provide appropriate guidance and assistance with behavior change. PMID- 11930768 TI - A review of antiphospholipid antibody syndrome. AB - PURPOSE: To review the pathophysiology, clinical presentation, and management options for antiphospholipid antibody syndrome (APS), a potentially life threatening coagulation disorder. DATA SOURCES: Selected scientific literature, consensus guidelines, and expert opinion. CONCLUSIONS: Clinical features that should alert the clinician to consider APS include recurrent fetal loss, arterial or venous thrombosis, thrombocytopenia and livedo reticularis. One should be suspicious of this diagnosis in a younger patient, one with an autoimmune disease, or family history of autoimmune disease. To confirm the diagnosis one needs both clinical and laboratory abnormalities. IMPLICATIONS FOR PRACTICE: The signs and symptoms of APS are varied and could be confused with many disorders. The primary care provider needs to be aware of this syndrome in order to include it in the differential diagnosis and appropriately recognize and refer the patient in a timely manner. PMID- 11930767 TI - Training of nurse practitioners and physician assistants to perform screening flexible sigmoidoscopy. AB - PURPOSE: To describe the colorectal cancer-screening program at Harvard Vanguard Medical Associates, a large multispecialty medical group, in which nurse practitioners (NPs) and physician assistants (PAs) perform screening flexible sigmoidoscopies. DATA SOURCES: Scientific literature, consensus statements and guidelines, and the protocol utilized to train NPs and PAs to perform flexible sigmoidoscopy. Data from 9,500 screening procedures are presented. CONCLUSIONS: In comparison with gastroenterologists, trained NP and PA endoscopists perform screening flexible sigmoidoscopy with similar accuracy and safety but at lower cost. IMPLICATIONS FOR PRACTICE: Screening flexible sigmoidoscopy performed by NPs and PAs may increase the availability and lower the cost of flexible sigmoidoscopy for colorectal cancer screening. PMID- 11930769 TI - Wait time and satisfaction with care and service at a nurse practitioner managed clinic. AB - PURPOSE: To examine the relationships among the various times patients wait for health care and patient satisfaction with nurse practitioner (NP) care and the service component of care in a NP developed and managed clinic. DATA SOURCES: Forty-seven patients (78% response rate) responded anonymously to a 15-item questionnaire that measured satisfaction with NP care and an additional 15 items that measured satisfaction with the service component of care. Actual clock times were recorded on a flow sheet that corresponded to the system of care at this clinic. CONCLUSIONS: Using Pearson Product-Moment Correlation, there were no statistically significant relationships among various wait times and the measures of satisfaction. IMPLICATIONS FOR PRACTICE: Patient satisfaction with NP care is an important indicator of quality of care. Although satisfaction was not related to wait times, NPs must be respectful of the amount of time that patients wait for health care. PMID- 11930770 TI - Men and family planning: what is their future role? AB - PURPOSE: To describe men's desired involvement in family planning and to determine the services desired by potential male clients. DATA SOURCES: Using a self-administered questionnaire, this study surveyed male partners of family planing clients and men attending sexually transmitted disease (STD) clinics at an urban health department. Perceived health concerns, contraceptive attitudes and practices, and desired involvement in family planning currently and in the future were targeted. CONCLUSIONS: Respondents reported desiring involvement in family planning decisions, although reported behaviors often conflicted with this desire. Routine physical examinations and receipt of health information were perceived to be important, while "male only" clinics were not. Respondents were willing to attend partners' family planning appointments if asked and were willing to help pay for the chosen contraceptive. Provision of vasectomy services was perceived as important. Prevention of cancer, STDs, and impotence were the three highest health concerns reported. IMPLICATIONS FOR PRACTICE: Men want to be partners in family planning and will access services if available. Current political and social policies are demanding more personal responsibility for the outcome of unintended pregnancies. In response to political and social demands, Title X family planning clinics are refocusing services to include men. PMID- 11930772 TI - Bringing up adultolescents. PMID- 11930771 TI - A crazy system. PMID- 11930773 TI - Deaf to good sense. PMID- 11930774 TI - A troubled mind. What it's really like to live with schizophrenia. PMID- 11930775 TI - Body and soul. Powerless prayer. PMID- 11930776 TI - Death of a liver donor. PMID- 11930777 TI - Cancer and diet. Colon vitamins. PMID- 11930778 TI - The madness of the elk. PMID- 11930779 TI - Environment. Floating pharmacy. PMID- 11930780 TI - Brain implants. Look, no hands! PMID- 11930781 TI - If you want honesty, break some rules. AB - We've all heard, or perhaps even told, the "organizational lie"; We're customer centric; everyone's performance is above average; we're the darling of our industry, coming up with one innovation after another. That last one was true of Advanced Cardiovascular Systems (ACS) in the past, but not when Ginger Graham took over as CEO. From that first moment in 1993, Graham chose to tell the truth about ACS's situation--that R&D was practically at war with product development, yields were down, and customers were disgruntled. And ever since, she's seen the benefits of exploding organizational lies. Truth telling is something that's hard to argue with but difficult to do. And, indeed, ACS instituted some radical practices to create its culture of honesty. Every senior manager was assigned a coach from the ranks who regularly solicited feedback from everyone, high and low, about the executive's performance. To get the truth, though, ACS executives learned that they had to offer it up themselves--the whole truth about the company's financial state, its problems, and its triumphs. When they did, they found that, in return, they could ask their employees for help in solving the problems, and passive complainers became active partners in the company's fortunes. ACS management spreads the word about the virtues of honesty through vivid stories of corporate history and quirky rituals. Every quarter, it holds companywide meetings in which the faults of top managers are examined--to keep them honest and tough enough to go on telling the truth. In fact, in the process of openly owning up to problems and jointly fixing them with employees, the entire company grew more powerful, nimble, and tough-minded, able to respond quickly to change, both internal and external. PMID- 11930782 TI - Wealth happens. AB - The economic world is full of patterns, and one of the most controversial is the distribution of wealth. You might expect the balance between rich and poor to vary widely from country to country. But back in 1897, Vilfredo Pareto discovered a pattern of wealth distribution that appears to be universal. Whenever you double the amount of wealth within a country, the number of people in each successively higher wealth bracket falls by a constant factor. The factor varies among countries, but the pattern remains essentially the same. From a mathematical standpoint, Pareto's distribution has stubbornly defied explanation. But recently, researchers were able to replicate the curve by applying the principles of network organization. They began with two simple assumptions. First, wealth accumulates either by transfers from person to person or through investment returns, positive or negative. Second, rich people invest more money than poor people. Starting with a hypothetical group of 1,000 people of equal wealth and abilities, the model always produces Pareto's wealth distribution no matter how the links in the network are organized or how the balance between interpersonal transactions and investment returns are set. The model also indicates that the degree of wealth concentration can be influenced. Increasing the number of links in the network or the total amount of money flowing through an economy tends to decrease wealth disparities; increasing investment returns or volatility tends to increase it. Replete with public policy implications, the model is only one example of how network analysis can reshape our understanding of complex economic and social systems, which may have less to do with the behavior of individual members than with impersonal and seemingly insignificant forces. PMID- 11930783 TI - Executive women and the myth of having it all. AB - When it comes to having a high-powered career and a family, the painful truth is that women in the United States don't "have it all." At midlife, in fact, at least a third of the country's high-achieving women--a category that includes high wage earners across a variety of professions--do not have children. For many, this wasn't a conscious choice: Indeed, most yearn for motherhood. So finds economist Sylvia Ann Hewlett, who recently fielded a nationwide survey to explore the professional and private lives of highly educated and high-earning women. Other findings are similarly disturbing. Many of these women who are raising children have suffered insurmountable career setbacks. In addition, today's young women will likely experience even starker trade-offs. In general, Hewlett's data show that, for too many women, the demands of ambitious careers, the asymmetries of male-female relationships, and the difficulties of conceiving later in life undermine the possibility of combining high-level work with family. By contrast, Hewlett's research reveals that high-achieving men continue to "have it all." Of the men she surveyed, 79% report wanting children, and 75% have them. Indeed, the more successful the man, the more likely he is to have a spouse and children. The opposite holds true for women, particularly the highest-achieving women in Hewlett's survey: At age 40, 49% of these ultra-achievers are childless, while 19% of their male peers are. The facts and figures are bleak but, the author contends, could be liberating if they spur action. Hewlett urges lawmakers and corporations to establish policies that support working parents. But recognizing that changes won't happen overnight, she exhorts young women to be more deliberate about their career and family choices. PMID- 11930784 TI - Reawakening your passion for work. AB - All of us struggle from time to time with the question of personal meaning: "Am I living the way I want to live?" For millions of people, the attacks of September 11 put the issue front and center, but most of us periodically take stock of our lives under far less dramatic circumstances. This type of questioning is healthy; business leaders need to go through it every few years to replenish their energy, creativity, and commitment--and their passion for work. In this article, the authors describe the signals that it's time to reevaluate your choices and illuminate strategies for responding to those signals. Such wake-up calls come in various forms. Some people feel trapped or bored and may realize that they have adjusted to the frustrations of their work to such an extent that they barely recognize themselves. For others, the signal comes when they are faced with an ethical challenge or suddenly discover their true calling. Once you have realized that it's time to take stock of your life, there are strategies to help you consider where you are, where you're headed, and where you want to be. Many people find that calling a time-out--either in the form of an intense, soul searching exercise or a break from corporate life--is the best way to reconnect with their dreams. Other strategies include working with a coach, participating in an executive development program, scheduling regular time for self-reflection, and making small changes so that your work better reflects your values. People no longer expect their leaders to have all the answers, but they do expect them to try to keep their own passion alive and to support employees through that process. PMID- 11930785 TI - Saving your rookie managers from themselves. AB - Most organizations promote employees into managerial positions based on their technical competence. But very often, that kind of competence does not translate into good managerial performance. Many rookie managers fail to grasp how their roles have changed: that their jobs are no longer about personal achievement but about enabling others to achieve, that sometimes driving the bus means taking a backseat, and that building a team is often more important than cutting a deal. Even the best employees have trouble adjusting to these new realities, and that trouble can be exacerbated by the normal insecurities that may make rookie managers hesitant to ask for help. The dynamic unfolds something like this: As rookie managers internalize their stress, their focus, too, becomes increasingly internal. They become insecure and self-focused and cannot properly support their teams. Invariably, trust breaks down, staff members become alienated, and productivity suffers. In this article, coach and management consultant Carol Walker, who works primarily with rookie managers and their supervisors, addresses the five problem areas that rookie managers typically face: delegating, getting support from senior staffers, projecting confidence, thinking strategically, and giving feedback. You may think these elements sound like Management 101, and you'd be right, Walker writes. But these basic elements are also what trip up most managers in the early stages of their careers (and even, she admits, throughout their careers). The bosses of rookie managers have a responsibility to anticipate and address these problems; not doing so will hurt the rookie, the boss, and the company overall. PMID- 11930786 TI - "The expert witness: death of an oxymoron". PMID- 11930787 TI - The Lloyd Roberts Lecture. Immunopathology: past, present and future. PMID- 11930788 TI - Hamilton Bailey--a surgical tale of triumph and disaster. PMID- 11930789 TI - The Brighton and Sussex Medicochirurgical Society. PMID- 11930790 TI - York Medical Society. PMID- 11930791 TI - The Liverpool Medical Institution: a brief historical summary. PMID- 11930792 TI - The Medical Society of London. PMID- 11930793 TI - Modern cosmology--God and life. PMID- 11930794 TI - Joint operations: articulation in the family division. PMID- 11930795 TI - Fothergillian Lecture. Russian medical education: then and now. PMID- 11930796 TI - Lettsomian Lecture. Hansen's warning. PMID- 11930798 TI - The physical, mental and moral health of prisoners and a policy of improvement. PMID- 11930797 TI - A short history of the Jenner building. PMID- 11930799 TI - Benjamin Franklin and the Craven Street bones. PMID- 11930800 TI - [Disorders of antioxidant defence and their correction in experimental di isopropyl-phenyl-isocyanate intoxication]. AB - Using preventively natural and artificial antioxidants in experiments on white rats enabled to assess role of lipids peroxidation in mechanisms of acute inhalation toxicity caused by 2,6-diisopropylphenylisocyanate. Correlation and regression analysis revealed differences in mechanisms underlying antioxidants effects. PMID- 11930801 TI - [Aspects of exposure to methyl-methacrylate of varying intensity in an organism and on population levels]. AB - Studies of mortality among 3588 workers who contacted variable doses of methylmetacrilate (MMA) revealed that higher intensity of the exposure led to increased risk of death with lung and colon cancer. Immune state studies in 157 workers proved significant immune disorders due to MMA levels several times exceeding the MAC. Experiments evaluating immune functions under exposure to MMA levels equal to or under the MAC demonstrated mostly adaptational changes. PMID- 11930802 TI - [Chloroprene as possible industrial carcinogen]. AB - The authors conducted 2 independent cohort studies on epidemiology of malignancies among workers contacting chloroprene. The first study covered morbidity and mortality with malignancies among workers engaged into polychloroprene resins production in Erevan; the second study--mortality with malignancies among Moscow shoe makers who contacted polychloroprene glues and latex. The findings are reliably higher risk of morbidity and mortality with hepatic cancer, dose--effect dependence between chloroprene exposure and occurrence of and mortality with hepatic cancer. PMID- 11930803 TI - [Some approaches to informatics in occupational therapy service]. AB - The authors present results obtained by local occupational therapy centers and concerning tests of new account forms designed by Occupational Medicine Research Institute with RAMSc for informational background of occupational therapy service. PMID- 11930804 TI - [Management forms to improve early diagnosis and prophylaxis of occupational diseases in workers of major chemical enterprise]. AB - Cooperation of occupational therapists with local authorities and with Sanitary and epidemiologic supervision establishments results in definite positive changes in diagnosis and primary prophylaxis of occupational diseases, improves quality of medical examinations. Prevention of occupational diseases results from contract system in hiring for hazardous work conditions. The job contract should invariably include a medical part that definitely settle rights and duties associated with health preservation. PMID- 11930805 TI - [Methods detecting boron compounds in hygienic evaluation of environment]. AB - Highly sensitive, selective express-methods are designed to determine boron compounds in atmosphere and in the air of workplace, in river and sea waters, in biologic materials. The methods are put into practice on enterprises, could be useful for hygienic research. PMID- 11930806 TI - [Current problems of occupational health in polyurethane production]. AB - Prospective production of polyurethane is not safe for humans. This production is characterized by possible workers' exposure to chemical hazards mixture. The work safety includes cautionary and current sanitary supervision, implantation of sanitary recommendations concerning individual protective means also. PMID- 11930807 TI - [Chlorophenols: intermediate by-products of 2,4-dichlorophenoxyacetic acid synthesis. Toxicity and hazard]. PMID- 11930808 TI - [Length of employment and dosage in the evaluation of risk factors for chronic bronchitis among workers in contact with respiratory tract chemical irritants]. PMID- 11930809 TI - [International scientific and practical conference "Social and occupational problems in the assessment of health status and health services under contemporary conditions]. PMID- 11930810 TI - [Functional cardiovascular status in patients with high blood pressure and coronary heart disease in a remote period of uranium intoxication]]. AB - High blood pressure and coronary heart disease in far-off period of chronic uranium intoxication result in re-modelling of left ventricle with predominant concentric re-modelling. Lower myocardial mass of left ventricle and more marked changes in diastolic function occurred in the main group if compared to the reference one having no contact with occupational hazards. PMID- 11930811 TI - Effective and culturally acceptable water storage in Zimbabwe: maintaining the quality of water abstracted from upgraded family wells. AB - Because domestic water can be a vehicle of disease transmission in the home, there is a need for intervention. In Zimbabwe. 60 rural households obtaining water from shallow wells were selected for a field study. A water urn was designed, pretested, and field-tested. Thirty households designated as the case group were given two water urns each to substitute for traditional water storage containers (paint containers, oil containers, etc.). The remaining 30 households served as a control group. Samples were collected twice, at two-week intervals, from the water supply source (upgraded family wells) and storage containers (water urn or traditional containers) of each household (228 samples). Total coliform bacteria and fecal coliform bacteria were enumerated with the membrane filtration technique. A pretest of the water urn design showed a decline in turbidity that corresponded with a decline in bacterial counts. Wells supplying the case households had higher bacterial counts than those supplying the control households, but bacterial loads in the water urns were significantly lower than those observed in the corresponding supply wells (paired t-test: t = 3.97, df = 55, p < .01). Bacterial loads in traditional containers were similar to those observed in the supply wells (paired t-test; t = 0.2, df = 57, p > .05). The case group eagerly substituted water urns for traditional containers. The use of water urns was found to prevent or to reduce further contamination of well water after collection. PMID- 11930812 TI - Glimpses into NEHA's 2002 Nationwide Salary & Benefits Survey of Local/County Environmental Health Professionals. PMID- 11930813 TI - Proper operation, maintenance, and servicing of aerobic wastewater treatment systems. AB - Aerobic waste treatment systems are a safe, simple, and a cost-effective method of treating waste on site. With a small amount of care in the operation and maintenance of the system, these units will provide extremely high-quality effluent and fulfill an important need where conventional systems are not effective. PMID- 11930815 TI - Professional scientific presentations. PMID- 11930814 TI - The legal powers of health agencies--recent developments. PMID- 11930816 TI - Childhood lead poisoning investigations: evaluating a portable instrument for testing soil lead. AB - The Childhood Lead Poisoning Prevention Branch of the California Department of Health Services evaluated a portable X-ray fluorescence (XRF) instrument for use as a soil lead-testing tool during environmental investigations of lead-poisoned children's homes. A Niton XRF was used to test soil at 119 sampling locations in the yards of 11 San Francisco Bay Area houses. Niton XRF readings were highly correlated with laboratory results and met the study criteria for an acceptable screening method. The data suggest that the most health-protective and time efficient approach to testing for soil lead above regulatory levels is to take either surface readings or readings of a test cup of soil prepared by grinding with a mortar and pestle. The advantage of the test cup method is that the test cup with soil may be submitted to a laboratory for confirmatory analysis. PMID- 11930818 TI - Chest pain centers--Part 2. The strategy of the chest pain center. PMID- 11930817 TI - Chest pain centers--Part 1. Chest pain centers: past, present and future. PMID- 11930819 TI - Chest pain centers--Part 3. Evaluation in the hospital ED or chest pain center (CPC). PMID- 11930820 TI - Chest pain centers--Part 4. Executive summary: issues with APC's and observation services. PMID- 11930825 TI - Drug susceptibility testing of Malassezia furfur strains to antifungal agents. AB - The minimal inhibitory concentration of the three most commonly used antifungal agents--ketoconazole, fluconazole and intraconazole against 28 strains of Malassezia furfur, was studied. The strains were recovered from patients with pityriasis versicolor, pityriasis simplex capitis and dermatitis seborrhoides and were cultured on Dixon's agar. Diagnostic sensitivity test agar was used to determine the minimal inhibitory concentrations of the antifungal agents. The established minimal inhibitory concentration of ketoconazole against Malassezia furfur was 0.02 mg/l, the minimal inhibitory concentration of intraconazole--0.05 mg/l and that of fluconazole--0.09 mg/l. The results of the study are consistent with the data reported by other European authors in their studies. On the basis of the accumulated data it is concluded that the tested antifungal agents are effective in the treatment of infections caused by Malassezia furfur but ketoconazole is superior to intraconazole and fluconazole in the treatment of these infections. This is the first study on the drug susceptibility of Malassezia furfur in Bulgaria. PMID- 11930826 TI - In situ hybridization determination of the heterophagocytic origin of type 2B prostate epithelial lipochrome pigment granules: histochemical and ultrastructural correlates. AB - Morphologically, polymorphic prostatic lipochrome pigment has been classified and subclassified in the last few years. Type 2B lipochrome pigment granules (LPGs) are frequently found in prostatic epithelium in patients who had died of AIDS. Intensive apoptosis is observed in the same epithelium which lends support to the hypothesis of heterophagocytic (apoptotic) origin of type 2B pigment granules. Detection of nuclear chromatin material is necessary for the differentiation of an autophagosomal from a heterophagosomal structure in the cellular cytoplasm. OBJECT OF THE STUDY: Application of in situ hybridization (ISH) for elucidating the origin of subtype 2B LPGs in the prostate epithelial cells of patients who had died of AIDS. METHODS: ISH was used on routine necropsic prostate epithelial samples from three patients who had died of AIDS. A DNA probe raised against total human DNA was employed. RESULTS: Multiple hybridization signals were detected in type 2B LPGs which shows the presence of nuclear material in those structures. The chromatin material localized to the periphery of pigment granules. CONCLUSION: Type 2B LPGs have a heterophagocytic origin and represent phagocytosed apoptotic bodies in the phase of phagolysosomal degradation. They can be used as a morphologic tissue marker of intensive epithelial apoptosis. PMID- 11930827 TI - Stimulation electromyography as a method of intraoperative identification of the recurrent laryngeal nerve in thyroid surgery. AB - INTRODUCTION: The iatrogenic injury of the recurrent laryngeal nerve during thyroid surgery still represents a major problem because of the great physiologic and psychologic trauma to the patients. The incidence of this complication ranges between 1% and 15% depending on the thyroid pathology and type of surgery. AIM: To evaluate the potential of stimulation electromyography as a method of intraoperative prophylaxis of iatrogenic injuries of the recurrent laryngeal nerve. MATERIAL AND METHODS: Intraoperative monitoring of the recurrent laryngeal nerve based on stimulation electromyography, which was introduced in the Clinics of Surgery and Otorhinolaryngology at the Higher Medical Institute in Plovdiv, was applied in thyroid surgery using Neurosign 100 (Magstim Company Ltd., Wales, UK) apparatus. The study involved 5 experimental dogs and 73 recurrent laryngeal nerves in 47 patients treated in the clinics from November 1, 1999 to February 15, 2000. RESULTS: In all experimental animals and patients the electrical stimulation was successful triggering an "M" response of the laryngeal muscles (the vocal muscle). A stable contact was achieved between the recording (needle) electrodes and the vocal folds. None of the patients had signs of laryngeal palsy or complaints other from the usual in endotracheal intubation. CONCLUSION: The results suggest that the intraoperative stimulation electromyography of the recurrent laryngeal nerve is a safe and effective method for evaluation of the nerve integrity during and at the end of the operation. PMID- 11930828 TI - Discriminant analysis of minor physical anomalies as dysontogenetic markers of schizophrenia. AB - AIM: The aim of the study was to evaluate the predictive value of minor physical anomalies in the binomial model schizophrenic patient-mentally healthy subject as an index of developmental origin of schizophrenia. MATERIAL AND METHODS: Sixty eight schizophrenic patients (38 males and 30 females) and 69 mentally healthy subjects (36 males and 33 females) of Bulgarian ancestry were examined for minor physical anomalies using the Waldrop Physical Anomaly Scale. The predictive value of minor physical anomalies for schizophrenia was determined by discriminant analysis. RESULTS: The two-group discriminant analysis distinguishes well the schizophrenic patients from the controls with 6 predictive variables. The variables with significant contribution to prediction of the patient-control status are fine electric hair (p < 0.001), gap between I and II toes (p < 0.001), epicanthus (p < 0.001), high/steepled palate (p < 0.001), tongue with rough/smooth spots (p < 0.001), III toe > or = II toe (p < 0.001). The model overall correct classification is 78.63%, predicting slightly better the schizophrenics than the controls (79.7% vs. 77.4%). CONCLUSIONS: The constellation of variables entered into the discriminant analysis model confirms the hypothesis that at least in one subpopulation of schizophrenic patients the minor physical anomalies reflect early developmental processes determining occurrence of the disease in later periods of life. PMID- 11930829 TI - A factor analysis of dyspnea indexes and lung function parameters in patients with chronic obstructive pulmonary disease. AB - The purpose of the present study was to investigate the correlation between dyspnea ratings and a large group of lung function parameters, and extract those factors that best reflect the functional profile of patients with COPD using factor analysis. Ninety nine patients with COPD in stable clinical condition (age 60 +/- 8 years, ATS score = 2.5 +/- 0.9, FEV1% pred. = 33 +/- 13%) were included in the study. The factor analysis of the results yielded 5 factors which accounted for 80.1% of the total variance of the changes. The highest coefficients found between the factors and the original group of variables after Varimax rotation are given in the following table: Factor 1: Oxygen-cost diagram: 0.92; ATS dyspnea score: -0.80; TL,CO/VA: 0.78; Factor 2: FEV1% pred.: 0.87; FEV1/VC%: 0.86; FEV1L: 0.79; Factor 3: MIF50% pred.: 0.85; FIV1% pred.: 0.76; PImax: 0.67; Factor 4: PaCO2: -0.81; SaO2: 0.77; Mean pulmonary arterial pressure: -0.67 Factor 5: Age: 0.88; Six minutes walk distance: -0.72 The factor analysis showed that the functional profile of COPD patients has several dimensions. Therefore, in order to have COPD comprehensively evaluated, assessment of dyspnea and the respective set of lung function parameters (exercise capacity, forced inspiration and pulmonary hemodynamics), should be included in the battery of tests, besides the conventional tests. PMID- 11930830 TI - Optimisation of the technique of computed tomographic imaging of paranasal sinuses. AB - BACKGROUND: Recently, computed tomography has won recognition as a standard method for confirming opacifications in the sinuses because of its high resolution, absence of superimposition in visualizing objects, and the extremely accurate density differentiation of the tissues. With the advent of functional endonasal surgery this technique acquired especial significance in the preoperative imaging of sinuses with inflammatory diseases and the postoperative follow-up of the results. OBJECTIVE: The aim of the present study was to optimize the imaging technique of the paranasal sinuses. MATERIAL AND METHODS: Fifty patients with inflammatory disorders of the paranasal sinuses were examined using Sytec 3000 scanner. We developed a special technique for coronal and axial scanning. RESULTS: The bone and soft tissues were very clearly delineated in every slice. We used lower exposure factor which reduced the radiation dose by 40%. CONCLUSIONS: The technique we developed provides a standard for imaging the sinuses; it allows accurate preoperative diagnosis and postoperative follow-up of the results of the surgery in this region. PMID- 11930831 TI - Study of the titer of common antibodies against superficial antigens of Pityrosporum ovale in dermatitis seborrhoides. AB - Pityrosporum ovale is the yeast form of the dimorphic lipophilic fungus Malassezia furfur which is part of the normal skin microflora in adults. It is suggested that Pityrosporum ovale has a primary role in the pathogenesis of Dermatitis seborrhoides. The aim of the present study was to determine the titer of the common antibodies against superficial antigens of Pityrosporum ovale in the serum of Dermatitis seborrhoides patients and compare the results with the findings of examinations of serum antibodies in healthy subjects and patients with Pityriasis simplex capitis, Pityriasis versicolor, Pityrosporum folliculitis. Indirect immunofluorescent technique was used in the study. The Pityrosporum ovale antigen was obtained using own method. The results show that Dermatitis seborrhoides patients have an antibody titer of 1:640 and above, which is a higher titer than the antibody titers of healthy subjects and patients with other forms of the infection. The study results suggest that Pityrosporum ovale yeast might be implicated in causing a humoral immune response, which is important for the developing inflammatory reactions in Dermatitis seborrhoides. The results help to elucidate the pathogenic mechanisms and supplement the therapy strategy of the disorder. The authors are the first in Bulgaria to study the immune response to Pityrosporum ovale in healthy subjects and patients suffering from different forms of the infection. PMID- 11930832 TI - One-stage reconstruction of the eye socket with vascularized temporalis muscle flap in orbital exenteration. AB - AIM: Two cases of one-stage reconstruction of exenterated orbit with vascularized temporalis muscle flap in oncological patients are presented. The anatomical characteristics of the temporalis muscle flap and the choice of one-stage or delayed reconstruction of the exenterated orbit are discussed. METHODS: Two patients were operated for malignancy recurrence during the period May-November 2000. One patient suffered recurrent squamous cell carcinoma in the left nasolabial region that infiltrated the lower eyelid and periorbit. The other patient had recurrence of previously operated primary rhabdomyosarcoma of the orbit. Both patients underwent orbital exenteration. In the first patient it supplemented the tumor excision and partial maxillary excision resulting in large orbitomaxillary defect communicating with the nasal meatuses. RESULTS: In both patients good filling effect and isolation of the orbit from the neighbouring nasal meatuses and cranial cavity were obtained using vascularized temporalis muscle flap. CONCLUSION: The vascularized muscle flap appears a reliable technique and method of choice in one-stage as well as in delayed reconstruction in orbital exenteration. PMID- 11930833 TI - Primary localized amyloidosis of the urinary bladder. A case report. AB - We present case 96 (the first one in Bulgaria) of primary localized amyloidosis of the urinary bladder in a 72-year-old woman with episodes of painless hematuria. The amyloid tumor is situated on the anterior wall of the bladder--a yellowish prominence 15 mm in diameter with superficial erosion that is surrounded by a reddish ring and bullous edema of the mucosa. Rough and regularly pink perivascular deposits, thickened vessel walls and focal lymphoplasmocyte infiltrates are present in the submucosa. After preliminary treatment with KHMnO4, AL-amyloid is detected in polarized light by Congorot staining and in ultraviolet light using thioflavin. Immunoglobulins and C3 deposits are not found. On electron microscopy deposits of amyloid fibrils are seen in abundance close to collagen bundles. Diseases that may be associated with systemic amyloidosis have been clinically excluded. Nodular amyloidosis resulting from a local immune dyscrasia is accepted in the case. The patient is in good health 11 months after the first biopsy. A literature review on this rare form of localized amyloidosis has been made. Generally, the disease has a favorable prognosis and non-active behavior is recommended. Intravesical instillations of dimethyl sulfoxide, sectional resection or plastic surgery that preserve the urinary tract are used in the rare case of progressive course. PMID- 11930834 TI - Clinico-electrophysiological correlates in patients with relapsing-remitting multiple sclerosis. AB - INTRODUCTION: The degree of neurological deficit was evaluated in patients with multiple sclerosis (MS) relapse by the Expanded Disability Status Scale (EDSS), transcranial magnetic stimulation (TMS) and visual evoked potentials (VEPs). Clinical and electrophysiological changes before and after treatment were compared in an attempt to find significant associations between the indicators of disease activity. MATERIAL AND METHODS: In all the patients (n = 35, mean age 32.05 +/- 2.67 years) EDSS rating was assessed prior to and 2 months after the beginning of treatment with methylprednisolone (MP)--13 patients, and with MP in combination with cyclophosphamide (CPH)--22 patients. Transcranial magnetic stimulation with measuring central latency time (CLT) was performed in 20 patients (mean age 30.85 +/- 8.01 years) before and at the end of the 2nd month after treatment with MP independently (9 patients) and MP combined with CPH (11 patients). In another 15 patients with a mean age of 33.67 +/- 7.45 years VEPs are examined (P100 latency) before and on the 2nd month after treatment with MP (4 patients) and MP combined with CPH (11 patients). RESULTS: CLT (P < 0.001) and P100 (P < 0.001) were significantly shortened after treatment in comparison with the baseline values. A significant correlation (P < 0.05) between CLT and the pyramid subscale was found. CONCLUSION: The results suggests that CTL and P100 reflect the effect of treatment on the intensity of the pathological process, the CLT correlating with the severity of the pyramid deficit according to EDSS rating in the course of the treatment. PMID- 11930835 TI - A case of undifferentiated (embryonal) sarcoma of the liver: fine needle aspiration cytology diagnosis by one cell type. AB - We present the first case in Bulgaria of cytologically verified undifferentiated (embryonal) sarcoma of the liver. A 10-year-old girl was admitted to the hospital with abdominal pain, extreme hepatomegaly, one-month fever, anemia and highly elevated erythrocyte sedimentation rate (95 mm). A large formation was found in the right liver lobe on ultrasound examination, which was polycystic on CT scan. Neither bone marrow abnormalities nor elevation of serum alpha-fetoprotein was found. In the scanty specimen obtained by fine-needle aspiration biopsy under ultrasound control a characteristic prevalence of polygonal cells within mesenchyme tissue was found. The cells had eccentric hyperchromic nuclei which were pushed to the periphery by large clusters of PAS-positive, diastase resistant granules. Vimentin and alpha 1-antitrypsyn were expressed immunohistochemically. The cells are pathognomonic of undifferentiated (embryonal) sarcoma of the liver. After preoperative chemotherapy with ifosfamide, vincristine and actinomycin D (CEVAIE, CWS--96) resection of the tumor including IV, V, VI and VII liver segments was performed. A 30 x 30-cm soft gray-whitish tumor with fine capsule was found. The cytological diagnosis was histologically verified. The patient had a mechanical subileus episode after the 6th course of chemotherapy. One year after the surgery she is in good condition. The histogenesis of this rare tumor is discussed regarding its unique cytology, immunohistochemical constellation and ultrastructure. A literature review is presented. The case indicates that the unique cell appearance in embryonal sarcoma of the liver provides a correct diagnosis even in a scanty specimen. PMID- 11930836 TI - [Human tooth tissue as dental restorative material]. AB - With the recent development of copy-milling systems for porcelain, it has now become possible to construct fitting restorations from natural human tooth substance. This preliminary study aimed to find out whether it is possible to produce a well fitting Class II inlay from an extracted tooth using a copy milling machine, so that the inlay produced will have enamel over its whole outer surface and dentine on its inner surface. Two pairs of matching sound extracted permanent molars were used. The molars were matched for mesio-distal size of the tooth crown and the convexity of the approximal surfaces. One tooth of each pair was assigned to be the 'donor' tooth, the other tooth being the 'host'. MO inlay preparations were made in the 'host' teeth. These were restored with the milled 'natural inlays' produced from the 'donor' teeth using the CELAY system. The restored 'host' teeth were sliced and examined using light-microscopy. All specimens showed a good fit and the desired location of enamel and dentine. The milling accuracy was assessed in a second experiment. This experiment showed similar marginal gaps for 'natural inlays' as for ceramic inlays. This study has shown that by using the CELAY milling machine it is possible to 'recycle' extracted teeth for the production of accurate restorations. PMID- 11930837 TI - [Early orthodontic treatment?]. AB - Today orthodontists in the United States tend to treat much earlier. During an early treatment phase, removable appliances are used to correct the gross malocclusion. After a subsequent retention-observation phase the remaining orthodontic anomalies are corrected using fixed appliances. In this review article the advantages and disadvantages of early orthodontic treatment are evaluated. PMID- 11930838 TI - [Syndromes 7. Hypertelorism (Greig syndrome)]. AB - Hypertelorism is an increased distance between the orbits. Most evident symptom is the increased intercanthal distance. It can be caused by congenital deformities (facial clefts, encephaloceles, enlarged ethmoids) and by trauma. Three degrees are differentiated. Grade one and two can be corrected with a small risk for complications. If there is vertical orbital dystopia it has to be corrected by a transcranial procedure, as is necessary in grade 3. PMID- 11930839 TI - [Front bridge damaged by accident?]. PMID- 11930840 TI - [Melanoma]. PMID- 11930841 TI - [Abscesses caused by deciduous teeth]. AB - Abscesses in children, caused by deciduous teeth, ar not so frequently as seen in the permanent dentition. In most cases caries or an odontogenic infection of the associated deciduous tooth have already been treated by the dentist or oral surgeon at an earlier stage, i.e. before the infection exacerbates to an abscess. At the department of Oral and Maxillofacial Surgery of the Academic Hospital Nijmegen, between April 1997 and April 1998 20 children have been treated on a total of 1,693 patients with an odontogenic infection. Of this group, nine young children had an abscess caused by a deciduous tooth. In most cases the mandibular deciduous molars were involved. In one case an osteomyelitis of the mandible was diagnosed. All abscesses and infections could be treated adequately. The use of antibiotics is of paramount importance in the treatment of these abscesses in children. PMID- 11930842 TI - [Dentistry: the twentieth century and the future. Opinions of Dutch dentistry professors]. AB - The main objective of this study was to investigate the opinions of Dutch dentist professors about the main developments in dental science and Dutch dental care in the period 1900-2000 and their expectations for the coming 25 years. In the Dentist Directory of the Dutch Dental Association 51 professors were found of which 23 were still active; 45 were interviewed. The answers were categorized into 5 groups. The majority of the respondents (n = 31) mentioned as the most important development in dental science in the past century the gathered knowledge of the preventive effects of fluoride and the wide application of it in practice. The foundation of the dental health insurance in the Dutch National Health system was seen as the most significant development in Dutch dental care in this period (n = x32). More than half of the group (n = 26) was worried about the future demand of dentists and about the coming problems of supply and demand of dental care. It is concluded that the results of this study seem to be an accurate representation of the opinions of Dutch dentist professors on the main developments in dental science and Dutch dental care in the period 1900-2000. Respondents were pleased with the dental advances in the past century but they felt uneasy for the coming 25 years, especially concerning the issue of the availability of dental care for the majority of the Dutch population. PMID- 11930843 TI - [The changing role of the dental hygienist]. AB - This article describes the role of the dental hygienist in the Netherlands from the start of the training of dental hygienists in 1968. With the new Individual Health Care Professionals Bill from 1997 the dental hygienists are entitled to perform more tasks than in the past. This article describes the viewpoint of the professional dental hygienist organization on the future role of the dental hygienist. Cooperation with the other members of the dental team and free entry to the dental hygienist are considered important conditions. The creation of appropriate conditions by government and insurance companies is a matter of major concern. PMID- 11930845 TI - [The Ivory Cross: past, present and future. Promotion of oral health in the Netherlands by a national information organization]. AB - In this article a description is given of the role of Dutch National Association for the Promotion of Oral Hygiene (Ivoren Kruis), the Dutch organisation of oral health education. In this review attention is paid to the past and to the recent working method of the association by which it tries to realise the improvement of the dental health on the population and individual level. Special attention is paid to the evaluation of all sort of activities and programmes on oral health education. The article concludes with a short view on the future of the organisation. PMID- 11930847 TI - [A matter of taste]. PMID- 11930846 TI - [Syndromes 16. Osteogenesis imperfecta]. AB - Osteogenesis imperfecta consists of a group of hereditary connective tissue diseases with fragility of the bone as a general feature. Frequently occurring other characteristics are blue sclerae, opalescent teeth, hearing loss, disorders of the skeleton, and hyperextensibility of the joints. Obliteration and the unusual shape of the pulp chambers may interfere with dental treatment. PMID- 11930848 TI - [A patient with thrombocytopenia: what do I do?]. PMID- 11930849 TI - [Stomach cancer]. PMID- 11930850 TI - Infectious diseases: filariasis, Malayan filariasis, loiasis (ioa ioa), Onchocerciasis (river blindness). PMID- 11930851 TI - Urinary tract infection. AB - The University of Michigan Health System Urinary Tract Infection CPG provides a summary of evidence-based recommendations on the treatment of UTIs in a format that can easily applied in practice. The authors include a statement of the importance of clinical judgement in the application of the recommendations and the recognition that other treatment approaches may be appropriate. The panel of authors includes physicians of various specialties, an ambulatory care nurse, and a medical education specialist. While the specialties of general and family medicine, OB/GYN, and infectious medicine are represented, there is no urologist or nephrologist listed on the panel. The CPG includes a brief annotated bibliography, summarizing six of the articles considered in the guideline's development. There is no further extended list of references included in the content. The CPG is limited to the treatment of adult women, although there is no specific age cited at which the guideline would begin to apply. It provides the reader with ideas for not only managing women who present to the clinic with symptoms consistent with a UTI, but also ideas for not only managing women who patient self-treatment strategies to minimize the cost associated with recurrent infections. Health care professionals interested in using the CPG in practice are encouraged to obtain and review a full-copy. It is available on the World Wide Web ate cme.medumich.edu.pdf/guideline/UTI.pdf. PMID- 11930852 TI - Rheumatoid arthritis: a primary care approach. AB - PURPOSE: To provide primary care providers with guidelines on the evaluation, diagnosis and management of patients with rheumatoid arthritis (RA). DATA SOURCES: Research-based articles in the medical literature, review articles, and clinical practice is a chronic inflammatory disease that affects mainly synovial joints. Diagnosis is based primarily on clinical examination. Clinical criteria developed by the American Rheumatism Association can help the primary care provider recognize this potentially devastating disease and facilitate early referral to a rheumatologist for treatment. IMPLICATIONS FOR PRACTICE: Patients with RA should be treated early and aggressively. Laboratory tests are not essential to confirm the diagnosis of RA. The clinical usefulness of the rheumatoid factor could be increased by restricting the test to patients who have a high probability of RA based on clinical symptoms. PMID- 11930853 TI - Primary care assessment and management of sleep disorders. AB - PURPOSE: To review the functions of sleep and describe an appropriate sleep assessment for the identification of sleep disorders for nurse practitioners (NP) in primary care. Guidelines for management and referral are included. DATA SOURCES: Selected and the author's experience. CONCLUSIONS: The incidence of sleep deprivation in the United States is generally unacknowledge by the public as well as health care professional. Unfortunately, the castastrophic and debilitating consequences that result from alterations in sleep are usually noted after significant physical and emotional damage have occurred. IMPLICATIONS FOR PRACTICE: An assessment of sleep hygiene and the physical, emotional, or lifestyle changes that alter sleep patterns should be a part of the NP's routine encounter with patients. Referral is indicated for suspicion of obstructive sleep apnea, narcolepsy, restless leg syndrome, complex motor activity, or if the condition fails to respond to therapeutic management. PMID- 11930854 TI - College student's attidudes and beliefs about the consequences of smoking: development and normative scores of a new scale. AB - PURPOSE: To develop an instrument that could be used to assist young adults to determine their perceived consequences about cigarette smoking. The new instrument consisted of 27 items measuring attitudes about smoking selected from the literature and to a convenience sample of 172 undergraduate college students. Psychometric assessment using an exploratory factor analysis revealed four factors (subscales) that explained 48% of the variance. These were labeled attitudes and beliefs about smoking related to emotional benefits, health hazards, self-confidence, and body image. CONCLUSIONS: Statistically significant differences between mean scores of smokers and nonsmokers were found on attitudes and beliefs about the benefits of smoking related to emotional benefits, self confidence, and body image; smokers' answers indicated that they perceived these as positive consequences of their smoking behavior. IMPLICATIONS FOR PRACTICE: The newly constructed instrument may be a useful assessment of college students' reasons for smoking. This approach offers new hope for successful cessation counseling and for smoking prevention programs. Currently antismoking methods emphasize the health hazards of cigarette smoking and have not been sufficient in reducing the rate of smoking in the young adult population. Some young people may use smoking as strategy for dealing with stressful situations, weight control, and lack of self-confidence. Thus, smoking cessation programs should also include strategies to use in place of smoking during periods when stress and lack of self confidence are high. PMID- 11930855 TI - Ending a nurse practitioner-patient relationship: uncovering patients' perceptions. AB - DATA SOURCES: Seventy-nine patients assigned to the care of a nurse practitioner (NP) were interviewed to explore reactions to ending a yearlong therapeutic relationship at the conclusion of a clinical trial. Three researchers identified, reviewed and CONCLUSIONS: Of the total 79 patients, 22 (28%) spontaneously discussed perceptions and feelings about the termination of their relationship with the NP, Qualitative analysis of their statements identified future concerns about continuity of care and emotional themes ranging from gratitude, regret, and anxiety to grief. IMPLICATIONS FOR PRACTICE: Changes in health care coverage often result in abrupt termination of patient-provider relationships. The involuntary termination of a patient-provider relationship may have significant negative consequences on patients with substantial influence on physical and emotional health. Awareness and anticipatory counseling may be useful in stemming these effects. PMID- 11930856 TI - The emergence of women in leadership. PMID- 11930857 TI - How to stay energized in the present nursing shortage. PMID- 11930858 TI - Our penitentiaries are turning into nursing homes. PMID- 11930859 TI - The orthopedic patient's special need for wholistic caring. AB - The care of patients with musculoskeletal and subsequent orthopedic injuries is a challenging one for the nurse. Wholistic care involves much more than physical care although this is, in itself, a challenge. Care by staff, family, and friends who respond to the patient with orthopedic injuries must include attention to the complex adjustments that the patient is making psychologically to both present and future disabilities. The author's personal experience as an orthopedic patient was the impetus for this article. PMID- 11930860 TI - Who are some of the "invaluable invisible?" Some of them are licensed practical/vocational nurses. PMID- 11930861 TI - When your client has had an accident. AB - Accident victims often experience difficulty finding meaning and logic in their accident. Confusion and depression are frequently emotions that will be felt until resolution occurs. In many cases other individuals assist in this struggle for resolution. The nurse often has opportunity to provide suggestions and support as the client struggles to find meaning. PMID- 11930862 TI - A gateway to practice: the NCLEX-PN examination. PMID- 11930863 TI - Posttraumatic stress disorder: a guide for the Frontline August 2000, PTSD alliance. PMID- 11930864 TI - Maintaining intact skin during handwashing: the first line of defense against the chain of septic flow. PMID- 11930865 TI - Infectious diseases of refugees and immigrants: dengue fever. AB - PURPOSE: To describe the assessment and management of chronic wounds and detail current recommendations for optimal wound care practices. DATA SOURCES: Selected scientific literature, clinical practice guidelines, and the authors' clinical experience. CONCLUSION: Multiple factors play a role in the problems associated with chronic wounds. Wound care is costly in regards to medical care, as well as costly in terms of the hardship placed on the patient, family and staff. Too often, wound care practices are based on rituals than research. IMPLICATIONS FOR PRACTICE: The assessment and management of chronic wounds needs to be individualized according to a thorough assessment of the wound, the patients' lifestyle, and the caregiver's knowledge and skill for managing th wound. It is also important to know what wound care products are available and the types of wounds appropriate for their use. PMID- 11930866 TI - Use of PSA measurement in practice. AB - (CPG) series provides an overview of one CPG each month. The overview includes a brief summary of the guideline's content, as well as the identification of some factors by which the author has critiqued it. The first article in the series reviewed the steps of CPG critique. Subsequent columns have described CPGs related to viral upper respiratory illnesses, tobacco dependence, menopause and perimenopause, and musculoskeletal evaluation. This month, the column describes a CPG titled: Prostate-Specific Antigen (PSA) Best Practice Policy, from the American Urological Association (AUA). As prostate cancer is the leading cause of cancer deaths among U.S. men, this set of recommendations should have wide application. PMID- 11930867 TI - Congestive heart failure: understanding the pathophysiology and management. AB - PURPOSE: To explain key concepts involved in the development and management of congestive heart failure (CHF). An overview of medications commonly used in the treatment of CHF is also presented. DATA SOURCES: Selected clinical articles, major research studies, and clinical guidelines. CONCLUSIONS: The prevalence of CHF continues to increase and in the United States alone accounts for more than two million outpatient clinic visits per year at a cost of more than $10 billion annually. With the growing number of people over age 65, coupled with the advances in medical technology, primary care providers are treating many more patients with CHF. IMPLICATIONS FOR PRACTICE: A thorough understanding of the pathophysiology, especially the compensatory neurohormonal systems and ventricular remodeling that are the hallmarks of CHF, is essential for consistent application of the principles of management of this complex condition. Multidisciplinary approaches led by advanced practice nurses have excellent outcomes and improve the quality of life of patients with CHF. PMID- 11930868 TI - Wound care: meeting the challenge. PMID- 11930869 TI - Nurse practitioner patient centered telephone calls in a VA primary care geriatric clinic. AB - PURPOSE: To present original research of a retrospective study of telephone calls by nurse practitioners (NP) to geriatric primary care patients over the period of one year in a Veteran's Administration primary care clinic in the southeast. DATA SOURCES: A convenience sample of the electronic patient progress notes labeled "telephone call" of three full time NPs was collected from May 1, 1998 through April 31, 1999. A total of 1,541 unique telephone calls made by NPs to patients were examined. CONCLUSIONS: Patient telephone calls consume work time, which is often hidden and uncounted when examining time spent with patients. Most calls (62%) were generated by others; the remainder (38%) were initiated by the NPs. The greatest number (42%) of requests related to medication refills. IMPLICATIONS FOR PRACTICE: Many patients requested to talk to the NP for non-clinical issues; many of the requests could have been handled by other staff. It is as if the patient no longer has the traditional call bell to call the nurse but now has the telephone to call the NP. The personal aspect of the phone call may be what is needed by some of the patients looking for a form of human contact. PMID- 11930870 TI - Bone mineral density in postmenopausal breast cancer survivors. AB - PURPOSE: The overall purpose of this longitudinal 18-month study was to test the feasibility and effectiveness of a multicomponent intervention for prevention and treatment of osteoporosis. The purpose of this article is to describe the baseline bone mineral density (BMD) findings for 30 postmenopausal women and to compare these BMD findings to time since menopause, body mass index, and tamoxifen use. DATA SOURCES: Baseline data of BMD findings for 30 postmenopausal women, who have had a variety of treatments including surgery, adjuvant chemotherapy and or tamoxifen, and are enrolled in the 18-month longitudinal study. A demographic questionnaire and a three day dietary record were used to collect baseline data. CONCLUSIONS: Eighty percent of the women with breast cancer history had abnormal BMDs at baseline (t-scores below -1.00 SD). Thinner women showed a greater risk for accelerated trabecular bone loss at the spine and hip. IMPLICATIONS FOR PRACTICE: These findings suggest the need for early BMD assessments and for aggressive health promotion intervention strategies that include a multifaceted protocol of drug therapy for bone remodeling, 1500 mg of daily calcium, 400 IU vitamin D and a strength weight training program that is implemented immediately following chemotherapy treatment and menopause in this high risk population of women. PMID- 11930871 TI - Uncovering the facts: parental behaviors and knowledge regarding sun protection. AB - PURPOSE: A descriptive study was conducted to examine the knowledge of and behaviors related to sun-protection among parents of youth soccer players. DATA SOURCES: A convenience sample of 56 parents at community soccer events completed an 18-item instrument designed by the researchers. CONCLUSIONS: Results indicated that female respondents were more responsive to skin protection than males. In addition, advice from health care providers was shown to make an impact on the behavior of parents related to skin self-examinations and the use of sunscreen. Family history of skin cancer significantly promoted the use of protective clothing in the sun. IMPLICATIONS FOR PRACTICE: Nurse practitioners can make a difference by educating clients about sun protection and practices that can lower the risk of skin cancer and by teaching parents how to perform skin self examinations. PMID- 11930873 TI - Duplicate publication. PMID- 11930872 TI - Clinical quiz. Prepyloric heterotopic pancreas. PMID- 11930874 TI - Randomized clinical trials in hepatocellular carcinoma and biliary cancer. AB - Primary hepatocellular carcinoma (HCC) remains among the most common malignancies in the world. Many of the advances in the treatment of this disease have come from combinations of early detection in endemic areas, improved radiologic evaluation in defining extent of disease, an increased use of nonsurgical treatment and improvements in surgical technique. PMID- 11930876 TI - Safety of paracetamol packaging in the United Kingdom. PMID- 11930875 TI - Randomized clinical trials in pancreatic cancer. AB - The authors reviewed 59 prospective, randomized, controlled trials for pancreatic carcinoma that were published between 1977 and 2000. Of the 11 surgical trials, two each studied extent of resection (standard versus pylorus-preserving pancreaticoduodenectomy) and lymphadenectomy (standard versus extended lymph node dissection), five trials compared different types of pancreaticenteric reconstruction, and one each evaluated the role of prophylactic gastrojejunostomy and chemical splanchnicectomy in the setting of advanced disease. PMID- 11930877 TI - Evaluation of upper extremity motor function tests in tetraplegics. AB - OBJECTIVE: To provide an overview of arm-hand function tests useful in tetraplegic subjects. Considerations for selection of an appropriate test are also provided. DATA SOURCES: A Medline literature search was conducted covering the period from 1967 to March 2001. Relevant references cited in the selected papers were also considered, regardless of the year of publication. STUDY SELECTION: This review was restricted to strength tests, functional and ADL tests. Only general tests and tests designed specifically to test tetraplegic persons written in English, or in Dutch were included in the review. RESULTS: Information is provided on four types of strength tests, 10 general and five specific functional tests and eight ADL tests. CONCLUSION: Many tests are available to measure upper extremity motor function in tetraplegics. Selection of a test is at first determined by the outcome value in which the investigator is interested. When the type of outcome value has been determined, the most suitable test has to be selected from the range of available tests. When two tests appear to be equally suitable, the availability of information on psychometric properties of the test when used in tetraplegic patients is a decisive factor. When information on the reliability, validity and sensitivity of a test is missing, it should be gathered before using the test. PMID- 11930879 TI - The mechanism of mechanosensing by bone. PMID- 11930878 TI - Neuroendocrine immune mechanisms in rheumatic diseases. PMID- 11930881 TI - [Therapy resistant hyperlipidemia]. PMID- 11930882 TI - [Repeated facial edemas]. PMID- 11930883 TI - Comparison of entrance surface doses of some X ray examinations with CEC reference doses. AB - Entrance surface dose (ESD) measurements have been carried out in Nigeria as part of the ongoing dose reduction programme. Thermoluminescence dosemeters (TLD) were used to measure skin entrance doses for four common radiographic views in three hospitals. The mean ESD for the PA chest examination in all the participating hospitals was in the range 0.12 - 4.46 mGy. The mean ESD for the AP skull. PA skull and LAT skull were 8.55, 5.17 and 6.97 mGy respectively. The mean ESD values are greater than the CEC reference doses, except for rooms 1 and 2 in UCH where the entrance surface doses for PA chest examination are below the CEC reference dose. The QA test results show non-compliance of the accuracy of tube voltage with acceptance limit in three rooms. The timer accuracy is also not within the acceptance limit in two rooms. The reproducibility of both the kVp and timer in all the rooms is good. PMID- 11930884 TI - Assessment of infarct size by positron emission tomography and [18F]2-fluoro-2 deoxy-D-glucose: a new absolute threshold technique. AB - Along with hibernating myocardium, infarct size is a critical term in the progression of left ventricular remodelling and congestive heart failure. Both infarcted and hibernating myocardium determine changes in remote non-ischaemic tissue. This study was designed to test the accuracy of a new technique to quantify infarct size using positron emission tomography (PET) with [18F]2-fluoro 2-deoxy-D-glucose (FDG). Studies were carried out in (a) nine pigs with acute myocardial infarction (two sham-operated), produced by a 90-min occlusion of the circumflex coronary artery followed by a 4-h reperfusion, and (b) humans (six patients with ischaemic cardiomyopathy awaiting cardiac transplantation and five normal volunteers). In both animals and patients, myocardial FDG uptake was measured by PET during hyperinsulinaemic-euglycaemic clamp. Infarct size was quantified by an absolute threshold of tracer uptake obtained from the parametric (voxel-by-voxel) image of the metabolic rate of FDG. PET infarct size estimates were compared with independent ex vivo planimetric measurements of the explanted swine and patient hearts (at transplantation) after staining with triphenyltetrazolium chloride. There was good agreement between the planimetric and PET infarct size estimates both in pigs (n=9; r=0.96, v=0.94x+0.64, SEE=0.10, P<0.0001) and in humans (n=11; r=0.94, y=0.72x+2.93, SEE=0.09, P<0.0001). This study demonstrates the feasibility and accuracy of this PET method in estimating infarct size both in a model of reperfused acute myocardial infarction and in chronic ischaemic cardiomyopathy, although larger studies are needed to confirm these findings. PMID- 11930885 TI - Scintigraphic head-to-head comparison between 99mTc-WBCs and 99mTc-LeukoScan in the evaluation of inflammatory bowel disease: a pilot study. AB - Scintigraphy with technetium-99m labelled white blood cells (WBCs) is routinely used in our hospital for the assessment of inflammatory bowel disease (IBD). The main disadvantages of this diagnostic tool are its time-consuming nature and the handling of blood itself. 99mTc-LeukoScan is a relatively new, easily prepared agent that is used for the detection of osteomyelitis. To assess its value in IBD, a scintigraphic head-to-head comparison was performed between 99mTc LeukoScan and 99mTc-WBCs. 99mTc-LeukoScan scintigraphy was performed in six patients with clinically active IBD and increased uptake on 99mTc-WBC images. The interval between the scintigraphic studies ranged from 2 to 7 days, and endoscopy was subsequently performed to confirm active IBD. In three out of six patients with increased uptake on the 99mTc-WBC scans, 99mTc-LeukoScan images showed very discreet activity in the bowel, but the sites did not correspond with the inflammation sites seen on 99mTc-WBC scintigraphy and found at endoscopy. In the other three patients, 99mTc-LeukoScan scintigraphy revealed a physiological distribution but no abnormalities. In conclusion, 99mTc-LeukoScan is not an alternative agent for the assessment of IBD. A prospective study is not justified owing to the false-negative results. PMID- 11930886 TI - Unpredictability of lymphatic drainage patterns in melanoma patients. AB - We analysed the localisations of sentinel nodes (SN) found with the SN procedure to compare these sites with those that would have been predicted by conventional clinical descriptions of cutaneous lymphatic drainage. We assessed the surplus value of performing the SN procedure in melanoma patients who underwent regional nodal surgery. The SN procedure was performed in 348 patients with melanomas who were referred to our institute between 1993 and 1999. The localisations of the melanomas with the corresponding SNs were meticulously recorded on drawings of the human body and grouped according to the conventional descriptions. Predictability of lymph drainage was defined as the percentage of melanomas whose draining pattern was to the ipsilateral nearest basin, without simultaneous drainage to other basins or to an interval node. In all patients the SN procedure visualised at least one SN. We found 410 lymphatic basins in 347 patients. These basins included basins that could not have been predicted by the conventional clinical descriptions, such as multiple basins and contralateral drainage sites. For the head/neck region, SNs could be found in any of the basins described in the literature. The trunk's drainage predictability depended strongly on the melanoma localisation, ranging from 0% in the midline to 92% in one of the upper quadrants. The lower extremities had a high predictability of almost 100%, and predictability of drainage for the upper extremities ranged from 77% to 100%. In total, 34% of the patients had a cutaneous lymphatic drainage that was unpredictable, either totally or partially. We therefore conclude that an SN procedure is indispensable if the drainage site(s) are to be accurately identified. PMID- 11930887 TI - Nurses must not let patients feel unwanted. PMID- 11930889 TI - Name calling of clients is deeply ingrained. PMID- 11930890 TI - Name calling of clients is deeply ingrained. PMID- 11930888 TI - Learning disability nurses are anticipators. PMID- 11930891 TI - Coincidence timing of a tennis stroke: effects of age, skill level, gender, stimulus velocity, and attention demand. AB - Participants (N = 162, ages 10-15 years) performed a tennis stroke in a coincidence-timing task. Major results were: (a) performance improved mainly between the ages of 10-13 years; (b) skill differences were reflected by variable error (VE); (c) boys were lower than girls for VE and absolute error; (d) high stimulus velocity was related to low VE and late responding. Results concerning response organization favored explanations based on ecological perspectives rather than information processingperspectives. However, it is premature to disregard the potential role of programming. The view is supported that a continuous process of coupling perceptual and motor responses coordinates control of response timing. These cognitive processes appear to be: (a) refined with practice; (b) sensitive to differences in age, skill, and gender; and (c) adaptable to changes in environmental demands. PMID- 11930892 TI - Timing of current reproduction directly affects future reproductive output in European coots. AB - Life-history theory suggests that the variation in the seasonal timing of reproduction within populations may be explained on the basis of individual optimization. Optimal breeding times would vary between individuals as a result of trade-offs between fitness components. The existence of such trade-offs has seldom been tested empirically. We experimentally investigated the consequences of altered timing of current reproduction for future reproductive output in the European coot (Fulica atra). First clutches of different laying date were cross fostered between nests, and parents thereby experienced a delay or an advance in the hatching date. The probability and success of a second brood, adult survival until and reproduction in the next season were then compared to the natural variation among control pairs. Among control pairs the probability of a second brood declined with the progress of season. Delayed pairs were less likely and advanced pairs were more likely to produce a second brood. These changes were quantitatively as predicted from the natural seasonal decline. The number of eggs in the second clutch was positively related to egg number in the first clutch and negatively related to laying date. Compared to the natural variation, delayed females had more and advanced females had fewer eggs in their second clutch. The size of the second brood declined with season, but there was no significant effect of delay or advance. Local adult survival was higher following a delay and reduced following an advance. The effect of the experiment on adult survival was independent of sex. Laying date and clutch size of females breeding in the next year were not affected by treatment. The study demonstrates the existence of a trade-off between increased probability of a second brood and decreased parental survival for early breeding. Timing-dependent effects of current reproduction on future reproductive output may thus play an important role in the evolution of the seasonal timing of reproduction. PMID- 11930893 TI - Factors associated with success or failure in trials of vacuum extraction. AB - Failure rates for vacuum extraction of between one in 16 and one in 600 have been reported. Most studies report that unexpected failure carries a greater risk to both mother and fetus. The aim of this study was to determine factors that were likely to predict success or failure in trials of vacuum extraction. At the Port Moresby General Hospital, 59 trials of vacuum extraction were performed between 1 December 1997 and 30 November 1999. These cases were analysed according to whether vacuum extraction was achieved with more than or less than three pulls, or an alternative method of delivery was required to effect delivery Factors that were predictive of failure were: (i) Highlands origin of the mother; (ii) longer duration of the second stage of labour; (iii) severe moulding of the fetal head; (iv) cup detachments and deflexing cup applications; and (v) operator persisting with the procedure after three pulls. The cervix being less than fully dilated when the trial was commenced was not associated with a higher risk of failure, nor was it associated with a significantly higher risk of cervical trauma. Perinatal death and serious fetal scalp trauma were associated with deflexing cup applications, making more than three pulls and failed vacuum extractions. PMID- 11930894 TI - A prospective observational study on the accuracy of patient self-testing of urine at an antenatal clinic. AB - This study was performed to assess whether assigning dipstick self-testing of urine to antenatal women maintained accuracy and clinical relevance of the results. A total of 212 women were recruited from the routine antenatal clinic for the assessment of accuracy of self-testing of urine. The women's results were compared to that of the one nurse who routinely performs dipstick testing at the antenatal clinic. Analysis of the results indicates that women tended to over estimate proteinuria. PMID- 11930895 TI - Amyloid fiber formation in E. coli. PMID- 11930896 TI - Reimbursement issues surround ambulatory endoscopy centers. PMID- 11930897 TI - Improvement in xylitol production from sugarcane bagasse hydrolysate achieved by the use of a repeated-batch immobilized cell system. AB - Candida guilliermondii cells were immobilized in Ca-alginate beads and used for xylitol production from concentrated sugarcane bagasse hydrolysate during five successive fermentation batches, each lasting 48 hours. The bioconversion efficiency of 53.2%, the productivity of 0.50 g/l x h and the final xylitol concentration of 23.8 g/l obtained in the first batch increased to 61.5%, 0.59 g/l x h and 28.4 g/l, respectively, in the other four batches (mean values), with variation coefficients of up to 2.3%. PMID- 11930898 TI - Antinociceptive activity of the natural piperidine alkaloid hydrochlorides from Syphocampylus verticellatus. AB - In addition to 3'-methoxyluteolin and mixtures of sterols and triterpenes, the leaves of Syphocampylus Verticellatus yielded two piperidine alkaloid hydrochlorides, one of them has a novel structure. The alkaloids exhibit antinociceptive activity. PMID- 11930899 TI - Open clinical trial of rifabutin and clarithromycin therapy in Crohn's disease. AB - BACKGROUND: Crohn's disease, an inflammatory bowel disease in humans, has a suspected aetiology of Mycobacterium avium subsp. Paratuberculosis. AIMS: To evaluate the role of rifabutin and clarithromycin anti-Mycobacterium avium subsp. Paratuberculosis treatment in Crohn's disease patients using an open clinical trial. METHODS: . A total of 36 patients with acute presentations of Crohn's disease, whose sera tested positive against p35 and p36 antigens (two recombinant proteins of Mycobacterium avium subsp. Paratuberculosis), were selected for treatment with rifabutin and macrolide antibiotic therapy Rifabutin and macrolide antibiotic therapy medications included 250 mg 1 po bid clarithromycin and 150 mg 1 po bid Ri-fabutin accompanied with a probiotic. Crohn's disease patients' response to rifabutin and macrolide antibiotic therapy was monitored over a period ranging from 4 to 17 months. RESULTS: Seven patients (19.4%) withdrew from the study since they were unable to tolerate medications. Of the remaining 29 patients, 21 (58.3%) reached a sustained state of improvement, traditionally defined as a decrease of 70 points between their entrance and exit Crohn's disease activity index scores together with the absence of the need of all other Crohn's medications, such as immunosuppressants and corticosteroids. Three Crohn's disease patients [8. 3%) noticed significant improvements, but required other Crohn's medications, concurrently with rifabutin and macrolide antibiotic therapy, to achieve and sustain improvement. Only 5 Crohn's disease patients (13.8%) were non-responders, noticing no marked improvement while on rifabutin and macrolide antibiotic therapy. CONCLUSION: The data add further evidence to support the role of rifabutin and macrolide antibiotic therapy in the treatment of Crohn's disease specifically in those patients with evidence of Mycobacterium avium subsp. Paratuberculosis infection. A large multi-centre clinical trial is needed to further explore these findings. PMID- 11930900 TI - Familial primary biliary cirrhosis and autoimmune cholangitis. AB - AIM: Autoimmune cholangitis has been proposed as a separate disease entity from primary biliary cirrhosis without serum antimitochondrial antibodies. The ultimate answer to the question of whether autoimmune cholangitis and primary biliary cirrhosis are distinct will require detailed comparison of aetiologic factors and pathogenic mechanisms. METHODS AND RESULTS: Two families are described each of which has one member with classical antimitochondrial antibody positive biopsy-proven primary biliary cirrhosis and a first degree relative with antimitochondrial antibody negative but antinuclear antibody positive autoimmune cholangitis (biopsy proven in one case). Study of such families should allow analysis of the contribution of shared genetic risk factors versus varying environmental triggering mechanisms to disease pathogenesis. CONCLUSIONS: We suggest a European registry of families, such as the two described, which are rare within one centre, to facilitate elucidation of pathogenetic factors. PMID- 11930901 TI - Experimental hepatitis A virus (HAV) infection in Callithrix jacchus: early detection of HAV antigen and viral fate. AB - Common marmosets (Callithrixjacchus) were orally inoculated with a Brazilian strain (HAF-203) of hepatitis A virus (HAy). Three monkeys were euthanized at postinoculation hours 6, 12 and 24 to investigate the early events of HAV infection. Following others three inoculated and one control marmosets remained throughout the 46 day to evaluation of viral excretion. Different samples were collected to detect sequential presence of HAV RNA by nested reverse transcription-polymerase chain reaction (RT-PCR) in liver, saliva, bile and stools at 6 hours to 461h days postinoculation. Liver tissues were examined by immunofluorescence assay in a confocal laser-scanning microscope for the presence of HAV antigen. HAV RNA was detected in saliva during the course of the study, in bile from 24 hours to 46 days. in stools from 7 to 46 days and liver at 12 hours postinfection. In immunofluorescence of liver stained preparations, viral antigen was present at six hours after inoculation throughout the remainder of the 46-day study. The animals developed histological and biochemical acute hepatitis after second week postinoculation. Spleen, duodenum, and mesenteric lymph nodes specimens were negative for HAV antigens. This study supports the possibility that in Callithrixjacchus orally inoculated with hepatitis A virus the saliva route may be additional way of viral elimination. The viral replication in the liver was responsible for biliary HAV presence and latter HAV detection in fecal samples. PMID- 11930902 TI - Overexpression of IL-17 in the knee joint of collagen type II immunized mice promotes collagen arthritis and aggravates joint destruction. PMID- 11930904 TI - Hypersensitivity reactions in mouse airways after a single and a repeated hapten challenge. AB - OBJECTIVE AND DESIGN: In this study, we examined the effect of a single and a repeated hapten-challenge on inflammatory processes in the airways of mice undergoing a hapten-induced non-IgE mediated hypersensitivity reaction. METHODS: BALB/c mice were skin-sensitized with the hapten dinitroflourobenzene (DNFB) and intra-airway challenged with dinitrobenzene sulphonic acid (DNS). Mucosal exudation, tracheal vascular permeability, cellular accumulation, and serum murine mast cell protease (MMCP) were investigated at different time points after the first DNS-challenge and 30 min after a repeated DNS-challenge. RESULTS: MMCP levels in serum were increased at all time points after single challenge and repeated challenge. Increased vascular permeability as determined by Monastral blue staining, was found in the trachea of DNFB-sensitized mice after single DNS challenge. A second exposure to DNS profoundly enhanced the Monastral blue labeling of the tracheal blood vessels of DNFB-sensitized mice. Furthermore, increased mucosal exudation and polymorphonuclear cell (PMN) accumulation were present in DNFB-sensitized mice compared to vehicle-sensitized animals after the first DNS challenge. CONCLUSIONS: Increased mucosal exudation, vascular permeability, and PMN accumulation are prominent inflammatory features of the DNFB-induced hypersensitivity reaction in the airways. Furthermore, mast cell activation is associated with this hapten-induced hypersensitivity reaction. PMID- 11930903 TI - Gastric toxicity of racemic ketoprofen and its enantiomers in rat: oxygen radical generation and COX-expression. AB - OBJECTIVE AND DESIGN: The gastric toxicity of racemic-ketoprofen and its enantiomers (S(+)- and R(-)-ketoprofen), oxygen free radical generation and neutrophil infiltration in response to damage were evaluated in rats. Changes in prostaglandin synthesis, cyclooxygenase expression and glutathione metabolism were also studied. MATERIALS AND METHODS: Studies were performed in Wistar rats. Drugs were given by oral administration: racemic-ketoprofen (100, 50 and 25 mg/kg body weight); S(+) and R(-)-ketoprofen (50, 25 and 12.5 mg/kg body weight). Determinations were made of gastric mucosal injury, lipid peroxidation, xanthine oxidase, myeloperoxidase and superoxide dismutase activities, glutathione levels, glutathione peroxidase and glutathione reductase activities, gastric prostaglandin synthesis (PGE2 levels) and COX-expression. RESULTS: Racemic ketoprofen dose-dependently exhibited the highest toxicity. In contrast, S(+) ketoprofen at half the dose of the racemic compound proved to be less ulcerogenic. R(-)-ketoprofen was also less ulcerogenic, but when administered as the racemate exacerbated gastric ulceration caused by S(+)-ketoprofen. Drug administration produced significant increases in lipid peroxidation levels and xanthine-oxidase and a decrease in superoxide dismutase activity. Nevertheless the racemate induced the highest disturbances in oxidative metabolism. No changes in myeloperoxidase values and glutathione metabolism were found. Cyclooxygenase-1 immunoreactivity was observed and did not differ from that in control rats. Cyclooxygenase-2 could also be expressed after treatments. CONCLUSIONS: R(-) ketoprofen and S(+)-ketoprofen have a comparable gastric toxicity and they both have a better gastric toxicity profile as compared to the racemate. In addition to inhibition of prostaglandin synthesis, damage resulted in an increase of cyclooxygenase-2 protein expression. Oxygen radicals, including superoxide anions, could also be implicated. PMID- 11930905 TI - Iterative design and evaluation of new prone carts for individuals with SCDs: a technical note. AB - This paper summarizes a series of projects funded since 1992 to address the compelling need to improve the quality of life for persons with spinal cord dysfunctions who use prone carts. Specifically, Veterans Services Organization, National Institute on Disability and Rehabilitation Research (NIDRR), and the VA Rehabilitation Research and Development funded studies to develop new consumer driven designs for prone carts. Using an iterative approach, this team of clinicians and designers (1) evaluated existing prone carts; (2) designed a new manual prone cart; (3) designed a new motorized prone cart, including a standing model; and (4) are collaborating with manufacturers to market and commercialize the new prone carts. Prototypes were developed at the Milwaukee Institute of Art and Design with the assistance of Ortho-Kinetics, Inc., and Everest & Jennings and were clinically evaluated at two Veterans Affairs Medical Centers (Tampa and Milwaukee) with patients and caregivers and for compliance with applicable ISO (International Organization for Standardization) for electric wheelchair standards. PMID- 11930906 TI - Normative values and determinants of physical capacity in individuals with spinal cord injury. AB - We reanalyzed data from five studies with similar or identical methodology performed by our laboratory to define normative values and determinants of physical capacity in individuals with tetraplegia and paraplegia. Each study consisted of a graded wheelchair exercise test to determine peak oxygen uptake and maximal power output and could additionally include a wheelchair sprint test to determine short-term (anaerobic) power output and/or an isometric strength test. The combined subject population included 166 individuals (20 women), varying considerably for age, body mass, lesion level, time since injury, and activity level. Ranges in physical capacity parameters were extensive and normative values for individuals with tetraplegia and paraplegia were established. These physical capacity norms could be used for evaluation of fitness status and training or therapeutic interventions. Multiple regression procedures indicated that 48-80% of the variance in physical capacity could be explained by lesion level and completeness, activity level, gender, age, body mass, and time since injury. Although physical capacity is largely determined by factors that cannot be altered, such as lesion level, age, and gender, changeable factors such as activity level and body mass play an additional role. PMID- 11930907 TI - Oral mucosa: variations from normalcy, part II. AB - This is the second article in a 2-part series on the variations of oral mucosa. We describe the following 5 conditions that deviate from normalcy: lateral soft palate fistulas, double lip, fissured tongue, racial gingival pigmentation, and geographic tongue. PMID- 11930908 TI - Up-regulation of 5-HT2B receptor density and receptor-mediated glycogenolysis in mouse astrocytes by long-term fluoxetine administration. AB - The effects were studied of short-term (1 week) versus long-term (2-3 weeks) fluoxetine treatment of primary cultures of mouse astrocytes, differentiated by treatment with dibutyryl cyclic AMP. From previous experiments it is known that acute treatment with fluoxetine stimulates glycogenolysis and increases free cytosolic Ca2+ concentration ([Ca2+]i]) in these cultures, whereas short-term (one week) treatment with 10 microM down-regulates the effects on glycogen and [Ca2+]i, when fluoxetine administration is renewed (or when serotonin is administered). Moreover, antagonist studies have shown that these responses are evoked by activation of a 5-HT2, receptor that is different from the 5-HT2A receptor and therefore at that time tentatively were interpreted as being exerted on 5-HT2C receptors. In the present study the cultures were found by RT-PCR to express mRNA for 5-HT2A and 5-HT2B receptors, but not for the 5-HT2C receptor, identifying the 5-HT2 receptor activated by fluoxetine as the 5-HT2B receptor, the most recently cloned 5-Ht2 receptor and a 5-HT receptor known to be more abundant in human, than in rodent, brain. Both short-term and long-term treatment with fluoxetine increased the specific binding of [3H]mesulergine, a ligand for alL three 5-HT2 receptors. Long-term treatment with fluoxetine caused an agonist induced up-regulation of the glycogenolytic response to renewed administration of fluoxetine, whereas short-term treatment abolished the fluoxetine-induced hydrolysis of glycogen. Thus, during a treatment period similar to that required for fluoxetine's clinical response to occur, 5-HT2B-mediated effects are initially down-regulated and subsequently up-regulated. PMID- 11930909 TI - Alzheimer's disease research enters a "new cycle": how significant? AB - Recent evidence suggests a link between the aberrant re-expression of cell cycle proteins in adult neurons of the Alzheimer's disease brain and the process of apoptotic degeneration. Here we will discuss this unexpected phenomenon as related to the mechanisms of beta-amyloid toxicity, and its significance for therapeutic possibilities. PMID- 11930910 TI - A comparison of some metabolic effects of N-methylaspartate stereoisomers, glutamate and depolarization: a multinuclear MRS study. AB - Exposure of guinea pig brain slices to low concentrations (10 microM) of NMDA caused decreases in PCr and ATP within 30 min, with a slower decrease in NAA and increase in lactate, both detectable after 1 h. Exposure to NMDA for over 1 h or at higher concentrations caused further increases in lactate and decreases in NAA, with no further change in PCr or ATP. The L-isomer, NMLA, and the racemic mixture, NMDLA, caused similar changes in lactate and NAA, but both produced greater decreases in the energy state than NMDA, similar to those caused by prolonged exposure to glutamate. MK-801 prevented the changes in the energy state caused by NMDA, but not those caused by NMLA or by glutamate. The results are compared to previous studies on depolarization and discussed in terms of the role of the NMDA sub-type of glutamate receptor in the excitotoxic hypothesis of neuronal degeneration. PMID- 11930911 TI - Role of ET(A) receptors in the vasoconstriction induced by endothelin-1 in subcutaneous small arteries of normotensive subjects and hypertensive patients. AB - OBJECTIVE: The aim of our study was to investigate contractile responses to endothelin-1 in the presence or absence of selective blockers of ET(A) or ET(B) receptors in subcutaneous small resistance arteries of normotensive subjects and of patients with essential hypertension. METHODS: Twenty-four subjects (eight normotensives aged 50 +/- 4 years, and 16 with essential hypertension aged 53 +/- 4 years) were included in the study. All subjects were submitted to a biopsy of the subcutaneous fat. Small resistance arteries (internal diameter 160-280 microm) were dissected and mounted on a micromyograph as a ring preparation (Mulvany's technique). The media-to-lumen ratio was calculated. A concentration response curve to endothelin-1 was then performed in the presence or absence of FR 139317, (a selective blocker of ET(A) receptors) or of BQ 788, (a selective blocker of ET(B) receptors). RESULTS: The media-to-lumen ratio was lower in normotensives compared with those subjects with essential hypertension (0.08 +/- 0.02 vs. 0.12 +/- 0.05, p < 0.01). The vasoconstriction induced by endothelin-1 was greater in normotensives than in patients with essential hypertension. In normotensives, almost all the vasoconstriction induced by endothelin-1 was blocked by the addition of FR 139317, while in subjects with essential hypertension the effect was smaller. The selective blocker of ET(B) was devoid of effect in both groups. CONCLUSIONS: The vasoconstrictor effect of endothelin-1 in small resistance arteries of normotensive subjects and, in part, also in hypertensive patients is mediated by ET(A) receptors, while ET(B) receptors play a minor role, if any. It is, however, possible that a vasoconstrictor effect mediated by ET(B) receptors located on vascular smooth muscle cells may be masked by the simultaneous stimulation of endothelial ET(B) receptors which may induce a vasodilation mediated by nitric oxide. PMID- 11930912 TI - Quality assurance in IMRT: importance of the transmission through the jaws for an accurate calculation of absolute doses and relative distributions. AB - The goal of IMRT is to achieve an isodose distribution conformed to the tumor while avoiding the organs at risk. For these tasks several gantry angles are selected, each one containing a series of different leaf configurations for the multileaf collimator (MLC) (segments). Verifying the relative distributions as well as the absolute doses is an important step for quality assurance issues. We have observed that an accurate modeling of the transmission of the primary x-ray fluence through the jaws and MLC as well as the head scatter is crucial for a precise calculation of relative doses and monitor units. Also, an inaccurate calculation of the output factor for small size segments can lead to important differences in the absolute dose for points under these segments. Incorrect models could lead to systematic errors of around 5% to 10% in the calculated monitor units and a shift in the isodose curves. PMID- 11930913 TI - Sensitivity of megavoltage photon beam Monte Carlo simulations to electron beam and other parameters. AB - The BEAM code is used to simulate nine photon beams from three major manufacturers of medical linear accelerators (Varian, Elekta, and Siemens), to derive and evaluate estimates for the parameters of the electron beam incident on the target, and to study the effects of some mechanical parameters like target width, primary collimator opening, flattening filter material and density. The mean energy and the FWHM of the incident electron beam intensity distributions (assumed Gaussian and cylindrically symmetric) are derived by matching calculated percentage depth-dose curves past the depth of maximum dose (within 1% of maximum dose) and off-axis factors (within 2sigma at 1% statistics or less) with measured data from the AAPM RTC TG-46 compilation. The off-axis factors are found to be very sensitive to the mean energy of the electron beam, the FWHM of its intensity distribution, its angle of incidence, the dimensions of the upper opening of the primary collimator, the material of the flattening filter and its density. The off-axis factors are relatively insensitive to the FWHM of the electron beam energy distribution, its divergence and the lateral dimensions of the target. The depth-dose curves are sensitive to the electron beam energy, and to its energy distribution, but they show no sensitivity to the FWHM of the electron beam intensity distribution. The electron beam incident energy can be estimated within 0.2 MeV when matching either the measured off-axis factors or the central-axis depth-dose curves when the calculated uncertainties are about 0.7% at the 1 sigma level. The derived FWHM (+/-0.1 mm) of the electron beam intensity distributions all fall within 1 mm of the manufacturer specifications except in one case where the difference is 1.2 mm. PMID- 11930914 TI - Monte Carlo calculation of nine megavoltage photon beam spectra using the BEAM code. AB - A recent paper analyzed the sensitivity to various simulation parameters of the Monte Carlo simulations of nine beams from three major manufacturers of commercial medical linear accelerators, ranging in energy from 4-25 MV. In this work the nine models are used: to calculate photon energy spectra and average energy distributions and compare them to those published by Mohan et al. [Med. Phys. 12, 592-597 (1985)]; to separate the spectra into primary and scatter components from the primary collimator, the flattening filter and the adjustable collimators; and to calculate the contaminant-electron fluence spectra and the electron contribution to the depth-dose curves. Notwithstanding the better precision of the calculated spectra, they are similar to those calculated by Mohan et al. The three photon spectra at 6 MV from the machines of three different manufacturers show differences in their shapes as well as in the efficiency of bremsstrahlung production in the corresponding target and filter combinations. The contribution of direct photons to the photon energy fluence in a 10 x 10 field varies between 92% and 97%, where the primary collimator contributes between 0.6% and 3.4% and the flattening filter contributes between 0.6% and 4.5% to the head-scatter energy fluence. The fluence of the contaminant electrons at 100 cm varies between 5 x 10(-9) and 2.4 x 10(-7) cm(-2) per incident electron on target, and the corresponding spectrum for each beam is relatively invariant inside a 10 x 10 cm2 field. On the surface the dose from electron contamination varies between 5.7% and 11% of maximum dose and, at the depth of maximum dose, between 0.16% and 2.5% of maximum dose. The photon component of the percentage depth-dose at 10 cm depth is compared with the general formula provided by AAPM's task group 51 and confirms the claimed accuracy of 2%. PMID- 11930915 TI - Serum carotenoids and clustering of adverse health behaviors in American adults. AB - OBJECTIVE: To determine the association between clustering of adverse health behaviors (cigarette smoking, alcohol intake, and sedentarism) and serum carotenoids in a representative sample of white, Black, and Hispanic Americans. METHODS: Data (n=6,218) from the Third US National Health and Nutrition Examination Survey were utilized for this investigation. Adjusted gender-specific linear regression analyses were used. RESULTS: Increase in clustering of adverse health behaviors was associated with decreased serum concentrations of a carotene, b-carotene, bcryptoxanthine, and luteine/zeaxanthene in the 3 racial/ethnic groups (P<0.05). CONCLUSION: Persons with multiple adverse health behaviors should be advised to increase fruit and vegetable consumption, as primary sources of chemopreventive antioxidants. PMID- 11930916 TI - Antibiotic resistance from down on the chicken farm. PMID- 11930918 TI - A new kind of fish story. The coming of biotech animals. PMID- 11930917 TI - Fighting the flu. PMID- 11930919 TI - Heading off hair-care disasters. Use caution with relaxers and dyes. PMID- 11930920 TI - The international flow of food. FDA takes on growing responsibilities for imported food safety. PMID- 11930921 TI - Glaxo Wellcome withdraws irritable bowel syndrome medication. PMID- 11930922 TI - Dietary supplement maker fined twice what company profited. PMID- 11930923 TI - Hearing the cry for help and information. PMID- 11930924 TI - Drug helps delay progression of multiple sclerosis. PMID- 11930925 TI - Arsenic-based therapy benefits leukemia patients. PMID- 11930926 TI - Combination drug approved to treat HIV. PMID- 11930927 TI - New uses for old drug: reducing risk of heart attacks and strokes. PMID- 11930928 TI - Monthly injection provides new contraceptive choice. PMID- 11930929 TI - Survey shows parents need to measure children's medicine more accurately. PMID- 11930930 TI - New glue approved to reduce blood flow during brain surgery. PMID- 11930931 TI - Electronic brain implant helps patients regain hearing after cranial nerve surgery. PMID- 11930932 TI - Device delivers shock waves to help ease heel pain caused by plantar fasciitis. PMID- 11930933 TI - Head injury linked to increased risk of Alzheimer's disease. PMID- 11930934 TI - FDA issues public health advisory on phenylpropanolamine in drug products. PMID- 11930935 TI - Quokkas (Setonix brachyurus) demonstrate tactile discrimination learning and serial-reversal learning. AB - Two male quokkas (Setonix brachyurus: a herbivorous macropod marsupial) were trained to discriminate pairs of stimuli in the laboratory. Quokkas indicated their choice by pulling on 1 of 2 simultaneously presented cords. The quokkas' discrimination abilities were tested on 6 tactile and 6 visual discrimination tasks. Correct responses were rewarded with food. For both quokkas, all tactile tasks were learned to a criterion of 75% correct in up to 4 20-trial sessions. No visual task maintained criterion performance in 4 sessions. One tactile discrimination was reversed 10 times. After the 1st reversal, the error rate declined sharply and fell to a level well below the initial discrimination. PMID- 11930936 TI - Visual discrimination learning and strategy behavior in the fat-tailed dunnart (Sminthopsis crassicaudata). AB - Fat-tailed dunnarts (Sminthopsis crassicaudata) were trained on visual discrimination learning-set, reversal-set, and spatial delayed-alternation tasks. The learning set involved 36 2-way black-and-white pattern discriminations and 5 probe reversals. Ten reversals of a black-and-white pattern discrimination were followed by 5 novel tasks. Spatial alternation was tested at delays up to 20 s. Learning-set and reversal-set formation, including 1-trial learning and spontaneous transfer from learning set to reversals and vice versa, was found. Learning-set-experienced dunnarts showed no retention of previously learned tasks 1 week after testing but demonstrated consistently high Trial 2 performance, indicating the retention of a response strategy. Delayed-alternation tasks were learned up to 10-s delays. These results provide the first evidence of a visually guided "win-stay, lose-shift" strategy in a marsupial. PMID- 11930937 TI - An index of relationship quality based on attachment theory. AB - Two measures are reported of the nature or quality of a mother-offspring (MO) relationship during development using brown capuchin monkeys (Cebus apella) as models. One is a qualitative classification of MO relationships as secure, resistant, or avoidant attachments. The other is an empirical ratio of relative affiliation to agonism called the MO relationship quality, or MORQ, Index. The two methods tapped similar relationship features so relationships high or low of a median split of MORQ values were heuristically labeled secure (n = 22) or insecure (n = 16), respectively. A comparison revealed extensive behavioral differences between secure and insecure MO relationships and suggested MORQ provided an objective, continuous measure of attachment security. PMID- 11930938 TI - Activation of p38 MAP-kinase and caldesmon phosphorylation are essential for urokinase-induced human smooth muscle cell migration. AB - We have explored intracellular pathways involved in the urokinase type plasminogen activator (urokinase or uPA)-stimulated migration of human airway smooth muscle cells (hAWSMC). Using a set of uPA mutants we found that protease activity, growth factor-like and kringle domains of uPA differentially contribute to activation of p42/p44erk1,2 and p38 MAP-kinases. Consistent with our earlier data [Mukhina et al., J. Biol. Chem. 275 (2000), 16450-16458], the kringle domain of uPA was sufficient and required to stimulate cell motility. Here we report that uPA mutants containing the kringle domain specifically activate the p38 MAP kinase pathway and actomyosin by increasing phosphorylation of the critical Ser 19 on the myosin regulatory light chain and MAP-kinase sites of the actin associated regulatory protein caldesmon. While pharmacological inhibition of p38 MAP-kinase activation did not affect myosin light chain phosphorylation, it blocked the increase in caldesmon phosphorylation and uPA-stimulated migration of hAWSMC on a collagen-coated surface. We conclude that activation of p38 MAP kinase and downstream phosphorylation of non-muscle caldesmon is essential for urokinase-stimulated smooth muscle cell migration. PMID- 11930940 TI - The effect of respiratory tract infections on reported asthma symptoms. AB - BACKGROUND: Wheezing and chest tightness is associated with respiratory tract infections. Thus the occurrence of respiratory tract infections may influence the prevalence of asthma. AIMS: To assess the strength of relation between two main symptoms of asthma and the prevalence of respiratory tract infections in children with and without asthma. METHODS: The study population was 3,796 children, four years of age, whose parents had answered a questionnaire on respiratory symptoms and asthma. The prevalence of wheezing and chest tightness in the last 12 months and four weeks was estimated according to the occurrence of respiratory tract infections during the corresponding time periods; 95% confidence intervals of the prevalence and the statistical significance of the differences in the prevalence by using chi-square test were calculated. RESULTS: The prevalence of wheezing and chest tightness was higher in children who had experienced respiratory tract infections than in those who had not. The prevalence of wheezing was 17.5% among children who had experienced bronchitis and 3.0% among children whom had not (p<0.001). The prevalence of chest tightness in the last 12 months was 3.7% (95% CI 3.1-4.1) among children without asthma and 59.3% (95% CI 53.4-65.2) among children with asthma (p<0.001). The prevalence of wheezing and chest tightness increased with increasing number of different types of respiratory tract infections among children without asthma (p<0.001) and tended to be so among children with asthma. CONCLUSIONS: These findings have implications for the interpretation of results from epidemiological studies using respiratory symptoms as a health-related outcome of asthma. PMID- 11930939 TI - uPA-silica-Particles (SP-uPA): a novel analytical system to investigate uPA-uPAR interaction and to test synthetic uPAR antagonists as potential cancer therapeutics. AB - The urokinase-type plasminogen activation system, including the serine protease uPA (urokinase-type plasminogen activator) and its cell surface receptor (uPAR, CD87), are important key molecules in tumor invasion and metastasis. Besides its proteolytic function, binding of uPA to uPAR on tumor cells exerts various cell responses such as migration, adhesion, proliferation, and differentiation. Hence, the uPA/uPAR system is a potential target for tumor therapy. We have designed a new generation of uPA-derived synthetic cyclic peptides suited to interfere with the binding of uPA to uPAR and present a new technology involving micro silica particles coated with uPA (SP-uPA) and reacting with recombinant soluble uPAR (suPAR), to rapidly assess the antagonistic potential of uPA-peptides by flow cytofluorometry (FACS). For this, we used silica particles of 10 microm in diameter to which HMW-uPA is coupled using the EDC/NHS method. Soluble, recombinant suPAR was added and the interaction of SP-uPA with suPAR verified by reaction with monoclonal antibody HD13.1 directed to uPAR, followed by a cyan dye (cy5)-labeled antibody directed against mouse IgG. Thereby it was possible to test naturally occurring ligands of uPAR (HMW-uPA, ATF) as well as highly effective, synthetic cyclic uPA-derived peptides (cyclo21,29[D-Cys21Cys29]-UPA21 30, cyclo21,29[D-Cys21Nle28Cys29]-uPA21-30, cyclo21,29[D-Cys(21)2-Nal24Cys29] uPA21-30, and cyclo21,29[D-Cys21Orn23Thi24Thi25Cys29]-uPA21-30. The results obtained with the noncellular SP-uPA/uPAR system are highly comparable to those obtained with a cellular system involving FITC-uPA and the promyeloid cell line U937 as the source of uPAR. PMID- 11930941 TI - Local fibrinolytic therapy with urokinase for peritoneal dialysis catheter obstruction in children. PMID- 11930942 TI - Consensus on peritonitis treatment in pediatric patients? PMID- 11930944 TI - Environmental air degradation in Dar es Salaam by x-ray fluorescence. AB - In Dar es Salaam a study of the aerosol contents was conducted and particulate matter on the filters were collected using an Andersen PM10 impactor to determine the environmental air pollution. The contents were determined by X-ray fluorescence analysis. In this study sources of environmental degradation and the concentrations were named as follows: Combustion processes with range of Br from 10 to 800 ng/m3, Pb from 30 to 790 ng/m3. Industrial processes with range of Fe from 37 to 883 ng/m3, Cu from 14 to 310 ng/m3, Zn from 6 to 820ng/m3. Top soil activities with range of K from 20 to 540 ng/m3, Ca from 24 to 3805 ng/m3, Ti from 2 to 59 ng/m3, Mn from 10 to 386 ng/m3. Marine processes with range of Cl from 20 to 310 ng/m3, S from 72 to 134 ng/m3. PMID- 11930943 TI - Extracellular beta-D-glucosidase from Chaetomium thermophilum var. coprophilum: production, purification and some biochemical properties. AB - The thermophilic fungus Chaetomium thermophilum var. coprophilum produced large amounts of extracellular and intracellular beta-glucosidase activity when grown on cellulose or cellobiose as carbon sources. The presence of glucose in the culture medium drastically decreased the level of beta-glucosidase activity, while cycloheximide prevented the induction of the extracellular enzyme activity by cellobiose. An extracellular beta-glucosidase induced by avicel was purified by a procedure involving acetone precipitation and chromatography on two DEAE cellulose columns. The purified enzyme was a basic protein, with a carbohydrate content of 73%. The deglycosylated enzyme exhibited a molecular mass of 43 kDa, with pH and temperature optima of 5.5 and 65 degrees C respectively. The beta glucosidase hydrolysed only cellobiose and p-nitrophenyl-beta-D-glucopyranoside, exhibiting apparent Km values of 3.13 mM and 0.76 mM, respectively. The native purified enzyme was stable up to 2 hours at 60 degrees C, and its thermal stability was directly dependent on glycosylation. PMID- 11930945 TI - Is 5-aminolevulinic acid involved in the hepatocellular carcinogenesis of acute intermittent porphyria? AB - 5-Aminolevulinic acid (ALA) is a heme precursor that accumulates in acute intermittent porphyria (AIP) due to enzymatic deficiencies in the heme biosynthetic pathway Its accumulation has been associated with several symptoms, such as abdominal pain attacks, neuromuscular weaknesses, neuropsychiatric alterations and increased hepatocellular carcinoma (HCC) incidence. The use of exogenous ALA to elevate porphyrin levels in tumor photodynamic therapy, adds further significance to ALA toxicology. Under ferritin mediated and metal catalyzed oxidation, ALA produces reactive oxygen species that can damage plasmid and isolated DNA in vitro, and increases the steady-state level of 8-oxo-7,8 dihydro-2'-deoxyguanosine in liver, spleen and kidney DNA and 5-hydroxy-2' deoxycytidine in liver DNA of ALA-treated rats. The in vitro DNA damage could be partially inhibited by SOD, catalase, DTPA, mannitol and melatonin. ALA also promotes the formation of radical-induced base degradation products in isolated DNA. 4,5-Dioxovaleric acid, the final oxidation product of ALA, alkylates guanine moieties within both nucleoside and isolated DNA, producing two diastereoisomeric adducts. Dihydropyrazine derivatives of ALA generated by its dimerization, promote DNA strand-breaks and 8-oxodGuo formation in the presence of Cu2+. Together these results reinforce the hypothesis that the DNA damage induced by ALA may be associated with the development of HCC in individuals suffering from AIP. PMID- 11930946 TI - Haplotype analysis excludes the functional protoporphyrinogen oxidase promoter polymorphism -1081G>A as a modifying factor in the clinical expression of variegate porphyria. AB - Variegate porphyria (VP) is caused by the founder-type protoporphyrinogen oxidase (PPOX) gene mutation R59W in the majority of South African patients. VP is inherited as an autosomal dominant disease with incomplete penetrance and no genotype-phenotype association has been established to date. In an attempt to determine whether a relatively common mutation in the promoter region of the gene (-1081G>A) represents a low-expression allele that may influence clinical manifestation of the disease when inherited from the non-carrier (R59W-negative) parent, we have studied the effect of the mutated allele using an in vitro luciferase assay. Haplotype analysis was furthermore used to evaluate the added information obtained by considering the possible influence of this mutation in combination with a polymorphism in intron 2 (206G>C) of the gene in a genotype phenotype correlation study. Although the mutation at nucleotide -1081 resulted in a significant reduction in transcriptional activity relative to the reference wild type, no evidence could be obtained that a specific haplotype inherited from the normal parent affects clinical expression of the disease. We thus conclude that other factors such as modifier loci unrelated to the PPOX gene may determine clinical manifestation of VP. PMID- 11930947 TI - Is a 0900-h serum cortisol useful prior to a short synacthen test in outpatient assessment? AB - BACKGROUND: The short synacthen test (SST) is the gold standard investigation for the evaluation of adrenal insufficiency and is also frequently used for the evaluation of the hypothalamic-pituitary-adrenal (HPA) axis. The 0900-h serum cortisol concentration has also been evaluated as an indication of cortisol reserve, and a result > 450 nmol/L is highly suggestive of a normal serum cortisol response to the insulin tolerance test, while no patient with a 0900-h serum cortisol < 100 nmol/L had a sufficient response. The aim of this study was to determine if the number of inappropriate SSTs could be reduced if a 0900-h serum total cortisol was done prior to the dynamic function test. METHOD: Two hundred and ten SSTs were performed at 0900 h and the response at 30 min evaluated. RESULTS: Of the 210 SST, 151 (71%) demonstrated a maximum response at 30 min of serum cortisol > 550 nmol/L. All the patients with a 0900-h serum cortisol > 500 nmol/L had an adequate response ( > 550 nmol/L), while no patient with a 0900-h serum cortisol of < 100 nmol/L had an adequate SST response. Twenty one per cent of patients were shown to have had an unnecessary invasive procedure. CONCLUSION: We conclude that the SST is of little added value in patients with a 0900-h serum cortisol of less than 100 nmol/L or more than 500 nmol/L and it should be included in the appropriate protocols for endocrine investigation. PMID- 11930948 TI - Impaired fibrinolysis in premenopausal women and age-matched men with Type 2 diabetes mellitus: a pilot study. AB - BACKGROUND: The presence of Type 2 diabetes mellitus (DM) is one of the strongest predictors of cardiovascular disease (CVD) in women. Although the specific mechanisms underlying this increased risk are unknown, one factor that may contribute to CVD in women with Type 2 DM is impaired fibrinolysis. Healthy premenopausal women have a low rate of CVD and excellent fibrinolytic potential. Impairment in fibrinolysis in people with DM has been demonstrated mainly in men, whereas the fibrinolytic potential of women with Type 2 DM has not been characterized well. This pilot study compared fibrinolytic measures in premenopausal women and men with DM with those of healthy age-matched control women and men to help determine whether fibrinolysis is abnormal in women with DM. METHODS: Fibrinolytic measurements included euglobulin clot lysis time (ELT), fibrinogen, plasminogen activator inhibitor 1, and tissue-type plasminogen activator. RESULTS: Poststasis ELT was significantly impaired in the women with DM as compared with the control women. The men with DM had a tendency toward slower poststasis ELT than did the control men, but the differences between the men's groups were not significant. In the women's groups only, we observed a trend toward increased plasminogen activator inhibitor 1 among the women with DM. CONCLUSIONS: Women with DM have a more significant abnormality in poststasis ELT than do men with DM as compared with sex-specific counterparts without DM. PMID- 11930949 TI - Safety evaluation of routine intracoronary acetylcholine infusion in patients undergoing a first diagnostic coronary angiogram. AB - BACKGROUND: Recent findings imply prognostic significance of intracoronary acetylcholine infusion for endothelial function testing. We evaluated whether routine use of this test in coronary angiography patients is safe. METHODS: Patients undergoing a first diagnostic coronary angiography were selected to receive intracoronary acetylcholine for endothelial function evaluation. The relation between adverse reactions during infusion and risk factors was analyzed with a logistic regression model. Included in the multiple logistic regression model were the variables with a univariate P value < 0.20. RESULTS: Adverse reactions occurred in 16% (49/299) of the patients. This included two life threatening events caused by occlusive spasm and flow limitation in the left coronary artery. Other adverse events were chest pain (n = 38), AV block or sinus bradycardia (n = 10), dyspnea (n = 3). Adverse reactions were more likely to occur in patients younger than 60 years of age (relative risk, 5.6 [2.2-14.3]). CONCLUSION: Intracoronary acetylcholine infusion is safe, but may lead to serious adverse reactions. Care should be taken especially in patients younger than 60 years of age. Routine use of acetylcholine infusion can thus only be justified if it has important prognostic significance. This has to be proven further in large prospective studies. PMID- 11930950 TI - Prediction of the adherence, growth and release of microorganisms in production chains. AB - The aim of this study was to develop a mathematical model that describes the bacterial contamination of food as a result of adherence, growth and release of bacteria in process equipment. The model developed can be applied to control the bacterial quality of food products produced in process chains in which the final contamination of the product is governed by growth and heat-induced destruction of bacteria. To set up the model, experiments were carried out with a plate heat exchanger using milk inoculated with Streptococcus thermophilus. The growth rate of S. thermophilus in milk could be described accurately by the modified expanded model of Ratkowsky. The observed increase in the concentration of S. thermophilus in milk at the outlet of the plate heat exchanger could be described quantitatively by the model. To predict the contamination of the product, the model was integrated into NIZO-PCS (Process Chain Simulator). The results of computer simulations were validated by a number of measurements in a cheese factory. It turned out that the agreement between the measured and calculated concentrations of S. thermophilus was sufficient for the model to be used for predictions in industrial production chains. PMID- 11930952 TI - Inactivation of conidia of Botrytis cinerea and Monilinia fructigena using UV-C and heat treatment. AB - The effect of UV-C (lambda = 254 nm) and heat treatment was investigated on the inactivation of conidia of Botrytis cinerea and Monilinia fructigena, two major postharvest spoilage fungi of strawberries and cherries, respectively. Both fungi were grown at 21 degrees C in the dark and conidia were isolated after 1 week by washing the mycelium with a mild detergent solution. After filtration and resuspension in phosphate buffer to a titer of 10(5) to 10(6) cfu/ml, the conidia were subjected to different treatments. The applied UV-C doses varied from 0.01 to 1.50 J/cm2, and the conditions for the thermal treatment were 3, 5, 10, 15 and 20 min at temperatures ranging from 35 to 48 degrees C. Both techniques were applied individually and in combination. Spore inactivation increased with increasing intensity of single treatments. No surviving spores of B. cinerea were observed after 15 min at 45 degrees C or an UV-C treatment of 1.00 J/cm2. M. fructigena was more sensitive and a thermal treatment of 3 min at 45 degrees C or an UV-C treatment of 0.50 J/cm2 resulted in complete spore inactivation. Combination of both techniques reduced the required intensity of the treatment for inactivation of both fungi. The order of the applications had a significant effect on the degree of inactivation. The inactivation of B. cinerea conidia was greater when the heat treatment came first, and for M. fructigena, most inactivation was achieved when the heat treatment was preceded with an UV-C irradiation. PMID- 11930951 TI - On the use of the Weibull model to describe thermal inactivation of microbial vegetative cells. AB - This paper evaluates the applicability of the Weibull model to describe thermal inactivation of microbial vegetative cells as an alternative for the classical Bigelow model of first-order kinetics; spores are excluded in this article because of the complications arising due to the activation of dormant spores. The Weibull model takes biological variation, with respect to thermal inactivation, into account and is basically a statistical model of distribution of inactivation times. The model used has two parameters, the scale parameter alpha (time) and the dimensionless shape parameter beta. The model conveniently accounts for the frequently observed nonlinearity of semilogarithmic survivor curves, and the classical first-order approach is a special case of the Weibull model. The shape parameter accounts for upward concavity of a survival curve (beta < 1), a linear survival curve (beta = 1), and downward concavity (beta > 1). Although the Weibull model is of an empirical nature, a link can be made with physiological effects. Beta < 1 indicates that the remaining cells have the ability to adapt to the applied stress, whereas beta > 1 indicates that the remaining cells become increasingly damaged. Fifty-five case studies taken from the literature were analyzed to study the temperature dependence of the two parameters. The logarithm of the scale parameter alpha depended linearly on temperature, analogous to the classical D value. However, the temperature dependence of the shape parameter beta was not so clear. In only seven cases, the shape parameter seemed to depend on temperature, in a linear way. In all other cases, no statistically significant (linear) relation with temperature could be found. In 39 cases, the shape parameter beta was larger than 1, and in 14 cases, smaller than 1. Only in two cases was the shape parameter beta = 1 over the temperature range studied, indicating that the classical first-order kinetics approach is the exception rather than the rule. The conclusion is that the Weibull model can be used to model nonlinear survival curves, and may be helpful to pinpoint relevant physiological effects caused by heating. Most importantly, process calculations show that large discrepancies can be found between the classical first-order approach and the Weibull model. This case study suggests that the Weibull model performs much better than the classical inactivation model and can be of much value in modelling thermal inactivation more realistically, and therefore, in improving food safety and quality. PMID- 11930953 TI - The enumeration and identification of acetic acid bacteria from South African red wine fermentations. AB - Acetic acid bacteria are microorganisms that can profoundly influence the quality of wine. Surprisingly, little research has been done on these microorganisms in the winemaking field. The object of this study was to investigate the occurrence of acetic acid bacteria in South African red wine fermentations and to identify the dominant species occurring. Acetic acid bacteria were isolated and enumerated from small-scale and commercial red must fermentations in 1998 and 1999, respectively. The initial occurrence of acetic acid bacteria in the must was shown to vary with cell numbers ranging from 10(6)-10(7) to 10(4)-10(5) cfu/ml for the 1998 and 1999 musts, respectively. The acetic acid bacteria decreased to 10(2)-10(3) cfu/ml in musts having a low pH (< or = 3.6), whereas in some musts having a high pH (> or = 3.7), the cell numbers increased during fermentation. During the process of cold soaking, the cell numbers of acetic acid bacteria also increased until inoculation with commercial wine yeast. Gluconobacter oxydans dominated in the fresh must and Acetobacter pasteurianus and A. liquefaciens during fermentation. This study showed that A. liquefaciens and A. hansenii were present in significant numbers, which has not been reported before. PMID- 11930954 TI - Community development and health project: a 5-year (1995-1999) experience in Mozambique, Africa. AB - The Community Development and Health Project (1995-1999) in Mozambique, East Africa, was undertaken to help alleviate the dire circumstances of daily life following years of internal warfare. The project was supported by the International United Methodist Church and was administered by the Mozambique Conference of the United Methodist Church, Chicuque Rural Hospital and Cambine Health Center. The target population, whose daily survival depended on subsistence farming, lived throughout the country, in particular in rural areas, far from health care facilities. The Project Content included the following topics: basic education in personal and environmental hygiene; proper nutrition using locally available food products; methods of agriculture that would ensure the best use of the land for growing nutritious food; economic methods to guarantee a supply of safe drinking water; disease prevention/home treatment; maternal child health; self-esteem issues; and establishing a representatively inclusive community-development committee. Indigenous women, the 'promotoras/os', were selected from their communities, attended a 4-week training session and then returned to their homes as 'Promotoras/os of Community Development and Health'. PMID- 11930955 TI - Hasten slowly: a needs analysis for nurse and village health worker education in East Timor. AB - This article reports on a needs analysis undertaken to determine the educational needs of nurses and health workers in East Timor. The needs analysis, which used a theoretical framework described by Wass (1994), was conducted in both Australia and East Timor. It addressed the current health status of the East Timorese people and the educational requirements of East Timorese nurses and village health workers. Utilizing interviews, field observations and data from the World Health Organization and the United Nations, the following four categories of needs were assessed: felt; expressed; comparative; and normative. The findings document the almost complete destruction of the health infrastructure in East Timor and demonstrate the urgent need for assistance in the re-establishment and enhancement of nursing and primary health care education programmes. A series of recommendations outlining nurse and village health care worker education programmes are proposed. PMID- 11930956 TI - Depleted uranium report from the Health Council of the Netherlands. PMID- 11930957 TI - A survey of x-ray diagnostic services in Delta State, Nigeria (1991-1994). AB - A survey was conducted of x-ray radiological services in Delta State, Nigeria. Information was gathered through questionnaires and interviews from hospitals and clinics as well as from the State Ministry of Health. The data obtained were analysed. Results show that as of 1994 there were 17 x-ray machines installed at 14 different locations within seven local government areas of the state. Results also show that approximately 1% of the population of this state underwent an x ray examination of one form or another. These two findings suggest that the bulk of the population in the state have no access to x-ray diagnostic services. Thirty five kinds of radiological examination were identified to comprise most of the examinations given. Results also show that the background radiation for different x-ray centres was greater than the background radiation for the environment outside the centres. This value, however, was found to be lowerthan the limitrecommended by the International Commission on Radiological Protection. The exposure dose of x-rays to patients was also investigated, and results show that only eight of the centres have a dose chart for the different examinations. This implies that many patients must have being facing the risk of an overdose of x-rays from the other six centres. A regulatory or supervisory body for medical radiation is proposed to regulate and monitor x-ray services and other radiation sources in Delta State. PMID- 11930958 TI - Whether physician-assisted suicide serves a "legitimate medical purpose" under the Drug Enforcement Administration's regulations implementing the Controlled Substances Act. PMID- 11930959 TI - A review of the National Institutes of Health's "Guidelines for Research Using Human Pluripotent Stem Cells". PMID- 11930960 TI - Implementation of a compact, four-stage, scalable optical interconnect for photonic backplane applications. AB - We report on the implementation of a dense 512-beam free-space optical interconnect linking four optoelectronic VLSI chips at the backplane level. The system presented maximizes the positioning tolerances of the components by use of slow f-number (f/16) Gaussian beams and oversized apertures. A beam-clustering scheme whereby a 4 x 4 array of beams is transmitted by each minilens is used to provide a high channel density. A modular approach is used to decrease the number of degrees of freedom in the system and achieve passive alignment of the modules in the final integration phase. A design overview as well as assembly and experimental results are presented. PMID- 11930961 TI - Pharmacological characterisation of arthritis induced by Bothrops jararaca venom in rabbits: a positive cross talk between bradykinin, nitric oxide and prostaglandin E2. AB - BACKGROUND: Our previous results showed that nitric oxide (NO) and bradykinin (BK) mediate the arthritis induced by Bothrops jararaca venom (BjV) in rabbits. In this study, we investigated the contribution of each receptor of BK as well as the inter-relationship between NO and eicosanoids in BjV-induced arthritis. METHODS: The arthritis was induced in rabbits with 16 microg of BjV injected intra-articularly. Prostaglandin E2 (PGE2), thromboxane B2 (TxB2), leukotriene B4 (LTB4) (radioimmunoassay) and nitrite/nitrate concentrations (NO2/NO3) (Griess reaction) were evaluated in the synovial fluid 4 h later. The animals were prior treated with NO synthase inhibitor (L-NAME; 20 mg/kg/day for 14 days), the B2 antagonist of BK (HOE-140) and the B1 antagonist of BK (des-Arg9[Leu8] bradykinin), both at a dose of 0.3mg/kg, 30 min prior to the venom injection. RESULTS: Data show that L-NAME and HOE-140 treatment were equally able to reduce PGE2 and NO2/NO3 levels without interfering with TxB2 and LTB4 production. On the contrary, the B1 antagonist of BK inhibited TxB2 and LTB4 production, and did not alter PGE2 and NO metabolites levels in the inflamed joint. DISCUSSIONS: The results presented clarify the contribution of the kinin system, mainly through the B2 receptor, to the local inflammatory response induced by BjV, as well as its positive interaction with PGE2 and NO production. PMID- 11930963 TI - Need for objective measures to prove clinical outcome. PMID- 11930962 TI - Evidence for local inflammation in complex regional pain syndrome type 1. AB - BACKGROUND: The pathophysiology of complex regional pain syndrome type 1 (CRPS 1) is still a matter of debate. Peripheral afferent, efferent and central mechanisms are supposed. Based on clinical signs and symptoms (e.g. oedema, local temperature changes and chronic pain) local inflammation is suspected. AIM: To determine the involvement of neuropetides, cytokines and eicosanoids as locally formed mediators of inflammation. METHODS: In this study, nine patients with proven CRPS 1 were included. Disease activity and impairment was determined by means of a Visual Analogue Scale, the McGill Pain Questionnaire, the difference in volume and temperature between involved and uninvolved extremities, and the reduction in active range of motion of the involved extremity. Venous blood was sampled from and suction blisters made on the involved and uninvolved extremities for measurement of cytokines interleukin (IL)-6, II-1beta and tumour necrosis factor-alpha (TNF-alpha), the neuropetides NPY and CRGP, and prostaglandin E2 RESULTS: The patients included in this study did have a moderate to serious disease activity and impairment. In plasma, no changes of mediators of inflammation were observed. In blister fluid, however, significantly higher levels of IL-6 and TNF-alpha in the involved extremity were observed in comparison with the uninvolved extremity. CONCLUSIONS: This is the first time that involvement of mediators of inflammation in CRPS 1 has been so clearly and directly demonstrated. This observation opens new approaches for the succesful use and development of immunosuppressives in CRPS 1. PMID- 11930965 TI - Education versus environmental countermeasures. PMID- 11930964 TI - Analysis of putative interactions between potyviral replication proteins and plant retinoblastoma proteins. AB - Sequence comparisons suggest that the RNA-dependent RNA polymerase (NIb) of potyviruses and bymoviruses, as well as the viral polymerase of potexviruses may contain a putative retinoblastoma protein (pRb) binding motif. The possibility that the potyviral NIb may function in the nucleus through interactions with plant pRb-related (RBR) proteins, and the modifications of the cell cycle was investigated by a combination of mutagenesis of the NIb and yeast two-hybrid system (YTHS). Mutation of a highly conserved glutamic acid residue in the putative pRb-binding motif of the NIb had no detectable phenotypic effect on replication of Potato virus A (PVA). Furthermore, the NIb proteins from Potato virus V and PVA failed to interact with maize or tobacco RBR proteins in yeast. Although the conservation of the motif for pRb interaction in plant RNA viruses is intriguing, these proteins from plant RNA viruses appear not to interact with plant RBR proteins. PMID- 11930966 TI - Setting priorities in injury prevention: the application of an incidence based cost model. AB - OBJECTIVES: To make detailed calculations on the direct medical costs of injuries in the Netherlands to support priority setting in prevention. METHODS: A computerised, incidence based model for cost calculations was developed and incidence figures derived from the Dutch Injury Surveillance System (LIS) which provides national estimates of the annual number of patients treated at an emergency department. A comprehensive set of cost elements (that is, health care segments) was obtained from health care registrations and a LIS patient survey. Patients were assigned to specific groups based on LIS characteristics (for example, age, injury type). Average costs per patient group were calculated for each cost element and total costs estimated by adding costs for all patient groups. RESULTS: The direct costs of injury average 2000 guilders per injury patient attending an emergency department. Home and leisure injuries account for over half of the costs, although cost per patient is highest for motor vehicle injuries. Injuries to the lower extremities account for almost half of the total costs and are incurred mainly in the home or recreation. Motor vehicle crashes are the major cause of head injuries. CONCLUSIONS: The model permits continuous and detailed monitoring of injury costs. Estimates can be compiled for any LIS patient group or injury subcategory. The results can be used to rank injuries for prioritisation of prevention by injury categories (for example, traffic, home, or leisure), or by specific scenarios (for example, fall at home). PMID- 11930967 TI - Doolin lecture 2001: Dr. Fenton Howell 'Towards a tobacco free society: should we care? PMID- 11930968 TI - The implementation of the Fast Track program: an example of a large-scale prevention science efficacy trial. AB - In 1990, the Fast Track Project was initiated to evaluate the feasibility and effectiveness of a comprehensive, multicomponent prevention program targeting children at risk for conduct disorders in four demographically diverse American communities (Conduct Problems Prevention Research Group [CPPRG], 1992). Representing a prevention science approach toward community-based preventive intervention, the Fast Track intervention design was based upon the available data base elucidating the epidemiology of risk for conduct disorder and suggesting key causal developmental influences (R. P. Weissberg & M. T. Greenberg, 1998). Critical questions about this approach to prevention center around the extent to which such a science-based program can be effective at (1) engaging community members and stakeholders, (2) maintaining intervention fidelity while responding appropriately to the local norms and needs of communities that vary widely in their demographic and cultural/ethnic composition, and (3) maintaining community engagement in the long-term to support effective and sustainable intervention dissemination. This paper discusses these issues, providing examples from the Fast Track project to illustrate the process of program implementation and the evidence available regarding the success of this science-based program at engaging communities in sustainable and effective ways as partners in prevention programming. PMID- 11930969 TI - Evaluation of the first 3 years of the Fast Track prevention trial with children at high risk for adolescent conduct problems. AB - Fast Track is a conduct-problem prevention trial that derives its intervention from longitudinal research on how serious and chronic adolescent problem behaviors develop. Over 9,000 kindergarten children at 4 sites in 3 cohorts were screened, and 891 were identified as high risk and then randomly assigned to intervention or control groups. Beginning in Grade 1, high-risk children and their parents were asked to participate in a combination of social skills and anger-control training, academic tutoring, parent training, and home visiting. A multiyear universal classroom program was delivered to the core schools attended by these high-risk children. By the end of third grade, 37% of the intervention group was determined to be free of serious conduct-problem dysfunction, in contrast with 27% of the control group. Teacher ratings of conduct problems and official records of use of special education resources gave modest effect-size evidence that the intervention was preventing conduct problem behavior at school. Parent ratings provided additional support for prevention of conduct problems at home. Parenting behavior and children's social cognitive skills that had previously emerged as proximal outcomes at the end of the 1st year of intervention continued to show positive effects of the intervention at the end of third grade. PMID- 11930970 TI - Predictor variables associated with positive Fast Track outcomes at the end of third grade. AB - Progress has been made in understanding the outcome effects of preventive interventions and treatments designed to reduce children's conduct problems. However, limited research has explored the factors that may affect the degree to which an intervention is likely to benefit particular individuals. This study examines selected child, family, and community baseline characteristics that may predict proximal outcomes from the Fast Track intervention. The primary goal of this study was to examine predictors of outcomes after 3 years of intervention participation, at the end of 3rd grade. Three types of proximal outcomes were examined: parent-rated aggression, teacher-rated oppositional-aggressive behavior, and special education involvement. The relation between 11 risk factors and these 3 outcomes was examined, with separate regression analyses for the intervention and control groups. Moderate evidence of prediction of outcome effects was found, although none of the baseline variables were found to predict all 3 outcomes, and different patterns of prediction emerged for home versus school outcomes. PMID- 11930971 TI - Prevention/Intervention trials and developmental theory: commentary on the Fast Track special section. AB - Well-conducted prevention and intervention research has the potential to serve the dual ends of enhancing children's adaptive outcomes and elucidating important developmental processes and mechanisms related to change. Admirably, the Fast Track preventive intervention was conceived in accord with explicit developmental theory related to the development of conduct problems. Herein, I consider how the present reports of grade-3 outcomes allow examination of causal processes and developmental mechanisms related to the effects uncovered, featuring the constructs of prediction, moderation, and particularly mediation. The multifaceted, intensive nature of the Fast Track preventive intervention is at once a clinical strength and a liability in terms of isolating causal processes underlying child change. I also consider issues related to the random assignment of the investigation and to policies that may emerge from the findings. In all, research that aims to identify relevant developmental and causal processes must incorporate both experimental and nonexperimental paradigms that are conceptualized from the outset with explanatory purposes in mind. PMID- 11930972 TI - The Fast Track Project: a seminal intervention efficacy trial. AB - The Fast Track Project is a seminal intervention efficacy trial with several hallmark features, including theoretically driven intervention components, rigorous implementation, and integrity in how results are characterized. Critical issues are discussed in relation to sustainability, a controversy over what really constitutes prevention, and replicability. PMID- 11930973 TI - Intergenerational transmission of risks for problem behavior. AB - The intergenerational transmission of risk factors for problem behaviors was examined across three generations. Two hundred fifty-four 2-year-old toddlers, one or two of their parents, and one grandmother of each toddler were studied. Grandmothers and parents were individually interviewed. Data were analyzed for the male and female toddlers combined. Correlations and hierarchical multiple regression analyses were performed. Findings indicate that the grandmother-parent relationship, parental personality attributes, marital harmony, and drug use and the parent-toddler relationship, predict the toddlers' behavior. The investigation provides evidence for a longitudinal, intergenerational process whereby the grandmother-parent relationship and the parents' personality and behavioral attributes are transmitted across generations through their association with the parent-child relationship. PMID- 11930975 TI - Cognitive response repertoires to child noncompliance by mothers of aggressive boys. AB - Cognitive response repertoires to videotaped child noncompliance episodes were examined in mothers of aggressive (MAs) and nonaggressive 4-6-year-old boys. Mothers provided open-ended solutions to three subtypes of child noncompliance under conditions of time pressure, or after they waited for 15 s to consider alternatives. Solutions were coded as assistance/facilitation, coercion, deference, or explanation/clarification. Compared with controls, MAs offered fewer explanation/clarification responses, more coercive responses, and fewer unique solutions during pressured responding. Two to 6 weeks later, mothers were videotaped while participating with their sons in a challenging block building task. Maternal responses to the vignettes predicted conflict escalation during block building, even after rates of concurrent and past child noncompliance were partialled out. Implications for parent-training models are considered. PMID- 11930974 TI - Responsiveness in interactions of mothers and sons with ADHD: relations to maternal and child characteristics. AB - We observed mother-child interactions, at baseline, in 136 families of 7-10-year old boys with attention-deficit/hyperactivity disorder (ADHD) who were part of a large clinical trial, the Multimodal Treatment Study of Children with ADHD. Independent coders rated stylistic aspects of maternal behavior and factor analyses revealed a responsiveness factor that included overall responsiveness and sensitivity to the child, warmth and acceptance, and appropriate control. We examined relations between maternal responsiveness and (a) maternal depressive symptoms and maternal childhood ADHD symptoms, and (b) boys' ADHD and conduct problem symptoms. Controlling for all other variables, maternal responsiveness displayed a unique negative association with mother-reported child conduct problems, but not with child ADHD symptoms, and also was negatively related to maternal depressive symptoms. We discuss the unique association between mother reported child conduct problems and parenting, and note the utility of studying parenting style in families of children with ADHD. We describe the results within the framework of a transactional model. PMID- 11930976 TI - Intraportal hepatocyte transplantation in the pig: hemodynamic and histopathological study. AB - BACKGROUND: Hepatocyte transplantation is an attractive treatment for various liver diseases. The intraportal route of transplantation is favored, but little information is available on the possible adverse effects in this technique. We investigated the influence of intraportal loads of hepatocytes on portal, pulmonary, and systemic hemodynamics in 13 pigs. METHODS: Under general anesthesia, pigs were provided with an arterial line, a Swan-Ganz catheter, and two intraportal catheters, one for cell infusion and one for heparin infusion and portal pressure measurement. Pig hepatocytes were infused at a rate of 25 million cells/min. RESULTS: The first six animals were used to develop the infusion technique. In the last seven animals, portal pressure increased linearly with cell load upon infusion of 400-2400 x 10(6) hepatocytes (r(2)=0.704;P<0.05). Portal flow measured by Doppler sonography decreased by 23-66% below basal values. An inverse linear relationship was found between portal pressure and portal flow (r(2)=0.679; P<0.05), portal flow approaching zero for portal pressure >40 mmHg. Pulmonary arterial pressure increased by 11-62%. AST increased up to 10-fold, and platelets decreased by 22-58%. Hepatocytes-containing thrombi were present in segmental and in smaller portal branches. Hepatocytes were always identified in lung sinusoids 48 hr after infusion, and a small basal pulmonary infarction was found in one animal. CONCLUSION: . These data suggest that up to 2.4% of total hepatocyte mass can be infused in this large animal model. However, the risk of significant thrombotic complications should be considered for clinical applications. PMID- 11930977 TI - Fracture of an exeter stem 3 years after impaction allografting--a case report. PMID- 11930979 TI - Finding a balance. PMID- 11930978 TI - Applications of Kalman filtering to real-time trace gas concentration measurements. AB - A Kalman filtering technique is applied to the simultaneous detection of NH3 and CO2 with a diode-laser-based sensor operating at 1.53 micrometers. This technique is developed for improving the sensitivity and precision of trace gas concentration levels based on direct overtone laser absorption spectroscopy in the presence of various sensor noise sources. Filter performance is demonstrated to be adaptive to real-time noise and data statistics. Additionally, filter operation is successfully performed with dynamic ranges differing by three orders of magnitude. Details of Kalman filter theory applied to the acquired spectroscopic data are discussed. The effectiveness of this technique is evaluated by performing NH3 and CO2 concentration measurements and utilizing it to monitor varying ammonia and carbon dioxide levels in a bioreactor for water reprocessing, located at the NASA-Johnson Space Center. Results indicate a sensitivity enhancement of six times, in terms of improved minimum detectable absorption by the gas sensor. PMID- 11930980 TI - Transmyocardial revascularization plus coronary artery bypass graft surgery to treat coronary artery disease. AB - This report applies to patients who will have bypass surgery for coronary artery disease and have one or more arteries that cannot be treated with surgery. PMID- 11930981 TI - Culture and body image in Western society. PMID- 11930982 TI - Zinc supplementation in the treatment of anorexia nervosa. AB - The clinical manifestations of zinc deficiency and anorexia nervosa are remarkably similar, and a number of studies have demonstrated that there may be a positive correlation between zinc therapy and the rate of recovery of anorexia nervosa patients. However, because of the different interpretations of the results of these studies, the use of zinc supplementation varies. This article examines the evidence supporting zinc supplementation in the treatment of anorexia nervosa. Randomised, double-blind, controlled clinical trials indicate that zinc therapy enhances the rate of recovery in anorexia nervosa patients by increasing weight gain and improving their levels of anxiety and depression. On the basis of these findings and the low toxicity of zinc, zinc supplementation should be included in the treatment protocol for anorexia nervosa. PMID- 11930983 TI - Effects of restraint stress and serotonin on macronutrient selection: a rat model of stress-induced anorexia. AB - The present study investigated the effects of brief (20 min), acute (2 hr) and chronic restraint stress (2 hr/day for five days) at the time of dark onset on macronutrient selection in female Wistar rats. The role of 5-HT1A receptors in nutrient intake was also examined in a dose-response study (100-700 microg/kg body weight) of 8-hydroxy-2-(di-n-propylamino) tetralin (DPAT) in an acute restraint model. A total of 200 rats aged 6-12 months were tested. In all cases, the restraint stress took the form of confinement within tight-fitting cylinder tubes. In the brief and acute paradigms, the intake of pure carbohydrate (CHO), protein (PRO), and fat was measured for 40 min after a single exposure to stress; in the chronic model, nutrient intake was assessed for 40 min only after the final restraint stress session on day 5. In the DPAT paradigm, intraperitoneal injections of the 5-HT1A agonist were given prior to acute restraint and a 40 min session of nutrient self-selection. Statistical analysis using the t-test for independent samples revealed that neither PRO nor total intake was significantly altered by restraint in the brief, acute or chronic restraint experiments, whereas CHO consumption was suppressed by acute (p=0.02) and chronic restraint (p=0.021), and fat intake was suppressed by brief (p=0.010), acute (p=0.002) and chronic restraint (p=0.001). In the DPAT paradigm, acute restraint stress suppressed CHO (p=0.0001), fat (p=0.000001) and total intake (p=0.003). These effects were not reversed by DPAT, and the administration of 300 pg/kg actually further reduced fat intake. In conclusion, fat intake is more sensitive to the suppressive effects of restraint stress than CHO intake, whereas protein intake is unaffected. Furthermore, 5-HT1A receptors do not seem to play a role in the effects of acute restraint on CHO and fat intake. PMID- 11930984 TI - Early family mealtime experiences and eating attitudes in normal weight, underweight and overweight females. AB - The aim of this study was to examine young women's recollections of family mealtimes in order to determine whether these memories may serve to differentiate those of normal weight from those who were underweight or overweight. A sample of 340 women selected on the basis of their enrollment in a college course completed the Childhood Family Mealtimes Questionnaire and the short form of the Eating Attitudes Test (EAT). The women were divided into under, normal and overweight groups on the basis of their body mass index (BMI). Two factors (appearance weight control and emphasis on mother's weight) were significantly higher among the overweight than the underweight women, and mealtime communication-based stress was highest among the underweight women. The use of family mealtimes as a window for understanding the development of eating disorders (ED) is discussed. PMID- 11930985 TI - Estimation of the population "at risk" for eating disorders in a non-clinical Swedish sample: a repeated measure study. AB - OBJECTIVE: To estimate the late-adolescent population "at risk" for eating disorders (EDs), establish its stability over time using repeated measures, and analyse its social and demographic background factors. METHOD: The Eating Disorder Inventory (EDI) and a number of related questions were administered to students in their last year of high school in five Swedish towns in 1993 (n=3855) and 1998 (n=2925). RESULTS: It was found that 6% of girls were "at risk" for eating disorders with no increase over time. There was an association between this risk and certain socio-demographic factors (living alone, moving away from home early, excessive physical training and experiences of bullying). No differences were found between cities of different size. CONCLUSION: A considerable proportion of girls in late adolescence are preoccupied with weight and dieting, and may be considered "at risk" for developing a clinical eating disorder. This proportion seems to be fairly stable over time. PMID- 11930986 TI - Eating behavior of ballet dancers. AB - Ballet dancers are frequently regarded as having a higher risk of developing eating disorders (ED). This paper describes the eating habits and prevalence of ED in a group of female students from a dance academy in Rome, Italy. Participants were assessed with an array of measures conventionally employed (usually singly) in epidemiological studies of ED, namely: an anthropometrical nutritional evaluation, the EAT, EDI, and BUT questionnaires, and the EDE interview. The 160 students who agreed to participate were evaluated anthropometrically, nutritionally and psychometrically and 83 underwent the EDE structured interview. Their calorie intake was insufficient in all age groups in terms of the nutritional standards required by their daily physical activity. EAT, EDI and BUT enhanced concerns about dieting, food intake control and body image. The significance of the correlations between calorie intake and the EAT Dieting and the EDI Perfectionism and Interceptive Awareness scores increased in function of age. Food, weight and body image concerns increased with age and length of time in the ballet environment The reduced calorie intake was not necessarily linked to the presence of psychopathological signs. PMID- 11930988 TI - Rep-PCR--a variant to RAPD or an independent technique of bacteria genotyping? A comparison of the typing properties of rep-PCR with other recognised methods of genotyping of microorganisms. AB - The paper presents technical aspects of rep-PCR fingerprinting technique and compares its typing abilities, differentiation power and reproducibility with other recognised and recommended genotyping methods. Although rep-PCR fingerprinting is similar to MAAP techniques, it demonstrates some essentially different elements. The data presented in this review, indicate a rep-PCR genomic fingerprinting technique as a highly discriminating, independent screening method for determining the taxonomical diversity of bacterial population. PMID- 11930987 TI - Physical activity does not mitigate G-protein-related genetic risk for obesity in individuals of African descent. AB - The G-protein beta3 subunit 825 TT genotype has been associated with obesity and hypertension. We examined the interaction between the G-protein TT genotype, physical activity and body mass index (BMI) in a cross-sectional study of African immigrants and African Americans. The genotype frequencies were 6.3% CC, 37.7% CT, and 56% TT. After adjusting for potential confounders, BMI was found to be significantly higher in the sedentary than in the physically active participants (p=0.045). There was no statistically significant effect for genotype (p=0.215) or the interaction between genotype and the level of physical activity (p=0.219). However, the individuals with the CC or CT genotype who were physically active had substantially lower BMIs (M+/-SE) (i.e., 25.74+/-2.02) than any of the other groups: sedentary CC + CT (30.58+/-1.03), sedentary TT (30.65+/-1.00) or active TT (29.43+/-1.65). Because of the low statistical power of this study, further research is needed to confirm these findings and to explore potential gene environment/lifestyle interactions. PMID- 11930989 TI - Use of molecular techniques in bioremediation. AB - In a practical sense, biotechnology is concerned with the production of commercial products generated by biological processes. More formally, biotechnology may be defined as "the application of scientific and engineering principles to the processing of material by biological agents to provide goods and services" (Cantor, 2000). From a historical perspective, biotechnology dates back to the time when yeast was first used for beer or wine fermentation, and bacteria were used to make yogurt. In 1972, the birth of recombinant DNA technology moved biotechnology to new heights and led to the establishment of a new industry. Progress in biotechnology has been truly remarkable. Within four years of the discovery of recombinant DNA technology, genetically modified organisms (GMOs) were making human insulin, interferon, and human growth hormone. Now, recombinant DNA technology and its products--GMOs are widely used in environmental biotechnology (Glick and Pasternak, 1988; Cowan, 2000). Bioremediation is one of the most rapidly growing areas of environmental biotechnology. Use of bioremediation for environmental clean up is popular due to low costs and its public acceptability. Indeed, bioremediation stands to benefit greatly and advance even more rapidly with the adoption of molecular techniques developed originally for other areas of biotechnology. The 1990s was the decade of molecular microbial ecology (time of using molecular techniques in environmental biotechnology). Adoption of these molecular techniques made scientists realize that microbial populations in the natural environments are much more diverse than previously thought using traditional culture methods. Using molecular ecological methods, such as direct DNA isolation from environmental samples, denaturing gradient gel electrophoresis (DGGE), PCR methods, nucleic acid hybridization etc., we can now study microbial consortia relevant to pollutant degradation in the environment. These techniques promise to provide a better understanding and better control of environmental biotechnology processes, thus enabling more cost effective and efficient bioremediation of our toxic waste and contaminated environments. PMID- 11930990 TI - A model of structure and action of Sau3AI restriction endonuclease that comprises two MutH-like endonuclease domains within a single polypeptide. AB - Sau3AI is a type II endonuclease that cleaves GATC sequences, producing sticky ends with 4-nucleotide 5'-overhangs. Its activity is inhibited by cytosine C5 methylation within the target sequence. In the N-terminus, Sau3AI exhibits sequence similarity to the GATC-specific single-strand nicking endonuclease MutH implicated in mismatch repair (Ban and Yang, 1998). Sequence analysis of Sau3AI and its homologs reveals that Sau3AI possesses an additional MutH-like domain in the C-terminus. Structure prediction suggests that the C-terminal domain lacks the endonuclease active site but retains all putative DNA-binding elements. As an illustration of these findings, a model of quaternary structure of Sau3AI complexed with the target DNA is presented. These predictions have implications for evolution, structure and function of bacterial DNA repair enzymes and restriction endonucleases. PMID- 11930991 TI - Application of rep-PCR fingerprinting for genotyping of Escherichia coli strains in Wojnowskie Wschodnie and Wojnowskie Zachodnie lake. AB - The paper presents usefulness of application of the PCR-based fingerprinting method, which uses enterobacterial repetitive intergenic consensus primers ERIC (rep-PCR (ERIC)) in analysing and characterising Escherichia coli population in the water environment. 46 E. coli isolates of homogenous biochemical properties were analysed. The received results prove considerable genomic diversity among the analysed isolates. The used technique has turned to be a reproducible and rapid method with a considerable differentiation power. The introductory research has revealed that the technique may be successfully used in qualitative research, for intra-species differentiation of microorganisms occurring in water environment. PMID- 11930992 TI - Contribution of hemolysin and phospholipase activity to cytolytic properties and viability of Listeria monocytogenes. AB - The life cycle of Listeria monocytogenes strongly depends on cytolysin which facilitates the pathogen's escape from the endosome. This process is very ineffective in the absence of phospholipases. In this study we investigated the molecular mechanism of cooperation between hemolysin and phospholipases. We propose a model involving interactions between proteins that increase the affinity of cytolysins for the membrane as well direct enzymatic enhancement of lysis. PMID- 11930993 TI - Characterization of Staphylococcus aureus strains isolated from cystic fibrosis patients by conventional and molecular typing. AB - The phenotypic and genotypic characteristics of Staphylococcus aureus strains isolated from respiratory tract of cystic fibrosis (CF) patients were investigated. Slime production, cell-surface hydrophobicity, type of capsular polysaccharide, profile of heteroresistance to methicillin and Sma I restriction profiles were evaluated. S. aureus CF strains have been shown to be heterogeneous in respect to several important features. All of them were slime producing with variation in colony morphology. High or moderate cell-surface hydrophobicity (CSH) was found for, respectively, 16.2% and 83.8% strains. Thirty strains were resistant to methicillin, 60% of them showed heteroresitance and 40% were homoresistant. It was found that 59.6% of strains produced capsular polysaccharides (CP) of 5 or 8 type. Among CP5/CP8 strains, CP8 was the predominant type (81.1%). Typing of 62 CF strains by macrorestriction analysis of chromosomal DNA revealed several major types, differing in their SmaI profiles with a similarity coefficient lower than 0.4. Some of the strains isolated from the same patient at different times of hospitalization, as well as strains isolated at the same time from the relatives, were identical in their PFGE pattern. PMID- 11930994 TI - Detection of cytomegalovirus in infant cerebrospinal fluid by conventional PCR, nested PCR and PCR-Digene. AB - The possibility of amplification of human cytomegalovirus (HCMV) DNA in cerebrospinal fluid (CSF) for the diagnosis of HCMV central nervous system (CNS) infection in infants was studied. Single-step PCR, nested PCR and PCR-Digene were used to assay CSF specimens from 37 patients. Criteria for patient inclusion in the study were: 1. clinical manifestations suggesting CMV neuroinfection such as seizures, hypertonia, hypotonia, intracranial calcification, microcephaly, chorioretinitis; 2. any of the following symptoms: anaemia, hepetomegaly, prolonged cholestatic jaundice, or hepatitis, splenomegaly, thrombocytopenia, intrauterine hypotrophy; 3. serologic presentation, and/or positive results for CMV infection obtained by single-step PCR and PCR-Digene in urine and/or blood. PCR-Digene results were positive in 6 CSF samples. Four CSF samples were positive by nested PCR and 1 CSF sample by single step PCR. We found that the double PCR was about ten or more times more sensitive than single PCR and the PCR-Digene was only three times more sensitive than nested-PCR. The results were correlated with serology. Thirty-three out of 37 examined patients were seropositive (ELISA IgG); ELISA IgM gave positive results in 9 patients. In control studies, cells infected with other members of the herpes virus family were negative with these methods, which suggest that amplification combined with primers from the IE and the L region of CMV is specific. In conclusion, nested-PCR seems to be the best method for early diagnosis of CMV infection in CSF due to an absence of false positive results and its high specificity and sensitivity. PMID- 11930995 TI - Detection of EBV infection in different etiologic groups of patients. AB - The ability of two diagnostic tests (ELISA and IF) to detect of EBV infection in etiologically different group of patients was compared: cases of chronic lymphoadenopatis, confirmed mononucleosis or suspected EBV infection, tumors like leukemia or lymphoma. The presence of specific IgM and IgG antibodies for different EBV antigens was studied. The results obtained indicated that as many as 17 out of 32 tested serum samples presented different interpretation of EBV infection in both tests used. High number of discordant results was observed in detection of EA-IgG. The highest number of discordant results was observed in group of patients with tumors, while the lowest in group of cases diagnosed or suspected for EBV infection. PMID- 11930996 TI - Biotransformation of phosphogypsum in media containing different forms of nitrogen. AB - Studies on the biotransformation of phosphogypsum (a waste product formed in the course of the production of phosphorous fertilizers) with the use of sulfate reducing bacteria (SRB) demonstrated that it is a good source of sulfates and biogenic elements for these bacteria, though the addition of organic carbon and nitrogen is necessary. The aim of this study was to investigate the form of nitrogen and C:N ratio in the medium on the growth of SRB community in cultures containing phosphogypsum. Batch community cultures of sulfate reducing bacteria were maintained in medium with phosphogypsum (5.0 g/l), different concentrations of sodium lactate (1.6 - 9.4 g/l) and different forms (NH4CI, CO(NH2)2, KNO3) and concentrations (0 - 250 mg/l) of nitrogen. The growth of SRB was studied in the C:N ratio of from 2:1 to 300:1. It was found that: 1 - the best source of nitrogen for SRB is urea, followed by ammonium, the worst were nitrates; 2 - the bacteria were also able to grow in medium without nitrogen but their activity was then by approximately 15% lower than in optimal growth conditions; 3 - in medium with KNO3 inhibition of sulfate reduction by approx. 50% was observed; 4 - the highest reduction of nitrates (removal of nitrate) in media with phosphogypsum and nitrates was at limiting concentrations of sodium lactate. This is probably caused by the selection under these conditions (low concentration of hydrogen sulfide) of denitrifying bacteria or sulfate reducing bacteria capable of using nitrates as an electron acceptor. PMID- 11930997 TI - Application of Saccharomyces cerevisiae and Pichia stipitis karyoductants to the production of ethanol from xylose. AB - Karyoductants of Saccharomyces cerevisiae V30 and Pichia stipitis CCY 39501 with the ability to ferment D-xylose to ethanol were isolated. The ability of these isolates to assimilate different sugars, ethanol tolerance and ethanol production from D-xylose was investigated. Karyoductants didn't grow on starch, lactose and cellobiose, like S. cerevisiae, but showed good growth on xylose and L-arabinose, like P. stipitis. All isolates fermented xylose to ethanol slower than P. stipitis and with lower yields, 0.09 - 0.16 g/g. They secreted also about 3.4 - 7.1 g/dm3 of xylitol to the culture medium (P. stipitis only 0.06 g/dm3). The karyoductants showed an average tolerance to ethanol when compared with the parent strains and fermented glucose in the presence of 6% alcohol whereas parent strain S. cerevisiae and P. stipitis showed exogenic ethanol tolerance of 9% and 3%, respectively. PMID- 11930998 TI - Human herpesvirus type 8 seroprevalence among patients in immunosuppression state. AB - Human herpesvirus 8 (HHV 8) is implicated in the etiology of neoplastic disorders, especially in HIV-infected and immunosuppressed people, but knowledge about seroprevalence of HHV 8 in general or selected populations is still insufficient. In this study the presence of IgG antibodies to HHV 8 in groups of immunosuppressed patients was tested. PMID- 11930999 TI - Production of alpha-amylase and xylanase by an alkalophilic strain of Penicillium griseoroseum RR-99. AB - An alkalophilic strain of Penicillium sp. RR 99 was isolated that was found to synthesise extra-cellular alpha-amylase and xylanase, when cultivated in presence of starch and xylan respectively. The strain showed maximum alpha-amylolytic activity on 4th day and maximum xylanolytic activity on 6th day of cultivation. The ability of the strain to hydrolyse starchy and hemicellulosic wastes made the strain competent not only for the commercial production of these enzymes but also for successful utilization of wastes. PMID- 11931000 TI - Adherence of Pseudomonas aeruginosa strains to solid surfaces. AB - The aim of this study was to evaluate adherence of 59 strains of Pseudomonas aeruginosa to nitric-acid cleansed glass surfaces. There were differences in adherence between the investigated strains. The highest adherence was noticed among human strains (the average percentage was 13.3 +/- 7.51%) and the lowest adherence was determined among swine strains (the average percentage amounted 6 +/- .37%). We conclude that strains of Pseudomonas aeruginosa isolated from humans colonise glass surfaces better than strains isolated from animals. PMID- 11931001 TI - Depressive cognition: a test of depressive realism versus negativity using general knowledge questions. AB - This research is an examination of whether cognition in depressed individuals incorporates a realistic view of the world or a general tendency toward negativity. Participants provided two types of probability judgments of the likelihood that they correctly answered general knowledge questions: the probability that they correctly answered each of the individual questions and an aggregate judgment, after completing the questionnaire, of the percentage of all the questions they thought they had correctly answered. These tasks generally elicit overconfidence and accuracy in nondepressives. In accord with theories of both depressive realism and general negativity, in their item-by-item assessments of their answers to the individual questions, depressed participants demonstrated less overconfidence than nondepressed participants. In accord with the theory of general negativity but not with the theory of depressive realism, however, depressed participants demonstrated underconfidence in their aggregate judgments. The implications of these findings on theories of depressive cognition are discussed. PMID- 11931003 TI - Do thinking styles contribute to academic achievement beyond self-rated abilities? AB - This research identifies individual differences in academic achievement attributable to thinking styles over and above what can be explained by self rated abilities. Participants were 209 university students from Hong Kong and 215 university students from mainland China. Participants responded to the Thinking Styles Inventory (Chinese version) that is based on Sternberg's theory of mental self-government (R. J. Sternberg, 1988). They also rated their own analytical, creative, and practical abilities on a 10-point scale based on R. J. Sternberg's (1985) triarchic theory of human intelligence. Participants' academic achievement scores were also used. The prediction that thinking styles statistically predict academic achievement was supported by data from both Hong Kong and mainland China. Academic achievement and thinking styles are related differently in the two groups. Implications of these findings for both teaching and research are discussed. PMID- 11931002 TI - Deception detection: interrogators' and observers' decoding of consecutive statements. AB - This study is an examination of two forensically important but previously neglected issues in interpersonal deception. First, which cues do lie catchers who have access to repeated interrogations-pay attention to in order to detect deception? Second, do face-to-face interacting interrogators differ from noninteracting observers in terms of how they perceive a suspect? After watching a staged event, 24 suspects (12 liars and 12 truth tellers) were interrogated three times over a period of 11 days. After the final interrogation, the veracity of each suspect was assessed by his or her interrogator and by 6 observers who had watched the interrogations on video only. The results of the experiment showed that consistency over time was by far the most commonly used cue for justifying veracity judgments. Critically, the predictive accuracy for this cue was alarmingly low. As opposed to results from previous research, the interrogators used verbal cues to a significantly greater extent than did the observers. Furthermore, a probing effect was shown (i.e., probed suspects were perceived as significantly more honest than nonprobed suspects). Finally, limited support for a previously reported honesty effect was obtained (i.e., that interrogators perceive suspects to be more honest than do observers). PMID- 11931004 TI - The role of househusband and housewife as perceived by a college population. AB - The research literature frequently conveys the notion that gender roles are becoming less stereotypical. In this study the authors explored the question of how a college population views the role of househusband and housewife and if there are significant differences in role expectations. A 51-item questionnaire was constructed and administered to 526 college students. This sample was chosen because college students are likely to be assuming these roles in the near future. As predicted, the role expectations for househusbands and housewives are not the same. Women tended to question the current structure of household roles more than did men, and the role of househusband was generally more negatively perceived. Reasons for these findings are discussed. PMID- 11931005 TI - The relationship between exercise motives and psychological well-being. AB - The aim of the present study was to use the self-determination model of exercise motives to examine the relationship between extrinsic and intrinsic motives for exercise and a number of measures of psychological well-being. Undergraduate students purporting to exercise regularly (N = 227; 102 men, 125 women) were split into 2 groups: those exercising for less than 6 months and those exercising for 6 months or more. The respondents were asked to complete measures of exercise motivation, self-esteem, psychological well-being, and stress. Among individuals exercising for less than 6 months, a number of extrinsic motivations for exercise were significantly related to poorer psychological well-being. Among individuals exercising for 6 months or more, a number of intrinsic motivations were significantly related to better psychological well-being. The present findings suggest that researchers can use self-determination theory to understand the relationship between exercise motivation and psychological well-being. PMID- 11931006 TI - Antiunion and prounion attitudes as predictors of college students' willingness to join a union. AB - The authors proposed a model of antiunion attitudes and prounion attitudes as predictors of American college students' willingness to join a union (N = 1,245). Structural equation modeling results indicated that antiunion and prounion attitudes are moderately related and both show similar strength in predicting willingness to join. In addition, a general antiunion factor is shown to underlie specific antiunion attitudes, whereas items related to prounion attitudes are shown to reflect a general prounion factor. The utility of the model was supported based on initial and cross-validation samples (ns = 623 and 622, respectively). Implications for measuring union attitudes and recruiting union members among college-educated workers are discussed. PMID- 11931007 TI - A case control study of suicide and attempted suicide in older adults. AB - Risk factors for serious suicidal behavior among older adults were examined in a case control study of 53 adults aged 55 and older who died by suicide or made medically serious suicide attempts and who were compared with 269 randomly selected comparison subjects. Multivariate analyses suggested that risk of serious suicidal behavior was elevated among those with current mood disorders (OR = 179, CI = 52.8-607.6), psychiatric hospital admission within the previous year (OR = 24.4, CI = 1.9-318.7), limited social network (OR = 4.5, CI = 1.4 14.6). The predominant role of mood disorders was confirmed by population attributable risk (PAR) estimates (73.6%), suggesting that the improved detection, treatment and management of mood disorders should be the primary focus of suicide prevention strategies for older adults. PMID- 11931008 TI - Can temperament identify affectively ill patients who engage in lethal or near lethal suicidal behavior? A 14-year prospective study. AB - Among affectively ill patients followed naturalistically for up to 14 years, 36 committed suicide, 120 attempted suicide, and 373 had no recorded suicide attempt. Comparing these three groups on clinical and intake personality revealed that suicide completed within 12 months was predicted by clinical but not personality variables, and suicide beyond 12 months was predicted by newly derived temperament factors, not clinical variables. Attempters and completers shared core characteristics: previous attempts, impulsivity, substance abuse, and psychic turmoil within a cycling/mixed bipolar disorder. Such temperament attributes as impulsivity and assertiveness were the best prospective predictors of completed suicides beyond 12 months with a sensitivity level of 74% and specificity level of 82%. PMID- 11931009 TI - The trajectory of suicidal behavior over time. AB - Some empirical work on genetic and prenatal factors in suicidality is presented. These factors may represent enduring predispositions that comprise risk for initial as well as later suicidal behavior. The existence of enduring predispositions does not preclude the possibility, however, that initial suicidal behavior sets processes into motion that spur later suicidal behavior. Based on past conceptual and empirical work, I suggest two psychological processes- cognitive sensitization and opponent processes--that may partly explain the link between past and future suicidal behavior. PMID- 11931010 TI - Assault victimization and suicidal ideation or behavior within a national sample of U.S. adults. AB - Data from a nationally representative sample of 5,238 U.S. adults were used to examine the extent to which physical assault victimization was associated with suicidal ideation or behavior (SIB). The results from multivariable logistic regression analyses indicate that physical assault victimization was positively associated with SIB after adjusting for sociodemographic characteristics and alcohol use (OR = 3.6; 95% CI = 2.4-5.5). Those who were injured during the most recent physical assault (OR = 2.7; 95% CI = 1.2-6.0) and those who were assaulted by a relative (OR = 3.4; 95% CI= 1.0-11.0) or intimate partner (OR = 7.7; 95% CI = 2.7-22.5) were significantly more like to report SIB than victims who were not injured or were assaulted by a stranger. Also, those who were victimized but not injured (OR = 5.6; 95% CI = 3.8-8.2) and those who were victimized by a stranger (OR = 2.9; 95% CI = 1.4-6.0) were more likely to report SIB than non-victims. These results highlight the need for legal, medical, mental health, and social service providers to address the co-occurrence of violent victimization and suicidal ideation, particularly, but not exclusively, victimization by family members and intimates. PMID- 11931011 TI - Psychiatric inpatients' perceptions of written no-suicide agreements: an exploratory study. AB - One hundred thirty-five psychiatric inpatients admitted for suicidal danger were surveyed regarding their views on the benefits/limitations of written no-suicide agreements. A survey instrument developed for this study revealed that these inpatients, for the most part, rated written no-suicide agreements in a positive manner and in ways consistent with clinical opinion expressed in a number of qualitative/expert-opinion articles. Positive views of no-suicide agreements were not materially influenced by social desirability or age, nor were they moderated by gender, presence/absence of Axis II disorders, or admission suicidal danger. However, patient suicide attempt history (no attempts, one attempt, or more than one attempt) exerted a moderating effect on patients' ratings of the helpfulness of these contracts. Multiple attempters viewed written no-suicide agreements as less helpful than those patients with a single or no prior attempts. The methodological problems and generalizability concerns associated with these results are discussed and future research needs are suggested. PMID- 11931012 TI - Prevention of youth suicide: how well informed are the potential gatekeepers of adolescents in distress? AB - Australia has one of the higher rates of suicide among young people. Although a role for doctors and teachers in the prevention of youth suicide has been suggested, no prior Australian study has assessed adequately the level of suicide knowledge held by these professionals. Knowledge about adolescent suicide was investigated using the Adolescent Suicide Behaviour Questionnaire, a 39-item instrument developed for the purpose. The stratified random sample comprised 404 general practitioners and 481 teachers from 56 secondary schools. General practitioners and teachers scored, on average, 71% and 59% of the questionnaire items correct, respectively. There was wide individual variability: The number of items correct ranged from 4 to 38 for general practitioners and 0 to 34 for teachers. Strengths and deficits in knowledge across the two professions are discussed in terms of the potential gatekeeping role of these professionals in suicide prevention. PMID- 11931013 TI - Suicidal ideation and somatic symptoms of patients with mind/body distress in a Japanese psychosomatic clinic. AB - To examine associations of suicidal ideation with somatic symptoms, 863 outpatients were studied in a psychosomatic clinic in Japan. All subjects were diagnosed according to the third and fourth editions of the Diagnostic and Statistical Manual of Mental Disorders (DSM-III-R and DSM-IV). They completed the Cornell Medical Index Questionnaire to assess suicidal ideation and 15 major somatic symptoms. Mood states were rated using the Profile of Mood States (POMS). In the study, 266 patients (31%) reported suicidal ideation. The age- and sex adjusted odds ratios (typically in the range of 1.5 to 2.5) of somatic symptoms for suicide ideation showed significant differences (all ps < .05) for 13 of 15 symptoms. The total number of somatic symptoms predicted suicidal ideation (p < .05), controlling for the significant effects of sex, education, marital status, and the POMS Depression scale scores through multiple regression analysis. Evaluation of somatic symptoms might be important to assess suicidal ideation in a psychosomatic medicine population. PMID- 11931014 TI - Mass suicide: historical and psychodynamic considerations. AB - Mass suicide can be defined as the simultaneous suicide of all the members of a social group and is closely linked to the human dimension of existence, although the social and cultural context may vary. In fact, the term mass suicide can also be used to describe situations in which a particular population has reacted to oppression by denying all normal activities of sustenance, with the intention of bringing about a traumatic metamorphosis in a cultural context (colonization, exploitation by other populations), thus transforming a catastrophe in which a passive role is played into one constructed actively. Therefore, mass suicides can be subdivided into two categories: (a) hetero-induced, typical of defeated and colonized populations forced to escape from a reality that does not acknowledge their human dignity and (b) self-induced, in which the motivation is related to a distorted evaluation of reality, without there being either an intolerable situation or a real risk of death. The mass suicides that have taken place in the last 20 years are all related to the establishment of religious sects; the mystic delirium created within the sect leads to the self-destruction of the group as being interpreted as an act of self-assertiveness. PMID- 11931015 TI - Reactive oxygen species (ROS) production by amoebocytes of Asterias rubens (Echinodermata). AB - An adapted peroxidase, luminol-enhanced chemiluminescence method in an EDTA-free, Ca++-containing medium is described and used to characterise reactive oxygen species (ROS) production by starfish immunocytes using a standard microplate reader luminometer. ROS production was stimulated by direct interaction of immunocytes with bacteria or bacterial wall components, but not by the soluble stimulant PMA nor the lectin concanavalin A. Produced ROS detected by this method are apparently superoxide anions, hydrogen peroxide and peroxynitrite. Comparison with other chemiluminescence methods indicates that the described method is the only one to detect the stimulation of starfish immunocytes by the Gram-positive bacteria, Micrococcus luteus, a fact that questions previous reports indicating a lack of stimulation by pathogens. The adapted method provides a rapid determination of the overall ROS production, which is suitable for both disease control and immunotoxicological studies in echinoderms. PMID- 11931016 TI - Production of superoxide anion by head-kidney leucocytes of Indian major carps immunised with bacterins of Aeromonas hydrophila. AB - On contact with micro-organisms or any other antigenic particles which are recognised as phagocytosable, the leucocytes of vertebrates raise their oxygen consumption suddenly (respiratory burst). The consumed oxygen is then converted into reactive oxygen species (ROS). An effort has been made in the present study to estimate the production of superoxide anion, one of the ROS, by the resident as well as activated head-kidney leucocytes of Indian major carps. Activation was accomplished by immunisation with formalin killed (FK) and heat killed (HK) whole cell bacterins of Aeromonas hydrophila. In the ex vivo experiment conducted, activated leucocytes yielded a significant increase in O.D. values for nitroblue tetrazolium (NBT) reduction reaction, reflecting an increase in superoxide anion production, from day 1 post immunisation. Of the three species of Indian major carps immunised, Catla catla showed the greatest production of superoxide anion, followed by Labeo rohita and then by Cirrhinus mrigala. The enhancement of superoxide anion production of leucocytes by immunisation justifies the role of immunisation in the microbicidal defence mechanism of fish. PMID- 11931017 TI - The carbohydrate moiety of serum IgM from Atlantic cod (Gadus morhua L.). AB - The carbohydrate moiety of cod serum IgM was analysed using oligosaccharide sequencing techniques. The carbohydrate moiety constituted about 10% of the molecular weight of cod IgM, was associated with the constant region of the heavy chains (Fc), and was composed of N-linked complex type oligosaccharides. Considerable heterogeneity was observed. Sixteen different glycan structures were identified, over 60% were sialylated and 40% contained core fucose. The carbohydrate moiety of cod IgM was shown to provide protection against protease digestion, and partial deglycosylation abolished the antigen binding property of natural cod anti-TNP-BSA antibody. PMID- 11931018 TI - The acute phase response of rainbow trout (Oncorhynchus mykiss) plasma proteins to viral, bacterial and fungal inflammatory agents. AB - The innate arm of the immune system responds to inflammatory stimuli by the activation of phagocytes, and by altered levels of several plasma proteins. These changes in plasma proteins comprise a major component of the acute phase response, which is thought to be an adaptive response that contributes to regaining homeostasis after tissue injury or infection. In this study, rainbow trout (Oncorhynchus mykiss) were injected with a variety of potential inflammatory agents, and changes in the concentrations of plasma proteins were sought in polyacrylamide gels in which plasma proteins had been electrophoresed. Bacteria, viruses and yeast all induced changes in plasma protein profiles. Increases were first evident 2 days after injections, and most were evident within 1 week. The greatest number of changes occurred after injection with a Vibrio bacterin emulsified in Freund's incomplete adjuvant. While some proteins increased and others decreased following several treatments, other proteins changed only in response to injections of viruses or viral proteins, and others changed in response to bacterial components. Some proteins that increased after yeast injection decreased after injection of viral components. The partial amino acid sequence of one increased protein identified it as haptoglobin. PMID- 11931019 TI - Rainbow trout neutrophils are responsible for non-specific cytotoxicity. AB - This is the first report that rainbow trout (Oncorhynchus mykiss) neutrophils are responsible for non-specific cytotoxicity. A monoclonal antibody (mab) for rainbow trout leucocytes was produced. Using this mab (TTL-5E9), neutrophils (5E9 positive cells) were isolated from the pronephros by a panning technique. The isolated neutrophils showed high viability (approximately 95%) and purity (92 95%), and were functional in cytotoxic activity assays. The neutrophils demonstrated significantly higher cytotoxic activities against YAC-1 target cells than the other cells (5E9-negative cells, predominantly lymphocytes). The number of neutrophils contaminating the 5E9-negative fraction and their non-specific cytotoxicities were positively correlated. These findings demonstrate that rainbow trout neutrophils possess non-specific cytotoxic activities. PMID- 11931020 TI - The role of the haematopoietic tissue in haemocyte production and maturation in the black tiger shrimp (Penaeus monodon). AB - The haematopoietic tissue (HPT) of the black tiger shrimp (Penaeus monodon) is located in different areas in the cephalothorax, mainly at the dorsal side of the stomach and in the onset of the maxillipeds and, to a lesser extent, towards the antennal gland. In young and in experimentally stimulated animals, the HPT is expanded in relatively larger and more numerous lobules throughout the cephalothorax. Four cell types could be identified in the HPT by electron microscopy. The type 1 cells are the presumed precursor cells that give rise to a large- and a small-granular young haemocyte, denominated as the type 2 and type 3 cells, respectively. A gradient of maturation from the type 1 towards the type 2 or 3 cells could frequently be observed. The presumed precursor cells are located towards the exterior of the lobules and maturing young haemocytes towards the inner part, where they can be released into the haemal lacunae. The type 4 cells show typical features of interstitial cells. Different stimulation experiments were carried out and various techniques were used to study the HPT in relation to the (circulating) haemocytes. The majority of the cells in the HPT are able to proliferate and proliferation can be increased significantly after the injection of saline and, to a much higher extent, after LPS injection. The circulating haemocytes of crustaceans are generally divided into hyaline (H), semigranular (SG) or granular (G) cells, of which large- and small-granular variants of each of these were suggested in the present study. Even after stimulation in this study, the circulating haemocytes scarcely divide. The high variations that were found in the total haemocyte count in the stimulation experiments were not accompanied by significant differences in differential haemocyte count and, therefore, appeared to be a less useful indicator of stress or health in P. monodon. Light and electron microscopical observations support the regulation of the populations of the different haemocyte types in the circulation by (stored) haemocytes from the connective tissue. In conclusion, according to morphological and immuno-chemical criteria, it is proposed in the present study to divide the haemocytes into a large-and a small-granular developmental series. After extensive morphological observations, it is suggested that the hyaline cells are the young and immature haemocytes of both the large- and small-granular cell line that are produced in the HPT, and can be released into the haemolymph. Indications were found that the granular cells, of at least the large-granular cell line, mature and accumulate in the connective tissue and are easily released into the haemolymph. Combining the results of the present study with literature, this proposed model for haemocyte proliferation, maturation and reaction will be discussed. PMID- 11931021 TI - Production and characterisation of monoclonal anti-idiotype antibody to Vibrio anguillarum. AB - Seven monoclonal anti-idiotype antibodies (mab2) were raised against mouse monoclonal antibody (mab1) 4A6. Identification of subclass showed that 1H5, 1D1, 2B12 and 2F12 belonged to IgG2b, 2H12 and 1H12 to IgG2a and lE10 to IgG3. The titres of these mab2 ascitic fluids ranged from 1 x 10(-4)-1 x 10(-6). The capacity of the mab2 to inhibit the binding between the corresponding rabbit antiserum and Vibrio anguillarum was investigated with the competitive inhibition ELISA. The results showed that mab2 1D1, 1E10, 1H5 and 1H12 were able to inhibit this binding. Another experiment demonstrated that mab2 1D1, 1E10 and 1H5 might induce Balb/c mice to produce Ab3 and these Ab3 competed the same antigen epitopes with Ab1. These results indicate that mab2 1D1, 1E10 and 1H5 are likely to represent an internal image of V. anguillarum and may thus be described as Ab2 beta anti-idiotype antibodies. In protection experiments, Japanese flounders vaccinated with mab21D1, 1E10 and 1H5 showed significantly enhanced survival from challenge with V. anguillarum. Thus. mab21D1, 1E10 and 1H5 may have use as idiotype vaccines for fish in aquaculture. PMID- 11931022 TI - Efficacy of different administration routes for vaccination against Vibrio anguillarum in Atlantic halibut (Hippoglossus hippoglossus L.). AB - Atlantic halibut (Hippoglossus hippoglossus L.) is a potentially important new species to cold-water aquaculture. Development of a viable industrial farming technique has been hampered by continued pathogen problems within the rearing cycle and there are several reports that indicated how susceptible juvenile halibut are to bacterial and viral diseases. Interest has been expressed, within the industry, over the possibility of vaccinating suitably sized animals to protect against the more common aquaculture pathogens. Vibrio spp. are of particular concern due to their ubiquitous nature and the relatively frequent occurrence of these pathogens within marine aquaculture. We have previously investigated the susceptibility of Atlantic halibut to infection by Vibrio anguillarum and the efficacy of intraperitoneal injected delivery of a commercial vaccine in protecting against the disease. Given the very high rate of protection offered by immunisation we wanted to investigate the effect of alternate routes of administration on the efficacy of the vaccine. PMID- 11931023 TI - Prevalence of the agent of human granulocytic ehrlichiosis in Ixodes scapularis (Acari: Ixodidae) in the coastal southeastern United States. AB - Human granulocytic ehrlichiosis (HGE) is an emerging tick-borne disease recently recognized in the United States. The blacklegged tick, Ixodes scapularis Say, is the principle vector in the eastern United States. The disease has been commonly reported in the northeastern and upper midwestern states; however, suitable vectors and reservoir hosts exist in the southeast. To assay the prevalence of the HGE agent in vector ticks, we screened 818 individual I. scapularis from 15 locations in South Carolina, Georgia, and Florida using nested polymerase chain reaction, which targets the HGE agent 16S rRNA gene. Prevalence among locations ranged from 0 to 20%. The overall average prevalence of 15 sites was 1.6% (n = 818). Verification by sequencing the 16S rDNA from the positive samples showed 99.8-100% nucleotide identities with the sequences of the HGE agent in GenBank. These results were supported by the phylogenetic analysis using 16S rDNA sequences. PMID- 11931024 TI - A reinvestigation of brindley's gland exocrine compounds of Rhodnius prolixus (Hemiptera: Reduviidae). AB - A chemical, electrophysiological and behavioral study was carried out to analyze the volatile compounds of the Brindley's gland of Rhodnius prolixus Stal. Six compounds were found in the Brindley's gland of males and females: acetic, isobutyric, caproic acids and three compounds not identified. The mass spectra of the unidentified compounds have the appearance of a propionate ester, a butyrate ester and a valerate ester. There was no difference in the concentration of these compounds in the glands of males and females. The major component was isobutyric acid. The electroantennographic (EAG) evaluation of isobutyric, acetic, and caproic acids elicited weak responses. With both sexes, the EAG responses for the acids were no different from the control response at any of the doses evaluated (1, 10, 100, and 500 microg). The behavioral response of males and females to acetic, isobutyric, and caproic acids at 1, 5, and 10 microg and binary and tertiary mixtures of the three compounds was evaluated in a Y-olfactometer. Males showed preference for acetic acid at a dose of 1 microg, but not at 5 or 10 microg. Females showed preference for this compound at the dose of 5 microg. Males showed preference for isobutyric acid at 1 and 5 microg, but not at 10 microg. Females did not show any preference for isobutyric acid. Males were attracted to caproic acid at the dose of 1 and 10 microg. Females were attracted to the caproic acid. Males, but not females, preferred the mixture of acetic and isobutyric acids over control. Neither males nor females showed preference for the mixture of acetic and caproic acids or hexane control. Females, but not males, were attracted to the mixture of the isobutyric and caproic acids. Only males showed a positive response for the tertiary mixture of the acids. PMID- 11931026 TI - A new species of Polylopadium Brennan & Jones, 1961 (Acari: Trombiculidae) from cricetid rodents, Peru with description of a new character for the genus and a key to the species. AB - peruensis is described as new from specimens collected from Oryzomys yunganus, Proechimys quadruplicatus, and Oryzomys megacephalus taken in Peru. A new character of a nude tactile seta on femur III is noted for the genus and a key to the species of Polylopadium is presented. PMID- 11931025 TI - Immunolocalization of prophenoloxidase in the process of wound healing in the mosquito Armigeres subalbatus (Diptera: Culicidae). AB - Hemolymph coagulation began almost immediately after wounding in mosquito, Armigeres subalbatus, (Coquillett) larvae. Immunocytochemical localization showed that prophenoloxidase (pro-PO) was distributed in the wound site. In the initial wounding, coagulation and wound plug formation occurred with granulocyte migration. The hemocytes lysed and released granular materials around the wound site, prophenoloxidase being mostly localized in granules and cuticle. In the second phase of wound healing, melanin accumulation occurred at the wound site along the margin of the cuticle and rapidly increased in thickness. Immunogold labeled pro-PO was localized in vacuoles, melanins, and cuticle, with the gold particles labeled intensely on the undarkened cuticle and weakly on the darkened cuticle. It is believed that pro-PO is activated upon wound initiation to produce melanin product and deposited on the cuticle. In the final phase of healing, scab melanization and pro-PO immunogold localization were reduced and accompanied by epithelial cell regeneration. This proenzyme was localized in vesicles and flocculent materials, but was absent in the melanized scab. Our results further indicate that pro-PO was present in granules, cuticles, epithelial cells, vacuoles, and flocculent materials but not in melanized scab and coagulated clot. The pro-PO immunogold particles labeled intensely in the initial wounding but weakly in the final phase. Our observations also suggest that pro-PO is released from granulocytes by cell rupture, synthesized or stored in granulocytes, and then is released into the wound site via the cytoplasmic granules. This study indicates that the pro-PO is involved in numerous physiological roles in the process of wound healing in this mosquito. PMID- 11931027 TI - In vitro generation of organophosphate resistant boophilus microplus (Acari: Ixodidae) cell lines. AB - Three organophosphate resistant Boophilus microplus Canestrini cell lines were generated by exposing B. microplus VIII-SCC cell line to incrementally increased toxic concentrations of the acaricide coumaphos. The development of resistance was evidenced by LC50 values elevated over those of control cells. The resistant cell lines selected in higher concentrations of organophosphate, designated C44 and C54, also had significantly slower duplication rates than a resistant cell line selected in lower concentrations of coumaphos (C34) and the nonresistant control cells. Resistant cell lines C44 and C54 also had significantly higher levels of esterase after exposure to coumaphos than resistant cell line C34 and the nonresistant controls. These in vitro results agree with reports of increased esterase activity associated with organophosphate resistance in B. microplus ticks in vivo. PMID- 11931029 TI - Cytogenetics of laboratory colonies of simulium vittatum cytospecies IS-7 (Diptera: Simuliidae). AB - Laboratory colonies of the black fly Simulium vittatum Zetterstedt cytospecies IS 7 were analyzed cytogenetically and compared with the original New York population from which source material was collected 18 yr earlier. All sex chromosomes and major autosomal polymorphisms that were present in the source population were still represented in the laboratory colonies. However, the extent of sex linkage and the frequencies of four of the five major autosomal inversions changed significantly in at least one colony, perhaps because of bottlenecks experienced by the colony. A complete absence of males homozygous for the IS-7 inversion in both field and colony material is explained by postulating that the Y2 chromosome, representing the inverted condition for the IS-7 sequence, is absent from the population or acts as a rare reproductive lethal. This example possibly represents pseudo-partial sex linkage involving the X-linked sequences. PMID- 11931028 TI - Glass capillary tube feeding: a method for infecting nymphal Ixodes scapularis (Acari: Ixodidae) with the lyme disease spirochete Borrelia burgdorferi. AB - We evaluated an artificial capillary feeding method to infect nymphal Ixodes scapularis (Say) ticks with Borrelia burgdoeferi, the causative agent of Lyme disease. Thirty to 70% of the nymphs were infected after feeding for 2.5 h from glass capillary tubes filled with a solution of spirochetes. Capillary infection was stable and persisted in the nymphs for at least 10 d after feeding. Capillary feeding also maintained natural vector competence patterns because I. scapularis ticks acquired infection unlike Dermacentor variablis (Say), which did not become infected. Capillary infected I. scapularis nymphs were capable of transmitting the infection to naive mice although not as efficiently as naturally infected nymphs. The capillary infection method is convenient and is a better alternative to syringe inoculation as a means of infecting animals with B. burgdorferi. PMID- 11931030 TI - Two new species of the genus Periglischrus (Acari: Mesostigmata: spinturnicidae) on two bat species of the genus tonatia (Chiroptera: Phyllostomidae) from southeastern Mexico, with additional data from Panama. AB - Two new species of spinturnicid mites of the genus Periglischrus are described and illustrated from phyllostomid bats from southeastern Mexico: The female, male, and protonymph of Periglischrus steresotrichus, new species, from Tonatia evotis Davis & Carter, and the female and male of Periglischrus eurysternus, new species, from Tonatia saurophila Koopman & Williams. A supplementary description of the male deutonymph of P. eurysternus from T. saurophila from Panama is given. The morphological features of the two new species of Periglischrus are used as a basis for discussing their phylogeny and its potential relationship to that of their hosts. PMID- 11931031 TI - Detection of encephalitis viruses in mosquitoes (Diptera: Culicidae) and avian tissues. AB - ABSTRACT Diagnostic assays for the detection of St. Louis encephalitis (SLE) and western equine encephalomyelitis (WEE) viruses in mosquito pools and avian tissues were compared for sensitivity, accuracy and specificity. The in situ enzyme immunoassay (EIA), plaque assay on Vero cells, passage in Aedes albopictus Skuse C6/36 and C7/10 cells, antigen capture enzyme immunoassay (AC-EIA), and single and multiplex reverse transcription-polymerase chain reactions (RT-PCR) were evaluated using pools of 50 mosquitoes containing 1-2 experimentally infected individuals. RT-PCR was the most sensitive assay, with a detection limit of <0.1 plaque forming unit. AC-EIA was the fastest and most economical procedure, but was the least sensitive, detecting only 38% of positive pools. The in situ EIA included initial virus amplification on Vero cells, thereby improving assay sensitivity to detect 68% of positive pools. Passage in C6/36 and/or C7/10 cell culture revealed the presence of infectious virus in samples positive by RT PCR, but initially negative by plaque assay on Vero cell culture, indicating that detection was related to assay sensitivity and not to the absence of intact infectious virus. Combining WEE and SLE RT-PCR assays into a multiplex assay reduced sensitivity, but stilldetected viral RNA at titers below plaque assay sensitivity. Plaque assay on Vero cells, mosquito cell passage, and several RT PCR procedures were evaluated for their ability to detect WEE and SLE in white crowned sparrow tissues during acute and chronic stages of infection. All assays detected virus during acute infection at times of high viremia; however, only RT PCR assays were positive by day 7 when virus was not detected in sera. RT-PCR detected SLE RNA in spleen tissue from one bird 51 d after infection. Assay sensitivity also was compared using extracts of homogenized bird organs spiked with known titers of WEE and SLE. Trizol RNA extraction followed by Qiagen one step RT-PCR was the most sensitive method, but occasionally resulted in the presence of secondary bands confounding interpretation and requiring confirmatory assays. A balanced surveillance program should combine systems that allow the detection of new agents and the sensitive monitoring of endemic agents to provide an early warning of pending health risks. PMID- 11931032 TI - Evaluation of a dipstick malaria sporozoite panel assay for detection of naturally infected mosquitoes. AB - The determination of the presence or absence of malaria sporozoites in wild caught Anopheles mosquitoes remains an integral component to the understanding of the transmission dynamics in endemic areas. To improve that capability, there has been on-going development of a new device using dipstick immunochromatographic technology for simplifying the testing procedure and reducing the time required to obtain results. As part of a larger multi-center effort, we evaluated the sensitivity and specificity of a prototype malaria sporozoite antigen panel assay (Medical Analysis Systems, Camarillo, CA) against three human Plasmodium species/polymorphs. The wicking (dipstick) assay was compared against a standard parasite antigen capture enzyme-linked immunosorbent assay (ELISA) for the detection of human circumsporozoite protein (CSP) in wild-caught mosquitoes. Over 6,800 Anopheles mosquitoes, representing 20 species collected from malaria endemic areas of Indonesia were tested either individually or in pools of up to 10 mosquitoes each. From 1,442 pooled test strip assays and ELISA formats, nine mosquito pools were found reactive for P.falciparum, P. vivax 210, or P. vivax 247 CSP. There was complete concordance between test strip results and ELISA results. Sensitivity was 100% and given some minor problems with false positives or negatives, specificity (n = 488) was 97%. Most strips judged as false positive produced very weak signals compared with negative control blank strips and paired ELISA-negative samples. The dipstick test proved technically simpler to perform and interpret than the ELISA and results were obtained within 15 min of exposure to mosquito suspension. This qualitative assay appears an attractive alternative to the CSP ELISA for detection of sporozoites in fresh or dried mosquitoes. PMID- 11931034 TI - A new species of the genus Eudusbabekia (Acari: Prostigmata: myobiidae) on Leptonycteris nivalis (Chiroptera: Phyllostomidae) in central Mexico. AB - Eudusbabekia provirilia new species was found on the bat Leptonycteris nivalis (Saussure) in the central part of Mexico. The female, male, protonymph, and larva are described and illustrated. PMID- 11931033 TI - Characterization of the AeaHP gene and its expression in the mosquito Aedes aegypti (Diptera: Culicidae). AB - The sequence and tissue expression of the gene encoding a peptide hormone Aea-HP I, known to inhibit host-seeking behavior, has been characterized for the yellowfever mosquito, Aedes aegypti (L.). The open reading frame reveals a prepropeptide that would be processed into three identical peptides. The gene contains four short introns and exists as a single genomic copy. Transcripts of the gene were present in the brain, terminal ganglion, and midgut of adults, and in females, its expression profile differed for each tissue before and during a reproductive cycle. Peptides resulting from this expression were identified in the female tissues by immunoassays. Numerous neurosecretory cells and neurons in the nervous system were immunostained by an Aea-HP-I antiserum. Hundreds of endocrine cells were stained similarly in the midgut, thus contributing to the 10 times greater amount of immunoreactive peptide in an abdomen than in a head, as determined with an Aea-HP-I radioimmunoassay. Based on these results, neurosecretory cells and midgut endocrine cells are likely sources of Aea-HPs shown to reach highest hemolymph titer at the same time as host seeking is inhibited in female Ae. aegypti during a reproductive cycle. PMID- 11931036 TI - Winter intervention against Aedes aegypti (Diptera: Culicidae) larvae in subterranean habitats slows surface recolonization in summer. AB - At semiarid Charters Towers, north Queensland, Australia, the importance of Aedes aegypti (L.) in wells was assessed in relation to the colonization of surface habitats during the wet season. From April to July 1999, 10 wells (five positive for Ae. aegypti) were monitored to assess their status and larvae population numbers therein. All surface containers located within a 100 m radius of each well were removed, treated with s-methoprene or sealed to prevent the utilization of these containers by mosquitoes. These inner cores were surrounded by outer zones for a further 100 m in which surface containers were left untreated but all subterranean habitats were treated. Ovitraps were monitored monthly in the inner cores for 36 wk from August 1999 to April 2000 and differences in the proportions of ovitraps positive for Ae. aegypti and Ochlerotatus notoscriptus (Skuse) were analyzed by logistic regression. Analysis of the proportions of ovitraps positive for Ae. aegypti near positive wells indicated significantly greater colonization from November to March (the wet season), compared with those situated near Ae. aegypti negative wells. As Oc. notoscriptus were not produced from subterranean sites, comparisons of the proportions of ovitraps positive for Oc. notoscriptus in positive and negative inner cores provided an indication of the relative productivity of the uncontrolled surface containers in the outer zones. Differences in the utililization of ovitraps by Oc. notoscriptus among positive and negative cores were observed during only one month (March), when oviposition was greater in ovitraps in the negative cores, compared with the positive cores. Best subsets linear regression analysis of the proportion of ovitraps positive forAe. aegypti against meteorological variables (rainfall, mean wind speed, mean relative humidity, mean minimum, and maximum temperature) during the week of ovitrapping indicated that minimum temperature and wind speed accounted for 63.4% of the variability. This study confirms that for semiarid towns such as Charters Towers, the practice of treating a relatively small number of key subterranean habitats during winter will significantly affect Ae. aegypti recolonization of surface container habitats during summer, the period of greatest risk for dengue. PMID- 11931035 TI - High malaria transmission intensity in a village close to Yaounde, the capital city of Cameroon. AB - A 2-yr longitudinal malaria study was undertaken in a suburb of Yaounde, the capital city of Cameroon, in the village of Simbock, approximately 2 km from the city limits. This study allowed assessment of malaria transmission intensity and dynamics in this region before implementation of pyrethroid impregnated bed nets through the national vector control program. Anophelines were captured on human volunteers by pyrethrum spray collections and in resting sites outdoors. Malaria vectors were Anopheles funestus Giles, Anopheles gambiae s.s. Giles (M and S forms), Anopheles moucheti Evans, and Anopheles nili Theobald. An. moucheti was the most abundant mosquito captured during the study, accounting for >54% of total anophelines caught. The annual Plasmodium falciparum Welch entomological inoculation rates measured by enzyme-linked immunosorbent assay were 277 infected bites per human for the first year and 368 for the second year. An. gambiae s.s., An. funestus, An. moucheti, and An. nili were responsible for 23.8%, 26.8%, 39.2%, and 10.2% of malaria transmission, respectively. Malaria transmission is perennial throughout the year. All these vectors were highly anthropophagous because only two out of 566 mosquitoes blood-meal tested were not taken on humans. PMID- 11931039 TI - Resistance of the tick dermacentor variabilis (Acari: Ixodidae) following challenge with the bacterium Escherichia coli (Enterobacteriales: Enterobacteriaceae). AB - In addition to a soluble response, many invertebrates control bacterial infections by means of phagocytosis or melanotic encapsulation. In some insects, Escherichia coli growth is reported to be inhibited by aggregation/encapsulation. Soluble and phagocytic responses to bacterial challenge have been reported in ticks, but evidence of an aggregation / encapsulation response was reported only for inanimate (araldite) implants. Ticks were challenged by direct inoculation of bacteria into the hemocoel cavity. By plating, no viable E. coli were detected 6 h postinoculation. A direct fluorescence assay (DFA) revealed aggregated bacteria 1 h postinoculation. Furthermore, DFA showed aggregated bacteria at 6,24, and 48 h postinoculation associated with masses of tissue, presumably of cellular origin, suggesting events similar to those described as nodulation. These findings suggest that encapsulation/ nodulation may be an important component of the immune response in ticks. PMID- 11931038 TI - Effect of diet composition on weight gain, sperm transfer, and insemination in the cat flea (Siphonaptera: Pulicidae). AB - Weight gain by adult cat fleas, Ctenocephalidesfelis (Bouche), was influenced primarily by the concentrations of protein and sodium chloride in the feeding solution. After 48 h of feeding, fleas fed whole blood weighed almost twice as much as fleas fed plasma or hemolyzed blood and 1.25 times as much as fleas fed 0.15 M sodium chloride. When fleas were fed sodium chloride solutions ranging from 0 to 0.5 M, weight gain was greatest on the 0.15- or 0.2-M solutions. Weight gain decreased significantly when fleas were fed plasma, hemolyzed blood or 0.3 or 0.5 M sodium chloride in place of whole blood, but improved when plasma was diluted 100% and when hemolyzed blood was diluted 10% with distilled water. Adenosine-5'-triphosphate did not appear to stimulate weight gain in cat fleas; weight gain was unchanged in fleas fed hemolyzed blood or 0.15 M sodium chloride to which 0.005 M ATP was added. Insemination did not occur in starved fleas or those fed protein-free diets. When fleas were starved or fed distilled water, sodium chloride, or other salt solutions, sperm was transferred from the testes to the vas deferens in 91-94% of males, but no females were inseminated. In contrast, when fleas were fed whole blood, hemolyzed blood, plasma, or bovine serum albumin (3.5 or 7.0 g/deciliter) dissolved in 0.15 M saline, 80, 80, 10, and 10% of the females were inseminated, respectively. PMID- 11931037 TI - Development and evaluation of a species diagnostic polymerase chain reaction restriction fragment-length polymorphism procedure for cryptic members of the Culex sitiens (Diptera: Culicidae) subgroup in Australia and the southwest Pacific. AB - Members of the Culex sitiens subgroup are important vectors of arboviruses, including Japanese encephalitis virus, Murray Valley encephalitis virus and Ross River virus. Of the eight described species, Cx. annulirostris Skuse, Cx. sitiens Wiedemann, and Cx. palpalis Taylor appear to be the most abundant and widespread throughout northern Australia and Papua New Guinea (PNG). Recent investigations using allozymes have shown this subgroup to contain cryptic species that possess overlapping adult morphology. We report the development of a polymerase chain reaction-restriction fragment-length polymorphism (PCR-RFLP) procedure that reliably separates these three species. This procedure utilizes the sequence variation in the ribosomal DNA ITS1 and demonstrates species-specific PCR-RFLP profiles from both colony and field collected material. Assessment of the consistency of this procedure was undertaken on mosquitoes sampled from a wide geographic area including Australia, PNG, and the Solomon Islands. Overlapping adult morphology was observed for Cx. annulirostris and Cx. palpalis in both northern Queensland and PNG and for all three species at one site in northwest Queensland. PMID- 11931040 TI - Cloning and sequence analysis of a cDNA encoding Pso o II, a mite group II allergen of the sheep scab mite (Acari: Psoroptidae). AB - Psoroptes ovis (Hering), the sheep scab mite, is responsible for psoroptic scabies of cattle and sheep. Reverse translation of 30 N-terminal amino acids of the major P. ovis allergen, previously chosen as a candidate immunogen and identified as a 16 kDa protein yielded a degenerate sequence used to design oligodeoxynucleotide polymerase chain reaction (PCR) primers. Use of the PCR primers with a P. ovis cDNA library succeeded in amplification of a 90 bp cDNA gene fragment that was cloned, sequenced, and used to select unique sequencing/PCR primers. Primer walking generated overlapping subclones which yielded the 588 nucleotide consensus sequence of the cDNA encoding the 143 amino acid P. ovis allergen precursor. Nucleotide and translated sequences of the cDNA were compared with sequences in GenBank and found to be homologous to mite group II allergens Lep d II (formerly Lep d I) of Lepidoglyphus destructor Schrank, DerfII of Dermatophagoides farinae Hughes, Der p II of Derrmatophagoides pteronyssinus (Trouessart), Tyr p II of Tyrophagus putrescentiae (Schrank), Eur m II of Euroglyphus maynei (Cooreman) and Gly d II of Glycophagus domesticus (De Geer). The mature P. ovis allergen is composed of 126 amino acids with a calculated molecular mass of 13,468 Da, three disulfide bonds, and pI of 6.06 with one potential o-glycosylation site at Thr116. We designate the P. ovis 16 kDa protein as Pso o II in conformity with nomenclature for mite group II allergens. PMID- 11931041 TI - Larval fly activity on sunlit versus shaded raccoon carrion in southwestern West Virginia with special reference to the black blowfly (Diptera: Calliphoridae). AB - Larvae of Phormia regina (Meigen), Phaenicia sp., and Sarcophaga sp. were identified from raccoon carcasses placed in sunlit and shaded areas at a southwestern West Virginia site in May of 2000. Samples of larvae were taken from each carcass at 3-h intervals over a 153-h experimental period. Phormia regina was clearly the dominant species with large numbers of third instars observed at every 3-h collection period from 81 to 153 h on both carcasses. Mean lengths of third-instar P. regina larvae collected from the sunlit carcass were significantly greater than mean lengths of larvae collected from the shaded carcass. Third-instar Phaenicia sp. also appeared at 81 h on both carcasses, but relatively few (< or = 4) individuals were present in each 3-h collection sample from 81 through 126 h. Larvae of this species were not present in samples from either carcass in those 3-h intervals from 129 to 147 h. Sarcophaga sp. larvae were also collected, but only in samples taken from the sunlit carcass at 81 and 93 h. Ambient temperatures were recorded throughout the experimental period, whereas maggot mass temperatures were not recorded until the appearance of large numbers of second instars at 48 h. From 48 to 69 h, maggot mass temperatures were equivalent to ambient temperatures; but after 69 h, maggot mass temperatures were considerably elevated over ambient temperatures. PMID- 11931042 TI - Amblyomma geochelone, a new species of tick (Acari: Ixodidae) from the Madagascan ploughshare tortoise. AB - The male and female of Amblyomma geochelone n. sp. are described and illustrated by both scanning electron micrographs and line drawings. Specimens of this new tick species were recovered from the endangered ploughshare tortoise, Geochelone yniphora (Vaillant), in northwestern Madagascar. This relatively large tick is morphologically most similar to Amblyomma nuttalli Donitz, which occurs in mainland sub-Saharan Africa where it mainly parasitizes other species of tortoises. However, several characters distinguish the new species from A. nuttalli including the scutal ornamentation in both sexes and the characteristic patterns of shallow grooves on the alloscutum of the female of A. geochelone. Because the adult stages of A. geochelone are almost certainly host specific ectoparasites of the ploughsh are tortoise, this new tick species is also probably endangered. PMID- 11931043 TI - Cancer patients who participate in support groups. PMID- 11931044 TI - Rethinking somatization. PMID- 11931045 TI - Somatization as illness behavior. PMID- 11931046 TI - Symptoms are sufficient: refining our concept of somatization. PMID- 11931047 TI - The time has come to speak of many things. PMID- 11931048 TI - Dealing with dualism. PMID- 11931049 TI - Rethinking somatization. PMID- 11931050 TI - Somatization and the paradigm of psychosomatology. PMID- 11931052 TI - Does the placebo exist? PMID- 11931051 TI - "Secrets" as suppressed or repressed threatening information in the somatizer's autonomic nervous system: problems with implementing the somatization model not with the model per se. PMID- 11931053 TI - Much ado about nothing. PMID- 11931054 TI - A challenge to core beliefs. PMID- 11931055 TI - The powerful placebo: doubting the doubters. PMID- 11931056 TI - Apples, oranges, and placebos: heterogeneity in a meta-analysis of placebo effects. PMID- 11931057 TI - The placebo efficacy study: problems with the definition of the placebo and the mechanisms of placebo efficacy. PMID- 11931058 TI - Differences in electromyographic activity in the multifidus muscle and the iliocostalis lumborum between healthy subjects and patients with sub-acute and chronic low back pain. AB - The present study was carried out to examine possible mechanisms of back muscle dysfunction by assessing a stabilising and a torque-producing back muscle, the multifidus (MF) and the iliocostalis lumborum pars thoracis (ICLT), respectively, in order to identify whether back pain patients showed altered recruitment patterns during different types of exercise. In a group of healthy subjects (n=77) and patients with sub-acute (n=24) and chronic (51) low back pain, the normalised electromyographic (EMG) activity of the MF and the ICLT (as a percentage of maximal voluntary contraction) were analysed during coordination, stabilisation and strength exercises. The results showed that, in comparison with the healthy subjects, the chronic low back pain patients displayed significantly lower (P=0.013) EMG activity of the MF during the coordination exercises, indicating that, over the long term, back pain patients have a reduced capacity to voluntarily recruit the MF in order to obtain a neutral lordosis. In contrast, during the stabilisation exercises, no significant differences between patients and controls were found for the normalised EMG activity of the two muscles. These findings indicated that, during low-load exercises, no insufficiencies in back muscle recruitment were evident in either subacute or chronic back pain patients. During the strength exercises, the normalised activity of both back muscles was significantly lower in chronic low back pain patients (P=0.017 and 0.003 for the MF and ICLT, respectively) than in healthy controls. Pain, pain avoidance and deconditioning may have contributed to these lower levels of EMG activity during intensive back muscle contraction. The possible dysfunction of the MF during coordination exercises and the altered activity of both muscles during strength exercises may be of importance in symptom generation, recurrence or maintenance of low back pain. PMID- 11931059 TI - Thoracolumbar spinal fractures: segmental range of motion after dorsal spondylodesis in 82 patients: a prospective study. AB - In order to study the effect of dorsal spondylodesis on intervertebral movement in patients treated for thoracolumbar fractures, we measured the sagittal range of motion (ROM) in the segments above and below the fractured vertebral body 2 years after operation. Between 1991 and 1996, 82 consecutive patients with a fracture of the thoracolumbar spine (T12, L1, L2 and L3) were treated operatively with open reduction and stabilisation using an internal fixator, combined with transpedicular cancellous bone graft and dorsal spondylodesis. Eighteen T12, 42 L1, 17 L2 and 5 L3 fractures were included. The range of motion of two segments above and two segments below fracture level was measured. This was done on plain flexion and extension radiographs. The data were compared to normal values and to the zero distribution with the Kolmogorov-Smimov test. At all fracture levels the ROM of the segment adjacent to the disturbed endplate of the fractured body was zero (K-S test). All other evaluated segments showed significant loss of ROM (P<0.05) compared to normal values, except segment L1-L2 in L3 fractures (P=0.058). Dorsal spondylodesis at the level of the disturbed endplate in thoracolumbar spinal fractures leads to immobility in this segment, measured on flexion-extension radiographs 2 years after primary operative treatment. More than 50% loss of motion in the two adjacent levels is equivalent to complete loss of ROM in a second segment. PMID- 11931060 TI - Microendoscopic lumbar discectomy versus open surgery: an intraoperative EMG study. AB - This study investigated electromyographic (EMG) activity as a marker of nerve root irritation during two different surgical procedures for lumbar disc herniation. Mechanically elicited EMG activity was recorded during the dynamic stages of surgery in muscle groups innervated by lumbar nerve roots. Confirmation of surgical activity was correlated with the activity of the electromyogram. Fifteen patients with lumbar disc herniations were treated via an endoscopic medial approach, and 15 patients via the open microscopic surgical technique. Results indicated that the endoscopic technique was superior to the open surgical technique and produced less irritation of the nerve root. Significantly less mechanically elicited activity was recorded during both the approach and the root mobilization. The study showed that microendoscopic discectomy allows a smaller incision and less tissue trauma with comparable visualization of the nerve structures than does open surgery. PMID- 11931061 TI - Electromyography of superficial cervical muscles with exertion in the sagittal, coronal and oblique planes. AB - The purpose of the current study was twofold: (1) to determine the isometric force and electromyographic (EMG) relationship of the sternocleidomastoid, splenii and trapezii muscles bilaterally in graded and maximal exertions in the sagittal, coronal and oblique planes. and (2) to develop regression equations to predict force based on the EMG scores. A newly designed and validated cervical isometric strength testing device was used to measure the cervical muscle isometric strength and force/EMG relationship in cervical flexion, extension, bilateral lateral flexion, bilateral anterolateral flexion, and bilateral posterolateral extension, all beginning with an upright seated neutral posture. A group of 40 healthy subjects were asked to exert their cervical motions in the directions of interest, while the force output and EMG from the sternocleidomastoids, splenii, and trapezii were sampled bilaterally at 1 kHz. ANOVA, correlation, and regression analyses were carried out. The force and EMG scores were significantly different between the directions of effort (P<0.01). All regressions were significant (P<0.01). All subjects registered the highest forces in pure extension and the lowest in pure flexion, showing a gradual decrease from the posterior to anterior direction. There was a modest correlation between EMG of the investigated muscles and force (r=0.15-0.76, P<0.01). EMG output was, for example, approximately 66% higher in flexion than in extension (while force output was roughly 30% less in flexion than extension) - thus relatively more muscle activity was required in flexion than extension to generate a given force. The intermediate positions (i.e. anterolateral flexion) revealed force/EMG ratio scores that were intermediate in relation to the force/EMG ratios for pure flexion and pure extension. The cervical muscle strength and cervical muscle EMG are therefore dependent on the direction of effort. PMID- 11931062 TI - Diagnostic validity of somatosensory evoked potentials in subgroups of patients with sciatica. AB - The diagnostic utility of scalp-recorded somatosensory evoked potentials (SEP) in patients with sciatica has generally been regarded as low. The purpose of the present study was to determine the validity of sensory nerve SEP in different subgroups of sciatic patients. A total of 65 consecutive patients with sciatica showing disc pathology and/or facet joint hypertrophy on lumbar computed tomography (CT) and/or myelography were studied. Symptomatic myelographically compressed nerve roots were defined as truly compromised roots. Asymptomatic myelographically normal nerve roots were defined as truly normal roots. Bilateral sensory nerve SEP representing nerve roots L4, L5, and S1 were performed in all patients. Evaluation of SEP included the use of P1 latency inter-root comparison. The false-positive rate of SEP was low. Pathological L4, L5, and S1 SEP therefore strongly indicate true compromise of the corresponding nerve roots. The true positive rate was higher in patients with facet joint hypertrophy with or without additional disc disease than in patients with disc pathology only, and highest if the sciatic sensory symptoms were present during the SEP registration. Diagnostic validity was not influenced by previous episodes of sciatica, the duration of the present episode, or the number of spinal levels with ipsilateral myelographically compressed nerve roots. Pathological SEP strongly indicate sensory radiculopathy in patients with sciatica. Diagnostic efficacy is higher in patients with facet joint hypertrophy than in patients with disc pathology only and highest when the sciatic symptoms are present during registration. PMID- 11931063 TI - Cramp finding: can it be used as a new diagnostic and prognostic factor in lumbar disc surgery? AB - In this prospective study, the validity and the importance of a new finding (cramp finding) in the diagnosis and outcome after lumbar disc surgery were tested. The test is performed with the person in prone position. Against a forceful knee flexion, the examiner holds the leg with one hand and applies a force to overcome the knee flexion. The finding is positive if the examined person feels a disturbing cramp in the leg or thigh. The study was performed between October 1997 and December 1999. Besides the cramp finding, the classical disc herniation examination, including mechanical and neurological findings, magnetic resonance (MR) imaging tests, and laboratory findings were checked pre- and postoperatively. The positive cramp finding in the operated group was 72% (n=133) preoperatively and straight leg raising (SLR) test was positive in all of them. Cramp finding was positive in 70%, 52%, 34%, and 8% of patients postoperatively in the first, 3rd, 12th and 24th months, respectively. The presented finding appears to be as valuable as the SLR, and especially contralateral SLR, tests in lumbar disc surgery. Cramp finding is also important in outcome evaluation. PMID- 11931064 TI - Intradiscal pressure after repeat intradiscal injection of hypertonic saline: an experimental study. AB - Chemonucleolysis with chymopapain is an effective alternative to an operation for the treatment of some patients who have a lumbar intervertebral disc herniation. However, chymopapain is associated with rare but serious complications. Accordingly, alternative substances for chemonucleolysis have been sought. The main beneficial effect of chemonucleolysis derives from the decrease in the intradiscal pressure. We have previously reported that hypertonic saline injected into the intervertebral discs decreased the intradiscal pressure, but only temporarily. The present experimental study investigated changes in the intradiscal pressure after a repeat intradiscal injection of hypertonic saline. The lumbar intervertebral discs of 18 living rabbits were examined: 10% hypertonic saline was injected intradiscally just once in 12 rabbits, and the same dosage was injected again, 4 weeks later, in the same animals. The intradiscal pressure was measured at 1, 4, 8, and 12 weeks after the second injection. The remaining six rabbits were used as controls, without puncture and without injection. The intradiscal pressure of the group with repeat hypertonic saline injection at 4 weeks was significantly lower than that of the control group. The decreased pressure showed a tendency to increase at 8 weeks, and it had recovered at 12 weeks. The results of this study suggest that repeat hypertonic saline injections may be clinically useful. PMID- 11931065 TI - Nucleus pulposus-induced nerve root injury: effects of diclofenac and ketoprofen. AB - MAIN PROBLEM: Nucleus pulposus and/or chronic compression can induce spinal nerve root injury. Inflammation has been proposed as having major importance in the pathophysiologic mechanisms involved in the induction of such injuries. Corticosteroids, potent anti-inflammatory drugs, have been demonstrated to reduce nucleus pulposus-induced spinal nerve root injury. The aim of the present study was to assess the effects of two potent non-steroidal anti-inflammatory drugs (NSAIDs), diclofenac and ketoprofen, in experimental nucleus pulposus-induced spinal nerve root injury in a pig model. METHODS: Eighteen pigs were included in the study. Autologous nucleus pulposus was harvested from a lumbar disc and applied locally around the first sacral nerve root after a partial laminectomy of the first and second sacral vertebrae. Six pigs were treated with daily intramuscular injections of diclofenac, 3 mg/kg body weight, for 7 days. Six other pigs were treated with daily intramuscular injections of ketoprofen, 4 mg/kg body weight, for 7 days. As controls, six pigs received injections with physiologic saline. After 7 days, the pigs were reanesthetized and the nerve conduction velocity over the exposed nerve root area was determined. RESULTS: The nerve conduction velocity was significantly higher in pigs treated with diclofenac than in the saline-treated controls, (57 +/- 6 m/s vs 38 +/- 18 m/s, P<0.05, Student's t-test). The velocity in pigs treated with ketoprofen, 42 +/- 24 m/s, did not differ significantly from that of controls. CONCLUSIONS: This study of two potent NSAIDs indicates that nucleus pulposus-induced nerve root dysfunction may be reduced by diclofenac but not by ketoprofen. The reason for this difference is not known, but it might be related to the fact that ketoprofen and diclofenac belong to different NSAID subgroups and have a different selectivity for the two cyclo-oxygenases COX-1 and COX-2. PMID- 11931066 TI - Proinflammatory cytokines in cerebrospinal fluid and serum in patients with disc herniation and sciatica. AB - Proinflammatory cytokines have been identified in herniated intervertebral discs in humans, and such cytokines have experimentally been demonstrated to be important in the pathophysiological mechanisms of disc herniation. Cerebrospinal fluid (CSF) and serum concentrations of interleukin (IL)-1beta IL-6, IL-8, interferon (IFN)-gamma and tumour necrosis factor (TNF)-alpha were investigated using the enzyme-linked immunosorbent assay (ELISA) technique in 39 patients with lumbar disc herniation and sciatica. Pain duration and pain intensity (visual analogue scale, VAS) were recorded at inclusion, and a clinical examination was performed evaluating neurological findings. The extent of disc herniation (protrusion or extrusion/sequestration) was evaluated perioperatively. Normal concentrations of IL-1beta, IL-6, IFN-gamma and TNF-alpha were present in CSF and serum in almost all patients with lumbar disc herniation. The concentrations of IL-8 in CSF were increased in 12 out of 39 patients, and these increased levels of IL-8 correlated to a short duration of pain and to more pronounced herniation (extrusion or sequestration). No relationship between IL-8 concentrations in CSF and pain intensity, positive neurological findings or a positive straight leg raising (SLR) test was found. The observation of increased concentrations of IL-8 in CSF in patients with a short duration of symptoms supports the concept of the initial involvement of inflammatory mechanisms after a disc herniation. The finding that most of the patients with increased concentrations of IL-8 in CSF had an extrusion or a sequestration may suggest that the increase in IL-8 is related to mechanical nerve root compression, but may also indicate a biochemical effect exerted by the herniated disc on the surrounding tissue. Further studies on the potential role of IL-8 as a biomarker for disc herniation are warranted. PMID- 11931067 TI - Day-case conventional discectomy: a randomised controlled trial. AB - A prospective randomised trial compared the results of conventional lumbar discectomy performed as day cases with those performed on an in-patient basis. The aim of the study was to investigate whether conventional discectomy can be performed safely on a day-case basis. Thirty-one patients were included in the study; 18 were randomised to day-case treatment and 13 to in-patient stay. Day patients mobilised more quickly immediately following surgery and were better able to walk at 2 weeks. The rate of complications was unchanged. It is concluded that conventional discectomy can be undertaken safely as day-case surgery and may indeed speed up the recovery process. PMID- 11931068 TI - Thermal quantitative sensory testing in lumbar disc herniation. AB - The most frequent clinical presentation of sciatica suggests injury of sensor root fibers alone. To evaluate whether thermal quantitative sensory testing (QST) is applicable in the study of sensory dysfunction in lumbosacral disc herniations, 36 men and 30 women with L4/5 or L5/S1 disc herniations underwent thermal QST of the L4, L5, and Si dermatomes. For both levels of disc herniation, there was a significant difference for all temperature modalities, i.e., cold, warmth and heat pain, between all dermatomes as well as between the side of the herniated disc and the corresponding asymptomatic side. However, the proportion of herniated discs classified correctly was only 48% in patients with disc herniations at the L4/5 level, while it was 71% at the L5/S1 level. We conclude that thermal QST measurements reflect and document sensory dysfunction in patients with lumbosacral disc herniation. The method offers a new means both to study the time course of a spontaneous recovery of sensory dysfunction and to evaluate the result of different treatment options. However, thermal QST seems to have the same poor predictive value for identifying the anatomic location of a herniated lumbar disc as conventional electrophysiologic methods. PMID- 11931069 TI - The Adams classification for cadaveric discograms: inter- and intra-observer error in the clinical setting. AB - The Adams classification for discogram morphology is based on a cadaveric study. It provides the basis for several subsequent classifications proposed in the literature. However, little or no attention has been paid to its reproducibility in the clinical setting. The authors assessed the reliability of this classification using three independent observers of differing experience. One hundred and thirty-three discograms belonging to 71 patients with chronic low back pain were reviewed in a randomised and blinded manner. The morphological appearance at each discogram level was assessed and assigned a type according to the Adams classification. The exercise was repeated 3 weeks later. Respective inter- and intra-observer agreements were calculated in the standard fashion using the kappa statistic. Both inter- and intra-observer agreements were excellent (kappa= 0.77-0.85). The Adams grading system for discogram morphology is consistently reproducible amongst observers with differing levels of experience. It can be safely recommended in the clinical setting as a reliable classification. PMID- 11931070 TI - Treatment of thoracolumbar burst fractures by vertebral shortening. AB - Burst fractures may be stable or unstable, so the choice of treatment may be controversial; almost all cases are surgical type. Deciding on the best method and approach is difficult, due to the many possible options and the fact that good results are achieved in only 60-70% of cases. The main problems to be resolved are the residual kyphosis or the recurrence due to loss of reduction. This is a prospective, observational, longitudinal and descriptive study of six patients two men and four women, mean age 46 years - who scored 7 or more points according to the load distribution classification, and were treated with vertebral shortening by a posterior approach and transpedicular fixation with INO plates. Follow-up was for a period of 2 years, and included evaluating pre- and postoperative stability and kyphosis among other data. The results showed a reduction in the mean fracture angle from 17 degrees preoperatively to 1 degrees postoperatively. Full stability was achieved in five patients, and incomplete stability in one patient, who recovered with the use of a corset. There was evidence of arthrodesis in all six patients within 9 months. The use of an anterior approach to treat burst fractures is well recognized; however, treatment with vertebral shortening using a posterior approach has the advantages of less bleeding, shorter surgical time and less residual kyphosis, as a result of putting together two flat surfaces of healthy bone. The residual kyphosis in the present series, after the 2-year follow up, was less than 1 degrees , which is lower than the 5 degrees - 10 degrees reported in the literature. PMID- 11931072 TI - Spine stabilization by psoas muscle during walking and running. PMID- 11931071 TI - Sagittal morphology and equilibrium of pelvis and spine. AB - A prospective analysis of the sagittal profile of 100 healthy young adult volunteers was carried out in order to evaluate the relationship between the shape of the pelvis and lumbar lordosis and to create a databank of the morphologic and positional parameters of the pelvis and spine in a normal healthy population. Inclusion criteria were as follows: no previous spinal surgery, no low back pain, no lower limb length inequality, no scoliotic deviation. For each subject, a 30 x 90-cm sagittal radiograph including spine, pelvis and proximal femurs in standing position on a force plate was performed. The global axis of gravity was determined with the force plate. Each radiograph was digitized using dedicated software. The spinal parameters registered were values for thoracic kyphosis and lumbar lordosis. The pelvic angles measured were: pelvic incidence, sacral slope and pelvic tilt. The global axis of gravity was on average 9 mm anterior of the center of the femoral heads. The anatomic parameter of pelvic incidence angle varied from 33 degrees to 85 degrees (mean: 51.7 degrees, SD: 11 degrees). The average lumbar lordosis was 46.5 degrees. The average thoracic kyphosis was 47 degrees. We found a statistical correlation between incidence angle and lumbar lordosis (r=0.69, P<0.001) and between sacral slope angle and lumbar lordosis (r=0.75, P<0.001). Spine and pelvis balance around the hip axis in order to position the gravity line over the femoral heads. We propose a scheme of sagittal balance of the standing human body. PMID- 11931074 TI - Laser-grid trabeculectomy in rabbits: an experimental in-vivo study. AB - BACKGROUND: Laser-grid trabeculectomy (TE) is a modification of micro-TE designed to enhance the intra- and postoperative safety of filtering surgery by creating several small oval fistulas (max. diameter <200 microm) beneath a scleral flap without peripheral iridectomy. METHODS: After dissecting the conjunctiva and a conventional scleral flap, 10 small perforating fistulas were created in five pigmented rabbits in the region of the gray-white border using an erbium:YAG laser with a beveled side-firing 200 microm endoprobe; conventional TE with iridectomy was performed in another five animals. Anterior chamber inspection and IOP measurements of both eyes were conducted on days 1, 4, and 14 after operation. After the last inspection eyes were saved for morphological analysis. RESULTS: Both procedures (laser-grid and conventional TE) led to a reduction of intraocular pressure in the treated eye compared to the control eye. Differences between the outcomes for the two procedures were not statistically significant. Morphology of the eyes treated by laser-grid TE revealed dense scarring of perforations around the center of the ligamenta pectinata and loose tissue refilling of the more peripheral openings. CONCLUSIONS: The functional outcome of laser-grid TE in rabbits was similar to that of conventional TE in the short term. Histological examination of microperforations in the area of the trabecular meshwork after 2 weeks indicated that tissue repair was less pronounced than in the more anterior perforations. This morphologic finding may be of importance for non-penetrating glaucoma surgery. PMID- 11931073 TI - Indocyanine green angiography in patients with human T cell-lymphotropic virus type 1 uveitis. AB - PURPOSE: To determine the indocyanine green (ICG) angiographic features and to evaluate the choroidal involvement of human T-cell lymphotropic virus type 1 (HTLV-1)-associated uveitis. METHODS: We performed ICG angiography using scanning laser ophthalmoscopy in 54 eyes of 27 patients (8 men and 19 women) diagnosed with HTLV-1 uveitis. The patient's mean age was 51.5 years with a range of 24-65 years. RESULTS: The early phase of ICG angiography revealed ICG leakage from the choroidal vessels in the posterior pole, hyperfluorescent spots that which were not detected with fluorescein angiography, and small hypofluorescent lesions in the macula which most likely corresponded to microcirculatory disturbances in the choriocapillaris. CONCLUSIONS: We suggest that the ICG angiographic findings reflect choroidal lesions such as infiltration with leukocytes and edema. ICG angiography may provide useful information on choroidopathy in HTLV-1 uveitis. PMID- 11931075 TI - Outflow facilities through Descemet's membrane in rabbits. AB - BACKGROUND: The outflow pathway in viscocanalostomy, a new procedure in glaucoma surgery, is unclear; however, outflow through Descemet's membrane has been postulated. This study evaluates outflow rates through Descemet's membrane at different IOP levels in rabbits. METHODS: 51 Descemet's membranes without endothelium from enucleated rabbit eyes were installed in a double-ring system, the Minuth sheet. Different intraocular pressure levels (20, 25, 30, 40, 50 mmHg) were applied to one side of the system. The system was filled with balanced salt solution. The total amount of fluid percolating through Descemet's membrane was measured after 12 h. Based on this, flow rates were calculated. The area of Descemet's membrane was 6.9 mm2. RESULTS: At the pressure of 20 mmHg the flow rate was less than 0.003 microl/min. At pressures above 30 mmHg flow rates ranged from 0.04 microl/min to 0.15 microl/min with a mean of 0.09 microl/min. To achieve pressure control at high pressures, an area of at least 150 mm2 of Descemet's membrane would be needed. CONCLUSION: Descemet's membrane provides good outflow resistance in rabbit eyes. Based on our results for pressure control by outflow through Descemet's membrane only, at least the whole corneal area is needed. If the same is true in humans, additional outflow sources are necessary in cases of viscocanalostomy. PMID- 11931076 TI - Minimizing side effects of ballistic gene transfer into the murine corneal epithelium. AB - BACKGROUND: The beneficial effect of modulating an allospecific immune response by ballistic IL-4 and CTLA4 gene transfer to deliver minimalistic immunologically defined gene expression (MIDGE) vectors into the corneal epithelium was demonstrated in corneal transplantation. However, side effects reduced graft survival in control animals after ballistic transfer without DNA. METHODS: An adapter was constructed for the gene gun apparatus to enlarge and keep constant the distance between the gun and the cornea. Mice were treated by ballistic transfer of luciferase- or IL-10 -encoding MIDGE vectors using gold particles different in quantity, size and size uniformity. Levels of protein expression were determined. Treated corneas were observed under the scanning electron microscope and immunohistologically. Three groups of Balb/c (H-2d) mice received a C3H (H-2 k) corneal graft and two of them had gold particles delivered into the corneal epithelium by gene gun. RESULTS: Using the gene gun and the distance piece, scanning electron microscopy did not reveal morphological differences of the corneal surface compared with untreated corneas on day 2 and 5. Sagittal histological sections of the central cornea did not show an invasion of macrophages 24 h after treatment. The expression of luciferase and IL-10 was not reduced when a smaller amount of gold (0.1 mg instead of 0.5 mg) was employed. Ballistic gold treatment did not reduce graft survival. CONCLUSION: Ballistic gene transfer into the corneal epithelium allows high cytokine expression in the cornea without measurable side effects if an apparatus is used that is adapted for this specific purpose. PMID- 11931077 TI - Pharmacological effects of latanoprost, prostaglandin E2, and F2alpha on isolated rabbit ciliary artery. AB - BACKGROUND: Latanoprost is a prostaglandin (PG)F2alpha analogue widely recognized in the treatment of glaucoma. To investigate the action of this drug on the ocular circulation, we have studied its effects on isolated rabbit ciliary artery. The data obtained on this drug are compared with the data from PGE2 and PGF2alpha. METHODS: Under the microscope, ciliary artery specimens were prepared from rabbit eyes and mounted in a myograph system. The effects of latanoprost, PGE2, and PGF2alpha on the isolated rabbit ciliary artery were investigated in vitro using isometric tension recording methods. RESULTS: Exogenously applied PGF2alpha, but not latanoprost, evoked contraction in the rabbit ciliary artery. After precontraction with excess-[K]0 solution, latanoprost evoked relaxation dose-dependently. Latanoprost at a concentration of 100 microM induced maximum relaxation, which was not blocked by 10 microM L-nitro-L-arginine methylester (L NAME), 1 microM 8-37 calcitonin gene-related peptide (CGRP) or 10 microM indomethacin. Moreover, latanoprost induced relaxation even in preparations without endothelium. The maximum relaxation obtained with PGE2 was somewhat less than 50% of that with latanoprost. CONCLUSIONS: These results indicate that latanoprost and PGE2 relaxed rabbit ciliary artery to different degrees. The relaxation provoked by latanoprost was not dependent on endothelium and was not caused by intrinsic PG, CGRP or nitric oxide. The mechanism of this relaxation is not yet clear. PMID- 11931078 TI - Region-specific pathophysiological alterations occurring in calf lenses in vitro during hyperglycemia. AB - BACKGROUND: The early onset of cataract during diabetes may come about via a variety of pathogenic pathways, but an uncertainty about the significance of each of them exists. METHODS: Calf lenses cultured in a high glucose medium were investigated for regional variations in sorbitol accumulation, changes in lactate dehydrogenase activity, and formation of carbonyl groups in proteins. The results obtained were used to evaluate the contributions of various pathways to the alterations in the lens during hyperglycemia and to relate these findings to morphologically diverse lens substructures. RESULTS: The highest sorbitol accumulation was found in both the anterior and posterior cortex of lenses incubated in hyperglycemic medium. Lactate dehydrogenase activity was strongly affected by high sugar concentration, but the alterations in the equatorial part of lenses were more moderate relative to other substructures. After incubation with glucose, the concentration of Amadori products did not increase significantly compared to non-incubated and incubated controls. Nuclear proteins exhibited the highest level of oxidation. CONCLUSION: The process of sorbitol accumulation is more evident than glycation in the initial stage of hyperglycemia. Lens cortex is affected faster by elevated glucose, while the nucleus is more susceptible to prolonged effects of oxidation, glycation, and glycoxidation. PMID- 11931079 TI - Severe emulsification after combined use of a partially fluorinated alkane and silicone oil. AB - PURPOSE: To present a case with accidental incomplete removal of a partially fluorinated alkane during a direct exchange with silicone oil and a subsequent severe emulsification of the silicone oil. CASE REPORT: During a macular rotation procedure an incomplete exchange of perfluorohexylhexane (F6H6) with silicone oil occurred. Initial postoperative findings showed phase separation and exact visualization of the fundus. After a few days severe opacification of the mixture occurred, markedly reducing fundus visibility. The substances were removed immediately and analyzed by gas chromatography. CONCLUSION: The direct exchange between silicone oil and partially fluorinated alkane should be avoided. Their combined use as a simultaneous endotamponade can cause opaque separation. PMID- 11931080 TI - Suppression of experimental immune-mediated blepharoconjunctivitis in Brown Norway rats by topical application of FK506. AB - BACKGROUND: Experimental immune-mediated blepharoconjunctivitis (EC) in Brown Norway (BN) rats, which is inducible by transfer of antigen-specific T cells, is a model for human allergic conjunctivitis. We investigated the possible inhibition of EC in BN rats by topical application of FK506, which is an immunosuppressive agent that mainly targets T cells. METHODS: To induce EC by active immunization, ovalbumin (OVA) adsorbed to alum was injected into the hind footpads of BN rats. Three weeks after the initial immunization, rats were challenged with OVA by eye drops. Twenty-four hours later, lids including conjunctivas, lymph nodes (LNs), and sera were harvested for histology or reverse transcriptase PCR, proliferation assays, and measurement of IgE titer, respectively. For passive immunization, rats were intravenously injected with 10 million of in vitro-stimulated OVA-primed LN cells. Four days after the transfer, rats were challenged with OVA and evaluated as above. The rats were divided into two groups. One group received topical FK506 treatment three times per day from 15 to 21 days after active immunization or from 1 to 4 days after transfer. The other group was treated with vehicle as above. RESULTS: FK506 treatment suppressed infiltration of both lymphocytes and eosinophils in the conjunctiva either by active or passive immunization (P<0.002). No differences were noted in antigen-specific cellular and humoral immune responses. Concerning cytokine expression in the conjunctiva, a prominent difference was noted only with IL-4, which was more abundantly detected in the vehicle-treated group. CONCLUSION: Topical FK506 treatment suppressed EC in BN rats, possibly by inhibition of IL-4 in the conjunctiva. PMID- 11931081 TI - Neovascularization in the anterior segment of the rabbit eye by experimental anterior ischemia. AB - PURPOSE: To investigate the effects of anterior ischemia accompanied by neither retinal nor choroidal ischemia on the anterior segment of the eye. METHODS: Both long posterior ciliary arteries in the right eye of 14 rabbits were directly cauterized with an electric coagulator. The eyes were enucleated 1, 2, 4, 7, 9 or 14 days after cauterization, then fixed with 4% paraformaldehyde. Semi-thin sections were studied by light microscopy. Several sections were stained with Griffonia simplicifolia lectin, which bound specifically to mammalian vascular endothelium. Other specimens were examined immunohistochemically for vascular endothelial growth factor (VEGF) protein. The tissue specimens of the first postoperative day were studied for expression of VEGF mRNA by in situ hybridization. RESULTS: Atrophy of the iris and ciliary body was seen after the second postoperative day. Corneal neovascularization appeared after 7 days. Neovascularization on the anterior surface of the iris and in the trabecular meshwork was detected after the ninth postoperative day. The proliferative tissues with newly formed vessels obstructed the iridocorneal angle 14 days after the treatment. There was no histological change in either the retina or choroid. Immunohistochemically, VEGF protein was detected in the epithelial and vascular cells of the iris on the first and fourth postoperative day. Expression of VEGF mRNA was detected in the epithelial cells of the ciliary body on the day following the treatment. CONCLUSIONS: Anterior segment ischemia, when unaccompanied by retinal ischemia, causes neovascularization in the cornea, iris and trabecular tissue. PMID- 11931082 TI - Th1/Th2-balance in anterior chamber-associated immune deviation by alloantigen. AB - BACKGROUND: Immune deviation induced by an injection of antigens (Ags) in the anterior chamber of the eye has been termed anterior chamber-associated immune deviation (ACAID). Inoculated Ag induces the generation of T cells that down regulate delayed-type hypersensitivity (DTH). The induction mechanism may well involve various cytokines. METHODS: BALB/c mice were inoculated with C3H/He splenocytes. After a week, subcutaneous immunization was performed in mice with (ACAID group) or without (positive control group) intracameral inoculation of splenocytes. DTH responses were determined by ear-swelling assay a week after subcutaneous immunization. To ascertain which cytokines suppress or promote ACAID induction, the gene transcription levels of various cytokines were evaluated by ribonuclease protection assay in alloantigen-pulsed splenocytes. Enzyme-linked immunosorbent assay was performed to quantitate cytokine production in the culture supernatants. RESULTS: Alloantigen-specific DTH was suppressed in the ACAID group. Interleukin (IL)-2 and interferon-gamma (IFN-gamma) gene transcription levels in the ACAID-induced group were significantly suppressed, but IL-4, IL-5, IL-6, IL-9, and IL-10 gene transcription levels were not different from those in the positive control group. IFN-gamma and IL-2 production in the ACAID group was significantly suppressed compared with that in the positive control group. Increased expression of IL-4 and IL-10 was not detected in the ACAID group compared with the positive control group. CONCLUSION: The results suggest that Th1 suppression of cytokine secretion in the splenic phase plays a role in ACAID induction and Th2-secreting cytokines do not particularly affect ACAID induction by alloantigen. PMID- 11931083 TI - Anomaloscope matches in patients with diabetes mellitus. AB - BACKGROUND: Functional visual deficits can occur in patients with diabetes mellitus who show no visible morphological alterations in the retina. In this study we examined the colour vision of diabetic patients using metameric matches. Patients with and without retinopathy, as well as those who had been treated with laser photocoagulation, were examined to ascertain how the functional alterations in the diabetic eye alter with disease status. METHODS: Rayleigh (red-green) and Moreland (blue-green) metameric matches were determined in a total of 51 diabetic patients (24 patients with no retinopathy, 12 patients with background retinopathy and 15 patients who had undergone laser therapy). Their results are compared to those of a control group of 25 subjects with normal colour vision. RESULTS: A deficit in blue-green colour discrimination found in patients without retinopathy becomes worse with the appearance of vascular alterations in the retina. There is also a significant shift towards green in the Rayleigh match midpoint and towards blue in the Moreland match midpoint, which is at least in part explicable by alterations in lens opacity. Patients who have undergone laser therapy show, on average, better colour discrimination than those with retinopathy, but there is a large variation in their results. CONCLUSION: The alterations in the colour vision of diabetics indicate that at least the early functional changes are occurring at an inner retinal location. Lens opacity changes also play a large role as the age of the patients increases. PMID- 11931084 TI - Optical coherence tomography in the evaluation of incomplete visual acuity recovery after macula-off retinal detachments. AB - PURPOSE: To determine whether pre and/or postoperative optical coherence tomography (OCT) evaluation of the fovea is useful in explaining delayed or incomplete visual acuity recovery after successful surgery for shallow macula-off retinal detachments. METHODS: Prospective study of 16 patients (10 female, 6 male; mean age 53+/-4 years) with a spontaneous rhegmatogenous retinal detachment with peripheral breaks and a shallow elevation of the fovea. Patients were examined by OCT preoperatively and at 1, 6, and 10-12 months after surgery. At 1 month after surgery, fluorescein angiography was also performed. Mean follow-up was 11.0+/-1.0 months. RESULTS: Preoperative OCT images could be classified into four categories according to the extent of retinal edema and presence of foveal depression. The extent of retinal edema in the detached macula did not correlate with preoperative or postoperative vision. In 11 cases, OCT images at 1 month after surgery showed a foveal detachment with residual subretinal fluid that was not visible clinically or on angiography. Long-term persistence of this fluid could be observed in eight cases at 6 months and in one case at 12 months, and was associated with incomplete recovery of visual acuity. CONCLUSIONS: Preoperative OCT of the detached macula can demonstrate widespread retinal edema, the extent of which does not appear to influence final postoperative vision. Delayed and incomplete visual recovery after successful reattachment of the retina may, however, be due to persistent circumscribed subfoveal fluid accumulation that is visible only on OCT not clinically or on fluorescein angiography. PMID- 11931085 TI - Dark adaptation in age-related macular degeneration: relationship to the fellow eye. AB - BACKGROUND: Abnormalities of dark adaptation have been documented in patients with age-related macular degeneration (AMD), but the relationship with the various forms of this disorder has not been studied systematically. METHODS: Dark adapted retinal sensitivities and kinetics of dark adaptation were studied using a Humphrey visual field analyzer adapted for these purposes in patients over 64 years of age. One eye per patient was studied. Study eyes had a normal visual acuity and macular drusen only. The fellow eye was categorized as follows: group I, pigment epithelial detachments and tears of the retinal pigment epithelium (RPE); group II, choroidal neovascularization; and group III, drusen only. The results of psychophysical tests of the study eyes (group I and II and one eye of group III patients) were compared with one another and with older patients without evidence of AMD (group IV). RESULTS: Retinal sensitivity was found to be most consistently abnormal nearest the fovea. The time course of dark adaptation was prolonged beyond 45 min in 10/11 patients (91%) in group I, 6/10 patients (60%) in group II, and 6/10 (60%) in group III and 1/11 (9%) in group IV. CONCLUSION: In a high proportion of patients with visual loss from AMD in one eye, the fellow eye shows abnormal dark adaptation. These changes appear to be most pronounced in patients with detachments of the RPE in the fellow eye. PMID- 11931086 TI - A mutational hot spot in the mitochondrial ND6 gene in patients with Leber's hereditary optic neuropathy. AB - BACKGROUND: Leber's hereditary optic neuropathy (LHON) is a maternally inherited disorder characterized by rapid bilateral loss of central vision. Most patients harbor one of three mutations in the mitochondrial DNA. In order to identify the genetic cause of the disease in one LHON patient without any of the three primary mutations, we sequenced the mitochondrial genome. METHODS: Ophthalmological examination was performed in the affected person and his unaffected relatives. The complete mitochondrial protein coding region was sequenced in the patient. RESULTS: Clinical examination of the affected 10-year-old Turkish boy showed typical features of LHON. Peripapillary microangiopathy was also seen in relatives of the maternal line. Sequence analysis revealed a point mutation at position 14482 in the mitochondrial ND6 gene that changes a conserved methionine residue to isoleucine. A mutation at this nucleotide position has been previously suggested to be of pathogenic significance and has not been detected in any controls. CONCLUSIONS: We have identified that the mutation at nucleotide position 14482 which is the eight mutation in the ND6 gene that causes LHON, making this gene a hot spot for the disease. All eight identified mutations in the ND6 gene lie within the evolutionarily most conserved region of the ND6 gene in a hydrophobic pocket. This may help in understanding the pathomechanism of LHON. PMID- 11931087 TI - On the different connotations of social psychiatry. PMID- 11931088 TI - Adult attachment style. I: Its relationship to clinical depression. AB - BACKGROUND: Although there are an increasing number of studies showing an association of adult attachment style to depressive disorder, such studies have rarely utilised epidemiological approaches with large community-based series and have relied heavily on brief self-report measurement of both attachment style and symptoms. The result is a wide inconsistency in the type of insecure style shown to relate to disorder. The present study examined adult attachment style in a high-risk community sample of women in relation to clinical depression. It utilised an interview measure of adult attachment which allowed for an assessment of both type of attachment style and the degree of insecurity of attachment. A companion paper examines its relationship with other depressive-vulnerability (Bifulco et al. 2002). METHOD: Two hundred and twenty-two high-risk and 80 comparison women were selected from questionnaire screenings of London GP patient lists and intensively interviewed. A global scale of attachment style based on supportive relationships (with partner and very close others) together with attitudes to support-seeking, derived the four styles paralleling those from self report attachment assessments (Secure, Enmeshed, Fearful, Avoidant). In order to additionally reflect hostility in the scheme, the Avoidant category was subdivided into 'Angry-dismissive' and 'Withdrawn'. The degree to which attitudes and behaviour within such styles were dysfunctional ('non-standard') was also assessed. Attachment style was examined in relation to clinical depression in a 12-month period. For a third of the series this was examined prospectively to new onset of disorder. RESULTS: The presence of any insecure style was significantly related to 12-month depression. However, when controls were made for depressive symptomatology at interview, only the 'non-standard' levels of Enmeshed, Fearful or Angry-dismissive styles related to disorder. Withdrawn-avoidance was not significantly related to disorder. CONCLUSION: The relationship of attachment style to clinical depression is increased by differentiating the degree of insecurity of style and differentiating hostile and non-hostile avoidance. PMID- 11931089 TI - Adult attachment style. II: Its relationship to psychosocial depressive vulnerability. AB - BACKGROUND: A range of studies show adult attachment style is associated with depressive-vulnerability factors such as low self-esteem, poor support and childhood adversity. However, there is wide inconsistency shown in the type of insecure style most highly associated. Few studies have examined attachment style in relation to clinical depression together with a range of such factors in epidemiological series. The present study uses an interview measure of adult attachment which differentiates type of attachment style and degree of insecurity of attachment, to see: (a) if it adds to other vulnerability in predicting depression and (b) if there is specificity of style to type of vulnerability. METHOD: Two hundred and twenty-two high-risk and 80 comparison women were selected from questionnaire screenings of London GP patient lists and intensively interviewed. The Attachment Style Interview (ASI) differentiated five styles (Enmeshed, Fearful, Angry-dismissive, Withdrawn and Standard) as well as the degree to which attitudes and behaviour within such styles were dysfunctional ('non-standard'). Attachment style was examined in relation to low self-esteem, support and childhood experience of neglect or abuse, and all of these examined in relation to clinical depression in a 12-month period. RESULTS: The presence of any 'non-standard' style was significantly related to poor support, low self esteem and childhood adversity. Some specificity of type of style and type of vulnerability was observed. Logistic regression showed that non-standard Enmeshed, Fearful and Angry-dismissive styles, poor support and childhood neglect/abuse provided the best model for clinical depression. CONCLUSION: Non standard attachment in the form of markedly Enmeshed, Fearful or Angry-dismissive styles was shown to be associated with other depressive-vulnerability factors involving close relationships, self-esteem and childhood adversity and added to these in modelling depression. PMID- 11931090 TI - Jailhouse Blues revisited. AB - BACKGROUND: Over the past decades a number of studies have appeared on suicide among persons in jail and prison remand settings, and sentenced prisoners. In these papers contradictory views concerning the relation between suicide risk and length of custody were published. METHODS: This article reports on a study that investigated all suicides (n=206) in all 29 Austrian jails and prisons between 1975 and 1996. The authors demonstrate the necessity to calculate the hazard that reflects the suicide risk in relation to time. RESULTS: The results suggest three different periods of high suicide risk: immediately after admission and 2 months thereafter for prisoners on remand; for long-term prisoners the suicide risk correlates with the length of the sentence and slightly increases with the time of custody. CONCLUSIONS: With this knowledge, suicide prevention programmes in prisons and jails could be made more effective and economic. PMID- 11931091 TI - Differential metabolization of the impact of life events on subjects hospitalized for depressive and anxiety disorders: case-control study. AB - BACKGROUND: The study of the relationship between life events and mental illness has progressed considerably over the last 30 years. Practically all mental disorders have been explored using a variety of methods of variable value. The results are in favor of the existence of a temporal link between life events and illness. The part played by life events is the most pronounced in triggering depressive disorders and attempted suicide. Many aspects of this relationship have been explored, but the ability to metabolize the event impact over time has never been studied. This paper reports the results of a longitudinal study. Its objective is to make a comparative study of the ability to metabolize event impacts in depressive in-patients and controls. METHODS: The investigation was designed as a longitudinal, clinical, case-control study constructed on in patients with depressive and anxiety disorders (n=25) and healthy controls from the community (n=28). The absence or presence of mental pathology was assessed using the SCAN, events and event-impacts were reported by subjects from a list of events constructed from the LEDS framework. RESULTS: The results show that the ability to metabolize the event-impact is positively linked to the mental health status of subjects, and that the first self-assessment of the event-impact has a good predictive capacity on the progression of this score a few months later and then 15-18 months later both in in-patients and in controls. PMID- 11931092 TI - Predictors of components of expressed emotion in major caregivers of Turkish patients with schizophrenia. AB - BACKGROUND: Expressed emotion (EE) is a concept reflecting the emotional atmosphere of the home environment. Specific components of EE, namely criticism, hostility and emotional over-involvement, have been found to be important predictors of relapse for schizophrenic patients. The main aim of this study was to examine the predictive power of patient and caregiver characteristics and caregivers' perceptions of frequency, coping, distress/discomfort, control of symptom behaviours by the patient, and attributions on locus of causality for the development of the illness on two components of EE (criticism/hostility and emotional over-involvement) in a sample of major caregivers of Turkish schizophrenic patients. METHODS: Seventy-two caregivers of schizophrenic patients were administered a set of questionnaires tapping socio-demographic and illness related variables, the family questionnaire tapping perceived frequency, distress/discomfort, coping and control of symptom behaviours, causal attributions for illness and, finally, the Expressed Emotion Scale in the hospital setting. RESULTS: The results showed that caregivers' perceptions of coping with specific symptom behaviours decreased criticism/hostility (C/H), whereas perceptions of higher frequency of symptom behaviours increased C/H. For emotional over-involvement (EOI), the number of individuals living in the household, being the mother, father or the spouse, perceptions of coping with symptom behaviours and reported distress/discomfort about symptom behaviours were significant predictors. CONCLUSIONS: Caregivers' perceptions of their ability to cope with symptom behaviours and their reported distress due to these behaviours are important variables related to components of EE and need to be targets in intervention studies. The cultural and clinical implications of the results for the management of schizophrenia and for support for the caregivers are discussed. PMID- 11931094 TI - The use of phenological data to calculate chilling units in Olea europaea L. in relation to the onset of reproduction. AB - The aim of this study was to develop a practical method to evaluate the effective relationship between the amount of winter chilling and the response expressed as the spring reproductive re-starting dates in the olive (Olea europaea L.). Two olive cultivars growing in a special olive orchard in Umbria (central Italy) were studied over a 3-year period (1998-2000): the cultivar Ascolana, typical of central Italy, and the cultivar Giarraffa, typical of southern Italy. The spring reproductive restarts were assessed using data from detailed phenological observations made on 60 trees of each cultivar in an effort to establish the exact date of reproductive bud swelling. The chilling phenomenon was evaluated by using 341 functions derived from a formula developed by researchers at Utah State University to calculate chilling units. The mathematical functions are defined, and show the very close relationship between the amount of winter chilling and the spring reproductive response in the two cultivars in the orchard studied. The results can be used to define the relationship between local climate and plant development, and the mathematical approach can be used to draw maps that can show the suitability of different cultivars on the basis of local climatic conditions. PMID- 11931093 TI - Health care contacts before and after attempted suicide. AB - OBJECTIVE: The prevalence and timing of contact with health care predicting and after a suicide attempt are not well known. This study systematically investigated the pattern of all health care contacts both before and after attempted suicide. METHODS: All consecutive 1,198 suicide attempters treated in hospital emergency rooms in Helsinki during a 12-month period were identified. Data were gathered on all health care contacts 1 year before and after the index attempt. RESULTS: The vast majority of the suicide attempters had contact with health care during the 12 months before and after the index attempt. However, half were without a treatment contact during the final 30 days before the index attempt and one-third in the 30 days following the attempt. Suicide attempters who were not referred to aftercare, did not suffer from a previously recognised major mental disorder, were male, or made non-violent attempts were less likely to be receiving treatment after the attempt. CONCLUSION: Although most suicide attempters have contact with health care within the year before and after the parasuicide, far fewer actually have a treatment contact at the time of the attempt. PMID- 11931095 TI - Statistical downscaling of general-circulation-model- simulated average monthly air temperature to the beginning of flowering of the dandelion (Taraxacum officinale) in Slovenia. AB - Phenological observations are a valuable source of information for investigating the relationship between climate variation and plant development. Potential climate change in the future will shift the occurrence of phenological phases. Information about future climate conditions is needed in order to estimate this shift. General circulation models (GCM) provide the best information about future climate change. They are able to simulate reliably the most important mean features on a large scale, but they fail on a regional scale because of their low spatial resolution. A common approach to bridging the scale gap is statistical downscaling, which was used to relate the beginning of flowering of Taraxacum officinale in Slovenia with the monthly mean near-surface air temperature for January, February and March in Central Europe. Statistical models were developed and tested with NCAR/NCEP Reanalysis predictor data and EARS predict and data for the period 1960-1999. Prior to developing statistical models, empirical orthogonal function (EOF) analysis was employed on the predictor data. Multiple linear regression was used to relate the beginning of flowering with expansion coefficients of the first three EOF for the Janauary, Febrauary and March air temperatures, and a strong correlation was found between them. Developed statistical models were employed on the results of two GCM (HadCM3 and ECHAM4/OPYC3) to estimate the potential shifts in the beginning of flowering for the periods 1990-2019 and 2020-2049 in comparison with the period 1960-1989. The HadCM3 model predicts, on average, 4 days earlier occurrence and ECHAM4/OPYC3 5 days earlier occurrence of flowering in the period 1990-2019. The analogous results for the period 2020-2049 are a 10- and 11-day earlier occurrence. PMID- 11931096 TI - Moderate summer heat stress does not modify immunological parameters of Holstein dairy cows. AB - The study was undertaken during spring and summer months in a territory representative of the Mediterranean climate to assess the effects of season on some immunological parameters of dairy cows. Twenty Holstein cows were used. Eleven of those cows gave birth during spring; the remaining nine cows gave birth in summer. The two groups of cows were homogeneous for parity. Values of air temperatures and relative humidity were recorded both during spring and summer, and were utilized to calculate the temperature humidity index (THI). One week before the expected calving, rectal temperatures and respiratory rates of the cows were recorded (1500 hours), and cell-mediated immunity was assessed by measuring the proliferation of mitogen-stimulated peripheral blood mononuclear cells (PBMC). Within 3 h of calving, one colostrum sample was taken from each cow and analysed to determine content of immunoglobulin (Ig) G1, IgG2, IgM and IgA. At 48 h after birth, passive immunization of the calves was assessed by measuring total serum IgG. During summer, daytime (0900-2000 hours) THI values were above the upper critical value of 72 [75.2, (SD 2.6)] indicating conditions that could represent moderate heat stress. That THI values were able to predict heat stress was confirmed by the values of rectal temperatures and respiratory rates, which were higher (P < 0.05 and P < 0.001 respectively) during summer. Proliferation of PBMC, the colostral concentration of Ig fractions and serum levels of IgG in their respective off-spring did not differ between spring and summer cows. Results indicated that moderate heat stress due to the hot Mediterranean summer does not modify cell-mediated immunity, the protective value of colostrum and passive immunization of the offspring in dairy cows. PMID- 11931097 TI - Association of meteorological and day-of-the-week factors with emergency hospital admissions in Fukuoka, Japan. AB - We carried out a statistical study of the influence of meteorological and day-of the-week factors on the intrinsic emergency patients transported to hospitals by ambulance. Multiple piecewise linear regression analysis was performed on data from 6,081 emergency admissions for 1 year between April 1997 and March 1998 in Fukuoka, Japan. The response variable was the daily number of emergency patients admitted with three types of disease: cerebrovascular, respiratory and digestive diseases. The results showed that the number of emergency patients admitted daily with cerebrovascular disease was significantly associated with temperature on the day of admission and whether the day was Sunday. As it became colder than 12 degrees C, emergency admissions of patients with cerebrovascular disease increased drastically, reaching a plateau at 4 degrees C. On the 3rd and 7th days after the temperature fell below 10 degrees C, the daily admission of patients with respiratory disease significantly increased. We also observed a weak association between emergency admissions of patients suffering from digestive disease and rising barometric pressure on the day of admission. PMID- 11931098 TI - Association between weather conditions and the number of patients at the emergency room in an Argentine hospital. AB - The aim of this paper is to study the relationships between hospital emergencies and weather conditions by analysing summer and winter cases of patients requiring attention at the emergency room of a hospital in the city of Buenos Aires, Argentina. Hospital data have been sorted into seven different diagnostic groups as follows: (1) respiratory, cardiovascular and chest-pain complaints; (2) digestive, genitourinary and abdominal complaints; (3) neurological and psychopathological disorders; (4) infections; (5) contusion and crushing, bone and muscle complaints; (6) skin and allergies and (7) miscellaneous complaints. In general, there is an increase of 16.7% in winter while, for group 2 and group 6, there are more patients in summer, 54% and 75% respectively. In summer, the total number of patients for group 6 shows a significant positive correlation with temperature and dew-point temperature, and a negative correlation with the sea-level pressure for the same day. In winter, the same relationship exists, however its correlation is not as strong. The lags observed between these three variables: maximum dew-point temperature, maximum temperature, minimum air pressure and the peaks in admissions are 1, 2 and 4 days respectively. In winter, increases in temperature and dew point and decreases in pressure are followed by a peak in admissions for group 2. In winter, there are significantly more cases in group 5 on warm, dry days and on warm, wet days in the summer. PMID- 11931099 TI - Changes in mass and energy transfer between the canopy and the atmosphere: model development and testing with a free-air CO2 enrichment (FACE) experiment. AB - The rationale for this study is found in the probable higher temperatures and changes in rainfall patterns that are expected in the future as a result of increasing levels of CO2 in the atmosphere. In particular, higher air temperatures may cause an increase in evapotranspiration demand while a reduction in rainfall could increase the severity and duration of drought in arid and semi arid regions. Representation of the water transfer scheme includes water uptake by roots and the interaction between evapotranspiration and CO2 enrichment. The predicted response of a spring wheat (Triticum aestivum L. cv. Yecora rojo) canopy in terms of energy exchange processes to elevated atmospheric CO2 level was tested against measurements collected at the FACE (Free Air Enrichment Experiment) site in 1994. Simulated and measured canopy conductances were reduced by about 30% under elevated [CO2] under optimum conditions of water supply. Reductions in latent heat fluxes under elevated instead of ambient [CO2] caused reductions in both simulated and measured seasonal water use of 6% under optimum and 2% under suboptimum irrigation. The soil-plant-atmosphere water transfer scheme proposed here offers several advances in the simulation of land surface interactions. First, the stomatal resistance model minimizes assumptions in existing land surface schemes about the effects of interactions among environmental conditions (radiation, temperature, CO2) upon stomatal behavior. These interactions are resolved in the calculation of CO2 in which processes are already well understood. PMID- 11931100 TI - Anionic oligomerization as a route to chain clusters. AB - The combination of two old concepts (the "sigma-donor" capability of M-H bonds and the propensity of unsaturated complexes for addition reactions) has led to the first example of addition oligomerization of an organometallic analogue of olefins: chains containing cis-[Re(CO)4] units joined by Re-H-Re interactions have been obtained through the oligomerization of the unsaturated complex [Re2(micro-H)2(CO)8] promoted by anionic "initiators". The limits of the method and the possible extension to other substrates are discussed. PMID- 11931101 TI - Zinc porphyrin tweezer in host-guest complexation: determination of absolute configurations of primary monoamines by circular dichroism. AB - A nonempirical exciton chirality circular dichroic (CD) method for determining the absolute configurations of primary monoamines with amino group directly linked to the stereogenic center is described. Conventional exciton chirality CD method cannot be applied to these compounds since they lack the two sites for attaching the interacting chromophores. This was solved by covalently linking the monoamine to a trifunctional bidentate carrier moiety 1. Treatment of the carrier/monoamine conjugate with the porphyrin tweezer 4 consisting of two pentanediol-linked zinc porphyrins gives rise to 1:1 host-guest macrocyclic complexes that exhibit exciton-coupled CD spectra. The sign of the CD couplet can then be correlated with the absolute configuration of the monoamine as follows: a clockwise arrangement of the L, M, and S (large, medium, small) groups in the Newman projection of the monoamine with the amino group in the rear gives rise to a positive CD couplet, and vice versa; the assignments of L, M, S groups are based on conformational energies (A values). This method is applicable to cyclic and acyclic aliphatic amines, aromatic amines, amino esters, amides, and cyclic amino alcohols, and can be performed at the several microgram level. PMID- 11931102 TI - Reactivity of ether- and amine-complexed dimers and tetramers of alkyllithiums towards triphenylmethane. AB - Kinetics of Lewis base (LB) complexed primary and secondary sigma-alkyllithiums (RLi) with triphenylmethane (TPMH) are reported. RLis in which one or two LB groups (-OMe, -NMe, -NMeR) are part of the molecule form, in benzene, intramolecularly complexed tetramers, for example, 2(4), or dimers, for example, 4(2). They are used as models for their intermolecular congeners R4Li4 x 4LB and R2Li2 x 4LB (LB = NR'3, OR'2). Nonunity reaction orders in [RLi] are in line with reactions via as yet unidentified 1:1 complexes formed in an equilibrium (K(stat. corr.) approximately = 1) between aggregated RLi and TPMH. In some cases, a tetramer/dimer equilibrium mixture undergoes complexation/reaction. Reaction rates correlate linearly with calculated concentrations of the complexes. Relative rates of complexes range from 1 [prim-R4Li4 x 3LB x TPMH (presumed)] to 4250 [sec-R2Li2 x 3LB x TPMH (presumed)]. A major role in the reactivity enhancement owing to LB-induced conversion of tetramers into dimers is ascribed to increased LB participation in LB-richer dimer transition states. Amine and ether complexes have practically equal reactivities. Lithiation of TPMH by dimeric RCH2Li is retarded by a factor of 24000 if a silyl group is linked to the alpha-carbon. PMID- 11931103 TI - Synthesis, structure, and spectroelectrochemical investigation of novel ternary Co/S(Se)/Sn clusters derived from binary cobalt stannanediyl complexes. AB - The synthesis and structure of heterobimetallic Co/Sn complexes [(eta5-CpR)Co Sn[CH(SiMe3)2]2] (CpR = C5Me5 2; C5EtMe4 3) are described. Insertion reactions of sulfur and selenium into the unbridged heteronuclear Co-Sn bonds of 1, 2, and 3 (R= H5 1, Me5 2, EtMe4 3) have been studied. Depending on the stoichiometry of the chalcogen element used, novel ternary Sn-chalcogen-Co clusters (8, 9, 15, and 16) can be synthesized, and their molecular structures, which represent rare examples of crystallographically characterized cases of ternary transition metal/chalcogen/tin complexes, have been determined. Electrochemistry shows that complexes 8 and 9 are able to support reversibly either the removal or addition of one electron. Insertion of a further (Cp)Co-E (E = chalcogen) fragment significantly affects the electron distribution and causes complexes 9 and 16 to undergo two consecutive one-electron oxidations. The EPR spectra of the respective monocations have been recorded. In all cases, the unpaired electron strongly interacts with the cobalt nucleus(i), thus testifying that the main contribution to the relevant HOMO orbitals comes from the cobalt atom(s). PMID- 11931104 TI - Depsipeptide dendrimers. AB - The convergent synthesis of a new class of chiral dendrimers is described. Owing to their structural resemblance to depsipeptides they are called depsipeptide dendrimers. The ex-chiral pool synthesis starts from (R,R)-, (S,S)-, and meso tartaric acid as branching units and dipeptides or tripeptides consisting of glycine, (L)-alanine, and (L)-leucine as chiral-spacer building blocks. The key intermediates for the convergent assembly of such depsipeptide dendrimers are the peptide-tartaric acid conjugates 13a,b, 19a,b, 25, and 27, which contain either an unprotected C terminus of the peptide chain (13 a,b, 25) or two unprotected hydroxy groups within the tartaric acid termini. Dendra up to the third generation, by using different combinations of stereoisomeric building blocks, were synthesized and completely characterized. Since this construction principle of chiral depsipeptide dendrimers allows for a wide variation of the length, the primary structure of the peptide spacer, and the configuration of both the amino acid and the tartaric acid moieties, access to new combinatorial libraries is conceptually provided. PMID- 11931105 TI - (N-pyrrolyl)B(C6F5)2--a new organometallic Lewis acid for the generation of group 4 metallocene cation complexes. AB - Treatment of the (C6F5)2BF x OEt2 (3) complex with N-pyrrolyl lithium gives bis(pentafluorophenyl)(N-pyrrolyl)borane (2), a strong organometallic Lewis acid, which was characterized by X-ray diffraction (B-N bond length: 1.401(5) A). It exhibits a columnar superstructure in the crystal and contains pi-stacks of pyrrolyl units. Compound 2 readily abstracts alkyl anions from a variety of alkyl Group 4 metallocene-type complexes and leads to the clean formation of the respective metallocene ions or ion pairs. For example, the treatment of Cp3ZrCH3 (9) with 2 transfers a methyl anion to yield the ion pair [Cp3Zr]+[(C4H4N)B(CH3)(C6F5)2]- (12). The X-ray crystal structure analysis of 12 shows a close contact between zirconium and the pyrrolyl-beta-carbon (2.641(2) A). The borane 2 adds to (butadiene)zirconocene (13) to yield the betaine system [Cp2Zr]+[(C4H6)B- (NC4H4)(C6F)2]- (15). Complex 15 contains a distorted eta3 allyl moiety inside the metallacyclic framework and it features an internal Zr+...(pyrrolyl)B- ion pair interaction with a Zr...pyrrolyl-alphacarbon separation of 2.723(3) A (determined by X-ray diffraction). From the dynamic NMR spectra of 15 the bond strength of the internal ion pair interaction was estimated to be deltaGdiss (223 K) approximately = to15 kcalmol(-1). Treatment of dimethylzirconocene (16) with 2 yields the metallocene borate salt [Cp2ZrCH3]+[(C4H4N)B(CH3)(C6F5)2]- (17), which is an active catalyst for the polymerization of ethene. PMID- 11931106 TI - Understanding diastereofacial selection in carbohydrate-based domino cycloadditions: semiempirical and DFT calculations. AB - The sequential cycloaddition of nitroalkenes with methyl vinyl ether was investigated by semiempirical (PM3) and density functional methods (B3LYP/6 31G*). The asymmetric version was also examined with a threoconfigured carbohydrate auxiliary. This produces a larger, more flexible system that complicates the calculation. Most transition structures were then fully optimized at the PM3 level and further refinement was done at ab initio levels. This study represents a model case that enables the rationalization of the high facial selectivity observed in carbohydrate-based nitrone- and nitronate-alkene cycloadditions. The selective endo orientation of the [4+2] pathway results from Coulombic attraction and secondary orbital interactions in the transition state. The stereochemical outcome is largely influenced by a combination of steric shielding from the bulky chiral substituent at C4 and the anomeric effect that places the nitronate C6-alkoxy group in a pseudoaxial arrangement. The resulting conformation favors the subsequent exo approach of methyl vinyl ether to the less hindered re face of the nitronate. It is also remarkable to note that solvation energies stabilize significantly a particular transition structure, thereby explaining the marked stereoselection observed in a polar medium. PMID- 11931107 TI - Enantiospecific synthesis of 1-azafagomine. AB - For the first time the two enantiomeric forms of the glycosidase inhibitor 1 azafagomine have been synthesised starting from D- and L-xylose. D-Xylose was converted to the 2,3,5-tribenzylfuranose, which upon reductive amination with tert-butyl carbazate gave the protected 1-hydrazino-1-deoxypentitol in high yield. N-acetylation, mesylation of the 4-OH, removal of the Boc group, cyclisation and deprotection gave (+)-1-azafagomine ((+)-1). By a similar sequence of reactions, L-xylose was converted to (-)-1-azafagomine ((-)-1). Enzymatic and other routes to optically pure 1-azafagomine were also studied. Compound (-)-1 is a potent competitive glycosidase inhibitor, while (+)-1 has no biological activity. The inhibition of almond beta-glucosidase by (-)-1 was found to be slow owing to a slow binding step of inhibitor to enzyme, with no subsequent conformational rearrangement. The rate constants for binding and release were found to be 3.3 x 10(4)M(-1)s(-1) and 0.011 s(-1), respectively, yielding Ki = 0.33 microM. PMID- 11931108 TI - Enantioselective synthesis of (+)-royleanone from sulfinyl quinones. AB - A convergent enantioselective synthesis of (+)-royleanone (1) is described starting from enantiomerically pure (S)-3-hydroxy-2-isopropyl-5-tert butylsulfinyl-p-benzoquinone, which is readily available from 3-isopropyl-1,2,4 trimethoxybenzene and 1,3,3-trimethyl-2-vinylcyclohexene. The key step is a tandem asymmetric Diels-Alder reaction/pyrolytic sulfoxide elimination process. PMID- 11931109 TI - Lead hydro sodalite [Pb2(OH)(H2O)3]2[Al3Si3O12]2: synthesis and structure determination by combining X-ray rietveld refinement, 1H MAS NMR FTIR and XANES spectroscopy. AB - Ion exchange of the sodium hydro sodalites [Na3(H2O)4]2-[Al3Si3O12]2 [Na4(H3O2)]2[Al3Si3O12]2 and [Na4(OH)]2[Al3Si3O12]2 with aqueous Pb(NO3)2 solutions yielded, whichever reactant sodalite phase was used, the same lead hydro sodalite, [Pb2(OH)-(H2O)3]2[Al3Si3O12]2. Thus, in the case of the non-basic reactant [Na3(H2O)4]2-[Al3Si3O12]2 an overexchange occurs with respect to the number of nonframework cationic charges. Rietveld structure refinement of the lead hydro sodalite based on powder X-ray diffraction data (cubic, a = 9.070 A, room temperature, space group P43n) revealed that the two lead cations within each polyhedral sodalite cage form an orientationally disordered dinuclear [Pb2(micro-OH)(micro-H2O)(H2O)2]3+ complex. Due to additional lead framework oxygen bonds the coordination environment of each metal cation (CN 3+3) is approximately spherical, and clearly the lead 6s electron lone pair is stereochemically inactive. This is also suggested by the absence of a small peak at 13.025 keV, attributed in other Pb2+-O compounds to an electronic 2p-6s transition, in the PbL3 edge XANES spectrum. 1H MAS NMR and FTIR spectra show that the hydrogen atoms of the aqua hydroxo complex (which could not be determined in the Rietveld analysis) are involved in hydrogen bonds of various strengths. PMID- 11931110 TI - 2-rhodaoxetanes: their formation of oxidation of [RhI(ethene)]+ and their reactivity upon protonation. AB - New cationic, pentacoordinate complexes [(TPA)Rh1(ethene)]+, [1a]+, and [(MeTPA)Rh1(ethene)]+, [1b]+, have been prepared (TPA = N,N,N-tri(2 pyridylmethyl)amine, MeTPA = N-[(6-methyl-2-pyridyl)-methyl]-N,N-di(2 pyridylmethyl)amine). Complex [1a]+ is selectively converted by aqueous HCl to [(TPA)RhIII-(ethyl)Cl]+, [2a]+. The same reaction with [1b]+ results in the [(MeTPA)RhIII-(ethyl)Cl]+ isomers [2b]+ and [2c]+. Treatment of [1a]+ and [1b]+ with aqueous H2O2 results in a selective oxygenation to the unsubstituted 2-rho da(III)oxetanes (1-oxa-2-rhoda(III)cyclo-butanes) [(TPA)RhIII(kappa2-C,O-2 oxyethyl)]+, [3a]+, and [(MeTPA)RhIII(kappa2-C,O-2-oxyethyl)]+, [3b]+. The reactivity of 2-rhodaoxetanes [3a]+ and [3b]+ is dominated by the nucleophilic character of their 2-oxyethyl oxygen. Reaction of [3a]+ and [3b]+ with the non coordinating acid HBAr(f)4 results in the dicationic protonated 2-rhodaoxetanes [(TPA)RhIII(kappa2-2-hydroxyethyl)]2+, [4a]2+, and [(MeTPA)RhIII(kappa2-2 hydroxyethyl)]2+, [4b]2+. These eliminate acetaldehyde at room temperature, probably via a coordinatively unsaturated kappa1-2-hydroxyethyl complex. In acetonitrile, complex [4a]2+ is stabilised as [(TPA)-RhIII(kappa1-2 hydroxyethyl)(MeCN)]2+, [5a]2+, whereas the MeTPA analogue [4b]2+ continues to eliminate acetaldehyde. Reaction of [3a]+ with NH4Cl and Mel results in the coordinatively saturated complexes [(TPA)RhIII(kappa1-2-hydroxyethyl)(Cl)]+, [6a]+, and [(TPA)-RhIII(kappa1-2-methoxyethyl)(I)+, [7a]+, respectively. Reaction of [3a]+ with NH4+ in MeCN results in formation of the dicationic metallacyclic amide [(TPA)-RhIII [kappa2-O,C-2-(acetylamino)ethyl]]2+, [9]2+, via the intermediates [4a]2+, [5a]2+ and the metallacyclic iminoester [(TPA)RhIII[kappa2 N,C-2-(acetimidoyloxy)ethyl]]2+, [8]2+. The observed overall conversion of the [Rh(I)(ethene)] complex [1a]+ to the metallacyclic amide [9]2+ via 2-rhodaoxetane [3a]+, provides a new route for the amidation of a [RhI(ethene)] fragment. PMID- 11931111 TI - Hydrogenolysis of benzylic alcohols on rhodium catalysts. AB - Alumina-supported catalysts from various sources and with different rhodium dispersions predominantly yield the hydrogenation product perhydro-1-indanol in the liquid-phase hydrogenation of 1-indanol, while carbonsupported catalysts mainly give the C-O bond scission - hydrogenation product perhydroindane. Addition of organic or inorganic bases to the reaction mixture suppresses C-O bond scission. To distinguish between the two possible pathways for C-O bond scission of direct hydrogenolysis or dehydration followed by hydrogenation, deuteration studies have been carried out with carbon-supported catalysts. Not only 1-indanol but also indane and indene (the two possible mechanistic intermediates in the C-O bond scission routes) were deuterated. Information about the actual pathway has been obtained by determining the degree of deuteration and the positions at which deuterium is incorporated in the resulting perhydroindane product by means of mass spectrometry and 13C NMR spectroscopy. The results prove that C-O bond scission takes place primarily through the direct hydrogenolysis pathway on the carbon-supported catalysts. Direct hydrogenolysis occurs on the carbon support because of the formation of a better leaving group (OH2+) from the benzylic hydroxy group and its subsequent substitution by spillover hydrogen. PMID- 11931112 TI - Formal fusion of a pyrrole ring onto 2-pyridyl and 2-pyrimidyl cations: one-step gas-phase synthesis of indolizine and its derivatives. AB - Two ortho-hetarynium ions, the 2-pyridyl and 2-pyrimidyl cations, react promptly with 1,3-dienes in the gas phase by annulation, formally by fusion, onto the ions of a pyrrole ring. This novel reaction proceeds through an initial polar [4 + 2+] cycloaddition across the C[triple bond]N+ bond, followed by fast ring opening, a [1,4-H] shift, and finally a recyclization that results in a contraction of a six to a five-membered ring and dissociation by the loss of a methyl radical. For the 2-pyridyl cation, this reaction yields ionized indolizines (pyrrolo[1,2 a]pyridines), while for the 2-pyrimidyl cation, it gives ionized pyrrolo[1,2 a]pyrimidines. The annulation reaction, performed in the rf-only collision quadrupole of a pentaquadrupole (QqQqQ) mass spectrometer, occurs readily with both 1,3-butadiene and isoprene, and is thermodynamically and kinetically favored as predicted by ab initio calculations. Ortho-hetarynium ions and 1,3-dienes provide, therefore, the two building blocks for the efficient one-step gas-phase synthesis of ionized bicyclic pyrrolo[1,2-a]pyridine (indolizine) and pyrrolo[1,2 a]pyrimidine, as well as their analogues and derivatives. PMID- 11931113 TI - Asymmetry and magnetism in bis(oximato)-bridged heterobimetallic compounds: a computational approach. AB - A density functional study of exchange coupling was carried out for a series of heterobinuclear oximato-bridged transition metal complexes. Model calculations were used to examine the influence of the electronic configuration of the metal atoms on the coupling constants. This analysis was complemented by a study of the variation of the coupling constant with the most usual structural distortions within this family of compounds. The influence of the nature of the terminal ligands as well as that of the symmetry on the bridge were also investigated. PMID- 11931114 TI - Reactivity of dioxoruthenium(VI) porphyrins toward amines. Synthesis and characterization of bis(arylamine)ruthenium(II), bis(arylamido)- and bis(diphenylamido)ruthenium(IV), and oxo(tert-butylimido)ruthenium(VI) porphyrins. AB - Reactions of dioxoruthenium(VI) porphyrins, [Ru(VI)O2(Por)], with p chloroaniline, trimethylamine, tert-butylamine, p-nitroaniline, and diphenylamine afforded bis(amine)ruthenium(II) porphyrins, [Ru(II)(Por)(L)2] (L-p-ClC6H4NH2, Me3N, Por=TTP, 4-Cl-TPP; L=tBuNH2, Por = TPP, 3,4,5-MeO-TPP, TTP, 4-Cl-TPP, 3,5 Cl-TPP) and bis(amido)ruthenium(IV) porphyrins, [Ru(IV)(Por)(X)2] (X=p-NO2C6H4NH, Por=TTP, 4-Cl-TPP; X = Ph2N, Por = 3,4,5-MeO-TPP, 3,5-Cl-TPP), respectively. Oxidative deprotonation of [Ru(II)(Por)(NH2-p-C6H4Cl)2] in chloroform by air generated bis(arylamido)ruthenium(IV) porphyrins, [RuIV(Por)(NH-p-C6H4Cl)2] (Por=TTP. 4-Cl-TPP). Oxidation of [RuII(Por)-(NH2tBu)2] by bromine in dichloromethane in the presence of tert-butylamine and traces of water produced oxo(imido)ruthenium(VI) porphyrins, [RuVI-O(Por)(NtBu)] (Por=TPP, 3,4,5-MeO-TPP, TTP, 4-Cl-TPP, 3,5-Cl-TPP). These new classes of ruthenium complexes were characterized by 1H NMR, IR, and UV/visible spectroscopy, mass spectrometry, and elemental analysis. The structure of [Ru(IV)(TTP)(NH-p-C6H4Cl)2 . CH2Cl2 was determined by X-ray crystallography. The Ru-N bond length and the Ru-N-C angle of the Ru-NHAr moiety are 1.956(7) A and 135.8(6) degrees, respectively. PMID- 11931115 TI - Unexpected formal [1+3] cycloadditions between azides and alpha-zirconated phosphanes: a route to unprecedented phosphazide and iminophosphorane complexes. AB - Polycyclic zwitterionic complexes that incorporate one or two phosphonium unit(s) as cationic center(s) and zirconocene-ate moiety(ies) as the anionic counterpart(s) can be easily prepared by either [1+3] or [1+3] and [2+3] cycloadditions which involve bi- or tricyclic alpha-zirconated phosphanes 3 or 4 and various azides. Some of these species exhibit unprecedented phosphazide chelation with bonding between the zirconium and a nitrogen atom in the alpha position relative to phosphorus. When heated, the phosphazide complexes lose dinitrogen to form stable polycyclic zwitterionic phosphonium mono- or dinuclear complexes. The solid-state structure of the two zwitterionic complexes 5 and 8 was determined by X-ray crystallography. PMID- 11931116 TI - Platinum-catalysed allylic alkylation: reactivity, enantioselectivity, and regioselectivity. AB - The use of platinum complexes as catalysts for allylic substitution has been studied. A variety of different complexes catalyse the reaction, and several substrates have been tested. In the alkylation of mono(alkyl)-substituted allylic acetates, regioselectivity is highly dependent on ligand choice. By using tricyclohexylphosphine as the ligand, almost complete formation of branched products is observed. The development of a highly enantioselective (ca. 80-90% ee) reaction that makes use of chiral diphenylphosphinooxazoline ligands (abbreviated as (S)-PN) is also described. The enantioselectivity is highly dependent on the ratio of ligand to platinum (when the ratio ligand/Pt is greater than 1:1, the ee drops off dramatically). This is in contrast to palladium and is interpreted in terms of differing coordination chemistry for the two metals ((S) PN is hemilabile when complexed to platinum) and should be of significance to future systems that utilise heterobidentate ligands. The crystal structures of two isoelectronic platinum and palladium complexes [[(S)-PN]MCl2] are also described. PMID- 11931117 TI - A thermal spin transition in [Co(bpy)3][LiCr(ox)3] (ox = C2O4(2-); bpy = 2,2' bipyridine). AB - In the three-dimensional oxalate network structures [M(II)(bpy)3][M(I) M(III)(ox)3] (ox= C2O4(2-); bpy = 2,2'-bipyridine) the negatively charged oxalate backbone provides perfect cavities for tris-bipyridyl complex cations. The size of the cavity can be adjusted by variation of the metal ions of the oxalate backbone. In [Co(bpy)3][NaCr(ox)3], the [Co(bpy)3]2 + complex is in its usual 4T1(t2g5e(g)2) high-spin ground state. Substituting Na+ by Li+ reduces the size of the cavity. The resulting chemical pressure destabilises the high-spin state of [Co(bpy)3]2+ to such an extent that the 2E(t2g6e(g)1) low-spin state becomes the actual ground state. As a result. [Co(bpy)3][LiCr(ox)3] becomes a spin crossover system, as shown by temperature-dependent magnetic susceptibility measurements and single-crystal optical spectroscopy, as well as by an X-ray structure determination at 290 and 10 K. PMID- 11931118 TI - 2-aza-2'-deoxyadenosine: synthesis, base-pairing selectivity, and stacking properties of oligonucleotides. AB - 2-Aza-2'-deoxyadenosine (2, z2Ad) is synthesized via its 1,N6-etheno derivative 7 and enzymatically deaminated to 2-aza-2'-deoxyinosine (3). Compound 2 is converted into the phosphoramidite building block 10b. This is employed in solid phase oligonucleotide synthesis. The 2-azapurine base forms a strong base pair with guanine, but a much weaker one with adenine, thymine, and cytosine. Oligonucleotide duplexes with dangling nucleotide residues, such as 2-aza-2' deoxyadenosine and 7-deaza-2'-deoxyadenosine (4, c7Ad), either on one or both termini, are synthesized, and the thermal stability of the duplexes is correlated with the hydrophobic properties of the dangling nucleotide residues. PMID- 11931119 TI - Visible-light detoxification and charge generation by transition metal chloride modified titania. AB - Amorphous microporous metal oxides of titanium (AMM-Ti) modified with chlorides of PtIV, IrIV, RhIII, AuIII, PdII, CoII, and NiII have been prepared by the sol gel method and characterized by various surface analytical methods. These hybrid AMM-Ti powders are catalysts for the photodegradation of 4-chlorophenol (4-CP) in aqueous solution when illuminated with visible (lambda > or = 400 or 455 nm) or UV (lambda > or = 335 nm) light. The initial rate depends on the dopant level and is highest at 3.0% Pt in the case of PtIV/AMM-Ti. When employed in a photoelectrochemical cell, the activity spectrum of the photocurrent extends downward to about 600 nm, as does the photodegradation of 4-CP. It is suggested that the metal salt acts as a redox-active chromophore, transmitting the photogenerated charges to the amorphous matrix. PMID- 11931120 TI - Surfactant-encapsulated clusters (SECs): (DODA)20(NH4)[H3Mo57V6(NO)6O183(H2O)18], a case study. AB - We present a comprehensive study of the partially reduced polyoxomolybdate [H3 Mo57V6(NO)6O183(H2O)18]21-encapsulated in a shell of dimethyldioctadecylammonium (DODA) surfacmolecules. Treatment of an aqueous solution of (NH4)21[H3Mo57V6 (NO)6O183(H2O)18] . 65H2O (1a) with a trichloromethane solution of the surfactant leads to instant transfer of the encapsulated complex anion into the organic phase. Results from vibrational spectroscopy. analytical ultracentrifugation, small-angle X-ray scattering, transmission electron microscopy, elemental analysis, and Langmuir compression isotherms are consistent with a single polyoxometalate core encapsulated within a shell of 20 DODA molecules. The molar mass of the supramolecular assembly is 20249 gmol(-1) and the diameter is 3.5 nm. A material with the empirical formula (DODA)20(NH4)[H3-Mo57V6NO)6O183(H2O)18] (2) was isolated as a dark violet solid, which readily dissolves in organic solvents. Slow evaporation of solutions of 2 on solid substrates forces the hydrophobic particles to aggregate into a cubic lattice. Annealing these so-formed films at elevated temperature causes de-wetting with terrace formation similar to liquid crystals and block copolymers. Compound 2 forms a stable Langmuir monolayer at the air-water interface; Langmuir-Blodgett multilayers are readily prepared by repeated transfer of monolayers on solid substrates. The films were characterized by optical ellipsometry, Brewster angle microscopy, transmission electron microscopy, and X-ray reflectance. PMID- 11931121 TI - Informed consent in pediatric neurology. AB - Informed consent remains an important but unpopular topic among physicians. It has the ring of a bothersome legal technicality coupled with a time-consuming, procedural requirement that must be obtained principally to satisfy external agencies and avoid malpractice suits. In fact, consent always has been an essential element in the physician-patient relationship, more important as an ethical duty than as a legal requirement, and continues to represent the most important forum of communication between patients and physicians. To explicate informed consent in pediatric neurology, this article begins with an account of consent in adult medical practice with competent patients. Then consent in incompetent patients is discussed and the standards of surrogate decision-making are defined. Next, the complex situation of consent in pediatrics is considered and the concept of assent and dissent of children and how it differs from consent and refusal in adults is introduced. Legal issues of consent are presented and how to resolve conflicts of assent, consent, and refusal between children and parents is discussed. Then, overriding parental refusals of treatment and consent issues in genetic testing are mentioned. Finally, there is a discussion of consent and assent by children for participation in clinical research studies. PMID- 11931122 TI - The minimally conscious state in children. AB - The minimally conscious state (MCS) is a condition of severely altered consciousness in which minimal but definite behavioral evidence of self- or environmental awareness is shown. Diagnostic criteria recently have been proposed for entry into and emergence from the MCS. We present clinical and neuroimaging data on 5 children diagnosed with MCS and discuss the limited information available concerning its epidemiology, etiology, pathology, and prognosis. Issues related to the evaluation and care of children suspected of having MCS are also reviewed as well as current ethical and legal controversies. PMID- 11931123 TI - The relevance of moral theory to pediatric neurology. AB - Although you should never abandon your well-considered moral judgments because of their conflict with a moral theory, you should also usually not hold that your well-considered moral judgments are the only correct moral judgments. Common morality does not provide unique correct answers to every moral question, it only sets limits to the range of morally acceptable answers. In serious discussions of controversial cases or policies, almost all of the answers will be morally acceptable. The recognition that there are sometimes several conflicting morally acceptable answers allows for a friendly and fruitful discussion with no one sacrificing their moral integrity. PMID- 11931124 TI - Ethical issues in neonatal intensive care: perspectives for the neurologist. AB - Intensive medical care for neonates may present medically complex and ethically challenging issues affecting not only patients but also their families and the professionals caring for them. The consulting neurologist working with these infants and their families is frequently a key figure in determining diagnoses and prognoses. This places the neurology specialist at the center of the process by which the treating team provides parents with appropriate information, collaborates with them in making medical and ethical decisions, and supports them through immeasurably difficult times. PMID- 11931125 TI - Reflections on futility. AB - Medical futility concerns itself with decisions regarding treatments that are of minimal potential benefit to the patient. The concept of futility has evolved with the practice of medicine and consensus has been elusive. Presently, most typically, futility is conceptualized quantitatively as overwhelming improbability in the face of possibility, and qualitatively as an outcome that although achievable is overwhelmingly poor from the patient's perspective. Though frequently intermixed, futility and rationing are separate matters, with futility reflecting individual decisions taken at the bedside, whereas rationing represents decisions of publicly determined health policy. Objective, uniformly applicable guidelines regarding futility will likely be difficult to achieve and thus local efforts must be focused on implementing mechanisms to minimize and resolve inevitable disputes between patients, families, and caregivers. PMID- 11931126 TI - Ethics of clinical research in children. AB - The ethical conduct of clinical research in children is challenged by the inherent vulnerability of this population. Children are both incompetent decision makers and have a right to custody (ie, protection), yet they must also be participants in research that will advance the care of pediatric disorders or they risk becoming "therapeutic ophans." Consensus guidelines exist for the conduct of clinical research in children. Aspects of consent, risk-benefit ratios, fair subject selection, and respect for subjects have child-specific concerns. It is only by explicitly recognizing and addressing these child specific concerns can the conduct of pediatric clinical research achieve and maintain a high level of ethical propriety. PMID- 11931127 TI - Ethical dimensions of genetics in pediatric neurology: a look into the future. AB - Health care providers and families with children who participate in genetic research or who need specialized genetic services, including genetic testing, will encounter not only medical but difficult social, ethical, and legal questions surrounding pediatric genetic neurology. Children are often at the center of much of the genetic revolution and their unique needs raise special concerns about the risks and benefits associated with genetic research, particularly the issues of consent, the use of genetic databases, and gene therapy. Moreover, genetic research and testing raise important psychosocial risks. In this article we discuss some of the benefits and consequences of genetic technologies for children in relation to national and international guidelines. In particular, physicians, policy-makers, and families should be knowledgeable about the guidelines and have a good understanding of the psychosocial and ethical issues associated with genetics in pediatric neurology. PMID- 11931128 TI - Teaching ethics to trainees. AB - Medical events contain many sources of uncertainty. Instruction in medical ethics gives trainees vital knowledge that assists them with some of the sources of uncertainty. Instruction to provide this knowledge is not available from the usual science curriculum. From an understanding of the principles of ethics flows an application of those principles to timely ethical issues. Ethics education has become a mandated feature in accredited residency training programs. A flexible curriculum in ethics can be developed for trainees in child neurology. The education program encourages thoughtful caregivers and teaches the methods for resolution of ethical issues and conflicts. Selection of topics is based on faculty and trainee interest and on situations of current relevance in the training program. Ethics education results in trainees who become capable of ethical reasoning and capable of resolving many of the clinical situations where issues arise that require decisions based on ethical principles. PMID- 11931129 TI - Ethical issues in clinical practice: cases and analyses. AB - Given the severity and complexity of the neurologic disorders that can affect infants and children, pediatric neurologists frequently encounter complex and challenging ethical issues. This discussion uses five cases to illustrate common ethical issues and dilemmas that are confronted by pediatric neurologists. These cases involve the provision of artificial nutrition and hydration to patients in vegetative or minimally conscious states, the use of mechanical ventilation for patients with spinal muscular atrophy, controversial therapies for autistic spectrum disorders, the use of medical information obtained over the Internet, and genetic testing in children with suspected Huntington's disease. The ethical concepts discussed in these cases include autonomy, confidentiality, futility, surrogate decision-making, the best-interest principle, and the withholding and withdrawal of treatment. PMID- 11931130 TI - Materials, methods, results and conclusions. PMID- 11931131 TI - Mutans streptococci and lactobacilli in saliva after the application of fissure sealants. AB - Because dental fissures may serve as reservoirs for mutans streptococci, preventive measures should be taken to control microbial concentrations at these sites. This study estimated the influence that sealing permanent first molars would have on the levels of mutans streptococci and lactobacilli in saliva of healthy seven-year-old schoolchildren. Permanent first molars were sealed in 31 children without caries (NC group) and in 32 children with caries (C group). None of the children had caries in their permanent first molars. Conventional methods were used to count mutans streptococci and lactobacilli in saliva before applying the sealant and again at 4 and 12 weeks after application. Baseline counts of lactobacilli and mutans streptococci were significantly higher in the C group. A significant reduction in mutans streptococci was observed in the NC group at 4 and 12 weeks. It was concluded that fissure sealants in permanent first molars can help reduce salivary levels of mutans streptococci in children without caries. PMID- 11931132 TI - One-year clinical performance of a resin-modified glass ionomer and a resin composite restorative material in unprepared Class V restorations. AB - This study evaluated the clinical performance and appearance of a resin-modified glass ionomer and a resin composite over one year. Thirty-seven pairs of restorations of Fuji II LC and Z250/Single Bond were placed in caries-free cervical erosion/abfraction lesions without tooth preparation. Restorations were clinically evaluated at baseline and 6 and 12 months, using modified Ryge/USPHS criteria. No significant difference (p>0.05) was observed in performance of both materials, although retention of the Z250 restorations was below the minimum specified in the ADA Acceptance Program for Dentin and Enamel Adhesives. Little difference in the restorations' appearance was observed. PMID- 11931133 TI - Placement and replacement of restorations in general dental practice in Iceland. AB - Ninety-one Icelandic practicing dentists (51% response rate) provided information related to the reasons for placement and replacement of 8,395 restorations and 741 sealants in 5,997 patients. Information included the patient's gender and age, the clinician's gender and experience in years since graduation, the defined criteria for replacement of restorations, the estimated past use of material in five-year increments and the records of 100 consecutively placed restorations. The materials used include composite (52.7%), amalgam (29.2%), glass ionomer (9.5%), resin-modified glass ionomer (7.1%) and other materials (1.4%). Although material selection was independent of the clinician's gender, female patients received more composite and fewer amalgam restorations than their male counterparts. Reasons for placing restorations comprised replacement of failed restorations (47.2%), primary caries (45.3%) and non-carious defects (7.5%). Secondary caries was the main reason for replacement for all types of restorations. Chi square analysis related to the dependence between the reasons for replacement and clinician's experience showed that more experienced clinicians recorded a lower frequency of secondary caries than less experienced ones (p<0.0001), while the diagnoses of discoloration and fracture of restorations increased with the clinicians' experience (p<0.0001). PMID- 11931135 TI - Marginal adaptation of ceramic inserts after cementation. AB - The advantage of using ceramic inserts is to prevent major drawbacks of composite resins such as polymerization shrinkage, wear and microleakage. This in vitro study evaluated the marginal adaptation of two approximal ceramic insert systems after cementation to the cavities opened with ultrasonic tips. Proximal cavities with margins in enamel were prepared in 20 intact molars using ultrasonic tips (SONICSYS approx tips [microtorpedo size #2 and #3]; Siplus Instrument approximal [U-shaped]). Inserts of similar sizes (n=10) from two systems corresponding to the ultrasonic tips were placed in the cavities (SONICSYS Inlay; SDS-Inlay system), one on the mesial side and the other on the distal side of the same molar. Following cementation and thermocycling (5000 cycles, between 5-55 degrees C), cement thickness was measured at the buccal, lingual walls and pulpal floors of the proximal boxes under light microscope (x150). The mean cement thickness values recorded for SONICSYS inserts #2 (25 microm) was not significantly different (p>0.05) from that of SDS inserts of similar size (26 microm). There was a significant difference (p<0.05) in cement thickness values between SONICSYS #3 inserts (34 microm) and SDS inserts of similar size (23 microm). Comparison of mean values between the ceramic insert systems examined revealed that marginal adaptation was better at the buccal and lingual proximal walls than those at the pulpal floor in the SDS system, however, there was no difference for SONICSYS at both sizes. Ceramic inserts placed in cavities prepared with ultrasonic tips provide clinically acceptable marginal quality. PMID- 11931134 TI - Penetrability of dentinal tubules in adhesive-lined cavity walls. AB - This study investigated the penetrability of dentinal tubules in cavity walls lined with different dentin bonding systems. Occlusal Class I cavities were prepared in 93 premolars. The cavities in the control group had an intact smear layer without a lining, while those in the experimental group were lined with Gluma CPS, Scotchbond Multi-Purpose Plus or One-Step. The penetrability of the dentinal tubules was tested with a dye (basic fuchsin) or bacteria (S faecalis) immediately after adhesive lining and after one-month storage in water at 37 degrees C. Some of the lined samples were sectioned and examined under the SEM. In some samples in the experimental group, the dye penetrated to the pulp and bacteria up to 125 microm into the dentinal tubules immediately after lining. The Kruskal Wallis ANOVA and Tukey test showed the depth of dye and bacterial penetration to be significantly less in teeth with bonding systems than those in the control group (p<0.05). However, there was no statistically significant difference between the control and experimental groups after storage in water (p>0.05). SEM examination showed that the hybrid layer and resin tags were present in the cavity walls immediately after lining but absent after storage in water. Therefore, adhesive linings under the experimental conditions were ineffective in preventing dye or bacterial penetration of the dentinal tubules. PMID- 11931136 TI - The influence of dental alloys on three-body wear of human enamel and dentin in an inlay-like situation. AB - This in vitro study evaluated the effect of metal alloys on three-body wear resistance of enamel and dentin, and vice versa. Three-body wear of human enamel, dentin, a soft gold alloy (BiOcclus Inlay), a CoCr alloy (Remanium 2000), a resin cement (Variolink II) and a zinc oxide phosphate cement (Harvard) was investigated using the ACTA-machine. Sample chambers of eight sample wheels were prepared with pure materials or combinations of human tooth substance, alloys and cement, simulating an inlay-like situation. After 100,000 and 200,000 cycles in a millet suspension with a spring force of 20 N, the amount of abraded material was profilometrically measured and evaluated by 3D surface data analysis. After 200,000 cycles, the materials demonstrated a mean loss of 0.41 microm for CoCr, 51 microm for gold, 57 microm for enamel, 164 microm for dentin, 79 microm for Variolink and 369 microm for Harvard. Using ANOVA and the Games-Howell-test, resin cement, enamel and gold were a subset not shown to differ, as was zinc phosphate cement and dentin. CoCr demonstrated the least wear and differed significantly from all materials. Enamel wear was significantly reduced in mixed chambers with CoCr, and gold after 200,000 cycles compared to enamel in pure chambers. In summary, a soft gold alloy can be recommended for inlays when considering three-body abrasion since the wear rate of the "soft" gold alloy corresponded to that of human enamel. PMID- 11931137 TI - Finishing and polishing of a hybrid composite and a heat-pressed glass ceramic. AB - This study assessed the finishing and polishing of a hybrid composite and a glass ceramic. Ninety Tetric specimens were divided into three groups of 30 specimens and finished with three different finishing procedures. The 30 specimens were subsequently subdivided into six groups of five and polished using the following polishing systems: Sof-Lex disks, the Ceramiste kit, a diamond polisher, Diafix oral, the MPS gel and the Politip system. Seventy-five IPS-Empress specimens were divided into groups of 25 and finished with three different procedures. The 25 specimens were then subdivided into five groups of five and polished with the same systems, except for the Politip technique. The polished surfaces were evaluated quantitatively by laser stylus profilometry with respect to Ra and profile-length-ratio (LR). Qualitative assessment was carried out by SEM. Quantitative results were examined statistically by one- and two-way Anova and LSD test with significance level of 0.05. The lowest roughness on composite specimens was achieved by the MPS gel and Diafix after finishing according to FM 3 and FM 2. With respect to all methods used, there were no significant differences among the five methods with the lowest Ra-values. The ceramic specimens were able to be polished to lower roughness values (p<0.001 for LR). The best results on ceramic surfaces were achieved with the MPS system after finishing according to FM 3 and FM 2. There were no significant differences among the three methods with the lowest Ra-values and the glazed surface. SEM evaluation largely confirmed the quantitative results. Composite specimens exhibited signs of selective resin matrix removal when the Ceramiste system or the Politip system were used. PMID- 11931138 TI - Influence of bonding agent composition on flexural properties of an Ultra-High Molecular Weight Polyethylene Fiber-Reinforced Composite. AB - This study evaluated the influence of seven commercially available bonding agents on the flexural properties of an Ultra High Molecular Weight Polyethylene (UHMWPE) Fiber-Reinforced Composite (FRC). Nine groups (n=10 per group) of flexural strength specimens were prepared from an indirect composite reinforced with UHMWPE fiber and cured according to manufacturers' instructions (Groups A to I). Group I was a fiber-reinforced negative control without any bonding agent resination. A tenth group (Group J) was a positive control group prepared using indirect composite alone. The fiber reinforcement material for Groups A to H was resinated with one of the seven different bonding agents. Group H used the same bonding agent as for Group G specimens. However, the fiber was silanized before bonding agent application for Group G specimens. Specimens were stored wet for 24 hours at 37 degrees C before measuring flexural strength and modulus in three point bend at a crosshead speed of 1 mm/minute. Scanning Electron Microscopy (SEM) was employed to assess the fiber-resin interface of representative samples. The mean (SD) flexural strength of the test groups impregnated by the bonding agents ranged from 169 (37) to 266 (39) MPa. Statistical analysis of the flexural strength data using one-way ANOVA revealed significant (p<0.05) differences between the test groups. There was catastrophic fiber/composite failure in the positive control group that had a mean flexural strength of 75 (8) MPa. Silane pre-treatment of UHMWPE fiber before impregnation with the bonding agent significantly reduced the flexural strength (p<0.05). PMID- 11931139 TI - Amalgam repair: evaluation of bond strength and microleakage. AB - This study evaluated the tensile bond strength of "repaired" amalgams and compared the degree of microleakage. Amalgam (Cavex avalloy) was condensed into plastic tubes (3 mm in diameter, 10 mm in height) to the half-length. After storage in water at 37 degrees C for two days, the remaining parts of tubes were filled with amalgam (A), cavity varnish (CV)+A, Liner Bond 2V (LB2V)+A, 3M Opal Luting Cement (3MOLC)+A, Panavia F(PF)+A, Metabond(MB)+A, Fuji BondLC(FB)+A, HytacOSB(HOSB)+Hytac Aplitip (H), Liner Bond2V+Clearfil AP-X(CAP). The bond strengths for 15 samples of each restoration group were determined. For the microleakage study, MOD cavities of 90 extracted human premolars were used. The distal half of cavities were filled with amalgam. After storage in water at 37 degrees C for two days, the mesial half of the cavities were filled to simulate a clinical repair. The "repair" was placed using the procedures applied in the bond strength study. The teeth were stained with basic fuchsine (0.5%), sectioned and evaluated for dye penetration. In both parts of study, the data were analyzed by ANOVA and Duncan's multiple range tests. Bond strength values (MPa) were: A+PF+A 3.84+/-1.08, A+LB2V+A 3.15+/-0.97, A+LB2V+CAP 3.05+/-0.53, A+MB+A 2.86+/-0.88, A+HOSB+H 2.58+/-0.51, A+3MOLC+A 2.11+/-0.75, A+FB+A 0.68+/-0.59. The repaired A+A and A+CV+A groups were separated before testing. The A+PF+A group showed the highest bond strength (p<0.05). Microleakage in the cervical margins of repaired restorations was lower in the amalgam groups than microleakage in the resin composite and compomer groups. PF, MB, 3MOLC and FB performed better at the amalgam "repair" interface. The A+LB2V+A group showed no microleakage at both the occlusal and gingival test regions. PMID- 11931140 TI - Rx for caries prevention: time line for home care. A software aid for communication of patient instructions for management of dental caries. AB - Treatment of a caries active patient requires the clinician to be knowledgeable in the restorative and risk management issues with which these patients often present. Modifications of risk factors that promote dental caries play an important part in the long-term restorative success in these patients. Patient compliance to recommendations addressing modifiable caries risk factors for maintenance of oral health is important to long-term caries risk management. Effective dentist-patient or hygienist-patient communication is a first step to encouraging a proactive role by the patient. Others have reported verbal communication accompanied by written communication to result in better compliance compared to verbal communication, alone (Tagliacozzo & Ima, 1970; MacDonald, MacDonald & Phoenix, 1977; Baker & others, 1991; Harvey & Plumridge, 1991; Vukmir & others, 1993; Makoul, Arntson & Schofield, 1995). Further evidence shows that medical patients who misunderstand their treatment plans and associated instructions usually exhibit poor compliance to following directions (Cargil, 1992; Counsell, Geddis & Smith, 1993; Vukmir & others, 1993; Makoul & others, 1995; Mayeaux & others, 1996). Computer-generated (printed) instructions were shown to significantly increase compliance to emergency room instructions when compared to verbal or verbal + hand-written instructions (Vukmir & others, 1993). Written instructions for use in management of dental caries have not been widely addressed in the literature (Fuller & Harding, 1991; Benn & others, 1997). This report presents a novel computer-generated, graphic approach to written communication that recognizes the patient's need to be reminded that home care consists of daily cyclic events. The objective of this approach is to promote communication with and comprehension by a wider population range than what might be affected by text-only instructions. The software for this method is pre-loaded with recommendations that can be modified by the dentist or hygienist and can be easily adapted as part of a comprehensive disease management program used in the office. PMID- 11931141 TI - Notification that new names and new combinations have appeared in volume 52, part 1, of the IJSEM. PMID- 11931142 TI - The phagotrophic origin of eukaryotes and phylogenetic classification of Protozoa. AB - Eukaryotes and archaebacteria form the clade neomura and are sisters, as shown decisively by genes fragmented only in archaebacteria and by many sequence trees. This sisterhood refutes all theories that eukaryotes originated by merging an archaebacterium and an alpha-proteobacterium, which also fail to account for numerous features shared specifically by eukaryotes and actinobacteria. I revise the phagotrophy theory of eukaryote origins by arguing that the essentially autogenous origins of most eukaryotic cell properties (phagotrophy, endomembrane system including peroxisomes, cytoskeleton, nucleus, mitosis and sex) partially overlapped and were synergistic with the symbiogenetic origin of mitochondria from an alpha-proteobacterium. These radical innovations occurred in a derivative of the neomuran common ancestor, which itself had evolved immediately prior to the divergence of eukaryotes and archaebacteria by drastic alterations to its eubacterial ancestor, an actinobacterial posibacterium able to make sterols, by replacing murein peptidoglycan by N-linked glycoproteins and a multitude of other shared neomuran novelties. The conversion of the rigid neomuran wall into a flexible surface coat and the associated origin of phagotrophy were instrumental in the evolution of the endomembrane system, cytoskeleton, nuclear organization and division and sexual life-cycles. Cilia evolved not by symbiogenesis but by autogenous specialization of the cytoskeleton. I argue that the ancestral eukaryote was uniciliate with a single centriole (unikont) and a simple centrosomal cone of microtubules, as in the aerobic amoebozoan zooflagellate Phalansterium. I infer the root of the eukaryote tree at the divergence between opisthokonts (animals, Choanozoa, fungi) with a single posterior cilium and all other eukaryotes, designated 'anterokonts' because of the ancestral presence of an anterior cilium. Anterokonts comprise the Amoebozoa, which may be ancestrally unikont, and a vast ancestrally biciliate clade, named 'bikonts'. The apparently conflicting rRNA and protein trees can be reconciled with each other and this ultrastructural interpretation if long-branch distortions, some mechanistically explicable, are allowed for. Bikonts comprise two groups: corticoflagellates, with a younger anterior cilium, no centrosomal cone and ancestrally a semi-rigid cell cortex with a microtubular band on either side of the posterior mature centriole; and Rhizaria [a new infrakingdom comprising Cercozoa (now including Ascetosporea classis nov.), Retaria phylum nov., Heliozoa and Apusozoa phylum nov.], having a centrosomal cone or radiating microtubules and two microtubular roots and a soft surface, frequently with reticulopodia. Corticoflagellates comprise photokaryotes (Plantae and chromalveolates, both ancestrally with cortical alveoli) and Excavata (a new protozoan infrakingdom comprising Loukozoa, Discicristata and Archezoa, ancestrally with three microtubular roots). All basal eukaryotic radiations were of mitochondrial aerobes; hydrogenosomes evolved polyphyletically from mitochondria long afterwards, the persistence of their double envelope long after their genomes disappeared being a striking instance of membrane heredity. I discuss the relationship between the 13 protozoan phyla recognized here and revise higher protozoan classification by updating as subkingdoms Lankester's 1878 division of Protozoa into Corticata (Excavata, Alveolata; with prominent cortical microtubules and ancestrally localized cytostome--the Parabasalia probably secondarily internalized the cytoskeleton) and Gymnomyxa [infrakingdoms Sarcomastigota (Choanozoa, Amoebozoa) and Rhizaria; both ancestrally with a non-cortical cytoskeleton of radiating singlet microtubules and a relatively soft cell surface with diffused feeding]. As the eukaryote root almost certainly lies within Gymnomyxa, probably among the Sarcomastigota, Corticata are derived. Following the single symbiogenetic origin of chloroplasts in a corticoflagellate host with cortical alveoli, this ancestral plant radiated rapidly into glaucophytes, green plants and red algae. Secondary symbiogeneses subsequently transferred plastids laterally into different hosts, making yet more complex cell chimaeras--probably only thrice: from a red alga to the corticoflagellate ancestor of chromalveolates (Chromista plus Alveolata), from green algae to a secondarily uniciliate cercozoan to form chlorarachneans and independently to a biciliate excavate to yield photosynthetic euglenoids. Tertiary symbiogenesis involving eukaryotic algal symbionts replaced peridinin containing plastids in two or three dinoflagellate lineages, but yielded no major novel groups. The origin and well-resolved primary bifurcation of eukaryotes probably occurred in the Cryogenian Period, about 850 million years ago, much more recently than suggested by unwarranted backward extrapolations of molecular 'clocks' or dubious interpretations as 'eukaryotic' of earlier large microbial fossils or still more ancient steranes. The origin of chloroplasts and the symbiogenetic incorporation of a red alga into a corticoflagellate to create chromalveolates may both have occurred in a big bang after the Varangerian snowball Earth melted about 580 million years ago, thereby stimulating the ensuing Cambrian explosion of animals and protists in the form of simultaneous, poorly resolved opisthokont and anterokont radiations. PMID- 11931143 TI - Phylogenetic analysis of Xanthomonas species based upon 16S-23S rDNA intergenic spacer sequences. AB - Phylogenetic relationships of 17 species of Xanthomonas were assessed based on analysis of 16S-23S rDNA intergenic spacer (ITS) sequences; a higher level of resolution was obtained than that revealed by 16S rDNA analysis. ITS sequences varied in size from 492 to 578 nt within the genus and the similarity among sequences ranged from 63 to 99%. Major differences were found for the hyacinthi group, which included Xanthomonas albilineans, Xanthomonas hyacinthi, Xanthomonas sacchari, Xanthomonas theicola and Xanthomonas translucens. A common ITS structure with tRNA(Ala) and tRNA(Ile) embedded within the sequence was found for all ITS sequences of Xanthomonas species and for Stenotrophomonas maltophilia. These tRNAs were highly conserved and divided the ITS sequence into three regions (ITS1, ITS2 and ITS3). ITS1 and ITS2 sequences of Xanthomonas species showed mean similarities of 87.1 and 86.8%, respectively, and differences consisted of substitution and addition/deletion of 1-5 nt. ITS2 showed remarkable variation in sequence length: most species had an ITS2 of 19-20 nt, whereas a long insertion of 51-56 nt was found in Xanthomonas codiaei, X. hyacinthi, Xanthomonas melonis, X. theicola and X. translucens. For ITS3 the most striking alteration was seen in X. hyacinthi, which showed a large deletion of 44 nt. The ITS phylogenetic tree grouped Xanthomonas species into six major clusters. Cluster I included Xanthomonas arboricola, Xanthomonas axonopodis, Xanthomonas bromi, Xanthomonas campestris, X. campestris pv. gardneri, Xanthomonas cassavae, X. codiaei, Xanthomonas cucurbitae, Xanthomonas fragariae, Xanthomonas hortorum, X. melonis, Xanthomonas oryzae, Xanthomonas pisi, Xanthomonas vasicola and Xanthomonas vesicatoria. The species X. albilineans, X. sacchari, X. hyacinthi, X. theicola and X. translucens represented distinct clusters (II-VI). Topology of the 16S-23S rDNA ITS phylogenetic tree was very similar to that of the 16S rDNA tree reported previously, but more clusters were discriminated because of the higher level of diversity among the ITS sequences (16.2%) compared with the 16S rDNA sequences (1.8%). PMID- 11931145 TI - Comamonas koreensis sp. nov., a non-motile species from wetland in Woopo, Korea. AB - A bacterial strain, designated YH12T, was isolated from a wetland sample collected from Woopo, Republic of Korea, and characterized using a polyphasic approach. Analysis of 16S rDNA indicated that the isolate formed a monophyletic clade with the members of the genus Comamonas. The closest phylogenetic relative among the valid species was Comamonas testosteroni, with 96.6% 16S rDNA similarity. The chemotaxonomic properties of the wetland isolate supported its membership of the genus Comamonas, as it contained ubiquinone Q-8 as a major respiratory quinone and hexadecanoic, methylene-hexadecanoic and octadecenoic acids as major cellular fatty acids. The G+C content of the DNA was 66 mol%. The isolate is a gram-negative, non-pigmented, rod-shaped, oxidase- and catalase positive, non-motile, non-endospore-forming and non-fermentative bacterium. The phenotypic properties of the isolate were compared with those of the type strains of Comamonas terrigena, C. testosteroni and Delftia acidovorans. A number of tests, including motility, can differentiate our isolate from related taxa. On the basis of the 16S rDNA phylogenetic, chemotaxonomic and phenotypic evidence given in this study, it is proposed that strain YH12T (= KCTC 12005T = IMSNU 11158T) be assigned as the type strain of a novel species of the genus Comamonas, Comamonas koreensis sp. nov. PMID- 11931144 TI - Pseudomonas mosselii sp. nov., a novel species isolated from clinical specimens. AB - Twenty-two fluorescent pseudomonad strains of clinical origin received as Pseudomonas fluorescens (10 strains), Pseudomonasputida (10 strains) and Pseudomonas sp. (2 strains), and 33 type strains of the genus Pseudomonas were studied by numerical analysis based on 280 phenotypic characters. Twelve of the 22 clinical isolates clustered within a specific group, cluster IV. The other strains clustered within groups containing well-characterized fluorescent Pseudomonas species or did not cluster. Strains belonging to cluster IV were phenotypically different from all other clusters and subclusters of fluorescent pseudomonads. DNA-DNA hybridization showed that cluster IV corresponded to a genomic group sharing 72-100% DNA relatedness. DNA-DNA hybridization values with 67 strains representing 30 species of the genus Pseudomonas sensu stricto, including six recently described species (Pseudomonas veronii, Pseudomonas rhodesiae, Pseudomonas libanensis, 'Pseudomonas orientalis', 'Pseudomonas cedrella' and Pseudomonas monteilii), were below 49%, the value found for P. monteilii. The DNA G+C content of the type strain was 63 mol%. Comparison of the 16S rRNA gene sequence of a representative strain of cluster IV (CFML 90-83T) with sequences of other strains of the genus Pseudomonas revealed that strain CFML 90-83T was part of the P. fluorescens intrageneric cluster. On the basis of phenotypic, DNA-DNA hybridization and phylogenetic analyses, a novel species, Pseudomonas mosselii sp. nov., is proposed for the 12 strains of cluster IV. The type strain is P. mosselii CFML 90-83T (= ATCC BAA-99T = CIP 105259T). The P. mosselii strains are phenotypically homogeneous and can be differentiated from other fluorescent species by several phenotypic features, including pyoverdine typing. PMID- 11931146 TI - Bartonella bovis Bermond et al. sp. nov. and Bartonella capreoli sp. nov., isolated from European ruminants. AB - Two novel species of Bartonella isolated from European ruminants are described. Bartonella capreoli sp. nov. was isolated from the blood of roe-deer (Capreolus capreolus) captured in Chize, France. The type strain is IBS 193T (= CIP 106691T = CCUG 43827T). It is distinct from another European ruminant isolate that originated from a cow from a French herd of 430 dairy cattle. The latter isolate belongs to a novel species named Bartonella bovis Bermond et al. sp. nov. The type strain is strain 91-4T (= CIP 106692T = CCUG 43828T). The two bacteria appeared as small, fastidious, aerobic, oxidase-negative, gram-negative rods. Their biochemical properties were similar to those of members of the genus Bartonella. The sequences of the 16S rRNA and citrate synthase genes obtained from the two type strains were highly related to sequences of the different Bartonella species. Hybridization values when testing type strains of recognized Bartonella species, obtained with the nuclease/trichloroacetic acid method, support the creation of two novel species. PMID- 11931147 TI - Desulfotomaculum thermobenzoicum subsp. thermosyntrophicum subsp. nov., a thermophilic, syntrophic, propionate-oxidizing, spore-forming bacterium. AB - From granular sludge from a laboratory-scale upflow anaerobic sludge bed reactor operated at 55 degrees C with a mixture of volatile fatty acids as feed, a novel anaerobic, moderately thermophilic, syntrophic, spore-forming bacterium, strain TPO, was enriched on propionate in co-culture with Methanobacterium thermoautotrophicum Z245. The axenic culture was obtained by using pyruvate as the sole source of carbon and energy. The cells were straight rods with pointed ends and became lens-shaped when sporulation started. The cells were slightly motile. The optimum growth temperature was 55 degrees C and growth was possible between 45 and 62 degrees C. The pH range for growth of strain TPO was 6-8, with an optimum at pH 7-7.5. Propionate was converted to acetate, CO2 and CH4 by a co culture of strain TPO with Methanobacterium thermoautotrophicum Z245. In pure culture, strain TPO could grow fermentatively on benzoate, fumarate, H2/CO2, pyruvate and lactate. Sulphate could serve as inorganic electron acceptor when strain TPO was grown on propionate, lactate, pyruvate and H2/CO2. The G+C content was 53.7 mol%. Comparison of 16S rDNA sequences revealed that strain TPO is related to Desulfotomaculum thermobenzoicum (98%) and Desulfotomaculum thermoacetoxidans (98%). DNA-DNA hybridization revealed 88.2% reassociation between strain TPO and D. thermobenzoicum and 83.8% between strain TPO and D. thermoacetoxidans. However, both organisms differ physiologically from strain TPO and are not capable of syntrophic propionate oxidation. It is proposed that strain TPO should be classified as new subspecies of D. thermobenzoicum as D. thermobenzoicum subsp. thermosyntrophicum. PMID- 11931148 TI - Gelria glutamica gen. nov., sp. nov., a thermophilic, obligately syntrophic, glutamate-degrading anaerobe. AB - A novel anaerobic, gram-positive, thermophilic, spore-forming, obligately syntrophic, glutamate-degrading bacterium, strain TGO(T), was isolated from a propionate-oxidizing methanogenic enrichment culture. The axenic culture was obtained by growing the bacterium on pyruvate. Cells were rod-shaped and non motile. The optimal temperature for growth was 50-55 degrees C and growth occurred between 37 and 60 degrees C. The pH range for growth was 5.5-8 with optimum growth at pH 7. In pure culture, strain TGO(T) could grow on pyruvate, lactate, glycerol and several sugars. In co-culture with the hydrogenotrophic methanogen Methanobacterium thermautotrophicum strain Z-245, strain TGO(T) could grow on glutamate, proline and Casamino acids. Glutamate was converted to H2, CO2, propionate and traces of succinate. Strain TGO(T) was not able to utilize sulphate, sulphite, thiosulphate, nitrate or fumarate as electron acceptors. The G+C content was 33.8 mol%. Sequence analysis of the 16S rDNA revealed that strain TGO(T) belongs to the thermophilic, endospore-forming anaerobes, though no close relations were found. Its closest relations were Moorella glycerini (92%) and Moorella thermoacetica (90%). Strain TGOT had an unusually long 16S rDNA of more than 1700 bp. The additional base pairs were found as long loops in the V1, V7 and V9 regions of the 16S rDNA. However, the loops were not found in the 16S rRNA. The name Gelria glutamica gen. nov., sp. nov. is proposed for strain TGO(T). PMID- 11931149 TI - Rhodococcus jostii sp. nov., isolated from a medieval grave. AB - The taxonomic position of a bacterial strain isolated from the femur of the remains of Jost Lucembursky, margrave in Moravia, Brno (Czech Republic), was investigated by phenotypic, chemotaxonomic and molecular taxonomic methods. The chemotaxonomic characteristics, including the cell-wall amino acid and sugar compositions, the quinone system and the fatty acid profile, were in good agreement with those of the genus Rhodococcus. The G+C content of the DNA was 67.4 mol%. Comparative 16S rRNA gene sequencing demonstrated that the unknown strain represents a distinct line of descent within the genus Rhodococcus. The nearest relatives of the bacterium were Rhodococcus opacus and Rhodococcus percolatus. The unknown bacterium was readily distinguished from these species by using phenotypic methods. On the basis of phylogenetic and phenotypic evidence, it is proposed that the unknown bacterium be classified as Rhodococcus jostii sp. nov. The type strain is strain IFO 16295T (= CCM 4760T). PMID- 11931150 TI - Paenibacillus chinjuensis sp. nov., a novel exopolysaccharide-producing bacterium. AB - A novel exopolysaccharide-producing bacterium (WN9T) was isolated from Chinju, Korea, and was identified as a member of the genus Paenibacillus on the basis of phenotypic characteristics and phylogenetic inference based on 16S rDNA sequence. This organism is a facultatively anaerobic, endospore-forming rod. The diamino acid found in the peptidoglycan is meso-diaminopimelic acid. The predominant menaquinone is MK-7. The major cellular fatty acid is anteiso-C15:0. The G+C content is 53 mol%. The phylogenetic tree shows that strain WN9T falls within the radiation of a cluster comprising the Paenibacillus species. The levels of 16S rDNA similarity between strain WN9T and the type strains of validly described Paenibacillus species are 92.1-95.8%. Strain WN9T is clearly distinguishable from some phylogenetically related Paenibacillus species on the basis of DNA-DNA relatedness data and phenotypic characters. Therefore, on the basis of these data, a novel species of the genus Paenibacillus, Paenibacillus chinjuensis sp. nov., is proposed. The type strain is strain WN9T (= KCTC 8951PT = JCM 10939T). PMID- 11931151 TI - Reclassification of Eubacterium formicigenerans Holdeman and Moore 1974 as Dorea formicigenerans gen. nov., comb. nov., and description of Dorea longicatena sp. nov., isolated from human faeces. AB - Two strains of a gram-positively staining, obligately anaerobic, non-spore forming, rod-shaped bacterium, designated strains 111-13A and 111-35T, were isolated from human faeces. Analysis of the 16S rRNA gene sequences indicated that these strains were members of the Clostridium coccoides rRNA group of organisms. The nearest relatives of the unknown bacterium were Eubacterium formicigenerans (having a sequence similarity of 94%) and an uncultured bacterium (similarity > 99%). Characterization studies indicated that the unidentified faecal bacterium was biochemically distinct from Eubacterium formicigenerans, members of the Clostridium coccoides group and all other described Eubacterium species. On the basis of the data from these studies, it is proposed that the hitherto unknown rod-shaped bacterium be designated a species of a novel genus, namely Dorea longicatena gen. nov., sp. nov., and that Eubacterium formicigenerans be transferred to this genus as Dorea formicigenerans gen. nov., comb. nov. PMID- 11931152 TI - Ilyobacter insuetus sp. nov., a fermentative bacterium specialized in the degradation of hydroaromatic compounds. AB - The mesophilic, anaerobic bacterium strain VenChi2T was isolated with quinic acid (1,3,4,5-tetrahydroxy-cyclohexane-1-carboxylic acid) as the sole source of carbon and energy. Of more than 30 substrates tested, only quinic acid and shikimic acid (3,4,5-trihydroxy-1-cyclohexene-1-carboxylic acid) were utilized, yielding acetate, propionate, butyrate, H2 and CO2 as fermentation products. Sugars, alcohols, (di-)carboxylic acids, amino acids and aromatic compounds were not fermented and no external electron acceptors were used. Strain VenChi2T is a gram negative, strictly anaerobic, coccoid rod; it does not employ the classical hydroaromatic pathway of aerobic bacteria for the degradation of hydroaromatic compounds (no aromatic intermediates involved). Comparative 16S and 23S rDNA sequence analyses placed strain VenChi2T in the fusobacteria phylum, with the closest relatives among species of the genera Ilyobacter and Propionigenium. The results indicate that, disregarding the taxonomically misplaced Ilyobacter delafieldii, which is a member of the clostridia, the validly described Ilyobacter and Propionigenium species are phylogenetically intermixed. Based on its phenotypic properties, strain VenChi2T (= DSM 6831T = ATCC BAA-291T) is assigned to the genus Ilyobacter as the type strain of a novel species, Ilyobacter insuetus sp. nov. PMID- 11931153 TI - Biochemical and genetic evidence for the transfer of Mycobacterium tuberculosis subsp. caprae Aranaz et al. 1999 to the species Mycobacterium bovis Karlson and Lessel 1970 (approved lists 1980) as Mycobacterium bovis subsp. caprae comb. nov. AB - We propose to replace the species designation Mycobacterium tuberculosis subsp. caprae (Aranaz et al. 1999) by Mycobacterium bovis subsp. caprae comb. nov., since isolates of this subspecies share their main growth, biochemical and genetic characteristics with M. bovis and not with M. tuberculosis. These include negative biochemical test results for niacin accumulation and nitrate reduction as well as genetic features like the presence of an M. bovis-specific mutation in the oxyR locus, absence of the mtp40 sequence and a specific mutation in the gyrB gene, all of which have been described as characteristics for the differentiation of M. bovis. The only obvious biochemical character that differentiates the caprae subtype from other M. bovis isolates is susceptibility to pyrazinamide (PZA), which is due to the lack of a single point mutation in the pncA gene. However, susceptibility to PZA among clinical isolates of M. bovis isolates has been reported previously and, thus, may now been explained by a PZA-susceptible subspecies of M. bovis. We conclude that the species designation M. tuberculosis subsp. caprae is misleading and not correct in light of the biochemical and genetic characteristics and propose that the accurate designation of isolates of this subtype is M. bovis subsp. caprae. PMID- 11931154 TI - Helicobacter nemestrinae ATCC 49396T is a strain of Helicobacter pylori (Marshall et al. 1985) Goodwin et al. 1989, and Helicobacter nemestrinae Bronsdon et al. 1991 is therefore a junior heterotypic synonym of Helicobacter pylori. AB - Helicobacter nemestrinae Bronsdon et al. 1991, a gastric helicobacter species isolated from a pigtailed macaque, is thought to be the species most closely related to the important human pathogen Helicobacter pylori. The only available strain of this taxon is the type strain, ATCC 49396T. We sequenced seven housekeeping genes and two flagellin genes for H. nemestrinae ATCC 49396T. If ATCC 49396T were a separate species, these sequences should have been distinct from those of H. pylori. Instead, all sequences clustered together with sequences obtained previously for 20 or more H. pylori isolates from diverse geographical locations. The 16S rDNA sequence differed from that reported previously for this strain by 38 nucleotides and was most similar to that of H. pylori 85D08 (accession no. U00769), which was isolated from a rhesus macaque. It differed by less than 1% from 16S rDNA sequences of numerous other H. pylori strains, including the type strain, NCTC 11637T (= ATCC 43504T). These data indicate that the strain currently distributed as H. nemestrinae ATCC 49396T is really a strain of H. pylori and that H. nemestrinae Bronsdon et al. 1991 is a junior heterotypic synonym of Helicobacter pylori (Marshall et al. 1985) Goodwin et al. 1989. PMID- 11931155 TI - Propionivibrio limicola sp. nov., a fermentative bacterium specialized in the degradation of hydroaromatic compounds, reclassification of Propionibacter pelophilus as Propionivibrio pelophilus comb. nov. and amended description of the genus Propionivibrio. AB - Strain GolChi1T, a mesophilic, anaerobic bacterium, was isolated with quinic acid (1,3,4,5-tetrahydroxy-cyclohexane-1-carboxylic acid) as the sole source of carbon and energy. Of more than 30 substrates tested, only the hydroaromatic compounds quinic acid and shikimic acid (3,4,5-trihydroxy-1-cyclohexene-1-carboxylic acid) were utilized, yielding acetate and propionate as the only fermentation products. Sugars, alcohols, (di-)carboxylic acids, amino acids and aromatic compounds were not fermented and no external electron acceptors were used. Strain GolChi1T is a gram-negative, rod-shaped, aerotolerant anaerobe that possesses superoxide dismutase; it does not employ the classical hydroaromatic pathway of aerobic bacteria for the degradation of hydroaromatic compounds (no aromatic intermediates involved). 16S-rRNA-based phylogenetic analyses revealed a common origin of this isolate and Rhodocyclus, Propionibacter and Propionivibrio species. High sequence similarity (> 96%) and phenotypic traits indicated a closer relationship between strain GolChi1T and the type species of the monospecific genera Propionivibrio and Propionibacter but, due to its phenotypic properties, strain GolChi1T could not be assigned conclusively to either of these taxa. We propose (i) the amended description of the genus Propionivibrio, (ii) the reclassification of Propionibacter pelophilus Meijer et al. 1999 as Propionivibrio pelophilus comb. nov. and (iii) designation of Propionivibrio limicola sp. nov., with the type strain GolChi1T (= DSM 6832T = ATCC BAA-290T). PMID- 11931156 TI - Classification of three airborne bacteria and proposal of Hymenobacter aerophilus sp. nov. AB - Three aerobic, gram-negative, rod-shaped, non-spore-forming, red-pigmented, airborne bacteria (I/26-Cor1T, I/32A-Cor1 and I/74-Cor2) collected in the Museo Correr (Venice, Italy) were investigated to determine their taxonomic status by analysing their biochemical, physiological and chemotaxonomic features and the G+C content of genomic DNA and by comparing their genomic fingerprints. Additionally, the almost complete 16S rRNA gene sequence of strain I/26-Cor1T was analysed. The three strains were nearly identical in their morphological, physiological, biochemical and chemotaxonomic properties. The strains contained a menaquinone system with the predominant menaquinone MK-7 and a fatty acid profile with C15:0 anteiso, C15:0 iso and C16:1 predominant. Phosphatidylethanolamine and several unidentified lipids were detected in the polar lipid profiles. The polyamine pattern consisted of sym-homospermidine as the major compound. meso Diaminopimelic acid was found as the characteristic cell-wall diamino acid. The DNA base composition of the three strains ranged from 60 to 63 mol% G+C. Phylogenetically, strain I/26-Cor1T was most closely related to Hymenobacter actinosclerus (95.8% 16S rRNA gene sequence similarity). Physiological and genomic characteristics indicated that the two strains I/26-Cor1T and I/32A-Cor1 are representatives of the same species. The phylogenetic distance to any validly described taxon as indicated by 16S rRNA gene sequence similarities demonstrates that I/26-Cor1T and I/32A-Cor1 represent a novel species, for which the name Hymenobacter aerophilus sp. nov. is proposed, with the type strain I/26-Cor1T (= DSM 13606T = LMG 19657T). I/32A-Cor1 (= DSM 13607 = LMG 19658) is another strain of the species Hymenobacter aerophilus. Since the taxonomic status of strain I/74 Cor2 within the genus Hymenobacter was not determined unambiguously, it is designated Hymenobacter sp. I/74-Cor2 (= DSM 13611 = LMG 19659). PMID- 11931157 TI - Identification of isolates from soybean nodules in Xinjiang Region as Sinorhizobium xinjiangense and genetic differentiation of S. xinjiangense from Sinorhizobium fredii. AB - Eight fast-growing rhizobial isolates from Xinjiang soils were identified as Sinorhizobium xinjiangense by analyses of 16S rRNA gene sequences, SDS-PAGE of proteins, intergenic spacer sequences and DNA-DNA hybridization. Based on all of the results, these isolates and the reference strains for S. xinjiangense were a distinct genomic species, although the 16S rRNA genes were closely related to that of Sinorhizobium fredii. PMID- 11931158 TI - Species distinction of the ascomycetous heterothallic yeast-like fungus Stephanoascus ciferrii complex: description of Candida allociferrii sp. nov. and reinstatement of Candida mucifera Kockova-Kratochvilova et Slavikova. AB - The nucleotide sequences of the 18S rRNA gene (rDNA) from nine strains of the heterothallic ascomycetous Stephanoascus ciferrii complex were determined and the strains were separated into three groups according to their sequences. 18S rDNA sequences were identical within the same group. In group A the 18S rDNA sequences had no introns; in group B there was one group I intron, Sc1506-1 at position 1506; and in group C there were two group I introns, Sc943 at position 943 and Sc1506-2 at position 1506. Sc1506-1 and Sc1506-2 at position 1506 exhibited 19 base differences but were very similar. Therefore, it is suggested that these introns existed in the common ancestor of groups B and C, and that they were vertically inherited. DNA similarity values showed that the strains within the same group were of identical species. Group B included the isotype strains of Stephanoascus ciferrii and the type strains of Candida ciferrii and Sporothrix catenata; this confirmed that group B strains correspond to Stephanoascus ciferrii and that Candida ciferrii and Sporothrix catenata are synonyms of Stephanoascus ciferrii. The single member of group C, strain IFO 10918T, corresponds to the type strain of Candida mucifera and was independent of the other tested strains. Thus, Candida mucifera should be regarded as an independent species from Stephanoascus ciferrii. It is suggested that group A strains might comprise a new Stephanoascus species, but since group A strains could not form asci by themselves in this study they are described as a new species for which the name Candida allociferrii sp. nov. (type strain IFO 10194T) is proposed. PMID- 11931159 TI - Thermomonas haemolytica gen. nov., sp. nov., a gamma-proteobacterium from kaolin slurry. AB - Four aerobic, gram-negative bacterial strains isolated from kaolin slurry used in the production of paper were subjected to a polyphasic analysis and characterization to determine their taxonomic position. Analysis of the 16S rDNA sequences of the four strains revealed that they represent a new lineage within the gamma-Proteobacteria, related to the genera Xanthomonas, Pseudoxanthomonas, Stenotrophomonas, Luteimonas, Xylella and Rhodanobacter. Analysis of the quinone system, the polyamines, the fatty acids and the polar lipids revealed a combination of characteristics that is unique and not described for the phylogenetic relatives. The four strains contain a ubiquinone Q-8, spermidine as the major polyamine, diphosphatidylglycerol, phosphatidylglycerol and phosphatidylethanolamine as the predominant polar lipids, and a fatty acid profile with predominantly iso-branched fatty acids. The G+C content of the genomic DNA was determined to be within the narrow range 67.1-68.7 mol%. Determination of DNA relatedness, as well as riboprint band patterns and amplified fragment length polymorphism profiles, clearly demonstrated that the four strains are members of a single species. Antibiotic-susceptibility patterns were identical for the four strains. Although showing a high degree of similarites in physiological and biochemical patterns, each of the four strains could be distinguished from the others on the basis of a few biochemical characteristics. On the basis of the estimates of phylogenetic relationships derived from the 16S rDNA sequence analyses, the observed chemotaxonomic characteristics and other phenotypic traits, a new genus, Thermomonas gen. nov., and species, Thermomonas haemolytica sp. nov., are proposed for the strains A50-7 3T (= DSM 13605T = LMG 19653T), B 50-7-1 (= DSM 13598 = LMG 19655), D50-7-1 (= DSM 13610 = LMG 19656) and B50-8-1 (= DSM 13599 = LMG 19654), with strain A50-7 3T as the type strain. PMID- 11931160 TI - Salinibacter ruber gen. nov., sp. nov., a novel, extremely halophilic member of the Bacteria from saltern crystallizer ponds. AB - Five brightly red-pigmented, motile, rod-shaped, extremely halophilic bacteria were isolated from saltern crystallizer ponds in Alicante (two strains) and Mallorca (three strains), Spain. They grew optimally at salt concentrations between 20 and 30% and did not grow below 15% salts. Thus, these isolates are among the most halophilic organisms known within the domain Bacteria. The temperature optimum was 37-47 degrees C. A single, yet to be identified pigment was present, with an absorption maximum at 482 nm and a shoulder at 506-510 nm. The G+C content of the DNA was 66.3-67.7 mol% and, together, they formed a homogeneous genomic group with DNA-DNA similarities above 70%. The 16S rRNA gene sequences were almost identical to sequences recovered earlier from the saltern biomass by amplification of bacterial small-subunit rRNA genes from DNA extracted from the environment. This phylotype, earlier described as 'Candidatus Salinibacter', was shown by fluorescence in situ hybridization to contribute between 5 and 25% of the prokaryote community of the saltern crystallizers. We have therefore succeeded in isolating a bacterium from the natural environment that, although being a major component of the community, was previously known by its phylotype only. Isolation of the organism now allows formal description of a novel genus and species, for which we propose the name Salinibacter ruber gen. nov., sp. nov. The type strain is strain M31T (= DSM 13855T = CECT 5946T). PMID- 11931161 TI - 16S-23S rDNA internal transcribed spacer sequences for analysis of the phylogenetic relationships among species of the genus Fusobacterium. AB - The 16S-23S rDNA internal transcribed spacer (ITS) regions of all currently defined Fusobacterium species and related taxa such as Leptotrichia buccalis, Sebaldella termitidis and Streptobacillus moniliformans, were analysed to examine inter- and intraspecies as well as subspecies relationships. For the ITS amplification, a new eubacterial universal primer pair was designed and used. The majority of the Fusobacterium strains, along with L. buccalis showed one major, and two to three weaker, distinct bands (short and long versions) with lengths of 800-830 bp and 1000-1100 bp. Nevertheless, six other patterns were also found within the genus Fusobacterium, demonstrating its heterogeneity. The ITS region was sequenced and found to consist both of conserved motifs, which functioned as a framework for alignment, and of variable sites, which provided high phylogenetic resolution. Analyses of the ITS-DNA sequences and ITS relative length (short version) allowed species and subspecies differentiation in most cases. The results confirmed the strikingly distant relationship between Fusobacterium prausnitzii and the genus Fusobacterium. Fusobacterium nucleatum subspecies, along with Fusobacterium naviforme, Fusobacterium simiae and Fusobacterium periodonticum, formed a cluster with an inherently high potential for diversification. Other clusters were formed by Fusobacterium necrophorum subspecies with Fusobacterium gonidaformans and by Fusobacterium varium with Fusobacterium mortiferum and Fusobacterium ulcerans. Fusobacterium russii as well as Fusobacterium perfoetens formed separate branches. Fusobacterium necrophorum subspp. necrophorum and funduliforme on the one hand, and Fusobacterium varium and Fusobacterium mortiferum on the other, were found to be very similar, even at the high-resolution ITS level. PMID- 11931162 TI - Bacillus pycnus sp. nov. and Bacillus neidei sp. nov., round-spored bacteria from soil. AB - Bacillus sphaericus sensu lato currently consists of seven or more groups of unrelated taxa, one of which is B. sphaericus sensu stricto and another of which is Bacillus fusiformis. Members of two groups (groups 6 and 7), in common with all other B. sphaericus-like organisms, are unable to grow anaerobically or to use common hexoses, pentoses and hexitols as sources of carbon, have G+C contents of 34-36 mol % and form round spores. Groups 6 and 7 can be differentiated from other B. sphaericus-like organisms by low DNA relatedness and by variations in whole-cell fatty acid composition. Unique characteristics of group 6 include the ability to oxidize beta-hydroxybutyrate, the non-requirement for biotin and thiamin and failure to grow in 5% NaCl. Distinctive traits of group 7 include the inability to oxidize pyruvate and a requirement for biotin, thiamin and cystine for growth. These data show that groups 6 and 7 represent two novel species, for which the names Bacillus pycnus sp. nov. and Bacillus neidei sp. nov., respectively, are proposed; the corresponding type strains are NRRL NRS-1691T (= JCM 11075T) and NRRL BD-87T (= JCM 11077T). PMID- 11931163 TI - Weissella kimchii sp. nov., a novel lactic acid bacterium from kimchi. AB - A gram-positive, catalase-negative, non-sporulating, facultatively anaerobic, short rod-shaped bacterium, with cells measuring 0.3-0.5 x 1-2 microm and designated strain CHJ3T, was isolated from partially fermented kimchi, a traditional Korean fermented vegetable food. The strain produced CO2 gas, D lactate from glucose and dextran from sucrose and hydrolysed aesculin and arginine. It also fermented N-acetylglucosamine, amygdalin, arbutin, cellobiose, D-fructose, galactose, beta-gentiobiose, gluconate, D-glucose, maltose, D mannose, salicin, sucrose and D-xylose. The G+C content of the DNA was 48.2 mol%. Phylogenetic analysis of 16S rRNA showed that strain CHJ3T is a member of the genus Weissella. The nearest phylogenetic relative of strain CHJ3T was Weissella confusa, with 16S rRNA similarity of 98.3%. However, strain CHJ3T could be differentiated from W. confusa on the basis of some phenotypic characteristics, analysis of whole-cell protein patterns and DNA-DNA hybridization data. These data suggest that strain CHJ3T be classified in the genus Weissella as a novel species, Weissella kimchii sp. nov. The type strain is CHJ3T (= KCCM 41287T = DSM 14295T = KCTC 3746T). PMID- 11931164 TI - Pseudomonas lini sp. nov., a novel species from bulk and rhizospheric soils. AB - The taxonomic position of eight fluorescent Pseudomonas strains isolated from bulk and rhizospheric soils, and from water was examined. These eight strains clustered in one phenon together with Pseudomomas mandelii (CFBP 4844T), but could still be differentiated from this type strain by four phenotypic features. The eight stains exhibited internal DNA-DNA hybridization values ranging from 60 to 100%, with deltaTm below 5 degrees C (3.9 and 4.3 degrees C) for the lowest values (60 and 66%). The percentages of hybridization with type or reference strains of other Pseudomonas species tested ranged from 12 to 60% (deltaTm = 5.5 degrees C), indicating that the eight isolates studied constituted a discrete DNA homology group. Comparison of the 16S rDNA sequence of the strain representing this group (CFBP 5737T) with the sequences of other strains belonging to the genus Pseudomonas revealed that strain CFBP 5737T was a member of this genus and that these bacteria did not cluster with any previously described species of the genus Pseudomonas. The eight isolates belonged to two siderovars different from those described so far. On the basis of the results of phenotypic, DNA-DNA and phylogenetic analyses, and of siderotyping, a new species, Pseudomonas lini sp. nov. (type strain CFBP 5737T) is proposed. PMID- 11931165 TI - Nocardiopsis halotolerans sp. nov., isolated from salt marsh soil in Kuwait. AB - A polyphasic taxonomic study of a halotolerant micro-organism, isolated from Kuwait salt marsh soil, revealed that this strain represents a novel Nocardiopsis species. The strain produced substrate and aerial mycelium, grew at 28-35 degrees C in salt concentrations of 0-15% and was slightly keratinolytic. Results of the 165 rDNA sequence comparison revealed that strain F100T clustered with strains of the genus Nocardiopsis. This is consistent with other data such as: (i) growth characteristics, i.e. the formation of a white to yellow aerial mycelium and the typical zig-zag form of hyphae, which fragment when ageing; (ii) the presence of DL-diaminopimelic acid and glucose plus ribose in whole-cell hydrolysates; (iii) the presence of phosphatidyl choline, phosphatidyl inositol, phosphatidyl glycerol, phosphatidyl methylethanolamine and diphosphatidyl glycerol in polar lipid extracts; (iv) the presence of menaquinones MK-10(H(0-6)) and MK-11(H(0-6)) in the non-polar fraction; (v) the presence of iso/anteiso-branched plus 10 methyl-branched fatty acids, showing the diagnostic combination for Nocardiopsis spp. of 14-methyl-hexadecanoic acid (18%), oleic acid (9%) and tuberculostearic acid (2%); and (v) the absence of mycolic acids. Analysis of 16S rDNA revealed that strain F100T represents a distinct taxon within Nocardiopsis. Based upon phenotypic differences to other members of the genus, a novel species, Nocardiopsis halotolerans sp. nov., is proposed. The type strain of the species is F100T (= DSM 44410T = NRRL B-24124T). PMID- 11931166 TI - Evolution of the gyrB gene and the molecular phylogeny of Enterobacteriaceae: a model molecule for molecular systematic studies. AB - Phylogenetic trees showing the evolutionary relatedness of Enterobacteriaceae based upon gyrB and 16S rRNA genes were compared. Congruence among trees of these molecules indicates that the genomes of these species are not completely mosaic and that molecular systematic studies can be carried out. Phylogenetic trees based on gyrB sequences appeared to be more reliable at determining relationships among Serratia species than trees based on 16S rRNA gene sequences. gyrB sequences from Serratia species formed a monophyletic group validated by significant bootstrap values. Serratia fonticola had the most deeply branching gyrB sequence in the Serratia monophyletic group, which was consistent with its atypical phenotypic characteristics. Klebsiella and Enterobacter genera seemed to be polyphyletic, but the branching patterns of gyrB and 16S rRNA gene trees were not congruent. Enterobacter aerogenes was grouped with Klebsiella pneumoniae on the gyrB phylogenetic tree, which supports that this species could be transferred to the Klebsiella genus. Unfortunately, 16S rRNA and gyrB phylogenetic trees gave conflicting evolutionary relationships for Citrobacter freundii because of its unusual gyrB evolutionary process. gyrB lateral gene transfer was suspected for Hafnia alvei. Saturation of gyrB genes was observed by the pairwise comparison of Proteus spp., Providencia alcalifaciens and Morganella morganii sequences. Depending on their level of variability, 16S rRNA gene sequences were useful for describing phylogenetic relationships between distantly related Enterobacteriaceae, whereas gyrB sequence comparison was useful for inferring intra- and some intergeneric relationships. PMID- 11931167 TI - Vibrio calviensis sp. nov., a halophilic, facultatively oligotrophic 0.2 microm fiIterabIe marine bacterium. AB - A gram-negative, facultatively anaerobic, straight to slightly curved rod-shaped bacterium (RE35F/12T) sensitive to vibriostatic agent O/129 was previously isolated from sea water (Western Mediterranean Sea, Bay of Calvi, Corsica, France) by 0.2 microm-membrane filtration. Strain RE35/F12T (= CIP 107077T = DSM 14347T) was facultatively oligotrophic, halophilic, required Na+ for growth and produced acid but no gas from D-glucose under anaerobic conditions. Comparative 165 rRNA gene-sequence analyses demonstrated that the bacterium is most closely related (94.3%) to Vibrio scophthalmi. Similarities to the sequences of all other established Vibrio species ranged from 93.6% (with Vibrio aestuarianus) to 90.7% (with Vibrio rumoiensis). Strain RE35/F12T occupies a distinct phylogenetic position; this is similar to the case of Vibrio hollisae, because RE35F/12T represents a relatively long subline of descent sharing a branching point with the outskirts species V. hollisae. The G+C content of the DNA was 49.5 mol%. Ubiquinone Q-8 was the main respiratory lipoquinone, and 16:1omega9cis, 16:0 and 18:1trans9, cis11 were the major cellular fatty acids, 16:1omega9cis being predominant. The polyamine pattern was characterized by the presence of the triamine sym-norspermidine. On the basis of the polyphasic information summarized above, a new Vibrio species is described for which the name Vibrio calviensis sp. nov. is proposed. PMID- 11931168 TI - Saccharomonospora halophila sp. nov., a novel halophilic actinomycete isolated from marsh soil in Kuwait. AB - An actinomycete, strain 8T, was isolated from marsh soil in Kuwait. The strain was aerobic, gram-positive, halophilic and produced light blue to greyish aerial mycelium. The warty spores were sessile, occurring singly or in pairs on aerial mycelium. The mycelium was stable and did not fragment during ageing. Chemotaxonomic markers of the isolate were consistent with its classification as Saccharomonospora. The strain possessed meso-diaminopimelic acid as the diagnostic amino acid in the peptidoglycan. The diagnostic sugars were arabinose and galactose; polar lipids were phosphatidyl inositol, phosphatidyl ethanolamine, hydroxy-phosphatidyl ethanolamine, lyso-phosphatidyl ethanolamine and diphosphatidyl glycerol; the principal menaquinone was MK-9(H4); and the iso/anteiso-branched fatty acid pattern was combined with 10-methyl-branched and 2-hydroxy-branched fatty acids. Saccharomonospora cyanea DSM 44106T was the closest phylogenetic neighbour of strain 8T, showing 96.8% 16S rDNA sequence similarity. These data, together with distinct physiological traits, led to the conclusion that the novel isolate represents a new species within the genus Saccharomonospora for which the name Saccharomonospora halophila sp. nov. is proposed. The type strain is strain 8T (= DSM 44411T =NRRL B-24125T). PMID- 11931169 TI - Stenotrophomonas acidaminiphila sp. nov., a strictly aerobic bacterium isolated from an upflow anaerobic sludge blanket (UASB) reactor. AB - Two of several strictly aerobic, mesophilic bacteria isolated from a lab-scale upflow anaerobic sludge blanket (UASB) reactor treating a petrochemical wastewater, strains AMX 17 and AMX 19T, were subjected to detailed taxonomic study. Cells were gram-negative, motile, non-sporulating, straight to curved rods with a polar flagellum. The isolates exhibited phenotypic traits of members of the genus Stenotrophomonas, including cellular fatty acid composition and the limited range of substrates that could be used. Sugars and many amino acids were utilized. Antibiotic susceptibility and physiological characteristics were determined. The DNA base composition was 66.9 mol% G+C. Phylogenetic analysis revealed that the nearest relatives were Stenotrophomonas maltophilia LMG 11114, Stenotrophomonas nitritireducens DSM 12575T and Pseudomonas pictorum ATCC 23328T (similarity of 98.1-98.8%). Xanthomonas species, S. maltophilia LMG 958T and Stenotrophomonas africana CIP 104854T showed high 16S rRNA sequence similarities (96.4-97.3%). The high similarity found in cellular fatty acid profiles and identical partial 16S rRNA sequences (500 bp) for strains AMX 17 and AMX 19T indicate that they belong to the same species. DNA-DNA hybridizations revealed respectively 26.7, 31, 65.8 and 43.6% homology between isolate AMX 19T and S. africana CIP 104854T, S. maltophilia CIP 60.77T, S. nitritireducens DSM 12575T and P. pictorum ATCC 23328T. These results allow the proposal of strain AMX 19T (= DSM 13117T = ATCC 700916T = CIP 106456T) as representative of a novel species of the genus Stenotrophomonas, with the name Stenotrophomonas acidaminiphila sp. nov. PMID- 11931170 TI - Arthrobacter nasiphocae sp. nov., from the common seal (Phoca vitulina). AB - An unknown gram-positive, catalase-positive, strictly aerobic, rod-shaped bacterium was isolated from the nasal cavities of two common seals. Chemical analysis revealed the presence in the bacterium of a hitherto unknown cell-wall murein [type: L-Lys-L-Ala2-Gly(2-3)-L-Ala (Gly)]. Comparative 16S rRNA gene sequencing showed that the unidentified rod was related to the Arthrobacter group of organisms, although sequence divergence values of >3% from established members of this genus indicated that it represents a novel species. On the basis of phenotypic and phylogenetic considerations, it is proposed that the unknown bacterium from seals (Phoca vitulina) be classified as a novel species, Arthrobacter nasiphocae sp. nov. The type strain of Arthrobacter nasiphocae is CCUG 42953T. PMID- 11931171 TI - A mathematical method for determining genome divergence and species delineation using AFLP. AB - The delineation of bacterial species is presently achieved using direct DNA-DNA relatedness studies of whole genomes. It would be helpful to obtain the same genomically based delineation by indirect methods, provided that descriptions of individual genome composition of bacterial genomes are obtained and included in species descriptions. The amplified fragment length polymorphism (AFLP) technique could provide the necessary data if the nucleotides involved in restriction and amplification are fundamental to the description of genomic divergences. Firstly, in order to verify that AFLP analysis permits a realistic exploration of bacterial genome composition, the strong correspondence between predicted and experimental AFLP data was demonstrated using Agrobacterium strain C58 as a model system. Secondly, a method is proposed for determining current genome mispairing and evolutionary genome divergences between pairs of bacteria, based on arbitrary sampling of genomes by using AFLP. The measure of current genome mispairing was validated by comparison with DNA-DNA relatedness data, which itself correlates with base mispairing. The evolutionary genome divergence is the estimated rate of nucleotide substitution that has occurred since the strains diverged from a common ancestor. Current genome mispairing and evolutionary genome divergence were used to compare members of Agrobacterium, used as a model of closely related genomic species. A strong and highly significant correlation was found between calculated genome mispairing and DNA-DNA relatedness values within genomic species. The canonical 70% DNA-DNA hybridization value used to delineate genomic species was found to correspond to a range of current genome mispairing of 13 13.6%. These limits correspond to 0.097 and 0.104 nucleotide substitutions per site, respectively. In addition, experimental data showed that the large Ti and cryptic plasmids of Agrobacterium had little effect on the estimation of genome divergence. Evolutionary genome divergence was used for phylogenetic inferences. Data showed that members of the same genomic species clustered consistently, as supported by bootstrap resampling. On the basis of these results, it is proposed that the genomic delineation of bacterial species could be based, in future, on phylogenetic groups supported by bootstraps and genome descriptions of individual strains, obtained by AFLP analysis, recorded in accessible databases; this approach might eventually replace DNA-DNA hybridization studies. PMID- 11931172 TI - Proposal of Ureaplasma parvum sp. nov. and emended description of Ureaplasma urealyticum (Shepard et al. 1974) Robertson et al. 2001. AB - The phenotypic and genotypic properties of Ureaplasma urealyticum (family Mycoplasmataceae, order Mycoplasmatales, class Mollicutes) are reviewed here. The 14 recognized serovar standard strains found in humans exhibit no serological cross-reactivity with ureaplasmas from other hosts and uniquely express human immuoglobulin A1 protease activity. However, they exhibit many characteristics which place them in two distinct clusters known as the parvo biovar (or biovar 1 or B) and the T960T biovar (or biovar 2 or A). Established phenotypic markers of the biovars include clustering of antigenic types, polypeptide patterns of whole cell preparations, differential inhibition by manganese, and polymorphism among their ureases, pyrophosphatases and diaphorases. Established genotypic markers of the biovars are DNA-DNA hybridization of 60% between biovars, and distinctive RFLP patterns and genome sizes. Divergent nucleotide sequences of several highly conserved genes attest to the phylogenetic distinctiveness of the two biovars. PCRs founded upon the sequences for 16S rRNA, the 16S-23S rRNA intergenic regions, the genus-defining urease, the serovar-defining, multiple-banded antigen genes or randomly amplified polymorphic DNA tests differentiate the biovars unambiguously. With the availability of rapid, reliable and economical tests for biovar determination, it is now appropriate to propose that the taxonomic status of U. urealyticum be emended. Serovar standard strains exhibiting traits of biovar parvo (serovars 1, 3, 6 and 14) will be designated as a separate species, Ureaplasma parvum sp. nov., as befits its smaller genome size. The serovar 3 standard (strain 27T) will be the type strain of U. parvum and is represented by ATCC 27815T and NCTC 11736T. Serovar standard strains exhibiting traits of biovar T960T (2, 4, 5, 7, 8T, 9, 10, 11, 12 and 13) will retain the U. urealyticum designation and type strain, the serovar 8 standard (strain T960T), represented by ATCC 27618T and NCTC 10177T. PMID- 11931173 TI - Obligate bacterial endosymbionts of Acanthamoeba spp. related to the beta Proteobacteria: proposal of 'Candidatus Procabacter acanthamoebae' gen. nov., sp. nov. AB - All obligate bacterial endosymbionts of free-living amoebae currently described are affiliated with the alpha-Proteobacteria, the Chlamydiales or the phylum Cytophaga-Flavobacterium-Bacteroides. Here, six rod-shaped gram-negative obligate bacterial endosymbionts of clinical and environmental isolates of Acanthamoeba spp. from the USA and Malaysia are reported. Comparative 16S rDNA sequence analysis demonstrated that these endosymbionts form a novel, monophyletic lineage within the beta-Proteobacteria, showing less than 90% sequence similarity to all other recognized members of this subclass. 23S rDNA sequence analysis of two symbionts confirmed this affiliation and revealed the presence of uncommon putative intervening sequences of 146 bp within helix-25 that shared no sequence homology to any other bacterial rDNA. In addition, the 23S rRNA of these endosymbionts displayed one polymorphism at the target site of oligonucleotide probe BET42a that is conserved in all other sequenced beta-Proteobacteria. Intra cytoplasmatic localization of the endosymbionts within the amoebal host cells was confirmed by electron microscopy and fluorescence in situ hybridization with a specific 16S rRNA-targeted oligonucleotide probe. Based on these findings, the provisional name 'Candidatus Procabacter acanthamoebae' is proposed for classification of a representative of the six endosymbionts of Acanthamoeba spp. studied in this report. Comparative 18S rDNA sequence analysis of the Acanthamoeba host cells revealed their membership with either Acanthamoeba 18S rDNA sequence type T5 (Acanthamoeba lenticulata) or sequence type T4, which comprises the majority of all Acanthamoeba isolates. PMID- 11931174 TI - Paenibacillus graminis sp. nov. and Paenibacillus odorifer sp. nov., isolated from plant roots, soil and food. AB - Sixteen gram-positive endospore-forming bacteria previously isolated from soil, plant rhizospheres, plant roots and pasteurized pureed vegetables were studied to determine their taxonomic positions. The isolates were formerly identified as Bacillus circulans based on their biochemical characters using API galleries. Two of these strains, RSA19T and TOD45T, were recently assigned to the genus Paenibacillus based on phylogenetic analysis of their 16S rRNA (rrs) gene sequence. In the present work, the sixteen isolates were assigned to two genomospecies using DNA-DNA hybridization, in agreement with rrs gene sequence analysis. These genomospecies can also be differentiated on the basis of their cultural and biochemical characters into two novel species, for which the names Paenibacillus graminis sp. nov. (type strain RSA19T = ATCC BAA-95T = LMG 19080T) and Paenibacillus odorifer sp. nov. (type strain TOD45T = ATCC BAA-93T = LMG 19079T) are proposed. PMID- 11931175 TI - Arcanobacterium hippocoleae sp. nov., from the vagina of a horse. AB - A polyphasic taxonomic study was performed on a previously unidentified gram positive, facultatively anaerobic, diphtheroid-shaped organism isolated from a vaginal discharge of a horse. Comparative 16S rRNA gene sequencing demonstrated that the strain was a member of the genus Arcanobacterium, but sequence divergence values of >4% with described species of this genus (viz: Arcanobacterium haemolyticum, Arcanobacterium bernardiae, Arcanobacterium phocae, Arcanobacterium pluranimalium and Arcanobacterium pyogenes) demonstrated that the isolate represented a novel species. The unknown bacterium was readily distinguished from other Arcanobacterium species by biochemical tests. Based on phylogenetic and phenotypic evidence, it is proposed that the unknown bacterium be classified as Arcanobacterium hippocoleae sp. nov. The type strain of A. hippocoleae is CCUG 44697T (= CIP 106850T). PMID- 11931176 TI - Nocardiopsis compostus sp. nov., from the atmosphere of a composting facility. AB - Three strains (KS8, KS9T and KS21), isolated from air samples near a composting facility, were subjected to taxonomic analyses (characterized using a polyphasic approach). Morphological and chemotaxonomic characteristics of the isolates were in agreement with those described for members of the genus Nocardiopsis. On the basis of 16S rRNA sequence comparison and phenotypic tests, KS21 clearly belonged to Nocardiopsis alba. KS8 and KS9T showed less than 98% 16S rRNA gene sequence similarity to any of the previously described Nocardiopsis species. The polar lipid profiles of both isolates consisted of four major compounds, phosphatidylmonomethylethanolamine, phosphatidylcholine, diphosphatidylglycerol and phosphatidylglycerol, in addition to two unknown phospholipids. The major menaquinones in KS8 and KS9T were MK-10(H8), MK-11(H8), MK-10(H6) and MK-12. Furthermore, MK-13, MK-11(H6), MK-9(H8) and MK-10(H4) could be detected in significant amounts. The fatty acid composition included iso- and anteiso branched acids combined with tuberculostearic acid (Me18:0), straight-chain saturated (16:0, 18:0) and unsaturated (16:1, 17:1, 18:1) fatty acids. On the basis of these results, KS8 and KS9T clearly represent a novel species of the genus Nocardiopsis, for which the name Nocardiopsis compostus sp. nov. is proposed (type strain KS9T = DSM 44551T= NRRL B-24145T). PMID- 11931177 TI - Emended descriptions of the genus Micrococcus, Micrococcus luteus (Cohn 1872) and Micrococcus lylae (Kloos et al. 1974). AB - Nine yellow-pigmented, spherical bacterial strains isolated from a medieval wall painting (strain D7), from indoor air (strains 3, 6, 7, 13C2, 38, 83 and 118) and from an activated-sludge plant (strain Ballarat) were classified by a polyphasic approach. Analyses of the 16S rRNA gene sequences of three representatives (strains D7, 118 and Ballarat) indicated that they all belong to the genus Micrococcus. The three isolates shared the highest sequence similarities with Micrococcus luteus DSM 20030T (97.9-98%), Micrococcus antarcticus AS 1.2372T (97.9-98.3%) and Micrococcus lylae DSM 20315T (97.5-97.9%). DNA-DNA reassociation studies clearly demonstrated that all nine isolates belong to the species M. luteus. However, neither their chemotaxonomic features nor their physiological and biochemical properties were consistent with those of M. luteus DSM 20030T. In contrast to M. luteus DSM 20030T, all isolates investigated possessed MK-8(H2) as the major respiratory quinone, and strain Ballarat had an A4alpha peptidoglycan type. On the basis of analyses of their Fourier transform-infrared spectroscopy spectra, isolates D7, 3, 6, 7, 13C2, 38, 83 and 118 could be grouped into a single cluster separate from M. luteus DSM 20030T, strain Ballarat and M. lylae DSM 20315T. In addition, all these isolates could be distinguished from M. luteus DSM 20030T by their ability to assimilate D-maltose, D-trehalose, DL-3 hydroxybutyrate, DL-lactate, pyruvate and L-histidine and to hydrolyse casein. Strains D7, 3, 6, 7, 13C2, 38, 83 and 118 differed from both M. luteus DSM 20030T and strain Ballarat by their ability to assimilate acetate, L-phenylalanine, L serine and phenylacetate. Furthermore, REP-PCR fingerprinting yielded one common band for these strains, whereas this band was not observed for M. luteus DSM 20030T, strain Ballarat or M. lylae DSM 20315T. On the basis of these data, the species M. luteus can be divided into three biovars that are distinguished by several chemotaxonomic and biochemical traits: biovar I, represented by M. luteus DSM 20030T; biovar II, represented by strains D7 (= DSM 14234 = CCM 4959), 3, 6, 7, 13C2, 38, 83 and 118; and biovar III, represented by strain Ballarat (= DSM 14235 = CCM 4960). On the basis of the results generated in this study, emended descriptions of the genus Micrococcus and the species M. luteus and M. lylae are given. PMID- 11931178 TI - Lactobacillus diolivorans sp. nov., a 1,2-propanediol-degrading bacterium isolated from aerobically stable maize silage. AB - Inoculation of maize silage with Lactobacillus buchneri (5 x 10(5) c.f.u. g(-1) of maize silage) prior to ensiling results in the formation of aerobically stable silage. After 9 months, lactic acid bacterium counts are approximately 10(10) c.f.u. g(-1) in these treated silages. An important subpopulation (5.9 x 10(7) c.f.u. g(-1)) is able to degrade 1,2-propanediol, a fermentation product of L. buchneri, under anoxic conditions to 1-propanol and propionic acid. From this group of 1,2-propanediol-fermenting, facultatively anaerobic, heterofermentative lactobacilli, two rod-shaped isolates were purified and characterized. Comparative 16S rDNA sequence analysis revealed that the newly isolated bacteria have identical 16S rDNA sequences and belong phylogenetically to the L. buchneri group. DNA-DNA hybridizations, whole-cell protein fingerprinting and examination of phenotypic properties indicated that these two isolates represent a novel species, for which the name Lactobacillus diolivorans sp. nov. is proposed. The type strain is LMG 19667T (= DSM 14421T). PMID- 11931179 TI - Leuconostoc ficulneum sp. nov., a novel lactic acid bacterium isolated from a ripe fig, and reclassification of Lactobacillus fructosus as Leuconostoc fructosum comb. nov. AB - An isolate, designated strain FS-1T, was recovered from a ripe fig. Phylogenetic analysis of the 16S rRNA genes and DNA-DNA reassociation values showed that the organism represented a novel species of the genus Leuconostoc closely related to Lactobacillus fructosus. The novel isolate could be distinguished from the type strain of Lactobacillus fructosus by the fatty acid composition and several phenotypic and growth characteristics. In strain FS-1T, 18:1 delta9 (18:1omega9c) was present in relatively large amounts whilst, in Lactobacillus fructosus, this fatty acid was a minor component. Strain FS-1T and Lactobacillus fructosus produced acid in API 50CHL microtubes from glucose, fructose and mannitol within 48 h, whereas only strain FS-1T also fermented trehalose, gluconate, turanose and sucrose after 48 h. Other differences in acid production from carbohydrates also distinguished strain FS-1T from Lactobacillus fructosus. Both organisms were heterofermentative with fructose as a substrate and fermented glucose only in the presence of fructose, as determined by nuclear magnetic resonance studies. Strain FS-1T was catalase-positive. On the basis of the phylogenetic analysis, DNA-DNA reassociation values, physiological and biochemical characteristics and fatty acid composition, the name Leuconostoc ficulneum is proposed for the novel species represented by strain FS-1T, and it is proposed that Lactobacillus fructosus be reclassified in the genus Leuconostoc as Leuconostoc fructosum comb. nov. PMID- 11931180 TI - Thioalkalivibrio thiocyanoxidans sp. nov. and Thioalkalivibrio paradoxus sp. nov., novel alkaliphilic, obligately autotrophic, sulfur-oxidizing bacteria capable of growth on thiocyanate, from soda lakes. AB - Nine strains of haloalkaliphilic, obligately autotrophic, sulfur-oxidizing bacteria able to grow with thiocyanate (SCN-) as the sole energy and nitrogen source were isolated from soda lakes in South-East Siberia, Kenya and Egypt after enrichment on sodium carbonate minerals buffered at pH 10 with thiocyanate as the substrate. The isolates fell into two groups that were substantially different in terms of cell morphology, growth parameters and the ability to oxidize carbon disulfide. The bacteria were able to oxidize sulfide, polysulfide, sulfur and tetrathionate, as well as thiocyanate. Two isolates belonged to an extremely halotolerant type growing in the presence of up to 4 M Na+. Cyanate (CNO-) was the main nitrogen-containing intermediate during thiocyanate degradation in both groups. According to DNA-DNA hybridization data and phylogenetic analysis, both groups of isolates belong to a recently described genus of haloalkaliphilic sulfur-oxidizing bacteria, i.e. Thioalkalivibrio, belonging to the gamma Proteobacteria, in which where they represent two new species. The species name Thioalkalivibrio paradoxus (type strain ARh 1T = DSM 13531T = JCM 11367T) is proposed for the group with barrel-shaped cells, and the species name Thioalkalivibrio thiocyanoxidans (type strain ARh 2T, DSM 13532T = JCM 11368T) is proposed for the group with vibrio-shaped cells. The diagnosis of the genus Thioalkalivibrio is amended according to the new data. PMID- 11931181 TI - Streptococcus entericus sp. nov., isolated from cattle intestine. AB - Biochemical, molecular chemical and molecular genetic studies were performed on an unknown gram-positive, catalase-negative, coccus-shaped organism isolated from the intestine of a cow affected with catarrhal enteritis. The organism was tentatively identified as a streptococcal species based on results of cellular morphological and biochemical tests. 16S rRNA gene sequencing studies confirmed its provisional identification as a member of the genus Streptococcus, but the organism did not correspond to any recognized species of this genus. The nearest phylogenetic relatives of the unknown coccus from a calf were Streptococcus acidominimus and Streptococcus suis. The unknown bacterium, however, was distinguished from these species and other animal streptococci by biochemical tests and electrophoretic analysis of whole-cell proteins. Based on both phenotypic and phylogenetic findings, it is proposed that the unknown bacterium be classified as a novel species of the genus Streptococcus, Streptococcus entericus sp. nov. The type strain is CECT 5353T (= CCUG 44616T). PMID- 11931182 TI - Thiobaca trueperi gen. nov., sp. nov., a phototrophic purple sulfur bacterium isolated from freshwater lake sediment. AB - Two strains of a novel species of phototrophic micro-organism were isolated from the sediments of a shallow, freshwater, eutrophic lake. Both strains grew photolithoheterotrophically with sulfide as an electron donor, transiently accumulating intracellular sulfur globules. Photolithoautotrophic growth was not observed. One strain was designated BCH(T) (the type strain) and was studied in most detail. Cells contained bacteriochlorophyll a, and the dominant carotenoid was lycopene. Cell suspensions were brown. The photosynthetic membranes had a vesicular arrangement. Acetate, propionate, pyruvate, succinate and fumarate were each used as electron donors and carbon sources in the presence of sulfide and bicarbonate. In the presence of light, growth did not occur with hydrogen, thiosulfate or iron(II). The optimum temperature for growth was between 25 and 30 degrees C, the maximum being 36 degrees C. The G+C content of the genomic DNA of strain BCH(T) was 63 mol%. Analysis of the 16S RNA genes showed that both strains belonged to the gamma-subclass of the Proteobacteria but were phylogenetically distinct from any described phototrophic organisms within the Chromatiaceae. On the basis of phylogenetic and physiological differences from other phototrophic microorganisms, strain BCH(T) is described as a novel species of a new genus, Thiobaca trueperi gen. nov., sp. nov. PMID- 11931183 TI - Emended description of Paracoccus kondratievae. AB - An aerobic, facultatively chemolithotrophic and methylotrophic strain, GB, was isolated from a maize rhizosphere. On the basis of comparative analysis of its phenotypic and genotypic properties, it is proposed that strain GB(T) (= VKM B 2222T = NCIMB 13773T) be assigned to the genus Paracoccus as Paracoccus kondratievae sp. nov. PMID- 11931184 TI - Revision of haemotrophic Mycoplasma species names. AB - The recently proposed transfer of four rickettsias from the genera Haemobartonella and Eperythrozoon to the genus Mycoplasma with the Candidatus status is herein revised. This is because the Candidatus designation is for new, incompletely described taxa, in order to give them a provisional status. Thus, 'Candidatus Mycoplasma haemofelis' is revised to Mycoplasma haemofelis comb. nov., nom. nov., 'Candidatus Mycoplasma haemomuris' is revised to Mycoplasma haemomuris comb. nov., nom. nov., 'Candidatus Mycoplasma haemosuis' is revised to Mycoplasma haemosuis comb. nov., nom. nov. and 'Candidatus Mycoplasma wenyonii' is revised to Mycoplasma wenyonii comb. nov. PMID- 11931185 TI - The use of razor blades, nail clippers and other tools for nail cutting. PMID- 11931186 TI - Complications of gynaecological endoscopic surgery at King Edward VIII Hospital, Durban, South Africa. PMID- 11931187 TI - Visual loss in urban and rural chronic glaucoma patients in Ghana. AB - The visual acuity of newly diagnosed glaucoma patients in a rural and an urban area were compared in a hospital-based cross-sectional study in Ghana. Age at presentation was strikingly higher in the rural group due to local factors which result in a difference-in-age structure of the rural and urban population. The crude relative frequency of bilateral blindness was significantly higher for rural than urban. However, with age adjusted relative frequencies for bilateral blindness there was no significant difference between rural and urban patients. There was significant difference between them for those aged 51-71 when blindness in one or both eyes were considered together. There was more consistent blindness in the rural community: the difference was attributed to local factors such as better access to eye care in the urban group. PMID- 11931188 TI - A personal view from Cambodia. PMID- 11931189 TI - Aid and reform in Africa: lessons from 10 case studies: final report. PMID- 11931190 TI - Heterotopic twin delivery of live infants. PMID- 11931191 TI - Severe chemical proctitis following application of caustic native suppository. PMID- 11931192 TI - An unusual cyst in the broad ligament. PMID- 11931193 TI - Co-existent tuberculosis and papillary carcinoma thyroid. PMID- 11931194 TI - Childhood tuberculosis in a rural tropical area: risk factors. PMID- 11931195 TI - Cysticercosis masquerading as a salivary gland neoplasm. PMID- 11931196 TI - Chemotherapeutic failures in Plasmodium falciparum infections. PMID- 11931197 TI - Gall bladder cancer risk factors and genetic mutations in Pakistani women. PMID- 11931198 TI - HIV in Nepal: times have changed. PMID- 11931199 TI - Burns management. PMID- 11931200 TI - The Roll Back Malaria movement. PMID- 11931201 TI - Structural adjustment and health sector reform in Africa. PMID- 11931202 TI - Glaucoma in the developing world. PMID- 11931203 TI - Thrombolytic therapy in acute myocardial infarction in a rural setting. PMID- 11931204 TI - Epidemic dropsy: observations on pathophysiology and clinical features during the Delhi epidemic of 1998. AB - Epidemic dropsy results from the consumption of edible oils adulterated with Argemone mexicana oil by unscrupulous traders. Twenty consecutive 'in-door' patients of dropsy were intensively studied during the recent Delhi epidemic. Samples of edible oil used by them, their urine and their serum samples tested positive for sanguinarine on thin layer chromatography. The illness starts as a gastro-enteric illness followed by oliguria and pedal oedema. The following are often observed: cutaneous erythema with blanching and tenderness on pressure; violacious pigmentation of the skin; shortness of breath with orthopnoea; right sided heart failure with normal left ventricle (LV) functions; as well as severe anaemia and hypoalbuminaemia. Renal function tests showed: bland urinary sediments; decreased glomerular filtration rate (GFR); mild to moderate azotaemia; acute tubular necrosis; patchy pneumonitis; moderate hypoxia with respiratory alkalosis; and restrictive ventilatory defects on blood gas analysis; and spirometry suggestive of interstitial pulmonary oedema of non-cardiogenic origin. 99mTc colloid sulphur liver scans showed colloid shift. There was marked dilatation and proliferation of dermal capillaries in the absence of significant inflammation in the biopsy specimens. Toxic alkaloids of Argemone mexicana oil induce widespread capillary dilatation and permeability causing leakage of protein rich plasma into the interstitial tissues of various organs. A hypovolaemic state is thus induced producing renal hypoperfusion which may progress to acute tubular necrosis. Interstitial fluid in alveoli causes restrictive ventilatory dysfunction with hypertension and right-sided failure with well-preserved LV function. The hepatic venous congestion induces Kupffer's cell dysfunction, which results in colloid shift on a radionuclide liver scan. PMID- 11931205 TI - Referral to hospital in Nepal: 4 years' experience in one rural district. AB - Formal referral systems have been proposed as a strategy to improve access to secondary care, yet their implementation can be problematic. This paper describes data from referrals in one rural district in Nepal over a 4-year period. Whilst the characteristics of those patients attending hospital after referral were similar to those described in other developing countries, the rate (1.0/1000 population/year) is much lower, especially when compared to estimated need. Geographical and other barriers to access to secondary care in rural Nepal are discussed. PMID- 11931206 TI - Sigmoid volvulus in rural Bengal. AB - Between January 1995 and December 2000, 197 patients (44.97%) were treated for sigmoid volvulus at Bankura Medical College and Hospital, Bankura, West Bengal, India. The mean age was 45.06 years and the male to female ratio was 2.07:1. The various aetiological factors, clinical features and management have been reviewed. Abdominal distension with obstipation was the common clinical presentation. Straight X-ray of the abdomen suggested the diagnosis. Management included celiotomy and primary resection and anastomosis without intra-operative lavage in 196 patients and sigmoidopexy in one patient. The overall mortality rate was 1.01%. There was only one recurrence in the patients who had undergone sigmoidopexy. Primary resection and anastomosis at initial presentation proved a safe operative treatment and avoided unnecessary repeated hospitalization. PMID- 11931207 TI - Infective endocarditis: report of a prospective study in an Indian hospital. AB - Forty-six patients who fulfilled the Duke's clinical diagnostic criteria for infective endocarditis (IE) were evaluated. Thirty-five (76%) patients were below 40 years of age with rheumatic heart disease being the most common underlying heart lesion affecting 26 (56%). An obvious predisposing cardiac lesion could not be ascertained in 22%. Blood culture positivity was 44% with streptococcus heading the list. The incidence of the staphylococcal (25%) and gram negative bacillary endocarditis (15%) were found to be increasing. Streptococci were susceptible to penicillin with minimum bactericidal concentration: minimum inhibitory concentration within acceptable limits. However, the appearance of methicillin resistant staphylococcus aureus and high level gentamicin resistant enterococcus as aetiological agents of infective endocarditis were found to add to the complexity of the problem. With the emergence of drug-resistant organisms as causative agents of IE, whenever medical therapy is the primary method of treatment of this condition, the selection of antibiotics should depend upon extensive in vitro testing and in vivo monitoring of clinical efficacy. PMID- 11931208 TI - Cleft lip and palate. PMID- 11931209 TI - Childhood brucellosis is still a severe problem in the eastern region of Turkey. PMID- 11931210 TI - Clinical manifestations and ultrasonography guided aspiration of amoebic liver abscesses in St Walburg's Hospital, Nyangao, Tanzania. PMID- 11931211 TI - Broad band ultraviolet light, midrange sunbeam spectrum in the treatment of vitiligo. PMID- 11931212 TI - Effectiveness of a health education intervention in a war environment. PMID- 11931213 TI - Abdominal tuberculosis adenitis in HIV-infected patients: is ultrasound diagnosis appropriate? PMID- 11931215 TI - Trichobezoar obstructing the terminal ileum. PMID- 11931214 TI - The resistible rise of surgical sepsis in Malawi. PMID- 11931216 TI - The burden of restorative dental treatment for children in Third World countries. AB - OBJECTIVE: To analyse whether developing countries have sufficient health dollars to treat existing diseases in general and dental caries in particular in their child population. METHODS: Assessments of the costs of treating existing and future caries by the conventional approach. Analysis of WHO dental databases and spreadsheet calculations of costs based upon population projections, prevalence and trends in patterns of caries. FINDINGS: Even though the caries levels are low and most of the disease occurs on the occlusal and the buccal/lingual surfaces, more than 90% of the dental caries remains untreated in Third World countries. Calculations reveal that to restore the permanent dentition of the child population of low-income nations using traditional amalgam restorative dentistry would cost between pounds 1,024 ($US1618) and pounds 2,224 ($US3513) per 1,000 children of mixed ages from 6 to 18 years. This exceeds the available resources for the provision of an essential public health care package for the children of 15 to 29 low-income countries. CONCLUSIONS: To treat caries with the traditional method of restorative dentistry is beyond the financial capabilities of the majority of low-income nations, as three-quarters of these countries do not even have sufficient resources to finance an essential package of health care services for their children. PMID- 11931217 TI - Comparative bond strengths of autopolymerising denture resin and light cured composite resin to denture teeth. AB - AIM: To investigate the shear bond strengths between light cured composite and autopolymerised acrylic resin bonded to acrylic resin denture teeth. METHODS: Surface treatments were used for the denture teeth included wetting with methylmethacrylate (MMA) monomer, composite bonding agent and acid etching. The samples were divided into seven groups. Two groups of specimens were immersed in distilled water for 30 days to hydrate the teeth. Five other groups of embedded denture teeth were stored in air at room temperature for 30 days. RESULTS: Analysis of variance indicated statistically significant differences between groups (P < 0.001). CONCLUSION: The greatest bond strengths to denture teeth were in acrylic resins polymerised with pressure and wetted with monomer. The bond strengths of hydrated and unhydrated samples gave similar results both with acrylic resins and composites. PMID- 11931218 TI - Dental attendance and anxiety among public and private school children in Jordan. AB - AIM: To evaluate the levels of dental attendance and anxiety among children of public (lower to middle socioeconomic groups) and private schools (higher socioeconomic groups), and to explore the perceived source of dental fear among them. METHOD: A group of 1,021 children selected by a simple random method from 10 public schools in the five geographic areas of Irbid Governate, Jordan and all 10 private schools were included in this study. All children (mean age 13.1 +/- 0.87 years) completed a questionnaire modified from the Dental Fear Scale (DFS) which also contained items related to dental attendance. RESULTS: Private school children (31.4%) visited the dentist regularly more significantly than public school children (15.0%) (P < 0.001). However, the majority of children (public = 82.6%, private = 67.4%) attended the dentist only in an emergency. The main reason for irregular attendance was 'treatment not needed' (42.2%). About 43-44% of children had dental fear of 'low to moderate type', while the prevalence of 'high dental anxiety' was slightly higher among children of public (11.6%) than those of private schools (6.9%). Fear of specific stimuli (pain and trauma) was the most common source of dental fear reactions among 60-65% of children. The sight and sensation of an anaesthetic needle and sight, sound and sensation of the drill were the most fear eliciting stimuli. Moreover, public school children were found to be more anxious with a significantly higher 'overall dental fear' than private school children (P < 0.05). PMID- 11931219 TI - Priorities in oral health care in non-EME countries. AB - This paper describes the prevailing problems pertaining to oral care in non established market economy (non-EME) countries. The current situation with large numbers of untreated cases of oral diseases, the inequality in delivery systems and the virtual non-existence of an adequate community oriented prevention calls for action. What is needed is a turn towards an oral care system that meets the principles of primary health care (PHC). This implies an oral health care system which makes use of the existing health care infrastructures and which applies an appropriate technology with emphasis on community oriented prevention directed to all at an affordable price. Four components of oral care are proposed as priorities in basic oral care, aiming to achieve the objectives of the PHC philosophy. These four components are: emergency care; exposure to fluoride; oral health education (OHE); atraumatic restorative treatment (ART). These components should be available for all. The exact content and extent of each component in various countries depends on local existing supporting conditions and on the level of development as well as on specific perceived needs of the population. Therefore, small-scale demonstration projects containing one or more of the described components should firstly be launched in various countries to evaluate the acceptability, effectiveness and sustainability of the proposed basic oral care programme. PMID- 11931220 TI - Periodontal Screening and Recording (PSR) Index: precursors, utility and limitations in a clinical setting. AB - Periodontal disease is a widespread condition ranging from simple gingival inflammation to severe periodontal breakdown. Early detection and diagnosis are key elements in the prevention of this oral pathology. Diagnostic aids available to dentists, such as the Periodontal Screening and Recording or PSR Index, accelerate the screening of periodontal conditions. While many studies provide support for the PSR Index as a valuable tool because it is reproducible, reliable, and quick, others have suggested that it may under or overestimate existing periodontal conditions. Some authors have suggested modifications to the recording method to improve its use either for screening purposes or for monitoring the outcome of periodontal therapy. This literature review briefly describes the PSR method, and compares studies based on both PSR and its counterpart, CPITN, in order to evaluate its usefulness and establish its limitations in a clinical setting. PMID- 11931221 TI - Restitution of maxillary anterior aesthetics with all-ceramic components. AB - This paper discusses the use of four types of ceramics; zirconia, leucite reinforced glass, densely-sintered pure aluminium oxide and a low fusing porcelain, for the restitution of anterior aesthetics. Recent advances in dental ceramic technology have made this scenario a reality, which only a decade ago, was thought elusive. While ceramics offer obvious advantageous properties of enhanced appearance, they also need to satisfy functional and longevity criteria to be considered viable restorative materials. Numerous studies have cited data to fulfil these criteria, including sustained function and survival rates similar to porcelain fused to metal (PFM) restorations. This discussion will focus on current dental ceramic advances, and present a clinical case study using all ceramic constituents to rectify a deteriorating dentition. PMID- 11931222 TI - Sex and the single chromosome. AB - Just as homology can trigger a chain of events as described in many of the chapters of this volume, sometimes a lack of homology causes a crisis of a different sort. So it is for the single X chromosome in XY males in many species. Divergent sex chromosome pairs, such as the X and Y chromosomes in mammals and in fruit flies, are thought to have evolved from homologous autosomes. During evolution, the Y chromosome has retained little coding capacity, leaving the male with reduced gene dosage for many functions encoded by the X chromosome. In this chapter we focus on dosage compensation in Drosophila, in which most X-linked genes are upregulated by a male-specific ribonucleoprotein complex. This complex is thought to recognize the X chromosome through approximately 35 dispersed chromatin entry sites and then spread in cis to dosage compensate most genes on the X chromosome. PMID- 11931223 TI - The origins of genomic imprinting in mammals. PMID- 11931224 TI - Genomic imprinting during seed development. AB - Genomic imprinting allows parent-of-origin specific control over gene expression. Although imprinted genes (or entire chromosomes) are homologous sequences that can be inherited from either parent, they are differentially marked by a heritable epigenetic modification (imprint), which can condition their behavior in term of gene expression. Imprinting-based regulation of entire chromosomes is observed in both insects (paternal genome elimination) and mammals (nonrandom X inactivation). Until recently, it was unknown whether plants possessed a similar epigenetic system discriminating between homologous chromosomes from either paternal or maternal origin. There is now experimental evidence for a genome-wide imprinting phenomenon during seed development in Arabidopsis. Genomic imprinting at the gene (or locus) level is observed in both mammals and flowering plants. In maize, only a few allelic variants of several nonessential genes expressed in the endosperm are imprinted. In Arabidopsis, gene-specific imprinting has recently been demonstrated for the MEDEA (and FIS2) gene, which is essential for normal seed development. Unlike the imprinted maize genes, so far all tested MEA alleles are subjected to regulation by imprinting. MEDEA and FIS 2 are members of the FIS class of genes (FERTILIZATION INDEPENDENT SEED) involved in regulation of growth and cell proliferation during seed development. MEDEA shares several paradigmatic features with imprinted mammalian genes. The MEDEA phenotypes provide empirical support for theories of an intragenomic parental conflict during seed development, whereby imprinting is proposed as a means to differentially balance the selfish interests of each sex's genome during the development of the progeny. PMID- 11931225 TI - Long-distance cis and trans interactions mediate paramutation. PMID- 11931227 TI - Is X-chromosome inactivation a homology effect? PMID- 11931226 TI - Homology-dependent gene silencing and host defense in plants. AB - Analyses of transgene silencing phenomena in plants and other organisms have revealed the existence of epigenetic silencing mechanisms that are based on recognition of nucleic acid sequence homology at either the DNA or RNA level. Common triggers of homology-dependent gene silencing include inverted DNA repeats and double-stranded RNA, a versatile silencing molecule that can induce both degradation of homologous RNA in the cytoplasm and methylation of homologous DNA sequences in the nucleus. Inverted repeats might be frequently associated with silencing because they can potentially interact in cis and in trans to trigger DNA methylation via homologous DNA pairing, or they can be transcribed to produce double-stranded RNA. Homology-dependent gene silencing mechanisms are ideally suited for countering natural parasitic sequences such as transposable elements and viruses, which are usually present in multiple copies and/or produce double stranded RNA during replication. These silencing mechanisms can thus be regarded as host defense strategies to foreign or invasive nucleic acids. The high content of transposable elements and, in some cases, endogenous viruses in many plant genomes suggests that host defenses do not always prevail over invasive sequences. During evolution, slightly faulty genome defense responses probably allowed transposable elements and viral sequences to accumulate gradually in host chromosomes and to invade host genes. Possible beneficial consequences of this "foreign" DNA buildup include the establishment of genome defense-derived epigenetic control mechanisms for regulating host gene expression and acquired hereditary immunity to some viruses. PMID- 11931228 TI - Quelling in Neurospora crassa. PMID- 11931229 TI - Non-Mendelian inheritance and homology-dependent effects in ciliates. AB - Ciliates are single-celled eukaryotes that harbor two kinds of nuclei. The germline micronuclei function only to perpetuate the genome during sexual reproduction; the macronuclei are polyploid, somatic nuclei that differentiate from the micronuclear lineage at each sexual generation. Macronuclear development involves extensive and reproducible rearrangements of the genome, including chromosome fragmentation and precise excision of numerous internal sequence elements. In Paramecium and Tetrahymena, homology-dependent maternal effects have been evidenced by transformation of the vegetative macronucleus with germline sequences containing internal eliminated sequences (short single-copy elements), which can result in a specific inhibition of the excision of the homologous elements during development of a new macronucleus in the sexual progeny of transformed clones. Furthermore, transformation of the Paramecium maternal macronucleus with cloned macronuclear sequences can specifically induce new fragmentation patterns or internal deletions in the zygotic macronucleus. These experiments show that the processing of many germline sequences in the developing macronucleus is sensitive to the presence and copy number of homologous sequences in the maternal macronucleus. The generality and sequence specificity of this transnuclear, epigenetic regulation of rearrangements suggest that it is mediated by pairing interactions between zygotic sequences and sequences originating from the maternal macronucleus, presumably RNA molecules. Alternative macronuclear versions of the genome can be maternally inherited across sexual generations, suggesting a molecular model for some of the long-known cases of non-Mendelian inheritance, and in particular for the developmental determination and maternal inheritance of mating types in Paramecium tetraurelia. PMID- 11931230 TI - RNAi (Nematodes: Caenorhabditis elegans). AB - RNA interference in Caenorhabditis elegans is a type of homology dependent posttranscriptional gene silencing induced by dsRNA. In this chapter we describe the history of the discovery of RNAi, its systemic nature, inheritance, and connection to other homology-dependent silencing phenomena like co-suppression and transcriptional gene silencing. We discuss RNAi-deficient mutants in C. elegans as well as characterized components of the RNAi, pathway, the molecular mechanism of RNAi, and its possible role in development and immunity. PMID- 11931231 TI - Antisense RNAs in bacteria and their genetic elements. AB - Antisense RNA-mediated regulation is widespread in bacteria. Most antisense RNA control systems have been found in plasmids, phages, and transposons. Fewer examples were identified in bacterial chromosomes. This chapter summarizes our current knowledge about antisense RNAs with respect to their occurrence, their biological roles, and their diverse mechanisms of action. Examples of cis- or trans-encoded antisense RNAs are discussed, and their properties compared. Most antisense RNAs are posttranscriptionally acting inhibitors of target genes, but a few examples of activator antisense RNAs are known. The implications of RNA structure on topologically and kinetically favored binding pathways are addressed, and solutions that have evolved to permit productive interactions between intricately folded RNAs are discussed. Finally, we describe how particular properties of individual antisense/target RNA systems match their respective biological roles. PMID- 11931232 TI - Transvection in Drosophila. AB - Pairing-dependent interallelic complementation was first described for the Ultrabithorax gene of the bithorax-complex in Drosophila by Lewis and cited as an example of a new phenomenon that Lewis called the "trans-vection effect." Several different kinds of pairing-dependent gene expression have been observed in Drosophila, and it is now clear that a variety of different molecular mechanisms probably underlie the changes in gene expression that are observed after disrupting chromosome pairing. Transvection in the bithorax-complex appears to result from the ability of cis-regulatory elements to regulate transcription of the promoter on the homologous chromosome. The same phenomenon appears to be responsible for pairing-dependent interallelic complementation at numerous other genes in Drosophila. Some transvection effects are dependent on the presence of wild-type or specific mutant forms of the protein encoded by the zeste trans regulatory gene, but other transvection effects are zeste-independent. The ease with which chromosome aberrations can disrupt transvection also varies widely among different genes. PMID- 11931233 TI - Pairing-sensitive silencing, polycomb group response elements, and transposon homing in Drosophila. AB - Regulatory DNA from a diverse group of Drosophila genes causes silencing of the linked reporter gene mini-white in the P-element vector CaSpeR. This silencing can occur in flies heterozygous for the P-element construct but is often enhanced in flies homozygous for the construct. In Drosophila, somatic chromosomes are paired and this pairing is important for the enhancement of silencing in most cases. Thus, this type of silencing has been called pairing-sensitive silencing. Many of the DNA fragments that cause pairing-sensitive silencing are regulatory elements required for the activity of the Polycomb group of transcriptional repressors (Polycomb group response elements, PREs). However, some PREs do not appear to cause pairing-sensitive silencing, and some fragments of DNA that cause pairing-sensitive silencing do not appear to act as PREs. I suggest that many PREs are composite elements of sites important for silencing and sites important for "pairing" or bringing together distant DNA elements. Both activities may be required for PRE function. In a related phenomenon, fragments of DNA included within P-element vectors can cause those transposons to insert in the genome near the parent gene of the included DNA (transposon homing). I suggest that DNA fragments that cause transposon homing or pairing-sensitive silencing are bound by protein complexes that can interact to bring together distant DNA fragments. PMID- 11931234 TI - Repeat-induced gene silencing in fungi. PMID- 11931235 TI - The evolution of gene duplicates. AB - Gene and genome duplications have given rise to enormous variability among species in the number of genes within their genomes. Gene copies have in turn played important roles in adaptation, having been implicated in the evolution of the immune response, insecticide resistance, efficient protein synthesis, and vertebrate body plans. In this chapter, we discuss the life history of gene duplications, from their first appearance within a population, through the period during which they rise in frequency or disappear, to their long-term fate. At each phase, we discuss the evolutionary processes that have influenced the dynamics of gene duplications and shaped their ultimate roles within a population. We argue that there is no evidence that organisms have evolved strategies to promote gene duplication in order to permit adaptive evolution. In contrast, many mechanisms exist to silence or eliminate duplicated genes, suggesting that selection has acted largely to reduce the rate of gene duplication. We also argue that natural selection has functioned as an effective sieve, increasing the representation of beneficial gene duplicates among those that establish within a population and that play a long-term role in evolution. To refine our understanding of how selection acts on new gene duplications, we provide a model incorporating a single-copy gene, its gene duplicate, and selection either favoring heterozygotes or eliminating deleterious mutations. Although both forms of selection can increase the initial rate of spread of a gene duplicate, the efficacy with which they do so differs dramatically. Heterozygote advantage always increases the rate of spread and can have a large impact. In contrast, masking deleterious mutations never has a large effect on the rate of spread of the duplicate, and this minor effect can be negative as well as positive. In both cases, the degree of linkage between the two gene copies affects the rate of spread of the duplication. Finally, we discuss evolutionary processes that occur over longer periods after a gene duplication has become established within a population. These long-term processes include maintenance, inactivation, and diversification in function. Consideration of each of the short-term and long-term processes affecting duplicated genes illustrates the subtle ways in which selection has acted to shape genomic structure. PMID- 11931236 TI - Prions of yeast as epigenetic phenomena: high protein "copy number" inducing protein "silencing". AB - Yeast infectious protein (prion) forms of the Ure2 and Sup35 proteins determine the nonchromosomal genes [URE3] and [PSI], and these are, therefore, the basis for a kind of epigenetic phenomena. In many systems, introduction of multiple copies of a DNA gene, or dsRNA copies of its sequence, results in the epigenetic silencing of that gene. In parallel with these homology effects, which act at the level of DNA or RNA, elevated copy number of the Ure2 and Sup35 proteins increases the frequency of their own "silencing" by prion formation. Both [URE3] and [PSI] appear to be due to self-propagating-amyloid formation of Ure2p and Sup35p, respectively. Another prion, [Het-s] of the filamentous fungus, Podospora anserina, is necessary for a normal cellular function, heterokaryon incompatibility. Since these prions are nonchromosomal genes, they are proteins acting as genes, a parallel to the fact that nucleic acids can catalyze enzymatic reactions. PMID- 11931237 TI - Homologous chromosome associations and nuclear order in meiotic and mitotically dividing cells of budding yeast. AB - This chapter discusses the relationship between nuclear order and the association of homologous DNA sequences in the budding yeast, Saccharomyces cerevisiae. Homologous chromosomes functionally interact with one another to repair DNA double-strand breaks (DSBs) introduced either environmentally (e.g., by gamma irradiation) or deliberately by the cell (e. g., during meiosis). DNA homology recognition in these instances often involves the (RecA) homolog RAD51 and/or the related gene, DMC1. Evidence for interactions between homologous chromosomes occurring independent of DSB formation and (RecA) homolog function has also been described in meiotic, premeiotic, and mitotically dividing cells of yeast. These interactions presumably depend upon DNA homology but the molecular details of such associations are poorly understood. Both DSB-dependent and -independent homolog associations may be facilitated by the nonrandom organization of chromosomes in the nucleus, including centromere and telomere clustering, which are also discussed. PMID- 11931238 TI - The role of sequence homology in the repair of DNA double-strand breaks in Drosophila. PMID- 11931239 TI - Effect of holding conditions on the detection of West Nile viral RNA by reverse transcriptase-polymerase chain reaction from mosquito (Diptera: Culicidae) pools. AB - We evaluated the effect of holding temperature and time between mosquito death and processing mosquito pools for virus detection on our ability to detect West Nile (WN) viral RNA from pools of mosquitoes by reverse transcriptase-polymerase chain reaction (RT-PCR). Pools of 24 uninfected Culex pipiens L. mosquitoes were "spiked" with either a single Cx. pipiens that had been inoculated previously with WN virus or with an uninfected mosquito. These pools were held dry at 20, 4, -20, or -70 degrees C for selected time intervals before all mosquito pools were triturated in TRIzol LS reagent and processed for detection of WN viral RNA. While infectious virus virtually disappeared from pools maintained at 20 degrees C by 48 h after mosquito death, neither holding temperature (20 to -70 degrees C) nor holding period (up to 2 wk) affected detection of WN viral RNA by real-time RT-PCR. These findings suggest that we need not keep mosquitoes chilled to be able to detect WN viral RNA effectively by RT-PCR. This should enhance the feasibility of field-based WN virus surveillance programs where only detection of WN viral RNA is the objective and maintenance of a cold chain may not be possible. PMID- 11931240 TI - Seasonal variation in Carcinops pumilio (Coleoptera: Histeridae) dispersal and potential for suppression of dispersal behavior. AB - Seasonal dispersal of Carcinops pumilio (Erichson) was evaluated using two trapping methods-a black-light pitfall trap and a mesh-bottomed trap placed on poultry manure. The black-light trap collected larger numbers than the mesh bottomed trap from March through June. The mesh-bottomed trap gathered larger numbers of beetles from June through August and numbers were less variable throughout the year. Often, when very low numbers of beetles were recovered from manure cores, large numbers of beetles could be collected with the black-light trap suggesting that beetle density may not be an important factor in dispersal behavior. The greatest dispersal in the dispersal arenas (approximately 90%) occurred using beetles collected by both trap types in June 2000. Beginning in March and ending in August, a cyclic rise and then fall pattern in both laboratory dispersal and beetle collections was observed. Trap collection patterns were similar in both years of the study. In January and March, we were unable to prevent dispersal behavior of beetles captured in black-light traps. However, in May, after beetles had been in a dispersal phase for several months, we were able to suppress dispersal. In contrast, dispersal behavior among beetles captured with the mesh-bottomed trap did not change following the photoperiod altered exposure. PMID- 11931241 TI - Ticks parasitizing dogs in northwestern Georgia. AB - From January 1998 through September 1999, 324 dogs in three northwestern Georgia counties were examined for ticks. Six species of ticks were recovered. The three most commonly collected ticks were the American dog tick, Dermacentor variabilis (Say) (310 male male, 352 female female; prevalence, 97%; mean intensity 2.1); the brown dog tick, Rhipicephalus sanguineus (Latreille) (118 male male, 119 female female, 38 nymphs; prevalence, 22%; mean intensity, 3.8); and the lone star tick, Amblyomma americanum (L.) (8 male male 26 female female, 2 nymphs; prevalence, 5%; mean intensity, 2.4). Other ticks recovered were Ixodes cookei Packard (3 female female); the Gulf Coast tick, Amblyomma maculatum Koch (2 female female); and the blacklegged tick, Ixodes scapularis Say (1 female). Another adult female specimen of I scapularis was recovered from a cat, further reinforcing that this medically important tick is present in northwestern Georgia. PMID- 11931242 TI - Mosquito abundance is correlated with cliff swallow (Petrochelidon pyrrhonota) colony size. AB - We measured the abundance of mosquitoes [primarily Aedes vexans (Meigen) and Culex tarsalis Coquillett] at cliff swallow (Petrochelidon pyrrhonota Vieillot) colonies of different sizes in southwestern Nebraska in 1999. Using CO2 traps placed inside and outside of colonies, we found that total mosquito abundance increased significantly with the number of active cliff swallow nests at a colony site. We found no effect of date or weather conditions on the number of mosquitoes caught it the different sites. By classifying the landscape from aerial photographs within a 2-km-diameter circle centered on each colony site, we found no significant relationships between habitat type near a colony site and cliff swallow colony size or mosquito abundance. Proximity to livestock could not account for our results. Culex tarsalis was proportionately more likely to be caught inside a colony than at traps 30 in away, but the proportion of C. tarsalis inside a colony did not vary with colony size. Our results cannot be explained by date- or weather-related sampling artifacts or by differences in habitat between sites. Most likely, mosquitoes, especially A. vexans, are attracted to the vicinity of large cliff swallow colonies. PMID- 11931243 TI - Sand flies (Diptera: Phlebotominae) in Sanliurfa, Turkey: relationship of Phlebotomus sergenti with the epidemic of anthroponotic cutaneous leishmaniasis. AB - Sand fly (Diptera: Phlebotominae) fauna were surveyed in various districts of Sanliurfa in southeast Turkey for 3 yr immediately after an epidemic of cutaneous leishmaniasis (Leishmania tropica). Sticky papers and CDC light traps collected a total of 10,937 sand flies, of which 10,919 (4,158 females and 6,761 males) were identified as Phlebotomus and 18 (11 females and seven males) as Sergentomyia (S. theodori Parrot; S. adleri Theodor). Eight Phlebotomus spp. were identified: P. sergenti Parrot (72.3%), P. papatasi (Scopoli) (27.2%), P. brevis Theodor & Mesghali (0.20%), P. neglectus Leger & Pesson (0.13%), P. perfiliewi Parrot (0.05%), P. mascitti Grassi, P. halepensisTheodor, and P. alexandri Sinton (0.01%). Phlebotomus mascitti and P. neglectus, along with both Sergentomyia sp., have not been previously described from the study area. Similar results were obtained when both trapping methods were applied in the same houses, indicating that local P. sergenti and P. papatasi populations were equally attracted to the light. P. sergenti was consistently abundant, agreeing with the general view that this species is the vector of leishmaniasis in the region. There was no apparent decrease in the relative abundance of this vector versus the other species, suggesting that factor (s) other than a change in the dynamics of sand fly populations precipitated the decline of the human leishmaniasis epidemic in Sanliurfa. PMID- 11931244 TI - Olfactory cues for oviposition behavior in Toxorhynchites moctezuma and Toxorhynchites amboinensis (Diptera: Culicidae). AB - In laboratory tests gravid female Toxorhynchites moctezuma (Dyar & Knab) and Toxorhynchites amboinensis (Doleschall) were offered the choice of black oviposition jars containing a diethyl ether extract of water collected from a natural oviposition site for these species (i.e., used tires), a dilution series of 4-methylcyclohexanol, 3-methylindole, 2-methylphenol, 3-methylphenol and 4 methylphenol, or solvent. Tire water extract and all test compounds acted as oviposition attractants and stimulants for both species, with the threshold amounts required to elicit these behaviors varying between the species and among the compounds tested. PMID- 11931245 TI - Attractiveness of beach ball decoys to adult Stomoxys calcitrans (Diptera: Muscidae). AB - The attractiveness of inflated beach balls covered with adhesive and used as decoys to trap adult stable flies was investigated on Florida panhandle beaches. Decoys were painted either solid black, solid white, or a mixed pattern that consisted of three equally spaced white circles (20 cm diameter) on a solid black background. Another set of decoys (referred to as plain) were unpainted and retained the manufacturer's original color scheme. The plain decoy consisted of a separate blue, yellow, and red diamond-shaped panel. Each color panel was separated by a white panel of similar size and orientation. Plain decoys collected significantly (<0.05) more stable flies than other treatments but no significant difference was noted between colored panels. The mixed pattern decoy captured significantly fewer flies than the plain decoy but significantly more flies (nearly twice) when compared with solid white or black decoys. No difference in preference was observed when fly abundance on the black background was compared with that on white circles and total abundance from both areas appeared to be additive compared with either area alone. No significant differences were found in the number of flies trapped on solid white versus black decoys. When trapping efficiency was compared with Alsynite translucent fiberglass cylinders covered with adhesive-treated cellophane sheets, the decoy trap caught significantly more (>10 times) flies per square centimeter. Alsynite cylinders are considered standard tools when sampling fly populations. Adhesive treated beach ball decoys may be an alternative method for luring stable flies away from human hosts in recreation areas, or from animals, thereby reducing biting annoyance from these pests. PMID- 11931246 TI - Responses of fertile and sterile screwworm (Diptera: Calliphoridae) flies to bovine blood inoculated with bacteria originating from screwworm-infested animal wounds. AB - ABSTRACT A simple bioassay system was developed to study locomotory and ovipositional responses of screwworm, Cochliomyia hominivorax (Coquerel), flies to bovine blood inoculated with eight species of coliform bacteria that were isolated from screwworm-infested animal wounds. When exposed to odors from bacteria-inoculated blood which was incubated for 72 h at 37 degrees C, approximately 50% of 7- and 10-d-old gravid females landed on the blood by the end of 15 min test exposure. Only 17% of 7-d-old reproductively sterile females (from irradiated pupae) with previtellogenic ovaries and 2% of 4-d-old vitellogenic females responded to the same treatment. Females generally reacted in greatest numbers to bacteria-inoculated blood incubated for 72 h, followed by 48 h, then 24 and 96 h. Males of all ages tested were unresponsive. Although oviposition occurred in tests with gravid females lasting for 1 h, with both inoculated blood and an uninoculated control, the inoculated sample was significantly better than the control at 48, 72, and 96 h incubation duration. Our results are consistent with the conclusion that the inoculated blood, when incubated for 48-72 h, gives off volatile chemicals which attract gravid females and contains an oviposition stimulant that acts following contact and feeding. The volatiles, once isolated and identified, may be useful for sampling gravid females in the field as well as improving the oviposition system in the mass production facility of the screwworm eradication program. PMID- 11931248 TI - Epidemiological significanceof subterranean Aedes aegypti (Diptera: Culicidae) breeding sites to dengue virus infection in Charters Towers, 1993. AB - The objective of this study wasto determine the epidemiological significance of subterranean mosquito breeding sites to the 1993 outbreak of dengue fever (type 2) in the northern Queensland town of Charters Towers, Australia. In recent studies on subterranean mosquito breeding, containers such as wells and service manholes have been shown to be important breeding sites to Australia's only dengue vector, Aedes aegypti (L.). This study demonstrates a direct epidemiological association between subterranean breeding sites and dengue virus infection. The mean distance between residents seropositive for dengue 2 and the nearest subterranean container (113 m) was significantly less than for a randomly selected control (191 m), (F = 81.9; df = 1, 478; P < 0.001). Residents positive for dengue 2 antibodies was 2.47 (95% confidence interval 1.88-3.24) times higher for those living within 160 m of a well or service manhole, compared with those residing further away. These findings emphasize the importance of incluuding subterranean water containers in Ae. aegypti surveillance and control programs. PMID- 11931247 TI - Taxonomic status of Ixodes didelphidis (Acari: Ixodidae). AB - Ixodes didelphilis Fonseca & Aragao was described in Brazil in 1952 as a new tick species that differed from Ixodes loricatus Neumann by the spiracular plate pattern. We have reared four tick colonies from different geographic areas in the laboratory that were started from single engorged females originally identified as I. didelphidis (BMG, colony ) and I. Iocicatus (CSP, PSP, and TRJ colonies). We analyzed the spiracular plate morphology of F1 adult ticks from each tick colony, compared their biological 11th, and performed a molecular analysis of the second internal transcribed rDNA spacer (ITS2) to test the validity of the species I. didelphidis. The spiracular plate analysis of laboratory F1 adult ticks yielded single females from the four colonies showed variations that invalidate morphological parameters for differentiation of loricatus and I. didelphidis. Biological data of the BMC, CSP, and TRJ colonies were similar. The biology ofthe PSP colony was not evaluated. The ITS2 sequence variations observed between the tick colonies ranged from 1.3 to 4.9%, and the similarity tree constructed by the neighbor-joining method with nucleotide distances showerl that the distances between the samples were similar to what is expected for intraspecific variations found in other ticks species. The morphological and biological results, in conjunction with the ITS2 analysis, supported the conspecificity I. loricatus and I. didelphidis. PMID- 11931249 TI - Susceptibility of the mosquitoes Anopheles minimus, An. sinensis, and An. saperoi (Diptera: Culicidae) from the Ryukyu Archipelago, Japan, to the rodent malaria Plasmodium yoelii nigeriense. AB - The susceptibility of three anopheline mosquitoes, Anopheles minimus Theobald, An. sinensis Wiedemann, and An. saperoi Bohart & Ingram, from the Ryukyu Archipelago to the rodent malaria, Plasmodium yoelii nigeriense was examined to find new vectors other than An. stephensi Liston for rodent malaria studies in the laboratory. The survival rate of the mosquitoes after feeding on mice infected with P. y. nigeriense was also examined. The Beech strain of An. stephensi from India was compared with An. minimus from Ishigaki Island, and An. sinensis and An. saperoi from Okinawa Island. Oocysts were first found on day 3 after feeding on mice infected with P. y. nigeriense in An. stephensi, on day 4 in An. minimus and An. saperoi, and day 6 in An. sinensis. From 8 to 14 d after feeding on malaria-positive mice, oocysts were present in 97.2-100% of An. stephensi, 85.7-100% of An. saperoi, 20-74.1% of An. minimus, and 12.5-13.3% of An. sinensis. The duration of oocyst occurrence in An. saperoi was 55 d, the longest among the anopheline mosquitoes used in this study. On day 8 after feeding, sporozoites were found in the salivary glands and heads of all the mosquitoes tested. From the 10th to 16th d, sporozoites were present in the salivary glands of 14.9% (range, 9.1-28.0%) of An. minimus, 47.3% (40.7-58.1) of An. saperoi, and 96.2% (94.1-97.2) of An. stephensi, but were absent in An. sinensis. Anopheles saperoi could be an excellent vector of P. y. nigeriense because it has comparatively high susceptibility and high longevity even after feeding on infected mice. PMID- 11931250 TI - Laboratory evaluation of fipronil and imidacloprid topical insecticides for control of the plague vector Oropsylla montana (Siphonaptera: Ceratophyllidae) on california ground squirrels (Rodentia: Sciuridae). AB - Two insecticides, fipronil and imidacloprid, were evaluated for efficacy and longevity against Oropsylla montana (Baker), the most important vector of plague in California. Wild-caught California ground squirrels, Spermophilus beecheyi (Richardson), were individually housed in the laboratory to serve as natural hosts to O. montana and for on-animal insecticide trials. Several concentrations oftechnical grade fipronil and imidacloprid in acetone were applied to samples of clean rodent bedding to determine residual activity and longevity against fleas. Immature and adult cat fleas, Ctenocephalides felis (Bouche), were used as representative fleas for periodic assays in place of less fecund O. montana. Toxicity of treated bedding did not decrease significantly for 1 yr at all applied concentrations. Fipronil provided 100% kill for at least 1 yr at > or = 100 ppm, whereas imidacloprid required 10,000 ppm for similar performance. Laboratory squirrels were treated with topical formulations of fipronil (Frontline Top Spot) and imidacloprid (Advantage Flea Adulticide) at a dosage rate of 15 mg/kg and evaluated for residual activity every 2 wk against adult O. montana. Residual activity was determined by percent recovery of O. montana adults released on treated and untreated animals after 48 h. Frontline provided 100% kill of adult fleas for at least 10 wk, and up to 26 wk on one animal. Advantage failed to provide 100% kill of adult fleas at 2 wk, with complete loss of efficacy by week 6. Concurrent assays with bedding samples from squirrel nest boxes showed negligible toxicity transfer from treated animals to nest bedding. PMID- 11931251 TI - Speciation and distribution of the members of the Anopheles punctulatus (Diptera: Culicidae) group in Papua New Guinea. AB - Mosquito collections were made throughout the mainland of Papua New Guinea to identify the members of the Anopheles punctulatus group present and to determine their distribution. Identification was made using morphology, DNA hybridization, and polymerase chain reaction (PCR)-RFLP analysis. Nine members of the group were identified: An. farauti s.s. Laveran, An. farauti 2, An. koliensis Owen, and An. punctulatus Donitz, were common and widespread; An. farauti 4 was restricted to the north of the central ranges where it was common; An. farauti 6 was found only in the highlands above 1,000 m; and An. farauti 3, An. sp. near punctulatus and An. clowi Rozeboom & Knight were uncommon and had restricted distributions. Identification of An. koliensis and An. punctulatus using proboscis morphology was found to be unreliable wherever An. farauti 4 occurred. The distribution and dispersal of the members of the An. punctulatus group is discussed in regard to climate, larval habitats, distance from the coast, elevation, and proximity to human habitation. PMID- 11931252 TI - Density-dependent development of Anopheles gambiae (Diptera: Culicidae) larvae in artificial habitats. AB - The growth and development of Anopheles gambiae Giles larvae were studied in artificial habitats in western Kenya. Larvae responded to increasing densities by extending their development time and by emerging as smaller adults, although survival was not significantly affected. Addition of nutrients in the form of cow dung collected near the study site had no impact on larval growth and development. Regression analysis showed that female development time increased by 0.020 d and female dry mass decreased by 0.74 microg with each additional larva. By fitting the data to the pupation window model, the estimated minimum dry mass to achieve pupation was 0.130 mg and the estimated minimum time to pupation was 5 d. The most likely food source for An. gambiae larvae was algal growth, which was significantly reduced by the presence of larvae. Bacterial densities were not significantly affected by the presence of larvae although total bacteria counts were lower at the higher densities indicating they may provide a secondary food source when algal resources are depleted. Similarly, the levels of nitrogen and phosphorus in the habitats were not significantly affected by the presence of larvae although there was evidence of decreasing nitrogen levels occurring with increasing larval densities suggesting that nitrogen may be a limiting resource in the larval environment. The data indicate that competition within the larval environment may indirectly regulate An. gambiae populations by reducing adult body size, which may in turn reduce adult survivorship and fecundity. The potential impact of density-dependent interactions among An. gambiae larvae on the transmission of Plasmodium falciparum is discussed. PMID- 11931253 TI - Immune responses of interferon gamma (IFN-gamma) knock out mice to repeated Haemaphysalis longicornis (Acari: Ixodidae) nymph infestations. AB - To investigate the immunological mechanisms of acquired resistance to tick infestation, interferon gamma (IFN-gamma) deficient mice (IFN-gamma mice) were used to assess interleukin-4 (IL-4) and antibody production levels against tick salivary gland antigen on three successive infestations with Haemoaphysalis longicornis Neumann nymphs. The engorged body weight of the ticks decreased during the second and third infestations. Similar observations were noted in IFN gamma+/+ mice. However, the engorged body weight of the ticks from IFN-gamma +/+ mice were considerably lower than those from IFN-gamma-/- mice. A marked increase in antibody production during the second and third infestations was observed indicating that IFN-gamma-/- mice could acquire immunological resistance against H. longicornis nymphs. Moreover, IL-4 levels were higher during the first and third infestations but decreased during the second infestation. IL-4 levels were significantly higher in IFN-gamma-/- mice than in IFN-gamma+/+ mice. We have shown here that the statistically significant high IL-4 levels observed in IFN gamma-/- mice may be a result of type 2 helper cell (Th2) polarization. However, the apparently higher IL-4 levels during the first and third infestations and the notable decline during the second infestation suggest that other cytokines or factors in the host immune system may play a part in regulating IL-4 levels. PMID- 11931254 TI - Distribution of Borrelia burgdorferi sensu lato in Ixodes ricinus (Acari: Ixodidae) ticks from the Basque Country, Spain. AB - Borrelia burgdorferi was found widespread in ixodid ticks from the Basque Country (Spain) during a two-step study. In the first part, a total of 7,835 ixodids of eight different species was collected from vegetation, classified, and processed using polymerase chain reaction (PCR) for detection of B. burgdorferi ospA DNA. B. burgdorferi DNA wasdetectedin < or = 12.5% of adults and > or = 0.6% of Ixodes ricinus (L., 1758) nymphs (mean 1.5 and 0.05%, respectively), and in < or = 14.3% of adult Hemaphysalis punctata (Canestrini & Fanzago, 1877) analyzed (mean 1.2%). The second part of the study was undertaken 2 yr later to characterize B. burgdorferi distribution by focusing on the areas where L. ricinus was the predominant species. Ten areas were selected from which 1,535 nymphs and adults of I. ricinus were collected and processed by PCR and culture techniques. Infected ticks were found in all zones. B. burgdorferi DNA was detected in a mean of 9.3 and 1.5% of adults and nymphs, respectively. Nine isolates of B. burgdorferi were obtained, belonging to four different genospecies (B. burgdorferi sensu stricto, B. garinii, B. valaisiana, and B. lusitaniae). The results indicate that some areas of Spain have a potential risk for Lyme disease agent exposure and that B. borgdorferi appears to have an increasing occurrence in ticks in the Basque Country. PMID- 11931255 TI - Reproductive activity and survival of Culex pipiens pallens and Culex quinquefasciatus (Diptera: Culicidae) in Japan at high temperature. AB - The egg hatchability, insemination, and longevity of Japanese Culex pipiens pallens Coquillett and Japanese Culex quinquefasciatus Say were compared at 25 and 30 degrees C. Egg hatchability was high in Cx. p. pallens at 25 degrees C, but it was very low at 30 degrees C because almost no females were inseminated at this temperature. In Cx. quinquefasciatus, the egg hatchability and insemination rates were very high, even at 30 degrees C. The longevity of adult females and males was generally shorter in Cx. p. pallens than in Cx. quinquefasciatus at both temperatures. Because high temperatures may restrict the spread of Cx. p. pallens, we suggest that even if this species spreads to Okinawa, the possibility of it becoming established is very low. PMID- 11931256 TI - Attractant volatiles released by female and male Triatoma infestans (Hemiptera: Reduviidae), a vector of chagas disease: chemical analysis and behavioral bioassay. AB - Volatiles emitted by male and female T infestans before and during copula were collected on Porapak-Q filters, desorbed with dichloromethane, and analyzed by gas chromotography and gas chromatography-mass spectrometry after confirmation of attractiveness in an arena bioassay. Chemical analysis confirmed the presence of (R,S) -2- and 3-methylbutan-1-ol in a 2:1 ratio; short chain acids (ethanoic to nonanoic acid); long chains acids decanoic to (Z)-9-octadecenoic acid; aliphatic aldehydes (hexanal to nonanal), benzaldehyde and dipropylsulphide from insects in copula. Electroantennographic studies conducted with a homologous series of aliphatic aldehydes on female and male T infestans showed that, for a given dose, EAG responses elicited from both sexes increased with increased chain length up to nonanal, after which EAG-activity declined. Attractiveness of non-acidic trace components identified in the volatiles were tested on male and female T. infestans, in an arena bioassay using a video tracking method. Aliphatic C6 to C10 aldehydes were tested: hexanal (1-100 microg) and heptanal (10 microg) were attractive to female T. infestans, high doses of octanal and nonanal (1-100 microg) were Unattractive to male and female T. infestans but low doses of nonanal (0.01-0.1 microg) were attractive to male T infestans. Benzaldehyde was highly attractive to female T. infestans at low doses (0.05- 0.1 microg). 3 methylbutan-1-ol was attractive to male T infestans at high dose (1,000 microg). (S) or (S,R) 2-methyl-butan-1-ol were attractive to males or females (1-1,000 microg). Blends of hexanal and benzaldehyde (20:1 and 40:1) showed an additive effect on attraction compared with hexanal alone, when tested on female T. infestans. The study has demonstrated the presence of a number of electrophysiologically and behaviorally active compounds in volatiles emitted by T. infestans in copula that may have a role in the postulated copulation pheromone. PMID- 11931257 TI - Prevalence of borrelia burgdorferi sensu lato infection among rodents and host seeking ticks in South Carolina. AB - Tissues of rodents and host-seeking adult ticks collected in the Piedmont, Sandhills, Coastal Plain, and Coastal Zone of South Carolina were cultured in attempts to isolate Borrelia burgdorferi (Johnson, Schmid, Hyde, Steigerwalt & Brenner), the etiologic agent of Lyme disease. An exploratory, tree-based statistical analysis was used to identify ecological variables that were associated with spirochete infection among rodents and ticks. Spirochetes were isolated from tissues of 71 rodents: 22 (69%) of 32 eastern woodrats, 39 (53%) of 74 cotton mice, and 11(25%) of 44 hispid cotton rats. Rodent infection prevalences were significantly higher in the Coastal Zone than in other regions. Spirochetes were also cultured from 31 (2.6%) of 1,193 questing ticks. Prevalence of spirochetes in Ixodes affinis Neumann (19/74, 26%) was significantly higher than in I. scapularis Say (12/864, 1.3%) and other species (0/255) of ticks tested. In addition, two (9%) of 23 adult I. minor Neumann removed from woodrats contained spirochetes. Isolates from rodents and ticks were analyzed immunologically by indirect immunofluorescence and Western blots, and further characterized by polymerase chain reaction assays and sodium dodecyl sulfate polyacrylamide gel electrophoresis. All were determined to be B. burgdorferi sensu lato. Results of this study confirmed that B. burgdorferi is endemic in South Carolina, and that enzootic transmission cycles exist at foci in the Coastal Zone. These findings add additional evidence that I. affinis and I. minor are potentially significant maintenance vectors of the spirochete. PMID- 11931258 TI - Induced immunity against the mosquito Anopheles stephensi (Diptera: Culicidae): effects of cell fraction antigens on survival, fecundity, and plasmodium berghei (Eucoccidiida: Plasmodiidae) transmission. AB - Two subeellular fractions from the midgut of the malaria mosquito Anopheles stephensi (Liston) were used to immunize BALB/c mice. Mice were subsequently infected with the rodent malaria parasite Plasmodium berghei (Vineke & Lips), and the effects of anti-mosquito immunity on mosquito survival and fecundity and on parasite transmission were investigated. Mosquitoes were infected directly from mice (in vivo) or by feeding cultured ookinetes through a membrane (in vitro). Infections were monitored by counting oocysts on the midgut wall. Microvilli extracts induced a strong and partially specific antibody reaction against the midgut, which was manifest as decreased survival in in vivo fed mosquitoes and reduced fecundity in both kinds of feeding. Antisera against microvilli reduced the mean intensity of P. berghei oocysts when fed in vitro, while mosquitoes fed antiserum against basolateral plasma membranes in vivo, showed higher oocyst burdens. PMID- 11931259 TI - Susceptibility of mansonia indiana (Diptera: Culicidae) to nocturnally subperiodic Brugia malayi (Spirurida: Filariodea). AB - Mosquitoes, Mansonia indiana Edwards, 1930, were collected from non-endemic area of human lymphatic filariasis and tested for their susceptibility of infection using nocturnally subperiodic Brugia malayai Buckley & Edeson, 1956. Three cats naturally infected with B. malayi were used in the experiment for mosquitoes feeding. The data revealed that the susceptibility of mosquito infection ranged from 30 to 70%. The results also revealed that the susceptibility rates were not linearly correlated to the microfilarial densities in the cat at the time of feeding. The microfilarial density in cats ranged from 15 to 27 per 10 microl of blood whereas the mean number of third stage larvae in the infective mosqiitoes ranged from 21.6 to 26.8. In addition, statistical analysis showed no significant difference (P > 0.05) between the mean number of third-stage larvae in mosquitoes and the density of microfilaria in cats. The study indicated that Ma. indiana, collected from non-endemic areas, is capable for transmitting the nocturnally subperiodic B. malayi. PMID- 11931260 TI - The prevalence of Borrelia burgdorfieri (Spirochaetales: spirochaetaceae) and the agent of human granulocytic ehrlichiosis (Rickettsiaceae: Ehrlichieae) in Ixodes scapularis (Acari:Ixodidae) collected during 1998 and 1999 from Minnesota. AB - We tested 103 adult Ixodes scapularis Say from 12 counties in Minnesota for the presence of Borrelia burgdorferi and the causative agent of human granulocytic ehrlichiosis (HGE), using polymerase chain reaction (PCR). A total of 17 ticks (16.5%) was positive for B. burgdoiferi using nested PCR for the flagellin gene. or both PCR for the ospA gene and nested PCR for the flagellin gene. A total of four ticks (3.8%) was positive for the agent of HGE using nested PCR for 16S rDNA. Counties in Minnesota with established and recently reported populations of I. scapularis both had ticks infected with B. burgdorferi. The agent of HGE was only detected in counties with established I. scapularis populations. PMID- 11931261 TI - Effect of environmental temperature on the ability of Culex pipiens (Diptera: Culicidae) to transmit West Nile virus. AB - Environmental temperature can affect the ability of mosquitoes to transmit an arbovirus. However, results of various studies indicate that these effects are not consistent among viruses or mosquito species, and there is no information available on the effect of environmental temperature on the ability of North American mosquito species to transmit West Nile (WN) virus. We evaluated the effect of incubation temperature (18, 20, 26, or 30 degrees C) on the ability of Culex pipiens L. derived from specimens collected during the outbreak in New York in 1999 to transmit a strain of WN virus obtained from a crow that died during this outbreak. Although mosquitoes fed on the same viremic chickens, infection rates were directly related to subsequent incubation temperatures. In mosquitoes held at 30 degrees C, virus was recovered from nearly all mosquitoes tested, disseminated infections were detected as early as 4 d after the infectious blood meal, and >90% of all mosquitoes had a disseminated infection 12 or more days after the infectious blood meal. In contrast, for mosquitoes held at 18 degrees C, disseminated infections were not detected until 25 d after the infectious blood meal, and even after 28 d, <30% contained a disseminated infection. Results for mosquitoes held at 20 and 26 degrees C were intermediate for both infection and dissemination rates. The effect of environmental temperature should to be considered when evaluating the vector competence of these mosquitoes and modeling risk of WN virus transmission in nature. PMID- 11931262 TI - Simulated overwintering of encephalitis viruses in diapausing female Culex tarsalis (Diptera: Culicidae). AB - Female Culex tarsalis Coquillett in reproductive diapause were infected per os or by intrathoracic inoculation with western equine encephalomyelitis (WEE) or St. Louis encephalitis (SLE) viruses during "fall," maintained over a simulated "winter," and then tested for virus infection and transmission in vitro and in vivo after "vernal" termination. Exposure of F1 progeny of field-collected females to cool temperatures and short daylength produced females in reproductive diapause that were reluctant to imbibe infectious virus from pledgets soaked with suspensions of virus, blood and sucrose (2.5% by volume). Those infected per os maintained virus at very low or undetectable titers. Some females that originally tested negative for WEE by plaque assay on Vero cell culture tested positive by reverse transcriptase-polymerase chain reaction (RT-PCR) and by Vero cell culture after passage in mosquito cells. Few females became infected orally with SLE, but these infected females developed elevated titers. Females inoculated with SLE retained their infection through winter and then transmitted readily in vitro and in vivo. Feeding on a vertebrate host after diapause termination significantly increased the titer of SLE in previously infected females. These experiments simulated how infections acquired either horizontally or vertically may provide mechanisms for WEE and SLE overwintering. Attempts to detect infected females during winter following a summer with enzootic WEE activity were negative by both RT-PCR and plaque assay. PMID- 11931263 TI - Comparative behavior of different life-cycle stages of Ixodes ricinus (Acari: Ixodidae) to human-produced stimuli. AB - Nymphs of Ixodes ricinus (Linnaeus, 1758), I. scapularis Say, 1821, and I. pacificus Cooley & Kohls, 1943 are epidemiologically the most dangerous stage for transmission of Lyme disease to humans. Many factors play a role in the epidemiological significance of the nymphs. In this study, we address the question of whether nymphs show a greater tendency than adults to accept humans as their host. To evaluate this, we have compared the host acceptance behavior of nymphs and adults (males and females) with respect to a human in Rambouillet forest, a focus of Lyme disease. Individual ticks (nymph, male or female) located on a herbaceous stem were exposed to different stimuli (e.g., approach, stem movement, breathing), and the response of each individual to these stimuli was noted. Tick responses were categorised into classes (from 0 to 3) according to their intensity. Statistical analysis carried out on 22 ticks allowed us to compare the behavior of the nymph stage with respect to males and females. Despite the small sample sizes, it appears that nymphs are more responsive to a human than are the adults. PMID- 11931264 TI - A survey for nymphs of host-seeking Ixodes scapularis and Amblyomma americanum (Acari: Ixodidae) on tree trunks in deciduous forests. AB - Trunks of 83 trees in a mixed deciduous forests in Maryland were sampled for the presence of nymphs of the blacklegged tick, Ixodes scapularis Say, and the lone star tick, Amblyomma americanum (L.). Although one or more nymphs of either I. scapularis or A. americanum was found in leaf litter and substrate < or = 1 m from the bases of 47% of the trees sampled, a total of 6 I. scapularis nymphs was found on the trunks of only five trees. No nymphs were found on the trunks of 12 dead trees. No A. americanium nymphs were found on any tree trunks. The trunks were sampled to 2.5 m above the soil, but the nymphs were found < or = 1 m from the ground. More than 50% of I. scapularis nymphs found in the leaf litter < or = 1 m from bases of living trees were north of the trees sampled, whereas few I. scapularis were found west of trees. These findings suggest that the I. scapularis nymphs' presence on tree trunks is of little ecological consequence, unless nymphs were being removed from tree trunks by acquiring hosts at such a rapid rate that nymphal numbers on trunks could not accrue. PMID- 11931265 TI - Parasitism of domestic swine (Sus scrofa) by amblyomma ticks (Acari: Ixodidae) on a farm at Monte Negro, western Amazon, Brazil. AB - ABSTRACT In January 2001, while conducting a survey of the tick fauna of the State of Rondjnia, Brazil, a rural area within Monte Negro county was visited. On one farm within the county the producer maintained a herd of crossbred swine, Sus scrofa L., that was reared under unconfined conditions, with unrestrained access to the pasture and adjacent native Amazon equatorial forest. Inspection of the swine herd produced a total of 77 ticks collected from eight adult pigs (mean, 9.6 ticks per pig) that were identified as Amblyomma naponense (Packard) (26 males, eight females), A. oblongoguttatum Koch (five males, three females), A. ovale Koch (one female) and A. scalpturatum Neumann (one male). One Amblyomma larva and 32 Amblyomma nymphs also were collected from the pigs. Of these, six nymphs were reared in the laboratory until they reached the adult stage, one being an A. oblongoguttatum female and five being A. scalpturatum females. PMID- 11931266 TI - Evaluation of anopheline mosquitoes (Diptera: Culicidae) from the republic of Korea for Plasmodium vivax circumsporozoite protein. AB - As part of an on-going malaria surveillance effort conducted by the U.S. Forces Korea, Republic of Korea (ROK), a total of 28,286 anopheline mosquitoes was tested for the presence of Plasmodium vivax circumsporozoite (CS) protein. Mosquitoes were collected (using a variety of light and baited traps) from 29 locations throughout the ROK (the majority were collected near the de-militarized zone), identified to species, and tested by enzyme-linked immunosorbent assay for the presence of P. vivax 210 and P. vivax 247 CS protein. Recent evidence suggests that characters used to separate Anopheles sinensis Wiedemann from An. lesteri Baisas & Hu are unreliable; therefore, the data have been analyzed by grouping these two species. A total of 25,365 Anopheles sinensis/lesteri, 2,890 An. yatsushiroensis Miyazaki, and 31 An. sineroides Yamada was tested. Of these, one pool of 10 An. sinensis/lesteri collected on 9 September 1999 at Camp Howze and one pool of nine An. sinensis/ lesteri collected on 13 September 1999 at Camp Bonifas were positive for P. vivax 247. PMID- 11931267 TI - Isolation of west nile and sindbis viruses from mosquitoes collected in the Nile Valley of Egypt during an outbreak of Rift Valley fever. AB - As part of an evaluation of potential vectors of arboviruses during a Rift Valley fever (RVF) outbreak in the Nile Valley of Egypt in August 1993, we collected mosquitoes in villages with known RVF viral activity. Mosquitoes were sorted to species, pooled, and processed for virus isolation both by intracerebral inoculation into suckling mice and by inoculation into cell culture. A total of 33 virus isolates was made from 36,024 mosquitoes. Viruses were initially identified by indirect fluorescent antibody testing and consisted of 30 flaviviruses (all members of the Japanese encephalitis complex, most probably West Nile [WN] virus) and three alphaviruses (all members of western equine encephalitis complex, most probably Sindbis). The identity of selected viruses was confirmed by reverse transcriptase-polymerase chain reaction and sequencing. Culex antennatus (Becker) and Culex perexiguus Theobald accounted for five (17%) and 23 (77%) of the WN virus isolations, respectively. Despite isolation of viruses from 32 pools of mosquitoes (both WN and Sindbis viruses were isolated from a single pool), RVF virus was not isolated from these mosquitoes, even though most of them are known competent vectors collected during an ongoing RVF outbreak. Thus, it should be remembered, that even during a known arbovirus outbreak, other arboviruses may still be circulating and causing disease. PMID- 11931268 TI - Keys to the Anopheline mosquitoes (Diptera: Culicidae) of Pakistan. AB - This article provides taxonomic keys for the identification of the fourth-instar larvae and females of 24 species of anopheline mosquitoes (seven species in subgenus Anopheles and 17 species in subgenus Cellia) recorded from Pakistan. The keys are based on literature sources as well as the examination of field and museum collections. PMID- 11931270 TI - Analysis of northern distribution of Aedes albopictus (Diptera: Culicidae) in Japan by geographical information system. AB - Aedes albopictus (Skuse), a mosquito vector of the dengue fever virus, is prevalent in Japan, distributed widely in Honshu Island with its northern limits between latitude 38 degrees to 40 degrees north. The factors affecting distribution of the species in the northern part of Japan were studied using the geographical information system (GIS). During 1998-2000, larval surveillance was carried out in 26 urban and rural areas in the Tohoku district, in the northern part of Honshu Island, by collecting larvae from artificial and natural habitats. Climatological analysis, using the GIS, showed that the following conditions accounted for the current distribution of Ae. albopictus: an annual mean temperature higher than 11 degrees C and a mean temperature of the coldest month, January, higher than -2 degrees C. A period with temperature above 11 degrees C in the confirmed area of the mosquito successively continues for more than 186 d per year. The accumulated temperature calculated from a temperature of 11 degrees C, which may be close to the developmental zero of Ae. albopictus, was over 1,350 degree-days. The relationship between the beginning of short-daylength, inducing egg diapause, and the monthly mean temperature during September and October necessary for successful larval development in the Tohoku district is also discussed. We also show the relationship between the current distribution of Ae. albopictus and the annual mean temperature in the United States. From these results it is predicted that Ae. albpictus will be established in some cities in northeast United States. PMID- 11931269 TI - Morphology of the antenna of Dermatobia hominis (Diptera: Cuterebridae) based on scanning electron microscopy. AB - Sensilla of the antennae of male and female Dermatobia hominis were studied by scanning electron microscopy. The images obtained were interpreted according to the scientific literature referring to the sensory structures of insects. Sensilla of the "long bristle" type and smaller spines of the "microtrichia" type were found in different numbers and patterns of distribution on the scape and pedicel. Coeloconic sensilla with longitudinal cuticular furrows were observed on the female flagellum, as well as two varieties of basiconic sensilla: a large one surrounded by pointed foliaceous structures and a smaller form implanted on a raised cuticular process. The larger type of basiconic sensilla was also observed on the flagellum of the male antenna, as well as a variety of coeloconic sensilla with apical dilatations. Trichoid and chaetic sensilla were encountered in greater numbers on the arista of females, with the former type predominating. Coeloconic sensilla were observed on the arista of both sexes, as well as sensilla of the "long bristle" and styloconic types exclusively in males. Adult D. hominis were observed to possess sensory structures with chemoreceptory, thermo-hygroreceptory and mechanoreceptory functions on their antennae. These results could facilitate the identification of the chemoreceptors by electrophysiological techniques. The sexual dimorphism noted for the antennae constitutes a new criterion for distinguishing between the sexes of D. hominis PMID- 11931271 TI - Lack of manipulation of Rhodnius prolixus (Hemiptera: Reduviidae) vector competence by Trypanosoma cruzi. AB - Behavioral implications of Trypanosoma cruzi Chagas infection in Rhodnius prolixus Stal were observed. Feeding and defecation behaviors of infected versus uninfected insects were assessed on an artificial membrane-feeding system and on live guinea pigs. Based on a defecation index, fifth instars were the most efficient vectors, followed by adult females, fourth instars, and adult males. Bugs fasted for longer periods (5-6 mo) took smaller blood meals but defecated significantly earlier than bugs fasted for shorter periods (2-3 mo). Multiple blood feeding, degree of fasting, life stage, T cruzi infection, and gender affected the vector potential of R. prolixus. Our data indicate that T. cruzi and R. prolixus have not coevolved to facilitate the transmission of T. cruzi, which suggests that this parasite-host relationship may be relatively recent. PMID- 11931272 TI - Assessment of parasitism of house fly and stable fly (Diptera: Muscidae) pupae by pteromalid (Hymenoptera: Pteromalidae) parasitoids using a polymerase chain reaction assay. AB - The internal transcribed spacer (ITS) regions of the ribosomal DNA of house flies, Musca domestica L., the stable flies, Stomoxys calcitrans (L.), and four parasitoid species in the genus Muscidifurax (Hymenoptera: Pteromalidae) were characterized to develop a method based on the polymerase chain reaction (PCR) to better define the role of pteromalid parasitism of pupae of the house fly and stable fly. Two parasitoid-specific primers were designed to anneal to the 5' end of the 5.8S rRNA gene in the parasitoid species. When paired with a universal primer at the 3' end of the 18S rRNA, the primers amplified the target ITS1 region in 10 pteromalid species. PCR allowed detection of parasitoid DNA within 24 h after females of Spalangia endius Walker oviposited into house fly puparia. PCR failed to amplify parasitoid DNA or detect parasitism in puparia that were exposed to parasitoid oviposition, allowed to develop 7 d, then killed by freezing and held at 20-24 degrees C for 4 d to allow DNA degradation. Digestion of the PCR products with restriction enzymes produced restriction fragment length polymorphisms that allowed identification of individual parasitoid species. Significantly greater levels of parasitism (P < 0.05) were detected by PCR for two of the five field collection dates in 1997. On the dates when PCR detected higher levels of parasitism than estimates provided by emergence of adult insects from samples taken at Feedlot M in 1997, more than 65% of all puparia in the emergence samples failed to produce an adult insect. Three puparia collected in 1997 produced double PCR bands that corresponded to PCR band sizes of Muscidifurax spp. and Spalangia sp., possibly indicating multiple parasitism or hyperparasitism. PMID- 11931273 TI - Cellular-mediated reactions to foreign organisms inoculated into the hemocoel of Anopheles albimanus (Diptera: Culicidae). AB - The immune response against different organisms and particles inoculated in the hemocoel of female Anopheles albimanus Wiedemann was investigated. Histological and ultrastructural observations indicated that melanization and hemocyte type participation varied according to the particles inoculated. The initial responses against heat-killed Microccocus lysodeikticus and Escherichia coli included hemocyte lysis and melanization whereas the response to heat-killed Saccharomyces cerevisiae was only cellular, and an initial melanization of Sephadex G-25 (neutral charged) beads was followed by the formation of cellular aggregates. After 24 h, hemocytes were involved in all terminal encapsulation events. Plasmodium vivax Grassi and Feletti formalin-fixed sporozoites induced a weak response. Cellular aggregates were observed 1 h postinoculation, but participating hemocytes could not be identified because of the extensive cellular damage and lysis. Sporozoites were also observed in the core of these aggregates, mixed with cell debris and free in the hemolymph. The effect on the inoculated particles was also different-S. cerevisiae was encapsulated only by hemocytes, whereas M. lysodeikticus was lysed and E. coli was phagocytosed by plasmatocytes. These results indicate that hemocytes are important components in the immune response in An. albimanus. PMID- 11931274 TI - Spatial and habitat distribution of Anopheles gambiae and Anopheles arabiensis (Diptera: Culicidae) in Banambani village, Mali. AB - We studied the larval distribution and composition of Anopheles arabiensis Patton, An. gambiae s.s. Giles, and its forms, among local habitats; and their association with the adults between these habitats in Banambani village, Mali during the mid-rainy seasons of 1997-1999. For species and form identification we used polymerase chain reaction (PCR) and PCR-restriction fragment-length polymorphism (RFLP). Differences among species in the distribution of larvae were observed in 1998, but not in 1997 or 1999, although they were on the borderline of statistical significance. Differences among the M and S molecular forms were statistically significant in 1999 when rainfall was high, but not in the two prior, drier sampling periods. Combining all information into the Fisher multiple comparisons test, there were statistically significant differences between species and molecular forms during the 3-yr study period. Hybrid larvae between the M and S forms were observed (0.57%), the first such observation to our knowledge. In spite of differences among larval distribution, no differences of adult species composition were observed among habitats. Factors that influence the distributions of An. gambiae larval populations are discussed. PMID- 11931275 TI - Analysis of the population structure of Anopheles funestus (Diptera: Culicidae) from western and coastal Kenya using paracentric chromosomal inversion frequencies. AB - In total, 324 Anopheles funestus Giles specimens collected from seven houses in western Kenya and seven in coastal Kenya were scored for their paracentric chromosomal inversions with the aim of determining the level of genetic differentiation based on these inversions. Houses in each area were within a 2-km radius. The two areas are approximately 700 km apart. Only inversions 2a, 3a, 3b, and 5a were found to be polymorphic. Levels of polymorphism varied greatly between inversions. There were no significant deviations from Hardy-Weinberg expectations for samples from individual houses at one site or when data for houses in each area were pooled. Overall, the level of differentiation between western and coastal Kenya was significant, suggesting that the two populations are genetically isolated. Results based on inversion 2a alone were, however, not consistent with this conclusion. Founder effects and selection against the 2a inversion are discussed as possible explanations for this discrepancy. PMID- 11931276 TI - Melanization of plasmodium falciparum and C-25 sephadex beads by field-caught Anopheles gambiae (Diptera: Culicidae) from southern Tanzania. AB - The melanization responses of field-captured Anopheles gambiae s.l. toward oocysts of the malaria parasite Plasmodium falciparum or negatively charged (C 25) Sephadex beads were determined. Only two of 431 infected mosquitoes harboured melanized oocysts. However, 90% of field-captured mosquitoes melanized C-25 Sephadex beads. The effects of age, glucose concentration and blood meal on the melanization response of an An gambiae s.s. laboratory colony toward C-25 beads were also assayed. All newly emerged females (which did not blood-feed) melanized the beads. By 4 d postemergence, there was a marked reduction in melanization response, particularly among those mosquitoes that had not blood fed. A blood meal, however, taken by 4-d-old mosquitoes increased their immune response as did high glucose concentrations in the nonblood-fed group. These data indicate that C 25 Sephadex beads can estimate the general strength of An. gambiae's immune response. However, C-25 beads do not accurately model An. gambiae's susceptibility to P falciparum oocysts in natural populations. To the best of our knowledge, this is the first report of field refractoriness in An. gambiae s.l. PMID- 11931277 TI - Comparsion of selected growth media for culturing Serratia marcescens, Aeromonas sp., and Pseudomonas aeruginosa as pathogens of adult Stomoxys calcitrans (Diptera: Muscidae). AB - Stable flies, Stomoxys calcitrans (L.), were orally infected with Aeromonas sp., Pseudomonas aeruginosa (Schroeter), and Serratia marcescens Bizio that were cultured on egg-yolk media, nutrient broth, and fly egg media. Aeromonas and Serratia caused mortality when the bacteria were originally grown on egg-yolk medium. Pseudomonas was equally lethal regardless of the media on which it was cultured. A wild isolate of Aeromonas caused greater death than an isolate that had been passed through host flies and had been reisolated from killed flies. Mortality increased with bacterial dose for all species. Aeromonas and Serratia caused mortality within several days after ingestion, whereas Pseudomonas caused a gradual increase in mortality 3-7 d after ingestion. The pathologic activity of Aeromonas and Serratia required extracellular products produced when cells were grown in egg yolk medium. Aeromonas required both supernatant and cells from egg yolk medium, wereas Serratia required supernatant from egg yolk medium and cells from either nutrient broth or egg yolk medium. Mortality due to ingestion of Aeromonas was correlated with the presence of enzymes that cause alpha- and beta hemolysis, while mortality following ingestion of Serratia was associated with alpha-hemolysins, elastases, and chitinases. PMID- 11931278 TI - Enhancement or modulation of the vector competence of Ochlerotatus vigilax (Diptera: Culicidae) for ross river virus by temperature. AB - Two different doses of Ross River virus (RR) were fed to Ochlerotatus vigilax (Skuse), the primary coastal vector in Australia; and blood engorged females were held at different temperatures up to 35 d. After ingesting 10(4.3) CCID50/mosquito, mosquitoes reared at 18 and 25 degrees C (and held at the same temperature) had higher body remnant and head and salivary gland titers than those held at 32 degrees C. although infection rates were comparable. At 18, 25, and 32 degrees C, respectively, virus was first detected in the salivary glands on days 3, 2, and 3. Based on a previously demonstrated 98.7% concordance between salivary gland infection and transmission, the extrinsic incubation periods were estimated as 5, 4, and 3 d, respectively, for these three temperatures. When Oc. vigilax reared at 18, 25, or 32 degrees C were fed a lower dosage of 10(3.3) CCID50 RR/mosquito, and assayed after 7 d extrinsic incubation at these (or combinations of these) temperatures, infection rates and titers were similar. However, by 14 d, infection rates and titers of those reared and held at 18 and 32 degrees C were significantly higher and lower, respectively. However, this process was reversible when the moderate 25 degrees C was involved, and intermediate infection rates and titers resulted. These data indicate that for the strains of RR and Oc. vigilax used, rearing temperature is unimportant to vector competence in the field, and that ambient temperature variations will modulate or enhance detectable infection rates only after 7 d extrinsic incubation. Because of the short duration of extrinsic incubation, however, this will do little to influence RR epidemiology, because by this time some Oc. vigilax could be seeking their third blood meal, the latter two being infectious. PMID- 11931279 TI - Well- and ill-defined measures of everyday cognition: relationship to older adults' intellectual ability and functional status. AB - The present study examined 2 approaches to the measurement of everyday cognition in older adults. Measures differing in the degree of structure offered for solving problems in the domains of medication use, financial management, and food preparation and nutrition were administered to a sample of 130 community-dwelling older adults ranging in age from 60 to 90 (M = 73 years, SD = 7.02 years). Well defined and ill-defined everyday problem-solving measures, which varied in the amount of means-end-related information provided to participants, were used. The study found that (a) well- and ill-defined measures were moderately interrelated, (b) the 2 approaches were differentially related to basic cognitive abilities, and (c) together the 2 approaches explained over half of the variance in older adults' everyday instrumental functioning and were in fact better predictors of everyday functioning than traditional psychometric cognitive measures. Discussion focuses on the differential importance of both methods for assessing older adults' everyday cognitive functioning. PMID- 11931281 TI - Time counts: future time perspective, goals, and social relationships. AB - On the basis of postulates derived from socioemotional selectivity theory, the authors explored the extent to which future time perspective (FTP) is related to social motivation, and to the composition and perceived quality of personal networks. Four hundred eighty German participants with ages ranging from 20 to 90 years took part in the study. In 2 card-sort tasks, participants indicated their partner preference and goal priority. Participants also completed questionnaires on personal networks and social satisfaction. Older people, as a group, perceived their future time as more limited than younger people. Individuals who perceived future time as being limited prioritized emotionally meaningful goals (e.g., generativity, emotion regulation), whereas individuals who perceived their futures as open-ended prioritized instrumental or knowledge-related goals. Priority of goal domains was found to be differently associated with the size, composition, and perceived quality of personal networks depending on FTP. Prioritizing emotion-regulatory goals was associated with greater social satisfaction and less perceived strain with others when participants perceived their future as limited. Findings underscore the importance of FTP in the self regulation of social relationships and the subjective experience associated with them. PMID- 11931280 TI - Age differences and similarities in the correlates of depressive symptoms. AB - The authors investigated whether several life events or concerns were differentially related to depressive symptoms across 3 adult age groups (young adults, middle-aged adults, and older adults). They examined the relationships of 2 measures of depressive symptoms to work status and satisfaction, relationship status and satisfaction, loneliness, recent losses, parenting strain, and caregiving. Some differences between age groups in these relationships were found. Yet, most results suggested that, although the frequency with which people experience specific life events or concerns varies across the adult life span, the relationships between these events or concerns and depressive symptoms are similar across age groups. PMID- 11931282 TI - Age-related slowing in face and name recognition: evidence from event-related brain potentials. AB - Age-related slowing in recognizing famous names and faces was investigated with event-related brain potentials (ERPs). In a group of young adults, item repetition induced early (220-340 ms) and late (400-700 ms) ERP modulations, apparently signaling the access to, respectively, domain-specific representations of faces and names and domain-general semantic knowledge about the persons. These repetition effects and other ERP components were then used as process-specific time markers in middle-aged and elderly participants. For both faces and names, the elderly participants' responses were slowed, but repetition priming in reaction times was not. The ERP latencies suggested that most of the age-related slowing occurred in the access to domain-specific representations and during response decision, whereas sensory and perceptual processing was largely spared. PMID- 11931283 TI - Longitudinal analysis of midlife generativity, intergenerational roles, and caregiving. AB - Items from the California Adult Q-Sort (CAQ) were used to assess psychosocial generativity (E. H. Erikson, 1950) in a sample of educated women at midlife. CAQ scores measured at age 43 demonstrated convergent validity with an inventory measure of generativity assessed at age 53. According to other longitudinal analyses, women who attained a generative stance at age 43 reported greater investment 10 years later in intergenerational roles (e.g., daughter, mother) but not nonintergenerational ones (e.g., friend, sister). Generative women also reported less subjective burden in caring for aging parents and more knowledge about community elder care programs. Further evidence suggests that participants felt embedded in a reciprocal caregiving network in which they themselves were the recipients of care. PMID- 11931284 TI - A medication screening tool for cognitive aging researchers. AB - A Web-based medication screening tool that provides researchers with information about side effects associated with medications commonly used by older participants is described. This tool can be used for research purposes to better separate the effects of normal, healthy aging processes from the deleterious effects often associated with medication usage. Researchers can use this tool, called SMART (Screening Medications: Aging Research Taxonomy), to (a) obtain information pertaining to the cognitive, sensory, and motor side effects associated with specific medications and (b) screen medications in their research for side effect severity. The Web address for the tool is www.psychology.gatech.edu/SMART PMID- 11931286 TI - Introduction to the special section on aging, cognition, and neuroimaging. AB - In response to the recent increase in the number of laboratories engaged in the use of functional and structural neuroimaging to study cognitive aging, this special section has been compiled to serve as an entry into this area for the readers of Psychology and Aging. These articles are representative of the field and cover many of the issues faced by researchers in this area. This introduction presents some background into the techniques that are used and provides an overview of the articles. PMID- 11931285 TI - Aging and attentional guidance during visual search: functional neuroanatomy by positron emission tomography. AB - Positron emission tomography (PET) was used to examine adult age differences in neural activation during visual search. Target detection was less accurate for older adults than for younger adults, but both age groups were successful in using color to guide attention to a subset of display items. Increasing perceptual difficulty led to greater activation of occipitotemporal cortex for younger adults than for older adults, apparently as the result of older adults maintaining higher levels of activation within the easier task conditions. The results suggest that compensation for age-related decline in the efficiency of occipitotemporal cortical functioning was implemented by changes in the relative level of activation within this visual processing pathway, rather than by the recruitment of other cortical regions. PMID- 11931287 TI - Differences in the functional neuroanatomy of inhibitory control across the adult life span. AB - Inhibitory control, the ability to suppress irrelevant or interfering stimuli, is a fundamental cognitive function that deteriorates during aging, but little is understood about the bases of decline. Thus, we used event-related functional magnetic resonance imaging (fMRI) to study inhibitory control in healthy adults aged 18 to 78. Activation during "successful inhibition" occurred predominantly in right prefrontal and parietal regions and was more extensive, bilaterally and prefrontally, in the older groups. Presupplementary motor area was also more active in poorer inhibitory performers. Therefore, older adults activate areas that are comparable to those activated by young adults during inhibition, as well as additional regions. The results are consistent with a compensatory interpretation and extend the aging neuroimaging literature into the cognitive domain of inhibition. PMID- 11931288 TI - The effects of encoding task on age-related differences in the functional neuroanatomy of face memory. AB - Age-related differences in brain activity mediating face recognition were examined using positron emission tomography. Participants encoded faces using a pleasant-unpleasant judgment, a right-left orientation task, and intentional learning. Scans also were obtained during recognition. Both young and old groups showed signficant effects of encoding task on recognition accuracy, but older adults showed reduced accuracy overall. Increased brain activity in older adults was similar to that seen in young adults during conditions associated with deeper processing, but was reduced during the shallow encoding and recognition conditions. Left prefrontal activity was less in older adults during encoding, but greater during recognition. Differential correlations of brain activity and behavior were found that suggest older adults use unique neural systems to facilitate face memory. PMID- 11931289 TI - Age-related differences in the course of cognitive skill acquisition: the role of regional cortical shrinkage and cognitive resources. AB - This study examined the impact of age-related differences in regional cerebral volumes and cognitive resources on acquisition of a cognitive skill. Volumes of brain regions were measured on magnetic resonance images of healthy adults (aged 22-80). At the early stage of learning to solve the Tower of Hanoi puzzle, speed and efficiency were associated with age, prefrontal cortex volume, and working memory. A similar pattern of brain-behavior associations was observed with perseveration measured on the Wisconsin Card Sorting Test. None of the examined structural brain variables were important at the later stages of skill acquisition. When hypertensive participants were excluded, the effect of prefrontal shrinkage on executive aspects of performance was no longer significant, but the effect of working memory remained. PMID- 11931290 TI - Hemispheric asymmetry reduction in older adults: the HAROLD model. AB - A model of the effects of aging on brain activity during cognitive performance is introduced. The model is called HAROLD (hemispheric asymmetry reduction in older adults), and it states that, under similar circumstances, prefrontal activity during cognitive performances tends to be less lateralized in older adults than in younger adults. The model is supported by functional neuroimaging and other evidence in the domains of episodic memory, semantic memory, working memory, perception, and inhibitory control. Age-related hemispheric asymmetry reductions may have a compensatory function or they may reflect a dedifferentiation process. They may have a cognitive or neural origin, and they may reflect regional or network mechanisms. The HAROLD model is a cognitive neuroscience model that integrates ideas and findings from psychology and neuroscience of aging. PMID- 11931291 TI - Individual acoustic variation in Belding's ground squirrel alarm chirps in the High Sierra Nevada. PMID- 11931292 TI - Optical and atomic force microscopic studies on sonoporation. PMID- 11931293 TI - Measurement of the scattering of a Lamb wave by a through hole in a plate. AB - Flexural waves propagating in an aluminum plate containing a circular hole are studied. In the experiments the first antisymmetric Lamb wave mode A0 is excited selectively by a piezoelectric transducer. The scattered field around a circular cavity is measured pointwise using a heterodyne laser interferometer. The measurements are compared with theoretical calculations. Different approximate analytical approaches, employing Kirchhoff and Mindlin types of plate theories to describe the scattered field, are used. Good agreement between the experimental data and the analytical solutions is found within the ranges of validity of the different models. Introduction of a small imperfection, like a notch, at the boundary of the cavity changes the measured scattered field PMID- 11931294 TI - On the selection of loads in the multiload method for measuring the acoustic source parameters of duct systems. AB - The in-duct source can be characterized by two acoustical parameters such as the source strength and the source impedance, which permit the prediction of radiated sound pressure or insertion loss of the whole duct system. One-port acoustic characteristics of an in-duct source can be measured by the multiload method using an overdetermined set of open pipes or side-branch pipes with different lengths as applied loads. The input data, viz. load pressure and load impedance, are usually contaminated by measurement error in the actual measurements, which result in errors in the calculated source parameters. In this paper, the effects of the errors in the input data on the results have been studied numerically, varying the number of loads and their impedances in order to determine what combination of the loads will yield the best result. It is noted that, frequently, only a set of open pipes is used when applying the multiload method to the internal combustion engine sources. A set of pipe lengths, which cause the calculated results to be least sensitive to the input data error, can be found when using open pipe loads. The present work is intended to produce guidelines for preparing an appropriate load set in order to obtain accurate source properties of fluid machines. PMID- 11931295 TI - An approximation to the far field and directivity of elastic wave transducers. AB - The far field of surface mounted elastic wave transducers, as used in Non Destructive Testing (NDT), is analyzed in order to understand the mechanisms that determine the directivity of these devices. To model the physical configuration, a moving distribution of normal traction, acting at the free surface of an elastic half-space, is used. The exact field expression consists of a spatial superposition of the elementary waves emitted by the hypothetical point sources that make up the finite source. The current aim is to derive approximate far field expressions that are physically instructive and numerically fast. This is achieved through a paraxial approximation of the elementary waves and a Taylor approximation of their arrival times. The approximate expressions thus obtained, consist of a time convolution of a paraxial wave and a transfer function. The latter represents the influence of the source geometry. Besides elucidating the mechanisms of directivity, the analysis renders a rule of thumb for synthesizing a source with a desired directivity pattern. Numerical experiments show that the approximations reduce the computational effort by a factor of 1000, and that the accuracy of the approximate far field and directivity proves excellent at distances beyond a few times the characteristic source size. PMID- 11931296 TI - Rotational aerophones. AB - Free rotational aerophones such as the bullroarer, which consists of a wooden slat whirled around on the end of a string, and which emits a loud pulsating roar, have been used in many ancient and traditional societies for ceremonial purposes. This article presents an experimental and theoretical investigation of this instrument. The aerodynamics of rotational behavior is elucidated, and relates slat rotation frequency to slat width and velocity through the air. Analysis shows that sound production is due to generation of an oscillating rotating dipole across the slat, the role of the vortices shed by the slat being relatively minor. Apparent discrepancies between the behavior of a bullroarer slat and a slat mounted on an axle in a wind tunnel are shown to be due to viscous friction in the bearings of the wind-tunnel experiment. PMID- 11931297 TI - Three-dimensional modeling of acoustic backscattering from fluid-like zooplankton. AB - Scattering models that correctly incorporate organism size and shape are a critical component for the remote detection and classification of many marine organisms. In this work, an acoustic scattering model has been developed for fluid-like zooplankton that is based on the distorted wave Born approximation (DWBA) and that makes use of high-resolution three-dimensional measurements of the animal's outer boundary shape. High-resolution computerized tomography (CT) was used to determine the three-dimensional digitizations of animal shape. This study focuses on developing the methodology for incorporating high-resolution CT scans into a scattering model that is generally valid for any body with fluid like material properties. The model predictions are compared to controlled laboratory measurements of the acoustic backscattering from live individual decapod shrimp. The frequency range used was 50 kHz to 1 MHz and the angular characteristics of the backscattering were investigated with up to a 1 degree angular resolution. The practical conditions under which it is necessary to make use of high-resolution digitizations of shape are assessed. PMID- 11931298 TI - An ensemble source spectra model for merchant ship-radiated noise. AB - This paper presents an evaluation of the classical model for determining an ensemble of the broadband source spectra of the sound generated by individual ships and proposes an alternate model to overcome the deficiencies in the classical model. The classical model, proposed by Ross [Mechanics of Underwater Noise (Pergamon, New York, 1976)] postulates that the source spectrum for an individual ship is proportional to a baseline spectrum with the constant of proportionality determined by a power-law relationship on the ship speed and length. The model evaluation, conducted on an ensemble of 54 source spectra over a 30-1200-Hz to 1200-Hz frequency band, shows that this assumption yields large rms errors in the broadband source level for the individual ships and significantly overestimates the variability in the source level across the ensemble of source spectra. These deficiencies are a consequence of the negligible correlation between the source level and the ship speed and the source level and the ship length. The alternate model proposed here represents the individual ship spectra by a modified rational spectrum where the poles and zeros are restricted to the real axis and the exponents of the terms are not restricted to integer values. An evaluation of this model on the source spectra ensemble indicates that the rms errors are significantly less than those obtained with any model where the frequency dependence is represented by a single baseline spectrum. Furthermore, at high frequencies (400 to 1200 Hz), a single-term rational spectrum model is sufficient to describe the frequency dependence and, at the low frequencies (30 to 400 Hz), there is only a modest reduction in the rms error for a higher order model. Finally, a joint probability density on the two parameters of the single term model based on the measured histograms of these parameters is proposed. This probability density provides a mechanism for generating an ensemble of ship spectra. PMID- 11931299 TI - Identification of complex stiffness tensor from waveform reconstruction. AB - An inverse method is proposed in order to determine the viscoelastic properties of composite-material plates from the plane-wave transmitted acoustic field. Analytical formulations of both the plate transmission coefficient and its first and second derivatives are established, and included in a two-step inversion scheme. Two objective functions to be minimized are then designed by considering the well-known maximum-likelihood principle and by using an analytic signal formulation. Through these innovative objective functions, the robustness of the inversion process against high level of noise in waveforms is improved and the method can be applied to a very thin specimen. The suitability of the inversion process for viscoelastic property identification is demonstrated using simulated data for composite materials with different anisotropy and damping degrees. A study of the effect of the rheologic model choice on the elastic property identification emphasizes the relevance of using a phenomenological description considering viscosity. Experimental characterizations show then the good reliability of the proposed approach. Difficulties arise experimentally for particular anisotropic media. PMID- 11931300 TI - Determination of the elastic constants of a composite plate using wavelet transforms and neural networks. AB - An inverse method based on a combination of the wavelet transform and artificial neural networks is presented. The method is used to recover the elastic constants of a fiber-reinforced composite plate from experimental measurements of ultrasonic Lamb waves generated and detected with lasers. In this method, the elastic constants are not recovered from the dispersion curves but rather directly from the measured waveforms. Transient waveforms obtained by numerical simulations for different elastic constants are used as input to train the neural network. The wavelet transform is used to extract the eigenvectors from the Lamb wave signals to simplify the structure of the neutral network. The eigenvectors are then introduced into a multilayer internally recurrent neural network with a back-propagation algorithm. Finally, experimental waveforms recoded on a titanium graphite composite plate are used as input to recover the elastic constants of the material. PMID- 11931301 TI - Adaptive tuning of an electrodynamically driven thermoacoustic cooler. AB - The commercial development of thermoacoustic coolers has been hampered in part by their low efficiencies compared to vapor compression systems. A key component of electrodynamically driven coolers is the electromechanical transducer, or driver. The driver's electroacoustic transduction efficiency, defined as the ratio of the acoustic power delivered to the working gas by the moving piston and the electrical power supplied, must be maintained near its maximum value if a high overall system efficiency is to be achieved. Modeling and experiments have shown that the electroacoustic efficiency peaks sharply near the resonance frequency of the electro-mechano-acoustic system. The optimal operating frequency changes as the loading condition changes, and as the properties of the working gas vary. The driver efficiency may thus drop significantly during continuous operation at a fixed frequency. In this study, an on-line driver efficiency measurement scheme was implemented. It was found that the frequency for maximum electroacoustic efficiency does not precisely match any particular resonance frequency, and that the efficiency at resonance can be significantly lower than the highest achievable efficiency. Therefore, a direct efficiency measurement scheme was implemented and validated using a functional thermoacoustic cooler. An adaptive frequency-tuning scheme was then implemented. Experiments were performed to investigate the effectiveness of the control scheme to maintain the maximum achievable driver efficiency for varying operating conditions. PMID- 11931302 TI - Single half-wavelength ultrasonic particle filter: predictions of the transfer matrix multilayer resonator model and experimental filtration results. AB - The quantitative performance of a "single half-wavelength" acoustic resonator operated at frequencies around 3 MHz as a continuous flow microparticle filter has been investigated. Standing wave acoustic radiation pressure on suspended particles (5-microm latex) drives them towards the center of the half-wavelength separation channel. Clarified suspending phase from the region closest to the filter wall is drawn away through a downstream outlet. The filtration efficiency of the device was established from continuous turbidity measurements at the filter outlet. The frequency dependence of the acoustic energy density in the aqueous particle suspension layer of the filter system was obtained by application of the transfer matrix model [H. Nowotny and E. Benes, J. Acoust. Soc. Am. 82, 513-521 (1987)]. Both the measured clearances and the calculated energy density distributions showed a maximum at the fundamental of the piezoceramic transducer and a second, significantly larger, maximum at another system's resonance not coinciding with any of the transducer or empty chamber resonances. The calculated frequency of this principal energy density maximum was in excellent agreement with the optimal clearance frequency for the four tested channel widths. The high-resolution measurements of filter performance provide, for the first time, direct verification of the matrix model predictions of the frequency dependence of acoustic energy density in the water layer. PMID- 11931303 TI - Measurement of an aeroacoustic dipole using a linear microphone array. AB - It is shown that the standard beamformer technique is inadequate for both the source location and the measurement of a simple dipole and that this is due to the assumption of monopole propagation in the calculation of the phase weights used to steer the focus of the array. A numerical simulation is used to illustrate the problem and to develop a correction to the signal processing algorithm to account for the dipole propagation characteristic. This is then applied to array measurements for an aeroacoustic dipole produced by a cylinder in a cross flow. The resulting source map and the beamformed spectrum are shown to give a true representation of the source energy and frequency content. A secondary effect of this correction is that the array becomes insensitive to other source types so that in addition to acting as a spatial filter, the array can perform as a source filter. This work also demonstrates how an array measurement can be misinterpreted if applied without consideration of the source mechanism. PMID- 11931304 TI - Estimation of broadband acoustic power radiated from a turbulent boundary layer driven reinforced finite plate section due to rib and boundary forces. AB - Previous papers considered an infinite fluid-loaded plate with parallel line attachments, driven by a wave-number-white pressure excitation invariant in the direction of an attachment, and established the conditions and procedure for estimating the broadband radiated power by assuming the ribs to radiate independently. This paper applies those results to a finite rectangular ribbed plate, and extends the methodology to include the contribution of the plate's boundary support forces to the radiation and the consideration of excitation that varies in the direction parallel to the ribs. The approach is relevant to problems of sound radiation by underwater stiffened steel plates driven by turbulent boundary layer (TBL) pressures, and is also applicable to stiffened circular cylindrical shells when the response is dominated by bending. Comparisons of sample calculations with results of rigorous models validate the approximation. PMID- 11931306 TI - Backscattering of transients by tilted truncated cylindrical shells: time frequency identification of ray contributions from measurements. AB - Impulse backscattering measurements by a thick-walled finite cylindrical shell are examined in the time-frequency domain to identify and characterize individual ray contributions from generalized Lamb waves excited on the shell. Previous experiments and analysis in the frequency-aspect angle domain [S. F. Morse et al., J. Acoust. Soc. Am. 103, 785-794 (1998)] indicate that large backscattering enhancements occur in the midfrequency region for the shell tilted at large angles. Presently this experimental data is examined in the time-frequency domain for selected angles of incidence. Individual ray contributions are evident and their evolution over aspect angle is discussed. The most prominent contribution is due to the meridional ray of the a0 leaky Lamb wave. This feature distinctly highlights the truncation of the shell and is found over a range of aspect angles spanning 200 degrees for the frequencies examined. Also observed are periodic features corresponding to end-reflected helical waves of the a0-. These scattering features are significantly different from those reported for thin walled finite cylinders at low frequencies. The present results may be useful for target identification and localization and as a comparison tool for high frequency computational scattering models. PMID- 11931305 TI - Effect of phase on discomfort caused by vertical whole-body vibration and shock- experimental investigation. AB - An experimental study has investigated the effect of "phase" on the subjective responses of human subjects exposed to vertical whole-body vibration and shock. The stimuli were formed from two frequency components: 3 and 9 Hz for continuous vibrations and 3 and 12 Hz for shocks. The two frequency components, each having 1.0 ms(-2) peak acceleration, were combined to form various waveforms. The effects of the vibration magnitude on the discomfort caused by the input stimuli were also investigated with both the continuous vibrations and the shocks. Various objective measurements of acceleration and force at the seat surface, the effects of different frequency weightings and second and fourth power evaluations were compared with judgments of the discomfort of the stimuli. It was found that a 6% to 12% increase in magnitude produced a statistically significant increase in discomfort with both the continuous vibrations and the shocks. Judgments of discomfort caused by changes in vibration magnitude were highly correlated with all of the objective measurements used in the study. The effects on discomfort of the phase between components in the continuous vibrations were not statistically significant, as predicted using evaluation methods with a power of 2. However, small changes in discomfort were correlated with the vibration dose value (VDV) of the Wb frequency-weighted acceleration. The effect of phase between frequency components within the shocks was statistically significant, although no objective measurement method used in the study was correlated with the subjective judgments. PMID- 11931307 TI - Broadband radiation modes: estimation and active control. AB - In this paper we give a formulation of the most efficiently radiating vibration patterns of a vibrating body, the radiation modes, in the time domain. The radiation modes can be used to arrive at efficient weighting schemes for an array of sensors in order to reduce the controller dimensionality. Because these particular radiation modes are optimum in a broadband sense, they are termed broadband radiation modes. Methods are given to obtain these modes from measured data. The broadband radiation modes are used for the design of an actuator array in a feedback control system to reduce the sound power radiated from a plate. Three methods for the design of the actuator are compared, taking into account the reduction of radiated sound power in the controlled frequency range, but also the possible increase of radiated sound power in the uncontrolled frequency range. PMID- 11931308 TI - Modeling of the human middle ear using the finite-element method. AB - In this study, a three-dimensional finite-element model (FEM) of the human middle ear was established, including features of the middle ear which were not considered in the previous model, i.e., the ligaments, tendons, I-S joint, loading of the cochlea, external auditory meatus (EAM), middle-ear cavities, etc. The unknown mechanical properties of these parts and the boundary conditions were determined so that the impedance obtained from the FEM analysis resembled the measurement values. The validity of this model was confirmed by comparing the motion of the tympanic membrane and ossicles obtained by this model with the measurement data, and the effects of the newly considered features on the numerically obtained results were examined. By taking the ligaments and tendons into account and assuming that the cochlea acts as a damper, with this model it was possible to realistically reproduce complex ossicular chain movement. It was found that the middle-ear cavities did not affect the vibration mode of the tympanic membrane. Although the EAM enhanced the sound pressure applied to the tympanic membrane compared with that at the entrance of the EAM, the pressure distribution on the surface of the tympanic membrane was not affected by the EAM. PMID- 11931309 TI - Frequency specificity of chirp-evoked auditory brainstem responses. AB - This study examines the usefulness of the upward chirp stimulus developed by Dau et al. [J. Acoust. Soc. Am. 107, 1530-1540 (2000)] for retrieving frequency specific information. The chirp was designed to produce simultaneous displacement maxima along the cochlear partition by compensating for frequency-dependent traveling-time differences. In the first experiment, auditory brainstem responses (ABR) elicited by the click and the broadband chirp were obtained in the presence of high-pass masking noise, with cutoff frequencies of 0.5, 1, 2, 4, and 8 kHz. Results revealed a larger wave-V amplitude for chirp than for click stimulation in all masking conditions. Wave-V amplitude for the chirp increased continuously with increasing high-pass cutoff frequency while it remains nearly constant for the click for cutoff frequencies greater than 1 kHz. The same two stimuli were tested in the presence of a notched-noise masker with one-octave wide spectral notches corresponding to the cutoff frequencies used in the first experiment. The recordings were compared with derived responses, calculated offline, from the high-pass masking conditions. No significant difference in response amplitude between click and chirp stimulation was found for the notched-noise responses as well as for the derived responses. In the second experiment, responses were obtained using narrow-band stimuli. A low-frequency chirp and a 250-Hz tone pulse with comparable duration and magnitude spectrum were used as stimuli. The narrow band chirp elicited a larger response amplitude than the tone pulse at low and medium stimulation levels. Overall, the results of the present study further demonstrate the importance of considering peripheral processing for the formation of ABR. The chirp might be of particular interest for assessing low-frequency information. PMID- 11931310 TI - Auditory filter nonlinearity in mild/moderate hearing impairment. AB - Sensorineural hearing loss has frequently been shown to result in a loss of frequency selectivity. Less is known about its effects on the level dependence of selectivity that is so prominent a feature of normal hearing. The aim of the present study is to characterize such changes in nonlinearity as manifested in the auditory filter shapes of listeners with mild/moderate hearing impairment. Notched-noise masked thresholds at 2 kHz were measured over a range of stimulus levels in hearing-impaired listeners with losses of 20-50 dB. Growth-of-masking functions for different notch widths are more parallel for hearing-impaired than for normal-hearing listeners, indicating a more linear filter. Level-dependent filter shapes estimated from the data show relatively little change in shape across level. The loss of nonlinearity is also evident in the input/output functions derived from the fitted filter shapes. Reductions in nonlinearity are clearly evident even in a listener with only 20-dB hearing loss. PMID- 11931311 TI - Auditory stream segregation on the basis of amplitude-modulation rate. AB - In this study, auditory stream segregation based on differences in the rate of envelope fluctuations--in the absence of spectral and temporal fine structure cues--was tested. The temporal sequences to segregate were composed of fully amplitude-modulated (AM) bursts of broadband noises A and B. All sequences were built by the reiteration of a ABA triplet where A modulation rate was fixed at 100 Hz and B modulation rate was variable. The first experiment was devoted to measuring the threshold difference in AM rate leading subjects to perceive the sequence as two streams as opposed to just one. The results of this first experiment revealed that subjects generally perceived the sequences as a single perceptual stream when the difference in AM rate between the A and B noises was smaller than 0.75 oct, and as two streams when the difference was larger than about 1.00 oct. These streaming thresholds were found to be substantially larger than, and not related to, the subjects' modulation-rate discrimination thresholds. The results of a second experiment demonstrated that AM-rate-based streaming was adversely affected by decreases in AM depth, but that segregation remained possible as long as the AM of either the A or B noises was above the subject's AM-detection threshold. The results of a third experiment indicated that AM-rate-based streaming effects were still observed when the modulations applied to the A and B noises were set individually, either at a constant level in dB above AM-detection threshold, or at levels at which they were of the same perceived strength. This finding suggests that AM-rate-based streaming is not necessarily mediated by perceived differences in AM depth. Altogether, the results of this study indicate that sequential sounds can be segregated on the sole basis of differences in the rate of their temporal fluctuations in the absence of other temporal or spectral cues. PMID- 11931312 TI - Spectral loudness summation as a function of duration. AB - Loudness was measured as a function of signal bandwidth for 10-, 100-, and 1000 ms-long signals. The test and reference signals were bandpass-filtered noise spectrally centered at 2 kHz. The bandwidth of the test signal was varied from 200 to 6400 Hz. The reference signal had a bandwidth of 3200 Hz. The reference levels were 45, 55, and 65 dB SPL. The level to produce equal loudness was measured with an adaptive, two-interval, two-alternative forced-choice procedure. A loudness matching procedure was used, where the tracks for all signal pairs to be compared were interleaved. Mean results for nine normal-hearing subjects showed that the magnitude of spectral loudness summation depends on signal duration. For all reference levels, a 6- to 8-dB larger level difference between equally loud signals with the smallest (delta f = 200 Hz) and largest (delta f = 6400 Hz) bandwidth is found for 10-ms-long signals than for the 1000-ms-long signals. The duration effect slightly decreases with increasing reference loudness. As a consequence, loudness models should include a duration-dependent compression stage. Alternatively, if a fixed loudness ratio between signals of different duration is assumed, this loudness ratio should depend on the signal spectrum. PMID- 11931313 TI - Informational masking with small set sizes. AB - Informational masking refers to interference in the detectability of a sound, or discrimination of some property of a sound, beyond that which can be attributed to interactions at the auditory periphery. In the current experiments the signal to be detected was a tone added to a 6-tone masker, and informational masking was introduced by randomly choosing the frequencies of the tones that comprise the masker. The primary question was whether small numbers of maskers could replace randomly drawn maskers without sacrificing the underlying detection schemes adopted by observers. Similar to the method used by Wright and Saberi [J. Acoust. Soc. Am. 105, 1765-1775 (1999)], detection thresholds were measured for different masker set sizes, where set size refers to the number of 6-tone maskers from which any one masker was drawn. Set sizes of 3, 6, 12, and 24 were tested as well as conditions in which the maskers were chosen at random. In addition, observers' memory for maskers was coarsely evaluated. Large differences in thresholds were found across observers and across different masker sets. Even for set sizes of 24, the memory test suggests some recognition of maskers for some observers. Post hoc analysis of the data included an evaluation of the relative contribution of different frequencies using a single linear model. As a base for comparison, a linear model fitted to each condition was also evaluated. Although the data were fitted better using many rather than one linear model, the reduction in quality of fit was modest. This result suggests substantial consistency in decision strategies regardless of masker set size. PMID- 11931314 TI - Similarity, uncertainty, and masking in the identification of nonspeech auditory patterns. AB - This study examined whether increasing the similarity between informational maskers and signals would increase the amount of masking obtained in a nonspeech pattern identification task. The signals were contiguous sequences of pure-tone bursts arranged in six narrow-band spectro-temporal patterns. The informational maskers were sequences of multitone bursts played synchronously with the signal tones. The listener's task was to identify the patterns in a 1-interval 6 alternative forced-choice procedure. Three types of multitone maskers were generated according to different randomization rules. For the least signal-like informational masker, the components in each multitone burst were chosen at random within the frequency range of 200-6500 Hz, excluding a "protected region" around the signal frequencies. For the intermediate masker, the frequency components in the first burst were chosen quasirandomly, but the components in successive bursts were constrained to fall in narrow frequency bands around the frequencies of the components in the initial burst. Within the narrow bands the frequencies were randomized. This masker was considered to be more similar to the signal patterns because it consisted of a set of narrow-band sequences any one of which might be mistaken for a signal pattern. The most signal-like masker was similar to the intermediate masker in that it consisted of a set of synchronously played narrow-band sequences, but the variation in frequency within each sequence was sinusoidal, completing roughly one period in a sequence. This masker consisted of discernible patterns but not patterns that were part of the set of signals. In addition, masking produced by Gaussian noise bursts--thought to produce primarily peripherally based "energetic masking"--was measured and compared to the informational masking results. For the three informational maskers, more masking was produced by the maskers comprised of narrow-band sequences than for the masker in which the frequencies were not constrained to narrow bands. Also, the slopes of the performance-level functions for the three informational maskers were much shallower than for the Gaussian noise masker or for no masker. The findings provided qualified support for the hypothesis that increasing the similarity between signals and maskers, or parts of the maskers, causes greater informational masking. However, it is also possible that the greater masking was a consequence of increasing the number of perceptual "streams" that had to be evaluated by the listener. PMID- 11931315 TI - Learning to perceive pitch differences. AB - This paper reports two experiments concerning the stimulus specificity of pitch discrimination learning. In experiment 1, listeners were initially trained, during ten sessions (about 11,000 trials), to discriminate a monaural pure tone of 3000 Hz from ipsilateral pure tones with slightly different frequencies. The resulting perceptual learning (improvement in discrimination thresholds) appeared to be frequency-specific since, in subsequent sessions, new learning was observed when the 3000-Hz standard tone was replaced by a standard tone of 1200 Hz, or 6500 Hz. By contrast, a subsequent presentation of the initial tones to the contralateral ear showed that the initial learning was not, or was only weakly, ear-specific. In experiment 2, training in pitch discrimination was initially provided using complex tones that consisted of harmonics 3-7 of a missing fundamental (near 100 Hz for some listeners, 500 Hz for others). Subsequently, the standard complex was replaced by a standard pure tone with a frequency which could be either equal to the standard complex's missing fundamental or remote from it. In the former case, the two standard stimuli were matched in pitch. However, this perceptual relationship did not appear to favor the transfer of learning. Therefore, the results indicated that pitch discrimination learning is, at least to some extent, timbre-specific, and cannot be viewed as a reduction of an internal noise which would affect directly the output of a neural device extracting pitch from both pure tones and complex tones including low-rank harmonics. PMID- 11931316 TI - Decision strategies of hearing-impaired listeners in spectral shape discrimination. AB - The ability to discriminate between sounds with different spectral shapes was evaluated for normal-hearing and hearing-impaired listeners. Listeners detected a 920-Hz tone added in phase to a single component of a standard consisting of the sum of five tones spaced equally on a logarithmic frequency scale ranging from 200 to 4200 Hz. An overall level randomization of 10 dB was either present or absent. In one subset of conditions, the no-perturbation conditions, the standard stimulus was the sum of equal-amplitude tones. In the perturbation conditions, the amplitudes of the components within a stimulus were randomly altered on every presentation. For both perturbation and no-perturbation conditions, thresholds for the detection of the 920-Hz tone were measured to compare sensitivity to changes in spectral shape between normal-hearing and hearing-impaired listeners. To assess whether hearing-impaired listeners relied on different regions of the spectrum to discriminate between sounds, spectral weights were estimated from the perturbed standards by correlating the listener's responses with the level differences per component across two intervals of a two-alternative forced-choice task. Results showed that hearing-impaired and normal-hearing listeners had similar sensitivity to changes in spectral shape. On average, across-frequency correlation functions also were similar for both groups of listeners, suggesting that as long as all components are audible and well separated in frequency, hearing-impaired listeners can use information across frequency as well as normal hearing listeners. Analysis of the individual data revealed, however, that normal hearing listeners may be better able to adopt optimal weighting schemes. This conclusion is only tentative, as differences in internal noise may need to be considered to interpret the results obtained from weighting studies between normal-hearing and hearing-impaired listeners. PMID- 11931317 TI - Maximum speed of pitch change and how it may relate to speech. AB - How fast speakers can change pitch voluntarily is potentially an important articulatory constraint for speech production. Previous attempts at assessing the maximum speed of pitch change have helped improve understanding of certain aspects of pitch production in speech. However, since only "response time"--time needed to complete the middle 75% of a pitch shift--was measured in previous studies, direct comparisons with speech data have been difficult. In the present study, a new experimental paradigm was adopted in which subjects produced rapid successions of pitch shifts by imitating synthesized model pitch undulation patterns. This permitted the measurement of the duration of entire pitch shifts. Native speakers of English and Mandarin participated as subjects. The speed of pitch change was measured both in terms of response time and excursion time-time needed to complete the entire pitch shift. Results show that excursion time is nearly twice as long as response time. This suggests that physiological limitation on the speed of pitch movement is greater than has been recognized. Also, it is found that the maximum speed of pitch change varies quite linearly with excursion size, and that it is different for pitch rises and falls. Comparisons of present data with data on speed of pitch change from studies of real speech found them to be largely comparable. This suggests that the maximum speed of pitch change is often approached in speech, and that the role of physiological constraints in determining the shape and alignment of F0 contours in speech is probably greater than has been appreciated. PMID- 11931318 TI - Electromagnetic articulograph based on a nonparametric representation of the magnetic field. AB - Electromagnetic articulograph (EMA) devices are capable of measuring movements of the articulatory organs inside and outside the vocal tract with fine spatial and temporal resolutions, thus providing useful articulatory data for investigating the speech production process. The position of the receiver coil is detected in the EMA device on the basis of a field function representing the spatial pattern of the magnetic field in relation to the relative positions of the transmitter and receiver coils. Therefore, the design and calibration of the field function are quite important and influence the accuracy of position detection. This paper presents a nonparametric method for representing the magnetic field, and also describes a method for determining the receiver position from the strength of the induced signal in the receiver coil. The field pattern in this method is expressed by using a multivariate spline as a function of the position in the device's coordinate system. Because of the piecewise property of the basis functions and the freedom in the selection of the rank and the number of the basis functions, the spline function has a superior ability to flexibly and accurately represent the field pattern, even when it suffers from fluctuations caused by the interference between the transmitting channels. The position of the receiver coil is determined by minimizing the difference between the measured strength of the received signal and the predicted one from the spline representation of the magnetic field. Experimental results show that the error in estimating the receiver position is less than 0.1 mm for a 14 x 14-cm measurement area, and this error can be further reduced by using a spline-smoothing technique. PMID- 11931319 TI - Intensity-importance functions for bandlimited monosyllabic words. AB - A study was carried out to determine the relative importance to speech intelligibility of different intensities within the speech dynamic range. The functions that were derived are analogous to previous descriptions of the relative importance of different frequencies and are referred to here as intensity-importance functions (IIFs). They were obtained as follows. Sharply filtered bands of speech (NU6 monosyllabic words) were mixed with filtered noise and presented alone or in pairs at 19 signal-to-noise ratios (-25 to 41 dB). When paired bands were tested, the level and signal-to-noise ratio (SNR) of one band were held constant while the level and SNR of the other band were varied. The listeners were 100 normal hearers, organized into five 20-person groups. Each group provided speech recognition data for one of five frequency regions (141 562, 562-1122, 1122-1778, 1778-2818, and 2818-8913 Hz). Comparisons of the results for each group indicated that IIFs vary with frequency and SNR. Current methods for predicting intelligibility from physical measurements of speech audibility would need to be revised in order to take such findings into consideration. PMID- 11931320 TI - Bell clapper impact dynamics and the voicing of a carillon. AB - The periodic re-voicing of the bell clappers of the Australian National Carillon in Canberra provided an opportunity for the study of the acoustic effects of this operation. After prolonged playing, the impact of the pear-shaped clapper on a bell produces a significant flat area on both the clapper and the inside surface of the bell. This deformation significantly decreases the duration of the impact event and has the effect of increasing the relative amplitude of higher modes in the bell sound, making it "brighter" or even "clangy." This effect is studied by comparing the spectral envelope of the sounds of several bells before and after voicing. Theoretical analysis shows that the clapper actually strikes the bell and remains in contact with the bell surface until it is ejected by a displacement pulse that has traveled around the complete circumference of the bell. The contact time, typically about 1 ms, is therefore much longer than the effective impact time, which is only a few tenths of a millisecond. Both the impact time and the contact time are reduced by the presence of a flat on the clapper. PMID- 11931321 TI - Ultrasonic wave speed measurement using the time-delay profile of rf backscattered signals: simulation and experimental results. AB - Conventional methods determine the ultrasonic wave speed measuring the medium path length propagated by a pulsed wave and the corresponding time-of-flight. In this work, the wave speed is determined without the need of the path length. A transmit transducer sends a pulsed wave into the medium (wave speed constant along the beam axis) and the backscattered signal is collected by a hydrophone placed at two distinct positions near the transmitted beam. The time-delay profile, between gated windows of the two rf-signals received by the hydrophone, is determined using a cross-correlation method. Also, a theoretical time-delay profile is determined considering the wave speed as a parameter. The estimated wave speed is obtained upon minimization of the rms error between theoretical and experimental time-delay profiles. A PZT conically focused transmitting transducer with center frequency of 3.3 MHz, focal depth of 30 mm, and beam full width (-3 dB) of 2 mm at the focus was used together with a PZT hydrophone (0.8 mm of aperture). The method was applied to three phantoms (wave speed of 1220, 1540, and 1720 m/s) and, in vitro, to fresh bovine liver sample, immersed in a temperature-controlled water bath. The results present a relative speed error less than 3% when compared with the sound speed obtained by a conventional method. PMID- 11931322 TI - A blood vessel exposed to ultrasound: a mathematical simulation of the temperature field. AB - In this article we present a mathematical simulation of the temperature field in and around a blood vessel when it is sonicated by a focused ultrasound beam. A simplified geometry is considered: a cylindrical blood vessel is embedded in tissue parallel to a flat skin surface. The ultrasound transducer is placed on the skin above the blood vessel, perpendicular to the skin surface. The 3D geometry of the problem is simplified by transformation, which maps the domain into a parallelepiped. A computational algorithm and computer program were developed. The simulation provides the conditions for successful occlusion of a blood vessel and demonstrates the significant role of the blood flow rate on the temperature difference between the vessel wall and the surrounding tissue. Comparing the predictions with published experimental data tested the validity of the method. PMID- 11931323 TI - Reparable sonoporation generated by microstreaming. AB - Reparable sonoporation was observed in Jurkat lymphocytes in suspension exposed to a vibrating Mason horn tuned to 21.4 KHz. The diameter of the horn tip was 400 microm and its transverse displacement amplitude was 7.8 microm. It was found that the shear stress associated with microstreaming surrounding the Mason-horn tip was the primary reason for the cell reparable sonoporation. The threshold shear stress was determined to be 12 +/- 4 Pa for exposure time up to 7 min. It was also found that the shorter the exposure time, the greater the threshold. PMID- 11931324 TI - Acoustic features of male baboon loud calls: influences of context, age, and individuality. AB - The acoustic structure of loud calls ("wahoos") recorded from free-ranging male baboons (Papio cynocephalus ursinus) in the Moremi Game Reserve, Botswana, was examined for differences between and within contexts, using calls given in response to predators (alarm wahoos), during male contests (contest wahoos), and when a male had become separated from the group (contact wahoos). Calls were recorded from adolescent, subadult, and adult males. In addition, male alarm calls were compared with those recorded from females. Despite their superficial acoustic similarity, the analysis revealed a number of significant differences between alarm, contest, and contact wahoos. Contest wahoos are given at a much higher rate, exhibit lower frequency characteristics, have a longer "hoo" duration, and a relatively louder "hoo" portion than alarm wahoos. Contact wahoos are acoustically similar to contest wahoos, but are given at a much lower rate. Both alarm and contest wahoos also exhibit significant differences among individuals. Some of the acoustic features that vary in relation to age and sex presumably reflect differences in body size, whereas others are possibly related to male stamina and endurance. The finding that calls serving markedly different functions constitute variants of the same general call type suggests that the vocal production in nonhuman primates is evolutionarily constrained. PMID- 11931325 TI - Two-tone suppression in the cricket, Eunemobius carolinus (Gryllidae, Nemobiinae). AB - Sounds with frequencies >15 kHz elicit an acoustic startle response (ASR) in flying crickets (Eunemobius carolinus). Although frequencies <15 kHz do not elicit the ASR when presented alone, when presented with ultrasound (40 kHz), low frequency stimuli suppress the ultrasound-induced startle. Thus, using methods similar to those in masking experiments, we used two-tone suppression to assay sensitivity to frequencies in the audio band. Startle suppression was tuned to frequencies near 5 kHz, the frequency range of male calling songs. Similar to equal loudness contours measured in humans, however, equal suppression contours were not parallel, as the equivalent rectangular bandwidth of suppression tuning changed with increases in ultrasound intensity. Temporal integration of suppressor stimuli was measured using nonsimultaneous presentations of 5-ms pulses of 6 and 40 kHz. We found that no suppression occurs when the suppressing tone is >2 ms after and >5 ms before the ultrasound stimulus, suggesting that stimulus overlap is a requirement for suppression. When considered together with our finding that the intensity of low-frequency stimuli required for suppression is greater than that produced by singing males, the overlap requirement suggests that two-tone suppression functions to limit the ASR to sounds containing only ultrasound and not to broadband sounds that span the audio and ultrasound range. PMID- 11931326 TI - Duchenne muscular dystrophy--parental perceptions. AB - Quality of life and availability of services are important for boys with Duchenne muscular dystrophy (DMD) and their families. Families attending our neuromuscular clinic completed a questionnaire on parental perception regarding the importance of services, health issues, and quality of life issues both "now" and "in the future." Eighty-nine percent of the families (31/35) completed questionnaires. Services and health issues related to prolonging ambulation were most important, especially for the parents of younger boys. Mental health issues such as social isolation, anger, and depression were very important, particularly for the families of older boys and were anticipated to be more important in the future. Pediatricians should be aware of both the immediate needs of families to meet the physical and emotional challenges of DMD and the increasing requirement to address the social needs of these patients and their families as the boys become older. PMID- 11931327 TI - Child care program directors' level of knowledge about asthma and factors associated with knowledge. AB - Although asthmatic children spend a significant amount of time in child care programs, little is known about child care program directors' knowledge about asthma and what factors influence directors' level of knowledge about asthma. A telephone interview was conducted with directors of a randomly selected sample of all licensed child care centers and group day care homes in Connecticut. Directors of most child care programs were willing to admit asthmatic children to their programs, but their knowledge about asthma needs to be improved. Directors who have served for long periods of time may be a target group for additional education about asthma. PMID- 11931328 TI - Severe thrombocytopenic purpura as a complication of cat scratch disease. PMID- 11931329 TI - Unmasking of absence seizures by carbamazepine. PMID- 11931330 TI - A rare cause of respiratory distress in an eleven-month-old. PMID- 11931331 TI - Tonsil tummy tumult. PMID- 11931332 TI - Acquired brachial arteriovenous fistula in an ex-premature infant. PMID- 11931333 TI - Simple noninvasive effective method for removal of nasal foreign bodies in infants and children. PMID- 11931334 TI - Focal vs. generalized seizures: why classify? PMID- 11931335 TI - Bronchopulmonary dysplasia (chronic lung disease of infancy): an update for the pediatrician. AB - This paper is a review of the changes that have occurred over the past 35 years in the clinical and radiologic presentation of bronchopulmonary dysplasia (BPD), now more commonly referred to as chronic lung disease of infancy (CLD). Curent thoughts on etiology are only briefly discussed. The major focus is the management of the patient with CLD by the primary care physician once the patient has been discharged from hospital. Oxygen, diuretic, bronchodilator, antiinflammatory and nutritional therapies are discussed in detail. Finally, current information on long-term prognosis and recommendations for the prevention of RSV and influenza infections are reviewed. PMID- 11931336 TI - Spanking. Point. PMID- 11931337 TI - Spanking. Counterpoint. PMID- 11931338 TI - Kindergarten health assessment reports: what do schools really learn from them? AB - The kindergarten health assessment report (KHAR), mandated by most states, is used to identify children at school entry with any health problems that may interfere with school performance. The objective of this study was to review the completeness and accuracy of the reports that schools receive from health care providers. By analyzing 3,952 KHARs of children enrolled in the Guilford County (North Carolina) Public Schools during the 1999-2000 school year we found that only 20% were fully completed and only 32% recorded results of all 6 required screening tests. Results of the 3 screening tests most applicable to school readiness: vision, hearing, and developmental screening, were documented only 55% of the time. Providers failed to properly classify 75% of children who were either underweight or overweight. Abnormal vision screening results were noted in 485 (14.2%) children, of whom only 38% were recommended for follow-up. Results of vision and hearing screening were recorded in only 50% of children noted to have developmental concerns. The information recorded on the kindergarten health assessment forms is incomplete and frequently inaccurate. These findings arouse concern, given that these forms constitute the basis for school districts to identify those children who may have medical problems. PMID- 11931339 TI - Early newborn discharge: a tale of two hospitals. AB - This study reviews physician documentation of compliance with The American Academy of Pediatrics (AAP) policy RE9539 regarding early newborn hospital discharge and follow-up. All pediatricians in Kalamazoo, MI, were educated at a grand rounds regarding the AAP early newborn discharge policy. Newborns are seen at 2 community hospitals. One of them simultaneously instituted a Quality Improvement/Feedback (QI/F) program regarding early newborn discharge, (intervention) and the other 1 did not (control). This is a retrospective chart analysis that compares performance of 4 pediatric practices seeing newborns at each hospital. Each practice was compared at each hospital for appropriateness of discharge orders before and after the educational grand rounds and the QI/F initiative. Statistical analysis was done using the chi square test and the Breslow-Day test for homogeneity, and the Fisher's Exact Test. Odds ratios with a 95% confidence interval based on Taylor's approximation were used. There were no significant differences between the pediatric practices' performance before and after the educational initiative at the control hospital. There were significant differences before and after the educational initiative in the intervention hospital with the QI/F initiative. There was a significant reduction in variation among the practices after the QI/F initiative at the intervention hospital. When both hospitals were compared after the educational initiative, there was a significant difference between compliance among the same practices at each hospital, with better compliance at the intervention hospital with the QI/F initiative. There were significant differences in physicians' performance at the intervention hospital before and after the educational and QI/F initiatives. However, it was noted that the very same physicians did not comply as well in the control hospital without the QI/F initiative, thus still raising questions as to whether QI measures alter physician "culture." It is possible that the driving force for change in physician behavior was more intragroup peer pressure than an external QI/F initiative. PMID- 11931340 TI - The human PER1 gene is inducible by interleukin-6. AB - The mammalian period (Per) genes which are components of the circadian clock are mainly regulated via an autoregulatory feedback loop. Here we show that a human PER1 (hPER1) reporter gene activity is stimulated by interleukin-6 (IL-6), a member of the large cytokine gene family and an inducer of the acute phase reaction, in human hepatoma (HuH-7) cells. Our results confirm and extend the view that the hPER1 promoter acts as a sensor for multiple signaling molecules thereby integrating different physiological parameters. PMID- 11931341 TI - Routing of membrane proteins to large dense core vesicles in PC12 cells. AB - Routing of membrane proteins to large dense core vesicles in neuroendocrine cells can depend on information in both the lumenal and cytoplasmic domains. This study in PC12 cells focuses on the routing, cleavage, and secretion of an integral membrane protein, peptidylglycine alpha-amidating monooxygenase (PAM), examining both endogenous and virally derived membrane PAM. The role of the lumenal catalytic domains in membrane PAM trafficking was examined by replacement with an epitope tag. Virally derived membrane PAM is localized to the perinuclear area and to slender processes where the large dense core vesicles are located. Expression of PAM along with a neuroendocrine-specific endoprotease liberates a soluble monooxygenase domain, yielding regulated secretion of both the monooxygenase and the prohormone convertase from large dense core vesicles. The subcellular distribution of the epitope-substituted version of PAM within the cells is similar to that of membrane PAM, and both proteins are internalized from the plasma membrane. When secretion is stimulated, Serine937 in the cytoplasmic domain of PAM is phosphorylated to a similar extent in endogenous membrane PAM, virally encoded membrane PAM, and epitope-substituted PAM. Thus, the lumenal PAM catalytic domains are not required for routing or phosphorylation of PAM in PC12 cells. PMID- 11931342 TI - Neutral endopeptidase knockout induces hyperalgesia in a model of visceral pain, an effect related to bradykinin and nitric oxide. AB - Neutral endopeptidase (EC3.4.24.11, NEP, enkephalinase) is a zinc metalloendopeptidase, cleaving a variety of substrates like enkephalins, substance P, and bradykinin. In the brain, NEP is a key enzyme in the degradation of enkephalins. Pharmacological inhibition of NEP-activity causes analgesia resulting from enhanced extracellular enkephalin concentrations. Recently, transgenic mice lacking the enzyme NEP have been developed (Lu, 1995). The present study was designed to investigate the nociceptive behavior of these NEP knockout mice. Interestingly, NEP-deficient mice did not respond with decreased pain perception, but exhibited hyperalgesia in the hot-plate jump, warm-water tail-withdrawal, and mostnotablyin theacetic-acid writhing test. Inhibition of aminopeptidase N by bestatin reduced writhing in both strains, whereas NEP inhibition by thiorphan reduced writhing selectively in wild-type mice. Naloxone increased writhing in wild-type but not in knockouts, whereas the bradykinin B2 receptor antagonist HOE140 reduced writhing selectively in NEP-knockouts. Similarly, the nitric oxide synthase inhibitor L-NAME reduced writhing in NEP knockouts. These results indicate that genetic elimination of NEP, in contrast to pharmacological inhibition, leads to bradykinin-induced hyperalgesia instead of enkephalin-mediated analgesia. Nitric oxide (NO) is suggested to be involved in this process. PMID- 11931343 TI - Reinforcing effect of subcutaneous morphine in a modified Ettenberg runway. AB - Alley running has been successfully used as an operant to demonstrate both the positive and negative reinforcing effect of intravenously administered drugs of abuse in a bona fide operant conditioning paradigm, the Ettenberg runway, in which confounding drug effects on motor performance and drug accumulation are avoided. While Ettenberg and colleagues focus on the intravenous route of drug administration, we tested the practicability of the subcutaneous route of administration in this runway paradigm in Sprague Dawley rats, using morphine as the investigated drug of abuse. We also modified the Ettenberg runway, most notably in that either food (sweetened condensed milk), no food, morphine, or saline was presented outside the runway in a separate cage. This made shaping, i.e., the initial presentation of a food reinforcer within the runway, necessary to establish responding. The manipulations necessary to administer subcutaneous (sc) injections were well tolerated by over 90% of the tested rats (n = 93). However, sc injections increased runtimes to the experimenter cutoff of 60 s within 20 once-daily sessions. Because of strong experimenter effects, all morphine doses or saline had to be adminstered blind. Under these experimenter blind conditions, 0.1 and 1 mg/kg subcutaneous morphine proved to be reinforcing in that these doses significantly slowed down the gradual increase in runtimes imposed upon by the sc injection procedure. Thus, morphine can be demonstrated to be a positive reinforcer in a modified Ettenberg runway even when given subcutaneously. This effect, however, is eventually overcome by the negative reinforcing effect of subjecting the animals to sc injection procedure. PMID- 11931344 TI - Venlafaxine and mirtazapine: different mechanisms of antidepressant action, common opioid-mediated antinociceptive effects--a possible opioid involvement in severe depression? AB - The efficacy of each antidepressant available has been found equal to that of amitriptyline in double-blind studies as far as mild to moderate depression is involved. However, it seems that some antidepressants are more effective than others in the treatment of severe types of depression (i.e., delusional depression and refractory depression). Following studies regarding the antinociceptive mechanisms of various antidepressants, we speculate that the involvement of the opioid system in the antidepressants' mechanism of action may be necessary, in order to prove effective in the treatment of severe depression. Among the antidepressants of the newer generations, that involvement occurs only with venlafaxine (a presynaptic drug which blocks the synaptosomal uptake of noradrenaline and serotonin and, to a lesser degree, of dopamine) and with mirtazapine (a postsynaptic drug which enhances noradrenergic and 5-HT1A-mediated serotonergic neurotransmission via antagonism of central alpha-auto- and hetero adrenoreceptors). When mice were tested with a hotplate analgesia meter, both venlafaxine and mirtazapine induced a dose-dependent, naloxone-reversible antinociceptive effect following ip administration. Summing up the various interactions of venlafaxine and mirtazapine with opioid, noradrenergic and serotonergic agonists and antagonists, we found that the antinociceptive effect of venlafaxine is influenced by opioid receptor subtypes (mu-, kappa1- kappa3- and delta-opioid receptor subtypes) combined with the alpha2-adrenergic receptor, whereas the antinociceptive effect of mirtazapine mainly involves mu- and kappa3 opioid mechanisms. This opioid profile of the two drugs may be one of the explanations to their efficacy in severe depression, unlike the SSRIs and other antidepressants which lack opioid activity. PMID- 11931345 TI - Drug design at peptide receptors: somatostatin receptor ligands. AB - Somatostatin (SRIF, somatotropin release inhibiting factor), discovered for its inhibitory action on growth hormone (GH) secretion from pituitary, is an abundant neuropeptide. Two forms, SRIF14 and SRIF28 exist. Recently, a second family of peptides with very similar sequences and features was described; the cortistatins (CST), CST17 and CST29 which are brain selective. The five cloned SRIF receptors (sst1-5) belong to the G-protein coupled/ heptathelical receptor family. Structural and operational features distinguish two classes of receptors; SRIF1 - sst2/sst3/sst5 (high affinity for octreotide or seglitide) and SRIF2 = sst1/sst4(very low affinitty for the aforementioned ligands). The affinity of SRIF receptors for somatostatins and cortistatins is equally high, and it is not clear whether selective receptors do exist for one or the other of the peptides. Several radiologlands label all SRIF receptors, e.g., [125]LTT-SRIF28' [l25I]CGP23996, [125]Tyr10cortistatin or [125I]Tyr11SRIF14. In contrast, [125I]Tyr3octreotide, [125I]BIM23027, [125I]MK678 or [125I]D-Trp8SRIF14 label predominantly SRIF1 sites, especially sst2 and possibly sst5 receptors. In brain, [125I]Tyr3octreotide binding equates with sst2 receptor mRNA distribution. Native SRIF2receptors can be labeled with [125I]SRIF14 in the presence of high NaCl in brain (sst1) or lung (sst4) tissue. Short cyclic or linear peptide analogs show selectivity for sst2/sst5 (octreotide, lanreotide, BIM 23027), sst1 (CH-275), sst3 (sst3-ODN-8), or sst5 receptors (BIM 23268); although claims for selectivity have not always been confirmed. Beta peptides ith affinity for SRIF receptors are also reported. The general lack of SRIF receptor antagonists is unique for peptide receptors, although CYN 154806 is a selective and potent sst2 antagonist. Nonpeptide ligands are still rare, although a number of molecules have been reported with selectivity and potency for sst1 (L 757,519), sst2 (L 779,976), sst3 (L 796,778), sst4 (NNC 26-9100, L 803,087) or sst1/sst5 receptors (L 817,018). Such molecules are essential to establish the role of SRIF receptors, e.g., sst1 in hypothalamic glutamate currents: sst2 in inhibiting release of GH, glucagon, TSH, gastric acid secretion, pain, seizures and tumor growth, and sst5 in vascular remodeling and inhibition of insulin and GH release. PMID- 11931346 TI - Evaluation of an anxiety-related phenotype in galanin overexpressing transgenic mice. AB - Understanding the role of neuropeptides in mediating emotional behaviors is an important avenue for discovering novel drug targets for anxiety disorders. A role for galanin in mediating anxiety-related behavior is suggested by the pattern of distribution in the CNS and the coexistence of galanin with norepinephrine in the locus coeruleus. Studies in rats have shown that central administration of galanin modulates anxiety-related behaviors, and galanin release blocks the proanxiety effects of noradrenergic activation in prestressed rats. To further investigate the role of galanin in anxiety behaviors, we conducted a comprehensive behavioral phenotyping of galanin overexpressing transgenic mice (GAL-tg). GAL-tg mice were normal on measures of general health, neurological reflexes, home cage social behaviors, sensory functions, motor coordination, and exploratory locomotor activity. In three separate tests for anxiety-related behaviors, the elevated plus-maze, light <--> dark exploration, and open field center time, GAL-tg mice showed no anxiety-like phenotype. GAL-tg mice and wild type littermate controls were equally responsive to the anxiolytic effects of chlordiazepoxide (10 mg/kg) in the light <--> dark exploration test, indicating normal benzodiazepine receptor function in GAL-tg mice. Stimulation of noradrenergic cells via administration with an alpha2 adrenoreceptor antagonist, yohimbine (2.5 mg/kg), produced proanxiety effects in wild type mice in the light <--> dark exploration test, but not in the GAL-tg mice. These data suggest that galanin contributes to the modulation of anxiety states induced by high levels of noradrenergic activation, but is silent under less challenging situations. A specific role for galanin in extreme anxiety states represents an attractive target for the development of novel anxiolytic treatments. PMID- 11931347 TI - (N-stearyl, norleucine17)VIPhybrid is a broad spectrum vasoactive intestinal peptide receptor antagonist. AB - The effects of a (N-stearyl, Norleucine17) vasoactive intestinal peptide hybrid ((SN)VIPhybrid) on cells stably transfected with VPAC,, VPAC2, or PAC1 receptors were investigated. (SN)VIPhybrid inhibited specific 125I-VIP binding to membranes derived from CHO cells transfected with VPAC, or VPAC2 receptors with high affinity (IC50 = 30 and 50 nM). (SN)VIPhyb inhibited specific 125I-PACAP-27 binding to membranes derived from NIH/3T3 cells transfected with PAC1 receptors with high affinity (IC50 = 65 nM). PACAP-27 caused cAMP elevation in NIH/3T3 cells transfected with PAC1 receptors and the increase cAMP caused by pituitary adenylated cyclase (PACAP) was inhibited by (SN)VIPhyb. Also, the increase in cAMP caused by VIP using CHO cells transfected with VPAC1 or VPAC2 receptors was antagonized by (SN)VIPhyb. These results indicate that (SN)VIPhyb is an antagonist for VPAC1, VPAC2, and PAC1 receptors. PMID- 11931348 TI - A single administration of the peptide NAP induces long-term protective changes against the consequences of head injury: gene Atlas array analysis. AB - The femtomolar-acting eight-amino-acid peptide (NAP), derived from activity dependent neuroprotective protein (ADNP), provides long-term protection against the deleterious effects of closed head injury (CHI) in mice. Fifteen minutes after injury, mice were divided into two groups, control and NAP-treated and a single subcutaneous injection of NAP or vehicle was administered. A third group served as sham-treated (not subjected to head trauma). Each mouse was assessed for its clinical function, using neurological severity score, at various time intervals following CHI, up to 30-45 d. Total cerebral cortex RNA was prepared from the site of injury of CHI mice, and from parallel regions in peptide-treated and sham brains. RNA was then reversed transcribed to yield radioactive cDNA preparations that were hybridized to Atlas array membranes containing 1200 cDNAs spots. Comparison of sham-treated individual mice showed differential expression levels of at least 15 mRNA species. Furthermore, results indicated that one of the genes that did not change among individuals but specifically increased after CHI and decreased after NAP treatment was the cell surface glycoprotein Mac-1 (CD11B antigen). Thus, Mac-1 is suggested as a marker for the long-term outcome of head injury and as a potential target for NAP protective actions. PMID- 11931349 TI - Pituitary adenylate cyclase-activating polypeptide and vasoactive intestinal peptide-stimulated cyclic AMP synthesis in rat cerebral cortical slices: interaction with noradrenaline, adrenaline, and forskolin. AB - Pituitary adenylate cyclase-activating polypeptide (PACAP; 0.001-1 microM) and vasoactive intestinal peptide (VIP; 0.01-1 microM) produced a concentration dependent stimulation of cyclic AMP (cAMP) formation in rat cerebral cortical slices prelabeled with [3H]adenine. The effects of PACAP38 and PACAP27 were similar, and more efficacious (at 0.1 and 1 microM) than those of VIP. Adrenaline and noradrenaline (each at 100 microM) also stimulated cAMP formation, with the latter compound being more effective. Combination of PACAP38, PACAP27 (each at 0.1 microM) and VIP (1 microM) with adrenaline or noradrenaline resulted in most cases in additive effects, with some supraadditive (PACAP27 plus adrenaline) or subadditive (PACAP38 or VIP plus noradrenaline) fluctuations. In contrast, combination of each of the three peptides with 3 microM forskolin resulted in synergistic effects. These results indicate that in rat cerebral cortex there is no synergism between PACAP or VIP with noradrenaline or adrenaline; however, based on the forskolin data, it seems likely that synergistic effects may take place with VIP or PACAP and other cAMP-stimulating neuroregulators. PMID- 11931350 TI - Synaptic proteins in Alzheimer's disease. AB - Chromogranin A, chromogranin B, and secretogranin II are acidic proteins which are stored in large dense core vesicles of neurons. An antiserum, raised against a synthetic peptide (PE-11), present in the chromogranin B molecule, and an antiserum raised against secretoneurin contained in the secretogranin II sequence, was used to localize these peptides together with chromogranin A in the human hippocampal formation. The distribution of these peptides was investigated in Alzheimer's disease and compared to control subjects. Chromogranin A, chromogranin B, and secretogranin II are distinctly distributed with an overlap in their distribution patterns. They were only detected in neuronal structures. The highest density of immunoreactivity was found for chromogranin B. A layer specific distribution was especially obvious in the inner molecular layer of the dentate gyrus as secretoneurin-like immunoreactivity was restricted to its innermost part whereas that of chromogranin B was highly concentrated throughout the inner molecular layer. In Alzheimer's disease, about 10 to 20% of the amyloid immunoreactive plaques contained either chromogranin A, chromogranin B or secretoneurin. The density of secretoneurin-and chromogranin B-like immunoreactivity was significantly reduced in the inner molecular layer of the dentate gyyrs, the CA1 area, the subiculum and in layers I, III and V of the entorhinal cortex. The present study demonstrates that chromogranin peptides are markers for human hippocampal pathways. Thee are particularly suitable to study nerve fibers, terminating at the inner molecular layer of the dentate gyrus. Chromogranin peptides have a potential as neuronal markers for synaptic degeneration in Alzheimer's disease. PMID- 11931351 TI - The transcription of the hGnRH-I and hGnRH-II genes in human neuronal cells is differentially regulated by estrogen. AB - Gonadotropin releasing hormone-I (GnRH-I), a decapeptide serves as a key regulator of reproduction. Recently, several groups have identified in the mammalian brain a second form of GnRH, of unknown function, designated GnRH-II. The human neuronal medulloblastoma cells (TE-671) were recently demonstrated to express the two forms of GnRH (GnRH-I and GnRH-II). We used this cell line, as a model system, to investigate the regulation of human GnRH-I and GnRH-II genes by estrogen. Estrogen is one of the principal regulators of GnRH-I in hypothalamic neurons, acting as a classic homeostatic feedback molecule between the gonads and the brain. In this study, we investigated the regulation of the two GnRH forms by estrogen, in the human neuronal cell line TE-671. We demonstrate, for the first time, that the hGnRH-II and hGnRH-I genes are differentially regulated by estrogen. Using reverse transcriptase-polymerase chain reaction (RT-PCR) and Southern hybridization, we found that estrogen increases endogenous hGnRH-II mRNA levels and decreases endogenous hGnRH-I mRNA levels. Furthermore, we found these effects to be promoter-mediated. We cloned the hGnRH-I and hGnRH-II promoter constructs upstream to a luciferase reporter plasmid, and cotransfected these constructs with an estrogen receptor alpha into the TE-671 neuronal cells. Luciferase activity of GnRH promoter constructs treated with estrogen demonstrates that the differential regulation of the GnRH genes by estrogen is mediated at the transcription level. PMID- 11931352 TI - Interactions of glucagon-like peptide-1 (GLP-1) with the blood-brain barrier. AB - Glucagon-like peptide-1 (GLP-1) reduces insulin requirement in diabetes mellitus and promotes satiety. GLP-1 in the periphery (outside the CNS) has been shown to act on the brain to reduce food ingestion. As GLP-1 is readily degraded in blood, we focused on the interactions of [Ser8]GLP-1, an analog with similar biological effects and greater stability, with the blood-brain barrier (BBB). The influx of radiolabeled [Ser8]GLP-1 into brain has several distinctive characteristics: 1. A rapid influx rate of 8.867 +/- 0.798 x 10(4) mL/g-min as measured by multiple time regression analysis after iv injection in mice. 2. Lack of self-inhibition by excess doses of the unlabeled [Ser8]GLP-1 either iv or by in situ brain perfusion, indicating the absence of a saturable transport system at the BBB. 3. Lack of modulation by short-term fasting and some other ingestive peptides that may interact with GLP-1, including leptin, glucagon, insulin, neuropeptide Y, and melanin-concentrating hormone. 4. No inhibition of influx by the selective GLP-1 receptor antagonist exendin(9-39), suggesting that the GLP-1 receptor is not involved in the rapid entry into brain. Similarly, there was no efflux system for [Ser8]GLP-1 to exit the brain other than following the reabsorption of cerebrospinal fluid (CSF). The fast influx was not associated with high lipid solubility. Upon reaching the brain compartment, substantial amounts of [Ser8]GLP 1 entered the brain parenchyma, but a large proportion was loosely associated with the vasculature at the BBB. Finally, the influx rate of [Ser8]GLP-1 was compared with that of GLP-1 in a blood-free brain perfusion system; radiolabeled GLP-1 had a more rapid influx than its analog and neither peptide showed the self inhibition indicative of a saturable transport system. Therefore, we conclude that [Ser8]GLP-1 and the endogenous peptide GLP-1 can gain access to the brain from the periphery by simple diffusion and thus contribute to the regulation of feeding. PMID- 11931353 TI - Extrapituitary TSH in early chick embryos: Pit-1 dependence? AB - The expression of the thyrotropin (TSH) gene in the pituitary gland is thought to be dependent upon the pituitary-specific transcription factor, Pit-1. TSH immunoreactivity is, however, widespread in extrapituitary tissues, and the possibility that this may reflect a widespread distribution of Pit-1 was therefore investigated in embryonic chicks, prior to the ontogenic differentiation of the pituitary gland. TSH immunoreactivity in chick embryos at the end of the first trimester of the 21d incubation period was present in discrete cells in the developing brain (particularly in ependymal cells lining the diocoele and mesocoele and in cells lining the otic vesicle), spinal cord (ependymal cells), liver (hepatocytes), lungs (in the linings of the bronchi), gut (in the linings of the proventriculus) and limb bud (in skin, muscle, bone and nerve fibers). In some of these tissues (particularly in brain and spinal cord ependymal cells, cells in the otic vesicle and in liver hepatocytes), the distribution of TSH immunoreactivity was overlapped by the distribution of immunoreactive Pit-1, suggesting Pit-1 involvement in TSH expression in these sites. However, in other tissues (e.g., the trigeminal nerve in the head and the marginal mantle layer of the spinal cord), Pit-1 immunoreactivity was intense but TSH immunoreactivity was marginal. Conversely, other tissues (e.g., cells in the skin, blood vessels, limb bud, bronchus, proventriculus, and cardiopleural cavities) had intense TSH staining but little, if any, Pit-1 immunoreactivity. The expression of the TSH gene in these tissues would thus appear to be Pit-1 independent. These results demonstrate the presence of Pit-1 in pituitary and extrapituitary tissues of the domes tic fowl and suggest the involvement of Pit-1 in the extrapituitary expression of TSH in chick embryos may be tissue-specific. PMID- 11931356 TI - On a therapist serving as a witness. PMID- 11931354 TI - Growth hormone (GH) action in the brain: neural expression of a GH-response gene. AB - The presence of growth hormone (GH) binding sites and GH-receptor (GHR) immunoreactive proteins in the brain suggests it is a target site for GH action. This could, however, reflect the presence of GH-binding proteins (GHBP) that are not linked to intracellular signal-transduction mechanisms, rather than authentic receptors. The possibility that GH has actions in the brain therefore has been examined by determining an intracellular mediator of GH action. The mechanism of GH action involves the induction of a number of specific GH-response genes. In chickens, a novel GH-responsive gene (GHRG-1) has been identified as an intracellular marker of GH action, since this gene is not expressed in GH resistant dwarfs with dysfunctional GHRs and in normal chickens it is upregulated by exogenous GH. In normal chickens GHRG-1 mRNA is also abundant and widespread in the brain. In the cerebellum it is specifically localized in the cerebellar folia. It is present in most cells in the granular layers of the gray matter but is present in only a small number of scattered cells in the molecular layer and white matter. Intense labeling for GHRG-1 mRNA is also present in the large Purkinje cells and their dendrites at the interface between the molecular and granular layers. Labeling is also seen in the interneuronal basket cells projecting onto the Purkinje cells. In the mid-brain, cells in the ocular nerve complex and the tractus isthmo-opticus were strongly stained for GHRG-1 mRNA, with less intense staining in the central gray. In the hypothalamus, numerous small cells in periventricular locations and ependymal cells lining the III ventricle also label for GHRG-1 mRNA. These results clearly show, for the first time, the expression of a GH-responsive gene in neural tissues. Moreover, as GH- and GHR-immunoreactivity previously has been shown to be present in the same tissues expressing GHRG-1, it is possible that GH acts as an autocrine or paracrine within the CNS. PMID- 11931357 TI - Psychiatrists on the side of the angels: the Falun Gong and Soviet Jewry. PMID- 11931355 TI - Puerperal blockade of cholecystokinin (CCK1) receptors disrupts maternal behavior in lactating rats. AB - Blockade of cholecystokinin (CCK) receptors potentiates the morphine-induced disruption of maternal behavior. The present study was undertaken to determine whether treatment with lorglumide, a CCK1 antagonist during late pregnancy and early lactation can influence the maternal behavior during lactation. A possible influence of this treatment on general activity was also assessed. Twenty-seven female Wistar rats were pretreated with lorglumide (1.0mg/kg/day; sc) or saline for seven days, starting on the 17th d of pregnancy. After the withdrawal of this treatment, animals were acutely challenged with saline on day 5 and with morphine sulfate (3.0mg/kg; sc) on days 6,10, and 17 of lactation. Groups were pretreated with saline and challenged with saline (group SS) and morphine (group SM), pretreated with lorglumide and challenged with saline (group LS) and morphine (group LM). Animals were also tested for general activity on days 25 and 33 postpartum after an acute challenge with saline and morphine, respectively. Maternal behavior testing began 30 min after the acute injections at which time pups were placed throughout each mother's cage. Latencies for pup retrieval, grouping, crouching and for full maternal behavior responses were scored. Lorglumide pretreatment inhibited maternal behavior of LS vs SS group and potentiated the morphine-induced disruption of this behavior in all days of test (LM vs SM group). No significant differences were found in general activity on days 25 and 33 postpartum. These data suggest that blockade of CCK1 receptors during puerperal period has long-term implications for maternal behavior. PMID- 11931358 TI - The political misuse of psychiatry: an African-Caribbean perspective. PMID- 11931360 TI - Psychiatric abuse of Falun Gong practitioners in China. PMID- 11931359 TI - Psychiatry in its political and professional contexts: a response to Robin Munro. PMID- 11931361 TI - Comparing Soviet and Chinese political psychiatry. PMID- 11931362 TI - Political abuse of psychiatry in the Soviet Union and in China: complexities and controversies. PMID- 11931363 TI - Political abuse of psychiatry in the Soviet Union and China: a rough guide for bystanders. PMID- 11931364 TI - Code of medical ethics. PMID- 11931365 TI - Attempts to understand better the lives of violent individuals. PMID- 11931366 TI - Class action litigation in correctional psychiatry. AB - Class action litigation has been instrumental in jail and prison reform during the past two decades. Correctional mental health systems have significantly benefited from such litigation. Forensic psychiatrists have been crucial in the litigation process and the subsequent evolution of correctional mental health care systems. This article summarizes information concerning basic demographics of correctional populations and costs of correctional health care and provides a brief history of such litigation. The role of psychiatric experts, with particular reference to standards of care, is described. Specifically discussed are issues relevant to suicide prevention, the prevalence of mentally ill inmates in supermax prisons, and discharge planning. PMID- 11931367 TI - The origins of commitment for substance abuse in the United States. AB - Policymakers in the United States have long been perplexed by how to deal with substance abuse. As attitudes shifted in the 19th century toward viewing substance abuse as a medical problem akin to insanity rather than as a moral failing, greater emphasis was given to the potential for treatment. Thus, by the middle of the 19th century, states began developing substance abuse commitment codes and institutions to which substance abusers could be committed. Public ambivalence over whether substance abusers should be seen as having an illness or a weakness of will, however, was reflected in the lack of sustained support for these efforts, in contrast to support accorded systems for commitment of the mentally ill. Contemporary policymakers are faced with the same ambivalence, as they struggle with the extent to which substance abusers ought to be subjected to involuntary treatment. The legacy of the early years of substance abuse commitment lives on. PMID- 11931368 TI - The roles of behavioral health professionals in class action litigation. AB - Class action suits frequently have been used as a strategy for improving the quality of mental health care. Psychiatrists, psychologists, and professionals from related disciplines have been involved in these suits in different roles. This article presents and discusses case examples of these roles, which include the expert witness, court-appointed expert, consulting expert, monitor, special master, receiver, advocate, amicus curiae, plaintiff, and defendant. The authors caution against assuming dual roles and argue that professionals, before beginning to participate in this complex arena, should clarify their functional responsibilities, the legal basis of their involvement, and the ethics principles that will guide their actions. PMID- 11931369 TI - A British psychiatrist objects to the dangerous and severe personality disorder proposals. PMID- 11931370 TI - Countering countertransference: a forensic trainee's dilemma. AB - Forensic psychiatry involves an adaptation to a role that is very different from the psychiatrist's previous clinical experiences. To render an unbiased forensic opinion, psychiatrists have to rise above their countertransference feelings. This takes years of practice and experience. The following is an account of a forensic trainee who faced several countertransference problems as he evaluated a patient charged with attempted murder. The case report is interspersed with a review of the relevant literature. PMID- 11931371 TI - A comparison of youth referred to psychiatric emergency services: police versus other sources. AB - Although the presentation of children and adolescents to psychiatric emergency services has been increasing, little is known about the characteristics of these persons, the circumstances surrounding their referral for treatment, or their disposition. This study compares patients referred by police with those referred by others, based on data from a study of 1,779 patients in the Bronx, New York. Logistic regression was used to develop propensity scores for selection of a matched sample of non-police referrals (n = 159) to compare with police referrals (n = 53). Caregivers in police-referred cases were rated as less capable of active treatment involvement, and domestic violence was more likely to occur in their homes. Police referrals had higher substance use in the past month than referrals from other sources, were rated as more symptomatic and dangerous to self and others, had exhibited more assaultive and destructive behavior, and were less likely to be referred to outpatient services. PMID- 11931372 TI - Forensic psychiatry and the internet: practical perspectives on sexual predators and obsessional harassers in cyberspace. AB - The growth of the Internet has revolutionized how society conducts business in many areas. Not to be left behind, the sexual predator and the obsessional harasser have found cyberspace to be a vehicle capable of meeting their needs: obtaining information, monitoring and contacting victims, developing fantasy, overcoming inhibitions, avoiding apprehension, and communicating with other offenders. Although clearly disparate offender categories, these two groups are included in this article because of the likelihood of their using the Internet in pursuit of their goals. Forensic psychiatrists should become familiar with computers and the Internet so that they can conduct relevant psychiatric evaluations of such individuals and can advise attorneys, victims, and law enforcement personnel competently, when retained in that capacity. This article discusses the Internet and its use by the sexual predator and the obsessional harasser, highlighting information of interest to the forensic psychiatrist, including the poorly understood field of digital evidence. Aspects of the Internet, such as on-line dating and cybersex also are covered, because they relate to sexual predation and stalking. PMID- 11931373 TI - Political psychiatry in post-Mao China and its origins in the cultural revolution. PMID- 11931374 TI - Extracorporeal fibrinogen adsorption--efficacy, selectivity and safety in healthy subjects and patients with foot ulcers. AB - The elimination of fibrinogen from plasma improves plasma viscosity and whole blood viscosity. For extracorporeal adsorption of fibrinogen the pentapeptide gly pro-arg-pro-lys was coupled to sepharose CL-4B. Columns containing 100 ml of coupled sepharose CL-4B were used to eliminate fibrinogen from the plasma of 8 healthy male subjects (mean age 27.4 +/- 4.3 years, height 180.9 +/- 8.3 cm, weight 85.1 +/- 13.6 kg). Four treatments were performed in each proband (days 1, 2, 4 and 7). Plasma fibrinogen concentration was lowered from 221.1 +/- 39.0 to 123.5 +/- 21.7 mg/dl (2275 +/- 477 ml plasma treated) by the first treatment, from 172.8 +/- 42.3 to 105.6 +/- 16.5 mg/dl (1609 +/- 761 ml) by the second, from 140.5 +/- 13.8 to 98.8 +/- 8.6 mg/dl (1224 +/- 118 ml) by the third and from 160.2 +/- 23.6 to 106.4 +/- 9.7 mg/dl (1513 +/- 521) by the fourth. Plasma viscosity was improved from 1.40 +/- 0.18 mPa s before the first treatment to 1.23 +/- 0.06 mPa s after fourth treatment, whole blood viscosity from 4.49 +/- 0.36 mPa s to 3.83 +/- 0.27 mPa s (P < 0.01). No clinical side effects and no clinically relevant change of laboratory parameters including in vitro tests on thrombocyte function were observed. Seven men and three women (48-75 years old, 9 patients suffered from diabetes mellitus, one patient from peripheral arterial occlusive disease, 5 patients were on regular hemodialysis) were treated by fibrinogen adsorption. Each column contained 135 ml of coupled sepharose CL-4B. Treatments were scheduled on day 1, 2, 4, 6, 8, 10, 13, 16, 19, 22, 25 and 28. 144 treatments with fibrinogen adsorption were performed. No clinical side effects due to the fibrinogen-adsorption procedure were observed. In these 10 patients the fibrinogen concentration before the first treatment was 473.7 +/- 183.7 mg/dl. In the first treatment session it was lowered to 241.4 +/- 125.8 mg/dl by treating 4270 +/- 1180 ml of plasma. In the following 134 treatments the pre-treatment concentration of fibrinogen was 262.6 +/- 83.4 mg/dl, the post treatment concentration was 120.6 +/- 37.2 mg/dl. The mean volume of plasma treated was 3737 +/- 1643 ml, the mean duration of a treatment session (except the first treatment) was 143.7 +/- 63.1 min. In 7 patients a mean post-treatment fibrinogen concentration of < or = 123 mg/dl was obtained, in the other patients 133, 177 and 184 mg/dl. Yet, the decrease of fibrinogen concentration was also pronounced in these 3 patients: -82%, -67%, and -73%, respectively. Accelerated wound healing was observed in 9 of the 10 patients. In conclusion, affinity chromatography using the pentapeptide gly-pro-arg-pro-lys is an effective, selective and safe procedure to lower fibrinogen concentration in plasma thereby improving blood viscosity. It could be a therapeutic option in severe blood vessel disease where drug therapy is not sufficient and invasive procedures like bypass or angioplasty cannot be applied. PMID- 11931375 TI - Prospective evaluation of platelets prepared by single and random methods during 5 days of storage: aspects related to quality and quantity. AB - Platelets, which have an important role in hemostatic mechanisms and which were prepared by single and random methods were investigated to measure quantity, and aggregation response during 5 days of storage. The aggregation response and quantitative values of platelet concentrates (PCs), 60 of which were prepared by a single donor method and 62 by a random method were investigated during the 1st, 3rd, and 5th days of storage. The single donor platelets (SDP) were obtained by using the MCS Plus apheresis device and the random donor platelets (RDP) were obtained by two-phase centrifugation in the Heraeus 8500i centrifuge device (during the first phase, platelet rich plasma was obtained then platelet concentrate was obtained from this product) and were stored at 22 degrees C on a circular agitator. In addition, pH, PO2, PCO2, glucose and lactate values were measured in order to evaluate the effects of storage. The aggregation response was measured using adenosine diphosphate (ADP), epinephrine (EPN), collagen (COLL) and ristocetin (RIST). The cell count in mm3 and the total cell count were also measured. The total cell counts and cells in mm3 of the PCs which were prepared by the single donar method on the 1st, 3rd and 5th days, were: 3.11 x 10(11), 3.09 x 10(11), 3.07 x 10(11) and 292 x 10(3), 290 x 10(3), 289 x 10(3) and of those prepared by the random method were: 5.71 x 10(10), 5.69 x 10(10), 5.66 x 10(10) and 156 x 10(3), 153 x 10(3), 151 x 10(3). The mean aggregation responses of the PCs prepared by the two methods on the 1st, 3rd and 5th days, expressed as a % were: ADP: 94.8-93.2, 81.6-78.7, 44.3-8.2; COLL: 91.7-89.6, 79.2 74.2, 29.8-11.1; EPN: 88.5-91.3, 64.2-62.7, 39.4-4.5 and RIST: 89.4-89.4, 76.5 73.6, 14.4-3.2. Other data related to platelet storage were obtained by measuring the pH, PO2, PCO2, glucose and lactate levels of the PCs. In our study, it was determined that in spite of the optimal storage conditions, the aggregation response of the PCs decreased significantly, whereas, the numerical values changed little during the storage period. PMID- 11931376 TI - The quality of MCS+ version C2 double dose platelet concentrate with leucodepletion through a continuous filtration process. AB - 182 full double donations were collected, using 7-9 cycles with an in-house guideline based on donor HCT, and platelet count. 95.6% of donations were within specifications for platelet yield and 100% were leucodepleted. To overcome the potential for red cell spill-over when the platelet peak is too small, automated continuous monitoring of machine performance has been introduced reducing the potential for spill-over. To understand the impact of the continuous exposure of platelets to the filter, the quality of the first and second part of the collection was monitored using a new set of markers for the platelet storage lesion (activation, microvesiculation, cellular injury, complement and response to ADP). The results were equivalent and compared well with routine practice. Our new protocol allows a wide range of donors to be selected and the highest percentage of products to meet the UK specification with consistency. It is recommended that it can be used for double dose platelets with routine SPM. PMID- 11931377 TI - Cytokines as quality indicators of leucoreduced red cell concentrates. AB - Different types of filters are currently used for leucodepletion of red cell concentrates. These filters meet the specification for leucoreduction (<5 x 10(6) leucocytes/ATD) but the quality of the final product may differ depending on the performance of the filters for effective removal of both leucocytes, platelets and possibly cytokines which are associated with transfusion reactions. We measured the levels of three representative cytokines: IL-8, RANTES and TGF-beta1 in red cell concentrates prior to and subsequent to the filtration procedure on day 1 and after a storage period of 35 days. Low levels of IL-8 (10-24 pg/ml) in the control unfiltered concentrates on day 1 which increased by approximately twofold on storage. Filtration reduced the levels of IL-8 on day 1 and day 35, in filtered concentrates in comparison with their control unfiltered counterparts. Leucoreduced concentrates produced by three different filters showed similar IL-8 levels on day 1 and day 35. However, concentrates prepared using another type of process showed a twofold increase in IL-8 levels on storage in comparison with day 1. None of the concentrates tested contained any detectable RANTES and TGF beta1 suggesting a minimal platelet content. These results indicate that a combination of IL-8, RANTES and TGF-beta1 are useful quality indicators for validation of leucoreduced red cell preparations. PMID- 11931378 TI - Studies on the characterisation of the cause of leucoreduction failures, with particular reference to extra gatal events. AB - The causes of leucodepletion failure are multifactoral and can be related to haematological variability in blood donors or donation, defective filters, poor specimen handling or ageing, and/or the presence of non-adhering leucocyte/platelets. Since refiltering removes all types of leucocytes, including the populations appearing as extra gated events, we have developed a practical method for refiltering the failed leucodepleted components on standard filters and back-flushing the second filter to assess the nature of the WBC sub population. In practice, recovered leucocytes from red cell filters and whole blood mainly consist of neutrophils. Those from platelet and plasma filters were mainly lymphocyte with considerable differences depending on the type of leucodepletion process. Atypical leucocytes are often seen in some pre-/post cellular leucofiltered components. These appear characteristically as small WBC with a lower affinity for filter matrix, or as cell fragment, pinched leucocyte or apoptotic cells. Different reagents in use show variable sensitivity in identifying these extra gatal events. Storage of leucodepleted samples also induces different types of abnormality in leucocyte dot plot. A useful practical approach for characterisation of the nature of leucocyte sub-populations causing failure in leucodepleted components is provided. PMID- 11931379 TI - Validation of different enrichment strategies for analysis of leucocyte sub populations: development and application of a new approach, based on leucofiltration. AB - Characterisation of the nature of leucodepletion failure, based on the analysis of leucocyte sub-populations, is an essential task for continual improvement in the clinical safety of blood components and for the selection of a safe and appropriate leucodepletion process. Such information is also critical for improving the quality of filters through an understanding of the mechanism of leucocyte removal filters. Unfortunately, the residual leucocytes of filtered blood components are around the sensitivity of current leucocyte-counting technologies, hence a need for an enrichment process. This preliminary report deals with the merits of various WBC enrichment strategies for the analysis of WBC sub-population subsets. A new procedure, based on refiltering and backflushing the content, with up to 30-fold enrichment, is described. This one step procedure has the advantage of concentrating both native and atypical leucocytes seen in blood components. The latter may account for lack of interchangeability of results obtained by various counting methods. PMID- 11931380 TI - A national quality assessment scheme for counting residual leucocytes in unfixed leucodepleted products: the effect of standardisation and 48 hour storage. AB - BACKGROUND: WBC counting, an essential part of quality monitoring of WBC-reduced blood components, is carried out logistically within 48-72 h of collection. The between-laboratory variability and effects of 24-48 h storage were investigated using three major counting technologies. STUDY DESIGN AND METHODS: Samples of RBC and platelets with WBC in the range 0-50/microl were transported by courier. WBC counting was performed on days 1 and 2, by IMAGN 2000, flow cytometry and Nageotte, initially using local protocols and then using a national flow protocol. Up to 15 laboratories participated in each exercise. RESULTS: For "real failed leucodepleted" red cell products, higher levels of variability were observed for flow and Nageotte, as compared to IMAGN. For spiked RBC samples at critical decision making point (3-20 WBC/microl), between-laboratory the coefficients of variation (CVs) were low for IMAGN and were the highest for Nageotte. Flow cytometry CVs were generally high but improved subsequent to standardisation of sampling and the gating strategy. A similar pattern in the variability of results was observed for platelet concentrates. Sign tests using all samples (carried out for each method in each exercise; 25 in total) demonstrated no overall tendency for larger WBC counts to be recorded on day 1 when compared to day 2, although this difference was significant (p < 0.001) in certain cases depending on the nature of the spiked product. CONCLUSIONS: We conclude that while a good performance is achieved using validated automated technologies for low residual leucocyte counting, the unification of reagents and standardisation of sampling and gating strategies are essential in obtaining interchangeable results. Unfixed RBC and platelet samples can generally be stored for 48 h before WBC counting. PMID- 11931381 TI - A platelet quality assessment scheme for comparing the performance of quality monitoring laboratories in the UK National Blood Service. AB - This exercise focused on performance of NBS quality monitoring establishments with respect to enumeration of low leucocyte and other quality indexes of platelet concentration. Paired identical leucodepleted platelet samples, spiked with WBC (20 cells/microl) in 'vacuette' or 'pouch' were assessed by participants (n = 20) on days 1, 2 and 5. For low WBC counting, all laboratories gave estimates within acceptable range (+/-25%) and good agreement between storage and assay methods was observed on days 1 and 2. Day 5 results showed greater variability. Under improved performance criteria (+/-15%), only one laboratory under-estimated at days 1 and 2. Similarly, other parameters demonstrated good agreement between storage methods on days 1 and 2. At day 5, mean results were often significantly different to previous days. Improved performance target (+/ 15%) will allow identification of non-conformers. PMID- 11931382 TI - Development of a microplate method for the measurement of plasma haemoglobin and its use in monitoring the storage lesion of red cell components. PMID- 11931384 TI - Fetuin in human bone marrow: detection in foetal tissue and patients with mastocytosis. AB - Fetuin, a foetal protein of unknown function, has been shown to be expressed in both the immune and nervous systems, especially during development. Here, we show for the first time, that fetuin is abundantly present in many cells of the foetal human bone marrow, but is restricted to cells of the monocytic lineage in the adult. Fetuin's immunoreactivity increased considerably in adult human bone marrow in some pathological conditions, particularly in mastocytosis and was also increased in bone marrows in some cases of acute leukaemias, especially in acute myeloid leukaemia. This increase in the presence of fetuin in neoplastic bone marrows is not reflected by an increased level of circulating fetuin. This last observation contradicts earlier suggestions that fetuin is specifically reduced in cancer patients. A consistent increase in fetuin immunoreactivity in bone marrow of most cases of mastocytosis, as demonstrated in this paper, could become a useful tool in the diagnosis of this disease. PMID- 11931383 TI - Expression of Set-alpha during morphogenesis of mouse lower first molar. AB - The detailed in situ expression pattern of the Set-alpha gene has been studied. Previously we showed that Set-alpha is a differentially expressed gene in the embryonic mouse mandible at day 10.5 (E10.5) gestational age. Cells expressing Set-alpha were widely distributed in both the epithelial and underlying ectomesenchymal cells at E10.5. At E12, they were slightly aggregated in an area where tooth germ of the lower first molar is estimated to be formed. At E13.5, Set-alpha was strongly expressed in the tooth germ. At the cap stage, Set-alpha was expressed in the enamel organ and dental papilla. At the bell stage, Set alpha was distinctly expressed in the inner enamel epithelial and dental papilla cells facing the inner enamel epithelial layer, which were intended to differentiate into ameloblasts and odontoblasts, respectively. Interestingly, Set alpha was also expressed in several embryonic craniofacial tissues derived from the ectoderm. This study is the first report that Set-alpha is distinctly expressed in the developing tooth germ, and suggests that Set-alpha plays an important role in both the initiation and the growth of the tooth germ, as well as in the differentiation of ameloblasts and odontoblasts. PMID- 11931385 TI - Localization of epidermal-type fatty acid binding protein in alveolar macrophages and some alveolar type II epithelial cells in mouse lung. AB - Almost all alveolar macrophages in the mouse lung were strongly immunoreactive for epidermal-type fatty acid binding protein. At the electron microscope level, the immunoreactive material was localized diffusely in the cytoplasm but not within the nucleus. A certain number of alveolar type II epithelial cells were also immunoreactive for the protein with variable immunointensity, while a substantial number of the type II cells were immunonegative. No immunoreactive interstitial fibroblasts were encountered. Based on the present findings, possible roles of epidermal-type fatty acid binding protein in the host-defence mechanism played by alveolar macrophages are suggested. PMID- 11931386 TI - DNA mismatch repair enzyme hMSH2 in malignant melanoma: increased immunoreactivity as compared to acquired melanocytic nevi and strong mRNA expression in melanoma cell lines. AB - Mutations in the mismatch DNA repair gene human MutS homologen 2 (hMSH2) are causative for microsatellite instability and carcinogenesis in various human tumours, including hereditary nonpolyposis colorectal cancer. Because microsatellite instability has been detected in malignant melanoma, we have investigated hMSH2 in melanocytic tumours. We found strong nuclear immunoreactivity for hMSH2 that was elevated in malignant melanoma and melanoma metastases as compared to acquired nevi. These findings suggest that increased genomic instability in malignant melanoma is associated with elevated protein levels of this DNA repair enzyme. hMSH2 is not exclusively regulated by proliferative activity in melanocytes, because there was no correlation between staining patterns of hMSH2 and the proliferation marker Ki-67. In contrast, immunoreactivity scores for hMSH2 and p53 were both upregulated in malignant melanocytic tumours. These findings support the concept that hMSH2 gene expression may be regulated in melanocytes by the p53 protein, as has been reported previously in other tissues. Using the reverse transcription-polymerase chain reaction, we detected strong hMSH2 mRNA expression in each of 8 melanoma cell lines analysed (highest amounts in SK-MEL-25 cells, lowest amounts in MML-I cells). In conclusion, our findings indicate that hMSH-2 may be of importance for genetic stability, tumorigenesis and progression of malignant melanoma. PMID- 11931388 TI - cDNA cloning and expression of the mouse Na/H antiporter (NHE-1) and a potential splice variant. AB - A cDNA for the Mus musculus Na/H exchanger-isoform 1 (NHE-1) was identified in a BALB/c myoblast library by its hybridization to rat NHE-1 sequences. Analysis of the clone showed it to display extensive homology with NHE-1 clones from other mammalian species; however, the region of interspecific homology was abruptly interrupted in the midst of the open reading frame by 166 bp of unrelated sequence. This extra sequence is likely to be an unspliced intron 9. Aside from the retained intron 9, the NHE-1 cDNA clone is otherwise fully processed, with all of the other ten introns removed and containing a poly(A) tract. From PCR results this variant represents a significant but minor population of NHE-1 RNAs. The variant message does associate with polysomes thereby suggesting it to be translated into protein. The location of the retained intron in the carboxy terminus of the protein is such that its translation would produce a protein predicted to be still capable of effecting Na and H translocation but whose regulatory features would be markedly altered. Amino acid sequence comparison of the mouse NHE-1 (derived from the fully processed message) with that of other mammalian species demonstrated two exceptionally divergent regions; the C terminal cytoplasmic tail (residues 750-790), containing a region of 6-8 contiguous acidic amino acids variably composed of aspartate and glutamate residues, and the N-terminal extracellular domain that includes an N-linked glycosylation site (residues 60-80). PMID- 11931387 TI - Structural changes of tRNA and 5S rRNA induced with magnesium and visualized with synchrotron mediated hydroxyl radical cleavage. AB - The structure of native yeast tRNA(Phe) and wheat germ ribosomal 5S RNA induced by different magnesium ion concentrations was studied in solution with a synchrotron mediated hydroxyl radical RNA cleavage reaction. We showed that very small amounts of Mg+2 can induce significant changes in the hydroxyl radical cleavage pattern of tRNA(Phe). It also turned out that a reactivity of tRNAz(Phe) towards *OH coincides with the strong metal binding sites. Because of the Mg ions are heavily hydrated one can suggest the strong correlation of the observed nucleosides reactivity in vicinity of Mg2+ binding sites with availability of water molecules as a source of hydroxyl radical. On the other hand the structure of wheat germ 5S rRNA is less sensitive to the hydroxyl radical reaction than tRNA(Phe) although some changes are visible at 4 mM Mg ions. It is probably due to the lack of strong Mg+2 binding sites in that molecule. The reactivity of nucleotides in loops C and D of 5S rRNA is not effected, what suggests their flexibility or involvement in higher order structure formation. There is different effect of magnesium on tRNA and 5S rRNA folding. We found that nucleotides forming strong binding sites for magnesium are very sensitive to X ray generated hydroxyl radical and can be mapped with *OH. The results show, that guanine nucleotides are preferentially hydrated. X-ray footprinting mediated hydroxyl radical RNA cleavage is a very powerful method and has been applied to studies of stable RNAs for the first time. PMID- 11931389 TI - Nucleotide sequence of BamHI family satellite DNA and its unit length polymorphism in bluegill sunfish Lepomis macrochirus. AB - We have isolated and sequenced a member of tandem repetitive DNA containing BamHI site (BamHI family satellite DNA) from bluegill sunfish Lepomis macrochirus. PCR amplification with specific primers was performed to define the size of unit length repeat of the BamHI family satellite DNA, revealing that there were two distinct size of DNA fragments (0.9 kb and 1.3 kb) in the PCR products. The longer fragment (1.3 kb) consisted of internal sub-duplication of shorter fragment (0.9 kb). We have compared the size of PCR products among four fish populations, and found that both fragments co-existed in one population whereas the longer fragment was dominant in other three populations. The results may reflect ongoing homogenization of satellite DNA type over a short evolutionary time scale. PMID- 11931390 TI - Functional analysis of a chromosomal arsenic resistance operon in Pseudomonas fluorescens strain MSP3. AB - We reported earlier about the detection of a chromosomally located arsenic operon (arsRBC) in a gram-negative bacterium Pseudomonas fluorescens strain MSP3, which showed resistance to elevated levels of sodium arsenate and sodium arsenite. The genes for arsenic resistance were cloned into the HindIII site of pBluescript vector producing three clones MSA1, MSA2 and MSI3 conferring resistance to sodium arsenate and arsenite salts. They were further sub-cloned to delineate the insert size and the sub-clones were designated as MSA11, MSA12 and MSI13. The sub-clone pMSA12 (2.6 kb) fragment was further packaged into EcoRI-PstI site of M13mp19 and sequenced. Nucleotide sequencing revealed the presence of three open reading frames homologous to the arsR, arsB and arsC genes of arsenic resistance. Three cistrons of the ars operon encoded polypeptides ArsR, ArsB and ArsC with molecular weights ranging approximately 12, 37and 24 kDa, respectively. These polypeptides were visualized on SDS-PAGE stained with Coomassie blue and measured in a densitometer. The arsenic resistance operon (arsRBC) of strain MSP3 plasmid pMSA12 consists of 3 genes namely, arsR--encoding a repressor regulatory protein, arsB--the determinant of the membrane efflux protein that confers resistance by pumping arsenic from the cells and arsC--a small cytoplasmic polypeptide required for arsenate resistance only, not for arsenite resistance. ArsB protein is believed to use the cell membrane potential to drive the efflux of intracellular arsenite ions. ArsC encodes for the enzyme arsenate reductase which reduces intracellular As(V) (arsenate) to more toxic As(III) (arsenite) and is subsequently extruded from the cell. The arsenate reductase activity was present in the soluble cytoplasmic fraction in E. coli clones. In the context of specified function of the arsenic operon encoded proteins, uptake and efflux mechanisms were studied in the wild strain and the arsenate/arsenite clones. The cell free filtrates of the arsenate clones (MSA11 and MSA12) obtained from P. fluorescens containing the arsC gene showed that arsenate reduction requires glutathione reductase, glutathione (GSH), glutaredoxin and ArsC protein. The protein was purified in an active form and a spectrophotometric assay was developed in which the oxidation of NADPH was coupled to reduction of arsenate. The molecular weights and the location of the polypeptides were obtained from Coomassie stained SDS-PAGE of extracellular and intracellular fractions of the cells. PMID- 11931391 TI - Genome wide oscillations in expression. Wavelet analysis of time series data from yeast expression arrays uncovers the dynamic architecture of phenotype. AB - A reanalysis of expression arrays in yeast cells synchronized by alpha factor blockade or through the use of temperature sensitive mutants uncovered a genome wide pattern of oscillations in mRNA concentrations. Using wavelet decomposition as a signal processing technique and enhancement strategies borrowed from image processing, noise and trends in the Stanford yeast cell cycle data were partitioned away from time series profiles to uncover genome-wide oscillations in expression. These oscillations which were typically of cell cycle or half cell cycle duration, 40 and 80 minutes in the Stanford data set suggest that there are large-scale temporal structures and high frequency oscillations in mRNA levels through the cell cycle. Wavelet decomposition, which acts like a band pass filter bank, was used to determine where most of the power appeared in the decomposition. The approximately 40-min oscillation is mirrored in continuous chemostat cultures. In these cultures, metabolic synchrony involving an unknown proportion of the transcriptome can be monitored by measurement of oxygen consumption and can be sustained for weeks. These 40-min oscillations are stable and precise with coefficients of variation less than 1% for both period and amplitude. The hypothesis that high and low amplitude oscillations are a ubiquitous property of the genetic regulatory circuitry was supported by the observation of period doubling bifurcations in the distribution of population doubling times in yeast. PMID- 11931392 TI - Computational cell biology in the post-genomic era. AB - Rapid accumulation of biological data from novel high throughput technologies characteristic of genomic and proteomic research as well as advances in more traditional biological disciplines are leading to wider use of detailed and complex computational models of cell behavior. These models address a variety of dynamic intracellular processes ranging from interactions within a gene regulation network to intracellular and intercellular signal transduction. This review focuses on the current trends in computation cell biology, particularly emphasizing the role of experimental validation. The recent successes and future challenges facing computational cell biology are also discussed. PMID- 11931393 TI - Identification of human, mouse and rat PPP1R14A, protein phosphatase-1 inhibitor subunit 14A, & mapping human PPP1R14A to chromosome 19q13.13-q13.2. AB - Three novel cDNAs encoding putative protein phosphatase-1 inhibitory protein (PPP1R14A) were isolated from human, mouse and rat. The human, mouse and rat PPP1R14A proteins were all of 147 amino acids sharing about 90% sequence identity to porcine CPI17, an inhibitor of protein phosphatase-1 holoenzyme. The protein sequence 33RHARVTVK40, which is important for the inhibitory potency of porcine CPI17, was conserved in mammalian PPP1R14A. Northern blot analysis showed that human PPP1R14A was highly expressed as a 600 bp transcript in heart, prostate, testis, ovary, colon, small intestine and pancreas, lowly in brain, placenta, skeletal muscle and spleen and lung. The distribution of mouse and rat PPP1R14A was more specific and different from that of human PPP1R14A, abundant in lung, moderate/abundant in testis, moderate in brain and low in heart. In addition, we mapped human PPP1R14A to chromosome 19q13.13-q13.2 by radiation hybrid mapping, and determined that the human PPP1R14A gene spanned a 5.1-kb region and consisted of four exons and three introns. PMID- 11931394 TI - Another step on the path to better TB therapies. PMID- 11931395 TI - Sex differences in tuberculosis in Hong Kong. AB - SETTING: The Hong Kong Tuberculosis and Chest Service, Department of Health, Hong Kong. OBJECTIVE: To examine sex differences in the rate and clinical manifestations of tuberculosis in Hong Kong. DESIGN: Notification rates of tuberculosis during the past five decades were obtained from the Hong Kong Tuberculosis and Chest Service, Department of Health, Hong Kong. In addition, all patients registered with the Chest Service for treatment of tuberculosis in 1996 were studied. RESULTS: The rate of tuberculosis during the past five decades was consistently higher in men than in women, irrespective of age group. The sex difference in rates was highest among those aged over 60 years. In 1996, a higher proportion of women had extra-pulmonary tuberculosis than men; the main site of involvement was the lymph nodes. More women completed treatment at 12 months and fewer women missed treatment appointments. A higher proportion of men had relapse pulmonary disease that was more extensive, a history of previous default from treatment and co-morbid illnesses. CONCLUSION: There are sex differences in the rates and clinical manifestations of tuberculosis in Hong Kong. Study of sex differences is essential for targeting prevention programmes at groups at higher risk. PMID- 11931396 TI - Contact examination for tuberculosis in Hong Kong is useful. AB - SETTING: The tuberculosis programme of the Hong Kong Government Tuberculosis and Chest Service. OBJECTIVE: To determine the outcome of examination of household contacts in Hong Kong. DESIGN: A retrospective cohort study of all household contacts of 970 randomly selected index cases from a total of 5757 registered for treatment with the Chest Service. RESULTS: Of 2678 household contacts (three/index case) identified, 90% were examined; 41 active cases were found, at a rate of 1720/100000 (95%CI 1238-2329). The rate was highest among two extremes of age, 3604/100000 (95%CI 990-3615) in children < or = 5 years and 3347/100000 (95%CI 1456-6489) in those >60 years of age. Contacts of index cases whose sputum smear and culture were positive had the highest rate of disease, 2904/100000 (95%CI 1669-4673); but contacts with negative bacteriology also had a high rate of 1478/100000 (95%CI 678-2789). Active cases identified through contact tracing could be source cases rather than secondary cases. Eight per cent of children aged < or = 5 years had positive tuberculin reactions; as BCG vaccination is given to all newborns, with 99% coverage in Hong Kong in the past 30 years, it was difficult to estimate the rate of infection in these children. CONCLUSION: In Hong Kong, an area with an intermediate burden of tuberculosis, contact investigation is a very useful procedure for active case finding. PMID- 11931397 TI - Treatment adherence of 717 patients with tuberculosis in a social security system hospital in Istanbul, Turkey. AB - SETTING: Tuberculosis clinic in a teaching hospital run by the social security system, Istanbul, Turkey. OBJECTIVE: To determine risk factors associated with treatment adherence. DESIGN: Seven hundred and seventeen patients who commenced treatment for tuberculosis in our clinic from May 1991 to May 1997 were evaluated retrospectively with respect to treatment adherence. Factors with an effect on treatment adherence were investigated. RESULTS: Sputum conversion was achieved in 88.9% of the cases within the first 2 months of treatment. Seven patients died during treatment; of the 710 patients remaining, 106 (14.9%) were non-adherent. This rate decreased from 34.4% in 1991 to 2.0% in 1997. In multi-variate logistic regression analysis, only previous treatment history for tuberculosis was related to non-adherence; treatment adherence rate in new cases was 88.9%, while it was 66.7% in previously treated cases (P < 0.001). CONCLUSION: In new cases, a treatment adherence rate of 88.9% can be considered satisfactory. However, in previously treated cases, an adherence rate of 66.4% must be considered unsatisfactory. Previously treated cases in particular should therefore receive directly observed treatment. PMID- 11931399 TI - Prevalence of resistance to anti-tuberculosis drugs: results of the 1998/99 national survey in Italy. AB - OBJECTIVE: To determine the prevalence of resistance to the main anti tuberculosis drugs in newly and previously treated tuberculosis patients in Italy and to evaluate the contribution of foreign-born and human immunodeficiency virus (HIV) positive cases to drug resistance. METHODS: Methods and definitions were derived from the WHO/IUATLD Global Project on Anti-tuberculosis Drug Resistance Surveillance. Univariate and multivariate analysis was used to study prevalence rates of drug resistance in risk groups. RESULTS: In a national survey in Italy, 810 initial isolates of Mycobacterium tuberculosis (683 from new cases, 115 from retreatment cases and 12 from patients whose treatment history was unknown/dubious) were analysed. Low prevalence of drug and multidrug resistance was found in the new cases (isoniazid 2.9%; rifampicin 0.8%; multidrug resistance 1.2%; any drug resistance 12.3%). The prevalence of resistance to isoniazid and rifampicin was significantly higher in immigrants and HIV-positive subjects, respectively. A high prevalence of drug resistance was found in cases with previous treatment failure or default (isoniazid 5.2%; rifampicin 4.3%; multidrug resistance 36.5%; any drug resistance 61.7%). RECOMMENDATIONS: Special efforts are necessary to monitor trends in drug resistance and to ensure favourable treatment outcomes among immigrants and HIV-positive tuberculosis cases. PMID- 11931398 TI - Rifapentine and isoniazid in the continuation phase of a 6-month regimen. Final report at 5 years: prognostic value of various measures. AB - SETTING: Clinical trial in 672 patients with newly diagnosed pulmonary tuberculosis in Hong Kong. After an initial 2 months of a four-drug intensive phase consisting of thrice-weekly streptomycin, isoniazid, rifampicin and pyrazinamide (SHRZ), a random allocation was made to a continuation phase of once weekly 600 mg rifapentine + 15 mg/kg isoniazid (HRp1), HRp1 given in 2 of every 3 weeks (HRp1.2/3), or to thrice-weekly isoniazid + rifampicin (HR3), the standard treatment in Hong Kong. OBJECTIVE: Final report evaluating adverse events (46 relapses and one failure) after 5 years and the prognostic influence of various factors. METHODS: Kaplan-Meier analysis of adverse events and Cox proportional hazards analysis of prognostic factors. RESULTS: The two rifapentine regimens, HRp1 and HRp1.2/3 had similar final rates of adverse events of 10.8% and 11.7%, respectively, compared to 4.2% for the HR3 regimen (P = 0.02 and 0.009, respectively). In the initial univariate proportional hazards analysis, adverse events were significantly related to the regimen, age, sex, pretreatment radiographic extent of disease and cavitation, and also to sputum culture at 2 months. In the final multivariate analysis, after step-wise removal of non significant factors, adverse events were related only to the regimen, patients' sex and pretreatment radiographic extent of disease. Elderly male patients were more at risk of an adverse event, as were those with more severe disease. Adverse events occurred at life table rates of 9.0% in patients with drug-sensitive strains and in 8.9% of those with initially isoniazid-resistant organisms at 5 years. CONCLUSIONS: The two rifapentine regimens were unsatisfactory because of their high incidence of adverse events. Isoniazid appeared not to contribute to preventing relapse. Further studies with increased rifapentine dosage are necessary. PMID- 11931400 TI - Quality control of sputum smear examination in Cebu Province. AB - SETTING: Rural Health Units (RHUs) in Cebu Province, Philippines. OBJECTIVE: To assess and improve the on-going quality control of tuberculosis activities in Cebu Province. METHODS: During the period of the study, from January 1996 to December 1997, positive slides (100% in 1996 and 20% or 100% in 1997) and 20% of negative slides selected from all of the slides examined at the RHUs were assessed on the quality of smear preparation. The readings were blindly cross checked by the provincial assessors. RESULTS: In 1997, 1) 90% of RHUs participated in the quality control activity; 2) the proportion of good quality smears increased markedly; and 3) the false positive and false negative rates did not change during the period, but most of the false positives were observed among the scanty positives of the field reading, and no false negatives were noted among the heavily positive slides. CONCLUSION: Although the false positive and negative rates did not decrease during the period, the quality control procedure resulted in marked improvements in smear preparation, a high participation of RHUs in the quality control programme and the elimination of large discrepancies between readers on positive slides in 1997. It is considered to have significantly improved the NTP's smear microscopy service in Cebu Province. PMID- 11931401 TI - Quality assessment of sputum transportation, smear preparation and AFB microscopy in a rural district in Malawi. AB - SETTING: Ntcheu District, Central Region of Malawi. OBJECTIVES: To assess 1) the feasibility of introducing simple internal quality control procedures for acid fast bacilli (AFB) microscopy, and 2) the quality of the district sputum smear microscopy service. DESIGN: A simple internal quality control system was piloted in which district laboratory staff assessed: 1) specimen suitability, 2) time between sputum submission and smear examination, 3) smear preparation and staining, and 4) microscopy. Actual times for processing specimens were compared with recommended times. External quality validation was carried out. RESULTS: Of 4805 sputum specimens: 1) documentation was complete in 95%, 2) 93% reached the laboratory within 7 days of collection, 3) 96% of smears were well prepared and stained, and 4) 97% concordance (96.4% smear-positive and 97.6% smear-negative) was demonstrated when 208 smears were re-examined by a second technician. The aggregate index of reliability was 86%. The mean time spent on microscopic examination was 3.8 minutes, compared with the recommended time of 10 minutes. When all smears from 164 patients were assessed externally, 98.2% concordance (98.1% smear-positive and 98.2% smear-negative) was demonstrated. False smear negative and smear-positive rates were less than 2% each. CONCLUSION: District laboratory staff were able to incorporate simple quality control procedures for AFB microscopy into their routine practice, resulting in a reliable service. The lessons learnt are widely relevant and potentially useful for implementation of a national quality assurance scheme. PMID- 11931402 TI - Pathology and causes of death in a group of 128 predominantly HIV-positive patients in Botswana, 1997-1998. AB - BACKGROUND: Little is known about causes of death in countries of southern Africa seriously affected by the HIV/AIDS epidemic. METHODS: After obtaining informed consent, autopsies were performed on 128 mainly hospitalised adults in Francistown, Botswana, between July 1997 and June 1998. Criteria for case selection included those who died before a diagnosis could be established, those whose condition deteriorated unexpectedly during hospitalization, and those who had respiratory disease. This represented 14% of adult medical patients who died in hospital during the study period. RESULTS: Of the 128 patients, 104 (81%) were HIV-positive. Among HIV-positive patients, the most common pathologic findings were tuberculosis (TB) (40%), bacterial pneumonia (23%), Pneumocystis carinii pneumonia (11%), and Kaposi's sarcoma (11%); these conditions were the cause of death in 38%, 14%, 11%, and 6%, respectively. Of the 40 pulmonary TB cases, 90% also had disseminated extra-pulmonary TB. Chest radiology could not reliably distinguish the pathologies pre-mortem. CONCLUSIONS: TB was the leading cause of death in our series of HIV-positive adults in Botswana, selected towards those with chest disease; in most, it was widely disseminated. Bacterial pneumonia also played an important role in mortality. Pneumocystis carinii pneumonia was present, but relatively uncommon. PMID- 11931403 TI - Predictors of outcome in patients with tuberculous meningitis. AB - OBJECTIVE: To assess predictors of mortality and neurological sequelae in patients with tuberculous meningitis (TBM). METHODS: Patients with TBM treated at 12 university hospitals in Turkey between 1985 and 1997 were evaluated using a standardised protocol applied retrospectively. Variables associated with hospital mortality as well as with the presence of neurological sequelae at 6 months were determined using logistic regression models. RESULTS: Four hundred and thirty four patients between the ages of 13 and 83 years (mean 33 years) were evaluated. Sixty-eight per cent of these patients presented with Medical Research Council Stage II or III. One hundred and one patients (23.3%) died and 67 (27%) of evaluable survivors had neurological sequelae. In multi-variable analysis, convulsion (OR 3.3, 95%CI 1.2-9.0, P = 0.02), comatose mental status (OR 6.0, 95%CI 3.6-10.2, P = 0.01), and delayed or interrupted treatment (OR 5.1, 95%CI 2.4-11.2, P = 0.01) were shown to be predictors for mortality. The presence of extra-meningeal tuberculosis (OR 2.1, 95%CI 1.1-4.2, P = 0.035), cranial nerve palsy (OR 2.6, 95%CI 1.4-4.2, P = 0.01), hemiparesia/focal weakness (OR 9.3, 95%CI 3.8-22.6, P = 0.01), hemiplegia/multiple neurological deficit (OR 7.1, 95%CI 2.14-23.38, P = 0.01) and drowsiness (OR 4.2, 95%CI 2.04-8.82, P = 0.01) were independent predictors of neurological sequelae at 6 months following hospital discharge. CONCLUSION: The results of this study emphasise the importance of prompt and uninterrupted anti-tuberculosis therapy for tuberculous meningitis. The presence of seizures or coma on admission to hospital are important predictors for mortality, while the presence of focal neurological signs is a predictor for persistent neurological sequelae in survivors. PMID- 11931404 TI - Paradoxical responses in a cohort of HIV-1-infected patients with mycobacterial disease. AB - SETTING: Paradoxical worsening or relapse of opportunistic infections has been described after initiation of highly active anti-retroviral therapy (HAART) in human immunodeficiency virus (HIV) infected patients. DESIGN: Retrospective study of a group of 33 HIV-infected patients with mycobacterial disease analysing the incidence and characteristics of patients with and without paradoxical response after starting HAART and/or mycobacterial treatment. RESULTS: Nine patients in the group had paradoxical response. No significant difference of baseline characteristics was observed in these patients. The decrease in viral load was significantly greater among patients with paradoxical response than in patients without. CONCLUSION: No clinical difference was found in the evolution of HIV infected patients with mycobacterial disease after the resolution of the episode of paradoxical response. PMID- 11931405 TI - Underdiagnosis of chronic obstructive pulmonary disease in Northern Sweden. AB - OBJECTIVE: To assess underdiagnosis of chronic obstructive pulmonary disease. MATERIAL AND METHODS: Two cross-sectional studies of respiratory symptoms and diseases in two population samples of the same age living in the same areas in northern Sweden were performed 6 years apart. In 1986, 5698 (86%) out of 6610 subjects aged 35-36, 50-51 and 65-66 years responded to a postal questionnaire. In 1992 an identical study was performed, and 5617 subjects (87%) out of 6434 responded. Lung function measurements were performed in stratified samples. RESULTS: Of the subjects diagnosed with chronic bronchitis only 25% in 1986 and 23% in 1992 had been diagnosed prior to the study as having chronic bronchitis, emphysema or chronic obstructive pulmonary disease (COPD). Chronic airflow limitation (CAL), used as a surrogate variable for COPD and defined as FEV1/VC <70% and FEV1 <80% of predicted value, was found in 171 subjects in 1986-1987 (12% of the examined subjects), and 166 subjects in 1993-1994 (11%). In 1986 1987, 26% of the subjects with CAL had been diagnosed as having chronic bronchitis or emphysema prior to the survey, while a diagnosis of either asthma, chronic bronchitis or emphysema, or use of asthma medicines, was found in 58%. The corresponding figures in 1993-1994 were 31% and 63%, respectively. The great majority of the subjects with CAL had recurrent wheeze, dyspnoea and chronic productive cough. CONCLUSION: Approximately 60% of the subjects with chronic airflow limitation had been diagnosed prior to the survey as having asthma, chronic bronchitis or emphysema, or were using asthma medicines. The results were similar in 1986-1987 and 1993-1994. PMID- 11931406 TI - Increase in tuberculosis incidence among the staff working at the Tuberculosis Demonstration and Training Centre in Addis Ababa, Ethiopia: a retrospective cohort study (1989-1998). AB - The incidence rate of tuberculosis was studied among the staff of the Tuberculosis Demonstration and Training Centre (TDTC) of Addis Ababa between 1989 and 1998, by reviewing all clinical charts of the 175 staff members for evidence of tuberculosis. During the study period, 24 cases of tuberculosis were diagnosed, including 12 who were bacteriologically confirmed. The incidence rate of tuberculosis increased from 1695 per 100000 person-years (py) in 1989 to 5556/100000 py in 1998 (test for trend, P < 0.001). Urgent measures are required for better protection of the staff from human immunodeficiency virus infection and tuberculosis. PMID- 11931407 TI - Analysis of data from individuals demonstrates high discordance between IFN-gamma assays and PPD-tuberculin skin tests used in screening for tuberculosis infection. PMID- 11931408 TI - What motivates lay volunteers in high burden but resource-limited tuberculosis control programmes? Perceptions from the Northern Cape province, South Africa. AB - SETTING: The Northern Cape province, Republic of South Africa. OBJECTIVES: To explore factors that motivate lay volunteers to join tuberculosis (TB) control programmes in high burden but resource-limited settings. DESIGN: A qualitative study consisting of three focus group discussions and a documentary review of the records of 347 lay volunteers involved in the tuberculosis programme in the Northern Cape province of South Africa. Additional data were also collected in a cross-sectional study that involved questionnaire interviews with 135 lay volunteers. SUBJECTS: Lay volunteers in the TB programme. One focus group discussion was also carried out with youth not involved in the TB programme. RESULTS: Volunteers do not receive any monetary incentives in the TB programme in the Northern Cape province, but due to the high level of unemployment in this setting, hope for eventual remuneration was found to be the strongest factor motivating youth to join the programme. The study found attrition rates among volunteers to be high (22% had dropped out of the programme within one year of joining); 75% of the dropouts gave loss of interest and a desire for paid work as the reasons for leaving the TB programme. Other motivating factors identified included altruism, a need to find something to do with one's spare time, gaining work experience, and the novelty of the community-based TB programme. CONCLUSION: In the absence of monetary incentives, attrition rates of lay volunteers may be high and this can threaten the effectiveness of community-based TB programmes. In resource-limited settings, it is important to identify and implement appropriate alternative incentives that could motivate lay persons in order to sustain community participation in high TB burden areas. PMID- 11931409 TI - Use of a computerized tuberculosis register for automated generation of case finding, sputum conversion, and treatment outcome reports. AB - SETTING: Tuberculosis (TB) rates in southern Africa have increased dramatically in recent years. Provision of accurate data for surveillance, program management, and supervision is increasingly essential. OBJECTIVE: To develop software that would provide more efficient collection, compilation, and analysis of TB data on an ongoing basis. DESIGN: The 'Electronic TB Register' is a user-friendly, Epi Info based software program based on the WHO/IUATLD format of recording and reporting. Individual records from the TB registry are entered in a program that provides interactive support. The software provides several patient management and supervision functions, such as lists of defaulters. Finally, it generates standard quarterly and annual reports on case-finding, sputum conversion, and cohort analysis, and provides graphs of trends and maps of TB indicators. RESULTS: The 'Electronic TB Register' software has been successfully implemented in five pilot projects in southern Africa. User acceptance has been high and quality of data has improved, although timeliness remains unchanged. Factors critical for success include a functioning, paper-based system, involvement of staff from the TB program, health information systems, and health facilities, ongoing training, and backup support. CONCLUSIONS: The 'Electronic TB Register' is a potentially powerful tool for surveillance, management, and supervision for countries with well-functioning paper-based recording and reporting systems. PMID- 11931410 TI - Effect of screening of immigrants on tuberculosis transmission. AB - SETTING: In The Netherlands all immigrants from highly endemic countries undergo obligatory entry screening by X-ray, followed by voluntary half-yearly screening for 2 years. OBJECTIVE: To estimate the contribution of screening of immigrants to reductions in tuberculosis transmission. DESIGN: All bacteriologically confirmed tuberculosis patients diagnosed between 1994 and 1999 with mycobacterial isolates exhibiting identical DNA fingerprints were assigned to clusters. Clusters were assumed to be a consequence of recent transmission and to have had the first patient as their source case. Among patients with pulmonary tuberculosis from highly endemic countries, the characteristics of source patients were compared with those of non-clustered patients. RESULTS: Of the 1438 selected patients, 187 (13%) were the first in a cluster, 386 (27%) were in a cluster but were not the first case, and 865 (60%) were not clustered. Independent risk factors for being the first in a cluster were young age, nationality, early year of diagnosis, longer duration of treatment and no concurrent extra-pulmonary tuberculosis. In univariate analysis, passively detected patients and patients with a long duration of stay in The Netherlands were more likely to be the first in a cluster than patients detected by screening and recent arrivals, respectively. However, these variables were strongly associated. CONCLUSION: Screening of immigrants can reduce tuberculosis transmission. This effect is in part due to confounding by duration of stay. PMID- 11931411 TI - The impact of immigration on the elimination of tuberculosis in The Netherlands: a model based approach. AB - OBJECTIVE: To determine whether elimination of tuberculosis in the Dutch population can be achieved by the year 2030, taking into account the impact of immigration. METHODS: The incidence of tuberculosis (all forms) in the period 1970 to 2030 was estimated using a life-table model for the Dutch population without the impact of immigration. The influence of immigration on tuberculosis incidence among the Dutch was modelled using four immigrant scenarios, distinguished by the assumed contact rate between immigrants and the Dutch population, and by different projections (middle, upper) of the future size of the immigrant population in The Netherlands. RESULTS: The incidence of smear positive tuberculosis among the Dutch is projected to be 1.4 per million in the scenario without the influence of immigrant cases, and ranging from 3.8 to 11.8 per million in the four immigrant scenarios. In all immigrant scenarios, the prevalence of tuberculosis infection will continue to decline and be less than 1% by the year 2030. At least 60% of Dutch tuberculosis cases in the year 2030 are expected to be the result of transmission from a foreign source case. CONCLUSION: Using a prevalence of tuberculosis infection of less than 1% as the elimination criterion, tuberculosis will probably be eliminated from the indigenous Dutch population by 2030. However, the incidence of smear-positive tuberculosis is expected to remain higher than 1 per million, and the majority of new tuberculosis cases among the Dutch may be attributable to recent infection from a foreign source case. PMID- 11931412 TI - Chest radiograph abnormalities associated with tuberculosis: reproducibility and yield of active cases. AB - SETTING: Tertiary care referral centre specialising in respiratory diseases. OBJECTIVES: Chest radiography is a major screening and diagnostic tool for tuberculosis (TB). We evaluated the reproducibility of a radiographic classification system for screening for active TB of immigration applicants to Canada. We also evaluated the validity of this classification system for detection of prevalent active TB among the screened applicants, as well as tuberculin-positive close contacts and symptomatic patients. METHODS: Reproducibility was assessed by re-reading a randomly selected 10% sample of screening chest films. Validity was estimated from the final clinical and microbiologic diagnosis of patients undergoing detailed clinical evaluation. RESULTS: Inter-reader agreement using five broad categories was moderate (kappas of 0.44-0.56), while intra-reader agreement was substantial (kappas of 0.59 0.72). After adjustment for age and patient group, the adjusted odds of active tuberculosis, relative to normal or minor findings or granulomas, for fibronodular changes was 10.2 (95% confidence interval [CI] 3.2-33), for mass or pleural effusion it was 11.6 (95%CI 3.6-37), and for parenchymal infiltrate it was 46.1 (95%CI 18-117). Among tuberculin-positive close contacts, the probability of active tuberculosis was more than 50% if the radiographs showed any mass, pleural disease, or parenchymal infiltrates. CONCLUSION: A simple classification of TB-related chest radiographic abnormalities into five broad categories had moderate to substantial reproducibility of readings, with reasonable validity. PMID- 11931413 TI - A retrospective study for the outcome of pulmonary resection in 49 patients with multidrug-resistant tuberculosis. AB - BACKGROUND: Outcome of treatment using only chemotherapy for multidrug-resistant tuberculosis is usually considered not to be satisfactory. The combination of chemotherapy with surgical treatment has been producing higher acid-fast bacilli (AFB) negative sputum conversion and longer survival rates. This treatment strategy may be the solution for patients with multidrug-resistant tuberculosis. MATERIALS AND METHODS: A retrospective review was performed of the medical records and laboratory findings of 49 patients with multidrug-resistant tuberculosis among 130 patients who underwent pulmonary resection for pulmonary tuberculosis between January 1995 and December 1999 at National Masan Tuberculosis Hospital, Korea. RESULTS: The mean number of drugs to which the patients were resistant was 4.5. Patients had a mean age of 35 years. Cavitary lesions on plain chest X-ray were shown in 43/49 patients (87.8%); 31 had positive sputum cultures preoperatively (63.3%). The surgical techniques used were as follows: 12 pneumonectomies, 28 lobectomies, seven lobectomies with segmentectomies or wedge resections, one wedge resection and one cavernoplasty. The AFB negative sputum conversion rate was 93.5% with continuous postoperative chemotherapy. There were no deaths after surgery. Postoperative complications that developed were six cases of air leakage over a week, one of postoperative bleeding and one of wound infection. CONCLUSIONS: This study demonstrated the effectiveness of pulmonary resection with postoperative chemotherapy in cases of multidrug-resistant tuberculosis. Although there are different ideas about indications for surgery and the length and content of postoperative drug regimens, pulmonary resection should be considered an effective measure in combination with chemotherapy for treatment of multidrug-resistant pulmonary tuberculosis. PMID- 11931414 TI - Mycobacterium tuberculosis typing: usefulness of DRE-PCR to confirm cross contamination in the mycobacteriology laboratory of a general reference hospital for AIDS. AB - In this study two molecular typing methods, a simple double repetitive element PCR-based assay and the standardized restriction fragment length polymorphism (RFLP), were used to confirm cross-contamination in the mycobacteriology laboratory. Clinical specimens from 12 patients, submitted for acid-fast bacilli stain smear and processed for culture in Lowenstein-Jensen on the same day, resulted in positive bacterioscopy (+++) and confluent growth only for one of the patients. The specimens from all the other patients but two were smear-negative and culture-positive, with one or two colonies. None of them had clinical symptoms and radiological findings for active tuberculosis (TB). The suspicion of false-positive cultures arose when a health care worker who had had a PPD skin test conversion, claimed to be healthy and had no TB symptoms, was found to have a positive sputum culture. DRE-PCR demonstrated that all nine cultures typed belonged to one cluster, further confirmed by RFLP. Although DRE-PCR has been found to be poorly reproducible, it has enough discriminatory power to be useful for rapid epidemiological investigation in selected settings. PMID- 11931415 TI - sICAM-1 as a serum marker in the diagnosis and follow-up of treatment of pulmonary tuberculosis. AB - OBJECTIVE: To detect the value of sICAM-1 in the diagnosis and follow-up of treatment of tuberculosis. DESIGN: sICAM-1 levels were evaluated before and after treatment in 30 patients with pulmonary tuberculosis, only before treatment in five patients with pneumonia, five with lung cancer, and five with bronchiectasis, and in 10 healthy volunteers. RESULTS: sICAM-1 levels were as follows: 436.2 +/- 194.4 ng/ml in patients with pulmonary tuberculosis, 274 +/- 32.1 ng/ml in lung cancer patients, 268 +/- 41.9 ng/ml in bronchiectasis patients, 199.6 +/- 43.1 ng/ml in pneumonia patients, and 146.5 +/- 20.2 ng/ml in healthy individuals. sICAM-1 levels of tuberculosis cases before treatment were higher than in both the healthy group and in all the other groups. The levels in the healthy group were lower than in all other groups. CONCLUSION: The cut-off point (298 ng/ml) obtained by adding the standard deviation to the mean sICAM-1 value of patients without tuberculosis had 83.3% sensitivity, 86.6% specificity and 84.4% accuracy in differentiating pulmonary tuberculosis from other pulmonary diseases. sICAM-1 can be used as an auxiliary marker in the diagnosis of pulmonary tuberculosis. PMID- 11931416 TI - Rifampin and isoniazid resistance associated mutations in Mycobacterium tuberculosis clinical isolates in Seville, Spain. AB - The susceptibility phenotypes of 964 clinical isolates of Mycobacterium tuberculosis were studied over a 7-year period in Seville, Spain. Thirty-eight (3.9%) strains were rifampin (RMP) resistant, 79 (8.2%) were isoniazid (INH) resistant and 22 (2.3%) were resistant to at least both antimicrobials (multidrug resistant, MDR). We studied the mechanisms of resistance to these drugs in 94 resistant clinical isolates of M. tuberculosis using three molecular methods: 1) PCR-single strand conformation polymorphism (SSCP) analysis, 2) RFLP analysis using B1/B2 primers, and 3) sequence analysis. Five different mutations were detected in the rpoB gene: Ser531-->Leu (72.3%), His526-->Asp (12.8%), Asn518- >Ser (2.1%), Gln513-->Leu (2.1%) and a nine-nucleotide deletion (2.1%). In the case of resistance to INH, four different mutations in the katG gene were detected, Ser315-->Thr (58.0%), Ser315-->Leu (2.9%), partial deletion (5.8%) and Ile304-->Val (1.4%), while in the inhA regulatory region the only mutation was the nucleotide substitution C209T (4.3%). No mutation was found in the ahpC promoter. PMID- 11931417 TI - Investigation of cross-resistance between rifampin and rifabutin in Mycobacterium tuberculosis complex strains. AB - In order to indicate the cross-resistance between rifampin (RMP) and rifabutin (RBU), minimal inhibitory concentrations (MICs) of RBU were investigated in 50 Mycobacterium tuberculosis strains. The MIC values of 25 RMP-susceptible (to 2 microg/ml) and 25 RMP-resistant (to 2 microg/ml) M. tuberculosis strains against RBU were determined by the Bactec TB 460 system. All of the RMP-susceptible strains were also susceptible to RBU (MIC < or = 1 microg/ml). Three out of 25 (12%) RMP-resistant strains were determined as susceptible to RBU. The high level cross-resistance (88%) obtained in this study highlights the importance of testing susceptibility to RBU prior to its inclusion in the tuberculosis treatment regimens at the Istanbul Faculty of Medicine. PMID- 11931418 TI - Cross-resistance between rifampicin and KRM-1648 is associated with specific rpoB alleles in Mycobacterium tuberculosis. AB - KRM-1648 resistant Mycobacterium tuberculosis strains were identified from a collection of rifampicin-resistant strains. Several strains had novel rpoB gene mutations in codons 512, 529 and 533 of the rpoB gene. The strains with mutations in codons 526 or 531, major mutation sites in rifampicin-resistant M. tuberculosis, were resistant to KRM-1648. Also, the strains with other mutations in the rpoB gene that were initially susceptible to KRM-1648 were prone to developing KRM-1648 resistance after further mutation. Thus, KRM-1648 is unlikely to be useful for the treatment of rifampicin-resistant tuberculosis. PMID- 11931419 TI - Private pharmacies in tuberculosis control--a neglected link. AB - In most settings in India, private pharmacies dispense prescriptions for anti tuberculosis drugs made out by private practitioners. In a cross-sectional study, we assessed the dispensing practices for tuberculosis and knowledge about the national tuberculosis programme of 300 pharmacies. In all, 2800 prescriptions were dispensed monthly by the pharmacies. Doctors' prescriptions were for durations of several months, but half of the patients bought drugs one dose at a time for self-administration. This practice might promote drug resistance. Although 95% of pharmacists were not aware of the existence of the tuberculosis programme, the majority (97%) were willing to learn and contribute towards tuberculosis control. The need and the potential of private pharmacies for participation in tuberculosis control are highlighted. PMID- 11931420 TI - Utility of fluoroquinolones in multidrug-resistant tuberculosis (MDR-TB)--a balanced view? PMID- 11931421 TI - Periodic or routine susceptibility testing in tuberculosis control. PMID- 11931422 TI - Relation of grading of sputum smears with clinical features of tuberculosis patients in routine practice in Sudan. AB - SETTING: Health services in Khartoum, Red Sea, Gadarif, Kassala, Bahr el Jebel, Gezira and North Kordofan States of Sudan. OBJECTIVE: To investigate the association of presenting symptoms and clinical history with grade of sputum smear positivity in tuberculosis patients. METHODS: The 5338 tuberculosis suspects aged 15 to 49 years who consulted between March 1998 and March 1999 underwent sputum smear microscopy, with smears graded by a standardised method. Patients who gave consent had a structured interview concerning the nature and duration of their symptoms and whether they had a sick relative at home. Statistical analysis determined the association of symptoms and home characteristics with the results of smear examination. RESULTS: A total of 514 suspects (9.6%) demonstrated acid-fast bacilli: 45 (8.8%) grade +1, 167 (32.5%) grade +2 and 302 (58.8%) grade +3. Most of the patients had a duration of symptoms of 6-9 weeks, while 100 had symptoms of more than one year's duration, many (n = 47) with other chronic respiratory diseases. Multiple chest symptoms were positively, and duration of symptoms negatively associated with grade of positivity (P = 0.018 and 0.028, respectively). A high grade of smear positivity (+3) and the longest duration of symptoms, among those reporting symptoms of less than one year's duration, were significant predictors of the presence of a sick household member (OR = 1.99, 95%CI 1.12-3.55; OR = 1.004, 95%CI 1.000-1.008). CONCLUSION: This study demonstrates a relation between the grade of smear positivity and the clinical features of tuberculosis patients. PMID- 11931423 TI - Is the distance a patient lives from hospital a risk factor for death from tuberculosis in rural South Africa? AB - SETTING: Tuberculosis programmes that rely on district hospitals for diagnosis and initiation of treatment may disadvantage those living furthest away. We present an analysis of such a programme in rural South Africa to see whether those living furthest from the hospital were at greatest risk of dying from tuberculosis. METHODS: All patients diagnosed and treated for tuberculosis in three health districts in 1997 and 1998 were included. An estimate of the distance each patient travelled to get to the hospital was obtained. The distances the patients travelled were categorised into four groups. The furthest distance patients could reasonably be expected to travel to get to their nearest hospital was estimated as 60 km. Outcomes of treatment were recorded using standard definitions. The mortality of patients in each of the four groups was compared. RESULTS: Of 1187 patients started on treatment for tuberculosis in the hospitals, 877 (74%) were known to be alive at the end of treatment, whereas 158 (13%) had died. Distance travelled was a risk factor for death, but only amongst those travelling more than 60 km to get to the hospital (0-20 km: n = 313, odds ratio [OR] 1; >20-40 km: n = 436, OR 1.09, 95% confidence interval [CI] 0.71 1.67; >40-60 km: n = 205, OR 0.97, 95%CI 0.57-1.65; >60 km: n = 79, OR 2.87, 95%CI 1.59-5.17). CONCLUSION: The mortality from tuberculosis was high, even amongst those living closest to the hospital, and did not rise significantly within 60 km. The situation may be different for the relatively small number of patients who come from further away. The distance travelled to hospital for initial diagnosis does not account for the relatively high mortality amongst tuberculosis patients in this area. PMID- 11931424 TI - Comparative analysis of partial structures of a peat humic and fulvic acid using one- and two-dimensional nuclear magnetic resonance spectroscopy. AB - Nuclear magnetic resonance (NMR) resonance integrals obtained from one dimensional NMR spectra provide semiquantitative contents of humic constituents with limited resolution in structural detail. When supplemented by connectivity information available from homo- and heteronuclear two-dimensional NMR spectra a more reliable assignment of humic substructures becomes available. This is demonstrated with a comparative one- and two-dimensional NMR analysis of a fulvic and a humic acid obtained from Eriophorum peat. An example of a detailed analysis of the proton chemical shift region normally attributed to carbohydrates shows substantial contributions from amino acids, amino and desoxy sugars, and highly oxidized aliphatic chains of intermediate length. The very good resolution of structural detail by a combined analysis of all NMR spectra shows that the effect of the fractionation procedure on the composition and chemical structure of humic materials is very significant. The comparison of the partial structures comprising humic acid (HA) and fulvic acid (FA) of the peat humic materials studied indicates that FA is diagenetically downstream of HA, favoring the biopolymer degradation (BD) model of humification. PMID- 11931425 TI - Improvements in the two-dimensional nuclear magnetic resonance spectroscopy of humic substances. AB - Understanding pollutant sorption, bioremediation of these pollutants, and their interactions with humic substances requires knowledge of molecular-level processes. New developments with nuclear magnetic resonance (NMR) experiments and labeled compounds have improved the overall understanding of these mechanisms. The advancements made with two-dimensional NMR show great promise, as structural information and hydrogen-carbon bond connectivity can be discerned. This communication presents the application of improved two-dimensional NMR methods, the double quantum filtered (DQF) correlation spectroscopy (COSY) and echo/anti echo heteronuclear single quantum coherence (HSQC) experiments, for use in structural studies of humic substances. Both experiments were found to produce significant improvements over the conventional COSY and heteronuclear multiple quantum coherence (HMQC) experiments that have been previously employed in similar studies. The more sensitive echo/anti-echo HSQC experiment produced more cross-peaks with higher resolution when compared with the HMQC spectra. The DQF COSY significantly suppressed the diagonal signals and allowed numerous signals previously hidden in the standard COSY experiment to be observed. These improvements will aid current characterization strategies of humic substances from soils, sediments, and water and their subsequent reactions with pollutants and microorganisms. PMID- 11931427 TI - Harvesting and climate effects on organic matter characteristics in British Columbia coastal forests. AB - As part of investigations into the effects of harvesting old-growth forest, we characterized carbon in five organic matter pools in eight forest chronosequences of coastal British Columbia. Each chronosequence comprised stands in four seral stages from regeneration (3-8 yr) to old-growth (>250 yr), with second-growth stands mostly of harvest origin. Stands were located in two biogeoclimatic subzones with contrasting climate (wetter, slightly cooler conditions on the west coast of Vancouver Island than on the east). Carbon concentrations in fine woody debris (FWD), forest floor (LFH), fine roots from LFH, and two water-floatable fractions from 10 to 30 cm mineral soil (MIN-ROOT, 2-8 mm and MIN-FLOAT, <2 mm) showed no significant effects due to climate, seral stage, or site. There were some significant differences in N concentrations, but none related to seral stage. Carbon-13 cross-polarization with magic-angle spinning (CPMAS) nuclear magnetic resonance (NMR) spectra with principal component analysis of relative areas also showed little harvesting effect, but greater variation related to input of coarse woody debris (CWD) vs. roots high in tannin. Overall, there tended to be more spectral features associated with wood and lignin in the west; whereas some MIN-ROOT samples from the drier east side had aromatic intensity attributed to charcoal. The minimal effects of one harvest on organic matter are most likely due to the large legacy effect; however, more intensive management will probably result in less CWD retention, less charcoal input, and less microsite variability in these pools of poorly decomposed organic matter. PMID- 11931426 TI - Solid-state carbon-13 nuclear magnetic resonance of humic acids at high magnetic field strengths. AB - Use of solid-state 13C nuclear magnetic resonance (NMR) spectroscopy has become commonplace in studies of humic substances in soils and sediments, but when modern high-field spectrometers are employed, care must be taken to ensure that the data obtained accurately reflect the chemical composition of these complex materials in environmental systems. In an effort to evaluate the quality of solid state 13C NMR spectra obtained with modern high-field spectrometers, we conducted a series of experiments to examine spectra of various humic acids taken under a variety of conditions. We evaluate conditions for obtaining semiquantitative cross polarization magic angle spinning (CPMAS) 13C NMR spectra of humic acids at high magnetic field and spinning frequency. We examine the cross polarization (CP) dynamics under both traditional and ramp CP conditions on Cedar Creek humic acid. Fitted equilibrium intensities from these CP dynamic studies compare to within 3.4% of the intensities determined from a Bloch decay spectrum of the same sample. With a 1-ms contact time, ramp CP and traditional CP spectra were acquired on this sample and were found to compare to within 5.4% of the Bloch decay spectrum; however, the signal-to-noise ratio per hour of data acquisition was found to double under ramp CP conditions. These results demonstrate the power of applying modern solid-state NMR techniques at high magnetic field strengths. With these techniques, high-quality, semiquantitative spectra can be quickly produced, allowing the application of solid-state NMR techniques to more environmentally relevant samples, especially those where the quantity is limited. PMID- 11931428 TI - Paramagnetic effects on solid state carbon-13 nuclear magnetic resonance spectra of soil organic matter. AB - The effects of paramagnetic species on solid state 13C nuclear magnetic resonance (NMR) spectra were quantified in a series of doping experiments. The degree of signal loss caused by paramagnetic metals was shown to depend not only on the quantity, but also on the nature of the paramagnetic species, as well as the intimacy of contact with the organic substrate and the type of NMR experiment. Two mechanisms of signal loss were distinguished--signal loss via loss of magnetic field homogeneity, which affects all 13C nuclei in a sample, and signal loss via interaction between electronic and nuclear spins, the effects of which were localized to the close environment of the paramagnetic species. Loss of field homogeneity is important for manganese species, but not for copper species, and is equally important for both cross polarization and Bloch decay experiments. The interaction between electronic and nuclear spins is highly dependent on the spin-lattice relaxation rate constant of the free electron (T1e), as cations with very short T1e values (e.g., Pr3+) cause less signal loss than cations with longer T1e values (e.g., Cu2+, Mn2+). Cross polarization spectra are shown to be more susceptible than Bloch decay spectra to this mechanism of signal loss. Signal loss and increased relaxation rates brought about by paramagnetic species can be used to provide information on soil organic matter (SOM) heterogeneity in the submicron range. This is demonstrated for SOM doped with paramagnetic cations where selective signal loss and increased relaxation rates are used to determine the nature of cation exchange sites. PMID- 11931429 TI - Multinuclear magnetic resonance analysis of two humic acid fractions from lowland rice soils. AB - To determine the effects of intensive cropping of tropical lowland rice (Oryza sativa L.) and the associated long-term soil submergence on chemical properties of soil organic matter, we used solid-state 13C and 15N and solution 31P nuclear magnetic resonance (NMR) spectroscopy to analyze the labile mobile humic acid (MHA) and the more recalcitrant calcium humate (CaHA) fractions extracted from a series of soils supporting several long-term field experiments in the Philippines. The soils varied mainly in degree of submergence and cropping intensity, ranging from a rainfed rice field without soil submergence to irrigated double- and triple-cropped fields in which soil remains submerged almost all year long. As reported previously, all analyses associated increasing intensity of rice cropping with larger proportions of less humified material in the MHA and CaHA, such as diester phosphorus (P), amide nitrogen (N), and phenolic carbon (C). We established significant correlations between proportions of various spectral areas as well as between some spectral areas and other humic acid (HA) properties such as visible light absorption and free radical concentration (positive indices of humification) and hydrogen (H) concentration (negative index of humification). For example, spectral proportions of heterocyclic N were positively, and proportions of amide N and phenolic C negatively, correlated with visible light absorption and free radical concentration, and each of these spectral proportions had an opposite sign when correlated with H concentration. The correlations of N-alkyl C proportions were the strongest with these properties and with other functional group proportions. PMID- 11931430 TI - Characterization of high-tannin fractions from humus by carbon-13 cross polarization and magic-angle spinning nuclear magnetic resonance. AB - Condensed tannins can be found in various parts of many plants. Unlike lignin there has been little study of their fate as they enter the soil organic matter pool and their influence on nutrient cycling, especially through their protein binding properties. We extracted and characterized tannin-rich fractions from humus collected in 1998 from a black spruce [Picea mariana (Mill.) Britton et al.] forest in Canada where a previous study (1995) showed high levels (3.8% by weight) of condensed tannins. A reference tannin purified from black spruce needles was characterized by solution 13C nuclear magnetic resonance (NMR) as a pure procyanidin with mainly cis stereochemistry and an average chain length of four to five units. The colorimetric proanthocyanidin (PA) assay, standardized against the black spruce tannin, showed that both extracted humus fractions had higher tannin contents than the original humus (2.84% and 11.17% vs. 0.08%), and accounted for 32% of humus tannin content. Consistent with the results from the chemical assay, the aqueous fraction showed higher tannin signals in the 13C cross-polarization and magic-angle spinning (CPMAS) NMR spectrum than the emulsified one. As both tannin-rich humus fractions were depleted in N and high in structures derived from lignin and cutin, they did not have properties consistent with recaldtrant tannin-protein complexes proposed as a mechanism for N sequestration in humus. Further studies are needed to establish if tannin protein structures in humus can be detected or isolated, or if tannins contribute to forest management problems observed in these ecosystems by binding to and slowing down the activity of soil enzymes. PMID- 11931431 TI - Enzymatic transformation and binding of labeled 2,4,6-trinitrotoluene to humic substances during an anaerobic/aerobic incubation. AB - Organic pollutants are degraded in soil and simultaneously nonextractable residues are formed. However, proof is lacking that this fixation has a detoxifying effect. We investigated the transformation and binding of 2,4,6 trinitrotoluene (TNT) with catechol or soil humic acid as cosubstrates. Carbon-14 labeled TNT and its reaction products were quantified by radiocounting; extractable compounds were identified by high performance liquid chromatography (HPLC). Bound and extractable residues of 15N-labeled TNT and metabolites were studied by 15N nuclear magnetic resonance spectroscopy (15N NMR). Since TNT is not easily transformed under oxidizing conditions an anaerobic/aerobic treatment was used. Anaerobic microorganisms from cow manure were used to reduce TNT during the anaerobic phase and subsequently, a laccase from Trametes villosa was used in the aerobic phase to oxidatively couple the metabolites to humic matter. Seventy four percent of TNT was immobilized with catechol as cosubstrate, but only 25% with humic acid. With catechol the main extractable component was TNT, while with humic acid it was mostly the metabolite 4-aminodinitrotoluene. For both co substrates, the spectra of immobilized metabolites obtained by solid-state 15N cross polarization magic angle spinning (CPMAS) NMR spectroscopy showed signals in the chemical shift region for protonated aromatic amino compounds. However, in the presence of catechol, an additional signal from nonextractable nitro groups was found, which could represent sequestered TNT. The partially reduced metabolites of TNT that formed nonextractable residues in humic acid are not likely to be remobilized easily and are thus regarded as detoxified. PMID- 11931432 TI - Formation of heteroaromatic nitrogen after prolonged humification of vascular plant remains as revealed by nuclear magnetic resonance spectroscopy. AB - In the search for the mechanisms involved in the immobilization of organic nitrogen in humified remains of vascular plants, the efforts of the present investigation were directed toward the examination of the transformation of nitrogenous compounds during the peat and coal stage by means of solid-state nuclear magnetic resonance (NMR) spectroscopy. While accumulation of heteroaromatic-N is not detected in most of the studied peat layers, a clear shoulder in the chemical shift region of pyrrole- or indole-N is observed in the solid-state 15N NMR spectrum of material from the deepest (and thus oldest) peat layer underlying the sapropel from Mangrove Lake, Bermuda (10000 years). This points to the assumption that transformation of nitrogen occurs between an advanced stage of peatification and an early stage of coalification. The observed sudden alteration in nitrogen functionality indicates that continuous accumulation of newly synthesized or selectively preserved biogenic structures is not responsible for the presence of heteroaromatic-N in these fossilized deposits. It seems rather likely that abiotic conditions, occurring during advanced sediment maturation, have an effect on the observed N transformation. With increasing coalification, pyrrole-type-N becomes the dominant form in the macromolecular coal network. Pyridine-type-N was only detected in a coal of anthracite rank. PMID- 11931433 TI - Analysis of potentially mobile phosphorus in arable soils using solid state nuclear magnetic resonance. AB - In many intensive agroecosystems continued inputs of phosphorus (P) over many years can significantly increase soil P concentrations and the risk of P loss to surface waters. For this study we used solid-state 31P nuclear magnetic resonance (NMR) spectroscopy, high-power decoupling with magic angle spinning (HPDec-MAS) NMR, and cross polarization with magic angle spinning (CP-MAS) NMR to determine the chemical nature of potentially mobile P associated with aluminum (Al) and calcium (Ca) in selected arable soils. Three soils with a range of bicarbonate extractable Olsen P concentrations (40-102 mg P kg(-1)) were obtained from a long term field experiment on continuous root crops at Rothamsted, UK, established in 1843 (sampled 1958). This soil has a threshold or change point at 59 mg Olsen P kg(-1), above which potentially mobile P (as determined by extraction with water or 0.01 M CaCl2) increases much more per unit increase in Olsen P than below this point. Results showed that CaCl2 and water preferentially extracted Al-P and Ca-P forms, respectively, from the soils. Comparison among the different soils also indicated that potentially mobile P above the threshold was largely present as a combination of soluble and loosely adsorbed (protonated-cross polarized) P forms largely associated with Ca, such as monetite (CaHPO4) and dicalcium phosphate dihydrate (CaHPO4-2H2O), and some Al-associated P as wavellite. The findings of this study demonstrate that solid-state NMR has the potential to provide accurate information on the chemical nature of soil P species and their potential mobility. PMID- 11931434 TI - Soil and litter phosphorus-31 nuclear magnetic resonance spectroscopy: extractants, metals, and phosphorus relaxation times. AB - Phosphorus-31 nuclear magnetic resonance (NMR) spectroscopy is an excellent tool with which to study soil organic P, allowing quantitative, comparative analysis of P forms. However, for 31P NMR to be tative, all peaks must be completely visible, and in their correct relative proportions. There must be no line broadening, and adequate delay times must be used to avoid saturation of peaks. The objective of this study was to examine the effects of extractants on delay times and peak saturation. Two samples (a forest litter and a mineral soil sample) and three extractants (0.25 M NaOH, NaOH plus Chelex (Bio-Rad Laboratories, Hercules, CA), and NaOH plus EDTA) were used to determine the differences in the concentration of P and cations solubilized by each extractant, and to measure spin-lattice (T1) relaxation times of P peaks in each extract. For both soil and litter, NaOH-Chelex extracted the lowest concentrations of P. For the litter sample, T1 values were short for all extractants due to the high Fe concentration remaining after extraction. For the soil sample, there were noticeable differences among the extractants. The NaOH-Chelex sample had less Fe and Mn remaining in solution after extraction than the other extractants, and the longest delay times used in the study, 6.4 s, were not long enough for quantitative analysis. Delay times of 1.5 to 2 s for the NaOH and NaOH-EDTA were adequate. Line broadening was highest in the NaOH extracts, which had the highest concentration of Fe. On the basis of these results, recommendations for future analyses of soil and litter samples by solution 31P NMR spectroscopy include: careful selection of an extractant; measurement of paramagnetic ions extracted with P; use of appropriate delay times and the minimum number of scans; and measurement of T1 values whenever possible. PMID- 11931435 TI - A novel technique for the pre-concentration and extraction of inositol hexakisphosphate from soil extracts with determination by phosphorus-31 nuclear magnetic resonance. AB - Inositol hexakisphosphate (IP6) is often the dominant form of soil organic phosphorus (P), but is rarely investigated because of the analytical difficulties encountered in its extraction, separation, and detection in environmental samples. In particular, recent advances in the study of soil organic P with 31P nuclear magnetic resonance (NMR) have been of limited use for the study of IP6, because the technique does not discriminate between IP6 and other forms of P. This was addressed by developing a novel analytical procedure using the retentive properties of gel-filtration gels for IP6, which allows the combined selective extraction and pre-concentration of IP6 from soil extracts with determination by 31P NMR. While the technique is still in the developmental stage, the results demonstrate that the gel does not interfere with 31P NMR analysis and retains IP6 to concentrations well above those required to give clear spectral signals. The technique has considerable potential for application to the study of IP6 in soil extracts and water samples and, with development, could help to answer fundamental questions regarding the dynamics of organic P in the environment. PMID- 11931436 TI - Flow and diffusion measurements in natural porous media using magnetic resonance imaging. AB - Flow and diffusion of water in natural porous media, quartz sand, and calcareous gravel were measured using a 1.5-T clinical magnetic resonance tomograph. The spatial resolution of the dynamic measurements was 1.32 x 1.32 x 5 mm3, and the time between two cross-sectional measurements was approximately 10 s. The measured coefficients of molecular diffusion for water were in good agreement with theoretical data. Flow was measured without any tracer at velocities between 0.15 and 6.67 mm/s. The results, based on a calibration within one part of the column, were in good agreement with data obtained from a tracer experiment and from a numerical model. It was possible to measure the flow velocity in larger pores and preferential flow paths directly. The results of the flow measurements in smaller pores reflected the mean velocity within that volume element. In that case the obtained values were close to the average linear velocity. Since the time resolution is high a monitoring of flow processes is possible. The pore space was imaged with a spatial resolution of 0.5 x 0.5 x 0.5 mm3. Here, the porosity of pores that are larger than 0.2 mm can be measured directly; for smaller pores a calibration is necessary. PMID- 11931437 TI - Nuclear magnetic resonance imaging for studies of flow and transport in porous media. AB - Nuclear magnetic resonance imaging (NMRI) methods for visualization of fluid flow and transport in porous media are reviewed in this paper. They are illustrated with experiments showing applications of velocity imaging, NMRI measurements of multiphase flow, and NMRI measurements of density flow. The latter two are compared with numerical simulations. The examples show the capacity of NMRI to give structural information both of the medium and the fluid distributions as well as their temporal development. The resulting data can be used in a black box white box comparison and as benchmarks for numerical models. PMID- 11931438 TI - Imaging of water flow in porous media by magnetic resonance imaging microscopy. AB - Magnetic resonance imaging (MRI) was used to study the flow of water in a column 14 mm in diameter packed with glass heads. The sample was fully saturated and water was pumped through the column using a peristaltic pump, at flow rates of 125 and 250 mL h(-1). This corresponds to mean velocities of 0.5 and 1 mm s(-1), given a porosity of 0.46 m3 m(-3). Nuclear magnetic resonance (NMR) images of the proton density and velocities within a 2-mm slice were taken at a spatial resolution of 0.15 x 0.15 x 2 mm3. At a mean pore water velocity of 1 mm s(-1) we approximated hydrodynamic dispersion using NMR-measured velocity distributions in a 2-mm slice through the sample. Additionally, we conducted a step pulse tracer experiment with chloride through the same column and at identical initial and boundary conditions. We fitted the convection-dispersion equation to the breakthrough curve and compared the column scale dispersion of the tracer experiment with the respective NMR estimate derived at the slice scale. PMID- 11931439 TI - Stray field nuclear magnetic resonance of soil water: development of a new, large probe and preliminary results. AB - Development, characterization, and preliminary results of a recent technique capable of local measurements of pore-size distribution by a spatially resolved low resolution nuclear magnetic resonance (NMR) technique are described. Potential environmental uses include studying the change in pore-size distribution caused by surface compaction, which influences surface runoff, and obtaining information on the physical state of non-aqueous compounds in porous materials, which should aid the selection of appropriate soil remediation methods. Stray field (STRAFI) imaging is an NMR technique that allows distortion free imaging of materials with short NMR relaxation times. The sample is placed in the strong axial fringe field gradient of a superconducting NMR magnet. We report on a new, unique, large 5-cm-diameter STRAFI probe, and its use for three preliminary test cases: water in ceramics of known pore size, paraffin wax and oil in sandstone rock, and water in soil at different matric potentials. The imaging is confined to one dimension with a spatial resolution of the order of 100 microm for protons. The optimum position for imaging occurs at 2.62 T and a gradient of 12.1 T/m. Water relaxation decay curves can be measured at any position in the 8-cm-long sample. These curves are decomposed into a series of terms each corresponding to a different pore size. Preliminary results show continuum fits to decay curves for a soil drained to three different matric potentials. Such information will be useful for interpreting water retention curves and will lead to understanding of the behavior of fluids in the vadose zone. PMID- 11931440 TI - Variations of permeability and pore size distribution of porous media with pressure. AB - Porosity and permeability of porous and fractured geological media decrease with the exploitation of formation fluids such as petroleum, natural gas, or ground water. This may result in ground subsidence and a decrease of recovery of petroleum, natural gas, or ground water. Therefore, an evaluation of the behavior of permeability and porosity under formation fluid pressure changes is important to petroleum and ground water industries. This study for the first time establishes a method, which allows for the measurement of permeability, porosity, and pore size distribution of cores simultaneously. From the observation of the pore size distribution by low-field nuclear magnetic resonance (NMR) relaxation time spectrometry the mechanisms of pressure-dependent porosity and permeability change can be derived. This information cannot be obtained by traditional methods. As the large-size pores or fractures contribute significantly to the permeability, their change consequently leads to a large permeability change. The contribution of fractures to permeability is even larger than that of pores. Thus, the permeability of the cores with fractures decreased more than that of cores without fractures during formation pressure decrease. Furthermore, it did not recover during formation pressure increase. It can be concluded that in fractures, mainly plastic deformation takes place, while matrix pores mainly show elastic deformation. Therefore, it is very important to keep an appropriate formation fluid pressure during the exploitation of ground water and petroleum in a fractured formation. PMID- 11931441 TI - Three-dimensional nickel ion transport through porous media using magnetic resonance imaging. AB - The transport of Ni2+ ions in a column, filled with porous media, was observed in three dimensions and time by magnetic resonance imaging (MRI) in a clinical scanner. For porous media we used glass beads or quartz sand in a saturated continuous flow mode. The magnetic moment of Ni2+ decreased the T1 relaxation time of 1H in aqueous solution. This concentration-dependent effect was used by a fast low angle shot (FLASH) MRI sequence for imaging the concentration of the dissolved ions. Since Ni2+ behaves as a conservative tracer under the chosen conditions, the tracer motion was representative for the water flow in the porous medium. Currently, we can achieve an isotropic spatial resolution of 1.5 mm and a temporal resolution of 170 s. The transport observation gives direct access to hydraulic flow properties of the porous media. The fluid flow velocity field was calculated by a fronttracking method and the statistical properties of the velocities were investigated. We also compared the experimental data with the three-dimensional particle tracking model PARTRACE, which uses the experimental flow field as input. PMID- 11931442 TI - Real-time, high-resolution quantitative measurement of multiple soil gas emissions: selected ion flow tube mass spectrometry. AB - A new technique is presented for the rapid, high-resolution identification and quantification of multiple trace gases above soils, at concentrations down to 0.01 microL L(-1) (10 ppb). The technique, selected ion flow tube mass spectrometry (SIFT-MS), utilizes chemical ionization reagent ions that react with trace gases but not with the major air components (N2, O2, Ar, CO2). This allows the real-time measurement of multiple trace gases without the need for preconcentration, trapping, or chromatographic separation. The technique is demonstrated by monitoring the emission of ammonia and nitric oxide, and the search for volatile organics, above containerized soil samples treated with synthetic cattle urine. In this model system, NH3 emissions peaked after 24 h at 2000 nmol m(-2) s(-1) and integrated to approximately 7% of the urea N applied, while NO emissions peaked about 25 d after urine addition at approximately 140 nmol m(-2) s(-1) and integrated to approximately 10% of the applied urea N. The monitoring of organics along with NH3 and NO was demonstrated in soils treated with synthetic urine, pyridine, and dimethylamine. No emission of volatile nitrogen organics from the urine treatments was observed at levels >0.01% of the applied nitrogen. The SIFT method allows the simultaneous in situ measurement of multiple gas components with a high spatial resolution of < 10 cm and time resolution <20 s. These capabilities allow, for example, identification of emission hotspots, and measurement of localized and rapid variations above agricultural and contaminated soils, as well as integrated emissions over longer periods. PMID- 11931444 TI - Industrial impact on marsh soils at the Bahia Blanca Ria, Argentina. AB - The Bahia Blanca Estuary is located in southern Buenos Aires province, Argentina. The area is linked to a petrochemical industrial complex, whose raw materials and final products contaminate the surrounding areas via atmospheric pollution and effluents, which are dumped in the estuary waters. To establish the effects of the industrial waste disposal on the nearest coastal soils, 17 samples were taken at different distances from the loading dock and the outfall pipes of the industrial complex. Later, the physicochemical characteristics of the soil samples, their hydrocarbon contents, sulfides, sulfates, Zn, Cu, and Pb were analyzed and a comparison was made to control samples, which were not affected by the industrial outfall. Hydrocarbons, Zn, Cu, and Pb contents were found at levels that modified the physical and chemical characteristics of the soils. The resistance to penetration shows that the thinner the film of water that surrounds the particles or aggregates, the smaller the migration of organic micelle, which settle on the surface of the contact material. This is demonstrated by the degree of cohesion reached by the particles and the strong influence on the index of hydrophobicity. The high porosity shows that the continuity of the porous space of the soil matrix is impeded by the presence of pollutants, which generate areas that are highly limiting to water flow. The oxidation-reduction potential and the low concentrations of soluble forms of Cu and Pb compared with their concentrations precipitated as sulfides confirm the action of the pollutants. PMID- 11931443 TI - Nitrous oxide and ammonia fluxes in a soybean field irrigated with swine effluent. AB - In the United States, swine (Sus scrofa) operations produce more than 14 Tg of manure each year. About 30% of this manure is stored in anaerobic lagoons before application to land. While land application of manure supplies nutrients for crop production, it may lead to gaseous emissions of ammonia (NH3) and nitrous oxide (N2O). Our objectives were to quantify gaseous fluxes of NH3 and N2O from effluent applications under field conditions. Three applications of swine effluent were applied to soybean [Glycine max (L.) Merr. 'Brim'] and gaseous fluxes were determined from gas concentration profiles and the flux-gradient gas transport technique. About 12% of ammonium (NH4-N) in the effluent was lost through drift or secondary volatilization of NH3 during irrigation. An additional 23% was volatilized within 48 h of application. Under conditions of low windspeed and with the wind blowing from the lagoon to the field, atmospheric concentrations of NH3 increased and the crop absorbed NH3 at the rate of 1.2 kg NH3 ha(-1) d(-1), which was 22 to 33% of the NH3 emitted from the lagoon during these periods. Nitrous oxide emissions were low before effluent applications (0.016 g N2O-N ha(-1) d(-1)) and increased to 25 to 38 g N2O-N ha(-1) d(-1) after irrigation. Total N2O emissions during the measurement period were 4.1 kg N2O-N ha(-1), which was about 1.5% of total N applied. The large losses of NH3 and N2O illustrate the difficulty of basing effluent irrigation schedules on N concentrations and that NH3 emissions can significantly contribute to N enrichment of the environment. PMID- 11931446 TI - Leaching from organic matter-rich soils by rain of different qualities: I. Concentrations. AB - Soil monoliths from an area exposed to acid precipitation and from an unpolluted area were used in a lysimeter experiment to study effects of different rain qualities on the chemical composition of the leachate from shallow soils rich in organic matter. The vegetation was either dominated by moorgrass [Molinia caerulea (L.) Moench] or heather [Calluna vulgaris (L.) Hull]. The lysimeters received either "acid rain" (pH 4.3) or "normal rain" (pH 5.3). High concentrations of dissolved organic carbon (DOC) were characteristic of the leachate. The different "rain" qualities had no significant influence on the DOC concentration. More DOC was, however, leached from lysimeters with heather vegetation. Roughly 50% of the aluminum (Al) was in complex with organic material and the Al charge was calculated to be between +1.4 and +2.0. Sulfate (SO4(2-)) was the only component that was significantly influenced by the treatment, as more was leached from lysimeters receiving "acid rain." Sulfate was poorly correlated with pH, suggesting that reduced SO4(2-) input would not necessarily lead to reduced acidity. Differences in the pH of the leachate due to the treatments were less than 0.15 pH units. Nitrate (NO3-) was only leached in very low concentrations and of little consequence for the leachate acidity. Some observations do, however, suggest that NO3- may contribute to acidification in episodes with high precipitation. High concentrations of Cl- in the leachate and a significant positive correlation between Cl-, H+, and base cations indicate that sea salt episodes may be important for soil acidification and acidity of the leachate. PMID- 11931447 TI - Chemistry of inorganic arsenic in soils: II. Effect of phosphorus, sodium, and calcium on arsenic sorption. AB - There are more than 10000 arsenic (As) contaminated sites in Australia. The ability of soils at these contaminated sites to sorb As is highly variable and appreciable amounts of As have been recorded in the subsurface soils. The potential risk of surface and ground water contamination by As at these sites is a major environmental concern. Factors that influence adsorption capacity of soils influence the bioavailability and subsequent mobility of As in soils. In the present study we investigated the effect of PO4(3-) and Na+ and Ca2+ on the sorption of AsV and AsIII by an Oxisol, a Vertisol, and two Alfisols. The presence of P (0.16 mmol L(-1)) greatly decreased AsV sorption by soils containing low amounts of Fe oxides (<100 mmol kg(-1)), indicating competitive adsorption between P and AsV for sorption sites. In contrast, the presence of a similar amount of P had little effect on the amount of AsV adsorbed by soils with high Fe content (>800 mmol kg(-1)). However, AsV sorption substantially decreased from 0.63 to 0.37 mmol kg(-1) as P concentration was increased from 0.16 to 3.2 mmol L(-1) in selected soils. This suggests increased competition between P and AsV for soil sorption sites, through either the higher affinity or the effect of mass action of the increasing concentration of P in solution. A similar effect of P on AsIII sorption was observed in the low sorbing Alfisol and high affinity Oxisol. However, the amount of AsIII sorbed by the Oxisol was much greater than the Alfisol for all treatments. The presence of Ca2+ increased the amount of AsV sorbed compared with that of Na+ and was manifested through changes in the surface charge characteristics of the soils. A similar trend in AsIII sorption was recorded with changes in index cation, although the effect was not as marked as recorded for AsV. PMID- 11931445 TI - Combination effect of light and toxicity in algal tests. AB - The sensitivity of Scenedesmus subspicatus against potassium dichromate is positively correlated to the photon flux density during the algal growth inhibition test. Low photon flux densities led to significantly reduced maximum effects and higher EC50 levels. To improve the testing of colored substances, we distinguished between the toxic effect (chemical part, represented by potassium dichromate) and the shading effect (physical part, simulated by reduced light intensities during the test) of a hypothetical light absorbing substance. The contribution of these single effects to the total inhibition varied greatly. At high concentrations of potassium dichromate (1.6 and 3.2 mg L(-1)) the physical part never exceeded 25% of the total inhibition, not even at strongest light reduction, while at low concentrations (0.2 and 0.4 mg L(-1)) the physical effect became more prominent when halving the amount of available light. Further, the combination effect of the chemical and the physical effect could be calculated well only by using the concept of independent action. Thus, if chemical and physical effects are measured in combination, as is the case in tests with dye stuffs, the current test protocol for the algal growth inhibition test may lead to incorrect estimations of the toxic potential. PMID- 11931449 TI - Dissolution kinetics of heavy metals in Dutch carbonate- and sulfide-rich freshwater sediments. AB - In two sulfide-rich freshwater sediments from the Biesbosch and Kromme Rijn River in the Netherlands differing in carbonate content and acid volatile sulfide (AVS) content, metal and sulfide dissolution kinetics were studied at different acid concentrations by varying both the procedure of acid addition and the extraction time. The establishment of equilibrium was monitored by measuring the pH in time, which reached a near constant value. The equilibrium pH was reached quickly when large amounts of acid were added and slowly when small amounts of acid were added. This observation was confirmed by the yield of extracted metals after either a 45-min or 24-h extraction over a pH range from 0 to 5. The pH factor seemed to be of more influence than time for the dissolution of metals. The amount of extracted metals was highly dependent on the metal itself due to its physico-chemical behavior. Although the sediments studied varied in carbonate content, acid volatile sulfide (AVS), and total metal content, the extracted fraction of metals compared with their total content in the sediment was similar for most metals. Finally, the AVS content as well as the ratio of simultaneously extracted metals (SEM; sum of Cd, Cu, Ni, Pb, and Zn) to AVS decreased with increasing pH. Because the SEM to AVS ratio may be used to set environmental quality criteria for the sediment compartment, this observation is of significance. PMID- 11931448 TI - In situ stabilization of soil lead using phosphorus and manganese oxide: influence of plant growth. AB - In situ stabilization of Pb contaminated soils can be accomplished by adding P and Mn(IV) oxide. However, the long-term efficacy of in situ stabilization under continual P removal through plant growth is unknown. Moreover, the effects these treatments have on phytoavailability of other metals (Cd and Zn) commonly associated with Pb in soil are not well understood. Greenhouse experiments using sudax [Sorghum vulgare (L.) Moench] and Swiss chard [Beta vulgaris (L.) Koch] were carried out to evaluate the effects of plant growth on soil Pb bioavailability to humans after addition of P and other amendments, and the effects of these treatments on Pb, Cd, and Zn phytoavailability in three metal contaminated soils. Eight treatments were used: zero P; 2500 mg of P as triple superphosphate (TSP); 5000 mg of P as TSP or phosphate rock (PR); 5000 mg of Mn oxide/kg; and combinations of Mn oxide and P as TSP or PR. The addition of P and/or Mn oxide significantly reduced bioavailable Pb, as measured by the physiologically based extraction test (PBET), in soils compared with the control even after extensive cropping. The PBET data also suggested that removal of P from soluble P sources by plants could negate the beneficial effects of P on bioavailable Pb, unless sufficient soluble P was added or soluble P was combined with Mn oxide. In general, Ph, Cd, and Zn concentrations in shoot tissues of sudax and Swiss chard were reduced significantly by TSP and did not change with the addition of PR. The combination of PR and Mn oxide significantly reduced Pb concentrations in plants compared with the control. PMID- 11931450 TI - Concentration, pH, and surface charge effects on cadmium and lead sorption in three tropical soils. AB - Reactions of heavy metals with soil are important in determining metal fates in the environment. Sorption characteristics of two heavy metals, Cd and Pb, in three tropical soils (Mollisol, Oxisol, and Ultisol) from Puerto Rico were assessed at varying metal concentrations (0 to 1.2 mM) and pH values (approximately 2 to 7). All soils sorbed more Pb than Cd. Sorption maxima were obtained for each metal for the Oxisol and Ultisol soils, but not the Mollisol. Sorption appeared to depend more on soil mineralogy than organic matter content. Sorption isotherms were linear within the sorption envelope with similar slopes for each soil-metal curve, when plotting metal sorption as a function of pH. Cadmium and Pb isotherms yielded average slopes of approximately 36+/-1 and 28+/ 1 units (percent increase in metal sorption per 1-unit increase in pH), respectively. Metal sorption depended more on metal type than soil composition. Cadmium sorption displayed a greater pH dependence than Pb. Cadmium sorption was less than or equal to the amount of negative surface charge except at pH values greater than the point of zero net charge (PZNC). This suggests that Cd was probably sorbed via electrostatic surface reactions and/or possible inner-sphere complexation at pH > 3.7. However, the amount of Pb sorbed by the Oxisol was greater than the amount of negative surface charge, suggesting that Pb participates in inner-sphere surface reactions. Lead was sorbed more strongly than Cd in our soils and poses less of a threat to underlying ground water systems due to its lower mobility and availability. PMID- 11931451 TI - Modeling phosphorus concentrations in Irish rivers using land use, soil type, and soil phosphorus data. AB - Modeling diffuse phosphorus (P) loss may indicate management strategies to minimize P loss from agricultural sources. An empirical model predicting flow weighted phosphorus concentrations (MRP) was derived using data collected from 35 Irish river catchments. Monitoring records of riverine P and stream flow data were used to calculate MRP values averaged for the years 1991-1994. These data were modeled using land use, soil type, and soil P data. Soil type in catchments was described using soil survey classifications weighted according to their P desorption properties from laboratory results. Soil test P concentrations for the studied watersheds were obtained from a national database. Soil P levels were weighted based on the results of field experiments measuring P losses in overland flow from fields at different soil test P levels. The 35 catchments were statistically clustered into two populations (A and B) based on differences in soil type, specifically, soil hydrology. Catchments in Cluster A had predominantly poorly drained soils and comparatively higher MRP concentrations (0.03-0.17 mg L(-1)) than Cluster B areas (0.01-0.7 mg L(-1)) with mostly well drained soils. Regression equations derived for A and B type catchments predicted MRP values with 68 and 62% of the variation explained in the models, respectively. Data extracted for the rest of the country were applied to the models to delineate areas at risk on a national scale. While the models were only moderately accurate they highlighted the influence of land management, specifically, high production grassland receiving high P inputs, in conjunction with the effect of soil type and soil hydrology on the transport of P to surface waters. PMID- 11931453 TI - Adsorption and degradation of the weak acid mesotrione in soil and environmental fate implications. AB - The ability of soils to adsorb and degrade pesticides strongly influences their environmental fate. This paper examines the adsorption and degradation of a weak acid, a new herbicide mesotrione 12-[4-(methylsulfonyl)-2-nitrobenzoyl]-1,3 cyclohexanedione], in 15 different soils from Europe and the USA. Experiments were conducted to understand the influence of soil properties, covering a wide range of soil textures, soil pH values (4.4 to 7.5), and organic carbon contents (0.6 to 3.35%). Mesotrione adsorption (Kd values ranged from 0.13 to 5.0 L/kg) was primarily related to soil pH, and to a lesser extent by percent organic carbon (%OC). As soil pH rose. mesotrione Kd values got smaller as mesotrione dissociated from the molecular to anionic form. Mesotrione degradation (half lives ranged from 4.5 to 32 d) was also related to soil pH, getting shorter as soil pH rose. Simple regression of mesotrione adsorption against soil pH and %OC and against degradation provided a close fit to the data. The correlation between mesotrione adsorption and degradation means that Kd and half-life values are only relevant for use in environmental fate assessment if these values are "paired" for the same soil pH and %OC. The implications were as illustrated for leaching, raising important issues about combining pesticide adsorption and degradation behavior in environmental fate assessments. PMID- 11931452 TI - Temporal and spatial variation of episodic wind erosion in unburned and burned semiarid shrubland. AB - Redistribution of soil, nutrients, and contaminants is often driven by wind erosion in semiarid shrublands. Wind erosion depends on wind velocity (particularly during episodic, high-velocity winds) and on vegetation, which is generally sparse and spatially heterogeneous in semiarid ecosystems. Further, the vegetation cover can be rapidly and greatly altered due to disturbances, particularly fire. Few studies, however, have evaluated key temporal and spatial components of wind erosion with respect to (i) erosion rates on the scale of weeks as a function of episodic high-velocity winds, (ii) rates at unburned and burned sites, and (iii) within-site spatial heterogeneity in erosion. Measuring wind erosion in unburned and recently burned Chihuahuan desert shrubland, we found (i) weekly wind erosion was related more to daily peak wind velocities than to daily average velocities as consistent with our findings of a threshold wind velocity at approximately 7 m s(-1); (ii) greater erodibility in burned vs. unburned shrubland as indicated by erosion thresholds, aerodynamic roughness, and nearground soil movement; and (iii) burned shrubland lost soil from intercanopy and especially canopy patches in contrast to unburned shrubland, where soil accumulated in canopy patches. Our results are among the first to quantify post fire wind erosion and highlight the importance of accounting for finer temporal and spatial variation in shrubland wind erosion. This finer-scale variation relates to semiarid land degradation, and is particularly relevant for predictions of contaminant resuspension and redistribution, both of which historically ignore finer-scale temporal and spatial variation in wind erosion. PMID- 11931454 TI - Molecular weight of dissolved organic matter-napropamide complex transported through soil columns. AB - Soil-derived dissolved organic matter (DOM) has been shown to form stable complexes with the herbicide napropamide [2-(alpha-naphthoxy-N,N diethylpropionamide] capable of enhancing the transport of napropamide through soil columns. Two soils, one containing sewage sludge-derived organic matter (SS) and the other having only natural organic matter (NoSS) were treated with napropamide and allowed to dry to promote complex formation. Soil columns were prepared by packing a 10-cm layer of untreated, dry, sieved soil followed by an overlying 5-cm layer of napropamide-treated soil. Columns were irrigated and the effluent collected and placed in dialysis chambers. After equilibration napropamide concentrations were determined on both sides of the membrane and complex and quantified based on the amount of napropamide unable to cross the membrane. it was found that for the SS soil 7% and for the NoSS 2.4% of the applied napropamide underwent facilitated transport. In addition, most of the complex transported through the columns had a molecular weight between 500 and 1000 Daltons (Da). The solutions from the SS soil were also found to have formed at least two distinct complexes that were resolved after passing through the untreated soil layer. The results obtained were in agreement with other published results and the techniques used offer a way to separate and concentrate DOM complexes from column effluents for further characterization. PMID- 11931455 TI - Rhizosphere acidification and cadmium uptake by strawberry clover. AB - Strawberry clover (Trifolium fragiferum L.) is a hardy legume found in indigenous or introduced populations throughout the world. Tolerance to saline and alkaline soils, flooding, and heavy metals make it a good prospect for reclamation projects. The research, described here, was conducted to: (i) characterize the morphological variation in plants from available seed sources, (ii) evaluate cadmium uptake and tolerances over a wide range of morphological variants, and (iii) elucidate the variability in the effects of roots on rhizosphere pH and the relationship to cadmium uptake. Seeds from selected accessions were planted in the greenhouse for comparison of morphological variation. The accessions examined had a mean height of 10.7+/-7 cm. Accessions 254916 and 237925 are tall with high rhizosphere pH values and might be useful in phytoremediation. Strawberry clover accessions were also grown hydroponically to examine differences in cadmium uptake. The ability of strawberry clover roots to change rhizosphere pH and take up cadmium was examined using culture tubes containing nutrient agar, a moderate level of cadmium, and a pH indicator dye. The results provided evidence for a negative correlation between rhizosphere pH and cadmium uptake. PMID- 11931456 TI - Simulating interactive effects of symbiotic nitrogen fixation, carbon dioxide elevation, and climatic change on legume growth. AB - The underlying mechanisms of interaction between the symbiotic nitrogen-fixation process and main physiological processes, such as assimilation, nutrient allocation, and structural growth, as well as effects of nitrogen fixation on plant responses to global change, are important and still open to more investigation. Appropriate models have not been adequately developed. A dynamic ecophysiological model was developed in this study for a legume plant [Glycine max (L.) Merr.] growing in northern China. The model synthesized symbiotic nitrogen fixation and the main physiological processes under variable atmospheric CO2 concentration and climatic conditions, and emphasized the interactive effects of these processes on seasonal biomass dynamics of the plant. Experimental measurements of ecophysiological quantities obtained in a CO2 enrichment experiment on soybean plants, were used to parameterize and validate the model. The results indicated that the model simulated the experiments with reasonable accuracy. The R2 values between simulations and observations are 0.94, 0.95, and 0.86 for total biomass, green biomass, and nodule biomass, respectively. The simulations for various combinations of atmospheric CO2 concentration, precipitation, and temperature, with or without nitrogen fixation, showed that increasing atmospheric CO2 concentration, precipitation, and efficiency of nitrogen fixation all have positive effects on biomass accumulation. On the other hand, an increased temperature induced lower rates of biomass accumulation under semi-arid conditions. In general, factors with positive effects on plant growth tended to promote each other in the simulation range, except the relationship between CO2 concentration and climatic factors. Because of the enhanced water use efficiency with a higher CO2 concentration, more significant effects of CO2 concentration were associated with a worse (dryer and warmer in this study) climate. PMID- 11931457 TI - Effects of arsenic concentrations and forms on arsenic uptake by the hyperaccumulator ladder brake. AB - Ladder brake (Pteris vittata L.) is a newly discovered arsenic hyperaccumulator. No information is available about arsenic effects on ladder brake. This study determined the effects of different arsenic concentrations (50 to 1000 mg kg(-1)) or forms (organic vs. inorganic and arsenite vs. arsenate) applied to soils on growth and arsenic uptake by ladder brake. Young plants were grown in a greenhouse for 12 or 18 wk. Ladder brake was highly tolerant of arsenic and survived in soil containing up to 500 mg As kg(-1). The fact that addition of arsenate up to 100 mg As kg(-1) increased fern biomass by 64 to 107%, coupled with higher arsenic concentration in younger fronds at low soil arsenic concentrations and older fronds at high soil arsenic concentrations, implies that arsenic may be beneficial for fern growth. Addition of 50 mg As kg(-1) was best for fern growth and arsenic accumulation, resulting in the highest fern biomass (3.9 g plant(-1)), bioconcentration factor (up to 63), and translocation factor (up to 25). With an exception of FeAsO4 and AlAsO4, which had the lowest effects due to their low solubility, little difference was observed among other arsenic forms mainly because of arsenic conversion in soil. Aboveground biomass was mostly responsible for accumulation of arsenic by plant (75-99%). Up to 26% of the added arsenic was removed by ladder brake, showing the high efficiency of ladder brake in arsenic removal. The results suggest that ladder brake may be a good candidate to remediate arsenic-contaminated soils. PMID- 11931458 TI - Jack pine growth and elemental composition are affected by saline tailings water. AB - In the processing of oil sands from Alberta's Athabasca formation, large quantities of alkaline, saline tailings and associated process-affected waters are produced. These waters may have a negative effect on plants used in reclamation of mined areas in this region of the northern boreal forest. In the present study, we examined the effects of process-affected water on the growth and elemental composition of jack pine (Pinus banksiana Lamb.) seedlings. Seedlings were grown in sand culture, and treated with tailings water to which mineral nutrients had been added. One-month-old seedlings were treated for 14 d, and all measured growth parameters were reduced. Growth and shoot elemental composition were also measured in seven-month-old seedlings that were treated for 10 wk with process-affected water. Shoots had significantly elevated levels of Na, Cl, S, P, B, and Sr, and significantly reduced levels of Fe, Mo, Ba, and K. The relationships between elemental composition and seedling growth and injury were examined using multiple regression. Growth rates, dry weights, and carotenoid content were reduced, but were not related to shoot elemental composition. Needle necrosis was positively related to tissue Na and Cl. Results indicate that reclamation planning must consider substrate Na and Cl levels when planting jack pine on tailings-affected sites. PMID- 11931459 TI - Pollutant removal efficacy of three wet detention ponds. AB - Monthly inflow and outflow data were collected from three wet detention ponds in Wilmington, North Carolina, for a 29-mo period. Two ponds drained urban areas consisting primarily of residential, mixed services, and retail usage, while the third mainly drained residential and golf course areas. One of the urban ponds achieved significant reductions in total nitrogen, nitrate, ammonium, total phosphorus, orthophosphate, and fecal coliform bacterial counts. This pond was characterized by a high length to width ratio, with most inputs directed into the upper area, and extensive coverage by a diverse community of aquatic macrophyte vegetation. The second urban pond achieved significant reductions in turbidity and fecal coliform bacterial counts, but there were no significant differences between inflowing and outflowing water nutrient concentrations. There were substantial suburban runoff inputs entering the mid- and lower-pond areas that short-circuited pollutant removal contact time. The golf course pond showed significant increases in nitrate, ammonium, total phosphorus, and orthophosphate in the outflow relative to the inflow, probably as a result of course fertilization. However, nutrient concentrations in the outflow water were low compared with discharges from a selection of other area golf courses, possibly a result of the outflow passing through a wooded wetland following pond discharge. To achieve good reduction in a variety of pollutants, wet pond design should include maximizing the contact time of inflowing water with rooted vegetation and organic sediments. This can be achieved through a physical pond design that provides a high length to width ratio, and planting of native macrophyte species. PMID- 11931460 TI - Fate and efficacy of polyacrylamide applied in furrow irrigation: full-advance and continuous treatments. AB - Polyacrylamide (PAM) is applied to 400000 irrigated hectares annually in the USA to control irrigation-induced erosion, yet the fate of dissolved PAM applied in irrigation water is not well documented. We determined the fate of PAM added to furrow streams under two treatments: Initial-10, 10 mg L(-1) PAM product applied only during the initial hours of the irrigation, and Cont-1, 1.0 mg L(-1) PAM product applied continuously during the entire irrigation. The study measured PAM concentrations in 167-m-long PAM-treated furrow streams and along a 530-m tail ditch that received this runoff. Soil was Portneuf silt loam (coarse-silty, mixed, superactive, mesic Durinodic Xeric Haplocalcid) with 1.5% slope. Samples were taken at three times during the irrigations, both during and after PAM application. Polyacrylamide was adsorbed to soil and removed from solution as the streams traversed the soil-lined channels. The removal rate increased with stream sediment concentration. Stream sediment concentrations were higher when PAM concentrations were <2 mg L(-1) a.i., for early irrigations, and when untreated tributary flows combined with the stream. In these cases, PAM concentration decreased to undetectable levels over the flow lengths used in this study. When inflows contained >6 mg L(-1) PAM a.i., stream sediment concentrations were minimal and PAM concentrations did not change down the furrow, though they decreased to undetectable levels within 0.5 h after application ceased. One percent of applied PAM was lost in tail-ditch runoff. This loss could have been eliminated by treating only the furrow advance or not treating the last two irrigations. PMID- 11931462 TI - Phosphorus leaching from biosolids-amended sandy soils. AB - Increasing emphasis on phosphorus (P)-based nutrient management underscores the need to understand P behavior in soils amended with biosolids and manures. Laboratory and greenhouse column studies characterized P forms and leachability of eight biosolids products, chicken manure (CM), and commercial fertilizer (triple superphosphate, TSP). Bahiagrass (Paspalum notatum Flugge) was grown for 4 mo on two acid, P-deficient Florida sands, representing both moderate (Candler series: hyperthermic, uncoated Typic Quartzipsamments) and very low (Immokalee series: sandy, siliceous, hyperthermic Arenic Alaquods) P-sorbing capacities. Amendments were applied at 56 and 224 kg P(T) ha(-1), simulating P-based and N based nutrient loadings, respectively. Column leachate P was dominantly inorganic and lower for biosolids P sources than TSP. For Candler soil, only TSP at the high P rate exhibited P leaching statistically greater (alpha = 0.05) than control (soil-only) columns. For the high P rate and low P-sorbing Immokalee soil, TSP and CM leached 21 and 3.0% of applied P, respectively. Leachate P for six biosolids was <1.0% of applied P and not statistically different from controls. Largo biosolids, generated from a biological P removal process, exhibited significantly greater leachate P in both cake and pelletized forms (11 and 2.5% of applied P, respectively) than other biosolids. Biosolids P leaching was correlated to the phosphorus saturation index (PSI = [Pox]/[Al(ox) + Fe(ox)]) based on oxalate extraction of the pre-applied biosolids. For hiosolids with PSI < or = approximately 1.1, no appreciable leaching occurred. Only Largo cake (PSI = 1.4) and pellets (PSI = 1.3) exhibited P leaching losses statistically greater than controls. The biosolids PSI appears useful for identifying biosolids with potential to enrich drainage P when applied to low P-sorbing soils. PMID- 11931461 TI - Fate of nitrate and bromide in an unsaturated zone of a sandy soil under citrus production. AB - Understanding water and nutrient transport through the soil profile is important for efficient irrigation and nutrient management to minimize excess nutrient leaching below the rootzone. We applied four rates of N (28, 56, 84, and 112 kg N ha(-1); equivalent to one-fourth of annual N rates being evaluated in this study for bearing citrus trees), and 80 kg Br- ha(-1) to a sandy Entisol with >25-yr old citrus trees to (i) determine the temporal changes in NO3-N and Br- distribution down the soil profile (2.4 m), and (ii) evaluate the measured concentrations of NO3-N and Br- at various depths with those predicted by the Leaching Estimation and Chemistry Model (LEACHM). Nitrate N and Br concentrations approached the background levels by 42 and 214 d, respectively. Model-predicted volumetric water content and concentrations of NO3-N and Br- at various depths within the entire soil profile were very close to measured values. The LEACHM data showed that 21 to 36% of applied fertilizer N leached below the root zone, while tree uptake accounted for 40 to 53%. Results of this study enhance our understanding of N dynamics in these sandy soils, and provide better evaluation of N and irrigation management to improve uptake efficiency, reduce N losses, and minimize the risk of ground water nitrate contamination from soils highly vulnerable to nutrient leaching. PMID- 11931464 TI - A time-saving method for higher plant tests in hydroculture. AB - For higher plant tests in hydroculture we developed a method to unify the usually separately performed germination and growth testing. This method renders unnecessary the time-consuming and laborious installation of the germinated plants into the growth system. PMID- 11931463 TI - Microcosm wetlands for wastewater treatment with different hydraulic loading rates and macrophytes. AB - Constructed wetlands (CW) usually require large land areas for treating wastewater. This study evaluated the feasibility of applying CW with less land requirement by operating a group of microcosm wetlands at a hydraulic retention time (HRT) of less than 4 d in southern Taiwan. An artificial wastewater, simulating municipal wastewater containing 200 mg L(-1) of chemical oxygen demand (COD), 20 mg L(-1) of NH4+-N (AN), and 20 mg L(-1) of PO4(3-)-P (OP), was the inflow source. Three emergent plants [reed, Phragmites australis (Cav.) Trin. ex Steud.; water primrose, Ludwigia octovalvis (Jacq.) P.H. Raven; and dayflower, Commelina communis L.] and two floating plants [water spinach, Ipomoea aquatica Forssk.; and water lettuce, Pistia stratiotes L.] plants were tested. The planted systems showed more nutrient removal than unplanted systems; however, the type of macrophytes in CW did not make a major difference in treatment. At the HRTs of 2 to 4 d, the planted system maintained greater than 72,80, and 46% removal for COD, AN, and OP, respectively. For AN and OP removal, the highest efficiencies occurred at the HRT of 3 d, whereas maximum removal rates for AN and OP occurred at the HRT of 2 d. Both removal rates and efficiencies were reduced drastically at the HRT of 1 d. Removals of COD, OP, and AN followed first-order reactions within the HRTs of 1 to 4 d. The efficient removals of these constituents obtained with HRT between 2 and 4 d indicated the possibility of using a CW system for wastewater treatment with less land requirement. PMID- 11931465 TI - Characterization of a high-dose-rate 90Sr-90Y source for intravascular brachytherapy by using the Monte Carlo code PENELOPE. AB - Radiation treatment with catheter-based beta-emitter sources is currently under clinical trial to prevent restenosis. In the present paper, we address the characterization of the high-dose-rate 90Sr-90Y seeds of the Beta-Cath system supplied by Novoste Corporation, one of the commercially available sources for intravascular brachytherapy. The Monte Carlo code PENELOPE has been used to simulate the transport of electrons emitted by the encapsulated 90Sr-90Y seeds. The calculated radial dose function and anisotropy function for a single seed in water are compared with simulation results from other authors. Regarding g(r), the present result lies between the ITS3 and EGS4 curves, being somewhat closer to ITS3, while in the case of F(r, theta) some differences appear for certain angular intervals and radial distances. In order to put the observed differences into perspective, we have calculated radial doses for point isotropic sources in water. Our results for 0.5 and 1 MeV electrons are in good agreement with simulations using EGSnrc, and an excellent agreement is obtained with ITS for point 90Sr-90Y emitters. Dose distributions in water are calculated for source 'trains' consisting of 1, 2, 3, 4, 5, 9 and 12 seeds. The dose at the source midplane is enhanced if the number of seeds is up to 4, and saturates for trains with 5 or more seeds. We also compare the dose distribution obtained by simply adding the contributions of individual seeds with the simulation of the complete source train. It is found that both calculation procedures yield essentially the same result for distances greater than about 2 mm. Finally, the contribution of bremsstrahlung photons to the dose is briefly analysed. PMID- 11931467 TI - Optimal flattening filter shape of a surface brachytherapy applicator. AB - As an alternative to standard treatment of superficial lesions with surgery and/or superficial/orthovoltage x-ray irradiation, radioactive moulds can be used. However, close proximity of the source to the treatment region leads to significant dose inhomogeneities in the tissue. Analytical and fast numerical methods have been developed that can calculate a corresponding flattening filter that evens out the dose distribution at the surface. Monte Carlo transport calculations were used to enable accurate treatment of the transport through the geometry. It was found that the use of pre-calculated transport parameters like dependence of the attenuation coefficient and scattering with depth is required for fast calculation of the flattening filter shape. Only 1-2 iterations were needed for a successful convergence to the filter shape that flattens out the dose distribution at the surface to within 1%. The developed methods are very general and could be applied with some minor modifications to other problems, where shaping of the irradiation field is required. PMID- 11931466 TI - A flattening filter for brachytherapy skin irradiation. AB - Radioactive sources in close contact offer an alternative to superficial radiation in the treatment of skin lesions. A flattening filter was designed for a lead surface applicator to improve the skin dose distribution of a high dose rate (HDR) brachytherapy unit (Nucletron). At three heights from the opening (10, 15 and 25 mm) of the cylindrical applicator, the 192Ir source can be driven into the centre of the applicator. Thin sheets of lead foil (0.2 mm) were cut into circular shapes and placed in the opening to build a cylindrical cone that acts as a flattening filter. The shape of the cone was optimized in an iterative process using a spreadsheet and the resulting dose distribution under the applicator was determined using radiosensitive film. The use of the filter improved the dose distribution in a plane perpendicular to the beam axis to be within +/- 5% of the central axis dose. The present applicator and flattening filter together with an HDR brachytherapy unit offer an alternative for skin irradiation where a superficial unit is not available or will be replaced with a more flexible device. As the depth dose characteristics can be modified using different source-to-surface distances, the dose throughout the patient's skin can be shaped as desired by the radiation oncologist using a compensator design type approach. PMID- 11931468 TI - Non-linear model for the kinetics of 10B in blood after BPA-fructose complex infusion. AB - A numerical model with a memory effect was created to describe the kinetics of 10B in blood after a single 4-dihydroxyborylphenylalanine-fructose complex (BPA F) infusion in boron neutron capture therapy (BNCT). The model formulation was based on the averaged data from 10 glioma patients from the Brookhaven National Laboratory (BNL) BNCT-trials. These patients received a 2 h i.v. infusion of a BPA-fructose complex that delivered 290 mg BPA/kg body weight. The model was validated by fitting the original BNL patient data and new patient data from the Finnish BNCT-trials. The new 3-parameter non-linear model provided mean absolute differences between the measured and estimated 10B concentrations in blood that were less than 3.9% when used to simulate actual patient irradiations that comprised two irradiation fields separated by a break to reposition the patient. The flexibility of the model was successfully tested with two different infusion protocols. The patient data were modelled with a two-compartment model and a bi exponential fit for comparison. The 3-parameter model is better than previously described models in predicting the time course of blood 10B concentration after cessation of intravenous infusion of BPA-fructose. PMID- 11931469 TI - Nuclear interactions in proton therapy: dose and relative biological effect distributions originating from primary and secondary particles. AB - The dose distribution delivered in charged particle therapy is due to both primary and secondary particles. The secondaries, originating from non-elastic nuclear interactions, are of interest for three reasons. First, if fast Monte Carlo treatment planning is envisaged, the question arises whether all nuclear interaction products deliver a significant contribution to the total dose and, hence, need to be tracked. Second, there could be an enhanced relative biological effectiveness (RBE) due to low energy and/or heavy secondaries. Third, neutrons originating from nuclear interactions may deliver dose outside the target volume. The particle yield from different nuclear interaction channels as a function of proton penetration depth was studied theoretically for different proton beam energies. Three-dimensional dose distributions from primary and secondary particles were simulated for an unmodulated 160 MeV proton beam with and without including a slice of bone material and for a spread-out Bragg peak (SOBP) of 3 x 3 x 3 cm3 in water. Secondary protons deliver up to 10% of the total dose proximal to the Bragg peak of an unmodulated proton beam and they affect the flatness of the SOBP. Furthermore, they cause a dose build-up due to forward emission of secondary particles from nuclear interactions. The dose deposited by d, t, 3He and alpha-particles was found to contribute less than 0.1% of the total dose. The dose distal to the target volume caused by liberated neutrons was studied for four proton beam energies in the range of 160-250 MeV and found to be below 0.05% (2 cm distal to SOBP) of the prescribed target dose for a 3 x 3 x 3 cm3 target. RBE values relative to 60Co were calculated proximal to and within the SOBP. The RBE proximal to the Bragg peak (100% dose) is influenced by secondary particles (mainly protons and a-particles) with a strong dose dependency resulting in RBE values up to 1.2 (2 Gy; inactivation of V79). Depending on the endpoint considered, secondary particles cause a shift in RBE by up to 8% at 2 Gy. In contrast, the RBE in the Bragg peak is almost entirely determined by primary protons due to a decreasing secondary particle fluence with depth. RBE values up to 1.3 (2 Gy; inactivation of V79) at 1 cm distal to the Bragg peak maximum were found. The inactivations of human skin fibroblasts and mouse lymphoma cells were also analysed and reveal a substantial tissue dependency of the total RBE. The outcome of this study shows that elevated RBE values occur not only at the distal edge of the SOBP. Although the variations are modest, and in most cases might have no observable clinical effect, they might have to be considered in certain treatment situations. The biological effect downstream of the target caused by neutrons was analysed using a radiation quality factor of 10. The biological dose was found to be below 0.5% of the prescribed target dose (for a 3 x 3 x 3 cm3 SOBP) but depends on the size of the SOBP. This dose should not be significant with respect to late effects, e.g. cancer induction. PMID- 11931470 TI - Beam configurations for 3D tomographic intensity modulated radiation therapy. AB - We extend the theory of three-dimensional (3D) tomographic intensity modulated radiation therapy (IMRT). The geometry consists of two-dimensional modulated beams on a sphere centred in the tumour. The theory provides an efficient algorithm for computing beam modulation patterns that approximately 'reconstruct' the prescribed dose function. In this paper optimum beam numbers are estimated from dose function spherical harmonics using the 3D projection-slice theorem. An extension to three dimensions of the 'Bow Tie' criterion for beam numbers is derived. The effects of insufficient beam front sampling and beam numbers are characterized with a configuration-dependent matrix. Factors that independently increase beam numbers, such as tumour size and shape, are related to the spherical harmonic content in the dose function. Examples of tomographic IMRT reconstruction with a 3D concave tumour are given. PMID- 11931471 TI - Investigation into the relationship between body surface area and total body potassium using Monte Carlo and measurement. AB - The use of body surface area (BSA) as a means of indexing chemotherapy doses is widespread even though the value of this practice is uncertain. In principle, the body cell mass (BCM) more closely represents the body's metabolic size and this is investigated here as an alternative to BSA; since 98% of body potassium is intracellular the derivation of total body potassium (TBK) via the measurement of 40K in a whole body counter (WBC) will provide a useful normalizing index for metabolic size, potentially avoiding toxicity and underdosing. The Queen Elizabeth Hospital WBC has been used in this study, initially involving single geometrical phantoms and then combinations of these to simulate human body habitus. Monte Carlo N-particle (MCNP) codes were constructed to model the phantoms and simulate the measurements made in the WBC. Efficiency corrections were derived by comparing measurement and modelled data for each detector separately. A method of modelling a person in the WBC as a series of ellipsoids was developed. Twenty-four normal males and 24 females were measured for their 40K emissions. Individual MCNP codes were constructed for each volunteer and the results used in conjunction with the measurements to derive TBK, correcting for body habitus effects and detector efficiencies. An estimate of the component of error arising from sources other than counting statistics was included by analysing data from the measurement of phantoms. The total residual errors (expressed as coefficients of variation) for males and females were 10.1% and 8.5% respectively. The measurement components were determined to be 2.4% and 2.5%, implying that the biological components were 9.8% and 8.1% respectively. These results suggest that the use of BSA for indexing chemotherapy doses is likely to give rise to clinically significant under- or overdosing. PMID- 11931472 TI - The role of de-excitation electrons in measurements with graphite extrapolation chambers. AB - A method is described for determining the absorbed dose to graphite formedium energy x-rays (50-300 kV). The experimental arrangement consists of an extrapolation chamber which is part of a cylindrical graphite phantom of 30 cm diameter and 13 cm depth. The method presented is an extension of the so-called two-component model. In this model the absorbed dose to graphite is derived from the absorbed dose to the air of the cavity formed by the measuring volume. Considering separately the contributions of the absorbed dose to air in the cavity from electrons produced in Compton and photoelectric interactions this dose can be converted to the absorbed dose to graphite in the limit of zero plate separation. The extension of the two-component model proposed in this paper consists of taking into account the energy transferred to de-excitation electrons, i.e. Auger electrons, which are produced as a consequence of a photoelectric interaction or a Compton scattering process. For the system considered, these electrons have energies in the range between about 200 eV and 3 keV and hence a range in air at atmospheric pressure of 0.2 mm or less. As the amount of energy transferred to the de-excitation electrons is different per unit mass in air and in graphite, there is a region, about 0.2 mm thick, of disturbed electronic equilibrium at the graphite-to-air interface. By means of the extension proposed, the x-ray tube voltage range over which a graphite extrapolation chamber can be used is lowered from 100 kV in the case of the two component model down to at least 50 kV. PMID- 11931473 TI - Automatic MR volume registration and its evaluation for the pelvis and prostate. AB - A three-dimensional (3D) mutual information registration method was created and used to register MRI volumes of the pelvis and prostate. It had special features to improve robustness. First, it used a multi-resolution approach and performed registration from low to high resolution. Second, it used two similarity measures, correlation coefficient at lower resolutions and mutual information at full resolution, because of their particular advantages. Third, we created a method to avoid local minima by restarting the registration with randomly perturbed parameters. The criterion for restarting was a correlation coefficient below an empirically determined threshold. Experiments determined the accuracy of registration under conditions found in potential applications in prostate cancer diagnosis, staging, treatment and interventional MRI (iMRI) guided therapies. Images were acquired in the diagnostic (supine) and treatment position (supine with legs raised). Images were also acquired as a function of bladder filling and the time interval between imaging sessions. Overall studies on three patients and three healthy volunteers, when both volumes in a pair were obtained in the diagnostic position under comparable conditions, bony landmarks and prostate 3D centroids were aligned within 1.6 +/- 0.2 mm and 1.4 +/- 0.2 mm, respectively, values only slightly larger than a voxel. Analysis suggests that actual errors are smaller because of the uncertainty in landmark localization and prostate segmentation. Between the diagnostic and treatment positions, bony landmarks continued to register well, but prostate centroids moved towards the posterior 2.8-3.4 mm. Manual cropping to remove voxels in the legs was necessary to register these images. In conclusion, automatic, rigid body registration is probably sufficiently accurate for many applications in prostate cancer. For potential iMRI-guided treatments, the small prostate displacement between the diagnostic and treatment positions can probably be avoided by acquiring volumes in similar positions and by reducing bladder and rectal volumes. PMID- 11931474 TI - Simultaneity of foetal heart rate acceleration and foetal trunk movement determined by foetal magnetocardiogram actocardiography. AB - We describe a new method of actocardiography, foetal magnetocardiogram (fMCG) actocardiography, which is based on the high sensitivity of the fMCG to foetal trunk movements. We demonstrate the efficacy of the method by applying it to assess the simultaneity of onset of foetal heart rate (FHR) acceleration and foetal trunk movement. The analysis was restricted to events for which the onset of FHR accelerations and foetal movements could be determined accurately, i.e. when FHR was stable and near the average quiescent level just prior to the acceleration. We found that FHR accelerations coincided with or preceded foetal movements nearly all the time. This supports the hypothesis of coordinated control of FHR accelerations and foetal movements more strongly than prior studies, based on other techniques. We also found that beat-to-beat FHR variability often decreased at or near the start of FHR accelerations and that this occurrence was an accurate marker of foetal movement onset, even when foetal movement onset lagged FHR accelerations. PMID- 11931476 TI - Biologically effective uniform dose: an appropriate quantity to specify and report radiotherapy treatment plans? PMID- 11931475 TI - Speed of particles ejected from animal skin by CO2 laser pulses, measured by laser Doppler velocimetry. AB - During ablation of tissue with laser pulses rapid sublimation of matter occurs and high pressures are exerted within the tissue, resulting in steam. smoke and particles being expelled. In this paper we report the speed of particles ejected from animal tissue exposed to CO2 laser pulses measured directly by laser Doppler velocimetry (LDV). Speeds recorded just above animal skin were in the range of 9 to 18 m s(-1) for laser pulses of 128 to 384 J cm(-2) respectively. Aerodynamic turbulence slowed the particles down to a critical speed Vc of 4 m s(-1) within a few millimetres above the laser ablation site. Once the particles reach this minimum speed, if no collisions occur, they will only decelerate by gravitational action and the residual kinetic energy will send the particles up to about 0.87 m from the skin surface. Since ejected particles may carry viable cells, acting as disease vectors during laser surgery, our results suggest that the LDV technique should be used to measure the speed of particles ejected from healthy or pathological human tissues, helping to establish safe conditions during laser surgery. PMID- 11931477 TI - D, an effective uniform dose linked to the probability of response. PMID- 11931478 TI - Professional responsibility. PMID- 11931479 TI - Misfit crown. PMID- 11931480 TI - Light cure units. PMID- 11931481 TI - Occlusal misconceptions. PMID- 11931482 TI - Better opportunities for women dentists: a review of the contribution of women dentists to the workforce. AB - In June 2000 the Department of Health commissioned a review to examine the need for improvements to the employment opportunities for women dentists in the National Health Service (NHS) across England. Dame Margaret Seward carried out the review, which was published in September 2001. The review was considered necessary for four main reasons. Firstly, workforce panning, because now more than 50% of new entrants to dental undergraduate courses in the UK are female and by 2020 over 50% of all practising dentists will be female. Secondly, evidence that 50% of women in dentistry work for no more than two days per week for the NHS. Thirdly, most women work either as associates in general dental practice (GDP) or in the Community Dental Service (CDS). Lastly, the perception that women find it difficult to return to dentistry after taking a career break. PMID- 11931483 TI - Crowns and extra-coronal restorations: materials selection. AB - Materials selection is the second in the series on crowns and other extra-coronal restorations. Some of us are less than inspired by dental materials science. Nevertheless, many of the things that concern us clinically with crowns and their alternatives are based on material properties. We worry about the strength of the restoration, how well it fits and its aesthetics. We also worry about wear, occlusal control and biocompatibility. Not least of our concerns are dental laboratory charges, which inevitably have to be passed on to the patient. PMID- 11931485 TI - Referral for secondary restorative dental care in rural and urban areas of Scotland: findings from the Highlands Et Islands Teledentistry Project. AB - OBJECTIVES: To compare the reported level of use of secondary care services for restorative dental care in rural and urban areas of Scotland. DESIGN: Postal questionnaire survey SUBJECTS AND METHODS: Postal questionnaire sent to all dentists in the Highland region, the island regions in Scotland and Dumfries Et Galloway (n = 150) and an equal number were sampled from the remainder of Scotland stratified by health board area. Non-respondents were sent 2 reminders after which 62% of the sample had responded. RESULTS: Most dentists (85%) who practised in what they considered were urban areas of Scotland said they felt that they had good access to a secondary referral service. Whereas most of those who practised in what they considered were rural areas either said they had no access to such a service (26%) or that access was difficult (53%), only 3% of those in urban areas said they had no access to a secondary restorative consultative service compared with 14% of dentists practising in rural areas of mainland Scotland and 54% of those practising on Scottish islands. CONCLUSIONS: The survey suggests the people of the Scottish islands and some of the remoter parts of the Scottish mainland would be among those who might benefit from improvement in access to a restorative dentistry consultant service. PMID- 11931484 TI - The fate of the carious primary teeth of children who regularly attend the general dental service. AB - OBJECTIVE: To describe the care and resultant outcomes of the carious primary teeth of children who regularly attend the General Dental Service (GDS). SETTING: Four districts in the North West of England SUBJECTS AND MATERIALS: A retrospective study of the case notes of 677 children who received their dental care from 50 general dental practitioners (GDPs). Each dentist must have had a minimum of 10 patients and a maximum of 20 patients whose care had been provided by the same dentist from or before the age of five to the age of 14. All of the children included in the study had a history of approximal caries. The outcomes of interest were extraction due to pain or sepsis, or exfoliation and whether or not a tooth had given rise to the prescription of a course of antibiotics. Teeth that did not have a history of extraction were assumed to have exfoliated naturally. Logistic regression models, taking into account the clustering of the teeth within patients, were fitted to compare the outcomes for restored and unrestored teeth according to size of lesion (one or two surface), age caries was first recorded and by tooth type. RESULTS: A total of 4,056 teeth had been either recorded as carious or had received an intervention of some kind. Some 44.1% (N=1,789) of these teeth were extracted, however only 475 (11.7%) were extracted due to pain or sepsis. Of the teeth with a documented history of caries or restoration and for which an outcome was recorded (N=3,145), most first (81.1%) and second (84.3%) carious primary molars were filled during their lifetime, but only 40.5% of primary carious anterior teeth were filled. The majority of carious primary teeth exfoliated naturally. There was no difference in the proportions of teeth extracted due to pain or sepsis whether a carious tooth was restored or left unrestored, either by cavity type or by tooth type, after controlling for age when caries was first recorded. There was also no difference in the number of filled or unfilled carious teeth that caused a course of antibiotics to be prescribed. CONCLUSIONS: Treatment by extraction was common, but GDPs restored the majority of carious primary molar teeth of their regularly attending child patients. The bulk of carious teeth exfoliated naturally irrespective of whether they were filled or not. The reasons for these findings require further investigation. PMID- 11931486 TI - Smoking cessation strategies and periodontal disease in young adults. PMID- 11931487 TI - Focus awards 2001. PMID- 11931488 TI - Passion for surgical research. PMID- 11931489 TI - Arterio-venous CO2 removal (AVCO2R) perioperative management: rapid recovery and enhanced survival. AB - Percutaneous arteriovenous CO2 removal (AVCO2R) uses a simple arteriovenous (A-V) shunt for near-total CO2 removal that allows significant reductions in minute ventilation. We critically reviewed our algorithm-directed perioperative anesthesia management in our LD40 ovine smoke-burn injury model of acute respiratory distress syndrome (ARDS) treated with AVCO2R. General anesthesia is required for: (1) Vascular access followed by ARDS model development by smoke insufflation (36 breaths) plus 40% TBSA III degrees burn with mechanical ventilation. Induction: 12.5 mg/kg im ketamine and 4% halothane by mask, then intubation. Maintenance: 1.0-2.5% halothane in 100% O2; (2) When PaO2/FiO2 < 200 (48-52 h), sheep randomized to the AVCO2R (n = 8) or SHAM (n = 8) procedure. Induction: 66% N2O and 5% isoflurane in balance O2. Maintenance: 1.5-2.5% isoflurane in 100% O2 for AVCO2R, cannulation (10F carotid artery, 14F jugular vein); (3) Postop, both groups had algorithm-directed ventilator management, identical heparin (ACT > 300 s), fluid, and analgesia management. All sheep met criteria for ARDS, survived anesthesia, and were standing by 0.5-5 h. There were no complications attributable to anesthesia. The absence of anesthesia-related complications allows model development for outcomes studies for ARDS in general and AVCO2R specifically. PMID- 11931490 TI - Influence of early drainage of intraperitoneal phospholipids on efficacy of adhesion prevention. AB - Postoperative peritoneal adhesions impose a long-term risk of morbidity and mortality. Adjunctive means are needed to prevent these complications. In previous studies we could demonstrate the efficacy and safety of intraperitoneally applied phospholipids with regard to adhesion prevention and wound healing, respectively. The assumption is that phospholipids rapidly adhere to the peritoneal surface and to the mesothelial lesions. This study was designed to evaluate the influence of early drainage of the administered fluid volume on the control of adhesion formation. Forty chinchilla rabbits underwent median laparotomy and standardized abrasion of circumscript areas of the ventral abdominal wall, the cecum, and the ileum. The animals randomly received either 75 mg/kg body weight of phospholipids in a volume of 5.0 mL/kg body weight (n = 20) or the same volume of Ringer's lactate solution (n = 20) prior to closing the laparotomy wounds. In 50% of the rabbits with either medication, 80% of the volume was recovered after 30 min before final closure of the abdominal wall ("drainage"). In the remaining animals the intraabdominal fluid load was not evacuated ("no drainage"). At day 10 after surgery all rabbits were sacrificed for evaluation of adhesion areas by computer-aided planimetry and histopathologic examination. The mean areas of adhesion in both Ringer's lactate groups were significantly larger than in the comparable phospholipid groups (p < .05). In the Ringer's lactate groups, adhesions averaged 341.7 (318.6) mm2 without and 263.3 (275.5) mm2 with drainage. In the phospholipid groups the respective mean areas reached only 24.6 (36.7) mm2 without drainage and 27.0 (49.7) mm2 following evacuation of the fluid 30 min after administration (median, mean in parentheses). These results prove the efficacy of phospholipids after a limited contact period of 30 min. The frequent use of drains in abdominal surgery will not impair the beneficial effect of phospholipids on prevention of adhesions. PMID- 11931491 TI - Proinflammatory cytokines are not released in the circulation following acute pulmonary thromboembolism in pigs. AB - Histological examination of acute lung injury associated with sepsis often revealed thromboembolic lesions in the pulmonary microcirculation. Several inflammatory mediators such as platelet activating factor, thromboxane, and endothelins have also been implicated in the pathogenesis of acute pulmonary thromboembolism (APTE). In the present study we examined the roles of three proinflammatory cytokines, tumor necrosis factor-alpha (TNF-alpha), interleukin 1beta (IL-1beta), and IL-8, in the early phase of APTE. APTE was induced in 13 anesthetized piglets (22+/-4 kg) by injecting thrombin-induced blood clots directly into the left lower lobar pulmonary artery. Five animals that received only warm sterile saline served as controls. Arterial plasma samples were collected regularly over 8 h so that cytokine levels could be measured later by enzyme-linked immunosorbent assay (ELISA). Administration of clots doubled the mean pulmonary arterial pressure (from 13+/-5 to 26+/-7 mm Hg) and caused significant decrease in arterial oxygen tension (PaO2 from 390+/-85 to 256+/-89 mm Hg while the FiO2 was maintained at 1.0). Mean arterial blood pressure and cardiac output remained comparable throughout the experiments after initial fluid resuscitation. Plasma levels of TNF-alpha, IL-1beta, and IL-8 were not significantly increased in the APTE group when compared with their baseline values or the control group. Our results thus show that APTE is associated with pulmonary hypertension and deterioration of gas exchange but not with the systemic release of TNF-alpha, IL-1beta, or IL-8. We conclude that these cytokines have minimal impact on the systemic circulation during APTE. PMID- 11931492 TI - Visionary mind of a surgical scientist: Marie Joseph Auguste Carrel. PMID- 11931493 TI - Adhesion formation: intraperitoneal catheters in surgical practice. AB - To compare the adhesion formation, in a rat model, of commercially available materials used as intraperitoneal catheters in clinical practice, pieces of polyurethane, Teflon, silicone, and polyvinyl chloride (PVC) were placed intraperitoneally in 36 female Wistar rats randomly assigned into four groups. Adhesion development was quantitated according to Nair's classification system (NCS). According to NCS, polyurethane showed the least adhesion formation, while the highest scores were found in the PVC group. Comparing the thickness and the firmness of adhesions in the four groups, significant differences were found, with the polyurethane group developing none or filmy and mild adhesions. In contrast, almost all the rats in the Teflon group developed thick and dense adhesions. Thus, polyurethane caused the least adhesion formation among the four materials evaluated, and can be used more extensively for the construction of catheters for prolonged intraperitoneal use. PMID- 11931494 TI - Surgical experience with retroperitoneal heterotopic heart transplantation in the large white domestic swine. AB - Accelerated coronary atherosclerosis following heart transplantation is the main limiting factor for long-term survival, aside from graft failure and complications due to immunosuppression. Graft coronary vasculopathy is due to chronic rejection of the vascular wall leading to intimal hyperplasia in coronary arteries. Numerous heterotopic heart transplantation models have been used in different species to study the immunology and pathophysiology following graft implantation. This study reports our experience with the retroperitoneal heterotopic heart transplantation in Large White domestic swine using immunological typing. This approach mimics the kinetics of slow low-grade rejection in clinical human heart transplantation. One hundred and fifty-four retroperitoneal (n = 154) heterotopic heart transplantations were performed using Large White swine sampled for the major histocompatibility class (MHC) class I antigen and blood type using the microlymphotoxicity technique. Acute rejection studies were performed by intentional mismatch of the swine lymphocyte alloantigen (SLA) and chronic rejection studies were done in allografts implanted in donor-recipients matched for blood type and class I antigen to assess the effects of rejection per se, hypercholesterolemia, intracoronary L-NAME infusion, and endothelial denudation on the development of graft coronary vasculopathy. Assessment of in vitro coronary arterial vascular reactivity in standard organ chamber experiments comprised the core of vascular biology studies in this large animal model. Eighty (52%) transplanted recipients survived until the elective date of sacrifice at 60 days, 14 (9.1%) died during the surgery, 21 (13.6%) died <24 h after the transplant, and 8 (5.2%) died of late deaths. The retroperitoneal heterotopic heart transplantation model with blood typing and determination of the SLA class I antigen is a useful model for the study of immunological and vascular events due to graft rejection after heart transplantation. PMID- 11931495 TI - Response to nitric oxide during adult cardiac surgery. AB - Pulmonary hypertension is associated with significant morbidity and mortality in adult cardiac surgery patients. Inhaled nitric oxide is known to be a selective pulmonary vasodilator in this setting. However, it is not known which cardiac surgery patients benefit most from nitric oxide therapy. This study sought to prospectively determine whether a patient's baseline pulmonary vascular resistance could be used to predict responsiveness to inhaled nitric oxide therapy. Subjects were 30 consecutive cardiac surgery patients with pulmonary hypertension immediately prior to induction of anesthesia. There were 2 study groups: Group 1 (n = 15) had an initial pulmonary vascular resistance between 125 and 300 dyn-s/cm5, while group 2 (n = 15) had an initial pulmonary vascular resistance of greater than 300 dyn-s/cm5. Both groups were empirically treated with inhaled nitric oxide (30 ppm) upon separation from bypass. The conduct of anesthesia, surgery, and cardiopulmonary bypass were controlled. A therapeutic algorithm dictated the use of vasoactive substances for all patients. Heart rate, mean arterial pressure, pulmonary vascular resistance, peripheral vascular resistance, cardiac index, and right ventricular ejection fraction were monitored throughout the operative experience. Patients with a higher initial pulmonary vascular resistance had a significantly greater percent reduction in pulmonary vascular resistance after the initiation of nitric oxide therapy. This study suggests that pulmonary vascular resistance is more dramatically affected by inhaled nitric oxide in cardiac surgery patients with a greater degree of pulmonary hypertension. PMID- 11931496 TI - Pesticide mortality in the United States 1979-1998. AB - Pesticide mortality in the US is usualy reported on a case-by-case basis. The Vital Statistics of the US and the publications of the American Association of Poison Control Centers publish yeary statistics on pesticide mortality. This review evaluates the incidence of pesticide mortality in regard to intent, geography, sex, race, age and trends from theyears 1979-1998. In this fashion it appeared easier to identify likely areas of exposures and to develop steps to reduce mortality. Pesticide mortality has decreased substantially over the last 20 y. Intentional poisonings, primarily suicides, represent the greatest fraction and are decreasing more slowy than accidental poisonings. Mortality is thus following intentional exposure rather than accidental exposure. Intentional exposures may occur away from sites where pesticide use is expected. PMID- 11931497 TI - A prospective study of multiple supratherapeutic acetaminophen doses in febrile children. AB - Repeated doses of acetaminophen given for therapeutic reasons have been reported to cause hepatotoxicity in adults and children. We studied the effect of repeated acetaminophen (APAP) overdoses administered for therapeutic purposes in a prospective cohort of children. Forty-four children, aged 2 mo to 10 y were referred with a fever of >38.5 C for more than 48 h, and received >60 mg APAP/kg/d. In each patient AST, ALT and APAP blood levels were measured. The mean total daily dose of APAP was 92+/-26 (63-171) mg/kg. There was a weak, but significant, negative correlation between age and daily dose of APAP where younger children received higher doses of APAP. In 4 children (9.1%) an elevation of AST and ALT was found. Three of the 4 patients with elevated liver enzymes had received >90 mg APAP/kg/day; APAP blood levels ranged from 0 to 23 mg/mL. No correlation was found between the time since last APAP dose and the serum drug level. Ill children receiving repeated supratherapeutic doses of acetaminophen may show abnormalities in liver function. However, severe liver injury was rare. PMID- 11931498 TI - Missing the point! AB - A medication order for warfarin sodium was sent by facsimile transmission with a decimal point and trailing zero. The order was incorrectly transcribed as a ten fold overdose and would almost certainy have been fatal if administered as transcribed. Other hazardously written medication orders are presented with guidelines for safer ordercomposition. PMID- 11931499 TI - Incidence of intentional poisoning of dogs in the Abruzzo region of Italy. AB - Toxicological assays done between 1997 and 2000 on 105 presumed baits of the baits and poisonings in 408 dogs are reported. Of the baits, 54.4% were positive for organophosphates and 27.5% for strychnine. In 31.1% of the dog cases, analysis confirmed the presence of toxic concentrations of organophosphate pesticides (43.3%) and strychnine (44.1%). PMID- 11931500 TI - Aflatoxin in dog and horse feeds in turkey. AB - Aflatoxin levels were determined by ELISA in 18 dog and 20 horse feed samples, collected from different firms from June 2000 to June 2001 in Turkey. The minimum and maximum levels of total aflatoxin in the dog and horse feeds were <1.75-20 microg/kg and <1.75-14 microg/kg, respectively; 3/18 dog feed samples (16.7%) and 2/20 horse feed samples (10%) exceeded the Turkish tolerance limit of 10 microg/kg in food or feed. PMID- 11931501 TI - Common causes of poisoning in dogs and cats in a Brazilian veterinary teaching hospital from 1998 to 2000. AB - A retrospective study at the University Veterinary Teaching Hospital of Sao Paulo city, Brazil, describes the pattern of poisoning that occurred from 1998 to 2000. During this period 5,136 animals were sent to the emergency sector, and 250 of these cases were reported as poisonings: 203 dogs (81.2%) and 47 cats (18.8%). The common causes of poisoning in dogs were 28.9% therapeutic products (86.4% non steroidal anti-inflammatory drugs, 6.8% other drugs, 3.4% antibiotics, 3.4% tranquilizing agents), 15.8% rodenticides, 13.9% pesticides for farm use (39.3% organophosphorous and 35.7% carbamate insecticides, 25.0% amitraz), 11.8% unknown agents, 8.4% plants, 6.8% industrial products, and 5.0% pesticides for domestic use. The common causes of poisoning in cats were 29.9% therapeutic products (50.0% non-steroidal anti-inflammatory drugs, 42.8% others, 7.2% antibiotics), 27.6% pesticides for farm use (46.1% carbamate insecticides, 38.5% organophosphorous insecticides, 15.4% others), 14.9% pesticides for domestic use, 12.8% unknown agents, 10.6% rodenticides, and 4.2% industrial products. These data show it is essential to create an awareness of the toxicity of these agents to reduce poisoning incidence. PMID- 11931502 TI - It's deja vu--all over again! PMID- 11931503 TI - Atypical presentation of risperidone toxicity. PMID- 11931504 TI - Pediatric environmental health specialty units (PEHSUs) in 2002! PMID- 11931505 TI - Effects of dietary sulfur concentrations on the incidence and pathology of polioencephalomalicia in weaned beef calves. AB - Fourteen heifer calves weighing 174.5+/-17.7 kg were used to evaluate the effects of 3 levels of dietary sulfur. Sodium sulfate added to basal diet made treatments designated moderate (3860 ppm sulfur), moderatey high (5540 ppm sulfur) and high (7010 ppm sulfur). Clinical polioencephalomalacia occurred in all calves assigned to the moderately high and high treatments. The calves did not acclimate to the dietary sulfur as polioencephalomalacia occurred in 4 animals on d 35 and in 1 calf on d 37. Microscopic lesions confirmed polioencephalomalacia in the calves on moderately high and high diets. Microscopic lesions also were present in 4 moderate diet calves although clinical signs were not seen. High dietary sulfur did not limit feed intake. Diets containing sulfur levels >4000 ppm sulfur produced polioencephalomalacia in 10 calves and sub-clinical brain lesions occurred in 4 calves consuming <4000 ppm sulfur. PMID- 11931506 TI - Effects of deltamethrin on lipid peroxidation in mice. AB - Effects of deltamethrin on lipid peroxidation were investigated for subacute, subchronic and chronic periods. Deltamethrin was given to Swiss Albino mice at 1.5, 2.5, or 7.5 mg/kg. Lipid peroxidation was evaluated by determining malondialdehyde (MDA) levels in plasma, and determining glutathione peroxidase (GSH-Px), superoxide dismutase (Cu-Zn SOD) and catalase activities in erythrocytes on days 15, 45, and 60. MDA levels increased in deltamethrin dosed groups, especially for the subchronic and chronic periods. GSH-Px, Cu-Zn SOD and catalase activities in erythrocytes were decreased at high doses of deltamethrin, especially at the high dose (7.5 mg/kg) deltamethrin group. PMID- 11931507 TI - Neuropsychological studies on lead battery workers. AB - This study assessed the psychological status of 60 lead-acid battery workers occupationally exposed to low level lead and 30 referents in Lucknow. Digit spans and symbol, Bourdon-wiersma vigilance and Raven's Progressive matrices tests were administered. Significant impairments in concentration, attention, auditory and visual memory, psycho-motor speed, perceptual accuracy, and visual reasoning were observed in the workers compared to the referents. The deficits were not related to exposure duration since the magnitude of the impairments observed in workers with 1-y duration was the same as in those with more (up to 30 years) work duration. The blood lead of the battery workers was significantly elevated. There were functional deficits of the central nervous system in the lead-acid battery workers. PMID- 11931508 TI - Depletion and bioavailability of imidocarb residues in sheep and goat tissues. AB - The residual depletion of a commercial product containing imidocarb dipropionate in sheep and goat tissues was investigated. Additionally, the oral bioavailability of residues was determined in rats to evaluate the extent to which tissue imidocarb residues could be reabsorbed by consumers. Ten ewes and 5 goats were administered im with a commercial formulation containing imidocarb dipropionate (Carbesia cavalli, Shering-Ploug 121.15 mg/ml) at the single dose of 3 mg/kg bw corresponding to 2.1 mg/kg bw imidocarb base. Two sheep and 1 goat were slaughtered 15, 30, 60, 90 or 120 d after dosing and samples of muscle, injection site muscle, liver, omental and subcutaneous fat, and kidneys were collected. Samples of cerebral hemisphere, cerebellum, olfactory bulb, pineal and pituitaryglands were dissected. For the residue bioavailability study 7 groups of3 Wistar rats each, were dosed by gavage with imidocarb dipropionate standard in water (group 2, 3 and 4) or with imidocarb as a liver residue collected from prior dosed animals (group 5, 6 and 7) at 8.4. 16.8 or 33.6 microg/kg of imidocarb base respectively, for 5 d. Group I was control. All animals were sacrificed the day after the last drug administration and livers were collected. The highest drug levels in sheep and goats occurred in liver and kidney, suggesting that these tissues are targets for residues; muscle had negligible importance as storage tissue. Goats had a lower storage capability than sheep. The residue profile in sheep liver and omental fat showed a 30-d storage period to reach maximum concentrations, and suggested that imidocarb is redistributed. The high and long-lasting concentrations in brain showed its capacity to cross the blood-brain barrier and caused concern for potential neurotoxic effects. Detectable concentrations of imidocarb were not found in rat liver. PMID- 11931509 TI - Suppressive effect of zearalenone, an estrogenic mycotoxin, on bovine neutrophil chemiluminescence. AB - The effects of zearalenone (ZEA), an estrogenic mycotoxin produced by Fusarium fungi, on bovine neutrophils were investigated in vitro using chemiluninescence, a bactericidal parameter. ZEA suppressed luminol-dependent, phorbol 12-myristate 13-acetate (PMA)-elicited chemiluminescence in a dose-dependent manner at concentrations of 10(-4) M and 10(-5) M. No significant suppression was observed at concentrations lower than 10(-6) M. The possible mode ofaction of 10(-4) M ZEA on the cell activity was investigated with special reference to intracellular Ca2+ ([Ca2+]i) release and estrogen receptors. The 10(-4) M ZEA treatment significantly impaired [Ca2+]i release. When pretreated with a low dose (10(-6) M) of PMA, the cells resisted the ZEA-induced chemiluminescence suppression. However, pretreatment of the cells with the estrogen receptor blockers Tamoxifen and ICl 182,780 (both at 10(-6) M) did not annul the suppressive ZEA action. Considering that PMA is an activator of protein kinase C (PKC), a signal transducing enzyme, and in association with a rise in [Ca2+]i causes cytosolic PKC to shift to the plasma membrane where the activated PKC triggers a varied array of cellular responses, the pharmacological dose of ZEA might have suppressed chemiluminescence by hindering the release of [Ca2+]i and the PKC shift. The results of pretreatment with estrogen receptor blockers, however, did not support the suggestion that the ZEA treatment affected the cells via estrogen receptor pathways. PMID- 11931510 TI - Hepatotoxicity caused by Breynia officinalis. AB - Breynia officinalis has the Chinese proprietary name, Chi R Yun, which means dizziness or vertigo for 7 d. In daily practice, it has been used to treat venereal diseases, contusion, heart failure, growth retardation and conjunctivitis in combination with other traditional Chinese medicines. Two hospital-based cases of Breynia officinalis poisoning have been reported to the Poison Control Center. Case 1 was a 43-y-old female who consumed a mixture of 1500 g lower stem and root of Ji Mu Ju in boiled water in a suicide attempt. Her AST reached 264 and ALT reached 2443. Case 2 was a 51-y-old female who consumed 20 pieces of lower stem and root of Ji Mu Ju stewed with meat and 100 ml of wine to treat chronic contact dermatitis. Her AST reached 3815 and ALT reached 6625. In both cases Breynia officinalis was identified as the cause of poisoning. Poisoning in humans involves the neurologic, gastrointestinal, hepatic, urinary and respiratory systems. Hepatotoxic effects have been reported for some Chinese herbal medicines, but not Breynia officinalis: Breynia officinalis poisoning causes hepatocellular liver injury rather than cholestatic liver injury. PMID- 11931511 TI - Chlorhexidine gluconate ingestion resulting in fatal respiratory distress syndrome. AB - As ingestion of chlorhexidine gluconate (CHG) usually causes relatively mild symptoms, this chemical has been considered safe. An 80-y-old woman with dementia accidentally ingested approximately 200 ml of Maskin (5% CHG) in a nursing home and then presumably aspirated gastric contents. She was intubated for airway protection in the nearest hospital and referred to our critical care unit because of hypotension and rapid deterioration of consciousness. Despite intensive treatment, the patient died of acute respiratory distress syndrome (ARDS) 12 h after ingestion. The serum concentration of CHG was markedly high, although CHG reportedly has poor enteral absorption. We suspect the CHG was absorbed through the pulmonary alveoli following aspiration, not from the gastrointestinal tract. CHG has the potential for fatal ARDS when aspiration occurs following ingestion. PMID- 11931512 TI - High-dose montelukast exposures in a 3-year-old and a 5-year-old child. AB - The leukotriene receptor antagonists (LTRAs) is a relatively new class of asthma medication with a lack of toxicity for unintentional poisoningsituations. This makes it difficult to determine which exposures require aggressive decontamination or simple monitoring in the home setting: prompting the question, "What LTRA dose is likely to produce significant toxicity?" We report a case of an unintentional poisoning with 80 mg montelukast in a 3-y-o asthmatic child that was managed in the home with observation alone and a second case of untentional 135 mg montelukast poisoning in a 5-y-o asthmatic child managed in an emergency department. In both cases, symptoms were not observed. These cases and available literature suggest that doses < 4.5 mg/kg in children result in minimal toxicity. PMID- 11931513 TI - Intoxication with sodium monofluoroacetate (compound 1080). AB - The highly toxic sodium monofluoroacetate (SMFA) was banned as a rodenticide in the U.S. in 1972. We report the first case of intentional ingestion in this country in over 15y. A 47-y-old male was brought to the emergency room status post tonic clonic seizure. At 34 h post ingestion, he responded ony to noxious stimuli and at 48 h, he was unresponsive to painful stimuli, was intubated and placed on a ventilator. Over the following 3 d, he was became minimally responsive to external stimuli with bouts of agitation and hypertension. Two days later he was discharged with no evidence of neurologic sequelae. We report this patient to increase awareness of SMFA toxicity, and its ability to cause anion gap metabolic acidosis. PMID- 11931514 TI - Amanita mushroom poisoning: efficacy of aggressive treatment of two dogs. AB - Amatoxins, the primary toxins found in mushrooms of the genus Amanita, are very toxic to dogs. Acute fulminant liver failure and death can occur within a few days of ingestion. By their curious nature, dogs, especially young dogs, are prone to ingest mushrooms. Early identification of suspect mushrooms, and prompt emergency measures aimed at decreasing absorption of the toxins can improve the chance of survival. Knowing the major clinical syndromes associated with Amanita mushroom toxicosis can help direct the treatment and supportive care of affected animals and improve survival rates. We describe 2 cases in dogs with confirmed ingestion of Amanita phalloides and Amanita ocreata resulting in fulminant liver failure. Death occurred in 1 dog despite aggressive treatment measures including hemoperfusion, while aggressive measures resulted in a favorable outcome in the other dog. PMID- 11931515 TI - Anticholinesterase-induced hypotension treated with pulmonary artery catheterization-guided vasopressors. AB - We report a case of severe hypotension caused by anticholinesterase poisoning that was successfully treated with high-dose vasopressor therapy guided by pulmonary artery catheterization. PMID- 11931516 TI - On the universality and cultural specificity of emotion recognition: a meta analysis. AB - A meta-analysis examined emotion recognition within and across cultures. Emotions were universally recognized at better-than-chance levels. Accuracy was higher when emotions were both expressed and recognized by members of the same national, ethnic, or regional group, suggesting an in-group advantage. This advantage was smaller for cultural groups with greater exposure to one another, measured in terms of living in the same nation, physical proximity, and telephone communication. Majority group members were poorer at judging minority group members than the reverse. Cross-cultural accuracy was lower in studies that used a balanced research design, and higher in studies that used imitation rather than posed or spontaneous emotional expressions. Attributes of study design appeared not to moderate the size of the in-group advantage. PMID- 11931517 TI - Methodological requirements to test a possible in-group advantage in judging emotions across cultures: comment on Elfenbein and Ambady (2002) and evidence. AB - H. A. Elfenbein and N. Ambady's (2002) conclusions concerning a possible in-group advantage in judging emotions across cultures are unwarranted. The author discusses 2 methodological requirements for studies to test adequately the in group advantage hypothesis and an additional requirement in reviewing multiple judgment studies and examining variance in judgment effects across those studies. The few studies that Elfenbein and Ambady reported that support the in-group advantage hypothesis need to be examined for whether they meet the criteria discussed; if they do not, their data cannot be used to support any contention of cultural differences in judgments, let alone the in-group advantage hypothesis. Furthermore, the role of signal clarity needs to be explored in possibly moderating effects across studies; however, this was not done. PMID- 11931518 TI - Is there an in-group advantage in emotion recognition? AB - H. A. Elfenbein and N. Ambady (2002) examined the evidence for an in-group advantage in emotion recognition, whereby recognition is generally more accurate for perceivers from the same cultural group as emotional expressors. D. Matsumoto's (2002) comment centered on 3 asserted methodological requirements. This response addresses the lack of consensus conceming these "requirements" and demonstrates that none alter the presence of the in-group advantage. His analyses had a serious flaw and, once corrected, replicated the original findings. Furthermore, he described results from his empirical work not meeting a literal interpretation of his own requirements. Overall, where Matsumoto considers subtle cross-cultural differences in emotional expression a methodological artifact in judgment studies, the present authors find a core phenomenon worthy of attention. PMID- 11931519 TI - From infant to child: the dynamics of cognitive change in the second year of life. AB - The authors review several key areas of early cognitive development in which an abrupt shift in ability at the end of the second year of life has been traditionally assumed. These areas include deferred imitation, self-recognition, language, and categorization. Contrary to much conventional theorizing, the evidence shows robust continuities in all domains of early cognitive development. Where there is evidence of a reorganization of behavior that makes a new level of performance possible, dynamic-systems analyses indicate that even these may be driven by underlying processes that are continuous. Although there remain significant definitional and methodological issues to be resolved, the outcome of this review augers well for newer models in which cognitive development is viewed as a continuous, dynamic process. PMID- 11931520 TI - Multiple cues for quantification in infancy: is number one of them? AB - A review and synthesis of the literature on quantification in infancy and early childhood is provided. In most current conceptualizations, early quantification is assumed to be number based. However, the extant literature provides no clear cut evidence that infants use number to perform quantitative tasks. Instead, new research suggests that quantification is initially based on nonnumerical cues, such as area and contour length, whether or not a task involves discrete items. The authors discuss the implications of these findings with respect to early quantification and its relation to later numerical development. PMID- 11931521 TI - Socioeconomic differences in children's health: how and why do these relationships change with age? AB - The effects of socioeconomic status (SES) on health are well documented in adulthood, but far less is known about its effects in childhood. The authors reviewed the literature and found support for a childhood SES effect, whereby each decrease in SES was associated with an increased health risk. The authors explored how this relationship changed as children underwent normal developmental changes and proposed 3 models to describe the temporal patterns. The authors found that a model's capacity to explain SES-health relationships varied across health outcomes. Childhood injury showed stronger relationships with SES at younger ages, whereas smoking showed stronger relationships with SES in adolescence. Finally, the authors proposed a developmental approach to exploring mechanisms that link SES and child health. PMID- 11931522 TI - Risky families: family social environments and the mental and physical health of offspring. AB - Risky families are characterized by conflict and aggression and by relationships that are cold, unsupportive, and neglectful. These family characteristics create vulnerabilities and/or interact with genetically based vulnerabilities in offspring that produce disruptions in psychosocial functioning (specifically emotion processing and social competence), disruptions in stress-responsive biological regulatory systems, including sympathetic-adrenomedullary and hypothalamic-pituitary-adrenocortical functioning, and poor health behaviors, especially substance abuse. This integrated biobehavioral profile leads to consequent accumulating risk for mental health disorders, major chronic diseases, and early mortality. We conclude that childhood family environments represent vital links for understanding mental and physical health across the life span. PMID- 11931523 TI - Leptin and obesity in mother-child pairs. AB - Defects in the leptin gene or the leptin receptor may be a genetic cause of obesity, but little is known about the familial associations of leptin and obesity. This study compared plasma leptin and measures of obesity in a sample of 248 subjects (124 mother-offspring pairs); 34% were African American and 66% were white. Youth were aged 12 to 16 years. Plasma leptin and body mass index (BMI) were higher in mothers than in their offspring and, among the offspring, higher in girls than boys, even after correcting for BMI or body fat. Racial differences in leptin were present in both mothers and youth when adjusting for percentage body fat but disappeared when adjusting for BMI. In univariate analyses of the associations between mothers and offspring, BMI was associated with leptin in all groups but was most strongly associated in white pairs and in mother-son pairs. In multiple regression analyses, when adjusting for BMI, significant predictors of leptin levelfor the boys and girls together were gender, BMI, and pubertal status of the offspring; in girls only BMI was significant (R2 = 0.72), and in boys the significant predictors were their BMI (R2 = 0.66) followed by their pubertal status (R2 = 0.06) and the leptin level of their mothers (R2 = 0.02). When adjusting for body fat, the predictors were the offspring's percentage bodyfat (R2 = 0. 67) and mother's leptin (R2 = 0.03), with similar results in gender-specific analyses. The authors conclude that leptin levels of youth are most closely associated with their degree of obesity or body fat; mother's leptin and, for boys only, pubertal status also play a small role. Although the small association between maternal leptin on leptin in their offspring could be due to either heredity or shared environment, the results of this study suggest that individual obesity and environmental factors are important predictors of leptin levels in children. PMID- 11931524 TI - Computer-assisted continuous ST-segment analysis for clinical research: methodological issues. AB - Continuous ST-segment monitoring has been used to detect acute myocardial ischemia, determine the success of the reperfusion therapy, and predict outcomes in both research and a variety of clinical settings. However, analyzing the abundant electrocardiography (ECG) data recorded using continuous multilead ST segment monitoring techniques is time consuming and requires expertise. Experienced data interpreters in dedicated ECG core laboratories handle many continuous ECG data records from large clinical trials. Little information on measurement issues for computer-assisted ST-segment analysis is available for individual investigators. Unsupervised or inexperienced computer analysis of ST segment deviations can, under certain circumstances, yield invalid or unreliable summary indices. The goal of this article is to discuss basic ST-segment measurement principles in evaluating acute myocardial ischemia and methodological issues surrounding the use of computer-assisted ST-segment analysis for continuous ECG data. Variables affecting ST-segment measurements will be examined. Sources and examples of variability for these potential errors will be identified. PMID- 11931525 TI - Instillation of 3% hydrogen peroxide or distilled vinegar in urethral catheter drainage bag to decrease catheter-associated bacteriuria. AB - Patients with indwelling catheters acquire urinary tractinfections ata rate of 5% perday. After 30 days of catheterization, there is a 78% to 95% incidence of bacteriuria, despite the use of meticulous catheter care. Research studies on reducing the development of bacteriuria have focused on the propagation of bacteria within the catheter drainage bag. This research proposal sought to determine the effects of instillation of 3% hydrogen peroxide versus distilled vinegar in urethral catheter drainage bags to decrease bacteriuria in 20 long term catheterized patients. Baseline urine cultures for control and test mediums showed no significant difference in rates of bacteriuria. However, urine cultures obtained at the 48-hour interval showed significant reduction in bacteriuria in urinary bags irrigated with vinegar The implications this study has for nursing practice include a needforfurther research using a larger number of subjects and a need for improvements in the design of the catheter drainage system. PMID- 11931526 TI - Effects of inactivity on glycolytic capacity of single skeletal muscle fibers in adult and aged rats. AB - The purpose of this study was to determine the effects of inactivity on lactate dehydrogenase (LDH) enzyme activity (expressed in nmol/g dry weight x hour) in single skeletal musclefibers from the soleus muscle in adult and aged rats. Fourteen 12-month-old andfifteen 30-month-old Fisher 344 Brown Norway F1 Hybrid rats were randomly assigned to control, 1 week of hindlimb unweighting (HU1), or 2 weeks of hindlimb unweighting (HU2). With age, a significant decrease in LDH enzyme activity occurred in type I skeletal muscle fibers (29.5%, P < 0.05). Following HU2, individual type I skeletal muscle fibers from the 12-month-old animals showed a 33.3% increase in LDH activity. In contrast, individual type I fibers from the aged animals showed a 50.0% increase after HU1. In conclusion, the baseline levels of LDH activity were significantly less in aged versus adult rats. The timing of the skeletal muscle adaptation to inactivity was different between young and old animals, such that the older animals responded to inactivity before the younger animals. These biochemical changes may have an impact on the fatigability of the muscle following inactivity. Thefindings indicate that treatment during bed rest for the older adult may be different than that for the younger adult. PMID- 11931527 TI - Gastrointestinal cancer of the microsatellite mutator phenotype pathway. PMID- 11931529 TI - Transient increase of rat gastric amylin in the neonatal period and in experimental ulcers. AB - BACKGROUND: Amylin (islet amyloid polypeptide; IAPP)-containing cells are present in the gastric mucosa and duodenum, but little is known about their regulation. In the present study, we investigated rat gastric mucosa in the neonatal period and in an experimental model of gastric ulcer. METHODS: Gastric mucosa was fixed in Bouin's solution and cut into 5-microm sections, which were examined for amylin-containing cells by light microscopic morphometry, using specific antisera and the peroxidase-antiperoxidase (PAP) method. RESULTS: In the neonatal period (days 1, 7, 18, 28, and 45; five animals at each time point), small numbers of amylin-positive cells were consistently found in the basal cell layer of the mucosa, from birth throughout the study. A marked, transient increase in amylin positive cells was noted in all animals examined on day 18, the day after the introduction of pellet feed. In the second set of experiments, acetic acid induced gastric ulcers were examined. Necroticulcer lesions were observed on day 4 after the acetic acid injection, associated with increases in amylin-positive cells in the vicinity of the ulcer margins. Amylin positivity peaked on day 6 and had returned to the control level by day 28. CONCLUSIONS: The results indicate that gastric amylin increases in response to mucosal stimuli and may suggest a role of amylin in mucosal growth and repair. PMID- 11931528 TI - Irritant-induced cyclooxygenase-2 is involved in the defense mechanism of the gastric mucosa in mice. AB - BACKGROUND: Endogenous and exogenous prostaglandins (PGs) have been shown to contribute to reducing the gastric injury caused by irritants given subsequently. The aim of this study was to clarify whether cyclooxygenase-2 (COX-2) protein induced by pretreatment was involved in the prevention of subsequent ethanol caused gastric injury in mice. METHODS: Mice were pretreated with acidified ethanol or saline and then COX-2 protein expression in the stomach was immunohistochemically determined every 8h. Mice were administered 95% ethanol 24h after the acidified ethanol pretreatment, and gastric mucosal damage was evaluated macroscopically and histologically. The effects of NS-398 or indomethacin on the 95% ethanol-caused damage were also examined. RESULTS: Acidified ethanol pretreatment induced COX-2 protein expression in lamina propria macrophages of the gastric mucosa, with a peak level 24h after the pretreatment. The 95% ethanol treatment caused gastric mucosal damage. The degree of the damage was not different between mice pretreated with acidified ethanol and those pretreated with saline. However, NS-398 aggravated the ethanol-caused damage only in mice pretreated with acidified ethanol, while indomethacin aggravated the damage, evaluated histologically, irrespective of the pretreatment. CONCLUSIONS: Pretreatment-induced COX-2, in addition to COX-1, seemed to be involved in the defense mechanism through minimizing the damage caused by a subsequent irritant. PMID- 11931530 TI - The effects of rabeprazole on parietal cells and enterochromaffin-like cells in rats: a comparison with omeprazole. AB - BACKGROUND: We investigated the effects of rabeprazole compared with those of omeprazole on enterochromaffin-like cells and parietal cells in rats. METHODS: Rabeprazole or omeprazole was administered for 7 days by intraperitoneal injection (100 mg/kg or 20mg/kg once a day) and the serum gastrin concentration, the antral density of G cells and D cells, fundic histamine content, fundic H+, K+-ATPase mRNA level, and parietal cell morphology were determined. RESULTS: Both rabeprazole and omeprazole inhibited gastric acid secretion and increased the intragastric pH to over 6.5, as well as causing a marked increase in the serum gastrin concentration. The serum gastrin level was lower with rabeprazole treatment than with omeprazole treatment at both doses. Also, the antral G-cell density was higher with omeprazole than with rabeprazole, while the increase in both the histamine content and the H+, K-ATPase mRNA level in the fundic mucosa was higher with omeprazole treatment at both doses, with the difference being significant at 100 mg/kg. Ultrastructural examination indicated that the stimulation of parietal cells by omeprazole was stronger than that by rabeprazole. CONCLUSIONS: Rabeprazole treatment does not drive enterochromaffin like cells and parietal cells as strongly as omeprazole treatment despite its potent acid suppressive effect, suggesting that it represents a new generation of proton pump inhibitors. PMID- 11931531 TI - Effects of cyclooxygenase-2 inhibitor NS-398 on APC and c-myc expression in rat colon carcinogenesis induced by azoxymethane. AB - BACKGROUND: Nonsteroidal anti-inflammatory drugs (NSAIDs) have been reported to protect against the development of colon cancer; however, the mechanism(s) by which NSAIDs exert their effects is still unclear. The aim of this study was to examine the effects of NSAIDs on the expression of the tumor suppressor APC gene and the c-myc oncogene in the colons of rats treated with a colon-specific carcinogen, azoxymethane (AOM). METHODS: Gene expression levels were estimated by a reverse transcription (RT)-competitive polymerase chain reaction (PCR) method. RESULTS: The group of rats simultaneously administered AOM and NS-398, a cyclooxygenase (COX)-2 inhibitor, showed a significant reduction in the number of preneoplastic lesions of colon cancer compared with that in the group of rats treated with AOM alone. Furthermore, the APC expression level in the group of rats treated with both AOM and NS-398 was significantly greater than that in the group of rats treated with AOM alone; this result for APC gene expression was reconfirmed by the immunohistochemical staining of APC protein. In addition, c myc mRNA expression was clearly decreased in the group of rats treated with both AOM and NS-398 compared with the level in the group of rats treated with AOM alone. CONCLUSIONS: Our data thus indicate that NS-398 causes an increase in APC expression and a decrease in c-myc expression. PMID- 11931532 TI - One-week losartan administration increases sodium excretion in cirrhotic patients with and without ascites. AB - BACKGROUND: Losartan, a highly selective angiotensin II type 1 receptor antagonist, has been reported to have a significant portal hypotensive effect in cirrhotic patients. A recent study also showed that losartan exerted a dramatic natriuretic effect in preascitic cirrhosis. The influence of losartan on renal hemodynamics and sodium homeostasis in cirrhotic patients with ascites is unclear. This study was undertaken to evaluate the renal effects of 1-week losartan treatment in cirrhotic patients with and without ascites. METHODS: All 12 patients in the study received a daily oral dose of 25 mg losartan for 7 consecutive days. Effective renal plasma flow, urine volume, creatinine clearance, 24h urine sodium excretion and fractional excretion of sodium, blood urea nitrogen, and serum creatinine were measured before and after treatment. RESULTS: In cirrhotic patients without ascites, creatinine clearance, 24-h urinary sodium excretion, and fractional excretion of sodium were significantly increased after losartan administration. Effective renal plasma flow and serum creatinine showed almost no change after treatment. In cirrhotic patients with ascites, creatinine clearance, 24-h urinary sodium excretion, fractional excretion of sodium, and effective renal plasma flow were significantly increased after losartan administration. In addition, the magnitudes of the increases in the fractional excretion of sodium and in the 24-h urinary sodium excretion were greater in cirrhotic patients with ascites than in those without ascites. CONCLUSIONS: One-week treatment with losartan increases sodium excretion in association with an improvement of renal function in cirrhotic patients with and without ascites. The natriuretic effect was more profound in cirrhotic patients with ascites than in those without ascites. PMID- 11931533 TI - Diagnosis of pT2 gallbladder cancer by serial examinations with endoscopic ultrasound and angiography. AB - BACKGROUND: The prognosis of pT2 gallbladder cancer correlates with whether appropriate surgery for the spread of cancer has been performed. Therefore, accurate preoperative T staging is especially important. We carried out this study to evaluate the usefulness of serial examinations by endoscopic ultrasound (EUS) and angiography for the T staging of pT2 gallbladder cancer. METHODS: Forty eight patients with gallbladder cancer who underwent both EUS and surgery between 1983 and 1998 were included in this study. The accuracy of serial examination by both EUS and angiography in T staging, based on previously established diagnostic criteria, was retrospectively evaluated. First, the presence or absence of subserosal tumor invasion was assessed by EUS alone. Second, in equivocal cases, the depth of tumor invasion was further evaluated by angiographic findings. RESULTS: Twenty-four patients were correctly diagnosed as having other than pT2 cancer by EUS alone. Angiographic findings were reviewed in 19 of the remaining patients, who had pT1, pT2, or a small number of pT3 lesions. The sensitivity, specificity, and overall accuracy in the T staging of pT2 gallbladder cancer was 81.8%, 90.6%, and 88.4%, respectively. CONCLUSIONS: Serial angiographic examination following adequate patient selection by EUS is effective and efficient for the diagnosis of pT2 gallbladder cancer. PMID- 11931534 TI - Protein-losing gastropathy associated with autoimmune disease: successful treatment with prednisolone. AB - We report a patient with protein-losing gastropathy probably associated with autoimmune disease, in whom prednisolone treatment was highly effective. A 45 year-old woman was admitted to our hospital with edema of the face and lower extremities. Blood examination revealed hypoproteinemia, hypoalbuminemia, and hypercholesteremia. Renal biopsy revealed no definite findings of lupus nephritis, including vasculitis. A diagnosis of protein-losing gastropathy was made on the basis of increased alpha1-antitrypsin clearance and 99mTc-labeled human serum albumin scintigram showing abnormal radioactivity in the stomach. Endoscopic gastric biopsies revealed nonspecific inflammation, but marked intramural edema. Based on a slight elevation of antinuclear antibody level, autoimmune disease was suspected to be involved in this patient. Administration of prednisolone, as a diagnostic therapy, alleviated the hypoproteinemia, hypoalbuminemia, and hypercholesteremia. These findings suggest that an autoimmune mechanism could have been involved in this case of protein-losing gastropathy. PMID- 11931536 TI - Primary duodenal adenocarcinoma associated with neurofibromatosis type 1. AB - It is well known that patients with neurofibromatosis have an increased incidence of various neoplasms, most of these being tumors of neural-crest origin, including neurofibromas, leiomyomas, ganglioneuromas, paragangliomas, and carcinoids. However, the occurrence of small-bowel adenocarcinoma is rare. In this article, we report a patient with small-bowel adenocarcinoma, the seventh such reported case. We include a review of the literature and a brief discussion of the implications of this association. We suggest that the association between small-bowel adenocarcinomas and neurofibromatosis may not be fortuitous, and that small-bowel adenocarcinoma should be considered, in particular. in the differential diagnosis of gastrointestinal symptoms in patients with neurofibromatosis. Also, a definite histologic diagnosis must be made, with the employment of special staining techniques, because of the differing prognoses for various cell types. PMID- 11931535 TI - Early appendiceal adenocarcinoma. A review of the literature with special reference to optimal surgical procedures. AB - A rare case of early colonic adenocarcinoma of the appendix confined to the mucosa is reported. The patient remained well 5 years after simple appendectomy. We also review the Japanese literature on early colonic adenocarcinoma of the appendix. Twenty-seven cases of early colonic adenocarcinoma of the appendix, including ours, have been reported in Japan. In 20 of these patients, right hemicolectomy or ileocecal resection was performed. Eighteen patients were available for lymph node evaluation. Lymph nodes were negative for metastasis in 17 of the 18. Only one patient, with poorly differentiated adenocarcinoma invading the submucosa, had lymph node metastases. Our study shows that well differentiated adenocarcinoma invading the submucosa, or adenocarcinoma of any differentiation confined to the mucosa, may be feasibly treated with simple appendectomy. PMID- 11931537 TI - Tumor thrombi in the portal vein system originating from gastrointestinal tract cancer. AB - Intraluminal tumor thrombus in the portal vein (PV) system originating from gastrointestinal (GI) tract cancer is a rare condition. There are two types of such thrombi, one arising indirectly from metastatic liver cancer and the other directly from the primary lesion. We report here three patients with the direct type and two with the indirect type; i.e., a total of five patients with gastric or large intestinal cancer with PV tumor thrombus. In all patients, the primary lesion was surgically resected; in two patients, the tumor thrombus was easily extirpated by direct opening of the PV. It is noteworthy that a patient whose tumor thrombus could not be treated died of cancer with liver failure, caused by expansive growth of the PV tumor thrombus, 4 months after the finding of the PV thrombus. Because PV tumor thrombus may, possibly, determine the patient's length of survival, in addition to causing cancer progression, surgical thrombectomy, combined with resection of the primary cancer and metastatic liver cancer, should be considered for prolongation of survival, if all macroscopic lesions can be controlled and if the tumor thrombus is a synchronous and recent one. PMID- 11931538 TI - Beta-adrenoreceptor agonists for diffuse esophageal spasm. PMID- 11931539 TI - Inadequate health care-seeking behavior of Japanese patients with functional gastrointestinal disorders: a preliminary survey. PMID- 11931540 TI - Rabeprazole: quest for the best PPI. PMID- 11931542 TI - Regulatory networks in prokaryotes: variations on a theme. PMID- 11931543 TI - Regulation of gas vesicle formation in halophilic archaea. AB - The halophilic archaea Halobacterium salinarum and Haloferax mediterranei produce gas vesicles depending on the growth phase and on environmental factors such as light, salt, or oxygen. Fourteen different gvp genes (gvpACNO and gvpDEFGHIJKLM) are involved in their formation, and the regulation of gvp gene expression occurs at the transcriptional and translational level. Haloferax volcanii offers a clean genetic background for the functional analysis of gas vesicle genes by transformation experiments. Such experiments show that the promoter of the gvpA gene encoding the major gas vesicle structural protein is activated by the endogenous basic leucine-zipper protein GvpE. On the other hand, the GvpD protein, which contains a p-loop motif, is involved either directly or indirectly in the repression of the gvpA promoter activity. Eight of the fourteen p-gvp genes (p-gvpAO and p-gvpFGJKLM) enable gas vesicle formation in Hf. volcanii transformants and thus constitute the minimal p-vac region. PMID- 11931541 TI - Does angiotensin II type 1 receptor blockade offer a clinical advantage to cirrhotics with ascites? PMID- 11931544 TI - Sensory transduction to the flagellar motor of Sinorhizobium meliloti. AB - Molecular mechanisms that govern chemotaxis and motility in the nitrogen-fixing soil bacterium, Sinorhizobium meliloti, are distinguished from the well-studied taxis systems of enterobacteria by new features. (i) In addition to six transmembrane chemotaxis receptors, S. meliloti has two cytoplasmic receptor proteins, McpY (methyl-accepting chemotaxis protein) and IcpA (internal chemotaxis protein). (ii) The tactic response is mediated by two response regulators, CheY1 and CheY2, but no phosphatase, CheZ. Phosphorylated CheY2 (CheY2-P) is the main regulator of motor function, whereas CheY1 assumes the role of a 'sink' for phosphate that is shuttled from CheY2-P back to CheA. This phospho-transfer from surplus CheY2-P to CheA to CheY1 replaces CheZ phosphatase. (iii) S. meliloti flagella have a complex structure with three helical ribbons that render the filaments rigid and unable to undergo polymorphic transitions from right- to left-handedness. Flagella rotate only clockwise and their motors can increase and decrease rotary speed. Hence, directional changes of a swimming cell occur during slow-down, when several flagella rotate at different speed. Two novel motility proteins, the periplasmic MotC and the cytoplasmic MotD, are essential for motility and rotary speed variation. A model consistent with these data postulates a MotC-mediated gating of the energizing MotA-MotB proton channels leading to variations in flagellar rotary speed. PMID- 11931545 TI - Regulation of succinoglycan and galactoglucan biosynthesis in Sinorhizobium meliloti. AB - Sinorhizobium meliloti (Rhizobium meliloti) 2011 has the ability to produce the two acidic exopolysaccharides succinoglycan (EPS I) and galactoglucan (EPS II). EPS I is a branched heteropolysaccharide composed of octasaccharide repeating units, whereas EPS II is a linear heteropolysaccharide consisting of disaccharide subunits. The exo-exs and exp gene clusters are involved in the biosynthesis of EPSI and EPSII, respectively. EPSI and EPSII biosynthesis genes are differentially expressed resulting in a complex regulation of EPS production in S. meliloti. The phosphate concentration was identified as an important factor affecting the expression of exp genes. PMID- 11931546 TI - Control of temperature-responsive synthesis of the phytotoxin coronatine in Pseudomonas syringae by the unconventional two-component system CorRPS. AB - The phytopathogen Pseudomonas syringae produces the phytotoxin coronatine (COR) as a major virulence factor. COR and its precursor, coronafacic acid, function as molecular mimics of the plant signaling molecule jasmonate. A 32.8-kb plasmid borne gene cluster mediates COR biosynthesis, which is optimal at 18 degrees C and non-detectable at 28 degrees C, the optimal growth temperature for P. syringae. The thermoregulation is mediated at the transcriptional level by an unconventional two-component regulatory system consisting of a histidine protein kinase, CorS, and two transcriptional activators, CorR and CorP. Dissection of this regulatory triad revealed that CorR binds to its target sequences in a thermoresponsive manner and that its DNA-binding activity is controlled by CorS. A Preliminary model for thermo-sensing by CorS is proposed based on its membrane topology and the analysis of translational fusions of CorS to reporter enzymes at different temperatures. CorP lacks a typical helix-turn-helix motif but possibly functions as a modulator of CorR or CorS activity. The thermoregulation of COR biosynthetic genes is widespread among various COR-producing P. syringae strains. Post-translational processes also contribute to the thermo-responsiveness of COR production. Additionally, COR synthesis in P. syringae is influenced by nutrient availability, rpoN encoding the alternative sigma factor sigma54, and HrpV, a negative regulator of hrp gene expression, suggesting a complex regulatory network governing phytotoxin synthesis. PMID- 11931547 TI - Regulatory factors of Bordetella pertussis affecting virulence gene expression. AB - Most pathogenic bacteria encounter changing growth conditions during their infectious cycle and, accordingly, have to modulate gene expression to enable the efficient colonization of different environments outside or within their host organisms. In Bordetella pertussis the transcription of most virulence factors including several toxins and adhesins is regulated coordinately by the BvgAS two component system. The molecular characterization of the BvgAS system revealed that it belongs to the small group of unorthodox two-component systems applying an obligate multistep phosphorelay. Moreover, despite the coordinated control of the virulence regulon, subtle differences in the regulation of individual virulence genes were observed which led to the identification of sophisticated mechanisms possibly engaged in fine tuning of virulence gene expression. PMID- 11931548 TI - Influence of the leuX-encoded tRNA5(Leu) on the regulation of gene expression in pathogenic Escherichia coli. AB - The leuX gene encoding the minor tRNA5(Leu) is important for the expression of several virulence factors of pathogenic Escherichia coli strains. The differential usage of minor codons to control the expression of specialized genes has been proposed to be a general mechanism of bacteria to regulate gene expression at the posttranscriptional level. The minor codon usage theory foots on the biased codon usage of bacterial genes and the selective availability of tRNA isoacceptors. We aimed at the further investigation of the regulatory role of the tRNA5(Leu) for gene expression in pathogenic E. coli. For this purpose, the molecular mechanism underlying the tRNA5(Leu)-dependent regulation of different virulence-associated genes of pathogenic E. coli as well as the regulation of leuX transcription under various growth conditions were investigated in detail. The global impact of the presence or absence of the leuX encoded tRNA on gene expression of the uropathogenic E. coli strain 536 was studied by proteome analysis. The obtained results argue for a general importance of the tRNA5(Leu) for gene expression of E. coli and the involvement of this tRNA in global regulatory networks. PMID- 11931549 TI - The ciaR/ciaH regulatory network of Streptococcus pneumoniae. AB - New mechanisms for beta-lactam resistance independent on the target penicillin binding proteins were detected in beta-lactam-resistant laboratory mutants of Streptococcus pneumoniae. The link between mutations in the histidine protein kinase CiaH and phenotypic expression of cefotaxime resistance suggests that the cell is able to monitor the integrity of the cell wall and in emergency cases such as during the action of beta-lactams can counteract such danger. At least one ciaH mutation Thr230 > Pro is likely to affect its phosphatase activity resulting in elevated phosphorylation of CiaR, the cognate response regulator, but other CiaH-independent signaling pathways may also result in CiaR phosphorylation. Mutants in CiaH, either alone or in combination with a mutated penicillin-binding protein 2x(PBP2x) fail to develop genetic competence. In all cases complementation of this phenotype was observed upon addition of the competence inducing pheromone peptide CSP, the processed product of the comC gene. This indicates that the cia system is part of a regulatory network that includes another two component system comDE. The DNA binding property of CiaR and ComE were exploited to isolate specifically interacting DNA fragments as a first step to identify genes targeted by individual response regulators. PMID- 11931550 TI - Members of the Fur protein family regulate iron and zinc transport in E. coli and characteristics of the Fur-regulated fhuF protein. AB - The regulator Fur represses with Fe2+ as cofactor iron uptake genes. The fhuF gene reacts very sensitive to minor changes of Fe2+ and Fur. It is assumed that FhuF helps in the mobilisation of iron out of the hydroxamate siderophores transported into the cell. Analysis of the protein revealed an unusual [2Fe-2S] cluster bound to a Cys-Cys-X10-Cys-X2-Cys motif in FhuF. suf genes responsible for the synthesis of the iron sulfur center were identified. The Zur protein shows 27% identity to the Fur protein of E. coli. It regulates as a repressor the high affinity uptake system znuACB. Only two additional Zur binding sites in the promoter region of genes with unknown function were found. Properties of Zur and Fur proteins from different bacteria are compared. PMID- 11931551 TI - Towards an understanding of the molecular mechanisms of stimulus perception and signal transduction by the KdpD/KdpE system of Escherichia coli. AB - The membrane-bound histidine kinase KdpD is a putative turgor sensor that regulates, together with the response regulator KdpE, expression of the kdpFABC operon. This operon encodes the high affinity K+-uptake system KdpFABC of Escherichia coli. Expression of kdpFABC is induced under K+ limiting growth conditions and in response to an osmotic upshift. Various structural features of KdpD and KdpE, which are important for stimulus perception and/or signal transduction were identified and are described here. Furthermore, various studies undertaken to elucidate the nature of the stimulus for KdpD result in a new model for KdpD stimulus perception. According to this, autophosphorylation activity of KdpD is not a result of changes in turgor per se. Instead, various--mainly intracellular parameters--that are related to changes of environmental conditions influence the activities of KdpD. PMID- 11931552 TI - Mechanism of regulation of the bifunctional histidine kinase NtrB in Escherichia coli. AB - NtrB is the bifunctional histidine kinase for nitrogen regulation. Dependent on the availability of nitrogen, it either autophosphorylates and serves as the phosphodonor for its cognate response regulator, NtrC, or, it promotes the rapid dephosphorylation of NtrC-P. The activity of NtrB depends on the interaction of two subdomains within its transmitter domain, the H-domain and the kinase domain. Both phosphotransfer activity and phosphatase activity reside in the H-domain. When separately expressed, this domain acts as a phosphatase. Interaction with the kinase domain results in the inhibition of the phosphatase activity and the phosphorylation of the conserved histidine of the H-domain. PMID- 11931553 TI - Regulation of nitrogen fixation in Klebsiella pneumoniae and Azotobacter vinelandii: NifL, transducing two environmental signals to the nif transcriptional activator NifA. AB - The enzymatic reduction of molecular nitrogen to ammonia requires high amounts of energy, and the presence of oxygen causes the catalyzing nitrogenase complex to be irreversible inactivated. Thus nitrogen-fixing microorganisms tightly control both the synthesis and activity of nitrogenase to avoid the unnecessary consumption of energy. In the free-living diazotrophs Klebsiella pneumoniae and Azotobacter vinelandii, products of the nitrogen fixation nifLA operon regulate transcription of the other nifoperons. NifA activates transcription of nif genes by the alternative form of RNA-polymerase, sigma54-holoenzyme; NifL modulates the activity of the transcriptional activator NifA in response to the presence of combined nitrogen and molecular oxygen. The translationally-coupled synthesis of the two regulatory proteins, in addition to evidence from studies of NifL/NifA complex formation, imply that the inhibition of NifA activity by NifL occurs via direct protein-protein interaction in vivo. The inhibitory function of the negative regulator NifL appears to lie in the C-terminal domain, whereas the N terminal domain binds FAD as a redox-sensitive cofactor, which is required for signal transduction of the internal oxygen status. Recently it was shown, that NifL acts as a redox-sensitive regulatory protein, which modulates NifA activity in response to the redox-state of its FAD cofactor, and allows NifA activity only in the absence of oxygen. In K. pneumoniae, the primary oxygen sensor appears to be Fnr (fumarate nitrate reduction regulator), which is presumed to transduce the signal of anaerobiosis towards NifL by activating the transcription of gene(s) whose product(s) function to relieve NifL inhibition through reduction of the FAD cofactor. In contrast, the reduction of A. vinelandii-NifL appears to occur unspecifically in response to the availability of reducing equivalents in the cell. Nitrogen status of the cells is transduced towards the NifL/NifA regulatory system by the GlnK protein, a paralogue PII-protein, which appears to interact with the NifL/NifA regulatory system via direct protein-protein interaction. It is not currently known whether GlnK interacts with NifL alone or affects the NifL/NifA-complex; moreover the effects appear to be the opposite in K. pneumoniae and A. vinelandii. In addition to these environmental signals, adenine nucleotides also affect the inhibitory function of NifL; in the presence of ATP or ADP the inhibitory effect on NifA activity in vitro is increased. The NifL proteins from the two organisms differ, however, in that stimulation of K. pneumoniae-NifL occurs only when synthesized under nitrogen excess, and is correlated with the ability to hydrolyze ATP. In general, transduction of environmental signals to the nif regulatory system appears to involve a conformational change of NifL or the NifL/NifA complex. However, experimental data suggest that K. pneumoniae and A. vinelandii employ significantly different species-specific mechanisms of signal transduction. PMID- 11931554 TI - Regulation of nitrogen fixation in the phototrophic purple bacterium Rhodobacter capsulatus. AB - In R. capsulatus synthesis and activity of the molybdenum and the alternative nitrogenase is controlled at three levels by the environmental factors ammonium, molybdenum, light, and oxygen. At the first level, transcription of the nifA1, nifA2, and anfA genes--which encode the transcriptional activators of all other nif and anf genes, respectively--is controlled by the Ntr system in dependence on ammonium availability. Mutations in ginB (coding for the signal transduction protein PII) result in significant expression of nifA and anfA in the presence of ammonium. In contrast to GlnB, the PII-paralogue GlnK is not involved in the Ntr signal transduction mechanism. In addition to ammonium control, transcription of anfA is inhibited by traces of molybdenum via the molybdate-dependent repressor proteins MopA and MopB. At the second level of regulation, activity of NifA1, NifA2, and AnfA is inhibited by ammonium in an NtrC-independent manner. This post translational ammonium control of NifA activity is partially released in the absence of GlnK, and completely abolished in a glnB/glnK double mutant. In contrast, AnfA activity is still inhibited by ammonium in the glnB/glnK mutant background. At the third level of regulation, both GlnB and GlnK as well as the (methyl)-ammonium transporter AmtB are involved in ammonium control of the DraT/DraG system, which mediates reversible ADP-ribosylation of both nitrogenase reductases (NifH and AnfH) in response to changes in ammonium availability or light intensity. Most remarkably, in a glnB/glnK double mutant ammonium control of the molybdenum (but not of the alternative) nitrogenase is completely relieved, leading to synthesis of active nitrogenase in the presence of high concentrations of ammonium. PMID- 11931555 TI - Oxygen-regulated expression of genes for pigment binding proteins in Rhodobacter capsulatus. AB - Oxygen is the major external factor affecting the expression of photosynthesis genes in facultatively photosynthetic bacteria. Many investigations over the last years mainly carried out on the closely related species Rhodobacter capsulatus and Rhodobacter sphaeroides have identified a number of proteins involved in the oxygen-regulated signal pathway, in which the RegB/RegA two component system plays a central role. While the RegB/RegA system activates photosynthesis genes under low oxygen tension other proteins like CrtJ and PPBP have a repressing function under high oxygen tension. Additional DNA binding proteins like the integration host factor can modulate the expression of photosynthesis genes. The role of alternative sigma factors in this signal pathway is still unclear. PMID- 11931556 TI - The hydrogen-sensing apparatus in Ralstonia eutropha. AB - Molecular hydrogen is widely used by microorganisms as a source of energy. One of the best studied aerobic hydrogen oxidizers, the beta-proteobacterium Ralstonia eutropha (formerly Alcaligenes eutrophus), harbors two distinct [NiFe] hydrogenases which catalyze the heterolytic cleavage of H2 into 2H+ and 2e-. The genes encoding the hydrogenase subunits are arranged in two large operons together with accessory and regulatory genes involved in hydrogenase biosynthesis. Both operons are transcribed from strong sigma54-dependent promoters. Transcription requires the activation by the HoxA protein, a member of the NtrC family of response regulators. HoxA is only active when H2 is present in the environment. H2 recognition is mediated by a signal transduction complex consisting of the soluble histidine protein kinase HoxJ and a regulatory [NiFe] hydrogenase which acts as an H2 receptor. Biochemical and genetic data suggest that signal transduction between the RH and HoxJ involves an electron transport process. According to our current model the histidine protein kinase HoxJ inactivates HoxA by phosphorylation in the absence H2. This property of the HoxJ HoxA regulator pair is quite different from the behaviour of common two-component regulatory systems. Phosphorylation of HoxA is blocked in the presence of H2 provided the RH can contact HoxJ and transmit the signal to the kinase. Furthermore, hydrogenase gene expression is subject to a global regulatory network in response to the carbon and energy source. HoxA is a major component of this epistatic control the molecular mechanism of which is not yet understood. PMID- 11931558 TI - The molecular biology of formate metabolism in enterobacteria. AB - Formate is the signature compound in the anaerobic metabolism of Escherichia coli and other enterobacteria. Its synthesis and degradation is integrated in a net work of metabolic routes which are elegantly regulated to adjust the carbon flux to the metabolic needs. This review summarises the information on the biochemistry of synthesis and degradation of formate, on the genetics of the members of the regulon and on the mechanism underlying the regulation. PMID- 11931557 TI - Control of FNR function of Escherichia coli by O2 and reducing conditions. AB - The synthesis of the enzymes constituting the electron transport chain of Escherichia coli is controlled by electron acceptors in order to achieve high ATP yields and high metabolic rates as well. High ATP yields (or efficiency) are obtained by the use of electron acceptors for respiration which allow high ATP yields, preferentially O2, and nitrate in the absence of O2. The rate of metabolism is adjusted by use of respiratory isoenzymes which differ in the rate and the efficiency of energy conservation, such as the non-coupling NADH dehydrogenase II (ndh gene) and the coupling NADH dehydrogenase I (nuo genes). By combination of the contrary principles (rate versus efficiency), growth is optimized for growth yields and rates. One of the major transcriptional regulators controlling the switch from aerobic to anaerobic respiration is FNR (fumarate nitrate reductase regulator). FNR is located in the cytoplasm and contains a [4Fe-4S] cluster in the active (anaerobic) state. By reaction with O2 the cluster is converted to a [2Fe-2S] cluster and finally to apoFNR. O2 diffuses into the cytoplasm even at very low O2-tensions (1 microM) where it inactivates [4Fe-4S] x FNR. The formation of [4Fe-4S] x FNR from apoFNR can use glutathione as a reducing agent in vitro. This process could also be important for the reductive activation of FNR in vivo. A model for the control of the functional state of FNR by O2 and glutathione is discussed. According to this model the functional state of FNR is determined by a (rapid) inactivation of FNR by O2, and a slow (constant) reactivation with glutathione as the reducing agent. PMID- 11931559 TI - Nitric oxide signaling and NO dependent transcriptional control in bacterial denitrification by members of the FNR-CRP regulator family. AB - Bacterial denitrification transforms nitrate to dinitrogen. The process is expressed facultatively in response to environmental conditions. Around 50 components make up the denitrification apparatus and its assembly pathways. We are beginning to understand how exogenous signals provided by oxygen and N oxides are processed for activating the underlying gene programs. Key signals are provided by nitrate, nitric oxide, and a low oxygen tension. In the genus Pseudomonas the nitrate signal is processed by a two component regulatory system which activates the nar operon encoding respiratory nitrate reductase. Nitric oxide is not only an essential respiratory substrate of the denitrifying cell but constitutes in nanomolar concentrations also a key signal for the expression of nitrite reductase and NO reductase which control cellular NO homeostasis. The signal pathway in the genera Pseudomonas, Paracoccus and Rhodobacter involves regulators of the FNR family of transcription factors, which cluster phylogenetically in a separate subgroup. In contrast, Ralstonia eutropha requires a sigma-54 dependent regulator of the NtrC family for the expression of its quinol-dependent NO reductase. Important questions are directed currently at the mechanism(s) of activation of these transcription factors by NO, and avoidance of crosstalk with FNR factors at target promoters operating with identical recognition motifs. PMID- 11931560 TI - Regulation of heme biosynthesis in non-phototrophic bacteria. AB - The biosynthesis of tetrapyrroles like hemes and chlorophylls is essential for most living organisms. In bacteria hemes are integral parts of energy conserving electron transport chains and cofactors of various enzymes. Changes of environmental conditions usually lead to an adaption of the bacterial energy metabolism and often coincide with significant changes of cellular heme levels. This review focuses on the known regulatory mechanisms in non-phototrophic bacteria involved in the control of the formation of the heme biosynthetic apparatus. Species specific differences in the mode of energy generation result in various regulatory strategies. Focusing on the well investigated bacteria Bacillus subtilis, Escherichia coli, Pseudomonas aeruginosa, and Salmonella typhimurium the involved environmental stimuli, employed transcriptional regulators and promoter structures as well as the role of protein stability are described. Broad variations of the used regulatory principles were observed. PMID- 11931562 TI - Network regulation of the Escherichia coli maltose system. AB - The genes of the Escherichia coli maltose regulon are controlled by MalT, the specific transcriptional activator which, together with the inducer maltotriose and ATP, is essential for mal gene transcription. Network regulation in this system affects the function of MalT and occurs on two levels. The first concerns the expression of malT. It has long been known that malT is under catabolite repression and thus under the control of the cAMP/CAP complex. We found that, in addition, the global regulator Mlc is a repressor for malT transcription. The repressor activity of Mlc is controlled by the transport status of the glucose specific enzyme EIICB of the PTS that causes sequestration (and inactivation as a repressor) of Mlc when glucose is transported. The second level of MalT regulation affects its activity. MalT is activated by maltotriose which is not only formed when the cells are growing on any maltodextrin but also, in low amounts, endogenously when the cells grow on non-maltodextrin carbon sources. Thus, cellular metabolism, for instance degradation of galactose or trehalose, can cause mal gene induction. It was found that unphosphorylated internal glucose takes part in endogenous maltodextrin biosynthesis and is therefore a key element in endogenous mal gene expression. In addition to the maltotriose-dependent activation, MalT can interact with three different enzymes that lead to its inactivation as a transcriptional activator. The first is MaIK, the energy transducing ABC subunit of the maltodextrin transport system. Transport controls the interaction of MalK and MalT thus affecting gene expression. The second enzyme is MalY, a pyridoxal phosphate containing enzyme exhibiting cystathionase activity. The crystal structure of MalY was established and mutations in MalY that reduce mal gene repression map in a hydrophobic MalT interaction patch on the surface of the enzyme. The last enzyme is a soluble esterase of as yet unknown function. When overproduced, this enzyme specifically reduces mal gene expression and affects the activity of MalT in an in vitro transcription assay. PMID- 11931561 TI - Transcriptional regulation of solventogenesis in Clostridium acetobutylicum. AB - Solvent synthesis in Clostridium acetobutylicum is induced in concert with sporulation to counteract the dangerous effects of produced butyric and acetic acids and to provide the cell with sufficient time to complete endospore formation. Cardinal transcription units for butanol and acetone production are the sol and adc operons encoding butyraldehyde/butanol dehydrogenase and coenzyme A transferase as well as acetoacetate decarboxylase. Induction is achieved by a decreased level of DNA supercoiling and the transcription factor Spo0A, possibly in cooperation with other regulatory proteins. A number of other operons is also turned on during this metabolic switch, whose physiological relevance, however, is only partly understood. The recent completion of C. acetobutylicum genome sequencing will pave the way for transcriptional profiling and thus allow comprehension of the coherent regulatory networks of solventogenesis and sporulation. PMID- 11931563 TI - Carbon catabolite repression by the catabolite control protein CcpA in Staphylococcus xylosus. AB - Carbon catabolic repression (CR) by the catabolite control protein CcpA has been analyzed in Staphylococcus xylosus. Genes encoding components needed to utilize lactose, sucrose, and maltose were found to be repressed by CcpA. In addition, the ccpA gene is under negative autogenous control. Among several tested sugars, glucose caused strongest CcpA-dependent repression. Glucose can enter S. xylosus in nonphosphorylated form via the glucose uptake protein GlcU. Internal glucose is then phosphorylated by the glucose kinase GlkA. Alternatively, glucose can be transported and concomitantly phosphorylated by glucose-specific permease(s) of the phosphotransferase system (PTS). S. xylosus mutant strains deficient in GlcU or GlkA showed partial relief of glucose-specific, CcpA-dependent repression. Likewise, blocking PTS activity completely by inactivation of the gene encoding the general PTS protein enzyme I resulted in diminished glucose-mediated repression. Thus, both glucose entry routes contribute to glucose-specific CR in S. xylosus. The sugar transport activity of the PTS is not required to trigger glucose-specific repression. The phosphocarrier protein HPr however, is absolutely essential for CcpA activity. Inactivation of the HPr gene led to a complete loss of CR. Repression is also abolished upon inactivation of the HPr kinase gene or by replacing serine at position 46 of HPr by alanine. These results clearly show that HPr kinase provides the signal, seryl-phosphorylated HPr, to activate CcpA in S. xylosus. PMID- 11931564 TI - CcpA-independent carbon catabolite repression in Bacillus subtilis. AB - The past decade has witnessed an exiting unveiling of numerous molecular mechanisms that characterize signal transduction by protein-protein interaction. The recent findings encouraged an increasing effort to understand the sequential metabolism of different sugars available as energy sources at the same time. It seems probable that at least three principle mechanisms which act together or separately, mediate carbon catabolite repression (CCR) depending on the system which is under metabolic control: i) by the main signal transducing chain via the ATP-dependent HPr-kinase, HPr(Ser46-P) or alternatively Crh via the central component CcpA and its interaction with cre, ii) by signals sensed from the specific regulators directly or via phosphorylation by HPr, iii) by inducer exclusion based on the concurrence of the enzyme IIA(Glc) domain of the glucose permease, and other PTS-dependent permeases composed only of the B and C domains and lacking the enzyme IIA domain. PMID- 11931565 TI - Structure-function relationship and regulation of two Bacillus subtilis DNA binding proteins, HBsu and AbrB. AB - Microorganisms use a number of small basic proteins for organization and compaction of their DNA. By their interaction with the genome, these proteins do have a profound effect on gene expression, growth behavior, and viability. It has to be distinguished between indirect effects as a consequence of the state of chromosome condensation and relaxation that influence the rate of RNA polymerase action as represented by the histone-like proteins, and direct effects by specific binding of proteins to defined DNA segments predominantly located around promoter sequences. This latter class is represented by the transition-state regulators that are involved in integrating various global stimuli and orchestrating expression of the genes under their regulation for a better adaptation to changes in growth rate. In this article we will focus on two different but abundant DNA binding proteins of the gram-positive model organism Bacillus subtilis, the histone-like HBsu as a member of the unspecific and the transition state regulator AbrB as a member of specific classes of DNA binding proteins. PMID- 11931566 TI - Regulation of ribosomal RNA synthesis in E. coli: effects of the global regulator guanosine tetraphosphate (ppGpp). AB - The global regulatory nucleotides (p)ppGpp are major effectors for the control of ribosomal RNA in bacteria. The effector molecules accumulate to different cellular levels at amino acid deprivation or during different growth rates. They change the activity of RNA polymerase to transcribe from sensitive promoters (e.g. ribosomal RNA promoters). Sensitive promoters are characterized by a GC rich discriminator element in addition to further structural requirements not completely understood. ppGpp must also be regarded as a mediator for growth rate control although it appears that ppGpp-independent regulatory mechanisms exist. Inhibition occurs at various steps during initiation but also during elongation where RNA polymerase pausing is observed. From the existing data a mechanistic model for the action of ppGpp is suggested considering structural details of RNA polymerase obtained at high resolution. PMID- 11931567 TI - Recent insights into the general stress response regulatory network in Escherichia coli. AB - Many bacterial species exhibit a general stress response that can be induced by numerous very different stress conditions and, phenotypically, renders the cells broadly stress resistant. In Escherichia coli, this response is dependent on the sigmaS (RpoS) subunit of RNA polymerase. SigmaS is a close relative of the vegetative sigma factor sigma70 (RpoD) and recognizes very similar promoter sequences. In recent years, significant progress has been made with respect to elucidating (i) the molecular mechanisms that control the cellular sigmaS level, which include translational regulation as well as intricate control of sigmaS proteolysis, and (ii) the molecular function of sigmaS as a transcription initiation factor, where a number of sigmaS-dependent promoters have been studied in great detail, and the mechanisms that generate sigmaS selectivity are now becoming apparent. PMID- 11931568 TI - Chemical genomics: a systematic approach in biological research and drug discovery. AB - The knowledge of complete sequences of different organisms is dramatically changing the landscape of biological research and pharmaceutical development. We are experiencing a transition from a trial-and-error approach in traditional biological research and natural product drug discovery to a systematic operation in genomics and target-specific drug design and selection. Small, cell-permeable and target-specific chemical ligands are particularly useful in systematic genomic approaches to study biological questions. On the other hand, genomic sequence information, comparative and structural genomics, when combined with the cutting edge technologies in synthetic chemistry and ligand screening/identification, provide a powerful way to produce target-specific and/or function-specific chemical ligands and drugs. Chemical genomics or chemogenomics is a new term that describes the development of target-specific chemical ligands and the use of such chemical ligands to globally study gene and protein functions. We anticipate that chemical genomics plays a critical role in the genomic age of biological research and drug discovery. PMID- 11931569 TI - An overview of toxicogenomics. AB - Toxicogenomics is a rapidly developing discipline that promises to aid scientists in understanding the molecular and cellular effects of chemicals in biological systems. This field encompasses global assessment of biological effects using technologies such as DNA microarrays or high throughput NMR and protein expression analysis. This review provides an overview of advancing multiple approaches (genomic, proteomic, metabonomic) that may extend our understanding of toxicology and highlights the importance of coupling such approaches with classical toxicity studies. PMID- 11931570 TI - Normalizing DNA microarray data. AB - DNA microarrays are a powerful tool to investigate differential gene expression for thousands of genes simultaneously. Although DNA microarrays have been widely used to understand the critical events underlying growth, development, homeostasis, behavior and the onset of disease, the management of the resulting data has received little attention. Presently, the fluorescent dyes Cy3 and Cy5 are most often used to prepare labeled cDNA for microarray hybridizations. Raw microarray data are image files that have to be transformed into gene expression formats--a process that requires data manipulation due to systematic variations which may be attributed to differences in the physical and chemical dye applications is to identify differences in transcript levels calculated from fluorescence ratios it is necessary to normalize fluorescence signals to compensate for systematic variations. Here, we will review current normalization strategies applied to cDNA microarrays and discuss their limits. We will show that experimental design determines normalization success. PMID- 11931571 TI - Colonization models and initial genetic diversity in the Americas. AB - The mode and tempo of colonization of the Americas established the initial pattern of continental genetic diversity. Despite a long history of study, the process of settlement remains controversial in terms of date, rate, and pattern. While there is agreement that Asia was the source population, several different models have been proposed for the colonization process. A classic model postulates a rapid spread of population ("blitzkrieg") from a small band of hunters entering through the corridor between the continental ice sheets circa 11,000 years B.P. Colonization occurred as a wave of expansion across the land masses of North and South America. An alternative model envisions the original colonists initially limiting settlement to the coastline, using boats, and entering the Americas at an earlier date, circa 13,500 B.P. Range expansion along this linear habitat from North to South America could be rapid without requiring population saturation of entire continental regions. These models have markedly different implications for genetic variation among Native Americans. The blitzkrieg colonization process would have generated multiple founder effects leading to extreme loss of genetic variation. Computer simulation of this model shows nearly complete fixation in 30 generations. Simulation of the coastal model, on the other hand, requires less extreme demographic assumptions and maintains substantial genetic variability after 100 generations. Although with the coastal model continental interiors are occupied less rapidly than with the blitzkrieg model, the coastal model allows earlier entry and rapid expansion to the southern limits of the hemisphere. PMID- 11931572 TI - Segregation analysis of blood pressure and body mass index in a rural US community. AB - To assess evidence for a gene with large effect on systolic blood pressure (SBP), diastolic blood pressure (DBP), and body mass index (BMI), we conducted segregation analyses on 261 nuclear families collected from a rural Caucasian community in Michigan. The families were ascertained through a hypertensive proband. Each phenotype was adjusted for significant covariate effects (e.g., gender and age). We used class D regressive models to conduct the segregation analyses. Our analysis results support the segregation of a major gene for BMI, but not for SBP or DBP. A recessive locus effect provided the best explanation for BMI where approximately 43% of the variance of BMI was due to this gene. PMID- 11931573 TI - The impact of labor-saving technology on first birth intervals in rural Ethiopia. AB - Across the developing world labor-saving technologies introduce considerable savings in the time and energy that women allocate to work. Hormonal studies on natural fertility populations indicate that such a reduction in energetic expenditure (rather than improved nutritional status alone) can lead to increased ovarian function. Other qualitative studies have highlighted a link between labor saving technology and behavioral changes affecting subsequent age at marriage, which may affect fertility. This biodemographic study was designed to investigate whether these physiological and behavioral changes affect fertility at a population level by focusing on a recent water development scheme in Southern Ethiopia. The demographic consequences of a reduction in women's workload following the installation of water points, specifically the variation in length of first birth interval (time lapsed between marriage and first birth), are investigated. First birth interval length is closely associated with lifetime fertility in populations that do not practice contraception, longer intervals being associated with lower fertility. Using life tables and multivariate hazard modeling techniques a number of significant predictors of first birth interval length are identified. Covariates such as age at marriage, season of marriage, village ecology, and access to improved water supply have significant effects on variation in first birth intervals. When entered into models as a time-varying covariate, access to a water tap stand is associated with an immediate reduction in length of first birth intervals. PMID- 11931574 TI - The relationship among apolipoprotein(a) polymorphisms, the low-density lipoprotein receptor-related protein, and the very low density lipoprotein receptor genes, and plasma lipoprotein(A) concentration in the Czech population. AB - Increased plasma concentration of lipoprotein(a) [Lp(a)] is an established independent risk factor for coronary artery disease (CAD), which is strongly genetically determined. This study was designed to investigate the relationship between the K-IV and (TTTTA)n apolipoprotein(a) [apo(a), protein; APOA, gene] polymorphisms, as well as the C766T low-density lipoprotein receptor-related protein (LRP) and the (CGG)n very low density lipoprotein receptor (VLDLR) polymorphisms on the one hand, and plasma Lp(a) levels in Czech subjects who underwent coronary angiography on the other hand. The lengths of the alleles of the APOA K-IV and (TTTTA)n polymorphisms were strongly inversely correlated with plasma Lp(a) levels in univariate analysis (r = -0.41, p < 10(-4) and r = -0.20, p < 0.01, respectively). Multivariate analysis revealed significant associations between the APOA polymorphisms studied and plasma Lp(a) levels in subjects expressing only one APOA K-IV allele (p < 10(-6) for K-IV and p < 0.001 for TTTTA). In subjects expressing both APOA K-IV alleles, the multivariate analysis revealed that only the APOA K-IV alleles were inversely correlated with plasma Lp(a) levels (p < 0.001). Associations between both the LRP and VLDLR gene polymorphisms and plasma Lp(a) levels were only of borderline significance (p < 0.06 and p < 0.07, respectively) and were not confirmed in multivariate analysis. In conclusion, both APOA length polymorphisms significantly influenced plasma Lp(a) concentration in the Czech population studied, and this circumstance could explain the association in this population observed earlier between APOA (TTTTA)n polymorphism and CAD (Benes et al. 2000). Only a minor role in the regulation of plasma Lp(a) levels is suggested for the C766T LRP and the (CGG)n VLDLR polymorphisms. PMID- 11931575 TI - Allele frequencies of apolipoprotein E gene polymorphisms in the protein coding region and promoter region (-491A/T) in a healthy Norwegian population. AB - This study examines the distribution of apolipoprotein E (APOE) alleles in a population of healthy male and female Norwegians (n = 798) below the age of 40. The -491A/T polymorphism of the promoter region of the APOE gene was also examined. A seminested polymerase chain reaction was applied in the genotyping. The results showed that the E3 allele had the highest frequency (0.744), followed by E4 (0.198) and E2 (0.058). The APOE frequencies found in this study differ significantly from those obtained in earlier Norwegian APOE phenotypings. The allele frequencies in the -491 site of the promoter region were 0.845 for the A allele and 0.155 for the T allele. The genotype frequency was highest for AA (0.707), followed by AT (0.277) and TT (0.016). Moreover, the A allele was in linkage disequilibrium to E4. PMID- 11931576 TI - Characterization of two New York City Jewish populations at six short tandem repeat loci. AB - The Hasidic and non-Hasidic Jewish communities of New York City represent two subpopulations with long-documented histories of restrictive marriage patterns and a high degree of endogamy. As part of a continuing study into their genetic structure, allele frequencies were determined for the six tetrameric short tandem repeat (STR) loci: FESFPS, F13AO1, vWA, CSF1PO, TPOX, and THO1. All loci were tested for Hardy-Weinberg equilibrium (HWE) by three tests: chi-square analysis, Monte Carlo chi-square analysis. and the exact test. The non-Hasidic population failed to meet HWE at the F13A01, FESFPS, and CSF1PO loci by all three tests. The Hasidic population also failed to meet HWE at the same loci by some of the tests. Comparison of the Hasidic to the non-Hasidic population using an R x C contingency table demonstrated a similarity at only the vWA locus. Significant differences exist when comparing the two Jewish populations to a reference Caucasian population. PMID- 11931577 TI - Characterization of apolipoprotein E genetic variations in Taiwanese: association with coronary heart disease and plasma lipid levels. AB - Apolipoprotein E (apoE, protein; APOE, gene) is important in lipoprotein metabolism. Three isoforms, apoE2 (Cys112 Cys158), apoE3 (Cys112 Arg158), and apoE4 (Arg112 Arg158), are present in the general population. This report investigates the frequency distribution of apoE isoforms and the association of APOE genotypes with plasma lipid profile and coronary heart disease (CHD) in a population of Taiwan. ApoE isoforms were determined genetically by polymerase chain reaction and HhaI restriction enzyme digestion in control and coronary heart disease (CHD) patients. Plasma lipid and lipoprotein concentrations were also determined. The control group exhibited frequencies of 84.6% APOE3, 7.9% APOE4, 7.5% APOE2, 70.6% APOE3E3, 14.4% APOE3E4, 13.6% APOE2E3, and 1.4% APOE2E4. Comparable frequencies were observed in the CHD group. In both APOE2 carrier and APOE3E3 groups, the CHD patients expressed abnormal lipid profiles while the control group expressed normal lipid profiles. The APOE4 carriers, however, expressed abnormal lipid profiles in both normal control and CHD groups. Extremely high apoE levels in the hypertriglyceridemic group (TG > 400 mg/dL) seemed to be undesirable and were often observed in CHD patients. PMID- 11931578 TI - Patterns of genetic diversity at the nine forensically approved STR loci in the Indian populations. AB - Genetic diversity at the nine short tandem repeat (STR) loci, which are universally approved and widely used for forensic investigations, has been studied among nine Indian populations with diverse ethnic, linguistic, and geographic backgrounds. The nine STR loci were profiled on 902 individuals using fluorescent detection methods on an ABI377 System, with the aid of an Amp-F1 Profiler Plus Kit. The studied populations include two upper castes, Brahmin and Kayastha; a tribe, Garo, from West Bengal; a Hindu caste, Meitei, with historical links to Bengal Brahmins; a migrant group of Muslims; three tribal groups, Naga, Kuki and Hmar, from Manipur in northeast India; and a middle-ranking caste, Golla, who are seminomadic herders from Andhra Pradesh. Gene diversity analysis suggests that the average heterozygosity is uniformly high (>0.8) in the studied populations, with the coefficient of gene differentiation at 0.050 +/- 0.0054. Both neighbor-joining (NJ) and unweighted pair group method with arithmetic mean (UPGMA) trees based on DA distances bring out distinct clusters that are consistent with ethnic, linguistic, and/or geographic backgrounds of the populations. The fit of the Harpending and Ward model of regression of average heterozygosity on the gene frequency centroid is found to be good, and the observed outliers are consistent with the population structure and history of the studied populations. Our study suggests that the nine STR loci, used so far mostly for forensic investigations, can be used fruitfully for microevolutionary studies as well, and for reconstructing the phylogenetic history of human populations, at least at the local level. PMID- 11931579 TI - Structuring the genetic heterogeneity of the Basque population: a view from classical polymorphisms. AB - In this study we analyze 18 classical polymorphisms (ABO, Rh, MNSs, Lewis, P, Duffy, Kell, ADA, ESD, PGM1, PGD, AK1, ACP1, GLO1, HP, GC, TF, and PI) in over 2000 autochthonous individuals from 14 natural districts in three provinces of the Basque Country (Alava, Guipuzcoa, and Biscay). Heterogeneity analysis via the chi2 test and a calculation of F(ST) indicate that there is significant genetic heterogeneity between the Basque districts. The R matrix informs us that this heterogeneity is not significantly concentrated in a single district or in the districts of a single province, but is rather distributed among several districts belonging to the three provinces analyzed. We undertake to assess the influence of various historical, geographical, and cultural factors on the genetic structure of the Basque population. Analysis suggests that allele distribution is geographically patterned in the Basque Country. The gradient distributions observed in the case of some alleles (ABO*O, RH*cDE, RH*cde, MNS*MS, and ACP1*C) on the basis of Moran's autocorrelation coefficient I, along with the influence of the two main travel routes through the Basque Country (western route through Bilbao and eastern route through Vitoria), suggest that the gene flow tends toward the coast. As regards other factors considered (administrative division, repopulation processes, linguistic heterogeneity, and north vs. south cultural heterogeneity), we detected only a certain influence exerted by an old tribal differentiation (2000 B.P.), which would diminish with the passage of time. PMID- 11931580 TI - Misclassification, correlation, and cause of death studies. AB - Relatives may tend to die of the same cause. A model is proposed specifying the degree of this association, simultaneously allowing for misclassification of cause of death. These mistakes usually degrade any existing correlation structure, the effects being marked when error rates reach between 20% to 30%. It is then that the search for associations becomes very difficult indeed, and results misleading. This is an example of a wider malaise that can afflict analysis of association, and the developments here apply to two-by-two contingency tables in general. PMID- 11931581 TI - Ethnic-immigrant differentials in health behaviors, morbidity, and cause-specific mortality in the United States: an analysis of two national data bases. AB - This study examines the extent to which various ethnic-immigrant and US-born groups differ in their risks of all-cause and cause-specific mortality, morbidity, and health behaviors. Using data from the National Longitudinal Mortality Study, 1979-1989, we estimated, for major US racial and ethnic groups, mortality risks of immigrants relative to those of the US-born. The Cox regression model was used to adjust mortality differentials by age, sex, marital status, rural/urban residence, education, and family income. Logistic regression was fitted to the National Health Interview Survey data to determine whether health status and behaviors vary among ethnic-immigrant groups and by length of US residence. Compared with US-born whites of equivalent socioeconomic and demographic background, foreign-born blacks, Hispanics, and Asians/Pacific Islanders (APIs), US-born APIs, US-born Hispanics, and foreign-born whites had, respectively, 48%, 45%, 43%, 32%, 26%, and 16% lower mortality risks. While American Indians did not differ significantly from US-born whites, US-born blacks had an 8% higher mortality risk. Black and Hispanic immigrants experienced, respectively, 52% and 26% lower mortality risks than their US-born counterparts. Considerable differentials were also found in mortality for cancer, cardiovascular, respiratory, infectious disease, and injury, and in morbidity and health behaviors, with API and Hispanic immigrants generally experiencing the lowest risks. Consistent with the acculturation hypothesis, immigrants' risks of smoking, obesity, hypertension, and chronic condition, although substantially lower than those for the US-born, increased with increasing length of US residence. Given the substantial nativity differences in health status and mortality, future waves of immigrants of diverse ethnic and cultural backgrounds will likely have a sizeable impact on the overall health, disease, and mortality patterns in the United States. PMID- 11931582 TI - Purification of angularin, a novel antifungal peptide from adzuki beans. AB - An antifungal peptide was isolated from the adzuki bean with a procedure involving affinity chromatography on Affi-gel blue gel and ion exchange chromatography on CM-Sepharose. The protein designated angularin was adsorbed on both types of chromatographic media and possessed a molecular weight of 8 kDa. Angularin exhibited antifungal activity against a variety of fungal species including Mycospharella arachidiocola and Botrytis cinerea. It inhibited mycelial growth in B. cinerea with an IC50 of 14.3 microM. Fusarium oxysporum and Rhizoctonia solani were not inhibited. Angularin demonstrated inhibitory activity on translation in the rabbit reticulocyte lysate system (IC50 = 8.0 microM) but did not affect proliferation of splenocytes. The activity of HIV-1 reverse transcriptase was inhibited in the presence of angularin. Its N-terminal sequence was GEPGQKE. PMID- 11931583 TI - Peptide based vaccine design: synthesis and immunological characterization of branched polypeptide conjugates comprising the 276-284 immunodominant epitope of HSV-1 glycoprotein D. AB - The importance of the length and conjugation site of a protective epitope peptide (276SALLEDPVG284) from glycoprotein D of herpes simplex virus in branched polypeptide conjugates has been investigated. A new set of peptides, with a single attachment site and truncated sequences, was prepared. The immunogenicity of conjugates and the specificity of antibody responses elicited were investigated in BALB/c, C57/B1/6 and CBA mice. It was found that the covalent coupling of the peptide comprising the 276-284 sequence of gD through its Asp residue at position 281 did not influence the immunogenic properties of the epitope, while involvement of the side chain of Glu at position 280 almost completely abolished immunogenicity. These results clearly indicated that the conjugation site of the epitope peptide influenced the intensity and specificity of antibody responses. Comparison of the immunological properties of conjugates containing truncated gD peptides revealed the presence of two epitopes within the 276-284 region. One of the proposed epitopes is situated at the N-terminal (276 281) region, while the other is located at the C-terminal end of the sequence (279-284). Binding data demonstrated that some of the peptides comprising these epitopes induced gD-specific responses in their conjugated form and also elicited an immune response that conferred protection against lethal HSV-1 infection. The correlation of peptide- and gD-specific antibody responses with the protective effect of the immune response is discussed. PMID- 11931584 TI - The production of anti-hexapeptide antibodies which recognize the S7, L6 and L13 ribosomal proteins of Escherichia coli. AB - Here we report the synthesis of the N-terminal hexapeptide H-Pro-Arg-Arg-Arg-Val Ile-OH of the E. coli ribosomal protein S7. the C-terminal hexapeptide H-Lys-Glu Ala-Lys-Lys-Lys-OH of L6 and the C-terminal hexapeptide H-Pro-Gln-Val-Leu-Asp-Ile OH of L13. All peptides were prepared by SPPS following the Fmoc-strategy, using DIC/HOBt and/or HBTU as coupling reagents and 2-chlorotrityl chloride resin as the solid support. The carrier linked synthetic peptides were injected into rabbits and elicited an anti-peptide response. These anti-hexapeptide antibodies were found to recognize the corresponding peptides and proteins. PMID- 11931585 TI - The effect of C-terminal helix stabilization on specific DNA binding by monomeric GCN4 peptides. AB - DNA binding by a 29-residue, monomeric, GCN4 basic region peptide, GCN4br, as well as by peptide br-C, a monomeric basic-region analogue that is helix stabilized at its C-terminal end by a Lys25. Asp29 side-chain lactam-bridged alanine-rich sequence, was studied at 25 C in an aqueous buffer containing 100 mm NaCl. Mixing of both peptides with duplex DNA containing the cAMP-responsive element (CRE) was accompanied by significant helix stabilization in the peptides, whereas mixing of the peptides with duplex DNA containing a scrambled CRE site was not. Peptide NBD-br-C was synthesized as a fluorescent probe to evaluate these peptide-DNA interactions further. Quantitative analysis of the fluorescence quenching of peptide NBD-br-C by CRE half-site DNA indicated the formation of a 1:1 complex with a dissociation constant of 1.41 +/- 0.22 microm. Competitive displacement fluorescence assays of CRE half-site binding gave dissociation constants of 0.65 +/- 0.09 microm for peptide br-C and 3.9 +/- 0.5 microM for GCN4br, which corresponds to a free energy difference of 1.1 kcal/mol that is attributed to the helix stabilization achieved in peptide br-C. This result indicates that helix initiation by the alpha-helical leucine zipper dimerization motif in native bzip proteins, such as GCN4, contributes significantly to the affinity of basic region peptides for their recognition sites on DNA. Our fluorescence assay should also prove useful for determining dissociation constants for CRE binding by other GCN4 basic region analogues under equilibrium conditions and physiological salt concentrations. PMID- 11931586 TI - Intestinal transport of the lactokinin Ala-Leu-Pro-Met-His-Ile-Arg through a Caco 2 Bbe monolayer. AB - ACE inhibitory peptides are biologically active peptides that play a role in blood pressure regulation. When derived from food proteins during food processing or gastrointestinal digestion, these peptides could function as efficient agents in treating and preventing hypertension. However, in order to exert an antihypertensive effect by inhibition of the ACE enzyme, they have to reach the bloodstream intact. The aim of this research was to assess if the known ACE inhibitory peptide Ala-Leu-Pro-Met-His-Ile-Arg, derived from a tryptic digest of beta-lactoglobulin, could be absorbed through a Caco-2 Bbe cell monolayer in an Ussing chamber and reach the serosal side undegraded. Samples of the mucosal compartment showed high ACE inhibitory activity. No or only little ACE inhibitory activity was detected in the serosal compartment. However, when the serosal sample was concentrated three-fold, a substantial ACE inhibitory activity was registered. Concomitantly, HPLC and MS clearly showed the presence of Ala-Leu-Pro Met-His-Ile-Arg in the mucosal compartment, whereas in the serosal compartment only MS was able to detect the heptapeptide. In conclusion. under the observed experimental conditions, the ACE inhibitory peptide Ala-Leu-Pro-Met-His-Ile-Arg was transported intact through the Caco-2 Bbe monolayer, but in concentrations too low to exert an ACE inhibitory activity. PMID- 11931587 TI - Paroxetine in adolescent major depression. PMID- 11931588 TI - Treatment of preschool OCD. PMID- 11931589 TI - Paroxetine in adolescent major depression. PMID- 11931590 TI - Adderall and seizures. PMID- 11931591 TI - Risperidone-induced retrograde ejaculation. PMID- 11931592 TI - Lining up for children's mental health services: a tool for prioritizing waiting lists. AB - OBJECTIVE: The Western Canada Waiting List Project (WCWL) is a federally funded partnership of 19 health-related organizations that was created to develop tools to manage waiting lists for five types of health services. METHOD: The children's mental health (CMH) panel developed and tested a set of standardized clinical criteria for setting priorities among patients awaiting CMH services. The criteria were applied to 817 patients by 92 mental health professionals in three western provinces. Regression analysis was used to determine the set of criteria weights that collectively best predicted clinicians' global urgency ratings. To assess reliability, raters used the criteria to score six standardized "paper cases." RESULTS: The criteria accounted for about 40% of the observed variance in overall urgency ratings (R2 = 41.7%). The panel modified the criteria on the basis of the initial empirical work. Reliability assessment of the revised tool indicated that half of the items had excellent or fair/good interrater agreement; test-retest reliability was good. CONCLUSIONS: Priority criteria were able to capture clinicians' judgments of relative urgency in the CMH setting. A number of operational challenges remain with the use of priority criteria for scheduling CMH services. Further development and testing of the tool appear warranted. PMID- 11931593 TI - Case-control study of attention-deficit hyperactivity disorder and maternal smoking, alcohol use, and drug use during pregnancy. AB - OBJECTIVE: To address the putative association between attention-deficit hyperactivity disorder (ADHD) and prenatal exposure to maternal cigarette smoking, drugs of abuse, and alcohol attending to potential confounding by familial ADHD, maternal depression, conduct disorder, and indicators of social adversity in the environment. METHOD: A retrospective, hospital-based, case control study was conducted with 280 ADHD cases and 242 non-ADHD controls of both genders. The case and control children and their relatives were systematically assessed with structured diagnostic interviews. Logistic regression analysis was used to determine the adjusted effect of prenatal exposure to substance use and ADHD. RESULTS: ADHD cases were 2.1 times (95% confidence interval = 1.1-4.1;p = .02) more likely to have been exposed to cigarettes and 2.5 times (95% confidence interval = 1.1-5.5; p = .03) more likely to have been exposed to alcohol in utero than were the non-ADHD control subjects. Adjustment by familial psychopathology, Rutter's indicators of social adversity, and comorbid conduct disorder did not account for the effect of prenatal exposure to alcohol or the products of cigarettes. CONCLUSIONS: ADHD may be an additional deleterious outcome associated with prenatal exposure to alcohol independently of the association between prenatal exposure to nicotine and smoke products and other familial risk factors for the disorder. PMID- 11931594 TI - Development of substance dependence in two delinquency subgroups and nondelinquents from a male twin sample. AB - OBJECTIVE: The effect of delinquency subtype on the development of substance dependence symptoms was examined. It was proposed that early-onset delinquents possess characteristics that increase their likelihood of developing substance dependence problems earlier and more rapidly than late-onset delinquents and nondelinquents. METHOD: The development of alcohol, nicotine, and cannabis dependence symptoms (DSM-III-R) was examined over a 6-year period of adolescence (age 11-17) among 36 early-onset delinquent, 86 late-onset delinquent, and 25 nondelinquent boys from a large epidemiological twin sample. Multilevel/random coefficients models were used to compare groups on the rate of growth in number of symptoms over time. RESULTS: As expected, early-onset delinquents showed an earlier onset and a faster rate of increase in the number of cannabis and nicotine dependence symptoms than late-onset delinquents and controls. Both delinquent groups had a more rapid increase in alcohol dependence symptoms than controls. CONCLUSIONS: The data showed that early-onset delinquency is associated with earlier onset of substance use disorder symptoms and more rapid acceleration of problems with drugs than late-onset delinquency. Treatments for boys with early-onset delinquency should account for their increased risk for drug use problems in adolescence and the potential effects of those problems on the course of antisocial behavior. PMID- 11931596 TI - Family discord, parental depression, and psychopathology in offspring: ten-year follow-up. AB - OBJECTIVE: To determine the independent effects of parental depression and family discord on psychopathology in offspring at high and low risk for major depression. METHOD: One hundred eighty-two offspring of depressed or nondepressed parents were followed over 10 years. In direct interviews, parents' and offspring's psychopathology was evaluated by raters blind to parents' clinical status. Five dimensions of family discord-poor marital adjustment, parent-child discord, low family cohesion, affectionless control, and parental divorce-were assessed. RESULTS: Offspring exposed to either parental depression or family discord had higher rates of psychopathology than their counterparts. High-risk offspring had few family discord measures associated with their psychopathology; in low-risk offspring, family discord was associated with all offspring diagnoses. Between the two risk factors, parental depression proved a more important predictor for offspring major depressive disorder (MDD) and anxiety disorder, whereas family discord was a more important predictor for substance use disorder. CONCLUSIONS: Parental depression is a strong and consistent risk factor for offspring MDD and anxiety disorder. Without parental depression, offspring have less exposure to family discord and lower rates of psychopathology. In the presence of family discord, rates of MDD, anxiety disorder and substance use disorder increased. When offspring matured into young adulthood, effects of parental depression and family discord persisted. PMID- 11931595 TI - Ethnic differences in comorbidity among substance-abusing adolescents referred to outpatient therapy. AB - OBJECTIVE: To examine differences in psychiatric comorbidity between African American and Hispanic substance-abusing adolescents referred for outpatient therapy. METHOD: Participants were 167 substance-abusing adolescents and their family members who completed an intake assessment. As part of the intake assessment, adolescents and parents were administered the Diagnostic Interview Schedule for Children-Predictive Scales to screen for the presence of nine psychiatric diagnoses representing both externalizing and internalizing disorders. RESULTS: Both African-American and Hispanic youths presented with high above-threshold symptom rates of co-occurring disorders. However, both adolescents and parents reported that Hispanic youths (78.3% and 83.9%, respectively) demonstrated greater rates of externalizing symptoms than African American youths (65.2% and 70.1%, respectively). African-American youths (40%) reported significantly more symptoms of agoraphobia than Hispanic youths (19.5%). CONCLUSIONS: The presence of high rates of co-occurring internalizing and externalizing problems provides evidence of the need for developing and implementing multifaceted interventions that address the complex emotional and behavioral needs of adolescent substance abusers. Among Hispanic youths in particular, treatments must address constellations of problem behaviors that appear to co-occur and likely represent the child's entrenchment in a deviant subculture. PMID- 11931597 TI - Depressive spectrum disorders in high-risk adolescents: episode duration and predictors of time to recovery. AB - OBJECTIVE: Durations of first episodes of major depression, dysthymia, adjustment disorders with depressed mood or with mixed emotional features, and minor depressions were examined in a community high-risk adolescent sample. Factors associated with longer recovery times were examined for each disorder. METHOD: Semistructured diagnostic interviews were conducted annually (6th through 12th grade) to assess children's current and lifetime history of psychopathology. Cox regressions were used to explore factors associated with longer time to recovery. RESULTS: Episode durations were consistent with prior community studies and generally shorter than those reported in clinical samples. A greater number of maternal depressive episodes predicted a more prolonged time to recovery from major depression, and greater adolescent impairment was associated with longer episode duration of both major and minor depression. CONCLUSIONS: Treatment and prevention efforts should be directed toward offspring of parents with recurrent mood disorders, particularly adolescents with minor as well as major depression. The authors also recommend that the median rather than the mean be used to evaluate the duration of depressive disorders. PMID- 11931598 TI - Anxiety and depression in children: a test of the positive-negative affect model. AB - OBJECTIVE: To examine whether the two-factor model of affect presented by Watson and colleagues is applicable to school children. METHOD: A child self-report scale of symptoms of depression and anxiety was administered to 624 school children in the third and sixth grades. Factor analyses were applied to extract factors of positive and negative affect. Reliabilities, correlations, and longitudinal analyses were performed to examine the developmental pathway of positive affect (PA) and negative affect (NA). RESULTS: In contrast to Watson's early findings, our results indicate that the PA and NA scales are significantly negatively correlated cross-sectionally as well as longitudinally. Moreover, our findings indicate that children with high NA and low PA in the third grade display a worse prognosis than children who have either high NA or low PA. CONCLUSIONS: A new affect model is needed for describing children's mood status. In addition, an important implication of these findings is that the occurrence of comorbid mood status in children confers a high risk for later emotional problems. PMID- 11931599 TI - Utah youth suicide study, phase I: government agency contact before death. AB - OBJECTIVE: To delineate the probability of contact between government agencies and youths who complete suicide, to investigate the nature of those contacts, and to identify new risk factors for suicide. METHOD: A descriptive epidemiological analysis of data from multiple Utah government agencies on consecutive youth suicides (n = 151) between August 1, 1996, and June 6, 1999, aged 13-21 years. Data were collected from four government agencies: Office of the Medical Examiner, Juvenile Justice, Department of Human Services, and the Department of Education. RESULTS: Utah descriptive data were similar to national statistics, with the majority of completers being male (89%) and firearms the most common method of suicide (58%). The data demonstrated an association between youth suicide and contact with Juvenile Justice. Sixty-three percent of youths who completed suicide in Utah had contact with Juvenile Justice, and there was a direct correlation between number of referrals and increased suicide risk. Suicide completers had multiple minor offenses over many years. A significant minority of school-age subjects could not be located within the school system. Few suicide completers had evidence of active psychiatric treatment. CONCLUSIONS: Juvenile Justice is identified as a novel site for the screening and identification of youths at risk for suicide. PMID- 11931600 TI - An intervention trial to improve adherence to community treatment by adolescents after a suicide attempt. AB - OBJECTIVE: To determine whether a problem-solving intervention would increase adherence to outpatient treatment for adolescents after a suicide attempt. METHOD: Sixty-three adolescents who had attempted suicide and were evaluated in an emergency department between 1997 and 2000 were randomly assigned to undergo standard disposition planning or a compliance enhancement intervention using a problem-solving format. At 3 months after the intervention, all evaluable adolescents, guardians, and outpatient therapists were contacted to determine adherence to outpatient treatment. RESULTS: At 3-month follow-up, the compliance enhancement group attended an average of 7.7 sessions compared with 6.4 sessions for the standard disposition group, but this difference was not statistically significant. However, after covarying barriers to receiving services in the community (such as being placed on a waiting list and insurance coverage difficulties), the compliance enhancement group attended significantly more treatment sessions than the standard disposition-planning group (mean = 8.4 versus 5.8 sessions). CONCLUSION: Interventions designed to improve treatment attendance must address not only individual and family factors but also service barriers encountered in the community that can impede access to services. PMID- 11931601 TI - Methylphenidate bioavailability in adults when an extended-release multiparticulate formulation is administered sprinkled on food or as an intact capsule. AB - OBJECTIVE: To determine the single-dose bioavailability of 20-mg Metadate CD (methylphenidate HCI, USP) Extended-Release Capsules sprinkled onto 1 level tablespoon (15 mL) of applesauce relative to an intact capsule under fasted conditions in healthy adults. METHOD: This was a single-center, open-label, single-dose, randomized, two-way crossover study with a 6-day washout period between doses, in healthy male and female subjects (N= 26), aged 21-40 years. Plasma concentration-time data for methylphenidate were used to calculate the pharmacokinetic parameters for each treatment. RESULTS: The pharmacokinetic profile for Metadate CD exhibited biphasic release characteristics with a sharp initial slope and a second rising portion. For Cmax (maximum observed concentration), AUC(0-infinity) (area under the plasma concentration curve from time 0 to infinity) and AUC(0-infinity) (area under the plasma concentration curve from time 0 to the last measurable time point), the geometric least squares mean ratios and 90% confidence intervals were within the 80% to 125% confidence interval for bioequivalence. Adverse events were similar to those reported for methylphenidate. CONCLUSIONS: The bioavailability of methylphenidate was not altered when Metadate CD capsules were administered by sprinkling their contents onto a small amount of applesauce. PMID- 11931602 TI - Children's trauma and adjustment reactions to violent and nonviolent war experiences. AB - OBJECTIVE: According to most studies, more than half of children exposed to war meet criteria for posttraumatic stress disorder (PTSD). Although we know that children are adversely affected by atrocities of war, we do not know which specific war events are associated with children's stress reactions. For example, it is unclear whether differences exist in response to violent versus nonviolent war-trauma experiences. This study examined the relationship of violent and nonviolent war experiences to children's trauma reactions and adjustment in a group of children from Bosnia. METHOD: During the 1994 siege in Sarajevo, 791 children aged 6 to 16 years participated in a study of trauma experience and response, in which assessment questionnaires (Impact of Event Scale, PTSD Reaction Index, Children's Depression Inventory, Child Behavior Checklist, and War Experience Questionnaire) were completed by children and their teachers. RESULTS: In this sample 41% had clinically significant PTSD symptoms. Children were adversely affected by exposure to both violent and nonviolent war-traumas. An additive effect of trauma exposure on trauma reactions was also found. However, many war experiences were not associated with children's adjustment and trauma reactions. CONCLUSIONS: Additive effects of violence and deprivations during war may overwhelm the coping skills of children and leave them vulnerable to externalizing and internalizing adjustment difficulties and symptoms of PTSD. PMID- 11931603 TI - Differential performance of the macarthur HBQ and DISC-IV in identifying DSM-IV internalizing psychopathology in young children. AB - OBJECTIVE: Three sites collaborated to evaluate the differential performance of the MacArthur Health and Behavior Questionnaire (HBQ) and the Diagnostic Interview Schedule for Children Version IV (DISC-IV) in identifying DSM-IV psychopathology in young children. METHOD: A sample of 120 four- to eight-year old nonreferred (community) (n = 67) and referred (clinical) (n = 53) children was examined. Mothers reported on their child's mental health symptoms using the HBQ (a dimensional measure with a clinical cutoff score) and the DISC-IV. Teachers independently reported on the child's symptoms and impairment in school using the teacher HBQ. Children self-reported on their symptoms using the companion Berkeley Puppet Interview. RESULTS: On the basis of its derived clinical cutoff scores, the HBQ identified significantly more children with clinical symptoms of DSM-IVinternalizing psychopathology than the DISC-IV in both referred and nonreferred groups. There was a high level of agreement between the two measures in the identification of externalizing psychopathology. Children identified as having internalizing psychopathology by the HBQ were also rated as highly symptomatic and impaired by teachers. CONCLUSION: The findings support the general validity of the parent HBQ for the assessment of young child psychopathology and the hypothesis that it captures more internalizing psychopathology than the DISC-IV in this population. PMID- 11931604 TI - Quantifying dimensions in autism: a factor-analytic study. AB - OBJECTIVE: The objective of this study was to determine whether the phenotypic variation in autism and the related pervasive developmental disorders (PDDs) is a unitary construct or whether it is composed of distinct dimensions of autistic symptoms and measures of level of functioning. METHOD: One hundred twenty-nine children with autism and other forms of PDD from two samples with different inclusion criteria were assessed with the Vineland Adaptive Behavior Scales to measure level of functioning and the Autism Diagnostic Interview to measure severity of autistic behaviors. A factor analysis with varimax rotation was performed on each sample, separately and combined. RESULTS: Two factors emerged; one representing autistic symptoms and another representing level of functioning. The factor structure was remarkably similar and robust to variations in ascertainment and inclusion criteria between the samples. The validity of the distinction was supported by differences between males and females on the symptom factor, but not on the level of functioning factor. IQ was modestly correlated with level of functioning, but not with symptoms. CONCLUSIONS: The phenotypic variation seen in autism/PDD is composed of at least two different dimensions of autistic symptoms and level of functioning. The implications of this dimensional heterogeneity for research, classification, and clinical practice are discussed. PMID- 11931605 TI - Premeditated mass shootings in schools: threat assessment. PMID- 11931606 TI - Selection of inferential statistics: an overview. AB - This column serves as an introduction to selection of appropriate statistical methods. In the next five columns we will discuss conceptually, and in more depth, these statistical methods. We will use clinical examples and discuss why the author(s) selected a particular statistical method and how the results of the statistical method were interpreted. PMID- 11931607 TI - Genetics of childhood disorders: XXXVII. News flash: a gene for expressive language. PMID- 11931608 TI - Inhibition of protein-protein association by small molecules: approaches and progress. PMID- 11931609 TI - Biological activity of the tryprostatins and their diastereomers on human carcinoma cell lines. AB - Tryprostatin A 1 and B 2 are indole alkaloid-based fungal products that act in the G2/M phase of the cell cycle. Tryprostatin A and B as well as their two enantiomers and four diastereomers have been synthesized via a common strategy. As a measure of cytotoxicity, these eight stereoisomers were assayed for their growth inhibitory properties in human breast, prostate, and lung cancer cell lines. The ability of the tryprostatins and the tryprostatin stereoisomers to induce topoisomerase II-mediated DNA relaxation or to inhibit tubulin polymerization was also examined. Although none of the stereoisomers were significantly active in topoisomerase II- or tubulin-based assays, ds2-try B 11 was found to exhibit a cytotoxicity profile more potent than etoposide 3 in the human cancer cell lines examined. In addition, ds2-try B 11 is comprised of an L tryptophan derivative coupled to a D-proline moiety, the latter stereochemistry of which may enhance the activity of 11 and potential analogues in vivo. PMID- 11931610 TI - Beta-C-mannosides as selectin inhibitors. AB - Potential E- and P-selectin inhibitors were synthesized to explore a hydrophobic area on the E-selectin surface and the PSGL-1 protein binding site on the P selectin surface that was recently defined by crystallography. Three series of mannose-based compounds (libraries A, B, and C) were synthesized using solution phase parallel synthesis. Biological evaluation of these compounds was done using two ELISA-based assays and transferred NOE (trNOE) experiments. Some of the compounds showed better activity than sLe(x) in the P-selectin assay. PMID- 11931611 TI - Synthesis, biological evaluation, and binding mode of novel 1-[2 (diarylmethoxy)ethyl]-2-methyl-5-nitroimidazoles targeted at the HIV-1 reverse transcriptase. AB - A novel series of 1-[2-(diarylmethoxy)ethyl]-2-methyl-5-nitroimidazole (DAMNI) analogues were synthesized and tested in cell-based assays and in enzyme assays against HIV-1 recombinant reverse transcriptase (RT). Preparation of the new derivatives was performed by reacting the appropriate benzhydrols or the corresponding bromides with 1-(2-hydroxyethyl)-2-methyl-5-nitroimidazole or the 3 hydroxypropyl homologue. Several compounds showed anti-HIV-1 activity in the submicromolar range. Structure-activity relationship studies suggested that meta substitution at one phenyl ring of the diarylmethane moiety strongly influences the antiviral activity. The 3,5-disubstitution at the same phenyl ring led to less potent derivatives. Molecular modeling and docking studies within the RT non nucleoside binding site confirmed that DAMNIs, similar to other NNRTIs such as TNK-651 and delavirdine (BHAP U90152), assume a butterfly-like conformation that appears to be halfway between that of classical NNRTIs, such as nevirapine, HEPT, TBZ, TIBO, and DABOs, and the conformation of BHAPs. In particular, the diphenylmethane moiety mimics the wings whereas the 1-(2-methyl-5 nitroimidazolyl)ethane portion resembles the BHAP 5-methanesulfonamidoindole-2 carbonylpiperazine portion. PMID- 11931613 TI - Development of new hydrogen-bond descriptors and their application to comparative molecular field analyses. AB - Knowledge-based descriptors extracted from composite crystal-field environments in crystal data have been developed for the description of interaction properties of small molecules. Using SuperStar seven diverse probe atoms have been selected to reflect the most important physicochemical properties. The general application of these descriptors in comparative molecular field analysis has been investigated using a dataset of thermolysin inhibitors, and a comparison to the GRID program has been performed. We especially focused on hydrogen-bond donor and acceptor properties by selecting a carbonyl and amino group as suitable probes. Their performance has been compared to that of the hydrogen-bond descriptors presently implemented in CoMSIA (comparative molecular similarity indices analysis). The newly developed descriptors produced significantly improved statistics for the correlation analyses if they are exclusively applied or, even better, applied in combination with other CoMSIA descriptors. Two methodologically different approaches have been tested to approximate the developed descriptors. Both reduce significantly the required computational efforts in particular for large data sets. The graphical interpretation of the field contributions of hydrogen-bonding properties elucidates additional features compared to those obtained from the original CoMSIA method. They are of valuable support for the design of improved inhibitors. PMID- 11931612 TI - GBR compounds and mepyramines as cocaine abuse therapeutics: chemometric studies on selectivity using grid independent descriptors (GRIND). AB - Cocaine is one of the most widely abused drugs in the industrial world. Substantial evidence has accumulated that the dopamine transporter (DAT) is a key target for cocaine regarding its reinforcing effects. This work describes the application of chemometric methods to a data set of 54 N(1)-benzhydryl-oxy-alkyl N(4)-phenyl-alk(en)yl-piperazines (GBR compounds) and chemically related mepyramines as putative candidates in cocaine abuse therapy. The aim of the study is to gain insight into the structural requirements that determine the affinity of the data set molecules to the DAT and the serotonin transporter (SERT) as well as their inhibitory potency on dopamine uptake. The compounds in the dataset are described using the recently developed GRID independent descriptors (GRIND), which allow one to obtain fast three-dimensional quantitative structure-activity relationship models without the need of aligning and superimposing the structures; the results are interpreted in a convenient pharmacophoric-like fashion. In the first part of the work, the selectivity of the database molecules for DAT binding vs dopamine reuptake inhibition is investigated. In the second part, the selectivity of the compounds for DAT binding vs SERT binding is studied. In both cases, significant models are obtained, which define the structural features responsible for the respective selectivity profiles. Moreover, the information has potential interest for the design of new derivatives with improved selectivity. PMID- 11931614 TI - Novel approach for linking genotype to phenotype in vitro by exploiting an extremely strong interaction between RNA and protein. AB - We recently isolated an aptamer that binds to the Tat protein of HIV-1 with extremely high affinity and specificity (Yamamoto, R.; et al. Genes Cells 2000, 5, 371.). In the present study, we exploited this strong binding to develop a novel coupling method that links genotype with phenotype. To strengthen the original RNA-protein interaction still further, we connected three units of the aptamer in tandem and three units of a peptide derived from Tat that interacted with the aptamer. The binding of the resultant RNA, which consisted of three units of the aptamer, to the resultant peptide, which consisted of three units of the peptide, was extremely strong. In fact, the RNA-protein interaction was one of the strongest ever reported, with an apparent K(d) below 16 pM. This strong interaction was attempted for the selection of functional proteins, namely, dihydrofolate reductase (DHFR) or streptavidin, which we chose as an example, and we succeeded in the expected selection, although to a limited extent, of the target protein. The noncovalent but strong interaction described above should be useful as a novel tool for the future selection of functional proteins from pools of random sequences of amino acids. PMID- 11931615 TI - Structure-based design, synthesis, and biological evaluation of irreversible human rhinovirus 3C protease inhibitors. 6. Structure-activity studies of orally bioavailable, 2-pyridone-containing peptidomimetics. AB - The structure-based design, chemical synthesis, and biological evaluation of various 2-pyridone-containing human rhinovirus (HRV) 3C protease (3CP) inhibitors are described. These compounds are comprised of a peptidomimetic binding determinant and a Michael acceptor moiety, which forms an irreversible covalent adduct with the active site cysteine residue of the 3C enzyme. The 2-pyridone containing inhibitors typically display improved 3CP inhibition properties relative to related peptide-derived molecules along with more favorable antiviral properties. The cocrystal structure of one pyridone-derived 3CP inhibitor complexed with HRV-2 3CP is also described along with certain ab initio conformation analyses. Optimization of the 2-pyridone-containing compounds is shown to provide several highly active 3CP inhibitors (k(obs)/[I] > 500,00 M(-1) s(-1)) that function as potent antirhinoviral agents (EC(50) = <0.05 microM) against multiple virus serotypes in cell culture. One 2-pyridone-containing 3CP inhibitor is shown to be bioavailable in the dog after oral dosing (F = 48%). PMID- 11931616 TI - Bioactive pseudopeptidic analogues and cyclostereoisomers of osteogenic growth peptide C-terminal pentapeptide, OGP(10-14). AB - The osteogenic growth peptide (OGP) is a key factor in the mechanism of the systemic osteogenic response to local bone marrow injury. When administered in vivo, OGP stimulates osteogenesis and hematopoiesis. The C-terminal pentapeptide OGP(10-14) is the minimal amino acid sequence that retains the full OGP-like activity. Apparently, it is also the physiologic active form of OGP. Residues Tyr(10), Phe(12), Gly(13), and Gly(14) of OGP are essential for the OGP(10-14) activity. The present study explored the functional role of the peptide bonds, carboxyl and amino terminal groups, and conformational freedom in OGP(10-14). Transformations replacing the peptide bonds with surrogates such as Psi(CH(2)NH), Psi(CONMe), and Psi(CH(2)CH(2)) demonstrated that amide bonds do not contribute significantly to OGP(10-14) bioactivity. End-to-end cyclization yielded the fully bioactive cyclic pentapeptide c(Tyr-Gly-Phe-Gly-Gly). The retroinverso analogue c(Gly-Gly-phe-Gly-tyr), a cyclostereoisomer of c(Tyr-Gly-Phe-Gly-Gly), is at least as potent as the parent cyclic pentapeptide. The unique structure-activity relations revealed in this study suggest that the spatial presentation of the Tyr and Phe side chains has a major role in the productive interaction of OGP(10-14) and its truncated and conformationally constrained analogues with their cognate cellular target. PMID- 11931617 TI - Structure-activity relationships in 1,4-benzodioxan-related compounds. 7. Selectivity of 4-phenylchroman analogues for alpha(1)-adrenoreceptor subtypes. AB - WB4101 (1)-related compounds 5-10 were synthesized, and their biological profile at alpha(1)-adrenoreceptor (AR) subtypes and 5-HT(1A) serotoninergic receptors was assessed by binding assays in Chinese hamster ovary and HeLa cell membranes expressing the human cloned receptors. Moreover, their receptor selectivity was further determined in functional experiments in isolated rat prostate (alpha(1A)), vas deferens (alpha(1A)), aorta (alpha(1D)), and spleen (alpha(1B)). In functional assays, compound 5 was the most potent at alpha(1D)-ARs with a reversed selectivity profile (alpha(1D) > alpha(1A) > alpha(1B)) relative to both prototype 1 and phendioxan (2) (alpha(1A) > alpha(1D) > alpha(1B)), whereas compound 8, bearing a carbonyl moiety at position 1, was the most potent at alpha(1A)-ARs with a selectivity profile similar to that of prototypes. The least potent of the series was the trans isomer 6, suggesting that optimum alpha(1)-AR blocking activity in this series is associated with a cis relationship between the 2-side chain and the 4-phenyl ring rather than a trans relationship as previously observed for the 2-side chain and the 3-phenyl ring in 2 and related compounds. Binding affinity results were not in complete agreement with the selectivity profiles deriving from functional experiments. Although a firm explanation was not available, neutral and negative antagonism and receptor dimerization were considered as two possibilities to account for the difference between binding and functional affinities. Finally, compound 5 was selected for a modeling study in comparison with 1, mephendioxan (3), and open phendioxan (4) to achieve information on the physicochemical interactions that account for its high affinity toward alpha(1d/D)-ARs. PMID- 11931618 TI - Design, synthesis, and structure-activity relationship of pyridyl imidazolidinones: a novel class of potent and selective human enterovirus 71 inhibitors. AB - When skeletons of Win compounds were used as templates, computer-assisted drug design led to the identification of a novel series of imidazolidinone derivatives with significant antiviral activity against enterovirus 71 (EV 71), the infection of which had resulted in about 80 fatalities during the 1998 epidemic outbreak in Taiwan. In addition to inhibiting all the genotypes (A, B, and C) of EV 71 in the submicromolar to low micromolar range, compounds 1 and 8 were extensively evaluated against a variety of viruses, showing potent activity against coxsackievirus A9 (IC(50) = 0.47-0.55 microM) and coxsackievirus A24 (IC(50) = 0.47-0.55 microM) as well as moderate activity against enterovirus 68 (IC(50) = 2.13 microM) and echovirus 9 (IC(50) = 2.6 microM). Our SAR studies revealed that imidazolidinone analogues with an aryl substituent at the para position of the phenoxyl ring, such as compounds 20, 21, 27, 57, 58, and 61, in general exhibited the highest activity against EV 71. Among them, compound 20 and its corresponding hydrochloride salt 57, in terms of potency and selectivity index, appear to be the most promising candidates in this series for further development of anti-EV 71 agents. Preliminary results of the study on the mode of action by a time course experiment suggest that test compounds 1 and 8 can effectively inhibit the virus replication at the early stages, referring to virus attachment or uncoating. This indicates that the surface protein may be the target for this type of compounds. PMID- 11931619 TI - Geometry-affinity relationships of the selective serotonin receptor ligand 9 (aminomethyl)-9,10-dihydroanthracene. AB - With the exception of its two aromatic rings and basic nitrogen atom, 9 (aminomethyl)-9,10-dihydroanthracene (AMDA; 1) is remarkably devoid of the pharmacophore features usually associated with high-affinity receptor ligands such as the heteroatom hydrogen bonding features of the endogenous ligand serotonin. AMDA does contain a phenylethylamine skeleton within a tricyclic ring system, and the presence of the second aromatic group is necessary for optimal receptor affinity. The structural requirements for the binding of AMDA at 5 HT(2A) receptors were investigated with respect to the geometric relationship between the two aromatic rings. It appears that the geometry of the AMDA parent is in the optimal range for fold angle between aromatic moieties. Evaluation of conformationally constrained derivatives of AMDA suggests that a chain extended trans, gauche form is most likely responsible for high affinity. PMID- 11931620 TI - Human somatostatin receptor specificity of backbone-cyclic analogues containing novel sulfur building units. AB - Somatostatin-14 (somatostatin) and its clinically available analogues octreotide, lanreotide, and vapreotide are potent inhibitors of growth hormone, insulin, and glucagon release. Recently, a novel backbone cyclic somatostatin analogue c(GABA Phe-Trp-(D)Trp-Lys-Thr-Phe-GlyC3-NH(2)) (analogue 1, PTR 3173) that possesses in vivo endocrine selectivity was described. This long-acting octapeptide exhibits high affinity to human recombinant somatostatin receptors (hsst) hsst2, hsst4, and hsst5. Its novel binding profile resulted in potent in vivo inhibition of growth hormone but not of insulin release. We report the synthesis, bioactivity, and structure-activity relationship studies of compounds related to 1. In these analogues, the lactam bridge of 1 was replaced by a backbone disulfide bridge. We present a novel approach for conformational constraint of peptides by utilizing sulfur-containing building units for on-resin backbone cyclization. These disulfide backbone cyclic analogues of 1 showed significant metabolic stability as tested in various enzyme mixtures. Receptor binding assays revealed different receptor selectivity profiles for these analogues in comparison to their prototype. It was found that analogues of 1, bearing a disulfide bridge, had increased selectivity to hsst2 and hsst5; however, they exhibited weaker affinity to hsst4 as compared to 1. These studies imply that ring chemistry, ring size, and ring position of the peptide template may affect the receptor binding selectivity. PMID- 11931621 TI - Gold(III) complexes with bipyridyl ligands: solution chemistry, cytotoxicity, and DNA binding properties. AB - Gold(III) compounds generally exhibit significant cytotoxic effects on cancer cell lines and are of potential interest as antitumor drugs. We report here on the solution chemistry, the cytotoxicity, and the DNA binding properties of two new bipyridyl gold(III) compounds: [Au(bipy)(OH)(2)][PF(6)] (1) and the organometallic compound [Au(bipy(c)-H)(OH)][PF(6)] (2) (bipy(c) = 6-(1,1 dimethylbenzyl)-2,2'-bipyridine). Both compounds are sufficiently soluble, and stable for hours, within a physiological buffer at 37 degrees C; [Au(bipy)(OH)(2)][PF(6)], at variance with [Au(bipy(c)-H)(OH)][PF(6)], is quickly and quantitatively reduced by ascorbate. Both compounds showed relevant cytotoxic effects toward the A2780S, A2780R, and SKOV3 tumor cell lines; lower effects were detected on the CCRF-CEM/S and CCRF-CEM/R lines. In most cases the mechanisms of resistance to CDDP are only marginally effective against these gold(III) complexes. The interactions of [Au(bipy)(OH)(2)][PF(6)] and [Au(bipy(c) H)(OH)][PF(6)] with calf thymus DNA were investigated in vitro by various techniques to establish whether DNA represents a primary target for these compounds. Addition of saturating amounts of DNA did not affect appreciably the visible spectra of these gold(III) complexes. Some slight modifications of the CD spectra of calf thymus DNA and of the DNA melting parameters were observed; in any case, ultrafiltration experiments showed that binding of these gold(III) complexes to DNA is weak and reversible. The mechanistic implications of these findings are discussed. PMID- 11931622 TI - Synthesis of photoreactive phosphatidic acid analogues displaying activatory properties on cyclic AMP-phosphodiesterases. Photoaffinity labeling of an isoform of phosphodiesterase. AB - We have previously shown that phosphatidic acid (PA) is a specific activator of some isoforms of type 4 cyclic nucleotide phosphodiesterases (PDE 4) and that accumulation of endogenous PA can, in this way, influence the cAMP signaling pathway in different cell types. Enzyme activation depends on direct binding of the effector to specific sites carried by the enzyme. To identify the binding domain, photoactivatable phosphatidic acid analogues 1-azidoPA (12) and 2-azidoPA (7 and 15), potentially suitable for covalent labeling of PDE4, have been synthesized. The ability of phospholipases A(2) and D to hydrolyze unnatural phospholipids has been considered in this paper. The effect of 1-azidoPA (12) and 2-azidoPA (7 and 15) on the activity of a recombinant PA-sensitive isoform PDE4D3 was evaluated. The three compounds were able to activate the enzyme with different efficiencies. A tritiated analogue of 15 was synthesized and used in PDE4D3 labeling experiments, which showed that this PA analogue was specifically and covalently linked to the enzyme after UV irradiation. Photoactivatable analogues thus appear as suitable tools for the characterization of PA binding sites. PMID- 11931623 TI - Synthesis and biological characterization of L-N(6)-(1-iminoethyl)lysine 5 tetrazole-amide, a prodrug of a selective iNOS inhibitor. AB - The 5-tetrazole amide of L-N(6)-(1-iminoethyl)lysine (L-NIL), L-N(6)-(1 iminoethyl)lysine 5-tetrazole amide (1), has been prepared and evaluated. In contrast to L-NIL, 1 is a stable, nonhygroscopic, crystalline solid. Unlike L NIL, 1 has minimal inhibitory activity in vitro on human inducible nitric oxide synthase (iNOS). However, it is rapidly converted in vivo to L-NIL and produces dose-dependent inhibition of iNOS in acute and chronic models of inflammation in the rodent with efficacy comparable to L-NIL. In addition, both 1 and L-NIL exhibit significant and comparable in vivo selectivity for the inhibition of iNOS vs endothelial NOS. Doses approximately 80-fold greater than those that inhibited inflammation do not elevate systemic blood pressure. In summary, both the physical properties and the pharmacological profile of 1 make it an ideal molecule for preclinical and clinical studies on the role of selective iNOS inhibitors in mediating inflammatory disease processes. PMID- 11931624 TI - Synthesis and in vitro antitumor activity of new deaza analogues of the nonpolyglutamatable antifolate N(alpha)-(4-amino-4-deoxypteroyl)-N(delta) hemiphthaloyl-L-ornithine (PT523). AB - Details are disclosed for the synthesis of N(alpha)-[4-[2-(2,4-diaminoquinazolin 6-yl)ethyl]benzoyl]-N(delta)-hemiphthaloyl-L-ornithine (2) and N(alpha)-[4-[5 (2,4-diaminoteridin-6-yl)pent-1-yn-4-yl]benzoyl]-N(delta)-hemiphthaloyl-L ornithine (6) as analogues of N(alpha)-(4-amino-4-deoxypteroyl)-N(delta) hemiphthaloyl-L-ornithine (1, PT523), a nonpolyglutamatable antifolate currently in advanced preclinical development. In a 72 h growth inhibition assay against cultures of CCRF-CEM human leukemic lymphoblasts, the IC(50) of 2 and 6 was 0.69 +/- 0.044 nM and 1.3 +/- 0.35 nM, respectively, as compared with previously reported values 4.4 +/- 0.10 nM for aminopterin (AMT) and 1.5 +/- 0.39 nM for PT523. In a spectrophotometric assay of dihydrofolate reductase (DHFR) inhibition using dihydrofolate and NADPH as the cosubstrates, the previously unreported compounds 2 and the mixed 10R and 10S diastereomers of 6 had K(i) values of 0.21 +/- 0.05 pM and 0.60 +/- 0.02 pM, respectively, as compared with previously reported values of 3.70 +/- 0.35 pM for AMT and 0.33 +/- 0.04 pM for PT523. Thus, while they were comparable to 1 and several of its previously studied analogues in their ability to bind to DHFR and inhibit the growth of CCRF-CEM cells, 2 and the mixed diastereomers of 6 were several times more active than AMT despite the fact that they cannot form gamma-polyglutamylated metabolites of the type formed in cells from AMT and other classical antifolates with a glutamate side chain. PMID- 11931625 TI - Potent, orally active heterocycle-based combretastatin A-4 analogues: synthesis, structure-activity relationship, pharmacokinetics, and in vivo antitumor activity evaluation. AB - The synthesis and structure-activity relationship study of a series of compounds with heterocycles in place of the cis double bond in combretastatin A-4 (CA-4) are described. Substituted tosylmethyl isocyanides were found to be the key intermediates in construction of the heterocycles. Cytotoxicities of the heterocycle-based CA-4 analogues were evaluated against NCI-H460 and HCT-15 cancer cell lines. 3-Amino-4-methoxyphenyl and N-methyl-indol-5-yl were the best replacements for the 3-hydroxy-4-methoxyphenyl in CA-4. 4,5-Disubstituted imidazole was found to be the best for the replacement of the cis double bond in CA-4. Medicinal chemistry efforts led to the discovery of compounds 24h and 25f that were found to be 32 and 82% bioavailable, respectively, in rat. Evaluation of 24h and 25f against murine M5076 reticulum sarcoma in mice revealed that both compounds were orally efficacious with an increase in life span of 38.5 and 40.5%, respectively. PMID- 11931626 TI - A common mechanism underlying promiscuous inhibitors from virtual and high throughput screening. AB - High-throughput and virtual screening are widely used to discover novel leads for drug design. On examination, many screening hits appear non-drug-like: they act noncompetitively, show little relationship between structure and activity, and have poor selectivity. Attempts to develop these peculiar molecules into viable leads are often futile, and much time can be wasted on the characterization of these "phony" hits. Despite their common occurrence, the mechanism of action of these promiscuous molecules remains unknown. To investigate this problem, 45 diverse screening hits were studied. Fifteen of these were previously reported as inhibitors of various receptors, including beta-lactamase, malarial protease, dihydrofolate reductase, HIV Tar RNA, thymidylate synthase, kinesin, insulin receptor, tyrosine kinases, farnesyltransferase, gyrase, prions, triosephosphate isomerase, nitric oxide synthase, phosphoinositide 3-kinase, and integrase; 30 were from an in-house screening library of a major pharmaceutical company. In addition to their original targets, 35 of these 45 compounds were shown to inhibit several unrelated model enzymes. These 35 screening hits included compounds, such as fullerenes, dyes, and quercetin, that have repeatedly shown activity against diverse targets. When tested against the model enzymes, the compounds showed time-dependent but reversible inhibition that was dramatically attenuated by albumin, guanidinium, or urea. Surprisingly, increasing the concentration of the model enzymes 10-fold largely eliminated inhibition, despite a 1000-fold excess of inhibitor; a well-behaved competitive inhibitor did not show this behavior. One model to explain these observations was that the active form of the promiscuous inhibitors was an aggregate of many individual molecules. To test this hypothesis, light scattering and electron microscopy experiments were performed. The nonspecific inhibitors were observed to form particles of 30 400 nm diameter by both techniques. In control experiments, a well-behaved competitive inhibitor and an inactive dye-like molecule were not observed to form aggregates. Consistent with the hypothesis that the aggregates are the inhibitory species, the particle size and IC(50) values of the promiscuous inhibitors varied monotonically with ionic strength; a competitive inhibitor was unaffected by changes in ionic strength. Unexpectedly, aggregate formation appears to explain the activity of many nonspecific inhibitors and may account for the activity of many promiscuous screening hits. Molecules acting via this mechanism may be widespread in drug discovery screening databases. Recognition of these compounds may improve screening results in many areas of pharmaceutical interest. PMID- 11931627 TI - Conceptually new low-calcemic oxime analogues of the hormone 1 alpha,25 dihydroxyvitamin D(3): synthesis and biological testing. AB - New chemical entities 16-ene-25-ketone 2b and the corresponding oxime 3b and oxime ether 4b, analogues of natural calcitriol (1), were rationally designed and synthesized on a milligram scale. Chemical introduction of the oxime ether functionality in analogue 4b was successful via direct oximation of an intact vitamin D conjugated triene system. Even though all three analogues are at least as antiproliferative in vitro as calcitriol (1) even at physiologically relevant low nanomolar concentrations, only side chain ketone 2b is more transcriptionally potent than calcitriol (1). Although oxime O-methyl ether 4b lacks the traditional side chain hydrogen bond-donating OH group of the natural hormone and lacks also the oxime-NOH group of analogue 3b, surprisingly, oxime ether 4b retains 20% of the transcriptional potency of natural calcitriol (1). In terms of in vivo toxicity (hypercalcemia), ketone 2b is strongly calcemic in rats, whereas oxime 3b and oxime ether 4b are considerably less calcemic (i.e., safer) than calcitriol (1). PMID- 11931628 TI - A comparative molecular field analysis (COMFA) of the structural determinants of heat-stable enterotoxins mediating activation of guanylyl cyclase C. AB - The heat-stable enterotoxin binds to and activates guanylyl cyclase C (GC-C), regulating fluid and electrolyte secretion in intestinal epithelial cells. A COMFA model was developed to predict the primary interactions between GC-C agonists and their receptor. This model predicts that the amide backbone of Cys(5)-Cys(6)-Glu(7)-Leu(8), the beta carbon atoms of Cys(5)-Cys(6), and the side chains of Pro(12), Ala(13), and Ala(15) comprise the primary interactions of GC-C agonists with the receptor surface. PMID- 11931630 TI - Integrin-linked kinase phosphorylates the myosin phosphatase target subunit at the inhibitory site in platelet cytoskeleton. AB - The myosin phosphatase (MP) composed of the catalytic subunit of type 1 protein phosphatase and myosin phosphatase target subunit isoform 1 (MYPT1) was identified as the major serine/threonine phosphatase component in the platelet cytoskeleton fraction. MYPT1 was phosphorylated by cytoskeletal kinase(s), but the identity of the kinase(s) and the effect of phosphorylation were not established. Incubation of platelet-cytoskeletal fraction with MgATP or MgATP[S] (magnesium adenosine 5'-[gamma-thio]triphosphate) caused a decrease in the 20 kDa light-chain of smooth-muscle myosin (MLC20) phosphatase and phosphorylase phosphatase activities. MYPT1 contains a phosphorylation site, Thr-695, involved in the inhibition of MP in a RhoA/Rho kinase-dependent manner. The cytoskeletal kinase(s) phosphorylated Thr-695 of glutathione S-transferase (GST)-MYPT1, as determined with an antibody specific for phosphorylated Thr-695. The level of Rho kinase was low in the cytoskeletal fraction and was detected primarily in the membrane and cytosolic fractions. The phosphorylation of Thr-695 by the cytoskeletal kinase(s) was not affected by Rho kinase inhibitor, Y-27632, suggesting that kinase(s) other than Rho kinase were involved. In-gel kinase assay identified a kinase at 54-59 kDa that phosphorylated the C-terminal fragment of MYPT1 (GST-MYPT1(667-1004)). Western blots detected both zipper interacting protein kinase (ZIPK) and integrin-linked kinase (ILK) at 54-59 kDa in the cytoskeleton and membrane fractions. Cytoskeletal ZIPK and ILK were separated and partially purified by chromatography on SP-Sepharose and on MonoQ. ZIPK preferentially phosphorylated MLC20 and had low activity on MYPT1. ILK phosphorylated both MLC20 and MYPT1 and phosphorylation of MYPT1 occured on Thr 695. The above results raise the potential for regulation of MP activity in platelet cytoskeleton by ILK and suggest an alternative to the Rho-linked pathway. PMID- 11931631 TI - Functional studies on human Pex7p: subcellular localization and interaction with proteins containing a peroxisome-targeting signal type 2 and other peroxins. AB - Pex7p is a WD40-containing protein involved in peroxisomal import of proteins containing an N-terminal peroxisome-targeting signal (PTS2). The interaction of human recombinant Pex7p expressed in different hosts/systems with its PTS2 ligand and other peroxins was analysed using various experimental approaches. Specific binding of human Pex7p to PTS2 could be demonstrated only when Pex7p was formed in vitro by a coupled transcription/translation system or synthesized in vivo in Chinese hamster ovary K1 cells transfected with a construct coding for a Pex7p green fluorescent protein (GFP) fusion protein. Apparently, no cofactors are required and only monomeric Pex7p binds to PTS2. The interaction is reduced upon cysteine alkylation and is impaired upon truncation of the N-terminus of Pex7p. Interaction of Pex7p with other peroxins could not be demonstrated in bacterial or yeast two-hybrid screens, or in pull-down binding assays. The GFP fusion proteins, tagged at either the N- or C-terminus, were able to restore PTS2 import in rhizomelic chondrodysplasia punctata fibroblasts, and Pex7p-GFP was located both in the lumen of peroxisomes and in the cytosol. PMID- 11931632 TI - Control of domain swapping in bovine odorant-binding protein. AB - As revealed by the X-ray structure, bovine odorant-binding protein (OBPb) is a domain swapped dimer [Tegoni, Ramoni, Bignetti, Spinelli and Cambillau (1996) Nat. Struct. Biol. 3, 863-867; Bianchet, Bains, Petosi, Pevsner, Snyder, Monaco and Amzel (1996) Nat. Struct. Biol. 3, 934-939]. This contrasts with all known mammalian OBPs, which are monomers, and in particular with porcine OBP (OBPp), sharing 42.3% identity with OBPb. By the mechanism of domain swapping, monomers are proposed to evolve into dimers and oligomers, as observed in human prion. Comparison of bovine and porcine OBP sequences pointed at OBPp glycine 121, in the hinge linking the beta-barrel to the alpha-helix. The absence of this residue in OBPb might explain why the normal lipocalin beta-turn is not formed. In order to decipher the domain swapping determinants we have produced a mutant of OBPb in which a glycine residue was inserted after position 121, and a mutant of OBPp in which glycine 121 was deleted. The latter mutation did not result in dimerization, while OBPb-121Gly+ became monomeric, suggesting that domain swapping was reversed. Careful structural analysis revealed that besides the presence of a glycine in the hinge, the dimer interface formed by the C-termini and by the presence of the lipocalins conserved disulphide bridge may also control domain swapping. PMID- 11931633 TI - Effect of inositol 1,4,5-trisphosphate receptor stimulation on mitochondrial [Ca2+] and secretion in chromaffin cells. AB - Ca(2+) uptake by mitochondria is a potentially important buffering system able to control cytosolic [Ca(2+)]. In chromaffin cells, we have shown previously that stimulation of either Ca(2+) entry or Ca(2+) release via ryanodine receptors triggers large increases in mitochondrial [Ca(2+)] ([Ca(2+)](M)) approaching the millimolar range, whose blockade dramatically enhances catecholamine secretion [Montero, Alonso, Carnicero, Cuchillo-Ibanez, Albillos, Garcia, Carcia-Sancho and Alvarez (2000) Nat. Cell Biol. 2, 57-61]. In the present study, we have studied the effect of stimulation of inositol 1,4,5-trisphosphate (InsP(3)) receptors using histamine. We find that histamine produces a heterogeneous increase in [Ca(2+)](M), reaching peak levels at approx. 1 microM in 70% of the mitochondrial space to several hundred micromolar in 2-3% of mitochondria. Intermediate levels were found in the rest of the mitochondrial space. Single-cell imaging experiments with aequorin showed that the heterogeneity had both an intercellular and a subcellular origin. Those mitochondria responding to histamine with increases in [Ca(2+)](M) much greater than 1 microM (30%) were the same as those that also responded with large increases in [Ca(2+)](M) following stimulation with either high-K(+) medium or caffeine. Blocking mitochondrial Ca(2+) uptake with protonophores or mitochondrial inhibitors also enhanced catecholamine secretion induced by histamine. These results suggest that some InsP(3) receptors tightly co-localize with ryanodine receptors and voltage-dependent Ca(2+) channels in defined subplasmalemmal functional units designed to control secretion induced by different stimuli. PMID- 11931634 TI - Sheep have an unusual variant of the brain-specific metallothionein, metallothionein-III. AB - Sheep metallothionein-III (MT-III) cDNA was isolated from a brain cDNA library and characterized. In contrast with MT-III from other species, sheep MT-III cDNA is predicted to encode a protein with significantly different metal-binding properties, owing to the loss of three of its cysteine residues. RT-PCR from other sheep confirmed that this aberrant structure is ubiquitous in this species. MT-III was successfully isolated from sheep brain, demonstrating that the cDNA does give rise to a protein product of the predicted structure. Sheep MT-III is similar to other mammalian MT-IIIs in that it retains the Cys-Pro-Cys-Pro motif which is thought to encode growth-inhibitory activity, and we show that it is likewise able to inhibit neuron survival in vitro. This is the first naturally occurring variant of MT-III (or any other major mammalian MT gene) which gives rise to a protein product. These findings are discussed in light of proposed roles of MT in the mammalian brain. PMID- 11931635 TI - Loop variants of the serpin thyroxine-binding globulin: implications for hormone release upon limited proteolysis. AB - Thyroxine-binding globulin (TBG) and corticosteroid-binding globulin are unique among non-inhibitory members of the superfamily of serine-proteinase inhibitors (serpins) in undergoing a dramatic increase in stability [stressed-to-relaxed (S- >R) transition] after proteolytic cleavage within their exposed reactive-site loop (RSL) equivalent. This structural rearrangement involves the insertion of the cleaved loop as a new strand into the beta-sheet A and is accompanied by a decrease in hormone binding. To define the mechanism that leads to disruption of hormone binding of TBG after proteolytic cleavage, the effect of partial loop deletions and replacements by the alpha(1)-proteinase inhibitor homologues of TBG were evaluated. Unexpectedly, deletion of the loop's C-terminus, thought to be important for thyroxine binding, improved the binding affinity over that of normal TBG. Proteolytic cleavage of this variant revealed an intact S-->R transition and reduced its binding activity to that of cleaved TBG. In contrast, a chimaera with C-terminal loop extension mimicked the decreased binding affinity of cleaved TBG and had a thermal stability intermediate between that of native and cleaved serpins. This variant was still susceptible to loop cleavage and underwent an S-->R transition, yet without changing its binding affinity. Our data exclude a direct involvement of loop residues in thyroxine binding of native TBG. Limited insertion of the RSL into beta-sheet A appears to trigger hormone release after proteolytic cleavage. In support of this concept, residues within the hinge region of the TBG loop are phylogenetically highly conserved, suggestive of their physiological role as a functional switch in vivo. PMID- 11931637 TI - Characterization and expression of human bifunctional 3'-phosphoadenosine 5' phosphosulphate synthase isoforms. AB - Sulphonation, a fundamental process essential for normal growth and development, requires the sulphonate donor molecule 3'-phosphoadenosine 5'-phosphosulphate (PAPS), which is produced from ATP and inorganic sulphate by the bifunctional enzyme PAPS synthase. In humans, two genes encode isoenzymes that are 77% identical at the amino acid level, and alternative splicing creates two subtypes of PAPS synthase 2. The question as to whether distinctions in amino acid composition are reflected in differences in activity has been examined. The specific activity of the PAPS synthase 2 subtypes is 10- to 15-fold higher than that for PAPS synthase 1. The greater catalytic efficiency of the PAPS synthase 2 subtypes is demonstrated further by the 3- to 6-fold higher k(cat)/K(m) ratios for ATP and inorganic sulphate as compared with the ratios for PAPS synthase 1. In humans, PAPS synthase 1 is expressed ubiquitously, and is the dominant isoform in most tissues, whereas expression of the PAPS synthase 2 subtypes is variable and tissue-specific. It is noteworthy that, similar to other human tissues, PAPS synthase 1 also appears to be the dominant isoform expressed in cartilage. The latter finding initially created a conundrum, since there is a specific human dwarfing disorder that is known to be caused by a mutation in the PAPS synthase 2 gene. This apparent enigma would seem to be resolved by examination of cartilage from guinea-pigs as an animal model. Similar to humans, cartilage from mature animals predominantly expresses PAPS synthase 1. In contrast, expression of PAPS synthase 1 is relatively low in the cartilage of immature guinea-pigs, including the growth plate of long bones, whereas PAPS synthase 2 is the highly expressed isoenzyme. PMID- 11931638 TI - The Saccharomyces cerevisiae 14-3-3 protein Bmh2 is required for regulation of the phosphorylation status of Fin1, a novel intermediate filament protein. AB - In order to identify proteins that interact with Bmh2, a yeast member of the 14-3 3 protein family, we performed a two-hybrid screening using LexA-Bmh2 as bait. We identified Fin1, a novel intermediate filament protein, as the protein that showed the highest degree of interaction. We also identified components of the vesicular transport machinery such as Gic2 and Msb3, proteins involved in transcriptional regulation such as Mbf1, Gcr2 and Reg2, and a variety of other different proteins (Ppt1, Lre1, Rps0A and Ylr177w). We studied the interaction between Bmh2 and Fin1 in more detail and found that Bmh2 only interacted with phosphorylated forms of Fin1. In addition, we showed that Glc7, the catalytic subunit of the protein phosphatase 1 complex, was also able to interact with Fin1. PMID- 11931639 TI - Vps4-A (vacuolar protein sorting 4-A) is a binding partner for a novel Rho family GTPase, Rnd2. AB - Rho family GTPases are implicated in a variety of biological activities, including endocytic vesicle trafficking. Rnd2 is a new member of Rho family GTPases, but its biological functions are not known. In the present study, we have performed a yeast two-hybrid screening using Rnd2 as bait and revealed that Rnd2 binds specifically to Vps4-A (where Vsp4-A is vacuolar protein sorting 4-A), a member of the AAA ATPase family and a central regulator for early endosome trafficking. This interaction was determined by the yeast two-hybrid system, in vitro binding and co-immunoprecipitation studies. Vps4-A associated with both guanosine 5'-[beta-thio]triphosphate-bound active and guanosine 5'-[beta thio]diphosphate-bound inactive forms of Rnd2. An ATPase-defective Vps4-A mutant, Vps4-A(E228Q), expressed in HeLa cells was accumulated in the early endosomes. When Rnd2 was co-expressed with Vps4-A(E228Q), Rnd2 was recruited to the Vps4-A bound early endosomes. These results suggest that Rnd2 is involved in the regulation of endosomal trafficking via direct binding to Vps4-A. PMID- 11931640 TI - Ppm1, a novel polyprenol monophosphomannose synthase from Mycobacterium tuberculosis. AB - Dolichol monophosphomannose (DPM) is an ever-present donor of mannose (Man) in various eukaryotic glycosylation processes. Intriguingly, the related polyprenol monophosphomannose (PPM) is involved in the biosynthesis of lipomannan and lipoarabinomanan, key bacterial factors termed modulins that are found in mycobacteria. Based on similarities to known DPM synthases, we have identified and characterized the PPM synthase of Mycobacterium tuberculosis, now termed Mt Ppm1. In the present study, we demonstrate that Mt-Ppm1 possesses an unusual two domain architecture, by which the second domain is sufficient for PPM synthesis. However, when overexpressed separately in mycobacteria, domain 1 of Mt-Ppm1 appears to increase the synthesis of PPM. Interestingly, other mycobacteria such as M. smegmatis, M. avium and M. leprae produce two distinct proteins, which are similar to the two domains found in Mt-Ppm1. Using an in vitro assay, we also demonstrate that Mt-Ppm1 transfers Man from GDP-Man to a structurally diverse range of lipid monophosphate acceptors. The identification of the PPM synthase as a key enzyme in lipoarabinomannan biosynthesis now provides an attractive candidate for gene disruption to generate mutants for subsequent immunological studies. PPM synthase can also be exploited as a target for specific inhibitors of M. tuberculosis. PMID- 11931641 TI - Phosphorylation-dependent interaction of the synaptic vesicle proteins cysteine string protein and synaptotagmin I. AB - The secretory vesicle cysteine string proteins (CSPs) are members of the DnaJ family of chaperones, and function at late stages of Ca2+-regulated exocytosis by an unknown mechanism. To determine novel binding partners of CSPs, we employed a pull-down strategy from purified rat brain membrane or cytosolic proteins using recombinant hexahistidine-tagged (His(6)-)CSP. Western blotting of the CSP binding proteins identified synaptotagmin I to be a putative binding partner. Furthermore, pull-down assays using cAMP-dependent protein kinase (PKA) phosphorylated CSP recovered significantly less synaptotagmin. Complexes containing CSP and synaptotagmin were immunoprecipitated from rat brain membranes, further suggesting that these proteins interact in vivo. Binding assays in vitro using recombinant proteins confirmed a direct interaction between the two proteins and demonstrated that the PKA-phosphorylated form of CSP binds synaptotagmin with approximately an order of magnitude lower affinity than the non-phosphorylated form. Genetic studies have implicated each of these proteins in the Ca2+-dependency of exocytosis and, since CSP does not bind Ca2+, this novel interaction might explain the Ca2+-dependent actions of CSP. PMID- 11931642 TI - Mechanism in the reaction of cytochrome c oxidase with organic hydroperoxides: an ESR spin-trapping investigation. AB - Organic hydroperoxides are of great utility in probing the reaction mechanism and the toxicological consequences of lipid peroxidation. In the present study, ESR spin-trapping was employed to investigate the peroxidation of mitochondrial cytochrome c oxidase (CcO) with t-butyl hydroperoxide (t-BuOOH) and cumene hydroperoxide (CumOOH). The spin trap 5,5-dimethyl-1-pyrroline N-oxide (DMPO) was used to detect the radical species formed from the reaction of CcO with t-BuOOH. The presence of t-BuOOH-derived alkoxyl radical (t-BuO*) as the primary radical indicates reductive scission of the O-O bond by CcO. The ESR signal of DMPO/*Ot Bu can be partially abolished by cyanide, implying that the reductive cleavage involved the haem a(3)Cu(B) binuclear site of CcO. A nitroso spin trap, 2-methyl 2-nitrosopropane (MNP), was used to detect and identify radical species from the reaction of CcO with CumOOH. In addition to the t-BuOOH-derived methyl, hydroxylmethyl and tertiary carbon-centred radicals, a protein-derived radical was detected. The intensity of the ESR signal from the protein radical increased with the CumOOH concentration at low CumOOH/CcO ratios, with maximal intensity at a ratio of 100 mol of CumOOH/mol of CcO. The immobilized protein radical adduct of MNP was stable and persistent after dialysis; it was also resistant to proteolytic digestion, suggesting that it was formed in the transmembrane region, a region that is not accessible to proteases. Its signal was greatly enhanced when CcO cysteine residues were chemically modified by N-ethylmaleimide, when the tryptophan residues in CcO were oxidized by N-bromosuccimide, and when tyrosine residues on the surface of CcO were iodinated, showing that a radical equilibrium was established among the cysteine, tryptophan and tyrosine residues of the protein-centred radical. Pre-treatment of CcO with cyanide prevented detectable MNP adduct formation, confirming that the haem a(3)-Cu(B) binuclear centre was the initial reaction site. When the CcO was pre-treated with 10 mM (100 equivalents) of CumOOH, the enzyme activity decreased by more than 20%. This inhibition was persistent after dialysis, suggesting that the detected protein centred radical was, in part, involved in the irreversible inactivation by CumOOH. Visible spectroscopic analysis revealed that the haem a of CcO was not affected during the reaction. However, the addition of pyridine to the reaction mixture under alkaline conditions resulted in the destruction of the haem centre of CcO, suggesting that its protein matrix rather than its haem a is the target of oxidative damage by the organic hydroperoxide. PMID- 11931643 TI - XpsG, the major pseudopilin in Xanthomonas campestris pv. campestris, forms a pilus-like structure between cytoplasmic and outer membranes. AB - GspG, -H, -I, -J and -K proteins are members of the pseudopilin family. They are the components required for the type II secretion pathway, which translocates proteins across the outer membrane of Gram-negative bacteria to the extracellular milieu. They were predicted to form a pilus-like structure, and this has been shown for PulG of Klebsiella oxytoca by using electron microscopy. In the present study, we performed biochemical analyses of the XpsG protein of Xanthomonas campestris pv. campestris and observed that it is a pillar-like structure spanning the cytoplasmic and outer membranes. Subcellular fractionation revealed a soluble form (SF) of XpsG, in addition to the membrane form. Chromatographic analysis of SF XpsG in the absence of a detergent indicated that it is part of a large complex (>440 kDa). In vitro studies indicated that XpsG is prone to aggregate in the absence of a detergent. We isolated and characterized a non functional mutant defective in forming the large complex. It did not interfere with the function of wild-type XpsG and was not detectable in the SF. Moreover, unlike wild-type XpsG, which was distributed in both the cytoplasmic and outer membranes, it appeared only in the cytoplasmic membrane. When wild-type XpsG was co-expressed with His6-tagged XpsH but not with untagged XpsH, SF XpsG bound to nickel and co-eluted with XpsH. This result suggests the presence of other pseudopilin components in the XpsG-containing large-sized molecules. PMID- 11931646 TI - Hepatocyte growth factor activates phosphoinositide 3-kinase C2 beta in renal brush-border plasma membranes. AB - Upon stimulation of renal cortical slices with hepatocyte growth factor (HGF), inositol lipid metabolism was studied in basal-lateral plasma membranes (BLM) and brush-border plasma membranes (BBM). Whereas in BLM rapid increases in 1,2 diacylglycerol, PtdIns(3,4,5)P(3) and PtdIns(3,4)P(2) were observed, suggesting that in BLM HGF activates both phospholipase C (PLC) and phosphoinositide 3 kinase (PI3K), in BBM only HGF-induced transient accumulation of PtdIns3P was seen, which was temporarily delayed from signalling events in BLM and could be blocked by the PtdIns-specific-PLC inhibitor ET-18-OCH(3) and the calpain inhibitor calpeptin, suggesting that 3-kinase activation in BBM lies downstream of PLC activation in BLM and is a calpain-mediated event. Moreover, the increase in immunoprecipitable PI3K-C2 beta activity, which is sensitive to wortmannin (10 nM) and shows strong preference for PtdIns over PtdIns4P as a substrate, was observed only in BBM upon stimulation of renal cortical slices with HGF and could be mimicked by the Ca(2+) ionophore A23187 and blocked by the cell-penetrant Ca(2+) chelator BAPTA-AM [1,2-bis-(2-aminophenoxy)ethane-N,N,N',N'-tetra-acetic acid tetrakis(acetoxymethyl ester)]. On Western blots PI3K-C2 beta revealed a single immunoreactive band of 180 kDa in BLM and BBM, while after stimulation with HGF a gel shift of 18 kDa was noticed only in BBM, suggesting that the observed enzyme activation is achieved by proteolysis. When BBM were subjected to short-term (15 min) exposure to mu-calpain, a similar gel shift together with an increase in PI3K-C2 beta activity was observed, when compared with the BBM harvested after HGF stimulation. The above-mentioned gel shift and increase in PI3K-C2 beta activity could be prevented by the calpain inhibitor calpeptin. The data presented in this report show that in renal cells there is a spatial separation of the inositol lipid signalling system between BLM and BBM, and that HGF causes activation of PLC and PI3K primarily in BLM, which leads to calpain mediated activation of PI3K-C2 beta in BBM with a concomitant increase in PtdIns3P. PMID- 11931647 TI - The role of syndecan-2 in regulation of actin-cytoskeletal organization of Lewis lung carcinoma-derived metastatic clones. AB - Syndecans, a family of transmembrane heparan sulphate proteoglycans, contribute to various biological processes, including adhesion, motility, proliferation, differentiation and morphogenesis. We document here the involvement of syndecan-2 acting alone or co-operatively with integrin alpha5beta1, for regulation of actin cytoskeletal organization on cell adhesion to fibronectin, using fibronectin recombinant polypeptides containing the ligands for either or both of these receptors as substrata. Lewis lung carcinoma-derived low-metastatic P29 cells binding to the substrata by both receptors formed actin stress fibres, whereas those binding by syndecan-2 or integrin alpha5beta1 alone formed filopodia or cortex actin. In contrast, higher metastatic LM66-H11 cells formed cortex actin even on substrata containing both ligands. Northern-blot and flow-cytometric analyses revealed that syndecan-2 expression in LM66-H11 cells was significantly lower (1/4.5 in mRNA and 1/8 in cell-surface expression) than in P29 cells, whereas expression levels of integrin alpha5beta1 and other syndecans were similar in both cell types. These results suggest that the failure of LM66-H11 to form stress fibres is due to a lower expression of syndecan-2 than that due to a threshold for its function. This was confirmed by the finding that overexpression of syndecan-2 by transfection of its cDNA into LM66-H11 cells caused the formation of stress fibres on the fibronectin substratum. These in vitro cellular responses of the two clones might reflect their in vivo situation in primary tumours in which P29 cells with a stroma-inducing capacity were immediately surrounded by fibronectin-rich matrix formed by the induced stromal cells, whereas LM66-H11 cells without such capacity were not surrounded by a similar matrix. PMID- 11931648 TI - Mitogen-activated protein kinase-dependent and -independent routes control shedding of transmembrane growth factors through multiple secretases. AB - Solubilization of a number of membrane proteins occurs by the action of cell surface proteases, termed secretases. Recently, the activity of these secretases has been reported to be controlled by the extracellular signal-regulated kinases 1 and 2 (ERK1/ERK2) and the p38 mitogen-activated protein kinase (MAPK) routes. In the present paper, we show that shedding of membrane-anchored growth factors (MAGFs) may also occur through MAPK-independent routes. In Chinese-hamster ovary cells, cleavage induced by protein kinase C (PKC) stimulation was largely insensitive to inhibitors of the ERK1/ERK2 and p38 routes. Other reagents such as sorbitol or UV light stimulated MAGF cleavage independent of PKC. The action of sorbitol on cleavage was only partially prevented by the combined action of inhibitors of the p38 and ERK1/ERK2 routes, indicating that sorbitol can also stimulate shedding by MAPK-dependent and -independent routes. Studies in cells devoid of activity of the secretase tumour necrosis factor-alpha-converting enzyme (TACE) indicated that this protease had an essential role in PKC- and ERK1/ERK2-mediated shedding. However, secretases other than TACE may also cleave MAGFs since sorbitol could still induce shedding in these cells. These observations suggest that cleavage of MAGFs is a complex process in which multiple secretases, activated through different MAPK-dependent and -independent routes, are involved. PMID- 11931649 TI - Efficient identification of photolabelled amino acid residues by combining immunoaffinity purification with MS: revealing the semotiadil-binding site and its relevance to binding sites for myristates in domain III of human serum albumin. AB - To identify photoaffinity-labelled amino acid residue(s), we devised an effective method utilizing immunoaffinity purification of photolabelled fragments, followed by matrix-assisted laser-desorption ionization-time of flight (MALDI-TOF) MS and nanoelectrospray ionization tandem MS (nano-ESI-MS/MS) analysis. Human serum albumin (HSA) was photolabelled with an azidophenyl derivative of semotiadil, FNAK [(+)-(R)-3,4-dihydro-2-[5-methoxy-2-[3-[N-methyl-N-[2-(3-azidophenoxy) ethyl]amino]propoxyl]phenyl]-4-methyl-2H-1,4-benzothiazin-3-(4H)-one], since HSA is a major binding protein for semotiadil in serum. After lysyl endopeptidase digestion, photolabelled HSA fragments were adsorbed selectively on to Sepharose beads on which an anti-semotiadil antibody was immobilized, and fractions were eluted quantitatively by 50% acetonitrile/10 mM HCl. MALDI-TOF MS analysis of the eluted fraction showed that it contained two photolabelled fragments of m/z 2557.54 (major) and 1322.44 (minor), corresponding to Lys-414-Lys-432 and Ala-539 Lys-545, respectively. Further nano-ESI-MS/MS analysis revealed that Lys-414 was the photolabelled amino acid residue in fragment 414-432 and Lys-541 was a likely candidate in fragment 539-545. Based on the photolabelling results, we constructed a three-dimensional model of the FNAK-HSA complex, revealing that FNAK resides in a pocket that overlaps considerably with myristate (Myr)-binding sites, Myr-3 and -4, by comparison with crystallographic data of HSA-Myr complexes described in Curry, Mandelkow, Brick and Franks (1998) Nat. Struct. Biol. 5, 827-835. Moreover, addition of Myr increased photo-incorporation into Lys-414, whereas incorporation into Lys-541 decreased under conditions of [Myr]/[HSA]<1. Further addition of Myr, however, uniformly decreased photo incorporation into both Lys residues. These results indicate that FNAK labelling can also be used to monitor Myr binding in domain III. An interpretation for the concomitant local conformational change of HSA is provided. PMID- 11931644 TI - Structure-function of the multifunctional Ca2+/calmodulin-dependent protein kinase II. AB - Ca2+/calmodulin (CaM)-dependent protein kinase (CaMKII) is a ubiquitous mediator of Ca2+-linked signalling that phosphorylates a wide range of substrates to co ordinate and regulate Ca2+-mediated alterations in cellular function. The transmission of information by the kinase from extracellular stimuli and the intracellular Ca2+ rise is not passive. Rather, its multimeric structure and autoregulation enable this enzyme to participate actively in the sensitivity, timing and location of its action. CaMKII can: (i) be activated in a Ca2+-spike frequency-dependent manner; (ii) become independent of its initial Ca2+/CaM activators; and (iii) undergo a 'molecular switch-like' behaviour, which is crucial for certain forms of learning and memory. CaMKII is derived from a family of four homologous but distinct genes, with over 30 alternatively spliced isoforms described at present. These isoforms possess diverse developmental and anatomical expression patterns, as well as subcellular localization. Six independent catalytic/autoregulatory domains are connected by a narrow stalk-like appendage to each hexameric ring within the dodecameric structure. Ca2+/CaM binding activates the enzyme by disinhibiting the autoregulatory domain; this process initiates an intra-holoenzyme autophosphorylation reaction that induces complex changes in the enzyme's sensitivity to Ca2+/CaM, including the generation of Ca2+/CaM-independent (autonomous) activity and marked increase in affinity for CaM. The role of CaMKII in Ca2+ signal transduction is shaped by its autoregulation, isoenzymic type and subcellular localization. The molecular determinants and mechanisms producing these processes are discussed as they relate to the structure-function of this multifunctional protein kinase. PMID- 11931650 TI - Actopaxin is phosphorylated during mitosis and is a substrate for cyclin B1/cdc2 kinase. AB - Prior to cell division, normal adherent cells adopt a round morphology that is associated with a loss of actin stress fibres and disassembly of focal adhesions. In this study, we investigate the mitotic phosphorylation of the recently described paxillin and actin-binding focal-adhesion protein actopaxin [Nikolopoulos and Turner (2000) J. Cell Biol. 151, 1435-1448]. Actopaxin is comprised of an N-terminus containing six putative cdc2 phosphorylation sites and a C-terminus consisting of tandem calponin homology domains. Here we show that the N-terminus of actopaxin is phosphorylated by cyclin B1/cdc2 kinase in vitro and that this region of actopaxin precipitates cdc2 kinase activity from mitotic lysates. Actopaxin exhibits reduced electrophoretic mobility during mitosis that is dependent on phosphorylation within the first two consensus cdc2 phosphorylation sites. Finally, as cells progress from mitosis to G(1) there is an adhesion-independent dephosphorylation of actopaxin, suggesting that actopaxin dephosphorylation precedes cell spreading and the reformation of focal adhesions. Taken together, these results suggest a role for cyclin B1/cdc2-dependent phosphorylation of actopaxin in regulating actin cytoskeleton reorganization during cell division. PMID- 11931651 TI - Molecular dissection of membrane-transport proteins: mass spectrometry and sequence determination of the galactose-H+ symport protein, GalP, of Escherichia coli and quantitative assay of the incorporation of [ring-2-13C]histidine and (15)NH(3). AB - The molecular mass of the galactose-H(+) symport protein GalP, as its histidine tagged derivative GalP(His)(6), has been determined by electrospray MS (ESI-MS) with an error of <0.02%. One methionine residue, predicted to be present from the DNA sequence, was deduced to be absent. This is a significant advance on the estimation of the molecular masses of membrane-transport proteins by SDS/PAGE, where there is a consistent under-estimation of the true molecular mass due to anomalous electrophoretic migration. Addition of a size-exclusion chromatography step after Ni(2+)-nitrilotriacetate affinity purification was essential to obtain GalP(His)(6) suitable for ESI-MS. Controlled trypsin, trypsin+chymotrypsin and CNBr digestion of the protein yielded peptide fragments suitable for ESI-MS and tandem MS analysis, and accurate mass determination of the derived fragments resulted in identification of 82% of the GalP(His)(6) protein. Tandem MS analysis of selected peptides then afforded 49% of the actual amino acid sequence of the protein; the absence of the N-terminal methionine was confirmed. Matrix-assisted laser-desorption ionization MS allowed identification of one peptide that was not detected by ESI-MS. All the protein/peptide mass and sequence determinations were in accord with the predictions of amino acid sequence deduced from the DNA sequence of the galP gene. [ring-2-(13)C]Histidine was incorporated into GalP(His)(6) in vivo, and ESI-MS analysis enabled the measurement of a high (80%) and specific incorporation of label into the histidine residues in the protein. MS could also be used to confirm the labelling of the protein by (15)NH(3) (93% enrichment) and [(19)F]tryptophan (83% enrichment). Such MS measurements will serve in the future analysis of the structures of membrane-transport proteins by NMR, and of their topology by indirect techniques. PMID- 11931652 TI - Promoter characterization of the novel human matrix metalloproteinase-26 gene: regulation by the T-cell factor-4 implies specific expression of the gene in cancer cells of epithelial origin. AB - A novel matrix metalloproteinase-26 (MMP-26) is known to be specifically expressed in epithelial carcinomas. To facilitate studies of MMP-26 transcriptional regulation, we have cloned and characterized a 1 kb 5'-flanking region of the human MMP-26 gene. Altogether, our findings indicate that the MMP 26 promoter has distinctive structural and functional features among MMP genes. An unusual polyadenylation site proximal to the transcription-factor-binding sites protects transcription of the MMP-26 gene from the upstream promoters and represents a part of the stringent transcriptional regulation of the gene. The MMP-26 gene has a consensus TATA-box and one transcriptional start site located 60 and 35 nucleotides upstream of the translational start site, respectively. The MMP-26 promoter was able to drive luciferase expression in human A549 lung carcinoma, HT1080 fibrosarcoma and HEK293 embryonic kidney cells. The basal transcription efficiency of the MMP-26 promoter is relatively low, thereby explaining the minute expression of the gene in most cells and tissues. When compared with other MMP genes, the MMP-26 promoter contains binding sites for a few transcription factors. Sequential deletion and mutation analysis, and electrophoretic mobility-shift assay have identified the T-cell factor-4 (Tcf-4) motif and the activator protein-1 site as the major regulatory elements of the MMP-26 promoter. Since previous studies have established that the Tcf-4 transcription factor is subjected exclusively to regulation through the beta catenin/E(epithelial)-cadherin pathway, this implies the specific expression of MMP-26 in cancer cells of epithelial origin. PMID- 11931653 TI - Transcriptional regulation of human 3'-phosphoadenosine 5'-phosphosulphate synthase 2. AB - Sulphonation is a fundamental process that is essential for normal growth and development as well as maintenance of the internal milieu. The universal sulphonate donor molecule essential for all sulphoconjugation reactions is adenosine 3'-phosphate 5'-phosphosulphate (PAPS), which is produced from ATP and inorganic sulphate by the action of bifunctional PAPS synthase. There are two isozymes encoded by genes located on chromosome 4 (PAPS synthase 1) and chromosome 10 (PAPS synthase 2). The promoter for PAPS synthase 2 contains neither a TATAAA nor a CCAAT box, although a consensus initiator motif is present. Three human cell lines were used to examine promoter activity after transfection with various lengths of the 5'-flanking region of the PAPS synthase 2 gene fused to a reporter gene. Proximal promoter activity was located between bp -84 and bp -124 upstream of the purported transcription start site. This region contains two GC/GT boxes that are essential for full promoter activity, as indicated by deletion analysis and supported further by mutagenesis. A nuclear extract of SW13 cells, which highly express PAPS synthase 2, contained proteins that bound to probes possessing promoter-specific GC/GT boxes. Furthermore, the presence of specificity protein (Sp) 1, Sp2 and Sp3 proteins in the nuclear extract was confirmed by supershift analysis. Co-transfection experiments using SL2 cells yielded additional support for the involvement of Sp1 in transcriptional regulation of the PAPS synthase 2 gene; the involvement of Sp2 and/or Sp3 remains to be clarified further. PMID- 11931654 TI - beta1 and beta2 integrins activate different signalling pathways in monocytes. AB - Integrin-mediated signals play an important but poorly understood role in regulating many leucocyte functions. In monocytes and macrophages, integrins of the beta2 subfamily are involved in cell-cell interactions that are important for migration of the cells through the endothelium and also for phagocytosis. On the other hand, in the same cells, beta1 integrin-mediated adhesion to extracellular matrix proteins results in a strong induction of immediate early genes that are important in inflammation. To investigate the signalling pathways from these two types of integrin in monocytic cells, THP-1 cells were selectively stimulated via beta1 or beta2 integrins by cross-linking each type of receptor with specific monoclonal antibodies or their natural ligands. The involvement of extracellular signal-regulated kinase (ERK), Syk and phosphoinositide 3-kinase (PI-3K) was then analysed. Nuclear factor kappaB (NF-kappaB) activation was also detected in THP-1 cells transiently transfected with an NF-kappaB-driven luciferase reporter gene. We found that binding of both types of integrin to their natural ligands activated ERK in a Syk- and PI-3K-dependent manner. Yet, cross-linking of integrins by anti-beta1 antibodies caused activation of ERK while that by anti beta2 antibodies did not. Also both types of integrin activated NF-kappaB. However, PI-3K was required for beta1 integrin-, but not beta2 integrin-, mediated NF-kappaB activation. In addition, inhibition of PI-3K with wortmannin and LY294002 blocked beta1 integrin-mediated NF-kappaB activation, but did not affect that mediated by beta2 integrin. These data suggest that distinct integrins activate different signalling pathways in monocytic cells. PMID- 11931655 TI - Identification of protein-derived tyrosyl radical in the reaction of cytochrome c and hydrogen peroxide: characterization by ESR spin-trapping, HPLC and MS. AB - The reaction of cytochrome c and H(2)O(2) is known to form a protein-centred radical that can be detected with the spin trap 2-methyl-2-nitrosopropane (MNP). To characterize the MNP/tyrosyl adduct structure that had previously been determined incorrectly [Barr, Gunther, Deterding, Tomer and Mason (1996) J. Biol. Chem. 271, 15498-15503], we eliminated unreasonable structure models by ESR studies with a series of (13)C-labelled tyrosines, and photochemically synthesized an authentic MNP/tyrosyl adduct that has its trapping site on the C-3 position of the tyrosine phenyl ring. The observation of the identical ESR spectra for this radical adduct from the UV irradiation of 3-iodo-tyrosine and the adduct from the cytochrome c reaction demonstrated that the radical trapping site of MNP/tyrosyl is located on the equivalent C-3/C-5 positions instead of the C-1 position, as was proposed by Barr et al. In an on-line HPLC/ESR system, an identical retention time (17.7 min) was observed for the ESR-active HPLC peak of the MNP/tyrosyl adduct from the following three reactions: (i) the tyrosine oxidation via horseradish peroxidase/H(2)O(2); (ii) UV irradiation of 3-iodo tyrosine and (iii) the reaction of cytochrome c with H(2)O(2). This result demonstrated that the radical adducts of all three reactions are most probably the same. The mass spectrometric analysis of the HPLC fractions from reactions (i) and (ii) showed an ion at m/z 267 attributed to the MNP/tyrosyl adduct. We conclude that the cytochrome c-derived tyrosyl radical was trapped by MNP, leading to a persistent radical adduct at the C-3/C-5 positions of the tyrosine phenyl ring. PMID- 11931656 TI - Regulation and recruitment of phosphatidylinositol 4-kinase on immature secretory granules is independent of ADP-ribosylation factor 1. AB - Heterotrimeric G-proteins, as well as small GTPases of the Rho and ADP ribosylation factor (ARF) family, are implicated in the regulation of lipid kinases, including PtdIns 4-kinases and PtdIns(4)P 5-kinases. Here, we describe a PtdIns 4-kinase activity on immature secretory granules (ISGs), regulated secretory organelles formed from the trans-Golgi network (TGN), and investigate the regulation of PtdIns4P levels on these membranes. Over 50% of the PtdIns 4 kinase activity on ISGs is inhibited by both a low concentration of adenosine and the monoclonal antibody 4C5G, a specific inhibitor of the type II PtdIns 4 kinase. Treatment of ISGs with mastoparan 7 (M7) stimulates the type II PtdIns 4 kinase via pertussis-toxin-sensitive G(i)/G(0) proteins, which, in contrast with previous results obtained with chromaffin granules [Gasman, Chasserot-Golaz, Hubert, Aunis and Bader (1998) J. Biol. Chem. 273, 16913-16920], does not require Rho A, B or C. M7 treatment also leads to an inhibition in the recruitment of ARF to ISG membranes: this inhibition is not dependent on G(i)/G(0) activation, and is not linked to the stimulation of PtdIns 4-kinase observed with M7. PtdIns 4 kinase activity on ISGs is not regulated by myristoylated ARF1-GTP, in contrast with results obtained with Golgi membranes [Godi, Pertile, Meyers, Marra, Di Tullio, Iurisci, Luini, Corda and De Matteis (1999) Nat. Cell Biol. 1, 280-287; Jones, Morris, Morgan, Kondo, Irvine and Cockcroft (2000) J. Biol. Chem. 275, 13962-13170], whereas ARF1-GTP does regulate the production of PtdIns(4,5)P(2). Our results suggest that the regulation of PtdIns 4-kinase on the ISGs differs in comparison with that on the TGN, and might be related to a specific requirement of ISG maturation. PMID- 11931657 TI - Cloning and expression of two novel pig liver and kidney fatty acid hydroxylases [cytochrome P450 (CYP)4A24 and CYP4A25]. AB - A new member of the cytochrome P450 (CYP) 4A subfamily (CYP4A21) was recently cloned by PCR from pig liver [Lundell, Hansson, and Wikvall (2001) J. Biol. Chem. 276, 9606-9612]. This enzyme does not catalyse omega- or (omega-1)-hydroxylation of lauric acid, the model substrate for CYP4A enzymes. Instead, CYP4A21 participates in bile acid biosynthesis in the pig. Extensive studies, primarily conducted to verify the aberrant amino acids found in CYP4A21 within a normally conserved CYP4A motif, revealed that besides CYP4A21 two additional sequences were co-amplified by PCR. These two sequences (designated CYP4A24 and CYP4A25), generated from both pig liver and kidney, were characterized by restriction enzyme analysis and were subsequently cloned. The deduced amino acid sequences of CYP4A24 and CYP4A25 share extensive sequence identity (97%). Both enzymes, expressed in yeast cells, exhibit omega-and (omega-1)-hydroxylase activities towards lauric acid and palmitic acid. The positions of the variable regions between CYP4A24 and CYP4A25, which are confined to beta-sheets 1 and 4, indicate a possible difference in substrate specificity or regioselectivity. The porcine CYP4A21, CYP4A24 and CYP4A25 enzymes, with an overall identity of 94%, have probably evolved from a common ancestral gene, perhaps in conjunction with species-specific habits. PMID- 11931658 TI - Modulation of Mycobacterium tuberculosis DnaA protein-adenine-nucleotide interactions by acidic phospholipids. AB - The biochemical aspects of the initiation of DNA replication in Mycobacterium tuberculosis are unknown. To understand this process, we overproduced, purified and characterized the recombinant M. tuberculosis DnaA protein. The M. tuberculosis DnaA protein binds the origin of replication (oriC), ATP and ADP, and exhibited weak ATPase activity. ADP, after hydrolysis of ATP, remained strongly associated with DnaA and the exchange of ATP for bound ADP was weak. Vesicles prepared from acidic phospholipids, such as phosphatidylinositol, cardiolipin and phosphatidylglycerol, promoted dissociation of both ADP and ATP, whereas the neutral phospholipid phosphatidylethanolamine did not. The phospholipid-mediated dissociation of ATP was decreased in the presence of the M. tuberculosis oriC, whereas dissociation of ADP was stimulated in the presence of oriC. Acidic phospholipids in micelles, however, were not efficient in dissociating bound nucleotides from DnaA. Together, these results suggest that both polar head groups and membrane bilayer structure play an important role in M. tuberculosis DnaA-adenine-nucleotide interactions. We suggest that initiation of M. tuberculosis oriC involves intimate interactions between DnaA, adenine nucleotides and membrane phospholipids, and the latter helps to ensure that only the ATP form of the DnaA protein interacts continuously with oriC. PMID- 11931659 TI - Folate synthesis in higher-plant mitochondria: coupling between the dihydropterin pyrophosphokinase and the dihydropteroate synthase activities. AB - The plant enzyme 6-hydroxymethyl-7,8-dihydropterin pyrophosphokinase/7,8 dihydropteroate synthase (HPPK/DHPS) is a mitochondrial bifunctional protein involved in tetrahydrofolate synthesis. The first domain (HPPK) catalyses the pyrophosphorylation of 6-hydroxymethyl-7,8-dihydropterin (dihydropterin) by ATP, leading to 6-hydroxymethyl-7,8-dihydropterin pyrophosphate (dihydropterinPP(i)) and AMP. The second domain (DHPS) catalyses the next step, i.e. the condensation of p-aminobenzoic acid (p-ABA) with dihydropterinPP(i) to give 7,8 dihydropteroate (dihydropteroate) and PP(i). In the present article we studied the coupling between these two reactions. Kinetic data obtained for the HPPK domain are consistent with an ordered Bi Bi mechanism where ATP binds first and dihydropterinPP(i) is released last, as proposed previously for the monofunctional Escherichia coli enzyme. In the absence of p-ABA, AMP and dihydropterinPP(i) accumulate and negatively regulate the reaction. In the presence of p-ABA, the rates of AMP and dihydropteroate synthesis are similar, indicating a good coupling between the two reactions. DihydropterinPP(i), an intermediate of the two reactions, never accumulates in this situation. The high specific activity of DHPS relative to HPPK, rather than a preferential channelling of dihydropterinPP(i) between the two catalytic sites, could explain these kinetic data. The maximal velocity of the DHPS domain is limited by the availability of dihydropterinPP(i). It is strongly feedback-inhibited by dihydropteroate and also dihydrofolate and tetrahydrofolate monoglutamate, two intermediates synthesized downstream in the folate biosynthetic pathway. Thus the HPPK domain of this bifunctional protein is the limiting factor of the overall reaction, but the DHPS domain is a potential key regulatory point of the whole folate biosynthetic pathway. PMID- 11931661 TI - The reassembling process of the nonameric Mycobacterium tuberculosis small heat shock protein Hsp16.3 occurs via a stepwise mechanism. AB - Conditions are reported under which the reassembled intermediates of the heat shock protein Hsp16.3 after being denatured in 8 M urea were detected by mainly using urea-gradient PAGE (with modifications) and urea-denaturing pore-gradient PAGE. Hsp16.3 is the small heat-shock protein from Mycobacterium tuberculosis, which exists as a specific nonamer and was proposed to form a trimer-of-trimers structure. The refolding and reassembling of this protein was achieved rapidly by dilution or dialysis, suggesting an effectively spontaneous recovery of quaternary structure. Data presented in this report demonstrate that the in vitro reassembling process of Hsp16.3 protein occurs through a spontaneous and effective stepwise mechanism. Modified urea-gradient PAGE may provide a general method for studying the reassembling processes of other oligomeric proteins. PMID- 11931660 TI - Copper supplementation restores cytochrome c oxidase activity in cultured cells from patients with SCO2 mutations. AB - Human SCO2 is a nuclear-encoded Cu-binding protein, presumed to be responsible for the insertion of Cu into the mitochondrial cytochrome c oxidase (COX) holoenzyme. Mutations in SCO2 are associated with cardioencephalomyopathy and COX deficiency. Studies in yeast and bacteria have shown that Cu supplementation can restore COX activity in cells harbouring mutations in genes involving Cu transport. Therefore we investigated whether Cu supplementation could restore COX activity in cultured cells from patients with SCO2 mutations. Our data demonstrate that the COX deficiency observed in fibroblasts, myoblasts and myotubes from patients with SCO2 mutations can be restored to almost normal levels by the addition of CuCl(2) to the growth medium. PMID- 11931662 TI - Substrate-binding recognition and specificity of trehalose phosphorylase from Schizophyllum commune examined in steady-state kinetic studies with deoxy and deoxyfluoro substrate analogues and inhibitors. AB - Trehalose phosphorylase is a component of the alpha-D-glucopyranosyl alpha-D glucopyranoside (alpha,alpha-trehalose)-degrading enzyme system in fungi and it catalyses glucosyl transfer from alpha,alpha-trehalose to phosphate with net retention of the anomeric configuration. The enzyme active site has no detectable affinity for alpha,alpha-trehalose in the absence of bound phosphate and catalysis occurs from the ternary complex. To examine the role of non-covalent enzyme-substrate interactions for trehalose phosphorylase recognition, we used the purified enzyme from Schizophyllum commune and tested a series of incompetent structural analogues of the natural substrates and products as inhibitors of the enzyme. Equilibrium-binding constants (K(i)) for deoxy- and deoxyfluoro derivatives of D-glucose show that loss of interactions with the 3-, 4- or 6-OH, but not the reactive 1- and the 2-OH, results in considerably (> or =100-fold) weaker affinity for sugar-binding subsite +1, revealing the requirement for hydrogen bonding with hydroxyls, away from the site of chemical transformation to position precisely the D-glucose-leaving group/nucleophile for catalysis. The high specificity of trehalose phosphorylase for the sugar aglycon during binding and conversion of O-glycosides is in contrast with the observed alpha-retaining phosphorolysis of alpha-D-glucose-1-fluoride (alpha-D-Glc-1-F) since the productive bonding capability of the fluoride-leaving group with subsite +1 is minimal. The specificity constant (19 M(-1).s(-1)) and catalytic-centre activity (0.1 s(-1)) for the reaction with alpha-D-Glc-1-F are 0.10- and 0.008-fold the corresponding kinetic parameters for the enzymic reaction with alpha,alpha trehalose. The non-selective-inhibition profile for a series of inactive alpha-D glycopyranosyl phosphates shows that the driving force for the binary-complex formation lies mainly in interactions of the enzyme with the phosphate group and suggests that hydrogen bonding with hydroxyl groups at the catalytic site (subsite -1) contributes to catalysis by providing stabilization, which is specific to the transition state. Vanadate, a tight-binding phosphate mimic, inhibits the phosphorolysis of alpha-D-Glc-1-F by forming a ternary complex whose apparent dissociation constant of 120 microM is approx. 160-fold greater than the dissociation constant of the same inhibitor complex with alpha,alpha-trehalose. PMID- 11931663 TI - Thermodynamics of the ligandin function of human class Alpha glutathione transferase A1-1: energetics of organic anion ligand binding. AB - In addition to their catalytic functions, cytosolic glutathioneS-transferases (GSTs) are a major reserve of high-capacity binding proteins for a large variety of physiological and exogenous non-substrate compounds. This ligandin function has implicated GSTs in numerous ligand-uptake, -transport and -storage processes. The binding of non-substrate ligands to GSTs can inhibit catalysis. In the present study, the energetics of the binding of the non-substrate ligand 8 anilino-1-naphthalene sulphonate (ANS) to wild-type human class Alpha GST with two type-1 subunits (hGSTA1-1) and its DeltaPhe-222 deletion mutant were studied by isothermal titration calorimetry. The stoichiometry of binding to both proteins is one ANS molecule per GST subunit with a greater affinity for the wild type (K(d)=65 microM) than for the DeltaPhe-222 mutant (K(d)=105 microM). ANS binding to the wild-type protein is enthalpically driven and it is characterized by a large negative heat-capacity change, DeltaC(p). The negative DeltaC(p) value for ANS binding indicates a specific interface with a significant hydrophobic component in the protein-ligand complex. The negatively charged sulphonate group of the anionic ligand is apparently not a major determinant of its binding. Phe 222 contributes to the binding affinity for ANS and the hydrophobicity of the binding site. PMID- 11931664 TI - Expression in mammalian cell cultures reveals interdependent, but distinct, functions for Star and Rhomboid proteins in the processing of the Drosophila transforming-growth-factor-alpha homologue Spitz. AB - We report here distinct interdependent functions for two proteins, Star and Rhomboid, that are key determinants of the epidermal-growth-factor (EGF)-receptor signalling pathway in Drosophila. When we expressed the Drosophila EGF-receptor ligand Spitz in mammalian cells, the protein failed to traffic to the plasma membrane, as assessed by either cell-surface protein biotinylation or immunocytochemical staining. However, when we co-expressed Star with Spitz, trafficking of Spitz to the cell surface could be demonstrated. Only when we co expressed Spitz, Star and Rhomboid could the release of soluble Spitz protein into the medium be shown. Taken together, our results indicate that Star is required for the intracellular trafficking of Spitz, and that Rhomboid is essential for the release of soluble Spitz protein from cells. PMID- 11931665 TI - Phenylalanine-24 in the N-terminal region of ammodytoxins is important for both enzymic activity and presynaptic toxicity. AB - Ammodytoxins (Atxs) are group II phospholipases A(2) (PLA(2)s) with presynaptic toxicity from venom of the snake Vipera ammodytes ammodytes. The molecular basis of their neurotoxicity, and that of similar PLA(2) toxins, is still to be explained. To address this problem, a surface-exposed aromatic residue, Phe(24), in the N-terminal region of the most potent Atx, AtxA, was replaced by other aromatic (tyrosine, tryptophan), hydrophobic (alanine) and polar uncharged (serine, asparagine) residues. The mutants were produced in the bacterial expression system, refolded in vitro and purified to homogeneity. All but the Trp(24) mutant, whose activity was similar to that of the wild type, showed a considerable decrease (40-80%) in enzymic activity on a micellar phosphatidylcholine substrate. This result indicates an important role for the aromatic side chains of phenylalanine or tryptophan, but not tyrosine, in PLA(2) activity, very likely at a stage of interfacial adsorption of the enzyme to zwitterionic aggregated substrates. The substitutions of Phe(24) also significantly decreased toxicity in mice, with the most prominent decrease, of 130-fold, observed in the case of the Asn(24) mutant. The results with the mutants show that there is no correlation between enzymic activity, lethality and binding affinity for three AtxA neuronal receptors (R180, R25 and calmodulin). Our results suggest a critical involvement of Phe(24) in the neurotoxicity of AtxA, apparently at a stage which does not involve the interaction with the known Atx-binding neuronal proteins and catalytic activity. PMID- 11931666 TI - Activation of rhodopsin kinase. AB - The present study confirms our original assertion that peptides corresponding to the C-terminal sequence of rhodopsin are phosphorylated by rhodopsin kinase (RK), but only in the presence of photo-activated rhodopsin [Rho*, which is functionally equivalent to metarhodopsin II (Meta II)]. Under optimized conditions, the extent of peptide phosphorylation reached up to 60% that of Rho*. Rho* phosphorylation began to plateau within 15 min of the initiation of photolysis, whereas the peptide phosphorylation continued linearly for >60 min. This lack of co-ordination in the phosphorylation of the physiological (Rho*) and synthetic (peptide) substrates necessitated the present study, which showed that RK is activated for peptide phosphorylation not only by Meta II but also by Meta III, as well as by the phosphorylated derivatives of these species, but not by opsin. These results led to the conclusion that all the derivatives of opsin, which contain the Schiff base linkage with the all-trans-retinylidene moiety, retain the ability to activate RK. PMID- 11931667 TI - Intracellular trafficking of glycosylphosphatidylinositol (GPI)-anchored proteins and free GPIs in Leishmania mexicana. AB - Free glycosylphosphatidylinositols (GPIs) are an important class of membrane lipids in many pathogenic protozoa. In this study, we have investigated the subcellular distribution and intracellular trafficking of an abundant class of free GPIs [termed glycosylinositolphospholipids (GIPLs)] in Leishmania mexicana promastigotes. The intracellular transport of the GIPLs and the major GPI anchored glycoprotein gp63 was measured by following the incorporation of these molecules into sphingolipid-rich, detergent-resistant membranes (DRMs) in the plasma membrane. In metabolic-labelling experiments, mature GIPLs and gp63 were transported to DRMs in the plasma membrane with a t(1/2) of 70 and 40 min, respectively. Probably, GIPL transport to the DRMs involves a vesicular mechanism, as transport of both the GIPLs and gp63 was inhibited similarly at 10 degrees C. All GIPL intermediates were quantitatively recovered in Triton X-100 soluble membranes and were largely orientated on the cytoplasmic face of the endoplasmic reticulum, as shown by their sensitivity to exogenous phosphatidylinositol-specific phospho-lipase C. On the contrary, a significant proportion of the mature GIPLs ( approximately 50% of iM4) were accessible to membrane-impermeable probes on the surface of live promastigotes. These results suggest that the GIPLs are flipped across intracellular or plasma membranes during surface transport and that a significant fraction may populate the cytoplasmic leaflet of the plasma membrane. Finally, treatment of L. mexicana promastigotes with myriocin, an inhibitor of sphingolipid biosynthesis, demonstrated that ongoing sphingolipid biosynthesis is not required for the plasma-membrane transport of either gp63 or the GIPLs and that DRMs persist even when cellular levels of the major sphingolipid are depleted by 70%. PMID- 11931668 TI - The Aspergillus niger faeB gene encodes a second feruloyl esterase involved in pectin and xylan degradation and is specifically induced in the presence of aromatic compounds. AB - The faeB gene encoding a second feruloyl esterase from Aspergillus niger has been cloned and characterized. It consists of an open reading frame of 1644 bp containing one intron. The gene encodes a protein of 521 amino acids that has sequence similarity to that of an Aspergillus oryzae tannase. However, the encoded enzyme, feruloyl esterase B (FAEB), does not have tannase activity. Comparison of the physical characteristics and substrate specificity of FAEB with those of a cinnamoyl esterase from A. niger [Kroon, Faulds and Williamson (1996) Biotechnol. Appl. Biochem. 23, 255-262] suggests that they are in fact the same enzyme. The expression of faeB is specifically induced in the presence of certain aromatic compounds, but not in the presence of other constituents present in plant-cell-wall polysaccharides such as arabinoxylan or pectin. The expression profile of faeB in the presence of aromatic compounds was compared with the expression of A. niger faeA, encoding feruloyl esterase A (FAEA), and A. niger bphA, the gene encoding a benzoate-p-hydroxylase. All three genes have different subsets of aromatic compounds that induce their expression, indicating the presence of different transcription activating systems in A. niger that respond to aromatic compounds. Comparison of the activity of FAEA and FAEB on sugar-beet pectin and wheat arabinoxylan demonstrated that they are both involved in the degradation of both polysaccharides, but have opposite preferences for these substrates. FAEA is more active than FAEB towards wheat arabinoxylan, whereas FAEB is more active than FAEA towards sugar-beet pectin. PMID- 11931669 TI - Fusion of two distinct peptide exosite inhibitors of Factor VIIa. AB - Highly potent bifunctional inhibitors of Factor VIIa (FVIIa) were generated by linking two distinct peptides, recently shown to bind to two discrete exosites on the FVIIa protease domain [Dennis, Eigenbrot, Skelton, Ultsch, Santell, Dwyer, O'Connell and Lazarus (2000) Nature (London) 404, 465-470; Dennis, Roberge, Quan and Lazarus (2001) Biochemistry 40, 9513-9521; Roberge, Santell, Dennis, Eigenbrot, Dwyer and Lazarus (2001) Biochemistry 40, 9522-9531]. Fusion peptides consisting of an N-terminal A-series peptide followed by flexible linkers, an E series peptide, and the Z-domain of protein A were expressed in Escherichia coli and purified using IgG-Sepharose affinity chromatography. The fusion peptides were potent anticoagulants and had steep concentration dependence curves in tissue factor-dependent prothrombin time (PT) assays in comparison to the individual peptides or their noncovalent combination. This phenomenon was dependent on the length of the linker joining the A- and E-peptides. The fusion of the peptides increased the extent of inhibition of Factor X (FX) activation to 100% at saturating peptide concentrations, but did not improve the binding affinity for Factor VIIa (FVIIa) at the A- and E- binding sites or the IC(50) for the inhibition of FX activation. Differences between the peptides in the PT fold prolongation in normal and FVII-deficient plasma, in conjunction with the inhibition of (125)I-FVII activation, suggest that the enhanced effects of the fusion peptides involve the inhibition of FVII autoactivation. PMID- 11931670 TI - Phospholipase C-gamma1 is required for cell survival in oxidative stress by protein kinase C. AB - Phospholipase C-gamma1 (PLC-gamma1) activation has been reported to enhance cell survival during the cellular response to oxidative stress. We studied the role of protein kinase C (PKC) pathways in mediating PLC-gamma1 survival signalling in oxidative stress by using mouse embryonic fibroblasts genetically deficient in PLC-gamma1 (Plcg1(-/-)) and its wild type (Plcg1(+/+)). PLC-gamma1 was activated by H(2)O(2) treatment in a dose- and time-dependent manner. Activation of PKC was also markedly increased in both cell lines treated with H(2)O(2) (1-5 mM), but with low doses (50-200 microM), PKC activation was considerably decreased in Plcg1(-/-) cells. After treatment with H(2)O(2), PKC-dependent phosphorylation of Bcl-2 and cell viability of Plcg1(-/-) cells decreased dramatically and caspase-3 like activity increased significantly compared with that of the wild-type cells. Furthermore, pretreatment of Plcg1(+/+) cells with PKC-specific inhibitor decreased levels of PKC-dependent Bcl-2 phosphorylation, enhanced caspase-3 activity and their sensitivity to H(2)O(2). On the contrary, treatment of Plcg1( /-) cells with PKC-specific activator increased the Bcl-2 phosphorylation, decreased caspase-3 activity and improved their survival. These results suggest that PLC-gamma1 mediates survival signalling in oxidative-stress response by PKC dependent phosphorylation of Bcl-2 and inhibition of caspase-3. PMID- 11931671 TI - Probing the serpin structural-transition mechanism in ovalbumin mutant R339T by proteolytic-cleavage kinetics of the reactive-centre loop. AB - A mutant ovalbumin (R339T), but not the wild-type protein, is transformed into the canonical loop-inserted, thermostabilized form after the P1-P1' cleavage [Yamasaki, Arii, Mikami and Hirose (2002) J. Mol. Biol. 315, 113-120]. The loop insertion mechanism in the ovalbumin mutant was investigated by proteolytic cleavage kinetics. The nature of the inserted loop prevented further cleavage of the P1-P1' pre-cleaved R339T mutant by subtilisin, which cleaved the second P8-P7 loop site in the P1-P1' pre-cleaved wild-type protein. After subtilisin proteolysis of the intact R339T, however, two final products that corresponded to the single P1-P1' and double P1-P1'/P8-P7 cleavages were generated with variable ratios depending on the proteolysis conditions. This was accounted for by the occurrence of two mutually competitive reactions: the loop-insertion reaction and the proteolytic cleavage of the second P8-P7 site in the immediate intermediate after the P1-P1' cleavage. The competitive nature of the two reactions enabled us to establish a kinetic method to determine the rate constants of the reactions. The first-order rate constant for the loop insertion was determined to be 4.0 x 10(-3)/s in the R339T mutant. The second-order rate constant for the P8-P7 cleavage in the immediate P1-P1' cleavage product for the R339T mutant was >10 times compared with that for its wild-type counterpart. This highly accessible loop nature may play a crucial role in the loop-insertion mechanism for R339T mutant ovalbumin. PMID- 11931672 TI - A frequent human coagulation Factor VII mutation (A294V, c152) in loop 140s affects the interaction with activators, tissue factor and substrates. AB - Activated Factor VII (FVIIa) is a vitamin-K-dependent serine protease that initiates blood clotting after interacting with its cofactor tissue factor (TF). The complex FVIIa-TF is responsible for the activation of Factor IX (FIX) and Factor X (FX), leading ultimately to the formation of a stable fibrin clot. Activated FX (FXa), a product of FVIIa enzymic activity, is also the most efficient activator of zymogen FVII. Interactions of FVII/FVIIa with its activators, cofactor and substrates have been investigated extensively to define contact regions and residues involved in the formation of the complexes. Site directed mutagenesis and inhibition assays led to the identification of sites removed from the FVIIa active site that influence binding specificity and affinity of the enzyme. In this study we report the characterization of a frequent naturally occurring human FVII mutant, A294V (residue 152 in the chymotrypsin numbering system), located in loop 140s. This region undergoes major rearrangements after FVII activation and is relevant to the development of substrate specificity. FVII A294V shows delayed activation by FXa as well as reduced activity towards peptidyl and macromolecular substrates without impairing the catalytic efficiency of the triad. Also, the interaction of this FVII variant with TF was altered, suggesting that this residue, and more likely loop 140s, plays a pivotal role not only in the recognition of FX by the FVIIa-TF complex, but also in the interaction of FVII with both its activators and cofactor TF. PMID- 11931673 TI - Aminoadipate reductase gene: a new fungal-specific gene for comparative evolutionary analyses. AB - BACKGROUND: In fungi, aminoadipate reductase converts 2-aminoadipate to 2 aminoadipate 6-semialdehyde. However, other organisms have no homologue to the aminoadipate reductase gene and this pathway appears to be restricted to fungi. In this study, we designed degenerate primers for polymerase chain reaction (PCR) amplification of a large fragment of the aminoadipate reductase gene for divergent fungi. RESULTS: Using these primers, we amplified DNA fragments from the archiascomycetous yeast Saitoella complicata and the black-koji mold Aspergillus awamori. Based on an alignment of the deduced amino acid sequences, we constructed phylogenetic trees. These trees are consistent with current ascomycete systematics and demonstrate the potential utility of the aminoadipete reductase gene for phylogenetic analyses of fungi. CONCLUSIONS: We believe that the comparison of aminoadipate reductase among species will be useful for molecular ecological and evolutionary studies of fungi, because this enzyme encoding gene is a fungal-specific gene and generally appears to be single copy. PMID- 11931674 TI - Self-reported and actual physiological responses in social phobia. AB - OBJECTIVES: The aim of the current study was to compare physiological reactions and self-reports of bodily sensations for social phobics, clinically anxious and non-anxious controls across four tasks. DESIGN: A within-group design was used in which each participant took part in four tasks. Two were designed to be demanding, either physically (riding an exercise bicycle) or mentally (mental arithmetic task), while two, a mental imagery task (personally relevant situation) and a social conversation, were designed specifically to be anxiety provoking. METHODS: Of the 54 participants, 18 were generalized social phobics, 18 were clinically anxious but not socially phobic (8 with panic disorder, 6 with generalized anxiety disorder and 4 simple phobics), and 18 were non-anxious. Heart rate, skin conductance, and facial and neck temperatures were recorded continuously during four different tasks and rest periods with corresponding self report ratings of bodily sensations taken to reflect 13 sampling points. RESULTS: There were no group differences on any of the physiological measures during any of the four tasks. However, there were a number of between-group differences with regard to ratings of bodily sensations. Both clinical groups had higher ratings of racing heart than the non-anxious control group during the imagery task. In addition, social phobics had significantly higher ratings of racing heart during the social conversation in relation to both comparison groups. With regard to ratings of body heat, the anxious group had greater ratings than the non-anxious controls during the imagery task. Finally with regard to ratings of sweaty hands, both clinical groups had higher ratings than the non-anxious controls during the social conversation. All three groups were generally inaccurate in their ratings of bodily sensations. PMID- 11931675 TI - Deductive and inductive reasoning in obsessive-compulsive disorder. AB - OBJECTIVES: This study tested the hypothesis that people with obsessive compulsive disorder (OCD) show an inductive reasoning style distinct from people with generalized anxiety disorder (GAD) and from participants in a non-anxious (NA) control group. DESIGN: The experimental procedure consisted of administering a range of six deductive and inductive tasks and a probabilistic task in order to compare reasoning processes between groups. METHODS: Recruitment was in the Montreal area within a French-speaking population. The participants were 12 people with OCD, 12 NA controls and 10 people with GAD. Participants completed a series of written and oral reasoning tasks including the Wason Selection Task, a Bayesian probability task and other inductive tasks, designed by the authors. RESULTS: There were no differences between groups in deductive reasoning. On an inductive "bridging task", the participants with OCD always took longer than the NA control and GAD groups to infer a link between two statements and to elaborate on this possible link. The OCD group alone showed a significant decrease in their degree of conviction about an arbitrary statement after inductively generating reasons to support this statement. Differences in probabilistic reasoning replicated those of previous authors. CONCLUSIONS: The results pinpoint the importance of examining inference processes in people with OCD in order to further refine the clinical applications of behavioural-cognitive therapy for this disorder. PMID- 11931676 TI - Predicting depressive symptoms with a new measure of shame: The Experience of Shame Scale. AB - OBJECTIVE: To investigate whether previous findings from interview studies of a prospective relationship between shame and psychopathology (e.g. Andrews, 1995) could be replicated using questionnaires. DESIGN: A total of 163 university students participated in a longitudinal questionnaire study. METHOD: The Experience of Shame Scale (ESS), a questionnaire based on a previous interview measure, and an established shame scale (TOSCA), were considered in their relation to depressive symptoms assessed at two time points 11 weeks apart. RESULTS: Both scales made significant independent contributions to depressive symptoms at time 1. However, only the ESS predicted additional significant variance in time 2 symptoms when time 1 symptoms were controlled. CONCLUSIONS: It was concluded that the reason for the differential performance of the two scales was that the ESS, like the shame interview, assesses specific areas of shame related to self and performance, whereas the TOSCA assesses general shame and may therefore be more prone to mood-state effects. PMID- 11931677 TI - Death beliefs, superstitious beliefs and health anxiety. AB - OBJECTIVES: The present study explored the association between beliefs about death, superstitious beliefs, and health anxiety. It was hypothesized that negative beliefs about death and superstitious beliefs would be positively correlated with health anxiety. Conversely, positive beliefs about death were hypothesized to be negatively correlated with health anxiety. DESIGN: A cross sectional, correlational and multiple regression design was adopted. METHOD: A sample of 106 Roman Catholics and 197 Atheists completed a questionnaire measuring aspects of health anxiety, spiritual beliefs, and control variables consisting of demographics. RESULTS: Negative beliefs about death and superstitious beliefs were related to health anxiety within both the Roman Catholic and Atheist samples. The expected negative relationship between positive beliefs about death and health anxiety was not supported in either sample. Multiple regression analyses indicated that death beliefs and superstitious beliefs, in combination with background variables, significantly predicted health anxiety in the Roman Catholic sample. For Atheists, although death and superstitious beliefs were identified as significant predictors, when considered with other variables, the additional variance accounted for was not significant. CONCLUSIONS: Negative beliefs about death and superstitious beliefs appear to be positively associated with health anxiety. These types of beliefs may have the potential to offer a useful addition to cognitive-behavioural models of health anxiety. PMID- 11931678 TI - The observed impact of training on competence in clinical supervision. AB - OBJECTIVES: The present study analysed the impact and relative effectiveness of routine training (consultancy) versus routine training plus feedback on clinical supervision. DESIGN: The behaviours of one supervisor and six supervisees were observed longitudinally, and comparisons made between a baseline condition and two subsequent experimental training conditions (with and without feedback) and a maintenance period. METHOD: An observational instrument was used to code N = 1387 interactions between the supervisor and the supervisees. Supervisees' satisfaction with supervision was also recorded longitudinally. RESULTS: The inter-observer reliability was very good initially (K > or = 0.81) and did not "drift". Supervision improved during the experimental phase, but most markedly during the maintenance phase. The results appear to reflect a lag effect for the interventions, which can be most readily explained in terms of a socialization period during which both supervisor and supervisee adapted their styles of interaction. CONCLUSION: Competence in supervision appears to require training. The present methodology affords a promising approach to developing and analysing the effectiveness of supervision. PMID- 11931679 TI - Memory bias against threat in social phobia. AB - OBJECTIVES: The present study examined whether social phobia is an anxiety disorder associated with a memory bias toward threat. DESIGN: Social phobic (N = 16) and non-anxious (N = 17) individuals were compared on their recall of evaluative threat and neutral prose passage content. METHOD: Participants were presented with two evaluative threat and two neutral prose passages and completed an immediate free recall task after each trial. RESULTS: Contrary to expectation, individuals with social phobia recalled a smaller percentage of units from the evaluative threat passages than non-anxious individuals. CONCLUSION: Consistent with the vigilance-avoidance theory, it is suggested that social phobia is an anxiety disorder characterized by the avoidance of elaborate processing of threatening material. PMID- 11931680 TI - A comparison of auditory hallucinations in a psychiatric and non-psychiatric group. AB - OBJECTIVES: To compare auditory hallucinations in a psychiatric and non psychiatric sample. DESIGN: Between-participants group design: a psychiatric group of 14 patients with schizophrenia, and a non-psychiatric group of 16 participants with tinnitus. METHOD: Participants were interviewed using the Mental Health Research Institute Unusual Perceptions Scale (MUPS). RESULTS: The type of hallucinations differed: participants with tinnitus predominantly heard music, whereas the patients with schizophrenia heard voices speaking. The groups also differed in their explanations of the hallucinations. However, there were similarities in contributory factors, emotional responses and coping strategies. CONCLUSIONS: The study highlights the multi-dimensional nature of the auditory hallucinatory experience, and opportunities for intervention. PMID- 11931681 TI - The impact of current media events on hallucinatory content: the experience of the intensive care unit (ICU) patient. AB - OBJECTIVES: To investigate the influence of current media events on hallucinatory content in ICU patients. DESIGN AND METHODS: Patients were interviewed over a one year period, and their descriptions of hallucinatory experiences, together with weekly media stories, were assessed for themes of war. RESULTS: Media coverage for war-related stories rose significantly during the period of war in Kosovo (24/3/99-20/6/99). Patients whose ICU stays coincided with this conflict were more likely to have hallucinatory experiences involving themes of war or the military. Older adults (> 70) were also more susceptible to this effect. CONCLUSIONS: Contextual variables such as current media events can have a significant impact upon the contents of hallucinatory experiences. PMID- 11931682 TI - Brief cognitive screening and self concepts for children with low intellectual functioning. AB - OBJECTIVES: To evaluate the use of brief assessments for children with low intellectual functioning. METHODS: Assessments used the SYSTEMS cognitive screening test and ASK-KIDS self-concept inventory. Participants were children recruited through the hospital and special education classes (N = 39). RESULTS: Results showed that cognitive screening accurately classified children, and their scores correlated closely with full cognitive assessment and achievement. Most children could respond to the self-concept inventory, scales were internally consistent, and profiles were similar to those for children in the normative sample. CONCLUSIONS: Findings support the use of brief assessments of cognitive functioning and self concepts, and suggest further studies with children in clinical settings. PMID- 11931683 TI - Routine cervical cerclage in higher order multiple gestation -- does it prolong the pregnancy? AB - Preterm birth following cervical dilatation is the greatest threat to infants of a multiple pregnancy. Lacking reliable data concerning the effect of prophylactic cerclage, we compared a study group to controls for maternal and perinatal outcome. Sixteen of 94 triplet-, 9 of 18 quadruplet/quintuplet-pregnancies, treated with prophylactic cerclage, were retrospectively compared to those without cervical cerclage respectively. Kruskal-Wallis test and Mann-Whitney-U test were performed as non-parametric one way analysis of variance. For the analysis of frequencies Chi Square test or Fisher's exact test were performed. Odds ratio with 95% confidence interval was used to compare the need for intravenous tocolysis as well as perinatal morbidity and mortality. Gestational age at delivery was not different from the controls in all studied groups. Birth weight revealed a 200 g dominance for the "no cerclage-triplets", while this significant difference was inverted for quadruplets/quintuplets (1245 g vs. 1069 g). With respect to gestational age at birth, need for hospitalisation or medical intervention no benefit was achieved. Moreover, perinatal outcome analysed by arterial pH, APGAR-Score and perinatal mortality was not altered by a prophylactic cerclage. Perinatal morbidity for quadruplets and quintuplets was even higher in cerclage pregnancies. Therefore, these retrospective results disclaim a positive impact of cervical cerclage on pregnancy management or perinatal outcome in multifetal pregnancies. PMID- 11931684 TI - Fundal height as a predictor of early preterm triplet delivery. AB - The purpose of this study was to create graphs of fundal height parameters in triplet pregnancies compared with those in twin pregnancies, and to investigate whether larger fundal heights in triplet pregnancies would predispose them to earlier delivery (before 34 weeks). The subjects were 727 twin pregnant women and 133 triplet pregnant women, who delivered after 1984. Triplet pregnancies showed significantly higher fundal heights compared with twin pregnancies at each gestational age (weeks). In triplet pregnancies, a single fundal height measurement above the 90th percentile before 34 weeks yielded a sensitivity of 31.3% and specificity of 82.4% for delivery before 34 weeks, with a positive predictive value of 50.0% and a negative predictive value of 68.0%. After adjusting for each associated factor using logistic regression, the risk of preterm labour was not significantly associated with a single fundal height measurement above the 90th percentile recorded before 34 weeks. PMID- 11931685 TI - Paternal familial twinning: hypothesis and genetic/medical implications. AB - The phenomenon of paternally dependent familial twinning has been known in human and animal genetics since the 1920s, but still remains without any theoretical explanation and is indeed a neglected field of inquiry. Over the last two decades investigations in reproduction biology have discovered the significant role of multiple paternally dependent errors in fertilization including androgenic triploidy and moles. We suggest the hypothesis that the fathers of twins in the relevant families carry gene variants that increase the probability of dispermy, diplospermy and male pronucleus heterochrony as well as involvement of two male pronuclei in the fertilization of two female meiotic products. Any resulting twins would be an exceptional intermediate between MZ and DZ twins - and might properly be described as "sesquizygotic" (SZ). Paternal familial twinning may also go together with infertility due to triploidy, moles and chimerism. The hypothesis: (i) places the curiosities of paternally derived twinning within the framework of current knowledge of reproductive genetics and verifiable phenomena; (ii) predicts the existence of families in which twinning is associated with reproductive abnormalities; (iii) predicts an occurrence in relevant families of the third and intermediate category of SZ twins. Families with paternal twinning may thus provide the natural selective system for the search of unusual cases of primary chimeras, the frequency of which is still unknown. PMID- 11931686 TI - Heritabilities of apolipoprotein and lipid levels in three countries. AB - This study investigated the influence of genes and environment on the variation of apolipoprotein and lipid levels, which are important intermediate phenotypes in the pathways toward cardiovascular disease. Heritability estimates are presented, including those for apolipoprotein E and AII levels which have rarely been reported before. We studied twin samples from the Netherlands (two cohorts; n = 160 pairs, aged 13-22 and n = 204 pairs, aged 34-62), Australia (n = 1362 pairs, aged 28-92) and Sweden (n = 302 pairs, aged 42-88). The variation of apolipoprotein and lipid levels depended largely on the influences of additive genetic factors in each twin sample. There was no significant evidence for the influence of common environment. No sex differences in heritability estimates for any phenotype in any of the samples were observed. Heritabilities ranged from 0.48-0.87, with most heritabilities exceeding 0.60. The heritability estimates in the Dutch samples were significantly higher than in the Australian sample. The heritabilities for the Swedish were intermediate to the Dutch and the Australian samples and not significantly different from the heritabilities in these other two samples. Although sample specific effects are present, we have shown that genes play a major role in determining the variance of apolipoprotein and lipid levels in four independent twin samples from three different countries. PMID- 11931687 TI - The validity and heritability of self-report osteoarthritis in an Australian older twin sample. AB - In order to investigate the genetic and environmental antecedents of osteoarthritis (OA), self-report measures of joint pain, stiffness and swelling were obtained from a population-based sample of 1242 twin pairs over 50 years of age. In order to provide validation for these self-report measures, a subsample of 118 twin pairs were examined according to the American College of Rheumatology clinical and radiographic criteria for the classification of osteoarthritis. A variety of statistical methods were employed to identify the model derived from self-report variables which would provide optimal prediction of these standardised assessments, and structural equation modelling was used to determine the relative influences of genetic and environmental influences on the development of osteoarthritis. Significant genetic effects were found to contribute to osteoarthritis of the hands, hips and knees in women, with heritability estimates ranging from 30-46% depending on the site. In addition, the additive genetic effects contributing to osteoarthritis in various parts of the body were confirmed to be the same. Statistically significant familial aggregation of osteoarthritis in men was also observed, but it was not possible to determine whether this was due to genetic or shared environmental effects. PMID- 11931688 TI - Ascertainment of a mid-western US female adolescent twin cohort for alcohol studies: assessment of sample representativeness using birth record data. AB - Female twin pairs were identified from birth records, and their families invited to participate in a prospective study of the determinants of alcohol problems in women. We investigated sampling biases arising because of failure to locate families, or non-cooperation of families. Out of 2644 families with a live-born pair (born between July 1975 and December 1986) who survived beyond infancy, contact was established and a brief screening interview completed with 90% (N = 2380). Fewer than 6% of located families declined to participate in the initial screening interview. Predictors of failure to locate a family or to obtain a screening interview were identified from information recorded in birth records, and from neighborhood characteristics identified from 1990 US Census block group data for the family residence when the twins were born. African-American families were under-represented in the final sample, but this effect was barely significant when other variables were controlled for. Under-represented were families where the mother was 19 or younger at the birth of the twins, where the mother herself was born out-of-state, or where information about biological father was not reported in the birth record. Non-participating families on average came from neighborhoods with a higher proportion of residents living in poverty, and with a higher proportion of African-American residents. Sampling biases were however small. The unusual cooperativeness in research of families with twins persists. PMID- 11931689 TI - Estimating two-stage models for genetic influences on alcohol, tobacco or drug use initiation and dependence vulnerability in twin and family data. AB - Genetic research on risk of alcohol, tobacco or drug dependence must make allowance for the partial overlap of risk-factors for initiation of use, and risk factors for dependence or other outcomes in users. Except in the extreme cases where genetic and environmental risk-factors for initiation and dependence overlap completely or are uncorrelated, there is no consensus about how best to estimate the magnitude of genetic or environmental correlations between Initiation and Dependence in twin and family data. We explore by computer simulation the biases to estimates of genetic and environmental parameters caused by model misspecification when Initiation can only be defined as a binary variable. For plausible simulated parameter values, the two-stage genetic models that we consider yield estimates of genetic and environmental variances for Dependence that, although biased, are not very discrepant from the true values. However, estimates of genetic (or environmental) correlations between Initiation and Dependence may be seriously biased, and may differ markedly under different two-stage models. Such estimates may have little credibility unless external data favor selection of one particular model. These problems can be avoided if Initiation can be assessed as a multiple-category variable (e.g. never versus early-onset versus later onset user), with at least two categories measurable in users at risk for dependence. Under these conditions, under certain distributional assumptions, recovery of simulated genetic and environmental correlations becomes possible. Illustrative application of the model to Australian twin data on smoking confirmed substantial heritability of smoking persistence (42%) with minimal overlap with genetic influences on initiation. PMID- 11931690 TI - The personalities of twins: just ordinary folks. AB - Twin studies have demonstrated that personality traits show moderate genetic influence. The conclusions drawn from twin studies rely on the assumptions that twins are representative of the population at large and that monozygotic and dizygotic twins are comparable in every way that might have bearing on the traits being studied. To evaluate these assumptions, we used Multidimensional Personality Questionnaire (MPQ) data from three samples drawn from the Minnesota Twin Registry (totaling 12,971 respondents) to examine the effect sizes associated with mean differences on the 11 MPQ scales and 3 higher-order MPQ factors for singletons versus twins and MZ twins versus DZ twins. The singletons in the samples were family members of the participating twins. We also used ratios of scale variances to examine the significance of variance differences. The only mean or variance difference replicated across all three samples was greater Social Closeness (about.1 standard deviation) for twins than for singletons. This difference was obtained for both males and females. It would appear that, with respect to personality, twins are not systematically different from other people. Our results also highlight the importance of replication in psychological research because each of our large samples showed differences not replicated in other samples. PMID- 11931691 TI - Yoruba customs and beliefs pertaining to twins. AB - The Yoruba are an important ethnic group mainly occupying Southwestern Nigeria. Mainly for genetic reasons, this very large tribe happens to present the highest dizygotic twinning rate in the world (4.4 % of all maternities). The high perinatal mortality rate associated with such pregnancies has contributed to the integration of a special twin belief system within the African traditional religion of this tribe. The latter is based on the concept of a supreme deity called Olodumare or Olorun, assisted by a series of secondary gods (Orisha) while Yoruba religion also involves immortality and reincarnation of the soul based on the animistic cult of ancestors. Twins are therefore given special names and believed to detain special preternatural powers. In keeping with their refined artistic tradition, the Yoruba have produced numerous wooden statuettes called Ibejis that represent the souls of deceased newborn twins and are involved in elaborate rituals. Among Yoruba traditional beliefs and lore some twin-related themes are represented which are also found in other parts of the world. Basic features of the original Yoruba beliefs have found their way into the religious traditions of descendants of African slaves imported in the West Indies and in South America. PMID- 11931692 TI - Twins: not just in science, but in society. PMID- 11931693 TI - Localization of CCTbeta in rat brain and overexpression in insect cells. AB - AIM: To study the localization of CTP: phosphocholine cytidylyltransferase beta isoform (CCTbeta) in rat brain, its expression in insect cells and enzymatic properties. METHODS: Using digoxigenin-labeled CCTbeta probes, in situ hybridization was carried out in rat brain wax sections. CCTbeta was overexpressed in Trichoplusia Ni (Tn) cells using baculovirus expression system. CTP:phosphocholine cytidylyltransferase assay (CT assay) and [3H] metabolic labeling experiment were used to study its activity, properties, and the effect on phosphatidylcholine (PC) synthesis. RESULTS: (1) CCbeta was abundant in CA1, CA2, CA4, and dentate gyrus (DG) region of hippocampus. (2) The content of CCTbeta in transfected Tn cells was over 1 104 times of that in rat brain, and CCTbeta increased the PC synthesis of Tn cells. (3) Hexadecylphosphocholine as well as some ions like Zn2+ and PO3-4 could inhibit the activity of CCTbeta, dCTP was another adaptive substrate of CCTbeta besides CTP. CONCLUSION: CCTbeta showed a similar localization in rat brain with the memory enhancing peptide argipressin (4-8). PMID- 11931694 TI - Reversal of scopolamine-induced spatial memory deficits in rats by TAK-147. AB - AIM: To evaluate effect of TAK-147 on spatial memory deficit induced by scopolamine. METHODS: Morris water maze was used to measure spatial memory in rats and open field test was used to analyse locomotor activity. RESULTS: In the acquisition memory process, scopolamine (0.4 mg/kg, ip) markedly increased the escape latency to the platform. Ip injection of both TAK-147 and donepezil ameliorated scopolamine-induced deficit, dose-related and significant effect was obtained at doses of 0.1-1.0 mg/kg. In the memory retrieval process, increased latency induced by scopolamine (1.5 mg/kg, ip) was also significantly reversed by treatment with TAK-147 (0.1, 0.3, and 1.0 mg/kg), donepezil (0.3 and 1.0 mg/kg), and tacrine (3 and 5 mg/kg), respectively. TAK-147 has a little potent efficacy to donepezil, and was more potent than tacrine. In the locomotor test, both TAK 147 and donepizil created no appreciable change of locomotor activities, compared with scopolamine or saline. CONCLUSION: TAK-147 plays an important role in spatial cognition, and this result provides additional evidence that TAK-147 is an ideal AChE inhibitor and is useful for the treatment of Alzheimer's disease. PMID- 11931695 TI - Effects of histidine, a precursor of histamine, on pentylenetetrazole-induced seizures in rats. AB - AIM: The effect of histidine on pentylenetetrazole-induced seizures was investigated in rats. METHODS: Chemical kindling was elicited by repeated ip injection a subconvulsant dose of pentylenetetrazole (35 mg/kg) once every 48 h until the occurrence of seizure stages 4-5, and seizure activity of kindling was recorded for 30 min. RESULTS: In the kindling development process, ip injection of histidine (200, 500 mg/kg), a precursor of histamine, prolonged latency for the onset of myoclonic jerks and the clonic generalized seizure, and inhibited seizure stage in a dose-dependent manner. In the kindling challenge process, histidine (500, 1000 mg/kg) and H3 antagonist thioperamide (10, 20 microg) al so showed a significant anticonvulsant effect. The inhibitory action of histidine was enhanced significantly by thioperamide (5 microg), however, was antagonized by both alpha fluoromethylhistidine (20 microg), a selective histidine decarboxylase inhibitor and H1 ant agonist pyrilamine (2, 5 mg/kg), dose dependently and significantly. In addition, H2 antagonist zolantidine appeared no appreciable effect, even at a dose of 10 mg/kg. CONCLUSION: These results indicated that brain endogenous histamine may play certain important role in protect against generalized clonic seizures, its action may via presynaptic H3 receptors and postsynaptic H1-receptors. PMID- 11931696 TI - Electrophysiological effects of phytoestrogen genistein on pacemaker cells in sinoatrial nodes of rabbits. AB - AIM: To study the electrophysiological effects of genistein (GST) on pacemaker cells in sinoatrial (SA) nodes of rabbits. METHODS: Parameters of action potential (AP) in SA node were recorded using intracellular microelectrode technique. RESULTS: GST (10 - 150 micromol/L) not only decreased the amplitude of action potential (APA), maximal rate of depolarization (Vmax) [from (6.2 +/- 2.8) to (2.8 +/- 1.4) V/s, P < 0.01], velocity of diastolic (phase 4) depolarization (VDD) [from (55 +/- 14) to (38 +/- 8) mV/s, P < 0.01], and rate of pacemaker firing (RPF) [from (154 +/- 23) to (107 +/- 25) beat/min, P < 0.01], but also prolonged duration of 90 % repolarization of action potential (APD90) in a concentration-dependent manner. Both elevation of calcium concentration (5 mmol/L) in superfusate and application of L-type Ca2+ channel agonist Bay K8644 (0.5 micromol/L) reversed the inhibitory effects of GST on pacemaker cells, while pretreatment with NG-nitro-L-arginine methyl ester (L-NAME, 1 mmol/L), an NO synthase inhibitor, failed to block the electrophysiological effects of GST. CONCLUSION: GST exerted a negative chronotropic action and induced a delayed repolarization of pacemaker cells in SA nodes of rabbits. These effects were likely due to reduction in calcium influx and potassium efflux, but had no association with NO release. PMID- 11931698 TI - Influence of piperacillin on pharmacokinetics of etimicin in healthy volunteers. AB - AIM: To investigate the influence of piperacillin on the pharmacokinetics of etimicin. METHODS: Ten healthy male volunteers subjects were administered randomly with 200 mg of etimicin alone or in combination with 2000 mg of piperacillin. After two weeks washout period, the subjects were crossed over to the second regimen. Blood and urinary samples were collected at specified time intervals. Etimicin concentration was analyzed using micro-bioassay method with Bacillus pumillus as the tested strain. Pharmacokinetic para-meters were determined from serum concentration-time data with the 3p87-software package. RESULTS: Maximum serum concentrations of etimicin alone was (21 +/- 5) mg/L compared with (19 +/- 4) mg/L for the combination. The elimination half-lives were (1.9 +/- 0.4) h and (1.9 +/- 0.2) h for etimicin alone and in combination, respectively. The area under the concentration-time curve of etimicin alone was (38 +/- 7) mg . h . L-1 as opposed to (41 +/- 8) mg . h . L-1 in combination. Twelve hours after administration, urinary recovery rates of etimicin were (56 +/ 8) % and (56 +/- 6) % for etimicin alone and with piperacillin, respectively. CONCLUSION: The results indicated that the pharmacokinetics of etimicin was not affected by concurrent administration of piperacillin in healthy male volunteers. No modification in dosing regimen is necessary when two drugs were co administered. PMID- 11931697 TI - Dauricine-induced changes in monophasic action potentials and effective refractory period of rabbit left ventricle in situ. AB - AIM: To study the effects of dauricine and sotalol on monophasic action potentials and effective refractory period of the rabbit heart in situ. METHODS: Monophasic action potentials recording and programmed electrical stimulation techniques. RESULTS: Iv injection of dauricine 0.5 mg . kg-1 . min-1 for 24 min produced a decrease of the amplitude of monophasic action potentials from (17+/ 6) mV to (7.1 +/- 1.5) mV (P < 0.05) and increased the duration of 50 % and 90 % repolarization of monophasic action potentials and effective refractory period from (130 +/- 26), (167 +/- 25), (128 +/- 12) ms to (198 +/- 33), (235 +/- 34), (185 +/- 25) ms (P < 0.05), respectively. Sotalol had the similar effects on monophasic action potentials and effective refractory period to that of dauricine. Dauricine and sotalol did not change the ratio between effective refractory period and the duration of 90 % repolarization. CONCLUSION: Dauricine and sotalol decreased the amplitude of monophasic action potentials and increased the duration of 50 % and 90 % repolarization and effective refractory period. PMID- 11931699 TI - Pharmacokinetics of multiple intravenous instillation of levofloxacin in Chinese healthy subjects. AB - AIM: To study the pharmacokinetics of multiple doses intravenous infusion of levofloxacin instillation in Chinese healthy volunteers. METHODS: Intravenous infusion of levofloxacin instillation 200 mg within 60 min was given to 10 male healthy volunteers for 7 d, on d 1 and d 7, once-daily, from 2-6 d twice-daily dosing. The concentrations of levofloxacin in serum and urine were assayed by HPLC. RESULTS: The main pharmacokinetic parameters af ter the first dosing were as follows: Cmax was (2.4+/-0.4) mg/L; AUC0- was (16.1 +/- 1.4) mg . h . L-1; T1/2beta was (6.3 +/- 0.3) h. The concentration in serum reached steady state within 72 h. The main parameters after the last dosing were as follows: Cssmax was (2.9 +/- 0.4) mg/L; Cssmin was (0.71 +/- 0.19) mg/L; Cav was (1.40 +/- 0.29) mg/L; AUCss0-12 was (17 +/- 3) mg . h . L-1; T1/2beta was (6.2 +/- 0.8) h. The 24 h cumulative urinary excretion rate was (88 +/- 5) %. From the calculation, the cumulative rate was 1.20; the fluctuation index was 1.30. The difference of T1/2beta and AUC between the first dosing and the last dosing was not significant, and the elimination rate of levofloxacin was not changed after multiple dosing. No clear adverse events were noted during this study. CONCLUSION: There was no accumulation of drug after the repeated intravenous infusion with 200 mg levofloxacin instillation for 7 d. PMID- 11931700 TI - Effect of tachykinins on ascending and descending reflex pathway in rat small intestine. AB - AIM: To examine the effect of tachykinins on the ascending reflex pathway in rat small intestine, we used different selective neurokinin (NK) receptor antagonists (RA): a) NK1-RA: GR-82334 and CP-96.345, b) NK2-RA: MEN-10.376 and L-659.877. The aim was further to investigate the effect of substance P (SP) on the ascending excitatory and descending inhibitory reflex pathway. METHODS: The whole segments of rat ileum (10 cm in length) were studied in an organ bath. Ascending contraction of circle muscle was elicited by anal electrical stimulation (3 Hz, 1 ms, 20 V) and measured as change of intraluminal pressure by a perfused manometric system 2 cm and 4 cm orad of the stimulation site. RESULTS: GR-82334 and CP-96.345 (NK1-RA) caused a significant dose-related inhibition of the oral contraction at a distance of 4 cm: GR-82334 [area: -10 % +/- 8 % (10 nmol/L); -29 % +/ -10 % (1000 nmol/L). P < 0.05, n = 10], CP-96.345 [area: -2 %+/- 6 %(0.1 nmol/L); -14 % +/- 10 % (10 nmol/L). P < 0.01, n = 8], whereas the contractile response at a distance of 2 cm was unaltered (n = 8). In contrast, MEN-10.376 and L-659.877 (NK2-RA) did not alter the amplitude or the area under the curve (n = 10). Neither the NK1- nor the NK2-receptor antagonists had a significant effect on the latency of the reflex response. SP showed a significant increase in the ascending contraction and the descending relaxation (n = 6, P < 0.01). CONCLUSION: These results demonstrate that blockade of NK1-receptors decreases the oral reflex response. Latency of the reflex response remains unchanged, indicating that the effect is not due to an action on interneurons. NK2-receptors do not take part in the ascending reflex in rat small intestine. SP increases the descending relaxant reflex response and ascending contraction. PMID- 11931701 TI - Effect of diet supplementation with l-carnitine on hepatic catabolism of l alanine in rats. AB - AIM: To investigate the effects of chronic supplementation with l-carnitine (LCT) on hepatic catabolism of l-alanine. METHODS: Two groups of male adult Wistar rats were used: 1) supplemented with LCT (1.2 mmol . kg-1 . d-1) dissolved in the drinking water (LCT group) and 2) control group (COG) without LCT supplementation. After one week of LCT supplementation livers from 24 h-fasted rats were perfused in situ and the production of glucose, urea, pyruvate, and l lactate from l-alanine (5 mmol/L) were measured. RESULTS: LCT decreased the production of glucose and urea from l-alanine. In agreement, pyruvate and l lactate production from l-alanine were decreased. However, the supplementation with LCT did not show any significant effect on hepatic glucose production from pyruvate (5 mmol/L) and l-lactate (2 mmol/L). CONCLUSION: LCT supplementation decreased the conversion of l-alanine PMID- 11931702 TI - Effect of phorbol esters on activity of Ha-ras gene promoter in HeLa cells. AB - AIM: To study the role of phorbol esters (PMA) in the activity of promoter of Ha ras in HeLa cells. METHODS: After treatment with PMA, the growth rate of HeLa cells was measured by MTT assay and the expression of Ha-ras gene was detected by reversed transcriptase polymerase chain reaction. The regulation fragment of Ha ras was constructed into the plasmid enhanced yellow fluorescent protein (EYFP) which did not contain promoter element. The recombinant plasmid pRasEYFP was transient transfected to HeLa cells. Using the EYFP gene as reporter, the activity of promoter of Ha-ras after treatment of PMA was assayed. RESULTS: The growth rate of PMA-treated HeLa cells was markedly reduced compared with untreated cells. The expression of Ha-ras gene was obviously decreased and the activity of promoter of Ha-ras was decreased by 34.0 %and 26.7 % respectively in HeLa cells after treated with PMA (100 microg/L) for 48 h and 72 h. In the meanwhile the activity of protein kinase C (PKC) was decreased by 76.3 % and 73.2 % compared with the control cells, respectively. CONCLUSION: PMA play an important role in regulation of the activity of promoter of Ha-ras in HeLa cells. The molecular mechanism may be through the PKC pathway. PMID- 11931703 TI - Myocardial capillary angiogenesis and coronary flow in ischemia tolerance rat by adaptation to intermittent high altitude hypoxia. AB - AIM: To determine the effects of simulated intermittent high altitude hypoxia adaptation (IHA) on coronary capillary and coronary flow (CF) in rat hearts. METHODS: Model of Langendorf-perfused isolated rat hearts were used to measure CF during ischemia-reperfusion, and immunoperoxidase staining assay and computer-aid morphometry analysis were conducted to determine the myocardial capillary densities. Cyclic GMP (cGMP) level in myocardium was measured by radio immunoassay. RESULTS: Pre-ischemia level of CF in IHA rats was higher (IHA28 13.4 mL/min+/-1.5 mL/min, IHA42 15.4 mL/min+/-2.0 mL/min, P < 0.01) than that of normoxic rats (11.0+/-0.8) mL/min, and the recovery of CF after ischemia reperfusion was better in IHA rats. As an adaptive result, the myocardial capillary densities of the left ventricular myocardium in IHA rats were 1.5 times of those in normoxic control rats, but there was no apparent ventricular hypertrophy in IHA rats. Myocardial cGMP content (1.8+/-0.7) nmol/g in IHA rats were increased significantly compared with control rats (1.1+/-0.4) nmol/g, but cGMP level was not altered before and after ischemia-reperfusion in either group. It was also revealed that in isolated rat hearts perfused, myocardial function recovered better in IHA rats than that in normoxic control rats. CONCLUSION: IHA adaptation increased the tolerance of rat hearts against subsequent ischemia reperfusion injury, and increase in coronary circulation and angiogenesis might be the mechanisms of myocardium protected by IHA. PMID- 11931704 TI - Desipramine antagonized corticosterone-induced apoptosis in cultured PC12 cells. AB - AIM: To study possible action mechanism of a tricyclic antidepressant, desipramine (DIM). METHODS: Cultured PC12 cells were exposed to corticosterone in the absence or presence of DIM for 5 d. Agarose gel electrophoresis, flow cytometry, and electron microscopy were used to detect the apoptosis of PC12 cells. RESULTS: Corticosterone 10 micromol/L treatment for 5 d elicited typical apoptotic biochemical and morphological changes including condensed chromatin shaped like crescent moon, nuclear fragmentation, and DNA degradation. The highest percentage of apoptotic cells accumulated to 28 % +/- 9 %. Agarose gel electrophoresis showed typical DNA ladders pattern. While in the presence of DIM 1 or 5 micromol/L, apoptosis percentage was markedly decreased with lightened DNA ladder and ultrastructure of the cells was improved. CONCLUSION: DIM could antagonize the apoptosis in PC12 cells induced by corticosterone, which may be one of the cellular mechanisms of its antidepressant effect. PMID- 11931705 TI - Apoptotic effects of ginsenoside Rh2 on human malignant melanoma A375-S2 cells. AB - AIM: To study the mechanism of ginsenoside-Rh2 (G-Rh2)-induced growth inhibition of A375-S2 cells. METHODS: A375-S2 cell viability and the effect of caspase inhibitors on G-Rh2-induced apoptosis were measured by crystal violet assay. Changes in cellular morphology were observed by phase-contrast microscopy. Apoptosis-specific nucleosomal DNA fragmentation was assayed by agarose gel electrophoresis. Cell cycle distribution was measured by flow cytometry. RESULTS: G-Rh2 inhibited the A375-S2 cell growth in concentration- and time-dependent manners. Caspase family inhibitor, z-Val-Ala-Asp-fluoromethylketone (z-VAD-fmk), caspase-3 inhibitor, z-Asp-Glu-Val-Asp-fluoromethylketone (z-DEVD-fmk), and caspase-8 inhibitor, z-Ile-Glu-Asp-fluoromethylketone (z-IETD-fmk), partially inhibited G-Rh2-induced apoptosis. But caspase-1 inhibitor, Ac-Tyr-Val-Ala-Asp chloromethyl-ketone (Ac-YVAD-cmk), did not antagonize G-Rh2 induced-cell death. CONCLUSION: G-Rh2 suppresses the growth of A375-S2 cells in vitro by inducing apoptosis. G-Rh2-induced apoptosis is partially dependent on caspase-8 and caspase-3 pathway in A375-S2 cells. Other apoptotic pathways might be also related to the induction of apoptosis by G-Rh2. PMID- 11931706 TI - Effect of safrole oxide on vascular endothelial cell growth and apoptosis induced by deprivation of fibroblast growth factor. AB - AIM: To investigate effect of safrole oxide on cell growth and apoptosis induced by deprivation of survival factors (fibroblast growth factors, aFGF and bFGF) in vascular endothelial cells (VEC). METHODS: Morphological changes were observed by light microscopy. Cell growth was determined by MTT (3-[4, 5-dimethylthiazol-2 yl]-2, 5-diphenyltetrazolium) method. DNA fragmentation was analyzed by agarose gel electrophoresis and fluorescence microscopy. Cell cycle distribution was analyzed by flow cytometry (FCM). RESULTS: The cells deprived of FGF were exposed to safrole oxide 5-25 mg/L for 24 h. Cells spreading and growth were promoted (P<0.01), detachment and DNA fragmentation of these cells were suppressed (P<0.01), safrole oxide 10 mg/L had no obvious effect on cell cycle distribution (P>0.05). When the cells were treated with safrole oxide 50-100 mg/L, detachment and DNA fragmentation of VEC were promoted (P<0.01). The cell cycle was blocked at G2-M phase by safrole oxide 100 mg/L. CONCLUSION: Safrole oxide 10 mg/L inhibited, but 100 mg/L promoted apoptosis of VEC. Safrole oxide might be an important compound that affects VEC growth and apoptosis. PMID- 11931707 TI - Expression of intercellular adhesion molecule-1 in U937 foam cells and inhibitory effect of imperatorin. AB - AIM: To investigate the expression level of intercellular adhesion molecule-1 (ICAM-1) in a macrophage-derived foam cell model from human U937 cell line and the inhibitory effect of imperatorin (IMP) on the ICAM-1. METHODS: U937 cells were incubated with oxidized low density lipoprotein (ox-LDL) 80 mg/L for 48 h and a macrophage-derived foam cell model was established. The medium was pretreated with different concentrations of IMP (0, 25, 50, 100 micromol/L). ICAM 1 protein expression in cells was measured with Western blotting; ICAM-1 mRNA level in cells was measured by Northern blotting. RESULTS: After incubated with ox-LDL, ICAM-1 expression level increased greatly. The increase in ICAM-1 protein level and mRNA level was estimated to be about 15-fold and 10-fold. When the cells were pretreated with imperatorin (50, 100 micromol/L), the increase of ICAM 1 in foam cells were remarkably inhibited. Especially when pretreated with IMP 100 micromol/L, the ICAM-1 protein level decreased by 79 % and the mRNA level decreased by 74 % each compared to the level of foam cells. CONCLUSION: After incubated with ox-LDL in vitro, the U937 foam cells showed an enhanced ICAM-1 expression compared with normal U937 cells. IMP could inhibit the expression of ICAM-1 in U937 foam cells. PMID- 11931709 TI - Reduction in monocyte chemoattractant protein-1 mRNA expression in peripheral blood mononuclear cells of diamorphine addicts. AB - AIM: To investigate whether there is a significant difference in the monocyte chemoattractant protein-1 (MCP-1) mRNA expression between diamorphine ( heroin) addicts and normal volunteers. METHODS: Expression of MCP-1 mRNA in the peripheral blood mononuclear cells of diamorphine addicts and normal volunteers was examine d by reverse transcription-polymerase chain reaction (RT-PCR) with beta -actin as a n internal standard. Sequencing of RT-PCR products was performed to confirm the specificity of these products in MCP-1 gene composition. RESULTS: The relative M CP-1 mRNA expression ratios (MCP-1/beta-actin) in the peripheral blood mononuclear cells of normal volunteers control group and diamorphine addicts group were 0.47 +/- 0.12 (n = 15) and 0.21 +/- 0.09 (n =2 1), respectively, and there was a significant difference (P < 0.05). CONCLUSION: The significant reduction of MCP-1 mRNA expression in the peripheral blood mononuclear cells of diamorphine addicts may be one of the mechanisms for the high incidence o f severe infectious diseases, including AIDS, among diamorphine addicts. PMID- 11931708 TI - Inhibitory effects of prostaglandin A1 on apoptosis of rat cardiac microvascular endothelial cells was mediated by NF-kappaB. AB - AIM: To study the effects of prostaglandin A1 (PGA1) on rat cardiac microvascular endothelial cells. METH-ODS: Isolated rat cardiac microvascular endothelial cells were cultured in hypoxia and reoxygen conditions, respectively. Endothelial cell apoptosis was detected by terminal deoxynucleotidyltransferase-mediated dUTP nick end labeling staining. The activity of NF-kappaB was detected by electrophoretic mobility shift assay. Bcl-2 and Bax protein expression were examined by Western blot and bcl-2 mRNA expression was examined by Northern blot. RESULTS: PGA1 reduced endothelial cell apoptosis markedly, inhibited activity of NF-kappaB, and increased expression of Bcl-2 protein and bcl-2 mRNA. However, PGA1 did not alter Bax protein expression resulting in an increase in the ratio of Bcl-2 to Bax. CONCLUSION: PGA1 can inhibit rat cardiac microvascular endothelial cell apoptosis by inhibiting activity of NF-kappaB. PMID- 11931710 TI - Effect of semi-synthesized quercetin water-soluble derivatives on recombinant human phosphatidylinositol 3-kinase p110beta catalytic subunit. AB - AIM: To study the effect of semi-synthesized quercetin water-soluble derivatives sodium quercetin-7-sulfate (SQMS) and disodium quercetin-7,4 -disulfate (SQDS) on recombinant human phosphatidylinositol 3-kinase (PI3-K) p110 beta catalytic subunit. METHODS: Recombinant human PI3-K p110 beta catalytic subunit was expressed by gene engineering. PI3 -K was assayed by incubating recombinant PI3-K p110beta with phosphatidylinositol-4,5-bisphosphate and [gamma-32P]ATP; the [32 P]-radiolabeled lipids were extracted with chloroform and methanol, assessed by thin layer chromatography and visualized by autoradiography. RESULTS: Wortmannin, a specific inhibitor o f PI3-K, showed inhibition on recombinant PI3-K p110beta catalytic subunit in a concentration-dependent manner (2.5 - 20 nmol/L); SQMS and SQD S showed inhibition on recombinant PI3-K p110beta catalytic subunit in a concentration-dependent manner (2.5 - 20 micromol/L). CONCLUSION: Semi synthesized quercetin water-soluble derivatives were a type of inhibitors of PI3 K. The recombinant PI3-K p110beta catalytic subunit might be used as a molecular target for simpler filtrating and development of more effective inhibitors of PI3 K. PMID- 11931711 TI - Immunomodulatory activity of orphanin FQ/nociceptin on traumatic rats. AB - AIM: To explore the neuro-immune modulatory effect of orphanin FQ/nociceptin (OFQ) and opioid receptor like 1 (ORL1) receptor on the traumatic rats. METHODS: The quantitative method of immuno-cytochemistry and i n situ hybridization combined with cytokine bioassay were used to detect the expression of endogenous OFQ and ORL1 and the production of interleukin-1 (IL-1) and tumor necrosis factor alpha (TNF-alpha) from peritoneal macrophage. RESULTS: Strong signals for both OFQ immuno-reactive cells and ORL1 mRNA were detected in cerebral cortex, hippocampus, and hypothalamus in normal condition, whereas they were significantly reduced after trauma (P<0.05). However, the production of IL-1 and TNF-alpha from peritoneal macrophage was increased, when expressed as percentage of enhancement, the increment attained to 233 % and 521 % (sample dilution 1:4), 195 % and 566 % (1:8), 233 % and 757 % ( 1:16), 214 % and 622 % (1:32), respectively, after trauma. After icv injection of OFQ at doses of 0.055 nmol, 0.55 nmol, and 2.75 nmol, the units of IL-1 and TNF-alpha were reversed (P<0.05); however, the action of OFQ (0.55 nmol) was blocked by ORL1 selective antagonist [phe1psi(CH2-NH)Gly2]nociceptin-(1-13)-NH2. CONCLUSION: OFQ and ORL1, the new opioid peptide system, are involved in the immune response elicited by traumatic stress. PMID- 11931712 TI - Role of statin drugs in acute coronary syndromes. PMID- 11931713 TI - Searching for the mountains of the moon: genome scans for atherosclerosis. AB - Several research groups have begun mounting large, ambitious family studies to map genes for atherosclerosis, heart disease, and their major risk factors using whole genome linkage and/or disequilibrium scans. Some of the problems, pitfalls, and challenges of this exciting effort are examined and illustrated with lessons from an earlier mapping problem. PMID- 11931714 TI - Genetics of eating and its relation to obesity. AB - Humans show remarkable variability in their eating styles. Reasons for this variability are poorly understood, but carry important implications for the onset of obesity and its comorbidities. Although it is well established that genes influence obesity, genetic influences on human eating patterns have received less attention. Dramatic increases in the prevalence of obesity justify additional research into the genetics of eating style. This review highlights what is known about the genetics of food preferences and selection in humans. Data from family and twin studies suggest a moderate genetic contribution to eating style, with clear environmental effects emerging as well. Select studies show support for gene-environment interactions and common genetic bases for certain dietary patterns, obesity, and/or metabolic complications. Genetically informative designs that infuse innovative behavioral measures may help explain individual differences in eating styles and hopefully promote more effective obesity treatments. PMID- 11931716 TI - Genetic determinants of diabetes and atherosclerosis. AB - Common risk factors for type 2 diabetes and cardiovascular disease are suspected because of the higher than expected prevalence of cardiovascular disease in individuals with newly diagnosed type 2 diabetes and in nondiabetic individuals with impaired glucose tolerance. Some of these risk factors may be genetic in origin. The risk of diabetes and cardiovascular disease may be linked through common pathways, including insulin resistance and/or inflammation. In this review, we describe the rationale for proposing these as common mechanisms affecting both diabetes and cardiovascular risk, and we then summarize the available evidence suggesting that common genes within these pathways have pleiotropic effects influencing susceptibility to both diseases. PMID- 11931715 TI - Pharmacogenetics of Lipid-lowering Therapies. AB - Cardiovascular disease is associated with nonmodifiable risk factors such as age, gender, and genetic background, and with modifiable risk factors such as lipid concentrations. Lowering serum lipid levels has been demonstrated to slow the progression of, or even induce regression in, atherosclerosis. However, like any other drug treatment, the magnitude of plasma lipid responses to drug therapies varies considerably among individuals. Pharmacogenetics provides the experimental basis to understand the variability in response to drugs as a function of the individual genetic makeup. Information from small clinical trials reveals that several candidate genes may hold some promise in our quest to predict individual success to hypolipemic drug treatment. However, the current clinical relevance of this knowledge is quite limited due to the small effects observed for each of the genetic markers examined. Future progress in this area will be driven by studying gene-gene and gene-treatment interactions in much larger patient populations. PMID- 11931718 TI - Extending knowledge of the benefits of statins: The Heart Protection Study. PMID- 11931717 TI - Recent advances in liver-directed gene therapy for dyslipidemia. AB - As currently available preventive and therapeutic interventions for hypercholesterolemia are ineffective in a substantial proportion of patients, severe dyslipidemias associated with atherosclerotic vascular disease remain an important target for the development of novel gene therapies. The development of a safe and efficient gene transfer vector has been a major challenge in liver directed gene therapy, but recently significant progress has been made in this area. Proof-of-principle experiments indicate that the transfer of lipid modifying genes to the liver is an effective method to restore normal plasma lipids and protect against atherosclerosis. This article summarizes recent developments in liver-directed gene delivery and reviews data on the treatment of dyslipidemias and prevention of atherosclerosis in animals. The evidence presented suggests that some of the approaches taken in animals may be ready for clinical trials in the near future. PMID- 11931719 TI - Orphan nuclear receptors find a home in the arterial wall. AB - Orphan nuclear receptors of the peroxisome proliferator activated receptor (PPAR) and liver X receptor (LXR) subfamilies have been shown to play critical roles in both local and systemic lipid metabolism. The PPARs control fatty acid metabolism in various cell types, including adipocytes, liver, and macrophages. The LXRs have been implicated in the regulation of cholesterol metabolism in the liver, intestines, and macrophages. The importance of these receptors in physiologic lipid metabolism suggests that they may influence the development of metabolic disorders such as obesity, diabetes, and atherosclerosis. Furthermore, the ability of these receptors to be modulated pharmacologically makes them attractive therapeutic targets. This review focuses on the role of PPAR and LXR signaling pathways in macrophage lipid metabolism and the potential of these pathways to modulate the development of atherosclerosis. PMID- 11931720 TI - Mechanisms of abdominal aortic aneurysm formation. AB - Abdominal aortic aneurysms (AAA) are permanent dilations in the infra-renal area in which the arterial tissue is characterized by inflammation and medial degeneration. AAAs are a common vascular disorder and cause significant mortality in the aged population. Despite the high prevalence of this disease, there is limited knowledge on the mechanisms responsible for the vascular pathology. Therefore, current therapeutic options are restricted to surgical intervention and are predicated on the assumed propensity for rupture as the vessel enlarges. Current research focuses on inflammatory processes and their role in proteolytic degradation of the elastin and collagen fibers of the vessel wall. Definition of specific mechanisms would identify target sites for potential pharmacologic intervention and markedly improve the medical treatment and prognosis of AAA. PMID- 11931722 TI - Animal models of spontaneous plaque rupture: the holy grail of experimental atherosclerosis research. AB - Throughout the history of atherosclerosis research we have sought animal models of the disease process that exhibit high frequencies of the features that make human plaque a clinical risk: plaque rupture, mural thrombosis, and intra-plaque hemorrhage. This type of model is needed to determine the mechanisms by which plaques rupture and to design and test therapeutic interventions for stabilizing plaques. Studies of domestic and exotic animals have shown that most species will spontaneously develop fatty streaks and in some cases atheromatous lesions with sufficient time, but that rupture and thrombosis is exceedingly rare. Even with addition of fat and cholesterol to the diet, lesion development is accelerated but does not increase the frequency with which plaques rupture in most animal models. However, recently we have observed high frequencies of intra-plaque hemorrhage in the innominate/brachiocephalic arteries of older, chow-fed, hyperlipidemic, apolipoprotein E-deficient mice, and high frequencies of plaque rupture with mural thrombus in younger apolipoprotein E-deficient mice fed a high fat diet. This suggests that plaque rupture and secondary thrombosis are frequent and reproducible occurrences at specific sites in apolipoprotein E-deficient mice, and that the timing and pathobiology of the ruptures are influenced by lipid status in this murine model. PMID- 11931721 TI - Receptor for advanced glycation endproducts (RAGE) and vascular inflammation: insights into the pathogenesis of macrovascular complications in diabetes. AB - The incidence and severity of atherosclerosis is increased in patients with diabetes. Indeed, accelerated macrovascular disease in diabetic patients has emerged as a leading cause of morbidity and mortality in the United States and worldwide. Multiple investigations have suggested that there are numerous potential contributory factors that underlie these observations. Our laboratory has focused on the contribution of receptor for advanced glycation endproducts (RAGE) and its proinflammatory ligands, advanced glycation endproducts (AGEs) and S100/calgranulins in vascular perturbation, manifested as enhanced atherogenesis or accelerated restenosis after angioplasty. In rodent models of diabetic complications, blockade of RAGE suppressed vascular hyperpermeability, accelerated atherosclerotic lesion area and complexity in diabetic apolipoprotein E-deficient mice, and prevented exaggerated neointimal formation in hyperglycemic fatty Zucker rats subjected to injury of the carotid artery. In this review, we summarize these findings and provide an overview of distinct mechanisms that contribute to the development of accelerated diabetic macrovascular disease. Insights into therapeutic strategies to prevent or interrupt these processes are presented. PMID- 11931724 TI - Painful Peripheral Neuropathy. AB - Treatment of neuropathic pain is the primary focus of management for many patients with painful peripheral neuropathies. Neuropathic pain is a common feature of many peripheral neuropathies including those associated with diabetes, uremia, HIV infection, and alcohol abuse. Pain is also present in the majority of patients with idiopathic sensory and sensorimotor polyneuropathies. A growing number of pharmacologic agents are available for the treatment of neuropathic pain. The medications that have undergone the most rigorous study are the tricyclic antidepressants and anticonvulsants. These two families of medications are widely used and represent first-line agents in the management of neuropathic pain. Pain management should begin with a concerted effort to identify the etiology of the neuropathy, as directed therapy may help alleviate the symptoms. When initiating pharmacotherapy for neuropathic pain, one must individualize treatment and choose an agent that is likely to be tolerated, as adverse events are not uncommon for some of the medications. Treatment of neuropathic pain remains challenging, with considerable variability in an individual's response to the various agents and even to different drugs in the same class. However, monotherapy with a well-chosen agent or rational polypharmacy that combines medications with different mechanisms of action will benefit a majority of patients with neuropathic pain. PMID- 11931723 TI - ATP-binding cassette (ABC) transporters in atherosclerosis. AB - Macrophages play a central role in the initiation and progression of atherosclerotic lesions. In the nascent lesion, macrophages transform into foam cells through the excessive accumulation of cholesteryl esters. Dysfunctional lipid homeostasis in macrophages and foam cells ultimately results in the breakdown of membrane integrity and cell death. Studies within the past 2 years have implicated a defined subset of multispan transmembrane proteins, the ATP binding cassette (ABC) transporters, in macrophage lipid homeostasis. The recent finding that ABCA1, beyond its function as a major regulator of plasma high density lipoprotein metabolism, exerts significant antiatherosclerotic activities has provided the first direct evidence for the role of an ABC transporter in the pathogenesis of atherosclerosis. PMID- 11931725 TI - Peripheral Neuropathy Due to Leprosy. AB - Leprosy, also known as Hansen's disease, remains a significant cause of disability worldwide. After the introduction of treatment regimens using a combination of dapsone, rifampin, and clofazimine, the prevalence of the disease declined from 5.4 million registered cases in 1985 to less than a million in 1999. However, the incidence of new cases has remained stable due at least in part to a population of asymptomatic carriers. Immune-mediated nerve damage can occur during treatment or after treatment is completed and mandates continued careful follow-up of patients. Patient education and rehabilitation are crucial aspects of disease management and prevention of disability. In the US, patient care and medications are available through regional clinics sponsored by the Department of Health and Human Services. Patients should contact the National Hansen's Disease Program at 1770 Physician's Park Drive, Baton Rouge, Louisiana 70816; 1-800-642-2477. PMID- 11931726 TI - Peripheral Neuropathy Due to Cobalamin Deficiency. AB - Vitamin B12, or cobalamin (Cbl), deficiency can produce a number of neurologic complications, including myelopathy, peripheral neuropathy, optic neuropathy, and dementia. The myelopathy, combined systems disease, is probably the most well known manifestation, and is usually readily recognized. The frequency with which peripheral neuropathy is the sole presenting feature of Cbl deficiency is a point of controversy. The prevalence and the clinical and electrophysiologic features of Cbl deficiency peripheral neuropathy have not been well characterized. In addition, there is evidence that the commonly used assays of serum Cbl are not adequately sensitive. Testing the serum metabolites methylmalonic acid and homocysteine can increase the identification of Cbl deficient patients. Treatment with parenteral Cbl injections may not produce improvement of neurologic deficits, but might prevent worsening. In some patients with Cbl deficiency, oral Cbl may be an effective therapy. PMID- 11931728 TI - Human Immunodeficiency Virus (HIV) in Children. AB - Recent advances in antiretroviral treatment have changed the landscape of pediatric HIV--not only has perinatal treatment markedly diminished HIV transmission rates, but highly active antiretroviral treatments (HAART) have, with few exceptions, transformed HIV into a chronic nonfatal disorder. Antiretroviral therapies are complex and fraught with potential for side effects. The neurologist involved with HIV-infected children should have a firm grasp on the different therapeutic agents used among infected children, especially as regards drug metabolism and indications for treatment. PMID- 11931727 TI - Thymectomy for Myasthenia Gravis. AB - No Class I studies of the effectiveness of thymectomy for myasthenia gravis (MG) have been performed. Most Class II studies comparing outcomes in MG patients with and without thymectomy demonstrated higher MG remission and improvement rates in patients undergoing thymectomy. However, these Class II studies were also consistently confounded by important differences between MG patients in surgical and nonsurgical groups. Myasthenia gravis patients undergoing thymectomy were younger, more often women and were more likely to have severe myasthenia. The authors of this paper cannot determine from these Class II studies whether the observed association between thymectomy and improved MG outcomes was a result of a thymectomy benefit or was merely a result of the multiple differences in baseline characteristics between the surgical and nonsurgical groups. The authors concluded that the benefit of thymectomy in non-thymomatous autoimmune MG has not been conclusively established. Thus, for patients with non-thymomatous autoimmune myasthenia gravis, thymectomy should only be considered an option to increase the probability of remission or improvement. The quality standards subcommittee of the American Academy of Neurology recently adopted this position. PMID- 11931729 TI - Congenital Cytomegalovirus Infection. AB - Intrauterine infection with cytomegalovirus (CMV), a betaherpesvirus, remains the most frequent congenital virus infection in many regions of the world. Although most CMV-infected newborns lack signs of CMV infection, approximately 10% have signs that can consist of low birth weight, jaundice, hepatosplenomegaly, skin rash, microcephaly, and chorioretinitis. Neonates with signs of CMV infection at birth have high rates of audiologic and neurodevelopmental sequelae. Although postnatal therapy with ganciclovir transiently reduces virus shedding and may lessen the audiologic consequences of CMV in some infected infants, additional strategies are needed to prevent congenital CMV disease and to improve the neurodevelopmental prognosis of infants infected with CMV in utero. Some cases of intrauterine infections can be prevented in susceptible women by avoiding contact with the urine or saliva of young children who may be shedding CMV. Vaccines against CMV remain in the experimental stages of development. Termination of pregnancy can be offered to women whose infants have evidence of intrauterine CMV infection and sonographic signs of central nervous system damage. Infants who survive symptomatic intrauterine infections have high rates of neurodevelopmental sequelae and require comprehensive evaluation and therapy through center and home based early intervention programs. PMID- 11931730 TI - Herpes Simplex Virus in Children. AB - Herpes simplex virus (HSV) infections are ubiquitous. Children are infected with HSV resulting in totally asymptomatic acquisition to life-threatening disease. Therapy of HSV diseases of children can be considered according to severity and time of acquisition. Neonatal herpes simplex virus infections take one of three forms--disease localized to skin, eye, or mouth (SEM), encephalitis, or multiorgan disseminated disease. Treatment consists of intravenous (IV) administration of acyclovir. Supportive care for patients with life-threatening disease is an integral component of patient management. Mucocutaneous HSV infections in the immunocompromised host can be treated with either intravenous acyclovir or one of the orally bioavailable antiviral therapies. For hospitalized patients, therapy consists of IV acyclovir at 5 mg/kg every 8 hours for 7 to 14 days. For ambulatory patients, therapy is tailored according to age. For children less than 12 years of age, oral acyclovir is administered at a dosage of 20 mg/kg every eight hours. Although no controlled studies have been performed with valaciclovir or famciclovir, the pharmacokinetics of these medications would suggest superiority over acyclovir. Dosage recommendations have not been established for young children. For postpubertal children, dosage should mirror that of adults. Valaciclovir is administered at 500 mg twice daily. Famciclovir is administered at 125 mg three times daily. Herpes simplex keratoconjunctivitis is treated with topical triflurothymidine. Two drops are applied to the infected eye five times daily until resolved. Recurrences are managed in a similar manner. Some physicians administer oral acyclovir at the doses noted above in order to prevent frequent recurrences. Genital HSV infections can be treated with acyclovir, valaciclovir, or famciclovir. Episodic treatment of recurrent episodes is usually not necessary in childhood. Importantly, all data on the use of these compounds for these conditions have been generated in adults. Physician judgment is required for the management of recurrent herpes labialis, erythema multiforme, and herpes gladitorum. No controlled studies have been performed in children, although experience with acyclovir, valaciclovir, and famciclovir have resulted in their use. PMID- 11931731 TI - Meningitis in the Neonate. AB - Group B beta-hemolytic streptococci and Escherichia coli strains account for approximately two thirds of all cases of neonatal meningitis, while bacteria that typically account for meningitis in older age groups (Haemophilus influenzae type B, Neisseria meningitidis, and Streptococcus pneumoniae) are infrequent causes of meningitis in the neonatal population. As with other medical problems in neonates, signs and symptoms of bacterial infection of the central nervous system are generally few in number and nonspecific in nature. Manifestations that can suggest meningitis, as well as other serious illnesses, include temperature instability, lethargy, respiratory distress, poor feeding, vomiting, and diarrhea. Signs suggestive of meningeal irritation, including stiff neck, bulging fontanelle, convulsions, and opisthotonus, occur only in a minority of neonates with bacterial meningitis and cannot be relied on solely to identify such patients. Ampicillin and either gentamicin or cefotaxime are recommended for initial empiric therapy of neonatal meningitis. When the results of the cerebrospinal fluid (CSF) culture and susceptibilities are known, therapy can be narrowed to cover the specific pathogen identified. In general, penicillin G or ampicillin is preferred for group B streptococcal meningitis, ampicillin for Listeria monocytogenes meningitis, and ampicillin plus either an aminoglycoside or cefotaxime for gram-negative meningitis. For the very low birth weight neonate who has been in the nursery for a prolonged period of time, organisms such as enterococci and gentamicin-resistant gram-negative enteric bacilli must also be considered. In patients with long-term vascular catheters, Staphylococcus aureus or coagulase-negative staphylococci must also be considered. Empiric combinations of antibiotics for such patients would include ampicillin or vancomycin, plus amikacin or cefotaxime. All neonates should undergo repeat CSF examination and culture at 48 to 72 hours after initiation of therapy. If organisms are observed on gram stain, modification of the therapeutic regimen should be considered, and neuroimaging should be performed. In general, therapy should be continued for 14 to 21 days for neonatal meningitis caused by group B streptococci or L. monocytogenes, and for at least 21 days for disease caused by gram-negative enteric bacilli. All patients with neonatal meningitis should have hearing and development monitored serially. The first audiologic evaluation should occur 4 to 6 weeks after resolution of the meningitis. PMID- 11931732 TI - Tuberculous Meningitis in Children. AB - Initial empiric treatment for central nervous system (CNS) tuberculosis should include four antituberculous drugs until results of cultures and sensitivities are available. Treatment should include isoniazid, rifampin, pyrazinamide, and either ethambutol or streptomycin. Total treatment should extend for 12 months. Daily therapy should be used for the first 2 months, followed by either twice a week treatment or continued with daily therapy for the duration with directly observed therapy (DOT). Pyrazinamide should be included in all treatment regimes for the first 2 months of therapy. Corticosteroids should be used in the management of children with tuberculous meningitis. Corticosteroids have been shown to decrease mortality, long-term neurologic complications, and permanent sequelae. Prednisone is often used at a dosage of 1 to 2 mg/kg per day. Steroids should be used for 4 to 6 weeks, and then tapered over the next 2 to 3 weeks. Cerebrospinal fluid (CSF) cultures and other infected sites must be aggressively pursued in order to obtain an organism for identification and sensitivities testing. Cranial CT scans with contrast should be included in the early diagnostic work-up of a child with suspected CNS tuberculosis infection. Hydrocephalus is often an early finding and may be helpful in establishing the diagnosis of CNS tuberculosis. Treatment of CNS tuberculosis should be for 12 months. All children with CNS tuberculosis should be promptly reported to the local public health department. Public health will facilitate the case-contact study and assist with follow-up and DOT after discharge. Directly observed therapy should be given for the entire treatment course. This is best accomplished with the collaboration of local public health services. Children with tuberculous meningitis should be evaluated in follow-up monthly. Monitoring should include determining adherence to drug treatment, an interval history for signs and symptoms of disease progression, careful physical examinations and evaluation for adverse effects of drugs. Liver function tests should be obtained at baseline, 2-, 4-, 6-, and 8 weeks, and then monthly for the first several months of treatment. Children with tuberculous meningitis should be tested for HIV infection, including pre- and post-test counseling. PMID- 11931733 TI - Getting a GRIP on liprins. AB - Two papers in this issue of Neuron add a new dimension to our understanding of liprin and LAR RPTP function during synapse formation. present evidence that Dliprin-alpha interacts with Dlar to regulate presynaptic morphogenesis of the Drosophila neuromuscular junction. demonstrate that mammalian liprin-alpha1 forms a complex with the PDZ protein GRIP and LAR in both pre- and postsynaptic compartments of hippocampal neurons and is required for the clustering of the GluR2 glutamate receptor in dendritic spines. PMID- 11931734 TI - Adaptation through translocation. AB - Papers by and, in this issue of Neuron, describe how massive and rapid translocation of specific elements of the phototransduction cascades in different phyla, namely, the G protein (transducin) in vertebrate rods and light-sensitive TRPL channels in the microvillar rhabdomeres of Drosophila, contribute to photoreceptor adaptation. PMID- 11931735 TI - Action potentials that go the distance. AB - Dendrodendritic inhibition between mitral and granule cells in the olfactory bulb is thought to play an important role in olfactory discrimination. In this issue of Neuron, explore the propagation of action potentials along the secondary dendrites of mitral cells and their modulation by dendrodendritic inhibition. PMID- 11931736 TI - Should bad workmen always blame their tools? AB - In this issue of Neuron, differentiated brain regions in humans that are selectively responsive to viewing motion: (1) of humans, (2) of tools/utensils, and (3) in general. Active regions were the superior temporal sulcus, middle temporal gyrus, and MT/V5, respectively. This study provides impetus for future work in motion perception and its relationship to apraxia. PMID- 11931737 TI - Prime movers of synaptic vesicle exocytosis. AB - Chemical synapses contain specialized pre- and postsynaptic structures that underlie rapid synaptic transmission and its modulation. Studies of postsynaptic organization have revealed a network of interacting proteins that enable rapid synaptic responses and their modulation. Recent genetic and electrophysiological studies on two active zone proteins-RIM and Munc13-reveal important roles in priming vesicles for Ca(2+)-triggered fusion and in mediating the regulation of this process. This work sheds new light on how presynaptic structure provides speed and plasticity to synaptic transmission. PMID- 11931738 TI - Neurobiology of depression. AB - Current treatments for depression are inadequate for many individuals, and progress in understanding the neurobiology of depression is slow. Several promising hypotheses of depression and antidepressant action have been formulated recently. These hypotheses are based largely on dysregulation of the hypothalamic pituitary-adrenal axis and hippocampus and implicate corticotropin-releasing factor, glucocorticoids, brain-derived neurotrophic factor, and CREB. Recent work has looked beyond hippocampus to other brain areas that are also likely involved. For example, nucleus accumbens, amygdala, and certain hypothalamic nuclei are critical in regulating motivation, eating, sleeping, energy level, circadian rhythm, and responses to rewarding and aversive stimuli, which are all abnormal in depressed patients. A neurobiologic understanding of depression also requires identification of the genes that make individuals vulnerable or resistant to the syndrome. These advances will fundamentally improve the treatment and prevention of depression. PMID- 11931739 TI - Drosophila liprin-alpha and the receptor phosphatase Dlar control synapse morphogenesis. AB - Here, we examine the synaptic function of the receptor protein tyrosine phosphatase (RPTP), Dlar, and an associated intracellular protein, Dliprin-alpha, at the Drosophila larval neuromuscular junction. We show that Dliprin-alpha and Dlar are required for normal synaptic morphology. We also find that synapse complexity is proportional to the amount of Dlar gene product, suggesting that Dlar activity determines synapse size. Ultrastructural analysis reveals that Dliprin-alpha and Dlar are required to define the size and shape of the presynaptic active zone. Accordingly, there is a concomitant decrease in synaptic transmission in both mutants. Finally, epistasis analysis indicates that Dliprin alpha is required for Dlar's action at the synapse. These data suggest a model where Dliprin-alpha and Dlar cooperate to regulate the formation and/or maintenance of a network of presynaptic proteins. PMID- 11931740 TI - Interaction between GRIP and liprin-alpha/SYD2 is required for AMPA receptor targeting. AB - Interaction with the multi-PDZ protein GRIP is required for the synaptic targeting of AMPA receptors, but the underlying mechanism is unknown. We show that GRIP binds to the liprin-alpha/SYD2 family of proteins that interact with LAR receptor protein tyrosine phosphatases (LAR-RPTPs) and that are implicated in presynaptic development. In neurons, liprin-alpha and LAR-RPTP are enriched at synapses and coimmunoprecipitate with GRIP and AMPA receptors. Dominant-negative constructs that interfere with the GRIP-liprin interaction disrupt the surface expression and dendritic clustering of AMPA receptors in cultured neurons. Thus, by mediating the targeting of liprin/GRIP-associated proteins, liprin-alpha is important for postsynaptic as well as presynaptic maturation. PMID- 11931741 TI - NSF ATPase and alpha-/beta-SNAPs disassemble the AMPA receptor-PICK1 complex. AB - AMPA receptor (AMPAR) trafficking is crucial for synaptic plasticity that may be important for learning and memory. NSF and PICK1 bind the AMPAR GluR2 subunit and are involved in trafficking of AMPARs. Here, we show that GluR2, PICK1, NSF, and alpha-/beta-SNAPs form a complex in the presence of ATPgammaS. Similar to SNARE complex disassembly, NSF ATPase activity disrupts PICK1-GluR2 interactions in this complex. Alpha- and beta-SNAP have differential effects on this reaction. SNAP overexpression in hippocampal neurons leads to corresponding changes in AMPAR trafficking by acting on GluR2-PICK1 complexes. This demonstrates that the previously reported synaptic stabilization of AMPARs by NSF involves disruption of GluR2-PICK1 interactions. Furthermore, we are reporting a non-SNARE substrate for NSF disassembly activity. PMID- 11931742 TI - Drosophila CLOCK protein is under posttranscriptional control and influences light-induced activity. AB - In the Drosophila circadian clock, daily cycles in the RNA levels of dclock (dClk) are antiphase to those of period (per). We altered the timing/levels of dClk expression by generating transgenic flies whereby per circadian regulatory sequences were used to drive rhythmic transcription of dClk. The results indicate that posttranscriptional mechanisms make substantial contributions to the temporal changes in the abundance of the dCLK protein. Circadian regulation is largely unaffected in the transgenic per-dClk flies despite higher mean levels of dCLK. However, in per-dClk flies the duration of morning activity is lengthened in light-dark cycles and light pulses evoke longer lasting bouts of activity. Our findings suggest that, in addition to a role in generating circadian rhythms, dCLK modulates the direct effects of light on locomotion. PMID- 11931743 TI - Light-regulated subcellular translocation of Drosophila TRPL channels induces long-term adaptation and modifies the light-induced current. AB - Drosophila phototransduction results in the opening of two classes of cation channels, composed of the channel subunits transient receptor potential (TRP), TRP-like (TRPL), and TRPgamma. Here, we report that one of these subunits, TRPL, is translocated back and forth between the signaling membrane and an intracellular compartment by a light-regulated mechanism. A high level of rhabdomeral TRPL, characteristic of dark-raised flies, is functionally manifested in the properties of the light-induced current. These flies are more sensitive than flies with no or reduced TRPL level to dim background lights, and they respond to a wider range of light intensities, which fit them to function better in darkness or dim background illumination. Thus, TRPL translocation represents a novel mechanism to fine tune visual responses. PMID- 11931744 TI - Massive light-driven translocation of transducin between the two major compartments of rod cells: a novel mechanism of light adaptation. AB - We report a new cellular mechanism of rod photoreceptor adaptation in vivo, which is triggered by daylight levels of illumination. The mechanism involves a massive light-dependent translocation of the photoreceptor-specific G protein, transducin, between the functional compartments of rods. To characterize the mechanism, we developed a novel technique that combines serial tangential cryodissection of the rat retina with Western blot analysis of protein distribution in the sections. Up to 90% of transducin translocates from rod outer segments to other cellular compartments on the time scale of tens of minutes. The reduction in the transducin content of the rod outer segments is accompanied by a corresponding reduction in the amplification of the rod photoresponse, allowing rods to operate in illumination up to 10-fold higher than would otherwise be possible. PMID- 11931745 TI - Metabotropic-mediated kainate receptor regulation of IsAHP and excitability in pyramidal cells. AB - Kainate receptors (KARs) on CA1 pyramidal cells make no detectable contribution to EPSCs. We report that these receptors have a metabotropic function, as shown previously for CA1 interneurons. Brief kainate exposure caused long-lasting inhibition of a postspike potassium current (I(sAHP)) in CA1 pyramidal cells. The pharmacological profile was independent of AMPA receptors or the GluR5 subunit, indicating a possible role for the GluR6 subunit. KAR inhibition of I(sAHP) did not require ionotropic action or network activity, but was blocked by the inhibitor of pertussis toxin-sensitive G proteins, N-ethylmaleimide (NEM), or the PKC inhibitor calphostin C. These data suggest how KARs, putatively containing GluR6, directly increase excitability of CA1 pyramidal cells and help explain the propensity for seizure activity following KAR activation. PMID- 11931746 TI - Dynamic gating of spike propagation in the mitral cell lateral dendrites. AB - A unique feature of the olfactory bulb circuit is the long projection of the mitral cell lateral dendrites. Through dendrodendritic reciprocal synapses, these dendrites connect one olfactory glomerular module to hundreds of others; but the functional principles governing these extensive lateral interactions remain largely unknown. Here we report that the spatial extent of action potential propagation in these dendrites is dynamically regulated by inhibitory synapses distributed along the dendrites. The extent of propagation determines the spatial pattern of Ca(2+) influx and thus the range and number of dendrodendritic synapses to be activated. Accordingly, network control of spike traffic in the mitral cell lateral dendrites can mediate dynamic interaction with different combinations of glomerular modules in response to different odorants. PMID- 11931747 TI - Tyrosine phosphatase STEP is a tonic brake on induction of long-term potentiation. AB - The functional roles of protein tyrosine phosphatases (PTPs) in the developed CNS have been enigmatic. Here we show that striatal enriched tyrosine phosphatase (STEP) is a component of the N-methyl-D-aspartate receptor (NMDAR) complex. Functionally, exogenous STEP depressed NMDAR single-channel activity in excised membrane patches. STEP also depressed NMDAR-mediated synaptic currents whereas inhibiting endogenous STEP enhanced these currents. In hippocampal slices, administering STEP into CA1 neurons did not affect basal glutamatergic transmission evoked by Schaffer collateral stimulation but prevented tetanus induced long-term potentiation (LTP). Conversely, inhibiting STEP in CA1 neurons enhanced transmission and occluded LTP induction through an NMDAR-, Src-, and Ca(2+)-dependent mechanism. Thus, STEP acts as a tonic brake on synaptic transmission by opposing Src-dependent upregulation of NMDARs. PMID- 11931748 TI - Perception of sound-source motion by the human brain. AB - We assessed the human brain network for sound-motion processing using the same virtual stimulus in three independent functional imaging experiments. All experiments show a bilateral posterior network of activation, including planum temporale (PT) and parieto-temporal operculum (PTO). This was demonstrated in contrasts between sound movement and two control conditions: externalized stationary stimuli (in the midline or to the side of the head) and midline sounds within the head with similar spectro-temporal structure. We suggest specific computational mechanisms in PT for disambiguation of the intrinsic spectro temporal features of a sound and the spectro-temporal effect of sound movement. The results support the existence of a posteriorly directed temporo-parietal pathway for obligatory perceptual processing of sound-source motion. PMID- 11931749 TI - Parallel visual motion processing streams for manipulable objects and human movements. AB - We tested the hypothesis that different regions of lateral temporal cortex are specialized for processing different types of visual motion by studying the cortical responses to moving gratings and to humans and manipulable objects (tools and utensils) that were either stationary or moving with natural or artificially generated motions. Segregated responses to human and tool stimuli were observed in both ventral and lateral regions of posterior temporal cortex. Relative to ventral cortex, lateral temporal cortex showed a larger response for moving compared with static humans and tools. Superior temporal cortex preferred human motion, and middle temporal gyrus preferred tool motion. A greater response was observed in STS to articulated compared with unarticulated human motion. Specificity for different types of complex motion (in combination with visual form) may be an organizing principle in lateral temporal cortex. PMID- 11931750 TI - BOLD activity during mental rotation and viewpoint-dependent object recognition. AB - We measured brain activity during mental rotation and object recognition with objects rotated around three different axes. Activity in the superior parietal lobe (SPL) increased proportionally to viewpoint disparity during mental rotation, but not during object recognition. In contrast, the fusiform gyrus was preferentially recruited in a viewpoint-dependent manner in recognition as compared to mental rotation. In addition, independent of the effect of viewpoint, object recognition was associated with ventral areas and mental rotation with dorsal areas. These results indicate that the similar behavioral effects of viewpoint obtained in these two tasks are based on different neural substrates. Such findings call into question the hypothesis that mental rotation is used to compensate for changes in viewpoint during object recognition. PMID- 11931751 TI - How does Pol II overcome the nucleosome barrier? AB - DNA packaging into chromatin presents a strong barrier to RNA polymerase II transcription. In this issue of Molecular Cell, Kireeva et al. describe a minimal system to examine polymerase II transcription through a positioned nucleosome and show, surprisingly, that transcription leads to the displacement of an H2A.H2B dimer from the nucleosome without altering nucleosome position. PMID- 11931752 TI - New structural clues to substrate specificity in the "ubiquitin system". AB - A 2.5 A crystal structure of a complex between the SUMO-conjugating enzyme Ubc9 and a protein substrate has yielded fresh insight into the specificity of protein modification by SUMO and other ubiquitin-like proteins. PMID- 11931753 TI - Structural surprises from the flaviviruses and alphaviruses. AB - Recent structural studies demonstrate that the alphavirus and flavivirus fusion proteins, although very similar in overall fold, are arranged very differently in the two virions. These differences raise many interesting questions about virus assembly and fusion activity. PMID- 11931754 TI - Half-pint: alternative splicing in the Drosophila ovary. AB - A new study from the Schupbach lab implicates a splicing factor, Half-pint, in the regulation of oogenesis in Drosophila. Through processing of the otu mRNA, Hfp appears to control both mitosis and RNA localization in the germline. PMID- 11931755 TI - Mechanisms of caspase activation and inhibition during apoptosis. AB - Caspases are central components of the machinery responsible for apoptosis. Recent structural and biochemical studies on procaspases, IAPs, Smac/DIABLO, and apoptosome have revealed a conserved mechanism of caspase activation and inhibition. This article reviews these latest advances and presents our current understanding of caspase regulation during apoptosis. PMID- 11931756 TI - Replication, recombination, and repair: going for the gold. AB - DNA recombination is now appreciated to be integral to DNA replication and cell survival. Recombination allows replication to successfully maneuver through the roadblocks of damaged or collapsed replication forks. The signals and controls that permit cells to transition between replication and recombination modes are now being identified. PMID- 11931757 TI - Human origin recognition complex large subunit is degraded by ubiquitin-mediated proteolysis after initiation of DNA replication. AB - Eukaryotic cells possess overlapping mechanisms to ensure that DNA replication is restricted to the S phase of the cell cycle. The levels of hOrc1p, the largest subunit of the human origin recognition complex, vary during the cell division cycle. In rapidly proliferating cells, hOrc1p is expressed and targeted to chromatin as cells exit mitosis and prereplicative complexes are formed. Later, as cyclin A accumulates and cells enter S phase, hOrc1p is ubiquitinated on chromatin and then degraded. hOrc1p destruction occurs through the proteasome and is signaled in part by the SCF(Skp2) ubiquitin-ligase complex. Other hORC subunits are stable throughout the cell cycle. The regulation of hOrc1p may be an important mechanism in maintaining the ploidy in human cells. PMID- 11931758 TI - Telomeric proteins regulate episomal maintenance of Epstein-Barr virus origin of plasmid replication. AB - Episomal maintenance and DNA replication of EBV origin of plasmid replication (OriP) plasmid maintenance is mediated by the viral encoded origin binding protein, EBNA1, and unknown cellular factors. We found that telomeric repeat binding factor 2 (TRF2), TRF2-interacting protein hRap1, and the telomere associated poly(ADP-ribose) polymerase (Tankyrase) bound to the dyad symmetry (DS) element of OriP in an EBNA1-dependent manner. TRF2 bound cooperatively with EBNA1 to the three nonamer sites (TTAGGGTTA), which resemble telomeric repeats. Mutagenesis of the nonamers reduced plasmid maintenance function and increased plasmid sensitivity to genotoxic stress. DS affinity-purified proteins possessed poly(ADP-ribose) polymerase (PARP) activity, and EBNA1 was subject to NAD dependent posttranslational modification in vitro. OriP plasmid maintenance was sensitive to changes in cellular PARP/Tankyrase activity. These findings imply that telomere-associated proteins regulate OriP plasmid maintenance by PAR dependent modifications. PMID- 11931759 TI - A positive-strand RNA virus replication complex parallels form and function of retrovirus capsids. AB - We show that brome mosaic virus (BMV) RNA replication protein 1a, 2a polymerase, and a cis-acting replication signal recapitulate the functions of Gag, Pol, and RNA packaging signals in conventional retrovirus and foamy virus cores. Prior to RNA replication, 1a forms spherules budding into the endoplasmic reticulum membrane, sequestering viral positive-strand RNA templates in a nuclease resistant, detergent-susceptible state. When expressed, 2a polymerase colocalizes in these spherules, which become the sites of viral RNA synthesis and retain negative-strand templates for positive-strand RNA synthesis. These results explain many features of replication by numerous positive strand RNA viruses and reveal that these viruses, reverse transcribing viruses, and dsRNA viruses share fundamental similarities in replication and may have common evolutionary origins. PMID- 11931760 TI - The dissociation of cohesin from chromosomes in prophase is regulated by Polo like kinase. AB - The separation of sister chromatids in anaphase depends on the dissociation of cohesin from chromosomes. In vertebrates, some cohesin is removed from chromosomes at the onset of anaphase by proteolytic cleavage. In contrast, the bulk of cohesin is removed from chromosomes already in prophase and prometaphase by an unknown mechanism that does not involve cohesin cleavage. We show that Polo like kinase is required for the cleavage-independent dissociation of cohesin from chromosomes in Xenopus. Cohesin phosphorylation depends on Polo-like kinase and reduces the ability of cohesin to bind to chromatin. These results suggest that Polo-like kinase regulates the dissociation of cohesin from chromosomes early in mitosis. PMID- 11931761 TI - Structure of the bacterial RNA polymerase promoter specificity sigma subunit. AB - The sigma subunit is the key regulator of bacterial transcription. Proteolysis of Thermus aquaticus sigma(A), which occurred in situ during crystallization, reveals three domains, sigma(2), sigma(3), and sigma(4), connected by flexible linkers. Crystal structures of each domain were determined, as well as of sigma(4) complexed with -35 element DNA. Exposed surfaces of each domain are important for RNA polymerase binding. Universally conserved residues important for -10 element recognition and melting lie on one face of sigma(2), while residues important for extended -10 recognition lie on sigma(3). Genetic studies correctly predicted that a helix-turn-helix motif in sigma(4) recognizes the -35 element but not the details of the protein-DNA interactions. Positive control mutants in sigma(4) cluster in two regions, positioned to interact with activators bound just upstream or downstream of the -35 element. PMID- 11931762 TI - Nucleosome remodeling induced by RNA polymerase II: loss of the H2A/H2B dimer during transcription. AB - RNA polymerase II (Pol II) must transcribe genes in a chromatin environment in vivo. We examined transcription by Pol II through nucleosome cores in vitro. At physiological and lower ionic strengths, a mononucleosome imposes a strong block to elongation, which is relieved at increased ionic strength. Passage of Pol II causes a quantitative loss of one H2A/H2B dimer but does not alter the location of the nucleosome. In contrast, bacteriophage SP6 RNA polymerase (RNAP) efficiently transcribes through the same nucleosome under physiological conditions, and the histone octamer is transferred behind SP6 RNAP. Thus, the mechanisms for transcription through the nucleosome by Pol II and SP6 RNAP are clearly different. Moreover, Pol II leaves behind an imprint of disrupted chromatin structure. PMID- 11931763 TI - Nuclear receptor function requires a TFTC-type histone acetyl transferase complex. AB - Nuclear receptors (NRs) regulate transcription in a ligand-dependent way through two types of coactivator complexes: the p160/CBP histone acetyl transferase (HAT) complex and the DRIP/TRAP/SMCC complex without HAT activity. Here we identified a large human (h) coactivator complex necessary for the estrogen receptor alpha (ERalpha) transactivation. This complex contains the GCN5 HAT, the c-Myc interacting protein TRRAP/PAF400, TAF(II)30, and other subunits. Similarly to known TFTC (TBP-free TAF(II)-containing)-type HAT complexes (hTFTC, hPCAF, and hSTAGA), TRRP directly interacted with liganded ER alpha, or other NRs. ER alpha transactivation was enhanced by the purified complex in vitro. Antisense TRRAP RNA inhibited estrogen-dependent cell growth of breast cancer cells. Thus, the isolated TFTC-type HAT complex acts as a third class of coactivator complex for NR function. PMID- 11931764 TI - The genome-wide localization of Rsc9, a component of the RSC chromatin-remodeling complex, changes in response to stress. AB - The cellular response to environmental changes includes widespread modifications in gene expression. Here we report the identification and characterization of Rsc9, a member of the RSC chromatin-remodeling complex in yeast. The genome-wide localization of Rsc9 indicated a relationship between genes targeted by Rsc9 and genes regulated by stress; treatment with hydrogen peroxide or rapamycin, which inhibits TOR signaling, resulted in genome-wide changes in Rsc9 occupancy. We further show that Rsc9 is involved in both repression and activation of mRNAs regulated by TOR as well as the synthesis of rRNA. Our results illustrate the response of a chromatin-remodeling factor to signaling cascades and suggest that changes in the activity of chromatin-remodeling factors are reflected in changes in their localization in the genome. PMID- 11931765 TI - Structural basis of lysine-acetylated HIV-1 Tat recognition by PCAF bromodomain. AB - The human immunodeficiency virus type 1 (HIV-1) trans-activator protein Tat stimulates transcription of the integrated HIV-1 genome and promotes viral replication in infected cells. Tat transactivation activity is dependent on lysine acetylation and its association with nuclear histone acetyltransferases p300/CBP (CREB binding protein) and p300/CBP-associated factor (PCAF). Here, we show that the bromodomain of PCAF binds specifically to HIV-1 Tat acetylated at lysine 50 and that this interaction competes effectively against HIV-1 TAR RNA binding to the lysine-acetylated Tat. The three-dimensional solution structure of the PCAF bromodomain in complex with a lysine 50-acetylated Tat peptide together with biochemical analyses provides the structural basis for the specificity of this molecular recognition and reveals insights into the differences in ligand selectivity of bromodomains. PMID- 11931766 TI - Promoter-specific regulation of MyoD binding and signal transduction cooperate to pattern gene expression. AB - We used expression arrays and chromatin immunoprecipitation assays to demonstrate that myogenesis consists of discrete subprograms of gene expression regulated by MyoD. Approximately 5% of assayed genes alter expression in a specific temporal sequence, and more than 1% are regulated by MyoD without the synthesis of additional transcription factors. MyoD regulates genes expressed at different times during myogenesis, and promoter-specific regulation of MyoD binding is a major mechanism of patterning gene expression. In addition, p38 kinase activity is necessary for the expression of a restricted subset of genes regulated by MyoD, but not for MyoD binding. The identification of distinct molecular mechanisms that regulate discrete subprograms of myogenesis should facilitate analyses of differentiation in normal development and disease. PMID- 11931767 TI - Formation of the androgen receptor transcription complex. AB - Androgen receptor (AR) is required for sexual differentiation and is implicated in the development of prostate cancer. Here we describe distinct functions for cofactor proteins and gene regulatory elements in the assembly of AR-mediated transcription complexes. The formation of an activation complex involves AR, coactivators, and RNA polymerase II recruitment to both the enhancer and promoter, whereas the formation of a repression complex involves factors bound only at the promoter and not the enhancer. These results suggest a model for the functional coordination between the promoter and enhancer in which communication between these elements is established through shared coactivators in the AR transcription complex. PMID- 11931768 TI - The N-CoR-HDAC3 nuclear receptor corepressor complex inhibits the JNK pathway through the integral subunit GPS2. AB - The corepressors N-CoR and SMRT partner with histone deacetylases (HDACs) in diverse repression pathways. We report here that GPS2, a protein involved in intracellular signaling, is an integral subunit of the N-CoR-HDAC3 complex. We have determined structural motifs that direct the formation of a highly stable and active deacetylase complex. GPS2 and TBL1, another component of the N-CoR HDAC3 complex, interact cooperatively with repression domain 1 of N-CoR to form a heterotrimeric structure and are indirectly linked to HDAC3 via an extended N-CoR SANT domain that also activates latent HDAC3 activity. More importantly, we show here that the N-CoR-HDAC3 complex inhibits JNK activation through the associated GPS2 subunit and thus could potentially provide an alternative mechanism for hormone-mediated antagonism of AP-1 function. PMID- 11931769 TI - The phosphorylation status of nuclear NF-kappa B determines its association with CBP/p300 or HDAC-1. AB - Homodimers of the NF-kappa B p50 subunit are transcriptionally repressive in cells, whereas they can promote transcription in vitro, suggesting that their endogenous effects are mediated by association with other factors. We now demonstrate that transcriptionally inactive nuclear NF-kappaB in resting cells consists of homodimers of either p65 or p50 complexed with the histone deacetylase HDAC-1. Only the p50-HDAC-1 complexes bind to DNA and suppress NF kappa B-dependent gene expression in unstimulated cells. Appropriate stimulation causes nuclear localization of NF-kappa B complexes containing phosphorylated p65 that associates with CBP and displaces the p50-HDAC-1 complexes. Our results demonstrate that phosphorylation of p65 determines whether it associates with either CBP or HDAC-1, ensuring that only p65 entering the nucleus from cytoplasmic NF-kappa B:Ikappa B complexes can activate transcription. PMID- 11931770 TI - Peptide-induced negative selection of thymocytes activates transcription of an NF kappa B inhibitor. AB - Negative selection eliminates thymocytes bearing autoreactive T cell receptors (TCR) via an apoptotic mechanism. We have cloned an inhibitor of NF-kappa B, I kappa BNS, which is rapidly expressed upon TCR-triggered but not dexamethasone- or gamma irradiation-stimulated thymocyte death. The predicted protein contains seven ankyrin repeats and is homologous to I kappa B family members. In class I and class II MHC-restricted TCR transgenic mice, transcription of I kappa BNS is stimulated by peptides that trigger negative selection but not by those inducing positive selection (i.e., survival) or nonselecting peptides. I kappa BNS blocks transcription from NF-kappa B reporters, alters NF-kappa B electrophoretic mobility shifts, and interacts with NF-kappa B proteins in thymic nuclear lysates following TCR stimulation. Retroviral transduction of I kappa BNS in fetal thymic organ culture enhances TCR-triggered cell death consistent with its function in selection. PMID- 11931771 TI - Dynamic antagonism between ETR-3 and PTB regulates cell type-specific alternative splicing. AB - Inclusion of cardiac troponin T (cTNT) exon 5 in embryonic muscle requires conserved flanking intronic elements (MSEs). ETR-3, a member of the CELF family, binds U/G motifs in two MSEs and directly activates exon inclusion in vitro. Binding and activation by ETR-3 are directly antagonized by polypyrimidine tract binding protein (PTB). We use dominant-negative mutants to demonstrate that endogenous CELF and PTB activities are required for MSE-dependent activation and repression in muscle and nonmuscle cells, respectively. Combined use of CELF and PTB dominant-negative mutants provides an in vivo demonstration that antagonistic splicing activities exist within the same cells. We conclude that cell-specific regulation results from the dominance of one among actively competing regulatory states rather than modulation of a nonregulated default state. PMID- 11931772 TI - Identity elements used in export of mRNAs. AB - Different classes of RNA are exported from the nucleus by distinct factors. We demonstrate that U1 snRNA is exported like an mRNA on insertion of a pre-mRNA intron or either sense or antisense mRNA exon sequences. mRNA-specific factors are recruited onto the spliced or elongated U1 RNA whereas U snRNA-specific factors are not, suggesting that an unstructured region of sufficient length in an RNA acts as a dominant determinant of mRNA identity. After export, spliced U1 RNA undergoes cytoplasmic maturation but is not reimported into the nucleus. These data provide insight into mechanisms for discrimination of different classes of nuclear RNA and demonstrate that two RNAs of identical sequence can have distinct cytoplasmic fates depending on their mode of export. PMID- 11931773 TI - ClpS, a substrate modulator of the ClpAP machine. AB - In the bacterial cytosol, ATP-dependent protein degradation is performed by several different chaperone-protease pairs, including ClpAP. The mechanism by which these machines specifically recognize substrates remains unclear. Here, we report the identification of a ClpA cofactor from Escherichia coli, ClpS, which directly influences the ClpAP machine by binding to the N-terminal domain of the chaperone ClpA. The degradation of ClpAP substrates, both SsrA-tagged proteins and ClpA itself, is specifically inhibited by ClpS. In contrast, ClpS enhanced ClpA recognition of two heat-aggregated proteins in vitro and, consequently, the ClpAP-mediated disaggregation and degradation of these substrates. We conclude that ClpS modifies ClpA substrate specificity, potentially redirecting degradation by ClpAP toward aggregated proteins. PMID- 11931774 TI - Structural basis and specificity of acyl-homoserine lactone signal production in bacterial quorum sensing. AB - Synthesis and detection of acyl-homoserine lactones (AHLs) enables many gram negative bacteria to engage in quorum sensing, an intercellular signaling mechanism that activates differentiation to virulent and biofilm lifestyles. The AHL synthases catalyze acylation of S-adenosyl-L-methionine by acyl-acyl carrier protein and lactonization of the methionine moiety to give AHLs. The crystal structure of the AHL synthase, EsaI, determined at 1.8 A resolution, reveals a remarkable structural similarity to the N-acetyltransferases and defines a common phosphopantetheine binding fold as the catalytic core. Critical residues responsible for catalysis and acyl chain specificity have been identified from a modeled substrate complex and verified through functional analysis in vivo. A mechanism for the N-acylation of S-adenosyl-L-methionine by 3-oxo-hexanoyl-acyl carrier protein is proposed. PMID- 11931775 TI - Immunologic graft reactions after allogenic penetrating keratoplasty. AB - PURPOSE: To evaluate frequency and risk factors of immunologic graft reactions after allogenic penetrating keratoplasty. DESIGN: Interventional comparative nonrandomized clinical trial. METHODS: The setting took place in a university eye hospital. The retrospective study included 338 patients (338 eyes). The patients underwent penetrating keratoplasty performed by a single surgeon in the study period from 1989 to 1997. Follow-up period had to be longer than 12 months (mean +/- SD, 31.4 +/- 18.8 months). Frequency of immunologic graft reactions characterized by relatively few small monomorph whitish cells in the anterior chamber, almost no flare, and retrocorneal cellular precipitates. RESULTS: Immunologic graft reactions were detected in 46 patients (46/338 = 13.6%). Statistically significant risk factors for the development of graft reactions were loosening of sutures (P =.046), and preoperative and postoperative corneal vascularization (P =.04). Frequency of an immunologic graft reaction was statistically independent (P >.05) of the graft diameters used in the present study, age, and gender of the patients, HLA-typing, donor age, and preservation data of the donor material. Seventy-four percent (34/46) of all graft reactions were detected within the first 2.5 years after surgery. Thirteen percent (6/46) of all graft reactions were observed more than 4 years after keratoplasty. With intensive corticosteroid treatment, graft transparency could be regained in 44 (95.6%) of the 46 patients with an immunologic graft reaction. CONCLUSIONS: Most important risk factors for immunologic graft reactions occurring in approximately 14% of patients after allogenic penetrating keratoplasty are suture loosening and preoperative and postoperative corneal vascularization. Graft diameters as used in the present study, HLA-typing, age of the donor, and preservation data of the donor material may not play a major role. More than 10% of graft reaction episodes can occur more than 4 years postgrafting. With intensive corticosteroid treatment, graft transparency can be regained in the majority of patients after an immunologic graft reaction when detected early. PMID- 11931776 TI - Noncontact corneal pachymetry with slit lamp-adapted optical coherence tomography. AB - PURPOSE: The purpose of this study was to determine the accuracy, the reproducibility, and the limits of agreement of noncontact central corneal thickness measurement with slit lamp-adapted optical coherence tomography (OCT). DESIGN: Nonrandomized comparative clinical trial. METHODS: In a prospective comparative observational study, a total of 108 consecutive patients (108 eyes) with normal corneas (92 eyes) and different corneal alterations (16 eyes) participated. Six sequential measurements of the central corneal thickness with slit lamp-adapted OCT and with ultrasound (US) pachymetry at 1640 ms(-1) were performed. The main outcome measures were accuracy, reproducibility assessed with precision and coefficient of variation (CV), and limits of agreement of central corneal thickness measurement. RESULTS: The mean central corneal thickness values were 541 +/- 43 microm (OCT) and 549 +/- 44 microm (US) with a mean precision of +/- 5.8 microm (CV 1.08%) and of +/- 4.0 microm (CV 0.73%), respectively. The method comparison revealed equivalence (+/- 2SD) in the 5% range with a mean difference between both methods of 7.9 microm (1.45%). The relative error was 8.7 microm (1.6%), which corresponded to limits of agreement (+/- 2SD) ranging from 9.5 microm to 25.3 microm. CONCLUSIONS: Central corneal pachymetry with slit lamp adapted OCT revealed, for clinical purposes, an excellent accuracy and reproducibility with a high degree of agreement compared with US pachymetry. Thus, the presented OCT system seems to be a promising diagnostic modality to objectively measure corneal thickness in a convenient noncontact mode. PMID- 11931777 TI - Effective treatment of ligneous conjunctivitis with topical plasminogen. AB - PURPOSE: The etiology of ligneous conjunctivitis is now known to be due to an underlying type 1 plasminogen deficiency. We hereby report the clinical features of three cases and their response to topically administered plasminogen. DESIGN: Observational case series. METHODS: Two Caucasian females aged 5 years and an 18 month male of north African descent presented with a membranous conjunctivitis, which recurred after surgical excision. Case 1 presented before the association with plasminogen deficiency was known with a bilateral chronic membranous mucopurulent conjunctivitis from the age of 14 months associated with bronchiolitis and gingival hyperplasia. A diagnosis of ligneous conjunctivitis was entertained and a number of drops were instituted. At the age of 4 years plasminogen levels were ordered. Case 2 presented at the age of 4 years with a unilateral chronic membranous conjunctivitis. Plasminogen levels were requested as soon as a diagnosis of ligneous conjunctivitis was suspected. Case 3 was born with congenital hydrocephalus. Conjunctivitis was treated with antibiotics from the age of 1 month. He presented to the eye clinic at the age of 5 months when a clinical diagnosis of ligneous conjunctivitis was entertained and treated with a number of medications. Plasminogen levels were available at 9 months of age. RESULTS: The two female patients returned plasminogen levels of 0.25 U/ml and 0.3 U/ml, well below the normal level of 0.7-1.0 U/ml. Functional plasminogen levels in the male infant were not recordable with plasminogen antigen levels of 0.125 U/ml (normal range, 0.52-1.82). All cases have responded well to excision of the membranes and institution of topical plasminogen drops. There has been no recurrence with more than 12 months' follow-up. CONCLUSIONS: With the knowledge of the etiology of ligneous conjunctivitis, efforts are underway to identify the best method of delivery of plasminogen. Topical plasminogen concentrate from fresh frozen plasma holds promise as the definitive treatment for this chronic membranous conjunctivitis PMID- 11931778 TI - Late-onset transconjunctival oozing and point leak of aqueous humor from filtering bleb after trabeculectomy. AB - PURPOSE: To estimate the prevalence and identify factors related to late-onset transconjunctival aqueous oozing and point leak from functioning blebs after trabeculectomy with 5-fluorouracil (5-FU) or mitomycin C. DESIGN: Cross-sectional study. METHODS: SETTING: The study took place at the outpatient clinic of the Department of Ophthalmology, University of Tokyo Graduate School of Medicine (Tokyo, Japan). Four hundred three consecutive patients (403 eyes) with functional blebs at least 3 months after previously performed trabeculectomy were examined between December 1997 and February 1998. The Seidel test was performed with extended observation up to 15 seconds. Oozing was identified as transconjunctival aqueous egress without interruption of the conjunctival tissue or aqueous stream on the bleb wall. Logistic regression analyses of oozing and point leak in 331 eyes with an avascular area were performed using independent variables, including age, gender, glaucoma diagnosis, prior incisional surgery, antimetabolite use, combined cataract surgery, postoperative follow-up period, intraocular pressure (IOP), concurrent glaucoma therapy, bleb size, and avascular area size. RESULTS: Of 403 eyes, 48 eyes (11.9%) had oozing and 8 eyes (2.0%) had point leak. Intraocular pressure was significantly lower and an avascular area was more frequent in eyes with oozing or leak than in eyes without (P <.001). Logistic regression analysis revealed that oozing was significantly more common after use of 5-FU than mitomycin C (P =.024), whereas point leak was associated with a larger avascular area (P =.045). CONCLUSIONS: After trabeculectomy with antimetabolites, transconjunctival oozing is much more frequent than point leak. Oozing was significantly associated with the use of 5-FU and point leak with a larger avascular area in the bleb. PMID- 11931779 TI - Fourth generation fluoroquinolones: new weapons in the arsenal of ophthalmic antibiotics. AB - PURPOSE: Fourth generation fluoroquinolones (FQs) will soon be introduced to ophthalmology. In this in vitro study, differences in the susceptibility patterns and the potencies of fourth generation FQs (gatifloxacin-GAT and moxifloxacin MOX) were compared with third generation (levofloxacin-LEV) and second generation FQs (ciprofloxacin-CIP and ofloxacin-OFX). DESIGN: Experimental laboratory investigation. METHODS: In retrospect, the minimum inhibitory concentrations (MICs) of 93 bacterial endophthalmitis isolates were determined to CIP, OFX, LEV, GAT, and MOX using E-tests. The National Committee of Clinical Laboratory Standards (NCCLS) susceptibility patterns and the potencies of the MICs were statistically compared. RESULTS: With in vitro tests, Staphylococcus aureus isolates that were resistant to CIP and OFX were statistically most susceptible (P =.01) to MOX. Coagulase negative Staphylococci that were resistant to CIP and OFX were statistically most susceptible (P =.02) to MOX and GAT. Streptococcus viridans were more susceptible (P =.02) to MOX, GAT, and LEV than CIP and OFX. Streptococcus pneumoniae was least susceptible (P =.01) to OFX compared with the other FQs. Susceptibilities were equivalent (P =.11) for all other bacterial groups. In general, MOX was the most potent FQ for gram-positive bacteria (P =.05) while CIP, MOX, GAT, and LEV demonstrated equivalent potencies to gram negative bacteria. CONCLUSIONS: This in vitro study indicated that fourth generation FQs appear to cover bacterial resistance to the second and third generation FQs, were more potent than the second and third generation FQs for gram-positive bacteria, and are equally potent for gram-negative bacteria. Clinical studies will need to confirm these results. PMID- 11931780 TI - A randomized controlled clinical trial of intravitreous fomivirsen for treatment of newly diagnosed peripheral cytomegalovirus retinitis in patients with AIDS. AB - PURPOSE: To assess the efficacy of intravitreous fomivirsen sodium, an antisense oligonucleotide, for newly diagnosed peripheral cytomegalovirus (CMV) retinitis in patients with acquired immunodeficiency syndrome (AIDS). DESIGN: Randomized treatment intervention clinical trial. METHODS: A multicenter, prospective, and randomized clinical trial compared immediate treatment of CMV retinitis with fomivirsen (165 microg administered intravitreously) to deferral of treatment until CMV retinitis lesions progressed by standard definitions. Included were patients with CMV retinitis lesions at least 750 microm outside of zone 1. Fomivirsen was injected weekly for three doses as induction therapy, followed by injection every other week as maintenance therapy. All patients were examined regularly until evidence of retinitis progression. Time to first progression was determined by two independent masked fundus photography reading centers (principal outcome) and by clinician investigators based on indirect ophthalmoscopy. Patients in the deferral of treatment group were offered fomivirsen therapy at the time of clinically determined retinitis progression. RESULTS: Patients in the immediate treatment group (n = 18) and the deferral of treatment group (n = 10) were comparable at baseline. Median time to first progression of disease for the immediate treatment group was 71 days (95% confidence interval [CI] 28 days-not determinable) and for the deferral of treatment group was 13 days (95% CI 9-15 days; P =.0001, Wilcoxon rank sum test). Progression occurred in 44% of patients in the immediate treatment group during the study compared with 70% of patients in the deferral of treatment group during the study. There were no retinal detachments among eyes treated with fomivirsen. CONCLUSIONS: Fomivirsen is an effective treatment for CMV retinitis in patients with AIDS that utilizes a mechanism of action different than that of ganciclovir, foscarnet, and cidofovir. PMID- 11931781 TI - Randomized dose-comparison studies of intravitreous fomivirsen for treatment of cytomegalovirus retinitis that has reactivated or is persistently active despite other therapies in patients with AIDS. AB - PURPOSE: To compare the efficacy of two regimens of intravitreous fomivirsen for treatment of cytomegalovirus (CMV) retinitis that had reactivated or was persistently active despite other anti-CMV therapies in patients with acquired immunodeficiency syndrome (AIDS). DESIGN: Randomized treatment intervention clinical trials. METHODS: Two multicenter prospective randomized open-label controlled clinical trials (USA/Brazilian study; EuroCanadian study) each compared two treatment regimens of intravitreous fomivirsen (330 microg/injection), a more intense schedule (regimen A) consisting of 3 weekly injections as induction, followed by injections every 2 weeks as maintenance therapy, and a less intense schedule (regimen B), consisting of two injections at day 1 and day 15 as induction, followed by injections every 4 weeks as maintenance therapy. A total of 61 patients (67 eyes) were treated with regimen A and a total of 32 patients (39 eyes) were treated with regimen B. Time to progression was determined by masked evaluation of serial fundus photographs. The proportion of patients whose lesions became less active or inactive is described for the USA/Brazilian study as an additional measure of efficacy. RESULTS: Results were similar for both regimens. In the USA/Brazilian study, the median time to progression was 106 days for regimen A and 267 days for regimen B (P =.2179, Wilcoxon rank sum test). In the EuroCanadian study, the median time to progression was not determinable for regimen A (25th percentile: 91 days), but was 403 days for regimen B. In the USA/Brazilian study, lesion activity decreased in 80% of patients and became inactive in 55% of patients during therapy. CONCLUSIONS: Both regimens of intravitreous fomivirsen can be effective treatment for patients with CMV retinitis that has not been controlled by other anti-CMV drugs. The less intense regimen may be preferable in terms of convenience and safety. PMID- 11931782 TI - Safety of intravitreous fomivirsen for treatment of cytomegalovirus retinitis in patients with AIDS. AB - PURPOSE: To report data regarding the safety of intravitreous fomivirsen for treatment of cytomegalovirus (CMV) retinitis in patients with acquired immunodeficiency syndrome (AIDS). DESIGN: Critical review of safety data from three randomized controlled clinical trials with supplemental information from an expanded drug access program. METHODS: Adverse ocular events reported by clinician investigators were listed using terms modified from the COSTART dictionary. Data for two doses (165-microg/injection [35 eyes, 30 patients] and 330-microg/injection [153 eyes, 120 patients]) and two 330-microg/injection dose schedules of different intensity were pooled to calculate incidence rates for each event. Rates were calculated as "events/patient-year" (based on total cumulative reported events and duration of treatment) for events that could recur during treatment. Rates were calculated as "patients with events/person-year" for the following events: retinal detachment, cataract, visual field disturbance, and retinal pigment epitheliopathy. To assess the ability to manage events, we reviewed treatments given for two events (anterior chamber inflammation, increased intraocular pressure) in one trial. We also report an analysis comparing the proportion of eyes that developed one or more key events to the cumulative number of injections. RESULTS: Incidence rates were dose and schedule dependent (165 microg/injection, 4.06 events/patient-year; 330 microg/injection, 6.58 events/patient-year [less intense regimen] and 8.35 events/patient-year [more intense regimen]). The most frequently reported events were anterior chamber inflammation and increased intraocular pressure. We found no evidence that the proportion of patients with events increased as the number of injections increased. CONCLUSIONS: Intravitreous fomivirsen is well tolerated with an acceptable safety profile. Adverse ocular events associated with doses and schedules used clinically can be managed successfully with medical therapy. PMID- 11931783 TI - Chest computerized tomography in the evaluation of uveitis in elderly women. AB - PURPOSE: To describe the usefulness of chest computerized tomography (CT) in the evaluation of uveitis in elderly women and the clinical characteristics of patients with an abnormal chest CT scan. DESIGN: Prospective noncomparative case series. METHODS: We evaluated 30 elderly women, aged 61-83 years, with chronic iritis, vitritis, or choroiditis and with no definitive cause for their uveitis. All patients underwent a battery of diagnostic laboratory studies and chest CT. RESULTS: The diagnostic examination in most patients included serum angiotensin converting enzyme level, serum lysozyme, rapid plasma reagin level, fluorescent treponemal antibody-absorption test, purified protein derivative skin test, and chest x-rays. Chest CT performed on all patients showed parenchymal, mediastinal, and/or hilar adenopathy in 17 patients (57%). Histopathologic confirmation of sarcoidosis with noncaseating granulomas in the biopsy specimens was obtained in 14 patients: eight by mediastinoscopy, two by bronchoscopy, two by conjunctival biopsy, one by nasal biopsy, and one by vitreous biopsy. CONCLUSIONS: Chest CT can be useful in elderly female patients with chronic uveitis for identifying mediastinal lymphadenopathy and other lesions suggestive of sarcoidosis, as well as to help guide tissue confirmation and to rule out other diagnoses including lymphoma. PMID- 11931784 TI - Ocular toxoplasmosis after the fifth decade. AB - PURPOSE: To describe the clinical features in patients presenting with ocular toxoplasmosis after the fifth decade and to analyze laboratory findings in comparison to uveitis history and clinical data. DESIGN: Prospective consecutive observational case series. METHODS: A prospective clinical analysis of 27 consecutive patients older than 50 years of age with primary or recurrent ocular toxoplasmosis was performed during a period of 8 years. These cases account for 12% of all ocular toxoplasmosis cases irrespective of age indexed in our institution during the same period. Paired serum and aqueous humor samples were tested for anti-Toxoplasma gondii IgG, IgM, and IgA antibodies. The presence of T. gondii DNA in aqueous humor was determined by polymerase chain reaction followed by DNA hydridization method. RESULTS: Although similar in age, two groups were distinguished clinically: 12 patients (44%) presented with usual forms of retinochoroiditis (mean +/- SD, 1.6 +/- 0.5 disk areas [DA] in size); 15 patients (56%) presented with atypical lesions, greater than 3 DA in size (mean +/- SD, 5.0 +/- 2.0 DA). The second group showed a higher rate of complications (P =.028) and a poorer visual outcome (P =.015). Twenty-four patients (89%) had intraocular IgG production, 17 (63%) had intraocular IgA production, 3 (11%) had intraocular IgM production, and 12 (44%) had a positive T. gondii DNA detection. CONCLUSIONS: After the fifth decade, ocular toxoplasmosis remains an important cause of posterior uveitis. The combination of antibody detection by immunocapture tests with T. gondii DNA detection, both in aqueous humor, allowed the diagnosis of toxoplasmic infection in the atypical cases with large ocular lesions. PMID- 11931785 TI - Persistent foveal retinal detachment after successful rhegmatogenous retinal detachment surgery. AB - PURPOSE: To describe the tomographic features of the neurosensory retina after successful rhegmatogenous retinal detachment surgery. DESIGN: Cohort study. METHODS: We prospectively examined cross-sectional retinal images using optical coherence tomography (OCT) in 15 eyes of 15 consecutive patients (mean age, 46 years) who underwent scleral buckling surgery for treatment of rhegmatogenous retinal detachment. The retinas appeared reattached upon binocular stereoscopic indirect ophthalmoscopy. All eyes were examined at 1, 3, 6, and 12 months postoperatively. The time from onset of subjective symptoms of retinal detachment to retinal surgery ranged from 4 to 66 days (mean, 14 days). The postoperative OCT findings and best-corrected visual acuity were statistically analyzed using the Mann-Whitney U test. RESULTS: The detached retinas appeared attached on ophthalmoscopy in all eyes 1 month postoperatively. OCT showed residual retinal detachment in four eyes (27%) at the fovea and in three eyes (20%) at the fovea and adjacent area. The remaining eight retinas (53%) were attached when examined tomographically. In six of the seven eyes with residual foveal detachment by OCT, the retinas reattached spontaneously up to 12 months postoperatively. The postoperative best-corrected visual acuity improved gradually during 12 postoperative months in all 15 eyes. A substantial increase in visual acuity occurred when the fovea reattached in the six eyes with residual detachment. CONCLUSIONS: Foveal retinal detachment may persist after successful retinal detachment surgery in eyes in which the fovea appeared to be attached on ophthalmoscopy. The residual foveal detachment may explain, in part, the delayed visual acuity improvement after successful scleral buckling. PMID- 11931786 TI - Radiotherapy for subfoveal choroidal neovascularization in age-related macular degeneration: a randomized clinical trial. AB - PURPOSE: To report results of 18-month follow up of external beam radiation therapy with photons for subfoveal classic or occult choroidal neovascularization (CNV) in age-related macular degeneration (ARMD). DESIGN: Randomized clinical trial. METHODS: A total of 161 patients with subfoveal CNV in ARMD were recruited in a prospective double-masked study. The posterior pole of the afflicted eye was given 1 Gy (4 x 0.25 Gy) in the control group and 8 Gy (4 x 2 Gy) or 16 Gy (4 x 4 Gy) in the treatment groups. At the time of treatment, and 6, 12, and 18 months post treatment, best-corrected visual acuity (BCVA), reading ability, and CNV size were measured. RESULTS: At the completion of the study 150 (93.2%), 139 (86.3%), and 137 (85.1%) patients were followed for 6, 12, and 18 months, respectively. The mean number of lines lost in the BCVA was -1.69, -2.2, and 3.23 in the 1 Gy group; -0.94, -1.25, and -1.73 in the 8 Gy group; -0.51, -0.67, and -1.93 in the 16 Gy group. The difference was significant after 12 months (P =.016 for 8 Gy vs. 1 Gy; P =.006 for 16 Gy vs. 1 Gy), and 18 months (P =.011 for 8 Gy vs. 1 Gy; P =.05 for 16 Gy vs. 1 Gy). The patients with classic CNV, or with an initial distance visual acuity >or=20/100, benefited more from treatment. A significant difference was not found between control group and treatment groups in the reading ability and in the CNV size. No radiation-associated side effects were reported thus far. CONCLUSION: The number of lines lost in the BCVA was less in the 8 Gy and 16 Gy treatment groups than in the control group during the complete follow up examination. Radiation therapy with 8 Gy and 16 Gy, without showing any difference in efficacy, resulted in a near stabilization of the BCVA in patients with subfoveal classic or occult CNV in ARMD. Further studies are necessary to determine the significance of repeated radiotherapy series with a dose of 8 Gy to improve the effect on the CNV size and thereby to prolong stabilization of distance visual acuity. PMID- 11931787 TI - Surgical treatment of subfoveal neovascularization in myopia: macular translocation vs surgical removal. AB - PURPOSE: To compare the visual outcome of two different surgical approaches for subfoveal neovascularization in degenerative myopia: macular translocation and surgical removal of choroidal neovascularization (CNV). DESIGN: Interventional case series. METHODS: Retrospectively, 32 eyes with degenerative myopia (axial length over 26 mm or refraction over -6 diopters) and subfoveal CNV of 32 consecutive patients operated on by either surgical removal of CNV or limited macular translocation were reviewed. Surgical removal of CNV was performed in 18 eyes and limited macular translocation with a twofold suture in 14 eyes. The main outcome measurements were best-corrected visual acuity (BCVA) and findings from fluorescein angiography. Postoperatively, mean +/- SD follow up was 14 +/- 15 months (range, 6-48 months) in the removal group and 11 +/- 4 months (range, 6-24 months, P =.37) in the translocation group. RESULTS: In both groups, there was no significant difference in preoperative age, sex, refractive error, or BCVA. The average of postoperative BCVA was statistically better after macular translocation (10 ETDRS lines or 20/100) than after surgical removal (6 lines 10/125, P =.019). Visual acuity improved by 3.8 lines after macular translocation and was unchanged after surgical removal (-0.7 line, P =.011). Macular translocation was successful in shifting the CNV to an extrafoveal location in 11 out of 14 eyes. Mean foveal displacement of all 18 translocated eyes was 695 +/- 426 microm (range, 100-1520 microm). Recurrence of CNV occurred in seven eyes (39%) after surgical removal and in two eyes (14%) after translocation. Retinal detachment occurred in two eyes in each group. CONCLUSION: In this retrospective study, eyes with degenerative myopia and subfoveal neovascularization treated with limited macular translocation had better visual acuity recovery than eyes treated with surgical removal of the choroidal neovascularization. Further studies are required to confirm these results. PMID- 11931789 TI - Transplantation of intact sheets of fetal neural retina with its retinal pigment epithelium in retinitis pigmentosa patients. AB - PURPOSE: To show the safety of transplanting sheets of fetal neural retina together with its retinal pigment epithelium (RPE) to patients with retinitis pigmentosa. DESIGN: Interventional case series. METHODS: Sheets of fetal neural retina and RPE were transplanted together into the subretinal space near the fovea unilaterally in the eyes of five patients with retinitis pigmentosa who had only light perception in both eyes. The patients were followed for 6 months. The main outcome measures were tissue typing of both donors and recipients, fluorescein angiography, multifocal electroretinogram (mfERG) testing, and clinical examination. No immunosuppressive medications were given. RESULTS: No evidence of rejection was observed. Up to 6 months there was no evidence of tissue disintegration, retinal edema, or scarring. There was no change in vision both by Snellen acuity and with mfERGs. Growth of the transplant was noted in two of five patients at 6 months vs. 2 weeks. All patients typed were HLA mismatched with donor tissue. CONCLUSIONS: This study indicates that fetal retina can be transplanted together with its RPE and survive for at least 6 months without evidence of rejection. However, no improvements in vision were observed, possibly due to the severe retinal degeneration of the patients. PMID- 11931788 TI - Angiotensin II and vascular endothelial growth factor in the vitreous fluid of patients with diabetic macular edema and other retinal disorders. AB - PURPOSE: To investigate the correlation between angiotensin II (AII) or vascular endothelial growth factor (VEGF) levels in the vitreous fluid and the severity of diabetic macular edema (DME). DESIGN: A case-control study. METHODS: Vitreous fluid samples were obtained at the time of vitreoretinal surgery from 20 eyes of 20 patients with DME, 6 eyes of 6 diabetic patients without retinopathy, and 14 eyes of 14 nondiabetic patients. The VEGF levels in vitreous fluid and plasma were determined by enzyme-linked immunosorbent assay, while AII levels were measured by radioimmunoassay. RESULTS: The vitreous concentration of VEGF was significantly higher in patients with DME than in nondiabetic patients or diabetic patients without retinopathy (P <.0001 and P <.0001, respectively). Vitreous levels of AII were also higher in patients with DME than in nondiabetic patients (P =.0082). The vitreous concentration of AII was significantly correlated with that of VEGF (P =.0022). Vitreous concentrations of both AII and VEGF were significantly higher in patients with hyperfluorescent DME than in those with hypofluorescent (P =.0228 and P =.0068, respectively). CONCLUSIONS: We found that the levels of both AII and VEGF were elevated in the vitreous fluid of patients with hyperfluorescein DME. Our results suggest that both AII and VEGF are related to the increase of vascular permeability in DME. PMID- 11931790 TI - InternetAdvance: accelerated internet publication of articles and reports at ajo.com. PMID- 11931791 TI - Fomivirsen for the treatment of cytomegalovirus retinitis. PMID- 11931792 TI - The association of HLA with young-onset keratoconus in Japan. AB - PURPOSE: To report the association of HLA antigens with keratoconus in Japanese patients. DESIGN: Observational consecutive case series. METHODS: In 90 consecutive Japanese keratoconus patients, HLA class I (HLA-A, -B, -C) and class II (HLA-DR, -DQ) were analyzed. RESULTS: Compared with control frequencies, based on mean gene frequencies for the Japanese population, higher frequencies of HLA A26, B40, and DR9 antigens were found in patients whose conditions were diagnosed before 20 years of age (chi(2) = 6.45, P =.01; chi(2) = 6.78, P =.01; chi(2) =3.99, P =.05, respectively), but were not found in patients whose conditions were diagnosed later in life. Men were significantly younger at diagnosis than were women. No obvious relation was found between HLA antigens and other clinical data. CONCLUSION: HLA-A26, B40, and DR9, which were found relatively frequently in the ancient Japanese population, seem to be associated with keratoconus in younger individuals. PMID- 11931793 TI - Clinicopathological report of cerebroocular myopathy syndrome. AB - PURPOSE: To report a case of cerebroocular myopathy syndrome with a focal absence of the Descemet membrane. DESIGN: Clinicopathological case report. METHODS: A clinical and histopathologic examination of eye, brain, and viscera of a white male newborn of normal term, who died at 11 days of age with the diagnosis of cerebroocular myopathy syndrome. RESULTS: The autopsy revealed hazy left cornea, hypoplasia of the cerebellum, and corpus collusum with mild microcephaly. Microscopy of the left eye showed the additional findings of a central focal defect of the Descemet membrane with an absence of the posterior third of central corneal stroma and delicate fibrous strands connected to the iris. CONCLUSION: The absence of the Descemet membrane and the missing posterior corneal keratocytes as well as the fibrous strands connecting the iris are findings to be added to the previously reported signs of cerebro-ocular myopathy. PMID- 11931794 TI - Rotational malposition during laser in situ keratomileusis. AB - PURPOSE: To investigate the degree of rotational malposition in eyes undergoing laser in situ keratomileusis. DESIGN: Prospective observational study. METHODS: We measured the rotational position of 240 eyes of 169 patients who underwent treatment for myopic or hyperopic astigmatism with the Alcon Summit Autonomous (Orlando, Florida) LADARVision excimer laser. Immediately preoperatively, each eye was marked while the patient was seated upright. Rotational position was measured on the supine patient immediately before beginning the laser exposure. RESULTS: For all 240 eyes, mean +/- standard deviation (SD) torsional misalignment was 4.1 +/- 3.7 degrees (right eye 3.8 +/- 3.7 degrees, left eye 4.2 +/- 3.6 degrees). A total of 20 eyes (8%) had a deviation of greater than 10 degrees. CONCLUSIONS: A 4 degree and 10 degree misalignment would theoretically result in a 14% and 35% undercorrection of astigmatism, respectively. Preoperative marking of the upright patient and subsequent rotational alignment of the supine patient before laser treatment may reduce the error in correction of astigmatism during excimer laser vision correction surgery. PMID- 11931795 TI - Resolution of epithelial ingrowth in a patient treated with 5-fluorouracil. AB - PURPOSE: To report the effect of 5-fluorouracil on the outcome of a single case of epithelial ingrowth secondary to a wound leak after a complicated penetrating keratoplasty. DESIGN: Case report. METHODS: A 44-year-old man with extensive epithelial ingrowth after trauma and a penetrating keratoplasty received a fluid gas exchange followed by two injections of 500 microg of 5-fluorouracil into the anterior chamber. RESULTS: Two weeks after receiving the 5-fluorouracil injections, complete disappearance of the epithelial ingrowth from the anterior chamber occurred. No evidence of recurrence was detectable at 5 months. CONCLUSION: Use of 5-fluorouracil is a potentially effective treatment of epithelial ingrowth, particularly in eyes with extensive involvement, in which the risks of traditional ablative therapeutic modalities are considerable. PMID- 11931796 TI - Accuracy and precision of intraocular injection volume. AB - PURPOSE: To assess the accuracy and precision of methods in clinical use for intraocular drug injection. DESIGN: Experimental laboratory investigation. METHODS: A total of 800 injections were performed using 20 different 1-ml syringes at intended volumes of 0.10 ml and 0.05 ml. The volume of each injection was calculated from mass of water delivered onto an analytic balance. RESULTS: The mean measured volume delivered was less than the intended volume for every syringe (P <.001 for each syringe at each volume). A total of 7% and 22% of injections differed by 20% or more from the intended volumes of 0.10 ml and 0.05 ml, respectively. Overdelivery of volume was uncommon. CONCLUSION: Clinically relevant materials and methods used for intraocular injection of drugs systematically underdelivered the intended volume. PMID- 11931797 TI - Acute posterior multifocal placoid pigment epitheliopathy and scleritis in a patient with pANCA-positive systemic vasculitis. AB - PURPOSE: To present acute posterior multifocal placoid pigment epitheliopathy and scleritis occurring in association with pANCA (perinuclear antineutrophil cytoplasmic antibody)-positive systemic vasculitis. DESIGN: Interventional case report. METHODS: A 69-year-old woman developed bilateral acute posterior multifocal placoid pigment epitheliopathy concurrent with rapidly progressive glomerulonephritis and bronchioalveolitis in both lungs. RESULTS: The patient showed a high titer of pANCA (myeloperoxidase-ANCA). She then developed scleritis in the left eye and multiple gastric ulcers. CONCLUSION: Acute posterior multifocal placoid pigment epitheliopathy and scleritis in this patient are manifestations that are part of pANCA-positive systemic vasculitis. This association further supports that acute posterior multifocal placoid pigment epitheliopathy is caused by vasculitic inflammation of the choriocapillaris. PMID- 11931798 TI - A new high-speed pneumatic vitrectomy cutter. AB - PURPOSE: To describe a new high-speed pneumatic vitrectomy instrument capable of 2000 cuts/minute. DESIGN: Observational report. METHODS: A high-speed guillotine vitrector, developed to perform vitrectomy, with the ability to adjust from 1 to 2000 cuts/minute, was used in 80 consecutive vitreoretinal cases with a variety of diagnoses in a hospital setting. RESULTS: The performance was almost vibration free. The instrument performed efficiently up to 1500 cuts/minute during vitrectomy, maintaining a constant flow rate. Higher cutting rates were used successfully for special indications. No complications were linked to the use of the instrument. CONCLUSION: The instrument was used successfully in 80 consecutive vitrectomies for a variety of diagnoses, performing efficiently up to 1500 cuts/minute, with a constant flow rate without complications. Higher cutting rates (2000 cuts/minute) were used successfully for special indications. PMID- 11931799 TI - Visual field defect caused by nerve fiber layer damage associated with an internal limiting lamina defect after uneventful epiretinal membrane surgery. AB - PURPOSE: To report a case of visual field defect caused by damage of the nerve fiber layer associated with an internal limiting lamina defect after uneventful epiretinal membrane peeling. DESIGN: Interventional case report. METHODS: In the right eye, a 43-year-old male patient developed a nasal step and mild inferior arcuate scotoma after uneventful epiretinal membrane surgery without any associated glaucoma. Transmission electron microscopy was performed on the epiretinal membrane specimens. RESULTS: An epiretinal membrane specimen showed adhesion between epiretinal membrane and axons of nerve fiber layer in the area of the internal limiting lamina defect. CONCLUSION: Adhesion between epiretinal membrane and retinal tissue in the area of the internal limiting lamina defect may cause damage of the nerve fiber layer and visual field defect after epiretinal membrane peeling. PMID- 11931800 TI - The effect of internal limiting membrane peeling on chronic recalcitrant pseudophakic cystoid macular edema: a report of two cases. AB - PURPOSE: To describe the effect of internal limiting membrane peeling in chronic recalcitrant pseudophakic cystoid macular edema. DESIGN: Two interventional case reports. METHODS: Two consecutive patients with chronic pseudophakic cystoid macular edema of 11-22 months duration, after uncomplicated cataract surgery, unresponsive to previous treatment, were enrolled. The surgical technique consisted of pars plana vitrectomy and internal limiting membrane peeling. Main outcome measures included best-corrected visual acuity, biomicroscopic appearance, and perifoveal leakage on fluorescein angiography. Optical coherence tomography scans were performed postoperatively. RESULTS: Pars plana vitrectomy with internal limiting membrane peeling resulted in anatomical, angiographic, and functional improvement in both patients, for total follow-up periods of 8 and 11 months, respectively. CONCLUSION: To our knowledge, this is the first report of chronic pseudophakic cystoid macular edema that favorably responded to pars plana vitrectomy with internal limiting membrane peeling, as concluded after a MEDLINE search. PMID- 11931801 TI - Photodynamic therapy increases the eligibility for feeder vessel treatment of choroidal neovascularization caused by age-related macular degeneration. AB - PURPOSE: To report angiographic observations about feeder vessel identification after photodynamic therapy in patients with choroidal neovascularization caused by age-related macular degeneration. DESIGN: Cohort study. METHODS: We analyzed fluorescein and indocyanine green dynamic angiography in 156 eyes of 145 patients before and after photodynamic therapy to identify the feeder vessels of the choroidal neovascular membrane. RESULTS: Before photodynamic therapy one or more feeder vessel could be detected in 35 (22.4%) out of 156 eyes with choroidal neovascularization. Three months after photodynamic therapy, a feeder vessel could be identified in 112 (84.2%) out of 133 eyes with persistent choroidal neovascularization. Among these, 16 eyes received direct laser photocoagulation of the feeder vessel and did not need any further photodynamic therapy. CONCLUSION: Previous photodynamic therapy improves the detection of the feeder vessel of the choroidal neovascularization. A sequential combined therapy (photodynamic and feeder vessel treatment) could be considered as an alternative to multiple photodynamic treatments. PMID- 11931802 TI - Enhanced S-cone syndrome with subfoveal neovascularization. AB - PURPOSE: To report a case of enhanced S-cone syndrome associated with subfoveal neovascularization. DESIGN: Observational case report. METHODS: A 23-year-old man, who was first examined at age 9 years, was found to have enhanced S-cone syndrome by clinical, electrophysiological, and molecular genetic examinations. RESULTS: At 9 years of age, a subfoveal neovascularization was present in his right eye and corrected visual acuity was RE: 0.15 and LE: 1.0. After he was 20 years old, cystoid changes in the macula of the left eye appeared and visual acuity, in the left eye, decreased from 1.0 to 0.02. CONCLUSION: The clinical course of enhanced S-cone syndrome is progressive, and we suggest that the subretinal neovascularization is a phenotypic variation of enhanced S-cone syndrome. PMID- 11931803 TI - Abnormal electroretinogram and abnormal electrooculogram after lightning-induced ocular injury. AB - PURPOSE: To report a case of abnormal electrooculogram and abnormal electroretinogram after lightning-induced ocular injury. DESIGN: Interventional case report. METHODS: A 39-year-old man was struck on the left forehead by a bolt of lightning. After resuscitation, he regained consciousness. RESULTS: Ophthalmic examination disclosed a best-corrected visual acuity of right eye (RE): 20/25 and left eye (LE): 20/50, burned eyelashes, punctate keratitis, iridocyclitis, anterior subcapsular lens opacity, missing foveolar reflex, and macular pigment epithelial defect LE. Fluorescein angiography revealed only an area of punctate leaking in the left eye. Electrooculogram showed reduced amplitudes, in the left eye. Arden ratio was RE: 2.04 and LE: 1.52. The amplitudes of scotopic and photopic electroretinogram b-waves were reduced in the left eye. After uneventful cataract surgery in the left eye, the follow-up electrooculogram and electroretinogram still showed reduced amplitude in the left eye. CONCLUSION: Lightning is an uncommon cause of ocular injury. This is the first report of abnormal electro-oculogram and abnormal electroretinogram after lightning-induced ocular injury. PMID- 11931804 TI - A rapidly growing choroidal melanoma. AB - PURPOSE: We describe a rapidly growing choroidal melanoma that failed to respond to consecutive globe-sparing treatments in an individual with ocular melanocytosis. DESIGN: Interventional case report. METHODS: A 53-year-old man with right ocular melanocytosis, decreased vision in the right eye, and right eye pain was referred for suspected choroidal melanoma. The lesion, which measured 2.4 mm x 6.6 mm x 6.0 mm, was treated with transpupillary thermotherapy on two occasions and with iodine-125 (I-125) plaque brachytherapy, both of which failed. Enucleation allowed histologic analysis. RESULTS: At enucleation, the choroidal melanoma in the right eye measured 13.6 mm x 16.5 mm x 16.9 mm, which demonstrated the most rapid growth and doubling time reported in a tumor of this type thus far. Pathologic analysis classified the tumor as a mixed-cell type. CONCLUSION: This case reiterates the association between ocular melanocytosis and choroidal melanoma, provides additional evidence that rapid growth and doubling time may be associated with transpupillary thermotherapy and I-125 plaque failure, and highlights the need to follow those tumors closely. PMID- 11931805 TI - Anterior ischemic optic neuropathy after a trans-Atlantic airplane journey. AB - PURPOSE: To report a case of anterior ischemic optic neuropathy after a trans Atlantic airplane journey. DESIGN: An observational case report. METHODS: A 48 year-old healthy man presented with severe visual loss in his left eye within 12 hours after a 15-hour-long trans-Atlantic airplane flight. The patient underwent slit-lamp examination, funduscopy, fluorescein angiography, automated perimetry, and various blood examinations. RESULTS: Visual acuity was LE: 20/30, and a mild left eye relative afferent pupillary defect was noted. Funduscopic evaluation revealed an edematous optic disk in the left eye with a para-diskal retinal hemorrhage. Fluorescein angiography revealed a leaking optic disk, and perimetry showed an inferior hemi-field scotoma. Anterior ischemic optic neuropathy was diagnosed. CONCLUSION: Prolonged airplane travel may compromise vision either by a thromboembolic mechanism caused by prolonged immobilization or by a vasospasm mechanism induced by the low oxygen pressure during the flight. PMID- 11931806 TI - Subperiosteal orbital hemorrhage as initial manifestation of Christmas disease (factor IX deficiency). AB - PURPOSE: To report a case of subperiosteal orbital and subgaleal hemorrhage with optic nerve compromise in a patient with a factor IX deficiency. DESIGN: Interventional case report. METHODS: A 5-year-old male presented 10 days after mild trauma with progressive left-sided scalp swelling, proptosis, and visual loss. RESULTS: The patient had marked proptosis of the left eye, 20/200 visual acuity, and an afferent pupillary defect. Magnetic resonance imaging demonstrated a large subgaleal and left subperiosteal orbital hematoma. Quantitative assays of coagulation proteins identified a factor IX deficiency (Christmas disease). CONCLUSION: Delayed-onset subgaleal and subperiosteal orbital hematoma can rarely be an initial manifestation of Christmas disease. PMID- 11931807 TI - Acrylic intraocular lens placement in conjunction with pars plana vitrectomy. PMID- 11931809 TI - Indocyanine green staining and removal of internal limiting membrane in macular hole surgery: histology and outcome. PMID- 11931810 TI - Mini-trabeculectomy as initial surgery for medically uncontrolled glaucoma. PMID- 11931813 TI - Autologous ipsilateral rotating penetrating keratoplasty. PMID- 11931816 TI - International ophthalmology strategic plan to preserve and restore vision-vision in the future. PMID- 11931834 TI - GTP-binding proteins in cell survival and demise: the emerging picture in the pancreatic beta-cell. AB - It is widely believed that guanine nucleotide-binding regulatory proteins (G proteins) play central roles as "molecular switches" in a variety of cellular processes ranging from signal transduction to protein and vesicle trafficking. To achieve these regulatory functions, G-proteins form complexes with a wide range of effector molecules whose activities are altered upon interaction with the G protein. These effector molecules can be either soluble or membrane bound, and it is likely that some are localized to secretory granules where they direct the movement, docking, and fusion of granules during exocytosis. The effector molecules regulated by G-proteins are diverse and include phospholipases, protein kinases, protein phosphatases, ion channels, adenylate cyclases, cytoskeletal elements, as well as secretory vesicle and plasma membrane-associated fusion proteins. The majority of studies performed in the pancreatic beta-cell have focused on the role of G-proteins in the regulation of insulin secretion, whereas very little attention has been focused on their potential involvement in other cellular processes. Such studies have identified and implicated both heterotrimeric (comprising alpha, beta, and gamma subunits) and monomeric (low molecular mass) G-proteins in the regulation of insulin secretion, but intriguing recent evidence has also begun to emerge which favors the view that they may be involved in the maintenance of beta-cell viability. In the present commentary, we will review this evidence and discuss the current understanding of the role of G proteins in the life and death of the beta-cell. PMID- 11931835 TI - Kappa-opioid receptor agonist inhibition of HIV-1 envelope glycoprotein-mediated membrane fusion and CXCR4 expression on CD4(+) lymphocytes. AB - Our previous studies have shown that the suppressive effect of kappa-opioid receptor (KOR) ligand treatment on HIV-1(AT) (a T-tropic strain) expression in acutely infected CD4(+) lymphocytes is time-dependent. This finding implied that the inhibition observed following treatment with KOR agonists such as U50,488 (trans-3,4-dichloro-N-methyl-N[2-(1-pyrolidinyl)cyclohexyl]benzeneaceamide methanesulfonate) occurs at an early step in the viral replication cycle, perhaps as early as viral entry. In the present study, we examined the hypothesis that U50,488 treatment of CD4(+) lymphocytes inhibits HIV-1 envelope (Env) glycoprotein-mediated membrane fusion. We used a vaccinia virus-based assay to measure the effects of U50,488 treatment of CD4(+) lymphocytes on HIV-1 IIIB Env glycoprotein-mediated fusogenic activity, based on the cytoplasmic activation of a reporter gene. The results show that U50,488 inhibited Env-mediated cell fusion in a bell-shaped concentration-response manner with suppression ranging between 31 and 98% at concentrations of 10(-8) and 10(-10)M (N=9 experiments). U50,488 was also found to inhibit cell fusion when monitored in situ with 5-bromo-4 chloro-3-indolyl-beta-D-galactopyranoside (X-gal) staining. Blockade of the inhibitory activity of U50,488 by the KOR antagonist nor-bialtorphimine (nor-BNI) suggested that U50,488 was acting via a KOR-related mechanism. Using flow cytometry, we demonstrated that the chemokine co-receptor CXCR4, but not CD4, is down-regulated as a consequence of KOR activation, with 44.2+/-3.5% suppression at 10(-10)M U50,488. These findings support the hypothesis that KOR-related activation of CD4(+) lymphocytes inhibits HIV-1 entry via down-regulation of CXCR4. PMID- 11931836 TI - Induction of arginase II in livers of bile duct-ligated rats. AB - Nitric oxide (NO) has been implicated in playing a role in liver cirrhosis, but the regulatory mechanisms are still unclear. As arginase shares a common substrate with NO synthase (NOS), the aim of this study was to investigate the expression of arginase I and II in cirrhotic liver. Liver cirrhosis was induced in rats by chronic bile duct ligation (BDL). Controls were sham-operated. Competitive polymerase chain reaction was performed to assay the expression of messenger RNA of arginase I and II. Protein expression was detected by immunohistochemistry and western-blotting. The level of arginine in plasma was lower in BDL rats, while the ornithine level in plasma was correspondingly higher (r= -0.96, P<0.0001). Arginase I messenger RNA was reduced significantly in BDL rats (3.34+/-0.32 vs. 1.32+/-0.21 x 10(4) attomole/microg of total RNA, sham vs. BDL, P<0.001), as well as arginase I protein. In contrast, arginase II mRNA was induced in the livers of BDL rats, with negligible expression in controls (0.35+/ 0.11 vs. 3.64+/-0.54 attomole/microg of total RNA, sham vs. BDL, P<0.001). Arginase II protein was localized in some hepatocytes and hyperplastic bile ductular epithelial cells of cirrhotic livers but not in control livers. In conclusion, arginase II was induced in BDL livers, while the expression of arginase I was down-regulated. These data suggest that arginase I and II are regulated differently and may have different functions in the livers of BDL rats. Reduction of arginase I in BDL livers may be responsible for the lowering of arginine levels in the plasma, while induction of arginase II could be important in regulating NO synthesis as well as other important mechanisms involved in liver cirrhosis. PMID- 11931837 TI - Cytoprotective role of taurine in a renal epithelial cell culture model. AB - Taurine (TAU) is a sulfur-containing amino acid that has been shown to decrease during aging and is believed to be important for cytoprotection. A decrease in TAU could exacerbate the accumulation of free radical-induced damage that may lead to cell death during the aging process. We have shown previously that TAU directly inhibits dopamine (DA) and (-)-3-(3,4-dihydroxyphenyl)-L-alanine (L dopa) oxidation. Experiments were conducted to establish a cytoprotective role for TAU. Porcine renal epithelial cells were treated for 1 hr with iron and catecholamines (L-dopa and DA) to produce cytotoxicity by a free radical and quinone mechanism in the absence and presence of 10 or 20mM TAU. Viability assays, protein, and DNA measurements were performed after a 24hr recovery period. In some experiments, cells were extracted immediately after the insult for DA and TAU content measurements using high performance liquid chromatography with electrochemical detection. Catecholamine-induced cytotoxicity caused a 50% loss in cell viability, and 10 or 20mM TAU provided significant protection from cytotoxicity and maintained the functional integrity of the cells. Photomicrographs showed attenuation in cell loss and swelling in the presence of TAU. Pretreatment with 1mM TAU followed by exposure to iron and L-dopa in the presence of 1mM TAU caused a moderate but non-significant increase in cell survival. These data conclusively show that TAU can play a cytoprotective role in the LLC-PK(1) cell culture model. PMID- 11931838 TI - A glutamate-gated chloride channel subunit from Haemonchus contortus: expression in a mammalian cell line, ligand binding, and modulation of anthelmintic binding by glutamate. AB - Glutamate-gated chloride channels (GluCls) are inhibitory ion channels that are sensitive to the antiparasitic drugs ivermectin (IVM) and moxidectin (MOX). We have transiently transfected COS-7 cells with a subunit of a GluCl (HcGluCla) from the parasitic nematode Haemonchus contortus. This subunit bound [3H]-IVM and [3H]-MOX with K(d) values of 0.11+/-0.021 and 0.18+/-0.02nM, respectively. Displacement analysis revealed that IVM and MOX bind to the same site on HcGluCla and that this site is likely distinct from the glutamate binding site. Glutamate was found to be an allosteric modulator of [3H]-MOX and [3H]-IVM binding and increased the affinity of [3H]-MOX for HcGluCla by more than 50% and that of [3H] IVM by more than 7-fold. These results point to both similarities and differences in the interactions of IVM and MOX with the GluCl. Aspartate, which is structurally similar to glutamate, had little or no effect on [3H]-IVM and [3H] MOX binding, suggesting that this ligand does not induce the conformational change necessary to potentiate macrocyclic lactone binding. These results also indicate that it may be possible to enhance the efficacy of macrocyclic lactone anthelmintics by administering these compounds with ligands acting allosterically to enhance their binding. PMID- 11931839 TI - Inhibition of type 1 protein phosphatase activity by activation of beta adrenoceptors in ventricular myocardium. AB - The regulation of protein phosphatase (PP) activity by cardiac beta-adrenergic receptor stimulation with isoproterenol (ISO) was studied in four groups of guinea pigs consisting of seven animals each. Group 1 received the vehicle solution only intraperitoneally; group 2, 6 microg/kg of ISO; group 3, 60 microg/kg of ISO; and group 4, 600 microg/kg of ISO. Total PP activity (consisting of both type 1 and type 2A PP), activity of each PP subtype, the cAMP dependent protein kinase activity ratio (-cAMP/+cAMP), the phosphorylation of PP inhibitor 1, and the phosphorylation of phospholamban were measured in ventricular tissue. PP activity was also studied in ventricular cardiomyocytes isolated from guinea pigs treated with and without 1 microM ISO or 1 microM ISO plus 10 microM propranolol, an antagonist of the beta-adrenoceptor. PP activity decreased significantly in membrane vesicles, but not in cytosolic fractions, of guinea pigs treated with 60 and 600 microg/kg of ISO compared with untreated animals. The PKA activity ratio, PLB phosphorylation, and PP inhibitor 1 phosphorylation increased in ventricles of guinea pigs treated with 60 and 600 microg/kg of ISO compared with vehicle-treated animals. The decrease in overall PP activity was due primarily to a reduction in type 1 but not type 2A PP activity. In isolated ventricular cardiomyocytes, PP activity was decreased significantly after treatment with 1 microM ISO, and this inhibition was reversed by treatment with 10 microM propranolol. The membrane vesicles of group 1 animals did not release any catalytic subunit of type 1 PP upon phosphorylation by exogenous PKA. These results indicate that activation of cardiac beta adrenoceptors inhibits type 1 PP activity via phosphorylation of PP inhibitor 1 in the ventricles. This effect is associated with the well-known effect of ISO on increases in the PKA activity ratio and PLB phosphorylation. Inhibition of type 1 PP activity could be one possible mechanism, in addition to activation of adenylate cyclase, by which ISO mediates enhanced contractility of the heart. PMID- 11931840 TI - Relative importance of maternal and embryonic microsomal epoxide hydrolase in 7,12-dimethylbenz[a]anthracene-induced developmental toxicity. AB - Microsomal epoxide hydrolase (mEH) catalyzes the hydrolysis of epoxide intermediates derived from drugs and environmental chemicals. The response of in vivo (embryo) and in vitro (embryo fibroblast) tests were analyzed using mEH-null and wild-type mice to determine the relative role of maternal and embryonic mEH in the developmental toxicity induced by 7,12-dimethylbenz[a]anthracene (DMBA). Embryos derived from DMBA-treated [50mg/kg, daily from gestational day (GD) 11 to GD 15] dams were analyzed. Although weight (P=0.0009) and crown-rump length (P=0.0003) of wild-type fetuses on GD 18 were significantly lower than those of mEH-null fetuses, respectively, no significant difference was found between mEH null and heterozygous fetuses of mEH-null dams. Cell viability was decreased to 50% in wild-type mouse embryo fibroblasts (MEFs) treated with 3 microM DMBA, but no significant decrease was found in mEH-null MEFs. DMBA-3,4-diol produced a significant decrease in cell viability and suppressed the proliferation of wild type MEFs at a 10-fold lower concentration than did DMBA. Although mEH protein was expressed in liver microsomes from wild-type embryos (GD 15), DMBA-3,4-diol was not detected among the DMBA metabolites. However, it was detected in the serum of wild-type pregnant mice treated with DMBA, but not in that of mEH-null mice. These results suggest that maternal mEH plays a major role in DMBA-induced developmental toxicity, and embryonic mEH is less involved in the toxicity. PMID- 11931841 TI - Bcl-2 family-mediated apoptotic effects of 3,3'-diindolylmethane (DIM) in human breast cancer cells. AB - 3,3'-Diindolylmethane (DIM) is a major in vivo derivative of the putative anticancer agent indole-3-carbinol (I3C), which is present in vegetables of the Brassica genus. At concentrations above 10 microM, DIM inhibited DNA synthesis and cell proliferation in both estrogen receptor replete (MCF-7) and deficient (MDA-MB-231) human breast cancer cells in a concentration- and time-dependent manner. These antiproliferative effects were accompanied by characteristic indications of programmed cell death in both cell lines, including externalization of phosphatidylserine, chromatin condensation, and DNA fragmentation. Furthermore, Western and Northern blot analyses, as well as coimmunoprecipitation assays, revealed that in both MCF-7 and MDA-MB-231 cells, DIM treatment decreased total transcript and protein levels of the apoptosis inhibitory protein Bcl-2, and the amount of Bcl-2 bound to the pro-apoptotic protein Bax. DIM treatment also caused an increase in Bax protein levels, but did not affect the level of Bax that was bound to Bcl-2. As a functional test of the role of Bcl-2 down-regulation in the DIM-induced apoptotic response, ectopic expression of Bcl-2 in MCF-7 cells was shown to attenuate the apoptotic effect of DIM. These results demonstrate that DIM can induce apoptosis in breast cancer cells independent of estrogen receptor status by a process that is mediated by the modulated expression of the Bax/Bcl-2 family of apoptotic regulatory factors. PMID- 11931842 TI - BGP-15 - a novel poly(ADP-ribose) polymerase inhibitor - protects against nephrotoxicity of cisplatin without compromising its antitumor activity. AB - Nephrotoxicity is one of the major dose limiting side effects of cisplatin chemotherapy. The antitumor and toxic effects are mediated in part by different mechanisms, thus, permitting a selective inhibition of certain side effects. The influence of O-(3-piperidino-2-hydroxy-1-propyl)nicotinic amidoxime (BGP-15) - a poly(ADP-ribose) polymerase (PARP) inhibitor - on the nephrotoxicity and antitumor efficacy of cisplatin has been evaluated in experimental models. BGP-15 either blocked or significantly reduced (60-90% in 100-200 mg/kg oral dose) cisplatin induced increase in serum urea and creatinine level in mice and rats and prevented the structural degeneration of the kidney, as well. The nephroprotective effect of BGP-15 treatment was revealed also in living mice by MRI analysis manifesting in the lack of oedema which otherwise developed as a result of cisplatin treatment. The protective effect was accompanied by inhibition of cisplatin-induced poly-ADP-ribosylation and by the restoration of the disturbed energy metabolism. The preservation of ATP level in the kidney was demonstrated in vivo by localized NMR spectroscopy. BGP-15 decreased cisplatin induced ROS production in rat kidney mitochondria and improved the antioxidant status of the kidney in mice with cisplatin-induced nephropathy. In rat kidney, cisplatin caused a decrease in the level of Bcl-x, a mitochondrial protective protein, and this was normalized by BGP-15 treatment. On the other hand, BGP-15 did not inhibit the antitumor efficacy of cisplatin in cell culture and in transplantable solid tumors of mice. Treatment with BGP-15 increased the mean survival time of cisplatin-treated P-388 leukemia bearing mice from 13 to 19 days. PARP inhibitors have been demonstrated to diminish the consequences of free radical-induced damage, and this is related to the chemoprotective effect of BGP 15, a novel PARP inhibitor. Based on these results, we propose that BGP-15 represents a novel, non-thiol chemoprotective agent. PMID- 11931843 TI - Characterization of the mechanism underlying stonustoxin-mediated relaxant response in the rat aorta in vitro. AB - Stonustoxin (SNTX) is a lethal factor isolated from the venom of the stonefish Synanceja horrida. Although SNTX exhibits a multitude of biological activities, the primary cause of death upon administration of the toxin is attributed to marked hypotension. We investigated the possible mechanisms underlying the vascular hyporeactivity of this novel toxin. Cumulative doses of SNTX (5-320 ng/mL) induced concentration-dependent relaxation in phenylephrine (PE)- precontracted rat aortic rings with intact endothelium. Endothelium removal abolished the relaxation induced by SNTX. Tetraethylammonium (TEA), an inhibitor of K(+) channels, partially inhibited SNTX-induced relaxation. Similarly, SNTX induced relaxation was partially attenuated by the SP receptor antagonist (NATB), whereas the inducible iNOS inhibitor, AMT-HCl, completely abolished the relaxation caused by SNTX. From the results obtained, it can be postulated that a component of SNTX-mediated vasorelaxation is via binding of either SNTX or SP to the SP receptors that are located on the endothelial cells. Occupation of these SP receptors causes subsequent production of NO and activation of K(+) channels, thus leading to vasorelaxation of the rat aortic rings. PMID- 11931844 TI - Apoptosis induced by extracellular ATP in the mouse neuroblastoma cell line N1E 115: studies on involvement of P2 receptors and adenosine. AB - Adenosine triphosphate (ATP) can be released in large amounts from (damaged) cells, leading to locally high concentrations. In this study, we investigated the effect of such high concentrations of ATP on neuroblastoma cells. ATP (>or=30 microM) induced apoptosis in the mouse neuroblastoma cell line N1E-115. Activation of the ATP receptor P2X(7) is one of the routes via which ATP has been shown to induce apoptosis. Although the P2X(7) receptor was present in N1E-115 cells, both at the protein and mRNA level, studies with the P2X(7) receptor agonist benzoyl-benzoyl ATP showed that this receptor was not involved in ATP induced apoptosis. It has been shown previously that adenosine induces apoptosis in N1E-115 cells after transport inside the cell. In this study, both dipyridamole, a nucleoside transport protein blocker, and uridine, a substrate for this transporter, were able to block ATP-induced apoptosis. This indicated that ATP had to be broken down to adenosine to induce apoptosis. The ecto nucleotidase inhibitors 6-N,N-diethyl-beta-dibromomethylene-D-adenosine-5' triphosphate (ARL67156) and alpha,beta-methylene adenosine 5'-diphosphate (AOPCP) commonly used to slow breakdown of ATP did not inhibit ATP breakdown appreciably, while the ATP antagonist PPADS inhibited the breakdown of AMP to adenosine; PPADS was also the only compound capable of inhibiting ATP-induced apoptosis. We conclude that the main route of ATP-induced apoptosis in N1E-115 cells was via breakdown to adenosine. PMID- 11931845 TI - Inhibition of rat lipoprotein oxidation after tetradecylthioacetic acid feeding. AB - We have previously shown that tetradecylthioacetic acid (TTA), a sulfur containing saturated fatty acid analogue, inhibits the oxidative modification of human low-density lipoprotein (LDL) in vitro. The oxidative modification of LDL is believed to be a crucial step in the progression of atherosclerosis. In the present study, we investigated the effect of TTA oral administration on the susceptibility of rat lipoprotein to undergo oxidative modification ex vivo. Lipoprotein resistance to copper-induced oxidation was highly improved after TTA administration to rats. Conjugated dienes produced after 150 min of lipoprotein oxidation were dramatically lowered in the TTA treated rats compared to controls. Malondialdehyde and lipid peroxides production by oxidation was highly limited. These effects were independent of any Vitamin E effects. More than 50% relative reduction in polyunsaturated fatty acids of the n-3 family, and more than 30% relative increase in 18:1n-9 fatty acid in the triacylglycerol (TAG)-rich lipoprotein were observed. TAG-rich lipoprotein lipids of TTA fed rats were decreased with more than 50% reduction in TAG. The data reported in this paper indicate a potent in vivo antioxidant capability of TTA that beside its hypolipidemic effect might be of importance in relation to the development of atherosclerosis. PMID- 11931846 TI - Lack of involvement of extracellular signal-regulated kinase (ERK) in the agonist induced endothelial nitric oxide synthesis. AB - In a recent paper, it was shown that stimulation of endothelial cells with bradykinin (BK) leads to phosphorylation of endothelial nitric oxide synthase (eNOS) mediated by extracellular signal-regulated kinase (ERK) (J. Biol. Chem. 275 (2000) 30707). Since in vitro phosphorylation by ERK reduced the catalytic activity of eNOS, it was suggested that this mechanism may be an important determinant of nitric oxide signalling in endothelial cells. To explore the physiological role of ERK as regulator of nitric oxide synthesis in intact cells, we measured the effects of the kinase inhibitor PD 98059 on BK- and ATP-induced nitric oxide formation in cultured endothelial cells and isolated vascular smooth muscle strips. PD 98059 completely inhibited ERK activation by BK and ATP in porcine aortic endothelial cells without affecting eNOS activation. Moreover, PD 98059 did not potentiate relaxation of isolated porcine pulmonary arteries to BK or ATP, indicating that ERK-catalysed eNOS phosphorylation does not contribute to the regulation of nitric oxide formation in intact cells or tissues. PMID- 11931847 TI - Characterization of anthracenediones and their photoaffinity analogs. AB - In an attempt to overcome the cardiotoxicity and cross-resistance problems caused by the anticancer drugs anthracyclines and anthracenediones during chemotherapy, we have developed a series of aza-anthracenedione compounds by modifying the chromophore and the side arms of anthracyclines and anthracenediones. One of these aza-anthracenediones, 6,9-bis[(2-aminoethyl)amino]benzo[g]isoquinoline-5,10 dione (BBR 2778), which is currently under phase II clinical trials, showed remarkable antitumor activity and appeared to lack a cardiotoxic effect in preclinical studies. However, it was still cross-resistant against multidrug resistance (MDR) cells expressing P-glycoprotein (P-gp). In contrast, another aza anthracenedione, 6,9-bis[[2-(dimethylamino)ethyl]amino]benzo[g]isoquinoline-5,10 dione, which has side arm structures different from those of BBR 2778, was highly active against MDR cells. In this study, BBR 2778, BBR 2378, and an anthracenedione compound, 1,4-bis[(2-aminoethyl)amino]-5,8-dimethyl-9,10 anthracenedione, were used to assess the relationship between the chemical structures of these drugs and their interactions with DNA and P-gp. In addition, the biological and pharmacological influences of photoaffinity labeling were also studied for BBR 2778 and DEH. As the results indicate, the photolabeled analogs of BBR 2778 and DEH were less DNA-reactive and less cytotoxic. The more lipophilic compound, BBR 2378, and the photolabeled analogs of BBR 2778 and DEH inhibited P-gp labeling by azidopine better than did the more hydrophilic parental compounds. These studies suggested that the DNA binding affinity of BBR 2778 and DEH could be important in determining their cytotoxicity, and that the chemical structure of the side arms and the lipophilicity of these drugs are critical in determining their cross-resistance. PMID- 11931848 TI - Overexpression of sorcin, a calcium-binding protein, induces a low level of paclitaxel resistance in human ovarian and breast cancer cells. AB - Paclitaxel, an antimitotic, anticancer agent, induces cell cycle arrest in the mitotic phase by binding to the beta-tubulin subunit and forming highly stable microtubule polymers that resist depolymerization. The overexpression of the P glycoprotein (P-gp) and/or alteration in the cellular microtubules is associated with the development of paclitaxel resistance. However, we have established a paclitaxel-resistant human ovarian carcinoma subline (2008/13/4) wherein the degree of resistance could not be correlated with overexpression of P-gp, alterations in the alpha- and beta-tubulin isotypes, or changes in the drug binding affinity of the microtubules. mRNA differential display analysis revealed the overexpression of sorcin, a calcium-binding protein in the 2008/13/4 cells. However, no detectable changes in the intracellular calcium levels were detected in the parental and the paclitaxel-resistant variant. Furthermore, co-treatment with A23187, a calcium ionophore, did not alter the cytotoxicity of paclitaxel against the parental and the paclitaxel-resistant cells. Transfection of the parental 2008 cells with full-length sorcin cDNA induced a low level (3-5-fold) of paclitaxel resistance. In addition, transfection of human breast cancer cells with the full-length sorcin cDNA also led to the induction of a low level of paclitaxel resistance in the transfectants. Although the overexpression of sorcin did not produce high levels of paclitaxel resistance, the results obtained present compelling evidence of the involvement of sorcin in developing low-level paclitaxel resistance in a variety of tumor cells. The precise biochemical mechanism(s) by which sorcin overexpression induces low-level paclitaxel resistance is currently under investigation. PMID- 11931849 TI - The distribution of copper, zinc- and manganese-superoxide dismutase, and glutathione peroxidase messenger ribonucleic acid in rat basal ganglia. AB - Oxidative stress may contribute to the progression of Parkinson's disease, and while the status of antioxidant enzymes is thus important, little data on their regional distribution in basal ganglia exist. We now report on the distribution and levels of messenger ribonucleic acid (m-RNA) for the antioxidant enzymes copper, zinc-superoxide dismutase (Cu,Zn-SOD), manganese-superoxide dismutase (Mn SOD), and glutathione peroxidase in rat basal ganglia using in situ hybridisation histochemistry with complementary deoxyribonucleic acid probes specific for these enzymes. The m-RNA for Cu,Zn-SOD, Mn-SOD, and glutathione peroxidase was expressed throughout basal ganglia. Levels of m-RNA were significantly higher in substantia nigra pars compacta than in all other regions of basal ganglia for both Cu,Zn-SOD (53-62%, P<0.001) and Mn-SOD (37-45%, P<0.05). Mn-SOD m-RNA levels were also significantly higher in SN pars reticulata than in the nucleus accumbens (10%, P<0.05) and striatum (12%, P<0.01). In contrast, glutathione peroxidase m-RNA levels were only significantly higher in SN pars compacta when compared with SN pars reticulata (23%, P<0.05), and in the striatum when compared with the nucleus accumbens (21%, P<0.05). The data suggest that SN pars compacta may be vulnerable to oxidative stress and thus dependent on the high antioxidant capacity provided by these cytoprotective enzymes. In conclusion, this study demonstrates the relative distribution of antioxidant enzymes in rat basal ganglia and forms the basis for further study in rodent models of Parkinson's disease. PMID- 11931850 TI - Structure-activity relationships for inhibition of human 5alpha-reductases by polyphenols. AB - The enzyme steroid 5 alpha-reductase (EC 1.3.99.5) catalyzes the NADPH-dependent reduction of the double bond of a variety of 3-oxo-Delta(4) steroids including the conversion of testosterone to 5 alpha-dihydrotestosterone. In humans, 5 alpha reductase activity is critical for certain aspects of male sexual differentiation, and may be involved in the development of benign prostatic hyperplasia, alopecia, hirsutism, and prostate cancer. Certain natural products contain components that are inhibitors of 5 alpha-reductase, such as the green tea catechin (-)-epigallocatechin gallate (EGCG). EGCG shows potent inhibition in cell-free but not in whole-cell assays of 5 alpha-reductase. Replacement of the gallate ester in EGCG with long-chain fatty acids produced potent 5 alpha reductase inhibitors that were active in both cell-free and whole-cell assay systems. Other flavonoids that were potent inhibitors of the type 1 5alpha reductase include myricetin, quercitin, baicalein, and fisetin. Biochanin A, daidzein, genistein, and kaempferol were much better inhibitors of the type 2 than the type 1 isozyme. Several other natural and synthetic polyphenolic compounds were more effective inhibitors of the type 1 than the type 2 isozyme, including alizarin, anthrarobin, gossypol, nordihydroguaiaretic acid, caffeic acid phenethyl ester, and octyl and dodecyl gallates. The presence of a catechol group was characteristic of almost all inhibitors that showed selectivity for the type 1 isozyme of 5 alpha-reductase. Since some of these compounds are consumed as part of the normal diet or in supplements, they have the potential to inhibit 5 alpha-reductase activity, which may be useful for the prevention or treatment of androgen-dependent disorders. However, these compounds also may adversely affect male sexual differentiation. PMID- 11931851 TI - The effects of natural ligands of hormone receptors and their antagonists on telomerase activity in the androgen sensitive prostatic cancer cell line LNCaP. AB - The effects of the 17beta-estradiol, dihydrotestosterone and hormone antagonists tamoxifen and bicalutamide on telomerase activity and expression of cell cycle related proteins in the androgen-sensitive prostatic cancer cell line LNCaP were studied. The cell line was grown in RPMI supplemented with 2.5% charcoal-stripped FBS for 72 hr. The IC(50) of tamoxifen and bicalutamide and the optimal stimulatory concentrations of 17beta-estradiol and dihydrotestosterone were determined by means of the cell-viability assay, the activity of telomerase was measured by the telomere repeat amplification protocol (TRAP) and the expression of proteins was analysed by the Western blot technique. 17beta-estradiol stimulated cell growth more effectively than dihydrotestosterone whereas hormone antagonists tamoxifen and bicalutamide caused a significant decrease in cell viability. The treatment of cells by a combination of low doses of 17 beta estradiol and dihydrotestosterone stimulated cells stronger than treatment by a single hormone. Only 17beta-estradiol, in concentration of 10nM, increased strongly the expression of p21(Waf1/Cip1) and increased slightly telomerase activity in the LNCaP cells. 50 microM of bicalutamide down-regulated the levels of the androgen receptor, the proliferating cell nuclear antigen and telomerase activity, and up-regulated the expression of p27(Kip1). We hereby describe the first observation of the influence of bicalutamide on telomerase activity and a positive correlation between the effect of 17beta-estradiol and the induction of both the endogenous cyclin-dependent kinase inhibitor, p21(Waf1/Cip1), and telomerase activity in a prostatic cancer cell line LNCaP. These findings can shed a new light on the steroid-signaling pathway in prostate cancer cells. PMID- 11931852 TI - Inhibition of inducible prostaglandin E(2) synthase by 15-deoxy-Delta(12,14) prostaglandin J(2) and polyunsaturated fatty acids. AB - Prostaglandin E(2) synthase (PGE synthase) is one of the membrane-associated proteins in the eicosanoid and glutathione metabolism (MAPEG) family of microsomal enzymes and constitutes a novel inducible enzyme involved in inflammation and pyretic responses. We report, using a reversed-phase HPLC assay for the production of tritiated prostaglandin E(2) (PGE(2)) by membranes from cells overexpressing human microsomal PGE synthase, that PGE synthase activity is inhibited effectively by 15-deoxy-Delta(12,14)-prostaglandin J(2) and arachidonic acid. The anti-inflammatory compound 15-deoxy-PGJ(2) was considerably more potent at inhibiting PGE synthase (IC(50)=0.3 microM) than the closely related PGJ(2) or Delta(12)-PGJ(2), or the reaction product PGE(2). Arachidonic acid, docosahexaenoic acid, and eicosapentaenoic acid inhibited PGE synthase with a similar potency (IC(50)=0.3 microM) and were more potent inhibitors than various fatty acid analogues. The present results on the inducible PGE synthase extend observations on the ability to bind arachidonic acid to another member of the MAPEG family, and also suggest a novel mechanism of action for the anti inflammatory effects of DHA, EPA, and 15-deoxy-PGJ(2). PMID- 11931853 TI - Hypoxia/reoxygenation-induced damage to mitochondrial activity is determined by glutathione threshold in astroglia-rich cell cultures. AB - It has been shown that astrocytes play an important role during ischemia/reperfusion and in neurodegenerative diseases by supporting neuronal functions, but the effect of these pathophysiological conditions on this particular cell type is still unclear. Here, we investigated the ischemia/reperfusion-induced damage to astroglia-rich cells. For that purpose, we studied the effects of substrate deprivation and hypoxia/reoxygenation on total cellular glutathione contents, and mitochondrial function. Substrate deprivation as well as increasing time of cultivation in vitro (from 2 to 4 weeks) induced a decrease in the total glutathione content. Three qualitative distinct concentration ranges of the glutathione pool with respect to the effect of hypoxia/reoxygenation on the glutathione content were found: (i) high glutathione levels above 40 nmol per mg protein remained unchanged during hypoxia/reoxygenation. (ii) Hypoxia/reoxygenation was accompanied by higher glutathione levels in comparison to controls at intermediate initial glutathione concentrations of about 20 up to 40 nmol per mg protein. (iii) Below an initial glutathione threshold concentration of about 20 nmol per mg protein, hypoxia/reoxygenation led to a stronger decrease of glutathione levels in comparison to controls. Decrease of mitochondrial respiratory chain activity during hypoxia/reoxygenation only occurred at low initial glutathione concentrations below 20 nmol per mg protein. Our data emphasize the important role of glutathione with respect to the defense of mitochondria against oxidative stress in astroglia cells during hypoxia/reoxygenation. PMID- 11931855 TI - Fos expression induced by warming the preoptic area in rats. AB - The preoptic area (POA) occupies a crucial position among the structures participating in thermoregulation, but we know little about its efferent projections for controlling various effector responses. In this study, we used an immunohistochemical analysis of Fos expression during local warming of the preoptic area. To avoid the effects of anesthesia or stress, which are known to elicit Fos induction in various brain regions, we used a novel thermode specifically designed for chronic warming of discrete brain structures in freely moving rats. At an ambient temperature of 22 degrees C, local POA warming increased Fos immunoreactivity in the supraoptic nucleus (SON) and the periaqueductal gray matter (PAG). Exposure of animals to an ambient temperature of 5 degrees C induced Fos immunoreactivity in the magnocellular paraventricular nucleus (mPVN) and the dorsomedial region of the hypothalamus (DMH). Concurrent warming of the POA suppressed Fos expression in these areas. These findings suggest that thermal information from the preoptic area sends excitatory signals to the SON and the PAG, and inhibitory signals to the mPVN and the DMH. PMID- 11931854 TI - Involvement of brain cytokines in the neurobehavioral disturbances induced by HIV 1 glycoprotein120. AB - Intracerebroventricular (i.c.v.) administration of HIV-1 glycoprotein 120 (gp120), the envelope protein used by the virus to gain access into immune cells, induces neurobehavioral alterations in rats. To examine the role of proinflammatory cytokines in mediating these effects, we measured the effects of gp120 on brain proinflammatory cytokine expression and the effects of anti inflammatory agents, including interleukin-1 receptor antagonist (IL-1ra), pentoxifylline (a TNFalpha synthesis blocker) and IL-10, on gp120-induced sickness behavior. I.c.v. administration of gp120 induced the expression of IL 1beta, but not TNFalpha, mRNA in the hypothalamus, 3 h after the injection. Pretreatment of rats with IL-1ra, but not with pentoxifylline, significantly attenuated gp120-induced anorexia and loss in body weight, whereas both agents had no effect on gp120-induced reduction in locomotor activity in the open field. Pretreatment with either IL-1ra and pentoxifylline simultaneously, or with IL-10, produced effects that were similar to the effects of IL-1ra alone. Together, these findings indicate that IL-1, but not TNFalpha, mediates some of the behavioral effects of acute gp120 administration, suggesting that brain IL-1 may be involved in some of the neurobehavioral abnormalities evident in AIDS patients. PMID- 11931856 TI - In situ localization of nitric oxide synthase and direct evidence of NO production in rat retinal ganglion cells. AB - The expression of isoforms of nitric oxide synthase (NOS), enzymes responsible for NO production, and the synthesis of nitric oxide (NO) in rat retinal ganglion cells (RGCs) during synaptogenesis for various phases of the pre- and postnatal developmental periods were investigated. The retinas from prenatal, lactating, young, and adult rats were fixed in paraformaldehyde. The cryosections or paraformaldehyde-fixed ganglion cells purified from rat pups were immunostained for constitutive isoforms of NOS (n and eNOS) and observed with a confocal laser scanning microscope. Synthesis of NO in the RGCs was achieved by in vitro stimulation with glutamate. The intracellular NO levels were measured in real time using diaminofluorescein-2 diacetate, a fluorescence indicator of NO. Immunohistochemical analysis revealed nNOS and eNOS expressed in retinal ganglion cells during the first 2 postnatal weeks. Cultured RGCs also expressed nNOS and eNOS in vitro. Intracellular NO levels in cultured RGCs showed spontaneous fluctuation during a 20-min observation. The presence of both a non-specific NOS inhibitor, L-NAME, and a specific nNOS inhibitor, 7-NI, significantly inhibited (P<0.001) the increase of intracellular NO 6 and 8 min after the introduction of L-arginine and glutamate to the medium. This study revealed that all constitutive NOS isoforms are expressed in RGCs and demonstrated that NO is produced by nNOS mainly through stimulation by glutamate in cultured RGCs. PMID- 11931857 TI - Chronic treatment with supraphysiological levels of corticosterone enhances D MDMA-induced dopaminergic neurotoxicity in the C57BL/6J female mouse. AB - Chronic stress and extended periods of elevated circulating glucocorticoids have been reported to exacerbate excitotoxicity-induced hippocampal neuronal injury in rat. Despite continued interest in the effects of protracted exposure to stress or glucocorticoids, there has been little examination of how other types of neurotoxicity may be exacerbated or blocked, by stress. Here we examined the effects of chronic supraphysiologic levels of corticosterone on D-3,4 methylenedioxymethamphetamine (D-MDMA)-induced striatal dopaminergic neurotoxicity in the female C57BL/6J mouse. Corticosterone (5 mg, 15 mg or placebo) pellets were implanted to continuously elevate circulating glucocorticoids and create a model of the ultimate effect of chronic activation of the hypothalamic-pituitary-adrenal axis. After 7 days, a neurotoxic regimen of D-MDMA was administered (20 mg/kg s.c. every 2 hx4); thymus, spleen, striatum and hippocampus were collected 72 h later. Significant involution of thymus and spleen confirmed the bioavailability of the corticosterone at both dosages. D MDMA increased the striatal levels of the astrocyte-localized protein glial fibrillary acidic protein (GFAP, a marker of gliosis); both dosages of corticosterone exacerbated this increase but only the 15 mg pellet exacerbated the decrease in tyrosine hydroxylase protein. Corticosterone alone or in combination with D-MDMA produced no neural injury in hippocampus, as measured by GFAP. Our work indicates corticosterone was able to increase the vulnerability of the striatum, but not the hippocampus to D-MDMA. An examination of other mouse strains and models of neurotoxic injury would be useful in determining the general validity of the glucocorticoid neuroendangerment hypothesis. PMID- 11931858 TI - Chronic social stress during puberty enhances tyrosine hydroxylase immunoreactivity within the limbic system in golden hamsters. AB - The present study was carried out to determine the effects of chronic exposure to social stress during puberty on the dopamine system in male golden hamsters. Experimental animals were socially subjugated between postnatal days 28 (P28) and 42. All animals were sacrificed on P46 and their brains processed for immunocytochemistry to tyrosine hydroxylase (TH). A large increase in the number of TH-immunoreactive (TH-ir) neurons was noted within the posterior portion of the medial amygdaloid nucleus and the posterior portion of the medial division of the bed nucleus of the stria terminalis in subjugated animals as compared to controls. This effect appeared to be site-specific as no difference was seen between groups in the periventricular nucleus, another steroid receptor-rich area. The data suggest that these dopamine neurons may play an important role in the behavioral changes associated with chronic social stress during puberty. PMID- 11931859 TI - Heroin sensitization as mapped by c-Fos immunoreactivity in the rat striatum. AB - Immunohistochemistry was used to map the induction of c-Fos protein in the forebrain of rats treated with heroin. Acute injection of heroin to drug-naive rats caused significant induction of c-Fos protein in the nucleus accumbens shell, whereas the same dose of heroin given to drug-sensitized rats significantly increased c-Fos immunoreactivity in the dorsomedial caudate putamen. These results show that the heroin-induced pattern of c-Fos protein in the rat striatum differs according to the rat's drug history. These findings may represent a neural correlate of the motor components of heroin sensitization. PMID- 11931861 TI - Prenatal protein malnutrition decreases mossy fibers-CA3 thorny excrescences asymmetrical synapses in adult rats. AB - Prenatal protein malnutrition has deleterious effects on hippocampal structure and function that likely result from decreased synapse number. We thus evaluated long-term effects of prenatal protein malnutrition on the mossy fibers-CA3 thorny excrescences asymmetrical synapses in 220-day-old rats. Protein malnourished rats born from pregnant dams fed with 6% casein diet were cross-fostered to lactating control rats at birth. Control animals were fed with a 25% casein diet. Timm's stained material was used to estimate the total reference volume of the mossy fiber system suprapyramidal bundle by means of stereology. The mossy fiber-CA3 asymmetrical synapse numerical density was obtained by electron microscopy, using the physical disector method. The total number of mossy fiber-CA3 asymmetrical synapses was determined on the basis of the total reference volume of the mossy fiber system suprapyramidal bundle and the mossy fiber-CA3 asymmetrical synapse numerical density. Prenatal protein malnutrition produced long-lasting, significant decreases in the volume of the mossy fiber system suprapyramidal bundle and in the numerical density of mossy fiber-CA3 asymmetrical synapse, suggesting a reduction in the total number of this synapse type. Hence, prenatal protein malnutrition induces long lasting deleterious effects on the progression of developmental programs controlling synaptogenesis and/or synaptic consolidation, likely by affecting a myriad of cellular processes. PMID- 11931860 TI - d-MDMA during vitamin E deficiency: effects on dopaminergic neurotoxicity and hepatotoxicity. AB - The mechanism of 3,4-methylenedioxymethamphetamine (d-MDMA)-induced neurotoxicity may involve formation of toxic radical species. Endogenous defenses against toxic radical species include tissue stores of vitamin E, and thiols. We examined whether vitamin E deficiency could alter d-MDMA-induced neurotoxicity by administration of the drug to animals with diet induced vitamin E deficiency. Brain vitamin E levels in deficient mice were reduced 75% compared to sufficient animals. Animals received d-MDMA 5 or 10 mg/kg or saline (delivered every 2 hx4, s.c.). Diet slightly altered d-MDMA-induced temperature modulation. In brain, MDMA treatment reduced vitamin E, total antioxidant reserve and protein thiols 72 h after the first dose. In liver, MDMA treatment reduced glutathione and total antioxidant reserve at the same time point. The vitamin E-deficient group, treated with the low dose of d-MDMA, exhibited neurotoxic responses, including reduced striatal dopamine (47%) and elevated GFAP protein (3-fold): while the sufficient diet group was not altered. The higher d-MDMA dose caused neurotoxic responses in both diet groups. Liver toxicity was determined by histopathologic examination. d-MDMA caused hepatic necrosis that was more severe in vitamin E deficient than sufficient mice. These data indicate that (1) d-MDMA administration reduces antioxidant measures at a time coincident with d-MDMA induced neuronal damage and (2) vitamin E deficiency increases susceptibility to d-MDMA-induced neurotoxicity and hepatic necrosis. PMID- 11931862 TI - Combining neural network predictions for medical diagnosis. AB - We present our results from combining the predictions of an ensemble of neural networks for the diagnosis of hepatobiliary disorders. To improve the accuracy of the diagnosis, we train the second level networks using the outputs of the first level networks as input data. The second level networks achieve an accuracy that is higher than that of the individual networks in the first level. Compared to the simple method which averages the outputs of the first level networks, the second level networks are also more accurate. We discuss how the overall predictive accuracy can be improved by introducing bias during the training of the level one networks. PMID- 11931863 TI - The effects of automatic artifact rejection on evoked potential recordings. AB - An analysis of the effects of automatic artifact rejection on the evoked potential recordings is made using both a theoretical analysis and computer simulations. It is demonstrated that when the noise at each measurement point is independently distributed with maximum probability near zero amplitude, the bias and changes in variance induced by the automatic artifact rejection are small unless more than 80% of traces are rejected. When high-amplitude noise transients occur, there can be significant bias in the evoked potential when the artifact rejection threshold is set near the amplitude of the noise transients. Even more substantial distortions in the evoked potential occur when the noise is not independently distributed at each measurement point. It is important to recognize the possibility of these effects during clinical evoked potential recording. PMID- 11931864 TI - Tracking rhythmicity in nonstationary quasi-periodic biomedical signals using adaptive time-varying covariance. AB - A time-varying covariance method for detecting and quantifying the evolution of rhythmicity (frequency) in persistently varying quasi-periodic nonstationary signals is presented. The basic method, evaluated using chirp signals, utilizes a shifting window of fixed length. A substantial reduction in estimation bias and variability are obtained by utilizing an adaptive window whose length is dependent on past frequency estimates. The adaptive window yields estimates that are comparable in accuracy to those obtained using high-resolution time-frequency representation but with lower computation requirements and the potential for on line application. Finally, an example of the application of the method for analyzing a neural recording is also illustrated. PMID- 11931865 TI - Remodeled-matrix contraction by fibroblasts: numerical investigations. AB - It is well known that the fibroblast-collagen-matrix contraction model is a unique way to study mechanical interactions that regulate wound contraction of connective tissue cells. This contraction, due to cell traction, plays important roles in wound healing and pathological contractures. A continuum model initially used for the study of mesenchymal morphogenesis is revisited and numerically investigated by assuming that the extracellular matrix has adaptive-elastic properties. The set of non-linear partial differential equations is solved numerically by a finite difference method and the obtained results are discussed. PMID- 11931866 TI - Computing DIT from energy expenditure measures in a respiratory chamber: a direct modeling method. AB - The possibility of computing Diet Induced Thermogenesis (DIT) is an important feature of metabolic investigations. However, methodological problems have affected the determination of DIT in the indirect calorimetric chamber. DIT has been commonly estimated by regressing energy expenditure on a measure of physical activity. Although used for many years as the only feasible approach to calculate DIT in a respiratory chamber, this traditional method has been criticized because of an apparent underestimation of the DIT, but no alternative method has been suggested so far. The present work proposes to estimate DIT directly by means of a mathematical model. This approach also allows to simultaneously estimate other parameters, namely resting energy expenditure (REE), physical activity (PA) and physical exercise (PE). PMID- 11931867 TI - Abstracts of the 50th Annual Meeting of the Pacific Coast Reproductive Society. April 17-21, 2002. Rancho Mirage, California, USA. PMID- 11931915 TI - Horizontal space misrepresentation in unilateral brain damage. I. Visual and proprioceptive-motor influences in left unilateral neglect. AB - Patients with unilateral neglect can misperceive horizontal distances in the contralesional space as being shorter than equivalent ipsilesional ones. We evaluated the visual and motor-proprioceptive components of space misrepresentation through a distance reproduction task performed both with and without visual guidance. Four groups of right brain damaged patients (neglect with hemianopia (N+H+), neglect with inferior quandrantanopia (N+Q+), neglect without hemianopia (N+H-) and patients without neglect or hemianopia (N-H-)) and one group of healthy controls (C) performed the line bisection task and reproduced horizontal distances either by setting the endpoints or by doubling the length of a line in the contralesional or ipsilesional space. The doubling length task was administered in three different conditions: (a) visuomotor (the patient draws the line in free vision); (b) visual (by sight the patient guides the examiner drawing the line); (c) proprioceptive-motor (the patient is blindfolded and manually inspects and extends the horizontal distance subtended by the line). Compared to C and N-H- patients, only N+H+ patients exhibited a significant ipsilesional shift in line bisection. N+H+ patients showed the most severe contralesional-overextension/ipsilesional-underextension asymmetry in the endpoint, visuomotor and visual line extension task. In the proprioceptive-motor condition no asymmetry was found and N+H- showed greater overextension on both sides of space. In N+H-, brain damage was mainly centered in central-frontal cortex and basal ganglia. These findings re-emphasize the relevance of damage to visual retinotopically organized representations of space in the genesis of horizontal space misrepresentation of neglect patients and suggest the possible association of a non-lateralized defective processing of proprioceptive-motor information with unilateral neglect. PMID- 11931916 TI - Horizontal space misrepresentation in unilateral brain damage. II. Eye-head centered modulation of visual misrepresentation in hemianopia without neglect. AB - We used a visual distance reproduction task (endpoint task) to evaluate horizontal space representation in two left brain damaged (LBD) and three right brain damaged (RBD) patients with contralateral homonymous hemianopia and no neglect. All patients were examined in the chronic phase of the stroke and were aware of their visual field defect. Along with contralesional deviation in the line bisection task, all patients estimated size (Landmark task) and distances in the contralesional space as being longer than equivalent size and distances located in the ipsilesional space. Misreproduction of distances was abolished or reduced when the task was performed in the ipsilesional head-centred space with the head turned contralesionally. This finding points out that misrepresentation of horizontal space linked to hemianopia can be modulated by combined proprioceptive input from eye and neck muscles. The pattern of misrepresentation found in chronic hemianopic patients is opposite to the one described in chronic neglect patients with concomitant hemianopia. These different patterns of space misrepresentation are the likely consequence of the presence, in hemianopics, and the absence, in neglect patients with hemianopia, of compensatory strategies based on the non-retinotopic and multimodal coding of spatial positions falling in the retinotopically organised blind field. PMID- 11931917 TI - Reading the mind from eye gaze. AB - Baron-Cohen [Mindblindness: an essay on autism and theory of mind. Cambridge, MA: MIT Press, 1997] has suggested that the interpretation of gaze plays an important role in a normal functioning theory of mind (ToM) system. Consistent with this suggestion, functional imaging research has shown that both ToM tasks and eye gaze processing engage a similar region of the posterior superior temporal sulcus (STS). However, a second brain region associated with ToM, the medial prefrontal (MPF) cortex, has not been identified by previous eye gaze studies. We discuss the methodological issues that may account for the absence of MPF activation in these experiments and present a PET study that controls for these factors. Our experiment included three conditions in which the proportions of faces gazing at, and away from, the participant, were as follows: 100% direct [0% averted], 50% direct-50% averted, and 100% horizontally averted [0% direct]. Two control conditions were also included in which the faces' gaze were averted down, or their eyes were closed. Contrasts comparing the gaze conditions with each of the control conditions revealed medial frontal involvement. Parametric analyses showed a significant linear relationship between increasing proportions of horizontally averted gaze and increased rCBF in the MPF cortex. The opposite parametric analysis (increasing proportions of direct gaze) was associated with increased rCBF in a number of areas including the superior and medial temporal gyri. Additional subtraction contrasts largely confirmed these patterns. Our results demonstrate a considerable degree of overlap between the medial frontal areas involved in eye gaze processing and theory of mind tasks. PMID- 11931918 TI - Object motor representation and reaching-grasping control. AB - The following two competing hypotheses were tested in the present study. Is grasp guided by multiple representations of a single object, each of which codes a different grasp motor act according to the physical properties of that item? Conversely, is grasp guided by a single representation that codes all the possible affordances enabled by the object? Subjects reached different objects, but the object part used by subjects to grasp them was identical. In experiments 1 and 2, two familiar objects (fruits) which varied for size and shape were presented. Subjects grasped their stalks whose size and shape were equal. In experiments 3-7 the presented objects were geometrical solids, which varied, respectively, for weight, volume, intrinsic height, centre of mass and shape. Nevertheless, in all experiments the object portion where subjects' fingers grasped it had the same physical features. Finally, experiment 8 was a control experiment in which subjects reached and grasped equal handles of bells of the same shape, but different size. Volume, shape, and familiarity of the object influenced the grasp kinematics, even if the features of the grasped object part did not change. Variation in intrinsic object height and weight influenced final reach kinematics. Variation in centre of mass influenced neither grasp nor reach kinematics. Data are discussed in support of the hypothesis that a single object motor representation, which codes all the object affordances, is involved in grasp kinematic implementation. PMID- 11931919 TI - Phonology and neuropsychology of the English past tense. AB - The double dissociation between the regular and irregular past tense in English has been explained in terms of dual and single mechanism accounts. In previous research we have argued that problems with the regular past tense in patients with left inferior frontal damage arise from morpho-phonological parsing difficulties [Trends in Cognitive Science 2 (1998) 428]. This claim has recently been challenged by a single mechanism connectionist account which argues that a general phonological processing deficit causes the poor performance on the regular past tense, with morphological factors playing no explicit role [Proceedings of the National Academy of Sciences 96 (1999) 7592]. We used a speeded judgement task with four patients who have documented difficulties with the regular past tense to contrast the claims made by these different approaches. We compared patients' ability to detect the difference between the past tense and stem of regular (hugged/hug) and irregular (taught/teach) past tense verbs, as well as matched "pseudo" pairs (trade/tray and port/peach). These real word conditions were accompanied by matched sets of non-words (e.g. nugged/nug). Patients' latencies to the regular past tense real word-pairs were consistently slower than in any other condition. To test for a general phonological processing deficit, we conducted several tests of phonological processing ability. The results show that the patients had a range of difficulties in phonological processing, from very mild to severe. This did not correlate with their performance on the speeded judgement task. We interpret this pattern of results as support for a specialised morpho-phonological processing mechanism which can be dissociated from other phonological processes and which is used directly in the processing of the regular past tense in a dual-mechanism system. PMID- 11931920 TI - Perceptual learning in amnesia. AB - Evidence from experiments on perceptual learning, accumulated during the last few years, increasingly indicates that the relative 'front end' parts of the visual system are more plastic even in adults than was previously expected. Hence, it might be possible that perceptual learning is similar in several respects to procedural learning and may be achieved even without (declarative) memory traces present. Results on six patients suffering from global amnesia due to damage to hippocampal-diencephalic systems demonstrate, for the first time, that at least some amnesic patients are able to significantly improve performance in a visual hyperacuity task as a result of training, showing improvement as good as the observers in the control group. This result corroborates the notion of a relatively 'front end' location of at least some forms of perceptual learning. PMID- 11931921 TI - Local-global processing in Alzheimer's disease: an examination of interference, inhibition and priming. AB - Impairments of memory, praxis, gnosis, language and executive functioning are well documented in Alzheimer's disease (AD). Functions, such as attention, however, have only recently been systematically investigated. We used Navon-type stimuli (large "global" digits composed of smaller "local" digits) to assess 12 AD participants' plus age-matched controls' ability to focus and alter the scale of their spatial attention. In the first experiment, participants responded to either the global or local characters within a block, ignoring characters at the other spatial scale. Healthy young adults (n=12) demonstrated the normal 'global precedence' effect on this task. In contrast, participants with AD and their age matched controls were significantly faster on the local task than on the global task, suggesting in these groups a 'local precedence' effect. This consisted of both a local advantage and a local-on-global interference effect. In a second experiment, participants searched for designated targets which occurred unpredictably at either the local or global spatial scale. Participants with AD were significantly slower and more error-prone than older controls. In addition, participants with AD showed a greater cost in reaction time (RT) when required to switch spatial scales on consecutive trials, compared to no switch responses at the same spatial scale on consecutive trials. Thus, AD may impair the ability to process global figures, due perhaps to involvement of posterior parietal areas. Further, participants with AD were poor at inhibiting irrelevant stimuli and at inhibiting attentional allocation to an irrelevant spatial scale, which may relate to prefrontal pathology. PMID- 11931922 TI - Impaired fear conditioning in Alzheimer's disease. AB - Classical conditioning of the fear response is a basic form of nondeclarative (nonconscious) memory that mediates both normal and pathological responses to aversive stimuli. Because fear conditioning critically depends on the amygdala, a medial temporal lobe structure that frequently undergoes significant pathological changes early in the course of Alzheimer's disease (AD), we hypothesized that fear conditioning would be impaired in patients with mild to moderate AD. We examined simple classical fear conditioning in a group of 10 patients with probable AD and 14 demographically matched, neurologically intact elderly controls. During conditioning, one stimulus (e.g. a green rectangle, the conditioned stimulus (CS+)), was paired with an aversive stimulus (a loud noise, the unconditioned stimulus (US)) using a partial reinforcement conditioning schedule. The opponent color (e.g. red rectangle), the CS-, was never paired with the US. The elderly controls acquired robust fear responses as demonstrated by their differential skin-conductance responses to the CS+ and CS-. In contrast, the AD group showed a marked impairment in conditioning, failing to exhibit significant conditioned fear responses. This failure to acquire conditioned responses could not be attributed to diminished responding by patients, relative to controls, to the aversive US. The results indicate that fear conditioning, an amygdala-dependent form of memory, is impaired in AD. These findings complement previous reports of impairments in declarative emotional memory in AD by demonstrating that a basic form of nondeclarative emotional memory is also impaired in AD. PMID- 11931923 TI - Investigation of the single case in neuropsychology: confidence limits on the abnormality of test scores and test score differences. AB - Neuropsychologists often need to estimate the abnormality of an individual patient's test score, or test score discrepancies, when the normative or control sample against which the patient is compared is modest in size. Crawford and Howell [The Clinical Neuropsychologist 12 (1998) 482] and Crawford et al. [Journal of Clinical and Experimental Neuropsychology 20 (1998) 898] presented methods for obtaining point estimates of the abnormality of test scores and test score discrepancies in this situation. In the present study, we extend this work by developing methods of setting confidence limits on the estimates of abnormality. Although these limits can be used with data from normative or control samples of any size, they will be most useful when the sample sizes are modest. We also develop a method for obtaining point estimates and confidence limits on the abnormality of a discrepancy between a patient's mean score on k tests and a test entering into that mean. Computer programs that implement the formulae for the confidence limits (and point estimates) are described and made available. PMID- 11931924 TI - Motor control and cerebral hemispheric specialization in highly qualified judo wrestlers. AB - With the purpose of investigating motor and cognitive lateralization profiles associated with long-term motor training, we investigated differences in hemispheric specialization between proficient judo sportsmen and controls through the assessment of a number of handedness and footedness items including postural preferences as well as dichotic listening and lateralized visual field tests. Our data show that: (1) the different handedness and footedness items did differently relate to each other within the athlete and control groups as revealed by a principle component analysis (PCA); (2) stand side correlated differently to these motor profile factors in athletes and controls; (3) athletes preferred more frequently to perform certain movements with the left hand than controls, although overall right-handed; (4) this was especially true for athletes which proved to be most proficient/skilled; and (5) in a lateralized verbal listening task and a lateralized visual field task athletes revealed enhanced right hemispheric involvement relative to controls. Our results suggest that during motor and postural skill acquisitions (long-term judo training) lateral preferences are modified, probably due to neuroplasticity. Moreover, the present findings support the multidimensional view of handedness by Steenhuis and Bryden [Cortex 25 (1989) 289] and the notion of a right-hemispheric "praxis system" involved in skilled action routines within peripersonal space [Brain and Cognition 23 (1993) 181]. PMID- 11931925 TI - Naturalistic action impairments in dementia. AB - Naturalistic actions are everyday tasks (e.g. cooking) that require one to use multiple objects and sequence steps to achieve a goal. Naturalistic action impairment has been attributed to executive dysfunction [Higher cortical functions in man. New York: Basic Books, 1966], semantic knowledge degradation [Brain 111 (1988) 1173], and, more recently, general limitations in cognitive resources [Neuropsychology 12 (1998) 13]. Action impairments were explored in 51 dementia participants with the short form of the multi-level action test (MLAT S). A clinical neuropsychological test protocol was also administered. Regression analyses including measures of executive functioning, semantic knowledge, and global cognitive functioning showed that global cognitive functioning was the best predictor of MLAT-S errors. Furthermore, task demands significantly influenced the type and frequency of errors, and dementia participants showed a pattern of errors similar to that reported in other clinical populations [Cognitive Neuropsychology 15 (1998) 617; Neuropsychologia 37 (1999) 51; Neuropsychology 12 (1998) 13]. Taken together, the present findings are inconsistent with semantic and executive accounts, but support the limited capacity resource theory of naturalistic action impairment. PMID- 11931926 TI - Lateralization of speech production using verbal/manual dual tasks: meta-analysis of sex differences and practice effects. AB - The present paper reviews the findings of 30 years of verbal/manual dual task studies, the method most commonly used to assess lateralization of speech production in non-clinical samples. Meta-analysis of 64 results revealed that both the type of manual task used and the nature of practice that is given influence the size of the laterality effect. A meta-analysis of 36 results examining the effect size of sex differences in estimates of lateralization of speech production indicated that males appear to show slightly larger laterality effects than females. PMID- 11931927 TI - Anterior cingulate cortex and the Stroop task: neuropsychological evidence for topographic specificity. AB - Neuroimaging studies have implicated the anterior cingulate cortex (ACC) in many aspects of attention and cognition. Major theories of ACC function have proposed a role in conflict monitoring, executive control, response selection, and general arousal. Although the ACC is often treated as a unitary structure, extensive evidence suggests it exhibits anatomical and functional specificity. ACC activity during the Stroop color word interference task has been of particular interest. The purpose of the present study was to determine whether two different ACC subregions are necessary for intact color naming performance in the Stroop task. One experiment utilized blocked trial and mixed trial designs to emulate neuroimaging studies and to compare interference and facilitation effects, respectively. A third variant manipulated the probabilities of congruent and incongruent trials to alter levels of interference and cognitive control, or engagement of strategic processes, on a block by block basis. Two patients with focal lesions in either right mid-caudal (patient D.L.) or left rostral to mid dorsal ACC (patient R.N.) exhibited distinctive performance profiles in these three versions of the Stroop task, providing further support for topographic specificity of function within the human ACC. Contrary to predictions from some neuroimaging experiments, damage to right mid-caudal ACC was associated with normal levels of interference and accurate performance on incongruent trials. Instead, D.L. showed reduced levels of facilitation relative to controls. Further, interference was not modulated by the probability manipulation in D.L., suggesting equivalently high levels of cognitive control in both conditions. Conversely, damage to left mid-dorsal ACC resulted in consistently lower accuracy on incongruent trials, indicating deficits in maintaining task set and inhibiting the automatic response. These results can help to constrain interpretations of ACC activations in functional imaging experiments of the Stroop task. PMID- 11931928 TI - Brain activity related to integrative processes in visual object recognition: bottom-up integration and the modulatory influence of stored knowledge. AB - We report evidence from a PET activation study that the inferior occipital gyri (likely to include area V2) and the posterior parts of the fusiform and inferior temporal gyri are involved in the integration of visual elements into perceptual wholes (single objects). Of these areas, the fusiform and inferior temporal gyri were more activated by tasks with recognizable stimuli than by tasks with unrecognizable stimuli. We propose that the posterior parts of the fusiform and inferior temporal gyri, compared with the inferior occipital gyri, are involved in higher level integration, due to the involvement of re-entrant activation from stored structural knowledge. Evidence in favor of this interpretation comes from the additional finding that activation of the anterior part of the left fusiform gyrus and a more anterior part of the right inferior temporal gyrus, areas previously associated with access to stored structural knowledge, was found with recognizable stimuli, but not with unrecognizable stimuli. This latter finding also indicates: (i) that subjects may not refrain from (automatically) identifying objects even if they only have to attend to the objects' global shape, and (ii) that perceptual and memorial processes can be dissociated on both functional and anatomical grounds. No evidence was obtained for the involvement of the parietal lobes in the integration of single objects. PMID- 11931929 TI - On the role of the SMA in the discrete sequence production task: a TMS study. Transcranial Magnetic Stimulation. AB - Participants practiced two discrete six-key sequences for a total of 420 trials. The 1 x 6 sequence had a unique order of key presses while the 2 x 3 sequence involved repetition of a three-key segment. Both sequences showed a long interkey interval halfway the sequence indicating hierarchical sequence control in that not only the 2 x 3 but also the 1 x 6 sequence was executed as two successive motor chunks. Besides, the second part of both sequences was executed faster than the first part. This supports the earlier notion of a motor processor executing the elements of familiar motor chunks and a cognitive processor triggering either these motor chunks or individual sequence elements. Low-frequency, off-line transcranial magnetic stimulation (TMS) of the supplementary motor area (SMA) counteracted normal improvement with practice of key presses at all sequence positions. Together, these results are in line with the notion that with moderate practice, the SMA executes short sequence fragments that are concatenated by other brain structures. PMID- 11931930 TI - Verbal fluency in Huntington's disease: a longitudinal analysis of phonemic and semantic clustering and switching. AB - Two underlying components of verbal fluency tasks have been identified as clustering (the ability to generate successive words within a sub-category) and switching (the ability to shift from one sub-category to another). Selective impairment of switching ability occurs in patients with frontostriatal pathology, whilst clustering ability is compromised with temporal lobe dysfunction. Letter fluency tasks have been shown to be especially sensitive to frontostriatal deficits, whereas, category fluency tasks tend to be compromised by temporal lobe pathology. This study examined two types of verbal fluency task (letter fluency and category fluency) using two levels of analysis (phonemic and semantic) for clustering and switching measures. The performance of 21 frontostriatally compromised Huntington's disease (HD) patients was followed over an average of 3.5 annual follow-up visits. HD patients showed a significant reduction of correctly generated words over time, together with a significant increase in word repetitions. Phonemic switching decreased significantly over time for both letter and category fluency. Semantic switching, however, remained stable over time for both verbal fluency tasks. Clustering (both semantic and phonemic) likewise remained stable and did not vary longitudinally for either letter or category fluency. Hence, phonemic switching alone drove verbal fluency performance and this selective impairment can be explained by the progressive involvement of frontostriatal circuitry in the natural progression of HD. PMID- 11931931 TI - Cerebellar morphology in developmental dyslexia. AB - Recent evidence has suggested cerebellar anomalies in developmental dyslexia. Therefore, we investigated cerebellar morphology in subjects with documented reading disabilities. We obtained T1-weighted magnetic resonance images in the coronal and sagittal planes from 11 males with prior histories of developmental dyslexia, and nine similarly-aged male controls. Proton magnetic resonance spectra (TE=136ms, TR=2.4s) were obtained bilaterally in the cerebellum. Phonological decoding skill was measured using non-word reading. Handedness was assessed using both the Annett questionnaire of hand preference and Annett's peg moving task. Cerebellar symmetry was observed in the dyslexics but there was significant asymmetry (right grey matter>left grey matter) in controls. The interpretation of these results depended whether a motor- or questionnaire-based method was used to determine handedness. The degree of cerebellar symmetry was correlated with the severity of dyslexics' phonological decoding deficit. Those with more symmetric cerebella made more errors on a nonsense word reading measure of phonological decoding ability. Left cerebellar metabolite ratios were shown to correlate significantly with the degree of cerebellar asymmetry (P<0.05) in controls. This relationship was absent in developmental dyslexics. Cerebellar morphology reflects the higher degree of symmetry found previously in the temporal and parietal cortex of dyslexics. The relationship of cerebellar asymmetry to phonological decoding ability and handedness, together with our previous finding of altered metabolite ratios in the cerebellum of dyslexics, lead us to suggest that there are alterations in the neurological organisation of the cerebellum which relate to phonological decoding skills, in addition to motor skills and handedness. PMID- 11931933 TI - Interhemispheric neural summation in the split brain with symmetrical and asymmetrical displays. AB - The present study, investigates interhemispheric integration in the split brain. Four split-brained, two acallosal and 14 normal subjects carried out a simple reaction time task in which they responded to stimuli presented either singly in the left visual field, singly in the right visual field, or simultaneously in both visual fields. Stimuli were white against a black background and bilateral stimuli were either symmetrical or asymmetrical around the central vertical meridian. For unilateral stimuli, the difference in response time (RT) between crossed and uncrossed hand-field combinations (crossed-uncrossed difference, or CUD) measured interhemispheric transfer time. RTs to bilateral and unilateral stimulus displays were compared to provide a measure of redundancy gain (RG). Normal subjects exhibited small CUDs and RGs. Split-brained and acallosal subjects were found to have much longer CUDs, and to show enhanced RGs which could not be explained by a probability (race) model, implying subcortical neural summation. This summation did not depend on the preservation of symmetry, suggesting that it may not occur at the retinotopically organized superior colliculus, but at another site such as the pons or reticular formation. PMID- 11931932 TI - Replication of a premenstrual decrease in right-ear advantage on language-related dichotic listening tests of cerebral laterality. AB - Five fused dichotic word tasks measured perceptual asymmetry in 30 women at 4 weekly intervals. The five tasks varied according to whether the stimuli presented were word-word pairs or nonsense word-pairs, or whether they consisted of neutral words paired with positive emotion-evoking words (e.g. hug-tug), neutral words paired with negative emotion-evoking words (e.g. till-kill), or neutral words paired with neutral words (e.g. bean-dean). Overall right-ear advantage (REA) decreased in the premenstrual week relative to the postmenstrual week, replicating previous results using identical measures. In addition, REA scores were similar at menstrual, postmenstrual and midcycle weeks. Additional data from 12 men suggests sex differences in task performance were small or non existent. In both women and men, there were no effects of repeated testing on REA, but emotional proclivity indices, defined as the tendency to recall words of positive or negative affective tone, increased across the four test sessions. As found previously, there was a trend for women to hear fewer positive words during the premenstrual week. These data are consistent with other research suggesting that a progesterone-mediated decrease in functional asymmetry occurs in the luteal phase. Future research manipulating task demands (e.g. memory load) or the affective valence of the stimuli may be useful in understanding the observed changes in hemispheric advantage. PMID- 11931934 TI - Error correction strategies for motor behavior after unilateral brain damage: short-term motor learning processes. AB - In order to identify the mechanisms underlying motor impairments and motor learning following stroke-related brain damage, we analyzed correction strategies used by hemiparetic individuals to produce precise elbow flexion movements of the paretic arm and compared them to those of healthy individuals. Participants made rapid elbow flexion movements to a 6 degrees wide target and were instructed to correct movement errors as quickly as possible when a spring-like load was unexpectedly introduced. Angular positions and torques before correction were used to identify error patterns. Results showed that participants with mild hemiparesis minimized movement errors within three trials, as did healthy participants. In contrast, severely affected individuals needed more trials to diminish errors and their movements were inconsistent. Participants with a moderate motor disability used both typical and atypical correction strategies. The differences in correction behaviors likely reflect deficits in arm motor function (r=0.79) and executive function (r=0.58) rather than levels of intellectual function (IQ ratings). Results indicate that the deficits that individuals with stroke experience when adapting their movements to changed load conditions may be due to difficulty in rapidly integrating visual and proprioceptive information. Deficits in executive function could also contribute to problems in producing accurate and consistent movements from trial to trial. Taken together, these results imply that all hemiparetic individuals would not benefit equally from the same motor re-training approaches. PMID- 11931935 TI - Dissociating habit and recollection: evidence from Parkinson's disease, amnesia and focal lesion patients. AB - We investigated the role played by the striatum and the medial temporal lobes (MTL) in memory performance by testing patients with Parkinson's disease (PD) and amnesia with Hay and Jacoby's habit-learning task [Journal of Experimental Psychology: Learning, Memory and Cognition 22 (1996) 1323]. Using equations from Jacoby's process-dissociation procedure [Journal of Memory and Language 30 (1991) 513], we were able to separate out the contribution of habit (automatic memory) and recollection (intentional memory) to performance within a single probability learning paradigm. Amnesics showed the expected dissociation of impaired recollection and intact habit, highlighting the important role of the MTL in recollective processing. Mild PD patients did not perform differently than matched controls for habit or recollection, however, moderate PD patients were impaired in their ability to rely on habit and in their ability to recollect specific information. The performance of focal lesion patients further supported the interpretation that PD patients have a significant deficit in automatic, habit-learning due to striatal dysfunction while their deficit in recollection may arise from impoverished frontal lobe contributions. PMID- 11931936 TI - Material-specific deficits in "remembering" in patients with unilateral temporal lobe epilepsy and excisions. AB - A growing body of research suggests that "remembering" and "knowing" may be dissociable aspects of recognition memory. Cognitive theorists have argued that the former reflects conceptual processing and is based on distinctive memory traces whereas the latter is associated with perceptual analysis and reflects fluency of processing. Here, we investigate whether this framework can account for memory deficits observed following right or left temporal lobe damage, as suggested by Blaxton and Theodore [Brain and Cognition 35 (1997) 5]. Recognition memory for faces and words was examined in patients with unilateral temporal lobe epilepsy or excisions (TLE) and controls using the "remember-know" recognition paradigm. Participants studied items under conditions designed to enhance either conceptual processing or focus their attention on superficial aspects of the items. For controls, there was an increase in "remember" responses following conceptually-based encoding for both words and faces. This enhancement was eliminated in patients with left TLE for words and those with right TLE showed a diminished effect for faces. This pattern indicates an impairment in the ability to benefit from conceptual encoding that is specific to material preferentially processed by the damaged hemisphere. Furthermore, this effect was only observed for "remember" responses, which are based on the participants' ability to recollect specific contextual aspects of the original study experience. These data can be interpreted in relation to current theories of hippocampal function, which emphasize the critical role played by the hippocampus in relational memory formation and retrieval. We offer this as a novel interpretation of the "remember know" literature. PMID- 11931937 TI - Further delineation of the executive deficit in males with fragile-X syndrome. AB - This paper presents a detailed analysis of one aspect of performance by young males with fragile-X syndrome (FMR-1 full mutation) who were assessed on a computerised visual search task as part of a larger study examining aspects of attention [Neuropsychologia 38 (2000) 1261]. They were matched on chronological and mental age to 25 boys with Down's syndrome (trisomy 21) and on mental age to 50 mainstream school boys (controls). The controls were further divided into those matched on "poor attention" to the fragile-X boys and a "good" attention group, as rated by the comprehensive teacher rating scale (ACTeRS) questionnaire. Both fragile-X and Down's syndrome boys made significantly more repeated responses on targets (but a lower proportion of errors based on confusion of shape) than the two control groups and these differences were stronger in the fragile-X group. In the single target condition, search was for a single type of target throughout. In the dual target condition, participants were required to alternate between two different targets. Fragile-X boys showed significantly greater inability than Down's syndrome and normal boys to switch attention between targets and both learning-disabled groups were inferior to the control groups. Thus, both learning-disabled groups displayed a weakness in inhibiting repetition and in switching attention from one type of target to another and the impairments were more acute in fragile-X boys. The results provide further support for an attention deficit in this population at higher levels of attention control/executive functioning that involve switching visual attention and inhibiting repetitious behaviour. PMID- 11931938 TI - Left hand movements and right hemisphere activation in unilateral spatial neglect: a test of the interhemispheric imbalance hypothesis. AB - The aim of the present study was to check one of the main assumptions of the interhemispheric imbalance hypothesis, namely, the prediction that the severity of neglect should be reduced by conditions activating the right hemisphere. To achieve this, a group of neglect patients was studied using a slightly modified version of the limb activation technique. The (verbal or visuo-spatial) nature of the stimuli to be processed by the patient and the (left or right) side of space where the left hand moved were considered as the critical variables to check the interhemispheric imbalance hypothesis. Three traditional and one new methods were used to measure changes induced in the severity of neglect by the material to be processed or by the side of space where the left hand moved. The traditional methods, all based on counting omissions, consisted of measuring: (a) the overall number of omissions; (b) the number of omissions made on the left half sheet; or (c) the difference between the omissions made on the left and right sides of the sheet. The new index, based on the notion of the 'attentional field' and defined as the spatial distribution of stimuli detected by the patient, was operationally measured by computing the distance between each stimulus crossed out by the patient and the right margin of the sheet. The study was conducted by rating the severity of neglect in 42 cancellation sheets which had used, respectively letters (N=21) and small geometric figures (N=21) as targets. The two sets of cancellation sheets were obtained from seven neglect patients during a limb activation task requiring the cancellation of a given target in three different conditions: (a) baseline; (b) active movements of the left hand in the left half space; (c) active movements of the left hand in the right half space. Results were at variance with the predictions based on Kinsbourne's model, since the verbal or visual spatial nature of the material to be processed did not influence the severity of unilateral spatial neglect (USN) and since left hand movements produced a significant reduction in the severity of neglect only when these movements were made on the left side of space. PMID- 11931939 TI - Probabilistic classification learning in Tourette syndrome. AB - Tourette syndrome (TS) is characterised by stereotyped involuntary movements, called tics. Some evidence suggests that structural and functional abnormalities of the basal ganglia may explain these motor symptoms. In this study, the probabilistic classification learning (PCL) test was used to evaluate basal ganglia functions in 10 children with less severe tics (Yale Global Tic Severity Scale (YGTSS) scores<30) and in 10 children with more severe symptoms (YGTSS score>30). In the PCL task, participants are asked to decide whether different combinations of four geometric forms (cues) predict rainy or sunny weather. Each cue is probabilistically related to a weather outcome, and feedback is provided after each decision. After completion of the probabilistic stimulus-response learning procedure, subjects received a transfer test to assess explicit knowledge about the cues. The children with TS exhibited impaired learning in the PCL task in comparison with the 20 healthy control subjects. This impairment was more pronounced in the TS patients with severe symptoms, and there was a significant negative relationship between the final classification performance and the YGTSS scores. The patients showed normal learning in the transfer test. These results suggest that the neostriatal habit learning system, which may play a central role in the acquisition of probabilistic associations, is dysfunctional in TS, especially in the case of more severe motor symptoms. The classification performance and the severity of tics were independent of the explicit knowledge obtained during the test. PMID- 11931940 TI - Visually- and motor-based knowledge of letters: evidence from a pure alexic patient. AB - We describe a patient, VSB, whose reading was impaired as a consequence of a left temporal-parietal lesion, whereas writing was relatively preserved. At variance with other pure alexic patients described in the literature, VSB claimed to have become unable to mentally visualise letters and words. Indeed, his performance on a series of tests tapping visual mental imagery for orthographic material was severely impaired. However, performance on the same tests was dramatically ameliorated by allowing VSB to trace each item with his finger. Visual mental imagery for non-orthographic items was comparatively spared. The pattern of dissociation shown by VSB between impaired visual mental imagery and relatively preserved motor-based knowledge for orthographic material lends support to the view that separate codes, respectively based on visual appearance and on motor engrams, may be used to access knowledge of the visual form of letters and words. PMID- 11931941 TI - Line bisection in normal adults: direction of attentional bias for near and far space. AB - The direction of attentional bias in forty normal adults was assessed using a computer generated line bisection task. A 4 (viewing distance)x4 (line length)x2 (cursor starting position) repeated measures factorial design was employed. As predicted, differences in bisection performance as a function of viewing distance were observed. The findings confirmed that scanning direction (contingent upon the starting position of the cursor), but not line length, significantly modulated this effect. The direction of bias across near and far space was further clarified yielding a progressive shift from a leftward bias in near space to a rightward bias in far space. A significant interaction of distance, line length and starting position revealed differential effects for left and right starting positions as a function of viewing distance and line length. More specifically, a leftward start witnessed deviations shifting progressively from left-to-right as distance and line length increased though no comparable pattern was observed for rightward starts. The results provide important behavioural support for the suggestion that dissociated neural systems may be responsible for attending and acting in near and far space and that other lateralised functions (such as scanning strategies) can influence hemispheric activation. The findings have relevant theoretical implications as well as important implications for the clinical assessment of unilateral neglect using a standard line bisection task, both of which are discussed. PMID- 11931942 TI - Laterality effects on motor awareness. AB - The ability to perceive self-produced movements and to correctly attribute an action to its proper agent is a natural task and a basic requirement to human social communication. Recent experiments suggest that this apparently simple phenomenon is related to the mechanisms of motor production, raising the question whether recognition of self-produced movement is affected by asymmetries similar to those present in motor skills. In this study, right- and left-handed subjects decided whether a moving hand presented on a screen was the image of their own hand or of that of another person. Two experimental conditions were tested: either subjects saw their own moving hand (subject condition) or they were shown the experimenter's hand pantomiming their movement (experimenter condition); a glove masked morphological differences between the two hands. Verbal responses and response times were analysed. Results showed that all subjects were more accurate in recognising their preferred hand with respect to their non-preferred hand. Right-handers responded faster than left-handers did, the latter group being especially slowed down in the experimenter condition. On the contrary, in the right-handers group, response times did not differ among conditions. The present data indicate that the ability to recognise self-generated movements is affected by motor dominance, thus suggesting that conscious knowledge of self produced movements is closely related to the motor system. PMID- 11931943 TI - Categorisation of 'perceptual' and 'premotor' neglect patients across different tasks: is there strong evidence for a dichotomy? AB - The aim of the presented studies was to investigate whether classifications of neglect patients into perceptual (i.e. identifying a patient as suffering from mainly attentional/space representation deficits) and premotor (judging the main impairment to be related towards actions into contralesional space) categories is consistent across different line bisection assessment techniques that have, in the past, been designed to tease these potentially overlapping aspects of hemispatial neglect apart. Twelve patients with hemispatial neglect and three control groups were tested with the Overhead Task, adapted from Nico [Neuropsychologia 34 (1996) 471] in which patients were asked to bisect lines that were mirror reversed, the Pulley Device Technique, adapted from Bisiach et al. [Neurology 40 (1990) 1278] in which they had to perform a movement opposite to the direction of the transaction mark that bisected the line and the Landmark Test, adapted from Milner et al. [Neuropsychologia 30 (1992) 515] in which they had to manually point to the half of a centrally pre-bisected line that, to them, appeared shorter. The specific question was whether these three tasks would categorise the same set of patients in the same way?Most patients could be classified into either the premotor or perceptual category in each task, but no consistent categorisation emerged across the three techniques. Just 1 out of the 12 patients, was consistently classified across all three tasks. It seemed that despite the fact that all tasks essentially required a line bisection response, the perceptual and motor differences between the tasks were still great enough to result in inconsistent classifications. The Landmark Task classified the majority of patients into the perceptual neglect category, while the Overhead and Pulley Device Techniques tended to identify more patients as suffering from a premotor deficit (albeit not the same set of patients). PMID- 11931944 TI - Audiovisual speech perception in Williams syndrome. AB - People with the genetic disorder of Williams syndrome (WS) show an anomalous cognitive profile, wherein some purely verbal and social communicative abilities are relatively proficient, while visuo-spatial skills can be extremely impaired. Face processing, while apparently relatively spared among visuo-spatial skills, can show deficits suggesting developmental immaturity. In this context, the exploration of visual and audiovisual speech perception in WS is of interest. A new test based on tokens from a single natural English speaker of the form /(inverted v)ba:/, /(inverted v) va:/, /(inverted v)(theta)a:/, /(inverted v)da:/ and /(inverted v)ga:/, digitally manipulated and presented in unimodal (vision alone, audition alone) and audiovisual conditions, was presented for participants to identify each token. Compared with age-matched controls, WS participants were impaired at visual but not auditory identification, and in audiovisual testing showed correspondingly reduced effects of vision on report of auditory token identity. Audiovisual integration was nevertheless demonstrable in WS. Speech reading may require skills which do not reach age-appropriate levels in WS, despite their age-appropriate (auditory) phonological abilities. PMID- 11931945 TI - Dissociable processing of temporal structure in repetitive eye-hand movements in Parkinson's disease. AB - Movement takes place in multiple dimensions, including the temporal dimension, which itself may be made up of dissociable aspects. From this perspective, the present research tests three hypotheses: first, that the generation of regular, repetitive movement frequency is neurophysiologically dissociable from the generation of appropriate phase relations, or latencies, of such movements with respect to external stimuli; second, that the frontostriatal system is critically involved in the control of latency and not frequency and third that while the control of latency is closely linked to the effector motor system (e.g. eye, hand, etc.) the control of movement frequency is a more centralized function. These three hypotheses were investigated in nine Parkinson's disease (PD) patients with asymmetric akinetic-rigid syndrome and in nine age-matched control subjects in the context of repetitive eye-hand movements generated in response to regularly displaced visual stimuli and then the continuation of these movements in the immediate absence of the stimuli. PD patients demonstrated increased latencies for pointing movements, particularly with the affected hand, while their ability to follow and then reproduce the movement frequency remained largely intact. Interestingly, saccade latency was improved for controls and PD patients when pointing with the Less-affected hand and impaired with the More affected hand. In contrast, saccade frequency was unaffected in these pointing conditions. These results support the hypothesis that movement frequency and latency controls are dissociable, with the frontostriatal system playing an important role in latency but not frequency control. The fact that pointing and saccade latency displayed a hand-effect, while movement frequency did not, also tends to support the hypothesis that latency control is linked to specific motor systems, including their interaction, whereas frequency control is more centralized. PMID- 11931946 TI - Oral fluency and narrative production in children with Turner's syndrome. AB - Despite good verbal skills, children with Turner's syndrome (TS) have been found to perform poorly on oral fluency tasks. Explanations for this impairment were explored in a study of 9-12-year-old children with TS. The children with TS were found, as expected, to have impaired levels of retrieval on initial letter fluency tasks, in comparison to their peers. Children with TS were also found to have receptive vocabularies which were significantly better than controls, indicating that the weak performance on oral fluency cannot be attributed to lower vocabulary levels. Naming skills were normal for children with TS, indicating that oral fluency scores cannot be attributed to generalised problems with lexical access. Analysis of the content of fluency performance indicated clusters of words within sub-categories which were of normal size but there were fewer switches between clusters than for controls and fewer returns to previously successful clusters. Additionally, a significantly larger proportion of the words generated were of low frequency (e.g. Assyrian, antediluvian) and the children with TS did not thereby show a normal frequency effect in word retrieval. One explanation is that these reflect abnormal action of executive language retrieval processes. Within narrative production, picture description was normal. However, in relation to narrative tasks, hypothesised to place greater executive demands, there was impairment which was significantly greater for narratives of yesterday than narratives of organising a party. This impairment within the narrative generation tasks could be attributable to a selective impairment within executive retrieval skills, or could represent an impairment in episodic memory or the executive processes involved in its retrieval. PMID- 11931947 TI - Neural correlates of memory for object identity and object location: effects of aging. AB - The purpose of this study was to investigate the effects of aging on memory for object identity and object location to determine whether aging affects both posterior neocortical areas that are domain-specific and other brain regions, such as pre-frontal cortex, that are involved in encoding and retrieval regardless of the information that is processed (domain-general). We used positron emission tomography (PET) to measure changes in regional cerebral blood flow (rCBF) in younger and older participants while they were engaged in encoding and retrieving information about object identity and object location. Compared to young adults, older adults showed decreased activation in domain-specific regions of inferior parietal and inferior temporal cortex while engaged in processing (encoding and retrieving) information about object location and object identity, respectively. This decreased specificity in the older adults was accompanied by greater domain-general activation in right prefrontal and premotor cortex during perceptual encoding than during retrieval. Conversely, the younger participants showed greater domain-general activation in right extrastriate cortex (Brodmann area (BA) 18) during retrieval. Moreover, we found that medial temporal and frontal lobes were synergistically activated in younger adults but not in older adults. The pattern of decreased specificity of activation in posterior neocortex with greater activation in anterior neocortex suggests that, with age, compensatory domain-general mechanisms in anterior neocortex are recruited to mitigate altered domain-specific processes. Thus, the results of the present study indicate that the relation between the presumed integrity of various structures, such as the hippocampus, prefrontal cortex, and posterior neocortex, and their pattern of activation, is a complex one that is influenced by age, by the perceptual and cognitive demands of the task and their interaction. PMID- 11931948 TI - Spatial behaviour is driven by proximal cues even in mildly impaired Parkinson's disease. AB - This investigation addresses the question whether patients with mild to moderate Parkinson's disease (PD) show spatial deficits in real-life settings. Therefore, a "search through"-locomotor task incorporating basic features of both the radial arm maze and the Morris water maze paradigms was used. The participants had to find and remember five out of twenty hidden locations within a completely controlled environment. Different spatial memory errors and inter-response intervals were recorded automatically. Fourteen patients with idiopathic PD and fourteen healthy controls matched for age, sex, handedness, and education were investigated. Patients and controls were widely comparable with respect to intelligence, verbal memory, and executive functions. Results show that performance deteriorated in about half of the patients if the starting position was moved by 90 degrees and the proximal cues were deleted simultaneously. This deficit was systematically related to specific measures of attention. Moreover, patients were less able to update their locomotor strategies towards a more effective strategy. Results are discussed with respect to the patient's inability to generate rules which can be flexibly used in changing environments, especially if proximal cues are removed. It is concluded that deficits in spatial behaviour can be elicited even in near-to-real-life situations if appropriate testing procedures are used. PMID- 11931949 TI - Novelty-related activation within the medial temporal lobes. AB - Functional magnetic resonance imaging (fMRI) was used to examine whether (1) verbal associative encoding activates the medial temporal lobes (MTL) and related regions more than non-associative encoding, (2) verbal associative novelty is related to enhanced MTL activation, and (3) verbal item novelty is related to enhanced MTL activation and, if so, whether these activations are in different or overlapping sites. No increase in MTL activation was found during verbal associative encoding relative to non-associative encoding, although associative encoding was related to a relative increase in activation in the posterior cingulate cortex. In contrast, verbal associative novelty was found to activate the MTL and posterior cingulate cortex. Verbal item novelty did not significantly activate any brain region. The verbal associative novelty-related effect occurred despite subjects having little awareness of associative novelty. The verbal associative novelty-related activation in the MTL may be related either to unconscious novelty detection or to a priming effect at encoding. We argue that if the priming explanation is correct then this may account for our failure to observe an associative encoding MTL activation. PMID- 11931950 TI - Lateralization of spatial-memory processes: evidence on spatial span, maze learning, and memory for object locations. AB - Spatial memory is one of the most important cognitive functions in daily life, enabling us to locate objects in our environment or to learn a route or a path. In the present study, we elaborated on the hypothesis that human spatial memory consists of multiple sub-processes, relying on different brain structures. Therefore, 50 patients with an ischemic stroke and 40 healthy participants underwent tests measuring spatial span and maze learning. By means of a computer paradigm the following aspects of memory for object locations were assessed: (1) object location binding; (2) positional memory; (3) a combination of these two aspects. The results clearly showed a double dissociation: the group of patients with an infarct in the left hemisphere (LH) was impaired on object location binding, whereas the group with an infarct in the right hemisphere (RH) was impaired on positional memory. Lesions in the RH resulted also in impairments on maze learning. Moreover, patients with lesions in the posterior part of the parietal or the occipital lobe performed especially worse on spatial-memory tasks. These findings extend the theoretical framework of categorical versus coordinate spatial processing in the human brain and corroborate previous findings on selective aspects of memory for object locations. PMID- 11931951 TI - Grey scales uncover similar attentional effects in homonymous hemianopia and visual hemi-neglect. AB - Multi-component models of visual hemi-neglect have postulated that visual hemi neglect is characterised by various attentional deficits. A grey scales task has been developed to quantify the early, automatic, (perhaps obligatory) ipsilesional orienting of visual attention, frequently assumed as the first of these attentional deficits. Explanations for this attentional imbalance are up until now mainly formulated in terms of right hemisphere activation. This lateral attentional bias has also been demonstrated in controls, in whom it is expressed as a leftward perceptual asymmetry. We reproduced previous literature findings on a grey scales task, considering controls and neglect patients. Three patients with neglect showed an extreme ipsilesional lateral bias. This bias did not change during or after cognitive rehabilitation. Additionally, we presented this grey scale task to 32 patients with left- and right-sided homonymous hemianopia (HP). HP is the loss of sight in one visual hemi-field. The HH patients had no clinical signs of impaired lateralised attention. Results revealed that HH patients showed a similar ipsilesional bias, albeit to a lesser degree than in neglect. Left-sided HH patients presented a quantitatively similar, but qualitatively opposite bias than the right-sided HH patients. We suggest that sensory effects can be an alternative source of attentional imbalance, which can interact with the previously proposed (right) hemispheric effects. This suggests that the perceptual asymmetry in the grey scales task is not necessarily an indicator of impaired right hemisphere attention. It rather suggests a pattern of functional cerebral asymmetry, which can also be caused by asymmetric sensory input. PMID- 11931952 TI - Drawing and identifying objects in relation to semantic category and handedness. AB - We investigated whether the directionality of different objects and participants' handedness affected drawing and visual object identification performance. In Experiment 1, 190 participants were asked to draw 246 pictures of objects belonging to different semantic categories. Results showed a preferential direction (leftward, rightward, or frontal view) for most of pictures. In Experiment 2, the directionality patterns found in Experiment 1 were verified in a visual identification task. Sixty participants were requested to identify pictures presented leftward or rightward. For some categories, the speed of responses was greater for pictures having the directionality assessed in the previous experiment. The effect of handedness emerged for two categories of objects (animals and vehicles) characterised by two properties: asymmetry and motion. Findings were discussed in relation to the Motor Image Theory on the isomorphism between motor (drawing) and visual (identification) processes. PMID- 11931953 TI - Efficiency of temporal order discrimination as an indicator of bradyphrenia in Parkinson's disease: the inspection time loop task. AB - To investigate the bradyphrenia hypothesis of Parkinson's disease (PD), 32 patients undertook an information-processing task which measured their efficiency of temporal order discrimination. Their performance was compared with 31 non-PD controls matched on age, sex, years of full-time education and pre-morbid IQ. The task was novel and designed to be sensitive to the clinical phenomenon of bradyphrenia (slowing of mental abilities), in the context of temporal order discrimination without confounding from motor ability deficits. The test (the inspection time loop task; ITloop) required judgements as to the temporal sequence of four single letters. The stimulus duration of the letters in each sequence ranged from 100 to 700ms. The PD group had a significantly lower mean score on the ITloop task than did controls (P=0.02). PD patients perform more poorly on temporal order discrimination judgements even when the task makes no motor demands. PMID- 11931954 TI - Modelling rhythmic function in a musician post-stroke. AB - The aim of this study was to model the components of rhythmic function in a case (H.J.) of acquired rhythmic disturbance. H.J. is a right-handed, amateur male musician who acquired arrhythmia in the context of a global amusia after sustaining a right temporoparietal infarct. His rhythmic disturbance was analysed in relation to three independent components using an autoregressive extension of Wing and Kristofferson's model of rhythmic timing. This revealed preserved error correction and motor implementation capacities, but a gross disturbance of H.J.'s central timing system ("cognitive clock"). It rendered him unable to generate a steady pulse, prevented adequate discrimination and reproduction of novel metrical rhythms, and partly contributed to bi-manual co-ordination difficulties in his instrumental performance. The findings are considered in relation to the essential components of the cognitive architecture of rhythmic function, and their respective cerebral lateralisation and localisation. Overall, the data suggested that the functioning of the right temporal auditory cortex is fundamental to 'keeping the beat' in music. The approach is presented as a new paradigm for future neuropsychological research examining rhythmic disturbances. PMID- 11931955 TI - Presentation and task effects on migration errors in attentional dyslexia. AB - We report a case study of a patient, FL, who shows symptoms of attentional dyslexia. He performs better at reading aloud single words than pairs of words, where he tends to make letter migration errors. These letter migrations are particularly prevalent from the word presented on the right into FL's response to the word presented on the left. Performance on the word pair reading task is improved if the two words are presented in different cases or the space between them is increased. Further, FL performs better at reading a target word if he is only required to name the initial letter of a distractor word rather than read the whole word. However, single letter distractors then produce more interference than whole words. These findings are consistent with FL having poor letter location coding when attention is diffuse [Cognitive Neuropsychol. 18 (2001) 551; Cognitive Neuropsychol. 13 (1996) 205]. In addition, representations on the left side of his attentional space are particularly weak, and so vulnerable to stimuli on the right when FL adopts a wide spatial window for the task. The data point to a pre-categorical deficit in reading in this case. PMID- 11931956 TI - Sex and spatial position effects on object location memory following intentional learning of object identities. AB - Memory for object location relative both to veridical center (left versus right visual hemispace) and to eccentricity (central versus peripheral objects) was measured in 26 males and 25 females using the Silverman and Eals Location Memory Task. A subset of participants (17 males and 13 females) also completed a measure of implicit learning, the mirror-tracing task. No sex differences were observed in memory for object identities. Further, in both sexes, memory for object locations was better for peripherally located objects than for centrally located objects. In contrast to these similarities in female and male task performance, females but not males showed better recovery of object locations in the right compared to the left visual hemispace. Moreover, memory for object locations in the right hemispace was associated with mirror-tracing performance in women but not in men. Together, these data suggest that the processing of object features and object identification in the left cerebral hemisphere may include processing of spatial information that may contribute to superior object location memory in females relative to males. PMID- 11931957 TI - Equilibrium and structural studies on metal complexes of carbohyrates and their derivatives. AB - A summary is presented of the studies of our group on metal complexes of carbohydrates (aldoses, ketoses, mono-, di- and polysaccharides) and their derivatives (aldonic, alduronic acids, polyalcohols, amino sugars, amino acid sugar adducts, AMP, ATP, etc.). The results are reported of equilibrium, electrochemical, solution and solid-state structural studies of complexes of transition metals [Cu(II), Fe(III), Ni(II), Zn(II), Co(II), Ag(I), Mn in different oxidation states and organotin(IV)]. The structural parameters (coordination number, bond distance, and Debye-Waller factor) obtained by extended X-ray absorption fine structure spectroscopic (EXAFS) spectroscopy are discussed in detail. The general rules concerning the formation and structure of such complexes are emphasized. PMID- 11931958 TI - Bisphosphonate chelating agents. Coordination ability of 1-phenyl-1 hydroxymethylene bisphosphonate towards Cu(2+) ions. AB - Potentiometric and EPR data allow for evaluation of the coordination equilibria in the Cu(2+)-bisphosphonate system. The bisphosphonic ligand was found very efficient in Cu(2+) chelation with formation of monomeric and dimeric species. Two phosphonate groups are basic binding sites for metal ion. The involvement of hydroxyl in metal ion coordination is also likely, especially when one phosphonate is protected by dimethyl ester. As the metal bound phosphonate groups are relatively bulky (six oxygens) and their negative charge above pH 4 is high (four per ligand) the equimolar species is a dominant complex at physiological pH. PMID- 11931959 TI - Synthesis, crystal structure, EPR properties, and anti-convulsant activities of binuclear and mononuclear 1,10-phenanthroline and salicylate ternary copper(II) complexes. AB - Two ternary Cu(II) complexes of 1,10-phenanthroline (phen) and singly (Hsal(-)) or dideprotonated (sal(2-)) salicylate ligands were synthesized, their X-ray crystal structure and electron paramagnetic resonance spectral characteristics determined, and evaluated for anti-convulsant activities in the maximal electroshock (MES) and Metrazol models of seizure and Rotorod toxicity. The X-ray crystal structure of [bis(1,10-phenanthroline)-mu-bis(salicylato O,O')dicopper(II)] dihydrate, 1, ([Cu(II)(2)(phen)(2)(sal)(2)].2[H(2)O]), shows it to be binuclear. This dimer consists of two centrosymmetrically related pseudo five coordinate Cu(II) atoms 3.242(2) A apart and bridged by two dideprotonated salicylate ligands. The X-ray crystal structure of [bis(1,10 phenanthroline)(salicylato)copper(II)][salicylate] monohydrate, 2, ([Cu(II)(phen)(2)(Hsal)](+)[Hsal](-)[H(2)O]), shows it to be mononuclear. This complex cation exhibits a highly irregular distorted square pyramidal geometry about the Cu(II) atom, (4+1+1*). Each salicylate is singly deprotonated and one of them is ligand bonded in an asymmetric chelating mode. EPR results for 2 indicate that in concentrated DMF solution phen remains bonded to copper but salicylate is likely monodentate in contrast to the situation for 1. However, in dilute DMF solution, both 1 and 2 form the same species, which accounts for the similarity in anti-convulsant activity of the two compounds. Both 1 and 2 were found to be effective in preventing MES-induced seizures and ineffective in preventing Metrazol-induced seizures. Rotorod toxicity, consistent with central nervous system depression, paralleled the observed anti-convulsant activity. It is suggested that the observed anti-convulsant activity is consistent with central nervous system depression as a physiological mechanism in overcoming MES induced seizures due to MES-induced brain inflammatory disease. PMID- 11931960 TI - Nuclease activity of [Cu(sulfathiazolato)(2)(benzimidazole)(2)]2MeOH. Synthesis, properties and crystal structure. AB - The [Cu(sulfathiazolato)(2)(benzimidazole)(2)]2MeOH complex has been synthesised and characterised. It crystallises in the monoclinic system, space group C1c1, with unit cell dimensions a=18.829(7) A, b=12.206(3) A, c=17.233(5) A, alpha=90.06(2) degrees, beta=97.28(3) degrees, gamma=90.21(3) degrees and Z=4. The geometry around the copper(II) ion is intermediate between tetrahedral and square planar. The complex produces cleavage of plasmid pUC18 in presence of reducing agents. The efficiency of cleavage reaction of the title compound with pUC18 and with different reducing agents follows the order ascorbate H(2)O(2)>ascorbate>MPA>dithiothreitol>H(2)O(2). PMID- 11931962 TI - Metal binding by melanins: studies of colloidal dihydroxyindole-melanin, and its complexation by Cu(II) and Zn(II) ions. AB - Melanins are colloidal pigments known to have a high affinity for metal ions. In this work, the nature of the metal-binding sites are determined and the binding affinities are quantified. Initial potentiometric titrations have been performed on synthetic dihydroxyindole (DHI) melanin solutions to determine the chemical speciation of quinole/quinone subunits. Two types of acidic functionalities are assignable: catechol groups, with pK(a) between 9 and 13, and quinone imines (QI), with pK(a) of 6.3. The presence of the quinone-imine tautomer has, to our knowledge, never been assessed in polymeric melanins. Melanin solutions obtained from N-methylated DHI lack the pK(a) 6.3 buffer, consistent with its inability to form the quinone-imine tautomer. EPR spectroscopy of the DHI-melanin samples demonstrates that the semiquinone radical is in too low a concentration to contribute to the bulk binding of metals. Changes in the titration curves after addition of Cu(II) and Zn(II) ions were analyzed to obtain the binding constants and stoichiometry of the metal-melanin complexes, using the BEST7 program. UV-Vis spectra at neutral and high pH are used to identify absorbances due to Cu-bound quinone imine and catechol groups. The derived binding constants were used to determine speciation of the Cu(II) and Zn(II) ions coordinated to the quinone imine and catechol groups at various pH. The mixed complexes, Zn(QI)(Cat)(-) and Cu(QI)(Cat)(-) are shown to dominate at physiological pH. PMID- 11931961 TI - Synthesis, characterization and biological activity of two new polymeric copper(II) complexes with alpha-ketoglutaric acid thiosemicarbazone. AB - In this paper we describe the synthesis of new copper complexes with alpha ketoglutaric acid thiosemicarbazone. The crystal structures of the two compounds: [Cu(H(2)ct)Cl](n) [(Cu(H(2)ct)Cl)(2)] (1) and [Cu(Hct)](n).3nH(2)O (2) (H(3)ct=alpha-ketoglutaric acid thiosemicarbazone) have been determined by X-ray and spectroscopic methods. In 1 two independent copper atoms are present. Cu(1), in a nitrogen- and oxygen-bridged polymer, is a six-coordinated (4+2), Cu(2), five coordinated (4+1), is a chlorine-bridged dimer. In 2 the copper atom presents a penta-coordination, polymeric chains form layers and the CH(2)CH(2)COO(-) groups bridge copper atoms. In 1 a monodentate and in 2 a syn anti bidentate bridging carboxylate are present. The biological properties of 1 and 2 and also of the free ligand (H(3)ct) were tested in vitro and compared on Friend erythroleukemia cells (FLC) and on human leukemia cell lines K562 and U937. On the FLC cells the free ligand does not inhibit cell growth, but increases the DNA synthesis; complex 1 inhibits cell proliferation and increases the DNA synthesis; complex 2 inhibits cell growth, but induces a decrement of DNA synthesis and increases the reverse transcriptase activity. Regarding the human cell lines, both complexes show proliferation inhibition through an apoptosis mechanism on cell line U937, while they have no effects on the K562 line. PMID- 11931963 TI - Redox behavior of melanins: direct electrochemistry of dihydroxyindole-melanin and its Cu and Zn adducts. AB - Synthetic melanin films, formed on electrode surfaces by oxidative polymerization of 5,6-dihydroxyindole solution, were used to directly measure the chromophore's redox reactivity. Films on optically transparent indium-tin oxide (ITO) electrodes allow correlation of spectral changes with electrochemical potential. Spectroelectrochemical titrations show an initial reversible transformation that is ascribed to formation of a unique quinone-imine chromophore. The apparent E(1/2) for maximum quinone-imine formation is approximately 125 mV (vs. Ag/AgCl) but at potentials higher than 100 mV, an irreversible bleaching is evident. Correlation of the current with the monomer concentration implies that only one in six monomers is oxidized to the quinone-imine before the irreversible bleaching occurs. Films pretreated with CuCl(2) and Zn(CH(3)COO)(2) show elevated quinone-imine absorbances, even under reducing conditions, indicating a preferential stabilization of this state by coordination to the metals. PMID- 11931964 TI - E.s.r., visible and SOD studies of imidazolate bridged Cu(2)(II,II), Cu(II)Zn(II) and Cu(II)Ni(II) complexes with pentamethyldiethylenetriamine as capping ligand: a plausible model for superoxide dismutase. AB - X-band e.s.r. and electronic spectra of imidazolate bridged homobinuclear Cu-Cu complex, [(PMDT)Cu-Im-Cu(PMDT)](ClO(4))(3) and heterobinuclear Cu-Zn and Cu-Ni complexes, viz. [(PMDT)Cu-Im-Zn(PMDT)](ClO(4))(3), [(PMDT)Cu-Im-Ni(PMDT)] (ClO(4))(3), where PMDT=pentamethyldiethylenetriamine, Im=Imidazolate ion and related mononuclear complexes, [(PMDT)Cu(OH(2))](2+) and [(PMDT)Cu(ImH)](2+) have been described. Superoxide dismutase activities of these complexes have also been measured. PMID- 11931965 TI - Zinc-coordination of aspartic acid-76 in Sulfolobus ferredoxin is not required for thermal stability of the molecule. AB - The highly thermostable 7Fe-ferredoxin from Sulfolobus sp. strain 7 has tightly bound zinc at the interface between the N-terminal extra domain and the C terminal core. The zinc is tetrahedrally ligated by His-16, His-19, His-34, and Asp-76. Previous studies on truncated mutants have shown that the zinc and certain parts, i.e. not all, of the N-terminal extra stretch are responsible for the thermal stabilization of the molecule. To study the role of Asp-76, a series of mutants were constructed with Asp-76 replaced by Glu (D76E), Asn (D76N), or Ala (D76A). All the mutants, as well as wild type ferredoxin, bound 1 mol zinc/mol protein, and showed similar kinetics for 2-oxoacid:ferredoxin oxidoreductase. The stability of the protein was examined by thermal degradation of the clusters. In the absence of guanidium thiocyanate, the T(m), defined as the mid-point temperature of the thermal transition from the native to the denatured state, for every mutant was above 100 degrees C. The T(m) values in the presence of 1 M guanidium thiocyanate were determined to be 90.8, 90.2, 87.1, 84.4, and 72.9 degrees C for the natural, recombinant, D76N-, D76A-, and D76E ferredoxins, respectively. These results indicate that the interaction between zinc and the carboxyl oxygen of Asp-76 has subtle effects on both the zinc ligation and stability, although the native zinc center is liganded with high symmetry, suggesting that the three His residues are more important for zinc binding. PMID- 11931966 TI - Thiourea derivatives and their nickel(II) and platinum(II) complexes: antifungal activity. AB - We have synthesized two thiourea derivatives of methyl anthranilate (1, 2) and their complexes with nickel (3) and platinum(II) (4). We have also prepared the complexes of nickel(II) with two benzoylthiourea derivatives (5, 6). The obtained compounds were characterized by elemental analysis, spectroscopic methods (FT-IR, UV-vis, NMR), mass spectrometry and thermal analysis. Compound 1, C(20)H(23)N(3)O(2)S, crystallizes in monoclinic space group P21/n, with Z=4, and unit cell parameters, a=8.8042(4) A, b=7.6608(3) A, c=28.834(2) A, alpha=gamma=90 degrees, beta=90.94(1) degrees. Compound 2, C(20)H(21)N(3)O(3)S, crystallizes in monoclinic space group P21/c, with Z=4, and unit cell parameters, a=7.7345(4) A, b=8.6715(4) A, c=29.113(2) A, alpha=gamma=90 degrees, beta=90.67(1) degrees. Compound 5, C(24)H(30)N(4)NiO(2)S(2), crystallizes in monoclinic space group P21/n, with Z=4, and unit cell parameters, a=10.4317(8) A, b=18.517(2) A, c=13.299(1) A, alpha=gamma=90 degrees, beta=104.53(1) degrees. Compound 6, C(25)H(28)Cl(2)N(4)NiO(4)S(2), crystallizes with a molecule of CH(2)Cl(2) in triclinic space group P-1, with Z=2, and unit cell parameters, a=10.362(1) A, b=11.849(2) A, c=12.536(2) A, alpha=90.04(2) degrees, beta=84.73(1) degrees, gamma=113.43(2) degrees. Compounds 1 and 2 show antifungal activity against the major pathogens responsible for important plant diseases (Botrytis cinerea, Colletotrichum fragariae, Fusarium oxysporum and Phoma betae). The antifungal activity is practically the same for morpholine and ethyl derivatives. PMID- 11931968 TI - 15-MC-5 manganese metallacrowns hosting herbicide complexes. Structure and bioactivity. AB - Interaction of manganese with salicylhydroxamic ligands leads to the formation of a series of 15-membered metallacrown Mn(II)(L)(2)[15-MC(Mn(III)N(shi))-5](py)(6) (L=alkanoato ligand). The crystal structure contains a neutral 15-membered metallacrown ring of the type [15-MC(Mn(III)N(shi))-5]. The metallacrown core consists of five Mn(III) and five shi(-3) ligands. The 15-membered metallacrown ring is formed by the succession of five structural moieties of the type [Mn(III) N-O]. The diversity in the configuration (planar or propeller) for the ring Mn(III) ions gives to the metallacrown core flexibility and simultaneously allows the encapsulation of the sixth Mn(II). The encapsulated Mn(II) is seven coordinate and is bound to the five hydroximate oxygen donors provided by the metallacrown core, and two oxygen atoms from the carboxylate herbicide ligands. Antibacterial screening data showed that among all the compounds tested, manganese metallacrowns are more active than the simple manganese herbicide or carboxylate complexes while an increase in the efficiency of [15 MC(Mn(III)N(shi))-5] towards the analogous [12-MC(Mn(III)N(shi))-4] can be observed. PMID- 11931967 TI - Synthesis and selective tumor targeting properties of water soluble porphyrin Pt(II) conjugates. AB - We have designed and synthesized a series of novel water soluble porphyrins and their platinum(II) conjugates, cis-[(Por)Pt(dmso)X], where Por=5-(4-pyridyl) 10,15,20-tris(4-sulfonatophenyl)porphyrin) (PyTPPS) or 5-[4-(3 aminopropyl)pyridiniumyl]-10,15,20-tris(4-sulphonatophenyl)porphyrin (PyTPPS NPn), X=2Cl, 1,1-cyclobutanedicarboxylic acid, oxalate, or malonate. Their biodistribution in tumor bearing mouse was examined along with their antitumor activity against murine leukemia L1210 cell line. The representative complex 1 exhibited a significant accumulation in tumor tissue with a tumor/muscle ratio of 7 after 24 h post injection. The antitumor activity of the title compounds was marginal (T/C: 95-117%), but it was found that platinum(II) coordination to the porphyrin periphery did not affect the tumor accumulating properties of the porphyrin permitting further derivatization for efficient delivery of the Pt(II) antitumor agent. PMID- 11931969 TI - Spectroscopy, cytotoxicity and DNA-binding of the lanthanum(III) complex of an L valine derivative of 1,10-phenanthroline. AB - The interaction of the lanthanum(III) La(III)-L (L=N,N'-bis-(1-carboxy-2 methylpropyl)-1,10-phenanthroline-2,9-dimethanamine) complex with calf thymus DNA was studied by electronic spectra, fluorescence spectra and circular dichroic spectra. The La(III)-L complex was assayed for antitumor activity in vitro against the HL-60 (the human leucocytoma) cells, HCT-8 (the human coloadenocarcinoma) cells, BGC-823 (the human carcinoma of stomach) cells, Bel 7402 (the human liver carcinoma) cells and KB (the human nasopharyngeal carcinoma) cells. The results show that the La(III)-L complex has activity against HL-60 cells, Bel-7402 cells and KB cells. Moreover, it is slightly more effective against Bel-7402 cell line than cisplatin. Using ethidium bromide as a fluorescence probe, the binding mode of the La(III)-L complex to calf-thymus DNA was studied spectroscopically. For comparison, the same measurements were carried out with La(III)-Phen [La(III)-1,10-phenanthroline complex] and La(III)-Val [La(III)-L-valine complex]. The results indicate that the La(III)-L and La(III) Phen complexes possibly interact with calf-thymus DNA by both intercalative and coordination binding, whereas the La(III)-Val complex interacts with calf-thymus DNA by coordination binding. Kinetics of binding of the three complexes to DNA is for the first time studied using ethidium bromide as a fluorescence probe with stopped-flow spectrophotometer under pseudo-first-order condition. The strong two step mechanisms in the process of the La(III)-L and La(III)-Phen complexes and one step in the process of the complex La(III)-Val interacting with DNA are observed, and the k(obs) (observed pseudo-first-order rate constant) and E(a) (observed energy of activation) values of binding to DNA are obtained. PMID- 11931970 TI - Studies of cadmium binding to hexokinase: structural and functional implications. AB - The interaction between cadmium and yeast hexokinase was studied. Cadmium produces changes in the aggregation state of the protein and large structures with a large molecular mass were formed. This phenomenon occurs without large modifications to the secondary structure. During this change the enzyme maintains a high level of activity in the monomer as well as in aggregate form. This implies that the enzyme function is not greatly affected by the change and it maintains its active sites without significant modifications. According to kinetic measurements with both glucose and ATP as a variable substrate, cadmium causes a mixed-type inhibition with a main uncompetitive component. Binding experiments show that the protein presents negative cooperative binding with cadmium at various temperatures (298, 303 and 313 K) and a progressive loss in metal union with concentration depending on the temperature. The total union percentage decreases as the metal concentration increases. This is probably due to the aggregation process, which affects the binding sites for the metal and also for the substrate. Labile interactions are more persistent than specific interactions in accordance with the solvation parameter. PMID- 11931971 TI - Study of the influence of metal ions on tRNA(Phe) thermal unfolding equilibria by UV spectroscopy and multivariate curve resolution. AB - The influence of metal ions (Na(+), Mg(2+) and Cd(2+)) on the thermal unfolding of phenylalanine transfer ribonucleic acid (tRNA(Phe)) was studied by UV spectroscopy-monitored melting experiments. Absorbance data were obtained during the unfolding process in the range 220-340 nm and later analyzed by a multivariate curve resolution approach (MCR-ALS) based on factor analysis. This procedure determines the number of spectroscopically distinct conformations present during the unfolding process and reveals their concentration profiles and pure spectra, without any initial assumption having to be made about the number of steps in the unfolding pathway. From the concentration profiles and pure spectra, information such as T(m) values can be recovered. The results were compared with those obtained previously in spectroscopic and calorimetric unfolding experiments, showing that the multivariate approach recovers information that complements that obtained in traditional spectroscopic melting experiments. PMID- 11931972 TI - Comparative studies on the iron chelators O-TRENSOX and TRENCAMS: selectivity of the complexation towards other biologically relevant metal ions and Al(3+). AB - Complexation constants have been determined by potentiometric titration and spectrophotometric measurements for several biologically relevant divalent metals (Ca(2+), Cu(2+), Zn(2+)) as well as Al(3+) with the sulfonated tris(8 hydroxyquinolinate) tripodal ligand O-TRENSOX. The values demonstrate great selectivity of O-TRENSOX for Fe(3+) according to the sequence Fe(3+) >>Cu(2+)>Zn(2+)>Ca(2+). This selectivity is compared to that shown by tris(hydroxamate) and tris(catecholate) ligands. (1)H NMR spectroscopy of the diamagnetic complexes have been carried out in (2)H(2)O solutions. PMID- 11931973 TI - Interactions of the anticancer antibiotic altromycin B with copper(II), palladium(II) and platinum(II) ions and in vitro activity of the formed complexes. AB - Interaction of the anticancer antibiotic altromycin B with Cu(II), Pd(II) and Pt(II) ions was studied using 1H-NMR, EPR, electronic absorption and circular dichroism spectroscopy. The results derived from NMR studies where that the Pt(II) and Pd(II) ions interact with the nitrogen atom of the dimethylamino group of the C(10)-disaccharide, while the C(2)-epoxide group does not participate and remains intact. Cu(II) ions interact in a different way with altromycin B as was concluded by EPR and circular dichroism spectra. Altromycin B coordinates to the Cu(II) ions via the oxygen atoms of the C(11) phenolic and the C(12) carbonyl group while the nitrogen atom does not participate in the complexation. The presence of these metal ions improves the stability of altromycin B in solution. These complexes were studied in vitro against K562 leukemia sensitive and doxorubicin-resistant cells and GLC4 lung tumor cells, sensitive and doxorubicin resistant. The activity of the complexes compared to the free drug is improved against resistant cells and is affected moderately against sensitive cells. Finally, 20% of platinum added as altromycin B metal complex entered GLC4 cells. PMID- 11931974 TI - Vanadate-induced cell growth arrest is p53-dependent through activation of p21 in C141 cells. AB - Vanadium is widely used in industry. It is a potent toxic agent and carcinogen. The mechanisms involved in its toxicity and carcinogenesis are still unclear. Improper cell growth is believed to be involved in cancer development. The present study investigated the regulation of p53 on vanadate-induced cell growth arrest using both p53 wild type C141 cells and p53 deficient embryo fibroblasts (p53 -/-). On vanadate stimulation, C141 cells exhibited a dose- and time dependent S phase arrest as determined by DNA content analysis. In contrast, vanadate was unable to increase the percentage of S phase in p53 -/- cells. Luciferase assay showed that vanadate induced p53 activation in a dose- and time dependent manner in p53 wild type C141 cells. Addition of pifithrin-alpha (PFT), a specific inhibitor of p53, reduced the activation of p53 with a concomitant decrease in growth arrest at S phase. Western blotting analysis demonstrated that vanadate caused a dose- and time-dependent increase of p21 level in C141 cells. Pretreatment of C141 cells with PFT decreased p21 expression induced by vanadate while the p21 expression did not vary in vanadate stimulated p53 -/- cells. The results obtained from the present study suggest that vanadate is able to induce S phase arrest through p53- and p21-dependent pathway. PMID- 11931975 TI - Determination of element distribution between the symplasm and apoplasm of cucumber plant parts by total reflection X-ray fluorescence spectrometry. AB - The distribution of Cd, Ni, Pb and Fe between the symplasm (cytoplasm) and apoplasm (cell wall) of cucumber roots and leaves was determined by total reflection X-ray fluorescence spectrometry following a special sample preparation procedure. The plants were grown in modified Hoagland nutrient solution containing Fe in chemical form of Fe(III)-citrate or Fe(III)-EDTA, as well as the heavy metal contaminants, each in concentration of 10 microM. In the roots the larger part of Pb was found in the apoplasm, while Ni and Cd were mostly in the symplasm. In the leaves however, about 50-60% of the Pb content and practically the total amount of Cd were detected in the symplasm. About 30-40% of the translocated Ni remained in the apoplasm of the leaves. The Cd-, Ni- and Pb treatments resulted in higher total concentration of Fe in the roots, however, the relative amount of Fe in the symplasm decreased in all cases. In the leaves of the control plants the larger part (60-80%) of Fe occurs in the symplasm. Due to the heavy metal effects, the relative amount of Fe in the symplasm decreased except in the Pb-contaminated plants, where in the presence of Fe(III)-EDTA, the Pb treatment resulted in a moderate increment of Fe concentration in the symplasm. PMID- 11931976 TI - Comparative modeling of the latent form of a plant catechol oxidase using a molluskan hemocyanin structure. AB - The structure of the precursor form of catechol oxidase from sweet potatoes (Ipomoea batatas) has been modeled on the basis of the 3D structural data of mature catechol oxidase [Nat. Struct. Biol. 5 (1998) 1084] and of hemocyanin from giant octopus (Octopus dofleini) [J. Mol. Biol. 278 (1998) 855]. A C-terminal extension peptide is found in the cDNA sequence but not in the purified, mature form of catechol oxidase. Superimposition of the 3D structures of the native hemocyanin and catechol oxidase reveals a close relationship except for an additional C-terminal domain only found in the hemocyanin structure. As sequence alignment shows good homology this domain of the hemocyanin structure was used as a template to model the 3D structure of the C-terminal extension peptide of catechol oxidase. As hemocyanins show no or only weak catecholase activity due to this domain this indicates an inhibitory function of this extension peptide. Beside this possible shielding function for the precursor form, evidence for a function in copper-uptake also increases due to the location of three histidine residues in the model. PMID- 11931977 TI - Methylmercury induces the opening of the permeability transition pore in rat liver mitochondria. AB - Interactions of methylmercury (CH(3)HgCl) with non-energized mitochondria from rat liver (non-respiring mitochondria) have been investigated in this paper. It has been shown that CH(3)HgCl induces swelling in mitochondria suspended in a sucrose medium. Swelling has also been induced by detergent compounds and by phenylarsine, a chemical compound which induces opening of the permeant transition pore (MTP). Opening of the MTP is inhibited by means of cyclosporine A. Results indicate that the swelling induced by CH(3)HgCl, as in the case of phenylarsine, is inhibited by cyclosporine A and Mg(2+), while swelling induced by detergent compounds is not cyclosporine sensitive. This comparison suggests that CH(3)HgCl induces opening of a permeability transition pore (MTP). Since the opening of an MTP induces cell death, this interaction with MTP could be one of the causes of toxicity of CH(3)HgCl. PMID- 11931978 TI - Being partial to psychostimulant addiction therapy. PMID- 11931979 TI - Role of PAR2 in pain and inflammation. PMID- 11931980 TI - Of mice and men: advances in endothelin research and first antagonist gains FDA approval. PMID- 11931981 TI - The role of DNA adduct structure and conformation in cisplatin resistance. PMID- 11931982 TI - Build your own (virtual) cell. PMID- 11931985 TI - Potentiation of the immune system by cationic amphiphilic drugs. PMID- 11931986 TI - Problem-based learning: panacea or cult? PMID- 11931987 TI - Problem-based learning: a new pathway to competence? PMID- 11931988 TI - Problem-based learning: properly balanced learning? PMID- 11931989 TI - Problem-based learning: the Maastricht experience. PMID- 11931990 TI - NAADP: an atypical Ca2+-release messenger? AB - Recently, nicotinic acid adenine dinucleotide phosphate (NAADP) has been shown, using different techniques, to mobilize intracellular Ca2+ stores in invertebrate, lower vertebrate, plant and mammalian cells. This endogenous molecule might play an atypical role in Ca2+ signalling by coordinating the responses of other Ca2+-releasing messengers. Furthermore, radioligand binding experiments have provided an insight into how desensitization of the NAADP receptor might occur. PMID- 11931991 TI - Comparison of problem-and lecture-based pharmacology teaching. AB - Problem-based learning (PBL) is gaining interest in many medical schools. Although various approaches have been labelled PBL, it remains unclear which approach is most appropriate for pharmacology courses. Moreover, some teachers remain sceptical about whether PBL is adequate to convey the numerous facts medical students need to memorize about drugs. However, comparisons of PBL methods with conventional lecture-based learning (LBL) methods within general pharmacology courses for medical students show that PBL students are at least as successful in standardized tests but enjoy their course to a greater extent than do LBL students. PMID- 11931992 TI - Modelling the consequences of receptor-G-protein promiscuity. AB - Many G-protein-coupled receptors interact with more than one type of G protein, giving rise to extreme variability in the effects of receptor activation, depending on, for example, receptor density and desensitization, efficacy of agonists, and availability of specific G proteins. This leads to errors in interpretation of data. To facilitate understanding the consequences of receptor G-protein promiscuity, we use two simplified models to simulate such consequences. Applied to the regulation of adenylyl cyclase and phosphoinositidase, the models predict seemingly paradoxical situations and explain some phenomena that, at first sight, might seem to require the induction of agonist-specific (G-protein-selective) receptor conformations. PMID- 11931993 TI - The angiotensin-converting enzyme gene family: genomics and pharmacology. AB - Modulation of the renin-angiotensin system (RAS), and particularly inhibition of angiotensin-converting enzyme (ACE), a zinc metallopeptidase, has long been a prime strategy in the treatment of hypertension. However, other angiotensin metabolites are gaining in importance as our understanding of the RAS increases. Recently, genomic approaches have identified the first human homologue of ACE, termed ACEH (or ACE2). ACEH differs in specificity and physiological roles from ACE, which opens a potential new area for discovery biology. The gene that encodes collectrin, a homologue of ACEH, is upregulated in response to renal injury. Collectrin lacks a catalytic domain, which indicates that there is more to ACE-like function than simple peptide hydrolysis. PMID- 11931994 TI - The diversity in the vanilloid (TRPV) receptor family of ion channels. AB - Following cloning of the vanilloid receptor 1 (VR1) at least four other related proteins have been identified. Together, these form a distinct subgroup of the transient receptor potential (TRP) family of ion channels. Members of the vanilloid receptor family (TRPV) are activated by a diverse range of stimuli, including heat, protons, lipids, phorbols, phosphorylation, changes in extracellular osmolarity and/or pressure, and depletion of intracellular Ca2+ stores. However, VR1 remains the only channel activated by vanilloids such as capsaicin. These channels are excellent molecular candidates to fulfil a range of sensory and/or cellular roles that are well characterized physiologically. Furthermore, as novel pharmacological targets, the vanilloid receptors have potential for the development of many future disease treatments. PMID- 11931995 TI - Pharmacological approaches to cognitive deficits and incontinence (1899-2002): progress in geriatric care. AB - In planning for the medical needs of our growing population of older adults, it has been assumed that modern pharmacology of common geriatric conditions includes strategies not considered in previous decades. However, examination of the 1899 edition of the Merck Manual demonstrates that many important contributors to geriatric disability had already started to be recognized in the 19th century. Moreover, some of the medications recommended for these conditions in 1899 were based on the same pharmacological principles as 'state-of-the-art' management options available in 2002. PMID- 11932001 TI - Effects of benzo[a]pyrene exposure on a fish population resistant to the toxic effects of dioxin-like compounds. AB - Effects of a model polycyclic aromatic hydrocarbon (PAH) were compared in populations of the estuarine fish Fundulus heteroclitus indigenous to a reference site and one highly contaminated with polychlorinated biphenyls (PCBs) and other compounds. The fish population resident to the PCB-contaminated site is genetically resistant to those PCB congeners categorized as dioxin-like compounds (DLCs) that act through the aryl hydrocarbon receptor (AHR). In response to DLC exposures, these DLC-resistant fish showed poor inducibility for enzymes known to be regulated by the AHR pathway and important for the metabolism of xenobiotics including some PAHs that also act as AHR agonists. Therefore, a laboratory study using the model PAH, benzo[a]pyrene (BaP), was conducted to evaluate how PAHs might affect these wild fish populations that differed in their inherent sensitivities to DLCs and in their tissue concentrations of contaminants. Following BaP treatment, the activities of two xenobiotic metabolizing enzymes and the concentrations of BaP-DNA adducts, as measured using the 32P-postlabeling method, were lower in the livers of DLC-resistant than reference fish. These results suggest that DLC-resistance could provide protection following chronic exposures to PAHs from the long-term consequences of DNA adduct formation, such as cancer. Alternatively, reduced metabolism and elimination of toxic or photo activated PAHs could have acute consequences to the health and reproduction of these DLC-resistant fish and their progeny. These fish populations provide useful models to evaluate the potential costs and benefits of genetic adaptation in wildlife populations subject to anthropogenic stress. PMID- 11932002 TI - Occurrence of echinocytosis in circulating RBC of black bullhead, Ictalurus melas (Rafinesque), following exposure to an anionic detergent at sublethal concentrations. AB - The shape of the erythrocytes can be altered by a great variety of chemical agents, such as many detergents due to their amphiphilic nature. The present study examines the effect of an anionic detergent on the shape of mature, circulating catfish red blood cells. Experimental exposure to sodium dodecyl benzene sulphonate dissolved in the water of aquaria at two sublethal concentrations (1.5 and 3 ppm), for a maximum of 15 days, induced morphological changes of normal erythrocyte shape to echinocytic form. These changes were evaluated at 5, 10 and 15 days after the start of treatment, using scanning electron microscopy. The crenated erythrocytes from animals exposed to detergent appeared either with border irregularities or undulations, without distinct spicules, or with numerous short spikes. Statistical analysis, applied to the data obtained from counting altered cells in the various experimental groups, showed no significant difference between the 1.5 ppm-treated animals at the three times and the controls, whereas a significant difference was observed between 3 ppm-treated animals compared to the controls, showing significance of action of the higher dose employed at the three times. These data suggest latent erythrocyte damage. The results are discussed in the light of the extensive bibliography concerning evaginating amphiphilic compounds and the mechanisms involved in echinocyte formation, taking into account the marked differences existing between the nucleated red blood cells of fish and those biconcave, unnucleated of mammals. PMID- 11932003 TI - Partitioning of accumulated trace metals in the talitrid amphipod crustacean Orchestia gammarellus: a cautionary tale on the use of metallothionein-like proteins as biomarkers. AB - The induction of metallothionein-like proteins (MTLP) as biomarkers of trace metal pollution has been investigated in the talitrid amphipod Orchestia gammarellus, an established biomonitor of trace metal availabilities in coastal areas. Sensitivity to metal exposure, MTLP and copper and zinc concentrations have been examined in amphipods from a metal-contaminated site (Dulas Bay, Wales) and two clean sites (Millport, Scotland and Bradwell, England). Groups of 10 amphipods from Dulas Bay and Millport were exposed separately to one of a concentration series (1, 3.16, 10, 31.6, 100 microM) of Cu or Zn for 17 days at 10 degrees C. Specimens from Bradwell were exposed under similar conditions but the experiment was extended to 25 days with intermediate samplings at 5, 10 and 17 days and additional groups were exposed to a mixture of Cu+Zn (7:10 molar ratio). In addition, specimens from Millport were exposed to 1 or 31.6 microM Cd for 17 days. Amphipods from Dulas Bay, which had been chronically exposed to metals in their natural environment had not acquired any tolerance to Cu and Zn since the lowest LC 50s were registered in these samples. Whatever the origin of the amphipods experimentally exposed to metals and whatever the dose of exposure, both Cu and Zn remained approximately equally distributed between cytosolic and insoluble fractions in the amphipods, suggesting that mechanisms of metal storage were identical over the whole range of conditions. Concentrations of MTLP were higher in O. gammarellus from Dulas Bay than in those from Millport analysed directly after collection, although laboratory exposures to dissolved Cd, Cu or Zn have failed to demonstrate differences in the induction of MTLP between amphipods from the clean or contaminated sites. A potential role for metallothionein-like proteins as biomarkers is thus unlikely although it remains plausible that turnover of these proteins does increase in response to increased metal challenges, enabling MTLP to play a role in metal detoxification. PMID- 11932004 TI - Genotoxic, cytotoxic and ontogenetic effects of tri-n-butyltin on the marine worm, Platynereis dumerilii (Polychaeta: Nereidae). AB - The genotoxic, cytotoxic and ontogenetic (embryo-larval) or developmental effects of tri-n-butyltin (TBT), were investigated in Platynereis dumerilii. Following the determination of maximum tolerated dose with regard to ontogenetic effects and mortality, early life stages of P. dumerilii were exposed to a range of TBT concentrations. Subsequently, the embryo-larvae were analysed for evidence of genotoxicity and cytotoxicity. Genotoxicity was assessed using cytogenetic endpoints that included the frequency of sister chromatid exchanges and chromosomal aberrations from metaphase spreads. Cytotoxicity was evaluated by determining the proliferative rate index of the growing embryo-larval cells using 5-bromodeoxyuridine labelling of the chromosomes or fluorescence plus Giemsa staining technique. TBT-exposed embryo-larvae of P. dumerilii exhibited sensitivity similar to that of other invertebrates, indicating that P. dumerilii is a suitable ecotoxicity test species. The results also suggested dose-dependent effects for genotoxic and cytotoxic end points in relation to TBT exposure. The present study highlights the need to elucidate the relative importance of direct genotoxic and indirect effects through production of genotoxic hormonal derivatives. PMID- 11932005 TI - Acute exposure of Siberian sturgeon (Acipenser baeri, Brandt) yearlings to nitrite: median-lethal concentration (LC(50)) determination, haematological changes and nitrite accumulation in selected tissues. AB - Exposure of Siberian sturgeon (Acipenser baeri) yearlings (172.0+/-18 g; mean+/ S.D.) to several NO(2)(-)-N concentrations (0, 25, 130, 180 and 275 mg/l) was studied for 72 h in static tests. At 72 h, the median-lethal concentration of NO(2)(-)-N was 130 mg/l in water with high chloride content (130.5 mg/l). Nitrite exposure produced high levels of methaemoglobin (MetHb) but did not seem to cause mortality, as surviving fish showed higher levels (82.7+/-5.6%) than torpid specimens (60.8+/-4.5%). Levels of MetHb were unrelated to environmental and plasmatic nitrite concentrations, as both torpid and surviving fish exposed to the highest nitrite levels (275 mg/l of NO(2)(-)-N) presented similar concentrations of MetHb to those exposed to 25 mg/l of NO(2)(-)-N, thus indicating the ability of Siberian sturgeon yearlings to regulate plasmatic nitrite levels and maintain them lower than the environmental concentration of the toxicant. Nitrite exposure caused changes in the plasmatic electrolyte balance, which is characterised by extracellular hyperkalemia, high plasmatic chloride levels and low plasmatic sodium concentration. Differences between the nitrite concentration in the liver of torpid (46.3+/-9.0 mg/l) and surviving specimens (19.1+/-13.1 mg/l) exposed to several concentrations of NO(2)(-)-N suggest a significant contribution of the liver in nitrite detoxification pathways, and would thus explain a possible nitrite tolerance of Siberian sturgeon yearlings. PMID- 11932006 TI - Heat shock protein Hsp70 expression and DNA damage in Baikalian sponges exposed to model pollutants and wastewater from Baikalsk Pulp and Paper Plant. AB - Lake Baikal, a unique habitat for a great number of endemic species, is the largest freshwater reservoir in the world which is still largely unaffected by anthropogenic pollution, except for some shore regions with industrial activity. The expressions of a biomarker of exposure (heat shock protein Hsp70) and a biomarker of effect (DNA single-strand breaks) were measured for the first time in endemic Baikalian sponge species (Baikalospongia intermedia, Lubomirskia fusifera, and Lubomirskia abietina). Tissue cubes of B. intermedia and dissociated cells of L. fusifera and L. abietina reacted to temperature stress (10-16 degrees C above ambient temperature) with a time-dependent increase in expression of Hsp70. In B. intermedia, the effects of model pollutants (lead, copper, and zinc, and the organochlorines tetrachloroguaiacol, TCG, and pentachlorophenol, PCP) and of the wastewater from the final refinement and aeration reservoirs of the Baikalsk Pulp and Paper Plant (BPPP), located at the shore of the southern basin of Lake Baikal, on the expression of Hsp70 and the extent of DNA damage were investigated. It was found that lead and zinc but not copper cause a strong induction of Hsp70 in this sponge, while the frequency of DNA single-strand breaks increased after exposure to all these heavy metals tested. Induction of DNA single-strand breaks was also observed after exposure to TCG and PCP, but these compounds did not (consistently) enhance Hsp70 expression. Wastewater taken from the final water aeration pond of BPPP caused a concentration-dependent increase in Hsp70 expression in B. intermedia. However, there was no difference in the basal levels of Hsp70 between sponges collected in the shallow water at an unpolluted site near Baikalsk City and at a polluted site where the wastewaters of BPPP are discharged into the lake. There was also no clear difference in the wastewater concentration-dependent induction of Hsp70 expression between sponges collected at these sites, indicating no adaptation to continuous stress exposure. PMID- 11932007 TI - Near the edge of a chromosome's "black hole". AB - The inability of current technology to determine contiguous sequence for highly repetitive regions means that centromeres fall within multi-megabase gaps, analogous to black holes from which no information escapes. However, high resolution mapping of a human X-centromere border reveals a remarkable structure, including a gradient of alpha-satellite divergence up to the edge of the youngest and most homogeneous repeats. PMID- 11932008 TI - New alleles of Notch draw a blueprint for multifunctionality. AB - The Drosophila Notch protein is one of a family of receptors that mediate an essential and perhaps universal function in the assignation of cell fates during development. Activation of Notch by its ligand, Delta, leads to the cleavage of the intracellular domain of the receptor. This domain relocates to the nucleus, where it combines with the transcription factor Suppressor of Hairless to regulate gene expression. In a recent report, Ramain, Heitzler and colleagues reveal the existence of a second Notch signalling activity that is independent of Suppressor of Hairless and might be used to link Notch activity to that of other signalling pathways, most notably Wingless/Wnt signalling. This link might be a central element in many processes of cell-fate assignation during development. PMID- 11932009 TI - MicroRNAs: deviants no longer. AB - Almost ten years ago, the Ambros laboratory made the extraordinary discovery that a gene essential for development in Caenorhabditis elegans encoded a 22 nucleotide, untranslated RNA. Further genetic studies in this nematode revealed the existence of a second tiny RNA gene that turned out to be conserved in animals as diverse as flies and humans. Now, the Ambros, Bartel and Tuschl laboratories have proven that those odd RNAs were just the first examples of a large family of RNAs, termed microRNAs (miRNAs). Although untranslated RNA genes, such as transfer RNAs and ribosomal RNAs, perform essential housekeeping roles in all living organisms, growing numbers of other RNAs, some widely conserved across phyla and others limited to certain species, are being uncovered and shown to fulfill specific duties. The discovery of miRNAs establishes a new class of regulatory RNAs and highlights the existence of unexpected RNA genes that, although ancient, are not extinct. PMID- 11932010 TI - Modulating heteroplasmy. AB - Patients with mitochondrial DNA (mtDNA) disease usually harbor a mixture of mutant and wild-type mtDNA (a state termed heteroplasmy), and the clinical features of the disease depend on the percentage of mutant mtDNA (the "mutation load") in vulnerable tissues. Factors that modulate the mutation load are poorly understood, but recent work has started to unravel the mechanisms. In certain circumstances heteroplasmy might be regulated at the level of the individual mitochondrial genome. PMID- 11932011 TI - Identifying functional links between genes using conserved chromosomal proximity. AB - Conservation of proximity of a pair of genes across multiple genomes generally indicates that their functions could be linked. Here, we present a systematic evaluation using 42 complete microbial genomes from 25 phylogenetic groups to test the reliability of this observation in predicting function for genes. We find a relationship between the number of phylogenetic groups in which a gene pair is proximate and the probability that the pair belongs to a common pathway. Our method produces 1586 links between ortholog families substantiated by observed proximity in genomes representing at least three phylogenetic groups. Of the pairs annotated in the KEGG database, 80% are in the same biological pathway in KEGG. PMID- 11932012 TI - Insects on the rise. AB - The Insect Genomics Workshop was held in Arlington, Virginia, from 28 to 30 October 2001. PMID- 11932018 TI - A common element in epidermal expression? AB - Members of an epidermally expressed gene cluster on human chromosome 21 each contain a short sequence similar to an element that can drive ectoderm-specific gene expression. PMID- 11932019 TI - Alternative splicing: multiple control mechanisms and involvement in human disease. AB - Alternative splicing is an important mechanism for controlling gene expression. It allows large proteomic complexity from a limited number of genes. An interplay of cis-acting sequences and trans-acting factors modulates the splicing of regulated exons. Here, we discuss the roles of the SR and hnRNP families of proteins in this process. We also focus on the role of the transcriptional machinery in the regulation of alternative splicing, and on those alterations of alternative splicing that lead to human disease. PMID- 11932020 TI - The sterol-sensing domain: multiple families, a unique role? AB - The "sterol-sensing domain" (SSD) is conserved across phyla and is present in several membrane proteins, such as Patched (a Hedgehog receptor) and NPC-1 (the protein defective in Niemann-Pick type C1 disease). The role of the SSD is perhaps best understood from the standpoint of its involvement in cholesterol homeostasis. This article discusses how the SSD appears to function as a regulatory domain involved in linking vesicle trafficking and protein localization with such varied processes as cholesterol homeostasis, cell signalling and cytokinesis. PMID- 11932021 TI - Lessons from animal models of Huntington's disease. AB - Huntington's disease (HD) is an autosomal-dominant neurodegenerative disorder caused by a CAG trinucleotide repeat expansion in the HD gene. The expanded repeats are translated into an abnormally long polyglutamine tract close to the N terminus of the HD gene product, huntingtin. Studies in mouse models and human suggest that the mutation is associated with a deleterious gain of function. There is now a wide range of mouse models for HD, providing important insights into processes associated with disease pathogenesis. These models have been complemented by studies in Drosophila and Caenorhabditis elegans that have allowed the identification of possible modifier loci through suppressor screens. PMID- 11932022 TI - The anterior heart-forming field: voyage to the arterial pole of the heart. AB - Studies of vertebrate heart development have identified key genes and signalling molecules involved in the formation of a myocardial tube from paired heart forming fields in splanchnic mesoderm. The posterior region of the paired heart forming fields subsequently contributes myocardial precursor cells to the inflow region or venous pole of the heart. Recently, a population of myocardial precursor cells in chick and mouse embryos has been identified in pharyngeal mesoderm anterior to the early heart tube. This anterior heart-forming field gives rise to myocardium of the outflow region or arterial pole of the heart. The amniote heart is therefore derived from two myocardial precursor cell populations, which appear to be regulated by distinct genetic programmes. Discovery of the anterior heart-forming field has important implications for the interpretation of cardiac defects in mouse mutants and for the study of human congenital heart disease. PMID- 11932026 TI - Automated detection of low-dimensional EEG alpha-episodes. An example of application to psychopharmacological data. AB - This paper addresses the practical problems raised by the fact that correlation integrals computed from electroencephalographic signals do not scale. An automated procedure is described for alpha-waves, which is based on the known method of determining the correlation dimension from scaled structures that appear in diagrams of slope-curves. Automation, which is achieved using a moving window that detects the minima of slope-curves, and the density of minima makes it possible to vary the numerous parameters, and to implement tests. The potentialities of the automatic procedure are illustrated by results of psychopharmacological interest, based on the analysis of 22x10(3) alpha-signals, and the computation of 32x10(6) correlation integrals. Previous results concerning the existence of low-dimensional alpha-episodes, lasting in general not more than 5-6 s, are confirmed. Opposing effects upon the correlation dimension are described, at their pharmacological peak, for a dopaminergic agonist (apomorphine) and an atypical antipsychotic, a sigma-receptor ligand. The results also confirm that time reparametrization is a vital check in searches of low-dimensional attractor-ruled dynamics, and they suggest that subjects exhibiting 'high-alpha', i.e. whose alpha-waves are of amplitude A>30 microV, be selected for such investigations. PMID- 11932027 TI - Automatic pattern recognition in ECG time series. AB - In this paper, a technique for the automatic detection of any recurrent pattern in ECG time series is introduced. The wavelet transform is used to obtain a multiresolution representation of some example patterns for signal structure extraction. Neural Networks are trained with the wavelet transformed templates providing an efficient detector even for temporally varying patterns within the complete time series. The method is also robust against offsets and stable for signal to noise ratios larger than one. Its reliability was tested on 60 Holter ECG recordings of patients at the Department of Cardiology (University of Bonn). Due to the convincing results and its fast implementation the method can easily be used in clinical medicine. In particular, it solves the problem of automatic P wave detection in Holter ECG recordings. PMID- 11932028 TI - Measurement of polyethylene wear - a new three-dimensional methodology. AB - Studies in the field of polyethylene wear of the acetabular cup have been generally discussed using a two-dimensional assumption on the coronal plane; significant errors thus appear. The present study proposes a new, advanced methodology in order to accurately estimate the polyethylene wear. Through the usage of the distance between X-ray focus and film and only one follow-up, anteroposterior (AP) radiograph, a three-dimensional (3D) algorithm is introduced here. Here, 91 primary total hip joint replacements in 67 patients have been examined and three findings are obtained. Results show that the mean rate of 3D linear wear of the polyethylene estimated by the current method is 0.230+/-0.036 mm per year, that of the 2D linear wear is 0.148+/-0.028 mm per year. Moreover, the wear depth of the femoral head on the sagittal plane is 0.173+/-0.043 mm per year by the current method. This study also shows that the adoption of only one AP radiograph in the evaluation of the 3D penetration of femoral head is possible. Furthermore, the methodology proposed here is more convenient than others. PMID- 11932029 TI - A computer program to fit a family and community medicine set-up. AB - A local Personal Computer (PC) network program has been designed to improve training and patient care in Family and Community Medicine set-up. The software was designed to cope with the different clinical, preventive, promotive and statistical programs of the teaching set-up. This system serves multiple levels of patient priority, assigning each patient to an individual health team staff. The present software is designed to be used for IBM PC Pentium hardware with a Window's environment. This article is a summary of the main features of the program and its applications. Such a program is essential in comprehensive health care-related decision-making and in improving patient care. The authors recommend using similar programs in family practice and in primary health care centers. PMID- 11932030 TI - Software corrections of vocal disorders. AB - We discuss how vocal disorders can be post-corrected via a simple nonlinear noise reduction scheme. This work is motivated by the need of a better understanding of voice dysfunctions. This would entail a 2-fold advantage for affected patients: Physicians can perform better surgical interventions and on the other hand researchers can try to build up devices that can help to improve voice quality, i.e. in a phone conversation, avoiding any surgical treatment. As a first step, a proper signal classification is performed, through the idea of geometric signal separation in a feature space. Then through the analysis of the different regions populated by the samples coming from healthy people and from patients affected by T1A glottis cancer, one is able to understand which kind of interventions are necessary in order to correct the illness, i.e. to move the corresponding feature vector from the sick region to the healthy one. We discuss such a filter and show its performance. PMID- 11932031 TI - RESP-24: a computer program for the investigation of 24-h breathing abnormalities in heart failure patients. AB - In this paper, we describe a computer program (RESP-24) specifically devised to assess the prevalence and characteristics of breathing disorders in ambulant chronic heart failure patients during the overall 24 h period. The system works on a single channel respiratory signal (RS) recorded through a Holter-like portable device. In the pre-processing stage RESP-24 removes noise, baseline drift and motion artefacts from the RS using a non-linear filter, enhances respiratory frequency components through high-pass filtering and derives an instantaneous tidal volume (ITV) signal. The core processing is devoted to the identification and classification of the breathing pattern into periodic breathing (PB), normal breathing or non-classifiable breathing using a 60 s segmentation, and to the identification and estimation of apnea and hypopnea events. Sustained episodes of PB are detected by cross analysis of both the spectral content and time behavior of the ITV signal. User-friendly interactive facilities allow all the results of the automatic analysis procedure to be edited. The final report provides a set of standard and non-standard parameters quantifying breathing abnormalities during the 24 h period, the night-time and the day-time, including the apnea/hypopnea index, the apnea index, the total time spent in apnea or in hypopnea and the prevalence of non-apneic and apneic PB. The accuracy of these measurements was appraised on a data set of 14 recordings, by comparing them with those provided by a trained analyst. The mean and standard deviation of the error of the automatic procedure were below respectively 6 and 8% of the reference value for all parameters considered and the mean total classification accuracy was 92%. In most cases, the individual error was <12%. We conclude that measurements provided automatically by the RESP-24 software are suitable for screening purposes and clinical trials, although a preventive check of signal quality should be recommended. PMID- 11932032 TI - Insulin sensitivity in type 2 diabetes: univariate and multivariate techniques to derive estimates of insulin sensitivity from the insulin modified intravenous glucose tolerance test (FSIGT). AB - The use of univariate and multivariate techniques to derive estimates of insulin sensitivity from the insulin modified FSIGT were investigated in 12 Type 2 diabetic subjects aged (mean+/-S.D.) 59+/-9.5 years and BMI 28.1+/-2.2 kg m(-2), who underwent both a FSIGT and an isoglycemic hyperinsulinemic clamp. Reproducibility of the FSIGT was tested in four patients on three separate occasions. FSIGT data were assessed by both univariate and multivariate techniques. The sensitivity index for the FSIGT ranged from 0.162 to 3.292 (mean 1.378) x 10(-4) x l min(-1) mU(-1) for the univariate approach and from 0.163 to 2.727 (mean 1.378) x 10(-4) x l min(-1) mU(-1) for the multivariate method. Mean S(Iclamp) was 44.41 x 10(-4) x l(-2) min(-1) x mU(-1) (range 22.0-77.92). The correlation of the insulin sensitivity indices between the clamp and the FSIGT was 0.51 (P=0.056) for the univariate and 0.67 (P=0.017) for the multivariate analyses. Repeated FSIGTs showed a lower variability for the multivariate than for the standard approach. PMID- 11932033 TI - SIGNAL-Sequence Information and GeNomic AnaLysis. AB - An integrated software package has been developed to provide convenient graphical and textual analysis of a variety of genomic sequence features, free of charge to the biomedical research community. This package, called sequence information and genomic analysis, is available as either a stand-alone or a web-based version to enable greater versatility in access and utilization. The package can be accessed or downloaded at the following URL: http://innovation.swmed.edu/signal.htm. PMID- 11932035 TI - Notes on the history of the prion diseases. Part II. AB - The protein-only theory of transmission of the prion diseases remains controversial. Other mechanisms such as the virus, virino, and viroid hypotheses are still under consideration. All these fit in the concept of 'slow' infections that had been proposed in 1954 by Bjorn Sigurdsson, an Icelandic pathologist. Regardless of the exact mode of infection, the presence of prions in the brain has served to unite Creutzfeldt-Jakob disease (CJD), the Gerstmann-Straussler Scheinker syndrome and fatal familial insomnia, as well as scrapie and a number of other animal diseases, into a single pathological entity, the transmissible spongiform encephalopathies. The appearance of bovine spongiform encephalopathy in the United Kingdom and its putative relationship to new variant CJD, have put a new and unpredictable light on these unusual and uncommon diseases. PMID- 11932036 TI - The diagnostic interval in amyotrophic lateral sclerosis. AB - We studied whether there are any parameters that influence the period between onset of symptoms and confirmation of diagnosis in 117 patients with ALS (65 male, 52 female). The mean age of diagnosis was 57 years for men and 59 years for women. Bulbar-onset patients were older at diagnosis than limb-onset patients both men and women. Patients with bulbar-onset appeared to be more frequent in women (33:19). Contrariwise, limb-onset patients were more frequently male (43:22). The time to confirmation was much shorter with symptoms of bulbar-onset (10.5 months in male, 9.8 months in female) than for those with limb-onset (13.7 months in male, 14.8 months in female) in male, respectively, female ALS patients. The diagnosis of ALS was established in all cases by neurologists in our study. PMID- 11932037 TI - Syrinx size and duration of symptoms predict the pace of progressive myelopathy: retrospective analysis of 103 unoperated cases with craniocervical junction malformations and syringomyelia. AB - OBJECTIVE: the clinical course of syringomyelia associated with craniocervical junction abnormalities is variable. About half of the unoperated patients have benign course. This is difficult to explain on the basis of the present pathogenetic theories. Therefore, to understand the mechanism of syrinx progression better, we studied the value of syrinx size, syrinx morphology, and the syrinx/spinal cord size ratio in predicting the rate of progression and the severity of myelopathy in these patients. METHODS: 103 adult patients with syringomyelia associated with Chiari 1 malformation and/or radiographic signs of underdeveloped posterior cranial fossa were clinically and MRI examined. Patients were subdivided according to the type of disease progression. Severity of neurological deficits, and MRI features (the extent of cerebellar tonsillar displacement, anteroposterior diameter of cavities, the spinal cord and cyst/cord ratio and the shape of cavities) were measured. RESULTS: a significant relationship was found between the mid-sagittal diameter of the syrinxes and the type of disease course; patients with short duration and rapid progression had the largest diameters of cavities, whereas patients with longer duration as well as with slow rate of progression had smaller diameters (chi(2)=28.90, P<0.05; chi(2)=29.89, P<0.01; r=-0.24, P<0.05). In addition, a correlation was found between the anteroposterior diameter of the spinal cord and cyst/cord ratio and the disease duration (r=0.52, P<0.05 and r=0.48, P<0.05, respectively). CONCLUSIONS: the initial years for the development of symptomatic syringomyelia associated with CCJ malformations are characterized by relatively rapid clinical progression accompanied with distended cavities. In the later periods of unoperated syringomyelia, some patients show delay in the progress of syrinxes accompanied with collapse of cavities, probably either due to a spontaneous formation of drainage between the syrinx and the subarachnoidal space, or due to the restoration of abnormal CSF dynamics at the CCJ level. PMID- 11932038 TI - A clinicopathological study of a patient with anti-Hu-associated paraneoplastic sensory neuronopathy with multiple cranial nerve palsies. AB - Only a few cases of paraneoplastic neurologic syndrome with multiple cranial palsies have been reported. This is the case report of a patient with small-cell lung cancer and a high titer of anti-Hu antibodies who developed a tonic left pupil and multiple cranial nerve palsies, including palsies of the left fifth through tenth nerves and both twelfth nerves, as in Garcin syndrome showing at least more than seven ipsilateral cranial nerve palsies, in the course of paraneoplastic sensory neuronopathy (PSN). Pathologic examination revealed no metastasis or direct invasion of malignancy with gliosis and perivascular inflammation throughout the brainstem, indicating paraneoplastic encephalomyelitis (PEM). The numbers of EBM11+ cells (probably reactive microglia), CD8+ cells, and CD4+ cells increased. Intracellular adhesion molecule 1 and lymphocyte function associated molecule-1 were expressed intensely on the endothelia of microvessels and were found to have infiltrated mononuclear cells around microvessels in the brainstem. Multiple cranial nerve palsies and their effects including the tonic pupil are likely due to the paraneoplastic effect of the primary systemic malignancy. PMID- 11932039 TI - Surgical management of aneurysms of the bifurcation of the internal carotid artery. AB - Internal carotid artery (ICA) bifurcation aneurysms are rare and easily bleed in younger patients, but are difficult to treat surgically, due to perforators surrounding and adherent to the aneurysm. A series of 25 patients treated by clipping under the operating microscope are analyzed and compared with previous cases. Twenty-five patients, 11 men and 14 women (mean age 51 years), were treated by the same neurosurgeon. Seventeen patients presented with subarachnoid hemorrhage (Hunt & Kosnik Grade I in three, II in five, III in two, IV in seven), five with unruptured ICA bifurcation aneurysms, and three with unruptured ICA bifurcation aneurysms but another ruptured aneurysm. There were 23 small, one large, and one giant ICA bifurcation aneurysms. The projection was superior in 12, anterior in seven, and posterior in six cases. Pterional approach was employed for all cases. Outcomes were evaluated at discharge with the Glasgow Outcome Scale. Favorable outcomes (good recovery (GR) and moderate disability (MD)) were obtained in ten of 17 patients with ruptured ICA bifurcation aneurysm. Favorable outcomes were significantly greater in Grades I and II (three in I, four in II) than in Grades III and IV (one in III, two in IV; P=0.0498). Seven of eight patients with unruptured ICA bifurcation aneurysm had favorable outcomes. Temporary clipping and projection of the aneurysm did not affect the outcome. Causative factors of unfavorable outcomes were primary brain damage in cases of small and large aneurysms and perforator damage in the case of giant aneurysm. Poor clinical grade and vasospasm are the causative factors of poor outcome in patients with ruptured ICA bifurcation aneurysm. Preservation of perforators is crucial in cases of giant aneurysm. Clipping of unruptured ICA bifurcation aneurysms is recommended since they tend to bleed at a lower age than other aneurysms. PMID- 11932040 TI - Clinical course in patients with chronic carbon disulfide polyneuropathy. AB - The natural course of clinical manifestations and electrophysiological changes were studied in six patients with carbon disulfide (CS(2)) induced polyneuropathy. All of the six patients worked in the cutting-machine department of a viscose rayon plant. The environmental monitoring was also conducted in the initial stage and followed up 3 years later. In the 3-year follow-up period, the neurological symptoms and signs persisted. The highest concentration of CS(2) in the cutting machine where these patients worked was about 100-200 ppm. Three years later, the highest concentration was decreased to between 10 and 20 ppm in the cutting machine of the new production line after the engineering control had been improved. Nerve conduction velocity (NCV) studies revealed persistent abnormality in motor and sensory NCVs. Although, a tendency to improvement was noted, it did not reach a statistical significance except for conduction velocity of sural nerve in sensory NCV. Sural nerve biopsy from one patient, 2 years after diagnosis showed degeneration of both axon and myelin and a predominant loss of large myelinated fibers. A remyelination process was also noted. We concluded that CS(2) intoxication may induce a persistent damage to the peripheral nerves even after CS(2) exposure had ceased for 3 years. PMID- 11932041 TI - Frequent association with neurosurgical conditions in adult Proteus mirabilis meningitis: report of five cases. AB - Adult Proteus (P.) mirabilis meningitis is relatively rare and has not been examined individually in the English-language literature. During a period of 15 years (January 1986-December 2000), four adult patients with P. mirabilis meningitis and one adult patient with mixed bacterial meningitis involving P. mirabilis were identified at Chang Gung Memorial Hospital, Kaohsiung. These five patients included one man and four women, aged from 19 to 74 years (mean age=55.4). P. mirabilis infection accounted for 1.7% (4/229) of cases of our culture-proven monomicrobial adult bacterial meningitis and was involved in 7.1% (1/14) of cases of our adult mixed bacterial meningitis during this period. Underlying debilitating conditions including diabetes mellitus and neurosurgical disorders were common in these five cases. Adult P. mirabilis meningitis had an acute clinical course, with fever and consciousness-disturbance occurring as most prominent clinical manifestations in all patients. Other common manifestations included hydrocephalus, seizure, septic shock and wound infection. Hematogenous spread would appear to be the most likely mechanism. Multi-antibiotic resistant strains of P. mirabilis were not found in our patients. All strains were susceptible to third-generation cephalosporins, imipenem, aztreonam and ciprofloxacin. The results of treatment for adult P. mirabilis meningitis were not satisfactory, most of the patients surviving with severe neurological deficit. PMID- 11932043 TI - Primary brain abscess with Nocardia farcinica in an immunocompetent patient. AB - In this paper, we describe a case of an immunocompetent patient with cerebral nocardiosis. The onset was with loss of strength, paresthesia and focal epilepsy of the left arm. MRI showed on T2-weighted sequences a hyperintense central area of pus surrounded by a well-defined hypointense capsule and surrounding edema; on T1-weighted sequences a hypointense necrotic cavity with ring enhancement following administration of intravenous gadolinium. The patient underwent surgical excision of the abscess but culture from the specimen was negative. After 40 days of empirical antimicrobial therapy he developed neurological deterioration with focal epilepsy. A new MRI documented an enlargement of the hypointense lesion in the right frontal-parietal region. A second craniotomy with drainage of the abscess was performed; cultures yielded Nocardia farcinica. Therapy with trimethoprim/sulfamethoxazole, amikacin and meropenem was given for 35 days, and clinical and radiological improvement was observed. Home therapy was done with oral trimethoprim/sulfamethoxazole. Currently, 5 months from the second surgery, the patient can walk with support and no new episodes of epilepsy occurred. Side effects were absent from therapy. The MRI appearance of the brain lesion has improved, with a decrease in size, surrounding edema and ring enhancement. PMID- 11932042 TI - Immunohistochemical analysis of reactive astrocytes around glioblastoma: an immunohistochemical study of postmortem glioblastoma cases. AB - To investigate the mechanisms of proteolysis within the glioma, and tissue reactions against glioblastoma, immunohistochemical detection both outside and inside of the tumor was performed using seven brains with glioblastoma that were obtained from autopsies. Immunohistochemistry was performed to detect vascular endothelial growth factor (VEGF), matrix metalloproteinases (MMP)-1,-2,-9, membrane-type matrix metalloproteinase (MT-MMP), interleukin (IL)1-beta, and IL 6. The data were translated into color graphics and the localization of these proteins was analyzed. In glial cells around the tumor, GFAP, VEGF, MMP-2, and MT MMP were strongly expressed. Moreover, IL1-beta was also expressed strongly in the glial cells at the periphery of the tumor. IL-6 was recognized outside of the tumor, but was expressed only in the swollen astrocytes and normal pyramidal cells. These data suggest that in the periphery of the tumor, tissue reconstruction processes take place with concomitant degradation of the matrix by MMP-2 and MT-MMP, as well as vascular remodeling promoted by VEGF. The fact that IL1-beta, but not IL-6, was expressed strongly in the glial cells around the tumor, may indicate that these proteins expressed outside of the tumor are not utilized for tumor growth, but may be used to guard the tumor against invasions, such as immune response. PMID- 11932044 TI - Isolated oculomotor nerve palsy: diagnostic approach using the degree of external and internal dysfunction. AB - This study was performed to evaluate the recommended neuroimaging guidelines using the degree of internal and external dysfunction to avoid unnecessary catheter angiography (CA) for screening isolated oculomotor nerve palsy (IOP). We analyzed the causes and clinical characteristics of 28 consecutive patients with IOP between January 1992 and August 1999. By investigating the association of the degree of internal and external dysfunction with the etiologies of IOP, and by applying guidelines (modified by us) to these patients, we evaluated the usefulness of those guidelines. When we strictly applied the modified guidelines to our patients, CA should have been performed in 13 out of 28 patients (46.4%). These were five patients with normal pupil and partial external dysfunction (PED) due to youth or absence of pathological vascular risk factors (four ischemic; one migrainous). In addition, there were four patients with partial internal dysfunction (PID) and PED (one ischemic; one migrainous and two neoplastic); one with PID and complete external dysfunction (CED) due to young age (ischemic); one with complete internal dysfunction (CID) and PED (aneurysmal) and two with CID and CED (aneurysmal). Ten of 13 patients (76.9%) would have unnecessary CA, but no patients with aneurysms would be missed using these revised guidelines. Modified neuroimaging guidelines, using the degree of internal and external dysfunction, seem to be useful in the management of IOP. PMID- 11932045 TI - Evidence of tolerance to baclofen in treatment of severe spasticity with intrathecal baclofen. AB - In a retrospective study, changes in baclofen dose and complication frequency were recorded in 79 patients with intrathecal baclofen administration and the effect on nightly muscle spasms was measured over a mean observation period of 34 months, during which time an increase in the daily dose of baclofen during the first 1--1-1/2 years is notified. On subsequent pump fillings the daily dose of baclofen remained stable in the group of non-multiple sclerosis patients. In contrast, the group of multiple sclerosis patients showed a steady increase in their daily dose of baclofen. We found a frequency of complications of 0.014 monthly often due to catheter problems. There was a significant decrease in numbers of nightly muscle spasms in an 8 h recording period from 77+/-20 preoperatively and 9+/-3 (P=0.02) 3 months after surgery. The steady increase in the daily dose of baclofen in order to obtain adequate reduction in spasticity and nightly muscle spasms in the first 1--1-1/2 years cannot fully be explained by caution and difficulties in achieving the correct dose, but also indicates that tolerance to baclofen occurs. Complications are often due to infection or catheter problems. PMID- 11932047 TI - Successful removal of solitary intramedullary spinal cord metastasis from colon cancer. AB - A 69-year-old patient with an intramedullary metastasis from colon carcinoma is presented. Total removal of the tumor brought him 3 months of useful life. Although radiation therapy is generally favored, one should consider microsurgical resection of discrete, solitary intramedullary metastases in patients with radioresistant primary tumors. PMID- 11932046 TI - Spontaneous resolution of an idiopathic cervical direct vertebral arteriovenous fistula after partial coil embolization in a patient presenting with myeloradiculopathy. AB - A 53-year old female presented with paresis of the left upper extremity. Magnetic resonance imaging (MRI) and magnetic resonance angiography (MRA) disclosed a single high-flow vertebral arteriovenous fistula (AVF) with vertebral artery (VA) transection. The AVF was also fed by steal flow from the contralateral VA. The left posterior inferior cerebellar artery (PICA) branched just distal to the fistula. The fistula drained into the neighboring paravertebral veins and refluxed into the intradural venous systems. The dilated drainers compressed the spinal cord. Embolization was attempted at the drainer just behind the fistula orifice using platinum coils. The fistula was still fed slightly by right VA after the embolization, but spontaneous complete obliteration was achieved after one week. The clinical symptoms and signs disappeared. Although, detachable balloon embolization is the quickest and most effective procedure to obliterate a fistula, stepwise embolization using GDC can be considered, and may avoid the normal pressure perfusion break-through phenomenon. Spontaneous obliteration of the fistula after partial embolization in our case may result from intravenous embolization just behind the fistula orifice. It may therefore be a useful approach to the embolization of an AVF to begin the embolization at the venous side of the fistula. PMID- 11932048 TI - Prolonged unilateral vasodilatation and brain edema in fulminant hepatic failure, associated with symptomatic seizure. AB - We report prolonged unilateral vasodilatation and hemispheric brain edema in a 49 year-old man with fulminant hepatic failure (FHF). The patient presented with a tonic-clonic seizure caused by a hypertensive subcortical hemorrhage in the left parietal lobe. Serial computed tomography (CT) scans showed progressive darkening of the ipsilateral hemisphere, suggesting hemispheric cerebral infarction, but the patient did not show clinical signs of deterioration. Brain magnetic resonance angiography showed dilation of the large arteries of the left hemisphere. Evaluation of cerebral blood flow 7 days postictus with single photon emission CT revealed marked ipsilateral hyperperfusion. The darkening of the hemisphere was brain edema elicited by hyperperfusion. Brain edema was reversible, disappearing 14 days postictus. Hemispheric brain edema was caused by unilateral cerebral vasodilatation and resultant hyperperfusion. Although brain edema is a major complication in FHF patients and cerebral hyperperfusion is responsible for edema formation, CT findings of these patients almost invariably show a bilateral lesion. Unilateral vasodilatation and subsequent hemispheric hyperperfusion may be due to overproduction of vasodilators, already abundant in the brains of patients with severe hepatic failure, by seizure activity. PMID- 11932054 TI - A mechanism based understanding of cancer pain. PMID- 11932055 TI - The nociceptive flexion reflex in humans -- review article. AB - The nociceptive flexion reflex (NFR) is a physiological, polysynaptic reflex allowing for painful stimuli to activate an appropriate withdrawal response. NFR is easily measurable in clinical setting, and is a reliable and objective tool for measurement of an individual's pain experience. An exhaustive review of the literature, covering multiple search engines, indicates that the NFR method is valuable in studying the impact of diverse pharmacological and non pharmacological interventions on the flexion reflex, in conditions of acute pain and in healthy volunteers. More recently, the NFR method has gained particular attention as a research tool in studies of central sensitization and persistent or chronic pain. PMID- 11932056 TI - Intractable postherpetic itch and cutaneous deafferentation after facial shingles. AB - Some patients develop chronic itch from neurological injuries, and shingles may be a common cause. Neuropathic itch can lead to self-injury from scratching desensate skin. A 39-year-old woman experienced severe postherpetic itch, but no postherpetic neuralgia, after ophthalmic zoster. Within 1 year, she had painlessly scratched through her frontal skull into her brain. Sensory testing and skin biopsies were performed on itchy and normal scalp to generate preliminary hypotheses about mechanisms of neuropathic itch. Quantitation of epidermal neurites in PGP9.5-immunolabeled skin biopsies demonstrated loss of 96% of epidermal innervation in the itchy area. Quantitative sensory testing indicated severe damage to most sensory modalities except itch. These data indicate that in this patient, severe postherpetic itch was associated with loss of peripheral sensory neurons. Possible mechanisms include electrical hyperactivity of hypo-afferented central itch-specific neurons, selective preservation of peripheral itch-fibers from neighboring unaffected dermatomes, and/or imbalance between excitation and inhibition of second-order sensory neurons. PMID- 11932057 TI - Effect of kappa opioid agonists on visceral nociception induced by uterine cervical distension in rats. AB - Although uterine distension in rats results in an escape reflex, there exists no model of uterine cervical distension (UCD), the pain stimulus during the first stage of labor. The aims of this study were to develop such a model in virgin rats and to test whether peripherally restricted kappa opioid receptor (KOR) agonists (ADL 10-0101, ADL 10-0102, ADL 10-0116) inhibit responses to UCD. Under intravenous (i.v.) pentobarbital and alpha-chloralose anesthesia, fine metal rods were inserted in both uterine cervical osses through a small midline laparotomy. UCD was performed by manual separation of the rods (25-100 g). Single-unit afferent responses in hypogastric nerve or reflex rectus abdominis electromyographic (EMG) activity were determined before and after i.v. KOR agonists. UCD resulted in a stimulus-dependent increase in single-unit afferent activity. Units could be characterized as low threshold (mean threshold 6.6+/-2.7 g), or high threshold (mean threshold 55+/-8.8 g); all were C fibers, all responded to topical bradykinin. ADL 10-0116 (10 mg/kg) reduced the afferent response to UCD. Reflex EMG response occurred over a distension force range of 25 100 g, unaffected by i.v. saline. All three KOR agonists produced a dose dependent, naloxone-reversible inhibition of the EMG response with a potency relationship of ADL 10-0102 (ED50 0.04 mg/kg)>ADL 10-0101 (ED50 0.65 mg/kg)=ADL 10-0116 (ED50 0.60 mg/kg). These data support the use of acute UCD as a noxious stimulus, inducing afferent and reflex activity. Like other visceral stimuli, UCD is sensitive to inhibition by KOR agonists. PMID- 11932058 TI - Corticosteroid injections for lateral epicondylitis: a systematic review. AB - Patients with lateral epicondylitis (tennis elbow) are frequently treated with corticosteroid injections, in order to relieve pain and diminish disability. The objective of this review was to evaluate the effectiveness of corticosteroid injections for lateral epicondylitis. Randomised controlled trials (RCTs) were identified by a highly sensitive search strategy in six databases in combination with reference tracking. Two independent reviewers selected and assessed the methodological quality of RCTs that included patients with lateral epicondylitis treated with corticosteroid injection(s), and reported at least one clinically relevant outcome measure. Standardised mean differences were computed for continuous data and relative risks (RR) for dichotomous data. A best-evidence synthesis was conducted, weighting the studies with respect to their internal validity, statistical significance, clinical relevance, and statistical power. Thirteen studies consisting of 15 comparisons were included in the review, evaluating the effects of corticosteroid injections compared to placebo injection (n=2), injection with local anaesthetic (n=5), another conservative treatment (n=5), or another corticosteroid injection (n=3). Almost all studies had poor internal validity scores. For short-term outcomes (or=6 months), no statistically significant or clinically relevant results in favour of corticosteroid injections were found. Although the available evidence shows superior short-term effects of corticosteroid injections for lateral epicondylitis, it is not possible to draw firm conclusions on the effectiveness of injections, due to the lack of high quality studies. No beneficial effects were found for intermediate or long-term follow-up. More, better designed, conducted and reported RCTs with intermediate and long-term follow-up are needed. PMID- 11932060 TI - Spinal substance P release in vivo during the induction of long-term potentiation in dorsal horn neurons. AB - Long-term potentiation (LTP) in wide dynamic range (WDR) neurons in the dorsal horn has been suggested to contribute to central sensitization and the development of chronic pain. Indirect experimental evidence indicates an involvement of substance P (SP), in this respect. The aim of the present study was to monitor the extracellular level of substance P-like immunoreactivity (SP LI) in the dorsal horn of the rat during and after induction of LTP in WDR neurons in vivo. Electrophysiological recordings of single (WDR) neurons were performed in parallel with microdialysis in the dorsal horn under urethane anaesthesia. The amount of SP-LI in the microdialysate was determined by radioimmunoassay. As previously shown, high frequency conditioning stimulation of the sciatic nerve induced an increased firing response of WDR neurons. An increased response to C-fibre stimulation, but not A-fibre stimulation, could be determined. A significant increase of the extracellular level of SP-LI in the dorsal horn was detected during, but not after, induction of LTP. These data suggest that SP may be involved in the induction of LTP by high frequency stimulation. However, the maintenance of spinal LTP following high frequency peripheral nerve stimulation does not seem to depend on an increased release of SP. PMID- 11932059 TI - Neuropathic pain after C7 spinal nerve transection in man. AB - Various animal models of neuropathic pain have been developed which involve creating a lesion in a spinal root. We describe a human correlate in which patients developed a neuropathic pain syndrome after having one spinal nerve surgically divided. In some patients with brachial plexus lesions, the C7 spinal nerve from the opposite side is divided and used as a nerve transfer to re innervate the injured brachial plexus. Of five patients that underwent this procedure, one went on to develop a transient but significant neuropathic pain problem. Extensive sensory testing in this patient 2 months after surgery revealed dysesthesia and hyperalgesia to mechanical and cooling stimuli, but not to heat stimuli in the C7 dermatome of the hand on the side of C7 section. The pain and hyperalgesia persisted during a phentolamine infusion, which produced a sympathetic blockade. Only mild parasthesia persisted at a 1 year follow up. Thus, surgical division of a single spinal nerve in humans can lead to the development of neuropathic pain. PMID- 11932061 TI - Interleukin-6 in combination with its soluble IL-6 receptor sensitises rat skin nociceptors to heat, in vivo. AB - Interleukin-6 (IL-6) contributes to increased pain and hyperalgesia in inflamed tissue. We have investigated the effects of IL-6, alone or in combination with its soluble receptor (sIL-6R), on the sensitivity of nociceptors to noxious heat, using dermal microdialysis. Plasmapheresis membranes were inserted into the abdominal skin of adult male Wistar rats (n=46) and perfused with modified Ringer solution. After three control samples (20 min each), the skin area above the membrane was heated to 48 degrees C for 20 min. The stimulation was followed by two washout samples. The calcitonin gene-related peptide (CGRP) content of the dialysate was measured with an enzyme immunoassay. Heat stimulation provoked a significant CGRP increase in the dialysate. Intradermal application of IL-6 (200 ng ml-1) did not significantly alter heat-induced CGRP release. However, a significant sensitisation of the heat-induced CGRP release was observed when sIL 6R (25 ng ml-1) was applied, either alone or in combination with IL-6. Neutralisation of endogenous IL-6 with a sheep anti-rat IL-6 serum did not alter heat-induced CGRP release, but abolished the sIL-6R-mediated sensitising effect. We show that IL-6 in combination with its soluble receptor can sensitise nociceptors to heat and provide evidence for the constitutive expression of the signalling molecule gp130, but not of the IL-6-membrane-bound (specific) receptor, in nociceptors. PMID- 11932062 TI - Mechanical hyperalgesia after an L5 ventral rhizotomy or an L5 ganglionectomy in the rat. AB - An L5 spinal nerve ligation (SNL) in the rat leads to behavioral signs of mechanical hyperalgesia. Our recent finding that an L5 dorsal root rhizotomy did not alter the mechanical hyperalgesia following an L5 SNL suggests that signals originating from the proximal stump of the injured nerve are not essential. We postulate that Wallerian degeneration of L5 nerve fibers leads to altered properties of adjacent intact nociceptive afferents. To investigate the role of degeneration in sensory versus motor fibers, five injury models were examined concurrently in a blinded fashion. An L5 ganglionectomy produced a selective lesion of sensory fibers. An L5 ventral root rhizotomy produced a selective lesion of motor fibers. The three control lesions included: (1) SNL with L5 dorsal root rhizotomy; (2) L5 dorsal root rhizotomy; and (3) exposure of the L5 roots without transection (sham). Paw withdrawal thresholds to mechanical stimuli were measured at three sites in the rat hindpaw corresponding to the L3, L4, and L5 dermatomes. Both the ganglionectomy and the ventral rhizotomy produced a significant, lasting (>or=20 d) decrease of mechanical withdrawal thresholds that was comparable to that produced by the SNL lesion. The L5 dorsal rhizotomy, by itself, produced a short lasting (100 microM) at other sites of ADO interaction (A1, A2A, A3 receptors, ADO transporter, and ADO deaminase) or other (IC50 value >10 microM) neurotransmitter and peptide receptors, ion channel proteins, neurotransmitter reuptake sites and enzymes, including cyclooxygenases-1 and -2. A-286501 showed equivalent potency to inhibit AK from several mammalian species and kinetic studies revealed that A-286501 was a reversible and competitive inhibitor with respect to ADO and non-competitive with respect to MgATP2-. A-286501 was orally effective to reduce nociception in animal models of acute (thermal), inflammatory (formalin and carrageenan), and neuropathic (L5/L6 nerve ligation and streptozotocin-induced diabetic) pain. A-286501 was particularly potent (ED50=1 micromol/kg, p.o.) to reduce carrageenan-induced inflammatory thermal hyperalgesia as compared to its analgesic actions in other pain models (acute and neuropathic) and its ability to alter hemodynamic function and motor performance. A-286501 was also effective to reduce carrageenan-induced paw edema and myeloperoxidase activity, a measure of neutrophil influx (ED50=10 micromol/kg, p.o.), in the injured paw. The anti-nociceptive effects of A-286501 in the L5/L6 nerve injury model of neuropathic pain (ED50=20 micromol/kg, p.o.) were not blocked by the opioid antagonist naloxone, but were blocked by the ADO receptor antagonist, theophylline. Following repeated administration, A-286501 showed less potential to produce tolerance as compared to morphine. Thus, A-286501 is a structurally novel AK inhibitor that effectively attenuates nociception by a non opioid, non-non-steroidal anti-inflammatory drug ADO, receptor mediated mechanism. PMID- 11932068 TI - The novel N-type calcium channel blocker, AM336, produces potent dose-dependent antinociception after intrathecal dosing in rats and inhibits substance P release in rat spinal cord slices. AB - N-type calcium channels modulate the release of key pro-nociceptive neurotransmitters such as glutamate and substance P (SP) in the central nervous system. Considerable research interest has focused on the therapeutic potential of the peptidic omega-conopeptides, GVIA and MVIIA as novel analgesic agents, due to their potent inhibition of N-type calcium channels. Recently, the novel peptidic N-type calcium channel blocker, AM336, was isolated from the venom of the cone snail, Conus catus. Thus, the aims of this study were to (i) document the antinociceptive effects of AM336 (also known as CVID) relative to MVIIA following intrathecal (i.t.) bolus dosing in rats with adjuvant-induced chronic inflammatory pain of the right hindpaw and to (ii) quantify the inhibitory effects of AM336 relative to MVIIA on K+-evoked SP release from slices of rat spinal cord. Both AM336 and MVIIA inhibited the K+-evoked release of the pro nociceptive neurotransmitter, SP, from rat spinal cord slices in a concentration dependent manner (EC50 values=21.1 and 62.9 nM, respectively), consistent with the antinociceptive actions of omega-conopeptides. Following acute i.t. dosing, AM336 evoked dose-dependent antinociception (ED50 approximately 0.110 nmol) but the doses required to produce side-effects were an order of magnitude larger than the doses required to produce antinociception. For i.t. doses of MVIIA0.07 nmol, produced a dose dependent decrease in antinociception but the incidence and severity of the side effects continued to increase for all doses of MVIIA investigated, suggesting that dose-titration with MVIIA in the clinical setting, may be difficult. PMID- 11932069 TI - A rat model of bone cancer pain. AB - This study describes the first known model of bone cancer pain in the rat. Sprague-Dawley rats receiving intra-tibial injections of syngeneic MRMT-1 rat mammary gland carcinoma cells developed behavioural signs indicative of pain, including: mechanical allodynia, difference of weight bearing between hind paws and mechanical hyperalgesia. The development of the bone tumour and structural damage to the bone was monitored by radiological analysis, quantitative measurement of mineral content and histology. Intra-tibial injections of 3 x 10(3) or 3 x 10(4) syngeneic MRMT-1 cells produced a rapidly expanding tumour within the boundaries of the tibia, causing severe remodelling of the bone. Radiographs showed extensive damage to the cortical bone and the trabeculae by day 10-14 after inoculation of 3 x 10(3) MRMT-1 cells, and by day 20, the damage was threatening the integrity of the tibial bone. While both mineral content and mineral density decreased significantly in the cancerous bone, osteoclast numbers in the peritumoural compact bone remained unchanged. However, tartarate-resistant acid phosphatase staining revealed a large number of polykariotic cells, resembling those of osteoclasts within the tumour. No tumour growth was observed after the injection of heat-killed MRMT-1 cells. Intra-tibial injections of 3 x 10(3) or 3 x 10(4) MRMT-1 cells, heat-killed cells or vehicle did not show changes in body weight and core temperature over 19-20 days. The general activity of animals after injection with live or heat-killed MRMT-1 cells was higher than that of the control group, however, the activity of the MRMT-1 treated group declined during the progress of the disease. Rats receiving intra-tibial injections of MRMT-1 cells displayed the gradual development of mechanical allodynia and mechanical hyperalgesia/reduced weight bearing on the affected limb, beginning on day 12-14 or 10-12 following injection of 3 x 10(3) or 3 x 10(4) cells, respectively. These symptoms were not observed in rats receiving heat-killed cells or vehicle. Behavioural data suggest a reasonable time window for evaluation of anti-nociceptive agents between day 14 and 20 after cancer cell inoculation in this model. Acute treatment with morphine (1-3mg/kg, subcutanously (s.c.)) produced a dose-dependent reduction in the response frequency of hind paw withdrawal to von Frey filament stimulation 17 or 19 days following intra-tibial injections of 3 x 10(3) MRMT-1 cells. A significant reduction in the difference in hind limb weight bearing was also observed. Acute treatment with celebrex (10 30 mg/kg, s.c.) did not affect mechanical allodynia or difference in weight bearing in rats 20 days following treatment with 3 x 10(3) MRMT-1 cells. Although the pathophysiology of cancer pain is largely unknown, significant enhancement of glial fibrillary acidic protein (GFAP) staining in the corresponding segments of the ipsilateral spinal cord highlights the possible involvement of astrocytes. In summary, the induction of bone cancer in the rat by the syngeneic MRMT-1 mammary tumour cell line provides a valid pre-clinical model for pain associated with bone metastases. Significant mechanical hyperalgesia and allodynia develops in association with the progression of the tumour in the bone marrow cavity, while the general condition of the animal remains satisfactory. While acute treatment with morphine has some analgesic effect on hind limb sparing the selective COX-2 inhibitor, celebrex, has no influence on the pain-related behavioural changes in this model. PMID- 11932070 TI - Neurogenic hyperalgesia versus painful hypoalgesia: two distinct mechanisms of neuropathic pain. AB - Patients with sensory disturbances of painful and non-painful character show distinct changes in touch and/or pain sensitivity. The patterns of sensory changes were compared to those of human surrogate models of neuropathic pain to assess the underlying mechanisms. We investigated 30 consecutive in-patients with dysaesthesia of various origins (peripheral, spinal, and brainstem lesions) and 15 healthy subjects. Tactile thresholds were determined with calibrated von Frey hairs (1.1mm). Thresholds and stimulus-response functions for pricking pain were determined with a series of calibrated punctate mechanical stimulators (0.2mm). Allodynia was tested by light stroking with a brush, Q-tip, and cotton wisp. Perceptual wind-up was tested by trains of punctate stimuli at 0.2 or 1Hz. Intradermal injection of capsaicin (n=7) and A-fiber conduction blockade (n=8) served as human surrogate models for neurogenic hyperalgesia and partial nociceptive deafferentation, respectively. Patients without pain (18/30) showed a continuous distribution of threshold shifts in the dysaesthetic skin area with a low to moderate increase in pain threshold (by 1.52+/-0.45 log2 units). Patients with painful dysaesthesia presented as two separate groups (six patients each): one showing lowered pain thresholds (by -1.94+/-0.46 log2 units, hyperalgesia) and the other elevated pain thresholds (by 3.02+/-0.48 log2 units, hypoalgesia). The human surrogate model of neurogenic hyperalgesia revealed nearly identical leftward shifts in stimulus-response function for pricking pain as patients with spontaneous pain and hyperalgesia (by a factor of about 5 each). The sensory changes in the human surrogate model of deafferentation were similar to patients with hypoalgesia and spontaneous pain (rightward shift of the stimulus-response function with a decrease in slope). Perceptual wind-up did not differ between symptomatic and control areas. There was no exclusive association of any parameter obtained by quantitative sensory testing with a particular disease (of either peripheral or central origin). Our findings suggest that neuropathic pain is based on two distinct mechanisms: (I) central sensitization (neurogenic hyperalgesia; in patients with minor sensory impairment) and (II) partial nociceptive deafferentation (painful hypoalgesia; in patients with major sensory deficit). This distinction as previously postulated for postherpetic neuralgia, is obviously valid also for other conditions. Our findings emphasize the significance of a mechanism-based classification of neuropathic pain. PMID- 11932071 TI - Microinjection of morphine into various amygdaloid nuclei differentially affects nociceptive responsiveness and RVM neuronal activity. AB - The goal of the present study was to identify nuclei of the amygdala in which opioid-sensitive systems can act to recruit nociceptive modulatory circuitry in the rostral ventromedial medulla (RVM) and affect nociceptive responsiveness. In lightly anesthetized rats, 10 microg of morphine was bilaterally microinjected into basolateral, cortical, medial, central, and lateral nuclei of the amygdala to determine the relative influence on the activity of identified ON, OFF and NEUTRAL cells in the RVM and on the latency of the tail flick reflex evoked by noxious radiant heat. Infusions of morphine into the basolateral nuclei resulted in a substantial, naloxone-reversible increase in tail flick latency, and significantly increased ongoing firing of OFF cells and depressed that of ON cells. The reflex-related changes in cell firing were also attenuated. Morphine infusions into the cortical nuclei resulted in a small (approximately 1 s) but significant increase in tail flick latency. As with basolateral microinjections, ongoing activity of the OFF cells was increased, and although the ongoing firing of ON cells was not significantly changed, the reflex-related burst that characterizes these neurons was reduced. Microinjections in the medial nuclei again altered ongoing activity of both ON cells and OFF cells. However, the duration of the OFF cell pause and tail flick latency were unchanged. NEUTRAL cells were not affected by morphine at any site. Morphine applied within the central, medial lateral and dorsal lateral nuclei had no effect on RVM neurons or on the tail flick. Thus, focal application of morphine within the basolateral nucleus of the amygdala produced hypoalgesia and influenced RVM ON and OFF cells in a manner similar to that seen following systemic or RVM opioid administration. Opioid action within the medial and cortical nuclei also influenced RVM cell activity, but did not prevent the reflex-related OFF cell pause, and failed to alter the tail flick substantially. These observations, plus the lack of an opioid-activated influence from the central and lateral nuclei, demonstrate fundamental differences among systems linking the different amygdalar nuclei with the RVM. One way in which the modulatory circuitry of the RVM might be engaged physiologically in behaving animals is via opioid-mediated activation of the basolateral nucleus. PMID- 11932072 TI - Vestibular tactile and pain thresholds in women with vulvar vestibulitis syndrome. AB - Vulvar vestibulitis syndrome (VVS) is a common cause of dyspareunia in pre menopausal women. Little is known about sensory function in the vulvar vestibule, despite Kinsey's assertion that it is important for sexual sensation. We examined punctate tactile and pain thresholds to modified von Frey filaments in the genital region of women with VVS and age- and contraceptive-matched pain-free controls. Women with VVS had lower tactile and pain thresholds around the vulvar vestibule and on the labium minus than controls, and these results were reliable over time. Women with VVS also had lower tactile, punctate pain, and pressure pain tolerance over the deltoid muscle on the upper arm, suggesting that generalized systemic hypersensitivity may contribute to VVS in some women. In testing tactile thresholds, 20% of trials were blank, and there was no group difference in the false positive rate, indicating that response bias cannot account for the lower thresholds. Women with VVS reported significantly more catastrophizing thoughts related to intercourse pain, but there was no difference between groups in catastrophizing for unrelated pains. Pain intensity ratings for stimuli above the pain threshold increased in a parallel fashion with log stimulus intensity in both groups, but the ratings of distress were substantially greater in the VVS group than in controls at equivalent levels of pain intensity. The data imply that VVS may reflect a specific pathological process in the vestibular region, superimposed on systemic hypersensitivity to tactile and pain stimuli. PMID- 11932073 TI - Quantitative and qualitative perceptual analysis of cold dysesthesia and hyperalgesia in fibromyalgia. AB - Somatosensory perception thresholds, perceived intensity, and quality of perceptions were assessed in 20 women with fibromyalgia syndrome (FMS) and in 20 healthy age-matched female controls. All patients and controls scaled perceived intensity and described perceived quality of randomized thermal (Thermotest) and tactile (von Frey filaments) stimulation. Perceived intensity was scaled by free number magnitude estimation and interindividual comparability was accomplished by Master Scaling. Perceived quality was assessed by choosing verbal descriptors from a list. Thenar was used as a reference for each modality tested. All patients were able to reliably scale perceived intensity at thenar, as well as in pain-affected body areas. Perception thresholds for cold pain, heat pain, cold pain tolerance and heat-pain tolerance were significantly lower in patients than controls. For cold and tactile stimulation, the master scaled perceived intensities were significantly higher in patients' pain-affected areas, whereas for warmth/heat stimulation, the intensities were significantly lower. In the qualitative perceptual analysis the most striking and significant finding was the aberration of cold-evoked perceptions in all patients: most stimuli in the range of 30-10 degrees C were reported as heat or other paresthetic or dysesthetic perceptions. The perceptual quality of warmth, and of touch, did not differ from the controls. Another aberration was observed in the nociceptive range of thermal and of tactile stimulation as significantly more frequent pain-related descriptors than in controls. This indicates a general nociceptive facilitation in addition to the lower thermal pain thresholds. The combination of cold hyperesthesia, cold dysesthesia, and multimodal hyperalgesia suggests a selective pathophysiology at a particular level of integration, possibly in the insular cortex. It is suggested that the aberrations revealed by the supraliminal sensory evaluation may be generic for FMS. Particularly, the aberrations established in all patients for perceived quality and intensity in the cold sensory channel may be an additional diagnostic criterion. PMID- 11932074 TI - Acupuncture treatment of chronic low-back pain -- a randomized, blinded, placebo controlled trial with 9-month follow-up. AB - There is some evidence for the efficacy of acupuncture in chronic low-back pain (LBP), but it remains unclear whether acupuncture is superior to placebo. In a randomized, blinded, placebo-controlled trial, we evaluated the effect of traditional acupuncture in chronic LBP. A total of 131 consecutive out-patients of the Department of Orthopaedics, University Goettingen, Germany, (age=48.1 years, 58.5% female, duration of pain: 9.6 years) with non-radiating LBP for at least 6 months and a normal neurological examination were randomized to one of three groups over 12 weeks. Each group received active physiotherapy over 12 weeks. The control group (n=46) received no further treatment, the acupuncture group (n=40) received 20 sessions of traditional acupuncture and the sham acupuncture group (n=45) 20 sessions of minimal acupuncture. Changes from baseline to the end of treatment and to 9-month follow-up were assessed in pain intensity and in pain disability, and secondary in psychological distress and in spine flexion, compared by intervention groups. Acupuncture was superior to the control condition (physiotherapy) regarding pain intensity (P=0.000), pain disability (P=0.000), and psychological distress (P=0.020) at the end of treatment. Compared to sham-acupuncture, acupuncture reduced psychological distress (P=0.040) only. At 9-month follow-up, the superiority of acupuncture compared to the control condition became less and acupuncture was not different to sham-acupuncture. We found a significant improvement by traditional acupuncture in chronic LBP compared to routine care (physiotherapy) but not compared to sham-acupuncture. The trial demonstrated a placebo effect of traditional acupuncture in chronic LBP. PMID- 11932075 TI - Adrenocortical and hemodynamic predictors of pain perception in men and women. AB - Research has demonstrated that women report more pain than men, and clinical observations suggest that attenuated adrenocortical activity is associated with high pain sensitivity. The extent to which cortisol concentrations and hemodynamics contribute to gender differences in pain sensitivity has not been investigated. Thirty-four women and 31 men performed the hand cold pressor test (CPT). Participants rated their pain every 15 s during a 90-s CPT and a 90-s post CPT recovery period and reported pain using the McGill Pain Questionnaire (MPQ). Salivary cortisol samples and cardiovascular measures were collected prior to, during, and after the CPT. Women reported greater pain than men during and after the CPT and on the MPQ (Ps<0.01). CPT disrupted the expected diurnal decline in cortisol, as shown by a significant increase in cortisol concentration post-CPT (P<0.01) in men and women. Regression analyses revealed that pre-CPT cortisol concentrations predicted lower pain reports during and after CPT in men only (P<0.01). Systolic blood pressure (BP) and stroke volume correlated negatively with pain reports only in women (Ps<0.05). Controlling for potential confounding variables did not alter these relationships. The negative association between pre CPT cortisol and pain perception in men and the association between BP and pain in women demonstrate different physiological predictors of pain perception in men and women. PMID- 11932076 TI - Vagotomy prevents morphine-induced reduction in Fos-like immunoreactivity in trigeminal spinal nucleus produced after TMJ injury in a sex-dependent manner. AB - Acute injury to the temporomandibular joint (TMJ) region activates neurons in multiple, but spatially discrete, areas of the trigeminal spinal nucleus as seen by an increase in Fos-like immunoreactive neurons (Fos-LI). Pretreatment with morphine greatly reduces Fos-LI produced in the dorsal paratrigeminal area (dPa5), ventrolateral pole of the subnucleus interpolaris/caudalis (Vi/Vc-vl) transition region, and laminae I-II at the subnucleus caudalis/upper cervical cord junction (Vc/C2) suggesting a role for these areas in processing pain signals from the TMJ region. To determine if vagal afferents contribute to neural activation after TMJ injury or reduction of activity after morphine, Fos-LI was quantified in the lower brainstem and upper cervical spinal cord of intact and vagotomized male and female rats under barbiturate anesthesia. Bilateral cervical vagotomy (VgX) did not affect Fos-LI produced by TMJ injury in males or females in the absence of morphine. By contrast, morphine-induced reduction in Fos-LI produced at the Vi/Vc-vl transition region was prevented by prior VgX in males and diestrus females, but not in proestrus females. Morphine inhibition of Fos-LI produced in laminae I-II at the Vc/C2 junction region was diminished in vagotomized males compared to intact animals, but not affected in females. In an autonomic control area, the caudal ventrolateral medulla (CVLM), VgX reversed the morphine-induced reduction in Fos-LI in males and females similarly compared to their respective intact controls. These results were consistent with the hypothesis that the Vi/Vc-vl transition region plays a unique role in deep craniofacial pain processing and may integrate autonomic and opioid-related modulatory signals in a manner dependent on sex hormone status. PMID- 11932077 TI - Comment on Raphael, K.G., Widom, C.S., Lange, G., Childhood victimization and pain in adulthood: a prospective investigation, PAIN 92 (2001) 283-293. PMID- 11932079 TI - Comment on 'A cervical anterior spinal artery syndrome after diagnostic blockade of the right C6-nerve root', PJAM Brouwers et al., PAIN 91 (2001) 397-399. PMID- 11932081 TI - Spiritual healing as a therapy of chronic pain: a randomized, clinical trial. PMID- 11932083 TI - The effect of real time 2-D-echocardiography on medical decision-making in the emergency department. AB - 2-D Echocardiography (ECHO) represents an important tool for the evaluation of the Emergency Department (ED) patient with suspected cardiovascular (CV) pathology. The present study assesses the degree of effect of real time ECHO on Emergency Physician diagnosis, treatment, and disposition of CV patients and their level of confidence in these decisions. One hundred ED patients with suspected CV pathology were enrolled into this prospective, interventional study. Senior level physicians were asked their level of confidence regarding patient diagnosis, treatment, and disposition decisions before and after the ECHO was done and interpreted by a certified sonographer in the ED. Physicians were then asked if ECHO changed any of these decisions. Patient age was 56.4 +/- 15.8 (range 27-93) years. Chest pain (n = 45) and shortness of breath (n = 38) were the most common presenting symptoms. Eighty-six of the patients were admitted. There was a change in diagnosis in 37 patients, a change in treatment in 25 patients, and a change in disposition in 11 patients. Physicians indicated there was a change in confidence level post-ECHO in approximately 50% of patients. A significant change was seen in both a more and a less confident direction. Physicians were 3 times more confident regarding diagnosis, 7 times more confident regarding treatment, and 3 times more confident regarding disposition decision-making. Real time ECHO appears to have a significant level of impact on physician level of confidence and medical decision-making concerning patients with suspected cardiovascular pathology in the ED. PMID- 11932084 TI - Bupropion seizure proportion among new-onset generalized seizures and drug related seizures presenting to an emergency department. AB - Bupropion is a relatively new and popular medication with seizures as its major side effect. This drug can produce seizures with an overdose. The purpose of this investigation was to determine the relative importance of this medication as the etiology of new-onset seizures relative to other drugs and new-onset seizures in general. The study design was a retrospective case series. All new onset generalized seizures were evaluated over a 4-year period in subjects 16 years of age and older. Etiologic diagnosis was determined from the neurology consultation and all patients with new-onset seizures were admitted to the hospital as per hospital policy and received a routine chemistry screening and a neuroimaging study as a minimum. The results indicate that 17 of 279 or 6.1% of the new-onset seizures were drug related. After cocaine intoxication (6/279 or 2.2%) and benzodiazepine withdrawal (5/279 or 1.8%) seizures, bupropion (4/279 or 1.4%) was the third leading cause of drug related seizures. In addition, all the bupropion related seizures occurred in patients taking what was considered to be a therapeutic dose or 450 mg/day or less. Sleep deprivation, previous history of attention deficit disorder and bulimia, and previous heavy alcohol use were associated in three of the patients taking bupropion who had seizures. We conclude that although drug related new-onset seizures are not a common cause of seizures overall, bupropion might be a more common cause of drug related new onset generalized seizures presenting to the Emergency Department than previously thought, occurring in more than one-fifth of this subgroup of cases. Possibly, greater exclusion criteria are needed than currently recommended for the use of bupropion at therapeutic doses. PMID- 11932085 TI - Comparative study of intra-articular lidocaine and intravenous meperidine/diazepam for shoulder dislocations. AB - The purpose of this study was to compare the analgesic effectiveness of intra articular lidocaine versus intravenous meperidine and diazepam during the reduction of anterior shoulder dislocations. Patients were randomized to one of the two methods before the reduction of shoulder dislocations. Patients marked a visual analog pain scale at baseline, after anesthesia just before reduction, and at the time of discharge. Interference with the procedure caused by pain or lack of muscle relaxation, perception of adequacy of analgesia by the patient, adverse effects, and time to discharge from the Emergency Department (ED) were measured. Differences of outcomes, relative risks (RR), and 95% confidence intervals (CIs) were derived. Fifty-four patients with anterior shoulder dislocations presenting from May 21, 1998 through January 21, 1999 were included in this study; 29 were randomly assigned to receive intra-articular lidocaine (IAL) and 25 to receive intravenous meperidine/diazepam (IVMD). IAL was less effective than IVMD in relieving pre-reduction pain (p = 0.045) but equally effective in overall pain relief (p = 0.98). IAL was more effective than IVMD in shortening recovery time (p = 0.025). There was an indication favoring IVMD in terms of physician perceived muscle relaxation and patient's perception of analgesia adequacy. In conclusion, although the IVMD method appears to have some clinically and statistically significant advantages, IAL possesses some favorable features that render it to be an analgesia alternative in shoulder dislocation reduction. PMID- 11932086 TI - Symptomatic bradycardia secondary to interaction between topical timolol maleate, verapamil, and flecainide: a case report. AB - We present a case report describing an interaction between topical timolol maleate, verapamil, and flecainide resulting in symptomatic bradycardia. Adverse drug interactions, including topical medications, are relevant in the differential diagnosis of many presenting complaints. PMID- 11932087 TI - Traumatic retrobulbar hemorrhage: emergent decompression by lateral canthotomy and cantholysis. AB - Traumatic retrobulbar hemorrhage may result in acute loss of vision that is reversible when recognized and treated promptly. A case of traumatic retrobulbar hemorrhage is presented. The technique of emergent orbital decompression by lateral canthotomy and cantholysis is described. The anatomy of the lateral canthus and the surgical procedure are illustrated by gross dissection. PMID- 11932088 TI - Internal carotid artery pseudoaneurysm masquerading as a peritonsillar abscess. AB - Blunt carotid arterial injuries are uncommon. Motor vehicle crashes are the most frequent cause, but this type of vascular injury can be secondary to any direct blow to the neck, intraoral trauma, or strangulation. Types of vascular injuries include dissection, pseudoaneurysm, thrombosis, rupture, and arteriovenous fistula formation. Patients with pseudoaneurysm of the internal carotid artery will usually present with neurologic complaints, ranging from the minor to complete stroke. On physical examination, neck hematoma, bruits, pulsatile neck mass, or a palpable thrill may be found. However, in 50% of cases, no external signs of neck trauma are observed. Onset of symptoms may occur within a few hours to several months after the initial injury. Angiography is considered the gold standard for diagnosis, but carotid Doppler ultrasound recently has been shown to be very sensitive in detecting these types of injuries. Treatment of pseudoaneurysm is often surgical, with endovascular stenting. PMID- 11932089 TI - Diverticular abscess presenting as prostate abscess and missed by abdominal CT scan. AB - This is a case report of sigmoid diverticular abscess presenting as prostate abscess. Helical computed tomography (CT) scan revealed the prostate abscess but failed to demonstrate the underlying diverticular abscess. Colonic diverticular abscess can have many unusual presentations, and multiple imaging modalities may be required to reveal the underlying pathology. Abdominal CT scan is not always a definitive and absolute imaging modality for abdominal pain evaluation and cannot supplant clinical judgment in the evaluation of confusing cases. PMID- 11932090 TI - Unilateral facial paralysis occurring in an infant with enteroviral otitis media and aseptic meningitis. AB - We report the case of a four month old infant presenting to the Emergency Department (ED) with irritability and facial asymmetry following a recent bout of gastroenteritis. Physical examination revealed a unilateral peripheral facial nerve paralysis. Common in older children and adults, facial nerve palsy has rarely been described in infancy. Although historically associated with a variety of inflammatory and infectious causes, the pathogenesis remains unclear. In this infant we were able to successfully identify an underlying acute enteroviral infection. Coxsackie B5 was isolated from the middle ear fluid, cerebrospinal fluid (CSF), nasopharyngeal and rectal swabs. After myringotomy drainage of the middle ear fluid and placement of pneumatic equalization tubes, there was rapid and complete resolution of facial paralysis. PMID- 11932091 TI - Emergency department preparedness for the evaluation and treatment of victims of biological or chemical terrorist attack. AB - This study evaluated the preparedness of Emergency Departments (EDs) in the greater Philadelphia area to evaluate and treat victims of a terrorist biological or chemical agent release. All hospitals with EDs in the survey target area were included. A survey instrument consisting of 38 questions was mailed to the physician director of each ED. Fifty-four of 62 directors returned usable surveys. This represented an overall response rate of 88.5%. Deficiencies in preparedness were identified involving physician training and education, antidote stocking, written policies, interagency agreements, and decontamination facilities. The overall level of preparedness for hospital EDs responding to this survey was low based on a set of predetermined, implicit criteria. Comprehensive plans should be developed and implemented to remedy the identified deficiencies. PMID- 11932092 TI - Low back pain. PMID- 11932093 TI - Crusted (Norwegian) scabies. PMID- 11932094 TI - A woman with an expanding annular rash. PMID- 11932095 TI - The utility of ultrasound in a case of femoral artery pseudoaneurysm and femoral arteriovenous fistula. PMID- 11932096 TI - Is the patient dead: CT scan diagnosis. PMID- 11932097 TI - Initial historical descriptions of the angina pectoris. AB - The first description of angina pectoris available in history was given in the 17th century by Edward Hyde (1609-1674), a nonmedical person, in his biography, Life of Edward, Earl of Clarendon. The case described is an in-detail history of his father's ailment, which eventually resulted in his sudden death. Although the word Angina Pectoris was not used in this manuscript, the description is very suggestive of it. The first detailed account of the angina pectoris given by a medical person was by Dr. William Heberden (1710-1801) in the 18th century. He used the term Angina Pectoris (Pectoris Dolor) for the first time. Both of these initial historical descriptions of angina pectoris are portrayed. PMID- 11932098 TI - Re: Ma, et al., Electrocardiographic manifestations: digitalis toxicity and Kumar, et al., A rare cause of congestive heart failure, The Journal of Emergency Medicine, 2001; 20:145-52, 153-7. PMID- 11932101 TI - Re: Henderson, Academic productivity in emergency medicine, The Journal of Emergency Medicine, July 2001. PMID- 11932103 TI - Pharmacologic misadventure resulting in hypercalcemia from vitamin D intoxication. PMID- 11932104 TI - The moon and the stones. Can the moon's attractive forces cause renal colic? PMID- 11932105 TI - The emergency medicine rotation: a unique experience for medical students. AB - The objective of this study was to determine if an Emergency Medicine (EM) rotation for medical students offers a unique educational opportunity, and to document those experiences. Thirty-three medical students at one teaching hospital recorded in a computer database information about their patient encounters during EM and Internal Medicine (IM) rotations. Data collected included the types of patients seen, the level of participation in patient care and decision making, and procedures performed. A total of 2740 patient encounters were recorded, 1564 EM and 1176 IM. Students on EM rotations were more likely than students on IM rotations to be involved in the initial evaluation (93.1% vs. 47.0%, respectively), diagnosis (93.5% vs. 44.7%, respectively), and decision making (93.3% vs. 43.5%, respectively); they were also more likely to perform procedures (31.7% vs. 8.5%, respectively). There were significant differences in the patient populations and disease processes encountered on the two rotations as well. PMID- 11932106 TI - Health care use by perpetrators of domestic violence. AB - Research has demonstrated that men will admit to the perpetration of domestic violence (DV), if questioned, in the medical setting. The opportunity to identify DV perpetrators, however, also depends on the frequency of contact between health care providers and perpetrators. The purpose of this study was to determine health care use among a group of DV perpetrators. A survey was administered to 133 men enrolled in the largest community-based batterer treatment program in a metropolitan region. Of the 133 men surveyed, 56 (42%) indicated they had visited a doctor, hospital, or received some other type of medical care within the preceding 6 months. Of men reporting health care visits, the majority (41%, n = 23) indicated that the Emergency Department was the location where care was provided. These data demonstrate that Emergency Departments are often visited by DV perpetrators in this community and may be important sites for screening and intervention protocols. PMID- 11932113 TI - Injury and violence prevention: a primer. AB - Unintentional and intentional injuries cause a great deal of human suffering throughout the world. They exact a huge toll on societies in terms of mortality, years of potential life lost, disability, and health care costs. The good news is that great strides have been made in understanding the causes of injuries and how to prevent them. Using seat belts and car seats, installing air bags in cars, replacing dangerous playground equipment, enforcing drinking and driving laws are but a few examples of modifying behavior, products, and environments to reduce injury risk. This paper provides an overview of the science of injury control and selected examples of how professionals in the field of patient education and counseling can contribute to enhancing the safety of the public. PMID- 11932114 TI - Still falling: a community-wide infant walker injury prevention initiative. AB - Despite the well-known risk of injury associated with use of infant walkers, they remain popular, leading to large numbers of walker-related injuries. A coalition of health care providers and educators, with the assistance of retailers and medical and human service agencies, undertook an intensive multifaceted, community-wide intervention to educate the general and health care public regarding the dangers of infant walker use and thereby reduce the number of walker-related injuries in our community. Following this intervention, 28% fewer children presented annually at the two area pediatric emergency departments for walker-related falls down stairs than during the 30 months before the intervention. The magnitude of this reduction attributable to the intervention, however, is uncertain, as national trends during the study period revealed a similar decrease in walker-related injuries. Educational interventions alone may significantly reduce but not eliminate walker-related injuries; national policy measures are likely also necessary. PMID- 11932116 TI - Increasing the use of bicycle helmets: lessons from behavioral science. AB - Bicycle helmet purchase, use, consistent use, and correct use are determined by a complex set of factors. Behavioral theory suggests that they are influenced by the reciprocal association between individual characteristics such as, expectations, skills, attitudes, and beliefs; social influences such as social norms and peer pressure; and environmental factors such as availability, accessibility, and cost. These factors can be influenced through counseling and other interventions. While a review of the literature suggests that many bicycle helmet programs have not been planned using behavioral models and knowledge from the behavioral sciences, many studies include information that supports behavioral principles. This paper describes the behavioral principles and their application to the problem of increasing bicycle helmet use. Recommendations for practitioners are included. PMID- 11932115 TI - Baby, Be Safe: the effect of tailored communications for pediatric injury prevention provided in a primary care setting. AB - Injuries are a major cause of morbidity and mortality to young children. The provision of individually tailored educational materials in primary care settings may be an effective and efficient way to promote adoption of injury prevention measures by parents. A randomized controlled study compared the effectiveness of tailored and generic persuasive communications delivered in a primary care setting on the adoption of home and car safety behaviors. During routine well child visits, a primarily African-American sample of parents of children ages 6 20 months (n=213) was randomized to receive either tailored or generic information regarding the prevention of injuries to their child. At follow-up, participants who received tailored information reported greater adoption of home and car safety behaviors than those receiving generic information. In addition, within the tailored information group, those who discussed the information with their physician showed significantly greater change than those who did not. However, this difference was not observed among those receiving generic information. Findings support the use of office-based tailored injury prevention education as a component of routine well-child care. PMID- 11932117 TI - Preventing sports injuries: opportunities for intervention in youth athletics. AB - Participation in youth sports has steadily grown over the past 30 years and continues to rise. During the 1998-1999 school year over 360,000 collegiate athletes and almost 6.5 million high school athletes participated in sports. This expansion has been accompanied by an increased awareness of the injury problem associated with participation in youth sports. Estimates are that one-third of high school athletes will sustain an injury during a sports season serious enough to result in time lost from participation. While there may always be some risk associated with sports participation, health professionals can actively encourage injury prevention. In this paper, we describe the benefits of sport participation, the injury problem associated with sports, injury prevention frameworks, and conclude by discussing the changing role of the team physician in youth sports. PMID- 11932118 TI - Surviving physical and sexual abuse: what helps low-income women? AB - OBJECTIVES: This paper contributes to the growing literature on help-seeking among women who experience violence. METHODS: We interviewed 390 low-income women who reported experiencing physical or sexual violence during adulthood, half of whom were infected with HIV. We obtained information on actions taken in response to episodes of violence and for leaving abusive relationships. RESULTS: A substantial proportion of women could predict an oncoming episode of violence; although the majority took some action either before or during the violent episode, almost 20% took no action at that time to protect themselves. Although family and friends were the greatest source of help for leaving violent relationships, the majority of women who left their abusers, 70%, did not receive assistance. Jobs and education were identified services that would help women leave abusive relationships. CONCLUSIONS: There is need for more services for low income women experiencing violence that focus on safety and on supporting and promoting financial independence. PMID- 11932119 TI - Increasing physician comfort level in screening and counseling patients for intimate partner violence: hands-on practice. AB - A paucity of literature exists on implementing and evaluating residency curriculum addressing intimate partner violence. We used unknown simulated patients in a university-based family practice clinic following a pilot curriculum intervention. The curriculum focused on physician comfort with screening, counseling, and referral of patients, using standard conferences as well as a role-play session. Subsequently, unknown simulated patients were inserted into residents' clinic schedules during videotaped sessions. Evaluation included skills checklists from simulated patients, review of videotapes, and post-study resident interviews. Use of unknown simulated patients encouraged residents to consider and screen for intimate partner violence. Using simulated patients is logistically complex but provides a powerful residency training tool. Residents reported attitude changes favoring a more comprehensive role and reported greater comfort and confidence with screening and counseling. PMID- 11932120 TI - The domestic violence survivor assessment: a tool for counseling women in intimate partner violence relationships. AB - Seeking to end violence and distress in their relationship is the goal of women abused by intimate partners. The theoretical framework guiding development of the domestic violence survivor assessment (DVSA) was Landenburger's theory of entrapment and recovery. Social context and need to balance care for others and herself influence women's decision-making about abuse. The DVSA was developed collaboratively between researchers and counselors to gain a deeper understanding of battered women's cognitive states in order to assist them during counseling to effectively resolve the dilemma of their abusive relationships while experiencing personal growth. Five states are identified which a woman may experience on 11 issues concurrently at the personal, relationship or social context levels. Research to validate the DVSA and suggestions on use with women desiring to preserve their relationship or preserve their self or preserve the resolution of change is described. Using the DVSA for assessment, intervention and measuring intermediate outcomes is delineated. PMID- 11932121 TI - The key to good healthcare communication. PMID- 11932122 TI - Analyzing medical dialogues: strength and weakness of Roter's interaction analysis system (RIAS). AB - Roter's interaction analysis system (RIAS) is analyzed in this article. Ground rules of linguistic interaction analysis, emphasizing meaning as a product of interaction and turn taking as a basic principle for the understanding of interaction are briefly introduced. Specific aspects of the application of RIAS are discussed and a number of adjustments and/or specifications suggested: (1) utterances should be defined in terms of content and turn taking criteria; (2) the recording system should allow for registering interruptions; (3) pauses or silences should be scored on the basis of functional criteria and not as demarcation in the communication; (4) clear distinctions should be made between the categories of "backchannel" and "agree"; (5) questions should be coded according to function rather than linguistic form; (6) some of the socioemotional categories may appear too narrow, others too wide; (7) crying should be included in the coding scheme as a separate category. PMID- 11932123 TI - The Roter interaction analysis system (RIAS): utility and flexibility for analysis of medical interactions. AB - The Roter interaction analysis system (RIAS), a method for coding medical dialogue, is widely used in the US and Europe and has been applied to medical exchanges in Asia, Africa, and Latin America. Contributing to its rapid dissemination and adoption is the system's ability to provide reasonable depth, sensitivity, and breadth while maintaining practicality, functional specificity, flexibility, reliability, and predictive validity to a variety of patient and provider outcomes. The purpose of this essay is two-fold. First, to broadly overview the RIAS and to present key capabilities and coding conventions, and secondly to address the extent to which the RIAS is consistent with, or complementary to, linguistic-based techniques of communication analysis. PMID- 11932124 TI - The future of intercultural mediation in Belgium. AB - Intercultural mediation was developed to solve problems between western (Belgian) health professionals and Turkish, Moroccan and Italian clients. The need for intercultural mediation in Belgium Health Care was measured by asking intercultural mediators to complete a questionnaire about situations in which language, culture, social-economic and personal circumstances cause difficulties. Results show that the profession 'intercultural mediator' continues to be important in improving the quality and accessibility of the Belgian health care for ethnic minorities. Even if clients speak Flemish fluently, there are still difficulties between health professional and client caused by culture, social economic and personal circumstances. PMID- 11932125 TI - Implementation of a nurse-monitored protocol in a Brazilian hospital: a pilot study with cardiac surgery patients. AB - The aims of this article are to describe the implementation of a nurse-monitored protocol and the adherence to cardiac therapeutic regimen of 17 male patients after coronary artery bypass graft (CABG) surgery. The protocol was developed to cover two recovery phases of the patients: during the hospitalization after CABG surgery and 6 months following hospital discharge. Follow-up included individual patient counseling through personal contacts and telephone calls. Adherence to therapeutic regimen (dietary counseling, walking program, body weight control, smoking cessation and pharmacological therapy) was self-reported. The results showed that patients reported high percentages of adherence to walking program (88.2%), smoking cessation (76.5%), dietary counseling (70.6%), and drug orientation (64.7%). However, at follow-up, there was no statistically significant difference between the group that reported adherence to medical regimen and the group that reported no adherence in use of lipid-lowering drugs (P=0.68), decrease of body mass index (P=0.28), and HDL-cholesterol level (P=0.07). PMID- 11932126 TI - Telephone counseling in psychosocial oncology: a report from the Cancer Information and Counseling Line. AB - Providing psychosocial counseling services to cancer patients and their significant others by telephone is emerging as an alternative to traditional (in person) counseling programs in psychooncology. In this paper, data are reported describing the clients of such a program that has been in continuous operation since 1981: the Cancer Information and Counseling Line (CICL) of the AMC Cancer Research Center. An examination of call record forms completed between 1 June 1998 and 30 May 1999 (N = 1627) revealed that the vast majority of callers were female (77%), non-Hispanic White (77%), with at least some college education (62%). Only 27% were cancer patients/survivors, compared to 43% who were spouses, other relatives and friends of cancer patients/survivors, and 16% who were symptomatic callers. Breast cancer was by far the most frequently mentioned cancer site (30%). Although initial topics of inquiry were dominated by requests for medical information (77%), with only a small percentage of callers initially requesting psychosocial support and counseling (12%), by the time, the call was completed, 67% had received some form of psychosocial support and/or counseling. Recommendations for future research are discussed within the context of this review. PMID- 11932127 TI - The interaction between physician and patient communication behaviors in Japanese cancer consultations and the influence of personal and consultation characteristics. AB - The communications of physician and patient vary with the characteristics of patient and consultation, as well as the communications of the counterpart. The purpose of this study is to explore the interaction between physician and patient communications in Japanese cancer consultation in view of the influence of patient and consultation characteristics. One hundred and forty cancer outpatients and 12 physicians were included in this study. The Roter Interaction Analysis System (RIAS) was used to analyze the physician-patient interaction. Patient information giving was positively related to physician facilitation, while patient question asking and emotional expression were associated with the warm and empathetic attitude of the physician. On the other hand, the encouraging statements of the physician were greater in shorter consultations, which implies physicians might have interrupted patients with encouragement before thoroughly listening to the patients concern. Further investigation is needed to confirm the causal relationships of these interactions. PMID- 11932128 TI - Facilitators and barriers to heart failure self-care. AB - Self-care of heart failure (HF) is difficult to master, but the reasons why remain unclear. The purpose of this study was to explore how HF influences patients' lives, assess how they perform self-care, and determine how their life situation facilitates or impedes HF self-care. Qualitative data were obtained from 26 individuals with chronic HF. Data were gathered using structured interviews and analyzed using content analysis. Physical limitations, debilitating symptoms, difficulties coping with treatment, lack of knowledge, distressed emotions, multiple comorbidities, and personal struggles were common. Self-care involved the recognition of symptoms, but atypical symptoms such as faintness were rarely attributed to HF. Patients discussed their successes and failures in following dietary, exercise, and medication recommendations. Some adaptation strategies were practical and some involved internal resources. Many patients accepted support from others, but some withdrew. With the number of barriers these patients face, it is not surprising that self-care of HF is typically poor and that readmission rates continue to be high. Recommendations are provided for a stepped approach to patient education and counseling that uses these findings in practice. PMID- 11932129 TI - Development and evaluation of an interactive CD-ROM for children with leukemia and their families. AB - To meet the need for an interactive software product to educate children with leukemia, ages 4-11 years, and their families about the disease and its treatment, we developed and evaluated an interactive, comprehensive, multimedia CD-ROM product, Kidz with Leukemia: A Space Adventure. The prototype was tested using a randomized controlled experimental design. Children with leukemia and their parents were randomized to receive either the newly developed CD-ROM or the book You and Leukemia by Lynn Baker. Health care providers (HCPs) and other content/technical experts evaluated only the CD-ROM. Data were collected on children's health locus of control, their understanding of leukemia, and the satisfaction of participants with their assigned intervention. Children in the CD ROM group, compared with those in the book group, showed increased feelings of control over their health. Although there was a high level of satisfaction with the CD-ROM among all users, younger children and their parents were most satisfied. In conclusion, the CD-ROM, Kidz with Leukemia: A Space Adventure, was found to be a useful, engaging, and empowering tool for children with leukemia and can serve as a model for developing future health-related educational materials. PMID- 11932130 TI - A nationwide survey of hepatitis C services provided by drug treatment programs. AB - Drug treatment programs are a site of opportunity for the delivery of primary and secondary hepatitis C (HCV) prevention services to drug users, a population at great risk for contracting and transmitting the virus. Using data collected from a random nationwide sample (N = 439) of drug treatment programs in the United States, this study examines the extent to which various types of HCV services are provided to their patients. Findings indicate that the majority of drug treatment programs educate at least some of their patients about HCV, and provide some type of support for patients who are infected with the virus. Only 29 of the programs in the sample test all of their patients for HCV, however, and 99 programs test none of them. For the most part, residential treatment programs offer more HCV related services than outpatient drug-free programs. PMID- 11932131 TI - Evaluating component effects of a prison-based treatment continuum. AB - A continuum of correctional-based therapeutic community (TC) treatment programs for drug-involved offenders has been functioning for several years in Delaware. Previous evaluations have shown the efficacy of the full continuum for up to three years posttreatment, though there has been some question of the benefits of treatment within prison. The particular focus here is on the relative impact of the within-prison, transitional, and aftercare treatment components upon criminal recidivism and relapse to illicit drug use. The relative benefit of participation in each component is supported, over and above the effects of differences in demographics and histories of criminal behavior and illicit substance use. However, the residential transitional program effects are generally larger and more long lasting. Additionally, the two outcomes appear differentially sensitive to the degree of completion of the continuum. PMID- 11932132 TI - A large randomized placebo controlled study of auricular acupuncture for alcohol dependence. AB - We report clinical data on the efficacy of acupuncture for alcohol dependence. 503 patients whose primary substance of abuse was alcohol participated in this randomized, single blind, placebo controlled trial. Patients were assigned to either specific acupuncture, nonspecific acupuncture, symptom based acupuncture or convention treatment alone. Alcohol use was assessed, along with depression, anxiety, functional status, and preference for therapy. This article will focus on results pertaining to alcohol use. Significant improvement was shown on nearly all measures. There were few differences associated with treatment assignment and there were no treatment differences on alcohol use measures, although 49% of subjects reported acupuncture reduced their desire for alcohol. The placebo and preference for treatment measures did not materially effect the results. Generally, acupuncture was not found to make a significant contribution over and above that achieved by conventional treatment alone in reduction of alcohol use. PMID- 11932133 TI - Clinicians' views on treating posttraumatic stress disorder and substance use disorder. AB - The dual diagnosis of posttraumatic stress disorder (PTSD) and substance use disorder (SUD) is reported to be both highly prevalent and highly challenging. In this study, 147 clinicians were surveyed on their degree of difficulty and gratification in working with each disorder (PTSD, SUD) and their combination; specific types of difficulties and gratifications; and personal and professional characteristics. The dual diagnosis was perceived as more difficult than either disorder alone; but, interestingly, gratification in the work was higher than its difficulty. Areas of greatest difficulty were clients' self-destructiveness, case management, and dependency; areas of greatest gratification were teaching new coping, developing expertise, and helping clients achieve abstinence. In general, difficulty and gratification appeared to be separate constructs, rather than simply opposites. Those finding the work most difficult were more likely to be in a mental health setting and to have no personal history of trauma. Clinical implications are discussed. PMID- 11932134 TI - Behavioral couples therapy for drug-abusing patients: effects on partner violence. AB - Using data from a previous investigation, the purpose of the present study was to examine the effect of Behavioral Couples Therapy (BCT) on the prevalence of partner violence among married or cohabiting substance-abusing men (N = 80). Participants were randomly assigned to receive either BCT or individual-based treatment (IBT). The proportion of couples who engaged in male-to-female physical aggression was not different during the year before treatment for dyads in BCT (n = 17, 43%) and IBT (n = 19, 48%). However, a smaller proportion of couples in the BCT condition reported male-to-female physical aggression during the year after treatment (n = 7, 18%) than those in the IBT condition (n = 17, 43%). Dyadic adjustment, frequency of heavy drinking, and frequency of drug use during the year after treatment mediated the relationship between type of treatment and the prevalence of male-to-female physical aggression. PMID- 11932135 TI - Suicide prevalence in chemical dependency programs: preliminary data from a national sample, and an examination of risk factors. AB - Completed suicides over a one year period of time were reported from a nationwide survey of Department of Veteran Affairs medical centers. Of a total of 248 completed suicides, 11 occurred in outpatient substance abuse programs, and an additional 5 occurred among patients receiving combined outpatient substance abuse and psychiatric treatment. There were no inpatient suicides. During this time, there were 7 suicide attempts on inpatient units and 37 suicide attempts in outpatient chemical dependency treatment. The majority of suicides were committed by males who had a primary alcohol addiction (63%). Thirty-eight percent of the sample had a comorbid mood disorder and 38% had a comorbid personality disorder. Risk factors relating to the potential for suicide in chemical dependency programs are discussed. PMID- 11932136 TI - Drugs and sexual effects: role of drug type and gender. AB - This study investigated gender differences in the relationship between psychoactive substance use and sexual thoughts, feelings, and behaviors. Participants (N = 464) were male and female alcohol, opiate, cocaine, and methamphetamine users enrolled in an outpatient treatment program at any of 8 sites. A self-report survey that inquired about the specific sexual thoughts, feelings, and behaviors of the participant during previous instances of being under the influence of their primary drug of dependence served as the data source. The results indicate that different categories of psychoactive agents were associated with different effects on sexual behavior, and that those effects vary by gender. Development of a valid measure assessing the type and strength of these relationships may be beneficial for use by treatment programs in promoting abstinence from drug and alcohol use and preventing relapse. PMID- 11932137 TI - DNA vaccines against cytomegalovirus: current progress. AB - The development of a vaccine for the prevention of primary cytomegalovirus (CMV) infection is a major public health priority. Live attenuated virus, recombinant viral vector, recombinant protein and peptide vaccines have been studied as potential vaccine candidates. In recent years, DNA vaccination strategies have been developed for many pathogens, including CMV. This review aims to bring together many aspects of this relatively new vaccine technology as applied to current research into the development of vaccines against CMV. PMID- 11932138 TI - Comparative in-vitro activity of penicillin alone and combined with gentamicin against clinical isolates of Streptococcus pneumoniae with decreased susceptibility to penicillin. AB - The worldwide emergence of Streptococcus pneumoniae with decreased susceptibility to penicillin has led to the suggestion that drug combinations might be used. The aim of this study was to determine possible synergy using a combination of penicillin with sub-inhibitory doses of gentamicin against 26 clinical isolates of S. pneumoniae with decreased susceptibility to penicillin, using half chequerboards and killing curves. Synergy was demonstrated for ten of the 26 isolates with the combination of penicillin with gentamicin at 1 mg/l and for 22 isolates with penicillin and gentamicin at 2 mg/l. Killing curves on three isolates showed synergy and confirmed the chequerboard results. Further synergy studies using penicillin or cefotaxime/ceftriaxone, plus low dose gentamicin against penicillin-resistant pneumococci are indicated. PMID- 11932139 TI - Effects of gatifloxacin on phagocytosis, intracellular killing and oxidant radical production by human polymorphonuclear neutrophils. AB - The ingestion and killing of bacteria by phagocytic cells is an important step in the sequence of interactions between invading microorganisms and host defense systems and may be affected by antibiotics. We investigated the effects of gatifloxacin on the phagocytosis, killing and oxidative bursts of human polymorphonuclear neutrophils (PMNs). The percentage phagocytosis and the phagocytosis index were unaffected by exposure of Escherichia coli strains to sub MICs of gatifloxacin to a 1/64 dilution. However a significant increase in percentage intraphagocytic killing and the killing index occurred in one E. coli strain at 1/32 MIC and in two strains at 1/16 MIC. The incubation of PMNs with sub-MICs and supra-MICs of gatifloxacin (to 32 MIC) did not affect the oxidative bursts. PMID- 11932140 TI - The safety of azithromycin in the treatment of adults with community-acquired respiratory tract infections. AB - The comparative safety of azithromycin was assessed in adult patients (> or =12 years) with community-acquired respiratory tract infections. Of 3229 patients evaluated, 1616 received azithromycin 500 mg once daily for 3 days and 1613 received standard regimens of amoxycillin, amoxycillin/clavulanic acid, cefaclor, clarithromycin, or roxithromycin. A similar incidence of treatment-related adverse events occurred with azithromycin (10.3%) and comparators (11.5%). Significantly fewer patients were withdrawn from azithromycin than comparator treatment (0.4 versus 2.1%; P=0.0001). Most adverse events were mild/moderate in intensity and affected the gastrointestinal system. Azithromycin was as well tolerated as other antibiotics commonly used for bacterial infections in adults. PMID- 11932141 TI - Nizatidine and omeprazole enhance the effect of metronidazole on Helicobacter pylori in vitro. AB - Treatment failures are common in patients infected with metronidazole-resistant Helicobacter pylori in the gastric mucosa when triple therapy including metronidazole is used. In patients with treatment failure and metronidazole resistant H. pylori, a higher eradication rate for H. pylori was found after secondary treatment with bismuth/ranitidine in combination with antibiotics including metronidazole, compared with the same antibiotics combined with a standard dose of omeprazole. This agrees with our previous finding that bismuth was able to reduce the susceptibility of H. pylori to metronidazole. In this study, we have found that nizatidine, an H(2)-receptor antagonist, is also able to reduce the susceptibility of H. pylori to metronidazole in vitro, despite having no direct inhibitory effect on the growth of H. pylori. This agrees with earlier findings that compounds having the ability to reverse antibiotic resistance do not necessarily have an antibiotic or chemotherapeutic effect in the sense of growth inhibition. Therefore, it was decided to investigate the effect of nizatidine and omeprazole on the oxidative respiratory chain, as it is known that metronidazole is able to inhibit the activity of fumarate reductase of H. pylori. This enzyme is a key enzyme in the alternative respiratory chain under anaerobic conditions. Nizatidine was, in these preliminary experiments, found to inhibit fumarate reductase in a dose-dependent way, like metronidazole, whereas omeprazole had almost no effect on fumarate reductase. No other significant effects on the enzymes of the respiratory chain were found. The synergistic effect of nizatidine on metronidazole resistant H. pylori strains could be explained by the effect on fumarate reductase, whereas the effect of omeprazole is different and could be an inhibition of a proton pump in H. pylori. Reversal of antimicrobial resistance with the help of different non-antibiotics seems to be possible by using quite different compounds, and is therefore to be explained by different molecular mechanisms. PMID- 11932142 TI - Prevalence of beta-lactamase production in H. influenzae isolated in Latin America in 1998-1999: results of the LASER study. AB - The objective of this study was to assess the prevalence of beta-lactamase production in Haemophilus influenzae clinical isolates obtained throughout Latin America and the West Indies in 1998-1999. Isolates were collected from 15 centres (seven countries), identified by standard methods and grouped by patient age. The overall prevalence of beta-lactamase production was 17.8% (270/1513 isolates). The prevalence of beta-lactamase positive strains varied between countries, with the highest prevalence detected in Panama (23.4%, 29/124) and the lowest in the West Indies (10.5%, 4/38). beta-Lactamase-positive strains were more frequently isolated from children aged < or =3 years (22.0%) and from adults aged > or =65 years (26.5%). The high prevalence of beta-lactamase production found should be considered when choosing empirical antibiotic therapy where H. influenzae is suspected. PMID- 11932143 TI - Prevalence of antimicrobial resistance in Haemophilus influenzae, Streptococcus pneumoniae, Moraxella catarrhalis and Streptococcus pyogenes: results of a multicentre study in Turkey. AB - The in vitro activities of several antimicrobial agents against clinical isolates of Streptococcus pneumoniae (283), Haemophilus influenzae (272), Moraxella catarrhalis (179) and Streptococcus pyogenes (256) were determined in a multicentre study with the participation of five hospitals from four cities in Turkey. Penicillin resistance in S. pneumoniae was evaluated using the E-test and the remaining agents by disk diffusion. For S. pneumoniae overall 25.8% of the isolates were intermediately and 3.9% were highly resistant to penicillin and resistance to chloramphenicol, azithromycin and trimethoprim/sulphamethoxazole (TMP/SMX) was 3.8, 2.1 and 55.4%, respectively. Seven percent of H. influenzae produced beta-lactamase and all were susceptible to cefotaxime and azithromycin; the highest rate of resistance, 23.5%, was for TMP/SMX. Eighty-one percent of M. catarrhalis isolates produced beta-lactamase, 18.4% were resistant to TMP/SMX and all were susceptible to sulbactam/ampicillin combination. Resistance to chloramphenicol and azithromycin of S. pyogenes was 2.2 and 1.9%, respectively. PMID- 11932144 TI - Prevalence of extended spectrum beta-lactamases among Enterobacteriaceae: an Italian survey. AB - The prevalence of extended-spectrum beta-lactamase (ESBL) production by consecutive non-repeated isolates of Enterobacteriaceae was determined over a 6 month period. A total of 8015 strains were isolated from ten Italian laboratories and 509 (6.3%) of these were designated ESBL producers from the results of a double-disk synergy test. Escherichia coli was the most isolated microrganism, followed by Klebsiella pneumoniae and Proteus mirabilis. Providencia stuartii (28.1%) was the most frequently isolated ESBL producer, followed by K. pneumoniae and Enterobacter aerogenes (20.5%). However, amongst all ESBL producers, K. pneumoniae (38.2%) was the most represented followed by P. mirabilis (25.7%). All the strains positive to DD tests were confirmed for the carriage of TEM and SHV genes using colony-blot hybridisation (CH). A total of 447 strains (88.0%) were CH-positive, of which 42.3% hybridised with the TEM-type probe, 30.1% with the SHV-type probe and 15.6% with both probes. In conclusion, our findings indicate that 6.3% of all Enterobacteriaceae tested produced ESBLs, 42.3% of which were TEM-derived enzymes. More than 20% of P. stuartii, K. pneumoniae and E. aerogenes harbour these enzymes. The double-disk test seems to be a useful test to identify ESBL producing strains. PMID- 11932145 TI - Antimicrobial susceptibility of Streptococcus species isolated from clinical mastitis in dairy cows. AB - The antimicrobial susceptibility was determined for 50 Streptococcus uberis, 42 S. dysgalactiae subsp. dysgalactiae and eight S. agalactiae strains isolated from cow mastitis. Only 27% of the strains were susceptible to all antimicrobial compounds tested. Resistance to tetracycline was most frequent (particularly for S. dysgalactiae strains), then macrolide and/or lincomycin resistance. High level resistance to streptomycin and kanamycin was detected. All S. dysgalactiae and S. agalactiae strains were susceptible to beta-lactams but 44% of the S. uberis strains showed an elevated penicillin G MIC. All strains were susceptible to chloramphenicol and rifampicin. PMID- 11932146 TI - Screening of acyl hydrazide proteinase inhibitors for antiparasitic activity against Trypanosoma brucei. AB - The major cysteine proteinase (brucipain) of Trypanosoma brucei is a target for chemotherapy of African Sleeping Sickness. We have screened a non-peptidyl acyl hydrazide proteinase inhibitor library of 500 compounds for inhibition of brucipain. Those 21 compounds with IC(50) values of <40 microM were tested for efficacy against bloodstream forms of T. brucei in cell culture. Eight acyl hydrazides showed 50% or more inhibition of trypanosome replication at <1 microM. The trypanocidal acitivity of the most effective compounds was comparable with those of the commercial antitrypanosomal drugs suramin and diminazene aceturate. However, these acyl hydrazides exhibited varying cytotoxicity towards human HL-60 cells and therefore, only less favourable selectivity indices compared with the commercially available drugs. Nevertheless, the data support the potential of acyl hydrazides as antitrypanosomal chemotherapeutic agents for treatment of sleeping sickness. PMID- 11932147 TI - Pharmacokinetics of teicoplanin in patients undergoing chronic haemodialysis. AB - In order to define a dose regimen of teicoplanin for patients undergoing chronic haemodialysis so that they achieved trough drug serum levels above 10 mg/l, two single doses of 5 and 10 mg/kg were administered intravenously in seven anuric patients immediately after the end of haemodialysis. Concentrations of teicoplanin were determined by a microbiological assay in samples collected from peripheral veins via the arterial and the venous lines of the fistulae and from the dialysate during haemodialysis. The administration of a 5 and 10 mg/kg dose gave mean C(max) of 62.80 and 122.43 mg/l, mean AUC of 526.43 and 1103.98 mg h/l, mean half life (t(1/2)) of 109.09 and 107.06 h, mean clearance rates of 12.85 and 12.44 ml/min, mean apparent volumes of distribution of 1.68 and 1.68 l/kg and mean volumes of distribution at steady state of 0.31 and 0.28 l/kg, respectively. Trough serum levels above 10 mg/l were found for 24 h after the administration of the 5 mg/kg dose and for 48 h after the administration of the 10 mg/kg dose. Teicoplanin was not detected in the dialysate. Its concentrations in both the arterial and the venous lines of the fistulae were similar. Based on the time period after the administration of teicoplanin where the desired trough serum levels were found and on the observed t(1/2), it is proposed that teicoplanin should be administered at a dose of 10 mg/kg at 48-72 h intervals, in patients undergoing chronic haemodialysis for the therapy of infections caused by Gram positive cocci. PMID- 11932148 TI - Multidrug resistance in bacterial isolates from blood cultures of haematology patients. AB - The bacteria most frequently isolated from blood cultures of haematology patients in Warsaw were staphylococci (58.0%), Enterobacteriaceae (18.6%), non-fermenting rods (6.9%), enterococci (4.3%) and anaerobes (4.3%). Coagulase-negative staphylococci were the most common species isolated (92.7%) with 83.2% of these strains resistant to methicillin. Among enteric bacteria, 17.3% strains produced extended-spectrum-beta-lactamases. All eight isolates of enterococci showed high level resistance to aminoglycosides. PMID- 11932149 TI - Antimicrobial resistance of Staphylococcus aureus isolated from skin infections. AB - The antimicrobial susceptibility of 229 strains of Staphylococcus aureus isolated from various skin infections was determined against 22 antimicrobial agents by the agar dilution method. The clinical isolates were most sensitive to vancomycin, teicoplanin, mupirocin and fusidic acid. No strains were resistant to vancomycin or teicoplanin. Three strains were highly resistant (MIC > or =100 mg/l) to mupirocin and eight strains to fusidic acid. The MIC(50) of all antimicrobials, except for gentamicin, were below 3.13 mg/l. The incidence of resistance to penicillin, cephalosporins and clindamycin ranged from 20 to 30%. The occurrence of gentamicin, erythromycin and roxithromycin resistance was high at 55.2, 39.6 and 39.1%, respectively. Methicillin resistance occurred in 21.0% of strains. The incidence of organisms with MIC > or =3.13 mg/l to oxacillin was 24.3%. These results were comparable to the average rate of MRSA in Japanese dermatological specimens. PMID- 11932151 TI - Increase of in vitro amoxycillin bactericidal activity by clavulanic acid against Neisseria meningitis using time-kill curves. PMID- 11932150 TI - Open-label, multicentre, emergency-use study of clinafloxacin (CI-960) in the treatment of patients with serious life-threatening infections. AB - In an open-label emergency-use study, 23 patients with bacterial infections caused by multiply drug resistant pathogens were treated with clinafloxacin. Efficacy and safety were evaluated by tabulating investigators' assessments at the end of treatment, treatment discontinuations and adverse event data. Most of the patients were seriously ill and had multi-organ infections, primarily respiratory tract infections such as nosocomial pneumonia and gastrointestinal infections. Eleven patients were successfully treated, two had treatment failure and 10 were not evaluable because the patients died of their underlying disease. Considering that most of the patients had several infections caused by multiply resistant pathogens, clinafloxacin may be useful for the treatment of such life threatening episodes. PMID- 11932152 TI - Evolution of HIV disease in the third millennium: clinical and related economic issues. PMID- 11932153 TI - Can pneumonectomy for non-small cell lung cancer be avoided? An audit of parenchymal sparing lung surgery. AB - BACKGROUND: Lung cancer resection rates are suboptimal in the UK. Pneumonectomy has a higher perioperative mortality risk than lobectomy. To increase resection rates and improve outcomes we have implemented a policy of parenchymal sparing surgery for tumours involving a main stem bronchus. METHODS: In a prospective 4 year study of 119 consecutive patients operated upon by a single surgeon the perioperative course, pathology and survival were compared for 81 patients undergoing pneumonectomy and 38 patients in whom pneumonectomy was avoided by bronchoplastic+/-angioplastic procedures. RESULTS: The rate of pneumonectomy decreased significantly with increasing experience with parenchymal sparing surgery (R(2)=0.98, P<0.001) with 21 of the last 30 patients (70%) avoiding pneumonectomy. There were no significant inter-group differences in patient characteristics, perioperative course or outcome. One-year survival was 64% after pneumonectomy and 73% after sleeve lobectomy. However the perioperative loss of respiratory function was significantly lower in the patients in whom pneumonectomy was avoided (P=0.0003). CONCLUSIONS: Pneumonectomy can be avoided in a large proportion of patients with non-small cell lung cancer of a main stem bronchus without adversely affecting outcome but with preservation of lung function PMID- 11932154 TI - Microscopic (R1) and macroscopic (R2) residual disease in patients with resected non-small cell lung cancer. AB - OBJECTIVES: This retrospective study evaluates the probability of survival in patients who had undergone resection for non-small cell lung cancer (NSCLC) and in whom residual disease at the resection margins was found. METHODS: During a period of 6 years, 596 patients with NSCLC were operated upon with curative intention. Residual disease at the resection margin was divided into microscopic (R1) and macroscopic (R2). RESULTS: Twenty-six patients (4.4%) showed R1 and 12 (2%) R2 residual disease. An extrabronchial (thoracic wall, vessels) R1 situation was found in five patients and a bronchial R1 infiltration in 21 cases. The bronchial resection margin was subject to peribronchial infiltration in most cases (16/21). A total of 17/21 (65%) patients with bronchial infiltration had N2 disease. Thirty day lethality was 3.8% in the R1 group. Fifteen patients had postoperative irradiation. The 5-year survival rate for patients with R1 resection was 14%. The differences in survival between patients with extrabronchial vs. bronchial infiltration and N0/N1 vs. N2 were significant using univariate analysis. Adjuvant radiation did not result (especially in N2 disease) in a survival benefit. Among 12 patients with macroscopic residual disease (R2), 3/12 (25%) died within the first 30 days after the operation, and none of the R2 patients survived the first year after the operation. CONCLUSIONS: Patients with an R1 situation have a survival rate of 14% comparable to curative resected patients (RO) in stage III. Adjuvant radiation had no clear effect on survival. Patients with macroscopic tumor (R2) should receive palliative treatment after the operation depending on their condition. PMID- 11932155 TI - Pulmonary metastatic melanoma -- the survival benefit associated with positron emission tomography scanning. AB - OBJECTIVE: Positron emission tomography (PET) scanning is more sensitive at detecting metastatic disease than conventional radiological techniques. For patients with pulmonary metastatic melanoma, we investigate if PET scanning to detect occult extra pulmonary disease prior to thoracotomy and metastectomy is associated with improved survival compared to patients staged by conventional radiology. METHODS: Between November 1984 and December 1999, 121 patients (90 males, 31 females) have undergone a thoracotomy and pulmonary metastectomy for metastatic melanoma. The age range was 19-84 years (mean 57, median 59). In every case all palpable nodules were removed and the diagnosis confirmed histologically. A total of 68 (56%) patients had a PET scan preoperatively, 53 (44%) underwent conventional or nuclear imaging. Patients with only radiologically isolated pulmonary disease are included. RESULTS: Survival is 100% complete and totals 238 pt/years (mean 2.2 years, median 1.4 years). Survival (+/ SE) at 1, 3, 5 and 7 years for all patients is 68% (+/-4.5) (n=67), 36.6% (+/ 5.2) (n=27), 22.1% (+/-4.8) (n=15) and 13.5% (+/-4.2) (n=7), respectively. Survival (+/-SE) was significantly better at 3 and 5 years in patients who underwent a PET scan preoperatively (Log rank P=0.002). There was no significant difference in survival by 7 years. CONCLUSIONS: There is a significant survival benefit associated with excluding extra pulmonary disease using a PET scan prior to thoracotomy and metastectomy. We recommend that PET scanning be used in the investigation of patients with pulmonary metastatic melanoma prior to metastectomy. PMID- 11932156 TI - The effect of subcutaneous tumour implantation in a murine lung tumour model. AB - OBJECTIVES: It was hypothesized that if tumour were implanted subcutaneously within a Millipore Chamber (MPC), then this would result in an 'anti-tumour' immune response. Such an approach could have potential as an adjuvant tumour therapy when combined with surgical resection. A murine lung tumour model was used to test this hypothesis. METHODS: Lung tumours were induced in 245 syngeneic mice by intraperitoneal 4-[methylnitrosamino]-1-[3 pyridyl]-1-butanone. In addition, MPCs were implanted containing either normal lung (Group A) or lung tumour (Group B). Group C had no implanted MPCs. These animals were sacrificed between 1 and 8 weeks following implantation and the stage of lung tumour development as assessed by the surface tumour count (STC) of their left lungs was compared between the different groups. The presence of CD4(+) and CD8(+) T cells in the local reactions surrounding the implanted chambers was also compared between Groups A and B at 1 week post-implantation. RESULTS: At 1 week, the STC was significantly lower in Group B (2.4+/-0.6) than in both Groups A (4.7+/-0.6) and C (4.9+/-0.9; P=0.02). In addition, at 1 week, there was a significantly greater proportion (%) of CD4(+) cells in the local reactions of Group B (52+/-3) than in Group A (35+/-3; P=0.001). Between 2 and 8 weeks post-implantation, there were no further significant differences in tumour development between the groups. CONCLUSIONS: Although the findings of an 'early' response were consistent with the hypothesized benefit of tumour implantation within MPCs, the later results have not confirmed its potential as an adjuvant therapy. PMID- 11932157 TI - Does lung biopsy help patients with interstitial lung disease? AB - OBJECTIVES: The decision to perform lung biopsy in the evaluation of interstitial lung disease (ILD) is based on the probability that this examination will yield a specific diagnosis, leading to a change in treatment. The purpose of this study was to identify factors that influence the diagnostic yield of lung biopsy for ILD. METHODS: One hundred patients underwent lung biopsy for ILD over a 5-year period. There were 59 men and 41 women; with a median age of 51.5 years. Thirty percent underwent open lung biopsy, while 70% had videothoracoscopic biopsy. Patient and disease characteristics, prior diagnostic studies, pre-operative therapy, biopsy type, site, size, number, and laterality were compared to identify factors that might influence diagnostic yield. RESULTS: Forty-two percent had a specific diagnosis, while 58% had a non-specific diagnosis. Right side was selected in 57.1% of patients with a specific diagnosis and 48.3% of patients without a specific diagnosis (P=0.381). Right lower lobe was the main site for biopsy in the specific diagnosis group compared to the non-specific group (35.7 versus 20.7%, P=0.095). Left upper lobe was the main site for biopsy in the non-specific diagnosis group compared to the specific diagnosis group (41.4 versus 23.8%, P=0.067). Mean volume of biopsy was 12.3 cm(3) in the specific diagnosis group and 12 cm(3) in the non-specific diagnosis group (P=0.373). Two or more biopsies were carried out in 38.1% of the specific diagnosis group compared to 25.9% of the non-specific diagnosis group (P=0.192). There were no significant factors in predicting a diagnostic yield. Of those patients with a specific diagnosis, 59.5% had therapy altered, compared to 55.2% of those with a non-specific diagnosis (P=0.664). CONCLUSIONS: Lung biopsy does not always provide a specific diagnosis and does not always change therapy. The site, size, number, and laterality of the biopsy specimen have no definite influence on diagnosis. There is a trend to improve diagnostic yield by carrying out two or more biopsies on the right lung. PMID- 11932158 TI - Long-term outcome of staged versus one-stage bilateral thoracoscopic reduction pneumoplasty. AB - OBJECTIVE: In a prospective non-randomized study, we tested the hypothesis that unilateral reduction pneumoplasty followed by completion of bilateral treatment at the reappearance of symptoms might result in more sustained improvements and better survival than one-stage bilateral treatment. METHOD: Fifty-nine patients undergoing bilateral thoracoscopic reduction pneumoplasty as a one-stage (n=33) or staged (n=26) procedure were evaluated on. The main indication for staged reduction pneumoplasty was symptom deterioration after unilateral treatment for asymmetric emphysema. Complete clinical assessment was carried out preoperatively and every 6 months postoperatively. RESULTS: The mean length of follow-up was 34+/-15 months. Interval time between operations in the staged group averaged 15.2 months. There was no inter-group difference in baseline data. Peak improvements in forced expiratory volume in 1 s (FEV(1)), forced vital capacity (FVC) and residual volume (RV) was significantly greater following one-stage bilateral reduction pneumoplasty. In particular, Delta FEV(1) was 0.33+/-0.2 l in the staged group and 0.43+/-0.2 l in the one-stage group (P=0.007). At 48 months, FEV(1), RV and 6-min-walking-test (6MWT) were still significantly improved only in the staged group. Four-year survival was 70% in the staged group and 81% in the one-stage group (Cox-Mantel test, P=not significant). CONCLUSION: Durable physiological improvements and satisfactory survival were achieved in this study for up to 4 years following either staged or one-stage bilateral reduction pneumoplasty using thoracoscopic technique. However, while peak improvements in FEV(1), FVC and RV were significantly greater following one-stage bilateral reduction, long-term improvements in FVC and 6MWT were more stable following a staged procedure. We speculate that sequential unilateral reduction pneumoplasty may reduce the mechanical stress in the lung leading to less steep postoperative deterioration of respiratory function. PMID- 11932159 TI - Surgical treatment in bronchiectasis: analysis of 166 patients. AB - BACKGROUND: Bronchiectasis is usually caused by pulmonary infections and bronchial obstruction. It is still a serious problem in developing countries as our country. We reviewed the morbidity and mortality rates and outcome of surgical treatment for bronchiectasis. PATIENTS AND METHODS: Between 1990 and 2000, 166 patients (92 female and 74 male patients) underwent pulmonary resection for bronchiectasis. The mean age was 34.1 years (range, 7-70 years). Mean duration of symptoms was 5.7 years. RESULTS: Symptoms were copious amount of purulent sputum in 135 patients, expectoration of foul-smelling sputum in 109, hemoptysis in 35 and cough in all patients. The indication for pulmonary resection was failure of medical therapy in 158 patients, massive hemoptysis in five and lung abscess in three. The disease was bilateral in six patients and mainly confined to the lower lobe in 127. One hundred and twenty patients had a lobectomy, 13 had a pneumonectomy, 21 had a segmentectomy and a combination of these approaches in 18. Operative morbidity and mortality were seen in 18 (10.5%) and in three (1.7%) patients, respectively. Follow-up was complete in 148 patients with a mean of 4.2 years. Overall, 111 patients were asymptomatic after surgical treatment, symptoms were improved in 31, and unchanged or worse in six. CONCLUSIONS: Surgical treatment of bronchiectasis is more effective in patient with localized disease. It is satisfactory with acceptable ratio of morbidity and mortality. PMID- 11932160 TI - Outcome of acid ingestion related aspiration pneumonia. AB - BACKGROUND: The objective of this study is to assess the incidence and long-term results of a rarely discussed medical problem -- aspiration pneumonia resulting from the intentional ingestion of acid. MATERIALS AND METHODS: The medical records of 370 patients treated at one tertiary care institution for corrosive acid injury during a 12-year period were reviewed retrospectively. The study subjects included any patients who were found to have acid ingestion related aspiration pneumonia confirmed by chest film within 24h of injury. All available data of these patients with or without aspiration pneumonia were analyzed. RESULTS: Of the 370 patients with corrosive acid injury, 15 (4.2%) had acid aspiration pneumonia which was related to their intentional ingestion of a strong acid, hydrochloric acid (pH<1). The data for 14 patients with aspiration pneumonia and 268 without aspiration pneumonia was complete and available for analysis. Patients with aspiration pneumonia were found to be significantly older (52.2+/-6.2 to 41.7+/-0.9 years old, P=0.017), had a higher incidence of nasogastric tube irrigation (35.7-6.0%, P=0.000), had more conscious disturbance (50.0-17.5%, P=0.016), and required more endotracheal tube intubation (50.0-3.0%, P=0.000). Aspiration pneumonia was found to significantly increase the mortality rate in acid injured patients who required emergency abdominal surgery (87.5 32.0%, P=0.000) and in those who did not (28.5-5.1%, P=0.05). Two of the six survivors of aspiration pneumonia later developed laryngeal sequelae. CONCLUSIONS: Aspiration pneumonia rarely occurs as a consequence of acid ingestion. When it does occur, it greatly increases the mortality rate of those involved. For those who survive, physicians can expect some laryngotracheal sequel in long-term follow-up. PMID- 11932161 TI - Prevention of recurrent empyema after pneumonectomy for chronic infection. AB - OBJECTIVES: Pneumonectomy in chronic pulmonary infection with empyema is associated with a high mortality rate and an increased risk of recurrent empyema. The surgical resection is technically demanding, and successful management continues to be a challenge. METHODS: We evaluated a concept which combines (pleuro-)pneumonectomy or completion pneumonectomy with surgical debridement of the pleural cavity and packing with povidine-iodine soaked dressings. The debridement and packing is repeated in the operating theater after 48 h until the chest cavity is macroscopically clean. Finally, the pleural space is obliterated with antibiotic solution. RESULTS: Between February 1997 and October 2000, 11 patients (average age of 59 years, ranging from 25 to 84) with destroyed lung caused by tuberculosis (six), aspergilloma (two), bronchiectasis (one), esophago pleural fistula (one) or broncho-pleural fistula after lobectomy for bronchial carcinoma (one) and ongoing chronic infection with acute empyema (ten) (25-2500 days between first and definitive therapy) were treated. Pleural culture findings showed Aspergillus in four, Mycobacterium in two, Enterococcus in two, Candida in one and Staphylococcus in one, respectively. The mean number of interventions was 2.9 (2-4). The chest was definitively closed in all patients within 1 week. The mean hospitalization time was 19 days (9-31 days). In the follow-up (10-54 months), there was no recurrence of empyema. One patient (84 years) died at day 31, due to sepsis. CONCLUSIONS: Pneumonectomy combined with repeated surgical debridement and antimicrobial therapy enables the successful treatment of chronic pulmonary infection with empyema within a short time period. PMID- 11932163 TI - Management of esophageal foreign bodies: a retrospective review of 400 cases. AB - OBJECTIVE: A retrospective study was conducted in 400 patients with esophageal foreign bodies (EFB) to assess characteristics of EFB and methods of treatment. METHODS: From 1962 through 1998, 400 patients with EFB were treated in our department on an emergency basis. There were 202 men (50%) and 198 women (49.6%) ranging in age from 1.5 to 95 years. The main symptoms patients complained of were difficulty in swallowing and pain. Detailed anamnesis, oropharynx and hypopharynx examination and finally radiological examination were the diagnostic tools. The location of the FB was in the cervical esophagus in 57% of cases, in the thoracic one in 26% and at the cardioesophageal junction in 17%. The most common objects found were bones, morsels, coins and needles. RESULTS: The treatment consisted of rigid esophagoscopy under general anesthesia in 343 (85.7%) of our cases. In 57 cases (14.3%) other means such as flexible esophagoscopy, Fogarty or Foley catheters and bougienage turned to be very useful. Only 12 patients (3%) were led to surgery because either extraction was impossible or perforation was present. No major complications occurred in the surgical group, whereas in the group of rigid esophagoscopy, there was one iatrogenous esophageal perforation that presented with empyema thoracis successfully treated. Finally, there was a case of an aortoesophageal fistula with mortal outcome perioperatively. CONCLUSIONS: (1) Esophagoscopy is a reliable method in the treatment of EFB impaction. (2) Alternative methods such as bougienage, etc., can be used only in selected cases with smooth foreign bodies. (3) Surgical treatment is unavoidable in cases of irretrievable EFB or esophageal perforation. PMID- 11932162 TI - Iatrogenic ruptures of the tracheobronchial tree. AB - OBJECTIVE: Iatrogenic tracheobronchial ruptures are seldom but severe complications after intubation or bronchoscopy. Therefore, we evaluated the reasons, the subsequent therapy and the outcome of patients with tracheal rupture, who were admitted to our hospital. METHODS: In a retrospective study we examined 19 patients (15 women, four men; 43-87 years) treated for acute tracheobronchial lesions. Eleven (58%) patients had a tracheobronchial rupture by single-lumen tube, four (21%) by double-lumen tube and two patients (10%) by tracheal cannula. A total of 47% of whom were carried out under emergency conditions. Two patients had a rupture due to a stiff bronchoscopy. Mean symptoms were mediastinal and subcutaneous emphysema. Two emergency collar incisions had been done. RESULTS: The localization of ruptures was in all cases in the paries membranaceus, length: 1-7 cm (mean: 4.8 cm). The interval between the onset of symptoms and the diagnose differed widely (up to 72 h), nine (47%) diagnoses were made during intubation/bronchoscopy. One patient, with a small tear (1 cm) was treated conservatively with fibrin-glue. The other 18 patients had surgical repair through a thoracotomy. The postoperative mortality was determined with 42%, which was not dependent on the rupture but basically by the underlying diseases requiring intubation. CONCLUSIONS: Iatrogenic tracheal rupture is a dangerous complication with potentially high postoperative mortality, mostly influenced by the underlying disease. Early surgical repair must be the preferred treatment. PMID- 11932164 TI - Predictive factors for response to neoadjuvant therapy in patients with oesophageal cancer. AB - BACKGROUND: Preoperative radio-chemotherapy (RCX) was introduced to improve the outcome of patients with oesophageal cancer (EC), but conflicting results have been released. Some 20-30% of patients show a complete pathological response, however, the perioperative morbidity and mortality is increased. To search for factors indicating response prior to the onset of RCX we investigated the proliferative activity (MIB-1), the expression of vascular endothelial growth factor (VEGF), and the capillary density (CD34) in samples of EC obtained by endoscopy prior to the start of the treatment. METHODS: Forty-six (MIB-1) and 21 (VEGF, CD34) tissue specimens of ECs were available from 56 patients undergoing pretherapeutic endoscopy, RCX and surgery. Perioperative morbidity was divided into surgery and non-surgery related morbidity. MIB-1, VEGF and CD34 expression were investigated immunohistochemically. Multivariate analysis was carried out to prove independence of investigated variables. RESULTS: Postoperative morbidity was noticed in 54 of 56 operated patients. Eight of 56 patients who received RCX died in hospital. Survival was significantly different between the group of complete responders (n=14) and non-responders (n=23; P=0.0026). None of the investigated tumour samples from patients with a complete response (CR) had a proliferation index of less than 45. Tumour samples from patients with a CR showed a VEGF expression of 10.7 compared with 36.58 of tumours with no response (P=0.035). CD34 expression showed a correlation with VEGF expression. The relation of mean indices of VEGF expression and proliferative activity in tumours from patients with complete, partial or no response was 10.7:58.8, 18.3:53.8 and 36.6:43.5, respectively. CONCLUSIONS: According to these results, it may be expected that tumours with a VEGF/MIB-1 ratio of 1:6 or less prior to RCX will respond to this therapy. PMID- 11932166 TI - Vocal cord paralysis after open-heart surgery. AB - Vocal cord paralysis is a known entity often described as a complication of neck surgery. A less frequent site of injury to the recurrent laryngeal nerve is the chest. The left side is usually more affected than the right side in view of its long intrathoracic segment. Only few cases of right vocal cord paralysis following open-heart surgery are reported in the literature. The purpose of this article is to review the common possible mechanisms of injury to the right recurrent laryngeal nerve following open-heart surgery in order to draw the attention of the caring physician to the clinical significance of such a complication. In fact, transient hoarseness following open-heart surgery may be an ominous sign of recurrent laryngeal nerve injury. It should not be assumed to be secondary to intralaryngeal edema. Several mechanisms of injury to the recurrent laryngeal nerve have been suggested: (1) through central venous catheterization; (2) by traction on the esophagus; (3) by direct vocal cord damage or palsy from a traumatic endotracheal intubation; (4) trauma by compression of the recurrent laryngeal nerve or its anterior branch at the tracheoesophageal groove by an inappropriately sized endotracheal tube cuff; (5) by a faulty insertion of a nasogastric tube; (6) median sternotomy and/or sternal traction pulling laterally on both subclavian arteries; (7) direct manipulation and retraction of the heart during open-heart procedures; (8) hypothermic injury with ice/slush. If vocal cord paralysis was overlooked as a possible complication of open-heart surgery, the patient may suffer from dysphonia in addition to problems of paramount importance such as inefficient cough and aspiration. Although it is true that the incidence of vocal cord paralysis remains very low, yet its presence is alarming and necessitates close follow up on the patient for the possible need of surgical intervention if recovery fails. PMID- 11932165 TI - Thirteen years follow-up after radical transsternal thymectomy for myasthenia gravis. Do short-term results predict long-term outcome? AB - OBJECTIVE: Long-term evaluation of efficacy and quality of life after radical surgical approach for myasthenia gravis (MG). Comparison between short-term follow-up and long-term outcome. METHODS: All patients (n=26, 16 men and 10 women, mean age: 40.7 years) underwent total transsternal thymectomy for MG between 1986 and 1989. Prospective analysis of the patients for short-term follow up (mean 22.4 months) was published in 1991. The same group of patients was reevaluated in 2001 (range of follow-up 11.4-15.2 years) and assessed according to the classification of Osserman and Oosterhuis. RESULTS: Mean follow-up was 13.0 years (range 11.4-15.2 years). Two patients were lost from follow-up and one died 4 years after thymectomy for reasons unrelated to MG (n=23). No early or late postoperative mortality was observed. One sternal osteomyelitis occurred. Late postoperative morbidity included sternal instabilities (n=2), mild residual thoracic pain (n=6), and hypertrophic scars (n=7). Five patients were rehospitalized for aggravating MG and needed plasmapheresis (n=3) and intubation (n=1). Thirteen patients (56.5%) showed objective clinical improvement, including six patients (26.1%) with complete remission. Eleven patients (47.8%) do not take any medication at all. Because some late relapse may occur several years after operation, the rate of improvement decreased slightly, whereas the difference between short and long-term follow-up was not statistically significant (P=0.405). Twenty patients (87%) returned to work, including part-time occupation (n=4). Fourteen patients (61%) are performing sports regularly. CONCLUSIONS: Our data confirm that radical, transsternal thymectomy is an effective and safe therapeutic modality for MG. Short-term results seem to deteriorate over time, therefore long-term studies for minimally invasive approaches have to prove equal results before replacing the standard procedure. PMID- 11932167 TI - Medial degeneration does not involve uniformly the whole ascending aorta: morphological, biochemical and clinical correlations. AB - OBJECTIVE: To investigate whether and how the severity of medial degeneration (MD) lesions varies along the circumference of the dilated intrapericardial aorta. METHODS: Two groups of aortic wall specimens, respectively harvested 1cm distal to the non-coronary (NC) sinus (right postero-lateral wall) and to the right coronary sinus (anterior wall) in 22 patients undergoing surgery for dilatation of the intrapericardial aorta associated with aortic valve disease, were separately sent for pathology, morphometry and ultrastructural examination. MD lesions found at histology were classified into three degrees of severity. MD mean degree and morphometric findings in postero-lateral ('NC') and anterior ('coronary') specimens were compared by paired t-test. Correlation between degree of aortic dilatation at echocardiography and severity of MD was assessed separately for each of the two groups of specimens. After the preliminary results of the morphological study, we decided to send the specimens for biochemical investigation of protein electrophoretic patterns. This was performed in the last seven patients of this series. RESULTS: At histology, MD was found in all cases. A higher mean MD degree was found in the NC group (2.59+/-0.50 versus 1.59+/-0.67 in the coronary group; P<0.001). At morphometry, normal smooth muscle cells in the NC specimens were significantly reduced (P=0.012) and the length (P=0.011) and number (P=0.015) of elastic fibres reduced and increased, respectively. Correlation between aortic ratio and MD degree was significant in the NC specimens (P<0.001), not in the coronary ones (P=0.227). Quantitative differences between coronary and NC proteins from the same patient and between coronary proteins from different patients were found at electrophoresis. However, at this stage of the study, the sample was too small to allow for the identification of proteins involved in those differences. CONCLUSIONS: MD lesions in dilated intrapericardial aorta are more severe in the right postero-lateral wall area, likely due to haemodynamic stress asymmetry. PMID- 11932168 TI - Evolution of allograft aortic valve replacement over 13 years: results of 275 procedures. AB - OBJECTIVE: We describe our center's experience with the use of allografts for aortic valve or root replacement, illustrating the impact on outcome of the changes made in surgical and preservation techniques. METHODS: Between 4/1987 and 1/2001 275 allografts were used in 267 consecutive patients to replace the aortic valve or root. All patients were prospectively followed over time. Mean patient age was 46 years (SD 16; range 0.06-83), male/female ratio was 201/74. Prior cardiac operations took place in 73 patients; 49 patients presented with active endocarditis. Pre-operative NYHA-class was III in 51%. Initially, the subcoronary technique was used (SC; N=95) while in recent years root replacement (ARR; N=180) became the technique of choice. Seven fresh (two pulmonary and five aortic) and 268 cryopreserved (four pulmonary and 264 aortic; 35 glycerol and 233 DMSO) allografts were implanted. Concomitant procedures took place in 133 (48%). RESULTS: Operative mortality was 5.5% (N=15) and during follow-up (99% complete) 29 more patients died. Overall cumulative survival was 73% (95% CI 65-81%) at 9 years postoperative and significantly better for SC compared to ARR patients (P=0.005). Freedom from allograft-related reoperation (N=34) was 77% (95% CI 69 85) at 9 years, and worse in the SC compared to ARR group due to increased early technical failure (P=0.03). Freedom from reoperation for structural valve deterioration (SVD; N=22) was 81% (95% CI 73-89) at 9 years and did not differ between SC and ARR (P=0.51). Independent predictors of degenerative SVD were younger patient age (HR 0.93 with age as continuous variable; 95% CI 0.90-0.97), older donor age (HR 1.06 with age as a continuous variable; 95% CI 1.00-1.11), larger allograft diameter (HR 1.38; 95% CI 1.11-1.71) and the use of pulmonary allografts (HR 10.72; 95% CI 3.88-29.63). Calculated median time to reoperation for structural valve deterioration ranged from 23 years in a 65-year-old patient to 12 years in a 25-year-old. CONCLUSIONS: Aortic valve replacement with allografts yields adequate midterm results. Although important changes have been made over the years to improve durability, allografts still have a limited life span especially in young patients. PMID- 11932169 TI - Comparison of exercise and dobutamine echocardiography in the haemodynamic assessment of small size mechanical aortic valve prostheses. AB - OBJECTIVE: Doppler echocardiographic evaluation of prosthetic heart valve function is usually performed at rest although this situation is not representative of patients' daily activities. Following aortic valve replacement, patients most likely to remain symptomatic are those with a small aortic root and dobutamine or exercise echocardiography has been proposed to elicit the presence of abnormal haemodynamics or persistently elevated transvalvular gradients in these patients. This study was carried out to compare dobutamine echocardiography with a symptom limited treadmill exercise echocardiography in patients following aortic valve replacement with a small size (19 mm) St. Jude Mechanical valve prosthesis. METHODS: The study population consisted of ten unselected patients following aortic valve replacement. Dobutamine was infused intravenously starting at 5 microg/kg/min and increasing by 5 microg/kg/min at 15 min interval up to 20 microg/kg/min. Heart rate, blood pressure, cardiac output (CO), peak and mean gradients as well as effective orifice area (EOA) were measured. These parameters were also measured following a symptom limited treadmill exercise. RESULTS: Dobutamine stress increased heart rate (HR) and CO by 50 and 74%, respectively (both P<0.0002), and mean transvalvular gradient from 22+/-4.1 mmHg at rest to 40.0+/-10 mmHg at maximum stress (P<0.001). With exercise, HR and CO increased by 48 and 70%, respectively while mean transvalvular gradient increased from 22+/ 3.1 mmHg at rest to 38.0+/-6.4 mmHg (P<0.0001). The maximum increase in HR, CO and mean transvalvular gradient with dobutamine and exercise were similar however. There was no significant change in the EOA with either dobutamine or exercise. CONCLUSION: The result suggests that both treadmill exercise and dobutamine stress echocardiography are equally effective for the hemodynamic evaluation of small aortic valve prosthesis. PMID- 11932170 TI - Aortic valve replacement with the Toronto SPV: long-term clinical and hemodynamic results. AB - OBJECTIVE: Long-term durability and hemodynamics of stentless valves are unknown. Therefore, this study was undertaken to evaluate long-term clinical and echocardiographic outcome after aortic valve replacement with the Toronto stentless porcine valve (SPV) bioprosthesis at our institution. METHODS: Between 1991 and 1998, the Toronto SPV was implanted in 255 patients (mean age, 63+/-11 years, range 22-83 years, 181 males and 74 females). Preoperative diagnoses were aortic stenosis (76%), aortic insufficiency (12%) and mixed lesion (12%). New York Heart Association class III and IV were present in 49% (126/255) of the patients preoperatively. Mean valve size implanted was 26.5+/-2.1mm and in addition 34% (86/255) of the patients had coronary artery bypass surgery. RESULTS: Early mortality was 0.8% (2/255). Actuarial survival at 7 years was 90+/ 3%. At 7 years, the freedom from cardiac death was 98+/-2%; from valve-related death, 99+/-1%; from valve reoperation, 97+/-2%; from structural valve degeneration, 97+/-2%; from thromboembolism, 95+/-2%; and from endocarditis, 99+/ 1%. At 7 years of follow-up, the transvalvular peak and mean pressure differences across the aortic valve measured with Doppler echocardiography was 9.6+/-5.1 and 3.6+/-2.0 mmHg, respectively. CONCLUSIONS: The Toronto SPV has provided excellent clinical and hemodynamic results up until 7 years of follow-up. PMID- 11932171 TI - Bovine valved venous xenografts for RVOT reconstruction: results after 71 implantations. AB - BACKGROUND: Pediatric right ventricular outflow tract (RVOT) reconstruction with homo- or porcine xenografts is problematic because of limited availability, lack of material for reconstruction, early degeneration, and tissue ingrowth. Contegra, a bovine jugular vein graft, might be an interesting alternative to overcome these problems. PATIENTS AND METHODS: Within a Federal Drug Administration controlled study, we implanted 71 Contegra pulmonary valved conduits from May 1999 to September 2001 in 71 patients (male/female 33/38) in the age range 2 days-17.4 years, median 1.2 years. Twenty five were primary repairs, 22 had previous graft implantations, and 24 had other repairs/palliations. Preoperative diagnoses: truncus arteriosus communis (19 patients), tetralogy of Fallot (32), double outlet right ventricle (13), transposition of the great arteries (5), and two rare complex malformations. The size of implanted Contegra conduits ranged from 12 to 22 mm. Echocardiography was performed at 1 and 3 months, and then every 3 months postoperatively. Follow-up time was 27 months (maximal), 80 years in total. Results were compared with our 52 homograft- and 30 Tissuemed porcine xenograft recipients. RESULTS: Contegra enables the surgeon to perform all anastomoses without additional material. Its tissue is very apt for suturing and its insufficiencies are common, but without clinical significance or tendency to increase. We saw no sign of conduit or valve degeneration during the whole follow-up up to 27 months. There were no device related adverse events. Redos: five for peripheral pulmonary arteries, two residual ventricular septum defect (VSD) closures. There were six deaths (five early, one late). The maximal transvalvular gradients of 25-42 mmHg were measured in seven patients; these gradients did not increase further during the follow-up. Six patients with completely intact Contegra conduits developed pressure gradients of more than 70 mmHg immediately distal from the conduit. At 27 months, Contegra grafts were advantageous compared to homografts with respect to survival and freedom from explantation. Right ventricle to left ventricle (RV/LV) ratio development and freedom from explantation/redo were equal for Contegra conduits and homografts. Porcine Tissuemed xenografts were significantly inferior. CONCLUSION: The Contegra conduit offers unique tailoring and suturing options for primary and redo RVOT reconstruction. At 27 months, its durability seems at least equivalent to homografts and is superior to porcine Tissuemed xenografts. PMID- 11932172 TI - Intermediate results with correction of tetralogy of Fallot with absent pulmonary valve using a new approach. AB - OBJECTIVE: The intermediate results achieved with a new technique for primary repair of tetralogy of Fallot with absent pulmonary valve syndrome are reported. METHOD: Apart from correction of tetralogy of Fallot, this approach includes translocation of the pulmonary artery anterior to the aorta and away from the tracheobronchial tree. Since November 1998 this technique has been employed in three symptomatic newborns and three infants with a diagnosis of tetralogy of Fallot with absent pulmonary valve. The median age at surgery was 37 days (range 14-256 days). The median weight at operation was 3.4 kg (range 2.9-4 kg). All patients had severe respiratory problems and congestive heart failure with cyanosis. RESULTS: There was no early or late death during the follow-up (median 27 months). One patient required redo due to failure to thrive as a consequence of right ventricle volume overload. Valved conduit was inserted to pulmonary position. Respiratory symptoms disappeared or were significantly reduced in all patients. Postoperative computed tomographic scan showed no compression of trachea and main bronchi, pulmonary artery was away from tracheobronchial tree in all patients. Patients are doing well with adequate growth. CONCLUSIONS: The new technique described here has a potential to reduce or eliminate bronchial compression by pulmonary artery. Translocation of pulmonary artery anterior to the aorta takes the dilated pulmonary artery away from the trachea and bronchial tree. This approach was found to be technically feasible and can be useful especially in symptomatic newborns and infants. PMID- 11932173 TI - Stage I palliation for hypoplastic left heart syndrome in low birth weight neonates: can we justify it? AB - OBJECTIVE: Although the outcome of cardiac surgery in neonates with low birth weight (LBW) has improved, LBW remains a risk factor for surgical palliation. Few surgical series of LBW patients include those with hypoplastic left heart syndrome (HLHS). To identify variables associated with poor outcome in this group, we reviewed our experience with patients with HLHS and LBW who underwent Stage I Norwood palliation. METHODS: Between January 1998 and December 2000, 20 consecutive LBW (<2500 g) neonates with HLHS (n=13) or HLHS variant (n=7) underwent surgical palliation. Retrospective review of all patient data and analysis to identify risk factors was performed. RESULTS: Mean age at surgery was 5.1+/-4.6 days (range 1-17), mean weight was 1.98+/-0.44 kg (range 1.1-2.5), including nine patients under 2 kg. Ten patients were born at <35 weeks gestation. Anatomic diagnosis included HLHS in 13 patients (10 with aortic atresia), unbalanced atrioventricular canal defect in two, double outlet right ventricle in two and other variants in three. Mean ascending aortic size was 4.0+/-1.8 mm (range 1.5-8). Associated cardiac defects were present in three patients, and a genetic syndrome and/or congenital anomaly was present in four of them. Mean circulatory arrest time was 60+/-10 min. Extracorporeal support was used perioperatively in 10 patients. Early mortality was 9/20 (45%). At a mean follow up at 22+/-10 months (range 8-38), six patients underwent stage II, and are awaiting stage III; four patients have completed their Fontan. Anatomic variant, ascending aortic size, prematurity, age at surgery, weight, duration of circulatory arrest, cardiopulmonary bypass time and associated non-cardiac anomalies were not risk factors for poor outcome whereas restrictive pulmonary venous drainage and coronary artery anomalies were associated with decreased survival. CONCLUSION: LBW newborns with HLHS and physiologic variants have an increased early surgical risk but have acceptable intermediate survival rates for subsequent palliation including Fontan. LBW and prematurity should not be contraindications to early surgical palliation. PMID- 11932174 TI - Antegrade palliation for diminutive pulmonary arteries in Tetralogy of Fallot. AB - OBJECTIVES: The purpose of this study was to evaluate the outcome following palliative reconstruction of right ventricular outflow tract in Tetralogy of Fallot (TOF) with diminutive pulmonary arteries with central and peripheral stenosis. METHODS: Between 1986 and 1999 in 15 children with the diagnosis of TOF palliative reconstruction of the right ventricular outflow tract without closure of the ventricular septal defect (VSD) was performed. All patients were not suitable for an AP-Shunt because of a diminutive pulmonary vascular bed. Six patients were younger than 1 year at operation. RESULTS: There was one hospital death (6.7%) in a child with additional aortic valve insufficiency in multi-organ failure. Although the postoperative course was prolonged (median duration on ICU: 8 days) and complicated by congestive heart failure, clinically the 14 patients discharged improved significantly. The arterial oxygen saturation increased from 67 to 93% (P<0.001), the hemoglobin decreased from 16.1 to 13.3g/l (P=0.02) and hematocrit from 0.52 to 0.40 (P=0.06). In control angiography, the McGoon Index increased in the average from 1.01 to 1.95 (P<0.001). VSD closure was performed in 12 patients (median: 2.5 years after initial operation) with one perioperative death. A homograft had to be implanted in seven patients and a mechanical prosthesis in the right ventricular outflow tract in one. One late death occurred due to ventricular arrhythmia 12 years after antegrade palliation (11 years after corrective operation). CONCLUSIONS: The antegrade palliation seems to be an adequate strategy for the treatment of selected children with diminutive pulmonary arteries in TOF, who were not candidates for primary correction or an AP-Shunt. PMID- 11932175 TI - Surgery for post infarction ventricular septal defect (VSD): risk factors for hospital death and long term results. AB - OBJECTIVE: Repair of post infarction ventricular septal defect (VSD) is still a challenging procedure with a high risk of recurrence of the VSD and subsequent mortality. The aim of this retrospective study was to assess if technical change in the surgical procedure was followed by an improvement in recurrence of the VSD and operative results. METHOD: This retrospective study from 1971 to 2001 included 85 patients operated on early (<15 days) after the occurrence of a post infarction VSD. Double patch technique was introduced in 1986. A total of 44 variables were studied by a uni- and multivariate analysis. RESULTS: Hospital death occurred in 36 patients. Significant factors for hospital mortality included: preoperative and evolution of the clinical status, right ventricular function and type of repair (one or two patches). Moreover, no recurrence was observed in patients repaired with the double patch technique (P=0.09). None of the studied variables were significant for long term survival. Concomitant CABG was not associated with higher hospital mortality and long-term survival rate was similar in patients with or without concomitant CABG. CONCLUSION: The use of the double patch technique and glue by avoiding recurrence of the VSD played a role in the reduction of the hospital mortality. This technique has to be recommended in the early repair of post infarction VSD. Concomitant CABG can be done safely to control the added risk of an associated coronary artery lesion. PMID- 11932176 TI - Short-term and 5-year outcome after primary isolated coronary artery bypass graft surgery: results of risk stratification in a bilocation center. AB - OBJECTIVE: We retrospectively investigated the short and mid-term outcome of non emergent primary isolated coronary artery bypass graft (CABG) surgery in relation to risk stratification in the fully equipped university location (FE) and the low volume, limited facility location (LVLF) of our department. METHODS: Between September 1995 and December 1996, 832 patients were referred to our department to undergo a primary isolated CABG operation. The surgical team selected 482 patients (58%) as being at low-risk. These were treated in the LVLF hospital. The other 350 patients with mixed-risk were treated in the FE hospital. The selection consisted primarily of exclusion of patients with moderate or poor left ventricular function, severe COPD or renal impairment, from surgery in the LVLF location. Finally, the prognostic value of the EuroSCORE and the Parsonnet score was tested on our patient population. RESULTS: Overall in-hospital mortality was 1.6% (13 patients). One patient died in the LVLF group (0.2%) and 12 patients (3.4%) in the FE group. LVLF patients experienced less complications during the hospital period compared to the FE patients (5 versus 21%; P=0.0001). The Parsonnet risk model and the EuroSCORE risk model showed both a good relation with in-hospital mortality. After discharge, an increased risk of late mortality was observed up to 1 year postoperative in the FE group compared to the LVLF group (2.7 versus 0.5%; P=0.01). Risk factors for 5-year mortality were pre operative renal impairment (blood creatinine >150 micromol/l) (hazard ratio (HR): 2.8; 95% confidence interval (CI): 1.4-5.5), diabetes (HR: 2.1; 95% CI: 1.3-3.5), impaired LVEF (HR: 1.9; 95% CI: 1.2-3.0), COPD (HR: 1.9; 95% CI: 1.1-3.5) and older age (HR: 1.07 per year; 95% CI: 1.01-1.10). Lipid-lowering therapy was a predictor of lower mortality at 5-years (HR: 0.5; 95% CI: 0.4-0.9). CONCLUSION: By careful decision making, selection of low-risk patients for a low volume and limited facility location resulted in excellent in-hospital survival with very low complication rates. PMID- 11932177 TI - Vascular complications of intra-aortic balloon insertion in patients undergoing coronary reavscularization: analysis of 911 cases. AB - OBJECTIVE: Intra-aortic balloon pump (IABP) is a well-accepted and widely used mechanical circulatory support in cardiac surgical practice. We evaluated the vascular complications of IABP and risk factors associated with the development of these complications in patients undergoing myocardial revascularization. METHODS: Between January 1994 and December 2000, a total of 911 patients undergoing coronary artery bypass grafting received IABP. The preoperative risk factors, balloon-related variables and vascular complications were studied and analyzed. Univariate and multivariate analyses were performed to identify risk factors for the development of vascular complications. RESULTS: Mean age of the patients was 59.2+/-9.1 years and 10.5% of the patients were female. The incidence of diabetes and peripheral vascular disease was 41.1 and 8.5%, respectively. The mean Parsonnet score was 11.8+/-4.6. IABP was inserted by percutaneous technique in 96.8% of patients. The duration of IABP therapy ranged from 20 h to 21 days (mean 3.8 days). Fifty-four (5.9%) patients developed major and 53 (5.8%) patients developed minor vascular complications. Ischaemia of the limb, requiring thromboembolectomy, developed in 25 (2.7%) patients. Patients who received IABP preoperatively had higher incidence of major vascular complications as compared to patients who received IABP in operating room before induction of anaesthesia. Multiple logistic regression analysis revealed age, triple vessel disease, indications of IABP therapy (unstable angina, cardiac arrhythmia and haemodynamic instability), left ventricular aneurysm surgery and use of balloon with sheath as independent risk factors for the development of vascular complications. CONCLUSIONS: IABP therapy is associated with certain vascular complications, which should always be kept in mind before insertion of a balloon. The use of a balloon without sheath and proper evaluation of peripheral circulation can help to minimize the development of vascular complications. PMID- 11932178 TI - Real-time and continuous monitoring of myocardial blood flow using a thermal diffusion method. AB - OBJECTIVES: There has been no method which can measure regional myocardial blood flow in real-time and continuously. The purpose of this study was to validate myocardial blood flow measurement using a thermal diffusion method. For this purpose, myocardial blood flow measurement was performed using the thermal diffusion method and the electrolytic hydrogen clearance method. METHODS: Seven pigs were used for this study, six were for comparison between the thermal diffusion and electrolytic hydrogen clearance methods, and one was for demonstration of myocardial blood flow measurement using the thermal diffusion method on a beating heart coronary artery bypass model with ischemic preconditioning. RESULTS: A good correlation was found between myocardial blood flow values obtained by the electrolytic hydrogen clearance method and 1/V values obtained by a thermal diffusion probe, the correlation coefficient was 0.841 (P<0.001). During the beating heart coronary artery bypass, the regional myocardial blood flow was recorded in real-time and continuously. CONCLUSIONS: This study demonstrated measurement of regional myocardial blood flow using the thermal diffusion method for the first time and simultaneous measurement using the electrolytic hydrogen clearance method for calibration. It provided a real time and continuous myocardial blood flow measurement and has a potential to contribute to progress in beating-heart surgery. PMID- 11932179 TI - The effect of chronic L-arginine administration on vascular recovery following cold cardioplegic arrest in rats. AB - OBJECTIVE: Acute administration of L-arginine (LA), the physiological substrate of nitric oxide, has been used as a strategy for myocardial protection during ischemia-reperfusion. The aim of this study was to assess the effects of chronic oral LA administration on vascular functions and morphology after prolonged cold cardioplegic arrest. METHODS: Adult male Sprague-Dawley rats (600-650 g) were divided into control and LA groups, which received LA (4 mg/ml) for 6 weeks. Two experimental protocols were carried out. (1) Isolated rat heart perfusion was performed and hearts were subjected to ischemia for 4 h at 4 degrees C using cold crystalloid cardioplegia (n=8 in LA, n=7 in control). Endothelial and vascular smooth muscle functions were assessed through observations of pre- and post ischemic coronary flow response to 5-hydroxytryptamine (5-HT) and glyceryl trinitrate (GTN) (%5-HT and %GTN, respectively). (2) Semi-quantitative assessment of tissue morphology was conducted after the same ischemia-reperfusion protocol (n=4 in each group). RESULTS: The LA group showed significantly better recovery (post-/pre-ischemic value) of %5-HT (97.0+/-65.6 versus 21.5+/-25.7%, P=0.015) and %GTN (124.5+/-117.6 versus 47.7+/-16.6%, P=0.021). The histological assessment showed no significant differences between the two groups. CONCLUSIONS: Chronic oral administration of LA significantly ameliorated the postischemic recovery of endothelial and vascular smooth muscle functions after cold cardioplegic arrest in rats. PMID- 11932180 TI - Images in cardio-thoracic surgery. Descending aortic aneurysm following ascending to descending bypass in the treatment of aortic re-coarctation. PMID- 11932181 TI - Use of an ultrasonic scalpel in the open-heart reoperation of a patient with pacemaker. AB - Electrocautery has a potential risk of serious pacemaker dysfunction in patients with implanted pacemaker. Here we present the safe and efficient use of ultrasonic scalpel (Harmonic scalpel) for the first time in a patient with implanted pacemaker undergoing open-heart reoperation. PMID- 11932182 TI - Tricuspid bioprosthesis replacement with cryopreserved mitral homograft. AB - We report the case of replacement of a dysfunctional tricuspid Hancock bioprosthesis by a cryopreserved mitral homograft. Tricuspid bioprosthesis was approached on a beating heart. The mitral homograft was orientated so as the anatomic anterior leaflets corresponding and a semi rigid prosthetic ring was inserted. At 1 year follow-up, the patient's clinical condition and echocardiographic results were satisfactory. PMID- 11932183 TI - Primary omental-flap surgery for post-sternotomy acute medistinitis. PMID- 11932184 TI - Technical refinements of omentopexy and pectoralis myoplasty for poststenotomy mediastinitis. PMID- 11932185 TI - Retrograde extension of type B dissection after endovascular stent graft repair. PMID- 11932188 TI - Changes in the current approach do not qualify prosthesis-patient mismatch either (reply to Dumesnil et al., EJCTS 2002;21:157-158). PMID- 11932189 TI - Introduction. Microbiological food safety. AB - This current focus covers a broad range of emerging microbiological issues in food safety, from chronic effects of campylobacterosis and bacterial antimicrobial resistance to microbial survival and growth on fresh fruits and vegetables, and advanced technologies for detection and inactivation of foodborne pathogens. PMID- 11932190 TI - Chronic effects of Campylobacter infection. AB - Campylobacter jejuni is one of the most common causes of bacterial gastroenteritis and chronic sequelae, such as reactive arthritis and Guillain Barre syndrome (GBS), are known to follow uncomplicated infections. While little is known about reactive arthritis following Campylobacter infection, our knowledge on the pathogenesis of Campylobacter-induced GBS is expanding rapidly and is summarized in this review. PMID- 11932191 TI - Antimicrobial resistance of foodborne pathogens. AB - Emergence of bacterial antimicrobial resistance has become a serious problem worldwide. While much of the resistance observed in human medicine is attributed to inappropriate use in humans, there is increasing evidence that antimicrobial use in animals selects for resistant foodborne pathogens that may be transmitted to humans as food contaminants. PMID- 11932192 TI - Ecological factors influencing survival and growth of human pathogens on raw fruits and vegetables. AB - Outbreaks of human infections associated with consumption of raw fruits and vegetables have occurred with increased frequency during the past decade. Factors contributing to this increase may include changes in agronomic and processing practices, an increase in per capita consumption of raw or minimally processed fruits and vegetables, increased international trade and distribution, and an increase in the number of immuno-compromised consumers. A general lack of efficacy of sanitizers in removing or killing pathogens on raw fruits and vegetables has been attributed, in part, to their inaccessibility to locations within structures and tissues that may harbor pathogens. Understanding the ecology of pathogens and naturally occurring microorganisms is essential before interventions for elimination or control of growth can be devised. PMID- 11932193 TI - Biosensor technologies for detecting microbiological foodborne hazards. AB - The convergence of molecular biology and miniaturized instrumentation has accelerated development of biosensors with the specifications necessary to support pathogen reduction and quality programs in the food supply. Advances in optoelectronics, thin layer deposition, and microfabrication have provided many options for achieving microbiological detection goals. Some promising technologies are reviewed. PMID- 11932194 TI - Alternative food-preservation technologies: efficacy and mechanisms. AB - High-pressure processing, ionizing radiation, pulsed electric field and ultraviolet radiation are emerging preservation technologies designed to produce safe food, while maintaining its nutritional and sensory qualities. A sigmoid inactivation pattern is observed in most kinetic studies. Damage to cell membranes, enzymes or DNA is the most commonly cited cause of death of microorganisms by alternative preservation technologies. PMID- 11932195 TI - CD8(+) T-cell homeostasis after infection: setting the 'curve'. AB - Antigen (Ag)-specific CD8(+) T-cell responses exhibit remarkably similar kinetics after different types of infection. Starting from levels that are virtually undetectable in vivo, pathogen-specific naive CD8(+) T cells are precisely regulated to go through rapid expansion and contraction (death) phases, achieving memory levels of Ag-specific CD8(+) T cells that are maintained for the life of the host. However, the exact mechanisms used to achieve appropriate and reproducible CD8(+) T-cell homeostasis in response to diverse pathogens remain to be determined. The possibility that early events after infection regulate major features of Ag-specific CD8(+) T-cell homeostasis will be discussed here. PMID- 11932196 TI - Macrophage migration inhibitory factor (MIF): mechanisms of action and role in disease. AB - Macrophage migration inhibitory factor (MIF) is a unique cytokine and critical mediator of host defenses with a role in septic shock and chronic inflammatory and autoimmune diseases. Its mechanism of action is incompletely understood. Here, we attempt to correlate current knowledge on the molecular pathways of MIF activity with its functions in immunity and disease. PMID- 11932197 TI - Similar and divergent features in mammalian and yeast prions. AB - Mammalian transmissible spongiform encephalopathies are likely due to the propagation of an abnormal form of a constitutive protein instead of traditional genetic material (nucleic acids). Such infectious proteins, which are termed prions, exist in yeast. They are at the origin of a number of phenotypes that are inherited in a non-Mendelian manner. These prions are very useful to dissect the molecular events at the origin of this structure-based inheritance. The properties of mammalian and yeast prions are presented and compared. This review highlights a number of similarities and differences. PMID- 11932198 TI - Control of Mycobacterium bovis infections and the risk to human populations. AB - Conventional control methods based on test-and-slaughter policies have, in several countries, led to the successful eradication of bovine tuberculosis in cattle. However, new approaches for control of bovine tuberculosis are required in developing countries and those with a wildlife reservoir of infection. Recent developments include improved diagnostics and evaluation of new vaccination strategies. PMID- 11932199 TI - Staphylococcus epidermidis infections. AB - The opportunistic human pathogen Staphylococcus epidermidis has become the most important cause of nosocomial infections in recent years. Its pathogenicity is mainly due to the ability to form biofilms on indwelling medical devices. In a biofilm, S. epidermidis is protected against attacks from the immune system and against antibiotic treatment, making S. epidermidis infections difficult to eradicate. PMID- 11932200 TI - Molecular and cellular biology of Borna disease virus infection. AB - Borna disease virus (BDV) is a noncytolytic, neurotropic RNA virus that causes neurobehavioral disorders in a wide variety of warm-blooded animals. Recent evidence has revealed that BDV uses a unique strategy in its transcription and replication and directly affects cellular functions of infected central nervous systems. BDV research will provide new insights not only into the biology of neurotropic RNA virus but also into neuropsychiatry. PMID- 11932201 TI - The molecular basis of Streptococcus equi infection and disease. AB - Streptococcus equi is the aetiological agent of strangles, one of the most prevalent diseases of the horse. The animal suffering and economic burden associated with this disease necessitate effective treatment. Current antibiotic therapy is often ineffective and thus recent attention has focused on vaccine development. A systematic understanding of S. equi virulence, leading to the identification of targets to which protective immunity can be directed, is a prerequisite of the development of such a vaccine. Here, the virulence factors of S. equi are reviewed. PMID- 11932202 TI - Genetic immunization of neonates. AB - The vaccination of neonates is generally difficult due to immaturity of the immune system, higher susceptibility to tolerance and potential negative interference of maternal antibodies. Studies carried out in rodents and non-human primates showed that plasmid vaccines expressing microbial antigens, rather than inducing tolerance, triggered significant humoral and cellular immunity with a Th1 component. The ability of bacterial CpG motifs to activate immature antigen presenting cells is critical for the neonatal immunogenicity of DNA vaccines. In addition, the endogenous production of antigen subsequent to transfection of antigen-presenting cells may explain the lack of inhibition by maternal antibodies of cellular responses. Together, these features make the plasmid vaccines an appealing strategy to prime immune responses against foreign pathogens, during early life. In combination with subsequent boosting using conventional vaccines, DNA vaccine-based regimens may provide a qualitatively superior immunity against microbes. Thorough understanding of immunomodulatory properties of plasmid-vectors may extend their use for early prophylaxis of inflammatory disorders. PMID- 11932203 TI - Immune cognition and vaccine strategy: beyond genomics. AB - I.R. Cohen's work on immune cognition has profound implications for vaccine strategies when simple elicitation of sterilizing immunity fails, given Nisbett's analysis showing that cognition by the central nervous system is culturally determined. We reinterpret West African cultural variation in immune response to malaria, and the US cultural variation in HIV transmission, from this perspective, which does not reify 'race'. PMID- 11932204 TI - Negative regulation of nuclear factor-kappaB activation and function by glucocorticoids. AB - Glucocorticoids (GCs) exert their anti-inflammatory and antiproliferative effects principally by inhibiting the expression of cytokines and adhesion molecules. Mechanistically, GCs diffuse through the cell membrane, and bind to their inactive cytosolic receptors (GRs), which then undergo conformational modifications that allow for their nuclear translocation. In the nucleus, activated GRs modulate transcriptional events by directly associating with DNA elements, compatible with the GCs response elements (GRE) motif, and located in variable copy numbers and at variable distances from the TATA box, in the promoter region of GC-responsive genes. In addition, activated GRs also acted by antagonizing the activity of transcription factors, in particular nuclear factor kappaB (NF-kappaB), by direct and indirect mechanisms. GCs induced gene transcription and protein synthesis of the NF-kappaB inhibitor, IkappaB. Activated GR also antagonized NF-kappaB activity through protein-protein interaction involving direct complexing with, and inhibition of, NF-kappaB binding to DNA (Simple Model), or association with NF-kappaB bound to the kappaB DNA site (Composite Model). In addition, and according to the Transmodulation Model, GRE-bound GR may interact with and inhibit the activity of kappaB-bound NF kappaB via a mechanism involving cross-talk between the two transcription factors. Lastly, GR may compete with NF-kappaB for nuclear coactivators, including CREB binding protein and p300, thereby reducing and inhibiting transcriptional activation by NF-kappaB. It should be noted that, in exerting its effect, activated GR did not affect the correct assembly of the pre-initiation (DAB) complex, but acted rather more proximally in inhibiting the correct assembly of transcription factors in the promoter region, and thus transcriptional initiation. PMID- 11932205 TI - Prolonged dietary treatment with conjugated linoleic acid stimulates porcine muscle peroxisome proliferator activated receptor gamma and glutamine-fructose aminotransferase gene expression in vivo. AB - Peroxisome proliferator activated receptors (PPARs) represent a family of DNA binding proteins that are activated by a variety of dietary and endogenous fatty acids. The PPAR proteins are expressed throughout the body and are the target of a variety of lipidaemic and insulin sensitizing drugs. Conjugated linoleic acid (CLA) is a collective name for octadecadienoic acid isomers with conjugated double bonds, which can also act as ligands for some of the PPAR family. To gain better understanding of the long-term effects of PPAR activation, CLA was fed at 11 g/kg of feed for 45 days to castrated male pigs (barrows). These barrows had a significant repartitioning of subcutaneous fat to lean tissue in the carcass: fat was reduced by 9 x 2% and lean muscle was increased by 3 x 5%, but intramuscular fat content was also increased by 14% (P<0 x 05). PPARgamma, glutamine-fructose aminotransferase (GFAT), adipocyte fatty acid binding protein (AFABP), but not PPARalpha mRNA levels were significantly increased (P<0 x 05) in the CLA-fed pigs. The increased expression of PPARgamma and AFABP indicates that CLA induced the development of preadipocytes from stromal-vascular (s-v) stem cells to promote intramuscular fat content. The increase in the expression of GFAT mRNA indicates that the glucose supply of the muscle cells had been increased with the CLA diet, possibly sparing intramuscular fatty acid reserves. PMID- 11932206 TI - Changes in ontogenetic expression of estrogen receptor alpha and not of estrogen receptor beta in the female rat reproductive tract. AB - To evaluate ontogenetic expression and localization of estrogen receptor (ER) alpha and beta in fetal female rat reproductive tract, competitive RT-PCR and immunohistochemistry were performed. Expression levels for Mullerian ERalpha, ERbeta1 and ERbeta2 mRNAs were determined by competitive RT-PCR. ERalpha expression on gestational day (GD) 15 x 5 increased 4 x 4-fold by GD 21 x 5, whereas both ERbeta1 and ERbeta2 gene expression were maintained at lower constant levels compared with ERalpha during development. ER immunolocalization was evaluated within three regions along the Mullerian duct axis; these were proximal, middle and caudal, which differentiate into oviduct, uterus and upper vagina respectively. Nuclear ERalpha was localized predominantly in proximal Mullerian epithelium, and middle and caudal Mullerian mesenchyme on GDs 15 x 5-21 x 5. Staining intensity for ERalpha increased with development in all regions. However, ERbeta immunoreactivity was not detected in any region during prenatal life after separate staining with three different polyclonal anti-rat ERbeta antibodies. These findings provide fundamental information critical for clarifying the species-specific physiological roles of ER subtypes during fetal development and for investigating the tissue-specific mechanisms underlying the prenatal response to estrogen and estrogen receptor agonists. PMID- 11932207 TI - Hepatocyte growth factor stimulates proliferation of pancreatic beta-cells particularly in the presence of subphysiological glucose concentrations. AB - We investigated the role of hepatocyte growth factor (HGF) in beta-cell growth and its complex intracellular signal transduction pathways. Cell proliferation was measured in the beta-cell line INS-1 using [3H]thymidine incorporation. Activation of mitogenic signaling proteins was assessed using co immunoprecipitation, immunoblot analysis and specific protein activity inhibitors in proliferation assays. HGF (1 x 375 nM) increased INS-1 cell proliferation in the presence of 3-24 mM glucose up to 45-fold vs unstimulated controls. HGF exceeded the effect of glucose alone (2 x 2-fold at 3 mM glucose and 1 x 7-fold in the presence of 15 mM glucose). The HGF-induced INS-1 cell proliferation was further increased by addition of IGF-I or GH. Stimulation with HGF activated the JAK-2/STAT-5 pathway with a subsequent activation of phosphatidylinositol-3' kinase (PI3'K). PI3'K activation was necessary for HGF- and glucose-stimulated INS-1 cell proliferation. The effect of PI3'K was mediated via 70 kDa S6 kinase and protein kinase B, which showed maximum activation in the presence of 3-6 mM glucose. Protein kinase C was essential for HGF-induced INS-1 cell proliferation. The HGF effect was also mediated at low glucose concentrations via insulin receptor substrate 4 (IRS-4) whereas other IRS proteins did not show any activation. High glucose concentrations also showed an increased IRS-4/PI3'K binding and therefore activation. In conclusion, beta-cell proliferation is mediated via complex interacting signal transduction pathways. HGF, in contrast to other growth factors, seems to be of importance particularly in the presence of low glucose concentrations and therefore takes a special role in this complex concert. PMID- 11932208 TI - Calcitonin: characterisation and expression in a teleost fish, Fugu rubripes. AB - The present report describes the structure and expression of the calcitonin gene in Fugu rubripes. It is composed of 4 exons and 3 introns. Splicing of exons 1, 2 and 3 generates the calcitonin pre-proprotein, while splicing of exons 1, 2 and 4 generates calcitonin gene-related protein (CGRP). Exons 1 and 2 encoding the signal sequence and the N-terminal peptide are common in both the gene products and this gene organisation has been conserved in human, rat, chicken and salmon. The gene environment around calcitonin in Fugu has been poorly conserved when compared with human, apart from a small gene cluster. The calcitonin gene in Fugu has a widespread tissue distribution but it is most highly expressed in the brain. The abundance of gene expression in the ultimobranchial gland and the pituitary indicates that these are important sites of production and that the peptide is probably secreted into the circulation and/or acts as a paracrine or autocrine controlling factor. Whilst the function of calcitonin in fish is still largely unknown, the distribution described here suggests that one of the potential functions may be as a neuropeptide. PMID- 11932209 TI - Differential regulation of aldosterone synthase and 11beta-hydroxylase transcription by steroidogenic factor-1. AB - 11beta-Hydroxylase (hCYP11B1) and aldosterone synthase (hCYP11B2) are closely related isozymes with distinct roles in cortisol and aldosterone production respectively. Aldosterone synthase catalyzes the final step in aldosterone biosynthesis and is expressed only in the zona glomerulosa of the normal adrenal. 11beta-Hydroxylase catalyzes the final reaction in the production of cortisol and is expressed at higher levels in the zona fasciculata. The mechanisms causing differential expression of these genes are not well defined. Herein, we demonstrate contrasting roles for the orphan receptor steroidogenic factor-1 (SF 1) in the regulation of human (h) CYP11B1 and hCYP11B2. Human NCI-H295R (H295R) or mouse Y-1 cells were transiently transfected with luciferase reporter constructs containing 5'-flanking regions of hCYP11B1, hCYP11B2, human 17alpha hydroxylase (hCYP17), human cholesterol side-chain cleavage (hCYP11A1) or mouse (m) cyp11b2 (mcyp11b2). Co-transfection of vectors encoding SF-1 increased expression of hCYP11B1, hCYP11A1 and hCYP17 constructs, but inhibited hCYP11B2 reporter activity. Murine, bovine and human SF-1 were unable to increase transcription of hCYP11B2 in H295R cells. Both hCYP11B2 and mcyp11b2 promoter constructs were inhibited similarly by human SF-1. In mouse Y-1 cells, reporter expression of hCYP11B2 and mcyp11b2 was very low compared with hCYP11B1 constructs, suggesting that this adrenal cell model may not be appropriate for studies of CYP11B2. Electrophoretic mobility shift assay demonstrated that SF-1 interacted with an element from both hCYP11B1 and hCYP11B2. However, mutation of this element, termed Ad4, did not prevent agonist stimulation of hCYP11B2 by angiotensin II or forskolin but blocked activity of hCYP11B1. In some, but not all, reports of genetic linkage analysis, a naturally occurring single nucleotide polymorphism within the Ad4 element of hCYP11B2 (-344C/T) has been associated with cardiovascular disease. Herein, we have demonstrated that this polymorphism influenced binding of SF-1 in electrophoretic mobility shift assays, with the C allele binding SF-1 more strongly than the T allele. However, when hCYP11B2 constructs containing these alleles were transfected into H295R cells, there was no difference in agonist-stimulated expression or the response of either reporter construct to co-expression with human SF-1. Taken together, these data suggest that SF-1 and the Ad4 element are not major regulators of adrenal hCYP11B2 gene expression. Thus far, hCYP11B2 is the first steroid hydroxylase gene which is not positively regulated by SF-1. PMID- 11932210 TI - Expression and dimerization of the rat activin subunits betaC and betaE: evidence for the ormation of novel activin dimers. AB - Activins are cytokines of the transforming growth factor beta family, which plays a central role in the determination of cell fate and the regulation of tissue balance. Family members are composed of two subunits and this dimerization is critical for liganding their cognate receptors and execution of proper functions. In the current study we focused on the localization of activin betaA, betaB, betaC and betaE subunits in the adult rat and analyzed the composition of putative activin beta dimers. By dissecting tissue distribution of various activins, we found that the liver, in particular the hepatocytes, is the major source for activin betaC and betaE transcripts, since other tissues almost failed to express these isoforms. In sharp contrast, the emergence of activin betaA and betaB appeared ubiquitous. Using a highly selective proteome approach, we were able to identify homo- as well as heterodimers of individual activin subunits, indicating a high redundancy of ligand composition. Certainly, this broad potential to homo- and heterodimerize has to be considered in future studies on activin function. PMID- 11932211 TI - Legal issues in teleradiology-distant thoughts! AB - Advancements in computer technology and telecommunications have meant that all diagnostic images can now be acquired as digital signals, however the ethicolegal concepts surrounding this innovation remain unclear. In the UK there are limited practice guidelines on legal issues relating specifically to telemedicine or teleradiology. It is not yet clear whether the current law relates to telemedicine in the same way that it does for other medical specialties, or whether telemedicine raises new legal issues that need clarification. This article attempts to outline some of the potential legal issues, but the absence of case law and legislation in this area will ensure that many questions remain unanswered. The legal implications of teleradiology/telemedicine are addressed, and literature, laws and professional guidelines from the UK, USA, Australia and New Zealand are reviewed, focusing on the American licensure laws, professional relationships with patients with regards to liability, responsibility, accountability and duty of care issues, as well as issues of missed diagnosis, misdiagnosis, security and confidentiality. Teleradiology, while being actively practised worldwide, gives rise to many unanswered medicolegal questions. It is suggested that guidelines need to be implemented to safeguard patients and professionals alike. PMID- 11932212 TI - Acute vertebral body compression fractures: discrimination between benign and malignant causes using apparent diffusion coefficients. AB - Diffusion weighted MRI was performed on patients with acute vertebral body compression. The usefulness of the apparent diffusion coefficient (ADC) in differentiating between benign and malignant fractures was evaluated. A total of 49 acute vertebral body compression fractures were found in 32 patients. 25 fractures in 18 patients were due to osteoporosis, 18 fractures in 12 patients were histologically proven to be due to malignancy, and 6 fractures in 2 patients were due to tuberculosis. Signal intensities on T(1) weighted, short tau inversion recovery (STIR) and diffusion weighted images were compared. ADC values of normal and abnormal vertebral bodies were calculated. Except for two patients with sclerotic metastases, benign acute vertebral fractures were hypointense and malignant acute vertebral fractures were hyperintense with respect to normal bone marrow on diffusion weighted images. Mean combined ADCs (ADC(cmb); average of the combined ADCs in the x, y and z diffusion directions) were 0.23 x 10(-3) mm(2) s( 1) in normal vertebrae, 0.82 x 10(-3) mm(2) s(-1) in malignant acute vertebral fractures and 1.94 x 10(-3) mm(2) s(-1) in benign acute vertebral fractures. The differences between ADC(cmb) values were statistically significant (p<0.001). The ADC is useful in differentiating benign from malignant acute vertebral body compression fractures, but there may be overlapping ADC values between malignant fractures and tuberculous spondylitis. PMID- 11932213 TI - Assessment of fetal anatomy in the first trimester using two- and three dimensional ultrasound. AB - The objective of this study was to perform a complete anatomical survey of the fetus at 12-13 weeks gestation using stored volumes acquired by a three dimensional (3D) scanner. 159 consecutive women at 12-13 weeks gestation who had a routine early pregnancy scan in our unit were recruited. A complete survey of the fetal anatomy was attempted by two-dimensional (2D) transabdominal and, if needed, transvaginal ultrasound. Then, using a 3D transvaginal probe, two volumes of the whole fetus were acquired. A complete anatomical survey (excluding anatomy of the heart) was attempted using the stored data. A complete anatomical survey was achieved in 93.7% (149) of cases with 2D ultrasound compared to 80.5% (128) of cases with 3D volume acquisition (p<0.001). The nuchal translucency was measured with 2D scanning in 98.7% of cases and in 91.8% of cases using 3D volumes. The mean time to perform a 2D scan was 12.2 min standard deviation (SD 3.4 min) while the mean time to obtain and examine the stored volumes was 8.4 min (SD 1.45 min, p<0.001). Real-time 2D ultrasound is still the best way to examine fetal anatomy in the first trimester. However, 3D ultrasound can be a useful addition to clinical practice, providing views not easily obtained by conventional 2D ultrasound. It can potentially minimize actual scanning time and provides an excellent way to store scanned data. PMID- 11932215 TI - The role of MRI in the evaluation of hip joint disease in clinical subtypes of juvenile idiopathic arthritis. AB - The purpose of this study was to evaluate the role of MRI in the assessment of hip joint involvement in clinical subtypes of juvenile idiopathic arthritis (JIA). 28 patients (mean age 12.5 years) with JIA (oligoarthritis 8, polyarthritis 13, systemic arthritis 7) were examined with T(2) weighted turbo spin echo and T(1) weighted spin echo (plain and contrast enhanced) sequences. The severity of joint involvement was evaluated using an MR grading score: grade 1=no contrast enhancement; grade 2=focal synovial contrast enhancement; grade 3=diffuse synovial contrast enhancement; grade 4=grade 3+diffuse synovial thickening; grade 5=grade 4+villonodular synovial thickening; and grade 6=grade 5+cartilage and subchondral bone erosions. MRI was abnormal in 57.1% of cases (25% of oligoarthritis, 53.8% of polyarthritis and 100% of systemic arthritis). Clinical examination was positive in 32.1% of cases and was associated with higher MR grades (mean 4.6, SD 1.34) compared with a negative clinical examination, which was associated with lower MR grades (mean 1.78, SD 1.13) (p<0.001). Patients with active disease (mean grade 3.9, SD 2) had higher MR grades than those with inactive disease (mean grade 2.1, SD 1.4) (p<0.01). The MR grades were different in the three clinical subtypes: oligoarticular (mean 1.5, SD 1.06); polyarticular (mean 2.38, SD 1.55); and systemic (mean 4.85, SD 1.21) (F:12.3, p<0.001), with a significant difference between systemic arthritis and oligoarthritis, and between systemic arthritis and polyarthritis (p<0.001). MRI of the hip might be considered for inclusion in the study protocol of patients with JIA since it reveals joint involvement at early stages and provides a detailed evaluation of the extent of joint disease. PMID- 11932214 TI - Dynamic contrast enhanced MRI of the axilla in women with breast cancer: comparison with pathology of excised nodes. AB - Axillary lymph node status is the most important prognostic factor in breast cancer patients and is currently determined by surgical dissection. This study was performed to assess whether dynamic gadopentetate dimeglumine (Gd) enhanced MRI is an accurate method for non-invasive staging of the axilla. 47 women with a new primary breast cancer underwent pre-operative dynamic Gd enhanced MRI of the ipsilateral axilla. Lymph node enhancement was quantitatively analysed using a region of interest method. Enhancement indices and nodal area were compared with histopathology of excised nodes using a receiver operating characteristic (ROC) curve approach. 10 patients had axillary metastases pathologically and all had > or =1 lymph node with an enhancement index of >21% and a nodal area of >0.4 cm(2). 37 patients had negative axillary nodes pathologically. 20 of these had enhancement indices <21% and nodal areas <0.4 cm(2). Using this method, a sensitivity of 100%, a specificity of 56%, a positive predictive value of 38% and a negative predictive value of 100% could be achieved. Using this method of quantitative assessment, dynamic Gd enhanced MRI may be a reliable method of predicting absence of axillary nodal metastases in women with breast cancer, thereby avoiding axillary surgery in women with a negative MRI study. PMID- 11932216 TI - The influence of applicator angle on dosimetry in vaginal vault brachytherapy. AB - In vaginal vault brachytherapy, the critical normal tissues are bladder and rectum; doses to these tissues may be affected by the position of a single line applicator placed in the vagina. Dosimetry with the applicator lying at its "natural" angle in the vagina with the patient in the lithotomy position has been compared with the applicator held horizontal as defined by a spirit level in 30 consecutive patients. A mean change in angle of 19.7 degrees was found. This resulted in a mean decrease in ICRU (International Commission of Radiation Units and Measurements) rectal point dose when the applicator is horizontal of 12.9%, equivalent to a mean absolute dose reduction of 1.3 Gy for a prescription dose of 5.5 Gy at 5 mm depth. An increase in mean dose to the ICRU bladder point when the applicator is horizontal of 13.3%, equivalent to an absolute mean dose increase of 0.5 Gy per fraction for the same prescription dose, was also found. On the basis of these findings, it is recommended that vaginal vault brachytherapy is performed with a single line source held in the "corrected" horizontal position to reduce bowel dose as this is the most sensitive critical normal tissue. PMID- 11932218 TI - Irish X-ray departments demonstrate varying levels of adherence to European guidelines on good radiographic technique. AB - The Commission of European Communities (CEC) publication "European Guidelines on Quality Criteria for Diagnostic Radiographic Images" includes examples of good radiographic technique for a number of common X-ray examinations. If these guidelines are followed, compliance with dose and image quality criteria as specified in the CEC document should be demonstrated. Studies in England, Germany and Greece have shown that a number of X-ray departments are not using optimum techniques. The aim of this study is to demonstrate the level of adherence to CEC guidelines in Irish hospitals 3-4 years following publication of the above document. 16 hospitals were randomly chosen and the following details on technique and equipment were recorded for chest, abdomen, pelvis and lumbar spine examinations of standard sized patients: tube potential, focus-to-film distance, automatic exposure control (AEC), film-screen combination, X-ray tube filtration and secondary radiation grid. Varying levels of adherence to the guidelines were evident depending on the parameter being investigated, with no hospital demonstrating 100% compliance and no hospital demonstrating 100% non-compliance. For all parameters, with the exception of AEC use, the majority of hospitals exhibited non-adherence for at least one projection. The results suggest that if hospitals in Ireland observe the straightforward examples of good radiographic technique described in the CEC publication, significant reductions in collective dose can be achieved. PMID- 11932217 TI - Severe complications after hypofractionated high dose rate intracavitary brachytherapy following external beam irradiation for oesophageal carcinoma. AB - The purpose of this study was to evaluate severe complications that developed after high dose rate (HDR) intracavitary brachytherapy for oesophageal carcinoma. Six consecutive patients with oesophageal carcinoma were treated by external beam irradiation (60 Gy in 30 fractions over 6 weeks) followed by hypofractionated intracavitary HDR brachytherapy (10 Gy in 2 fractions). Two of the six patients were alive and well for more than 2-3 years following therapy, but three of the six patients developed treatment-related oesophageal fistulae and died. HDR intracavitary brachytherapy following external beam irradiation is an effective method for radical treatment of oesophageal carcinoma. However, hypofractionated HDR brachytherapy should be used with care. PMID- 11932219 TI - Paediatric personnel extremity dose study. AB - Concern has been expressed in paediatric radiology regarding the magnitude of the extremity dose received by attending personnel during routine fluoroscopic procedures and CT. Common procedures that may be of short duration in adults can be quite the opposite in paediatric patients. The extremities of attending personnel are more likely to be exposed to the primary beam and for a longer period of time owing to a variety of reasons such as assisting in the procedure or physically restraining the patient during the examination. During the period mid 1998 to mid 2000, two paediatric radiologists, four senior radiographers and two paediatric nurses were monitored using ring thermoluminescent dosemeters (TLDs). Each participant wore the ring TLD on either the left or right ring finger, depending on which hand the individual favoured. Left/right asymmetrical studies were not conducted, nor were records kept of whether an examination used a grid or gridless technique. Initial apprehension about higher paediatric fluoroscopic and CT extremity doses was dispelled as a result of this quantitative dosimetric study. PMID- 11932220 TI - X-ray dose and associated risks from radiofrequency catheter ablation procedures. AB - The objectives of this study were to quantify the ionizing radiation exposure to patient and operator during radiofrequency (RF) catheter ablation and to estimate the risks associated with this exposure. The study consisted of 50 RF ablation procedures, all performed in the same electrophysiology laboratory. Occupational dose to two cardiologists who performed the procedures was measured using film badges and extremity thermoluminescent dosemeters (TLDs). Absorbed dose to the patients' skin was measured using TLDs. Dose-area product (DAP) was also measured. The effective dose to the cardiologists was less than 0.15 mSv per month. The mean equivalent dose to the cardiologists' left hand and forehead was 0.24 mSv and 0.05 mSv, respectively, per RF ablation procedure, which was more than twice the mean dose for the other cardiology procedures carried out in the centre. Yearly occupational dose to the cardiologists was much lower than the relevant statutory dose limits. The mean skin dose, fluoroscopy time and DAP to patients were 0.81 Gy, 67 min and 123 Gycm(2), respectively, with a maximum of 3.2 Gy, 164 minutes and 430 Gycm(2), respectively. Mean effective dose to patients was 17 mSv, from which the excess risk of developing fatal cancer is 0.1%. Six of the patients (12%) received a skin dose above the threshold dose for radiation skin injury (2 Gy), but no skin injuries were reported. Patient skin dose and DAP were closely correlated and this allows DAP to be used to monitor patient skin dose in real-time. DAP levels were locally adopted as diagnostic reference levels (DRLs) that provide an indication during a procedure that a patient is at risk of suffering deterministic skin injury. PMID- 11932221 TI - Assessment of distortion in a three-dimensional rotational angiography system. AB - The purpose of this project was to determine the degree of geometrical distortion in a three-dimensional (3D) image volume generated by a digital fluorography system with rotational image acquisition capabilities. 3D imaging is a valuable adjunct in neuroangiography for visualization and measurement of cerebral aneurysms and for determination of the optimum projection for intervention. To enable spatially accurate 3D reconstruction the system must correct for geometrical distortion in the image intensifier television system as well as for deviations in gantry motion. 3D volumes were reconstructed from 100 X-ray projections acquired over a 180 degrees arc over a period of 8 s. A phantom was constructed to assess geometrical distortion in the three dimensions. The phantom consisted of 1 mm diameter ball bearings embedded in Perspex in a cubic lattice configuration. The ball bearings were placed at 20 mm intervals over a 140 mm cubic volume. Distortion was assessed by taking measurements between points of known separation and using a differential distortion measurement. The maximum error in the 3D location of objects was found to be 1.4 mm, while the differential distortion was found to range from -1.0% to +2.3%. The 3D images were found to have negligible visual distortion, enabling subjective assessments to be made with confidence to aid intervention. PMID- 11932222 TI - Intrabiliary rupture of hydatid cyst: diagnosis with MRI and hepatobiliary isotope study. AB - Intrabiliary rupture is the most common complication of hepatic hydatid cyst yet it is unusual, occurring in only 3-17% of cases. The diagnosis is rarely difficult on ultrasound and CT when typical radiological features are present. In rare cases of complete evacuation, when characteristic findings of hydatid cyst are absent or when there is no evidence of the previous existence of liver hydatid cyst, the diagnosis may be difficult. In difficult cases, MRI, MRCP, ERCP and (99)Tc(m)-mebrofenin hepatobiliary scintigraphy are employed. We present a rare case of surgical obstructive jaundice due to rupture of a liver hydatid cyst into the biliary tract and gall bladder, with complete evacuation of its contents leading to misdiagnosis on CT and ultrasound. MRCP and (99)Tc(m)-mebrofenin hepatobiliary scintigraphy were able to establish a firm pre-operative diagnosis. PMID- 11932223 TI - Cryptococcus infection in a patient with nasopharyngeal carcinoma: imaging findings mimicking pulmonary metastases. AB - An asymptomatic pulmonary mass was found in a 42-year-old unmarried male with controlled nasopharyngeal carcinoma (NPC) during routine follow-up chest radiography 8 months following completion of radiotherapy. Chest CT demonstrated a 3x2 cm(2) left lower lobe (LLL) mass, with further small nodules in the same lobe. A presumptive diagnosis of lung metastases was made, and the patient underwent surgical resection with left lower lobectomy and mediastinal lymph node dissection. Pathologic examination of the masses in the LLL revealed granulomatous inflammation with cryptococcus infection. The dissected lymph nodes revealed anthracosis. The patient received 6 months of antifungal treatment with fluconazole. His NPC showed no evidence of local recurrence or distant metastases. Recognition that pulmonary cryptococcus infection can mimic metastases is important in reaching the correct diagnosis and therefore determining the correct treatment. PMID- 11932224 TI - Subcutaneous calcification following chest wall and breast irradiation: a late complication. AB - Subcutaneous calcification as a complication of chest wall irradiation has only been described once before in the literature. Six patients who developed heavy calcification of soft tissue following chest wall and breast irradiation are described here, and relevant literature is reviewed. PMID- 11932225 TI - Scrotal calcification: ultrasound appearances, distribution and aetiology. AB - This pictorial review illustrates the ultrasound appearances of scrotal calcification, distinguishing between intratesticular and extratesticular calcification. Intratesticular calcification may be due to phleboliths, spermatic granulomas or vascular calcification, or it may occur in association with tumours. Extratesticular calcification is more frequently encountered and is usually related to previous inflammatory disease of the epididymis. Testicular microlithiasis, a rare condition characterized by multiple scattered echogenic foci within the testis, is produced by the formation of microliths from degenerating cells in the seminiferous tubules. Testicular microlithiasis has been demonstrated as an incidental finding as well as in association with both benign and malignant tumours of the testis. PMID- 11932226 TI - A nipper from Nippon? PMID- 11932227 TI - Guidelines: commentary and controversy. PMID- 11932228 TI - Bacterial adhesion: seen any good biofilms lately? AB - The process of surface adhesion and biofilm development is a survival strategy employed by virtually all bacteria and refined over millions of years. This process is designed to anchor microorganisms in a nutritionally advantageous environment and to permit their escape to greener pastures when essential growth factors have been exhausted. Bacterial attachment to a surface can be divided into several distinct phases, including primary and reversible adhesion, secondary and irreversible adhesion, and biofilm formation. Each of these phases is ultimately controlled by the expression of one or more gene products. Ultrastructurally, the mature bacterial biofilm resembles an underwater coral reef containing pyramidal or mushroom-shaped microcolonies of organisms embedded within an extracellular glycocalyx, with channels and cavities to allow the exchange of nutrients and waste. The biofilm protects its inhabitants from predators, dehydration, biocides, and other environmental extremes while regulating population growth and diversity through primitive cell signals. From a physiological standpoint, surface-bound bacteria behave quite differently from their planktonic counterparts. Recognizing that bacteria naturally occur as surface-bound and often polymicrobic communities, the practice of performing antimicrobial susceptibility tests using pure cultures and in a planktonic growth mode should be questioned. That this model does not reflect conditions found in nature might help explain the difficulties encountered in the management and treatment of biomedical implant infections. PMID- 11932229 TI - Biofilms: survival mechanisms of clinically relevant microorganisms. AB - Though biofilms were first described by Antonie van Leeuwenhoek, the theory describing the biofilm process was not developed until 1978. We now understand that biofilms are universal, occurring in aquatic and industrial water systems as well as a large number of environments and medical devices relevant for public health. Using tools such as the scanning electron microscope and, more recently, the confocal laser scanning microscope, biofilm researchers now understand that biofilms are not unstructured, homogeneous deposits of cells and accumulated slime, but complex communities of surface-associated cells enclosed in a polymer matrix containing open water channels. Further studies have shown that the biofilm phenotype can be described in terms of the genes expressed by biofilm associated cells. Microorganisms growing in a biofilm are highly resistant to antimicrobial agents by one or more mechanisms. Biofilm-associated microorganisms have been shown to be associated with several human diseases, such as native valve endocarditis and cystic fibrosis, and to colonize a wide variety of medical devices. Though epidemiologic evidence points to biofilms as a source of several infectious diseases, the exact mechanisms by which biofilm-associated microorganisms elicit disease are poorly understood. Detachment of cells or cell aggregates, production of endotoxin, increased resistance to the host immune system, and provision of a niche for the generation of resistant organisms are all biofilm processes which could initiate the disease process. Effective strategies to prevent or control biofilms on medical devices must take into consideration the unique and tenacious nature of biofilms. Current intervention strategies are designed to prevent initial device colonization, minimize microbial cell attachment to the device, penetrate the biofilm matrix and kill the associated cells, or remove the device from the patient. In the future, treatments may be based on inhibition of genes involved in cell attachment and biofilm formation. PMID- 11932231 TI - Dracunculiasis (Guinea worm disease) and the eradication initiative. AB - Dracunculiasis, also known as guinea worm disease, is caused by the large female of the nematode Dracunculus medinensis, which emerges painfully and slowly from the skin, usually on the lower limbs. The disease can infect animals, and sustainable animal cycles occur in North America and Central Asia but do not act as reservoirs of human infection. The disease is endemic across the Sahel belt of Africa from Mauritania to Ethiopia, having been eliminated from Asia and some African countries. It has a significant socioeconomic impact because of the temporary disability that it causes. Dracunculiasis is exclusively caught from drinking water, usually from ponds. A campaign to eradicate the disease was launched in the 1980s and has made significant progress. The strategy of the campaign is discussed, including water supply, health education, case management, and vector control. Current issues including the integration of the campaign into primary health care and the mapping of cases by using geographic information systems are also considered. Finally, some lessons for other disease control and eradication programs are outlined. PMID- 11932233 TI - Integrated approach to malaria control. AB - Malaria draws global attention in a cyclic manner, with interest and associated financing waxing and waning according to political and humanitarian concerns. Currently we are on an upswing, which should be carefully developed. Malaria parasites have been eliminated from Europe and North America through the use of residual insecticides and manipulation of environmental and ecological characteristics; however, in many tropical and some temperate areas the incidence of disease is increasing dramatically. Much of this increase results from a breakdown of effective control methods developed and implemented in the 1960s, but it has also occurred because of a lack of trained scientists and control specialists who live and work in the areas of endemic infection. Add to this the widespread resistance to the most effective antimalarial drug, chloroquine, developing resistance to other first-line drugs such as sulfadoxine pyrimethamine, and resistance of certain vector species of mosquito to some of the previously effective insecticides and we have a crisis situation. Vaccine research has proceeded for over 30 years, but as yet there is no effective product, although research continues in many promising areas. A global strategy for malaria control has been accepted, but there are critics who suggest that the single strategy cannot confront the wide range of conditions in which malaria exists and that reliance on chemotherapy without proper control of drug usage and diagnosis will select for drug resistant parasites, thus exacerbating the problem. An integrated approach to control using vector control strategies based on the biology of the mosquito, the epidemiology of the parasite, and human behavior patterns is needed to prevent continued upsurge in malaria in the endemic areas. PMID- 11932230 TI - Lung infections associated with cystic fibrosis. AB - While originally characterized as a collection of related syndromes, cystic fibrosis (CF) is now recognized as a single disease whose diverse symptoms stem from the wide tissue distribution of the gene product that is defective in CF, the ion channel and regulator, cystic fibrosis transmembrane conductance regulator (CFTR). Defective CFTR protein impacts the function of the pancreas and alters the consistency of mucosal secretions. The latter of these effects probably plays an important role in the defective resistance of CF patients to many pathogens. As the modalities of CF research have changed over the decades from empirical histological studies to include biophysical measurements of CFTR function, the clinical management of this disease has similarly evolved to effectively address the ever-changing spectrum of CF-related infectious diseases. These factors have led to the successful management of many CF-related infections with the notable exception of chronic lung infection with the gram-negative bacterium Pseudomonas aeruginosa. The virulence of P. aeruginosa stems from multiple bacterial attributes, including antibiotic resistance, the ability to utilize quorum-sensing signals to form biofilms, the destructive potential of a multitude of its microbial toxins, and the ability to acquire a mucoid phenotype, which renders this microbe resistant to both the innate and acquired immunologic defenses of the host. PMID- 11932236 TI - New twists on the epigenetics of CpG islands. PMID- 11932237 TI - Functional diversity of Mx proteins: variations on a theme of host resistance to infection. PMID- 11932234 TI - Innate immunity to Mycobacterium tuberculosis. AB - The different manifestations of infection with Mycobacterium tuberculosis reflect the balance between the bacillus and host defense mechanisms. Traditionally, protective immunity to tuberculosis has been ascribed to T-cell-mediated immunity, with CD4(+) T cells playing a crucial role. Recent immunological and genetic studies support the long-standing notion that innate immunity is also relevant in tuberculosis. In this review, emphasis is on these natural, innate host defense mechanisms, referring to experimental data (e.g., studies in gene knockout mice) and epidemiological, immunological, and genetic studies in human tuberculosis. The first step in the innate host defense is cellular uptake of M. tuberculosis, which involves different cellular receptors and humoral factors. Toll-like receptors seem to play a crucial role in immune recognition of M. tuberculosis, which is the next step. The subsequent inflammatory response is regulated by production of pro- and anti-inflammatory cytokines and chemokines. Different natural effector mechanisms for killing of M. tuberculosis have now been identified. Finally, the innate host response is necessary for induction of adaptive immunity to M. tuberculosis. These basic mechanisms augment our understanding of disease pathogenesis and clinical course and will be of help in designing adjunctive treatment strategies. PMID- 11932235 TI - Angiogenic effects of extracellular human immunodeficiency virus type 1 Tat protein and its role in the pathogenesis of AIDS-associated Kaposi's sarcoma. AB - The Tat protein of human immunodeficiency virus (HIV) type 1 is a transactivator of viral gene expression that is required for virus replication and spread. Moreover, Tat is released by acutely HIV-infected cells via a leaderless secretory pathway and in a biologically active form that exerts effects on both HIV-infected and uninfected cells from different organs and systems. This review focuses on the activities of extracellular Tat protein on endothelial cells, on angiogenesis, and on the pathogenesis of AIDS-associated angioproliferative diseases such as Kaposi's sarcoma. In particular, we discuss results from different groups indicating that Tat mimics the proangiogenic activities of extracellular matrix molecules and that it enhances the effects of angiogenic factors. PMID- 11932232 TI - Genotypic testing for human immunodeficiency virus type 1 drug resistance. AB - There are 16 approved human immunodeficiency virus type 1 (HIV-1) drugs belonging to three mechanistic classes: protease inhibitors, nucleoside and nucleotide reverse transcriptase (RT) inhibitors, and nonnucleoside RT inhibitors. HIV-1 resistance to these drugs is caused by mutations in the protease and RT enzymes, the molecular targets of these drugs. Drug resistance mutations arise most often in treated individuals, resulting from selective drug pressure in the presence of incompletely suppressed virus replication. HIV-1 isolates with drug resistance mutations, however, may also be transmitted to newly infected individuals. Three expert panels have recommended that HIV-1 protease and RT susceptibility testing should be used to help select HIV drug therapy. Although genotypic testing is more complex than typical antimicrobial susceptibility tests, there is a rich literature supporting the prognostic value of HIV-1 protease and RT mutations. This review describes the genetic mechanisms of HIV-1 drug resistance and summarizes published data linking individual RT and protease mutations to in vitro and in vivo resistance to the currently available HIV drugs. PMID- 11932238 TI - The genome of M. acetivorans reveals extensive metabolic and physiological diversity. AB - Methanogenesis, the biological production of methane, plays a pivotal role in the global carbon cycle and contributes significantly to global warming. The majority of methane in nature is derived from acetate. Here we report the complete genome sequence of an acetate-utilizing methanogen, Methanosarcina acetivorans C2A. Methanosarcineae are the most metabolically diverse methanogens, thrive in a broad range of environments, and are unique among the Archaea in forming complex multicellular structures. This diversity is reflected in the genome of M. acetivorans. At 5,751,492 base pairs it is by far the largest known archaeal genome. The 4524 open reading frames code for a strikingly wide and unanticipated variety of metabolic and cellular capabilities. The presence of novel methyltransferases indicates the likelihood of undiscovered natural energy sources for methanogenesis, whereas the presence of single-subunit carbon monoxide dehydrogenases raises the possibility of nonmethanogenic growth. Although motility has not been observed in any Methanosarcineae, a flagellin gene cluster and two complete chemotaxis gene clusters were identified. The availability of genetic methods, coupled with its physiological and metabolic diversity, makes M. acetivorans a powerful model organism for the study of archaeal biology. [Sequence, data, annotations and analyses are available at http://www-genome.wi.mit.edu/.] PMID- 11932239 TI - A genome-wide screen for normally methylated human CpG islands that can identify novel imprinted genes. AB - DNA methylation is a covalent modification of the nucleotide cytosine that is stably inherited at the dinucleotide CpG by somatic cells, and 70% of CpG dinucleotides in the genome are methylated. The exception to this pattern of methylation are CpG islands, CpG-rich sequences that are protected from methylation, and generally are thought to be methylated only on the inactive X chromosome and in tumors, as well as differentially methylated regions (DMRs) in the vicinity of imprinted genes. To identify chromosomal regions that might harbor imprinted genes, we devised a strategy for isolating a library of normally methylated CpG islands. Most of the methylated CpG islands represented high copy number dispersed repeats. However, 62 unique clones in the library were characterized, all of which were methylated and GC-rich, with a GC content >50%. Of these, 43 clones also showed a CpG(obs)/CpG(exp) >0.6, of which 30 were studied in detail. These unique methylated CpG islands mapped to 23 chromosomal regions, and 12 were differentially methylated regions in uniparental tissues of germline origin, i.e., hydatidiform moles (paternal origin) and complete ovarian teratomas (maternal origin), even though many apparently were methylated in somatic tissues. We term these sequences gDMRs, for germline differentially methylated regions. At least two gDMRs mapped near imprinted genes, HYMA1 and a novel homolog of Elongin A and Elongin A2, which we term Elongin A3. Surprisingly, 18 of the methylated CpG islands were methylated in germline tissues of both parental origins, representing a previously uncharacterized class of normally methylated CpG islands in the genome, and which we term similarly methylated regions (SMRs). These SMRs, in contrast to the gDMRs, were significantly associated with telomeric band locations (P =.0008), suggesting a potential role for SMRs in chromosome organization. At least 10 of the methylated CpG islands were on average 85% conserved between mouse and human. These sequences will provide a valuable resource in the search for novel imprinted genes, for defining the molecular substrates of the normal methylome, and for identifying novel targets for mammalian chromatin formation. PMID- 11932241 TI - Comparative analysis of chloroplast genomes: functional annotation, genome-based phylogeny, and deduced evolutionary patterns. AB - All protein sequences from 19 complete chloroplast genomes (cpDNA) have been studied using a new computational method able to analyze functional correlations among series of protein sequences contained in complete proteomes. First, all open reading frames (ORFs) from the cpDNAs, comprising a total of 2266 protein sequences, were compared against the 3168 proteins from Synechocystis PCC6803 complete genome to find functionally related orthologous proteins. Additionally, all cpDNA genomes were pairwise compared to find orthologous groups not present in cyanobacteria. Annotations in the cluster of othologous proteins database and CyanoBase were used as reference for the functional assignments. Following this protocol, new functional assignments were made for ORFs of unknown function and for ycfs (hypothetical chloroplast frames), which still lack a functional assignment. Using this information, a matrix of functional relationships was derived from profiles of the presence and/or absence of orthologous proteins; the matrix included 1837 proteins in 277 orthologous clusters. A factor analysis study of this matrix, followed by cluster analysis, allowed us to obtain accurate phylogenetic reconstructions and the detection of genes probably involved in speciation as phylogenetic correlates. Finally, by grouping common evolutionary patterns, we show that it is possible to determine functionally linked protein networks. This has allowed us to suggest putative associations for some unknown ORFs. PMID- 11932242 TI - Functional genomics on potato virus A: virus genome-wide map of sites essential for virus propagation. AB - Transposition-based in vitro insertional mutagenesis strategies provide promising new approaches for functional characterization of any cloned gene or genome region. We have extended the methodology and scope of such analysis to a complete viral genome. To map genome regions both essential and nonessential for Potato virus A propagation, we generated a genomic 15-bp insertion mutant library utilizing the efficient in vitro DNA transposition reaction of phage Mu. We then determined the proficiency of 1125 mutants to propagate in tobacco protoplasts by using a genetic footprinting strategy that simultaneously mapped the genomic insertion sites. Over 300 sites critical for virus propagation were identified, and many of them were located in positions previously not assigned to any viral functions. Many genome regions tolerated insertions indicating less important sites for virus propagation and thus pinpointed potential locations for further genome manipulation. The methodology described is applicable to a detailed functional analysis of any viral nucleic acid cloned as DNA and can be used to address many different processes during viral infection cycles. PMID- 11932243 TI - Polymorphisms and the differential antiviral activity of the chicken Mx gene. AB - The nucleotide sequence of chicken Mx cDNA was reported earlier using the White Leghorn breed in Germany, but it showed no enhanced resistance to viruses. In this study, the nucleotide sequences of chicken Mx cDNA were determined in many breeds. A total of 25 nucleotide substitutions, of which 14 were deduced to cause amino acid exchanges, were detected, suggesting that the chicken Mx gene is very polymorphic. Transfected cell clones expressing chicken Mx mRNA were established after the Mx cDNA was constructed with an expression vector and introduced into mouse 3T3 cells, and the Mx genes from some breeds were demonstrated to confer positive antiviral responses to influenza virus and vesicular stomatitis virus. On the basis of the comparison among the antiviral activities associated with many Mx variations, a specific amino acid substitution at position 631 (Ser to Asn) was considered to determine the antivirally positive or negative Mx gene. Thus, a single amino acid substitution influences the antiviral activity of Mx in domesticated chickens. PMID- 11932244 TI - Patterns of human diversity, within and among continents, inferred from biallelic DNA polymorphisms. AB - Previous studies have reported that about 85% of human diversity at Short Tandem Repeat (STR) and Restriction Fragment Length Polymorphism (RFLP) autosomal loci is due to differences between individuals of the same population, whereas differences among continental groups account for only 10% of the overall genetic variance. These findings conflict with popular notions of distinct and relatively homogeneous human races, and may also call into question the apparent usefulness of ethnic classification in, for example, medical diagnostics. Here, we present new data on 21 Alu insertions in 32 populations. We analyze these data along with three other large, globally dispersed data sets consisting of apparently neutral biallelic nuclear markers, as well as with a beta-globin data set possibly subject to selection. We confirm the previous results for the autosomal data, and find a higher diversity among continents for Y-chromosome loci. We also extend the analyses to address two questions: (1) whether differences between continental groups, although small, are nevertheless large enough to confidently assign individuals to their continent on the basis of their genotypes; (2) whether the observed genotypes naturally cluster into continental or population groups when the sample source location is ignored. Using a range of statistical methods, we show that classification errors are at best around 30% for autosomal biallelic polymorphisms and 27% for the Y chromosome. Two data sets suggest the existence of three and four major groups of genotypes worldwide, respectively, and the two groupings are inconsistent. These results suggest that, at random biallelic loci, there is little evidence, if any, of a clear subdivision of humans into biologically defined groups. PMID- 11932245 TI - Vestibular dysfunction in the epistatic circler mouse is caused by phenotypic interaction of one recessive gene and three modifier genes. AB - Vestibular dysfunction is a frequent clinical problem, leading to dizziness and imbalance. Genes play an important role in its etiology, but the genetics are complex and poorly understood. In this study we have analyzed the complex inheritance pattern in the Epistatic circler mouse, which shows circling behavior indicative of vestibular dysfunction in the mouse. This phenotype exists in a proportion of the F2-generation from an intercross between C57L/J and SWR/J mouse strains. Genetic investigation indicates that the circling behavior is caused by a major recessively inherited gene derived from the SWR/J strain (the Ecs-gene) in combination with at least three different modifier genes derived from C57L/J (the Ecl-genes). Genetic mapping made it possible to localize the Ecs-gene to chromosome 14 and the Ecl-genes to chromosome 3, 4, and 13. This study illustrates the feasibility of identifying genes for multifactorial traits in mice. PMID- 11932246 TI - A quantitative trait locus influencing anxiety in the laboratory rat. AB - A critical test for a gene that influences susceptibility to fear in animals is that it should have a consistent pattern of effects across a broad range of conditioned and unconditioned models of anxiety. Despite many years of research, definitive evidence that genetic effects operate in this way is lacking. The limited behavioral test regimes so far used in genetic mapping experiments and the lack of suitable multivariate methodologies have made it impossible to determine whether the quantitative trait loci (QTL) detected to date specifically influence fear-related traits. Here we report the first multivariate analysis to explore the genetic architecture of rodent behavior in a battery of animal models of anxiety. We have mapped QTLs in an F2 intercross of two rat strains, the Roman high and low avoidance rats, that have been selectively bred for differential response to fear. Multivariate analyses show that one locus, on rat chromosome 5, influences behavior in different models of anxiety. The QTL influences two-way active avoidance, conditioned fear, elevated plus maze, and open field activity but not acoustic startle response or defecation in a novel environment. The direction of effects of the QTL alleles and a coincidence between the behavioral profiles of anxiolytic drug and genetic action are consistent with the QTL containing at least one gene with a pleiotropic action on fear responses. As the neural basis of fear is conserved across species, we suggest that the QTL may have relevance to trait anxiety in humans. PMID- 11932240 TI - Annotated expressed sequence tags and cDNA microarrays for studies of brain and behavior in the honey bee. AB - To accelerate the molecular analysis of behavior in the honey bee (Apis mellifera), we created expressed sequence tag (EST) and cDNA microarray resources for the bee brain. Over 20,000 cDNA clones were partially sequenced from a normalized (and subsequently subtracted) library generated from adult A. mellifera brains. These sequences were processed to identify 15,311 high-quality ESTs representing 8912 putative transcripts. Putative transcripts were functionally annotated (using the Gene Ontology classification system) based on matching gene sequences in Drosophila melanogaster. The brain ESTs represent a broad range of molecular functions and biological processes, with neurobiological classifications particularly well represented. Roughly half of Drosophila genes currently implicated in synaptic transmission and/or behavior are represented in the Apis EST set. Of Apis sequences with open reading frames of at least 450 bp, 24% are highly diverged with no matches to known protein sequences. Additionally, over 100 Apis transcript sequences conserved with other organisms appear to have been lost from the Drosophila genome. DNA microarrays were fabricated with over 7000 EST cDNA clones putatively representing different transcripts. Using probe derived from single bee brain mRNA, microarrays detected gene expression for 90% of Apis cDNAs two standard deviations greater than exogenous control cDNAs. [The sequence data described in this paper have been submitted to Genbank data library under accession nos. BI502708-BI517278. The sequences are also available at http://titan.biotec.uiuc.edu/bee/honeybee_project.htm.] PMID- 11932247 TI - Complex SNP-based haplotypes in three human helicases: implications for cancer association studies. AB - We have initiated a candidate gene approach to study variation and predisposition to cancer in the four major ethnic groups that constitute the U.S. population (African Americans, Caucasians, Hispanics, and Asians). We resequenced portions of three helicase genes (BLM, WRN, and RECQL) identifying a total of 37 noncoding single nucleotide polymorphisms (SNPs). Haplotype inference predicted 50 haplotypes in BLM, 56 in WRN, and 47 in RECQL in a sample of 600 chromosomes. Approximately 10% of the predicted haplotypes were shared among all ethnic groups. Linkage disequilibrium and recombination effects showed that each locus has taken a diverse evolutionary path. Primate DNA analysis of the same loci revealed one human haplotype per gene shared with the great apes, indicating that the observed diversity occurred since the divergence of humans from the last common ancestor. In BLM, we confirmed the presence of a founder haplotype among Ashkenazi Jews homozygous for the blm(Ash) mutation. The cosegregating haplotype was seen in all (6/6) samples of Ashkenazi descent, whereas in the general population it has a low frequency (0.02) and was not found in African Americans. In WRN, ethnic samples were studied for their haplotype content and the presence or absence of six previously described coding SNPs (cSNPs). Hispanic individuals carrying two of these cSNPs showed a 60% increase in the frequency of a common haplotype (haplotype No. 28). In the pooled sample, no association was found. Because (1) the majority of the haplotypes are population specific and (2) the patterns of linkage disequilibrium, recombination, and haplotype diversity are markedly different between gene regions, these data show the importance of either ethnically matched controls or within-family-based disease-gene association studies. PMID- 11932248 TI - Engineering a reduced Escherichia coli genome. AB - Our goal is to construct an improved Escherichia coli to serve both as a better model organism and as a more useful technological tool for genome science. We developed techniques for precise genomic surgery and applied them to deleting the largest K-islands of E. coli, identified by comparative genomics as recent horizontal acquisitions to the genome. They are loaded with cryptic prophages, transposons, damaged genes, and genes of unknown function. Our method leaves no scars or markers behind and can be applied sequentially. Twelve K-islands were successfully deleted, resulting in an 8.1% reduced genome size, a 9.3% reduction of gene count, and elimination of 24 of the 44 transposable elements of E. coli. These are particularly detrimental because they can mutagenize the genome or transpose into clones being propagated for sequencing, as happened in 18 places of the draft human genome sequence. We found no change in the growth rate on minimal medium, confirming the nonessential nature of these islands. This demonstration of feasibility opens the way for constructing a maximally reduced strain, which will provide a clean background for functional genomics studies, a more efficient background for use in biotechnology applications, and a unique tool for studies of genome stability and evolution. PMID- 11932249 TI - Predicting gene ontology functions from ProDom and CDD protein domains. AB - A heuristic algorithm for associating Gene Ontology (GO) defined molecular functions to protein domains as listed in the ProDom and CDD databases is described. The algorithm generates rules for function-domain associations based on the intersection of functions assigned to gene products by the GO consortium that contain ProDom and/or CDD domains at varying levels of sequence similarity. The hierarchical nature of GO molecular functions is incorporated into rule generation. Manual review of a subset of the rules generated indicates an accuracy rate of 87% for ProDom rules and 84% for CDD rules. The utility of these associations is that novel sequences can be assigned a putative function if sufficient similarity exists to a ProDom or CDD domain for which one or more GO functions has been associated. Although functional assignments are increasingly being made for gene products from model organisms, it is likely that the needs of investigators will continue to outpace the efforts of curators, particularly for nonmodel organisms. A comparison with other methods in terms of coverage and agreement was performed, indicating the utility of the approach. The domain function associations and function assignments are available from our website http://www.cbil.upenn.edu/GO. PMID- 11932250 TI - BLAT--the BLAST-like alignment tool. AB - Analyzing vertebrate genomes requires rapid mRNA/DNA and cross-species protein alignments. A new tool, BLAT, is more accurate and 500 times faster than popular existing tools for mRNA/DNA alignments and 50 times faster for protein alignments at sensitivity settings typically used when comparing vertebrate sequences. BLAT's speed stems from an index of all nonoverlapping K-mers in the genome. This index fits inside the RAM of inexpensive computers, and need only be computed once for each genome assembly. BLAT has several major stages. It uses the index to find regions in the genome likely to be homologous to the query sequence. It performs an alignment between homologous regions. It stitches together these aligned regions (often exons) into larger alignments (typically genes). Finally, BLAT revisits small internal exons possibly missed at the first stage and adjusts large gap boundaries that have canonical splice sites where feasible. This paper describes how BLAT was optimized. Effects on speed and sensitivity are explored for various K-mer sizes, mismatch schemes, and number of required index matches. BLAT is compared with other alignment programs on various test sets and then used in several genome-wide applications. http://genome.ucsc.edu hosts a web-based BLAT server for the human genome. PMID- 11932251 TI - Ran-binding protein 3 links Crm1 to the Ran guanine nucleotide exchange factor. AB - Ran-binding protein 3 (RanBP3) is an approximately 55-kDa protein that functions as a cofactor for Crm1-mediated nuclear export. RanBP3 stimulates export by enhancing the affinity of Crm1 for Ran.GTP and cargo. However, important additional functions for this cofactor may exist. We now report that RanBP3 associates with the Ran-specific guanine nucleotide exchange factor, regulator of chromosome condensation 1 (RCC1). This interaction was stimulated by the addition of Ran; moreover, Ran.GDP, Ran.GTP, and Ran without nucleotide could all stimulate complex formation between RanBP3 and RCC1 even though binding of Ran.GDP to RanBP3 alone was undetectable. RanBP3 could also promote binding of Crm1 to RCC1 in the presence of Ran. Binding of RanBP3 to RCC1 increased the catalytic activity of RCC1 toward Ran, and importantly, the ability of RanBP3 to stimulate RCC1 was not affected by the presence of Crm1. These data indicate that RanBP3 acts as a scaffold protein to promote the efficient assembly of export complexes. By tethering Crm1 to catalytically enhanced RCC1, RanBP3 may lower the entropic barrier for the loading of Ran.GTP onto Crm1. We propose that this provides an additional mechanism by which RanBP3 facilitates export. PMID- 11932252 TI - Cleavage of the carboxyl tail from the G3 domain of aggrecan but not versican and identification of the amino acids involved in the degradation. AB - Aggrecan, a major structural proteoglycan in cartilage, contains three globular domains, G1, G2, and G3, as well as sequences for glycosaminoglycan modification. A large number of proteases are implicated in aggrecan cleavage in normal metabolism, aging, and arthritis. These proteases are known to cleave at the IGD, KS, and CS domains. Here we report for the first time evidence of cleavage at a novel site, the carboxyl tail of aggrecan. Results from deletion mutants of the tail indicated that the likely cleavage sites were two consensus sequences, RRLXK and RSPR, present in the aggrecan analogs of many species. This was confirmed by site-directed mutagenesis. A construct containing two G3 domains (G3G3) was also found to cleave between the G3 duplicates. When G3 tail was linked to a glycosaminoglycan-modifying sequence, it was protected from cleavage. Furin inhibitor also reduced the levels of tail cleavage. The carboxyl tails of chicken and human versican were not cleaved, despite the presence of the consensus sequence. Our studies indicate that the basic amino acids present in the tail play an important role in cleavage, and this mechanism is specific to aggrecan. PMID- 11932253 TI - Functional consequences of the loss of high affinity agonist binding to gamma aminobutyric acid type A receptors. Implications for receptor desensitization. AB - We reported previously that tyrosine 62 of the beta2 subunit of the gamma aminobutyric acid, type A (GABA(A)) receptor is an important determinant of high affinity agonist binding and that recombinant alpha1beta2gamma2(L) receptors carrying the Y62S mutation lack measurable high affinity sites for [3H]muscimol. We have now examined the effects of disrupting these sites on the macroscopic desensitization properties of receptors expressed in Xenopus oocytes. Desensitization was measured by the ability of low concentrations of bath perfused agonist to reduce the current responses elicited by subsequent challenges with saturating concentrations of GABA. Wild-type receptors were desensitized by pre-perfused muscimol with an IC50 approximately 0.7 microm, which correlates well with the lower affinity sites for this agonist that are measured in direct binding studies. Receptors carrying the beta2 Y62S and Y62F mutations desensitized at slightly higher (2-7-fold) agonist concentrations. However, at low perfusate concentrations, the Y62S-containing receptor recovered from the desensitized state even in the continued presence of agonist. The characteristics of desensitization in the wild-type and mutant receptors lead us to suggest that the major role of the high affinity agonist-binding site(s) of the GABA(A) receptor is not to induce desensitization but rather to stabilize the desensitized state once it has been formed. PMID- 11932254 TI - Inositol polyphosphate 1-phosphatase is a novel antihypertrophic factor. AB - Activation of G(q)-coupled alpha(1)-adrenergic receptors leads to hypertrophic growth of neonatal rat ventricular cardiomyocytes that is associated with increased expression of hypertrophy-related genes, including atrial natriuretic peptide (ANP) and myosin light chain-2 (MLC), as well as increased ribosome synthesis. The role of inositol phosphates in signaling pathways involved in these changes in gene expression was examined by overexpressing inositol phosphate-metabolizing enzymes and determining effects on ANP, MLC, and 45 S ribosomal gene expression following co-transfection of appropriate reporter gene constructs. Overexpression of enzymes that metabolize inositol 1,4,5 trisphosphate did not reduce ANP or MLC responses, but overexpression of the enzyme primarily responsible for metabolism of inositol 4,5-bisphosphate (Ins(1,4)P(2)), inositol polyphosphate 1-phosphatase (INPP), reduced ANP and MLC responses associated with alpha(1)-adrenergic receptor-mediated hypertrophy. Similarly overexpressed INPP reduced ANP and MLC responses associated with contraction-induced hypertrophy. In addition, overexpression of INPP reduced the increase in ribosomal DNA transcription associated with both hypertrophic models. Hypertrophied cells from both cell models as well as ventricular tissue from mouse hearts hypertrophied by pressure overload in vivo contained heightened levels of Ins(1,4)P(2), suggesting reduced INPP activity in three different models of hypertrophy. These studies provide evidence for an involvement of Ins(1,4)P(2) in hypertrophic signaling pathways in ventricular myocytes. PMID- 11932255 TI - The phosphotyrosine binding-like domain of talin activates integrins. AB - Cellular regulation of the ligand binding affinity of integrin adhesion receptors (integrin activation) depends on the integrin beta cytoplasmic domains (tails). The head domain of talin binds to several integrin beta tails and activates integrins. This head domain contains a predicted FERM domain composed of three subdomains (F1, F2, and F3). An integrin-activating talin fragment was predicted to contain the F2 and F3 subdomains. Both isolated subdomains bound specifically to the integrin beta3 tail. However, talin F3 bound the beta3 tail with a 4-fold higher affinity than talin F2. Furthermore, expression of talin F3 (but not F2) in cells led to activation of integrin alpha(IIb)beta3. A molecular model of talin F3 indicated that it resembles a phosphotyrosine-binding (PTB) domain. PTB domains recognize peptide ligands containing beta turns, often formed by NPXY motifs. NPX(Y/F) motifs are highly conserved in integrin beta tails, and mutations that disrupt this motif interfere with both integrin activation and talin binding. Thus, integrin binding to talin resembles the interactions of PTB domains with peptide ligands. These resemblances suggest that the activation of integrins requires the presence of a beta turn at NPX(Y/F) motifs conserved in integrin beta cytoplasmic domains. PMID- 11932256 TI - Savignygrin, a platelet aggregation inhibitor from the soft tick Ornithodoros savignyi, presents the RGD integrin recognition motif on the Kunitz-BPTI fold. AB - Savignygrin, a platelet aggregation inhibitor that possesses the RGD integrin recognition motif, has been purified from the soft tick Ornithodoros savignyi. Two isoforms with similar biological activities differ because of R52G and N60G in their amino acid sequences, indicating a recent gene duplication event. Platelet aggregation induced by ADP (IC50, 130 nm), collagen, the thrombin receptor-activating peptide, and epinephrine was inhibited, although platelets were activated and underwent a shape change. The binding of alpha-CD41 (P2) to platelets, the binding of purified alpha(IIb)beta3 to fibrinogen, and the adhesion of platelets to fibrinogen was inhibited, indicating a targeting of the fibrinogen receptor. In contrast, the adhesion of osteosarcoma cells that express the integrin alpha(v)beta3 to vitronectin or fibrinogen was not inhibited, indicating the specificity of savignygrin toward alpha(IIb)beta3. Savignygrin shows sequence identity to disagregin, a platelet aggregation inhibitor from the tick Ornithodoros moubata that lacks an RGD motif. The cysteine arrangement of savignygrin is similar to that of the bovine pancreatic trypsin inhibitor family of serine protease inhibitors. A homology model based on the structure of the tick anticoagulant peptide indicates that the RGD motif is presented on the substrate-binding loop of the canonical BPTI inhibitors. However, savignygrin did not inhibit the serine proteases fXa, plasmin, thrombin, or trypsin. This is the first report of a platelet aggregation inhibitor that presents the RGD motif using the Kunitz-BPTI protein fold. PMID- 11932257 TI - Presence of cathepsin B in the human pancreatic secretory pathway and its role in trypsinogen activation during hereditary pancreatitis. AB - The lysosomal cysteine protease cathepsin B is thought to play a central role in intrapancreatic trypsinogen activation and the onset of experimental pancreatitis. Recent in vitro studies have suggested that this mechanism might be of pathophysiological relevance in hereditary pancreatitis, a human inborn disorder associated with mutations in the cationic trypsinogen gene. In the present study evidence is presented that cathepsin B is abundantly present in the secretory compartment of the human exocrine pancreas, as judged by immunogold electron microscopy. Moreover, pro-cathepsin B and mature cathepsin B are both secreted together with trypsinogen and active trypsin into the pancreatic juice of patients with sporadic pancreatitis or hereditary pancreatitis. Finally, cathepsin B- catalyzed activation of recombinant human cationic trypsinogen with hereditary pancreatitis-associated mutations N29I, N29T, or R122H were characterized. In contrast to a previous report, cathepsin B-mediated activation of wild type and all three mutant trypsinogen forms was essentially identical under a wide range of experimental conditions. These observations confirm the presence of active cathepsin B in the human pancreatic secretory pathway and are consistent with the notion that cathepsin B-mediated trypsinogen activation might play a pathogenic role in human pancreatitis. On the other hand, the results clearly demonstrate that hereditary pancreatitis-associated mutations do not lead to increased or decreased trypsinogen activation by cathepsin B. Therefore, mutation-dependent alterations in cathepsin B-induced trypsinogen activation are not the cause of hereditary pancreatitis. PMID- 11932258 TI - Polylysine induces an antiparallel actin dimer that nucleates filament assembly: crystal structure at 3.5-A resolution. AB - An antiparallel actin dimer has been proposed to be an intermediate species during actin filament nucleation. We now show that latrunculin A, a marine natural product that inhibits actin polymerization, arrests polylysine-induced nucleation at the level of an antiparallel dimer, resulting in its accumulation. These dimers, when composed of pyrene-labeled actin subunits, give rise to a fluorescent excimer, permitting detection during polymerization in vitro. We report the crystallographic structure of the polylysine-actin-latrunculin A complex at 3.5-A resolution. The non-crystallographic contact is consistent with a dimeric structure and confirms the antiparallel orientation of its subunits. The crystallographic contacts reveal that the mobile DNase I binding loop of one subunit of a symmetry-related antiparallel actin dimer is partially stabilized in the interface between the two subunits of a second antiparallel dimer. These results provide a potential explanation for the paradoxical nucleation of actin filaments that have exclusively parallel subunits by a dimer containing antiparallel subunits. PMID- 11932259 TI - Insertion of bitopic membrane proteins into the inner membrane of mitochondria involves an export step from the matrix. AB - The mitochondrial inner membrane contains a large number of polytopic proteins that are derived from prokaryotic ancestors of mitochondria. Little is known about the intramitochondrial sorting of these proteins. We chose two proteins of known topology as examples to study the pathway of insertion into the inner membrane; Mrs2 and Yta10 are bitopic proteins that expose negatively charged loops of different complexity into the intermembrane space. Here we show that both Mrs2 and Yta10 transiently accumulate as sorting intermediates in the matrix before they integrate into the inner membrane. The sorting pathway of both proteins can be separated into two sequential reactions: (i) import into the matrix and (ii) insertion from the matrix into the inner membrane. The latter process was found to depend on the membrane potential and, in this respect, is similar to the insertion of membrane proteins in bacteria. A comparison of the charge distribution of intermembrane space loops in a variety of mitochondrial inner membrane proteins suggests that this mode of "conservative sorting" might be the typical insertion route for polytopic inner membrane proteins that originated from bacterial ancestors. PMID- 11932260 TI - Expression and localization of aquaporin water channels in rat hepatocytes. Evidence for a role in canalicular bile secretion. AB - Although bile formation requires that large volumes of water be rapidly transported across liver epithelia, including hepatocytes, the molecular mechanisms by which water is secreted into bile are obscure. The aquaporins are a family of 10 channel-forming, integral membrane proteins of approximately 28 kDa numbered 0-9 that allow water to rapidly traverse epithelial barriers in several organs including kidney, eye, and brain. We found transcripts of three of 10 aquaporins in hepatocytes (aquaporin 8 aquaporin 9 > aquaporin 0) by reverse transcription-polymerase chain reaction and quantitative ribonuclease protection assays; immunohistochemistry confirmed the presence of these three proteins in liver. Immunoblots of subcellular fractions of hepatocytes showed enrichment of aquaporins 0 and 8 in microsomes and canalicular plasma membranes; aquaporin 9 was enriched only in basolateral plasma membranes. Immunofluorescence of hepatocyte couplets confirmed the intracellular/canalicular localization of aquaporins 0 and 8 and the basolateral localization of aquaporin 9. Upon exposure of couplets to a choleretic stimulus (i.e. dibutyryl cAMP), aquaporin 8 redistributed to the canalicular plasma membrane; the subcellular distributions of aquaporins 0 and 9 were unaffected. In addition, exposure of couplets to dibutyryl cAMP caused an increase in canalicular water transport in the presence and absence of an osmotic gradient, an effect that was blocked by aquaporin inhibitors. These results provide evidence that aquaporins are present in hepatocytes and that aquaporins are involved in agonist-stimulated canalicular bile secretion. PMID- 11932261 TI - Targeted gene disruption reveals the role of cysteinyl leukotriene 1 receptor in the enhanced vascular permeability of mice undergoing acute inflammatory responses. AB - The cysteinyl leukotrienes (cysLTs), leukotriene (LT) C(4), LTD(4), and LTE(4), are proinflammatory lipid mediators generated in the mouse by hematopoietic cells such as macrophages and mast cells. There are two mouse receptors for the cysLTs, CysLT(1) receptor (CysLT(1)R) and CysLT(2)R, which are 38% homologous and are located on mouse chromosomes X and 14, respectively. To clarify the different roles of the CysLT(1)R and CysLT(2)R in inflammatory responses in vivo, we generated CysLT(1)R-deficient mice by targeted gene disruption. These mice developed normally and were fertile. In an intracellular calcium mobilization assay with fura-2 acetoxymethyl ester, peritoneal macrophages from wild-type littermates, which express both CysLT(1)R and CysLT(2)R, responded substantially to 1 x 10(-6) m LTD(4) and slightly to 1 x 10(-6) m LTC(4), whereas the macrophages from CysLT(1)R-deficient mice did not respond to either LTD(4) or LTC(4). Plasma protein extravasation, but not neutrophil infiltration, was significantly reduced in CysLT(1)R-deficient mice subjected to zymosan A-induced peritoneal inflammation. Plasma protein extravasation was also significantly diminished in CysLT(1)R-deficient mice undergoing IgE-mediated passive cutaneous anaphylaxis as compared with the wild-type mice. Thus, the cysLTs generated in vivo by either monocytes/macrophages or mast cells utilize CysLT(1)R for the response of the microvasculature in acute inflammation. PMID- 11932262 TI - Clinical review 144: Estrogen and the male skeleton. AB - Because estrogen (E) and T are the major sex steroids in women and men, respectively, the traditional view had been that E primarily regulated bone turnover in women and T played the analogous role in men. The description of ER- deficient and aromatase-deficient males, however, initiated a major shift in our thinking on the relative roles of T and E in regulating the male skeleton, because these individuals all had unfused epiphyses, high bone turnover, and osteopenia. Similar, albeit less striking, findings were noted in mouse models with knock-out of either the ER-alpha or the aromatase genes. Although these human experiments of nature and mouse knock-out models clearly demonstrated an important role for E in the growth and maturation of the male skeleton, they did not define the role of E vs. T in regulating the adult male skeleton. The past several years have witnessed an accumulation of evidence from observational as well as direct interventional studies that now clearly indicates that E plays a major, and likely dominant, role in bone metabolism in men. These data also suggest that a threshold level of bioavailable (or non-SHBG bound) E is needed for skeletal E sufficiency in the male, and that with aging, an increasing percentage of elderly men begin to fall below this level. It is this subset of men who may be at greatest risk for the development of age-related bone loss and osteoporosis. Moreover, these men may also be the ones most likely to respond favorably to treatment with selective E receptor modulators, or perhaps even to T replacement, because the skeletal effects of the latter may be mediated largely via aromatization to E. PMID- 11932263 TI - Clinical review 145: Pleiotropic effects of statins: lipid reduction and beyond. AB - There is accumulating evidence that statins have beneficial effects that are independent of their classical actions on lipoproteins. These effects include reductions in inflammation in the vasculature, kidney, and bone. Potential beneficial effects of these agents include enhancement of nitric oxide production in vasculature and the kidney. These agents appear to reduce bone fractures and may improve insulin sensitivity and reduce the likelihood of persons progressing from impaired glucose tolerance to type II diabetes. Potential beneficial pleiotropic effects of statins are covered in this review. PMID- 11932264 TI - A pathologist trying to help endocrinologists to interpret cytopathology reports from thyroid aspirates. AB - Fine-needle aspiration has become the diagnostic tool of choice in the initial evaluation of thyroid nodules. As a cytopathologist who has been performing fine needle aspirations for over 25 yr at a university hospital and for the last 3 yr at the largest not-for-profit hospital in the Washington, D.C. area, I have seen increasing demand for this service. However, there is also discontent with the cytopathology reports issued by numerous laboratories. Pathologists have to standardize and simplify their reports so that they will be more meaningful to the referring physician. Endocrinologists should communicate their needs to the pathologists and insist on clear, succinct, and prompt diagnoses. PMID- 11932265 TI - Should the nonaromatizable androgen dihydrotestosterone be considered as an alternative to testosterone in the treatment of the andropause? PMID- 11932266 TI - The effects of transdermal dihydrotestosterone in the aging male: a prospective, randomized, double blind study. AB - The objective of the study was to investigate the effects of dihydrotestosterone (DHT) gel on general well-being, sexual function, and the prostate in aging men. A total of 120 men participated in this randomized, placebo-controlled study (60 DHT and 60 placebo). All subjects had nocturnal penile tumescence once per week or less, andropause symptoms, and a serum T level of 15 nmol/liter or less and/or a serum SHBG level greater than 30 nmol/liter. The mean age was 58 yr (range, 50 70 yr). Of these subjects, 114 men completed the study. DHT was administered transdermally for 6 months, and the dose varied from 125-250 mg/d. General well being symptoms and sexual function were evaluated using a questionnaire, and prostate symptoms were evaluated using the International Prostate Symptoms Score, transrectal ultrasonography, and assay of serum prostate-specific antigen. Early morning erections improved transiently in the DHT group at 3 months of treatment (P < 0.003), and the ability to maintain erection improved in the DHT group compared with the placebo group (P < 0.04). No significant changes were observed in general well-being between the placebo and the DHT group. Serum concentrations of LH, FSH, E2, T, and SHBG decreased significantly during DHT treatment. Treatment with DHT did not affect liver function or the lipid profile. Hemoglobin concentrations increased from 146.0 +/- 8.2 to 154.8 +/- 11.4 g/liter, and hematocrit from 43.5 +/- 2.5% to 45.8 +/- 3.4% (P < 0.001). Prostate weight and prostate-specific antigen levels did not change during the treatment. No major adverse events were observed. Transdermal administration of DHT improves sexual function and may be a useful alternative for androgen replacement. As estrogens are thought to play a role in the pathogenesis of prostate hyperplasia, DHT may be beneficial, compared with aromatizing androgens, in the treatment of aging men. PMID- 11932267 TI - The genetics of absorptive hypercalciuria--a note of caution. PMID- 11932268 TI - Identification and characterization of a gene with base substitutions associated with the absorptive hypercalciuria phenotype and low spinal bone density. AB - Absorptive hypercalciuria (AH) is a kidney stone-forming condition frequently complicated by bone loss. Previously, we mapped the locus for an inherited form of AH to chromosome 1q23.3-q24. We have sequenced a putative gene (subsequently shown by others to be homologous with the rat soluble adenylate cyclase gene) in this region in 12 unrelated Caucasian AH patients. Eighteen base substitutions were identified in the soluble adenylate cyclase human homolog gene. All sequence variations were further evaluated in 3-68 additional unrelated AH patients and 19 132 normal subjects, and 1 additional base substitution was identified. Six of the identified sequence variations occurred with increased frequency in the AH population and tracked with the AH phenotype in AH families. Calculated odds ratios showed that the occurrence of any 4 of these individual base substitutions was associated with a 2.2- to 3.5-fold increase in estimated risk for AH (P < 0.02). In addition, 1 or more base changes was associated with a lower L2-L4 vertebral bone density. Sequence analysis of 3 other genes within the AH linkage interval showed no difference in the distribution of sequence variations between AH and normal populations. This is the first description of a specific gene defect associated with AH. PMID- 11932269 TI - Using baseline and recombinant human TSH-stimulated Tg measurements to manage thyroid cancer without diagnostic (131)I scanning. PMID- 11932270 TI - Is diagnostic iodine-131 scanning with recombinant human TSH useful in the follow up of differentiated thyroid cancer after thyroid ablation? AB - Measuring serum Tg and performing a diagnostic whole body scan (DxWBS) has become the standard for follow-up of patients with differentiated thyroid carcinoma. The primary aim of this study was to determine whether recombinant human TSH (rhTSH) stimulated Tg alone is sufficiently sensitive to identify residual cancer in patients with no clinical evidence of disease and undetectable or very low serum Tg levels during thyroid hormone (TH) therapy. A secondary aim was to investigate the frequency of tumor in such patients. One hundred and seven consecutive patients, aged 10.9-85.3 yr (median, 36.3), at the time of initial surgery who had Tg levels on TH therapy that were undetectable (95% < or =0.5 ng/ml) or low (4% 0.6 ng/ml, 1% 1.0 ng/ml) and who underwent rhTSH-stimulated testing 10 months to 35 yr (median, 3.5 yr) after initial thyroidectomy and (131)I ablation were retrospectively studied. Many (50%) were at high risk of tumor recurrence, and 5 had distant metastases during the course of their disease. In response to rhTSH, Tg ranged from 0.5 or less to 17.9 ng/ml, remaining at 0.5 ng/ml or less in 68 (64%) patients and increasing to levels between 0.6 and 2 ng/ml in 19 (18%) others and to levels higher than 2 ng/ml in 20 (19%) patients. Eleven patients (10%), all of whom had rhTSH-stimulated serum Tg levels above 2 ng/ml, were found to have persistent tumor in lung (4 patients), lymph nodes (5 patients, 3 with cervical central compartment, 1 bilateral cervical, and 1 with mediastinal nodes) identified by fine needle cytology, surgical pathology, posttherapy whole body scans, or computed tomography and, in two patients, with high serum Tg values alone (4.6 and 7.0 ng/ml after rhTSH and, respectively, 28.5 and 70.6 ng/ml after TH withdrawal), although in neither could the tumor site be identified. Thirteen patients (12%) were treated with surgery or (131)I, and in some cases both, as a result of the rhTSH studies; 10 had tumor, 1 had residual uptake in the thyroid bed visible on rhTSH-stimulated diagnostic whole body scan (DxWBS), and 2 had high serum Tg levels, presumably originating from a tumor site that could not be identified. A patient's tumor status, even in retrospect, usually was not predictable on the basis of Tg during TH therapy or tumor node metastasis status: among patients found to have tumor after rhTSH, serum Tg during TH therapy was 0.5 ng/ml or less in 55% and 0.6 ng/ml in 36%, and tumor node metastasis status was T2N1 or less in 82%. In no case did the rhTSH-stimulated DxWBS show the site of persistent tumor. There were correlations between visible thyroid bed uptake on DxWBS and quantitated (131)I uptake (r(2) = 0.11; P = 0.001), between DxWBS and rhTSH-stimulated Tg (r(2) = 0.54; P = 0.001), and between rhTSH-stimulated Tg and (131)I uptake (r(2) = 0.66; P = 0.0001). There was no statistically significant difference (P = 0.4) in bed (131)I uptake in patients with rhTSH stimulated serum Tg levels of 0.5 ng/ml or less compared with that in subjects with higher rhTSH-Tg levels. An rhTSH-stimulated Tg level greater than 2 ng/ml had a sensitivity of 100%, a negative predictive value of 100%, and a false positive rate of 9%. The rhTSH Tg had a substantially better performance than the other studies; the false negative rates were 64% for Tg higher than 0.5 ng/ml on TH therapy, 73% for rhTSH-stimulated DxWBS showing uptake, and zero for an rhTSH stimulated Tg more than 2 ng/ml. In conclusion, of 107 patients who were clinically free of disease, 10% had persistent tumor (4 with pulmonary metastases and 5 with regional disease) that was only identified with an rhTSH-stimulated serum Tg level greater than 2 ng/ml. This study shows that tumor amenable to early therapy may be found when rhTSH-stimulated serum Tg rises above 2 ng/ml without performing a DxWBS, which merely provides data concerning the completeness of thyroid ablation, but not persistent tumor. An elevated rhTSH stimulated Tg greater than 2 ng/ml warrants further study. PMID- 11932271 TI - Diagnostic 131-iodine whole-body scan may be avoided in thyroid cancer patients who have undetectable stimulated serum Tg levels after initial treatment. AB - The follow-up of differentiated thyroid cancer after total thyroidectomy and thyroid ablation is commonly based on serum Tg determination and 131-iodine ((131)I) diagnostic whole-body scan (WBS) performed in the hypothyroid state, 6 12 months after thyroid ablation. Based on the greater sensitivity of Tg measurement, with respect to WBS, the diagnostic yield of diagnostic WBS has been questioned in patients who are off L-T(4) therapy and have undetectable Tg levels. The aim of the present retrospective study was to evaluate the diagnostic relevance of (131)I WBS performed after thyroid remnant ablation, in patients with undetectable serum Tg and off thyroid hormone therapy. The study included 315 of 662 consecutive patients (47.6%) treated in our department between 1980 and 1990, who, at the first control WBS after thyroid ablation, had undetectable serum Tg levels in the hypothyroid state. There were 54 men (17%) and 261 women (83%), with a mean age of 40.9 +/- 13.1 yr (range, 12-76), followed for a mean of 12 +/- 2.8 (range, 9-19) yr. The control WBS was negative in 225 (71.4%) patients and positive for persistent areas of thyroid bed uptake, frequently of very low significance, in 90 (28.6%). No local or distant metastases were discovered. At the last follow-up visit (1999-2000), 281 (89.2%) patients showed complete remission, with undetectable serum Tg off L-T(4) and negative WBS. Persistent thyroid bed uptake, with undetectable levels of Tg, was observed in 29 patients (9.2%) studied during L-T(4) withdrawal. Only 2 patients (0.6%) experienced local recurrence (lymph-node metastases) during their follow-up. In conclusion, our data suggest that the presence of undetectable levels of serum Tg off L-T(4) at the time of the first control WBS after initial treatment, is highly predictive of complete and persistent remission. With the exception of detecting persistent thyroid bed uptake in a minority of cases, the control WBS has never given information that could influence the following therapeutic strategy. On this basis, we propose that the diagnostic (131)I WBS may be avoided in patients with undetectable levels of Tg off L-T(4). These patients may be monitored with clinical examination, neck ultrasound, and serum Tg measurements on L-T(4). PMID- 11932272 TI - Effect of estrogen replacement plus low-dose alendronate treatment on bone density in surgically postmenopausal women with osteoporosis. AB - This prospective randomized, double-blind, placebo-controlled, clinical trial was performed to evaluate the effectiveness of estrogens plus low-dose alendronate on bone metabolism. A total of 150 surgically postmenopausal women with osteoporosis were randomized in three groups: group A, micronized E2 (2 mg/d) plus standard dose alendronate (10 mg/d); group B, micronized E2 plus low-dose alendronate (5 mg/d); and group C, micronized E2 plus placebo (one tablet per day). In all women, bone mineral density (BMD) and serum bone metabolism markers were assessed at admission and every 6 months for 2 yr. After 2 yr, BMD significantly increased compared with baseline in all groups. The percentage BMD change was significantly higher in groups A and B than in group C. The differences in BMD detected between groups A and B were not statistically significant. Since the 6-month follow-up and throughout the study, serum osteocalcin and bone alkaline phosphatase levels and urinary deoxypyridinoline and pyrilinks-D excretion were significantly reduced in all groups. Serum bone alkaline phosphatase levels significantly decreased in groups A and B, without difference between them, in comparison with group C. In conclusion, in surgically postmenopausal osteoporotic women treated with estrogen replacement, the addition of alendronate at a low dose of 5 mg daily induces a gain of bone mass not significantly different in comparison with that obtained using a standard dose of 10 mg daily. PMID- 11932274 TI - Clinical relevance of thyroid fluorodeoxyglucose-whole body positron emission tomography incidentaloma. AB - Fluorodeoxyglucose (FDG) whole body positron emission tomography (PET) scan is increasingly used in the diagnostic work-up or follow-up of patients. In these conditions, positive PET scans with unexpected hot spots within the thyroid region could be defined as thyroid FDG-PET incidentaloma (in analogy with unexpected sonographic thyroid nodules). We describe eight consecutive patients referred to the endocrine department because of thyroid "hot spots," incidentally detected by whole body FDG-PET scan (September 1999 to March 2001). Using ultrasound, fine needle aspiration cytology (FNAC), and histology reports, we tried to identify the pathology underlying thyroid FDG-PET incidentaloma. FNAC showed an indication for surgery in all patients. Surgery has been performed in 7 patients. Malignancy was correctly identified in five patients: two medullary thyroid carcinomas, one with lymph node invasion, and three papillary thyroid carcinomas with invasion through the thyroid capsule in two of the PTC cases. In two patients with a positive FDG-PET scan, FNAC pointed to follicular neoplasms, and final histology reports showed follicular adenoma. In the remaining patient, FNAC revealed a follicular lesion, but surgery has not yet been performed. In conclusion, a small series of consecutive thyroid FDG-PET incidentaloma cases is presented and suggests a high rate of clinically relevant malignancies. PMID- 11932273 TI - Differential effects of oral estrogen versus oral estrogen-androgen replacement therapy on body composition in postmenopausal women. AB - Menopause is associated with decreased lean body mass and increased fat due to aging and declining hormone secretion. Estrogens or estrogen-progestins have been used to alleviate vasomotor symptoms. However, estrogen-androgen (E/A) therapy is also used for vasomotor symptom relief and has been shown to increase lean body mass while decreasing fat mass. The objective of this 16-wk, double-blind, randomized, parallel group clinical trial was to compare esterified estrogen plus methyltestosterone (1.25 mg estrogen + 2.5 mg methyltestosterone/d; E/A group) vs. esterified estrogen alone (1.25 mg/d; E group) on body composition. Forty postmenopausal women (mean age, 57 yr) participated. Compared with estrogen treatment alone, women in the E/A group increased their total lean body mass and reduced their percentage fat for all body parts (P < 0.05). After E/A treatment, there were statistically significant increases in lean body mass by 1.232 kg [0.181 +/- 0.004, 0.81 +/- 0.057, and 0.24 +/- 0.009 kg in the upper body (P = 0.021), trunk (P = 0.001), and lower body (P = 0.047), respectively]. In the E group, the increase was 0.31 +/- 0.004, 0.021 +/- 0.03, and 0.056 +/- 0.05 kg in the upper body, trunk, and lower body, respectively. In the E/A group, body fat was reduced by 0.90 kg (P = 0.18 for the trunk only), and percentage body fat declined by 7.4% (P < or = 0.05 for all body parts). Lower body strength increased by 23.1 kg (51 lb) in the E/A group vs. only 11 kg (24.25 lb) in the E group (P = 0.002 between groups). A statistically significant increase in weight (2.7 +/- 5.1 vs. 0.1 +/- 4.6 lb; P < 0.05) was observed in the E/A group compared with the E group. When subjects were given self-reporting questionnaires, more improvement was noted in sexual functioning and quality of life in the E/A group when compared with patients receiving E alone. There were no noteworthy side effects. In conclusion, E/A replacement therapy can improve body composition, lower-body muscle strength, quality of life, and sexual functioning in postmenopausal women. PMID- 11932275 TI - Lack of impact of radioiodine therapy in tg-positive, diagnostic whole-body scan negative patients with follicular cell-derived thyroid cancer. AB - Several reports have suggested a benefit from radioactive iodine (RAI) therapy in Tg-positive, whole-body scan-negative patients with follicular cell-derived thyroid cancer, who were said to have high rates of visualization of uptake in metastases after therapeutic doses of RAI. We sought to evaluate the rate of visualization of RAI uptake in these patients and determine the effect of such therapy on tumor progression and Tg levels. We studied 24 consecutive patients who had been treated with high-dose RAI, four of whom had no evidence of metastasis or persistent cancer. Our results showed that four patients had some uptake in posttherapy scans: in the neck, lung, and mediastinal metastases in one patient, in the thyroid remnant in two, and in a possible neck microrecurrence in one. In 13 patients with macrometastases-tumors 1 cm or greater-tumors progressed and serum Tg increased; five have died of thyroid cancer. The disease remained stable in the seven patients with micrometastases. We concluded that in high-risk patients with follicular cell-derived thyroid cancer with high Tg levels and negative diagnostic whole-body scans, only a small number showed meaningful uptake after high doses of RAI. Therefore, widespread use of empiric RAI therapy for such patients who have a large tumor burden should not be encouraged. PMID- 11932276 TI - Flaxseed improves lipid profile without altering biomarkers of bone metabolism in postmenopausal women. AB - The risk of cardiovascular disease and osteoporosis drastically increases at the onset of menopause. Phytoestrogens have been suggested to inhibit bone loss and protect the cardiovascular system, in part by improving lipid profiles. The purpose of the present study was to examine the effects of flaxseed, a rich source of the phytoestrogens called lignans, on lipid metabolism and biomarkers of bone turnover in postmenopausal women. Postmenopausal women who were not on hormone replacement therapy were assigned to one of two treatment groups in a double-blind randomized study. Women were asked to consume 40 g of either ground flaxseed or wheat-based comparative control regimen daily for 3 months. In addition, all subjects received 1,000 mg calcium and 400 IU vitamin D daily. Flaxseed supplementation lowered (P < 0.05) both serum total cholesterol and non high-density lipoprotein cholesterol by 6%, whereas the comparative control regimen had no such effect. Flaxseed regimen reduced serum levels of both low density- and high-density-lipoprotein cholesterol by 4.7% and triglyceride by 12.8%, albeit not statistically significant. Serum apolipoprotein A-1 and apolipoprotein B concentrations were significantly (P < 0.005) reduced by 6 and 7.5%, respectively, by the flaxseed regimen. Markers of bone formation and resorption were not affected by either of the treatments. The findings of this study indicate that flaxseed supplementation improves lipid profiles but has no effect on biomarkers of bone metabolism in postmenopausal women. PMID- 11932277 TI - Lipoprotein profile in subclinical hypothyroidism: response to levothyroxine replacement, a randomized placebo-controlled study. AB - The relationship between subclinical hypothyroidism (SCH) and an atherogenic lipoprotein profile is still controversial. We measured lipoproteins in 49 SCH patients by comparison with 33 euthyroid controls. Total cholesterol (TC), triglyceride, high-density lipoprotein cholesterol, low-density lipoprotein cholesterol (LDLc), apolipoprotein A(1), apolipoprotein B, and lipoprotein (a) [Lp(a)] were measured after an overnight fast. Patients were randomly assigned to levothyroxine therapy or placebo and re-evaluated after 6 months of euthyroidism. SCH patients showed significantly higher TC (P < 0.01), LDLc (P = 0.01), and apolipoprotein B (P = 0.001) levels than controls, positively correlated with baseline TSH levels (P = 0.003, P = 0.01, and P = 0.03, respectively). Elevated Lp(a) levels were significantly more frequent in SCH (P < 0.05) and associated with familial diabetes mellitus and/or coronary heart disease (P < 0.01). Levothyroxine treatment resulted in a significant decrease of both TC and LDLc concentrations (P = 0.003), in direct proportion to the respective baseline values (P < 0.05 and P < 0.01, respectively), whereas no change in Lp(a) level was observed. No changes occurred in the placebo group. In conclusion, only serum LDLc levels are increased specifically and reversibly in association with SCH. Altered Lp(a) values reflect a genetic influence rather than a reduced thyroid hormone action. PMID- 11932278 TI - Bone loss at the femoral neck in premenopausal white women: effects of weight change and sex-hormone levels. AB - To investigate whether bone loss occurs in the premenopause, we measured the bone mineral content (BMC), bone mineral density (BMD), and bone area in the spine (L2 L4), femoral neck, and total hip, as well as the sex hormone levels of 130 healthy premenopausal white women (age, 31-50 yr) at least three times over 1-9 yr. We found an increase in all three bone measurements at the spine but no change in volumetric density. Neither could we detect any age-related changes in any of the three measurements in the total hip. In contrast, we detected a significant decrease in femoral neck BMD over time, due to a decrease in BMC and increase in bone area. Greater loss in femoral neck BMD was associated independently with weight loss and lower levels of estrone sulfate or E2. Separating the women into those with FSH spikes (>20 IU/liter) and women with consistently low FSH, we found the latter group had smaller decrease in BMD and that the decrease was due less to a decline in BMC and more to an increase in bone area. In summary, femoral neck BMD decreases in premenopausal women, particularly those with lower levels of estrogens resulting from slowing ovarian function despite regular menses. This decrease can be offset by more rapid weight gain. PMID- 11932279 TI - Dehydroepiandrosterone supplementation and bone turnover in middle-aged to elderly men. AB - In the present placebo-controlled, double-blind study, we assessed the effect of dehydroepiandrosterone (DHEA) supplementation (90 mg orally/d) on bone turnover in 43 healthy men, 56-80 yr old. Placebo or steroid was given for 6 months, followed by a 1-month washout period and then a further 6 months of the opposite agent. Serum samples were collected at baseline 3, 6, 7, and 13 months and assayed for procollagen peptide, bone-specific alkaline phosphatase, and osteocalcin, all markers of bone formation. Measurements were also made of serum cortisol, DHEA/DHEA-S, E2 and free and total T. First void, fasting urine was collected at baseline, 6, 7, and 13 months and assessed for deoxypyridinoline, a marker of bone resorption. Mean serum DHEA and DHEA-S levels in treated men were increased approximately 3-fold ( approximately 2.2 ng/ml to approximately 6 ng/ml) and 4.5-fold ( approximately 1000 ng/ml to approximately 4500 ng/ml), respectively, after 6 months and returned to baseline after washout. Similarly, circulating E2 concentrations were also increased 1.4-fold (from approximately 16 23 pg/ml; P < 0.001), a finding not observed with any other measured hormone. Bone marker levels remained remarkably constant at each sampling interval; procollagen peptide at approximately 8.0 ng/ml; bone-specific alkaline phosphatase at approximately 21.0 U/liter; deoxypyridinoline at approximately 4.5 nmol/mmol Cr. Osteocalcin showed a transient reduction from approximately 10.2- 6.2 ng/ml, P < 0.005 to P < 0.001, at 3 months, but this decline was observed in both treated and controls. Stratifying the marker levels by age or baseline DHEA/DHEA-S levels did not affect the findings. We conclude that oral DHEA does not affect bone turnover in middle-aged to elderly men when used for a 6-month period at doses targeted to restore circulating levels of the steroid to that seen in young adults. PMID- 11932280 TI - Effect of estrogen versus testosterone on circulating osteoprotegerin and other cytokine levels in normal elderly men. AB - Recent studies have shown that estrogen (E) likely plays a dominant role in inhibiting bone resorption in normal elderly men. Because both E and T inhibit osteoclast development and activity, stimulate osteoclast apoptosis, and inhibit osteoblast production of IL-6, it is unclear why T is less potent than E in inhibiting bone resorption in vivo. Osteoprotegerin (OPG) binds to and inactivates RANKL, the final mediator of osteoclastogenesis. In vitro, OPG production is stimulated by E, and preliminary data suggest that T has the opposite effect. Thus, we analyzed serum for OPG levels from a study in which 59 elderly men (mean age, 68 yr) were made acutely hypogonadal using a GnRH agonist and were also placed on an aromatase inhibitor to block conversion of androgens to estrogens. They were studied first under conditions of physiologic E and T replacement, and then randomized to no replacement, replacement with E alone, T alone, or both E and T. E alone resulted in an 18.6 +/- 7.9% (mean +/- SEM) increase in serum OPG levels (P < 0.05), whereas T alone tended to decrease OPG levels (by 10.0 +/- 8.5%; P < 0.05 compared with E alone). Using a two-factor ANOVA model, there was a highly significant T effect (P = 0.006) on decreasing serum OPG levels. Serum TNF-alpha, IL-6, and IL-6 soluble receptor levels increased significantly in the men who had both E and T withdrawn, and the increases in TNF-alpha and IL-6sR were absent in the men treated with either E or T. However, due to the variability in these cytokine measurements, the ANOVA models were not significant for E or T effects. Taken together, these data suggest that in vivo, T decreases OPG levels, whereas E tends to have the opposite effect. These differential effects of E vs. T on OPG production may explain, at least in part, why T has weaker effects than E on inhibiting bone resorption in vivo in humans. PMID- 11932281 TI - Metformin therapy in obese adolescents with polycystic ovary syndrome and impaired glucose tolerance: amelioration of exaggerated adrenal response to adrenocorticotropin with reduction of insulinemia/insulin resistance. AB - Functional adrenal hyperandrogenism occurs in women with polycystic ovary syndrome (PCOS). Insulin, similar to its ovarian effect, may impact the regulation of adrenal steroidogenesis by modulating the activity of P450c17alpha, the rate-limiting enzyme in androgen biosynthesis. We previously demonstrated that obese adolescents with PCOS are severely insulin resistant and are at heightened risk for impaired glucose tolerance and type 2 diabetes. In the present study we tested the hypothesis that metformin therapy in obese adolescents with PCOS will attenuate the adrenal steroidogenic response to ACTH, with reduction of insulin resistance/insulinemia. Fifteen adolescents with PCOS and impaired glucose tolerance received 3 months of metformin (850 mg, twice daily) therapy. Pre- and posttherapy they had oral glucose tolerance testing, ACTH stimulation test, a 3-h hyperinsulinemic (80 mU/m(2).min)-euglycemic clamp to assess insulin sensitivity and a hyperglycemic clamp to assess insulin secretion. After 3 months of metformin treatment, glucose intolerance improved, with eight subjects having normal glucose tolerance. Total and free T decreased [1.5 +/- 0.2 vs. 1.0 +/- 0.1 nmol/liter (P = 0.022) and 41.3 +/- 8.3 vs. 22.2 +/- 2.1 pmol/liter (P = 0.028), respectively]. Insulin-stimulated glucose disposal increased (21.5 +/- 2.2 vs. 25.0 +/- 2.2 micromol/kg.min; P = 0.041). Fasting insulin and oral glucose tolerance test insulin and glucose area under the curve decreased significantly. ACTH-stimulated increases in androstenedione, 17 hydroxyprogesterone, and 17-hydroxypregnenelone were lower after metformin treatment [2.8 +/- 0.4 vs. 1.7 +/- 0.3 nmol/liter (P = 0.014), 7.0 +/- 0.6 vs. 5.3 +/- 0.5 nmol/liter (P = 0.011), and 30.4 +/- 3.7 vs. 25.7 +/- 4.2 nmol/liter (P = 0.054)]. Fasting insulin correlated with the 17-hydroxypregnenelone response to ACTH stimulation (r = 0.52; P = 0.008). In summary, metformin treatment of obese adolescents with PCOS and impaired glucose tolerance is beneficial in improving glucose tolerance and insulin sensitivity, in lowering insulinemia, and in reducing elevated androgen levels. Moreover, metformin therapy is associated with attenuation of the adrenal steroidogenic response to ACTH. Metformin therapy was well tolerated. In conclusion, double blind, placebo-controlled studies will determine whether insulin-sensitizing therapy corrects not only ovarian hyperandrogenism but also functional adrenal hyperandrogenism in adolescents with PCOS. PMID- 11932282 TI - The biological variation of insulin resistance in polycystic ovarian syndrome. AB - Increased insulin resistance (IR) is a cardinal feature of overweight patients with polycystic ovarian syndrome (PCOS). However, there are no data on the variability of IR for subjects with PCOS. The biological variation of IR (homeostasis model assessment model) was assessed by measuring IR at 4-d intervals on 10 consecutive occasions in 12 overweight PCOS patients (median age, 28 yr; range, 18-31 yr) and 11 weight-matched control women having regular menses and without PCOS (median age, 30 yr; range, 19-33 yr). The distribution of IR was log Gaussian in PCOS and Gaussian distribution in the control group. The IR in PCOS subjects was significantly greater than in the controls [mean (range), 5.85 U (1-42.1) vs. 1.67 U (0.48-3.49); P = 0.001]. After accounting for analytical variation, the mean intraindividual variance was also substantially greater in PCOS patients than in controls (mean, 1.19 vs. 0.23). As a consequence, at any level of IR, a subsequent sample must rise by more than 322% or fall by more than 31% to be considered significantly different from the first. IR, measured using the homeostasis model assessment model, is significantly greater and more variable for overweight patients with PCOS. Therefore, this inherent variability needs to be accounted for in studies of IR in PCOS. PMID- 11932283 TI - First trimester insulin resistance and subsequent preeclampsia: a prospective study. AB - Insulin resistance is implicated in the pathogenesis of preeclampsia, but prospective data are limited. SHBG, a marker of insulin resistance among nonpregnant individuals, has not been studied in detail during pregnancy. We conducted a prospective, nested, case-control study to test the hypothesis that increased insulin resistance, marked by reduced first trimester SHBG levels, is associated with increased risk of subsequent preeclampsia. First trimester SHBG levels were measured in 45 nulliparous women who subsequently developed preeclampsia (blood pressure, > or =140/90 mm Hg; proteinuria, either > or =2+ by dipstick or > or =300 mg/24 h, after 20 wk gestation) and in 90 randomly selected normotensive nulliparous controls. Compared with controls, women who developed preeclampsia had significantly reduced first trimester SHBG levels (302 +/- 130 vs. 396 +/- 186 nmol/liter; P < 0.01). Every 100 nmol/liter increase in SHBG was associated with a 31% reduced risk of preeclampsia [odds ratio (OR), 0.69; 95% confidence interval (CI), 0.55, 0.88; P < 0.01]. After adjusting for covariates in a multiple logistic regression model, the association between first trimester SHBG and preeclampsia remained significant (per 100 nmol/liter increase; OR, 0.66; 95% CI, 0.47, 0.92; P = 0.01). When subjects were stratified by body mass index (lean: body mass index, < 25 kg/m(2); overweight: body mass index, > or =25 kg/m(2)), overweight women had lower SHBG levels than lean women (286 +/- 156 vs. 410 +/- 166 nmol/liter; P < 0.01), and within each stratum, women with preeclampsia had lower SHBG levels than their respective controls. In a multivariable analysis, the association between SHBG and preeclampsia strengthened among lean women, such that every 100 nmol/liter increase in serum SHBG was associated with a 55% reduction in the risk of preeclampsia (OR, 0.45; 95% CI, 0.27, 0.77; P < 0.01), whereas in overweight women, the association was mitigated (OR, 1.02; 95% CI, 0.62, 1.69; P = 0.9). We conclude that increased early pregnancy insulin resistance is independently associated with subsequent preeclampsia. First trimester SHBG levels may be a useful biomarker for preeclampsia, especially among lean women who otherwise would be perceived to be at low risk. PMID- 11932284 TI - Menstrual cycle effects on the neurohumoral and autonomic nervous systems regulating the cardiovascular system. AB - Gonadal hormones may affect homeostatic mechanisms regulating the cardiovascular system. We investigated this relationship at five different crucial hormonal time points along the menstrual cycle. Eight eumenorrheic healthy subjects underwent a battery of autonomic tests, hemodynamics, and volume-regulatory hormone measurements. Fluid-regulatory hormones, plasma renin activity, and aldosterone increased along the luteal phase (P = 0.003 and 0.02, respectively), whereas rest supine-corrected hematocrit declined in the course of the menstrual cycle (P = 0.001). Plasma norepinephrine decreased from 1.4 +/- 0.2 to 0.95 +/- 0.1 nmol/liter (P < 0.02) [early follicular (EF) to late follicular]. Thereafter, concentrations gradually returned to EF levels. Lf to Hf domain ratio (spectral analysis of electrocardiogram) showed a difference from that of norepinephrine. The cardiovagal baroreflex sensitivity increased significantly along the luteal phase (P = 0.04). The dose of isoproterenol required to increase heart rate (HR) 15 beats per minute was 0.19 +/- 0.04 microg during the EF time point, and it increased to 0.39 +/- 0.06 microg during the late luteal time point (P = 0.05). However, blood pressure, HR, and their responses to orthostatic stress remained unchanged. Fluctuations in the ovarian hormones along the menstrual cycle are associated with unchanged blood pressure and HR, despite the significant variations in the different homeostatic mechanisms regulating the cardiovascular system. PMID- 11932285 TI - In vivo modulation of plasma free fatty acids in patients with familial combined hyperlipidemia using lipid-lowering medication. AB - One of the best studied aspects of the insulin resistance syndrome in familial combined hyperlipidemia (FCHL) is impaired insulin-mediated suppression of FFA by diminished inhibition of hormone-sensitive lipase (HSL). In vitro experiments have shown that stimulation of HSL activity by catecholamines is decreased in FCHL. The aim of this study was to investigate HSL inhibition by insulin and stimulation by endogenous catecholamines in vivo in FCHL patients. Twelve FCHL subjects using lipid-lowering medication and 12 controls underwent a mental stress test after random ingestion of either 50 g glucose or placebo. After ingestion of glucose, insulin concentrations increased from 76.8 +/- 21.5 pM to a maximum of 520.2 +/- 118.4 pM (P < 0.01) in FCHL and from 38.0 +/- 5.0 to 221.7 +/- 25.1 pM (P < 0.01) in controls. The percent decreases in plasma FFA during the first hour after glucose ingestion were similar in FCHL and controls (67 +/- 5% vs. 72 +/- 3%, respectively), suggesting a comparable inhibition of HSL in both. During the placebo test, FFA increased similarly in FCHL (56 +/- 9%) and controls (57 +/- 19%). In contrast, FFA concentrations did not change during mental stress after ingestion of glucose (from 0.17 +/- 0.02 to 0.15 +/- 0.02 mmol/liter in FCHL and from 0.11 +/- 0.02 to 0.12 +/- 0.02 mmol/liter in controls). In conclusion, the present study provides in vivo evidence for intact insulin-mediated suppression of FFA in FCHL, although this inhibition of HSL was achieved by higher insulin levels, suggesting insulin resistance at the level of HSL. Secondly, the induction of HSL activity by endogenous catecholamines in vivo is not decreased in FCHL, in contrast to earlier in vitro findings. Finally, catecholamine-induced HSL activation can be inhibited by insulin in a similar manner in both FCHL and controls. PMID- 11932286 TI - Benefits of long-term GH therapy in Prader-Willi syndrome: a 4-year study. AB - Obesity, poor growth, and hypotonia in children with Prader-Willi syndrome (PWS) are accompanied by abnormal body composition resembling a GH-deficient state. Hypothalamic dysfunction in PWS includes decreased GH secretion, suggesting a possible therapeutic role for GH treatment. While short-term benefits of treatment with GH have been shown, whether these beneficial effects are dose dependent and persist or wane with prolonged therapy remains uncertain. Effects of 24 additional months of GH treatment at varying doses (0.3, 1.0, and 1.5 mg/m(2).d) on growth, body composition, strength and agility, pulmonary function, resting energy expenditure (REE), and fat utilization were assessed in 46 children with PWS, who had previously been treated with GH therapy (1 mg/m(2).d) for 12-24 months. Percent body fat, lean muscle mass, and bone mineral density (BMD) were measured by dual x-ray absorptiometry. Indirect calorimetry was used to determine REE and to calculate respiratory quotient. A modified Bruininks Oseretski test of physical performance evaluated strength and agility. During months 24-48 of GH therapy, continued beneficial effects on body composition (decrease in fat mass and increase in lean body mass), growth velocity, and REE occurred with GH therapy doses of 1.0 and 1.5 mg/m(2).d (P < 0.05), but not with 0.3 mg/m(2).d. BMD continued to improve at all doses of GH (P < 0.05). Prior improvements in strength and agility that occurred during the initial 24 months were sustained but did not improve further during the additional 24 months regardless of dose. Salutary and sustained GH-induced changes in growth, body composition, BMD, and physical function in children with PWS can be achieved with daily administration of GH doses > or =1 mg/m(2). Lower doses of GH, (0.3 mg/m(2).d) effective in improving body composition in GHD adults, do not appear to be effective in children with PWS at sustaining improvement in body composition. PMID- 11932287 TI - Changes in bone density and turnover explain the reductions in incidence of nonvertebral fractures that occur during treatment with antiresorptive agents. AB - Some, but not all, antiresorptive agents have been shown to reduce the risk of nonvertebral fractures. Agents that significantly reduced nonvertebral fracture risk also appear to produce larger mean increases in bone mineral density (BMD) and reductions in biochemical markers (BCM) of bone turnover, compared with other agents. To examine the extent to which increases in BMD and reductions in BCM during antiresorptive therapy are associated with reductions in risk of nonvertebral fractures, we performed a meta-analysis of all randomized, placebo controlled trials of antiresorptive agents conducted in postmenopausal women with osteoporosis (i.e. prior vertebral fracture or low BMD) with available relevant data. A total of 18 such trials with usable data were identified, including a total of 2,415 women with incident nonvertebral fractures over 69,369 women-years of follow-up. Poisson regression was used to estimate the association between changes in BMD or BCM during the first year and overall reductions in risk of nonvertebral fractures (vs. the placebo group) across all trials. Larger increases in BMD and larger reductions in BCM were significantly associated with greater reductions in nonvertebral fracture risk. For example, each 1% increase in spine BMD at 1 yr was associated with an 8% reduction in nonvertebral fracture risk (P = 0.02). Mean BMD changes at the hip were smaller than at the spine, but the predicted net effect on fracture risk was the same; an agent that increases spine BMD by 6% at 1 yr reduces nonvertebral fracture risk by about 39%, and an agent that increases hip BMD by 3% at 1 yr reduces nonvertebral fracture risk by about 46%. The results also predict that a 70% reduction in resorption BCM would reduce risk by 40%, and a 50% reduction in formation BCM would reduce risk by 44%. It appears that either BMD or BCM changes are able to explain the effect of treatment, because a separate variable for treatment was not independently significant in any models. These data demonstrate that larger increases in BMD at both the spine and hip and larger reductions in both formation and resorption BCM are associated with greater reductions in the risk of nonvertebral fractures. Antiresorptive agents that do not produce large increases in BMD or large reductions in BCM do not appear to and would not be expected to decrease the risk of nonvertebral fractures. PMID- 11932288 TI - Musculoskeletal effects of the recombinant human IGF-I/IGF binding protein-3 complex in osteoporotic patients with proximal femoral fracture: a double-blind, placebo-controlled pilot study. AB - The administration of recombinant human IGF-I complexed with its predominant binding protein IGF binding protein-3 (rhIGF-I/IGFBP-3) may allow the safe administration of higher doses of IGF-I than can be accomplished with rhIGF-I alone. The aim of this randomized, double-blind, placebo- controlled pilot study was to evaluate the short-term safety and musculoskeletal effects of rhIGF I/IGFBP-3 in older women (aged 65-90 yr) with recent hip fracture. Within 72 h after the event, 30 patients received continuous administration of either placebo (n = 10), 0.5 mg/kg.d rhIGF-I/IGFBP-3 (n = 9), or 1 mg/kg.d rhIGF-I/IGFBP-3 (n = 11). Treatment was administered by sc infusion through a portable mini-pump for a total of 8 wk after hip fracture surgery, with patient follow-up to 6 months after surgery. Efficacy evaluations included a contralateral hip bone density determination, markers of bone turnover (including serum osteocalcin and urinary excretion of N-telopeptide), grip strength, and tests of functional ability. During the administration of rhIGF-I/IGFBP-3, mean serum levels of IGF-I significantly (P < 0.001) increased from 83 ng/ml to 289 ng/ml (0.5 mg/kg.d) and 393 ng/ml (1 mg/kg.d), respectively. Both doses were well tolerated, and no hypoglycemia or other therapy-induced side effects were observed. After an initial loss of hip bone density after hip fracture surgery, patients treated with 1 mg/kg.d rhIGF-I/IGFBP-3 regained a substantial portion of their femoral bone mass. At 6 months postfracture (4 months after the 2-month infusion), they showed a statistically not significant decrease from baseline in hip bone density (-2.6%, P = 0.53). Placebo-treated patients, on the other hand, failed to regain lost bone: at 6 months postfracture, bone density in the placebo group had declined by 6.1% (P = 0.04). Additionally, in patients treated with 1.0 mg/kg.d rhIGF-I/IGFBP-3, grip strength had increased from baseline by 11.4% by the end of the study (P = 0.04) whereas patients on placebo lost 11.6% from baseline (P = 0.16). This increase in muscle strength in the high-dose group was associated with a positive effect on functional recovery. We conclude that a 2-month infusion of rhIGF-I/IGFBP-3 in patients with recent hip fracture is feasible, safe, and well tolerated. Analyzing the effects on bone mass, muscle strength, and functional ability, we observed beneficial trends. In the context of a small exploratory study, these findings should be interpreted with caution, but they support the need for future trials to further assess the therapeutic potential of rhIGF-I/IGFBP-3 in elderly subjects with osteoporosis. PMID- 11932289 TI - Human growth hormone replacement in adult hypopituitary patients: long-term effects on body composition and lipid status--3-year results from the HypoCCS Database. AB - The Hypopituitary Control and Complications Study is an international surveillance study evaluating efficacy and safety of GH therapy of adult GH deficient patients in clinical practice. The present report examined baseline data from 1,123 adult onset (AO) and 362 childhood onset (CO) patients, as well as efficacy in 242 patients who had completed 3 yr of GH treatment. At study entry, mean height, body mass index, waist to hip ratio, and lean body mass were significantly (P < 0.001 for each) lower in CO compared with AO patients. After 3 yr on GH, lean body mass was significantly increased in AO males and females and CO males but not CO females, whereas fat mass was significantly decreased in AO males only. Serum total cholesterol was decreased in females (-0.32 +/- 1.00 mmol/liter; P = 0.045) and males (-0.36 +/- 0.96 mmol/liter; P = 0.004). High density lipoprotein (HDL) cholesterol was increased for females (0.10 +/- 0.26 mmol/liter; P = 0.026) and males (0.10 +/- 0.34 mmol/liter; P = 0.022). The low density lipoprotein/HDL ratio was decreased in AO males (-0.93 +/- 2.00; P = 0.003), AO females (-0.65 +/- 0.74; P < 0.001), and CO females (-0.69 +/- 0.76; P = 0.038), but the decrease in CO males was not significant (-0.84 +/- 2.85; P = 0.273). In AO patients, lean body mass increase from baseline was greatest in the those younger than 40 yr old, less but still significant in the middle group (40 60 yr) and unchanged in older (>60 yr) patients; conversely, decreases in the low density lipoprotein/HDL ratio were small and not significant in the younger patients but greater and significant in the middle and older age groups. During the 3-yr treatment, 114 (7.7%) patients discontinued, including 9 (0.6%) for tumor recurrences, 9 (0.6%) for neoplasia, and 9 (0.6%) for side effects. Therefore, these observational data showed significant long-term efficacy of adult GH replacement therapy on body composition and lipid profiles and indicate that age is an important predictor of response. PMID- 11932290 TI - Hypogonadotropic hypogonadism and cerebellar ataxia: detailed phenotypic characterization of a large, extended kindred. AB - Although the co-occurrence of cerebellar ataxia and hypogonadism has been recognized for close to 100 yr, cases of Gordon Holmes syndrome are quite rare. This report describes the largest kindred characterized to date. The parents of the three affected siblings are first cousins, suggesting that the disease was inherited as an autosomal recessive trait. The siblings' initial evaluation was notable for low serum levels of sex steroids and gonadotropins (consistent with hypogonadotropic hypogonadism), progressive ataxia, and dementia. Extended treatment with physiological doses of pulsatile GnRH failed to stimulate a gonadotropin response. Brain imaging revealed volume loss in the cerebellum, with extensive abnormalities in the cerebral white matter. This unique family suggests that a common genetic mechanism is responsible for the syndrome of progressive hypogonadotropism and cerebellar ataxia. PMID- 11932291 TI - Delayed puberty: analysis of a large case series from an academic center. AB - Despite the clinical importance of delayed puberty, the understanding of this condition is hampered by the lack of studies evaluating etiologies and predisposing factors among large case series. We performed a retrospective study of clinical and laboratory data from adolescents (< or =18 yr of age) with delayed puberty who had been seen in our clinic between 1/96 and 7/99 (n = 232 subjects; 158 males and 74 females). Family histories of pubertal timing among primary relatives were classified as negative, having at least a tendency to pubertal delay (development > or =1 SD beyond the mean), or diagnostic of delay (development > or =2 SD beyond the mean). The most common cause of delayed puberty was constitutional delay of growth and maturation (CD), which affected 53% of the subjects (63% of males and 30% of females). The remaining subjects could be divided into four categories: those with an underlying condition associated with delayed, but spontaneous, pubertal development [functional hypogonadotropic hypogonadism (FHH)], 19% of subjects; those with permanent hypogonadotropic hypogonadism, 12% of subjects; those with permanent hypergonadotropic hypogonadism, 13% of subjects; and those without clearly classified disorders, 3% of subjects. Like CD, FHH was male predominant, whereas the other categories either affected both genders equally or were predominantly female. In total, 50 different etiologies led to pubertal delay within our case series. Data permitted classification of family histories of pubertal timing among primary relatives in 95 of 122 of the CD and in 25 of 45 of the FHH cases. Analysis revealed at least a tendency to pubertal delay in 77% of the CD and in 64% of the FHH families and a diagnosis of delay in 38% of the CD and 44% of the FHH families. Both parents contributed to the positive family histories. The rates of positive family histories among the CD and FHH groups were approximately twice those seen among the other subjects in our case series. Among all subjects, those with FHH had the most marked growth delay, and girls had the greater bone age delay. Among the boys and at comparable chronological ages, CD and FHH were characterized by greater delays in pubic hair development and bone age than in the other diagnostic groups. Although CD is typically associated with leanness, 22% of our subjects had a BMI SD score at the 85th percentile or above for chronological age. These overweight subjects differed from the rest of the CD group: bone age was less delayed, and height was less affected. Finally, our analysis suggested a possible association between attention deficit disorder with or without hyperactivity and pubertal delay in our CD and FHH subjects. Our study provides valuable data regarding the variety and frequency of diagnoses that lead to delayed puberty. The results underscore the importance of performing a thorough evaluation and family history in adolescents with delayed puberty. Moreover, the data from our case series provide clues for unraveling the mechanism(s) of idiopathic pubertal delay and lead to the hypothesis that the pubertal delay seen among some subjects with FHH and CD may stem in part from similar underlying physiology. PMID- 11932292 TI - The cerebrospinal fluid/serum leptin ratio during pharmacological therapy for obesity. AB - The aim of the present study was to evaluate the cerebrospinal fluid (CSF)/serum leptin ratio during pharmacological therapy for obesity with centrally and peripherally acting drugs. Thirty-one obese women (mean age, 32.3 +/- 10 yr; body mass index, 38.2 +/- 5.2 kg/m(2); body fat, 43.3 +/- 5.4%) were studied before and 2 months after a weight loss program consisting of a balanced diet (1200 kcal/d) plus drug therapy. The patients were randomly assigned into three study groups: group I, fenproporex 25 mg/d (n = 10); group II, sibutramine 10 mg/d (n = 10); and group III, orlistat 120 mg tid (n = 11). Body fat, measured by dual energy x-ray absorptiometry, and serum and CSF concentrations of leptin were examined at baseline and 2 months after therapy. At baseline, clinical and biochemical characteristics of the groups were similar. All of the women lost weight, approximately 7.0% of their initial body weight, and the reduction was not different among the groups. Serum leptin fell significantly after 2 months in all groups, and the decline was proportional to the reduction in body fat, because leptin levels adjusted for body fat did not change after treatment. CSF leptin levels showed a significant decrease after 2 months in all groups, and this decline was higher on group III compared with group I (P = 0.006). After therapy, the CSF/serum leptin ratio did not change in group I (1.57 +/- 0.3 to 1.72 +/- 0.62%) and group II (1.78 +/- 1.01 to 1.69 +/- 1.27%), whereas it declined significantly in group III (1.65 +/- 0.43 to 1.09 +/- 0.47%; P < 0.01), corresponding to a decrease of 33.3 +/- 22.5% for the CSF/serum leptin ratio. The percentage change in group III was significantly different from the positive variation on group I (11.9 +/- 42.1%; P = 0.006) and close to the statistical significance compared with the negative variation seen in group II (-7.6 +/- 27.8%; P = 0.06). Our results showed that the CSF/serum leptin ratio decreased after weight loss in obese women treated during 2 months with orlistat, whereas this ratio did not change in this period of time in obese women treated with fenproporex and sibutramine. PMID- 11932293 TI - Ocular metastases secondary to carcinoid tumors: the utility of imaging with [(123)I]meta-iodobenzylguanidine and [(111)In]DTPA pentetreotide. AB - Ocular metastases from carcinoid tumors are considered rare. They can be the primary presentation of a carcinoid tumor or develop during the course of the disease. The extent of distant metastases from carcinoid tumors correlates with poor prognosis and survival; early detection of metastasis may change the overall management. Radiopharmaceutical-labeled imaging techniques have been widely applied for the detection and localization of such lesions based on isotope uptake by neuroendocrine tumors. Routine application of these imaging modalities may reveal previously unsuspected lesions and may also be used to help stage the disease and to identify patients who may be treated with radiopharmaceuticals. Of 40 patients with carcinoid tumors reviewed in our department since we started routine scanning, we identified 6 (15%) who demonstrated ocular metastases: 5 with obvious lesions and 1 with presumed metastasis according to the results of nucleotide scanning. All 6 were negative on screening with [(123)I]meta iodobenzylguanidine, whereas 3 of 4 who were screened with [(111)In]octreotide showed positive uptake. All patients responded well to radiotherapy and chemotherapy and did not require surgical treatment. The orbit and its contents appear to be a common site for carcinoid metastasis, and radiopharmaceutical imaging with labeled octreotide is useful in identifying many of these lesions. PMID- 11932294 TI - Combined treatment with testosterone (T) and ethinylestradiol (EE2) in constitutionally tall boys: is treatment with T plus EE2 more effective in reducing final height in tall boys than T alone? AB - Estrogens have been shown to rapidly inhibit longitudinal growth in tall boys. To antagonize the initial growth accelerating effect of T, 41 boys with an initial height prediction in excess of 203 cm were treated prospectively with either T enanthate (TE) 250 mg/wk im in combination with ethinylestradiol (EE2) 0.1 mg/d taken orally for the first 5.8 +/- 0.47 wk (mean +/- SE) of treatment (group 1, n = 20) or with TE alone (group 2, n = 21). Patients were randomized to one or the other treatment regimen. Mean (+/-SE) predicted adult height was 206.8 +/- 0.7 cm in group 1 and 206.4 +/- 0.8 cm in group 2. Total duration of treatment was 16.1 +/- 0.8 months and 14.0 +/- 1.2 months in group 1 and 2, respectively (NS). EE2 induced side effects in group 1 (gynecomastia) were limited and fully reversible. No negative long-term sequelae were found at final height with respect to hypothalamic-pituitary-gonadal axis activity and to testis volumes. Although there was a tendency to a lower initial growth velocity measured by knemometry in group 1 (0.30 +/- 0.05 vs. 0.38 +/- 0.05 mm/wk, NS), final height did not differ in both study groups (195.0 +/- 0.8 cm in group 1, 194.6 +/- 0.8 cm in group 2). Similarly, height reduction (initial predicted adult height minus final height) was not significantly different between the two groups (12.0 +/- 0.9 cm in group 1, 11.7 +/- 0.9 cm in group 2). In conclusion, the addition of EE2 during the initial treatment phase to high-dose T in tall boys has no significant effect on height reduction. The results of this clinical trial suggest that the initial growth inhibiting effect of EE2 on the epiphyseal growth plates is overridden by the long-term administration of high dose TE. PMID- 11932295 TI - Optimal response criteria for the human CRH test in the differential diagnosis of ACTH-dependent Cushing's syndrome. AB - The CRH test is in widespread use for the differential diagnosis of ACTH dependent Cushing's syndrome (CS). Despite the greater availability worldwide of human-sequence CRH (hCRH), there are no large series reporting the response criteria that best discriminate between Cushing's disease (CD) and the ectopic ACTH syndrome (EC) when using hCRH, rather than ovine-sequence CRH. We have, therefore, analyzed retrospectively the serum cortisol and plasma ACTH responses to hCRH in patients with ACTH-dependent CS, to develop response criteria that best discriminate between CD and EC. One hundred fifteen consecutive patients with proven ACTH-dependent CS were studied: 101 with CD (78 females; mean age, 40 yr; range, 10-73) and 14 with EC (7 females; mean age, 46 yr; range, 32-69). The response to hCRH was also studied in 30 normal volunteers (NVs; mean age, 29 yr; range, 20-44) with no clinical evidence of CS, and the results were compared. Following basal sampling at -15 and 0 min, hCRH (100 microg iv) was administered via an indwelling forearm cannula at 0900 h and serum cortisol and ACTH were measured at 15-min intervals for 2 h. The mean basal, peak, incremental, and percentage change in the serum cortisol and ACTH at all time points, and combination of time points, were calculated and analyzed to establish the best criteria to discriminate between CD and EC, and also between CD and NVs. The mean serum cortisol concentration in samples obtained at 15 and 30 min after CRH increased by at least 14% above the mean basal in 85 of 100 patients with CD, but in none with EC, giving a sensitivity of 85% at a specificity set at 100%. In contrast, the best plasma ACTH response of a rise of 105%, calculated from the maximal rise, gave only 70% sensitivity at 100% specificity. In the NVs, the maximum cortisol at the mean 15+30 min time point was 615 nmol/liter. Using the 15 and 30 min time points as the reference point, 71 of 100 patients with CD had a rise of serum cortisol greater than 14% and also showed an absolute cortisol level more than 615 nmol/liter. Serum cortisol responses to hCRH can be used to suggest the diagnosis of CD in the majority of patients with this condition, but it should only be used in conjunction with other biochemical and imaging modalities in establishing this important diagnosis. The measurement of plasma ACTH was less helpful in making this distinction, although it may have additional value in excluding ACTH-independent causes of CS. Although we believe that bilateral inferior petrosal sinus sampling remains the single most useful test in discriminating CD from EC in patients with ACTH-dependent CS, hCRH offers rapid diagnostic information and is a useful adjunctive test in establishing the presence of a possible ectopic source. PMID- 11932296 TI - The desmopressin and combined CRH-desmopressin tests in the differential diagnosis of ACTH-dependent Cushing's syndrome: constraints imposed by the expression of V2 vasopressin receptors in tumors with ectopic ACTH secretion. AB - The role of desmopressin, alone or in combination with CRH, in the differential diagnosis between Cushing's disease (CD) and ectopic ACTH secretion (EAS) still remains uncertain. Based on existing data, the desmopressin test is regarded as an alternative to the CRH stimulation test and, when given in combination with CRH, it has been suggested to completely discriminate between patients with CD and EAS. However, assessment of these tests has been limited in only a small number of patients with EAS. Desmopressin is a relatively specific V2 vasopressin receptor (V2R) agonist. Although expression of V3 vasopressin receptor (V3R) is common in tumors with EAS, the expression of V2R has not been extensively investigated. In the present study, we report our findings of the desmopressin and the combined CRH-desmopressin test in a series of patients with CD and EAS; also, the expression of V2R and V3R was investigated in tumors with EAS by a RT PCR method. We assessed a cohort of 31 patients with ACTH-dependent Cushing's syndrome, including 26 patients with CD and five cases with histologically confirmed EAS. To avoid bias of predetermined criteria, univariate curves of the receiver operating characteristics (ROC) were constructed by plotting the sensitivity against 1-specificity at each level of the percent cortisol (F) and ACTH responses to these tests. Following desmopressin administration there was an overlap of the percent F and ACTH responses among patients with CD and EAS, and the area under the ROC curve for both these responses was not significantly different than that occurring by chance. This was also true for the percent F response following the combined CRH-desmopressin test. However, the area under the ROC curve for the percent ACTH rise following the combined test was significantly different; the point of the ROC curve closest to 1 corresponded to a percent ACTH rise of 218% (88% sensitivity and 80% specificity). Expression of V2R and V3R mRNA was investigated in four of the five excised tumors with EAS and revealed the presence of the V2R in all, whereas the V3R mRNA was expressed in three of these cases. In conclusion, in this series the desmopressin test produced a significant overlap of responses between CD and patients with EAS and, therefore, is of limited value in the differential diagnosis of the ACTH dependent Cushing's syndrome. This is most probably due to the expression of the V2R in tumors with EAS. Moreover, following the combined CRH-desmopressin test only the ACTH but not the F responses were diagnostically useful, but still far from completely discriminating patients with CD and EAS. PMID- 11932297 TI - Supraphysiological doses of GH induce rapid changes in cardiac morphology and function. AB - GH is an agent widely used in sport to improve physical performance and has been proposed as adjunctive therapy in several clinical conditions. However, its short term effects on the normal human heart are poorly understood. Sixty young normal volunteers (30 males and 30 females) were enrolled in a multicenter, double blind, placebo-controlled study. All subjects were randomized to receive GH (0.03 or 0.06 mg/kg.d) or placebo. A complete Doppler-echocardiographic examination was performed at baseline and after 4 wk of treatment. Low-dose GH did not significantly affect echocardiographic parameters. In contrast, high-dose GH increased left ventricular mass index by 12% (P < 0.05). The type of growth response was concentric, because left ventricular wall thickness but not diameter increased, leading to a 10% increase of relative wall thickness. These structural changes were associated with functional changes, including a significant increase in cardiac index and a decrease in peripheral vascular resistance; diastolic function was not altered. Fractional shortening and systemic blood pressure were unchanged in the two treatment groups. In conclusion, administration of GH for 4 wk at doses that simulate GH abuse in sport caused a high cardiac output state associated with concentric left ventricular remodeling. PMID- 11932298 TI - Reduced 11beta-hydroxysteroid dehydrogenase type 2 activity is associated with decreased birth weight centile in pregnancies complicated by asthma. AB - Pregnancies complicated by asthma are associated with an increased risk of low birth weight. Currently, the mechanisms causing this outcome are unknown. To investigate whether impaired placental function may be a determinant, we measured placental 11beta-hydroxysteroid dehydrogenase type 2 (11beta-HSD2) activity, protein and mRNA, placental CRH mRNA, fetal cortisol, and fetal estriol concentrations at delivery. Asthmatic subjects were classified according to inhaled glucocorticoid intake during pregnancy and compared with a control nonasthmatic group. There was a 25% reduction in neonatal birth weight centile in asthmatic women who did not use inhaled glucocorticoid treatment. This was accompanied by significantly reduced placental 11beta-HSD2 activity, significantly increased fetal cortisol, and a trend toward increased placental CRH mRNA and reduced fetal estriol concentrations. The use of inhaled glucocorticoids for treatment was associated with birth weight centile, 11beta HSD2 activity, CRH mRNA, fetal cortisol, and estriol concentrations similar to control levels. There was a significant inverse correlation between fetal cortisol and fetal estriol concentrations across all groups. These studies demonstrate that inhaled glucocorticoid intake for the treatment of asthma is associated with improved placental function and fetal outcome, suggesting that inflammatory factors associated with asthma may be detrimental to fetal growth and development in these pregnancies. PMID- 11932299 TI - Relationship of calpain-10 genotype to phenotypic features of polycystic ovary syndrome. AB - Polycystic ovary syndrome (PCOS) is associated with an increased risk of impaired glucose tolerance and type 2 diabetes. Recent evidence suggests that variation in the gene encoding the cysteine protease calpain-10 influences susceptibility to type 2 diabetes. The present study was undertaken to determine whether variation in this gene is associated with quantitative traits pertinent to the pathogenesis of PCOS and diabetes. We studied 212 women with PCOS (124 white of European ancestry, 57 African-American, 13 Hispanic, 13 Asian-American, and 5 Middle Eastern). Each subject was genotyped for 3 DNA polymorphisms in the calpain-10 gene associated with type 2 diabetes (SNP-43, -19, and -63). The white and African-American subjects were examined for association of these polymorphisms with phenotypic features of PCOS and type 2 diabetes. There were not enough individuals in the other groups for similar genotype/phenotype analyses. Nineteen (9%) of the 212 women with PCOS were diabetic and were not included in the genotype/phenotype analyses. Twelve (63%) of these subjects were African American. Phenotypic traits in nondiabetic white probands did not differ whether analyzed for each individual SNP (SNP-43, -19, -63) or haplotype combination. Nor was there association of SNP-43, -19, or -63 with any of the phenotypic features of type 2 diabetes or PCOS in nondiabetic African-Americans. However, nondiabetic African-Americans with the 112/121-haplotype combination had significantly higher insulin levels, in response to an oral glucose challenge, as reflected in the area under the insulin curve (257,021 +/- 95,384 vs. 136,240 +/- 11,468 pmol/min; P = 0.03), compared with those with other haplotypes. This finding was particularly notable because the 112/121 subjects were less obese. The difference between groups in area under the insulin response curve remained significant (P = 0.002 by analysis of covariance) after adjustment for body mass index. In addition to its association with insulin levels in African-Americans, the 112/121 haplotype combination was associated with an approximate 2-fold increase in risk of PCOS in both African-Americans and whites. PMID- 11932300 TI - Familial medullary thyroid carcinoma: clinical variability and low aggressiveness associated with RET mutation at codon 804. AB - Sixty-one heterozygotes harboring the germline V804L mutation of the RET protooncogene were identified in five independent families. A total of 31 subjects underwent surgery. Histology identified C cell hyperplasia in 30 cases, isolated in 12 and associated with medullary thyroid carcinoma (MTC) in 18. Six patients with MTC had lymph node metastases. Among the 14 patients with basal detectable calcitonin (CT) level, 12 had MTC and 2 had isolated C cell hyperplasia. In most individuals carrying 804 RET mutation, C cell disease displayed late onset and an indolent course; a pentagastrin test was negative in the majority of heterozygotes during the first 2 decades and was positive in only half of them during the third and fourth decades of life. Interestingly, concomitant somatic M918T was detected in a 12-yr-old girl with MTC and was likely to be responsible for both the early clinical appearance and the aggressiveness of the disease. Our data show that in these gene carriers, surgery may be postponed to the fourth decade of life or until the pentagastrin stimulation test becomes positive. Indeed, our data should be confirmed on a larger series of V804L carriers, but may offer a balanced strategy to keep under control and prevent development of the full disease phenotype. PMID- 11932301 TI - Nongenomic cardiovascular effects of triiodothyronine in euthyroid male volunteers. AB - T(3) has been shown to exert cardiovascular effects. These effects have not yet been defined with regard to the mode of action (nongenomic vs. genomic) and with regard to an interaction with the adrenergic system in humans. To address these issues we conducted a randomized, double blind, 6-fold cross-over trial in 18 healthy male volunteers. After pretreatment with the beta-agonist dobutamine, the beta-blocking agent esmolol, or placebo (0.9% NaCl), 100 microg T(3) or placebo were injected. Primary target variables were systemic vascular resistance (SVR) and cardiac output (CO) within 45 min after injection of T(3) vs. placebo after placebo pretreatment. Sympatho-vagal balance was assessed by measurement of heart rate variability. T(3) caused a lower SVR and a higher CO than placebo (P < 0.001) after pretreatment with placebo. An increased low frequency (LF)/high frequency (HF) ratio (power in LF/power in HF band) after T(3) compared with placebo (P = 0.004) suggests an increase in sympathetic tone. After pretreatment with dobutamine, the effects of T(3) on SVR and CO were abolished, and the effect on LF/HF ratio was reversed. After pretreatment with esmolol, the effects on SVR and LF/HF ratio were reversed. Our data show, for the first time, nongenomic cardiovascular effects of T(3) in humans. PMID- 11932302 TI - Selenium supplementation in patients with autoimmune thyroiditis decreases thyroid peroxidase antibodies concentrations. AB - In areas with severe selenium deficiency there is a higher incidence of thyroiditis due to a decreased activity of selenium-dependent glutathione peroxidase activity within thyroid cells. Selenium-dependent enzymes also have several modifying effects on the immune system. Therefore, even mild selenium deficiency may contribute to the development and maintenance of autoimmune thyroid diseases. We performed a blinded, placebo-controlled, prospective study in female patients (n = 70; mean age, 47.5 +/- 0.7 yr) with autoimmune thyroiditis and thyroid peroxidase antibodies (TPOAb) and/or Tg antibodies (TgAb) above 350 IU/ml. The primary end point of the study was the change in TPOAb concentrations. Secondary end points were changes in TgAb, TSH, and free thyroid hormone levels as well as ultrasound pattern of the thyroid and quality of life estimation. Patients were randomized into 2 age- and antibody (TPOAb)-matched groups; 36 patients received 200 microg (2.53 micromol) sodium selenite/d, orally, for 3 months, and 34 patients received placebo. All patients were substituted with L-T(4) to maintain TSH within the normal range. TPOAb, TgAb, TSH, and free thyroid hormones were determined by commercial assays. The echogenicity of the thyroid was monitored with high resolution ultrasound. The mean TPOAb concentration decreased significantly to 63.6% (P = 0.013) in the selenium group vs. 88% (P = 0.95) in the placebo group. A subgroup analysis of those patients with TPOAb greater than 1200 IU/ml revealed a mean 40% reduction in the selenium-treated patients compared with a 10% increase in TPOAb in the placebo group. TgAb concentrations were lower in the placebo group at the beginning of the study and significantly further decreased (P = 0.018), but were unchanged in the selenium group. Nine patients in the selenium-treated group had completely normalized antibody concentrations, in contrast to two patients in the placebo group (by chi(2) test, P = 0.01). Ultrasound of the thyroid showed normalized echogenicity in these patients. The mean TSH, free T(4), and free T(3) levels were unchanged in both groups. We conclude that selenium substitution may improve the inflammatory activity in patients with autoimmune thyroiditis, especially in those with high activity. Whether this effect is specific for autoimmune thyroiditis or may also be effective in other endocrine autoimmune diseases has yet to be investigated. PMID- 11932303 TI - Cardiovascular risk factors in acromegaly before and after normalization of serum IGF-I levels with the GH antagonist pegvisomant. AB - Acromegaly is associated with premature cardiovascular mortality. GH replacement therapy decreases inflammatory markers of cardiovascular risk, but little is known about these markers in patients with acromegaly. The GH receptor antagonist, pegvisomant, reduces IGF-I levels in 98% of patients treated. We investigated the effects of GH receptor blockade on inflammatory and other cardiovascular risk markers in active acromegaly. Forty-eight patients with acromegaly and 47 age- and body mass index-matched controls were included. The study consisted of 3 parts: a cross-sectional study, a prospective randomized 12 wk placebo-controlled study, and a longitudinal open-label study of up to 18 months of pegvisomant treatment. After baseline evaluation, patients with acromegaly were randomized to placebo (n = 14), 10 mg (n = 12), 15 mg (n = 10), or 20 mg (n = 12) daily pegvisomant for 12 wk. Subsequently, all patients received at least 10 mg pegvisomant daily for up to 18 months, with dose adjustments to achieve a normal IGF-I level. Anthropometry, GH, IGF-I, and pegvisomant levels were measured monthly. C-reactive protein (CRP), IL-6, homocysteine, lipoprotein(a), glucose, insulin, triglycerides, total cholesterol, and high-density lipoprotein (HDL) and low-density lipoprotein (LDL) cholesterol were determined at baseline, 4 and 12 wk in the placebo-controlled study and at 3 month intervals (during which IGF-I levels were normal) in the longitudinal study. In the cross-sectional study, patients had lower CRP than did controls [median, 0.3 (range, 0.2-0.8) vs. 2.0 (0.6-3.7) mg/liter; P < 0.0001] and had higher insulin [78.6 (55.8-130.2) vs. 54.5 (36.6-77.5) pM, P = 0.0051]. IL-6, homocysteine, triglycerides, lipoprotein(a), LDL cholesterol and HDL cholesterol were not different between groups. In the placebo-controlled study, CRP increased in patients treated with 20 mg pegvisomant, compared with placebo (mean +/- SEM, 13.7 +/- 3.6 vs. 0.5 +/- 3.3 mg/liter; P = 0.010). There were no significant differences in IL-6, homocysteine, glucose, insulin, triglyceride, total cholesterol, LDL cholesterol and HDL cholesterol levels. In the longitudinal open label study (median duration, 15.6 months), CRP increased by 2.0 +/- 0.5 mg/liter (P = 0.0002). Total cholesterol and triglycerides increased (0.22 +/- 0.11 mM, P = 0.050; and 0.25 +/- 0.09 mM, P = 0.007, respectively), whereas lipoprotein(a) decreased (-70 +/- 33 mg/liter, P = 0.039). Glucose, insulin, homocysteine, HDL cholesterol, and IL-6 did not change. We conclude that patients with active acromegaly have lower CRP and higher insulin levels than healthy controls. Administration of pegvisomant increases CRP levels. We propose that GH secretory status is an important determinant of serum CRP levels, although additional studies are needed to determine the mechanism and significance of this finding. PMID- 11932304 TI - Characterization and semiquantitative analyses of pendrin expressed in normal and tumoral human thyroid tissues. AB - The gene mutated in Pendred syndrome (PDS), the PDS gene, is expressed in the inner ear, kidney, and thyroid. It encodes a membrane protein named pendrin that is endowed with the function of anion transporter or exchanger. It has been postulated that in the thyroid pendrin could participate in the transport of iodide from the cell to the lumen of follicles. We generated antipeptide antibodies directed against the C- terminal sequence of human pendrin 1) to characterize the protein expressed in the human thyroid, and 2) to analyze its expression level in relation to the functional activity of thyroid tissue. In denaturing conditions, a single molecular species of 110-115 kDa was identified in human thyroid membrane fractions. After treatment of thyroid membranes with N glycosidase F, pendrin had an apparent molecular mass of 85 kDa. Analyzed by ultracentrifugation on sucrose gradient in nondenaturing conditions, pendrin sedimented as a main 120- to 140-kDa component. Pendrin was assayed by semiquantitative Western blot in thyroid membrane fractions from 25 hyper- or hypofunctioning tumors and paired normal tissue samples. Pendrin was increased 2 fold in toxic adenomas, was not significantly altered in follicular adenoma, and was decreased, on the average, by 35% in papillary carcinomas compared with levels in paired normal tissue. The variations in the pendrin tissue content and PDS transcript levels, assayed by RT-PCR on duplicate samples of the same tumors, were similar. In conclusion, we show that pendrin expressed by the human thyroid gland is a mainly monomeric glycoprotein and that the level of expression of pendrin, although somewhat related, only moderately varied with the functional status of the thyroid tissue. PMID- 11932305 TI - Leptin and soluble leptin receptor levels in obese and weight-losing individuals. AB - To investigate soluble leptin receptor (sLR) in plasma, specific anti-sLR monoclonal antibodies were developed. Western blot analysis and size exclusion fractionation demonstrated sLR in plasma with a molecular mass of approximately 180,000. Next to this, the presence of sLR-leptin complexes in plasma was confirmed. Using the developed monoclonal antibodies, a specific sLR ELISA was developed, which measured in plasma both free and sLR bound to leptin. sLR appeared to inhibit leptin concentrations measured in four different leptin assays indicating that these assays primarily measure free leptin and underestimate the total leptin present in plasma. Furthermore, plasma levels of sLR and leptin were measured in 21 lean individuals and in 30 morbidly obese subjects before and 3, 6, and 12 months after gastric restrictive surgery. Preoperatively, leptin concentrations significantly correlated with body mass index (r = 0.796, P < 0.001). In contrast, sLR significantly inversely correlated with body mass index (r = -0.294, P < 0.05). In lean subjects, the molar ratio of free leptin to sLR was 1:1, whereas in morbidly obese subjects a ratio of 25:1 was found. After weight loss due to surgery, leptin levels rapidly decreased and sLR levels slowly increased to reach normal values at 12 months postoperatively. We conclude that sLR levels are significantly decreased, whereas leptin levels are significantly increased in morbidly obese subjects compared with lean individuals. PMID- 11932306 TI - Characterization and functional analysis of cAMP response element modulator protein and activating transcription factor 2 (ATF2) isoforms in the human myometrium during pregnancy and labor: identification of a novel ATF2 species with potent transactivation properties. AB - There is now extensive evidence to indicate that components of the cAMP signaling pathway are up-regulated in the human myometrium during pregnancy so as to potentiate the maintenance of uterine quiescence until term. In many tissue and cell types, increased signaling of the cAMP pathway results in profound changes in gene expression that are catalyzed via stimulation of PKA and activation of cAMP-dependent transcription factors that bind cAMP response elements (CREs) within the promoter regions of affected genes. In the myometrium, these CRE containing genes include beta2-adrenoceptor, cyclo-oxygenase 2, oxytocin receptor, and connexin-43. In preliminary investigations, we reported the differential expression of members of the cAMP bZIP protein family in the myometrium during pregnancy and labor. In this present study, we have now identified and functionally characterized these proteins with respect to myometrial gene expression. We report the identification of a 39,000 mol wt CRE response element modulator protein (CREM)tau2alpha protein having both transactivation and transrepressor properties whose expression is sequentially decreased in the myometrium during gestation and parturition. In contrast, expression of a myometrial 28,000 mol wt CREMalpha protein having only transrepressor actions progressively increased in the myometrium during pregnancy and labor. Similarly, we have isolated two ATF2 proteins of 60,000 and 28,000 mol wts, which represent full-length ATF2 and a novel small isoform of ATF2 that we have termed ATF2-small (ATF2-sm). These proteins are potent transactivators of gene expression and appear to be spatially expressed within the myometrium of the upper and lower uterine regions. The identification and functional characterization of these basic region/leucine zipper proteins in the myometrium may provide further insight into the molecular mechanisms regulating uterine activity during fetal maturation and parturition. PMID- 11932308 TI - Serum vascular endothelial growth factor levels are elevated in metastatic differentiated thyroid cancer but not increased by short-term TSH stimulation. AB - Solid tumor formation requires the development of a blood supply adequate to meet the metabolic demands of the enlarging tumor mass that cannot be sustained by simple diffusion. One principal stimulant to endothelial cell growth and migration, vascular endothelial growth factor (VEGF), is synthesized and secreted by thyroid cancer cells. Furthermore, VEGF overexpression is associated with an aggressive thyroid cancer phenotype in both animal models and clinical pathological studies. In other malignancies, elevated serum levels of VEGF often correlate with stage of disease and other poor prognostic clinical features. Therefore, we hypothesized that serum VEGF levels would be significantly higher in patients with persistent or recurrent thyroid cancer than in those cured of the disease. Because TSH stimulates both normal and neoplastic thyroid cells, we also proposed that serum VEGF would be further increased by TSH stimulation. Sixty-nine patients with either papillary or follicular thyroid cancer, status post total thyroidectomy, and prior radioactive iodine ablation, who had undergone routine recombinant human TSH (rhTSH, Thyrogen, Genzyme Transgenics Corp., Cambridge, MA) assisted whole-body radioactive iodine scanning, were included in this study. This cohort (mean age 53 +/- 16 yr, 51% female) included 21 patients with no evidence of disease and 48 patients with local or distant metastases. Stored serum samples obtained for standard Tg determinations before and 72 h following standard rhTSH stimulation were identified and assayed for VEGF 165 (R ?[amp ]? D Systems, Minneapolis, MN). Baseline serum VEGF levels obtained at a time of TSH suppression were significantly higher in patients with known metastatic disease than in those with no evidence of disease (416 +/- 62 pg/ml vs. 185 +/- 25 pg/ml, P = 0.001). Patients with distant metastases had baseline serum VEGF levels that did not differ significantly from patients with only cervical recurrences (455 +/- 90 pg/ml in distant metastases vs. 330 +/- 44 pg/ml for local cervical recurrences). Short-term TSH stimulation, although causing a significant rise in serum Tg, resulted in no significant increase in serum VEGF measured 72 h after rhTSH injection in either the patients with known metastatic disease (416 +/- 62 pg/ml baseline vs. 419 +/- 71 pg/ml after TSH stimulation) or in cured patients (185 +/- 25 pg/ml baseline vs. 191 +/- 33 pg/ml after TSH stimulation). Subgroup analysis revealed that patients with metastatic disease arising from well differentiated primary thyroid cancers had significantly higher serum VEGF levels than patients with metastatic disease arising from poorly differentiated thyroid cancer primaries (485 +/- 74 pg/ml vs. 167 +/- 32 pg/ml, P = 0.003 by ANOVA). Poorly differentiated metastatic thyroid cancers had serum VEGF levels indistinguishable from patients cured of disease (167 +/- 32 pg/ml vs. 186 +/- 25 pg/ml). In summary, serum VEGF is significantly elevated in patients with metastatic differentiated thyroid cancer but not in those with poorly differentiated thyroid cancer metastases. No measurable increase in serum VEGF levels can be detected 72 h after short-term TSH stimulation with rhTSH. We conclude that serum VEGF may serve as a clinical useful marker of residual differentiated thyroid cancer. PMID- 11932307 TI - Secreted frizzled related protein 1 is overexpressed in uterine leiomyomas, associated with a high estrogenic environment and unrelated to proliferative activity. AB - Secreted frizzled related protein 1 (sFRP1) is a modulator of Wnt signaling. Recently, aberrations of Wnt signaling were reported to be involved in the pathology of various human neoplasms. We investigated the expression and function of sFRP1 in uterine leiomyomas. Secreted FRP1 expression was increased in leiomyomas, compared with normal myometrium using Northern and Western blot analyses. Expression was strongest in the late follicular phase (high estrogenic milieu) of the menstrual cycle. Interestingly, expression was negligible in leiomyomas treated with GnRH agonist. Expression was also prominent in cells during E2 treatment, serum deprivation, and hypoxia. Moreover, induction of apoptosis by serum deprivation in a leiomyosarcoma cell line was enhanced by antisense inhibition of sFRP1. These results suggest that sFRP1 expression was associated with uterine leiomyomas, particularly under high estrogenic conditions. Secreted FRP1 expression was not associated with cell proliferation but rather occurred during cell protection against apoptosis in vitro. Strong sFRP1 expression under high estrogenic conditions seems to contribute to the development of uterine leiomyomas through the antiapoptotic effect of sFRP1, which appear to be independent of cell proliferation. PMID- 11932310 TI - GH increases extracellular volume by stimulating sodium reabsorption in the distal nephron and preventing pressure natriuresis. AB - Although sodium retention and volume expansion occur during GH administration, blood pressure is decreased or unchanged. The aim was to study the effect of short- and long-term GH replacement in adults on sodium balance, renal hemodynamics, and blood pressure. Ten adults with severe GH deficiency were included into a 7-d, randomized, placebo-controlled, cross-over trial followed by 12 months of open GH replacement. All measurements were performed under metabolic ward conditions. Extracellular water (ECW) was determined using multifrequency bioelectrical impedance analysis. Renal plasma flow and glomerular filtration rate were assessed using renal paraminohippurate and Cr(51) EDTA clearances, respectively. Renal tubular sodium reabsorption was assessed using lithium clearance. Plasma renin activity (PRA), plasma concentrations of angiotensin II, aldosterone, atrial natriuretic peptides and brain natriuretic peptides (BNP) and 24-h urinary norepinephrine excretion were measured. Seven days of GH treatment decreased urinary sodium excretion. Lithium clearance as a marker of proximal renal tubular sodium reabsorption was unaffected by GH treatment. ECW was increased after both short- and long-term treatment. This increase was inversely correlated to the decrease in diastolic blood pressure (r = -0.70, P = 0.02) between baseline and 12 months. Short-term treatment increased PRA and decreased BNP. The increase in PRA correlated with an increase in 24-h urinary norepinephrine excretion (r = 0.77, P < 0.01). Glomerular filtration rate and renal plasma flow did not change during treatment. The sodium- and water retaining effect of GH takes place in the distal nephron. The sustained increase in ECW in response to GH is associated with an unchanged or decreased blood pressure. This together with unchanged or decreased atrial natriuretic peptides and BNP may prevent pressure-induced escape of sodium. PMID- 11932311 TI - Intranasal administration of adrenocorticotropin-(1-24) stimulates adrenocortical hormone secretion. AB - To determine the efficiency of transmucosal absorption of ACTH, we measured serum cortisol, aldosterone, dehydroepiandrosterone (DHEA), and DHEA sulfate (DHEA-S) levels after intranasal (in) vs. iv administration of ACTH-(1-24) (250 microg) in 12 healthy adult men (mean age, 24.3 +/- 3.2 yr; range, 21-31 yr), who had received no prior medication and had no symptoms of rhinitis. Blood was collected at 0, 30, 60, 120, and 180 min after administration of ACTH-(1-24), and the levels of adrenocortical steroids were measured by specific RIAs. There were no side-effects associated with in or iv ACTH administration. After in administration, serum cortisol and aldosterone increased rapidly by 224.7 +/- 39.2% and 147.2 +/- 50.5%, respectively, peaking 30 min after ACTH-(1-24) administration, and decreasing to basal levels within 120 min. These increases in serum cortisol and aldosterone were lower than those obtained after iv administration. Thirty minutes after in or iv administration of ACTH-(1-24), DHEA increased by 49.1 +/- 27.2% and 81.6 +/- 17.1%, respectively, and remained elevated for 180 min. Serum DHEA-S levels did not change after in administration of ACTH-(1-24) and increased only slightly after iv injection. Adrenocortical steroid levels did not increase after in administration of saline. These data demonstrate that adrenocortical steroids are stimulated by in administration of ACTH-(1-24). We suggest that intranasal administration of ACTH offers both a diagnostic approach as an adrenal function test and a therapeutic approach as ACTH replacement therapy in patients with ACTH deficiency. The latter may be more physiological than glucocorticoid replacement. PMID- 11932312 TI - Demonstration of immunoglobulin G, A, and E autoantibodies to the human thyrotropin receptor using flow cytometry. AB - Human TSH receptor (TSHR) autoantibodies with biological activity result in thyroid dysfunction, but antibodies that simply bind do not. We have applied flow cytometry to the measurements of IgG, IgA, and IgE immunoreactivity to the TSHR in patients with Graves' disease (GD) and thyroid eye disease (TED) and in normal controls. CHO cells stably expressing the extracellular domain of the TSHR with a glycophosphatidylinositol anchor were produced and found to express approximately 4 times as many receptors, but of similar affinity, as JP09 in TSH binding studies. Substantial increases in median fluorescence and peak channel fluorescence were obtained by flow cytometry using TSHR monoclonal antibodies on the glycophosphatidylinositol cells. IgG autoantibodies were demonstrated in 55 of 65 untreated GD patients, 3 of 25 normal subjects, and 4 of 8 atypical TED sera (negative for TSHR autoantibodies with biological activity) by flow cytometry and correlated poorly with thyroid-stimulating antibodies. IgA antibodies were present in 1 of 12 normal, 1 of 7 treated GD with TED, and 3 of 8 atypical TED sera. IgE binding was observed in 1 of 12 normal, 2 of 8 treated GD without TED, 1 of 6 treated GD with TED, and 0 of 8 atypical TED sera. In conclusion, we have demonstrated autoantibodies that bind directly to the TSHR in the majority of GD patients and in 50% of patients with atypical TED and a small number of normal controls lacking TSHR antibodies that affect function. Although predominantly IgG lambda, TSHR autoantibodies of the IgA and IgE isotypes are also detectable. PMID- 11932314 TI - Early pregnancy levels of pregnancy-associated plasma protein a and the risk of intrauterine growth restriction, premature birth, preeclampsia, and stillbirth. AB - The risk of adverse perinatal outcome among 8839 women recruited to a multicenter, prospective cohort study was related to maternal circulating concentrations of trophoblast-derived proteins at 8-14 wk gestation. Women with a pregnancy-associated plasma protein A (PAPP-A) in the lowest fifth percentile at 8-14 wk gestation had an increased risk of intrauterine growth restriction [adjusted odds ratio, 2.9; 95% confidence interval (CI), 2.0-4.1], extremely premature delivery (adjusted odds ratio, 2.9; 95% CI, 1.6-5.5), moderately premature delivery (adjusted odds ratio, 2.4; 95% CI, 1.7-3.5), preeclampsia (adjusted odds ratio, 2.3; 95% CI, 1.6-3.3), and stillbirth (adjusted odds ratio, 3.6; 95% CI, 1.2-11.0). The strengths of the associations were similar when the test was performed before 13 wk gestation or between 13 and 14 wk gestation. In contrast, levels of free beta-human CG, another circulating protein synthesized by the syncytiotrophoblast, were not predictive of later outcome in multivariate analysis. PAPP-A has been identified as a protease specific for IGF binding proteins. We conclude that control of the IGF system in the first and early second trimester trophoblast may have a key role in determining subsequent pregnancy outcome. PMID- 11932315 TI - Fetal tissues are exposed to biologically relevant free thyroxine concentrations during early phases of development. AB - Maternal hypothyroxinemia in early pregnancy is often associated with irreversible effects on neuropsychomotor development. To evaluate fetal tissue exposure to maternal thyroid hormones up to midgestation, we measured total T(4) and free T(4) (FT(4)), T(3), rT(3), TSH, and possible binding proteins in first trimester coelomic and amniotic fluids and in amniotic fluid and fetal serum up to 17 wk. Samples were obtained before interruption of maternal-fetal connections. The concentrations in fetal compartments of T(4) and T(3) are more than 100-fold lower than those in maternal serum, and their biological relevance for fetal development might be questioned. We found, however, that in all fetal fluids the concentrations of T(4) available to developing tissues, namely FT(4), reach values that are at least one third of those biologically active in their euthyroid mothers. FT(4) levels in fetal fluids are determined by both their T(4) binding protein composition and the T(4) or FT(4) in maternal serum. The binding capacity is determined ontogenically, is independent of maternal thyroid status, and is far in excess of the T(4) in fetal fluids. Thus, the availability of FT(4) for embryonic and fetal tissues would decrease in hypothyroxinemic women, even if they were euthyroid. A decrease in the availability of FT(4), a major precursor of intracellular nuclear receptor-bound T(3), may result in adverse effects on the timely sequence of developmental events in the human fetus. These findings ought to influence our present approach to maternal hypothyroxinemia in early pregnancy regardless of whether TSH is increased or whether overt or subclinical hypothyroidism is detected. PMID- 11932316 TI - Mutations of the PDS gene, encoding pendrin, are associated with protein mislocalization and loss of iodide efflux: implications for thyroid dysfunction in Pendred syndrome. AB - Pendred syndrome (PDS) is an autosomal recessive disorder characterized by deafness and goiter. Phenotypic heterogeneity is observed in affected individuals, and thyroid dysfunction is particularly variable. The syndrome is caused by mutations in the PDS (SLC26A4) gene, encoding an anion transporter pendrin, which localizes to the apical membrane of thyroid follicular cells. PDS is thought to enable efflux iodide into the follicle lumen. More than 50 diseases causing mutations of PDS have been reported. Here we have investigated the effect of nine PDS missense mutations on pendrin localization and iodide transport with the view to understanding their functional impact. As demonstrated by transient expression of green fluorescent protein-tagged pendrin mutant constructs in mammalian cell lines, appropriate trafficking to the plasma membrane was observed for only two mutants. The remaining PDS mutants appear to be retained within the endoplasmic reticulum following transfection. Iodide efflux assays were performed using human embryonic kidney 293 cells transfected with mutant pendrin and cotransfected with sodium iodide transporter to provide a mechanism of iodide uptake. The results indicated loss of pendrin iodide transport for all mislocalizing mutations. However, PDS mutants are associated with variable thyroid dysfunction in affected subjects. We concluded that additional genetic and/or environmental factors influence the thyroid activity in Pendred syndrome. PMID- 11932318 TI - Altered GH elimination kinetics in type 1 diabetes mellitus can explain the elevation in circulating levels: bicompartmental approach. AB - Type 1 diabetes mellitus (DM 1) is associated with elevated circulating GH concentrations. Because these high GH levels could be explained either by an augmented pituitary secretion and/or delayed elimination clearance or distribution, we sought to evaluate GH pharmacokinetics to propose a model that better explains the elimination kinetics in patients with DM 1 and assess possible differences with normal volunteers that could justify elevation in GH circulating levels in these patients. A multicompartmental analysis was applied to serum GH concentrations measured at different times for 150 min in six patients with DM 1 and six age-, sex-, and body mass index-matched normal subjects after the administration of an iv bolus of recombinant human GH (200 microg), previous suppression of endogenous GH release with octreotide. The best fitting to the GH disappearance profiles was obtained with the biexponential equation in both groups. From it, we propose a bicompartmental model to explain GH kinetics in normal and diabetic patients. The mean transit time in both compartments and the mean residence time in patients with DM 1 were more than twice the values from control group. So in DM 1 elevated circulating GH concentrations are, at least partially, caused by a delayed GH plasmatic clearance. The DM 1 patients included in this study had a normal renal function; thus, our results agree with the hypothesis that DM 1 constitutes a GH insensitivity state because a reduced GH clearance by its receptor-mediated mechanism might explain the delayed GH elimination kinetics shown in patients with DM 1. However, the possibility of additional factors contributing to the slowed GH removal from circulation is not completely excluded. PMID- 11932319 TI - Association between AAAG repeat polymorphism in the P3 promoter of the human parathyroid hormone (PTH)/PTH-related peptide receptor gene and adult height, urinary pyridinoline excretion, and promoter activity. AB - The PTH/PTHrP receptor (PTHR1) plays an essential role in skeletal development and mediates many other functions of PTH and PTHrP. Human PTHR1 gene transcription is controlled by three promoters, P1-P3. The most proximal promoter, P3, is active in bone and osteoblast-like cell lines and accounts for the majority of renal transcripts in adults. We have identified a tetranucleotide repeat (AAAG)n polymorphism in the P3 promoter. In 214 unrelated Japanese, the repeat number (n) ranged from 3-8, with the AAAG5 allele being the most frequent (59%). In 55 unrelated Caucasians, n ranged from 5-7, and the frequency of the AAAG5 allele was 78%. The most frequent genotypes in a cohort of 85 young (18-20 yr) female Japanese were 5/5, 5/6, and 6/6. The 6/6 genotype was associated with greater height (5/5 vs. 6/6; P < 0.02) and lower urinary deoxypyridinoline and pyridinoline (P < 0.02), which are markers of bone resorption. The height of an additional 71 healthy female Japanese subjects, aged 14-17 yr, having genotype 5/5, 5/6, or 6/6 was also in the order of genotype 5/5 < 5/6 < 6/6 (5/5 vs. 6/6, P < 0.05). There was no significant difference in lumbar and femoral bone mineral density between genotypes. Likewise, there was no difference in circulating intact PTH levels between groups. The activity of P3 promoter-luciferase reporter constructs in transcription assays in 2 human osteoblast-like cell-lines varied according to repeat number, with AAAG6 being the least active. In conclusion, the P3 promoter (AAAG)n polymorphism is frequent in both Japanese and Caucasians and has potential as a linkage marker for the PTHR1 locus. In addition, it may influence the expression of the receptor in target tissues and have functional consequences on the developing skeleton. PMID- 11932320 TI - A comparison of the effects of pegvisomant and octreotide on glucose, insulin, gastrin, cholecystokinin, and pancreatic polypeptide responses to oral glucose and a standard mixed meal. AB - Standard medical therapy for patients with acromegaly includes somatostatin analogs. Owing to the widespread expression of somatostatin receptors, these may be associated with unwanted effects, such as altered glucose tolerance and impaired gut hormone release. Pegvisomant is a novel pegylated GH analog that competes with wild-type GH for GH-receptor binding sites but contains a position 120, amino acid substitution that prevents functional GH receptor dimerization, a known prerequisite for GH signal transduction and generation of IGF-I. We have studied the short-term effects of these two therapies (pegvisomant 20 mg/d for 7 d and octreotide 50 microg thrice daily for 7 d) on glucose tolerance and stimulated gut hormone release in six healthy male volunteers in an open-label, random-order, cross-over study. Subjects were assessed at baseline (oral glucose tolerance test and standard mixed meal) and on d 6 and 7 of each therapy with a minimum washout of 2 wk between treatments. Area under the curve and peak responses were analyzed using one-way repeated-measures ANOVA (on ranks where appropriate). Pegvisomant had no effect on glucose tolerance or stimulated gut hormone response during an oral glucose tolerance test and a standard meal. In contrast, octreotide significantly increased fasting plasma glucose, lowered fasting plasma insulin, and led to deterioration in glucose tolerance; three subjects developed impaired glucose tolerance and one diabetes mellitus by World Health Organization criteria. Octreotide significantly impaired stimulated release of cholecystokinin, gastrin, insulin, and pancreatic polypeptide. In conclusion, pegvisomant, unlike octreotide, is not associated with deterioration in glucose tolerance and impairment of stimulated gut hormone release in normal males. PMID- 11932322 TI - Cyclin D1 protein expression predicts metastatic behavior in thyroid papillary microcarcinomas but is not associated with gene amplification. AB - Overexpression of cyclin D1 occurs in several malignancies, often due to gene amplification, and this has been associated with aggressive tumor behavior, a higher incidence of lymph node metastases, and a poorer prognosis. The role of cyclin D1 in the pathogenesis of thyroid malignancy is unknown; however, cyclin D1 expression has been reported to occur in a proportion of well differentiated thyroid carcinomas. Micropapillary carcinomas of the thyroid are common incidental findings that almost always behave in an indolent manner and remain quiescent. However, rare microcarcinomas behave aggressively and metastasize early, giving rise to clinically significant disease. We hypothesized that cyclin D1 might play a role in the aggressive behavior of metastasizing papillary microcarcinomas. We reviewed the histopathology reports of 2,000 patients who underwent thyroid surgery at our institution between 1995-1999 and identified 22 patients who presented with gross regional metastases from a primary papillary microcarcinoma. These patients formed the index cohort for this analysis. As controls, we selected 34 patients with nonmetastasizing microcarcinomas. We studied these tumors for immunoreactivity to cyclin D1 on immunohistochemistry and analyzed 13 tumors that diffusely expressed cyclin D1 for gene amplification by differential PCR. Twenty of the 22 (90.9%) metastasizing papillary microcarcinomas expressed cyclin D1, compared with 3 of the 34 (8.8%) nonmetastasizing papillary microcarcinomas (P < 0.001). However, of the 13 tumors that showed diffuse immunoreactivity for cyclin D1 on immunohistochemistry, none showed amplification of the cyclin D1 gene on differential PCR. We conclude that cyclin D1 is significantly overexpressed in metastasizing papillary microcarcinomas of the thyroid. This is likely due to mechanisms other than gene amplification. Cyclin D1 immunohistochemistry may be a valuable tool in predicting metastatic potential in papillary microcarcinomas. PMID- 11932321 TI - Female pseudohermaphroditism caused by a novel homozygous missense mutation of the GR gene. AB - Familial glucocorticoid resistance is characterized by increased cortisol secretion without clinical evidence of hypercortisolism, but with manifestations of androgen and mineralocorticoid excess. This condition is mainly caused by mutations of the GR gene that cause inadequate transduction of the glucocorticoid signal in glucocorticoid target tissues. The clinical features of glucocorticoid resistance in females include hirsutism, acne, male pattern baldness, oligomenorrhea, and oligoanovulation. We describe here a new phenotype, female pseudohermaphroditism and severe hypokalemia, caused by a homozygous inactivating mutation of the GR gene. The proband was born with ambiguous genitalia from consanguineous parents and was mistreated as a 21-hydroxylase deficiency case since the age of 5 yr. She had very high levels of plasma ACTH (759 pg/ml or 167 pmol/liter) and high levels of cortisol (28-54 microg/dl or 772-1490 nmol/liter), androstenedione (5-14 ng/ml or 17-48 nmol/liter), T (174-235 ng/dl or 7-8 nmol/liter), and 17-hydroxyprogesterone (8-12 ng/ml or 24-36 nmol/liter). Her cortisol and 17-hydroxyprogesterone levels were not compatible with the diagnosis of classic congenital adrenal hyperplasia; furthermore, cortisol was not properly suppressed after dexamethasone administration (28 microg/d or 772 nmol/liter). Her laboratory evaluation indicated a diagnosis of glucocorticoid resistance. To investigate this puzzling clinical and biochemical picture, we analyzed both GR and CYP21 genes. Indeed, a homozygous T to C substitution at nucleotide 1844 in exon 5 of the GR gene was identified in the patient that caused a valine to alanine substitution at amino acid 571 in the ligand domain of the receptor. Her parents and an older sister were heterozygous for this mutation. A whole Epstein Barr virus-transformed cell dexamethasone-binding assay revealed that this Ala(571) mutant had a 6-fold reduction in binding affinity compared with the wild type receptor. In a functional assay using mouse mammary tumor virus promoter driven luciferase reporter gene, the mutant receptor displayed 10- to 50-fold less trans-activation activity than the wild-type receptor. In addition, a large heterozygous CYP21 conversion was identified in the patient and her father. In conclusion, we described the first case of female pseudohermaphroditism caused by a novel homozygous GR gene mutation. This phenotype indicates that pre- and postnatal virilization can occur in females with the glucocorticoid resistance syndrome. PMID- 11932323 TI - Overexpression of cyclin D1 and underexpression of p27 predict lymph node metastases in papillary thyroid carcinoma. AB - Lymph node metastasis in papillary thyroid carcinoma increases the morbidity of treatment and the risk of local regional relapse and may also affect cure rates and survival. Factors that predict lymph node metastasis are, however, unclear. We analyzed 125 patients with papillary thyroid carcinoma for factors that predict lymph node metastasis. On univariate analysis, age, extrathyroidal extension, tumor focality, overexpression of cyclin D1, and underexpression of p27 predicted lymph node metastasis, whereas patient gender and tumor size did not. On multivariate analysis, extrathyroidal extension, overexpression of cyclin D1, and underexpression of p27 proved to be strong independent predictors of lymph node metastasis. We suggest that immunohistochemistry for cyclin D1 and p27 will prove valuable in identifying papillary thyroid carcinomas with metastatic potential. PMID- 11932324 TI - Fibronectin, laminin, and collagen IV as modulators of cell behavior during adrenal gland development in the human fetus. AB - The specific development of the human fetal adrenal gland requires cell proliferation, migration, apoptosis, and zone-specific steroidogenic activity. The present work was designed to determine the physiological significance of the previously identified spatial distribution of extracellular matrix components in the fetal gland. Primary cultures of human fetal adrenal cells grown on collagen IV, laminin, or fibronectin revealed that cell morphology was affected by environmental cues. Matrices also modulated the profile of steroid secretion by the fetal cells. Collagen IV favored cortisol secretion after ACTH or angiotensin II stimulation and increased dehydroepiandrosterone production when the AT(2) receptor of angiotensin II was specifically stimulated. These effects were correlated by changes in the mRNA levels of 3beta-hydroxysteroid dehydrogenase and cytochrome P450C17. In contrast, fibronectin and laminin decreased cell responsiveness to ACTH in terms of cortisol secretion, but enhanced ACTH stimulated androgen secretion. Finally, extracellular matrices were able to orchestrate cell behavior. Collagen IV and laminin enhanced cell proliferation, and fibronectin increased cell death. This study is the first to demonstrate that the nature of extracellular matrix coordinates specific steroidogenic pathways and cell turnover in the developing human fetal adrenal gland. PMID- 11932325 TI - Gonadal determination and adrenal development are regulated by the orphan nuclear receptor steroidogenic factor-1, in a dose-dependent manner. AB - The orphan nuclear receptor steroidogenic factor-1 (SF-1, NR5A1) regulates the transcription of multiple genes involved in steroidogenesis, reproduction, and male sexual differentiation. A heterozygous loss-of-function SF-1 mutation (G35E) has been described in a patient with adrenal failure and complete 46XY sex reversal, indicating that haploinsufficiency of this factor is sufficient to cause a severe clinical phenotype. This mutation in the P-box region of the DNA binding domain markedly impairs SF-1 binding to most response elements. In an infant with a similar clinical phenotype, we identified an SF-1 mutation (R92Q) in a highly conserved residue of the A-box, a region that functions as a secondary DNA-binding domain. Strikingly, the affected infant was homozygous for the R92Q mutation, but three relatives (parents, sister) were phenotypically normal despite being heterozygous for the mutation. In functional assays, the R92Q mutant exhibited partial loss of DNA binding and transcriptional activity when compared with the G35E P-box change, consistent with its phenotypic expression only when transmitted as a homozygous trait. Taken together, these two naturally-occurring SF-1 mutations reveal the relative functional importance of the P-box and A-box regions for monomeric binding by nuclear receptors. In addition, these patients reveal the exquisite sensitivity of SF-1-dependent developmental pathways to gene dosage and function in humans. PMID- 11932327 TI - Response of serum macrophage migration inhibitory factor levels to stimulation or suppression of the hypothalamo-pituitary-adrenal axis in normal subjects and patients with Cushing's disease. AB - Macrophage migration inhibitory factor (MIF) is a proinflammatory pituitary and immune cell cytokine and a critical mediator of septic shock. It has been reported that MIF is secreted in parallel with ACTH from the pituitary in response to stress or inflammatory stimuli. MIF release from immune cells is also induced rather than inhibited by glucocorticoids. It has therefore been suggested that MIF may be a novel counterregulatory hormone of glucocorticoid action that acts both as a paracrine and endocrine modulator of host responses. We have measured circulating MIF levels, using a human MIF ELISA, in normal subjects and patients under numerous pathophysiological conditions. Serum MIF was measured in normal subjects who underwent stimulation of the hypothalamo-pituitary-adrenal axis with an insulin tolerance test (n = 8), a CRH-stimulation test (n = 5), a short synacthen test (n = 5), and following a low-dose dexamethasone suppression test (n = 6). We also sampled from a peripheral vein and both inferior petrosal sinuses before and after CRH stimulation in four patients with a histologically proven diagnosis of Cushing's disease. Immunostaining of the pituitary tumors for MIF was also performed. In normal subjects serum MIF levels did not rise in parallel with cortisol during the insulin tolerance or CRH test or after administration of synthetic ACTH. In all subjects cortisol levels became undetectable after the low-dose dexamethasone suppression test, and no consistent change was observed in serum MIF levels during the test. In patients with Cushing's disease, there was no basal central-to-peripheral gradient in MIF, and no consistent changes occurred in serum MIF levels in either the left or right inferior petrosal sinus after CRH stimulation; however, immunostaining of the surgically removed pituitary tumors from the same patients showed strong staining for both ACTH and MIF. These results show that in humans acute modulation of the hypothalamo-pituitary-adrenal axis does not significantly alter circulating MIF levels. In addition, ACTH-secreting pituitary tumors that express MIF do not release MIF either spontaneously or in response to CRH stimulation, and there is no gradient for MIF in the venous drainage of the pituitary. Our study suggests that the pituitary gland is not the major contributor to circulating MIF; an autocrine or paracrine role for pituitary-derived MIF is more likely. PMID- 11932328 TI - PPAR-gamma decreases endometrial stromal cell transcription and translation of RANTES in vitro. AB - An important step in the monthly turnover of the endometrial lining during the menstrual cycle is the cyclical recruitment and activation of inflammatory cells. Regulated Upon Activation Normal T Cell Expressed and Secreted (RANTES) has been shown to mediate inflammatory cell chemotaxis. This study investigated the effect of PPAR-gamma ligands upon transcription and translation of RANTES in human endometrial stromal cells. First, the expression of endogenous PPAR-gamma was confirmed in endometrial stromal cells. The human RANTES promoter was searched to identify likely PPAR response elements (PPREs), in which three putative sites were found. The effect of PPAR-gamma ligands upon RANTES promoter activity and protein production was analyzed. In cells transfected with RANTES promoter vectors containing 958 bp and 3 PPREs, the addition of PPAR-gamma ligands inhibited promoter activity by 60% (P < 0.01) and 48% (P < 0.02), respectively. Truncation of the gene promoter to delete all putative PPREs abrogated the ligand induced inhibition. Stromal cells showed a 40% decrease in RANTES protein secretion when treated with a PPAR-gamma ligand (P < 0.01). The use of PPAR-gamma ligands to reduce chemokine production and inflammation may be a productive strategy for future therapy of endometrial disorders, such as endometriosis. PMID- 11932330 TI - Characterization of an organic anion-transporting polypeptide (OATP-B) in human placenta. AB - Organic anion-transporting polypeptides (OATPs) are a family of multispecific carriers that mediate the sodium-independent transport of steroid hormone and conjugates, drugs, and numerous anionic endogenous substrates. We investigated whether members of the OATP gene family could mediate fetal-maternal transfer of anionic steroid conjugates in the human placenta. OATP-B (gene symbol SLC21A9) was isolated from a placenta cDNA library. An antiserum to OATP-B detected an 85 kDa protein in basal but not apical syncytiotrophoblast membranes. Immunohistochemistry of first-, second-, and third-trimester placenta showed staining in the cytotrophoblast membranes and at the basal surface of the syncytiotrophoblast. Trophoblasts that reacted with an antibody to Ki-67, a proliferation-associated antigen, expressed lower levels of OATP-B. OATP-B mRNA levels were measured in isolated trophoblasts under culture conditions that promoted syncytia formation. Real-time quantitative PCR estimated an 8-fold increase in OATP-B expression on differentiation to syncytia. The uptake of [(3)H]estrone-3-sulfate, a substrate for OATP-B, was measured in basal syncytiotrophoblast membrane vesicles. Transport was saturable and partially inhibited by pregnenolone sulfate, a progesterone precursor. Pregnenolone sulfate also partially inhibited OATP-B-mediated transport of estrone-3-sulfate in an oocyte expression system. These findings suggest a physiological role for OATP-B in the placental uptake of fetal-derived sulfated steroids. PMID- 11932329 TI - Vascular proliferation and vascular endothelial growth factor expression in the rhesus macaque endometrium. AB - The relationship between vascular endothelial growth factor (VEGF) expression and the pattern of vascular proliferation in the rhesus macaque endometrium has not been studied. In this report, we used in situ hybridization to evaluate VEGF, VEGF receptor type 1 and VEGF receptor type 2 mRNA expression during hormonally regulated menstrual cycles in ovariectomized macaques. Proliferating endothelial cells were identified by a double immunocytochemistry procedure that detected Ki 67 antigen and von Willebrand factor in the same endothelial cells. One and 2 d after progesterone withdrawal (premenstrual), VEGF mRNA was up-regulated in the glands and stroma of the superficial endometrial zones, a finding that supports our previous suggestion that VEGF may play a role in the menstrual induction cascade. During the postmenstrual repair phase, the healing surface epithelium showed a further, dramatic increase in expression of VEGF mRNA, accompanied by strong increases in signals for VEGF receptor types 1 and 2 in multiple profiles of small blood vessels immediately below the surface epithelium. This finding implicates VEGF in the early angiogenic processes associated with endometrial healing and regeneration. Vascular endothelial proliferation persisted throughout the cycle in the upper endometrial zones and showed a dramatic estrogen- dependent peak during the midproliferative phase. This proliferative peak coincided with a peak in VEGF expression in the endometrial stroma. Endothelial proliferation was also significantly correlated with the degree of stromal VEGF expression during the proliferative and secretory stages of the cycle. These results implicate VEGF of stromal origin in endometrial vascular proliferation. PMID- 11932331 TI - A longitudinal analysis of maternal serum insulin-like growth factor I (IGF-I) and total and nonphosphorylated IGF-binding protein-1 in human pregnancies complicated by intrauterine growth restriction. AB - In cord blood and late gestation maternal serum, IGF-I is positively correlated with birth weight, whereas IGF-binding protein-1 (IGFBP-1) is inversely correlated with birth weight. Our goal was to determine whether maternal serum or amniotic fluid concentrations of IGF-I, IGFBP-1, or nonphosphorylated IGFBP-1 (npIGFBP-1) in early gestation predict later fetal growth abnormalities. Maternal serum was collected prospectively across gestation (5-40 wk) from 749 pregnant subjects. Amniotic fluid was collected after amniocentesis during wk 15-26 from 207 subjects. We compared median serum concentrations of IGF-I, IGFBP-1, and npIGFBP-1 in 38 subjects who delivered growth-restricted infants with the control group of 236 subjects with normal weight infants for each gestational age grouping, wk 5-12, 13-23, and 24-34. In the control group median IGF-I concentrations were 14.8, 11, and 15.6 nmol/liter for wk 5-12, 13-23, and 24-34, respectively, compared with 13.7, 14.3, and 10.6 nmol/liter in the intrauterine growth restriction (IUGR) group. Median IGFBP-1 concentrations were 8.5, 30.4, and 24.4 nmol/liter, respectively, in controls, compared with 11.4, 28.6, and 25.5 nmol/liter in the IUGR group. Median npIGFBP-1 concentrations were 6.9, 22, and 17.4 nmol/liter, respectively, in controls, compared with 5.0, 32.1, and 24.2 nmol/liter in the IUGR group. In the control group the median amniotic fluid IGFBP-1 level was 13,160 nmol/liter, and the median npIGFBP-1 level was 15,970 nmol/liter; in the IUGR group these levels were 13,440 and 18,440 nmol/liter, respectively. No clinically useful differences were found between the IUGR and control groups. Our results do not support the use of maternal serum IGF-I or IGFBP-1 or amniotic fluid IGFBP-1 or npIGFBP-1 early in gestation to predict later fetal growth restriction. PMID- 11932332 TI - Possible activation of the renin-angiotensin system in the feto-placental unit in preeclampsia. AB - The purpose of this study was to elucidate the mechanisms underlying the regulation of feto-placental circulation mediated by the renin-angiotensin system under preeclamptic conditions. We measured angiotensin-converting enzyme (ACE) activity, protein expression, and mRNA expression in uncomplicated and preeclamptic placentas and examined the localization of ACE. In addition, ACE activity and mRNA expression in human umbilical venous endothelial cells (HUVECs) under hypoxic conditions were analyzed. ACE activity, protein expression, and mRNA expression in placental tissues from preeclampsia were all significantly higher than those from uncomplicated pregnancies. ACE activity in vessel fractions was extensively higher than that in trophoblast-rich or macrophage-rich fractions. Additionally, ACE activity in HUVECs was significantly higher than that in human arterial endothelial cells, and ACE mRNA was primarily localized to venous endothelial cells of stem villous in placentas. Furthermore, hypoxic condition induced both ACE activity and mRNA expression in HUVECs. These results suggested that venous endothelial cells within placental stem villous tissues and umbilicus play an important role in the regulation of the feto-placental renin angiotensin system, and in response to hypoxic conditions the feto-placental unit seemed to induce ACE activity in the placenta; such an effect would be likely to lead to regulation of the fetal circulation. PMID- 11932334 TI - Glial-derived neurotropic factor and RET gene expression in normal human anterior pituitary cell types and in pituitary tumors. AB - Glial-derived neurotropic factor (GDNF) signaling is mediated through a 2 component system consisting of the so-called GDNF receptor-alpha (GFRalpha1), which binds to GDNF. This complex activates the tyrosine kinase receptor RET. In this paper we demonstrate GDNF, GFRalpha1, and RET mRNA and protein expression in the human anterior pituitary gland. Double immunohistochemistry of anterior pituitary sections showed GDNF immunoreactivity in more than 95% of somatotrophs and to a lesser extent in corticotrophs (20%); it was almost absent in the remaining cell types. Also, although more than 95% of somatotrophs were stained for RET, no positive immunostaining could be detected in other cell types. Furthermore, we have looked for GDNF and RET in human pituitary adenomas of various hormonal phenotypes. Strong positive immunostaining was found for c-RET in all of the GH-secreting adenomas screened as well as in 50% of ACTH-producing adenomas. Positive immunostaining for GDNF was found in all of the GH-secreting adenomas and in 10% of the corticotropinomas. Lastly, we found strong positive immunostaining for GFRalpha1 in 90% of the somatotropinomas and 50% of the corticotropinomas as well as in 1 of 8 prolactinomas and 1 of 13 nonfunctioning adenomas. All of the remaining pituitary tumors screened were negative for RET, GDNF, and GFRalpha1. This study indicates that GDNF may well be acting in the regulation of somatotroph cell growth and/or cell function in the normal human anterior pituitary gland. The expression of RET in all of the somatotropinomas and in 50% of the ACTH-producing tumors implies that GDNF and RET could be involved in the pathogenesis of pituitary tumors. PMID- 11932335 TI - Incomplete modified fast in obese early pubertal girls leads to an increase in 24 hour growth hormone concentration and a lessening of the circadian pattern in leptin. AB - We studied nutrition and GH in eight obese girls, aged 6-11 yr. Blood was sampled every 15 min for 24 h. A 48-h diet providing 25% of assumed caloric needs was imposed, with repeat sampling during the last 24 h. Six nonfasting lean girls were also studied, and their mean GH was 3 times that of the obese girls in the fed state (P = 0.024). Dieting increased mean GH by 60% (P = 0.0028). There was no difference in pulse number for either group, but total secretion for lean girls was 3.9 times greater than that in obese girls during the fed state. With dieting, obese girls increased their total GH secretion by 60% (P = 0.010), but maintained lower total secretion, approximately 40% that of lean girls (P = 0.014). Mean leptin in obese girls in the fed state was 6.2 times greater than mean leptin in lean girls (P = 0.0001), with higher concentrations at night (P < 0.05) and lowering of total mean leptin while dieting. We conclude that in early pubertal obese girls, short-term caloric restriction partially reverses the low GH state that is characteristic of obesity. The change is concomitant with a decrease in leptin and a lessening of circadian differences. PMID- 11932336 TI - Genetic variability in insulin action inhibitor Ikkbeta (IKBKB) does not play a major role in the development of type 2 diabetes. AB - Recent evidence indicates that IkappaB kinase beta (Ikkbeta) may be a mediator of acquired forms of insulin-resistance. In this study, we examined whether genetic variability at the Ikkbeta locus (IKBKB) contributes to the development of genetic forms of early-onset type 2 diabetes transmitted with an autosomal dominant mode of inheritance. Linkage with four markers flanking the IKBKB gene was evaluated in 32 multigenerational families. Included in the study were 233 diabetic (mean age at Dx = 37 +/- 18) and 152 nondiabetic subjects. The overall LOD scores were negative (-54.9 and -46.2 on the centromeric and telomeric sides, respectively) indicating that variability in IKBKB was not a major determinant of diabetes in these families. Positive values, however, were observed for selected pedigrees. All 17 families for which linkage with the IKBKB locus could not be excluded were screened for sequence differences in the 22 exons and 1.6 kb of the 5' flanking region by dideoxyfingerprinting or direct sequencing. Polymorphisms were identified in the 5' flanking region (-1775del/insC and -1547T > A), exon 11 (c.1083A > G, L361L) and in intron 12 (IVS12+14t > a). However, no mutations segregating with diabetes could be found in these families. Furthermore, all four polymorphisms had similar allele frequencies in the 32 family probands, 171 individuals with common, later-onset type 2 diabetes, and 182 nondiabetic controls. We conclude that sequence differences in the IKBKB gene do not play a major role in either early-onset, autosomal dominant type 2 diabetes, or common forms with a later-onset. PMID- 11932337 TI - IL-15 regulation in human endometrial stromal cells. AB - A greater knowledge of IL-5 regulation within human endometrium is important in understanding key reproductive events and uterine Natural Killer cell function. In the present study, expression of IL-15 mRNA was shown to be up-regulated by both PGE(2) and IFN-gamma in cultures of human endometrial stromal cells (ESC). Release of IL-15 protein was also shown to be under the control of PGE(2) and IFN gamma using an enzyme-linked immunosorbent assay for IL-15. In addition, 8-Bromo cAMP was able to increase IL-15 release from ESCs (P < 0.005) implying the actions of PGE(2) may be via this second messenger. Addition of a progestin appeared to enhance these effects. Real-time quantitative PCR has demonstrated an up-regulation in IL-15 mRNA expression in the late secretory phase of the menstrual cycle (P < 0.005) and a progressive rise in IFN-gamma expression throughout the secretory phase and into first trimester decidua. These results suggest that IL-15 regulation in the human endometrium is complex and that hormonal control may be indirect. PMID- 11932338 TI - Insulin, unlike food intake, does not suppress ghrelin in human subjects. AB - Food intake suppresses plasma levels of the gastric peptide ghrelin in humans. We hypothesize that the food intake- suppression of ghrelin could be secondary to the plasma glucose and insulin changes after a meal. The aim of this study was to compare the effect of the administration of a combined pulse of glucose and insulin to the effect of one meal on plasma ghrelin in human subjects. A secondary aim was to study the effect of an oral glucose load on ghrelin levels. METHODS: Experiment 1 (n = 10) studied plasma glucose, insulin, leptin and ghrelin for 6 hours after a 790 kcal liquid meal. In Experiment 2 (n = 7), a subcutaneous pulse of insulin (Humalog 0.03U/kg) and an I.V. infusion of glucose were administered in order to mimic the plasma changes of glucose and insulin after a meal, and plasma ghrelin levels were monitored for 9 h. The OGTT data was used to study the effect of oral glucose on ghrelin. RESULTS: A mixed liquid meal decreased basal serum ghrelin by 26% at 40 minutes (p = 0.009). A 75 gr oral glucose load suppresses ghrelin by 28% at 30 minutes. Contrary to the meal effect, the parenteral administration of insulin and glucose did not suppress serum ghrelin. CONCLUSION: Unlike food intake, the administration of insulin and glucose does not suppress ghrelin levels. These data suggest that the suppressive effect of food intake or oral glucose on serum ghrelin is unlikely mediated by the changes of plasma insulin and glucose observed after the ingestion. PMID- 11932340 TI - Evidence for limiting ovarian tissue harvesting for the purpose of transplantation to women younger than 40 years of age. PMID- 11932341 TI - Analysis of the separate secretion of very low-density lipoprotein (VLDL)-1 and VLDL-2 by the liver will be a principal factor in resolving the proatherogenic lipoprotein profile in hypopituitarism. PMID- 11932342 TI - What treatment should be given to the cryopreserved ovarian tissue after thawing? PMID- 11932344 TI - Laboratory and clinical experience in 55 patients with macroprolactinemia identified by a simple polyethylene glycol precipitation method. PMID- 11932345 TI - Acromegaly and colorectal cancer: risk assessment should be based on population based studies. PMID- 11932346 TI - Hyperparathyroidism due to the so-called bone hunger syndrome in prostate cancer patients. PMID- 11932348 TI - SHOX: a geneticist's view. PMID- 11932352 TI - Endocrine-related resources from the National Institutes of Health. PMID- 11932353 TI - National Hormone & Peptide Program--NIDDK: recombinant hormones, hypothalamic peptides & other hormones, antisera, reagents, & hormone assay services available. PMID- 11932354 TI - Data handling in cancer clinical trials-how we can minimize potential biases. PMID- 11932355 TI - Methylation status and expression of human telomerase reverse transcriptase mRNA in relation to hypermethylation of the p16 gene in colorectal cancers as analyzed by bisulfite PCR-SSCP. AB - BACKGROUND: The expression level of human telomerase reverse transcriptase (hTERT) is correlated with telomerase activity and is expressed at high levels in malignant tumors. It is of interest whether expression of hTERT is regulated by methylation of the CpG island in the promoter of the hTERT gene. We examined hTERT expression and methylation status of the hTERT and other genes including p16. METHODS: We analyzed methylation status by bisulfite treatment and polymerase chain reaction with single-strand conformation polymorphism analysis (PCR-SSCP) and expression of the hTERT by RT-PCR, in 13 cancer cell lines, eight white blood cell samples and 24 colorectal cancer tissues. RESULTS: In the cancer cell lines, hTERT was expressed and the CpG island of the hTERT promoter was methylated. Most colorectal cancer tissues showed similar results. The promoter of hTERT was methylated in six cases, partially methylated in 17 cases and unmethylated in one case. All cases with methylation of hMLH1 or p16 also showed methylation of hTERT; however, some of the cases lacking p16 methylation also had hTERT methylation. CONCLUSION: Increased expression of hTERT is related to hypermethylation of hTERT in colorectal cancerous tissues as well as some cancer cell lines and disconcordant with hypermethylation of p16. PMID- 11932356 TI - Effect of a 3-hour interval between methotrexate and 5-fluorouracil in the treatment of metastatic colorectal cancer. AB - OBJECTIVE: This phase II study was designed to evaluate the efficacy and toxicity of 3-h interval sequential methotrexate (MTX) and 5-fluorouracil (5-FU) with leucovorin (LV) rescue in the treatment of patients with metastatic colorectal cancer. METHODS: Forty-two patients with histologically confirmed metastatic colorectal cancer and at least one two-dimensionally measurable lesion, aged 30 74 years old, with performance status < or =2 and no or one prior chemotherapy were selected. Patients received sequential MTX 100 mg/m2 by bolus injection and 5-FU 600 mg/m2 at 3 h followed by LV rescue initiated after 24 h (15 mg per body every 6 h for six doses). The treatment was repeated every week or every 2 weeks until disease progression. All patients were treated as out-patients unless complications arose. RESULTS: All 42 patients entered in this study were assessable both for response and toxicity. Fifteen patients achieved objective responses (one complete and 14 partial), for an overall response rate of 36% (95% CI: 11-51%). Response rates in pretreated patients and patients with naive chemotherapy were 27 and 42%, respectively. Sixteen patients had stable disease and 11 progressed with therapy. The median survival for all patients was 378 days. The hematological toxicity was mild with no grade 3/4 leukopenia. The major non-hematological toxicity was diarrhea (one grade 4, four grade 3). CONCLUSIONS: This 3-h interval sequential MTX and 5-FU with LV rescue is an active regimen in patients with metastatic colorectal cancer. The treatment showed mild toxicity and was administered on an out-patient basis. The present findings suggest that this regimen warrants further investigation in patients with metastatic colorectal cancer. PMID- 11932357 TI - Optimal timing of radical cystectomy for patients with invasive transitional cell carcinoma of the bladder. AB - OBJECTIVE: To determine whether the timing of radical cystectomy affects the survival of patients with invasive transitional cell carcinoma (TCC) of the bladder. METHODS: Between January 1985 and June 2000, 167 patients underwent radical cystectomy in our institution. Among them, 50 patients who did not receive any perioperative therapies, including chemotherapy and radiotherapy, were divided into two groups, viz. 28 patients who underwent radical cystectomy within 3 months after the primary diagnosis of invasive bladder cancer (group A) and 22 who underwent radical cystectomy more than 3 months after the primary diagnosis (group B), and we then compared several clinicopathological factors and the survival between these two groups. RESULTS: No significant difference was observed in the patients' clinicopathological characteristics except for age between these two groups. The median follow-up periods for groups A and B were 53 and 48 months, respectively. In groups A and B, an average of 1.2 (range 1-2) and 1.4 (range 1-3) transurethral resections (TURs) of bladder cancer were performed, respectively (p = 0.83). Twenty of 28 patients in group A underwent orthotopic neobladder replacement, whereas only four of 22 underwent neobladder creation (p = 0.001). The recurrence-free, cause-specific and overall survival rates in group A were significantly higher than those in group B (p < 0.05, p < 0.05 and p < 0.05, respectively). Final pathological analysis revealed that the distributions of pathological stage, tumor grade and lymph node metastasis were similar between groups A and B; however, the incidence of vascular involvement in group B was significantly higher than that in group A (p < 0.05), despite the lack of a significant difference in the incidence of vascular involvement in TUR specimens between these two groups. CONCLUSIONS: These findings suggest that radical cystectomy in the early disease process, especially before the occurrence of vascular involvement, may result in the improvement of survival of patients with invasive TCC of the bladder. PMID- 11932358 TI - The Influence of handling censored data on estimating progression-free survival in cancer clinical trials (JCOG9913-A). AB - BACKGROUND: Progression-free survival (PFS) is a common endpoint in cancer clinical trials. This study was undertaken to assess the impact of data errors and data handling on the statistical estimation of PFS. METHODS: Data from four trials conducted by the Japan Clinical Oncology Group were examined. Three types of data handling methods were defined: (1) data handling method A (METHOD-A), the collected event data are used as much as possible, (2) METHOD-C, only reliable data with firm evidence are used, and (3) METHOD-B is intermediate between METHOD A and METHOD-C. To assess the impact of each of the three methods, Kaplan-Meier survival curves, median PFS, proportion of PFS, log-rank p values and hazard ratios were estimated. RESULTS: In three trials that collected PFS data periodically, no remarkable differences in median PFS and the proportion of PFS were observed. In one trial with non-periodic data cleaning, however, the ratio of median PFS by METHOD-C to that by METHOD-B was 0.85, the maximum difference of proportion of PFS between METHOD-C and METHOD-B was 12.0% and the largest spread in PFS curves amongst the three methods was observed in this trial. In all trials, log-rank p values and hazard ratios for between arm comparisons did not differ between the three methods. CONCLUSIONS: Periodic data management can reduce errors in comparisons of PFS and is a critical requirement when using PFS as a major endpoint. Furthermore, proper data handling is essential in the estimation of patient benefit and caution is needed when making clinical decisions based on PFS. PMID- 11932360 TI - Redevelopment of small cell lung cancer after a long disease-free period: a case report. AB - Although it has been reported that the risk of second malignancies increases in long-term survivors of small cell lung cancer (SCLC) and late recurrence of SCLC is sometimes experienced, it seems uncommon. We recently experienced a case of redevelopment of SCLC after a long disease-free period. The case was considered to be second primary SCLC more than 10 years after the initial treatment. The necessity for lifelong follow-up in long-term survivors of SCLC is emphasized. PMID- 11932359 TI - Temporary stenting for malignant tracheal stenosis due to esophageal cancer: a case report. AB - We present a case of a 56-year-old male with esophageal cancer who underwent successful temporary tracheal stenting followed by a surgical procedure. The trachea was obstructed owing to endotracheal metastasis and the patient had severe dyspnea. A silicone Y-stent was initially inserted into the tracheal stenosis to secure the airway, and then tumor-specific chemo-radiotherapy was applied. The tumor was reduced, the stent was removed and a pathological study indicated that the tracheal metastasis had disappeared. The patient then underwent esophagectomy and tumor-specific chemo-radiotherapy was continued after the surgery. The patient has remained alive and free of esophageal cancer for 18 months after the airway stent emplacement. These findings suggested that the silicone stent was suitable as a temporary measure and that temporary stenting combined with tumor-specific therapy was effective as part of the aggressive therapeutic strategy with which to treat the malignant airway stenosis due to esophageal cancer. PMID- 11932361 TI - Dose-intensive chemotherapy with syngeneic peripheral blood stem cell support for poor risk germ cell tumor of extragonadal origin: a case report. AB - A variety of regimens of high-dose chemotherapy with hematopoietic stem cell support for poor risk germ cell tumors have been established. However, a series of chemotherapy steps carried out prior to the harvest sometimes leads to an insufficient number of peripheral blood stem cells. Here, we report a case of a patient who successfully underwent high-dose chemotherapy for the treatment of poor risk extragonadal germ cell tumor by receiving peripheral blood stem cell transplantation donated from his genomically identical twin brother. PMID- 11932362 TI - De novo appearance of t(7;13)(q10;q33) in the leukemic phase of myelodysplastic syndrome: a case report. AB - Clonal cytogenetic abnormalities are found in about 50% of all patients with myelodysplastic syndrome (MDS) and the clinical implication of these abnormalities is now well documented. However, the de novo appearance of balanced translocations in MDS patients during the progression of the disease is rarely reported and the significance of the balanced translocation remain to be elucidated. We report here the first case of refractory anemia with excess blasts in transformation (RAEBt), in which a new chromosomal translocation, t(7;13)(q10;q33) appeared de novo in the AML phase. It has been revealed that rearrangements and deletions of chromosome 7, i.e. der(1;7)(q10;p10), are very complex and that multiple regions may contribute to the disease phenotype and progression. Our case suggests that the chromosomal region at 7q10, rather than 1p10, might be one of the hot spots for myeloid proliferative disorders, including MDS. PMID- 11932364 TI - Position statement on the use of the ankle-brachial index in the evaluation of patients with peripheral vascular disease: a consensus statement developed by the standards division of the society of cardiovascular & interventional radiology. PMID- 11932363 TI - Citron-kinase, a protein essential to cytokinesis in neuronal progenitors, is deleted in the flathead mutant rat. PMID- 11932365 TI - Peripheral arterial balloon angioplasty: effect of short versus long balloon inflation times on the morphologic results. AB - PURPOSE: To evaluate the effect of different balloon inflation times on angiographic results in peripheral angioplasty. MATERIALS AND METHODS: Seventy four infrainguinal arteriosclerotic lesions were randomized prospectively to undergo balloon dilation for 30 seconds (group I) or 180 seconds (group II). Each group consisted of 37 patients. Postinterventional angiograms were evaluated by two blinded readers. Dissections were graded as follows: 1 = no dissection; 2 = minor flap; 3 = extensive dissection membrane, not flow limiting; or 4 = flow limiting flap. The rate of major-grade dissections (grades 3 and 4), residual stenosis (>30%), and further interventions were compared with the two-tailed chi(2) test. RESULTS: In group I, major dissections were noted in 16 patients (43%) compared with five patients (14%) in group II (P =.009). Residual stenoses were found in 12 patients (32%) in group I compared with five patients (14%) in group II (P =.096). The rate of additional interventions was significantly higher in group I than in group II (20 of 37 vs nine of 37; P =.017). CONCLUSION: A prolonged inflation time of 180 seconds improves the immediate angioplasty result of infrainguinal lesions compared to a short dilation strategy. Significantly fewer major dissections and a modest reduction of residual stenoses are observed. The requirement of costly and time-consuming further interventions is significantly reduced. PMID- 11932366 TI - Failure of prolonged dilation to improve long-term patency of femoropopliteal artery angioplasty: results of a prospective trial. AB - PURPOSE: To determine long-term patency of femoropopliteal artery percutaneous transluminal angioplasty (PTA) in a prospective trial during which prolonged balloon inflation was used for optimization of initial results. MATERIALS AND METHODS: Femoropopliteal PTA was performed in 112 limbs of 97 patients. The mean total length of the treated segments was 7.2 cm (95% CI: 5.99-8.46; median: 5.5 cm). In cases of unsatisfactory primary results after standard dilation for 1-3 minutes, the procedure was continued with prolonged dilation (93 limbs; mean balloon inflation time: 31 min; 95% CI: 24.2-37.7; median: 15 min) with use of the same balloon catheter (77 limbs) or a perfusion balloon catheter (35 limbs). Thirty-four proximal infrapopliteal artery stenoses were treated to improve peripheral runoff and 12 short stents were placed because of flow-limiting dissections. RESULTS: Primary hemodynamic success established by Doppler ultrasound (US) criteria was achieved in 92.9% (104 of 112) of the limbs. Three major complications were encountered; none were related to prolonged balloon inflation. The primary patency rate according to Kaplan-Meier analysis was 42% (+/-5% SE) at 1 year and 39% (+/-5%) at 2 and 3 years. The corresponding secondary patency rates were 51% (+/-5%) and 47% (+/-5%). Large numbers of diseased vessels in the treated limb (four to 10 instead of one to three), eccentric lesions (as opposed to concentric morphology), and additional treated segments (instead of only femoropopliteal lesions) were associated with poorer long-term patency. The duration of balloon dilation was not a determinant of long term patency. CONCLUSION: Although prolonged dilation is safe and feasible in femoropopliteal artery PTA, its routine use is not warranted because it does not result in superior long-term patency rates. PMID- 11932367 TI - Use of a catheter-based system to measure blood flow in hemodialysis grafts during angioplasty procedures. AB - PURPOSE: The goals of this investigation were to evaluate the accuracy and reliability of the Angioflow meter system with use of in vitro and in vivo methods and to compare it to the standard Transonics HD01 system in a clinical setting. MATERIALS AND METHODS: The Angioflow meter system consists of a 6-F endovascular catheter and a laptop computer containing proprietary software for this application. Bench-top testing with use of a flow model was performed to determine the accuracy of the Angioflow meter system. Initial in vivo studies were performed with use of an animal model to assess the endovascular performance of the Angioflow meter system. Subsequently, a human clinical trial was performed to compare the Angioflow meter to the standard Transonics HD01 system. Twenty five patients with dysfunctional (<600 mL/min) hemodialysis grafts were referred for fistulography and angioplasty. Intragraft blood flow measurements were obtained before and after angioplasty with use of both the Angioflow meter system and the Transonics HD01 system. A comparison of the two systems was performed. RESULTS: Bench-top testing and animal studies demonstrated an excellent (r =.98) correlation between the measurements of the Angioflow meter and volumetric flow measurements. In the clinical trial, there was reasonable correlation (r =.72) between the blood flow measurements obtained with use of the Angioflow meter and Transonics HD01 system. The reproducibility of consecutive measurements with the Angioflow meter was excellent (r =.98). The mean increase in intragraft blood flow after angioplasty was 320 mL/min. CONCLUSION: The Angioflow meter is an accurate and reliable endovascular device for measuring intragraft blood flow during interventional procedures. Use of this catheter-based system should prove beneficial for quantifying the success of endovascular interventions, the assessment of arterial inflow, and identification of inconspicuous lesions. PMID- 11932368 TI - Placement of a permanent tunneled peritoneal drainage catheter for palliation of malignant ascites: a simplified percutaneous approach. AB - PURPOSE: To report a simple, minimally invasive method of palliative drainage of symptomatic malignant ascites, which allows patients to avoid repeated trips to the hospital for paracentesis. MATERIALS AND METHODS: Since 1996, 29 patients with symptomatic, large-volume malignant ascites underwent percutaneous placement of a tunneled multiple-side-hole Tenckhoff catheter with use of a modified Seldinger technique employing curved and straight coaxial needles. Unlike previously described methods of tunneled catheter placement, this method does not require an incision. RESULTS: All patients were able to drain their ascites at home and all achieved significant improvement in their symptoms attributable to ascites. Twenty-three of the 29 were outpatients and were able to leave the hospital within several hours of catheter placement. Inpatients remained in the hospital for various reasons unrelated to catheter placement and most left within several days. Only one patient died in the hospital during the same admission. One patient developed cellulitis, one had persistent leakage around the catheter, one catheter had to be replaced, and one catheter was accidentally removed at home. No patient developed clinical symptoms of peritonitis or sepsis. All deaths were attributable to patients' underlying malignancies. CONCLUSION: Percutaneous placement of a permanent tunneled catheter using a modified Seldinger technique employing curved and straight coaxial needles is a safe, simple, and effective method for palliative drainage of malignant ascites that allows patients to return home quickly. PMID- 11932369 TI - Safety and efficacy of transjugular intrahepatic portosystemic shunt creation for the treatment of hepatic hydrothorax. AB - PURPOSE: To evaluate safety and efficacy of transjugular intrahepatic portosystemic shunt (TIPS) creation for hepatic hydrothorax (HHyd). MATERIALS AND METHODS: Twenty-one patients underwent TIPS creation for HHyd. A prospective TIPS database and medical records were reviewed. Clinical and radiographic outcomes were recorded as complete (symptom/effusion resolution), partial (improved symptoms/effusion), or none. Data patterns were examined with chi(2) tests and Kaplan-Meier analysis. RESULTS: Patients included 12 women and nine men, with a mean age of 56 years, all with Child class B (n = 7) or C (n = 14) disease. The technical success rate was 100%. Mean follow-up was 223 days. Twenty-nine percent (six of 21) died within 30 days of TIPS creation, 10% (two of 21) underwent transplantation within 30 days, and 62% (13 of 21) survived beyond 30 days. Data were incomplete in two patients. Clinical response was classified as complete in 63% (12 of 19), partial in 11% (two of 19), and none in 26% (five of 19). Radiographic response was classified as complete in 30% (six of 20), partial in 50% (10 of 20), and none in 20% (four of 20). Nonresponders had multisystem organ failure, and all but one died within 30 days. However, of the 13 patients surviving longer than 30 days, 10 (77%) had a complete clinical response. CONCLUSION: TIPS is a relatively safe and effective method of controlling HHyd. The majority of patients experienced improvement or resolution of clinical symptoms with a variable reduction in the quantity of pleural fluid. There was a tendency among nonresponders to die within 30 days. PMID- 11932370 TI - Comparison of four techniques to estimate radiation dose to skin during angiographic and interventional radiology procedures. AB - PURPOSE: Four techniques used to estimate radiation risk were compared to determine whether commonly used dosimetry measurements permit reliable estimates of skin dose. Peak skin dose (PSD) is known to be the most reliable estimate of risk to skin. The purpose of this study is to determine peak skin dose with use of real-time software measurements and to correlate other measures of dose with PSD. MATERIALS AND METHODS: Two hundred twelve patients undergoing arch aortography and bilateral carotid arteriography (referred to as "carotid"), abdominal aortography and bilateral lower extremity runoff ("runoff"), or tunneled chest wall port placement ("port") were studied. Fluoroscopy time, dose area product (DAP), and cumulative dose at the interventional reference point were recorded for all procedures; PSD was recorded for a subset of 105 procedures. The dose index, defined as the ratio between PSD and cumulative dose, was also determined. RESULTS: In general, correlation values for comparisons between fluoroscopy time and the other measures of dose (r =.29 to.78) were lower than values for comparisons among DAP, cumulative dose, and PSD (r =.52 to.94). For all procedures, pair-wise correlations between DAP, cumulative skin dose, and PSD were statistically significant (P <.01) The ratio between PSD and cumulative skin dose (dose index) was significantly different for ports versus other procedures (carotid, Z = 4.62, P <.001; runoff, Z = 4.52, P <.001), but carotid and runoff procedures did not differ significantly in this regard (Z = 0.746, P =.22). Within each individual procedure type, the range of values for the dose index varied 156.7-fold for carotid arteriography, 3.2-fold for chest ports, and 175-fold for aortography and runoff. CONCLUSION: Fluoroscopy time is a poor predictor of risk because it does not correlate well with PSD. Cumulative dose and DAP are not good analogues of PSD because of weak correlations for some procedures and because of wide variations in the dose index for all procedures. PMID- 11932371 TI - Occlusion of the fallopian tube by selective transcatheter radiofrequency electrocoagulation: an experimental study in rabbits. AB - PURPOSE: To test the feasibility of performing transcervical fallopian tube occlusion in a rabbit model with use of unipolar radiofrequency (RF) electrocoagulation. MATERIALS AND METHODS: Under fluoroscopic guidance, transvaginal catheterization of the right or left fallopian tube was first performed with use of a coaxial technique in 20 rabbits. With a metal guide wire protruding from the catheter serving as the active electrode, RF electrocoagulation was performed. The power output was set to 200 W and the current was applied for 20 seconds. The fallopian tube on the contralateral side and the uterus were used as controls. Rabbits were randomly designated to be killed either 2 days (group I, n = 10) or 30 days (group II, n = 10) after the procedure, and tubal patency and histologic changes were evaluated. RESULTS: In group I, significant necrosis of the endosalpinx caused by RF electrocoagulation was found in all 10 rabbits. In group II, the occluded fallopian tube showed fibrosis of the wall in all 10 rabbits, but there was no tissue damage to adjacent organs. Histologic findings in the contralateral fallopian tubes were normal in all 20 rabbits. CONCLUSION: Transcatheter RF electrocoagulation proved to be a safe nonsurgical alternative for occlusion of the fallopian tubes in rabbits. PMID- 11932372 TI - Use of balloon-expandable stents in transjugular intrahepatic portosystemic shunts in cases of Wallstent endoprosthesis technical failure and revision of shunt stenosis. AB - Thirteen patients underwent placement of a balloon-expandable stent either at initial transjugular intrahepatic portosystemic shunt (TIPS) creation (n = 3) because of immediate technical failure of the Wallstent or at shunt revision because of failure of the Wallstent to reduce the portosystemic gradient 2-fold at 16 h after rotavirus infection, and only one gene was similarly regulated at 1 h postinfection. Of these transcriptional changes, 73% corresponded to the upregulation of genes, with the majority of them occurring late, at 12 or more hours postinfection. Some of the regulated genes were classified according to known biological function and included genes encoding integral membrane proteins, interferon-regulated genes, transcriptional and translational regulators, and calcium metabolism-related genes. A new picture of global transcriptional regulation in the infected cell is presented and families of genes which may be involved in viral pathogenesis are discussed. PMID- 11932414 TI - Reovirus core protein mu2 determines the filamentous morphology of viral inclusion bodies by interacting with and stabilizing microtubules. AB - Cells infected with mammalian reoviruses often contain large perinuclear inclusion bodies, or "factories," where viral replication and assembly are thought to occur. Here, we report a viral strain difference in the morphology of these inclusions: filamentous inclusions formed in cells infected with reovirus type 1 Lang (T1L), whereas globular inclusions formed in cells infected with our laboratory's isolate of reovirus type 3 Dearing (T3D). Examination by immunofluorescence microscopy revealed the filamentous inclusions to be colinear with microtubules (MTs). The filamentous distribution was dependent on an intact MT network, as depolymerization of MTs early after infection caused globular inclusions to form. The inclusion phenotypes of T1L x T3D reassortant viruses identified the viral M1 genome segment as the primary genetic determinant of the strain difference in inclusion morphology. Filamentous inclusions were seen with 21 of 22 other reovirus strains, including an isolate of T3D obtained from another laboratory. When the mu2 proteins derived from T1L and the other laboratory's T3D isolate were expressed after transfection of their cloned M1 genes, they associated with filamentous structures that colocalized with MTs, whereas the mu2 protein derived from our laboratory's T3D isolate did not. MTs were stabilized in cells infected with the viruses that induced filamentous inclusions and after transfection with the M1 genes derived from those viruses. Evidence for MT stabilization included bundling and hyperacetylation of alpha tubulin, changes characteristically seen when MT-associated proteins (MAPs) are overexpressed. Sequencing of the M1 segments from the different T1L and T3D isolates revealed that a single-amino-acid difference at position 208 correlated with the inclusion morphology. Two mutant forms of mu2 with the changes Pro-208 to Ser in a background of T1L mu2 and Ser-208 to Pro in a background of T3D mu2 had MT association phenotypes opposite to those of the respective wild-type proteins. We conclude that the mu2 protein of most reovirus strains is a viral MAP and that it plays a key role in the formation and structural organization of reovirus inclusion bodies. PMID- 11932415 TI - Immune-mediated protection from measles virus-induced central nervous system disease is noncytolytic and gamma interferon dependent. AB - Neurons of the mammalian central nervous system (CNS) are an essential and largely nonrenewable cell population. Thus, virus infections that result in neuronal depletion, either by virus-mediated cell death or by induction of the cytolytic immune response, could cause permanent neurological impairment of the host. In a transgenic mouse model of measles virus (MV) infection of neurons, we have previously shown that the host T-cell response was required for resolution of infection in susceptible adult mice. In this report, we show that this protective response did not result in neuronal death, even during the peak of T cell infiltration into the brain parenchyma. When susceptible mice were intercrossed with specific immune knockout mice, a critical role for gamma interferon (IFN-gamma) was identified in protection against MV infection and CNS disease. Moreover, the addition of previously activated splenocytes or recombinant murine IFN-gamma to MV-infected primary neurons resulted in the inhibition of viral replication in the absence of neuronal death. Together, these data support the hypothesis that the host immune response can promote viral clearance without concomitant neuronal loss, a process that appears to be mediated by cytokines. PMID- 11932417 TI - Promotion of alpha/beta interferon induction during in vivo viral infection through alpha/beta interferon receptor/STAT1 system-dependent and -independent pathways. AB - Viruses and viral components can be potent inducers of alpha/beta interferons (IFN-alpha/beta). In culture, IFN-alpha/beta prime for their own expression, in response to viruses, through interferon regulatory factor 7 (IRF-7) induction. The studies presented here evaluated the requirements for functional IFN receptors and the IFN signaling molecule STAT1 in IFN-alpha/beta induction during infections of mice with lymphocytic choriomeningitis virus (LCMV). At 24 h after infection, levels of induced IFN-alpha/beta in serum were reduced 90 to 95% in IFN-alpha/beta receptor-deficient (IFN-alpha/betaR(-/-)) and STAT1(-/-) mice compared to those in wild-type mice. However, at 48 h, these mice showed elevated expression in the serum whereas IFN-alpha/beta levels were still reduced >75% in IFN-alpha/betagammaR(-/-) mice even though the viral burden was heavy. Levels of IFN-beta, IFN-alpha4, and non-IFN-alpha4 subtype mRNA expression correlated with IFN-alpha/beta bioactivity, and all IFN-alpha/beta subtypes were coincidentally detectable. IRF-7 mRNA was induced under conditions of IFN-alpha/beta production, including late production in IFN-alpha/betaR(-/-) mice. These data demonstrate that the presence of the virus alone is not sufficient to induce IFN-alpha/beta during LCMV infection in vivo. Instead, autocrine amplification through the IFN alpha/betaR is necessary for optimal induction. In the absence of a functional IFN-alpha/betaR, however, alternative mechanisms, independent of STAT1 but requiring a functional IFN-gammaR, take over. PMID- 11932416 TI - Effects of mutations in the adenoviral E1B 55-kilodalton protein coding sequence on viral late mRNA metabolism. AB - The human subgroup C adenoviral E1B 55-kDa protein cooperates with the viral E4 Orf6 protein to induce selective export of viral, late mRNAs from the nucleus to the cytoplasm. Previous studies have suggested that such preferential transport of viral mRNA and the concomitant inhibition of export of cellular mRNAs are the result of viral colonization of specialized microenvironments within the nucleus. However, neither the molecular basis of this phenomenon nor the mechanism by which the E1B 55-kDa protein acts has been elucidated. We therefore examined viral late mRNA metabolism in HeLa cells infected with a series of mutant viruses that carry insertions at various positions in the E1B protein coding sequence (P. R. Yew, C. C. Kao, and A. J. Berk, Virology 179:795-805, 1990). All the mutations examined impaired cytoplasmic accumulation of viral L2 mRNAs and reduced L2 mRNA export efficiency. However, in most cases these defects could be ascribed to reduced E1B 55-kDa protein concentration or the unexpected failure of the altered E1B proteins to enter the nucleus efficiently. The latter property, the pleiotropic defects associated with all the mutations that impaired nuclear entry of the E1B protein, and consideration of its primary sequence suggest that these insertions result in misfolding of the protein. Insertion of four amino acids at residue 143 also inhibited viral mRNA export but resulted in increased rather than decreased accumulation of the E1B 55-kDa protein in the nucleus. This mutation specifically impaired the previously described association of the E1B protein with intranuclear structures that correspond to sites of adenoviral DNA replication and transcription (D. Ornelles and T. Shenk, J. Virol. 65:424-439, 1991) and the colocalization of the E1B and E4 Orf6 proteins. As this insertion has been shown to inhibit the interaction of the E1B with the E4 Orf6 protein in infected cell extracts (S. Rubenwolf, H. Schutt, M. Nevels, H. Wolf, and T. Dobner, J. Virol. 71:1115-1123, 1997), these phenotypes provide direct support for the hypothesis that selective viral mRNA export is determined by the functional organization of the infected cell nucleus. PMID- 11932418 TI - Expression of exogenous Sam68, the 68-kilodalton SRC-associated protein in mitosis, is able to alleviate impaired Rev function in astrocytes. AB - Human immunodeficiency virus type 1 (HIV-1) gene expression in astrocytes is restricted, resulting in a brief and limited synthesis of HIV-1 viral structural proteins. Impaired Rev function has been documented in these cells. However, the molecular mechanisms underlying the impaired Rev function are not fully understood. Using the astroglial cell line U87.MG as a model, we report here that HIV-1 gene expression down-regulated expression of Sam68, the 68-kDa Src associated protein in mitosis, which was constitutively expressed at a lower level in astrocytes. Elevating the endogenous level of Sam68 expression considerably restored HIV-1 Rev function in astrocytes, as determined by a Rev dependent reporter gene assay. However, elevation of Sam68 expression achieved only a modest increase in HIV-1 production, further supporting the notion that there are multiple cellular restrictions of HIV-1 gene expression in astrocytes. Mutagenesis analysis identified the region between amino acids 321 and 410 of Sam68 as being directly involved in the binding of Sam68 to Rev, while a double mutation in Rev, L78D and E79L, like those in the dominant-negative Rev mutant M10, eliminated Rev binding to Sam68. Moreover, subcellular fractionation and digital fluorescence microscopic imaging revealed that Sam68 expression promoted Rev nuclear export. Taken together, our studies demonstrate that a lower level of constitutive Sam68 expression, followed by further down-regulation by HIV-1 infection, contributes to impaired Rev function in astrocytes, and they suggest that Sam68 may play an important role in Rev nuclear export. PMID- 11932419 TI - Plasmid vectors encoding cholera toxin or the heat-labile enterotoxin from Escherichia coli are strong adjuvants for DNA vaccines. AB - Two plasmid vectors encoding the A and B subunits of cholera toxin (CT) and two additional vectors encoding the A and B subunits of the Escherichia coli heat labile enterotoxin (LT) were evaluated for their ability to serve as genetic adjuvants for particle-mediated DNA vaccines administered to the epidermis of laboratory animals. Both the CT and the LT vectors strongly augmented Th1 cytokine responses (gamma interferon [IFN-gamma]) to multiple viral antigens when codelivered with DNA vaccines. In addition, Th2 cytokine responses (interleukin 4 [IL-4]) were also augmented by both sets of vectors, with the effects of the LT vectors on IL-4 responses being more antigen dependent. The activities of both sets of vectors on antibody responses were antigen dependent and ranged from no effect to sharp reductions in the immunoglobulin G1 (IgG1)-to-IgG2a ratios. Overall, the LT vectors exhibited stronger adjuvant effects in terms of T-cell responses than did the CT vectors, and this was correlated with the induction of greater levels of cyclic AMP by the LT vectors following vector transfection into cultured cells. The adjuvant effects observed in vivo were due to the biological effects of the encoded proteins and not due to CpG motifs in the bacterial genes. Interestingly, the individual LT A and B subunit vectors exhibited partial adjuvant activity that was strongly influenced by the presence or absence of signal peptide coding sequences directing the encoded subunit to either intracellular or extracellular locations. Particle-mediated delivery of either the CT or LT adjuvant vectors in rodents and domestic pigs was well tolerated, suggesting that bacterial toxin-based genetic adjuvants may be a safe and effective strategy to enhance the potency of both prophylactic and therapeutic DNA vaccines for the induction of strong cellular immunity. PMID- 11932420 TI - Bak and Bax function to limit adenovirus replication through apoptosis induction. AB - Adenovirus infection and expression of E1A induces both proliferation and apoptosis, the latter of which is blocked by the adenovirus Bcl-2 homologue E1B 19K. The mechanism of apoptosis induction and the role that it plays in productive infection are not known. Unlike apoptosis mediated by death receptors, infection with proapoptotic E1B 19K mutant viruses did not induce cleavage of Bid but nonetheless induced changes in Bak and Bax conformation, Bak-Bax interaction, caspase 9 and 3 activation, and apoptosis. In wild-type-adenovirus-infected cells, in which E1B 19K inhibits apoptosis, E1B 19K was bound to Bak, precluding Bak-Bax interaction and changes in Bax conformation. Infection with E1B 19K mutant viruses induced apoptosis in wild-type and Bax- or Bak-deficient baby mouse kidney cells but not in those deficient for both Bax and Bak. Furthermore, Bax and Bak deficiency dramatically increased E1A expression and virus replication. Thus, Bax- and Bak-mediated apoptosis severely limits adenoviral replication, demonstrating that Bax and Bak function as an antiviral response at the cellular level. PMID- 11932421 TI - Adeno-associated virus capsids displaying immunoglobulin-binding domains permit antibody-mediated vector retargeting to specific cell surface receptors. AB - Recombinant adeno-associated virus type 2 (rAAV2) is a promising vector for human somatic gene therapy. However, its broad host range is a disadvantage for some applications, because it reduces the specificity of the gene transfer. To overcome this limitation, we sought to create a versatile rAAV vector targeting system which would allow us to redirect rAAV binding to specific cell surface receptors by simple coupling of different ligands to its capsid. For this purpose, an immunoglobulin G (IgG) binding domain of protein A, Z34C, was inserted into the AAV2 capsid at amino acid position 587. The resulting AAV2-Z34C mutants could be packaged and purified to high titers and bound to IgG molecules. rAAV2-Z34C vectors coupled to antibodies against CD29 (beta(1)-integrin), CD117 (c-kit receptor), and CXCR4 specifically transduced distinct human hematopoietic cell lines. In marked contrast, no transduction was seen in the absence of antibodies or in the presence of specific blocking reagents. These results demonstrate for the first time that an immunoglobulin binding domain can be inserted into the AAV2 capsid and coupled to various antibodies, which mediate the retargeting of rAAV vectors to specific cell surface receptors. PMID- 11932423 TI - Differential activation of innate immune responses by adenovirus and adeno associated virus vectors. AB - Adenovirus vectors induce acute inflammation of infected tissues due to activation of the innate immune system and expression of numerous chemokines and cytokines in transduced target cells. In contrast, adeno-associated virus (AAV) vectors are not associated with significant inflammation experimentally or clinically. We tested the ability of AAV vectors to induce the expression of chemokines in vitro and to activate the innate immune system in vivo. In human HeLa cells and murine renal epithelium-derived cells (REC cells) the adenovirus vector AdlacZ induced the expression of multiple inflammatory chemokines including RANTES, interferon-inducible protein 10 (IP-10), interleukin-8 (IL-8), MIP-1beta, and MIP-2 in a dose-dependent manner. The use of AAVlacZ did not induce the expression of these chemokines above baseline levels despite 40-fold greater titers than AdlacZ and greater amounts of intracellular AAVlacZ genomes according to Southern and slot blot analysis. This finding confirmed that the lack of AAVlacZ induction of chemokine was not due to reduced transduction. In DBA/2 mice, the intravenous administration of 2.5 x 10(11) particles of AAVlacZ resulted in the rapid induction of liver tumor necrosis factor alpha (TNF-alpha), RANTES, IP-10, MIP-1beta, MCP-1, and MIP-2 mRNAs. However, 6 h following injection, chemokine mRNA levels returned to baseline. As expected, administration of 10-fold less AdlacZ caused an induction of liver TNF-alpha and chemokine mRNAs that persisted for more than 24 h posttransduction. Whereas intravenous administration of 2.5 x 10(11) particles of AAVlacZ triggered a transient infiltration of neutrophils and CD11b(+) cells into liver, this response stood in contrast to widespread inflammation and toxicity induced by AdlacZ. Kupffer cell depletion abolished AAVlacZ but not AdlacZ-induced chemokine expression and neutrophil infiltration. In summary, these results show that AAV vectors activate the innate immune system to a lesser extent than do adenovirus vectors and offer a possible explanation for the reduced inflammatory properties of AAV compared to adenovirus vectors. PMID- 11932422 TI - The oncogenic protein kinase Tpl-2/Cot contributes to Epstein-Barr virus-encoded latent infection membrane protein 1-induced NF-kappaB signaling downstream of TRAF2. AB - The Epstein-Barr virus-encoded latent infection membrane protein 1 (LMP1) is a pleiotropic protein, the activities of which include effects on cell transformation and phenotype, growth, and survival. The ability of LMP1 to mediate at least some of these phenomena could be attributed to the activation of the transcription factor NF-kappaB. LMP1 promotes NF-kappaB activation through the recruitment of the adapter protein TRAF2 and the formation of a dynamic multiprotein complex that includes the NF-kappaB kinase, the IkappaB kinases, and their downstream targets, IkappaBs and p105. In this study, we have identified the oncogenic kinase Tpl-2/Cot as a novel component of LMP1-induced NF-kappaB signaling. We show that Tpl-2 is expressed in primary biopsies from patients with nasopharyngeal carcinoma and Hodgkin's disease, where LMP1 is also found. Inducible expression of LMP1 promotes the activation of Tpl-2, and a catalytically inactive Tpl-2 mutant suppresses LMP1-induced NF-kappaB signaling. In colocalization and coimmunoprecipitation experiments, Tpl-2 and TRAF2 were found to interact with Tpl-2 functioning downstream of TRAF2. Consistent with this observation, catalytically inactive Tpl-2 also blocked CD40-mediated NF kappaB activation, which largely depends on TRAF2. The ability of Tpl-2 to influence LMP1-induced NF-kappaB occurs through modulation of both IkappaBalpha and p105 functions. Furthermore, Tpl-2 was found to influence the expression of angiogenic mediators, such as COX-2 in LMP1-transfected cells. These data identify Tpl-2 as a component of LMP1 signaling downstream of TRAF2 and as a modulator of LMP1-mediated effects. PMID- 11932425 TI - Fatty acids on the A/USSR/77 influenza virus hemagglutinin facilitate the transition from hemifusion to fusion pore formation. AB - Influenza virus hemagglutinin (HA) has three highly conserved acylation sites close to the carboxyl terminus of the HA2 subunit, one in the transmembrane domain and two in the cytoplasmic domain. Each site is modified by palmitic acid through a thioester linkage to cysteine. To elucidate the biological significance of HA acylation, the acylation sites of HA of influenza virus strain A/USSR/77 (H1N1) were changed by site-directed mutagenesis, and the membrane fusion activity of mutant HAs lacking the acylation site(s) was examined quantitatively using transfer assays of lipid (R18) and aqueous (calcein) dyes. Lipid mixing, so called hemifusion, activity was not affected by deacylation, whereas transfer of aqueous dye, so-called fusion pore formation, was dramatically restricted. When the fusion reaction was induced by a lower pH than the optimal one, calcein transfer with the mutant HAs was improved, but simultaneously a considerable calcein leakage into the medium was observed. From these results, we conclude that the palmitic acids on the H1 subtype HA facilitate the transition from hemifusion to fusion pore formation. PMID- 11932424 TI - Rafts promote assembly and atypical targeting of a nonenveloped virus, rotavirus, in Caco-2 cells. AB - Rotavirus follows an atypical pathway to the apical membrane of intestinal cells that bypasses the Golgi. The involvement of rafts in this process was explored here. VP4 is the most peripheral protein of the triple-layered structure of this nonenveloped virus. High proportions of VP4 associated with rafts within the cell as early as 3 h postinfection. In the meantime a significant part of VP4 was targeted to the Triton X-100-resistant microdomains of the apical membrane, suggesting that this protein possesses an autonomous signal for its targeting. At a later stage the other structural rotavirus proteins were also found in rafts within the cells together with NSP4, a nonstructural protein required for the final stage of virus assembly. Rafts purified from infected cells were shown to contain infectious particles. Finally purified VP4 and mature virus were shown to interact with cholesterol- and sphingolipid-enriched model lipid membranes that changed their phase preference from inverted hexagonal to lamellar structures. Together these results indicate that a direct interaction of VP4 with rafts promotes assembly and atypical targeting of rotavirus in intestinal cells. PMID- 11932427 TI - The carboxy terminus of simian virus 40 large T antigen is required to disrupt the yeast cell cycle. AB - Wild-type and J domain mutant simian virus 40 large T antigens alter the cell cycle and bud morphology of Saccharomyces cerevisiae. In contrast, yeast cells expressing mutant T antigen lacking the carboxy-terminal 150 aa exhibit normal morphology, indicating that this region of T antigen is required for cell cycle disruption. PMID- 11932426 TI - Exploiting the natural diversity in adenovirus tropism for therapy and prevention of disease. AB - Since targeting of recombinant adenovirus vectors to defined cell types in vivo is a major challenge in gene therapy and vaccinology, we explored the natural diversity in human adenovirus tissue tropism. Hereto, we constructed a library of Ad5 vectors carrying fibers from other human serotypes. From this library, we identified vectors that efficiently infect human cells that are important for diverse gene therapy approaches and for induction of immunity. For several medical applications (prenatal diagnosis, artificial bone, vaccination, and cardiovascular disease), we demonstrate the applicability of these novel vectors. In addition, screening cell types derived from different species revealed that cellular receptors for human subgroup B adenoviruses are not conserved between rodents and primates. These results provide a rationale for utilizing elements of human adenovirus serotypes to generate chimeric vectors that improve our knowledge concerning adenovirus biology and widen the therapeutic window for vaccination and many different gene transfer applications. PMID- 11932428 TI - Nef-mediated downregulation of CD4 enhances human immunodeficiency virus type 1 replication in primary T lymphocytes. AB - The accessory protein Nef plays a crucial role in primate lentivirus pathogenesis. Nef enhances human immunodeficiency virus type 1 (HIV-1) infectivity in culture and stimulates viral replication in primary T cells. In this study, we investigated the relationship between HIV-1 replication efficiency in CD4(+) T cells purified from human blood and two various known activities of Nef, CD4 downregulation and single-cycle infectivity enhancement. Using a battery of reporter viruses containing point mutations in nef, we observed a strong genetic correlation between CD4 downregulation by Nef during acute HIV-1 infection of activated T cells and HIV-1 replication efficiency in T cells. In contrast, HIV-1 replication ability was not significantly correlated with the ability of Nef to enhance single-cycle virion infectivity, as determined by using viruses produced in cells lacking CD4. These results demonstrate that CD4 downregulation by Nef plays a crucial role in HIV-1 replication in activated T cells and underscore the potential for the development of therapies targeting this conserved activity of Nef. PMID- 11932429 TI - Highly stable trimers formed by human immunodeficiency virus type 1 envelope glycoproteins fused with the trimeric motif of T4 bacteriophage fibritin. AB - The envelope glycoproteins of human immunodeficiency virus type 1 (HIV-1) function as a trimer composed of three gp120 exterior glycoproteins and three gp41 transmembrane proteins. Soluble gp140 glycoproteins composed of the uncleaved ectodomains of gp120 and gp41 form unstable, heterogeneous oligomers, but soluble gp140 trimers can be stabilized by fusion with a C-terminal, trimeric GCN4 motif (X. Yang et al., J. Virol. 74:5716-5725, 2000). To understand the influence of the C-terminal trimerization domain on the properties of soluble HIV 1 envelope glycoprotein trimers, uncleaved, soluble gp140 glycoproteins were stabilized by fusion with another trimeric motif derived from T4 bacteriophage fibritin. The fibritin construct was more stable to heat and reducing conditions than the GCN4 construct. Both GCN4- and fibritin-stabilized soluble gp140 glycoproteins exhibited patterns of neutralizing and nonneutralizing antibody binding expected for the functional envelope glycoprotein spike. Of note, two potently neutralizing antibodies, immunoglobulin G1b12 and 2G12, exhibited the greatest recognition of the stabilized, soluble trimers, relative to recognition of the gp120 monomer. The observed similarities between the GCN4 and fibritin constructs indicate that the HIV-1 envelope glycoprotein ectodomains dictate many of the antigenic and structural features of these fusion proteins. The melting temperatures and ligand recognition properties of the GCN4- and fibritin stabilized soluble gp140 glycoproteins suggest that these molecules assume conformations distinct from that of the fusion-active, six-helix bundle. PMID- 11932431 TI - Scrapie infectivity in hamster blood is not associated with platelets. AB - The infectivity of hamster scrapie strain 263K was measured in platelets isolated from blood pooled from six hamsters with clinical scrapie. The total number of infectious doses present in the blood pool was 220, out of which only 3.5 infectious doses were associated with platelets. A larger proportion of the total infectivity was recovered from the mononuclear leukocyte fraction. This result indicates that platelets are not the source of blood-borne infectivity in transmissible spongiform encephalopathy-infected hamsters. PMID- 11932430 TI - Functional analysis of the murine sarcoma virus RNA packaging sequence. AB - We investigated the features of the Moloney murine sarcoma virus leader sequence necessary for RNA packaging function by using a deletion analysis approach. We found that sequences that extend beyond those characterized genetically in previous reports are important for optimal packaging efficiency. A fragment covering a minimum of four potential stem-loop structures is required for the shortest packaging element compatible with gene transfer. Our results reveal the extent to which each of the segments of the packaging sequence contribute to packaging efficiency. PMID- 11932432 TI - Characterization and complete nucleotide sequence of an unusual reptilian retrovirus recovered from the order Crocodylia. AB - A novel group of retroviruses found within the order Crocodylia are described. Phylogenetic analyses demonstrate that they are probably the most divergent members of the Retroviridae described to date; even the most conserved regions of Pol show an average of only 23% amino acid identity when compared to other retroviruses. PMID- 11932433 TI - Stabilization of a full-length infectious cDNA clone of transmissible gastroenteritis coronavirus by insertion of an intron. AB - The stable propagation of a full-length transmissible gastroenteritis coronavirus (TGEV) cDNA in Escherichia coli cells as a bacterial artificial chromosome has been considerably improved by the insertion of an intron to disrupt a toxic region identified in the viral genome. The viral RNA was expressed in the cell nucleus under the control of the cytomegalovirus promoter and the intron was efficiently removed during translocation of this RNA to the cytoplasm. The insertion in two different positions allowed stable plasmid amplification for at least 200 generations. Infectious TGEV was efficiently recovered from cells transfected with the modified cDNAs. PMID- 11932434 TI - Dengue type 3 virus in plasma is a population of closely related genomes: quasispecies. AB - Using reverse transcription-PCR and clonal sequencing of the dengue virus envelope gene derived from the plasma samples of six patients, we reported for the first time that dengue virus circulates as a population of closely related genomes. The extent of sequence diversity varied among patients, with the mean pairwise proportions of difference ranging from 0.21 to 1.67%. Genome-defective viruses were found in 5.8% of the total number of clones analyzed. Our findings on the quasispecies nature of dengue virus and the defective virus in vivo have implications with regard to the pathogenesis of dengue virus. PMID- 11932435 TI - Specific incorporation of heat shock protein 70 family members into primate lentiviral virions. AB - To determine if any heat shock proteins are incorporated into human immunodeficiency virus type 1 (HIV-1) virions in a manner similar to that of the peptidyl-prolyl isomerase cyclophilin A, we probed purified virions with antibodies against heat shock proteins Hsp27, Hsp40, Hsp60, Hsp70, Hsc70, and Hsp90. Of these proteins, Hsp60, Hsp70, and Hsc70 associated with virions purified based on either particle density or size and were shown to be incorporated within the virion membrane, where they were protected from digestion by exogenous protease. Virion incorporation of Hsp70 was also observed with HIV-2 and with simian immunodeficiency viruses SIV(MAC) and SIV(AGM), but it appears to be specific for primate lentiviruses, since Hsp70 was not detected in association with Moloney murine leukemia virus virions. Of the HIV-1 genes, gag was found to be sufficient for Hsp70 incorporation, though Hsp70 was roughly equimolar with pol-encoded proteins in virions. PMID- 11932436 TI - Cyclophilin a plays distinct roles in human immunodeficiency virus type 1 entry and postentry events, as revealed by spinoculation. AB - Cyclophilin A (CypA) is necessary for effective human immunodeficiency virus type 1 (HIV-1) replication. However, the functions of CypA and the precise steps at which CypA acts in the HIV-1 life cycle remain to be determined. By using a methodology that bypasses the need for attachment factors-spinoculation-we present evidence that CypA participates in both entry and postentry events. PMID- 11932437 TI - Sensing, signalling and integrating physical processes during Saccharomyces cerevisiae invasive and filamentous growth. PMID- 11932438 TI - LuxS: its role in central metabolism and the in vitro synthesis of 4-hydroxy-5 methyl-3(2H)-furanone. AB - Many bacteria produce extracellular molecules which function in cell-to-cell communication. One of these molecules, autoinducer 2 (AI-2), was first described as an extracellular signal produced by Vibrio harveyi to control luciferase expression. Subsequently, a number of bacteria have been shown to possess AI-2 activity in their culture supernatants, and bear the luxS gene product, which is required for AI-2 synthesis. In Porphyromonas gingivalis, luxS and pfs, encoding a 5'-methylthioadenosine/S-adenosylhomocysteine nucleosidase (MTA/SAH'ase), form an operon, suggesting that S-adenosylhomocysteine (SAH) or 5'-methylthioadenosine (MTA) serves as a substrate for AI-2 production. Cell-free extracts of Escherichia coli MG1655, but not DH5alpha (which carries a luxS frame-shift mutation) were capable of generating AI-2 activity upon addition of SAH, but not MTA. S-Ribosyl-homocysteine (RH) derived from SAH also served as a substrate in E. coli MG1655 extracts. RH-supplemented cell-free extracts of Pseudomonas aeruginosa, a bacterium that lacks luxS, only generated AI-2 activity following the introduction of a plasmid containing the Por. gingivalis pfs-luxS operon. In addition, defined in vitro systems consisting of the purified LuxS proteins from Por. gingivalis, E. coli, Neisseria meningitidis or Staphylococcus aureus converted RH to homocysteine and a compound that exhibits AI-2 activity.4-Hydroxy 5-methyl-3(2H)-furanone was identified by mass spectrometry analysis as a major product formed in this in vitro reaction. In E. coli MG1655, expression of T3SH [the bacteriophage T3 S-adenosylmethionine (SAM) hydrolase] significantly reduced AI-2 activity in culture supernatants, suggesting that AI-2 production is limited by the amount of SAH produced in SAM-dependent transmethylase reactions. The authors suggest that the LuxS protein has an important metabolic function in the recycling of SAH. They also show that Ps. aeruginosa is capable of removing AI-2 activity, implying that this molecule may act as a nutrient. In many bacteria AI 2 may in fact represent not a signal molecule but a metabolite which is released early and metabolized in the later stages of growth. PMID- 11932439 TI - Genetically programmed autoinducer destruction reduces virulence gene expression and swarming motility in Pseudomonas aeruginosa PAO1. AB - Virulence in the opportunistic human pathogen Pseudomonas aeruginosa is controlled by cell density via diffusible signalling molecules ('autoinducers') of the N-acylhomoserine lactone (AHL) type. Two Bacillus sp. isolates (A23 and A24) with AHL-degrading activity were identified among a large collection of rhizosphere bacteria. From isolate A24 a gene was cloned which was similar to the aiiA gene, encoding an AHL lactonase in another Bacillus strain. Expression of the aiiA homologue from isolate A24 in P. aeruginosa PAO1 reduced the amount of the quorum sensing signal N-oxododecanoyl-L-homoserine lactone and completely prevented the accumulation of the second AHL signal, N-butyryl-L-homoserine lactone. This strongly reduced AHL content correlated with a markedly decreased expression and production of several virulence factors and cytotoxic compounds such as elastase, rhamnolipids, hydrogen cyanide and pyocyanin, and strongly reduced swarming. However, no effect was observed on flagellar swimming or on twitching motility, and aiiA expression did not affect bacterial adhesion to a polyvinylchloride surface. In conclusion, introduction of an AHL degradation gene into P. aeruginosa could block cell-cell communication and exoproduct formation, but failed to interfere with surface colonization. PMID- 11932440 TI - Potassium- or sodium-efflux ATPase, a key enzyme in the evolution of fungi. AB - Potassium is the most abundant cation in cells. Therefore, plant-associated fungi and intracellular parasites are permanently or circumstantially exposed to high K(+) and must avoid excessive K(+) accumulation activating K(+) efflux systems. Because high K(+) and high pH are compatible in natural environments, free-living organisms cannot keep a permanent transmembrane DeltapH and cannot rely only on K(+)/H(+) antiporters, as do mitochondria. This study shows that the Schizosaccharomyces pombe CTA3 is a K(+)-efflux ATPase, and that other fungi are furnished with Na(+)-efflux ATPases, which also pump Na(+). All these fungal ATPases, including those pumping only Na(+), form a phylogenetic group, IID or ENA, among P-type ATPases. By searching in databases and partial cloning of ENA genes in species of Zygomycetes and Basidiomycetes, the authors conclude that probably all fungi have these genes. This study indicates that fungal K(+)- or Na(+)-ATPases evolved from an ancestral K(+)-ATPase, through processes of gene duplication. In yeast hemiascomycetes these duplications have occurred recently and produced bifunctional ATPases, whereas in Neurospora, and probably in other euascomycetes, they occurred earlier in evolution and produced specialized ATPases. In Schizosaccharomyces, adaptation to Na(+) did not involve the duplication of the K(+)-ATPase and thus it retains an enzyme which is probably close to the original one. The parasites Leishmania and Trypanosoma have ATPases phylogenetically related to fungal K(+)-ATPases, which are probably functional homologues of the fungal enzymes. PMID- 11932441 TI - Development of a P1 phagemid system for the delivery of DNA into Gram-negative bacteria. AB - The inability to transform many clinically important Gram-negative bacteria has hampered genetic studies addressing the mechanism of bacterial pathogenesis. This report describes the development and construction of a delivery system utilizing the broad-host-range transducing bacteriophage P1. The phagemids used in this system contain a P1 pac initiation site to package the vector, a P1 lytic replicon to generate concatemeric DNA, a broad-host-range origin of replication and an antibiotic-resistance determinant to select bacterial clones containing the recircularized phagemid. Phagemid DNA was successfully introduced by infection and stably maintained in members of the families Enterobacteriaceae (Escherichia coli, Shigella flexneri, Shigella dysenteriae, Klebsiella pneumoniae and Citrobacter freundii) and Pseudomonadaceae (Pseudomonas aeruginosa). In addition to laboratory strains, these virions were used successfully to deliver phagemids to a number of strains isolated from patients. This ability to deliver genetic information to wild-type strains raises the potential for use in antimicrobial therapies and DNA vaccine development. PMID- 11932443 TI - Physiological consequences associated with overproduction of Mycobacterium tuberculosis FtsZ in mycobacterial hosts. AB - The ftsZ gene of Mycobacterium tuberculosis H37Rv has been characterized as the first step in determining the molecular events involved in the cell division process in mycobacteria. Western analysis revealed that intracellular levels of FtsZ are growth phase dependent in both M. tuberculosis and Mycobacterium smegmatis. Unregulated expression of M. tuberculosis ftsZ from constitutive hsp60 and dnaA promoters in M. tuberculosis hosts resulted in lethality whereas expression from only the hsp60 promoter was toxic in M. smegmatis hosts. Expression of ftsZ from the dnaA promoter in M. smegmatis resulted in approximately sixfold overproduction and the merodiploids exhibited slow growth, an increased tendency to clump and filament, and in some cases produced buds and branches. Many of the cells also contained abnormal and multiple septa. Expression of ftsZ from the chemically inducible acetamidase promoter in M. smegmatis hosts resulted in approximately 22-fold overproduction of FtsZ and produced filamentous cells, many of which lacked any visible septa. Visualization of the M. tuberculosis FtsZ tagged with green fluorescent protein in M. smegmatis by fluorescence microscopy revealed multiple fluorescent FtsZ foci, suggesting that steps subsequent to the formation of organized FtsZ structures but prior to septum formation are blocked in FtsZ-overproducing cells. Together these results suggest that the intracellular concentration of FtsZ protein is critical for productive septum formation in mycobacteria. PMID- 11932442 TI - MabA (FabG1), a Mycobacterium tuberculosis protein involved in the long-chain fatty acid elongation system FAS-II. AB - The fatty acid elongation system FAS-II is involved in the biosynthesis of mycolic acids, which are very long-chain fatty acids of the cell envelope specific to Mycobacterium tuberculosis and other mycobacteria. A potential component of FAS-II, the protein MabA (FabG1), was overexpressed and purified. Sedimentation equilibrium analyses revealed that MabA undergoes a dimer to tetramer self-association with a dissociation constant of 22 microM. The protein was detected by Western blotting in a mycobacterial cell-wall extract that produces mycolic acids and in the FPLC FAS-II fraction. MabA was shown to catalyse the NADPH-specific reduction of beta-ketoacyl derivatives, equivalent to the second step of a FAS-II elongation round. Unlike the known homologous proteins, MabA preferentially metabolizes long-chain substrates (C(8)-C(20)) and has a poor affinity for the C(4) substrate, in agreement with FAS-II specificities. Molecular modelling of MabA structure suggested the presence of an unusually hydrophobic substrate-binding pocket holding a unique Trp residue, suitable for fluorescence spectroscopic analyses. In agreement with the enzyme kinetic data, the spectral properties of MabA were different in the presence of the C(8)-C(16) ligands as compared to the C(4) ligand. Altogether, these data bring out distinctive enzymic and structural properties of MabA, which correlate with its predilection for long-chain substrates, in contrast to most of the other known ketoacyl reductases. PMID- 11932444 TI - Characterization of the genetic locus responsible for the production of ABP-118, a novel bacteriocin produced by the probiotic bacterium Lactobacillus salivarius subsp. salivarius UCC118. AB - ABP-118, a small heat-stable bacteriocin produced by Lactobacillus salivarius subsp. salivarius UCC118, a strain isolated from the ileal-caecal region of the human gastrointestinal tract, was purified to homogeneity. Using reverse genetics, a DNA fragment specifying part of ABP-118 was identified on a 10769 bp chromosomal region. Analysis of this region revealed that ABP-118 was a Class IIb two-peptide bacteriocin composed of Abp118alpha, which exhibited the antimicrobial activity, and Abp118beta, which enhanced the antimicrobial activity. The gene conferring strain UCC118 immunity to the action of ABP-118, abpIM, was identified downstream of the abp118beta gene. Located further downstream of abp118beta, several ORFs were identified whose deduced proteins resembled those of proteins involved in bacteriocin regulation and secretion. Heterologous expression of ABP-118 was achieved in Lactobacillus plantarum, Lactococcus lactis and Bacillus cereus. In addition, the abp118 locus encoded an inducing peptide, AbpIP, which was shown to play a role in the regulation of ABP 118 production. This novel bacteriocin is, to the authors' knowledge, the first to be isolated from a known human probiotic bacterium and to be characterized at the genetic level. PMID- 11932445 TI - Sequence analysis of the lactococcal bacteriophage bIL170: insights into structural proteins and HNH endonucleases in dairy phages. AB - The complete 31754 bp genome of bIL170, a virulent bacteriophage of Lactococcus lactis belonging to the 936 group, was analysed. Sixty-four ORFs were predicted and the function of 16 of them was assigned by significant homology to proteins in databases. Three putative homing endonucleases of the HNH family were found in the early region. An HNH endonuclease with zinc-binding motif was identified in the late cluster, potentially being part of the same functional module as terminase. Three putative structural proteins were analysed in detail and show interesting features among dairy phages. Notably, gpl12 (putative fibre) and gpl20 (putative baseplate protein) of bIL170 are related by at least one of their domains to a number of multi-domain proteins encoded by lactococcal or streptococcal phages. A 110- to 150-aa-long hypervariable domain flanked by two conserved motifs of about 20 aa was identified. The analysis presented here supports the participation of some of these proteins in host-range determination and suggests that specific adsorption to the host may involve a complex multi component system. Divergences in the genome of phages of the 936 group, that may have important biological properties, were noted. Insertions/deletions of units of one or two ORFs were the main source of divergence in the early clusters of the two entirely sequenced phages, bIL170 and sk1. An exchange of fragments probably affected the regions containing the putative origin of replication. It led to the absence in bIL170 of the direct repeats recognized in sk1 and to the presence of different ORFs in the ori region. Shuffling of protein domains affected the endolysin (putative cell-wall binding part), as well as gpl12 and gpl20. PMID- 11932446 TI - Metabolic engineering of lactic acid bacteria, the combined approach: kinetic modelling, metabolic control and experimental analysis. AB - Everyone who has ever tried to radically change metabolic fluxes knows that it is often harder to determine which enzymes have to be modified than it is to actually implement these changes. In the more traditional genetic engineering approaches 'bottle-necks' are pinpointed using qualitative, intuitive approaches, but the alleviation of suspected 'rate-limiting' steps has not often been successful. Here the authors demonstrate that a model of pyruvate distribution in Lactococcus lactis based on enzyme kinetics in combination with metabolic control analysis clearly indicates the key control points in the flux to acetoin and diacetyl, important flavour compounds. The model presented here (available at http://jjj.biochem.sun.ac.za/wcfs.html) showed that the enzymes with the greatest effect on this flux resided outside the acetolactate synthase branch itself. Experiments confirmed the predictions of the model, i.e. knocking out lactate dehydrogenase and overexpressing NADH oxidase increased the flux through the acetolactate synthase branch from 0 to 75% of measured product formation rates. PMID- 11932447 TI - Expression of the Escherichia coli yfiD gene responds to intracellular pH and reduces the accumulation of acidic metabolic end products. AB - The YfiD protein of Escherichia coli has been reported to be an acid-inducible protein. Here it is shown that expression of a yfiD::lac reporter fusion is enhanced up to 3 small middle dot5-fold during acidic growth. The anaerobic transcription factor FNR was confirmed as the major regulator of yfiD expression, and ArcA was found to enhance anaerobic yfiD expression, probably by displacing a repressing FNR dimer in the -93 small middle dot5 region of the promoter. Moreover, the pyruvate sensor PdhR was shown to act as a minor anaerobic repressor of yfiD expression. On the basis of its strong homology to the C terminal region of pyruvate formate-lyase (PFL) it was predicted that YfiD would be a radical-containing enzyme. The YfiD radical was found to be introduced by the PFL-activase enzyme, but unlike PFL, AdhE did not deactivate radicalized YfiD. The extent of radical activation of YfiD was enhanced by low intracellular pH, and thus it was concluded that both yfiD expression and YfiD activity are affected by growth at low pH. The yfiD mutant strain JRG4033 excreted increased levels of organic acids compared to the parental strain when grown in chemostat culture under oxygen-starved conditions, consistent with the acid-inducibility of yfiD expression and the recently reported ability of YfiD to rescue the activity of oxygenolytically cleaved PFL. PMID- 11932448 TI - Escherichia coli aconitases and oxidative stress: post-transcriptional regulation of sodA expression. AB - Escherichia coli possesses two aconitases, a stationary-phase enzyme (AcnA), which is induced by iron and oxidative stress, and a major but less stable enzyme (AcnB), synthesized during exponential growth. In addition to the catalytic activities of the holo-proteins, the apo-proteins function as post transcriptional regulators by site-specific binding to acn mRNAs. Thus, it has been suggested that inactivation of the enzymes could mediate a rapidly reacting post-transcriptional component of the bacterial oxidative stress response. Here it is shown that E. coli acn mutants are hypersensitive to the redox-stress reagents H(2)O(2) and methyl viologen. Proteomic analyses further revealed that the level of superoxide dismutase (SodA) is enhanced in acnB and acnAB mutants, and by exposure to methyl viologen. The amounts of other proteins, including thioredoxin reductase, 2-oxoglutarate dehydrogenase, succinyl-CoA synthetase and chaperone proteins, were also affected in the acn mutants. The altered patterns of sodA expression were confirmed in studies with sodA-lacZ reporter strains. Quantitative Northern blotting indicated that AcnA enhances the stability of the sodA transcript, whereas AcnB lowers its stability. Direct evidence that the apo proteins have positive (AcnA) and negative (AcnB) effects on SodA synthesis was obtained from in vitro transcription-translation experiments. It is suggested that the aconitase proteins of E. coli serve as a protective buffer against the basal level of oxidative stress that accompanies aerobic growth by acting as a sink for reactive oxygen species and by modulating translation of the sodA transcript. PMID- 11932449 TI - Intracellular cyclic AMP concentration is decreased in Salmonella typhimurium fur mutants. AB - It is known that the Fur protein negatively regulates iron-uptake systems in different bacterial species, including Salmonella typhimurium. In this study it has been shown that the intracellular concentration of cyclic AMP (cAMP) is lower in a knockout S. typhimurium fur mutant than in the wild-type strain. According to this, the expression of two cAMP-regulated genes, such as pepE (encoding an alpha-aspartyl dipeptidase) and the Escherichia coli lac operon, is decreased in S. typhimurium fur cells in comparison with wild-type cells. Introduction of an additional mutation in cpdA, encoding a cyclic 3',5'-cAMP phosphodiesterase, recovers wild-type intracellular cAMP concentration in the S. typhimurium fur mutant. Likewise, expression of pepE and the E. coli lac operon was the same in the S. typhimurium fur cpdA double mutant and the wild-type strain. Moreover, these results also demonstrate that the S. typhimurium Fur protein positively regulates the expression of the flhD master operon governing the flagellar regulon. This positive control must be mediated by binding of the S. typhimurium Fur protein to the flhD promoter as indicated by the fact that this promoter tests positive in a Fur titration assay. PMID- 11932450 TI - The Candida albicans Sup35p protein (CaSup35p): function, prion-like behaviour and an associated polyglutamine length polymorphism. AB - The Sup35p protein of Saccharomyces cerevisiae is an essential translation factor whose prion-like properties give rise to the non-Mendelian genetic element [PSI(+)]. In this study the SUP35 gene from the related yeast species Candida albicans has been characterized. The CaSUP35 gene encodes a protein (CaSup35p) of 729 aa which shows 65% amino acid identity to the S. cerevisiae Sup35p protein (ScSup35p), with the C-terminal region showing greater identity (79%) than the N terminal region. The full-length CaSup35p can functionally replace ScSup35p in S. cerevisiae although complementation is only complete when CaSup35p is overexpressed. Complementation only requires expression of the CaSup35p C domain. In S. cerevisiae the full-length CaSup35p is unable to establish a prion-like aggregated state even in the presence of endogenous ScSup35p prion 'seeds', thus confirming the existence of a species barrier in fungal prion propagation. Subcellular localization studies in C. albicans show that although CaSup35p is normally ribosome-associated, when not ribosome-associated, it does not form pelletable high-molecular-mass aggregates characteristic of the ScSup35p in [PSI(+)] strains. Unlike the ScSup35p, the CaSup35p N domain contains a number of polyglutamine repeats although it does contain seven copies of the peptide GGYQQ that is repeated in the ScSup35p N domain. Analysis of the CaSUP35 gene from 14 different strains of C. albicans identified four naturally occurring polymorphisms associated with changes in the length of the largest of the polyglutamine repeats. These findings have important implications for the evolution of fungal prion genes. PMID- 11932451 TI - Homozygosity at the Candida albicans MTL locus associated with azole resistance. AB - Antifungal drug resistance in the pathogenic fungus Candida albicans is a serious threat to the growing population of immunocompromised patients. This study describes a significant correlation between loss of heterozygosity at the C. albicans mating-type-like (MTL) locus and resistance to azole antifungals. A pool of 96 clinical isolates consisting of 50 azole-resistant or susceptible dose dependent isolates and 46 azole-susceptible isolates was screened by PCR for the presence of MTLa1 and MTLalpha1. These genes were used as markers for the MTLa and MTLalpha loci. Both loci were present in 84 of the isolates. Six isolates failed to amplify MTLa1 and six failed to amplify MTLalpha1. Further PCR analysis demonstrated that loss of the MTLa1 and MTLalpha1 genes corresponded to loss of all of the loci-specific genes, resulting in homozygosity at the MTL locus. Southern analysis and single nucleotide polymorphism (SNP) analysis were used to determine that this loss of heterogeneity was due to replacement of one of the MTL loci with a duplicate of the other locus resulting in two homozygous copies of the MTL locus. Of the 12 homozygous isolates, one isolate was sensitive to azole drugs. Statistical analysis of the data demonstrates a strong correlation between homozygosity at the MTL locus and azole resistance (P<0 small middle dot003). In a set of serial isolates, an increase in azole resistance correlated with the loss of heterozygosity at the MTL locus, lending further strength to the correlation. Gene disruptions of the MTL loci were found to have no effect on azole susceptibility. PMID- 11932452 TI - Complications in cell-surface labelling by biotinylation of Candida albicans due to avidin conjugate binding to cell-wall proteins. AB - Initial contact between the opportunistic fungal pathogen Candida albicans and host tissue occurs at the cell surface. Biotin derivatives have been used to label the cell-surface proteins of yeasts, with labelled proteins subsequently detected by avidin-reporter conjugates. Previous work has indicated that avidin can bind to C. albicans proteins in the absence of biotin, suggesting a possible host-cell-recognition mechanism by fungal cell-surface proteins. To investigate this mechanism, Western blots of proteins extracted from biotinylated and mock treated cells were probed with avidin or modified-avidin reagents. Each avidin reagent bound to cell-wall proteins extracted from non-biotinylated cells. Binding did not appear to be due to the lectin-like activity of the cell-wall proteins of C. albicans or to the presence of biotin in the sample itself. Binding was inhibited by added biotin, by the chaotrope KSCN and by NaCl in a concentration-dependent manner, although inhibition varied among the avidin conjugates tested. Thus, the non-specific binding of avidin to the cell-wall proteins of C. albicans appears to involve hydrophobic and electrostatic interactions, depending on the particular avidin species. These observations demonstrate potential pitfalls in the use of avidin-biotin complexes to identify cell-surface molecules and could provide insights into protein-protein interactions at the C. albicans cell wall. PMID- 11932453 TI - Identification of a role for Saccharomyces cerevisiae Cgr1p in pre-rRNA processing and 60S ribosome subunit synthesis. AB - Saccharomyces cerevisiae CGR1 encodes a conserved fungal protein that localizes to the nucleolus. To determine if this localization reflects a role for Cgr1p in ribosome biogenesis two yeast cgr1 mutants were examined for defects in ribosome synthesis: a conditional depletion strain in which CGR1 is under the control of a tetracycline-repressible promoter and a mutant strain in which a C-terminal truncated Cgr1p is expressed. Both strains had impaired growth rates and were hypersensitive to the aminoglycosides paromomycin and hygromycin. Polysome analyses of the mutants revealed increased levels of free 40S subunits relative to 60S subunits, a decrease in 80S monosomes and accumulation of half-mer polysomes. Pulse-chase labelling demonstrated that pre-rRNA processing was defective in the mutants, resulting in accumulation of the 35S, 27S and 7S pre rRNAs and delayed production of the mature 25S and 5 small middle dot8S rRNAs. The synthesis of the 18S and 5S rRNAs was unaffected. Loss of Cgr1 function also caused a partial delocalization of the 5'-ITS1 RNA and the nucleolar protein Nop1p into the nucleoplasm, suggesting that Cgr1p contributes to compartmentalization of nucleolar constituents. Together these findings establish a role for Cgr1p in ribosome biogenesis. PMID- 11932454 TI - Biosynthesis of the dideoxysugar component of jadomycin B: genes in the jad cluster of Streptomyces venezuelae ISP5230 for L-digitoxose assembly and transfer to the angucycline aglycone. AB - Eight additional genes, jadX, O, P, Q, S, T, U and V, in the jad cluster of Streptomyces venezuelae ISP5230, were located immediately downstream of jadN by chromosome walking. Sequence analyses and comparisons implicated them in biosynthesis of the 2,6-dideoxysugar in jadomycin B. The genes were cloned in Escherichia coli, inactivated by inserting an apramycin resistance cassette with a promoter driving transcription of downstream genes, and transferred into Streptomyces venezuelae by intergeneric conjugation. Analysis by HPLC and NMR of intermediates accumulated by cultures of the insertionally inactivated Streptomyces venezuelae mutants indicated that jadO, P, Q, S, T, U and V mediate formation of the dideoxysugar moiety of jadomycin B and its attachment to the aglycone. Based on these results and sequence similarities to genes described in other species producing deoxysugar derivatives, a biosynthetic pathway is proposed in which the jadQ product (glucose-1-phosphate nucleotidyltransferase) activates glucose to its nucleotide diphosphate (NDP) derivative, and the jadT product (a 4,6-dehydratase) converts this to NDP-4-keto-6-deoxy-D-glucose. An NDP hexose 2,3-dehydratase and an oxidoreductase, encoded by jadO and jadP, respectively, catalyse ensuing reactions that produce an NDP-2,6-dideoxy-D-threo 4-hexulose. The product of jadU (NDP-4-keto-2,6-dideoxy-5-epimerase) converts this intermediate to its L-erythro form and the jadV product (NDP-4-keto-2,6 dideoxyhexose 4-ketoreductase) reduces the keto group of the NDP-4-hexulose to give an activated form of the L-digitoxose moiety in jadomycin B. Finally, a glycosyltransferase encoded by jadS transfers the activated sugar to jadomycin aglycone. The function of jadX is unclear; the gene is not essential for jadomycin B biosynthesis, but its presence ensures complete conversion of the aglycone to the glycoside. The deduced amino acid sequence of a 612 bp ORF (jadR*) downstream of the dideoxysugar biosynthesis genes resembles many TetR family transcriptional regulator sequences. PMID- 11932456 TI - Halogenated furanones inhibit quorum sensing through accelerated LuxR turnover. AB - N-acyl-L-homoserine lactones (AHLs) are co-regulatory ligands required for control of the expression of genes encoding virulence traits in many Gram negative bacterial species. Recent studies have indicated that AHLs modulate the cellular concentrations of LuxR-type regulatory proteins by binding and fortifying these proteins against proteolytic degradation (Zhu & Winans, 2001 ). Halogenated furanones produced by the macroalga Delisea pulchra inhibit AHL dependent gene expression. This study assayed for an in vivo interaction between a tritiated halogenated furanone and the LuxR protein of Vibrio fischeri overproduced in Escherichia coli. Whilst a stable interaction between the algal metabolite and the bacterial protein was not found, it was noted by Western analysis that the half-life of the protein is reduced up to 100-fold in the presence of halogenated furanones. This suggests that halogenated furanones modulate LuxR activity but act to destabilize, rather than protect, the AHL dependent transcriptional activator. The furanone-dependent reduction in the cellular concentration of the LuxR protein was associated with a reduction in expression of a plasmid encoded P(luxI)-gfp(ASV) fusion suggesting that the reduction in LuxR concentration is the mechanism by which furanones control expression of AHL-dependent phenotypes. The mode of action by which halogenated furanones reduce cellular concentrations of the LuxR protein remains to be characterized. PMID- 11932455 TI - Nonribosomal biosynthesis of vancomycin-type antibiotics: a heptapeptide backbone and eight peptide synthetase modules. AB - During analysis of the recently identified gene cluster for the glycopeptide antibiotic balhimycin, produced by Amycolatopsis mediterranei DSM 5908, novel genes were identified and characterized in detail. The gene products of four of the identified genes (bpsA, bpsB, bpsC and bpsD) are nonribosomal peptide synthetases (NRPSs); one (Orf1-protein) shows similarities to small proteins associated with several NRPSs without an assigned function. BpsA and BpsB are composed of three modules each (modules 1-6), BpsC of one module (module 7) and BpsD of a minimal module (module 8). Thus, the balhimycin gene cluster encodes eight modules, whereas its biosynthetic product is a heptapeptide. Non-producing mutants were created by a gene disruption of bpsB, an in-frame deletion of bpsC and a gene replacement of bpsD. After establishment of a gene complementation system for Amycolatopsis strains, the replacement mutant of bpsD was complemented, demonstrating for the first time that BpsD, encoding the eighth module, is indeed involved in balhimycin biosynthesis. After feeding with beta hydroxytyrosine the capability of the bpsD mutant to produce balhimycin was restored, demonstrating the participation of BpsD in the biosynthesis of this amino acid. The specificity of four of the eight adenylation domains was determined by ATP/PP(i) exchange assays: modules 4 and 5 activated L-4 hydroxyphenylglycine, module 6 activated beta-hydroxytyrosine and module 7 activated L-3,5-dihydroxyphenylglycine, which is in accordance with the sequence of the non-proteogenic amino acids 4 to 7 of the balhimycin backbone. PMID- 11932458 TI - Efflux of organic acids in Penicillium simplicissimum is an energy-spilling process, adjusting the catabolic carbon flow to the nutrient supply and the activity of catabolic pathways. AB - Continuous cultivation was used to study the effect of glucose, ammonium, nitrate or phosphate limitation on the excretion of tricarboxylic acid (TCA) cycle intermediates by Penicillium simplicissimum. Additionally, the effect of benzoic acid, salicylhydroxamic acid (SHAM) and 2,4-dinitrophenol on TCA cycle intermediates was studied. The physiological state of the fungus was characterized by its glucose and O(2) consumption, its CO(2) production, its intra- and extracellular concentrations of TCA cycle intermediates, as well as by its biomass yield, its maintenance coefficient and its respiratory quotient. The excretion of TCA cycle intermediates was observed during ammonium-, nitrate- and phosphate-limited growth. The highest productivity was found with phosphate limited growth. The respiratory quotient was 1.3 under ammonium limitation and 0.7 under phosphate limitation. Citrate was always the main excreted intermediate. This justifies calling this excretion an energy-spilling process, because citrate excretion avoids the synthesis of too much NADH. The addition of benzoic acid further increased the excretion of TCA cycle intermediates by ammonium-limited hyphae. A SHAM-sensitive respiration was constitutively present during ammonium-limited growth of the fungus. The sum of the excreted organic acids was negatively correlated with the biomass yield (Y(GlcX)). PMID- 11932457 TI - Redundancy, phylogeny and differential expression of Histoplasma capsulatum catalases. AB - Histoplasma capsulatum produces an extracellular catalase termed M antigen, which is similar to catalase B of Aspergillus and Emericella species. Evidence is presented here for two additional catalase isozymes in H. capsulatum. Catalase A is highly similar to a large-subunit catalase in Aspergillus and Emericella species, while catalase P is a small-subunit catalase protein with greatest similarity to known peroxisomal catalases of animals and Saccharomycotina yeasts. Complete cDNAs for the CATA and CATP genes (encoding catalases A and P, respectively) were isolated. The transcriptional expression of the H. capsulatum CATA, CATB (M antigen) and CATP genes was assessed by Northern blot hybridizations on total RNA. Results at the transcript levels for these genes are shown for three conditions: cell morphology (mycelial versus yeast phase cells), oxidative stress (in response to a challenge with H(2)O(2)) and carbon source (glucose vs glycerol). Collectively, these results demonstrated regulation of CATA by both cell morphology and oxidative stress, but not by carbon source, and regulation of CATB and CATP by carbon source but not cell morphology or oxidative stress. A phylogenetic analysis of presently available catalase sequences and intron residences was done. The results support a model for evolution of eukaryotic monofunctional catalase genes from prokaryotic genes. PMID- 11932459 TI - Isolation and biochemical characterization of an endo-1,3-beta-glucanase from Streptomyces sioyaensis containing a C-terminal family 6 carbohydrate-binding module that binds to 1,3-beta-glucan. AB - A gene encoding 1,3-beta-glucanase was isolated from Streptomyces sioyaensis based on an activity plate assay. Analysis of the deduced amino acid sequence of the gene revealed that the matured 1,3-beta-glucanase has two functional domains separated by a stretch of nine glycine residues. The N-terminal domain shares sequence similarity with bacterial endo-1,3-beta-glucanases classified in glycosyl hydrolase family 16 (GHF 16), while the C-terminal domain is a putative carbohydrate-binding module (CBM) grouped into CBM family 6. To characterize the function of each domain, both the full-length and the CBM-truncated versions of the protein were expressed in Escherichia coli and purified to homogeneity. Biochemical data suggest that the glycosyl hydrolase domain preferentially catalyses the hydrolysis of glucans with 1,3-beta linkage, and has an endolytic mode of action. Binding assay indicated that the C-terminal CBM binds to various insoluble beta-glucans (1,3-, 1,3-1,4- and 1,4- linkages) but not to xylan, a primary binding target for most members of CBM family 6. The full-length and the CBM-truncated proteins had similar specific activity (units per mol of hydrolase domain) on soluble 1,3-beta-glucan, whereas the former had much stronger specific activity on insoluble 1,3-beta-glucans, suggesting that the C-terminal CBM enhances the activity of the S. sioyaensis 1,3-beta-glucanase against insoluble substrates, presumably by increasing the frequency of encounter events between the hydrolase domain and the substrate. PMID- 11932460 TI - Influence of extracellular polymeric substances on deposition and redeposition of Pseudomonas aeruginosa to surfaces. AB - In this study, the role of extracellular polymeric substances (EPS) in the initial adhesion of EPS-producing Pseudomonas aeruginosa SG81 and SG81R1, a non EPS-producing strain, to substrata with different hydrophobicity was investigated. The release of EPS by SG81 was concurrent with a decrease in surface tension of a bacterial suspension from 70 to 45 mJ m(-2) that was absent for SG81R1. Both strains adhered faster and in higher numbers to a hydrophilic than to a hydrophobic substratum, but the initial deposition rates and numbers of adhering bacteria in a stationary-end point were highest for the non-EPS producing strain SG81R1, regardless of substratum hydrophobicity. Both strains adhered less to substrata pre-coated with isolated EPS of strain SG81. Furthermore, it was investigated whether bacteria, detached by passing air bubbles, had left behind 'footprints' with an influence on adhesion of newly redepositing bacteria. Redeposition on glass was highest for non-EPS-producing SG81R1 and decreased linearly with the number of times these cycles of detachment and deposition were repeated to become similar to the redeposition of SG81 after six cycles. This indicates that P. aeruginosa SG81 leaves the substratum surface nearly completely covered with EPS after detachment, while SG81R1 releases only minor amounts of surface active EPS, completely covering the substratum after repeated cycles of detachment and adhesion. Atomic force microscopy showed a thick and irregular EPS layer (up to 32 nm) after the first detachment cycle of EPS-producing strain SG81, whereas the putatively non-EPS-producing strain SG81R1 left a 9 nm thin layer after one cycle. X-ray photoelectron spectroscopy indicated that the bacterial footprints consisted of uronic acids, the prevalence of which increased with the number of detachment and deposition cycles. PMID- 11932461 TI - Envelope instability in DNA adenine methylase mutants of Salmonella enterica. AB - Mutants of Salmonella enterica serovar Typhimurium lacking DNA adenine (Dam) methylase show reduced secretion of invasion effectors encoded in the Salmonella pathogenicity island 1 (SPI-1). Concomitant with this alteration, a high number and quantity of extracellular proteins are detected in cultures of Dam(-) mutants. This study shows by subcellular fractionation analysis that the presence of numerous extracellular proteins in cultures of Dam(-) mutants is linked to an exacerbated release of membrane particulate material. The membrane 'leaky' phenotype and the impaired functionality of type III secretion systems were, however, unrelated since exacerbated release of proteins to the medium was evident in Dam(-) strains carrying mutations in either SPI-1 (invA, invJ) or flagellar (flhD) genes. This result supports the view that Dam methylation controls a plethora of cellular processes. Electron microscopy analysis demonstrated that the accumulation of membrane particulate material occurs preferentially as vesicles in stationary cultures of Dam(-) strains. In addition, a reduction in the relative amount of peptidoglycan-associated lipoprotein (PAL), TolB, OmpA and murein lipoprotein (Lpp) bound to peptidoglycan was observed in actively growing Dam(-) mutants. The existence of an envelope defect was further confirmed by the increased sensitivity to deoxycholate exhibited by Dam(-) mutants, mostly during exponential growth. Unexpectedly, lack of Dam methylation neither increased envelope instability nor impaired the association of PAL-Tol Lpp proteins to the peptidoglycan in Escherichia coli. Accordingly, E. coli Dam( ) mutants did not show sensitivity to deoxycholate. Altogether, these results indicate that, besides its role in modulating the secretion of effectors by the SPI-1-encoded type III apparatus, Dam methylation controls cell envelope integrity in S. enterica. PMID- 11932463 TI - Interaction of human Tamm-Horsfall glycoprotein with Bordetella pertussis toxin. AB - Tamm-Horsfall glycoprotein (THP), which is synthesized by renal tubular cells, is the most abundant protein in normal human urine. Although its physiological function remains unclear, it has been proposed that THP may act as a defence factor against urinary tract infections by inhibiting the binding of S- and P fimbriated Escherichia coli to renal epithelial cells. Because THP-related proteins are also found in the superficial layers of the oral mucosa, the authors investigated the ability of THP to interfere with the cytoadherence of pathogenic bacteria that colonize mucosal surfaces other than those of the urogenital tract. In this report, it is shown that THP binds to virulent Bordetella pertussis and reduces its adherence to both renal and pulmonary epithelial cells. This cytoadherence inhibitory effect was not observed with a B. pertussis mutant lacking the pertussis toxin (PTX) operon, and was dependent on the direct interaction of THP with the S2 subunit within the PTX B oligomer. The authors also show that the glycosylation moiety of THP is crucial for its binding to PTX. The THP-PTX interaction was exploited to develop an affinity chromatography method that allows a one-step purification of active PTX. These observations suggest that besides its anti-adherence activity, THP may also trap toxins produced by pathogenic bacteria that colonize mucosal surfaces. PMID- 11932462 TI - Construction and characterization of a nonpigmented mutant of Porphyromonas gingivalis: cell surface polysaccharide as an anchorage for gingipains. AB - A nonpigmented mutant of Porphyromonas gingivalis was constructed by using transposon mutagenesis. The mutant possessed the transposon DNA at the novel gene porR. Gene targeted mutagenesis revealed that porR was responsible for pigmentation. The porR gene shared similarities with genes of the degT family, the products of which are now considered to be transaminases involved in biosynthesis of sugar portions of cell-surface polysaccharides and aminoglycosides. The porR mutant showed a pleiotropic phenotype: delayed maturation of fimbrillin, preferential presence of Rgp and Kgp proteinases in culture supernatants, and no haemagglutination. The porR mutant had altered phenol extractable polysaccharide compared to the porR(+) sibling strain. A mAb, 1B5, that reacts with sugar portions of P. gingivalis cell surface polysaccharide and membrane-type Rgp proteinase showed no reaction with the cell lysates of the porR mutant. These results indicate that porR is involved in biosynthesis of cell surface polysaccharide that may function as an anchorage for Rgp, Kgp, haemagglutinins and the haemoglobin receptor protein. PMID- 11932464 TI - Identification of genetic differences between two Campylobacter jejuni strains with different colonization potentials. AB - The consumption of poultry meat contaminated with Campylobacter jejuni is considered to be a risk factor for human campylobacteriosis. The development of targeted strategies to control campylobacters in broilers would benefit from knowledge of those bacterial factors important in colonization of the avian gut. During preliminary studies it was noted that C. jejuni NCTC 11168 was a poorer colonizer of chickens than strain 81116. This poor colonization could not be fully restored by in vivo passage, suggesting that it was a genetically endowed property of strain 11168. As the genome sequence is available for this strain, the technique of subtractive hybridization was used to identify gene fragments of strain 81116 not present in strain 11168. After two screening cycles, 24 out of 42 clones were identified as having DNA inserts specific for strain 81116. Six of these 24 clones contained gene fragment inserts with similarities to restriction modification enzymes found in other bacteria. Two inserts had similarity to arsenic-resistance genes, whereas four others had similarities to cytochrome c oxidase III, dTDP-glucose 4,6-dehydratase, gamma-glutamyl transpeptidase and an abortive phage-resistance protein. At least some of these genes may be involved with colonization. A further six inserts had weak similarities to hypothetical proteins or to proteins with assigned functions from strain 11168. The remaining six clones had gene-fragment inserts with no database matches. Southern-blot analysis confirmed that strain-dependent variation existed for each of these DNA inserts. These results indicate that subtractive hybridization can successfully identify genes that are absent from the only C. jejuni strain for which the genome sequence is currently available. PMID- 11932465 TI - The benPK operon, proposed to play a role in transport, is part of a regulon for benzoate catabolism in Acinetobacter sp. strain ADP1. AB - BenM and CatM are distinct, but similar, LysR-type transcriptional regulators of the soil bacterium Acinetobacter sp. strain ADP1. Together, the two regulators control the expression of at least 14 genes involved in the degradation of aromatic compounds via the catechol branch of the beta-ketoadipate pathway. In these studies, BenM and CatM were each purified to homogeneity to test the possibility that they regulate the expression of two additional genes, benP and benK, that are adjacent to benM on the chromosome. Each regulator bound to a DNA fragment containing the benP promoter region. Additional transcriptional studies suggested that benP and benK are co-transcribed as an operon, and a site of transcription initiation was identified. Alignment of this initiation site with those of several CatM- and BenM-regulated genes revealed common regulatory motifs. Mutants lacking both CatM and BenM failed to activate benP transcription. The ability of each protein to regulate gene expression was inferred from strains lacking either CatM or BenM that were still capable of increasing benP expression in response to cis,cis-muconate. This compound has previously been shown to induce all enzymes of the catechol branch of the beta-ketoadipate pathway through a complex transcriptional circuit involving CatM and BenM. Thus, the regulated expression of the benPK operon in concert with other genes of the regulon is consistent with the model that BenP, a putative outer-membrane porin, and BenK, an inner-membrane permease, transport aromatic compounds in strain ADP1. PMID- 11932466 TI - Difference in substrate specificity divides the yeast alkali-metal-cation/H(+) antiporters into two subfamilies. AB - Yeast plasma membrane Na(+)/H(+) antiporters (TC 2.A.36) share a high degree of similarity at the protein level. Expression of four antiporters (Saccharomyces cerevisiae Nha1p, Candida albicans Cnh1p, Zygosaccharomyces rouxii ZrSod2-22p and Schizosaccharomyces pombe sod2p) in a SACCH: cerevisiae mutant strain lacking both Na(+)-ATPase and Na(+)/H(+) antiporter genes made it possible to study the transport properties and contribution to cell salt tolerance of all antiporters under the same conditions. The ZrSod2-22p of the osmotolerant yeast Z. rouxii has the highest transport capacity for lithium and sodium but, like the SCHIZ: pombe sod2p, it does not recognize K(+) and Rb(+) as substrates. The SACCH: cerevisiae Nha1p and C. albicans Cnh1p have a broad substrate specificity for at least four alkali metal cations (Na(+), Li(+), K(+), Rb(+)), but their contribution to overall cell tolerance to high external concentration of toxic Na(+) and Li(+) cations seems to be lower compared to the antiporters of SCHIZ: pombe and especially Z. rouxii. PMID- 11932467 TI - Polymorphism in repeated 16S rRNA genes is a common property of type strains and environmental isolates of the genus Vibrio. AB - Analysis of the 16S rDNAs obtained from cultures of single colonies of either type collection strains or environmental strains of the genus Vibrio revealed the presence of polymorphism in every one of the strains examined. Polymorphism was detected by visualization of heteroduplexes produced after 16S rDNA PCR amplification, a procedure that allows for the screening of a large number of isolates. Amplified 16S rDNAs obtained from both Vibrio parahaemolyticus and an environmental strain were cloned. Their nucleotide sequences revealed differences of up to 2% among 16S rDNAs from the same strain. Polymorphic sites were concentrated in a recognized variable stem-loop of bacterial 16S rRNA that contained in some cases up to 83% of the total mismatches observed. Most of the substitutions present in the stem-loop region showed compensating base covariation. The accumulation of so many compensating changes in the stem-loop region implies that the divergence of the different versions of this stem-loop is relatively ancient. This divergence could be the result of either a selection process or a lateral transfer of independently evolved genes. PMID- 11932468 TI - Images in clinical medicine. Porcelain gallbladder. PMID- 11932469 TI - The ethics of partial-liver donation. PMID- 11932470 TI - Screening of infants and mortality due to neuroblastoma. AB - BACKGROUND: Neuroblastoma, the most common extracranial solid tumor that occurs in early childhood, can be identified in the preclinical stages by the detection of catecholamines in the urine. However, it is unknown whether routine screening for neuroblastoma reduces mortality due to this disease. METHODS: Through their parents, we offered screening for neuroblastoma at three weeks and six months of age to all 476,654 children born in the province of Quebec, Canada, during a five year period (May 1, 1989, through April 30, 1994). The participation rate was 92 percent. The rate of death due to neuroblastoma was determined and compared with the rates in several unscreened control populations born during the same period. RESULTS: Among children younger than eight years of age in the Quebec cohort, there were 22 deaths due to neuroblastoma; the cumulative (+/-SE) mortality rate due to neuroblastoma was 4.78+/-1.14 per 100,000 children over a period of nine years. The standardized incidence ratios for death due to neuroblastoma for the Quebec cohort were 1.11 (95 percent confidence interval, 0.64 to 1.92) as compared with a control group in Ontario, Canada; 0.90 (95 percent confidence interval, 0.48 to 1.70) as compared with a control group in Minnesota; 1.40 (95 percent confidence interval, 0.81 to 2.41) as compared with a control group in Florida; and 0.96 (95 percent confidence interval, 0.56 to 1.66) as compared with a control group in the Greater Delaware Valley. The standardized mortality ratio for the Quebec cohort as compared with the rest of Canada was 1.39 (95 percent confidence interval, 0.85 to 2.30); the odds ratio for the comparison with a cohort born in Quebec before the screening program began was 0.98 (95 percent confidence interval, 0.54 to 1.77). CONCLUSIONS: Screening infants for neuroblastoma does not appear to reduce mortality due to this disease. PMID- 11932471 TI - Neuroblastoma screening at one year of age. AB - BACKGROUND: Neuroblastoma is the second most common type of childhood tumor. It is not known whether screening for neuroblastoma at one year of age reduces the incidence of metastatic disease or mortality due to neuroblastoma. METHODS: We offered urine screening for neuroblastoma at approximately one year of age to 2,581,188 children in 6 of 16 German states from 1995 to 2000. A total of 2,117,600 eligible children in the remaining states served as controls. We compared the two groups in terms of the incidence of disseminated disease and mortality from neuroblastoma. RESULTS: A total of 1,475,773 children (61.2 percent of those who were born between July 1, 1994, and October 31, 1999) underwent screening. In this group, neuroblastoma was detected by screening in 149 children, of whom 3 have died. Fifty-five children who had negative screening tests were subsequently given a diagnosis of neuroblastoma; 14 of these children have died. The screened group and children in the control area had a similar incidence of stage 4 neuroblastoma (3.7 cases per 100,000 screened children [95 percent confidence interval, 2.7 to 4.7] and 3.8 per 100,000 controls [95 percent confidence interval, 2.9 to 4.6]) and a similar rate of death among children with neuroblastoma (1.3 deaths per 100,000 screened children [95 percent confidence interval, 0.7 to 1.8] and 1.2 per 100,000 controls [95 percent confidence interval, 0.7 to 1.7]). Comparison of the screened group and the children in the control area revealed substantial overdiagnosis in the former group (an estimated rate of 7 cases per 100,000 children [95 percent confidence interval, 4.6 to 9.2]); the overdiagnosis rate represents children who had neuroblastoma that was diagnosed by screening but who would not benefit from earlier diagnosis and treatment. CONCLUSIONS: The present findings do not support the usefulness of general screening for neuroblastoma at one year of age. PMID- 11932472 TI - Primary chemoprevention of familial adenomatous polyposis with sulindac. AB - BACKGROUND: Familial adenomatous polyposis is caused by a germ-line mutation in the adenomatous polyposis coli gene and is characterized by the development of hundreds of colorectal adenomas and, eventually, colorectal cancer. Nonsteroidal antiinflammatory drugs can cause regression of adenomas, but whether they can prevent adenomas is unknown. METHODS: We conducted a randomized, double-blind, placebo-controlled study of 41 young subjects (age range, 8 to 25 years) who were genotypically affected with familial adenomatous polyposis but phenotypically unaffected. The subjects received either 75 or 150 mg of sulindac orally twice a day or identical-appearing placebo tablets for 48 months. The number and size of new adenomas and side effects of therapy were evaluated every four months for four years, and the levels of five major prostaglandins were serially measured in biopsy specimens of normal-appearing colorectal mucosa. RESULTS: After four years of treatment, the average rate of compliance exceeded 76 percent in the sulindac group, and mucosal prostaglandin levels were lower in this group than in the placebo group. During the course of the study, adenomas developed in 9 of 21 subjects (43 percent) in the sulindac group and 11 of 20 subjects in the placebo group (55 percent) (P=0.54). There were no significant differences in the mean number (P=0.69) or size (P=0.17) of polyps between the groups. Sulindac did not slow the development of adenomas, according to an evaluation involving linear longitudinal methods. CONCLUSIONS: Standard doses of sulindac did not prevent the development of adenomas in subjects with familial adenomatous polyposis. PMID- 11932473 TI - Images in clinical medicine. Hematogenous anaerobic osteomyelitis. PMID- 11932474 TI - Understanding the treatment preferences of seriously ill patients. AB - BACKGROUND: The questions patients are asked about their preferences with regard to life-sustaining treatment usually focus on specific interventions, but the outcomes of treatment and their likelihood affect patients' preferences. METHODS: We administered a questionnaire about treatment preferences to 226 persons who were 60 years of age or older and who had a limited life expectancy due to cancer, congestive heart failure, or chronic obstructive pulmonary disease. The study participants were asked whether they would want to receive a given treatment, first when the outcome was known with certainty and then with different likelihoods of an adverse outcome. The outcome without treatment was specified as death from the underlying disease. RESULTS: The burden of treatment (i.e., the length of the hospital stay, extent of testing, and invasiveness of interventions), the outcome, and the likelihood of the outcome all influenced treatment preferences. For a low-burden treatment with the restoration of current health, 98.7 percent of participants said they would choose to receive the treatment (rather than not receive it and die), but 11.2 percent of these participants would not choose the treatment if it had a high burden. If the outcome was survival but with severe functional impairment or cognitive impairment, 74.4 percent and 88.8 percent of these participants, respectively, would not choose treatment. The number of participants who said they would choose treatment declined as the likelihood of an adverse outcome increased, with fewer participants choosing treatment when the possible outcome was functional or cognitive impairment than when it was death. Preferences did not differ according to the primary diagnosis. CONCLUSIONS: Advance care planning should take into account patients' attitudes toward the burden of treatment, the possible outcomes, and their likelihood. The likelihood of adverse functional and cognitive outcomes of treatment requires explicit consideration. PMID- 11932475 TI - Responding to medical events during commercial airline flights. PMID- 11932476 TI - Adult-to-adult transplantation of the right hepatic lobe from a living donor. PMID- 11932477 TI - The science and politics of screening newborns. PMID- 11932478 TI - Cyclooxygenase inhibition in cancer--a blind alley or a new therapeutic reality? PMID- 11932479 TI - Autonomy reconsidered. PMID- 11932480 TI - Treatment of acute hepatitis C with interferon alfa-2b. PMID- 11932481 TI - Antioxidant vitamins and coronary disease. PMID- 11932482 TI - B vitamins and restenosis after coronary angioplasty. PMID- 11932483 TI - Nephrectomy for metastatic renal-cell cancer. PMID- 11932484 TI - Lowering cardiac risk in noncardiac surgery. PMID- 11932485 TI - Hidden barriers to improvement in the quality of health care. PMID- 11932486 TI - Blood substitute and erythropoietin therapy in a severely injured Jehovah's witness. PMID- 11932487 TI - Design of ET(B) receptor agonists: NMR spectroscopic and conformational studies of ET7-21[Leu7, Aib11, Cys(Acm)15]. AB - In a previous report we have shown that the endothelin-B receptor-selective linear endothelin peptide, ET-1[Cys (Acm)1,15, Ala3, Leu7, Aib11], folds into an alpha-helical conformation in a methanol-d3/water co-solvent [Hewage et al. (1998) FEBS Lett., 425, 234-238]. To study the requirements for the structure activity relationships, truncated analogues of this peptide were subjected to further studies. Here we report the solution conformation of ET7-21[Leu7, Aib11, Cys(Acm)15], in a methanol-d3/water co-solvent at pH 3.6, by NMR spectroscopic and molecular modelling studies. Further truncation of this short peptide results in it displaying poor agonist activity. The modelled structure shows that the peptide folds into an alpha-helical conformation between residues Lys9-His16, whereas the C-terminus prefers no fixed conformation. This truncated linear endothelin analogue is pivotal for designing endothelin-B receptor agonists. PMID- 11932488 TI - Knowledge-based selection of targets for structural genomics. AB - The problem of rational target selection for protein structure determination in structural genomics projects on microbes is addressed. A flexible computational procedure is described that directly incorporates the whole body of annotation available in the PEDANT genome database into the sequence clustering and selection process in order to identify proteins that are likely to possess currently unknown structural domains. Filtering out gene products based on predicted structural features, such as known three-dimensional structures and transmembrane regions, allows one to reduce the complexity of neighbor relationships between sequences and all but eliminates the need for further partitioning of single-linkage clusters into disjoint protein groups corresponding to homologous families. The results of a large-scale computation experiment in which exemplary target selection for 32 prokaryotic genomes was conducted are presented. PMID- 11932489 TI - Molecular dynamics simulations as a tool for improving protein stability. AB - Haloalkane dehalogenase (DhlA) was used as a model protein to explore the possibility to use molecular dynamics (MD) simulations as a tool to identify flexible regions in proteins that can serve as a target for stability enhancement by introduction of a disulfide bond. DhlA consists of two domains: an alpha/beta hydrolase fold main domain and a cap domain composed of five alpha-helices. MD simulations of DhlA showed high mobility in a helix-loop-helix region in the cap domain, involving residues 184-211. A disulfide cross-link was engineered between residue 201 of this flexible region and residue 16 of the main domain. The mutant enzyme showed substantial changes in both thermal and urea denaturation. The oxidized form of the mutant enzyme showed an increase of the apparent transition temperature from 47.5 to 52.5 degrees C, whereas the T(m,app) of the reduced mutant decreased by more than 8 degrees C compared to the wild-type enzyme. Urea denaturation results showed a similar trend. Measurement of the kinetic stability showed that the introduction of the disulfide bond caused a decrease in activation free energy of unfolding of 0.43 kcal mol(-1) compared to the wild type enzyme and also indicated that the helix-loop-helix region was involved early in the unfolding process. The results show that MD simulations are capable of identifying mobile protein domains that can successfully be used as a target for stability enhancement by the introduction of a disulfide cross-link. PMID- 11932490 TI - Protein sequence comparison based on the wavelet transform approach. AB - A protein's chemical properties, the chain conformation, the function of the protein and its species specificity are determined by the information contained in the amino acid sequence. Proteins of similar functions have at some level sequential identical amino acid sequences. The closer the phylogenetic relationship, the more similar are the sequences. To find the similarities between two or more protein sequences is of great importance for protein sequence analysis. The differences in the amino acid sequences permit the construction of a family tree of evolution. In this work, a comparison method was devised that is capable of analysing a protein sequence 'hierarchically', i.e. it can examine a protein sequence at different spatial resolutions. Based on a wavelet decomposition of protein sequences and a cross-correlation study, a sequence scale similarity concept is proposed for generating a similarity vector, which renders the comparison of two sequences feasible at different spatial resolutions (scales). This new similarity concept is an expansion of the conventional sequence similarity, which only takes into account the local pairwise amino acid match and ignores the information contained in coarser spatial resolutions. PMID- 11932491 TI - Analysis of the human hephaestin gene and protein: comparative modelling of the N terminus ecto-domain based upon ceruloplasmin. AB - Hephaestin was implicated in mammalian iron homeostasis following its identification as the defective gene in murine sex-linked anaemia. It is a member of the family of copper oxidases that includes mammalian ceruloplasmin, factors V and VIII, yeast fet3 and fet5 and bacterial ascorbate oxidase. Hephaestin is different from ceruloplasmin, a soluble ferroxidase, in having a membrane spanning region towards the C-terminus. Here we report the gene structure, spanning approximately 100 kb, of the human homologue of mouse hephaestin. The sequence was assembled from the cDNA clones and the chromosome X genomic sequence data available at the Sanger Centre. It has an open reading frame that encodes a protein of 1158 residues, 85% identical with the murine homologue. A model of the N-terminal ecto-domain has been built based on the known three-dimensional structure of human ceruloplasmin. The overall tertiary structure for the hephaestin and the putative residues involved in binding copper and iron appear to be highly conserved between these proteins, which suggests they share the same fold and a conserved function. PMID- 11932492 TI - A low-resolution 3D model of the tetrameric alcohol dehydrogenase from Sulfolobus solfataricus. AB - We describe the computation of a model of the thermophilic NAD-dependent homotetrameric alcohol dehydrogenase from the archaeon Sulfolobus solfataricus (SsADH). Modeling is based on the knowledge that each monomer contains two Zn ions with catalytic and structural function, respectively. In the database of known structures, proteins with similar functions are either dimers containing two zinc ions per monomer or tetramers with one zinc ion per monomer. In any case, the sequence identity of the target to the possible templates is low. A threading procedure is therefore developed which includes constraints taking into account residue conservation both at the zinc ion binding and at the monomer monomer interaction sites in the tetrameric unit. The model is consistent with previously reported data. Furthermore, cross-linking experiments are described which support the computed tetrameric model. PMID- 11932493 TI - Impact of single-residue mutations on the structure and function of ovispirin/novispirin antimicrobial peptides. AB - We studied three model antibacterial peptides that resembled the N-terminal 18 amino acids of SMAP-29, an alpha-helical, antimicrobial peptide of sheep. Although the parent compound, ovispirin-1 (KNLRR IIRKI IHIIK KYG), was potently antimicrobial, it was also highly cytotoxic to human epithelial cells and hemolytic for human erythrocytes. Single residue substitutions to ovispirin-1 yielded two substantially less cytotoxic peptides (novispirins), with intact antimicrobial properties. One of these, novispirin G-10, differed from ovispirin 1 only by containing glycine at position 10, instead of isoleucine. The other, novispirin T-7, contained threonine instead of isoleucine at position 7. We determined the three-dimensional solution structures of all three peptides by circular dichroism spectroscopy and two-dimensional nuclear magnetic resonance spectroscopy. Although all retained an amphipathic helical structure in 2,2,2 trifluoroethanol, they manifested subtle fine-structural changes that evidently impacted their activities greatly. These findings show that simple structural modifications can 'fine-tune' an antimicrobial peptide to minimize unwanted cytotoxicity while retaining its desired activity. PMID- 11932494 TI - Mapping of a hapten-binding site: molecular modeling and site-directed mutagenesis study of an anti-atrazine antibody. AB - A three-dimensional model of the variable domain of the atrazine-specific Fab fragment K411B was constructed by molecular modeling using known structures of highly homologous immunoglobulins as templates. Molecular dynamic simulations and cross-reactivity data were used to predict residues responsible for the binding of the hapten 4-chloro-6-(isopropylamino)-1,3,5-triazine-2-(6 aminohexanecarboxylic acid) (iPr/Cl/C6) instead of atrazine. Specific binding pockets could be defined for the chlorine, the isopropylamino group and the C6 spacer of the hapten. The influence of various amino acids on hapten binding was investigated by site-directed mutagenesis, and the effect of these mutations was analyzed by capture ELISA using the hapten iPr/Cl/C6 and 4-amino-6-chloro-1,3,5 triazine-2-(6-aminohexanecarboxylic acid) (H/Cl/C6). GlyH100a seems to be important in determining the conformation of the heavy-chain complementarity determining region H3; replacing it with any other residue prevented the binding of the hapten. Altering residues responsible for the binding of the chlorine atom (TrpH33, GluH50 and TyrL96) decreased the affinity significantly. Hapten-spacer recognition can be attributed to the interaction with PheL32; replacing PheL32 by leucine reduced the affinity towards iPr/Cl/C6. A triple mutant Fab fragment (GlnL89Glu, ValH37Ile and GluL3Val) showed an affinity 5-fold greater towards iPr/Cl/C6 compared to the wild-type K411B, as a result of better recognition of the isopropylamino group of iPr/Cl/C6. PMID- 11932496 TI - Alternative CUG codon usage (Ser for Leu) in Pichia farinosa and the effect of a mutated killer gene in Saccharomyces cerevisiae. AB - The halotolerant yeast Pichia farinosa KK1 strain produces a killer toxin termed SMKT (salt-mediated killer toxin). Mass spectrometry and Edman sequencing of peptides from the mature SMKT and secreted protoxin demonstrate that positions specified by the CUG codon contain unmodified serine (Ser) in P.farinosa. In order to express the authentic SMK1 product in Saccharomyces cerevisiae, which uses the universal genetic code, the three CUG codons corresponding to Ser87, Ser137 and Ser206 in the SMK1 gene were changed to universal Ser codons by site directed mutagenesis. The expression of the modified SMK1 gene with universal Ser codons was lethal in S.cerevisiae, as well as that of the unmodified SMK1 gene with the CUG codons. The secretion of protoxin with the authentic amino acid sequence from the modified SMK1 was significantly increased, whereas the transcription level of SMK1 was not affected in the presence or absence of CUG codon. Our results provide the first in vivo evidence that non-universal decoding of CUG is used in a hemiascomycetous yeast, P.farinosa. PMID- 11932495 TI - A recombinant bacterial cell surface (S-layer)-major birch pollen allergen-fusion protein (rSbsC/Bet v1) maintains the ability to self-assemble into regularly structured monomolecular lattices and the functionality of the allergen. AB - The mature crystalline bacterial cell surface (S-layer) protein SbsC of Bacillus stearothermophilus ATCC 12980 comprises amino acids 31-1099 and assembles into an oblique lattice type. As the deletion of up to 179 C-terminal amino acids did not interfere with the self-assembly properties of SbsC, the sequence encoding the major birch pollen allergen (Bet v1) was fused to the sequence encoding the truncated form rSbsC(31-920). The S-layer fusion protein, termed rSbsC/Bet v1, maintained the ability to self-assemble into flat sheets and open-ended cylinders. The presence and the functionality of the fused Bet v1 sequence was proved by blot experiments using BIP1, a monoclonal antibody against Bet v1 and Bet v1-specific IgE-containing serum samples from birch pollen allergic patients. The location and accessibility of the allergen moiety on the outer surface of the S-layer lattice were demonstrated by immunogold labeling of the rSbsC/Bet v1 monolayer, which was obtained by oriented recrystallization of the S-layer fusion protein on native cell wall sacculi. Thereby, the specific interactions between the N-terminal part of SbsC and a distinct type of secondary cell wall polymer were exploited. This is the first S-layer fusion protein described that had retained the specific properties of the S-layer protein moiety in addition to those of the fused functional peptide sequence. PMID- 11932497 TI - AS madness. PMID- 11932498 TI - Teaching aids for developing countries. PMID- 11932499 TI - The Library's new strategy. PMID- 11932500 TI - Mosquitoes and tyres. AB - Car tyres not only help people to move about, they also assist exotic mosquitoes in travelling thousands of miles around the world to colonise new areas. The remarkable adaptability of mosquitoes that has led to their enormous success in the past can be seen close to home. PMID- 11932501 TI - Coral gardener of the Maldives. AB - Abdul Azeez Abdul Hakeem is a man with a vision. He is 53 years old, with a degree in agronomy and an idea, if not an obsession: to nurse and propagate corals on a large scale. His motive, however, is not to provide for the international marine aquarium trade but as a safeguard for the livelihood of his country, the Maldives. PMID- 11932502 TI - Proteomics: the protein revolution. AB - Proteomics, the latest scientific buzzword and research field, represents a milestone in biological research, as proteins return to centre stage. But what has led to this protein renaissance and why has proteomics become a key research tool in the post-genomic era? The answers to these questions lie in the huge progress that has been made in the area of genomics. PMID- 11932503 TI - Influenza A: a threatening virus with two faces. AB - In 1918, a great 'forgotten plague' wiped out fifty million people around the world - many of them young and healthy. The killer disease was flu. What was it that made this outbreak so deadly and would we be prepared if another pandemic struck? PMID- 11932504 TI - How plants see themselves--self-incompatibility in flowering plants. AB - Plants have been propagating themselves by cloning for millennia. It is, however, widely recognised that mixing genes with other individuals of the same species makes better evolutionary sense, as it provides the variation that is the raw material for natural selection. How, then, do some plants prevent self fertilisation? PMID- 11932505 TI - Painting by numbers: genetic diseases and chromosome paints. AB - The image above comes from a new technique, known as chromosome painting. This beautiful snapshot of the central molecule of life - DNA - in its natural form allows us to learn about chromosomes and nuclear structures. It also allows the fast diagnosis of chromosome abnormalities. PMID- 11932506 TI - A compromised participation? AB - Agriculture in Britain has become the focus of international attention in recent years. Many have questioned the changes that have occurred in the industry, and the typical assumption is that agriculture has become increasingly unsustainable and remote form consumers. The contribution of agricultural research to all of this is also being dissected: after all, who is supposed to benefit? PMID- 11932507 TI - Who was... John Henri Fabre? AB - The books of Jean Henri Fabre, replete with lively accounts of his observations on nature, inspired generations of children from all over the world. The detail in his study of insects and the entertaining presentation allowed readers to absorb his fascination. Yet, he was a physics teacher by profession and virtually self taught on matters of entomology. PMID- 11932508 TI - What's in a name? AB - Have you ever wondered why professionals have so many complicated names for things? Two reasons come to mind: efficient communication, but also 'show'. Connecting with a wider audience means learning to tread the fine line between perfect accuracy and being understood. PMID- 11932509 TI - Comment on mu(1,3)-azido-diazidotetrakis(1,10-phenanthroline)dicopper(II) azide tetrahydrate. AB - The supramolecular structure of the title dimeric azido complex of copper(II), [Cu(2)(mu(1,3)-N(3))(N(3))(2)(phen)(4)](N(3)) x 4H(2)O (phen is 1,10 phenanthroline, C(12)H(8)N(2)) [Cheng, Hu, Wang & Ye (2002). Acta Cryst. C58, m12 m13], which was originally described in terms of hydrogen-bonded chains, has been re-interpreted as two-dimensional hydrogen-bonded sheets built from R(6)4(12) and R(10)10(28) rings, taking into account the complete hydogen-bonding pattern. PMID- 11932511 TI - Tetrapotassium nonastannide, K(4)Sn(9). AB - In K(4)Sn(9), which crystallizes with a new structure type, the Sn atoms form isolated Wade nido-[Sn(9)]4- clusters of approximate C(4v) symmetry (monocapped square antiprisms), with Sn-Sn distances ranging from 2.9264 (9) to 3.348 (1) A. The cluster anions are separated by K+ cations and are in a hexagonal close packed arrangement. PMID- 11932512 TI - Hg(3)Se(3)O(10), a mercury(II) compound with mixed-valence oxoselenium(IV/VI) anions. AB - The title compound, trimercury(II) bis[selenite(IV)] selenate(VI), contains three crystallographically inequivalent HgII cations with coordination numbers of eight (denoted Hg1 and Hg2) and five (denoted Hg3). The corresponding coordination polyhedra around the metal atoms might be described as intermediates between a square antiprism and a triangulated dodecahedron for both Hg1 and Hg2, and a strongly distorted truncated octahedron for Hg3. (infinity)2[HgO(8/2)] layers of edge-sharing [HgO(8)] polyhedra propagate parallel to the bc plane, and are connected via SeVIO(4) tetrahedra and [Hg3O(5)] polyhedra along the a axis, forming an arrangement with channels propagating parallel to the b axis. The two independent SeIVO(3) pyramids bridge the Hg atoms, and the non-bonding orbitals of the SeIV ions protrude into the channels from opposite sides. PMID- 11932513 TI - Ba(14)Cu(2)In(4)N(7), a new subnitride with isolated nitridocuprate groups and indium clusters. AB - Single crystals of Ba(14)Cu(2)In(4)N(7), tetradecabarium dicopper tetraindium heptanitride, were synthesized by slow cooling from 1023 K at 7 MPa of N(2) using an Na flux. The compound crystallizes in the monoclinic space group P2/m with Z = 2, and contains 0[CuN(2)] nitridocuprate units and distorted 0[In(4)] clusters. One Ba atom, not connected to any N atoms, is surrounded by 12 other Ba atoms in a barium cuboctahedron. The structural formula is expressed as (Ba)Ba(27)N(6)[CuN(2)](4)[In(4)](2). PMID- 11932514 TI - catena-Poly[[triphenyltin(IV)]-mu-hydroxo-kappa2O:O] at 120 K. AB - The structure of the title compound, [Sn(C(6)H(5))(3)(OH)](n), has been re investigated at 120 (2) K. The hydroxyl H atom was readily located and the threefold coordination about the O atom is planar. There are no hydrogen bonds involving the hydroxyl group, either as donor or as acceptor. PMID- 11932515 TI - Triaquatris(2,6-dinitrophenolato-kappa2O1,O2)yttrium(III). AB - In the title compound, [Y(C(6)H(3)N(2)O(5))(3)(H(2)O)(3)], the Y atom is nine coordinate with a slightly distorted tricapped trigonal prismatic coordination geometry. The Y(III) ion is coordinated to three bidentate 2,6-dinitrophenolate ligands and three water molecules. The Y-O bond distances are in the range 2.217 (3)-2.754 (4) A, with the Y-O distances from the nitro groups being longer than those from the water molecules and the phenol groups. The coordinated NO(2) groups are almost coplanar with the benzene rings. PMID- 11932516 TI - trans-Bis(cyano-kappaC)(1,4,8,11-tetraazacyclotetradecane-kappa4N)manganese(III) perchlorate, a low-spin manganese(III) complex. AB - The crystal structure of the low-spin (S = 1) MnIII complex [Mn(CN)(2)(C(10)H(24)N(4))]ClO(4), or trans-[Mn(CN)(2)(cyclam)](ClO(4)) (cyclam is the tetradentate amine ligand 1,4,8,11-tetraazacyclotetradecane), is reported. The structural parameters in the Mn(cyclam) moiety are found to be insensitive to both the spin and the oxidation state of the Mn ion. The difference between high- and low-spin Mn(III) complexes is that a pronounced tetragonal elongation of the coordination octahedron occurs in high-spin complexes and a slight tetragonal compression is seen in low-spin complexes, as in the title complex. PMID- 11932517 TI - (2,2'-Bipyridine-kappa2N,N')(dichromato-kappaO)copper(II). AB - The title compound, [Cu(Cr(2)O(7))(C(10)H(8)N(2))(2)], a new mixed-metal molecular compound, contains isolated molecular units, each comprised of one Cu(II) atom coordinated to two 2,2'-bipyridine ligands and also to an oxygen vertex of a dichromate anion. The Cu(II) atom has an approximate trigonal bipyramidal geometry, which is consistent with previous studies. Both enantiomers of the chiral complex molecule are present and are related by inversion centers. In a reported pyridine analogue, achiral [Cu(Cr(2)O(7))(pyridine)(4)] chains pack in the non-centrosymmetric space group Pna2(1). Differences in the organic ligands influence the chirality and dimensionality of the Cu-Cr(2)O(7) bonding. PMID- 11932518 TI - Bis(isoquinoline-1-carboxylato-kappa2O,N)(1,10-phenanthroline-kappa2N,N')zinc(II) dimethylformamide sesquisolvate. AB - The asymmetric unit of the title compound, [Zn(C(10)H(6)NO(2))(2)(C(12)H(8)N(2))] x 1.5C(3)H(7)NO, contains one monomeric zinc complex and 1.5 disordered dimethylformamide solvate molecules. The Zn atom is coordinated to one 1,10 phenanthroline ligand and to two isoquinolinecarboxylate anions (IQC-) via their N and O atoms. The complex exhibits a distorted octahedral geometry around the Zn(II) atom, with the apical positions occupied by the O atoms of the IQC- ligands. The Zn atom lies 0.049 (1) A out of the basal plane. The crystal packing is characterized by several hydrogen bonds. PMID- 11932519 TI - A cyclic tetramer of [Ag(1,4,7-trithiacyclononane)]+, cyclo-tetrakis(mu-1,4,7 trithiacyclononane-kappa3S1,S4,S7:kappaS1)tetrasilver(I) tetrakis(trifluoromethanesulfonate) nitromethane disolvate. AB - (1,4,7-Trithiacyclononane)silver trifluoromethanesulfonate crystallizes in a tetrameric form from nitromethane, to give the title compound, [Ag(4)(C(6)H(12)S(3))(4)](CF(3)SO(3))(4) x 2CH(3)NO(2). The complex cation consists of four [AgL]+ units (L is 1,4,7-trithiacyclononane), with four Ag-S-Ag bridges forming a cyclic tetramer. The almost planar Ag(4)S(4) ring takes an octagonal form. PMID- 11932520 TI - A lithium complex of pyridine-2,6-dicarboxylic acid. AB - The crystal structure of catena-poly[[(6-carboxypyridine-2-carboxylato kappa3O,N,O')lithium(I)]-mu-aqua-kappa2O:O], [Li(C(7)H(4)NO(4))(H(2)O)](n), contains the Li(+) ion coordinated to two O atoms and the N atom of the 6 carboxypyridine-2-carboxylate ligand, and to two water O atoms, forming a pentavalent coordination geometry. The molecule resides on a mirror plane which contains the Li and N atoms, the para-CH unit, and the O atom of the coordinated water molecule. The O atom of the water molecule is coordinated to two Li atoms, forming an infinite polymeric chain. PMID- 11932521 TI - Tin-halide compounds. IV1. tert-butyltrichlorotin (IV). AB - The molecules of the title compound, [Sn(C(4)H(9))Cl(3)], adopt an eclipsed conformation with respect to the C and Cl atoms, and possess crystallographic C(s) symmetry. The molecular structure and geometric parameters are comparable with those of related organotin trihalides. PMID- 11932522 TI - Bis(tetramethylammonium) hexaaquadodeca-mu-bromo-octahedro-hexatantalum tetrabromide dihydrate. AB - The novel title compound, [(CH(3))(4)N](2)[Ta(6)Br(12)(H(2)O)(6)]Br(4) x 2H(2)O, with a [Ta(6)Br(12)]2+ cluster unit, has been prepared and structurally characterized. The compound crystallizes in space group C2/c, with a twofold axis passing through the cluster and the centre of symmetry located between the clusters. The nearest neighbouring cluster units are aligned along the crystallographic c axis, forming a one-dimensional chain pattern. PMID- 11932523 TI - Two nickel complexes stabilized by nitrate counter-ions. AB - Two new nickel nitrates, diaquabis(3,4,7,8-tetramethyl-1,10-phenanthroline kappa2N,N')nickel(II) dinitrate methanol solvate, [Ni(C(16)H(16)N(2))(2)(H(2)O)(2)](NO(3))(2) x CH(4)O, (I), and triaqua[2,4,6 tris(2-pyridyl)-1,3,5-triazine-kappa3N1,N2,N6]nickel(II) dinitrate trihydrate, [Ni(C(18)H(12)N(6))(H(2)O)(3)](NO(3))(2) x 3H(2)O, (II), are reported. In both structures, the cation is octahedrally coordinated, to two bidentate 3,4,7,8 tetramethyl-1,10-phenanthroline (tmp) and two water molecules in (I), and to one tridentate 2,4,6-tris(2-pyridyl)-1,3,5-triazine (tpt) and three water molecules in (II). Both structures are stabilized by extensive hydrogen-bonding interactions. PMID- 11932525 TI - Bis(3,5-dimethylpyrazole-kappaN2)silver(I) nitrate. AB - The two independent bis(3,5-dimethylpyrazole)silver(I) cations in crystalline [Ag(C(5)H(7)N(2))(2)]NO(3) display N-Ag-N angles of 175.51 (14) and 174.44 (13) degrees, and an average Ag-N distance of 2.124 (5) A. The nitrate anion is situated between [Ag(C(5)H(7)N(2))(2)]+ units and interacts via hydrogen bonds with the NH groups. The two 3,5-dimethylpyrazole ligands are trans about the silver center. Only a small deviation from linearity is observed in the coordination around silver. PMID- 11932524 TI - Bis[hydrogen 1,1'-bis(diphenylphosphinato)ferrocenium] octachlorodiantimony(III). AB - In the title compound, [Fe(C(34)H(29)O(2)P(2))](2)[Sb(2)Cl(8)], the discrete centrosymmetric [Sb(2)Cl(8)]2- anions are formed from two edge-shared square pyramids of Cl atoms about each Sb atom. Within the cation, the two diphenylphosphinate groups share one H atom and the ferrocene cyclopentadienyl rings are in a staggered conformation, with the average value of the twist angle being 46 degrees. In the crystal, each [Sb(2)Cl(8)]2- anion is involved in eight C-H* * *Cl interactions with four surrounding cations and these interactions interconnect the ions to form molecular columns along the a direction. PMID- 11932526 TI - The seven-membered nickelacycle [NiBr=(o-CH=C(CF3)C6H4CH2PPh2 kappa2C,P)(PPh(CH2Ph)2)]. AB - The crystal and molecular structures of the title compound, 3-bromo-3 (dibenzylphenylphosphonio)-2,2-diphenyl-5-trifluoromethyl-1H benzo[e][1,2]phosphanickelepine, [NiBr(C(22)H(17)F(3)P)(C(20)H(19)P)], which was obtained as the major regioisomer from insertion of HCCCF(3) into the Ni-C bond of the five-membered phosphanickelacycle [NiBr(o-C(6)H(4)CH(2)PPh(2) kappa2C,P)(PPh(CH(2)Ph)(2))], have been determined. Principal geometric data include the Ni-X bond lengths Ni-Br 2.3343 (4) A, Ni-P 2.1867 (7) and 2.2094 (7) A, and Ni-C 1.882 (3) A, and the two trans angles P-Ni-P 171.55 (3) degrees and Br-Ni-C 176.88 (9) degrees. PMID- 11932527 TI - Tetra-mu-acetato-kappa2O:O'-bis[(4-phenylpyridine-kappaN)copper(II)]. AB - The title compound, [Cu(2)(C(2)H(3)O(2))(4)(C(11)H(9)N)(2)] or [Cu(2)(MeCO(2))(4)(phpy)(2)] (phpy is 4-phenylpyridine), consists of centrosymmetric dimers in which the CuII atoms display a square-pyramidal CuO(4)N coordination, with four acetate O atoms in the basal plane [Cu-O 1.975 (3)-1.987 (3) A] and the phpy N atom in the apical position [Cu-N 2.150 (3) A]. The Cu atoms are 2.654 (1) A apart and are bridged by four acetate groups. The discrete dimers are extended into a three-dimensional supramolecular array through intermolecular pi-pi-stacking interactions. PMID- 11932528 TI - A complex containing three different kinds of Ru-N bonds: ethoxydinitronitrosyl(N,N,N',N'-tetramethylethylenediamine kappa2N,N')ruthenium(II). AB - The octahedral title compound, [Ru(C(2)H(5)O)(NO)(NO(2))(2)(C(6)H(16)N(2))], crystallizes in the rhombohedral space group P3(1) with an ethoxy ligand axially coordinated trans to the nitrosyl ligand. The RuII ion is equatorially coordinated by a tetramethylethylenediamine group acting as a bidentate ligand, and to two nitro moieties whose planes are tilted with respect to the mean equatorial plane. Each nitrogen ligand bonded to the metallic centre has a different hybridization state. PMID- 11932529 TI - cis-[1,2-Bis(diisopropylphosphino-kappaP)-1,2-dicarba-closo dodecaborane]dichloroplatinum(II) dichloromethane hemisolvate. AB - The asymmetric unit of the title complex, [PtCl(2)(C(14)H(38)B(10)P(2))] x 0.5CH(2)Cl(2) or cis-[PtCl(2)(1,2-(P(i)Pr(2))(2)-1,2-C(2)B(10)H(10))] x 0.5CH(2)Cl(2), contains one disordered solvent molecule and two molecules of the complex, in which each PtII atom displays slightly distorted square-planar coordination geometry. The P atoms connected to the cage C atoms are coordinated to the PtII atom. The Pt-P distances vary slightly [2.215 (3) and 2.235 (4) A] and the Pt-Cl distances are equal [2.348 (3) and 2.353 (5) A]. PMID- 11932530 TI - (BETS)2[Fe(tdas)2]2: a new metal in the molecular conductor family. AB - The structure of bis[4,5-ethylenedithio-2-(4,5-ethylenedithio-1,3 diselenacyclopent-4-en-2-ylidene)-1,3-diselenacyclopent-4-enium] bis(mu-1,2,5 thiadiazole-3,4-dithiolato-kappa3S4,S5:S4)bis[(1,2,5-thiadiazole-3,4-dithiolato kappa2S4,S5)iron(III)], (BETS)(2)[Fe(tdas)(2)](2) [BETS is alternatively called bis(ethylenedithio)tetraselenafulvalenium] or (C(10)H(8)S(4)Se(4))(2)[(Fe(C(2)N(2)S(3))(2))(2)], consists of segregated columns of dimers of BETS and columns of dimers of [Fe(tdas)(2)]. Each dimer displays inversion symmetry. Numerous chalcogen-chalcogen contacts are observed within and between the columns, producing a network of interactions responsible for the metal-like behaviour of the compound. PMID- 11932531 TI - Tetrakis[1-(3-chloropropyl)-1,2,4-triazole-kappaN4]bis(tetrafluoroborato kappaF)copper(II). AB - In the title complex, [Cu(BF(4))(2)(1tpc)(4)] [1tpc is 1-(3-chloropropyl)-1,2,4 triazole, C(5)H(8)ClN(3)], the copper(II) centres reside in a tetragonally distorted octahedral coordination environment. Four 1tpc ligands are coordinated to the metal atom via the N4 atom of the triazole rings in a square-planar arrangement, with Cu-N bond lengths in the range 2.002 (2)-2.019 (2) A. Two tetrafluoroborate anions, in the axial positions above and below the square plane, are weakly coordinated to the copper(II) centre, with Cu-F distances of 2.4009 (18) and 2.5096 (18) A. PMID- 11932532 TI - Low-dimensional compounds containing cyano groups. I. catena Poly[dicyanoargentato(I)-kappaN]bis(4-methylpyridine-kappaN)copper(II)]-mu dicyanoargentato(I)-kappa2N:N']. AB - The structure of the title compound, [Cu(C(6)H(7)N)(2)(Ag(CN)(2)(2)](n), is made up of neutral zigzag chains of [-NC-Ag-CN-Cu(4-Mepy)(2)(Ag(CN)(2))-NC-Ag-CN-] (4 Mepy is 4-methylpyridine). Neighbouring chains are linked by weak argentophilic interactions, with Ag.Ag distances of 3.2322 (12) A. The Cu atom, which lies on a twofold rotation axis, is pentacoordinated by one monodentate Ag(CN)(2)(-) anion [Cu-N 1.985 (3) A], the atoms of which lie on the same rotation axis, and by bridging dicyanoargentate anions [2 x Cu-N 2.0827 (19) A], with Ag atoms on inversion centres. The coordination polyhedron is completed by two 4-Mepy molecules [2 x Cu-N 2.038 (2) A], which occupy the axial positions of a distorted trigonal bipyramid. PMID- 11932533 TI - Undecacarbonyl(methylcyclopentadienyl)-tetrahedro-triiridiummolybdenum, undecacarbonyl(tetramethylcyclopentadienyl)-tetrahedro-triiridiummolybdenum and undecacarbonyl(pentamethylcyclopentadienyl)-tetrahedro-triiridiummolybdenum. AB - The three title compounds tri-mu-carbonyl-1:2kappa2C;1:3kappa2C;2:3kappa2C octacarbonyl-1kappaC,2kappa2C,3kappa2C,4kappa3C-eta5-methylcyclopentadienyl tetrahedro-triiridiummolybdenum(3 Ir-Ir)(3 Ir-Mo), tri-mu-carbonyl 1:2kappa2C;1:3kappa2C;2:3kappa2C-octacarbonyl-1kappaC,2kappa2C,3kappa2C,4kappa3C eta5-tetramethylcyclopentadienyl-tetrahedro-triiridiummolybdenum(3 Ir-Ir)(3 Ir Mo) and tri-mu-carbonyl-1:2kappa2C;1:3kappa2C;2:3kappa2C-octacarbonyl 1kappaC,2kappa2C,3kappa2C,4kappa3C-eta5-pentamethylcyclopentadienyl-tetrahedro triiridiummolybdenum(3 Ir-Ir)(3 Ir-Mo), [MoIr(3)(eta5-C(5)H(5-n)Me(n))(mu CO)(3)(CO)(8)], where n = 1, 4 or 5, have a pseudotetrahedral MoIr(3) core geometry, with a eta5-C(5)H(5-n)Me(n) group ligating the Mo atom, bridging carbonyls spanning the edges of an MoIr(2) face, and eight terminally bound carbonyls. PMID- 11932534 TI - Two thiourea-containing gold(I) complexes. AB - The crystal structures of two salts of bis(thiourea)gold(I) complexes, namely bis(thiourea-kappaS)gold(I) chloride, [Au(CH(4)N(2)S)(2)]Cl, (I), and bis[bis(thiourea-kappaS)gold(I)] sulfate, [Au(CH(4)N(2)S)(2)](2)SO(4), (II), have been determined. The chloride salt, (I), is isomorphous with the corresponding bromide salt, although there are differences in the bonding. The Au(I) ion is located on an inversion centre and coordinated by two symmetry-related thiourea ligands through the lone pairs on their S atoms [Au-S 2.278 (2) A and Au-S-C 105.3 (2) degrees ]. The sulfate salt, (II), crystallizes with four independent [Au(CH(4)N(2)S)(2)]+ cations per asymmetric unit, all with nearly linear S-Au-S bonding. The cations in (II) have similar conformations to that found for (I). The Au-S distances range from 2.276 (3) to 2.287 (3) A and the Au-S-C angles from 173.5 (1) to 177.7 (1) degrees. These data are relevant in interpreting different electrochemical processes where gold-thiourea species are formed. PMID- 11932535 TI - [P,P-Di-tert-butyl-N-trimethylsilyl-P-(trimethylsilylamino)phosphine imidato kappa2N,N']bis(pyridine-kappaN)lithium(I). AB - In the title compound, [Li(C(14)H(36)N(2)PSi(2))(C(5)H(5)N)(2)], the bulky chelating monoanionic P,P-di-tert-butyl-N-trimethylsilyl-P (trimethylsilylamino)phosphine imidate ligand and two pyridine ligands bind to Li in a pseudo-tetrahedral arrangement with twofold symmetry. The Li-N(phosphine) distance is 2.048 (5) A, while the Li.P distance is 2.520 (6) A. PMID- 11932536 TI - Aquachloro[N,N'-ethylenebis(salicylideneiminato)]manganese(III). AB - The title compound, aquachloro(2,2'-[1,2 ethanediylbis(nitrilomethylidyne)]diphenolato kappa4O,N,N',O')manganese(III),[MnCl(C(16)H(14)N(2)O(2))(H(2)O)], is a neutral manganese(III) complex with a pseudo-octahedral metal centre. The equatorial plane comprises the four donor atoms of the tetradentate Schiff base ligand [Mn-O 1.886 (4) and 1.893 (4) A, and Mn-N 1.978 (5) and 1.982 (5) A], with a water molecule [Mn-O 2.383 (4) A] and a Cl- ligand [Mn-Cl 2.4680 (16) A] completing the coordination sphere. The distorted geometry is highlighted by the marked displacement of the Mn(III) ion out of the least-squares plane of the four Schiff base donor atoms by 0.165 (2) A. These monomeric Mn(III) centres are then linked into a polymeric array via hydrogen bonds between the coordinated water molecule and the phenolic O-atom donors of an adjacent Mn(III) centre [O-H.O 2.789 (5) and 2.881 (5) A]. PMID- 11932538 TI - CsMoO2(HO3P-CH2-PO3): a new metallodiphosphonate with a hybrid framework. AB - Crystals of caesium molybdenomethylenediphosphonate, [CsMoO(2)(CH(3)O(6)P(2))], were hydrothermally synthesized at 473 K. The monoclinic structure, as determined from single-crystal X-ray diffraction, is two-dimensional and consists of stacked mixed layers of corner-sharing tetrahedral diphosphonate groups and MoO(6) octahedra, between which Cs+ cations are intercalated. PMID- 11932539 TI - [5-Bromo-N-(2-carboxylatophenyl)salicylideniminato]dimethyltin(IV). AB - The structure of the title dimethyltin(IV) complex, [2-(5-bromo-2 oxidobenzylideneamino)benzoato-kappa3O,N,O']dimethyltin(IV), [Sn(CH(3))(2)(C(14)H(8)BrNO(3))], features centrosymmetric dimers disposed about a central Sn(2)O(2) core. Each Sn centre has seven-coordinate pentagonal bipyramidal geometry, taking into account two moderately long Sn-O contacts about an inversion centre [2.679 (4) and 2.981 (4) A]. The methyl groups are in an axial orientation. PMID- 11932537 TI - Layered (ethylenediamine-kappa2N,N')cobalt(II) molybdate(VI). AB - The structure of poly[[(ethylenediamine-kappa2N,N')cobalt(II)]-mu tetraoxomolybdato(VI)], [Co(C(2)H(8)N(2))MoO(4)](n) or [CoMoO(4)(C(2)H(8)N(2))](n), is composed of puckered layers constructed from MoO(4) tetrahedra and CoN(2)O(4) octahedra, with the ethylenediamine ligand coordinated to the Co atom in a cis fashion. Each pair of cobalt sites forms a binuclear edge-sharing unit through a (Co(2)O(2)) interaction. The binuclear octahedral units are interconnected through the bridging MoO(4) tetrahedra into a layer structure. PMID- 11932540 TI - Triaqua(2,2'-bipyridine-kappa2N,N')(nitrato-kappaO)manganese(II) nitrate. AB - The crystal structure of the title compound, [Mn(NO(3))(C(10)H(8)N(2))(H(2)O)(3)]NO(3), contains a monomeric [Mn(NO(3))(bpy)(H(2)O)(3)]+ cation (bpy is 2,2'-bipyridine) and a nitrate anion. The Mn(II) ion is coordinated by one chelating bpy [Mn-N 2.241 (3) and 2.259 (3) A], three water molecules [Mn-O 2.120 (3)-2.188 (3) A] and a nitrate ligand [Mn-O 2.228 (2) A] in a distorted octahedral geometry. There are O.H-O hydrogen-bonding interactions between the ligated water molecules and the ligated and unligated nitrate anions, resulting in double columns of stacked cations and anions. PMID- 11932541 TI - catena-Poly[diaquabis(5-chloropyridinium-2-olato-kappaO)copper(II)]-mu-pyrazine kappa2N:N'] diperchlorate]. AB - In the title compound, [Cu(C(5)H(4)ClNO)(2)(C(4)H(4)N(2))(H(2)O)(2)](ClO(4))(2), the Cu atom, which lies on an inversion centre, has an octahedral environment. The pyrazine ligand also lies about an inversion centre and links adjacent Cu atoms into a chain running along the b axis; perchlorate anions occupy the space between the chains, and the chains use the coordinated water molecules to link to the anions, resulting in a hydrogen-bonded ribbon structure. The donor 5-chloro-2 hydroxypyridine ligand exists in the zwitterionic form, i.e. 5-chloropyridinium-2 olate. PMID- 11932542 TI - Hydrogen-bonded molecular ladders in S-(4-nitrophenyl)thioglycolic acid. AB - Molecules of the title compound, [(4-nitrophenyl)sulfanyl]acetic acid, C(8)H(7)NO(4)S, are linked by paired O-H* * *O hydrogen bonds [H.O 1.81 A, O.O 2.6456 (15) A and O-H* * *O 178 degrees ] into centrosymmetric dimers containing an R(2)2(8) motif. A single C-H* * *O hydrogen bond having a nitro O atom as acceptor [H* * *O 2.47 A, 3.3018 (19) A and C-H.O 147 degrees ] links the dimers into a molecular ladder, and neighbouring ladders are weakly linked into sheets by aromatic pi-pi-stacking interactions. PMID- 11932543 TI - Adducts of 1,1,1-tris(4-hydroxyphenyl)ethane with diamines: three-dimensional hydrogen-bonded frameworks formed with 1,6-diaminohexane and 2,2'-bipyridyl. AB - The adduct 1,6-diaminohexane-1,1,1-tris(4-hydroxyphenyl)ethane (1/2) is a salt (hexane-1,6-diyldiammonium-4-[1,1-bis(4-hydroxyphenyl)ethyl]phenolate (1/2)), C(6)H(18)N(2)2+ x 2C(20)H(17)O(3)(-), in which the cation lies across a centre of inversion in space group P-1. The anions are linked by two short O-H* * *O hydrogen bonds [H* * *O 1.74 and 1.76 A, O.O 2.5702 (12) and 2.5855 (12) A, and O H* * *O 168 and 169 degrees ] into a chain containing two types of R(2)2(24) ring. Each cation is linked to four different anion chains by three N-H* * *O hydrogen bonds [H* * *O 1.76-2.06 A, N* * *O 2.6749 (14)-2.9159 (14) A and N-H* * *O 156-172 degrees ]. In the adduct 2,2'-bipyridyl-1,1,1-tris(4 hydroxyphenyl)ethane (1/2), C(10)H(8)N(2) x 2C(20)H(18)O(3), the neutral diamine lies across a centre of inversion in space group P2(1)/n. The tris(phenol) molecules are linked by two O-H* * *O hydrogen bonds [H* * *O both 1.90 A, O* * *O 2.7303 (14) and 2.7415 (15) A, and O-H* * *O 173 and 176 degrees ] into sheets built from R(4)4(38) rings. Pairs of tris(phenol) sheets are linked via the diamine by means of a single O-H* * *N hydrogen bond [H* * *N 1.97 A, O* * *N 2.7833 (16) A and O-H* * *N 163 degrees ]. PMID- 11932544 TI - A quasi-diamondoid hydrogen-bonded framework in anhydrous sulfanilic acid. AB - The title compound (C(6)H(7)NO(3)S) exists as a zwitterion (4 ammoniobenzenesulfonate), +H(3)NC(6)H(4)SO(3)-, and these units are linked into a three-dimensional framework by two distinct two-centre N-H* * *O hydrogen bonds [H* * *O 1.84 and 1.87 A; N* * *O 2.767 (2) and 2.746 (2) A; N-H* * *O 166 and 172 degrees ] and a planar three-centre N-H* * *(O)(2) hydrogen bond [H* * *O 2.03 and 2.37 A; N* * *O 2.816 (2) and 2.877 (2) A; N-H* * *O 162 and 111 degrees; O* * *H* * *O 86 degrees ]. PMID- 11932545 TI - Design of peptides with alpha,beta-dehydro residues: pseudo-tripeptide N benzyloxycarbonyl-DeltaLeu-L-Ala-L-Leu-OCH3. AB - The title peptide N-benzyloxycarbonyl-DeltaLeu-L-Ala-L-Leu-OCH(3) [methyl N (benzyloxycarbonyl)-alpha,beta-dehydroleucyl-L-alanyl-L-leucinate], C(24)H(35)N(3)O(6), was synthesized in the solution phase. The peptide adopts a type II' beta-turn conformation which is stabilized by an intramolecular 4 --> 1 N-H* * *O hydrogen bond. The crystal packing is stabilized by two intermolecular N-H* * *O hydrogen bonds. PMID- 11932546 TI - Reactive intermediates in peptide synthesis. ortho-Nitrophenyl Nalpha-para toluenesulfonyl-alpha-aminoisobutyrate. AB - The preparation, characterization, and molecular and crystal structures of the title compound [IUPAC name: 2-nitrophenyl 2-methyl-2-(para toluenesulfonylamino)propanoate], C(17)H(18)N(2)O(6)S, are reported. The phenyl group is almost perpendicular to the plane of the adjacent ester moiety. One O atom of the nitro group is wedged between the two ester O atoms. The implications of this peculiar conformation for the chemistry of ortho-nitrophenyl esters in peptide synthesis are discussed. PMID- 11932547 TI - 3,4-(4-Methoxybenzo):8,9-benzobicyclo[4.4.1]undeca-3,8-dien-11-one ethylene acetal. AB - The title compound, 5-methoxyspiro[tetracyclo[8.8.1.0(3,8).0(12,17)]nonadeca 3,5,7,12,14,16-hexene-19,2'-[1,3]dioxolane], C(22)H(24)O(3), exhibits a twin chair conformation with the aromatic rings overlying each other. Comparison of the dihedral angle between these two rings with those from previously reported [3.3]orthocyclophanes of this type suggests the presence of a weak attractive charge-transfer interaction between the two, different, stacked arenes. PMID- 11932548 TI - 1-Methyl-5,6-diphenylpyrazine-2(1H)-thione. AB - The title compound, C(17)H(14)N(2)S, crystallizes in a triclinic unit cell, with two crystallographically independent molecules in the asymmetric unit. The two independent molecules pack in the same sense and form segregated layers along the c axis. The crystal is light-stable and no dimers are formed under irradiation. The intermolecular distances between the potential reactive centers (the C-3 and C-5 ring positions) are 4.093 (4) and 5.643 (4) A for molecule A, and 4.081 (4) and 5.614 (4) A for molecule B. PMID- 11932549 TI - Two triethylammonium ion complexes of monoanionic calix[4]arene. AB - Two complexes between mono-deprotonated calix[4]arene and Et(3)HN+ are reported. The first, triethylammonium 26,27,28 trihydroxypentacyclo[19.3.1.1(3,7).1(9,13).1(15,19)]octacosa 1(25),3,5,7(28),9,11,13(27),15,17,19(26),21,23-dodecaen-25-olate, C(6)H(16)N+ x C(28)H(23)O(4)-, comprises only the cationic and anionic species, whereas the second, tris(triethylammonium) tris[26,27,28 trihydroxypentacyclo[19.3.1.1(3,7).1(9,13).1(15,19)]octacosa 1(25),3,5,7(28),9,11,13(27),15,17,19(26),21,23-dodecaen-25-olate] acetonitrile solvate, 3C(6)H(16)N+ x 3C(28)H(23)O(4)(-) x C(2)H(3)N, comprises one acetonitrile solvent molecule for three such units. In both cases, the units are stacked in columns so that the ammonium cation is hydrogen bonded to a phenolic or phenolate O atom of one molecule and included in the hydrophobic cavity of another neighbouring calixarene. The short contacts present indicate that cation* * *pi and C-H* * *pi interactions are likely to be involved in the inclusion phenomena. PMID- 11932550 TI - cis-Dioxocyclam. AB - Molecules of 1,4,8,11-tetraazacyclotetradecane-5,7-dione, or cis-dioxocyclam, C(10)H(20)N(4)O(2), lie across mirror planes in space group Cmca; the crystal structure reveals interleaved columns of cis-dioxocyclam molecules along the 2(1) screw axis parallel to the crystallographic b axis. The columns are interconnected in a chain-like arrangement by an amido hydrogen-bonding network (N* * *O = 2.816 A) and an amino hydrogen-bonding network (N* * *N = 3.193 A). The intracolumn spacing is 9.02 A. PMID- 11932551 TI - The supramolecular structure of pyridine-2,6-dicarboxylic acid. AB - The structure of pyridine-2,6-dicarboxylic acid, C(7)H(5)NO(4), has been determined at 0.71 A resolution. The molecule is located on a site with mirror symmetry. A one-dimensional supramolecular structure is stabilized in the solid state through a strong symmetric double hydrogen bond, with H* * *O distances of 1.86 (3) A and O-H* * *O angles of 167 (3) and 171 (5) degrees. This arrangement is similar but not identical to that reported for the isoelectronic isophthalic acid (benzene-1,3-dicarboxylic acid). PMID- 11932552 TI - The dimer, trimer and 1,2,4-trithiolane of adamantanethione. AB - The molecules of dispiro[1,3-dithietane-2,2':4,2"-diadamantane], C(20)H(28)S(2), have crystallographic C(i) symmetry, as well as local D(2h) symmetry, and a planar 1,3-dithietane ring. The molecules of trispiro[1,3,5-trithiane 2,2':4,2":6,2"'-triadamantane], C(30)H(42)S(3), have approximate C(2) symmetry and the 1,3,5-trithiane ring has a twist-boat conformation. The C-S-C bond angles within the ring are about 8 degrees larger than observed in most related 1,3,5 trithiane structures. In dispiro[1,2,4-trithiolane-3,2':5,2"-diadamantane], C(20)H(28)S(3), the molecules have local C(2) symmetry and the 1,2,4-trithiolane ring has a half-chair conformation. PMID- 11932553 TI - The crystal packing of 4,7-dichloro- and 4,7-dibromobenzo[c]furazan 1-oxide. AB - The molecular structures of 4,7-dichlorobenzo[c]furazan 1-oxide, C(6)H(2)Cl(2)N(2)O(2), (I), and 4,7-dibromobenzo[c]furazan 1-oxide, C(6)H(2)Br(2)N(2)O(2), (II), are normal. Compound (I) occurs in two polymorphic forms. One polymorph contains one molecule in the asymmetric unit, organized into two-dimensional sheets involving intermolecular N* * *Cl and O* * *Cl interactions. The second polymorph has three molecules in the asymmetric unit, organized into two crystallographically different two-dimensional sheets with similar interactions. Compound (II) is isomorphous with the second polymorph of (I). The three two-dimensional sheets in the two polymorphs comprise a set of three two-dimensional polymorphic arrangements. PMID- 11932554 TI - 1-Acetyl-3-[1-(4-methoxyphenyl)-2-oxopropylidene]-1H-indol-2(3H)-one: a twinned crystal. AB - The crystal of the title compound, C(20)H(17)NO(4), which was used for collecting intensity data was twinned. Each of the two crystallographically independent molecules in the asymmetric unit has a planar indole moiety perpendicular to a planar oxopropyl moiety. The distribution of the bonds at the central C atom joining the oxopropyl, phenyl and indole substituents is also planar. The packing is stabilized by intermolecular C-H* * *O interactions, as well as by dipole dipole and van der Waals interactions. PMID- 11932555 TI - Two-component molecular crystals composed of chloronitrobenzoic acids and 4 aminopyridine. AB - The crystal structures of the four isomeric organic salts 4-aminopyridinium 2 chloro-4-nitrobenzoate, (I), 4-aminopyridinium 2-chloro-5-nitrobenzoate, (II), 4 aminopyridinium 5-chloro-2-nitrobenzoate, (III), and 4-aminopyridinium 4-chloro-2 nitrobenzoate, (IV), all C(5)H(7)N(2)+ x C(7)H(3)ClNO(4)-, are presented. Compound (I) has one intramolecular hydrogen bond, one intermolecular C-H* * *O hydrogen bond and pi-pi-stacking interactions. Compound (II) has N-H* * *O, C-H* * *O and C-H* * *Cl hydrogen bonds, and Cl* * *O-C electrostatic interactions. Compound (III) has N-H* * *O and C-H* * *O hydrogen bonds. Compound (IV) has a pi pi-stacking interaction, but no C-H* * *O hydrogen bonds. PMID- 11932556 TI - Triorganophosphine-dithiomonometaphosphoryl halides. AB - The title compounds, ethyldiphenylphosphine-dithiomonometaphosphoryl chloride, EtPh(2)P-->PS(2)Cl, C(14)H(15)ClP(2)S(2), (I), and tris-n-propylphosphine dithiomonometaphosphoryl chloride and bromide, nPr(3)P-->PS(2)Cl, C(9)H(21)ClP(2)S(2), (II), and nPr(3)P-->PS(2)Br, C(9)H(21)BrP(2)S(2), (III), respectively, are the first phosphine-stabilized dithiomonometaphosphoryl halides to be structurally characterized. In the tris-n-propylphosphine derivatives, the central P-->P donor-acceptor bond becomes longer in the order bromo < chloro < fluoro. Substitution of the tris-n-propylphosphine group in (II) by the more bulky ethyldiphenylphosphine group also leads to a longer P-->P bond. These structural features agree with the observed 31P NMR data. In (II) and (III), the central P-P bond coincides with the crystallographic threefold axis, entailing site-occupational disorder for the S(2)Y group. PMID- 11932557 TI - Ghostbursting: a novel neuronal burst mechanism. AB - Pyramidal cells in the electrosensory lateral line lobe (ELL) of weakly electric fish have been observed to produce high-frequency burst discharge with constant depolarizing current (Turner et al., 1994). We present a two-compartment model of an ELL pyramidal cell that produces burst discharges similar to those seen in experiments. The burst mechanism involves a slowly changing interaction between the somatic and dendritic action potentials. Burst termination occurs when the trajectory of the system is reinjected in phase space near the "ghost" of a saddle-node bifurcation of fixed points. The burst trajectory reinjection is studied using quasi-static bifurcation theory, that shows a period doubling transition in the fast subsystem as the cause of burst termination. As the applied depolarization is increased, the model exhibits first resting, then tonic firing, and finally chaotic bursting behavior, in contrast with many other burst models. The transition between tonic firing and burst firing is due to a saddle node bifurcation of limit cycles. Analysis of this bifurcation shows that the route to chaos in these neurons is type I intermittency, and we present experimental analysis of ELL pyramidal cell burst trains that support this model prediction. By varying parameters in a way that changes the positions of both saddle-node bifurcations in parameter space, we produce a wide gallery of burst patterns, which span a significant range of burst time scales. PMID- 11932558 TI - An estimator for the electrotonic size of neurons independent of charge equalization time constants. AB - Electrotonic properties are important aspects of neuronal function but have been difficult to estimate without accurate morphological reconstruction. The complexity of the branching dendritic cables often gives charging curves composed of a very large number of exponential functions, making it difficult to distinguish the time constants that are needed for electrotonic estimates. We describe an estimator P for the electrotonic size of neurons based on simple measures from voltage and current clamp recordings that does not rely on the higher rank exponential components of the response. Our estimator gives a bounded scale for the electrotonic size of the cell and can be used for categorization and comparison when morphology is not available. PMID- 11932559 TI - A spiking neuron model for binocular rivalry. AB - We present a biologically plausible model of binocular rivalry consisting of a network of Hodgkin-Huxley type neurons. Our model accounts for the experimentally and psychophysically observed phenomena: (1) it reproduces the distribution of dominance durations seen in both humans and primates, (2) it exhibits a lack of correlation between lengths of successive dominance durations, (3) variation of stimulus strength to one eye influences only the mean dominance duration of the contralateral eye, not the mean dominance duration of the ipsilateral eye, (4) increasing both stimuli strengths in parallel decreases the mean dominance durations. We have also derived a reduced population rate model from our spiking model from which explicit expressions for the dependence of the dominance durations on input strengths are analytically calculated. We also use this reduced model to derive an expression for the distribution of dominance durations seen within an individual. PMID- 11932561 TI - Effect of growth hormone on exercise tolerance in children with cystic fibrosis. AB - PURPOSE: The effect of growth hormone (GH) treatment on exercise tolerance in children with cystic fibrosis was investigated. METHODS: 10 prepubertal children (mean +/- SD; age: 12.1 +/- 1.7 yr; height: 137.4 +/- 9.2 cm; body mass: 27.8 +/- 4.2 kg; forced expiratory volume in 1 s (FEV1): 68 +/- 22% predicted) were randomly assigned to either control period (CON, standard therapy) or recombinant human growth hormone (GH) period (additional GH treatment, 0.11-0.14 IU.kg-1, daily, s.c.) for the first 6 months, and then assigned to the other period for the next 6 months. At study entry and after each period, anthropometric data, pulmonary function, and exercise capacity (peak exercise capacity, .VO(2peak), and isokinetic muscle strength) were measured. RESULTS: Changes in height (+4.3 +/- 1.0 cm), total body mass (+2.2 +/- 0.8 kg), and lean body mass (LBM, +2.9 +/- 0.7 kg) were significantly higher (P < 0.01) after GH treatment compared with CON. Pulmonary function did not significantly change in either of the periods. In contrast to CON, GH treatment improved absolute .VO(2peak) (+19%, P < 0.01), peak ventilation (+14%, P < 0.01), and peak oxygen pulse (+18%, P < 0.01). Analysis of variance revealed that most of the changes (71%) in .VO(2peak) could be explained by those in LBM and FEV1 (P = 0.001). CONCLUSION: GH treatment clearly improved exercise tolerance, presumably resulting from the combined effects of GH on the muscular, cardiovascular, and pulmonary capacity. PMID- 11932562 TI - Management of Achilles tendinopathy by ultrasound-guided percutaneous tenotomy. AB - PURPOSE: To report the middle to long-term results of ultrasound-guided percutaneous longitudinal tenotomy of the Achilles tendon METHOD: Seventy-five athletes with unilateral Achilles tendinopathy underwent ultrasound-guided percutaneous longitudinal tenotomy under local anesthetic infiltration after failure of conservative management. Sixty-three patients were reviewed at least 36 months after the operation (51 +/- 18.2 months). RESULTS: Thirty-five patients were rated excellent, 12 good, 9 fair, and 7 poor. Nine of the 16 patients with a fair or poor result underwent a formal exploration of the Achilles tendon 7-12 months after the index procedure. The operated tendons remained thickened and the ultrasonographic appearance of operated tendons remained abnormal even 8 yr after the operation, without interfering with physical training. Isometric maximal muscle strength and isometric endurance gradually returned to values similar to their contralateral unoperated tendon. CONCLUSIONS: Percutaneous longitudinal ultrasound-guided internal tenotomy is simple, can be performed on an outpatient basis, requires minimal follow-up care, does not hinder further surgery should it be unsuccessful, and, in our experience, has produced no significant complications. It should be considered in the management of chronic Achilles tendinopathy after failure of conservative management. However, patients should be advised that, if they suffer from diffuse or multinodular tendinopathy or from pantendinopathy, a formal surgical exploration with stripping of the paratenon and multiple longitudinal tenotomies may be preferable. PMID- 11932560 TI - Summation of spatiotemporal input patterns in leaky integrate-and-fire neurons: application to neurons in the cochlear nucleus receiving converging auditory nerve fiber input. AB - The response of leaky integrate-and-fire neurons is analyzed for periodic inputs whose phases vary with their spatial location. The model gives the relationship between the spatial summation distance and the degree of phase locking of the output spikes (i.e., locking to the periodic stochastic inputs, measured by the synchronization index). The synaptic inputs are modeled as an inhomogeneous Poisson process, and the analysis is carried out in the Gaussian approximation. The model has been applied to globular bushy cells of the cochlear nucleus, which receive converging inputs from auditory nerve fibers that originate at neighboring sites in the cochlea. The model elucidates the roles played by spatial summation and coincidence detection, showing how synchronization decreases with an increase in both frequency and spatial spread of inputs. It also shows under what conditions an enhancement of synchronization of the output relative to the input takes place. PMID- 11932563 TI - An in vitro study of glenohumeral performance after suprascapular nerve entrapment. AB - PURPOSE: The functional performance of athletes with suprascapular nerve entrapment remains unaddressed biomechanically. The purpose of this study was to investigate the glenohumeral joint kinematics, stability, and characteristics of joint contact. METHODS: The investigation used an in vitro model simulating muscle palsy due to the suprascapular nerve entrapment. The experiment was performed in three clinically relevant positions: inferior, one by sulcus test; posterior, one with 90 degrees flexion and maximal internal rotation; and anterior, one with 90 degrees abduction and 90 degrees external rotation (which is critically involved in several sports activities). RESULTS: Simulated palsy due to suprascapular nerve entrapment had effects on glenohumeral joint stability and contact characteristics. Changes in the humeral head displacement and the glenohumeral joint contact characteristics were the most significant in the anterior direction when the arm in 90 degrees abduction and 90 degrees external rotation. CONCLUSION: Glenohumeral function is affected by muscle weakness at the position that is critical in sports activities. Because the infraspinatus supplies 90% of the external rotation power of the shoulder and the supraspinatus stabilizes the humeral head in the glenoid during elevation, residual weakness due to suprascapular nerve entrapment may preclude a safe return to athletics. PMID- 11932564 TI - Beta-blockers, exercise, and the immune system in men with coronary artery disease. AB - PURPOSE: The purpose of the present study was to examine the effect of aerobic exercise training on the immune system in coronary artery disease patients treated with beta-blockers. METHODS: Twenty-five patients (46.1 +/- 2 yr) treated with atenolol, a beta-blocker agent, for 3 months before exercising were divided randomly into two groups: 15 underwent an aerobic exercise training program for 12 wk at 65-70% of their work capacity, whereas the other 10 patients served as controls. RESULTS: After training, levels of CD4 and CD8 cells were significantly (P < 0.05) higher and concomitantly the CD4/CD8 ratio decreased significantly (P < 0.05) in the exercising group compared with the control group. CONCLUSIONS: Data suggest that coronary artery disease patients exercising aerobically at 65 70% of their work capacity gain a statistically significant higher lymphocyte T cell function as compared with their untrained counterparts. PMID- 11932565 TI - Cardiorespiratory fitness and stroke mortality in men. AB - PURPOSE: We examined the association between cardiorespiratory fitness and stroke mortality in men. METHODS: This is a prospective cohort study. We followed 16,878 men, ages 40-87 yr, who had a complete medical evaluation including a maximal treadmill exercise test and self-reported health habits. There were 32 stroke deaths during an average of 10 yr of follow-up (167,961 man-yr). RESULTS: After adjustment for age and examination year, there was an inverse association between cardiorespiratory fitness and stroke mortality (P = 0.005 for trend). This association remained after further adjustment for cigarette smoking, alcohol intake, body mass index, hypertension, diabetes mellitus, and parental history of coronary heart disease (P = 0.02 for trend). High-fit men (most fit 40%) had 68% (95% CI: 0.12, 0.82) and moderate-fit men had 63% (95% CI: 0.17, 0.83) lower risk of stroke mortality when compared with low-fit men (least fit 20%), respectively. CONCLUSIONS: Moderate and high levels of cardiorespiratory fitness were associated with lower risk of stroke mortality in men in the Aerobics Center Longitudinal study population. PMID- 11932566 TI - Exercise training in obesity lowers blood pressure independent of weight change. AB - PURPOSE: We used the rabbit model of obesity and exercise training to determine effects of exercise training during the development of obesity on resting blood pressure and heart rate, ventricular hypertrophy, blood volume, and hormonal profile. METHODS: Female New Zealand white rabbits were assigned to one of four groups: lean sedentary (L-S, N = 17), lean exercise-trained (L-EX, N = 16), obese sedentary (O-S, N = 18), and obese exercise-trained (O-EX, N = 15). Lean rabbits were fed a maintenance diet whereas obese rabbits were fed an ad libitum high fat (10% added fat) diet. Simultaneously, exercise-trained animals underwent a progressive treadmill exercise training protocol for 12 wk. After 12 wk of diet and exercise regimens, resting blood pressure and heart rate were measured from a central ear artery catheter. Ventricular hypertrophy was evaluated using wet ventricular weights. Blood volume was measured using the Evans blue dye procedure; hormonal profile was evaluated from arterial plasma/serum samples. RESULTS: After 12 wk, O-S and O-EX had similar body weights and similar percentage increases in body weight. Despite similar body weights, O-EX had an approximate 6-mm Hg lower mean blood pressure compared with the elevated pressure seen in O-S (P < or = 0.05). Obese rabbits had greater resting heart rate, plasma cholesterol and triglycerides, and plasma renin activity compared with lean rabbits, and these values were unaffected by exercise training. Plasma and blood volumes, as well as plasma insulin, cortisol, and aldosterone were unaffected by exercise training. CONCLUSION: These data suggest that exercise training, in the absence of differences in body weight, may be useful in the reduction of obesity induced hypertension but that other therapies may be needed in order to control other cardiovascular risk factors. PMID- 11932567 TI - Electro-membrane microcurrent therapy reduces signs and symptoms of muscle damage. AB - PURPOSE: Delayed onset muscle soreness (DOMS) occurs after unaccustomed physical activity or competitive sport, resulting in stiff, painful muscles with impaired function. Acustat electro-membrane microcurrent therapy has been used to treat postoperative pain and soft tissue injury; however, its efficacy in reducing symptoms of muscle damage is not known. METHODS: Thirty healthy men were recruited for a double-blind, placebo-controlled trial. The muscles of their nondominant arms were damaged using an eccentric-exercise protocol. Subjects were then randomly assigned to treatment with either Acustat or a matching placebo membrane for 96 h and monitored for a total of 168 h. RESULTS: Subjects in both groups experienced severe pain and swelling of the elbow flexors after the eccentric exercise. After 24 h, the elbow joint angle of the placebo group had increased significantly more than those in the Acustat group (13.7 +/- 8.9 degrees vs 7.5 +/- 5.5 degrees; placebo vs Acustat, P < 0.05), possibly as a consequence of the elbow flexor muscles shortening. For the first 48 h after exercise, maximum voluntary contraction of the elbow flexor muscles was significantly impaired in the placebo group by up to 25% (P < 0.05), whereas muscle function was unchanged in the Acustat group. Peak plasma creatine kinase activity was also lower in the Acustat group (peak = 777 +/- 1438 U.L-1) versus the placebo group (peak = 1918 +/- 2067 U.L-1; (P < 0.05). The membranes were well tolerated by the subjects in both groups without any adverse effects. CONCLUSION: These data show that treatment of muscle damage with Acustat electro membrane microcurrent therapy reduces the severity of the symptoms. The mechanisms of action are unknown but are likely related to maintenance of intracellular Ca2+ homeostasis after muscle damaging exercise. PMID- 11932568 TI - Leptin concentrations experience a delayed reduction after resistance exercise in men. AB - PURPOSE: Leptin is an important metabolic hormone providing the brain with information concerning energy balance. Most studies have reported that circulating leptin concentrations are unaltered by acute, moderate exercise. We hypothesized that these studies have been limited by short sampling schemes (<4 h) postexercise and may have missed a time-delayed reduction in circulating leptin concentrations. METHODS: Ten men (age = 21 +/- 1 yr, height = 177 +/- 2 cm, body mass = 79 +/- 3 kg, body fat = 11 +/- 1%BF, .VO(2max) = 51 +/- 1 mL.kg 1.min-1) completed an acute heavy-resistance exercise protocol (AHREP) (50 total sets comprised of the squat, bench press, leg press, and lat pull-down) from 1500 to 1700 h. Blood was sampled hourly postexercise until 0600 h the next morning and also during a time-matched control period. Leptin concentrations were measured by an immunoradiometric assay. Resting energy expenditure (REE) was measured via indirect calorimetry using a ventilated hood beginning approximately 0600 h after both overnight conditions. RESULTS: The estimated caloric expenditure from the AHREP was 856 +/- 114 kcal. No significant differences (P > 0.05) between the control and exercise conditions were observed for serum leptin concentrations until 9 h postexercise. Significant interaction effects (P < 0.05) indicated lower serum leptin concentrations postexercise at hours 9 (2.9 vs 2.2 ng.mL-1), 10 (2.7 vs 2.0 ng.mL-1), 12 (2.5 vs 1.8 ng.mL-1), and 13 (2.6 vs 1.8 ng.mL-1). This delayed reduction was accompanied by a 12% elevation (P < 0.05) in morning-after REE (0.25 +/- 0.02 vs 0.28 +/- 0.02 L.min-1). CONCLUSION: Leptin concentrations experience a delayed ( approximately 9 h) reduction in the systemic circulation after acute resistance exercise. This decline is likely associated with the disruption in metabolic homeostasis created by the high intensity, long-duration, energy expenditure and subsequent excess post oxygen consumption from the AHREP and is not due to losses in fat mass. PMID- 11932569 TI - Effect of sodium bicarbonate on muscle metabolism during intense endurance cycling. AB - INTRODUCTION: Sodium bicarbonate (NaHCO3) ingestion has been shown to increase both muscle glycogenolysis and glycolysis during brief submaximal exercise. These changes may be detrimental to performance during more prolonged, exhaustive exercise. This study examined the effect of NaHCO3 ingestion on muscle metabolism and performance during intense endurance exercise of approximately 60 min in seven endurance-trained men. METHODS: Subjects ingested 0.3 g.kg-1 body mass of either NaHCO3 or CaCO3 (CON) 2 h before performing 30 min of cycling exercise at 77 +/- 1% .VO(2peak) followed by completion of 469 +/- 21 kJ as quickly as possible (approximately 30 min, approximately 80% .VO(2peak)). RESULTS: Immediately before, and throughout exercise, arterialized-venous plasma HCO3- concentrations were higher (P < 0.05) whereas plasma and muscle H+ concentrations were lower (P < 0.05) in NaHCO3 compared with CON. Blood lactate concentrations were higher (P < 0.05) during exercise in NaHCO3, but there was no difference between trials in muscle glycogen utilization or muscle lactate content during exercise. Reductions in PCr and ATP and increases in muscle Cr during exercise were also unaffected by NaHCO3 ingestion. Accordingly, exercise performance time was not different between treatments. CONCLUSION: NaHCO3 ingestion resulted in a small muscle alkalosis but had no effect on muscle metabolism or intense endurance exercise performance in well-trained men. PMID- 11932570 TI - Modeling of end-tidal and arterial PCO2 gradient: comparison with experimental data. AB - PURPOSE: The aim of this study was to test whether a tidally ventilated homogeneous lung model can correctly describe arterial and end-tidal gas partial pressures and thus the difference in end-tidal and arterial gas partial pressures at rest and during exercise. METHODS: The implemented mathematical modeling described variations during the breathing cycle in CO2 and O2 fractions, alveolar volume, and pulmonary capillary gas exchange. Experimental data were obtained from measurements performed by 17 healthy subjects at rest and during 40, 50, 65, and 75% exercise .VO(2max) on a cycle ergometer. VO2, VCO2, and PET,CO2 were continuously measured using the MedGraphics CPX/D gas exchange system. Arterial gases were measured in brachial artery blood samples drawn simultaneously with gas exchange. Cardiac output was measured using the CO2 rebreathing method corrected by the blood sample data. The model was driven using experimental data for ventilation, VO2, VCO2, and cardiac output. RESULTS: The mean difference and the upper and lower limits of agreement between measured and simulated data were 0.004, +0.84, and -0.84 Torr for Pa,CO2; -0.06, +0.64, and -0.76 Torr for Pa,O2; 1.96, +2.84, and -6.76 Torr for PET,CO2; and +7.20, +25.80, and -11.40 Torr for PET,O2. Actual PET,CO2-Pa,CO2 difference increased significantly with workload (P < 0.0001) from 0.3 +/- 3 Torr at rest to 4.7 +/- 2.5 Torr at 75% .VO(2max). Model simulated PET,CO2-Pa,CO2 difference also increased significantly with exercise (P < 0.0001) from 0.7 +/- 1.7 Torr at rest to 9.1 +/- 3.4 Torr at 75% .VO(2max). CONCLUSION: The lung model described actual arterial CO2 partial pressures better than variations in end-tidal CO2 partial pressures and thus better than the gradient in end-tidal arterial CO2 partial pressures. PMID- 11932571 TI - Hyperbaric oxygen increases the contractile function of regenerating rat slow muscles. AB - Human trials of hyperbaric oxygen (HBO) treatment of sports-related muscle injuries are equivocal. Although most human skeletal muscles are composed of mixed muscle fiber types, it is unclear whether HBO affects fiber types differently. PURPOSE: We tested the hypothesis that HBO can enhance the functional properties of regenerating rat soleus muscles that are composed predominantly of slow fibers. METHODS: After intramuscular injection of bupivacaine hydrochloride to induce the degeneration of all fibers within the soleus muscle, treated rats received daily HBO treatment at 3 atmospheres absolute. RESULTS: In untreated rats, injured muscles demonstrated a reduced force-producing capacity (control soleus vs injured soleus, 220.3 +/- 2.5 vs 157.6 +/- 3.3 kN.m(-2) at 25 d postinjury, respectively, P < 0.05) and contained smaller regenerating muscle fibers than uninjured soleus muscles (fiber cross sectional area in control soleus vs injured soleus, 2289 +/- 164 vs 1154 +/- 92 microm 2 at 25 d postinjury, respectively, P < 0.05). The regenerating soleus muscles of HBO-treated rats demonstrated a greater force-producing capacity as a percentage of contralateral control muscles than the regenerating muscles from untreated rats at 14 d postinjury (regenerating HBO-soleus peak tension and untreated soleus peak tension, 42.9 +/- 1.9 and 35.8 +/- 3.9% of contralateral control muscles, respectively, P < 0.05), but no effect of treatment was observed at 25 d postinjury. CONCLUSION: HBO enhanced the contractile properties of regenerating rat soleus muscles after myotoxic injury, but this improvement was not sustained for the duration of the regenerative process. The data indicate that the outcome of HBO treatment of a muscle injury may be influenced by the fiber type composition of the injured muscle. PMID- 11932572 TI - Left ventricular response to dynamic exercise in young cyclists. AB - PURPOSE: The purpose of this study was to compare cardiac physiological and dimensional responses to exercise in highly trained young male cyclists (mean age 13.7 +/- 1.0 yr) with those of nontrained boys. METHODS: Ventricular systolic and diastolic dimensions were measured by two-dimensional echocardiography, and stroke volume was estimated by Doppler echocardiography during a progressive maximal upright cycle test. RESULTS: At rest, the cyclists demonstrated larger left ventricular dimensions relative to body size than the nonathletes. Maximal stroke index and cardiac index were significantly greater in the cyclists. The pattern of stroke volume response to exercise was similar in the two groups, with an early rise and then plateau to exhaustion. Left ventricular diastolic dimension increased slightly at onset of exercise and then gradually declined as workload increased in both groups. CONCLUSION: Factors responsible for the greater maximal stroke volume in young endurance athletes involve those variables that contribute to resting left ventricular diastolic filling (preload). PMID- 11932573 TI - Chronic clenbuterol administration negatively alters cardiac function. AB - PURPOSE: Chronic administration of pharmacological levels of beta2-agonists have been shown to have toxic effects on the heart; however, no data exist on cardiac function after chronic clenbuterol administration. The purpose of this study was to examine the effect of therapeutic levels of clenbuterol on cardiac performance. METHODS: Twenty unfit Standardbred mares were divided into four experimental groups: clenbuterol (2.4 microg.kg(-1) twice daily 5 d.wk(-1)) plus exercise (20 min at 50% .VO(2max)) (CLENEX; N = 6), clenbuterol (CLEN; N = 6), exercise (EX; N = 4), and control (CON; N = 4). M-mode and two-dimensional echocardiography (2.5-MHz sector scanner transducer) were used to measure cardiac size and function before and immediately after an incremental exercise test, before and after 8 wk of drug and/or exercise treatments. RESULTS: After treatment, CLENEX and CLEN demonstrated significantly higher left ventricular internal dimension (LVD) at end diastole (+23.7 +/- 4.8%; +25.6 +/- 4.1%), LVD at end systole (+29.2 +/- 8.7%; +40.1 +/- 7.9%), interventricular septal wall thickness (IVS) at end diastole (+28.9 +/- 11.0%; +30.7 +/- 7.0%), IVS at end systole (+29.2 +/- 8.7%; +40.1 +/- 7.9%), and left ventricular posterior wall systolic thickness (+43.1 +/- 14.%; +45.8 +/- 14.1%). CLENEX and CLEN had significantly increased aortic root dimensions (+29.9 +/- 6.1%; +24.0 +/- 1.7%), suggesting increased risk of aortic rupture. CONCLUSION: Taken together, these data indicate that chronic clenbuterol administration may negatively alter cardiac function. PMID- 11932574 TI - The cardiospecificity of the third-generation cTnT assay after exercise-induced muscle damage. AB - PURPOSE: The purpose of the present study was to examine the cardiospecificity of cTnI and the new third-generation cTnT assay, in the presence of exercise-induced muscle damage in highly trained individuals, and to examine the impact of a maximal-ramping treadmill test on cardiac function. METHODS: Eight highly trained male triathletes (mean +/- SD; age: 29 +/- 9 yr; height: 1.79 +/- 0.10 m; body mass: 77 +/- 10 kg; .VO(2max): 67.4 +/- 6.3 mL.kg(-1).min(-1)) completed two bouts of exercise. On the first occasion, subjects completed a maximal-ramping treadmill test. On a separate occasion, the subjects completed 30 min of downhill running (15% gradient) at a speed equivalent to 70% of maximal running velocity attained during the maximal-ramping treadmill test. All subjects were assessed using ECG, echocardiography, and blood analysis. Measurements were taken at rest, immediately after, and 48 h postexercise for each bout of exercise. Echocardiographic analysis was used to determine left ventricular systolic and diastolic function. Blood samples were analyzed for markers of myocyte damage. RESULTS: Echocardiographic results indicated normal left ventricular function before and after both exercise bouts. Total CK and CKMB were significantly elevated 48 h after the downhill run. cTnT and cTnI were not elevated at any stage of the study. CONCLUSIONS: Neither the maximal-ramping treadmill test nor the 30-min downhill run produced cardiac dysfunction or myocardial damage in young, healthy trained subjects. The elevated total CK and CKMB within the downhill study are noncardiac in origin as demonstrated by the lack of cTnT and cTnI. The cTnI and new third-generation cTnT assays may be used to detect cardiac damage in the presence of elevated total CK and CKMB associated with exercise induced skeletal muscle damage. PMID- 11932575 TI - Muscle fiber type distribution and nonlinear .VO(2)-power output relationship in cycling. AB - PURPOSE: We examined whether reported deviations from linearity of the oxygen uptake (.VO(2))-to-power output (W) relationship during intense cycling exercise correlated with the percentage Type II fibers in the exercising muscle. METHODS: Twelve trained young men with known fiber type distribution in the vastus lateralis muscle performed step-increment exercise (40 W.3 min(-1)) to exhaustion. RESULTS: .VO(2) increased linearly with W up to about 50% .VO(2max) with a regression equation of .VO(2) (mL.min-1) = 661 + 9.73 W and a correlation coefficient (r) of 1.000. Subsequent .VO(2) values were all greater than corresponding linear estimates (P < 0.001 or 0.0001). Peak exercise excess .VO(2) (measured minus estimated .VO(2) assuming linearity) averaged (SD) 434 (192) mL O(2).min-1 or 10.3 (4.7) % .VO(2max). A comprehensive curvilinearity index defined as the sum of measured minus estimated .VO(2) at the four highest completed exercise trials averaged 973 (460) mL O(2).min-1 or 21.5 (9.4) % .VO(2max). Correlations between percentage Type II fibers and either of the two expressions of curvilinearity were nonsignificant. Delta [H+] (arterialized capillary blood) from basal level to peak exercise correlated with the submaximal curvilinearity index (r = 0.59-0.64; P < 0.05) but not with peak excess .VO(2). There was a trend toward a correlation between delta La and curvilinearity index in % .VO(2max)(r = 0.52; P < 0.10) but not with any of the other curvilinearity expressions. The relative ventilatory activity expressed as .V(E)-to-.VO(2) ratio tended to correlate with peak excess .VO(2) (P < 0.10) but not with curvilinearity index. Signals from motion sensors indicate that coactivation of upper-body musculature coincided with deviation from linearity in the .VO(2)-W relationship. CONCLUSION: VO2 during step-increment cycling increases linearly with power output up to about 50% .VO(2max)and then curvilinearly. The degree of curvilinearity is not related to muscle fiber type distribution in the vastus lateralis, and only marginally and insignificantly related (P < 0.10) to the relative degree of hyperventilation or to lactate response. Acidosis, on the other hand, correlated significantly with curvilinearity index. The inclusion of isometrically working, upper-body muscular groups during high-intensity cycling may also contribute to the overshoot in oxygen cost. PMID- 11932576 TI - A kinematic analysis of high-speed treadmill sprinting over a range of velocities. AB - INTRODUCTION: The purpose of this study was to measure changes in stride characteristics and lower-extremity kinematics of the hip and knee as a function of increasing treadmill velocity, at velocities ranging from submaximal to near maximal. METHODS: Six power/speed athletes experienced at sprinting on a treadmill performed trials at 70%, 80%, 90%, and 95% of their previous individual maximum velocity, with video data collected in the sagittal view at 60 Hz. RESULTS: Significant differences were seen in stride frequency (70%, 80%, P < 0.01; 90%, P < 0.05), stance time (70%, 80%, P < 0.01; 90%, P < 0.05) flight time (70%, P < 0.01; 80%, P < 0.05), hip flexion angle (70%, P < 0.01), hip flexion angular velocity (70%, P < 0.01), hip extension angular velocity (70%, 80%, P < 0.01), knee flexion angular velocity (70%, 80%, P < 0.01), and knee extension angular velocity (70%, P < 0.01), as compared with the near maximum (95%) velocity. Coefficient of variation (CV) values showed that the positional variables at the hip and knee were more variable at faster test conditions, indicating that kinematic changes occur as a function of increased treadmill velocity. CONCLUSIONS: The results indicated that at slower velocities, there were differences in the stride characteristics and lower-extremity kinematics while sprinting on a treadmill. As the velocity approached near maximum mechanical breakdown was seen, suggesting that velocities greater than 90% should be used selectively during treadmill training. PMID- 11932577 TI - Force deficits by stretches of activated muscles with constant or increasing velocity. AB - PURPOSE: Force deficits produced by constant (CV) versus increasing velocity (IV) stretches of rat plantar flexor muscles at low and high levels of nerve activation were studied. METHODS: Twenty repeated stretches were imposed on isometric contractions by ankle rotation from 90 degrees to 40 degrees at 300 degrees.s(-1) and at 3000 degrees.s(-2) during 80-Hz (CV80 and IV80) and 20-Hz stimulation (CV20 and IV20). Rest periods between contractions were 3 min. Isometric and peak stretch forces during the stretch protocols and force frequency relationships before and 1 h after the stretch protocols were measured. RESULTS: Peak stretch forces were similar for IV80-CV80 and for IV20-CV20 rats but were lower for IV20-CV20 than for IV80-CV80 rats throughout the stretch protocol. At the end of the stretch protocol, isometric force deficits were similar for IV80 (49.9 +/- 2.1%) and CV80 (54.5 +/- 2.5%) and for IV20 (16.4 +/- 2.8%) and CV20 (15.8 +/- 1.9%) but lower for IV20-CV20 rats. In contrast, for all groups, deficits in peak stretch force were similar at the end of the stretch protocol (IV80: 35.0 +/- 1.8%, CV80: 32.3 +/- 2.2%, IV20: 26.8 +/- 3.6%, CV20: 28.0 +/- 2.0%). After 1 h, isometric force deficits were similar for either IV80 CV80 or IV20-CV20 at 5, 10, 20, 40, 60, and 80 Hz stimulation but were lower for IV20-CV20. CONCLUSIONS: Variation in velocity of ankle rotation with similar peak stretch forces did not influence the amount of stretch-induced force deficits. High peak stretch forces produced greater isometric force deficits than low peak stretch forces, but the relative loss in peak stretch force was not force dependent. Different mechanisms may account for isometric force deficits and peak stretch force deficits caused by repeated stretches of activated skeletal muscles. PMID- 11932578 TI - Mid-femur geometry and biomechanical properties in 15- to 18-yr-old female athletes. AB - PURPOSE: Right-leg mid-femur geometry and biomechanical indices of bone strength were compared among elite cyclists (CYC), runners (RUN), swimmers (SWIM), triathletes (TRI), and controls (C)-10 subjects per group. METHODS: Bone cross sectional areas (CSA), volumes (Vol), and cross-sectional moments of inertia (CSMI) were assessed by magnetic resonance imaging (MRI), and cortical volumetric bone density (volBMD) was determined as the quotient of DXA-derived bone mineral content (BMC) and MRI-derived cortical bone volume. Bone strength index (BSI) was calculated as the product of cortical volBMD and CSMI. RESULTS: RUN had higher (P < 0.05) size- (femur length and body mass) adjusted (ANCOVA) cortical CSA than C, SWIM, and CYC; and higher size, age, and years of sport-specific training- (YST) adjusted cortical CSA than SWIM and CYC. TRI had higher (P < 0.05) size-adjusted CSA than SWIM. SWIM and CYC had significantly larger (P < 0.05) size-adjusted medullary cavity CSA than RUN and TRI, and the difference between CYC and RUN persisted after additional adjustment for age and YST. RUN had significantly (P < 0.05) greater size-adjusted CSMI and BSI than C, SWIM, and CYC; and higher size, age, and YST-adjusted CSMI and BSI than SWIM and CYC. Mid-femur areal bone mineral density (BMD) was significantly (P < 0.05) higher for RUN compared with CYC only, but there were no other differences among groups for BMC, bone volumes, or volumetric total or cortical BMD. CONCLUSIONS: Running, a weight-bearing exercise, is associated with more favorable geometric and biomechanical characteristics in relation to bone strength, compared with the weight supported activities of swimming and cycling. Differences may reflect skeletal adaptations to the specific mechanical-loading patterns inherent in these sports. PMID- 11932579 TI - An electromyographic analysis of sumo and conventional style deadlifts. AB - PURPOSE: Strength athletes often employ the deadlift in their training or rehabilitation regimens. The purpose of this study was to compare muscle activity between sumo and conventional style deadlifts, and between belt and no-belt conditions. METHODS: Six cameras collected 60-Hz video data and 960-Hz electromyographic data from 13 collegiate football players who performed sumo and conventional deadlifts with and without a lifting belt, employing a 12-RM intensity. Variables measured were knee angles and EMG measurements from 16 muscles. Muscle activity were averaged and compared within three 30-degree knee angle intervals from 90 to 0 degrees during the ascent, and three 30-degree knee angle intervals from 0 to 90 degrees during the descent. RESULTS: Overall EMG activity from the vastus medialis, vastus lateralis, and tibialis anterior were significantly greater in the sumo deadlift, whereas overall EMG activity from the medial gastrocnemius was significantly greater in the conventional deadlift. Compared with the no-belt condition, the belt condition produced significantly greater rectus abdominis activity and significantly less external oblique activity. For most muscles, EMG activity was significantly greater in the knee extending intervals compared with the corresponding knee flexing intervals. Quadriceps, tibialis anterior, hip adductor, gluteus maximus, L3 and T12 paraspinal, and middle trapezius activity were significantly greater in higher knee flexion intervals compared with lower knee flexion intervals, whereas hamstrings, gastrocnemius, and upper trapezius activity were greater in lower knee flexion intervals compared with higher knee flexion intervals. CONCLUSIONS: Athletes may choose to employ either the sumo or conventional deadlift style, depending on which muscles are considered most important according to their training protocols. Moderate to high co-contractions from the quadriceps, hamstrings, and gastrocnemius imply that the deadlift may be an effective closed kinetic chain exercise for strength athletes to employ during knee rehabilitation. PMID- 11932580 TI - An assessment of maturity from anthropometric measurements. AB - PURPOSE: The range of variability between individuals of the same chronological age (CA) in somatic and biological maturity is large and especially accentuated around the adolescent growth spurt. Maturity assessment is an important consideration when dealing with adolescents, from both a research perspective and youth sports stratification. A noninvasive, practical method predicting years from peak height velocity (a maturity offset value) by using anthropometric variables is developed in one sample and cross-validated in two different samples. METHODS: Gender specific multiple regression equations were calculated on a sample of 152 Canadian children aged 8-16 yr (79 boys; 73 girls) who were followed through adolescence from 1991 to 1997. The equations included three somatic dimensions (height, sitting height, and leg length), CA, and their interactions. The equations were cross-validated on a combined sample of Canadian (71 boys, 40 girls measured from 1964 through 1973) and Flemish children (50 boys, 48 girls measured from 1985 through 1999). RESULTS: The coefficient of determination (R2) for the boys' model was 0.92 and for the girls' model 0.91; the SEEs were 0.49 and 0.50, respectively. Mean difference between actual and predicted maturity offset for the verification samples was 0.24 (SD 0.65) yr in boys and 0.001 (SD 0.68) yr in girls. CONCLUSION: Although the cross-validation meets statistical standards for acceptance, caution is warranted with regard to implementation. It is recommended that maturity offset be considered as a categorical rather than a continuous assessment. Nevertheless, the equations presented are a reliable, noninvasive and a practical solution for the measure of biological maturity for matching adolescent athletes PMID- 11932581 TI - Heart rate, oxygen uptake, and energy cost of ascending and descending the stairs. AB - PURPOSE: This study describes the heart rate and oxygen uptake responses during, and the intensity and caloric cost of, ascending and descending a public-access staircase. METHODS: Subjects were initially assessed for their maximum oxygen uptake and heart rate on a treadmill in the laboratory. For field measurements, subjects ascended (N = 103) and descended (N = 49) 11 stories of 180 steps, each step of 15 cm in height, for a total vertical displacement of 27.0 m. RESULTS: The mean oxygen uptake and heart rate during the last 30 s of ascending were 33.5 +/- 4.8 mL.kg(-1).min(-1) and 159 +/- 15 beats.min(-1), respectively. During the descent, oxygen uptake and heart rate during the last 30 s of the climb were 17.0 +/- 3.8 mL.kg(-1).min(-1) and 107 +/- 18 beats.min(-1), respectively. The estimated gross energy expended during ascending and descending were 19.7 and 9.0 kcal, or equivalent to an intensity of 9.6 and 4.9 metabolic equivalents (METs), respectively (or 10.2 and 5.2 kcal.min(-1), respectively). The caloric cost of stepping up and down a step was calculated to be 0.11 and 0.05 kcal, respectively. CONCLUSION: Stair-climbing exercise using a local public-access staircase met the minimum requirements for cardiorespiratory benefits and can therefore be considered a viable exercise for most people and suitable for promotion of physical activity. PMID- 11932582 TI - Aerobic training and cardiovascular responses at rest and during exercise in older men and women. AB - PURPOSE: The effects of an intense 8-wk aerobic training program on cardiovascular responses at rest and during exercise, including heart rate variability (HRV) as an expression of autonomic modulation, were evaluated in subjects over 70 yr (mean: 73.9 +/- 3.5 yr). METHODS: Before and after training in 7 men and 8 women: a) heart rate (HR), blood pressures (BPs), pulse pressure (PP), and oxygen uptake were measured at rest, during, and after exhausting incremental exercise; b) HRV power spectra were calculated at rest in supine and sitting, and during and after two submaximal constant loads (5 min). Power in low frequency (LF, 0.04-0.15 Hz) and high-frequency (HF, >0.15 Hz) bands were expressed as a percent of total power minus power < 0.04 Hz. RESULTS: After training: a) at rest HR and HRV parameters (in both body positions) were unchanged, whereas BPs decreased; b) peak cycle resistance and oxygen consumption increased by 25% and 18%, respectively, but no change in maximal HR and BPs were found; c) during submaximal loads HR was unchanged at the same metabolic demand, whereas SBP and DBP were lower than before at low loads whereas PP was unchanged. LF power decreased and HF increased at oxygen uptakes above about 0.7 L.min-1 similarly before and after training; and d) recovery of all parameters was similar to pretraining and complete after 10 min CONCLUSIONS: The increase in exercise capacity without changes in cardiovascular parameters suggests that 8 wk of aerobic training augmented peripheral gas exchange but not delivery to muscle. The lack of effect on HRV indicates that the improvements in aerobic power and cardiac autonomic modulation, at least in subjects over 70 yr, are dissociated. Moreover, the metabolic demand seems to be the main factor for the changes in HRV power spectra that occur during exercise. PMID- 11932583 TI - The relationship between power and the time to achieve .VO(2max). AB - PURPOSE: The severe exercise intensity domain may be defined as that range of work rates over which .VO(2max) can be elicited during constant-load exercise. The purpose of this study was to help characterize the .VO(2) response within this domain. METHODS: Eleven participants performed cycle ergometer exercise tests to fatigue at several discrete work rates between 95% and 135% of the maximum power (P(max)) achieved during an incremental exercise test. RESULTS: As previously demonstrated, the relationship between power and time to fatigue was hyperbolic. The asymptote of power (critical power, P(critical)) was 198 +/- 44 W. The rapidity of the .VO(2) response increased systematically at higher work rates such that the relationship between power and time to .VO(2max) was also well fit by a hyperbola. The power asymptote of this relationship (196 +/- 42 W) was not different from P(critical)(P > 0.05). The two hyperbolic relationships converged at 342 +/- 70 W (136% P(max)). CONCLUSION: These data suggest that, for this population of male and female university students, the upper boundary of the severe exercise intensity domain is approximately 136% P(max). This upper boundary is the highest work rate for which exercise duration is prolonged sufficiently (in this study, 136 +/- 17 s) to allow .VO(2) to rise to its maximal value. The lower boundary for severe exercise is just above P(critical), which is the highest work rate that is sustainable for a prolonged duration and that will not elicit .VO(2max). PMID- 11932584 TI - Effects of resistance exercise bouts of different intensities but equal work on EPOC. AB - PURPOSE: To compare the effect of low- and high-intensity resistance exercise of equal work output, on exercise and excess postexercise oxygen consumption (EPOC). METHODS: Fourteen female subjects performed a no-exercise baseline control (CN), and nine exercises for two sets of 15 repetitions at 45% of their 8-RM during one session (LO) and two sets of 8 repetitions at 85% of their 8-RM during another session (HI). Measures for all three sessions included: heart rate (HR) and blood lactate (La) preexercise, immediately postexercise and 20 min, 60 min, and 120 min postexercise; and ventilation volume (VE), oxygen consumption (VO(2)), and respiratory exchange ratio (RER) during exercise and at intervals 0-20 min, 45-60 min, and 105-120 min postexercise. RESULTS: Exercise .VO(2) was not significantly different between HI and LO, but VE, [La], and HR were significantly greater for HI compared with LO. Exercise RER for HI (1.07 +/- 0.03 and LO (1.05 +/- 0.02) were significantly higher than CN (0.86 +/- 0.02), but there were no differences among conditions postexercise. EPOC was greater for HI compared with low at 0-20 min (HI,1.72 +/- 0.70 LO(2); LO, 0.9 +/- 0.65, LO(2)), 45-60 min (HI, 0.35 +/- 0.25 LO(2); LO, 0.14 +/- 0.19 LO2), and 105-120 min (HI, 0.22 +/- 0.22 LO(2); LO, 0.05 +/- 0.11, LO(2)). CONCLUSION: These data indicate that for resistance exercise bouts with an equated work volume, high-intensity exercise (85% 8-RM) will produce similar exercise oxygen consumption, with a greater EPOC magnitude and volume than low-intensity exercise (45% 8-RM). PMID- 11932585 TI - Carbohydrates and physical/mental performance during intermittent exercise to fatigue. AB - PURPOSE: This study was designed to examine the effects of carbohydrate electrolyte ingestion on physical and mental function associated with the performance of intermittent high-intensity (IHI) exercise similar to many common competitive sporting events. METHODS: Physically active men (N = 5) and women (N = 5), experienced in competitive soccer or basketball, completed three practice sessions and two experimental trials of an IHI shuttle running protocol designed to closely stimulate the demands of an actual competitive sporting event such as basketball. The experimental trials consisted of four 15-min quarters (QTR) of intermittent shuttle running at various percentages of .VO(2max) (walking, jogging, running, sprinting and jumping), separated by a 20-min halftime rest period (HALF) and followed by a shuttle run to fatigue. Various tests of physical and mental function (shuttle run to fatigue, 20-m maximal sprint, 10-repetition maximal vertical jumping, whole body motor skill test (MS-Test), profile of mood states (POMS), and Stroop Color-Word Test) were performed throughout the experimental trial. Carbohydrate-electrolyte (CHO) or placebo (P) drinks were consumed before exercise (5 mL.kg(-1); 6% solution) and at halftime (5 mL.kg(-1); 18% solution). Smaller volumes (3 mL.kg(-1); 6% solution) were given after QTR-1, HALF, QTR-3, and QTR-4. RESULTS: CHO ingestion resulted in a 37% longer run time to fatigue and faster 20-m sprint time during QTR-4 (P < 0.05). MS-Test performance was also improved during the latter stages of exercise along with self-reported perceptions of fatigue (subscale of POMS) (P < 0.05) in CHO versus P. CONCLUSION: These results suggest a beneficial role of carbohydrate electrolyte ingestion on physical and mental function during intermittent exercise similar to that of many competitive team sports. PMID- 11932587 TI - Balanced auricular reconstruction in dystopic microtia with the presence of the external auditory canal. AB - This article presents a new repositioning method in dystopic microtia (low-set microtia, anteriorly tilted microtia, or both) with the presence of the external auditory canal. In the case of low-set malformations, the dystopic auricular canal complex was freed from adjacent bony structure, shifted upward, and anchored to the thick periosteum of the suprameatal triangle or the adjacent superior portion of the temporal bone with nonabsorbable sutures. When the auricular vestige was large and dystopia was severe, the complex was shifted with attachment of the temporoparietal fascia on its cranial part. Meanwhile, when the auricular vestige was small and dystopia was mild, the complex was shifted without attachment of the temporoparietal fascia. Then, the ear framework fabricated with autogenous costal cartilage was positioned and wrapped with the temporoparietal fascial flap. In the case of anteriorly tilted malformations, the dystopic complex was freed, shifted posteriorly, and anchored to the periosteum of the suprameatal triangle. Preauricular dead space, resulting from shifting the complex, was obliterated with pieces of costal cartilage. Simultaneously, the ear framework was placed and wrapped with the temporoparietal fascial flap. No skin necrosis of the shifted complexes occurred in any of the cases. In one case, the facial nerve was transected during dissection and reanastomosed. Upward repositioning distances in low-set microtias were between 1 and 3.5 cm. Posteriorly repositioning distances in anteriorly tilted microtias were 2 and 3 cm. Thirteen patients with low-set malformations, two patients with anteriorly tilted malformations, and three patients with low-set and anteriorly tilted malformations underwent reconstructive operations. The new repositioning method is relatively simple, safe, and effective. PMID- 11932589 TI - Congenital nasal pyriform aperture stenosis: diagnosis and treatment. AB - Congenital nasal pyriform aperture stenosis is an unusual form of nasal airway obstruction in the neonate. Pediatric plastic surgeons are often involved in the management of these children and should recognize this condition and know the treatment options. Fifteen cases of children with congenital nasal pyriform aperture stenosis were reviewed for presentation of the disorder, management, and effectiveness of treatment, making it the largest series to date. There were nine male patients and six female patients in the series. They all experienced varying degrees of nasal obstruction at birth and were managed on the basis of the severity of their symptoms. Twelve patients were treated surgically in the first year of life, with a mean age at operation of 97 days (range, 3 to 362 days). Two patients required surgical intervention during their teenage years (age, 14 and 18 years) because of persistent symptoms, and one patient (age, 2 years) with mild symptoms was managed medically. Associated craniofacial anomalies were present in six cases (40 percent). Surgical enlargement of the pyriform aperture was successfully performed through an upper buccal sulcus incision in 14 patients. Preoperative symptoms of upper airway obstruction were improved in all patients at an average follow-up of 2.4 years (range, 1 month to 5 years). Congenital nasal pyriform aperture stenosis varies in presentation and severity, occurring either as an isolated congenital anomaly or in association with developmental craniofacial anomalies. It can be effectively managed by surgical enlargement of the pyriform aperture without significant recurrence or long-term morbidity. PMID- 11932591 TI - The course of the inferior alveolar nerve in craniofacial microsomia: virtual dissection using three-dimensional computed tomography image analysis. AB - Computer-assisted medical imaging was used to locate the mandibular foramen and the portion of the inferior alveolar nerve canal at the level of the angle of the mandible in 19 hemimandibles of patients with craniofacial microsomia. The distance from each of these two points to the borders of the mandible was measured. The ratios of these distances to the height, anteroposterior, and buccolingual extents of the mandibular ramus were calculated. These ratios were compared among affected hemimandibles (Pruzansky classification I, n = 4; and Pruzansky classification II, n = 10), unaffected hemimandibles (n = 9), and the hemimandibles of patients with nonsyndromic, sutural synostosis (n = 7). Comparison of mean distance ratios showed that the mandibular foramen was located significantly more proximally in the affected cohort than in either the unaffected or synostosis control groups. The mandibular foramen was also located significantly closer to the buccal cortex of the mandible in the affected cohort when compared with unaffected patients. The distance from the mandibular foramen to the anterior or posterior tables of the mandible divided by the total anteroposterior distance did not vary among the three groups studied. The distance ratios between the inferior alveolar nerve canal at the level of the angle of the mandible and the mandibular borders also did not vary significantly among the groups studied. An inferior alveolar nerve canal could not be identified in any patient with Pruzansky grade III mandibular deficiency. This quantitative, three-dimensional description of points along the proximal path of the inferior alveolar nerve canal in patients with craniofacial microsomia provides useful information to assist the surgeon during osteotomy planning and may help in avoiding injury to the nerve at the time of surgery. PMID- 11932592 TI - Analysis of pharyngocutaneous fistula following free jejunal transfer for total laryngopharyngectomy. AB - The development of a pharyngocutaneous fistula is the most common and troublesome complication in the early postoperative period following free jejunal transfer for total laryngopharyngectomy. However, many aspects of this complication remain unclear. In this study, the authors analyzed their experience with the pharyngocutaneous fistula formation following free jejunal transfers to evaluate its clinical behavior, determine the significance of the anastomotic technique used, and evaluate the role of preoperative radiation therapy on its formation and management. Of 168 patients who underwent free jejunal transfers following total laryngopharyngectomy at the authors' institution between July of 1988 and March of 2000, 23 patients (13.7 percent) with postoperative fistulas were identified. The mean onset of fistula formation was 16 days. Of the 23 fistulas, 13 (56.5 percent) occurred at the proximal and 10 (43.5 percent) at the distal anastomoses. Whereas the majority of the proximal fistulas (69.2 percent) developed near the mesenteric side of the jejunal flap, most of the distal fistulas (90 percent) were located anteriorly. The incidence of proximal fistula formation was higher in patients with a single-layer repair than in patients with a two-layer repair of a proximal anastomosis (80 percent versus 38.5 percent, p = 0.09). The incidence of fistula formation was greater in patients who received preoperative radiation therapy than in those who did not (16.3 percent versus 11.4 percent, p = 0.36). In addition, whereas a majority of fistulas (80 percent) occurred at the proximal anastomosis in patients who did not receive preoperative radiation therapy, most fistulas (61.5 percent) occurred at the distal anastomosis in patients who did receive radiation therapy (p = 0.09). The fistulas closed spontaneously in 15 patients (65 percent). On average, spontaneous closure occurred in 7.4 weeks. Proximal fistulas had a significantly higher rate of spontaneous closure compared with distal fistulas (85 percent versus 40 percent, p = 0.04). The rate of spontaneous fistula closure was higher in patients who had not received preoperative radiation therapy than in those who had (90 percent versus 46 percent, p = 0.07). Surgical closure of the fistula was required in five patients. The fistulas were not repaired in three patients because of recurrent tumor. Twenty patients (87 percent) resumed oral feeding after the closure of the fistula, with 17 (85 percent) of 20 patients tolerating a regular diet and three (15 percent) of 20 a liquid diet only.In conclusion, most fistulas occur at the proximal anastomosis and near the mesenteric side of the jejunal flap, and the use of a two-layer anastomotic technique seems to be associated with a lower incidence of fistula formation at the proximal suture line. Most fistulas close spontaneously, especially ones that occur proximally. Preoperative radiotherapy does seem to increase the risk of fistula formation, especially at the distal anastomotic site and make subsequent resolution of the fistulas more difficult. Most patients are able to resume oral feeding once the fistula is closed. PMID- 11932593 TI - Reconstruction of facial defects with superficial temporal artery island flaps: a donor site with various alternatives. AB - Color and texture match is crucial in reconstruction of facial tissue defects. Between March of 1997 and July of 2000, island flaps based on the parietal, anterofrontal, centrofrontal, posterofrontal, and superior auricular branches of the superficial temporal artery were used in the reconstruction of tissue defects localized on different regions of the face in 28 patients. According to the size and the location of the defect, the flap was selected. There were 15 male patients and 13 female patients, with ages ranging between 19 and 74 years. In six of the flaps, venous congestion was observed. Because of the elevation of the eyebrow on the flap side, three patients required a sling to the opposite eyebrow. Excellent color and tissue match and transfer of hair-bearing tissue to the eyebrow and beard areas were achieved with no other complications. Satisfactory aesthetic results were gained. PMID- 11932594 TI - Endoscopically assisted reconstruction of male and female poland syndrome. AB - Poland syndrome produces deformities of the breast and chest wall that can be highly disfiguring in both men and women. Incisions from traditional surgical approaches can be unsightly, especially if a muscle harvest is used as part of the reconstruction. Herein is described a case series in which minimally invasive techniques were used to reconstruct defects in male and female patients with Poland syndrome. When evaluating patients with Poland syndrome, one should consider using minimally invasive techniques as an alternative reconstructive option. PMID- 11932595 TI - Circular excision of hemangioma and purse-string closure: the smallest possible scar. AB - Localized cutaneous infantile hemangioma acts like a tissue expander. This rapidly growing tumor can destroy elastic fibers or cause ulceration resulting in telangiectases, cutaneous laxity, scarring, and fibrofatty residuum. Although surgeons may dispute indications and timing, most would agree that the scar of resection should be minimized. For this reason, circular excision and purse string closure is particularly applicable for hemangioma at any stage of its evolution. The purposes of this study were to: (1) analyze the results of circular excision/purse-string closure in all three phases of the life cycle of hemangioma; (2) quantify dimensional changes after resection; and (3) compare the scars after theoretical single-stage lenticular excision with those after staged circular excision/purse-string closure. The authors retrospectively analyzed their experience in 25 children with localized hemangioma who underwent circular excision/purse-string closure from 1997 to 2000. Each hemangioma was measured preoperatively and the scars were measured at most recent follow-up (minimum, 6 months). Preoperative and postoperative dimensions were analyzed using SPSS statistical software. The study included 22 girls and three boys, with an average time to follow-up evaluation of 13.1 months. Twenty-one lesions were in the face and scalp, and five were in the extremity. Five tumors were resected in the proliferative phase (either because of ulceration, bleeding, or visual complications) and 21 were excised in the involuting or involuted phase. Six patients had a second-stage procedure: three had another circular excision and three had later lenticular excision. After single circular excision/purse-string closure, the mean long-axial diameter (length) decreased by 45 percent, the mean short-axial width (width) decreased by 73 percent, and the mean scar area was only 15 percent of the original area. All these differences were statistically significant (p = 0.001). The average width/length ratio decreased by 50 percent, indicating a tendency for scars to linearize. There was no difference in linearization for the three phases of hemangioma (p > 0.05); extremity scars became more linear that those on the face (p = 0.01). The authors devised a formula for scar length after lenticular excision/linear closure, assuming a conventional excisional ratio of 3:1 for a circular lesion. Using this equation, the authors predicted that mean scar length after circular excision, followed by lenticular excision, would be 72 percent shorter than the calculated scar that would result from conventional lenticular excision. In three patients who underwent this two-stage approach, the resultant scar was 69 percent shorter. Circular excision of hemangioma and purse-string closure reduces both the longitudinal and transverse dimensions and converts a large circular lesion into a small ellipsoid scar. If subsequent revision to a linear scar is desirable, its length will be the same or slightly less than the diameter of the original lesion. No other excision and closure technique results in a smaller scar. Another advantage of this method is minimal distortion of surrounding structures. PMID- 11932597 TI - The effectiveness of surgical and nonsurgical interventions in relieving the symptoms of macromastia. AB - In this report, the authors evaluate the effectiveness of breast reduction in alleviating the symptoms of macromastia by comparing baseline and postoperative health status using a series of well-validated self-report instruments. The study had a prospective design with a surgical intervention group and two control groups: a hypertrophy control group with bra cup sizes D or larger and a normal control group with bra cup sizes less than D. The effectiveness of nonsurgical interventions in relieving the symptoms of macromastia was also evaluated, both in the operative subjects and in the control groups. Surgical candidates and controls completed a self-administered baseline survey that consisted of the following validated and standardized instruments commonly used to evaluate outcomes: SF-36, EuroQol, Multidimensional Body-Self Relations Questionnaire (MBSRQ), and the McGill Pain Questionnaire (MPQ). A specially designed and validated instrument, the Breast-Related Symptoms (BRS), was also used. There were also questions about prior nonsurgical treatments, comorbid conditions, bra size, and a physical assessment. Additional information obtained on the operative subjects included surgical procedure data, resection weight, and complications. Approximately 6 to 9 months postoperatively, surgical subjects completed the same questionnaire as described above, and a final physical assessment was performed. The cohort included 179 operative subjects with matched preoperative and postoperative data sets, 96 normal controls and 88 hypertrophy controls. The women were predominantly Caucasian, middle-aged, well educated, and employed. Fifty percent of the operative subjects reported breast-related pain all or most of the time in the upper back, shoulders, neck, and lower back preoperatively compared with less than 10 percent postoperatively. Operative subjects and hypertrophy controls tried a number of conservative treatments, including weight loss, but none provided adequate permanent relief. Compared with population norms, the preoperative subjects had significantly lower scores (p < 0.05) in all eight health domains of the SF-36, and in the mental and physical component summary scores. After surgery, the operative subjects had higher means (better health) than national norms in seven of the eight domains and improved significantly from presurgical means in all eight domains (p < 0.05). Before surgery, the operative subjects reported high levels of pain with a Pain Rating Index (PRI) score from the MPQ of 26.6. After surgery, pain was significantly lower with a mean PRI score of 11.7, similar to that of our controls (mean PRI score, 11.2). Regression analysis was used to control for covariate effects on the main study outcomes. Among the operative subjects, benefits from breast reduction were not associated with body weight, bra cup size, or weight of resection, with essentially all patients benefiting from surgery.Breast hypertrophy has a significant impact on women's health status and quality of life as measured by validated and widely used self-report instruments including the SF 36, MPQ, and EuroQol. Pain is a significant symptom in this disease, and both pain and overall health status are markedly improved by breast reduction. In this population, conservative measures such as weight loss, physical therapy, special brassieres, and medications did not provide effective permanent relief of symptoms. PMID- 11932599 TI - Carpal tunnel release in the United States and Sweden: reimbursement patterns, cost for treatment, and return to work. AB - Cost containment plays an increasingly important role in health care, affecting providers and recipients. The present investigation addressed a limited number of factors that relate to the total cost for surgical treatment of carpal tunnel syndrome. The purpose of this study was to compare socioeconomic factors in two different societies and how worker's compensation plays a role in the cost, rehabilitation, and operative practices. To that purpose, the authors studied hospital and insurance records from a total of 123 female patients treated in two tertiary referral centers (University of Pittsburgh, Pa., and University of Umea, Sweden). The 123 patients were referred to three subgroups. Group A comprised patients from the University of Pittsburgh with worker's compensation (n = 34), group B comprised patients from the University of Pittsburgh without worker's compensation (n = 47), and group C comprised patients from the University of Umea (n = 42). The analyzed data showed a tendency toward longer duration of postoperative sick leave for those with worker's compensation than those without worker's compensation for patients from the University of Pittsburgh. However, operating room times and operating times, operative cost, use of postoperative therapy, and duration for sick leave were substantially less for patients treated at the University of Umea. There was no statistically significant difference in time off work between group A and group B. The results may indicate that the effect of compensation on a patient's ability or willingness to recover after treatment for carpal tunnel syndrome may be less important than factors that do not primarily relate to the patient or the surgical procedure. PMID- 11932598 TI - Soft-tissue flap coverage maximizes limb salvage after allograft bone extremity reconstruction. AB - Limb salvage after extremity tumor ablation may include the use of allograft bone. The primary complication of this method is infection of the allograft, which can lead to limb loss in up to 50 percent of cases. The purpose of this study is to evaluate the efficacy of primary muscle flap coverage in the setting of allograft bone limb salvage surgery. This study is a prospective review of all patients with flap coverage of extremity allografts over the 10-year period 1991 to 2001. There were 20 patients (11 male and nine female patients) with an average age of 28 years (range, 6 to 72 years). Flap coverage was primary in 16 patients and delayed in four. Delayed coverage was performed for failed wounds that did not have a primary soft-tissue flap. Pathologic findings included osteosarcoma in nine patients, Ewing sarcoma in five patients, malignant fibrohistiocytoma in two patients, chondrosarcoma in two patients, synovial sarcoma in one patient, and leiomyosarcoma in one patient. Allograft reconstruction was performed for the upper extremity in 12 patients and for the lower extremity in eight patients. Flap reconstruction was accomplished with 20 pedicle flaps in 17 patients (latissimus dorsi, 12; gastrocnemius, four; soleus, three; and fasciocutaneous flap, one) and four free flaps (rectus abdominis, three; latissimus dorsi, one) in four patients. All pedicled flaps survived. There was one flap failure in the entire series, which was a free rectus abdominis flap. This case resulted in the only limb loss noted. The follow-up period ranged from 1 to 50 months (average, 12.35 months). At the time of final follow-up, three patients were dead of disease and 17 were alive with intact extremities. The overall limb salvage rate in the setting of bone allograft and soft-tissue flap coverage was 95 percent (19 of 20). Reoperation for bone-related complications was required in 50 percent (two of four) of cases receiving delayed flap coverage compared with 19 percent (three of 16) of patients with primary flap coverage (statistically not significant). The results of this study support the use of soft-tissue flap coverage for allograft limb reconstruction. In this series, no limb was lost in the setting of a viable flap. Reoperation was markedly reduced in the setting of primary flap coverage. Pedicled or microvascular transfer of well-vascularized muscle can be used to wrap the allograft and minimize devastating wound complications potentially leading to loss of allograft and limb. PMID- 11932601 TI - Treatment of lateral ray polydactyly of the foot: focusing on the selection of the toe to be excised. AB - The purposes of this study were to evaluate the results of the operative treatment of lateral ray polydactyly and to consider appropriate surgical procedures, especially focusing on the selection of the toe, lateral toe or medial toe, to be resected. Twenty-two patients with lateral ray polydactyly foot (25 individual feet) at an average of 71 months' follow-up were included in this study. Cases were classified morphologically into three types on the basis of Hirase's configuration. In addition, these types were divided into two subtypes, metatarsal and phalangeal, on the basis of radiographic evaluation of the level of duplication. The clinical evaluations of the reconstructed toe were performed, and these results were investigated according to their morphologic classification and excised toe group. The distinctive problem of medial toe excision is valgus deformity. Eight of 25 toes retained persistent valgus deformity, and all of these cases were in the medial toe excision group. On the other hand, a distinctive problem in lateral toe excision is postoperative pain. Two patients suffered from postoperative pain in phalangeal type cases in the lateral toe excision group, and the remaining medial toe had a medial protuberant middle phalanx. The pain occurred at that protuberant point. Based on their experiences, the authors created an algorithm for selection of the toe to be excised. In metatarsal type cases, from a functional perspective, the toe that has a radiographically dominant metatarsus should be retained. On the other hand, in phalangeal type cases, the authors give priority to shape rather than function, and they excise the morphologically smaller toe independent of the condition of the phalanx as viewed on radiography. If the medial toe and the lateral toe are approximately the same size, the authors excise the lateral toe to avoid valgus deformity. When the lateral toe has severe valgus deformity that seems unlikely to be correctable intraoperatively, the lateral toe should be considered for excision even if it is larger than the medial toe. PMID- 11932602 TI - Vaginal reconstruction after extended radical pelvic surgery for cancer: comparison of two techniques. AB - An immediate partial or total vaginal reconstruction is frequently proposed in cases of exenterative or extended radical pelvic surgery for cancer treatment. One of the main complications after this reconstruction is the vagina obliteration caused by the healing process. This study compares the results of two different reconstructive techniques, particularly focusing on general complications and the risk of vaginal occlusion. A transversus rectus abdominis musculoperitoneal (TRAMP) composite flap has been performed in five cases, and an inverted inferior transverse rectus abdominis musculocutaneous flap (TRAM) has been used in another five cases. Recovery was uneventful in eight cases. One patient (case 5) developed an aortofemoral embolism requiring a bilateral transfemoral embolectomy and heparin administration. Another patient (case 9) experienced severe peritonitis because of the partial leak of the rectal anastomosis, and therefore a Mikulicz's colostomy was performed. Four patients who underwent the TRAMP flap developed a complete closure of the neovagina. In one patient with a TRAMP flap, a severe shortening (2 cm) of the neovagina occurred. Five patients out of five who underwent a reconstruction with a TRAM flap had a stable length of the neovagina (6 to 12 cm) and no shrinkage in diameter occurred, even though a vaginal stent was not used. The conventional inferior TRAM flap with a skin paddle seems to better maintain a stable length of the neovagina than the TRAMP composite flap with peritoneum. PMID- 11932604 TI - Immediate versus delayed midface distraction in a primate model using a new intraoral internal device. AB - The theoretic advantage of distraction osteogenesis of the craniofacial skeleton, especially in cases of severe midface retrusion and in the presence of maxillary scarring, is prevention of relapse following significant advancements. The purpose of this study is to demonstrate the utility of a new low-profile, intraoral, internal device for midface distraction at the conventional or high Le Fort I level. In addition, the present study compares the efficacy of immediate versus delayed distraction on subsequent maxillary relapse. Four adult rhesus Macaca mulatta monkeys were divided into two groups. Group 1 underwent immediate midface distraction; group 2 underwent delayed distraction. All four monkeys underwent a modified Le Fort I osteotomy through an upper buccal sulcus incision and bilateral application of the intraoral midface distraction devices. No other osteotomies or incisions were necessary. Immediate distraction, performed in group 1, entailed intraoperative activation of the devices and distraction of 10 mm followed by a 5-day lag period before postoperative activation and distraction of an additional 10 mm at the rate of 1 mm/day. Delayed distraction, performed in group 2, entailed a 5-day postoperative lag period before device activation and distraction of 20 mm at the rate of 1 mm/day. Both groups thus underwent 20 mm of midface distraction. All devices were removed 6 weeks after completion of distraction. All monkeys tolerated the devices and daily distraction uneventfully. On the basis of serial cephalograms and dental models obtained throughout the experimental period, there was no evidence of relapse in either the immediate or delayed groups 6 months after distraction. In addition, on the basis of histologic, ultrastructural, and dry skull analysis, no significant differences were observed in the quality of regenerate bone obtained when comparing the immediate and delayed distraction groups. Significant midface advancement is thus feasible using this new internal, intraoral distraction device, which presents several advantages over other internal devices that require coronal incisions and additional osteotomies to achieve midface advancement. In addition, immediate distraction may abbreviate the distraction period without adverse sequelae. PMID- 11932605 TI - Characterization of midface maxillary membranous bone formation during distraction osteogenesis. AB - The purpose of the study was to follow the early events in bone formation and neovascularization during maxillary distraction and after the consolidation period and to define the characterization of the new bone in the distracted area. Maxillary osteotomy was performed in seven sheep. In five animals, an external distraction device was used for maxillary lengthening of 20 mm at a rate of 1 mm/day for 20 days. Another two animals served as controls without distraction. Sequential biopsies were performed. The methods used for analysis were histologic, immunohistochemical, and ultrastructural by transmission electron microscopy. During the 5 days of latency, a fibrin clot was formed that after 5 days of distraction was replaced by granulation tissue, proliferating mesenchyme like cells, and capillaries. After 10 days of distraction, the regenerated tissue could be divided into three main zones and two transitional areas: a central zone occupied by many polygonal mesenchyme-like cells and spindle-shaped cells that proliferated intensively; two paracentral zones on both sides of the central zone in which many cells showed morphologic signs of apoptosis leading to a decreased number of fibroblast-like cells embedded in wavy collagen fibers; a transitional area from the central to the paracentral zone in which concentric cellular colonies were believed to represent a novel form of vasculogenesis; distal proximal zones, located on both sides of the paracentral zones and in continuation with the old bone, showed delicate new woven bone trabeculae that grew continuously in the direction of lengthening and gradually became mineralized; and a transitional area from the paracentral to the distal-proximal zones in which there was recruitment of preosteoblasts from the distracted tissue to the trabecular tips. These further differentiated into osteoblasts that contributed to the trabecular growth. The histologic feature pattern was similar after 15 and 20 days of continuous distraction. At the end of lengthening, after 20 days, delicate longitudinally oriented trabeculae continued to grow by recruiting preosteogenic cells from the central distracted tissue, became mineralized, and were rimmed by osteoblasts. After 6 weeks of retention, the trabeculae thickened and consisted of a mixture of lamellar and woven bone. In conclusion, the distraction force creates a pool of undifferentiated mesenchyme like cells with osteogenic potential and triggers capillary formation, a clear zonation can be observed during active lengthening, and new bone trabeculae begin to form between 5 and 10 days after distraction, soon become aligned with osteoblasts, and continue to grow as long as distraction force is applied. This characterization may help in any exogenous involvement with growth factors to improve bone quality. PMID- 11932606 TI - Callus stimulation in distraction osteogenesis. AB - Distraction osteogenesis has been described as in vivo tissue engineering. The ability to stimulate this process for the repair of bony defects or lengthening of congenitally shortened facial structures is likely to significantly impact the field of craniofacial surgery. The purpose of this study was to determine whether mechanical stimulation of the distracted rabbit mandible would accelerate the maturation of the bony callus when applied during the early consolidation period. Twenty adult New Zealand White rabbits underwent unilateral mandibular osteotomy. A uni-directional internal distractor device (Synthes, Paoli, Pa.) was positioned along a plane perpendicular to the line of osteotomy. After a 7-day latency period, distraction was commenced at a rate of 1.0 mm/day for 12 days in all animals. In a control group of 10 rabbits, a consolidation period of 8 weeks was observed before they were killed. In the experimental group of 10 rabbits, daily alternate compression and distraction of 1 mm (sequential compression and distraction) was performed for 3 weeks followed by a 5-week period of rigid fixation. Each animal received a dose of a fluorescent label at three different time points during the study: at the end of the distraction period, 3 weeks after the completion of the distraction phase, and 3 days before it was killed. All animals were killed 8 weeks after the completion of the distraction phase. Undecalcified histologic analysis and 3-point bending tests to failure were performed on the extracted mandibles. The results of the experimental and control groups were compared. Four animals in the control group and three animals in the experimental group were excluded from the study because of screw loosening resulting in distractor dislodgment or because of infection. On histologic analysis, cortical thickness at the center of the callus was found to be significantly greater in the experimental group compared with the control group when normalized to the contralateral hemimandible (83 percent versus 49 percent, respectively; p < 0.007). The ratio of cortical to cancellous bone in the distracted callus was uniformly found to be greater in the experimental specimens. The mineral apposition rate was calculated by using fluorescence microscopy and found to be significantly greater in the experimental group both during the period of sequential compression and distraction (3.2 microm/day versus 2.1 microm/day, p = 0.02) and after the period of sequential compression and distraction (1.4 microm/day versus 1.1 microm/day, p = 0.006). Mechanical testing revealed no significant differences in bending strength or stiffness between experimental or control groups (p = 0.54 and 0.47, respectively). This study has demonstrated that daily alternating compression and distraction of 1 mm amplitude during the early consolidation period has a stimulatory impact on callus formation with respect to osteoblastic activity, remodeling, and maturation of bone. Optimal timing and amplitude of sequential movement, long term biomechanical differences, and molecular pathways have yet to be elucidated. PMID- 11932607 TI - Reducing the vascular delay period in latissimus dorsi muscle flaps for use in cardiomyoplasty. AB - Although the mechanism by which vascular delay benefits skin flaps is not completely understood, this topic has been extensively studied and reported on in the literature. In contrast, little has been documented about the effects of vascular delay in skeletal muscle flaps. Recent animal studies tested the effectiveness of vascular delay to enhance latissimus dorsi muscle flap viability for use in cardiomyoplasty and found that it prevented distal flap necrosis. However, these studies did not define the optimal time period necessary to achieve this beneficial effect. The purpose of this study was to determine how many days of "delay" can elicit the beneficial effects of vascular delay on latissimus dorsi muscle flaps. To accomplish this, 90 latissimus dorsi muscles of 45 male Sprague-Dawley rats were randomly subjected to vascular delay on one side or a sham procedure on the other. After predetermined delay periods (0, 3, 7, 10, and 14 days) or a sham procedure, all latissimus dorsi muscles were elevated as single pedicled flaps based only on their thoracodorsal neurovascular pedicle. Latissimus dorsi muscle perfusion was measured using a Laser Doppler Perfusion Imager just before and immediately after flap elevation. The muscles were then returned to their original vascular beds, isolated from adjacent tissue with Silastic film, sutured into place to maintain their original size and shape, and left there for 5 days. After 5 days, the latissimus dorsi muscle flaps were dissected free, scanned again (Laser Doppler Perfusion Imager-perfusion measurements), and the area of distal necrosis was measured using digitized planimetry of magnified images. The authors' results showed that delay periods of 3, 7, 10, and 14 days significantly increased (p < 0.05) blood perfusion and decreased (p < 0.05) distal flap necrosis when compared with sham controls. On the basis of these findings, the authors conclude that in their rat latissimus dorsi muscle flap model the beneficial effects of vascular delay are present as early as 3 days. If these findings also hold true in humans, they could be useful in cardiomyoplasty by allowing surgeons to shorten the amount of time between the vascular delay procedure and the cardiomyoplasty procedure in these very sick patients. PMID- 11932609 TI - The behavior of fat grafts in recipient areas with enhanced vascularity. AB - Fat grafts are used for soft-tissue augmentation of various anatomic regions, most frequently for the improvement of facial contours. Resorption of the graft is the main problem, and several different procedures have been described to minimize this phenomenon. Using 25 New Zealand rabbits, the behavior of fat grafts in a highly vascularized recipient site was studied. The recipient sites prepared on the backs of the rabbits were divided into four regions. A capsule formation with silicone sheet application was accomplished in two of these recipient areas before the transplantation of the fat grafts. Fat grafts were placed in the other two recipient areas without any prior preparation. We prepared two types of fat tissue; in one the lobular structure was preserved and in the other it was manually crushed and rinsed with lactated Ringer's solution. The fat tissues with preserved lobular structure were placed in area I and area III. Manually crushed and rinsed fat tissues were placed in area II and area IV. In areas III and IV, a capsule formation with silicone sheet had been accomplished 3 weeks before grafting. Biopsy samples were obtained from these sites at the end of the first, third, sixth, and tenth months. Our aim was to observe the histologic fate of fat tissue in different recipient areas. The macroscopic and microscopic evaluation of the fat grafts in areas with silicone sheet indicated significant differences in the resorption time of the fat grafts; however, it was concluded that the significant resorption of the transplanted autologous fat tissue grafts at the end of the first year was an inevitable consequence of fat grafting. PMID- 11932608 TI - Presence and activity of nitric oxide synthase isoforms in ischemia-reperfusion injured flaps. AB - Nitric oxide is produced from the amino acid L-arginine by nitric oxide synthase, which has three known isoforms: (1) endothelial nitric oxide synthase and (2) brain nitric oxide synthase, both of which are constitutive nitric oxide synthase; and (3) inducible nitric oxide synthase. The authors' hypothesis is that after reperfusion injury, endothelial cell dysfunction leads to disruption of nitric oxide synthase-mediated nitric oxide production and that this may in part explain the deleterious effects of ischemia-reperfusion injury on tissue survival and blood reflow in flaps. An experiment was designed to study the effects of ischemia-reperfusion injury on the bioactivity of all three isoforms of nitric oxide synthase. Buttock skin flaps and latissimus dorsi myocutaneous flaps were elevated in eight pigs. Flaps on one side of the animal were randomized to receive 6 hours of arterial ischemia, whereas flaps on the other side served as controls. At 6 hours of ischemia and at 1, 4, and 18 hours after reflow, tissue biopsy specimens were obtained and were processed for both constitutive nitric oxide synthase and inducible nitric oxide synthase enzyme activity on the basis of the L-citrulline assay. In addition, specimens were processed for Western blot analysis of the three isoforms. The authors' results revealed three key findings: first, there was a statistically significant (p < 0.001) decrease in constitutive nitric oxide synthase activity of ischemia reperfusion-injured flaps as compared with controls in both skin and muscle for all time intervals measured. Second, Western blot analyses of endothelial nitric oxide synthase and brain nitric oxide synthase showed a significant decrease in the signal intensity in ischemic and reperfused tissue as compared with controls. Third, the inducible nitric oxide synthase isoform's activity and protein remained undetectable in both tissue types for all time points measured. The authors' data demonstrated that following ischemia-reperfusion injury in the pig flap model there was a disruption of constitutive nitric oxide synthase expression and activity, which may lead to decreased nitric oxide production. The significant decrease in nitric oxide synthase activity found in the current study may partly explain the mechanism of tissue damage in flaps subjected to ischemia reperfusion injury. Knowledge of the kinetics of nitric oxide synthase activity under conditions of ischemia-reperfusion injury has important implications for the choice and timing of delivery of therapeutic agents whose goal is to increase the bioavailability of nitric oxide in reperfused tissue. PMID- 11932611 TI - Vaginal delivery of monozygotic twins after bilateral pedicle TRAM breast reconstruction. PMID- 11932612 TI - Dorsal and laterothoracic radiodermatitis after interventional cardiography. PMID- 11932613 TI - Use of a web site in a multicenter plastic surgery trial: a new option for data acquisition. PMID- 11932615 TI - Matrix-periosteal flaps for reconstruction of nail deformity. PMID- 11932616 TI - Management and prevention of ongoing peristaltic contractions of the neovagina following rectosigmoid neocolpopoiesis in male transsexuals. PMID- 11932617 TI - Correction of secondary cleft lip deformities. AB - LEARNING OBJECTIVES: After studying this article, the practitioner should be able to (1) describe the common secondary deformities of the cleft lip, (2) determine the appropriate timing for surgical intervention to correct the deformities, and (3) determine the best method of addressing each of the individual secondary deformities of the cleft lip. Secondary deformities are common in children born with a cleft lip and palate. Patients with cleft lip deformity will undergo multiple surgical procedures early in life, so it is imperative to prioritize treatment of their secondary deformities and minimize the number of interventions needed. Of the many approaches used to correct these problems, surprisingly few work well consistently. As with all plastic surgery, the timing and procedure should be predicated on the severity of the deformity. PMID- 11932618 TI - Taking charge and doing the right thing. PMID- 11932619 TI - Antibiotic use in aesthetic surgery: a national survey and literature review. AB - Although much has been written regarding the use of antibiotics in surgery, no clear guidelines exist in the literature regarding the use of antibiotics in aesthetic surgery. To determine the pattern of antibiotic use in aesthetic surgery, a comprehensive survey was mailed to 6000 American Society of Plastic and Reconstructive Surgeons members and candidates. A total of 1767 completed responses were returned, which represents a response rate of 30 percent. The type, route, and duration of antibiotic administration are reported for 10 common cosmetic surgical procedures. The results of the survey confirmed that there is widespread use of antibiotics in aesthetic surgery. To provide guidelines for proper antibiotic use, the current literature was extensively reviewed. The authors found little or no correlation between reported clinical practice and the literature. It is hoped that this study will stimulate further investigation into this area of aesthetic surgery. PMID- 11932621 TI - The lumbrical fat graft: a replacement for lost upper eyelid fat. AB - Human eyelids seem to age earlier and more rapidly than do other regions of the face, making blepharoplasty (either age or familially indicated) an operation often performed earlier than other remedial procedures for facial aging. The traditional and more doctrinaire approach to upper blepharoplasty in which fat and orbicularis muscle are excised has led to a type of iatrogenia that often leaves the upper (and lower) eyelids hollowed and vacant, before their time. The notion of upper blepharoplasty as a simple operation is, therefore, to be questioned. The idea of normal and obligatory asymmetry as a key to recognizability is brought into relief when contemplating blepharoplasty. In questioning the judgments by which upper palpebral fat (and orbicularis muscle) is excised, the author suggests both a remedy-the lumbrical fat graft-and the need for a heightened awareness of the synergy such excisions make with the age related palpebral fat atrophies. This is a report of 35 cases (70 eyelids), over a 24-month period, during which the current technique was used. The technique is relatively simple, the anatomy well known, the learning slope shallow, the results gratifying, and the complications and problems few and benign. PMID- 11932623 TI - The "levator septi nasi muscle" and its clinical significance. AB - It is strange that all textbooks of anatomy describe the depressor septi nasi muscle singly, without an antagonist. Incidentally, in 1986, a small rod of soft tissue was found between the medial crura of the two alar cartilages during a rhinoplastic operation with the external approach technique of Anderson and Ries. From 1990 through 1995, anatomic dissections of the nasolabial region under 3.5x loupe magnification were performed on 14 Chinese formalin-preserved cadavers, one fresh Chinese cadaver, and one fresh American white female cadaver. The small soft-tissue rod was found in every one of the dissected cadavers, and it was seen to be a pair of muscles. Each one of these paired small muscles arose from the aponeurosis on the dorsum of the nose and inserted into the muscular substance of the upper lip at the base of the columella and to the anterior spine of the maxilla. Histologic examinations of these muscles stained with hematoxylin and eosin and Masson trichrome showed that they were striated muscles. According to its origin and insertion, this newly found muscle was called the "levator septi nasi." Its clinical significance in cleft lip deformity and its relations to the orbicularis oris muscle, the dermocartilaginous ligament of Pitanguy, and the nasal superficial musculoaponeurotic system of Letourneau and Daniel are all discussed. PMID- 11932625 TI - The management of anterior chest wall deformity in patients presenting for breast augmentation. AB - Anterior chest wall asymmetry is sometimes encountered in patients presenting for consideration of breast augmentation. The chest wall asymmetry or deficiency may be significant enough to consider reconstruction at the same time as breast augmentation in a small number of cases. Customized and prefabricated chest wall implants have been used in a variety of conditions including Poland syndrome, pectus excavatum, and sunken anterior chest. Careful moulage preparation and on table implant modification are needed to "seat" these implants on the skeletal chest wall under the pectoralis major muscle. The chest wall implant provides a base for the subsequent breast prostheses and fills up a bony deficit that cannot be camouflaged by the breast prostheses alone. PMID- 11932626 TI - Breast implant-related silicone granulomas: the literature and the litigation. AB - Formation of a granuloma is a common tissue response to the presence of a variety of foreign materials. A silicone granuloma is, by definition, a type of tissue reaction elicited occasionally by silicone. Granulomas of this sort have not engendered a great deal of attention in the scientific literature since their first description in 1964, and they were considered by most surgeons through the 1980s to be an incidental finding of little or no consequence. Since it is tangible and readily visible to juries, the silicone granuloma became a frequent issue in breast implant litigation in the 1990s. Although most other complaints in this litigation have been subjective or causally unrelated to silicone, the presence of a granuloma has been used as a basis of local complication claims and, more creatively, as a mechanism for hypothetical systemic disease occurrence. This review examines the basic science and the clinical literature pertaining to breast implant-related silicone granulomas, viewing them in a context of current clinical thought and frequent issues of litigation. Clinically apparent silicone granulomas are a relatively rare complication of breast implant placement and surgical resection is indicated when they are symptomatic or of diagnostic concern. There is no evidence in the peer-reviewed scientific literature to support frequent plaintiff theories in litigation that silicone granulomas play some role in implant-related systemic disease. In fact, the very existence of such diseases, themselves, is equally unsubstantiated. PMID- 11932628 TI - Application of an objective grading system for the evaluation of cosmetic surgical results. PMID- 11932629 TI - Ask and ye may or may not receive. PMID- 11932632 TI - Photographic copy identification of surgical instruments. PMID- 11932631 TI - A simple seroma aspirator. PMID- 11932634 TI - "Syringe in syringe" nipple splint. PMID- 11932635 TI - Outside the box. PMID- 11932636 TI - Should aspirin be discontinued 2 weeks before elective surgery? PMID- 11932637 TI - Treament of poststernotomy wounds with negative pressure. PMID- 11932638 TI - Radiation therapy does not cause skin ischemia. PMID- 11932639 TI - Intramasseteric hemangioma. PMID- 11932640 TI - Residency training in plastic surgery. PMID- 11932641 TI - Aesthetics in the third world. PMID- 11932642 TI - Congenital isolated absence of fifth metacarpal bone. PMID- 11932643 TI - The sagittohorizontal mandibular ramus osteotomy revisited: to avoid injury to the inferior alveolar nerve. PMID- 11932644 TI - Decompression of the common peroneal nerve: experience with 20 consecutive cases. PMID- 11932645 TI - Late reconstruction of zygomatic complex deformity with hydroxyapatite cement paste. PMID- 11932646 TI - Presutured purse-string suture and second purse-string suture for both reduction and closure of skin defects. PMID- 11932647 TI - Stamp and paper method: a superior technique for the walking track analysis. PMID- 11932648 TI - A practical method to cover the genital area in medical photography. PMID- 11932650 TI - A study of delayed diagnosis of facial nonmelanoma skin cancer. PMID- 11932651 TI - A simple tissue extensor. PMID- 11932652 TI - The role of serendipity in the evolving indications for integra dermal regeneration template. PMID- 11932653 TI - The suprafascial course of the direct paraumbilical perforator vessels. PMID- 11932654 TI - Regarding "Abdominal compartment syndrome after mesenteric revascularization". PMID- 11932655 TI - Cerebrospinal fluid drainage reduces paraplegia after thoracoabdominal aortic aneurysm repair: results of a randomized clinical trial. AB - OBJECTIVE: Despite the use of various strategies for the prevention of spinal cord ischemia, paraplegia and paraparesis continue to occur after thoracoabdominal aortic aneurysm (TAAA) repair. Although cerebrospinal fluid drainage (CSFD) is often used as an adjunct for spinal cord protection, its benefit remains unproven. The purpose of this randomized clinical trial was to evaluate the impact of CSFD on the incidence of spinal cord injury after extensive TAAA repair. METHODS: After randomization, 145 patients underwent extent I or II TAAA repairs with a consistent strategy of moderate heparinization, permissive mild hypothermia, left heart bypass, and reattachment of patent critical intercostal arteries. The repairs were performed with CSFD (n = 76) or without CSFD (n = 69). In the former group, CSFD was initiated during the operation and continued for 48 hours after surgery. The target CSF pressure was 10 mm Hg or less. RESULTS: The two groups had similar risk factors for paraplegia. Aortic clamp time, left heart bypass time, and number of reattached intercostal arteries were also similar in both groups. Thirty-day mortality rates were 5.3% (four patients) and 2.9% (two patients) for CSFD and control groups, respectively (P =.68). Nine patients (13.0%) in the control group had paraplegia or paraparesis develop. In contrast, only two patients in the CSFD group (2.6%) had deficits develop (P =.03). No patients with CSFD had immediate paraplegia. Overall, CSFD resulted in an 80% reduction in the relative risk of postoperative deficits. CONCLUSION: Perioperative CSFD reduces the rate of paraplegia after repair of extent I and II TAAAs. PMID- 11932656 TI - Functional outcome after thoracoabdominal aortic aneurysm repair. AB - OBJECTIVE: Repair of thoracoabdominal aortic aneurysms (TAAAs) is performed for the improvement of long-term survival and the preservation of function. The determination of functional outcome and the identification of predictors of survival and functional recovery after TAAA repair are key to proper patient selection. METHODS: This retrospective review of clinical data was performed in an academic medical center. The demographics, Crawford aneurysm type (I-18, II 33, III-22, IV-28), preoperative risk factors, operative characteristics, and postoperative complications and outcomes were recorded from the medical records for 101 consecutive patients who underwent TAAA repair (58 elective and 43 urgent/emergent). Functional status and living situation at hospital discharge and 12 months after discharge were determined from follow-up examination records or telephone contact with surviving patients. The patients then were categorized into "good" (survival, home, discharge to rehabilitation center, ambulatory) or "bad" (death, discharge to or residence in a long-term care facility, non ambulatory) outcomes. RESULTS: The postoperative mortality rate was 17.8% (10% in elective cases and 28% in urgent cases), and significant postoperative complications occurred in 77% of the cases (pulmonary complications in 41%, renal complications in 28%, and cord injury in 12%). The mean length of stay was 22.8 + 23.6 days, and at discharge, 80% of the patients were sent to home or rehabilitation and 20% were sent to long-term care facilities. At 1 year, 15 additional patients had died. All but two patients who had been initially discharged to rehabilitation had returned home, but only two patients who had been discharged to long-term care facilities had returned home and both were nonambulatory. Therefore, the survival rate at 1 year was 67%, and only 52.4% of the patients had a "good" outcome at 1 year (survival rate was 78% and rate of "good" outcome was 63% in patients who underwent elective TAAA repair). Independent predictors of postoperative death and "bad" outcome were age more than 75 years, preoperative heart disease, duration of visceral ischemia, use of left atrial femoral bypass graft, postoperative renal dysfunction, and number of organs failing after surgery. CONCLUSION: Survival and good functional outcome after TAAA repair is significantly less common than expected and is primarily predicted with intraoperative factors and postoperative complications. Improved operative techniques and limitation of visceral ischemia reperfusion injury may improve outcome after TAAA repair. PMID- 11932657 TI - Determinants of hospital length of stay after thoracoabdominal aortic aneurysm repair. AB - PURPOSE: Extended hospital length of stay (LOS) and consequent high costs are associated with thoracic and thoracoabdominal aortic aneurysm (TAAA) surgery. In this study, we examined factors that may influence LOS after TAAA repair. METHODS: Five hundred forty thoracic and TAAA repairs were performed by one surgeon between 1990 and 1999. The data were analyzed with multiple linear regression with appropriate logarithmic transformation. The predictor variables included patient demographics, disease extent, severity indicators, intraoperative factors, and postoperative complications. RESULTS: The median LOS was 15 days. Postoperative creatinine level of greater than 2.9 was the most important predictor of LOS, followed by spinal cord deficit, age, and pulmonary complication (all statistically significant with P <.05). A second model constrained to preoperative risk factors showed both age and complete diaphragmatic division to be associated with increased LOS. Preservation of the diaphragm led to reduced LOS by an average of 4 days. The adjunct cerebrospinal fluid drainage and distal aortic perfusion was associated with a decrease in LOS, although it did not reach statistical significance. CONCLUSION: Renal failure, spinal cord deficit, and pulmonary complication were the major determinants of LOS in patients for TAAA repair. This study shows that the preservation of diaphragmatic function and the use of the adjunct distal aortic perfusion and cerebrospinal fluid drainage may reduce hospital LOS. PMID- 11932658 TI - Minimally invasive vascular surgery for repair of infrarenal abdominal aortic aneurysm with iliac involvement. AB - PURPOSE: A minimally invasive vascular surgery (MIVS) technique for repair of infrarenal abdominal aortic aneurysm (AAA) with iliac involvement was evaluated, and its outcome was compared with conventional open repair. METHODS: Twenty patients with AAA with iliac involvement underwent treatment with bifurcated graft replacement with the MIVS technique. The procedure was performed via minilaparotomy, with the incision length determined according to the extent of the AAA obtained with ultrasound scanning and with the small intestine confined completely within the abdominal cavity. The proximal and distal operating fields were obtained with changing the patient position and arranging for the abdominal incision to be retracted cephalad and caudad. Perioperative courses in these 20 patients (the MIVS group) were analyzed in comparison with 14 patients who underwent conventional open repair, which was performed through the full midline laparotomy with the intestine simply covered with moistened towels (the conventional group). RESULTS: The MIVS technique for AAA repair was performed with a mean abdominal incision length of 8.4 cm and a range from 6.5 to 11.2 cm. The patients in the MIVS group showed earlier resumption of oral intake and ambulation in comparison with those patients in the conventional group (liquid diet: 1.1 +/- 0.3 days versus 2.9 +/- 1.4 days; P <.01; solid diet: 2.0 +/- 0.2 days versus 3.9 +/- 1.4 days; P <.01; ambulation: 2.1 +/- 0.8 days versus 4.3 +/- 2.3 days; P <.01), with comparable mortality and morbidity rates. Accordingly, the patients in the MIVS group were discharged earlier (20.7 +/- 6.3 days versus 33.9 +/- 12.6 days; P <.01), and total hospitalization charges were significantly decreased (2,232,791 +/- 200,747 Japanese yen versus 2,640,441 +/- 243,889 Japanese yen; P <.01). CONCLUSION: The MIVS technique allowed earlier postoperative recovery with comparable morbidity and mortality rates with the conventional technique and, therefore, saved hospital stay length and total hospitalization charges. Thus, the MIVS technique is considered as a new and effective minimally invasive technique for open AAA repair. PMID- 11932659 TI - Hemostatic markers before operation in patients with acutely symptomatic nonruptured and ruptured infrarenal abdominal aortic aneurysm. AB - BACKGROUND: In patients with acutely symptomatic but nonruptured abdominal aortic aneurysm (AAA), emergent repair is associated with an increased mortality rate as compared with semi-elective repair. Previous results have shown that ruptured but not asymptomatic AAA repair is associated with intense thrombin generation and inhibition of systemic fibrinolysis. The purpose of this study was to determine whether circulating markers of coagulation and fibrinolysis may be used to distinguish acutely symptomatic nonruptured and ruptured AAA. METHODS: A prospective study was performed of 44 patients who underwent emergency AAA repair for suspected rupture. Platelet count, fibrinogen level, prothrombin time, activated partial thromboplastin time, tissue plasminogen activator (t-PA) activity, plasminogen activator inhibitor (PAI) activity, prothrombin fragment (PF) 1+2 level, and D dimer level were measured before surgery. RESULTS: When compared with ruptured AAAs (n = 37), acutely symptomatic nonruptured AAAs (n = 7) were associated with increased fibrinogen level (P =.033), reduced activated partial thromboplastin time (P =.043), increased t-PA activity (P =.023), reduced PAI activity (P =.005), reduced PF 1+2 level (P =.001), and reduced D dimer level (P =.005; all P values determined with Mann-Whitney test). The differences in t PA activity (P =.01), PAI activity (P =.004), and PF 1+2 level (P =.01) persisted in patients whose conditions were normotensive. In all patients, a PF 1+2 level of greater than or equal to 2.5 nmol/L was associated with a sensitivity, specificity, and positive and negative predictive value for rupture of 89%, 86%, 97%, and 60%, respectively. In patients whose conditions were normotensive, PAI activity of greater than or equal to 16 AU/mL was associated with a sensitivity, specificity, and positive and negative predictive value of 83%, 100%, 100%, and 88%, respectively. CONCLUSION: These data show that acutely symptomatic nonruptured AAA is associated with increased systemic fibrinolysis (caused by reduced fibrinolytic inhibition) and reduced thrombin generation as compared with rupture. Preoperative hemostatic markers, particularly PF 1+2 level and PAI activity, may distinguish acutely symptomatic nonruptured from ruptured AAA. PMID- 11932660 TI - Expansion rates and outcomes for the 3.0-cm to the 3.9-cm infrarenal abdominal aortic aneurysm. AB - OBJECTIVE: This study was performed for the determination of the expansion rates and outcomes and for recommendations for the surveillance of the 3.0-cm to 3.9-cm abdominal aortic aneurysm (AAA). DESIGN: The study was observational with data from patients screened with ultrasound scanning for AAA at five Veterans Affairs Medical Centers for enrollment in the Aneurysm Detection and Management Trial. The eligibility requirements included: AAA from 3.0 cm to 3.9 cm in diameter and at least one repeat ultrasound scan more than 90 days after the initial screening. Patients also completed a questionnaire for demographic data and the determination of the presence of risk factors associated with AAA. The study endpoints included: 1, both mean and median expansion rates; 2, moderate expansion (>4 mm/year); 3, no expansion; 4, all causes of death; 5, AAA rupture; 6, expansion to 4 cm or more; 7, expansion to 5.0 cm or more; and 8, operative repair. RESULTS: Ultrasound scan screening results identified 1445 patients with 3.0-cm to 3.9-cm AAAs. Seven hundred ninety men met the ultrasound scan criterion of having at least two ultrasound scan studies during the study period, and these 790 men were used for this study. Mean AAA size was 3.3 cm, with an average follow-up period of 3.89 +/- 1.93 years. The median expansion rate was 0.11 cm/year. Expansion rates were significantly different (P <.001) between 3.0-cm and 3.4-cm cm AAA and 3.5-cm and 3.9-cm AAA. There were no reported AAA ruptures during the study period, although cause of death data were available in only 43% of the patients. Few 3.0-cm to 3.9-cm AAAs expanded to 5.0 cm or more during the study period. The patients with 3.0-cm to 3.9-cm AAAs who underwent operative repair during the study period were younger, had larger initial AAA diameters, and had more rapid expansion rates. CONCLUSION: AAAs of 3.0 cm to 3.9 cm expanded slowly, did not rupture, and rarely had operative repair or expanded to more than 5.0 cm in our study of male patients. Expansion rates and the incidence rate of operative repair are more common in the 3.5-cm to 3.9-cm AAA when compared with the 3.0-cm to 3.4-cm AAA. PMID- 11932661 TI - Pathogenetic heterogeneity of in-stent lesion formation in human peripheral arterial disease. AB - PURPOSE: Intimal hyperplasia is known to promote in-stent restenosis after vascular stent implantation. Although previous studies have presented a variety of evidence that suggests the mechanisms of intimal lesion formation, it is still controversial which factor(s) predominantly contribute to the development of in stent restenosis. In this study, we hypothesized that heterogeneous mechanisms coexist in the same lesion and then assessed the validity with resected whole arteries with in-stent lesions. METHODS: Whole arterial specimens with in-stent lesions were surgically resected from five patients who had undergone implantation of a Palmaz-Schatz stent 7 to 19 months previously and were histologically analyzed. For assessment of the pathogenetic heterogeneity of in stent lesions, we divided each cross section into three parts: the inner intima within 250 microm from the luminal surface (zone A), the area surrounding the stent struts within 250 microm from the strut hole (zone B), and the remaining part of the intimal layer (zone C). We then evaluated cell density, cell replication, and cellular composition in each zone. Cell replication and cellular composition were analyzed with immunohistologic staining with antibodies against proliferating cell nuclear antigen and cell-specific antibodies. Each section was also stained with Alcian blue or Elastica van Gieson method for detection of matrix components. RESULTS: In all samples, the cell density of zone A was significantly higher than that of zone B (P <.05). Proliferating cell nuclear antigen staining results showed significantly higher cell replication in zone A as compared with that in other zones (P <.05). To the contrary, cell-specific immunostaining results revealed marked accumulation of leukocytes, macrophages, and T lymphocytes in zone B (P <.05). Regarding matrix components, proteoglycan was predominantly stained around stent struts and in the inner intima. CONCLUSION: The data of this study showed that two different pathogenetic processes in different zones possibly contributed to in-stent lesion formation at the same time. One process was an increase of cell number in the inner intima, which was the result of a prolonged increase of cell replication. The other process was accumulation of matrix around stent struts, which was suggested to be linked to infiltration of inflammatory cells in the same zone. PMID- 11932662 TI - Potential impact of therapeutic warfarin treatment on type II endoleaks and sac shrinkage rates on midterm follow-up examination. AB - OBJECTIVE: Successful endovascular aortic aneurysm repair depends on exclusion and spontaneous thrombosis of the aneurysm sac. The need for chronic postoperative anticoagulation therapy could limit the applicability of this technology with delay or prevention of sac thrombosis resulting in endoleak formation and altered remodeling of the aneurysm sac. The purpose of this study was the determination of whether chronic therapeutic anticoagulation therapy with warfarin was associated with an increased incidence rate of early or delayed postoperative endoleaks or altered rates of reduction in aneurysm sac maximum diameter. METHODS: Two hundred thirty-two consecutive patients underwent abdominal aortic endografting during a 32-month period. The data were recorded prospectively with a current mean follow-up period of 18 months. The patients with endoleaks identified with 30-day postoperative computed tomographic scan angiograms subsequently underwent selective arteriography to characterize the source. The patients who underwent chronic warfarin therapy that resulted in a therapeutic internationalized normalized ratio comprised the study group. The control group was defined as all the patients with healthy coagulation profiles. RESULTS: Thirty-six patients (15%) were undergoing warfarin therapy after surgery, and their conditions were chronically maintained with a therapeutic international normalized ratio. Forty-three patients (18%) had endoleaks on 30 day computed tomographic scan angiographic results. There were 39 patients with type II endoleaks and four patients with type I endoleaks. None of the type I endoleaks occurred in patients who were undergoing warfarin therapy, and all endoleaks were repaired with either proximal or distal covered extensions. At 30 days, seven patients (19.4%) undergoing chronic warfarin therapy had type II endoleaks as compared with 36 controls (18.4%; P =.798). Four patients had delayed type II endoleaks develop, two in the control group and two in the warfarin group (P =.3). Ten control individuals (31%) had spontaneous resolution of type II endoleaks develop, whereas spontaneous endoleak thrombosis was not observed in the warfarin group (P =.33). Aneurysm sac remodeling assessed with mean percent reduction in maximum sac diameter at 12 months revealed a statistical difference between the control group (17.5%) and the warfarin group (7.6%; P =.04). CONCLUSION: Warfarin treatment is not associated with an increase in the incidence rate of early or delayed postoperative endoleaks. However, the rate of reduction in maximum aneurysm sac diameter after aortic endografting is slower in patients who undergo therapeutic warfarin therapy at 1-year follow-up examination, a statistically significant difference from the control group. In addition, type II endoleaks may be less likely to undergo spontaneous thrombosis in patients who undergo warfarin therapy. PMID- 11932663 TI - Endograft limb occlusion and stenosis after ANCURE endovascular abdominal aneurysm repair. AB - OBJECTIVE: The purpose of this study was to define the incidence and treatment of endograft limb stenosis or occlusion (endograft limb dysfunction [ELD]) in a single center with the ANCURE unsupported bifurcated or aortouniiliac endograft by using intraoperative completion angiography and postoperative color duplex ultrasound scanning (CDU). METHODS: Sixty-seven endografts (58 bifurcated, 9 uniiliac) were implanted between February 1996 and July 2000. Intraoperative completion aortography was performed in every patient. Postoperative assessment of the endograft consisted of CDU and computed tomography scanning and kidney, ureter and bladder radiographs within 7 days of implantation, at 3 and 6 months after the operation, and every 6 months thereafter. RESULTS: At the time of endograft implantation, widely patent normal-appearing endograft limbs were revealed by means of the initial completion angiogram in 58 of 67 patients (group 1). ELD subsequently developed in seven of these 58 patients (13.4%). The results of the completion angiogram were not normal in the remaining nine patients (group 2), leading to the deployment of a self-expanding stent within the endograft limbs. The results of subsequent angiography were normal. No ELD has occurred in any patient in group 2 to date. The primary assisted patency rate at 30 months was 88% +/- 5.2% for group 1 versus 100% +/- 0% for group 2 (P = not significant, Log-rank test). Postoperative ELD occurred in seven patients (10.4%). Endovascular graft thrombosis occurred in three patients (3 endograft limbs). In each case, an endovascular approach was attempted; however, the guidewire would not traverse the occluded endovascular graft limb. Revascularization was accomplished by means of femorofemoral bypass grafting. Endovascular graft stenosis occurred in four patients (4 endograft limbs). Three patients with bifurcated endografts and limb stenosis who had no symptoms diagnosed by means of CDU were successfully treated by means of balloon angioplasty with self-expanding stent implantation, and the endograft limbs remained patent at 3, 5, and 26 months follow-up. The remaining patient who had an aortouniiliac endograft with recurrent severe stenoses underwent endograft explantation and aortobifemoral bypass grafting. The overall incidence of ELD during or after endovascular abdominal aortic aneurysm repair was 23.8% (16 of 67 patients). CONCLUSION: Unsupported endografts are at risk for developing ELD. The use of stents for limb support at the time of the initial endograft implantation may prevent subsequent ELD and bears further study. Endograft limb occlusion usually presents with acute severe ischemic symptoms, and the failure of operative thrombectomy necessitates femorofemoral artery bypass grafting. Endograft limb stenosis is identified by means of CDU surveillance in the postoperative period. Prompt treatment with percutaneous transluminal angioplasty/stent yields satisfactory primary assisted patency. Intraoperative intravenous ultrasound scanning, oblique angiograms, pressure gradients, and completion angiography may be necessary to detect and treat ELD. PMID- 11932664 TI - Safety and efficacy of femoral-based hemodialysis access grafts. AB - OBJECTIVE: Arteriovenous fistulae traditionally have been placed in the upper extremity. Experience with groin hemodialysis access has been discouraging because of high infection rates and associated limb amputation. We reviewed our experience with angioaccess grafts in the groin to assess safety and viability in our hemodialysis patient population. METHODS: A retrospective review was performed of all groin hemodialysis access grafts placed at a single tertiary care center between June 1990 and February 1998. Demographics, complications, and subsequent treatment were recorded. Graft patency and infection rates were analyzed with life-table analysis. RESULTS: Data were collected on 73 graft insertions. A total of 52 episodes of thrombosis occurred in 26 grafts. Primary patency rate was 71% at 1 year. Secondary patency rate was 83% at 1 year. There was a 22% incidence rate of infection. CONCLUSION: We conclude that the incidence rate of infection and thrombosis in our series of femoral-based hemodialysis grafts is comparable with rates reported in the literature for upper extremity polytetrafluoroethylene angioaccess grafts. Although not considered a first choice, femoral artery-based hemodialysis access is a viable option when arteriovenous fistulae in the upper extremity cannot be constructed. PMID- 11932665 TI - Intravascular ultrasound scan evaluation of the obstructed vein. AB - PURPOSE: The purpose of this study was the comparison of intravascular ultrasound scanning (IVUS) with transfemoral venography in the assessment of chronic iliac vein obstruction. METHODS: IVUS and standard, single-plane, transfemoral venography were performed in 304 consecutive limbs during balloon dilation and stenting of an obstructed iliac venous segment. The appearance of the obstruction was described, and the degree of stenosis (maximal diameter reduction) was estimated with venography and IVUS. The stenotic area was derived with diameter calculations (pir(2)) and also was measured with the built-in software of the IVUS apparatus before and after dilation and stenting in 173 limbs. Preoperative hand/foot differential pressure and preoperative dorsal foot venous and intraoperative transfemoral hyperemia-induced pressure elevations after intra arterial injection of papaverine hydrochloride were measured. RESULTS: With IVUS, fine intraluminal and mural details were detected (eg, trabeculation, frozen valves, mural thickness, and outside compression) that were not seen with venography. The median stenosis (with diameter reduction) on venographic results was 50% (range, 0 to 100%) and on IVUS results was 80% (range, 25% to 100%). In a comparison with IVUS as the standard, venography had poor sensitivity (45%) and negative predictive value (49%) in the detection of a venous area stenosis of >70%. The actual stenotic area was more severe when measured directly with IVUS (0.31 cm(2); range, 0 to 1.68 cm(2)) versus derived (0.36 cm(2); range, 0 to 3.08 cm(2); P <.001), probably as a result of the noncircular lumen geometry of the stenosis. No correlation was found between any of the preoperative or intraoperative pressure measurements and degree of stenosis with or without collaterals. When collaterals were present, a more severe stenosis (median, 85%; range, 25% to 100%) was observed (versus a 70% stenosis in the absence of collaterals; range, 30% to 99%; P <.001), along with actual stenotic area (with collaterals: median, 0.24 cm(2); range, 0 to 1.18 cm(2); without collaterals: median, 0.45 cm(2); range, 0.02 to 1.68 cm(2); P <.01) and a higher rate of hyperemia-induced pressure gradient (> or =2 mm Hg; with collaterals, 34%; without collaterals, 11%; P <.05). CONCLUSION: Venous IVUS appears to be superior to single-plane venography for the morphologic diagnosis of iliac venous outflow obstruction and is an invaluable assistance in the accurate placement of venous stents after venoplasty. No preoperative or intraoperative pressure test appears to adequately measure the hemodynamic significance of the stenosis. In lieu of adequate hemodynamic tests, IVUS determination of morphologically significant stenosis appears to be presently the best available method for the diagnosis of clinically important chronic iliac vein obstruction. Collateral formation should perhaps be looked on as an indicator of a more severe stenosis, although significant obstruction may exist with no collateral formation. PMID- 11932666 TI - Inflammatory response in the acute phase of deep vein thrombosis. AB - OBJECTIVE: Deep vein thrombosis (DVT) is a multifactorial disease. Recently, inflammation has been suggested as a risk factor for DVT. The question is whether inflammation is a cause of venous thrombosis or rather a result of the thrombotic process. METHODS: We studied the inflammatory response in the acute phase of DVT with interleukin-6, interleukin-8, and C-reactive protein (CRP) as inflammatory markers. Plasma concentrations were measured on the day of admission (day 0) in 40 patients with acute DVT confirmed with phlebography and in 33 patients with clinical suspicion of DVT but negative phlebography results (controls). In patients with DVT, inflammatory markers were also examined on five subsequent days. RESULTS: On day 0, the median concentrations in plasma of interleukin-6, interleukin-8, and CRP were 15.0 pg/mL (range, <3 to 70 pg/mL), 7.0 pg/mL (range, <3 to 76 pg/mL), 37.5 mg/L (range, <7 to 164 mg/L), respectively, in the patient group and less than 3 pg/mL (range, <3 to 11 pg/mL; P <.001), 6.0 pg/mL (range, <3 to 52 pg/mL; P =.08), and 5.0 pg/L (range, <7 to 66 pg/L; P <.001), respectively, in the controls. During the next days, interleukin-6 concentration showed a gradual decline in patients with DVT from 15.0 to 5.5 pg/mL (P <.001), interleukin-8 concentration was relatively constant in time, and CRP concentration declined from 37.5 to 21.5 mg/L (P =.01). CONCLUSION: Our data show an apparent inflammatory response with highest measured concentrations of inflammatory markers on the day of admission and a subsequent decrease during the next days. This response supports the hypothesis that elevated inflammatory markers are a result rather than a cause of venous thrombosis. PMID- 11932667 TI - Early diagnosis of deep vein thrombosis in female patients who undergo total knee arthroplasty with measurement of P-selectin activation. AB - BACKGROUND: Deep vein thrombosis (DVT) remains a leading cause of postoperative morbidity and mortality in patients who undergo total knee arthroplasty (TKA). Although patients with previous thrombotic episodes are inherently at a higher risk for subsequent episodes of DVT, it remains difficult to predict such an occurrence and to make a diagnosis in early stages. One potentially useful assay that can be used in the determination of changes of coagulation among patients who undergo arthroplasty is platelet activation. The goal of this study was to establish a predictive value for DVT with measurement of P-selectin levels that could help in planning appropriate perioperative management strategies for patients at high risk for DVT. METHODS: A total of 52 patients who underwent TKA with general anesthesia underwent contrast venography on the 5th postoperative day. Platelet activation before and after operation was measured with platelet surface expression of P-selectin with flow cytometry in these two groups of patients for TKA. None of the patients underwent any anticoagulation therapy. RESULTS: Nineteen of the 52 patients for TKA showed radiologic evidence of DVT, whereas 33 patients for TKA had no radiologic signs of DVT. There was no difference in platelet activation at baseline, which was 1 hour before induction of anesthesia, between the two groups (P >.05) as measured with P-selectin assays. Differences were noted between the two groups on the 5th day after operation, wherein P-selectin was expressed in only 2.72% +/- 0.9% (mean +/- standard deviation) of platelets in patients for TKA with healthy venogram results. This differed significantly from platelets in patients for TKA with DVT, who had P-selectin expression of 6.56% +/- 3.1% (mean +/- standard deviation; P <.01). Sensitivity for the diagnosis of DVT with P-selectin assay was calculated to be 74%, and specificity was found to be 94%. CONCLUSION: The findings showed that radiologically confirmed DVT in patients for TKA surgery with general anesthesia is associated with an elevated number of activated platelets. Perioperative assessment of P-selectin may predict the early onset of DVT in patients who undergo high risk surgical procedures like TKA. This laboratory assay may help prevent the occurrence of the fatal events caused by DVT with use of early therapeutic intervention, such as heparinization. PMID- 11932668 TI - The mechanism of venous valve closure in normal physiologic conditions. AB - PURPOSE: The purpose of this investigation was to study the mechanism of venous valve closure in physiologic conditions in situ, particularly the clarification of the role of reversed flow through the valve in the closure of the valve cusps. A second purpose was the study of temporal relations between movements of the valve cusps, changes in geometry of venous sinus, and blood flow. METHODS: Twelve healthy volunteers underwent examination with duplex ultrasound scanning. The time relationship between the flow and venous valve movements was analyzed on the basis of real time ultrasound scan records of the saphenofemoral junction. The size and shape of the common femoral vein and the proximal greater saphenous vein were measured with a planimeter. RESULTS: Reverse flow below the valve was registered in only one of the 12 studied subjects. The maximum peak velocity of the reverse flow in this case was 0.8 cm/s and the duration was 0.2 seconds. The reverse flow was registered just before and just after valve closure. The mean duration of the outflow wave below the valve was 816 +/- 57 ms, which was not significantly different from the time of the valve cycle (819 +/- 55 ms; P =.864). The closure of the valve coincided with the decrease of the flow velocities. The first detectable movement of the valve cusps occurred 108 +/- 21 ms after the beginning of the flow deceleration. The mean time interval from the first cusp movement to the complete closure of the valve was 139 +/- 51 ms. Both the size and the shape of the sinus changed during the valve cycle. The size of the sinus increased as much as 127% of the baseline value (1.79 +/- 0.25 cm at baseline; 2.27 +/- 0.23 cm maximum; P =.02), and it became more spheric in shape. The changes in size and shape of the sinus coincided with the movements of the valve cusps. The first detectable change in the sinus size occurred 80 +/- 30 ms after the first detectable movement of the cusps toward closure. CONCLUSION: Reverse flow through the valve is not necessary for the closure of the venous valve. The closure of the venous valve coincides with the decrease of the flow velocities and the ballooning of the sinus (increase of size and the curvature of the valve sinus). PMID- 11932669 TI - The prevalence of thrombophilia in patients with chronic venous leg ulceration. AB - BACKGROUND: Thrombophilia is increasingly recognized as a risk factor for deep venous thrombosis (DVT), which in turn is a major risk factor for chronic venous ulceration (CVU). However, the relationship between thrombophilia and CVU remains unknown. The aim of this study was to define the prevalence of thrombophilia in patients with CVU and to determine whether this is associated with a history or duplex scan evidence of DVT. METHODS: Eighty-eight patients with CVU were prospectively studied. The patients underwent clinical assessment and duplex ultrasound scanning. Blood was drawn for antithrombin, proteins C and S, activated protein C resistance, factor V Leiden, prothrombin 20210A, lupus anticoagulant, and anticardiolipin antibodies. RESULTS: The study included 35 men with a median age of 61 years (interquartile range, 45 to 72 years) and 53 women with a median age of 76 years (interquartile range, 69 to 82 years). Thirty-six percent of the patients had either a history or duplex scan evidence suggestive of previous DVT. The following abnormalities were detected: four, five, and six cases of antithrombin, protein C, and protein S deficiencies, respectively; 14 cases of activated protein C resistance; 11 cases of factor V Leiden mutation; three cases of prothrombin 20210A mutation; eight cases of lupus anticoagulant; and 12 cases of anticardiolipin antibodies. Thrombophilia was not significantly related to previous DVT, deep reflux, or disease severity. CONCLUSION: Patients with CVU have a 41% prevalence rate of thrombophilia. This rate is two to 30 times higher than the rate of the general population but is similar to that reported for patients with previous DVT. However, in patients with CVU, thrombophilia does not appear to be related to a history of DVT, a pattern of reflux, or severity of disease. Many patients with CVU may have unsuspected postthrombotic disease. PMID- 11932670 TI - Recurrence of chronic venous ulcers on the basis of clinical, etiologic, anatomic, and pathophysiologic criteria and air plethysmography. AB - INTRODUCTION: Leg ulcers associated with chronic venous insufficiency (CVI) frequently recur after healing. The risk of recurrence has not been well defined for patients in different anatomic and hemodynamic groups. We reviewed the risk of ulcer recurrence on the basis of clinical, etiologic, anatomic, and pathophysiologic criteria and hemodynamic characteristics of the affected limb as assessed with air plethysmography (APG). METHODS: Ninety-nine limbs with class 6 CVI were assessed clinically and with standing duplex ultrasound scanning and APG for the definition of clinical, etiologic, anatomic, and pathophysiologic criteria. Leg ulcers were treated with high-pressure compression protocols. Surgical correction of venous abnormalities was offered to patients with appropriate conditions. After ulcer healing, the limbs were placed in compressive garments and followed at 6-month intervals for ulcer recurrence. RESULTS: The mean patient age was 54.3 years, and 46% of the patients were female. Corrective venous surgery was performed in 37 limbs. The mean follow-up time for all 99 limbs was 28 months. The ulcer recurrence rate with life table was 37% +/- 6% at 3 years and 48% +/- 10% at 5 years. The patients who underwent venous surgery had a significantly lower recurrence rate (27% +/- 9% at 48 months) than did those patients who had not undergone surgery (67% +/- 8% at 48 months; P =.005). The patients with deep venous insufficiency (DVI; n = 51) had significantly higher recurrence rates (66% +/- 8% at 48 months) than did the patients without DVI (n = 48; 29% +/- 9% at 48 months; P =.006). This difference was significant even after accounting for the effects of surgery (P =.03). The hazard ratio of ulcer recurrence increases by 14% for every unit increase in the venous filling index (VFI; P =.001). This remains significant even after accounting for the effects of surgery (P =.001). The combination of DVI and a VFI of more than 4 mL/s yields a risk of ulcer recurrence of 43% +/- 9% at 1 year and 60% +/- 10% at 2 years. CONCLUSION: Leg ulcers associated with CVI have a high rate of recurrence. Ulcer recurrence is significantly increased in patients with DVI and in patients who do not have venous abnormalities corrected surgically. The VFI obtained from APG is useful in the prediction of increased risk for recurrence, particularly in association with anatomic data. PMID- 11932671 TI - Endovenous treatment of the greater saphenous vein with a 940-nm diode laser: thrombotic occlusion after endoluminal thermal damage by laser-generated steam bubbles. AB - PURPOSE: Despite a rapid spread of the technique, very little is known about the laser-tissue interaction in endovenous laser treatment (EVLT). We evaluated EVLT of the incompetent greater saphenous vein (GSV) for efficacy, treatment-related adverse effects, and putative mechanisms of action. METHODS: Twenty-six patients with 31 limbs of clinical stages C(2-6), E(P), A(S,P), P(R) with incompetent GSV proven by means of duplex scanning were selected for EVLT in an outpatient setting. A 600-microm fiber was entered into the GSV via an 18-gauge needle below the knee and proceeded to the saphenofemoral junction (SFJ). After infiltration of tumescent local anesthesia, multiple laser pulses of 15 J energy and a wavelength of 940 nm were administered along the vein in a standardized fashion. D-dimers were determined in peripheral blood samples 30 minutes after completion of EVLT in 16 patients and on postoperative day 1 in 20 patients. One GSV that was surgically removed after EVLT was examined by means of histopathology. Additionally, an experimental in vitro set-up was constructed as a means of investigating the mechanism of laser action within a blood-filled tube. RESULTS: A median of 80 laser pulses (range, 22-116 laser pulses) were applied along the treated veins. On days 1, 7, and 28, all limbs except one (97%) showed a thrombotically occluded GSV. In one patient, the vessel showed incomplete occlusion. The distance of the proximal end of the thrombus to the SFJ was a median 1.1 cm (range, 0.2-5.9 cm) in the remaining patients. Adverse effects in all 26 patients were ecchymoses and palpable induration along the thrombotically occluded GSV that lasted for 2 to 3 weeks. In two limbs (6%), thrombophlebitis of a varicose tributary required oral treatment with diclofenac. D-dimers in peripheral blood were tested with normal results in 14 of 16 patients 30 minutes after completion of the procedure and elevated results in 7 of 20 patients at day 1 after EVLT. However, an increase of D-dimers from day 0 to day 1 was observed in 15 of the 16 patients undergoing tests 30 minutes after EVLT and on day 1. The 940-nm laser was demonstrated by means of in vitro experiments and the histopathological examination of one explanted GSV to act by means of indirect heat damage of the inner vein wall. CONCLUSION: EVLT of the GSV with a 940-nm diode laser is effective in inducing thrombotic vessel occlusion and is associated with only minor adverse effects. Laser-induced indirect local heat injury of the inner vein wall by steam bubbles originating from boiling blood is proposed as the pathophysiological mechanism of action of EVLT. PMID- 11932672 TI - Retrojugular approach for carotid endarterectomy: a prospective cohort study. AB - The purpose of this study was the examination of the feasibility of the retrojugular approach for carotid endarterectomy and the investigation of its safety and effectiveness. A consecutive prospective cohort of 43 patients who underwent carotid endarterectomy with the retrojugular approach was compared with a retrospective cohort of 43 patients who underwent surgery with the standard antejugular technique. Age, sex, comorbidity, diagnostic investigations, and indications for surgery were comparable in both groups. In the retrojugular group, a carotid shunt was used in 18 patients (42%) and a patch closure in 15 patients (35%), and in the antejugular group, a carotid shunt was used in 43 patients (100%) and a patch closure in 20 patients (47%). There were no perioperative deaths, strokes, nerve injuries, or 30-day postoperative neurologic events. Wound hematomas were equally distributed in both groups (5%). Transient hoarseness was present in two patients in the antejugular group. The mean operative time was 72 +/- 15 minutes for the retrojugular technique and 100 +/- 27 for the antejugular technique (P <.0001). The hospital stay was 2 +/- 1 days in both groups. In conclusion, the retrojugular approach appears to be safe and the operative time may be shorter than with the antejugular approach. PMID- 11932673 TI - Major carotid plaque surface irregularities correlate with neurologic symptoms. AB - PURPOSE: Many studies have linked carotid plaque surface irregularities with stroke risk, but this relationship has been obscured by the limited ability of available imaging modalities to resolve plaque surface morphology. To address this issue, we performed a prospective study correlating the presenting neurologic symptoms of patients with high-resolution magnetic resonance imaging (MRI; 200 microm) studies of ipsilateral plaque surface invaginations and ledges, lumen shape, and the location of the plaque bulk creating the stenosis. METHODS: One hundred patients, 17 women and 83 men, 45 to 81 years old (mean, 68 years) underwent surgery. Forty-five patients had a transient ischemic attack (TIA) or stroke as the indication for surgery, and 55 patients had no symptoms. Angiograms were obtained in 50 patients. Carotid plaques were removed "en bloc" and placed in gadolinium doped saline for imaging in a Siemens Symphony, 1.5T scanner with a custom-built transmit-receive radiofrequency coil. The resulting slice thickness was 200 microm, with 200 microm by 200 microm in plane resolution. The MRI data and angiograms were reviewed by using National Institutes of Health Image software and read by consensus. A surface irregularity was categorized as a ledge or ulcer and measured by using electronic calipers. Luminal shape was determined at the point of maximal stenosis with a "slice" set at 90 degrees to the lumen axis. The location of the maximal stenosis was recorded. In the internal carotid artery, plaque bulk was designated to be on the flow divider wall or non-flow divider wall. RESULTS: The mean maximal stenosis was 81.5% +/- 12.0%. Surface contour irregularities were found in 80 plaques. Thirty-five plaques were graded as having major surface contour irregularities, and 45 plaques were graded as having minor irregularities. There was a significant correlation between major surface irregularity and TIA or stroke (P <.01). Irregular plaques were identified with angiography, but the irregularity in size was underestimated (P <.01). Only 28% of plaques had circular lumens; 50% had elliptical lumens, and 22% had either crescentic or multi-lobular lumens. The maximal stenosis was located in the internal carotid artery in 82 plaques, the bifurcation in 17 plaques, and the common carotid artery in one plaque. CONCLUSION: Surface irregularities were revealed by means of submillimeter resolution of the carotid plaques with MRI to be common, but only the presence of major irregularities correlated with the patient having TIA or stroke. Lumen shape and plaque location did not appear to predict stroke risk, but may effect imaging accuracy in determining the degree of stenosis. These data further define the relationship of plaque irregularity and cerebrovascular symptoms caused by atheroemboli. PMID- 11932674 TI - Gender as a primary predictor of outcome after carotid endarterectomy. AB - OBJECTIVE: Post hoc analysis results of the North American Symptomatic Carotid Endarterectomy Trial and the Asymptomatic Carotid Atherosclerosis Study suggest that carotid endarterectomy (CEA) may not be as efficacious in women as it is in men. This study was undertaken for the evaluation of whether there is a difference between men and women in early postoperative outcome after CEA and whether such a difference is consistent across other predictors. METHODS: We conducted a retrospective review of all CEAs entered into our departmental registry between January 1, 1989, and November 30, 2000. A total of 3422 CEAs was performed in 3077 consecutive patients. The ratio of men to women was 2:1, and the ratio of patients who were asymptomatic to patients who were symptomatic was 2.3:1. The following in-hospital outcome data were analyzed: transient ischemic attack (TIA), stroke, mortality, combined TIA or stroke, and combined stroke or mortality. Univariate and multivariate analysis results of selected risk factors for an adverse perioperative event were assessed with generalized estimating equation analysis with backwards selection. The following risk factors were considered: gender, preoperative neurologic status, urgency of operation, type of arteriotomy repair, reoperative CEA for recurrent stenosis, history of cardiopulmonary disease, previous coronary artery intervention, simultaneous CEA and coronary bypass grafting surgery, renal failure, and diabetes. RESULTS: Univariate analysis results of gender differences revealed that women were at higher risk for a postoperative TIA or stroke (3.3% for women versus 2.1% for men; odds ratio [OR], 1.6; confidence interval [CI], 1.04 to 2.5; P =.03) and for postoperative stroke or mortality (3.1% for women versus 2.1% for men; OR, 1.6; CI, 1.04 to 2.5; P =.03). Multivariate analysis results showed that female gender was an independent predictor for a postoperative TIA or stroke (OR, 1.7; CI, 1.1 to 2.6; P =.03). Further analysis disclosed that women who were asymptomatic were at greater risk than were men for postoperative stroke or mortality (OR, 2.3; CI, 1.3 to 3.9; P =.003). Conversely, there was no gender association for postoperative stroke or mortality in the consideration of only patients who were symptomatic (OR, 1.0; CI, 0.45 to 2.1; P =.95). The interaction between women and preoperative symptoms approached significance (P =.07) with respect to postoperative stroke and mortality rate, which suggests that the gender effect could be influenced by the clinical presentation. CONCLUSION: The combined TIA or stroke and stroke or mortality rates are higher in women as compared with men in the postoperative period, but these risks remain acceptable when CEA is performed for appropriate indications. The interaction between symptoms and gender suggests that, in patients who are asymptomatic, women are more likely than are men to have early complications. However, there is no gender difference in patients who are symptomatic. Therefore, despite a low postoperative complication rate, CEA is appropriate in both women who are asymptomatic and women who are symptomatic only if the postoperative TIA, stroke, and mortality rates are appreciably lower than in the natural history of medical management of these patients. PMID- 11932675 TI - Cost of routine screening for carotid and lower extremity occlusive disease in patients with abdominal aortic aneurysms. AB - OBJECTIVE: The burden of clinically relevant noncoronary atherosclerotic occlusive disease in patients with abdominal aortic aneurysms (AAAs) is poorly defined. Furthermore, the cost-effectiveness of routine versus selective preoperative noninvasive examination of the carotid and lower extremity arterial beds has not been established in patients who undergo elective AAA repair. METHODS: Diagnostic vascular laboratory study results were reviewed in 206 patients who underwent evaluation before AAA repair from 1994 to 1998. The patients underwent routine preoperative carotid duplex scan examinations and lower extremity Doppler scan arterial studies with ankle-brachial index (ABI) determinations. The medical records were reviewed for the identification of clinical evidence consistent with cerebrovascular or lower extremity arterial occlusive disease. The costs of routine screening and selective screening were determined with Medicare reimbursement schedules. RESULTS: The prevalence rate of advanced (80% to 100%) carotid artery stenosis (CAS) was 3.4%, and 18% of the patients had CAS between 60% and 100%. Advanced peripheral vascular occlusive disease (PVOD; ABI, <0.3) was found in 3% of the patients, and 12% of the patients had an ABI of less than 0.6. Most patients with advanced CAS (71%) or advanced PVOD (83%) had clinical indications of their disease. The absence of clinical evidence of disease had a negative predictive value of 99% for both advanced CAS and PVOD. The cost of routine screening for all patients for advanced CAS was $5445 per case. Routine screening for severe PVOD costs were $3732 per case discovered. In contrast, the costs for selective screening for advanced CAS or PVOD in patients with appropriate history or symptoms were $1258 and $785 per case found, respectively. CONCLUSION: Routine noninvasive diagnostic testing for the identification of asymptomatic CAS and PVOD in patients with AAA may not be justified. Preoperative screening is more clearly indicated for patients with AAAs who have clinical evidence suggestive of CAS or PVOD. PMID- 11932676 TI - Intraoperative angioscopy may improve the outcome of in situ saphenous vein bypass grafting: a prospective study. AB - OBJECTIVE: To find out whether intraoperative angioscopic assistance has any effect on graft outcome in patients with critical leg ischemia. MATERIAL AND METHODS: One hundred one patients requiring a below-knee bypass were assigned to undergo in situ saphenous vein bypass with or without intraoperative angioscopic assistance; otherwise treated similarly including preoperative duplex vein mapping, intraoperative graft flow measurements, and angiography. Data on operative details, morbidity, hospital stay, and graft patency were collected prospectively and compared. All patients were followed up for 12 months. RESULTS: The group that underwent angioscopy (A) and the control group (B) were similar in all respects, except for the number of patients enrolled in the groups (32 and 69, respectively). Angioscopy revealed incompletely destructed valves in 34 patients (range, 0 to 5; mean 1), undiagnosed vein branches in 111 patients (mean 4.3), and partly occluding thrombus in 5 patients. The number of postoperative arteriovenous fistulas with signs of failing graft and a need for angiographic or surgical reintervention were significantly higher in group B (P <.0001). The 1 year primary patency rate was significantly better in group A (P <.01), but the primary assisted and secondary patency rates did not differ between the groups. CONCLUSIONS: Angioscopic assistance has an impact on primary graft patency, minimizes the risk for graft failure and thus reduces the need for reintervention by allowing identification of persistent saphenous vein branches, incomplete valve destruction, and partly occluding graft thrombus without adding extra operative time. PMID- 11932677 TI - Magnetic resonance angiography unmasks reliable target vessels for pedal bypass grafting in patients with diabetes mellitus. AB - OBJECTIVE: The purpose of this study was to evaluate the performance of pedal bypass grafts to foot vessels detected by magnetic resonance angiography (MRA) that were occult in conventional angiography in patients with diabetes mellitus and severe arterial occlusive disease. METHODS: Vascular surgery and radiology registries were reviewed for patients with pedal bypass grafts to arteries that were not detected with digital subtraction angiography but unmasked with MRA. From December 1997 to March 1999, 15 patients (mean age, 73 +/- 8 years) were identified and analyzed retrospectively. All the patients were diabetic, with 60% being insulin-dependent. Advanced tissue loss was the operative indication in all the cases. Distal anastomosis was performed to the dorsalis pedis artery in 10 cases and to the plantar artery in five cases, with ipsilateral greater saphenous vein as graft material in all the cases. RESULTS: The perioperative mortality rate was 7% (1 of 15 cases). One graft occlusion resulted in a secondary patency rate of 93.1%. During a mean follow-up examination period of 22 months, no graft occlusions and one major amputation were noted, which resulted in a secondary patency rate of 93.1% and a limb salvage rate of 89.5% at 36 months. CONCLUSION: Foot vessels that were occult in conventional angiography but could be detected with MRA were shown to be suitable target vessels for pedal bypass grafting with promising results. PMID- 11932678 TI - Long-term results of revised infrainguinal arterial reconstructions. AB - PURPOSE: Infrainguinal arterial reconstruction with vein as the conduit has been regarded as the gold standard within the past two decades. However, as many as 20% of patients undergoing these bypass grafting procedures may need secondary interventions for continued patency. Once these reconstructions have been altered, there is concern about the continued patency of these types of revascularizations. In this study, we evaluated the long-term patency of venous reconstructions that had been revised, compared them with those bypass grafting procedures that did not require any intervention, and analyzed long-term outcome. METHODS: In the past 15 years, 3944 infrainguinal arterial reconstructions were completed with vein as the conduit. A total of 2780 were performed with the vein in situ, and 1164 were performed by using excised vein in a single piece or spliced configuration. Indication, risk factors, and patient demographics were evaluated. Grafts were stratified into revised and unrevised, in situ and excised; excised vein was then stratified into spliced and non-spliced. All grafts were followed with duplex ultrasound scans performed at 3, 6, and 12 months in the first year and every 6 months thereafter. Statistical analysis was performed by using Gehan's generalized Wilcoxon test. RESULTS: Demographics and indications were similar between groups. Nine percent of all excised veins required some revision, compared with 10% of all in situ reconstructions. When the excised veins were further stratified, 6% of single-piece venous conduits were revised, as opposed to 14% of reconstructions with spliced venous reconstruction. The 5-year patency rates were 67% for revised reconstructions and 78% for unrevised reconstructions (P <.0001). The 5-year patency rate of unrevised in situ bypass grafts was 81%, as compared with 69% for revised in situ reconstruction (P <.0001), and the 5-year patency rate for unrevised excised veins was 68%, with revised excised vein having a 5-year patency rate of 59% (P = not significant). CONCLUSION: Venous conduits that require revision have a significantly lower long-term patency rate than those that were unrevised. Grafts that require revision may be best suited for aggressive surveillance protocol to maximize long-term patency. PMID- 11932679 TI - Limitations in the use of rifampicin-gelatin grafts against virulent organisms. AB - OBJECTIVE: Efficacy and duration of antibacterial activity of rifampicin-gelatin grafts against virulent organisms were evaluated in an animal model. MATERIALS AND METHODS: Rifampicin-gelatin grafts were prepared with impregnation of Gelseal (Vascutek Ltd, Scotland) graft in 1 mg/mL rifampicin solution. Rifampicin-gelatin grafts (6 cm long; n = 24) and plain Gelseal grafts as controls (n = 4) were implanted into the canine abdominal aorta with inoculation of Staphylococcus epidermidis, Escherichia coli, or methicillin-resistant Staphylococcus aureus (MRSA), and the rifampicin-gelatin grafts were retrieved after 1 to 4 weeks. Disks cut from the retrieved rifampicin-gelatin grafts were placed on agar plates streaked with one of the organisms, and the graft antibacterial activity was assessed with the width of the inhibition zone. RESULTS: In in vitro tests, initial inhibition zones (inhibition zone of 24 hours after incubation) of rifampicin-gelatin grafts against S epidermidis, MRSA, and E coli were 40.0 +/- 0.3 mm, 36.0 +/- 0.2 mm, and 11.8 +/- 0.1 mm, respectively. In the implantation, S epidermidis -inoculated rifampicin-gelatin grafts had no findings of graft infection, and no colony growth was recognized on the plates streaked with the perigraft fluids. Initial inhibition zones of S epidermidis -inoculated rifampicin-gelatin grafts retrieved at 1 or 2 weeks were 20.1 +/- 1.1 mm and 7.6 +/- 1.0 mm, respectively. In E coli -inoculated and MRSA-inoculated rifampicin gelatin grafts, all of the eight animals had perigraft abscess, and blood culture test results probed septicemia in five animals with patent grafts at death. Inhibition zones against E coli or MRSA were not formed on the plates streaked with the same organism, whereas initial inhibition zones of E coli -inoculated and MRSA-inoculated rifampicin-gelatin grafts on S epidermidis -streaked plates were 8.0 +/- 0.2 mm and 18.5 +/- 0.5 mm, respectively. In the MRSA group, however, recolonization of high minimal inhibitory concentration strains developed within the inhibition zones as early as 24 hours. Histologically, neither organisms nor inflammatory cells were found in S epidermidis -inoculated rifampicin-gelatin grafts and tissue ingrowth was recognized at 2 to 4 weeks, whereas E coli -inoculated and MRSA-inoculated rifampicin-gelatin grafts had aggressive neutrophil infiltration into the graft interstices, revealing establishment of uncontrollable graft infection. CONCLUSION: These results suggested that rifampicin-gelatin grafts are clearly valid for S epidermidis infection, whereas no efficacy was recognized against either MRSA or E coli graft infection because of early development of high minimal inhibitory concentration MRSA strains or poor susceptibility. PMID- 11932680 TI - Hepatocyte growth factor prevents intimal hyperplasia in rabbit carotid expanded polytetrafluoroethylene grafting. AB - PURPOSE: The major cause of vascular prosthesis failure is anastomotic intimal hyperplasia caused by the proliferation and migration of smooth muscle cells. Hepatocyte growth factor (HGF) is an endothelium-specific growth factor that exerts a mitogenic action on endothelial cells. This study was designed to examine the effect of HGF on the suppression of intimal hyperplasia after small caliber expanded polytetrafluoroethylene (ePTFE) grafting. METHODS: An ePTFE graft, 2 mm in diameter and 30 mm in length, was implanted in the left common carotid arteries of Japanese white rabbits, after which the animals were fed with a 1.0% cholesterol diet. HGF was infused intravenously immediately and then every day for 7 days at doses of 0.3 mg/body (the 0.3-mg HGF group; n = 20) or 1.0 mg/body (the 1.0-mg HGF group; n = 17). A control group (n = 20) underwent infusion with saline solution. The rabbits were killed on postoperative days (PODs) 1, 2, 3, 5, 7, and 28. RESULTS: The patency rates on POD 28 were 33%, 55%, and 100% in the control, the 0.3-mg HGF, and the 1.0-mg HGF groups, respectively, with a significant difference between the control and the 1.0-mg HGF group (P <.05). Endothelial-like cells were seen on the intraluminal surface of the graft only near the anastomotic site on POD 5 in the 1.0-mg HGF group. Intimal thickness at the distal anastomosis was 284 +/- 140 microm, 106 +/- 18 microm, and 67 +/- 10 microm in the control, the 0.3-mg HGF, and the 1.0-mg HGF groups, respectively, with a significant difference between the control and both HGF groups (P <.05). The number of anti-embryonic smooth muscle antibody positive cells at the distal anastomosis was 28.6 +/- 0.8, 3.8 +/- 2.8, and 3.9 +/- 0.9 in the control, the 0.3-mg HGF, and the 1.0-mg HGF groups, respectively, with a significant difference between the control and both HGF groups (P <.01). CONCLUSION: HGF might suppress intimal thickness at the anastomotic site and improve the patency rate via rapid reendothelialization by POD 28 in a rabbit carotid ePTFE grafting model. PMID- 11932681 TI - Reduced protease inhibitory capacity in patients with abdominal aortic aneurysms is reversed with surgical repair. AB - BACKGROUND: Abdominal aortic aneurysm (AAA) disease is a complex degenerative process that is associated with elevated proteolytic activity. This increased proteolytic activity may be linked to an imbalance in the protease regulatory mechanisms. We hypothesize that reduced AAA plasma inhibitory capacity is related to increased elastolytic activity observed in patients with AAA, which promotes matrix degradation in the aortic wall and AAA expansion. METHODS: Preoperative plasma from patients with AAA (n = 86; mean age, 75 +/- 8 years) and healthy controls (n = 30; mean age, 72 +/- 10 years) was assayed for inhibitory capacity. Postoperative plasma inhibitory capacity was evaluated with comparison of the activity of porcine pancreatic elastase on succinylated elastin substrate, in the presence and absence of AAA and control plasma. RESULTS: AAA plasma showed a significantly reduced capacity to inhibit porcine pancreatic elastase (71.31% +/- 1.47%) compared with control plasma (85.52% +/- 1.70%; P <.001). Within the AAA group, the inhibitory capacity increased with AAA diameter (P <.05). In addition, surgical repair of AAA resulted in a significant increase in plasma inhibitory capacity (79.83% +/- 1.86%) as compared with preoperative levels (63.98% +/- 2.69%; P <.001). Of note, the postoperative plasma inhibitory capacity is not significantly different than that of control. CONCLUSION: AAA plasma has a significantly reduced inhibitory capacity, which varies with aortic diameter and returns to control levels after surgical repair of the AAA. This suggests that protease regulation is a dynamic balance that changes with AAA progression and may play an important role in AAA progression. PMID- 11932682 TI - Rupture of abdominal aortic aneurysm with tear of inferior vena cava in a patient with prior endograft. AB - We report a case of contained rupture of abdominal aortic aneurysm and tear of the inferior vena cava (IVC) 15 months after placement of an aortic endograft (ANEURX graft, Medtronic, Sunnyvale, Calif). A 63-year-old man with significant coronary artery disease underwent endograft exclusion of abdominal aortic aneurysm with Aneurx graft. The patient was seen with a rupture of the aortic aneurysm, probably caused by poor proximal fixation of the graft associated with separation of the left iliac extension limb from the main body of the graft. Angulated right iliac limb of the stent graft penetrated into the Ivc just above the common iliac junction and caused sealed perforation. Successful repair with aortobiiliac graft reconstruction after removal of the endograft was accomplished. The IVC laceration was repaired. Possible mechanisms of failure of endograft are discussed. PMID- 11932683 TI - Nondissecting aneurysm of the thoracic aorta with arteritis in systemic lupus erythematosus. AB - A nondissecting aneurysm of the distal descending thoracic aorta, 55 mm in diameter, in a 35-year-old woman who was undergoing 24-year steroid therapy of systemic lupus erythematosus was successfully resected and replaced with a tube graft. The suture line of the aortic wall was reinforced with a polytetrafluoroethylene felt strip. Histologically, the media and adventitia contained numerous lymphoplasmacytic infiltrates that consisted of B and activated T cells mixed with plasmacytes. These findings would have been in accordance with active aortitis. Systemic lupus erythematosus was suggested as possibly associated with nondissecting thoracic aortic aneurysm, especially in patients undergoing chronic steroid therapy in which inflammation continued. PMID- 11932684 TI - Superior mesenteric artery pseudoaneurysm successfully treated with polytetrafluoroethylene covered stent. AB - A postoperative superior mesenteric artery pseudoaneurysm that communicates with a pancreatic pseudocyst after aortic surgery is a difficult management problem. Untreated, this condition can lead to exsanguination. Traditional surgical treatment has many potential complications. Endovascular repair has the potential for avoidance of surgical complications. We present the first superior mesenteric artery pseudoaneurysm successfully treated with A polytetrafluorethylene covered stent. PMID- 11932685 TI - Successful bilateral transaortic renal endarterectomy after failed renal artery angioplasty and stenting: a case report. AB - Restenosis after renal angioplasty and stenting is usually treated with repeat angioplasty or surgery. Because development of substantial postangioplasty periarterial fibrosis is thought to preclude transaortic endarterectomy, renal artery bypass grafting has been the operation of choice in this setting. This report describes successful bilateral transaortic renal thromboendarterectomy undertaken for the treatment of restenosis after percutaneous angioplasty and stenting for renovascular hypertension caused by bilateral renal artery nonostial stenosis. PMID- 11932686 TI - Treatment of abdominal aortic anastomotic pseudoaneurysm with percutaneous coil embolization. AB - Intraabdominal anastomotic pseudoaneurysms continue to be a late complication of aortic reconstructive procedures. Early surgical repair is critical but is associated with high operative mortality rates. We present a patient who was diagnosed with a distal anastomotic pseudoaneurysm 13 months after transabdominal repair of a symptomatic abdominal aortic aneurysm. Because of the poor operative risk, the patient was considered for a less invasive approach and underwent coil embolization of the abdominal aortic anastomotic pseudoaneurysm. The patient remains without recurrence of pseudoaneurysm 3.5 years later. PMID- 11932687 TI - Retroperitoneoscopic lumbar sympathectomy. AB - We present a series of 27 consecutive unselected patients who underwent 29 retroperitoneoscopic lumbar sympathectomies. There were 21 male patients and six female patients, with a mean age of 45 years (RANGE, 21 to 28 years). Twenty-two patients had ischemia of the lower limb, and five patients had severe reflex sympathetic dystrophy. The retroperitoneal space was developed with a balloon trocar inserted through a small incision in the flank. Additional trocars used for endoscopic instruments. The sympathetic chain from the ganglia second lumbar vertebrae to the fourth lumbar vertebrae was resected. The procedure was successfully accomplished in all the patients without any operative or postoperative complications. The mean operative time Was 136 minutes, and the mean hospital stay was 1.4 days. All the patients had significant improvement of pain or dystrophic changes. Retroperitoneoscopic lumbar sympathectomy successfully combines the advantages of minimally invasive surgery with the effectiveness of the open procedure. PMID- 11932688 TI - A proposal for revitalization: an international society for vascular intervention. PMID- 11932689 TI - Regarding "Pathogenetic heterogeneity of in-stent lesion formation in human peripheral arterial disease". PMID- 11932690 TI - Regarding "Limitations in the use of rifampicin-gelatin grafts against virulent organisms". PMID- 11932691 TI - Surgeon-industry relationships: ethically responsible management of conflicts of interest. PMID- 11932692 TI - Regarding "Isolation of endothelial cells and their progenitor cells from human peripheral blood". PMID- 11932694 TI - Regarding "Bedside vena cava filter placement guided with intravascular ultrasound". PMID- 11932696 TI - Regarding "High endogenous estradiol is associated with increased venous distensibility and clinical evidence of varicose veins in menopausal women". PMID- 11932698 TI - Regarding "Early results of infragenicular revascularization based solely on duplex arteriography". PMID- 11932703 TI - Prevention remains the best medicine. PMID- 11932702 TI - Tearing down the barriers. PMID- 11932704 TI - Media studies for scientists. PMID- 11932705 TI - Papers square up over potential Pulitzer for cancer-centre critics. PMID- 11932707 TI - Online tumour bank aims to offer ready route to tissues. PMID- 11932709 TI - Charity launches not-for-profit drug industry. PMID- 11932711 TI - US physicists unite behind big ideas. PMID- 11932710 TI - Sparks fly as electrostatic facility fizzles out. PMID- 11932712 TI - University research to touch base with NASA. PMID- 11932714 TI - On the offensive. PMID- 11932716 TI - Communication should not be left to scientists. PMID- 11932715 TI - Miller revealed new ways to study the origins of life. PMID- 11932722 TI - Toxicology: a risky business. PMID- 11932724 TI - Astrophysics: the evidence in the afterglow. PMID- 11932723 TI - Medicine: danger--misfolding proteins. PMID- 11932725 TI - Stem cells: cell fusion causes confusion. PMID- 11932727 TI - Biogeography: baffled over bison. PMID- 11932728 TI - Telecommunications: a boost for fibre optics. PMID- 11932729 TI - Genetics: immaculate misconception. PMID- 11932730 TI - Developmental biology: the plastic face. PMID- 11932732 TI - Microfabrication technology: organized assembly of carbon nanotubes. AB - Nanoscale structures need to be arranged into well-defined configurations in order to build integrated systems. Here we use a chemical-vapour deposition method with gas-phase catalyst delivery to direct the assembly of carbon nanotubes in a variety of predetermined orientations onto silicon/silica substrates, building them into one-, two- and three-dimensional arrangements. The preference of nanotubes to grow selectively on and normal to silica surfaces forces them to inherit the lithographically machined template topography of their substrates, allowing the sites of nucleation and the direction of growth to be controlled. PMID- 11932733 TI - Sperm competition: motility and the midpiece in primates. AB - In animals with multiple-partner mating systems, the gametes of two or more males must compete to fertilize a given set of ova. Here we show that the volume of the midpiece in individual sperm is significantly greater in primate species in which the females mate with multiple partners, and in which males have larger testes in relation to their body weight, than in those species that mate with only one partner and have relatively small testes. Our results indicate that sexual selection by sperm competition has influenced the evolution of a specific component of male-gamete morphology, the volume of the sperm midpiece. PMID- 11932734 TI - Aerosol formation: atmospheric particles from organic vapours. AB - Aerosol particles produced over forested areas may affect climate by acting as nuclei for cloud condensation, but their composition (and hence the chemical species that drive their production) remains an open question. Here we show, to our knowledge for the first time, that these newly formed particles (3-5 nm in diameter) are composed primarily of organic species, such as cis-pinonic acid and pinic acid, produced by oxidation of terpenes in organic vapours released from the canopy. PMID- 11932735 TI - Nanomechanics: response of a strained semiconductor structure. AB - The nanomechanical properties of thin silicon films will become increasingly critical in semiconductor devices, particularly in the context of substrates that consist of a silicon film on an insulating layer (known as silicon-on-insulator, or SOI, substrates). Here we use very small germanium crystals as a new type of nanomechanical stressor to demonstrate a surprising mechanical behaviour of the thin layer of silicon in SOI substrates, and to show that there is a large local reduction in the viscosity of the oxide on which the silicon layer rests. These findings have implications for the use of SOI substrates in nanoelectronic devices. PMID- 11932736 TI - An extensive network of coupling among gene expression machines. AB - Gene expression in eukaryotes requires several multi-component cellular machines. Each machine carries out a separate step in the gene expression pathway, which includes transcription, several pre-messenger RNA processing steps and the export of mature mRNA to the cytoplasm. Recent studies lead to the view that, in contrast to a simple linear assembly line, a complex and extensively coupled network has evolved to coordinate the activities of the gene expression machines. The extensive coupling is consistent with a model in which the machines are tethered to each other to form 'gene expression factories' that maximize the efficiency and specificity of each step in gene expression. PMID- 11932738 TI - The signature of supernova ejecta in the X-ray afterglow of the gamma-ray burst 011211. AB - Now that gamma-ray bursts (GRBs) have been determined to lie at cosmological distances, their isotropic burst energies are estimated to be as high as 1054 erg (ref. 2), making them the most energetic phenomena in the Universe. The nature of the progenitors responsible for the bursts remains, however, elusive. The favoured models range from the merger of two neutron stars in a binary system to the collapse of a massive star. Spectroscopic studies of the afterglow emission could reveal details of the environment of the burst, by indicating the elements present, the speed of the outflow and an estimate of the temperature. Here we report an X-ray spectrum of the afterglow of GRB011211, which shows emission lines of magnesium, silicon, sulphur, argon, calcium and possibly nickel, arising in metal-enriched material with an outflow velocity of the order of one-tenth the speed of light. These observations strongly favour models where a supernova explosion from a massive stellar progenitor precedes the burst event and is responsible for the outflowing matter. PMID- 11932737 TI - Inherent toxicity of aggregates implies a common mechanism for protein misfolding diseases. AB - A range of human degenerative conditions, including Alzheimer's disease, light chain amyloidosis and the spongiform encephalopathies, is associated with the deposition in tissue of proteinaceous aggregates known as amyloid fibrils or plaques. It has been shown previously that fibrillar aggregates that are closely similar to those associated with clinical amyloidoses can be formed in vitro from proteins not connected with these diseases, including the SH3 domain from bovine phosphatidyl-inositol-3'-kinase and the amino-terminal domain of the Escherichia coli HypF protein. Here we show that species formed early in the aggregation of these non-disease-associated proteins can be inherently highly cytotoxic. This finding provides added evidence that avoidance of protein aggregation is crucial for the preservation of biological function and suggests common features in the origins of this family of protein deposition diseases. PMID- 11932739 TI - Bunching of fractionally charged quasiparticles tunnelling through high-potential barriers. AB - Shot noise measurements have been used to measure the charge of quasiparticles in the fractional quantum Hall (FQH) regime. To induce shot noise in an otherwise noiseless current of quasiparticles, a barrier is placed in its path to cause weak backscattering. The measured shot noise is proportional to the charge of the quasiparticles; for example, at filling factor v=1/3, noise corresponding to q=e/3 appears. For increasingly opaque barriers, the measured charge increases monotonically, approaching q=e asymptotically. It was therefore believed that only electrons, or alternatively, three bunched quasiparticles, can tunnel through high-potential barriers encountered by a noiseless current of quasiparticles. Here we investigate the interaction of e/3 quasiparticles with a strong barrier in FQH samples and find that bunching of quasiparticles in the strong backscattering limit depends on the average dilution of the quasiparticle current. For a very dilute current, bunching ceases altogether and the transferred charge approaches q=e/3. This surprising result demonstrates that quasiparticles can tunnel individually through high-potential barriers originally thought to be opaque for them. PMID- 11932740 TI - Atomic-scale images of charge ordering in a mixed-valence manganite. AB - Transition-metal perovskite oxides exhibit a wide range of extraordinary but imperfectly understood phenomena. The best known examples are high-temperature superconductivity in copper oxides, and colossal magnetoresistance in manganese oxides ('manganites'). All of these materials undergo a range of order-disorder transitions associated with changes in charge, spin, orbital and lattice degrees of freedom. Measurements of such order are usually made by diffraction techniques, which detect the ionic cores and the spins of the conduction electrons. Unfortunately, because such techniques are only weakly sensitive to valence electrons and yield superpositions of signals from distinct submicrometre scale phases, they cannot directly image phase coexistence and charge ordering, two key features of the manganites. Here we present scanning tunnelling microscope measurements of the manganite Bi1-xCaxMnO3. We show that charge ordering and phase separation can be resolved in real space with atomic-scale resolution. By taking together images and current-voltage spectroscopy data we find that charge order correlates with both structural order and the local conductive state (either metallic or insulating). These experiments provide an atomic-scale basis for descriptions of manganites as mixtures of electronically and structurally distinct phases. PMID- 11932741 TI - Abiogenic formation of alkanes in the Earth's crust as a minor source for global hydrocarbon reservoirs. AB - Natural hydrocarbons are largely formed by the thermal decomposition of organic matter (thermogenesis) or by microbial processes (bacteriogenesis). But the discovery of methane at an East Pacific Rise hydrothermal vent and in other crustal fluids supports the occurrence of an abiogenic source of hydrocarbons. These abiogenic hydrocarbons are generally formed by the reduction of carbon dioxide, a process which is thought to occur during magma cooling and-more commonly-in hydrothermal systems during water-rock interactions, for example involving Fischer-Tropsch reactions and the serpentinization of ultramafic rocks. Suggestions that abiogenic hydrocarbons make a significant contribution to economic hydrocarbon reservoirs have been difficult to resolve, in part owing to uncertainty in the carbon isotopic signatures for abiogenic versus thermogenic hydrocarbons. Here, using carbon and hydrogen isotope analyses of abiogenic methane and higher hydrocarbons in crystalline rocks of the Canadian shield, we show a clear distinction between abiogenic and thermogenic hydrocarbons. The progressive isotopic trends for the series of C1-C4 alkanes indicate that hydrocarbon formation occurs by way of polymerization of methane precursors. Given that these trends are not observed in the isotopic signatures of economic gas reservoirs, we can now rule out the presence of a globally significant abiogenic source of hydrocarbons. PMID- 11932742 TI - Long-lived vortices as a mode of deep ventilation in the Greenland Sea. AB - The Greenland Sea is one of a few sites in the world ocean where convection to great depths occurs-a process that forms some of the densest waters in the ocean. But the role of deep convective eddies, which result from surface cooling and mixing across density surfaces followed by geostrophic adjustment, has not been fully taken into account in the description of the initiation and growth of convection. Here we present tracer, float and hydrographic observations of long lived ( approximately 1 year) and compact ( approximately 5 km core diameter) vortices that reach down to depths of 2 km. The eddies form in winter, near the rim of the Greenland Sea central gyre, and rotate clockwise with periods of a few days. The cores of the observed eddies are constituted from a mixture of modified Atlantic water that is warm and salty with polar water that is cold and fresh. We infer that these submesoscale coherent eddies contribute substantially to the input of Atlantic and polar waters to depths greater than 500 m in the central Greenland Sea. PMID- 11932743 TI - Fin development in a cartilaginous fish and the origin of vertebrate limbs. AB - Recent fossil finds and experimental analysis of chick and mouse embryos highlighted the lateral fin fold theory, which suggests that two pairs of limbs in tetrapods evolved by subdivision of an elongated single fin. Here we examine fin development in embryos of the primitive cartilaginous fish, Scyliorhinus canicula (dogfish) using scanning electron microscopy and investigate expression of genes known to be involved in limb positioning, identity and patterning in higher vertebrates. Although we did not detect lateral fin folds in dogfish embryos, Engrailed-1 expression suggests that the body is compartmentalized dorso ventrally. Furthermore, specification of limb identity occurs through the Tbx4 and Tbx5 genes, as in higher vertebrates. In contrast, unlike higher vertebrates, we did not detect Shh transcripts in dogfish fin-buds, although dHand (a gene involved in establishing Shh) is expressed. In S. canicula, the main fin axis seems to lie parallel to the body axis. 'Freeing' fins from the body axis and establishing a separate 'limb' axis has been proposed to be a crucial step in evolution of tetrapod limbs. We suggest that Shh plays a critical role in this process. PMID- 11932744 TI - The cost of inbreeding in Arabidopsis. AB - Population geneticists have long sought to estimate the distribution of selection intensities among genes of diverse function across the genome. Only recently have DNA sequencing and analytical techniques converged to make this possible. Important advances have come from comparing genetic variation within species (polymorphism) with fixed differences between species (divergence). These approaches have been used to examine individual genes for evidence of selection. Here we use the fact that the time since species divergence allows combination of data across genes. In a comparison of amino-acid replacements among species of the mustard weed Arabidopsis with those among species of the fruitfly Drosophila, we find evidence for predominantly beneficial gene substitutions in Drosophila but predominantly detrimental substitutions in Arabidopsis. We attribute this difference to the Arabidopsis mating system of partial self-fertilization, which corroborates a prediction of population genetics theory that species with a high frequency of inbreeding are less efficient in eliminating deleterious mutations owing to their reduced effective population size. PMID- 11932745 TI - Naturally secreted oligomers of amyloid beta protein potently inhibit hippocampal long-term potentiation in vivo. AB - Although extensive data support a central pathogenic role for amyloid beta protein (Abeta) in Alzheimer's disease, the amyloid hypothesis remains controversial, in part because a specific neurotoxic species of Abeta and the nature of its effects on synaptic function have not been defined in vivo. Here we report that natural oligomers of human Abeta are formed soon after generation of the peptide within specific intracellular vesicles and are subsequently secreted from the cell. Cerebral microinjection of cell medium containing these oligomers and abundant Abeta monomers but no amyloid fibrils markedly inhibited hippocampal long-term potentiation (LTP) in rats in vivo. Immunodepletion from the medium of all Abeta species completely abrogated this effect. Pretreatment of the medium with insulin-degrading enzyme, which degrades Abeta monomers but not oligomers, did not prevent the inhibition of LTP. Therefore, Abeta oligomers, in the absence of monomers and amyloid fibrils, disrupted synaptic plasticity in vivo at concentrations found in human brain and cerebrospinal fluid. Finally, treatment of cells with gamma-secretase inhibitors prevented oligomer formation at doses that allowed appreciable monomer production, and such medium no longer disrupted LTP, indicating that synaptotoxic Abeta oligomers can be targeted therapeutically. PMID- 11932746 TI - A global disorder of imprinting in the human female germ line. AB - Imprinted genes are expressed differently depending on whether they are carried by a chromosome of maternal or paternal origin. Correct imprinting is established by germline-specific modifications; failure of this process underlies several inherited human syndromes. All these imprinting control defects are cis-acting, disrupting establishment or maintenance of allele-specific epigenetic modifications across one contiguous segment of the genome. In contrast, we report here an inherited global imprinting defect. This recessive maternal-effect mutation disrupts the specification of imprints at multiple, non-contiguous loci, with the result that genes normally carrying a maternal methylation imprint assume a paternal epigenetic pattern on the maternal allele. The resulting conception is phenotypically indistinguishable from an androgenetic complete hydatidiform mole, in which abnormal extra-embryonic tissue proliferates while development of the embryo is absent or nearly so. This disorder offers a genetic route to the identification of trans-acting oocyte factors that mediate maternal imprint establishment. PMID- 11932747 TI - Bone marrow cells adopt the phenotype of other cells by spontaneous cell fusion. AB - Recent studies have demonstrated that transplanted bone marrow cells can turn into unexpected lineages including myocytes, hepatocytes, neurons and many others. A potential problem, however, is that reports discussing such 'transdifferentiation' in vivo tend to conclude donor origin of transdifferentiated cells on the basis of the existence of donor-specific genes such as Y-chromosome markers. Here we demonstrate that mouse bone marrow cells can fuse spontaneously with embryonic stem cells in culture in vitro that contains interleukin-3. Moreover, spontaneously fused bone marrow cells can subsequently adopt the phenotype of the recipient cells, which, without detailed genetic analysis, might be interpreted as 'dedifferentiation' or transdifferentiation. PMID- 11932748 TI - Changing potency by spontaneous fusion. AB - Recent reports have suggested that mammalian stem cells residing in one tissue may have the capacity to produce differentiated cell types for other tissues and organs 1-9. Here we define a mechanism by which progenitor cells of the central nervous system can give rise to non-neural derivatives. Cells taken from mouse brain were co-cultured with pluripotent embryonic stem cells. Following selection for a transgenic marker carried only by the brain cells, undifferentiated stem cells are recovered in which the brain cell genome has undergone epigenetic reprogramming. However, these cells also carry a transgenic marker and chromosomes derived from the embryonic stem cells. Therefore the altered phenotype does not arise by direct conversion of brain to embryonic stem cell but rather through spontaneous generation of hybrid cells. The tetraploid hybrids exhibit full pluripotent character, including multilineage contribution to chimaeras. We propose that transdetermination consequent to cell fusion 10 could underlie many observations otherwise attributed to an intrinsic plasticity of tissue stem cells 9. PMID- 11932749 TI - DNMT1 and DNMT3b cooperate to silence genes in human cancer cells. AB - Inactivation of tumour suppressor genes is central to the development of all common forms of human cancer. This inactivation often results from epigenetic silencing associated with hypermethylation rather than intragenic mutations. In human cells, the mechanisms underlying locus-specific or global methylation patterns remain unclear. The prototypic DNA methyltransferase, Dnmt1, accounts for most methylation in mouse cells, but human cancer cells lacking DNMT1 retain significant genomic methylation and associated gene silencing. We disrupted the human DNMT3b gene in a colorectal cancer cell line. This deletion reduced global DNA methylation by less than 3%. Surprisingly, however, genetic disruption of both DNMT1 and DNMT3b nearly eliminated methyltransferase activity, and reduced genomic DNA methylation by greater than 95%. These marked changes resulted in demethylation of repeated sequences, loss of insulin-like growth factor II (IGF2) imprinting, abrogation of silencing of the tumour suppressor gene p16INK4a, and growth suppression. Here we demonstrate that two enzymes cooperatively maintain DNA methylation and gene silencing in human cancer cells, and provide compelling evidence that such methylation is essential for optimal neoplastic proliferation. PMID- 11932752 TI - The lactate dehydrogenases in malignant and non-malignant diseases. AB - The objective of this paper was to determine different patterns of Lactate Dehydrogenase(LD) isoenzymes in serum, pleural effusions and ascitic fluid collections and their usefulness in the differential diagnosis of malignant and non-malignant diseases. It was a case control study carried out at the Lagos University Teaching Hospital. Forty- six patients with pleural or ascitic fluid collection (male and female, age range between 18 and 65 years) made up of Twenty two patients with malignant diseases and twenty-four with non- malignant diseases were recruited. Control group comprised 50 apparently healthy subjects of both sexes with age ranging between 18 and 65 years. Serum, pleural and ascitic fluid collections were assayed for total lactate dehydrogenase(LD) and it s isoenzymes. Total LD was estimated using the pyruvate to lactate reaction. LD isoenzyme analysis was estimated by cellulose acetate electrophoresis and stained with formazan. The mean total serum LD activity in the disease groups (malignant and non-malignant) was significantly higher than the mean total serum LD activity in the control group. The pleural fluid LD was highest in patients with empyema thoracis. Five isoenzymes of LD (LD1 - LD5) were present in both control and patient sera. The serum LD1 and LD2 isoenzymes were predominant in the controls. The serum LD4 and LD5 isoenzymes were predominant in the disease groups (malignant and non-malignant). LD4 and LD5 were the predominant isoenzymes in both pleural and ascitic fluids obtained in malignant and non-malignant diseases. Serum LD4 was significantly higher than serum LD5 in non-malignant disease while serum LD5 was significantly higher than serum LD4 in malignant disease. The types of malignancies could not be differentiated by serum and effusion fluid LD isoenzyme pattern. Pleural fluid total LD estimation is useful in monitoring patients on chest tubes and may be used to determine when to insert them. Serum LD4 and LD5 differentiates malignant from non-malignant disease but the effusion LD isoenzyme pattern does not. PMID- 11932750 TI - p63 and p73 are required for p53-dependent apoptosis in response to DNA damage. AB - The tumour-suppressor gene p53 is frequently mutated in human cancers and is important in the cellular response to DNA damage. Although the p53 family members p63 and p73 are structurally related to p53, they have not been directly linked to tumour suppression, although they have been implicated in apoptosis. Given the similarity between this family of genes and the ability of p63 and p73 to transactivate p53 target genes, we explore here their role in DNA damage-induced apoptosis. Mouse embryo fibroblasts deficient for one or a combination of p53 family members were sensitized to undergo apoptosis through the expression of the adenovirus E1A oncogene. While using the E1A system facilitated our ability to perform biochemical analyses, we also examined the functions of p63 and p73 using an in vivo system in which apoptosis has been shown to be dependent on p53. Using both systems, we show here that the combined loss of p63 and p73 results in the failure of cells containing functional p53 to undergo apoptosis in response to DNA damage. PMID- 11932754 TI - Anaesthesia as a career--the influence of undergraduate education in a Nigerian Medical School. AB - Sixty-seven final year medical students of the Obafemi Awolowo University, Ile Ife, Nigeria were questioned to find out the influence of undergraduate education on their choice of future career. Eighty per cent of them found anaesthesia interesting and important but none would pick anaesthesia as a first choice for future career. One-third of the students studied did not even consider it. The students made several suggestions on how to improve undergraduate medical training. The author also suggests measures for general improvement in all aspects of anaesthetic practice in Nigeria with a view to attracting more residents into this specialty. PMID- 11932753 TI - Carriage rate of hepatitis-B surface antigen (HBsAg) in an urban community in Jos, Plateau State, Nigeria. AB - The carriage rate of Hepatitis-B surface antigen (HBsAg) in an urban community in Jos, the Plateau State capital, was studied to obtain the pattern of Hepatitis beta virus (HBV) spread within the community. HBsAg screening was performed on a consecutive sample of 524 apparently healthy individuals (293 males and 231 females) aged 15-65 years who voluntarily turned up for the survey. Fifty-four (10.3%) were HBsAg positive by ELISA. The carriage rate in females 30/231 (13.0%) was significantly higher than in the males 24/293 (8.2%) (p<0.05). In relation to age, 14/144 (9.7%) were aged <20 years, 11/121 (9.1%) were 21-30 years, 7/115 (6.1%) were 31-40 years, 14/89 (15.7%) were 41-50 years, 2/27 (7.4%) were 51-60 years and 6/28 (21.4%) were >61 years. In relation to marital status, carriage rate was highest among the divorced/widowed group (12.5%) followed by the married group (10.6%). Carriage rates in relation to occupation showed that infection was highest among traders (13.7%) and students (13.2%). The survey therefore confirms the endemicity of HBV infection in Jos and describes the groups that are at risk. This calls for health education of the general population on preventive measures to check the spread of the virus in the community. PMID- 11932755 TI - Episiotomies in Nigeria--should their use be restricted? AB - Episiotomy continues to be a frequently used procedure in obstetrics despite little scientific support for its routine use. The incidence of episiotomy and perineal tears and also the indications for episiotomy were therefore investigated in 1007 singleton deliveries between January 1, 1997 and December 31, 1997 at the Lagos University Teaching Hospital. The incidence of episiotomy was 54.9% of all deliveries. Episiotomy was more frequently performed in primipara (90.4%). Perineal tears occurred in only 18.8% of all deliveries, with majority in the multipara (93.6%). Most of the team were of first degree (98.4%). No third degree tear was recorded. The commonest indication for episiotomy was that of protecting the perineum from possible tears (80.7%). All the episiotomies were mediolateral. Episiotomy rate was higher among deliveries conducted by doctors. Performance of episiotomy had a positive correlation with increasing foetal weight. No major complication or maternal death attributable to episiotomy was recorded. The results of our study suggest that the use of episiotomy can be restricted to specified indications like instrumental and big babies delivery. It is also useful for the prevention of maternal morbidity through perineal laceration. Randomized control trials will however be necessary to clarify the controversies relating to restrictive episiotomy. PMID- 11932756 TI - A pathological review of intracranial tumours seen at the University College Hospital, Ibadan between 1980 and 1990. AB - The pattern of intracranial neoplasms in Ibadan Southwestern Nigeria appears to have evolved over three decades since the establishment of neurosurgical facility in 1962. Metastatic tumours to the intracranial compartment were the commonest neoplasms (23%); choriocarcinoma and multicentric involvement of the brain in Burkitt s lymphoma accounted for the great majority of the metastases. Well differentiated glial tumours are now being increasingly recognized in Ibadan, Nigeria. Glial tumours accounted for 20% of neoplasms, second in frequency to metastases. Pituitary adenomas and meningiomas accounted for 17.1% and 11.4% respectively. Germ cell tumours are however uncommon in Ibadan Nigeria having a low frequency (1.4%). A notable difference exists in the tumours found in children and adults. Astrocytomas accounted for the largest single group of neoplasms in children while metastatic tumours are the dominant group in adults. Continued improvement in neurosurgical facilities may improve diagnosis and cause more tumours to be verified histologically. This will contribute to the evolving pattern seen in Ibadan Southwestern Nigeria. PMID- 11932757 TI - Landau-Kleffner syndrome (acquired epileptic aphasia) in a Nigerian child. AB - Epilepsy is one of the commonest disorders presenting the neurological clinics in developing countries. There are attendant complications especially in untreated or poorly treated cases. In this article, we report an uncommon complication known as Landau-Kleffner syndrome in a Nigerian girl. The syndrome, typically seen in children consists of an acquired aphasia frequently associated with clinical seizures and paroxysmal electroencephalographic (EEG) accompaniment. There had been no adequate neuropathlogical correlations to explain the symptomatology; thus, various forms of pathogenesis have been ascribed to the syndrome. The clinical seizure is easily controlled by an antiepileptic; but speech recovery is variable, and this is usually poor in cases presenting one or two years after onset. PMID- 11932758 TI - Intralingual foregut duplication cyst: a case report. AB - A 20-month old girl presented with an intralingual cyst from birth, which interfered with feeding but not respiration. The cyst was completely excised and histology of the wall showed an epithelial lining of stratified squamous and respiratory type epithelium. There has been no recurrence and function of the tongue has remained normal. PMID- 11932760 TI - Fatal head injuries: the Lagos University Teaching Hospital experience (1993 1997). AB - Head injuries are a universal problem and they constitute a major public health problem both in terms of morbidity and mortality. The aim of this study is to examine the causes and pattern of fatal head injuries in the Lagos University Teaching Hospital (LUTH). A retrospective autopsy study of fatal head injuries in LUTH between January 1993 and December 1997 was done by retrieving autopsy reports, extracting relevant information and analysing them. Two hundred and fourteen such cases were reviewed (7% of all autopsies during that period). Majority were consequences of road traffic accidents (52%) and assault (12%). Adults between the ages of 21 and 40 years were the most affected (44%), followed by children 10 years and below (17%). Seventy seven percent of all cases were males. Common lesions identified include skull fractures (73%), intracranial haemorrhages (69%), lacerations and contusions (31%). PMID- 11932759 TI - Side-effects of drugs used in directly observed treatment short-course in newly diagnosed pulmonary tuberculosis subjects in Nigerians: a controlled clinical study. AB - A clinical study was carried out between January 1996 and December 1997 in Iwo Nigeria to determine among others the types and frequency of occurrence of side effects of anti-tuberculous drugs in the 8-month regimen recommended by the Nigerian National Tuberculosis Control Programme Committee (NTCP) for newly diagnosed (ND) Pulmonary Tuberculosis (PTB) cases. This was compared with a 6 month regimen used within the same population of ND PTB subjects at Iwo Nigeria. 65 (13%) of the 500 patients in group one on the 8-month regimen had various side effects while 7 (14%) of the 50 patients in group two on the 6-month regimen manifested various side-effects. The side-effects rate and frequencies were similar and in both, they were mild and did not materially lead to major change of the anti-tuberculous medications. The regimen used in this study in group one (NTCP recommended) and group two recorded low incidence of side-effects. It confirmed that Directly Observed Treatment Short-course (DOTS) when applied with the recommended regimen by the Nigerian NTCP is effective and is characterised by low and minor side-effects. PMID- 11932761 TI - Evaluation of a locally-made urease test for detecting Helicobacter pylori infection. AB - We studied the efficacy of a home-made urease test (HUT) in the detection of Helicobacter pylori (HP) infection in patients undergoing upper gastrointestinal endoscopy. In the first phase of the study, two antral biopsies each were obtained from 43 patients for urease tests using the CLOtest and a home-made buffered 2% urea solution with phenol red as indicator at pH 6.8 (2% HUT). Twenty six patients (60.5%) were HP positive, both by the 2% HUT and CLOtest with 100% concordance. In the second phase of the study three antral biopsies each and blood were obtained from 42 patients for the determination of HP status using a 10% HUT and a combination of culture and serology. Twenty-three patients (54.8%) were HP positive using the 10% HUT, while 32 patients (76.2%) were positive using the combination of 2 tests. Compared to this the sensitivity and specificity of the 10% HUT were 72% and 100% respectively. The CLOtest produced a colour change in a shorter time than the 2% and 10% HUT (median 1 hour versus 10 hours versus 16 hours p < 0.0001). In the third phase of the study, we observed that by doubling the biopsy size, the time required to obtain a colour change was significantly reduced (median 4.5 hours versus 10 hours p < 0.05). The HUT is easy to prepare, cheap, sufficiently sensitive and it is reliable enough to start treatment when positive. With 100% concordance and 1% the cost per test when compared to the commercially available CLOtest; the 10% HUT is hereby recommended for the detection of UP infection in our region. PMID- 11932766 TI - The gifts that keep on giving: physiological functions and evolutionary dynamics of male seminal proteins in Drosophila. AB - During mating, males transfer seminal proteins and peptides, along with sperm, to their mates. In Drosophila melanogaster, seminal proteins made in the male's accessory gland stimulate females' egg production and ovulation, reduce their receptivity to mating, mediate sperm storage, cause part of the survival cost of mating to females, and may protect reproductive tracts or gametes from microbial attack. The physiological functions of these proteins indicate that males provide their mates with molecules that initiate important reproductive responses in females. A new comprehensive EST screen, in conjunction with earlier screens, has identified approximately 90% of the predicted secreted accessory gland proteins (Acps). Most Acps are novel proteins and many appear to be secreted peptides or prohormones. Acps also include modification enzymes such as proteases and their inhibitors, and lipases. An apparent prohormonal Acp, ovulin (Acp26Aa) stimulates ovulation in mated Drosophila females. Another male-derived protein, the large glycoprotein Acp36DE, is needed for sperm storage in the mated female and through this action can also affect sperm precedence, indirectly. A third seminal protein, the protease inhibitor Acp62F, is a candidate for contributing to the survival cost of mating, given its toxicity in ectopic expression assays. That male-derived molecules manipulate females in these ways can result in a molecular conflict between the sexes that can drive the rapid evolution of Acps. Supporting this hypothesis, an unusually high fraction of Acps show signs consistent with their being targets of positive Darwinian selection. PMID- 11932767 TI - Plant sex determination and sex chromosomes. AB - Sex determination systems in plants have evolved many times from hermaphroditic ancestors (including monoecious plants with separate male and female flowers on the same individual), and sex chromosome systems have arisen several times in flowering plant evolution. Consistent with theoretical models for the evolutionary transition from hermaphroditism to monoecy, multiple sex determining genes are involved, including male-sterility and female-sterility factors. The requirement that recombination should be rare between these different loci is probably the chief reason for the genetic degeneration of Y chromosomes. Theories for Y chromosome degeneration are reviewed in the light of recent results from genes on plant sex chromosomes. PMID- 11932768 TI - Maternal-offspring conflict leads to the evolution of dominant zygotic sex determination. AB - Sex determination in many species involves interactions among maternally expressed genes (eg, mRNA's and proteins placed into the egg) and zygotically expressed genes. Recent studies have proposed that conflicting selective pressures can occur between maternally and zygotically expressed sex determining loci and that these may play a role in shaping the evolution of sex determining systems. Here we show that such genetic conflict occurs under very general circumstances. Whenever sex ratio among progeny in a family affects the fitness of either progeny in that family or maternal fitness, then maternal-zygotic genetic conflict occurs. Furthermore, we show that this conflict typically results in a "positive feedback loop" that leads to the evolution of a dominant zygotic sex determining locus. When males more negatively effect fitness within the family, a male heterogametic (XY male) sex determining system evolves, whereas when females more negatively effect fitness in the family, a female heterogametic (ZW female) system evolves. Individuals with the dominant sex allele are one sex, and the opposite sex is determined by maternally-expressed genes in individuals without the dominant sex allele. Results therefore suggest that maternal-zygotic conflict could play a role in the early evolution of chromosomal sex determining systems. Predictions are made concerning the patterns of expression of maternal and zygotic sex determining genes expected to result from conflict over sex determination. PMID- 11932769 TI - Pre- and post-insemination episodes of sexual selection in the fowl, Gallus g. domesticus. AB - Although much attention has been recently directed to sexual selection arising after insemination from sexual promiscuity, little is known about the mechanisms determining reproductive success after insemination, and the way these mechanisms interact with each other and with selective mechanisms occurring before insemination: mate choice and mate acquisition. Here, we briefly review the findings of an on-going study investigating the mechanisms generating variation in reproductive success at both a pre- and a post-insemination stage in the domestic fowl. Female preference consistently favours socially dominant males before and after insemination. However, although social status mediates the number of sperm that a male inseminates into a female, dominant males may inseminate sperm of lower fertilising quality than their subordinates. We argue that mitochondrial genes may contribute to determine sperm quality, and speculate that the maternal control of mitochondrial genes may prevent sexual selection from operating on males, thus explaining both the lack of a positive correlation between social dominance and sperm quality and the maintenance of variation in male quality in the fowl. PMID- 11932770 TI - Sex ratios. AB - Sex ratio theory attempts to explain variation at all levels (species, population, individual, brood) in the proportion of offspring that are male (the sex ratio). In many cases this work has been extremely successful, providing qualitative and even quantitative explanations of sex ratio variation. However, this is not always the situation, and one of the greatest remaining problems is explaining broad taxonomic patterns. Specifically, why do different organisms show so much variation in the amount and precision with which they adjust their offspring sex ratios? PMID- 11932771 TI - Origins of the machinery of recombination and sex. AB - Mutation plays the primary role in evolution that Weismann mistakenly attributed to sex. Homologous recombination, as in sex, is important for population genetics -shuffling of minor variants, but relatively insignificant for large-scale evolution. Major evolutionary innovations depend much more on illegitimate recombination, which makes novel genes by gene duplication and by gene chimaerisation--essentially mutational forces. The machinery of recombination and sex evolved in two distinct bouts of quantum evolution separated by nearly 3 Gy of stasis; I discuss their nature and causes. The dominant selective force in the evolution of recombination and sex has been selection for replicational fidelity and viability; without the recombination machinery, accurate reproduction, stasis, resistance to radical deleterious evolutionary change and preservation of evolutionary innovations would be impossible. Recombination proteins betray in their phylogeny and domain structure a key role for gene duplication and chimaerisation in their own origin. They arose about 3.8 Gy ago to enable faithful replication and segregation of the first circular DNA genomes in precellular ancestors of Gram-negative eubacteria. Then they were recruited and modified by selfish genetic parasites (viruses; transposons) to help them spread from host to host. Bacteria differ fundamentally from eukaryotes in that gene transfer between cells, whether incidental to their absorptive feeding on DNA and virus infection or directly by plasmids, involves only genomic fragments. This was radically changed by the neomuran revolution about 850 million years ago when a posibacterium evolved into the thermophilic cenancestor of eukaryotes and archaebacteria (jointly called neomurans), radically modifying or substituting its DNA-handling enzymes (those responsible for transcription as well as for replication, repair and recombination) as a coadaptive consequence of the origin of core histones to stabilise its chromosome. Substitution of glycoprotein for peptidoglycan walls in the neomuran ancestor and the evolution of an endoskeleton and endomembrane system in eukaryotes alone required the origin of nuclei, mitosis and novel cell cycle controls and enabled them to evolve cell fusion and thereby the combination of whole genomes from different cells. Meiosis evolved because of resulting selection for periodic ploidy reduction, with incidental consequences for intrapopulation genetic exchange. Little modification was needed to recombination enzymes or to the ancient bacterial catalysts of homology search by spontaneous base pairing to mediate chromosome pairing. The key innovation was the origin of meiotic cohesins delaying centromere splitting to allow two successive divisions before reversion to vegetative growth and replication, necessarily yielding two-step meiosis. Also significant was the evolution of synaptonemal complexes to stabilise bivalents and of monopolins to orient sister centromeres to one spindle pole. The primary significance of sex was not to promote evolutionary change but to limit it by facilitating ploidy cycles to balance the conflicting selective forces acting on rapidly growing phagotrophic protozoa and starved dormant cysts subject to radiation and other damage. PMID- 11932772 TI - Mate recognition in fungi. AB - The ascomycete and basidiomycete fungi have contributed much to our understanding of eukaryotic cell biology. The study of mate recognition, in particular, has provided detailed understanding of cell signalling pathways and cell-specific gene transcription. Sexual dimorphism has little relevance to mating in these organisms, indeed specialised cells for mating are found only in filamentous ascomycetes and even here, a single individual produces both male and female structures. None the less, most species have genetic barriers to prevent selfing. The genes that determine self-incompatibility divide populations into different mating types, and only individuals with different mating types can engage in sexual reproduction. Ascomycetes have just two mating types, but basidiomycetes may have several thousands. Despite apparent differences in the biology and numbers of mating types in these fungi, it is becoming increasingly apparent that many components of their mating pathways are highly conserved. PMID- 11932773 TI - Sperm competition and the maintenance of polymorphism. AB - Sperm competition may occur whenever sperm from more than one male are present in the reproductive tract of the female. Studies of field-caught Drosophila reveal that a substantial fraction (80%) of females clearly have sperm from more than one male, and the figure is probably higher because only a small number of progeny are typically surveyed, so a strong skew in paternity can make multiply mated females appear as singly mated unless appropriate models are applied. Examination of genetic variation in aspects of sperm competition has revealed some striking patterns, particularly in the implications for the maintenance of polymorphism. The magnitude of variation in sperm competitive ability is as great as that for other fitness components, and the males with the strongest displacement also appear to be the ones with the greatest positive effect on fertility. Why then does not the most competitive allele simply go to fixation? Such synergistic pleiotropy makes the polymorphism even more unexpected. Examination of patterns of competitive success of pairs of male genotypes, and of female-male interactions, demonstrate clearly that the outcome of sperm competition is not a simple property of each male. That is, sperm competitive ability of male genotypes cannot simply be ranked from best to worst. Rather, the outcome of each competitive bout depends on the particular pair of males. These results have intriguing implications for the molecular biology of genes involved in the determination of sperm competitive success, and on the opportunity for maintenance of polymorphism in those genes. PMID- 11932774 TI - Sexual interference of the floral kind. AB - Floral hermaphroditism results in conflicts and compromise in the parental roles of plants during pollination and mating. A potential cost of hermaphroditism is sexual interference between maternal and paternal functions resulting in gamete wastage and reduced fitness. Sexual interference may or may not be associated with self-pollination. In cases where self-pollination occurs, ovule or pollen discounting may reduce mating opportunities. Here I describe forms of sexual interference in flowering plants, distinguishing whether physical or biochemical interactions are involved and whether fitness costs associated with gamete wastage arise from intra-floral versus inter-floral processes. I review the limited experimental evidence for interference between sex functions and evaluate the hypothesis that some floral adaptations usually interpreted as anti-selfing mechanisms may serve an alternative function in reducing mating costs arising from this form of sexual conflict. PMID- 11932775 TI - Ischemic preconditioning prevents postischemic arteriolar, capillary, and postcapillary venular dysfunction: signaling pathways mediating the adaptive metamorphosis to a protected phenotype in preconditioned endothelium. AB - Prolonged ischemia followed by reperfusion (I/R) results in impaired endothelial cell function in all segments of the microvasculature. Moreover, endothelial dysfunction plays a major role in the genesis of the reperfusion component of total tissue injury in I/R. Thus, preservation of endothelial function is an important therapeutic goal for ameliorating injury in tissues subjected to I/R. An accumulating body of evidence indicates that both microvascular endothelium and parenchymal cells can be rendered resistant to the pathological effects of I/R by antecedent exposure to brief periods of ischemia, a phenomenon referred to as ischemic preconditioning (IPC). Although the mechanisms underlying the microvascular effects of preconditioning have been far less extensively studied, work conducted to date indicates that there are fundamental differences in the signaling pathways that underlie the adaptive transformation to a protected or defensive phenotype in the endothelium compared to those that contribute to the development of a preconditioned state in parenchymal cells. Thus, the purposes of this review are to summarize our current understanding of the mechanisms whereby IPC induces the adaptive transformation to a protected or defensive phenotype in parenchymal cells and to compare and contrast this with the signaling pathways that invoke a preconditioned state in arteriolar, capillary, and venular endothelium. In addition, we highlight understudied areas with regard to microvascular protection afforded by antecedent ischemia in the hopes that this will stimulate investigation of the underlying mechanisms. Understanding these signaling pathways may provide a mechanistic rationale for the development of novel treatment interventions that target both the microcirculatory and parenchymal sequelae to I/R, thereby maximizing the therapeutic potential of the protected phenotypes produced by pharmacological preconditioning. PMID- 11932776 TI - Decreased arteriolar sensitivity to shear stress in adult rats is reversed by chronic exercise activity. AB - OBJECTIVE: We tested the hypothesis that the decline in endothelium-dependent arteriolar dilation in adult rats is reversed by chronic exercise activity. METHODS: Rats were divided into young (8-10 weeks)-sedentary (SED(Y)), adult (29 32 weeks)-sedentary (SED(A)), and adult-exercised (EX(A), treadmill exercise for 18-20 weeks) groups. Responses of isolated arterioles ( approximately 50 microm at 80 mm Hg) of gracilis muscle were assessed to increases in perfusate flow and vasoactive agents. RESULTS: With no differences in basal tone, maximal flow induced dilations were not different between arterioles of SED(Y) and SED(A) rats (71 +/- 2 and 72 +/- 2% of passive diameter, respectively), yet the sensitivity of arterioles to shear stress (WSS(50)) was significantly less in SED(A) than in SED(Y) rats (35 +/- 4 vs. 23 +/- 3 dyne/cm(2), respectively). In vessels of EX(A) rats, maximal flow-induced dilation was significantly augmented (88 +/- 2% of passive diameter) and WSS(50) (15 +/- 1 dyn/cm(2)) was significantly reduced. Dilation to acetylcholine was enhanced in arterioles of EX(A), whereas dilation to sodium nitroprusside was not different in vessels of the three groups. CONCLUSION: Chronic exercise activity reverses age related reduction in sensitivity of arterioles to increases in wall shear stress. PMID- 11932777 TI - T-lymphocytes modulate the microvascular and inflammatory responses to intestinal ischemia-reperfusion. AB - OBJECTIVE: The overall objective of this study was to define the contribution of T-lymphocytes to the microvascular and inflammatory responses of the intestine to ischemia/reperfusion (I/R). METHODS: The superior mesenteric artery of wild-type (WT) and SCID mice was occluded for 45 minutes, followed by 30 minutes or 6 hours of reperfusion. Intravital fluorescence microscopy was used to monitor the extravasation of FITC-labeled albumin or the adhesion of carboxy-fluorescein diacetate succinimidyl ester (CFSE)-labeled T-lymphocytes in mucosal venules of the postischemic intestine. Tissue myeloperoxidase (MPO) was used to monitor neutrophil accumulation in the intestine of WT and SCID mice. RESULTS: Although the number of adherent T-cells was not increased above baseline at 1 hour after reperfusion, significant T-cell adhesion (both CD4(+) and CD8(+)) was noted at 6 hours of reperfusion. The latter response was prevented by pretreatment with a blocking antibody directed against MAdCAM-1, but not ICAM-1 or VCAM-1. A significant increase in MAdCAM-1 expression was noted in both lymphoid (Peyer's patch) and nonlymphoid regions of the postischemic small bowel. The early (30 minutes after reperfusion) albumin extravasation elicited by gut I/R in WT mice was reduced in SCID mice. Reconstitution of SCID mice with T-lymphocytes restored the albumin leakage response to WT levels. The increased intestinal MPO caused by I/R (6 hours of reperfusion) in WT mice was attenuated in SCID mice; with reconstitution of SCID mice with T-cells the MPO response was restored. CONCLUSIONS: These findings indicate that intestinal I/R is associated with the recruitment of CD4+ and CD8+ T-cells, which is mediated by endothelial MAdCAM-1. T-cells seem to modulate the recruitment of neutrophils that occurs hours after reperfusion as well as the increased albumin extravasation that occurs within minutes after reperfusion. PMID- 11932778 TI - Hemodynamic stresses and structural remodeling of anastomosing arteriolar networks: design principles of collateral arterioles. AB - OBJECTIVE: To investigate the potential influence of hemodynamic stresses on the development of the arcade arteriole (AA) network during normal maturation. METHODS: AA network data were collected from ink-filled Wistar-Kyoto rat gracilis muscles and used to construct hemodynamic computational models of the AA network at 7 (WKY(7)) and 13 (WKY(13)) weeks of age. RESULTS: Mean coefficients of variation for pressure, circumferential wall stress, and wall shear stress were 0.13, 0.12, and 0.48, respectively. Wall shear rate variability across bifurcations generated deviations in mean energy cost that were 9-30% above theoretical minimum, with many bifurcations exhibiting substantially higher energy costs. With the exception of the lowest pressure AA segments, the monotonic relationship between wall shear stress and pressure in the AAs was nearly identical from 7 to 13 weeks of age. CONCLUSIONS: Low coefficients of variation for computed AA pressures indicate that an even pressure head is maintained over the muscle during remodeling of the AA network. The anastomotic structure of the network creates high shear rate variability that, in turn, creates high-energy costs in some regions of the network. The results are consistent with the hypothesis that, during development, the maintenance of mean circumferential wall stress and the pressure-shear stress relationship are operative design principles for collateral arteriole development. PMID- 11932779 TI - Effect of urate on nitric oxide microcirculatory response in the rat tail to body heating. AB - OBJECTIVE: The objective of this study was to determine if nitric oxide (NO) mediated cutaneous vasodilation depends on urate concentrations. METHODS: Capillary blood cell velocities (BCV) in the tail capillary vessels of anesthetized, heat-stressed Sprague-Dawley rats were measured by the videomicroscopy method, and the femoral mean arterial pressure (MAP) was recorded on a Grass polygraph. The effects of urate or urate and NO production on the above parameters were determined by using specific inhibitors of these compounds, for example, allopurinol (All) or All or N-nitro-L-arginine (NNA), respectively. Urate concentrations were determined by an enzymatic, colorimetric method, and total nitrite levels were measured fluorometrically. RESULTS: During heat stress, All or All and NNA significantly decreased plasma urate or urate and total nitrite concentrations, respectively, and the response of vascular conductance (estimated as a ratio of BCV to MAP). Urate administration, after All and NNA pretreatment, restored plasma urate and total nitrite concentrations and increased BCV/MAP. CONCLUSIONS: Urate at physiological concentration helps to maintain vascular conductance of cutaneous capillaries in the rat tail during body heating. The effect of urate on vascular conductance may be mediated by NO. PMID- 11932780 TI - Expression and regulation of the PD-L1 immunoinhibitory molecule on microvascular endothelial cells. AB - OBJECTIVE: To evaluate the expression and regulation of a novel B7-like protein, PD-L1, the ligand for the immunoinhibitory receptor PD-1 expressed on activated T cells, on microvascular endothelial cells (ECs) METHODS: PD-L1 expression on ECs in vitro and in vivo was quantified by using a dual radiolabeled antibody technique after treatment with interferons (IFN) and IL-12, respectively. Changes in the level of PD-L1 mRNA were determined by using RT-PCR. RESULTS: PD-L1 was observed to be present on ECs under basal conditions. Treatment of ECs with IFN alpha, -beta and -gamma, but not LPS, was observed to induce elevations in the mRNA and surface expression of PD-L1 on ECs. By using a dual radiolabeled monoclonal antibody (mAb) technique, PD-L1 expression in various tissues of control and IL-12 challenged wild-type and IFN-gamma-deficient mice was measured. A significant increase in PD-L1 expression was observed in tissues at 24 hours after IL-12-challenge, with peak levels of PD-L1 occurring 72 hours after IL-12 challenge. IL-12 was not effective at inducing PD-L1 expression in tissues of IFN gamma-deficient mice. CONCLUSIONS: These data show the expression of a novel B7 like molecule on murine ECs that is mediated by IFN-alpha, -beta, and -gamma, and suggest a potential pathway by which ECs may modulate T-cell function. PMID- 11932781 TI - Long-term follow-up of patients after endoscopic sphincterotomy for choledocholithiasis, and risk factors for recurrence. AB - BACKGROUND AND STUDY AIMS: There have been growing concerns about the long-term sequelae of endoscopic sphincterotomy (ES). The aims of the study were to evaluate the long-term clinical outcome of ES for choledocholithiasis and to identify the predictors of recurrence. PATIENTS AND METHODS: A total of 529 patients (233 men, 296 women; mean age 63, range 9 - 88) who underwent successful ES for choledocholithiasis were evaluated. Follow-up data were obtained retrospectively. Statistical analyses were carried out for 334 patients who had a follow-up of at least 5 years or had recurrence. RESULTS: Immediate complications occurred in 37 patients (7 %). Follow-up data were available in 458 patients (86.6 %), of whom 280 (61.1 %) were asymptomatic, 127 (27.7 %) died from unrelated causes without recurrence, and 51 (11.1 %) had biliary symptoms and/or choledocholithiasis recurrence. Most recurrences (65 %) occurred more than 2 years following ES and were observed on multiple occasions in 13 patients (2.8 %). A bile duct diameter of 22 mm or greater was found to predict recurrence. Of 190 patients with an intact gallbladder, 11 (5.8 %) developed acute cholecystitis necessitating emergency cholecystectomy; all of these had previously documented gallstones. CONCLUSIONS: Endoscopic sphincterotomy for choledocholithiasis is found to be safe at long-term follow-up. A dilated bile duct (> or = 22 mm) is a marker for patients at increased risk of recurrence of symptoms and/or choledocholithiasis. PMID- 11932782 TI - Sphincter of Oddi dysfunction: pancreaticobiliary sphincterotomy with pancreatic stent placement has a lower rate of pancreatitis than biliary sphincterotomy alone. AB - BACKGROUND AND STUDY AIMS: Patients with suspected or documented sphincter of Oddi dysfunction (SOD) who undergo standard biliary sphincterotomy have high rates of post-procedure pancreatitis. Approximately 75% of such patients have elevated basal pressures of the pancreatic sphincter. Biliary sphincterotomy (BES) on its own leaves the pancreatic sphincter unablated and may cause transient edema which aggravates the increase in pancreatic sphincter pressure. Combined pancreaticobiliary therapy (PBR), using pancreatic stenting in addition to sphincterotomy may therefore be safer. PATIENTS AND METHODS: The endoscopic retrograde cholangiopancreatography (ERCP) database was queried for patients with successful double-duct sphincter of Oddi manometry (SOM) who underwent BES alone or PBR between 1994 and 1997. The endoscopist had decided on the technique to be used. From 1995 to 1997 there was a general trend to do PBR. Pancreatitis was defined according to established criteria. RESULTS: The post-ERCP pancreatitis rate among all 436 SOD patients was 19.7%, while 256 patients with normal SOM results had a pancreatitis rate of 12.9%. The use of combined PBR was associated with a lower frequency of pancreatitis compared with BES alone (needle-knife over pancreatic duct stent, 14/131 patients, 10.7 %; pull-type pancreaticobiliary sphincterotomy plus pancreatic stent, 15/78 patients, 19.2%; BES alone, 52/184 patients, 28.3%). Episodes of moderate and severe pancreatitis were seen more frequently in the BES group. CONCLUSION: In SOD patients, post-ERCP pancreatitis rates remain high, but have improved with the addition of combined pancreaticobiliary sphincter therapy. PMID- 11932783 TI - Post-ERCP pancreatitis and hyperamylasemia: patient-related and operative risk factors. AB - BACKGROUND AND STUDY AIMS: Previous studies on the development of post-endoscopic retrograde cholangiopancreatography (post-ERCP) hyperamylasemia and pancreatitis have focused on different risk factors, either procedure- or patient-related, and also prognostic factors, such as amylase levels together with the occurrence of pancreatic-type pain, which might be implicated. The aim of this study is to identify possible predictive risk factors and also prognostic factors in order to better organize the treatment strategy. PATIENTS AND METHODS: During a 5-year period, 556 ERCP procedures were performed by the same operator. Of these, 43 procedures were excluded from the study. Data recorded from the procedures included both patient characteristics and procedure details. Patients were evaluated for pancreatic-type pain at 4 and 24 h after the procedure, and serum amylase levels were determined at 2 and 24 h after the procedure. Pancreatitis was diagnosed on the basis of both clinical and laboratory examination, while patients with hyperamylasemia were in a normal clinical condition but had high serum amylase levels. RESULTS: Post-ERCP pancreatitis occurred after 17 procedures (3.3%) and hyperamylasemia after 85 procedures (16.5%). Patients who developed pancreatitis all had pancreatic-type pain together with amylase levels higher than 4-5 times the upper normal limit at 24 h after ERCP. There was a significant association between serum amylase levels and post-procedure pancreatic-type pain both at 4 h and 24 h after ERCP (P = 0.006). Age less than 50 years, history of relapsing pancreatitis, pancreatic duct opacification, and difficulty in obtaining bile duct cannulation all proved to be significant predictive risk factors for the development of either hyperamylasemia or pancreatitis. In contrast, additional procedures and sphincterotomy seemed to reduce the likelihood of both complications. CONCLUSIONS: Serum amylase levels higher than 4 - 5 times the normal upper limit together with the existence of pancreatic-type pain, at 24 h after ERCP, strongly suggest the occurrence of pancreatitis. When any of the predictive risk factors coexist during a procedure it would be better either to abandon the procedure and try again later or, if the situation is urgent, to have earlier recourse to an alternative, such as precut or needle-knife papillotomy. PMID- 11932784 TI - ERCP-related perforations: risk factors and management. AB - BACKGROUND AND STUDY AIMS: Perforations during endoscopic retrograde cholangiopancreatography (ERCP) are rare, and the management of these perforations is variable, with some patients requiring immediate surgery and others only conservative management. We reviewed all ERCP-related perforations at our institution to determine: a) their incidence; b) clinical outcomes; c) which management approaches gave the best results; and d) which factors predict a perforation. PATIENTS AND METHODS: All patients who underwent ERCP and suffered perforation were reviewed. To compare the length of hospital stay of the perforation group with that of patients suffering a different complication, patients who developed post-ERCP pancreatitis were also reviewed. To evaluate predictors of ERCP-related perforations, three groups were compared: group 1 (n = 49), normal ERCP/no complications; group 2 (n = 52), ERCP complicated by pancreatitis; and group 3 (n = 33), ERCP with perforation. RESULTS: Of 33 patients with confirmed ERCP-related perforations, only seven patients required surgical intervention. The overall length of hospital stay (6.5 +/- 3.5 days) was significantly longer (P = 0.003) than that of a random group of patients with the complication of post-ERCP pancreatitis (4.7 +/- 2.6 days). According to univariate analysis, risk factors included: sphincterotomy (odds ratio [OR] 9.0, 95 % confidence interval [CI] 3.2 - 28.1); sphincter of Oddi dysfunction (OR 3.8, 95 % CI 1.4 - 11.0); and dilated common bile duct (OR 4.07, 95 % CI 1.63 - 10.18, P = 0.003). In the multivariate logistic regression analysis, additional predictive factors included the duration of procedure (OR 1.021, 95 % CI 1.006 - 1.036), and biliary stricture dilation (OR 7.2, 95 % CI 1.84 - 28.11). CONCLUSIONS: (i) The incidence of ERCP-related perforations is very low (0.35 %). (ii) Esophageal, gastric and duodenal perforations usually require surgery, but sphincterotomy- and guide wire-related perforations rarely do so. (iii) Factors which carry increased risk of an ERCP-related perforation include suspected sphincter of Oddi dysfunction, greater age, a dilated bile duct, sphincterotomy, and longer duration of the procedure. PMID- 11932785 TI - Accuracy of linear endoscopic ultrasonography in the evaluation of patients with suspected common bile duct stones. AB - BACKGROUND AND STUDY AIMS: Endoscopic ultrasonography (EUS) with radial scanning is an efficient diagnostic tool where there is suspicion of common bile duct (CBD) stones. Little is known about the use of linear EUS in this condition. The aim of this study was to evaluate the diagnostic efficiency of linear EUS in a large group of patients suspected to have bile duct stones, using endoscopic retrograde cholangiopancreatography (ERCP) with endoscopic sphincterotomy and exploration of the CBD using a Dormia basket, or surgical choledochotomy with choledochoscopy, as diagnostic "gold standards." PATIENTS AND METHODS: 134 patients with clinical suspicion of CBD stones were included in the study and prospectively evaluated, using EUS, and ERCP with endoscopic sphincterotomy (127 patients), or choledochotomy with choledochoscopy where ERCP was unsuccessful (seven patients). EUS was done before ERCP using an echo endoscope (Pentax FG 32 UA; 5 - 7.5 MHz) and Hitachi EUB 405 ultrasound machine. ERCP was done using the TFJ 100 or TJ 20 Olympus duodenoscope. ERCP was carried out within a mean of 2 days after EUS. The longest time between EUS and ERCP was 3 days. The examiners were blinded to the results of the other method used. RESULTS: CBD stones were found in 91 (68 %) patients at ERCP with ES or at surgery. The correct diagnosis was established by EUS in 85 patients. The remaining 43 patients without CBD stones were correctly diagnosed in 41 cases by means of EUS, giving an accuracy of 94 %, sensitivity of 93 %, specificity of 93 %, a positive predictive value of 98 %, a negative predictive value of 87 %, and a Youden's index of 89 %. CONCLUSIONS: Linear EUS is a fairly reliable method for the evaluation of patients with high suspicion for CBD stones. The usefulness of linear EUS in the evaluation of patients with low or moderate suspicion for CBD stones warrants further study. PMID- 11932787 TI - The clinical significance of colonoscopy in hemorrhagic colitis due to enterohemorrhagic Escherichia coli O157:H7 infection. AB - BACKGROUND AND STUDY AIMS: Although hemorrhagic colitis due to enterohemorrhagic Escherichia coli O157:H7 (O157) infection has recently attracted increasing attention as an important enteric infection, the colonoscopic findings associated with this disease have not been sufficiently characterized. The aim of this study is to characterize the colonoscopic features of hemorrhagic colitis due to O157 infection. PATIENTS AND METHODS: The colonoscopic findings in ten patients with hemorrhagic colitis due to O157 infection were retrospectively reviewed. To assess the severity of inflammation in each part of the large intestine, colonoscopic findings were categorized into four grades: grade 1, intact mucosa; grade 2, sporadic erythema and erosion; grade 3, mostly diffuse inflammation; and grade 4, diffuse, severe inflammation. RESULTS: Eight out of ten patients had grade 4 findings in the cecum and ascending colon, grade 3 in the transverse colon and descending colon, and grade 2 in the sigmoid colon. Two of these eight patients also had grade 4 inflammation in the proximal transverse colon. Five of these eight patients revealed longitudinal ulcer-like lesions in the transverse colon and/or descending colon. The remaining two patients had grade 3 findings in the cecum to the descending colon and grade 2 findings in the sigmoid colon. All patients exhibited grade 1 finding in the terminal ileum and the rectum. Based on these colonoscopic findings, the ten patients were divided into the typical group (eight patients) and the mild-type group (two patients). CONCLUSIONS: The characteristic colonoscopic findings in most patients with hemorrhagic colitis due to O157 infection were as follows: 1) severe inflammation, including primarily marked edema and facile hemorrhage, and 2) inflammation predominating at the right-side colon; and 3) frequent appearance of longitudinal ulcer-like lesions. PMID- 11932786 TI - What is the utility of selected clinical and endoscopic parameters in predicting the risk of death after caustic ingestion? AB - BACKGROUND AND STUDY AIMS: Despite the increasing use of early esophagogastroduodenoscopy, the prognostic evaluation and triage of patients who have ingested caustic material is challenging. We evaluated the usefulness of selected clinical and endoscopic parameters in predicting the risk of death after ingestion of caustic substances. PATIENTS AND METHODS: Clinical and endoscopic parameters were obtained from the records of all the patients admitted to our endoscopy unit because of ingestion of caustic material between 1 March 1982 and 30 June 1999. Parameters significantly associated with the risk of death by univariate analysis were entered into a multivariate logistic model. The independent predictors of death by multivariate analysis were used to build a risk score system. RESULTS: Out of 210 patients, 13 underwent emergency surgery (6.2 %) and 25 died (11.9 %). Multivariate analysis identified the following as independent predictors of death: age (10-year intervals; odds ratio [OR] 2.4; 95 % confidence interval 1.4 - 4.1), ingestion of strong acids (OR 7.9; 1.8 - 35.3), white blood cell count at admission > or = 20 000 units/mm3 (OR 6.0; 1.3-28), deep gastric ulcers (OR 9.7; 1.4 - 66.8), and gastric necrosis (OR 20.9; 4.7 - 91.8). The values of the risk score system devised from the results of the multivariate analysis ranged from 1 to 16. No patient scoring < 10 points died and just one of the patients scoring > 14 points survived. CONCLUSION: Age, ingestion of a strong acid, leucocytosis, deep gastric ulcers, and gastric necrosis are predictive of death after caustic ingestion. A risk score system including these predictors may be useful in prognostic evaluation. PMID- 11932788 TI - Latent adenocarcinoma of the colon discovered during or after laparoscopic cholecystectomy. AB - BACKGROUND: The wide acceptance of laparoscopic cholecystectomy has resulted in the performance of increased numbers of cholecystectomies, hence increasing the number of patients discovered with concomitant malignancy or other pathological states. PATIENTS AND METHODS: A total of 3751 patients who underwent laparoscopic cholecystectomy between January 1995 and December 2000 were included in this study. Nine cases of coexisting malignant colonic neoplasm were discovered. All of these patients but one underwent therapeutic resection. RESULTS: The survival rate of these nine patients was the same as that of 62 consecutive colonic cancer patients who were treated in our department over the same period. The rate of postoperative complications in the study group was higher than in the control group (21 % vs. 9.5 %). The hospital stay in these patients was also greater (17 days vs. 10 days). CONCLUSION: The rate of incidental discovery of colorectal carcinomas during laparoscopic cholecystectomy in our department was 0.24 %. These patients had higher complication rates and a longer hospital stay. PMID- 11932789 TI - Laparoscopic features of primary biliary cirrhosis in AMA-positive and AMA negative patients. AB - BACKGROUND AND STUDY AIMS: Antimitochondrial antibody (AMA)-negative primary biliary cirrhosis (PBC) has been difficult to diagnose. Laparoscopic features of AMA-negative PBC were evaluated in comparison with those of AMA-positive PBC and autoimmune hepatitis. PATIENTS AND METHODS: 71 patients who fulfilled the diagnostic criteria for PBC were enrolled in the study; 48 were AMA-positive and 23 were AMA-negative. As a disease control, 46 autoimmune hepatitis patients were included. Both the frequency and specificity of each laparoscopic finding were evaluated. A laparoscopic scoring system was introduced, which used, common and uncommon laparoscopic findings, and was evaluated for the diagnosis of AMA negative PBC. RESULTS: The characteristic laparoscopic findings for AMA-positive PBC were yellowish-white marking (92 %), dark-brown discoloration (73 %), gentle undulation (67 %), reddish patch (38 %), and yellowish-white nodules (32 %). On the other hand, laparoscopic findings such as trench-like depression, reddish markings, and wide and small depressions were uncommon in PBC compared with autoimmune hepatitis. The frequencies of characteristic and uncommon laparoscopic findings did not differ statistically between AMA-positive and AMA-negative PBC, but were different between AMA-positive or AMA-negative PBC and autoimmune hepatitis. Scores based on common and uncommon laparoscopic findings were 5.5 +/- 1.5 (mean +/- SD) in AMA-positive PBC, 5.6 +/- 2.0 in AMA-negative PBC, and - 0.30 +/- 0.5 in autoimmune hepatitis. CONCLUSION: The laparoscopic findings in AMA-negative PBC did not differ from those of AMA-positive PBC. A laparoscopic scoring system may be helpful in the diagnosis of AMA-negative PBC. PMID- 11932791 TI - Post-ERCP pancreatitis: new momentum. PMID- 11932790 TI - Antibiotic prophylaxis of infective endocarditis during digestive endoscopy: over and underuse in Switzerland despite professed adherence to guidelines. AB - BACKGROUND AND STUDY AIMS: Guidelines for antibiotic prophylaxis of infective endocarditis associated with gastrointestinal endoscopy are not uniform. Though based on weak scientific evidence, they do represent an attempt to unify the management of antibiotic prophylaxis. We investigated whether physicians who profess to adhere to these guidelines actually do apply them correctly. MATERIALS AND METHODS: A questionnaire was sent to all 195 Swiss gastroenterologists asking whether they adhered to antibiotic prophylaxis guidelines and under what conditions did they apply antibiotic prophylaxis in gastrointestinal endoscopy. We analysed whether the Swiss gastroenterologists who claim to adhere to these guidelines actually do so in practice. RESULTS: The response rate to our questionnaire was 95%. Of the Swiss gastroenterologists, 60% correctly employ antibiotic prophylaxis guidelines in gastroscopy, as do 47% in colonoscopy. In therapeutic endoscopy, the percentage of correct antibiotic prophylaxis use depends upon the type of intervention and varies between 21 and 58%. Overuse of antibiotic prophylaxis is at least twice as frequent in colonoscopy and therapeutic endoscopy than in gastroscopy. CONCLUSIONS: Guidelines for antibiotic prophylaxis are not well applied and do not lead to uniform management despite a high degree of professed adherence to them. This phenomenon can be explained by "cognitive dissonance" and "reactance." We conclude that guidelines based on unconvincing data may even prove harmful. PMID- 11932792 TI - Chromoscopy of intestinal metaplasia in Barrett's esophagus. PMID- 11932793 TI - Endoscopic diagnosis of a right hepatic artery pseudoaneurysm. AB - Hepatic artery aneurysms are rare occurrences with diverse etiologies. We present a case of a right hepatic artery aneurysm, which was diagnosed at endoscopic retrograde cholangiography (ERC) and treated angiographically. This is the first report where ERC has been critical in delineating the aneurysmal cavity, suggesting the diagnosis and prompting emergency intervention. PMID- 11932794 TI - Endosonography-guided celiac plexus neurolysis in the treatment of pain secondary to acute intermittent porphyria. AB - Acute intermittent porphyria is a metabolic error transmitted as an autosomal dominant disorder with incomplete penetrance. Its clinical picture includes intermittent abdominal pain, nausea, vomiting, and diarrhea, with or without neurological changes. We report the case of a young woman whose pain attacks were controlled with high-dose opiates, in whom we decided to perform endosonography guided celiac plexus neurolysis (EUS-CPN). This is the first reported attempt with this new treatment option. There was significant clinical and nutritional improvement after treatment. PMID- 11932795 TI - The possible involvement of micro-organisms other than Helicobacter pylori in the development of rectal MALT lymphoma in H. pylori-negative patients. AB - It remains unclear whether lymphoma of the mucosa-associated lymphoid tissue (MALT) in the extragastric organs is related to Helicobacter pylori infection or not. This report describes three patients with rectal MALT lymphoma negative for H. pylori infection, all of whom showed disease regression after being treated with antibiotics. One patient had MALT lymphoma in both the descending colon and the rectum; the other two patients had rectal disease only. None of the patients had chronic gastritis which was detectable either endoscopically or histologically and H. pylori infection was completely ruled out by various methods, including a urease breath test. These patients received antibiotic therapy. In all the patients, regression of MALT lymphoma was observed endoscopically and histologically, and polymerase chain reaction revealed that a previously observed rearranged band of immunoglobulin heavy chain had also disappeared after antibiotic treatment. These cases therefore suggest involvement of micro-organisms other than H. pylori in the development of rectal MALT lymphoma. PMID- 11932797 TI - Use of an overtube for enteroscopy: depth of insertion. PMID- 11932798 TI - From laparoscopic-assisted to scarless appendicectomy: continuing advances. PMID- 11932799 TI - An unusual endoscopic feature in the duodenum of a young girl with intraluminal duodenal diverticulum. PMID- 11932800 TI - Duodenal perforation secondary to placement of a biliary endoprosthesis diagnosed by multislice computed tomography. PMID- 11932801 TI - Generalized and mucosal melanosis associated with ultra-late recurrence of malignant melanoma. PMID- 11932802 TI - Impacted foreign body at the pharyngoesophageal junction: an innovative management. PMID- 11932804 TI - [Clinical monthpaper 2002 - Editorial]. PMID- 11932803 TI - Upper gastrointestinal bleeding from leukemic gastric implants. PMID- 11932805 TI - [Plastics on the eye - plastics in the eye. Human-optic materials]. AB - BACKGROUND: Materials of identical or similiar chemical classes are used for contact optics as well as for implantable intraocular devices. All of them derive from the chemistry of industrial product developments. PATIENTS AND METHODS: Materials used in and on the eye should behave indifferent. They should not provoke any reaction in the eye and not themselves be altered by the biological surroundings. For materials used in the eye an especially high purity of the polymers utilized as well as a long term light stability is to be demanded. Optical properties like transparency have to be stable over extended time periods under normal light influence. The long time stability of UV-absorbing additives in the materials used has to be verified. RESULTS: All materials used for IOL's - with the exception of the homopolymer of methylmethacrylate - consist of polymer mixtures forming socalled interpenetrating networks which appear as an uniform material. For the manufacturing of intraocular lense materials standardized or unanimous producer prescriptions are not existing. Based on the different polymer formulations different aging and fatigue properties are the outcome. In addition only inadequate knowledge about the long term light stability of the incorporated UV blockers is submitted. CONCLUSIONS: Polymeric materials used for IOL's should be subjected in addition to the test methods listed in EN-ISO 11 979/5 to chemical, polymeranalytic and mechanical substance examinations. As a standard in such examinations a polymethylmethacrylate, free of methylacrylate is proposed. Examinations of each lot should be compulsory. Identification of all ingredients of the intraocular materials should be prescribed and labeled in line with the revelations of common medical drugs prescriptions. The relevations commit the medical doctor to inform his patient about possible side effects of the intraocular implants. PMID- 11932806 TI - [Excimer laser subepithelial ablation (ELSA) or laser epithelial keratomileusis (LASEK) - a new kerato-refractive procedure for myopia. Surgical technique and first clinical results on 24 eyes and 3 months follow-up]. AB - BACKGROUND: Laser epithelial keratomileusis (LASEK) is a new keratorefractive surgical procedure to treat myopia and possibly other refractive errors, which combines the advantages while eliminating the disadvantages of both PRK (pain, corneal haze) and LASIK (flap complications, interface complications). Our results with LASEK are very promising and because this technique is currently not very popular in Germany we here with introduce the surgical technique and our first clinical results. PATIENTS AND METHODS: We treated 24 eyes (18 patients) by LASEK. The preoperative myopia was between - 1.50 and - 6.00 D with an astigmatism less than 1.00 D. Surgery starts with an incision in the epithelium by using a special microtrephine. After the application of 20 % ethanol for 20 seconds a hinged epithelial flap is created and the laser ablation performed with a Keracor 117 excimer laser. Finally, the epithelial flap is repositioned and secured using a bandage contact lens and artificial tears for 3 days. RESULTS: In all eyes the creation of the epithelial flap and its repositioning presented no problems. None of the eyes showed any postoperative epithelial instability. However, one eye lost the contact lens and the epithelial flap during the first postoperative day. Postoperatively, none of the patients complained of significant pain; however, the contact lens caused some foreign body sensation. After 3 months all eyes were within +/- 1.0 D and 20/24 eyes (84 %) within +/- 0.50 D of emmetropia (spherical equivalent). None of the eyes showed significant haze. CONCLUSIONS: Although, the number of treated eyes is small and the postoperative follow-up is short LASEK seems to be an effective and safe procedure for the treatment of myopia. PMID- 11932807 TI - [Positive influence of color experience on result of color-arrangement-test Roth 28-hue (E) desaturated - a clinical study on 44 patients]. AB - BACKGROUND: The color arrangement test can be applied for early diagnosis of diabetic retinopathy, even small confusion of colors influence clinical diagnosis. It is therefore necessary to be aware of influential factors. METHODS: Forty-four patients with color-experience (VW-Werk Wolfsburg) were included and devided in two groups: group 1: non-smokers without ophthalmological and systemic diseases (n = 27, 42+/-9 years), group 2: smokers without ophthalmological and systemic diseases (n = 17; 43+/-8 years). The control group 3 (n = 30; 42+/-4 years) included non-smokers and the control group 4 (n = 10; 42+/-8 years) smokers, both groups without color-experience, ophthalmological and systemic diseases. Besides the ophthalmological examinations (visual acuity, refraction, intraocular pressure, slit lamp and fundus examination) the color-vision was tested by the color-arrangement-test Roth 28-hue (E) desaturated monocularly under standard conditions: The background used was black cardboard, illuminated by two Osram fluorescent lamps (L36W/12LDL Daylight) providing 2000 lux at the test table. RESULTS: Ophthalmological examination in all subjects was without pathological findings. The mean error score in the non-smokers with color experience (median+/-mean absolute deviation: 48+/-47) was lower than in the non smokers without color experience (72+/-45; Mann-Withney-U-Test: p = 0.02). The mean error score in the smokers with color-experience (60+/-60) was lower than in the smokers without color-experience (156+/-65; p = 0.0014). No statistically significant difference was found between the measurements of the right and left eye (Wilcoxon-Test: group 1: p = 0.89; group 2: p = 0.9; group 3: p = 0.77; group 4: p = 0.87). CONCLUSION: Color experience improves the results in color arrangement-tests like the Roth 28-hue (E) desaturated and should be considered in quantitative evaluation. PMID- 11932808 TI - [Transformation of cystic into diffuse epithelial ingrowth caused by laser puncture. Report on four patients]. AB - BACKGROUND: Epithelial ingrowth to the anterior chamber is a rare event after ocular injury or intraocular surgery. We report on four patients with primary cystic epithelial ingrowth (with primary curative cystic formation when treated with block excision) who underwent Nd:YAG-laser puncture of the cysts elsewhere. This treatment led to a change from a cystic to a diffuse form with a poor prognosis for the eye. PATIENTS: Three women and one man were seen in our outpatient department. Cystic epithelial ingrowth happened after intraocular surgery (1 x goniotomy, 2 x cataract surgery) or perforating injury (1 x). All four patients were treated elsewhere with puncture of the cysts (by Nd:YAG laser). At first presentation in our department a change of cystic to diffuse epithelial ingrowth was noticed. Curative treatment by block excision was no longer possible. Diffuse epithelialisation of the anterior chamber resulted in secondary glaucoma with a poor prognosis. CONCLUSIONS: In our experience puncture of intraocular epithelial cysts may lead to a transformation to diffuse epithelial ingrowth with possible loss of the eye. Surgical opening of the cyst or subtotal excision is therefore contraindicated from our point of view. In case of cystic epithelial ingrowth of less than 5 hours of the limbal circumference, block excision in toto is indicated. PMID- 11932809 TI - [Visual acuity and astigmatism after eccentric penetrating keratoplasty - a retrospective study on 117 patients]. AB - PURPOSE: The purpose of this study was to asses the functional results of eccentric homologous penetrating keratoplasty in respect of diagnosis as well as position and size of the corneal graft. PATIENTS AND METHODS: In this retrospective study, visual acuity and astigmatism of 117 cases of a round eccentric homologous penetrating keratoplasty in 91 eyes of 91 patients were analysed. The PK became necessary because of a corneal ulcer (104), 6 eyes with corneal scars and 7 eyes with ectatic corneal diseases. Using 84 postoperative slides the following parameters were quantified: graft diameter and transparency, distance of the geometric centre of the cornea from the central trephination margin (distance) as well as from the geometric centre of the graft (decentration). Seventeen grafts were classified as eccentric peripheral keratoplasty (mean diameter 5.0 +/- 1.7 mm) with the optical axis through the host cornea and 67 as eccentric central keratoplasty (7.0 +/- 1.3 mm) with the optical axis through the graft. RESULTS: The average follow-up period was 25 months. The astigmatism after 3/12/24 months was 6.7/6.3/5.5 D, but was irregular and not measurable in 74 %/52 %/49 %. The astigmatism increased with increasing decentration (24 months, p = 0.04). After 3/12/24 months, the VA in ulcers was 0.1/0.16/0.16, in scars 0.2/0.6/0.5 and in ectatic diseases 0.3/0.5/0.6. The position of graft and the visual acuity did not correlate significantly. After 24 months, larger the graft size was associated with better visual acuity. With increasing decentration the visual acuity decreased (p = 0.02) and was significantly lower (0.15) in the case of - 2.0 < distance < 2.0 mm than in the case of distance > 2.0 mm (0.4; p = 0.01). CONCLUSION: The diagnosis resulting in eccentric penetrating keratoplasty seems to have a major impact on the long-term functional prognosis of visual acuity. In addition, visual acuity is limited by high and often irregular astigmatism. Our results indicate that an increasing decentration of the graft is not invariably associated with decreasing visual acuity, provided the graft-host junction is not located in direct proximity of the optical axis. PMID- 11932810 TI - [The predictive value of a classification for proliferative diabetic vitreoretinopathy]. AB - BACKGROUND: Kroll's classification of proliferative diabetic vitreoretinopathy (PDVR) defines stage A (vitreoretinal proliferations without retinal detachment), stage B (partially detached retina not involving the macula), stage C (partially detached macula) and stage D (completely detached macula). The purpose of this study was to analyse the prognostic value of Kroll's classification in respect of the postoperative results of vitreoretinal surgery based on our group of patients. PATIENTS AND METHODS: The charts of 563 patients who underwent vitrectomy because of PDVR between 1990 and 1997 were examined retrospectively. Postoperative visual acuity, possible risk factors, frequencies of silicone oil tamponade and revitrectomies were related to the preoperative staging of PDVR. The influence of possible predictive factors on the postoperative visual outcome were evaluated using multivariate logistic regression analysis. RESULTS: After vitreoretinal surgery mean postoperative visual acuity was significantly better in stage A compared to stage C (p < 0.01) or D (p < 0.0001). In 179 out of 563 eyes (31.7 %) revitrectomy (including silicone oil removal) was required and in 51 eyes (9.1 %) more than one revitrectomy was performed. Silicone oil tamponade was used in 22 out of 253 eyes (8.7 %) classified as stage A, in 27 out of 201 eyes (13.4 %) of stage B, in 17 out of 78 eyes (21.8 %) of stage C and in 10 of 31 eyes (32.3 %) of stage D. Postoperative increase of visual acuity of more than 3 lines was significantly less frequent in stage B (p < 0.014), C (p < 0.039) and D (p < 0.001) compared to stage A. CONCLUSION: Kroll's classification for PDVR has a high prognostic value for the postoperative visual outcome and level of surgical risk management. Thus patients with good prognosis can be identified easily, facilitating the decision for surgery. PMID- 11932811 TI - [Cyclotropia and surgical treatment after macular translocation]. AB - PURPOSE: Macular translocation is a new surgical treatment method successfully performed in recent years in cases of age-related macula degeneration. Cyclotropia and associated orthoptic problems may occur after macular translocations. The aim of our study was to present the application and results of various external eye muscle surgery techniques to eliminate cyclodeviation after macular translocation. METHODS: Between January 2001 and April 2001, 15 patients underwent macular translocation. After an average of 47 days counterrotation was applied to these patients to eliminate incyclotropia formed due to macular translocation. In 8 cases, superior and inferior oblique muscle surgery was combined with opposite vertical transpositions of the horizontal recti. In 5 cases, full tendon transposition of the superior oblique muscle to the nasal part of the globe was combined with the anterior margin advancement of the inferior oblique muscle. In 2 cases, only combined surgery of superior and inferior oblique muscles was performed. The degree of cyclodeviation was measured by both objective and subjective methods before and after counterrotation. Mean follow-up period was 53.9 days. RESULTS: The average objective cyclodeviation formed after macular translocation was 29.6 degrees. Combined surgery of superior and inferior oblique muscles led to a mean excyclorotation of 15 degrees whereas combined oblique muscle surgery performed with vertical transposition of horizontal recti provided a mean excylorotation of 20.8 degrees. The most effective method was the combination of total nasal transposition of superior oblique muscle with the anterior margin advancement of the inferior oblique muscle resulting in an improvement by 33 degrees. CONCLUSIONS: Macular translocations cause cyclodeviation. The ensuing cyclotropia can be eliminated by various surgical techniques of external eye muscles according to the degree of cyclotropia. PMID- 11932812 TI - [Bilateral necrobiotic xanthogranuloma of the eyelids with associated paraproteinemia: Case report and review of literature]. AB - BACKGROUND: Necrobiotic xanthogranuloma is a rare disorder which predominantly affects the periorbital region and is frequently associated with paraproteinemia and lymphoproliferative diseases. PATIENT AND METHODS: A 49-year old woman presented with bilateral, yellowish, subcutaneous lesions involving all four eyelids. These lesions have developed over the last year. Additionally, similar skin lesions developed on the trunk and the lower extremities. Two years ago, the patient had a bilateral episcleritis. An incisional biopsy was taken form the lesion. In addition, we reviewed the literature concerning similar cases. RESULTS: Histologic examination revealed a granulomatous process consisting of histiocytes, foamy cells, multinucleated giant cells (touton- and foreign body type) and associated with necrobiotic collagen and cholesterol clefts. General examination revealed the presence of a paraproteinemia of the IgG-typ associated with a hyperlipidemia. The patient was treated with a low dose chemotherapy which resulted in improvement of signs and symptoms. We could identify in the literature 75 similar cases. The mean age of those patients was 53 years. CONCLUSION: The clinical and histologic findings in our patient were typical for a necrobiotic xanthogranuloma with associated paraproteinemia. The most favorable treatment response has been reported with low doses of systemic administered chemotherapeutic agents in combination with radiation therapy; however, the mortality due to underlying diseases is high. PMID- 11932814 TI - [Publication languages of Impact Factor journals and of medical bibliographic databanks (reprint)]. AB - BACKGROUND AND OBJECTIVE: A preference for English-language sources during determination of Journal Impact Factors (IF) was discussed, IF being published in the annual Journal Citation Reports (JCR). The JCR are derived from data in Science Citation Index (SCI). The aim of this study was, therefore, (i) to review publication countries and languages in JCR, (ii) publication languages in SCI in comparison to further recognised medical bibliographic databanks. METHODS: Searching (i) countries and languages in JCR Science-Editions 1997 and 1998, (ii) language distributions in publication years 1995 - 2000 in bibliographic databanks SCI, MEDLINE (ME) and EMBASE (EM). RESULTS: (i) Almost 70 % journals in JCR 1997 and 1998 were published in USA, United Kingdom, or The Netherlands. Of two language options present, a number of English-classified journals contained > 90 % articles in other languages, whereas > 90 % publications in English could occur in Multi-Language (ML) journals, thereby complicating statistical comparisons. 83,9 % JCR-periodicals in 1997 and 85,6 % in 1998 were classified English. English/ML ratios increased exponentially with increasing IF. (ii) 95,5 % of the articles documented 1995 - 2000 in whole SCI and in our constructed SCI segment "Medicine and related areas" were written in English, compared to 88,5 % in ME and 89,8 % in EM. The SCI Medicine segment was 15 % more comprehensive than either MEDLINE or EMBASE. Highly significant differences of language distributions in SCI vs. MEDLINE and especially SCI vs. EMBASE were observed. Retrieval rates in SCI of German-, French-, Japanese- and Chinese-language medical papers published in 2000 were impressively augmented by EMBASE and MEDLINE. CONCLUSIONS: (i) Anglo-American publishers" countries and English language journals prevail in JCR with respect to numbers and IF levels. Publication language English favours citation frequency. (ii) Of databanks studied, SCI shows a maximum preference for English-language sources, thereby causing an English Language Bias during IF derivation. PMID- 11932813 TI - [Accidental injury of medial rectus muscle in endoscopic surgery of ethmoidal sinus: case report and survey of literature]. AB - BACKGROUND: Ophthalmologic complications of endoscopic sinus surgery are rare, consequences may be serious. PATIENTS: Two patients are presented who suffered lesions of the internal rectus muscles during endoscopic ethmoidectomy. The clinical picture of this rare complication resembles that of a paralysis of the medial rectus muscle. THERAPY AND COURSE: In one patient the very thinned medial rectus muscle was tucked in a first surgical step. In the second step the external rectus muscle was recessed. Fusion in primary gaze was possible. In the second patient surgery was planned, he died however. CONCLUSION: Because of the anatomy the medial rectus muscle is endangered in endoscopic ethmoidectomy. Considered to the frequency of this surgery the incidence however is rare. PMID- 11932815 TI - [Citation rates of medical German-language journals in English-language papers- do they correlate with the impact factor, and who cites? (reprint)]. AB - BACKGROUND AND OBJECTIVE: Several publications are warning that the German language is no longer needed for transmission of scientific data. One of the causes may be the Impact Factor (IF), which appears to be derived predominantly from Anglo-American journals. The aim of this study was to check actual international attention paid to German-language journals, i. e. their citation frequencies in English-language papers. Are these citing rates in English language articles correlated to the IF, and from where do citing articles originate? METHODS: Of 25 arbitrarily selected > 85 % German-language medical journals, IF as well as language distributions of citing articles were determined by searching publication years 1995 - 2000 in Science Citation Index (SCI). MEDLINE and EMBASE were used as supplementary retrieval systems. RESULTS: (i) The sample journals displayed an average IF = 0.357. A 99 % correlation (Pearson factor r = 0.987; n = 25) was observed between our "constructed" IF 2000 and IF published in Journal Citation Report 2000. This proves Stegmann's IF determination method to be valid. On the average, 53 % German-language and 45 % English-language articles between 1995 - 2000 cited the 1995 - 1999' contributions of the studied journals. No correlation was observed between IF vs. rates of citing articles in English (r < 0.1). 64 % of citing English-language articles showed corporate sources in Germany/Austria/Switzerland, and 13.5 % authors' institutions in USA. CONCLUSIONS: (i) An IF >/= 1 is, obviously, very hard to attain by German-language journals. ISI's differentiation between Citing vs. Cited-only Journals (the latter often serving as MEDLINE/EMBASE sources) during derivation of IF appears unjustified. (ii) English now serves as the predominant communication language in sciences in German-speaking countries, but has not supplanted the German language. Our study reveals remarkable international attention rates remaining. PMID- 11932816 TI - An in vitro study to evaluate the accuracy of stereotactic localization using magnetic resonance imaging by means of the Leksell stereotactic system. AB - The advantages of using magnetic resonance imaging (MRI) as opposed to computed tomographic (CT) scans or ventriculography in stereotactic surgery include the increased tissue contrast of the lesion or target, direct non-reformatted multiplanar imaging and target coordinate determination as well as reduced imaging artefacts produced by the stereotactic frame. One disadvantage of MR stereotaxis, however, is the potential for anatomic inaccuracy due to equipment induced inhomogeneities of the magnetic field. The authors present an experimental study on an in vitro model to examine the accuracy of target localization using the Leksell stereotactic frame and MR imaging. Ten formalin fixed brains taken from patients who had died of non-neurological diseases were sealed in a properly modelled plaster-cast shell simulating the skull bone. These models were fixed in the Leksell stereotactic frame and high-field MR images were performed (Siemens Magnetom SP 1.5 Tesla, T1-weighted spin echo sequences, TR/TE 600/15 ms, slice thickness 2 mm, FOV 300 mm). Following electrocoagulation of different targets on both lentiforme nuclei, the localization and extension of the lesions were controlled by MRI. A gross-/histopathological verification was performed. This model allows a good representation of the anatomic structures without any artefacts. The postoperative MRI control and the pathological examination of the lesions matched well with the preoperatively defined targets. The correlation of coordinates and measurements obtained with the pathological studies were within a +/- 2 mm range in all cases. PMID- 11932817 TI - Error analysis in cranial neuronavigation. AB - Neuronavigation systems are now an important component of many modern neurosurgical treatment strategies. Their support facilities intraoperative orientation and makes neurosurgical operations more precise and less traumatic. Computer-aided neurosurgery is definitively not a temporary fashionable phenomenon, the concept of neuronavigation is here to stay. This report summarizes a ten-years-long experience and presents an error analysis of 108 failures (12.4 %) in a total of 874 image-guided cranial neurosurgical procedures with an arm-linked (mechanical) system and two different infrared-light emitting (optical) systems. The application of neuronavigation incurs multiple reasons for pitfalls because of the complex man-machine interface. Principally, we have to differentiate two types of errors: "machine made errors" due to soft- or hardware failure and "man made errors" generally, due to inadequate handling of the navigation system. The error analysis demonstrated that the so-called human interface plays the main role causing a high error rate. PMID- 11932818 TI - Stereotactic biopsies guided by an optical navigation system: technique and clinical experience. AB - Frame-based stereotactic biopsies are time-consuming procedures making necessary head fixation in a ring, explicit coordinate calculation and setting of the parameters. Frameless systems make many of these intermediate steps unnecessary, impose less mechanical restrictions regarding access to the lesions, and with slight modifications can be used to perform stereotactic biopsies. A special adaptation designed to fix the holder and the biopsy instrument is described. The neuronavigation optical tracking system of Radionics was used. CT scans were performed with 6 skin markers. Calibration was performed after head fixation in the Mayfield clamp. Mean calibration error was 2.19 +/- 0.81 mm. The light emitting diode holder of the pointer was fixed into 2 Leila arms and moved under visual control based on CT images. The target point was selected, Leila arms fixed and a burr hole performed. The cannula was introduced to the target, being fitted with a depth stop in a length identical to that of the pointer on the screen. If necessary a second trajectory was easily selected. 49 patients underwent a frameless stereotactic biopsy. All targets except seven were superficial or in the white matter. In selected cases the biopsies were taken from deep-seated lesions. A histological diagnosis was obtained in 100 % of the cases. Four cases deteriorated postoperatively, two of these related to intratumoral bleeding. Navigation is a simple and effective method to perform biopsies of superficial and relatively large lesions. Frame-based procedures are restricted to brainstem tumors and lesions less than 1.5 cm in diameter. PMID- 11932819 TI - A simple method of reapplying the Leksell stereotactic frame without any repetition of the CT scan. AB - A simple method to reapply the Leksell stereotactic frame using marked carbon pins is described. This method allows the reapplication in a quick and safe manner without repetition of the CT scan. The reproducibility was very high. PMID- 11932820 TI - A non-invasive communication device for the paralyzed. AB - An EEG-based communication system has been developed to re-establish communication in severely paralyzed patients who operate the device by generating shifts of their slow cortical potentials. Training to gain control over slow cortical potentials was based on visual feedback and operant conditioning strategies. The vertical movement of a graphic signal on a computer screen informs the patients about the course of their slow cortical potential amplitude. Positive slow cortical potential shifts move the cursor up, negative shifts move it down. These shifts are then translated into binary responses. When a patient has achieved reliable control over his/her slow cortical potential shifts, these responses can be used to select or reject items presented at the bottom of the screen. As learning processes and applications differ considerably between patients, the present paper describes the data from one patient with amyotrophic lateral sclerosis. After about three months of training, this patient gained stable, near-perfect control over his slow cortical potentials. This skill enabled him to operate a specially designed program to communicate messages to his caregivers. PMID- 11932822 TI - Fully endoscopic vascular decompression of the trigeminal nerve. AB - Microvascular decompression of the trigeminal nerve is an accepted and effective means of treating patients with trigeminal neuralgia in whom compression of the nerve by a vascular structure is implicated in the pathogenesis of the disease. The current standard technique uses the binocular operating microscope for all intra-operative visualization. Posterior fossa endoscopy has demonstrated that the endoscope provides more comprehensive views of the anatomy of the cerebellopontine angle than does the operating microscope. To date, endoscopy has only been used to supplement microscopy in cranial nerve decompression surgery. In this report, we describe our completely endoscopic surgical technique as we present the case of a patient with trigeminal neuralgia who underwent successful vascular decompression by this approach. Using this technique the offending vessel was separated from the nerve with minimal brain retraction or dissection of surrounding structures. This report represents the first documented case where the endoscope was used as the exclusive imaging modality for decompression of the trigeminal nerve. From our experience we conclude that the endoscope's superior visualization more accurately identifies neurovascular conflicts, and provides a comprehensive evaluation of the completeness of the decompression. Additionally, this new method minimizes the risks of brain retraction and extensive dissection often required for microscopic exposure. From this study we conclude that completely endoscopic vascular decompression represents the next step forward in the safe and effective surgical treatment of trigeminal neuralgia. PMID- 11932821 TI - Virtual endoscopy for planning neuro-endoscopic intraventricular surgery. AB - To evaluate the usefulness of virtual endoscopy (VE) in planning neuroendoscopic intraventricular surgeries, the technique was applied in 20 of 22 consecutive procedures. Thirteen endoscopic third ventriculostomies (ETV) in 12 patients, 3 endoscopic colloid cyst removals, 1 third ventricular arachnoidal cyst fenestration, 1 endoscopic ventricul-cysto-cisternostomy (suprasellar arachnoidal cyst), 1 endoscopic tumor biopsy, one third ventricular gross total tumor removal and 2 septostomies at the foramen of Monro due the septal occlusion were performed. Contrast medium-enhanced MR images (3DMPRAGE, Siemens, Germany) were semi-automatically segmented with a surface-rendering technique ("Navigator" software, General Electric Medical, Buc, France) to produce the virtual endoluminal views. Surgery was performed with cerebral ventriculoscopes by Wolf (Richard Wolf, Knittlingen, Germany). VE was feasible in all patients and the virtual endoscopic images were comparable with the real intraventricular views obtained by standard rod lens systems. After contrast medium administration intra and paraventricular vessels such as the thalamocaudate vein, the septal veins, the basilar artery and its branches (distal BA complex) and the choroid plexus were identified on the virtual endoscopic images. In 8 patients, the additional anatomic information provided by VE profoundly influenced surgical planning. VE provides the neurosurgeon with additional morphological information supporting the planning process of neuroendoscopic intraventricular surgeries, contributing to the safety of the procedures. PMID- 11932823 TI - A rare symptomatic presentation of ecchordosis physaliphora and unique endoscope assisted surgical management. AB - We report on the presentation, diagnosis, and surgical management of a rare symptomatic case of ecchordosis physaliphora, including the use of endoscopy as a valuable imaging device in its operative management. A 49-year-old male presented with a intradural extra-axial mass located to the left of the basilar artery in the prepontine space. The tumor was exposed via a transmaxillary transclival approach and resected under binocular microscopic visualization. Prior to and following resection, endoscopes were introduced into the surgical field to conduct anatomic surveys of the region and to assess the completeness of tumor removal. Ecchordosis physaliphora is an uncommon benign lesion originating from embryonic notochordal remnants. It rarely causes clinical symptoms due to its slow growth patterns. Although similarities between EP of the spheno-occiput and chordomas of the clivus make distinction obscure, differentiation is important. Differences in these lesions impact upon patient prognosis as well as therapeutic strategies. The use of endoscopy in the resection of this mass marks an innovative approach to intraoperative imaging of the clival region; improved visualization of the prepontine area allows for more accurate defintion of the surgical anatomy of the tumor and for thorough assessment of the completeness of tumor removal. PMID- 11932824 TI - Endoscope-assisted interlaminar removal of an ependymoma of the cauda equina. AB - In oder to reduce surgical trauma to the lumbar spine during a multilevel laminectomy procedure we performed a multilevel, bilateral and interlaminar approach, with microsurgical and endoscopic techniques for removal of a large ependymoma of the lumbar spine in a 33-year-old female patient. Complete tumor removal has been achieved, while at the same time, the major elements that form the posterior spinal column could be preserved. The follow-up time is 4 years. PMID- 11932825 TI - Endoscopy in the 15th century. AB - The aim of this study was to investigate the contributions of Serefeddin Sabuncuoglu (1385-1468?)--who was born and practiced in Amasya in northern part of central Anatolia--to the field of endoscopy. He was a humble, curious and intelligent surgeon, and also a calligrapher and a miniature artist. This remarkable pioneer used the classification of the Andalusian surgeon, Abdul-Kasim Kalaf Ibni Abbas El Zahravi, known as Albucasis of Cordoba (936-1013); but improved it by adding many important and major new observations and chapters. He described "a thin and hollow tube of almost 35 cm, which was made of silver" and used it to inspect the interior of the bladder. PMID- 11932826 TI - Endoscopic endonasal removal of an intra-suprasellar Rathke's cleft cyst: case report and surgical considerations. AB - An endoscopic endonasal approach was performed to remove an intra-suprasellar Rathke's cleft cyst. Rathke's cleft cyst are benign lesions, rarely diagnosed because they are often asymptomatic. To the best of our knowledge, at least 475 cases of Rathke's cleft cysts have been reported. They seem to arise from remnants of Rathke's pouch, an invagination of the stomodeum. A 52-year-old woman, complaining of bilateral frontal headaches, was operated on by using an endoscopic endonasal approach, for an intra-suprasellar tumor. The pre-operative diagnosis was non-functioning pituitary adenoma. Intra-operatively a creamy coloured viscous tissue was found. After the removal of the cyst contents and of the capsule, the suprasellar structures were seen well. The chiasmatic cistern, the chiasm, the pituitary stalk and the pituitary gland were visualized with 0 and 30 degree endoscopes. The pathological findings showed a well-differentiated cuboidal epithelium. The diagnosis was Rathke's cleft cyst. No post-operative complications were observed. The endoscopic technique was particularly suitable in this case, both for the Rathke's cleft features and for an excellent outcome. The Rathke's cleft cyst was easily removed by suction and the cyst wall was entirely removed with curettes and pituitary punches. The hypophysis was distinguished from the cyst and was preserved. The surgical manoeuvres were all done under direct visual control. The absence of nasal packing and of breathing difficulties made comfortable the post-operative outcome. Thus, the endoscopic endonasal approach can be considered the favourite technique in case of either intra- and/or suprasellar Rathke's cleft cysts. PMID- 11932827 TI - Neuroendoscopic third ventriculostomy in a patient with occluded foramen of Monro: a case report. AB - We treated recurrent hydrocephalus in a previously shunted patient by neuroendoscopic third ventriculostomy. A tear was noted in the septum pellucidum. As the foramen of Monro was found to be occluded, we first fenestrated the floor of the lateral ventricle and then performed third ventriculostomy through the fenestration. A tough membrane believed to be Liliequist's membrane isolated the space immediately beneath floor of the third ventricle from the cistern below. Cerebrospinal fluid pulsation appeared after fenestration of this membrane. These obstructions presumably resulted from congenital and/or inflammatory causes. In cases such as this, successful neuroendoscopic third ventriculostomy presents a challenge to surgical judgment. PMID- 11932828 TI - Acute interhemispheric subdural hematoma: two case reports and analysis of the literature. AB - Interhemispheric subdural hematoma (ISH) had been considered extremely rare until identification with imaging studies. Its natural history is still quite unknown in terms of potential origin and course. As a consequence of various controversies, there is still no clearly established treatment, particularly in regard to medical or surgical management. The best decision requires tailoring treatment to the individual patient according to his or her clinical condition. Two patients with acute interhemispheric subdural hematomas are reported, the controversial management of this rare entity is analysed. In reviewing current published cases of interhemispheric subdural hematoma, it seems that the outcome of an individual patient is not related to the therapeutic approach, but to the level of consciousness and the neurological condition on admission. Surgical and medical treatment indications are taken into account for management. A patient with ISH developed an extension of the hematoma to the convexity at 2 weeks of his clinical course, with a decline of his neurological condition. The hematoma was then evacuated through a parietal craniotomy with an uneventful postoperative course. Another case of ISH presented as headache and TIA, with spontaneous clinical improvement at 12 hours and with no decline in the patient's neurological condition. Management was conservative. In both cases the neurological examination was normal after 6 months. As in previously reported cases, the clinical and neurological condition of the patient on admission is crucial for the course of an ISH. Treatment strategies are based on the individual neurological response of each case and the risk-benefit ratio to decide on a medical or surgical approach. PMID- 11932829 TI - Latex aneurysms: a technical note. AB - We describe a new technique for manufacturing latex tubings with thin-walled aneurysms. The physical characteristics of the latex model with its compliance simulates important details such as shape, size, neck and the thin wall of an aneurysm. Therefore important aspects of neuroendovascular therapy could be simulated and investigated in-vitro. PMID- 11932830 TI - A new instrument facilitates the needle trephination procedure: technical note. AB - INTRODUCTION: Percutaneous needle trephination is a well known neurosurgical procedure. The aim of this study was to develop a new instrument, which allows a stable fixation of an 18-G spinal needle in order to improve handling and precision of percutaneous needle trephinations. METHODS AND INSTRUMENTATION: The needle stabilizer was designed in a T-shape fashion morphologically similar to a corkscrew. The length of the uncovered needle tip is adjustable for individual requirements. RESULTS: Using the new needle stabilizing device a total number of 18 percutaneous needle trephinations were performed for the following indications: subdural hematoma, epidural hygroma, bifrontal air accumulation, superficial tumor cyst, superficial brain abscess. No complications have been observed. DISCUSSION: Our experience using this device shows that the modified technique fulfils criteria for clinical acceptance such as simplicity, low risk, reliability and cost effectiveness. PMID- 11932831 TI - Bio- and agroterror: the role of the veterinary academy. PMID- 11932832 TI - Who is responsible? PMID- 11932833 TI - Using technology to take a step back. PMID- 11932834 TI - Outcomes assessment. PMID- 11932835 TI - Outcomes assessment planning: an overview with applications in health sciences. AB - This article provides a brief overview of the process of outcomes assessment and examples of its application in professional health science education. The article is intended to provide a background for other articles in this issue describing ongoing activities in outcomes assessment in veterinary education and for programs considering developing a plan. The focus is on health professions other than veterinary medicine. The paper is not intended to be a comprehensive review of outcomes assessment; rather, it provides examples and models that may help clarify the outcomes assessment movement and identify trends in health care education. PMID- 11932836 TI - Using alumni research to assess a veterinary curriculum and alumni employment and reward patterns. AB - RATIONALE FOR THE STUDY: The purpose of the study was to obtain an outcomes assessment of the professional degree program of a veterinary college and to determine whether recently published national gender differences were true for a program that has always been predominantly female. METHODOLOGY: A survey was developed and mailed to all alumni of the veterinary degree program at North Carolina State University. Anonymous responses were collected by an independent organization for summation. Results were expressed numerically where possible, although the survey included numerous opportunities for textual responses. Responses were stratified by year of graduation, in five-year increments, and by gender of the respondents. RESULTS: Sixty-one percent of graduates remained within North Carolina. While most of the prerequisite courses were considered useful preparation for the curriculum, physics and calculus were exceptions. Over half the alumni thought time allocations to specific courses were appropriate. However, there were substantial differences between courses. The alumni were satisfied with most aspects of the training program, although there were differences between various components. There were substantial gender differences in length of first employment, salaries, species treated, practice ownership and type of ownership, and career satisfaction. Most alumni work in small animal private clinical practice. Ninety-four percent were employed within three months of graduating. Ninety-five percent did no work with the state's predominant agricultural species. Nineteen percent of alumni were either unsure, probably would not, or definitely would not become a veterinarian if they could choose again. CONCLUSION: The survey was a tremendously valuable source of information. Results provided support for curricular revision. The survey also provided comparative data in relation to national norms, where such norms were available. Unfortunately, few reports of this type are available, making inter-institutional comparisons difficult. The Association of American Veterinary Medical Colleges could assist with standardizing the process of outcomes assessment. PMID- 11932837 TI - Outcomes assessment in veterinary medical education. AB - The Virginia-Maryland Regional College of Veterinary Medicine (VMRCVM) agreed to perform outcomes assessment (OA) as part of the accreditation review process for the American Veterinary Medical Association (AVMA). Nine OA instruments were developed and validated by a 20-member accreditation committee. The instruments were also pre-tested by a subset of the target population. The instruments were for alumni one to five years post-graduate, alumni 6-15 years post-graduate, faculty, staff, DVM students, employers of veterinarians, referring veterinarians using the Blacksburg campus, and referring veterinarians using the Leesburg campus. In addition, data were used from OA surveys previously done for the Office of Research and Graduate Studies. Data from the surveys were incorporated into each of the 11 Essentials for Accreditation required by the AVMA. The process of OA provided a comprehensive assessment of the many aspects of the operation of the college. An important follow-up to the OA process is use of data to enhance and/or re-prioritize existing programs. PMID- 11932838 TI - Outcomes assessment at Tufts University School of Veterinary Medicine. PMID- 11932839 TI - Defining the attributes expected of graduating veterinary medical students, part 2: external evaluation and outcomes assessment. AB - We have previously defined a set of 62 attributes-12 in the area of professional characteristics, 28 addressing knowledge and understanding, and 22 delineating skills-that veterinary students should be expected to have demonstrated by the time of their graduation (Walsh DA, Osburn BI, Christopher MM. Defining the attributes expected of graduating veterinary medical students. J Am Vet Med Assoc 219:1358-1365, 2001). We have used this set of attributes as the basis of an outcomes assessment completed by California practitioners to determine whether graduates from the University of California School of Veterinary Medicine are meeting these expectations. Based upon this assessment, these 62 defined attributes appear to reflect very well practicing veterinarians' views and expectations of DVM graduates. The survey results also indicate that, overall, the recent University of California graduates are meeting these set of expectations. Simultaneously, the outcomes assessment focused attention on several areas, including private practice management, work expectations for successful practice, and surgical capabilities. For each, California practitioners recommended that the definition of the expectation be expanded and that the level of achievement by graduates be improved. Defining a set of attributes expected of veterinary graduates is a key step in obtaining an effective outcomes assessment of a professional educational program. PMID- 11932840 TI - Group learning improves case analysis in veterinary medicine. AB - INTRODUCTION: Group learning has become important to professional students in the healing sciences. Groups share factual and procedural resources to enhance their performances. METHODOLOGY: We investigated the extent to which students analyzing case-based evaluations as teams acquired an immediate performance advantage relative to those analyzing them as individuals and the extent to which group work on one problem led to better performance by individual students on related problems. We blinded written evaluations by randomly assigning numbers to groups of students and using removable tracers. Differences between groups and individuals were evaluated using Student's t statistic. Similar comparisons were evaluated by meta-analysis to determine overall trends. RESULTS: Students who analyzed evaluations as a group had an 8.5% performance advantage over those who analyzed them as individuals. When evaluations were divided into those asking questions related to treatment, differential diagnosis, and prognosis, specific performance advantages for groups relative to individuals were 8.9%, 5.9%, and 6.1% respectively. Students who had previously been trained by group evaluations had a 1.5% advantage relative to those who received their training as individuals. CONCLUSIONS: Answers by students analyzing evaluations as groups suggested a deeper understanding, in large part because of their improved ability to explain treatment and to conduct differential diagnosis. These improvements suggested limited abilities to use previous experience to improve present performance. PMID- 11932841 TI - Surgical education: attitudes toward animal use in teaching surgery at Louisiana State University. AB - Concerns over the use of animals in teaching have lead to a reduction in the number of live animal laboratories. A survey of 275 students and faculty was conducted to characterize their attitude toward the use of animals in teaching surgery. Respondents favored live animal use. They believed that model laboratories were helpful in learning aseptic technique and suturing skills but less helpful in learning tissue handling, dissection, hemostasis, or anesthesia. PMID- 11932842 TI - An interactive, student-centered approach, adopting the SOLO taxonomy, for learning to analyze laboratory data in veterinary clinical pathology. AB - INTRODUCTION: The purpose of this study was to describe and evaluate an interactive, student-centered teaching strategy for learning to analyze laboratory data in veterinary clinical pathology. The strategy was designed to operate in tutorials of approximately one hour duration and adopted the structure of the observed learning outcome (SOLO) taxonomy in order to align with outcomes and assessment components of unit of study design and to encourage a deep approach to learning. METHODOLOGY: The teaching strategy adopted group discussion and reflective observation as core activities. Students worked alone in identifying abnormal laboratory data, in pairs in discussing possible reasons for the abnormalities, and in two larger groups in deciding on conclusions for, and further investigation and management of, the case. The final debriefing brought the two groups together to reflect, question, and reach a consensus about the case. The teaching strategy was evaluated on the basis of its success in encouraging interaction through discussion, developing self confidence in analyzing laboratory data, and enhancing understanding as to how the disciplines of veterinary clinical pathology and veterinary medicine interrelate. Evaluation used self-reflection, peer feedback, and a student questionnaire. CONCLUSION: The teaching strategy provided the opportunity for students to develop and practice an approach to the analysis of laboratory data in a manner consistent with current educational thinking on student-centered learning. The use of group discussion and significant reflective practice not only enhanced interpersonal skills but also encouraged a deep approach to learning, leading to ownership of knowledge and increased awareness of the worth of veterinary clinical pathology in the investigative process. PMID- 11932843 TI - Upper-extremity torque production in men with paraplegia who use wheelchairs. AB - OBJECTIVES: To study maximal upper-extremity torque values and agonist/antagonist torque ratios in a sample of individuals with paraplegia and to compare these values with a sample of age-, gender-, and weight-matched able-bodied individuals. DESIGN: Cross-sectional, case-control study. SETTING: A biomechanics laboratory. PARTICIPANTS: A convenience sample of 15 men with complete (American Spinal Injury Association class A) T5-L2 paraplegia and 15 able-bodied control subjects. INTERVENTIONS: Not applicable. MAIN OUTCOME MEASURES: Maximal isometric torques of 12 upper-extremity muscle groups (shoulder, elbow, and wrist flexion and extension; shoulder abduction and adduction; shoulder internal and external rotation; elbow pronation and supination) were measured with custom dynamometers. RESULTS: Maximal torque values and agonist/antagonist torque ratios for the shoulder, elbow, and wrist functions did not differ between the control subjects and those with complete paraplegia, with the exception of the supination/pronation torque ratio, which was significantly higher in the able bodied control group. CONCLUSION: Wheelchair propulsion and other functional activities, such as transfers, do not significantly affect maximum torque production of the upper extremities in individuals with paraplegia. On the basis of these findings, arguments that upper-extremity muscle imbalances are important contributory factors to upper-extremity pain and dysfunction in this group may be too simplistic. PMID- 11932844 TI - A new 3-point bending traction method for restoring cervical lordosis and cervical manipulation: a nonrandomized clinical controlled trial. AB - OBJECTIVE: To evaluate a new 3-point bending type of cervical traction. DESIGN: Nonrandomized controlled trial of prospective, consecutive patients compared with control subjects. Follow-up patient data were obtained at 3 and 15(1/2) months, and 8 1/10 months for controls. SETTING: Data were collected at a spine clinic in Nevada. PATIENTS: Volunteer subjects consisted of 30 patients and 24 controls. Subjects had cervicogenic pain (neck pain, headaches, arm pain, and/or numbness). Subjects were included if their Ruth Jackson radiographic stress lines measured less than 25 degrees but were excluded if they had suspected disk herniation or canal stenosis. All subjects completed the first follow-up examinations, and 25 of 30 patients completed the long-term follow-up examination. INTERVENTIONS: Spinal manipulation for pain and a new form of 3-point bending cervical traction to improve lordosis. Cervical manipulation was provided for the first 3 to 4 weeks of treatment. Traction treatment consisted of 3 to 5 sessions per week for 9 +/- 1 weeks. MAIN OUTCOMES MEASURES: Besides pain visual analog scale (VAS) ratings, pre- and posttreatment lateral cervical radiographs were analyzed. RESULTS: Control subjects reported no change in the pain VAS ratings and had no statistically significant change in segmental or global radiographic alignment. For the traction group, VAS ratings were 4.3 pretreatment and 1.6 posttreatment. Traction group radiographic measurements showed statistically significant improvements (P <.008 in all instances of statistical significance), including anterior head weight bearing (improved 6.2mm), Cobb angle at C2-7 (improved 12.1 degrees ), and angle between posterior tangents at C2-7 (improved 14.2 degrees ). For the treatment group, at 15(1/2)-month follow-up, only minimal loss of C2-7 lordosis (3.5 degrees ) was observed. CONCLUSIONS: Sagittal cervical traction with transverse load at midneck (2-way cervical traction) combined with cervical manipulation can improve cervical lordosis in 8 to 10 weeks as indicated by increases in segmental and global cervical alignment. Magnitude of lordosis at C2 7 remained stable at long-term follow-up. PMID- 11932845 TI - Objective measurement of muscle strength in children with spastic diplegia after selective dorsal rhizotomy. AB - OBJECTIVES: To examine changes in isometric muscle strength at the elbow, knee, and ankle at 6 months and 1 year after selective dorsal rhizotomy (SDR) and to determine if SDR altered the frequency of muscle cocontraction. DESIGN: Prospective outcome study of a consecutive sample. SETTING: Children's hospital. PATIENTS: Ten children with spastic diplegia (7 independent and 3 dependent ambulators who used assistive devices) and 8 age-matched controls. INTERVENTIONS: SDR; physical and occupational therapy; elbow, knee, and ankle measured for flexion and extension strength during three 10-second isometric contractions for each muscle group; and monitored cocontraction measured via muscle electrodes. MAIN OUTCOME MEASURES: Absolute and normalized values of isometric strength; and alterations in the frequency of cocontraction at 6 months and 1 year postoperatively. RESULTS: Children with spastic diplegia showed significantly weaker knee extensors, ankle dorsiflexors, and ankle plantarflexors than age matched controls. There were no significant differences in strength between the 2 groups in the elbow flexors, elbow extensors, and knee flexors. Isometric strength did not increase or decrease significantly after SDR. Cocontraction during knee extension was normalized after SDR, whereas cocontraction during ankle plantarflexion was unchanged by SDR in the majority of children. CONCLUSION: SDR did not result in a significant decrease in muscle strength in ambulatory children with spastic diplegia. The normalization of the electromyographic patterns at the knee and not the ankle after SDR lends support to the premise that in children with cerebral palsy cocontraction is multifaceted, representing a volitional strategy to enhance control, as well as a disorder of the mechanisms that govern patterns of muscle activity. PMID- 11932846 TI - A descriptive study of psychiatric disorders and psychosocial burden in rehabilitation patients with musculoskeletal diseases. AB - OBJECTIVE: To investigate current, 12-month, and lifetime prevalence rates, and associated psychosocial burden of psychiatric disorders in rehabilitation inpatients with musculoskeletal diseases. DESIGN: Two-stage epidemiologic survey. SETTING: Four orthopedic rehabilitation inpatient clinics in southwest Germany. PARTICIPANTS: A total of 910 inpatients with different musculoskeletal diseases participated in the survey. According to their General Health Questionnaire-12 scores, 205 patients were selected randomly for standardized interviews. INTERVENTIONS: Not applicable. MAIN OUTCOME MEASURES: Psychosocial burden (Hospital Anxiety and Depression Scale, Lubeck Alcoholism Screening Test) and quality of life (Medical Outcomes Study 36-Item Short-Form Health Survey), assessment of diagnosis and somatic parameters through standardized medical records. Clinical interview (Munich Composite International Diagnostic Interview) in the second-stage examination to obtain Diagnostic and Statistical Manual of Mental Disorders (4th edition) diagnoses of psychiatric disorders. RESULTS: Prevalence rates of psychiatric disorders are 31.1% for the 4-week period, 47.1% for the 12-month period, and 64.6% for the lifetime period. The most prevalent current disorders are anxiety (15%), affective (10.7%), and substance-related disorders (9.2%). Half of the comorbid ill patients have 2 or more simultaneous psychiatric disorders and report elevated levels of psychosocial burden (eg, intense pain, low quality of life, more days of sick leave). CONCLUSIONS: Patients undergoing musculoskeletal rehabilitation should be assessed carefully for comorbid psychiatric illnesses. Further research should be undertaken to evaluate the effectiveness of psychosocial interventions for comorbid psychiatric disorders on life quality, therapeutic compliance, and outcome of rehabilitation treatment. PMID- 11932847 TI - Vision, attention, and self-reported driving behaviors in community-dwelling stroke survivors. AB - OBJECTIVE: To elucidate the relationships among vision, attention, driving status, and self-reported driving behaviors in community-dwelling stroke survivors. DESIGN: A cross-sectional design to compare stroke survivors to older adults without stroke on visual measures, attentional measures, and self-reported driving behaviors. SETTING: Rehabilitation center at a university hospital. PARTICIPANTS: Fifty stroke survivors and 105 older adults without neurologic or visual impairment. INTERVENTIONS: Not applicable. MAIN OUTCOME MEASURES: Visual acuity, contrast sensitivity, peripheral vision, useful field of view (UFOV), Behavioral Inattention Test, and a driving habits questionnaire. RESULTS: Stroke survivors had impaired contrast sensitivity, peripheral vision, and UFOV compared with older adults in good visual and neurologic health. Driving stroke survivors typically had less attentional impairment than nondrivers. Stroke survivors who returned to driving reported difficulty in challenging driving conditions, drove less, and relied more on other people for transportation than older adults without stroke. CONCLUSIONS: These results suggest that vision and attention, both of which are important for driving, are often impaired in stroke survivors. The severity of these deficits could be an influence on driving status and driving behavior. Stroke survivors who return to driving strategically limit their driving exposure and rely on others for transportation, which suggests that they may deliberately self-regulate their driving behavior. PMID- 11932848 TI - The effect of shoe wedges and lifts on symmetry of stance and weight bearing in hemiparetic individuals. AB - OBJECTIVE: To determine the effect of shoe wedges and lifts on symmetry of stance and weight bearing in hemiparetic individuals. DESIGN: Weight bearing on the paretic side was measured in patients with hemiparesis during quiet standing and in conditions of compelled weight shift. SETTING: Free-standing acute inpatient rehabilitation hospital. PARTICIPANTS: Nine individuals with hemiparesis as a result of unilateral stroke who were able to stand for 3 to 5 minutes without assistance or rest, and satisfied other inclusion criteria. INTERVENTIONS: Compelled shift of the body weight was induced with different shoe wedges (5 degrees, 7.5 degrees, 12.5 degrees) or shoe lifts (0.6, 0.9, 1.2cm), which extended under the entire shoe of the unaffected limb. Weight-bearing symmetry scores were used to characterize the symmetry of stance. MAIN OUTCOME MEASURES: Weight-bearing symmetry scores. RESULTS: Without a shoe wedge or a shoe lift, weight-bearing symmetry was characterized by underloading of the paretic limb (39.90% +/-.80% of body weight). Weight shift induced by shoe wedges or shoe lifts applied to the unaffected limb promoted improved symmetry of weight bearing and stance. A shoe wedge of 5 degrees provided the most symmetrical weight distribution (51.44% +/- 1.88% of body weight). CONCLUSION: Shoe wedges and shoe lifts under the unaffected limb induced compelled weight shift toward the paretic limb, resulting in improved symmetry of stance of individuals with mild hemiparesis. We suggest that improved symmetry of bipedal standing obtained with a shoe wedge or a shoe lift applied to the unaffected limb can help overcome the learned disuse of the affected limb. We further suggest that weight distribution induced by shoe wedges or shoe lifts may help in the treatment of ambulatory hemiparetic individuals with asymmetrical stance caused by unilateral stroke. PMID- 11932850 TI - Resistive inspiratory muscle training in sleep-disordered breathing of traumatic tetraplegia. AB - OBJECTIVE: To assess the effect of resistive inspiratory muscle training (RIMT) on the static pulmonary function and sleep-induced breathing disorder of individuals with chronic cervical cord injury. DESIGN: Before-after training. SETTING: Home-setting training program. PATIENTS: Fourteen complete traumatic tetraplegic patients (12 men, 2 women; mean age, 41.1 +/- 14y; range, 19-56y) injured for more than 6 months. INTERVENTION: Subjects participated in a 6-week RIMT program for 15 minutes twice daily at a training intensity of 60% of maximum inspiratory pressure (MIP). The participants were reevaluated at the end of 6 week training. MAIN OUTCOME MEASURES: Lung volume, peak expiratory flow (PEF), MIP, and maximum expiratory pressure (MEP) were measured by using a spirometry and inspiratory force meter, respectively. Capnography was used to monitor nocturnal pulse oxyhemoglobin saturation (SpO(2)) and end-tidal carbon dioxide tension level (ETCO(2)) of the patients. RESULTS: The maximum voluntary ventilation (MVV) and MIP of individuals with chronic cervical cord injury substantially improved after RIMT. MIP increased from -68.7 +/- 27.4cmH(2)O to 77.3 +/- 24.0cmH(2)O and MVV rose from 62.7 +/- 33.2L to 73.4 +/- 31.3L (P <.05). Despite increasing from 3.5 +/- 1.8L/s to 4.0 +/- 1.7L/s, PEF was statistically insignificant. For the individuals with improved MIP, the duration of ETCO(2) greater than 48mmHg reduced from 2.2% +/- 3.3% to 1.0% +/- 2.0% of total sleep time (P =.05) and that of SpO(2) less than 90% significantly declined from 1.8% +/- 2.8% to 1.3% +/- 2.4% of total sleep time (P <.05). CONCLUSION: These findings suggest that RIMT can enhance the respiratory muscle strength and endurance of chronic tetraplegia and further ameliorate the sleep-induced breathing disorder. Therefore, RIMT is suggested as a home program for patients with sleep-disordered breathing. PMID- 11932849 TI - Mobility status during inpatient rehabilitation: a comparison of patients with stroke and traumatic brain injury. AB - OBJECTIVES: To compare the mobility status (admission and discharge status, change in status) between patients with stroke and traumatic brain injury (TBI) during inpatient rehabilitation and to determine the relationship between mobility status and outcome variables including length of stay (LOS). DESIGN: Prospective study. SETTING: Free-standing tertiary rehabilitation center. PARTICIPANTS: A total of 210 patients with stroke (n = 136) and TBI (n = 74) consecutively admitted for inpatient rehabilitation. INTERVENTIONS: Not applicable. MAIN OUTCOME MEASURES: Clinical Outcome Variable Scale (COVS), a 13 item scale of mobility status (measured on admission and discharge from inpatient rehabilitation), and rehabilitation LOS. RESULTS: With age and time since injury controlled in the model, the TBI group showed a significantly higher mobility status on admission and discharge over the stroke group, but the change (improvement) in mobility status did not differ. The admission mobility status accounted for 61% and 60% of variability of the discharge mobility status for the stroke and TBI groups, respectively. The admission mobility status accounted for 40% and 50% of the variability in rehabilitation LOS for the stroke and TBI groups, respectively. Either the admission mobility status or the physical therapist's prediction of the discharge status could be used to determine the actual discharge mobility status, although the physical therapist's predictions were more accurate than using a statistical model. CONCLUSIONS: The TBI group showed a higher mobility status at admission and discharge from inpatient rehabilitation than the stroke group; however, the rate of improvement (improvement in mobility status per day) did not differ between groups. Admission mobility status using the COVS was an excellent predictor of discharge mobility status and rehabilitation LOS in stroke and TBI patients. PMID- 11932851 TI - Reliability and validity of combined imaging and pressures assessment methods for diabetic feet. AB - OBJECTIVES: To determine the reliability of using 3-dimensional (3D) spiral x-ray computed tomography (SXCT) imaging methods to measure anatomic foot structure and to test the validity of using a loading device to simulate walking pressures on the plantar foot during SXCT examination. DESIGN: Nonrandomized control trial of consecutive patients with complete data compared with age-matched controls. SETTING: A clinical SXCT scanner and physical therapy laboratory in a local hospital. PARTICIPANTS: Eight subjects with diabetes and a history of forefoot ulcers and 8 control subjects. INTERVENTIONS: SXCT imaging and plantar pressure analysis were used to acquire 3D volumetric structure and pressure data of the foot during 4 foot positions, 2 weight bearing and 2 non-weight bearing. MAIN OUTCOME MEASURES: Differences between repeated SXCT measures of foot structure. Comparisons of plantar foot pressure. Foot structure measurements. Methods for measuring the bony angles and soft-tissue thickness were developed and reliability tests were performed. RESULTS: There was essentially no bias (<+/ 0.5mm or +/-0.5 degrees ) and a high degree of reliability (81% of the measures had a reliability <2.0mm or 2 degrees ) when measuring foot structures. Correlations of pressure measures collected on the loading device compared with those collected during walking were (mean +/- standard deviation) r =.66 +/-.06. The percentage agreement between the pressures collected on the loading device and during walking was 91.1% +/- 4.7%. CONCLUSION: Reliability of anatomic foot structure measurements and validity of plantar loading during visualization were good. These methods may be used to determine structural differences between diabetic and healthy feet and to evaluate how these differences relate to plantar pressures. PMID- 11932852 TI - Analysis of FIM instrument scores for patients admitted to an inpatient cardiac rehabilitation program. AB - OBJECTIVE: To evaluate in an inpatient cardiac rehabilitation program (a phase IB) whether length of stay (LOS), discharge to home, and improvement in physical function differed between patients with lower and higher degrees of functional independence on admission. DESIGN: A retrospective study. SETTING: A public acute long-term care hospital. PATIENTS: All cardiac rehabilitation patients (N = 143) admitted to the hospital from January 1998 through June 1999. Patients were divided into a higher- and a lower-functioning group by using the admission FIM instrument scores above and below the midpoint of 72. Comparisons in LOS, discharge disposition, and functional gains between these 2 groups were then performed. INTERVENTIONS: Not applicable. MAIN OUTCOME MEASURES: FIM scores, FIM change, FIM gains per week, LOS, and discharge disposition. RESULTS: Total FIM scores at discharge were significantly higher than those on admission (25%, P <.0001). The median value of total FIM gains per week was 7.78 with a stay of 17 days and a home discharge rate of 76%. The higher-functioning group (n = 106) differed from the lower group (n = 37) with shorter stay (15 vs 23d, P <.0001), greater FIM gains per week (8.6 vs 4.8, P =.002), and greater likelihood of discharge to home or community (84% vs 54%, P <.001). The average incremental FIM change in each group was the same. In multivariate analysis, both admission (P =.001) and discharge (P <.001) FIM scores were the best predictors of patients' discharge disposition to home. CONCLUSIONS: Admission FIM scores are important predictors for the clinical course and discharge outcomes of cardiac rehabilitation patients, with those with higher admission FIM scores having a shorter LOS and greater likelihood of discharge to home. The admission FIM scores can help to establish realistic goals. PMID- 11932853 TI - Footdrop, foot rotation, and plantarflexor failure in Charcot-Marie-Tooth disease. AB - OBJECTIVE: To evaluate the frequency of occurrence of the main causes of poor stance and gait in patients with Charcot-Marie-Tooth disease (footdrop graded as mild or severe, rotation, and plantarflexor failure) both as single and associated problems. DESIGN: Observational. SETTING: A neuromuscular disorders department in a specialized Italian rehabilitation hospital. PARTICIPANTS: One hundred twenty-six nonoperated lower limbs from 64 outpatients. INTERVENTIONS: Ankle angle during active dorsiflexion and heel angle in stance were measured in a photograph; the ability to raise the heel at least 2cm was evaluated. MAIN OUTCOME MEASURES: Frequency of mild footdrop (ankle angle < or =100 degrees ), severe footdrop (ankle angle >100 degrees ), and rotation and plantarflexor failure singularly and in association. RESULTS: Mild footdrop was present in 47.6% of examined limbs and was associated with rotation in 39.7% of limbs; severe footdrop was present in 52.4% of limbs and was associated with rotation in 28.6% of limbs, with rotation plus plantarflexor failure in 21.4% of limbs. CONCLUSIONS: We recommend categorizing problems in the lower limbs into 4 levels of increasing symptom severity, starting with mild footdrop and graduating to the instance where the 3 problems are associated. PMID- 11932854 TI - Substance abuse attitudes and policies in US rehabilitation training programs: a comparison of 1985 and 2000. AB - OBJECTIVE: To assess and compare the attitudes, beliefs, and policies of physical medicine and rehabilitation (PM&R) training programs toward substance abuse and tobacco use over the last 15 years. DESIGN: A blinded questionnaire was sent to all US rehabilitation medicine training program directors. Results were compared with a survey conducted in 1985. SETTING: US PM&R residency training programs with inpatient rehabilitation training. PARTICIPANTS: Training directors or their designated agents. INTERVENTION: A 35-item questionnaire was mailed between November 1999 and April 2000 to the 81 US training programs identified by the American Board of Physical Medicine and Rehabilitation as having rehabilitation training programs with inpatient rehabilitation units. Responses were pooled by our Survey Research Center to preserve anonymity. Training programs that did not respond received additional mailings and telephone calls to improve the response rate. MAIN OUTCOME MEASURES: Chi-square analysis to assess changes in responses with time. RESULTS: Forty-six of the 79 (58%) eligible training programs responded (1 program had merged, 1 did not provide inpatient rehabilitation). Programs were located in cities ranging from less than 100,000 (n = 2) to greater than a million inhabitants (n = 18). Eighty percent (37/46) of the respondents were "concerned or very concerned" about their patients' alcohol and drug use, and 69% routinely assessed patients for alcohol and drug use compared with only 25% in 1985 (P <.00001). Almost all respondents (43/46) supported written guidelines to prohibit alcohol and drug use by patients in the rehabilitation unit. Eighty-three percent had a prohibition policy, and 72% had written guidelines. Both of those rates represent increases from the 1985 response rates of 65% and 45%, respectively. Ambivalence persisted about appropriate treatment programs for persons with disabilities: in 1985, 51% of the respondents agreed that a person with a disability could be treated appropriately in a substance abuse program designed for persons without a disability; in 2000, the percentage had increased to 64%. All respondents believed that tobacco use is an addiction, but only 25% of their units offered tobacco cessation services to patients on their rehabilitation unit. CONCLUSION: The survey results are encouraging. Since 1985, not only have substance abuse issues been recognized, but also systemic institutionalized approaches (eg, regular screening, written guidelines) have increased markedly. Tobacco is now uniformly accepted as an addiction, but screening and access to cessation programs are similar to that available for alcohol and drug treatment programs 15 years ago. PMID- 11932855 TI - Comparison of knee extensor strength between limbs in individuals with bilateral total knee replacement. AB - OBJECTIVES: To determine and compare force production of the knee extensors between limbs after bilateral total knee arthroplasty (TKA). DESIGN: Two repeated measures designs (2 factors: limb, time) assessing for differences in strength between limbs over 3 times at 2 different testing speeds. SETTING: Private and institutional practice. PARTICIPANTS: Fourteen volunteers (6 men, 8 women; mean age +/- standard deviation, 74.57 +/- 5.5 y; height, 169.91 +/- 9.43 cm; weight, 76.42 +/- 9.85 kg) with bilateral TKA. Open-chain knee extensor strength was measured in both legs before surgery and 30 and 60 days after TKA at 60 degrees /s and 180 degrees /s. INTERVENTION: Rehabilitation 3 times weekly for 8 weeks. Treatment consisted of gait, transfer training, stationary cycle work, and a progressive resistance-training program. MAIN OUTCOME MEASURE: Peak torque measured isokinetically. RESULTS: Limb by time interaction was not significant for either 60 degrees /s (F(2,26) =.75, P =.351) or 180 degrees /s (F(2,26) =.78, P =.393). A significant main effect of time was found at 60 degrees /s (F(2,26) = 14.22, P =.0001) and 180 degrees /s (F(2,26) = 10.47, P =.003). CONCLUSIONS: During the first month after TKA, bilateral knee extensor force-production decreased significantly compared with preoperative force production. PMID- 11932856 TI - An ergonomic evaluation comparing desktop, notebook, and subnotebook computers. AB - OBJECTIVE: To evaluate and compare the postures and movements of the cervical and upper thoracic spine, the typing performance, and workstation ergonomic factors when using a desktop, notebook, and subnotebook computers. DESIGN: Repeated measures design. SETTING: A motion analysis laboratory with an electromagnetic tracking device. PARTICIPANTS: A convenience sample of 21 university students between ages 20 and 24 years with no history of neck or shoulder discomfort. INTERVENTION: Each subject performed a standardized typing task by using each of the 3 computers. MAIN OUTCOME MEASURES: Measurements during the typing task were taken at set intervals. RESULTS: Cervical and thoracic spines adopted a more flexed posture in using the smaller-sized computers. There were significantly greater neck movements in using desktop computers when compared with the notebook and subnotebook computers. The viewing distances adopted by the subjects decreased as the computer size decreased. Typing performance and subjective rating of difficulty in using the keyboards were also significantly different among the 3 types of computers. CONCLUSIONS: Computer users need to consider the posture of the spine and potential risk of developing musculoskeletal discomfort in choosing computers. PMID- 11932857 TI - Long-term follow-up of patients with nonparalytic poliomyelitis. AB - OBJECTIVE: To examine patients with previous nonparalytic poliomyelitis in search of muscle atrophy, weakness, and other late symptoms of poliomyelitis. DESIGN: A mailed questionnaire followed up with neurologic and neurophysiologic examinations of respondents who reported symptoms possibly related to the late sequelae of polio. SETTING: Neurology department at a university hospital. PARTICIPANTS: Thirty-nine of 47 patients diagnosed with nonparalytic poliomyelitis and hospitalized at a Norwegian hospital between 1950 and 1954, during the Norwegian polio epidemic. INTERVENTIONS: Not applicable. MAIN OUTCOME MEASURES: Electromyography to determine function of the anterior tibialis, vastus lateralis, and biceps brachii muscles; nerve conduction studies of the sural, peroneal, and tibial nerves; motor and sensory nerve conduction velocity, and compound muscle and sensory nerve action potentials, and distal latencies. RESULTS: Twenty-five of 47 patients (53.2%) reported symptoms possibly related to the late sequelae of poliomyelitis. Eight of 20 examined symptomatic patients had normal neurologic and neurophysiologic findings, whereas 9 others had other medical conditions that could explain the symptoms. Three patients (6.7%) had neurologic and neurophysiologic findings and development of symptoms consistent with motoneuron damage. CONCLUSION: Some nonparalytic patients may have subclinical acute motoneuron damage with subsequent development and manifestation of motor weakness and neuromuscular symptoms many years later. These symptoms should be considered a differential diagnosis in patients who have a history of nonparalytic poliomyelitis. PMID- 11932858 TI - Is the functional reach test useful for identifying falls risk among individuals with Parkinson's disease? AB - OBJECTIVE: To evaluate the effectiveness and validity of the Functional Reach Test (FRT) as a screening tool to identify fallers (persons at risk for falls) among subjects with Parkinson's disease (PD) and control subjects. DESIGN: A case comparison design with a consecutive sample. Subjects performed 3 consecutive functional reach trials. SETTING: Motor behavior laboratory in a university setting. PARTICIPANTS: Fifty-eight adults (43 subjects with PD, 15 control subjects). Controls were recruited from a Florida hospital and the local community. INTERVENTIONS: Not applicable. MAIN OUTCOME MEASURES: A falls history was recorded, a mean FRT score attained, and FRT scores were categorized as less than 25.4 cm, the criterion for falls risk, or > or =25.4 cm. RESULTS: Mean FRT scores differentiated subjects with PD and a known history of falls from subjects with PD and no history of falls and from control subjects (P <.001). Tests of validity for the FRT as a screening tool indicated sensitivity as 30%, specificity as 92%, positive predictive value as 90%, and negative predictive value as 36%. CONCLUSIONS: The FRT, using a reach less than 25.4 cm as a criterion for falls risk, is not a sensitive instrument for identifying individuals with PD at risk. However, the percentage of those persons identified as at risk by the FRT are highly likely at risk, and they should be referred for falls risk intervention. Because the FRT does not identify every person at risk, using a test battery addressing other factors contributing to falls risk may increase the sensitivity of a clinician's assessment to identify persons with PD at risk for falls. PMID- 11932859 TI - Skin temperature response to cryotherapy. AB - OBJECTIVES: To compare the localized skin-cooling effects of 2 cryotherapy modalities and to review the clinical relevance of the results. DESIGN: Randomized controlled trial with repeated measures. SETTING: Laboratory experiment. PARTICIPANTS: Convenience sample of 20 volunteers (13 women, 7 men), ages 21.3 to 44 years (mean, 31.3 +/- 6.8 y). INTERVENTIONS: A flexible frozen gel pack, a 454 g packet of frozen peas, or a control applied to the anterior thigh. No blinding was undertaken. MAIN OUTCOME MEASURE: Surface skin temperature under the modality at baseline and 10 and 20 minutes after application. RESULTS: Significant effects were recorded for modality (F(2) = 290.56, P <.0001), time (F(1.27) = 1868.07, P <.0001), and their interaction (F(2.09) = 305.47, P <.0001). After 20 minutes, frozen peas produced the lowest mean skin temperature +/- standard deviation of 10.8 degrees C +/- 2.28 degrees C compared with 14.4 degrees C +/- 2.53 degrees C from the gel pack and 26.1 degrees C +/- 1.75 degrees C from the control. Skin temperature fell between both time periods with the application of frozen peas but stabilized after 10 minutes of gel pack and control application. CONCLUSIONS: Application of frozen peas produced mean skin temperatures adequate to induce localized skin analgesia, to reduce nerve conduction velocity, and to reduce metabolic enzyme activity to clinically relevant levels. Flexible frozen gel packs did not cool skin sufficiently to achieve these levels. PMID- 11932860 TI - Preventing fatigue of fast striated muscles of the pelvic floor and slow striated muscles of the limb by manipulating the on-off time of electric stimulation. AB - OBJECTIVE: To investigate in vitro the effect of different on-off electrostimulation times on the fatigue of 2 pelvic muscles, the iliococcygeus and pubococcygeus (mainly fast), and the soleus muscle of the limb (mainly slow). DESIGN: Nonrandomized controlled trial and before-after trial. SETTINGS: In vitro study in tissue baths on stabilized preparations of whole striated muscles. ANIMALS: Young healthy female Wistar rats. INTERVENTION: Applied isometric tetanic contractions of various stimulation durations (on-off times) to dissected rat iliococcygeus and pubococcygeus (at 66Hz) and soleus (at 25Hz) muscles. MAIN OUTCOME MEASURE: Muscle fatigue with changing on-off, frequency, and stimulation times. RESULTS: The longer the off time, the less the fatigue. Shortening the stimulation time can minimize fatigue. Optimal combinations are presented to limit fatigue. CONCLUSION: Changing on-off time and stimulation time can be used to make electric stimulation of pelvic muscles more effective. PMID- 11932862 TI - Popliteal aneurysm as a source of phantom pain: a case report. AB - A 42-year-old man with a remote history of right transtibial amputation but no history of phantom pain developed severe phantom pain 10 years after amputation. A literature review suggested that his presentation was contrary to the natural history of phantom pain, which is usually most prominent in the early stages after amputation. Diagnostic workup revealed a popliteal artery aneurysm, which was successfully treated with coil embolization. The patient had complete resolution of his phantom pain after treatment of the aneurysm, suggesting it was the source of the pain. As a result of successful diagnosis and treatment, the amputation level did not need to be revised and the patient was able to continue his previously high level of function. PMID- 11932861 TI - Predictors of life satisfaction: a spinal cord injury cohort study. AB - OBJECTIVE: To determine unique demographic, medical, perceived health, and handicap predictors of life satisfaction 2 years after spinal cord injury (SCI), as well as the predictors of change in life satisfaction from year 1 to year 2. DESIGN: Prospective predictive study performed by using longitudinal data from 18 Spinal Cord Injury Model Systems. SETTING: University physical medicine and rehabilitation department. PARTICIPANTS: Adults with traumatic onset SCI (N = 940) evaluated at 1 and 2 years' postinjury. INTERVENTIONS: Not applicable. MAIN OUTCOME MEASURES: Satisfaction with Life Scale (SWLS) 2 years post-SCI. PREDICTOR VARIABLES: demographic characteristics, impairment and disability classifications, and 1 year post-SCI measures of life satisfaction (SWLS), medical complications, self-perceived health (Medical Outcomes Study 12-Item Short-Form Health Survey), and extent of handicap (Craig Handicap Assessment and Reporting Technique). RESULTS: The factors uniquely associated with an increased risk of lower self-reported life satisfaction at year 2 post-SCI included being male and unemployed, with poor perceived health, decreased mobility, and decreased social integration. After controlling for year 1 estimates of life satisfaction (ie, examining change in life satisfaction), only mobility and perceived health were uniquely related to life satisfaction 2 years post-SCI. CONCLUSION: Mobility and perceived health appear to be the consistent predictors of life satisfaction at year 2 post-SCI, as well as change in satisfaction from year 1 to year 2. Because both factors are amenable to change, they are reasonable targets of intervention programs. Identifying specific mechanisms of perceived health and mobility associated with life satisfaction should be an important area of continued research. PMID- 11932863 TI - Acute isolated suprascapular nerve palsy limited to the infraspinatus muscle: a case report. AB - Suprascapular nerve palsy resulting in isolated weakness of the infraspinatus muscle is subtle at its onset and must be differentiated from cervical radiculopathy or bone and joint diseases of the shoulder, especially in the presence of pain. The literature suggests an association of cumulative trauma with ganglionic cyst formation and entrapment neuropathy of the suprascapular nerve in the spinoglenoid notch. Here we present clinical, electrodiagnostic, and radiologic features in a young patient who presented with a 2-week history of isolated, painless weakness of the right shoulder. His electromyography showed acute denervation of the right infraspinatus muscle. Magnetic resonance imaging revealed a ganglionic cyst in the spinoglenoid fossa and edema in the infraspinatus muscle consistent with denervation. The patient recovered after removal of the cyst. PMID- 11932864 TI - "EMG disease" with bulbar muscle involvement: a case report. AB - We report the first case of diffuse abnormal insertional activity with bulbar muscle involvement. Electromyography performed 5 months earlier reported multilevel radiculopathy. A repeat electromyography study revealed short trains of positive waves without fibrillation potentials, diffusely present in all tested muscles. Positive waves were also found in the bulbar innervated muscles; these included the trapezius, frontalis, and the orbicularis oculi. This entity is important to recognize because confusion with neuromuscular syndromes can occur. PMID- 11932865 TI - Understanding the pusher behavior of some stroke patients with spatial deficits: a pilot study. AB - OBJECTIVE: To investigate whether pusher behavior (ie, a tendency among stroke patients with spatial deficits to actively push away from the nonparalyzed side and to resist any attempt to hold a more upright posture) affects only the trunk, for which gravitational feedback is given by somesthetic information, or the head as well, whose gravitational information is mainly given by the vestibular system (without vision). DESIGN: Description and measurement of clinical features. SETTING: Rehabilitation center research laboratory. PARTICIPANTS: Eight healthy subjects age matched to 14 patients with left hemiplegia resulting from right hemisphere stroke (3 pushers showing a severe spatial neglect, 11 without pusher behavior). INTERVENTION: All participants were asked to actively maintain an erect posture while sitting for 8 seconds on a rocking, laterally unstable platform. The task was performed with (in light) and without (in darkness) vision. MAIN OUTCOME MEASURES: The number of trials needed to succeed in the task was monitored. In successful trials, head, shoulders, thoracolumbar spine, and pelvis orientation in roll were measured by means of an automated, optical television image processor. RESULTS: Compared with other patients and healthy subjects, the 3 pushers missed many more trials and displayed a contralesional tilt of the pelvis but kept a correct head orientation. This tilt was especially pronounced without vision. Spatial neglect was a key factor, explaining 56% of patients' misorientation behavior with vision and 61% without vision. CONCLUSION: This pilot kinematic analysis shows that pusher behavior does not result from disrupted processing of vestibular information (eg, caused by a lesion involving the vestibular cortex); rather, it results from a high-order disruption in the processing of somesthetic information originating in the left hemibody, which could be graviceptive neglect (extinction). This disruption leads pushers to actively adjust their body posture to a subjective vertical biased to the side opposite the cerebral lesion. PMID- 11932866 TI - Oxygen consumption during ambulation: comparison of using a prosthesis fitted with and without a tele-torsion device. AB - OBJECTIVE: To determine the energy expenditure and subjective comfort rating of walking in transtibial amputee patients using their prosthesis fitted either with or without a tele-torsion device (TT Pylon). DESIGN: Randomized, before-after trial. SETTING: Gait laboratory. PARTICIPANTS: Six men, moderately active, with unilateral transtibial amputation (mean age, 39.5 +/- 9.9y). INTERVENTION: Subjects walked on a motorized treadmill using their prosthesis fitted either with (ProsWith) or without (ProsWithout) a TT Pylon. Trials were repeated with subjects walking at speeds 100%, 130%, and 160% of their "normal" pace. MAIN OUTCOME MEASURES: The energy expended (VO(2)) and subjective comfort rating during each walking trial. RESULTS: VO(2) during walking with the prosthesis fitted with the TT Pylon was 5.4% and 9.1% lower than when using the prosthesis without the TT Pylon, at the speeds 130% and 160% of normal, respectively. Findings at the speed 160% greater than normal were significant (P <.05). Two of the subjects perceived no difference in prosthetic comfort between ProsWith and ProsWithout. The other 4 subjects preferred the TT Pylon at all speeds. CONCLUSION: Use of a TT Pylon can significantly reduce the energy expenditure of walking at speeds above normal. PMID- 11932867 TI - Reliability of muscle strength testing quantified by the intraclass correlation coefficient. PMID- 11932868 TI - Mobility scales for lower limb-prosthetic patient: the locomotor capabilities index. PMID- 11932870 TI - Need for circumspection in prescribing tumor necrosis factor inhibitors and other biologic response modifiers. PMID- 11932871 TI - Alternative medicine and The Arthritis Foundation. PMID- 11932872 TI - Assessment of fatigue in patients with ankylosing spondylitis: a psychometric analysis. AB - OBJECTIVES: To investigate whether the single-item fatigue question of the Bath Ankylosing Spondylitis Disease Activity Index (BASDAI) and the Multidimensional Fatigue Inventory (MFI) are appropriate instruments to measure fatigue in ankylosing spondylitis (AS); to identify factors that influence fatigue in AS; and to assess how fatigue in all its aspects is associated with quality of life in AS. METHODS: A total of 812 patients with AS were included. Patients completed questionnaires on disease activity (BASDAI), functional ability (Bath Ankylosing Spondylitis Functional Index [BASFI]), global well-being (Bath Ankylosing Spondylitis Global Score [BAS-G]), overall perceived health (EuroQoL visual analog scale), and quality of life (Ankylosing Spondylitis Quality of Life questionnaire, and Short Form 36 [SF-36]). Patients were dichotomized into a F+ group (fatigue = major symptom) if the BASDAI fatigue score was > 5.0 and a F- group (fatigue = minor symptom) if the fatigue score was < 5.0. Reproducibility was assessed with intraclass correlation coefficients, and responsiveness was calculated according to 3 different methods. Logistic regression analysis was used to determine which factors were associated with fatigue. Multiple regression analysis was used to investigate whether fatigue contributes in explaining quality of life. RESULTS: Fifty-three percent of the patients were assigned to the F+ group. They scored significantly worse compared with the F- group with respect to each dimension of the MFI and to all other questionnaires studied (all P < 0.001). The BASDAI fatigue question, as well as each separate dimension of the MFI, showed moderate to good reproducibility (0.57-0.75) and responsiveness (0.23-0.96). Fatigue was, in the opinion of the patients, highly associated with pain (70% of patients) and stiffness (54% of patients). Logistic regression analysis showed that scores on BASDAI, BASFI, BAS-G, and mental health status (SF 36) were independently associated with fatigue (R(2) = 0.52). Multiple regression analysis showed that scores on the BASDAI fatigue question were significantly associated with quality of life. With the 5 MFI dimensions as explanatory variables, different aspects of fatigue were associated with different domains of quality of life. CONCLUSIONS: Fatigue as a major symptom of AS can effectively be measured with either a single-item question on the intensity of fatigue or with the MFI. The MFI, however, provides more insight into specific dimensions of fatigue. Fatigue appears to be associated with the level of disease activity, functional ability, global well-being, and mental health status. In addition, fatigue negatively influences different aspects of quality of life. PMID- 11932873 TI - Experience with etanercept in an academic medical center: are infection rates increased? AB - OBJECTIVES: There is little established information regarding the safety of antitumor necrosis factor therapies used outside the setting of clinical trials. This study evaluated the long-term safety and tolerability of open-label use of etanercept when used to treat patients with a variety of systemic rheumatic diseases. Reduction of concomitant corticosteroid and disease-modifying antirheumatic drug was also assessed. METHODS: Retrospective medical record review of 180 patients who were started on etanercept between December 1998 and April 2000 at an academic medical center. RESULTS: Most patients (81%) remained on therapy for longer than 6 months, and a significant number (43%) of patients for longer than 12 months. Etanercept was prescribed for rheumatoid arthritis (RA) in 144 patients and for diseases other than RA, including ankylosing spondylitis, psoriatic arthritis, and polymyositis, in 36 patients. Fifty-six percent of patients taking corticosteroids were able to reduce their dose and 51% of patients were able to taper their methotrexate dosages. Forty-three patients (26%) discontinued etanercept. Reasons for discontinuing therapy included serious adverse events (2.9%), of which infection was most common. These included a psoas abscess secondary to Mycobacterium avium-intracellulare, septic wrist, bacteremia, and septic total hip replacement. Two deaths associated with infection were seen. CONCLUSIONS: The majority of the studied patients tolerated etanercept for longer than 6 months. Many of these patients were able to subsequently taper or even discontinue corticosteroid and methotrexate therapy. Serious infections occurred in this patient population. Our results underscore the value of long-term observation under the conditions of clinical practice beyond controlled clinical trials. PMID- 11932874 TI - The effects of progressive strength training and aerobic exercise on muscle strength and cardiovascular fitness in women with fibromyalgia: a pilot study. AB - OBJECTIVE: To determine the safety, feasibility and consequences of a program of progressive strength training and cardiovascular exercise in women with fibromyalgia syndrome (FMS). METHODS: Fifteen women with confirmed FMS were monitored for injury and exercise compliance, and assessed for muscle strength (1 repetition maximum technique), cardiovascular endurance (6-minute walk test), and functional status (Fibromyalgia Impact Questionnaire [FIQ]) before and after a 20 week exercise intervention. RESULTS: Zero injuries and an 81% compliance rate occurred during training. Improvement was seen in muscle strength of the lower (191 +/- 75 to 265 +/- 67 pounds; P < 0.001) and upper (61 +/- 18 to 76 +/- 18 pounds; P < 0.001) body, 6-minute walk distance (530 +/- 80 to 629 +/- 74 meters; P < 0.001), and in FIQ score (44 +/- 9 to 32 +/- 14; P < 0.01). CONCLUSION: A program of progressive strength training and cardiovascular exercise can be safe, well tolerated, and effective at improving muscle strength, cardiovascular endurance and functional status in women with FMS without exacerbating symptoms. This program may also contribute to a reduction in the severity of several symptoms. PMID- 11932875 TI - A strong initial systemic inflammatory response is associated with higher corticosteroid requirements and longer duration of therapy in patients with giant cell arteritis. AB - OBJECTIVE: To assess whether the intensity of the initial systemic inflammatory response is able to predict response to therapy in patients with giant cell arteritis (GCA). METHODS: Retrospective review of 75 patients (49 women and 26 men) with biopsy-proven GCA who had regular followup and were treated according to uniform criteria. Four parameters were used to evaluate the baseline inflammatory response at diagnosis: fever, weight loss, erythrocyte sedimentation rate > or = 85 mm/hour, and hemoglobin < 110 gm/liter. Patients were considered to have a weak inflammatory response if they had 2 or fewer inflammatory parameters (group 1) and a strong inflammatory response if 3 or 4 parameters were present (group 2). Time required to achieve a maintenance dose of less than 10 mg prednisone/day was recorded and analyzed by the Kaplan-Meier survival analysis method. Tumor necrosis factor alpha (TNFalpha) and interleukin 6 (IL-6) serum levels were also determined in 62 patients and 15 controls. RESULTS: Forty patients had a weak (group 1) and 35 had a strong (group 2) initial inflammatory response. Patients in group 2 had significantly higher levels of circulating TNFalpha (31.9 +/- 16.8 versus 22.3 +/- 9 pg/ml; P = 0.01) and IL-6 (28.2 +/- 17.4 versus 16.6 +/- 13 pg/ml; P = 0.004) than patients in group 1. In group 1, 50% of patients required a median of 40 weeks (95% CI 37-43) to reach a maintenance dose of <10 mg, whereas in group 2 a median of 62 weeks (95% CI 42 82) was necessary (P = 0.0062). Patients in group 2 experienced more flares than patients in group 1 (P = 0.01) and received higher cumulative steroid doses (8.974 +/- 3.939 gm versus 6.893 +/- 3.075 gm; P = 0.01). CONCLUSION: GCA patients with a strong initial systemic inflammatory reaction have more elevated circulating levels of IL-6 and TNFalpha, have higher and more prolonged corticosteroid requirements, and experience more disease flares during corticosteroid therapy than patients with a weak systemic acute phase response. PMID- 11932876 TI - Role of initial NSAID choice and patient risk factors in the prevention of NSAID gastropathy: a decision analysis. AB - OBJECTIVE: To investigate the role of initial nonsteroidal antiinflammatory drug (NSAID) choice in the prevention of NSAID gastropathy, based on relative clinical and economic effects. METHODS: To mimic clinical practice, a symptom-driven decision analytic model was constructed to compare 2 treatment strategies for long-term users of NSAIDs over a 1-year period: Strategy 1-generic NSAID used initially, and safer, more expensive NSAID reserved for treatment failures due to symptomatic gastropathy; and Strategy 2-safer, more expensive NSAID used in all instances. The only distinction between the strategies was the choice of initial NSAID. NSAIDs differed in gastrointestinal safety profiles and acquisition costs. The use and impact of antisecretory medications were included in the model. Because published data on patients' ulcer risk and relative NSAID safety show considerable variability, sensitivity analyses were used to evaluate the key clinical outcomes and costs. RESULT: For patients without risk factors for NSAID ulcers (average risk), the model estimated that the strategy restricting use of the safer NSAID resulted in more symptomatic ulcers (Strategy 1, 2.58; Strategy 2, 0.73) and ulcer-related complications (Strategy 1, 1.18; Strategy 2, 0.23) per 100 patient years. The restricted strategy led to a significantly lower cost per patient treated (Strategy 1, $239; Strategy 2, $831 per year). In the principal analysis, the incremental costs to prevent symptomatic and complicated ulcers were $31,900 and $56,700, respectively. The estimated incremental cost per ulcer avoided was sensitive to the relative protection provided by the safer NSAID and fell dramatically as the patients' ulcer risk was increased above average risk. CONCLUSION: Unrestricted use of NSAIDs that reduce the risk of symptomatic ulcers has the potential to produce important clinical benefits at incremental cost. The impressive impact of ulcer risk on the incremental cost per ulcer prevented warrants increased attention to risk factor identification when NSAIDs are prescribed. PMID- 11932877 TI - Joint protection and home hand exercises improve hand function in patients with hand osteoarthritis: a randomized controlled trial. AB - OBJECTIVE: To determine the effect of joint protection and home exercises on hand function of patients with hand osteoarthritis (OA). METHODS: Randomized, controlled, 3-month trial with a blinded assessor. Primary outcome parameter was grip strength; secondary parameters were Health Assessment Questionnaire and visual analog scales (VAS) for pain and global hand function. Forty patients with hand OA were randomly assigned to 2 groups: One group received instruction for joint protection and home hand exercises (JPE group), the control group received an information session about hand OA. RESULTS: Grip strength improved by 25% in the JPE group (right hand, P < 0.0001; left hand, P = 0.0005), but not in the control group. Global hand function (by VAS) improved in a larger proportion (65%) of patients in the JPE group (P < 0.05). CONCLUSIONS: Joint protection and hand home exercises, easily administered and readily acceptable interventions, were found to increase grip strength and global hand function. PMID- 11932878 TI - Structural effect of avocado/soybean unsaponifiables on joint space loss in osteoarthritis of the hip. AB - OBJECTIVE: To evaluate the structural effect of avocado/soybean unsaponifiables (ASU) in the treatment of patients with symptomatic osteoarthritis (OA) of the hip. METHODS: Patients with regular painful primary OA of the hip (European League Against Rheumatism 1980 criteria) and a joint space still > or = 1 mm (Kellgren grade 1 to 3, assessed by an independent observer prior to inclusion) entered a prospective, multicenter, randomized, parallel group, double-blind, placebo-controlled trial of 2 years duration. Patients had at least a 6-month history of regular pain and an algofunctional index (AFI) > or = 4. The primary assessment criterion was a decrease of the joint space width (JSW) on plain anteroposterior radiographs of the pelvis performed in standing position, measured at the narrowest points by 2 independent readers, previously tested and selected and blinded to both the treatment and the time sequence. Secondary criteria were standard clinical outcome measurements (AFI, pain on a visual analog scale, consumption of nonsteroidal antiinflammatory drugs and patient's and investigator's global assessments). RESULTS: One hundred sixty-three patients were included: 102 men and 61 women (mean age 63.2 +/- 8.7 years). A total of 108 patients (72 men and 36 women; mean age 64 +/- 7.9 years) were radiologically evaluable at 23.7 +/- 2.6 months (ASU group; n = 55) and 23.7 +/- 3.2 months (placebo group; n = 53). Overall comparison of the evolution of JSW showed no difference between the ASU and placebo groups, from 2.35 +/- 0.93 to 1.87 +/- 1.10 mm and from 2.5 +/- 0.94 to 1.9 +/- 1.33 mm, respectively (intergroup P value at end point = 0.9). When patients were divided into 2 subgroups according to the median value of the baseline JSW (2.45 mm), the joint space loss in the most severely affected subgroup of patients (baseline JSW < or = median) was significantly greater in the placebo group than in the ASU group: from 1.69 +/- 0.58 to 0.84 +/- 0.77 mm (-0.86 +/- 0.62 mm) and from 1.66 +/- 0.42 to 1.22 +/- 0.7 mm (-0.43 +/- 0.51 mm), respectively (P < 0.01). The JSW decrease was identical, with no difference in ASU and placebo groups, in the less severely affected subgroup of patients (baseline JSW > median). Clinical parameters in the 2 groups did not differ significantly throughout the study. CONCLUSION: This pilot randomized, double-blind, placebo-controlled trial failed to demonstrate a structural effect of ASU in hip OA. However, in a post-hoc analysis, ASU significantly reduced the progression of joint space loss as compared with placebo in the subgroup of patients with advanced joint space narrowing. These results suggest that ASU could have a structural effect but require confirmation in a larger placebo-controlled study in hip OA. PMID- 11932879 TI - Step-down approach using either cyclosporin A or methotrexate as maintenance therapy in early rheumatoid arthritis. AB - OBJECTIVE: To evaluate the feasibility and outcome of the step-down approach using either cyclosporin A (CSA) or methotrexate (MTX) as maintenance therapy following 6 months treatment with these 2 agents in combination in early, nonerosive rheumatoid arthritis (RA). METHODS: Fifty-seven patients younger than 65 years with early, nonerosive RA were first treated with CSA and MTX in combination for 6 months. They were then randomly stepped down to single-agent maintenance treatment for another 18 months. Safety, clinical efficacy, survival on treatment, and radiographic progression were evaluated. RESULTS: When being treated with combination therapy, 7 of the 57 patients (12.3%) withdrew because of adverse events. Of the remaining 50 patients, 42 (84.0%) were American College of Rheumatology (ACR) 20% responders, 30 (60.0%) were ACR 50% responders, and 23 (46.0%) were ACR 70% responders. At month 6, 22 patients were randomized to CSA and 27 to MTX. During this trial period, the treatment was discontinued by 16 patients taking CSA (mainly because of loss of efficacy) and by 4 taking MTX. At month 24, the probability (+/- SEM) of survival on treatment was 0.273 +/- 0.09 for CSA and 0.852 +/- 0.07 for MTX. Of the 6 CSA patients who completed the trial, 4 (66.7%) were ACR 20% responders, and 3 (50%) were both ACR 50% and ACR 70% responders. Of the 23 completers in the MTX arm, 21 (91.3%) were ACR 20% responders, 18 (78.3%) were ACR 50%, and 10 (43.5%) were ACR 70% responders. The treatment was not responsible for severe adverse events. Radiography showed a slow progression in the damage score and number of eroded joints in both treatment groups. CONCLUSION: Stepping down to single agent maintenance therapy following 6 months of combination treatment with CSA and MTX in early RA was only successful with MTX. Because this treatment did not prevent some radiographic progression, other approaches (e.g., step-up approach) may be more appropriate in early RA. PMID- 11932880 TI - The number of deformed joints as a surrogate measure of damage in rheumatoid arthritis. AB - OBJECTIVE: To evaluate the reliability and validity of the number of deformed joints (NDJ) as a surrogate measure of joint damage in rheumatoid arthritis (RA). METHODS: We tested interrater reliability and validity in determining the NDJ as a surrogate for joint damage in consecutive patients with RA. We rated each of 48 joints as normal or abnormal in terms of alignment and range of motion, and expressed the results as the total number of deformed joints. We compared the NDJ with the severity of damage on a plain radiograph of the hands, scored using Sharp's technique, as the gold standard measure of joint damage. We also compared the correlation between the NDJ and radiographic joint damage, on the one hand, and disease duration, performance-based measures of physical function, and the self-reported level of disability. RESULTS: The interrater reliability of the NDJ was excellent, with an intraclass correlation among four examiners of 0.94. To assess validity of the NDJ, we studied 273 RA patients from 5 clinical settings. Their average NDJ was 14 (range 0-43), and their average Sharp's score for joint space narrowing and erosions combined was 106 (range 4-309). The NDJ and the total Sharp's score were highly correlated (r = 0.83). Both measures were correlated to a similar degree with disease duration (r = 0.51 for each measure), grip strength (r = -0.49 for NDJ, and r = -0.51 for Sharp's score), walking velocity (r = -0.44 for NDJ, and r = -0.45 for Sharp's score), the timed button test (r = -0.62 for NDJ, and r = -0.57 for Sharp's score), and the Modified Health Assessment Questionnaire (r = 0.38 for NDJ, and r = 0.38 for Sharp's score). Both the Sharp's score and the NDJ worsened significantly in 38 patients for whom 1-2 year followup data were available. CONCLUSION: The NDJ is reliable and is strongly associated with the standard measure of joint damage in RA. Because it is easily performed in a clinical setting, it could be used as an economical surrogate of joint damage in studies of the long-term outcome of RA. PMID- 11932881 TI - Design and quality considerations for randomized controlled trials in systemic sclerosis. AB - OBJECTIVE: To appraise systematically randomized controlled trials (RCTs) on systemic sclerosis (SSc) in order to determine whether the parameter of study design and its quality may influence the reporting of efficacy for tested interventions. METHODS: Seventy RCTs were analyzed (1965-2000) in terms of design, patient characteristics, outcomes, and reported results. RESULTS: Median sample size was 28 patients. Fifty-nine trials were double blind, but only 16 mentioned the randomization mode and only 7 described allocation concealment. There was sufficient information on withdrawals in 37 trials. Larger trials with longer followup scored higher on quality characteristics, but had higher withdrawal rates. Only 8 trials had a followup of more than 1 year. Significant efficacy was less likely to be reported in double-blind studies (P = 0.029) and in studies with larger rates of withdrawal (P = 0.032). Specification of the following parameters improved over time: power calculations (P = 0.0003), outcomes (P = 0.001), and sample size per arm (P = 0.011). CONCLUSIONS: Several aspects of the quality of design and conduct of SSc RCTs can be improved. Adequately powered trials with longer followup and clear outcomes are needed. PMID- 11932882 TI - The effect of graded resistance exercise on fibromyalgia symptoms and muscle bioenergetics: a pilot study. PMID- 11932883 TI - Combination cytokine therapy: the next generation of rheumatoid arthritis therapy? PMID- 11932884 TI - American College of Rheumatology Basic Research Conference: Genetics and genomics in rheumatic disease. PMID- 11932885 TI - Splinting the juvenile arthritic wrist: a clinical observation. PMID- 11932886 TI - Cryptococcal arthritis, tendinitis, tenosynovitis, and carpal tunnel syndrome: report of a case and review of the literature. PMID- 11932887 TI - Are you better? Comment on the article by Beaton et al. PMID- 11932888 TI - Vesalius, also a rheumatologist? PMID- 11932890 TI - Jonathan Evans Rhoads, M.D.: in memoriam 1907-2002. PMID- 11932891 TI - Multicentric mammary carcinoma: evidence of monoclonal proliferation. AB - BACKGROUND: Both the widespread use of screening mammography and emphasis on breast conservation have raised many questions regarding the clinical and therapeutic management of multicentric mammary carcinoma (MMC). MMC has been postulated to be either a clonal proliferation of a single mammary carcinoma or multiple independent synchronous primary tumors in the same breast. The goal of the current study was to evaluate the histologic features and immunohistochemical profile of MMC. We also compared the clinical outcomes of the patients in the current study with stage-matched and treatment-matched groups of patients with unicentric mammary carcinoma. METHODS: The authors studied 32 patients with T1 T2, N0-1, M0 multicentric invasive mammary carcinomas diagnosed between 1983-1988 and treated at The University of Texas M. D. Anderson Cancer Center. The histologic features of each tumor (including tumor type, nuclear grade, presence of in situ carcinoma, pattern of in situ carcinoma, and lymphovascular invasion) were evaluated. The authors performed immunohistochemical analysis of estrogen receptor (ER), progesterone receptor (PR), HER-2/neu, and Ki-67 in 25 cases, including 14 from which > 1 tumor was available to perform comparative immunohistochemical analysis. The clinical parameters of each case were compared with those of the unicentric breast carcinoma controls. RESULTS: The median age of the patients with MMC was 45 years (range, 28-69 years). Twelve patients had a family history of breast carcinoma (37.5%). The maximum tumor dimension ranged from 0.2-3.2 cm in the index lesion (median, 2.0 cm) and 0.1-2.5 cm in the second lesion (median, 0.9 cm). Twelve patients were clinically classified as having Stage I disease and 20 patients were considered to have Stage II disease at the time of presentation. Follow-up data were available for all the patients and follow-up ranged from 4.5-16 years (median, 6 years). The disease-free survival was 84% at 5 years and 73% at 10 years in the MMC patients and 78% and 70%, respectively, in patients with unicentric breast carcinoma (P = 0.4368). Histologically, 24 of the multicentric tumors were found to be infiltrating ductal tumors and 8 were found to be infiltrating lobular carcinomas. The nipple was involved in 10 cases. The histology of the multicentric invasive tumor was nearly identical in 31 cases (97%). Approximately 72% of the cases had in situ carcinoma in both tumors and 44% had lymphovascular invasion. Comparative immunohistochemical analysis of separate tumors was equivalent with regard to ER, PR, and HER-2/neu. The quantitative immunohistochemical staining for the proliferative marker Ki-67 differed between tumors in two cases. CONCLUSIONS: The near-identical morphologic and immunohistochemical patterns in the MMC cases in the current study support the hypothesis that early-stage synchronous tumors are a clonal proliferation of a single mammary carcinoma. Furthermore, the results of the current study support evaluating prognostic markers in only one tumor per MMC patient. There was no appreciable difference in the disease-free survival of patients with unicentric and multicentric breast carcinoma. PMID- 11932892 TI - The value of breast lumpectomy margin assessment as a predictor of residual tumor burden in ductal carcinoma in situ of the breast. AB - BACKGROUND: Margin width is considered the most important risk factor for local recurrence in ductal carcinoma in situ (DCIS) of the breast. The purpose of this report is to assess the predictive utility of lumpectomy specimen margin assessment for the presence and extent of residual DCIS. METHODS: Specimens from 253 DCIS cases with lumpectomy and reexcision were studied to determine to the probability of residual DCIS on reexcision. The probability of residual tumor was evaluated with respect to tumor size, margin status, nuclear grade, presence of necrosis, patient age, and the extent of specimen processing (number of sections/volume tissue). Lesions were grouped by size: less than or equal to 2 mm, greater than 2-15 mm, greater than 15-40 mm, or greater than 40 mm. Margin width was recorded as the distance of DCIS to the closest specimen edge or, for positive margins, scored as: extensive (margin involvement in > or =8 sections or >4 low-power fields [LPFs]), moderate (5-7 sections or 2-4 LPFs), minimal (2-4 sections or 1 LPF), or focal (1 section, single focus). The amount of residual tumor was graded by maximum dimension on a semiquantitative basis. RESULTS: Initial excision margin significantly predicted for the presence of residual tumor on reexcision. Residual tumor was found on reexcision in 85% of extensively positive, 68% of moderately positive, 46% of minimally positive, 30% of focally positive, 41% of greater than 0-1 mm, 31% of greater than 1-2 mm, and 0% of greater than 2 mm margins (P < 0.0001). On univariate analysis, margin width and lesion size of initial excision specimens significantly predicted for the presence of residual DCIS on reexcision. Age, grade, necrosis, and extent of specimen processing were not significant prognostic factors. On multivariate analysis, both initial margin width (P < 0.0001) and lesion size (P = 0.02) significantly predicted for residual DCIS. As for amount of residual tumor, margin width and initial lesion dimension both significantly predicted for medium to large residuum, whereas age 45 years or younger was of borderline significance on univariate analysis. On multivariate analysis, margin width and lesion size on initial excision both remained significant predictors of larger volume residual tumor. CONCLUSIONS: The margin status of a DCIS lumpectomy specimen is the most important predictive factor for both the presence and amount of residual disease. PMID- 11932893 TI - A Phase II study of paclitaxel by 24-hour infusion and ifosfamide in anthracycline-resistant metastatic breast carcinoma. AB - BACKGROUND: A Phase II study was performed to investigate the efficacy and tolerability of paclitaxel and ifosfamide chemotherapy for the treatment of anthracycline-resistant metastatic breast carcinoma (MBC). METHODS: Recurrent or progressed MBC within 12 months after anthracycline-based chemotherapy was defined as anthracycline-resistant. A 24-hour infusion of paclitaxel (175 mg/m(2)) on Day 1 and subsequent infusions of ifosfamide (1.8 g/m(2)/day) with mesna (360 mg/m(2)/day) on Days 2- 4, were performed every 3 weeks. Twenty-one patients were eligible for toxicity analysis. Response rate and survival duration were evaluated in 21 patients. Frontline chemotherapy was the FAC (5 fluorouracil, doxorubicin, cyclophosphamide) regimen in all patients. RESULTS: Objective response was found in 9 patients (42.9%), including complete response in 3 (13.4%). Median response duration and median survival duration were 10 months (range, 2-24+) and 19+ months (range, 2-32+), respectively. Sixteen (76%) experienced Grade 3/4 leukopenia controllable with granulocyte macrophage colony stimulating factor. Other significant toxicities were peripheral neuropathy (n = 3), mucositis (n = 2), and liver dysfunction (n = 1). However, there was no chemotherapy-related death. CONCLUSIONS: Paclitaxel by 24-hour infusion combined with ifosfamide is efficacious in the treatment of anthracycline-resistant MBC with tolerable toxicity. Further trials verifying the result of the authors' study are warranted. PMID- 11932894 TI - Efficacy and toxicity of adjuvant chemotherapy in elderly patients with colon carcinoma: a 10-year experience of the Geisinger Medical Center. AB - BACKGROUND: Although the benefit from adjuvant chemotherapy has been established clearly in patients with Stage III colon carcinoma, the degree to which elderly patients with colon carcinoma can tolerate such therapy generally has remained unknown. METHODS: The authors reviewed all patients in their Tumor Registry with Stage II and Stage III adenocarcinoma of the colon who underwent potentially curative resection for their disease at the Geisinger Medical Center between January 1990 and September 2000. One hundred twenty patients underwent complete resection of their colon carcinoma and received 5-fluorouracil-based (5-FU) adjuvant chemotherapy. RESULTS: The 5-year disease free survival rate for patients age > or =65 years (Group A) was 70% compared with 56% for patients age < 65 years (Group B) (P = 0.085). The 5-year overall survival rate for patients in Group B was 77% compared with 62% for the patients in Group A (P = 0.143). In a Cox regression model, age was not a predictor of disease free survival (P = 0.633) or overall survival (P = 0.900) when it was analyzed as a continuous variable. Only 19 patients were age > 75 years, and the disease free and overall survival rates for this group were similar but were underpowered compared with the rates for the patients ages between 65-75 years. When gender and disease stage were included in the model, age remained a nonsignificant variable (P = 0.400 for disease free survival; P = 0.615 for overall survival). Nine of 56 patients in Group A (16%) experienced Grade 3-4 toxicity compared with 14 of 64 patients in Group B (22%) (P = 0.420). The lack of a correlation between toxicity and age was maintained after controlling for disease stage and patient gender (P = 0.343). There were no correlations between preoperative carcinoembryonic antigen level, tumor grade, or lymph node involvement and patient age (P = 0.258, P = 0.256, and P = 0.519, respectively). CONCLUSIONS: Elderly patients with Stage II and Stage III colon carcinoma benefit from 5-FU-based adjuvant therapy without a significant increase in toxicity compared with their younger counterparts. Adjuvant chemotherapy should be presented to elderly patients with high-risk, resected colon carcinoma. The data regarding age cannot be generalized to patients age > 75 years. PMID- 11932895 TI - Expression of UDP-N-acetyl-alpha-D-galactosamine-polypeptide galNAc N acetylgalactosaminyl transferase-3 in relation to differentiation and prognosis in patients with colorectal carcinoma. AB - BACKGROUND: Tumor development usually is accompanied by alterations of O glycosylation. Initial glycosylation of mucin-type, O-linked proteins is catalyzed by one of the UDP-GalNAc-polypeptide N-acetyl-galactosaminyl transferases, such as GalNAc-T3, which is expressed in adenocarcinoma cells. The authors investigated whether such expression influenced tumor differentiation or prognosis in patients with colorectal carcinoma. METHODS: The expression of GalNAc-T3 was evaluated immunohistochemically in 106 paraffin embedded samples from surgically resected colorectal carcinomas and was related to patient and tumor characteristics. Western blot analysis was performed on seven samples of frozen tissue. RESULTS: Strong tumor expression of GalNAc-T3 predicted 5-year survival in patients with colorectal carcinoma (67.2% vs. 43.6% for weak expression; P = 0.017). GalNAc-T3 expression was not associated with age, gender, tumor size, tumor location, or disease stage but was related to histologic differentiation (P = 0.049) and depth of invasion (P = 0.031). Univariate analysis showed that strong GalNAc-T3 expression significantly enhanced the likelihood of survival. Multivariate Cox survival analysis identified enzyme expression as an independent prognostic factor that was second only to TNM stage. CONCLUSIONS: GalNAc-T3 expression is a novel and useful indicator of tumor differentiation, disease aggressiveness, and prognosis in patients with colorectal carcinoma. PMID- 11932896 TI - Prognostic significance of serum thymidine phosphorylase concentration in esophageal squamous cell carcinoma. AB - BACKGROUND: Thymidine phosphorylase (dThdPase), which also is referred to as platelet-derived endothelial cell growth factor, is a potent inducer of angiogenesis in malignant tumors. Increased dThdPase expression and activity have been found to be associated with poor prognosis in various solid tumor tissues. Because very little was known about the significance of serum dThdPase concentration (S-dThdPase), the objective of this study was to analyze the clinicopathologic significance of S-dThdPase in the patients with esophageal squamous cell carcinoma. METHODS: The S-dThdPase was measured by enzyme-linked immunosorbent assay in 77 healthy controls and 153 patients with primary esophageal squamous cell carcinoma. A total of 80 patients underwent surgery alone; 46 patients received chemoradiotherapy alone; 17 patients received chemoradiotherapy followed by surgery; and 10 patients did not receive any treatment. Thymidine phosphorylase expression in esophageal carcinoma tissues was examined by immunohistochemistry. The clinicopathologic value and prognostic value of S-dThdPase was determined in 80 patients treated by surgery alone. RESULTS: The S-dThdPase is significantly higher in patients with esophageal carcinoma than in healthy controls (30.8 +/- 31.8 ng/mL vs. 13.8 +/- 7.6 ng/mL; P < 0.001). Statistically significant differences in S-dThdPases were observed depending on tumor size (P < 0.01) and tumor depth (P < 0.01). A S-dThdPase of more than 29.0 ng/mL (which represented the mean plus 2 standard deviation of the concentration in healthy controls) was associated with dThdPase expression (P = 0.022), poor response (P = 0.022), and poor survival (P < 0.01). Because S dThdPase was associated with tumor depth, S-dThdPase was not an independent prognostic factor (P = 0.095). CONCLUSIONS: A high S-dThdPase is associated with depth of tumor invasion and poor response to treatment. PMID- 11932897 TI - Expression and prognostic significance of PTEN product protein in patients with esophageal squamous cell carcinoma. AB - BACKGROUND: PTEN is a candidate tumor-suppressor gene in a variety of malignant tumors. The prognostic importance of PTEN product protein (PTEN) and its correlation with clinicopathologic characteristics have yet to be delineated in patients with esophageal carcinoma. METHODS: Specimens from 97 patients with esophageal squamous cell carcinoma were used for the immunohistochemical evaluation of PTEN expression. RESULTS: PTEN expression was detected in the nucleus in 48 specimens (49.5%). There were statistically significant correlations between nuclear PTEN expression and macroscopic tumor classification, T stage, and American Joint Committee on Cancer (AJCC) stage (P < 0.01), indicating that PTEN expression was down-regulated by advancement of the disease process. There were no statistically significant correlations between nuclear PTEN expression and the intensity and extent of cytoplasmic PTEN expression. The 10-year overall survival rate was significantly better in patients with positive nuclear PTEN expression (n = 48 patients) compared with the rate in patients with negative nuclear PTEN expression (n = 49 patients; P < 0.01). The results of a multivariate analysis of factors that were prognostic for survival showed that AJCC stage (P < 0.05; relative risk, 2.038) and negative nuclear PTEN expression (P < 0.05; relative risk, 1.825) were significant factors indicative of poor survival. CONCLUSIONS: Nuclear PTEN expression may be a favorable biologic marker and a useful prognostic indicator in patients with esophageal squamous cell carcinoma. PMID- 11932898 TI - Performance status rather than age is the key prognostic factor in second-line treatment of elderly patients with epithelial ovarian carcinoma. AB - BACKGROUND: Intravenous cytostatic agents as second-line treatment of epithelial ovarian carcinoma have been withheld from many elderly patients because of fear of toxicity. The purpose of the study is to compare the toxicity and efficacy between elderly (older than 65 years of age) and younger (younger than 65 years of age) patients receiving intravenous second-line treatment of epithelial ovarian carcinoma. METHODS: This study was a retrospective analysis of 286 consecutive patients with primary epithelial ovarian carcinoma. Inclusion criteria included histopathologically documented International Federation of Gynecology and Obstetrics (FIGO) Stage IC-IV epithelial ovarian carcinoma; first line treatment with paclitaxel and a platinum analog; intravenous second-line treatment with topotecan 1.0 mg/m(2)/day for 5 days, every 3 weeks or paclitaxel (175 mg/m(2)) and carboplatin (AUC 5), every 3 weeks. RESULTS: One hundred two patients fulfilled the inclusion criteria receiving topotecan (n = 57) or paclitaxel-carboplatin (n = 45) because of refractory or recurrent disease. The patients' age at start of second-line treatment in the younger (n = 68) and the elderly (n = 34) group were median 54.0 years (range, 34.7-64.3) and 69.5 years (range, 65.1-77.2), respectively. In the patient group aged younger than 65 years, initial performance status was more favorable than in patients aged older than 65 years (P = 0.007) whereas the groups were similar in relation to other potential prognostic factors (P > 0.05). For patients aged younger than 65 years, the overall response rate of 50% (95% confidence interval [CI], 37-63%) was similar to the response rate of 44% (95% CI, 27-62%) in patients aged older than 65 years (P = 0.29). The overall survival from the first day of second-line treatment in patients aged younger and older than 65 years were median 13.3+ months (range, 1.2-38.3+) and 11.8+ months (range, 2.0-41.0+), respectively (P = 0.25). In a multivariate Cox analysis, performance status at time of first-line treatment (0 vs.1-2; P = 0.013; hazard ratio [HR], 2.12), performance status at time of second-line treatment (0 vs. 1-2; P = 0.004; HR, 2.47), and response to second-line treatment (CR + PR vs. NC + PD; P < 0.001; HR, 4.38) were found to be independent significant factors for overall survival whereas age (younger than 65 years vs. older than 65 years) yielded no independent information (P = 0.90). No differences in the rate of postponement of treatment, neutropenia Grade 4, thrombocytopenia Grade 3-4, nor hypersensitivity reaction to either cytostatic agent between older and younger patients were noticed (P > 0.05). CONCLUSIONS: Modality of second-line treatment of epithelial ovarian carcinoma should be determined more by assessment of performance status than age per se. Second-line treatment with topotecan or paclitaxel-carboplatin can be safely administered in the aged. PMID- 11932899 TI - Prognostic value of p53 mutations, bax, and spontaneous apoptosis in maxillary sinus squamous cell carcinoma. AB - BACKGROUND: Many researchers have attempted to correlate p53 mutation and spontaneous apoptosis with the effectiveness of radiochemotherapy and with prognosis in several malignancies. METHODS: The current study group consisted of 70 Japanese patients with maxillary sinus squamous cell carcinoma (SCC). Fifty seven patients were treated with radiochemotherapy followed by total or partial maxillectomy, and the remaining 13 patients were treated with radiotherapy alone. Tumor biopsy specimens at pretreatment status were examined for apoptosis-related proteins such as p53 protein, Fas, bax, bcl-x, and apoptosis using immunohistologic methods. The proportion of apoptotic cells labeled by single stranded DNA antibody was expressed as an apoptotic index (AI). p53 mutations at exons 5 through 8 were analyzed by direct sequence on polymerase chain reaction amplified products obtained from laser microdissected tissues. The effectiveness of radiochemotherapy was investigated histologically on surgically dissected specimens. RESULTS: p53 mutations were identified in 20 (29%) of 70 patients. p53 protein was overexpressed in 39 patients (56%), Fas in 20 patients (29%), bax in 40 patients (57%), and bcl-x in 33 patients (47%). Overexpression of bax was associated with negativity of bcl-x (P = 0.015) and with high AI (P = 0.024). Low AI and/or p53 mutation in the pretreatment tissues correlated with low histologic effectiveness of radiochemotherapy (P = 0.048, P = 0.019, respectively). Kaplan Meier analysis as well as univariate analysis using the Cox proportional hazards model showed that low histologic effectiveness of radiochemotherapy (P = 0.0281, P = 0.0284, respectively), p53 mutations (P = 0.0095, P = 0.0187, respectively), negativity of bax (P = 0.0069, P = 0.0191, respectively), and low AI (P = 0.0134, P = 0.0407, respectively) were significantly related to worse disease-free survival. Multivariate analysis showed AI as an independent factor predicting for disease-free survival (P = 0.0455). CONCLUSIONS: The p53 mutations, expression of bax, and levels of spontaneous apoptosis have prognostic value in maxillary sinus SCC; AI especially is an independent factor for disease-free survival. A high level of spontaneous apoptosis induced by overexpression of bax may increase sensitivity of radiochemotherapy resulting in good prognosis, while p53 mutation may lead to resistance against radiochemotherapy, resulting in poor prognosis. PMID- 11932900 TI - Detection of recurrent or persistent nasopharyngeal carcinomas after radiotherapy with technetium-99m methoxyisobutylisonitrile single photon emission computed tomography and computed tomography: comparison with 18-fluoro-2-deoxyglucose positron emission tomography. AB - BACKGROUND: The diagnostic accuracy of technetium-99m methoxyisobutylisonitrile (Tc-MIBI) single photon emission computed tomography (SPECT) and computed tomography (CT) of the head and neck for differentiating recurrent or residual nasopharyngeal carcinomas (NPC) from benign lesions after radiotherapy was compared with positron emission tomography (PET) with 18-fluoro-2-deoxyglucose (FDG). METHODS: Thirty six NPC patients underwent head and neck CT, Tc-MIBI SPECT, and FDG-PET four months after radiotherapy to differentiate recurrent or residual NPC from benign lesions. Histopathologic examinations of nasopharyngeal biopsies were performed for all 36 patients. No patients had multiple foci of NPC. RESULTS: Based on biopsy results, the sensitivity, specificity, and accuracy of CT for differentiating recurrent or residual NPC from benign lesions were 73%, 88%, and 83%, respectively. The sensitivity, specificity, and accuracy of Tc-MIBI SPECT were 73%, 96%, and 89%, respectively. The sensitivity, specificity, and accuracy of FDG-PET were 100%, 96%, and 97%, respectively. Combination CT and Tc MIBI SPECT for 28 NPC patients with congruent results showed the same sensitivity, specificity, and accuracy of 100%, 96%, and 96%, respectively, as FDG-PET for differentiating recurrent or residual NPC from benign lesions. In eight patients with incongruent results between CT and Tc-MIBI SPECT, FDG-PET correctly differentiated two benign lesions and six recurrent or residual NPCs. CONCLUSIONS: In detecting recurrent or residual NPC, FDG-PET is the best tool. However, combined use of CT and Tc-MIBI SPECT can result in the same accuracy as FDG-PET. PMID- 11932901 TI - Definition of three minimal deleted regions by comprehensive allelotyping and mutational screening of FHIT,p16(INK4A), and p19(ARF) genes in nasopharyngeal carcinoma. AB - BACKGROUND: Recurrent deletion on a chromosomal location in tumor cells can be detected by frequent allelic loss and generally is considered to be an indication of the existence of a tumor suppressor gene (TSG) in the region. In the current study, using fluorescent-labeled, high-density microsatellite markers for allelotyping, the authors pinpointed three minimal deleted regions (MDRs) and screened mutations of putative TSGs on chromosomes 3, 9, and 11 in nasopharyngeal carcinoma (NPC) cases occurring in Taiwan. METHODS: A total of 133 informative microsatellite markers were used on chromosomes 3, 9, and 11 with an average marker density of 4 centimorgans (cM) for the allelotyping of genomic DNAs isolated from NPC tissues and their corresponding lymphocytes in 48 patients. The correlation between allelic loss and the clinicopathologic parameters of NPC tissues was examined. In addition, putative TSGs including FHIT, p16(INK4a), and p19(ARF) were selected for mutation screening to investigate their potential participation in NPC tumorigenesis. RESULTS: Of 3787 informative allelotyping data, 25 frequent allelic losses (or loss of heterozygosity [LOH]) in 13 cytogenetic loci were identified based on a deletion frequency that was greater than the average of allelic loss on that particular chromosome. Several significant associations were determined after statistical analysis of the correlation between allelic loss and clinicopathologic parameters. The allelic losses by D9S318 and D11S1304 were associated with N2/N3 (P = 0.035 and P = 0.005, respectively), and those by D9S905 and D11S1304 were associated with grouped American Joint Committee on Cancer (AJCC) Stage III/IV samples (P = 0.022 and P = 0.017, respectively) of NPC tissues. In addition, three MDRs were revealed on 3p25.3-24.1 (< 19 cM), 3p23-21.31 (< 9 cM), and 11q22.1-23.2 (< 8 cM). To examine somatic mutations in previously reported TSGs located near these frequent LOH loci, three candidate genes, p16(INK4a), p19(ARF), and FHIT, were analyzed. Point mutations in the coding region of FHIT and in the intron 1 splicing acceptor site of both p16(INK4a) and p19(ARF) were detected in NPC cell lines. Sequence analysis of both p16(INK4a) and p19(ARF) transcripts revealed that the point mutation resulted in skipping of exon 2 and the generation of shorter transcripts. CONCLUSIONS: High-density allelotyping permitted the discovery of 3 MDRs on 3p25.3-24.1 (< 19 cM), 3p23-21.31 (< 9 cM), and 11q22.1 23.2 (< 8 cM) and a correlation was determined between allelic loss and clinicopathologic parameters of NPC tissues. More important, one somatic mutation in NPC cell lines on the intron 1/exon 2 splicing acceptor site of the INK4a/ARF locus was found to result in exon 2 skipping both p16(INK4a) and p19(ARF) transcripts, which presumably inactivates the functions of both the p16(INK4a) and p19(ARF) proteins. PMID- 11932902 TI - Molecular pathology parameters in human nasopharyngeal carcinoma. AB - BACKGROUND: To derive a better understanding of the biologic behavior of nasopharyngeal carcinoma (NPC), the authors evaluated a number of molecular variables to address the hypothesis that p53 dysfunction in NPC is associated with Epstein-Barr virus (EBV), increased tumor angiogenesis, lower likelihood of apoptosis, and poorer clinical outcome. MATERIALS: The biopsy samples from 87 NPC patients were obtained and sections were made to detect EBV, using in-situ hybridization; the authors used immunohistochemistry to assess p53, p21(WAF1/CIP1) expression, and microvessel density count (MVD). In situ end labelling was used to evaluate apoptosis and necrosis. Analyses were conducted on the association between each of these variables as well as clinical outcome, including survival and local control. RESULTS: There was a highly significant association between EBV-encoded RNA (EBER) positivity with p53 over-expression in that only 1 out of 32 p53 over-expressing tumors was EBER negative, as opposed to 19 out of 48 p53 negative tumors being EBER negative (P = 0.001). In addition, EBER positivity was highly associated with World Health Organization (WHO) type 3 NPC, Asian/Chinese ethnicity, a lower apoptotic index, and p21 over-expression. p53 over-expression was associated with a higher MVD count. Controlling for age and nodal status, EBER positivity was associated with both improved overall survival (P = 0.02), and disease-free survival (P = 0.04). In contrast, the presence of tumor necrosis was associated with an inferior local control (P = 0.03). CONCLUSION: p53 protein was over-expressed in approximately one third of NPC samples in the current study, and this correlated significantly with the presence of EBER. Epstein-Barr virus status was also associated with WHO type 3 NPC, Asian/Chinese ethnicity, and induction of p21. The presence of EBV appeared to predict for improved survival, the mechanism of which remains to be elucidated in this biologically complex disease. PMID- 11932903 TI - Site specific dependency of second primary cancer in early stage head and neck squamous cell carcinoma. AB - BACKGROUND: A study was conducted to determine the site specificity of second primary cancer (SPC) following early (Stage I and II) squamous cell carcinoma of the head and neck (SCH) and its impact on prognosis. METHODS: Between 1956 and 1999, 1,639 early stage SCHs in 1,609 patients were treated by radiotherapy at the authors' institution. The risk of additional respiratory and upper digestive tract (RUDT) carcinoma and the treatment outcome were reviewed for each SCH site. Second primary RUDT carcinomas were the second most common cause of death (113 cases) after death from primary SCH carcinomas (350 cases). RESULTS: Among the 1,609 early stage SCH patients, 333 second primary cancers were documented in 258 patients, 235 (71%) of which were RUDT carcinomas. The risk of SPCs was 3.2% per year, and the risk of SPCs in the RUDT region was 2.3% per year. The risk of SPCs was almost two times higher in males (3.7% per year) than in females (2.2% per year). The highest risk of SPC was observed in the oro-hypopharynx index cancer group (8.5% per year), and the lowest risks were observed in the epipharynx group (0% per year) and the maxillary sinus group (1.4% per year). CONCLUSIONS: Different SPC risks according to the SCH site result in a different prognosis for each site of early stage SCH. Long-term follow-up care after the initial treatment for early SCH should be continued according to the site of the initial SCH. PMID- 11932904 TI - Changing incidence of non-Hodgkin lymphomas in the United States. AB - BACKGROUND: The incidence of non-Hodgkin lymphoma (NHL) has been rising in many regions and populations during the last few decades. Data from the Surveillance, Epidemiology, and End Results (SEER) Program show that age-adjusted rates of NHL increased through the 1980s but leveled off in the 1990s. METHODS: To determine whether the incidence of NHL stabilized in all population subgroups, particularly in age-defined groups with distinctive risks of NHL, the authors investigated trends in NHL incidence among persons aged 0-14 years, 15-24 years, 25-34 years, 35-44 years, 45-54 years, 55-64 years, 65-74 years, and > 75 years by gender and race using 1973-1998 data from the SEER Program, which covered approximately 10% of the U.S. population. Joinpoint regression was used to assess changes in trends across the period. RESULTS: NHL incidence trends changed significantly among males aged 25-54 years, in whom rates began to decrease (6-16% per year) in the middle to late 1990s, as well as among most whites aged > or = 55 years, in whom rate increases slowed from 3-4% to 1-2% per year in the late 1980s. Incidence trends were steady in other groups, with uniform increases among whites aged 15 24 years (2-3% per year), women aged 25-54 years (1-6% per year), and blacks aged > or = 55 years (2-4% per year). Although recent age specific incidence rates were generally higher in males compared with females and in whites compared with blacks, among males aged 25-54 years, rates were significantly higher in black males compared with white males. CONCLUSIONS: There have been changes in the demographic groups impacted by NHL. The trends for human immunodeficiency virus probably are related to recent decreases in NHL incidence among males aged 25-54 years. The rate change in the older white population is unexplained but represents both an alleviation of the burden of NHL in this population and a potential opportunity to generate hypotheses regarding risk factors for the development of NHL. PMID- 11932905 TI - Simultaneous homoharringtonine and interferon-alpha in the treatment of patients with chronic-phase chronic myelogenous leukemia. AB - BACKGROUND: Homoharringtonine (HHT) has antileukemic activity in patients with Philadelphia chromosome (Ph) positive chronic myelogenous leukemia (CML). Combinations of HHT, interferon-alpha (IFN-alpha), and cytarabine (ara-C) have been studied in various CML phases. The objectives of this study were to evaluate the efficacy and toxicity profiles of a combination regimen of simultaneous HHT and IFN-alpha therapy in patients with chronic-phase CML who were not exposed previously to either agent. METHODS: Forty-seven patients were treated: 37 patients with early chronic-phase CML (2 patients with clonal evolution) and 10 patients with late chronic-phase CML. Their median age was 62 years (range, 23-73 years). HHT was given at a dose of 2.5 mg/m(2) by continuous intravenous infusion over 24 hours daily for 5 days every month, and IFN-alpha was given daily at a target dose of 5 x 10(6) units/m(2) subcutaneously. Response, survival, and treatment toxicity were analyzed. RESULTS: Overall, the complete hematologic response (CHR) rate was 85%; the cytogenetic response rate was 66%, with major cytogenetic responses (Ph positive in < 35% of metaphases) in 49% of patients and complete cytogenetic responses in 21% of patients. The CHR rate, cytogenetic response rate, and major cytogenetic response rate were 84%, 69%, and 52%, respectively, in patients with early chronic-phase CML. Among the 10 patients with late chronic-phase CML, the CHR rate, cytogenetic response rate, and major cytogenetic response rate were 80%, 50%, and 40%, respectively. Response rates in patients age > 60 years were 84%, 62%, and 49% for CHR, cytogenetic response, and major cytogenetic response. Myelosuppression was frequent but manageable: Anemia with hemoglobin < 8.0 g/dL occurred in 36% of patients, requiring dose adjustments and erythropoietin therapy. Nonhematologic toxicities were mainly fatigue, aches, and gastrointestinal disturbances. Dose reductions with multiple courses were significant and were due to myelosuppression: After 6-24 courses, the median daily IFN-alpha dose was 1 MU/m(2), and the median number of days on HHT per month was 2 days. With a median follow-up of 26 months, the estimated 2 year survival rate was 90% (95% confidence interval, 79-100%). CONCLUSIONS: The simultaneous combination of HHT and IFN-alpha is safe and effective, but the dose schedules that actually were delivered were significantly lower than the planned dose schedules. With the availability of signal-transduction inhibitor 571 (imatinib mesylate), studies of combination of HHT and IFN-alpha chemotherapy in patients with CML who have disease that fails to respond to imatinib mesylate and of combinations with imatinib mesylate need to be explored. PMID- 11932906 TI - Frequency and type of serious infections in fludarabine-refractory B-cell chronic lymphocytic leukemia and small lymphocytic lymphoma: implications for clinical trials in this patient population. AB - BACKGROUND: Treatments for fludarabine-refractory chronic lymphocytic leukemia (CLL)/small lymphocytic lymphoma (SLL) are limited. Most new therapies being examined in fludarabine-refractory patients have shown a high frequency of serious infection. Little data exist regarding the frequency of infections in this population treated with noninvestigational best supportive care therapies. METHODS: The infectious courses of 27 patients with fludarabine-refractory CLL/SLL were retrospectively reviewed. Fludarabine-refractoriness was defined as either relapse within six months of completion of or failure to respond to fludarabine treatment. Infections were documented after patients met National Cancer Institute criteria for further treatment. Serious infections were defined as infections mandating admission to the hospital for intravenous antibiotics. RESULTS: Patient characteristics included: median age 67 years (range, 40-83), median 3 chemotherapy treatments (range, 1-8), and hypogammaglobulinemia in 73% of patients. Pneumocystis carinii prophylaxis was given to 89% of patients. Serious infections developed in 24 out of 27 patients (89%). Patients had a median of 2 admissions (range, 0-11) for serious infection occurring at a median of 4 months (range, 0-21) from onset of fludarabine-refractoriness. The median frequency of admission for infection was 0.17 per month. Most common sites for infection in decreasing frequency were: respiratory tract, urinary tract, blood/sepsis, and soft tissues. Bacteria caused 69 out of 88 infections (78.4%); viruses (varicella-zoster and herpes simplex) caused 11 out of 88 (12.5%); fungi caused 4 out of 88 (4.5%); and opportunistic infections caused 4 out of 88 (4.5%). Median survival was 13.0 months (range, 1-44+). CONCLUSIONS: The frequency of serious infections in patients with fludarabine-refractory CLL/SLL is high. These findings are relevant to trials with new and highly effective agents for which the incidence of serious infections after treatment might otherwise appear to be prohibitively high. PMID- 11932907 TI - Combined hepatocellular and cholangiocarcinoma: demographic, clinical, and prognostic factors. AB - BACKGROUND: Tumors with combined hepatocellular and cholangiocellular features are well known histopathologically but their clinical behavior is poorly understood. The objectives of the current study were to define the demographic profile of the patients in whom these uncommon tumors occur and to evaluate treatment outcome in comparison with that in patients with either hepatocellular carcinoma (HCC) or peripheral cholangiocarcinoma (CC) alone. METHODS: Twenty seven patients with combined tumors were identified from a prospective database. Pathologic specimens were analyzed to confirm the diagnosis. Demographics, clinical data, and survival were analyzed. Outcome after resection was compared with that of patients with CC and with a matched group of patients with HCC. RESULTS: The gender distribution of the combined tumors (52% men and 48% women) was intermediate between HCC (67% men and 33% women) and CC (30% men and 70% women) (P = 0.03). The incidence of positive hepatitis B or C serology and cirrhosis was similar in patients with combined tumors and those with CC (15% and 0% vs. 13% and 4%, respectively); similarly, patients of Asian heritage constituted 7% and 9%, respectively, of the patients with these tumors. By contrast, cirrhosis (41%) and positive hepatitis serology (56%) were far more common in patients with HCC, and 19% of these patients were of Asian heritage. Twenty-one of 27 patients with combined tumors (78%) underwent resection. All 6 patients with combined tumors that were not amenable to resection died of disease within 18 months. After resection, the 5-year survival was lowest in patients with combined tumors (24%) but was not significantly different from that in patients with CC (33%) or HCC (37%). The liver was the most common site of recurrence in all three groups. CONCLUSIONS: The demographic and clinical features of patients with combined tumors were most similar to those of patients with CC. Most important, combined tumors were not found to be associated with chronic liver disease; consequently, the resectability rate was higher for these tumors than typically is reported for HCC. Resection was associated with long term survival in some patients, but recurrent hepatic disease was common. The presence of cholangiocellular differentiation appeared to worsen the prognosis when compared with pure HCC, although this difference did not reach statistical significance. PMID- 11932909 TI - Trimodality treatment in Stage III nonsmall cell lung carcinoma: prrognostic impact of K-ras mutations after neoadjuvant therapy. AB - BACKGROUND: In a trimodality treatment approach for Stage III nonsmall cell lung carcinoma (NSCLC), the prognostic impact of the ras mutation status in resection specimens was evaluated. METHODS: Forty patients with Stage III NSCLC underwent tumor resection after neoadjuvant treatment with two cycles of chemotherapy (ifosfamide, carboplatin, and etoposide) and subsequent twice-daily radiotherapy (45 grays [Gy]; 2 x 1.5 Gy/day) with concurrent carboplatin and vindesine. Assessment of K-ras codon 12 mutation status was performed in the paraffin embedded resection specimens by a two-step polymerase chain reaction followed by restriction fragment length polymorphism analysis. RESULTS: K-ras mutation status could be assessed in 28 cases. A K-ras codon 12 point mutation was found in 13 of 28 resection specimens (46%). The mutation was found independently of gender, age, tumor stage, and clinical response status and occurred more frequently in adenocarcinomas. Even after complete resection, the presence of a K-ras mutation was a significant predictor for a poor progression free survival (P = 0.005). CONCLUSIONS: These data suggest that further evaluation of the K-ras codon 12 mutation status in trials on neoadjuvant and adjuvant therapy is warranted. This may contribute to the identification of stratification variables for future treatment approaches. PMID- 11932908 TI - Frequent impairment of the spindle assembly checkpoint in hepatocellular carcinoma. AB - BACKGROUND: Chromosomal instability (CI) leading to aneuploidy is one form of genetic instability, a characteristic feature of various types of cancers. Recent work has suggested that CI can be induced by a spindle assembly checkpoint defect. The aim of the current study was to determine the frequency of a defect of the checkpoint in hepatocellular carcinoma (HCC) and to establish whether alterations of genes encoding the checkpoint were associated with CI in HCC. METHODS: Aneuploidy and the function of the spindle assembly checkpoint were examined using DNA flow cytometry and morphologic analysis with microtubule disrupting drugs. To explore the molecular basis, the authors examined the expression and alterations of the mitotic checkpoint gene, BUB1, using Northern hybridization and direct sequencing in 8 HCC cell lines and 50 HCC specimens. Furthermore, the authors examined the alterations of other mitotic checkpoint genes, BUBR1, BUB3, MAD2B, and CDC20, using direct sequencing in HCC cell lines with aneuploidy. RESULTS: An impaired spindle assembly checkpoint was found in five (62.5%) of the eight aneuploid cell lines. Transcriptional expressions of the BUB1 gene appeared in all cell lines. While some polymorphic base changes were noted in BUB1, BUBR1, and CDC20, no mutations responsible for impairment of the mitotic checkpoint were found in either the HCC cell lines or HCC specimens, which suggests that these genes did not seem to be involved in tumor development in HCC. CONCLUSIONS: The loss of spindle assembly checkpoint occurred with a high frequency in HCC with CI. However, other mechanisms might also contribute to CI in HCC. PMID- 11932910 TI - Prediction of intracranial metastases in cancer patients with headache. AB - BACKGROUND: The current study was conducted to investigate the diagnostic value of neurologic evaluation for the prediction of intracranial metastases in cancer patients with new or changed headache. METHODS: Between February 1997 and February 2000, general practitioners and specialists referred cancer patients with new or changed headache to the Department of Neurology at the study institution. All patients underwent a structured history and neurologic examination. Magnetic resonance imaging (MRI) of the brain was used as the gold standard for determining the presence of intracranial metastases. The association between baseline patient characteristics, history variables, and variables from the neurologic examination in patients with intracranial metastases was evaluated by univariate and multivariate logistic regression analyses in combination with receiver operating characteristic (ROC) curve analyses. RESULTS: Sixty-eight consecutive patients were included in the current study (48 females and 20 males). The mean age of the patients was 57 years (range, 24-88 years; standard deviation +/- 13.3 years). Breast carcinoma was the primary tumor in 32 patients (47.1%) and lung carcinoma was the primary tumor in 12 patients (17.6%). Intracranial metastases occurred in 22 patients (32.4%). The occurrence of intracranial metastases was predicted in the multivariate logistic regression analyses by one baseline patient characteristic variable and 2 history variables (i.e., headache duration of < or =10 weeks [odds ratio (OR) of 11.0; 95% confidence interval (95% CI), 1.1-108.2], emesis [OR of 4.0; 95% CI, 1.1-14.3], and pain not of tension- type [OR of 6.7; 95% CI, 1.8-25.1]). No variable from the neurologic examination was found to add information to the prediction model. When at least one of the three predictors was present, all patients with intracranial metastases could be identified with this prediction model. MRI could be omitted in 12 patients (26%) without intracranial metastases. The ROC area under curve of this model was 0.83. CONCLUSIONS: Intracranial metastases were found in 32.4% of the cancer patients with headache as the presenting symptom. Although 3 significant clinical predictors were found (headache duration < or =10 weeks, emesis, and pain not of tension- type), few patients could be excluded from undergoing MRI because of a low specificity. Therefore, MRI of the brain was considered to be warranted in all patients in the current study. PMID- 11932912 TI - Treatment decisions for breast carcinoma: patient preferences and physician perceptions. AB - BACKGROUND: Patient autonomy and participation in treatment decision making have been encouraged in recent years. However, patients and physicians frequently disagree with regard to the patient's needs and perceptions of their illness. To the authors' knowledge to date only limited research has assessed physicians' perceptions of patients' decision-making preferences. The purpose of the current prospective study was to determine the agreement between patient decision-making preferences and physician perceptions of those preferences. METHODS: Women with breast carcinoma who were attending their first outpatient consultation with a breast medical oncologist in a university cancer center were enrolled in the current study. At the end of the consultation, the patients were given a survey regarding their treatment decision-making preferences that included active, shared, and passive roles in decision-making and the patients' attending physicians also were given a survey regarding their perceptions of the patients' decision-making preferences. RESULTS: Fifty-seven patients had complete data and were analyzed. Approximately 89% of these 57 patients preferred either an active or a shared role in decision making. The agreement between patients and physicians with regard to decision-making preference only occurred in 24 cases (42%). The majority of covariates such as age, education, and income were not found to be statistically significant with regard to patient preferences or to the proportion of patients and physicians who agreed on the patient's preferences. CONCLUSIONS: Women with breast carcinoma appear to have a strong desire for involvement in making decisions regarding their treatment. However, physicians do not appear to be consistently able to predict the decision-making preferences of their patients. Enhanced agreement between patient preferences and physician expectations mostly likely will improve communication and patient satisfaction with the treatment decision-making process. PMID- 11932911 TI - Tumor specific gene expression profiles in human leiomyosarcoma: an evaluation of intratumor heterogeneity. AB - Leiomyosarcomas are malignant smooth muscle tumors characterized by a spectrum of histopathologic features and clinical behavior. Gene expression profiling of leiomyosarcomas may identify differential gene signatures that may allow for the clinical stratification of the tumors. Typically, surgical specimens from these tumors are large and manifest a variegated macroscopic appearance. Because of their large size at the time of diagnosis, sufficient tissue is available for regional and clonal heterogeneity assessment. However, if the gene expression profiles of samples taken from different locations in the tumors are drastically different, biologic classification on the basis of random sample analysis may not be adequate. Therefore, to assess intertumor and intratumor heterogeneity, the authors performed a gene expression study using leiomyosarcoma specimens from three excised sarcomas from an equal number of different patients. Comparisons between tumor and normal samples from the three patients as well as between carefully mapped peripheral and core specimens from the same tumor (excised from one of the patients), were performed. Analysis of the expression profiles demonstrated minimal intratumor variations compared with intertissue variations, indicating homogeneous tumor specific gene expression profiles. The authors also identified genes that are expressed differentially in tumor and normal tissue. PMID- 11932913 TI - High incidence of thyroid dysfunction despite prophylaxis with potassium iodide during (131)I-meta-iodobenzylguanidine treatment in children with neuroblastoma. AB - BACKGROUND: Treatment modalities like targeted radiotherapy with (131)I-meta iodobenzylguanidine ((131)I-MIBG) improve survival rates after neuroblastoma (NB). Radiation to the thyroid gland can lead to hypothyroidism and even malignancy. Because hypothyroidism after (131)I-MIBG treatment was reported, the current KI prophylaxis against thyroidal radiation damage was evaluated. METHODS: The incidence, pathogenesis, and consequences of thyroid dysfunction among 42 NB patients treated with (131)I-MIBG were evaluated retrospectively. Efficacy of KI prophylaxis was established by measuring thyroidal radioiodide uptake. Thyroid damage was expressed as thyrotropin elevation (TE, plasma concentration of thyroid stimulating hormone > or = 4.5 mU/L). RESULTS: The mean followup was 2.3 years (range, 0.1-8.5). The mean number of treatments with (131)I-MIBG was 3.3. Of 428 scintigrams, uptake of (131)I in the thyroid was visible in 92 (21.0%). Twenty two patients (52.4 %) presented TE after a mean period of 1.4 years (range, 0.1-5.8). Clinical signs of hypothyroidism were not observed. Eight patients received suppletion therapy with thyroxine. Thyrotropin elevation was transient in four patients. Of 25 survivors, with a mean followup of 3.5 years, 16 (64%) developed TE. No correlation was found between TE and thyroid visualization after (131)I-MIBG administration or the number of treatments. No abnormalities were seen by ultrasound imaging of the thyroid. CONCLUSIONS: Occurrence of thyroid dysfunction after treatment with (131)I-MIBG for NB is high, in spite of KI prophylaxis. Close followup of thyroid function and structure is required in patients treated with (131)I-MIBG. New ways of protecting the thyroid during exposure to radioiodine should be developed. PMID- 11932914 TI - The PedsQL in pediatric cancer: reliability and validity of the Pediatric Quality of Life Inventory Generic Core Scales, Multidimensional Fatigue Scale, and Cancer Module. AB - BACKGROUND: The Pediatric Quality of Life Inventory (PedsQL) is a modular instrument designed to measure health-related quality of life (HRQOL) in children and adolescents ages 2-18 years. The PedsQL 4.0 Generic Core Scales are multidimensional child self-report and parent proxy-report scales developed as the generic core measure to be integrated with the PedsQL disease specific modules. The PedsQL Multidimensional Fatigue Scale was designed to measure fatigue in pediatric patients. The PedsQL 3.0 Cancer Module was designed to measure pediatric cancer specific HRQOL. METHODS: The PedsQL Generic Core Scales, Multidimensional Fatigue Scale, and Cancer Module were administered to 339 families (220 child self-reports; 337 parent proxy-reports). RESULTS: Internal consistency reliability for the PedsQL Generic Core Total Scale Score (alpha = 0.88 child, 0.93 parent report), Multidimensional Fatigue Total Scale Score (alpha = 0.89 child, 0.92 parent report) and most Cancer Module Scales (average alpha = 0.72 child, 0.87 parent report) demonstrated reliability acceptable for group comparisons. Validity was demonstrated using the known-groups method. The PedsQL distinguished between healthy children and children with cancer as a group, and among children on-treatment versus off-treatment. The validity of the PedsQL Multidimensional Fatigue Scale was further demonstrated through hypothesized intercorrelations with dimensions of generic and cancer specific HRQOL. CONCLUSIONS: The results demonstrate the reliability and validity of the PedsQL Generic Core Scales, Multidimensional Fatigue Scale, and Cancer Module in pediatric cancer. The PedsQL may be utilized as an outcome measure in clinical trials, research, and clinical practice. PMID- 11932915 TI - Increased proliferative activity caused by loss of p21(WAF1/CIP1) expression and its clinical significance in patients with early-stage gastric carcinoma. AB - BACKGROUND: Recurrences of gastric carcinoma are likely to take on a variety forms, even after patients undergo curative resection for early-stage gastric carcinoma. It is important to identify the biologic markers that predict tumor progression and survival in these patients. Proliferating cell nuclear antigen (PCNA) acts as a processivity factor for DNA polymerase delta, which is involved directly in DNA synthesis, and the PCNA level is correlated with the proliferative state of cells. p21(WAF1/CIP1) interacts with PCNA to inhibit DNA synthesis and plays a central role in regulating the cell cycle. The authors investigated patients with early-stage gastric carcinoma to determine the clinical significance of proliferative activity and p21 expression. METHODS: Tissue specimens from 133 Japanese patients with early-stage gastric carcinoma that invaded the submucosal layer were immunostained with a monoclonal antibody against PCNA and p21(WAF1/CIP1), and the correlations between the PCNA labeling index and p21(WAF1/CIP1) expression as well as clinicopathologic factors were investigated. RESULTS: The PCNA labeling index varied from 9.9% to 81.4%, (mean, 31.2%). The incidence of p21 positive expression was 87 of 133 patients (65.4%). The patients with a high labeling index had a significantly higher rate of lymph node metastasis (P < 0.01) and loss of p21(WAF1/CIP1) expression (P < 0.05) compared with the patients with a low labeling index. The 5-year survival rate for patients in the high labeling index group (87.0%) was significantly lower compared with the 5-year survival rate for the patients in the low labeling index group (98.6%; P < 0.05). CONCLUSIONS: Loss of p21(WAF1/CIP1) expression contributes to the amplification of proliferative activity in patients with early stage gastric carcinoma. Estimation of the proliferative activity of early-stage gastric carcinoma provides information on lymph node metastasis and prognosis. Even after patients undergo curative resection, those with early-stage gastric carcinoma should be followed closely. PMID- 11932916 TI - National action plan on Breast Cancer Workshop on Multicultural Aspects of Breast Cancer Etiology: recommendations of the workshop. PMID- 11932917 TI - Clinicopathologic implications of E-cadherin reactivity in patients with lobular carcinoma in situ of the breast. PMID- 11932920 TI - Induction of cell death in T lymphocytes by invasin via beta1-integrin. AB - Ligand binding to beta1-integrins exerts multiple effects on cells of the immune system including adhesion, spreading, haptotaxis and costimulation of T cells activated by anti-CD3. Here we show that a high-affinity ligand for beta1 integrins, the invasin (Inv) protein of Yersinia pseudotuberculosis, can induce cell death in T lymphocytes via a rapid process. Partially purified native Inv protein and an Inv fusion protein caused apoptotic/necrotic caspase-independent cell death in T lymphocytes as determined by phosphatidylserine exposure on the cell surface, uptake of propidium iodide, labeling of DNA strand breaks and presence of DNA ladder. Inv-induced cell death was mediated via beta1-integrins as indicated by the fact that Inv bound to the beta1-integrin subunit (CD29), that anti-beta(1)-integrin antibodies blocked Inv-induced cell death and that Inv induced cell death was absent in two beta1-integrin- cell lines produced by different procedures. Killing via beta1-integrins represents a novel pathway for cell death in T lymphocytes. PMID- 11932921 TI - The M603 idiotype is lost in the response to phosphocholine in terminal deoxynucleotidyl transferase-deficient mice. AB - The majority of anti-phosphocholine (PC) antibodies induced by the PC epitope in Proteus morganii (PM) express the M603 idiotype (id), which is characterized by an invariant Asp to Asn substitution at the V(H):D(H) junction. To elucidate the molecular basis by which M603-like B cells acquire the mutations resulting in this invariant substitution, we analyzed the immune response to PC-PM in terminal deoxynucleotidyl transferase (TdT) gene knockout (KO) mice. In the absence of TdT, T15-id antibodies comprised 80-100% of the primary response to PC-PM. Less than 10% of the response in wild-type mice is T15-id(+). In TdT KO mice, the secondary response to PC-KLH was higher than in wild-type mice and was dominated by the germ-line T15-id. About 10% of this response, in both TdT KO and wild-type mice, comprised M167-id(+) antibodies. Additionally, none of the functionally rearranged V1/DFL16.1/J(H)1 cDNA isolated from PC-PM-immunized TdT KO mice showed the Asp/Asn substitution characteristic of PC-binding, PC-PM-induced M603-like antibodies. These data indicate that production of M603-id antibody is TdT dependent, while generation of M167-id antibody is TdT independent, and that in the absence of competition from M603-like B cells, T15-id B cells can respond to PC-PM. PMID- 11932922 TI - Transgenic B lymphocytes expressing a human cold agglutinin escape tolerance following experimental infection of mice by Mycoplasma pulmonis. AB - Several microbial infections, including Mycoplasma pneumoniae respiratory infection, are capable, in man, of transiently inducing the expression of anti red blood cell autoantibody called cold agglutinins (CA). To analyze the mechanisms by which immune tolerance is broken following a mycoplasma infection, we used transgenic mice expressing a pathogenic human CA, designated CA-GAS, specific for sialylated carbohydrates. In these mice peripheral deletion of autoreactive B lymphocytes and receptor editing, prevent the development of autoimmune hemolytic anemia. Experimental infections of transgenic mice with Mycoplasma pulmonis resulted in a high anti-mycoplasma antibody response (despite a severe B cell depletion at the onset of infection), and an important induction of serum CA concentrations, reaching in some mice pathological titers. Whereas in naive mice, only a small percentage of CA-expressing cells could be detected, in infected mice, a majority of circulating B lymphocytes were large B220(-) cells, which expressed the transgenic immunoglobulin. Immunization of the transgenic mice with keyhole limpet hemocyanin and Freund's adjuvant, to nonspecifically stimulate the expression of the passenger transgenes, only moderately increased the CA titers. These results indicate that M. pulmonis infection is capable of breaking immune tolerance in the CA-transgenic mice, in part through specific activation of CA-expressing B lymphocytes. This experimental infection mimics the induction of CA in humans and provide an animal model for studying the genesis of the autoimmune hemolytic anemia. PMID- 11932923 TI - Disruption of the C5a receptor gene fails to protect against experimental allergic encephalomyelitis. AB - Activation of the complement system generates the anaphylatoxic peptide C5a, which elicits a broad range of inflammatory activities. The biological activities of C5a are mediated through its binding to the widely expressed C5a receptor (C5aR), a G-protein-coupled seven transmembrane domain receptor. In experimental autoimmune encephalomyelitis (EAE), an animal model for multiple sclerosis, the C5aR is expressed on monocytes/macrophages, reactive astrocytes and T cells infiltrating the central nervous system (CNS). To investigate the role of the C5aR in this T cell-driven autoimmune model, we induced EAE in C5aR-deficient mice (C5aR(-/-)) and wild-type mice using a myelin oligodendrocyte glycoprotein (MOG) peptide as the immunogen. We found that C5aR(-/-) mice were fully susceptible to MOG-induced EAE with no difference in disease onset or severity in C5aR(-/-) mice compared to control mice. Cellular infiltrates (macrophages and T cells) were similar in the spinal cords of both animal groups and splenic T cells from C5aR(-/-) mice and control mice responded identically to MOG in T cell proliferation assays. Ribonuclease protection assays demonstrated no significant differences in pro-inflammatory gene expression between receptor-deficient and sufficient mice. These results indicate that the C5aR is not an essential mediator in the induction and progression of EAE. PMID- 11932924 TI - The efficiency of B cell receptor (BCR) editing is dependent on BCR light chain rearrangement status. AB - Anti-DNA knock-in mice serve as models for studying B cell tolerance mechanisms to a ubiquitous antigen. We have constructed six strains of double transgenic (C57BL/6xBALB/c)F1 mice, each expressing an unmutated or somatically mutated anti DNA heavy (H) chain, combined with one of three different light (L) chains, namely V(kappa)1-J(kappa)1, V(kappa)4-J(kappa)4 and V(kappa)8-J(kappa)5. In vitro analysis of the various Ig H/L chain combinations showed that all had a similar specificity for single-stranded DNA and double-stranded DNA, but that antibodies encoded by the mutated H chain had higher affinities for the autoantigen. None of the targeted mouse strains exhibited significant levels of serum anti-DNA activity. However, while B cells from mice carrying the V(kappa)1-J(kappa)1 transgenic L chains were tolerized almost exclusively by L chain receptor editing in an affinity-independent manner, the mice expressing V(kappa)8-J(kappa)5 L chains have utilized affinity-dependent clonal anergy as their sole mechanism of B cell tolerance. V(kappa)4-J(kappa)4 targeted mice exhibited an intermediate phenotype with respect to these two mechanisms of B cell tolerance. Our results suggest that receptor editing is the preferred mechanism of B cell tolerance and that the efficiency of L chain editing is directly related to the number of available J(kappa) segments on the expressed V(kappa) allele. PMID- 11932925 TI - Thymopoiesis requires Pax9 function in thymic epithelial cells. AB - The epithelial thymic anlage develops from the third pharyngeal pouch. Pax9 is expressed in the entire pharyngeal endoderm, and its function is required for normal development of organs derived from pharyngeal pouches. Here, we show that in Pax9 null mice, the thymic anlage develops as an ectopic polyp-like structure in the larynx. It expresses Whn/Foxn1, a marker of thymic epithelium, but fails to perform the normal caudo-ventral movement to the upper mediastinum. The thymic rudiment contains mesenchymal cells, blood vessels and is colonized by T cell progenitors. However, from embryonic day 14.5 onwards, the size of the Pax9 mutant thymus is severely reduced. Whereas expression of TCRbeta chain genes is readily detectable in the mutant thymus, no expression of the TCRgamma chain was detectable. Our results identify a new genetically defined control point of thymopoiesis. PMID- 11932926 TI - Toll-like receptor 1 inhibits Toll-like receptor 4 signaling in endothelial cells. AB - Toll-like receptor 4 (TLR4) is the signal-transducing component of the LPS recognition complex and is essential for LPS-induced septic shock. Here we demonstrate that TLR1 has the capacity to abrogate TLR4 signaling. Human microvascular endothelial cells express TLR4 but not TLR1 and respond to LPS through TLR4. The ability of these cells to respond to LPS was lost, however, when they were transfected with TLR1. Inhibition was specific for TLR1 because TL5 failed to block TLR4 function. Moreover, TLR1 had no effect upon TNF-alpha signaling, indicating that TLR1 operated at a step upstream of the convergence between the two pathways. Inhibition of TLR4 signaling was mediated by the extracellular, but not cytoplasmic domain of TLR1. In addition, TLR1 physically associated with TLR4 in co-precipitation experiments. These findings suggest that TLR1 might restrain potentially dangerous innate response to LPS by binding to TLR4 and preventing the formation of active signaling complexes. PMID- 11932927 TI - Non-LPS components of Chlamydia pneumoniae stimulate cytokine production through Toll-like receptor 2-dependent pathways. AB - Recent studies suggest that infection with Chlamydia pneumoniae is associated with atherosclerosis, and that cytokines play an important role in the initiation and progression of Chlamydia-induced inflammation. When freshly isolated peripheral blood mononuclear cells (PBMC) were stimulated for 24 h with sonicated C. pneumoniae, significant amounts of the pro-inflammatory cytokines TNF-alpha and IL-1beta and of the anti-inflammatory cytokine IL-10 were released into the supernatant. The addition of serum increased cytokine release induced by C. pneumonia two- to fivefold (p < 0.01). This effect was not due to complement, mannose-binding lectin (MBL) or lipopolysaccharide-binding protein (LBP). Incubation of PBMC with either anti-Toll-like receptor 4 (TLR4) or anti-CD14 blocking antibodies did not influence the production of cytokines induced by Chlamydia. The induction of cytokines by C. pneumoniae in macrophages from C3H / HeJ mice, known to have a defective TLR4, was identical to that measured in control macrophages from C3H / HeN mice. In contrast, incubation of PBMC with an anti-TLR2 blocking antibody significantly inhibited the production of TNF by 67 % and of IL-1beta by 72 %. In conclusion, C. pneumoniae stimulates cytokine production in a serum-dependent manner, but independently of complement, MBL and LBP. C. pneumoniae induces the pro-inflammatory cytokines TNF and IL-1beta through TLR2, but not TLR4 and CD14. PMID- 11932928 TI - The phosphatidylinositol phosphatase PTEN is under control of costimulation and regulates proliferation in human T cells. AB - The phosphatidylinositol phosphatase gene PTEN is a dual specific phosphatase acting on phospho amino acids but also on three phosphorylated inositol phospholipids. Present results demonstrate that PTEN is inducible by costimulatory signals in human CD4(+) T cells. PTEN expression was up-regulated on RNA and protein level in freshly isolated human CD4(+) T cells following stimulation with CD28 or CD2. In contrast, PTEN expression was high but remained CD28 and CD2 unresponsive in lymphoma cells. Intracellular staining revealed PTEN expression in CD4(+) T cell populations stimulated with anti-CD28 or anti-CD28 / anti-CD3. Stimulation with anti-CD3 alone did not induce PTEN expression. Inhibition of PTEN expression by antisense oligonucleotides in CD4(+) T cells stimulated with non-mitogenic anti-CD28 resulted in massively increased proliferation, which was sensitive to the phosphatidylinositol 3-kinase (PI3 K) inhibitor wortmannin. Although CD28 and CD2 induce PI3 K signal transduction, wortmannin did not block PTEN up-regulation by CD28 or CD2 indicating that PTEN gene expression is PI3 K independent. These results demonstrate that PTEN negatively controls costimulatory signals by antagonizing PI3 K activity in the absence of TCR engagement. PMID- 11932929 TI - Immunohistochemical localization of three different prepro-GnRHs in the brain and pituitary of the European sea bass (Dicentrarchus labrax) using antibodies to the corresponding GnRH-associated peptides. AB - The distribution of the cells expressing three prepro-gonadotrophin-releasing hormones (GnRH), corresponding to salmon GnRH (sGnRH), seabream GnRH (sbGnRH), and chicken GnRH-II (cGnRH-II) forms, was studied in the brain and pituitary of the sea bass (Dicentrarchus labrax) by using immunohistochemistry. To circumvent the cross-reactivity problems of antibodies raised to GnRH decapeptides, we used specific antibodies generated against the different sea bass GnRH-associated peptides (GAP): salmon GAP (sGAP), seabream GAP (sbGAP), and chicken-II GAP (cIIGAP). The salmon GAP immunostaining was mostly detected in terminal nerve neurons but also in ventral telencephalic and preoptic perikarya. Salmon GAP immunoreactive (ir) fibers were observed mainly in the forebrain, although sGAP ir projections were also evident in the optic tectum, mesencephalic tegmentum, and ventral rhombencephalon. The pituitary only receives a few sGAP-ir fibers. The seabream GAP-ir cells were mainly detected in the preoptic area. Nevertheless, sbGAP-ir neurons were also found in olfactory bulbs, ventral telencephalon, and ventrolateral hypothalamus. The sbGAP-ir fibers were only observed in the ventral forebrain, innervating strongly the pituitary gland. Finally, chicken-II GAP immunoreactivity was only detected in large synencephalic cells, which are the origin of a profuse innervation reaching the telencephalon, preoptic area, hypothalamus, thalamus, pretectum, posterior tuberculum, mesencephalic tectum and tegmentum, cerebellum, and rhombencephalon. However, no cIIGAP-ir fibers were detected in the hypophysis. These results corroborate the overlapping of sGAP- and sbGAP-expressing cells in the forebrain of the sea bass, and provide, for the first time, unambiguous information on the distribution of projections of the three different GnRH forms expressed in the brain of a single species. PMID- 11932931 TI - Digital atlases of the antennal lobe in two species of tobacco budworm moths, the Oriental Helicoverpa assulta (male) and the American Heliothis virescens (male and female). AB - The antennal lobe of the moth brain is the primary olfactory center processing information about pheromones and plant odors. We present here a digital atlas of the glomerular antennal lobe structures in the male of Helicoverpa assulta and the male and female of Heliothis virescens, based on synaptic antibody staining combined with confocal microscopy. The numbers of the glomeruli in the three specimens were similar, 65, 66, and 62, respectively. Whereas the male antennal lobe has a macroglomerular complex consisting of three and four units in the two species, the female lobe has two enlarged glomeruli at a corresponding position, near the entrance of the antennal nerve. Another large glomerulus, showing homology in the three specimens, is ventrally located. The small size of the heliothine moths is advantageous for confocal microscopy because the entire brain can be visualized as a single image stack. The maps are freely accessible on the internet, and the digital form of the data allows each atlas to be rotated and sectioned at any angle, providing for the identification of glomeruli in different preparations. PMID- 11932930 TI - Differentiation of tyrosine hydroxylase-synthesizing and/or aromatic L-amino acid decarboxylase-synthesizing neurons in the rat mediobasal hypothalamus: quantitative double-immunofluorescence study. AB - In this double-immunofluorescence study, we first quantified the neurons of the arcuate nucleus as immunoreactive (+) for tyrosine hydroxylase (TH) and/or aromatic L-amino acid decarboxylase (AADC) in rats at embryonic day 21 (E21), at postnatal day 9 (P9), and in adulthood by using conventional fluorescent or confocal microscopy. On E21, monoenzymatic (TH(+)AADC immunonegative (-) and TH( )AADC(+)) neurons and bienzymatic (TH(+)AADC(+)) neurons accounted for 99% and 1%, respectively, of the whole neuron population expressing enzymes of dopamine synthesis. Further development was characterized by the dramatic increase in TH(+)AADC(-) dorsomedial and TH(+)AADC(+) dorsomedial populations from E21 to P9 as well as by the increase in the TH(+)AADC(+) dorsomedial population (in females) and a drop in the TH(+)AADC(-) ventrolateral and TH(+)AADC(-) dorsomedial (in males) populations from P9 to adulthood. In contrast to TH(+)AADC(-) (in males) and TH(+)AADC(+) neurons, the TH(-)AADC(+) neurons did not change in number from E21 to adulthood. Thus, in rat fetuses, the neurons synthesizing TH and/or AADC were mainly monoenzymatic, whereas during postnatal life the fraction of bienzymatic neurons increased by up to 60%. PMID- 11932932 TI - Microglia activation and cell death in response to diethyl-dithiocarbamate acute administration. AB - An increasing body of evidence suggests a role for activated microglia in the pathogenesis of neurodegenerative disorders. Hence, it would be useful to have a better understanding of the significance of microglial activation for neuronal damage. Unfortunately, most models of microglial activation use invasive or long lasting insults, which make it difficult to evaluate the role played by microglia. We have instead developed a model for microglial activation by using brief exposure to the widely available neurotoxin diethyl-dithiocarbamate (DDTC). Despite evidence for the neurotoxic nature of this substance, microglia involvement has not been hitherto investigated. After acute i.p. administration of DDTC at two different doses, microglia were already activated in selected areas of the rat brain (hippocampal dentate gyrus, entorhinal-pyriform cortex and hypothalamus) after 1 hour, reaching a peak at 3-6 hours and subsided within 6-48 hours, depending on the brain region. Microglia activation was associated with interleukin-1 beta immunopositivity between 3 and 6 hours and with up-regulation of major histocompatibility complex class II expression between 24 and 48 hours. No significant changes in astrocyte immunostaining were detected between 6 hours and 6 days. The TUNEL procedure revealed the death of a limited number of cells in the above-mentioned structures that peaked at 6h and then declined rapidly. Cell death was detected in sites with major, minor, or no microglial activation, indicating that these two events can occur concomitantly or independently. The study shows that the administration of DDTC provides a useful model for studying the implications of region-specific reactivity of microglia and its differential interaction with neuronal damage. PMID- 11932933 TI - Hippocampal and prefrontal cortical inputs monosynaptically converge with individual projection neurons of the nucleus accumbens. AB - Afferents to the nucleus accumbens from different sources innervate specific areas of the central "core" and peripheral "shell" and are related to each other, at the light microscopical level, in an intricate overlapping and nonoverlapping way. This lack of homogeneity suggests that this region consists of circuits involving emsembles of neurons modulated by specific sets of convergent afferent inputs and abnormal regulation of such ensembles has been implicated in mental disorders. Early extracellular studies suggested that individual Acb neurons might respond to activation of afferents from more than one excitatory input: More recent studies of hippocampal and amygdalar or prefrontal cortical afferents suggest that hippocampal afferents gate the input from the prefrontal cortex or amygdala. Electrophysiological evidence for convergence of excitatory afferents in the Acb is strong and suggests that these pathways are monosynaptic. Nevertheless, this convergence has proved difficult to demonstrate anatomically as a result of the spatial distribution of the afferent inputs on the dendritic tree of the target neurons. To establish whether individual accumbens neurons receive monosynaptic input from pairs of afferents, one projection was labelled anterogradely with Phaseolus vulgaris leucoagglutinin and the second with biotinylated dextran amine (BDA) with Vector slate grey and 3,3'-diaminobenzidine tetrahydrochloride as the chromagens. Accumbens neurons possibly postsynaptic to these afferents, labelled by an in vivo focal injection of BDA, were examined using correlated light and electron microscopy to establish the proximal-distal distribution of labelled afferent synaptic inputs on their dendritic arbours. Individual cells were shown to receive monosynaptic afferent input from both ventral subiculum and prefrontal cortex, providing an anatomical framework for the hippocampal gating of other limbic inputs to the accumbens. PMID- 11932934 TI - Plastic reaction of the rat visual corticocollicular connection after contralateral retinal deafferentiation at the neonatal or adult stage: axonal growth versus reactive synaptogenesis. AB - The effects of neonatal or adult enucleation on the final adult pattern of the rat visual corticocollicular (C-Co) connection were studied using the anterograde tracer biotinylated dextranamine 10,000 (BDA) iontophoretically injected in the primary visual cortex. In control animals, column-shaped terminal fields limited to a small portion of the collicular surface were observed. Synaptic boutons were present in all superficial strata of the superior colliculus (SC), with the highest density in the ventral part of the stratum griseum superficiale (SGS). Neonatal enucleation caused a considerable expansion of the contralateral visual C-Co terminal fields, which occupied almost the entire collicular surface, suggesting that axonal sprouting had occurred. In addition, terminal boutons tended to localize more dorsally in these cases compared with controls. Following enucleation in adult animals, no changes were observed with respect to the extension of the terminal fields, although a plastic reaction leading to an increase in the bouton density in the stratum zonale (SZ) and upper SGS was found, reflecting a process of reactive synaptogenesis at these levels. These results show that both neonatal and adult visual C-Co fibers react in response to retinal ablation, although this reaction shows distinct characteristics. Molecular factors, such as growth-associated cytoskeletal proteins operating in the cortical origin, and extracellular matrix components and myelin-associated axonal growth inhibitors acting on the collicular target very likely account for these differences. PMID- 11932935 TI - GABA(A) receptor changes in delta subunit-deficient mice: altered expression of alpha4 and gamma2 subunits in the forebrain. AB - The delta subunit is a novel subunit of the pentameric gamma-aminobutyric acid (GABA)(A) receptor that conveys special pharmacological and functional properties to recombinant receptors and may be particularly important in mediating tonic inhibition. Mice that lack the delta subunit have been produced by gene-targeting technology, and these mice were studied with immunohistochemical and immunoblot methods to determine whether changes in GABA(A) receptors were limited to deletion of the delta subunit or whether alterations in other GABA(A) receptor subunits were also present in the delta subunit knockout (delta-/-) mice. Immunohistochemical studies of wild-type mice confirmed the restricted distribution of the delta subunit in the forebrain. Regions with moderate to high levels of delta subunit expression included thalamic relay nuclei, caudate putamen, molecular layer of the dentate gyrus, and outer layers of the cerebral cortex. Virtually no delta subunit labeling was evident in adjacent regions, such as the thalamic reticular nucleus, hypothalamus, and globus pallidus. Comparisons of the expression of other subunits in delta-/- and wild-type mice demonstrated substantial changes in the alpha4 and gamma2 subunits of the GABA(A) receptor in the delta-/- mice. gamma2 Subunit expression was increased, whereas alpha4 subunit expression was decreased in delta-/- mice. Importantly, alterations of both the alpha4 and the gamma2 subunits were confined primarily to brain regions that normally expressed the delta subunit. This suggests that the additional subunit changes are directly linked to loss of the delta subunit and could reflect local changes in subunit composition and function of GABA(A) receptors in delta-/- mice. PMID- 11932937 TI - GABAergic innervation of alpha type II calcium/calmodulin-dependent protein kinase immunoreactive pyramidal neurons in the rat basolateral amygdala. AB - Although calcium/calmodulin-dependent protein kinase II (CaMK) has been shown to play a critical role in long-term potentiation (LTP) and emotional learning mediated by the basolateral amygdala, little is known about its cellular localization in this region. We have utilized immunohistochemical methods to study the neuronal localization of CaMK, and its relationship to gamma aminobutyric acid (GABA)-ergic structures, in the rat basolateral amygdala (ABL). Light microscopic observations revealed dense CaMK staining in the ABL. Although the cell bodies and proximal dendrites of virtually every pyramidal cell appeared to be CaMK(+), the cell bodies of small nonpyramidal neurons were always unstained. Dual localization of CaMK and GABA immunoreactivity with confocal immunofluorescence microscopy revealed that CaMK and GABA were found in different neuronal populations in the ABL. CaMK was contained only in pyramidal neurons; GABA was contained only in nonpyramidal cells. At the ultrastructural level, it was found that CaMK was localized to pyramidal cell bodies, thick proximal dendrites, thin distal dendrites, most dendritic spines, axon initial segments, and axon terminals forming asymmetrical synapses. These findings suggest that all portions of labeled pyramidal cells, with the exception of some dendritic spines, can exhibit CaMK immunoreactivity. By using a dual immunoperoxidase/immunogold silver procedure at the ultrastructural level, GABA(+) axon terminals were seen to innervate all CaMK(+) postsynaptic domains, including cell bodies (22%), thick (>1 microm) dendrites (34%), thin (<1 microm) dendrites (22%), dendritic spines (17%), and axon initial segments (5%). These findings indicate that CaMK is a useful marker for pyramidal neurons in ultrastructural studies of ABL synaptology and that the activity of pyramidal neurons in the ABL is tightly controlled by a high density of GABAergic terminals that target all postsynaptic domains of pyramidal neurons. PMID- 11932938 TI - Distribution and synaptic connectivity of neuropeptide Y-immunoreactive amacrine cells in the rat retina. AB - Neuropeptide Y (NPY) is a potent bioactive peptide that is widely expressed in the nervous system, including the retina. Here we show that specific NPY immunoreactivity was localized to amacrine and displaced amacrine cells in the rat retina. Immunoreactive cells had a regular distribution across the retina and an overall cell density of 280 cells/mm(2) in the inner nuclear layer (INL) and 90 cells/mm(2) in the ganglion cell layer (GCL). In the INL, most immunoreactive cells were characterized by small cell bodies and fine processes that appeared to ramify primarily in stratum 1 of the inner plexiform layer (IPL). A few cells in the INL also ramified in stratum 3 of the IPL. In the GCL, small to medium immunoreactive cells appeared to ramify primarily in stratum 5 of the IPL. A few immunoreactive processes, originating from somata in the INL and processes in the IPL, ramified in the OPL. NPY-immunoreactive cells contained GABA immunoreactivity, and some amacrine cells also contained tyrosine hydroxylase immunoreactivity. NPY-immunostained processes were most frequently presynaptic to nonimmunostained amacrine and ganglion cell processes and postsynaptic to nonimmunostained amacrine cell processes and cone bipolar cell axonal terminals. These findings indicate that NPY immunoreactivity is present in two populations of amacrine cells, one located in the INL and the other in the GCL, and that these cells mainly form synaptic contacts with other amacrine cells. These observations suggest that NPY-immunoreactive cells participate in multiple circuits mediating visual information processing in the inner retina. PMID- 11932939 TI - Corticotropin-releasing hormone-synthesizing neurons of the human hypothalamus receive neuropeptide Y-immunoreactive innervation from neurons residing primarily outside the infundibular nucleus. AB - Immunohistochemical single- and double-labeling studies were performed on the hypothalami of postmortem human brains to elucidate the distribution of corticotropin-releasing hormone (CRH)-immunoreactive (IR) neuronal elements and their interaction with the neuropeptide Y (NPY)-ergic neuronal system. The great majority of CRH-IR perikarya were found in the paraventricular nucleus (PVN), whereas a considerable number of CRH-IR neurons were also observed in the periventricular and infundibular nuclei. The dorsomedial nucleus and the perifornical region contained only scattered CRH-IR neurons. Dense CRH-IR fiber networks were found throughout the hypothalamus. However, the medial preoptic, the dorsolateral part of the supraoptic, the suprachiasmatic, the ventromedial, and the different mammillary nuclei showed a relative paucity of fibers. The terminal fields of NPY-IR axons overlapped the distribution of CRH-IR neurons in the hypothalamus. NPY-IR axon varicosities were juxtaposed to both dendrites and perikarya of the majority of CRH-IR neurons residing in the paraventricular, periventricular, and infundibular nuclei. These neurons were frequently contacted by multiple NPY axons that either formed baskets around their perikarya or completely ensheathed the emanating CRH dendrites. Because NPY and agouti-related protein (AGRP) are co-contained in neurons of the human infundibular nucleus, we used AGRP as a marker of NPY fibers originating exclusively from the infundibular nucleus. Only a small proportion of CRH neurons in the PVN was contacted by AGRP IR axon varicosities, suggesting that NPY-IR innervation of CRH neurons in the PVN derive mainly from regions outside the infundibular nucleus. The present morphological findings support the view that NPY regulates the CRH system of the human hypothalamus and therefore at least some of the effects of NPY on metabolic, autonomic, and endocrine functions may be mediated through CRH. PMID- 11932940 TI - Polysialic acid and the formation of oculomotor synapses on chick ciliary neurons. AB - The polysialic acid (PSA) moiety of the neural cell adhesion molecule (NCAM) participates in a variety of developmental processes, including axonal guidance and cell migration. PSA's function in these contexts stems from its ability to reduce cell interactions. The present study examines the regulation of PSA expression during formation of the calyciform synapse by the oculomotor axons on chick ciliary neurons. Prior to synaptogenesis, PSA is abundantly and uniformly expressed on the surface of the ciliary neuron body. However, at the time synaptic bonds start to form, as reflected in the localized accumulation of synaptic vesicles, PSA is lost from the point of synaptic contact. Thereafter, PSA is progressively lost from the ciliary neuron surface as the calyx grows. The dense mats of pseudodendritic-like somatic spines, which extend from the postsynaptic cell body, form an exception. These spines, which are known to undergo morphological remodeling, retain PSA expression until the end of embryogenesis. The experimental removal of PSA did not affect synaptogenesis itself, in that no significant changes were observed in the surface covered by the calyx, the number of spine aggregates, the size of acetylcholine receptor clusters, the cell surface area covered by these receptors, or the ultrastructure of the calyx, spine mats, and active zones. Together, these observations suggest that the synapse eliminates PSA as a part of its normal development and that the loss of PSA from the site of axon-target interaction may serve to stabilize structures formed during synaptogenesis. PMID- 11932941 TI - Axonopathy, tau abnormalities, and dyskinesia, but no neurofibrillary tangles in p25-transgenic mice. AB - Neurofibrillary tangles, one of the pathologic hallmarks of Alzheimer's disease (AD), are composed of abnormally polymerized tau protein. The hyperphosphorylation of tau alters its normal cellular function and is thought to promote the formation of neurofibrillary tangles. Growing evidence suggests that cyclin-dependent kinase 5 (cdk5) plays a role in tau phosphorylation, but the function of the enzyme in tangle formation remains uncertain. In AD, cdk5 is constitutively activated by p25, a highly stable, 25kD protein thought to be increased in the AD brain. To test the hypothesis that p25/cdk5 interactions promote neurofibrillary pathology, we created transgenic mouse lines that overexpress the human p25 protein specifically in neurons. Mice with high transgenic p25 expression have augmented cdk5 activity and develop severe hindlimb semiparalysis and mild forelimb dyskinesia beginning at approximately 3 months of age. Immunohistochemical and ultrastructural analyses showed widespread axonal degeneration with focal accumulation of tau in various regions of the brain and, to a lesser extent, the spinal cord. However, there was no evidence of neurofibrillary tangles in neuronal somata or axons, nor were paired helical filaments evident ultrastructurally. These studies confirm that p25 overexpression can lead to tau abnormalities and axonal degeneration in vivo but do not support the hypothesis that p25-related induction of cdk5 is a primary event in the genesis of neurofibrillary tangles. PMID- 11932942 TI - Differentiation and maturation of zebrafish dorsal root and sympathetic ganglion neurons. AB - The trunk neural crest of vertebrate embryos gives rise to dorsal root ganglion (DRG) sensory neurons and autonomic sympathetic neurons, among other derivatives. We have examined the development of DRG and sympathetic neurons during development in the zebrafish. We found that sensory neurons differentiate rapidly and that their overt neuronal differentiation significantly precedes that of sympathetic neurons in the trunk. Sympathetic neurons in different regions differentiate at different times. The most rostral population, which we call the cervical ganglion, differentiates several days before trunk sympathetic neurons. After undergoing overt neuronal differentiation, sympathetic neurons subsequently express the adrenergic differentiation markers dopamine beta-hydroxylase and tyrosine hydroxylase. A second population of adrenergic nonneuronal cells initially localized with cervical sympathetic neurons appears to represent adrenal chromaffin cells. In more mature fish, these cells were present in clusters within the kidneys. Individual DRG and sympathetic ganglia initially contain few neurons. However, the number of neurons in DRG and sympathetic ganglia increases continuously at least up to 4 weeks of age. Analysis of phosphohistone H3 expression and bromodeoxyuridine incorporation studies suggests that the increases in DRG and sympathetic ganglion neuronal cell number are due wholly or in part to the division of neuronal cells within the ganglia. PMID- 11932943 TI - Extent of retinal ganglion cell death in the frog Litoria moorei after optic nerve regeneration induced by lesions of different sizes. AB - Some amphibian retinal ganglion cells die during optic nerve regeneration. Here we have investigated whether ganglion cell death in the frog Litoria moorei is associated with the lesion site. For one experimental series, the optic nerve lesion extended for 0.15 mm; in the other, it extended for 1.5 mm. The extent of ganglion cell death was estimated from cresyl violet-stained whole mounts at 24 weeks post lesion. In other animals, individual regenerating axons were visualised in the optic nerve by horseradish peroxidase (HRP) labelling from 1 day to 24 weeks post lesion; counterstaining with cresyl violet allowed examination of cells that repopulated the lesion site. Ganglion cell numbers fell significantly more after an extensive than after a localised lesion, long-term losses being 50% and 34%, respectively (P < 0.05). Regenerating axons were delayed in their passage across the cell-poor extensive lesion compared with the relatively cell-rich localised lesion. The differing rates of regeneration between series were matched by greater delay after extensive lesion in the return of visually guided behaviour as assessed by optokinetic horizontal head nystagmus. We suggest that delays in regeneration after an extensive lesion exacerbate ganglion cell death, indicating that conditions within the lesion are associated with the death of some ganglion cells. PMID- 11932944 TI - Estrogen receptor-beta immunoreactivity in the midbrain of adult rats: regional, subregional, and cellular localization in the A10, A9, and A8 dopamine cell groups. AB - Estrogen modulates dopamine synthesis, release, and metabolism in corticolimbic and striatal targets of midbrain dopamine neurons. The relevant sites of receptor mediated action, however, had been elusive, because all available evidence suggested a paucity of intracellular estrogen receptors in the A8, A9, and A10 dopamine regions and their afferent targets. More recent evidence of a relative abundance of the beta isoform of the estrogen receptor (ER) in the substantia nigra and ventral tegmental area (VTA), however, suggests that this newly described receptor may be important in estrogen's stimulation of midbrain DA systems. It is unknown, however, precisely how ERbeta is distributed with respect to the functionally and neurochemically diverse cell populations of the ventral midbrain. To address these issues, this study used single- and double-label immunocytochemistry to detail the regional, subregional, and cellular distributions of ERbeta immunoreactivity in and around midbrain dopamine containing cell groups in hormonally intact adult male and female rats. These analyses revealed that ERbeta-immunoreactive nuclei were found only in neurons, more specifically, within subsets of both dopaminergic and nondopaminergic neurons in the dorsal VTA, the parabrachial pigmented nucleus, the substantia nigra pars lateralis, the retrorubral fields, and to a lesser extent the linear midline nuclei. These regional and cellular receptor distributions thus place the ERbeta isoform in anatomical register with midbrain dopamine systems known to participate in a spectrum of motor, cognitive, and affective functions. PMID- 11932945 TI - Molecular phylogenetic perspective on evolution of lizards of the Anolis grahami series. AB - We report the results of phylogenetic analyses of 1447 bases of mitochondrial DNA sequence for 21 populations representing seven species of the Anolis grahami series (A. conspersus, A. garmani, A. grahami, A. lineatopus, A. opalinus, A. reconditus, and A. valencienni), six of which occur on Jamaica. These data include 705 characters that are phylogenetically informative according to parsimony. A parsimony analysis of these data combined with previously published allozymic data yields a single most parsimonious tree with strong support for monophyly of the A. grahami series, the sister-group relationship between Anolis lineatopus and A. reconditus and a clade composed of Anolis garmani, A. grahami, and A. opalinus. Based on DNA data alone, A. conspersus is nested within A. grahami. Haplotypes sampled from geographic populations of A. grahami, A. lineatopus, and A. opalinus are highly divergent (approximately 12-15% sequence difference on average for each species) and show similar phylogeographic patterns, suggesting that each of these currently recognized species may be a complex of species. Anolis valencienni also shows high sequence divergence among haplotypes from different geographic populations (approximately 8% sequence difference) and may contain cryptic species. Divergence among haplotypes within A. garmani is substantially lower (approximately 3% sequence difference), and phylogeographic patterns are significantly different from those observed in A. grahami, A. lineatopus and A. opalinus. PMID- 11932946 TI - Identification and functional characterization of hypoxia-inducible factor 2alpha from the estuarine teleost, Fundulus heteroclitus: interaction of HIF-2alpha with two ARNT2 splice variants. AB - The hypoxia-inducible factors (HIFs) are dimeric transcription factors that mediate changes in gene expression during adaptation of animals to oxygen stress. Both alpha (HIFalpha) and beta (ARNT) subunits are members of the basic helix loop-helix/Per-ARNT-Sim family of proteins. Mammals have at least three different HIF-alpha subunits, paralogous proteins expressed in tissue-specific fashion (HIF 1alpha, HIF-2alpha, and HIF-3alpha). However, the diversity and functional properties of teleost HIFs are poorly understood. In efforts to characterize mechanisms of hypoxia adaptation in estuarine fish, we have isolated cDNAs encoding HIF subunits from Fundulus heteroclitus (Atlantic killifish or mummichog), including a HIF-2alpha homolog and ARNT2alt, a splice variant of ARNT2 that contains an additional exon encoding 16 amino acids near the amino terminus. HIF-2alpha protein synthesized in vitro binds cognate DNA elements in concert with either Fundulus ARNT2 splice variant or murine ARNT1. HIF-2alpha, ARNT2, and ARNT2alt mRNAs are expressed in all organs examined. The HIF-2alpha cDNA encodes a protein of 96.4 kDa, sharing 53-54% identity with mammalian and avian orthologs. The oxygen-dependent degradation domain, however, exhibits substantial divergence from well-conserved mammalian sequences, suggesting the possibility of important functional differences, perhaps in the sensitivity to induction of activity by hypoxia. Hypoxia-tolerant fishes such as F. heteroclitus represent a unique opportunity for the study of functional and evolutionary aspects of adaptation to hypoxia at the molecular, cellular, and organismal levels. This study extends the understanding of hypoxia signaling in fish, the evolution and diversity of HIF function, and the evolution of the PAS family of proteins. PMID- 11932947 TI - Hox cluster organization in the jawless vertebrate Petromyzon marinus. AB - Large-scale gene amplifications may have facilitated the evolution of morphological innovations that accompanied the origin of vertebrates. This hypothesis predicts that the genomes of extant jawless fish, scions of deeply branching vertebrate lineages, should bear a record of these events. Previous work suggests that nonvertebrate chordates have a single Hox cluster, but that gnathostome vertebrates have four or more Hox clusters. Did the duplication events that produced multiple vertebrate Hox clusters occur before or after the divergence of agnathan and gnathostome lineages? Can investigation of lamprey Hox clusters illuminate the origins of the four gnathostome Hox clusters? To approach these questions, we cloned and sequenced 13 Hox cluster genes from cDNA and genomic libraries in the lamprey, Petromyzon marinus. The results suggest that the lamprey has at least four Hox clusters and support the model that gnathostome Hox clusters arose by a two-round-no-cluster-loss mechanism, with tree topology [(AB)(CD)]. A three-round model, however, is not rigorously excluded by the data and, for this model, the tree topologies [(D(C(AB))] and [(C(D(AB))] are most parsimonious. Gene phylogenies suggest that at least one Hox cluster duplication occurred in the lamprey lineage after it diverged from the gnathostome lineage. The results argue against two or more rounds of duplication before the divergence of agnathan and gnathostome vertebrates. If Hox clusters were duplicated in whole genome duplication events, then these data suggest that, at most, one whole genome duplication occurred before the evolution of vertebrate developmental innovations. PMID- 11932948 TI - Genomic analysis of Hox clusters in the sea lamprey Petromyzon marinus. AB - The sea lamprey Petromyzon marinus is among the most primitive of extant vertebrates. We are interested in the organization of its Hox gene clusters, because, as a close relative of the gnathostomes, this information would help to infer Hox cluster organization at the base of the gnathostome radiation. We have partially mapped the P. marinus Hox clusters using phage, cosmid, and P1 artificial chromosome libraries. Complete homeobox sequences were obtained for the 22 Hox genes recovered in the genomic library screens and analyzed for cognate group identity. We estimate that the clusters are somewhat larger than those of mammals (roughly 140 kbp vs. 105 kbp) but much smaller than the single Hox cluster of the cephalochordate amphioxus (at more than 260 kb). We never obtained more than three genes from any single cognate group from the genomic library screens, although it is unlikely that our screen was exhaustive, and therefore conclude that P. marinus has a total of either three or four Hox clusters. We also identify four highly conserved non-coding sequence motifs shared with higher vertebrates in a genomic comparison of Hox 10 genes. PMID- 11932949 TI - A conserved family of doublesex-related genes from fishes. AB - The sex-determining gene Mab-3 of C. elegans and the doublesex gene of Drosophila each contain a common DM domain and share a similar role. Human doublesex-related gene DMRT1 also encodes a conserved DM-related DNA-binding domain. We present here the amplification of a broad range of DM domain sequences from three fish species using degenerate PCR. Our results reveal unexpected complexity of the DM domain gene family in vertebrates. PMID- 11932950 TI - Segment-specific expression of alkaline phosphatase in the Tubifex embryo requires DNA replication and mRNA synthesis. AB - During embryogenesis of the oligochaete annelid Tubifex, segments VII and VIII specifically express mesodermal alkaline phosphatase (ALP) activity in the ventrolateral region. In this study, using specific inhibitors, we examined whether this segment-specific expression of ALP activity depends on DNA replication and RNA transcription. BrdU-incorporation experiments showed that presumptive ALP-expressing cells undergo the last round of DNA replication at 12 24 hr prior to emergence of ALP activity. When this DNA replication was inhibited by aphidicolin, ALP development was completely abrogated in the ventrolateral mesoderm. Similar inhibition of ALP development was also observed in alpha amanitin-injected embryos. While injection of alpha-amanitin at 24 hr prior to the emergence of ALP activity exerted inhibitory effects on ALP development, injection at 14 hr was no longer effective. In contrast, ALP activity developed normally in cytochalasin-D-treated embryos in which cytokinesis was prevented from occurring for 36 hs prior to appearance of ALP activity. These results suggest that the segment-specific development of ALP activity in the Tubifex embryo depends on DNA replication and mRNA transcription, both of which occur long before the emergence of ALP activity. PMID- 11932951 TI - Stimulation of nitric oxide synthase in cerebral cortex due to elevated partial pressures of oxygen: an oxidative stress response. AB - The purpose of this investigation was to determine the impact of elevated partial pressures of O(2) on the steady state concentration of nitric oxide ((*)NO) in the cerebral cortex. Rodents with implanted O(2)- and (*)NO-specific microelectrodes were exposed to O(2) at partial pressures from 0.2 to 2.8 atmospheres absolute (ATA) for up to 45 min. Elevations in (*)NO concentration occurred with all partial pressures above that of ambient air. In rats exposed to 2.8 ATA O(2) the increase was 692 +/- 73 nM (S.E., n = 5) over control. Changes were not associated with alterations in concentrations of nitric oxide synthase (NOS) enzymes. Based on studies with knock-out mice lacking genes for neuronal NOS (nNOS) or endothelial NOS (eNOS), nNOS activity contributed over 90% to total (*)NO elevation due to hyperoxia. Immunoprecipitation studies indicated that hyperoxia doubles the amount of nNOS associated with the molecular chaperone, heat shock protein 90 (Hsp90). Both (*)NO elevations and the association between nNOS and Hsp90 were inhibited in rats infused with superoxide dismutase. Elevations of (*)NO were also inhibited by treatment with the relatively specific nNOS inhibitor, 7 nitroindazole, by the ansamycin antibiotics herbimycin and geldanamycin, by the antioxidant N-acetylcysteine, by the calcium channel blocker nimodipine, and by the N-methyl-D-aspartate inhibitor, MK 801. Hyperoxia did not alter eNOS association with Hsp90, nor did it modify nNOS or eNOS associations with calmodulin, the magnitude of eNOS tyrosine phosphorylation, or nNOS phosphorylation via calmodulin kinase. Cerebral cortex blood flow, measured by laser Doppler flow probe, increased during hyperoxia and may be causally related to elevations of steady state (*)NO concentration. We conclude that hyperoxia causes an increase in (*)NO synthesis as part of a response to oxidative stress. Mechanisms for nNOS activation include augmentation in the association with Hsp90 and intracellular entry of calcium. PMID- 11932952 TI - Comparison of the effects of HGF, BDNF, CT-1, CNTF, and the branchial arches on the growth of embryonic cranial motor neurons. AB - In the developing embryo, axon growth and guidance depend on cues that include diffusible molecules. We have shown previously that the branchial arches and hepatocyte growth factor (HGF) are growth-promoting and chemoattractant for young embryonic cranial motor axons. HGF is produced in the branchial arches of the embryo, but a number of lines of evidence suggest that HGF is unlikely to be the only factor involved in the growth and guidance of these axons. Here we investigate whether other neurotrophic factors could be involved in the growth of young cranial motor neurons in explant cultures. We find that brain-derived neurotrophic factor (BDNF), ciliary neurotrophic factor (CNTF) and cardiotrophin 1 (CT-1) all promote the outgrowth of embryonic cranial motor neurons, while glial cell line-derived neurotrophic factor (GDNF) and neurotrophin-3 (NT-3) fail to affect outgrowth. We next examined whether HGF and the branchial arches had similar effects on motor neuron subpopulations at different axial levels. Our results show that HGF acts as a generalized rather than a specific neurotrophic factor and guidance cue for cranial motor neurons. Although the branchial arches also had general growth-promoting effects on all motor neuron subpopulations, they chemoattracted different axial levels differentially, with motor neurons from the caudal hindbrain showing the most striking response. PMID- 11932953 TI - The effects of social environment on adult neurogenesis in the female prairie vole. AB - In the mammalian brain, adult neurogenesis has been found to occur primarily in the subventricular zone (SVZ) and dentate gyrus of the hippocampus (DG) and to be influenced by both exogenous and endogenous factors. In the present study, we examined the effects of male exposure or social isolation on neurogenesis in adult female prairie voles (Microtus ochrogaster). Newly proliferated cells labeled by a cell proliferation marker, 5-bromo-2'-deoxyuridine (BrdU), were found in the SVZ and DG, as well as in other brain areas, such as the amygdala, hypothalamus, neocortex, and caudate/putamen. Two days of male exposure significantly increased the number of BrdU-labeled cells in the amygdala and hypothalamus in comparison to social isolation. Three weeks later, group differences in BrdU labeling generally persisted in the amygdala, whereas in the hypothalamus, the male-exposed animals had more BrdU-labeled cells than did the female-exposed animals. In the SVZ, 2 days of social isolation increased the number of BrdU-labeled cells compared to female exposure, but this difference was no longer present 3 weeks later. We have also found that the vast majority of the BrdU-labeled cells contained a neuronal marker, indicating neuronal phenotypes. Finally, group differences in the number of cells undergoing apoptosis were subtle and did not seem to account for the observed differences in BrdU labeling. Together, our data indicate that social environment affects neuron proliferation in a stimulus- and site-specific manner in adult female prairie voles. PMID- 11932954 TI - Respiratory units of motor production and song imitation in the zebra finch. AB - Juvenile male zebra finches (Taeniopygia guttata) learn a stereotyped song by imitating sounds from adult male tutors. Their song is composed of a series of syllables, which are separated by silent periods. How acoustic units of song are translated into respiratory and syringeal motor gestures during the song learning process is not well understood. To learn about the respiratory contribution to the imitation process, we recorded air sac pressure in 38 male zebra finches and compared the acoustic structures and air sac pressure patterns of similar syllables qualitatively and quantitatively. Acoustic syllables correspond to expiratory pressure pulses and most often (74%) entire syllables are copied using similar air sac pressure patterns. Even notes placed within different syllables are generated with similar air sac pressure patterns when only segments of syllables are copied (9%). A few of the similar syllables (17%) are generated with a modified pressure pattern, typically involving addition or deletion of an inspiration. The high similarity of pressure patterns for like syllables indicates that generation of particular sounds is constrained to a narrow range of air sac pressure conditions. Following presentation of stroboscope flashes, song was typically interrupted at the end of an expiratory pressure pulse, confirming that expirations and, therefore, syllables are the smallest unit of motor production of song. Silent periods, which separate syllables acoustically, are generated by switching from expiration to inspiration. Switching between respiratory phases, therefore, appears to play a dominant role in organizing the stereotyped motor program for song production. PMID- 11932955 TI - Changes in spatial memory mediated by experimental variation in food supply do not affect hippocampal anatomy in mountain chickadees (Poecile gambeli). AB - Earlier reports suggested that seasonal variation in food-caching behavior (caching intensity and cache retrieval accuracy) might correlate with morphological changes in the hippocampal formation, a brain structure thought to play a role in remembering cache locations. We demonstrated that changes in cache retrieval accuracy can also be triggered by experimental variation in food supply: captive mountain chickadees (Poecile gambeli) maintained on limited and unpredictable food supply were more accurate at recovering their caches and performed better on spatial memory tests than birds maintained on ad libitum food. In this study, we investigated whether these two treatment groups also differed in the volume and neuron number of the hippocampal formation. If variation in memory for food caches correlates with hippocampal size, then our birds with enhanced cache recovery and spatial memory performance should have larger hippocampal volumes and total neuron numbers. Contrary to this prediction we found no significant differences in volume or total neuron number of the hippocampal formation between the two treatment groups. Our results therefore indicate that changes in food-caching behavior and spatial memory performance, as mediated by experimental variations in food supply, are not necessarily accompanied by morphological changes in volume or neuron number of the hippocampal formation in fully developed, experienced food-caching birds. PMID- 11932956 TI - Developmental and hormonal regulation of NR2A mRNA in forebrain regions controlling avian vocal learning. AB - Developmental changes in the composition of NMDA receptors can alter receptor physiology as well as intracellular signal transduction cascades, potentially shifting thresholds for neural and behavioral plasticity. During song learning in zebra finches, NMDAR currents become faster, and transcripts for the modulatory NR2B subunit of this receptor decrease in lMAN, a region in which NMDAR activation is critical for vocal learning. Using in situ hybridization, we found that NR2A transcripts change reciprocally, increasing significantly in both lMAN (59%) and in another song region, Area X (38%), between posthatch day (PHD) 20 and 40, but not changing further at PHD60 or 80. In adjacent areas not associated with song learning, NR2A mRNA did not change between PHD20-80. Although early song deprivation (which extends the sensitive period for song learning) delays changes in NR2B gene expression and NMDAR physiology within the lMAN, it did not alter NR2A mRNA levels measured at PHD40, 45, or 60. Early testosterone (T) treatment, which disrupts vocal development and accelerates the maturation of both NR2B levels and NMDAR physiology in lMAN, also significantly increased NR2A transcripts measured at PHD35 in lMAN. In Area X, a similar effect of T approached significance. Together with our previous studies, these results show that in a pathway critical for vocal plasticity, the ratio of NR2A:NR2B mRNA rises abruptly early during the sensitive period for song learning. Furthermore, androgen regulation of NMDAR gene expression may alter thresholds for experience dependent synaptic change. PMID- 11932957 TI - Influence of ethanol on neonatal cerebellum of BDNF gene-deleted animals: analyses of effects on Purkinje cells, apoptosis-related proteins, and endogenous antioxidants. AB - The sensitivity of the developing central nervous system (CNS) to the deleterious effects of ethanol has been well documented, with exposure leading to a wide array of CNS abnormalities. Certain CNS regions are susceptible to ethanol during well-defined critical periods. In the neonatal rodent cerebellum, a profound loss of Purkinje cells is found when ethanol is administered early in the postnatal period [on postnatal days 4 or 5 (P4-5)], while this neuronal population is much less vulnerable to similar ethanol insult slightly later in the postnatal period (P7-9). Prior studies have shown that neurotrophic factors (NTFs) can be altered by ethanol exposure, and both in vitro and in vivo studies have provided evidence that such substances have the potential to protect against ethanol neurotoxicity. In the present study, it was hypothesized that depletion of an NTF shown to be important to cerebellar development would exacerbate ethanol-related effects within this region, when administration was confined to a normally ethanol resistant ontogenetic period. For this study, brain-derived neurotrophic factor (BDNF) gene-deleted ("knockout") and wild-type mice were exposed to ethanol via vapor inhalation or to control conditions during the normally ethanol-resistant period (P7 and P8). Two hours after termination of exposure on P8, analyses were made of body weight, crown-rump length, and brain weight. In subsequent investigations, the number and density of Purkinje cells and the volume of cerebellar lobule I were determined, and the expression of anti- and pro apoptotic proteins and the activities of endogenous antioxidants were assessed. It was found that the BDNF knockouts were significantly smaller than the wild type animals, with smaller brain weights. Purkinje cell number and density was reduced in ethanol-treated knockout, but not wild-type animals, and the volume of lobule I was significantly decreased in the gene-deleted animals compared to wild types, but was not further affected by ethanol treatment. The loss of Purkinje cells in the BDNF knockouts was accompanied by decreases in anti-apoptotic Bcl-xl and in phosphorylated (and hence inactivated) pro-apoptotic Bad, and reduced activity of the antioxidant glutathione reductase, while the antioxidant catalase was increased by ethanol treatment in this genotype. In the wild-type animals, anti-apoptotic Bcl-2 was decreased by ethanol treatment, but the pro-apoptotic c Jun N-terminal kinase (JNK) was markedly diminished by ethanol exposure, while the activity of the protective antioxidant superoxide dismutase (SOD) was significantly enhanced. These results suggest that neurotrophic factors have the capacity to protect against ethanol neurotoxicity, perhaps by regulation of expression of molecules critical to neuronal survival such as elements of the apoptosis cascade and protective antioxidants. PMID- 11932958 TI - A study of streptomycin blood level information of patients undergoing hemodialysis. AB - Pharmacokinetic feature of streptomycin (SM) was investigated before and during hemodialysis (HD) in four patients with renal failure undergoing HD. SM concentrations were assayed by using TDX SM KIT (DAINABOT). Patients received 10 mg/kg of SM by intramuscular injection before HD. Pharmacokinetic parameters of SM intramuscular injection before HD were k(a)=2.38+/-0.53 h(-1); k(e)=0.0130+/ 0.0025 h(-1); V(d)=0.313+/-0.026l/kg and t(1/2)=55.6+/-10.4 h. The maximum concentration (C(max)) of SM was observed at about 2 h after the SM administration and the mean serum concentration of SM was 30.4 microg/ml; even 4 h after the SM injection, the concentration still remained in a range over 30 microg/ml. The data suggest that a possible toxicity might have appeared in the patients. During the hemodialysis an average t(1/2) value was 3.32 h. This value is close to the value of a healthy person. The k(e) value of patient A during the hemodialysis became 24 times as large as that observed before the hemodialysis. On the average it was 17 times as large as that observed before the hemodialysis. Thus, it was found that the values of pharmacokinetics parameters such as k(e) and t(1/2) during the hemodialysis were similar to those of a healthy person, although there are some variations. PMID- 11932959 TI - Minimal effect of ketoconazole on cyclosporine (SangCyA) oral absorption and first-pass metabolism in rats: evidence of a significant vehicle effect on SangCyA absorption. AB - The current work evaluated the effect of the CYP3A inhibitor ketoconazole on the oral absorption and first-pass metabolism of cyclosporine administered as the SangCyA formulation. Groups of 6 male Sprague-Dawley rats were administered SangCyA (5 and 15 mg/kg) by oral gavage alone and with ketoconazole (30 mg/kg). Blood cyclosporine levels were measured over 6 h, encompassing the cyclosporine absorption window. A significant vehicle effect on SangCyA absorption was observed. Comparing a 15 mg/kg dose, cyclosporine C(max) (mean+/-SD 1.12+/-0.16 microg/ml) and AUC(0-6) (5.34+/-0.71 microg h/ml) were 50% lower when propylene glycol was used as gavage vehicle instead of saline (2.19+/-0.94 microg/ml and 9.52+/-2.52 microg h/ml, respectively). Coefficients-of-variation for these parameters were halved in the propylene glycol vehicle however T(max) was unaffected. Ketoconazole increased cyclosporine C(max) and AUC(0-6) by 50-60%, regardless of the vehicle or the cyclosporine dose, without altering T(max) (2-3 h). The small effect of ketoconazole suggests that CYP3A-mediated intestinal and first-pass hepatic metabolism are minor determinants of cyclosporine oral bioavailability in rats. PMID- 11932960 TI - Absence of clinically relevant drug interactions following simultaneous administration of didanosine-encapsulated, enteric-coated bead formulation with either itraconazole or fluconazole. AB - This open-label, two-way crossover study was undertaken to determine whether the enteric formulation of didanosine influences the pharmacokinetics of itraconazole or fluconazole, two agents frequently used to treat fungal infections that occur with HIV infection, and whose bioavailability may be influenced by changes in gastric pH. Healthy subjects were randomized to Treatment A (200-mg itraconazole or 200-mg fluconazole) or Treatment B (same dose of itraconazole or fluconazole with 400 mg of didanosine as an encapsulated, enteric-coated bead formulation). In the itraconazole study, a lack of interaction was concluded if the 90% confidence interval (CI) of the ratio of the geometric means of log-transformed C(max) and AUC(0-T) values of itraconazole and hydroxyitraconazole, the active metabolite of itraconazole, were contained entirely between 0.75 and 1.33. In the fluconazole study, the equivalence interval for C(max) and AUC(0-T) was 0.80 1.25. The data showed that for itraconazole the point estimate and 90% CI of the ratios of C(max) and AUC(0-T) values were 0.98 (0.79, 1.20) and 0.88 (0.71, 1.09), respectively; for hydroxyitraconazole the respective values were 0.91 (0.76, 1.08) and 0.85 (0.68, 1.06). In the fluconazole study, the point estimate and 90% CI of the ratios of C(max) and AUC(0-T) values were 0.98 (0.93, 1.03) and 1.01 (0.99, 1.03), respectively. The T(max) for itraconazole, hydroxyitraconazole, and fluconazole were similar between treatments. Both studies indicated a lack of clinically significant interactions of the didanosine formulation with itraconazole or fluconazole. These results showed that the encapsulated, enteric-coated bead formulation of didanosine can be concomitantly administered with drugs, such as the azole antifungal agents, whose bioavailability may be influenced by interaction with antacids. PMID- 11932961 TI - Disposition and receptor-site binding of (125)I-EGF after topical administration to skin wounds. AB - The rhEGF topical delivery systems have been hindered by a number of shortcomings which have led to the search of new development strategies. In this study we report the evaluation of cumulative profiles of 10, 5 and 1 microg/ml solutions of (125)I-rhEGF, in a rat full-thickness skin wound model, as well as the drug induced modulation in the expression of the EGF receptor after lesion. The tissue associated radioactivity, expressed as the percentage of the dose administered per grams of tissue (%D/g), peaks at 2 h after administration of all doses. (125)I-rhEGF degraded species were detected chromatographically, but no diffusion of the peptide to the surrounding skin was documented. Despite the dose, the EGF receptor expression was increased within 2 h after wounding, followed by a slow decline up to 12 h below baseline. Twelve hours after punch, differences were evident between all treated groups and control. These results demonstrate that (125)I-rhEGF saline solutions are rapidly cleared from application sites, probably by protease-driven cleavage and receptor-mediated endocytosis. Finally, we must be aware that the results herein discussed should be taken into account during the drug delivery system design in order to guarantee the necessary steady state rhEGF levels upon wound healing process. PMID- 11932962 TI - Influence of fluconazole on the pharmacokinetics of omeprazole in healthy volunteers. AB - Influence of fluconazole on the pharmacokinetics of omeprazole was evaluated by single oral administration of omeprazole capsule 20 mg (control group), or single oral administration of fluconazole capsule, 100 mg, and omeprazole, 20 mg, after 4 days of daily oral administration of fluconazole, 100 mg (treated group), to 18 healthy male volunteers. Omeprazole is extensively metabolized in the liver through 5-hydroxylation and sulfoxidation reactions catalyzed predominantly by CYP2C19 and CYP3A4, respectively. Fluconazole is a potent competitive inhibitor of CYP2C19 and a weak inhibitor of CYP3A4. In treated group, the area under the plasma concentration-time curve of omeprazole from time zero to time infinity (AUC) was significantly greater (3090 vs 491 ng h/ml), terminal half-life of omeprazole was significantly longer (2.59 vs 0.85 h), and peak plasma concentration of omeprazole (C(max)) was significantly higher (746 vs 311 ng/ml) than that in control group. The greater AUC and higher C(max) in treated group could be due to inhibition of omeprazole metabolism by fluconazole. PMID- 11932963 TI - Pharmacokinetics and dose proportionality of BMS-204352 after intravenous administration to dogs. AB - BMS-204352 is a novel maxi-K channel opener that is being developed for the treatment for stroke. The current study was designed to evaluate the dose proportionality and pharmacokinetics of BMS-204352 in dogs. In an open, three-way crossover study, three beagle dogs received a single intravenous dose of BMS 204352 as a 6-min infusion into the femoral vein at 0.4, 0.9, and 2.0 mg/kg dose levels. There was at least a 1-week washout period between treatments. Serial blood samples were collected for up to 32 h post dose and plasma samples were analyzed for the concentrations of intact BMS-204352 using a validated liquid chromatographic mass spectrometric (LC/MS) method. Pharmacokinetic analysis was performed using a non-compartmental method. Results indicated that peak BMS 204352 concentrations (C(max)) and area under the plasma concentration-time curves (AUC) values increased in a dose proportional manner. Mean residence time (MRT, 18.2-21.9 h) and elimination half-life (T(half), 13.5-17 h) did not change with dose. There was no dose dependency in the mean BMS-204352 total body clearance (CLT, 134-158 ml/h/kg) and mean steady state volume of distribution (VSS, 2839-3291 ml/kg). The high VSS value indicated that BMS-204352 was distributed extensively in the extravascular tissues. In conclusion, BMS-204352 exhibits linear pharmacokinetics over the dose range tested (0.4-2 mg/kg). PMID- 11932964 TI - The problem of fibromyalgia. PMID- 11932965 TI - Diabetic and nondiabetic lumbosacral radiculoplexus neuropathies: new insights into pathophysiology and treatment. AB - Diabetic lumbosacral radiculoplexus neuropathy (DLRPN) (also called diabetic amyotrophy) is a well-recognized subacute, painful, asymmetric lower-limb neuropathy that is associated with weight loss and type II diabetes mellitus. Nondiabetic lumbosacral radiculoplexus neuropathy (LRPN) has received less attention. Comparison of large cohorts with DLRPN and LRPN demonstrated that age at onset, course, type and distribution of symptoms and impairments, laboratory findings, and outcomes are similar. Both conditions are lumbosacral radiculoplexus neuropathies that are associated with weight loss and begin focally with pain but that evolve into widespread, bilateral paralytic disorders. Although both are monophasic illnesses, patients have prolonged morbidity from pain and weakness, and many patients become wheelchair-dependent. Although motor predominant, there is unequivocal evidence that autonomic and sensory nerves are also involved. Cutaneous nerves from patients with DLRPN and LRPN show pathological evidence of ischemic injury (multifocal fiber loss, perineurial thickening and degeneration, neovascularization, microfasciculation, and swollen axons with accumulated organelles) and microvasculitis (mural and perivascular inflammation, separation and fragmentation of mural smooth muscle layers of microvessels and hemosiderin-laden macrophages). Controlled trials with immune modulating therapies in DLRPN are in progress, and preliminary data suggest that such therapy may be beneficial in LRPN. It is likely that DLRPN and LRPN are immune-mediated neuropathies that should be separated from chronic inflammatory demyelinating polyneuropathy and from systemic necrotizing vasculitis. PMID- 11932966 TI - Low-level exposures to organophosphorus esters and peripheral nerve function. AB - This review evaluates the epidemiological and clinical evidence linking low-level and prolonged exposures to organophosphorus esters, used as insecticides or nerve agents, to peripheral nerve dysfunction. The clinical effects of large doses of these chemicals-including the cholinergic syndrome, the intermediate syndrome, and the delayed polyneuropathy-are all well established and are summarized. Based on these clinical observations and experimental studies, dose-effect relationships indicate that peripheral neuropathy always develops after cholinergic toxicity. However, several studies have suggested that this relationship may be different after low-level prolonged exposures, as, for instance, those experienced by Gulf War veterans and British sheep farmers, thereby leading to the development of peripheral neuropathy without preceding cholinergic toxicity. A critical assessment of these studies, involving subjects with either current or past exposures, indicates that changes in peripheral nerve function were mild, inconsistent, and unexplained and that most studies lack exposure data. Suggestions made about individual hypersusceptibility to delayed polyneuropathy lack support. It is concluded that there is no evidence of peripheral nerve dysfunction caused by low-level prolonged exposures to organophosphate insecticides or nerve agents. PMID- 11932967 TI - Age-related change in motor unit activation strategy in force production: a mechanomyographic investigation. AB - This study was designed to examine the effect of increasing age on the recruitment and activation of motor units (MU) in the biceps brachii muscle, using the mechanomyogram (MMG)/force relationship during isometric ramp contractions. The relationships between the root mean squared amplitude (RMS) and mean power frequency (MPF) of the MMG and relative force (% MVC) in the elderly (male, n = 10, age = 69.8 +/- 4.7 years, mean +/- SD) were markedly different from those in the young group (male, n = 15, age = 22.7 +/- 1.8 years). In elderly individuals, the RMS increased progressively with force up to 57.6 +/- 3.4% MVC, when a brief rapid increase was followed by a stable trend beyond 63.6 +/- 3.7% MVC. The MPF increased slowly up to 59.4 +/- 2.3% MVC; after a temporary reduction from 59.4% to 64.3 +/- 2.0% MVC, it then increased progressively again. In conjunction with absolute force (F(abs)), both the RMS and MPF in the elderly were smaller than those in the young group throughout the submaximal levels of force exerted. The results reflect an alteration in MU activation strategy, with a predominant role for MUs with slow-twitch fibers and an effective fused tetanus induced at lower firing rate of the MUs, resulting from age-related neuromuscular changes. PMID- 11932968 TI - Novel COL6A1 splicing mutation in a family affected by mild Bethlem myopathy. AB - Bethlem myopathy is an early-onset benign myopathy characterized by proximal muscular weakness and multiple flexion contractures. It is a dominantly inherited disorder associated with mutations in the three COL6 genes encoding type VI collagen. We detected a g-->a substitution at +1 position of COL6A1 intron 3 in a four-generation Italian family affected by a mild form of Bethlem myopathy. The mutation results in the activation of a cryptic splice donor site at the 3' end of exon 3, leading to the loss of 66 nucleotides and an "in-frame" deletion of 22 amino acids in the NH2-domain. Molecular analysis on fibroblasts of the propositus showed that the mutated mRNA was present and stable, but the mutated protein could not be detected. Western blot and immunofluorescence analyses showed a decreased level of collagen VI synthesis and deposition in fibroblasts of the propositus. Together, the results suggest that the mutated protein was highly unstable and rapidly degraded, and that the mild phenotype was caused by a reduced amount of normal collagen VI microfibrils. In addition, we demonstrated that lymphocytes can be used for the first mutation screening analysis of patients with Bethlem myopathy. PMID- 11932969 TI - Longitudinal study of functional spinal alpha motor neuron loss in amyotrophic lateral sclerosis. AB - Using a microstimulation technique for obtaining motor unit number estimates (MUNEs) of the hypothenar and extensor digitorum brevis (EDB) muscles, we performed a longitudinal study on the natural course of change in the clinical rating scale (Appel score) and of loss of functional spinal alpha motor neurons in amyotrophic lateral sclerosis. The Appel score increased to about 150% of normal at 12 months after onset, about 225% at 18 months after onset, and about 370% at 24 months after onset. By contrast, MUNEs decreased to about 27% of normal at 12 months after onset, about 12% at 18 months after onset, and about 5% at 24 months after onset. The relative merits of these different approaches in detecting changes in the disease process in its early phase are discussed. PMID- 11932970 TI - Vibration injury damages arterial endothelial cells. AB - Prolonged exposure to hand-transmitted vibration can cause debilitating neural and vascular dysfunction in humans. It is unclear whether the pathophysiology involves simultaneous or sequential injury of arteries and nerves. The mechanism of vibration injury was investigated in a rat tail model, containing arteries and nerves structurally similar to those in the human hand. Tails were selectively vibrated for 1 or 9 days with the remainder of the animal at rest. One vibration bout of 4 h/day, 60 HZ, 5 g (49 m/s(2)) acceleration, injured endothelial cells. Injury was signaled by elevated immunostaining for NFATc3 transcription factor. Electron microscopy revealed that vibration for 9 days produced loss and thinning of endothelial cells, with activated platelets coating the exposed subendothelial tissue. Endothelial cells and arterial smooth muscle cells contained double membrane-limited, swollen processes indicative of vasoconstriction-induced damage. Laser doppler surface recording demonstrated that 5 min of vibration significantly diminished tissue blood perfusion. These findings indicate that early injury involves vasoconstriction and denuding of the arterial endothelium. PMID- 11932971 TI - The natural history of long thoracic and spinal accessory neuropathies. AB - A cohort of 106 patients with electrodiagnostically confirmed long thoracic neuropathy (50 patients) or spinal accessory neuropathy (56 patients) seen at the Mayo Clinic over a 22-year period were retrospectively studied to better understand the natural history of these disorders and to determine the role of electrodiagnostic testing in predicting prognosis. Mean follow-up was 48 and 50 months, respectively. Good functional recovery was generally observed regardless of the results of electrodiagnostic studies, but improvement in the amplitude of the spinal accessory compound muscle action potential on serial nerve conduction studies tended to predict a good outcome. No electrodiagnostic findings correlated with poor outcome. Traumatic neuropathies generally did worse than neuropathies of other causes. In spinal accessory neuropathies, involvement of the dominant limb, scapular winging, and impaired arm elevation were associated with a poor outcome. Our data suggest that, contrary to other focal neuropathies, the electrodiagnostic findings do not predict functional outcome in these neuropathies. PMID- 11932972 TI - Facioscapulohumeral muscular dystrophy with EcoRI/BlnI fragment size of more than 32 kb. AB - Facioscapulohumeral muscular dystrophy (FSHD) is associated with the deletion of a variable number of 3.3-kb subunits of a tandemly arranged repeat (D4Z4) on chromosome 4q35. EcoRI/BlnI fragments in the range of 10-35 kb are currently defined as disease-associated. Diagnosis of FSHD is frequently complicated by interchromosomal exchange with a homologous locus on 10q26. We present clinical and laboratory data of six subjects from two unrelated families with a marked FSHD phenotype and EcoRI/BlnI fragments of 39 and 33 kb, respectively. Origin on chromosome 4q35 was confirmed by haplotype analysis in the first family and was supported by pulsed field gel electrophoresis data in the second family. Our data further confirm the existence of a region of overlap of normal and pathological fragments. Fragments from this region can obviously be associated with marked FSHD phenotypes. Furthermore, application of linked markers and resolution of all EcoRI/BlnI fragments by pulsed field gel electrophoresis in addition to routine laboratory tests considerably augments the information obtained from molecular tests, upon which genetic counselling can then be based. PMID- 11932973 TI - Detection and prevalence of alpha-latrotoxin-like effects of serum from patients with Guillain-Barre syndrome. AB - Anti-GQ1b antibodies are associated with the Miller Fisher syndrome (MFS), a variant of the Guillain-Barre syndrome (GBS). In the ex vivo mouse diaphragm, anti-GQ1b-positive MFS serum induces muscle fiber twitching, a temporary dramatic increase of spontaneous quantal acetylcholine release, and transmission blockade at neuromuscular junctions (NMJs). These effects resemble those of alpha latrotoxin (alpha-LTx) and are induced by antibody-mediated activation of complement. We developed an assay for detection of the alpha-LTx-like effect, using muscle fiber twitching as indicator. We tested 89 serum samples from GBS, MFS, and control subjects, and studied correlations with clinical signs, anti ganglioside antibodies, micro-electrode physiology, and complement deposition at NMJs. Twitching was observed with 76% of the MFS and 10% of the GBS samples. It was associated with ophthalmoplegia and anti-GQ1b antibodies in patients, and with increased spontaneous acetylcholine release and C3c-deposition at mouse NMJs. This study strongly suggests that antibodies to GQ1b (with cross-reactivity to related gangliosides) are responsible for the alpha-LTx-like activity. The twitching assay is an efficient test for detection of this effect, and allows for screening of large numbers of samples and modifying drugs. PMID- 11932974 TI - Effects of training on contractile properties of paralyzed quadriceps muscle. AB - Effects of two different training regimens on the contractile properties of the quadriceps muscle were studied in six individuals with spinal cord injury. Each subject had both limbs trained with the two regimens, consisting of stimulation with low frequencies (LF) at 10 HZ or high frequencies (HF) at 50 HZ; one limb of each subject was stimulated with the LF protocol and the other with the HF regimen. Twelve weeks of daily training increased tetanic tension by approximately 20%, which was not significantly different between training regimens. Interestingly, after HF but not LF training, the unusual high forces at the low frequency range of the force-frequency relationship decreased, possibly due to a reduced activation per impulse. After LF but not HF training, force oscillation amplitudes declined (by 33%) as relaxation tended to slow, which may have opposed possible effects of reduced activation as seen after HF training. Finally, fatigue resistance also increased rapidly after LF training (by 43%) but not after HF training. These results indicate that different types of training may selectively change different aspects of function in disused muscles. PMID- 11932975 TI - Delay in initiation and termination of muscle contraction, motor impairment, and physical disability in upper limb hemiparesis. AB - The purpose of this study was to describe the relationship between the delay in initiation and termination of muscle contraction and clinical measures of motor impairment and physical disability in the affected upper limb of patients with hemiparesis. Electromyographic (EMG) activity of 26 long-term survivors of stroke was recorded during isometric wrist flexion and extension. Upper limb motor impairment and disability were assessed with the Fugl-Meyer motor assessment (FMA) and arm motor ability test (AMAT), respectively. Delay in initiation and termination of muscle contraction was significantly prolonged in the paretic arm. However, the delay was not significantly affected by stroke type, stroke level, side of hemiparesis, or presence of aphasia. Delay in initiation and termination of muscle contraction correlated significantly with FMA and AMAT. Abnormally delayed initiation and termination of muscle contraction may contribute to hemiparetic upper limb motor impairment and physical disability in hemiparetic patients. PMID- 11932976 TI - Effects of experimental muscle pain on electromyographic activity of masticatory muscles in the rat. AB - The aim of the present study was to investigate the effects of noxious chemical stimulation of a jaw muscle on postural electromyographic (EMG) activity from several masticatory muscles in lightly anesthetized rats. Unilateral injection of a substance known to induce acute muscle pain (5% NaCl) or longer duration of pain with inflammation (mustard oil) was made into the masseter muscle. The changes in EMG activity following the injection were recorded from the injected and contralateral masseter muscles and the ipsilateral digastric muscle. The algesic chemicals produced a significant but transient increase in EMG activity in all three muscles. The data from the present study and similar observations from clinical and experimental human studies suggest that increased activity from muscle nociceptors is not sufficient to produce a prolonged increase in postural EMG activity. Therefore, the development and maintenance of chronic jaw muscle pain does not appear to result from a feedback cycle mechanism. PMID- 11932977 TI - Pathophysiology of weakness in a patient with congenital end-plate acetylcholinesterase deficiency. AB - A Japanese patient with congenital end-plate acetylcholinesterase (AChE) deficiency developed severe proximal and truncal muscle weakness with preservation of distal strength. Electrophysiological studies included a train of stimuli at 3 HZ, which induced a marked decremental response in the deltoid but not in the first dorsal interosseous (FDI) muscle. Single fiber electromyography (EMG) revealed a high blocking rate (23.1 +/- 30.5%, n = 13) with a markedly increased jitter (mean consecutive difference [MCD] 297 +/- 218 micros) in the deltoid, but a low blocking rate (6.2 +/- 7.4%, n = 16) despite an equally increased jitter (MCD 227 +/- 147 micros) in the FDI. In vitro microelectrode study and computer simulation suggested that the combination of a large jitter and a low blocking rate may be ascribed to a reduced end-plate potential (EPP) amplitude with an abnormally prolonged decay time constant (tau). These characteristics may constitute the primary underlying pathophysiologic mechanism in our patient and in similar cases of congenital myasthenic syndrome. PMID- 11932978 TI - Application of NMR spectroscopy to monitoring MELAS treatment: a case report. AB - 1H magnetic resonance spectroscopy (MRS) of the brain and (31)P MRS and saturation transfer of resting skeletal muscle were used to investigate intracellular metabolites and fluxes through the creatine kinase (CK) reaction in a patient with the syndrome of mitochondrial myopathy, encephalopathy, lactic acidosis, and strokelike episodes (MELAS). Acute cortical lesions were characterized by severely elevated lactate levels and reduced concentrations of N acetylaspartyl compounds, glutamate, and myo-inositol. Similar but less extreme alterations were also observed in gray matter regions that appeared normal on magnetic resonance images. Investigation of the gastrocnemius muscle at rest demonstrated a reduced phosphocreatine level, elevated concentrations of inorganic phosphate and free adenosine 5'-diphosphate, and an abnormally low phosphorylation potential. Besides a moderately increased muscular phosphocreatine concentration, none of the metabolic disturbances detected on MRS improved with oral creatine supplementation. Forward and reverse fluxes through the CK reaction did not significantly change upon creatine treatment. Follow-up MRS investigations may thus provide objective markers of treatment response in vivo without the hazards or inconvenience of biopsy. PMID- 11932980 TI - Obturator neuropathy: causes and outcome. AB - To study causes of obturator neuropathy and to correlate them with outcome, we retrospectively studied patients seen at the Mayo Clinic electromyography (EMG) laboratory from 1975 through 1999 with a diagnosis of obturator neuropathy. Twenty-two patients with obturator neuropathy were identified. The clinical outcome of patients with acute obturator neuropathy treated conservatively was good regardless of etiology or severity. PMID- 11932979 TI - Parallel expression of neurotrophic factors and their receptors in chronic inflammatory demyelinating polyneuropathy. AB - The mRNA levels of nerve growth factor (NGF), glial cell line-derived neurotrophic factor (GDNF), ciliary neurotrophic factor (CNTF), leukemia inhibitory factor (LIF), and interleukin-6 (IL-6) were examined in sural nerves of 22 patients with chronic inflammatory demyelinating polyneuropathy (CIDP). The mRNAs for NGF, GDNF, LIF, and IL-6 were upregulated, whereas CNTF mRNA was downregulated significantly in the nerves. The NGF, GDNF, and CNTF, but not LIF mRNA expressions were parallel to those of the cognate receptors, suggesting that these cognate soluble receptors effectively present these factors to maintain and regenerate the axons. Furthermore, IL-6 mRNA expression was significantly parallel to both binding and signal-transducing receptor expression, implying a role of the IL-6 signal for non-neuronal cells in CIDP. These findings indicate that multiple neurotrophic growth factors and cytokines are expressed cooperatively with their concomitant receptors in the nerve lesions of CIDP and play an important role particularly in nerve repair. PMID- 11932981 TI - Electrophysiological evaluation of conduction in the most proximal motor root segment. AB - Root conduction time (RCT), defined as the time difference between M-wave latency by cervical magnetic stimulation (CMS) and the total peripheral motor conduction time calculated from the shortest F-wave latency, was investigated in patients with inflammatory demyelinating neuropathies (IDP) and amyotrophic lateral sclerosis (ALS). The minimal threshold for CMS also was studied. In the IDP patients, conduction in the proximal motor root segment was considered abnormal in 52% by the RCT and in 47% by the minimal threshold for CMS, whereas both were normal in 85% of the ALS patients. These findings suggest that the RCT and minimal threshold for CMS might be additional parameters for evaluating motor nerve conduction in IDP. PMID- 11932982 TI - Receptors for NGF and GDNF are highly expressed in human peripheral nerve neuroma. AB - Immunhistochemistry of 10 traumatic human peripheral nerve neuromas and 5 normal digital nerves revealed a high immunoreactivity for the nerve growth factor (NGF) receptors p75 and trkA and the glial cell line-derived neurotrophic factor (GDNF) receptor GFRalpha-1. Semiquantitative image analysis showed a significantly increased trkA immunoreactivity in the neuroma group. The presence of the receptors may provide a way to influence therapy of peripheral nerve neuroma by administration of neurotrophins or other substances with binding sites similar to those of a neurotrophic factor. PMID- 11932983 TI - Myotonic dystrophy type 1 (DM1) presenting with laryngeal stridor and vocal fold paresis. AB - Myotonic dystrophy type 1 (DM1) is the most common inherited muscle disorder and may present in numerous ways due to characteristic multisystem involvement. We report a 47-year-old man who presented with an 8-year history of slowly progressive dyspnea and episodic stridor. The laryngeal paresis was documented with videostroboscopy and laryngeal electromyography, and treated with tracheostomy and antimyotonia agents. PMID- 11932984 TI - Idiopathic bilateral diaphragmatic paralysis. AB - A 41-year-old man complained of subacute onset of dyspnea and pain in the neck and chest. He was diagnosed with bilateral diaphragmatic paralysis, based on clinical inspection of the breathing pattern and transdiaphragmatic pressure recording, and was trained to use a portable bi-level positive airway pressure apparatus (BiPAP). Needle electromyography showed profuse fibrillation potentials and positive waves in the diaphragm, more abundant on the right than left side, and no response to phrenic nerve stimulation. Other muscles were not involved. Follow-up examinations, performed at 9 and 12 months after onset of paralysis, demonstrated a slow but progressive improvement of the patient's respiratory function, together with the appearance of reinnervation potentials in the diaphragm, and polyphasic, long-latency responses to phrenic nerve stimulation. The subacute onset of the paralysis associated with local pain, and its subsequent recovery, suggest bilateral proximal lesions in the phrenic nerves. In the absence of traumatic or metabolic causes, these findings suggest that the phrenic nerve can be a target in idiopathic neuritis. PMID- 11932986 TI - Teratogenesis of holoprosencephaly. AB - Teratogenic causes of holoprosencephaly are critically assessed. A brief general review of holoprosencephaly is followed by four tables summarizing etiologic factors. Subjects evaluated here include: 1) maternal diabetes; 2) ethyl alcohol; 3) retinoic acid; 4) mutated genes and teratogens involving the sonic hedgehog signaling network and cholesterol biosynthesis; and 5) cholesterol trafficking, sterol adducts, target tissue response, and sterol sensing domain. PMID- 11932988 TI - High risk of malignancy in mosaic variegated aneuploidy syndrome. AB - Fourteen cases of mosaic variegated aneuploidy (MVA) syndrome have been reported in the last 10 years. The phenotype of this rare condition has been quite consistent: severe microcephaly, growth deficiency, mild physical anomalies, and mental retardation. We describe here a young boy in whom MVA syndrome is associated to myelodysplasia with a monosomy 7 bone marrow clone. At the age of 3 years, myelodysplasia progressed to an acute lymphoblastic leukemia, and the patient died soon after. Several syndromes with short stature and severe microcephaly, such as the Seckel and Nijmegen syndromes, comprise hematological findings and chromosome instability. However, chromosome instability was not confirmed in our patient. MVA with hematological findings has not been reported before, but 3 patients of 14 (21%) have developed a malignancy (rhabdomyosarcoma, acute lymphoblastic leukemia, and nephroblastoma). Therefore, we propose that MVA is a condition predisposing to neoplasia. PMID- 11932989 TI - Hereditary intraosseous vascular malformation of the craniofacial region: an apparently novel disorder. AB - Primary intraosseous vascular anomaly, previously called intraosseous hemangioma, is a very rare malformation that is usually seen in the vertebral column and in the skull. It is exclusively described in sporadic cases and no hereditary component has yet been reported. The most commonly affected bones in the skull are the mandible and the maxilla, and life-threatening bleeding after a simple tooth extraction is frequently observed. Here, we report two consanguineous families containing a total of four affected patients manifesting primary intraosseous vascular malformation (VMOS (vascular malformation osseous)) of the craniofacial region. The phenotypic expression is remarkably similar in both families. The characteristic findings include severe blood vessel expansions within the craniofacial bones and midline abnormalities such as diastasis recti, supraumbilical raphe, and hiatus hernia. Malformation is restricted to the mandibular and maxillary area in the prepubertal age, and rapid expansion starts after age 12 or 13. A 15-year follow-up of one of the patients demonstrated that the vascular malformation did not extend beyond the craniofacial region despite severe involvement of almost all bones in the skull. Detailed clinical and radiological evaluation provided neither evidence of soft-tissue involvement nor any sign of gross arterial, venous, or combined malformations, indicating that bone changes are a primary rather than a secondary effect due to any other vascular anomaly in the craniofacial region. An antibody against a universal proliferation marker, Ki-67, detected nonproliferative, single-layered endothelial cells, suggesting that this abnormality is a vascular malformation rather than a hemangioma. alpha-actin staining (antibody against perivascular tissue such as smooth muscle cells (SMCs) and/or pericytes) demonstrated that pathologic vessels lost their surrounding supportive tissues, as was previously seen in other types of vascular anomaly. Homozygosity mapping excluded the following loci and/or genes: multiple cutaneous venous malformation (VMCM1; gene, TIE2) on chromosome 9p21; venous malformation with glomus cells (VMGLOM) on chromosome 1p22-p21; hereditary hemorrhagic telangiectasia type 1 (HHT1; gene, endoglin) and type 2 (HHT2; gene, activin) on chromosomes 9q34.1 and 12q11-q14, respectively; and cerebral cavernous malformation type 1 (CCM1; gene, KRIT1), type 2 (CCM2), and type 3 (CCM3) on chromosomes 7q11.2-q21, 7p15-p13, and 3q35.2 q27, respectively. To the best of our knowledge, this is a new disorder, which we call hereditary intraosseous vascular malformation of the craniofacial region. PMID- 11932990 TI - Autism and the X chromosome: no linkage to microsatellite loci detected using the affected sibling pair method. AB - The etiology of autism spectrum disorders (ASDs) is poorly understood, although it is clear that genetic factors play a major role. ASDs appear to be a heterogeneous group of disorders, making genetic analysis difficult in the absence of etiologically definable subgroups. The excess of males in the affected population has led to suggestions that an X-linked locus could play a role in the causation of autism or a related pervasive developmental disorder. To examine this, we have investigated the genotypes of 31 families with two or more affected boys, at a series of 16 highly polymorphic loci distributed along the X chromosome with an average interlocus distance of 12 cM, in order to identify regions of significantly increased concordance among pairs of affected brothers. No locus tested showed a significant increase in concordance, supporting findings by others that there are no genes of major effect located on the X chromosome that contribute to increased susceptibility to ASD. PMID- 11932992 TI - Duodeno-jejunal atresia with volvulus, absent dorsal mesentery, and absent superior mesenteric artery: a hereditary compound structure in duodenal atresia? AB - Duodenal atresia predominantly represents an isolated entity, but could also be part of a complexly structured intestinal malformation. We report four children, including two from one family, with duodeno-jejunal atresia associated with malrotation, volvulus, and absent parietal attachment of the mesentery. Gross absence of the mesentery and absence of distal parts of the superior mesenteric artery were the most remarkable findings. The small intestine was supplied retrogradely from the right colic artery. Consideration of embryological theories of this malformation takes into account the important role of the duodeno-jejunal flexure during the process of entry of the small bowel loops into the abdomen. Familial occurrence suggests autosomal recessive inheritance. We propose differentiating the pathoanatomical findings in our patients from classical apple peel small bowel syndrome (APSB). PMID- 11932991 TI - Niemann-Pick disease type B: an unusual clinical presentation with multiple vertebral fractures. AB - We report here a unique case of a 55-year-old woman presenting with a clinical picture of Parkinson disease, severe back pain, splenomegaly, and pronounced dyspnea. Radiographic examination of the spine showed multiple vertebral fractures. Niemann-Pick disease type B was diagnosed by findings of lipid-loaded histiocytes and a strongly reduced sphingomyelinase enzyme activity. She was homozygous for the deletion of codon 608 (delR608), which encodes an arginine residue in the Acid Sphingomyelinase gene. To investigate the cause of the unusual vertebral fractures, we screened for polymorphisms previously described as possibly associated with increased risk for osteoporosis and fractures. Our patient was heterozygous for the polymorphisms of the vitamin D receptor gene, the estrogen receptor gene, and the collagen 1A1gene. Increased physical activity after Parkinson treatment, a genetic predisposition, together with worsening disease due to interfering medications could explain the dramatic presentation of this patient. She was treated with cholesterol lowering drugs such as statins to decrease sphingomyelin synthesis, avoidance of drugs that inhibit sphingomyelinase, and bisphosphonates. No new fractures have occurred, but the interstitial lung disease has progressed. PMID- 11932993 TI - A Thai mother and son with distal symphalangism, hypoplastic carpal bones, microdontia, dental pulp stones, and narrowing of the zygomatic arch: a new distal symphalangism syndrome? AB - A Thai mother and son with distal symphalangism and other associated abnormalities are reported. Distal and middle phalanges of fingers and toes 2-5 were either aplastic/hypoplastic or fused between the corresponding digits. The second fingers and fourth fingernails were most severely affected in both patients. The mother's hands were less severely affected; the middle and distal phalanges of her hands were malformed and fused. Besides the absence of fusion lines, the shape of the fused middle and distal phalanges was quite different from that of other types of fusion, i.e., fused bones in both patients did not maintain the normal configuration of bone, referring to as "middle-distal phalangeal complex". Distal symphalangism was observed in toes 2-5 of the mother and in toe 3 of the son. Both patients had additional clinical manifestations such as narrowing of the zygomatic arch, dental pulp stone, microdontia of a mandibular permanent central incisor, cone-shaped epiphyses of middle phalanges of fingers, and absence of scaphoid, trapezium, trapezoid, and pisiform bones. Mutation analysis of NOG and ROR2, the genes responsible for proximal symphalangism and brachydactyly type B, respectively, was negative. PMID- 11932994 TI - Sister and brother with Vici syndrome: agenesis of the corpus callosum, albinism, and recurrent infections. AB - A sister and brother with Vici syndrome are described. They both had oculocutaneous albinism, agenesis of the corpus callosum, cataracts, and cardiomyopathy. They were born to healthy unrelated parents, and had postnatal growth retardation, profound developmental delay, hypotonia, and cataracts. The sister had recurrent infections, and died of progressive heart failure at age 19 months. The brother is alive at age six months with mild cardiomyopathy, and had a single episode of acute bronchitis at age three months. Review of the clinical manifestations of the sibs we described and six children reported in the literature indicates that Vici syndrome is a distinct clinical entity. Its main clinical manifestations include growth retardation, profound developmental delay, hypotonia, albinism, agenesis of the corpus callosum, cataracts, cardiomyopathy, and recurrent infections. The occurrence of the syndrome in three pairs of sibs of both sexes born to unaffected parents supports autosomal recessive inheritance. PMID- 11932995 TI - De novo translocation t(5;6)(q35;q21) in an infant with Walker-Warburg syndrome. PMID- 11932996 TI - What is the birth defect risk associated with consanguineous marriages? PMID- 11932997 TI - Premature thelarche in Rubinstein-Taybi syndrome. PMID- 11932998 TI - Two-hit model for tumorigenesis of nevoid basal cell carcinoma (Gorlin) syndrome associated hepatic mesenchymal tumor. PMID- 11932999 TI - Diagnosis of Ambras syndrome: comments on complex cytogenetic rearrangement of chromosome 8q in a case of Ambras syndrome. PMID- 11933000 TI - Folic acid supplementation and neural tube defects. PMID- 11933002 TI - Health disparities research: exploring different research pathways. PMID- 11933003 TI - Life values before versus after a breast cancer diagnosis. AB - The main aim of this study was to investigate whether women's life values change with a breast cancer diagnosis. In addition, associations between life values and anxiety/depression ratings were investigated. Life value changes were prospectively studied in 517 women recalled for further examination after attending mammographic screening, 38 of whom were diagnosed with primary breast cancer. Life values were assessed by a study-specific version of a life value questionnaire, including ratings of the perceived attainment and importance of seven life value dimensions. Three months after being recalled, women diagnosed with primary breast cancer reported a reduction of the attainment and the importance of Health. In addition, these women reported changes in the perceived importance of Responsibility and Involvement. High levels of anxiety and depression in particular were associated with large discrepancies between attainment and importance for some life values. This suggests that changes in the perceived importance of some life values may constitute one part of women's psychological adaptation to a breast cancer diagnosis. PMID- 11933004 TI - Prevalence, prevention, and treatment of pressure ulcers: descriptive study in 89 institutions in the Netherlands. AB - The purpose of the present study was to assess the prevalence of pressure ulcers and the use of Dutch guidelines for the prevention and treatment of pressure ulcers. A survey of 16,344 patients in 89 health care institutions on 1 day showed a mean prevalence of pressure ulcers of 23.1%. It was found that Dutch guidelines on some aspects of prevention and treatment of pressure ulcers were not being followed. Only 53% of the patients who should have been positioned on a support surface were positioned on such a device. Fewer than one-third of the patients who should have been repositioned, should have received nutritional support, or should have been educated received these interventions, and only 33.6% of all pressure ulcers were dressed as recommended. More attention to the dissemination and implementation of the guidelines is needed to reduce this high prevalence of pressure ulcers. PMID- 11933005 TI - A conceptual framework for understanding the process of family caregiving to frail elders in Taiwan. AB - This descriptive, correlational study of family caregivers (N=125) tested a conceptual framework for family caregiving to frail elders in Taiwan, using the concept of "finding a balance point" derived from a previous qualitative study. It was hypothesized that caregivers who were better able to find a balance point among competing needs would provide better-quality care to frail elders, which would lead to more positive caregiver and family outcomes. After controlling for the influence of caregiving characteristics and caregiving factors, finding a balance point significantly explained 7% of the variance in overall caregiving consequences. The findings of this study add a new perspective to the caregiving process in Taiwan. PMID- 11933006 TI - Modeling condom-use stage of change in low-income, single, urban women. AB - This study was undertaken to identify and test a model of the cognitive antecedents to condom use stage of change in low-income, single, urban women. A convenience sample of 537 women (M=30 years old) attending two urban primary health care settings in western New York State anonymously completed questionnaires based primarily on two leading social-cognitive models, the transtheoretical model and the information-motivation-behavioral skills model. We used structural equation modeling to examine the direct and indirect effects of HIV-related knowledge, social norms of discussing HIV risk and prevention, familiarity with HIV-infected persons, general readiness to change sexual behaviors, perceived vulnerability to HIV, and pros and cons of condom use on condom-use stage of change. The results indicated two models that differ by partner type. Condom-use stage of change in women with steady main partners was influenced most by social norms and the pros of condom use. Condom-use stage of change in women with "other" types (multiple, casual, or new) of sexual partners was influenced by HIV-related knowledge, general readiness to change sexual behaviors, and the pros of condom use. These findings suggest implications for developing gender-relevant HIV-prevention interventions. PMID- 11933007 TI - Trichotillomania: women's reports of treatment efficacy. AB - Trichotillomania (TTM) is an enigmatic disease characterized by an overwhelming compulsion to pull out one's hair, most notably scalp hair, but also eyebrows, eyelashes, and hair from the extremity, axillary, and pubic areas. Current estimates are that clinically significant hair pulling is manifested by 3.4% of all women during their lifetimes. The overall purpose of our study was to examine the efficacy of a wide variety of treatments as perceived by the women (N=44) who received them. The data were subjected to content analysis and are presented in a quantified manner. Common forms of treatments, including pharmacotherapy, psychotherapy, and behavior modification, were judged by the respondents to be relatively ineffective. Two forms of treatment were found to be effective by a high percentage of the women who employed them: Internet TTM groups and TTM groups associated with a national center. It is important for health care providers to be aware of these treatments, which have not been previously discussed in the literature. PMID- 11933009 TI - Sociospatial knowledge networks: appraising community as place. AB - This article introduces a new theory of geographical analysis, sociospatial knowledge networks, for examining and understanding the spatial aspects of health knowledge (i.e., exactly where health beliefs and knowledge coincide with other support in the community). We present an overview of the theory of sociospatial knowledge networks and an example of how it is being used to guide an ongoing ethnographic study of health beliefs, knowledge, and knowledge networks in a rural community of African Americans, Latinos, and European Americans at high risk for, but not diagnosed with, type 2 diabetes mellitus. We believe that the geographical approach to understanding health beliefs and knowledge and how people acquire health information presented here is one that could serve other communities and community health practitioners working to improve chronic disease outcomes in diverse local environments. PMID- 11933008 TI - Measuring health beliefs in Spanish-speaking Mexican Americans with type 2 diabetes: adapting an existing instrument. AB - Mexican Americans bear a disproportionate burden of type 2 diabetes, and although clinicians and researchers are beginning to develop culturally competent interventions, few instruments are available for measuring variables relevant to health outcomes. The purpose of this study was to determine the psychometric properties of a diabetes-related health belief instrument translated for use with Spanish-speaking Mexican Americans. Three hundred and twenty-six Spanish-speaking Mexican Americans from Starr County, a Texas-Mexico border community, diagnosed with type 2 diabetes participated in the study. Factor analysis was used to examine the factor structure of the instrument, and reliability testing was conducted to determine the internal consistency of the resulting factors. The final instrument had 25 items with four subscales-Barriers, Social Support for Diet, Impact of Job on Therapy, and Benefits of Therapy-and one item to measure, Control. Internal consistency coefficients for these subscales were .56, .62, .86, and .90, respectively. The translated instrument needs to be tested over time with other Hispanic populations. PMID- 11933010 TI - Flow cytometry used for the analysis of calcium signaling induced by antigen specific T-cell activation. AB - BACKGROUND: In this study, the effect of antigen-presenting cells (APC), peptide concentration, and CD28 costimulation on calcium signaling, induced by antigen specific T-cell activation, was studied by flow cytometry. METHODS: We used two experimental approaches, which differed in their time scale and in the duration of the T cell-APC interaction, to measure the increase of intracellular free calcium levels ([Ca(2+)](i)) in activated T cells: (1) Fluo-3-loaded T cells were activated by cocentrifugation with peptide-loaded APC and the kinetics of fluorescence intensity changes was monitored continuously and (2) peptide-loaded APC and T cells were mixed, cocultured, and the fluorescence intensity was measured at various time intervals. RESULTS: The calcium signal of T cells was dependent on the APC as demonstrated by the ratio of cells exhibiting high versus low fluorescence intensity and by the magnitude of the calcium signal in the activated population. Short-term interaction of T cells with less potent APC or with efficient APC in the presence of low antigen concentration resulted in decreased calcium signaling. CD28-mediated costimulation enhanced the magnitude and sustained the increase of intracellular calcium levels. In line with the strong and sustained calcium signals, the activation of the calcium-dependent transcription factors NF-AT, AP-1, and NF-kappaB was induced. CONCLUSIONS: Flow cytometric methods, feasible for the rapid and flexible analysis of calcium signaling upon antigen-specific T-cell activation, were established. Kinetics of the increase of mean fluorescence intensity reflected the calcium response of the total cell population whereas statistical analysis of fluorescence intensity at selected time points provided information on the activation state of single cells. PMID- 11933011 TI - Spatial distribution patterns of interphase centromeres during retinoic acid induced differentiation of promyelocytic leukemia cells. AB - BACKGROUND: The pericentromeric heterochromatin is an important element for the regulation of gene silencing. Its spatial distribution during interphase appears to be cell-type specific. This study analyzes three-dimensional (3D) centromere distribution patterns during cellular differentiation along the neutrophil pathway. METHODS: Differentiation of the promyelocytic leukemia cell line NB4 was induced by retinoic acid. Centromeres in interphase nuclei were visualized by immunofluorescence staining of centromere-associated proteins with CREST serum. 3D images of nuclei were obtained by confocal microscopy. Automated methods for the segmentation of point-like objects in 3D images were implemented to detect the position of centromeres. Features of centromere localization patterns were determined by constructing the minimal spanning tree of the centromere distribution. RESULTS: In differentiated NB4 cells, the number of centromere conglomerates (chromocenters) was decreased and the distance between chromocenters was increased as compared with untreated controls. The nuclear volume did not differ between the two groups. CONCLUSIONS: The measured rearrangement of centromeres indicates a progressive clustering of heterochromatin and a global remodeling of interphase chromosome territories during differentiation of NB4 cells. The developed methods for the analysis of 3D centromere distribution patterns provide the opportunity for a fast and objective analysis of heterochromatin remodeling. PMID- 11933012 TI - Discrimination of DNA and RNA in cells by a vital fluorescent probe: lifetime imaging of SYTO13 in healthy and apoptotic cells. AB - BACKGROUND: Of the few vital DNA and RNA probes, the SYTO dyes are the most specific for nucleic acids. However, they show no spectral contrast upon DNA or RNA binding. We show that fluorescence lifetime imaging using two-photon excitation of SYTO13 allows differential and simultaneous imaging of DNA and RNA in living cells, as well as sequential and repetitive assessment of staining patterns. METHODS: Two-photon imaging of SYTO13 is combined with lifetime contrast, using time-gated detection. We focus on distinguishing DNA and RNA in healthy and apoptotic Chinese hamster ovary cells. RESULTS: In healthy cells, SYTO13 has a fluorescence lifetime of 3.4 +/- 0.2 ns when associated with nuclear DNA. Bound to RNA, its lifetime is 4.1 +/- 0.1 ns. After induction of apoptosis, clusters of SYTO13 with fluorescence lifetime of 3.4 +/- 0.2 ns become apparent in the cytoplasm. They are identified as mitochondrial DNA on the basis of colocalization experiments with the DNA-specific dye, DRAQ5, and the mitochondrial-specific dye, CMXRos. Upon progression of apoptosis, the lifetime of SYTO13 attached to DNA shortens significantly, which is indicative of changes in the molecular environment of the dye. CONCLUSIONS: We have characterized SYTO13 as a vital lifetime probe, allowing repetitive and differential imaging of DNA and RNA. PMID- 11933013 TI - Mitochondrial and nonmitochondrial reduction of MTT: interaction of MTT with TMRE, JC-1, and NAO mitochondrial fluorescent probes. AB - BACKGROUND: Bioreduction of water-soluble tetrazolium salts (e.g., MTS, XTT, and MTT) to their respective formazans is generally regarded as an indicator of cell "redox activity." The reaction is attributed mainly to mitochondrial enzymes and electron carriers. However, MTT reduction may also be catalyzed by a number of other nonmitochondrial enzymes. The goal of this work was to establish the sites of MTT reduction in intact HepG2 human hepatoma cells in culture. METHODS: In order to establish the subcellular localization of the sites of reduction of MTT, we imaged the formation of MTT-formazan deposits using backscattered light confocal microscopy. Mitochondria were visualized in viable cells using fluorescent dyes that bind in a manner dependent (JC-1 and TMRE) or independent (NAO) of mitochondrial electric potential. RESULTS: Only 25-45% of MTT-formazan was associated with mitochondria after 25 min of incubation. No more than 25% of the mitochondrial area on images was occupied by MTT-formazan. Mitochondrial fluorescence of TMRE, NAO, and the monomeric form of JC-1 decreased rapidly in cells incubated with MTT. However, the intensity of fluorescence of JC-1 aggregates dropped by less than 30% at the onset of incubation and remained constant as reduction of MTT proceeded further. CONCLUSIONS: (1) Most of MTT formazan deposits are not coincident with mitochondria. (2) Monomeric JC-1, as well as TMRE and NAO, accumulating in mitochondria may be displaced by MTT. Thus, the presence of positively charged organic compounds (like MTT) may distort measurements of mitochondrial transmembrane electric potential, which are based on accumulation of fluorescent dyes. PMID- 11933014 TI - Simultaneous detection of bacteria expressing GFP and DsRed genes with a flow cytometer. AB - BACKGROUND: In this study, Escherichia coli cells producing red fluorescent protein of Discosoma sp. (drFP583 DsRed) were investigated with flow cytometry by using 488 nm excitation. We also studied whether green fluorescent protein (GFP) and drFP583 could be detected simultaneously from a single bacterial cell. METHODS: Plasmids pDsRed and pEGFP were used for the production of drFP583 and enhanced GFP, respectively, in E. coli MC1061 cells. To produce enhanced GFP and drFP583 simultaneously, plasmids pG9R and pG19R were constructed. These encode tandem fusions of enhanced GFP and drFP583 to ensure similar production levels for both proteins. RESULTS: Bacteria producing enhanced GFP and drFP583 were found to be brightly green and red fluorescent, respectively. Production of enhanced GFP and drFP583 fusion proteins resulted in bacteria that emitted both green and red fluorescence, which was detected easily by a flow cytometer using single laser excitation. Previously reported tetramerization of drFP583 did not restrict its use as a reporter gene, although it maturated significantly slower than enhanced GFP. CONCLUSIONS: The results show that enhanced GFP and drFP583 proteins can be detected simultaneously from single bacteria with a standard flow cytometer with simple optical configuration. PMID- 11933015 TI - Imaging technique implemented in CellTracks system. AB - BACKGROUND: We developed the CellTracks cell analysis system that, similar to flow cytometry, yields multiparameter information by which the cells can be differentiated. We describe the implementation of a laser scanning imaging method in the system. Image analysis of the cells improves the specificity of cell classification, especially in cases where the particular cells are found relatively infrequently and one has to discriminate between artifacts and real events. METHODS: Fluorescent images of immunomagnetically labeled and aligned cells are obtained by passing the cells through a laser focus. The laser focus is smaller than the objects and subsequent frames captured by a regular surveillance CCD camera with a frame grabber board represent different parts of the cells. Complete images of the cells are constructed by shifting each image with respect to each other and adding individual pixel values. RESULTS: The power of combining a fluorescent image with multiparametric data is demonstrated by imaging fluorescent and magnetically labeled beads and cells. The image gives additional information about the dye distribution across the objects. Changes in dye distribution as a function of time were observed in leukocytes labeled with the red fluorescent label, Oxazine750, which are imaged at different time intervals. CONCLUSIONS: An imaging technique implemented in the CellTracks system provides high-resolution fluorescent images of events previously identified by the system. The images of the fluorescent cells enhance the ability to classify rare events. PMID- 11933016 TI - Automated identification of diploid reference cells in cervical smears using image analysis. AB - BACKGROUND: Acquisition of DNA ploidy histograms by image analysis may yield important information regarding the behavior of premalignant cervical lesions. Accurate selection of nuclei for DNA measurement is an important prerequisite for obtaining reliable data. Traditionally, manual selection of nuclei of diagnostic and reference cells is performed by an experienced cytotechnologist. In the present study, a method for the fully automated identification of nuclei of diploid epithelial reference cells in Feulgen- restained Papanicolaou (PAP) smears is described. METHODS: The automated procedure consists of a decision tree implemented on the measurement device, containing nodes with feature threshold values and multivariate discriminant functions. Nodes were constructed to recognize debris and inflammatory cells, as well as diploid and nondiploid epithelial cells of the uterine cervix. Evaluation of the classifier was performed by comparing resulting diploid integrated optical densities with those from manually selected reference cells. RESULTS AND CONCLUSION: On average, automatically acquired values deviated 2.4% from manually acquired values, indicating that the method described in this paper may be useful in cytometric practice. PMID- 11933017 TI - A small-volume technique for simultaneous immunophenotyping and apoptosis detection in rat whole blood by four-color flow cytometry. AB - BACKGROUND: Cell permeabilization for the detection of intracellular molecules by flow cytometry is usually incompatible with whole blood. This article describes a new technique for the simultaneous detection of surface antigens and DNA content in rat whole blood. METHODS: In 20 microl of rat whole blood, DNA staining is obtained by permeabilization of cells using a standard red blood cell lysing reagent (Erythrolyse). Immunophenotyping and apoptosis detection by flow cytometry are achieved by using a combination of three surface markers (CD3, CD4, and CD8alpha) and a DNA binding dye (TO-PRO-3). RESULTS: After a 24-h incubation of whole blood with 1 microM dexamethasone, apoptotic lymphocytes were clearly distinguishable from normal lymphocytes by their reduced size and DNA content. The dexamethasone-induced percentage of apoptotic cells was 58.9 +/- 4.6 for CD4+ and 77.4 +/- 2.9 for CD8+ T cells, compared with 12.6 +/- 2.7 for CD4+ and 17.2 +/- 3.5 for CD8+ T cells in the absence of dexamethasone (data from 10 animals with duplicate samples). CONCLUSIONS: We have developed a new technique to permeabilize nucleated cells in microsamples of rat whole blood. The methodology allows simultaneous immunophenotyping and apoptosis detection in rat whole blood. PMID- 11933019 TI - Transferrin/transferrin receptor-mediated drug delivery. AB - Since transferrin was discovered more than half a century ago, a considerable effort has been made towards understanding tranferrin-mediated iron uptake. However, it was not until recently with the identification and characterization of several new genes related to iron homeostasis, such as the hemochromatosis protein HFE and the iron transporter DMT1, that our knowledge has been advanced dramatically. A major pathway for cellular iron uptake is through internalization of the complex of iron-bound transferrin and the transferrin receptor, which is negatively modulated by HFE, a protein related to hereditary hemochromatosis. Iron is released from transferrin as the result of the acidic pH in endosome and then is transported to the cytosol by DMT1. The iron is then utilized as a cofactor by heme and ribonucleotide reductase or stored in ferritin. Apart from iron, many other metal ions of therapeutic and diagnostic interests can also bind to transferrin at the iron sites and their transferrin complexes can be recognized by many cells. Therefore, transferrin has been thought as a "delivery system" for many beneficial and harmful metal ions into the cells. Transferrin has also be widely applied as a targeting ligand in the active targeting of anticancer agents, proteins, and genes to primary proliferating malignant cells that overexpress transferrin receptors. This is achieved by conjugation of transferrin with drugs, proteins, hybride systems with marcomolecules and as liposomal-coated systems. Conjugates of anticancer drugs with transferrin can significantly improve the selectivity and toxicity and overcome drug resistance, thereby leading to a better treatment. The coupling of DNA to transferrin via a polycation such as polylysine or via cationic liposomes can target and transfer of the extrogenous DNA particularly into proliferating cells through receptor mediated endocytosis. These kinds of non-viral vectors are potential alternatives to viral vectors for gene therapy, if the transfection efficiency can be improved. Moreover, transferrin receptors have shown potentials in delivery of therapeutic drugs or genes into the brain across blood-brain barrier. PMID- 11933020 TI - Chemical control of phospholipid distribution across bilayer membranes. AB - Most biological membranes possess an asymmetric transbilayer distribution of phospholipids. Endogenous enzymes expend energy to maintain the arrangement by promoting the rate of phospholipid translocation, or flip-flop. Researchers have discovered ways to modify this distribution through the use of chemicals. This review presents a critical analysis of the phospholipid asymmetry data in the literature followed by a brief overview of the maintenance and physiological consequences of phospholipid asymmetry, and finishes with a list of chemical ways to alter phospholipid distribution by enhancement of flip-flop. PMID- 11933021 TI - Nonsteroidal aromatase inhibitors: recent advances. AB - Aromatase is the cytochrome P450 enzyme responsible for the last step of estrogen biosynthesis, and aromatase inhibitors constitute an important class of drugs in clinical use for the treatment of breast cancer. Nonsteroidal aromatase inhibitors (NSAIs) are competitive inhibitors of aromatase, which bind to the enzyme active site by coordinating the iron atom present in the heme group of the P450 protein. Presently, third generation NSAIs are in use, and research efforts are being carried out both to identify new molecules of therapeutic interest and to clarify the mechanism of action. In this article, we present a survey of the compounds that have been recently reported as NSAIs, to provide a broad view on the general structure-activity relationships of the class. Moreover, starting from the current knowledge of the mechanistic aspects of aromatase action and from recent theoretical work on the molecular modeling of both enzyme and inhibitors, we try to indicate a way to integrate these different studies in view of a more general understanding of the aromeatase-inhibitor system. Finally, some aspects regarding the possible future development of the field are considered briefly. PMID- 11933022 TI - Bioactive taxoids from the Japanese yew Taxus cuspidata. AB - A series of new taxoids, named taxuspines A-H and J-Z (1-25) and taxezopidines A H and J-L (26-36), have been isolated together with 37 known taxoids (37-73) including paclitaxel (53) from the Japanese yew, Taxus cuspidata Sieb. et Zucc. (Taxaceae). These new taxoids possess various skeletons containing 5/7/6, 6/10/6, 6/5/5/6, 6/8/6, or 6/12-membered ring systems. Among the new taxoids, some non taxol-type compounds remarkably reduced CaCl(2)-induced depolymerization of microtubules, or increased cellular accumulation of vincristine in multidrug resistant tumor cells as potent as verapamil. On the other hand, chemical derivatization of taxinine (37), one of major taxoids obtained from this yew, led to the discovery of unusual reactions of taxinine derivatives. Here we describe our recent results on the isolation, structure elucidation, and bioactivity of these new and known taxoids and the formation of unexpected products of the unusual reactions of taxinine. PMID- 11933024 TI - High-performance liquid chromatographic profiling of fluorescent labelled N glycans on glycoproteins. AB - Monitoring protein glycosylation is becoming increasingly important as novel recombinant glycoprotein therapeutics, such as glycoprotein hormones, cytokines and clotting factors, are introduced into clinical use. In this report, we describe an HPLC strategy and an improved and simplified pre-column derivatization procedure to profile N-linked glycans obtained from a variety of commercially available glycoproteins as examples. N-Glycans were first released by peptide:N-glycosidase F and labelled with the fluorescent label, 4 aminobenzoic acid by reductive amination. The labelled N-Glycans were then resolved by normal-phase HPLC and the N-glycan profile could be further improved by separating the N-glycans first according to charge by anion-exchange HPLC prior to the normal-phase HPLC. If required, identification of the fractionated derivatized oligosaccharides can be determined by mass spectrometry. The whole profiling process is simple and can be implemented in most laboratories. Because of the high sensitivity, batch glycan-analysis of low-yield recombinant glycoproteins such as samples in ampoules or obtained in the early stage of production development is possible. PMID- 11933023 TI - Validation of a high-performance liquid chromatographic method for the enantiospecific quantitation of zopiclone in plasma. AB - Zopiclone is a hypnosedative with clinical effects similar to benzodiazepines but thought to have less potential for rebound insomnia and withdrawal effects. Zopiclone is administered as a racemic mixture, and an enantiospecific method of analysis of zopiclone in plasma is desirable in the study of pharmacokinetic drug interactions. We report a modification of an HPLC method reported by Foster et al. using a closely related structural analogue of zopiclone as internal standard. Zopiclone was detected at 306 nm and linear calibration curves were constructed in the range of 1.0-250 ng/mL for each enantiomer. The % CV at 2.5 ng/mL was 12.0% for (-)-zopiclone and 14.3% for (+)-zopiclone, and the limit of quantification of each enantiomer was 2.5 ng/mL. At higher concentrations, the coefficient of variation was less than 10%. The nominal concentration of quality control samples was predicted with an accuracy within a range of +/-11.6%. The method was used in the analysis of plasma obtained from psychiatric patients. One sample obtained following a non-fatal overdose with zopiclone contained the metabolites (-)-N-oxide zopiclone and both enantiomers of desmethyl zopiclone. The metabolite enantiomers were resolved on the column with retention times similar to zopiclone. The N-oxide metabolite co-eluted with internal standard. PMID- 11933025 TI - A rapid assay method for catechol-O-methyltransferase activity by flow injection analysis. AB - A rapid assay employing flow injection analysis (FIA) to determine the activity of purified catechol-O-methyltransferase (COMT) from porcine liver is described. The method was based on the determination of normetanephrine, the 3-O-methyl metabolite of the substrate norepinephrine. Excess norepinephrine was removed from the incubation mixture by alumina extraction twice to allow normetanephrine to be subjected to flow injection analysis, coulometrical oxidation, fluorogenic reaction with ethylenediamine and fluorescence detection. K(m) and V(max) values for COMT obtained with the system were 503 microM and 4.51 nmol/min/mg protein, respectively. The method is suitable for screening of COMT inhibitors or activators, as a large number of samples, up to 200, can be processed in one working day. PMID- 11933026 TI - The heterogeneity of the glycosylation of alpha-1-acid glycoprotein between the sera and synovial fluid in rheumatoid arthritis. AB - Alpha-1-acid glycoprotein (AGP) is a plasma glycoprotein produced by the liver that undergoes increased production and altered glycosylation in several physiological and pathological conditions including rheumatoid arthritis. To date, although present in the synovial fluid of rheumatoid arthritis patients, there has been no evidence for the separate extra-hepatic production of AGP. This study indicates that there could be a localized production of AGP in rheumatoid synovial fluid by demonstrating that the glycosylation patterns of AGP differed between the serum and synovial fluid in the same rheumatoid patient. Serum AGP was largely composed of fucosylated tri- and tetra-antennary oligosaccharide chains while the synovial fluid contained mainly bi-antennary chains that were fucosylated to a lesser extent. This structural heterogeneity of glycosylation resulted in functional diversity; serum but not synovial AGP is able to inhibit binding to the cell adhesion molecule E-selectin through expression of antigen sialyl Lewis X. PMID- 11933027 TI - Sensitive and simultaneous determination of HIV protease inhibitors in rat biological samples by liquid chromatography-mass spectrometry. AB - A sensitive and simultaneous liquid chromatographic-mass spectrometric (LC/MS) method for the determination of current four HIV protease inhibitors (PIs), indinavir (IDV), saquinavir (SQV), nelfinavir (NFV) and amprenavir (APV) in rat plasma and liver dialysate by a microdialysis method was described. An isocratic LC/MS method in combination with atmospheric pressure chemical ionization was developed for the determination of these four PIs in biological samples in the same run. The analytes including an internal standard were extracted from 100 microL of plasma or 150 microL of liver dialysate samples by salting-out with 100 microL of ice-cold 2 M K(3)PO(4) followed by ether extraction. The separation of analytes was carried out on a reversed-phase semi-micro column using 50% of acetonitrile containing 1% acetic acid as mobile phase at a flow rate of 0.2mL/min(-1). The separation was completed within 5 min. Precision, recovery and limits of detection indicated that the method was suitable for the quantitative determination of these PIs in rat plasma or liver dialysate. This simple, sensitive and highly specific LC/MS method is suitable for pharmacokinetic studies and therapeutic drug monitoring in AIDS patients who receive double protease therapy. PMID- 11933029 TI - Analysis of tofisopam in human serum by column-switching semi-micro high performance liquid chromatography and evaluation of tofisopam bioequivalency. AB - A rapid and sensitive column-switching semi-micro HPLC method is described for the direct analysis of tofisopam in human serum. The sample (100 microL) was directly injected onto the precolumn (Capcell Pak MF Ph-1), where unretained proteins were eluted to waste. Tofisopam was then eluted into an enrichment column using 13% acetonitrile in 50 mM phosphate buffer (pH 7.0) containing 5 mM sodium octanesulfonate and subsequently into the analytical column using 43% acetonitrile in 0.1% phosphoric acid containing 5 mM sodium octanesulfonate. The detection limit (2 ng/mL), good precision (CV < or = 4.2%) and speed (total analysis time 24 min) of the present method were sufficient for drug monitoring. This method was successfully applied to a bioequivalence test of two commercial tofisopam tablets. PMID- 11933028 TI - Rapid high-performance liquid chromatographic assay of dorzolamide in rabbit aqueous humor. AB - A rapid high-performance liquid chromatographic method using a C(18) reversed phase column (Hypersil ODS) with UV detection at 254 nm and a simple pre treatment of samples is presented for the analysis of dorzolamide, a carbonic anhydrase inhibitor, in rabbit aqueous humor. A water solution containing 2% ZnSO(4) small middle dot7H(2)O was used to deproteinize aqueous humor samples. The mobile phase consisted of 7% CH(3)CN and 93% of a solution containing 1% TEA adjusted to pH = 3.5 with H(3)PO(4). Paracetamol was found to be a suitable internal standard. The standard curves were linear in the detection range. The precision and the accuracy were <5% for both intra- and inter-day assays. PMID- 11933030 TI - High-performance liquid chromatographic method for determination of leucovorin in plasma: validation and application to a pharmacokinetic study in healthy volunteers. AB - A sensitive and reliable high-performance liquid chromatographic (HPLC) method, using a solid-phase extraction (SPE), was developed and validated for determination of leucovorin (LV) in human plasma. Plasma sample was extracted by using a Sep-Pak cartridge which could be renewable. The sample was analyzed by HPLC with UV detection at 286 nm. The method was shown to perform selectively and sensitively for LV. The main metabolite of LV, 5-methyltetrahydrofolic acid, and endogenous substances in plasma did not show any interference in the analysis. The limit of detection was 10 ng/mL for LV in plasma and the linear range was 50 1500 ng/mL in plasma. The relative standard deviation (RSD) of intra-day and inter-day assays was 2.8-6.1% and 2.4-5.3%, respectively. The extraction recoveries of LV in plasma were over 90%. The method was proved to be applicable to the pharmacokinetic study of LV in healthy volunteers after a single oral administration (75 mg). The pharmacokinetic parameters and relative bioavailability were investigated for domestic LV tablet and capsule vs an imported tablet. PMID- 11933031 TI - Capillary isoelectric focusing-mass spectrometry: analysis of protein mixtures from human body fluids. AB - Isoelectric focusing within a fused silica capillary (cIEF) has proved to be a powerful and practical method for high-resolution separation of analytes from complex biological mixtures. This technique overcomes many of the problems of isoelectric focusing within slab gel media. However current cIEF systems commonly utilize UV detection which limits the detail of analyte structural information that is obtained during analysis. The use of mass spectrometry (MS) as the detection system provides much greater structural information about the detected analytes allowing accurate relative molecular mass (M(r)) determination for proteins and polypeptides. We have constructed a cIEF-MS interface and compared the separation of standard proteins analyzed by cIEF-UV with cIEF-MS. This allowed rapid optimization of the cIEF-MS system performance. Further we have demonstrated the use of MS as a detection system provides accurate M(r) information and can provide analyte modification details. These factors increase the likelihood of absolute identification for physiological proteins within complex in vivo-derived mixtures. To demonstrate the value of cIEF-MS in such analyses we have undertaken an examination of cerebrospinal fluid (CSF), and tentatively identified a number of constituent proteins. We have also analyzed whole blood from control and diabetic patients. We show that glycated alpha- and beta- chains of hemoglobin are found in almost equal abundance in diabetic patient blood. From these results we suggest cIEF-MS is an efficient and useful tool for the separation and examination of in vivo-derived analytes within physiological fluids. PMID- 11933032 TI - Selection models and pattern-mixture models to analyse longitudinal quality of life data subject to drop-out. AB - Longitudinally observed quality of life data with large amounts of drop-out are analysed. First we used the selection modelling framework, frequently used with incomplete studies. An alternative method consists of using pattern-mixture models. These are also straightforward to implement, but result in a different set of parameters for the measurement and drop-out mechanisms. Since selection models and pattern-mixture models are based upon different factorizations of the joint distribution of measurement and drop-out mechanisms, comparing both models concerning, for example, treatment effect, is a useful form of a sensitivity analysis. PMID- 11933033 TI - Coping with missing data in clinical trials: a model-based approach applied to asthma trials. AB - In most clinical trials, some patients do not complete their intended follow-up according to protocol, for a variety of reasons, and are often described as having 'dropped out' before the conclusion of the trial. Their subsequent measurements are missing, and this makes the analysis of the trial's repeated measures data more difficult. In this paper we briefly review the reasons for patient drop-out, and their implications for some commonly used methods of analysis. We then propose a class of models for modelling both the response to treatment and the drop-out process. Such models are readily fitted in a Bayesian framework using non-informative priors with the software BUGS. The results from such models are then compared with the results of standard methods for dealing with missing data in clinical trials, such as last observation carried forward. We further propose the use of a time transformation to linearize an asymptotic pattern of repeated measures over time and therefore simplify the modelling. All these ideas are illustrated using data from a five-arm asthma clinical trial. PMID- 11933035 TI - Estimation of an errors-in-variables regression model when the variances of the measurement errors vary between the observations. AB - It is common in the analysis of aggregate data in epidemiology that the variances of the aggregate observations are available. The analysis of such data leads to a measurement error situation, where the known variances of the measurement errors vary between the observations. Assuming multivariate normal distribution for the 'true' observations and normal distributions for the measurement errors, we derive a simple EM algorithm for obtaining maximum likelihood estimates of the parameters of the multivariate normal distributions. The results also facilitate the estimation of regression parameters between the variables as well as the 'true' values of the observations. The approach is applied to re-estimate recent results of the WHO MONICA Project on cardiovascular disease and its risk factors, where the original estimation of the regression coefficients did not adjust for the regression attenuation caused by the measurement errors. PMID- 11933034 TI - Longitudinal profiles of health care providers. AB - Provider profiling is the activity of collecting, comparing and reporting quality of care measures for individuals, groups, agencies and institutions that provide health care services. Univariate provider profiles, such as hospital-specific mortality rates, have been constructed using cross-sectional data based on posterior summaries or maximum likelihood estimates. As data continue to be collected over time, the construction and interpretation of longitudinal profiles of health care providers will become increasingly important. Longitudinal series can be used to improve the precision of estimates - a feature that is particularly important for providers who treat a small number of patients per year. We extend and apply hierarchical models to examine and classify provider performance over time using two examples, one in the area of cardiology and the other in mental health. Performance is evaluated using the squared Mahalanobis distance and posterior probabilities based on this distance. By comparing providers based on level and temporal trend simultaneously, conservative but comprehensive assessments of performance are possible. Furthermore, the longitudinal profiles developed are easily interpreted and flexible, making them of practical use to policy-makers. PMID- 11933036 TI - Functional data analysis with application to periodically stimulated foetal heart rate data. I: functional regression. AB - Functional regression is used to model longitudinal data where the number of measurements on the functional covariate is much greater than the number of subjects in the study. Thus, functional regression can be thought of as singular longitudinal analysis. We have modified existing functional regression techniques to the case of a functional covariate with a repeated stimulus. We applied this modified functional regression to periodically stimulated foetal heart rates. The heart rate tracings were used as a predictor of the child's psychomotor development at approximately 18 months of age. In the past, this type of data has been analysed using the partially subjective concept of habituation. By using the entire heart rate tracings through functional regression, we have the advantage that habituation does not need to be defined and all available information is used to predict later child development. PMID- 11933037 TI - Functional data analysis with application to periodically stimulated foetal heart rate data. II: functional logistic regression. AB - We present a basis solution for the modelling of a binary response with a functional covariate plus any number of scalar covariates. This can be thought of as singular longitudinal data analysis as there are more measurements on the functional covariate than subjects in the study. The maximum likelihood parameter estimates are found using a basis expansion and a modified Fisher scoring algorithm. This technique has been extended to model a functional covariate with a repeated stimulus. We used periodically stimulated foetal heart rate tracings to predict the probability of a high risk birth outcome. It was found that these tracings could predict 94.1 per cent of the high risk pregnancies and without the stimulus, the heart rates were no more predictive than chance. PMID- 11933038 TI - Non-parametric confidence interval estimation for competing risks analysis: application to contraceptive data. AB - Non-parametric maximum likelihood estimation of the cause specific failure probability, and of its standard error, in the presence of competing risks is discussed with reference to some contraceptive use dynamics data from Bangladesh. The cause specific incidence function provides an intuitively appealing summary curve for failure rates and probabilities, such as probabilities of discontinuation of different kinds of contraception, based on right-censored data of the particular event. Dinse and Larson's formula can be utilized to calculate the standard error of the cause specific probability for the failure type of interest, and to construct pointwise confidence intervals. The accuracy of these intervals, as well as those based on the log(-log) transformation and the arcsine transformation, are compared by simulations. We find that Dinse and Larson's formula, used in conjuction with a log(-log) transform, yields reliable standard error estimates and accurate coverage in samples of small and large size, and can be recommended for use in this situation. PMID- 11933039 TI - Prediction trees with soft nodes for binary outcomes. AB - Consider the problem of predicting the occurrence of an event, the onset of diabetes mellitus, say, from a vector of continuous and discrete predictors. We propose a new algorithm for the construction of a tree-structured predictor for the event of interest, which uses a new approach for dealing with continuous predictors. The novelty is that the tree uses splits for continuous variables. This means that at each node an individual goes to the right branch with a certain probability, function of a predictor. The predictor as well as the particular shape of the function is chosen from the data by the proposed algorithm. We evaluate its performance on several real data sets, in particular comparing it with a standard tree-growing algorithm. We also present an analysis of a well-known data set, the Pima Indian diabetes data set, to illustrate the application of the method in biostatistics. PMID- 11933040 TI - The impact of covariate imbalance on the size of the logrank test in randomized clinical trials by N. Kinukawa, T. Nakamura, K. Akazawa and Y. Nose, Statistics in Medicine 2000; 19: 1955-1967. PMID- 11933042 TI - Heterogeneity in phase I clinical trials: prior elicitation and computation using the continual reassessment method by A. Legedza and J. G. Ibrahim, Statistics in Medicine 2001; 20: 867-882. PMID- 11933043 TI - Promoting axonal regeneration in the central nervous system by enhancing the cell body response to axotomy. AB - Neurons projecting into the peripheral nervous system (PNS) regenerate their axons after injury, in contrast to those confined to the central nervous system (CNS). Both neuronal and nonneuronal factors contribute to the lack of CNS regeneration. In this review we concentrate on the differential gene expression response to axotomy in PNS vs. CNS neurons. In general CNS neurons fail to up regulate or sustain the expression of regeneration-associated proteins (RAGs), including trophic factors and their receptors. The presumed lack of trophic support of axotomized CNS neurons provided the rationale for the exogenous application of trophic factors, either to the lesion site or to the cell bodies. Here, we review our data on the application of trophic factors to rubrospinal and corticospinal neurons. Cell body treatment of axotomized rubrospinal neurons with brain-derived neurotrophic factor (BDNF) reversed atrophy, increased GAP-43 and Talpha-1 tubulin mRNA expression, and promoted axonal regeneration into peripheral nerve grafts. Importantly, BDNF cell body treatment was still effective in the chronic setting, i.e., when initiated 1 year after injury, but BDNF had no effect when applied to the chronic spinal cord injury site. The ability to promote regeneration in chronically injured neurons will hopefully contribute to the development of treatment strategies for chronic spinal injuries. PMID- 11933044 TI - Efficacy of methylprednisolone therapy for the injured rat spinal cord. AB - Currently the synthetic glucocorticosteroid methylprednisolone sodium succinate (MPSS) is the standard therapy after acute spinal cord injury (SCI) in humans based on reported neurological improvements. The mechanisms for its beneficial actions are not entirely clear, but experimental evidence suggests MPSS affords some degree of neuroprotection. As many studies with rat models of SCI have been unable to demonstrate improved behavioral outcome or tissue sparing after MPSS treatment, we chose to stereologically assess whether it alters lesion volume and tissue sparing over time, as well as long-term behavioral recovery. Adult rats subjected to contusion SCI with the NYU impactor were administered either MPSS or saline for 24 hr beginning 5 min post injury. Over time the lesion dimensions were extremely dynamic, such that by 6 weeks post injury the volumes were reduced to a third of those seen after the first week. MPSS marginally reduced lesion volumes across time vs. controls, but the amount of spared gray and white matter remained unaltered between the two groups. Behavioral results further showed that MPSS failed to improve recovery of hind-limb function. These findings add to the emerging scrutiny of MPSS as the standard therapy for acute SCI, as well as indicate the existence of a therapeutic window for tissue sparing restricted to the first several days after this type of SCI in rats. Equally important, our results caution the use of lesion volume dimensions or percent tissue sparing at the epicenter as indicators of therapeutic efficacy because neither reflects the actual amount of tissue sparing. PMID- 11933045 TI - Study of expression of myelin basic proteins (MBPs) in developing rat brain using a novel antibody reacting with four major isoforms of MBP. AB - Myelin basic proteins (MBPs) are the major protein components of myelin. MBP isoforms are known to have different expression patterns. In order to distinguish the different expression patterns on myelination, we have developed a novel antibody reacting with the four major isoforms of MBPs with molecular masses of 21.5 kDa, 18.5 kDa, 17.0 kDa, and 14.0 kDa. These MBPs were initially separated by acid urea gel and sodium dodecyl sulfate polyacrylamide gel electrophoreses and detected with the luminol reaction. Then the antibody developed was used to determine the relative amounts of MBP isoforms. The MBPs of oligodendrocytes were detected by the enhanced luminol reaction using Renaissance (Dupont NEN, Boston, MA). From the immunological aspect, the MBP monoclonal antibody (Sires et al. [1981] Science 214:87-89) was revealed to recognize MBPs with molecular masses of 21.5 kDa and 18.5 kDa. Furthermore, we found that Ile-166 in the rat 18.5-kDa MBP isomers was replaced by methionine. The 14.0-kDa and 18.5-kDa isoforms of MBP are the most abundant MBP species and comprise more than 70% of the total MBPs in 3.5 and 24-month-old rats. MBPs are expressed during development and the compositions of MBPs in mature (3.5 months old) and aged (24 months old) rats were almost the same. The expression of the 14.0-kDa and 18.5-kDa MBPs occurred earlier in the cerebellum and the spinal cord than in the cerebrum by approximately 1 week. MBPs are also expressed upon oligodendrocyte maturation by interacting with astrocytes. The above results suggest that the regulation of MBP isoforms during development and oligodendrocyte differentiation may indicate the point of occurrence of both the unique patterns of isoform expression and the shift in intracellular localization of MBPs with the maturation of oligodendrocytes. PMID- 11933046 TI - Slow axonal transport of the cytosolic chaperonin CCT with Hsc73 and actin in motor neurons. AB - Molecular chaperones are well known for their role in facilitating the folding of nascent and newly synthesized proteins, but have other roles, including the assembly, translocation and renaturation of intracellular proteins. Axons are convenient tissues for the study of some of these other roles because they lack the capacity for significant protein synthesis. We examine the axonal transport of the cytosolic chaperonin containing T- complex polypeptide 1 (CCT) by labeling lumbar motor neurons with [35S]methionine and examining sciatic nerve proteins by 2-D gel electrophoresis and immunoblotting. All CCT subunits identifiable with specific antibodies, namely CCTalpha, CCTbeta, CCTgamma and CCTepsilon/CCTtheta; (the latter two subunits colocalized in analyses of rat nerve samples), appeared to be labeled in "slow component b" of axonal transport along with the molecular chaperone Hsc73 and actin, a major folding substrate for CCT. Our results are consistent with molecular chaperones having a post-translational role in maintaining the native form of actin during its slow transport to the axon terminal and ensuring its correct assembly into microfilaments. PMID- 11933047 TI - Constitutive expression of heterologous N-methyl-D-aspartate receptor subunits in rat adrenal medulla. AB - An RT-PCR analysis revealed constitutive expression of mRNA for N-methyl-D aspartate (NMDA) receptor (NR)-1, NR-2C, and NR-2D subunits in rat adrenal and pituitary glands, in addition to brain and retina. Constitutive expression of mRNA was detected for the NR-2A subunit in pituitary but not adrenal gland. Although on Western blotting assays adrenal medulla exhibited expression of NR-1 subunit protein without expression of NR-2C and NR-2D subunit proteins, an immunohistochemical investigation clearly showed selective localization of proteins for NR-1, NR-2C, and NR-2D subunits in adrenal medulla but not in adrenal cortex. Prior treatment with different glycosidases invariably resulted in a marked increase in immunoreactivity to the anti-NR-1 antibody in both hippocampus and adrenal medulla. An intraperitoneal injection of a blocker of NMDA receptor channel, but not NMDA itself, led to marked potentiation of DNA binding activity of the transcription factor activator protein-1 in adrenal nuclear extracts 2 hr after administration. These results suggest that heteromeric NMDA receptor channels may be constitutively and functionally expressed with glycosylation of NR-1 subunit under the influence of tonic stimulation by circulating agonists such as L-glutamate in rat adrenal medulla. PMID- 11933049 TI - Modulation of 5-HT system in mice with a targeted disruption of neuromedin B receptor. AB - To assess the role of neuromedin B receptor (NMB-R) on the modulation of serotonergic (5-HT) system, the function of the 5-HT system was examined in mice lacking the NMB-R gene. Immunohistochemical analysis of brain sections revealed that 5-HT expression level in the dorsal raphe neurons was elevated in NMB-R deficient mice compared with wild-type mice. Although restraint stress enhanced 5 HT expression in these neurons in wild-type mice, this treatment did not affect 5 HT expression level in NMB-R-deficient mice, indicating the modulation of 5-HT system in the mutant mice. Since 5-HT system is involved in responses to stress and anxiety, we characterized stress response in these mice. The number of c-Fos expressing cells in the paraventricular nucleus of the hypothalamus was higher in NMB-R-deficient mice than in wild-type mice in both basal and stressed conditions. Moreover, the plasma corticosterone level under restraint stress was elevated in NMB-R-deficient mice compared to wild-type mice. In the forced swimming tests, the duration of immobility was longer in mutant mice than in wild type mice. These data show dysregulated response to stress in NMB-R-deficient mice. However, behavior related to anxiety, assessed by elevated plus-maze and light-dark box, was not affected in NMB-R-deficient mice. NMB-R is known to be expressed in dorsal raphe neurons, and our data suggest that NMB-R has an important role in fine tuning of subsets of 5-HT neurons in this nucleus, and impairment of this system leads to the dysregulated response to stress. PMID- 11933048 TI - Establishment and characterization of immortalized neuronal cell lines derived from the spinal cord of normal and trisomy 16 fetal mice, an animal model of Down syndrome. AB - We report the establishment of continuously growing cell lines from spinal cords of normal and trisomy 16 fetal mice. We show that both cell lines, named M4b (derived from a normal animal) and MTh (trisomic) possess neurological markers by immunohistochemistry (neuron specific enolase, synaptophysin, microtubule associated protein-2 [MAP-2], and choline acetyltransferase) and lack glial traits (glial fibrillary acidic protein and S100). MTh cells were shown to overexpress mRNA of Cu/Zn superoxide dismutase, whose gene is present in autosome 16. We also studied intracellular Ca2+ signals ([Ca2+]i) induced by different agonists in Indo-1 loaded cells. Basal [Ca2+]i was significantly higher in MTh cells compared to M4b cells. Glutamate (200 microM) and (1S,3R)-1 aminocyclopentane-1,3-dicarboxylic acid (ACDP) (100 microM) induced rapid, transient increases in [Ca2+]i in M4b and MTh cells, indicating the presence of glutamatergic metabotropic receptors. N-methyl-D-aspartate (NMDA) and kainate, but not alpha-amino-hydroxy-5-methylisoxazole-4-propionic acid (AMPA), produced [Ca2+)]i rises in both cell types. MTh cells exhibited faster time-dependent decay phase kinetics in glutamate-induced responses compared to M4b cells. Nicotine induced a transient increase in [Ca2+]i in M4b and MTh cells, with significantly greater amplitudes in the latter compared to the former. Further, both cell types responded to noradrenaline. Finally, we examined cholinergic function in both cell lines and found no significant differences in the [3H] choline uptake, but fractional acetylcholine release induced by either K+, glutamate or nicotine was significantly higher in MTh cells. These results show that M4b and MTh cells have neuronal characteristics and the MTh line shows differences which could be related to neuronal pathophysiology in Down's syndrome. PMID- 11933050 TI - Regional distribution of heme oxygenase, HSP70, and glutathione in brain: relevance for endogenous oxidant/antioxidant balance and stress tolerance. AB - It is generally recognized that lipid peroxides play an important role in the pathogenesis of several diseases and that sulfhydryl groups are critically involved in cellular defense against endogenous or exogenous oxidants. Recent evidence indicates that lipid peroxides directly participate in induction of cytoprotective proteins, such as heat shock proteins (Hsps), which play a central role in the cellular mechanisms of stress tolerance. Heme oxygenase (HO) is a stress protein that has been implicated in defense mechanisms against agents that may induce oxidative injury, such as endotoxins, cytokines and heme and its induction represents a common feature in a number of neurodegenerative diseases. In the present report we studied regional distribution of heme oxygenase (HO) activity and protein expression, together with that of Hps70, in brain of C57BL6 mice. Endogenous lipid peroxidation was investigated on the basis of the analysis of ultra weak chemiluminescence, hydro peroxides and lipid soluble fluorescent products, and compared to the regional distribution of thiols, antioxidant enzymes and trace metals. Our results show that levels of HO activity and expression of inducible Hsp70 and the ratio of GSH/GSSG in the different brain regions examined were positively correlated with the content of peroxides. Substantia Nigra was the brain area exhibiting the highest levels of HO-2, constitutive and inducible Hsp70, GSSG, peroxides, iron, and calcium, in contrast with the lowest content in GSH, GSH/GSSG ratio and glutathione reductase activity, compared to the other cerebral regions examined. Among these, cortex showed the lowest levels of HO-2, Hsp70, GSSG and peroxides that were associated with the highest levels of GSH and GSH/GSSG ratio. These data support the hypothesis that the glutathione redox state and basal peroxides can directly participate in the signaling pathways of heat shock protein expression and hence of stress tolerance. PMID- 11933051 TI - Effect of the iron chelator desferrioxamine on manganese-induced toxicity of rat pheochromocytoma (PC12) cells. AB - Alterations in iron levels are likely to influence the biological actions of Mn in PC12 cells, because both metals are transported via the divalent metal transporter 1 (DMT1; also Nramp2 or DCT1). Studies were performed to determine the effect of the iron chelator desferrioxamine (DfO) on Mn-induced PC12 cell death and neuronal differentiation. Cell death almost doubled when PC12 cells were exposed for 24 hr to both DfO (10 microM) and Mn (0.3 mM) as opposed to Mn alone. DfO also stimulated Mn-induced neuronal differentiation by enhancing the phosphorylation of both ERK1 and 2 and also attenuated the increase in caspase 3 like activity induced by 0.3 mM Mn by approximately 50%, indicating that caspase activation, as reported previously, does not contribute to Mn-induced PC12 cell death. DfO also affected Mn-induced suppression of mitochondrial function as indicated by an additional 16% loss of ATP formation in PC12 cells cotreated with 0.3 mM Mn. Because sequestration of iron by DfO would be expected to lead to increased transport of Mn, studies were performed to determine whether iron inhibited Mn transport in PC12 cells. Iron inhibited 54Mn transport with an IC50 of approximately 20 microM. In addition, coincubation of DfO with Mn in PC12 cells resulted in increased expression of both the iron response element-positive and the iron response element-negative forms of DMT1. Taken together, these results demonstrate that iron status is likely to have a direct effect on the uptake and biological actions of Mn and probably other divalent metals that are transported by DMT1. PMID- 11933052 TI - Gender-dependent hypoxic tolerance mediated via gender-specific mechanisms. AB - Primary hypoxic tolerance and preconditioning are gender dependent and modulated in females during the estrus cycle. The underlying mechanisms, however, remain to be determined. mRNA of estrogen receptor-alpha (EAR), progesterone receptor (PR), and adenosine receptor subtypes A1 and A3 (A1R and A3R) were investigated with reverse transcriptase-PCR in hippocampi from control male and female mice and animals treated in vivo with a single i.p. injection of 20 mg/kg body weight 3 nitropropionate (3NP) 1 or 24 hr prior to preparation. Results were analyzed relative to expression in hippocampi from untreated males. mRNA levels of EAR and A1R were alike in males and females and unaltered by preconditioning with 3NP. In contrast, PR mRNA levels were alike in males and females during proestrus but lower during estrus and diestrus (85% +/- 15%, P < 0.05; and 80% +/- 10%, P < 0.05, respectively). Upon preconditioning, PR mRNA decreased to 67% +/- 19% (P < 0.05) and 56% +/- 13% (P < 0.05) during proestrus and diestrus, respectively, but was unaltered during estrus and in males. On preconditioning, A3R mRNA decreased from 115% +/- 16% to 86% +/- 29% (P < 0.05) during diestrus but remained at the control level during proestrus and estrus. With low-level expression of PRs, as achieved upon preconditioning, hypoxic tolerance is increased. Other than in males, adenosine A3 receptors are not up-regulated upon preconditioning in females. Thus, not only is net hypoxic tolerance gender dependent but mechanisms conferring hypoxic tolerance are gender specific. PMID- 11933053 TI - beta-Amyloid peptide induces ultrastructural changes in synaptosomes and potentiates mitochondrial dysfunction in the presence of ryanodine. AB - In Alzheimer's disease (AD), loss of synapses exceeds neuronal loss and some evidence suggests a role of beta-amyloid protein (Abeta) in synaptic degeneration through a mechanism which may involve intraneuronal Ca2+ dyshomeostasis. Emerging evidence points to the participation of the internal Ca2+ stores in the pathophysiology of neurodegeneration in AD. To test the involvement of intrasynaptic Ca2+ mobilization in A toxicity, we explored the role of ryanodine receptor activation in rat cortical synaptosomes taken as a model system for the central presynapses. Evaluation of synaptosomal mitochondrial redox capacity was assessed by the MTT reduction technique, and ultrastructural changes of synaptosomes after exposure to Abeta and ryanodine were evaluated by electron microscopy. Our results show that Abeta potentiates mitochondrial dysfunction in the presence of ryanodine and induces morphological changes consisting of mitochondrial swelling and intense small synaptic vesicles depletion. These changes were accompanied by a reduction in the content of synaptophysin and actin proteins. The reduction of actin immunoreactivity was reversed in the presence of a wide range caspase inhibitors, suggesting the activation of synaptic apoptotic mechanisms. PMID- 11933054 TI - Synucleins in glaucoma: implication of gamma-synuclein in glaucomatous alterations in the optic nerve. AB - Synucleins are small proteins associated with neurodegenerative diseases and some forms of cancer. They are studied predominantly in the brain; information about their presence and functions in ocular tissues is scarce. Here we describe the localization of three members of the synuclein family in the optic nerve of donors with different types of glaucoma compared with control samples from donors without ocular diseases. We did not find significant differences in the localization of alpha- and beta-synucleins in the optic nerve or retina of glaucoma patients compared with controls, whereas considerable redistribution of gamma-synuclein occurred in the glaucomatous optic nerve compared with control eye without glaucoma. In the optic nerve from control and glaucomatous individuals, nerve bundles are immunopositive for gamma-synuclein; however, a strong gamma-synuclein-immunopositive staining in a subset of glial cells was observed in the lamina and postlamina cribrosa regions of the optic nerve only in glaucoma patients. In the optic nerve of rats with episcleral vein cauterization used as an animal model of glaucoma, the quantity of both gamma-synuclein mRNA and protein was decreased compared with the optic nerves of control animals. Incubation of rat astrocyte culture at elevated hydrostatic pressure reduced the amount of gamma-synuclein but did not affect the quantities of actin and glial fibrillary acidic protein. These data suggest that significant changes in the pattern of expression and/or localization occur in the glaucomatous optic nerve for gamma-synuclein but not for alpha- and beta-members of the synuclein family. PMID- 11933055 TI - Differential enhancement of dialysate serotonin levels in distinct brain regions of the awake rat by modafinil: possible relevance for wakefulness and depression. AB - The present in vivo microdialysis study evaluates the possible existence of a differential regulation of serotonergic transmission by the antinarcoleptic drug modafinil [(diphenyl-methyl)-sulfinyl-2-acetamide; Modiodal] among various brain regions of the awake rat. The results show that, in the cerebral cortex, the central amygdala, and the dorsal raphe nucleus, modafinil in the dose range of 10 100 mg/kg i.p. dose-dependently increases dialysate serotonin (5-HT) levels. In other brain areas, such as the medial preoptic area and the posterior hypothalamus, the modafinil-induced increase in dialysate 5-HT levels is observed only at tenfold higher doses (100 mg/kg), 10-30 mg/kg being ineffective. Together these data suggest that, in the frontal cortex, the amygdala, and the dorsal raphe, modafinil is more potent in enhancing extracellular 5-HT levels and presumably 5-HT transmission than in the medial preoptic area and the posterior hypothalamus. In view of the role of ascending 5-HT pathways in arousal and depression, it seems likely that the antinarcoleptic drug modafinil may also have an antidepressant potential in addition to its wakefulness-promoting action, both actions involving enhancement of 5-HT neurotransmission. PMID- 11933056 TI - Myelin: development, disorders and neural repair. A satellite meeting of the International and American Societies for Neurochemistry joint meeting in Buenos Aires. Bastion del Carmen, September 1-4, 2001, Colonia, Uruguay. PMID- 11933057 TI - Folding of a small helical protein using hydrogen bonds and hydrophobicity forces. AB - A reduced protein model with five to six atoms per amino acid and five amino acid types is developed and tested on a three-helix-bundle protein, a 46-amino acid fragment from staphylococcal protein A. The model does not rely on the widely used Go approximation, which ignores non-native interactions. We find that the collapse transition is considerably more abrupt for the protein A sequence than for random sequences with the same composition. The chain collapse is found to be at least as fast as helix formation. Energy minimization restricted to the thermodynamically favored topology gives a structure that has a root-mean-square deviation of 1.8 A from the native structure. The sequence-dependent part of our potential is pairwise additive. Our calculations suggest that fine-tuning this potential by parameter optimization is of limited use. PMID- 11933058 TI - Classification of proteins based on the properties of the ligand-binding site: the case of adenine-binding proteins. AB - Comparative analysis of protein binding sites for similar ligands yields information about conserved interactions, relevant for ligand affinity, and variable interactions, which are important for specificity. The pattern of variability can indicate new targets for a pharmacologically validated class of compounds binding to a similar site. A particularly vast group of therapeutically interesting proteins using the same or similar substrates are those that bind adenine-containing ligands. Drug development is focusing on compounds occupying the adenine-binding site and their specificity is an issue of paramount importance. We use a simple scheme to characterize and classify the adenine binding sites in terms of their intermolecular interactions, and show that this classification does not necessarily correspond to protein classifications based on either sequence or structural similarity. We find that only a limited number of the different hydrogen bond patterns possible for adenine-binding is used, which can be utilized as an effective classification scheme. Closely related protein families usually share similar hydrogen patterns, whereas non-polar interactions are less well conserved. Our classification scheme can be used to select groups of proteins with a similar ligand-binding site, thus facilitating the definition of the properties that can be exploited to design specific inhibitors. PMID- 11933059 TI - Probing the structure of the warfarin-binding site on human serum albumin using site-directed mutagenesis. AB - The binding of warfarin to the following human serum albumin (HSA) mutants was examined: K195M, K199M, F211V, W214L, R218M, R222M, H242V, and R257M. Warfarin bound to human serum albumin (HSA) exhibits an intrinsic fluorescence that is approximately 10-fold greater than the corresponding signal for warfarin in aqueous solution. This property of the warfarin/HSA complex has been widely used to determine the dissociation constant for the interaction. In the present study, such a technique was used to show that specific substitutions in subdomain 2A altered the affinity of HSA for warfarin. The fluorescence of warfarin/mutant HSA complexes varied widely from the fluorescence of the warfarin/wild-type HSA complex at pH = 7.4, suggesting changes in the structure of the complex resulting from specific substitutions. The fluorescence of the warfarin/wild-type HSA complex increases about twofold as the pH is increased from 6.0 to 9.0 due to the neutral-to-base (N-B) transition, a conformational change that occurs in HSA as a function of pH. Changes in the fluorescence of warfarin/mutant HSA complexes as a function of pH suggests novel behavior for most HSA species examined. For the HSA mutants F211V and H242V, the midpoint of the N-B transition shifts from a wild type pH of 7.8 to a pH value of 7.1-7.2. PMID- 11933060 TI - Structural genomics analysis: characteristics of atypical, common, and horizontally transferred folds. AB - We conducted a structural genomics analysis of the folds and structural superfamilies in the first 20 completely sequenced genomes by focusing on the patterns of fold usage and trying to identify structural characteristics of typical and atypical folds. We assigned folds to sequences using PSI-blast, run with a systematic protocol to reduce the amount of computational overhead. On average, folds could be assigned to about a fourth of the ORFs in the genomes and about a fifth of the amino acids in the proteomes. More than 80% of all the folds in the SCOP structural classification were identified in one of the 20 organisms, with worm and E. coli having the largest number of distinct folds. Folds are particularly effective at comprehensively measuring levels of gene duplication, because they group together even very remote homologues. Using folds, we find the average level of duplication varies depending on the complexity of the organism, ranging from 2.4 in M. genitalium to 32 for the worm, values significantly higher than those observed based purely on sequence similarity. We rank the common folds in the 20 organisms, finding that the top three are the P-loop NTP hydrolase, the ferrodoxin fold, and the TIM-barrel, and discuss in detail the many factors that affect and bias these rankings. We also identify atypical folds that are "unique" to one of the organisms in our study and compare the characteristics of these folds with the most common ones. We find that common folds tend be more multifunctional and associated with more regular, "symmetrical" structures than the unique ones. In addition, many of the unique folds are associated with proteins involved in cell defense (e.g., toxins). We analyze specific patterns of fold occurrence in the genomes by associating some of them with instances of horizontal transfer and others with gene loss. In particular, we find three possible examples of transfer between archaea and bacteria and six between eukarya and bacteria. We make available our detailed results at http://genecensus.org/20. PMID- 11933061 TI - Prediction of coordination number and relative solvent accessibility in proteins. AB - Knowing the coordination number and relative solvent accessibility of all the residues in a protein is crucial for deriving constraints useful in modeling protein folding and protein structure and in scoring remote homology searches. We develop ensembles of bidirectional recurrent neural network architectures to improve the state of the art in both contact and accessibility prediction, leveraging a large corpus of curated data together with evolutionary information. The ensembles are used to discriminate between two different states of residue contacts or relative solvent accessibility, higher or lower than a threshold determined by the average value of the residue distribution or the accessibility cutoff. For coordination numbers, the ensemble achieves performances ranging within 70.6-73.9% depending on the radius adopted to discriminate contacts (6A 12A). These performances represent gains of 16-20% over the baseline statistical predictor, always assigning an amino acid to the largest class, and are 4-7% better than any previous method. A combination of different radius predictors further improves performance. For accessibility thresholds in the relevant 15-30% range, the ensemble consistently achieves a performance above 77%, which is 10 16% above the baseline prediction and better than other existing predictors, by up to several percentage points. For both problems, we quantify the improvement due to evolutionary information in the form of PSI-BLAST-generated profiles over BLAST profiles. The prediction programs are implemented in the form of two web servers, CONpro and ACCpro, available at http://promoter.ics.uci.edu/BRNN-PRED/. PMID- 11933062 TI - Role of hydrophilic and hydrophobic contacts in folding of the second beta hairpin fragment of protein G: molecular dynamics simulation studies of an all atom model. AB - Predicting the folding mechanism of the second beta-hairpin fragment of the Ig binding domain B of streptococcal protein G is unexpectedly challenging for simplified reduced models because the models developed so far indicated a different folding mechanism from what was suggested from high-temperature unfolding and equilibrium free-energy surface analysis based on established all atom empirical force fields in explicit or implicit solvent. This happened despite the use of empirical residue-based interactions, multibody hydrophobic interactions, and inclusions of hydrogen bonding effects in the simplified models. This article employs a recently developed all-atom (except nonpolar hydrogens) model interacting with simple square-well potentials to fold the peptide fragment by molecular dynamics simulation methods. In this study, 193 out of 200 trajectories are folded at two reduced temperatures (3.5 and 3.7) close to the transition temperature T* approximately 4.0. Each simulation takes <7 h of CPU time on a Pentium 800-MHz PC. Folding of the new all-atom model is found to be initiated by collapse before the formation of main-chain hydrogen bonds. This verifies the mechanism proposed from previous all-atom unfolding and equilibrium simulations. The new model further predicts that the collapse is initiated by two nucleation contacts (a hydrophilic contact between D46 and T49 and a hydrophobic contact between Y45 and F52), in agreement with recent NMR measurements. The results suggest that atomic packing and native contact interactions play a dominant role in folding mechanism. PMID- 11933063 TI - Identification of a novel class in the alpha/beta hydrolase fold superfamily: the N-myc differentiation-related proteins. AB - The alpha/beta hydrolases constitute a large protein superfamily that mainly consists of enzymes that catalyze a diverse range of reactions. These proteins exhibit the alpha/beta hydrolase fold, the essential features of which have recently been delineated: the presence of at least five parallel beta-strands, a catalytic triad in a specific order (nucleophile-acid-histidine), and a nucleophilic elbow. Because of the difficulties experimentally in identifying protein structures, we have used a Bayesian computational algorithm (PROBE) to identify the members of this superfamily based on distant sequence relationships. We found that the presence of five sequence motifs, which contain residues important for substrate binding and stabilization of the fold, are required for membership in this superfamily. The superfamily consists of at least 909 members, including the N-myc downstream regulated proteins, which are believed to be involved in cell differentiation. Unlike most of the other superfamily members, the N-myc downstream regulated proteins have never been proposed to possess the alpha/beta hydrolase fold and do not appear to be hydrolases. PMID- 11933064 TI - Hamming distance geometry of a protein conformational space: application to the clustering of a 4-ns molecular dynamics trajectory of the HIV-1 integrase catalytic core. AB - Protein structures can be encoded into binary sequences (Gabarro-Arpa et al., Comput Chem 2000;24:693-698) these are used to define a Hamming distance in conformational space: the distance between two different molecular conformations is the number of different bits in their sequences. Each bit in the sequence arises from a partition of conformational space in two halves. Thus, the information encoded in the binary sequences is also used to characterize the regions of conformational space visited by the system. We apply this distance and their associated geometric structures to the clustering and analysis of conformations sampled during a 4-ns molecular dynamics simulation of the HIV-1 integrase catalytic core. The cluster analysis of the simulation shows a division of the trajectory into two segments of 2.6 and 1.4 ns length, which are qualitatively different: the data points to the fact that equilibration is only reached at the end of the first segment. The Hamming distance is compared also to the r.m.s. deviation measure. The analysis of the cases studied so far shows that under the same conditions the two measures behave quite differently, and that the Hamming distance appears to be more robust than the r.m.s. deviation. PMID- 11933065 TI - Threading with chemostructural restrictions method for predicting fold and functionally significant residues: application to dipeptidylpeptidase IV (DPP IV). AB - We present a new method for more accurate modeling of protein structure, called threading with chemostructural restrictions. This method addresses those cases in which a target sequence has only remote homologues of known structure for which sequence comparison methods cannot provide accurate alignments. Although remote homologues cannot provide an accurate model for the whole chain, they can be used in constructing practically useful models for the most conserved-and often the most interesting-part of the structure. For many proteins of interest, one can suggest certain chemostructural patterns for the native structure based on the available information on the structural superfamily of the protein, the type of activity, the sequence location of the functionally significant residues, and other factors. We use such patterns to restrict (1) a number of possible templates, and (2) a number of allowed chain conformations on a template. The latter restrictions are imposed in the form of additional template potentials (including terms acting as sequence anchors) that act on certain residues. This approach is tested on remote homologues of alpha/beta-hydrolases that have significant structural similarity in the positions of their catalytic triads. The study shows that, in spite of significant deviations between the model and the native structures, the surroundings of the catalytic triad (positions of C(alpha) atoms of 20-30 nearby residues) can be reproduced with accuracy of 2-3 A. We then apply the approach to predict the structure of dipeptidylpeptidase IV (DPP-IV). Using experimentally available data identifying the catalytic triad residues of DPP-IV (David et al., J Biol Chem 1993;268:17247-17252); we predict a model structure of the catalytic domain of DPP-IV based on the 3D fold of prolyl oligopeptidase (Fulop et al., Cell 1998;94:161-170) and use this structure for modeling the interaction of DPP-IV with inhibitor. PMID- 11933066 TI - Contributions to the binding free energy of ligands to avidin and streptavidin. AB - The free energy of binding of a ligand to a macromolecule is here formally decomposed into the (effective) energy of interaction, reorganization energy of the ligand and the macromolecule, conformational entropy change of the ligand and the macromolecule, and translational and rotational entropy loss of the ligand. Molecular dynamics simulations with implicit solvation are used to evaluate these contributions in the binding of biotin, biotin analogs, and two peptides to avidin and streptavidin. We find that the largest contribution opposing binding is the protein reorganization energy, which is calculated to be from 10 to 30 kcal/mol for the ligands considered here. The ligand reorganization energy is also significant for flexible ligands. The translational/rotational entropy is 4.5-6 kcal/mol at 1 M standard state and room temperature. The calculated binding free energies are in the correct range, but the large statistical uncertainty in the protein reorganization energy precludes precise predictions. For some complexes, the simulations show multiple binding modes, different from the one observed in the crystal structure. This finding is probably due to deficiencies in the force field but may also reflect considerable ligand flexibility. PMID- 11933067 TI - Interhelical hydrogen bonds and spatial motifs in membrane proteins: polar clamps and serine zippers. AB - Polar and ionizable amino acid residues are frequently found in the transmembrane (TM) regions of membrane proteins. In this study, we show that they help to form extensive hydrogen bond connections between TM helices. We find that almost all TM helices have interhelical hydrogen bonding. In addition, we find that a pair of contacting TM helices is packed tighter when there are interhelical hydrogen bonds between them. We further describe several spatial motifs in the TM regions, including "Polar Clamp" and "Serine Zipper," where conserved Ser residues coincide with tightly packed locations in the TM region. With the examples of halorhodopsin, calcium-transporting ATPase, and bovine cytochrome c oxidase, we discuss the roles of hydrogen bonds in stabilizing helical bundles in polytopic membrane proteins and in protein functions. PMID- 11933068 TI - In silico two-hybrid system for the selection of physically interacting protein pairs. AB - Deciphering the interaction links between proteins has become one of the main tasks of experimental and bioinformatic methodologies. Reconstruction of complex networks of interactions in simple cellular systems by integrating predicted interaction networks with available experimental data is becoming one of the most demanding needs in the postgenomic era. On the basis of the study of correlated mutations in multiple sequence alignments, we propose a new method (in silico two hybrid, i2h) that directly addresses the detection of physically interacting protein pairs and identifies the most likely sequence regions involved in the interactions. We have applied the system to several test sets, showing that it can discriminate between true and false interactions in a significant number of cases. We have also analyzed a large collection of E. coli protein pairs as a first step toward the virtual reconstruction of its complete interaction network. PMID- 11933070 TI - Comparative structural analysis of psychrophilic and meso- and thermophilic enzymes. AB - Enzymes adapted to cold display structures comparable with those of their meso- and thermophilic homologs but are characterized by a higher catalytic efficiency at low temperatures and by thermolability at moderate temperatures. To identify the structural factors responsible of such features, we undertook a systematic comparative analysis of several structural properties in a data set consisting of 7 cold active enzymes belonging to different structural families and 28 related structures from meso/thermophiles representing most of the structural information now available. Only high-resolution and high-quality structures were considered. Properties were calculated and then compared for each pair of 3D structures displaying different temperatures of adaptation using a temperature-weighting scheme. The significance of the resulting differences was evaluated with a statistical method. Results reveal that each protein family adopts different structural strategies to adapt to low temperatures. However, some common trends are observed: the number of ion pairs, the side-chain contribution to the exposed surface, and the apolar fraction of the buried surface show a consistent decrease with decreasing optimal temperatures. PMID- 11933069 TI - Improving the prediction of protein secondary structure in three and eight classes using recurrent neural networks and profiles. AB - Secondary structure predictions are increasingly becoming the workhorse for several methods aiming at predicting protein structure and function. Here we use ensembles of bidirectional recurrent neural network architectures, PSI-BLAST derived profiles, and a large nonredundant training set to derive two new predictors: (a) the second version of the SSpro program for secondary structure classification into three categories and (b) the first version of the SSpro8 program for secondary structure classification into the eight classes produced by the DSSP program. We describe the results of three different test sets on which SSpro achieved a sustained performance of about 78% correct prediction. We report confusion matrices, compare PSI-BLAST to BLAST-derived profiles, and assess the corresponding performance improvements. SSpro and SSpro8 are implemented as web servers, available together with other structural feature predictors at: http://promoter.ics.uci.edu/BRNN-PRED/. PMID- 11933071 TI - Modifications to canonical structure sequence patterns: analysis for L1 and L3. AB - The conformation of five of the six hypervariable loops that form the antigen binding site of antibodies is limited to a small set of structures designated as canonical structures. The canonical structure model has been constituted as a fundamental tool for the modeling of antibodies. The detailed study of tens of crystallographic structures of antibodies has shown the validity of this model in the great majority of cases. The robustness of the forecast capacity of this model depends fundamentally on the precision with which the sequence patterns that characterize each canonical structure form can be defined. Nevertheless, due to the enormous quantity of structural information about antibodies generated during the last decade, it is difficult to avoid mistakes or confusion in the model. In the present work, we propose some corrections to the model for loops L1 and L3 that permit defining sequence patterns that avoid confusion and make better forecasting of the canonical structure model possible. PMID- 11933072 TI - Mutations in conserved regions 1, 2, and 3 of Raf-1 that activate transforming activity. AB - To investigate the role of Raf-1 in v-Ha-ras transformation, we have isolated and characterized a number of Raf-1 mutants that display increased transforming activity in Rat2 fibroblasts. A dipeptide deletion (Delta144-145) in the cysteine rich domain (CRD) of conserved region (CR) 1 increased the interaction between Raf-1 and v-Ha-ras effector loop mutants in the yeast two-hybrid system, supporting the proposal that the CRD serves as a secondary ras-binding domain. Many activating mutations were located in CR2. Two representative CR2 mutants (Delta250-258 and S257L) displayed increased interaction with v-Ha-ras effector loop mutants and with mitogen-activated protein kinase/extracellular signal regulated kinase (ERK) kinase (MEK) 1 in the two-hybrid system. One novel mutation in CR3 was recovered; G361S affected the third glycine of the GXGXXG protein kinase motif involved in ATP binding. Expression of G361S Raf-1 in Rat2 fibroblasts activated MEK and ERK. The CR1, CR2, and CR3 activating mutations, when combined in cis, cooperated in transforming Rat2 fibroblasts. Conversely, Raf-1 transforming activity was decreased when the S257L or G361S mutation was combined in cis with the R89E substitution, which disrupts ras-Raf interaction. This mutant analysis provides additional information about the distinct functions of individual Raf-1 regions and documents a novel genetic mechanism for activating an oncogenic kinase. PMID- 11933073 TI - Quinoxaline 1,4-dioxides: hypoxia-selective therapeutic agents. AB - A problem that confronts clinicians in the treatment of cancer is the resistance of hypoxic tumors to chemotherapy and radiation therapy. Thus, the development of new drugs that are toxic to hypoxic cells found in solid tumors is an important objective for effective anticancer chemotherapy. We recently showed that the heterocyclic aromatic N-oxides, quinoxaline 1,4-dioxides (QdNOs), are cytotoxic to tumor cells cultured under hypoxia. In this study, we evaluated the hypoxia selective toxicity of four diversely substituted QdNOs and determined their effect on the expression of hypoxia inducible factor (HIF) 1alpha in the human colon cancer cell line T-84. The various QdNOs were found to possess a 50- to 100 fold greater cytotoxicity to T-84 cells cultured under hypoxia compared with oxia. Interestingly, the hypoxia cytotoxicity ratio (HCR), the ratio of equitoxic concentrations of the drug under aerobic/anoxic conditions, was highly structure related and depended on the nature of the substituents on the QdNO heterocycle. The most cytotoxic 2-benzoyl-3-phenyl-6,7-dichloro derivative of QdNO (DCQ) was potent at a dose of 1 microM with an HCR of 100 and significantly reduced the levels of HIF-1alpha transcript and protein. The 2-benzoyl-3-phenyl derivative (BPQ) had a hypoxia potency of 20 microM and an HCR of 40. By contrast, the 2 aceto-3-methyl and the 2,3-tetramethylene (TMQ) derivatives of QdNO were much less cytotoxic under hypoxia (HCRs of 8.5 and 6.5, respectively) and reduced the expression of HIF-1alpha mRNA to a much lesser extent. Because the nonchlorinated analogue BPQ did not demonstrate behavior similar to that of DCQ, we hypothesize that the C-6, C-7-chlorine of DCQ might play a significant role in the selective hypoxic cytotoxicity of the drug. PMID- 11933074 TI - v-Ha-ras mitogenic signaling through superoxide and derived reactive oxygen species. AB - The ras proto-oncogene is frequently mutated in human tumors and functions to constitutively stimulate signal transduction cascades, resulting in unchecked proliferation and malignant transformation. In certain cells, superoxide functions as a signal-transduction messenger, mediating the downstream effects of ras and rac. We demonstrated previously that v-Ha-ras-transfected rat kidney epithelial cells (RECs) overproduced superoxide anion and that this superoxide production was mediated by ras. In the present study, we further demonstrated that v-Ha-ras overexpression transformed immortal nonmalignant RECs into malignant cancer cells; v-Ha-ras-transfected cells formed clones in soft agar, had high plating efficiency, and formed tumors in nude mice. Our data suggest that superoxide radical plays a role in ras-induced transformation; modulation of intracellular superoxide level by overexpression of manganese-containing superoxide dismutase or copper- and zinc-containing superoxide dismutase inhibited ras-induced transformation, as evidenced by in vitro studies of plating efficiency and by in vivo studies of tumor formation in nude mice. Overexpression of catalase (CAT) alone was found to have little effect on tumor cell growth, but overexpression of glutathione peroxidase 1 (GPx1) completely suppressed tumor cell growth in nude mice. This finding suggests that peroxides removed by GPx1, but not by CAT, are also involved in ras-induced transformation. PMID- 11933075 TI - Deregulated expression of the PCPH proto-oncogene in rat mammary tumors induced with 7,12-dimethylbenz[a]anthracene. AB - The PCPH proto-oncogene was identified by its frequent activation in Syrian hamster fetal cells exposed to 3-methylcholanthrene. We previously isolated human PCPH cDNA and studied its expression in normal human tissues. We report herein the pattern of PCPH expression in normal rat tissues. Each tissue expressed one major PCPH polypeptide that varied in molecular mass in different tissues. Normal mammary gland expressed a single PCPH polypeptide of 27 kDa. This PCPH form also was expressed in lactating mammary glands but at significantly greater levels. These results suggest the existence of tissue-specific regulatory mechanisms for PCPH expression that may be influenced by the differentiation stage. Our previous studies on the involvement of PCPH in human cancer showed that human breast tumor cell lines have frequent alterations in PCPH, including multiple PCPH polypeptide forms that are not expressed in normal cells. These cell lines also have frequent loss of a 27-kDa form identified as the only PCPH polypeptide expressed by normal human breast epithelial cells. In this study, we found that these same alterations occurred in vivo during mammary carcinogenesis in Sprague-Dawley rats treated with 7,12-dimethylbenz[a]anthracene, in both benign and malignant tumors, indicating that stable changes in PCPH expression took place early in the neoplastic process. Results showed that this experimental system is relevant to human breast carcinogenesis and provides an excellent model to study the molecular basis of the regulation of PCPH expression during normal differentiation and pathologic stages of neoplasia of the mammary gland and to analyze the role of PCPH in the carcinogenic process. Furthermore, the detection of atypical PCPH polypeptides in tumors suggests that PCPH immunodetection may be applied as a diagnostic tool for the early identification of neoplastic breast epithelial cells. PMID- 11933076 TI - Role of extracellular signal-regulated kinase pathway in RRR-alpha-tocopheryl succinate-induced differentiation of human MDA-MB-435 breast cancer cells. AB - RRR-alpha-tocopheryl succinate (vitamin E succinate, VES) induces differentiation of human breast cancer cells. Previous studies ruled out transforming growth factor-beta and c-jun N-terminal kinase involvement in VES-induced differentiation but implicated extracellular signal-regulated kinases (ERKs). Here we show that dominant-negative mutants of either mitogen-activated protein kinase kinase (MEK) 1 or ERK1 blocked VES-induced differentiation of MDA-MB-435 cells, as measured by induction of cytokeratin 18 and p21 (Waf1/Cip1) proteins. Blockage of c-jun protein expression using c-jun antisense oligonucleotides or expression of an inducible dominant-negative c-jun mutant protein inhibited VES induced differentiation. Elevated expression of wild-type c-jun alone was sufficient to induce cellular differentiation. A role for p21 (Waf1/Cip1) is implicated, in that p21 antisense oligomers blocked VES-induced differentiation. In summary, MEK1, ERK1, the transcription factor c-jun, and the cyclin-dependent kinase inhibitor p21 (Waf1/Cip1) play a part in VES-induced differentiation of human MDA-MB-435 breast cancer cells. PMID- 11933077 TI - Regulation of transcription of the intracellular interleukin-1 receptor antagonist gene by AP-1 in mouse carcinoma cells. AB - Interleukin-1 receptor antagonist (IL-1Ra) is involved in many processes, including epidermal inflammation and hyperplasia after irritation or injury. However, the mechanism by which intracellular IL-1Ra (icIL-1Ra) expression is regulated in mouse keratinocytes has not been reported. We found that the CH72 mouse carcinoma cell line constitutively expresses the icIL-1Ra mRNA. To study the transcriptional factors responsible for the constitutive expression of icIL 1Ra, we functionally characterized 4.5 kb of the 5' flanking region of the human icIL-1Ra gene in these cells. We first demonstrated that icIL-1Ra expression in these cells was regulated at the level of transcription. Deletion analysis of the promoter showed that regulatory elements for constitutive expression were located -158 to -49 bp upstream of the transcription start site for icIL-1Ra. We investigated the cis- and trans-acting factors required for icIL-1Ra expression. An activating protein-1 (AP-1) site was identified as the positive regulatory element necessary for the constitutive expression of the icIL-1Ra promoter in CH72 cells. Moreover, electrophoretic mobility shift assay and cotransfection experiments showed that c-jun and c-fos proteins bound to the AP-1 site and functionally transactivated the icIL-1Ra promoter in mouse carcinoma CH72 cells. PMID- 11933084 TI - Dendritic ruthenium porphyrins: a new class of highly selective catalysts for alkene epoxidation and cyclopropanation. AB - Attachment of Frechet-type poly(benzyl ether) dendrons [G-n] to carbonylruthenium(II) meso-tetraphenylporphyrin (5) using covalent etheric bonds forms a series of dendritic ruthenium(II) porphyrins 5-[G-n](m) (m=4, n=1, 2; m=8, n=0-2). The attachment was realized by treating the carbonylruthenium(II) complex of 5,10,15,20- tetrakis(4'-hydroxyphenyl)porphyrin or 5,10,15,20 tetrakis(3',5'-dihydroxyphenyl)porphyrin with [G-n]OSO(2)Me in refluxing dry acetone in the presence of potassium carbonate and [18]crown-6. Complexes 5-[G n](m) were characterized by UV/Vis, IR, and NMR spectroscopy and mass spectrometry. All of the dendritic ruthenium porphyrins are highly selective catalysts for epoxidation of alkenes with 2,6-dichloropyridine N-oxide (Cl(2)pyNO). The chemo- or diastereoselectivity increases with the generation number of the dendron and the number of dendrons attached to 5, and complex 5-[G 2](8) exhibits remarkable selectivity or turnover number in catalyzing the Cl(2)pyNO epoxidation of a variety of alkene substrates including styrene, trans /cis-stilbene, 2,2-dimethylchromene, cyclooctene, and unsaturated steroids such as cholesteryl esters and estratetraene derivative. The cyclopropanation of styrene and its para-substituted derivatives with ethyl diazoacetate catalyzed by 5-[G-2](8) is highly trans selective. PMID- 11933078 TI - Involvement of c-jun NH(2)-terminal kinases in resveratrol-induced activation of p53 and apoptosis. AB - Resveratrol, a constituent of grapes and other foods, is one of the most promising agents for cancer prevention. In a previous study, we showed that the antitumor activity of resveratrol occurs through extracellular signal-regulated protein kinases (ERKs) and p38 kinase-mediated p53 activation. In this study, we also determined that c-jun NH(2)-terminal kinases (JNKs) are involved in resveratrol-induced p53 activation and induction of apoptosis. In the JB6 mouse epidermal cell line, resveratrol activated JNKs dose-dependently within a dose range of 10-40 microM, the same dosage responsible for the inhibition of tumor promoter-induced cell transformation. Stable expression of a dominant negative mutant of JNK1 or disruption of the Jnk1 or Jnk2 gene markedly inhibited resveratrol-induced p53-dependent transcription activity and induction of apoptosis. Furthermore, resveratrol-activated JNKs were shown to phosphorylate p53 in vitro, but this activity was repressed in the cells expressing a dominant negative mutant of JNK1 or in Jnk1 or Jnk2 knockout (Jnk1(-/-) or Jnk2(-/-)) cells. These data suggested that JNKs act as mediators of resveratrol-induced activation of p53 and apoptosis, which may occur partially through p53 phosphorylation. PMID- 11933086 TI - Donor-acceptor oligoenes with a locked all-trans conformation: synthesis and linear and nonlinear optical properties. AB - A general synthetic approach to variously polarised merocyanines and a cyanine with enhanced thermal and (photo)chemical stability by a locked all-trans conformation (derived from a rigidified hexatriene unit and a variety of common donor and acceptor groups) is presented as well as a systematic study of their (non)linear optical properties. Apart from the UV/Vis absorption and fluorescence behaviour, the ground- and excited-state dipoles, the first-, second- and third order molecular polarisabilities were determined by electro-optical absorption measurements (EOAM) and degenerate four-wave mixing (DFWM) techniques in solution. Large values for the second- and third-order polarisability up to beta(0)=461x10(-50) C V(-2) m(3) (1242x10(-30) esu) and [gamma(LL)]=183x10(-60) C V(-3) m(4) (15x10(-34) esu) were found. The linear and nonlinear optical properties were related to the ground-state polarisation and the resonance structure of the chromophores. In order to reveal the influence of the length of the polymethinic chain (number of pi electrons within the chromophore), some lower homologues shortened by one C=C (double) bond were also taken into account. The unexpectedly high gamma values of some of the merocyanines cannot be explained by a two-level model. Molecular vibrational third- order polarisabilities (calculated from absolute Raman intensities in solution) were qualitatively correlated to the DFWM results. Furthermore, the dependence of the (13)C NMR chemical shifts of the polymethinic carbons within the merocyanines upon ground-state polarisation was investigated and compared to those within a corresponding cyanine. PMID- 11933087 TI - Cyclopentadienylrhodium coordination to alkenylarenes. AB - Photochemical reaction of [Rh(eta-C(5)H(5))(C(2)H(4))(2)] (5) with alkenyl benzene derivatives PhC(R(1))=CHR(2) results in the formation of four types of cyclopentadienylrhodium complexes: the mononuclear ethylene eta(2)-alkenylbenzene complexes [Rh(eta-C(5)H(5))(eta-C(2)H(4))(eta(2)-PhC(R(1))=CHR(2))] 9 a (R(1)=H, R(2)=Ph), 9 b (R(1)=Ph, R(2)=H), 9 c (R(1)=CH(3), R(2)=H), the mononuclear eta(4) alkenylbenzene complex [Rh(eta-C(5)H(5))[beta,alpha,1,2-eta-C(6)H(5)C(Ph)=CH(2)]] (10), the dinuclear mu-eta(4):eta(4)-alkenylbenzene complex [anti-[Rh(eta C(5)H(5))](2)[mu-beta,alpha,1,2-eta:3,4,5,6-eta-C(6)H(5)C(Ph)C=CH(2)]] (11), and the dinuclear rhodaindenyl complexes [Rh(eta-C(5)H(5))[1-3,8,9-eta-[1-(eta C(5)H(5))]-3-R(1)-1-rhodaindenyl]] 12 a (R(1)=Ph), 12 b (R(1)=CH(3)). Reaction of 5 with triisopropenylbenzene gives the dinuclear complex [[Rh(eta C(5)H(5))](2)(mu-beta,alpha,1,2-eta:beta',alpha',4,3-eta C(6)H(3)[C(CH(3))=CH(2)](3))] (13). In the complexes 9, only the olefinic side chain of the alkenylbenzene binds to the metal. In the complexes 10, 11, 12, and 13, an arene nucleus coordinates to rhodium as a 1,3-diene moiety (or part thereof). The rhodaindenyl complexes 12 result from C-H activation of the alkenylbenzene at the beta and ortho positions. The crystal and molecular structures of 9 a, 9 b, 10, 11, and 12 a, b were determined. The role of 9-11 and 13 as models for intermediates during alkenylbenzene-assisted self-assembly of tricobalt clusters is discussed. PMID- 11933085 TI - Metalloporphyrin-mediated asymmetric nitrogen-atom transfer to hydrocarbons: aziridination of alkenes and amidation of saturated C-H bonds catalyzed by chiral ruthenium and manganese porphyrins. AB - Chiral metalloporphyrins [Mn(Por*)(OH)(MeOH)] (1) and [Ru(Por*)(CO)(EtOH)] (2) catalyze asymmetric aziridination of aromatic alkenes and asymmetric amidation of benzylic hydrocarbons to give moderate enantiomeric excesses. The mass balance in these nitrogen-atom-transfer processes has been examined. With PhI=NTs as the nitrogen source, the aziridination of styrenes, trans-stilbene, 2 vinylnaphthalene, indene, and 2,2-dimethylchromene catalyzed by complex 1 or 2 resulted in up to 99 % substrate conversions and up to 94 % aziridine selectivities, whereas the amidation of ethylbenzenes, indan, tetralin, 1-, and 2 ethylnaphthalene catalyzed by complex 2 led to substrate conversions of up to 32 % and amide selectivities of up to 91 %. Complex 1 or 2 can also catalyze the asymmetric amidation of 4-methoxyethylbenzene, tetralin, and 2-ethylnaphthalene with "PhI(OAc)(2) + NH(2)SO(2)Me", affording the N-substituted methanesulfonamides in up to 56 % ee with substrate conversions of up to 34 % and amide selectivities of up to 92 %. Extension of the "complex 1 + PhI=NTs" or "complex 1 + PhI(OAc)(2) + NH(2)R (R=Ts, Ns)" amidation protocol to a steroid resulted in diastereoselective amidation of cholesteryl acetate at the allylic C H bonds at C-7 with substrate conversions of up to 49 % and amide selectivities of up to 90 % (alpha:beta ratio: up to 4.2:1). An aziridination- and amidation active chiral bis(tosylimido)ruthenium(VI) porphyrin, [Ru(Por*)(NTs)(2)] (3), and a ruthenium porphyrin aziridine adduct, [Ru(Por*)(CO)(TsAz)] (4, TsAz=N-tosyl-2- (4-chlorophenyl)aziridine), have been isolated from the reaction of 2 with PhI=NTs and N-tosyl-2-(4-chlorophenyl)aziridine, respectively. The imidoruthenium porphyrin 3 could be an active species in the aziridination or amidation catalyzed by complex 2 described above. The second-order rate constants for the reactions of 3 with styrenes, 2-vinylnaphthalene, indene, ethylbenzenes, and 2 ethylnaphthalene range from 3.7-42.5x10(-3) dm(3) mol(-1) s(-1). An X-ray structure determination of complex 4 reveals an O- rather than N-coordination of the aziridine axial ligand. The fact that the N-tosylaziridine in 4 does not adopt an N-coordination mode disfavors a concerted pathway in the aziridination by a tosylimido ruthenium porphyrin active species. PMID- 11933088 TI - Axial and equatorial hydrogen bonds: jet-cooled rotational spectrum of the pentamethylene sulfide...hydrogen fluoride complex. AB - Two different axial and equatorial hydrogen-bonded conformers of the complex formed by pentamethylene sulfide and hydrogen fluoride have been generated in a pulsed supersonic expansion and characterised by means of Fourier transform microwave spectroscopy. The ground-state rotational spectra of six isotopomers (C(5)H(10)S...HF, C(5)H(10)S ...DF, C(5)H(10)(34)S ...HF, (13)C(alpha)C(4)H(10)S ...HF, (13)C(beta)C(4)H(10)S...HF and (13)C(gamma)C(4)H(10)S ...HF) have been analysed for both conformers in the frequency range 5.5-18.5 GHz. The rotational parameters were used to derive C(s) structures for the conformers, with hydrogen fluoride pointing to the domain of the nonbonding electron pairs at either the axial or equatorial position of the sulfur atom. The axial form was found to be the more stable, in contrast with the observation for the pentamethylene sulfide...HCl complex. No equatorial-to-axial relaxation was observed when He or Ar were used as the carrier gas. The conformational behaviour is compared with that of related six-membered rings and discussed in terms of the existence of secondary hydrogen bonding between the halogen atom and the nearest H atoms of the methylene groups of the ring. No significant structural distortion of pentamethylene sulfide upon complexation was detected from a comparison with the structure of the isolated monomer. Finally, an ab initio study was carried out to complement the experimental results. PMID- 11933089 TI - Hydrothermal synthesis and characterization of the new layered fluorogallophosphate mu-23. AB - Mu-23, [(C(6)H(15)N(2))(C(6)H(16)N(2))Ga(5)F(6)(H(2)O)(2)(PO(4))(4)] x 4 H(2)O, the first layered fluorinated gallophosphate with a Ga/P molar ratio of 5:4, was obtained in the presence of fluoride ions with 1,4-dimethylpiperazine as an organic template. It crystallizes in the triclinic space group P1 (no. 2) with unit cell parameters a=8.735(11), b=8.864(5), c=12.636(10) A, alpha=98.36(5), beta=100.18(8), gamma=115.84(7) degrees. The layers consist of GaO(2)F(3)(H(2)O), GaO(4)F(2) octahedra, and GaO(4) and PO(4) tetrahedra; these moieties share their oxygen and some of their fluorine atoms. The connectivity scheme of these different polyhedra leads to the formation of eight-membered rings. PMID- 11933090 TI - The total synthesis of the annonaceous acetogenin, muricatetrocin C. AB - The total synthesis of the potential antitumour agent muricatetrocin C has provided an ideal stage for the exploitation and development of new chemistry. A convergent synthetic strategy has been realised incorporating three distinct pieces of methodology, these include a highly diastereoselective hetero-Diels Alder reaction to construct the butenolide terminus, an oxygen to carbon rearrangement to install the trans-2,5-disubstituted tetrahydrofuran ring and a spatial desymmetrisation process to afford the anti-diol unit. PMID- 11933091 TI - Synthesis and reactivity of an alpha-stannyl beta-silyl enamide. AB - We describe here an unprecedented approach to the synthesis of stereodefined 1,2 bis-substituted enamides. Starting from bis-metallated enamides, a palladium copper coupling reaction allows the introduction of various groups at C1, whilst iododesilylation affords a new entry for the functionalization at C2. PMID- 11933092 TI - Structure and electrochemistry of self-organized fullerene-lipid bilayer films. AB - The unique properties of C(60)-bearing artificial lipids with three C(16) (lipid 1), C(14) (lipid 2), or C(12) (lipid 3) alkyl chains have been characterized by a variety of techniques, including (13)C NMR, UV/Vis, and FT-IR spectroscopies, differential scanning calorimetry (DSC), X-ray diffraction, and electrochemistry. The (13)C NMR and UV/Vis spectra show that the lipids 1-3 have a closed aziridine structure at a 6/6-ring junction of C(60). The DSC data reveal that cast films of 1 exhibit two endothermic peaks at temperature ranges of 35-40 degrees C (main transition) and 47-49 degrees C (subtransition) in air, water, and 0.5 M aqueous tetraethylammonium chloride solution, while cast films of 2 and 3 each display one endothermic peak at 50-57 degrees C. The results of temperature-dependent FT IR and UV/Vis studies of cast films of 1-3, together with the above data, reveal that the main peak in the DSC thermogram of a film of 1 can be attributed to a typical phase transition as seen in lipid bilayer membranes, while the sub endothermic peak seen with 1 and the peaks for 2 and 3 stem from a change in the orientation of the C(60) moieties. X-ray diffraction patterns of each of the cast films of 1-3 show a diffraction peak corresponding to the (001) plane, suggesting the formation of molecular bilayer membrane structures. Cyclic voltammograms and Osteryoung square-wave voltammograms obtained from cast films of 1-3 on basal plane pyrolytic graphite (BPG) electrodes show strong temperature dependences. Finally, the thermodynamics of the binding of nine different alkylammonium ions and two alkylphosphonium ions to the electrogenerated radical monoanions and dianions of 1-3 cast on electrodes is described. PMID- 11933093 TI - Fluxional processes in diamagnetic and paramagnetic allyl dicarbonyl and 2 methylallyl dicarbonyl molybdenum histidinato complexes as revealed by spectroscopic data and density functional calculations. AB - This work describes a detailed study on the structure and dynamics of pseudooctahedral low-valent complexes of the type [Mo(His-N(epsilon)-R)(eta-2-R' allyl)(CO)(2)] (His=N(delta),N,O-L-histidinate; R=H, R'=H (1); R=C(2)H(4)CO(2)Me, R'=H (2); R=H, R'=Me (3); R=C(2)H(4)CO(2)Me, R'=Me (4)). These diamagnetic 18 electron complexes were comprehensively characterized spectroscopically and by X ray crystallography. In the solid state, the (substituted) allyl ligand is in an endo position in all compounds, but it is trans to the His-N(delta) atom in 1 and 2, whereas it is trans to the carboxylate O atom for the 2-Me-allyl compounds 3 and 4. In solution, both isomers are present in a solvent-dependent equilibrium. The third isomer (allyl trans to His-NH(2)) is not spectroscopically observed in solution. This is in agreement with the results from density functional (DFT) computations (BPW 91 functional) for 1 and 3, which predict a considerably higher energy (+6.3 and +5.9 kJ mol(-1), respectively) for this isomer. A likely path for isomerization is calculated, which is consistent with the activation energy determined by variable temperature NMR measurements. At least for 3, the preferred path involves several intermediates and a rotation of the 2-Me-allyl ligand. For the paramagnetic 17-electron congeners, DFT predicts the exo isomer of 3(+) with the 2-Me-allyl ligand trans to the carboxylate O atom to be by far the most stable isomer. For 1(+), an endo-exo equilibrium between the isomers with the allyl ligand trans to the carboxylate O atom is suggested. These suggestions are confirmed by EPR spectroscopy on the electrochemically generated species, which show signals for one- (4) and two- (2) metal-containing compounds. The appearance of the EPR spectra may be rationalized by inspection of the SOMOs from DFT calculations of the species in question. The notion of a metal-centered oxidation is also substantiated by IR spectroelectrochemistry and by UV/Vis spectra of the 17-electron complexes. Upon depleting the metal of electron density, the stretching vibrations of the carbonyl ligands shift more than 100 cm(-1) to higher wavenumbers, and the carbonyl vibration of the metal-coordinated carboxylate shifts by about 50 cm(-1). A color change from yellow to green upon oxidation is observed visually and quantified by the appearance of a new band at 622 nm (2(+)) and 546 nm (4(+)), respectively. PMID- 11933095 TI - Total synthesis of a stereoisomer of bistramide C and assignment of configuration of the natural product. AB - After the isolation of the bioactive polyether bistramide C from the marine ascidian Lissoclinum bistratum in 1988, NMR spectroscopic investigations over the next 12 years reduced the total number of possible stereoisomers of this compound from 1024 to 32. Based on the preparation of segments of the natural product as well as the total synthesis of a randomly selected stereoisomer of bistramide C, the stereochemical puzzle could be further simplified to eight possible stereoisomers. A convergent three-segment coupling strategy, the use of a common, D-glucose-derived intermediate for the preparation of pyran rings in two segments, a stereoselective photo-spiroketalization, and the use of azides to minimize protective group manipulations before segment couplings are highlights of the synthetic approach. The total synthesis also provided the key segments for a chiroptical analysis according to van't Hoff's principle of optical superposition, which was crucial for the assignment of a sole relative and absolute configuration of the natural product. Bistramide C represents therefore the first member of this class of structurally unusual marine polyethers whose configuration is known as a result of the combined use of synthetic and chiroptical tools. PMID- 11933094 TI - Evidence of complete hydrophobic coating of bombesin by trifluoroethanol in aqueous solution: an NMR spectroscopic and molecular dynamics study. AB - Bombesin is a tetradecapeptide that possesses a random coil structure in pure water. In the presence of 30 % (v/v) 2,2,2-trifluoroethanol (TFE), it adopts a partial helical conformation involving the C-terminal amino acids 6-14. This conformational change, known as the TFE effect, is studied here in terms of the solvation state of the peptide at different TFE concentrations by means of intermolecular homo- and heteronuclear NOE measurements. When an aqueous solution of bombesin is titrated with TFE, a continual decrease in the water/peptide interactions and a concomitant increase in the TFE/peptide interactions is observed, and at 30 % (v/v) TFE no homonuclear NOEs between water and the peptide can be detected. The conformational transition of the bombesin molecule is thus accompanied by a complete surface covering with TFE. A parallel molecular dynamics (MD) study of the peptide in aqueous solution with the single-point charge (SPC) water model and in a 30 % (v/v) TFE/water mixture with a recently developed TFE model has also been performed. The 10 ns simulations were in agreement with the experimental data. The calculations indicate stabilisation of the alpha-helix in the H(2)O/TFE mixture, in contrast to the situation in pure water, and clustering of the TFE molecules around the peptide. PMID- 11933096 TI - 1,8-bis(tetramethylguanidino)naphthalene (TMGN): a new, superbasic and kinetically active "proton sponge". AB - 1,8-Bis(tetramethylguanidino)naphthalene (TMGN, 1) is a new, readily accessible, and stable "proton sponge" with an experimental pK(BH(+)) value of 25.1 in MeCN, which is nearly seven orders of magnitude higher in basicity than the classical proton sponge 1,8-bis(dimethylamino)-naphthalene (DMAN). Because of the sterically less crowded character of the proton-accepting sp(2)-nitrogen atoms, TMGN also has a higher kinetic basicity than DMAN, which is shown by time resolved proton self-exchange reactions. TMGN is more resistant to hydrolysis and is a weaker nucleophile towards the alkylating agent EtI in comparison to the commercially available guanidine 7-methyl-1,5,7-triazabicyclo[4.4.0]dec-5-ene (MTBD). Crystal structures of the free base, of the mono- and bisprotonated base were determined. The dynamic behavior of all three species in solution was investigated by variable-temperature (1)H NMR experiments. DeltaG (++) values obtained by spectra simulation reveal a concerted mechanism of rotation about the C-N bonds of the protonated forms of TMGN. PMID- 11933097 TI - The proton affinity of the superbase 1,8-bis(tetramethylguanidino)naphthalene (TMGN) and some related compounds: a theoretical study. AB - The spatial and electronic structure of the very strong neutral organic bases bis(tetramethylguanidino)naphthalene (TMGN), 4,5 bis(tetramethylguanidino)fluorene (TMGF) and some related compounds are explored by ab initio computational methods. Their affinity towards the proton is scrutinized both in the gas phase and in solution in acetonitrile. The protonation at the most basic center (the imine nitrogen) yields asymmetric and relatively strong intramolecular hydrogen bonds (IHB). It is found that the angular strain effect and steric repulsion practically vanish in TMGN which implies that its high absolute proton affinity (APA) has its origin in the inherent basicity of the guanidine fragment and a relatively strong IHB in [TMGN]H(+). The nonbonded repulsions in TMGF are higher than in TMGN, which in conjunction with a slightly stronger IHB in the corresponding conjugate acid makes it more basic: APA(TMGF)>APA(TMGN). An interesting new phenomenon is observed in both TMGN and TMGF: the proton triggers the resonance stabilization not only in the directly bonded guanidine moiety, but also in the other guanidine fragment which is more distant from the proton, albeit in a less pronounced manner. The latter feature is termed a partial protonation. This supports the hydrogen bonding and contributes to the IHB stabilization. Convincing evidence is presented that the solvent effect in acetonitrile is determined by two antagonistic factors: 1) the intrinsic (gas phase) proton affinity and 2) the size effect which is given by the ratio between the positive charge in molecular cation (conjugate acid) and the magnitude of the molecular surface. The resulting pK(a) values are given by an interplay of these factors. PMID- 11933098 TI - Intercalation of cyclic ethers into vanadyl phosphate. AB - Two cyclic ethers, tetrahydrofuran (THF) and tetrahydropyran (THP), were intercalated into vanadyl phosphate and characterized by X-ray powder diffraction, thermogravimetry, and IR and Raman spectroscopy. Both compounds contain one molecule of ether per formula unit of VOPO(4) and show high thermal stability in comparison with VOPO(4) intercalates with other organic guest molecules. Both ethers are anchored to the VOPO(4) host layers by their oxygen atoms, which are coordinated to the vanadium atoms of the host. The probable arrangement of the tetrahydropyran molecules in the host interlayer space is derived from molecular simulations by the Cerius(2) 4.5 program. PMID- 11933099 TI - X-ray diffraction and molecular simulation study of the crystalline and liquid states of succinic anhydride. AB - The crystal structure of succinic anhydride was studied at five temperatures between 100 K and the melting point by single-crystal X-ray diffraction. The temperature dependence of molecular libration tensors was determined. Intermolecular interactions, in particular through unusually close molecule molecule contacts, are discussed, with a detailed calculation of electrostatic energies. A method for the adaptation of existing crystal force fields to molecular dynamics has been developed; the adapted force field was used to study molecular motion and rotational diffusion with increasing temperature. Equilibration of the crystalline system becomes impossible at a temperature very close to the experimental melting temperature, where a sudden transition to the liquid state occurs, and a partial kinetic picture of the melting process is obtained. After validation of the force field against experimental crystal data, the state equation of the liquid was predicted. Enthalpies of sublimation, melting, and vaporization were calculated. The dynamics of a solution of succinic anhydride in a nonpolar solvent was simulated, for a discussion of the aggregation process leading to demixing and to crystal nucleation. PMID- 11933100 TI - Additions of allenyl/propargyl organometallic reagents to 4-oxoazetidine-2 carbaldehydes: novel palladium-catalyzed domino reactions in allenynes. AB - Metal-mediated carbonyl allenylation and propargylation of 4-oxoazetidine-2 carbaldehydes were investigated in aqueous environment. Different propargyl bromide and metal promoters showed varied regio- and stereoselectivities on product formation. In addition, an unprecedented one-pot stereoselective synthesis of beta-chlorinated allylic alcohols, which can also be considered as functionalized allylsilanes, has been developed, which involves tin(IV) chloride mediated reaction of propargyltrimethylsilane and 4-oxoazetidine-2-carbaldehydes. Some of the resulting coupling products were submitted to transition metal catalyzed reactions, such as the allenic Pauson-Khand and palladium-catalyzed reactions, leading to novel fused or bridged tricyclic beta-lactams. Remarkably, a novel domino process, namely the allene cyclization/intramolecular Heck reaction was found. A likely mechanism for the cascade reaction should involve an intramolecular cyclization on a (pi-allyl)palladium complex and a Heck-type reaction. PMID- 11933101 TI - Reactions of gem-dibromo compounds with trialkylmagnesate reagents to yield alkylated organomagnesium compounds. AB - The reaction of gem-dibromocyclopropanes 5 with nBu(3)MgLi affords butylated cyclopropylmagnesium species that can be trapped with various electrophiles. The reaction of dibromomethylsilanes 12 requires the addition of a catalytic amount of CuCN small middle dot2 LiCl for smooth migration of the alkyl groups. The resultant alpha-silylpentylmagnesium compounds 16 react with electrophiles, such as acyl chlorides or alpha,beta-unsaturated ketones to afford alpha- or gamma silyl ketones, respectively. Treatment of dibromodisilylmethanes with Me(3)MgLi yields 1-bromo-1,1-disilylethanes 25 that can be converted into 1,1 disilylethenes 29 by dehydrobromination. PMID- 11933102 TI - Bonding and redox properties of [Os(3)(CO)(9)(tmbp)(L)] (tmbp=4,4',5,5' tetramethyl-2,2'-biphosphinine; L=CO, PPh(3)) clusters with an unprecedented electron-deficient metallic core and doubly bridging biphosphinine dianion. AB - Herein we describe in detail the bonding properties and electrochemical behavior of the first known triosmium carbonyl clusters with a coordinated redox-active ligand 4,4',5,5'-tetramethyl-2,2'-biphosphinine (tmbp), the phosphorus derivative of 2,2'-bipyridine. The clusters investigated were [Os(3)(CO)(10)(tmbp)] (1) and its derivative [Os(3)(CO)(9)(PPh(3))(tmbp)] (2). The crystal structures of both clusters are compared with those of relevant compounds; they served as the basis for density functional theory (DFT and time-dependent DFT) calculations. The experimental and theoretical data reveal an unexpected and unprecedented bridging coordination mode of tmbp, with each P atom bridging two metal atoms. The tmbp ligand is formally reduced by transfer of two electrons from the triangular cluster core that consequently lacks one of the metal-metal bonds. Both 1 and 2 therefore represent 50e(-) clusters with a coordinated 8e(-) donor, [tmbp](2-). The HOMO and LUMO of 1 and 2 possess a predominant contribution from different pi*(tmbp) orbitals, implying that the lowest energy excited state possesses a significant intraligand character. This is in agreement with the photostability of these clusters. DFT calculations also predict the experimentally observed structure of 1 to be the most stable one in a series of several plausible structural isomers. Stepwise two-electron electrochemical reduction of 1 and 2 results in dissociation of CO and PPh(3), respectively, and formation of the [Os(3)(CO)(9)(tmbp)](2-) ion. The initially produced radical anions of the parent clusters, in which the odd electron is predominantly localized on the tmbp ligand, are sufficiently stable at low temperatures and can be observed with IR spectroelectrochemistry. The electron-deficiency of the cluster core in 1 permits facile electrocatalytic substitution of a CO ligand by tertiary phosphane and phosphite donors. PMID- 11933104 TI - Analytical construct resins for analysis of solid-phase chemistry. AB - Rapid and unambiguous analysis of reactions performed on resin supports can be achieved by using "analytical constructs". These resins allow the synthesis of materials using solid-phase methods in the usual manner, but they also contain functionality enabling cleavage of analytically enhanced derivatives of the resin bound products. This is possible due to the use of two linkers bound in series to the polymer. Cleavage at the first linker yields the products attached to an analytical enhancer that facilitates detection. Orthogonal cleavage at the second linker yields the desired products in the usual manner. PMID- 11933105 TI - Novel applications of alkenyl zirconocenes. AB - Alkenyl zirconocene complexes are readily obtained by diverse processes including hydrozirconation, carbozirconation, and cyclozirconation of alkynes, transmetalations, and bond insertions. In combination with other metals, novel reaction manifolds can emerge and provide access to useful synthetic building blocks. Both catalytic asymmetric processes and highly diastereoselective multicomponent transformations that involve the formation of three or more new carbon-carbon bonds are feasible. This Concept paper summarizes the current state of the art and opportunities for future reaction discovery in this field of research, with particular emphasis on synergistic effects of bimetallic combinations of zirconocenes and zinc. PMID- 11933106 TI - Electronic properties of the silver-silver chloride cluster interface. AB - The objective of this study was to gain insight into the electronic structure of silver-silver chloride cluster composites and especially into the metal semiconductor interface. For this purpose a theoretical study of (AgCl)(n) (n=4, 32, 108, 192, and 256), of Ag(m) (m=1-9, 30, 115, 276, and 409), and of the cluster composites Ag(115)-(AgCl)(192) and Ag(409)-(AgCl)(192) has been carried out. Density of levels (DOL), local density of levels (l-DOL), and projection of surface states, as well as projection of properties of individual atoms or groups of atoms obtained in molecular orbital calculations, are shown to be powerful tools for gaining deep insight into the properties of these large systems. The Ag(115)-(AgCl)(192) aggregate, consisting of a cubic Ag(115) cluster without corner atoms on top of a cubic (AgCl)(192) cluster, was found to be remarkably stable with a cluster-to-cluster distance of about 280 pm, and a geometry in which the number of bonding interactions between the silver atoms of Ag(115) and the chloride ions of (AgCl)(192) is at its maximum. A sharp jump in charge distribution occurs at the Ag(115)-(AgCl)(192) composite interface. The first AgCl slab picks up negative charge from the two adjacent silver slabs, so that in total the silver cluster is positively charged. In addition, the core of the silver cluster is positively charged with respect to its outermost layer. The main reason for the charge transfer from the silver cluster to the silver chloride is the newly formed MIGS (metal induced gap states) in the energy-gap range of the silver chloride and the MIdS (metal induced d states) in the d orbital region. Their wave functions mix with orbitals of the silver cluster and with both the orbitals of the silver and the chloride ions of the silver chloride. The MIGS and the MIdS are of a quite localized nature. In them, nearest neighbor interactions dominate, with the exception of close-lying silver chloride surface states-which mix in to a large extent. We conclude that especially the MIGS not only influence the photochemical properties of silver chloride, but that their existence might be probed by appropriate spectroscopic measurements. PMID- 11933107 TI - A new efficient route for multigram asymmetric synthesis of alkannin and shikonin. AB - A short and convergent approach for the synthesis of alkannin, shikonin and shikalkin is presented. A Hauser-type annulation of cyanophthalide 26 with enone 7 affords the complete aromatic system in just one step with concomitant attachment of the entire side chain. Subsequent Corey's oxazaborolidine mediated asymmetric reduction of the above advanced intermediate, leads to the required isomer in high enantiomeric excess. Finally, a selective and high yielding deprotection protocol furnishes the title compounds as pure crystalline precipitates. Thus, a multigram synthesis of shikonin, alkannin and shikalkin is achieved in high yield and enantioselectivity. PMID- 11933108 TI - Croconic acid and alkali metal croconate salts: some new insights into an old story. AB - The solid-state structures of a series of alkali metal salts of the croconate dianion (C(5)O(5)(2-)) and of croconic acid (H(2)C(5)O(5)) have been determined. The alkali metal croconates were obtained by ring contraction of rhodizonic acid (H(2)C(6)O(6)), upon treatment with alkali metal hydroxides and recrystallisation from water. The novel species Na(2)C(5)O(5) x 2H(2)O, Rb(2)C(5)O(5) and Cs(2)C(5)O(5), as well as the mixed hydrogencroconate/croconate salt K(3)(HC(5)O(5))(C(5)O(5)) small middle dot2 H(2)O are described and compared with the Li(+), K(+) and NH(4)(+) salts. Single crystals of croconic acid were obtained by crystallisation of croconic acid in the presence of HCl. Crystal structure determinations showed that the C(5)O(5)(2-) ions tend to organize themselves in columns. The interplanar separations lie in the narrow range 3.12 3.42 A and do not necessarily reflect the presence of pi-stacking interactions. It is argued that the small interplanar separation is the result of a compromise between packing of flat croconate units and the spherical cations together with the water molecules that fill the coordination spheres of the alkali metal atoms. PMID- 11933110 TI - Ethnobiology of the Nilgiri hills, India. AB - The Nilgiri is a popular massif towering high in the Western Ghats in South India with an altitude of 2623 m. Nature has been magnanimous in bestowing Nilgiri district with rich evergreen temperate to tropical forests. A high degree of biodiversity, marked by varied flora and fauna of good therapeutic potential as well as the varied number of indigenous groups of people in this area, makes it very popular among herbalists. The district has six anthropologically well defined ethnic groups namely Todas, Kotas, Kurumbas, Irulas, Paniyas and Kattunayakas living here possibly since 1200 B.C. The present review highlights the ethnobiological profile of six indigenous populations and their dependence on ambient flora and fauna for traditional health care needs. It has been observed that about 2700 therapeutically potent plant species are available in this hill station of which almost all have come from local medicine. Some have been explored scientifically. However, about 150 plant species are still to be explored for their therapeutic potential. The ethnography, phytochemical and therapeutic uses as well as the anthropological perspectives of the local medicines have been discussed in this review. PMID- 11933111 TI - Induction of differentiation in rat C6 glioma cells with Saikosaponins. AB - The effects of saikosaponins (a, b(1), b(2), c, d), isolated from Bupleurum Radix, on the induction of differentiation in rat C6 glioma cells were studied. Saikosaponins a and d were shown to inhibit cell proliferation and alter cell morphology. In addition to cytostasis, the enzymatic activities of glutamine synthetase (GS) and 2',3'-cyclic nucleotide 3'-phosphohydrolase (CNP) were also noticeably increased after treatment with saikosaponin a. Nevertheless, saikosaponin d only showed an increase of GS activity, no significant changes in CNP activity were found. These results suggest that saikosaponin a can induce the differentiation of C6 glioma cells into astrocytes and/or oligodendrocytes, but saikosaponin d can only induce the differentiation of C6 glioma cells into astrocytes. PMID- 11933113 TI - Investigation into the toxicological effects of Catha edulis leaves: a short term study in animals. AB - In this study the short term (3 months) toxicological effects of varying levels of Catha edulis leaves were examined on the plasma concentration of liver enzymes and the histopathology of tissue sections of various organs including the liver, kidneys, spleen and testis. Both the biochemical and histopathological data demonstrated, initial signs of Catha edulis toxicity. Our results show a significant increase in plasma levels of alkaline phosphatase (ALP) and alanine aminotransferase (ALT) with all levels of Catha edulis leaves tested and throughout the treatment period. The increase of ALP was more prominent than that of ALT. The plasma levels of aspartate aminotransferase (AST) were only moderately increased at the higher dose (30%) in the later stages of treatment. In addition, a time-dependent gradual increase in indirect bilirubin with a concomitant decrease in direct bilirubin levels was observed with the 30% Catha edulis with no signs of haemolysis. The histopathology of tissue sections of the liver displayed evidence of congestion of the central liver veins as well as acute hepatocellular degenerative and regenerative activities in the tissue sections obtained from animals treated with both 20% and 30% Catha edulis. Similarly, histopathological examination of the tissue sections of the kidneys showed some lesions, and the degree of the lesion increased as the dose of Catha edulis leaves increased including: the presence of fat droplets particularly seen in the upper cortical tubules; acute cellular swelling; hyaline tubules; and acute tubular nephrosis. In contrast, Catha edulis treatment did not affect the spleen and increased the rate of spermatogenesis in male rabbits with the spermatozoa being quite evident, the Leydig cells were in good condition and were not affected by the doses given. PMID- 11933112 TI - LDH profiles of male and female rats treated with Vepacide. AB - In the present study we investigated the effect of vepacide, a neem-based compound, on the biochemical target enzyme lactate dehydrogenase (LDH) in different tissues of male and female albino Wistar rats treated orally with 80, 160 and 320 mg/kg (low, medium and high doses, respectively) for a period of 90 days. Prolonged administration of vepacide caused a significant increase of LDH activity in serum and lung tissues and a decrease in liver and kidney in both male and female rats when measured after 45 and 90 days of daily treatment. Females were more susceptible than males with regard to serum and kidney LDH showing sexual dimorphism in the treated rats. Recovery was observed in the affected enzyme after 28 days post treatment (withdrawal study). A positive correlation was observed with regard to this enzyme between serum and lung tissues, whereas for serum versus liver and kidney there was a negative correlation. The effect of vepacide was more pronounced in the lung tissue followed by liver and kidney tissues. Necrosis of the liver and kidney tissues was observed but in the lung tissue an increase in the LDH enzyme was seen. Therefore, it was concluded that the increase in LDH could be indicative of a stress adaptive response to the toxicant. PMID- 11933114 TI - Synthesis of plumbagin derivatives and their inhibitory activities against Ehrlich ascites carcinoma in vivo and Leishmania donovani Promastigotes in vitro. AB - Plumbagin, a plant-derived bioactive naphthoquinonoid compound, was converted to a hydroquinonoid derivative, which was studied for its tumour-inhibitory and antileishmanial activities for the first time. A similar chemical transformation was undertaken on an analogous dimeric compound, diospyrin, and its bioassay results were compared with those of the plumbagin derivative. Synthesis of the derivative of plumbagin did not result in a marked enhancement of the tumour inhibitory activity, whereas the improvement was obvious in the case of diospyrin vis a vis its hydroquinonoid analogue. The conversion of diospyrin to the hydroquinonoid compound also led to a substantial increase in the antileishmanial activity, while a similar conversion of plumbagin failed to do so. PMID- 11933116 TI - The in vitro effect of Viscum album (VA) extract on DNA repair of peripheral blood mononuclear cells (PBMC) in cancer patients. AB - Viscum album (VA) extract as an immunomodulator was tested in an in vitro model to investigate DNA repair in damaged peripheral blood mononuclear cells (PBMC) of ten breast cancer patients. The cells were exposed by gamma rays or 4 hydroxycyclophosphamide (4-HCy). Two hours after exposure the following were measured, without or with VA extract (1) DNA repair using the alkaline sucrose gradient for the sedimentation of DNA strand breaks, (2) DNA-gamma-production in the supernatant of the cultured cells. The VA extract led to an improvement of DNA repair in gamma-ray or 4-HCy damaged PBMC and to a significant increase of the IFN-gamma-production both in undamaged and in damaged cells. The results indicate that the VA extract affects positively DNA repair in PBMC damaged by two different agents and suggest that an increased IFN-gamma-production may play an important role in the DNA repair process. PMID- 11933117 TI - Antioxidants from the bark of Burkea africana, an African medicinal plant. AB - The bark of the tree Burkea africana is used medicinally in large areas of sub Saharan Africa. The constituents responsible for its putative activity are not well known. We have investigated the bark of B. africana for antioxidant and radical scavenging activity. A hydroethanol bark extract showed high activity, and most of this activity was located in semipolar fractions of the extract. From chromatographic purification and spectroscopical structure studies, we conclude that the active constituents are proanthocyanidins. Two major components appear to be fisetinidol-(4alpha- --> 8)-catechin 3-gallate and bis-fisetinidol-(4alpha- --> 6, 4alpha- --> 8)-catechin 3-gallate. The latter compound is a new natural product. Smaller amounts of monomeric flavan-3-ols (catechin, epicatechin and fisetinidol) were also found. PMID- 11933115 TI - Polysaccharides isolated from Echinacea purpurea herba cell cultures to counteract undesired effects of chemotherapy--a pilot study. AB - In an open prospective study with matched historical controls we aimed to evaluate whether a polysaccharide fraction isolated from the herb Echinacea purpurea could counteract the undesired effects of chemotherapy. Fifteen patients with advanced gastric cancer undergoing palliative chemotherapy with etoposide, leucovorin and 5-fluorouracil (ELF) received for 10 days (beginning 3 days before chemotherapy) daily i.v. injections of 2 mg of a polysaccharide fraction isolated from Echinacea purpurea herb cell cultures (EPS-EPO VIIa). The median number of leukocytes 14-16 days after chemotherapy was 3630/microL (range 1470-5770) in the patients receiving EPS-EPO VIIa compared with 2370/microL (870-3950) in the patients of the historical control group (p = 0.015). EPS-EPO VIIa had no clinically relevant effects on phagocytic activity of granulocytes or on lymphocyte subpopulations. Sixty-eight adverse events including two deaths were observed, most likely due to chemotherapy and the general condition of the patients. However, an association with the test intervention cannot be ruled out completely. The results of this pilot study suggest that EPS-EPO VIIa might be effective in reducing chemotherapy-induced leukopenia. The efficacy and safety should be investigated in further studies. PMID- 11933118 TI - In vitro Inhibition of HHV-6 replication by sophocarpines. AB - The virostatic activity of sophocarpines and gancyclovir (GCV) was tested using HHV-6 Z29 strain and Molt-3 cells. The cytotoxic (IC(50)) and the antiviral (ED(50)) values were first experimentally determined and selective indices (SI) were then calculated. The SI values for sophocarpines 1 and 2 and GCV were in the order 184, 183, and 23, respectively. Though preliminary, these findings indicate that sophocarpines have the potential to inhibit HHV-6 replication. PMID- 11933119 TI - Effects of Rosa canina fruit extract on neutrophil respiratory burst. AB - Respiratory burst leads polymorphonuclear neutrophils (PMN) to produce reactive oxygen species (ROS) such as superoxide anions (O(2)(o-)), hypochlorous acid (HOCl) and hydrogen peroxide (H(2)O(2)) which may possess deleterious effects for the organism. Rosa canina fruits are well known to contain a large amount of vitamin C which is antioxidant. This study was focused on the polyphenolics contained in rose hips to evaluate their antioxidative properties. We prepared a rose hip extract deprived of vitamin C. The extract contained mainly phenolics such as proanthocyanidins and flavonoids. We investigated its effects directly against (O(2)(o-)), HOCl and H(2)O(2) and investigated its effects on isolated PMN. For that, in vitro inflammatory conditions were reproduced by stimulating PMN with stimuli having different transductional pathways, in order to determine a possible mechanism of action. The results showed that the extract can inhibit ROS tested in acellular and cellular systems. The IC(50) obtained were 5.73 mg/L, 1.33 mg/L and 2.34 mg/L respectively for (O(2)(o-)), HOCl and H(2)O(2) in acellular experiments. For cellular experiments, the IC(50) were quite similar. Thus, the extract did not present an effect on PMN metabolism. Therefore, the antioxidative effects of Rosa canina are due not only to vitamin C but also to polyphenolics. PMID- 11933120 TI - Immunomodulatory activity of septilin, a polyherbal preparation. AB - Septilin is a polyherbal preparation, claimed to be effective in conditions such as chronic stubborn URTI, tonsillitis, cutaneous infections, dental infections and also prescribed as a health supplement. In view of this, the present experimental study was undertaken to evaluate the effect of Septilin on different arms of the immune system. The experimental animals (male albino rats and mice) were divided into three groups. Group I received distilled water; group II received Septilin in a dose of 1 g/kg (rats) or 1.5 g/kg (mice); group III received Septilin 2 g/kg (rats) or 3 g/kg (mice) orally for 28 days. They were evaluated for immunological function on day 29 by studying weight gain, resistance against E. coli sepsis, haemogram, phagocytic activity of PMN cells and reticuloendothelial system, delayed hypersensitivity to oxazolone and the plaque forming cell response of splenic lymphocytes to sheep erythrocytes. Neither of the doses of Septilin altered weight gain, absolute lymphocyte counts, or host resistance against E. coli sepsis. The higher dose of Septilin reduced phagocytic activity of the PMN cells/reticuloendothelial system, but both doses increased the percentage and absolute number of circulating neutrophils, stimulated humoral immunity and suppressed cellular immunity. Thus, Septilin has dual effects on the immune system, with lower doses showing greater stimulant and higher doses showing predominantly suppressant effects. PMID- 11933121 TI - Effect of ganhuangenin obtained from Scutellaria radix on the chemical mediator production of peritoneal exudate cells and immunoglobulin E level of mesenteric lymph node lymphocytes in Sprague-Dawley rats. AB - We previously showed that ganhuangenin (GHG) has beneficial antioxidative properties against lipid peroxidation in tissues of Sprague-Dawley (SD) rats. In this study, the effect of GHG on immunoglobulin E (IgE), histamine and leukotriene B(4) (LTB(4)) levels of cells isolated from SD rats was examined with regard to the manifestation of the type I allergic reaction. We showed that GHG inhibited the IgE production of mesenteric lymph node (MLN) lymphocytes. In the presence or absence of concanavalin A (ConA), the concentration used did not exert a toxic effect against MLN lymphocytes. Interestingly, the increase in the IgE content and lipid peroxidation induced by ConA was alleviated in the presence of GHG. Moreover, GHG also inhibited histamine release from the peritoneal exudate cells stimulated with a calcium ionophore, A23187, or with the histamine releaser, compound 48/80. In the case of LTB(4), GHG markedly inhibited its release at a concentration of 100 microM. Thus, it is concluded that GHG may block the common pathway for the release of histamine and LTB(4), and that the IgE level is responsible for the lipid peroxidation induced by ConA. PMID- 11933122 TI - Effect of curcumin on ethanol-induced stress on mononuclear cells. AB - Blood cells in circulation are exposed to a wide variety of stress-causing agents, causing a number of changes including interactions with other cells and the extracellular matrix of the endothelial wall. In order to understand the role of curcumin, an antioxidant principle from Curcuma longa Linn., on blood mononuclear cells from rabbits given ethanol for 30 days and ethanol with curcumin, cells were isolated and an attachment assay was carried out. The monocytes from ethanol-treated rabbits showed a lesser attachment to collagen, the major component of the vessel wall subendothelium, and those from curcumin treated animals along with ethanol showed a higher affinity to collagen, causing an alteration in the attachment of monocyte to collagen due to ethanol-induced stress. PMID- 11933123 TI - Antibacterial activity of Harungana madagascariensis leaf extracts. AB - Aqueous extracts of the leaf of Harungana madagascariensis were analysed phytochemically and evaluated for antimicrobial activity against strains of Bacillus subtilis, Staphylococcus aureus, Escherichia coli, Salmonella typhi and Pseudomonas aeruginosa. Glycosides, tannins, saponins, flavonoids and alkaloids were detected in the plant material. B. subtilis, E. coli and S. typhi, but not Ps. aeruginosa, showed susceptibility at MICs of 2.0 and 15.6 mg/mL; and MBCs of 2.0-3.9 mg/mL and 15.6-31.3 mg/mL, respectively, for the cold and hot extracts. Staph. aureus showed susceptibility only to the hot extract. Concentrations of 2.5-10.0 mg/mL of the cold extract killed over 7 log(10) of the test bacterial population within 30-60 min of exposure. The hot extract needed higher concentrations and longer treatment to achieve similar levels of bacterial cell killing. The results provide a rationalization for the traditional use of H. madagascariensis leaf extracts for the treatment of gastrointestinal disorders. PMID- 11933124 TI - The effect of Polbax extract on lipofuscin accumulation in cultured neonatal rat cardiac myocytes. AB - Polbax, a water-soluble extract of fresh pollen grains and pistils, was tested for its ability to influence the accumulation of lipofuscin (age pigment) in cultured neonatal rat cardiac myocytes. Exposure for 3 weeks to Polbax at concentrations of 0.1, 1.0 or 10 mg/L decreased lipofuscin accumulation morphometrically assayed using laser scanning microscopy images (green excitation light) of formaldehyde-fixed cells, by 24%, 41% or 43%, respectively. Based on the knowledge that oxidative stress and iron-catalysed peroxidation play an important role in lipofuscinogenesis, we suggest that Polbax may slow lipofuscin formation due to antioxidant activities, perhaps involving intralysosomal dismutation of superoxide produced by autophagocytosed mitochondria and/or iron chelation. PMID- 11933125 TI - Chemical composition and biological activity of the volatiles of Anthemis melampodina and Pluchea dioscoridis. AB - The volatile fractions obtained by hydrodistillation of the fresh leaves of Anthemis melampodina and Pluchea dioscoridis were analysed by GC-MS technique. Of 38 components identified in the volatile oil of A. melampodina, santolinatriene was the major component (27.33%). The oil was characterized by a high percentage of monoterpene hydrocarbons (49.94%) while sesquiterpene hydrocarbons and oxygenated sesquiterpenes represented only 7.41% and 11.43% of the oil. 36 components were identified in the volatile oil of P. dioscoridis. Farnesol was the major component (16.50%) accompanied by a high percentage of sesquiterpene alcohols. Oxygenated sesquiterpenes (26.43%) and sesquiterpene hydrocarbons (39.43%) represented the main constituents in the oil. P. dioscoridis showed a marked mosquito larvicidal activity against Culex pipiens (LC(50) 71.86 ppm), while A. melampodina was moderately active (LC(50) 139.42 ppm). PMID- 11933126 TI - Screening of Chinese and Mongolian herbal drugs for anti-human immunodeficiency virus type 1 (HIV-1) activity. AB - Water and methanol extracts of 30 Chinese and Mongolian medicinal plants were tested for their human immunodeficiency virus type-1 (HIV-1) inhibitory activity. Of the 60 extracts, 23 showed anti-HIV activity. Bioassay-guided fractionation of one of the most active extracts, the methanol extract of the root tuber of Stephania cepharantha, led to the isolation of two alkaloids, aromoline and FK 3000 as potent inhibitory substances. They completely inhibited the cytopathic effects of HIV-1 on MT-4 cells at 31.3 and 7.8 microg/mL, respectively. PMID- 11933127 TI - Effect of Mimosa pudica root powder on oestrous cycle and ovulation in cycling female albino rat, Rattus norvegicus. AB - Mimosa pudica root powder (150 mg/kg body weight) when administered intragastrically, altered the oestrous cycle pattern in female Rattus norvegicus. Nucleated and cornified cells were absent in all rats. The smear was characterized by leucocytes only, as in dioestrus, which persisted for 2 weeks. There was a significant reduction in the number of normal ova in rats treated with the root powder compared with the control rats, and a significant increase in the number of degenerated ova. PMID- 11933128 TI - Antiulcerogenic and analgesic effects of the Austroplenckia populnea extracts in mice. AB - Austroplenckia populnea (Reiss.) Lund. is a Brazilian Cerrado plant belonging to the Celastraceae family. Hexane and methanol extracts of leaves were investigated for their antiulcerogenic (ethanol and indomethacin/bethanecol induced gastric damage) and analgesic (writhing and tail-flick tests) activities in mice. Acute toxic effects also were evaluated. Oral administration of both extracts at a dose of 1000 mg/kg significantly reduced the total area of the lesion, the relative area of lesion and the ulcerative index in ethanol-induced gastric damage, but both extracts were inactive in the indomethacin/bethanecol-induced gastric damage test. A dose dependent effect was determined with the hexane extract in the ethanol-induced lesions test. The hexane and methanol extracts reduced the number of contortions in the writhing test, but both extracts were inactive in the tail flick immersion test. PMID- 11933129 TI - Brine shrimp lethality assay of Bacopa monnieri. AB - Successive petroleum ether, chloroform, ethanol and water extracts, a saponin rich fraction (SRF) and bacoside A isolated from Bacopa monnieri were tested for brine shrimp lethality. Successive ethanol extracts and SRF showed potent activity. Bacoside A showed the maximum activity with a LC(50) of 38.3 microg/mL. The results confirmed the previous reports of an anticancer effect of Bacopa monnieri and suggest bacoside A as the active constituent. PMID- 11933130 TI - Comparative study of Venostasin and Pycnogenol in chronic venous insufficiency. AB - The aim of this study was to compare the efficacy of Venostasin (horse chestnut seed extract) and Pycnogenol (French maritime pine bark extract) in the treatment of chronic venous insufficiency (CVI). In an open, controlled comparative study 40 patients with diagnosed CVI were treated either with 600 mg chestnut seed extract per day or 360 mg Pycnogenol per day over a period of 4 weeks. The following parameters were investigated before the start of treatment and after 2 and 4 weeks of treatment: circumference of the lower legs and rating of subjective symptoms (scores) of pain, cramps, night-time swelling, feeling of "heaviness", and reddening of the skin. In addition, blood levels of cholesterol LDL and HDL were determined before and at the end of treatment. Pycnogenol significantly reduced the circumference of the lower limbs and significantly improved subjective symptoms. Furthermore, Pycnogenol significantly decreased cholesterol and LDL values in the blood, whereas HDL remained unaffected. Venostasin only moderately but not significantly, reduced the circumference of the lower limbs and marginally improved symptoms. Venostasin had no influence on the determined lipid values. Both medications were equally well tolerated. In conclusion, Pycnogenol was found to be more efficacious than Venostasin for the treatment of CVI. PMID- 11933131 TI - Vasodilating properties of extracts from the leaves of Musanga cecropioides (R. Brown). AB - The mechanisms of action involved in the hypotensive properties of the aqueous extract of the leaves of Musanga cecropioides were investigated. The effect of the aqueous leaf extract of M. cecropioides, found to contain mostly saponins, flavonoids and procyanidins, was investigated on vascular smooth muscle and also in an in vivo direct invasive blood pressure study in both normotensive and hypertensive rats. The hypotensive or antihypertensive properties of the extracts appear to be due partly to a direct or indirect vasodilator effect and also to some alpha(1)- and beta(2)-adrenergic blocking effects. The extract also exhibited significant endothelium-dependent vascular smooth muscle relaxation, accounted for by the release of nitric oxide (NO), and induced significant angiotensin converting enzyme (ACE) inhibitory effects thereby supporting its vasodilator mechanism of action. PMID- 11933132 TI - Pharmacological properties of the methanol extract from Mentha suaveolens Ehrh. AB - The present study analyses the pharmacological activity in in vivo and in vitro models of the methanol extract obtained from the leaves and steams of Menta suaveolens Ehrh. This extract lacked toxicity, but exhibited a central nervous system depressant action; an analgesic effect in models of chemical and mechanical stimulation suggesting the induction of a peripheral analgesic response. The extract also exhibited an antiinflammatory action inhibiting the rat paw oedema induced by carrageenin. Moreover, the in vitro studies showed a significant diminution in the contractile effects induced by histamine, serotonin and acetylcholine. PMID- 11933133 TI - Recuperative effect of Semecarpus anacardium linn. nut milk extract on carbohydrate metabolizing enzymes in experimental mammary carcinoma-bearing rats. AB - Semecarpus anacardium Linn. of the family Anacardiaceae has many applications in the Ayurvedic and Siddha systems of medicine. We have tested the antitumour activity of Semecarpus anacardium nut extract against experimental mammary carcinoma in animals. As there is a direct relationship between the proliferation of tumour cells and the activities of the glycolytic and gluconeogenic enzymes, we studied changes in the activities of enzymes involved in this metabolic pathway in the liver and kidney. The enzymes investigated were glycolytic enzymes, namely hexokinase, phosphoglucoisomerase, aldolase and the gluconeogenic enzymes, namely glucose-6-phosphatase and fructose-1,6-biphosphatase in experimental rats. A significant rise in glycolytic enzyme activities and a simultaneous fall in gluconeogenic enzyme activities were found in mammary carcinoma bearing rats. Drug administration returned these enzyme activities to their respective control activities. PMID- 11933134 TI - Effect of Perilla frutescens on nitric oxide production and DNA synthesis in cultured murine vascular smooth muscle cells. AB - The effects of perilla (Perilla frutescens, Labiatae) on murine cultured vascular smooth muscle cells (VSMC) were investigated. The water extract of perilla leaves induced nitric oxide (NO) production of VSMC and this effect was synergistically augmented when combined with interferon (IFN)-gamma or tumour necrosis factor (TNF)-alpha, while the perilla extract significantly inhibited NO production induced by IFN-gamma combined with lipopolysaccharide (LPS). Northern blot analysis revealed that these effects of the perilla extract paralleled mRNA expression of inducible nitric oxide synthase. However, the perilla extract significantly inhibited platelet derived growth factor (PDGF) or TNF-alpha induced VSMC proliferation measured as DNA synthesis. The inhibitory effect of the perilla extract on TNF-alpha-induced VSMC proliferation was significantly suppressed by N(G)-monomethyl-L-arginine, a non-specific nitric synthase inhibitor, suggesting that this effect was partially mediated by NO production as an autocrine/paracrine factor. The present findings suggest that perilla would be useful for the prevention of vascular diseases such as arteriosclerosis. PMID- 11933135 TI - Effects of extract from Angelica keiskei and its component, cynaroside, on the hepatic bromobenzene-metabolizing enzyme system in rats. AB - The effects of Angelica keiskei Koidz. on hepatic lipid peroxide and the activities of free radical generating and scavenging enzymes were investigated in bromobenzene-induced hepatic lipid peroxidation in rats. The level of lipid peroxide elevated by bromobenzene was significantly reduced by the methanol extract from the aerial parts of A. keiskei and its component, cynaroside. Epoxide hydrolase activity was decreased significantly by the treatment of bromobenzene. However, the enzyme activity was restored in the liver of rats given the methanol extract and cynaroside. The results suggest that the reduction of bromobenzene-induced hepatic lipid peroxidation by the extract of A. keiskei and cynaroside under our experimental conditions is thought to be through enhancing the activity of epoxide hydrolase, an enzyme removing bromobenzene epoxide. PMID- 11933136 TI - Effects of black tea extract on carbon tetrachloride-induced lipid peroxidation in liver, kidneys, and testes of rats. AB - Previous studies have shown that green tea and black tea have antioxidant effects and chemopreventive activity against chronic disease including some forms of cancer. We have, therefore, examined the effects of an aqueous extract of black tea against carbon tetrachloride-induced lipid peroxidation as determined by the formation of thiobarbituric acid reactive substances in liver, kidneys and testes of rats. A 0.7% black tea extract was used which contained 2 mg of black tea extract solids per mL. Black tea was administered as drinking water for 3, 6, 9 and 12 months before and during carbon tetrachloride (CCl(4)) treatment in female and male rats. Rats were treated with a single oral dose of CCl(4) 1.0 mL/kg. All rats were killed 24 h after CCl(4) treatment. All animals were dosed with CCl(4) at the end of the 3, 6, 9, and 12 month of treatment. Black tea treatment for 75 days produced a decrease in CCl(4)-induced hepatic lipid peroxidation but significant decreases in thiobarbituric acid reactive substances occurred 3 months after treatment in both female and male rats. In liver and kidneys, black tea alone increased lipid peroxidation by 30%-50% in female and male rats. However, black tea decreased CCl(4)-induced lipid peroxidation in liver of female and male rats by approximately 49% and 37%, respectively. Black tea decreased CCl(4)-induced lipid peroxidation in testes by approximately 37% at a dose of 1.0 mL CCl(4)/kg. These results suggest that the protective effects of black tea against CCl(4)-induced lipid peroxidation in liver, kidneys and testes is due at least partly to its antioxidant properties, scavenging CCl(4)-associated free radicals. PMID- 11933137 TI - Antioxidant properties of silybin glycosides. AB - New soluble derivatives of the hepatoprotective flavonolignan silybin (1), namely silybin galactoside (2), glucoside (3), lactoside (4) and maltoside (5) were investigated for their radical scavenging and antilipoperoxidation properties. According to cyclic voltammetry the results show that glycosides are weaker electron donors than silybin, although it was of interest that they were found to be more potent scavengers of the 1,1-diphenyl-2-picrylhydrazyl and the 2,2'-azino bis(3-ethylbenzothiazoline-6-sulphonic acid)-derived radicals. The glycosides (2) (5) were more efficient than silybin in preventing tert-butylhydroperoxide induced lipoperoxidation of rat liver mitochondrial membranes. Furthermore, glycosides (2)-(5) were significantly more cytoprotective than silybin in tert butylhydroperoxide-damaged rat erythrocytes and primary hepatocyte cultures. Glycosylation of silybin substantially reduced its toxic effects in primary cultured hepatocytes observed during prolonged incubation. These results suggest that silybin glycosides are suitable soluble derivatives of silybin for experimental studies and may have therapeutic potential. PMID- 11933138 TI - Inhibition of tumour growth and inflammation by consumption of tea. AB - The antitumour effect of tea was evaluated in the 3-methylcholanthrene (3-MC) induced solid tumour model in mice. Both black and green tea inhibited tumour growth and prevented metastasis. Histopathological study showed that tea treatment was able to reduce malignancy. Superoxide dismutase (SOD), a free radical scavenger, was found to be significantly increased in the serum of mice administered tea. Moreover, tea extracts were able to reduce the level of thiobarbituric acid reactive substance (TBARS) in the sera of mice. Tea extracts (both black and green) also showed antiinflammatory activity in the carrageenan induced paw oedema model in the rat. PMID- 11933140 TI - The in vitro antioxidative activity of some traditional Zulu medicinal plants. AB - Methanol extracts prepared separately from the roots, stems and leaves of four traditional Zulu medicinal plants (Rhoicissus digitata, R. rhomboidea, R. tomentosa and R. tridentata) were tested for their antioxidant activity. The extracts of R. rhomboidea and R. tridentata inhibited the activities of the 1, 1' diphenyl-2-picryhydrazyl free radical, xanthine oxidase, and also prevented production of thiobarbituric acid reactive substances and free radical mediated DNA sugar damage. The extracts had a strong chelating effect on Fe(2+) ions. R. digitata and R. tomentosa extracts, however, possessed some prooxidative properties at high concentrations. In view of these results, it is apparent that the antioxidative activity of these Zulu medicinal plants plays an important role in the healing of the various diseases that they are used for. PMID- 11933139 TI - Preventive effect of the Taiwan folk medicine Ixeris laevigata var. oldhami on alpha-naphthyl-isothiocyanate and carbon tetrachloride-induced acute liver injury in rats. AB - The hepatoprotective effects of Ixeris laevigata Sch-Bip. var. oldhami Kitam. (IL) were studied on cholestatic hepatitis induced by alpha naphthylisothiocyanate (ANIT, 100 mg/10 mL/kg, in olive oil, i.p.) and acute hepatitis induced by carbon tetrachloride (20% CCl(4)/olive oil, 1.5 mL/kg, i.p.) in rats. Hepatoprotective activity was monitored by estimating the serum transaminases levels and the histopathological changes in the livers of experimental rats. The pretreatment of animals with IL, extract (0.3-2.0 g/kg orally) significantly inhibited the acute elevation of serum transaminases, as well as the hepatotoxin-induced histopathological changes in the livers of the experimental rats. PMID- 11933141 TI - Inhibitory effects of quinones on RNase H activity associated with HIV-1 reverse transcriptase. AB - In an effort to develop new drugs preventing the growth of human immunodeficiency virus (HIV), we developed an in vitro assay method of ribonuclease H (RNase H) activity associated with reverse transcriptase (RT) from HIV-1. Some naphthoquinones, such as 1,4-naphthoquinone (1), vitamin K(3) (2), juglone (3) and plumbagin (6), moderately inhibited RNase H activity, and others, including naphthazarin (5) and shikonins (8-9, 18-23), showed weak inhibition. Diterpenoid quinones, tanshinones (24-28), had also moderate inhibition against RNase H activity. Of these quinones, compound 1 showed the most potent inhibition on RNase H activity with a 50% inhibitory concentration (IC(50)) of 9.5 microM, together with moderate inhibition against RNA-dependent and DNA-dependent DNA polymerase (RDDP and DDDP) activities with IC(50) values of 69 and 36 microM, respectively. Compounds 3 and 5 showed significant inhibition against RDDP (IC(50) = 8 and 10 microM, respectively) and DDDP (IC(50) = 5 and 7 microM, respectively) activities. The structure-activity relationship of the naphthoquinones suggested that non-hydroxylated naphthoquinones (1 and 2) showed significant inhibition of RNase H activity, whereas 5-hydroxylated naphthoquinones (3 and 5) showed potent inhibition against RDDP and DDDP activities. PMID- 11933143 TI - Insect growth inhibitor activity of arjunolic acid isolated from Cornus capitata. AB - Arjunolic acid (a pentacyclictrihydroxytriterpenic acid) isolated from the stem of Cornus capitata (Cornaceae) exhibited significant inhibitory activity towards 4th instar larvae of Spilarctia obliqua. A dose dependent relationship of both activities was observed. The effective concentration (EC(50)) to reduce feeding and growth of the larvae of S. obliqua was found to be 617.8 and 666.9 ppm, respectively. PMID- 11933142 TI - Influence of silymarin and its flavonolignans on doxorubicin-iron induced lipid peroxidation in rat heart microsomes and mitochondria in comparison with quercetin. AB - Silymarin, an extract of Silybum marianum seeds, and the constituent flavonolignans silybin, silydianin and silychristin, as well as the flavonol quercetin, protected rat heart microsomes and mitochondria against iron-dependent doxorubicin induced lipid peroxidation. Quercetin was found to be more potent than either silymarin or its three constituents, whose cytoprotectivity was comparable. The radical scavenging activity of the compounds was investigated using a DPPH colour reduction assay and cyclic voltametry to assess their antioxidant activities. In contrast to quercetin, silybin, silydianin and silychristin did not chelate iron in aqueous solution. The results suggest that silymarin may prevent doxorubicin-mediated damage to rat heart membrane primarily through a free radical scavenging mechanism. PMID- 11933144 TI - Investigation of the larvicidal activity of Pothomorphe peltata and isolation of the active constituent. AB - The insecticidal activity of the leaves of Pothomorphe peltata (Piperaceae) was evaluated on Aedes aegypti larvae. The active methanol extract was subjected to a bioactivity-directed fractionation, monitored by the larvicidal assay, and led to the isolation of a catechol derivative, 4-nerolidylcatechol. This compound was quite potent against Aedes aegypti larvae (LC(50) = 9.1 microg/mL). PMID- 11933145 TI - Analgesic and antipyretic effects of Myrica salicifolia (Myricaceae). AB - Myrica salicifolia Hoechst (Myricaceae) root extract was found to have analgesic activity in mice. In rats there was antipyretic but no antiinflammatory activity. PMID- 11933146 TI - Antiinflammatory property of the leaves of Gongronema latifolium. AB - An aqueous extract of the dried leaves of Gongronema latifolium was studied for its antiinflammatory activity. The extract significantly (p < 0.05) inhibited carrageenan-induced rat paw oedema, carrageen-induced leucocyte migration in rats and dye leakage induced by intraperitoneal injection of acetic acid in mice. These results demonstrate the antiinflammatory property of G. latifolium. PMID- 11933147 TI - Extrahepatic biliary obstruction: can silymarin protect liver function? AB - The hepatoprotective property of silymarin is well known. However, it is not known whether the antioxidant silymarin might have a beneficial effect in extrahepatic cholestasis in common bile duct ligated rats. Malonaldehyde property concentrations, the hydrogen-donating ability and reducing power were measured in liver homogenates by spectrophotometry, as well as free SH-group levels and glutathione-reductase activities in sera. The total scavenger capacity of the livers was quantified by a chemiluminometric method. The elevated lipid peroxidation and decreased antioxidant capacity of liver homogenates and sera could be observed in ligated rats. Silymarin pretreatment improved the antioxidant capacity of the liver, diminished the direct bilirubin concentration and caused an increase of liver enzyme activities compared with the groups without treatment. These effects of silymarin suggest that it may be a useful agent for improving the antioxidant defensive system in extrahepatic cholestasis, but its choleretic property should be considered. PMID- 11933148 TI - Screening of selected basidiomycetes for inhibitory activity on Clostridium histolyticum collagenase and human leukocyte elastase. AB - Collagenase and elastase play a critical role in the degradation of connective tissue. Aqueous and dichloromethane extracts of 15 Basidiomycetes were screened for their ability to inhibit the activity of collagenase and elastase. Collagenase (EC 3.4.24.3) was not inhibited by aqueous extracts, but by dichloromethane extracts in concentrations of 200 microg/mL. For seven extracts an IC(50) less than 200 microg/mL could be observed. Elastase (3.4.21.37) was inhibited by both aqueous and dichloromethane extracts of the Basidiomycetes, with the aqueous extracts active at concentrations of 200 microg/mL. Five extracts inhibited strongly, with an IC(50) 2-20 microg/mL. Except for four, all dichloromethane extracts inhibited the enzyme in concentrations of 2 microg/mL, nine of them very strongly at about 90%. PMID- 11933149 TI - Cytotoxic activity of amooranin and its derivatives. AB - Amooranin, 25-hydroxy-3-oxoolean-12-en-28-oic acid, is a triterpene acid isolated from Amoora rohituka stem bark. The cytotoxic effects of amooranin and its derivatives were studied. Amooranin and its methyl ester showed greater cytotoxicity against MCF-7 and HeLa cells derived from tumour tissues with a 50% inhibitory concentration (IC(50)) of 1.8-3.4 microg/mL, compared with Chang liver cells from normal tissue with an IC(50) of 6.2-6.4 microg/mL, but amooranin exhibited no activity on HEp-2 and L-929 cells. However, its monoacetate derivative showed no inhibitory activity at 1-10 microg/mL dose levels. Of the cytotoxic isolates, the methyl ester derivative was inactive in in vivo evaluations in the Ehrlich ascites tumour cells at 50 and 100 mg/kg/day, demonstrating T/C values of 106% and 114%, respectively. PMID- 11933150 TI - Antiinflammatory activity of rotenoids from Clitoria fairchildiana. AB - Five structurally related rotenoids were isolated from the roots of Clitoria fairchildiana. The antiinflammatory activity was investigated using a capillary permeability assay. The protective effect was measured as the inhibition in vascular permeability produced by acetic acid in the peritoneal cavity. The results suggest that for maximum activity the maintenance of the B/C ring junction integrity is important. These findings support the traditional use of Clitoria species extracts in the treatment of inflammatory conditions. PMID- 11933152 TI - Vitamin C is one of the lipolytic substances in green tea. AB - We have studied the influence of vitamin C contained in green tea on the lipolysis of well-differentiated 3T3-L1 cells. When mature adipocytes were exposed to vitamin C the triglyceride concentration was decreased (p < 0.05) and the activity of glycerophosphate dehydrogenase, a marker of adipose conversion, was significantly inhibited (p < 0.01). These data suggest that green tea may have a lipolytic activity due to the mechanism by which the vitamin C contained in it inhibits triglyceride accumulation. PMID- 11933153 TI - Bioactivity studies on Cassia alata Linn. leaf extracts. AB - The hexane, chloroform and ethyl acetate extracts of the leaves of Cussia alata Linn. were tested for their antimutagenic, antifungal, analgesic, antiinflammatory and hypoglycaemic activities. At a dosage of 5 mg/20 g mouse, the hexane extract was analgesic as it reduced the number of squirms induced by acetic acid by 59.5%. Both the hexane and EtOAc extracts exhibited antiinflammatory activity at a dosage of 5 mg/20 g mouse with a 65.5% and 68.2% decrease in carrageenan-induced inflammation, respectively. The chloroform extract was antimutagenic, at a dosage of 2 mg/20 g mouse, with a 65.8% inhibition in the mutagenicity of tetracycline. It was also the most active against Trichophyton mentagrophytes, at a concentration of 50 mg/mL but it had no activity against Candida albicans. The hexane and EtOAc extracts showed some activity against both organisms, with the EtOAc extract being more active against C. albicans. The EtOAc extract was hypoglycaemic. At a dosage of 5 mg/20 g mouse, it decreased the blood sugar level of mice by 58.3%. Pharmacological studies showed that all extracts caused an immediate decrease in motor activity, enophthalmus, hyperemia, micturition and diarrhoea. At a dosage of 150 mg/20 g mouse, the EtOAc extract caused paralysis, screen grip loss and enophthalmus accompanied by drooping and closure of the eyelids. PMID- 11933151 TI - Causal organism of flacherie in the silkworm Antheraea assama Ww: isolation, characterization and its inhibition by garlic extract. AB - Of the different bacterial strains isolated from diseased muga silkworms, the strain named as AC-3 was found to be most pathogenic to the silkworm. Different antibiotics and plant extracts were tested for their effectiveness in inhibiting the growth of AC-3. Fresh Allium sativum (garlic extract) was most effective against the strain. The stability and MIC of the garlic extract has also been studied. We report for the first time the effectiveness of garlic extract in controlling the bacterium causing disease in the muga silkworm. PMID- 11933154 TI - The antipyretic activity of some Moroccan medicinal plants. AB - The antipyretic activity of different extracts from Calotropis procera, Cotula cinerea and Zygophyllum gaetulum was investigated experimentally in rats. The antipyretic effect was retained in all extracts tested and was comparable to that of acetylsalicylic acid used as the standard drug. PMID- 11933155 TI - Sperm axoneme: a tale of tubulin posttranslation diversity. AB - Two microtubule-containing structures are assembled during spermiogenesis: a transient manchette and a stable axoneme. Both structures contain microtubules enriched in posttranslationally modified tubulins. Despite the existence of a spectrum of tubulin isotypes postulated by the multi-tubulin hypothesis, further extended by an elaborated array of posttranslational modifications, it is unknown how this diversity influences microtubule function. There is increasing evidence that different alpha beta-tubulin isotypes can affect the structure and function of microtubules. It is also becoming increasingly clear that eukaryotic cells encode other tubulin proteins expressed by the tubulin superfamily: gamma, delta epsilon, zeta eta, and FtsZ have been identified so far. Although the role of gamma-tubulin in the nucleation of microtubule assembly is well established, the function of delta-, epsilon-, zeta-, eta-, and FtsZ-tubulins is less understood. The members of the tubulin superfamilies found in spermatids include the alpha beta-tubulin dimer, in addition to gamma-tubulin in the centrosome, and delta tubulin in the perinuclear ring region of the mouse spermatid manchette, the centrosome region, and flagellum. Posttranslational modifications in tubulin isotypes are predominant in the C-terminus exposed on the outside surface of the microtubule. This target site may influence the interaction of microtubule associated proteins, including motor proteins, and therefore determine the functional specificity of tubulin isotypes. It remains to be determined whether other newcomers to the superfamily of tubulins contain sites prone to posttranslational modification. PMID- 11933156 TI - Cdc25C expression in meiotically competent and incompetent goat oocytes. AB - Change in Cdc25C expression and localization during maturation and meiotic competence acquisition was investigated in goat oocytes. Western blot analysis revealed that Cdc25C is constitutively expressed throughout meiosis in competent goat oocytes, with changes in its phosphorylation level. Cdc25C was detected at 55 and 70 kDa, representing the nonphosphorylated form and the hyperphosphorylated active form, respectively. During the G2-M transition at meiosis resumption, Cdc25C was hyperphosphorylated as evidenced by a clear shift from 55 to 70 kDa. Okadaic acid which induced premature meiosis resumption associated with MPF activation also involved a premature shift from 55 to 70 kDa in goat competent oocytes. After artificial activation of goat oocytes, Cdc25C returned to its 55 kDa form. By indirect immunofluorescence, Cdc25C was found essentially localized in the nucleus at the germinal vesicle stage, suggesting that Cdc25C functions within the nucleus to regulate MPF activation. Concomitantly with germinal vesicle breakdown, Cdc25C was redistributed throughout the cytoplasm. The amount of Cdc25C, very low in incompetent oocytes, increased with meiosis competence acquisition. On the other hand, during oocyte growth while the expression of Cdc25C increased, its phosphorylation level increased concomitantly as well as its nuclear translocation. These results suggest that meiosis resumption needs a sufficient amount of Cdc25C which must be completely phosphorylated and nuclear and that the amount of Cdc25C may be a limiting factor for meiotic competence acquisition. We could consider that Cdc25C nuclear translocation and phosphorylation, during oocyte growth, prepare the oocytes in advance for the G2-M phase transition occurring during meiosis resumption. PMID- 11933157 TI - Identification and characterisation of known and novel transcripts expressed during the final stages of human oocyte maturation. AB - The final stages of oocyte maturation, from the germinal vesicle (GV) stage to metaphase II (MII) oocytes, are characterised by a series of dynamic events. These include germinal vesicle break down (GVBD), resumption of meiosis, and nuclear and cytoplasmic maturation to produce MII oocytes ready for fertilisation. To investigate the specific genes transcribed during these stages of oogenesis, we have prepared and analysed amplified cDNA representing the transcribed genes in a series of GV and MII oocytes. Differential display analysis disclosed that the overall gene expression profiles between different samples of GV oocytes are very similar, regardless of their source, while those between the MII oocytes are markedly variable. A comparison of expression profiles in oocytes and somatic (cumulus) cells identified several known genes preferentially-expressed in oocytes (e.g., a zona pellucida gene), as well as five novel sequences. Two of the five novel sequences are homologous to retrotransposon sequences, long terminal repeat (LTR) and long interspersed nuclear element (LINE) 1, and two other sequences show partial homology to known ESTs and genomic sequences. The remaining sequence, which is identical to shorter ESTs isolated from germ cell tumor cDNA libraries, was extended towards its 5' end by PCR, using the original cDNA preparation from which it was isolated as a template. Expression of the resultant 1.1-kb transcript is restricted to the testis and ovary, and its expression correlates with cell pluripotency in that it is expressed in embryonal carcinoma cells, but not in their differentiated derivative cells. PMID- 11933158 TI - Expression of the vascular endothelial growth factor and its receptors and effects of VEGF during in vitro maturation of bovine cumulus-oocyte complexes (COC). AB - A complete VEGF system consisting of the ligand and two of its receptors has been detected for the first time in the bovine cumulus-oocyte-complex (COC). In the course of a 24 hr in vitro maturation procedure (IVM), expression of the smaller VEGF transcripts and their specific receptors flt and flk changed remarkably in a time-dependent manner as observed by RT-PCR. The transcript concentrations of VEGF declined within 24 hr of culture, whereas both receptor mRNAs were found enriched between 6 and 12 hr of IVM. In the follicular fluid of growing ovarian follicles, immunoreactive VEGF, measured by RIA, increased significantly, reaching highest concentrations immediately before ovulation of the oocyte. The immunohistochemical localization of VEGF in bovine COCs revealed strong signals within the cumulus cell complex clearly extending beyond the oocyte cytoplasm at the beginning of in vitro maturation. After 24 hr, IVM immunoreactive VEGF disappeared remarkably from cumulus cells and the oocyte cytoplasm. An exogenous application of VEGF at the beginning of a 24 hr IVM significantly improved cleavage rates of zygotes and their development into bovine embryos showing obvious synergistic effects in combination with FSH, when compared with untreated control embryos. In addition, the number of blastomeres in deriving blastocysts increased after VEGF supplementation. These results indicate a functional VEGF system controlling important events beside the known angiogenetic effect during in vivo and in vitro maturation of the bovine COC, possibly affecting the early embryonic viability. PMID- 11933159 TI - Molecular cloning and characterization of fox testis kinectin. AB - Kinectin was isolated and characterized from a fox testis cDNA library using a monoclonal antibody (FTA-1) raised against testis surface proteins. The cDNA sequence of 4,479 nucleotides encodes an ORF of 1,330 amino acids (aa) with high homology to mouse, human, and chicken kinectins (GenBank Accession Number AF095786). Southern analysis was used to show that genes homologous to kinectin are present in several mammal species and in at least one marsupial, but not in bacteria. Alternatively spliced forms of fox kinectin were identified, and one of these is uniquely expressed in brain and spleen tissues. Kinectin expression was highest in testis relative to other tissues examined. Sequence analysis and comparisons between species revealed that kinectin encodes multiple alpha-helical coiled coils predicted to form dimers, and is, therefore, likely to exist as a dimer. The results presented in this article suggest that kinectin is required for spermatogenesis, but is not a likely candidate for use in immunocontraceptive vaccines. PMID- 11933160 TI - Expression of prepro-GnRH and GnRH receptor messengers in rainbow trout ovary depends on the stage of ovarian follicular development. AB - Gonadotropin-Releasing Hormones (GnRHs) are decapeptides well known to regulate the reproductive cycle. They are expressed not only in the brain, but also in other tissues including the gonads. It is believed that they may be involved in the endocrine and paracrine regulation of the reproductive cycle. To date, two forms of GnRH have been identified in salmonids: salmon (sGnRH) and chicken II (cGnRH-II). In the present study, the temporal expression of sGnRH-1, sGnRH-2, cGnRH-II, and rtGnRH receptor genes was studied in rainbow trout ovary during the reproductive cycle according to the stages of follicular development. Using RT PCR coupled with Southern-blot hybridization, sGnRH-1, sGnRH-2, cGnRH-II, and rtGnRH-R transcripts were detected in morphologically nondifferentiated ovaries as early as 55-65 days post-fertilization and throughout all stages of vitellogenesis. Using Northern blot analysis, cGnRH-II mRNA was detected only in immature previtellogenic ovary, whereas sGnRH mRNA was detected also during early and mid-exogenous vitellogenesis. No sGnRH mRNA was detected at the end of vitellogenesis. In maturing pre-ovulated ovary, sGnRH transiently reappeared before germinal vesicle breakdown (GVBD) and decreased thereafter. A few days after ovulation, a strong sGnRH mRNA expression was found in ovarian tissue as the eggs were kept in the body cavity of females. However, in females stripped just after ovulation, sGnRH mRNA levels remained low in ovary during several weeks. Fully spliced sGnRH-1 and sGnRH-2 messengers were mostly expressed during the reproductive cycle; however different sGnRH-1 and sGnRH-2 splicing variants containing intronic sequences were also detected. Some of these messengers may encode prepro-GnRH precursors with truncated GnRH-associated peptides. The stage dependent expression and different cell localization of sGnRH, cGnRH-II, and rtGnRH-R transcripts suggest that GnRH-like peptides may have different roles in the paracrine regulation of ovarian follicular development. PMID- 11933162 TI - Efficient method for expressing transgenes in nonhuman primate embryos using a stable episomal vector. AB - Transgenesis in the nonhuman primate can enhance the study of human biology by providing animal models for the study of primate-specific physiology, pathophysiology, and embryonic development. Progress with this technology has been hindered by the inherent inefficiency of transgenesis, transgene silencing, and practical restrictions on the production of sufficient pronuclear stage nonhuman primate zygotes. We have developed a novel technique using an Epstein Barr virus (EBV)-based episomal vector to produce rhesus monkey (Macaca mulatta) embryos expressing a transgene. Plasmid DNA containing the latent origin of replication, oriP, and Epstein Barr Nuclear Antigen-1 (EBNA-1) of EBV, as well as a CMV IE-enhanced green fluorescent protein (eGFP) expression cassette, was introduced into rhesus embryos by direct pronuclear microinjection. We detected eGFP in early cleavage stage embryos (4-8 cell) and throughout the duration of culture (day 8-9 blastocysts) by epifluorescent microscopy. A 50% transduction rate was obtained with the EBV-based vector. Microinjected embryos expressed eGFP and retained their developmental capacity as evidenced by development to the blastocyst stage. EBV-based vectors present a novel and efficient means of delivering transgenes for the study of the molecular control of primate embryonic development. PMID- 11933161 TI - A novel seminal plasma glycoprotein of a teleost, the Nile tilapia (Oreochromis niloticus), contains a partial von Willebrand factor type D domain and a zona pellucida-like domain. AB - Our previous study shows that seminal plasma of a teleost, the Nile tilapia, contains a glycoprotein Mr = 120,000 named as SPP (Seminal plasma glycoprotein)120 which forms a homopolymer that has sperm immobilizing activity. In order to elucidate the mechanisms of the formation of the homopolymer and the immobilization of sperm, molecular cloning of SPP120 was conducted. The cDNA for SPP120 contains a complete open reading frame encoding 797 amino acid residues with 14 potential N-glycosylation sites. The predicted amino acid sequence of SPP120 contains a partial von Willebrand factor type D domain and a zona pellucida domain, that are involved in protein-protein adhesion that form filamentous structures in various kinds of cells. This result suggests that SPP120 forms a homopolymer via these domains in seminal plasma and probably interacts with spermatozoa. Northern blotting reveals that the gene is also expressed in ovary, even in ovulated eggs. The results of in situ hybridization indicate that in testis the gene is expressed in Sertoli cells and epithelial cells of sperm ducts, and the localization corresponds to that of the protein analyzed by immunohistochemistry. In the ovary, the gene is expressed at the perinucleolus stage of oocytes; however, the protein is not detected in any cells other than oocytes. PMID- 11933163 TI - Nuclear and microtubule reorganization in nuclear-transferred bovine embryos. AB - We studied the nuclear and microtubule dynamics in nonactivated and pre-activated chromatin-removed oocytes following transfer of nuclei from bovine fibroblast cells. Immediately after fusion between membranes of oocytes and fibroblasts, a microtubule aster containing a gamma-tubulin spot was seen near the transferred nucleus in most oocytes regardless of activation conditions. Most fibroblast nuclei transferred into nonactivated oocytes underwent premature chromosome condensation (PCC) and divided into two masses of chromosomes. In contrast, fibroblast nuclei in pre-activated oocytes rarely underwent PCC, but formed a swelled pronuclear-like structure. Under nonactivation condition, microtubular spindles surrounded condensed chromosomes during the division of two nuclear structures. Gamma-tubulins were detected in the vicinity of condensed chromosomes, suggesting transient spindle formation. Two pronuclear-like structures near the microtubular aster containing gamma-tubulin spot(s) later formed a syngamy-like nuclear structure. While 20% of reconstructed oocytes under nonactivated conditions developed to morulae and blastocysts, only 4% of reconstructed oocytes under pre-activated conditions developed to morulae and blastocysts. These results suggest introduction of a foreign centrosome during somatic cell nuclear transfer, which probably plays a role in nuclear remodeling and subsequent development. PMID- 11933164 TI - Expression of glutamate-cysteine ligase during mouse development. AB - The tripeptide glutathione (GSH), which plays a crucial role in protecting cells against oxidative stress, is synthesized in a two-step process. The rate-limiting step is the binding of glutamate and cysteine, which is catalyzed by the enzyme glutamate-cysteine ligase (GCL). This enzyme is composed of two subunits: a large catalytic subunit (GCLc) and a smaller modifying subunit (GCLm), originating from different genes. Control of cellular GSH levels is essential for normal development. In the current study, we investigated the tissue distribution of Gclc and Gclm transcripts, as well as GCLc protein, in the developing mouse embryo. We found that both mRNAs were highly expressed in the liver and CNS at gestational day 10 (gd 10) and gd 12, with Gclm being more abundant than Gclc in the liver relative to other tissues. Also, the expression of the two subunit mRNAs was not always parallel in the embryo, in that some tissues expressed one of the subunits preferentially, suggesting that the two genes are differentially expressed during mouse development. The GCLc protein was also widely expressed throughout the embryo, and, in general, it co-localized with the Gclc mRNA. PMID- 11933165 TI - Androgen receptor and follicle-stimulating hormone receptor in the pig ovary during the follicular phase of the estrous cycle. AB - Follicle-stimulating hormone (FSH) is an important regulator of follicular development. Some effects of FSH on ovarian follicles might be enhanced by androgens. The main objectives of the present study were to examine expression of the androgen receptor (AR) and FSH receptor (FSHR) in late developing follicles in pigs. Ovaries were collected from gilts on days 13, 15, 17, and 19 of the estrous cycle (day 0 = first day of estrus, n = 4 gilts/day), a period coincident with the follicular phase. One ovary was processed for immunohistochemistry (IHC) of AR. Samples of surface wall from the largest follicles (4-5 per gilt) were dissected from the other ovary, pooled and processed for determination of AR and FSHR mRNAs using reverse transcription-polymerase chain reaction (RT-PCR). Intense AR immunostaining was present in nuclei of granulosa cells of preantral and antral follicles. AR immunoreactivity was also present in the nuclei of oocytes. Weak staining for AR was observed in cells of the theca interna, ovarian surface epithelium, and in most cells of the ovarian stroma. Relative amounts of immunoreactive AR in granulosa cells of late developing follicles, or small antral follicles (< 2 mm), did not differ between days 13, 15, 17, and 19. However, amounts of AR in granulosa cells of small antral follicles was greater (P < 0.05) than in the largest follicles present in the same ovary. The relative amounts of AR mRNA in tissue from the largest follicles on days 13, 15, 17, and 19 did not differ; however, amounts of FSHR mRNA in the same follicles were not different between days 13, 15, and 17, but decreased (P < 0.05) by day 19. Results indicate that during the follicular phase in gilts, the AR protein is mainly present in granulosa cells. Relative amounts of AR protein in granulosa cells and mRNA in walls of late developing follicles did not significantly change from day 13 to 19; however, amounts of FSHR mRNA decreased in preovulatory follicles by day 19 of the estrous cycle. PMID- 11933166 TI - Presence of angiotensin-converting enzyme in follicular fluids of porcine ovaries and its possible involvement in the intrafollicular breakdown of bradykinin. AB - The follicular fluid of porcine ovaries contains a metalloenzyme capable of hydrolyzing the synthetic substrate, benzyloxycarbonyl-Val-Lys-Met-MCA. This enzyme was purified by ammonium sulfate fractionation followed by column chromatography on DEAE-cellulose, CM-cellulose, Zn(2+)-chelating Cellulofine, and Diol-300 gel-filtration columns. The molecular weight of the purified enzyme was estimated to be 170,000 by SDS-PAGE and 400,000 by gel-filtration analysis, suggesting that the native enzyme is a dimer of the 170-kDa subunit polypeptide. The enzyme activity was drastically enhanced by the presence of chloride ion, and strongly inhibited by captopril and bradykinin potentiator B. A 9-residue peptide containing a processing site of human amyloid precursor protein was degraded by its dipeptidyl carboxypeptidase activity. Furthermore, the purified protein was recognized by specific antibody raised against human angiotensin-converting enzyme. The enzyme rapidly degraded bradykinin in vitro. These results indicate that benzyloxycarbonyl-Val-Lys-Met-MCA-hydrolyzing enzyme is a porcine angiotensin-converting enzyme, and that the enzyme may play a role in bradykinin turnover within the follicles of porcine ovaries. PMID- 11933167 TI - Inhibition of apoptosis in serum starved porcine embryonic fibroblasts. AB - In nuclear transplantation, serum starvation is a general method to synchronize donor cells at the quiescent stage (G(0)) of the cell cycle. However, serum starvation during culture of mammalian cells may induce cell death, especially through apoptosis, thus contributing to the low efficiency of nuclear transplantation. This study was performed to characterize apoptosis during serum starvation and to determine the effects of apoptosis inhibitors such as a protease inhibitor [alpha(2)-macroglobulin (MAC)] and antioxidants [N acetylcysteine (NAC), glutathione (GSH)] on serum starved porcine embryonic fibroblasts (PEF). PEF, collected from day 25-30 porcine fetuses, were cultured for 5 days in media containing 0.5% FBS to induce quiescence. Serum starved PEF showed typical morphology of apoptotic cells and stained for DNA fragmentation by TUNEL assay (26.7%). All apoptosis inhibitors tested in this study significantly (P < 0.05) reduced apoptosis of serum starved PEF, with antioxidants having better results (MAC: 7.4% vs. NAC: 1.0%, and GSH: 0.8%). Equally and importantly, the treatment with apoptosis inhibitors did not change the proportion of G(0)/G(1) stage cells. Therefore, the addition of MAC and antioxidants during serum starvation of PEF reduces apoptosis of quiescent fibroblasts and may contribute to increasing the efficiency of nuclear transplantation by improving the quality of donor nuclei. PMID- 11933168 TI - Pyruvate utilization by mouse oocytes is influenced by meiotic status and the cumulus oophorus. AB - In this study, the effects of meiotic status on the energy substrate dynamics of mouse oocyte-cumulus cell complexes (OCCs) and denuded oocytes (DOs) have been examined. In the first series of experiments, OCCs from PMSG-primed, immature mice were cultured in minimum essential medium in 8-microl microdrops under a variety of conditions, and the medium and oocytes were sampled for pyruvate and glucose concentration and for meiotic status. Oocytes in control medium underwent germinal vesicle breakdown within 3 hr and the OCCs displayed a time-dependent increase in pyruvate consumption, but the glucose concentration changed very little. Treatment with IBMX or dbcAMP, which maintained complete meiotic arrest, suppressed pyruvate consumption, but slightly more glucose was consumed than in controls. Hypoxanthine (HX) allowed up to 10% of the oocytes to resume maturation, and pyruvate and glucose consumption resembled that of control OCCs. FSH added to HX-containing medium stimulated significant glucose consumption and pyruvate production. In general, a reciprocal relationship was observed between glucose and pyruvate consumption. When the energy substrate dynamics were compared with meiotic status of the oocytes, pyruvate consumption was associated with the maturation process. Although HX maintained oocytes in the germinal vesicle stage, the meiotic arrest was "leaky," allowing increased pyruvate consumption. Additional experiments showed that DOs at either the prophase I or metaphase II stages consumed less pyruvate than oocytes actively engaged in meiotic maturation. DOs oxidized significantly more pyruvate than OCCs, and glycolytic metabolism of glucose lowered the oxidation rate in OCCs. Furthermore, while 5-6.2 times more pyruvate was consumed by OCCs than by DOs in the absence of glucose, oxidation did not mediate the meiosis-inducing effect of pyruvate, since less of this substrate was oxidized by OCCs than by DOs. We conclude that meiotically active oocytes have a greater requirement for pyruvate than prophase I- or metaphase II-arrested oocytes and that meiotic status can influence the metabolism not only of oocytes, but also of the OCCs. PMID- 11933170 TI - Association of MPF, MAPK, and nuclear progression dynamics during activation of young and aged bovine oocytes. AB - We have previously shown that bovine oocytes parthenogenetically activated after 40 hours (hr) of in vitro maturation proceed through the cell cycle faster than those after 20 hr of maturation. In the present study, we used this model of different speed of nuclear progression to investigate the correlation of two hallmarks of nuclear events, exit of metaphase arrest and pronuclear formation, with dynamics of MPF and MAPK. Bovine oocytes were matured in vitro for 20 hr (young) or 40 hr (aged) and activated in 7% ethanol followed by incubation in cycloheximide for 0, 0.5, 1, 3, 5, or 7 hr. Activity of MPF and MAPK was lower in aged than young oocytes. The responses to oocyte activation by both the two kinases and nuclear progression were faster in aged than in young oocytes. The activity of MPF declined to undetectable levels (P < 0.05) as early as 0.5 hr after activation in aged oocytes, while this did not happen in young oocytes until 3 hr after activation. The inactivation of MAPK occurred approximately 2 hr earlier in aged oocytes (5 hr post-activation) than in young oocytes (7 hr post activation). Furthermore, the decline in MPF activity preceded that of MAPK in both young and aged oocytes by about 2 hr. The decrease in activity of MPF and MAPK corresponded with the exit from meiosis and pronuclei formation regardless of the speed of nuclear progression. Despite dramatic changes in activity of MPF and MAPK, the levels of Cdc2 and Erk2 proteins were unchanged (P > 0.05) during the first 7 hr of activation. These observations suggest that inactivation of MPF and MAPK are pre-requisite for the release from metaphase arrest and formation of pronuclei in bovine oocytes. PMID- 11933169 TI - Roles of cAMP in regulation of both MAP kinase and p34(cdc2) kinase activity during meiotic progression, especially beyond the MI stage. AB - Time-dependent changes in the level of adenosine cyclic AMP (cAMP) in porcine oocytes during meiotic progression from the germinal vesicle stage (GV stage) to the metaphase II stage (MII stage) were examined using reversed-phase HPLC with UV detection. The concentration of cAMP in oocytes reached a peak at 8 hr of cultivation of cumulus-oocyte complexes (COCs), but it was dramatically decreased after 12-hr cultivation. After a 28-hr cultivation period, the level of cAMP in the oocytes had significantly reduced further, and the basal level of cAMP was observed in oocytes cultured at 32 hr and for up to 48 hr. When phosphatidylinositol 3-kinase (PI 3-kinase) or protein kinase C (PKC) in cumulus cells [which were required for meiotic progression to the MII stage in porcine oocytes (Shimada and Terada, 2001: Biol Reprod 64:1106-1114)] was suppressed by each specific inhibitor following initial 24-hr cultivation of COCs, cAMP level in the oocytes was significantly increased. After 24-hr cultivation in the maturation medium, COCs, which were cultured for an additional 24 hr in the presence of either forskolin or 3-isobutyl-1-methylxanthine (IBMX), exhibited a significant increase in the oocyte cAMP level to the similar level of that in oocytes cultured with PI 3-kinase inhibitor or PKC inhibitor, and the addition of each agent significantly suppressed meiotic progression from the MI to the MII stage and the activity of mitogen-activated protein kinase (MAPK) and p34(cdc2) kinase. These results demonstrated that when transported into oocytes from the cumulus cells via gap junctions, cAMP plays an important role not only in meiotic resumption, but also in the regulation of meiotic progression beyond the MI stage in porcine oocytes. PMID- 11933171 TI - Differential distribution of glutamylated tubulin isoforms along the sea urchin sperm axoneme. AB - Tubulin belongs to a highly conserved multigenic family, in which several gene products usually coexist in the same tissue or the same cell. Moreover, seven classes of post-translational modifications of these gene products lead to an amazing diversity of tubulin polypeptide chains, within the same cell type, whose physiological function remains elusive. Such diversity has been found in a very stable microtubular organelle, the sea urchin sperm flagellum, where some tubulin isoforms have been directly implicated in motility, whereas others may play a more structural role. In particular, polyglutamylated tubulin has been shown to be crucial for motility (Gagnon et al., 1996: J Cell Sci 109:1545 p). Here, we show with the GT335 antibody that polyglutamylated tubulin is distributed according to a decreasing gradient along the sea urchin sperm axoneme, since a semi-quantitative measurement of immunofluorescence intensity reveals that in its proximal half, the axoneme is sixfold more labeled than in its distal half. This gradient along the length of the axoneme is confirmed by immunogold labeling procedures which, in addition, demonstrate a uniform distribution of polyglutamylated tubulin among peripheral doublets and a lesser content in the central pair within a same section. Moreover, our data obtained with B3, an antibody that recognizes both mono- and poly-glutamylated tubulin, suggest that the number of glutamate residues in the lateral poly-glutamyl chain of tubulin varies along the whole length of the axoneme. These novel results coupled with those published earlier may be important to understand the role of polyglutamylation in flagellar motility. PMID- 11933172 TI - Squamous cell carcinoma of the head and neck in solid organ transplant recipients. AB - BACKGROUND: The increased incidence of cancer after solid organ transplantation is well established in the literature, yet outcome studies in this population are rare. Excluding skin cancers, squamous cell carcinomas make up most head and neck cancers in transplant recipients. METHODS: At our institution, of 5300 solid organ transplant recipients, 34 have had head and neck cancer develop. We reviewed the records of the 23 recipients whose cancer was treated here. RESULTS: Only 6 of the 23 recipients were alive at the time of our chart review. Of these, three had already survived 5 years. The 10 recipients diagnosed early (stage I or II) had significantly longer survival after cancer diagnosis than the 13 diagnosed at an advanced stage (stage III or IV) (96.0 mo vs 9.0 mo, p <.001). In all, 14 (60.8%) of the 23 recipients died of cancer within 2 years after diagnosis, 12 (50.2%) within 12 months. The sum of the daily doses of immunosuppressive drugs at cancer diagnosis was significantly greater for recipients who died within 2 years (p =.02). Furthermore, the difference in average doses of both prednisone (p =.001) and azathioprine (p =.028) was also significantly greater for those who died within 2 years. The average dose of cyclosporine was also greater, but this difference did not reach statistical significance (p =.18). The average dose of prednisone was significantly lower for recipients diagnosed early (p =.001). This correlation between high immunosuppressive drug doses and worse outcome has not been shown previously. CONCLUSIONS: Solid organ transplant recipients who are diagnosed with advanced head and neck cancer while receiving high doses of immunosuppressive drugs fare extremely poorly. High doses of immunosuppressive drugs, most notably prednisone, correlate significantly with advanced diagnosis of head and neck cancer and earlier death. PMID- 11933173 TI - Omental free flap reconstruction in complex head and neck deformities. AB - BACKGROUND: Microvascular free flaps continue to revolutionize coverage options in head and neck reconstruction. This article reviews our 25-year experience with omental free tissue transfers. METHODS: All patients who underwent free omental transfer to the head and neck region were reviewed. RESULTS: Fifty-five patients were included with omental transfers to the scalp (25%), craniofacial (62%), and neck (13%) region. Indications were tumor resections, burn wound, hemifacial atrophy, trauma, and moyamoya disease. Average follow-up was 3.1 years (range, 2 months-13 years). Donor site morbidities included abdominal wound infection, gastric outlet obstruction, and postoperative bleeding. Recipient site morbidities included partial flap loss in four patients (7%) total flap loss in two patients (3.6%), and three hematomas. CONCLUSIONS: The omental free flap has acceptable abdominal morbidity and provides sufficient soft tissue coverage with a 96.4% survival. The thickness ?and versatility of omentum provide sufficient contour molding for craniofacial reconstruction. It is an attractive alternative for reconstruction of large scalp defects and badly irradiated tissue. PMID- 11933174 TI - Glomus jugulare tumor: tumor control and complications after stereotactic radiosurgery. AB - BACKGROUND: We evaluated toxicity and long-term efficacy of stereotactic radiosurgery in patients with symptomatic or progressive glomus jugulare tumors. METHODS: Twenty-five consecutive patients (age, 30-88 years; 17 women, 8 men) who underwent stereotactic radiosurgery with the Leksell Gamma Knife (dose, 12-18 Gy) were prospectively followed. MRI and clinical examinations were performed at 6 months and 1, 2, and 3 years, and then every 2 years. RESULTS: None of the tumors increased in size, 17 were stable, and 8 decreased (median imaging follow-up, 35 months; range, 10-113 months). Symptoms subsided in 15 patients (60%); vertigo occurred in 1, but balance improved with vestibular training (median clinical follow-up, 37 months; range, 11-118 months). No other new or progressive neuropathy of cranial nerves V-XII developed. CONCLUSIONS: Stereotactic radiosurgery can achieve excellent tumor control with low risk of morbidity in the treatment of glomus jugulare tumors. The lower cranial nerves can safely tolerate a radiosurgical dose of 12 to 18 Gy. PMID- 11933175 TI - Results of the treatment of locally invasive thyroid carcinoma. AB - BACKGROUND: Well-differentiated thyroid carcinoma usually has an excellent prognosis. However, when extrathyroidal invasion occurs, it is associated with significant morbidity and mortality. This report presents the experience of a single institution in the treatment of patients with locally invasive, well differentiated thyroid carcinoma. PATIENTS AND METHODS: Forty-six patients with locally invasive well-differentiated thyroid carcinoma were diagnosed. Histopathologic types included: 28 papillary carcinoma and 18 follicular. RESULTS: Patients with exclusive invasion of the muscle or recurrent laryngeal nerve usually had complete tumor resection. Patients with tracheal, laryngeal, or esophageal invasion usually underwent shave resection. The factors that adversely affected survival were: age >45 years, preoperative diagnosis of extrathyroidal extension, and incomplete resection (p <.05). CONCLUSIONS: There were similar survival results after complete or shave resection and poor survival when the resection was incomplete. Tumors with minimal invasion can be treated by shave resection with acceptable survival and low morbidity. PMID- 11933176 TI - Is there a role for positron emission tomography with 18F-fluorodeoxyglucose in the initial staging of nodal negative oral and oropharyngeal squamous cell carcinoma. AB - BACKGROUND: The aim of our study was to assess the value of positron emission tomography (PET) with 18F-fluorodeoxyglucose (FDG) for the staging of clinically nodal negative necks in oral and oropharyngeal squamous cell carcinoma (SCC) using sentinel lymph node (SLN) biopsy and elective neck dissection (END) as "gold standard" for comparison. METHODS: Twelve patients (10 men, 2 women) with oral or oropharyngeal squamous cell carcinoma and no evidence of lymph node metastasis in the physical and radiologic examinations were eligible. At least 24 hours before surgery PET with FDG were performed. The SLN was localized by preoperative lymphoscintigraphy with 99m-Tc-Nanocoll and intraoperative use of a hand-held gamma-probe and selectively excised. All patients then underwent END. RESULTS: SLN and END revealed occult metastasis in 5 of 12 cases. SLN biopsy was accurately feasible in all 12 patients and diagnosed all 5 cases of occult metastasis (sensitivity and specificity of 100%). PET suggested two patients having occult metastasis, of which one turned out to be false positive (sensitivity 25%, specificity 88%). The mean size of the micrometastasis was 1.4 mm (range, 1.2-1.5 mm). CONCLUSIONS: PET with FDG turned out to have a poor sensitivity and specificity in revealing occult metastasis and has no role for the evaluation of otherwise clinically N0 necks. The failure to detect micrometastasis by PET is due to the technical limitations of resolution (4-5 mm). SLN biopsy, with END in cases of positive SLN, provides a highly accurate staging of N0 necks in oral and oropharyngeal carcinoma. Patients with negative SLN could be spared the risks and the morbidity of END. PMID- 11933177 TI - Control of regional metastasis after induction chemotherapy and radiotherapy for nasopharyngeal carcinoma. AB - BACKGROUND: To study the impact of adding induction chemotherapy to radiotherapy on the long-term control of regional metastasis and survival in patients with nasopharyngeal carcinoma (NPC). METHODS: Between February 1988 and August 1993, 240 NPC patients with Ho's T3 stage, N2-3 stage, or nodal size >/=3 cm were recruited onto two randomized trials comparing induction chemotherapy followed by radiotherapy (CT + RT) and radiotherapy alone (RT) using a similar treatment protocol. Of these, 210 patients (105 in each treatment arm) had cervical nodal metastasis and were included in the analysis. Patients in the CT + RT arm received two to three cycles of cisplatin, 60 mg/m(2) day 1, + epirubicin, 110 mg/m(2) day 1, followed by radiotherapy. Radiotherapy technique and dose were similar in both arms. The median follow-up time was 71 months (range, 5-152 months). RESULTS: The overall response rate of nodal disease to chemotherapy was 86%, and the complete response (CR) rate was 44%. At the end of radiotherapy, 92% of patients in the CT + RT arm and 86% in the RT arm achieved CR in the neck (p =.12). The 5-year nodal relapse-free survival rates in the CT + RT and RT arm were 83% and 75%, respectively (p =.13). Most neck failures (81%) occurred during the first 36 months of follow-up. Radical neck dissection successfully salvaged 41% of neck failures in the CT + RT arm and 46% in the RT arm. The 5-year distant metastases-free survival rates were 70% in the CT + RT arm and 68% in the RT arm (p =.56), and the corresponding 5-year disease-specific survival rates were 66% and 68%, respectively (p =.55). In subgroup analysis, no significant differences in regional control and survival could be found in patients with Ho's N2-3 stage, AJCC N2-3 stage, or nodal size >6 cm. CONCLUSIONS: Induction chemotherapy does not seem to improve the regional control and survival in NPC patients with regional metastasis compared with radiotherapy alone and is not recommended as a routine treatment outside the context of a clinical trial. PMID- 11933178 TI - Cervical nodal metastases from occult primary: undifferentiated carcinoma versus squamous cell carcinoma. AB - PURPOSE/OBJECTIVE: Controversy exists regarding the management of cervical lymph node metastases from occult primary. Oncologists face a major challenge in adopting an optimal approach. This study attempted to compare the clinical course of two different histologic findings of this disease entity. MATERIALS AND METHODS: A retrospective analysis was performed for all patients referred to our institution between 1988 and 1998 with cervical lymph node metastases from an unknown primary. Case records of consecutive unselected patients with histologically confirmed carcinoma in cervical lymph nodes were reviewed. Those with histologic findings other than squamous cell carcinoma (SCC) or undifferentiated carcinoma (UDC) and lymphadenopathies at the supraclavicular fossa alone or below the clavicles at the time of diagnosis were excluded. There were 45 patients identified with a mean follow-up of 36 months (range, 4-110 months). Thirty-seven were men and eight were women. The mean age was 57 (range, 29-91). There were 32 patients with SCC and 13 patients with UDC. Treatment modality included surgery (S) alone in 1 patient (2%), radiotherapy (RT) alone in 24 patients (53%), and combined modality in 20 patients (45%). (Twelve patients (27%) had combined S and RT, 8 patients (18%) had combined chemotherapy and RT.) Twenty-eight patients (62%) were treated with radical intent. For those patients treated by radical RT, the RT field covered both sides of the neck and the potential mucosal primary (PMP) sites, including the entire pharyngeal axis. The median radiation doses to the lymph nodes and the PMP were 65 Gy (range, 60-70 Gy) and 60 Gy (range, 40-70 Gy), respectively. RESULTS: At the time of analysis, ultimate control of disease above the clavicles according to N stage, treatment intent, and histologic type was as follows: N1s, 7 of 7 (100%); N2s, 15 of 26 (58%); N3s, 1 of 12 (8%); radical intent, 19 of 28 (68%); palliative intent, 3 of 17 (18%); UDC, 11 of 13 (85%); SCC,11 of 32 (34%). Eleven patients remained alive and disease free, with a median follow-up of 79 months (range, 27-110 months). The 5-year disease-specific survival (DSS) for the radical treatment group and the palliative treatment group were 67% and 18%, respectively (p =.0011). Significant difference in 5-year DSS was observed among the different N groups: 100% for N1s, 55% for N2s, and 0% for N3s, respectively (p =.0001). There was also a significant difference in the 5-year DSS between UDC and SCC: 81% for UDC vs 34% for SCC (p =.01). No significant difference in the 5-year DSS was observed on the basis of treatment modality in the radically treated group: 63% for RT alone vs 75% for S + RT (p =.711). CONCLUSIONS: UDC histologic findings in our series are associated with better locoregional control and DSS than SCC. Our results in local control, emergence of primary tumor, and DSS are comparable with other published data. However, disease control of advanced nodal stage remains poor; more aggressive treatment approaches, like the use of concurrent chemoradiation or altered fractionation scheme, should be explored. PMID- 11933179 TI - Patterns of invasion and routes of tumor entry into the mandible by oral squamous cell carcinoma. AB - BACKGROUND: An understanding of the patterns, spread, and routes of tumor invasion of the mandible is essential in deciding the appropriate level and extent of mandibular resection in oral squamous cell carcinoma. METHODS: A prospective study of histologic patterns of tumor invasion and routes of tumor entry into the mandible was performed in a consecutive series of 100 previously untreated patients. RESULTS: The pattern of tumor invasion of the mandible depended on the depth of invasion both in the hard (p =.001) and soft tissues (p =.001). There was evidence that the pattern of invasion was related to histologic prognostic indicators of the disease, such as extracapsular spread from invaded lymph nodes (p =.03). The route of tumor entry was at the point of abutment to the mandible (direct) in all 13 cases, invading the dentate part of the mandible. Fifty-five percent (23 of 42) of tumors invading the edentulous ridge entered through the occlusal (superior) surface. Direct entry to the mandible in the edentulous ridge was more likely for tumors arising in the tongue, floor of the mouth and the buccal mucosa compared with alveolar or retromolar sites (p =.003) CONCLUSIONS: Larger or more deeply invading tumors in the soft tissue are more likely to invade the mandible and show the more aggressive (invasive) form of tumor spread, reducing the options of a more conservative (rim) resection. Tumors tend to enter the mandible at the point of abutment, which in both the dentate and edentulous jaw is often at the junction of the reflected and attached mucosa. A point of tumor entry below the occlusal ridge or gingival crest should be assumed when planning rim or marginal resections of the mandible. PMID- 11933180 TI - Radiosurgery for paraganglioma of the temporal bone. AB - PURPOSE: To report the outcome of patients with paraganglioma of the temporal bone treated with stereotactic radiosurgery at the University of Florida. METHODS AND MATERIALS: Between January 1997 and June 1999, five patients with paraganglioma of the temporal bone were treated with Linac-based stereotactic radiosurgery at the University of Florida. The ages of the three female and two male patients were between 40 and 88 years (median, 49 years). Four patients were treated at initial presentation, and one had recurrent disease. Treatment volumes ranged from 4.9 cm3 to 18.4 cm3, with a mean of 10.84 cm3. The dose applied to the margin of the tumor varied from 12.5 to 15 Gy (median, 15 Gy). The treatment dose was specified to the 80% isodose shell in two cases and to the 70% isodose shell in three cases. The median follow-up time was 27 months, ranging from 14 to 50 months. RESULTS: One of four previously untreated patients had a relapse at the primary tumor site. Treatment failure occurred at the field margin 6 months after radiosurgery; the patient was subsequently treated with fractionated stereotactic radiotherapy and at the time of analysis had no evidence of disease, 21 months after initiation of salvage therapy. The patient treated at the time of recurrence after conventional radiotherapy had a local recurrence 40 months after radiosurgery. At the time of this recurrence, the patient had biopsy-proven metastatic disease in two cervical lymph nodes, and no salvage therapy was performed. All patients were alive at the time of the analysis, one with disease present. Presenting symptoms improved in two patients and stabilized in one. The two patients who had local recurrence develop had worsening of their symptoms. One patient had a cranial nerve V palsy develop 6 months after treatment, which resolved after a few months. CONCLUSIONS: In this series, the results with stereotactic radiosurgery are discouraging compared with our results with conventional fractionated radiotherapy in patients with paraganglioma of the temporal bone. PMID- 11933181 TI - Radiotherapy for carcinoma in situ of the true vocal cords. AB - PURPOSE: To report long-term rates of tumor control after radiotherapy (RT) for carcinoma in situ (CIS) of the true vocal cords (TVC). A comprehensive literature review was performed, and outcomes with other modalities of treatment for CIS of the TVC were compared. METHODS AND MATERIALS: Thirty patients with CIS of the TVC were treated between July 1967 and May 1998 with curative intent using megavoltage RT. Most patients (28 of 30) were treated with cobalt-60 through small (usually 5 x 5 cm) fields. Median RT dose was 56.25 Gy (range, 56.25-75 Gy; mean dose, 59.15 Gy) at 2.25 Gy per fraction. One patient was treated for a synchronous head and neck primary malignancy with large fields to a total dose of 75 Gy at 1.8 Gy per fraction. Approximately two thirds of the patients (19 of 30) were referred for RT because of recurrence after at least one stripping procedure. RESULTS: With a mean follow-up of 7.1 years (range, 2-17 years), the 5 year rates of local control, local control with larynx preservation, and ultimate local control (including salvage surgery) were the following: 88%, 88%, and 100%, respectively. Invasive squamous cell carcinoma developed in three (10%) of the patients. Time to failure was 14 months, 34 months, and 48 months, respectively. All three patients were surgically salvaged with a total laryngectomy. Cause specific survival at 5 years was 100%. There were no late complications. CONCLUSIONS: RT to approximately 60 Gy at 2.25 Gy per fraction using small (5 x 5 cm) fields produces excellent results with CIS of the TVC. PMID- 11933182 TI - The Wookey flap revisited. AB - BACKGROUND: Complete circumferential pharyngoesophageal reconstruction has undergone remarkable changes and developments during the last three decades. Gastric transposition and microvascular techniques are the standards of care. However, in cases of failure, or where other factors such as severe comorbidities prevent the use of microvascular techniques, reconstruction with local flaps provide a reliable option. We have used the Wookey flap technique in six patients with complete circumferential loss of the pharynx and upper esophagus. To make the procedure more reliable, we added a "delay" stage. METHODS: Retrospective review of charts of six Wookey-type reconstructions of the pharynx and upper esophagus done in our department at SUNY-HSC, Brooklyn, and The Long Island College Hospital. RESULTS: Five of the six patients had successful reconstruction of complete circumferential pharyngoesophageal resection. All five resumed oral alimentation. The flap failed in one patient. CONCLUSIONS: The Wookey flap may still provide surgeons with a "last resort" reconstructive option after complete, circumferential pharyngoesophageal resection. PMID- 11933183 TI - Metastatic cancer to the floor of mouth: the lingual lymph nodes. AB - BACKGROUND: The upper level of a cervical lymphadenectomy is anatomically defined at its anterior extent by the lower border of the mandible and, in surgical practice, by the lingual nerve. A neck dissection completed below this level is generally considered adequate for removal of lymph nodes at risk for metastases from oral cavity cancer. Traditional discontinuous neck dissections do not provide for removal of floor of mouth tissue along with the primary and neck specimens. METHODS: A case report presenting biopsies from a T2N2bM0 squamous cell carcinoma of the mobile tongue and adjacent floor of the mouth in a 73-year old man. RESULTS: Deep biopsy of a ventral tongue and floor of mouth squamous cell carcinoma revealed occult metastatic cancer to lymph nodes located in the superficial floor of mouth associated with the sublingual gland above the lingual nerve. This report identifies floor of mouth lymph nodes that can be involved with cancer and missed through the standard practice of discontinuous neck dissection.Conclusions. This finding offers evidence that, in certain cases, a traditional discontinuous neck dissection may not address all lymph nodes at risk in the treatment of oral cavity cancer. Further investigation into lymph node distribution within the oral cavity is warranted to reappraise the upper limits of cervical lymphadenectomy. PMID- 11933184 TI - Response to paclitaxel and carboplatin in metastatic salivary gland cancer: a case report. AB - BACKGROUND: Malignant tumors of the salivary gland are rare entities that are treated primarily by surgical resection. For patients with recurrent or unresectable disease, options include radiation therapy or chemotherapy; however, responses are few and of short duration. Patients with metastatic disease have been treated with chemotherapy, but, again, response rates have been low and of short duration. METHODS: A 52-year-old man was seen with a mass on his tongue. A biopsy revealed adenocarcinoma of a minor salivary gland. Ten months after surgical resection, neck dissection, and radiation therapy, the patient was found to have metastatic disease to the lung. Chemotherapy was initiated with carboplatin and paclitaxel. RESULTS: The patient obtained a complete response after six cycles of carboplatin and paclitaxel. CONCLUSIONS: The use of carboplatin and paclitaxel in the setting of metastatic salivary gland cancer is a viable option. PMID- 11933185 TI - Mutation detection 2001: Novel technologies, developments and applications for analysis of the human genome. Proceedings of the 6th International Symposium on Mutations in the Human Genome. Bled, Slovenia, 2001. PMID- 11933186 TI - SNP genotyping with fluorescence polarization detection. AB - When a fluorescent molecule is excited by plane polarized light, the fluorescence emitted is also polarized. The degree of fluorescence polarization (FP) detected, under constant temperature and solvent viscosity, is proportional to the molecular weight of the dye molecule. By monitoring the FP of a fluorescent dye, one can detect significant changes in the molecular weight of the molecule without separation or purification. Because the size of the probe is altered in the course of a number of single nucleotide polymorphism (SNP) genotyping reactions, FP is therefore an excellent detection mechanism for these assays. Indeed, FP detection can be used in SNP genotyping with the primer extension TaqMan((R)) and Invader((R)) assays. Use of FP detection makes it possible to reduce the cost of TaqMan((R)) and Invader((R)) probes by abrogating the need for a fluorescence quencher. Moreover, inexpensive, unpurified, and unlabeled probes are used in the primer extension reaction with FP detection. As an end-point detection mechanism, FP detection is suitable for high-throughput SNP genotyping. PMID- 11933187 TI - Commercial molecular diagnostics in the U.S.: The Human Genome Project to the clinical laboratory. AB - Molecular diagnosis is the detection of pathogenic mutations in DNA and RNA samples to aid in detection, diagnosis, subclassification, prognosis, and monitoring response to therapy. Principles underlying nucleic-based diagnosis originate from localization, identification, and characterization of genes responsible for human disease. Clinical molecular genetics is now part of the mainstream of medical care in the United States. All commercial clinical reference laboratories now have a molecular genetic diagnostic unit, many of which are in contractual agreement with third party payers to provide services. Gene discovery provides valuable insight into the mechanisms of disease processes and gene-based markers will enable clinicians to study disease predisposition, as well as improved methods for diagnoses, prognosis, and monitoring of therapy. The broad range of mutation spectrum and type performed in the clinical laboratory requires the use of multiple technologies rather than a single typing platform. Platform choice depends on such diverse factors as local expertise, test volume, economies of scale, R&D budget, and royalties. Test validation is a major hurdle and positive control samples are often not readily available. Oversight and the regulatory environment for clinical molecular genetics laboratories in the United States are evolving rapidly. Several government agencies and private organizations are currently involved in revision of specific laboratory standards, including the Secretary's Advisory Committee on Genetic Testing (SACGT), Food and Drug Administration (FDA), Center for Disease Control (CDC), College of American Pathologists (CAP), American College of Medical Genetics (ACMG), and the individual states. PMID- 11933188 TI - An update on conformation sensitive gel electrophoresis. AB - Conformation-sensitive gel electrophoresis (CSGE) was developed as a method of heteroduplex analysis to screen large multi-exon genes for sequence variation. The novelty of the method was in the use of a non-proprietary acrylamide gel matrix that used 1,4-bis (acrolyl) piperazine (BAP) as a cross linker with ethylene glycol and formamide as mildly denaturing solvents. The denaturing environment enhances the conformation polymorphism present in DNA heteroduplexes containing variations as small as single nucleotide polymorphisms (SNPs). CSGE has also been adapted for use on a fluorescent platform (F-CSGE) that resulted in higher throughput and sensitivity. Variation in sensitivity of CSGE has been studied extensively. The results demonstrate that the nature of the mismatched base in a defined sequence context has the most profound effect on the conformation of the heteroduplex. Additionally, the size of the PCR product, as well as the location of the mismatch within the PCR product, are two important parameters that determine the resolution of the mismatch-containing heteroduplexes during CSGE. Like any other mutation scanning technique, CSGE can have limited resolution of two closely linked sequence variations. For specific genes, like BRCA1 and BRCA2 where multiple SNPs are present in the coding sequence, each CSGE shift has to be sequenced to define the exact nature of the sequence change. In conclusion, CSGE scanning provides a powerful, cost-efficient way to scan genes with high sensitivity and specificity. PMID- 11933189 TI - Analysis of SNPs and other genomic variations using gel-based chips. AB - Application of microarrays for the analysis of point mutations and SNPs in genomic DNAs is currently under intensive development. Various technologies are being investigated, employing enzymatic, chemical, and physical tools [for review, see Tillib and Mirzabekov, 2001]. Our current approach is based on the use of IMAGE chips (immobilized microarrays of gel elements) consisting of an array of gel pads attached to a hydrophobic glass surface. The gel pads range in size from picoliters to nanoliters and are used for immobilization of oligonucleotide probes, as well as miniature test tubes for chemical or enzymatic reactions with tethered compounds. Nucleic acids are hybridized, fractionated, modified, and subjected to enzymatic reactions inside the pads. All steps of sequence analysis (PCR-amplification, activation or release of primers and products, DNA extension, hybridization, and reading of the results) can be performed within the same pad. A flexible and inexpensive technology platform enables one to monitor processes in the arrays in both real time and steady state. Identification of SNPs, microsequencing, and other specific tasks are easily performed. In particular, stacking interactions with short oligonucleotides enhance the capability of high-throughput screening. The IMAGE chips can be analyzed using a variety of equipment, from a dedicated multi-color fluorescent microscope or MALDI-spectrometer to an inexpensive portable analyzer suitable for field conditions. Customized gel-based chips were successfully used for screening of SNPs in a broad range of biologically meaningful genes. PMID- 11933190 TI - Determining sequence length or content in zero, one, and two dimensions. AB - High-throughput assays are essential for the practical application of mutation detection in medicine and research. Moreover, such assays should produce informative data of high quality that have a low-error rate and a low cost. Unfortunately, this is not currently the case. Instead, we typically witness legions of people reviewing imperfect data at astronomical expense yielding uncertain results. To address this problem, for the past decade we have been developing methods that exploit the inherent quantitative nature of DNA experiments. By generating high-quality data, careful DNA-signal quantification permits robust analysis for determining true alleles and certainty measures. We will explore several assays and methods. In a one-dimensional readout, short tandem repeat (STR) data display interesting artifacts. Even with high-quality data, PCR artifacts such as stutter and relative amplification can confound correct or automated scoring. However, by appropriate mathematical analysis, these artifacts can be essentially removed from the data. The result is fully automated data scoring, quality assessment, and new types of DNA analysis. These approaches enable the accurate analysis of pooled DNA samples, for both genetic and forensic applications. On a two-dimensional surface (comprised of zero dimensional spots) one can perform assays of extremely high-throughput at low cost. The question is how to determine DNA sequence length or content from nonelectrophoretic intensity data. Here again, mathematical analysis of highly quantitative data provides a solution. We will discuss new lab assays that can produce data containing such information; mathematical transformation then determines DNA length or content. PMID- 11933191 TI - DHPLC screening of cystic fibrosis gene mutations. AB - Denaturing high performance liquid chromatography (DHPLC) using ion-pairing reverse phase chromatography (IPRPC) columns is a technique for the screening of gene mutations. In order to evaluate the potential utility of this assay method in a clinical laboratory setting, we subjected the PCR products of 73 CF patients known to bear CFTR mutations to this analytic technique. We used thermal denaturation profile parameters specified by the MELT program tool, made available by Stanford University. Using this strategy, we determined an initial analytic sensitivity of 90.4% for any of 73 known CFTR mutations. Most of the mutations not detected by DHPLC under these conditions are alpha-substitutions. This information may eventually help to improve the MELT algorithm. Increasing column denaturation temperatures for one or two degrees above those recommended by the MELT program allowed 100% detection of CFTR mutations tested. By comparing DHPLC methodology used in this study with the recently reported study based on Wavemaker 3.4.4 software (Transgenomic, Omaha, NE) [Le Marechal et al., 2001) and with previous SSCP analysis of CFTR mutations [Ravnik-Glavac et al., 1994] we emphasized differences and similarities in order to refine the DHPLC system and discuss the relationship to the alternative approaches. We conclude that the DHPLC method, under optimized conditions, is highly accurate, rapid, and efficient in detecting mutations in the CFTR gene and may find high utility in screening individuals for CFTR mutations. Hum Mutat 19:374-383, 2002. Published 2002 Wiley-Liss, Inc. PMID- 11933192 TI - Correlation of MFOLD-predicted DNA secondary structures with separation patterns obtained by capillary electrophoresis single-strand conformation polymorphism (CE SSCP) analysis. AB - We have studied 57 different mutations within three beta-globin gene promoter fragments with sizes 52 bp, 77 bp, and 193 bp by fluorescent capillary electrophoresis CE-SSCP analysis. For each mutation and wild type, energetically most-favorable predicted secondary structures were calculated for sense and antisense strands using the MFOLD DNA-folding algorithm in order to investigate if any correlation exists between predicted DNA structures and actual CE migration time shifts. The overall CE-SSCP detection rate was 100% for all mutations in three studied DNA fragments. For shorter 52 bp and 77 bp DNA fragments we obtained a positive correlation between the migration time shifts and difference in free energy values of predicted secondary structures at all temperatures. For longer 193 bp beta-globin gene fragments with 46 mutations MFOLD predicted different secondary structures for 89% of mutated strands at 25 degrees C and 40 degrees C. However, the magnitude of the mobility shifts did not necessarily correlate with their secondary structures and free energy values except for the sense strand at 40 degrees C where this correlation was statistically significant (r = 0.312, p = 0.033). Results of this study provided more direct insight into the mechanism of CE-SSCP and showed that MFOLD prediction could be helpful in making decisions about the running temperatures and in prediction of CE-SSCP data patterns, especially for shorter (50-100 bp) DNA fragments. PMID- 11933193 TI - Large-scale genotyping of single nucleotide polymorphisms by Pyrosequencingtrade mark and validation against the 5'nuclease (Taqman((R))) assay. AB - Here we present the first large-scale effort at genotyping using a novel sequencing method, Pyrosequencingtrade mark, as a method for genotyping of single nucleotide polymorphisms (SNPs). Pyrosequencingtrade mark genotypes were validated through duplicate analysis of 1,022 genotypes using the PSQ96trade mark instrument for pyrosequencing and TaqMan((R)) for 5'nuclease assays. Identical results were obtained using both methods. In a small pilot study, a pooling strategy using Pyrosequencingtrade mark was successfully tested. We conclude that Pyrosequencingtrade mark is highly efficient and accurate in the analysis of SNPs and represents a promising solution to high-throughput genotyping of large sample populations. PMID- 11933194 TI - New developments in high-throughput resequencing and variation detection using high density microarrays. AB - We developed a high-throughput method for resequencing for single nucleotide polymorphism (SNP) discovery using high-density microarrays. Over the two-year course of this study a number of improvements in sample preparation methods, hybridization assay, array handling, and analysis method were developed and implemented. DNA from 40 unrelated individuals of three different ethnic origins was amplified, labeled, and hybridized to arrays designed with probes representing genomic, coding, and regulatory regions. Protocol improvements including the use of long PCR and semi-automation reduced labeling and fragmentation costs by 33%. Automation improvements include the development of a scanner autoloader for arrays, a faster array wash station, and a linked laboratory tracking and data management system. Validation of a smaller feature size, 20 x 24 microns, allowed the simultaneous screening of 30-kb sense and 30 kb antisense DNA on each microarray, increasing throughput to 1.4 Mb per day per two laboratory personnel. More than 15,000 SNPs were identified in 8.3 Mb of the human genome using high-density resequencing and variation detection arrays (microarrays). PMID- 11933195 TI - Making ends meet in genetic analysis using padlock probes. AB - Padlock probes are molecular tools that combine highly specific target sequence recognition with the potential for multiplexed analysis of large sets of target DNA or RNA sequences. In this brief review, we exemplify the ability of these probes to distinguish single-nucleotide target sequence variants. We further discuss means to detect the location of target sequences in situ, and to amplify reacted padlock probes via rolling-circle replication, as well as to sort reaction products on tag-arrays. We argue that the probes have the potential to render high-throughput genetic analyses precise and affordable. PMID- 11933196 TI - A surface invasive cleavage assay for highly parallel SNP analysis. AB - The structure-specific invasive cleavage of single-stranded DNA by 5' nucleases is a useful means for sensitive detection of single-nucleotide polymorphisms or SNPs. The solution-phase invasive cleavage reaction has sufficient sensitivity for direct detection of as few as 600 target molecules with no prior target amplification. One approach to the parallelization of SNP analysis is to adapt the invasive cleavage reaction to an addressed array format. Two surface invasive cleavage reaction strategies were designed and tested using the polymorphic site in codon 158 of the human ApoE gene as a model system, with a synthetic oligonucleotide as target. The upstream oligonucleotide, which is required for the invasive cleavage reaction, was either added in solution (strategy 1), or co immobilized on the surface along with the probe oligonucleotide (strategy 2). Both strategies showed target-concentration and time-dependent amplification of signal. Parameters that govern the rate of the surface-invasive cleavage reactions are discussed. PMID- 11933197 TI - Myotonia caused by mutations in the muscle chloride channel gene CLCN1. AB - Pure non-syndromic, non-dystrophic myotonia in humans is caused by mutations in the genes coding for the skeletal muscle sodium channel (SCN5A) or the skeletal muscle chloride channel (CLCN1) with similar phenotypes. Chloride-channel myotonia can be dominant (Thomsen-type myotonia) or recessive (Becker-type myotonia). More than 60 myotonia-causing mutations in the CLCN1 gene have been identified, with only a few of them being dominant. A common phenotype of dominant mutations is a dominant negative effect of mutant subunits in mutant-WT heterodimers, causing a large shift of the steady-state open probability voltage dependence towards more positive, unphysiological voltages. The study of the properties of disease causing mutations has helped in understanding the functional properties of the CLC-1 channel that is part of a nine-member gene family of chloride channels. The large body of knowledge obtained for CLC-1 may also help to better understand the other CLC channels, three of which are also involved in genetic diseases. PMID- 11933198 TI - Germline BRCA1 promoter deletions in UK and Australian familial breast cancer patients: Identification of a novel deletion consistent with BRCA1:psiBRCA1 recombination. AB - Inherited susceptibility to breast cancer results from germline mutations in one of a number of genes including BRCA1. A significant number of BRCA1-linked familial breast cancer patients, however, have no detectable BRCA1 mutation. This could be due in part to the inability of commonly used mutation-detection techniques to identify mutations outside the BRCA1 coding region. This paper addresses the hypothesis that non-coding region mutations, specifically in the BRCA1 promoter, account for some of these cases. We describe a new and detailed restriction map of the 5' region of the BRCA1 gene including the nearby NBR2, psiBRCA1, and NBR1 genes and the isolation of a number of new informative hybridization probes suitable for Southern analysis. Using this information we screened DNA from lymphoblastoid cell-lines made from 114 UK familial breast cancer patients and detected one large deletion in the 5' region of BRCA1. We show that the breakpoints for this deletion are in BRCA1 intron 2 and between NBR2 and exon 2 of psiBRCA1, raising the possibility that this deletion arose via a novel mechanism involving BRCA1:psiBRCA1 recombination. We have also screened 60 familial breast cancer patients from the Australian population, using an amplification refractory mutation system (ARMS) technique described previously by our group, and found one patient with a genotype consistent with a BRCA1 promoter deletion. These findings indicate that germline BRCA1 promoter deletions are a rare and yet significant mutation event and that they could arise via a novel genetic mechanism. PMID- 11933199 TI - Evaluation and application of denaturing HPLC for mutation detection in Marfan syndrome: Identification of 20 novel mutations and two novel polymorphisms in the FBN1 gene. AB - Mutations in the human fibrillin 1 gene (FBN1) cause the Marfan syndrome (MFS), an autosomal dominant connective tissue disorder. Knowledge about FBN1 mutations is important for early diagnosis, management, and genetic counseling. However, mutation detection in FBN1 is a challenge because the gene is very large in size ( approximately 200 kb) and the approximately 350 mutations detected so far are scattered over 65 exons. Conventional methods for large-scale detection of mutations are expensive, technically demanding, or time consuming. Recently, a high-capacity low-cost mutation detection method was introduced based on denaturing high-performance liquid chromatography (DHPLC). To assess the sensitivity and specificity of this method, we blindly screened 64 DNA samples of known FBN1 genotype exon-by-exon using exon-specific DHPLC conditions. Analysis of 682 PCR amplicons correctly identified 62 out of 64 known sequence variants. In three MFS patients of unknown FBN1 genotype, we detected two mutations and eight polymorphisms. Overall, 20 mutations and two polymorphisms are described here for the first time. Our results demonstrate 1) that DHPLC is a highly sensitive (89-99%, P = 0.05) method for FBN1 mutation detection; but 2) that chromatograms with moderate and weak pattern abnormalities also show false positive signals (in all 45-59%, P = 0.05); 3) that the difference in the chromatograms of heterozygous and homozygous amplicons is mostly independent of the type of sequence change; and 4) that DHPLC column conditions, additional base changes, and the amounts of injected PCR products influence significantly the shape of chromatograms. A strategy for FBN1 mutation screening is discussed. PMID- 11933201 TI - A common frameshift mutation and other variants in GJB4 (connexin 30.3): Analysis of hearing impairment families. AB - Mutations in GJB1, GJB2, GJB3 and GJB6 are involved in hearing impairment. GJB2, GJB3 and GJB6 are also mutated in patients with hyperproliferative skin disorders. The human GJB4 gene has been deduced in silico and a mutation in a family with erythrokeratodermia variabilis has been reported. We describe here the analysis of the GJB4 gene in hearing impairment patients and control subjects. We have identified a common (4%) frameshift mutation (154del4) in GJB4 in both affected and hearing subjects, one patient being homozygous for the mutation. We have also detected five amino acid variants (R103C, R124Q, R160C, C169W and E204A) in individuals that have not skin disorders. While mutation 154del4 is not associated with hearing impairment the involvement of some of the amino acid variants detected here is uncertain. These GJB4 variants should help to define the putative role of connexin 30.3 in both skin disorders and hearing impairment. PMID- 11933202 TI - The identification of eight novel glucocerebrosidase (GBA) mutations in patients with Gaucher disease. AB - Mutations in the gene encoding for the lysosomal enzyme glucocerebrosidase (GBA) result in Gaucher disease. In this study, seven novel missense mutations in the glucocerebrosidase gene (A136E, H162P, K198E, Y205C, F251L, Q350X and I402F) and a splice site mutation (IVS10+2T-->A) were identified by direct sequencing of three amplified segments of the glucocerebrosidase gene. Five of the novel mutations were found in patients with neuronopathic forms of Gaucher disease, two of which, K198E and F251L, appear to be associated with type 2 Gaucher disease. PMID- 11933204 TI - Sequence variations in the mu-opioid receptor gene (OPRM1) associated with human addiction to heroin. AB - Human mu-opioid receptor (OPRM1) is the major site for the analgesic action of most opioid drugs such as morphine, methadone and heroin. It was previously reported that a single nucleotide polymorphism (SNP) in exon1 (c.118A-->G) of OPRM1 might modestly alter the affinity in beta-endorphin-Mu interaction. Using denaturing high performance liquid chromatography (DHPLC) the complete coding region of the OPRM1 gene was screened for SNPs in Han-Chinese heroin addicts and normal control. Three novel SNPs were detected, one in exon3, one in intron3 and one in the 3' untranslated region. The SNP c.118A-->G reportedly altered the interaction of Mu receptor with opioid had no statistically significant correlation with heroin addition in Han Chinese. However, addicted subjects with the SNP in intron2 (IVS2 +31G-->A) tended to show much higher heroin intake dosages than those without this SNP. We also observed that individuals carrying both SNP c.118A-->G and IVS2 +31G-->A consumed relatively more drugs compared to other addicts. Thus our study further highlights the importance of studing the various regions of the mu opioid receptor gene, coding as well as non-coding, for genetic markers that may be linked to, or directly contribute to opioid drug seeking behavior. PMID- 11933203 TI - An osteopontin (SPP1) polymorphism is associated with systemic lupus erythematosus. AB - Osteopontin (SPP1) is a soluble ligand with pleomorphic immunologic activities including activation of macrophage chemotaxis, promotion of Th1 responses, and activation of B1 B cells. It has been implicated in the development of murine lupus and is overexpressed in humans with systemic lupus erythematosus (SLE). We examined a polymorphism of osteopontin for an association with lupus in humans in an effort to determine whether there is any evidence that a genetic predisposition to altered osteopontin expression might explain the overexpression seen in human SLE patients. A silent polymorphism (707C>T, rs1126616) of osteopontin was significantly associated with SLE. Additional associations with renal disease and opportunisitic infections were suggested. This is the first phenotypic association with a polymorphic variant of osteopontin. PMID- 11933205 TI - A BRCA1 mutation in Native North American families. AB - Germline mutations in the BRCA1 (MIM 113705) and BRCA2 (MIM 600185) genes have been identified for breast and ovarian cancer families of diverse ethnic backgrounds. To date, there have been no reports of Native North American families with mutations in BRCA1 or BRCA2. Here we report two families of aboriginal descent both with the same BRCA1 alterations (1510insG, 1506A>G). The families represent two aboriginal Canadian tribes (Cree and Ojibwe), although a common ancestral origin is likely. This is the first evidence of a BRCA1 mutation specific to aboriginal peoples of North America. PMID- 11933206 TI - APC germline mutations identified in Czech patients with familial adenomatous polyposis. AB - Familial adenomatous polyposis (FAP) is an autosomal dominantly inherited predisposition to colorectal cancer, which is caused by germline mutations in the adenomatous polyposis coli (APC) gene. The APC mutations have been investigated in 46 Czech unrelated FAP families and 9 suspected FAP families using DGGE analysis and direct DNA sequencing. We found 25 germline APC mutations and identified 11 which were not previously reported. Of the identified mutations, 10 were 1 to 5 bp deletions, four were 1 bp insertions and six were nonsense, all leading to the formation of premature stop codon. In addition, we detected two mutations in the splice-donor region of APC intron 11, one missense and two samesense mutations. Phenotypes of patients with known and novel types of mutations are presented and discussed. PMID- 11933207 TI - Five novel mutations in the C1 inhibitor gene (C1NH) leading to a premature stop codon in patients with type I hereditary angioedema. AB - Hereditary angioedema (HAE) is a disorder characterised by recurrent attacks of localized subcutaneous or submucosal edema. It is inherited in an autosomal dominant fashion and caused by a deficiency of C1 inhibitor (C1 inh, or C1NH). Most patients with HAE have an absolute deficiency of C1 inh (type I HAE) while the rest (15% of kindreds) synthetize a dysfunctional C1 inh protein (type II HAE). In this report a novel use of denaturing gradient gel electrophoresis (DGGE) followed by direct sequencing of the C1 inhibitor gene is presented. Five novel mutations, one nonsense (p.S48X) and four small deletions resulting in frameshifts (g.2264-2265delAG, g.2304delC, g.8493-8494delCC and g.16676 16677delTG) have been identified in the C1 inhibitor gene in five families with type I HAE. All of these mutations lead to premature termination of translation and thus can be considered causative of the C1 inh deficiency. Moreover, two previously described mutations in the reactive center of C1 inh, p.R444C and p.R444H, have been detected in four unrelated patients with type II HAE. PMID- 11933208 TI - Four novel variants in MC1R in red-haired South African individuals of European descent: S83P, Y152X, A171D, P256S. AB - Skin, hair and eye pigmentation is a polygenic multifactorial trait determined by the cumulative effects of multiple genetic variants and environmental factors. MC1R is one of the genes involved in pigmentation, and has been implicated in the red hair and pale skin trait in human Caucasoid individuals. This study was undertaken to investigate variants in the MC1R gene in Caucasoid individuals in South Africa, who are of European decent. Seven unrelated individuals were studied, all of whom were found to be either homozygous for a single mutation or compound heterozygous for two different mutations. We report four novel MC1R missense mutations: S83P, Y152X, A171D and P256S. This study supports the view that two mutations are necessary, but not necessarily sufficient, to give rise to the red hair and pale skin phenotype. PMID- 11933209 TI - A review of the phenotypic variation due to the Denys-Drash syndrome-associated germline WT1 mutation R362X. AB - The gene WT1 is required for the normal development and function of the urogenital tract. Constitutional mutations are associated with familial Wilms tumor and syndromes such as Denys-Drash syndrome (DDS) characterized by nephropathy, genital anomalies and often a predisposition to Wilms tumor. We report a case of constitutional WT1 mutation in an XX female with multifocal Wilms tumor but no genital anomalies or renal dysfunction and, for the first time, review patients previously reported with this germline mutation. The mutation (1084C>T) changes the amino acid arginine at position 362 to the translation stop codon TGA (R362X) resulting in a predicted truncated protein lacking three of the four zinc finger domains necessary for correct functioning of the gene. This constitutional mutation has been reported to cause a variety of phenotypes in eleven different patients, including the classical Denys-Drash phenotype of diffuse mesangial sclerosis which leads to early renal failure, genital anomalies in XY individuals and Wilms tumors. The absence of mesangial sclerosis and renal failure in our patient excludes DDS. Our case differs from those previously described as the normal kidney tissue shows some small subcapsular glomeruli indicating that the WT1 mutation has impaired nephron development. This patient extends the range and variation of phenotypes that may arise from a specific germline mutation in WT1. PMID- 11933210 TI - Molecular basis of familial hypercholesterolemia in Brazil: Identification of seven novel LDLR gene mutations. AB - Low-density lipoprotein receptor (LDLR) gene mutations cause familial hypercholesterol-emia (FH), one of the most common single gene disorders. The spectrum of LDLR mutations in Brazil is not known. The aim of this study was the characterization of LDLR mutations in 35 unrelated Brazilian patients with heterozygous FH. The promoter region, the 18 exons and the flanking intron sequences of the LDLR gene were screened by PCR-SSCP analysis and by DNA sequencing. In addition, we have screened the apolipoprotein B gene (APOB) for known mutations (R3500Q and R3531C) that cause Familial defective apo B-100 (FDB) by PCR-RFLP procedure. We found two nonsense (E92X and C371X) and six missense LDLR mutations (R236W, G322S, G352D, A370T, C675W and C677Y), that were previously described in FH patients from other populations. We also found five novel missense [G(-20)R, T476P, V503G, D580H and S652R] and two novel frame shift LDLR mutations (FsR757 and FsS828). Four patients were found to carry two different mutations in the LDLR gene: G352D and A370T (one patient), S652R and C675W (one patient) and T476P and V503G (two patients). APOB mutations were not found. These findings demonstrate that there is a broad spectrum of mutations in the LDLR gene in FH individuals from Brazil. PMID- 11933212 TI - Globin gene transfer for the treatment of severe hemoglobinopathies: a paradigm for stem cell-based gene therapy. AB - The prospect of treating blood disorders with genetically modified stem cells is highly promising. This therapeutic approach, however, raises a number of fundamental biological questions, spanning several research fields. Further investigation is required to better understand how to isolate and efficiently transduce hematopoietic stem cells (HSCs), while preserving optimal homing and self-renewing properties; how to design safe vectors permitting controlled expression of the transgene products; and how to promote host repopulation by engrafted HSCs. This article addresses basic issues in stem cell-based gene therapy from the perspective of regulating transgene expression, taking globin gene transfer for the treatment of severe hemoglobinopathies as a paradigm. PMID- 11933213 TI - Oncoretroviral gene transfer to NOD/SCID repopulating cells using three different viral envelopes. AB - BACKGROUND: The aim of this study was to investigate gene transfer to human umbilical cord blood (CB) CD34(+)/CD38(low) and NOD/SCID repopulating cells using oncoretroviral vectors and to compare the transduction efficiency using three different viral envelopes. METHODS: CB cells were transduced on Retronectin using an MSCV-based vector with the gene for GFP (MGIN), which was packaged into three different cell lines giving different envelopes: PG13-MGIN (GALV), 293GPG-MGIN (VSV-G) or AM12-MGIN (amphotropic). RESULTS: Sorted CD34(+)/CD38(low) cells were efficiently transduced after 3 days of cytokine stimulation and the percentage of GFP-positive cells was 61.8+/-6.6% (PG13-MGIN), 26.9+/-3.5% (293GPG-MGIN), and 39.3+/-4.8% (AM12-MGIN). For transplantation experiments, CD34(+) cells were pre stimulated for 2 days before transduction on Retronectin preloaded with vector and with the addition of 1/10th volume of viral supernatant on day 3. On day 4, the expanded equivalent of 2.5x10(5) cells was injected into irradiated NOD/SCID mice. All three pseudotypes transduced NOD/SCID repopulating cells (SRCs) equally well in the presence of serum, but engraftment was reduced when compared with freshly thawed cells. Simultaneous transduction with all three vector pseudotypes increased the gene transfer efficiency to SRCs but engraftment was significantly impaired. There were difficulties in producing amphotropic vectors at high titers in serum-free medium and transduction of CD34(+) cells using VSV-G-pseudotyped vectors under serum-free conditions was very inefficient. In contrast, transduction with PG13-MGIN under serum-free conditions resulted in the maintenance of SRCs during transduction, high levels of engraftment (29.3+/ 6.6%), and efficient gene transfer to SRCs (46.2+/-4.8%). CONCLUSIONS: The best conditions for transduction and engraftment of CB SRCs were obtained with GALV pseudotyped vectors using serum-free conditions. PMID- 11933214 TI - Highly efficient gene transfer into antigen-specific primary mouse lymphocytes with replication-deficient retrovirus expressing the 10A1 envelope protein. AB - BACKGROUND: Introduction of recombinant genes in the genome of primary lymphocytes by virtue of a replication-deficient retrovirus can be used in immunological studies and for cell-based gene therapy. METHODS: Packaging cells GP+E86 producing replication-deficient retrovirus incorporating the genes of enhanced green fluorescent protein (eGFP), C2gamma or C2xi, were generated by calcium phosphate-mediated transfection. Clones with the highest titres of retrovirus vector were isolated from them and their supernatants were used for transduction of PT67 cells. Primary mouse lymphocytes and T-cell hybridoma MD.45 were transduced by centrifugation with retroviral stock. The retroviral content of packaging cell supernatants was determined by dot blotting and hybridization with a DNA probe. RESULTS: PT67 cells produced approximately 50 times more retrovirus vector than the original GP+E86 clones. When retroviral stocks of PT67 and GP+E86 cells were used at 1/50 dilution and undiluted, respectively (to normalize them for retroviral RNA content), the transduction efficiency of mouse T-cell hybridoma was 40% and 5%, respectively. Centrifugation of target cells with retroviral stock at 2000 g for 60 min increased the percentage of transduced cells two- to three-fold. Within a population of cells isolated from the draining lymph nodes of an immunized mouse and reactivated with an antigen, up to 60% of CD4(+) T cells and up to 80% of B cells could be transduced with a transgene in replication-deficient retrovirus packaged by PT67 cells using the optimized gene transfer protocol. CONCLUSIONS: This protocol allows for the generation of packaging cells producing high titres of retrovirus vector. The 10A1 envelope protein is superior to the ecotropic one for the transduction of mouse lymphocytes. PMID- 11933215 TI - Intratumoral activation of cyclophosphamide by retroviral transfer of the cytochrome P450 2B1 in a pancreatic tumor model. Combination with the HSVtk/GCV system. AB - BACKGROUND: Pancreatic cancer is one of the most aggressive human tumors and the development of new therapeutic approaches is particularly urgent since current therapies are not effective. The use of pro-drug-activating genes is a possible approach for cancer gene therapy. METHODS: The present study evaluated the efficiency of the cytochrome P4502B1 (CYP2B1) suicide gene that encodes the enzyme responsible for activating the pro-drug cyclophosphamide (CPA), in pancreatic tumor cells invitro and in vivo. The effects on tumor growth of the combination of two suicide systems, CYP2B1/CPA and herpes simplex virus thymidine kinase gene/ganciclovir (HSVtk/GCV), were also studied. RESULTS: Retroviral CYP2B1 transfer followed by CPA treatment highly sensitized pancreatic tumor cells NP-9, NP-18, and NP-31, and led to stabilization of tumor growth in a pancreatic tumor model. Differences in tumor volume at the end of the treatment were statistically significant when compared with animals injected with CPA alone. The combination of both suicide systems CYP2B1/CPA and HSVtk/GCV in vitro resulted in a potentiation of the killing effect. However, no potentiation was achieved in vivo, although retardation in tumor growth was evident. CONCLUSIONS: The results show that in situ transduction of pancreatic tumor cells with the CYP2B1 gene by retroviral vectors clearly increases the sensitivity to CPA. Moreover, they suggest that in order to achieve a potentiation on cell killing when the two suicide systems HSVtk/GCV and CYP2B1/CPA are combined, co-expression of both genes in the same tumor cell would be necessary. PMID- 11933217 TI - High efficiency lentiviral gene delivery in non-dividing cells by deoxynucleoside treatment. AB - BACKGROUND: Gene therapy has recently been advanced by the development of HIV based vectors that are able to transduce some non-dividing cells. The manipulation of most non-dividing cells remains, however, scarcely efficient. One of the biological mechanisms postulated to prevent powerful transduction of quiescent cells by lentiviral vectors is the paucity of deoxynucleotides (dNTPs). In this study, a novel delivery strategy is developed to improve significantly the efficiency of HIV-based vectors in transducing non-dividing cells. This approach is based on increasing the intracellular availability of dNTPs by incubating target cells with the dNTP precursors, deoxynucleosides (dNSs). METHODS: Mature human monocyte-derived macrophages (14-21 days old) were transduced at a low multiplicity of infection (MOI) of HIV vectors carrying a reporter gene. dNSs were added to the medium during transduction (5 mM dNS) and immediately before post-transduction culture (2.5 mM dNS). Macrophages were harvested 2-7 days after transduction and assayed for transgene expression by cytofluorimetry. RESULTS: The addition of dNS to the medium significantly enhanced the efficiency of transduction of human macrophages by HIV-based vectors. The percentage of cells expressing the transgene rose up to 50% in the presence of dNS, increasing the basal transduction levels up to 35-fold (average=10.8-fold). Furthermore, treatment with dNTP precursors compensated for the wide inter-donor variability, allowing the highest enhancement effects in donors with the lowest basal transduction efficiencies. CONCLUSIONS: This is the first demonstration that a single treatment of non-dividing target cells with exogenous dNS can enhance the efficiency of lentiviral-mediated transduction of cells, allowing for high efficiency gene transfer. The effects of dNTP precursors compensated for both the poor basal levels and the wide inter-donor variability, two major limitations for the transduction of non-dividing cells. Macrophages are a representative model of cells whose permissiveness to gene delivery was increased up to levels suitable for genetic manipulation applications. This simple approach might be transferred to a broader range of quiescent cell types that are scarcely susceptible to lentiviral-based gene delivery due to low dNTP levels. PMID- 11933216 TI - Encapsulated cells producing retroviral vectors for in vivo gene transfer. AB - BACKGROUND: Because gene therapy of the future will primarily take an in vivo approach, a number of problems associated with its current implementation exist. Currently, repeated delivery of a vector in vivo is necessary to ensure adequate transfer of the therapeutic gene. This may lead to the development of an immune response against the vector, thus interfering with gene delivery. To circumvent this problem, retroviral vector packaging cells that permanently produce recombinant retroviral vector particles have been encapsulated. METHODS: Vector (pBAG)-producing amphotropic cells were encapsulated in beads composed of polymerized cellulose sulphate. These capsules were analysed in vitro for expression of the vector construct using X-gal staining, as well as for the release of particles by performing RT-PCR from culture supernatant. Infectivity studies were performed in vitro and in vivo. The latter was assayed using histological sections of the microcapsule and the surrounding area stained for beta-galactosidase activity and by RT-PCR. RESULTS: In culture, the virus producing cells inside the capsules remained viable and released virus into the culture medium for at least 6 weeks. To test whether these capsules, upon implantation into mice, also release vector virions that infect the surrounding cells, two different models were used. In the first, capsules were implanted in the fat pad of the mammary gland of Balb/c mice. The capsules were well tolerated for at least 6 weeks and a self-limiting inflammatory reaction without any other gross immune response was observed during this period. Furthermore, the virus producing cells remained viable. In the second model, SCID mice were immunologically reconstituted by subcutaneous implantation of thymus lobes from MHC-identical Balb/c newborn mice and gene transfer into lymphoid cells was achieved by retroviral vectors released by co-implanted capsules. CONCLUSION: The implantation of such capsules containing cells that continually produce retroviral vector particles may be of use for in vivo gene therapy strategies. The data presented demonstrate the feasibility of the concept. PMID- 11933220 TI - Functional correction of episomal mutations with short DNA fragments and RNA-DNA oligonucleotides. AB - BACKGROUND: Gene correction is an alternative approach to replacement gene therapy. By correcting mutations within the genome, some of the barriers to effective gene therapy are avoided. Homologous nucleic acid sequences can correct mutations by inducing recombination or mismatch repair. Recently, encouraging data have been presented using both short DNA fragments (SDFs) and RNA-DNA oligonucleotides (RDOs) in experimental strategies to realize clinical gene correction. METHODS: The delivery of labelled SDFs and RDOs to a variety of cell lines was tested using both FACS analysis and confocal microscopy. A GFP-based reporter system was constructed, containing a nonsense mutation, to allow quantitation of gene correction in living cells. This reporter was used to compare efficiencies of functional gene correction using SDFs and RDOs in arange of mammalian cell lines. RESULTS: The delivery experiments highlight the inefficient delivery of SDFs and RDOs to the nucleus using polyethylenimine (PEI) transfection. This study compared the episomal correction efficiency of the reporter plasmid mediated by SDFs and RDOs within different cell types; low levels of functional correction were detected in cell culture. CONCLUSIONS: Whilst delivery of PEI-complexed SDFs or RDOs to the cell is highly effective, nuclear entry appears to be a limiting factor. SDFs elicited episomal GFP correction across a range of cell lines, whereas RDOs only corrected the reporter in a cell line that overexpresses RAD51. PMID- 11933219 TI - Receptor-mediated delivery of an antisense gene to human brain cancer cells. AB - BACKGROUND: The goal of this work was the development of a gene targeting technology that will enable the delivery of therapeutic genes to brain cancer cells in vivo following intravenous administration. High-grade brain gliomas overexpress the epidermal growth factor receptor (EGFR) and EGFR antisense gene therapy could reduce the growth of EGFR-dependent gliomas. METHODS: A human EGFR antisense gene driven by the SV40 promoter in a non-viral plasmid carrying elements that facilitate extra-chromosomal replication was packaged in the interior of 85 nm pegylated immunoliposomes (PILs). The PILs were targeted to U87 human glioma cells with the 83-14 murine monoclonal antibody (MAb) to the human insulin receptor (HIR). RESULTS: Confocal fluorescent microscopy demonstrated that the unconjugated HIR MAb is rapidly internalized by the glioma cells. Endocytosis followed by entry into the nucleus was also demonstrated for the HIR MAb conjugated PILs carrying fluorescein-labeled plasmid DNA. The PILs delivered exogenous genes to virtually all cells in culture, based on beta-galactosidase histochemistry. The targeting of a luciferase gene to the U87 cells with the PILs resulted in luciferase levels in excess of 150 pg/mg protein after 72 h of incubation. The level of luciferase gene expression in the U87 cells achieved with the PIL gene targeting system was comparable to that with lipofectamine. Targeting the EGFR antisense gene to U87 glioma cells with the PILs resulted in more than 70% reduction in [(3)H]thymidine incorporation into the cells; this was paralleled by a 79% reduction in the level of immunoreactive EGFR. CONCLUSION: The present work describes the targeting of an EGFR antisense gene to human brain cancer cells, which results in a 70-80% inhibition in cancer cell growth. PILs provide a new approach to gene targeting that is effective in vivo following intravenous administration without viral vectors. PMID- 11933218 TI - Polyethyleneimine-based immunopolyplex for targeted gene transfer in human lymphoma cell lines. AB - BACKGROUND: Specific and efficient delivery of genes into targeted cells is a priority objective in non-viral gene therapy. Polyethyleneimine-based polyplexes have been reported to be good non-viral transfection reagents. However, polyplex mediated DNA delivery occurs through a non-specific mechanism. This article reports the construction of an immunopolyplex, a targeted non-viral vector based on a polyplex backbone, and its application in gene transfer over human lymphoma cell lines. METHODS: Targeting elements (biotin-labeled antibodies), which should recognize a specific element of the target cell membrane and promote nucleic acid entry into the cell, were attached to the polyplex backbone through a bridge protein (streptavidin). Immunopolyplex transfection activity was studied in several hematological cell lines [Jurkat (CD3+/CD19-), Granta 519 (CD3-/ CD19+), and J.RT3-T3.5 (CD3-/CD19-)] using the EGFP gene as a reporter gene and anti-CD3 and anti-CD19 antibodies as targeting elements. Transfection activity was evaluated via green fluorescence per cell and the percentage of positive cells determined by flow cytometry. RESULTS: A significant selectivity of gene delivery was observed, since the anti-CD3 immunopolyplex worked only in Jurkat cells while the anti-CD19 immunopolyplex worked only in the Granta cell line. Moreover, transfection of a CD3+/CD3- cell mixture with anti-CD3 immunopolyplexes showed up to 16-fold more transfection in CD3+ than in CD3- cells. Several non-specific transfection reagents showed poor or no transfection activity. CONCLUSION: It is concluded that immunopolyplex is a good non-viral vector for specific and selective nucleic acid delivery. Immunopolyplex design allows easy replacement of the targeting element (antibody) - the streptavidin-polyplex backbone remaining intact - thereby conferring high versatility. PMID- 11933221 TI - The use of low-molecular-weight PEIs as gene carriers in the monkey fibroblastoma and rabbit smooth muscle cell cultures. AB - BACKGROUND: Polyethylenimines (PEIs) and cationic polymers have been used successfully in gene delivery. In earlier reports, only large PEIs (MW>10 000) have shown significant transfection efficiency. In the present study, the roles of small PEIs (MW 700 and 2000) were studied as additional compounds to see if they can improve gene delivery with cationic liposomes. METHODS: The TKBPVlacZ expression plasmid was transfected in the CV1-P (monkey fibroblastoma) and SMC (rabbit smooth muscle) cell lines using various combinations of PEIs (MW 700, 2000, and 25 000) and Dosper liposomes. The transfection efficiency was determined with the fluorometric ONPG (o-nitrophenol-beta-D-galactopyranoside) assay and histochemical X-gal staining. The toxicity of the transfection reagents was estimated by the MTT [3-(4,5-dimethylthiazolyl-2)-2,5-diphenyl tetrazolium bromide] assay. RESULTS: Transfection of TKBPVlacZ plasmid by the small PEIs (MW 700 and 2000) combined with Dosper liposomes was associated with high expression of the lacZ reporter gene in the CV1-P and SMC cell lines. The transfection efficiencies of the low-molecular-weight PEI/liposome combinations were several fold higher than those of PEIs or liposomes alone. PEI/liposome combinations had no toxicity on the cell lines tested. CONCLUSIONS: The low-molecular-weight PEIs could be used successfully for gene delivery when combined with the cationic liposomes, resulting in a synergistic increase of the transfection efficiency in both cell lines studied. PMID- 11933222 TI - Therapeutic levels of human factor VIII in mice implanted with encapsulated cells: potential for gene therapy of haemophilia A. AB - BACKGROUND: A gene therapy delivery system based on microcapsules enclosing recombinant cells engineered to secrete a therapeutic protein has been evaluated. The microcapsules are implanted intraperitoneally. In order to prevent cell immune rejection, cells are enclosed in non-antigenic biocompatible alginate microcapsules prior to their implantation into mice. It has been shown that encapsulated myoblasts can deliver therapeutic levels of Factor IX (FIX) in mice. The delivery of human Factor VIII (hFVIII) in mice using microcapsules was evaluated in this study. METHODS: Mouse C2C12 myoblasts and canine MDCK epithelial kidney cells were transduced with MFG-FVIII (B-domain deleted) vector. Selected recombinant clones were enclosed in alginate microcapsules. Encapsulated recombinant clones were subsequently implanted intraperitoneally into C57BL/6 and immunodeficient SCID mice. RESULTS: Plasma of mice receiving C2C12 and encapsulated MDCK cells had transient therapeutic levels of FVIII in immunocompetent C57BL/6 mice (up to 20% and 7% of physiological levels, respectively). In addition, FVIII delivery in SCID mice was also transient, suggesting that a non-immune mechanism must have contributed to the decline of hFVIII in plasma. Quantitative RT-PCR analysis confirmed directly that the decline of hFVIII is due to a reduction in steady-state hFVIII mRNA, consistent with transcriptional repression. Furthermore, encapsulated cells retrieved from implanted mice were viable, but secreted FVIII ex vivo at three-fold lower levels than the pre-implantation levels. In addition, antibodies to hFVIII were detected in immunocompetent C57BL/6 mice. CONCLUSIONS: Implantable microcapsules can deliver therapeutic levels of FVIII in mice, suggesting the potential of this gene therapy approach for haemophilia A. The findings suggest vector down regulation in vivo. PMID- 11933229 TI - Vancomycin resistance: small molecule approaches targeting the bacterial cell wall biosynthesis. PMID- 11933228 TI - Beyond Watson and Crick: DNA methylation and molecular enzymology of DNA methyltransferases. AB - DNA methyltransferases catalyze the transfer of a methyl group from S-adenosyl-L methionine to cytosine or adenine bases in DNA. These enzymes challenge the Watson/Crick dogma in two instances: 1) They attach inheritable information to the DNA that is not encoded in the nucleotide sequence. This so-called epigenetic information has many important biological functions. In prokaryotes, DNA methylation is used to coordinate DNA replication and the cell cycle, to direct postreplicative mismatch repair, and to distinguish self and nonself DNA. In eukaryotes, DNA methylation contributes to the control of gene expression, the protection of the genome against selfish DNA, maintenance of genome integrity, parental imprinting, X-chromosome inactivation in mammals, and regulation of development. 2) The enzymatic mechanism of DNA methyltransferases is unusual, because these enzymes flip their target base out of the DNA helix and, thereby, locally disrupt the B-DNA helix. This review describes the biological functions of DNA methylation in bacteria, fungi, plants, and mammals. In addition, the structures and mechanisms of the DNA methyltransferases, which enable them to specifically recognize their DNA targets and to induce such large conformational changes of the DNA, are discussed. PMID- 11933230 TI - NMR solution structure, backbone mobility, and homology modeling of c-type cytochromes from gram-positive bacteria. AB - The solution structure of oxidized cytochrome c(553) (71 amino acid residues) from the Gram-positive bacterium Bacillus pasteurii is here reported and compared with the available crystal structure. The solution structure is obtained from 1609 meaningful NOE data (22.7 per residue), 76 dihedral angles, and 59 pseudocontact shifts. The root mean square deviations from the average structure are 0.25+/-0.07 and 0.59+/-0.13 A for the backbone and all heavy atoms, respectively, and the quality assessment of the structure is satisfactory. The solution structure closely reproduces the fold observed in the crystal structure. The backbone mobility was then investigated through amide (15)N relaxation rate and (15)N-(1)H NOE measurements. The protein is rigid in both the sub-nanosecond and millisecond time scales, probably due to the relatively large heme:number of amino acids ratio. Modeling of eight c-type cytochromes from other Gram-positive bacteria with a high sequence identity (>30 %) to the present cytochrome c(553) was performed. Analysis of consensus features accounts for the relatively low reduction potential as being due to extensive heme hydration and indicates residues 34-35, 44-46, 69-72, and 75 as a conserved hydrophobic patch for the interaction with a protein partner. At variance with mitochondrial c-type cytochrome, this protein does not experience pH-dependent coordination equilibria. The reasons for this difference are analyzed. PMID- 11933231 TI - Ratcheting up vir gene expression in Agrobacterium tumefaciens: coiled coils in histidine kinase signal transduction. AB - The transmembrane histidine kinase VirA is responsible for the recognition of information from several plant-derived xenognostic signals that control gene transfer between Agrobacterium tumefaciens and its eukaryotic host. As with other histidine autokinases, VirA appears to exist as a homodimer within the inner membrane of the bacterium. In this study, we identify the putative homodimeric coiled-coil-like motifs Helix TM2 (amino acids (aa) 259-288) and Helix C (aa 293 327) within the previously assigned signal input domain. The functional importance of these coiled-coil interactions in signal-mediated VirA activation is investigated by the construction of fusion proteins with the leucine zipper domain of the transcription factor GCN4. Replacement of the membrane-spanning and periplasmic domains of VirA with the GCN4 leucine zipper gave functional proteins with increased signal-induced vir gene expression. When the GCN4 fusion was used to conformationally bias the interface of the Helix C coiled coil, constitutively active chimeras were created. The activity of these constructs was dependent on the interface of the Helix C coiled coil, and a ratchet model is proposed in which VirA activation is achieved by signal-induced switching of the interfaces of the homodimer. Since VirA functions as a transducer and integrates various host cues indirectly, these data highlight its role as an "antenna" for the tumor inducing (Ti) plasmid, able to monitor the host proteome so as to select for successful xenognostic signaling strategies. PMID- 11933232 TI - A new family of beta-hairpin mimetics based on a trypsin inhibitor from sunflower seeds. AB - The ability of proteases to regulate many aspects of cell function and defense accounts for the considerable interest in the design of novel protease inhibitors. There are many naturally occurring proteinaceous serine protease inhibitors, one of which is a 14 amino acid cyclic peptide from sunflower seeds that shows both sequence and conformational similarity with the trypsin-reactive loop of the Bowman-Birk family of serine protease inhibitors. This inhibitor adopts a beta-hairpin conformation when bound at the active site of bovine beta trypsin. We illustrate here an approach to inhibitor design in which the beta hairpin from the naturally occurring peptide is transplanted onto a hairpin inducing template. Two mimetics with the sequences RC*TKSIPPIC*F (where C*C* is a disulfide) and TKSIPPI are studied, each mounted onto a D-Pro-L-Pro template. NMR studies revealed a well-defined beta-hairpin conformation for each mimetic in aqueous solution; this conformation is closely related to the trypsin-bound conformation of the natural inhibitor and includes a cis-Ile-Pro peptide bond. Both mimetics inhibit trypsin in the mid nanomolar range. An alanine scan revealed the importance for inhibitory activity of the specificity-determining Lys residue and of the first but not the second Pro residue in the IPPI motif. Since these hairpin mimetics can be prepared by parallel combinatorial synthesis, this family of molecules may be a useful starting point for the discovery of other biologically or medicinally useful serine protease inhibitors. PMID- 11933233 TI - Formation of DNA triple helices by an oligonucleotide conjugated to a fluorescent ruthenium complex. AB - A conjugate of a triple helix forming oligonucleotide (TFO) and the Lambda and Delta enantiomers of the ruthenium diphenanthroline dipyridophenazine complex [Ru(phen)(2)dppz](2+) was synthesized. The ruthenium complex was attached to the 5'-end of the TFO through the dppz moiety. This conjugate formed a stable triple helix with the polypurine tract (PPT) sequence from HIV proviral DNA. The thermal denaturation temperature of the triplex was increased by 12 degrees C. One remarkable property of the Delta-[Ru(phen)(2)dppz](2+) complex is a strong increase in its fluorescence when it intercalates into DNA. While the fluorescence of the oligonucleotide conjugate was very weak, the formation of a duplex with a complementary sequence or of a triple helix with a target duplex resulted in a large increase in fluorescence of the Delta enantiomer. The increase in fluorescence allowed us to follow the kinetics of duplex and triplex formation by fluorescence spectrometry. In contrast, the Lambda enantiomer gave a much smaller fluorescence change when a triplex was formed, even though the stability of the triplex was comparable to that of the Delta enantiomer. The property was ascribed to intercalation of the dipyridophenazine moiety of the Delta enantiomer into DNA and subsequent threading of the ruthenium complex through the DNA double helix. Salt effects were consistent with the involvement of DNA breathing in the formation of the intercalating complex. PMID- 11933234 TI - Moenomycin-mediated affinity purification of penicillin-binding protein 1b. AB - The antibiotic moenomycin A inhibits the biosynthesis of peptidoglycan, the main structural polymer of the bacterial cell wall. The inhibition is based on a reversible binding of the antibiotic to one of the substrate binding sites at enzymes such as the penicillin binding protein 1b (PBP 1b). This binding has been employed to isolate PBP 1b by affinity chromatography. Suitable ligands have been prepared from moenomycin A and coupled both to affinity supports and to surface plasmon resonance sensor surfaces. The reactions that take place upon immobilization of the ligands to the affinity support and the sensor surface, respectively, have been studied in detail. With the help of surface plasmon resonance the optimal conditions for binding of PBP 1b to moenomycin-derivated ligands have been established. For the first time the selective binding of the moenomycin sugar moiety to the enzyme has been demonstrated. PMID- 11933235 TI - Synthesis and evaluation of organosilicon inhibitors of active purine transport in human osteoblasts. AB - In the search for new compounds that might, once incorporated into biomaterials, stimulate the natural processes of bone regeneration, a new series of silicon containing alkyl nucleobase analogues has been synthesized. An active hypoxanthine transport process in human osteoblasts was demonstrated, with an apparent Michaelis constant of 2.3 microM and a maximum possible rate of 0.47 pmol s(-1) x 10(6) cell. The synthesized analogues were tested for toxicity in human osteoblasts. Nontoxic analogues were tested in competition transport studies with [(14)C]hypoxanthine. Two of them were found to inhibit the active transport of hypoxanthine in human osteoblasts, with IC(50) values of 6.5 and 11.6 microM. PMID- 11933236 TI - Combined biosynthetic pathway for de novo production of UDP-galactose: catalysis with multiple enzymes immobilized on agarose beads. AB - Regeneration of sugar nucleotides is a critical step in the biosynthetic pathway for the formation of oligosaccharides. To alleviate the difficulties in the production of sugar nucleotides, we have developed a method to produce uridine diphosphate galactose (UDP-galactose). The combined biosynthetic pathway, which involves seven enzymes, is composed of three parts: i) the main pathway to form UDP-galactose from galactose, with the enzymes galactokinase, galactose-1 phosphate uridyltransferase, UDP-glucose pyrophosphorylase, and inorganic pyrophosphatase, ii) the uridine triphosphate supply pathway catalyzed by uridine monophosphate (UMP) kinase and nucleotide diphosphate kinase, and iii) the adenosine triphosphate (ATP) regeneration pathway catalyzed by polyphosphate kinase with polyphosphate added as an energy resource. All of the enzymes were expressed individually and immobilized through their hexahistidine tags onto nickel agarose beads ("super beads"). The reaction requires a stoichiometric amount of UMP and galactose, and catalytic amounts of ATP and glucose 1 phosphate, all inexpensive starting materials. After continuous circulation of the reaction mixture through the super-bead column for 48 h, 50 % of the UMP was converted into UDP-galactose. The results show that de novo production of UDP galactose on the super-bead column is more efficient than in solution because of the stability of the immobilized enzymes. PMID- 11933237 TI - A novel cyclodextrin-derived tellurium compound with glutathione peroxidase activity. AB - A novel dicyclodextrinyl ditelluride (2-TeCD) compound was devised as a functional mimic of the glutathione peroxidase (GPX) enzymes that normally remove hydroperoxides from the cell. The GPX activity of the mimic was found to be 46.7 U microM(-1), which is 46 times as active as Ebselen, a well-known GPX mimic. A detailed steady-state kinetic study was undertaken to probe the reason for the high catalytic efficiency of 2-TeCD. This high efficiency can be explained based on both the binding of the substrate to the cyclodextrin and the catalytic mechanism of 2-TeCD, which is different from that of diselenide compounds. 2-TeCD exhibits good water solubility and is chemically and biologically stable. The biological effect of 2-TeCD was evaluated by its ability to protect mitochondria from oxidative damage. 2-TeCD exhibited excellent antioxidant capacity in comparison with Ebselen. PMID- 11933240 TI - Editorial. PMID- 11933241 TI - Editorial. PMID- 11933238 TI - Protective activities of some phenolic 1,3-diketones against lipid peroxidation: possible involvement of the 1,3-diketone moiety. AB - The protective activities of four ginger-derived phenolic 1,3-diketones (1-4) and curcumin (5) against lipid peroxidation was studied by using different biologically relevant model systems and pulse radiolysis. The extraordinary activity of 5 vis-a-vis 1-4 against Fe(2+)-mediated peroxidation may be attributed to the additional phenolic hydroxy group in the former, which lends it better iron-chelating and radical-scavenging properties. In iron-independent peroxidation, however, the ginger constituent [6]-dehydrogingerdione (1) showed activity comparable to that of 5; this indicates its higher affinity for the lipid peroxide radical (LOO(.)), due to its higher hydrophobicity. A very high rate constant for the reaction between 1 and Cl(3)COO(.), measured by pulse radiolysis, not only confirmed this, but also established the superior antioxidant efficacy of 1 in comparison to vitamins E and C. This was also evident from the results obtained from a liposomal peroxidation study with 1 and vitamin C. This study also established a synergistic effect of the latter on the antioxidant activity of 1. HPLC analysis of the products of the reaction between 1 and Cl(3)COO(.) revealed the formation of higher concentrations of ferulic acid (7), along with vanillin (6). The presence of ascorbate affected the generation of 7 more than it did that of 6. On this basis, a mechanism for the antioxidant action of 1 has been proposed, which suggests the contribution of the phenolic group as well as the active methylene group of the 1,3-diketones. PMID- 11933242 TI - Transformation of organic molecules on the low-valent [M(Ph(2)PCH(2)CH(2)PPh(2))(2)] moiety derived from trans [M(N(2))(2)(Ph(2)PCH(2)CH(2)PPh(2))(2)] or related complexes (M = Mo, W). AB - A zero-valent [M(Ph(2)PCH(2)CH(2)PPh(2))(2)] moiety (M = Mo, W) generated in situ by dissociation of the N(2) ligands in trans [M(N(2))(2)(Ph(2)PCH(2)CH(2)PPh(2))(2)] can activate pi-accepting organic molecules including isocyanides and nitriles, which undergo the electrophilic attack caused by a strong pi-donation from a zero-valent metal center. Cleavage of a variety of C-X bonds (X = H, C, N, O, P, halogen) also occurs at their electron-rich sites through oxidative addition to form reactive intermediates, which subsequently degradate to yield smaller molecules either bound to or dissociated from the metal center. The mechanism is substantiated unambiguously by isolation of numerous intermediate stages. PMID- 11933243 TI - Chemistry and biology of phototropism-regulating substances in higher plants. AB - Most people are familiar with the sight of a young seedling bending towards a window or the brightest source of light to which it is exposed. This directional growth response is known as phototropism, which is caused by a lateral growth promoting auxin in the bending organ (Cholodny-Went theory, cited in high school textbook). Recently, however, Bruinsma et al., Weiler et al., and Hasegawa et al. independently found that the shaded half did not contain more auxin than the illuminated one. Instead it was found that the even distribution of auxin was accompanied by a lateral gradient of growth inhibiting substances during phototropic curvature (Bruinsma-Hasegawa theory). We have isolated some photo induced growth inhibitory substances related to phototropism, benzoxazolinones from light-grown maize shoots (Zea mays L.), raphanusanins from radish hypocotyl (Raphanus sativus var. hortensis f. gigantissimus M.), indolyacetonitrile from light-grown shoots (Brassica oleacea L.), 8-epixanthatin from sunflower hypocotyl (Helianthus annus L.), and quite recently uridine from oat coleoptile (Avena sativa L.). Chemical analyses have shown phototropic stimulations to cause curvature by inducing a local unequal distribution of growth-inhibiting substances that antagonize auxin in its cell-elongating activity. Finally, a model is presented for further studies on phototropism. PMID- 11933244 TI - Stereospecificity for the hydrogen transfer of pyridoxal enzyme reactions. AB - We have studied the stereospecificities of various pyridoxal 5'-phosphate dependent enzymes for the hydrogen transfer between the C-4' of a bound coenzyme and the C-2 of a substrate in the transamination catalyzed by the enzymes. Prior to our studies, pyridoxal enzymes so far studied were reported to catalyze the hydrogen transfer only on the si-face of the planar imine intermediate formed from substrate and coenzyme. This finding had been considered as the evidence that pyridoxal enzymes have evolved divergently from a common ancestral protein, because identity in the stereospecificity reflects the similarity in the active site structure, in particular in the geometrical relationship between the coenzyme and the active site base participating in the hydrogen transfer. However, we found that D-amino acid aminotransferase, branched-chain L-amino acid aminotransferase, and 4-amino-4-deoxychorismate lyase catalyze the re-face specific hydrogen transfer, and that amino acid racemases catalyze the nonstereospecific hydrogen transfer. These findings suggest the convergent evolution of pyridoxal enzymes. Crystallographical studies have shown that the stereospecificity reflects the active-site structure of the enzymes, and that the enzymes with the same fold exhibit the same stereospecificity. The active site structure with the catalytic base being situated on the specific face of the cofactor has been conserved during the evolution among the pyridoxal enzymes of the same family. PMID- 11933245 TI - Release of enzyme strain during catalysis reduces the activation energy barrier. AB - Several mechanisms have been considered as principal factors in enhancing the catalytic reaction velocity of enzymes: approximation, covalent catalysis, general acid-based catalysis, and strain. Among them, the strain on the substrate and/or the enzyme is often found to be brought about on association of the substrate and the enzyme. If this strain is released in the transition state, it contributes to enhancing the k(cat) value, although it does not change the k(cat)/K(m) value. In aspartate aminotransferase, however, we found by analysis of the Schiff base pK(a) values that the unliganded enzyme carries a strain in the protonated Schiff base formed between the coenzyme pyridoxal phosphate and a lysine residue. This bond is cleaved in most of the reaction intermediates, including the transition state. As a result, the activation energy between the free enzyme plus substrate and the transition state is decreased by 16 kJ/mol, equal to the value of the strain energy. The net effect of this strain is enhancement (10(3)-fold) of the catalytic efficiency in terms of k(cat)/K(m), the more important indicator of the catalytic efficiency at low concentration of the substrate. PMID- 11933246 TI - Microfabricated devices for fluid mixing and their application for chemical synthesis. AB - Over the last few decades, the processes of miniaturization, integration, and automation have revolutionized the world of science and industry. Within a chemical reaction process the unit operations, mixing, heating, and cooling, can be regarded as key steps. In microreactors, enhanced heat and mass transport, due to small characteristic dimensions together with large surface to volume ratios, are expected to open up a whole range of new possibilities. Increase in reaction yield, reduction of reaction time as well as byproduct formation, inherent process safety, and even completely new process routes are some of the advantages associated with microTAS (micro Total Analysis Systems) or microSYNTAS (micro SYNthesis Total Analysis Systems). This article aims to describe the development of microfabricated devices for fluid mixing, so-called micromixers, and their application for chemical synthesis. PMID- 11933247 TI - The development of lithium ion secondary batteries. AB - Lithium ion secondary batteries (LIBs) were successfully developed as battery systems with high volumetric and gravimetric energy densities, which were inherited from lithium secondary batteries (LSBs) with metallic lithium anodes. LSBs have several drawbacks, however, including poor cyclability and quick-charge rejection. The cell reaction in LIB is merely a topochemical one, namely the migration of lithium ions between positive and negative electroces. No chemical changes were observed in the two electrodes or in the electrolytes. This results in little chemical transformation of the active electrode materials and electrolytes, and thus, LIBs can overcome the weaknesses of LSBs; for example, LIBs show excellent cyclability and quick-charge acceptance. Many difficulties, however, were encountered during the course of development, including capacity fade during cycling and safety issues. This article is the story of the development of LIBs and it describes how the difficulties were surmounted. PMID- 11933248 TI - Consecutive carbon-carbon bond formation approach in tandem cyclization reactions. AB - The conventional tandem cyclization reactions involve the formation of alternating carbon-carbon bonds, whereas the newly developed cyclization reactions involve the formation of consecutive carbon-carbon bonds, in which N aziridinylimines have been utilized as geminal radical acceptor and donor equivalents in a single operation. This unprecedented tandem cyclization approach becomes feasible by the successful generation of 5- and 6-membered ring radicals by radical cyclizations of N-aziridinylimines. The same notion can be applied to the anionic cyclizations of N-aziridinylimines, thereby allowing anionic consecutive carbon-carbon bond formation. This approach has great synthetic potential, particularly for the construction of quaternary carbon centers, and it provides highly efficient routes for the synthesis of natural products. PMID- 11933249 TI - Artificial molecular-level machines. AB - The concept of "machine" can be extended to the molecular level by designing supramolecular species capable of performing mechanical-like movements as a consequence of an appropriate energy supply. Molecular-level machines operate via electronic and nuclear rearrangements, for example, through some kind of chemical reaction. Like macroscopic machines, they are characterized by: (i) the kind of energy input supplied to make them work, (ii) the kind of movement performed by their components, (iii) the way in which their operation can be controlled and monitored, (iv) the possibility to repeat the operation at will and establish a cyclic process, (v) the time scale needed to complete a cycle of operation, and (vi) the function performed. A crucial issue is that concerning energy supply. Artificial machines powered by chemical energy ("fuels") produce waste products whose accumulation compromises the operation of the machine unless they are removed from the system. Photochemical and electrochemical energy inputs, however, can be used to make a machine work without formation of waste products. Examples of chemically, electrochemically, and photochemically powered machines investigated in our laboratory are reviewed, and future directions for the construction of novel machines are illustrated. The two most interesting kinds of applications of molecular-level machines are related to the mechanical aspect, which can be exploited, for example, for molecular-level transportation purposes, and the logic aspect, which can be exploited for information processing at the molecular level and, in the long run, for the construction of molecular level (chemical) computers. PMID- 11933250 TI - From cofactor to enzymes. The molecular evolution of pyridoxal-5'-phosphate dependent enzymes. AB - The pyridoxal-5'-phosphate (vitamin B(6))-dependent enzymes that act on amino acid substrates have multiple evolutionary origins. Thus, the common mechanistic features of B(6) enzymes are not accidental historical traits but reflect evolutionary or chemical necessities. The B(6) enzymes belong to four independent evolutionary lineages of paralogous proteins, of which the alpha family (with aspartate aminotransferase as the prototype enzyme) is by far the largest and most diverse. The considerably smaller beta family (tryptophan synthase beta as the prototype enzyme) is structurally and functionally more homogenous. Both the D-alanine aminotransferase family and the alanine racemase family consist of only a few enzymes. The primordial pyridoxal-5'-phosphate-dependent protein catalysts apparently first diverged into reaction-specific protoenzymes, which then diverged further by specializing for substrate specificity. Aminotransferases as well as amino acid decarboxylases are found in two different evolutionary lineages, providing examples of convergent enzyme evolution. The functional specialization of most B(6) enzymes seems to have already occurred in the universal ancestor cell before the divergence of eukaryotes, archebacteria, and eubacteria 1500 million years ago. Pyridoxal-5'-phosphate must have emerged very early in biological evolution; conceivably, metal ions and organic cofactors were the first biological catalysts. To simulate particular steps of molecular evolution, both the substrate and reaction specificity of existent B(6) enzymes were changed by substitution of active-site residues, and monoclonal pyridoxal-5' phosphate-dependent catalytic antibodies were produced with selection criteria that might have been operative in the evolution of protein-assisted pyridoxal catalysis. PMID- 11933251 TI - Nanopore and nanoparticle catalysts. AB - The design, atomic characterization, performance, and relevance to clean technology of two distinct categories of new nanocatalysts are described and interpreted. Exceptional molecular selectivity and high activity are exhibited by these catalysts. The first category consists of extended, crystallographically ordered inorganic solids possessing nanopores (apertures, cages, and channels), the diameters of which fall in the range of about 0.4 to about 1.5 nm, and the second of discrete bimetallic nanoparticles of diameter 1 to 2 nm, distributed more or less uniformly along the inner walls of mesoporous (ca. 3 to 10 nm diameter) silica supports. Using the principles and practices of solid-state and organometallic chemistry and advanced physico-chemical techniques for in situ and ex situ characterization, a variety of powerful new catalysts has been evolved. Apart from those that, inter alia, simulate the behavior of enzymes in their specificity, shape selectivity, regio-selectivity, and ability to function under ambient conditions, many of these new nanocatalysts are also viable as agents for effecting commercially significant processes in a clean, benign, solvent-free, single-step fashion. In particular, a bifunctional, molecular sieve nanopore catalyst is described that converts cyclohexanone in air and ammonia to its oxime and caprolactam, and a bimetallic nanoparticle catalyst that selectively converts cyclic polyenes into desirable intermediates. Nanocatalysts in the first category are especially effective in facilitating highly selective oxidations in air, and those in the second are well suited to effecting rapid and selective hydrogenations of a range of organic compounds. PMID- 11933252 TI - Syntheses of semiconductor nanoparticles using single-molecular precursors. AB - Methods for the preparation of II-VI, III-V, and II-V as well as other compound semiconductor nanoparticles using main group single-molecular precursors have been developed. The work involves the design and synthesis of compounds containing all the elements required within the desired nanoparticulate material. Precursors are tailored to give reproducible, clean decomposition at moderate temperatures, leading to high quality, defect free, mono-dispersed nanoparticles. In this article we cover key aspects of precursor and nanoparticle synthesis. One of the more successful and reproducible series of single-source precursors used, and the one on which we have concentrated our research efforts, is the bis(dialkyldithio-/diseleno-carbamato)cadmium(II)/zinc(II) compounds, M(E(2)CNR(2))(2) (M = Zn or Cd, E = S or Se, and R = alkyl) for the preparation of chalcogenide nanoparticulate materials. Preliminary mechanistic studies suggest that the precursor to nanoparticle deposition route is strongly influenced by the alkyl substituent groups present, and may well determine the phase and quality of the final metal chalcogenide nanoparticles produced. Herein we discuss the synthesis of semiconductor nanoparticles using such single molecular precursors. PMID- 11933253 TI - Bacterial photosynthesis begins with quantum-mechanical coherence. AB - In the antenna system of photosynthetic bacteria, pigments form circular aggregates whose excitations are excitons with quantum-mechanical coherence extending over many pigments. These excitons play crucial roles in light harvesting, storage, and excitation-energy transfer (EET). EET takes place rapidly to and/or from optically forbidden exciton states, without total transition dipole, within the antenna system and to the reaction center. Such EETs cannot be rationalized by Forster's formula, the traditional theory on EET, because it allows EET only between optically allowed states. The coherence in the excitons seems to prohibit rapid EET on this formula. The bacteria overcome this difficulty by circumventing the coherence, using the effects of the physical size of an aggregate that is larger than the shortest distance between pigments in the donor and pigments in the acceptor. The shortest-distance pair therein cannot detect whether the aggregate has a nonvanishing total transition dipole or not, since the pair see effectively only the transition dipole on the other pigment in themselves. The transition dipole facilitates rapid EET even to and/or from optically forbidden exciton states. Such EETs have enabled us to develop a general formula for the rate constant of EET. This is a formula in the weak interaction limit, and so is Forster's formula, but it correctly takes into account the above size effect. PMID- 11933257 TI - Methylenetetrahydrofolate reductase: a common human polymorphism and its biochemical implications. AB - Methlenetetrahydrofolate (CH2-H4folate) is required for the conversion of homocysteine to methionine and of dUMP to dTMP in support of DNA synthesis, and also serves as a major source of one carbon unit for purine biosynthesis. This review presents biochemical studies of a human polymorphism in methylenetetrahydrofolate reductase, which catalyzes the reaction shown below. The mutation decreases the flux of CH2-H4folate into CH3-H4folate, and is associated with both beneficial and deleterious effects that can be traced to the molecular effect of the substitution of alanine 222 by valine. PMID- 11933258 TI - Green organic syntheses: organic carbonates as methylating agents. AB - Dimethylcarbonate (DMC) is a valuable methylating reagent that can replace methyl halides and dimethylsulfate in the methylation of a variety of nucleophiles. It couples tunable reactivity and unprecedented selectivity towards mono-C- and mono N-methylation. In addition, it is a prototype example of a green reagent, because it is nontoxic, is made by a clean process, is biodegradable, and reacts in the presence of a catalytic amount of base, thereby avoiding the formation of undesirable inorganic salts as by-products. Depending on the reaction conditions, DMC can be reacted under plug-flow, CSTR, or batch conditions. Other remarkable reactions are those where DMC behaves as an oxidant. The reactivity of other carbonates is reported as well. PMID- 11933260 TI - Indole synthesis by radical cyclization of o-alkenylphenyl isocyanides and its application to the total synthesis of natural products. AB - Development of indole synthesis by tin-mediated radical cyclization of o alkenylphenyl isocyanide is described. Upon heating o-alkenylphenyl isocyanide in the presence of tri-n-butyltin hydride and AIBN, 2-stannyl-3-substituted indole was formed via 5-exo-trig cyclization of the imidoyl radical intermediate. After acidic workup, 3-substituted indoles were isolated. For substrates bearing simple alkyl groups, a substantial amount of tetrahydroquinoline derivatives were generated through 6-endo-trig cyclization. This undesired cyclization was suppressed by using an excess amount (five equivalents based on o-alkenylphenyl isocyanide) of ethanethiol instead of tri-n-butyltin hydride. The 2-stannylindole intermediates proved to be a suitable substrate for Stille coupling, giving 2,3 disubstituted indoles in a one-pot procedure. In addition, the 2-stannylindole intermediates could be converted to 2-iodoindoles by treatment with iodine or N iodosuccinimide. The 2-iodoindoles thus obtained served as good substrates for Heck reactions, Stille couplings, Suzuki couplings, and palladium-mediated carbonylations, to afford a variety of 2,3-disubstituted indoles. The utility of this protocol was demonstrated by application to synthetic studies on gelsemine and discorhabdin A, and the total synthesis of an aspidosperma alkaloid, (-) vindoline. PMID- 11933259 TI - Preorganization and protein dynamics in enzyme catalysis. AB - Recently, an alternative has been offered to the concept of transition state (TS) stabilization as an explanation for rate enhancements in enzyme-catalyzed reactions. Instead, most of the rate increase has been ascribed to preorganization of the enzyme active site to bind substrates in a geometry close to that of the TS, which then transit the activation barrier impelled by motions along the reaction coordinate. The question as to how an enzyme achieves such preorganization and concomitant TS stabilization as well as potential coupled motions along the reaction coordinate leads directly to the role of protein dynamic motion. Dihydrofolate reductase (DHFR) is a paradigm in which the role of dynamics in catalysis continues to be unraveled by a wealth of kinetic, structural, and computational studies. DHFR has flexible loop regions adjacent to the active site whose motions modulate passage through the kinetically preferred pathway. The participation of residues distant from the DHFR active site in enhancing the rate of hydride transfer, however, is unanticipated and may signify the importance of long range protein motions. The general significance of protein dynamics in understanding other biological processes is briefly discussed. PMID- 11933261 TI - Nonlinear organic reactions to proliferate acidic and basic molecules and their applications. AB - Acid amplifiers derived from a certain class of sulfonates suffer from autocatalytic decomposition in the presence of a strong acid to give corresponding sulfonic acids, which catalyze the decomposition of the parent sulfonates, leading to the liberation of more of the same sulfonic acids in an exponential manner. Five types of acid amplifiers displaying acid proliferation reactions are presented. A certain type of carbamate exhibits autocatalytic fragmentation to give the corresponding aliphatic amine and olefin together with carbon dioxide, whereas the generated amine is able to act as a catalyst for the fragmentation so that the carbamates are referred to as base amplifiers. Applications of acid and base amplifiers to photofunctional materials, including photoresists, are described as a consequence of the combination of the molecular amplifiers with photoacid as well as photobase generators. Practical applications of acid proliferation reactions in polymer films are discussed. PMID- 11933263 TI - Back to the negative Pap test: behind the scenes at Bethesda 2001. PMID- 11933265 TI - Quantitative FISH analysis on interphase nuclei may improve diagnosis of DNA diploid breast cancers. AB - The detection of DNA aneuploid cells using flow cytometry is an indication for the presence of tumor cells, but when DNA diploid cells are found in 25-33% of the cases, the diagnostic and prognostic significance of DNA ploidy is more limited. We analyzed interphase nuclei after in situ hybridization and using image cytometry on 50 breast tumors with diploid DNA content to investigate whether early chromosome rearrangements were detectable and if their occurrence was clinically significant. Imbalances between the two arms of chromosome 1 were found in 55% of the cases and values ranged from 1.5-3.0. Comparison with histological data showed that Grade I tumors mainly have imbalances (67%) and that Grade III tumors were mainly without the imbalance (67%), whereas Grade II tumors were intermediate (50% imbalance). These data suggest that the diagnosis of DNA diploid cases may be improved by using interphase FISH. In addition, the data also indicates that early breast tumors may have different genetic origins, which is important in the comprehension of tumor malignancy in early stages, especially for preinvasive lesions. PMID- 11933266 TI - Significance of the cytologic diagnosis of endocervical glandular involvement in high-grade squamous intraepithelial lesions. AB - The cytologic criteria for the diagnosis of endocervical gland involvement (EGI) by high-grade squamous intraepithelial lesions (HGSILs) have been described, and this diagnosis occasionally is made. This study evaluates the accuracy of a cytologic diagnosis compared with that of follow-up cone biopsies. Twenty-eight patients with Papanicolaou (Pap) smear diagnoses of HGSILs with EGI, with follow up cone biopsies, were identified from New York University computerized files. Results were compared with those of a control group of 28 patients showing cervical intra-epithelial neoplasia grades II/III (CIN-II/III), irrespective of previous Pap smear findings. On subsequent cone biopsy samples, 26 of the 28 study cases showed signs of HGSIL. Of these 26 patients, 17 (65%) showed evidence of HGSIL with EGI. Among the 28 control cases, 20 (71.4%) had EGI on the cone biopsies (P = NS). We also examined previous Pap smear findings in a control group of 42 cone biopsies with CIN-II or CIN-III, with or without EGI. EGI was diagnosed in previous Pap smears in 3 of the 31(10%) cases that showed signs of EGI on cone biopsies and in 2 of the 11 cases (18%) that did not evidence EGI on subsequent cone biopsies (P = NS). In our experience, the cytologic diagnosis of EGI on Pap smears did not identify a group of patients with increased frequency of EGI on subsequent cone biopsies. PMID- 11933264 TI - Cytology of primary central nervous system neoplasms in cerebrospinal fluid specimens. AB - Although two-thirds of tumors occurring in the central nervous system (CNS) are primary neoplasms, only 10% of positive cerebrospinal fluid (CSF) specimens are from primary CNS tumors. In this study, we reviewed the cytologic findings of 21 positive CSF specimens from primary CNS tumors. A computer search identified 21 cases of positive CSF specimens from patients with primary CNS tumors from the archives. Follow-up included review of medical charts and histologic correlation. The specimens were from 20 patients (9 females and 11 males). Their ages ranged from 6-83 yr, old with a mean of 30 yr. The cases included 9 medulloblastomas, 7 gliomas (3 glioblastoma multiformes, 2 anaplastic astrocytomas, and 2 ependymomas), 2 germinomas, 2 non-Hodgkin's large B-cell lymphomas, and 1 ganglioneurocytoma. Two cases were classified as suspicious and the remaining as positive for malignancy. Immunocytochemistry was employed in 3 cases to support the cytologic diagnosis. These cases included one large-cell lymphoma (leukocyte common antigen-positive), one germinoma (placental alkaline phosphatase positive), and the ganglioneurocytoma (neuron-specific enolase- and synaptophysin positive). There were no false-positive cases. Our results suggest that positive CSF cytology in patients with a primary CNS tumor is a reliable indicator of malignancy and reflects leptomeningeal involvement. The use of immunocytochemistry is helpful in confirming the cytologic impression in some cases. PMID- 11933267 TI - Fine-needle aspiration cytology of phyllodes tumors. AB - Breast lesions with a significant spindle cell or mesenchymal component are not commonly encountered in fine-needle aspiration (FNA) cytologic material and include a heterologous variety of benign and malignant conditions, with phyllodes tumors (PTs) being the foremost differential diagnostic consideration. This study comprises 28 tumors diagnosed histologically as PT in which FNAC material was available for review. Histological sections and cytological smears from these cases were retrieved and subjected to detailed morphological review. Cytological parameters assessed included ratio of stroma to epithelium, pattern characteristics and cytological characteristics of the stromal, and epithelial components and the background cells. Large and hypercellular stroma fragments, dissociated spindle and plump stromal cells, often accompanied by large, folded sheets of epithelium were cytological features that characterized PT. Smears from malignant PT showed predominantly or solely mesenchymal components. FNAC was a highly reliable procedure for the diagnosis of PT, giving an accuracy rate of 92.8%. PMID- 11933268 TI - Nested PCR for diagnosis of tuberculous lymphadenitis and PCR-SSCP for identification of rifampicin resistance in fine-needle aspirates. AB - An accurate diagnosis of tuberculosis and multidrug resistance is important for the control of tuberculosis, which remains a major public health problem. Fine needle aspiration (FNA) has provided an alternative tool for bacterial examination. This study was performed to investigate the usefulness of one-step polymerase chain reaction (PCR) and PCR-SSCP as a routine test for the detection of Mycobacterium tuberculosis and rifampicin-resistant strain in FNA. Ziehl Neelsen stain (Z-N) and PCR were processed using the aspirates of tuberculous lymphadenitis for the detection of M. tuberculosis. PCR-SSCP was done for the identification of rpoB mutation. M. tuberculosis was detected in 49/63 (77.8%) by PCR and 25/63 (39.7%) by Z-N. There were 26 cases with PCR(+)/Z-N(-) and two cases with PCR(-)/Z-N(+). Twelve cases showed negativity against both. In 7/22 (31.8%), rpoB mutation was observed. In conclusion, PCR is more sensitive in the detection of M. tuberculosis in FNA than Z-N. PCR-SSCP could also be used in FNA in the prediction of multidrug resistance. PMID- 11933269 TI - Diagnosis of soft tissue tumors by fine-needle aspiration with combined cytopathology and ancillary techniques. AB - The diagnosis of mesenchymal neoplasm by fine-needle aspiration biopsy (FNAB) has presented a diagnostic challenge. Most reports claim an accuracy approaching 95%, but while they distinguish benign and malignant lesions, the most problematic group, the intermediary malignant group, is omitted. The purpose of this study was to determine whether rapid cytologic diagnosis of soft-tissue tumors could guide surgeons in therapeutic decisions without the need for a tissue biopsy. Ninety-four FNA cytologic specimens were examined by the National Soft Tissue Consortium of Hungary and compared with the corresponding histology. Ordinary lipomas were excluded. Morphologic evaluation was supplemented by ancillary techniques such as fluorescence in situ hybridization (FISH), DNA cytometry, and immunocytochemistry. From a practical clinicopathological point of view, the cases were grouped in the following categories: 1) tumors with definitive diagnosis: a) high-grade malignant neoplasms (high-grade sarcomas, metastatic carcinomas, lymphoma), b) tumors with precise histogenetic origin by cytogenetics, c) benign tumors; 2) tumors of questionable nature. In the first group there were 74 tumors: 22 high-grade sarcomas, six metastatic carcinomas, one malignant lymphoma, 16 malignant tumors in which the precise histogenetic origin could be established by cytogenetic studies, and 29 benign soft-tissue tumors other than lipomas. In the second group there were 20 tumors comprising benign and malignant soft tissue tumors of low grade, wherein the precise nature of the neoplasm could not be established with confidence on cytologic study, even using ancillary techniques. FNAB of soft-tissue tumors combined with ancillary techniques should be considered a viable diagnostic technique for therapeutic protocols. Although the second group is fairly large, we have reliable, well characterized categories which provide great freedom for preoperative and surgical treatment, thus providing the best chance for healing. PMID- 11933270 TI - Diagnosis of histoplasmosis in urine cytology: reactive urothelial changes, a diagnostic pitfall. Case report and literature review of urinary tract infections. AB - Histoplasmosis not uncommonly causes systemic infection, particularly in immunocompromised patients. In systemic infection, the urinary tract is often involved, although the diagnosis of histoplasmosis in urine cytologic specimens has never been reported. Urinary tract histoplasmosis may present with gross hematuria, raising clinical suspicion for malignancy. The index case presented with intermittent gross hematuria, suprapubic pain, significant weight loss, hoarse voice, and a painful tongue ulcer. Examination of the patient revealed an ulcerated tongue lesion, an anal ulcer, a polypoid lesion on the vocal cord, and cystoscopic examination of the urinary bladder revealed erythematous patchy areas. Surgical biopsy sections from the vocal cord and tongue lesion were diagnostic of histoplasma infection. Urine cytologic examination showed atypical urothelial cells suspicious for malignancy. However, fungal stains performed on the urine specimen showed histoplasma organisms. We conclude that with a high index of suspicion, and the use of special stains, histoplasma organisms can be identified in urine. PMID- 11933271 TI - Myxopapillary ependymoma: cytomorphologic characteristics and differential diagnosis. AB - Myxopapillary ependymoma (ME) is a rare tumor with a predilection for sacrococcygeal area of adults. We present the cytomorphology of one such case in a 21-yr-old man, diagnosed by fine-needle aspiration (FNA). The tumor disclosed biphasic morphology with nests and aggregates of epithelioid malignant cells as well as branching cords of myxohyaline material. Most noticeable was the presence of distinct hyaline globules surrounded by neoplastic cells. The differential diagnosis of ME includes chordoma, adenoid cystic carcinoma, mucinous adenocarcinoma, and germ cell tumor. An accurate preoperative diagnosis of ME on FNA can be achieved based on its distinctive cytomorphologic features. PMID- 11933272 TI - Fine-needle aspiration biopsy in diagnosis of follicular lymphoma: cytomorphologic and immunohistochemical analysis. AB - Follicular lymphoma is a relatively uncommon type of non-Hodgkin's lymphoma (NHL) thought to derive from follicular center cells. There are limitations to fine needle aspiration in the diagnosis of follicular NHL, and very few studies are available on it. We describe here the cytomorphologic features of eight biopsy proven cases of follicular NHLs and five biopsy-proven cases of intermediate grade NHLs. Five cases of reactive lymphoid hyperplasia (RLH) diagnosed on cytology were also studied cytomorphologically. Image morphometry was done in follicular and intermediate-grade NHLs, immunocytochemistry was done in follicular and intermediate-grade NHLs, and immunocytochemistry for bcl-2 expression was studied in five cases of follicular NHLs. The aspirate smears of follicular NHL showed the presence of higher grades, many to abundant (++ to +++) monomorphic lymphoid aggregates, with an increased number of mast cells. Lymphohistiocytic tangles and tingeable body macrophages were not observed in any of the cases. Cytomorphologically, all the cases of intermediate-grade NHL showed a predominantly monomorphic population of lymphoid cells with absent or few grades (+) of lymphoid aggregates. Image morphometric data analysis of each cell parameter for follicular and intermediate-grade NHLs were found to be statistically insignificant (P > 0.05) and hence of little help in diagnosis of follicular vs. low-grade NHLs. Immunocytochemistry for bcl-2 expression in follicular NHL showed variable results, ranging from 0-90% (expressed as bcl-2 index). Certain characteristic cytomorphological features such as the presence of monomorphic lymphoid cell aggregates and an increased number of mast cells are soft indicators for the diagnosis of follicular NHL on aspiration smears. Immunocytochemistry for bcl-2 expression may be of help in cases with clinical and cytological suspicion of lymphoma. Image morphometry has a small role in this regard. PMID- 11933273 TI - Hepatocellular carcinoma-fibrolamellar variant: cytopathology of an unusual case. AB - The fibrolamellar variant of hepatocellular carcinoma (FL-HCC) is an uncommon visceral malignancy with a distinctly better prognosis. An accurate initial diagnosis, therefore, is essential for prognostic and therapeutic reasons. This case report describes the cytomorphology of an unusual case of FL-HCC in a 13-yr old boy who was found to have a hepatic mass with prominent intraluminal growth pattern seen predominantly in the right hepatic duct. A diagnosis of FL-HCC was originally suspected on bile duct brushings obtained during endoscopic retrograde choleangiopancreaticogram (ERCP) and subsequently confirmed on fine-needle aspiration (FNA) and surgical resection of the hepatic tumor. The cytomorphologic profile of FL-HCC is distinctive in the right clinical setting, allowing an accurate preoperative identification of this malignancy on biliary brushing or FNA. PMID- 11933274 TI - Comparison of in-house polymerase chain reaction method with the Roche Amplicor technique for detection of Mycobacterium tuberculosis in cytological specimens. AB - Two techniques have been approved by the United States FDA for diagnosis of tuberculosis in smear positive sputa: LCX M. tuberculosis, a ligase chain reaction procedure manufactured by Abbott Laboratories, and Amplicor, a polymerase chain reaction (PCR) procedure manufactured by Roche. However, these commercial methods are expensive and beyond the reach of laboratories in most developing countries. We compared the Roche Amplicor kit with an in-house PCR using a primer set for Mycobacterium tuberculosis/bovis directed at MPB 64 protein gene. It was able to distinguish between M. tuberculosis, M. avium, M. gordonii, M. intracellularae, and M. kansasii. Fifty-seven cytological samples were submitted to the laboratory for molecular diagnosis of M. tuberculosis. Both procedures were run on every sample submitted and the two methods agreed completely. The custom-made method is less expensive than the commercial technique. PMID- 11933275 TI - Aspiration cytology of epidermoid cyst of terminal phalanx. PMID- 11933276 TI - Cytology of high-grade squamous intraepithelial lesion in Japanese-Brazilian women with HIV infection with polymerase chain reaction-assisted human papilloma virus detection. PMID- 11933277 TI - Recommendations for the quantification of the survey process. PMID- 11933278 TI - Using complementary and alternative techniques--boosting immunity. PMID- 11933279 TI - Cost cutting: balancing care needs with successful strategies. PMID- 11933280 TI - Randomised controlled trial of two pressure-relieving systems. AB - The primary objective of this randomised controlled trial was to determine whether there were significant differences between two pressure-relieving systems. A secondary aim was to investigate whether the availability of extra pressure-relieving equipment would reduce the incidence of ulcers in an acute hospital setting. A total of 141 patients in a care-of-the-elderly unit, who were assessed to have a high risk of developing pressure ulcers using the Waterlow score, were recruited; 70 were nursed using Huntleigh Nimbus 3 in conjunction with the Aura cushion (Group A), and 71 using the Pegasus Cairwave Therapy System in conjunction with the Proactive 2 Seating cushion (Group B). The main outcome measure was visual assessment, supported by a photographic record. There were three main findings: for non-heel ulcers and overall improvement, there was no statistically significant difference between the two products tested; for heel ulcers there was a significant difference (P = 0.019) with more patients healing in Group A than in Group B. The average length of stay of patients who completed the trial was 21.6 days (Group A) and 21.7 days (Group B) for patients completing a live (range 1-121 days) and for patients who died 29.7 days (Group A) and 24.3 days (Group B). Routine monitoring showed that, before the trial, the incidence of hospital-acquired pressure ulcers (Torrance grade 2+) was 0.2%; during the trial, this dropped to 0.13%. The study showed differences in the efficacy of different mattress products; with a sufficiently large study, it is possible to demonstrate statistically significant results. Provision of extra pressure relieving equipment can reduce the incidence of pressure ulcers but may not influence length of stay. PMID- 11933281 TI - Alginate dressings in surgery and wound management--Part 1. AB - Large quantities of alginate dressings are used each year to treat exuding wounds, such as leg ulcers, pressure sores and infected surgical wounds. Originally these dressings were a loose fleece formed primarily from fibres of calcium alginate. More recently they have been developed so that the fibres have been entangled to form a product with more cohesive structure, which increases the fabric's strength when it is soaked with exudate or blood. Some products also contain a significant proportion of sodium alginate to improve the gelling properties of the dressing in use. Other dressings have been produced from freeze dried alginate. Once in contact with an exuding wound, an ion-exchange reaction takes place between the calcium ions in the dressing and sodium ions in serum or wound fluid. When a significant proportion of the calcium ions on the fibre have been replaced by sodium, the fibre swells and partially dissolves forming a gel like mass. The degree of swelling is determined principally by the chemical composition of the alginate, which depends on its botanical source. Although it is recognised that the differences between the various brands of dressings may influence their handling characteristics--particularly when wet--it is generally assumed that these differences are of limited relevance to the dressing's performance clinically or at a cellular level. There is some evidence to suggest, however, that these assumptions may be wrong and that alginates may influence wound healing in a number of ways not yet fully understood. This three-part review of the literature encompasses the history, origin, structure, chemistry and clinical applications of alginates and alginate dressings. This review reveals that, despite their widespread use, alginates have been the subject of very few well-controlled clinical studies. There is fairly convincing evidence, however, that they do offer advantages over more traditional dressings for at least some clinical indications. It has also become obvious that there is a general lack of understanding about the importance of secondary dressing systems that must be used in with alginate dressings. Careful examination of the design and outcomes of the published studies suggests that the choice of both the primary alginate dressing and the secondary dressing can play a major role in determining treatment outcomes. PMID- 11933282 TI - Implementation of patient-held records in diabetic foot care. PMID- 11933283 TI - The prevention of pressure ulcers through risk assessment. PMID- 11933284 TI - Selecting the most appropriate armchair for patients. PMID- 11933285 TI - The use of a non-adherent silicone dressing in arterial leg ulceration. PMID- 11933286 TI - Managing pressure ulcers in a nursing home patient. PMID- 11933287 TI - A simple severity index for pressure ulcers. AB - The aim of this study was to design and test a pressure ulcer severity score based on assessment by experienced clinicians. Fifty pressure ulcers were each evaluated independently by seven experienced clinicians, using a scale that ranged from 0 (mildest damage) to 10 (most severe). Ulcer characteristics were analysed to determine which factors related to the assessments. A second set of 50 ulcers was used to test the validity of the resulting scoring system. Inter- and intra-observer reliability were tested in a further 10 pressure ulcers. A multivariate linear model was produced, based on the following three variables: colour (CO-matched-matched) in the wound base; depth (DE) of the ulcer; and mean diameter (D) of the wound ('CODED'). The coefficients for each variable were rounded to yield points in an additive equation, the CODED score. This score had a high correlation (r = 0.92, P < 0.001) with the second set of evaluations, used to test the score performance. The CODED score is a simple and valid tool to quantify the progress of a pressure ulcer. It has a high correlation with the severity assessment of experienced clinicians and is reliable when used by non experienced observers. PMID- 11933288 TI - Acute and chronic wounds: differences in self-reported health-related quality of life. AB - Seventeen patients (10 female) with acute wounds (pilonidal sinus) and 32 patients (19 female) with chronic wounds (venous leg ulcers) attending a specialist wound clinic completed a condition-specific health-related quality of life (HRQoL) tool on two occasions five to seven days apart. The ratings from the two groups were statistically different for two sub-scales, with those with acute wounds rating themselves more positively. There were no differences between the groups for overall HRQoL or satisfaction with HRQoL--although there was a trend for those with chronic wounds to rate their overall HRQoL as higher than those with acute wounds, possibly indicating adaptation to their health status. While possible demographic and aetiological differences must be borne in mind, the tool used in this study appears to be sensitive to different types of wound. PMID- 11933289 TI - Cigarette smoking and its effects on wound healing. PMID- 11933290 TI - Wound care guidelines and formulary for community nurses. AB - Community nursing is experiencing significant change as a result of developments such as improved technology, care in the community and earlier discharge of patients from hospital. Because of this, increasingly complex clinical care is required in the community, and it has been noted that community nurses are 'under considerable pressure' and show 'evidence of high stress and low morale'. Wound care is one area in which community nurses constantly battle to keep abreast of continual change. Growing product availability and diversity of use, changes in dressing techniques and the ever-increasing costs associated with wound care mean decision-making in wound care is often a complex task. In the Grampian region, a handbook of evidence-based practice guidelines with a product formulary was developed and distributed to all community nurses. The handbook was designed to ease the decision-making process by evaluating evidence-based practice and local preferences to recommend and guide nurses towards effective clinical practice and cost efficiency. All grades of district nurse in the region have been issued with their own copy of the handbook. It is presented in an A5 ring-binder format to make it easy to carry and to facilitate updating using loose-leaf inserts. The use of logos, extra information boxes and colour coding makes it easy for users to find specific areas of interest in the handbook. The success of the handbook has led to debate on the potential for development of a similar resource for use by practice nurses and in local community hospitals. PMID- 11933291 TI - Pulse oximetry: a new tool to assess patients with leg ulcers. AB - The objective of the study was to investigate pulse oximetry as a guide to assessing patients with leg ulcers before treatment. Graduated elastic compression is the treatment of choice for uncomplicated venous leg ulcers, but is contra-indicated in patients with significant arterial disease. The standard assessment of arterial insufficiency by Doppler ultrasound ankle branchial pressure index (ABPI) has shortcomings which prompted this investigation of pulse oximetry as a possible additional, or alternative, method of assessment of patients with leg ulcers, prior to treatment with compression. The study, carried out on a population of patients attending hospital leg ulcer clinics, was designed to evaluate pulse oximetry assessment in the selection and monitoring of patients with venous leg ulceration leading to a prospective controlled study of ulcer healing in groups of patients with reduced and normal ABPI, selected for compression therapy by pulse oximetry criteria. Outcome measurement required follow-up of patients selected for compression therapy by pulse oximetry to record time to healing and rate of healing of leg ulcers. Results from the study show a fair correlation between the toe-finger oximetry index (TFOI) and Doppler ABPI. There is no difference between ulcer healing in patients with reduced and normal ABPI selected for treatment on the basis of pulse oximetry maximum compression pressure (MCP). In conclusion, pulse oximetry is an aid to the selection of patients who will benefit from compression therapy, but would be excluded on the basis of Doppler ABPI. PMID- 11933293 TI - The Dutch experience of pressure ulcers--a personal view. PMID- 11933292 TI - Alginate dressings in surgery and wound management: Part 2. PMID- 11933294 TI - Hypergranulation tissue at gastrostomy sites. PMID- 11933295 TI - Chronic leg ulcers in Sweden: a survey of wound management. AB - The aim of this study was to survey management of leg ulcers by staff in Swedish primary healthcare (PHC) and home care services (HCS) run by the municipalities (n = 933), with emphasis on wound care and education. A questionnaire was completed by 933 staff (78% response rate). Diagnostic investigation and documentation in specific wound records was performed by 46.6% of respondents in PHC and 12.8% in HCS, most commonly by nurses in PHC compared to nurse auxiliaries (NAs) in the HCS. Topical treatment was most commonly chosen by nurses (82.1%). Nurse auxiliaries in PHC had greatest access to structured wound management programmes (40.0% compared with 30.1% in HCS, p < 0.05). Instruction courses, mainly organised by pharmaceutical companies (43.2%), were the most common form of education (20.4%) identified. The most time-consuming wound dressings and the highest number of patients treated at home and by NAs were found in the municipalities. For topical treatment 29 products were identified. Nurses in both PHC and the HCS used a wider range of products, e.g. hydrocolloids and absorptive dressings, than NAs in nursing homes, who used dry gauze/protective dressings and wet saline gauze to a greater extent. Substantial differences in qualifications, wound management experience and resource utilisation were found between staff in PHC and the HCS. PMID- 11933296 TI - Studies of a new multi-layer compression bandage for the treatment of venous ulceration. AB - This study aimed to develop an alternative graduated compression bandage for the treatment of venous leg ulcers. Alternative bandage components were identified and assessed for optimal performance as a graduated multi-layer compression bandage. Subsequently the physical characteristics and clinical efficacy of the optimal bandage combination was prospectively examined. Ten healthy limbs were used to develop the optimal combination and 20 limbs with venous ulceration to compare the physical properties of the two bandage types. Subsequently 42 consecutive ulcerated limbs were prospectively treated to examine the efficacy of the new bandage combination. The new combination produced graduated median (range) sub-bandage pressures (mmHg) as follows: ankle 59 (42-100), calf 36 (27 67) and knee 35 (16-67). Over a seven-day period this combination maintained a comparable level of compression with the Charing Cross system, and achieved an overall healing rate at one year of 88%. The described combination should be brought to the attention of healthcare professionals treating venous ulcers as a possible alternative to other forms of multi-layer graduated compression bandages pending prospective, randomised clinical trials. PMID- 11933297 TI - The use of pressure therapy to treat hypertrophic scarring. PMID- 11933298 TI - Infection control: everyone's responsibility. PMID- 11933299 TI - Alginate dressings in surgery and wound management: Part 3. PMID- 11933300 TI - New hydrofibre and hydrocolloid dressings for chronic wounds. PMID- 11933301 TI - High-pressure water jet injuries: a surgical emergency. AB - Injuries from high-pressure jet devices are surgical emergencies characterised by small entry wounds with extensive internal damage. Three factors are involved in these injuries. Physical injury can include local soft tissue disruption and vascular and nerve damage; chemical properties of the injectate can exacerbate compressive vascular injuries with increased oedema and inflammation; water-jet injuries can be contaminated by virulent organisms and foreign matter which can lead to unusual infections. Management is usually similar to that for injuries caused by high-velocity missiles, and involves aggressive debridement, irrigation and decompression followed by careful monitoring, and appropriate antibiotic therapy. PMID- 11933302 TI - Allergic reactions to hydrogels. PMID- 11933304 TI - Evidence and wound care: what is it? PMID- 11933303 TI - A clinical evaluation of the Nimbus 3 alternating pressure mattress replacement system. AB - This study assessed the clinical effectiveness of the Nimbus 3 alternating pressure mattress replacement system (APMRS) on pressure ulcer healing and comfort in subjects > or = 65 years, with at least a Grade 2 ulcer and some mobility problems. Twelve patients in a hospital setting were randomly allocated to the Nimbus 3 or another APMRS, and 20 residents in a nursing home setting to the Nimbus 3 or an alternating pressure mattress overlay. Wound surface area (WSA) (cm2) was recorded twice weekly and comfort once weekly. In the hospital setting, there were no significant differences between groups in the reduction in WSA per day. In the nursing home setting, though subjects on Nimbus 3 had significantly more pressure ulcers at baseline, there were no significant differences between groups in the reduction in WSA per day. Nimbus 3 was statistically more comfortable than control surfaces. The study's sample size has not shown the products were different with regard to clinical effectiveness. However, it might serve as a pilot for a larger, multi-centre RCT aimed at establishing the efficacy of a pressure-relieving (PR) device on pressure ulcer healing. PMID- 11933305 TI - Treatment of diabetic foot ulcers with hyperbaric oxygen. AB - The delivery of oxygen to the wound site is crucial in healing diabetic foot ulcers, and impairment of this process in people with diabetes leads to delayed wound repair. Hyperbaric oxygen therapy works by elevating the plasma oxygen level. Fibroblasts synthesise and modify collagen, and both these activities require relatively high partial pressures of oxygen. Hyperbaric oxygen can promote healing by stimulating fibroblast activity and collagen formation. Increasing oxygen tensions also has a direct and toxic effect on anaerobes, therefore hyperbaric oxygen therapy has a special role in treating diabetic foot infections. Studies on hyperbaric oxygen therapy in general show it to be a beneficial adjunctive therapy for diabetic foot ulcers. However much of the work is anecdotal and more controlled trials are required. PMID- 11933306 TI - The effect of a radiant heat dressing on pressure ulcers. AB - The use of heat in wound healing has been demonstrated to aid oxygen flow and hence healing in acute wounds. However, the situation in chronic wounds is less clear. This study was designed to investigate the benefits of using a radiant heat therapy system in the treatment of Stage 3 and 4 pressure ulcers. Despite randomisation, patients receiving radiant heat therapy were more infirm than those receiving standard treatment. This prospective, single-centre, randomised trial resulted in an accelerated rate of healing for those receiving heat therapy compared to a standard treatment: time difference to 75% of original area = 6.4 days (p = 0.057), to 50% of original area = 9.6 days (p = 0.039), time to 25% = 7.2 days (p = 0.01). This new development warrants further investigation to fully assess the role of a thermoregulation system in chronic wound healing. PMID- 11933307 TI - Blood flow monitoring by pulse oximetry during a forearm flap elevation. AB - The Allen's test for blood flow is performed before a forearm flap is elevated to examine vascularization through the ulnar artery. In this study, to ensure the survival of the donor site when a forearm flap was elevated, we monitored patients using a pulse oximeter. From oral cancer patients who were undergoing reconstructive surgery using the forearm flap, twenty patients were randomly selected to be served as subjects. A pulse oximeter probe was attached to the thumb on the side from which a skin flap was to be elevated. Oxygen saturation was measured when the tourniquet was released, and the radial artery was clamped and severed. The results showed that oxygen saturation temporarily decreased when the radial artery was ligated, but it recovered in one to two minutes. PMID- 11933308 TI - Gender differences in oral health behavior and general health habits in an adult population. AB - This study aimed to evaluate gender differences in oral health behavior and general health habits in adults. The subjects were 207 males and 196 females aged 20-64 yrs who were public officials in the city or town administrations in Chiba Prefecture, Japan. The questionnaire survey included three items: (1) self assessment of oral health status, (2) oral health behavior and (3) general health habits. Statistical analysis was performed using the chi-square test for differences of responses between males and females. The proportion of subjects with cognition of symptoms of oral disease ranged from 14.3 to 23.0%. The percentage of those who had not visited a dentist in the last year were 52.7% for males and 36.7% for females (p < 0.01). Subjects who brushed their teeth almost every day at bed time were 60.9% of males and 88.8% of females (p < 0.01). A comparison of the numbers of positive responses regarding general health habits found no differences in the distribution of general health habits score between males and females. Examining the relationship between oral health behavior and general health habits revealed that males with general habit high scores tended to have positive oral hygiene behavior. These results support the thesis that gender specificities in oral health depend on individual attitudes to oral health and dental utilization. In addition, understanding the cognitive factors of males and females would accelerate dental approaches to modifying oral health behavior of both groups, thus contributing to lifelong health maintenance. PMID- 11933309 TI - An electromyogram study on mandibular movement in unilateral cleft lip and palate patients before and after orthodontic treatment. AB - The purpose of this study is to investigate masticatory muscle function in subjects with unilateral cleft lip and palate compared with normal occlusion and the extents of improvement before and after orthodontic treatment. Subjects were twenty pretreatment patients, thirty posttreatment patients and ten controls. Electromyograms were recorded from their masticatory muscles during masticatory movement and tapping movement. Reduction of duration/stroke ratio (D/S) was observed in electromyograms obtained during masticatory movement after orthodontic treatment. The reduction was especially notable in the masseter muscle. In spite of a significant reduction in coefficient of variation (CV) values, a significant difference between the posttreatment group and the control group indicated some persisting irregularity in masticatory movement. Electromyograms taken during tapping movement showed no change in latency in the posttreatment group, but duration of the silent period (SP) was shortened and SP appearance increased. The findings outlined above reveal electromyographically clear improvements in masticatory muscle functions and jaw reflex mechanisms after orthodontic treatment. Nevertheless, parameters for subjects with cleft lip and palate still differed from those for controls with normal occlusion. The influence of plastic surgery in subjects with the defects discussed here causes maxillary retrusion, which in turn results in skeletal malocclusion. Orthodontic treatment should be designed to compensate this dentally and alveolarly. This design and the need to improve masticatory functions would contribute to eliminate the extreme difficulty of the therapeutic process. PMID- 11933310 TI - The horizontally impacted maxillary canine situated in a labial position. AB - Surgical management of unerupted teeth depends upon a thorough understanding of anatomic, physiologic and pathologic factors. Attention has been given to problems of eruption in the maxillary anterior region. It is a region where a variety of anomalies occur. Since the maxillary anterior region influences appearance so greatly, early detection of difficulties and careful planning and treatment can be extremely beneficial to patients. The purpose of this case report is to present a case of maxillary permanent canine impaction in a horizontal displacement that developed after loss of the deciduous canine to chronic apical periodontitis, and incomplete root resorption of the deciduous canine. PMID- 11933311 TI - Metachronous multiple carcinoma of lip after surgery for gingival carcinoma: a case report. AB - A case of multiple carcinoma of the lower lip probably induced by repeated bite trauma due to lack of sensation is reported. A reduction in immunocompetence by carboplatin (CBDCA) might be an additional condition for the carcinogenesis. This case suggests that the reconstruction of the sensory nerve is important after resection of a malignant tumor of the head and neck region. PMID- 11933312 TI - Sildenafil induced priapism. AB - A 53 year-old Japanese man was referred to our hospital for persistent priapism, which had been induced by 200 mg (usual dose 25-50 mg) of sildenafil citrate (Viagra) three days earlier. He had a history of erectile dysfunction and had undergone penile injection therapy elsewhere; however, he had not used injection therapy this time. He obtained sildenafil personally without a doctor's prescription. He had not taken any other drugs that affect the metabolism of sildenafil, nor did he have any medical complications that might induce priapism. Since needle aspiration and irrigation were ineffective as first line therapy, surgical treatment was indicated to relieve the condition; a incision of tunica albuginea of both corpora cavernosa was made, and vigorous irrigation of saline washed out the blood clots. This is the first case report of priapism induced by sildenafil. Although this drug can be obtained through private commerce, it should be used under professional guidance as its abuse may lead to severe morbidity. PMID- 11933313 TI - Signs and symptoms in ophthalmology. PMID- 11933314 TI - Relationship between eye condition and ADHD. PMID- 11933315 TI - Consumers are being inundated with advertisements touting the benefits of LASIK eye surgery. PMID- 11933316 TI - Getting the clear picture on LASIK eye surgery. PMID- 11933317 TI - Medical billing outsourcing: is it worth the risk? PMID- 11933318 TI - Device allows adjustments for lumboperitoneal shunting. PMID- 11933319 TI - Identifying and treating ocular emergencies. AB - The nurse's role in addressing ophthalmic emergencies can determine the urgency of the treatment and, in some cases, the final visual outcome. It is important to understand the spectrum of traumatic eye injuries so the patient will be better cared for and the appropriate treatment regimen will be started before permanent vision loss occurs. Assessing visual acuity is key; it must be documented, and if visual acuity cannot be obtained immediately, this fact also must be recorded. PMID- 11933320 TI - Avulsion of the optic disc after a blow to swimming goggles. PMID- 11933321 TI - Advanced manual lensometry: a self-learning guide for evaluating multifocal and specialty lenses, progressive lenses, prisms, and rigid contact lenses. AB - Advanced manual lensometry techniques are required when automated lensometry may not be appropriate for reading special lenses or rigid contact lenses. These advanced techniques are challenging but provide accurate diagnostic and lens verification data. One should never assume that the glasses the patient is wearing match what the doctor prescribed. The prescription should be checked at least once, even when there are no patient complaints. If possible, learn and practice these special procedures under the supervision of an optician or ophthalmic clinical trainer. This will help to verify that you are performing these special techniques correctly. PMID- 11933322 TI - New surgical treatments for macular degeneration. AB - The new surgical techniques described in this article expand the list of new treatments becoming available for people with macular degeneration. PMID- 11933324 TI - Understanding aspheric contact lenses. PMID- 11933323 TI - The nursing process in photodynamic therapy. AB - The results from the TAP Study have been found to be significant, but remember this is not a cure. There are no quick and easy solutions to this devastating disease, which can creep up so suddenly and without warning. Ophthalmologists may be able to stabilize the vision without allowing it to become worse, but the key words are "without allowing it to become worse." Unfortunately, to most patients, it is already worse. Nurses must address the patients' feelings and concerns first, before offering a physical solution that is only a temporary "band aid." After 10 years of working in the area of ophthalmology, specifically low vision, it has come to the authors' attention that a large percentage of patients suffering from AMD display depressive symptoms such as feelings of sadness, worthlessness, and hopelessness, and low self-esteem. Many appear to go through a grieving process. Keeping this is mind, the overall goal in nursing care should be to help patients with AMD use available resources so they have the potential to maintain a satisfying quality of life, physically and emotionally. PMID- 11933325 TI - [Preparation of a research paper for publication: discussion and conclusion]. PMID- 11933326 TI - [Sample taking through central venous catheter for the control of partial thromboplastin time in patients with heparin sodium perfusion]. AB - The need to systematically monitor activated partial thromboplastin time (APTT) in patients undergoing continuous perfusion of heparin sodium (non-fractionated) in order to maintain therapeutic levels of anticoagulation leads to two questions: 1. can blood be withdrawn from the catheter through which the heparin solution is being perfused without altering APTT values? and 2. how much blood should be discarded so that APTT values remain unchanged? To obtain APTT values in these patients, two samples were extracted simultaneously: one through the central venous catheter through which the heparin was being perfused, after previously discarding 10 or 20 ml of blood according to the group to which the patient was assigned and the other through direct venous puncture or through the peripheral catheter inserted in the arm not being used for heparin perfusion. The values obtained by both methods were analyzed for statistical significance. Comparison of the results of APPT in the samples, having previously extracted 10 or 20 ml of blood, with those obtained through venous puncture or through peripheral venous catheter revealed statistically significant differences. These differences were lower when 20 ml were discarded than when 10 ml were discarded. In conclusion, APPT monitoring should be performed whether the sample is obtained through direct venous puncture or through a peripheral catheter inserted in the arm not being used for heparin perfusion. PMID- 11933327 TI - [Work environment at a university hospital: perception of nurses]. AB - Knowledge of the work climate is important in all organizations since the this climate can either contribute to or limit the development and sense of fulfillment of those working within the workplace. The aim of this study was to describe the nurses' perception of the working environment in a university hospital. Two-hundred and thirteen nurses completed a self-administered questionnaire which included the Work Environment Scale (WES) translated to Spanish. The nurses perceived that involvement and cohesion among colleagues was high. The support of the supervisor as well as the autonomy given to nurses in their work was considered moderate. The nurses also highlighted organizational aspects, i.e. planning and efficiency at work and the clarity of what was expected of them in their daily activities. The perception of the work environment differed among the various nursing units. In general, nurses with the fewest years of professional experience perceived the work environment more positively. This study has enabled the determination of a work environment profile of a group of nurses in a university hospital. Moreover, guidance is offered to nurse managers of the nursing units on the positive aspects of the work environment which should be reinforced and on the negative aspects which should be modified in order to create an optimal work environment. PMID- 11933328 TI - [Description of the "new" intensive care unit of the Valladolid Rio Hortega Hospital]. AB - The unit described herein forms part of the Rio Hortega Hospital in Valladolid (Spain) and is situated on the third floor. The building houses several medical and surgical specialities and has been renovated several times; the latest renovations have involved technical and structural renovation of the Intensive Care Unit. The new intensive care unit was inaugurated in February 1998 due to new technical and medical requirements. The aim of improving medical care and the degree of satisfaction among patients and their families has been achieved and the work environment is more congenial and peaceful. The unit is composed of two areas: the multipurpose intensive care unit and the cardiology intensive care unit, edowed with the same monitoring system and apparatus specific to each. Moreover, the unit is composed of an administration area, storerooms and an area for common services (a room for the cleaning of material, waste disposal, lavatories, etc.). The medical team is composed of (a) medical staff-seven staff doctors and five residents; (b) nursing staff: 18 nurses and 13 nurse's aides in the multipurpose Intensive Care Unit; 13 nurses and 8 nurse's aides in the cardiology intensive care unit: one supervisor common to both units; administrative staff: one secretary; one security guard and one cleaner. The distribution of nursing work depends on workload reflected in the patient's daily chart, protocols, nursing care plans and follow-up sheets for the nursing team. Visiting hours are divided between the morning and afternoon and information is given to the patient's relatives by the doctor before the visit. The hospital has three catholic priests who periodically visit the patients. We also try to help when the patients profess other religious faith. The type of patients who are cared for in the unit as well as their distribution according to condition, severity, death, etc, is shown in figures: the data correspond to the period from January 1 1998 to August 1 1999. PMID- 11933330 TI - In-situ irradiation of keloid scars with Nd:YAG laser. AB - In-situ coagulation of keloid with Nd:YAG laser irradiation was carried out in in a cohort study of 17 patients, each with one keloid scar situated on either the sternum (six), abdomen (seven), shoulder (two), hip (one), or ear lobe (one). The duration of scarring was three to 17 years and size varied from 3-8 cm long. One to two doses of laser irradiation was required for complete coagulation. At three months, 10 (58.8%) keloids had completely healed; but in seven (41.1%), 25-50% of residual keloid persisted. Intralesional triamcinolone injection, once in four patients and twice in three patients, produced complete resolution in all seven patients. At 18 months to five years follow-up, 14 patients remain keloid-free but in three keloid recurred and was re-treated with laser coagulation with complete resolution. The treatment was carried out as an outpatient procedure under local anaesthesia. There were no complications. The results of this initial study suggest that Nd:YAG laser irradiation coagulation is effective treatment for keloid scarring. PMID- 11933331 TI - Pinch graft skin transplantation for leg ulcers in primary care. AB - Pinch grafting offers an alternative approach to conventional management of chronic leg ulcers, and is particularly well suited for venous and diabetic ulcers. Relevant literature from 1872-1998 was reviewed. The method of pinch grafting is described here in detail, and is illustrated by two case reports. Since pinch grafting hastens the healing of granulating wounds, is easy to perform and does not require strict immobilisation, a revival of the method, with adaptation for primary care, should be encouraged. PMID- 11933332 TI - Varidase: the science behind the medicament. AB - Varidase is used throughout the world for the topical treatment of purulent and suppurating wounds. Its efficacy is centred on two enzymes, streptokinase and streptodornase. However, these represent only a small proportion of the bulked solid. This article gives an overview of the preparation and mode of action of Varidase, as well as showing some of the research and development that has gone into improving the assessment of its quality and composition. PMID- 11933333 TI - Vacuum-assisted closure to treat deep sternal wound infection following cardiac surgery. AB - Deep sternal wound infection is a devastating and potentially lethal complication following cardiac surgery. Established treatment may involve debridement, packing, delayed closure, plastic reconstruction, re-wiring and irrigation, depending on the severity of infection. The authors prefer early debridement and rewiring with irrigation if possible. If this method fails the results are often poor. Vacuum-assisted closure (VAC) of complex infected wounds has recently gained popularity among various surgical specialties. This article describes the successful application of this novel technique in a patient with infected sternal dehiscence who had failed to respond to conventional treatment. PMID- 11933334 TI - Who sets the research agenda? PMID- 11933335 TI - The effects of radiotherapy on maxillofacial tissue. AB - Radiation therapy for the treatment of maxillofacial tumours can lead to delayed complications. One of the most severe complications is osteoradionecrosis (ORN), which can cause orocutaneous fistulae, exposed mandible and considerable pain. The condition can occur years after radiotherapy and requires aggressive prevention and treatment. Hyperbaric oxygen therapy has been found beneficial for patients with maxillofacial ORN, leading to pain relief, fistula closure and bone coverage. PMID- 11933336 TI - A clinico-epidemiological profile of non-healing wounds in an Indian hospital. AB - One hundred patients with non-healing wounds of more than six weeks' duration were studied for clinico-epidemiological factors and their correlation with healing time. The majority of subjects were aged 30-59 years; those aged 50 and over had significantly delayed healing. Most of the subjects were farmers and physical labourers. Leprosy was the most common cause of non-healing wounds, followed by diabetes, trauma and venous ulcers; the lower limbs were the most common site of involvement. Diabetic wounds were more common in those aged 40 years and above. Diabetic wounds were slower to heal than others. The time to healing was calculated from the day subjects received treatment advice at the wound clinic. Seventy-five of the study group were included for calculation of healing time. The others were lost to follow-up or excluded for other reasons. PMID- 11933338 TI - Why don't health professionals write case studies any more? PMID- 11933337 TI - Developing a new wound care formulary and guidelines. AB - Following an audit of the existing wound care formulary and guidelines, an updated version was developed, containing additional products and information. This was revised following advice from a range of professionals, then pilot tested on ward-based nurses. The nurses found the updated formulary useful, and made recommendations for changes to the wound care ordering system in the trust. Future guidelines will be regularly updated as developments in wound care occur. PMID- 11933339 TI - How effective are prevention strategies in reducing the prevalence of pressure ulcers? AB - The prevalence of pressure ulcers has remained constant at about 7% over the past 20 years, even though considerable time and money has been invested in various prevention strategies. This literature review explores whether pressure prevention programmes can reduce the prevalence rate still lower or whether they are working but are limited by an increasingly aged population and rising patient acuity. PMID- 11933340 TI - Comparison of the healing rates and complications of three four-layer bandage regimens. AB - This randomised controlled study compares the healing rates, complications and patient and staff acceptability of three four-layer bandage regimens for leg ulcers. A total of 149 patients were recruited into the study, of whom 50 received the original Charing Cross system (CX4L), 50 a modified Charing Cross system (Parema) and 49 a commercially available kit, Robinson Ultra Four (Robinson). No significant difference was found in the healing rates of the three systems. Overall 12 weeks' healing was 65%, while the 20-week healing rates for the individual systems were 87% (CX4L), 84% (Parema) and 83% (Robinson). Analysis of known risk factors for delayed healing showed that no bandaging system had an advantage over the others. Staff familiarity resulted in an initial preference for the CX4L but there was no bandage preference by the end of the study. The data suggest that none of the systems has an advantage over the others and that cost savings can be made by pursuing a competitive pricing policy. PMID- 11933341 TI - Pressure ulcer classification: the Torrance system. AB - A wide variety of pressure ulcer classification systems are in use but most have grading systems which are subjective and open to different interpretations. This review discusses the advantages and disadvantages of the Torrance classification system, and argues that this highlights the need for a national grading tool. PMID- 11933342 TI - Fads and fancies. PMID- 11933343 TI - The diagnosis and treatment of pyoderma gangraenosum. AB - Pyoderma gangraenosum is a rare condition and its symptoms are often misdiagnosed. This article presents of an overview of this form of ulceration, and includes a case study showing how it was diagnosed and treated in a patient following an arthroscopy to the left ankle. PMID- 11933344 TI - Honey and the management of infected wounds. PMID- 11933345 TI - The assessment and management of leg ulcers in rheumatoid arthritis. AB - Patients with rheumatoid arthritis appear to be at increased risk of developing chronic leg ulcers. This review identifies the factors that predispose these patients to leg ulceration and highlights how this problem can be managed and the risk of recurrence reduced. PMID- 11933346 TI - The physiology of wound healing. PMID- 11933347 TI - The for-profit providers. PMID- 11933348 TI - Advanced practice nurses in ESRD: varied roles and a cost analysis. AB - The predicted shortage of nephrologists to care for end-stage renal disease (ESRD) patients has prompted examination of the potential roles for physician extenders, such as physician assistants and nurse practitioners or advanced practice nurses (APNs) in the care of these patients. Although the case management model is ideally suited for the care of ESRD patients, little is known about incorporating APNs within this model in either dialysis or transplant programs. Furthermore, the cost-effectiveness of APNs working in dialysis units has been unexplored. We report our experience with APNs working in our dialysis and transplant programs, providing a description of their varied roles in the outpatient and inpatient settings that emphasizes the flexibility of the case management model in the care of ESRD patients. A time log survey completed by dialysis APNs outlines a mode for job responsibilities and shows that 93% of their time is spent in direct patient care. The dialysis cost analysis reveals that employment of a nurse practitioner is cost effective if the dialysis program includes at least 85 chronic dialysis patients. Additional studies of the impact of APNs in ESRD programs is needed. PMID- 11933349 TI - The nephrologist at the cross-roads: physician and manager. PMID- 11933350 TI - Hematocrits show notable improvement for hemodialysis patients. As in previous years, quality of care varies by location. PMID- 11933351 TI - The nephrology social worker as clinician. PMID- 11933353 TI - Improving physician-patient communication can increase success of rehabilitation. PMID- 11933352 TI - Understanding the psyche of the disruptive patient in the dialysis facility. PMID- 11933354 TI - The community and orthodontic care. Part I: community-perceived need and demand for orthodontic treatment. AB - A professionally managed telephone survey was undertaken to assess community perceived need and demand for orthodontic treatment, and to determine the proportion of the community with a history of having some form of orthodontic treatment. The sample included 505 respondents, aged eighteen and over, from metropolitan and non-metropolitan households across the state of Victoria in Australia. The sample distribution had a ninety-five per cent confidence limit with a five per cent margin of error, and closely matched the known population distributions for age, sex and geographical location. From the survey it can be concluded that apparently forty-four per cent of Victorian families include someone who has already received some form of orthodontic treatment. Twenty-five per cent of the survey respondents perceived some need for the treatment of a family member; only fifteen per cent of respondents, however, reported that someone in their family actually wanted treatment. This survey has established baseline values for community perceived need and demand for orthodontic treatment. Use of these values should assist in future resource management within both the public and private sectors. PMID- 11933355 TI - Variations in treatment need using four screening methods. AB - The study models of 100 Grade Seven students were used to compare the Dental Aesthetic Index (DAI), the Dental Health Component (DHC) and the Aesthetic Component (AC) of the Index of Orthodontic Treatment Need (IOTN), and the Danish Ministry of Health (DMH) screening system. The basis for comparison was the agreed subjective assessment of two orthodontists for each subject. Disagreements between the subjective assessment and each screening method were further analysed in an attempt to identify the specific occlusal traits responsible for the disagreement. The DAI under-estimated treatment need in cases with displaced canine teeth, incisor crowding or rotations and increased overbite. The DAI over estimated treatment need in cases with increased overjet in otherwise well aligned arches. The DMH guidelines over-estimated treatment need in cases with increased overjet and crowded arches. The DHC was found to be over-sensitive in cases with increased overjet and contact point displacements greater than 2 mm. The AC under-estimated treatment need in cases with excessive overjet and buccally displaced canines, and over-estimated treatment need in cases with spaced arches and deep overbite. PMID- 11933356 TI - EICO-1: an orthodontist-maintained expert system in clinical orthodontics. AB - Expert systems are increasingly being used to provide comprehensive interpretative services for diagnosis and treatment planning. Some of these systems are constrained by the complexities of rule-based strategies and a need for knowledge engineers throughout the maintenance phase. A new approach to knowledge acquisition known as Ripple-Down-Rules was used in the development of EICO-1 (Expert Interpretation in Clinical Orthodontics). This expert-maintained system for automating orthodontic reports has a knowledge base of six hundred and eighty rules, and is maintained by an expert trained only in Orthodontics and without the help of knowledge engineers. EICO-1 is the first expert system in Dentistry to use Ripple-Down-Rules. It has potential as an interactive advisory tool and is applicable in a clinical situation. PMID- 11933357 TI - The effect of orthodontic treatment on salivary flow, pH, buffer capacity, and levels of mutans streptococci and lactobacilli. AB - Fixed orthodontic treatment has been shown to cause an increased incidence of enamel demineralisation. The purpose of this study was to investigate the changes in stimulated salivary flow rate, pH, buffer capacity, and the levels of mutans streptococci (MS) and lactobacilli in patients undergoing therapy with fixed orthodontic appliances. Saliva samples, plaque index scores and dietary histories were taken from 21 sequential patients before the start of treatment, and one month and three months after placement of brackets and bands. There was a statistically significant increase in stimulated salivary flow rate, pH, buffer capacity, plaque index scores, and in the levels of MS and lactobacilli after three months of active treatment. It is postulated that the balance between the cariogenic challenge posed by high levels of MS and lactobacilli, and the reparative effects of concurrent increases in salivary flow rate, pH and buffer capacity, determines the likelihood of mineral loss or gain over time. A failure to follow basic preventive measures may increase the risk for some patients of enamel decalcification during orthodontic treatment with fixed appliances. PMID- 11933358 TI - Bond strengths of orthodontic brackets to restorative resin composite surfaces. AB - In orthodontic practice, it is not uncommon to bond brackets to resin composite restorations. With this in mind, this study was designed to compare first the shear/peel strengths of metal, ceramic and polycarbonate brackets bonded to microfilled resin composite (RC), using either a light-cured resin-modified glass ionomer cement (Fuji Ortho LC), a chemical-cured composite (System 1+) or a light cured composite adhesive (Transbond XT); and then to examine the effects of thermocycling on the shear/peel strengths of these systems. Four different brackets were used: two stainless steel (Victory and Optimesh), one ceramic (Transcend 6000) and one polycarbonate (Spirit MB). Seventy-two specimens of each bracket were divided into three groups for bonding with one of the three adhesives. Half the specimens from each group were also thermocycled. Mean shear/peel bond strengths were found to be significantly different for the four different brackets, although not influenced by the three adhesives used within each group. All groups were found to have clinically-acceptable mean bond strengths, except for Spirit MB-System 1+. After thermocycling, both Optimesh Transbond XT and Victory-System 1+ groups showed superior mean bond strengths (26.8 and 24.4 MPa, respectively) when compared with all other groups (p < 0.05). Applying the Weibull survival analysis for groups utilising Victory, Transcend 6000 and Spirit MB brackets, those with 90 per cent or greater probabilities of survival included Victory-System 1+, Transcend 6000-Fuji Ortho LC, Victory-Fuji Ortho LC and Spirit MB-Transbond XT groups. In all groups, bond failure was mainly (64 per cent) cohesive within the RC restorative surface. The thermocycled Spirit MB-Transbond XT group had the highest frequency of undamaged RC failure interfaces. Despite the focus of this study being on bond strength and the potential for surface damage, it was noted that these properties should always be considered alongside other factors such as the strength of the bracket itself, friction within the bracket slot, patients' wishes, cost of the materials and the presenting malocclusion. PMID- 11933359 TI - Tooth movement and vascularity of the dental pulp: a pilot study. AB - The effect of orthodontic tooth movement on the dental pulp was assessed histologically in twelve subjects. The participants in this study required the extraction of at least two maxillary first premolars for orthodontic treatment. They were asked to wear a maxillary removable appliance that acted to move a randomly determined premolar in a buccal direction. The appliance was designed to avoid contacting the contra-lateral tooth that was used as the matched control. The appliance was initially worn for a week to ensure patient comfort and cooperation. The appliance was then activated and the patient dismissed. After two weeks, the appliance was reactivated. Both the control and experimental teeth were extracted three weeks later, on the thirty-fifth day of activated appliance wear. The teeth were fixed, decalcified and sectioned. The sections were stained with haematoxylin and eosin for histological examination. This investigation demonstrated that orthodontic tooth movement did have an effect upon the dental pulp, causing vasodilation in the pulp of an orthodontically stressed tooth. PMID- 11933360 TI - Comprehensive treatment commenced in the mixed dentition and completed in the permanent dentition: an "early treatment" malocclusion. AB - The management of a ten-year-old female with an Angle Class II division 1 malocclusion is followed for nearly ten years, through active treatment, until follow-up some years later. Note is made of: the duration of the overall active treatment period; the need for considerable patient cooperation; the fact that the lower incisors did not move forward on their underlying bone while still moving forward in relation to the APo line; and the fact that, in this instance, the third molars were chosen as appropriate teeth for extraction. PMID- 11933361 TI - Give us evidence that informs the practitioner. PMID- 11933362 TI - The responsibility is yours. PMID- 11933363 TI - A comparison of three primary non-adherent dressings applied to hand surgery wounds. AB - This study reports the results of a prospective randomised controlled trial that compared three non-adherent wound dressings applied to hand surgery wounds. Paraffin-impregnated gauze (Jelonet) was compared with a cellulose, acetate fibre dressing coated with a petrolatum emulsion (Adaptic) and a polyamide net dressing impregnated with silicone gel (Mepitel). The dressings were assessed for their ease of application and removal, pain on removal and wound appearance. Results from 99 patients were available for analysis. Adaptic was significantly easier to remove (p < 0.01), required less soaking (p < 0.05), was less painful to remove (p < 0.05) and caused less wound maceration (p < 0.05) than Jelonet, but was significantly more difficult to apply (p < 0.05). Mepitel was also easier to remove but this did not reach statistical significance. It was also more difficult to apply than Jelonet (p < 0.05). We recommend that Adaptic should be used routinely as the non-adherent dressing for incisions or traumatic wounds on the hand. The slight increased difficulty in applying the dressing is outweighed by the major advantages associated with its removal. PMID- 11933364 TI - The SIGN guideline on the care of chronic leg ulcers: an aid to improving practice. PMID- 11933365 TI - Nurses' views about pain and trauma at dressing changes: results of a national survey. AB - Research findings and professional concerns that patients still experience pain and tissue trauma at dressing changes led specialist nurses to develop a postal questionnaire to elicit the views of 1000 nurses with an interest in wound management. The questionnaire asked practitioners to identify the primary considerations underpinning their approach to pain and tissue trauma at dressing changes, the strategies they adopted, and the factors which determined their treatment choices. The higher than anticipated response rate (37%) illustrates the importance nurses attach to this aspect of practice. The practitioners' main considerations during dressing changes were the need to prevent trauma to the wound (47%) and to avoid causing the patient pain (34%). Significantly, 81% noticed that patients experienced most pain when dressing products were removed, highlighting the need to re-emphasise the principles of moist wound healing. A total of 225 respondents (60%) were aware of dressings specifically designed to prevent pain and trauma during dressing changes. Practitioners also had the freedom to use the product of choice all (61%) or most (36%) of the time. However, respondents also demonstrated confusion about the properties and availability of many dressings. Recommendations for practice include reviewing the number of dressings currently available, together with ongoing education and update on all wound management dressing products and related issues. PMID- 11933366 TI - Doppler assessment. PMID- 11933367 TI - Influences on patient adherence with compression hosiery. PMID- 11933368 TI - Investigating the bacterial barrier properties of four contemporary wound dressings. AB - The ability of four wound dressings (CombiDERM, Allevyn Hydrocellular, Tegaderm and Tielle) to resist penetration of methicillin-resistant Staphylococcus aureus (MRSA), vancomycin-resistant Enterococcus faecalis and Pseudomonas aeruginosa was investigated in vitro using a dedicated test apparatus. With the exception of Tielle, each dressing prevented bacterial transmission over an 11-day challenge period. When both the wound contact surface and the external surface of Tielle were directly challenged with a bacterial suspension, penetration of the dressing was observed within three to five days. The breakdown of its outer membrane could explain the inability of Tielle to maintain a barrier to the passage of the test bacteria used in this wound model. On the basis of these data, CombiDERM, Allevyn Hydrocellular and Tegaderm dressings may facilitate infection control by acting as a physical barrier to the transmission of potentially pathogenic and antibiotic-resistant wound bacteria. However, further research is urgently required to determine whether or not the same results are observed in clinical practice. PMID- 11933369 TI - Can traumacel be used in the treatment of chronic wounds? AB - Traumacel (calcium salt of oxidised cellulose) has previously only been used in the treatment of acute wounds. To assess its safety and effectiveness in the management of chronic wounds a 12-week pilot study was undertaken which involved 11 patients with 15 non-healing leg ulcers and assessed ulcer size, ulcer pain and degree of exudate. Five ulcers healed within the study period. Significant pain relief was experienced by three patients. Use of the dressing did not appear to be related to a reduction in exudate. No patients experienced sensitisation to the product or had to be withdrawn because of adverse effects. Traumacel was found to be safe in the management of chronic ulceration, and appeared to promote healing in some recalcitrant ulcers. PMID- 11933370 TI - Update on stroke prevention and initial acute stroke management. PMID- 11933371 TI - Approach to the patient with polyneuropathy. PMID- 11933372 TI - Headache evaluation and management. AB - Headache is one of the most common complaints in primary care, virtually affecting all persons at some point. Headache may be a symptom of other pathology (secondary headache) or due to a primary headache disorder. Tension headaches, migraines, and cluster headaches are three primary headache syndromes, in order of most frequently occurring to least. The key to effective management of these chronic syndromes is differential diagnosis through history, physical exam, and possibly diagnostic studies. Pharmacologic management may include abortive, adjunctive, and prophylactice medications. PMID- 11933373 TI - Understanding Parkinson's disease: detection and early disease management. AB - Parkinson's disease (PD) is a common, debilitating, neurodegenerative disorder characterized by neuronal loss within the basal ganglia and insufficient levels of the neurotransmitter dopamine. Symptoms include resting tremor, rigidity, bradykinesia (slowness of voluntary movement), and postural disturbances. Exact cause is unknown, but theories surrounding environmental or endogenous toxicities have been suggested. Differential diagnoses include genetic and other neurologic disorders that may share symptoms similar to those seen in PD. Clinical progression has been categorized into three phases of the disease: early, nonfluctuating, and fluctuating. Medications generally offer good symptom relief during the early and nonfluctuating phases of the disease. Classifications of anti-PD medications include anticholinergics, dopamine agonists, amantadine, MAO B inhibitors, levodopa-carbidopa, and Catechol-o-methyl transferase inhibitors. Surgical intervention may be an option for select patients whose conditions are not well controlled though medical management strategies. Primary care providers often can manage patients in the early stage of PD, but later stages require expert neurologic management. Patient/family education and anticipatory guidance is imperative. PMID- 11933374 TI - Seizure disorders and epilepsy. AB - Seizures are uncontrolled hypersynchronous electrical discharges of neurons in the brain that interfere with normal function. They are a symptom of an underlying disorder. Epilepsy is a condition of recurring seizures that do not have a reversible metabolic cause. Seizures can be confused with a variety of other conditions, so an understanding of seizure manifestations is crucial in making an accurate diagnosis. Drug therapy is the mainstay of epilepsy treatment, but surgery and vagal nerve stimulation are options for selected refractory cases. Psychosocial consequences of recurring seizures are often more significant to patients than the seizures themselves. PMID- 11933375 TI - Medications used to prevent adhesion and clotting. PMID- 11933376 TI - Patient with memory loss. PMID- 11933377 TI - Carpal tunnel syndrome. PMID- 11933378 TI - Polyneuropathy. PMID- 11933379 TI - Clinton threatening to veto Medicare bill with composite rate increase, lifetime transplant meds. PMID- 11933380 TI - Is this a police state or what? PMID- 11933381 TI - Hemolytic-uremic syndrome associated with Escherichia coli 0157:H7. PMID- 11933382 TI - What are the next steps for improving oversight in renal care? Tougher conditions for coverage may improve standards of care. PMID- 11933383 TI - Missed treatments: how to help patients change problem behavior. PMID- 11933384 TI - Managing the Achille's heel of dialysis. PMID- 11933385 TI - Can the type of access influence outcomes? PMID- 11933386 TI - Using CQI to improve vascular access management in the dialysis unit. PMID- 11933387 TI - A review of vascular access management in Europe. PMID- 11933388 TI - CPM project shows catheters deliver less dialysis. PMID- 11933389 TI - Biolink, Vasca push new access devices. PMID- 11933390 TI - Preliminary findings from the National Pre-ESRD Education Initiative. AB - Regardless of age or race, more patients preferred peritoneal dialysis (PD) than would be expected from the current ratio of patients treated with PD in the U.S. The percent of patients choosing PD seems to be largely influenced by the scope of information they receive prior to therapy initiation. In the absence of thorough treatment option education, a significant number of patients may be assigned a therapy when, in reality, they would have selected an alternative option. There is a need to further understand the correlation between a practice's PD utilization rate and the extent to which patients are given dialysis treatment options. PMID- 11933391 TI - Canadian study uses controls to show benefits of daily dialysis. PMID- 11933392 TI - Amendment proposed for funding daily hemodialysis. PMID- 11933393 TI - New CDC surveillance network tracks vascular access infections. Plus: early data from annual facility surveys show increases in catheters, fistulas. PMID- 11933394 TI - Cases of acute viral hepatitis increasing, but hepatitis B declining. PMID- 11933395 TI - Body mass may not be good predictor of dialysis outcome. PMID- 11933397 TI - NIH: ACE inhibitors for HTN protect kidneys. PMID- 11933398 TI - Baby made to order for stem cell transplant. PMID- 11933396 TI - NESP researchers present latest data at ASN. PMID- 11933399 TI - St. John's Wort linked to kidney rejection. PMID- 11933400 TI - Negotiating the boundaries: the experience of the mental health nurse at the interface with the criminal justice system. AB - Community mental health nurses working within the criminal justice system undertake an important function with regard to the strategic intent that wherever possible, mentally disordered persons should receive care and treatment from health and social services. Their precise role, and the particular function of court diversion/assessment schemes, is still being defined. This takes place within a broader context of reorientation of mental health nursing within the move to community care and exposure to the complexity of multidisciplinary working. Examining the experience of mental health nurses in court diversion schemes reveals the intensity of the experience of working in an inter professional setting without the benefit of preparation or training, and without the frame of reference of a structured health environment. Adaptation of the language of mental health nursing was necessary to effect mutual understanding with other professional groups, and there was a recognition that the competences needed to operate in this environment had not previously been identified as part of their skills base. The result was to bring into question their professional practice and identity as mental health nurses. The handling of patient information in the context of an inter-professional setting remains an issue requiring clarification. PMID- 11933401 TI - An inquiry into moral virtues, especially compassion, in psychiatric nurses: findings from a Delphi study. AB - A three-round Delphi study was conducted to gather data on ethical reasoning among psychiatric nurses (N = 26 in round one (R1), decreasing to N = 14 in the final round (R3)). Transcripts of questionnaires were carefully read and compared. Responses were manually sorted into categories, themes and patterns of interest. Eight debates emerged from the data. This article discusses two in detail: the nature of moral virtues and the meaning of compassion in psychiatric nursing. A sympathetic overview of virtue ethics is also provided. The nurses' responses included a lot of virtue terms, such as, 'honest', 'fair', and 'care". However, 'nurses' moral virtues' was ranked low in importance as a notion invoked in ethical decision making in the round-one ranking exercise. Only half of the sample believed that the moral character of a psychiatric nurse is important in ethical decision-making. Further, most of the round-one sample thought the virtues could not be acquired. Compassion was identified as crucial to psychiatric nursing and the nurse-client relationship, though, as expected, many diverse meanings were attributed to this notion. While the Delphi method proved adequate for our purposes, problems with regard to accurately understanding the respondents' intended meanings highlighted a major weakness of this technique, in common with other methods relying on questionnaires. Further inquiry is needed regarding the role of moral virtues and virtue ethics in both psychiatric nursing and nurse education. PMID- 11933402 TI - 'Partnership': a co-operative inquiry between community mental health nurses and their clients. 1. Research methodology, process and reflections. AB - The aim of this study was to explore the utility of co-operative enquiry between community mental health nurses and their clients. The research design was a form of co-operative enquiry. Two nurses and two clients volunteered to participate as co-researchers and co-subjects with two facilitators in a co-operative inquiry group. The subject of the inquiry, agreed by the group, was the relationship between the nurse and client. The method for the inquiry was developed by the participants and consisted of dialogue groups. This paper provides a description of the research design and methodology with a first-person account of the research experience and reflections by the two facilitators. The process is considered with particular reference to the influence of the study facilitators. Their position changes from wanting to influence an 'external' relationship to one of being influenced within the research relationship over time through reflection and growing self-awareness. This results in the inquiry ultimately being driven by the agenda of the group rather than the process. The outcomes of the study were both informative and transformative and the content of the inquiry relating to the nurse-client relationship will be presented in a second paper. PMID- 11933403 TI - Self-esteem in community mental health nurses: findings from the all-Wales stress study. AB - The authors conducted an all-Wales survey of community mental health nurses (CMHNs) to determine their levels of stress, coping and burnout. A total of 301 CMHNs were surveyed in 10 NHS Trusts in Wales. A range of measures were used. These included the General Health Questionnaire (GHQ-12), Maslach Burnout Inventory (MBI), Rosenberg Self-Esteem Scale (SES), Community Psychiatric Nursing (CPN) Stress Questionnaire, and PsychNurse Methods of Coping Questionnaire. The findings from the Rosenberg SES are reported here. Community mental health nurses in Wales scored as having average self-esteem. When the data were divided into high and low self-esteem, a large group of CMHNs (40%) were found to have low self-esteem. Factors that are associated with low and high self-esteem were identified. Alcohol consumption and being on lower nursing grades (D, E, F) were associated with low self-esteem, whilst amount of experience working as a CMHN was associated with high self-esteem. PMID- 11933404 TI - Psychiatric nurses' lived experiences of working with inpatient care on a general team psychiatric ward. AB - To reveal the meaning of being a nurse working with inpatient care on a team psychiatric ward in Sweden, 22 psychiatric nurses were interviewed and the transcribed texts were analysed by means of latent content analysis. Three themes emerged from the analysis: developing a working relationship with the patient in everyday caregiving; encountering and handling the unforseeable in daily living; and struggling with professional independence and dependency. Developing a working relationship with the patient in everyday caregiving meant that the nurse patient relationship was the foundation of the caregiving and included being with, as well as doing for, and with, the patient. Four different approaches in daily caregiving were revealed: networking, teaching, containing and protecting. The nurses' approaches in the nurse-patient relationship alternated between being an 'expert' and a 'collaborator'. Encountering and handling unforeseeable situations meant that the nurses were exposed to and had to be prepared for unpredictable situations where they were on their own, handling sometimes strong emotional reactions and relying on their own ability to act. Struggling with professional independence and dependency meant that the nurses seemed to lack professional confidence, although they had many responsibilities, but also less authority to decide about overall care planning. Contextual aspects such as organizational hindrance, unsatisfactory work-environment and co-operation difficulties were illuminated. The result indicates the need for a stable and predictable organizational structure if nurses are to manage the demanding nurse patient relationships that everyday caregiving requires. A question highlighted by this study is whether multidisciplinary team organization has been effectively developed in Sweden, as uncertainty about the roles and responsibilities of nurses was apparent. PMID- 11933406 TI - The strengths and weaknesses of cognitive behavioural approaches to treating depression and their potential for wider utilization by mental health nurses. AB - Depression is widely acknowledged to be the major factor implicated with suicide, an enormous financial cost on the economies of western countries and a source of intense despair for millions of people around the world. A steady stream of articles are published both in popular, generic and specialist nursing journals that illustrate the potential of cognitive behavioural therapies in the treatment of depression. Should these therapies be restricted for use by registered therapists or do the techniques have a wider application? The marketing of these approaches for use by nurses prompted a review of the purported strengths and weaknesses of these approaches in comparison with other possible alternatives. Many mental health nurses in community settings already use cognitive behavioural approaches with clients. Here it will be argued that several of the recognized strengths of cognitive behavioural approaches lend themselves to incorporation in nurse-patient interactions in varied in-patient settings by nurses who spend protracted periods of time with depressed patients but lack formal therapist qualifications and do not consider themselves counsellors. PMID- 11933405 TI - Nurse behavioural psychotherapy and pathological gambling: an Australian perspective. AB - Nurses have been employed as behavioural psychotherapists for over 25 years. The first training in this field was established at The Maudsley Hospital (Marks et al. 1977). Since then there have been a number of courses producing many specialist nurse behavioural psychotherapists (NBTs) (Duggan et al. 1993). Nurse behavioural therapy is taken to mean nurses working as behavioural psychotherapists. This title does not mean they are practising a separate form of therapy from other professionals such as psychologists. In Australia there is no such training, with nurse behavioural psychotherapists working here having trained first in UK. Initially NBTs were trained to treat people with anxiety disorders. Over the past 25 years this emphasis has remained, but many NBTs have gone on to provide help to people with other problems. This paper will demonstrate the practical application of behavioural psychotherapy with problem gamblers, and how specialist trained nurses can provide such treatment. The paper will look briefly at nurse behavioural psychotherapy from its initial inception to its current status in Australia. The outcomes of the treatment programme carried out at Flinders Medical Centre for Problem Gamblers will be discussed with a view to nurses carrying out therapy. Finally, the future of nurse behavioural psychotherapy in Australia and how nurses can apply such skills across a wide range of problems, including gambling, will be reviewed. Aspects of this paper were first presented to the 7th NAGS conference, Adelaide 1996. PMID- 11933407 TI - Lessons from history: the politics of psychiatry in the USSR. AB - The political-economic base of society affects every aspect of it, including nursing and psychiatry. This can be demonstrated by making a historical analysis of societies with different political-economic systems. Psychiatry in the USSR took a different form to psychiatry in the West. Differences included the diagnostic categories used and treatments employed. This can be investigated by examining accounts of clinical practice. Soviet psychiatry was also used for the systematic incarceration of political dissidents. Some commentators have drawn on the Soviet experience and used it to support an argument that psychiatry operates as a form of social control in the West as well as the USSR. This article shows how psychiatric abuse in the USSR was a historically specific response to a particular situation. Therefore some of the conclusions about Western psychiatry extrapolated from the Soviet experience are unsupportable. Whatever the role of psychiatry in the West, its mechanism is qualitatively different to that which existed in the USSR. In order to understand why Soviet medical workers were co opted into the conscious abuse of psychiatry, it is essential to understand the specific nature of the USSR. This does not necessarily allow generalizations about Western psychiatry to be made from the Soviet experience. As psychiatric nurses, we can also learn from a particularly tragic period of psychiatry's history. PMID- 11933408 TI - One law for the lion and the ox.... A reply to Burnard and Hannigan. PMID- 11933410 TI - Qualitative and quantitative research: a question of synthesis? PMID- 11933409 TI - Synthesis and sterility: a reply to Burnard and Hannigan. PMID- 11933411 TI - Necessary debates: the need for controversy in mental health. PMID- 11933412 TI - An application of conjoint analysis to the process of psychiatric day hospital care. PMID- 11933413 TI - Nurse consultants must put holistic care first. PMID- 11933414 TI - The physics of sub-bandage pressure measurement. AB - Compression therapy is the principal treatment for leg ulcers associated with venous disease. The extent of compression generally can be estimated from the general Laplace equation relating pressure, bandage tension and leg radius. This study aimed to investigate the physical forces under long stretch bandages applied under constant tensions to cylinders of three defined radii ('model limbs'). Force expressed as pressure was measured under one, two and three layers of eight long stretch bandages and a plastic film (control) using temperature compensated, strain-gauge pressure transducers. The bandages were applied at the manufactures' recommended extensions/tensions and at constant tensions (2-10N) to three rigid plastic tubes of increasing size, covered with a layer of soft foam. Foams of different hardnesses ranging from 20 to 230N were assessed. The median applied pressure and observed bandage characteristics changed with bandage type. The median applied pressure also changed with tension, number of layers, tube radius and foam hardness. The inter-layer shear component also tended to increase with the number of layers. An increase in sub-bandage pressure was observed relative to the radius, tension, foam hardness and number of layers but the relationship was not directly proportional. The shear force tended to be associated with increased bandage layers. PMID- 11933415 TI - Assessment of infection in diabetic foot ulcers. AB - Typical signs of infection in diabetic foot ulcers are often absent or late. This literature review outlines the factors practitioners must take into account when assessing for and managing such infections. PMID- 11933416 TI - Effect of four treatment variants on the functional and cosmetic state of mature scars. AB - A randomised, intra-individual, comparative study demonstrated that both qualitative improvements and significant changes in skin functional condition can be achieved in the tissue of older, mature scars. Four treatment modalities were studied in an intra-individual comparison involving 12 volunteers with 2.5 to 4 year-old scars. The treatments were: a self-adherent, hydroactive, polyurethane dressing alone; polyurethane plus compression; silicone sheeting plus compression; and compression alone. Evaluation criteria were changes in the microcirculation, roughness and the skin temperature of the scar tissue. All treatment modalities were found to have significant effects both on tissue function and scar tissue surface structure. The most pronounced effects were achieved with the combination of polyurethane dressing plus compression or silicone sheeting plus compression. The positive effect of the polyurethane dressing alone on scar tissue was even slightly superior to that of compression therapy alone. PMID- 11933417 TI - Pressure-ulcer prevention strategies: are the data valid? PMID- 11933418 TI - Use of adjunctive therapy to heal a diabetic amputation site. AB - Diabetes can result in a range of complications, and is responsible for many lower limb amputations. This case report discusses a diabetic patient with a complex, non-healing wound following amputation of the foot. A number of adjunctive therapies were used to promote healing, and led to the complete healing of the wound. PMID- 11933419 TI - Paste bandages and chronic wound cavities. PMID- 11933420 TI - The impact of minor acute wounds on quality of life. AB - A study was undertaken to explore the experiences of patients with minor acute wounds following discharge from either inpatient or day care. The sample comprised patients attending the plastic surgery dressing clinic of a district general hospital. Patients whose wounds were considered to be chronic were not included. A conceptual framework was developed from the literature, postulating that the patient is 'eclipsed' by the wound and enters the sick role before healing and recovery take place. A self-administered questionnaire was used to collect data for a mainly quantitative analysis, with some open questions to indicate avenues for further study. Unfortunately, the response rate was low, and insufficient data were obtained for effective analysis. However, trends were identified which indicated that these wounds had a significant impact on patients and their families, and recommendations were made for future studies. The survey would need to be repeated on a larger, more representative sample and include semi-structured interviews to produce generalizable results. PMID- 11933421 TI - Assessing nurses' knowledge of wound management. AB - An understanding of the principles and theories of wound healing and management is vital for the delivery of high-quality care. This literature review suggests that wound management is often undertaken by health-care professionals who have not received sufficient training in this specialty. PMID- 11933422 TI - RV dialysis: a dream realized and a dream threatened. PMID- 11933423 TI - Monitoring quality in the dialysis unit: is it enough? GAO, OIG reports looks for government scrutiny, find little; is the fox guarding the hen house? PMID- 11933424 TI - Improving donation in Europe: the Transplant Procurement Management (TPM) project. PMID- 11933425 TI - NEJM study shows danger of calcium-laced supplements for ESRD patients. PMID- 11933426 TI - Sodium, not fluid, controls interdialytic weight gain. PMID- 11933427 TI - Part I. On bended knee: how dialysis patients passed an ESRD licensure bill in Georgia. PMID- 11933429 TI - Rumors continue on Baxter/Gambro merger. PMID- 11933428 TI - HCFA releases proposed changes to 2001 Medicare fee schedule. PMID- 11933430 TI - Gambro settles Michigan anti-trust case, lab charges. PMID- 11933431 TI - National Transplantation Pregnancy Registry looks at outcomes with Neoral and Tacrolimus. PMID- 11933432 TI - Sangstat recalls SangCya. New-found study shows biocompatibility problems with Neoral. PMID- 11933433 TI - Generic substitution of immunosuppressant medications in organ transplantation. PMID- 11933434 TI - "Transplant 2000" plays host to important breakthroughs, new drug studies. PMID- 11933435 TI - Can Rapamune replace cyclosporine? No significant difference in rejection rate after six months; long-term studies needed. PMID- 11933436 TI - Chronic nasal obstruction: allergy or infection? PMID- 11933437 TI - Acute and chronic pharyngitis across the lifespan. AB - Although most instances of sore throat are caused by relatively benign infectious or noninfectious processes, pharyngitis may herald serious or even fatal illnesses. Viral pharyngitis is the diagnosis in most cases, but because GABHS is the most common bacterial organism requiring antimicrobial treatment, an office visit is often necessary. There is no exact constellation of signs and symptoms that is pathognomonic for GABHS; nevertheless, sudden onset of sore throat with fever and cervical lymph node tenderness, in the absence of cough and nasal symptoms, is at least suggestive in adults, and possibly in children. Physical examination and prudent use of laboratory testing will assist in the diagnosis of both acute and chronic pharyngitis. The primary care provider who promptly identifies and properly treats patients infected with S. pyogenes has reduced the number of missed school or work days, the risk of developing ARF, the likelihood of transmission to others, and inappropriate use of antibiotics for those with other causes of sore throat. Further education of patient, family, and other clinicians will reduce medical expenses, avoid unnecessary antibiotic exposure, and inform the public regarding judicious management of pharyngitis. PMID- 11933438 TI - The evaluation of the client with acute bacterial rhinosinusitis. AB - The diagnosis of acute bacterial rhinosinusitis (ABR) remains elusive. Most patients with symptoms of ABR are seen in primary care offices. This article focuses on the presentation of the patient with symptoms of ABR, accurate diagnosis, and pharmacologic treatment. The article also discusses the appropriate referrals for patients who do not respond to conventional treatment. PMID- 11933439 TI - Epistaxis management: what's new and what's noteworthy. AB - In many ways, the treatment of epistaxis is not new. Nasal packing was used in the 4th century BC. Our modern-day management of epistaxis is a little more sophisticated, with the advent of new products, rigid endoscopes, improved surgical techniques, and arterial embolization. Research plays a critical role in shaping our epistaxis practice management. Epistaxis is recognized as one of the most common ear, nose, and throat (ENT) problems; it affects persons of all ages. Proper first aid steps can be effective. Only 10% of individuals with nasal bleeding seek medical attention. Health care providers in all primary care settings encounter patients experiencing epistaxis and need to be knowledgeable in emergency epistaxis management. The evaluation will be key to successful identification of the site of bleeding and to identify possible underlying cause(s) that will guide treatment options. PMID- 11933440 TI - Tobacco smoke exposure and pediatric otitis media: an empirical basis for practice. AB - Tobacco smoke exposure has been attributed to a myriad of pediatric clinical outcomes, with a staggering financial burden to society. This article is an examination of published research that has focused on the relationship between tobacco smoke exposure and pediatric otitis media. The weight of evidence presented supports a positive relationship between the two clinical entities. Further clinical studies are needed so primary care providers may continue to build a strong empirical base for interventions directed at helping parents reduce tobacco smoke exposure in their children's home environments. Interventions based on the Ask, Advise, and Assist framework are recommended. PMID- 11933442 TI - Young boy presenting with swollen glands and sore throat. PMID- 11933441 TI - Antimicrobial choices in the treatment of acute otitis media. PMID- 11933443 TI - Acute otitis externa. PMID- 11933444 TI - Acute laryngitis. PMID- 11933445 TI - Nasal sprays and irrigations. PMID- 11933446 TI - Ear wax. PMID- 11933447 TI - Cold or flu. PMID- 11933448 TI - Communication matters. PMID- 11933449 TI - Effect of visco-elastic foam mattresses on the development of pressure ulcers in patients with hip fractures. AB - This study had three aims: to investigate if visco-elastic foam mattresses are more effective than standard hospital mattresses in reducing the incidence of pressure ulcers in patients with hip fractures; to compare pressure ulcer grade and location and documented nursing prevention and treatment interventions in patients using the two types of mattresses; to identify possible predictors of pressure ulcer development. Using a prospective randomised controlled trial design 101 patients (mean age: 84 years) were randomly allocated either a visco elastic foam mattress or a standard mattress. There was no significant difference in the incidence of pressure ulcers between the two groups, but patients on standard mattresses tended to develop more severe pressure ulcers. Furthermore, according to the documentation, patients with grade I pressure ulcers who were allocated a standard mattress received more preventive interventions, which may have reduced the differences in outcomes between the two groups. The researchers concluded that the results support the use of the test mattress. Significant predictors of pressure ulcer development were long waiting times for surgery and low haemoglobin levels at hospital admission. PMID- 11933451 TI - When is reduced-compression bandaging safe and effective? AB - During a one-year period 44 patients were treated with reduced-compression bandaging in the vascular unit at a district general hospital. Twenty patients with venous ulcers were referred from community leg ulcer clinics because they could not tolerate standard compression bandaging (group 1). A further 24 patients had mixed aetiology leg ulcers (group 2), and high-compression bandaging was not attempted because of significant peripheral vascular disease (ankle brachial pressure index < 0.8). Forty-two patients tolerated reduced-compression bandaging without discomfort or skin pressure changes. Healing was achieved in 32 patients (13 in group 1 and 19 in group 2). All non-healed ulcers were considered smaller at the time of final follow-up, and had less exudate. Reduced-compression bandaging has been shown to be effective and well tolerated by patients with venous ulcers who cannot comply with high-compression bandaging. With close supervision, it is safe to use in patients with significant peripheral vascular disease. PMID- 11933450 TI - Using an esterified hyaluronan fleece to promote healing in difficult-to-treat wounds. PMID- 11933452 TI - Use of iodine in a dehisced thoracotomy wound. PMID- 11933453 TI - How effective is the Abbreviated Burn Severity Index in predicting patient mortality? PMID- 11933454 TI - Developing skills in leg ulcer nursing: the lessons learned. AB - The high treatment costs of inappropriate leg ulcer management are well referenced. Wide variations in practice have also been reported. A training programme for community nurses has been in place in Leicestershire and Rutland NHS Trust for five years. Its objective is to promote evidence-based leg ulcer management. The aim of this prospective audit was to assess and compare the effectiveness of the compression bandage systems used in the trust: the original Charing Cross system (August 1996 to September 1997) and Robinson's Ultra Four kit (October 1997 to December 1998). All patients with leg ulceration were assessed using the Leicestershire leg ulcer assessment form. This was submitted along with an audit tool to the tissue viability service in exchange for a 12 week supply of compression bandages. The audit form was resubmitted after each subsequent 12-week period until the patient either had healed or was withdrawn from the audit. Differences between the two groups in mobility, previous deep vein thrombosis, fixed ankle deformity, the male to female ratio, duration of ulcer and ulcer surface area were noted. Patients using the Ultra Four kit had faster healing times than those given the Charing Cross system, but this was not statistically significant. Cost comparisons between the previous dressing regimen and compression bandaging showed a significance difference (p < 0.001). There was also a significant reduction in nurse time (p < 0.001). The audit results showed that compression bandaging therapy was an effective method of achieving healing in patients with venous leg ulcers and that significant cost savings were made in terms of dressing cost and nurse time. Furthermore, there were potential benefits in continuing compression therapy for the group of non-healers. PMID- 11933455 TI - Measuring sub-bandage pressure. AB - Compression therapy is the principal treatment for leg ulcers associated with venous disease. The extent of compression generally can be estimated from the general Laplace equation relating pressure, bandage tension and leg radius. This study aimed to investigate the physical forces under long-stretch bandages applied under constant tensions to cylinders of three defined radii ('model limbs'). Force expressed as pressure was measured under one, two and three layers of eight long-stretch bandages and a plastic film (control) using temperature compensated, strain-gauge pressure transducers. The bandages were applied at the manufactures' recommended extensions/tensions and at constant tensions (2-10 N) to three rigid plastic tubes of increasing size, covered with a layer of soft foam. Foams of different hardnesses ranging from 20 to 230 N were assessed. The median applied pressure and observed bandage characteristics changed with bandage type. The median applied pressure also changed with tension, number of layers, tube radius and foam hardness. The inter-layer shear component also tended to increase with the number of layers. An increase in sub-bandage pressure was observed relative to the radius, tension, foam hardness and number of layers but the relationship was not directly proportional. The shear force tended to be associated with increased bandage layers. PMID- 11933456 TI - [Dipeptidyl(amino)peptidase IV in the differential diagnosis of thyroid gland tumors: methods and results of a pilot study of 200 cases]. AB - The use of dipeptidyl aminopeptidase IV (DPP IV) staining by azo-coupling in preoperative and intraoperative diagnostics of thyroid lesions is presented. In a series of 200 histologically confirmed cases examined, the sensitivity and the specificity were 71% and 99%, respectively in 124 smears, and 70% and 94%, respectively in 189 frozen sections. DPP IV expression showed high negative predictive value as well. DPP IV is suggested as an additional tool in the preoperative and intraoperative diagnostics of thyroid lesions. PMID- 11933457 TI - [Detection of carriers of Duchenne and Becker muscular dystrophy using the fluorescence in situ hybridization method]. AB - Duchenne and Becker muscular dystrophies (DMD/BMD) are X-linked recessive disorders caused by mutations in the dystrophin gene. A large intragenic deletion has been described in about 65% of DMD/BMD patients. Mothers of affected males are DMD/BMD carriers in two thirds of the cases. Routine deletions detection in DMD/BMD males is performed using multiplex polymerase chain reaction (mPCR), RT PCR with a protein truncation test (PTT) or using Southern blotting. In females the deletions detection is complicated by the presence of a normal gene copy on the second X-chromosome. We are presenting the diagnostic strategy using FISH for the deletions detection in the dystrophin gene of female DMD/BMD carriers. We have used a set of six cosmid probes for the detection of the most frequently deleted areas of the dystrophin gene from the Department of Human Genetics, Leiden University Medical Center. We have examined 14 mothers of DMD/BMD males with a deletion in the dystrophin gene identified using mPCR. Four mothers of affected males have been diagnosed as carriers of a deletion in the dystrophin gene. We have revealed no deletion mutations in the exons examined in a control group of four healthy females. No discrepancy has been found between the FISH analysis results and the results of mPCR. Our results indicate that FISH is an effective and direct method for the identification of DMD/BMD carriers and we suggest this method as a method of a first choice in the identification of DMD/BMD carriers. PMID- 11933458 TI - [Lymphoproliferative disease after transplantation]. AB - Herein we describe 7 cases of posttransplantation lymphoproliferative disease (PTLD), 5 in men and 2 in women (aged from 25 to 62 years), occurring from 4 months to 12 years (mean, 7 years) after transplantation. Our patients were recipients of kidney, kidney and pancreas, heart, and autologous peripheral haematopoetic stem cells. Four cases were diagnosed as monomorphic and three as polymorphic type of PTLD according to the WHO classification. Monoclonal immunoglobuline heavy chain gene rearrangement was detected in two monomorphic lesions and one polymorphic lesion by polymerase chain reaction (PCR). In the two cases of polymorphic and the one case of monomorphic PTLD, the presence of EBV was visualised by immunohistochemical staining of some transformed lymphoid cells for latent membrane protein (LMP) of EBV. The presence of type A EBV was demonstrated by PCR. The patients were treated by reduction or discontinuation of immunosuppression and by chemotherapy. In 2 cases, a part of the organ affected by lymphoma (sigmoid colon and pancreas) was surgically resected. Four patients died of causes related to PTLD (2 to 15 months after the diagnosis), mainly of infectious complications. Two other patients who achieved remission died of unrelated causes. Only the youngest man is alive and in the complete remission 10 months after the diagnosis of PTLD. PMID- 11933459 TI - [Syndecan-1 (CD138): an immunohistochemical marker of plasma cell tumors]. AB - The immunohistochemical detection of syndecan-1 (belonging to the cluster CD138) is a sensitive and reliable method for identifying normal and neoplastic plasma cells. It may be used in paraffin-embedded bone marrow specimens, as well as in extramedullary tumours of unknown origin. The three anaplastic tumours reported by us in the lymph node, the gingiva, and pleura were negative for other markers, but the syndecan-1 positivity elucidated their plasmocytic histogenesis. PMID- 11933460 TI - [Pulmonary thromboembolism over a 25-year period in data from the Institute of Pathology of the P.J. Safarik Medical School in Kosice]. AB - Based on a review of archival autopsy protocols an analysis of the frequency of pulmonary thrombembolism in consecutive autopsy material is presented. The study interval for the analysis was oriented to the period between the years 1949-2000. Our analysis has shown a gradual progression of incidence of pulmonary thrombembolism, as demonstrated in relative figures. The applied preventive measures against the development of thrombosis during hospitalization have not substantially contributed to decreasing its frequency, what is not in accord with some recent observations from other countries. PMID- 11933461 TI - [Role of the MIB-1 proliferative markers in the diagnosis and prognosis of tumors of the thyroid gland]. AB - Well-differentiated thyroid tumors may sometimes cause diagnostic uncertainty due to difficulties in the evaluation of certain morphological criteria (capsular and/or vascular invasion, cytomorphological features). Therefore, various diagnostic/prognostic markers are currently studied, namely the markers of tumor proliferation. The aim of our study was to evaluate the proliferative MIB-1 index in 155 thyroid tumors, and to correlate it with morphological diagnosis, size of the tumors, and the patients' age. Oncocytic tumors were represented by 59 follicular adenomas, 27 follicular carcinomas and 12 papillocarcinomas. Nononcocytic tumors comprised 24 follicular adenomas and 33 conventional papillary carcinomas. The Ki-67 antigen (formalin resistant epitope MIB-1) was detected immunohistochemically and the proliferative index (PI) of tumors was evaluated. The results were statistically analyzed using analysis of variance (ANOVA) and Wilcoxon tests (significance level p < 0.05). Carcinomas showed significantly higher PI than adenomas. Moreover, PI in oncocytic adenomas was higher than in nononcocytic ones. However, proliferative activity in all types of the carcinomas was similar. The higher rates of proliferation correlated with the advanced age of the patients with follicular carcinomas (p < 0.0016). PMID- 11933462 TI - Mitochondrial cardiomyopathy--case report. AB - A male infant developed progressive neuromuscular disease, hypertrophic cardiomyopathy and brain atrophy since the birth. Increased level of lactate with increased lactate/pyruvate ratio suggested a disturbance in the mitochondrial energy metabolism. The activities of respiratory chain complexes III, IV and II + III, of pyruvate dehydrogenase complex and of citrate synthase in isolated muscle mitochondria were low in comparison with controls, with parallel decrease in the content of protein amount of respiratory chain complexes III and IV. No large scale deletions of mitochondrial DNA (mtDNA) and mtDNA point mutations A3243G, A8344G or T8993G indicating syndromes MELAS, MERRF or NARP were detected. The boy died at the age of 7 weeks. The autopsy revealed typical changes of mitochondrial cardiomyopathy-marked myocardial hypertrophy with muscle pallor, histological finding of diffuse fine granularity of the cytoplasm in the perinuclear regions, and ultrastructural findings of mitochondrial hyperplasia, enlargement (megamitochondria) and abnormal shape. PMID- 11933463 TI - [Myoepithelial carcinoma in adenomyoepithelioma of the breast (malignant adenomyoepithelioma)--case report]. AB - There is described fifty-year-old woman with so called malignant adenomyoepithelioma of the breast. The conventional benign looking adenomyoepithelioma with biphasic tubular pattern continually fused to unequivocal invasive lesion which by immunophenotype turned out to be myoepithelial carcinoma. The 15 months follow up was uneventful. Extensive literature overview and terminology consideration is presented. The paper discusses biologic consequences of this rare breast lesion and its close resemblance to almost identical lesions in the salivary glands and skin adnexa. PMID- 11933464 TI - The experience of being an adolescent child of a parent who has a mood disorder. AB - The purpose of this phenomenological study was to describe the experiences of adolescent children living with a parent who has a mood disorder. Three female adolescents each participated in two unstructured, tape-recorded interviews. Interviews were analysed using the Giorgi (1985) method. Three themes emerged. First, participants experienced a serious disruption in family life due to the unavailability of the ill parent, which evoked feelings of an intense sense of responsibility, fear and loss. This disruption led to the adolescent taking on parental roles and tasks. Second, participants perceived themselves to lack knowledge and understanding, leading to frustration and fears that their parents may not get better, and that they themselves may become ill like their parents. They sought information in order to better adjust to and understand the illness. Third, participants struggled to come to terms with the burden of having a psychiatrically ill parent. The effort of trying to cope and make sense of the experience involved a variety of cognitive and affective responses. Implications for nursing practice and research are discussed. PMID- 11933465 TI - An evaluation of the implementation of a liaison service in an A&E department. AB - This paper describes the results of an evaluation of a multidisciplinary liaison service based in an A&E department of a district general hospital in South Wales. The aims of the services were to increase the rate of referral for psychosocial assessment of individuals who presented at the department following acts of deliberate self-harm and to increase the number of such assessments completed within 24 h. The paper describes the context in which such a service was developed and outlines how this preliminary evaluation was completed. Data were collected from hospital records, over two corresponding five-month time-periods, in the year before the implementation of the service and the year following implementation of the service. The results show that the implementation of the service led to a statistically significant increase in the rate of referral for assessment by accident and emergency staff. Amongst other positive outcomes, the rate of repeat presentations was reduced for the period of one year following initial presentation. The implications of these results are discussed in relation to other research in the area and the suggestion is made that nursing staff may be more clinically and cost effective in completing psychosocial assessments in A&E departments than medical staff. PMID- 11933466 TI - Self-image and burnout in psychiatric staff. AB - Burnout was tested for in 754 mental health workers and related to self-image as assessed with Structural Analysis of Social Behavior (SASB, Benjamin 1974). A positive relation was found between burnout and negative self-image, and between the experience of personal accomplishment and positive self-image. Compared to self-image, gender, age and work setting did not explain any variance in burnout. Highly burned-out persons had a significantly more negative self-image than staff who had rated themselves as low burnout. Finally, the relation between self-image and burnout was studied in 210 subjects who had completed their self-image ratings one year before burnout was measured, with the same results: a negative self-image was related to higher burnout one year later. One general conclusion is that a tendency in staff to treat themselves in negative ways may function as a negative filter for coping with difficulties at work and thus be a risk factor for burnout. PMID- 11933467 TI - What are the criteria that mentors use to make judgements on the clinical performance of student mental health nurses? An exploratory study of the formal written communication at the end of clinical nursing practice modules. AB - Issues concerning the assessment of student nurse performance in clinical practice have pre-occupied nurse educationalists for some time and continue to pose problems. The debate in the nursing literature tends to revolve around two seemingly mutually exclusive positions related to competence vs. learning outcomes. These are views that seem to be held by academics with little reference to clinical nurses who have responsibility for the assessment of student performance in clinical practice. This paper adds to the wider debate on clinical assessment by reporting on a small research study that explored the written comments that 150 mentors made in relation to the performance of student mental health nurses following periods of clinical practice. An extensive literature review is provided in order to place the debate in context, followed by a brief outline of the research study, its findings, discussion and conclusion. This analysis of findings demonstrates that student learning is of major concern to mentors, and that the assessment of student performance in clinical practice is not restricted by pre-determined behavioural learning outcomes. Personal characteristics of students exert a great influence on judgements about clinical performance. This paper does not offer solutions to the search for the perfect clinical assessment, but it does call for an increased dialogue between educationalists and mentors to discuss the implications of this research for the development of an appropriate clinical assessment tool. PMID- 11933468 TI - Being-with, doing-with: a model of the nurse-client relationship in mental health nursing. AB - The authors describe a model of the nurse-client relationship within mental health nursing. This is preceded by a consideration of the literature in reference to the efficacy of nursing models in terms of their use within mental health nursing. A statement of philosophical beliefs is the starting point from which the model is described. The terms 'Doing-With' and 'Being-With' form the nucleus of the nurse-client relationship model that is founded upon the philosophy of Martin Buber. Buber explicated a philosophy of human relationships founded on his concepts of the 'I-Thou' and 'I-It' relationships. Two examples of the model illustrate its principles in clinical practice. The authors draw upon the work of Fawcett to argue the limitations and potential for the future development of the model described. PMID- 11933469 TI - Service user involvement in care planning: the mental health nurse's perspective. AB - A dissonance between espoused values of consumerism within mental health care and the 'reality' of clinical practice has been firmly established in the literature, not least in terms of service user involvement in care planning. In order to begin to minimize such dissonance, it is vital that mental health nurse perceptions of service user involvement in the core activity of care planning are better understood. The main findings of this qualitative study, which uses semistructured interviews, suggest that mental health nurses value the concept of user involvement but consider it to be problematic in certain circumstances. The study reveals that nurses hold similar views about the 'meaning' of patient involvement in care planning but limited resources, individual patients characteristics and limitations in nursing care are the main inhibiting factors. Factors perceived as promoting and increasing user involvement included: provision of accurate information, 'user-friendly' documentation, mechanisms for gaining service user feedback, and high staff morale. PMID- 11933470 TI - The perceptions and aspirations illicit drug users hold toward health care staff and the care they receive. AB - Over the 30 years of conflict, Northern Ireland escaped the worst excess of illegal drug trafficking and usage. However, the recent 'peace dividend' has brought with it an unprecedented rise in the availability and use of illicit drugs. With this, new problems and pressures have been brought to bear on the health service. The literature would suggest that drug users are loathed and feared by health care staff. Staff will also admit to be lacking in the knowledge and skills necessary for the delivery of appropriate support and treatment for this client group. Further, the literature has little to offer on the experiences and aspirations of drug users in relation to their treatment and the staff who care for them. In order to understand the drug users' experiences of health care and health staff, focus group methodology was employed to obtain qualitative data. A total of 20 illicit drug users from across Northern Ireland took part. Supporting the literature, all had experienced 'care' that they felt was filled with judgement, hostility and loathing. They recognized clearly the challenge they pose to health care staff. These findings indicate that there is obvious dissonance between those tasked to care for drug users and drug users themselves, with little respect being shown on either side. Results suggest that action needs to be taken to address the deficits in the knowledge, skills and values of health care professionals in relation to illicit drug users. The findings will be of interest to service providers within and outside the United Kingdom. PMID- 11933471 TI - Staff development, anxiety and relaxation techniques: a pilot study in an acute psychiatric inpatient setting. AB - Psychiatric inpatients often complain of problems with anxiety, but a computerised search of the nursing literature failed to find any publications detailing nurse-led, individual or group anxiety management work specifically aimed at psychiatric inpatient populations. In the UK, psychiatric inpatient populations are characterized by people with clinical diagnoses of schizophrenia and major affective and personality disorders. This is a very different population from that treated in the vast majority of published trials of cognitive and other approaches to anxiety management. A pilot study was conducted on four psychiatric acute admission wards to determine the practicality of treating a convenience sample of psychiatric inpatients with self-reported anxiety problems along broadly cognitive lines. Patients attended a course of three anxiety management groups (AMGs) run by nurses and were given homework and other exercises to complete. Patients reported significant reductions in anxiety after completion of the treatment. The AMGs were facilitated by staff nurses under the supervision of a clinical nurse specialist, and not by fully trained therapists as in most treatment studies relating to anxiety. Further studies, particularly randomized controlled trials, are needed to explore the efficacy and practicality of nurses delivering brief psychological interventions to psychiatric inpatients. PMID- 11933472 TI - 'Sister, I am going crazy, help me': psychodynamic-oriented care in psychotic patients in inpatient treatment. AB - A common German expression for 'crazy' means literally 'not sealed'. In psychoses, the boundaries of the ego are brittle and dissolved, the pores are open. Fragmentations occur, fusions of undefined self and object represenations are impending. The self is projected and introjected, always struggling for a sealed-up, though sufficiently permeable, space. These psychodynamics form a characteristic feature of professional psychiatric care. Providing a sealing up and cohesive relation as a basis for the return into the non-psychotic world is one of the essential tasks of psychiatric nursing. The nursing relationship forms a sort of virtual membrane which is a point of intensive and accelerated exchange. Clinical vignettes will be used to demonstrate that the creation of reliable, encouraging contact can be successful when psychotic functional patterns are understood in terms of object relations theory. PMID- 11933473 TI - An exploration of community psychiatric nursing: a Northern Ireland perspective. AB - The Audit Commission's (1994) report Finding a Place states that mental health problems are a major cause of disruption and difficulty in people's lives and that in any one year more than a quarter of all people suffer to some degree. The report asserts that the majority visit their general practitioner (GP) and less than half of the people concerned are recognized as having a mental health problem. Many recover over a period of weeks or months and are best served by community services. Only those with the most serious conditions need specialized care or admission to hospital. On a local level in Northern Ireland, the policy of reducing the number of long-stay patients began in the 1960s with a call for people with mental illness to be integrated into the community. This has had major implications for community mental health services and for community psychiatric nursing. The aim of this study into one community psychiatric nursing service (CPNS) in Northern Ireland is to inform managers and professionals about the nature and shape of this service and to facilitate wider discussion on how to plan and deliver it in the future. PMID- 11933474 TI - Education and research links between the UK and Thailand. AB - Understanding the cultural contexts of training nurses in different countries is an important element for all nurse educators. In understanding other cultures we may be better prepared to understand our own. This paper identifies some similarities and differences between nurse education in Thailand and the UK. A notable difference is that 'Thai culture' is taught in Thai educational programmes whereas equivalent programmes are perhaps missing from UK colleges of nursing. The paper closes by identifying an on-going research project into 'academic levels in nursing' that has been developed between the Royal Thai Army Nursing College, Bangkok and the School of Nursing and Midwifery Studies, University of Wales College of Medicine, Cardiff. PMID- 11933475 TI - Nursing and extrapyramidal symptoms: a critical commentary. PMID- 11933476 TI - Write! Now! PMID- 11933477 TI - Developing abilities: the future of clinical supervision? PMID- 11933478 TI - Your views count: results of the JWC reader survey. PMID- 11933479 TI - Forming a balanced opinion. PMID- 11933480 TI - Is blood pressure a clinical predictor of pressure ulcer development? PMID- 11933481 TI - Clinical trials versus laboratory studies. PMID- 11933482 TI - Ray amputation: local surgery that aims to avoid removal of the lower limb. PMID- 11933483 TI - Protecting the skin against urine and faeces. PMID- 11933484 TI - Potassium permanganate. PMID- 11933485 TI - Do leg ulcer clinics improve patients' quality of life? AB - The aim of this prospective study was to measure quality of life in patients with chronic leg ulcers. All eligible, consenting patients who attended the leg ulcer clinic at the Repatriation General Hospital (Tasmania, Australia) during the 16 month study period were included. Patients completed questionnaires on their first visit and at follow-up which occurred, on average, eight weeks later. This study found an improvement in three quality-of-life indicators--pain, sleep and mobility--over an average of eight weeks' treatment involving one to three visits to the clinic and home visits by primary care nurses. Of the 57 patients, 53 (93%) indicated that their wound had improved following attendance at the clinic. The cost-effectiveness of providing treatment in a dedicated leg ulcer clinic has been the focus of many previous studies. This study emphasises the additional benefits a dedicated clinic may have in improving quality of life in patients with chronic leg ulcers. Such benefits ought to be considered when allocating resources to leg ulcer services. PMID- 11933486 TI - Minor burn management: an Australian regional perspective. AB - Minor burns are commonly treated in hospital emergency departments. This study aimed to identify the nursing management of minor burns in hospitals in the Queensland region of Australia. A total of 200 questionnaires were distributed to hospitals with emergency departments in this region. The semi-structured questionnaire enquired about the respondents' initial and ongoing management of minor burns. Fifty completed questionnaires were returned, a response rate of 25%. Most respondents (80%) were registered nurses and the rest enrolled nurses. The respondents' practice was compared with that recommended in the literature. Their practice reflected the variations found in the literature, suggesting that some may not be as effective or efficient as others. The authors conclude that further research is required on the efficacy of products used in minor burn management. PMID- 11933487 TI - A change for the better? Measuring improvements in upgraded alternating-pressure air mattresses. AB - This study used measurements of interface pressure over time (the pressure relief index) to investigate improvements made to two alternating-pressure air mattresses. Two older models, the Nimbus 2 (Huntleigh Technology) and Pegasus Airwave (Pegasus Egerton), were compared with two new versions, the Nimbus 3 and Cairwave systems, respectively. Pressure relief was improved in seven out of 12 areas in the Nimbus 3 system, and in four out of 12 areas in the Cairwave. Significant differences in pressure relief index measurements at the heel between the Nimbus 3 and Cairwave products may explain the former's better clinical outcomes in this area, but superior pressure relief index performance at the sacrum did not predict better clinical outcomes. Therefore, different levels of pressure relief may be needed at different body sites. Individual practitioners must decide whether these improvements merit list price increases of 11% and 15%, respectively, and whether other features justify a 20% price difference between the two new systems. PMID- 11933488 TI - A comparison of hydrofibre and alginate dressings on open acute surgical wounds. AB - This study aimed to compare the performance of a hydrofibre (Aquacel) and an alginate (Sorbsan) dressing on acute surgical wounds left to heal by secondary intention. A total of 100 patients were prospectively randomised pre-operatively to receive either the hydrofibre or the alginate dressing. Dressing performance was measured at operation, at 24 hours and seven days. Parameters measured included ease of: application and removal of the first dressing; re-application on the first postoperative day; and removal and re-application one week postoperatively. The hydrofibre dressing received higher scores for all of these categories. Patients in this group also experienced less pain (mild or none) on removal of the first dressing and at one week. However, these results did not achieve statistical significance, and should be seen as a trend. Nevertheless, the authors recommend the use of hydrofibre dressings on open acute surgical wounds. PMID- 11933489 TI - Kidney transplant recipient cycles 3,000 miles for organ/blood donation. PMID- 11933491 TI - Cuts in AWP delayed until January. PMID- 11933490 TI - Tests show water quality at RCG unit below standards. PMID- 11933492 TI - Osmonics says FDA accepts action plan to correct problems at R.O. plant. PMID- 11933493 TI - Hemolytic-uremic syndrome associated with Escherichia coli O157:H7. PMID- 11933494 TI - American society of nephrology. A look at the role of medicine and industry. PMID- 11933495 TI - Personality disorders and their effect on medical treatment. PMID- 11933496 TI - How grass-roots efforts turned tobacco tax money into transplants. Social workers as advocates. PMID- 11933497 TI - New developments in hemodialysis delivery in Ontario, 1995-2000. PMID- 11933498 TI - Deciding on a transplant: not always a patient's first choice. PMID- 11933499 TI - Canadian report: organ transplants living longer. PMID- 11933500 TI - Older patients face higher risk of kidney transplant failure. PMID- 11933501 TI - D.C. area expands kidney donation opportunities. PMID- 11933503 TI - NKF and intl. Cemetery and funeral association offer organ donation info. PMID- 11933502 TI - ASTS national registry to help assess risk to liver donors. PMID- 11933504 TI - Simulect effective for pediatric transplants. PMID- 11933505 TI - Developing psychosocial care: the development and evaluation of a short course which aims to introduce psychosocial interventions to inpatient staff. AB - Following on from the author's previous article (Baker 2000) which emphasizes the need to develop psychosocial interventions within inpatient care, the author describes the process of developing and implementing a short course aiming to introduce psychosocial interventions to inpatient staff. The article also discusses the evaluation of this course and identifies some of the difficulties for acute staff in developing these skills. PMID- 11933506 TI - Substance misuse, offending and mental illness: a review. AB - The literature concerning the associations between violence, mental disorder, comorbidity and substance misuse are discussed in this review, which focuses on the findings of several international studies that demonstrate significantly higher rates of violence in substance misuse and dual diagnosis (when compared with 'single' diagnosis groups). The need for the development of an effective psychiatric nursing response in terms of assessment, liaison and joint clinical management approaches to those with a dual diagnosis is discussed in the context of United Kingdom Government legislation and policy in both forensic mental health services, and in statutory substance misuse services. PMID- 11933507 TI - Egalitarian consultation meetings: an alternative to received wisdom about clinical supervision in psychiatric nursing practice. AB - Clinical supervision (CS) has become a watchword for psychiatric nursing. Yet, there are contradictions and controversies in academic and professional discourse in relation to the nature of CS, both structure and process, its effectiveness and how this is ascertained, the preparation of supervisor and supervisee, and the quality of the supervisory relationship. The perception of such discord encouraged the authors of this paper to step outside the debate and enact a different kind of CS, which came to be known as egalitarian consultation (EC). Egalitarian consultation meetings (ECMs) were established with the postmodern turn in psychiatric nursing as a reference point. A space was created in which participants could construct their particular version of CS. The authors and six G-grade community psychiatric nurses engaged with each other for six videotaped meetings. The data from the recordings were analysed using a hermeneutic grounded theory approach (Strauss & Corbin 1994), in keeping with the style of the research, which combined the roles of researcher and practitioner for the authors. The aim was to produce local knowledge of CS. The ECMs were characterized by a sense of freedom in relation to existing rules about hierarchy and truth. The participants, each as expert in her/his own case world, produced engrossing narratives about and for practice. The group developed a cohesiveness based in closeness and this encouraged radical talk and action--a questioning of practice systems. However, for some group members, radical equated to dangerous in terms of the watchful organization and a return to 'real' work (case supervision) was observed. Innovation in relation to CS may benefit from a change in institutional culture. PMID- 11933508 TI - Peers and partners: working together to strengthen preceptorship in mental health nursing. AB - The Sussex Education Consortium commissioned a development project to identify the education and training needs of community mental health nurses (CMHNs). The project also afforded an opportunity to examine the CMHNs' perception of their role of preceptor with pre-registration diploma nursing (PRDN) students following the mental health branch. Utilizing a 'responsive evaluation' methodology, stakeholders' views were captured. As a consequence of the issues and concerns raised, the Consortium commissioned a second project, following a similar methodolgy, to explore preceptorship in greater depth. This paper reports stakeholders' views on preceptorship arrangements expressed in semi-structured interviews, focused group discussions and other fora during the two projects. Preceptor preparation and support arrangements did not meet the needs of preceptors. Many preceptors felt their role was neither valued nor acknowledged. The link lecturer role was perceived as important by preceptors; but lecturers were often unable to meet the expectations placed upon them. The volume of students and allocation arrangements caused problems for placement areas. Finding time to carry out the roles of the preceptor was a challenge for practitioners. Recommendations for improving preceptorship are discussed and developments described, illustrating how locally commissioned research may influence both education and practice in mental health nursing. PMID- 11933509 TI - Medication compliance in older individuals with depression: gaining the views of family carers. AB - Documented evidence indicates that medication non-compliance for the older person is a common and poorly understood problem. This paper reports on a pilot study, which used a focus group interview to explore carers' attitudes, experiences and perceptions regarding medication compliance for their older depressed relative. It aimed to understand the problems and challenges encountered by carers in respect of their relative being non-compliant, and provide material to develop a comprehensive educational and support package to promote antidepressant compliance. A convenience sample of seven carers participated in the focus group and a qualitative analysis of the interview identified a number of key issues. Carers had little knowledge of depression, they were not well informed regarding antidepressant medication and they took an active role to ensure compliance. The focus group highlighted the need for health care professionals to promote and encourage carers to be part of the older individuals treatment programme and provide concise and unambiguous educational information in relation to antidepressants and the importance of compliance. PMID- 11933510 TI - Community mental health nurses in Wales: self-reported stressors and coping strategies. AB - There is evidence to suggest that community mental health nurses experience stress and burnout related to their work. Previous research has been limited by a number of methodological problems. One of the problems is that studies have tended to have small or unrepresentative samples, and many researchers have only examined mental health nurses as a subset within their research, which limits the generalizability of the findings. The All-Wales Community Mental Health Nurse (CMHN) Stress Study was set up in order to address this issue. The total population of CMHNs in Wales was surveyed (N = 614) and 301 (49%) responded. The questionnaire booklet contained a number of validated instruments to measure stress, burnout, and coping, together with a demographic questionnaire. The demographic questionnaire included three open ended-questions. These questions were asked in order in determine the CMHNs' own views of the sources of stress in the workplace, and to investigate which methods they use to cope. This paper reports the findings from the content analysis of the three questions. The results from the other measures are reported in the companion paper (Edwards et al. 2000). The most frequently cited stressors included perceived workload, excessive paperwork and administration, and a broad spectrum of client-related issues. Coping strategies that CMHNs reported using included peer support, a range of personal strategies such as relaxation, and belief in self and supervision. It appears from the findings that a range of factors such as organizational pressures and factors related to working with patients are important in determining stress levels, and that informal rather than formal support networks are the preferred methods of coping. PMID- 11933511 TI - Stressors, moderators and stress outcomes: findings from the All-Wales Community Mental Health Nurse Study. AB - The All-Wales Community Mental Health Nurse Stress Study was the largest study undertaken in the UK to date to investigate stress, burnout and coping amongst the CMHN workforce. The aim of the study was to examine the variety, frequency and severity of stressors, to describe coping strategies used to reduce work based stress, and to determine stress outcomes. Questionnaires were sent out to 614 CMHNs from ten NHS Trusts throughout Wales. The response rate was 49% (n = 301). The measures used included the Maslach Human Services Survey, the CPN Stress Questionnaire, the Psychnurse Methods of Coping Questionnaire, the Rosenberg Self-Esteem Scale and the General Health Questionnaire GHQ-12. Community mental health nurses indicated that trying to maintain a good quality service in the midst of long waiting lists, poor resources, and having too many interruptions while trying to work in the office were particularly stressful items. The coping strategies that CMHNs utilized the most were having a stable home life and looking forward to going home at the end of the day, having outside interests and hobbies and talking to people that they got on well with. Forty per cent of CMHNs tended to view themselves negatively, feeling that others did not hold much respect for them. The GHQ-12 measure indicated that 35% of CMHNs had crossed a threshold of psychiatric caseness. Measured against a normative sample of mental health workers, 51% of CMHNs were experiencing high levels of long-term emotional exhaustion. Twenty-four per cent were suffering from high levels of depersonalization burnout and were not relating well to clients, whilst 14% were experiencing severe long-term feelings of lack of personal accomplishment. The results from the study provided us with a picture of stress and coping in CMHNs in Wales. Addressing these factors may help to reduce levels of experienced stress and burnout. PMID- 11933512 TI - Nurses' perceptions of multidisciplinary teamwork in acute psychiatric settings. AB - The findings of a pilot telephone survey of senior nurses' views of multidisciplinary teamwork (MDT) practice in 26 acute psychiatric admission wards within the ambit of one English health authority are reported. The survey was conducted in two stages. The first stage focused on obtaining the senior nurse's general perceptions of the organization of the work of the multidisciplinary team, and in the second stage more detailed responses to the original questions were obtained. The findings suggest that the composition and operations of the acute ward team are limited, and the relationship between the MDT and patients and their families do not entirely support the concept of increased collaboration with patients and their families. The findings are discussed within the context of the perceived crisis in acute psychiatric care in England and the ambitions for more egalitarian 'mental health' services. PMID- 11933513 TI - A pilot project to determine the demand for and utility of an out-of-hours psychiatric service run by on-call psychiatric nurses in an A&E department. AB - This paper presents the results of a 12-week pilot project that studied the demand for and utility of an out-of-hours on-call psychiatric service delivered by two experienced psychiatric nurses in an A&E department. It also considers some of the implications for a possible permanent, substantive service. An initiative to reduce the on-call hours worked by junior medical staff harked back to literature which has long suggested that health professionals, other than medical staff, can competently perform risk assessments and determine the immediate needs of patients who self-harm. The project's aims were to determine: (i) whether an on-call psychiatric nurse could satisfactorily reduce the pressures on on-call junior doctors; and (ii) the absolute demand for and nature of requested out-of-hours psychiatric assistance. Summarized details of referred patients include data on time and duration of contact, reasons for referral, outcome and other professionals involved. Of the 88 patients referred during the pilot period, 33 (37.5% of the total) were admitted to psychiatric wards and four to medical wards. Although a range of professionals was needed during the 12-week period, the nurses dealt with 42 (47.7%) of the referrals without any other professional involvement. PMID- 11933514 TI - Service users and mental health nursing. AB - A better understanding of the views of service users has become increasingly relevant in providing mental health services and for the role of the mental health nurse in meeting users' needs. The four key issues that this research endeavoured to address were: (1) What are the views and perceived needs of users of mental health services in the context of the role that users see nurses fulfilling? (2) How do student nurses perceive their future role given the changes in nurse prepartion? (3) To what extent do the answers to the first two questions converge and/or diverge? (4) Recommend actions on ways in which mental health nurse training could be improved, in light of the findings of this research. Methodologically, both qualitative and quantitative data gathering approaches were used, the former for stage one in setting the agenda and the latter for stage two by means of questionnaires. The findings indicate that the key concerns for users are issues of choice, individuality, information and quality of care in terms of positive interpersonal relationships with those who have responsibilities for helping them and who are sensitive to their real needs. Users are not asking for any costly or extravagant therapies but support that is relevant to their needs and provided in a compassionate manner. The context of their lives and the degree to which users feel integrated into society is a prominent concern. These findings have implications for how mental health nurses are educated and trained and how on qualifying they function in practice. The findings throw into question whether mental health nursing can adapt to the desired changes that are being called for. PMID- 11933516 TI - Artificial retinas implanted in blind patients. PMID- 11933515 TI - The changing of the guard in ophthalmology. PMID- 11933517 TI - Bac-stat offers a bacteriostatic ring for LASIK. PMID- 11933518 TI - Optic neuritis in children. PMID- 11933519 TI - Worksheet to calculate permanent visual impairment. PMID- 11933520 TI - Contact lens alternatives to laser vision correction. AB - As is evident from the various options discussed, patients have endless choices to correct their vision--everything from refractive surgery to high-tech contact lenses to the latest innovations in glasses. People who do not want to wear glasses can select from a wide assortment of contact lenses that provide exceptional vision, comfort, and convenience at an affordable price. Patients who never could wear contact lenses before because of having dry eyes or astigmatism or needing bifocals now can wear them because of advances in medical technology. In addition, patients who do not want to wear either glasses or contact lenses can opt for refractive surgery to correct their vision. The goal of this article is not to push patients towards any particular route but to let them know of all the choices available. Patients must weigh all of the options, including cost, convenience, and long-term stability, and then make a choice. As with anything else, nothing is without risk or price. It is up to each individual patient to weigh the risks and benefits and decide what option to choose. The best thing for a patient to do when considering any of these options is to make an appointment with his or her eye doctor and together make an informed decision about which option is the best for the patient. PMID- 11933521 TI - Lifestyle is an important factor in good ocular health. PMID- 11933522 TI - Monitoring patients at risk for drug-induced opacities. PMID- 11933523 TI - What lens is that new patient wearing? Identifying, inspecting, and verifying the parameters of rigid and soft contact lenses. PMID- 11933524 TI - [Hemangioma and extraction]. PMID- 11933525 TI - [Arachnoid bleeding and sub- and epidural hematoma]. PMID- 11933526 TI - [Lasers in dentistry 1. What is special about lasers?]. AB - Diagnosis and treatment with lasers is becoming widely spread in dentistry. This article is an introduction to a series of articles that deal with the many dental applications of lasers. The article contains a summary of presently used lasers. It also describes the pertinent features of laser radiation: parallelism of the beam and narrow line shape in the spectrum of electromagnetic radiation. Finally, it summarizes the relevant processes of interaction of laser radiation with tissue: scattering, absorption, fluorescence and frequency-doubling. PMID- 11933527 TI - [Use of the trephine for harvesting bone from the iliac crest]. AB - For closure of the cleft alveolus usually a choice between a bone graft from the chin or from the iliac crest is made. In the department of Oral and Maxillofacial Surgery of the University Hospital Groningen harvesting of bone from the iliac crest was performed with an open technique. From the results of an earlier study on the morbidity of this procedure until June 1997 it was found that in 26.1% of the cases a contour defect could be observed in the donor region. The mean width of the mature scar was 5.0 +/- 2.4 mm (range 1-10 mm). Especially these findings and the wish to further reduce postoperative pain led the University Hospital to start harvesting iliac crest bone grafts with a trephine in june 1997. In the period june 1997 through december 1999 iliac crest bone grafts were harvested in 28 cleft lip and palate patients (mean age 10.5 years; range 8-12 years) with this trephine technique. The scars were smaller in +/- 0.3 both length (mean 15 +/- 1.5 mm; range 11-24 mm) and width (mean 1.5 +/- 0.3 mm; range 1-3.5 mm). No contour defects and sensory loss occurred. Postoperative pain could adequately be relieved by local infiltration of the donor area with a 0.25% marcaine solution and analgetics. On the first day post surgery all patients could be mobilised without problems. Based on these findings and recent literature it can be concluded that the use of the trephine is the method of choice for harvesting iliac crest bone in cleft lip and palate patients. Only when large amounts of bone, a bone block or a piece of cortical bone is needed, the open technique should be used. PMID- 11933528 TI - [Diagnosis and management in case of (suspicion of) child abuse]. AB - About 5% of all children presented at a First Aid Unit in the Netherlands is a victim of child abuse. It is estimated that in the Netherlands yearly about 50.000 to 80.000 children are victims of child abuse, which is about ten children per dental practice per annum. In more than 50% of the cases there is a trauma of the head- and neck-area. Therefore, it is likely that dentists are regularly confronted with cases of child abuse. This high percentage of oro-facial trauma puts a high responsibility on the dental profession. Dentists have an ethical duty to act actively in cases of child-abuse. They may refer to an oral surgeon, consult the family physician or ask advice of the 'Advies- en Meldpunt Kindermishandeling' (Advice- and Report Centre for Child Abuse). This article describes the symptoms of child abuse and the recommended procedure for action. PMID- 11933529 TI - [Repair and revision 9. Peripheral trigeminal nerve injury]. AB - A review is given about long-term incidence of sensory disturbance in the areas of innervation of the n. trigeminus for different types of trauma and/or treatment. Diagnosis, clinical course and possible types of treatment are in addition reviewed. Regarding diagnosis, the outcome of a test on sensory function is not always related to the degree of nerve damage because methods differ in the type of afferent nerve fibers of which function is tested, and some specificity might occur in nerve damage, i.e. either thick or thin afferent fibers might be predominantly affected at a particular time. An initial quick testing of sensory function is recommended. This testing includes examining two sensory modalities, which are related to functioning of thick and thin afferent fibers respectively and which have a dichotomous yes/no outcome on the incidence of a pronounced sensory disturbance. PMID- 11933530 TI - The origins of reciprocity and social exchange in friendships. PMID- 11933532 TI - Social exchange and the developing syntax of moral orientation. PMID- 11933531 TI - Adherence to communal norms: what it means, when it occurs, and some thoughts on how it develops. PMID- 11933533 TI - Exchange and development: a dynamical, complex systems perspective. PMID- 11933534 TI - Evolutionary perspectives on the development of social exchanges. PMID- 11933535 TI - Gender and social exchange: a developmental perspective. PMID- 11933536 TI - [Biochemical markers of myocardial damage after high-energy radiofrequency ablation of atrial flutter. Value of troponin I]. AB - Creatinine phosphokinase and its MB iso-enzyme do not allow assessment of the degree of tissue necrosis after radiofrequency ablation. Cardiac Troponin I and myoglobin, new markers of myocardial lesions, are rarely used in this indication. The aim of this prospective study was to measure and compare serum markers of myocardial damage after high energy radiofrequency ablation of atrial flutter with an 8 mm distal electrode catheter. The authors measured serum cardiac Troponin I, myoglobin, creatinine phosphokinase and its MB iso-enzyme levels before and 4, 12 and 24 hours after radiofrequency ablation of common atrial flutter in 23 consecutive patients. The same markers were also measured in a control group of 9 patients undergoing electrophysiological investigation without radiofrequency ablation. All ablation procedures were simple with an average of 12.6 +/- 6 applications of radiofrequency. Bidirectional isthmic block was obtained in 22 of the 23 patients. The mean Troponin I levels were 0.01 microgram/l before ablation, 0.87 +/- 0.77 at the 4th hour (p < 0.001 versus control), 1.16 +/- 1.2 at the 12th hour (p < 0.001 versus control) and 0.7 +/- 0.63 microgram/l at the 24th hour (p < 001 versus control) after ablation. Only 13% of patients had cardiac troponin levels greater than the threshold of significant myocardial damage (> 2 micrograms/l) with a higher average number of radiofrequency applications than the rest of the group: 15.2 +/- 1 versus 11.5 +/ 5.1 (p < 0.05). An abnormally high level of markers was found in the ablation group for 19 patients (84%) with Troponin I (> 0.4 microgram/l), for 10 patients (43%) with the MB iso-enzyme (> 8 Ul/L), and for 1 patient (4%) with myoglobin (> 90 micrograms/l), and in no patient for creatinine phosphokinase (> 290 IU/L). All values were normal in the control group. The authors conclude that cardiac Troponin I is the most sensitive marker for myocardial cellular damage after high energy radiofrequency ablation of atrial flutter. The level of cardiac Troponin I seems to correlate with the number of applications of radiofrequency. PMID- 11933537 TI - [Arterial manifestations of Behcet's disease. A report of five operated cases]. AB - Vascular involvement in Behcet's disease is common, especially in deep vein thrombosis. Aneurysmal or occlusive arterial disease is, however, rare. The authors report five cases of vascular Behcet's disease reviewed over a period of 4 years (1996-1999). All these patients were men and the mean age was 38 years. The arterial disease was the presenting symptom of Behcet's disease in 3 cases. The other patients had been followed up for Behcet's disease for 4 years. The average time of onset of arterial disease was 7.2 years with respect to the presenting symptom, the range being 2 to 20 years. In all, seven arteries were involved, two patients having two arterial sites at the same time. The femoral artery was involved in 3 cases. The iliac artery was involved twice. One patient had a coronary thrombosis and the last case was of a pseudoaneurysm of the infra renal abdominal aorta which ruptured into the retroperitoneal space. Six lesions were aneurysmal. All patients underwent surgery. The histological analysis of the resected lesions showed non-specific panvasculitis. The postoperative period was marked by short and medium term complications: four prosthetic thromboses and two anastomotic pseudo-aneurysms which required several surgical procedures and which led to two deaths. These results and a review of the literature underline the need to search for Behcet's disease in all cases of aneurysmal or occlusive arterial disease in young patients, especially those born in the Mediterranean regions. Surgical treatment should not be delayed. In these inflammatory conditions with pronounced perivascular involvement, the surgery is difficult and postoperative complications are common, especially anastomotic disunion. Medical treatment has no surgical implication, but does not prevent progression of the disease. PMID- 11933538 TI - [Doppler tissue imaging in the assessment of dilated cardiomyopathy]. AB - Doppler myocardial tissue imaging is a recent technique of objective assessment of wall motion by real time measurement of intra-myocardial velocities. This technique is being evaluated in patients with dilated cardiomyopathy. Doppler myocardial tissue imaging has been used for the quantification of dobutamine stress echocardiography, for the detection of an ischaemic aetiology in patients with dilated cardiomyopathy and for non-invasive estimation of left ventricular filling pressures. At the present time, the recordings have to be analysed a posteriori and only a small number of centres have acquired expertise of these techniques at rest or during stress. Standardisation of a posteriori procedures of image processing and validation of pertinent parameters have yet to be established in this pathology. PMID- 11933539 TI - [Doppler parameters in the followup of and evaluation of patients with heart failure]. AB - Cardiac failure plays an increasing role in the daily practice of cardiologists and internists. Cardiac failure is purely diastolic in nearly 40% of cases. Whether purely diastolic or mixed, cardiac failure requires appropriate management in the long term. A step by step approach is often necessary and echocardiography with Doppler analysis of blood flow is certainly one of the most pertinent methods of following up these patients. In addition, new technologies now provide access to quantifiable regional left ventricular systolic and diastolic function: Doppler tissue and strain rate imaging will certainly be an additional help for the adaptation of strategies of management of cardiac failure in the near future. PMID- 11933540 TI - [Ectopic atrial tachycardia complicating a congenital left atrial aneurysm: value of an electro-anatomical mapping system]. AB - The authors report the case of a patient with a congenital left atrial aneurysm complicated by ectopic atrial tachycardia treated successfully by surgery. Transoesophageal echocardiography and magnetic resonance imaging provided accurate measurements of the aneurysm and its anatomical relationships. Three dimensional electro-anatomical mapping with the CARTO, system (Biosense) confirmed the shape and dimensions of the aneurysm. The system showed the electrically mute zones and the ectopic focus situated just beyond the aneurysmal neck. Surgical ablation confirmed the morphological and functional data of the imaging techniques and the patient was definitely cured. PMID- 11933541 TI - [How to reconcile limited working overtime with left main coronary stenosis?]. PMID- 11933542 TI - [Is it necessary to operate quickly in patients with significant left main coronary stenosis?]. AB - Although coronary bypass surgery is performed rapidly in the majority of cases of left main coronary stenosis to prevent cardiovascular complications, there is no reported consensus in the literature about the ideal interval between diagnostic coronary angiography and surgery. The aim of this multicenter study was to make an inventory of the serious vascular cardiovascular events which occurred between coronary angiography and surgery to determine possible predictive factors for complications and thereby identify a high risk subgroup requiring immediate revascularisation. The population comprised 283 patients with significant left main coronary disease, out of a total of 8,205 patients who underwent coronary angiography in the university hospitals of Angers, Brest, Nantes, Poitiers and Rennes. A surgical indication was retained in 216 patients. The choice of the operation date depended on clinical data in the presence of an acute coronary syndrome, patients remaining in the intensive care unit and undergoing revascularisation rapidly. Serious cardiac events (death, myocardial infarction, refractory unstable angina and left ventricular failure) occurring while waiting for surgery were rare, observed in only 6.5% of patients. Recent myocardial infarction and, to a lesser degree, unstable angina and/or left ventricular systolic dysfunction, were predictive of serious cardiac complications before surgery. The severity of the left main coronary disease and the association of right coronary disease did not increase the risk of serious cardiac events in the preoperative period. The low incidence of complications demonstrates that this strategy enables patients to wait for surgery with an acceptable risk without having to operate all patients with left main coronary disease as an emergency. PMID- 11933543 TI - [Familial hypertrophic cardiomyopathy. French study of the duration and outcome of pregnancy]. AB - Hypertrophic cardiomyopathy (HCM) is a rare, often familial condition, which may be complicated by syncope, atrial or ventricular arrhythmias and episodes of cardiac failure. This genetic disease affects young people and may be observed in women wishing for a pregnancy. The duration and outcome of such pregnancies has not been extensively studied. The authors undertook a retrospective study by questionnaire to compare the pregnancies of 41 women with HCM, a total of 150 pregnancies, with those of 39 unaffected women from the same families: a total of 132 pregnancies. None of the women died, there were no hospital admission for cardiac causes and there was no aggravation of functional status (31% of women with HCM had symptoms before pregnancy compared with 27% during pregnancy). The foetal prognosis was good with no increase in prematurity or neonatal crises. Only the women with symptoms before pregnancy had an increased risk of foetal prematurity compared with healthy women (18% versus 5%). These results indicate the good tolerance of pregnancy of women with HCM and should lead to a revision of systematic medical contra-indication of pregnancy in these patients. PMID- 11933544 TI - [Effects of preceding stable or unstable angina on hospital morbidity-mortality of myocardial infarction. Results of a continuous series of 1,910 patients]. AB - The aim of this study was to determine the effect of preceding unstable angina on the short-term prognosis of myocardial infarction based on early complications: cardiac failure, cardiac rupture, ventricular septal defect, sustained ventricular tachycardia ventricular fibrillation and hospital mortality. A continuous series of 1,910 patients admitted with 7 days of myocardial infarction was analysed retrospectively. The patients were divided into two groups according to their previous coronary history: Group A (myocardial infarction preceded by unstable angina) and Group B (myocardial infarction without preceding unstable angina). Group B was subdivided into Group B1 (myocardial infarction de novo) and Group B2 (myocardial infarction with previous stable angina). The results showed that patients with previous unstable angina (Group A) had a lower hospital mortality (7.9%) than those without (Group B) (13.3%) (p = 00017), fewer cardiac ruptures (1.1 versus 2.9%, p = 0.03) and less ventricular fibrillation (2.6 versus 4.5%, p = 0.053). Subgroups analysis showed that patients with de novo myocardial infarction (Group B1) had more sustained ventricular tachycardia than those with previous stable angina (Group B2) (5.3 versus 2.7%, p = 0.04). The authors conclude that pre-infarction unstable angina, possibly by ischaemic pre conditioning, is an independent factor of a better prognosis in myocardial infarction. PMID- 11933545 TI - [Does radiofrequency ablation of the bundle of Kent results in a lower risk of atrial fibrillation?]. AB - Radiofrequency ablation of the Bundle of Kent is a common method of treating malignant forms of the Wolff-Parkinson-White syndrome and the paroxysmal junctional tachycardia which may complicate this condition. The aim of this study was to investigate the effects of ablation of a latent or patent Bundle of Kent on the prevention of atrial fibrillation. One hundred and thirty eight patients aged 15 to 81 years of age with one or more patent (Group I) (n = 96) or latent Bundles of Kent (Group II) (n = 42) underwent successful ablation of the Bundle of Kent. Five patients in Group I (5%) and 4 in Group II (9.5%) had spontaneous paroxysmal atrial fibrillation before ablation. During electrophysiological investigation, AF was induced in 7 patients, 2 of whom had spontaneous AF in Group I and 3 in Group II. During follow-up (3 +/- 1 years), 3 patients of Group I went on to develop AF: 2 of them had never had the arrhythmia before: 4 patients of Group II, including 2 with previous AF, went on to develop AF. The risk of spontaneous AF was correlated to older ages. The authors conclude that persistence of the risk of spontaneous AF after ablation of a Bundle of Kent should be investigated especially in patients over 45 years of age. PMID- 11933546 TI - [Does radiofrequency ablation of the slow pathway prevent atrial fibrillation in cases of re-entrant intranodal junctional tachycardia?]. AB - The aim of this study was to assess the effects of ablation of the slow pathway on the eventual occurrence of atrial fibrillation (AF) in cases of intranodal junctional tachycardia (INJT). Two hundred and fifty seven patients were admitted for recurrent paroxysmal junctional tachycardia. The ages ranged from 15 to 87 years (average 54 +/- 16 years). Tachycardia was induced in all patients and the mechanism shown to be INJT in 215 patients. Twelve of these (6%) also had spontaneous paroxysmal AF. It was possible to induce INJT and AF in 23 patients during electrophysiological study (11%): of these patients, 4 had a history of AF associated with INJT. Radiofrequency ablation of the slow pathway was successfully carried out. Patients were followed up for 1 to 6 years (average 3 +/- 2 years). None were prescribed antiarrhythmic drugs. The results showed that of the 12 patients with spontaneous AF before ablation, 8 had recurrence of paroxysmal AF which required reintroduction of an antiarrhythmic treatment and a ninth patient is currently in chronic atrial fibrillation. All but one of the patients were over 65 years of age. The AF recurred 1 month to 4 years after ablation. Of the 19 patients without previous AF but with inducible AF, 2 developed spontaneous paroxysmal AF. Of the patients without previous AF and without inducible AF, 4 aged over 65 went on to develop paroxysmal AF. The authors conclude that radiofrequency ablation of the slow pathway of patients with INJT does not seem to prevent future development of AF in elderly subjects. PMID- 11933547 TI - [Guidelines for diagnosis and treatment of heart failure]. PMID- 11933548 TI - [Guidelines of the European Society of Cardiology and French Specialties concerning the diagnosis and treatment of heart failure]. PMID- 11933549 TI - [Management of heart failure by general practitioners in France. Results of the study IMPROVEMENT of heart failure (IMPROVEMENT-HF)]. AB - The IMPROVEMENT-HF programme (Improvement PROgramme on evaluation and management of Heart Failure) aimed to improve the management of patients with heart failure by general practitioners in France and in Europe. This programme had two aspects: diagnostic and therapeutic. In France, the programme had been carried out by the collaboration of 27 cardiologists and 217 general practitioners who collected the charts of 1200 patients. Among French doctors, the level of knowledge of the NYHA classification and the concept of diastolic dysfunction is poor. A majority of them request EKG and echocardiography for patients with heart failure. As expected, ACE-inhibitors are frequently prescribed, however, dosages remain low. It appears that the collaboration with cardiologists is close. In this study, some differences appear on the management between France and other countries. PMID- 11933550 TI - [CAPRICORN and COPERINUS studies]. AB - The CAPRICORN and COPERNICUS studies have confirmed the significance of betablocker treatment with carvedilol for indications which this medication did not yet possess: post-infarction with left ventricular dysfunction with or without clinical signs, and stable severe cardiac insufficiency. The reduction in morbidity and mortality obtained thanks to this therapy places it in the treatment of cardiac insufficiency in association with the angiotensin converting enzyme inhibitors which remain unsurpassed. PMID- 11933551 TI - [Evaluation of the cardiac risks in non-cardiac surgery in patients with heart failure]. AB - Cardiac insufficiency represents a major risk factor in patients about to undergo non-cardiac surgery. The post-operative mortality is linked to the severity of the pre-operative functional impairment: rising from 4% in NYHA class 1 to 67% in class IV. The operative risk is greater when the cardiac insufficiency is more disabling, the patient is older (> 70 years) and if there is a history of acute pulmonary oedema and a gallop bruit on auscultation. The use of metabolic equivalents (Duke Activity Status Index) is recommended: the functional capacity is defined as excellent if > 7 MET, moderate between 4 and 7, or poor if < 4. A non-invasive evaluation of left ventricular function is necessary in each patient with obvious congestive cardiac insufficiency or poor control under the American consensus, but it is rare that the patient has not already been seen by a cardiologist. The degree of per-operative haemodynamic constraint is linked to the surgical technique and is stratified according to the type of surgical intervention and whether or not it is performed as an emergency. An intervention duration > 5 hours is associated with an increased peri-operative risk of congestive cardiac insufficiency and non-cardiac death. Deaths from a cardiac cause are thus twice as frequent after intra-abdominal, non-cardiac thoracic or aortic surgery and the post-operative cardiac complications are six times more frequent. Numerous studies have attempted to document the impact of different anaesthetic techniques on the prognosis for the population at increased risk of post-operative cardiovascular complications. It is advisable to opt for peripheral nerve blocks. The cardiovascular morbidity and overall mortality do not differ between general anaesthetic, epidural anaesthetic or spinal nerve block. The ASA (American Society of Anesthesiologists) classification is widely used to determine the overall risk. The ASA class and the age are however too coarse as methods of evaluation for the individual risk and for giving judicious pre-operative advice. Multifactorial cardiac risk indexes such as that of Goldman allow overall evaluation (taking the patient and the intervention into account) of the peri-operative cardiovascular risk in non-cardiac surgery as a function of predictive clinical elements. Nine variables concerning the patient's history, the physical examination and several simple supplementary examinations are identified for which the relative weight is recorded under a points system. The average risk score for a given procedure is converted into an average risk for a given patient using a nomogram such as Detsky's. Surgical acts which do not impose major constraints on the cardiocirculatory apparatus (ophthalmic surgery for example) do not require supplementary examinations. The risk of post operative cardiac complications is low in the absence of the 9 risk factors defined by Goldman, as is an ischaemic syndrome (angina on light physical activity, unstable angina, myocardial infarction). Certain risk factors (jugular congestion, gallop bruit, recent myocardial infarction, non-sinus rhythm, extrasystoles, aortic stenosis) obviously require appropriate treatment beforehand. The sometimes difficult process demands a dialogue between the cardiologist and the surgeon, the recognition of the risk of surgery in a given centre, and the opinion of the patient duly informed of the terms of the discussion about him. PMID- 11933552 TI - [Prognostic value of neurohormones in heart failure in clinical practice]. AB - The decisive therapeutic advances achieved in cardiac insufficiency in recent years have been thanks to drugs affecting the different neurohormonal systems in operation. Neurohormonal activation plays a major role in cardiac insufficiency. Several neuro-endocrine mechanisms exert vasoconstrictor effects: the sympathetic system, the renin-angiotensin-aldosterone system and the endothelins. In cardiac insufficiency these effects are counterbalanced, but insufficiently, by vasodilatory agents, mainly the natiuretic peptides, EDRF (endothelium derived relaxing factor), vasodilatory prostaglandins, bradykinin and adrenomedulin. Neurohormonal activation is an excellent marker of not only the severity but also the prognosis of cardiac insufficiency. Standardisation of dosage is desirable in order to allow the use of neurohormonal drugs in a very large number of centres. PMID- 11933553 TI - [MUSTIC trial]. AB - The atrio-bi-ventricular pacing has been used for hemodynamic improvement since 1994, and the MUSTIC trial is the first controlled study assessing this concept. It collected 67 patients with severe and stable heart failure with optimised medical therapy, with sinus rhythm and without any traditional indication for pacing. After two periods of three months with blinded randomisation and cross over, the bi-ventricular pacing mode showed its superiority to the inactive mode concerning all study endpoints, for example with an improvement of the functional capacity of 23% assessed by the 6-minutes-walk test. Thus the MUSTIC trial opens up the way for morbidity and mortality studies. PMID- 11933554 TI - [Effects of bradykinin in the cardiovascular effects of angiotensin-converting enzyme inhibitors]. AB - The role of bradykinin in the cardiovascular effects of angiotensin converting enzyme inhibitors remains difficult to establish. On their haemodynamic effects, bradykinin acts during their acute administration, participating in their vasodilatation action, while during their chronic administration they act slightly or not at all. On their trophic effects, the action of the tissue kallikrein-kinin system, suggested by the results of animal experimentation, is yet to be demonstrated in man. For their effects on cardiovascular morbidity and mortality the role of bradykinin remains under discussion. Nevertheless, besides ACE inhibitors, the other therapeutic agents which increase the levels of bradykinin, such as neutral endopeptidase inhibitors, have a significant field of development in the course of cardiovascular pathologies. PMID- 11933556 TI - [Surgery in heart failure (without transplantation)]. AB - Cardiac transplant remains the treatment of reference for end-stage cardiac insufficiency. The very great disparity between the number of grafts available and the number of patients eligible to be included on the transplant list drives some of them to contemplate a surgical alternative in order to improve their clinical condition and to delay as much as possible the date of transplant. The objective is to treat surgically one or several of the lesions causing the cardiac insufficiency. It could be valvular surgery, coronary surgery or a combination of both, ventricular remodelling or cardiomyoplasty. It is coronary revascularisation surgery which gives the most spectacular results on condition that it is aimed at the myocardial zones for which viability has been demonstrated by stress echocardiography or a PET scan. PMID- 11933555 TI - [Role of angiotensin-converting enzyme inhibitors in the treatment of heart failure in the 21st century]. AB - The angiotensin converting enzyme (ACE) inhibitors have progressively stood out in a large population of heart failure patients as a gold-standard treatment, in relation with their beneficial effects on mortality and morbidity. In a recent meta-analysis published in the Lancet collecting 12,763 patients, Flather demonstrates global mortality decrease of 25% compared to placebo. This risk reduction not only concerns the mortality due to heart failure but also that due to myocardial infarction. The same goes for the morbidity. Thus, in international as well as European recommendations, ACE-inhibitors are indicated as a first lane treatment in heart failure due to systolic LV dysfunction. Nonetheless several questions remain unanswered. The ACE-inhibitors are under-utilised, not only they are under-prescribed (only 60% of heart failure patients benefit from them) but also when prescribed, the dosage (comparing to those used in clinical trials) is generally as low as the half expected. This under-utilisation seems to be related to the side effects as renal failure, hypotension or more often due to the concern of their occurrence especially in the elderly and in those with other concomitant morbidities. They are actually related in part to an under-estimation of the benefit/risk ratio. The ATLAS study suggests that high doses of ACE inhibitors are associated with a deeper reduction of morbidity without significant differences compared to low doses concerning global mortality or side effects. However this study compared very high (extreme) doses to low ones and comparison between heavy- and mid-doses remains to be performed. After the HOPE study, new indications appear promising: heart failure with preserved systolic function; patients with risk factors without heart failure: risk reduction of subsequent heart failure and reduction of mortality. Tolerance of their association with beta-blocking agents. In conclusion, the optimisation of the ACE inhibitors treatment goes through a wider prescription with higher doses, probably extended to new indications. PMID- 11933557 TI - [Heart failure at the heart of cardiology]. PMID- 11933558 TI - [Heart failure and vasopeptidase inhibitors]. AB - The morbidity and mortality of cardiac insufficiency remains such that it justifies the pursuit of finding new drugs and new sensitive techniques to slow or abolish its evolution. Bringing the vasopeptidases, such as omapatrilat, up to date results in a rational process aimed at simultaneously modulating certain interactive humoral systems. They represent drugs which simultaneously inhibit neutral endopeptidase and angiotensin converting enzyme with the effect of potentiating the natiuretic peptide system and bradykinin, and blocking the conversion of angiotensin I and angiotensin II. In the IMPRESS study, omapatrilat has been evaluated in patients with cardiac insufficiency versus lisinopril; there was no significant difference on the principal outcome measure which was exercise tolerance, however it was significantly more effective than lisinopril on the outcome measure combining death and hospital admission for deteriorating cardiac insufficiency. A wider study is underway, the OVERTURE study, which is evaluating omapatrilat versus enalapril on hospital admission and all-cause mortality. The Vanlev dossier has not yet been submitted to the regulatory authorities for obtaining its authorisation to be put on the market. PMID- 11933559 TI - [Endothelin receptor antagonists in heart failure]. AB - Endothelin 1 is a vasoconstrictor and pro-hypertrophic peptide released by the endothelium, whose secretion increases in cardiac insufficiency in proportion to the haemodynamic alteration, notably the systolic pulmonary arterial pressure. It acts by binding to 2 receptors: receptor A, responsible for the vasoconstriction effect, and receptor B, which produces vasodilatation and permits the pulmonary clearance of the molecule present in the plasma. Blocking receptor A appears logical in view of the hormonal hypothesis which prevails in the pathophysiology of cardiac insufficiency, but the significance of blocking receptor B is under discussion. The immediate effect of receptor antagonists is vasodilatation, permitting a haemodynamic improvement in patients, but the benefit on dyspnoea or the evolution of the disease has not been established. Animal models show a beneficial effect of endothelin receptor antagonists on survival, which has not been confirmed by the first studies in humans. The latter are difficult to interpret: the increase in dosage should probably to be very progressive, the optimal dosage is difficult to determine and could be lower than the doses tested. Only the result of studies underway will indicate the place of these drugs in the therapeutic arsenal for cardiac insufficiency. PMID- 11933560 TI - [Vasopressin antagonists]. AB - Vasopressin, like all the other neuro-hormonal systems, is activated in patients with cardiac insufficiency. Vasopressin attaches itself to two distinct specific receptors. It is through the intermediary of the renal V2 receptor, controlling the reabsorption of water by the collecting duct, that vasopressin finely regulates the blood osmolarity. The ubiquitous V1a receptor is essentially responsible for the vasoconstrictor effect of the hormone. Some specific antagonists for these two receptors have now been evaluated in various pathologies such as SIADH, cirrhosis or cardiac insufficiency. In this situation the mixed antagonists, anti-V1a-V2, seem more appropriate than the specific V1a or V2 receptor antagonists. The results of the first human studies are encouraging. The mixed antagonists reduce the pulmonary capillary pressure and increase diuresis and clearance of free water. But further studies are necessary to confirm these results and to demonstrate a reduction in morbidity and mortality before adding this class of medication to the therapeutic arsenal for our patients with cardiac insufficiency. PMID- 11933561 TI - [Registry of heart failure patients hospitalized in 2000. Data from the National College of Cardiology and General Hospital]. AB - The French epidemiological data on cardiac insufficiency in the hospital environment are scarce. A register collecting 1772 patients was produced by the services of the National College of General Hospital Cardiologists (C.N.C.H.G.) during two periods: autumn 1999 (November) and spring 2000 (June). It involved completing a form for each of the first 20 patients with cardiac failure hospitalized over a month. 1011 and 761 observations from 59 and 47 centres (that is 17 and 16 observations per centre) were collected during the autumn and spring periods respectively. In France, in the general hospital centres (CHG) cardiology services during the year 2000, the characteristics and the medical treatment of hospitalized patients with cardiac failure are very similar to those presented in 1998 by A. Cohen-Solal in the name of the working group "Cardiomyopathy and Cardiac Insuficiency of the French Society of Cardiology". The hospitalized patient with cardiac failure is very old, usually male, has an ischaemic cardiopathy in one in two cases, and is at stage II and III on the New York Heart Association (NYHA) scale in 83% of cases. There is practically always an electrocardiographic anomaly. Loop diuretics are prescribed nine times out of ten, digitalis one in three, anagiotensin converting enzyme inhibitors are underused being prescribed two out of three times, but an increase in the prescription of anti-aldosterone and betablockers is found. The majority of patients improve during their stay, 7.8% dying and this mortality is influenced by age, ejection fraction (FE), functional NYHA class, causal cardiopathy, and the existence of severe renal failure. The data collected by the cardiology services of the C.N.C.H.G. are representative of the profile of the population affected and are important to know in order to improve the management of these patients. PMID- 11933562 TI - [Polarographic determination of metals in firearm injuries]. AB - There was tested the use of the polarographic determination of some metals (Pb, Cu, Fe, Sb) in the skin in deceased due to multiple firearm injuries. The authors present on a real case, in which the shots through the dress of victim penetrated, the possibility to determine entry and exit wound. PMID- 11933563 TI - [Morphologic findings in chronic abuse of heroin and pervitine]. AB - In a deceased 39-year-old man with a history of 10-year consumption of heroin and pervitin the authors made histological and immunohistochemical examinations of organs focused on detection of old and recent pathological changes. In the brain they detected oedema, venostasis, inflammatory infiltrates in the wall and surroundings of some vessels and hypoxic changes of neurons with a drop or disappearance of neuron-specific enolase. The myocardium was marked by oedema of the interstitium, focal diminution to disappearance of basophilia of myocyte nuclei with increased eosinophilia of some fibres and smaller and larger foci of fibrosis formed by mature and less mature connective tissue. Immunohistochemical examination revealed focal fibrinogen deposits in myocytes. A surprise was to a certain extent the finding of dispersely distributed caspase-8 in myocytes, caspase being one of the substances signalizing apoptosis. On microscopic examination of the lungs severe haemorrhagic oedema dominated. In the liver they found acute venostasis and chronic inflammatory changes with connective tissue proliferation in the portal areas. The finding in the kidneys suggested acute tubular nephrosis. The authors discuss the problem of direct toxicity of the ingested drugs, the influence of repeated states of hypoxia and infection. PMID- 11933564 TI - [Repair and revision 8. Relapse of lower incisors: retreatment?]. AB - Research into the long-term stability of orthodontic treatment at the University of Nijmegen (the Netherlands) until 10 years after retention has shown that nearly 50% of the total relapse takes place the first two years after retention. After that period certain stability is reached except for the lower front teeth. Ten years after the retention phase their position is even worse than at the start of treatment. The changes in lower front teeth alignment are the result of relapse, but also of normal physiological changes during ageing of the dentition. Therefore it is questionable whether late mandibular incisor irregularity should be (re)treated. Relapse of mandibular incisor alignment shortly after debonding can be restored by tightly tying the rotated incisor(s) to the C-C bar with a steel ligature, by using a spring-retainer or by rebonding of brackets to the lower front teeth. After correction of the incisor position the C-C bar should be bonded to all lower front teeth. (Re)treatment of mandibular incisor irregularity at an older age asks for more complicated treatment mechanics. PMID- 11933565 TI - [Clinical underestimation of the prevalence of approximal dentin lesions and inadequate restorations]. AB - The aim of this study was to determine the additional diagnostic value of bitewing radiographs compared to clinical examination, with respect to the presence of dentine lesions and inadequate restorations for different age groups. Subjects were 14-, 17-, 20-, 23-years-old, or were in the age groups of 25-34 and 35-54 years, who participated in 2 clinical epidemiological surveys. After obtaining informed consent, bitewing radiographs were made of approximately 25% of the participants (n = 663). The extra diagnostic yield of the bitewing radiographs varied between a factor 1.6 and 7.2 for approximal dentine caries diagnosis of untreated surfaces, and between 2.3 and 5.9 for inadequate restorations. In conclusion, a considerable increase in the numbers of approximal dentinal lesions and inadequate restorations for all age groups was found. PMID- 11933566 TI - [Recognition of malignant skin tumors of the head and neck]. AB - Skin cancer is the most common malignancy in man. The sun-exposed surfaces of the head and neck are the areas most frequently involved. Since skin cancers in most instances are easily recognized, they may be diagnosed at an early stage. The dentist may play a role in the early detection of skin cancers of the head and neck. PMID- 11933567 TI - [NVT/NiVVT Fall Congress. "The child in the chair"]. PMID- 11933568 TI - [A mysterious swelling on the upper lip]. PMID- 11933569 TI - [Cerebrovascular accident]. PMID- 11933570 TI - Blood film preparation and staining procedures. AB - The blood film is one of the world's most widely and frequently used tests and has undergone remarkably few changes since its introduction in the late 1800s. This article gives direction and some standardization in the preparation of blood films used for morphologic evaluation in the clinical laboratory. Methodologies that can be labeled as being state-of-the-art and methods applicable for laboratories in developing countries are quoted. PMID- 11933571 TI - Clinical utility of the band count. AB - Enumeration of band neutrophils has a long clinical tradition as a diagnostic test for bacterial infection. Yet, the band count is a nonspecific, inaccurate, and imprecise laboratory test. Review of the literature provides little support for the clinical utility of the band count in patients greater than 3 months of age. The white blood cell count and the automated absolute neutrophil count are better diagnostic tests for adults and most children. Absolute numbers of bands are required for the Rochester criteria, a diagnostic algorithm for acutely ill, febrile children less than 3 months of age. No studies, however, assess the independent contribution of bands to the performance of the algorithm, or the use of the automated total neutrophil count as a replacement for the band count. Band counts also are required to calculate an immature to total neutrophil ratio (I:T ratio), an index widely used to aid in the diagnosis of neonatal sepsis. Studies, however, show a wide range of sensitivity and specificity for the I:T ratio, indicating variable performance. In the near future, rapid analysis of inflammatory factors, adhesion molecules, cytokines, neutrophil surface antigens, or even bacterial DNA may be superior alternative tests for the early diagnosis of sepsis. PMID- 11933572 TI - Peripheral blood findings in chronic myeloproliferative disorders. AB - Chronic myeloproliferative disorders are now frequently detected on routine total blood count. A careful analysis of PB smears allows the diagnosis in many cases, and helps to classify most of them. The peripheral blood parameters may be of prognostic value, especially in AMM. During the follow-up of CMPD, modifications of the PB are of great interest to detect an evolution toward a myelofibrosis or a blastic phase. PMID- 11933573 TI - The wright-giemsa stain. Secrets revealed. AB - The colorful story of the development of the Wright-Giemsa stain is retold. Dramas are replayed, secrets are exposed, and laurels are properly returned to scientists long forgotten. The delicately balanced chemical composition of the stain, once enigmatic, is defined. Finally, an attempt is made to unshroud some of the mystery surrounding the staining method; tried-and-true procedures and useful snippets of laboratory lore are provided. Scientific explanations and reliable methods aside, however, the stain continues to possess a certain mystique, seemingly consisting of equal parts technical expertise, art, and magic. PMID- 11933574 TI - Acute leukemia. AB - This article provides a review of the acute leukemias with updated basic and practical information. The main emphasis is on techniques used to arrive at the correct diagnosis. Although morphology and cytochemistry were the mainstays of diagnosis in the past, new developments in immunophenotyping, cytogenetics, molecular biology, and in vitro assays have improved the understanding of this disease dramatically and enable the identification of new entities with distinct clinicobiologic features. PMID- 11933575 TI - Assessment of platelet numbers and morphology in the peripheral blood smear. AB - Automated systems are currently in widespread use for the assessment of patients' complete blood counts. The evaluation of the peripheral blood smear, however, still constitutes a pivotal tool in the evaluation of patients with hematologic disorders. This article focuses on disorders affecting the number or morphology of platelets as assessed by evaluation of a peripheral blood smear and also outlines some of their important clinical findings. PMID- 11933576 TI - Peripheral blood manifestations of lymphoma and solid tumors. AB - Although the peripheral blood film has limited use in this age of technologic testing, most new tests are ancillary and cannot be interpreted accurately without examining the peripheral blood film. The peripheral smear plays a critical role in the diagnosis and management of many lymphoproliferative disorders, although playing more of a general function in nonlymphoid malignancies to evaluate cytopenias. PMID- 11933577 TI - Red cell morphology and the peripheral blood film. AB - Red cell morphology as determined by microscopic examination of a stained blood film is an old and traditional approach to the evaluation of an anemic patient. The examination of a well-made and well-stained peripheral blood film remains an important and vital tool in the evaluation of the anemic patient and provides direction for the subsequent laboratory evaluation of the patient's anemia. This article provides a review of the important red cell changes necessary for evaluation in determining the cause of anemia. PMID- 11933578 TI - Infectious diseases manifested in the peripheral blood. AB - Although primary diagnosis of infectious disease is uncommonly made from morphologic examination of a blood smear in the United States, knowledge of the distinctive morphologic features of various organisms, coupled with an understanding of the clinical and epidemiologic features of various disorders, permits recognition and diagnosis of uncommonly encountered infections. Furthermore, nonspecific manifestations of infection may provide an important clue in guiding a further diagnostic work-up. PMID- 11933579 TI - Peripheral blood film review. The demise of the eyecount leukocyte differential. AB - The automated hematology analyzer with CBC and differential results has replaced the traditional manual or individual assay methods for hematologic parameters and the eyecount leukocyte differential as the initial screening and detection system for hematologic abnormalities in modern hospitals and clinics. The traditional review of all automated hematology instrument results by preparation, staining, and microscopic examination of a blood film has disappeared in most institutions. The reasons are the more accurate detection of specimens with distributional or morphologic abnormalities by the instruments than by the traditional eyecount method. The opportunity for a clinician to request a microscopic examination of a blood film, whether or not it is flagged, must be preserved, because the clinician's knowledge of the patient's history, physical findings, and current or prior therapy may indicate review to discover an abnormality that may not have been apparent from the instrument results alone. There has also been a dramatic reduction of the numbers of medical technologists and technicians in medical laboratories. Automation of the CBC and differential counts has reduced the number of technologists needed for performance of these tests. But other factors have had a negative effect, such as the necessity to reduce costs. Consolidation of hematology and chemistry laboratories in core laboratories may produce savings in labor costs, but may also create problems of creating and maintaining areas of expertise, such as hematologic morphology, because of the cross-training required and the necessity of personnel to do all things. This article suggests and documents a number of measures that can be infinity stituted by the laboratory and by clinicians to reduce the number of eyecount differentials and blood film reviews that need to be performed. The first effort is to convince clinicians that valid data exist that confirm that a policy of allowing the laboratory to initiate blood film review based on findings of the CBC and automated differential is a more sensitive and accurate method of detecting patients with blood film abnormalities than routine blood film review of all specimens by technologists. Clinicians need to recognize that daily differential results or differentials at intervals of less than a week are not medically necessary in most patients. The laboratory, however, must provide opportunities for the clinician to request differentials at any time for specific medical reasons. The laboratory must establish the validity of screening criteria for detection of distribution and morphologic abnormalities of leukocytes by clinical correlation studies or adopt criteria established by laboratories with the same instrumentation and which have conducted clinical evaluations. A final observation on the eyecount differential is that it was the only way to identify cell types and their relative proportion for nearly 100 years. Cells were identified by their shape, intracellular structures, and staining characteristics. Many studies were able eventually to correlate some aspect of each cell type's function with their morphologic appearance. It has also been learned that the bone marrow is the source of production of most circulating cells and a great deal of the controls of cell production and release into the peripheral blood have been learned. But leukocytes have many functions, almost none of which are performed in the peripheral blood. The peripheral blood is mainly a conduit from the bone marrow to the tissues where the leukocytes perform their function in the case of the neutrophils and monocytes. It is mainly a recirculation and redistribution system for lymphocytes that usually receive their instructions from antigen processing cells in the tissues and allow these modified cells to home to sites where their functions occur. Cellular morphology and staining characteristics tell little about the maturation stage and functional capabilities of leukocytes. One cannot tell the difference between a band and a segmented neutrophil or whether a lymphocyte is a T or B cell on the conventional eyecount differential. One cannot tell the mature granulocyte of a patient with chronic myeloid leukemia from a normal mature neutrophil. Increasingly, techniques are being developed to identify better the maturation stages of cells and association with specific functional capabilities by flow cytometric techniques. The neoplastic nature of some normal-appearing leukocytes can be identified by techniques, such as fluorescent in situ hybridization. With the rapid advances in many approachs to understand the nature and functional capability of leukocytes, the eyecount differential with the traditional Romanowsky stain may be past the apogee of its ascent and beginning its trip into history along with the hemocytometer counting chamber and the Sahli pipet. The development and implementation of new laboratory cornerstone techniques for diagnosis of hematologic disease are eagerly awaited. On the other hand, the red cells and platelets exist to function in the peripheral blood. More emphasis is needed in the development of automated methods of determining the nature and functional capabilities of these true blood cells as part of the CBC. PMID- 11933580 TI - Automated image processing. Past, present, and future of blood cell morphology identification. AB - Automated image processing analysis for leukocyte differential counting started 30 years ago principally as a mimic of the traditional microscopic method. Several types of systems were used in the 1970s and 1980s. In the late 1990s, two new image processing systems were developed with new technology for cell image analysis. They possess an intelligent neural network software and can be connected to an Ethernet for telehematologic diagnosis and consultation. PMID- 11933581 TI - The virtual blood film. AB - The computer and the digital camera offer unprecedented possibilities for improving hematology education, research, and patient service. Peripheral blood smear images of exceptional quality can be acquired rapidly and conveniently from the peripheral blood smear with a modern, high-resolution digital camera and a quality microscope. Digital cameras use CCD or CMOS image sensors to measure light energy and additional circuitry to convert the measured information into a digital signal. Because digital cameras do not use photographic film, images are immediately available for incorporation into web sites or digital publications, printing, transfer to other individuals by e-mail, or other applications. Several excellent consumer digital still cameras are now available for less than $1000 that capture high-quality images comprised of more than three megapixels. These images are essentially indistinguishable from conventional film images when viewed on a quality color monitor or printed on a quality color or black and white printer at sizes up to 8 x 10 in. Several recent dedicated digital photomicroscopy cameras provide an ultrahigh quality image output of more than 12 megapixels and have low noise circuit designs permitting the direct capture of darkfield and fluorescence images. There are many applications of digital images of peripheral blood smears. Because hematology is a visual science, the inclusion of quality digital images into lectures, teaching handouts, and electronic documents is essential. A few institutions have gone beyond the basic application of digital images to develop large electronic hematology atlases; animated, audio enhanced learning experiences; multidisciplinary Internet conferences; and other innovative applications. Digital images of single microscopic fields (single frame images) are the most widely used in hematology education at this time, but single images of many adjacent microscopic fields can be stitched together to prepare zoomable panoramas that encompass a large part of a microscope slide and closely stimulate observation through a real microscope. With further advances in computer speed and Internet streaming technology, the virtual microscope could easily replace the real microscope in pathology education. Interactive, immersive computer experiences may completely revolutionize hematology education and make the conventional lecture and laboratory format obsolete later in this decade. Patient care is enhanced by the transmission of digital images to other individuals for consultation and education, and by the inclusion of these images in patient care documents. In research laboratories, digital cameras are widely used to document experimental results and obtain experimental data. PMID- 11933582 TI - Reticulocytes. Their usefulness and measurement in peripheral blood. AB - The eyecount reticulocyte count has been replaced by semi-automated and fully automated methods in laboratories of nearly all developed nations, except for small-volume laboratories and office laboratories in which they are not as cost effective. The eyecount method remains the reference method as defined in NCCLS document H44-A. PMID- 11933583 TI - Artifacts that may be present on a blood film. AB - Many artifacts or pseudoartifacts may be noted during the examination of a blood film. Artifactual results also may be generated by automated hematology analyzers, which in turn may be investigated by blood film examination. Some artifacts are misdiagnosed, and this in turn leads to inappropriate investigations or treatment. An awareness of the spectrum of spurious or artifactual results may help to prevent such an unfortunate occurrence. PMID- 11933584 TI - Hepatic imaging. An overview. AB - For optimal detection and characterization of focal or diffuse liver disease, it is essential to obtain the most appropriate imaging test in the correct clinical setting. Access to clinical information and medical history is, therefore, essential. Moreover, familiarity with currently available modalities for imaging the liver [figure: see text] allows the optimal use of the technical advances in ultrasound imaging, CT scanning, MR imaging, and nuclear scintigraphy technology and contributes to improved diagnostic accuracy. PMID- 11933585 TI - Benign liver neoplasms. AB - A variety of benign focal liver lesions are easily characterized with currently available imaging techniques and contrast agents. The most common benign liver lesions, such as hemangioma, bile duct cyst, and FNH, reveal characteristic cross sectional imaging features that allow an accurate diagnosis. For atypical variants and more uncommon lesions, including HCA, angiomyelioma, infantile hemagioendothelioma, and mesenchymal hamartoma, integration of clinical data can often help in the interpretation of imaging studies. Finally, for the remaining lesions, such as hepatic adenomatosis, the imaging findings may not be specific enough to negate the need for a tissue biopsy. PMID- 11933586 TI - Malignant liver tumors. AB - The primary hepatic malignancies are a diverse group of neoplasms with distinctive clinical and pathologic features. Imaging of the primary hepatic malignancies continues to be challenging. Ultrasonography, CT scanning, and MR imaging play complementary roles in the evaluation of these patients. Many [figure: see text] of these neoplasms have distinctive imaging features that may permit diagnosis. In most instances, however, biopsy is required for definitive diagnosis and treatment planning. PMID- 11933587 TI - Computed tomography and magnetic resonance imaging of hepatic metastases. AB - The detection and characterization of liver metastases is well performed with either computed tomography or magnetic resonance imaging. The administration of intravenous contrast is essential for almost all indications, with multiphasic imaging aiding in lesion characterization and detection. The use of multidetected CT (MDCT) provides the ability for optimized vascular and multiplanar imaging, but has also resulted in increased examination complexity. Tissue-specific MR contrast agents can yield the highest rate of lesion detection and thus may be useful in presurgical planning. PMID- 11933588 TI - Ultrasonography of the liver. An update on new applications. AB - Sonography is highly useful in evaluating the patient with severe liver disease before and after the placement of a TIPS and before and after organ transplantation. Efforts to use ultrasound imaging to evaluate for primary and metastatic lesions to the liver have been reviewed. With its Doppler and color flow capabilities, ultrasound imaging will remain an important modality for hepatic imaging, especially in the evaluation of portal vein patency and hepatic artery thrombosis. PMID- 11933589 TI - Diffuse liver disease. AB - During the last decade, the role of the radiologist in evaluating patients with diffuse liver disease has increasingly expanded. In many cases, the management choices for the hepatologist in the imaging work-up of a patient with suspicion of a diffuse liver disease have significantly widened. In some instances, imaging may point directly to the diagnosis; in many instances, imaging helps narrow the differential diagnosis or is crucial in the follow-up of patients. Although some rare entities still have nonspecific radiologic features, the imaging pattern, in combination with appropriate clinical information, may provide the most likely diagnosis. PMID- 11933591 TI - Unique imaging issues in pediatric liver disease. AB - In many clinical scenarios, liver imaging does not differ as greatly in children as in adults. Common indications for liver imaging in children include trauma, suspected mass, pre-transplantation studies, monitoring after liver transplantation, jaundice, or liver dysfunction. This article highlights areas in which pathology or imaging approach in children differs from that seen in adults. Topics covered include imaging of a suspected hepatic mass, neonatal jaundice, and segmental liver transplantation. PMID- 11933590 TI - Inflammatory disease of the liver. AB - Imaging and imaging guided intervention have revolutionized the management of hepatic inflammatory diseases. Pyogenic abscess is preferentially treated percutaneously. Radiologic techniques are crucial for the diagnosis of amebic liver abscess and infectious conditions of the liver in immunocompromised patients. PMID- 11933592 TI - Imaging of hepatic transplantation. AB - Hepatic transplantation has become the treatment of choice for advanced irreversible liver disease. More than 4,000 hepatic transplantations were performed in the United States in 1997 and more than 11,000 are awaiting transplantation. Graft endurance and overall patient survival has been steadily improving, and between 1992 and 1994, 82% of the patients who received a liver transplant survived for at least a year. Today, liver transplant patients have a five-year survival rate of approximately 75%. The improvement in survival can be attributed to better patient selection and preparation, advances in organ preservation, improved immunosuppressive therapy agents and refinement of surgical techniques. In this article, we will address the hepatic parenchyma and vascular structures that should be evaluated prior to and following liver transplantation, the range of expected anomalies and abnormalities, and the utility of each of the three main imaging modalities, namely ultrasonography, computed tomography and magnetic resonance imaging in this assessment. PMID- 11933593 TI - Liver imaging. A surgeon's perspective. AB - Advances in the diagnosis and treatment of liver lesions have improved therapy for a broad range of clinical conditions, many of which could not be effectively treated in the recent past. These advances are the result of better surgical techniques as well as diagnostic imaging. This article discusses the anatomy of the liver and the clinical evaluation of patients with liver lesions. Common benign and malignant liver lesions are presented with radiologic characteristics and treatment options. PMID- 11933594 TI - Current techniques of computed tomography. Helical CT, multidetector CT, and 3D reconstruction. AB - The many recent advances in CT technology have secured its position as the modality of choice in routine liver imaging and have improved its performance in several problem-solving applications. In addition, improvements in postprocessing software (e.g., in speed, efficiency, and automated algorithms) have increased their use in clinical practice. Multiplanar reformations, 3D renderings, and high quality CT angiographic displays have become extremely valuable both in image interpretation and in communicating information to surgeons and referring physicians. PMID- 11933595 TI - Liver imaging. A hepatologist's perspective. AB - Numerous diagnostic modalities may be employed in the assessment of liver disease. In this article we outline radiologic approaches to several clinical problems including the evaluation of abnormal liver function tests and jaundice, evaluation of liver masses, and management of the patient with cirrhosis and portal hypertension. PMID- 11933596 TI - Magnetic resonance imaging techniques. AB - In this article we describe state-of-the art techniques for magnetic resonance imaging of the liver. T1-weighted, T2-weighted, and heavily T2-weighted pulse sequences are discussed. Gadolinium-enhanced hepatic parenchymal imaging and magnetic resonance angiography are also described. A comprehensive MR imaging examination of the liver affords evaluation of focal and diffuse hepatic parenchymal disease, biliary disease, and vascular pathology. PMID- 11933597 TI - Magnetic resonance imaging. Liver-specific contrast agents. AB - MR imaging with new liver-specific contrast agents will probably be the imaging modality used in the future to detect focal liver lesions. The detection of HCC will probably be improved by using specific hepatobiliary agents, but the exact technique remains to be determined. New liver-specific contrast can differentiate some benign lesions from malignant ones and can assist in making a final diagnosis. In certain circumstances, liver-specific contrast agents can be used to evaluate hepatic vessels, the biliary tract, and hepatic function. New applications are also expected. PMID- 11933598 TI - Interventional radiology procedures in the liver. Biopsy, drainage, and ablation. AB - Radiologically guided interventional procedures in the liver have continually increased, in number and variety, over the years. Factors promoting these advances include new technology, innovative ideas, and growing acceptance by clinicians and patients. Percutaneous biopsy and drainage procedures are firmly established techniques with low complication rates. Ablation by injected substances is useful for treating certain tumor types. The most exciting development is the introduction of percutaneous thermal techniques for tumor ablation. Although more experience is needed to optimize the use of focal thermal ablation, early results seem promising. PMID- 11933600 TI - Development of a capitated system for reimbursing physicians under a professional risk HMO contract. PMID- 11933599 TI - Racial/ethnic diversity management and cultural competency: the case of Pennsylvania hospitals. AB - Major demographic trends are changing the face of America's labor pool, and healthcare managers increasingly face a scarcer and more diverse workforce. As a result, healthcare organizations (HCOs) must develop policies and practices aimed at recruiting, retaining, and managing a diverse workforce and must meet the demands of a more diverse patient population by providing culturally appropriate care and improving access to care for racial/ethnic minorities. Ultimately, the goal of managing diversity is to enhance workforce and customer satisfaction, to improve communication among members of the workforce, and to further improve organizational performance. Research on diversity management practices in HCOs is scarce, providing few guidelines for practitioners. This study attempted to close that gap. Results show that hospitals in Pennsylvania have been relatively inactive with employing diversity management practices, and equal employment requirements are the main driver of diversity management policy. The number and scope of diversity management practices used were not influenced by organizational or market characteristics. The results suggest that hospitals need to adopt diversity management practices for their workforces and need to pay particular attention to marketing and service planning activities that meet the needs of a diverse patient population. PMID- 11933601 TI - Interview with Judy Pelham, FACHE, president and CEO, Trinity Health, Novi, Michigan. interview by Kyle L. Grazier. PMID- 11933602 TI - Providing high-quality healthcare: are we up to the task? PMID- 11933603 TI - A changing workforce calls for twenty-first century strategies. PMID- 11933604 TI - Ethical dilemmas in home care case management. AB - The role of case manager is fraught with challenges in a healthcare environment characterized by rapid aging of the population, a move against institutionalization of seniors, and the need to contain healthcare costs. This study examined experiences of 89 case managers through focus groups in five urban and five rural regions of Canada to identify ethical dilemmas and issues encountered in their role. Overall, the case managers expressed frustration for the lack of support for their work as evidenced by inadequate resources and few agency policies. The analysis of the focus group data revealed four main themes in relation to ethical concerns and dilemmas: (1) issues related to equity, (2) beneficence, (3) non-maleficence, and (4) autonomy and power imbalances. The situation facing these workers is grave and steps must be taken to provide them with ongoing training, support, and resources to continue in this vital role. System changes that would reduce some of the ethical conflicts experienced by case managers include funding for long-term care to keep pace with growing demands, better management of client waitlists to ensure that the most needy are given the highest priority, more supportive housing options that provide for some on-site coordination of services, better opportunities for health promotion, and better interdisciplinary teamwork so that case managers are not left making decisions in the absence of other key service providers. PMID- 11933605 TI - Rekindling the flame: routine practices that promote hospital community leadership. AB - Because of the need of focus on their financial survival, hospitals and health systems are not the popular community-oriented institutions they once were--too many have set aside their mission to promote and protect the health of their communities. By conducting on-site interviews with CEOs, other executives, board members, and community partners in seven hospitals across the nation, we discovered 25 management and governance practices used by hospitals that excel in relating to their communities. We have grouped these practices for promoting community health into six functions: (1) identifying community needs; (2) setting goals; (3) allocating financial and human resources; (4) educating leaders, staff, and community members; (5) measuring outcomes; and (6) sustaining the mission. Specific examples are given that describe what we learned. Ultimately, we hope to engender a dialog about other management and governance practices that have proven successful in promoting hospitals' involvement in community health. PMID- 11933606 TI - [Dentists and peer review: results of a descriptive study on perceived effects of peer review]. AB - In the Netherlands over 20% of dentists participate in peer review groups. The peer review method is highly structured and consists of: setting standards for good dental care, measuring and evaluating performances against the standards and making improvements if necessary. An independent research institute (Nivel) investigated the perceived effects and advantages of and barriers to peer review. A postal questionnaire was sent tot 278 dentists who participate in peer review (response 60%). The results showed many effects of peer review. The most frequently reported effects were: more insight into the performances of colleagues, increased inter-professional contacts, a more critical attitude to performances, increased professional expertise and new inspiration and motivation. The respondents reported improvements in practice-organisation as well as in technical aspects of dental care. However, many dentists also mentioned some barriers to peer review. The method was evaluated as rather complex and it appeared to be difficult to reach consensus about standards for good dental care. In general, respondents perceived a positive balance between advantages and disadvantages of peer review, however many dentists were dissatisfied with the lack of financial compensation for participation in peer review. PMID- 11933607 TI - [Dentists and mutual practice visitation. Results of a descriptive study on the effects on mutual visitation by dentists]. AB - In 1998 mutual practice visitation has been introduced in the Dutch dental practice as a method to improve quality of dental care. The method is called 'mutual visitation' because groups of three dentists visit and evaluate each others practices. An independent research institute (Nivel) investigated the perceived effects, advantages and barriers of visitation. A postal questionnaire was sent to all 235 dentists who, by that time, participated in visitation. The response was 57%. The results showed that the effects reported most frequently were: a more critical attitude to dental practice, more insight into practices of colleagues and actual improvements in dental care. The majority of the respondents considered visitation suitable to improve practice-organization. A minority considered visitation suitable to increase professional expertise. The mutuality of the visitation was highly valued by the participants. The main disadvantage of visitation appeared to be the time investment (a mean of three days per dentist). In general, the respondents held the view that the positive effects outweighed the time and efforts, however, many dentists regretted the lack of financial compensation for visitation. PMID- 11933608 TI - [Early symptoms of Burkitt's lymphoma]. PMID- 11933609 TI - [Diabetes mellitus 2]. PMID- 11933610 TI - [Diagnosis of occlusal caries lesions using laser fluorescence measurements]. AB - Recently, a device for detecting occlusal caries lesions (DIAGNOdent) has been introduced. The reproducibility and validity of this laser-fluorescence device were investigated. In the in vivo part of the study, 45 sites at the occlusal surfaces of permanent molar teeth in 13 patients were measured by 2 observers using 2 DIAGNOdent devices, one produced in 1998 and one in 1999. The interobserver reliability between both devices and both observers was established. In the in vitro part of the study, 49 permanent molars were measured by 2 observers using 2 DIAGNOdent devices. In addition, visual inspection was performed. The teeth were sectioned to measure the histological depth and area of the caries lesions. The reproducibility of both DIAGNOdent devices was high, and so was the interobserver reliability. The correlation between DIAGNOdent measurements and the actual depth of the caries lesions was lower than that of visual inspection. The correlation with the enamel part of the lesion exceeded that of the dentine part. It was concluded that the validity of the DIAGNOdent, expressed as the area under the Receiver Operating Characteristic (ROC) curve, was not statistically significantly different from that of visual inspection. Because of the high reproducibility, dental practitioners who wish to use the DIAGNOdent for monitoring caries lesions, this investigation indicates that an old device may be replaced by a new one, provided that the same tip will be used. PMID- 11933611 TI - [Onlay grafts in combination with endosseous implants in severe mandibular atrophy. A prospective radiological study]. AB - The placement of endosseous implants in combination with iliac crest onlay grafting of the anterior mandible is one of the treatment modalities for extreme atrophy of the mandible. The remodelling of these onlay grafts was studied using standardized extraoral oblique lateral cephalometric radiographs (OLCRs). A group of 8 patients was used in this prospective study. The measurements obtained from the OLCRs indicated the existence of the following stages in the process of remodelling of the bone grafts: 1. decrease in thickness and radiographic density of the (upper) cortex of the graft, predominantly during the first half year after grafting; 2. no significant changes in the radiographic density of the upper spongeous part of the graft; 3. a mean decrease of approximately 25% in the overall thickness of the graft, particularly during the first half year; and 4. an increase in the radiographic density of the lower part of the spongeous bone in the second half year after grafting. It is concluded that the remodelling of the graft has a predictable pattern in time. Densitometric measurement using standardized OLCRs can be a useful tool to evaluate quantitative changes of bone grafts to the mandible. The treatment described can be used on very strict indications only. PMID- 11933612 TI - Polishing your image. PMID- 11933613 TI - Five steps to digital safety. PMID- 11933614 TI - Converting mg/m3 to ppm and vice versa. PMID- 11933615 TI - Breaking the hearing protection barrier. PMID- 11933616 TI - Avoiding overattenuation. PMID- 11933617 TI - Adhesive safety whys and hows. PMID- 11933618 TI - In top form. Advances in head protection raise comfort, safety, and style for workers. PMID- 11933619 TI - Flammable liquid safety. PMID- 11933620 TI - Ripe for a challenge. Does OSHA's lockout/tagout standard really require equipment-specific energy control procedures? PMID- 11933621 TI - Leverage your LOTO logic. PMID- 11933622 TI - Another reason to keep hard hats off the rear deck. PMID- 11933623 TI - A new approach for beating the heat. PMID- 11933624 TI - Safety and the professional welder. PMID- 11933625 TI - Procter & Gamble's transformation. PMID- 11933626 TI - Reliable barrier. PMID- 11933627 TI - [Should all groin hernias be operatively treated?]. PMID- 11933628 TI - [Laparoscopic hepatectomy]. AB - AIM: To report the current indications and techniques of laparoscopic liver resections, and assess the results of this technique by reviewing international literature. REVIEW OF THE LITERATURE: About 200 laparoscopic hepatectomies have been reported from 1991 to 2001. 102 resections were performed for malignant tumours, and 84 for benign tumours. Global conversion rate was 7% (13/186). Morbidity rate was 16.1% with two cases of possible gas embolisms (1.1%). Mortality rate was 0.54% (1/186 patients). Mean hospital stay was 7.7 days. CONCLUSION: Laparoscopic hepatectomy is feasible, with a morbidity and mortality rate comparable to open procedures according to a careful selection of patients. However, prospective randomized trials are still needed to confirm those results, especially for resection of metastasis or malignant tumors. Evolution of laparoscopic hepatectomies will probably depend on the development of new techniques and instrumentations. PMID- 11933629 TI - [Laparoscopic hepatic resection. Useful or futile?]. PMID- 11933630 TI - [Hiatus calibration decreases postoperative dysphagia after laparoscopic fundoplication: case-report study]. AB - STUDY AIM: The risk of dysphagia after antireflux surgery seems to be increased with laparoscopy compared with open surgery. Calibration of the hiatus is usually done by the surgeon's finger during open surgery. The aim of this study was to assess the results of laparoscopic calibration with a Fogarty balloon catheter. PATIENTS AND METHODS: Between 1999 and 2001, 21 patients had a laparoscopic Toupet 240 degrees fundoplication with hiatus calibration using a 4 ml-inflated 8G Fogarty balloon catheter. These patients were compared with a group of 21 patients without hiatus calibration, matched for age, sex, preoperative dysphagia and esophageal dysmotility. Judgment criteria was early and/or late postoperative dysphagia (> 3 months). RESULTS: Median follow-up was 13 months. The rate of early dysphagia with and without calibration were 66% and 48% respectively (NS). Median duration of early dysphagia with and without calibration were 25 and 43 days respectively (p = 0.05). No patient with calibration had late dysphagia. One patient (5%) without calibration had unexplained late dysphagia for 2 years. He had preoperative esophageal dysmotility without oesophagitis. CONCLUSION: Hiatus calibration with a Fogarty balloon catheter decreased early postoperative dysphagia duration after Toupet laparoscopic fundoplication. This easily reproducible technical point standardizes the hiatus closure and should be recommended. PMID- 11933631 TI - [Laparoscopic fundoplication for gastroesophageal reflux disease: retrospective study of functional results in 243 patients]. AB - STUDY AIM: The aim of this retrospective study was to evaluate the 38 month results of laparoscopic fundoplication for gastroesophageal reflux disease (GERD). PATIENTS AND METHODS: Two hundred forty three consecutive patients were operated laparoscopically. The surgical procedures were complete fundoplication with division of short vessels (Nissen: 80 patients), without division of short vessels (Nissen-Rossetti: 68 patients) or partial fundoplication of 270 degrees (Toupet: 95 patients). The mean follow-up was 38 months. Functional results were evaluated in 225 patients (92.5%) using a questionnaire with visual analog scales. RESULTS: The morbidity rate was 5%, higher after Nissen procedure (6.5%). With a follow-up of 3 months: a dysphagia coted 5/10, a gas bloat syndrome coted 4/10 and colon distension present in 61% of patients, were significantly more frequent after Nissen procedure. GERD recurred early in 4.5% of patients. With a follow-up of 38 months: dysphagia rate (coted 1/10) was significantly higher after Nissen. Dysphagia still persisted in 8 patients (9%) after Nissen. Colon distension and flatulence were more present after Nissen fundoplication. GERD recurrence rate was 12%. Pyrosis was significantly higher after Toupet fundoplication. Continuous medical treatment was necessary in 19 patients (8%). The satisfaction of patients was coted 7.5/10 without difference between to the three types of fundoplication. CONCLUSION: The total laparoscopic fundoplication for GERD seems to be a safe and efficient operation. This procedure proves to be more effective than partial fundoplicature but with a grater morbidity. Whatever the type of fundoplicature, the satisfaction of patients was good. PMID- 11933632 TI - [Laparoscopic re-establishment of digestive continuity following Hartmann's procedure. Retrospective study of the French Society of Endoscopic Surgery]. AB - STUDY AIM: Reversal of the Hartmann's procedure is associated with a high morbidity. The aim of this study was to evaluate the feasibility and results of laparoscopic reversal of the Hartmann's procedure. PATIENTS AND METHOD: Thirty eight consecutive patients, mean age 60 +/- 13.5 years were included in this retrospective study. The most common indication for the primary procedure was diverticular disease (70%). The mean time from the primary operation to the reconstruction was 136 +/- 124 days. The stoma was first dissected in 24 patients, allowing introduction of the first port. In the remaining 14 patients a standard umbilical port was inserted. The amount of adhesions was classified as low in 13 patients, mild in 15 patients and severe in 10 patients. All patients had a mechanical anastomosis. RESULTS: The conversion rate was 15%, due to adhesion problems in 5 patients and for a positive leakage test in one. The morbidity rate was 23.5% including 8 surgical complications. One patient died after post operative peritonitis complicating an anastomic leakage. The average hospital stay was 10 +/- 4.4 days. CONCLUSION: Our results indicate that laparoscopic reversal after Hartmann's procedure is feasible. The morbidity is lower than after classical open reconstruction. The presence of diffuse peritonitis at the primary operation as well as a short delay before the reconstruction, are important factors of conversion. PMID- 11933633 TI - [Operable lung cancer and synchronous adrenal masses: role of laparoscopic adrenalectomy combined with pulmonary resection]. AB - STUDY AIM: Assessment of laparoscopic adrenalectomy in the management of operable non-small cell lung cancer (NSCLC) associated with solitary and synchronous adrenal mass. PATIENTS AND METHODS: In a retrospective study, we reviewed 3 patients with operable NSCLC proved by pulmonary biopsy and an isolated synchronous adrenal mass shown by abdominal CT scan. We first performed a laparoscopic adrenalectomy followed by pulmonary resection. RESULTS: All patients had a laparoscopic adrenalectomy without any conversion or treatment-related death. Hospitalization stay ranged from 5 to 6 days. A left pneumonectomy has been performed immediately after this first hospitalization in 2 cases and after a first cycle of chemotherapy in the third case. Pathologic examination showed a NSCLC adrenal metastasis in 2 cases and an adrenocortical adenoma in the last case. During the follow-up 2 patients died of other distant metastasis and a mediastinal lymph node recurrence has been diagnosed in the third patient, actually treated by a second line chemotherapy. CONCLUSION: Despite those bad results that concern patients T3 M+ in 2 cases, laparoscopic adrenalectomy seems to be very interesting in selected cases. Considering that pulmonary resection can be done after, it represents a mean of diagnosis at least better than fine needle aspiration biopsy. Laparoscopic adrenalectomy might also be considered in the resection of a synchronous and isolated metastasis as a way to improve survival. PMID- 11933634 TI - [Giant parathyroid tumors: characterization of 26 glands weighing more than 3.5 grams]. AB - AIM OF THE STUDY: To determine the characteristics of giant lesions of the parathyroid glands weighting more than 3.5 g. PATIENTS AND METHOD: Twenty-six patients operated on between 1989 to 2001 were included in this retrospective study. Anatomical, biological, clinical and histological parameters were analyzed. Data were compared both with a personnal series of the last 220 patients with hyperparathyroidism operated on in our department and with the results of a primary hyperparathyroidism multicentric study conducted by the French Association of Surgery (AFC). RESULTS: They were 14 females and 12 males with a mean age of 58.57 +/- 13.72 years (ranged: 26-80). Mean weight of the parathyroid glands was 9.87 +/- 9.76 g (ranged: 3.5-40). The diagnosis of parathyroid disease was suspected by symptoms and incidentally discovered hypercalcemia in 17 and 8 cases respectively. In one case, the adenoma was misdiagnosed as a thyroid nodule. Mean calcemia was 125.42 +/- 19.6 mg/L, mean phosphoremia was 21.6 +/- 6.9 mg/L, mean seric parathormone concentration was 451.44 +/- 530.18 ng/L. Comparing with our personnel 220 HPT-series, they was no statistically difference concerning the mean age, but number of males and biological measurements were significantly higher in presence of a giant adenoma. Comparing with the study of the AFC group, there was no statistically difference concerning the symptoms, especially in regard to the asymptomatic forms discovered by hypercalcemia and to the acute hypercalcemia forms. Minor ectopic localizations were found in half of the cases. All the glands were considered as beginnings. After surgery, one patient had a severe hypocalcemia in relation to a hungry bone syndrome. CONCLUSION: Giant adenomas have no specific symptoms even if functional status seems to be more active. Diagnosis is made during the sixth decade as usual. Male people are more often concerned. At surgery ectopic localizations are present in 50% of the cases. In our study their size is not a sign of malignancy. After surgery severe hypocalcemia can occur if a long past of bone disease exists. PMID- 11933635 TI - [Parathyroid cyst. Report of ten cases]. AB - AIM OF THE STUDY: To evaluate the characteristics of the parathyroid cysts (PC). PATIENTS AND METHOD: Ten patients with PC were included in this retrospective study. The PC were discovered as follows: cervical mass (n = 3), hyperparathyroidism (n = 3), incidentally during thyroid surgery (n = 3) and screening for obesity (n = 1). Intracystic parathormone determination was performed after fine needle aspiration in 2 cases. RESULTS: Mean cyst measurements were 27 mm (ext: 5-70 mm) to 22 mm (5-45 mm). Nine cysts were cervical (resection by cervicotomy), and one was mediastinal (resection by sternotomy). In addition to the resection of the PC, 3 adenomas, 1 hyperplasia of the parathyroid glands and 3 benign thyroid diseases were recognized and treated during the cervicotomies. CONCLUSION: The diagnosis of PC is not common and must be based primarily on the study of the cyst liquid obtained by percutaneous puncture (intracystic parathormone measurement). PMID- 11933636 TI - [Complications of misdiagnosed Bochdalek hernia in adults. Literature review]. AB - AIM OF THE STUDY: To report 3 new cases of complicated Bochdalek hernia (BH) in adulthood and to review the literature about this rare condition. CASE REPORT: Three adult patients were operated on for a BH undiagnosed until occurrence of acute complication. The first patient, 27 years-old, had small bowel obstruction and the diagnosis of BH, unrecognized on chest X-ray, was established on barium meal and CT scan. The second patient, 38 years-old, had epigastric pain and gastric obstruction: diagnosis of BH, unrecognized at a previous laparotomy, was established on CT scan and barium enema. The third patient, 88 years-old, had respiratory failure and gastric obstruction: diagnosis of BH, unrecognized on chest X-ray, was established on CT scan. The 3 patients were operated on through laparotomy (n = 2) ou thoracotomy (n = 1) with one post-operative death. DISCUSSION: In adulthood, BH can remain asymptomatic for a long time before occurrence of a acute digestive or respiratory complication. Chest X-ray can be normal or misinterpreted. CT scan seems to be the most reliable examination to diagnose BH. CONCLUSION: In adulthood, diagnosis of BH should be evocated in case of respiratory or upper digestive symptoms. PMID- 11933637 TI - [Hibernoma: a rare case of massive weight loss]. AB - Hibernoma is a rare soft tissue benign tumor composed of cells similar to those of brown fat observed in fetus and hibernating animals. Brown fat has thermogenous properties, by the way of carbohydrates and lipid catabolism, and can be of an important mean in thermoregulation. A massive weight loss is a rarely reported sign in patients with hibernoma. We report herein the case of a 47 man with a history of isolated weight loss, of 16 kg over 4 months. Clinical examination has shown a swelling of the right flank. Surgical resection has been made and histopathological examination has shown hibernoma. The post-operative weight gain confirmed the relationship between hibernoma and weight loss. PMID- 11933638 TI - [Intrascrotal herniation of the ureter]. AB - We describe one case of intra-scrotal hernia of the right ureter. The anomaly was recognized on a pre-operatory urogram. It was an extra-peritoneal ureteral hernia. There are two types of ureteral hernia: the para-peritoneal hernia with a peritoneal sac; the extra-peritoneal type with only a fatty hernia. The mecanism of the latter illustrated by our case is a prolapse of the retro-peritoneal fat. It is not possible to practice an urogram before the cure of every inguinal hernia, so the prevention in this rare situation of the damage of ureter is to be cautious in the resection of huge lipomas and sliding fat when operating on inguinal hernias. PMID- 11933639 TI - [Retro-cost-xyphoid hernia and common mesentery: a case diagnosed in an adult]. AB - Authors analysed Morgagni's hernia pathogenesis through a new case associated to a common mesentery position and revealed by respiratory distress after delivery. PMID- 11933640 TI - [Usefulness of intraoperative 123I-MIBG during reoperation for malignant pheochromocytoma: a case report]. AB - Intraoperative MIBG radionuclear scanning has been used to improve pathological foci localization and surgical accuracy in patients with neural crest derived tumors. This intraoperative detection has been reported in less than 10 patients during reoperation for pheochromocytoma. We report a case of 123I-MIBG intraoperative detection allowing to improve surgical resection quality during reoperation for pheochromocytoma. The use of intraoperative MIBG radionuclear scanning is helpful when reoperation for pheochromocytoma is performed. PMID- 11933641 TI - [Surgery technique: preventive ligation of the thoracic duct during esophagectomy for cancer]. AB - Chylothorax is a rare but life-threatening complication of esophagectomy for cancer. Elective ligation of the thoracic duct above the diaphragm does not suppress completely the risk of chylothorax due to possible trauma of the thoracic duct wall at the level of ligation, or incomplete ligation in case of anatomic variation. This study describes a technique of preventive ligation "en block" with surrounding tissues in order to minimize the risk of chylothorax following oesophagectomy, whatever performed transthoracically or through transhiatal approach. PMID- 11933642 TI - [Impact of obesity on postoperative results of laparoscopic elective colectomy for sigmoid diverticulitis: prospective study]. PMID- 11933643 TI - [Response to Doctor G. Foucher]. PMID- 11933644 TI - [What is your diagnosis? Lichen ruber (lichen planus)]. PMID- 11933645 TI - [Associated symptoms and relevant associated illnesses in idiopathic Parkinson syndrome]. AB - Besides the four cardinal symptoms tremor, rigidity, akinesia and postural instability Parkinsonian patients may suffer from a number of associated symptoms which can have a negative influence on quality of life and which may require specific therapeutic measures. As it is to be expected in elderly people, patients with Parkinson's disease may suffer from other pathologies requiring medical treatment. In order to avoid undesired interactions the various treatments have to balanced carefully. PMID- 11933646 TI - [Autonomic disorders in idiopathic Parkinson syndrome: diagnostic relevance and therapeutic possibilities]. AB - Involvement of the autonomic nervous system in Parkinson's disease (PD) demonstrates the multiple system character of this disease exceeding the extrapyramidal system. Forty to sixty percent of patients with PD suffer from symptoms of autonomic failure impairing their quality of life. Autonomic failure in PD is caused by damage to the postganglionic part of the autonomic nervous system. Scintigraphy with radiolabeled metaiodobenzylguanidin (MIBG) provides evidence of autonomic involvement at an early stage of the disease and enables an early differential diagnosis of PD versus other neurodegenerative disorders. Sensitivity to identify PD versus multiple system atrophy is 89.7%, specificity in 94.6%. Orthostatic hypotension is a frequently overlooked symptom in PD. Diagnosis and cause of orthostatic hypotension can be identified with clinical methods. There are effective physical and pharmacological treatments to improve symptoms. PMID- 11933647 TI - [Fatigue, daytime somnolence and sleep disorders in Parkinson patients]. AB - Many patients suffering from Parkinson's disease complain about chronic fatigue and daytime somnolence. During the last few years attention has been drawn to "sleep attacks", which are supposed to be due mainly to dopamine agonists. Sleep disturbances during the night are quite frequent. It is important to search for the probable causes in each individual case in order to be able to install an efficacious treatment. PMID- 11933648 TI - [Psychological and cognitive problems in Parkinson disease--therapeutic possibilities]. AB - Depression, hallucinations, psychosis and cognitive deficits may often complicate advanced Parkinson's disease. Their detection and treatment have extraordinary importance, as they may cause significant invalidity and even an increase in mortality. Optimization of antiparkinsonian therapy may exert a positive influence on depressive symptoms, and should therefore be performed before antidepressant drugs are started. On the other hand, hallucinations and dementia do usually benefit from a discontinuation or dosage reduction of anticholinergic drugs, selegiline, DA-agonists and amantadine. When a levodopa monotherapy is indicated, slow-release formulations should be avoided. When a neuroleptic treatment is needed, clozapine and maybe quetiapine should be preferred. Preliminary evidence suggests that cholinesterase inhibitors might partially improve cognitive deficits in Parkinson's disease. PMID- 11933649 TI - [Effect of associated symptoms and associated illnesses on treatment strategy in idiopathic Parkinson syndrome]. AB - Treatment of Parkinson's disease (PD) is primarily focussed on motor symptoms. During the course of PD and with advancing age comorbidity as well as nonmotor PD symptoms become increasingly important. If these factors go beyond short disturbances of well-being, therapeutic consequences must be drawn. In these cases modifications of PD treatment on the one hand and treatment of associated symptoms or comorbidity on the other hand should be considered. PMID- 11933651 TI - Three-dimensional power Doppler in study of embryonic vasculogenesis. AB - Three-dimensional power Doppler sonography plays an important role in obstetrics, predominantly for assessing the relationship between fetal vascular system and surrounding organs. Presenting volume data in a standard anatomic orientation is valuable for assisting both ultrasonographers and pregnant patients to recognize anatomy more readily. This investigation was designed as an observational cross sectional study. A group of 25 patients in gestational age of five to eleven weeks were recruited for the study. Architecture of the embryonic vessels was depicted by 3D power flow mapping and analyzed visually. Vascular 3D measurements were done through 3D color/power histogram and expressed by Vascularization Index (VI) and Vascularization Flow Index (VFI). Volume of the embryo increased exponentially throughout the observation period. The VI and VFI showed no change despite an exponential growth of embryonic volume. The findings of our study suggest that in cases of physiologic embryonic vasculogenesis there is a homeostatic steady state between tissue demands and blood supply given through the vascular network. PMID- 11933650 TI - Two-dimensional and three-dimensional ultrasound of fetal anasarca: the glass baby. AB - Fetal edema was recognized in the past as a complication of alloimmune disease. More recently, fetal edema is frequently seen in conditions that cause non-immune fetal hydrops with increased soft tissue thickness. Classically there is a halo pattern around fetal head, neck, thorax, and abdomen. Fetal edema is associated with karyotype abnormalities, with multiple congenital anomalies, and with certain fetal infections like parvovirus B19 that cause severe fetal anemia. In the present case there was no evidence of infection or karyotype abnormality, but there was hypoplasia of umbilical cord vessels, pulmonary hypoplasia, and pericardial effusion. The etiology of fetal hydrops may remain unknown in up to 30% of cases. PMID- 11933652 TI - The assessment of placental blood vessels by three-dimensional power Doppler ultrasound. AB - OBJECTIVE: This study was performed in order to compare the performance of classical two dimensional (2D) and three dimensional (3D) ultrasound, both with power Doppler technology, in the visualization of the placental vascular network during ongoing pregnancy. METHODS: 15 pregnant volunteers in the third trimester of pregnancy were examined by 2D and 3D power Doppler ultrasound. The aim of the study was to follow the branching of the main stem vessel as far as possible distally in the placenta. In addition, we assessed the visualization rate of terminal parts of uteroplacental circulation, radial and spiral arteries. RESULTS: There was no difference in the visualization of primary placental stem vessels by 2D and 3D power Doppler. However, 3D power Doppler performs better distally, with statistically significant differences at the level of secondary stem (p = 0.03), and even more prominent differences at the level of tertiary stem vessels (p = 0.0008). There was no difference in the visualization rate of radial and spiral arteries (p > 0.05). CONCLUSION: We found 3D superior to 2D ultrasound with power Doppler technology in the determination of the distal vascular branches of the fetal placental blood vessels. PMID- 11933653 TI - Assessment of early chorionic circulation by three-dimensional power Doppler. AB - Three-dimensional power Doppler sonography is a unique instrument that enables assessment of vascular signals within the whole investigated area. Hemodynamical changes included in the process of early placentation are one of the most exciting topics in investigation of early human development. This investigation was designed as an observational cross-sectional study. A group of 25 patients in gestational age five to eleven weeks were recruited for the study. After acquirement of the volume containing three-dimensional power Doppler data of the pregnant uterus, the signals belonging to the chorion were isolated. Vascular 3D measurements were undertaken through 3D color/power histogram and expressed by Vascularization Index (VI) and Vascularization Flow Index (VFI). Volume of the chorion increased exponentially throughout the observation period. The VI and VFI positively correlated with the crown-rump length and chorion volume, and showed gradual increment through the investigation period. This investigation produced results confirming gradual augmentation of the loci and intensity of the intervillous flow in pregnancies between five and eleven gestational weeks. PMID- 11933654 TI - Limb deformities and three-dimensional ultrasound. AB - AIM: To assess the ability of three-dimensional (3D) ultrasonography for improvement of antenatal detection of limb deformities. METHODS: 347 patients were selected from a routine outpatient clinic or sent for supervision from other units because of suspected anomalies of fetal extremities. 3D ultrasound devices used in the study were Combison 530D and Voluson 530D MT (Kretztechnik, Zipf, Austria) with a 3-5 MHz annular array transducer for three-dimensional volume scanning. RESULTS: In 41 of 347 patients the initial diagnosis was suspected by two-dimensional sonography (gestational age 18-32 weeks). In 28 of 41 suspected cases the diagnosis of abnormalities was determined after examination by 3D sonography: 17/28 clubfoot, 3/28 hand-polydactily, 2/28 upper limb contractures, 1/28 lower limb contractures, 4/28 micromelia within the syndrome of skeletal dysplasia. In 13 of 41 suspected cases, normal anatomy was confirmed using 3D sonography. CONCLUSION: Three-dimensional sonography is the "method of choice" for the detection of an isolated defect of a single limb, developmental or positional deformations and minor defects of hands and feet. Surface-mode reconstruction of the complete limb and transparent-view reconstruction of the entire skeletal structure are effective technical advantages enabling a completely new visual perception of the unborn baby. PMID- 11933655 TI - The assessment of fetal brain morphology and circulation by transvaginal 3D sonography and power Doppler. AB - OBJECTIVE: The objective of this article is to describe the use of transvaginal 3D ultrasound in prenatal neuroimaging and to investigate its clinical usefulness. METHODS: Firstly, 18 fetuses with hydrocephalus, ventriculomegaly and/or space occupying lesion were examined by transvaginal 3D sonography, and fetal CNS abnormalities were evaluated. Multiplanar image analysis and volumetric assessment were performed off-line. Longitudinal volume changes of target structure were evaluated, and usefulness of transvaginal 3D ultrasound was evaluated. Secondly, in 56 normal cephalic fetuses of between 18 and 32 weeks, 3D Doppler volume acquisition and reconstruction of the intracranial vascular structure were performed. RESULTS: Longitudinal changing appearance in the same cutting section of the enlarged ventricle or cystic lesion could be demonstrated in all cases and volumetric assessment was also successful in all cases. Longitudinal objective data were useful in the brain assessment, consultation and counselling. 3D Doppler acquisition time was 5.6-26 seconds and symmetrical 3D reconstruction was successful in 51.8%. CONCLUSION: Transvaginal 3D imaging technology provides us with not only comprehensive intracranial images in exactly the right sections, but also with objective volume data. Prenatal information with objective data analyses is useful in consultation for both specialists and parents, and leads to proper management of CNS diseases. PMID- 11933656 TI - 3D ultrasound and perinatal medicine. PMID- 11933657 TI - Prenatal diagnosis: what does four-dimensional ultrasound add? AB - INTRODUCTION: The key benefit of 4D ultrasound lies in providing real-time 3D images of embryonic or fetal movements, previously limited by technological advancement. AIM: To classify types of first trimester embryonic and fetal movements in normal pregnancies as seen by 4D sonography. RESULTS: Three types of movements can be visualized in the first trimester: gross body between seven and eight weeks, limb movements after ten weeks and complex limb movements after 11 weeks of gestation. DISCUSSION: An alteration from the given pattern of motoric development should be considered as an indication for further investigation. CONCLUSION: 4D ultrasound enables visualization of more details of the dynamics of small anatomical structures. Therefore, body and limb movements can be visualized a week earlier than with 2D. PMID- 11933658 TI - Three-dimensional sonoembryology. AB - Three-dimensional (3D) ultrasound plays an important role in obstetrics, predominantly for assessing fetal anatomy. Presenting volume data in a standard anatomic orientation valuably assists both ultrasonographers and pregnant patients to recognize the anatomy more readily. Three-dimensional ultrasound is advantageous in studying normal embryonic and/or fetal development, as well as providing information for families at risk for specific congenital anomalies by confirming normality. This method offers advantages in assessing the embryo in the first trimester due to its ability to obtain multiplanar images through endovaginal volume acquisition. Rotation allows the systematic review of anatomic structures and early detection of fetal anomalies. Three-dimensional ultrasound imaging in vivo compliments pathologic and histologic evaluation of the developing embryo, giving rise to a new term: 3D sonoembryology. Rapid technological development will allow real-time 3D ultrasound to provide improved and expanded patient care on the one side, and increased knowledge of developmental anatomy on the other. PMID- 11933659 TI - Three-dimensional (3D) ultrasound--a useful imaging technique in the assessment of neonatal brain. AB - Clinical application of ultrasound began about fifty years ago. From one dimensional A-mode, through two-dimensional real time and Doppler examinations, a new era in clinical ultrasonography then began in the late eighties. Development of computer technology enabled introduction of 3D ultrasonography into clinical practice. In obstetrics ultrasound revolutionized fetal follow-up, but it was as important for the detection of intracranial pathology during the neonatal period and infancy. Two-dimensional real time ultrasonography was [table: see text] an exciting method that changed our understanding of the prevalence and pathophysiology of brain pathology in premature and term infants. Will application of 3D ultrasonography bring any substantial improvement to neuroimaging diagnostics in the newborn period? This article attempts to find the answer to this question, despite the limitations set by the short period of application of 3D neurosonography in neonates. The advantages of 3D brain ultrasonography application in neonates are: quicker and observer independent data acquisition, the possibility of off-line data analysis, projection of 3D data on a 2D plane with volumetric, color and power Doppler studies. Unavailability of equipment is the main reason why 3D ultrasonography was performed in only half of the newborns in whom it was indicated. Cost of equipment prevents introduction of 3D as a standard diagnostic procedure in neonates, although its diagnostic value is indisputable. PMID- 11933660 TI - Events from ovulation to implantation studied by three-dimensional ultrasound. AB - During the last decade transvaginal color Doppler has played an important role in increasing understanding of early human development. Although our knowledge of early pregnancy development has recently improved, little is known about the most critical period of human development: between conception and implantation. Recent advances in 3D ultrasound have made studies of follicular development, ovulation and uterine receptivity more accurate. The same method can be used for evaluation of the Fallopian tube patency and assessment of the ovarian and uterine causes of infertility which hamper processes of early human development. Storage capacities, reconstruction of the volume images and simultaneous viewing of all three orthogonal planes are the main advantages of this method. Introducing 3D ultrasound into assessment of early pregnancy has enabled visualization and volume estimation of the gestational sac, yolk sac and embryo. Switching on power Doppler superimposed to 3D gray scale can detect early vasculogenesis within the intervillous space and embryo/fetus. Here we review the potential application of this novel technique in monitoring the morphological and functional processes from ovulation towards implantation and early pregnancy. PMID- 11933661 TI - Screening for uterine abnormalities by three-dimensional ultrasound improves perinatal outcome. AB - The main goal of our study was to assess the true incidence of surgically correctable uterine abnormalities (congenital uterine anomalies, submucous leiomyoma, endometrial polyps and intrauterine synechiae) in the infertile population attending our tertiary infertility clinic. All of the infertile patients enrolled in the study were evaluated by three-dimensional ultrasound. Another objective was to assess pregnancy rates before and after operative hysteroscopy in patients affected by uterine causes of infertility. Good quality 3D images were obtained in all 3850 infertile patients, and in 23.2% of them 3D US revealed surgically correctable uterine abnormalities. The incidence of uterine septum in our general infertile population was 17.9%. Uterine septum was the most common uterine abnormality accounting for 77.1% of the intracavitary lesions. Out of 310 patients that were followed-up, 225 (72.6%) patients achieved pregnancy. The rate of term deliveries after septal incision was 57.7%, while 15.4% of patients had preterm deliveries. The rate of spontaneous abortions dropped from 41.7% before, to 11.9% after hysteroscopic resection of the septum. Three-dimensional ultrasound can be used as a screening method for detection of uterine abnormalities in patients suffering from infertility. We found significant improvement in reproductive outcome after operative hysteroscopy in secondary infertile patients with septate uterus and/or other uterine abnormalities. PMID- 11933662 TI - Multiple congenital contractures (congenital multiple arthrogryposis). AB - Arthrogryposis, is the occurrence of joint contractures of variable etiology that start prenatally. Arthrogryposis may result from neurologic deficit, neuromuscular disorders, connective tissue abnormalities, amniotic bands, [figure: see text] or fetal crowding. Arthrogryposis may result from no apparent hereditary causes (neuropathic, for example) or may be the result of hereditary factors (myopathic form, for example). Ultrasound diagnosis depends on observation of scant or absent motion of fetal extremities. Prognosis depends on the specific etiology of the contractures. PMID- 11933663 TI - [Early surgery of immature hemangiomas with the aid of an ultrasonic scalpel. Apropos of 81 cases]. AB - BACKGROUND: The natural course of immature hemangiomas in infants is well-known. A rapid phase of growth from 6 to 8 months is followed by a period of stability then regression. Since approximately 70% of these immature hemangiomas resolve spontaneously, abstention is generally the rule. The volume or localization of certain lesions may nevertheless have a serious functional or morphological impact. MATERIAL AND METHODS: This retrospective study included 81 children who underwent surgery between October 1994 and March 2000. The children were aged 2 to 38 months at the time of surgery. Orbital localizations predominated (33 children). The indication for surgery was based on two criteria: risk of a functional impairment or risk of morphological sequela. All children with orbital hemangiomas with a functional risk of amblyopia were initially treated with corticosteroids. Surgery was performed in case of failure. The CAVITRON was used for 77 children and DISSECTRON for 4. These two ultrasound devices allowed easy dissection with little hemorrhage. RESULTS: There were no peroperative hemorrhagic complications. Few postoperative complications were observed. After resection of the orbital hemangiomas there was little functional impact and the postoperative ophthalmologic examinations were normal within several weeks. Mean follow-up was 12 months after surgery. Use of an ultrasound dissector allowed early and safe treatment of immature hemangiomas. DISCUSSION: Certain voluminous or poorly localized hemangiomas, particularly on the face, can have a serious function, morphological or psychological impact. Medical treatment is not always active and surgical resection may be required before the development of definitive sequelae. Ultrasound dissection, not previously used in this indication, can contribute significantly to the surgical outcome as demonstrated in these children operated on early. This technique is safe and shortens operative time. In light of these results, we believe early resection of immature hemangiomas should be reevaluated. It should not be considered as a last resort but rather as a complementary treatment. PMID- 11933665 TI - [Systemic infections of dental origin]. AB - Microorganisms harbored in the oral cavity have long been recognized to cause systemic disease with a well known mechanism of spread via the blood stream. Different factors, including the presence of periodontal disease, the number of dental extractions or the duration of dental surgery have an influence on the risk of bacteremia. Infectious endocarditis is classically cited, but there are other potential problems including brain abscess, meningitis, lung or lie abscess, and even for some authors, a risk of thrombotic heart disease. It is important to keep in mind that some of these focal infections may be possible complications of dental (or buccal) infection. Prevention is crucial. PMID- 11933664 TI - [Evaluation of mandibular movement after condylar fracture]. AB - BACKGROUND: The purpose of this work was to compare results of two treatments for fracture of the mandibular condyle: functional therapy and intermaxillary block. MATERIAL AND METHODS: Forty patients were assessed: 32 unilateral fractures and 8 bilateral fractures. Half of the patients were given functional treatment and half had intermaxillary fixation for 40 days. Mouth opening amplitude, protrusion/retraction, and lateral movements were measured at inclusion, day 15, 30 and 180 for patients in the first group and at inclusion and on day 180 for the blocked patients. RESULTS: Movement amplitude at the end of treatment was always greater in patients given functional treatment. This difference was found for all types of movement. And was even more marked in patients with bilateral fractures. Occlusion was restored in all patients given intermaxillary fixation, but 2 patients in the functional treatment group had an occlusal impairment due to insufficient compliance with the treatment protocol. DISCUSSION: Functional treatment is more adapted than intermaxillary fixation for fractures of the mandibular condyle, particularly for bilateral fractures. Careful surveillance is nevertheless required to avoid malocclusion. An endobuccal device may be helpful in selected cases. PMID- 11933666 TI - [The Revue de Stomatologie et de Chirurgie Maxillo-Faciale, official journal of the Swiss Society of Maxillo-Facial Surgery]. PMID- 11933667 TI - [Oral self-mutilation and congenital analgesia: diagnosis and therapeutic problems. Apropos of a case]. PMID- 11933668 TI - [Temporomandibular joint pain-dysfunction syndrome: danger!]. PMID- 11933669 TI - [Radial forearm or Chinese flap]. AB - The Chinese or radial forearm flap, first described by Yang et al. (1981), is useful both as a free or a pedicle flap. This flap has proved to be one of the safest and most versatile flap in widely differing clinical situations. It is generally classified as a fasciocutaneous flap, but can also be used together with a segment of radius as a composite flap. However, the most common objection to this flap is the donor-site morbidity. The donor defect is significant, with a poor graft take over the exposed tendons. Inclusion of bone in the flap leads to a greater contour defect and to a risk of fracture. Many suggestions are given to minimize the complications on the donor-site defect. The surgical technique, the advantages, the disadvantages, and at least the indications of this flap are detailed. PMID- 11933670 TI - [Nasal injuries during labor and in early childhood. Etiopathogenesis, consequences and therapeutic options]. AB - Childhood and perinatal nasal traumatisms involve an anterior septal deviation or an anterior septal lysis. These complications induce a soft nasal tip. Nasal obstruction et oral ventilation are responsible for the development of facial and occlusal sequelae. A better knowledge of anatomy and physiopathology of nasal traumatisms is needed for an earlier treatment. PMID- 11933671 TI - [Optimized Practical Teaching and the Tour of France: a necessary alliance?]. PMID- 11933672 TI - [Pleomorphic adenoma of the parotid gland]. PMID- 11933673 TI - [Rouen 2001: an excellent start]. PMID- 11933674 TI - [The ethics of research: impact factors]. PMID- 11933675 TI - Benzo[1,2-c]1,2,5-oxadiazole N-oxide derivatives as potential antitrypanosomal drugs. Structure-activity relationships. Part II. AB - The preparation of new derivatives of benzo[1,2-c]1,2,5-oxadiazole N-oxide is described. These derivatives were chosen in order to investigate and confirm previous structural features found necessary to display an adequate antitrypanosomal activity. The compounds synthesized were tested in vitro against epimastigote forms of Trypanosoma cruzi. The presence of a bromine atom in the benzo system produced compounds less active than the corresponding de-halo analogues. However, 5-(bromomethyl)-7-bromobenzo[1,2-c]oxadiazole N-oxide (23) was the most cytotoxic compound against T. cruzi. For this, the 50% inhibitory dose (ID50) was determined, it was of the same order as that of Nifurtimox. From statistical analysis we could establish a relationship between lipophilic hydrophilic balance of the derivatives with their effectiveness as antichagasic compounds. PMID- 11933676 TI - Azaberbene derivatives as alpha 2A-adrenoceptor antagonists. AB - The synthesis of octahydro-8H-isoquino[2,1-g][1,6]naphthyridine derivatives 2a-f was carried out. These new compounds are selective alpha 2A-adrenoceptor antagonists. PMID- 11933677 TI - Investigation of antiulcer activities of imidazo[1,2-alpha]pyridinyl-2 alkylaminobenzoxazoles and 5,6,7,8-tetahydroimidazo[1,2-alpha]pyridinyl benzoxazoles with electron-topological (ET) method. AB - The study presents structure-activity considerations of a series of imidazo[1,2 alpha]pyridiny-2-alkylaminobenzoxazoles(I) and 5,6,7,8-tetahydroimidazo[1,2 alpha]pyridinylbenzoxazoles(II) investigated for anti-stress ulcer activity with the electron-topological method. A series of 39 compounds including 24 active and 15 weakly active was studied. It is shown that the fragment determined by the electron-topological method in an active molecule is responsible for anti-stress ulcer activity. Quantitative structure-activity relationships with electron topological approach of these compounds are discussed in terms of the statistical program STATGRAF-7.0. PMID- 11933678 TI - Synthesis and cytotoxicity of 9-alkoxy-1,5-dichloroanthracene derivatives in murine and human cultured tumor cells. AB - 9-Alkoxy-1,5-dichloroanthracenes were successfully prepared. Their cytotoxicity was evaluated in vitro on rat glioma C6 cell lines and human hepatoma G2 cell lines, respectively. Alkylation of 1,5-dichloro-9(10H)-anthracenone with either the appropriate alcohols or alkyl chlorides in the presence of sulfuric acid or sodium hydride, respectively, furnished this structural class of anthracenes. Contrary to mitoxantrone, cytotoxic properties were observed as documented by the reactivity of the novel compounds and potent in vitro activity against C6 cells and hep G2 cells over a wide range of structural variants. Among these compounds, 5c, 5h, 5l and 5n are potent cytotoxins. They inhibit C6 cell growth in culture, indicated by using 2,3-bis(2-methoxy-4-nitro-5-sulfophenyl)-2H-tetrazolium-5 carboxanilide sodium salt (XTT) colorimetric assay. By using this assay it was also shown that 5c, 5d and 5l possess potent cytotoxicity on hep G2 cells. The most active compound displaying in vitro cytotoxicity was the 9-butoxy derivative 5h with IC50 values 0.02 microM against C6 cells, as compared with mitoxantrone with IC50 values 0.07 microM. The most active compound displaying in vitro cytotoxicity against hep G2 cells was 5c with IC50 values 1.7 microM (mitoxantrone: 0.8 microM). Structure-activity relationships (SAR) of these compounds with respect to the nature of the alkoxy substitution in the 9 position are discussed for both cell lines. PMID- 11933679 TI - Sigma receptor binding profile of a series of analgesic tropane derivatives. AB - The binding profile on sigma receptor subtypes of a family of tropane derivatives, previously studied as analgesic and nootropic drugs, is reported. The results show that the sigma receptors are not involved in the enhancement of acetylcholine release and of the consequent analgesic activity of this class of drugs. Structure-affinity relationships for sigma receptor subtypes are discussed. They lead to the identification of a few useful leads for further development of potent sigma ligands within the series. PMID- 11933680 TI - Synthesis of imidazo[2,1-a]phthalazines, potential inhibitors of p38 MAP kinase. Prediction of binding affinities of protein ligands. AB - Based upon molecular modeling, the pharmacophore of potential inhibitors of p38 MAPK (mitogen-activated protein kinases) is discussed and the predictive binding affinities are calculated. Syntheses of original diarylimidazo[2,1-a]phthalazines obtained by Suzuki coupling are described. PMID- 11933681 TI - Advances in the management of endometrial adenocarcinoma. A review. AB - Endometrial adenocarcinoma is the most common and curable gynecologic neoplasm; the five-year survival for women with surgical stage I disease ranges from 83% to 93%; stage II, 73%; stage III, 52%; and stage IV, 27%. The absence of an asymptomatic latency phase amenable to detection through screening and the already excellent cure rates seen with early-stage disease have precluded the need for endometrial cancer screening programs. Adenocarcinomas constitute 97% of endometrial cancers, with endometrioid the most common histologic subtype. Two different pathways of endometrial carcinogenesis exist. One arises in a background of estrogen excess, giving rise to atypical hyperplasia as the malignant precursor of the more common endometrioid adenocarcinomas. The use of oral contraceptives has consistently been shown to decrease the risk of developing endometrial carcinoma via this pathway, with 12 months or more of continuous use decreasing the lifetime risk by 40-50%. The alternate pathway of endometrial carcinogenesis represents malignant transformation of atrophic endometrium and proceeds through endometrial intraepithelial carcinoma as the malignant precursor of the more virulent serous papillary and clear cell endometrial adenocarcinomas. The staging of endometrial cancer (according to the International Federation of Obstetrics and Gynecology) is surgical. Recent studies suggest a therapeutic benefit associated with extensive retroperitoneal lymph node evaluation to determine the disease extent and thereby more effectively direct potentially life-saving adjuvant therapy. Adjuvant radiation therapy, known to have survival benefit in advanced-stage disease, may also have survival benefit in intermediate-risk surgical stage I disease on the basis of results recently released from a Gynecologic Oncology Group study. The use of radiation therapy, systemic chemotherapy and hormonal therapy, alone or in combination, is recommended for primary advanced and recurrent disease. PMID- 11933682 TI - ICAM-1 in maternal serum and amniotic fluid as an early marker of preeclampsia and IUGR. AB - OBJECTIVE: To determine if plasma and amniotic fluid levels of intercellular adhesion molecule-1 (ICAM-1) at 16 weeks' gestation could be predictive of preeclampsia or intrauterine growth retardation (IUGR). STUDY DESIGN: A retrospective analysis was undertaken in 44 serum samples stored for Down's syndrome screening at 16 weeks' gestation and 44 amniotic fluid samples obtained by midtrimester amniocentesis. RESULTS: No significant difference was found between women who subsequently developed preeclampsia or IUGR and the control group. CONCLUSION: This study failed to demonstrate that ICAM-1 may be an early serum marker of preeclampsia or an amniotic fluid marker of IUGR. PMID- 11933683 TI - Clinical and patient estimation of fetal weight vs. ultrasound estimation. AB - OBJECTIVE: To compare clinical and patient estimation of fetal weight to ultrasound estimation. STUDY DESIGN: Prospective study of clinical, patient and ultrasound estimation of fetal weight at term. RESULTS: A total of 200 pregnant women participated. There was no statistically significant difference between clinical and sonographic estimates of fetal weight: of the estimates, 64.0% were within 10% of the actual birth weight vs. 62.5% (P > .2). There was no statistically significant difference between patient and sonographic estimates of fetal weight: of the estimates, 53.5% were within 10% of the actual birth weight vs. 62.5%, respectively (P < .1). Senior resident clinical and sonographic estimates of fetal weight were superior to junior resident estimates: 75.2% of clinical estimates were within 10% of the actual birth weight vs. 59.2% (P < .03), and 73.1% of sonographic estimates were within 10% of the actual birth weight vs. 58.3% (P < .05). Nulliparous and multiparous patients were equally accurate in estimating fetal weight: 48.1% of estimates were within 10% of birth weight vs. 57.4% (P > .2). CONCLUSION: Sonographic estimation of fetal weight offers no advantage over clinical or patient estimation of fetal weight at term. Senior resident clinical and sonographic estimates are superior to junior resident estimates. Parity has no effect on patient accuracy in estimating fetal weight. PMID- 11933684 TI - Laser-assisted intracytoplasmic sperm injection in human oocytes. AB - OBJECTIVE: To explore potential applications of a non-contact, 1,480-nm diode laser to intracytoplasmic sperm injection (ICSI) of human oocytes. STUDY DESIGN: Human oocytes obtained from in vitro fertilization (IVF) patients and failed to fertilize 24-48 hours after conventional IVF were used for ICSI along with discarded sperm. A noncontact, 1,480-nm diode laser was employed to immobilize sperm, to open a hole in the zona pellucida (ZP) and to perform ICSI through the hole. After ICSI and its simulation, oocytes were examined for formation of pronuclei, cleavage and normality of the cytoskeleton. RESULTS: The 1,480-nm diode laser permitted fast and easy sperm immobilization and microdrilling of ZP to facilitate microinjection. Of the 78 injected oocytes, 53 (68%) survived the procedure, and 13 (25%) of them formed two pronuclei by 18 hours. Further culture of two fertilized eggs resulted in cleavage up to the eight-cell stage before cease of culture. Four oocytes were fixed after simulation of the procedure without injecting sperm. None of them showed gross abnormalities in cytoskeletal organization. CONCLUSION: A noncontact, 1,480-nm diode laser can be used for the immobilization of sperm and for opening a hole in the ZP to facilitate ICSI, biopsy manipulation toward preimplantation genetic diagnosis and assisted hatching. PMID- 11933685 TI - Ultrasonographic observation of uterine contractility. New perspectives. AB - The possibility of noninvasively assessing uterine contractility in ultrasound scans allows improvements in our understanding of hormonal regulation and of the influence of uterine contractility on the human embryo implantation process in natural and controlled ovarian hyperstimulation cycles. Ultrasound is a reliable tool for studying contractions of the nonpregnant uterus and confirms that uterine contractility is influenced by ovarian hormones and indicate that it partakes in the embryo implantation process in humans. PMID- 11933686 TI - Laparoscopic uterine suspension by round ligament plication. AB - OBJECTIVE: To describe a method for laparoscopic uterine suspension by round ligament plication using standard suturing instruments. STUDY DESIGN: Forty-six women underwent uterine suspension for treatment of chronic pelvic pain due to uterine retroversion over a period of five years. All operations were performed by the principal author. RESULTS: Forty-six patients treated with this technique were selected from 388 who underwent clinical evaluation and diagnostic laparoscopy for chronic pelvic pain, defined as lasting more than six months. There were no intraoperative complications in this series. Mean follow-up was 23.6 months (range, 6-42). At each follow-up, patients were asked to rate their pain relative to preoperatively as unchanged, minor, moderate, significant or resolved. At six months, 44 of 46 patients (96%) reported at least some improvement, with 17 (37%) reporting significant improvement, and 18 (39%) reporting resolution of pain. Sixteen of 27 patients (59%) at 24 months' follow up and 15 of 23 patients (65%) with 36 months' follow-up reported significant improvement in or resolution of their pain. CONCLUSION: For patients experiencing chronic pelvic pain associated with uterine retroversion, round ligament plication is an effective method of repositioning the uterus and reducing or eliminating the patient's symptoms. PMID- 11933687 TI - Interaction of pubertal status, mood and self-esteem in adolescent girls. AB - OBJECTIVE: To examine the relationship between pubertal status, mood and self esteem in adolescent girls. STUDY DESIGN: The symptoms of depression, stress, anxiety, stress perception and self-esteem in 971 girls from 8 to 16 years old were evaluated. The girls' pubertal stage was assigned according to Tanner staging. With the ANOVA test, the mean scores of the psychological symptoms were correlated with the girls' pubertal development. RESULTS: The mean age of the girls was 12.5 +/- 2.4, years (mean +/- SD). Three percent of them had symptoms of severe depression, 2.1% had symptoms of severe anxiety, and 20% demonstrated severe stress perception. The symptoms of depression, anxiety and stress perception were highest in girls at Tanner stage V (P < .001). However, the girls' self-esteem decreased as the Tanner stage increased (P < .001). CONCLUSION: The girls' pubertal development was associated with higher scores on depression, anxiety and stress perception and with poor self-esteem. These results may be explained by the modulating effect of sexual hormones on the girls' central nervous system. PMID- 11933688 TI - Obesity and second-trimester abortion by dilation and evacuation. AB - OBJECTIVE: To examine the relationship between body mass index (BMI) and dilation and evacuation (D&E) procedure characteristics. STUDY DESIGN: Data were collected on all women seeking elective surgical second-trimester abortion during a six week period at a free-standing abortion clinic. Information gathered included age, pregnancy history, height, weight, intravenous sedation, time for procedure, estimated blood loss, use of special instruments, amount of manual dilation needed and gestational age by ultrasound and pathology. Difficulty was rated by a single physician immediately following the surgery using a 10-cm line later converted to a visual analog score. RESULTS: One hundred ninety-eight women underwent D&E during the study period. Data were complete for 163 (82%). Subject demographics and procedure outcomes were analyzed with BMI as a variable. A trend was observed for increased procedure difficulty, time for procedure, blood loss and complications as BMI increased. On average, the procedure for a subject with BMI > or = 30 took 20% longer and was rated as 40% more difficult to perform as compared to procedures on women with BMI < 30. CONCLUSION: We identified a trend with BMI score and some D&E procedure characteristics, but the power of the study was limited. Abortion providers should be aware that obesity may necessitate the availability of special instruments and/or an alterations in surgical technique. PMID- 11933689 TI - Pelvic organ prolapse and connective tissue abnormalities in Korean women. AB - OBJECTIVE: To evaluate the relationship between pelvic organ prolapse in Korean women and joint hypermobility, which suggests a metabolic collagen fiber abnormality. STUDY DESIGN: Between March 1998 and March 2000, we investigated 55 patients with prolapse. The prevalence of joint hypermobility, by measuring finger extension angle, and the proportion of patients with joint hypermobility were measured in patients with pelvic organ prolapse and benign gynecologic patients (control group). RESULTS: In middle-aged women (40-59 years), the average finger extension angles were higher in the POP group than in the control group (50.04 +/- 9.70 degrees vs. 39.50 +/- 12.19 degrees, respectively; P < .05), but in older women there was no significant difference between the two groups (42.84 +/- 13.05 degrees vs. 43.00 +/- 13.34 degrees, respectively; P > .05). CONCLUSION: The prevalence of joint hypermobility was higher in the POP group and with advanced POP stage (III, IV) than in the control group and early POP stage (I, II). Our results suggest that intrinsic connective tissue abnormality is related to the development of pelvic organ prolapse. Further study involving more patients with pelvic organ prolapse is warranted, and molecular studies to determine the genetic basis of pelvic organ prolapse are also required to further elucidate this abnormality. PMID- 11933690 TI - Amniotic membrane grafting to treat refractory labial adhesions postpartum. A case report. AB - BACKGROUND: Labial adhesions are common in prepubertal children, but their occurrence postpartum is extremely rare. CASE: A woman developed labial adhesions after a vaginal delivery. Topical estrogen cream was ineffective. Simple excision was performed, but the adhesions reformed two weeks later. Finally, amniotic membranes were used to cover the labia minora after they were separated by excision. The wound healed without adhesion formation. CONCLUSION: Excision plus amniotic membrane grafting is effective for recurrent labial adhesions. PMID- 11933691 TI - Postcoital vaginal cuff rupture 10 months after a total vaginal hysterectomy. A case report. AB - BACKGROUND: Transvaginal evisceration following total vaginal hysterectomy secondary to coitus is extremely rare. CASE: A woman presented 10 months following a total vaginal hysterectomy with complaints of progressive postcoital abdominal and shoulder pain as well as a pinkish vaginal discharge. Examination revealed a 3-cm defect at the left edge of the vaginal cuff. Corrective surgery followed overnight observation with pain management. CONCLUSION: Postcoital vaginal cuff disruption is rare, and complications can range from bowel evisceration to hemorrhage. Management should be tailored to the severity of the complications. PMID- 11933692 TI - Extreme caudal agenesis. Possible drug-related etiology? AB - BACKGROUND: Caudal regression syndrome (CRS) is a rare anomaly of the lower body pole that represents a continuum of congenital malformations ranging from isolated sacral agenesis to absence of the lumbosacral spine and major visceral anomalies. While the exact etiology of this syndrome is unclear, maternal diabetes, genetic factors, teratogens and vascular anomalies altering blood flow have been hypothesized to play a role in its pathogenesis. CASE: A fetus had extreme hypotrophy of the caudal body pole, aplasia of the lower spine and complete renal agenesis diagnosed in the second trimester by ultrasound. Maternal history revealed the use of minoxidil solution for preventing hair loss for four years prior to and during gestation. Also, the mother had taken trimethoprim sulfamethoxazole during the first trimester for treatment of upper respiratory disease. No maternal diabetes or history of familial genetic diseases was evident. CONCLUSION: In an extreme form of CRS consisting of complete aplasia of the lower body pole and viscera and additional malformations, a possible drug related etiology was suggested but should be confirmed by more studies. PMID- 11933693 TI - Twin heterotopic pregnancy after assisted reproduction. A case report. AB - BACKGROUND: Heterotopic pregnancy is a potentially catastrophic form of ectopic pregnancy and is increasing in incidence secondary to assisted reproductive technology. Early diagnosis and intervention are important in avoiding short- and long-term morbidity. CASE: A 36-year-old, nulliparous woman became pregnant by in vitro fertilization and embryo transfer. A total of three embryos were transferred. She presented to the emergency room approximately six weeks after transfer with the complaint of severe abdominal pain. Laboratory analysis revealed a decreasing hematocrit with stable vital signs despite continued abdominal pain. On transvaginal ultrasound, two fetal poles were present, with cardiac activity in two of the three gestational sacs. At surgery the patient was found to have a ruptured tubal pregnancy in addition to the intrauterine gestations. Pathologic analysis revealed a twin tubal pregnancy. CONCLUSION: Heterotopic pregnancy should be considered in the differential diagnosis of any patient who becomes pregnant by assisted reproduction techniques and presents with signs and symptoms of ectopic pregnancy. PMID- 11933694 TI - [Effect of muscle contraction on heart rate and skin blood flow. Preliminary results]. AB - Evaluation of heart rate variability is an important tool to study several diseases. The most important tests include the variability of the heart rate as a function of time and as function of frequency, variability of blood pressure and pulsatile cutaneous blood flow (PCBF). We studied the effect of a sustained contraction of the quadriceps muscle on the instantaneous heart rate and pulsatile cutaneous blood flow in 10 healthy subjects. We measured the R-R interval and the instantaneous heart rate, in the time domain, 10 beats before the contraction, the R-R interval that coincided with the onset of the contraction, and the 30 R-R intervals after the beginning of the contraction. To measure the PCBF we used a photopletismograph designed for this purpose. We recorded the surface electromyographic activity (EMG) of the quadriceps muscle to determine the onset and end of the muscle contraction, and measured respiratory movements with a belt. We found an increase of the instantaneous heart rate at the first and second beats after the onset of the contraction. There was a decrease of the pulsatile cutaneous blood flow of the finger pad immediately after the contraction, which lasted for approximately eight beats. The findings support the idea of a central command and co-activation of the motor and sympathetic nervous systems. This efferent sympathetic activity seems to act not just on the contracting muscle but also on the heart and the cutaneous blood vessels of the extremities. These findings imply that it is quite likely that the sympathetic efferent effect has a supraspinal origin. This is a test that could be used to study cardiovascular control by the sympathetic nervous system. PMID- 11933696 TI - [Early arterial reflexion and ventricular extrasystole. A novel mechanism detected with sphygmokinetocardiography]. AB - ANTECEDENT: Through sphygmokynetocardiography (SKCG) an exploratory method that records an electrocardiographic signal, a carotid pulse (CP), and two vibriograms (kinetocardiograms) of the left ventricle (LV) recorded in the left hemithorax (anterior kinetocardiogram, AKC) and the subcostal region of left abdomen (posterior kinetocardiogram of PKC, vibrations transmitted through the hemidiaphragm) we observed a systolic precocious reflection wave (Rw) in the CP and prolongation of LV ejection time (LVET) measured in AKC or in PKC of the previous sinusal pre-extrasystolic beat (PEB) vs control beats (CB) in cases with ventricular extrasystoles (VEs). OBJECTIVE: To demonstrate whether the intervals just mentioned are associated with ventricular extrasystoles. METHOD: Sixty cases: 30 with VEs, group A, and 30 without arrhythmia, group B, were studied through SKCG. The LVET and the arterial reflection index or ARI = Ta-rw/LEVT, Ta rw = time between initial ventricular impulse to reflexive wave, were measured. RESULTS: Demography was similar in both groups. PEB had a longer LVET than the CB (291 +/- 41 vs 279 +/- 39, p < 0.01) and ARI was shorter (0.36 +/- 0.17 vs 0.58 +/- 0.21, p < 0.001). CONCLUSIONS: 1) Distention of the LV due to Rw, possibly through the well-known experimental mechanism of electromechanic feedback, is believed to underlie the arrhythmia. 2) The observation has important clinical implications. PMID- 11933695 TI - Balloon angioplasty in aortic coarctation: a multicentric study in Mexico. AB - OBJECTIVES: To analyze immediate and long-term results of balloon dilation for aortic coarctation in a three-center experience in Mexico, and to determine factors associated with increased risk. BACKGROUND: Results demonstrated that the procedure is effective and safe, however its use in some groups is still controversial, specially in neonates and infants. METHODS: In a ten-year period, 333 patients with aortic coarctation on underwent balloon dilation with an immediate success rate of 93.7% and a major complication incidence of 1.8%. Of the total cohort, 272 patients were followed for a period of 24.3 +/- 20 months. Demographic and procedural data were analyzed to determine factors related to a poor outcome or to sustained high blood pressure. RESULTS: Cox regression analysis found age (risk ratio 3.42 p = 0.0001), isthmic hypoplasia (risk ratio 4.64, p < 0.0001), and post-dilation gradient (risk ratio 2.19, p = 0.0113) as independent risk factors for a follow-up event, mainly restenosis. Age at dilation was the only independent factor related to sustained hypertension with a seven-fold increase in the risk. CONCLUSIONS: Balloon dilation is an effective and safe alternative to treat aortic coarctation. Patients younger than one year of age, with severe isthmic hypoplasia and a post-dilation gradient > 20 mmHg have the highest risk to develop an event in the follow-up period. When the dilation procedure is performed in patients older than 10 years of age, and specially those older than 20 years, the probability that they remain or develop high blood pressure is increase. PMID- 11933697 TI - [Immediate and mid-term results following coronary stent implantation in diabetic patients]. AB - INTRODUCTION: We analyzed the clinical and angiographic outcome, including follow up, in diabetic, patients treated with percutaneous transluminal coronary angioplasty (PTCA) and stenting. MATERIAL AND METHODS: Between January 1995 and December 1999, 770 patients were treated with PTCA, 137 of them with diabetes (DM), 17.7% received 160 stents, average 1.16 stent/patient. Mean age was 58.70 +/- 8.44 years (range 35 to 80 years) and there were 92 men and 45 women. According to the clinical presentation, there were 54% with severe angina based on criteria from the Canadian Cardiovascular Society (CCS). Sixty two percent of patients had prior myocardial infarction and 68% had multivessel disease. Mean vascular obstruction was 90 +/- 6.3% and the ejection fraction was 45.3 +/- 8.7%. RESULTS: The immediate angiographic success was 94.4% in patients with DM. There were complications in 8.4% (acute myocardial infarction, and thrombosis). The average follow-up was 10 mouths (range 6 to 36) in 94% of the cases. The mortality was 4.3%. CONCLUSIONS: These results show that PTCA and stenting in patients with DM is a successful procedure, with few clinical and angiographics complications. PMID- 11933698 TI - [Disclosure of "Brugada's syndrome" with intravenous propafenone]. AB - We present the case of a woman with familial antecedents of a brother who died suddenly at the age of 39 years, and an other with Brugada syndrome. The ECG had not shown the typical pattern of the Brugada syndrome. Based on these antecedents, we decided to perform a pharmacological test with endovenous propafenone, which revealed the electrocardiographic alterations at 10 min after the administration. This is the first case in our knowledge in which endovenous propafenone manifested a concealed Brugada syndrome. PMID- 11933699 TI - Detection of right ventricular myocardial perfusion and contractile reserve by contrast echocardiography and low dose dobutamine in myocardial infarction after successful right coronary angioplasty. AB - We assessed right ventricular myocardial reperfusion through intracoronary contrast and transesophageal echocardiography after right coronary angioplasty. Low dose dobutamine was used to demonstrate contractile reserve in areas with microvascular integrity. PMID- 11933700 TI - [Repair of interatrial septal defect with an Amplatzer device. Experience with 3 cases]. AB - We performed transcatheter closure of an atrial septal defect (ASD) using an Amplatzer device in three patients, 2 women and 1 male child, aged 12, 54, and 4 years, respectively, coursing with ostium secundum ASD. Two with left to right shunt and the third with bidirectional shunt. The transesophageal echocardiogram revealed ASD with diameters of 13, 15, and 10 mm, the diameter with expanded catheter balloon was of 30, 26, and 17 mm, respectively. The superior border of the atrial septum was bigger than 5 mm in all three patients, whereas the inferior border could not be found through echocardiography in the patients aged 12 and 4 years. Amplatzer devices of 30, 26, and 17 mm were implanted, the inferior border of the first two could be supported on the aortic wall and the superior border on the septum. We observed a 100% occlusion in these two cases through ecocolor-Doppler. In the third patient, an inferior border of 16 mm and a superior border of 8 mm were determined, achieving and immediate and fast installation of the device with 100% occlusion. All three patients were in stable conditions, asymptomatic and without shunt at their one-month follow-up as revealed by a trans-thoracic echocardiography. It is concluded that the Amplatzer device is technically easy to install and yields adequate results even in patients with ample ASD and without inferior border of the inter-atrial septum. PMID- 11933701 TI - [Postinfarction statins. An unquestionable benefit. A therapeutic strategy that can be improved]. AB - Atherosclerotic disease is etiopathogenically related to the presence of hypercholesterolemia. Physicians have never had until now a generation of lipid lowering drugs with a comparable degree of efficacy, potency and safety as statins. The most explored field with statins has been the post myocardial infarction period, since this type of patients is at high risk of new cardiovascular events and, as a consequence, can obtain a marked benefit from intervention. Accordingly, three large-scale clinical trials with statins have conclusively demonstrated a significant reduction in cardiovascular events and in mortality after myocardial infarction. Nevertheless, although there is convincing evidence that statins should be implemented after myocardial infarction, there are still a number of relevant questions open and under study. The first of them is the value of the so-called pleiotropic properties of statins (additional properties independent from cholesterol lowering), so important for plaque stabilization. In addition, there is uncertainty about how early treatment with statins has to be started after the acute phase. Another point we do not know is the goal of lipid intervention (the level of total or LDL-cholesterol to be achieved). In fact, there is still a debate on whether patients with average levels of cholesterol or even with low levels are to treated. Finally, but related to the previous question, it would be very interesting to determine whether high doses of statins, are necessary or can low doses be equally efficacious. In conclusion, statins are a landmark in cholesterol-lowering interventions after myocardial infarction. Probably, the majority of survivors should be treated. At present, active research is focused on delineating more precisely the manner in which these drugs are to be used to prolong life in these patients. PMID- 11933702 TI - [Septal myocardial fibrosis. An electro-histologic comparison]. PMID- 11933704 TI - [Research design: a brief methodologic review]. PMID- 11933703 TI - [Arterial hypertension in Mexico: results of the National Health Survey 2000]. AB - AIMS: Hypertension remains as a major cause of cardiovascular morbidity in Mexico. The Health National Survey 2000 of Mexico was performed to analyze the current status of the prevalence of some risk factors such as diabetes, hypertension (HTA), obesity, smoking, and proteinuria. METHODS: A National Survey was carried out in Mexico where 45,300 individuals between 20 to 69-y.o. were screened. The estimated sample size was calculated considering the total number of persons into the mentioned age; a minimal prevalence of 6% of the included risk factors, at a significance level of 0.05; a maximum relative error of 0.145, and a rate of response of at least 70%. Diagnosis of HTA was accepted in: previous medical diagnosis with prescription of antihypertensive drugs, or high blood pressure (> or = 140/90 mmHg) detected during the interview. Data were adjusted for the national distribution of age-groups and gender (established in 2000, INEGI). RESULTS: 38,377 (98.8%) individuals were correctly screened for blood pressure. The prevalence of hypertension in Mexico was 30.05% (34.2% in men and 26.3% in women). The prevalence was directly related with age and gender. The percentage of mexicans with HTA after 50-y.o. is > 50%. The prevalence was greater in women after 50-y.o. Among persons with hypertension, treatment was detected in 46% and approximately 20% of them were controlled (< 140/90 mmHg). The percentage of mexicans with HTA who were unaware that they have high blood pressure was 61%. The total percentage of HTA controlled was 14.6%. The North states had the greater prevalence (approximately 34%) of HTA while South states had the lower prevalence (27%). The odds ratio (age/sex-adjusted) for HTA were: for diabetes 1.54(CI95%, 1.44-1.63); for obesity 2.3 (CI 95%, 2.22-2.43); for smoking 1.26 (CI 95%, 1.21-1.32). For proteinuria subjects, 40% had HTA. CONCLUSIONS: Around 15 millions of mexicans between 20 to 69-y.o. had hypertension, 60% of them are unaware. The prevalence of hypertension in Mexico (30.05%) has increased. Among persons with hypertension -15% are controlled. The North of Mexico has the higher prevalence of hypertension. Diabetes, smoking, and obesity increase the risk of hypertension. The 2000 National Survey of Health shows the epidemiological trend in several important chronic diseases in Mexico and declare an urgent need of new strategies for detection, control and treatment of hypertension. PMID- 11933705 TI - The joy of (understanding) money. PMID- 11933706 TI - Improving your cash flow. PMID- 11933707 TI - Competing with heart. PMID- 11933708 TI - Protecting your lab system with anti-virus & firewall software. PMID- 11933709 TI - Marketing what's unique about your laboratory. PMID- 11933710 TI - What dentists say about lab marketing. PMID- 11933711 TI - What you don't know could kill your business. PMID- 11933713 TI - Forget retirement. PMID- 11933712 TI - Nine qualities that make up a good team! PMID- 11933714 TI - Sell, but stay in the saddle. PMID- 11933715 TI - Introducing orthodontics into the dental lab. PMID- 11933716 TI - Investing in CAD/CAM. PMID- 11933717 TI - Lab Manager 2000. Software package for small labs. PMID- 11933718 TI - Improving implant framework passive fit and accuracy through the use of verification stents and casts. AB - Passive fit of implant frameworks is related in part to the accuracy of the master cast. A method of fabricating a verification stent to check the accuracy of the master cast prior to waxing and casting the implant framework is discussed. The methods utilize routinely available materials and do not require any special instrumentation or equipment. The stent can be fabricated either by the laboratory or the dentist. PMID- 11933719 TI - Bad account or bad attitude? PMID- 11933720 TI - Personnel files: what should and should not be in them. PMID- 11933721 TI - Do you and your technicians work together as a team. PMID- 11933722 TI - Attracting certified dental technicians. PMID- 11933723 TI - An invitation to dental technology professionals worth five CDT scientific credits for being published in the journal of dental technology. PMID- 11933724 TI - What's bringing back our daughters and sons? PMID- 11933725 TI - Peter Stein's Pricing for Profit Workbook. PMID- 11933726 TI - Using an artificial neural network to predict healing times and risk factors for venous leg ulcers. AB - OBJECTIVES: This study aimed to identify the risk factors that influence the healing process of venous leg ulcers treated with compression bandaging, with a view to predicting healing time. METHOD: A retrospective cohort study was performed on data collected prospectively on 325 consecutive patients presenting with 345 venous ulcers at the Salford Primary Care Trust leg ulcer clinic between January 1997 and December 1999. Use of an artificial neural network (ANN) technique accurately predicted the healing times for 68% of the patients. RESULTS: The ANN demonstrated that healing was significantly related to a history of previous leg ulceration, 'quite wet' ulcer exudate, high body mass index, large initial total ulcer area, increasing age and male gender. CONCLUSION: The ability to identify at presentation ulcers that might be resistant to standard therapy would allow early consideration of more radical treatments such as hospitalisation, wound debridement or venous surgery. PMID- 11933728 TI - A histological comparison of acute inflammatory responses with a hydrofibre or tulle gauze dressing. AB - OBJECTIVE: This study analysed the physical properties of Aquacel hydrofibre dressing in rat partial-thickness wounds, focusing on the acute inflammatory infiltrate of granulocytes and macrophages in the wound and the dressing. METHOD: Partial-thickness wounds (2 x 2 cm) were made on the back of 60 anaesthetised male Wistar rats and covered with Aquacel (n = 30) and tulle gauze (n = 30). The rats were killed on postoperative days one, two, three, four, seven and 10 (10 animals per day and five per dressing). Re-epithelialisation and Polymorphonuclear (PMN), fibronectin and macrophage activity were then analysed. RESULTS: PMN leucocytes (granulocytes) were captured in the dressing and remained active there, resulting in a reduced number in the wounds when compared with tulle gauze. A fibrin layer formed between the dressing and the wound, creating a physical barrier. Macrophages infiltrated the wound bed but could not be detected in the dressing. Little inflammation was observed in the wound bed and the macrophages operated primarily in the repair mode. Active PMNs in the dressing provided an appropriate antimicrobial environment. Tulle materials became embedded in wounds and were associated with a more disturbed pattern of epithelial outgrowth. Aquacel stayed 'on top' of wounds, with only minimal incorporation into the superficial epidermis. CONCLUSION: The observations of the physical properties of different materials and their histological consequences correlate well with published clinical results, particularly in relation to the speed of re-epithelialisation and the level of scarring. PMID- 11933727 TI - Effect of bacteriological status on pressure ulcer healing in patients with advanced cancer. AB - OBJECTIVES: This study aimed to identify the quantitative and qualitative bacteriological status of pressure ulcers in patients with advanced cancer, and the effects on the healing process. METHOD: We recruited 34 patients with advanced cancer who had 38 superficial grade II and III pressure ulcers. Patients were randomly allocated to treatment with Lyofoam/polyurethane foam dressing (Seton) or Aquagel/hydrogel dressing (Wytw.Opatrunkow). Efficacy, treatment times and healing rates were noted. Qualitative studies were undertaken on 38 pressure ulcers and quantitative bacteriological studies on 19 ulcers. RESULTS: There was no statistical difference between the two treatment groups in efficacy, healing rates and treatment times. Bacteriological assessment identified 92 species. CONCLUSION: The bacteria did not cause any clinical signs of infection. Variations in the number and types of bacteria in individual wounds did not correlate to the grade of ulcer or the dressing used. PMID- 11933729 TI - Is there a role for sustainable medicine in wound care? AB - Is it ethical to prolong life by providing aggressive palliative care to chronically ill patients with a short life expectancy? Here, Andrea Cavicchioli argues that epidemiology studies can help us to resolve this difficult ethical dilemma. PMID- 11933730 TI - Modulating the hypermetabolic response to burn injuries. AB - A huge rise in metabolic rates occurs following a burn injury. Practitioners can alleviate this problem by controlling the patient environment and providing appropriate nutritional support. Drug therapy is likely to be a future option. PMID- 11933731 TI - Clinical trials in wound care I: the advantages and limitations of different clinical trial designs. AB - Clinical trials need to be robust if practitioners are to apply their recommendations into practice. This article describes the various research designs used in wound care and the pitfalls that may compromise the reliability and validity of the results. PMID- 11933732 TI - Evaluation of dermagraft as an alternative to grafting for open fasciotomy wounds. AB - Wounds produced following open fasciotomy can be healed by grafting but this requires a second operation. Is Dermagraft, a tissue-engineered human fibroblast derived dermis that can be applied on an outpatient basis, a viable alternative? PMID- 11933733 TI - [From the apprehension of sexually transmissible diseases to the prevention of HIV]. AB - Over the past few years in France, the incidence of human immunodeficiency virus (HIV) has not decreased. Among the most frequent sexually transmissible diseases (STD) in France (condyloma, genitoanal herpes, chlamydia infections), certain STD, considered as negligible, have reappeared: gonorrhoea and syphilis affect male homosexuals and, to a lesser degree, men and women whose epidemiological profile remains to be determined. The health organization is not in favour of associating STD with HIV in its anti-aids strategy. However, acute STD are not only indicator of habits at risk for HIV, but are also potent co-factors of its sexual transmission. Fighting against HIV without creating a dialogue on STD is a waste of time and efficiency. From our experience with the STD, anonymous and free screening and the inter-disciplinary health education centres, we recommend the following: improved screening for HIV and other STD: concomitantly whenever possible, less invasive, free or reimbursed STD sampling, reliable and standardized techniques (polymerisation chain reaction or PCR and derivatives), itinerant screening for STD for persons who do not consult; ensured early, medical, social and psychological care of HIV and STD, emphasising the importance of compliance to treatment and prevention; ensured easy access and low cost of the male and female condoms; renewal and diversification of health relays, particularly in the private sector; staff training on STD and their epidemiological novelty; insisting on a transversal (HIV-other STD, curative preventive, among others) and pragmatic approach (intervention studies resulting in local action); renewal of the information and advice for the public: information on the relationship between HIV and other STD, on the frequent STD that are lesser known, such as condyloma and chlamydia infections, emphasis on compliance to prevention measures (abstinence or use of condoms) during at least three months after a risk of HIV/STD transmission or after an STD has been identified and treated. The aim is to enhance the acceptation or the appropriation by the public of concomitant STD/HIV screening, prevention measures and treatments proposed. This can only be implemented after a change in the institutions', carers and public's attitude towards STD. PMID- 11933734 TI - [Are the principles of treatment of chronic osteitis applicable to the diabetic foot?]. AB - OBJECTIVE: The interest of the management of bone infections in the diabetic foot, inspired by the recommendations for the treatment of chronic osteitis, was assessed in this study. METHODS: Twenty bone infections in 17 diabetic patients with moderate to mild infections of the feet were confirmed by the results of X ray and/or scintigraphic studies and bone surgery biopsy cultures revealing one or more bacteria sensitive to standard osteitis treatment (rifampicine + fluoroquinolone). The patients had received this treatment per os for a median duration of 6 months (3 to 10 months). Clinical follow-up was carried out during a consultation at 1, 3 and 6 months during treatment and then by telephone every six months after the end of treatment. Clinical success was defined as the disappearance of any local sign of infection and by the absence of relapse during the post-treatment follow-up period. The evolution of the bone infection was also assessed by the results of a control conducted 3 to 6 months after initiation of the antibiotic treatment. RESULTS: At the end of the treatment, all signs of infection had disappeared in 15/17 patients (88.2%) and no relapse had occurred in 14 (82.3%) patients at the end of a median post-treatment period of 22 months (12 to 41 months). Resection of necrotic bone was performed at the same time as the bone biopsy in 2 patients. The median duration of hospitalisation was of 14 days (3 to 53 days). During the study, a multi-resistant germ was isolated in 4 patients (1 Pseudomonas aeruginosa, 3 Staphylococcus aureus). During the post treatment follow-up, 3 patients dies from causes unrelated to the infection treated. No serious adverse event was reported during the study. DISCUSSION: The results of this pilot study support the rationale of applying the treatment regimens of chronic osteitis to diabetic lesions of the feet, but are only applicable to comparable patients presenting with non-severe lesions of the feet. Moreover, the use of antibiotics with potent selection of resistance such as rifampicine and fluoroquinolone, requires that bone biopsies be taken, which is not easy in all the diabetic foot care centres. We are presently conducting a study to identify the sub-populations of diabetic patients who could benefit from such treatment. PMID- 11933735 TI - [Severe sickle cell anemia revealed in an adult by medullary necrosis]. AB - INTRODUCTION: Drepanocytosis, even severe, may only be discovered in adults further to severe complications. OBSERVATION: A 44 year-old woman was hospitalised for a severe vaso-occlusive crisis, revealing a drepanocyte syndrome with heterozygote S-beta zero thalassemia. Evolution was marked by severe anaemia, the aregenerative nature of which, uncommon during vaso-occlusive crises and the absence of Parvovirus B19 infection, led to the diagnosis of medullar necrosis. Evolution was rapidly improved after transfusion of erythrocyte concentrations and symptomatic treatment of the pain. COMMENTS: Medullar necrosis is a rare entity with multiple causes. In severe drepanocyte syndromes it is concomitant to a severe vaso-occlusive syndrome, resulting from medullar ischemia due to specific microvascular damage. PMID- 11933737 TI - [Malignant pleural mesothelioma. A rare tumor that has become a world-wide public health problem]. PMID- 11933736 TI - [Catamenial pneumothorax. Failure of videothorascopic treatment]. AB - INTRODUCTION: Catamenial pneumothorax occurs in women during the menses. It is frequently associated with diaphragmatic fenestrations or episodes of pleural endometriosis. OBSERVATION: We report the case of a patients who had presented 25 episodes of right pneumothorax despite treatment with videothorascopy initiated after the 10th episode. These episodes coincided with menstruation. Diagnosis of catamenial pneumothorax was evoked and thoracic surgery was indicated. Exploration of the pleural fossa did not reveal symphysis but 7 diaphragmatic orifices, which were sutured before conducting mechanical pleurodesis. No relapse of pneumothorax was reported following surgery. COMMENTS: Around one hundred cases of catamenial pneumothorax have been reported in the literature. They represent 2.8 to 5.6% of spontaneous pneumothroax in women. Diagnosis is evoked on the occurrence of recurrent pneumothorax appearing 24 to 72 hours after the onset of menstruation. The pathogeny is uncertain and several mechanisms are frequently evoked. In the prevention of relapses, treatment consists in closing the diaphragmatic orifices with thoracotomy, resection of eventual bubbles, and pleural symphysis. Videothorascopy does not appear adapted to treatment. PMID- 11933738 TI - [Malignant pleural mesothelioma. From diagnosis to prognosis]. AB - EPIDEMIOLOGY: The incidence of malignant pleural mesothelioma has constantly increased over the past forty years. The recent measures of ban on the use of asbestos and the long latency of this tumor after exposure means that its peak incidence can be foreseen for the years 2010-2020. DIAGNOSIS: Various health professionals are involved in the care of this tumor, which benefits equally from progresses in clinical and fundamental research. Some progress has been made in understanding its oncogenesis as well as its histopathologic analysis. PROGNOSIS: Malignant pleural mesothelioma symptoms are rapidly invalidating and the patient's prognosis is bad at short-term. However, hope may come from the detection of early stages of the disease and from the individualization of good prognosis factors, permitting the selection of patients for whom some curative therapies are in course of evaluation. PMID- 11933740 TI - [The benefits of new immunosuppressive treatments in adult kidney transplantation]. AB - NEW KNOWLEDGE: Over the last few years, improved knowledge of the immunological mechanisms underlying transplant rejection have resulted in the development of new immunosuppressive agents capable of selectively blocking various steps of the immune response. NEW MOLECULES: The review focuses on new immunosuppressive drugs that have been developed for clinical use in renal transplantation and mechanism of action, advantages and side effects will be discussed for each of them. Neoral is a cyclosporin microemulsion, characterized by more consistent absorption. Tacrolimus shares with cyclosporin an ability to inhibit calcineurin but presents some differences. Mycophenolate mofetyl selectively inhibits purine synthesis and lymphocyte proliferation; it is easy to use and has been found effective in large clinical trials. Rapamycin and RAD are two related molecules that inhibit signal transduction by cytokines to T-cells, their use in transplant therapy remains to be determined. Anti-interleukin-2 receptor antibodies are also reviewed; they are easy to use and have been found effective. NEW STRATEGIES: These new immunosuppressive drugs provide new approaches in transplant therapy to improve their efficacy and safety. PMID- 11933739 TI - [Malignant pleural mesothelioma. Present data and perspectives for treatment]. AB - CHEMOTHERAPY: With regard to the efficacy of mono-chemotherapy and according to the literature, no cytotoxic substance, apart from methrotrexate at high doses, leads to a response rate of more than 20%. With regard to cyrotoxic associations, the published results show slightly betterresponse rates. IMMUNOTHERAPY: Interleukine 2 as well as various interferons have been tested alone or in association with chemotherapy. Fairly encouraging response rates have been reported. However, the possibility of severe adverse events must be taken into account. RADIOTHERAPY: The interest of prophylactic parietal radiation following invasive thoracic treatment has been demonstrated. Palliative use of radiotherapy is possible for pain, and more rarely for decompression. With curative aim, the results of isolated radiotherapy are disappointing. SURGERY: Palliative surgery is aimed at reducing the tumour and pleural symphysis. Curative surgery consists in wide extrapleural pneumonectomy, permitting total resection of the visceral pleura, or decortication pleurectomy leaving the lung in place. Mean survival of patients having undergone surgery is of 10 to 17 months with rates between 10 and 30% at 2 years ASSOCIATED THERAPY: For the first time, despite high morbidity rates, associated therapy has led to prolonged survival, whereas isolated therapy has not. This is the case with an association of radical surgery, radiotherapy and adjuvant chemotherapy. IN THE FUTURE: In the treatment of malignant pleural mesothelioma, genetic, anti-proliferative and immune therapy, that attempt to use the immune system of the patient to obtain an anti-tumour cytotoxic reaction, appear promising. PMID- 11933741 TI - ["More and better"!]. PMID- 11933742 TI - [Scientific evidence and medicine]. PMID- 11933743 TI - [You're anxious? How bizarre!]. PMID- 11933744 TI - [The Academy of Medicine worries about spreading use of cannabis]. PMID- 11933745 TI - [Medical demography, an overview]. PMID- 11933746 TI - [Futility in geriatric medicine, a real issue]. AB - Is futility age related? Several studies clearly demonstrate that outcome in critically ill elderly patients admitted to an intensive care unit, is more closely related to severity of illness than to age, which has little influence on prognosis. The outcome is closely associated with the severity of the illness, and age is of little influence. However, age based discrimination persists despite current recommendations from medical societies that age should not be used as a criteria for admission to intensive care. The concept of medical futility is influenced by individual or collective subjective values. Very old patients often are excluded from studies. It is difficult to apply evidence-based medicine in older people as results must be extrapolated from data in younger subjects. Thus, the true benefit of treatment is difficult to assess in the elderly. The image of the older patient, as perceived by younger health care professionals and society has a considerable impact on medical care. Medical futility is often based on clinical concepts which may lead to legal policies. It is influenced by social and economic factors. Health care professionals have a crucial role but their position, in close proximity to the patient, and its resulting emotional burden can be uncomfortable. In geriatric medicine, there is a special relationship between patients, relatives, and health professionals which requires genuine communication in order to optimise patients case. In conclusion, a balanced approach that avoids both overtreatment and therapeutic nihilism is important in order to provide old patients the best possible care. PMID- 11933747 TI - [A new antiviral for genito-anal condylomas]. PMID- 11933749 TI - [Colorectal spirochetosis, a possible but unusual cause of chronic diarrhea?]. AB - INTRODUCTION: Human intestinal spirochetosis has been known since the end of the nineteenth century. OBSERVATION: We report one case of intestinal spirochetosis revealed by chronic diarrhoea and diagnosed on colonic biopsies. The chronic diarrhoea disappeared with antibiotherapy. COMMENTARIES: The prevalence of intestinal spirochetosis in rectal and colonic biopsies, among patients with digestive disorders, varies in occidental countries from 2% to 7%. Presently, the pathogenic role of intestinal spirochetosis is controversial. PMID- 11933748 TI - [Functional intestinal disorders: assessment and perception by physicians of their patients perception. Results of two national studies]. AB - OBJECTIVE: In 1999, a first national survey on functional intestinal disorders (FID) revealed the patients' good perception of the medical corps and treatments proposed. The aim of the present study was to assess the physicians' perception of their patients, their symptoms and their degree of satisfaction. METHODS: The survey was conducted among the physicians who had recruited the patients of the initial study. Eight hundred and eighty five replies were analysed (general practitioners--GPs: 88%; gastro-enterologists--GE: 11%). RESULTS: The mean number of patients with FID seen in one week was of 11.40 (GP: 10.80; GE: 16.29). Thirty percent of patients were seen for the first time by a GP and 65 percent by a GE. The mean yearly number of consultations for these patients was of 7.16 for the GPs and 3.81 pour les GEs. A coloscopy was prescribed by 49% of GPs and 66% of GEs. The pain and transit disorders mentioned in 98 and 78.7% of cases had evolved over a mean of 56.57 months. COMMENTS: Although aware of the repercussion of FID in daily life, the practitioners evaluated a psychological component of 69% in the expression of the disorder. They considered the pain intense in 6% of cases versus the 41% expressed by the patient. The scores out of 10 for 8 items showed a mean result of 6.86 in their evaluation of the degree of the patients' understanding of their disease and its treatment, and of 6.13 for the quality of the medical act. These same criteria, assessed one year earlier by the patients themselves, showed better scores. PMID- 11933750 TI - [Post-traumatic cerebral thrombophlebitis]. PMID- 11933751 TI - [Effects of hormone replacement therapy in menopause on cardiovascular risk. Need for a European study]. AB - The increased risk of coronary heart disease (CHD) after menopause is currently attributed to estrogen deficiency. Many epidemiological (case-control and prospective) studies have reported a decreased risk (0.5-0.7) of CHD in postmenopausal women receiving hormone replacement therapy (HRT). However, there are some discordant studies, among which the Framingham study. Moreover, the observational studies were subject to several biases that may have falsely elevated the apparent benefit of estrogen: women taking estrogen tend to be wealthier, more educated and healthier than untreated women. Large randomized clinical trials of secondary (HERS, WEST) or primary (WHI) prevention have not confirmed cardioprotection with HRT. However, these studies used orally administered estrogens, while the non-oral route of administration is frequently used in Europe. From a biological point of view, estrogen has multiple effects that would be expected to be cardioprotective, including favorable changes in lipids, endothelial function, vascular reactivity and blood flow. However, concerning hemostasis factors, a pro-coagulant effect can be induced by the first pass liver effect of estrogen when administered orally, which is not observed with the non oral routes of administration. In addition, the synthetic progestin medroxy-progesterone acetate (MPA) inhibits the beneficial effects of estrogen on the arterial wall, whereas natural progesterone does not. It is therefore urgent that Europeans undertake a European "HERS study" in order to investigate the possible beneficial effect of non-oral estrogens (administered percutaneously or transdermally) associated with natural progesterone. PMID- 11933753 TI - Systemic anaphylaxis induced by intradermal testing. AB - We report the case of a patient who was referred for suspected anaphylaxis with anesthetic drugs. Gallamine was diagnosed the responsible drug and our patient developed anaphylaxis during intradermal testing (IDT) with this compound. IDT should be monitored carefully. PMID- 11933752 TI - [Role of pathological delayed-type hypersensitivity in chronic fatigue syndrome: importance of the evaluation of lymphocyte activation by flow cytometry and the measurement of urinary neopterin]. AB - Chronic fatigue syndrome or benign myalgic encephalomyelitis has been extensively described and investigated. Although numerous immunological abnormalities have been linked with the syndrome, none have been found to be specific. This article describes the detection of delayed-type hypersensitive responses to certain common environmental antigens in almost fifty per cent of patients with this syndrome. Such hypersensitivity can be detected by the intradermal administration of antigens derived from commensal organisms like the yeast Candida albicans, and then monitoring for a systemic reaction over the following six to forty eight hours. This approach can be consolidated by performing lymphocyte activation tests in parallel and measuring in vitro T-cell activation by Candida albicans antigens by three-colour flow cytometry based on CD3, CD4 and either CD69 or CD25. Another useful parameter is the kinetics of neopterin excretion in the urine over the course of the skin test. The results showed that the intensity of the DTH response correlated with the number of T-cells activated in vitro. Various factors have been implicated in the fatigue of many patients, notably lack of sleep. However, it remains difficult to establish causality in either one direction or the other. This work is in the spirit of a multifactorial approach to the group of conditions referred to as "chronic fatigue syndrome". PMID- 11933754 TI - [Delayed hypersensitivity to heparins and heparinoids]. AB - Delayed hypersensitivity to heparins and heparinoid is a problem for prophylaxis of thrombo embolic diseases. The hirudins did not seem to have any cross reactivity with the two others groups of anticoagulants. We present two clinical cases of delayed type reactions to heparins and heparinoid and we reviewed the literature about adverse reactions to low molecular weight heparins and the alternative possibilities. PMID- 11933756 TI - Challenges for medical informatics as an academic discipline. Proceedings of a workshop. March 2001, Madrid, Spain. PMID- 11933755 TI - A case control study of dermatophagoides immunotherapy in children below 5 years of age. AB - BACKGROUND: In this study we investigate the effects of parenteral immunotherapy on children below 5 years of age in order to prove the safety and clinical efficacy of dust mite specific immunotherapy (SIT) in such young patients. In young children, allergy is often due to dermatophagoides and causes a persistent inflammation that leads to recurrent respiratory infections. This condition impairs the normal development of airways by inducing remodelling. Although many paediatric allergists are used to treating young allergic children with SIT, this method is still subject to discussion. The benefits versus risks of SIT must also be defined. METHODS: In a case control study, 28 patients with dermatophagoides induced asthma and rhinitis treated with subcutaneous immunotherapy were assessed alongside a comparable control group. Symptom and drugs scores were collected from diary cards. RESULTS AND DISCUSSION: Scores proved to be significantly lower in the immunotherapy group in respect to the control group (p = 0.001). In particular, asthma attacks in the case group decreased significantly during the first year of treatment (p = 0.001). SIT was well tolerated and compliance was good. CONCLUSIONS: Our results confirm the safety and the efficacy of SIT in children below 5 years of age and above all, they confirm that SIT can begin at a very early age. PMID- 11933757 TI - Medical informatics: searching for underlying components. AB - OBJECTIVE: To discuss unifying principles that can provide a theory for the diverse aspects of work in medical informatics. If medical informatics is to have academic credibility, it must articulate a clear theory that is distinct from that of computer science or of other related areas of study. RESULTS: The notions of reusable domain antologies and problem-solving methods provide the foundation for current work on second-generation knowledge-based systems. These abstractions are also attractive for defining the core contributions of basic research in informatics. We can understand many central activities within informatics in terms defining, refining, applying, and evaluating domain ontologies and problem solving methods. CONCLUSION: Construing work in medical informatics in terms of actions involving ontologies and problem-solving methods may move us closer to a theoretical basis for our field. PMID- 11933758 TI - The micro-macro spectrum of medical informatics challenges: from molecular medicine to transforming health care in a globalizing society. AB - BACKGROUND: Medical informatics has always encompassed a very broad spectrum of techniques for clinical and biomedical research, education and practice. There has been a concomitant variety of depth of specialization, ranging from the routine application of information processing methods to cutting-edge research on fundamental problems of computer-based systems and their relations to cognition and perception in biomedicine. OBJECTIVES: Challenges for the field can be placed in perspective by considering the scale of each--from the highly detailed scientific problems in bioinformatics and emerging molecular medicine to the broad and complex social problems of introducing medical informatics into web related global settings. METHODS: The scale of an informatics problem is not only determined by the inherent physical space in which it exists, but also by the conceptual complexity that it involves, reinforcing the need to investigate the semantic web within which medical informatics is defined. RESULTS AND CONCLUSION: Bioinformatics, biomedical imaging and language understanding provide examples that anchor research and practice in biomedical informatics at the detailed, scientific end of the spectrum. Traditional concerns of medical informatics in the clinical arena make up the broad mid-range of the spectrum, while novel social interaction models of competition and cooperation will be needed to understand the implications of distributed health information technology for individual and societal change in an increasingly interconnected world. PMID- 11933759 TI - Integrating genomics into health information systems. AB - OBJECTIVE: To outline the main issues related to the impact of the data generated by the Human Genome Project on health information systems. A major challenge for medical informatics is identified, consisting of adapting traditional systems to new genetic-based diagnostic and therapeutic tools. METHODS: Reviewing and analysing the different health information levels from an organisational complexity point of view. A model is proposed to explain the interactions between health informatics, bioinformatics and molecular medicine. RESULTS: We suggest a new framework that integrates genetic data into health information systems. Using this model, new topics for future research and development are identified. CONCLUSIONS: We are witnessing the birth of a new era (post-genomics). In this era technological advancements in genomics offer new opportunities for clinical applications. Medical informaticians should play an important role in this new endeavour. PMID- 11933760 TI - Health care in the information society: what should be the role of medical informatics? AB - OBJECTIVE: To discuss the consequences for medical informatics in encouraging and advancing the development of information processing methodology (IPM) and information and communication technology (ICT) to contribute to high-quality and efficient health care. METHODS: Characterization of the current state of ICT, commenting on literature. RESULTS AND CONCLUSIONS: Medical informatics is the discipline, concerned with the systematic processing of data, information, and knowledge in medicine and health care. Our societies are continuously being influenced by modern IPM and ICT. It can be expected that these developments, leading us into an "information society", will continue. Three factors may significantly influence health care in the near future: the development of the population towards an aging society, progress in medicine, and progress in informatics. The major aims that will have to be achieved are the (1) patient centered use of medical data, (2) process-integrated decision support, using high quality medical knowledge, and (3) comprehensive use of patient data for clinical research and health reporting. Medical informatics research is needed on the electronic patient record, modern architectures for health information systems, and medical knowledge bases. In order to adequately pursue the goal of "transforming health care through innovative use of ICT for the 21st century", health care professionals are needed, who are well-trained in medical informatics, respectively health informatics. Medical informatics must offer such educational programs and assure a sufficiently high quality of education. PMID- 11933761 TI - Knowledge management to support performance-based medicine. AB - OBJECTIVES: To discuss research issues for medical informatics in order to support the further development of health information systems, exploiting knowledge management and information and communication technology to increase the performance of Health Care Organizations (HCOs). METHODS: Analyze the potential of exploiting knowledge management technology in medicine. RESULTS AND CONCLUSIONS: The increasing pressure on HCOs to ensure efficiency and cost effectiveness, balance the quality of care, and contain costs will drive them towards more effective management of medical knowledge derived from biomedical research. Knowledge management technology may provide effective methods and tools in speeding up the diffusion of innovative medical procedures. Reviews of the effectiveness of various methods of best practice dissemination show that the greatest impact is achieved when such knowledge is made accessible through the health information system at the moment it is required by care providers at their work sites. There is a need to take a more clinical process view of health care delivery and to identify the appropriate organizational and information infrastructures to support medical work. Thus, the great challenge for medical informatics is represented by the effective exploitation of the astonishing capabilities of new technologies to assure the conditions of knowledge management and organizational learning within HCOs. PMID- 11933762 TI - Medical informatics as a discipline at the beginning of the 21st century. AB - OBJECTIVES: To analyse the present situation of the discipline medical informatics and to propose actions for change. METHODS: Evaluation of the current situation mainly based on anecdotal evidence. RESULTS: The difference between the scientific and the engineering aspects of medical informatics get blurred. Because of the requirements of European funding medical informatics focuses more on engineering than on science. Too many manuscripts are submitted that describe engineered artefacts without a scientific purpose. Some of the subjects (like security issues) that are studied in medical informatics are not considered important by medical faculties thus impeding support. CONCLUSIONS: The methodological underpinnings of our research should be strengthened, impact studies should be more frequently performed; the quality of results reporting should be increased. PMID- 11933763 TI - Theory, abstraction and design in medical informatics. AB - OBJECTIVE: To analyze the scientific and engineering components of Medical Informatics. A clear characterization of these components should be undertaken to categorize different areas of Medical Informatics and create a research agenda for the future. METHODS: We have adapted a classical ACM and IEEE report on computing to analyze Medical Informatics from three different viewpoints: Theory, Abstraction, and Design. RESULTS: We suggest that Medical Informatics can be considered from these three perspectives: (1) Theory, from which medical informaticians formally characterize the properties of the objects of study, creating new theories or using and adapting existing theories (e.g., from mathematics), (2) Abstraction, from which medical informaticians deal with all aspects of medical information and create new abstractions, methods, and technology-independent models, which can be experimentally verified, and (3) Design, from which medical informaticians develop systems or act as information brokers or advisors between medical and technology professionals, to improve the quality of computer applications in medicine. CONCLUSION: Based on this framework, we suggest that Medical Informatics has an independent scientific character, different from other applied informatics areas. Finally, we analyze these three perspectives using data mining in medicine. PMID- 11933764 TI - Closing the loop between clinical practice, research, and education: the potential of electronic patient records. AB - OBJECTIVE: To discuss the possible contribution of electronic patient records in closing the loop among clinical practice, research and education. RESULTS AND CONCLUSIONS: Applying Information and Communication Technology (ICT) to a given medical domain is not merely adding a new technique. When introduced into an environment, ICT will initially often emulate or resemble the already existing processes. When workers and researchers in that domain begin to appreciate the potential of ICT, this initial stage is followed by more fundamental changes in that domain that take advantage of the potential of ICT. To understand the scope of the potential changes enabled by electronic records, three principle changes need to be understood. First, data recorded in computer memories can be readily retrieved and re-used for a variety of purposes. Second, once data are available in computer memories, the data can be transported easily. Third, as physicians (and patients) are using computers to record medical data, the same electronic record can be used to introduce other computer programs that interact with the user. New usage of data, however, generates additional requirements. Thus the experience in developing decision support systems and analyzing observational databases feeds back into the requirements for electronic medical records. Each patient-physician encounter, each investigation, each laboratory test, and each treatment in medical practice constitutes, in principle, an experiment. Ideally, we learn from each experiment. Electronic medical records will facilitate research that relies on data recorded in routine medical practice. The potential and challenge, however, of Medical Informatics lies in its ability to close the loop among clinical practice, research, and education. PMID- 11933765 TI - A UK operational practitioner view--some challenges of health informatics are trans-national. AB - OBJECTIVE: To explore the relevance of catalysts and inhibitors to the achievement of an inclusive identity for health informatics; particularly from an operational perspective in the UK. METHODS: Consideration of the different dimensions of health informatics, as practiced in a working healthcare delivery environment; specifically commenting on the synergy and disjunctions with academic and scientific practitioners in the some domain. RESULTS AND CONCLUSIONS: There appear to be common ground and internationally applicable issues across the domain. Recognizing the differences and similarities will contribute towards harmonisation of the field and its ultimate elevation to a mature discipline and profession. PMID- 11933766 TI - Challenges in medical informatics: perspectives of an international medical informatics organization. AB - OBJECTIVE: As an international organization with the missions to promote informatics in health care and biomedical research, advance international cooperation, stimulate research, development and education, and disseminate and exchange information, the International Medical Informatics Association (IMIA) must be constantly cognizant of new developments in medical informatics and address the challenges to the discipline. From an international organization standpoint, it perceives three major challenges viz. the Identity, Organizational and Leadership challenges. METHOD: This paper attempts to identify and discuss these challenges and to offer ways to overcome them through the activities of an international organization for medical informatics. RESULTS AND CONCLUSION: From an international organization standpoint, IMIA can help overcome these organizational challenges by ensuring strong leadership throughout its echelon, actively promoting its goals and objectives worldwide through its national and institutional members as well as its regional groups and encouraging strategic partnerships between its many Working Groups and Special Interest Group on Nursing with other international organizations and industry to further promote the awareness and the perception of the relevance of medical informatics to health and medicine by the international community. PMID- 11933767 TI - Person identification from the EEG using nonlinear signal classification. AB - OBJECTIVES: This paper focusses on the person identification problem based on features extracted from the ElectroEncephaloGram (EEG). A bilinear rather than a purely linear model is fitted on the EEG signal, prompted by the existence of non linear components in the EEG signal--a conjecture already investigated in previous research works. The novelty of the present work lies in the comparison between the linear and the bilinear results, obtained from real field EEG data, aiming towards identification of healthy subjects rather than classification of pathological cases for diagnosis. METHODS: The EEG signal of a, in principle, healthy individual is processed via (non)linear (AR, bilinear) methods and classified by an artificial neural network classifier. RESULTS: Experiments performed on real field data show that utilization of the bilinear model parameters as features improves correct classification scores at the cost of increased complexity and computations. Results are seen to be statistically significant at the 99.5% level of significance, via the chi 2 test for contingency. CONCLUSIONS: The results obtained in the present study further corroborate existing research, which shows evidence that the EEG carries individual-specific information, and that it can be successfully exploited for purposes of person identification and authentication. PMID- 11933768 TI - Design participation as an insurance: risk-management and end-user participation in the development of information systems in healthcare organizations. AB - OBJECTIVES: The aim of this paper is to build a theoretical framework for analysis of when decision-makers should use end-user participation as a form of insurance for unforeseen consequences of implementing information systems in healthcare organizations. METHOD: Data were collected in a case study of an information system development project in a small clinical setting. During the initial phase, the future end-users of the system were allowed to actively influence the system design and test every new tool that was considered for implementation. RESULTS: The results of the case study suggest that when time and effort are invested in allowing healthcare staff to participate in information system development processes, the benefits can well exceed the costs throughout the life cycle of the project. Risk-averse decision-makers fearing negative secondary consequences of a HIS, with regard to clinical work flow, will always adopt measures to prevent future failures, if they can find a possibility of shifting these risks. Therefore, they calculate the present discounted value of the effects accrued over time to the unit and predict the amount of resources they are willing to pay to acquire on insurance (such as design participation) that will protect the organization from future losses. CONCLUSIONS: End-user participation in the design process can be the key positive influence on the quality of the service and, thereby, organizational effectiveness. Investments in broad design participation can, consequently, be a productive activity that transforms potential current income into future benefits. PMID- 11933769 TI - Medical informatics: between science and engineering, between academia and industry. AB - OBJECTIVE: To analyze the nature and appropriate role of the Medical Informatics research and practice area in the 21st Century, and to determine its links to academic environments versus industrial companies and health-care organizations. METHODS: A qualitative analysis of the state of the art of Medical Informatics, based on observation of current medical informatics programs and research in academic and industrial sites. RESULTS AND CONCLUSIONS: Medical Informatics is definitely a scientific and technological area of endeavor, although somewhat ill defined in scope. It is situated between science and engineering, but much closer to the engineering world, and its multidisciplinary nature fits well the engineering paradigm. It is better viewed as a specialization of the informatics field rather than as a basic medical science. However, there are good arguments as to why Medicine should be the first among equals to have its own informatics domain. Medical Informatics must have extensions to both academia and industry to survive. Medical informaticians, whether implicitly or explicitly, exist in three different environments: academic, clinical (user), and industrial (informatics developer); all three environments must be considered when trying to predict the future of this new multidisciplinary area. PMID- 11933770 TI - An Anesthesia Information Management System (AIMS) as a tool for controlling resource management of operating rooms. AB - OBJECTIVES: In our department, we have been using an Anesthesia Information Management System (AIMS) for five years. In this study, we tested to what extent data extracted from the AIMS could be suitable for the supervision and time management of operating rooms. METHODS: From 1995 to 1999, all relevant data from 103,264 anesthetic procedures were routinely recorded online with the automatic anesthesia record keeping system NarkoData. The program is designed to record patient related time data, such as the beginning of anesthesia or surgical procedure, on a graphical anesthesia record sheet. The total number of minutes of surgery and anesthesia for each surgical subspecialty per hour/day and day of the year was calculated for each of the more than 40 ORs, amounting to a total of 112 workstations. RESULTS: It was possible to analyze the usage and the utilization of ORs at the hospital for each day of the year since 1997. In addition, annual and monthly evaluations are made available. It is possible to scrutinize data of OR usage from different points of view: queries on the usage of an individual OR, the usage of ORs on certain days or the usage of ORs by a certain surgical subspecialty may be formulated. These data has been used repeatedly in our hospital for decision making in OR management and planning. CONCLUSIONS: In assessing the results of our study, it should be considered that the system used is not a specialized OR management tool. Despite these restrictions, the system contains data which can be used for an exact and relevant presentation of OR utilization. PMID- 11933771 TI - Industrial action. The same flannel across the channel? Interview by Helene Mulholland. PMID- 11933772 TI - Sexual health. Let's talk about sex. Interview by Matthew Akid. PMID- 11933774 TI - Mental health. Taking the drama out of a crisis. PMID- 11933773 TI - Diabetes. Counting the cost. PMID- 11933775 TI - Should vaccination be compulsory for all preschool children? PMID- 11933777 TI - The NHS is still playing the waiting game. PMID- 11933778 TI - Laughing all the way to the bank. PMID- 11933779 TI - Don't you just love being in control? PMID- 11933781 TI - Improving palliative care. PMID- 11933780 TI - Cancer care put through its paces. PMID- 11933782 TI - Nursing with dignity. Part 4: Christianity II. PMID- 11933783 TI - Pressure ulcer prevention. Part 2. Patient assessment. PMID- 11933784 TI - Cardiopulmonary resuscitation. Mouth-to-mouth ventilation. PMID- 11933785 TI - A specialist asthma service. PMID- 11933786 TI - A nurse-led service for COPD patients. PMID- 11933787 TI - Respiratory physiotherapy. PMID- 11933789 TI - The role of the cystic fibrosis nurse specialist. PMID- 11933791 TI - Examining the role of health services research in public policymaking. AB - Conceptual, methodological, and practical issues await those who seek to understand how to make better use of health services research in developing public policy. Some policies and some policymaking processes may lend themselves particularly well to being informed by research. Different conclusions about the extent to which policymaking is informed by research may arise from different views about what constitutes health services research (is it citable research or any professional social inquiry that can aid in problem solving?) or different views about what constitutes research use (is it explicit uses of research only, or does it also include tacit knowledge or the positions of stakeholders when they are informed by research and are influential in the policymaking process?). Some conditions may favor the use of research in policymaking, like sustained interactions between researchers and policymakers. Results from an exploratory study on the use of health services research by Canadian provincial policymakers illustrate these issues. PMID- 11933792 TI - Extending life: scientific prospects and political obstacles. AB - Aging can be slowed in laboratory rodents by low-calorie diets, and changes in single genes can extend mouse life span by 40 percent or more. Therefore, despite its surface complexity and effects on multiple cells and intercellular systems, aging in mammals might also be retarded by both genetic and nongenetic means. If human aging could be slowed pharmacologically to the extent now possible in rodents, the effect on healthy life expectancy would exceed that of abolishing cancer, cardiovascular disease, and adult-onset diabetes. Why, then, is research on the biological control of aging and longevity poorly funded and shunned by both most scientists and those setting national research priorities? Economic disincentives, disease-specific lobby groups, scientific careerism, and ineffective nostrums, together with gerontologiphobia, must be overcome before such research can improve public health. PMID- 11933793 TI - Not afraid to blame: the neglected role of blame attribution in medical consumerism and some implications for health policy. AB - A crucial aspect of medical consumerism has been overlooked in past research and policymaking: how consumers decide whom to "blame" for bad outcomes. This study explores how, in a system increasingly dominated by managed care, these attributions affect consumers' attitudes and behavior. Using data from the experiences of people with serious mental illness, hypotheses are tested regarding the origins and consequences of blaming for medical consumerism. Blame was allocated to health plans in a manner similar, but not identical, to the way in which blame was allocated to health care professionals. Both allocations are shaped by enrollment in managed care, with blame allocation affecting consumers' subsequent willingness to talk about adverse events. Policy implications include the need for more finely tuned grievance procedures and better consumer education about managed care practices. PMID- 11933794 TI - Pluralism, public choice, and the state in the emerging paradigm in health systems. AB - Today in developed nations, the public pays for most medical care, with the state and the medical profession or providers determining its nature, form, and level. But there is no well-defined institutional framework for revealing consumer preferences and enabling client choice about the nature and form of public entitlement. This thwarts the efforts of health system reformers to satisfy their clients and consequently promote equity and control costs--the raison d'etre of publicly supported care. Consumers can be empowered in the emerging paradigm, however, in which the publicly financed system also contains competing fund holding institutions that organize and manage the consumption of care (OMCC), such as HMOs and sickness funds. In a system in which individuals are entitled to health coverage, OMCC institutions can play an essential role in both shaping the entitlement and in expressing members' preferences. To do this, the OMCCs need to be financed through capitation and endowed with appropriate constitutional rights on how to use the funds. PMID- 11933795 TI - Performance-based contracting in Wisconsin public health: transforming state local relations. AB - In 2000, the Wisconsin Division of Public Health reorganized its allocation of federal and state funds by basing contracts on performance rather than audited costs. This created a quasi market in which the state acted as the buyer and the local health departments as the sellers of public health services. In its first year of operation, the program more effectively defined public health objectives to its funders and constituencies, linked its fiscal accountability more closely to attainment, and documented performance more carefully. In the next two years, the program will focus on improving the quality of objectives and training all parties in negotiation skills. The 2003-6 contract cycle will concentrate on multiyear and multiprogram objectives and a Web-based contract management system. This new contract system will not, however, be established permanently until its long-range impact on funding levels and population health status is known. PMID- 11933796 TI - Nurses needled by mistrust over MMR. PMID- 11933797 TI - History of nursing. Under the white cross. PMID- 11933798 TI - Alcohol misuse. The demon drink. PMID- 11933799 TI - Have you got the perfect body? PMID- 11933800 TI - Should nurses retake control of essential nursing care from HCAs? PMID- 11933803 TI - Spoilt for choice. PMID- 11933804 TI - Good advice given with class. PMID- 11933805 TI - Confessions of a nurse prescriber. PMID- 11933806 TI - The role of practice development in modernising the NHS. PMID- 11933807 TI - Nursing with dignity. Part 3: Christianity I. PMID- 11933809 TI - Pressure ulcer prevention. Part 1. Causes of pressure ulcers. PMID- 11933808 TI - Quality and clinical outcomes in applied relaxation. PMID- 11933810 TI - Cardiopulmonary resuscitation. Applying cricoid pressure. PMID- 11933811 TI - Have skills, will travel. PMID- 11933812 TI - Screening for carcinoma of the colon: current views. PMID- 11933813 TI - Current issues in management of miscarriage and early pregnancy bleeding. PMID- 11933814 TI - Sports nutrition: an overview. AB - Nutritional interventions have the potential to influence the outcome of athletic competition where opponents are closely matched. Sound dietary habits can also influence the adaptations that occur in response to training. This article summarizes some of the strategies that the athlete can use to enhance performance. PMID- 11933815 TI - Eating problems in young children. AB - Over the past 10 years, there has been increasing recognition of the importance of eating problems in young children. Paediatricians are now also recognizing the importance of managing this problem in the context of a wide range of medical disorders. Multidisciplinary assessment and treatment is essential to manage the wide variety of childhood eating problems. PMID- 11933816 TI - Glutamine-enhanced nutrition in the critically ill patient. AB - It is well established that critically ill patients have a deficiency of the amino acid glutamine. This article reviews the evidence for supplemental glutamine in the critically ill, focusing on the benefits in terms of reduced mortality and infectious morbidity. PMID- 11933817 TI - Asbestos-related disease. AB - Until the 1980s, asbestos was widely used throughout the UK. The incidence of asbestos-related disease is still climbing because of the long delay in developing the disease from the initial exposure. The spectrum of diseases encompasses malignant mesothelioma, asbestosis, asbestos-related lung carcinoma and benign pleural disease, including pleural plaques. PMID- 11933818 TI - Diagnosis and management of thyroid eye disease. AB - Recent advances are helping elucidate the pathogenesis and improve the management of thyroid eye disease. While biochemical investigations and imaging may be supportive, ophthalmological and medical clinical assessments remain the key to the diagnosis and management of this sight-threatening disorder. PMID- 11933819 TI - Chloramphenicol in the 21st century. AB - Chloramphenicol use has declined in the UK, despite it remaining highly active against a wide range of bacteria. This article will review the activity, pharmacology, toxicology, uses and potential uses of chloramphenicol in an era of increasing antimicrobial resistance. PMID- 11933821 TI - Anorectal sepsis. AB - Anorectal sepsis is a common cause of hospital admission, presenting with abscess or fistula formation. This article discusses the aetiology of acute anorectal sepsis and its management. PMID- 11933820 TI - Somatuline Autogel: an extended release lanreotide formulation. AB - Somatuline Autogel provides effective control of growth hormone levels and is at least as effective as intramuscular Somatuline LA. Somatuline Autogel has a first order kinetic profile providing at least 28 days' efficacy with reduced burst release and a more consistent drug release over the dose period. The product is presented in a pre-filled syringe and is given deep subcutaneously. PMID- 11933822 TI - Diagnosis and immediate care of soft tissue knee injuries. PMID- 11933823 TI - Pilot study of 360 degrees assessment of personal skills to inform record of in training assessments for senior house officers. AB - The assessment of a trainee doctor's human skills, including teamworking, communication and maintaining trust, is difficult but necessary to confirm competence and to alert trainees and trainers of potential problems before they become intractable. This study used 360 degrees team observation reports for this purpose. The process was easy to administer and valued by trainees. PMID- 11933824 TI - Carcinoma-induced diffuse pulmonary haemorrhage. PMID- 11933825 TI - Bronchoscopic and angiographic findings in Dieulafoy's disease of the bronchus. PMID- 11933826 TI - A builder in a 'staple' condition. PMID- 11933827 TI - Culinary delights in kitchen pathology. PMID- 11933828 TI - Critical letters about writing critical letters. PMID- 11933829 TI - Can we measure the quality of perioperative care? PMID- 11933830 TI - [Metabolic problems and nutrition in burns]. AB - Metabolic changes with burns patients are enormous. The loss of skin substance and the necessity of its reconstruction are at the origin of this exceptional situation. In this context of major aggression with important metabolic alteration the nutritional needs are considerable. To assure a tissue reconstruction, nutrition is as important as the fight against infection. The authors make the point on the quantity needs and the quality needs. They raise up the perspectives concerning immunonutriments and note the importance of enteral administration. As a conclusion, they insist on the fundamental role of the clinical aspect, in the survey of nutritional state. PMID- 11933831 TI - [Local treatment of burns]. AB - Local burn care is one among the components of the total burn care. However since local wounds initiate and maintain the systemic disorders and provoke the aesthetic and functional sequellae, its significance must be stressed. The quality of the results relies upon teams trained and organized to control infection hazards in centres specifically designed. Closure of the wounds, through spontaneous healing of superficial burns or surgical techniques in deep ones, represents the aim of local care which includes the cleaning and dressing of superficial wounds and deep ones before and after grafting, with topical antibacterial (or not) agents, and also excision and grafting procedures. Today skin autografts remain the basic technique for closing deep burns. However because of the inadequacy between donor and wound areas either they must be meshed or other methods are added especially skin allografts, keratinocytes cultures or artificial skin. Local care does not come to an end when wound closure is achieved because of the lengthy rehabilitation cares such as massages, wearing of custom-fitted pressure garments, cures at specialized spas and some patients also need secondary surgery. Burn injuries remain one of the most devastating ordeal that a human being can sustain, because most of them provoke permanent and severe after-effects, whatever their extend and areas, that initial local care must aim at minimizing. PMID- 11933832 TI - [Acute respiratory insufficiency in burn patients from smoke inhalation]. AB - Respiratory injuries by smoke inhalation are one of the most frequent reasons for acute respiratory failure in burn victims. They are most often of chemical origin and are responsible of a 20 to 70% increase of the mortality compared to the mortality of patients with similar burn injuries, but without inhalation lesions. They are often associated to a certain degree to other factors of acute respiratory failure: superior air way obstruction by oedema in face and neck burns, thoracic expansion hindrance due to thoracic burns, lung trauma lesions by blast injury. The generalized inflammatory reaction due to the extent of burns and an initial inadequate resuscitation are worsening factors. The inflammatory process may be responsible of lung injuries similar to those induced by smoke inhalation, even when there is no inhalation. The treatment remains symptomatic and based on the oxygen therapy, mechanical ventilation, prevention of infections and maintain of homeostasis by hydroelectrolytic adequate resuscitation. The nitric oxyde associated to the almitrin allows in a certain number of cases to minimize intra pulmonary shunting and to normalize the VA/O ratio. The development of treatments allowing to modulate inflammatory mediators may lead to news therapies in the future. PMID- 11933833 TI - [The pain from burns]. AB - The painful events associated with the treatment of a severe burn can, because of their long-lasting and repetitive characteristics, be one of the most excruciating experiences in clinical practice. Moreover, burn pain has been shown to be detrimental to burn patients. Although nociception and peripheral hyperalgesia are considered the major causes of burn pain, the study of more hypothetical mechanisms like central hyperalgesia and neuropathic pain may lead to a better understanding of burn pain symptoms and to new therapeutic approaches. Continuous pain and intermittent pain due to therapeutic procedures are two distinct components of burn pain. They have to be evaluated and managed separately. Although continuous pain is by far less severe than intermittent pain, the treatment is, in both cases, essentially pharmacological relying basically on opioids. Because of wide intra- and inter-individual variations, protocols will have to leave large possibilities of adaptation for each case, systematic pain evaluation being mandatory to achieve the best risk/benefit ratio. Surprisingly, the dose of medication decreases only slowly with time, a burn often remaining painful for long periods after healing. Non pharmacological treatments are often useful and sometimes indispensable adjuncts; but their rationale and their feasibility depends entirely on previous optimal pharmacological control of burn pain. Several recent studies show that burn pain management is inadequate in most burn centres. PMID- 11933834 TI - [Psychological aspects of burns]. AB - Destroying the skin, the burn threatens the limits between the self and others. It induces a breakdown in the life which outcome deals with the history of every patient and his psychic structures. But in such a situation, the medical teams have to set up the specific support required by these patients. Through their symptoms the burned patients try to express a deep psychic suffering which needs to be listened to. PMID- 11933835 TI - [Burns and rehabilitation]. AB - Necessary principles of rehabilitation for burn patient are based on empirical findings recently corroborated by discoveries about healing pathophysiology. Risks are assessable immediately from the extensive, depth and situation of the burns, problems appear only if the dermis is affected: retraction, hypertrophy and losses of substances. To cutaneous problems it is necessary to add those linked to the prolonged immobilization and to complications of the resuscitation. To be effective, re-education has to be precocious, continuously suited to cicatricial processing and to the different therapeutic steps: resuscitation, surgical treatment, processing in a re-education and rehabilitation center, steady at home and processing of the sequelae. The processing rests on the repressive cloth port 23/24 hours during more of a year, the port of orthesis of immobilization and segmental posture (to stretch the dermis permanently) and the mobilization of articulations to avoid their stiffening. The cooperation of the patient is essential, it needs the share of therapies as well as the totality of problems and difficulties met by the patient, that they are physical, psychological, social, family or occupational. The steady has to be insured by a pluridisciplinarity team during at least the two necessary years for the cicatricial maturation. PMID- 11933836 TI - [Criteria for burn severity. Epidemiology. prevention, organization of management]. AB - Total body surface area burned and deep of the skin injury are the main determinants of the burn severity. Other factors like age of the patient, pathological conditions, pulmonary injury by smoke inhalation, wound localizations play also a major role. 500,000 cases of burn injuries occurred each year in France. Ten hundred are hospitalized among witch 3,000 are hospitalized in burn units. Burns by flames are the most common in adult severely burned patients. In children, for all type of injuries, and in adult patients suffering of light or medium injuries, hot liquids are the most frequent encountered agents. Obviously, prevention programs should markedly decrease the occurrence of burn injuries. Regulation modifications are probably more potent than information campaigns. There is 25 burn units in France. Unfortunately, these units take care of less than one third of all the burned hospitalized patients. The organization of a national network binding regional burn centers and local specialized units would probably improve the efficiency of burn therapy in our country. PMID- 11933837 TI - [Prehospital management of burns]. AB - The optimal assessment of the burn victim in the prehospital setting needs a clear understanding of the pathophysiological changes occurring locally and systemically after injury. For the mobile emergency unit it is essential to dispose of an appropriate protocol for the prevention of hypovolemia, hypothermia, hypoxemia. In the immediate survey, immediate life-threatening conditions should be clearly identified and treated as well as a through head-to toe evaluation should be undertaken. Two phases of transport can be identified: the first from the scene of accident to the proximal emergency unit, specialized or not in burn care. The second is the transportation from the non-specialized proximal emergency unit to a burn intensive care unit according to the decision of the dispatching Centre 15. PMID- 11933838 TI - [Initial shock from burns. Physiopathology: therapeutic principles]. AB - Widespread destruction of the skin induces a large necrotic mass and a break of the skin barrier. It also leads to an intense inflammatory reaction. This activates keratinocytes, endothelial cells and neutrophils. Certain mediators (e.g. endothelin, histamine, bradykinin, serotonin, catecholamines, vasopressin, prostaglandins, cytokines and nitrogen monoxide) are thus released in large quantities and act both at the site of the burns and at a distance. The abnormally high level of albumin in the capillary wall and the increased capacity of absorption of the interstitial areas around the burns are the main abnormalities observed. This results in a hypovolemia associated with a hemoconcentration, hyponatremia, hypoalbuminemia, systemic vasoconstriction and myocardial malfunction, which is difficult to evidence. During the initial phase, the major risk is the appearance of hypovolemic shock, which is rapidly irreversible if early treatment is not administered. Vascular filling with iso- or hyper-osmolar sodium crystalloids, associated with buffer solutions, is the first line. There is still debate regarding the best moment at which to give albumin. A hyperkinetic shock may occur after several hours and despite the filling. The symptoms are tachycardia, increased heart rate and a dramatic decrease in systemic vascular resistance. This may lead to metabolic acidosis and multi-organ failure. Study of the hemodynamic profile of the patient allows the rational use of pressor amines and haemodialysis. PMID- 11933840 TI - Synthesis and antiproliferative activity in vitro of new propargyl thioquinolines. AB - The series of new 3,4-disubstituted thioquinolines which possess one or two O, S, Se-propargyl groups has been synthesized on the basis of the reaction of thioquinanthrene with alkoxides. All the compounds obtained were tested for their antiproliferative activity in vitro against the cells of three human cancer cell lines: SW707 (colon cancer), T47D (breast cancer), and HCV29T (bladder cancer). Two compounds, 4-(3-hydroxypropoxy)-3'-propargylthio-3,4'-diquinolinyl sulfide (3) and 3-methylthio-4-propargylselenoquinoline (13) exhibited significant cytostatic activity (ID50 < 4 micrograms/ml) against the cells of all the human cancer lines used and are good candidates for further anticancer activity studies in vitro using a broad panel of human and murine cell lines and for in vivo preclinical screening in different mouse transplantable tumor models. PMID- 11933839 TI - [Inflammatory reaction and infection in severe burns]. AB - Major burn injury is a lesion where the inflammatory reaction is exported to the whole body. After a short time of hemodynamic changes, this inflammation is kept by necrotic tissues, persistence of an opened wound, and by the pulmonary and gut reactions. When infection starts, it becomes difficult to distinguish its symptoms among the inflammatory signals. The main point of the care of burn patient consists in trying to control this reaction and the immuno-depression it leads to: early excision and grafts, early enteral nutrition, perfect nursing care. There is no specific medical treatment of this state. The antibiotic use must be well weighed up. Infection is often the trigger of the multiple organ dysfunction which is the way the burn patient dies but is not mandatory. PMID- 11933841 TI - Synthesis of some new antianemics I. Iron pullulan complexes of pharmaceutical interest. AB - First experiments were performed in order to obtain a polynuclear iron(III) complex with pullulan, which could be applied in therapy of sideropenic anemia. Complete synthesis of the complex can be realized with oligomers of depolymerized pullulan at an average molar mass 8000-10,000 g.mol-1. Maximum iron contents of 51.4 mg.cm-3 is effected by pullulan depolymerizates (Mw approximately 9000 g.mol 1, [eta] = 0.085 dl.g-1) at 130 degrees C for 240 min, at pH = 10.5 and at an initial mass ratio Fe/ligand of 1:3. The effected concentrations and complex stability respond to requirements of pharmacological application of parenteral antianemic preparations. PMID- 11933842 TI - [Furo(2,3-c)quinolines and furo(3,4-c)quinolones--synthesis and investigation of lipoxygenase inhibition]. PMID- 11933843 TI - Direct enantiomeric resolution of some cardiovascular agents using synthetic polymers imprinted with (-)-S-timolol as chiral stationary phase by thin layer chromatography. AB - Molecular imprinted polymers (MIPs) of S-timolol were prepared as chiral stationary phases (CSPs) in thin layer chromatography (TLC). The resolution of the enantiomers of some cardiovascular drugs, including propranolol, atenolol, timolol, nadolol, nifedipine and verapamil were investigated on these CSPs. A mobile phase system of either methanol or acetonitrile was used and the effects of acetic acid content of the mobile phases were also investigated. The best resolution was achieved for enantioseparation of propranolol, timolol and atenolol on plates based on MIP of (-)-S-timolol using methacrylic acid as functional monomer (alpha = 1.52, 1.6, 1.59) respectively, using acetonitrile containing 5% acetic acid and (alpha = 1.47, 1.52, 1.5) in methanol containing 1% acetic acid as mobile phases. The results obtained show that TLC based on MIPs could be applied in the direct separation of several beta-adrenergic drugs. As the side chains on beta-blockers are similar, it is possible that this method could also be used for the resolution of other racemates in this family of drugs. Racemic drugs structurally related to print molecule, were completely resolved into two spots with the MIP plates. In general the retention of (+)-R-isomers was greater than that of (-)-S-isomers, indicating lower stereoselectivity of the MIPs to the dextrorotatory isomers. The method offers a rapid, sensitive and reliable method for quality control for these drugs. PMID- 11933844 TI - Colour reactions of PH. EUR. for identification of drugs using 1,3-dibromo-5,5 dimethylhydantoin (DBH) instead of elemental bromine. Analytical methods of pharmacopoeias with DBH in respect to environmental and economical concern, Part 10. AB - PH. EUR. 2002 and supplements identify aloes (Rosenthaler reaction), amiloride hydrochloride, chlorhexidine, dienestrol, quinidine sulphate, quinine hydrochloride and quinine sulphate (Thalleioquine reaction) and trifluoperazine hydrochloride using elemental bromine. This colour reaction can be performed better with 1,3-dibromo-5,5-dimethylhydantoin (DBH). Some prescriptions of PH. EUR. have been improved in respect to environmental and economical concern. The identification of amiloride hydrochloride with bromine water according to PH. EUR. 2002 or with DBH shows no UV fluorescence as reported in the pharmacopoeia. PMID- 11933845 TI - A simple method for the identification and assay of extracellular plant beta galactosidase. AB - A simple, rapid and reproducible procedure for the identification of extracellular Californian poppy (Eschscholzia californica Cham.) beta galactosidase is described using callus cultures of seedlings from the tested plant, roots of 4-days-old seedlings of Californian poppy germinating on agar plates and cell suspension cultures cultivated from callus cultures. 6-Bromo-2 naphthyl-beta-D-galactopyranoside and p-nitrophenyl-beta-D-galactopyranoside were used as substrates for the determination of the intracellular and extracellular activities of beta-galactosidase. The extracellular beta-galactosidase activity was identified by evaluating the dye-zones in an agar medium. The enzyme from Californian poppy callus cultures or from seedling roots cultivated on agar plates supplemented with 6-bromo-2-naphthyl-galactopyranoside hydrolyzed this substrate releasing 6-bromo-2-naphthol. By simultaneous coupling with hexazonium p-rosaniline the corresponding (reddish-brown) azo-dye was formed. The agar plate method described permits rapid, simple and specific detection of plant producers of extracellular beta-galactosidase. PMID- 11933846 TI - The separation of hypericine and pseudohypericine from Hypericum perforatum L. AB - The extraction of Hypericum perforatum L. was carried out by the method of ultrasonic maceration. The separation of H. perforatum L. extract in order to obtain hypericine and pseudohypericine rich fractions was carried out by the following chromatographic methods: flash column chromatography, high speed countercurrent chromatography, XAD solid phase extraction and Sephadex column chromatography. The separation by the Sephadex column chromatography gave the best results. Preparative HPLC was used to isolate hypericine and pseudohypericine from fraction 4 obtained by the Sephadex column chromatography procedure. PMID- 11933847 TI - Development of a rheometric technique to measure the mucoadhesive capacity of a dry powder formulation. AB - In the present study, the use of an in vitro technique applying oscillatory shear rheology to determine the interaction between a dry polymer powder and mucin is investigated. The polymers examined are Carbopol 974P, Noveon AA1 and a polymer mixture containing Carbopol 974P and Drum Dried Waxy Maize Starch. The degree of interaction is assessed by analysing the variation of the elastic and viscous modulus as a function of oscillation stress. Both moduli are used to calculate a mucoadhesive index at a selected single stress/frequency combination. The contribution of physical entanglements and secondary bond formation to the interaction can be derived from the mathematical analysis of the relation between the moduli and the oscillation frequency. When considering the stress sweep data, the interaction between mucin and the various polymers investigated seems very similar. This similarity is also observed when calculating the mucoadhesive indexes. On the contrary, the mathematical analysis of the frequency sweep data clearly indicates that the secondary bond formation between the polymer powder particles and mucin can be obtained only in the case of Carbopol 974 P. PMID- 11933848 TI - Pluronic F-127 gels as a vehicle for topical formulations of indomethacin and rheological behaviour of these formulations. AB - Topical gel formulations containing a non-steroidal anti-inflammatory drug, indomethacin (IND), were prepared using 20% w/w Lutrol PF-127 as a gel-forming agent, and 16, 20 and 24% w/w Hexylene glycol (HG) or polyethylene glycol 300 (PEG) as solvents. 1% w/w Tween 80 and 1% w/w PVP 25 were added as excipients. The effects of the amounts of solvent and excipients on the physical characteristics of IND gel such as consistency, appearance, crystallization, pH and viscosity were studied. The results indicated that 1% w/w IND is able to form a structural gel. The viscosity values were calculated from the rheograms which were determined by a Haake Rotovisco sensor at a shear rate of 10,000 l/s. Viscosities corresponding to shear rates of 1000, 3000, 6000 and 9000 l/s were also calculated. Yield points were approximated from the rheograms. Although all IND gels maintained their pseudoplastic flow behaviour, their viscosities decreased markedly with increasing shear rates. Furthermore, increasing the amount of HG or PEG gave a more viscous gel except for the 24 w/w% HG gels which turned a jelly with or without either Tween or PVP. The difference in viscosities was explained by the changes in the gel compositions. 20% of PEG-1% PVP ranked first in viscosity followed by 16% PEG-1% PVP, 16% PEG-1% Tween, 24% PEG, 20% PEG 1% Tween and 16% HG-1% PVP. The results indicate that the excipients influence the physical characteristics of the gels. The optimum concentration for gels manifesting as strength of gel was 20% PEG in combination with 1% PVP which had the highest viscosity and yield value at a low shear rate. PMID- 11933850 TI - New indolocarbazoles as antitumour active compounds: evaluation of the target by experimental and theoretical studies. AB - A series of indolocarbazoles was synthesized as congeners of the natural lead compounds rebeccamycin (1) and staurosporine (2) which reduce cell growth by inhibiting topoisomerase I and protein kinase C respectively. Two of the carbazoles (3 and 4) screened at the National Cancer Institute (NCI, USA) showed an interesting cytotoxic activity and were therefore further analysed. The mechanism of action of these two compounds was studied experimentally using different assay to determine the B-DNA binding ability and the inhibition of topoisomerase I and of protein kinase C. Theoretical molecular modelling studies were also performed to describe the possible interactions with protein kinase C and DNA. PMID- 11933849 TI - Studies on solubility and hypoglycemic activity of gliclazide beta-cyclodextrin hydroxypropylmethylcellulose complexes. AB - This study was undertaken with an objective to increase the dissolution rate and bioavailability of a poorly water soluble drug gliclazide (Gz) by complexation with beta-cyclodextrin (CD) in the presence of hydroxypropylmethylcellulose (HPMC). Phase solubility studies of Gz were performed in aqueous solutions of different concentrations of CD alone and in the presence of some water soluble polymers. Gz-CD complexes were prepared in 1:1 and 1:2 drug:CD molar ratios by autoclaving, neutralization and kneading methods. The complexes were also prepared in the presence of 0.05% w/w HPMC. Physical mixtures of Gz-CD in 1:1 and 1:2 molar ratios were also prepared. Complexes and physical mixtures were characterized and evaluated for in vitro dissolution in distilled water and hypoglycemic activity in rats. CD enhanced the dissolution of Gz to 1.5 to 2.0 fold. Presence of water soluble polymer HPMC in Gz-CD complexes further enhanced the rate and extent of drug dissolution to 2.5 fold. Gz-CD-HPMC complexes were found to be more promising as they produced not only an early onset but also more intense hypoglycemic effect as compared to pure drug powder and commercial tablets. PMID- 11933851 TI - Evaluation of toxic activity of 2,4-dihydroxythiobenzanilides. AB - 2,4-Dihydroxythiobenzanilides represent a new group of compounds with significant fungistatic and bacteriostatic properties. The results of investigations on their cytotoxicity are also very convincing. Therefore LD50 doses were determined for five compounds, they ranged from 239 to 840.5 mg/kg. The results of the tests for spontaneous locomotor activity and hexabarbiturane sleeping time indicate low toxicity of the compounds tested. PMID- 11933852 TI - Main constituents of a commercial Drosera fluid extract and their antagonist activity at muscarinic M3 receptors in guinea-pig ileum. AB - The range of known constituents of Drosera species is extended by identification of myricetin 3-O-galactoside, from D. madagascariensis, and (+)-cis isoshinanolone, obtained from a commercial fluid extract. They are accompanied by the naphthoquinones droserone and plumbagin, typical of this taxon, and a series of ubiquitous flavonols, including the rarely found gossypitrin present in the latter source. Conspicuously, the commercial form of D. peltata, non-accepted by the commission E, was found to be devoid of flavonoids. In addition, the fortuitous availability of the authentic enigmatic sample 'CON', previously isolated from D. rotundifolia, led to its characterization as common quercetin. Experiments performed on isolated guinea-pig ileum demonstrated that quercetin respectively 'CON' moderately inhibited carbachol-induced contractions at 10 microM (pD'2 5.09 +/- 0.02), while (+)-cis-isoshinanolone (100 microM) was inactive. This result indicates that quercetin derivatives may well contribute to the therapeutic use of Drosera preparations. PMID- 11933853 TI - Gastroprotective and antiulcerogenic effects of Rumex patientia L. extract. AB - Gastroprotective and antiulcerogenic effects of the aqueous extract of Rumex patientia L. roots (D-1) were investigated in rats and compared to non-steroidal antiinflammatory drugs (NSAIDs). Whereas oral administration of high doses of NSAIDs caused stomach damage, D-1 (aqueous extract of Rumex patientia roots, 150 and 500 mg/kg) and nimesulid (100 and 300 mg/kg) did not cause any damage. In rats, the number of stress ulcers induced by the administration of 150 mg/kg of D 1 were identical to that of the control group. However, compared to the control group, the number of stress ulcers decreased 1.9 times and the size of ulceration areas decreased 1.1 times in the group treated with 500 mg/kg of D-1. PMID- 11933854 TI - Two new chlorinated amides from Nicotiana glauca R. Graham. AB - Two new chlorinated amides, N-(2',6'-diethyl phenyl)-2-chloroacetamide (1) and N (butyloxymethyl)-N-(2',6'-diethyl phenyl)-2-chloroacetamide (2) were isolated for the first time from the ethanolic extract of the leaves of Nicotiana glauca R. Graham in addition to triacontanol (3), scopoletin (4) and stigmasterol-3-beta-O D-gluco-pyranoside (5). The structures of the isolated compounds were elucidated by spectroscopic analysis (1D, 2D NMR, EIMS, HR-EIMS, IR and UV). PMID- 11933855 TI - Two new steroids from Adenophora stenanthina subsp. xifengensis. AB - Two new steroids, 3 beta-vanilloyloxy-stigmast-5-ene-7-one, and stigmast-5-ene-3 beta,7 beta,15 alpha-triol were isolated from the roots of Adenophora stenanthina subsp. xifengensis along with four known compounds, stigmast-5-ene-3 beta-ol-7 one, stigmast-5-ene-3 beta,7 beta-diol, syringin, sinapyl alcohol 1, 3'-di-O-beta D-glucopyranoside. Their structures were elucidated on the basis of spectral methods. PMID- 11933856 TI - Tautomerism between exomethylene type physalins and oxymethylene-bridged physalins. PMID- 11933857 TI - Potassium titanyloxalate as analytical reagent for micro-quantitative determination of quercetin. PMID- 11933858 TI - Inhibition of 3 alpha-hydroxysteroid dehydrogenase by polyene macrolides. PMID- 11933859 TI - [Predictive gene tests find no applications by life and health insurance carriers]. PMID- 11933860 TI - [Nonalcoholic fatty liver]. AB - Nonalcoholic fatty liver disease (NAFLD) is most often associated with obesity, type II Diabetes mellitus, hyperlipidemia and chronic viral hepatitis C. The spectrum of changes encompasses fatty liver, steatohepatitis, liver fibrosis and cirrhosis. Most patients are asymptomatic. The aminotransferases are only slightly elevated (ALT > AST). Grade of inflammation and stage of fibrosis can be assessed accurately only by histologic examination of liver biopsy. In most cases prognosis is favourable but in a subgroup of patients NAFLD may progress to cirrhosis. Recent data suggest that up to 70% of cryptogenic cirrhoses are accounted for by nonalcoholic steatohepatitis. At the moment therapeutic modalities of proven value are not available. PMID- 11933861 TI - [Extraintestinal manifestations in chronic inflammatory bowel diseases]. AB - 60-80% of patients with inflammatory bowel diseases (IBD) develop extraintestinal manifestations. These complications are mostly the result of the underlying disease, deficiencies, or drug therapy and can significantly reduce the quality of life of patients with IBD. The most evident manifestations affect the perianal region, joints, skin, and eyes. Regular check-ups and early substitution in addition to a consistent therapy of the chronic intestinal inflammation usually lead to a significant reduction of the extraintestinal symptoms. PMID- 11933862 TI - [Freiburg intervention program for ambulatory therapy of obesity in childhood (FITOC)]. AB - In this study we first try to answer the question, whether it is possible to make a successful treatment for obese children in an interdisciplinary program. Second it is asked whether a transfer of this program to further regions in Germany leads to comparable results. In FITOC children from the age of 8-11 years and over the 97. BMI-percentile are integrated in this program. The goals weight management, increased physical fitness and improvement of the cardiac risk profile are checked by weight, height, fasting blood serum, a standardized cycle ergometry and a medical measurement at the beginning, after treatment and at all check-ups. The recorded medical data show clearly that the intervention leads to a significant improvement in almost all checked parts. The successful treatment can be recorded after 8 months, likewise after 2.5 years as a long-term result. The further cornerstones of FITOC nutrition and psychology are not subject of this publication. In future the psychological part in FITOC will be evaluated by standardized inventories. The group from Duren has a success in therapy according to the definition of the program. Thereby it is shown that FITOC is extendable, if teams are trained intensively and the conditions are comparable. FITOC is able to treat obese children successfully over a long period of time. In consideration of the rising prevalence of obesity in childhood and the limited financial resources in health care this outpatient interdisciplinary program is an effective choice of treatment. PMID- 11933863 TI - [Socioeconomic significance of dementia and mild cognitive disorders]. AB - While diagnostic criteria and epidemiologic results are available for dementia, reliable quantitative information on mild cognitive impairment is rare. The distinction between mild cognitive impairment, effects of normal ageing and dementia can be difficult to establish. Conceptual problems notwithstanding, a growing number of patients with dementia and with mild cognitive impairment can be safely predicted. Reduced quality of life on the part of patients and their families, growing demand for care and increasing costs of illness will prove the socioeconomic relevance of dementia. PMID- 11933864 TI - [Cardiomyopathies]. AB - Cardiomyopathies are characterized by ventricular enlargement, thickness or stiffness. According to the WHO classification, diseases of unknown origin (cardiomyopathies) are distinguished from heart muscle diseases with known origin or having a relationship with other organ systems (specific heart muscle diseases). Cardiomyopathies are characterized by a worsening of cardiac work resulting from failure of the pump function and ventricular compliance or from severe arrhythmias. Consequences in prognosis and work capacity depend on the degree of heart failure and arrhythmias. PMID- 11933865 TI - [Cultural historical significance of gout. A portrait of German social history in the 20th century]. PMID- 11933866 TI - [Comment on C. Sieger: High life expectancy--a risk?]. PMID- 11933867 TI - [Comment on R. Lehmann: Evaluation of erectile dysfunction in private accident insurance]. PMID- 11933868 TI - Rats kick their drinking habits. PMID- 11933869 TI - Proceedings of the American Veterinary Medical Association's Eleventh Annual Animal Welfare Forum. November 10, 2000. Itasca, Illinois, USA. PMID- 11933870 TI - [An outstanding organizer of military forensic-medical expertise (on the 100th anniversary of the birth of M.I. Avdeev)]. PMID- 11933871 TI - [Mikhail Vladimirovich Portnoi (on the 80th anniversary of his birth)]. PMID- 11933872 TI - [A prominent military epidemiologist (On the 70th anniversary of the birth of V. S. Perepelkin)]. PMID- 11933873 TI - Late breaking abstracts for the 31st European Society for Dermatological Research (ESDR) Meeting 2001. September 20-21, 2001. Stockholm, Sweden. PMID- 11933874 TI - Proceedings of the 33rd Congress of the Japan Epilepsy Society. October 22-23, 1999. Sendai, Japan. PMID- 11933875 TI - Program and Abstracts of the 23rd Annual Winter Neuropeptide Conference. February 2-5, 2002. Breckenridge, Colorado, USA. PMID- 11933876 TI - [Fall congress NVT: 'Human-work']. PMID- 11933877 TI - [Oro-antral perforations. Desirability of antibiotic support in surgical closure within 24 hours]. AB - OBJECTIVE: The routine use of antibiotics in oro-antral perforations is advocated by a number of authors, in order to prevent post-operative complications such as sinusitis. Our aim was to establish wether the preventive use of antibiotics seams to be indicated in the closure of this perforations. DESIGN: A retrospective investigation. SETTING: The department of Oral and Maxillofacial Surgery and Oral Pathology of the Academic Center for Dentistry Amsterdam (ACTA) and the Free University Hospital Amsterdam. METHODS: In a retrospective study 46 patients have been investigated, 32 males and 14 females, who underwent a surgical closure of an oro-antral perforation in 1995, within 24 hours after its occurrence, in a department of oral and maxillofacial surgery. Twenty-two patients received preoperative antibiotics and 24 patients were treated without the use of antibiotics. RESULTS: Three (6.5%) patients had a post-operative sinusitis. In the other 43 patients there were no postoperative complications. CONCLUSION: The preventive use of antibiotics in oro-antral perforations in order to prevent post-operative complications such as sinusitis seams not to be indicated in the closure of this perforations. However, before a final conclusion on this matter can be drawn, a prospective randomized study should be undertaken. PMID- 11933878 TI - Steroid therapy of acute ENT infections: rarely indicated. AB - (1) Apart from acute laryngitis in children, the use of steroids in acute ENT infections is not supported by clinical data. (2) A single dose of steroids (oral or intramuscular dexamethasone, 0.6 mg/kg) has only moderate efficacy in children with acute laryngitis, but it can hasten symptom relief. Available clinical data fail to show whether steroid therapy reduces the frequency of severe respiratory complications in this setting, or if it is helpful in minor cases. (3) There are no published data justifying the use of steroids as adjuvant treatment in other acute ENT infections, such as non allergic rhinitis, sinusitis, pharyngitis, tonsillitis and otitis. Two randomised trials have shown an analgesic effect of steroids in pharyngitis, but there are no published comparisons with standard analgesics such as paracetamol. (4) Severe complications appear to be rare with single-dose and short-term steroid therapy (for less than a week). However, there is a potential risk of rare but severe complications of chickenpox, and avascular necrosis of the femoral head. (5) Routine use of steroids for recurrent ENT infections has the same risks as long-term steroid therapy. PMID- 11933880 TI - Selected abstracts from presentations to the 18th Autumn Meeting of the Association of Cardiothoracic Anaesthetists. November 16, 2000. London, UK. PMID- 11933879 TI - Abstracts for the Paleoanthropology Society Meeting. March 19-20, 2002. Denver, Colorado, USA. PMID- 11933881 TI - Contributions of the Third Symposium of the Donald B. Brown Research Chain on Obesity. November, 17-18, 2000. Quebec, Canada. PMID- 11933882 TI - Are you preparing for HIPAA? PMID- 11933883 TI - Infectious agents as biological weapons. PMID- 11933884 TI - Instrumental changes. New certification helps to ensure quality. PMID- 11933885 TI - HSJ management challenge. Call of the wild. PMID- 11933886 TI - Proceedings of the Second Congress of Asia Pacific Symposium on Cochlear Implant and Related Sciences, April 2-4, 1999. Seoul, Korea. PMID- 11933887 TI - Development of an algorithm for improving the description of the pulmonary pressure-volume curve. PMID- 11933888 TI - Pulmonary nitric oxide uptake reflects the entire diffusive properties of the alveolar capillary membrane. PMID- 11933889 TI - ACE genotype and performance. PMID- 11933890 TI - Genetic studies of performance. PMID- 11933891 TI - Genetic studies of performance. PMID- 11933892 TI - Image of the month. Mantle cell lymphoma. PMID- 11933893 TI - Smallpox - a potential biowarfare weapon: destroy the virus or keep the vaccine for ever. PMID- 11933894 TI - Analysis of digital scanning laser ophthalmoscopy fundus autofluorescence images of geographic atrophy in advanced age-related macular degeneration. AB - BACKGROUND: Fundus autofluorescence (AF) imaging using confocal scanning laser ophthalmoscopy (cSLO) has been shown to be superior to fundus photography or angiography for delineating areas of geographic atrophy (GA) in retinal pigment epithelium (RPE) and for recording variation over time. We have evaluated a method for automated computerized detection and quantitation of RPE atrophy. METHODS: AF images in vivo were recorded with a confocal scanning laser ophthalmoscope (exc. 488 nm, em. >500 nm; Heidelberg Retina Angiograph). The intensity of AF in atrophic areas was markedly decreased. Two independent readers analysed these areas in 24 right eyes manually by outlining GA areas using a mouse-driven arrow (method A) and automatically by image analysis software (Global Lab Image/2) after subjective adjustment of thresholding (method B). Agreement between observers and between methods A and B was assessed by the Bland Altman design for method-comparison studies. RESULTS: Larger areas were measured using method A than B by both readers (agreement A/B: reader 1 mean difference 1.04 mm, 95% CI [0.66,1.42]; reader 2 mean difference 0.62 mm, 95% CI [0.43,0.81]). The agreement between the readers was mean difference 0.39 mm (95% CI [0.02,0.76]) for A and mean difference -0.03 mm (95% CI [-0.23,0.18]) for B. Features making the delineation of borders of GA difficult included large choroidal vessels with autofluorescent properties in the GA area and media opacities. CONCLUSIONS: Fundus AF cSLO imaging provides a reliable means to delineate areas of GA. The automated image analysis allows more accurate detection and quantitative documentation of atrophic areas than manual outlining. This method will be useful in longitudinal natural history studies and for monitoring effects of future therapeutic interventions to slow down GA progression in patients with advanced atrophic ARMD and other retinal diseases associated with outer retinal atrophy. PMID- 11933896 TI - Oral tumours in Tanzanian children based on biopsy materials examined over a 15 year period from 1982 to 1997. AB - AIM: To provide information on the types and distribution of oral tumours and tumour-like lesions occurring in a Tanzanian child population aged 0-16 years. DESIGN: Retrospective study of biopsy results from hospital records from 1982 1997. SETTING: Department of Histopathology, the Muhimbili Medical Centre (MMC) in Dar es Salaam. INTERVENTION/ METHODS: A total of 158 biopsy results, from 75 girls and 83 boys, were retrieved and studied. RESULTS: Malignant tumours were the most frequent (43.0%) followed by benign tumours (30.4%) and tumour-like lesions (26.6%). Burkitt's lymphoma was the most frequent malignant tumour accounting for 88.2% of all malignancies followed by squamous cell carcinoma (4.4%) and oral Kaposi's sarcoma (2.9%). Fibroma, papilloma and haemangioma were the most frequent benign tumours constituting 27.1%, 14.6% and 12.5%, respectively. Odontogenic cysts were the most frequent tumour-like lesions (28.6%) followed by fibrous dysplasia (19%) and giant cell granuloma (16.7%). CONCLUSION: The six most common oral lesions were Burkitt's lymphoma, fibroma, odontogenic cysts, fibrous dysplasia, papilloma and giant cell granuloma. PMID- 11933895 TI - Aging effects on memory encoding in the frontal lobes. AB - Functional magnetic resonance imaging (fMRI) was used to compare frontal-lobe activation in younger and older adults during encoding of words into memory. Participants made semantic or nonsemantic judgments about words. Younger adults exhibited greater activation for semantic relative to nonsemantic judgments in several regions, with the largest activation in the left inferior frontal gyrus. Older adults exhibited greater activation for semantic judgments in the same regions. but the extent of activation was reduced in left prefrontal regions. In older adults, there was a significant association between behavioral tests of declarative and working memory and extent of frontal activation. These results suggest that age-associated decreases in memory ability may be due to decreased frontal-lobe contributions to the initial encoding of experience. PMID- 11933897 TI - Radiographic prevalence of approximal enamel lesions and relationship with dentine lesions and restorations in Dutch adolescents. AB - AIM: The aim of this study was to assess the prevalence of approximal enamel lesions from radiographs in groups of 14, 17, 20 and 23 years old, and to determine the relation with dentine lesions and restorations present. METHOD: Per age group, the bitewings of 120 persons were randomly selected and assessed for enamel lesions from the distal surface of the first premolar to the mesial surface of the second molar. A second examiner assessed 20% of the material for enamel lesions to determine the interexaminer agreement (Cohen's kappa = 0.63). RESULTS: An enamel lesion was found in 12-15% of the unfilled surfaces. The mean number of surfaces with enamel lesions was almost three per person. For all age groups it was found that about 20% of the persons had at least four enamel lesions and the number of enamel lesions was significantly correlated with the number of dentine lesions. Of all enamel lesions 20% were found in persons without any dentinal lesions or restorations. CONCLUSION: The bitewing radiographs showed a considerable number of enamel lesions for these age groups. This seems to justify its use on a routine base as a diagnostic tool at the age of 14-16 years. PMID- 11933898 TI - Risk indicators for tooth wear in New Zealand school children. AB - AIM: To investigate the prevalence and severity of tooth wear in the primary dentition of a representative sample of New Zealand school children and relate these to possible risk factors. DESIGN: A cross-sectional study. SETTING: Primary schools in Dunedin, New Zealand. PARTICIPANTS: 104 children of both sexes, aged between 5 and 8 years, randomly selected. METHODS: Clinical examinations of the buccal, occlusal/incisal and lingual surfaces of deciduous canines and molars. MAIN OUTCOME MEASURE: Degree of wear and the presence of dentinal cupping of teeth. Information on weaning and consumption of fruit-based drinks at bed time, frequency of consumption of fruits, yoghurt, pickled foods, fizzy and fruit-based drinks. RESULTS: The prevalence of tooth wear was similar in boys and girls and there were no significant differences between sides of the arches. A high percentage (82%) of children had at least one primary tooth with dentine exposed. While maxillary canines showed the greatest prevalence of dentine exposed, maxillary molars displayed the greatest prevalence of cupping. Severe tooth wear was less prevalent among children weaned after 12 months (14.3%) than those weaned earlier (27.9% P < 0.01). There were no statistically significant associations between wear and the consumption of fruit, yoghurt, pickled foods, fizzy drinks or fruit-based drinks. CONCLUSIONS: Tooth wear associated with dentine exposure is common in 5-8 year old children. This is not significantly associated with dietary factors, but appears to be related to early weaning from the breast. PMID- 11933899 TI - Scrub typhus: a rare cause of multiorgan failure in a surgical patient presenting with deep venous thrombosis. PMID- 11933900 TI - Lichen planus following hepatitis B vaccination in an African girl. AB - Lichen planus is a papulosquamous disorder with distinctive clinical features. Until now the aetiology has been largely unknown, however, several postulations have been proposed. The immunopathogenic basis has been most favoured. We report a case of a 13-year-old girl who presented with Lichen planus after receiving the second dose of hepatitis B virus (HBV) vaccination. This report, similar to earlier reports, appears to support the immunopathogenic basis of Lichen planus. PMID- 11933902 TI - Creating and using a digital image library: reasons and methods. PMID- 11933901 TI - Corrosive oesophagitis in Nigeria: clinical spectrums and implications. AB - Ingested corrosive agents produce oropharyngeal and gastro-oesophageal injuries ranging from minor burns to severe necrosis, depending on the agent, amount, concentration and duration of exposure. A 10-year (1990-1999) study of 23 corrosive oesophagitis patients depicting the clinical spectrums and implications was done. There were 17 (74%) males and 6 (26%) females, with an average age of 16 years: 35% were in the first decade of life and 39% in the second, with an average occurrence of three new cases a year. The main corrosive agents were alkali (NaOH) 48% and sulphuric acid 39%. The factors that predisposed to corrosive ingestion included: accidental 43%; suicidal 35%; homicidal 13% and 'therapy' 9%. Of the accidental cases 60% were seen in children; 75% of suicide attempts were in the second decade of life. Eleven (48%) patients developed complications, namely laryngeal stenosis 4% and oesophageal strictures 44%. The hospital stay ranged from 2-4 weeks. Awareness of the dangers of corrosive ingestion should be extensively publicized. Government regulations permitting the sales and procurement of corrosive substances should be strictly enforced and, if not yet in place, should be enacted without further delay. PMID- 11933903 TI - Effects of in-office tooth whiteners on hardness of tooth-colored restoratives. AB - This study investigated the effects of in-office tooth whiteners on the hardness of hybrid (Spectrum TPH), polyacid-modified (Dyract AP), PRG (Reactmer) composites and a resin-modified glass ionomer cement (Fuji II LC). Twenty-seven specimens of each material were fabricated, randomly divided into three groups of nine and treated as follows--Group 1: stored in distilled water at 37 degrees C for three weeks (control); Group 2: treated with carbamide peroxide (Opalescence Quick) for 30 minutes/week for three weeks; Group 3: treated with 35% hydrogen peroxide power bleach (Opalescence Xtra) for 30 minutes/week for three weeks. For Groups 2 and 3, specimens were stored in distilled water at 37 degrees C during the hiatus periods. The treated specimens were subsequently subjected to microhardness testing (load = 500gf; dwell time = 15 seconds). Results were analyzed using ANOVA/Scheffe's test (p<0.05). For all treatment groups, Spectrum was significantly harder than the other materials and Reactmer was significantly harder than Dyract and Fuji II LC. The effects of in-office tooth whiteners on microhardness were material-dependent. No significant difference in hardness was observed between treatment groups for Dyract and Reactmer. For Spectrum and Fuji II LC, specimens treated with Opalescence Quick were significantly harder than those treated with Opalescence Xtra. No significant difference in hardness was observed between the control and bleached groups for all materials. The hardness of resin-modified glass-ionomer cements, hybrid, polyacid-modified and PRG composites is therefore not significantly affected by the use of 35% carbamide peroxide and 35% hydrogen peroxide in-office tooth whiteners. PMID- 11933904 TI - Influence of ZOE temporary restorations on microleakage in composite restorations. AB - This study investigated the influence of zinc-oxide eugenol (ZOE) temporary restorations on microleakage in composite restorations. Class V cavities were prepared on the buccal surfaces of 32 freshly extracted, non-carious human premolars. The teeth were randomly divided into four groups of eight teeth. Specimens in Group 1 (control) received no temporary restoration. Group 2 and 3 specimens were covered with IRM (Type III ZOE cement) mixed at powder:liquid (P:L) ratio of 10g:1g and 10g:2g, respectively. Specimens in Group 4 were covered with poly-carboxylate cement (eugenol-free) mixed at a P:L ratio of 2.85g:1g. The temporary restorations were removed mechanically with an ultrasonic scaler after one week storage in distilled water at 37 degrees C. The preparations were washed and restored with Scotchbond Multi-Purpose Plus and Z100 according to manufacturers' instructions. The restorations were finished, thermally stressed for 400 cycles and subjected to dye penetration testing. Results were analyzed using Kruskal-Wallis and Mann-Whitney tests at a significance level of 0.05. At both enamel and dentin margins, the microleakage associated with Group 3 was significantly greater than for Groups 1, 2 and 4. For Groups 1 and 4, leakage at the dentin margins was significantly greater than in enamel margins. For the groups pretreated with IRM, no significant difference in dye penetration scores was observed between enamel and dentin. Pre-treatment with IRM mixed at a P:L ratio of 10g:2g significantly increased microleakage and is not recommended clinically. PMID- 11933905 TI - Effects of cyclic temperature changes on water sorption and solubility of composite restoratives. AB - This study investigated the effects of cyclic temperature changes on the water sorption and solubility of four commercial composite resins (Silux Plus, Z100, Ariston pHc and Surefil). The methodology was based upon ISO 4049 procedures with modifications for specimen dimension and thermal-cycling. Eighteen disc specimens (10 +/- 1 mm diameter and 1 +/- 0.1 mm thick) were made for each composite and randomly divided into three groups. The specimens were stored in a desiccator maintained at 35 +/- 1 degrees C until a constant mass was achieved and treated as follows: Group 1--stored in distilled water at 356 degrees C for 178 hrs; Group 2--stored in distilled water at 35 degrees C for 173 hours and subjected to five hours of thermal-cycling with an upper temperature of 45 degrees C; and Group 3--stored in distilled water at 35 degrees C for 173 hours and subjected to five hours of thermal-cycling with an upper temperature of 60 degrees C. Mass after treatment was measured and specimens were re-conditioned to constant mass. The volume of the specimens was obtained and water sorption/solubility calculated. Data was analyzed using factorial ANOVA/Scheffe's post-hoc test at significance level 0.05. The effects of thermal-cycling on water sorption was material dependent. Thermal-cycling at an upper temperature of 60 degrees C significantly increased water sorption of Silux Plus. A significant increase in water sorption was also observed when Z100 was thermal-cycled at an upper temperature of 45 degrees C. The water sorption of Ariston pHc and Surefil was not affected by thermal-cycling. Thermal-cycling did not affect the solubility of all composites. For all treatment groups, Surefil had significantly lower water sorption than the other composites evaluated. The water sorption of Z100 and Surefil was significantly lower than Silux Plus and Ariston pHc. PMID- 11933907 TI - Surface finish of a new hybrid aesthetic restorative material. AB - This study compared the surface finish of a new hybrid aesthetic restorative material (Reactmer) over time to four different types of existing materials. The latter included a composite (Spectrum TPH), a compomer (Dyract AP) and conventional (Fuji II) and resin-modified glass ionomer cements (Fuji II LC). Six specimens of each material were fabricated and stored in distilled water at 37 degrees C for one week. The materials were subsequently finished with a series of Sof-Lex contouring and polishing disks. The average surface roughness (Ra, microm) of each specimen was measured at three days and three months by a surface profilometer. Storage medium was distilled water at 37 degrees C during the hiatus periods. Data was analyzed by ANOVA/Scheffe's and independent samples t tests at significance level 0.05. At both time periods, Fuji II and Fuji II LC were significantly rougher than Spectrum, Dyract and Reactmer. For all materials, surface roughness at three days was not significantly different from that at three months. The surface finish of the giomer (Reactmer) was significantly better than conventional/resin-modified glass ionomer cements and comparable to the composite and compomer evaluated. The quality of surface finish for all materials was not significantly affected by long-term storage in water. PMID- 11933906 TI - The influence of a dentin desensitizer on the microtensile bond strength of two bonding systems. AB - A laboratory study evaluated the influence of a dentin desensitizer (D/Sense 2) on the microtensile bond strength of two adhesive systems: a self-etching primer (Bistite II SC) and a one-bottle adhesive (Prime & Bond 2.1). Sixteen crown dentin discs were obtained from extracted sound human third molars. Dentin surfaces were ground with 600 grit silicon carbide (SiC) abrasive papers to produce a standardized smear layer. Teeth were randomly divided into four groups (n=4). G1-D/Sense 2 + Prime & Bond 2.1; G2-D/Sense 2 + Bistite II SC; G3- and G4 dentin surfaces were bonded with Prime & Bond 2.1 and Bistite II SC, respectively, with no previous treatment with D/Sense 2. Eight mm high resin composite crowns (TPH Spectrum) were incrementally built-up on the treated surfaces. One sample from each group was prepared for evaluation of the hybrid layer on SEM. The specimens for the microtensile test were serially sectioned perpendicular to the adhesive layer to obtain 1 mm2 bounded sticks. Then, each stick was submitted to the microtensile test performed at a 0.5 mm/min crosshead speed. One-way ANOVA and Tukey test showed statistically significant differences among the groups (p<0.05). Values in MPa were: G1-17.85; G2-9.88; G3-35.16; G4 15.57. Based on the results of this study, it can be concluded that the D/Sense 2 desensitizer decreased the bond strength of Prime & Bond 2.1 and Bistite II SC bonding systems. PMID- 11933909 TI - Effect of feeding Cuminum cyminum fruits, Thymus vulgaris leaves or their mixture to rats. AB - Cuminum cyminum fruits and T. vulgaris leaves were fed to male Wistar rats at 2% or 10% of standard diet for 6 w. A mixture (5% +5%) of the 2 plants was also fed to rats for a similar period. Diets containing 2% C. cyminum fruits, 2% or 10% T. vulgaris leaves were not toxic to rats. Impairment of growth and enterohepatonephropathy were observed in the rats fed a diet containing 10% C. cyminum fruits. These changes were also seen in the rats fed the mixture of the 2 plants and were accompanied by leukopenia, anemia and increases in serum AST activity and urea and by decreased total protein and albumin levels. PMID- 11933908 TI - Effect of the photo-activation method on polymerization shrinkage of restorative composites. AB - This study measured the gap that resulted from polymerization shrinkage of seven restorative resin composites after curing by three different methods. Contraction behavior, according to the specimen region, was also characterized. The materials used for this study were Alert (Jeneric/Pentron, Wallingford, CT 06492, USA), Surefil (Dentsply Caulk, Milford, DE 19963, USA), P60 (3M Dental Products, St Paul, MN 55144, USA), Z250 (3M), Z100 (3M), Definite (Degussa-Huls, Hanau, Germany) and Flow-it (Jeneric/Pentron). The composite was placed in a circular brass mold 7 mm in diameter and 2 mm in height. Photo-activation was performed by a) continuous light (500 mW/cm2) for 40 seconds; b) stepped light with low intensity (150 mW/cm2) for 10 seconds and high intensity (500 mW/cm2) for 30 seconds and c) intermittent light (450 mW/cm2) for 60 seconds. The top and bottom surfaces were then polished and after 24 +/- 1 hours, the contraction gap was measured by SEM at variable pressure (LEO 435 VP, Cambridge, England). Results were analyzed by ANOVA and the means compared by Tukey's test (5%). The results demonstrated 1) the continuous light method presented the greatest gap values (15.88 microm), while the other methods demonstrated lower polymerization shrinkage values (stepped light, 13.26 microm; intermittent light, 12.79 microm); 2) restorative composites shrunk more at the bottom surface (15.84 microm) than at the top surface (12.11 microm) and (3) the composites Alert (12.02 microm), Surefil (11.86 microm), Z250 (10.81 microm) and P60 (10.17 microm) presented the least contraction gaps, followed by Z100 (15.84 microm) and Definite (14.06 microm) and finally Flow-it (23.09 microm) low viscosity composite, which had the greatest mean value. PMID- 11933910 TI - Marian Weinbaum Fischman, 1939-2001. PMID- 11933911 TI - Psychological adjustment in the children of mothers with a history of eating disorders. AB - In order to test the impact of mothers' eating disorders (EDs) on their children's psychological adjustment, we recruited mothers belonging to three different populations: women with eating disorders, women with depression, and normal controls. The parents responded to self-report inventories relating to psychological adjustment of the parent and child. The study found that the psychological adjustment of the children of mothers with a history of ED was not different from that of the children of mothers in the normal control group, although mothers described significant pregnancy and birth complications, parenting stress, and symptoms of clinical depression. The children of mothers with a history of depression had significantly greater psychological problems in comparison with those of the children of mothers in the other two groups. The results are interpreted in the context of the protective factors that may have buffered the effects of maternal psychopathology in children of mothers with a history of ED. PMID- 11933912 TI - Are the general equations to predict BMR applicable to patients with anorexia nervosa? AB - AIM: To determine whether the general equations to predict basal metabolic rate (BMR) can be reliably applied to female anorectics. INDIVIDUALS AND METHODS: Two hundred and thirty-seven female patients with anorexia nervosa (AN) were divided into an adolescent group [n=43, 13-17 yrs, 39.3+/-5.0 kg, body mass index (BMI) (weight/height) 15.5+/-1.8 kg/m2] and a young-adult group (n=194, 18-40 yrs, 40.5+/-6.1 kg, BMI 15.6+/-1.9 kg/m2). BMR values determined by indirect calorimetry were compared with those predicted according to either the WHO/FAO/UNU or the Harris-Benedict general equations, or using the Schebendach correction formula (proposed for adjusting the Harris-Benedict estimates in anorectics). RESULTS: Measured BMR was 3,658+/-665 kJ/day in the adolescent and 3,907+/-760 kJ/day in the young-adult patients. In the adolescent group, the differences between predicted and measured values were (mean+/-SD) 1,466 529 kJ/day (+44+/-21%) for WHO/FAO/UNU, 1,587+/-552 kJ/day (+47+/-23%) for the Harris Benedict and -20+/-510 kJ/day for the Schebendach (+1+/-13%), while in the young adult group the corresponding values were 696+/-570 kJ/day (+24+/-24%), 1,252+/ 644 kJ/day (+37+/-27%) and -430+/-640 kJ/day (-9+/-16%). The bias was negatively associated with weight and BMI in both groups when using the WHO/FAO/UNU and Harris-Benedict equations, and with age in the young-adult group for the Harris Benedict and Schebendach equations. CONCLUSIONS: The WHO/FAO/UNU and Harris Benedict equations greatly overestimate BMR in AN. Accurate estimation is to some extent dependent on individual characteristics such as age, weight or BMI. The Schebendach correction formula accurately predicts BMR in female adolescents, but not in young adult women with AN. PMID- 11933913 TI - From the Centers for Disease Control and Prevention. Outbreak of bacterial conjunctivitis at a college--New Hampshire, January-March, 2002. PMID- 11933915 TI - JAMA patient page. Basic science research. PMID- 11933914 TI - From the Centers for Disease Control and Prevention. Update: allograft-associated bacterial infections--United States, 2002. PMID- 11933916 TI - Comparative results of coronary intervention in patients with variant angina versus those with non-variant angina. AB - Coronary angioplasty is reported to be feasible and safe in patients with coronary spasm and fixed stenosis. However, the long-term results are not positive. We compared the results of coronary angioplasty in 20 patients with variant angina versus 17 patients with non-variant angina among 231 consecutive patients with vasospastic angina. Coronary angioplasty was performed successfully in all 37 patients without any complications. Stenting for coronary dissection or recoil was performed in 8 patients, directional coronary atherectomy was selected for ostial lesion of left anterior descending coronary artery stenosis in 2 patients, and standard balloon angioplasty was performed in 27 patients. There were no clinical differences between the two groups. The restensois rate in patients with variant angina was similar to that in patients with non-variant angina (30% vs 29%, ns). There was no relationship between the provoked spasm and restenosis. During the follow-up period, no major complications were observed in patients with variant angina or those with non-variant angina. In conclusion, full medication with calcium channel antagonists and isosorbide dinitrate, and treatment by coronary angioplasty including the use of new devices, were useful treatments for patients with coronary vasospasm and significant organic stenosis. There was no difference concerning the results of coronary intervention between the patients with variant angina and those with non-variant angina. PMID- 11933917 TI - Differences in thrombolytic treatment and in-hospital mortality between women and men after acute myocardial infarction. AB - There is evidence for gender differences in the treatment and outcome of acute myocardial infarction (AMI). However, little data exist about these differences in patients from the Arab Middle East. Therefore, we studied the influence of patient gender on the presentation, the use of thrombolytic therapy, and in hospital mortality after AMI in Kuwaiti nationals. This is a retrospective study of all consecutive Kuwaiti patients admitted to the coronary care unit of a university hospital with the diagnosis of AMI between June 1994 and May 1997. A total of 89 women and 267 men were included. Women were older than men and had significantly higher rates of diabetes (72% vs 46%), hypertension (58% vs 33%) and hypercholesterolemia (80% vs 53%). Women were less likely to receive thrombolytic therapy (40% vs 62%, p=0.001). Fewer women were eligible for thrombolytic therapy (50% vs 66%, p<0.05). Of those who were eligible for thrombolysis there was no sex difference in receiving such treatment. The in hospital mortality among women younger than 70 years was 2.5 times higher than among men in the same age group, while there was no difference in mortality between women and men aged 70 years and older. We conclude that women and men with AMI have different clinical characteristics and outcomes following AMI. There was no gender bias for the use of thrombolytic therapy. The higher in hospital mortality in younger women, i.e. less than 70 years, compared to younger men, indicates that younger women with AMI should be considered as a high-risk group. PMID- 11933918 TI - Association of serum antioxidant capacity with coronary artery disease in middle aged men. AB - The possible involvement of oxidative damage in the progression of atherosclerosis has been suggested. There is some evidence that antioxidant therapy may be beneficial for the prevention of coronary heart disease. In this study, we investigated the relationship between coronary artery disease (CAD) and serum antioxidative status by measuring the total antioxidant status (TAS). Other relevant antioxidants, such as retinol, alpha, gamma-tocopherol, ascorbic acid, alpha, beta-carotenoids, erythrocyte glutathione peroxidase (GSH-Px) and oxidative products, were also determined in 31 male CAD patients with angiographically defined CAD and 66 male controls, aged 40-70 years, in a case control study. The TAS levels, ratio and the concentrations of retinol, albumin, total protein and HDL cholesterol were significantly lower in the CAD patients than in the controls (p<0.01), and alpha-tocopherol and alpha/gamma-tocopherol were significantly higher in the CAD patients than in the controls. The TAS level correlated positively with gamma-GTP, GPT, GOT and uric acid (p<0.01). A multiple regression analysis in the CAD patients revealed that the TAS levels correlated most negatively with the number of diseased vessels. The concentrations of carotenoids and GSH-Px, as well as the alpha/gamma-tocopherol ratio were also significantly associated. Although conditional logistic regression analysis suggested low levels of HDL-cholesterol to be a significant coronary risk factor (OR=5.1, 95% CI=1.09-24.3), the TAS level showed no significant independent contribution to CAD. This study demonstrated an association of antioxidant parameters with the atherosclerosis progression, however, it did not confirm antioxidants as an independent risk factor for CAD event. PMID- 11933919 TI - Endomyocardial biopsy findings in 50 patients with idiopathic atrioventricular block: presence of myocarditis. AB - In patients with primary atrioventricular (AV) conduction disturbances, lesions are commonly believed to be limited to the conduction system. In the present study, myocardial lesions were analyzed histologically in patients with AV conduction disturbances to determine the presence of myocarditis. We studied 50 patients with second or third degree AV block in whom the cause was not clear (AVB group). Endomyocardial biopsy was performed from the right ventricle and examined by light microscopy. Tissue specimens also were obtained from 12 normal hearts at autopsy as a control group (N group). The diagnosis of myocarditis was based on the Dallas criteria. The myocyte transverse diameter was 15.4+/-4.1 microm in the AVB group and 11.7+/-3.1 microm in the N group (P<0.01). The fibrosis area ratio also was significantly higher in the AVB group than in the N group (10.1+/-6.7% vs 5.1+/-2.0%). The mean number of lymphocytes per 400-fold magnified field was significantly greater in the AVB group than in the N group (1.9+/-1.6 vs 1.3+/-0.4). In addition, disorganization of the cardiac myocytes was noted in 8 patients in the AVB group (16%), myocyte disarrangement in 39 patients (78%), myocytolysis in 23 patients (46%), and nuclear deformity in 21 patients (42%). Myocarditis was diagnosed in 3 of the 50 patients (6%). The present study demonstrates that myocardial lesions can be detected in a large proportion of patients with AV conduction disturbances. Furthermore, myocarditis is present in 6% of the cases. PMID- 11933920 TI - The effect of propafenone on premature ventricular contractions (PVC): an analysis based on heart rate dependency of PVCs. AB - The effect of 450 mg/day propafenone for two weeks on premature ventricular contractions (PVCs) was studied in combination with an assessment of heart rate (HR) dependency of PVCs using Holter ECG monitoring in patients with more than 720 PVCs per day. The PVC-HR correlation was classified into positive (P), bidirectional (B), and flat and negative (FN) correlation groups. The positive group included only patients in whom PVC frequency increased with a heart rate increase, while the bidirectional group included patients with PVCs whose frequency increased at low heart rates and decreased at high heart rates. The FN group contained both flat (PVC frequency was almost fixed regardless of heart rate changes) and negative (PVC frequency decreased as heart rate increased) correlations. The effectiveness of propafenone was 70% in the positive group and 50% in the nonpositive group which included both bidirectional (67%) and FN (0%) groups, using a > 70% PVC reduction as a criterion of efficacy. From this, we concluded that propafenone is effective in patients showing either positive or bidirectional PVC-HR correlation. The coupling interval (CI) of PVCs was also prolonged by propafenone as a whole. The present study suggests that there are differences in the mechanism of PVC development in patients with flat or negative correlation and those with a positive or bidirectional correlation. Thus, this type of analysis contributes to an understanding of the action of antiarrhythmic agents, and may allow the prediction of their efficacy on PVCs. PMID- 11933921 TI - Clinical usefulness of the atrial double potential at the intercaval region in the right atrium: a new index for inducibility of atrial fibrillation in electrophysiologic studies. AB - The second deflection of the atrial double potential (DP) recorded at the intercaval region is considered to reflect the far-field potential of the left atrium. The conduction via the upper interatrial connection was evaluated utilizing this DP and the relationship between atrial fibrillation (AF) and the conduction via the interatrial connection evaluated. In 30 consecutive patients with the DP at the intercaval region, prolongation in the left atrial activation time during the right atrial extra stimulation was measured at the intercaval region (deltaDP) and the coronary sinus (deltaCS). The difference between deltaDP and deltaCS (deltaDP-deltaCS) was used as an index of inhomogeneity in interatrial conduction. The patients were divided into AF (n=13) and non-AF (n=17) groups in accordance with the inducibility of AF in the electrophysiologic study. The max deltaDP and the max ACS were greater in the AF group than in the non-AF group, i.e., max deltaDP (43+/-19 vs 27+/-17 ms, P=0.021), max deltaCS (35+/-15 vs 21+/-14 ms, P=0.029). The max absolute value(deltaDP-deltaCS) was also greater in t PMID- 11933922 TI - A randomized, double-blind, placebo-controlled, 8-week study to evaluate the efficacy and safety of once daily atorvastatin (10 mg) in patients with elevated LDL-cholesterol. AB - Lowering of serum cholesterol levels by pharmacologic intervention with statins reduces the incidence of cardiovascular events in subjects with and without atherosclerotic manifestations. In an 8-week, randomized, double-blind study we compared the efficacy and safety of the new compound atorvastatin for reducing LDL-cholesterol (LDL-C) with placebo in an Asian patient cohort. Patients with LDL-C between 160 mg/dl and 250 mg/dl were randomly assigned to treatment with 10 mg atorvastatin or placebo once daily for 8 weeks. At the end of weeks 4 and 8 of the randomized phase, the serum concentrations of lipid parameters as well as safety parameters were determined. Fifty-four patients (32 males and 22 females) were enrolled. Twenty-six patients were assigned to the treatment group. The primary end-point, LDL-C, was reduced by 40% and 42% after 4 and 8 weeks of treatment in the atorvastatin treated patients (p<0.001). The reductions in total cholesterol and triglycerides were up to 31% and 23%, respectively. The HDL-C levels increased up to 11% (p=0.043). There were no significant adverse events. Transient increases in CPK levels (10 times) without myalgia were identified in 1 patient. Atorvastatin, 10 mg/day produced significant reductions in LDL-C, total cholesterol and triglycerides and an elevation of HDL-C levels when used as an adjunct to diet in hyperlipidemic patients. The majority of the clinical effects could be attained by week 4. The overall safety profile of atorvastatin was similar to that of placebo. Atorvastatin was considered to be well tolerated in this patient cohort. PMID- 11933923 TI - Depressive mood accompanies hypercholesterolemia in young Japanese adults. AB - Screening of young hypercholesterolemics is important because they are highly susceptible to atherosclerotic diseases. However, in some cases, serum cholesterol level may be elevated temporarily due to stress or other psychological factors. This study examined the effects of mood states on 'persistent' hypercholesterolemia in comparison with 'temporary' hypercholesterolemia among students entering a university. The subjects were 114 untreated first-year students aged 18 to 20 years old. All had been screened positive for hypercholesterolemia (serum total cholesterol > or = 220 mg/dl) upon enrolling in the university. Three months after the screening, they were divided into two groups according to the re-examined serum total cholesterol level; a persistent hypercholesterolemic group (n=41) with >220 mg/dl and a temporary hypercholesterolemic group (n=73) with <220 mg/dl. At that time, they completed the Profile of Mood States (POMS) with tension-anxiety, depression, anger hostility, vigor, fatigue and confusion scales. The POMS depression scores and the female ratio were higher (both p<0.01) and body mass index was lower (p<0.05) in the persistent hypercholesterolemic subjects than in the temporary hypercholesterolemic subjects; the POMS depression scores were still higher (p<0.05) after controlling for the effects of gender, body mass index, and other POMS scales by multiple logistic regression analysis. Depressive mood appeared to relate to hypercholesterolemia when the university students were screened. Assessment of mood states may be important in screening young hypercholesterolemic patients. PMID- 11933924 TI - The repeatability of left ventricular volume assessment by a new ambulatory radionuclide monitoring system during head-up tilt. AB - The precise measurement of changes in left ventricular volume is important to elucidate the mechanisms of neurally mediated syncope. This study was conducted to determine whether or not a brand-new ambulatory radionuclide monitoring system (C-VEST system) can be clinically used to easily and precisely measure left ventricular volume and function in tilt testing. To assess the repeatability of the C-VEST system, 12 healthy volunteers (mean age 24+/-4 years old) underwent 20 minute head-up tilt testing and we measured the temporal changes in left ventricular volume and ejection fraction twice a day (first and second studies). To investigate the changes in the C-VEST measurements and the detector position in the first and second studies, tilt testing was performed with an 80-degree passive tilt, which is the same as the standard procedure used in diagnosing neurally mediated syncope. The coefficient of repeatability for both the C-VEST and detector position was well within the clinical range (coefficient of repeatability in left ventricular volume ranged from 1.7 to 2.8; coefficient of repeatability in the detector position ranged from 2.3 to 3.1). Precise evaluation of the left ventricular volume can be achieved by an ambulatory radionuclide monitoring system in tilt testing. PMID- 11933925 TI - Coronary reperfusion following ischemia: different expression of bcl-2 and bax proteins, and cardiomyocyte apoptosis. AB - The aim of this work was to examine factors that could be involved in the occurrence of apoptosis in rat hearts subjected to coronary occlusion followed by reperfusion. To this end, we studied the expression of the pro- and anti apoptotic factors, bax and bcl-2, respectively, in reperfused ischemic hearts and in hearts injected with bFGF or saline. In anesthetized rats the left coronary artery was occluded for 45 min, the anesthesia withdrawn and the occlusion removed to allow reperfusion; in sham-operated rats the occlusion was omitted. After 4 hours the rats were decapitated and the heart excised. Sections from the left ventricle were stained with anti-bcl-2-antibody and anti-bax-antibody using the TUNEL method which detects apoptosis. Fragmentation of DNA isolated from reperfused ventricles was examined by agarose electrophoresis. In reperfused hearts no bcl-2 staining was observed in the discrete area in which many cardiomyocyte nuclei were stained by the TUNEL method; outside this area staining for bcl-2 was more marked than in sham-operated rats. Sections from reperfused hearts were stained for bax protein over a wide area including the apoptotic region; sham-operated hearts showed little reaction. Staining for bcl-2 was demonstrable in some nuclei in hearts from saline-injected rats; the numbers were unaffected by i. v. bFGF. Ischemia/reperfusion increases the overall expression of both bcl-2 and bax proteins, but bcl-2 is lost from the reperfused area as indicated by TUNEL staining. Accordingly, the ratio of bcl-2 to bax was reduced in the reperfused area, indicating a pro-apoptotic trend. The marked increase in bcl-2 outside the reperfused area could be a mechanism with which to salvage surviving cardiomyocytes. PMID- 11933926 TI - Independent modulation of L-type Ca2+ channel in guinea pig ventricular cells by nitrendipine and isoproterenol. AB - Dihydropyridine (DHP) Ca2+ channel blockers decrease L-type Ca2+ channel current (I(CaL)) by enhancing steady-state inactivation, whereas beta-adrenergic stimulation increases I(CaL) with small changes in the kinetics. We studied the effects of DHP Ca2+ channel blockers on cardiac I(CaL) augmented by beta adrenergic stimulation. We recorded I(CaL) as Ba2+ currents (I(Ba)) from guinea pig ventricular myocytes using the whole-cell patch clamp technique. and compared the effects of nitrendipine (NIT) in the absence and presence of isoproterenol (1 microM, ISO) or forskolin (10 microM, FSK). Maximal I(Ba) elicited from a holding potential of -80 mV were diminished to 69.4+/-13.5% (mean and SE, n=5) of control by NIT (100 nM) and the diminished I(Ba) were increased to 180.3+/-23.2% of control by ISO in the presence of NIT, which was similar to the enhancement seen in the absence of NIT. NIT shifted the V(1/2) of the I(Ba) inactivation curve from -34.6+/-1.9 mV (n=5) to -48.7+/-1.2 mV, enhancing I(Ba) decay with shortening T(1/2) at -10 mV from 164.6+/-24.2 ms (n=7) to 105.4+/-15.2 ms. ISO elicited a small additional shift in the V(1/2) of I(Ba) inactivation in the same direction. ISO and FSK each slowed I(Ba) decay in the absence of NIT, but not in its presence. Thus, beta-adrenergic agonists increase and DHP Ca2+ channel blockers decrease the amplitude of cardiac I(CaL) independently and the kinetics of I(CaL) is determined mainly by the latter when these drugs coexist. PMID- 11933927 TI - Optimization of atrioventricular delay and follow-up in a patient with congestive heart failure and with bi-ventricular pacing. AB - Cardiac function is improved by bi-ventricular pacing in patients with severe reduced cardiac function. Atrioventricular (AV) delay optimization is also important in this therapy. However, the AV delay required to achieve the optimal AV synchrony varied from time to time. We have reported that the critical AV delay that induces diastolic mitral regurgitation (MR) may represent the upper limit of the optimal AV delay. The optimal AV delay can be predicted by a simple method; slightly prolonged AV delay-interval between the end of atrial kick and complete closure of the mitral valve (duration of diastolic MR) at the AV delay setting. [Case] 60 year old Japanese male with dilated cardiomyopathy. He was repeatedly admitted to our hospital due to congestive heart failure. Ejection fraction was 14%. ECG showed complete left bundle branch block and his PQ interval was 0.22 sec. He was dependent on intravenous injections of catecolamine and could not be discharged from the hospital for over one year. Optimal AV delay was predicted as 80 msec during bi-ventricular pacing by our formula. Cardiac output was 4.9, 6.0, 5.1 l/min when the AV delay was set at 50, 80, 110 msec. Cardiac function was improved from NYHA class III to II and he has been relieved from the dependency on intravenous catecholamine injections. AV delay was optimized (70-100 msec) by our method during follow-up for one year. This case indicates that AV delay optimization is important in bi-ventricular pacing. PMID- 11933928 TI - Successful three-dimensional reconstruction using transesophageal echocardiography in a patient with a left atrial myxoma. AB - Two-dimensional echocardiography can provide intracardiac images. However, the cross-sectional images require mental reconstruction to understand a three dimensional intracardiac structure. It is sometimes hard for inexperienced echocardiographers to engage in reconstruction. Thus, three-dimensional echocardiography is potentially beneficial because these images can provide extra information without mental reconstruction. Herein we demonstrate three dimensional reconstruction using transesophageal echocardiography in a patient with a left atrial myxoma. It contributed to clarifying the surgical considerations, including whether the tumor was adhering to the left atrium or the mitral valve. PMID- 11933929 TI - Killing them softly with kindness: euthanasia legislation in the Netherlands. PMID- 11933930 TI - Broadening the debate on cloning and stem cell research. PMID- 11933931 TI - Staying good while playing god--the ethics of breeding farm animals. AB - Modern genetics has given us some very efficient tools with which to alter the characteristics of animals. To date, farm animal breeders have mainly used these tools to increase productivity. Thus, each new generation of farm animals matures faster, yields more milk, or produces more meat or eggs, than the previous one. Despite these apparent benefits, modern farm animal breeding has had severe negative consequences, including effects on the quality of the animals' lives and biodiversity. The aim of this paper is to discuss the goals and consequences of farm animal breeding within an ethical context. First, a description of what has happened to broilers and dairy cattle as breeders have pursued the goal of ever more efficient production is given. Second, the ethical values that ought to underlie future breeding schemes are discussed. It is suggested that there are in fact two very different ethical approaches: the 'quality of life-based' approach and the 'preservationist' approach. A view combining elements from both approaches is advanced. Finally, an example is given of how it is possible, in practice, to pursue an ethically defensible breeding goal without compromising production efficiency. PMID- 11933932 TI - Tower of Babel: variation in ethical approaches, concepts of welfare and attitudes to genetic manipulation. AB - Attitudes to animal biotechnology are diverse, partly because people have different viewpoints and often do not recognize or acknowledge this to be so. First, people adopt different ethical approaches. If an opponent of genetic manipulation says 'I don't like the idea of altering animals' biology' and a proponent replies '...but it is useful', they are failing to communicate, because one is asking whether the action is right or wrong, whereas the other emphasizes the consequences. Another approach focuses on the person carrying out the action. Many people have hybrid views combining elements of these different approaches. Second, people's concepts of welfare vary, emphasizing animal minds, bodies or natures--or a combination of these. A proponent who argues that a particular genetic change will not cause suffering is unlikely to reassure an opponent who puts more emphasis on naturalness than on feelings or health. An improved dialogue, in which people attempt to understand one another's viewpoints, may enable common principles to be established and practical measures to be taken that enable more cooperation in attempts to improve both human and animal welfare. PMID- 11933933 TI - Should you clone your dog? An animal rights perspective on somacloning. AB - This paper use the Missyplicity Project's detailed Code of Bioethics as a starting point for discussion of animal rights perspectives on cloning. Although funded by a couple in order to clone their pet dog, the project has more important collateral goals and forms part of a general line of research, that, if successful, promises enormous clinical benefits to humans. A particular type of animal rights perspective is described and used to evaluate this project. This perspective accepts a 'principle of axiological anthropocentrism' (PAA), according to which only human beings have certain interests, or a kind of value, which is of pre-eminent moral significance. The best-known animal rights views (those of Singer and Regan) are shown to be consistent with the PAA. This perspective also denies that potential characteristics qualify their possessors for the same type of moral respect as actualized characteristics. The balancing of potential benefits against risks to research subjects is discussed and it is concluded that, from the particular ethical perspective adopted in this paper, cloning research of this general type is not particularly problematic; and, given its stringent Code of Bioethics, only an abolitionist perspective could condemn the Missyplicity Project in particular. PMID- 11933934 TI - The welfare problems associated with using transgenic mice to bioassay for bovine spongiform encephalopathy. AB - Prion diseases are fatal neurodegenerative disorders, epitomized by the the recent bovine spongiform encephalopathy (BSE) epidemic in cattle and the emergence of a novel variant of Creutzfeldt-Jacob disease (vCJD) in humans. In prion disease, the agent of infection is believed to be composed of proteinaceous particles, termed prions, which are converted from a normal isoform into a pathogenic isoform during pathogenesis. A bioassay to detect pathogenic prions of BSE in bovine products consumed by humans was unattainable until the development of transgenic mice, due to the significantly lower susceptibility of wild-type mice to BSE. Transgenic mice have now been generated which express the bovine prion protein and are susceptible to BSE. Following an intracerebral injection with brain homogenate of BSE-infected cattle, transgenic mice develop numerous clinical signs of prion disease, including truncal ataxia (inability to coordinate the torso's muscular activity), increased tone of the tail, generalized tremor, and lack of a forelimb extensor response. In this study, the ethical score system devised by Porter (1992) was applied to the BSE bioassay as a tool for identifying welfare issues affecting animals used in the bioassay. We acknowledge that there are limitations to the use of the information arising from the application of the Porter scoring scheme for assessing the justification to proceed with any animal experiment; notwithstanding these problems, however, our application of the Porter model to the BSE bioassay enabled us to identify potential targets for refinement: pain involved, duration of distress and the duration of the experiment. This was despite lenient scoring for the duration of distress and pain experienced by the mice, and optimal scoring for the quality of animal care. The targets identified for refinement are discussed in relation to the method of inoculation, the duration of the bioassay, and the duration of the clinical phase, with the objective of exploring ways of reducing the severity of the bioassay. PMID- 11933935 TI - Bilateral deep brain stimulation (DBS) of the subthalamic nucleus (STN) or the globus pallidus interna (GPi) for treatment of advanced Parkinson's disease. PMID- 11933936 TI - FDG positron emission tomography for evaluating esophageal cancer. PMID- 11933937 TI - Genetic testing for HFE gene mutations related to hereditary hemochromatosis. PMID- 11933938 TI - High-dose lymphoablative therapy (HDLT) with or without autologous stem-cell rescue for treatment of severe autoimmune diseases. PMID- 11933939 TI - Magnetic resonance imaging of the breast: differential diagnosis of a breast lesion to avoid biopsy. PMID- 11933940 TI - Pancreas retransplantation. PMID- 11933941 TI - Transplanting adult patients with hematopoietic stem cells from placental and umbilical cord blood. PMID- 11933942 TI - Wireless capsule endoscopy in obscure digestive tract bleeding. PMID- 11933943 TI - Extracorporeal shock wave treatment (ESWT) for musculoskeletal indications. PMID- 11933944 TI - Research involving human subjects: ethics and law in early 20th century Russia. AB - This paper reports on the ethical and legal basis of human subjects research in Russia in the first half of the twentieth century, a part of medical history that does not appear previously to have been described. It now seems that both Russia and Germany had regulations before World War II, unlike the UK or US. PMID- 11933945 TI - Assisted suicide and human rights. PMID- 11933946 TI - Archdiocese of Philadelphia: protocol for evaluating Catholic health care collaborative relationships. PMID- 11933948 TI - A lethal evolution. PMID- 11933947 TI - L'Chaim and its limits: why not immortality? PMID- 11933949 TI - Predictive testing. PMID- 11933950 TI - New human subjects rule for pregnant women and fetuses. PMID- 11933951 TI - Changes to reporting of serious adverse events. PMID- 11933952 TI - Institution had many steps to follow before resuming research. PMID- 11933953 TI - Universal protection for all human research participants. PMID- 11933954 TI - Agencies said to be slow on research protections reform. PMID- 11933955 TI - Agency says research institution must respond to research subject's allegations. PMID- 11933956 TI - New review burdens will affect many institutional review boards (IRBs). PMID- 11933957 TI - Stem cell research only with consent. PMID- 11933958 TI - Pros and cons of one agency to oversee human research. PMID- 11933959 TI - The National Ethics Commission disbands. PMID- 11933960 TI - The use of Nazi medical experimentation data: memorial or betrayal? PMID- 11933961 TI - The new world of research ethics: a preliminary map. PMID- 11933962 TI - The murky borderland between scientific intuition and fraud. PMID- 11933963 TI - Character vs. codes: models for research ethics. PMID- 11933964 TI - Criminal responsibility, intervening causation and the right to die. PMID- 11933965 TI - A bill of rights for human subjects of research: a proto-draft. PMID- 11933966 TI - Euthanasia and religious belief: the importance of how we frame the questions. PMID- 11933967 TI - Is there a unique Jewish bioethics of human reproduction? PMID- 11933968 TI - Nursing father and nursing mothers: notes toward a distinctive Jewish view of reproductive ethics. PMID- 11933969 TI - Good gene hunting: commercializing safety and efficacy of home genetic test kits. AB - The rapid advancement of genetic identification as a means of diagnosing or identifying the existence of a genetic trait for preventative and reproductive concerns should compel legal experts to determine whether there is sufficient safety and efficacy in genetic tests. Genetic testing has been used by physicians for a number of years to enhance their allopathic practice. Specifically, genetic tests are now being considered as aids in understanding the development and proliferation of certain diseases in society as a whole, and in specific ethnic and familial groups. The purposes of this paper are to: (1) articulate what genetic testing offers, by way of definition and scope; (2) identify statutory and regulatory limitations on the development, distribution, and use of genetic tests; and (3) consider whether the potential availability of home genetic tests should be made accessible to the American public. This paper will further provide a proposed regulation to encourage the development and distribution of home genetic tests. PMID- 11933970 TI - The use of stem cells in biomedical research. Biotechnology Industry Organization (BIO) position statement. AB - On March 22, 1999, Carl B. Feldbaum, President of the Biotechnology Industry Organization (BIO), submitted comments to the National Bioethics Advisory Committee (NBAC) in response to their request for comments on stem cell research. In the statement submitted to Harold Shapiro, PhD (Chair, NBAC), Mr. Feldbaum made clear that BIO members want to ensure that the promise of new therapies and cures from research using stem cells is realized in a responsible and ethical way. In addition, all BIO members are committed to ensuring that every avenue of promising research can be responsibly explored to improve the health of individuals living with currently intractable diseases. PMID- 11933972 TI - The view of religions toward euthanasia and extraordinary treatments in Japan. AB - 388 Japanese religious groups--143 Shinto, 157 Buddhist, 58 Christian and 30 others--were asked to answer questions regarding several forms of euthanasia and extraordinary treatment during the dying process. Passive euthanasia and indirect euthanasia were accepted by around 70% of the respondents. Active euthanasia was favored by less than 20% of them. Christians were less supportive of euthanasia than practitioners of other religions. Shinto and Buddhist corporations advocated "being natural," when medical treatment became futile at the terminal stage. Religionists' views may deepen the discussion of end-of-life issues. PMID- 11933973 TI - Care of PVS patients: Catholic opinion in the United States. PMID- 11933974 TI - Methotrexate, character, and casuistry: a lesson from Machiavelli. PMID- 11933975 TI - The role of morality in health care its dilemmas. PMID- 11933976 TI - The arguments for euthanasia and physician-assisted suicide: ethical reflection. PMID- 11933977 TI - Contraceptive sterilization in Catholic hospitals is intrinsically evil. PMID- 11933978 TI - European Parliament resolution on human cloning. PMID- 11933979 TI - Reflections on the United States National Institutes of Health draft guidelines for research involving human pluripotent stem cells--theological perspective. AB - Since the human embryonic stem cell research involves destruction of human embryos and, therefore, hinges on the fundamental question of the status of the embryo, it is essential to examine this status carefully in order to establish fitting guidelines for research. The US National Institutes of Health has proposed its own guidelines on the matter recently (1999). The document, rooted in current pluralistic perspectives in moral philosophy (or bioethics), is criticised in this paper as morally inadequate. The argumentation of the criticism stems from the theological perspective on human personhood, which focuses on a continuity of personal identity from embryos to adult human beings. An additional concern for the author is the moral complicity in which the research dependent upon the destruction of human embryonic life is sanctioned. PMID- 11933980 TI - How then should we die?: California's "Death with Dignity" Act. AB - The cultural significance of recent changes in medicine and advances in biotechnology can hardly be overstated. Such have stirred fresh interest in the moral foundations of ethical decision-making and lively debate has ensued as well over the basis of human dignity. Largely divorced from the distinctive moral and ethical commitments that once informed and directed medical practice, modern secular notions of bioethics collapse frequently into human philosophical models of rights and justice. The project of Western medicine now continues within the cultural framework of a radical postmodern agenda that calls for the critical deconstruction and absolute relativizing of all knowledge, and the thorough secularization of the public square. Truth, once understood as a fixed expression of a fundamental reality, has been eschewed in favor of personal preference, subjective experience, private interpretation, and radical perspectivism. Perhaps the greatest challenge facing contemporary culture is that of arriving at a clear and convincing consensus on what constitutes moral surety as well as agreement on what knowledge can serve as an adequate foundation for living the moral life. Having abandoned the reality of divine involvement in the creation and sustenance of human life, contemporary culture now toys with what it means to be human without God. Increasingly popular is the view that whether one possesses dignity or not turns on the question of suffering. Under modern parlance it is simply undignified to suffer. Suffering, somehow, is believed to reduce a person to a state that is incompatible with dignity. Therefore, it should come as no surprise that one of the most pressing social issues today involves the effort to legalize physician-assisted suicide--a project based upon the view that people ought to "die with dignity." In California this view is embodied in Assembly Bill 1592, The "Death with Dignity Act." It is intended to establish California as the second state in the United States to legalize physician-assisted suicide. Patterned closely after Oregon's groundbreaking law, the California bill provides for a terminally ill patient to end his or her own life when certain conditions exist. The purpose of this paper is to review the historical development of California Assembly Bill 1592. An evaluation of both the content of this bill and the process by which it was introduced to the California State Assembly will be offered. The student of bioethics will here find that the usual arguments in support of physician-assisted suicide in the United States, as well as euthanasia in the Netherlands, have been employed in support of AB1592; namely suffering and autonomy. Given the rather predictable pattern of argument advanced by advocates of physician-assisted suicide and euthanasia, this discussion will prove useful for those who wish to offer an informed and sensitive Christian response to similar legislative efforts in other states and countries. PMID- 11933981 TI - Cytogenetic analysis of the cSOD microregion in Drosophila melanogaster. AB - This report describes the genetic organization of a euchromatic region on the third chromosome of Drosophila melanogaster extending cytologically from 68A2 to C1, an interval comprising 10 or 11 polytene chromosome bands. The gene for cytoplasmic superoxide dismutase (cSOD) maps within this interval, as does low xanthine dehydrogenase (lxd).--Recessive lethal mutations were generated within the region by ethyl methanesulfonate mutagenesis and by hybrid dysgenesis. These lethals fall into 11 functional groups, which were partially ordered by complementation with deletions having breakpoints within the region. The distribution of dysgenesis-induced mutations in the region is highly nonrandom, the majority being within a single group. The mutability of this gene is comparable to that of singed (sn), a documented "hot-spot" for P-element insertion.--One of the EMS-induced lethals, l-108, fulfills biochemical criteria expected of a hypomorphic allele of cSOD. To our knowledge this is the first such allele recovered of this gene, and it should prove very useful in an analysis of the in vivo function of cytoplasmic SOD. Indeed, it has been demonstrated that cSOD is almost certainly a vital gene. PMID- 11933982 TI - Improved genetic testing: a new impetus toward universal coverage. AB - As the Human Genome Project increases the predictive power of human genetics, emerging gene chip technology and other advances of genetic testing will give more information to people about their genetic predilections. If insurance companies were allowed to use this information, they would set premiums such that many who need life-saving medical treatment would have no access to it. Americans would not accept this disparity; instead, genetic information will likely remain private, making the modern health insurance system unprofitable for companies and thus pushing the United States towards a universal health care system in the near future. PMID- 11933983 TI - Animal farm--the ethics and public policy of xenotransplantation. AB - In 1997, over four thousand people on organ waiting lists lost their lives. The current organ shortage has led to the need to find alternative sources of organs for transplantation. Advances in immunosuppressive therapies and genetics have made xenotransplantation, the transplantation of animal organs into humans, an increasingly viable technology. However, the promise of xenotransplantation is tempered by the unspecified amount of risk that zoonotic pathogens could mutate and infect humans with potentially deadly diseases. PMID- 11933984 TI - Further abuse of sexually abused children. AB - OBJECTIVE: To determine the incidence of re-abuse in children known to have been sexually abused and to find factors that increase the risk of re-abuse. METHOD: The study group consisted of 183 children with substantiated sexual abuse who presented to two children's hospitals' Child Protection Units in Sydney, Australia during 1988 through 1990. At intake, when the children were aged between 5 years and 15 years, data about the child, the family, and the nature of the index sexual abuse were collected. Six years after presentation for the abuse, records of the Department of Community Services were checked to see if any of the young people had been the subject of substantiated notifications for abuse/neglect before and after intake to the study. Predictors of notifications for abuse/neglect after presentation for the index sexual abuse were identified. RESULTS: Of the sexually abused young people, nearly one in three were the subject of subsequent substantiated notifications to the Department of Community Services for some form of child abuse and neglect or behavior which placed them at risk of harm. Later notifications for abuse/neglect were predicted by notifications for emotional abuse before the index sexual abuse (adjusted RR = 4.88, CI: 1.43 to 16.65), severity of the index sexual abuse (p = .03), and the number of changes in the child's primary caregivers before intake (p = .03). Approximately one in six of the sexually abused young people were notified for sexual abuse after intake to the study. One in 10 also had prior notifications for sexual abuse. Sexual abuse notifications after study intake were predicted by caregiver changes before intake (p = .01) and whether or not there were notifications for emotional abuse before the index sexual abuse (adjusted RR = 3.40, CI: 1.05 to 11.02). CONCLUSIONS: Revictimization of children appears to be a marker of ongoing family dysfunction. Intervention in child sexual abuse needs to consider a range of risk factors associated with re-abuse and, in particular, should focus on family functioning if further abuse is to be prevented. PMID- 11933985 TI - Childhood environment of adult psychiatric outpatients in Norway having been bullied in school. AB - OBJECTIVE: The goal of this study was to contrast the childhood environment of adult psychiatric outpatients reporting to have been bullied at school with those who were not. METHOD: One-hundred-sixty consecutive adult outpatients from a psychiatric clinic in Norway completed self-administered questionnaires about their psychosocial environment during childhood and adolescence. The frequency of being bullied was measured with an inventory used in schools. Also, the Parental Bonding Instrument (PBI) and the Childhood Trauma Questionnaire (CTQ) were used. RESULTS: Men who were bullied in childhood tended to grow up without biological fathers. Women who were bullied scored significantly lower on Father Care on the PBI and significantly higher on Emotional Neglect, Emotional and Physical Abuse and Physical Neglect on CTQ than those who weren't. CONCLUSIONS: The findings suggest that to be bullied in school years is associated with characteristic psychosocial features in the environment of early childhood and adolescence. PMID- 11933986 TI - Self-reports of forgetting and remembering childhood sexual abuse in a nationally representative sample of US women. AB - OBJECTIVE: The purpose of this article is to describe patterns of forgetting and remembering childhood sexual abuse (CSA) in a nationally representative sample of US adult women. METHOD: The respondents were a national probability sample of 711 women, aged 26 years to 54 years, residing in noninstitutional settings in the contiguous 48 states. In a 1996 face-to-face interview survey, trained female interviewers asked each respondent whether she had experienced any sexual coercion by family members or nonfamily members while growing up; whether she believed that she had been sexually abused (by family members or others); and whether she had ever forgotten the CSA experiences and, if so, how she had subsequently remembered them. RESULTS: Twenty-one and six-tenths percent of respondents reported having sexually coercive experiences while growing up; of these, 69.0% indicated that they felt they had been sexually abused. More than one-fourth of respondents who felt sexually abused reported that they had forgotten the abuse for some period of time but later remembered it on their own. Only 1.8% of women self-described as sexually abused reported remembering the abuse with the help of a therapist or other professional person. CONCLUSIONS: The findings indicate that, among women who report CSA, forgetting and subsequently remembering abuse experiences is not uncommon. According to the women surveyed, however, very few (1.8%) of those who felt abused recovered memories of CSA with help from therapists or other professionals. As one of the few studies of CSA memories in a nationally representative sample, this study suggests that therapist-assisted recall is not a major source of CSA memories among women in the US general population. PMID- 11933987 TI - Sexual harassment in Jewish and Arab public schools in Israel. AB - OBJECTIVE: Current empirical literature on sexual harassment in schools is mostly based on nonrepresentative samples of middle-class high-school Caucasian female students. Thus the scope of research regarding gender, age, and cultural differences is very limited. This article reports on findings on sexual harassment in Jewish and Arab schools in Israel with regard to gender, age, and cultural differences. METHOD: The study is part of the first national survey on school violence in Israel. The representative sample includes 10,400 students in grades 7 through 11 attending public schools in Israel. Students were asked to report whether they were victims of specific acts of sexual harassment in school during the month before the survey. RESULTS: Overall, 29.1% of the students were victims of at least one act of harassment. The more common acts were to show offensive pictures or to send obscene letters, to take off or to try to take off part of the student's clothing, and to try to kiss a student. The most vulnerable groups are the Arab boys and 8th grade students. Report rates were the lowest among Arab girls. CONCLUSIONS: Sexual harassment is prevalent in Israeli schools. The pattern of victimization is different for boys and girls and for students in Jewish and Arab schools. These patterns are a complex phenomenon that must be considered in the intervention and policy measures addressing sexual harassment at school. PMID- 11933988 TI - Victim, perpetrator, family, and incident characteristics of infant and child homicide in the United States Air Force. AB - OBJECTIVE: The present study describes factors related to fatal abuse in three age groups in the United States Air Force (USAF). METHOD: Records from 32 substantiated cases of fatal child abuse in the USAF were independently reviewed for 60 predefined factors. RESULTS: Males were over-represented in young child victims (between 1 year and 4 years of age) and child victims (between 4 years and 15 years of age) but not in infant victims (between 24 hours and 1 year of age). African-American infant victims and perpetrators were over-represented. Younger victims were more likely to have been previously physically abused by the perpetrator. Perpetrators were predominantly male and the biological fathers of the victims. Infant and young child perpetrators reported childhood abuse histories, while child perpetrators reported the highest frequency of mental health contact. Victims' families reported significant life stressors. Families of young child victims were more likely divorced, separated, or single. Incidents with infants and young children tended to occur without witnesses; incidents with child victims tended to have the victim's sibling(s) and/or mother present. Fatal incidents were more frequent on the weekend, in the home, and initiated by some family disturbance. CONCLUSIONS: Differences among groups in factors related to infant and child homicide across age groups may assist in the development of more tailored abuse prevention efforts and may also guide future investigations. PMID- 11933989 TI - The effect of services on the recurrence of child maltreatment. AB - OBJECTIVE: Building on a previous model of recurrence, this study examined the relationship of services on the hazard of child maltreatment recurrence during CPS intervention for families who were provided continuing intervention following a confirmed index report of physical abuse or neglect. METHOD: This nonconcurrent prospective study selected 434 subject families who met study eligibility requirements from 1181 families randomly selected from the 2902 families who had experienced a substantiated report of child abuse or neglect during the sampling year. Data were collected and coded from archival sources for 5 years following the index report. Each record was coded by two research analysts to increase inter-rater reliability. Data were analyzed with the Cox Proportional Regression Model. RESULTS: Case characteristics that predicted recurrence were: child vulnerability, family stress, partner abuse, and social support deficits. After examining the potential effect of nine service-related variables only attendance at services predicted recurrence while controlling for other variables in the model. Families who were noted to attend the services identified in their service plans were 33% less likely to experience a recurrence of child maltreatment while their case was active with CPS. CONCLUSIONS: Implications of this and other research suggests that actively engaging families in a helping alliance and helping them accept and receive services may reduce the likelihood of future maltreatment. PMID- 11933990 TI - The Parent Threat Inventory: development, reliability, and validity. AB - OBJECTIVE: The goal was to develop a retrospective inventory of parental threatening behavior to facilitate a better understanding of such behavior's role in the etiology of psychological distress. METHOD: Inventory items were developed based on theory and 135 students' responses to a question eliciting examples of threatening parental behavior. Following item development, two additional student samples (n = 200 and n = 603) completed batteries of self-report measures. Responses were used to eliminate unstable or redundant items from the inventory and to examine the inventory's psychometric properties. RESULTS: Factor analysis of the inventory revealed three factors, accounting for 66.2% of variance; this factor structure is compatible with theory, and consistent across maternal behavior scores, paternal behavior scores, and combined maternal and paternal scores. Cronbach's coefficient alphas indicated acceptable internal consistency; Pearson correlation coefficients indicated acceptable 4-week test-retest reliability. Moderate intercorrelations with two retrospective measures of childhood experiences suggested construct validity. Regression analyses demonstrated the ability of the inventory to predict both anxious and depressive symptomatology and lifetime symptoms of anxiety and depressive disorder. Normative data on combined parent scores, maternal scores, and paternal scores are also presented. CONCLUSIONS: Initial psychometric testing of the Parent Threat Inventory (PTI) suggests it is a reliable and valid tool for investigating the developmental antecedents of adult psychological distress. Further research should focus on addressing two limitations: (1) lack of normative and psychometric data on men and women suffering from clinical disorders, and (2) lack of validation by parental reporting. PMID- 11933992 TI - Constrained formation of object representations. AB - Viewers were presented with a rapid sequence of very brief stimulus pairs, each of which consisted of a pictured object followed by a related or unrelated word. The form of relatedness between the picture and word was manipulated across experiments (identical concept, associated concept, ink color of the picture). Recognition memory for the pictures was affected not only by whether or not paired items were conceptually identical or semantically related, but also by whether or not the words named an irrelevant feature, ink color. These results show that sequential items are integrated on the basis of similarity at whatever level is available, so that the stability of the memory representation of one or both items is increased. We propose that a common mechanism may underlie integration, priming, and selective attention. PMID- 11933993 TI - The finger in flight: real-time motor control by visually masked color stimuli. AB - Current theories of dual visual systems suggest that color is processed in a ventral cortical stream that eventually gives rise to visual awareness but is only indirectly involved in visuomotor control mediated by the dorsal stream. If the dorsal stream is indeed less sensitive to color than the ventral stream, color stimuli blocked from awareness by visual masking should also be blocked from guiding fast motor responses. In this study, pointing movements to one of two isoluminant color targets were preceded by consistent or inconsistent color primes. Trajectories were strongly affected by priming, with kinematics implying a continuous flow of color information into executive brain areas while the finger was already moving. Motor effects were more sensitive to color of the primes than were deliberate attempts to identify the primes in forced-choice tasks based on visual awareness. Priming was observed even when masking was complete. PMID- 11933991 TI - Truth and character: sources that older adults can remember. AB - Are age differences in source memory inevitable? The two experiments reported here examined the hypothesis that the type of source information being tested mediates the magnitude of age differences in source memory. In these studies, participants listened to statements made by two different speakers. We compared younger and older adults' source memory in a traditional perceptual source task (memory for voice) and in two affective, conceptually based source tasks (truth of the statements, character of a person in a photo). In both studies, the perceptual and conceptual source information were conveyed in the same manner, as one speaker was associated with one type of information (e.g., female voice speaks truth). Age differences were robust for decisions regarding who said each statement but were negligible or truth or character decisions. These findings are provocative because they suggest that the type of information can influence age related patterns of performance for source-conveyed information. PMID- 11933994 TI - Can the spotlight of attention be shaped like a doughnut? Evidence from steady state visual evoked potentials. AB - Visual attention enables observers to extract and process high-priority information in the visualfield. Controversy remains as to whether or not observers can ignore information that falls within the spatial beam of attention. We used an objective physiological measure, the steady-state visual evoked potential (SSVEP), to investigate this question. A stream of flickering small, uppercase letters was embedded in the center of a stream of large, uppercase letters. A unitary beam would result in no difference of the SSVEP amplitude elicited by the small letter stream when it was attended versus ignored (i.e., when subjects attended the large letter stream). Contrary to this prediction, SSVEP amplitude increased by almost 100% when the small letter stream was attended compared with when it was ignored. The results support the notion that the attentional spotlight can be formed like a doughnut, processing central information differentially depending on whether it is attended or ignored. PMID- 11933995 TI - Inhibitory tagging on randomly moving objects. AB - Inhibitory tagging is a process that prevents focal attention from revisiting previously checked items in inefficient searches, facilitating search performance. Recent studies suggested that inhibitory tagging is object rather than location based, but it was unclear whether inhibitory tagging operates on moving objects. The present study investigated the tagging effect on moving objects. Participants were asked to search for a moving target among randomly and independently moving distractors. After either efficient or inefficient search, participants performed a probe detection task that measured the inhibitory effect on search items. The inhibitory effect on distractors was observed only after inefficient searches. The present results support the concept of object-based inhibitory tagging. PMID- 11933996 TI - Perspective in statements of quantity, with implications for consumer psychology. AB - We demonstrate that presentation of information about quantities, whether expressed in natural language or by using numbers, induces a perspective that influences subsequent processing. Experiment 1 shows this to be true for natural language quantifiers, with negative and positive expressions inducing different perspectives. In Experiment 2, we examined the application of this idea to the specific case of perspectives induced by describing products as containing x% fat or as being x% fat free. We found that the percentage-fat description appears to induce a perspective that is sensitive to the level offat being depicted, with products being judged as less healthy at higher amounts of fat. However, this effect was lessened (Experiment 2) or eliminated (Experiment 3) with the percentage-fat-free description. The experiments suggest the fat-free perspective blocks access to assumptions about healthy fat levels. PMID- 11933997 TI - Ecstasy and agony: activation of the human amygdala in positive and negative emotion. AB - Considerable evidence indicates that the amygdala plays a critical role in negative, aversive human emotions. Although researchers have speculated that the amygdala plays a role in positive emotion, little relevant evidence exists. We examined the neural correlates of positive and negative emotion using positron emission tomography (PET), focusing on the amygdala. Participants viewed positive and negative photographs, as well as interesting and uninteresting neutral photographs, during PET scanning. The left amygdala and ventromedial prefrontal cortex were activated during positive emotion, and bilateral amygdala activation occurred during negative emotion. High-interest, unusual photographs also elicited left-amygdala activation, a finding consistent with suggestions that the amygdala is involved in vigilance reactions to associatively ambiguous stimuli. The current results constitute the first neuroimaging evidence for a role of the amygdala in positive emotional reactions elicited by visual stimuli. Although the amygdala appears to play a more extensive role in negative emotion, it is involved in positive emotion as well. PMID- 11933998 TI - Lightness constancy: a direct test of the illumination-estimation hypothesis. AB - Many models of color constancy assume that the visual system estimates the scene illuminant and uses this estimate to determine an object's color appearance. A version of this illumination-estimation hypothesis, in which the illuminant estimate is associated with the explicitly perceived illuminant, was tested. Observers made appearance matches between two experimental chambers. Observers adjusted the illumination in one chamber to match that in the other and then adjusted a test patch in one chamber to match the surface lightness of a patch in the other. The illumination-estimation hypothesis, as formulated here, predicted that after both matches the luminances of the light reflected from the test patches would be identical. The data contradict this prediction. A second experiment showed that manipulating the immediate surround of a test patch can affect perceived lightness without affecting perceived illumination. This finding also falsifies the illumination-estimation hypothesis. PMID- 11933999 TI - The representations underlying infants' choice of more: object files versus analog magnitudes. AB - A new choice task was used to explore infants' spontaneous representations of more and less. Ten- and 12-month-old infants saw crackers placed sequentially into two containers, then were allowed to crawl and obtain the crackers from the container they chose. Infants chose the larger quantity with comparisons of 1 versus 2 and 2 versus 3, but failed with comparisons of 3 versus 4, 2 versus 4, and 3 versus 6. Success with visible arrays ruled out a motivational explanation for failure in the occluded 3-versus-6 condition. Control tasks ruled out the possibility that presentation duration guided choice, and showed that presentation complexity was not responsible for the failure with larger numbers. When crackers were different sizes, total surface area or volume determined choice. The infants 'pattern of success and failure supports the hypothesis that they relied on object-file representations, comparing mental models via total volume or surface area rather than via one-to-one correspondence between objectfiles. PMID- 11934000 TI - Evolved sex differences in the number of partners desired? The long and the short of it. AB - Do men seek more short-term mates than women? Buss and Schmitt (1993) showed a pattern of mean difference in the ideal number of sexual partners men and women desired over various time frames. We replicated these mean sex differences (e.g., ideal number over the next 30 years: Ms = 7.69 and 2.78 for men and women, respectively), but in both data sets the sampling distributions were highly skewed. In Study 1, we found few sex differences in medians across time frames (e.g., ideal number over the next 30 years: Mdn = 1 for both men and women). In Study 2, most college men (98.9%) and women (99.2%) said they wanted to settle down with one mutually exclusive sexual partner at some point in their life, ideally within the next 5 years. Neither medians in number of partners desired overall before settling down (replicating Study 1) nor medians in short-term partners desired before settling down (Mdn = 0) differed significantly by gender. Rather, men and women concurred: Short-term mating is not what humans typically seek. PMID- 11934001 TI - Intact perception of biological motion in the face of profound spatial deficits: Williams syndrome. AB - Williams syndrome (WS) is a rare genetic disorder that results in profound spatial cognitive deficits. We examined whether individuals with WS have intact perception of biological motion, which requires global spatial integration of local motion signals into a unitary percept of a human form. Children with WS, normal mental-age-matched children, and normal adults viewed point-light-walker (PLW) displays portraying a human figure walking to the left or right. Children with WS were as good as or better than control children in their ability to judge the walker's direction, even when it was masked with dynamic noise that mimicked the local motion of the PLW lights. These results show that mechanisms underlying the perception of at least some kinds of biological motion are unimpaired in children with WS. They provide the first evidence of selective sparing of a specialized spatial system in individuals with a known genetic impairment. PMID- 11934002 TI - Language comprehenders mentally represent the shapes of objects. AB - We examined the prediction that people activate perceptual symbols during language comprehension. Subjects read sentences describing an animal or object in a certain location. The shape of the object or animal changed as a function of its location (e.g., eagle in the sky, eagle in a nest). However, this change was only implied by the sentences. After reading a sentence, subjects were presented with a line drawing of the object in question. They judged whether the object had been mentioned in the sentence (Experiment 1) or simply named the object (Experiment 2). In both cases, responses were faster when the pictured object's shape matched the shape implied by the sentence than when there was a mismatch. These results support the hypothesis that perceptual symbols are routinely activated in language comprehension. PMID- 11934003 TI - Positive emotions trigger upward spirals toward emotional well-being. AB - The broaden-and-build theory of positive emotions predicts that positive emotions broaden the scopes of attention and cognition, and, by consequence, initiate upward spirals toward increasing emotional well-being. The present study assessed this prediction by testing whether positive affect and broad-minded coping reciprocally and prospectively predict one another. One hundred thirty-eight college students completed self-report measures of affect and coping at two assessment periods 5 weeks apart. As hypothesized, regression analyses showed that initial positive affect, but not negative affect, predicted improved broad minded coping, and initial broad-minded coping predicted increased positive affect, but not reductions in negative affect. Further mediational analyses showed that positive affect and broad-minded coping serially enhanced one another. These findings provide prospective evidence to support the prediction that positive emotions initiate upward spirals toward enhanced emotional wellbeing. Implications for clinical practice and health promotion are discussed. PMID- 11934004 TI - Temporal dynamics of reflexive attention shifts: a dual-stream rapid serial visual presentation exploration. AB - We combined a prototypical exogenous cuing procedure with rapid serial visual presentation (RSVP) to provide a precise characterization of the temporal dynamics of reflexive attention shifts. The novel paradigm thus created has several useful properties, most notably that the physical presentation of the target is neither an onset nor a unique event and that temporal precision is provided without the requirement for a speeded response. A biphasic pattern was observed, with early benefits followed by later costs (inhibition of return) at the cued location relative to the uncued location. The finding of inhibition of return in this paradigm disproves the assertion that inhibition of return is merely a reluctance to respond in the target's direction. It may be partly that, but encoding mechanisms linked to attention must also be involved. PMID- 11934005 TI - Visual attention to color: parvocellular guidance of attentional resources? AB - Although transient changes in luminance have been well documented to automatically attract attention to their location, experiments looking at abrupt changes in color have failed to find similar attentional capture. These results are consistent with current theories of the role of the magnocellular (M) and parvocellular (P) streams that postulate that the M stream, which is "color blind," plays the dominant role in guiding attention and eye movements. The experiment reported here used stimuli that contained only information defined by color, and masked residual luminance information with dynamic noise, to assess the capacity of purely chromatic cues to automatically guide spatial attention. Such stimuli were as effective as those containing large luminance signals in guiding attention. To the extent that these purely chromatic signals isolated the P stream, these results suggest that this stream is also capable of automatic attentional capture. Hence, color vision not only aids target identification but also is a strong aid for target detection and localization. PMID- 11934006 TI - The roles of body and mind in abstract thought. AB - How are people able to think about things they have never seen or touched? We demonstrate that abstract knowledge can be built analogically from more experience-based knowledge. People's understanding of the abstract domain of time, for example, is so intimately dependent on the more experience-based domain of space that when people make an air journey or wait in a lunch line, they also unwittingly (and dramatically) change their thinking about time. Further, our results suggest that it is not sensorimotor spatial experience per se that influences people's thinking about time, but rather people's representations of and thinking about their spatial experience. PMID- 11934007 TI - Priming visual face-processing mechanisms: electrophysiological evidence. AB - Accumulated evidence from electrophysiology and neuroimaging suggests that face perception involves extrastriate visual mechanisms specialized in processing physiognomic features and building a perceptual representation that is categorically distinct and can be identified by face-recognition units. In the present experiment, we recorded event-related brain potentials in order to explore possible contextual influences on the activity of this perceptual mechanism. Subjects werefirst exposed to pairs of small shapes, which did not elicit any face-specific brain activity. The same stimuli, however, elicitedface specific brain activity after subjects saw them embedded in schematic faces, which probably primed the subjects to interpret the shapes as schematic eyes. No face-specific activity was observed when objects rather than faces were used to form the context. We conclude that the activity of face-specific extrastriate perceptual mechanisms can be modulated by contextual constraints that determine the significance of the visual input. PMID- 11934008 TI - Do I know you? Processing orientation and face recognition. AB - Recognition performance is impaired when people are required to provide a verbal description of a complex stimulus (i.e., verbal-overshadowing effect), such as the face of the perpetrator in a simulated robbery. A shift in the processing operations that support successful face recognition is believed to underlie this effect. Specifically, when participants shift from a global to a local processing orientation, face recognition is impaired. Extending research on this general topic, the present experiment revealed that verbalization is not a necessary precondition for the emergence of impaired recognition performance. Rather, face recognition can be disrupted by a task (i.e., letter identification) that triggers the activation of a local processing orientation. Conversely, the activation of a global processing orientation can enhance the accuracy offace recognition. The theoretical and practical implications of these findings for recent treatments of verbal overshadowing and memory function are considered. PMID- 11934009 TI - Now you see it, now you don't: explicit versus implicit measures of the personal/group discrimination discrepancy. PMID- 11934010 TI - Mechanisms of carcinogenicity of aryl hydrazines, aryl hydrazides, and arenediazonium ions. AB - Aryl hydrazines carcinogenesis has been studied for over 25 years and remains poorly understood, although most aryl hydrazines are toxic, tumorigenic, or carcinogenic. In this article, aryl hydrazine carcinogenesis is reviewed comprehensively. The relevant chemistry and biochemistry of aryl hydrazines are first addressed and provide the framework for understanding how aryl hydrazines are metabolized, the reactive intermediates that are produced, and the biological reactive intermediates and products that are formed. Issues of DNA damage, mutagenicity, and enzyme activation are next addressed followed by a brief review of aryl hydrazine tumorigenicity studies. Because several related substrates are metabolized to the same intermediates as are aryl hydrazines, they are briefly discussed. The review concludes with a short discussion of the possible mechanism of carcinogenesis by aryl hydrazines. PMID- 11934011 TI - In vitro protective effects of Terminalia arjuna bark extracts against the 4 nitroquinoline-N-oxide genotoxicity. AB - We determined the antimutagenic potential of chloroform, acetone, methanol, methanol+HCl, diethyl ether, and ethyl acetate extracts of Terminalia arjuna bark against the model mutagen 4-nitroquinoline-N-oxide (4-NQO) using the Salmonella/microsome, comet, and micronucleus (MN) tests. Salmonella typhimurium TA100 strain and human peripheral white blood cells were coincubated with various concentrations (from 5 to 500 microg) of the six extracts and 4-NQO (from 0.05 to 2 microg). We found that the 4-NQO mutagenicity was inhibited by more than 70% in the Salmonella/microsome test at the highest nontoxic extract dose of ethyl acetate (50 microg/plate), chloroform (100 microg/plate), acetone, (100 microg/plate), and methanol (500 microg/plate). A less marked antimutagenicity activity (inhibition of about 40-45%) was observed for the acidic methanol and diethyl ether extracts. The comet assay showed that acetone extract (100 microg/mL) was more effective in reducing the DNA damage caused by 4-NQO (ca. 90%), whereas the chloroform, ethyl acetate, and diethyl ether extracts were cytotoxic. In the MN test, the decrease in 4-NQO clastogenicity was observed by testing the mutagen especially with chloroform and ethyl acetate extracts (inhibition about 40-45%). The acetone and methanol extracts showed a less marked activity (33% and 37%, respectively). The results of the present study suggest that T. arjuna bark contains some nonpolar as well as polar compounds with antimutagenic activity against 4-NQO. Several explanations can be suggested, but further investigations are necessary to definitely identify the active compounds. PMID- 11934012 TI - Modulatory effect of phenolic fractions of Terminalia arjuna on the mutagenicity in Ames assay. AB - We determined the antimutagenicity of phenolic fractions of Terminalia arjuna (soluble and insoluble in chloroform) against two direct-acting mutagens, 4-nitro o-phenylenediamine (NPD) and sodium azide, and against the S9-dependent mutagen 2 aminofluorene (2AF), in TA98 and TA100 tester strains of Salmonella typhimurium. We found that the phenolic fractions of T. arjuna inhibited revertants induced by the S9-dependent mutagen more remarkably than the direct-acting mutagens. Furthermore, the phenolic fractions showed maximum inhibition of 98% and 101.55%, respectively, in the pre-incubation mode of treatment against the mutations induced by 2AF. Overall, the fractions inhibited the revertants induced by S9 dependent mutagens more effectively than those induced by direct-acting mutagens. The percentage of inhibition was higher in the pre-incubation than with direct acting mutagens. The fraction insoluble in chloroform showed more inhibition than the soluble one, which corresponds to a higher polyphenol content in the insoluble fraction than in the soluble extract. PMID- 11934013 TI - DNA single strand breaks induced by low levels of occupational exposure to styrene: the gap between standards and reality. AB - Styrene is a known mutagen and suspected carcinogen, used in the reinforced plastic industry. This study aims to identify the occurrence of DNA single strand breaks (SSBs) in workers exposed to styrene levels far below the recommended standards. We compared 26 exposed workers with 26 control subjects and found a significant increase in the incidence of DNA-SSBs in the exposed individuals. The levels of the biological indices of exposure (urinary mandelic and phenyl glyoxylic acids) were less than 25% of the recommended limits. Reduction of the threshold limit values/time-weighted-average (TLV-TWA) applied is strongly recommended. PMID- 11934014 TI - Studies on sister chromatid exchanges in peripheral lymphocytes of spray painters. AB - In a comparative study of sister chromatid exchanges (SCEs) in cultured lymphocytes, we evaluated the genotoxic risk in 104 male spray painters employed in repair workshops in Chennai City, India, and 50 matched healthy, unexposed controls. We found a higher frequency of SCEs among painters (3.74 +/- 0.11, mean +/- SE) than among controls (2.15 +/- 0.08), and among smoking painters (4.03 +/- 0.21) than among nonsmoking painters (3.55 +/- 0.13), with no significant difference in controls (smokers: 2.1 +/- 0.2; nonsmokers: 2.2 +/- 0.1). Alcoholism did not contribute to an increased SCE frequency. Stepwise multiple linear regression analysis on painters showed that duration of service, smoking, and alcoholism significantly affected SCE scores and explained the 14% variation observed. PMID- 11934015 TI - Expression of telomerase reverse transcriptase in radiation-induced chronic human skin ulcer. AB - The objective of this study was to investigate the expression of the catalytic subunit of telomerase, telomerase reverse transcriptase (TRT), and the possible relationship between the TRT expression and poor healing or cancer transformation in radiation-induced chronic human skin ulcer. Rabbit antibody to human TRT and SP immunohistochemical method were used to detect TRT expression in 24 cases of formalin-fixed, paraffin-embedded chronic human skin ulcer tissues induced by radiation, 5 cases of normal skin, 2 of burned skin, and 8 of cancer. The positive rate of TRT expression in chronic radiation ulcers was 58.3% (14/24), of which it was strongly positive in 41.7% cases (10/24) and weakly positive in 16.7% (4/24). TRT expression was 0% in normal (0/5) and burned skin (0/2), and 100% in cancer cases (8/8). The strongly positive expression of TRT was observed almost always in the cytoplasm and nucleus of squamous epithelial cells of the epidermis but it was negative or only weakly positive in the smooth muscle and endothelia of small blood vessels and capillaries, and in fibroblasts. Chronic inflammatory cells, plasmacytes, and lymphocytes were weakly positive for TRT. TRT expression could be involved in the poor healing caused by sclerosis of small blood vessels and lack of granulation tissue and in the cancer transformation of chronic radiation ulcer. PMID- 11934016 TI - Expression of MMP1 in surgical and radiation-impaired wound healing and its effects on the healing process. AB - Radiation-impaired wound is characterized by delayed healing, nonhealing, and carcinogenesis. The mechanism remains unclear. Matrix metalloproteinases (MMPs) are one family of key regulators of the process of wound healing. Their abnormal expression plays important roles in the formation of some chronic skin ulcers. The objective of this project was to study the expression of MMP1 in surgical and radiation-impaired wound healing and its effects on the healing process and tissue remodeling. A rat model of radiation-impaired wound healing was used. Routine light microscopy, electron microscopy, immunohistochemistry, and in situ hybridization, all of which enabled the detection of MMP1 expression during the healing process, were performed. The wound healing process was impaired and delayed. In rats receiving 25Gy gamma-ray locally, the irradiated wounds healed 6 days later than the nonirradiated controls. The following changes in MMP1 expression were found: (1) In the early inflammatory phase and in the period of granulation tissue formation, MMP1 expression was only slightly if at all affected in the newly formed epidermis of irradiated wounds compared with controls. Later, the epidermal expression of MMP1 in radiation wounds was comparatively increased following the delay of the healing process. (2) MMP1 expression in irradiated wounds was markedly decreased in fibroblasts, endothelial cells, and macrophages compared with controls. The expression phase was prolonged because of the delay of the healing process. The reduced expression of MMP1 in granulation tissue retards such important processes as cell migration, angiogenesis, and tissue remodeling, thus slowing the healing process. The expression ofMMP1 in the proliferating keratinocytes may help re epithelialization. However, in the late healing period, overexpression of MMP1 in the epidermis may hinder the establishment of basal membrane and the formation of granulation tissue, and affect the tissue remodeling process. PMID- 11934017 TI - Detection of human papillomavirus DNA, serum p53, and p53 antibodies in patients with cervical cancer. AB - Human papillomavirus (HPV) was detected in 85% and 63.6% of patients with invasive cervical cancer and minor cervical abnormalities, respectively. HPV-16 was the dominant type in both groups of women. Because of the high oncogenic potential of HPV-16 and the greater chance of its persistence, a follow-up of cases with minor cervical abnormalities harboring HPV-16 is warranted in order to observe the progression of the lesion. As many as 61.5% of the cases with invasive cervical cancer were found to have higher levels of serum p53 protein than did healthy controls. None of the patients had antibodies against the overexpressed p53. This suggests that, even if mutated, the p53 protein may not be immunogenic in all cases. An inverse relationship between the presence of HPV and the alteration in p53 expression was observed in 71.43% of the cases. This could mean the loss of p53 function as a result of either HPV-E6-mediated degradation or mutation in the p53 gene. PMID- 11934018 TI - The effect of recombinant human tumor necrosis factor-alpha on Ehrlich ascites tumor growth. AB - We studied the antitumor effect of recombinant human tumor necrosis factor-alpha (rhTNF-alpha) on the intraperitoneal (i.p.) growth of Ehrlich ascites tumor (EAT) in Swiss albino male mice. The animals were treated with i.p. injection of rhTNF alpha in doses of 5, 7.5, or 10 microg three times a week for 2 weeks, respectively, starting on the 4th day after the EAT inoculation. The effect of the cytokine was evaluated based on the following parameters: total ascites volume, packed cell volume, total packed cell volume, inhibitory growth rate, cellular population of EAT, morphological EAT cell changes, and mean survival time (MST). RhTNF-alpha in a dose of 5 microg had only a slight effect on MST and inhibitory growth rate (IGR). After a dose of 7.5 microg, an increased IGR (p < 0.01) was observed, but the animals did not live longer than the controls. After 7.5- and 10-microg doses (p < 0.001), the number of cells in EAT decreased significantly and enhanced cellular damage to EAT cells was found. In mice treated with 10 microg, a significant IGR (p < 0.001) was accompanied by enhancement of MST (p < 0.01). Although the 10 microg dose exerted a greater effect compared with the remaining doses, no complete regression was attained. PMID- 11934019 TI - Application of recurrent neural network to predict bacterial growth in dynamic conditions. AB - A combination of a factorial design and two central composite designs was used to assess quantitatively the effects of acid pH (5.6-7.0) or alkaline pH (7.0-9.5) and NaCl (0-8%) variations on the growth of Listeria monocytogenes in a meat broth, at 20 degrees C and lower temperature 10 degrees C. Two principal phenomena were observed when bacteria were submitted to abrupt change of pH and a(w) during growth, whatever the growth temperature: (i) large environmental variations induced a lag phase following the fluctuation, and (ii) the growth continued with a generation time value different from that observed before the change or that associated to the new environment. A dynamic model, based on recurrent neural network (RNN), was developed to describe the growth of L. monocytogenes as a function of temperature and fluctuating conditions of acid pH, alkaline pH and concentration of NaCl. The results showed that the neural network model can be used to represent the complex effects of environmental variable conditions on the microorganism behaviour. PMID- 11934020 TI - Some examples of, and some problems with, the use of nonlinear logistic regression in predictive food microbiology. AB - A new technique, nonlinear logistic regression, is described for modelling binomially distributed data, i.e., presence/absence data where growth is either observed or not observed, for applications in predictive food microbiology. Some examples of the successful use of this technique are presented, where the controlling factors are temperature, water activity, pH and the concentration of lactic acid, a weakly dissociating organic acid. Generally speaking, good-fitting models were obtained, as evidenced using various performance measures and goodness-of-fit statistics. As may be expected with a new statistical technique, some problems were encountered with the implementation of the modelling approach and these are discussed. PMID- 11934021 TI - Proposal of a novel parameter to describe the influence of pH on the lag phase of Listeria monocytogenes. AB - Predictive models for lag phase duration (lambda) have been less reliable than specific growth rate (mu) models due, in part, to the influence of the pre-growth environment on lambda. A discrete modelling approach was taken to more completely define the response of individual cells to new environments. Time to detection (td) data was obtained from serial twofold dilutions of Listeria monocytogenes growing in a Bioscreen at 30 degrees C. Comparison of the inoculum densities required to achieve maximum td at growth pH values from 7.2 to 4.7 revealed that, as the growth pH decreased, fewer cells were capable of making the transition to the exponential phase. The proportion of these cells (termed "adaptable cells") in the original inoculum was used to define a new parameter (r0) which, when combined with the constant mean individual cell physiological state parameter (p0), the variation in p0 (SDp0), the inital inoculum (N0) and the maximum population density (Nmax) was able to simulate a complete growth curve. Power transformations with rescaled explanatory variables provided suitable models for the influence of pH on mu, r0, and SDp0, (r2>0.70). PMID- 11934022 TI - Growth pH does not affect the initial physiological state parameter (pO) of Listeria monocytogenes. AB - It has proven difficult to develop adequate mathematical models for the lag phase (lambda) which characterizes the adaptation period prior to the initiation of exponential growth by microorganisms. This is due, in part, to our incomplete understanding of the nature of the initial physiological state of cells (defined as h0 or p0 depending on the model), and changes taking place during adaptation. The objectives of the present study were to characterize p0 using data from growth of Listeria monocytogenes in an automated turbidimetric instrument (Bioscreen), and to determine the influence of limiting growth pH. A model was developed for individual cells which combined a continuous adaptation phase (defined by p0) with a discrete step marking the transition to a continuous exponential growth phase (the CDC model). Parameters of the new model were: p0; the specific growth rate (mu); the initial cell number (N0); and the maximum cell density (Nmax). Progressive reduction of the growth pH in the Bioscreen to 4.7 decreased the p. It was noted that the regression lines for all trials at all pH values appeared to have a common x-intercept (20.086+/-1.092), and it was deduced that, when the Bioscreen detection limit (15.07 In cfu well(-1)) was subtracted, the resulting value represented the "true" value for the initial physiological state of the cells. PMID- 11934023 TI - Optimal temperature input design for estimation of the square root model parameters: parameter accuracy and model validity restrictions. AB - As part of the model building process, parameter estimation is of great importance in view of accurate prediction making. Confidence limits on the predicted model output are largely determined by the parameter estimation accuracy that is reflected by its parameter estimation covariance matrix. In view of the accurate estimation of the Square Root model parameters, Bernaerts et al. have successfully applied the techniques of optimal experiment design for parameter estimation [Int. J. Food Microbiol. 54 (1-2) (2000) 27]. Simulation based results have proved that dynamic (i.e., time-varying) temperature conditions characterised by a large abrupt temperature increase yield highly informative cell density data enabling precise estimation of the Square Root model parameters. In this study, it is shown by bioreactor experiments with detailed and precise sampling that extreme temperature shifts disturb the exponential growth of Escherichia coli K12. A too large shift results in an intermediate lag phase. Because common growth models lack the ability to model this intermediate lag phase, temperature conditions should be designed such that exponential growth persist even though the temperature may be changing. The current publication presents (i) the design of an optimal temperature input guaranteeing model validity yet yielding accurate Square Root model parameters, and (ii) the experimental implementation of the optimal input in a computer controlled bioreactor. Starting values for the experiment design are generated by a traditional two-step procedure based on static experiments. Opposed to the single step temperature profile, the novel temperature input comprises a sequence of smaller temperature increments. The structural development of the temperature input is extensively explained. High quality data of E. coli K12 under optimally varying temperature conditions realised in a computer-controlled bioreactor yield accurate estimates for the Square Root model parameters. The latter is illustrated by means of the individual confidence intervals and the joint confidence region. PMID- 11934024 TI - Utilizing luminometry for monitoring growth of Listeria monocytogenes in its liquid or gelified monocultures and cocultures with "acid-only" Lactococcus lactis. AB - The light output of a bioluminescent recombinant strain of Listeria monocytogenes increased parallel with its viable cell counts during the exponential phase of the growth both in aerobic monocultures and mixed cultures, but dropped significantly at the commencement of the stationary phase. Suppression of L. monocytogenes by a nisin-less strain of Lactococcus lactis occurred only as an early induction of the stationary state of the target organism. In low-salt cocultures, an inverse linear correlation was found between the logarithmic initial counts of lactic acid bacteria and the extent of growth of Listeria. Decrease of luminometric activity of the bioluminescent test organism indicate sensitively the transformation of the cells into metabolically less active stationary state as a stress-adaptive response to nutrient depletion, "metabolic crowding", or, inimical processes. PMID- 11934025 TI - Combining fuzzy querying of imprecise data and predictive microbiology using category-based reasoning for prediction of the possible microbial spoilage in foods: application to Listeria monocytogenes. AB - Various predictive models of microbial behavior have been created and extensive data collection has been done by numerous private or public laboratories. However, significant differences between predicted and observed values in foods have been observed and need to be stressed, understood and explained as much as possible. In this paper, we present a software tool (currently at the level of a prototype) able: (i) to store in a database all relevant information expressed on one hand as qualitative or quantitative data and on the other hand as precise or imprecise data; (ii) to retrieve the more relevant information from the database using queries where criteria may be expressed as fuzzy values in order to enhance the flexibility of the search: (iii) to compute, in addition to the nearest data, an estimation of searched values using statistical models. The architecture of this software tool is structured as a category-based reasoning system. Example queries about Listeria monocytogenes (L. monocytogenes) illustrate the functionalities of this tool. PMID- 11934026 TI - Using survival analysis to investigate the effect of UV-C and heat treatment on storage rot of strawberry and sweet cherry. AB - Ultraviolet light and heat treatment are proposed as alternative techniques for the use of chemicals to reduce the development of the spoilage fungi Botrytis cinerea and Monilinia fructigena on strawberry and sweet cherry, respectively, during storage. In order to investigate the effect of both physical techniques on microbial inactivation and on fruit quality, inoculated berries were subjected to different temperatures (40-48 degrees C) and UV-C doses (0.05-1.50 J/cm2). For each condition, 20 berries were used. After the treatment, fungal growth, visual damage (holes, stains) and fruit firmness were evaluated during a period of 10 days. The experimental data were analysed statistically using survival analysis techniques. Fungal growth on strawberries was significantly retarded using UV-C doses of 0.05 J/cm2 and higher. The same treatment had no significant effect when applied to cherries. The highest doses (1.00 and 1.50 J/cm2) had a negative effect on the calyx of the strawberry, causing browning and drying of the leaves. No beneficial effect of a low temperature treatment (40-48 degrees C) on the shelf life of strawberries was observed, but fungal development on cherries was retarded at temperatures of 45 and 48 degrees C. These temperatures caused severe damage on strawberries (soft stains, holes, decreased firmness), but had no influence on the quality of sweet cherries. PMID- 11934027 TI - Analysing the lag-growth rate relationship of Yersinia enterocolitica. AB - A generalised z-value concept has been applied to analyse the relationship between the lag and the growth rate of Yersinia enterocolitica at a range of temperature, atmospheric carbon dioxide and oxygen percentages. The product of the specific growth rate and the lag (the "work to be done" during the lag phase) is found to be independent of temperature. However, it does depend on the CO2 and O2 concentrations, though the effect of oxygen was less noticeable than the effect of carbon dioxide. PMID- 11934028 TI - Stochastic modelling of bacterial lag phase. AB - In order to study the lag distribution of the individual cells in a bacterial population, a stochastic birth model is used in this study. An integral formula is applied to transform the assumed lag distribution into a growth function describing the transition between lag and exponential phase of the cell population. By means of this formula, it is pointed out that traditional viable count curves are not suitable to identify the distribution of individual cells' lag time. PMID- 11934029 TI - Modelling the effect of sublethal injury on the distribution of the lag times of individual cells of Lactobacillus plantarum. AB - The effect of heat stress on subsequent duration of the lag time of individual cells of Lactobacillus plantarum was analysed by flow cytometry. The results show clearly that both the mean and the standard deviation of the distribution of the lag time increased after sublethal heat treatment. The distributions of the lag times or the log lag times of untreated and treated cells, respectively, could be described as extreme value distributions. From these distributions, the distribution of the minimum lag times could be calculated and thus the effect of inoculum size on the apparent lag could be deduced. The results show clearly that the apparent lag time is dependent on the size of the inoculum, especially when the inoculum is sublethally injured. PMID- 11934030 TI - Aspergillus flavus dose-response curves to selected natural and synthetic antimicrobials. AB - The effects of selected concentrations of antimicrobials from natural (vanillin, thymol, eugenol, carvacrol or citral) or synthetic (potassium sorbate or sodium benzoate) origin on Aspergillus flavus lag time inoculated in laboratory media formulated at water activity (a(w)) 0.99 and pH 4.5 or 3.5, were evaluated. Time to detect a colony with a diameter > 0.5 mm was determined. Mold response was modeled using the Fermi function. Antimicrobial minimal inhibitory concentration (MIC) was defined as the minimal required inhibiting mold growth for 2 months. Fermi function successfully captured A. flavus dose-response curves to the tested antimicrobials with a highly satisfactory fit. Fermi equation coefficients, Pc and k, were used to compare antimicrobials and assess the effect of pH. Important differences in Pc and k were observed among antimicrobials, being natural antimicrobials less pH dependent than synthetic antimicrobials. A large Pc value represents a small antimicrobial effect on A. flavus lag time; thus, high concentrations are needed to delay growth. A. flavus exhibited higher sensitivity to thymol, eugenol, carvacrol, potassium sorbate (at pH 3.5), and sodium benzoate (at pH 3.5) than to vanillin or citral. MICs varied from 200 ppm of sodium bcnzoate at pH 3.5 to 1800 ppm of citral at both evaluated pHs. PMID- 11934031 TI - Modelling the growth kinetics of Listeria as a function of temperature, pH and organic acid concentration. AB - The combined effects of temperature, pH and organic acids (lactic, acetic and propionic) on the growth kinetics of Listeria innocua ATCC 33090 were studied. First, a multiplicative model was built assuming independent effects of all environmental factors. Thus, the model was expanded by the inclusion of a novel term describing the effects of interactions on the growth/no growth limits. The proposed approach allows an accurate description of the boundary between growth and no growth of Listeria. PMID- 11934032 TI - Analysis and practical implementation of a model for combined growth and metabolite production of lactic acid bacteria. AB - Next to the traditional application of lactic acid bacteria (LAB) as starter cultures for food fermentations, the use of LAB as protective cultures against microbial pathogens and spoilage organisms in other food production processes gains more and more interest. The inhibitory effect of LAB is mainly accomplished through formation of antimicrobial metabolites. In this paper, the model of Nicolai et al. [Food Microbiol. 10 (1993) 229.], describing cell growth and production of lactic acid, which is the major end-product of LAB metabolism, is investigated. In contrast to classical predictive models, the transition of the exponential growth phase to the stationary phase is obtained through the increasing concentrations of undissociated lactic acid [LaH] and decreasing pH in the environment. To describe the variation in time of [LaH] and pH, a novel, robust calculation method is introduced. The model of Nicolai et al. in combination with the novel method of [LaH] and pH computation is then further applied to an experimental data set of Lactococcus lactis SL05 grown in a rich medium. An accurate description of the measured values of cell concentration, total lactic acid concentration and pH is obtained. PMID- 11934033 TI - A novel area of predictive modelling: describing the functionality of beneficial microorganisms in foods. AB - Predictive microbiology generally focuses on the potential outgrowth of spoilage bacteria and foodborne pathogens in foods. Little attention has been paid to the biokinetics of beneficial foodgrade microorganisms, such as lactic acid bacteria. The latter is commonly used in the food fermentation industry, mainly for the in situ production of the antimicrobial lactic acid to extend the shelf life of the food. Furthermore, many strains show additional industrial potential as novel starter cultures since they produce functional metabolites, such as bacteriocins and exopolysaccharides. The production of these functional metabolites has been demonstrated during in vitro experiments, but in many cases these novel starter cultures seem to be less efficient when applied in a food system. A modelling approach may contribute to a better understanding of the tight relation between the food environment and bacterial functionality. Primary modelling can be applied to fit the experimental data concerning cell growth, sugar metabolism, and the production of functional metabolites for a given set of environmental conditions. This led to conclusions concerning the growth-associated production of bacteriocin and exopolysaccharides, the inactivation of these molecules when cell growth levels off, and a minimum cell concentration to trigger on bacteriocin production. Examples deal with the production of the bacteriocin sakacin K by the natural fermented sausage isolate Lactobacillus sakei CTC 494, and the production of heteropolysaccharides by the yoghurt starter culture Streptococcus thermophilus LY03. Secondary modelling of biokinetic parameters quantifies the production of bacteriocin and exopolysaccharides in function of environmental factors. As an example, the specific bacteriocin production by Lb. sakei CTC 494 decreases with increasing sodium chloride concentrations. Furthermore, since the assessment of functionality is frequently hampered by the nature of the food system, mathematical modelling techniques may help to predict the functional behaviour of novel lactic acid bacteria starter cultures in a food matrix, and hence quantify in situ production. For example, a model may simulate cell growth and exopolysaccharide production of S. thermophilus LY03 in a milk environment, where direct measurements are difficult to perform. PMID- 11934034 TI - Modelling the competitive growth of Listeria monocytogenes and Listeria innocua in enrichment broths. AB - The overgrowth of Listeria innocua in enrichment broths designed for the isolation of Listeria monocytogenes is believed to result from two factors: a selective growth advantage of L. innocua, and/or an inhibitory interspecies interaction. The generation times of 13 isolates of L. innocua and L. monocytogenes were determined in Brain Heart Infusion (BHI) and a variety of enrichment media. No significant differences were found in growth characteristics between either species in the various media, suggesting that the growth advantage of L. innocua in enrichment media was not as significant as previously described. Kinetic analysis of mixed cultures of L. monocytogenes and isolates of L. innocua producing a variety of inhibitory activities demonstrated the possibility of an inhibitory interaction between these two species resulting in the overgrowth of the enrichment culture with L. innocua. Modelling the evolution of the ratio between two populations in an enrichment process was used to analyze the impact of a selective growth advantage in L. innocua in an enrichment process for growth of L. monocytogenes. These findings support the widely held view that an overgrowth of L. innocua in the enrichment process can result from both a selective growth advantage as well as the production of inhibitory compounds. From a practical perspective, these interactions can result in an increase in false negatives. PMID- 11934035 TI - Modelling microbial growth in structured foods: towards a unified approach. AB - Historically, the ability of foods to support the growth of spoilage organisms and food-borne pathogens has been assessed by inoculating a food with an organism of interest, and following its growth over a period of time. Information gained from such challenge tests, together with knowledge of the organoleptic stability of the product, can then be used to determine an appropriate shelf-life for the food. Whilst this approach may be seen as the "gold-standard" of microbiological assessment of food, it is both time-consuming and costly. A major advance to complement challenge testing was the development of predictive modelling, when it was demonstrated that the growth of a wide range of organisms of interest could be quite accurately modelled as a function of only a few environmental parameters primarily temperature, pH and water activity (a(w)), with perhaps other factors such as nitrite, organic acids and oxygen. This approach to predictive microbiology is embodied in software tools such as the UK Food MicroModel and the Pathogen Modeling Program from the USA. Whilst modelling of this form yields accurate predictions of the growth of organisms in the majority of foods, there are occasions when there are discrepancies between the model and the observed growth. These discrepancies are most often described as "fail-safe", i.e. the observed growth is slower than predicted by the model. This paper examines the role of food structure in the development of microbial populations and communities, and describes the methodologies we propose to begin to tackle some of these complex and interlinked issues. PMID- 11934036 TI - Modeling the lag phase of Listeria monocytogenes. AB - An estimate of the lag phase duration is an important component for predicting the growth of a bacterium and for creating process models and risk assessments. Most current research and data for predictive modeling programs initiated growth studies with cells grown to the stationary phase in a favorable pH, nutrient and temperature environment. In this work, Listeria monocytogenes Scott A cells were grown in brain heart infusion (BHI) broth at different temperatures from 4 to 37 degrees C to the exponential growth or stationary phases. Additional cells were suspended in a dilute broth, desiccated or frozen. These cells were then transferred to BHI broth at various temperatures from 4 to 37 degrees C and the lag phase durations were determined by enumerating cells at appropriate time intervals. Long lag phases were observed for cells initially grown at high temperatures and transferred to low temperatures. In general, exponential growth cells had the shortest lag phases, stationary phase and starved cells had longer, frozen cells had slightly longer and desiccated cells had the longest lag phases. These data were from immediate temperature transitions. When a computer controlled water bath linearly changed the temperature from 37 to 5 degrees C over a 3.0- or 6.0-h period, the cells had short lags and grew continuously with declining growth rates. Transitions of 0.75 or 1.0 h had 20-h lag phases, essentially that of immediate transitions. When the transition was 1.5 h, an intermediate pattern of less than 1 log of growth followed by no additional growth for 20 h occurred. PMID- 11934037 TI - Modelling bacterial growth in quantitative microbiological risk assessment: is it possible? AB - Quantitative microbiological risk assessment (QMRA), predictive modelling and HACCP may be used as tools to increase food safety and can be integrated fruitfully for many purposes. However, when QMRA is applied for public health issues like the evaluation of the status of public health, existing predictive models may not be suited to model bacterial growth. In this context, precise quantification of risks is more important than in the context of food manufacturing alone. In this paper, the modular process risk model (MPRM) is briefly introduced as a QMRA modelling framework. This framework can be used to model the transmission of pathogens through any food pathway, by assigning one of six basic processes (modules) to each of the processing steps. Bacterial growth is one of these basic processes. For QMRA, models of bacterial growth need to be expressed in terms of probability, for example to predict the probability that a critical concentration is reached within a certain amount of time. In contrast, available predictive models are developed and validated to produce point estimates of population sizes and therefore do not fit with this requirement. Recent experience from a European risk assessment project is discussed to illustrate some of the problems that may arise when predictive growth models are used in QMRA. It is suggested that a new type of predictive models needs to be developed that incorporates modelling of variability and uncertainty in growth. PMID- 11934038 TI - Risk assessment of hand washing efficacy using literature and experimental data. AB - This study simulated factors that influence the levels of bacteria on foodservice workers' hands. Relevant data were collected from the scientific literature and from laboratory experiments. Literature information collected included: initial bacterial counts on hands and water faucet spigots, bacterial population changes during hand washing as effected by soap type, sanitizing agent, drying method, and the presence of rings. Experimental data were also collected using Enterobacter aerogenes as a surrogate for transient bacteria. Both literature and experimental data were translated into appropriate discrete or probability distribution functions. The appropriate statistical distribution for each phase of the hand washing process was determined. These distributions were: initial count on hands, beta (2.82, 2.32, 7.5); washing reduction using regular soap, beta (3.01, 1.91, -3.00, 0.60); washing reduction using antimicrobial soap, beta (4.19, 2.99, -4.50, 1.50); washing reduction using chlorhexidine gluconate (CHG), triangular (-4.75, -1.00, 0); reductions from hot air drying, beta (3.52, 1.92, 0.20, 1.00); reduction from paper towel drying, triangular (-2.25, -0.75, 0); reduction due to alcohol sanitizer, gamma (-1.23, 4.42) -5.8; reduction due to alcohol-free sanitizer, gamma (2.22, 5.38) -5.00; and the effect of rings, beta (8.55, 23.35, 0.10, 0.45). Experimental data were fit to normal distributions (expressed as log percentage transfer rate): hand-to-spigot transfer, normal ( 0.80, 1.09); spigot to hand, normal (0.36, 0.90). Soap with an antimicrobial agent (in particular, CHG) was observed to be more effective than regular soap. Hot air drying had the capacity to increase the amount of bacterial contamination on hands, while paper towel drying caused a slight decrease in contamination. There was little difference in the efficacy of alcohol and alcohol-free sanitizers. Ring wearing caused a slight decrease in the efficacy of hand washing. The experimental data validated the simulated combined effect of certain hand washing procedures based on distributions derived from reported studies. The conventional hand washing system caused a small increase in contamination on hands vs. the touch-free system. Sensitivity analysis revealed that the primary factors influencing final bacteria counts on the hand were sanitizer, soap, and drying method. This research represents an initial framework from which sound policy can be promulgated to control bacterial transmission via hand contacts. PMID- 11934039 TI - A probabilistic analysis of Clostridium perfringens growth during food service operations. AB - The purpose of this study was threefold: first, the study was designed to illustrate the use of data and information collected in food safety surveys in a quantitative risk assessment. In this case, the focus was on the food service industry; however, similar data from other parts of the food chain could be similarly incorporated. The second objective was to quantitatively describe and better understand the role that the food service industry plays in the safety of food. The third objective was to illustrate the additional decision-making information that is available when uncertainty and variability are incorporated into the modelling of systems. PMID- 11934040 TI - Predictive modelling for packaging design: equilibrium modified atmosphere packages of fresh-cut vegetables subjected to a simulated distribution chain. AB - The impact of temperature fluctuations in a simulated cold distribution chain, typical of commercial practice, was investigated on both the microbial and sensorial quality of equilibrium modified atmosphere (EMA) packaged minimally processed vegetables. The internal O2 concentration of the designed packages could be predicted for the different steps of the simulated distribution chain by applying an integrated mathematical system. The internal atmosphere in the packages remained in its aerobic range during storage in the chain due to the application of high permeable packaging films for O2 and CO2. Spoilage microorganisms were proliferating fast on minimally processed bell peppers and lettuce. Yeasts showed to be the shelf-life limiting group. Visual properties limited the sensorial shelf-life. Listeria monocytogenes was able to multiply on cucumber slices, survived on minimally processed lettuce and decreased in number on bell peppers due to the combination of low pH and refrigeration. Aeromonas caviae was multiplying on both cucumber slices and mixed lettuce, but was as well inhibited by the low pH of bell peppers. Storage temperature control was found to be of paramount importance for the microbial (spoilage and safety) and sensorial quality evaluation of EMA-packaged minimally processed vegetables. PMID- 11934041 TI - Seafood Spoilage Predictor--development and distribution of a product specific application software. AB - To allow shelf-life prediction of a range of products, the Seafood Spoilage Predictor (SSP) software has been developed to include both kinetic models for growth of specific spoilage microorganisms and empirical relative rates of spoilage models. SSP can read and evaluate temperature profile data of different formats and in this way the software is a flexible device for electronic time temperature integration. Predicted values of microbial growth and of remaining product shelf life can be exported from SSP as graphs and tables in ASCII, HTML and eXtensible Mark-up Language (XML) formats and this allows SSP to be used in combination with other programmes. More than 300 people have downloaded SSP and distribution of this software from the internet has been efficient in stimulating the application of predictive microbiology and of mathematical seafood shelf-life models within industry, research, seafood inspection and teaching. PMID- 11934042 TI - A development environment for predictive modelling in foods. AB - Waikato Environment for Knowledge Analysis (WEKA) is a comprehensive suite of Java class libraries that implement many state-of-the-art machine learning/data mining algorithms. Non-programmers interact with the software via a user interface component called the Knowledge Explorer. Applications constructed from the WEKA class libraries can be run on any computer with a web-browsing capability, allowing users to apply machine learning techniques to their own data regardless of computer platform. This paper describes the user interface component of the WEKA system in reference to previous applications in the predictive modelling of foods. PMID- 11934043 TI - The use of predictive microbiology by the Australian meat industry. AB - In Australia, the key regulatory body, the Meat Standards Committee (MSC) of the Agricultural and Resource Management Council of Australia and New Zealand (ARMCANZ) and the Export Meat Industry Advisory Council (EMIAC) have accepted in principle the usefulness of predictive microbiology for science-based regulation. The predictive microbiology approach is being used in a range of areas including hot-boning, distribution of meat, retailing of meat, fermentation, plant breakdowns and extended chilling regimes. PMID- 11934044 TI - Optimisation of the formulation and of the technological process of egg-based products for the prevention of Salmonella enteritidis survival and growth. AB - The aim of this work was to study the survival of Salmonella enteritidis after a pressure treatment in relation to compositive variables (NaCl content, pH), both in model and real systems, the latter consisting in an egg-based mayonnaise type product. Moreover, the fate of the surviving cells of S. enteritidis has been monitored during storage at 10 degrees C and the growth or death parameters have been calculated and modelled in relation to pH, NaCl concentration of the medium and entity of the pressure treatment applied. The modelling of the effects of the environmental factors on the treatment effectiveness indicated that the salt content and pH displayed a synergistic effect with pressure, whose extent was higher in the mayonnaise based products than in BHI. In fact, while in the model systems the cell recovery and growth during the subsequent incubation at 10 degrees C was allowed in many combinations of the Central Composite Design, in the real systems no recovery or growth of S. enteritidis were observed. This viability loss, which was maximum at pH 4.00 or 2% NaCl, could not be attributed merely to the interactions of such variables, but probably involves the naturally occurring antimicrobial enzymes of the raw material, whose activity can be enhanced by the pressure treatment. PMID- 11934045 TI - Application of shelf life decision system (SLDS) to marine cultured fish quality. AB - Growth of natural microflora of marine cultured, air-packed, sea bass (Dichentrachus labrax) was studied at isothermal conditions in the 0-15 degrees C range and kinetically modelled using the four-parameter Logistic equation. Sensory shelf life was correlated to pseudomonad population and sensory acceptability was correlated to a pseudomonad level, Ns, of 10(7). The variability of their initial population was quantitatively shown and a conductance-based rapid method specific to sea bass pseudomonad enumeration was established as a practical means of N0 determination, required in shelf life predictions. Kinetic models, shelf life correlations and N0 data were incorporated into the shelf life decision system (SLDS) shown to be an effective tool for marine cultured sea bass chill chain management leading to optimization of quality of the fish at consumer's end. PMID- 11934046 TI - Characterisation and modelling of oscillatory behaviour related to reuterin production by Lactobacillus reuteri. AB - During reuterin production by Lactobacillus reuteri in a chemostat, the growth, substrate and metabolite concentrations showed oscillatory behaviour. The sensitivity of L. reuteri towards reuterin was shown to be a possible explanation of the oscillatory behaviour. A deterministic mathematical model consisting of four coupled differential equations describing the concentrations of biomass, glucose, glycerol and reuterin with time was developed. With a set of parameter values determined from batch experiments, the model was able to predict both oscillatory and steady state behaviour in a chemostat by changing the input variables. In a batch system, the model was able to give a satisfactory description of the glucose and glycerol concentrations but not of the biomass and reuterin concentrations. Mathematical modelling of the system was shown to be an effective and systematic approach in exploring a complex biological system. PMID- 11934047 TI - Predictive microbiology: towards the interface and beyond. AB - This review considers the concept and history of predictive microbiology and explores aspects of the modelling process including kinetic and probability modelling approaches. The "journey" traces the route from reproducible responses observed under close to optimal conditions for growth, through recognition and description of the increased variability in responses as conditions become progressively less favourable for growth, to defining combinations of factors at which growth ceases (the growth/no growth interface). Death kinetics patterns are presented which form a basis on which to begin the development of nonthermal death models. This will require incorporation of phenotypic, adaptive responses and may be influenced by factors such as the sequence in which environmental constraints are applied. A recurrent theme is that probability (stochastic) approaches are required to complement or replace kinetic models as the growth/no growth interface is approached and microorganisms adopt a survival rather than growth mode. Attention is also drawn to the interfaces of predictive microbiology with microbial physiology, information technology and food safety initiatives such as HACCP and risk assessment. PMID- 11934048 TI - Debating the biological reality of modelling preservation. AB - Predictive food microbiology is a rapidly developing science and has made great advances. The aim is to debate a number of issues in modelling preservation: (1) inoculum and prehistory effects on lag times and process susceptibility; (2) mechanistic vs. empirical modelling; and (3) concluding remarks (the Species concept, methodology and biovariability). Increasing the awareness in these issues may bridge the gap between the complex reality in food microbial physiology and the application potential of predictive models. The challenge of bringing integrated preservation or risk analysis further and developing ways to truly model and link biological susceptibility distributions from raw ingredients via process survival to outgrowth probabilities in the final product remains. PMID- 11934049 TI - Simulation modelling of bacterial growth in yoghurt. AB - INDISIM, an individual-based simulator, was used to specifically study the influence of the shape and size of Streptococcus salivarius subsp. thermophilus and Lactobacillus delbrueckii subsp. bulgaricus in yoghurt processing. To this effect, two different sets of simulations were carried out. In the first set of simulations, it was assumed that the initial acidity of the medium has the same value as the acidity of the cytoplasm of the microorganisms. Hence, the differences in bacterial growth by the two species are only attributable to differences in their geometry. It was found that, in an optimum culture, the growth in biomass of S. salivarius subsp. thermophilus is larger than that of L. delbrueckii subsp. bulgaricus. An important factor for understanding this difference might be the larger mass-to-surface ratio of the former. In the second set of simulations, a simplified model of yoghurt, the parametrisation differs both in the geometry and the metabolism of the two species. The results of these simulations are in very good qualitative agreement with the experimental data of [Lait 69 (1989) 519]. Finally, by inhibiting the uptake of amino acids by S. salivarius subsp. thermophilus, the large relative importance of lactic acid in yoghurt processing was highlighted. PMID- 11934050 TI - Bibliography of food microbiology. PMID- 11934051 TI - Postoperative mandibular stability after orthognathic surgery in patients with mandibular protrusion and mandibular deviation. AB - Skeletal stability and temporomandibular joint (TMJ) signs and symptoms were analyzed in 23 patients in whom mandibular protrusion and mandibular deviation had been corrected using bilateral sagittal split ramus osteotomy (BSSRO group, n = 10) and unilateral SSRO and intraoral vertical ramus osteotomy (USSRO+IVRO group, n = 13). Miniplate fixation was used in SSRO but no fixation was used in IVRO. The ratio of condylar bony change was 30.4% (7/23) and all condylar bony changes were seen on the deviated side. All preoperative signs and symptoms of TMJ disorders (4/13 patients in the USSRO+IVRO group and 2/10 patients in the BSSRO group) disappeared after surgery. Comparing the USSRO+IVRO group and the BSSRO group, in patients without condylar bony change, the mandible in both groups was stable anteriorly and horizontally after surgery, even though there was a larger horizontal mandibular movement in the USSRO+IVRO group during surgery. Comparing patients with condylar bony change versus no condylar bony change in the USSRO+IVRO group, postoperative horizontal mandibular displacement was significantly larger in the condylar bony change group than in the no condylar bony change group. These results support the idea that USSRO+IVRO can be useful in correcting mandibular deviation as well as improving signs and symptoms of TMJ disorders. However, it also seems important to be aware of the possibility of horizontal mandibular relapse in patients with condylar bony change. PMID- 11934052 TI - A retrospective analysis of 20 surgically corrected bimaxillary protrusion patients. AB - This paper presents a retrospective review and analysis of 20 bimaxillary protrusion patients who visited the authors' hospital between 1986 and 1998 following surgical correction. The lateral cephalometric radiographs of each patient were taken preoperatively (T0), within 1 week after surgery (T1), and at least 1 postoperative year later (T2). Hard and soft tissue analysis was performed on each cephalometric radiograph. The matched pair t test was employed for T0-T1, T1-T2, and T0-T2 periods. The sample consisted of 20 Korean adult patients with bimaxillary protrusion (18 women and 2 men), aged 21 to 33 years. The first premolars were removed in 18 of the 20 cases. The Wunderer method was selected in 18 of the 20 maxillary cases, and the anterior subapical osteotomy was selected in all mandibular cases. Augmentation genioplasty was combined in 3 cases, and reduction glossoplasty was combined in 2 cases. Orthodontic treatment was accompanied in 8 cases. The statistical analysis of all the variables revealed that, except for overbites, there were significant differences between T1-T2 and between T0-T2 periods (P < .01). This suggests that most of the bimaxillary patients want instant esthetic facial results and that their soft tissue profiles were improved significantly. However, the postoperative course should be cautiously observed. PMID- 11934053 TI - Nasolabial esthetics after Le Fort I osteotomy and V-Y closure: a statistical evaluation. AB - The Le Fort I osteotomy results in predictable long-term effects. The soft tissues, however, can be difficult to control because of considerable variation in their adaptation. Several adverse reactions can take place, including thinning and lateral retraction of the lip, accentuation of the nasolabial groove, reduced vermilion exposure, and increase of the nasolabial angle. In this study on 51 selected patients, the V-shaped wound in the vestibule of the maxilla after the Le Fort I osteotomy was closed with a "Y," with the bast of the "Y" in the midline of the upper lip (V-Y closure). Forward multiple regression analysis was calculated for each bony landmark. Regression equations were formulated if P < .05. The equation with the bony point with the highest r2 value was considered the most important independent variable. The selected independent variables were used to form 4 subgroups with identical vectors of movements: impaction, advancement, impaction and advancement, and dorsal impaction. In these 4 subgroups, forward multiple regression analysis was used to select equations with the highest r2 value (P < .05). The selected equations demonstrated that a V-Y plasty as single soft tissue procedure suffices only in advancement cases, and only if the nose does not need additional upward rotation. In the other 3 subgroups, additional procedures are necessary, such as alar cinch suture, reduction of the anterior nasal spine, or grinding of the paranasal area in order to prevent the above-mentioned adverse soft tissue changes. PMID- 11934054 TI - Effect of mandibular setback surgery on the posterior airway size. AB - Treatment of dentofacial deformities with jaw osteotomies has an effect on airway anatomy, and therefore, mandibular setback surgery has the potential to diminish airway size. The purpose of this study was to evaluate the long-term effect of mandibular setback surgery on the airway size. The material consisted of pre- and postoperative (minimum 1 year) lateral radiograms of 22 individuals (18 females and 4 males) with a mean age of 30 years, who had undergone mandibular setback surgery to correct skeletal Class III discrepancies. Hard and soft tissue points were digitized with a Numonics Accugrid digitizer and analyzed with Xmetrix software. A paired t test was used to evaluate the difference between pre- and postoperative measurements. In addition, Pearson's coefficient correlation was calculated to reveal the possible association between the skeletal change in relation to the change in airway size. The mean value for the initial SNA was 81.3 degrees, 85.4 degrees for SNB, 36.9 degrees for S-Na/MP, and for the posterior airway, 10.5 mm and 12.0 mm retropalatinally and retrolingually, respectively. At the postoperative evaluation, SNB was 80.7 degrees, S-Na/MP 41.0 degrees, and the posterior airway retropalatinally 8.3 mm and retrolingually 9.8 mm. Statistical analysis revealed a highly significant correlation between the change in the ANB angle and in the S-Na/MP angle versus the change in the upper airway size, both retropalatinally and retrolingually. Mandibular setback surgery with posterior rotation may gradually result in increased upper airway resistance in cases where neuromuscular adaptation is insufficient to compensate for the reduction in the airway size. Therefore, large anteroposterior discrepancies should be corrected by combined maxillary and mandibular osteotomies. PMID- 11934055 TI - Extrusion-based leveling with segmented arch mechanics. AB - Leveling of the dental arch may be accomplished through anterior intrusion, posterior extrusion, or a combination. Posterior extrusion is usually preferred in vertically balanced adolescents and in surgical-orthodontic treatment of adults suffering from deep bite and mandibular deficiency. A major disadvantage of posterior extrusion, when accomplished by continuous archwires, is incisor flaring. Here a segmented arch arrangement is proposed for posterior extrusion that combines the clinical simplicity of continuous mechanics with better control of incisor position. PMID- 11934056 TI - Three-dimensional hard tissue palatal size and shape: a 10-year longitudinal evaluation in healthy adults. AB - A 10-year longitudinal evaluation of the morphology (size and shape) of hard tissue palate was performed in 6 female and 6 male healthy adults (mean age at the second evaluation was 33 years, SD = 2.2). All subjects had a complete permanent dentition, including the second molars, and were free from respiratory problems. Palatal landmarks were digitized with a computerized 3D instrument, and their coordinates were used to derive a mathematical model of palatal form. Palatal shape (size-independent) was assessed by a fourth-grade polynomial in the sagittal and frontal plane projections. Palatal dimensions in the frontal and sagittal planes were computed and compared between the 2 evaluations by paired Student t tests. A great variability was observed, and no significant modifications in size were found (P > .05 for all variables). No variations in shape were observed. Sex had no significant effect for any variable (Student t for independent samples, P > .05). This study showed that in healthy subjects, hard tissue palatal morphology does not seem to change between the third and the fourth decades of life. PMID- 11934057 TI - Description of dental arch form using the Fourier series. AB - The aim of this study was to describe the form of the human superior dental arch using Fourier transformations. Forty models made in dental stone from impressions of the maxillary dental arch were used to obtain the reference data, which were expressed in Cartesian coordinates, from the mesovestibular cuspid vertices of molar teeth, vestibular cuspid of premolars, and incisal edge. Fourth-grade equations and Fourier series were calculated from these data. The results indicate that Fourier series more precisely express the form and size of different dental arches, with mixed or permanent dentition, than fourth-grade equations. Details of the mathematical procedure and the precision obtained were provided. PMID- 11934058 TI - Readiness for orthognathic surgery: a survey of practitioner opinion. AB - The purpose of this study was to determine if a general consensus exists among orthodontists regarding the features of an adequate presurgical setup. A questionnaire was developed to assess the importance of 12 criteria for surgical readiness. Each criterion was rated on a 5-point scale (1 = Unimportant, 5 = Critical). This questionnaire was sent to 104 randomly selected, board-certified orthodontists in the United States and Puerto Rico. Usable questionnaires were returned by 57 respondents (55% response rate). The highest and most consistent ratings were given for Arch Compatibility in the transverse dimension (mean = 4.66, SD = 0.55), Crossbite (mean = 4.57, SD = 0.63), and Torque of the Anterior Teeth (mean = 4.38, SD = 0.73). Further analysis showed a correlation between surgical experience of the orthodontist and a higher rating of importance for Torque of the Mandibular Teeth (Spearman r = 0.38), Torque of the Maxillary Teeth (Spearman r = 0.37), and Torque of the Anterior Teeth (Spearman r = 0.28). These results suggest that as orthodontists gain surgical experience, they attribute more importance to correcting the torque of the teeth. Also, the relatively large differences in opinion regarding presurgery orthodontic setup support a need for further research and/or educational effort regarding what criteria are important before surgery. PMID- 11934060 TI - Preliminary results on the safety and efficacy of the OssaTron for treatment of plantar fasciitis. AB - The OssaTron may be another alternative for management of plantar fasciitis (heel pain syndrome) after failure of non-operative management and prior to surgical management. This study evaluated primarily the safety and early preliminary efficacy of the OssaTron in treatment of patients with plantar fasciitis unresponsive to non-operative management. Twenty heels of 20 patients were treated with 1000 extracorporeal shockwaves from the OssaTron to the affected heel after administration of a heel block. The patients were followed for one year. Each patient was evaluated by roentgenogram, KinCom, range of motion and physical examination, including evaluation of point tenderness by means of a palpometer and according to a 10-cm visual analog scale. The control was the contralateral heel. Patients also performed self evaluation by means of patient activities of daily living questionnaire and pain reported by a 10-cm visual analog scale. There were no complications or adverse effects attributed to the procedure of orthotripsy. Of the 20 patients treated, 18 were improved or pain free. Eighteen of the 20 subjects treated stated that they would undergo the procedure again instead of surgery. Based on these results, we concluded that orthotripsy is a safe and effective method of treating heel pain syndrome that has been unresponsive to nonoperative management. PMID- 11934061 TI - Shockwave therapy for patients with plantar fasciitis: a one-year follow-up study. AB - The effect of shockwave therapy was investigated in 79 patients (85 heels) with plantar fasciitis with one-year follow-up. There were 59 women and 20 men with an average age of 47 (range, 15-75) years. Each patient was treated with 1000 impulses of shockwave at 14 kV to the affected heel. A 100-point scoring system was used for evaluation including 70 points for pain and 30 points for function. The intensity of pain was based on a visual analogue scale from 0 to 10. The overall results were 75.3% complaint-free, 18.8% significantly better, 5.9% slightly better and none unchanged or worse. The effect of shockwave therapy seemed cumulative and was time-dependent. The recurrence rate was 5%. There were no device-related problems, systemic or local complications. Shockwave therapy is a safe and effective modality in the treatment of patients with plantar fasciitis. PMID- 11934062 TI - Nerve entrapment in painful heel syndrome. AB - Subcalcaneal heel pain is one of the most common foot ailments, yet the exact etiology is still controversial. Nerve entrapment has been suggested as one of the possible causes of this painful condition in recalcitrant cases. The purpose of this study is to determine the role of nerve entrapment in painful heels. Twenty patients with heel pain (25 heels) were compared with an age and body mass index-matched control population using electrodiagnostic methods. The results of the study revealed 22 heels (88%) with heel pain had lateral plantar nerve entrapment signs with or without medial plantar nerve findings on EMG. There were no abnormal values in the control group. Nerve entrapment syndrome has previously been considered only in cases with intractable heel pain, but this study suggests that it may play a role the early phases of painful heel syndrome. PMID- 11934063 TI - Heel pain triad (HPT): the combination of plantar fasciitis, posterior tibial tendon dysfunction and tarsal tunnel syndrome. AB - Between 1996 and 1999, we evaluated 286 patients with chronic heel pain. We identified 14 patients who were diagnosed and surgically treated for a unique combination of plantar fasciitis, posterior tibial tendon dysfunction and tarsal tunnel syndrome. We postulate that failure of the static (plantar fascia) and dynamic (posterior tibial tendon) support of the longitudinal arch of the foot has resulted in traction injury to the posterior tibial nerve, i.e., tarsal tunnel syndrome. The combination of plantar fasciitis, posterior tibial tendon dysfunction and tarsal tunnel syndrome was recognized and treated. We have called this combination the "Heel Pain Triad (HPT)." Using the AOFAS hindfoot rating system, retrospective chart review and patient examination revealed marked improvement in 85.7% of patients. Follow-up was done four to 33 months (mean follow-up was 17.1 months). Marked improvement was noted in the categories of pain, activity level, walking distance, walking surface and limp. Improvement was statistically significant for all categories. PMID- 11934064 TI - The SCARF osteotomy for the correction of hallux valgus deformities. AB - The authors report their experience with a modified SCARF osteotomy with three years follow-up. Correction of moderate to severe hallux valgus deformities was achieved using a Z step osteotomy cut to realign the first metatarsal bone. A retrospective analysis was undertaken in 89 consecutive patients (111 feet). Results were analyzed by clinical examination, a questionnaire including the AOFAS forefoot score, and plain roentgenograms. Hallux valgus and intermetatarsal angle improved at mean 19.1 degrees and 6.6 degrees, respectively. Mean forefoot score improved from 50.1 to 91 points out of 100 possible points. Satisfactory healing time was expressed by an average return back to work of 5.8 weeks and back to sport of 8.3 weeks. Persistence or recurrence of hallux valgus was seen in seven patients (6%). The complication rate was 5.4% including superficial wound infection, traumatic dislocation of the distal fragment, and hallux limitus. The presented technique provides predictable correction of moderate to severe hallux valgus deformities. PMID- 11934065 TI - Metatarsal strains are sufficient to cause fatigue fracture during cyclic overloading. AB - Human in vivo tibial strains during vigorous walking have not been found to exceed 1200 microstrains. These values are below those found in ex vivo studies (>3000 microstrains) to cause cortical bone fatigue failure, suggesting that an intermediate bone remodeling response may be associated with tibial stress fractures. Metatarsal stress fractures, however, often develop before there is time for such a response to occur. Simultaneous in vivo axial strains were measured at the mid diaphysis of the second metatarsal and the tibia in two subjects. Peak axial metatarsal compression strains and strain rates were significantly higher than those of the tibia during treadmill walking and jogging both barefoot and with running shoes and during simple calisthenics. During barefoot treadmill walking metatarsal compression strains were greater than 2500 microstrains. During one- and two-leg vertical jumps and broad jumping, both metatarsal compression and tension strains were >3000 microstrains. Compression and tension strains in the metatarsus unlike those of the tibia may be sufficiently high even during moderate exertional activities to cause fatigue failure of bone secondary to the number of loading cycles without an intermediate bone remodeling response. PMID- 11934066 TI - The shape of the metatarsal head as a cause of hallux abductovalgus. AB - The curvature of the 1st metatarsal head was evaluated on 100 radiographs by calculating the functional angle of the joint surface. This measure of curvature varied positively with the degree of hallux abductovalgus (HAV). Using measures of correlation it was found that in males, the association was linear (r2=0.39, p<0.001). In females, a linear relationship was identified only after logarithmic transformation of the data (r2=0.25, p<0.001). Comparison of the functional angle between males and females showed a significant difference (t=-5.03, df=98, p<0.001). PMID- 11934067 TI - Revision ankle arthrodesis. AB - Between 1996 and 1999, we performed successful revision ankle arthrodesis for aseptic nonunion in 10 patients using external fixation with bone graft and an implantable bone stimulator. The etiology of the ankle arthrosis in the patients was post-traumatic in eight and rheumatologic in two. The average age of the patients was 54. On average, each patient had received 2.5 previous surgeries (range, 1 to 5) prior to the original fusion attempt. Fusion occurred in all 10 of these patients at an average of 12.8 weeks following revision (range, nine to 20 weeks). Complications occurred in two patients: one had a pin tract infection which resolved with local pin care and oral antibiotics; the second patient had a varus malalignment which was accommodated with orthotics. This patient also had a deep peroneal nerve neuropraxia which resolved with observation. We evaluated all 10 patients with clinical and radiographic examination, and determined their functional results for a duration of follow-up of 15 months (range, six to 36 months). Using a modification of the AOFAS ankle/hindfoot scoring system, the results were: three excellent, four good, two fair, and one poor. All but one of these patients were satisfied with the outcome of this procedure. Radiographic analysis supported good clinical alignment and solid fusion. PMID- 11934068 TI - Comparison of two methods used to assess first-ray mobility. AB - Mobility of the first-ray is associated with several common lower extremity disorders. However, the reliability and validity of clinical measurement remains unclear. In this study we examined first-ray mobility by using one hand to stabilize the lesser metatarsals while the clinician's other hand applied a displacement force to the head of the first metatarsal. The amount of mobility was graded as stiff, normal or hypermobile. We then used a well-validated mechanical device to perform similar tests and assessed validity, intrarater reliability and interrater reliability. Three clinicians having varied levels of experience graded first-ray mobility on 15 subjects. A separate investigator measured dorsal mobility with a mechanical device. Both methods of testing were repeated to assess measurement reliability. Reliability was estimated by kappa (K) statistics. Spearman correlation assessed the relationship between mobility graded manually and dorsal mobility measured by device. Manual examination intrarater K values ranged from 0.50 to 0.85, and interrater agreement from 0.09 to 0.16. Manual grading was not related (r = -0.21) to the absolute measure of total dorsal mobility made by device. This brings into question the validity and reliability of manual estimates of first-ray mobility. PMID- 11934069 TI - Compression screw fixation for first metatarsal basal osteotomy. PMID- 11934070 TI - Chronic subluxation of the midtarsal joint of the foot: a case report. AB - Traumatic injuries to the midtarsal joint are quite uncommon and usually involve fractures of adjacent bones. Pure dislocations without fracture are very rare. A patient sustained a pure dislocation that was treated with a nonweight-bearing cast for six weeks, but she went on to present with chronic subluxation at the midtarsal joint. Imaging modalities were confusing because the pathology was seen only when the deformation was present throughout the imaging examination, not when it was reduced. Stress varus- valgus radiographs of the foot, not ankle, provided a definitive diagnosis. The patient was treated successful by arthrodesis of calcaneocuboid and talo-navicular joints (the midtarsal joint which was unstable) without subtalar fusion. PMID- 11934071 TI - Uremic tumoral calcinosis of the foot mimicking infection. AB - Uremic tumoral calcinosis is an uncommon, benign condition characterized by slow growing calcified periarticular soft tissue masses of varying size. We describe two patients with chronic renal failure on hemodialysis presenting uremic tumoral calcinosis, one in the fifth toe of the right foot and the other in the dorsum of the left foot between the first and second metatarsals. Excision of the calcic masses and parathyroidectomy were successfully performed in both patients. These cases are unusual in their rapid onset, mimicking acute infection. Differential diagnosis, radiological features and therapy are discussed. PMID- 11934072 TI - Intra-articular osteoid osteoma of the lower extremity: diagnostic problems. AB - We report five cases of intra-articular osteoid osteoma. Physical findings of osteoid osteoma vary with the site of the tumor. Juxta- and intra-articular osteoid osteomas present various atypical and nonspecific features. They are recorded both for their rarity and for the unusual clinical and roentgenographic findings that may delay diagnosis or induce misdiagnosis. Specialized imaging techniques may hasten diagnosis, but only an accurate clinical history, with a high index of suspicion, can allow for a proper diagnosis. When the diagnosis is suspected, we suggest that the most sensitive test is a "three-phase" technetium 99m bone-scan followed by computerized tomographic-scanning. Detailed evaluation of the suspected area, using thin sections is required to prevent misinterpretation, especially in a diagnosis involving first sprain of an ankle. The following cases of intra-articular osteoid osteoma illustrate the problems encountered in their diagnosis. PMID- 11934073 TI - Optimization study of doxorubicin liposomal preparations coated with laminin fragments. AB - Immunoliposomes, coated with two peptide sequences and loaded with doxorubicin, were prepared. The influence of different parameters in the sequential steps of liposomal preparations was studied as, for instance, lipid composition, size reduction methods, elimination of non-entrapped drug, and peptide coating sequence. Results were evaluated, such as entrapment efficiency, phospholipid/drug and phospholipid/peptide relationship, and size of final preparations. Effective size reduction was only achieved through probe sonication and the presence of peptides on the surface of liposomes, which does not modify, significantly, the final phospholipid/drug relationship, related to the initial values; however, they promoted a slight increase in the size of final preparations. Dialysis was the most suitable method to wash liposomes from reactants, drug and peptides, as well as being the cleanest process to avoid microbial contamination without significant dilution. Peptide coating yields were similar for liposomal compositions presenting free carboxyl groups on the surface. As determined by other authors, the presence of polyethylene glycol monomethoxy chains on the surface reduces the reactivity of NPGE carboxylic groups. PMID- 11934074 TI - Isolation and partial characterization of catalytic antibodies with oligonuclease activity from bovine colostrum. AB - Catalytic antibodies (abzymes) which hydrolyze RNA and DNA were isolated from bovine colostrum by sequential chromatography on Protein A Sepharose, denaturated DNA-cellulose, Mono Q, and gel permeation chromatography on Superose 12 at pH 2.3 after acidic shock. Metachromatic agar containing toluidine blue and yeast RNA was used to measure RNase activity. Electrophoresis in agarose showed DNase activity on plasmid DNA from Escherichia coli and DNA from calf thymus in fractions from all 4 purification steps. Gel permeation chromatography showed that the abzymes hydrolysed both a single-stranded polyadenylic acid (Poly A) and single-stranded polycitidylic acid (Poly C), while partially purified RNase from the colostrum hydrolysed Poly (C), but not Poly (A). Electrophoresis of purified abzymes under denaturing conditions showed protein bands of molecular mass corresponding to heavy and light chains of IgG. The abzymes immunoreacted with anti-bovine IgG. The RNase activity of the purified abzymes represented 0.022% of total RNase activity in the colostrum; acid shock and gel filtration at low pH reduced the specific RNase activity of abzymes 3.6-fold. The RNase activity of abzymes at pH 6.6 was reduced by 90% by heat treatment at 75 degrees C for 52 min. PMID- 11934075 TI - Synthesis of N-protected peptide alcohols catalyzed by subtilisin or alpha chymotrypsin in organic solvents. AB - A series of N-protected peptide alcohols were synthesized using amino alcohols with unprotected hydroxy groups as amino components by the catalysis of subtilisin or alpha-chymotrypsin in organic solvents. N-protected aromatic amino acid esters were more suitable as acyl donors for subtilisin. The influences of different N-protecting groups, organic solvents, and content of water on synthesis of N-protected peptide alcohols were systematically studied. PMID- 11934076 TI - Purification of turnip peroxidase and its kinetic properties. AB - Peroxidase from turnip roots was purified using metal affinity chromatography up to a specific activity of 337 units/mg protein with 3.02 RZ and 63.5% recovery. After purification, the enzyme showed 2-3 bands on sodium dodecyl sulphate polyacrylamide gel electrophoresis (SDS-PAGE). The molecular weight of the purified enzyme was found to be 37-39 kD with matrix assisted laser desorption ionization mass spectrometer (MALDI-MS). The enzyme showed maximum activity in phosphate buffer, pH 6.0, and lowest activity in borate buffer at the same pH. The Km of the enzyme was found to be 7.07 x 104 mM. Turnip peroxidase also contains an iron moiety which is found to be about 0.28%. The enzyme showed 50% inhibition of its specific activity with ethylene diamine tetraacetic acid (EDTA). PMID- 11934077 TI - Isolation and biochemical characterization of extracellular polymeric secretions (EPS) from modern soft marine stromatolites (Bahamas) and its inhibitory effect on CaCO3 precipitation. AB - Bahamian soft marine stromatolites consist of cyanobacterial biofilms and carbonate sand grains (ooids) embedded in their extracellular polymeric secretions (EPS). EPS were isolated from natural marine stromatolites and the laboratory cultured stromatolite forming cyanobacterium isolate Schizothix sp. Laboratory investigations were conducted to examine biochemical characteristics and the role of EPS in the inhibition of CaCO3 precipitation. EPS consisted of acid polysaccharides and proteins. SDS-PAGE and amino acid analysis suggested that EPS from both soft marine stromatolite and Schizothrix sp. mat contained small proteins (38 kD and 45 kD) enriched in aspartic acid and glutamic acid. Also, immuno blotting suggested that natural EPS contain high molecular weight acid polysaccharide (500 k) which may represent cross-linked products of laboratory cultured Schizothrix sp. acid polysaccharide (300 k). EPS from both soft marine stromatolite and laboratory cultured Schizothrix sp. inhibited CaCO3 precipitation in vitro, as determined using pH drift assays examining pH decrease which occur in response to CaCO3 precipitation. PH drift assays of enzymatically and chemically modified EPS isolated from soft marine stromatolite and laboratory cultured Schizothrix sp. indicated that both uronic acids and protein fractions may be involved in the inhibition of CaCO3 precipitation. PMID- 11934078 TI - HPLC preparation of fish waste hydrolysate fractions. Effect on guinea pig ileum and ACE activity. AB - The effect of RP-HPLC-purified fractions of fish waste hydrolysates issued from three fish industries was tested on guinea pig ileum in order to examine the presence of opioid molecules. The evaluation of anti-hypertensive activities of whole hydrolysates and fractions were also tested, monitoring the ability of the fraction to inhibit the activity of angiotensin I-converting enzyme involved in hypertension regulation. Sardine autolysate and cod head hydrolysate powder (50 microg) were able to inhibit near 30% of ACE activity, whereas 50 microg of shrimp hydrolysate allows the inhibition of 57% of ACE activity. HPLC fractionation of cod head hydrolysate and sardine autolysate was necessary to evidence biological activity, whereas HPLC separation of shrimp hydrolysate exhibited low biological activity fractions. Further studies are necessary to characterise bioactive molecules from cod head alcalase hydrolysate and from sardine autolysate. PMID- 11934079 TI - Synthesis of two neuromedin U (NMU) analogues and their comparative effect of reducing food intake in rats. AB - To examine the roles of aromatic rings, Tyr residues at positions 1 and 5 and Phe residues at positions 16, 17, and 19 of rat neuromedin U-23 (NMU-23) (Tyr-Lys-Val Asn-Glu-Tyr-Gln-Gly-Pro-Val-Ala-Pro-Ser-Gly-Gly-Phe-Phe-Leu-Phe-Arg-Pro-Arg-Asn NH2) for reducing food intake activity in male Wistar rats, two NMU-23 analogues, [Phe(4F)16,17,19] NMU-23 and [Tyr(Me)1,6]NMU-23, were synthesized by Fmoc strategy of manual solid-phase method. The synthetic NMU-23 showed reducing effect on food intake in rats. [Phe(4F)16,17,19]NMU-23 exhibited higher reducing food in take effect than that of NMU-23. On the contrary, [Tyr(- Me)1,6]NMU-23 showed no reducing effect on food intake in rats than that of NMU-23. PMID- 11934080 TI - Brief treatment for young substance abusers: a pilot study in an addiction treatment setting. AB - The present study evaluated a motivationally based, 4-session outpatient intervention for young substance abusers presenting for addiction treatment. Follow-up interviews were conducted 6 months after assessment on (a) clients who sought additional help (n = 22) and (b) clients who did not seek additional help (n = 28). Results indicated that (a) participating in an assessment and brief intervention was associated with reduced use and consequences and increased confidence in high-risk situations up to 6 months after entry into the program, and (b) clients who participated in additional treatment showed less of a decrease in substance-related consequences. These findings suggest that brief interventions can serve as either a stand-alone intervention for individuals who need short-term assistance or a first-step intervention for those with more specialized or long-term needs. PMID- 11934081 TI - Workplace absenteeism and alcohol use: a sequential analysis. AB - This investigation examined the day-to-day relationship between alcohol use and workplace absenteeism among a sample of participants (N = 280) employed in 1 of 3 large companies located in the northeastern U.S. With a semistructured interview, information was collected from employees about specific days of drinking during a 1-month period and marked on a calendar. Data about employees' absences during the same target time period were collected from the companies' human resource departments and were also marked on a calendar. A significant relationship was found between alcohol use and workplace absences; workers were roughly 2 times more likely to be absent from work the day after alcohol was consumed. PMID- 11934082 TI - Predictors of quitting and dropout among women in a clinic-based smoking cessation program. AB - Most cessation studies assume that dropouts are smokers. Instead, the authors analyzed these outcomes separately using multinomial regression to model the relative risk of quitting versus continued smoking and dropping out. Female (N = 281) smokers were randomly assigned to a 12-week smoking cessation program plus either a 3-times-per-week exercise program or a contact control wellness program. Higher body mass index and longer prior quit attempts predicted cessation. Self efficacy was associated with a lower likelihood of dropout. Greater nicotine dependence and lower education predicted continued smoking or dropout versus quitting among exercisers. Patterns of smoking, dropping out, and quitting between Weeks 5 and 12 were different between exercisers and controls. Dropouts should be considered as a separate category from smokers. PMID- 11934083 TI - Measuring substance-free and substance-related reinforcement in the natural environment. AB - The present study sought to provide further evidence for the validity of a modified version of the Pleasant Events Schedule (PES; D. J. MacPhillamy & P. M. Lewinsohn, 1982) designed to measure substance-free and substance-related reinforcement. A sample of 134 young adults completed the modified PES along with measures of substance use and quality of life. The results extend previous research on the modified PES in 3 ways: (a) Information regarding the relationships between substance-related reinforcement and substance use are expanded to include substance-use frequency, quantity, and related negative consequences; (b) relationships between substance-free reinforcement and non substance-related variables are established; and (c) the distinctiveness of the substance-free and substance-related reinforcement scores is demonstrated. The utility of reinforcement surveys in the study of substance use is discussed, with special emphasis placed on possible treatment implications. PMID- 11934084 TI - A prospective study of youth gambling behaviors. AB - Little is known about the course and outcomes of adolescent gambling. This prospective study describes findings from a 3-wave (Time 1 [T1], Time 2 [T2], and Time 3 [T3]) assessment of gambling behaviors among youth (N = 305). Stable rates of any gambling and regular gambling (weekly or daily) were observed across T1, T2, and T3. The rate of at-risk gambling significantly increased at T3 (young adulthood), whereas the rate of problem gambling remained stable over time. Several adolescent risk factors were associated with either T3 at-risk or problem gambling, many of which are risk factors for adolescent substance abuse. Findings suggest that important to the origins of young adult gambling problems are risk factors associated with the problem behavior syndrome of adolescence. PMID- 11934085 TI - Individual differences in latent neuropsychological abilities at addictions treatment entry. AB - The latent structure of neuropsychological abilities and risk factors for impairment were examined in 197 persons entering addictions treatment. Confirmatory factor analysis yielded 4 factors: Executive, Memory, Verbal, and Processing Speed. The measurement model was consistent with evidence that neuropsychological test performance is factorially complex and supported by multiple brain regions. Path analyses showed that risk factors explained 34%-57% of the true variance in abilities. Age, education, and medical status had the most generalized and robust associations with abilities. Drug use disorder diagnoses, childhood behavior problems, familial alcoholism, and psychopathology were also significantly related to specific latent abilities. Knowledge of neuropsychological impairment may be clinically useful, and selected risk factors may help treatment providers decide which clients should receive formal neuropsychological assessment. PMID- 11934086 TI - Alcohol's effect on aggression identification: a two-channel theory. AB - Identification of ambiguous behaviors may be affected by alcohol first by the activation of associated mental representations and second by an increase in the imbiber's motivation of need for closure (NFC; A. Kruglanski, 1989), because cognitive effort is increased for epistemic activities. Combined, these effects should increase correspondence between mental representations of alcohol and the identification of others' behaviors. Three studies were conducted to test this hypothesis. The results were consistent with this hypothesis: Participants who associated alcohol with amiable concepts perceived less aggressive intent when blood alcohol concentrations (BACs) were high versus low. Alternatively, those who associated alcohol with aggressive concepts perceived the same or more aggressive intent when BACs were high versus low. Priming alcohol concepts and trait-level NFC were also sufficient to replicate these effects. PMID- 11934087 TI - Test-retest reliability of alcohol measures: is there a difference between internet-based assessment and traditional methods? AB - This study compared Web-based assessment techniques with traditional paper-based methods of commonly used measures of alcohol use. Test-retest reliabilities were obtained, and tests of validity were conducted. A total of 255 participants were randomly assigned to 1 of 3 conditions: paper-based (P&P), Web-based (Web), or Web-based with interruption (Web-I). Follow-up assessments 1 week later indicated reliabilities ranging from .59 to .93 within all measures and across all assessment methods. Significantly high test-retest reliability coefficients support the use of these measures for research and clinical applications. Furthermore, no significant differences were found between assessment techniques, suggesting that Web-based methods are a suitable alternative to more traditional methods. This cost-efficient alternative has the advantage of minimizing data collection and entry errors while increasing survey accessibility. PMID- 11934088 TI - Sex-related alcohol expectancies predict sexual risk behavior among severely and persistently mentally ill adults. AB - Three hundred three adults (57% male, average age 42 years) with severe and persistent mental illness receiving treatment at community mental health clinics completed a survey, which included B. C. Leigh's (1990) sex-related alcohol expectancy scale and measures of alcohol use and sexual risk behavior. Hierarchical logistic regression analyses, controlling for drinking behavior, revealed that participants with stronger expectancies that drinking would lead to enhanced sexual experience were more likely to have drank prior to intercourse and that, among participants who drank prior to intercourse, those with stronger expectancies that alcohol would lead to riskier sexual behavior were more likely to have engaged in sexual risk behavior. Implications for preventing HIV infection among people with severe mental illness are discussed. PMID- 11934089 TI - Depressive symptoms, stress, and weight concerns among African American and European American low-income female smokers. AB - The relationships between perceived stress, depressive symptoms, concern about weight gain and smoking dependence were examined among 83 European American and 175 African American female smokers bringing children to pediatric clinics serving a low-income population. Among African American women, but not European American women, greater stress and more depressive symptoms predicted greater smoking dependence, and less concern about weight gain predicted greater smoking dependence. Multivariate analyses confirmed the bivariate relationships among stress, depressive symptoms, and smoking dependence among African American women but reduced the relationship between weight concern and smoking dependence. The stronger relationships among stress, depressive symptoms, and smoking dependence among African American women may be indicative of smoking patterns more associated with affect regulation than are the smoking patterns of European American women. PMID- 11934090 TI - Risk and vulnerability for marijuana use problems: the role of affect dysregulation. AB - This study examined the moderating effects of impulsivity and affect lability on relations between marijuana use frequency and use consequences. From a sample of 592 undergraduates, 300 marijuana users completed a survey that assessed marijuana problems and the hypothesized risk and vulnerability factors. Affective variables were significantly associated with increased marijuana problems above and beyond the effects of gender and lifetime use frequency. A hypothesized vulnerability mechanism, whereby impulsivity strengthened the relationship between use frequency and marijuana problems, was observed. The findings support the hypothesis that affect dysregulation enhances risk for marijuana problems among young adults who use marijuana. PMID- 11934091 TI - Disordered gambling among university-based medical and dental patients: a focus on Internet gambling. AB - The authors evaluated gambling behaviors, including Internet gambling, among patients seeking free or reduced-cost dental or health care. Three hundred eighty nine patients at university health clinics completed a questionnaire that included the South Oaks Gambling Screen (SOGS; H. R. Lesieur & S. Blume, 1987). All respondents had gambled in their lifetimes, with 70% gambling in the past 2 months. On the basis of SOGS scores, 10.6% were problem gamblers, and 15.4% were pathological gamblers. The most common forms of gambling were lottery, slot machines, and scratch tickets. Internet gambling was reported by 8.1% of participants. Compared to non-Internet gamblers, Internet gamblers were more likely to be younger, non-Caucasian, and have higher SOGS scores. This study is among the first to evaluate the prevalence of Internet gambling and suggests that people who gamble on the Internet are likely to have a gambling problem. Results also illuminate the need to screen patients seeking health care services for gambling problems. PMID- 11934092 TI - Three-dimensional sonographic measurement of liver volume in the small-for gestational-age fetus. AB - OBJECTIVE: To assess the growth of the fetal liver in normal pregnancies and to evaluate the ability of fetal liver volume measurement for prediction of small for-gestational-age fetuses. METHODS: Three-dimensional sonographic examinations were performed on 14 appropriate-for-gestational-age and 10 small-for-gestational age fetuses. Liver volume and liver length were measured every 2 weeks after 20 weeks' menstrual age until delivery. RESULTS: A curvilinear relationship was found between the menstrual age and liver volume (R2 = 88.4%; P < .0001), and a normal range of liver volume measurement for estimating the growth of the fetal liver during normal pregnancy was generated. Liver length was normal in 7 of 10 small-for-gestational-age fetuses, whereas liver volume values in all small-for gestational-age fetuses were below normal ranges in the mid to late third trimester. CONCLUSIONS: Our findings suggest that liver volume may be a useful measurement for diagnosing small-for-gestational-age fetuses in the mid to late third trimester but that liver length may not be predictive. Further studies involving a larger sample size would be needed to confirm this suggestion. PMID- 11934093 TI - Effects of oophorectomy and hormone replacement therapy on the pulsatility indices of hepatic and renal arteries. AB - OBJECTIVE: To investigate the flow velocity waveform changes of the hepatic and renal arteries in women with surgical menopause who received hormone replacement therapy versus those who did not. METHODS: Eighty women who had undergone surgical menopause were divided into 2 groups. The first group (n = 38) consisted of patients who did not receive estrogen treatments after surgery; patients in the second group (n = 42) did receive treatments. The hepatic and renal arteries of patients in both groups were examined by duplex Doppler ultrasonography before the commencement of hormone replacement therapy and after 2 years of treatment, and the pulsatility indices were calculated. RESULTS: No significant differences were detected in the renal and hepatic artery pulsatility indices of patients in the estrogen treatment group (group 2) before and after total abdominal hysterectomy and bilateral salpingo-oophorectomy (P > .05). No significant differences in preoperative and postoperative hepatic arterial pulsatility indices were detected among patients in group 1 (P > .05). Renal artery pulsatility indices measured before and after total abdominal hysterectomy and bilateral salpingooophorectomy did show a statistically significant difference in group 1 (P < .001). In addition, a statistically significant difference was detected before and after surgery in both groups when pulsatility indices were measured at the second-year control dose. CONCLUSIONS: Hepatic arterial pulsatility indices are not affected in postmenopausal women, but renal artery pulsatility indices rise to some extent in women not receiving hormone replacement therapy. PMID- 11934094 TI - Ultrasonographic findings correspond to clinical, endoscopic, and histologic findings in inflammatory bowel disease and other enterocolitides. AB - OBJECTIVE: To compare results obtained by abdominal ultrasonography with clinical findings, including endoscopic and histologic findings, to evaluate the location and activity of inflammatory bowel disease, including disease controls in children. METHODS: Ninety-two ultrasonographic scans and 41 colonoscopic examinations with biopsies were performed in 78 patients (1 month to 17.8 years of age) with Crohn's disease (n = 26), ulcerative colitis (n = 21), inflammatory bowel disease of indeterminate type (n = 2), and disease controls (other intestinal disorders, including infectious and ischemic lesions; n = 29). Laboratory parameters for inflammatory bowel disease were determined, and disease activity was assessed by a combination of clinical and laboratory data. Bowel wall thickness and echo texture were recorded in a standardized way by ultrasonography and compared with endoscopic and histologic findings in a segment by-segment comparison. RESULTS: Sensitivity and specificity of ultrasonography in detecting patients with severe macroscopic lesions depicted on endoscopy were 77% and 83%, respectively. Sensitivity and specificity of ultrasonography in detecting patients with severe histologic inflammation were 75% and 82%. There was a statistically significant correlation between maximal bowel wall thickness and disease activity score (P < .01). CONCLUSIONS: Abdominal ultrasonography may be helpful in evaluating the location, severity, and inflammatory activity of inflammatory bowel disease in children and young adults. PMID- 11934095 TI - Diagnosis of posterior enterocele: comparison of rectal ultrasonography with intraoperative diagnosis. AB - OBJECTIVE: To describe a new ultrasonographic technique and to compare it with intraoperative findings in women with a suspected enterocele. METHODS: An observational study was conducted in the Department of Gynecology of a medium sized teaching hospital. Twenty-nine patients with clinical and ultrasonographic diagnoses of an enterocele were reviewed. All patients had vaginal repair surgery, including a posterior repair in which the existence of an enterocele was confirmed or denied. RESULTS: In 27 of the 29 patients, the enterocele was diagnosed as such during surgery and handled accordingly. The 2 enteroceles that were not confirmed were smaller types. CONCLUSIONS: Rectal ultrasonographic findings were in good accordance with intraoperative anatomic characteristics. Rectal ultrasonography has the capability of diagnosing small, occult enteroceles. This can have important clinical implications in the prophylactic use of additional enterocele repair during hysterectomy or prolapse surgery. PMID- 11934096 TI - Assessment of therapeutic response in hepatocellular carcinoma treated with percutaneous radio frequency ablation: comparison of multiphase helical computed tomography and power Doppler ultrasonography with a microbubble contrast agent. AB - OBJECTIVE: To compare the results of multiphase helical computed tomography and power Doppler ultrasonography with a microbubble contrast agent in the assessment of the therapeutic response to radio frequency ablation in hepatocellular carcinoma. METHODS: In 66 patients with 73 nodular hepatocellular carcinomas ranging from 1.0 to 4.0 cm (mean, 2.6 cm) in diameter, contrast-enhanced power Doppler ultrasonography was performed after intravenous bolus injection of a galactose-based microbubble contrast agent before and after radio frequency ablation. The results of the studies were compared with the findings of follow-up 3-phase helical computed tomography. All patients were regularly followed up with computed tomography for more than 1 year (range, 13-19 months). RESULTS: In 8 (11%) of 73 hepatocellular carcinomas, immediate follow-up computed tomography obtained within 2 hours after radio frequency ablation showed focal enhancing portions within the treated lesions, suggesting residual non-necrotic tumors. All 8 of these tumors had intratumoral flow signals on contrast-enhanced power Doppler ultrasonography. The diagnostic agreement between computed tomography and contrast-enhanced power Doppler ultrasonography was achieved in 100%. Among the remaining 65 hepatocellular carcinomas with the absence of residual tumors at both immediate follow-up computed tomography and contrast-enhanced power Doppler ultrasonography, subsequent follow-up computed tomography showed local regrowth at the margins of 10 lesions (15%). CONCLUSIONS: The results of contrast-enhanced power Doppler ultrasonography closely correlated with those of immediate follow up computed tomography for detecting residual tumors in hepatocellular carcinomas treated with radio frequency ablation. Both techniques, however, showed a limitation in detecting small or microscopic residual tumors and in predicting local regrowth in the treated lesions. PMID- 11934097 TI - Color Doppler sonography in the evaluation of superficial lymphomatous lymph nodes. AB - OBJECTIVE: We performed a retrospective study to document the sonographic and color Doppler characteristics of lymphomatous superficial lymph nodes. METHODS: We selected 130 individuals who underwent sonography, color Doppler imaging, fine needle aspiration biopsy, and surgical removal of the nodes with the final diagnosis of lymphoma (87) and chronic adenitis (43). During sonography, for each node we considered the longitudinal and axial diameters, long/short axis ratio, visibility of the hilum, and the internal echogenicity of the node. During the color and power Doppler examination, we classified the nodes into 3 patterns: type I, "hilar normal"; type II, "hilar activated"; and type III, "peripheral." RESULTS: Sonographic evidence was not significant. With color Doppler sonography, 97% of nodes affected by non-Hodgkin lymphoma, 94% of nodes affected by Hodgkin lymphoma, and 100% of non-neoplastic nodes showed hilar vascularity. Type I seemed more frequently associated with inflammation, and type II was more frequently associated with lymphoma. CONCLUSIONS: The presence of peripheric subcapsular vessels, which is typical of metastasis, is definitely rare in lymphoma (with the possible exception of the uncommon subtypes of high-grade lymphomas). The differential diagnosis between lymphoma and lymphadenitis is frequently impossible on the basis of sonographic and color Doppler patterns alone; therefore, clinical evaluation and biopsy are generally mandatory. PMID- 11934098 TI - Sonographic findings of the hepatobiliary-pancreatic system in adult patients with cystic fibrosis. AB - OBJECTIVE: Sonography of the liver, biliary system, and pancreas in adult patients with cystic fibrosis is by far less systematically documented than in pediatric patients with cystic fibrosis. In this prospective study, duplex sonographic findings of the liver, biliary system, and pancreas in adult patients with cystic fibrosis were compared with those of healthy control subjects. METHODS: Seventy-two consecutive patients with cystic fibrosis and 60 healthy control subjects were examined by high-resolution sonography. The incidence of perihepatic lymphadenopathy, the hepatic echo pattern, the detection rate of liver tumors, the flow patterns in the hepatic and portal veins, and pathologic gallbladder and pancreas findings were recorded. Additionally, cholestasis indicating enzyme levels (gamma-glutamyl transpeptidase and alkaline phosphatase), liver function test results (alanine aminotransferase and aspartate aminotransferase levels), and amylase and lipase levels were recorded as well. RESULTS: Patients with cystic fibrosis, when compared with healthy subjects on sonographic examination, had a higher incidence of microgallbladder (25% versus 0%) and cystic lesions of the pancreas (18% versus 0%). The number of abnormal echo patterns of the liver was increased (46% versus 15%), with a higher incidence of a nontriphasic flow pattern in the right hepatic vein. The differences proved to be statistically significant (P < .05). CONCLUSIONS: Typical sonographic findings in adult patients with cystic fibrosis are a microgallbladder and small cystic lesions of the pancreas. Pathologic findings of the liver can be shown by B-mode and duplex sonography, but the resulting patterns are less characteristic. PMID- 11934099 TI - Observation on the integrity of the blood-brain barrier after microbubble destruction by diagnostic transcranial color-coded sonography. AB - OBJECTIVE: To investigate alteration of the blood-brain barrier from ultrasonic contrast agent destruction by diagnostic transcranial color-coded sonography using gadolinium-enhanced magnetic resonance imaging. METHODS: Healthy male volunteers received 10 mL (400 mg/dL) of Levovist (SH U 508A; Schering AG, Berlin, Germany; n = 6) or 3 mL of Optison (FS069; Mallinckrodt Inc, St Louis, MO; n = 4) followed by 0.3 mmol/kg magnetic resonance imaging contrast agent (Magnevist; Schering) intravenously. Then transcranial color-coded sonography was performed with a conventional color duplex sonographic system, which insonated the brain in a slightly angulated axial plane with temporal average intensity of less than 700 mW/cm2 or acoustic pressure amplitude of less than 2.69 MPa, attenuated by the temporal bone. Before, immediately after, and 2 hours after insonation, T1-weighted axial magnetic resonance imaging was performed. All magnetic resonance images were individually assessed, and T1 signal intensities were measured in 2 regions of interest in both hemispheres at the 3 time points. RESULTS: No focal contrast enhancement or damage to the brain and no significant difference between T1 signal intensities in the right and left brain regions could be detected during early or late phases when either ultrasonic contrast agent was used. CONCLUSIONS: This bioeffects study gives further evidence of the safety of ultrasonic destruction of Levovist and Optison microbubbles by diagnostic transcranial color-coded sonography. However, more subtle local effects may have been missed by gadolinium-enhanced magnetic resonance imaging. Studies on diagnostic contrast-enhanced transcranial color-coded sonography as well as microbubble-based drug delivery strategies should consider ultrasonic contrast agent microbubble characteristics and concentration as well as ultrasound transmission power levels. PMID- 11934100 TI - Sonography in renovascular hypertension. AB - OBJECTIVE: To familiarize practitioners with different sonographic manifestations of renal artery compromise and the sonographic techniques for renal artery imaging. METHODS: Approximately 1500 examinations evaluating for renal artery disease are performed in our vascular laboratory every year. Most of the patients have the symptoms of hypertension (possibly related to renovascular etiology) and renal insufficiency. From our cumulative experience, the optimal scanning techniques are defined for each renal artery, for extrarenal versus intrarenal vascular evaluation, and for patients with different body habitus. We have also tabulated our technical success rate. Cases with sonographic evidence of renal artery compromise are identified. The validity, sensitivity, and specificity of different parameters are examined. RESULTS: We achieve an approximately 75% to 80% success rate in obtaining technically adequate studies. We have not found the tardus-parvus waveform evaluation to be as valuable as direct interrogation of the renal artery. CONCLUSIONS: Duplex/color Doppler sonography serves a vital role in the diagnosis of renal artery stenosis and occlusion; it has an excellent correlation with contrast-enhanced angiography. It is also used for intraoperative or postrevascularization surveillance to show evidence of recurring stenosis, thrombosis, and other complications. PMID- 11934101 TI - Three-dimensional ultrasonography using the minimum transparent mode in obstructive biliary diseases: early experience. AB - OBJECTIVE: To assess the feasibility of three-dimensional ultrasonography using the minimum transparent mode in patients with obstructive biliary disease. METHODS: Fourteen patients with obstructive jaundice underwent three-dimensional ultrasonography after conventional two-dimensional ultrasonography before interventional procedures. Three-dimensional images reconstructed by the minimum transparent mode were evaluated and compared with direct cholangiography. The minimum transparent mode images were compared with the two-dimensional ultrasonographic images to determine whether additional information was obtained. RESULTS: The level of obstruction was correctly depicted in all patients, and a dilated common bile duct, common hepatic duct, gallbladder, and main intrahepatic ducts were well visualized on minimum transparent mode images. The findings on minimum transparent mode images were well correlated with those on cholangiography; however, the perspective of the whole biliary tree on minimum transparent mode images was inferior to that on cholangiography in all cases. The overall image quality of minimum transparent mode images was poor in 6 patients, fair in 3, good in 4, and excellent in 1. The biliary anatomy was more objectively visualized on the minimum transparent mode images than conventional two-dimensional ultrasonographic images. CONCLUSIONS: Three-dimensional ultrasonography using the minimum transparent mode appears to be clinically feasible. The biliary anatomy was more objectively visualized on the minimum transparent mode images than conventional two-dimensional ultrasonographic images. Further technical development is needed to improve imaging resolution. PMID- 11934103 TI - Transvaginal sonography in the treatment of a rare case of total urethral stenosis with a vesicovaginal fistula. PMID- 11934102 TI - Persistent median artery in the carpal tunnel: color Doppler ultrasonographic findings. AB - OBJECTIVE: To describe the ultrasonographic and color Doppler ultrasonographic findings in 2 patients with carpal tunnel syndrome associated with a persistent median artery and to report the frequency of this anatomic variation in healthy volunteers. METHODS: Two patients with the clinical appearance of carpal tunnel syndrome and 100 wrists and distal forearms of 50 asymptomatic volunteers were examined with ultrasonography and color Doppler ultrasonography. The frequency and size of a persistent median artery and its relationship to median nerve anatomy in the carpal tunnel were evaluated. RESULTS: A large persistent median artery of 3 mm in diameter was found in the affected hands in both patients with carpal tunnel syndrome. Findings were confirmed at surgery. Among the asymptomatic volunteers, a persistent median artery could be found in 13 (26%, 10 [20%] unilateral and 3 [6%] bilateral), with a mean diameter of 1.1 mm (range, 0.5-1.7 mm). In 10 (63%) of 16 hands, the persistent median artery was associated with high division of the median nerve or a bifid nerve configuration in the carpal tunnel. CONCLUSIONS: A persistent median artery is a common condition in healthy individuals and in most cases is related to median nerve variations such as high division or a bifid nerve. Because a persistent median artery has a superficial course close to the transverse carpal ligament, preoperative diagnosis of this anatomic variation may be of clinical importance. PMID- 11934104 TI - Transvaginal sonographic and power Doppler diagnosis of endometrial lymphoma. PMID- 11934105 TI - Large mesenteric lipoma ultrasonographically mimicking a mature cystic teratoma during pregnancy. PMID- 11934106 TI - A possible new ancillary sign for diagnosing midgut volvulus: the truncated superior mesenteric artery. PMID- 11934107 TI - First-trimester nuchal translucency screening. PMID- 11934108 TI - First-trimester nuchal translucency screening. PMID- 11934109 TI - Stress-70 as indicator of heavy metals accumulation in blue mussel Mytilus edulis. AB - The induction of stress-70 (Hsp70) in the blue mussels Mytilus edulis from southern Baltic after exposure to cadmium, lead and copper, administrated individually and/or as a mixture, was investigated by means of standard Western blotting using a clone 3A3 monoclonal antibody. The general course of the stress 70 induction was found to be similar for all tested metals. It could be characterised by three main features: (1) The induction of stress-70 proteins was only slightly increased at low metal concentration. (2) Strong induction of stress-70 occurred at high concentration of metals. Combination of metals (Cd+Cu) was found to increase the stress-70 level more intensely than related concentrations of singularly applied metals. (3) Five weeks of long depuration had no effect on stress-70 levels. Experiments revealed different induction of stress-70 as a response to cadmium, lead and copper exposure and confirm that the applied technique might serve as a versatile tool to assess invertebrate stress reaction to environmentally relevant heavy metals. PMID- 11934110 TI - Leachability of elements from sub-bituminous coal fly ash from India. AB - Environmental concerns regarding the potential contamination of soil, surface and ground water due to the presence of soluble metal species in the ash pond leachate is of great importance. Serial batch leaching was carried out simulating the rainwater condition of the study area to understand the behaviour of elements during leaching. The leachates were analysed for the elements Al, Ca, K, Mg, Na, P, S, Si, As, Ba, Fe, Mn, Mo, Ti, V, Pb, Zn, Co, Cr, Cu, Ni and Cd by inductively coupled plasma optical emission spectrometer (ICP-OES). It was found that Cd, Co, Cr and Ni did not leach from the ash while Cu and Pb concentrations were insignificant in the leachate regardless of liquid to solid (L/S) ratio. Most of the elements showed maximum concentrations at lower L/S ratio and then decreased with increasing L/S. The total cumulative concentrations of As, Mn and Mo were found to be higher than the World Health Organization (WHO) recommended values for drinking water while the concentrations of Fe, Mn and As exceeded the maximum allowable concentrations prescribed by the United States Environmental Protection Agency (USEPA). The pre and the post leached ash samples were analysed for morphology, specific surface area and mineralogical changes. Analysis of post leached fly ash indicated changes in the specific surface area and morphology but no change in mineralogy. PMID- 11934111 TI - Estimating the total exposure to air pollutants for different population age groups in Hong Kong. AB - Given that Hong Kong is one of the most densely populated cities in the world, the exposure of the Hong Kong people is one of the interesting research areas. In this study, an indirect approach was used to estimate the exposure to nitrogen dioxide (NO2), respiratory dust (PM10) and carbon monoxide (CO) pollutants experienced by different age groups of people in Hong Kong. The average concentrations of the 20 major microenvironments obtained from our measurement survey data, together with the people activity pattern data obtained from 7-day recall questionnaires, were used to predict frequency distributions to exposure assessment. Our results showed that Hong Kong people spent more than 86% of their time indoors. Homes were shown to be the one of the major exposure sites to NO2, CO and PM10 for all age groups. Our results also indicate that the 24-h NO2 exposure for individuals, irrespective of age, spending more than 2 h in commuting daily, was observed to be exceeding the 24-h NO2 exposure standards. This study was one of the pioneering studies with valuable contribution for modeling the estimates of exposures to NO2, PM10 and CO of different age groups in Hong Kong. PMID- 11934112 TI - Delineation of groundwater contamination around an ash pond: geochemical and GIS approach. AB - The study has investigated the levels of metal contamination in groundwater due to particulate matter fallout and leaching from ash pond and assigned contamination indices for the adjacent localities around an ash disposal site with application of geographic information systems (GIS). Fe, Ba, Cu, Mn, S, Pb, V, and Zn were found to be the major contaminants in groundwater. Enrichment factors (EF) of these elements with respect to the United States Environmental Protection Agency (USEPA) maximum contaminant levels show high values for Mn, Fe, and Pb in groundwater. The zone of attenuation for Ba, Fe, Cu, Mn, S, and Zn in groundwater is about 600-900 m from the ash pond, while Pb did not show any significant attenuation even at a distance of 1200 m. Tube wells around Rankasingha and Kukurhanga villages are most contaminated whereas open wells of Lachhmanpur, Kaniapada, and Kurudul villages showed higher degrees of contamination. PMID- 11934113 TI - Radioactive impact in sediments from an estuarine system affected by industrial wastes releases. AB - A big fertilizer industrial complex and a vast extension of phosphogypsum piles (12 km2), sited in the estuary formed by the Odiel and Tinto river mouths (southwest of Spain), are producing an unambiguous radioactive impact in their surrounding aquatic environment through radionuclides from the U-series. The levels and distribution of radionuclides in sediments from this estuarine system have been determined. The analyses of radionuclide concentrations and activity ratios have provided us with an interesting information to evaluate the extension, degree and routes of the radioactive impact, as well as for the knowledge of the different pathways followed for the radioactive contamination to disturb this natural system. The obtained results indicate that the main pathway of radioactive contamination of the estuary is through the dissolution in its waters of the radionuclides released by the industrial activities and their later fixation on the particulate materials. Tidal activity also plays an important role in the transport and homogenization along the estuary of the radioactivity released from the fertilizer plants. PMID- 11934114 TI - The origin of faeces by means of biomarker detection. PMID- 11934115 TI - Odour from municipal solid waste (MSW) landfills: a study on the analysis of perception. AB - The objective of this work was to develop a relationship between odour intensity and odour concentration by using data collected from various sensitive areas of the municipal solid waste (MSW) landfill site. A number of well-known psychophysical models (e.g., Weber-Fechner law, Steven's power law, Beidler's and Laffort's models) have been discussed that can successfully relate the perceived intensity with the odour concentration. Respective parameters for each of the models were estimated by the nonlinear Levenburg-Marquardt parameter estimation method. The overall performance of the model was tested statistically against sets of data from the olfactometry analysis. The model based on the Weber-Fechner law was ranked 1 in case of five out of nine samples and it has been found more representative of the less intense odour samples. The model based on Laffort's equation has represented the intensity-concentration relationship better with extremely low uncertainties on both parameters k1 and k2 for comparatively more intense odour samples. PMID- 11934116 TI - An approach to comparative assessments of potential health risks from exposure to radionuclides and hazardous chemicals. AB - The need to compare potential health risks to the public associated with different activities that can result in releases of hazardous substances to the environment is becoming increasingly important in decision-making. In making such comparisons, it is desirable to use equivalent indicators of potential health risks for radionuclides, chemical carcinogens, and noncarcinogenic hazardous chemicals. Current approaches to risk assessment that were developed for purposes of protecting human health do not provide equivalent indicators of potential risks from exposure to radionuclides and hazardous chemicals. Comparisons of environmental concentrations or calculated exposures or risks with standards for protection of public health also do not provide equivalent indicators of potential risks. We propose a simple approach to comparative risk assessments in which calculated exposures to any hazardous substances are expressed relative to no-observed-effect levels (NOELs) or, preferably, lower confidence limits of benchmark doses (BMDLs) in humans. This approach provides an equivalent, science based indicator of the relative risks posed by different exposures to any hazardous substances. PMID- 11934117 TI - The use of the oxidative stress responses as biomarkers in Nile tilapia (Oreochromis niloticus) exposed to in vivo cadmium contamination. AB - Water contaminants have a high potential risk for the health of populations. Protection from toxic effects of environmental water pollutants primarily involves considering the mechanism of low level toxicity and likely biological effects in organisms who live in these polluted waters. The biomarkers assessment of oxidative stress and metabolic alterations to cadmium exposure were evaluated in Nile tilapia, Oreochromis niloticus. The fish were exposed to 0.35, 0.75, 1.5, and 3.0 mg/l concentrations of Cd2+ (CdCl2) in water for 60 days. Fish that survived cadmium exposure showed a metabolic shift and a compensatory development for maintenance of the body weight gain. We observed a decreased glycogen content and decreased glucose uptake in white muscle. Lactate dehydrogenase (LDH) and creatine phosphokinase (CK) activities were also decreased, indicating that the glycolytic capacity was decreased in this tissue. No alterations were observed in total protein content in white muscle due to cadmium exposure suggesting a metabolic shift of carbohydrate metabolism to maintenance of the muscle protein reserve. There was an increase in glucose uptake, CK increased activity, and a clear increase of LDH activity in red muscle of fish with cadmium exposure. Since no alterations were observed in lipoperoxide concentration, while antioxidant enzymes glutathione peroxidase (GSH-Px) and superoxide dismutase (SOD) were changed in the liver and the red and white muscle of fish with cadmium exposure, we can conclude that oxygen free radicals are produced as a mediator of cadmium toxicity. Resistance development is related with increased activities of antioxidant enzymes, which were important in the protection against cadmium damage, inhibiting lipoperoxide formation. PMID- 11934118 TI - Effect of soil texture on surfactant-based remediation of hydrophobic organic contaminated soil. AB - Surfactants may be used in remediation of subsoil and aquifer contaminated with hydrophobic compounds. The objectives of this study were to examine the effect of soil texture on hydrophobic organic contaminant (HOC; toluene, or 1,2,4 trichlorobenzene [TCB]) removal from six soils and to evaluate the optimal composition of soil texture for maximum HOC removal using aqueous surfactant solution. Selected surfactants were 4% (vol/vol) sodium diphenyl oxide disulfonate (DOSL) and 4% (wt/vol) sodium lauryl sulfate (LS). Toluene and TCB were selected as the lighter-than-water nonaqueous phase liquid (LNAPL) and denser-than-water nonaqueous phase liquid (DNAPL) model substances, respectively. Soil types used for this study were Ottawa sand and five Iowa soils (Fruitfield, Keomah, Crippin, Webster, and Galvar). The greatest recovery of toluene and TCB in batch tests was 73% and 84%, respectively, which was obtained with DOSL surfactant in Ottawa sand. The toluene removal of 95% in column tests has been achieved in the Ottawa sand and three Iowa soils (Fruitfield, Keomah, Crippin) with DOSL after effluent volume of 3750 ml (about 32 pore volume) passed. TCB removal of 98% in column tests has been achieved in Ottawa sand and three Iowa soils (Fruitfield, Keomah, Crippin) with DOSL after effluent volume of 2500 ml (about 21 pore volume) passed. These results were related with soil texture (clay content 30%), clay mineralogy (kaolinite and smectite), as a function of transported pore volume. PMID- 11934119 TI - Medically fragile children: an integrative review of the literature and recommendations for future research. AB - The literature related to medically fragile children (MFC) is analyzed, and a model is generated to improve quality of care and cost effectiveness. The sources of stress for families include home care professionals, respite care, financial concerns, and limited community resources. Children cared for in hospitals often experience less than desirable quality outcomes. Community-based pediatric extended care facilities may be a means of reducing family stress, improving physiologic and developmental outcomes, and reducing cost. Case management may significantly reduce parental stress and improve the quality of life for these children. Additional systematic study of care options for MFC is essential. PMID- 11934120 TI - Maternal limit-setting patterns and toddler development of self-concept and social competence. AB - In 126 mother-toddler dyads, the relationships between maternal limit-setting patterns at 12, 24, and 36 months were examined in relation to toddler self concept and social competence at 3 years. Maternal limit-setting patterns differentiated toddler self-concept and social competence scores, demonstrating effects of socialization as well as cognitive maturation. Less optimal toddler outcomes at 36 months were associated with an inconsistent maternal limit-setting style and an indirect style. Children of mothers with a teaching-based pattern yielded the most positive outcomes. Although a power-based maternal limit-setting style was related to diminished self-regulatory behavior in a concurrent limit setting context, outcomes for children of power-based mothers in terms of self concept and social competence were not as negative as expected. PMID- 11934121 TI - Middle-class mothers' perceptions of peer and sibling victimization among children with Asperger's syndrome and nonverbal learning disorders. AB - This article describes the yearly prevalence and frequency of peer and sibling victimization as reported by a large national sample of middle-class mothers of children with Asperger's syndrome and nonverbal learning disorders. An anonymous, mailed survey was sent to families solicited from two national Internet sites for parents of children with Asperger's and nonverbal learning disorders using the Comprehensive Juvenile Victimization scale and three questions designed to measure peer shunning. The overall prevalence rate reported by mothers of peer victimization was 94%. Mothers reported that almost three-quarters of their children had been hit by peers or siblings in the past year and 75% had been emotionally bullied. On the more severe end of peer victimization, 10% of the children were attacked by a gang in the past year and 15% were victims of nonsexual assaults to the genitals. Peer shunning also was common. A third of the children had not been invited to a single birthday party in the past year, and many were eating alone at lunch or were picked last for teams. Peer shunning was significantly correlated with peer bullying and assault. The high rates of peer shunning and peer victimization reported suggest that children with Asperger's and nonverbal learning disorders may require further scrutiny and attention concerning their victimization experiences by peers and siblings. Implications for nursing professionals are reviewed. PMID- 11934122 TI - Perceptions of parents' participation in the care of their child undergoing day surgery: pilot-study. AB - In Canada, more than 100,000 children have procedures in day surgery units. In spite of certain programs designed to prepare those children and their parents for the procedures and events that take place in a hospital, the parents' role during the day stay is not openly and directly approached. Although many pediatric centers recognize the importance of the role of the family in the life of children with health problems, the perceptions of parents with children undergoing day surgery and their role in the hospital have not yet been studied. Therefore, the objective of our study was to describe parents' perceptions regarding their participation in the care of their children in a day surgery. Godin and Kok's (1996) integrated behavior prediction model related to health, which was drawn from Ajzen's theory of planned behavior (1991), was chosen for this research. An interview questionnaire was developed as the measurement instrument. Content analysis was the basis for the analysis of the data gathered through the interview questionnaire. Our article presents the results of this pilot study. PMID- 11934123 TI - Helping children cope with tragedy. PMID- 11934124 TI - Perceptual illusions in eating disorders: rigid and fluctuating styles. AB - This study investigated perceptual styles in anorexia nervosa (AN) and bulimia nervosa (BN) using a perceptual set task. We hypothesised that, consistent with personality style research. AN patients might be more rigid in style than those with BN or no eating disorder. We found that once an illusion had been established, participants with AN and BN showed more illusions than non-ED women. However, while AN patients responded rigidly, giving the same response repeatedly, BN patients were more likely to change their responses. The study suggests interesting differences to be followed up in future research. Differences in rigid and fluctuating perceptual styles may have implications for understanding the phenomenology of eating disorders, and have implications for treatment. PMID- 11934125 TI - Relations between anxiety sensitivity and panic symptoms in nonreferred children and adolescents. AB - Anxiety sensitivity (AS), the fear of anxiety-related sensations, has been posited to be a cognitive risk factor for the development of anxiety disorders but has been understudied in youth. The purpose of the present investigations was to evaluate relations between AS and panic symptoms in nonreferred children and adolescents. In Study 1, (N = 113, mean age, 13.98). scores on the Childhood Anxiety Sensitivity Index (CASI) predicted the experience of uncued panic attacks after controlling for general anxiety and depression, although the total variance accounted for was small. In Study 2 (N = 52; mean age, 9.48), the Panic/ Agoraphobia subscale of the Spence Children's Anxiety Scale was used as the criterion variable. CASI score again predicted panic symptoms after controlling for trait anxiety and depression. Identification of a risk factor for panic attacks and panic disorder in youth will have important implications for etiologic theory, intervention, and prevention. PMID- 11934126 TI - Another White Christmas: fantasy proneness and reports of 'hallucinatory experiences' in undergraduate students. AB - In the current experiment, 44 undergraduate students were asked to listen to white noise and instructed to press a button when they believed hearing a recording of Bing Crosby's White Christmas without this record actually being presented. Fourteen participants (32%) pressed the button at least once. These participants had higher scores on fantasy proneness and the Launay-Slade Hallucination Scale (LSHS) compared to participants without hallucinatory reports. Both groups did not differ in terms of imagery vividness or sensitivity to social demands. Logistic regression suggested that fantasy proneness is a better predictor of hallucinatory reports than are LSHS scores. This might imply that hallucinatory reports obtained during the White Christmas test reflect a non specific preference for odd items rather than schizophrenia-like, internal experiences. PMID- 11934127 TI - Comorbidity fails to account for the relationship of expressed emotion and perceived criticism to treatment outcome in patients with anxiety disorders. AB - This study was designed to test the hypothesis that comorbidity mediates the relationship between expressed emotion, perceived criticism (PC), and treatment outcome in anxiety disorders. Although comorbid major depression and Axis II traits were related to poor outcome in agoraphobic and obsessive-compulsive outpatients, these variables did not prove to be mediators of drop-out or treatment outcome. A secondary aim was to examine whether PC predicted change in symptoms of depression. Even when posttreatment anxiety severity and comorbidity were controlled, PC accounted for a significant proportion of the variance in change on the Symptom Checklist-90-R depression scale. PMID- 11934128 TI - A priori expectancy bias and its relation to shock experience and anxiety: a naturalistic study in patients with an automatic implantable cardioverter defibrillator. AB - Patients with an automatic implantable cardioverter defibrillator (AICD) may offer an unique naturalistic opportunity to study whether expectancy biases develop because of precipitating aversive or traumatic experiences and/or because of elevated anxiety. An expectancy bias and its associations with AICD discharge and anxiety was examined in 24 AICD patients with a thought experiment. While patients without AICD discharge exhibited no expectancy bias, patients with discharge experiences were found to expect that stimuli depicting medical emergency situations will be followed by an aversive consequence. The magnitude of their expectancy bias was positively correlated with their anxiety level. In the group with AICD discharge, patients with low anxiety levels exhibited no bias, while patients with high anxiety levels exhibited a rather strong bias. It seems that the experience of an aversive or traumatic event, here an AICD discharge, is a necessary (but not sufficient) precipitating event for the development of an expectancy bias. If such an event happens, trait anxiety level presumably determines if and how strong the expectancy bias will be. PMID- 11934129 TI - Reactivity to interoceptive cues in nocturnal panic. AB - In this study, patients with panic disorder (PD) who suffered nocturnal panic (NP) attacks were compared with PD patients who never experienced NP attacks and healthy controls. Three tasks were chosen to evaluate attention to cardiac cues, reactivity to induction of respiratory cues, and reactivity to relaxation cues. Relative to healthy controls, PD groups reported more fear of all three tasks and showed more physiological arousal in response to the hyperventilation task. The only task on which the two PD groups differed was the relaxation task, where nocturnal panickers were significantly more distressed. These findings are consistent with the notion that nocturnal panickers are fearful of states involving a diminution of conscious awareness or vigilance. PMID- 11934130 TI - Application of hazard analysis critical control points (HACCP) to organic chemical contaminants in food. AB - Hazard Analysis Critical Control Points (HACCP) is a systematic approach to the identification, assessment, and control of hazards that was developed as an effective alternative to conventional end-point analysis to control food safety. It has been described as the most effective means of controlling foodborne diseases, and its application to the control of microbiological hazards has been accepted internationally. By contrast, relatively little has been reported relating to the potential use of HACCP, or HACCP-like procedures, to control chemical contaminants of food. This article presents an overview of the implementation of HACCP and discusses its application to the control of organic chemical contaminants in the food chain. Although this is likely to result in many of the advantages previously identified for microbiological HACCP, that is, more effective, efficient, and economical hazard management, a number of areas are identified that require further research and development. These include: (1) a need to refine the methods of chemical contaminant identification and risk assessment employed, (2) develop more cost-effective monitoring and control methods for routine chemical contaminant surveillance of food, and (3) improve the effectiveness of process optimization for the control of chemical contaminants in food. PMID- 11934131 TI - Microbial control by packaging: a review. AB - Since early man first used a variety of natural containers to store and eat foods, significant developments in food packaging materials have provided the means to suppress microbial growth as well as protect foods from external microbial contamination. Throughout this progression, packaging materials have been developed specifically to prevent the deterioration of foods resulting from exposure to air, moisture, or pH changes associated with the food or the surrounding atmosphere. Both flexible and rigid packaging materials, alone or in combination with other preservation methods, have been developed to offer the necessary barrier, inactivation, and containment properties required for successful food packaging. Examples of flexible packaging used to inactivate microorganisms associated with foods include controlled atmosphere, vacuum, modified atmosphere, active, and edible packaging. Additionally, the combination of rigid packaging materials made from metal, glass, or plastic with heat provides the most effective and widely used method for inactivating microorganisms. As with all food products, it is necessary to integrate a HACCP based program to assure quality throughout the packaging operation. In addition to packaging improvements, other novel technologies include the development of detectors for oxygen levels, bacterial toxins, and microbial growth, or the integration of time-temperature indicators for detection of improper handling or storage. PMID- 11934132 TI - A functional food product for the management of weight. AB - More than half of Americans have a body mass index of 25 kg/m2 or more, which classifies them as overweight or obese. Overweight or obesity is strongly associated with comorbidities such as type 2 diabetes mellitus, hypertension, heart disease, gall bladder disease, and sleep apnea. Clearly, this is a national health concern, and although about 30 to 40% of the obese claim that they are trying to lose weight or maintain weight after weight loss, current therapies appear to have little effect. None of the current popular diets are working, and there is room for innovation. With the advancing science of nutrition, several nutrients - low-glycemic-index carbohydrates, 5-hydroxytryptophan, green tea extract, and chromium - have been identified that may promote weight loss. The first two nutrients decrease appetite, green tea increases the 24-h energy expenditure, and chromium promotes the composition of the weight lost to be fat rather than lean tissue. These have been assembled in efficacious doses into a new functional food product and described in this review. The product is undergoing clinical testing; each component has already been shown to promote weight loss in clinical trials. PMID- 11934133 TI - Pediocin PA-1, a wide-spectrum bacteriocin from lactic acid bacteria. AB - Pediocin PA-1 is a broad-spectrum lactic acid bacteria bacteriocin that shows a particularly strong activity against Listeria monocytogenes, a foodborne pathogen of special concern among the food industries. This antimicrobial peptide is the most extensively studied class Ila (or pediocin family) bacteriocin, and it has been sufficiently well characterized to be used as a food biopreservative. This review focuses on the progress that have been made in the elucidation of its structure, mode of action, and biosynthesis, and includes an overview of its applications in food systems. The aspects that need further research are also addressed. In the future, protein engineering, genetic engineering and/or chemical synthesis may lead to the development of new antimicrobial peptides with improved properties, based on some features of the pediocin PA-1 molecule. PMID- 11934134 TI - Tuberculosis in households of index patients: is there another way to control tuberculosis? PMID- 11934135 TI - Smoking in Syria: profile of a developing Arab country. AB - One of the main obstacles to tobacco control in the Middle East lies in the shortage of reliable, standardised data on the spread and patterns of tobacco use in society. In Syria, a project aiming at drawing an epidemiological map of the tobacco epidemic in this country was started 4 years ago. Overall, nine studies have resulted, with a total of 6780 participants. The crude prevalence of current smoking among adults in Syria, based on combined information from all studies, is 48% and 9% for males and females, respectively. The prevalence of current smoking among high school adolescents is 16% and 7% for boys and girls, respectively, and was strongly associated with parental and sibling smoking. High school students from families with parents and/or siblings who smoked were 4.4 times more likely to be current smokers than those from non-smoking families. The biggest influx of new smokers among males in Syria is occurring in the early twenties, but an earlier pattern can occur among youths with low academic performance or socioeconomic status. Smoking in women, evaluated by data from physicians, tends to start later than in men and continues to increase with age. Women's smoking in Syria is related to their level of social liberalisation. Data show that active smoking is associated with an increased risk of respiratory diseases among smokers, and that exposure to environmental tobacco smoke (ETS) is associated with an increased risk of respiratory symptoms in children. Knowledge about the harmful effects of smoking and the desire to quit are disproportionate to the rate of successful cessation. The evidence collected indicates possible avenues for tobacco control in Syria, including price increases, smoking cessation programmes, restriction of adolescents' access to cigarettes, and intensive prevention work among women. PMID- 11934136 TI - Implementation of the DOTS strategy for tuberculosis in the Leningrad Region, Russian Federation (1998-1999). AB - SETTING: The DOTS pilot project for tuberculosis control in the Leningrad Region of the Russian Federation, supported by the Finnish Lung Health Association and the World Health Organization (WHO). OBJECTIVE: To assess the efficacy of WHO recommended standard short-course chemotherapy in newly detected pulmonary tuberculosis cases positive by smear or with extensive lung lesions suggestive of culture positivity, under project conditions. METHODS: Analysis of data on case detection, sputum smear conversion and treatment outcome based on standardised (WHO) registers from districts and a central computerised database. RESULTS: Of 859 adult pulmonary tuberculosis cases (292 smear-positive) notified in the Leningrad Region in the study period, 312 new cases were included in the project. The sputum conversion rate at the end of the second month was 82.8% and 91.1% at the end of the third month. Of bacteriologically confirmed cases, 71.3% were successfully treated, 4.9% died, 11.7% defaulted and 8.1% failed. CONCLUSION: In the first year of the pilot project in the Leningrad Region, the DOTS strategy revealed feasibility and moderate efficacy among new pulmonary tuberculosis cases who were either smear-positive or showed extensive lung lesions on chest X-ray, and who were therefore of high epidemiological and medical priority. PMID- 11934137 TI - Epidemiological transition of tuberculosis and future agenda of control in Japan: results of the Ad-Hoc National Survey of Tuberculosis 2000. AB - SETTING: In Japan, the incidence of tuberculosis (TB), which had improved continuously from 1951, has shown an upturn since 1997. OBJECTIVE: An Ad-Hoc National Survey was conducted to review the issues of TB control in Japan and to discuss the future direction of change. METHODS: TB registries kept at health centres in all prefectures were reviewed for the year 1998. Vital statistics and BCG vaccination records were reviewed separately. RESULTS: The survey revealed a 34.7% under-estimation of TB deaths in TB registers, and a 11.2% over-estimation of TB cases registered due to a change in diagnosis after registration. For treatment, the four-drug regimen advocated for smear-positive cases was used in only 54.8% of indicated cases, and treatment duration was very long; for example, 18.4% of first-time hospitalised cases over 15 years of age received in-patient care for more than 6 months and 29.8% continued treatment for more than 12 months. Apart from an increasingly elderly population, other factors contributing to an increase in TB included underlying health risk factors for increased susceptibility to TB and socio-economic risk factors for treatment failure. Regarding preventive measures, active case finding by health screening contributed to detection of only 8.7% of new cases aged over 60 years, but BCG vaccination appeared to be preventive among infants under 1 year of age. CONCLUSION: The National Survey revealed a changing profile of TB in Japan and suggested a need to review and revitalise the TB control programme. PMID- 11934138 TI - Tuberculosis in Thai prisons: magnitude, transmission and drug susceptibility. AB - BACKGROUND: Because of the human immunodeficiency virus (HIV) epidemic, tuberculosis has reemerged as a major public health problem in Thailand. Prison inmates are at high risk for developing tuberculosis because of the high prevalence of HIV infection. OBJECTIVES: To determine the magnitude, transmission, and drug susceptibility of tuberculosis in Thai prisons. SETTINGS: Four provincial prisons in Southern Thailand. DESIGN: Cross-sectional, descriptive, clinical and molecular study. RESULTS: Miniature chest roentgenograms were performed on 304 (6.4%) of 4751 inmates screened for a > or = 2 week history of chronic cough and fever. At least 17 (35%) of 49 inmates who had a miniature chest roentgenogram compatible with tuberculosis were HIV positive. The prevalence of smear-positive pulmonary tuberculosis was 568 per 100,000 inmates, which was eight times higher than that in the general population. Eight (38%) of 21 culture-positive Mycobacterium tuberculosis isolates had DNA fingerprints matching those of another inmate who was housed in the same room or in the same dormitory unit; 39% of the M. tuberculosis isolates were resistant to isoniazid; three of these isolates were also borderline resistant to rifampicin. CONCLUSION: The prevalence of pulmonary tuberculosis in these prisons was high. A substantial proportion were acquired in the prisons. Isoniazid (INH) resistance was common, and theoretically precludes the use of INH preventive therapy for contacts of these cases. Active case finding should be done and directly observed therapy implemented to prevent the spread of tuberculosis into the community. PMID- 11934139 TI - Dosages of anti-tuberculosis medications in the national tuberculosis programs of Kenya, Nepal, and Senegal. AB - SETTING: National tuberculosis programs in Kenya, Nepal, and Senegal. OBJECTIVES: To ascertain adequacy of initial prescriptions of dosages of anti-tuberculosis medications in the three national tuberculosis programs. METHODS: Collection of patient treatment cards in a representative sample of treatment centers in Kenya, Nepal, and Senegal. Calculation of drug dosages in milligram per kilogram body weight of isoniazid, rifampicin, and pyrazinamide and comparison with international recommendations for dosage of these medications. RESULTS: A total of 12,346 patient treatment cards were available. Yet of these only 8640 were analyzed: 5575 (65% of total) from Kenya, 612 (53% of total) from Nepal, and 2453 (95% of total) from Senegal had the patient's weight recorded and were given a nationally recommended treatment regimen. The proportions of patients receiving an internationally recommended isoniazid dosage were 34%, 15%, and 15%, respectively in Kenya, Nepal and Senegal; the corresponding figures for rifampicin were 77%, 77%, and 93% and for pyrazinamide 25%, 3% and 75%, respectively, in the three countries. The majority of errors were over-dosage, but some cases of under-dosage were also identified. CONCLUSIONS: This study shows that over-dosage was a frequent event in all three countries. Two major reasons for this error are inadequate drug combinations in Kenya and Senegal, and in all three countries recommendations for weight brackets that did not ideally fit internationally recommended dosages. It is vital to address these problems to reduce both the risk of unnecessary drug toxicity on one end of the spectrum, and suboptimal drug levels on the other. PMID- 11934140 TI - Optimal tuberculosis case detection by direct sputum smear microscopy: how much better is more? AB - SETTING: A tuberculosis control project in Bangladesh. OBJECTIVE: To define the efficiency of numbers of microscopic fields screened and the sputum collection scheme used for diagnostic smear examination. DESIGN: Quality controllers noted cumulative numbers of acid-fast bacilli per 100 fields screened. The incremental diagnostic yield of different sputum sampling strategies was determined. Doubtful series were re-checked and/or further samples examined. RESULTS: Acid-fast bacilli were found in 99.6% of 1412 positive and in 79.3% of 576 scanty slides in the first 100 fields. Examination of a third specimen yielded a maximum of 2.7% positives incrementally. The most efficient strategy, using three morning specimens, yielded 94.2% positives on the first and 1.0% on the third sputum; although 10% of suspects did not return, only 1.5% of the positives were among them and more cases were confirmed and treated. The positive predictive value of a single positive or scanty smear was very high (99.2%). CONCLUSIONS: Reading more than 100 fields per smear or examining a third sputum has insufficient marginal returns to justify the workload. Examining morning samples only is more efficient, and their collection does not necessarily inconvenience patients. Treatment can be started on the basis of one positive smear. Provided that a well functioning system of smear-microscopy quality control is in place, we propose a strategy based on examination of two morning sputum samples for negative suspects, with the diagnosis based on a single positive result. PMID- 11934141 TI - Tuberculosis should not be considered an AIDS-defining illness in areas with a high tuberculosis prevalence. AB - OBJECTIVE: To assess the prognosis of human immunodeficiency virus (HIV) associated tuberculosis in a high tuberculosis prevalence setting. METHODS: Survival and subsequent AIDS-defining illnesses (ADI) of patients with tuberculosis were compared with patients remaining tuberculosis-free in a prospective cohort study in two university-affiliated adult HIV clinics in Cape Town. RESULTS: Tuberculosis without prior or simultaneous ADI was diagnosed in 141 patients. CD4+ T-lymphocyte count was >200 cells/microl in 67% of the incident tuberculosis cases. Survival in tuberculosis patients was comparable to that of patients with oral hairy leukoplakia and/or oral candidiasis (median = 23.6 vs. 27.8 months respectively; P = 0.59, adjusted hazard ratio [AHR] = 0.87; 95%CI 0.63-1.58), and better than in patients with AIDS (median = 11.5 months; P < 0.001, AHR = 2.37; 95% CI 1.93-4.66). Subsequent ADI were less frequent in tuberculosis than in AIDS patients (AHR = 0.36; 95%CI 0.23-0.58). Survival of patients with pulmonary or extra-pulmonary tuberculosis was similar (P = 0.32). CONCLUSION: Tuberculosis in HIV-infected patients from areas endemic with tuberculosis occurs across a wide spectrum of immune suppression and has a considerably better prognosis than other ADI. Inclusion of tuberculosis in the clinical case definition of AIDS in such areas should be reconsidered. PMID- 11934142 TI - Yield of continued monthly sputum evaluation among tuberculosis patients after culture conversion. AB - SETTING: New York City. OBJECTIVE: To evaluate the yield of continued monthly sputum monitoring after culture conversion. DESIGN: A retrospective review of tuberculosis patients verified between 1 January 1995 and 31 December 1996 who had: 1) pulmonary tuberculosis with organisms susceptible to isoniazid and rifampin; 2) culture conversion; and 3) completed therapy. We assessed time to smear and culture conversion and number of persons who developed a positive culture after culture conversion (culture reversion). RESULTS: Of 1440 patients, 379 were cared for by tuberculosis control program providers and 1061 were cared for by other providers; 813 (56%) were initially smear-positive. After the fifth month, 44 (5.3%) were smear-positive; four of these were culture-positive. Eighteen (1.3%) had culture reversions; eight were smear-positive. Excluding one specimen per patient collected at treatment completion, 7967 sputum samples were collected after culture conversion. The minimum estimated cost per culture reversion detected was $26,557. CONCLUSION: Continued monthly monitoring of sputum after culture conversion identified a very small number of patients who had culture reversion. However, patients who cannot tolerate or adhere to a standard regimen may need continued monitoring to assess response to treatment. For all patients a specimen should be collected at the end of treatment to document cure. PMID- 11934143 TI - Evaluation of a rapid-format antibody test and the tuberculin skin test for diagnosis of tuberculosis in two contrasting endemic settings. AB - OBJECTIVE AND SETTING: We evaluated a rapid-format antibody card test and the tuberculin skin test for diagnosis of active tuberculosis (TB) in high (Cairo, Egypt) and low (St. Louis, USA) prevalence areas. DESIGN: Prospective study of hospitalized TB patients and controls with other chest diseases. RESULTS: Test performance varied significantly in the two study sites. The antibody test detected 87% of 71 smear-positive pulmonary TB cases (86% of smear-negative pulmonary cases and 48% of TB meningitis cases) in Egypt; specificity was 82%. The tuberculin test was highly sensitive in Egypt in subjects with pulmonary TB (100%) but not in those with meningitis (23%); specificity was 70%. The sensitivity and specificity of the antibody test in St. Louis were 29% and 79%, respectively; 50% of St. Louis TB cases and 15% of controls had positive tuberculin tests. CONCLUSIONS: This convenient antibody card test may have value for diagnosis of patients suspected of having TB in high prevalence areas like Egypt. However, the specificity of the test is too low for it to be useful as a screening test. Our results suggest that neither the antibody test nor the tuberculin test have much diagnostic utility in low prevalence settings like St. Louis. PMID- 11934144 TI - Chemotherapeutic treatment for spinal tuberculosis. AB - AIM: To evaluate whether 6 months of chemotherapy for patients with spinal tuberculosis prevents relapse as effectively as more than 6 months of chemotherapy. METHOD: Literature review. Medline search including references, from January 1978 to November 2000. Inclusion criteria for publications: diagnosis of spinal tuberculosis confirmed bacteriologically and/or histologically, or probable on the basis of clinical and radiological parameters; treatment regimen (whether or not in combination with surgery) included isoniazid (H), rifampicin (R) and pyrazinamide (Z); follow-up period after completion of treatment of 12 months or more. EXCLUSION CRITERIA: patients with relapse who had previously been treated adequately for tuberculosis. OUTCOME PARAMETERS: Relapse rate. RESULTS: Four publications were found with HRZ regimens of 6 months' duration and 10 publications with HRZ regimens of >6 months' duration. A number of patients had received HRE (E = ethambutol) for > or = 9 months. In the results, no distinction was made between treatment groups. HRZ for 6 months led to a relapse rate of 0% (0/56, 95%CI 0.0-6.4); follow-up after surgical intervention ranged from 6 to 108 months. HRZ for > or = 9 months (> or = 119 patients) or HRE for > or = 9 months (< or = 71 patients) led to a relapse rate of 2% (4/218, 95%CI 0.6-5.0); follow-up after surgical intervention was 6-168 months. Despite the small number of studies, 6 months of therapy is probably sufficient for patients with spinal tuberculosis. PMID- 11934145 TI - High frequency of tuberculosis in households of index TB patients. AB - A case-control study was carried out in 44 non-private hospitals in Malawi which register and treat patients with tuberculosis (TB), to determine whether households of index TB patients had an increased frequency of TB compared with households where no TB had been diagnosed. Interviews were conducted in 770 smear positive PTB patients and 918 control patients from antenatal, orthopaedic or surgical wards. TB patients and controls came from households which were similar in terms of type of house, household cash income and family size. In the previous 12 months, 52 (7%) TB patients had household members who developed TB compared with 11 (1%) control patients (OR 5.97, 95%CI 2.99-12.21). Of 2766 household members of TB patients, 56 had developed TB in the previous 12 months (frequency = 2024/100,000), which was significantly higher than the 11/4121 household members of control patients (frequency = 343/100,000, P < 0.001). There is a higher frequency of TB in households of index TB patients. PMID- 11934146 TI - Washing of new microscopic glass slides in dichromate solution does not influence sputum AFB smear results. AB - SETTING: Microscopy centres in Tiruvallur District, Tamil Nadu, India, implementing DOTS. OBJECTIVE: To know whether washing new glass slides in dichromate solution is essential for effective sputum acid-fast bacilli microscopy. METHODS: Two direct smears were prepared from each of 1750 sputum samples. One was made on dichromate solution-cleaned new glass slides and the other was made on unwashed new glass slides. The smears were blinded and examined. RESULTS: Of the 1750 specimens, 13.5% and 13.08% were positive for AFB using washed and unwashed slides, respectively (P = 0.12). The concordance between these two (including one grade above and one below) was 98.7%. CONCLUSION: Washing of new glass slides in dichromate solution is not essential for AFB microscopy. PMID- 11934147 TI - Chest radiography in the diagnosis of pneumoconiosis. PMID- 11934148 TI - Pancreatic lineage analysis using a retroviral vector in embryonic mice demonstrates a common progenitor for endocrine and exocrine cells. AB - The origin of pancreatic endocrine cells is unknown. Some studies have suggested that there is a common pancreatic progenitor which gives rise to both endocrine and exocrine cells, while others have suggested separate endocrine and exocrine lineages. Previous conclusions have been based on indirect data, such as the co expression of molecular markers. We directly assessed the relationship between endocrine and exocrine cells during development using a lineage tracer. A replication-incompetent retrovirus was used to introduce the reporter gene alkaline phosphatase into single cells in explants of mouse embryonic pancreas. After a week in culture, the subsequent fate of the infected cells could then be determined. The results show that a common pancreatic progenitor cell exists, which gives rise to both endocrine and exocrine cells. PMID- 11934149 TI - Colinearity and non-colinearity in the expression of Hox genes in developing chick skin. AB - Hox genes are usually expressed temporally and spatially in a colinear manner with respect to their positions in the Hox complex. We found that these characteristics apply to several Hox genes expressed in developing chick skin (Hoxb-4, Hoxa-7 and Hoxc-8), and we classed this group of genes as regionally restricted. To our surprise, we found that most of the Hox genes we examined are regionally unrestricted in their expression in the embryonic chick skin. This second group includes the Hoxd genes, Hoxd-4 to Hoxd-13, Hoxa-11 and Hoxc-6. Temporally, the expression of the regionally restricted genes can be observed by E5 within the epidermis, whereas the spatially unrestricted genes are not expressed in the epidermis until E6.25. Unexpectedly, we found that all the unrestricted genes are expressed concomitantly and therefore do not conform to temporal colinearity. Moreover, the dermal expression for both groups occurs later, but maintains the same anteroposterior patterning to that seen previously in the epidermis. During embryonic day 7-8, expression for all genes is up regulated within the dense dermis whilst being reduced within the inter-bud regions. Later expression within the bud mesenchyme is down-regulated whilst high levels of transcriptional activity are detectable within the epidermal sheath of each feather bud. These results indicate that the transcriptional activity of Hox genes in the developing chick skin could be important during embryonic skin patterning both by providing regionally restricted positional cues, and also by imparting generic signals necessary for feather morphology. PMID- 11934150 TI - Multiple interactions between maternally-activated signalling pathways control Xenopus nodal-related genes. AB - We have investigated the induction of the six Xenopus nodal-related genes, Xnr1 Xnr6, by maternal determinants. The beta-catenin pathway was modelled by stimulation using Xwnt8, activin-like signalling was modelled by activin, and VegT action was studied by overexpression in animal cap explants. Combinations of factors were examined, and previously unrecognised interactions were revealed in animal caps and whole embryos. For the induction of Xnr5 and Xnr6 in whole embryos, using a beta-catenin antisense morpholino oligonucleotide or a dominant negative XTcf3, we have demonstrated an absolute permissive requirement for the beta-catenin/Tcf pathway, in addition to the requirement for VegT action. In animal caps Xnr5 and Xnr6 are induced in response to VegT overexpression, and this induction is dependent upon the concomitant activation of the beta-catenin pathway that VegT initiates in animal caps. For the induction of Xnr3, VegT interacts negatively so as to inhibit the induction otherwise observed with wnt signalling alone. The negative effect of VegT is not the result of a general inhibition of wnt-signalling, and does not result from an inhibition of wnt induced siamois expression. A 294 bp proximal promoter fragment of the Xnr3 gene is sufficient to mediate the negative effect of VegT. Further experiments, employing cycloheximide to examine the dependence of Xnr gene expression upon proteins translated after the mid-blastula stage, demonstrated that Xnrs 4, 5 and 6 are 'primary' Xnr genes whose expression in the late blastula is solely dependent upon factors present before the mid-blastula stage. PMID- 11934151 TI - Lamina-associated polypeptide 2 (LAP2) expression in fish and amphibians. AB - Somatic and germinal cells of 15 fish and 33 amphibian species were examined by SDS-PAGE followed by immunoblotting to determine the expression of LAP2 (lamina associated polypeptide 2). LAP2 expression in frogs, salamanders and fish does not vary with the mode of reproduction. In fish and frog cells, a rim-like LAP2 positive region was detected around the nucleus by indirect immunofluorescence microscopy. The cell distribution and expression patterns of LAP2 in fish, frogs and salamanders are comparable with those found in Xenopus and zebrafish. The mammalian somatic cell pattern, which may also occur in gymnophione amphibians, includes LAP2alpha, beta and gamma as major isoforms, whereas LAP2alpha does not occur in cells of fish, frogs and salamanders. In fish, LAP2gamma is the major isoform of somatic cells, suggesting that LAP2gamma may be ancestral. However, in the rainbow trout, as in frogs and salamanders, LAP2beta was the major somatic isoform. Fish and frog sperm only express low molecular weight polypeptides. In contrast, fish and frog oocytes express an oocyte-specific LAP2 isoform of high molecular weight. In the toad Bufo marinus this isoform becomes upregulated in pre-vitellogenic oocytes of 150-200 microm in diameter. The absence of LAP2alpha and the differential expression of LAP2 isoforms in somatic and germ cells, as found in fish and frogs, may be ancestral vertebrate characters. In spite of differences in developmental time, the LAP2 isoforms of somatic cells are upregulated during gastrulation, suggesting that LAP2 may be implicated in the early development of fish and frog. PMID- 11934152 TI - Involvement of myogenic regulator factors during fusion in the cell line C2C12. AB - The myogenic factors, MyoD, myogenin, Myf5 and MRF4, can activate skeletal muscle differentiation when overexpressed in non-muscular cells. Gene targeting experiments have provided much insight into the in vivo functions of MRF and have defined two functional groups of MRFs. MyoD and Myf5 may be necessary for myoblast determination while myogenin and MRF4 may be required later during differentiation. However, the specific role of these myogenic factors has not been clearly defined during one important stage of myogenesis: the fusion of myoblasts. Using cultured C2C12 mouse muscular cells, the time-course of these proteins was analyzed and a distinct expression pattern in fusing cells was revealed. In an attempt to clarify the role of each of these regulators during myoblast fusion, an antisense strategy using oligonucleotides with phosphorothioate backbone modification was adoped. The results showed that the inhibition of myogenin and Myf5 activity is capable of significantly preventing fusion. Furthermore, the inhibition of MyoD can wholly arrest the engaged fusion process in spite of high endogenous expression of both myogenin and Myf5. Consequently, each MRF seems to have, at this defined step of myogenesis, a specific set of functions that can not be substituted for by the others and therefore may regulate a distinct subset of muscle-specific genes at the onset of fusion. PMID- 11934154 TI - Notch is required for outgrowth of the Xenopus tail bud. AB - It has previously been shown that Notch, Delta and Lunatic Fringe are expressed together in the leading edge of the tail bud of the Xenopus embryo prior to outgrowth (Beck and Slack, 1998). It has also been shown that ectopic expression of a constitutive form of Notch, Notch-ICD, will provoke ectopic tail formation (Beck and Slack, 1999). Here we show that inhibition of Notch activity in vivo prevents outgrowth of the tail bud. This is achieved using inhibitors of the protease that carries out the ligand-induced intramembranous cleavage of Notch. Other protease inhibitors that do not inhibit Notch cleavage do not affect tail outgrowth. PMID- 11934153 TI - Signalling via type IA and type IB bone morphogenetic protein receptors (BMPR) regulates intramembranous bone formation, chondrogenesis and feather formation in the chicken embryo. AB - Bone morphogenetic proteins (BMPs) signal via complexes of type I and type II receptors. In this study, we mapped the expression of type IA, type IB and type II receptors during craniofacial chondrogenesis and then perturbed receptor function in vivo with retroviruses expressing dominant-negative or constitutively active type I receptors. BmprIB was the only receptor expressed within all cartilages. BmprIA was initially expressed in cartilage condensations, but later decreased within cartilage elements. BmprII was expressed at low levels in the nasal septum and prenasal cartilage and at higher levels in other craniofacial cartilages. The maxillary prominence, which gives rise to several intramembranous bones, expressed both type I receptors. Misexpression of dnBMPRIB decreased the size of cartilages and bones on the treated side. In contrast, dnBMPRIA had no effect on the skeletal phenotype. The phenotypes of caBMPRIA and caBMPRIB were similar; both led to overgrowth of cartilage elements, thinner bones with fewer trabeculae and inhibition of feather development. Infection with constitutively active viruses resulted in ectopic expression of Msx1, Msx2 and Fgfr2 throughout the maxillary mesenchyme. These data suggest that the pattern of trabeculation in membranous bones derived from the maxillary prominence was related to the change in expression pattern and that Msx and Fgfr2 genes were downstream of both type I BMP receptors. We conclude that the requirement for the type IB is greater than for the type IA receptor but, when active, both receptors play similar roles in regulating bone, cartilage and feather formation in the skull. PMID- 11934155 TI - Goosecoid and cerberus-like do not interact during mouse embryogenesis. AB - Mouse Cerberus-like (Cer-l) is a neural inducer molecule, capable of inhibiting Nodal and BMP-4 signals in the extracelular space. The cer-l expression domain in the Anterior Visceral Endoderm (AVE) and prechordal plate, tissues involved in head induction and patterning, respectively, suggested a role for this gene in head formation. However, animals homozygous for the cer-l null allele failed to show any abnormality, leading us to propose the existence of other factor(s) that might compensate for cer-l loss-of-function. Since goosecoid (gsc) shares some domains of expression with cer-l and was shown to be essential for head morphogenesis, we tested its ability to interact genetically with cer-l. With this aim we generated cer-l;gsc double mutants. These animals were analyzed at birth for skeletal defects and revealed the same phenotype as gsc-/- single mutants. We also investigated the proper patterning of structures adjacent to the prechordal plate by performing in situ hybridization of HNF-3beta, Six-3 and BF 1, genes whose expression domains remained unchanged. In conclusion, the analysis carried out indicated that gsc does not compensate for cer-l loss-of-function and that these genes do not interact genetically. PMID- 11934156 TI - Melanoma Cell Adhesion Molecule (MCAM) expression in the myogenic lineage during early chick embryonic development. AB - We describe the expression pattern of cMCAM, a cell adhesion molecule of the immunoglobulin superfamily, in early chick embryonic development by in situ hybridisation. An initial ectodermal domain of expression is subsequently expanded, and cMCAM is expressed in the neural crest cells, otic vesicle, heart primordium, notochord and endoderm. In addition, cMCAM expression localises in the myotome once the somite cells have been specified. An in vitro murine cellular system allowed us to confirm that MCAM expression coincides with the onset of myogenic cell determination. PMID- 11934157 TI - Using reflectance models for color scanner calibration. AB - We examine the use of linear spectral reflectance models for calibrating a color scanner to generate device-independent CIE XYZ values from scanner vectors. Polynomial regression approaches to color scanner calibration use parameterized functions to approximate the calibration mapping over a set of training colors. These approaches can perform poorly if the parameterized functions do not accurately model the structure of the desired calibration mapping. Several studies have shown that linear reflectance models accurately characterize a wide range of materials. By viewing color scanner calibration as reflectance estimation, we can incorporate linear reflectance models into the calibration process. We show that in most cases linear models do not constrain the calibration problem sufficiently to allow exact recovery of X, Y, Z from a scanner vector obtained with three filters. By examining a series of methods that exploit information about reflectance functions, however, we show that reflectance information can be used to improve the accuracy of calibration over that of standard methods applied to the same set of inputs. PMID- 11934159 TI - Comparison of the spatial-frequency selectivity of local and global motion detectors. AB - Convergent physiological and behavioral evidence indicates that the initial receptive fields responsible for motion detection are spatially localized. Consequently, the perception of global patterns of movement (such as expansion) requires that the output of these local mechanisms be integrated across visual space. We have differentiated local and global motion processes, with mixtures of coherent and incoherent moving patterns composed of bandpass filtered dots, and have measured their spatial-frequency selectivity. We report that local motion detectors show narrow-band spatial-frequency tuning (i.e., they respond only to a narrow range of spatial frequencies) but that global motion detectors show broadband spatial-frequency tuning (i.e., they integrate across a broad range of spatial frequencies), with a preference for low spatial frequencies. PMID- 11934158 TI - Videokeratoscope-line-of-sight misalignment and its effect on measurements of corneal and internal ocular aberrations. AB - A growing number of research laboratories are using the new technologies of videokeratoscopy and Shack-Hartmann aberrometry, in combination, to study the optical structure of the human eye. A potential source of error arises, however, because the two instruments are designed to measure the human eye along different reference axes. The Shack-Hartmann aberrometer is usually aligned coaxially with the line of sight, but videokeratoscopes usually are not. Thus far, corneal optics research has not adequately addressed the problem of videokeratoscope-line of-sight misalignment and its effect on the computation of corneal and internal ocular aberrations. We measured corneal, ocular, and internal aberrations for three normal human eyes, developed a method to compensate for videokeratoscope line-of-sight misalignment, and analyzed the importance of compensating for the misalignment. Our results show that when the value of angle lambda (the angle between the line of sight and the pupillary axis) is larger than 2 degrees-3 degrees, the misalignment, if ignored, can lead to incorrect estimates of corneal and internal aberrations as well as the corneal/internal aberration balance. PMID- 11934160 TI - Role of synchrony in contour binding: some transient doubts sustained. AB - The temporal correlation hypothesis proposes that neurons signal mutual inclusion in complex features, such as extended contours, by phase-locking their firing [C. M. Gray and W. Singer, Proc. Nat]. Acad. Sci. USA 86, 1698 (1989)]. Although this hypothesis remains controversial, a number of recent psychophysical studies have suggested that temporal correlation among features can indeed promote perceptual grouping. In particular, subjects are better at detecting extended visual contours embedded within a field of distractor elements when a small delay is present between a cycling presentation of the contour and the background [Nature 394, 179 (1988)]. We have replicated this finding and examined three potentially confounding factors. First, we controlled local density and used more curved contours composed of bandpass elements to confirm that the effect was associated with contour integration and not with the operation of coarse-scale spatial filters. Second, we minimized the effects of saccadic eye movements (which could combine with the flicker of the asynchronous display to introduce motion cues at the contour location) both by using a fixation marker that was visible only when observers made a saccade (allowing them to reject these trials) and by retinally stabilizing the stimulus. We report that eye movements contribute to the effect. Third, we asked if either visible persistence or transients at the onset and the offset of the asynchronous stimuli might contribute to the effect. We report that the effect is largely abolished by the inclusion of prestimulus and poststimulus masks and is entirely abolished by ramping the contrast of the stimulus on and off. Neither ramping, masking, nor stabilization should specifically disrupt a contour-binding scheme based on temporal synchrony, and we conclude that it is the transient component at the onset and the offset of these stimuli that is responsible for the reported advantage for asynchronous presentation. PMID- 11934161 TI - Determining surface orientations of transparent objects based on polarization degrees in visible and infrared wavelengths. AB - Techniques for modeling an object through observation are very important in object recognition and virtual reality. A wide variety of techniques have been developed for modeling objects with opaque surfaces, whereas less attention has been paid to objects with transparent surfaces. A transparent surface has only surface reflection; it has little body reflection. We present a new method for obtaining surface orientations of transparent surfaces through analysis of the degree of polarization in surface reflection and emission in visible and far infrared wavelengths, respectively. This parameter, the polarization degree of reflected light at the visible wavelengths, is used for determining the surface orientation at a surface point. The polarization degree at visible wavelengths provides two possible solutions, and the proposed method uses the polarization degree at far-infrared wavelengths to resolve this ambiguity. PMID- 11934162 TI - Numerical analysis of plasmon-resonance absorption in bisinusoidal metal gratings. AB - We numerically investigate plasmon-resonance absorption of incident light energy by a bisinusoidal metal grating, i.e., one whose surface profile is sinusoidally corrugated in two orthogonal directions with a common period. Employing Yasuura's modal expansion method, we solve the problem of plane-wave diffraction by the grating and evaluate the absorption, which is observed as dips in diffraction efficiency curves. We examine the field distribution and energy flow in detail at the angles of incidence at which the absorption occurs. We show that the absorption is caused by coupling of the TM component of an evanescent order with surface plasmons. A phase-matching condition is used in the prediction of the incident angle at which the absorption occurs. This, together with the field profile in the presence of the resonance absorption, explains the mechanism of the absorption. We then illustrate interesting features of the absorption: enhancement of polarization conversion between the incident light and the reflected light and simultaneous excitation of two plasmon waves in directions that are symmetric with respect to the plane of incidence. PMID- 11934163 TI - Two-dimensionally-periodic diffractive optical elements: limitations of scalar analysis. AB - The range of validity of the scalar diffraction analysis is quantified for the case of two-dimensionally-periodic diffractive optical elements (crossed gratings). Three canonical classes of two-dimensionally-periodic grating structures are analyzed by using the rigorous coupled-wave analysis as well as the scalar diffraction analysis. In all cases the scalar-analysis diffraction efficiencies are compared with the exact diffraction efficiencies. The error in using the scalar analysis is then determined as a function of the grating period(s)-to-wavelength ratio(s), the minimum feature size, the grating depth, the refractive index of the grating, the incident polarization, and the number of phase levels. The three classes of two-dimensional (2-D) unit cells are as follows: (1) a rectangular pillar, (2) an elliptical pillar, and (3) an arbitrarily pixellated multilevel 2-D unit cell that is representative of more complicated diffractive optical elements such as computer-generated holograms. In all cases a normally incident electromagnetic plane wave is considered. It is shown that the error of the scalar diffraction analysis in the case of two dimensionally-periodic diffractive optical elements is greater than that for the corresponding one-dimensionally-periodic counterparts. In addition, the accuracy of the scalar diffraction analysis degrades with increasing refractive index, grating thickness, and asymmetry of the 2-D unit cell and with decreasing grating period-to-wavelength ratio and feature size. PMID- 11934164 TI - Plane-wave-time-domain-enhanced marching-on-in-time scheme for analyzing scattering from homogeneous dielectric structures. AB - A novel and fast integral-equation-based scheme is presented for analyzing transient electromagnetic scattering from homogeneous, isotropic, and nondispersive bodies. The computational complexity of classical marching-on-in time (MOT) methods for solving time-domain integral equations governing electromagnetic scattering phenomena involving homogeneous penetrable bodies scales as O(NtNs2). Here, Nt represents the number of time steps in the analysis, and Ns denotes the number of spatial degrees of freedom of the discretized electric and magnetic currents on the body's surface. In contrast, the computational complexity of the proposed plane-wave-time-domain-enhanced MOT solver scales as O(NtNs log2Ns). Numerical results that demonstrate the accuracy and the efficacy of the scheme are presented. PMID- 11934165 TI - Scattering from rough inhomogeneous media: splitting of surface and volume scattering. AB - The intensity scattered by particles randomly placed beneath a rough interface is studied with rigorous simulations. It is shown that the angular intensity pattern is close to that obtained by adding the intensity scattered by particles under a flat surface to that scattered by a rough homogeneous surface whose permittivity is evaluated with an effective-medium theory. This heuristic splitting rule is accurate for a large range of parameters that are well beyond any perturbative treatment. PMID- 11934166 TI - Determination of particle size by using the angular distribution of backscattered light as measured with low-coherence interferometry. AB - We employ a novel interferometer to measure the angular distribution of light backscattered by a turbid medium. Through comparison of the measured data with the predictions of Mie theory, we are able to determine the size of the scatterers comprising the medium with subwavelength precision. As the technique is based on low-coherence interferometry, we are able to examine the evolution of the angular distribution of scattered light as it propagates into the medium. The effects of multiple scattering as a function of penetration depth in the medium are analyzed. We also present various considerations for extending this technique to determining structural information in biological tissues, such as the effects of a distribution of particle sizes and the need to average out speckle contributions. PMID- 11934167 TI - Tip/tilt estimation from defocused images. AB - In astronomical imaging, the errors in the wave-front slope are a significant cause of aberrations in the detected image. We investigate how the slope can be estimated optimally using an intensity measurement of the propagated wave front. We show that the optimal location for detection of wave-front tilt is the focal plane, and we quantify the error in using defocused images, such as would be obtained from a curvature sensor, for estimating the wave-front tilt. The effect of using broadband light is also quantified. PMID- 11934168 TI - Bit-error rate for free-space adaptive optics laser communications. AB - An analysis of adaptive optics compensation for atmospheric-turbulence-induced scintillation is presented with the figure of merit being the laser communications bit-error rate. The formulation covers weak, moderate, and strong turbulence; on-off keying; and amplitude-shift keying, over horizontal propagation paths or on a ground-to-space uplink or downlink. The theory shows that under some circumstances the bit-error rate can be improved by a few orders of magnitude with the addition of adaptive optics to compensate for the scintillation. Low-order compensation (less than 40 Zernike modes) appears to be feasible as well as beneficial for reducing the bit-error rate and increasing the throughput of the communication link. PMID- 11934169 TI - Three-dimensional fluorescence enhanced optical tomography using referenced frequency-domain photon migration measurements at emission and excitation wavelengths. AB - The ultimate success of near-infrared optical tomography rests on the precise measurement of light propagation within tissues or random media, the accurate prediction of these measurements from a light propagation model, and an efficient three-dimensional solution of the inverse imaging problem. To date, optical tomography algorithms have focused on frequency-domain photon migration (FDPM) measurements of phase-delay and amplitude attenuation, which are reported relative to the incident light, even though phase-delay and amplitude of incident light are nearly impossible to measure directly. In this contribution, we examine referenced, fluorescence-enhanced frequency-domain photon migration measured at excitation and/or emission wavelengths and report on a measurement strategy to minimize measurement and calibration error for efficient coupling of data to a distorted Born iterative imaging algorithm. We examine three referencing approaches and develop associated inversion algorithms for (1) normalizing detected emission FDPM data to the predicted emission wave arising from a homogeneous medium, (2) referencing detected emission FDPM data to that detected at a reference point, and (3) referencing detected emission FDPM data to detected excitation FDPM data detected at a reference point. Our results show the latter approach to be practical while reducing the nonlinearity of the inverse problem. Finally, in light of our results, we demonstrate the method for eliminating the influence of source strength and instrument functions for effective fluorescence enhanced optical tomography using FDPM. PMID- 11934170 TI - Design of binary long-period fiber grating filters by the inverse-scattering method with genetic algorithm optimization. AB - An approach is presented to the design of binary long-period fiber grating (LPFG) filters based on the Gel'fand-Levitan-Marchenko (GLM) inverse-scattering method and genetic algorithm optimization. The nonuniform coupling strength of the binary grating can be realized by varying the local duty ratio. A coupled-mode theory combined with the Poisson sum formula for treating the binary index perturbation is developed for the application of the GLM synthesis method. Since the coupled-mode theory, which smears out the discrete coupling nature, can be regarded only as an approximation to the modeling of a binary LPFG, we use instead the transfer-matrix model to analyze the coupling behavior of a nonuniform binary LPFG. Based on the synthesized grating patterns from the GLM method, a real-coded genetic algorithm with the transfer-matrix model is used to compensate for the discrepancies resulting from use of the coupled-mode theory and to optimize the design. We exemplify the above procedure by designing a flatband LPFG filter and a high-visibility all-fiber Mach-Zehnder filter. PMID- 11934171 TI - Point-spread function for a rotationally symmetric birefringent lens. AB - Imaging properties of a birefringent lens, in which the fast (or the slow) axis is distributed in the radial direction whereas magnitude of birefringence varies as a quadratic function of the pupil radius, are investigated by calculating a point-spread function. It is found that the point image is analytically described by using the Lommel function as well as the zero-order Bessel function, and a localized intensity null surrounded by bright regions in all directions can be realized at a geometrical focus under certain conditions. The magnitude of birefringence that is tolerable in image formations is also discussed, assuming that the lens is applied to microlithography. PMID- 11934172 TI - Propagation of cylindrically symmetric fields in uniaxial crystals. AB - We investigate the paraxial propagation along the optical axis of a uniaxially anisotropic crystal of a general paraxial beam whose boundary Cartesian components possess cylindrical symmetry. This property allows us to obtain expressions whose dependence on the azimuth angle phi (in cylindrical coordinates) is fully described and very simple. We also find that the beam loses its boundary cylindrical symmetry during propagation, as a consequence of medium anisotropy. Further, these expressions elucidate the way in which the anisotropy changes the state of polarization. As an example, we discuss the case of a Gaussian beam focused into the crystal by a thin spherical lens. PMID- 11934173 TI - Differential interference contrast x-ray microscopy with twin zone plates. AB - X-ray imaging in differential interference contrast (DIC) with submicrometer optical resolution was performed by using a twin zone plate (TZP) setup generating focal spots closely spaced within the TZP spatial resolution of 160 nm. Optical path differences introduced by the sample are recorded by a CCD camera in a standard full-field imaging and by an aperture photodiode in a standard scanning transmission x-ray microscope. Applying this x-ray DIC technique, we demonstrate for both the full-field imaging and scanning x-ray microscope methods a drastic increase in image contrast (approximately 20x) for a low-absorbing specimen, similar to the Nomarski DIC method for visible-light microscopy. PMID- 11934174 TI - Optical properties of an isotropic optically active medium at oblique incidence: comment. AB - The constitutive relations used by Sah and Gowri Krishna [J. Opt. Soc. Am. A 18, 1388 (2001)] are inconsistent with energy and reciprocity requirements. PMID- 11934175 TI - Effect of the polarization on ocular wave aberration measurements. AB - Measurement of the eye's wave aberrations has become fairly standard in recent years. However, most studies have not taken into account the possible influence of the polarization state of light on the wave aberration measurements. The birefringence properties of the eye's optical components, in particular corneal birefringence, can be expected to have an effect on the wave aberration estimates obtained under different states of polarization for the measurement light. In the work described, we used a psychophysical aberrometer (the spatially resolved refractometer) to measure the effect of changes in the polarization state of the illumination light on the eye's wave aberration estimates obtained in a single pass. We find, contrary to our initial expectation, that the polarization state of the measurement light has little influence on the measured wave aberration. For each subject, the differences in wave aberrations across polarization states were of the same order as the variability in aberrations across consecutive estimates of the wave front for the same polarization conditions. PMID- 11934176 TI - The life and times of a good idea. PMID- 11934177 TI - Licensure and membership. PMID- 11934178 TI - Live patients. PMID- 11934179 TI - Mixed messages? PMID- 11934180 TI - Oral cancer campaign. PMID- 11934181 TI - Intra-arterial injection. PMID- 11934182 TI - Antibiotic prescription rates drop. PMID- 11934183 TI - In the aftermath of the Sept. 11 terrorist attacks, have you noted any quantifiable effect on your dental practice? PMID- 11934184 TI - Screening for traditional risk factors for cardiovascular disease: a review for oral health care providers. AB - BACKGROUND: Cardiovascular disease, or CVD, is associated with a very high rate of morbidity and mortality among people in the United States. Primary and secondary prevention measures will help reduce cardiovascular events and increase the overall health of the patient population. METHODS: The author conducted MEDLINE and Internet searches and reviewed publications from professional organizations for the most up-to-date information on CVD and associated risk factors. RESULTS: More than 450,000 articles and monographs on CVD were published in English between January 1990 and May 2001. Of these, approximately 45,000 discussed CVD and risk factors. The author selected and reviewed more than 550 publications on the basis of their relevance to epidemiology, etiology, and primary and secondary prevention of CVD. CONCLUSIONS: Recent information regarding the pathogenesis and treatment of CVD suggests that oral health care providers can perform screening for risk factors and aid in monitoring of specific conditions that put people at an increased risk of developing and exacerbating existing CVD. Dentists need to take a proactive role in this endeavor. CLINICAL IMPLICATIONS: As with most medical conditions, oral health care providers play an important function as it relates to the overall health of their patients. This article provides a timely update on CVD and the relationship between CVD and its risk factors, and offers suggestions for oral health care providers as to when they should interact with patients and patients' primary health care providers to possibly reduce the mortality and morbidity of CVD. PMID- 11934185 TI - Effective treatment of herpes simplex labialis with penciclovir cream: combined results of two trials. AB - BACKGROUND: Two randomized, double-blind, parallel-group clinical trials were conducted in Europe and North America to compare the efficacy and safety of topical 1 percent penciclovir cream with a placebo cream. METHODS: A total of 4,573 immunocompetent people with a history of recurrent herpes simplex labialis, or HSL, with three or more episodes a year that typically manifested as classical lesions, were enrolled and prospectively dispensed medication-either 1 percent penciclovir in a cetomacrogol cream base or a matching placebo. Patients self initiated treatment and were required to apply study medication six times per day for the first day and every two hours while awake for four consecutive days. RESULTS: Of 4,573 enrolled patients, 3,057 initiated treatment (1,516 with penciclovir and 1,541 with placebo). Combined data from two trials revealed that penciclovir recipients lost classical lesions 31 percent faster than did placebo recipients (hazard ratio, or HR, = 1.31; 95 percent confidence interval, or CI, 1.20 to 1.42; P = .0001) and experienced 28 percent faster resolution of lesion pain (HR = 1.28; 95 percent CI, 1.17 to 1.39; P = .0001). Significant benefits were achieved with penciclovir use whether treatment was initiated in the early stages (P = .001) or later stages (P = .0055). CONCLUSIONS: The largest data set currently available on the treatment of recurrent HSL revealed that penciclovir cream significantly outperformed the placebo in healing classical lesions and resolution of pain. CLINICAL IMPLICATIONS: The authors found that penciclovir cream positively affects recurrent HSL, and dose frequency is vital to topical treatment. Even when penciclovir was applied late, it was effective in favorably altering the course of recurrent HSL. PMID- 11934186 TI - The nonsurgical treatment of patients with periodontal disease: results after five years. AB - BACKGROUND: In a previous study involving patients seen at the dental clinic of the Detroit Receiving Hospital, the authors found that 87 percent of teeth initially recommended for surgery or extraction were spared those treatments by a combination of debridement and short-term usage of antimicrobial agents. The objective of the present study was to determine how long the surgery-sparing benefits of less invasive treatment would persist. METHODS: Ninety of these patients were scheduled for maintenance therapy at three-month intervals over a five-year period. They were evaluated periodically for surgical needs by a clinician who was not aware of the nonsurgical periodontal treatment the patient had received. RESULTS: The initial treatment benefits were sustained, as the number of teeth needing periodontal surgery or extraction was 0.06 teeth per patient after 1.1 year, 0.22 after 2.3 years, 0.51 after 3.6 years and 0.86 after 5.1 years. CONCLUSIONS: A noninvasive treatment regimen for an anaerobic infection in teeth seriously compromised by periodontal disease resulted in a reduced need for surgery or tooth extraction for at least five years after completion of the initial treatment. PMID- 11934187 TI - Women's health issues and their relationship to periodontitis. AB - BACKGROUND: The emergence of sex-specific associations between periodontitis and certain systemic disorders has prompted researchers to investigate the possibility of associations between periodontitis and specific women's health issues. The authors review the potential relationships between periodontitis and hormonal changes and their ramifications in regard to pregnancy outcomes, cardiovascular disease, or CVD, and osteoporosis. METHODS: Changes in hormone levels, such as those that occur during puberty, pregnancy, menstruation and menopause, as well as those that occur with the use of hormonal supplements, have long been associated with the development of gingivitis. Furthermore, bacterial anaerobes have been found to change during the normal hormonal cycle. In periodontitis, the inflammatory response results in ulceration of the gingivae and the subsequent entry of bacterial cells, bacterial products, peptidoglycan fragments and hydrolytic enzymes into the systemic circulation. The result is a systemic response of increased cytokines and biological mediators, as well as increased levels of serum antibodies. RESULTS: Some researchers have found that pregnant women with periodontitis were 7.5 times more likely to have a preterm low-birth-weight infant than were control subjects. Other researchers reported that the risk of preterm birth was directly related to the severity of periodontitis. Similarly, researchers have linked periodontitis to CVD. Many studies have indicated that estrogen exerts a protective effect against CVD development, and much evidence suggests that when hormone replacement therapy is administered to postmenopausal women, this effect continues. A relationship between periodontitis and osteoporosis has been established, such that more clinical attachment loss has been noted in osteoporotic people. CONCLUSIONS: The literature suggests that more sex-specific research is essential to determine the strategies needed to prevent and treat adverse pregnancy outcomes, CVD and osteoporosis through hormone modification and periodontitis control. CLINICAL IMPLICATIONS: Dentists must assume greater responsibility for the overall health of their patients, and acquire knowledge of relevant systemic conditions to interact meaningfully with medical colleagues. PMID- 11934188 TI - A technique to enhance closed surgical stents for implant placement. PMID- 11934189 TI - A comparison of polymerization by light-emitting diode and halogen-based light curing units. AB - BACKGROUND: Light-emitting diode, or LED, technology provides certain advantages over halogen-based light polymerization of resin-based composites. The authors investigated the adequacy of cure of LED light-curing units, or LCUs. METHODS: The authors used two halogen-based light-curing units (Optilux 400 and 501, Demetron Research Corp., Danbury, Conn.) and two commercially available LED LCUs (LumaCure, LumaLite, Spring Valley, Calif., and VersaLux, Centrix, Shelton, Conn.) to polymerize top surfaces of hybrid (Filtek Z-250, 3M, St. Paul, Minn.) and microfilled (Renamel, Cosmedent, Chicago) resin-based composite specimens. Specimens were indented on their top and bottom surfaces with a Knoop hardness tester and measured for hardness. Bottom:top hardness ratios determined the percentage of cure. The authors separated the data into eight groups (two composites cured with four different lights) with 15 observations per group, for a total sample size of 120. RESULTS: The authors compared composites and curing lights by a two-way analysis of variance, and results indicated significant main effects. The main effect of composite was statistically significant (P < .0001) when microfilled composite was compared with hybrid composite, regardless of curing light, for all top and bottom hardness measurements, with the hybrid producing much higher hardness measurements overall. The main effect of light was significant as well (P < .0001), regardless of composite type, with the two halogen-based lights producing harder top and bottom composite surfaces than the two LED LCUs. CONCLUSIONS: The light output of commercially available diodes for resin-based composite polymerization still requires improvement to rival the adequacy of cure of halogen-based LCUs. Additional studies are necessary. CLINICAL IMPLICATIONS: Commercially available LED LCUs were introduced just in the past year. However, they may not adequately polymerize resin-based composites, which can lead to restoration failures and adverse pulpal responses to unpolymerized monomers. PMID- 11934190 TI - Patterns of oral care in a Washington State dental service population. AB - BACKGROUND: The authors compare patterns of oral health care reported by the Washington Dental Service, or WDS, Seattle, in 1993 and 1999 to assess changes in patient populations, practice characteristics, procedures and treatment costs in the state. METHODS: Data were obtained from dental benefits claims from a population of about 1.25 million people. Variables of interest included patient age and other demographic information, character of dental practice, dental procedures and treatment costs that combined WDS payment and patient copayment. RESULTS: The results showed high agreement (97 percent) between the database and randomly surveyed patient records. For both 1993 and 1999, general dental offices were responsible for more than 80 percent of patient care. Single crowns (21 percent), restorative services (15 percent) and dental prophylaxis (13 percent) made up about half of the costs of dental care. Broad categories of service were similar in 1993 and 1999, and anticipated major declines in restorative procedures related to caries were not apparent. The mix of services varied considerably by patient age and between generalists and specialists in both years. CONCLUSIONS: Patterns of oral health care among this insured patient population largely remained unchanged from 1993 to 1999, with some shifts in specific procedures and specialty care. During this period, dentists saw more patients and performed fewer treatments per patient, while total treatment costs per patient increased. CLINICAL IMPLICATIONS: Patterns of oral health care in the United States are projected to undergo major changes linked to improved oral health, declining trends in caries and periodontal diseases, scientific advances in treatment approaches and a patient population that is living longer. Changes in care patterns during this six-year period may reflect patient and provider preferences, as well as the influence of reimbursement policies. Dental benefits databases can serve as a critical resource for monitoring such changes. PMID- 11934191 TI - Computer-assisted analysis of oral brush biopsies at an oral cancer screening program. AB - BACKGROUND: Dentists and dental hygienists attending a health screening program were screened for oral cancer. Select oral epithelial lesions were evaluated by oral brush biopsy with a computer-assisted method of analysis (OralCDx, OralScan Laboratories Inc., Suffern, N.Y.). METHODS: After those who had oral epithelial lesions were identified, the clinical characteristics of each lesion were recorded. Participants with abnormal oral brush biopsy results ("positive" and "atypical") subsequently underwent incisional biopsy of their lesions by an oral surgeon. RESULTS: A total of 930 dentists and dental hygienists were screened over a four-day period at each of the American Dental Association's 1999 and 2000 annual sessions. Eighty-nine people (9.7 percent) with 93 oral epithelial lesions were identified and evaluated by brush biopsy. Seven of the 93 oral lesions-all benign appearing in their clinical appearance-were determined to be "atypical" or "positive." Of these, three were diagnosed as precancerous by scalpel biopsy and histologic evaluation. CONCLUSIONS: Computer-assisted brush biopsy analysis is a valuable adjunct to the oral screening examination. The identification of three innocuous-looking precancerous lesions in this low-risk group of dentists and dental hygienists underscores the necessity of evaluating all oral lesions of unknown etiology. CLINICAL IMPLICATIONS: As 9.7 percent of the screened dentists and dental hygienists had epithelial oral lesions, general dentists most likely routinely encounter an even higher percentage of oral lesions in their patients. The minimally invasive brush biopsy lets general dentists evaluate these lesions. Like Pap smears and mammograms, this tool can help identify precancers and potentially curable cancers. PMID- 11934192 TI - Office emergencies and emergency kits. AB - Preparedness to recognize and appropriately manage medical emergencies in the dental environment includes the following: current BLS certification for all office staff; didactic and clinical courses in emergency medicine; periodic office emergency drills; telephone numbers of EMS or other appropriately trained health care providers; emergency drug kit and equipment, and the knowledge to properly use all items. This update on office emergencies and emergency kits replaces the previous Association report, prepared in 1980 by the Council on Dental Therapeutics. PMID- 11934193 TI - Orthodontics and the general practitioner. AB - Until now, general dentists have not had significant involvement with orthodontic procedures. With this article, I encourage general dentists and other nonorthodontists interested in orthodontic therapy to acquire adequate education to allow them to participate in this area of clinical activity. The Invisalign concept can enhance the desirability of nonorthodontists' becoming involved with orthodontic procedures. PMID- 11934194 TI - HIV and discrimination: a review of the Waddell case and its implications for health care professionals. PMID- 11934195 TI - For the dental patient. Common mouth sores and patches. PMID- 11934196 TI - Immediate transepithelial photorefractive keratectomy for treatment of laser in situ keratomileusis flap complications. AB - PURPOSE: To study the role of photorefractive keratectomy (PRK) in the management of laser in situ keratomileusis (LASIK) flap complications. METHODS: A retrospective analysis was performed of seven patients (14 eyes) who had bilateral, simultaneous LASIK (single surgeon [TGA], Summit Apex Plus laser, Hansatome microkeratome in six patients [12 eyes], Automated Corneal Shaper keratome in one patient [two eyes]). One eye of each patient received PRK for a flap complication. The seven patients (four women, three men) had a mean age of 39.2 years (range, 22 to 64 yr). The uncomplicated LASIK eyes had moderate to high myopia (-2.25 to -8.75 D) and the eyes with LASIK flap complications had mild to moderate myopia (-1.50 to -5.75 D). Immediate transepithelial PRK was performed in all eyes after repositioning the defective flap. RESULTS: All seven patients had excellent visual outcome at 6 months follow-up; one patient required an enhancement (astigmatic keratotomy). Six of the seven LASIK eyes had best spectacle-corrected visual acuity of 20/20 or better and one LASIK eye had 20/25. In the transepithelial PRK eyes, six of the seven eyes had best spectacle corrected visual acuity of 20/20 or better and one eye had 20/30. CONCLUSIONS: Immediate transepithelial PRK treatment of irregular and incomplete LASIK flaps at the time of surgery is an excellent therapeutic option to delayed surgical treatment. PMID- 11934197 TI - Effect of laser in situ keratomileusis for hyperopia on tear film and ocular surface. AB - PURPOSE: To examine the effects of laser in situ keratomileusis (LASIK) for hyperopia on the tear film and ocular surface. METHODS: A retrospective 12-month analysis of 88 eyes (88 participants) who had LASIK for hyperopia was performed. Participants were evaluated before and after (2 weeks, 1, 3, 6, and 12 months) surgery for dry eye symptoms (McMonnies Dry Eye Survey primary symptoms), tear film stability (fluorescein break-up time), tear volume (phenol red thread test), ocular surface staining (fluorescein), and conjunctival goblet cell density. RESULTS: Chronic dry eye was experienced by 32% of participants; symptoms were significantly associated with female gender, preoperative dry eye symptoms, lower tear film stability after surgery, greater ocular surface staining after surgery, lower tear volume before and after surgery, and lower goblet cell densities after surgery. Regression rate 12 months after surgery was 32% and significantly associated with female gender, chronic dry eye symptoms, lower tear film stability after surgery, greater ocular surface staining before and after surgery, and lower tear volume before and after surgery. CONCLUSIONS: Dry eye, particularly in females, is problematic after LASIK for hyperopia and is associated with refractive regression. Current methods for managing the tear film and ocular surface may not control LASIK-induced dry eye, particularly in some females during the first 6 months after surgery. PMID- 11934198 TI - Tear secretion following excimer laser in situ keratomileusis. AB - PURPOSE: To evaluate the effect of laser in situ keratomileusis (LASIK) on tear secretion. METHODS: A non-consecutive series of 42 eyes of 42 patients had LASIK for the correction of myopia and myopic astigmatism. The fellow eye served as a control. The mean preoperative spherical equivalent refraction was -5.25 +/- 1.00 D (range, -1.75 to -11.00 D). Attempted correction aimed at emmetropia. Schirmer I and II, and tear break-up time (BUT) tests were performed preoperatively and at 1, 3, and 6 months postoperatively. All tests were correlated to the amount of the attempted correction. RESULTS: Preoperatively, mean values were Schirmer I: 16.2 mm, Schirmer II: 11.6 mm, and BUT: 16.3 sec. One month after LASIK, mean values were Schirmer I: 12.8 mm, Schirmer II: 9.2 mm, and BUT: 13.3 sec. At 3 months, mean values were Schirmer I: 15.76 mm, Schirmer II: 11.3 mm, and BUT: 14.15 sec. At 6 months, mean values were Schirmer I: 15.96 mm, Schirmer II: 11.66 mm, and BUT: 16.3 sec. No correlation to the amount of attempted correction was found. Fellow eye tests were not affected at any interval. CONCLUSIONS: Tear secretion following LASIK was decreased during 3 months after surgery and was normalized by 6 months. PMID- 11934199 TI - Evaluation of tear film following Intacs for myopia. AB - PURPOSE: To evaluate the effect of intrastromal corneal ring segments (Intacs, Keravision) placement on the tear film quality as a measure of dry eye status. METHODS: Seventeen eyes of ten patients underwent Intacs placement for the correction of myopia with spherical equivalent refractions ranging from -1.75 to 3.25 D (mean, -2.43 +/- 0.56 D). Dry eye grade values were measured preoperatively and at 1 day, 1 week, and 1 month postoperatively using the Kowa DR-1 interferometer. RESULTS: Preoperatively, the 17 eyes had a mean dry eye grade of 2.18 +/- 0.39 (range, 2 to 3). One day postoperatively, the 14 eyes measured had a mean dry eye grade of 2.64 +/- 0.63 (range, 2 to 4; P = .03). At 1 week the 17 eyes had a mean dry eye grade of 2.41 +/- 0.71 (range, 2 to 4; P = .25) and at 1 month, 2.18 +/- 0.39 (range, 2 to 3; P = .75). CONCLUSIONS: There was transient dry eye following Intacs placement, but the tear film quality was restored within 1 week of surgery. PMID- 11934200 TI - Comparison of TGF-beta1 in tears following laser subepithelial keratomileusis and photorefractive keratectomy. AB - PURPOSE: To compare the release of tear fluid transforming growth factor-beta1 (TGF-beta1) of 15 patients who underwent photorefractive keratectomy (PRK) in one eye and laser subepithelial keratomileusis (LASEK) in the other eye. METHODS: Tear fluid samples were collected with scaled microcapillary tubes preoperatively (day 0) and on the first, second, and seventh postoperative days. We calculated the release of TGF-beta1 (tear fluid flow-corrected concentrations of TGF-beta1) by multiplying the concentration by capillary tear fluid flow and also evaluated corneal haze at 1, 3, and 6 months after PRK or LASEK. RESULTS: The median TGF beta1 release values were: Day 0: 50.6 pg/min (range 0 to 101.6 pg/min) for PRK and 56.7 pg/min (range 0 to 121.8 pg/min) for LASEK (P=.496); Day 1: 240.6 pg/min (range 56.6 to 1120.2 pg/min) for PRK and 155.2 pg/min (range 45.1 to 480.4 pg/min) for LASEK, which showed a significant difference between the two procedures (P=.041); Day 2: 395.7 pg/min (range 92.3 to 1150.7 pg/min) for PRK and 185.2 pg/min (range 41.4 to 872.5 pg/min) for LASEK (P=.004); and Day 7: 91.3 pg/min (range 0 to 225.4 pg/min) for PRK and 74.3 pg/min (range 0 to 187.1 pg/min) for LASEK, (P=.244). The mean corneal haze score at 1 month after PRK was 0.88 +/- 0.50 and 0.53 +/- 0.29 after LASEK, and a statistically significant difference was noted between the two groups (P=.005). At 3 months (P=.083) and 6 months (P=.157) after the procedures, there were no statistically significant differences. CONCLUSIONS: A lower amount of tear fluid transforming growth factor beta1 was released in the early postoperative days following LASEK than in PRK. The lower grade of corneal haze seen in LASEK than PRK in the early postoperative period may come from a decreased release of TGF-beta1, which may have a role in wound healing. PMID- 11934201 TI - Concomitant effect of topical ubiquinone Q10 and vitamin E to prevent keratocyte apoptosis after excimer laser photoablation in rabbits. AB - PURPOSE: To investigate in vivo whether ubiquinone Q10 together with vitamin E protects rabbit corneas from keratocyte apoptosis after excimer laser irradiation. METHODS: Photorefractive keratectomy (PRK) was performed in both eyes of three New Zealand white rabbits. During 3 days before surgery, each right eye received four-times-daily instillation of an eye-drop solution containing ubiquinone Q10 0.20% and vitamin E 0.04%; each left eye was treated with a solution that did not contain ubiquinone or vitamin E. The central cornea was analyzed after surgery using the in situ end labelling (ISEL) technique of nicked DNA to detect DNA fragmentation. To determine the number of ISEL positive nuclei, an average of 70 random microscopic fields (five for each de-epithelialized tissue section) of 138,000 mu2 were examined in the right and left cornea samples at 250X by two different observers. RESULTS: Light microscopic examination of the sections from corneas treated before PRK showed that cells committed to apoptosis by PRK were about 50% compared to those of untreated controls. CONCLUSION: Treatment of rabbit eyes before PRK with ubiquinone Q10 lowered the number of apoptotic events. PMID- 11934203 TI - Effect of topical diclofenac and ketorolac on patient discomfort and corneal sensitivity. AB - PURPOSE: To determine if at clinical dosages, the topical nonsteroidal anti inflammatory drugs diclofenac and ketorolac decrease corneal sensitivity, and if there is a difference in discomfort on instillation. METHODS: In a double-masked study, 30 patients were randomized to receive two of three topical drops, Endosol, diclofenac sodium, and ketorolac tromethamine. A microaesthesiometer was used to evaluate corneal sensation. Baseline corneal sensitivity was obtained on each patient. Corneal sensory thresholds were remeasured at 7 and 15 minutes following eyedrop application. Patients also completed a questionnaire that graded the burning caused by the medications. RESULTS: No significant decrease in corneal sensory thresholds was found with diclofenac or ketorolac compared with control at baseline (P = .50), 7 minutes (P = .41), or 15 minutes (P = .82). There was significantly more burning with diclofenac and ketorolac compared to control. There was a small but not statistically significant (P = .28) trend of more burning with ketorolac than diclofenac. CONCLUSION: Neither diclofenac nor ketorolac were found to decrease corneal sensation compared to control. There was no significant difference in burning upon instillation reported by patients. PMID- 11934202 TI - Pain reduction after laser in situ keratomileusis with ketorolac tromethamine ophthalmic solution 0.5%: a randomized, double-masked, placebo-controlled trial. AB - PURPOSE: To evaluate the analgesic efficacy of ketorolac tromethamine ophthalmic solution 0.5% after laser in situ keratomileusis (LASIK). METHODS: In this two center, randomized, double-masked, placebo-controlled, parallel group study, 39 patients underwent bilateral simultaneous LASIK. Patients received study drops (Acular PF or Lens Plus) in both eyes 15 to 30 minutes before surgery, again immediately before passing of the microkeratome, and again after flap repositioning. Proparacaine was used during surgery, but no additional therapeutics were used for the next 24 hours, except acetaminophen or propoxyphene napsylate acetaminophen allowed as escape medication. Patients rated their eye pain hourly through 6 hours after surgery. RESULTS: Ketorolac significantly reduced eye pain at every time point compared to placebo (P<.01). Escape medication use declined significantly; 16% (3/19) of those who received ketorolac required escape medication compared to 50% (8/16) of placebo-treated patients (P=.03). Ketorolac-treated eyes were pain-free significantly sooner (P<.01), with 47% (18/38) having pain cessation by hour 4, compared to 15% (5/33) of placebo-treated eyes. No treatment-related adverse events occurred. CONCLUSION: This study supports the use of topical ketorolac for control of early postoperative pain following LASIK, significantly increasing patient comfort and reducing usage of other pain medications. PMID- 11934204 TI - Flap measurements with the Hansatome microkeratome. AB - PURPOSE: To evaluate flap thickness, flap diameter, and hinge length during laser in situ keratomileusis (LASIK) and to correlate these measurements with preoperative keratometric power, central corneal thickness, and patient refraction, gender, and age. METHODS: In this prospective study of 50 eyes of 28 patients (mean age 31 +/- 6.6 yr; range, 24 to 43 yr) results of LASIK for myopia were analyzed (mean spherical equivalent refraction of -7.16 +/- 1.69 D; range, 2.75 to -13.50 D). Corneal flaps were created using the Hansatome microkeratome (Baush & Lomb Surgical) with a 160-microm plate and a 9.5-mm suction ring. Corneal thickness was evaluated using an ultrasonic 50-MHz pachymeter (Sonogage Corneo Gage Plus) and the mean keratometric power was measured with a Corneal Analysis System videokeratographic unit (EyeSys). Data were analyzed using t test, Pearson product moment correlation coefficient, and Spearman's rho non parametric correlation coefficients. RESULTS: Mean corneal flap thickness was 142.6 +/- 20.8 microm (range, 107 to 177 microm), mean flap diameter was 9.9 +/- 0.3 mm (range, 9.2 to 10.5 mm), and mean hinge length was 6.2 +/- 0.4 mm (range, 5.2 to 7 mm). Statistically significant correlations (P<.05) were found between mean keratometric power and flap hinge length, mean keratometric power and flap diameter, preoperative spherical equivalent refraction and flap diameter, corneal thickness and flap hinge length, as well as patient age and corneal thickness. CONCLUSIONS: The Hansatome microkeratome was an effective and safe instrument in the creation of corneal flaps for LASIK. Consideration of preoperative keratometric power and corneal thickness may help to reduce or avoid complications. PMID- 11934205 TI - Repeated photorefractive keratectomy for undercorrection and regression. AB - PURPOSE: Regression (often combined with significant haze) and undercorrection are the most frequent complications after photorefractive keratectomy (PRK) for myopia. PRK retreatment has been used to treat both of these complications. METHODS: Sixty-three eyes (55 patients) were reoperated (PRK) because of initial regression or undercorrection. The mean interval between the operations was 17.2 months (range 5 to 37 mo). Residual myopia before retreatment was less than -5.00 D in 62 eyes (mean -2.21 +/- 0.99 D) and was -10.50 D in one eye. The same surgical PRK technique (Aesculap-Meditec MEL 60 excimer laser with 5 or 6-mm ablation zone) was used in both operations. RESULTS: One-year results were available for 51 eyes. In the group of eyes with low residual myopia (<-5.00 D), mean refraction 1 year after retreatment was -0.42 +/- 0.96 D and 43 eyes (86%) were within +/-1.00 D of emmetropia. Uncorrected visual acuity of 20/40 or better was achieved in 43 eyes (86%) after 1 year. One eye lost two lines of best spectacle-corrected visual acuity after two procedures because of haze. Mean haze was the same before and 12 months after reoperation (grade 0.39 vs. 0.38). Significant haze (>2) and high regression after the first PRK occurred after reoperation. The one eye with -10.50 D residual myopia after the first PRK showed high regression, and 1 year after the reoperation refraction was -9.50 D with grade 2 haze. CONCLUSIONS: Repeated PRK was safe and in most cases effective in treating regression and undercorrection in eyes with low residual myopia after initial PRK. High regression and especially haze after the initial PRK often reappeared after reoperation. PMID- 11934206 TI - Cost-effectiveness of the AMOArray multifocal intraocular lens in cataract surgery. AB - PURPOSE: To compare the cost and outcomes of bilateral cataract surgery with the foldable AMOArray multifocal intraocular lens (MIOL) versus the foldable monofocal intraocular lens from the health care payer perspective. METHODS: A cost-effectiveness analysis was used to evaluate treatment with the MIOL compared to monofocal lens, using data from clinical trials, literature, expert opinion, and a review of the German health care funding and reimbursement system. RESULTS: The average total direct medical cost per patient (per procedure) with the MIOL was DM 1,774 compared to DM 1,716 with the monofocal lens (1DM = US$0.558 in April 1998). The MIOL was more cost-effective than the monofocal lens in terms of cost per patient (spectacle-free). Cost per patient without overall limitation in vision-related function and cost per patient without limited night vision were similar for both patient groups. The incremental cost of the MIOL for a one-point increase was DM 52 in the self-rated score "quality of vision," DM 82 in "satisfaction with day vision," and DM 115 in "satisfaction with night vision." CONCLUSION: The small additional cost of the MIOL was outweighed by the increased satisfaction with vision experienced by MIOL patients. PMID- 11934207 TI - Comparison of Placido-based, rasterstereography, and slit-scan corneal topography systems. AB - PURPOSE: Elevation-based topography systems have received growing recognition. We compared a Placido-based (EyeSys), a rasterstereography (PAR), and a slit-scan system (Orbscan) for human subjects. METHODS: Measurements were obtained from 221 eyes of 119 human subjects. We made statistical comparisons central curvature, keratometric curvatures, and meridians between the three systems. We also compared the optical pachymetry of the slit-scan system with an ultrasonic device. We analyzed cases of subclinical keratoconus, central island, photorefractive keratectomy for myopia, and hyperopia correction with arcuate keratotomy. RESULTS: The statistical comparison showed better correlation between the Placido-based and the rasterstereography systems in measurement of central curvature (R=0.95). The slit-scan system showed better correlation with the Placido system in the preoperative and postoperative radial keratotomy group (R=0.73) than in the group that included postoperative PRK eyes (R=0.69). Similar results were obtained for keratometric curvatures and meridians. The Orbscan pachymetry correlation with ultrasonic was R=0.69 and increased when the postoperative PRK eyes were excluded (R=0.95). CONCLUSIONS: Slit-projection topography and pachymetry seemed to be affected by hydration in the postoperative PRK group. The pachymetric map was useful in predicting the final outcome of refractive surgery and in the diagnosis of keratoconus. PMID- 11934208 TI - Early onset ectasia following laser in situ keratomileusus: case report and literature review. AB - PURPOSE: Laser in situ keratomileusis (LASIK) has been associated with the development of postoperative corneal ectasia. We present a case of early onset ectasia after LASIK, review known risk factors in development, and discuss possible strategies for prevention. METHODS: A 39-year-old man underwent bilateral LASIK for moderate myopia. Preoperative cycloplegic refractions were 9.00 + 0.25 x 140 degrees OD and -7.75 sphere OS. Corneal topography demonstrated mild inferior steepening bilaterally although definite evidence of keratoconus by either the Klyce/Maeda and Smolek/Klyce keratoconus screening tests was not present. Following the creation of flaps with 160-microm plates, ablations of 93 microm OD and 80 microm OS were performed, estimated to leave residual stromal beds of at least 314 microm OD and 330 microm OS. RESULTS: On the first postoperative day, uncorrected visual acuities were 20/400 OD and 20/40 OS. On the fifth postoperative day, the patient's uncorrected visual acuity was 20/400 OD, and 20/300 OS. Corneal topography of the right eye showed profound inferior steepening with an apical corneal power in excess of 57 D; topography of the left eye showed mild inferior steepening. Eighteen months after surgery best corrected visual acuity was 20/40 OD and 20/30 OS with rigid gas permeable contact lenses. CONCLUSIONS: This case highlights the need for a high index of suspicion when one notes an asymmetric bow-tie pattern on preoperative LASIK corneal topography, despite seemingly safe estimates of residual stromal bed thickness. PMID- 11934209 TI - U.S. trends in refractive surgery: 2001 International Society of Refractive Surgery Survey. AB - PURPOSE: To determine the trends in refractive surgery in the United States in 2001. METHODS: The 980 U.S. members of the International Society of Refractive Surgery were mailed the 2001 refractive surgery survey dealing with volumes, types, and preferences of refractive surgery performed. Questions regarding radial keratotomy, astigmatic keratotomy, photorefractive keratectomy, laser in situ keratomileusis (LASIK), laser subepithelial keratomileusis, intracorneal rings (Intacs), laser thermal keratoplasty, conductive keratoplasty, clear lens extraction, phakic intraocular lenses, and scleral expansion procedures for presbyopia were examined in the survey. Procedure preferences for low, moderate, and high myopia and hyperopia were compared with the results from the surveys of the previous 4 years. Preference for unilateral versus bilateral same-day surgery, laser type, and microkeratome choice were also compared with the survey data from previous years. Incidence and frequency of comanagement of refractive surgery patients were compared with 2000 data. RESULTS AND CONCLUSION: LASIK remains the dominant refractive surgery for refractive errors from -12.00 to +3.00 D; the VISX excimer laser and the Hansatome microkeratome are the most frequently used instruments for LASIK. PMID- 11934210 TI - Correlation of pupil size to iris color in children. PMID- 11934211 TI - Oral heparins. AB - The antithrombotic drug heparin is administered parenterally and believed not effective orally. Oral heparin would be most suitable for long term administration, often required for the prevention of thrombosis. Following parenteral administration, heparin is taken up by endothelial cells. Our laboratory has shown that heparin is similarly taken up by endothelium following oral administration, despite low plasma heparin concentrations. In a twenty-four hour period, endothelial heparin concentrations are greatest within 15 minutes of oral dosing although plasma levels never exceed one percent of dose. Endothelial uptake accounts for a considerable amount of absorption if the total body endothelium is considered. In support of oral heparin absorption, we demonstrated a dose-dependent decrease in thrombosis incidence in a rat jugular vein model following single oral doses of unfractionated heparins (bovine and porcine) or low molecular weight heparins (reviparin, logiparin and ardeparin). Low molecular weight heparins were effective at lower doses than unfractionated heparins where a fifty percent reduction in thrombosis was observed with 0.025 mg/kg reviparin, 0.1 mg/kg logiparin, versus 7.5 mg/kg bovine unfractionated heparin. These studies support the work of others demonstrating measurable systemic changes following oral heparin administration and suggest that heparin may be effective when administered by the oral route. It also indicates that the presence of heparin in plasma likely reflects a much greater amount associated with endothelium. PMID- 11934212 TI - Analysis of commercial CD34 control samples for quality assurance. AB - The amount of CD34+ cells is important to predict the quality of stem cell transplants and ensure safe engraftment after high-dose chemotherapy. Further, daily controls of the patients' peripheral blood CD34+ cell counts are performed to optimize peripheral blood stem cell collection, especially in patients with low CD34+ cell numbers. Therefore, the use of reliable reference samples is mandatory for quality assurance, both in terms of patient safety and reproducibility of results from different laboratories. We report our first experience with CD-Chex CD34, containing stabilized placental cord blood cells in preservative medium with defined concentrations of CD34+ cells. Analysis was performed according to standard operating procedures. Three lots were tested sequentially on a day per day use. The expected CD34+ values were 28.6-, 36.7- and 28.1 microl(-1), respectively, and the mean measured values were 27.4 +/- 2.75 microl(-1) (n = 25, range 21 - 33 microl(-1), coefficient of variation [CV] 10.0%), 29.9 +/- 2.39 microl(-1) (n = 17, range 26 - 35 microl(-1), CV 8.0%), and 27.2 +/- 2.24 micro1(-1) (n = 18, range 24 - 34 microl(-1), CV 8.2%). Serial dilution (1:2 to 1:10) with normal peripheral blood or PBS w/o Ca++/Mg++ gave adequate results. We conclude that the control samples used in this setting are reliable and thus helpful to increase the accuracy in the analysis of CD34+ cells. PMID- 11934213 TI - Plasminogen activator inhibitor in patients with acute myocardial infarction and re-infarction in syndrome X. AB - Plasminogen activator inhibitor was determined in myocardial infarction and re infarction patients along with other parameters relevant for the Syndrome X, such as triglycerides, HDL-cholesterol and body mass index. Plasminogen activator inhibitor was determined in 58 patients with myocardial infarction and 18 patients with re-infarction as well as in 57 healthy individuals. Significantly higher plasminogen activator inhibitor values were found in re-infarction patients (6.53 +/- 0.39 U/mL) than in myocardial infarction patients (4.32 +/- 0.19 U/mL), whereas the values were significantly higher in both groups in comparison to healthy individuals (2.81 +/- 0.74 U/mL). Triglyceride values were also significantly higher in myocardial infarction (2.05 +/- 0.1 mmmi/L) and re infarction patients (2.02 +/- 0.99 mmol/L) than in healthy individuals (1.35 +/- 0.58 mmol/L), although the difference between the triglyceride values obtained for myocardial infarction and re-infarction patients was not statistically significant. HDL-cholesterol values for myocardial infarction (1.18 +/- 0.04 mmol/L) and re-infarction (1.14 +/- 0.18 mmol/L) patients were significantly lower than in healthy individuals (1.46 +/- 0.5 mmol/L). However, the difference was not statistically significant between the myocardial infarction and the re infarction group. The difference in body mass index between myocardial infarction (26.65 +/- 0.39 kg/m2) and re-infarction (25.49 +/- 0.39 kg/m2) patients and healthy individuals (24.45 +/- 3.41 kg/m2) was evidenced. PMID- 11934214 TI - The role of low levels of the serum glutathione-dependent peroxidase and glutathione and high levels of serum homocysteine in the development of cardiovascular disease. PMID- 11934215 TI - Multicenter evaluation of a new immunoassay for intact PTH measurement on the Elecsys System 2010 and 1010. AB - BACKGROUND AND OBJECTIVE: The determination of parathyroid hormone (PTH) is of great clinical relevance in the assessment of calcium metabolic disorders. Although PTH was one of the first hormones measured by immunoassays, there are still many difficulties in its determination due to the low concentration of the hormone in blood and due to the heterogeneity of PTH resulting from different circulating hormone fragments. The aim of our multicenter-study was to evaluate the technical performance and the clinical validity of a new immunoassay for intact PTH measurement on the Elecsys Systems 2010 and 1010. METHODS AND RESULTS: The multicenter evaluation was performed in 11 clinical laboratories. The Elecsys PTH assay is a one step sandwich electrochemiluminescence immunoassay based upon the streptavidin-biotin technology. Two monoclonal antibodies are used in the assay providing detection of intact PTH. The imprecision study yielded within-run and between-days coefficients of variation of 3.1% - 6.6% and 3.4% - 15.6%, respectively using a three level control (PreciControl Bone, Roche Diagnostics) and human pool sera at two different concentrations (HS-low: 20 - 60 pg/ml, HS high > 65 pg/ml). The analytical sensitivity calculated as the mean value plus 2 standard deviations of a within-run imprecision was below 2.70 pg/ml using zero calibrator matrix. Dilution linearity was observed up to 4890 pg/ml using zero calibrator matrix or human pool sera. Recoveries ranged between 85% - 115%. Serum, EDTA- and heparin plasma were evaluated for PTH measurement. Due to a better analyte stability (48h at 21 degrees C; 3d at 4 degrees C) EDTA plasma was recommended for PTH measurement. Results of the Elec sys PTH immunoassay correlated well (r = 0.926 - 0.994) with three different immunoradiometric assays (N-tact PTH SP, DiaSorin; Nichols Allegro Intact PTH, Nichols Institute Diagnostics; ELSA-PTH, CISBio International) and two different immunochemiluminometric assays (PTH-Intact-Immulite, DPC Biermann; Nichols Advantage Intact PTH, Nichols Institute Diagnostics) in technical and clinical method comparisons. The Passing/Bablok regression analysis yielded slopes of 0.692 - 1.729 and intercepts of -13.982 - +15.763 pg/ml. Deviations from slope 1.0 and intercept 0.0 were not unexpected due to differences in immunoassay standardization and probably due to the presence of different PTH fragments and a variable affinity of the used antibodies to these PTH fragments. Highly similar PTH concentration pattern of the Elecsys immunoassay and the Quick-Intraoperative Intact PTH immunoassay (Nichols Institute Diagnostics) obtained from specimens taken intraoperatively support the applicability of the Elecsys immunoassay to monitor the success of parathyroid resection. A reference range of 12.3 - 56.0 pg/ml calculated from PTH values of 43 apparently healthy individuals confirms reference limits published in the literature. The partition of collectives according to age showed, that individuals > 50 years have slightly higher PTH concentrations, independently of gender. This shift could be due to age itself or to an increased prevalence of individuals without obvious calcium metabolic disorders in this collective. CONCLUSION: The Elecsys PTH assay is a useful and reliable tool for determination of intact PTH. Our data support the intended use of the assay in clinical applications related to disorders of calcium metabolism. PMID- 11934216 TI - Oral glucose tolerance test: to be or not to be performed? AB - Type 2 diabetes is one of the most common diseases worldwide. In addition, there are data for a large cohort of undiagnosed cases. In the RIAD (Risk factors in Impaired glucose tolerance for Atherosclerosis and Diabetes) study a total of 15.1% of so far undiagnosed diabetic subjects were detected as well as 26% of subjects with impaired glucose tolerance in a German risk population for diabetes. Type 2 diabetes is associated with an excessively high mortality and morbidity due to cardiovascular disease. Numerous studies have demonstrated the relevance of postprandial hyperglycemia for atherosclerosis. Moreover, the form of isolated postprandial diabetes seems to be much more common than expected. Even mild postprandial hyperglycemia in the form of impaired glucose tolerance was shown to be associated with an increased rate of cardiovascular disease. This indicates the necessity of using the oral glucose tolerance test (OGTT) in the screening of high-risk populations in order to detect asymptomatic diabetic subjects and enable appropriate treatment in time. Not using the OGTT would mean missing a large cohort of undiagnosed diabetic subjects, particularly among older people. Since an OGTT cannot be generally conducted, we recommend its performance in risk subjects and especially in elderly women. This would make it possible to institute preventive measures. PMID- 11934217 TI - The use of nucleic acid amplification techniques to increase the viral safety of blood. AB - Despite recent significant improvements in the viral safety of blood and blood products there remains a small risk of contamination mainly due to the existence of a window period before the appearance of antibodies. Nucleic acid amplification technologies (NAT) permit a direct detection of the viral genome itself with an extreme sensitivity and specificity, without depending anymore on the delayed appearance of antibodies. These technologies can be applied to detect most blood-borne viruses. However, the usefulness and strategies will largely depend on different features specific to the type of the virus, such as pathogenicity, prevalence of the infection, viral load during preseroconversion, doubling time of the virus and infectious dose. Many studies have already been conducted in different parts of the world, and the results proving the feasibility of the NAT screening are more than encouraging. However, some problems still remain to be solved in theroutine application of these technologies. PMID- 11934218 TI - Demonstration and quantification of "hyperchromic" erythrocytes by haematological analysers. Application to screening for hereditary and acquired spherocytosis. AB - The double laser beam diffraction of spherized RBC used in the ADVIA 120 haematological analyser allows quantitation of cells aberrant not only by their volume but also by their haemoglobin concentration. The present investigation provides arguments for the identification of hyperchromic RBC as spherocytes, mainly the close relation between % hyperchromic cells and % lysed by the cryohaemolysis test. The percentage of hyperchromic erythrocytes may no longer be considered an instrumental artefact. Without allowing a definite diagnosis of hereditary spherocytosis, an increased percentage of hyperchromic cells indicates the degree of spherocytosis, making it an excellent automated and cost-free screening parameter for inherited and acquired corpuscular haemolysis. PMID- 11934219 TI - Measurement of lipoprotein subclass profiles by nuclear magnetic resonance spectroscopy. PMID- 11934220 TI - Primary aldosteronism. AB - Primary aldosteronism is a potentially curable form of hypertension. Recent studies using the plasma aldosterone to plasma renin activity ratio as screening test in hypertensive populations have demonstrated a high prevalence of primary aldosteronism close to 10%. This frequency is clearly higher than the classically described when hypokalemia is used as the screening method. The most common subtypes of primary aldosteronism are idiopathic aldosteronism and aldosterone producing adenoma. Other causes are glucocorticoid-remediable aldosteronism, unilateral or primary adrenal hyperplasia and adrenal carcinoma. The diagnosis of primary aldosteronism is advocated to confirm the autonomy of aldosterone secretion from the renin-angiotensin system and to differentiate the clinical subtypes of the disease. This article reviews the new data about prevalence, diagnosis criteria and describes the clinical, biochemical and genetic characteristics of the different subtypes of the disease. We also discuss the treatment, and the differential diagnosis with other hyper-mineralocorticoid states. PMID- 11934221 TI - Sensitive calcitonin measurement by two-site immunometric assays: implications for calcitonin screening in nodular thyroid disease. AB - The aim of this study was to investigate the impact of analytical aspects on the clinical usefulness of calcitonin (CT) measurement. In a retrospective analysis, CT levels measured by a polyclonal immunometric assay (Scantibodies Laboratory, CA, USA) were evaluated in various clinical situations. CT in newly diagnosed medullary thyroid cancer (MTC) (n = 20) ranged from 15.5-87130 pg/ml (median 661 pg/ml). Levels >10 pg/ml were seen in 7.3% of 314 patients with benign nodules, 48.9% of 45 patients with impaired kidney function, 97.7% of 87 patients on hemodialysis, 30.2% of 43 patients after renal transplantation, and in 71.0% of 31 patients with critical illnesses. Subgroups of patients were reevaluated by two monoclonal immunometric assays specific for mature CT. CT levels measured by the monoclonal immunometric assays were highly correlated to the polyclonal assay results in MTC patients, but were significantly different with a lower incidence of elevated levels in patients with renal disease and critical illnesses. In conclusion, highly sensitive assays with cut-off values of 10 pg/ml or below are mandatory for CT screening in nodular thyroid disease. The specificity of CT measurement in patients with renal disease and critical illnesses is higher with monoclonal assays specific for monomeric CT. These methodological aspects have to be regarded if CT measurement is used for decision making in nodular thyroid disease. PMID- 11934222 TI - Humoral immune response to parvovirus B19 and serological diagnosis of B19 infection. PMID- 11934223 TI - HAMA-ELISA medac: a new assay for the quantitation of human anti-mouse antibodies. PMID- 11934224 TI - Evaluation of the chemiluminescence immunoassays for the measurement of troponin I, myoglobin and CK-MB using the IMMULITE system in comparison to other measuring systems. AB - We evaluated the chemiluminescence immunoassays for the detection of the cardiac markers troponin I, myoglobin and CK-MB on the IMMULITE System (Diagnostic Products Corporation) in comparison to the same analytes of other companies. The IMMULITE assays are two-site solid phase immunometric assays using a murine monoclonal capture antibody on the solid phase and a polyclonal antibody conjugated with alkaline phosphatase (except CK-MB monoclonal, murine) for detection. Precision was investigated using serum pools with a low, a cutoff and a high concentration of the respective analyte. The results were satisfactory with an intra-assay precision coefficient of variation, CV of 1.7% - 3.2% for troponin I, 2.6% - 5.1% for myoglobin, 2.7% - 5.3% for CK-MB and an interassay precision of 5.1% - 6.9% for troponin I, 5.7% - 7.3% for myoglobin and 3.8% - 8.4% for CK-MB. In linearity studies with various dilution steps, a mean value of 105% was found for troponin I, 103% for myoglobin and 117% for CK-MB. The average recovery was 85% for troponin I, 100% for myoglobin and 95% for CK-MB. The clinical validity of the assays in the diagnosis and therapy of myocardial infarction was investigated in 120 patients who were sent to the hospital with suspected myocardial infarction. Four hours after admission all patients with clinically verified myocardial infarction showed troponin I and troponin T values above the cutoff value. A maximum rate of 32% of the patients (IMMULITE Troponin I) with an instable angina pectoris showed troponin values above the cutoff for myocardial infarction (1.0 microg/L), 4 hours after admission. A cutoff-reduction to 0.2 pg/L for troponin I increased the number of patients to 45%. The negative predictive value was constantly 67%. The results obtained by IMMULITE assays were compared to the Elecsys cardiac assays (Roche Diagnostics) and the AxSYM-cardiac assays (Abbott Diagnostics). The highest correlation (r = 0.99) was found for IMMULITE Troponin I (DPC) and Troponin I (Abbott). The Abbott-Troponin I showed the highest diagnostic sensitivity within 4 hours after admission. All compared methods showed a similar diagnostic sensitivity (close to 100%) > 4 hours after admission. For all investigated methods the percentage of discrepant results decreased to a minimum 4 hours after admission. PMID- 11934225 TI - Chemistry, manufacturing, and controls information in NDAs and ANDAs, supplements, annual reports, and other regulatory filings. AB - Advice to the pharmaceutical industry regarding the chemistry, manufacturing, and controls and microbiology (sterility assurance) information to be included in regulatory submissions to the Center for Drug Evaluation and Research (CDER) can be found in the pertinent statutes, regulations, and guidances. The primary statute is the Federal Food, Drug and Cosmetic Act (the Act); applicable regulations appear in 21 CFR 312 and 314. Neither the Act nor the regulations provide sufficient detail on the information that should be included in these submissions. Over the past 14 years CDER has issued a series of guidelines and guidances that provide specific detail related to the recommended filing mechanisms and information that CDER expects applicants to provide. Some of these guidances are applicable to original submissions and some are applicable to post approval changes. This article will provide an overview of The Act, the pertinent regulations, and the pre- and post-approval guidances. PMID- 11934226 TI - Determination of in vivo bioequivalence. AB - The May and June 2001 issues of Pharmaceutical Research contained three articles related to the determination of in vivo Bioequivalence. The articles discussed: (a) the bioequivalence of highly variable drugs, (b) novel metrics for direct comparison of bioequivalence study plasma curves, and (c) the role of a microemulsion vehicle on cutaneous bioequivalence. An analysis of the relationship and potential impact of these articles on their respective areas of bioequivalence will be addressed in this commentary. PMID- 11934227 TI - Targeting vaccines to dendritic cells. AB - Dendritic cells (DC) are specialized antigen presenting cells (APC) with a remarkable ability to take up antigens and stimulate major histocompatibility complex (MHC)-restricted specific immune responses. Recent discoveries have shown that their role in initiating primary immune responses seems to be far superior to that of B-cells and macrophages. DC are localized at strategic places in the body at sites used by pathogens to enter the organism, and are thereby in an optimal position to capture antigens. In general, vaccination strategies try to mimic the invasiveness of the pathogens. DC are considered to play a central role for the provocation of primary immune responses by vaccination. A rational way of improving the potency and safety of new and already existing vaccines could therefore be to direct vaccines specifically to DC. There is a need for developing multifunctional vaccine drug delivery systems (DDS) with adjuvant effect that target DC directly and induce optimal immune responses. This paper will review the current knowledge of DC physiology as well as the progress in the field of novel vaccination strategies that directly or indirectly aim at targeting DC. PMID- 11934228 TI - Fundamental effects of particle morphology on lung delivery: predictions of Stokes' law and the particular relevance to dry powder inhaler formulation and development. AB - Key factors that contribute to the aerodynamic properties of aerosol particles are found in Stokes' law. These factors may be monitored or controlled to optimize drug delivery to the lungs. Predictions of the aerodynamic behavior of therapeutic aerosols can be derived in terms of the physical implications of particle slip, shape and density. The manner in which each of these properties have been used or studied by pharmaceutical scientists to improve lung delivery of drugs is readily understood in the context of aerosol physics. Additional improvement upon current aerosol delivery of particulates may be predicted by further theoretical scrutiny. PMID- 11934230 TI - Pharmaceutical evaluation of gas-filled microparticles as gene delivery system. AB - PURPOSE: To produce and characterize a nonviral ultrasound-controlled release system of plasmid DNA (pDNA) encapsulated in gas-filled poly(D,L-lactide-co glycolide) microparticles (PLGA-MPs). METHODS: Different cationic polymers were used to form pDNA/polymer complexes to enhance the stability of pDNA during microparticle preparation. The physico-acoustical properties of the microparticles, particle size, pDNA integrity, encapsulation efficiency and pDNA release behavior were studied in vitro. RESULTS: The microparticles had an average particle size of around 5 microm. More than 50% of all microparticles contained a gas core, and when exposed to pulsed ultrasound as used for color Doppler imaging create a signal that yields typical color patterns (stimulated acoustic emission) as a result of the ultrasound-induced destruction of the microparticles. Thirty percent of the pDNA used was successfully encapsulated and approximately 10% of the encapsulated pDNA was released by ultrasound within 10 min. CONCLUSIONS: Plasmid DNA can be encapsulated in biodegradable gas-filled PLGA-MPs without hints for a structural disintegration. A pDNA release by ultrasound-induced microparticle-destruction could be shown in vitro. PMID- 11934229 TI - Prevention of autoimmune insulitis by delivery of interleukin-4 plasmid using a soluble and biodegradable polymeric carrier. AB - PURPOSE: We delivered interleukin-4 (IL-4) plasmid (pCAGGS-IL-4) using the biodegradable polymer, poly[alpha-(4-aminobutyl)-L-glycolic acid] (PAGA), to prevent autoimmune insulitis in NOD mice. METHODS: The pCAGGS-IL-4/PAGA complex was transfected to 293T cells. The expression level of IL-4 was measured by ELISA. The pCAGGS IL-4/PAGA complex was injected once to NOD mice intravenously at the age of 4 weeks. RT-PCR was performed to evaluate the level of the IL-4 mRNA in the liver. At 6 weeks after the injection, the grade of insulitis of the mice was evaluated by double blind methods. RESULTS: In vitro transfecton assays showed that PAGA enhanced the expression of IL-4 in 293T cells. RT-PCR of the liver showed that IL-4 was expressed highest in the complex injected group. In the plasmid/PAGA complex injected group, the prevalence of severe insulitis in NOD mice was markedly improved, suggesting that PAGA enhanced the delivery of IL 4 plasmid. CONCLUSION: The pCAGGS-IL-4/PAGA complex is an effective system to prevent autoimmune insulitis in NOD mice and applicable for the prevention of autoimmune diabetes. PMID- 11934231 TI - In vivo lung deposition of hollow porous particles from a pressurized metered dose inhaler. AB - PURPOSE: PulmoSphere particles are specifically engineered for delivery by the pulmonary route with a hollow and porous morphology, physical diameters < 5 microm, and low tap densities (circa 0.1 g x cm(-3)). Deposition of PulmoSphere particles in the human respiratory tract delivered by pressurized metered dose inhaler (pMDI) was compared with deposition of a conventional micronized drug pMDI formulation. METHODS: Nine healthy nonsmoking subjects (5 male, 4 female) completed a two-way crossover gamma scintigraphic study, assessing the lung and oropharyngeal depositions of albuterol sulfate, formulated as 99mTc-radiolabeled PulmoSphere particles or micronized particles (Ventolin Evohaler, GlaxoSmithKline, Ltd.) suspended in HFA-134a propellant. RESULTS: Mean (standard deviation) lung deposition, (% ex-valve dose) was doubled for the PulmoSphere formulation compared with Evohaler pMDI (28.5 (11.3) % vs. 14.5 (8.1) %, P < 0.01), whereas oropharyngeal deposition was reduced (42.6 (9.0) % vs. 72.0 (8.0) %, P < 0.01). Both PulmoSphere and Evohaler pMDIs gave uniform deposition patterns within the lungs. CONCLUSIONS: These data provided "proof of concept" in vivo for the PulmoSphere technology as a method of improving targeting of drugs to the lower respiratory tract from pMDIs, and suggested that the PulmoSphere technology may also be suitable for the delivery of systemically acting molecules absorbed via the lung. PMID- 11934232 TI - Use of the post-insertion technique to insert peptide ligands into pre-formed stealth liposomes with retention of binding activity and cytotoxicity. AB - PURPOSE: Simple methods for the large-scale manufacture of ligand-targeted liposomes will be needed if clinical trials are to proceed. We tested a recently developed technology for inserting peptide ligands into preformed Stealth liposomes. Antagonist G-targeted liposomes (PLG) were prepared and loaded with doxorubicin and their cellular association and cytotoxicity were evaluated using the human small cell lung cancer H69 cell line. METHODS: The hexapeptide antagonist G was covalently coupled via a thioether bond to the terminus of polyethylene glycol (PEG) in micelles formed from maleimide-derivatized poly(ethylene glycol) (Mr 2000) distearoylphosphatidylethanolamine followed by transfer into preformed liposomes during a one-step incubation. For cellular association, we used radiolabeled liposomes. Cytotoxicity was evaluated using the MTT in vitro proliferation assay. RESULTS: The postinsertion approach to the formation of peptide-targeted liposomes led to the production of PLG bearing a maximum of approximately 0.3 microg antagonist G/micromol phospholipid. These liposomes had increased cellular association to H69 cells relative to nontargeted liposomes and, when loaded with doxorubicin, they resulted in similar levels of cytotoxicity to those obtained by conventional coupling techniques. CONCLUSIONS: The postinsertion technique is a simple, effective means for the production of biologically active peptide-targeted liposomes. PMID- 11934233 TI - Comparison of dermatopharmacokinetic vs. clinicial efficacy methods for bioequivalence assessment of miconazole nitrate vaginal cream, 2% in humans. AB - PURPOSE: To compare the dermatopharmacokinetic vs. clinical trial methods for bioequivalence assessment of two miconazole nitrate vaginal cream, 2% products. METHODS: The dermatopharmacokinetic method determined the bioequivalence of two products simultaneously in 24 healthy subjects, as a function of Cmax and AUC(0 1) parameters using miconazole nitrate content in harvested volar forearm stratum corneum. The clinical trial method determined bioequivalence as a function of clinical, mycological culture and therapeutic cure(s) after 7 days of product use and 30 days after therapy cessation in 106 female subjects with positive signs and symptoms of vaginitis, KOH vaginal smears and Candida cultures, randomly assigned to test or reference product. RESULTS: The dermatopharmacokinetic method demonstrated that the two products were not bioequivalent, while the clinical trial method concluded bioequivalence. CONCLUSION: The dermatopharmacokinetic method allowed simultaneous evaluation of both products in the same subject, within the same study period, and was more sensitive and discriminating in the assessment of bioequivalence between the two miconazole nitrate vaginal cream, 2% products than the clinical trial method. PMID- 11934235 TI - Synthesis and aromatase inhibitory activity of flavanones. AB - PURPOSE: Aromatase inhibitors are known to prevent the conversion of androgens to estrogens and play a significant role in the treatment of estrogen dependent diseases such as breast cancer. Some flavonoids have been reported as potent aromatase inhibitors; therefore, in an effort to develop novel anti breast cancer agents, B ring substituted flavanones with a 7-methoxy group on A ring were synthesized and tested to assess their ability to inhibit aromatase activity and to determine the optimal B ring substitution pattern. METHODS: A series of flavanones was prepared by cyclisation of 2'-hydroxychalcones previously obtained by Claisen-Schmidt condensation and the aromatase inhibitory activity of these compounds was investigated using human placental microsomes and radiolabeled [1,2,6,7-(3)H]-androstenedione as substrate. RESULTS: Almost all flavanones exhibited inhibitory effect on the aromatase activity but their potency was dependent on their B ring substitution pattern. Hydroxylation at position 3' and/or 4' enhanced the anti-aromatase activity; thus, 3',4'-dihydroxy-7 methoxyflavanone was found to be twice more potent than aminoglutethimide, the first aromatase inhibitor clinically used. CONCLUSIONS: These results indicated that these flavanones could be considered as potential anti breast cancer agents through the inhibition of aromatase activity and allowed us to select some of these compounds as skeleton for the development of flavonoid structurally-related aromatase inhibitors. PMID- 11934234 TI - A biodegradable injectable implant sustains systemic and ocular delivery of an aldose reductase inhibitor and ameliorates biochemical changes in a galactose-fed rat model for diabetic complications. AB - PURPOSE: To fabricate and characterize in vitro and in vivo performance of a sustained release biodegradable implant for N-4-(benzoylaminophenylsulfonyl glycine) (BAPSG), a novel aldose reductase inhibitor. METHODS: The ability of BAPSG to inhibit aldose reductase activity and glucose-induced vascular endothelial growth factor (VEGF) expression was assessed in a retinal pigment epithelial cell line (ARPE-19). A poly (DL-lactic-co-glycolic acid) implant containing 50% w/w BAPSG was fabricated and characterized for drug loading, in vitro drug release, and the thermal behavior of the drug and the polymer. Implants were injected subcutaneously into a galactose-fed diabetic rat model and cataract scores, plasma and tissue drug levels, galactitol levels in the lens and the retina, glutathione levels in the plasma, lens, cornea and retina and VEGF expression in the retina were determined on or until 18 days. RESULTS: BAPSG inhibited aldose reductase activity and reduced VEGF expression in ARPE-19 cells. Implants (1 x 4 mm), with a loading efficiency of 106 +/- 7% for BAPSG, were fabricated. Upon implant fabrication, while the glass transition temperature of the polymer decreased, the melting point of the drug was not affected. In vivo drug release correlated well with in vitro release, with approximately 44% drug release occurring in vivo by the end of 18 days. The implant reduced galactitol accumulation, glutathione depletion, cataract scores, and VEGF expression in galactose-fed rats. CONCLUSIONS: An injectable biodegradable implant of BAPSG sustained drug release in vitro and in vivo, and reduced galactitol accumulation, glutathione depletion, cataract scores, and VEGF expression in galactose-fed rats. PMID- 11934236 TI - Determining dermal absorption parameters in vivo from tape strip data. AB - PURPOSE: Tape stripping the outermost skin layer, the stratum corneum (sc), is a popular method for assessing the rate and extent of dermal absorption in vivo. Results from tape strip (TS) experiments can be affected significantly by chemical diffusion into the sc during the time required to apply and remove all of the TSs, tTS. Here, we examine the effects of this problem on the interpretation of TS experimental results. METHODS: Dermal absorption of 4 cyanophenol (4CP) in humans was studied using TS experiments to assess conditions in which diffusion alters TS results. Mathematical models were developed to assess the effects of diffusion on parameter estimation. RESULTS: For an experiment with tTS > tlag (i.e., the lag time for a chemical to cross the sc), the permeability coefficient for 4CP, P(sc,v), calculated including tTS, was consistent with values from the literature (i.e., 0.0019 cm/h). When diffusion during stripping was not included in the model, P(sc,v) was 70% smaller. CONCLUSIONS: Calculations show that chemical concentrations in TSs can be affected by diffusion during tape stripping, but if tTS < 0.2 tlag and the exposure time is > 0.3 tlag, TS concentrations are not significantly affected by tTS. PMID- 11934237 TI - Sodium dependence of nitrofurantoin active transport across mammary epithelia and effects of dipyridamole, nucleosides, and nucleobases. AB - PURPOSE: The sodium dependence and effects of nucleoside and nucleobase transport inhibitors were determined to ascertain the role of sodium dependent nucleoside or nucleobase transporters in nitrofurantoin active transport across mammary epithelia. METHODS: Five lactating female rats received steady-state intravenous infusions of nitrofurantoin with and without the broad-based inhibitor dipyridamole. In the CIT3 murine model of lactation, 14C-nitrofurantoin basolateral to apical permeability was examined in the presence of varying sodium concentrations, purine and pyrimidine nucleosides and nucleobases, and dipyridamole. RESULTS: Dipyridamole effectively inhibited 14C-nitrofurantoin flux across CIT3 cells, with Ki = 0.78 microM (95% C.I. = 0.11 to 5.3 microM) and significantly decreased the milk-to-serum ratio of nitrofurantoin from 29.2 +/- 5.0 to 11.0 +/- 6.3 without changing systemic clearance. Nitrofurantoin active transport was significantly inhibited by complete sodium replacement. Adenosine and guanosine significantly inhibited nitrofurantoin permeability (54.5 +/- 2.6 (microl/hr)/cm2 and 50.7 +/- 0.6 (microl/hr)/cm2, respectively, vs. control 90.5 +/- 4.6 (microl/hr)/cm2) but uridine, thymidine, and the nucleobases had no effect. CONCLUSIONS: Nitrofurantoin active transport was sodium dependent and inhibited by dipyridamole, adenosine, and guanosine, but known sodium dependent nucleoside or nucleobase transporters were not involved. PMID- 11934238 TI - Understanding and predicting drug delivery from hydrophilic matrix tablets using the "sequential layer" model. AB - PURPOSE: The objectives of this work were (i) to study and understand the physicochemical phenomena which are involved in the swelling and drug release from hydrophilic matrix tablets using the "sequential layer" model, and (ii) to predict the effect of the initial radius height and size of the tablets on the resulting drug release profiles. METHODS: Tablets were prepared by direct compression, using hydroxypropyl methylcellulose (HPMC) grades with different average molecular weights as matrix-forming polymers. The in vitro release of chlorpheniramine maleate, propranolol HCl, acetaminophen, theophylline and diclofenac sodium was studied in phosphate buffer (pH 7.4) and 0.1 M HCl, respectively. The initial drug loading varied from 1 to 70%, while the radius and height of the tablets varied from 1 to 8 mm. RESULTS: The "sequential layer" model considers water and drug diffusion with non-constant diffusivities and moving boundary conditions, non-homogeneous polymer swelling, drug dissolution, and polymer dissolution. We showed that this model was able to predict the resulting drug release kinetics accurately in all cases. CONCLUSIONS: The "sequential layer" model can be used to elucidate the swelling and drug release behavior from hydrophilic matrix tablets and to simulate the effect of the device geometry on the drug release patterns. Hence, it can facilitate the development of new pharmaceutical products. PMID- 11934239 TI - Prediction of poly(ethylene) glycol-drug eutectic compositions using an index based on the van't Hoff equation. AB - PURPOSE: To define an index based on the van't Hoff equation that can be used as a screening tool for predicting poly(ethylene) glycol (PEG)-drug eutectic composition. METHODS: Phase diagrams of PEG with ritonavir, ibuprofen, fenofibrate. naproxen, and griseofulvin were constructed using differential scanning calorimetry, hot stage microscopy and powder X-ray diftractometry. Previously reported phase diagrams were also used to test the predictive capability of the index. RESULTS: This work shows that a modified van't Hoff equation can be used to model the drug liquidus line of these phase diagrams. The slope of the liquidus line depends on the melting point (T(f)d) and heat of fusion (deltaH(f)d) of the drug and describes the initial rate at which the eutectic or monotectic point is approached. Based on this finding, a dimensionless index Ic was defined. The index can be calculated from the melting points of the pure components and heat of fusion of the drug. In addition to the compounds listed above, the index was found to predict the eutectic composition for flurbiprofen, temazepam and indomethacin. These compounds range over 150 degrees C in T(f)d, and from 25-65 kJ/mole in deltaH(f)d. CONCLUSION: Using Ic the approximate eutectic composition for eight different compounds was predicted. The index provides a useful screening tool for assessing the maximum drug loading in a drug-polymer eutectic/monotectic formulation. PMID- 11934240 TI - Adhesion of powders for inhalation: an evaluation of drug detachment from surfaces following deposition from aerosol streams. AB - PURPOSE: To evaluate micronized powder retention and detachment from inhaler surfaces following reproducible deposition by impaction, coupled with centrifugal particle detachment (CPD). METHODS: Micronized albuterol sulfate (AS) and beclomethasone dipropionate (BDP) were aerosolized as dry powders and deposited by cascade impaction onto different contact surfaces. Drug detachment from the surfaces was characterized using CPD, coupled with HPLC assay and scanning electron microscopy. RESULTS: Drugs which accumulated as aggregates on model surfaces detached with distinctive profiles for % remaining vs. applied centrifugal force; each profile showed reproducible values for the minimum force required to initiate drug detachment, Fyield. While differences occurred in the observed detachment profiles for different drugs and contact surfaces (polyacetal vs. aluminum), the deposited drug particle size had the most significant effect on these profiles, e.g., Fyield for AS (2.1-3.3 microm) was 383 +/- 12.7 microN compared with 18 +/- 13.8 microN for AS (4.7-5.8 microm). CONCLUSIONS: A technique was developed which enabled the experimental review, and subsequent data analysis, of the adhesive properties between different DPI construction materials and drug substances deposited from aerosol clouds. The technique appears to be of greater relevance to inhaler design decisions than earlier studies in the literature claiming to show differences in the adhesion of single drug particles to surfaces. PMID- 11934241 TI - Neotame anhydrate polymorphs I: preparation and characterization. AB - PURPOSE: To prepare, characterize, and compare polymorphs of neotame anhydrate. METHODS: Neotame anhydrate polymorphs were prepared from amorphous or crystalline anhydrate by crystallization or suspension in various organic solvents, or by dehydration of neotame monohydrate. The following techniques were used for characterization: differential scanning calorimetry, thermogravimetry, hot-stage microscopy, powder X-ray diffractometry (PXRD), 13C solid-state nuclear magnetic resonance (SSNMR) spectroscopy, Fourier transform infrared (FTIR) spectroscopy, scanning electron microscopy, dynamic water vapor sorption/desorption, and density measurements. RESULTS: Seven polymorphs (Forms A-G) of neotame anhydrate were prepared and show different thermal properties and PXRD patterns. Two enantiotropically related pairs were identified: B and C; E and A. 13C SSNMR and FTIR spectroscopy clearly distinguish between Forms A, D, F, and G, which show similar needle-shaped morphology but distinct differences in dynamic water vapor sorption/desorption and density. The 13C SSNMR chemical shifts suggest conformational polymorphism. The stability in the presence of water vapor follows the rank order, G > A > D approximately = F, which resembles the rank orders of the molar volume and of the polarity of the solvents from which they crystallized. CONCLUSIONS: The neotame anhydrate polymorphs appear to show different molecular conformations. The less dense polymorphic structures crystallize from solvents of greater polarity and sorb water vapor less rapidly and less completely. Two enantiotropic pairs were discerned. PMID- 11934243 TI - Forecasting the oral absorption behavior of poorly soluble weak bases using solubility and dissolution studies in biorelevant media. PMID- 11934242 TI - Development of a remote electrode system for monitoring the water content of materials inside a glass vial. AB - PURPOSE: This article explores the use of a remote electrode dielectric measurement system to monitor the water content of hydrated ovalbumin inside a glass vial. METHODS: The intrinsic dielectric properties of hydrated ovalbumin were characterized first using conventional parallel plate electrodes. The second stage was to simulate a remote electrode measurement by placing nonconductive, nondispersive polyethylene films between the sample and electrodes. Finally, a study on the dielectric measurement of ovalbumin contained in a 10 ml glass vial was undertaken with the electrodes external to the glass vial. RESULTS: The dielectric behavior of hydrated ovalbumin was characterized by charge transfer (i.e., protons) in the hydrogen bonded network of water molecules in the bulk sample. The mechanism was identified as an anomalous low-frequency dispersion and a dielectric loss peak (epsilon3). The dielectric relaxation time, tau3, of the epsilon3 dispersion was especially sensitive to water content. Moreover, a good correlation (R2 = 93%) was observed between relaxation times tau3 obtained from measurements using conventional parallel plate electrodes and the remote electrode system. CONCLUSIONS: Dielectric measurements using remote electrodes attached to a glass vial are therefore applicable for the in situ measurement of water content in materials. The application of this technology to the determination of the lyophilization end point is suggested. PMID- 11934244 TI - A new method to determine the distribution of a fluorophore in scalp skin with focus on hair follicles. PMID- 11934245 TI - Effects of elements in human blood pressure control. AB - This review enumerates and discusses the elements involved in the control of human blood pressure via a historical evolutionary form. The older and most recent element literature presentations were researched using MEDLINE and a manual review of documents cited. Independent data extraction and cross referencing was performed. Of the 28 known elements that can influence blood pressure, 15 were found to be involved in human blood pressure regulation. The elements were divided into four groups: electrolyte, composed of sodium, potassium, calcium, and magnesium; metal, which included zinc, copper, and iron; toxic, made up of lead, mercury, cadmium, barium, thallium, arsenic; miscellaneous (lithium and selenium). Evolutionary historical data, possible mechanisms of actions, and interactions between elements that have been shown to influence blood pressure are discussed. Controversy exists over the therapeutic use of elements to alter blood pressure but is absent in the case of the toxic group where preventive control is a proven public health matter. The significance of these 15 elements in the regulation of human blood pressure has been established and ongoing studies will continue to reinforce their influence and importance. PMID- 11934246 TI - Phosphate deposition capacity of athletes during hypokinesia, phosphate loading, and ambulation. AB - Hypokinesia (diminished movement) induces significant phosphate (P) excretion; however, little is known about the P deposition ability of the body during hypokinesia (HK). Using P loads, the aim of this study was to establish the deposition ability of the body to retain P during prolonged HK. Studies were done during a 30-d period of pre-HK and a 364-d period of HK. Forty male trained athletes aged 24.7 +/- 8.0 yr were chosen as subjects. They were equally divided into four groups: unloaded ambulatory control subjects (UACS), unloaded hypokinetic subjects (UHKS), loaded ambulatory control subjects (LACS), and loaded hypokinetic subjects (LHKS). All hypokinetic subjects were limited to an average walking distance of 0.7 km/d. Loading tests with 85.0 mg of calcium phosphate/kg body weight were performed on the LACS and LHKS. Fecal P loss, urinary calcium (Ca) and P loss, serum P, Ca, and the ionized calcium (CaI) levels increased significantly (p < or = 0.05) in the LHKS and UHKS groups when compared with the LACS and UACS groups, respectively. Serum intact parathyroid hormone (iPTH) and the 1,25-dihydroxyvitamin D3 [1,25-(OH)2 D3] levels decreased significantly (p < or = 0.05) in the LHKS and UHKS groups when compared with the LACS and UACS groups, respectively. After the P load, significant (p < or = 0.05) differences were observed between LHKS and UHKS groups regarding serum, urinary, and fecal P changes. Thus, the deposition capacity of P decreased significantly (p < or = 0.05) more in the LHKS group than in the UHKS group. The deposition of P, fecal P, urinary P and Ca, serum CaI, P, Ca, 1,25-(OH)2 D3, and iPTH changed insignificantly (p > 0.05) in control groups when compared with their baseline values. It was shown that after the P load, significant differences were observed between the loaded and unloaded hypokinetic subjects regarding serum, urinary, and fecal P values and P retention. The oral P load intensified P loss from the body. It was concluded that the higher the P intake increased the greater P loss and the lower P deposition and thus the less likely it is for the P load to benefit hypokinetic subjects. PMID- 11934247 TI - Influence of age, sex, and sexual activity on trace element levels and antioxidant enzyme activities in field mice (Apodemus sylyaticus and Mus spretus). AB - The influence of age, gender and sexual activity on both hepatic levels of some trace elements (Fe, Cu, Zn, Mn, Se) and the activities of glutathione-S transferase (GST) and superoxide dismutase (SOD) was investigated in Wood mice (Apodemus sylvaticus) and Algerian mice (Mus spretus). Animals were taken from a riverside community of an unpolluted area of central Portugal. Adult A. sylvaticus presented the highest hepatic mean concentrations of Cu and Mn, whereas adult M. spretus had the highest Fe concentration in the liver. Moreover, an influence of age on the contents of Fe, Zn, and Mn has been observed in A. sylvaticus, whereas in M. spretus an influence of gender and sexual activity was only detected on Zn levels. In contrast, enzyme activities were not influenced by the studied variables, despite a tendency for an increase in SOD activity in sexually active M. spretus. GST activity was species dependent, whereas SOD activity was similar between species. These findings were analyzed regarding the relationship of both essential trace elements and the two antioxidant enzymes with physiological and metabolic pathways related to life cycles in the two species of mice. Results enhanced the understanding of A. sylvaticus and M. spretus as biological models, allowing their future use as bioindicators of environmental toxicity. PMID- 11934249 TI - Study of trace element correlations with drought tolerance in different sorghum genotypes using energy-dispersive X-ray fluorescence technique. AB - Drought-tolerant and drought-susceptible genotypes of sorghum (Sorghum bicolor L. Monech) were analyzed by the energy-dispersive X-ray fluorescence (EDXRF) technique to study the correlation of trace elements with drought-tolerance capacities. Samples prepared from mature seeds, young seedlings, and old plants were analyzed using a 109Cd radioisotope source and a Si(Li) semiconductor detector of resolution 170 eV for 5.9-keV MnKalpha X-rays. Elements such as K, Fe, Cu, Zn, Rb and Sr and Y were found to be present in varying concentrations in different samples. The trace element profile studied in the seeds of 11 genotypes and in seedlings (young and old) of 4 sorghum genotypes showed considerable variation. The genotype Arfa Gadamak (AG) showed a distinct presence of a high level of Zn in its young seedling. It was observed that in most of the genotypes (seeds), K and Fe concentrations were more in the tolerant genotype as compared to the susceptible type. The concentration of Fe decreased with maturity in the tolerant group and it increased with maturity in the susceptible group. PMID- 11934248 TI - Reactive oxygen species from mitochondria mediate SW480 cells apoptosis induced by Na2SeO3. AB - A number of selenium compounds have been found to inhibit tumorigenesis in a variety of animal and cell models. In order to explore the molecular mechanism involved in the anticarcinogenesis activity of selenium, we examined the effects of sodium selenite on cell viabilty, generation of reactive oxygen species (ROS), and mitochondrial transmembrane potential (delta(psi)m) in human colonic carcinoma cells SW480. The result from MTT test showed that sodium selenite reduced cell viability. Morophologic and flow cytometric results indicated that Na2SeO3 induced the apoptosis of SW480 cells. Na2SeO3 increased the generation of intracellular ROS, whereas BAPTA-AM, rotenone, and NaCN completely inhibited the increase of ROS induced by Na2SeO3. Na2SeO3 also caused the disruption of delta(psi)m. The intracellular ROS increase and apoptosis induced by Na2SeO3 were significantly decreased by superoxide dismutase (SOD), catalase. These data suggest that the ROS mediate apoptosis induced by Na2SeO3 and mitochondria may be a major source of Na2SeO3-induced ROS. PMID- 11934250 TI - Hair zinc and copper in chronic alcoholics. AB - Chronic alcoholics frequently show associated malnutrition. Both ethanol and malnutrition exert profound changes on zinc and copper metabolism. In this study, we found higher hair zinc and copper values in 43 male alcoholics than in 39 controls. Hair copper was significantly related to the amount of ethanol consumed, whereas hair zinc was higher in consumers of distilled beverages. No relation was observed between hair zinc and copper and nutritional status, kind of diet consumed, style of life, and liver cirrhosis. Consequently, hair zinc and copper levels are related only with alcohol intake. PMID- 11934251 TI - Selenium levels in human plasma and hair in northern Poland. AB - The aim of this study was to (1) estimate the concentration of selenium in the plasma of 146 residents (65 men and 81 women) and in the hair of 34 persons from the Gdansk region in northern Poland, aged 19-70 and (2) compare the obtained results with data corresponding to healthy populations living in different European countries. Selenium in plasma was determined by atomic absorption spectrometry using the hydride generation method. The mean selenium concentration in plasma of the investigated persons was 73.3 +/- 14.1 microg/L, 76.7 +/- 13.2 microg/L in men, and 70.4 +/- 14.7 microg/L in women. No age - dependent differences in plasma selenium were found in the investigated population. In 20% of the investigated persons, the selenium level in plasma was lower than 60 microg/L. The mean selenium concentration in hair was 0.30 +/- 0.11 microg/g. A positive, statistically significant correlation between selenium concentrations in the plasma and hair of the investigated persons was found. The obtained results indicate that the selenium level in significant part of this population is suboptimal and should be elevated by supplementation with this element. PMID- 11934252 TI - Oxygen as a regulator of cellular phenotypes in pregnancy and cancer. AB - Cellular phenotype is determined by genetic and microenvironmental factors. There is evidence that tissue oxygenation status is one of the microenvironmental factors regulating cellular behaviour. Both normal and pathological processes such as blastocyst implantation in the uterus, placentation, and rapidly growing tumours occur under conditions characterized by relatively low oxygen levels. In this review, we address the effects of low oxygen concentrations on the phenotype of trophoblast and cancer cells. We provide evidence that oxygenation levels play an important role in the regulation of normal and pathological cellular invasiveness as it occurs during trophoblast invasion of the uterus and in tumour progression and metastasis, drug resistance in cancer, and antitumour activity of natural killer cells of the immune system. PMID- 11934253 TI - Trophectoderm development and function: the roles of Na+/K(+)-ATPase subunit isoforms. AB - Preimplantation development is a period of cell division, cell shape change, and cell differentiation leading to the formation of an epithelium, the trophectoderm. The trophectoderm is the part of the conceptus that initiates uterine contact and, after transformation to become the trophoblast, uterine invasion. Thus, trophectoderm development during preimplantation stages is a necessary antecedent to the events of implantation. The preimplantation trophectoderm is a transporting epithelium with distinct apical and basolateral membrane domains that facilitate transepithelial Na+ and fluid transport for blastocoel formation. That transport is driven by Na+/K(+)-ATPase localized in basolateral membranes of the trophectoderm. Preimplantation embryos express multiple alpha and beta subunit isoforms of Na+/K(+)-ATPase, potentially constituting multiple isozymes, but the basolaterally located alpha1beta1, isozyme uniquely functions to drive fluid transport. They also express the gamma subunit, which is a modulator of Na+/K(+)-ATPase activity. In the mouse, two splice variants of the gamma subunit, gammaa and gammab, are expressed in the trophectoderm. Antisense knockdown of gamma subunit accumulation caused a delay of cavitation, implying an important role in trophectoderm function. The preimplantation trophectoderm offers a unique model for understanding the roles of Na+/K(+)-ATPase subunit isoforms in transepithelial transport. PMID- 11934254 TI - Regulation of human trophoblast migration and invasiveness. AB - The human placenta is an invasive structure in which highly proliferative, migratory, and invasive extravillous trophoblast (EVT) cells migrate and invade the uterus and its vasculature. Using in vitro propagated normal first-trimester EVT cells and immortalized EVT cells, which share all of the phenotypic and functional characteristics of the normal EVT cells, it has been shown that migration/invasion of human EVT cells is stringently regulated by many growth factors, their binding proteins, extracellular matrix (ECM) components, and some adhesion molecules in an autocrine/paracrine manner at the fetal-maternal interface in human pregnancy. Transforming growth factor beta (TGF-beta), decorin (a proteoglycan in the ECM), and melanoma cell adhesion molecule (Mel-CAM) inhibit, and insulin-like growth factor II (IGF-II), IGF-binding protein 1 (IGFBP 1), and endothelin 1 (ET-1) stimulate EVT cell migration/invasion. Inhibition of EVT cell migration by TGF-beta has been suggested to be due to upregulation of integrins, which make the cells more adhesive to the ECM. Its antiinvasive action is due to an upregulation of tissue inhibitor of matrix metalloprotease 1 (TIMP 1) and plasminogen activator inhibitor (PAI-1) and a downregulation of urokinase type plasminogen activator (uPA). Molecular mechanisms of inhibition of migration/invasion of EVT cells by decorin and Mel-CAM remain to be identified. IGF-II action has been shown to be mediated by IGF type I receptors (IGF-RII) independently of IGF type I receptors (IGF-RI) and IGFBPs. This action of IGF-II appears to involve inhibitory G proteins and phosphorylation of mitogen-activated protein kinase (MAPK) (extracellular signal-regulated protein kinases 1 and 2 (ERK-1 and ERK-2)). IGFBP-1 stimulation of EVT cell migration appears to occur by binding its Arg-Gly-Asp (RGD) domain to alpha5beta1 integrin, leading to phosphorylation of focal adhesion kinase (FAK) and MAPK (ERK-1 and ERK-2). These studies may improve our understanding of diseases related to abnormal placentation, viz. hypoinvasiveness in preeclampsia and hyperinvasiveness in trophoblastic neoplasms. PMID- 11934255 TI - Role of nitric oxide in tumour progression with special reference to a murine breast cancer model. AB - Nitric oxide (NO) is a potent bioactive molecule produced in the presence of NO synthase (NOS) enzymes, which mediates numerous physiological functions under constitutive conditions. Sustained overproduction of NO (and NO-reaction products), typically under inductive conditions, can lead to cell cycle arrest and cellular apoptosis. Furthermore, carcinogenesis may result from mutational events following NO-mediated DNA damage and hindrance to DNA repair (e.g., mutation of tumour-suppressor gene p53). In a majority of human and experimental tumours, tumour-derived NO appears to stimulate tumour progression; however, for a minority of tumours, the opposite has been reported. This apparent discrepancy may be explained by differential susceptibility of tumour cells to NO-mediated cytostasis or apoptosis, and the emergence of NO-resistant and NO-dependent clones. NO-resistance may be mediated by p53 inactivation, and upregulation of cyclo-oxygenase-2 and heat shock protein 70 (HSP70). In a murine mammary tumour model, tumour-derived NO promoted tumour growth and metastasis by enhancing invasive, angiogenic, and migratory capacities of tumour cells. Invasion stimulation followed the altered balance of matrix metalloproteases and their inhibitors; migration stimulation followed activation of guanylate cyclase and MAP kinase pathways. Selective NOS inhibitors may have a therapeutic role in certain cancers. PMID- 11934256 TI - Gap junctions and tumour progression. AB - Gap junctional intercellular communication has been implicated in growth control and differentiation. The mechanisms by which connexins, the gap junction proteins, act as tumor suppressors are unclear. In this review, several different mechanisms are considered. Since transformation results in a loss of the differentiated state, one mechanism by which gap junctions may control tumour progression is to promote or enhance differentiation. Processes of differentiation and growth control are mediated at the genetic level. Thus, an alternative or complimentary mechanism of tumour suppression could involve the regulation of gene expression by connexins and gap junctional coupling. Finally, gap junction channels form a conduit between cells for the exchange of ions, second messengers, and small metabolites. It is clear that the sharing of these molecules can be rather selective and may be involved in growth control processes. In this review, examples will be discussed that provide evidence for each of these mechanisms. Taken together, these findings point to a variety of mechanims by which connexins and the gap junction channels that they form may control tumour progression. PMID- 11934257 TI - Human placental trophoblast as an in vitro model for tumor progression. AB - The human placenta is a highly invasive tumor-like structure in which a subpopulation of placental trophoblast cells known as the "extravillous trophoblast" (EVT) invades the uterine decidua and its vasculature to establish adequate fetal-maternal exchange of molecules. By utilizing in vitro-propagated short-lived EVT cell lines we found that molecular mechanisms responsible for their invasiveness are identical to those of cancer cells; however, unlike cancer cells, their proliferation, migration, and invasiveness in situ are stringently controlled by decidua-derived transforming growth factor (TGF)-beta. By SV40T antigen transfection of normal EVT cells followed by a forced crisis regimen in culture we produced an immortalized premalignant derivative that is hyperproliferative, hyperinvasive, and deficient in gap-junctional intercellular communication. Both premalignant and malignant EVT (JAR and JEG-3 choriocarcinoma) cell lines were found to be TGF-beta-resistant. Using these cell lines, we investigated genetic changes responsible for transition of the normal EVT cells to premalignant and malignant phenotype. Hyperinvasiveness in both cases resulted from a downregulation of tissue inhibitor of metalloprotease (TIMP)-1 and plasminogen activator inhibitor (PAI)-1 genes. In contrast to normal EVT cells, both cell types failed to upregulate these genes in response to TGF beta. Loss of TGF-beta response in malignant EVT cells was explained by the loss of expression of Smad3 gene. Differential mRNA display of normal and premalignant EVT cells identified up- and down-regulation of numerous known or novel genes in premalignant EVT cells, with potential oncogenic and (or) tumor-suppressor functions, e.g., loss of fibronectin and insulin-like growth factor binding protein (IGFBP-5). Premalignant EVT cells also lost IGF receptor type 2 (IGFR II). IGFBP-5 was shown to be a negative regulator of IGF-1-induced proliferation of premalignant EVT cells, so that loss of IGFBP-5 as well as IGFR-II permitted their unrestricted proliferation in an IGF-I-rich microenvironment of the fetal maternal interface. The present model may be a good prototype for identifying genetic changes underlying epithelial tumor progression. PMID- 11934258 TI - Studies on the angiotensin-converting enzyme and the kinin B2 receptor in the rabbit jugular vein: modulation of contractile response to bradykinin. AB - The rabbit jugular vein (rbJV) was used as a bioassay system to validate some early and new hypothetical interactions between the angiotensin-converting enzyme (ACE) and the B2 receptor, which may be influenced by ACE inhibitors (ACE-I). These involve the potentiation of the contractile effect of bradykinin (BK) and BK analogues, which are inactivated by ACE (e.g., [Hyp3, Tyr(Me8)]-BK (R556)), the prevention of BK-induced B2 receptor desensitisation, and the restoration of receptor sensitivity in tissues desensitised with B2 receptor agonists. Enzymatic degradation studies performed in vitro and in vivo revealed that BK and R556 are readily degraded by rabbit ACE whereas [Phe8psi(CH2-NH)Arg9]-BK (R379) is totally resistant. BK, R556, and R379 contracted endothelium-denuded veins with similar potencies (pEC50 range 8.10-8.50). Tissues pretreated with ACE-I showed an increase in pEC50 values for BK and R556 but not for R379. ACE-I (captopril, enalaprilat) were unable to prevent B2 receptor desensitisation induced by BK (1 microM). ACE-I partially restored B2 receptor-mediated contraction in tissues initially exposed to BK but not to R379. These effects were antagonised by HOE 140 (0.1 microM) but were unaffected by AcLys[Dbeta-Nal7, Ile8]-desArg9BK (R715) (1 microM) or by Losartan (1 microM). In conclusion, the potentiation of BK and its analogues relates exclusively on prevention of their metabolism, B2 receptor desensitisation is not affected by ACE-I, and restoration of tissue responsiveness to BK by ACE-I may be attributed to changes in BK concentrations in the vicinity of the B2 receptor. PMID- 11934259 TI - Glyconeogenic pathway in isolated skeletal muscles of rats. AB - Although the conversion of lactate to glycogen (glyconeogenesis) in muscle was demonstrated a long time ago, the biochemical reactions responsible for this process are still a controversial matter. In the present study, advantage was taken from the specific inhibition induced by phenylalanine on muscle pyruvate kinase (PK) to investigate the role of reverse PK activity in muscle glyconeogenesis. Addition of phenylalanine to the incubation medium of a preparation of isolated, intact skeletal muscles that maintain metabolic activity for several hours reduced by 50% the rate of incorporation of [14C]lactate or [14C]bicarbonate into muscle glycogen. Muscle extracts presented high levels of maximal activity of PK in the reverse direction, which was completely blocked in the presence of phenylalanine. In contrast, mercaptopicolinic acid, an inhibitor of phosphoenolpyruvate carboxykinase (PEPCK), did not affect the incorporation of 14C from either lactate or bicarbonate into muscle glycogen. Maximal PEPCK activity was much lower in muscle extracts than in gluconeogenic or glyceroneogenic tissues and was suppressed in the presence of mercaptopicolinic acid. The data suggest that a reversal of the metabolic flux through the reaction catalyzed by PK contributes to the accumulation of lactate-derived glycogen that occurs in skeletal muscle under certain physiological conditions. PMID- 11934260 TI - Inhibition of angiogenesis by two-chain high molecular weight kininogen (HKa) and kininogen-derived polypeptides. AB - We recently reported that the two-chain form of human high molecular weight kininogen (HKa) inhibits angiogenesis by inducing endothelial cell apoptosis (Zhang et al. 2000). This property appears to be primarily conferred by HKa domain 5 (HKa D5). In this manuscript, we further characterize the activity of these polypeptides toward proliferating endothelial cells, as well as their in vivo anti-angiogenic activity in the chick chorioallantoic membrane (CAM). We also demonstrate that short peptides derived from endothelial cell binding regions in HKa domains 3 and 5 inhibit endothelial cell proliferation and induce endothelial cell apoptosis. Like HKa and HKa D5, peptides derived from the latter domain induce endothelial cell apoptosis in a Zn(2+)-dependent manner, while those derived from domain 3 function independently of Zn2+. The implications of these findings to the regulation of angiogenesis and development of anti angiogenic therapeutics are discussed. PMID- 11934261 TI - The role of hepatocyte growth factor (scatter factor) in epithelial-mesenchymal transition and breast cancer. AB - North American women have a one in eight lifetime risk of developing breast cancer, and approximately one in three women with breast cancer will die of metastases. We, and others, have recently shown that high levels of expression of hepatocyte growth factor (HGF) and its receptor Met are associated with invasive human breast cancer and may be causally linked to metastasis. This high level of HGF and Met expression has been considered as a possible indicator of earlier recurrence and shortened survival in breast cancer patients. In contrast, HGF expression (but not Met) is strongly suppressed in normal breast epithelial cells. HGF and Met are therefore candidate targets for therapeutic intervention in the treatment of breast cancer. We have recently demonstrated that sustained activation or hyper-activation of c-Src and Stat3, which occurs in invasive breast cancer, can stimulate strong expression of HGF in carcinoma cells. In contrast, transient induction of Stat3 occurs in normal epithelium and promotes mammary tubulogenesis. We hypothesize that increased autocrine HGF-Met signaling is a critical downstream function of c-Src-Stat3 activation in mammary tumorigenesis. Future studies will identify novel Stat3 consensus sites that regulate HGF promoter activity and HGF expression preferentially in carcinoma cells and could lead to novel therapeutic drugs that specifically block HGF expression in mammary carcinoma cells, and which could be used in combined treatments to abrogate metastasis. PMID- 11934262 TI - The early life of a Schwann cell. AB - Schwann cells are the major glial population of the vertebrate peripheral nervous system. In the adult, they build a protecting sheath around neuronal processes and myelinate large-caliber axons. Already early in development, Schwann cells and neurons establish close contacts. Later development and the maintenance of peripheral nerves are crucially dependent on the controlled bi-directional dialogue between these two cell types. Several major phases can be distinguished in the life of a Schwann cell: determination, differentiation, and potentially myelination. The aim of this review is to summarize the molecular and cellular characteristics of the first steps in the life of a Schwann cell, the development from a multipotent neural crest cell to a differentiated Schwann cell. PMID- 11934264 TI - Molecular structure and alternative splicing of the human carboxypeptidase M gene. AB - Using RACE technology the 5' and 3' ends of human carboxypeptidase M (CPM) mRNA were determined and found to be divergent from the published sequence. With these results the complete structure of the human CPM gene was established based on the human genome sequence in the GenBank database. The gene was shown to contain 9 exons comprising at least 75 kb of genomic sequence. A novel first exon of 30 bp was identified and an upstream promoter sequence containing several transcription factor binding sites was found by computer analysis. Furthermore, the ATG starting codon was detected defining an open reading frame of 1329 bp that codes for a protein of 443 residues. Additionally, the polyadenylation site was discovered, determining a 3' noncoding region of 2000 nucleotides. The exon intron boundaries diverged substantially compared to those of the other basic carboxypeptidases, CPD, CPE, CPN, and AEBP1. Cloning and sequencing of RT-PCR products from different tissues revealed alternative splicing of exons 3 and 5, which results in the generation of four different mRNA isoforms. RNA extracted from tumor tissues contained more CPM mRNA than control tissue, suggesting an upregulation of CPM expression in tumors and raising the question of the role of this enzyme in cancer. PMID- 11934263 TI - TCF: Lady Justice casting the final verdict on the outcome of Wnt signalling. AB - The Wnt signalling cascade plays an important role during embryonic patterning and cell fate determination and is highly conserved throughout evolution. Factors of the TCF/LEF HMG domain family (Tcfs) are the downstream effectors of this signal transduction pathway. Upon Wnt signalling, a cascade is initiated that results in the translocation of beta-catenin to the nucleus, where it interacts with Tcf to generate a transcriptionally active complex. This bipartite transcription factor is targeted to the upstream regulatory regions of Tcf target genes. In the absence of Wnt signals, beta-catenin is degraded in the cytoplasm via the ubiquitin-proteasome pathway. Several proteins are instrumental in achieving this tight regulation of beta-catenin levels in the cell, including adenomatous polyposis coli (APC), GSK3 beta, and Axin/Conductin. Deregulation of the Wnt signalling pathway is implicated in several forms of cancer, such as colon carcinoma and melanoma. This deregulation is achieved via mutation of APC, beta-catenin or Axin, resulting in elevated beta-catenin levels and the presence of constitutively active Tcf-beta-catenin complexes in the nucleus. The accompanying inappropriate activation of target genes is considered to be a critical, early event in this carcinogenesis. In addition to regulating beta catenin levels, normal healthy cells have evolved a second level of regulation, by manipulating the activity of the Tcf proteins themselves. In the absence of Wnt signalling, Tcf complexes with several transcriptional repressor proteins ensuring active repression of Tcf target genes. In this review the dual role of Tcf proteins in the Wnt signalling cascade will be discussed. PMID- 11934265 TI - DeltaF508-CFTR causes constitutive NF-kappaB activation through an ER-overload response in cystic fibrosis lungs. AB - The clinical course of Cystic Fibrosis is characterized by recurrent pulmonary infections which ultimately lead to death by respiratory failure. The most common CF causing mutation, deltaF508-CFTR, produces an incorrectly folded protein, which accumulates within the endoplasmic reticulum. However, the molecular mechanism by which the deltaF508-CFTR protein facilitates pulmonary infection and inflammation remains unclear. Here we show that the expression of deltaF508-CFTR causes a constitutive activation of the pro-inflammatory transcription factor NF kappaB by eliciting an ER stress reaction, the ER-overload response. This endogenous NF-kappaB activation stimulates the transcription of pro-inflammatory cytokines thereby commencing an inflammatory cascade within the CF lung. PMID- 11934266 TI - Cultivation in glucose-deprived medium stimulates mitochondrial biogenesis and oxidative metabolism in HepG2 hepatoma cells. AB - In order to test the hypothesis that an imbalance between energy requirement and energy supply regulates mitochondrial genes and ultimately mitochondrial biogenesis, energy supply was challenged in HepG2 cells by withdrawal of glucose from the culture medium, making the cells exclusively dependent on mitochondrial ATP production. Such cells showed a 2-fold increase of cytochrome c oxidase activity, elevated levels of mitochondrial DNA, mitochondrial DNA encoded mRNAs and proteins, as well as the nuclear encoded mitochondrial transcription factor A. Lactate production was significantly reduced and glutamine was consumed as an alternative substrate for oxidative metabolism. Long-term adapted cells formed exclusively monolayers, while they normally grow in multilayers forming tumor spheroids. Also, long-term adapted cells proliferated significantly faster. No differences for the ATP/ADP ratio were observed, indicating that this is not the primary signal initiating the adaptative processes. These results show that mitochondrial biogenesis and oxidative metabolism are stimulated in HepG2 cells grown in the absence of fermentable glucose, probably in order to compensate for the diminished supply of glycolytic ATP. PMID- 11934267 TI - Intracellular concentration measurements in adherent cells: a comparison of import efficiencies of cell-permeable peptides. AB - A protocol was developed for performing intracellular concentration measurements in flat adherent tissue culture cells by fluorescence correlation microscopy (FCM). Determination of the number of molecules in the confocal detection volume had to account for background fluorescence caused by molecules adsorbed to the surface of the measurement chamber. Such a background signal leads to a decrease in the amplitude of the autocorrelation function, and thereby to the calculation of an erroneously high number of molecules. Because of the spatial heterogeneity of the background intensity, a method was devised by which its contribution to the total fluorescence could be determined directly from each individual autocorrelation measurement. This method was applied to a comparison of the import efficiencies of different cell-permeable peptides at nanomolar concentrations. The Antennapedia homeodomain-derived peptide penetratin was imported about three times as efficient as the basic fibroblast growth factor derived MTS peptide. Both peptides equilibrated between cytoplasm and nucleus. A relatively high mobility of these molecules inside the cells indicated that they may be rapidly degraded by cytosolic proteases. Based on these results, it will be possible to determine intracellular concentrations of inhibitors linked to import peptides directly by FCM at nanomolar concentrations and to optimise such constructs for proteolytic stability. PMID- 11934268 TI - Physico-chemical properties of the N-terminally truncated L68Q cystatin C found in amyloid deposits of brain haemorrhage patients. AB - Cystatin C, a major extracellular cysteine proteinase inhibitor, is deposited as amyloid in brain haemorrhage patients with hereditary cystatin C amyloid angiopathy (HCCAA). A disease-causing mutation on the genetic level results in the substitution Leu68-->Gln (L68Q) in cystatin C, which causes protein instability. Besides carrying the L68Q substitution, cystatin C in amyloid deposits isolated from patients is N-terminally truncated by 10 amino acids. To elucidate the role of the N-terminal truncation for protein stability and aggregation properties, (delta1-10,L68Q)-cystatin C was produced in an Escherichia coli expression system and characterised. Unlike wild-type cystatin C, this variant rapidly dimerised under physiological conditions. Two unfolding intermediates of (delta1-10,L68Q)-cystatin C were identified, under the same pH and ionic strength conditions as required to form intermediates of full-length L68Q cystatin C. No evidence was found that the N-terminal truncation per se alters protein stability and leads to higher forms of aggregation. Monomeric as well as dimeric L68Q cystatin C incubated with neutrophil elastase was truncated as in HCCAA patients' amyloid. A protein variant with a thrombin cleavage site placed in front of residue Gly11 in L68Q cystatin C was constructed and used to confirm that the N-terminal segment is similarly accessible to proteinases in the monomeric and dimeric states of L68Q cystatin C. Thus, the N-terminal segment of L68Q cystatin C is exposed to proteolytic attack and does not seem to be involved in intramolecular contacts leading to dimerisation or higher-order aggregation. We conclude that the N-terminal truncation likely is an event secondary to amyloid formation, and of no relevance for the development of HCCAA. PMID- 11934269 TI - Characterization of the sialate-7(9)-O-acetyltransferase from the microsomes of human colonic mucosa. AB - Sialic acids present on human colonic mucins are highly O-acetylated, however, little is known about the underlying enzymatic activity required for O acetylation in this tissue. Here we report on the substrate specificity, subcellular localization and characterization of the sialate-7(9)-O acetyltransferase in normal human colonic mucosa. Using CMP-Neu5Ac, the most efficient acceptor substrate of all those tested, the enzymatic activity was found to be optimal at 37 degrees C, with a pH optimum of 7.0. Activity was also found to be dependent on protein, CMP-Neu5Ac (Km: 59.2 microM) and AcCoA (Km: 6.1 microM) concentrations, as well as membrane integrity. The enzyme's activity could be inhibited by CoA with a Ki of 11.9 microM. In addition, enzymatic activity was found to be localized in the Golgi-enriched membrane fraction. The nature of the O-acetylated products formed were verified with the aid of chromatographic and enzymatic techniques. The main product was 9-O-acetylated Neu5Ac, with a significant amount of oligo-O-acetylated Neu5Ac also being detected. The utilization of CMP-Neu5Ac as the acceptor substrate was confirmed by the isolation and characterization of the putative product, CMP-Neu5,9Ac2, using ion-exchange chromatography. The ability of CMP-Neu5,9Ac2 to act as a sialic acid donor for sialyltransferases represents the conclusive demonstration for the formation of CMP-Neu5,9Ac2. PMID- 11934270 TI - Functional analysis of the Escherichia coli molybdopterin cofactor biosynthesis protein MoeA by site-directed mutagenesis. AB - Five moeA mutants were generated by replacing some conserved amino acids of MoeA by site-directed mutagenesis. The mutants were assayed for the ability to restore in vivo nitrate reductase activity of the moeA mutant Escherichia coli JRG97 and in vitro Neurospora crassa nit-1 nitrate reductase activity. The replacements Asp59AlaGly60Ala, Asp259Ala, Pro298AlaPro301Ala abolished the function of MoeA in Mo-molybdopterin formation and stabilization, reflected in the inability to restore nitrate reductase activity. The replacements Gly251AlaGly252Ala reduced, and that of Pro283Ala had no effect, on nitrate reductase activity. E. coli JRG97 cells transformed with mutants that failed to restore nitrate reductase activity showed by HPLC analysis a decreased level of molybdopterin-derived dephospho FormA as compared to bacteria transformed with wild-type moeA. The effects of the amino acid replacements on MoeA function may be explained in correlation with the MoeA crystal structure. PMID- 11934271 TI - (6R)-5,6,7,8-tetrahydro-L-monapterin from Escherichia coli, a novel natural unconjugated tetrahydropterin. AB - The structure of the major tetrahydropterin in Escherichia coli was determined as (6R)-5,6,7,8-tetrahydro-L-monapterin, i. e. (6R)-2-amino-5,6,7,8-tetrahydro-6 [(1S,2S)-1,2,3-trihydroxypropyl]pteridin-4(3H)-one. Although the stereochemical structure of the trihydroxypropyl side chain has been determined previously by fluorescence detected circular dichroism analysis on its aromatic derivative, the most important configuration at C(6) has not been clarified. The major difficulties for the determination of the chirality were instability toward air oxidation and very low concentration of the tetrahydropterin derivative. In the present study, the C(6)-configuration was determined as R by comparing its stable hexaacetyl derivative with authentic (6R)- and (6S)-hexaacetyl-5,6,7,8-tetrahydro L-monapterins by high performance liquid chromatography (HPLC) and HPLC-mass spectrometry (LC-MS). (6R)-5,6,7,8-Tetrahydro-L-monapterin is a new unconjugated tetrahydropterin from natural sources. PMID- 11934272 TI - The effect of spermine on plasmid condensation and dye release observed by fluorescence correlation spectroscopy. AB - We demonstrate that fluorescence correlation spectroscopy (FCS) can be employed to follow the conformational changes of DNA molecules induced by the addition of a cationic condensing compound (spermine). In our experiments the plasmid pHbetaAPr-1-neo (10 kbp; contour length 3.4 microm) was labeled with propidium iodide (PrIo) and then titrated with spermine to induce its condensation. When spermine was applied at concentrations above 5 microM (spermine/DNAphosphate=0.375), the diffusion time of the labeled plasmid dropped from 15 ms down to 3 ms (its diffusion coefficient, D, increased from 1.0x10(-12) m2/s to 6.0x10(-12) m2/s). The application of spermine was also accompanied by decreasing count rate and particle number, reflecting the dye's dissociation. The data presented show that FCS may become a valuable tool in studying supramolecular aggregate formation, especially when association is followed by a change in the hydrodynamic size of the resulting complex. PMID- 11934273 TI - Muscovite (mica) allows the characterisation of supported bilayers by ellipsometry and confocal fluorescence correlation spectroscopy. AB - We demonstrate for the first time that ellipsometry and confocal fluorescence correlation spectroscopy (FCS) are complementary methods for the characterisation of supported planar phospholipid bilayers (SPBs) formed on mica, a mineral used in atomic force microscopy investigations of SPBs. Addition of small unilamellar vesicles containing 20% dioleoyl-phosphatidylserine (DOPS) and 80% dioleoyl phosphatidylcholine (DOPC) to an oxidised borosilicate surface, on the other hand, results in a planar lipid system characterised by lateral diffusion coefficients which are three time smaller than those obtained for SPBs. Moreover, seven labelled phospholipids were tested for their suitability in the FCS characterisation of vesicles as well as of SPBs. PMID- 11934274 TI - Involvement of carboxyl and phenoxyl groups in Cajanus cajan lectin-sugar interaction: interpretation of titration curves. AB - Hydrogen ion titration of an affinity-purified mannose/glucose-specific lectin from Cajanus cajan seeds at 30 degrees C and ionic strength of 0.15 was reported earlier by our lab (Salahuddin and Khan, 1998). Further work has been carried out on the systematic study of H+ titration of Cajanus cajan lectin at three different temperatures, i.e., 10 degrees, 20 degrees, and 30 degrees C at ionic strength of 0.15. In all, 88 protons were dissociated reversibly in pH range 2 12. Different ionizable groups present in lectin were also determined with respect to their heat of ionization. Heat of ionization, deltaH, for 43 carboxyl groups was 1.92 Kcal/mol, and for 7 phenoxyl groups, 6.84 Kcal/mol. Spectrophotometric titration was performed in the presence and absence of 0.1 M methyl alpha-D-mannopyranoside to characterize the role of phenoxyl groups present at the active site. One phenoxyl group was found to be involved in carbohydrate binding. The presence of two carboxyl groups per subunit at the active site was confirmed by H+ titration in the presence and absence of 0.1 M methyl alpha-D-mannopyranoside. Hemagglutinating and IgM precipitating activity was checked for the chemically modified carboxyl and phenoxyl group preparation. Loss of activity further confirmed the participation of carboxyl as well as phenoxyl groups at the active site. It is interesting to note the near-UV CD changes in the aromatic environment/conformation of the lectin in the presence of specific sugar, further indicating the possible participation of phenoxyl groups in the carbohydrate-binding pocket. PMID- 11934275 TI - Purification and characterization of porcine testis 90-kDa heat shock protein (HSP90) as a substrate for various protein kinases. AB - We purified a large quantity of HSP90 from porcine testis by hydroxylapatite (HA HSP90) and SDS-PAGE/electroelution (eluted-HSP90) to explore the molecular mechanism of HSP90 phosphorylation affecting its metabolism. The purified HSP90 was used as an antigen to raise polyclonal antibodies in rabbits. Immunoblot analysis revealed that most purified HSP90 was HSP90alpha. Compared with the commercial anti-HSP90 antibody, the polyclonal antibody raised in this study could specifically detect the testis HSP90 and immunoprecipitate HSP90 from tissue homogenates or cell extracts. Incubation of the purified HSP90 or HSP90 immunoprecipitated from extracts of human A431 cells, Balb/c 3T3 fibroblasts, and porcine testis with [gamma-32P]ATP/Mg2+ resulted in phosphorylation of HSP90. However, the eluted-HSP90 lost its phosphorylation ability when incubated with [gamma-32P]ATP x Mg2+ alone but could be phosphorylated by various protein kinases, including PKA, CKII, kinase FA/GSK-3 alpha, and AK. The order of phosphorylation of HSP90 by these kinases is PKA = CKII > AK >> kinase FA/GSK-3 alpha. PMID- 11934276 TI - Terbium(III) fluorescence probe studies on metal ion-binding sites in anticoagulation factor I from Agkistrodon acutus venom. AB - Anticoagulation factor I (ACF I) isolated from the venom of Agkistrodon acutus is an activated coagulation factor X-binding protein with marked anticoagulant activity. Present studies show that holo-ACF I assumes a compactly folded structure in the range of pH 5-6, in which the most interior Trp residues and quenchers are adjacent. Tb3+ ions can completely replace both Ca2+ ions in holo ACF I, as determined by equilibrium dialysis. Although the two Tb3+ ions in Tb3+ ACF I have slightly different luminescence efficiencies, both have similar quenching effects on the intrinsic fluorescence, suggesting that probably there are same numbers of Trp residues close to both Tb3+-binding sites. Two Tb3+ binding sites with similar apparent Tb3+ association constant values, (1.69 +/- 0.02) x 10(7) M(-1) and (1.42 +/- 0.01) x 10(7) M(-1), respectively, were further identified through Tb3+ fluorescence titration. In addition, it has been confirmed from the titration of holo-ACF I and Tb3+-ACF I with NBS that only interior Trp residues are involved in the energy transfer to Tb3+ ions and that all accessible Trp residues located in the surface of holo-ACF I have similar affinity to NBS, while those located in the surface of Tb3+-ACF I have two different kinds of affinity to NBS, which strongly suggests a conformational change of holo-ACF I upon substitution of Tb3+ for Ca2+. The results show that although the Tb3+-altered conformation of ACF I cannot support the binding of Tb3+-ACF I with FXa, determined by nondenaturing PAGE, Tb3+ ions are effective and useful fluorescence probes to analyze the structures and properties of Ca2+ binding sites in ACF I. PMID- 11934277 TI - Urea-induced denaturation of human serum albumin labeled with acrylodan. AB - We induced the denaturation of unlabeled human serum albumin (HSA) and of similar albumin labeled with acrylodan (6-acryloyl-2-dimethylamino naphthalene) with urea and studied the transition profiles using circular dichroism and fluorescence spectroscopy. The circular dichroism spectra for both albumin preparations resulted in the same curves, thus indicating that labeling with acrylodan does not perturb the conformation of HSA. Our results indicate that the denaturation of both albumin preparations takes place at a single, two-state transition with midpoint at about 6 M urea, due to the unfolding of its domain II. It is important to point out that even at 8 M urea, some residual structure remains in the HSA. Great changes in the fluorescence of the dye bound to the protein were observed by addition of solid guanidine hydrochloride to the protein labeled with acrylodan dissolved in 8 M urea, indicating that domain I of this protein was not denatured by urea. PMID- 11934278 TI - Separation and structure-function studies of Taiwan cobra cardiotoxins. AB - Six cardiotoxins (CTXs) and one cardiotoxin-like basic protein (CLBP) from Naja naja atra (Taiwan cobra) venom were separated by a SP-Sephadex C-25 column. CTXn and CTXI were well separated by eluting with ammonium acetate buffer, and the separation of CLBP from CTXIV and CTXV mixtures was achieved using sodium phosphate buffer. These findings suggest a differential interaction of CTXs with the chromatographic matrix using different buffer systems. Chemical modification studies on cationic residues of CTXI suggested that there was no single lysine or arginine residue exclusively responsible for its biological activity. Moreover, it was found that the cytotoxicity and hemolytic sites of CTXI could be dissociated by chemical modifications. It suggests the potentiality for preparing toxin derivatives in which a specific activity is retained. PMID- 11934279 TI - Effect of trifluoroethanol on the structural and functional properties of alpha crystallin. AB - Alpha crystallin is an eye lens protein with a molecular weight of approximately 800 kDa. It belongs to the class of small heat shock proteins. Besides its structural role, it is known to prevent the aggregation of beta- and gamma crystallins and several other proteins under denaturing conditions and is thus believed to play an important role in maintaining lens transparency. In this communication, we have investigated the effect of 2,2,2-trifluoroethanol (TFE) on the structural and functional features of the native alpha-crystallin and its two constituent subunits. A conformational change occurs from the characteristic beta sheet to the alpha-helix structure in both native alpha-crystallin and its subunits with the increase in TFE levels. Among the two subunits, alphaA crystallin is relatively stable and upon preincubation prevents the characteristic aggregation of alphaB-crystallin at 20% and 30% (v/v) TFE. The hydrophobicity and chaperone-like activity of the crystallin subunits decrease on TFE treatment. The ability of alphaA-crystallin to bind and prevent the aggregation of alphaB-crystallin, despite a conformational change, could be important in protecting the lens from external stress. The loss in chaperone activity of alphaA-crystallin exposed to TFE and the inability of peptide chaperone--the functional site of alphaA-crystallin--to stabilize alphaB crystallin at 20-30% TFE suggest that the site(s) involved in subunit interaction and chaperone-like function are quite distinct. PMID- 11934280 TI - Aminoglycosides as substrates and inhibitors of peroxidases: a possible role of these antibiotics against myeloperoxidase-dependent cytotoxicity. AB - The kinetics of the catalytic cycle of myeloperoxidase and of horseradish peroxidase reacting with aminoglycosides have been studied by conventional and stopped-flow spectrophotometry. Aminoglycosides acted as one-electron reducing substrates converting compound I, formed when stoichiometric amounts of hydrogen peroxide were added to the enzyme, to compound II, and compound II to the resting, ferric enzyme. The latter gradually decayed into a further spectroscopic derivative (lambda(max) = 540 and 403 nm) tentatively identified as a complex of ferric heme with the antibiotic oxidation product(s), and the resulting enzyme was fully inactivated. Since myeloperoxidase is the only human enzyme known to convert chloride ions into the cytotoxic hypochlorous acid, the data presented in this paper bear relevance to the pharmacological effects of aminoglycoside antibiotics, which, while inhibiting bacterial growth, also prevent oxidative cellular damage caused by hypochlorous acid aging as substrates and inhibitors of myeloperoxidase. PMID- 11934281 TI - Reciprocal 13C-labeling: a method for investigating the catabolism of cosubstrates. AB - The principle of reciprocal labeling is to use a uniformly 13C-labeled substrate as the primary carbon source and a naturally labeled cosubstrate. Metabolites derived from a naturally labeled cosubstrate, in this case amino acids, can then be identified by their relatively lower content of 13C, and information on the degradation pathway can be deduced. The technique is based on GC-MS measurements of amino acid labeling patterns, making the technique well suited for investigating the relative importance of amino acid biosynthesis and amino acid uptake from the medium, as the 13C content of the amino acids incorporated into biomass is a direct measure of the amino acid biosyntheses. The technique is illustrated by the investigation of the degradation of phenoxyacetic acid, a medium component that is essential for production of penicillin V by Penicillium chrysogenum. Glucose was used as the uniformly labeled primary carbon source. PMID- 11934282 TI - Combined effects of temperature and medium composition on exopolysaccharide production by Lactobacillus rhamnosus RW-9595M in a whey permeate based medium. AB - The effects of temperature (22-42 degrees C), whey permeate concentration (WP, 1.6-8.4%), and supplementation level with yeast nitrogen base (YNB, 0-2.0%) on exopolysaccharide (EPS) production was studied during 20 pH-controlled (pH = 6.0) batch cultures with Lactobacillus rhamnosus RW-9595M, using a central composite design (CCD). The EPS production was measured using both the conventional method based on ethanol precipitation of EPS and a new ultrafiltration (UF) method. EPS production was not growth-associated for high temperatures (32-42 degrees C) and WP concentrations (7.0-8.4%). In contrast, at suboptimal temperature (22-26 degrees C), EPS production was growth-associated. Maximal EPS production measured with the UF method was approximately 2-fold higher than those measured with the conventional method and varied from 125 to 477 mg/L. This parameter was significantly influenced by WP and YNBWP interaction, whereas ANOVA for maximal EPS production measured by the conventional method did not show significant factor effects. EPS volumetric productivities varied from 3.0 to 16.4 mg EPS/L small middle doth. YNB supplementation did not promote cell growth but did increase EPS production at high WP concentrations. Our data indicate the potential of L. rhamnosus RW-9595M for producing EPS in a supplemented WP medium and suggest that this production could be further increased by the addition of a growth-limiting nutrient in the medium. PMID- 11934283 TI - Prediction of naphthalene bioaccumulation using an adipocyte cell line model. AB - A long-term goal of this research is to develop an in vitro model to study the metabolism, distribution, and fate of chemicals or pharmaceuticals in animals and humans. An important component of such a system is an in vitro model to study bioaccumulation of specific chemicals in adipose tissue. Due to the difficulties in maintaining primary adipocytes in culture and conducting reproducible experiments, transformed adipocyte cell lines have been used as an alternative. In this paper, several rodent preadipocyte cell lines (3T3-L1, 3T3-F442A, and TA1 cells) that differentiate into adipocytes when exposed to the appropriate stimuli are tested as an investigative tool to study naphthalene accumulation. The in vitro model is tested by comparison of its performance to that of primary adipocytes. All the experimental evidence supports the hypothesis that naphthalene accumulation is primarily dependent on the level of intracellular lipid. Furthermore, the level of naphthalene bioaccumulation is linearly correlated with the amount of triglyceride content with the slope of 37.7 +/- 0.5 microg of naphthalene/(mg of triglyceride). Indomethacin/dexamethasone/insulin are shown to be more effective in promoting preadipocyte differentiation than methylisobutylxanthine/dexamethasone/insulin. Additionally, external factors, such as the presence of albumin and serum in the medium, affect the cellular naphthalene uptake by decreasing the amount of naphthalene transported into fat cells. Among the three cell lines tested, 3T3-L1 adipocytes accumulated the highest intracellular lipid and, hence, yielded the highest level of naphthalene accumulation. Its ability to accumulate naphthalene is comparable to that of primary adipocytes. The 3T3-L1 adipocyte model is appropriate for studying the bioaccumulation of xenobiotics that are aromatic hydrocarbons. PMID- 11934284 TI - Development of mammalian serum albumin affinity purification media by peptide phage display. AB - Several phage isolates that bind specifically to human serum albumin (HSA) were isolated from disulfide-constrained cyclic peptide phage-display libraries. The majority of corresponding synthetic peptides bind with micromolar affinity to HSA in low salt at pH 6.2, as determined by fluorescence anisotropy. One of the highest affinity peptides, DX-236, also bound well to several mammalian serum albumins (SA). Immobilized DX-236 quantitatively captures HSA from human serum; mild conditions (100 mM Tris, pH 9.1) allow release of HSA. The DX-236 affinity column bound HSA from human serum with a greater specificity than does Cibacron Blue agarose beads. In addition to its likely utility in HSA and other mammalian SA purifications, this peptide media may be useful in the proteomics and medical research markets for selective removal of mammalian albumin from serum prior to mass spectrometric and other analyses. PMID- 11934285 TI - Modeling of growth, lactate consumption, and volatile fatty acid production by Megasphaera elsdenii cultivated in minimal and complex media. AB - Use of the Pirt and Luedeking-Piret equations permits the determination of the effect of medium composition on the metabolic patterns of Megasphaera elsdenii grown in minimal and complex media with lactate as the major carbon source. To establish the significance of the parameters involved in the Pirt and Luedeking Piret equations, a quantitative statistical criterion was proposed. In the complex medium, lactate was completely used for growth and product formation, whereas in the minimal medium a fraction of the energy obtained from lactate was used for maintenance purposes. Modeling of VFA production by the Luedeking-Piret equation showed that, independent of the type of medium, acetate and propionate are growth-associated products, while butyrate and valerate are only partially growth-associated. The growth-associated products are related to energy-yielding metabolism and the non-growth-associated products are related to the consumption of reducing equivalents. PMID- 11934286 TI - Benzene-free synthesis of adipic acid. AB - Strains of Escherichia coli were constructed and evaluated that synthesized cis,cis-muconic acid from D-glucose under fed-batch fermentor conditions. Chemical hydrogenation of the cis,cis-muconic acid in the resulting fermentation broth has also been examined. Biocatalytic synthesis of adipic acid from glucose eliminates two environmental concerns characteristic of industrial adipic acid manufacture: use of carcinogenic benzene and benzene-derived chemicals as feedstocks and generation of nitrous oxide as a byproduct of a nitric acid catalyzed oxidation. While alternative catalytic syntheses that eliminate the use of nitric acid have been developed, most continue to rely on petroleum-derived benzene as the ultimate feedstock. In this study, E. coli WN1/pWN2.248 was developed that synthesized 36.8 g/L of cis,cis-muconic acid in 22% (mol/mol) yield from glucose after 48 h of culturing under fed-batch fermentor conditions. Optimization of microbial cis,cis-muconic acid synthesis required expression of three enzymes not typically found in E. coli. Two copies of the Klebsiella pneumoniae aroZ gene encoding DHS dehydratase were inserted into the E. coli chromosome, while the K. pneumoniae aroY gene encoding PCA decarboxylase and the Acinetobacter calcoaceticus catA gene encoding catechol 1,2-dioxygenase were expressed from an extrachromosomal plasmid. After fed-batch culturing of WN1/pWN2.248 was complete, the cells were removed from the broth, which was treated with activated charcoal and subsequently filtered to remove soluble protein. Hydrogenation of the resulting solution with 10% Pt on carbon (5% mol/mol) at 3400 kPa of H2 pressure for 2.5 h at ambient temperature afforded a 97% (mol/mol) conversion of cis,cis-muconic acid into adipic acid. PMID- 11934287 TI - Quantitative screening of yeast surface-displayed polypeptide libraries by magnetic bead capture. AB - Magnetic bead capture is demonstrated here to be a feasible alternative for quantitative screening of favorable mutants from a cell-displayed polypeptide library. Flow cytometric sorting with fluorescent probes has been employed previously for high throughput screening for either novel binders or improved mutants. However, many laboratories do not have ready access to this technology as a result of the limited availability and high cost of cytometers, restricting the use of cell-displayed libraries. Using streptavidin-coated magnetic beads and biotinylated ligands, an alternative approach to cell-based library screening for improved mutants was developed. Magnetic bead capture probability of labeled cells is shown to be closely correlated with the surface ligand density. A single pass enrichment ratio of 9400 +/- 1800-fold, at the expense of 85 +/- 6% binder losses, is achieved from screening a library that contains one antibody displaying cell (binder) in 1.1 x 10(5) nondisplaying cells. Additionally, kinetic screening for an initial high affinity to low affinity (7.7-fold lower) mutant ratio of 1:95,000, the magnetic bead capture method attains a single-pass enrichment ratio of 600 +/- 200-fold with a 75 +/- 24% probability of loss for the higher affinity mutant. The observed high loss probabilities can be straightforwardly compensated for by library oversampling, given the inherently parallel nature of the screen. Overall, these results demonstrate that magnetic beads are capable of quantitatively screening for novel binders and improved mutants. The described methods are directly analogous to procedures in common use for phage display and should lower the barriers to entry for use of cell surface display libraries. PMID- 11934288 TI - Kinetics of the pectin methylesterase catalyzed de-esterification of pectin in frozen food model systems. AB - The pectin methylesterase (PME) catalyzed de-esterification of pectin was studied in four frozen food model systems based on sucrose, fructose, maltodextrin, and carboxymethylcellulose (CMC) in a temperature range from -24 to 20 degrees C, with the aim of elucidating the applicability of the theory of "food polymer science" on the kinetics. The rate substantially decreased around the glass transition temperature in the case of CMC, while very low rates were observed far above the glass transition temperature in the case of maltodextrin, fructose, and sucrose model systems. In general, the kinetics of this reaction was found to be influenced more by factors such as the characteristics of the component solutes, freeze concentration, the possible viscosity enhancement due to a particular combination of solutes, and the molecular size of the substrate molecule rather than the glass transition process. The Arrhenius equation described the temperature dependence of kinetics both in the liquid state of all the systems studied (r(2) > or = 0.97) and the glassy state of CMC (r(2) = 0.95). A clear break in the Arrhenius plot was observed as the temperature decreased to subfreezing temperatures. The Arrhenius equation could describe the kinetics reasonably well in the rubbery state for fructose and sucrose model systems (r(2) > 0.992). In the case of maltodextrin and CMC, the Arrhenius plots showed a slight curvature followed by a break at the glass transition temperature for CMC. The WLF equation with system-dependent coefficients better described the kinetics in the rubbery state of the CMC and part of the maltodextrin system. A linear relationship between the logarithm of the rate and T - Tg' described the kinetics in the sucrose as well as fructose model systems (r(2) = 0.9928 and 0.993, respectively). PMID- 11934289 TI - Improved cell growth in tobacco suspension cultures expressing Vitreoscilla hemoglobin. AB - Expression of the gene encoding bacterial hemoglobin (VHb) from Vitreoscilla has been previously used to improve recombinant cell growth and enhance product formation under microaerobic conditions, a common phenomenon in large-scale cultivations of bacteria. This technology has now been applied to tobacco suspension cultures. Tobacco suspension cultures have been generated from VHb expressing tobacco plants. Cell cultures were capable of producing an active hemoglobin. When grown in shake flasks, the cells did not show any lag-phase and exhibited improved cell growth, compared to controls carrying the parental plasmid. PMID- 11934290 TI - Production of fructose and ethanol from sugar beet molasses using Saccharomyces cerevisiae ATCC 36858. AB - The production of enriched fructose syrups and ethanol from beet molasses using Saccharomyces cerevisiae ATCC 36858 was studied. In batch experiments with a total sugar concentration between 94.9 and 312.4 g/L, the fructose yield was above 93% of the theoretical value. The ethanol yield and volumetric productivity in the beet molasses media with sugar concentration below 276.2 g/L were in the range of 59-76% of theoretical value and between 0.48 and 2.97 g of ethanol/(L x h), respectively. The fructose fraction in the carbohydrates content of the produced syrups was more than 95% when the total initial sugar concentration in the medium was below 242.0 g/L. Some oligosaccharides and glycerol were also produced in all tested media. Raffinose and the produced oligosaccharides were completely consumed by the end of the fermentation process when the total initial sugar concentration was below 190.1 g/L. The glycerol concentration was below 16.1 g/L. The results could be useful for a potential industrial production of ethanol and high-fructose syrup from sugar beet molasses. PMID- 11934291 TI - Immobilization of beta-galactosidase on fibrous matrix by polyethyleneimine for production of galacto-oligosaccharides from lactose. AB - The production of galacto-oligosaccharides (GOS) from lactose by Aspergillus oryzae beta-galactosidase immobilized on cotton cloth was studied. A novel method of enzyme immobilization involving PEI-enzyme aggregate formation and growth of aggregates on individual fibrils of cotton cloth leading to multilayer immobilization of the enzyme was developed. A large amount of enzyme was immobilized (250 mg/g support) with about 90-95% efficiency. A maximum GOS production of 25-26% (w/w) was achieved at near 50% lactose conversion from 400 g/L of lactose at pH 4.5 and 40 degrees C. Tri- and tetrasaccharides were the major types of GOS formed, accounting for about 70% and 25% of the total GOS produced in the reactions, respectively. Temperature and pH affected not only the reaction rate but also GOS yield to some extend. A reaction pH of 6.0 increased GOS yield by as much as 10% compared with that of pH 4.5 while decreased the reaction rate of immobilized enzyme. The cotton cloth as the support matrix for enzyme immobilization did not affect the GOS formation characteristics of the enzyme under the same reaction conditions, suggesting diffusion limitation was negligible in the packed bed reactor and the enzyme carrier. Increase in the thermal stability of PEI-immobilized enzyme was also observed. The half-life for the immobilized enzyme on cotton cloth was close to 1 year at 40 degrees C and 21 days at 50 degrees C. Stable, continuous operation in a plug-flow reactor was demonstrated for about 3 days without any apparent problem. A maximum GOS production of 26% (w/w) of total sugars was attained at 50% lactose conversion with a feed containing 400 g/L of lactose at pH 4.5 and 40 degrees C. The corresponding reactor productivity was 6 kg/L/h, which is several-hundred-fold higher than those previously reported. PMID- 11934292 TI - Transformation of thiodiglycol by resting cells of Alcaligenes xylosoxydans PGH10. AB - A new strain of Alcaligenes xylosoxydans able to aerobically cometabolize thiodiglycol, the primary hydrolysis product of sulfur mustard, was isolated and tested in a laboratory scale stirred tank reactor. The strain, named PGH10, cannot use TDG as sole carbon and energy source for growth, but resting cells previously grown on either rich broth or defined mineral media efficiently metabolize this compound through [(2-hydroxyethyl)thio]acetic acid and thiodiacetic acid as intermediates. Degradation of TDG by PGH10 is shown to take place at late exponential and stationary phase but is not triggered by carbon exhaustion. Cultures pregrown to saturation for 48 h in the absence of TDG can be stored and used for degradation of TDG, reducing significantly the time required to achieve the reduction of the compound concentration to undetectable levels. Degradation can take place in buffered media with no carbon source added, although best results were obtained in mineral media supplemented with citrate or fructose. Oxidation to [(2-hydroxyethyl)thio]acetic acid and thiodiacetic acid was proposed to be catalyzed by a butanol-dehydrogenase activity. Inhibition of TDG transformation in the presence of several alcohols is also shown. PMID- 11934294 TI - An efficient enzymatic Baeyer-Villiger oxidation by engineered Escherichia coli cells under non-growing conditions. AB - Economical methods of supplying NADPH must be developed before biotransformations involving this cofactor can be considered for large-scale applications. We have studied the enzymatic Baeyer-Villiger oxidation of cyclohexanone as a model for this class of reactions and developed a simple approach that uses whole, non growing Escherichia coli cells to provide high productivity (0.79 g epsilon caprolactone/L/h = 18 micromol epsilon-caprolactone/min/g dcw) and an 88% yield. Glucose supplied the reducing equivalents for this process, and no exogenous cofactor was required. The volumetric productivity of non-growing cells was an order of magnitude greater than that achieved with growing cells of the same strain. Cells of an engineered E. coli strain that overexpresses Acinetobacter sp. cyclohexanone monooxygenase were grown under inducing conditions in rich medium until the entry to stationary phase; the subsequent cyclohexanone oxidation was carried out in minimal salts medium lacking a nitrogen source. After the biotransformation was complete, the lactone product was adsorbed to a solid support and recovered by washing with an organic solvent. PMID- 11934293 TI - Purification and identification of an Escherichia coli beta-keto ester reductase as 2,5-diketo-D-gluconate reductase YqhE. AB - An NADPH-dependent enzyme that reduces ethyl 2-methylacetoacetate stereoselectively to ethyl (2R)-methyl-(3S)-hydroxybutanoate was purified 730 fold from Escherichia coli. The N-terminal amino acid sequence data obtained from the purified reductase were used to search the E. coli genome, and a single match was found at the start of the yqhE open reading frame. The YqhE protein had been identified previously by Yum et al. as a 2,5-diketo-D-gluconate reductase on the basis of sequence similarity to other bacterial homologues [Yum, D.-Y.; Lee, B. Y.; Pan, J.-G. Appl.Environ. Microbiol. 1999, 65, 3341-3346]; however, it had not been examined for beta-keto ester reductions. Our results thus link a key enzyme in the microbial production of ascorbate with stereoselective beta-keto ester reductions, two important fields in biocatalysis. The purified YqhE reductase accepts ethyl acetoacetate and a variety of 2-substituted derivatives, and its sequence is similar to other aldose reductase superfamily members that also reduce alpha-substituted beta-keto esters to syn-(2R,3S) alcohols. PMID- 11934295 TI - Automated production support for the bioprocess industry. AB - This paper describes the application of Artificial Intelligence and Multivariate Statistical Techniques to two industrial fermentation systems. In the first example, an Expert System is shown to provide tighter control of an important process parameter. This is shown to lead to improved consistency of operation. In the second application, Principal Component Analysis is applied to a final stage fermentation production facility. The results presented indicate that the algorithm can provide concise indicators of process faults that can be presented to the operators to assist them in taking suitable corrective actions. PMID- 11934296 TI - Coupling of surface carboxyls of carboxymethylcellulase with aniline via chemical modification: extreme thermostabilization in aqueous and water-miscible organic mixtures. AB - We wish to report the attainment of the highest ever T(opt) by introducing approximately two aromatic rings through chemical modification of surface carboxyl groups in carboxymethylcellulase from Scopulariopsis sp. with concomitant decrease in V(max), K(m), and optimum pH! This extraordinary enhancement in thermophilicity of aniline-coupled CMCase (T(opt) = 122 degrees C) by a margin of 73 degrees C as compared with the native enzyme (T(opt) = 49 degrees C) is the highest reported for any mesophilic enzyme that has been modified either through chemical modification or site-directed mutagenesis. It is also reported for the first time that aniline coupled CMCase (ACC) is simultaneously thermostable in aqueous as well as water-miscible organic solvents. The T(opt) of native CMCase and ACC were 25 and 90 degrees C, respectively, in 40% (v/v) aqueous dioxan. The modified enzyme was also stabilized against irreversible thermal denaturation. Therefore, at 55 degrees C, ACC had a half-life of 136 min as compared with native CMCase whose half-life was only 5 min. We believe that the reasons for this elevated thermostability and thermophilicity are surface aromatic-aromatic interactions and aromatic interactions with the sugar backbone of the substrate, respectively. PMID- 11934297 TI - Effect of elicitation on growth, respiration, and nutrient uptake of root and cell suspension cultures of Hyoscyamus muticus. AB - The elicitation of Hyoscyamus muticus root and cell suspension cultures by fungal elicitor from Rhizoctonia solani causes dramatic changes in respiration, nutrient yields, and growth. Cells and mature root tissues have similar specific oxygen uptake rates (SOUR) before and after the onset of the elicitation process. Cell suspension SOUR were 11 and 18 micromol O2/g FW x h for non-elicited control and elicited cultures, respectively. Mature root SOUR were 11 and 24 micromol O2/g FW x h for control and elicited tissue, respectively. Tissue growth is significantly reduced upon the addition of elicitor to these cultures. Inorganic yield remains fairly constant, whereas yield on sugar is reduced from 0.532 to 0.352 g dry biomass per g sugar for roots and 0.614 to 0.440 g dry biomass per g sugar for cells. This reduction in yield results from increased energy requirements for the defense response. Growth reduction is reflected in a reduction in root meristem (tip) SOUR, which decreased from 189 to 70 micromol O2/g FW x h upon elicitation. Therefore, despite the increase in total respiration, the maximum local oxygen fluxes are reduced as a result of the reduction in metabolic activity at the meristem. This distribution of oxygen uptake throughout the mature tissue could reduce mass transfer requirements during elicited production. However, this was not found to be the case for sesquiterpene elicitation, where production of lubimin and solavetivone were found to increase linearly up to oxygen partial pressures of 40% O2 in air. SOUR is shown to similarly increase in both bubble column and tubular reactors despite severe mass transfer limitations, suggesting the possibility of metabolically induced increases in tissue convective transport during elicitation. PMID- 11934299 TI - Ion exchange membrane bioreactor for selective removal of nitrate from drinking water: control of ion fluxes and process performance. AB - An ion exchange membrane bioreactor (IEMB), consisting of a monoanion permselective membrane dialyzer coupled to a stirred anoxic vessel with an enriched mixed denitrifying culture, has been studied for nitrate removal from drinking water. The influence of nitrate and chloride concentrations on the selectivity of nitrate transport in the IEMB process was investigated. With appropriate dosing of chloride ions to the IEMB biocompartment, it was possible to regulate the net bicarbonate flux in the system, thus maintaining the bicarbonate concentration in the treated water at the desired level. The latter was not possible to achieve in Donnan dialysis, operated as a single process in which, besides the lower nitrate removal efficiency found, bicarbonate was co extracted together with nitrate from the polluted water stream. Residual carbon source (ethanol) and nitrite were not detected in the treated water produced in the IEMB system. With a concentration of nitrate in the polluted water three times higher than the maximum contaminant level of 50 mg L(-1) allowed, the IEMB process was successfully operated for a period of 1 month before exceeding this limit. PMID- 11934298 TI - Chromate reduction in Shewanella oneidensis MR-1 is an inducible process associated with anaerobic growth. AB - Cr(VI) reduction was observed during tests with Shewanella oneidensis MR-1 (previously named S. putrefaciens MR-1) while being grown with nitrate or fumarate as electron acceptor and lactate as electron donor. From the onset of anoxic growth on fumarate, we measured a gradual and progressive increase in the specific Cr(VI) reduction rate with incubation time until a maximum was reached at late exponential/early stationary phase. Under denitrifying conditions, the specific Cr(VI) reduction rate was inhibited by nitrite, which is produced during nitrate reduction. However, once nitrite was consumed, the specific reduction rate increased until a maximum was reached, again during the late exponential/early stationary phase. Thus, under both fumarate- and nitrate reducing conditions, an increase in the specific Cr(VI) reduction rate was observed as the microorganisms transition from oxic to anoxic growth conditions, presumably as a result of induction of enzyme systems capable of reducing Cr(VI). Although Cr(VI) reduction has been studied in MR-1 and in other facultative bacteria under both oxic and anoxic conditions, a transition in specific reduction rates based on physiological conditions during growth is a novel finding. Such physiological responses provide information required for optimizing the operation of in situ systems for remediating groundwater contaminated with heavy metals and radionuclides, especially those that are characterized by temporal variations in oxygen content. Moreover, such information may point the way to a better understanding of the cellular processes used by soil bacteria to accomplish Cr(VI) reduction. PMID- 11934300 TI - Characterization of polycationic amino acids fusion systems for ion-exchange purification of cyclodextrin glycosyltransferase from recombinant Escherichia coli. AB - Fusion proteins with charged polycationic amino acid tails were constructed for the purpose of simple ion-exchange purification with high purity. A number of positively charged lysine and arginine tails were fused to the C-terminus of cyclodextrin glycosyltransferase (CGTase) derived from Bacillus macerans and expressed in Escherichia coli. The ionic binding forces provided by the tails allowed the selective recovery of CGTase from recombinant E. coli cell extracts, while CGTase by itself could not bind to the cation exchanger at neutral pH. The type of amino acids used and the length of the tail directly affected the purification factors. Most intracellular proteins of E. coli adsorbed on the cation exchanger could be removed by washing with 400 mM NaCl solution at pH 7.4, suggesting that a fusion partner suitable for purification purpose should be provided with high binding strength and the maintenance of adsorption by washing with NaCl solution. Among the fusion CGTases constructed, the CGTK10ase containing 10 lysine residues provided sufficiently high binding strength to allow purification to its homogeneity through simple ion-exchange chromatography. PMID- 11934301 TI - Optimal packing characteristics of rolled, continuous stationary-phase columns. AB - Rolled, continuous stationary phases were constructed by tightly rolling and inserting a whole textile fabric into a chromatography column. This work reports the column performance, in terms of plate height, void fraction, and resolution, of 10 cellulose-based fabrics. The relation between fabric structural properties of yarn diameter, fabric count, fabric compressibility, and column performance are quantitated. General requirements, including reproducibility of packing, for choosing fabrics to make a good SEC column are identified. This research showed that the packed columns have an optimal mass of fabric that minimizes plate height and maximizes resolution, in a manner that is consistent with chromatography theory. Mass of material packed is then an important column parameter to consider when optimizing columns for the rapid desalting of proteins. Proteins were completely separated from salt and glucose in less than 8 min at a pressure drop less than 500 psi on the rolled, continuous stationary phase columns. These results, together with stability and reproducibility, suggest potential industrial applications for cellulose-based rolled, continuous stationary-phase columns where speed is a key parameter in the production process. PMID- 11934302 TI - Modeling pore size distribution in cellulose rolled stationary phases. AB - Rolled stationary phases are fabrics (i.e., nonparticulate phases) that rapidly separate proteins from salts on the basis of size exclusion. Pore size and pore size distributions in the stationary phase determine how different size molecules distribute between the stationary and mobile phases in liquid chromatography columns. The potential for size exclusion chromatography by fabrics is not initially obvious because their interlaced structures are atypical for size exclusion supports. A simple logistic model fits the pore size distribution of a rolled stationary phase when pore sizes were measured using PEG, Dextran, D2O, glucose, and NaCl probes. When the fabric is treated with cellulase enzymes, the water-accessible pores uniformly decrease and peak retention is lower. The logistic function model captures this result and enables comparison of pore size distribution curves between enzyme-treated and untreated fabrics in rolled stationary phase columns. PMID- 11934303 TI - Dean vortex membrane microfiltration and diafiltration of rBDNF E. coli inclusion bodies. AB - Cross-flow microfiltration (CMF) and diafiltration were used to concentrate and purify recombinant Brain-Derived Neutrophic Factor (rBDNF) inclusion bodies from an E. coli cell suspension and a homogenized E. coli cell suspension (homogenate/lysate). Although these processes have been tested industrially in pilot scale with conventional linear membrane microfiltration modules, their performances were severely limited due to membrane fouling. The purpose of this work was to determine whether Dean vortex microfiltration with controlled centrifugal instabilities (Dean vortices produced in helical flow) could be used to improve filtration performance over that observed with conventional linear cross-flow microfiltration (CMF). For the microfiltration experiments with the feeds containing cell and homogenate suspensions, improvements in flux of about 50 and 70%, respectively, were obtained with the helical module as compared with that obtained with the linear module. For diafiltration with the homogenate suspension as feed, solute transport (as measured by mass) was from 100 to 40% higher after 40 and 100 min, respectively, with the helical module as compared with that obtained with the linear module. In the presence of the neutral surfactant, Tween 20, solute transport for diafiltration was at least 25 times higher during the first 10 min of operation and 100% higher after 300 min with the helical module as compared with that obtained with the linear module. Clearly, improved filtration performance, a purer and more concentrated product, and substantial savings can be expected with the new Dean vortex filters. PMID- 11934304 TI - High-level secretory expression of penicillin amidase from Providencia rettgeri in Saccharomyces cerevisiae: purification and characterization. AB - Heterologous production of the heterodimeric penicillin G amidase (PAC) from Providencia rettgeri was optimized in Saccharomyces cerevisiae. Several factors, including the effect of different growth and induction conditions, were identified to be critical for the enzyme overproduction and secretion. The PAC yield was significantly increased by more than 500-fold compared to that obtained in the native bacterium, and the recombinant enzyme was almost entirely secreted. Electrophoretic characterization of the secreted rPAC(Pr), which was purified over 20-fold by a combination of hydrophobic interaction and ion-exchange chromatography, demonstrated a microheterogeneity of the recombinant enzyme. The recombinant PAC(Pr) was further characterized in terms of specific activity, pH, and temperature profiles and kinetic parameters. The data presented here suggest that by overexpressing rPAC(Pr) in S.cerevisiae and purifying secreted enzyme from culture medium one can readily obtain a large amount of an alternative source of penicillin amidase with properties comparable to that of todays main industrial source of enzyme. PMID- 11934305 TI - Passaging protocols for mammalian neural stem cells in suspension bioreactors. AB - Mammalian neural stem cells (NSC) offer great promise as therapeutic agents for the treatment of central nervous system disorders. As a consequence of the large numbers of cells that will be needed for drug testing and transplantation studies, it is necessary to develop protocols for the large-scale expansion of mammalian NSC. Neural stem cells and early progenitor cells can be expanded in vitro as aggregates in controlled bioreactors using carefully designed media. The first objective of this study was to determine if it is possible to maintain a population of murine neural stem and progenitor cells as aggregates in suspension culture bioreactors over extended periods of time. We discovered that serial passaging of a mixture of aggregates sizes resulted in high viabilities, high viable cell densities, and good control of aggregate diameter. When the NSC aggregates were serially subcultured three times without mechanical dissociation, a total multiplication ratio of 2.9 x 10(3) was achieved over a period of 12 days, whereas the aggregate size was controlled (mean diameter less than 150 microm) below levels at which necrosis would occur. Moreover, cell densities of 1.0 x 10(6) cells/mL were repeatedly achieved in batch culture with viabilities exceeding 80%. The second objective was to examine the proliferative potential of single cells shed from the surface of these aggregates. We found that the single cells, when subcultured, retained the capacity to generate new aggregates, gave rise to cultures with high viable cell densities and were able to differentiate into all of the primary cell phenotypes in the central nervous system. PMID- 11934306 TI - Hyperosmotic stress and elevated pCO2 alter monoclonal antibody charge distribution and monosaccharide content. AB - Medium osmolality increases with pCO2 at constant pH. Elevated pCO2 and osmolality inhibit hybridoma growth to similar extents in both serum-containing and serum-free media. The combination of osmolality and elevated pCO2 synergizes to negatively impact cell growth. IgG2a glycosylation by hybridoma cells was evaluated under elevated pCO2 (to 250 mmHg pCO2) and/or osmolality (to 476 mOsm/kg). IgG2a site occupancy did not change significantly under any of the conditions studied, which is consistent with the robust glycosylation of other antibodies produced under various environmental stresses. However, changes were observed in the IgG2a charge distribution. Changes in the isoelectric point (pI) were greater under hyperosmotic stress, increasing by 0.32 and 0.41 pH units at 435 mOsm/kg in serum-containing and serum-free medium, respectively. Hyperosmotic stress also resulted in a concomitant increase in the heterogeneity of the charge distribution. The mean pI in serum-containing medium decreased by 0.16 pH units at 250 mmHg pCO2 when osmolality was controlled at 320 mOsm/kg but increased by 0.20 pH units when the osmolality increased with pCO2 (195 mmHg pCO2-435 mOsm/kg). In serum-free medium, elevated pCO2 did not alter pI, regardless of medium osmolality. In contrast to elevated osmolality at control pCO2, elevated pCO2 did not significantly alter the IgG2a charge heterogeneity under any of the conditions studied. The IgG2a was not sialylated, so sialylation changes were not responsible for changes in the charge distribution. IgG2a galactose content decreased with elevated osmolality, as a result of either elevated NaHCO3 or NaCl. However, when osmolality was controlled at elevated pCO2, the galactose content tended to increase. The mannose content decreased with increasing stress, while the fucose content remained relatively unchanged. It is likely that the observed increases in the pI of murine IgG2a were due to increased organellar pH, which is reflected by increased specific beta-galactosidase activity in the supernatant. PMID- 11934307 TI - Modeling of cryopreservation of engineered tissues with one-dimensional geometry. AB - Long-term storage of engineered bio-artificial tissues is required to ensure the off-the-shelf availability to clinicians due to their long production cycle. Cryopreservation is likely the choice for long-term preservation. Although the cryopreservation of cells is well established for many cell types, cryopreservation of tissues is far more complicated. Cells at different locations in the tissue could experience very different local environmental changes, i.e., the change of concentration of cryoprotecting chemicals (CPA) and temperature, during the addition/removal of CPA and cooling/warming, which leads to nonuniformity in cell survival in the tissue. This is due to the limitation of mass and heat transfer within the tissue. A specific aim of cryopreservation of tissue is to ensure a maximum recovery of cells and their functionality throughout a tissue. Cells at all locations should be protected adequately by the CPA and frozen at rates conducive to survival. It is hence highly desirable to know the cell transient and final states during cryopreservation within the whole tissue, which can be best studied by mathematical modeling. In this work, a model framework for cryopreservation of one-dimensional artificial tissues is developed on the basis of solving the coupled equations to describe the mass and heat transfer within the tissue and osmotic transport through the cell membrane. Using an artificial pancreas as an example, we carried out a simulation to examine the temperature history, cell volume, solute redistribution, and other state parameters during the freezing of the spherical heterogeneous construct (a single bead). It is found that the parameters affecting the mass transfer of CPA in tissue and through the cell membrane and the freezing rate play dominant roles in affecting the cell volume transient and extracellular ice formation. Thermal conductivity and extracellular ice formation kinetics, on the other hand, have little effect on cell transient and final states, as the heat transfer rate is much faster than mass diffusion. The outcome of such a model study can be used to evaluate the construct design on its survivability during cryopreservation and to select a cryopreservation protocol to achieve maximum cell survival. PMID- 11934308 TI - Development of a bioartificial nerve graft. I. Design based on a reaction diffusion model. AB - A simple reaction-diffusion model has been developed to describe the mass transport of nutrients and nerve growth factor within a bioartificial nerve graft (BNG). The BNG consists of a porous polymer conduit that is preseeded with Schwann cells in its lumen. The Schwann cells produce growth factors to stimulate nerve regeneration within the lumen of the conduit. The model can predict the wall thickness, porosity, and Schwann cell seeding density needed to maximize the axon extension rate while ensuring that sufficient nutrients, especially oxygen, are made available to the neurons until the formation of the neovasculature. The model predicts a sixteen-fold increase in the levels of nerve growth factor by dropping the porosity from 95 to 55% but only at the expense of reducing the oxygen concentration. At higher porosities, increasing the wall thickness and increasing the Schwann cell seeding density both have the same effect of increasing the concentration of nerve growth factor within the lumen of the conduit. This model provides a simple tool for evaluating various conduit designs before continuing with experimental studies in vivo. PMID- 11934309 TI - Development of a bioartificial nerve graft. II. Nerve regeneration in vitro. AB - A promising alternative for the repair of peripheral nerve injuries is the bioartificial nerve graft, or BNG, comprised of a tubular conduit preseeded with Schwann cells, which are an effective substrate for enhancing nerve regeneration. The physical properties of the conduit, porosity and wall thickness, as well as the Schwann cell seeding density, were tested for their effect on axon growth using rat dorsal root ganglia. These parameters can influence the amount of nutrients and growth factors made available to the neural tissue. Results show that a greater wall thickness and lower porosities have a detrimental effect on the growth of the axons. Over a four week period, axons extended 3.2 mm for the optimum case (DeltaR = 0.82 mm, epsilon = 0.75) compared to 1.8 and 1.6 mm for a lower porosity (0.55) and a greater wall thickness (1.4 mm), respectively. A maximum in the growth rate occurs at a porosity of 75% for Schwann cell seeded conduits but not for unseeded ones. When compared to mass transfer predictions, the results suggest that, at higher porosities, more growth factors diffuse out of the conduit, while at low porosities there is competition for nutrients. Increasing the Schwann cell seeding density enhances growth but also leads to an increase in the number of axons along the length of the conduit. This is indicative of branching of the axons, which requires additional resources to maintain and can lead to painful neuroma formation. Wall thickness and porosity were found not to have any significant effect on the axon number sprouting from the dorsal root ganglia and the mean diameter (p > 0.05). Considerations need to be made, not just on the polymer used, but also on its porosity, wall thickness, and Schwann cell seeding density. These parameters can be adjusted to create a bioartificial nerve graft that provides the optimal environment for nerve growth. PMID- 11934310 TI - Assessment of the performance of a fed-batch cultivation from the preculture quality using an electronic nose. AB - An electronic nose, a gas-phase multisensor system, was used to monitor precultivations of a recombinant tryptophan-producing Escherichia coli strain. The electronic nose signals showed a high correlation toward the main stages of the precultivations, namely, exponential growth, oxygen-limited growth, and glucose depletion. Principal component analysis (PCA) of the electronic nose signals was performed and shown to be useful for monitoring preculture progression. More importantly, PCA also allowed a qualitative assessment of the preculture performance during subsequent fed-batch cultivations. The electronic nose signals from the precultures showed, furthermore, a high correlation to the time of phosphate limitation and the tryptophan yield coefficient of the subsequent fed-batch cultivations, which allowed an accurate prediction of these process variables using partial least squares (PLS). The results demonstrate on data from 12 cultivations how the electronic nose can be a useful tool for the assessment of inoculum quality, thereby providing means of reducing batch-to batch variation and increasing the productivity of bioprocesses. PMID- 11934311 TI - Use of at-line spectrophotometry for the rapid definition of pilot-scale flocculation processes. AB - Traditionally most downstream bioprocesses have been operated without real-time knowledge of product and key contaminants, yielding little confidence in their operation and the impact on subsequent operations. A rapid UV-vis spectral prediction technique has been successfully demonstrated for the at-line characterization of a large scale continuous flocculation process in terms of RNA, key protein contaminants, and cell debris. A comparison was made between the spectral predictions and retrospective wet chemical assays, and a highly linear correlation was obtained. The spectral analysis technique allowed for real-time system information, which was applied to control the flocculation process to maintain satisfactory process performance, even when subjected to given possible process disturbances. PMID- 11934312 TI - Stringent regulation and high-level expression of heterologous genes in Escherichia coli using T7 system controllable by the araBAD promoter. AB - The recombinant Eschreichia coli strain BL21 (BAD) was constructed to carry a chromosomal copy of T7 gene 1 fused to the araBAD promoter. To further characterize this expression system, strain BL21 (BAD) was transformed with the plasmid containing the carbamoylase gene from Agrobacterium radiobacter driven by the T7 promoter. Upon induction with L-arabinose, recombinant cells produced 100 fold increase in carbamoylase activity in comparison with uninduced cells on M9 semidefined medium plus glycerol. This protein yield accounts for 30% of total cell protein content. In addition, it was found that after 100 generations the plasmid harboring the carbamoylase gene remained firmly stable in strain BL21 (BAD), but its stability dropped to only 20-30% in strain BL21 (DE3), a commercial strain bearing T7 gene 1 regulated by the lacUV5 promoter in its chromosome. In an attempt to enhance the total protein yield, fed-batch fermentation process was carried out using a two-stage feeding strategy to compartmentalize cell growth and protein synthesis. In the batch fermentation stage, the culture was grown on glucose to reach the stationary growth phase. Subsequently, glycerol was fed to the culture broth and L-arabinose was augmented to induce protein production when cells entered the late log growth phase. As a result, a carbamoylase yield corresponding to 5525 units was obtained, which amounts to a 337-fold increase over that achieved on a shake-flask scale. Taken together, these results illustrate the practical usefulness of T7 system under control of the araBAD promoter for heterologous protein production. PMID- 11934314 TI - Regulation (alteration) of activity and conformation of sucrase by coaggregation with cellobiase in culture medium of Termitomyces clypeatus. AB - Extracellular sucrase (S) of Termitomyces clypeatus was aggregated with cellobiase (C) in culture filtrate and coaggregates of sucrase to cellobiase with different activity ratios (S/C) were obtained during purification. Specific activity of the enzyme decreased significantly, after purification of sucrase free from cellobiase. Purified sucrase was characterized as a glycoprotein of molar mass around 55kDa as indicated by SDS-PAGE and HPGPLC. K(m) and V(max) of the purified enzyme were determined as 34.48 mM and 13.3 U/mg, respectively, at optimum temperature (45 degrees C) and pH (5.0). Substrate affinity and reaction velocity of the purified enzyme, free from cellobiase, was lowered by approximately 3.5 and 55 times, respectively, than that of the enzyme obtained from culture filtrate. The instant regain of sucrase activity up to the extent of 41% was obtained on in vitro addition of cellobiase (free from sucrase) to the enzyme in incubation mixture. Conformation of the enzyme free from cellobiase appeared to be significantly different from that of the coaggregate, as analyzed by circular dichroic and light scattering spectroscopy. It was concluded that activity and conformation of sucrase is regulated (altered) by heteroaggregation with cellobiase in the fungus. PMID- 11934313 TI - Alginate/aminopropyl-silicate/alginate membrane immunoisolatability and insulin secretion of encapsulated islets. AB - We utilized the sol-gel reaction to prepare an immunoisolatable membrane for a microcapsule-shaped bioartificial pancreas. The membrane, derived from two precursors, 3-(aminopropyl)trimethoxysilane (APTrMOS) and tetramethoxysilane (TMOS), was formed onto calcium-alginate gel beads via electrostatic interaction. The molecular weight cutoff point of less than 150 000 required for immunoisolation was achieved at molar ratios ([APTrMOS]/[TMOS]) ranging from 0.60 to 2.40 with the amount of APTrMOS fixed at 3.40 mmol/(10 mL of calcium alginate). When encapsulated in a membrane prepared at the molar ratio of 0.60, the islets contracted in volume and showed no response to stimulation by a high glucose concentration. However, islets in a membrane prepared at the molar ratio of 2.40 showed no contraction and responded to the glucose stimulation at almost the same level as free islets. These results demonstrated that the molar ratio of the precursors was a dominant factor affecting membrane permeability and the insulin secretion activity of the encapsulated islets. PMID- 11934315 TI - Purification of kappa-casien glycomacropeptide from sweet whey with undetectable level of phenylalanine. AB - Glycomacropeptide (GMP) found in sweet whey is a biologically active compound released from kappa-casein by the action of chymosin during cheese making. This study was undertaken to purify GMP from sweet whey as a research chemical on a laboratory scale. Glycomacropeptide was isolated from proteins and other non-GMP compounds by deproteinization with trichloroacetic acid and gel chromatography on Sephacryl S-200. The purified GMP accounted for 0.12% of dry sweet whey powder and contained 107.0, 50.9, 61.2 and 4.3 microg, respectively, of sialic acid, galactose, galactosamine and phosphorus per mg dry weight. The GMP was of high purity, with its amino acid composition showing undetectable levels of phenylalanine, tyrosine and arginine, the amino acids that do not occur in bovine GMP. On gel electrophoresis, the GMP showed a single broad band with an average mobility faster than that of carbonic anhydrase (molecular weight = 31 kDa). The purified GMP may be used as a standard glycopeptide in chromatography and electrophoresis and may also be used to test various known or unknown properties and biological activities of this compound. PMID- 11934317 TI - Inattention to nonsuperimposable midline symmetry causes wavefront analysis error. AB - BACKGROUND: The nonsuperimposable mirror-image symmetry of the body (enantiomorphism) is reflected in the wavefront error maps of eyes. Averaging the wavefront errors of right and left eyes has the potential to adversely affect correlations made between wavefront error and visual acuity or other factors. Not only are the results of past studies using Zernike terms suspected of being invalid, there is concern about possible errors in the algorithms used to create customized corneal ablations. OBJECTIVE: To compare the results of analysis with and without correction for enantiomorphism. METHODS: Fourteen TMS-1 corneal topographic maps from 7 patients having with-the-rule astigmatism in both corneas were selected for Zernike decomposition to 45 terms. The maps were distributed among 3 groups: 7 right eye maps, 7 left eye maps, and 7 left eye maps in which the topography was transposed about the vertical axial to correct for enantiomorphism (left eye-corrected). The wavefront error difference between the right and left eyes was compared with the difference between the right eyes and the left eyes in which enantiomorphism was corrected (right eye vs left eye corrected). The left eye wavefront error was then compared with the left eye wavefront error after correction (left eye vs left eye-corrected). RESULTS: Correcting for enantiomorphism produced a statisticially significant difference in the first 5 radial orders of Zernike terms (P=.02). Of the 45 Zernike terms analyzed, 7 terms were significantly different at the P<.05 level in the right eye vs left eye category, compared with 4 terms in the right eye vs left eye corrected category. Eleven terms were significantly different at the P<.05 level in the left eye vs left eye-corrected category. CONCLUSIONS: Correcting for enantiomorphism makes the Zernike terms in right and left eyes appear more similar. Failure to correct for enantiomorphism causes certain terms to cancel each other when averaged across right and left eyes. Wavefront error studies that do not consider enantiomorphism, including those used to adjust laser surgical nomograms, will introduce significant errors to certain Zernike terms. PMID- 11934316 TI - Clinical and theoretical results of intraocular lens power calculation for cataract surgery after photorefractive keratectomy for myopia. AB - OBJECTIVES: To describe the refractive results of cataract surgery after photorefractive keratectomy (PRK) for patients with myopia, and to find a more accurate method to predict intraocular lens (IOL) power in these cases. DESIGN: Nonrandomized, retrospective clinical study. PATIENTS AND METHODS: Nine patients (15 eyes) who underwent cataract surgery after prior PRK to correct myopia were identified. The medical records of both the laser and cataract surgery centers were reviewed. MAIN OUTCOME MEASURES: Eight different keratometric values (K values; measured or calculated) were entered into 3 different IOL calculation formulas: SRK/T, Holladay 1, and Hoffer Q. The actual biometry and IOL parameters were used to predict postoperative refraction, which was compared with the actual refractive outcome. Also, the relative underestimation of the refractive change in corneal dioptric power by keratometry after PRK was calculated. RESULTS: In 7 of 15 eyes, IOL exchange or piggybacking was performed because of hyperopia. Retrospectively, the most accurate K value for IOL calculation was found to be the pre-PRK K value corrected by the spectacle plane change in refraction. Use of the Hoffer Q formula would have avoided postoperative hyperopia in more cases than the other formulas. The mean underestimation of the change in corneal power after PRK varied from 42% to 74%, depending on the method of calculation. CONCLUSION: The predictability of IOL calculation for cataract surgery after PRK can be improved by using a corrected, refraction-derived K value instead of the measured, preoperative K value. PMID- 11934318 TI - Macular translocation with 360 degrees retinotomy for exudative age-related macular degeneration. AB - BACKGROUND: Macular rotation surgery comprises surgical extraction of choroidal neovascular membranes in age-related macular degeneration (AMD) and translocation of the foveal neural retina over adjacent retinal pigment epithelium. OBJECTIVE: To determine whether macular translocation with 360 degrees retinotomy can stabilize and/or improve visual acuity in patients with subfoveal choroidal neovascularization (CNV) secondary to AMD. DESIGN: This study consisted of a standardized surgical procedure on a series of 90 consecutive patients and follow up examinations at fixed intervals for 12 months. PARTICIPANTS: All patients in this study had experienced recent visual loss resulting from subfoveal CNV caused by AMD. Twenty-six patients had major macular subretinal hemorrhage, 39 patients had occult subfoveal CNV, and 25 patients had classic subfoveal CNV. METHODS: Macular translocation surgery was performed between 1997 and 1999. The patients were examined preoperatively and at 3, 6, and 12 months postoperatively, including visual acuity, microperimetry, angiography, and orthoptic assessment. RESULTS: Visual acuity increased by 15 or more letters in 24 patients, remained stable in 37 patients, and deteriorated by 15 or more letters in 29 patients at 12 months postoperatively. A secondary procedure was necessary in 17 patients because of severe complications; proliferative vitreoretinopathy was observed in 17 eyes, macular pucker in 5 eyes, and macular hole in 1 patient. CONCLUSION: Macular translocation is a technically demanding surgical procedure. Although the procedure has a high rate of surgical and postoperative complications, the functional and anatomical results appear to be promising for selected patients with subfoveal CNV secondary to AMD. PMID- 11934319 TI - Factors predictive of recurrence of retinal tumors, vitreous seeds, and subretinal seeds following chemoreduction for retinoblastoma. AB - OBJECTIVE: To identify the clinical features of eyes with retinoblastomas that predict the recurrence of retinal tumors, vitreous seeds, and subretinal seeds following treatment with chemoreduction. DESIGN: Prospective nonrandomized single center clinical trial. SETTING: Ocular oncology service at Wills Eye Hospital of Thomas Jefferson University (Philadelphia, Pa) in conjunction with the division of oncology at Children's Hospital of Philadelphia. PARTICIPANTS: There were 158 eyes with 364 tumors in 103 consecutive patients with retinoblastoma managed with chemoreduction between June 1994 and August 1999. INTERVENTION: All patients received treatment for retinoblastoma with 6 cycles of chemoreduction using vincristine, etoposide, and carboplatin combined with focal treatment (cryotherapy, thermotherapy, or plaque radiotherapy) for each retinal tumor. MAIN OUTCOME MEASURES: The 3 main outcome measures included recurrence of retinal tumors, recurrence of vitreous seeds, and recurrence of subretinal seeds. The clinical features at the initial examination were analyzed for their association with the main outcome measures using a series of Cox proportional hazards regressions. RESULTS: All retinal tumors, vitreous seeds, and subretinal seeds showed an initial favorable response of regression during this treatment regimen. Using Kaplan-Meier estimates, at least 1 retinal tumor recurrence per eye was found in 37% of eyes at 1 year, 51% at 3 years, and no further increase at 5 years. By multivariate analysis, the only factor predictive of retinal tumor recurrence was the presence of tumor-associated subretinal seeds at the initial examination. Of the 54 eyes that had vitreous seeds at the initial examination, vitreous seed recurrence was found in 26% of eyes at 1 year, 46% at 3 years, and 50% at 5 years. By univariate analysis, the only factor predictive of vitreous seed recurrence was the presence of tumor-associated subretinal seeds at the initial examination. Of the 71 eyes that had subretinal seeds at the initial examination, subretinal seed recurrence was detected in 53% of eyes at 1 year, 62% at 3 years, and no further increase at 5 years. By multivariate analysis, factors predictive of subretinal seed recurrence included a tumor base greater than 15 mm and a patient age of 12 months or younger at diagnosis. There were no patients who developed retinoblastoma metastasis, pinealoblastoma, or second malignant neoplasms. CONCLUSIONS: Chemoreduction combined with focal therapy is effective for selected eyes with retinoblastomas. Eyes with subretinal seeds at initial examination are at particular risk for recurrence of retinal tumor and vitreous seeds. Younger patients with large tumors are at risk for recurrence of subretinal seeds. Retinal tumor and subretinal seed recurrence seems to manifest within 3 years of follow-up. Close follow-up of all patients treated with chemoreduction is warranted. PMID- 11934320 TI - Immunophenotypic differences between uveal and cutaneous melanomas. AB - OBJECTIVE: To determine the immunophenotypic differences between uveal and cutaneous melanomas, employing standard melanoma markers as well as p75 neurotrophin receptor (p75NTR) and microphthalmia transcription factor (MITF). DESIGN: Fifteen uveal melanomas (5 spindle, 5 epithelioid, and 5 mixed uveal subtypes) were immunolabeled with a panel of antibodies that included S100, tyrosinase, melan-A, HMB-45 and HMB-50 combination, MITF, and p75NTR. The results were tabulated on the basis of intensity and pervasiveness of the labeling and compared with a prior study on cutaneous spindle and epithelioid melanomas. RESULTS: In contrast to its strong labeling of cutaneous melanomas, S100 immunolabeling of uveal melanomas was weak and variable. p75NTR, known to differentiate spindle from epithelioid melanomas of the skin, did not immunolabel uveal melanomas. HMB-45, HMB-50, tyrosinase, melan-A, and MITF immunolabeled all uveal melanomas strongly, irrespective of the histologic subtype, but not cutaneous melanomas. Microphthalmia transcription factor was especially clear in its labeling of uveal melanomas. CONCLUSIONS: Although cutaneous and uveal melanomas share many molecular markers in common, there are differences between the 2 types of melanoma. First, the level of expression of S100 differs between cutaneous and uveal melanomas. Second, while cutaneous melanomas can be further subdivided into spindle and epithelioid types based on their immunophenotype, the uveal melanomas cannot. PMID- 11934321 TI - Glaucoma in Zulus: a population-based cross-sectional survey in a rural district in South Africa. AB - OBJECTIVES: To determine the prevalence and the main types of glaucoma in a representative adult population in rural Zululand, and to describe the distribution of glaucoma-related variables in healthy subjects and those with glaucoma. DESIGN: A population-based, cross-sectional study. SETTING: Hlabisa district, Northern KwaZulu-Natal Province, South Africa. PARTICIPANTS: Resident individuals of Zulu ethnic origin, 40 years or older. MAIN OUTCOME MEASURES: Glaucoma was diagnosed by means of strict objective criteria, based on binocular indirect ophthalmoscopic optic disc appearances validated by results of disc photography and threshold visual field testing. RESULTS: From an eligible sample of 1115 subjects, 1005 (90.1%) were examined in the survey. The adjusted prevalence of glaucoma of all types was 4.5%, and primary open-angle glaucoma accounted for 2.7%. Secondary glaucoma occurred with an adjusted prevalence of 1.7%, of which the principal contributors were exfoliative and aphakic glaucoma. The prevalence of primary angle-closure glaucoma was low. Normal tension (intraocular pressure, < or =21 mm Hg) was measured in 16 (57.1%) of 28 cases of primary open-angle glaucoma. Age- and sex-adjusted prevalence of bilateral blindness was 3.2%, which was exclusively due to glaucoma in 9 (22.0%) of 41 cases. CONCLUSIONS: Primary and secondary glaucoma constitute a significant public health problem in rural Zululand. The prevalence and types of glaucoma vary among different black populations. PMID- 11934322 TI - Quality of life with visual acuity loss from diabetic retinopathy and age-related macular degeneration. AB - OBJECTIVE: To compare the quality of life in patients with visual acuity loss occurring secondary to diabetic retinopathy with visual acuity loss occurring secondary to age-related macular degeneration (ARMD). METHODS: Consecutive patients with diabetic retinopathy and ARMD were evaluated using the time trade off method of utility value analysis. Both groups were stratified according to the degree of visual acuity loss in the better-seeing eye (group 1: 20/20-20/25, group 2: 20/30-20/40, group 3: 20/50-20/100, group 4: < or =20/200). Utility values obtained from the patients, once stratified for visual acuity group, were compared with use of the t test and the Mann-Whitney U test. In addition, a 2-way analysis of variance was performed to control for potential confounding variables. RESULTS: No difference was found between the utility value means of the diabetic retinopathy (n = 333) and ARMD (n = 246) subgroups stratified according to visual acuity levels: group 1, P =.54; group 2, P =.96; group 3, P =.09; and group 4, P =.32. A 2-way analysis of variance demonstrated that, among the variables of ocular disease, sex, age, and visual acuity in the better-seeing eye, only visual acuity was significantly associated with utility values (P =.003). CONCLUSIONS: At similar levels of visual acuity loss, that associated with diabetic retinopathy causes a similar reduction in quality of life to that associated with ARMD. This information has important implications for use in cost utility analyses of ophthalmic interventions. PMID- 11934323 TI - Butterfly-shaped pattern dystrophy: a genetic, clinical, and histopathological report. AB - OBJECTIVES: To identify the disease-causing mutation in a large family segregating dominantly inherited butterfly-shaped pattern dystrophy (BPD) and to describe the microscopic pathological changes observed in a member of this family. METHODS: Seventeen individuals at risk for dominantly inherited BPD in a family were examined and blood samples obtained. Linkage analysis and mutation screening of the human retinal degeneration slow (RDS)/peripherin locus were performed. Light and electron microscopic examinations were performed on 1 postmortem eye of 1 affected individual. RESULTS: Four individuals demonstrated macular degenerative changes with diminished visual acuity, and 3 others exhibited early signs of atrophy without visual deficits. Microscopic examination of the left eye of 1 patient revealed an area of total loss of the retinal pigment epithelium (RPE) and photoreceptor cell layer with intact choriocapillaris and lipofuscin-containing cells in the subretinal space. Outside the area of RPE atrophy, the RPE was greatly distended by lipofuscin. The disease locus in this family was mapped to 6p21.2, the region of the RDS/peripherin gene. Further analysis identified a G-->A change at nucleotide position 637 of RDS/peripherin, predicting a novel Cys213Tyr substitution in all affected members of the family. CONCLUSIONS: This study describes a new RDS/peripherin mutation for BPD and provides the first combined genetic-pathological study of this condition, to our knowledge. CLINICAL RELEVANCE: Accumulation of lipofuscin in RPE is a prominent feature of several retinal disorders, including age-related macular degeneration. Further elucidation of the cellular and molecular mechanism of BPD may provide insight into pathogenesis and lead to novel treatment approaches for this and other macular degenerations. PMID- 11934324 TI - New system for fiberoptic-free bimanual vitreous surgery. AB - OBJECTIVES: To describe a new system for fiberoptic-free bimanual vitrectomy and to present the outcome of 37 eyes with preretinal membranes due to diabetic retinopathy or proliferative vitreoretinopathy that underwent surgery using this system. METHODS: The system consists of a 40-diopter aspheric field lens suspended from the operating microscope and a prismatic inverting device. The aspheric lens is placed above the cornea, and the illumination from the operating microscope creates an inverted image of the fundus, which is made erect by an inverter system. No fiberoptics are required, and both hands are free to use 2 microinstruments. MAIN OUTCOME MEASURES: The practical utility of this system and its surgical results and complications. RESULTS: The system was used successfully in all cases. Membrane dissection and hemostasis were performed without incident. An improvement in visual acuity of 2 or more lines was found in 30 of 37 eyes. Five eyes did not reveal change of 2 or more lines, and 2 eyes had a decrease in visual acuity of more than 2 lines. There was no evidence of phototoxicity. CONCLUSION: This system is very useful for bimanual vitrectomy. PMID- 11934325 TI - Lamina puncture: pars plana optic disc surgery for central retinal vein occlusion. AB - OBJECTIVE: To determine the feasibility of creating a perivascular space adjacent to the central retinal vein at the level of the lamina cribrosa as a potential method of reestablishing perfusion in central retinal vein occlusion. METHODS: Various designs for a puncture instrument, or lamina puncture lancet, were investigated in cadavers, pigs that had undergone enucleation, and in vivo rabbit eyes. RESULTS: A lancet with a sharp cutting edge on one side and an opposing blunt edge is repeatedly able to create a perivascular space with limited optic nerve fiber damage. CONCLUSIONS: Lamina puncture is technically feasible, and evaluation in carefully selected patients appears warranted. PMID- 11934326 TI - Calculating intraocular lens power after refractive corneal surgery. PMID- 11934327 TI - Clinicopathologic reports, case reports, and small case series: clinicopathologic correlation of idiopathic polypoidal choroidal vasculopathy. PMID- 11934328 TI - Clinicopathologic reports, case reports, and small case series: rapidly progressive T-cell lymphoma of the conjunctiva. PMID- 11934329 TI - Clinicopathologic reports, case reports, and small case series: congenital mydriasis, failure of accommodation, and patent ductus arteriosus. PMID- 11934330 TI - Clinicopathologic reports, case reports, and small case series: cytomegalovirus retinitis in patients with Good syndrome. PMID- 11934331 TI - Clinicopathologic reports, case reports, and small case series: unilateral, idiopathic leopard-spot lesion of the retinal pigment epithelium. PMID- 11934332 TI - Clinicopathologic reports, case reports, and small case series: retinal venous occlusion as the initial sign of tetralogy of Fallot. PMID- 11934333 TI - Candida endophthalmitis after tattooing in an asplenic patient. PMID- 11934334 TI - Peripheral curvilinear pigmentary streak in multifocal choroiditis. PMID- 11934335 TI - Incidence studies on open-angle glaucoma. PMID- 11934337 TI - Uveal melanoma in Asian Indians: a clinicopathological study. PMID- 11934338 TI - Synthetic dopamine agonists. PMID- 11934339 TI - Is pharmacological prevention of Alzheimer's a realistic goal? AB - A growing body of evidence suggests that several classes of drugs marketed for other indications may be effective in the prevention of Alzheimer's disease. Among the most promising of these are nonsteroidal anti-inflammatory agents, oestrogens (oestrogen replacement therapy) and antioxidant vitamins. Other less well-established candidates include histamine H(2) receptor antagonists (H(2) blockers) and 3-hydroxy-3-methylglutaryl coenzyme A reductase inhibitors (statins). For each of these, we discuss possible mechanisms for their postulated neuroprotective effects and review the studies suggesting their benefits in Alzheimer's disease. We conclude that nonsteroidal anti-inflammatory drugs and oestrogen replacement therapies may be effective in preventing Alzheimer's disease only if taken during the latent phase of the disease several years prior to the appearance of disturbances. Antioxidants may also prevent Alzheimer's disease, but unlike nonsteroidal anti-inflammatory drugs and oestrogen replacement therapies, they may continue to have beneficial effects even after the clinical onset of the disease. The only way to demonstrate the efficacy of these agents will be through randomised, controlled prevention trials. Such trials are currently underway but conclusive results may not be available for several years. Although intriguing, more studies on the neuroprotective effects of statins and H(2) blockers are needed before trials of these agents are initiated. Finally, there are other classes of pharmacological compounds emerging on the horizon, including folic acid, anti-beta-sheet conformational agents, secretase inhibitors and vaccines, that may soon prove to be effective for the prevention of Alzheimer's disease. PMID- 11934340 TI - Drug treatment of non-motor symptoms in Parkinson's disease. AB - Non-motor symptoms may considerably reduce parkinsonian quality of life, particularly in advanced stages of the disease. Autonomic features, such as seborrhoea, hyperhidrosis, orthostatic hypotension, excessive salivation, bladder dysfunction and GI disturbances, and neuropsychiatric symptoms, such as depression, sleep disorders, psychosis and dementia, appear in the course of Parkinson's disease. Pharmacotherapy of these non-motor symptoms complicates long term antiparkinsonian combination drug therapy due to possible drug interactions, side effects and changes in metabolism. Moreover, antiparkinsonian compounds themselves contribute to the onset of these non-motor symptoms to a considerable extent. This complicates differentiation between the disease process itself and drug-related effects, thus influencing therapeutic options, which are often limited because of comorbidity and polypharmacy. Therefore, standardised recommendations are questionable, since drug tolerability and response differ between patients. Nevertheless, this review tries to provide a survey of possible therapeutic options for the treatment of the symptoms of Parkinson's disease other the dopamine-sensitive motor features. PMID- 11934341 TI - Pregnancy and chronic headache. AB - Headache patterns in women change in relation to fluctuations in oestrogen levels. Increasing oestrogen levels in early pregnancy offer a protective effect against headache, particularly for women with migraine. However, some women continue to experience troublesome headache throughout pregnancy. Headache persisting at the end of the first trimester will usually continue without improvement for the remainder of pregnancy and should be treated. Safe and effective acute care treatment options include paracetamol, opioids and anti emetics. The use of triptans during pregnancy is controversial and not broadly recommended. Safe and effective preventive treatments include relaxation, biofeedback, beta-blockers, some antidepressants and gabapentin in early pregnancy. PMID- 11934342 TI - Nitric oxide synthase inhibitors for the treatment of chronic tension-type headache. AB - Chronic tension-type headache may be caused by prolonged painful input from pericranial myofacial tissues, for example tender points, resulting in central sensitisation (increased excitability of neurons in the central nervous system). Animal studies have shown that sensitisation of pain pathways may be caused by or associated with the activation of neuronal nitric oxide synthase and the generation of nitric oxide. Furthermore, it has been shown that nitric oxide synthase inhibitors reduce central sensitisation in animal models of persistent pain. On the basis of this information, the analgesic effect of the nitric oxide synthase inhibitor L-N(G) methyl arginine hydrochloride was investigated. This drug significantly reduced headache and myofacial factors in patients with chronic tension-type headache. These studies show that nitric oxide plays a crucial role in the pathophysiology of tension-type headache. The analgesic effect of nitric oxide synthase inhibition in patients with chronic tension-type headache is probably due to a reduction in central sensitisation at the level of the spinal dorsal horn, trigeminal nucleus or both. Furthermore, inhibition of nitric oxide synthase may become a novel principle in the future treatment of chronic headache. PMID- 11934343 TI - Combining antiplatelet and thrombolytic therapies for stroke. AB - Pharmacological therapy for acute nonhaemorrhagic stroke has become a reality over the last 5 years. Mechanistically, both thrombolytic (tissue plasminogen activator and urokinase) and antiplatelet (aspirin) monotherapy have demonstrated efficacy. However, unintended actions limit the extent of clinical improvement in each circumstance. For example, in addition to excess bleeding, tissue plasminogen activator therapy has been associated with complement activation, neuronal toxicity and laminin degradation, while aspirin may reduce nitric oxide synthase activity and cerebral blood flow. Attention is now directed toward improving the therapeutic index for each class of agents. Generally, while thrombolytic therapy is focused on developing agents with greater fibrin specificity and safety (that is, a reduction in intracranial haemorrhage rate), the development of antiplatelet agents is primarily focused on achieving greater potency. The latter is being investigated by combining agents with different mechanisms (aspirin and dipyridamole, aspirin and clopidogrel) as well as agents designed to block the glycoprotein IIb/IIIa receptor, the final common pathway for platelet aggregation. Thus, combination therapy using both thrombolytic and antiplatelet agents will further attempt to improve the therapeutic index by increasing potency and improving the safety profile. Anecdotal case studies support the merits of this approach and are consistent with the data reported for myocardial ischaemia and interventional strategies. It is anticipated that drug therapy directed at both thrombolytic and antiplatelet targets will ultimately result in a widened therapeutic window that will allow acute stroke therapy to be administrated to a much greater number of patients than is currently possible. PMID- 11934344 TI - Treatment options for renovascular hypertension. AB - Renovascular hypertension is usually caused by atherosclerotic narrowing of the origin of the renal artery and is much more common than is thought among patients with peripheral vascular disease, carotid stenosis or heart failure. Renovascular hypertension must be distinguished from renal artery stenosis. In true renovascular hypertension, the kidney takes charge of the blood pressure and will do what it takes to push blood pressure high enough to force blood through the blocked artery. This can be diagnosed with functional tests that measure glomerular filtration rate before and after blockade of the renin-angiotensin system with angiotensin converting enzyme inhibitors or antagonists of the AT(1) subtype of the angiotensin receptor. There is insufficient data on which to make evidence-based recommendations on the management of renovascular hypertension. Only two randomised trials exist of angioplasty versus medical therapy and of these the larger was severely contaminated by angioplasty among the group initially assigned to medical therapy. Only one trial exists of angiotensin converting enzyme inhibition versus alternative medical therapy. The drugs that are most effective in medical management of renovascular hypertension- angiotensin converting enzyme inhibitors and angiotensin receptor-1 blockers- tend to be avoided because of fear of a very rare complication (acute renal failure in patients with severe stenosis of both renal arteries, or the artery to a single remaining kidney). This fear is misplaced not only because it is rare (< 5% of patients with renovascular hypertension) but because it is reversible and treatable by revascularisation. Patients with renovascular hypertension should be evaluated by nuclear medicine differential glomerular filtration rate, enhanced by blockers of the renin-angiotensin system. If medical therapy is ineffective or causes severe impairment of renal function, revascularisation is required. Some experts favour surgical revascularisation because of occasional angioplasty failure and the risk of deterioration of renal function after angioplasty. PMID- 11934345 TI - Pharmacoeconomics of long-term treatment of rheumatoid arthritis. AB - Rheumatoid arthritis affects ~ 1% of the population. It is associated with pain, deformity, decreased quality of life and disability that in turn affects patients' ability to work. A variety of disease-modifying antirheumatic drugs are available to control the disease activity of rheumatoid arthritis. The goal of treatment is to improve patients' quality of life and prevent joint destruction. This paper reviews both the clinical aspects of frequently prescribed disease modifying antirheumatic drugs and the available cost-effectiveness information. Clinical evidence supports the effectiveness of methotrexate, etanercept, infliximab, gold, hydroxychloroquine, leflunomide, sulfasalazine, penicillamine, cyclosporin, azathioprine and corticosteroids. The last four of these are associated with greater toxicity and are only used if less toxic drugs are ineffective. The lack of published economic evaluations of disease-modifying antirheumatic drugs highlights the need for such studies to allow efficacious and cost-effective drugs to be used to prevent the long-term complications of uncontrolled rheumatoid arthritis. PMID- 11934346 TI - Treatment of breast cancer during pregnancy. AB - The management of breast cancer during pregnancy is one of the great clinical challenges in oncology. Patients are best served by care provided through a multidisciplinary team including surgeons, oncologists, obstetricians and genetics counsellors with experience in caring for similar women. The risks of diagnostic and therapeutic interventions can be mitigated by the consideration of known side effects of therapy on the fetus and the mother. However, because of the limited amount of clinical experience available, the potential risks to the patient, to the fetus and to the pregnancy are difficult to quantify. Treatment decisions need to be tailored carefully to the individual, respecting both her clinical circumstances and her personal preferences. PMID- 11934347 TI - The complex relationship between pregnancy and breast cancer. PMID- 11934348 TI - Rasburicase: a potent uricolytic agent. AB - Rasburicase (Fasurtec, Elitek, Sanofi-Synthelabo) is a new recombinant urate oxidase developed for the prevention and treatment of hyperuricaemia. It has a half-life of 17-21 h and produces rapid (within 4 h) and pronounced reductions in plasma uric acid concentrations. To date, rasburicase has been tested in four clinical trials conducted in patients with cancer (primarily haematological malignancies). It successfully alleviated hyperuricaemia in 98% of patients and prevented this complication in 99.6% of those who were at risk. In a stratified, randomised trial, rasburicase was more effective than allopurinol. Most patients had improved or stabilised renal function during rasburicase treatment, despite ongoing chemotherapy-induced tumour lysis. Except for occasional instances of haemolytic anaemia and methemoglobinaemia in patients with glucose-6-phosphate dehydrogenase deficiency, rasburicase was well-tolerated, with a low frequency of mild adverse events. Rasburicase is a safe and effective agent in the prevention or treatment of hyperuricaemia in cancer patients. PMID- 11934349 TI - Oxybutynin chloride: alterations in drug delivery and improved therapeutic index. AB - Oxybutynin chloride (Ditropan, Alza) is widely regarded as the most efficient antimuscarinic agent for the treatment of bladder detrusor dysfunction resulting in urinary urgency, frequency and urge incontinence. Oxybutynin metabolism occurs primarily in the proximal gastrointestinal tract and the hepatic circulation and is mediated by the cytochrome P450 3A4 isozyme. The major degradation products are desethyloxybutynin, which possesses pharmacological activity, and phenylcyclohexylglycolic acid, which is metabolically inert. A major limitation to long-term compliance with immediate-release oxybutynin remains the necessity for twice- or thrice-daily dosing regimens to provide sustained pharmacological efficacy. Side effects induced by cytochrome P450 metabolism of oxybutynin into the primary metabolite desethyloxybutynin within the gut wall substantially affect the tolerability of the compound within the individual. The oral osmotic delivery system provides unique advantages for drug delivery and substantially alters the tolerability profile of the oxybutynin chloride compound. This extended-release formulation consists of a two component core encapsulated by a semi-permeable membrane. The osmotic gradient between the surrounding environment and the inner core of the delivery system remains constant and water absorption within the capsule is controlled by the semipermeable membrane causing a controlled release of drug, which is sustained over 24 h. Herein are reviewed the various pre- and post-approval trials which have documented the overall therapeutic index of the oral osmotic oxybutynin (Ortho-McNeil Pharmaceuticals). Subsequent post-market surveillance issues are reviewed as are new developments in oxybutynin delivery. PMID- 11934350 TI - Fondaparinux versus enoxaparin for the prevention of venous thromboembolism. AB - Venous thromboembolism is a frequent, life-threatening, postoperative complication of hip-fracture and total-knee-replacement surgery. Fondaparinux is a synthetic polysaccharide that selectively binds to antithrombin, the primary endogenous regulator of blood coagulation. Low molecular weight heparins, such as enoxaparin, are less specific inhibitors of coagulation. In patients undergoing hip-fracture surgery, fondaparinux is more effective than once-daily enoxaparin as prophylaxis for venous thromboembolism. Fondaparinux (25 mg/day sc.) was also more effective than enoxaparin (30 mg sc. b.i.d.) as prophylaxis for venous thromboembolism in elective knee surgery. These differences may be explained by the fact that there is less prophylaxis cover with enoxaparin, as it has a much shorter duration of action than fondaparinux. Thus, with the present dosing regimens, fondaparinux is probably preferable to enoxaparin for the prevention of venous thromboembolism. PMID- 11934351 TI - Pre-dialysis clinic attendance improves quality of life among hemodialysis patients. AB - BACKGROUND: Although previous research has demonstrated that referral to pre dialysis clinics is associated with favourable objective outcomes, the benefit of a pre-dialysis clinic from the perspective of patient-perceived subjective outcomes, such as quality of life (QOL), is less well defined. METHODS: A retrospective incident cohort study was conducted to determine if pre-dialysis clinic attendance was a predictor of better QOL scores measured within the first six months of hemodialysis (HD) initiation. Inclusion criteria were HD initiation from January 1 1998 to January 1 2000, diagnosis of chronic renal failure, and completion of the QOL questionnaire within six months of HD initiation. Patients receiving HD for less than four weeks were excluded. An incident cohort of 120 dialysis patients was identified, including 74 patients who attended at least one pre-dialysis clinic and 46 patients who did not. QOL was measured using the SF 36 Item Health Survey. Independent variables included age, sex, diabetes, pre dialysis clinic attendance and length of attendance, history of ischemic heart disease, stroke, peripheral vascular disease, heart failure, malignancy, and chronic lung disease, residual creatinine clearance at dialysis initiation, and kt/v, albumin and hemoglobin at the time of QOL assessment. Bivariate and multivariate linear regression analyses were used to identify predictors of QOL scores. RESULTS: Multivariate analysis suggested that pre-dialysis clinic attendance was an independent predictor of higher QOL scores in four of eight health domains (physical function, p < 0.01; emotional role limitation, p = 0.01; social function, p = 0.01; and general health, p = 0.03), even after statistical adjustment for age, sex, residual renal function, kt/v, albumin, and co-morbid disease. Pre-dialysis clinic attendance was also an independent predictor of the physical component summary score (p = 0.03). CONCLUSIONS: We conclude that pre dialysis clinic attendance favourably influences patient-perceived quality of life within six months of dialysis initiation. PMID- 11934352 TI - Drug therapy and adverse drug reactions to terbutaline in obstetric patients: a prospective cohort study in hospitalized women. AB - BACKGROUND: Adverse drug reactions (ADR's) could be expected more frequently in pregnant women. This study was performed in order to identify ADR's to tocolytic drugs in hospitalised pregnant women. METHODS: A prospective cohort study was performed in two General Hospitals of the Instituto Mexicano del Seguro Social (IMSS) in Mexico City. Two hundred and seven women undergoing labor, premature labor, threatened abortion or suffering any obstetric related disease were included. Drug prescription and signs and symptoms of any potential ADR were registered daily during the hospital stay. Any potential ADR to tocolytic drugs was evaluated and classified by three of the authors using the Kramer's algorithm. RESULTS: Of the 207 patients, an ADR was positively classified in 25 cases (12.1%, CI95% 8.1 to 17.5%). All ADR's were classified as minor reactions. Grouping patients with diagnosis of threatened abortion, premature labor or under labor (n= 114), 24 ADR's were related to terbutaline, accounting for a rate of 21.1 ADR's per 100 obstetric patients. Obstetric patients suffering an ADR were older than obstetric patients without any ADR. However, the former received less drugs/day x patient-1 and had a shorter hospital stay (p < 0.05) whereas the dose of terbutaline was similar between the two groups. Terbutaline inhibited uterine motility in women with and without any ADR at a similar rate, 70 and 76% respectively (x2 = 0.07; p = 0.8). CONCLUSION: Terbutaline, used as a tocolytic drug, was related to a high frequency of minor ADRs and to a high rate of effcicacy. PMID- 11934353 TI - Lens epithelial cell apoptosis and intracellular Ca2+ increase in the presence of xanthurenic acid. AB - BACKGROUND: Xanthurenic acid is an endogenous product of tryptophan degradation by indoleamine 2,3-dioxygenase (IDO). We have previously reported that IDO is present in mammalian lenses, and xanthurenic acid is accumulated in the lenses with aging. Here, we studied the involvement of xanthurenic acid in the human lens epithelial cell physiology. METHODS: Human lens epithelial cells primary cultures were used. Control cells, and cells in the presence of xanthurenic acid grow in the dark. Western blot analysis and immunofluorescence studies were performed. RESULTS: In the presence of xanthurenic acid human lens epithelial cells undergo apoptosis-like cell death. In the control cells gelsolin stained the perinuclear region, whereas in the presence of 10 microM xanthurenic acid gelsolin is translocated to the cytoskeleton, but does not lead to cytoskeleton breakdown. In the same condition caspase-3 activation, and DNA fragmentation was observed. At low (5 to 10 microM) of xanthurenic acid concentration, the elongation of the cytoskeleton was associated with migration of mitochondria and cytochrome c release. At higher concentrations xanthurenic acid (20 microM and 40 microM) damaged mitochondria were observed in the perinuclear region, and nuclear DNA cleavage was observed. We observed an induction of calpain Lp 82 and an increase of free Ca2+ in the cells in a xanthurenic acid concentration-dependent manner. CONCLUSIONS: The results show that xanthurenic acid accumulation in human lens epithelial cells disturbs the normal cell physiology and leads to a cascade of pathological events. Xanthurenic acid induces calpain Lp82 and caspases in the cells growing in the dark and can be involved in senile cataract development. PMID- 11934354 TI - Genetically modified parthenocarpic eggplants: improved fruit productivity under both greenhouse and open field cultivation. AB - BACKGROUND: Parthenocarpy, or fruit development in the absence of fertilization, has been genetically engineered in eggplant and in other horticultural species by using the DefH9-iaaM gene. The iaaM gene codes for tryptophan monoxygenase and confers auxin synthesis, while the DefH9 controlling regions drive expression of the gene specifically in the ovules and placenta. A previous greenhouse trial for winter production of genetically engineered (GM) parthenocarpic eggplants demonstrated a significant increase (an average of 33% increase) in fruit production concomitant with a reduction in cultivation costs. RESULTS: GM parthenocarpic eggplants have been evaluated in three field trials. Two greenhouse spring trials have shown that these plants outyielded the corresponding untransformed genotypes, while a summer trial has shown that improved fruit productivity in GM eggplants can also be achieved in open field cultivation. Since the fruits were always seedless, the quality of GM eggplant fruits was improved as well. RT-PCR analysis demonstrated that the DefH9-iaaM gene is expressed during late stages of fruit development. CONCLUSIONS: The DefH9 iaaM parthenocarpic gene is a biotechnological tool that enhances the agronomic value of all eggplant genotypes tested. The main advantages of DefH9-iaaM eggplants are: i) improved fruit productivity (at least 30-35%) under both greenhouse and open field cultivation; ii) production of good quality (marketable) fruits during different types of cultivation; iii) seedless fruit with improved quality. Such advantages have been achieved without the use of either male or female sterility genes. PMID- 11934355 TI - Selection for restraint in competitive ability in spatial competition systems. AB - The absence of 'super competitors' in nature is usually attributed to organisms facing trade-offs in resource allocation. Here we identify another mechanism, dependent on indirect interactions among species and non-random spatial organization, in which selection favours restraint in competitive ability. In simple spatial models of a three-species intransitive network, indirect interactions favour slower growth and selection limits the difference in growth rate among species. The mechanism involves a trade-off between selection at the individual level, which selects for increased growth rate, and at the community level, which acts to limit growth rate to less than the maximum possible. If the difference in growth rates among species becomes too large, then the community becomes unstable and collapses to a monoculture of the slowest growing species. The mechanism requires both the intransitive network structure and self-organized spatial structure in the system. Similar behaviours arise in more complex systems of more than three species, and where there are reversals in interaction outcomes between species pairs. The work suggests that spatial self-structuring, indirect interactions and selection acting on community properties can be important in evolution. It provides a partial explanation of the high level of species coexistence and apparent restraint in interspecific interactions evident in some assemblages of sessile marine colonial organisms. PMID- 11934357 TI - Disease in endangered metapopulations: the importance of alternative hosts. AB - Conventional applications of metapopulation theory have suggested that increasing migration between patches is usually good for conservation. A recent analysis by Hess has pointed out a possible exception to this: when infectious disease is present, migration may promote disease spread and therefore increase local extinction. We extend Hess's model to discuss this problem: when infections have spilled over from more abundant alternative hosts. This is often the case for species of conservation concern, and we find that Hess's conclusions must be substantially modified. We use deterministic analytic and stochastic numerical approaches to show that movement between patches will rarely have a negative impact, even when the probability of external infection is low. PMID- 11934356 TI - A possible role for imprinted genes in inbreeding avoidance and dispersal from the natal area in mice. AB - The expression of a subset of mammalian genes is subject to parent of origin effects (POE), most of which can be explained by genomic imprinting. Analysis of mutant animals has demonstrated that a number of imprinted genes influence brain development and behaviour. Here we provide evidence for POE on olfactory related behaviour and sensitivity to maternal odour cues. This was investigated by examining the odour preference behaviour of reciprocal cross F(1) mice made by embryo transfer to genetically unrelated foster parents. We determined that both adult males and females show an avoidance of female urinary odours of their genetic maternal but not paternal origin. This was found not to be due to any previous exposure to these odours or due to self-learning, but may be related to direct effects on the olfactory system, as reciprocal F(1) males show differential sensitivity to female odour cues. Currently the most robust theory to explain the evolution of imprinting is the conflict hypothesis that focuses on maternal resource allocation to the developing foetus. Kinship considerations are also likely to be important in the selection of imprinted genes and we discuss our findings within this context, suggesting that imprinted genes act directly on the olfactory system to promote post-weaning dispersal from the natal area. PMID- 11934358 TI - Environmental stress, inbreeding, and the nature of phenotypic and genetic variance in Drosophila melanogaster. AB - Fifty-two lines of Drosophila melanogaster founded by single-pair population bottlenecks were used to study the effects of inbreeding and environmental stress on phenotypic variance, genetic variance and survivorship. Cold temperature and high density cause reduced survivorship, but these stresses do not cause repeatable changes in the phenotypic variance of most wing morphological traits. Wing area, however, does show increased phenotypic variance under both types of environmental stress. This increase is no greater in inbred than in outbred lines, showing that inbreeding does not increase the developmental effects of stress. Conversely, environmental stress does not increase the extent of inbreeding depression. Genetic variance is not correlated with environmental stress, although the amount of genetic variation varies significantly among environments and lines vary significantly in their response to environmental change. Drastic changes in the environment can cause changes in phenotypic and genetic variance, but not in a way reliably predicted by the notion of 'stress'. PMID- 11934359 TI - Sanctions and mutualism stability: why do rhizobia fix nitrogen? AB - Why do rhizobia expend resources on fixing N(2) for the benefit of their host plant, when they could use those resources for their own reproduction? We present a series of theoretical models which counter the hypotheses that N(2) fixation is favoured because it (i) increases the exudation of useful resources to related rhizobia in the nearby soil, or (ii) increases plant growth and therefore the resources available for rhizobia growth. Instead, we suggest that appreciable levels of N(2) fixation are only favoured when plants preferentially supply more resources to (or are less likely to senesce) nodules that are fixing more N(2) (termed plant sanctions). The implications for different agricultural practices and mutualism stability in general are discussed. PMID- 11934360 TI - Functional coupling of acoustic and chemical signals in the courtship behaviour of the male Drosophila melanogaster. AB - During courtship, the male Drosophila melanogaster sends signals to the female through two major sensory channels: chemical and acoustic. These signals are involved in the stimulation of the female to accept copulation. In order to determine the respective importance in the courtship of these signals, their production was controlled using genetical and surgical techniques. Males deprived of the ability to emit both signals are unable to mate, demonstrating that other (e.g. visual or tactile) signals are not sufficient to stimulate the female. If either acoustic or chemical signals are lacking, the courtship success is strongly reduced, the lack of the former having significantly more drastic effects. However, the accelerated matings of males observed with males bearing wild-type hydrocarbons compared with defective ones, whichever the modality of acoustic performance (wing vibration or playback), strongly support the role of cuticular compounds to stimulate females. We can conclude that among the possible factors involved in communication during courtship, acoustic and chemical signals may act in a synergistic way and not separately in D. melanogaster. PMID- 11934361 TI - Host range and local parasite adaptation. AB - Parasites may be expected to become locally adapted to their hosts. However, while many empirical studies have demonstrated local parasite adaptation, others have failed to demonstrate it, or have shown local parasite maladaptation. Researchers have suggested that gene flow can swamp local parasite-host dynamics and produce local adaptation only at certain geographical scales; others have argued that evolutionary lags can account for both null and maladaptive results. In this paper, we use item response theory (IRT) to test whether host range influences the likelihood of parasites locally adapting to their hosts. We collated 32 independent experiments testing for local adaptation, where parasites could be assigned as having either broad or narrow host ranges (BHR and NHR, respectively). Twenty-five tests based on BHR parasites had a significantly lower average effect size than seven NHR tests, indicating that studies based on BHR parasites are less likely to demonstrate local parasite adaptation. We argue that this may relate to evolutionary lags during diffuse coevolution of BHR parasites with their hosts, rather than differences in experimental approaches or other confounds between BHR and NHR studies. PMID- 11934362 TI - Inbreeding depression and male fitness in black grouse. AB - The male lifetime lekking performance was studied, and related to inbreeding outbreeding in a wild population of black grouse (Tetrao tetrix) in central Finland between 1989 and 1995. Inbreeding was measured as the mean heterozygosity and mean d(2) of 15 microsatellite loci. We found a significantly positive relationship between mean d(2) and lifetime copulation success (LCS), while the relationship between heterozygosity and LCS was close to significant. We also found that males that never obtained a lek territory had significantly lower mean heterozygosity than males that were observed on a territory at least during one mating season in their life. Furthermore, among males that were successful in obtaining a lek territory, LCS and mean d(2) were highest for those males that held central territories. We suggest that inbred males have a disadvantage (or outbred males have an advantage) in the competition for territories that may explain the relationships with LCS and inbreeding. Furthermore, the fact that mean d(2) was positively correlated with LCS whereas heterozygosity was not when we restricted the analysis to territorial males, suggests that mean d(2) provides more information about levels of inbreeding-outbreeding than heterozygosity alone, and potentially highlights the effects of heterosis. To our knowledge, this is the first time that measures of inbreeding and lifetime fitness have been linked in a non-isolated population. This is important in establishing that the relationships found in previous studies are not artefacts of low gene flow created by limited dispersal but a general feature of wild vertebrate populations. PMID- 11934363 TI - The effect of gravity on coral morphology. AB - Coral morphological variability reflects either genetic differences or environmentally induced phenotypic plasticity. We present two coral species that sense gravity and accordingly alter their morphology, as characterized by their slenderness (height to diameter) ratio (SR). We experimentally altered the direction (and intensity) of the gravitational resultant force acting along or perpendicular to the main body axis of coral polyps. We also manipulated light direction, in order to uncouple gravity and light effects on coral development. In the experiments, vertically growing polyps had significantly higher SR than their horizontal siblings even when grown in a centrifuge (experiencing different resultant gravitational forces in proximal and distal positions). Lowest SR was in horizontal side-illuminated polyps, and highest in vertical top-illuminated polyps. Adult colonies in situ showed the same pattern. Gravitational intensity also affected polyp growth form. However, polyp volume, dry skeleton weight and density in the various centrifuge positions, and in aquaria experiments, did not differ significantly. This reflects the coral's ability to sense altered gravity direction and intensity, and to react by changing the development pattern of their body morphology, but not the amount of skeleton deposited. PMID- 11934364 TI - Kin-recognition abilities and nepotism as a function of sociality. AB - Despite widespread interest in kin selection and nepotism, relatively little is known about the perceptual abilities of animals to recognize their relatives. Here I show that a highly nepotistic species, Belding's ground squirrels (Spermophilus beldingi), produces odours from at least two sources that correlate with relatedness ('kin labels'), and that ground squirrels can use these odours to make accurate discriminations among never before encountered ('unfamiliar') kin. Recognition odours appear to vary linearly with relatedness, rather than in an all-or-none fashion, allowing precise estimates of kinship even among distant relatives. Thus S. beldingi are able to recognize their distant kin and male kin, even though they do not treat them preferentially. I also show that a closely related species (S. lateralis) similarly produces kin labels and discriminates among kin, although it shows no evidence of kin-directed behaviour. Thus, contrary to a commonly held assumption, kin favouritism and recognition abilities can evolve independently, depending on variation in the costs and benefits of nepotism for a given species. PMID- 11934365 TI - Sexual equality in zebra finch song preference: evidence for a dissociation between song recognition and production learning. AB - Song in oscine birds is a culturally inherited mating signal and sexually dimorphic. From differences in song production learning, sex differences in song recognition learning have been inferred but rarely put to a stringent test. In zebra finches, Taeniopygia guttata, females never sing and the species has one of the greatest neuroanatomical differences in song-related brain nuclei reported for songbirds. Preference tests with sibling groups for which exposure to song had been identical during the sensitive phase for song learning in males, revealed equally strong influence of the tutor's song (here the father) on males' and females' adult song preferences. Both sexes significantly preferred the father's over unfamiliar song when having free control over exposure to playbacks via an operant task. The sibling comparisons suggest that this preference developed independently of the song's absolute quality: variation between siblings was as great as between nests. The results show that early exposure has an equally strong influence on males' and females' song preferences despite the sexual asymmetry in song production learning. This suggests that the trajectory for song recognition learning is independent of the one for song production learning. PMID- 11934366 TI - Craniotopic updating of visual space across saccades in the human posterior parietal cortex. AB - The neural mechanisms underlying the craniotopic updating of visual space across saccadic eye movements are poorly understood. Previous single-unit recording studies in primates and clinical studies in brain-damaged patients have shown that the posterior parietal cortex (PPC) has a key role in this process. In the present study, we used single-pulse transcranial magnetic stimulation (TMS) to disrupt the processing within the PPC during a task that requires craniotopic updating: double saccades. In this task, two targets are presented in quick succession and the subject is required to make a saccade to each location as accurately as possible. We show here that TMS delivered to the PPC just prior to the second saccade effectively disrupts the craniotopic coding normally observed in this task. This causes subjects to revert to saccades more consistent with a representation of the targets based on their positions relative to one another. By contrast, stimulation at earlier times between the two saccades did not disrupt performance. These results suggest that extraretinal information generated during the first perisaccadic period is not put into functional use until just prior to the second saccade. PMID- 11934367 TI - The coevolution of warning signals. AB - It has long been recognized that defended prey tend to be conspicuous. Current theories suggest that the association ('aposematism') has arisen because predators more readily learn to avoid attacking defended phenotypes when they are conspicuous. In this paper, I consider why such psychology has evolved. In particular, I argue that aposematism may have evolved not because of an independent and pre-existing receiver bias, but because the conspicuousness of a prey item provides a reliable indicator of its likelihood of being defended. To develop my case I consider how warning signals might coevolve in a system containing a number of predators, whose foraging behaviour is also subject to selection. In these cases, models readily show that the greater the conspicuousness of a novel prey item, the more likely that it has been encountered by other predators and survived. As a consequence, naive predators should be less likely to attack highly conspicuous novel prey on encounter, or at least more inclined to attack them cautiously. This adaptive predator behaviour will greatly facilitate the spread of aposematic phenotypes from extreme rarity, which in turn will enhance selection for forms of predator behaviour under which aposematism will coevolve even more readily. PMID- 11934368 TI - Variations in adult body mass in roe deer: the effects of population density at birth and of habitat quality. AB - Body mass is a key determinant of fitness components in many organisms, and adult mass varies considerably among individuals within populations. These variations have several causes, involve temporal and spatial factors, and are not yet well understood. We use long-term data from 20 roe deer cohorts (1977-96) in a 2600 ha study area (Chize, western France) with two habitats contrasting in quality (rich oak forest in the North versus poor beech forest in the South) to analyse the effects of both cohort and habitat quality on adult mass (i.e. median body mass between 4 and 10 years of age) of roe deer (Capreolus capreolus). Cohort strongly influenced the adult body mass of roe deer in both sexes: males born in 1994 were 5.2 kg heavier when aged between 4 and 10 years old than males born in 1986, while females born in 1995 were 4.7 kg heavier between 4 and 10 years old than females born in 1982. For a given cohort, adult males were, on average, 0.9 kg heavier in the rich oak forest than in the poor beech forest. A similar trend occurred for adult females (0.5 kg heavier in the oak forest). The effects of cohort and habitat were additive and accounted for ca. 40% of the variation observed in the adult mass of roe deer at Chize (males: 41.2%; females: 40.2%). Population density during the spring of the birth accounted for about 35% of cohort variation, whereas rainfall in May-June had no effect. Such delayed effects of density at birth on adult body mass probably affect population dynamics, and might constitute a mechanism by which delayed density-dependence occurs in ungulate populations. PMID- 11934369 TI - Inbreeding and parasite sex ratios. AB - The breeding system of parasitic protozoa affects the evolution of drug resistance and virulence, and is relevant to disease diagnosis and the development of chemo- and immunotherapy. A major group of protozoan parasites, the phylum Apicomplexa, that includes the aetiological agents of malaria, toxoplasmosis and coccidiosis, all have dimorphic sexual stages. The sex ratio (proportion of males produced by parasites) is predicted to depend upon the inbreeding rate, and it has been suggested that sex-ratio data offer a relatively cheap and easy method for indirectly estimating inbreeding rates. Here, we exploit a new theoretical machinery to show that there are generally valid relationships between f, Wright's coefficient of inbreeding, and sex ratio, z(*), the generality being with respect to population structure. To focus the discussion, we concentrate on malaria and show that the previously derived result, f = 1 - 2z(*), does not depend on the artificial assumptions about population structure that were previously made. Not only does this justify the use of sex ratio as an indirect measure of f, but also we argue that it may actually be preferable to measure f by measuring sex ratios, rather than by measuring departures from Hardy-Weinberg genotypic proportions both in malaria and parasites more generally. PMID- 11934370 TI - Using financial incentives to promote teamwork in health care. PMID- 11934371 TI - Health needs assessment and needs-led health service change: a survey of projects involving public health doctors. AB - OBJECTIVES: To describe the criteria that define an effective health needs assessment and to explore which factors are important for the delivery of effective health needs assessment in the English National Health Service (NHS). METHODS: A postal questionnaire to all public health doctors in an English health region to describe health needs assessment activity, including initiating factors, methods and outcomes. This was followed by semi-structured interviews with public health professionals and others involved in 10 purposively selected needs assessments. RESULTS: A response rate of 62% identified a total of 102 health needs assessments undertaken between 1993 and 1998. A number of themes emerged as being important in the impact of health needs assessments on policy and planning. These included careful design, methodological rigour, decisive leadership, good communication, involvement and ownership of the work from relevant stakeholders, support from senior decision-makers, appreciation of the political dynamics and engagement with local priorities, availability of resources and, finally, an element of chance. These themes can be categorised broadly into contextual factors and quality or robustness of the work. Our findings suggest that, although methodological and analytical quality are necessary characteristics of effective health needs assessment, they are not sufficient without a favourable political environment. CONCLUSION: There appear to be conditions under which needs assessment is more likely to be effective in terms of its influence on policy and planning. However, it is clear that needs assessment does not occupy a central position in health service decision-making, remaining vulnerable to a range of factors over which those responsible for its conduct have little or no control. PMID- 11934372 TI - The test-retest reliability of the modified Patient Generated Index. AB - OBJECTIVES: The purpose of this study was to determine the test-retest reliability of the recently modified Patient Generated Index (PGI) when used as a self-completed postal questionnaire by older people with arthritis. METHODS: Two sets of postal questionnaires were used, sent two weeks apart, followed by interviews with a theoretically chosen subsample of respondents to both. Reliability for respondents whose health had not changed in the intervening two weeks was assessed using intraclass correlation coefficients. Data from interviews were used to develop theories to select subgroups for analysis. RESULTS: One thousand one hundred and twenty-nine people over 65 years old with arthritis were surveyed and 53 were selected for interview. The response rates to the two surveys were 78% and 83%, respectively. Complete data for the test-retest analysis were available from 236 (29.2%) respondents. The intraclass correlation coefficient (r(i)) was 0.55 [95% confidence interval (CI) 0.43 to 0.64, P < 0.0001]. The most optimistic range for the limits of agreement was 64.66 around the difference between two total PGI scores. The intraclass correlation coefficient decreased considerably if there had been more than six changes to the areas chosen for inclusion on the second occasion (r(i) = 0.37, 95% CI 0.01 to 0.65, P < 0.05). Interview data showed that some respondents incorrectly interpreted the scoring instructions, often inconsistently. When their data were excluded, the intraclass correlation coefficient increased to 0.67 (95% CI 0.56 to 0.76, P < 0.0001). CONCLUSIONS: The results indicate that the modified PGI may be sufficiently reliable for within-group comparison of elderly people with arthritis, but not for assessing individual patients. However, it also elicits individual patient concerns and priorities about disease that other questionnaire based instruments may not identify. PMID- 11934373 TI - The effects of case-based pricing on length of stay for common surgical procedures. AB - OBJECTIVES: Finland's state subsidy reform of 1993 encouraged hospital districts to adopt case-based pricing of their services. The revenues of most hospitals have since depended on the volume and price of services produced. Information on the incentive effects of this method of funding is needed to further develop pricing systems in public hospitals. This study focuses on identifying the effects of case-based prices on length of stay (LOS) for lumbar discectomies and hip and knee replacements. METHOD: The effects of case-based pricing were estimated using count data models. Data from 1994-1998 on 17,767 lumbar disc patients and 19,039 hip and 15,174 knee replacement patients treated in public hospitals were used. Patient characteristics and hospital and hospital district characteristics, as well as accessibility of care and the time trend were taken into account in the modelling. RESULTS: Use of case-based pricing shortened the LOS by 0.49 days for hip replacements and 0.55 days for knee replacements, but did not shorten LOS for lumbar discectomies. CONCLUSIONS: The use of case-based prices would make possible about 400 extra knee and hip replacements yearly, resources (e.g. orthopaedists) permitting. The effects of hospital pricing reform on LOS in Finland compared with countries such as the USA and Sweden have been relatively modest. Overall, measures to reduce differences in clinical practice would probably have a greater impact on LOS or the number of cases treated than case-based pricing. PMID- 11934374 TI - Updating evidence-based clinical guidelines. AB - OBJECTIVES: To report on the updating of two evidence-based guidelines. METHODS: Development of multi-disciplinary evidence-based guidelines within the North of England Evidence Based Guideline Development Programme. RESULTS: The guideline updating groups were multi-disciplinary although the balance of disciplines was different from the original development groups; the convening of the updating groups allowed refinement of both the skills within the groups and the overall size of the groups. As both groups were starting with a previous version of the guideline, defining the scope of the guideline was more straightforward and faster than when developing the guideline de novo. The process of evidence identification for both guidelines was again by systematic review. Updating the guidelines influenced recommendations in two ways: new evidence required new recommendations; and supplementary evidence in an existing area allowed refinement of recommendations. Overall savings within the guideline updating process (compared with initial development) were modest. The major costs of both the initial guideline development and the updating process were staff costs, which were identical for the two processes. CONCLUSIONS: There is little other experience to set alongside this paper. There is a need for further sharing of experiences and the development of explicit methods for informing decisions on when and how to update evidence-based guidelines. PMID- 11934375 TI - The resurrection of hospital mortality statistics in England. AB - Mortality statistics for English hospitals were published by the Labour government in 1998, partly in response to the tragedy at the Bristol Royal Infirmary involving the deaths or serious injury of babies and children who had had unsuccessful heart operations. Despite being presented as an important innovation, this publication policy had a number of precedents, most notably the data produced as a result of Florence Nightingale's efforts in the 1860s and the clinical indicators developed by the Scottish Office in the early 1990s. In addition, league tables of death rates for English hospitals were available from 1992 to 1996, although there was widespread ignorance of their existence. This paper examines each of these precedents before discussing events that weakened public trust in the medical profession's ability to regulate itself and led to the Labour government's decision to resurrect the publication of hospital mortality data. It is suggested that these performance indicators may be largely symbolic of the government's broader commitment to performance assessment, but it is also recognised that, if coupled with appropriate incentives, public disclosure of mortality data may foster genuine performance improvements. PMID- 11934376 TI - Strategies to contain the emergence of antimicrobial resistance: a systematic review of effectiveness and cost-effectiveness. AB - OBJECTIVES: To conduct a systematic review of the literature to describe and critically appraise studies reporting on the cost and/or effectiveness of interventions proposed to control the emergence of antimicrobial resistance (AMR). METHODS: The search for relevant studies encompassed consultation with world experts in AMR, and electronic bibliographic database search of: Medline (1960-2000); ISI (1981-2000); EMBASE (1988-2000); Grey Literature (1999-2000); Database of Reviews of Effectiveness (DARE) and the NHS Health Economic Evaluation Database (HEED) at York University's Centre for Reviews and Dissemination (CRD) (numerous years); OPAC (1975-2000); and the Cochrane Library Online (1990-2000). Only studies that concerned the effectiveness or cost effectiveness of measures specifically designed to contain the emergence of AMR were reviewed. Standardised data extraction sheets, based on existing checklists for effectiveness and cost-effectiveness, were used to assess the validity of each study using the 'risk of bias criteria' suggested in the Cochrane Handbook. Only studies categorised as being at low or moderate risk of bias were reported fully. The reliability of the data review process was monitored by comparison of several, random, independent assessments by all authors. The mix of study methods (i.e. including studies based on non-randomised controlled trials) meant that formal meta-analysis was not possible, and thus a qualitative review was performed. RESULTS: In total, 43 studies were reviewed, with 21 classed as being at moderate or low risk of bias and therefore reported in the paper. These studies covered policies on: restricting the use of antimicrobials (five studies, suggesting that restriction policies can alter prescriber behaviour, although with limited evidence of subsequent effect on AMR); prescriber education, feedback and use of guidelines (six studies, with no clear conclusion); combination therapies (seven studies, showing the potential to lower drug specific resistance, although for an indeterminate time period); vaccination (three studies showing cost/effectiveness). Most of these studies were: from the developed world, principally the USA; hospital-based, with few community level interventions; and concerned with effectiveness, not cost-effectiveness. CONCLUSIONS: Overall, there is an absence of good evidence concerning what is effective, and especially cost-effective, in reducing the emergence of AMR. However, in addition to more research concerning these forms of intervention, the paper highlights four specific areas for further investigation: validating intermediate or surrogate outcome measures to enable better use to be made of the literature on intermediate measures; development and evaluation of 'macro' strategies; research into specific aspects of AMR in developing countries; and empirical and methodological research concerning the economic evaluation of interventions. PMID- 11934378 TI - The demise of linearity in managing health services: a call for post normal health care. AB - The emerging science of complex adaptive systems offers a complementary perspective on organisational analysis and is already finding an application within health care. The emphasis moves away from the features of normal science (analysis, prediction and control) to focus instead on the configuration of relationships among the system's components and an understanding of what creates patterns of order and behaviour among them. The important features are connectivity, recursive feedback, diversity and the existence of self-ordering rules that give systems the capacity to emerge to new patterns of order. This paper describes some of these complexity insights and their application to health care delivery and development. PMID- 11934381 TI - Atypical presentation of the Prader-Willi syndrome. Mosaic trisomy 15? AB - We report a female with Prader-Willi syndrome and hemihypertrophy. We discuss the possibility of an undetected mosaicism for trisomy 15 explaining this unusual feature. PMID- 11934382 TI - Reciprocal translocation between Y chromosome long arm euchromatin and the short arm of chromosome 1. AB - A case with an apparently balanced reciprocal translocation between the long arm of the Y chromosome and the short arm of chromosome 1 t(Y;1)(q11.2;p34.3) is described. The translocation was found in a phenotypically normal male ascertained by infertility and presenting for intra-cytoplasmatic sperm injection treatment. Histological examination of testicular biopsies revealed spermatogenic failure. Chromosome painting with probes for chromosome 1 and for the euchromatic part of the Y chromsome confirmed the translocation of euchromatic Y chromosomal material onto the short arm of chromosome 1 and of a substantial part of the short arm of chromosome 1 onto the Y chromosome. Among the Y/autosome translocations, the rearrangements involving long arm euchromatin of the Y chromosome are relatively rare and mostly associated with infertility. Microdeletion screening at the azoospermia locus revealed no deletions, suggesting another mechanism causing infertility in this translocation carrier. PMID- 11934383 TI - Distal trisomy of 10q. Report of a new case of duplication 10q25.2-25.3-->qter defined by FISH. AB - In the present work, we report on a 2.5-year-old male patient with typical clinical features of partial trisomy of the distal third of chromosome 10 long arm. The karyotype was: 46,XY, dir dup(10)(q25.2-25.3-->qter). The identification of the duplicated segment was carried out by the fluorescence in situ hybridization technique using region-specific probes. The proband's phenotype is compared with previously reported cases. PMID- 11934384 TI - Familial (11;21)(p13;q22)pat balanced reciprocal translocation in a female child with regression of milestones. AB - The role of balanced translocations in the human morphogenesis is difficult to interpret. A balanced reciprocal translocation (BRT) was observed in a female child referred with a history of regression of milestones. The cytogenetic findings by GTG-banding and fluorescence in situ hybridization revealed a BRT involving chromosomes 11p and 21q, i.e. 46,XX, t(11;21)(p13;q22). The father was found to be a carrier of the same BRT. This is the first report of reciprocal translocation involving 11p and 21q. The possible reasons for the manifestation of clinical features in the proband due to inherited BRT are discussed. PMID- 11934385 TI - del(X)(p22.1)/r(X)(p22.1q28) Dynamic mosaicism in a Turner syndrome patient. AB - We report on a 16-year-old patient with Turner syndrome who presented a mos 46,X,del(X)(p22.1)[35]/45,X [19]/46,X,r(X)(p22.1q28)[6]GTG-band karyotype. The R banding showed that the abnormal X-chromosome was inactive in all 61 cells analyzed. Fluorescence in situ hybridization with a Xp/Yp subtelomeric probe revealed that both abnormal chromosomes lacked the complementary sequences, a fact consistent with a terminal deletion. Besides, the molecular analysis of the human androgen receptor gene showed that the rearranged chromosome was paternal in origin. Since the deleted and the ring chromosomes had the same size and banding pattern, and because the former was the predominant cell line, it was inferred that the Xp- formed a ring in some cells apparently without further loss of genetic material. However, the reverse sequence and even a simultaneous origin due to a complex intrachromosomal exchange are also conceivable. The mild Turner syndrome phenotype is explained by the mosaicism and by the size of the deleted segment. PMID- 11934386 TI - Evaluation and evolution during time of prenatal diagnosis of congenital heart diseases by routine fetal ultrasonographic examination. AB - The objectives of this study were to evaluate routine prenatal diagnosis of congenital heart diseases (CHD) by fetal ultrasound examination in a well-defined population during the period 1994-1999 and to compare these results with the results from 1979 to 1993. This study included 80,076 consecutive pregnancies of known outcome from 1994 to 1999. CHD were classified as isolated or associated when at least one other major extra-cardiac malformation was present. Only 137 out of 688 malformed fetuses with CHD without chromosomal anomalies were detected (19.9%). The sensitivity of detection varied from 61.9% for malformations such as isolated hypoplastic left heart and single ventricle, to around 7-19% for atrial and ventricular septal defects. Prenatal detection rate of CHD was 11.4% for isolated cases, and 40.2% for multiple malformed with CHD. The gestational age at discovery varied from 16 to 36 weeks. There is no upper limit for termination of pregnancies in our country; 12.3% of all pregnancies were terminated after prenatal diagnosis. However, 62% of the pregnancies with a CHD detected prenatally were terminated. The detection rate of CHD increased during time from 9.2% during the period 1979-1988 to 13.7% during the period 1990-1993 and to 19.1% during the period 1994-1999. Our study shows large variation in the prenatal detection rate of CHD. Prenatal diagnosis of CHD is significantly higher when associated malformations are present. Cardiac defects affecting the size of the ventricles have the highest detection rate. Gestational age at discovery was 20-24 weeks for the majority of associated cardiac defects. The prenatal detection rate of CHD increased during time from 1979 to 1999. PMID- 11934388 TI - Rhabdomyosarcoma: cytogenetics of five cases using fine-needle aspiration samples and review of the literature. AB - Cytogenetic analyses of fine-needle aspiration samples were performed on five cases of which three were alveolar rhabdomyosarcomas (RMS), one was embryonal RMS and one was RMS of mixed alveolar and embryonal histology. Three cases of alveolar RMS and one case of embryonal RMS showed t(2;13). A del(1)(p11) in a mixed alveolar and embryonal RMS was observed without the presence of t(2;13). add(17)(q25) was present in one of the alveolar RMS along with a t(2;13). Modal number of chromosome in the five cases ranged from hyperdiploid to hypertetraploid. Clinical, cytological, histopathological and cytogenetic findings are correlated. The role of additional abnormalities is discussed with a review of appropriate literature. PMID- 11934387 TI - Cytogenetic analysis in 139 Tunisian patients with de novo acute myeloid leukemia. AB - This paper presents the results of a cytogenetic analysis in 139 Tunisian patients with de novo acute myeloid leukemia (AML), including 27 children aged 1 15 years and 112 adults. Mean age was 32 (range 1-75) and the M/F ratio was 1.43. Of our patients, 45% had apparently normal karyotypes. Acquired chromosome aberrations were found in 77 (55% ) patients. t(8;21) was identified in 27 patients (19%); t(15;17) in 13 patients (9%); deletion 7q or monosomy 7 in seven patients (5%); +8 in seven patients (5%); abnormal 16 in four patients (3%); 11q23 rearrangements in two patients (2%) and del(5q), in one patient (1%). The remaining 16 patients had miscellaneous clonal abnormalities. Specific translocations associated with the FAB type were found: t(8;21) with AML2 and t(15;17) with AML3. We concluded that our study in a Tunisian population confirmed the relation between some specific abnormalities and the FAB classification. We found a higher incidence for t(8;21) than usually described. PMID- 11934389 TI - Linkage disequilibrium and founder effect analysis of the NF1 gene in French Canadians from the Quebec population. AB - We genotyped 19 neurofibromatosis type 1 (NF1) families from French Canadians of the Quebec population with four intragenic microsatellites (IVS26-2.3, IVS27AC28.4, IVS27AC33.1, and IVS38GT53.0). Linkage analysis of the four microsatellite markers among the 19 NF1 families indicates that the four microsatellites are strongly linked with NF1 disease (LOD = 2.76-3.64). The four markers are associated (P = 0-0.077) except marker pair IVS26-2.3/IVS27AC33.1 (P = 0.18 or 0.17). However, perhaps due to the high mutation rate of the NF1 gene, no founder effect for NF1 was detected in the Quebec French Canadians. PMID- 11934390 TI - HLA polymorphism in type 1 diabetes Tunisians. AB - Several studies of the association between HLA and type 1 diabetes have been carried out revealing differences between ethnic groups. Our study, as part of the studies that should be performed about this association in the rest of the word, aims at elucidating the HLA DRB1, DQB1 polymorphism in Tunisian type 1 diabetes. This study includes 43 unrelated type 1 diabetes patients, and their mean age at onset is less than 15 years. Analysis of the frequency of alleles and haplotypes in these subjects, compared to a reference group (n = 101) led to the following results. 1) The Tunisian insulin-dependent diabetics present similarities as well as differences with other ethnic groups (Caucasians, North Africans). 2) The haplotype DRB1*04 DQ*0302 and DRB1*03 DQB1*0201 is positively associated to type 1 diabetes. 3) The heterozygotic genotype DRB1*04 DQB1*0302 / DRB1*03 DQB1*0201 is strongly associated to type 1 diabetes. 4) The haplotypes DRB1*01501 DQB1*0602 and DRB1*11 DQB1*0301 proved to be protective. In addition, the study of the subtypes DRB1*04 showed that alleles DRB1*0405 predispose to type 1 diabetes, whereas the allele DRB1*0403, which is in linkage disequilibrium with the DQB1*0402 in the Tunisian population, has a protective effect. PMID- 11934392 TI - The essential role of a free sulfhydryl group in blocking the cholesteryl site of cholesteryl ester transfer protein (CETP). AB - Cholesteryl ester transfer protein (CETP) has at least one unpaired sulfhydryl residue, which we have shown previously to be in or near the active site region. We investigated the location of this unpaired cysteine residue(s) of CETP using chemical modification with fluorescent sulfhydryl-specific reagents, limited proteolysis, and amino acid/sequence analysis. The kinetics of labeling CETP by either 2-(4'-maleimidylanilino)-naphthalene-6-sulfonic acid (MIANS) or acrylodan were followed by observing the increase in fluorescence of the bound probes. Labeling was inhibited strongly by preincubation of the CETP with either PNU-617, a competitive inhibitor of cholesteryl ester (CE) transport, and TP2 antibody. In addition, the transfer activities of the substrate CE by the modified CETP's were also inhibited but not competitively. Finally, preincubation of the native protein with N-ethylmaleimide (NEM) resulted in inhibition of activity that was dependent upon the time of exposure of the protein to the alkylating agent. These results provide further evidence that there is a cysteine residue in the active site region of CETP and ligands that either react or bind to this residue produce steric hindrance to CE transfer activity. Finally, although not conclusive, results of the protein chemistry experiments with the modified CETP suggest that the cysteine residue at position 333 is unpaired. PMID- 11934393 TI - Phase separation of cholesterol and the interaction of ethanol with phosphatidylserine-cholesterol bilayer membranes. AB - Thermotropic and structural effects of ethanol on phosphatidylserine (PS) membranes containing up to 0.4 mol fraction cholesterol were investigated by differential scanning calorimetry, X-ray diffraction and fluorescence spectroscopy. It was found that in the presence of cholesterol, 10% (v/v) added ethanol depresses the melting temperature of the phospholipid by approximately 2 degrees C, similar to what was observed in the absence of cholesterol. Below the melting temperature the progressive disordering effect of added cholesterol is weakly enhanced by the presence of ethanol. In the liquid crystalline state, the marked decrease in the thickness of the bilayer which ethanol causes in the absence of cholesterol (Chem. Phys. Lipids 92 (1998) 127), is also observed in its presence. We conclude that, in contrast to what has been observed for zwitterionic phospholipids, high concentrations of cholesterol do not diminish the interaction of ethanol with PS membranes. With addition of 10% (v/v) ethanol, crystalline cholesterol diffraction, an indication of phase separation of the sterol, appears at mol fraction cholesterol 0.34, as compared to 0.3 in the absence of ethanol (Chem. Phys. Lipids 92 (1998) 71). PMID- 11934394 TI - Ganglioside GM(1) biphasically regulates the activity of protein kinase C by the effects on the structure of the lipid bilayer. AB - Addition of a small amount of ganglioside GM(1) to phosphatidylserine (PS) liposomes, a gradual increase of protein kinase C (PKC) activity was recorded up to about 2 mol% GM(1) where the maximal enzyme activity was obtained. Then the activity of PKC began to decline and even turned to be inhibited with the further increase of GM(1) content. It was also indicated that GM(1)/PS binary liposomes had the highest membrane fluidity and very low spatial density of lipid headgroups which was demonstrated in the MC-540 studies due to the interposition of GM(1) when the liposomes contained about 2 mol% GM(1). Besides, the liposomes containing about 2 mol% GM(1) provided a more hydrophobic environment for PKC than the liposomes containing less or more GM(1) which was indicated in the Acrylodan experiments. These factors commonly induced PKC to be stimulated maximally. Whether at the lower or higher GM(1) content, the membrane structure was not the most suitable to support the activity of PKC, which declined as a consequence. PMID- 11934395 TI - New modified single chained glycolipids. Part 1: synthesis of deoxy and partially O-methylated glycolipids with or without a sulfur containing spacer. AB - A way to synthesize neoglycolipids with high yields and anomeric purity is described. Starting point of the synthesis strategy is the glycosylation of allyl alcohol with definite steric orientation. Introduction of the hydrophobic moiety was achieved by photoaddition of n-hexadecanethiol and 3-mercaptopropionic acid followed by amidation with n-hexadecylamine, respectively. In order to investigate the influence of different carbohydrate headgroups in the physicochemical behavior of the general glycolipid, especially the orientation of the alkyl chain, a range of neoglycolipids was synthesized. Beside the differences in the configuration between unfunctionalized glycopyranoses like D glucose, D-galactose and D-mannose, a number of deoxy and partially O-methylated sugar derivatives was prepared. The divergences concerning the different carbohydrate headgroups and the hydrophobic moiety, respectively, can be compared to relatively simple structured glycolipids with hexadecyl residue and without spacer function. PMID- 11934396 TI - Effect of urea, dimethylurea, and tetramethylurea on the phase behavior of dioleoylphosphatidylethanolamine. AB - The phase behavior of dioleoylphosphatidylethanolamine in aqueous solutions of urea, N,N'-dimethylurea (DMU), and N,N,N',N'-tetramethylurea (TMU) has been characterized by synchrotron X-ray diffraction and differential scanning calorimetry. All three solutes stabilize the lamellar liquid-crystalline phase at the expense of lamellar-gel phase and inverted hexagonal phase of the phospholipid when present in concentrations up to 3 M. X-ray diffraction data demonstrated that the repeat spacing of DOPE increased with increasing urea concentration, but decreased as the DMU and TMU concentrations increased. The repeat spacing of DOPE in the liquid-crystal phase dispersed in the three solutes is d(urea)>d(DMU)>d(TMU). The molecular mechanisms underlying these observations are discussed in terms of either membrane Hofmeister effect, where urea acts as a water structure breaker, or a direct insertion effect of the amphiphilic DMU and TMU molecules into the lipid head groups in the interfacial region of the phospholipid bilayer. PMID- 11934397 TI - Effect of cholesterol and surfactant protein B on the viscosity of phospholipid mixtures. AB - Low viscosity of the surface of alveolar fluid is mandatory for undisturbed surfactant function. Based on the known reduction of the viscosity of surfactant like phospholipid (PL-) mixtures by plasmalogens, the effect of cholesterol and surfactant protein (SP-) B on surface viscosity of these lipid mixtures has been studied. Surface viscosity at the corresponding surface tension was measured with the oscillating drop surfactometer. We found that the viscosity was lowest in cholesterol-, followed by plasmalogen- and SP-B containing samples. Addition of SP-B to a plasmalogen-containing PL-mixture caused a further decrease in viscosity. However, in cholesterol containing mixtures, addition of SP-B led to a significant increase in viscosity, and the effect was reversed by further addition of plasmalogens. We conclude that SP-B, plasmalogens and cholesterol all affect the surface viscosity, thus synergistically regulate monolayer stability. This suggests that they are all needed in vivo for fine tuning of surface properties of pulmonary surfactant. PMID- 11934398 TI - The influence of structure and redox state of prenylquinones on thermotropic phase behaviour of phospholipids in model membranes. AB - Our study was aimed to investigate the significance of the isoprenoid side chain size as well as redox state of the quinone ring for interaction of two main classes of prenylquinones: plastoquinones (PQ) and ubiquinones (UQ) with lipid bilayers. By use of differential scanning calorimetry (DSC) we have followed the thermotropic behaviour of multilamellar vesicles prepared from dipalmitoylphosphatidylcholine (DPPC) upon incorporation of increasing amount (1.3-12 mol%) of quinone (quinol) molecules. Our studies reveal that as the side chain is shorter (from 9 to 2 isoprenoid units) the height of the calorimetric profiles is reduced and the temperature of the main transition of DPPC (T(m)) decreases (T(m)=39.4 degrees C for a sample with 12 mol% of PQ-2), and then increases up to 39.8 degrees C for PQ-1. For the samples containing quinols the effect is more pronounced even at lower concentration. The greater influence of the added prenylquinones on the pretransition demonstrates a stronger distortion of the DPPC packing in the gel state. It seems that this is the isoprenoid side chain length rather than the redox state of prenylquinones that determines their effectiveness in perturbation of thermotropic properties of lipid bilayer. PMID- 11934399 TI - Synthesis of a novel fluorescent ceramide analogue and its use in the characterization of recombinant ceramidase from Pseudomonas aeruginosa PA01. AB - Ceramidase (CDase) hydrolyses the N-acyl linkage of the sphingolipid ceramide. We synthesized the non-fluorescent ceramide analogue (4E,2S,3R)-2-N-(10 pyrenedecanoyl)-1,3,17-trihydroxy-17-(3,5-dinitrobenzoyl)-4-heptadecene (10) that becomes fluorescent upon hydrolysis of its N-acyl bond. This novel substrate was used to study several kinetic aspects of the recombinant CDase from the pathogenic bacterium Pseudomonas aeruginosa PA01. Maximum CDase activity was observed above 1.5 microM substrate, with an apparent K(m) of 0.5+/-0.1 microM and a turnover of 5.5 min(-1). CDase activity depends on divalent cations without a strong specificity. CDase is inhibited by sphingosine and by several sphingosine analogues. The lack of inhibition by several mammalian CDase inhibitors such as D-erythro-MAPP, L-erythro-MAPP or N-oleoylethanolamine points to a novel active site and/or substrate binding region. The CDase assay described here offers the opportunity to develop and screen for specific bacterial CDase inhibitors of pharmaceutical interest. PMID- 11934400 TI - Iron-catalyzed reaction products of alpha-tocopherol with 1-palmitoyl-2-linoleoyl 3-sn-phosphatidylcholine (13S)-hydroperoxide. AB - Alpha-tocopherol was reacted with 1-palmitoyl-2-[(9Z,11E)-(S)-13-hydroperoxy-9,11 octadecadienoyl]-3-sn-phosphatidylcholine (13-PLPC-OOH) in the presence of a lipid-soluble iron chelate, Fe(III) acetylacetonate, in methanol at 37 degrees C. The reaction product was isolated and identified as a mixture of 1-palmitoyl-2 [(10E)-(12S,13S)-9-(8a-dioxy-alpha-tocopherone)-12,13-epoxy-10-octadecenoyl]-3-sn phosphatidylcholine and 1-palmitoyl-2-[(9Z)-(12S,13S)-11-(8a-dioxy-alpha tocopherone)-12,13-epoxy-9-octadecenoyl]-3-sn-phosphatidylcholine (TOO epoxyPLPC), in which the 12,13-epoxyperoxyl radicals derived from 13-PLPC-OOH attacked the 8a-position of the alpha-tocopheroxyl radical. The iron and ascorbate-catalyzed reaction of 13-PLPC-OOH with alpha-tocopherol in phosphatidylcholine (PC) liposomes was assessed by measuring the reaction products of alpha-tocopherol. When 13-PLPC-OOH and alpha-tocopherol were added in saturated dimyristoyl-PC liposomes, the products were TOO-epoxyPLPC, alpha tocopherylquinone, and epoxy-alpha-tocopherylquinones. In 1-palmitoyl-2-linoleoyl PC (PLPC) liposomes, alpha-tocopherol could react with both the 13-PLPC-OOH derived 12,13-epoxyperoxyl radicals and the PLPC-derived peroxyl radicals and formed the addition products together with alpha-tocopherylquinone and epoxy alpha-tocopherylquinones. Therefore, the iron-catalyzed decomposition of phospholipid hydroperoxides primarily produces epoxyperoxyl radicals, which react with the 8a-carbon centered radical of alpha-tocopherol in liposomal systems. PMID- 11934401 TI - Morphological manipulation of bolaamphiphilic polydiacetylene assemblies by controlled lipid doping. AB - Morphological transformations of bolaamphiphilic polydiacetylene (L-Glu-Bis-3) lipid assemblies from helical ribbons to vesicles and flat sheets through controlled doping are described, and the role of specific lipid dopants in these processes is discussed. Upon doping with cell surface receptor G(M1) ganglioside, fluid vesicular structures start to emerge, coexisting with the micro-crystalline helical ribbons. The vesicle formation is further facilitated and stabilized by the introduction of cholesterol into the system, presumably through surface curvature variation induced by inhomogeneous distribution and dynamic clustering of G(M1) and cholesterol within the doped assemblies. Extended helical ribbons are "truncated" into patches of flat sheets when a sufficient amount of Bis-1, a structurally compatible symmetric bolaamphiphilic diacetylene lipid, is doped. The results reaffirm the important roles of packing geometry and headgroup chirality in the formation of extended helical ribbon structures. The doped assemblies of bolaamphiphiles allow for capture of intermediate structures of morphological transformation using transmission electron microscopy (TEM). A vesicle-to-ribbon transformation mechanism via lateral reorganization within relatively fluid vesicular microstructures has been suggested. Understanding of the doping-induced transformation process provides useful information for the design of advanced materials where the microscopic morphology of material is crucial to its function. PMID- 11934402 TI - The development of ordinal numerical knowledge in infancy. AB - A critical question in cognitive science concerns how numerical knowledge develops. One essential component of an adult concept of number is ordinality: the greater than and less than relationships between numbers. Here it is shown in two experiments that 11-month-old infants successfully discriminated, whereas 9 month-old infants failed to discriminate, sequences of numerosities that descended in numerical value from sequences that increased in numerical value. These results suggest that by 11 months of age infants possess the ability to appreciate the greater than and less than relations between numerical values but that this ability develops between 9 and 11 months of age. In an additional experiment 9-month-old infants succeeded at discriminating the ordinal direction of sequences that varied in the size of a single square rather than in number, suggesting that a capacity for non-numerical ordinal judgments may develop before a capacity for ordinal numerical judgments. These data raise many questions about how infants represent number and what happens between 9 and 11 months to support ordinal numerical judgments. PMID- 11934403 TI - Object individuation using property/kind information in rhesus macaques (Macaca mulatta). AB - Around 1 year of age, infants develop the ability to individuate objects in the absence of spatiotemporal information. Some have proposed that this capacity relies on the emergence of language and, in particular, that comprehending an object's label is required to individuate it as a particular kind. One approach to testing this hypothesis is to conduct experiments on pre-linguistic human infants. A second is to test non-linguistic animals. We followed the second approach, exploring whether semi-free-ranging rhesus macaques can individuate objects using property/kind information. To make the results most directly comparable, we adapted a reaching paradigm used to examine property/kind individuation in infants. Results from three experiments demonstrate that, like 12-month-old infants, adult rhesus macaques can use both spatiotemporal and property/kind information to individuate food objects. In a fourth experiment designed to examine which properties are used to individuate food objects, results revealed that rhesus use color, but not shape. These results, together with experiments involving different procedures, provide support for the conclusion that in the absence of linguistic abilities, some non-human primates spontaneously use property/kind information to individuate objects. PMID- 11934404 TI - Turning the tables: language and spatial reasoning. AB - This paper investigates possible influences of the lexical resources of individual languages on the spatial organization and reasoning styles of their users. That there are such powerful and pervasive influences of language on thought is the thesis of the Whorf-Sapir linguistic relativity hypothesis which, after a lengthy period in intellectual limbo, has recently returned to prominence in the anthropological, linguistic, and psycholinguistic literatures. Our point of departure is an influential group of cross-linguistic studies that appear to show that spatial reasoning is strongly affected by the spatial lexicon in everyday use in a community (e.g. Brown, P., & Levinson, S. C. (1993). Linguistic and nonlinguistic coding of spatial arrays: explorations in Mayan cognition (Working Paper No. 24). Nijmegen: Cognitive Anthropology Research Group, Max Planck Institute for Psycholinguistics; Cognitive Linguistics 6 (1995) 33). Specifically, certain groups customarily use an externally referenced spatial coordinate system to refer to nearby directions and positions ("to the north") whereas English speakers usually employ a viewer-perspective system ("to the left"). Prior findings and interpretations have been to the effect that users of these two types of spatial system solve rotation problems in different ways, reasoning strategies imposed by habitual use of the language-particular lexicons themselves. The present studies reproduce these different problem-solving strategies in speakers of a single language (English) by manipulating landmark cues, suggesting that language itself may not be the key causal factor in choice of spatial perspective. Prior evidence on rotation problem solution from infants (e.g. Acredolo, L.P. (1979). Laboratory versus home: the effect of environment on the 9-month-old infant's choice of spatial reference system. Developmental Psychology, 15 (6), 666-667) and from laboratory animals (e.g. Restle, F. (1975). Discrimination of cues in mazes: a resolution of the place-vs.-response question. Psychological Review, 64, 217-228) suggests a unified interpretation of the findings: creatures approach spatial problems differently depending on the availability and suitability of local landmark cues. The results are discussed in terms of the current debate on the relation of language to thought, with particular emphasis on the question of why different cultural communities favor different perspectives in talking about space. PMID- 11934405 TI - Switching between environmental representations in memory. AB - In everyday life we accomplish tasks that require the storage and access of mental representations of different environments that we are not currently perceiving. Past research has suggested that environments are encoded by a series of independent representations that are organized in memory. Three experiments tested this idea further by asking whether multiple representations of environments can be accessed simultaneously. Using a cued task-set switching paradigm, subjects judged spatial relationships between target locations in two familiar environments. Response times were longer when successive trials probed different environments, an effect not due to switching between semantic categories or semantic priming, suggesting that representations of environments are accessed sequentially. Implications for various hypotheses concerning the properties of environmental representations are discussed. PMID- 11934406 TI - Rich interpretation vs. deflationary accounts in cognitive development: the case of means-end skills in 7-month-old infants. AB - Seven-month-old infants appear to learn means-end skills, such as pushing a button to retrieve a distant toy (Psychological Review 104 (1997) 686). The present studies tested whether such apparent means-end behaviors are genuine, or simply the repetition of trained behaviors under conditions of greatest arousal, as suggested by a dynamic systems reinterpretation. When infants were trained to repeat behaviors that did not serve as means to retrieving toys (pushing a button to light a set of distant lights), their button-pushing differed significantly from infants for whom button-pushing served as a means for retrieving toys. Further, infants demonstrated means-end skills with behaviors that they had not been trained to repeat. Implications for early means-end abilities and for debates surrounding the interpretation of infant behavior are discussed. PMID- 11934408 TI - Arithmetic operation and working memory: differential suppression in dual tasks. AB - The relationship between arithmetic function and working memory was examined using a dual-task paradigm for either phonological or visuo-spatial suppression. Simultaneous phonological rehearsal significantly delayed the performance of multiplication but not subtraction, whereas holding an image in the mind delayed subtraction but not multiplication. This result indicates that arithmetic function is related to working memory in a subsystem-specific manner: multiplication is more closely linked to phonological loop and subtraction to visuo-spatial sketchpad. Whereas this is not compatible with the notion that arithmetic is done on a unitary, amodal representation of numbers, it provides support for the triple-code and/or the modular processing models on human numerical cognition in which number representations are specific for input/output modality and arithmetic types. PMID- 11934407 TI - Enumeration of collective entities by 5-month-old infants. AB - Recent findings suggest that infants are capable of distinguishing between different numbers of objects, and of performing simple arithmetical operations. But there is debate over whether these abilities result from capacities dedicated to numerical cognition, or whether infants succeed in such experiments through more general, non-numerical capacities, such as sensitivity to perceptual features or mechanisms of object tracking. We report here a study showing that 5 month-olds can determine the number of collective entities -- moving groups of items -- when non-numerical perceptual factors such as contour length, area, density, and others are strictly controlled. This suggests both that infants can represent number per se, and that their grasp of number is not limited to the domain of objects. PMID- 11934409 TI - ESB Keynote Lecture-Dublin 2000. Outcomes of the 12th conference of the European Society of Biomechanics. PMID- 11934410 TI - Why mechanobiology? A survey article. AB - The central paradigm of skeletal mechanobiology is that mechanical forces modulate morphological and structural fitness of the skeletal tissues-bone, cartilage, ligament and tendon. Traditionally, skeletal biomechanics has focussed on how these tissues perform the structural and locomotory functions of the vertebrate skeleton. In mechanobiology the central question is how these same load-bearing tissues are produced, maintained and adapted by cells as an active response to biophysical stimuli in their environment. The idea that 'form follows function' is not new, but we now believe that the scientific community has the knowledge and tools to prove, understand and use functional adaptation to benefit medicine and human health. In this Survey Article the philosophy and progress of skeletal mechanobiology are discussed. The revival of this science, with roots dating back to the 19th Century, is now driven by new developments in cellular, molecular and computational technologies. These developments are still in an early stage of application, but if modern mechanobiology fulfills the promises of its ambitions, the results will bring great benefits to tissue engineering and to the treatment and prevention of skeletal conditions such as congenital deformities, osteoporosis, osteoarthritis and bone fractures. PMID- 11934411 TI - An introduction to biofluid mechanics--basic models and applications. AB - Cardiovascular disease is the primary cause of morbidity and mortality in the western world. Complex hemodynamics play a critical role in the development of atherosclerosis and the processes of aging, as well as many other disease processes. Biofluid mechanics play a major role in the cardiovascular system and it is important to understand the forces and movement of blood cells and whole blood as well as the interaction between blood cells and the vessel wall. Fundamental fluid mechanical, which are important for the understanding of the blood flow in the cardiovascular circulatory system of the human body aspects are presented. Measurement techniques for model studies such as LDA, ultrasound, and MRI studies will be discussed. Viscosity and flow behavior changes specifically the creation of vortices and flow disturbances can be used to show how medication can influence flow behavior. Experiments have shown that hemodynamics may have a strong influence on the creation of aneurysms and varicose veins. Other factors such as vessel wall structure are also important. In preliminary studies, it has been demonstrated that geometry and elasticity of vessel walls help determine flow behavior. High velocity fluctuations indicate flow disturbances that should be avoided. Health care practitioners must understand fluid dynamic factors such as flow rate ratio, pressure and velocity gradients, and flow behavior, velocity distribution, shear stress on the wall and on blood cells. These mechanical factors are largely responsible for the deposit of blood cells and lipids, a leading cause of atherosclerosis. The interaction between blood cells and of the cells with the vessel, leads to the formation of plaques and agglomerations. These deposits are found predominantly at arterial bends and bifurcations where blood flow is disturbed, where a secondary flow is created, and where flow separation regions are found. Experiments on hemodynamic effects in elastic silicon rubber models of the cardiovascular system with flow wire, stents, or patches for vessel surgery will be discussed. These studies can be important in improving diagnostics and therapeutic applications. PMID- 11934412 TI - Minimum measured-input models for the assessment of motor ability. AB - The problem of assessing the physical functional limitation of a given individual and establishing the relationship between impairment/s and disability using a biomechanical approach is addressed. This endeavour was pursued with reference to the locomotor system and in order to address the following specific clinical issues: prognosis, eligibility for health services, measure of the outcome of a therapy, and therapeutic programming. A thorough biomechanical analysis of selected motor tasks would be effective but awkward to apply for subject-specific evaluation in clinical practice by reason of the complexity of both instrumentation and experimental protocols. In addition, as illustrated in the paper, the adequacy of the accuracy with which this type of analysis provides relevant information may be argued. Therefore, different methods were devised in the attempt to join objectivity with field applicability. These entailed the measurement of a minimum number of biomechanical variables during the execution of the selected motor task and these quantities were acquired using a low cost experimental apparatus least perceivable to the test subject, that is a dynamometric plate. However, since data thus obtained do not necessarily lend themselves to straightforward interpretation in terms of function assessment, models of the musculo-skeletal system that embodied the invariant aspects of both the modelled system and the specific motor task were devised. Using such "minimum measured-input models", physiology-related, and thus easier to interpret, information was obtained. Two different sets of mathematical models are presented: one deals with the lowest level of detail and normally aims at assessing a global physical performance score, the other discloses joint function and segmental mechanics and therefore contributes to establishing a relationship between impairment and disability. The validation of these models, carried out in the laboratory, has shown that they possess a potential for application in clinical practice. PMID- 11934413 TI - A walking robot called human: lessons to be learned from neural control of locomotion. AB - From what we know at present with respect to the neural control of walking, it can be concluded that an optimal biologically inspired robot could have the following features. The limbs should include several joints in which position changes can be obtained by actuators across the joints. The control of mono- and biarticular actuators should occur at least at three levels: one at direct control of the actuators (equivalent to motoneuron level), the second at indirect control acting at a level which controls whole limb movement (flexion or extension) and the third at a still higher level controlling the interlimb coordination. The limb level circuits should be able to produce alternating flexion and extension movements in the limb by means of coupled oscillator flexor and extensor parts which are mutually inhibitory. The interlimb control level should be able to command the various limb control centers. All three control levels should have some basic feedback circuits but the most essential one is needed at the limb control level and concerns the decision to either flex or extend a given limb. The decision to activate the extensor part of the limb oscillator has to be based on feedback signalling the onset of loading of the limb involved. This should be signalled by means of load sensors in the limb. The decision to activate the flexor part of the limb oscillator has to depend on various types of feedback. The most important requirement is that flexion should only occur when the limb concerned is no longer loaded above a given threshold. The rule for the initiation of limb flexion can be made more robust by adding the requirement that position at the base of the limb ("hip") should be within a normal end of stance phase range. Hence, human locomotion is thought to use a number of principles which simplify control, just as in other species such as the cat. It is suggested that cat and human locomotion are good models to learn from when designing efficient walking robots. PMID- 11934414 TI - Slide track analysis of the relative motion between femoral head and acetabular cup in walking and in hip simulators. AB - Joint simulators are important tools in wear studies of prosthetic joint materials. The type of motion in a joint simulator is crucial with respect to the wear produced. It is widely accepted that only multidirectional motion yields realistic wear for polyethylene acetabular cups. Multidirectionality, however, is a wide concept. The type of multidirectional motion varies considerably between simulators, which may explain the large differences in observed wear rates. At present, little is known about the relationship between the type of multidirectional motion and wear. One illustrative way to compare the motions of various hip simulators is to compute tracks made on the counterface by selected points of the surface of the femoral head and acetabular cup due to the cyclic relative motion. A new computation method, based on Euler angles, was developed, and used to compute slide tracks for the three-axis motion of the hip joint in walking, and for two hip simulators, the HUT-3 and the biaxial rocking motion. The slide track patterns resulting from the gait waveforms were found to be similar to those produced by the HUT-3 simulator. This paper is the first to include a verification of the computed simulator tracks. The tracks were verified in the two simulators using sharp pins, embedded in acetabular cups, engraving distinct grooves onto the femoral heads. The engravings were identical to the computed tracks. The results clearly differed from earlier computations by another research group. This study is intended to start a thorough investigation of the relationship between the type of multidirectional motion and wear. PMID- 11934415 TI - Modeling the bifurcating flow in a human lung airway. AB - The inspiratory flow characteristics in a three-generation lung airway have been numerically investigated using a control volume method to solve the fully three dimensional laminar Navier-Stokes equations. The three-generation airway is extracted from the fifth to seventh branches of the model of Weibel (Morphometry of the Human Lung, Academic Press, New York, Springer, Berlin, 1963) with in plane and 90 degrees off-plane configurations. Computations are carried out in the Reynolds number range of 200-1600, corresponding to mouth-air breathing rates ranging from 0.27 to 2.16l/s, or an averaged height of a man breathing from quiet to vigorous state. Particular attention is paid to establishing relations between the Reynolds number and the overall flow characteristics, including flow patterns and pressure drop. The ratio of airflow rate through the medial branch to that of the lateral branch for an in-plane airway increases as Re(0.227). However, the total pressure drop coefficient varies as Re(-0.497) for an in-plane airway and as Re(-0.464) for an off-plane airway. These pressure drop results are in good agreement with the experimentally measured behavior of Re(-0.5) and are more accurate than the numerically determined behavior of Re(-0.61) assuming the airways to be approximated by two-dimensional channels. PMID- 11934416 TI - Mechanically induced electrical potentials of articular cartilage. AB - While there is increasing evidence that chondrocytes are affected by mechanically induced stimuli, endogenous force-related electrical potentials within articular cartilage have been so far observed only in-vitro. Using a porcine ex-vivo model (German Land Race), 8 knee joints were explanted and exposed to mechanical force (up to 800 N) using a special device. Electrodes were inserted into the cartilage matrix. With an amplifier and an A/D transducer the changes of electrical voltage between the electrodes as well as those of the force were recorded online and simultaneously on a computer. Additionally, we located one pair of electrodes on the surface of the cartilage tissue to detect electrical fields outside the cartilage tissue. In relation to the applied force we observed that electrical potentials derived from inside and outside the articular cartilage showed a correspondence. When an alternating force with an amplitude of 360 N and a frequency of about 0.2 Hz was periodically applied, we measured peak amplitudes ranging from 2.1 to 5.5 mV within the cartilage tissue with electrical negativity within the weight bearing area of the cartilage tissue. The measured voltages depended on the applied force, the location of the electrodes, and on anatomical variations. We found an almost linear relation between the magnitude of the applied force and the recorded voltage. With the help of the electrodes located outside and within the cartilage tissue, we were able to show that force dependent fields are generated inside the cartilage. There are several theories explaining the origin of these electrical phenomena, many of them focusing on the negative charges of the proteoglycans in relation to the flow of interstitial fluid and ions under compression. However, the consequences of these phenomena are yet not clear. PMID- 11934417 TI - Mechanical properties of brain tissue in tension. AB - This paper contains experimental results of in vitro, uniaxial tension of swine brain tissue in finite deformation as well as proposes a new hyper-viscoelastic constitutive model for the brain tissue. The experimental results obtained for two loading velocities, corresponding to strain rates of 0.64 and 0.64 x 10(-2)s( 1), are presented. We believe that these are the first ever experiments of this kind. The applied strain rates were similar to those applied in our previous study, focused on explaining brain tissue properties in compression. The stress strain curves are convex downward for all extension rates. The tissue response stiffened as the loading speed increased, indicating a strong stress-strain rate dependence. Swine brain tissue was found to be considerably softer in extension than in compression. Previously proposed in the literature brain tissue constitutive models, developed based on experimental data collected in compression are shown to be inadequate to explain tissue behaviour in tension. A new, non-linear, viscoelastic model based on the generalisation of the Ogden strain energy hyper-elastic constitutive equation is proposed. The new model accounts well for brain tissue deformation behaviour in both tension and compression (natural strain in <-0.3,0.2>) for strain rates ranging over five orders of magnitude. PMID- 11934418 TI - Effect of bone graft characteristics on the mechanical behavior of the lumbar spine. AB - There is little information about the influence of bone graft size, position and elasticity on the mechanical behavior of the lumbar spine. Intersegmental motion, intradiscal pressure and stresses in the lumbar spine were calculated using a three-dimensional, nonlinear finite element model which included an internal spinal fixation device and a bone graft. Cross-sectional area, position, and elastic modulus of the graft were varied in this study. Bone grafts, especially very stiff ones, increase stresses on adjacent endplates. Though larger grafts lead to less contact pressure, it is difficult to judge the quality of different bone graft positions. In general, ventral flexion results in lower maximum contact pressure than lateral bending. There is always little intersegmental rotation in the bridged region compared with that of an intact spine.A larger graft with low stiffness should be favored from a mechanical point of view. Patients should avoid lateral bending of the upper body shortly after surgery. PMID- 11934419 TI - Finite element and experimental models of cemented hip joint reconstructions can produce similar bone and cement strains in pre-clinical tests. AB - Finite element (FE) models could be used for pre-clinical testing of cemented hip replacement implants against the damage accumulation failure scenario. To accurately predict mechanical failure, the models should accurately predict stresses and strains. This should be the case for various implants. In the current study, two FE models of composite hip reconstructions with two different implants were validated relative to experimental bone and cement strains. The objective was an overall agreement within 10% between experimental and FE strains. Two stem types with different clinical results were analyzed: the Lubinus SPII and the Mueller Curved with loosening rates of 4% and 16% after 10 yr, respectively (Prognosis of total hip replacement. 63rd Annual Meeting of the American Academy of orthopaedic surgeons, Atlanta, USA). For both implant types, six stems were implanted in composite femurs. All specimens were subjected to bending. The Mueller Curved specimens were additionally subjected to torsion. Bone strains were recorded at 10 locations on the cortex and cement strains at three locations within the cement mantle. An FE model was built for both stem types and the experiments were simulated. Bone and cement strains were calculated at the experimental gauge locations. Most FE bone strains corresponded to the mean experimental strains within two standard deviations; most FE cement strains within one standard deviation. Linear regression between the FE and mean experimental strains produced slopes between 0.82 and 1.03, and R(2) values above 0.98. Particularly for the Mueller Curved, agreement improved considerably when FE strains were compared to the strains from the experimental specimen used to build the FE model. The objective of overall agreement within 10% was achieved, indicating that both FE models were successfully validated. This prerequisite for accurately predicting long-term failure has been satisfied. PMID- 11934420 TI - Cross-sectional structural parameters from densitometry. AB - Bone densitometry has previously been used to obtain cross-sectional properties of bone from a single X-ray projection across the bone width. Using three unique projections, we have extended the method to obtain the principal area moments of inertia and orientations of the principal axes at each scan cross-section along the length of the scan. Various aluminum phantoms were used to examine scanner characteristics to develop the highest accuracy possible for in vitro non invasive analysis of cross-sectional properties. Factors considered included X ray photon energy, initial scan orientation, the angle spanned by the three scans (included angle), and I(min)/I(max) ratios. Principal moments of inertia were accurate to within +/-3.1% and principal angles were within +/-1 degrees of the expected value for phantoms scanned with included angles of 60 degrees and 90 degrees at the higher X-ray photon energy (140 kVp). Low standard deviations in the error (0.68-1.84%) also indicate high precision of calculated measurements with these included angles. Accuracy and precision decreased slightly when the included angle was reduced to 30 degrees. The method was then successfully applied to a pair of excised cadaveric tibiae. The accuracy and insensitivity of the algorithms to cross-sectional shape and changing isotropy (I(min)/I(max)) values when various included angles are used make this technique viable for future in vivo studies. PMID- 11934421 TI - Measurement of a spinal motion segment stiffness matrix. AB - The six-degrees-of-freedom elastic behavior of spinal motion segments can be approximated by a stiffness matrix. A method is described to measure this stiffness matrix directly with the motion segment held under physiological conditions of axial preload and in an isotonic fluid bath by measuring the forces and moments associated with each of the six orthogonal translations and rotations. The stiffness matrix was obtained from the load-displacement measurements by linear least squares assuming a symmetric matrix. Results from a pig lumbar spinal motion segment in an isotonic bath, with and without a 500 N axial preload, showed a large stiffening effect with axial preload. PMID- 11934422 TI - An improved labelling technique for monitoring microcrack growth in compact bone. AB - Fatigue-induced damage plays an important role in bone remodelling and in the formation of stress and fragility fractures. Recently, a technique has been developed (Lee, T.C. et al., Sequential labelling of microdamage in bone using chelating agents. Journal of Orthopedic Research, 18 (2000) 322-325) which allows microcrack growth in trabecular bone to be monitored by the application of a series of chelating fluorochromes, however, some limitations were identified with the process. The aims of this study were to refine the method of detection using these agents in order to determine the optimal sequence of application and the optimal concentrations which allowed all the agents to fluoresce equally brightly using UV epifluorescence. A chemical analysis process, ion chromatography, followed by validation tests on bone samples showed that the optimal sequence of application and concentration of each agent was alizarin complexone (0.0005 M) followed by xylenol orange (0.0005 M), calcein (0.0005 M) and calcein blue (0.0001 M). A fifth agent, oxytetracycline was excluded from the study after recurring problems were found with its ability to chelate exposed calcium when applied in sequence with the other agents. This work has developed a sequential labelling technique, which allows for microcrack propagation during fatigue testing of bone specimens to be monitored without the problem of chelating agent substitution occurring. PMID- 11934423 TI - Mechanical properties of single hyaluronan molecules. AB - Hyaluronan (HA) is a major component of the extracellular matrix. It plays an important role in the mechanical functions of the extracellular matrix and stabilization of cells. Currently, its mechanical properties have been investigated only at the gross level. In this study, the mechanical properties of single HA molecules were directly measured with an optical tweezer technique, yielding a persistence length of 4.5 +/- 1.2 nm. This information may help us to understand the mechanical roles in the extracellular matrix infrastructure, cell attachment, and to design tissue engineering and drug delivery systems where the mechanical functions of HA are essential. PMID- 11934424 TI - Design and validation of a surrogate humerus for biomechanical testing. AB - At present biomechanical testing of fracture plating strategies is conducted using animal or cadaveric whole bone models. This may introduce experimental error into these studies. This communication summarises the design and validation of a novel bone and fibre-reinforced plastic construct conceived to minimise intra-experimental error. A tubular surrogate humerus was produced with dimension and strength matched to that of the human humerus. Bone inserts placed into the wall of the tube allow for the fixation of the plates with bone screws. Three point bending tests of the flexural rigidity of the surrogate humerus (EI=100.1 (SD 6.0)Nm(2)) showed it to be comparable to the human humerus. Further, pull-out tests of the screws showed that the bone slots adequately mimicked the whole bone scenario. This testing construct will be used for a comparative study of humeral plating techniques. PMID- 11934425 TI - Accelerometer and rate gyroscope measurement of kinematics: an inexpensive alternative to optical motion analysis systems. AB - A general-purpose system to obtain the kinematics of gait in the sagittal plane based on body-mounted sensors was developed. It consisted of four uniaxial seismic accelerometers and one rate gyroscope per body segment. Tests were done with 10 young healthy volunteers, walking at five different speeds on a treadmill. In order to study the system's accuracy, measurements were made with an optic, passive-marker system and the body-mounted system, simultaneously. In all the comparison cases, the curves obtained from the two systems were very close, showing root mean square errors representing <7% full range in 75% of the cases (overall mean 6.64%, standard deviation 4.13%) and high coefficients of multiple correlation in 100% of cases (overall mean 0.9812, standard deviation 0.02). Calibration of the body-mounted system is done against gravity. The body mounted sensors do not hinder natural movement. The calculation algorithms are computationally demanding and only are applicable off-line. The body-mounted sensors are accurate, inexpensive and portable and allow long-term recordings in clinical, sport and ergonomics settings. PMID- 11934426 TI - ISB recommendation on definitions of joint coordinate system of various joints for the reporting of human joint motion--part I: ankle, hip, and spine. International Society of Biomechanics. AB - The Standardization and Terminology Committee (STC) of the International Society of Biomechanics (ISB) proposes a general reporting standard for joint kinematics based on the Joint Coordinate System (JCS), first proposed by Grood and Suntay for the knee joint in 1983 (J. Biomech. Eng. 105 (1983) 136). There is currently a lack of standard for reporting joint motion in the field of biomechanics for human movement, and the JCS as proposed by Grood and Suntay has the advantage of reporting joint motions in clinically relevant terms. In this communication, the STC proposes definitions of JCS for the ankle, hip, and spine. Definitions for other joints (such as shoulder, elbow, hand and wrist, temporomandibular joint (TMJ), and whole body) will be reported in later parts of the series. The STC is publishing these recommendations so as to encourage their use, to stimulate feedback and discussion, and to facilitate further revisions. For each joint, a standard for the local axis system in each articulating bone is generated. These axes then standardize the JCS. Adopting these standards will lead to better communication among researchers and clinicians. PMID- 11934427 TI - Mechanical properties of cancellous bone in the human mandibular condyle are anisotropic. PMID- 11934429 TI - Problems in application of purely linear models in cerebral circulation. PMID- 11934432 TI - Cellular communication and bystander effects: a critical review for modelling low dose radiation action. AB - Available data suggesting the occurrence of "bystander effects" (i.e. damage induction in cells not traversed by radiation) were collected and critically evaluated, in view of the development of low-dose risk models. Although the underlying mechanisms are largely unknown, cellular communication seems to play a key role. In this context, the main features of cellular communication were summarised and a few representative studies on bystander effects were reported and discussed. Three main approaches were identified: (1) conventional irradiation of cell cultures with very low doses of light ions; (2) irradiation of single cells with microbeam probes; (3) treatment with irradiated conditioned medium (ICM), i.e. feeding of unexposed cells with medium taken from irradiated cultures. Indication of different types of bystander damage (e.g. cell killing, gene mutations and modifications in gene expression) has been found in each of the three cases. The interpretations proposed by the investigators were discussed and possible biases introduced by specific experimental conditions were outlined. New arguments and experiments were suggested, with the main purpose of obtaining quantitative information to be included in models of low-dose radiation action. Implications in interpreting low-dose data and modelling low-dose effects at cellular and supra-cellular level, including cancer induction, were analysed. Possible synergism with other low-dose specific phenomena such as adaptive response (AR) (i.e. low-dose induced resistance to subsequent irradiation) was discussed. PMID- 11934433 TI - Induction of stem cell cycling in mice increases their sensitivity to a chemical leukaemogen: implications for inherited genomic instability and the bystander effect. AB - Preconception paternal irradiation (PPI) modifies haemopoietic and stromal tissues of offspring and increases risk of generating lympho-haemopopietic malignancy if those offspring are then exposed to a leukaemogen. We hypothesised that this increased risk was related to inherited damage which had caused increased stem cell proliferation rates. To test for this link, in vivo, rapid stem cell proliferation was established by giving sub-lethal irradiation (3Gy gamma-rays) and allowing 3 days recovery. At this stage, 60% of haemopoietic spleen colony-forming units (CFU-S) were in DNA-synthesis, compared to <10% in unirradiated controls. Two groups of mice, unirradiated controls and irradiated animals, were then injected with 50mg/kg methyl nitrosourea (MNU) and observed daily for onset of lympho-haemopoietic malignancy. In a further control group of 60 mice, irradiated but not injected with MNU, only one leukaemia developed. In unirradiated controls, 20% of the mice developed malignancies between 3 and 8 months later: in the irradiated, MNU-treated groups, 95% developed malignancies between 2 and 7 months later. Thus, at least one powerful potentiating mechanism for induction of lympho-haemopoietc malignancy following inherited damage can be related to haemopoietic stem cell proliferation. Genomic instability is exposed by cell proliferation and has been implicated in this type of damage. However, a regulatory stromal microenvironment plays a part in inducing that proliferation. Thus, the microenvironment is the effective "bystander" which is thought to promote and amplify genomic instability, and thereby influence the induction of malignancy both in PPI offspring and in mice with induced stem cell proliferation. PMID- 11934434 TI - A high frequency of mtDNA polymorphisms in HeLa cell sublines. AB - The complete mtDNA sequences from the uncloned "founder" HeLa cells and from five sublines have been determined. These sequences all carry a common "core" of 38 single basepair alterations relative to the revised Cambridge Reference Sequence (CRS). The HeLa mitochondrial genome is of African descent and it is a member of the African L3 haplogroup. The sequence of the HeLa mtDNA resolves the uncertainty surrounding the mosaic composition of the original CRS for human mtDNA. Most importantly, we detected a total of eight polymorphisms that have arisen in the mtDNA coding region of different HeLa sublines. These observations suggest that HeLa mtDNA has a high rate of sequence divergence, relative to the phylogenetically-derived divergence rate for mtDNAs in the human population, which results from a relaxation of negative selection against the fixation of deleterious mutations. Furthermore, this high frequency of polymorphisms in HeLa mtDNA may reflect a process similar to the accumulation of somatic mtDNA mutations in human cancers. Preliminary analysis of single-cell derived subclone lines revealed the occurrence of another polymorphism and provided evidence for a large number of mtDNA segregation units. PMID- 11934435 TI - Dietary modulation of mitochondrial DNA deletions and copy number after chemotherapy in rats. AB - Mitochondrial DNA (mtDNA) is particularly susceptible to mutation by alkylating agents, and mitochondrial damage may contribute to the efficacy and toxicity of these agents. We found that folate supplementation decreased the frequency of the "common deletion" (4.8kb, bases 8103-12,936) in liver from untreated rats and from animals treated with cyclophosphamide but not 5-fluorouracil (5-FU). The relative abundance of mitochondrial DNA was greater after chemotherapy but there was no effect of diet. Rats fed with a purified diet had fewer mitochondrial deletions than those maintained on a cereal-based diet after chemotherapy. These results indicate that diet can modulate the extent of mitochondrial damage after cancer chemotherapy, and that folic acid supplementation may be protective against mitochondrial DNA deletions. PMID- 11934436 TI - Spontaneous frequency of exon skipping in the human HPRT gene. AB - In order to elucidate the mechanisms of mRNA splicing fidelity and the mutagenic potential of aberrant mis-spliced transcripts we have investigated the frequency of spontaneous exon skipping in the human hypoxanthine-guanine phosphoribosyl transferase (HPRT) gene in well characterized human primary fibroblasts isolated from two different individuals. In these cells, coexisting with the WT species, we also detected three aberrant HPRT transcripts missing exon IV, VII, or VIII. We were unable to detect transcripts missing exon II, III, V, or VI. Significantly, all the exons affected by skipping do not generate new stop codons more than 50 nucleotides upstream from the 3' most exon-exon junction. Exon VIII was the most prone to skipping with a relative frequency to WT of 0.019+/-0.004 (approximately one aberrant transcript per 50 WT transcripts). Exon IV exhibited a relative frequency of skipping of 0.006+/-0.002 ((approximately one aberrant transcript per 150 WT transcripts) and exon VII exhibited a relative frequency of skipping of 0.003+/-0.002 ((approximately one aberrant transcript per 300 WT transcripts). These data demonstrate that aberrant transcripts with exon skipped are generated spontaneously in humans and some appear to persist in the cell. PMID- 11934437 TI - Glycophorin A mutant frequency in radiation workers at the nuclear power plants and a hospital. AB - We studied to assess the validity of the glycophorin A (GPA) mutant assay as a biological marker of the cumulative effects of chronic low doses of ionizing radiation. In 144 nuclear power plants workers and 32 hospital workers, information on confounding factors, such as age and cigarette smoking, was obtained through a self-administered questionnaire. The information on physical exposure doses was obtained from the registries for radiation exposure monitoring and control at each facility. The range of cumulative exposure doses were 0 12.02cGy. GPA mutant assay was performed by the BR6 method with modification using a FACScan flow cytometer. Potential confounders, such as, age and cigarette smoking habits showed increasing trends with GPA variants, but were not of statistical significance. The hospital workers showed higher frequency of the GPA NO variant than nuclear power plant workers. Significant dose-response relationships were found between cumulative exposure to radiation and variants levels by simple and multiple linear regression models. The slope of regression equation of the dose-response of nuclear power plants workers was much smaller than that of hospital workers. These findings suggest that there may be dose-rate effects. In a population exposed to chronic low-dose radiation, the GPA assay shows potential to be used as an effective biologic marker for assessing the cumulative exposure dose although it could not be able to see a dose relation below 10cGy of cumulative exposure dose. PMID- 11934438 TI - Role of P53 functionality in the genotoxicity of metronidazole and its hydroxy metabolite. AB - P53 mediates several biological processes for preservation of genetic stability such as the induction of cell cycle arrest, DNA repair or apoptosis in response to DNA damage. The antiparasitic drug, 1-(2-hydroxyethyl)-2-methyl-5 nitroimidazole (metronidazole, MTZ) is able to increase lymphocyte proliferation inducing at the same time chromosomal aberrations. Trying to understand this unexpected event we used cell lines with different P53 functionality, determining the proliferation capacity and the induction of micronuclei (MN) after the treatment with MTZ or its hydroxy metabolite. Our results show that MTZ increased proliferation in a dose response manner in all P53 functional cell lines without inducing changes on the levels of P53 nor MN. However, MTZ hydroxy metabolite induced a dose response increase of P53 and MN, while cell proliferation was not increased. Several studies have shown that the hydroxy metabolite is more potent than MTZ itself. Only in cell lines that do not have a functional P53, MTZ and its metabolite increased both cell proliferation and MN. MTZ use is increasing and its carcinogenicity has not been discarded. Our data indicate that MTZ hydroxy metabolite is potentially a carcinogen and needs to be further studied. PMID- 11934439 TI - Association of DNA-protein crosslinks and breast cancer. AB - This study examined the possible effect of cytochrome P450 (CYP1A1), glutathione S-transferase (GSTM1 and T1) and N-acetyltransferases 2 (NAT2) polymorphisms on DNA-protein crosslinks (DPC) formation in the white blood cells of breast cancer patients, and assessed the levels of DPC detected. Sixty cases of breast cancer were examined, all involving women diagnosed with primary, histopathologically confirmed breast cancer at the Chinese Medical College Hospital in central Taiwan. Additionally, 60 healthy women without breast cancer were selected as a control group, matched by age, cigarette smoking habits, and history of breast cancer among first-degree relatives. Known risk factors for breast cancer, including menarche before 13 years of age (OR=3.2; CI, 1.1-9.5), no history of breast-feeding (OR=4.7; CI, 1.5-14.4) and use of oral contraceptives (OR=9.1; CI, 2.8-29.8), were found to be significantly associated with breast cancer. For the CYP1A1 MspI polymorphism, 16.7 and 18.3% of cases and controls, respectively contained both alleles with the MspI restriction fragment length polymorphism (RFLP). Regarding the NAT2 allele, 25.0 and 21.7% of cases and controls carried slow genotypes. For GSTM1 and GSTT1, 56.7 and 45.0% of cases, as well as 58.3 and 43.3% of controls, contained the null genotype. Meanwhile, chi(2)-tests found no significant differences between the groups. After controlling for confounders such as cigarette smoking and family history of breast cancer, the DPC value of the case group significantly exceeded that of the control group (1.62% versus 0.98%, P<0.001). In conclusion, our findings were inconsistent with those of previous studies that showed polymorphism genes (CYP1A1, NAT2, GSTM1 and GSTT1) were associated with cancer risk. However, this study indicated that genotypic variants of these polymorphisms did not elevate the risk for breast cancer, individually or interactively. Additionally, this investigation represents the first description of the use of DPC as a biomarker to assess the level of DNA damage of breast cancer patients. Our data suggest that the DPC method is a useful tool for detecting DNA damage, and DPC formation may be associated with the induction of breast cancer. PMID- 11934440 TI - Salmonella typhimurium mutagenicity tester strains that overexpress oxygen insensitive nitroreductases nfsA and nfsB. AB - We have designed and constructed a series of plasmids that contain the major and/or minor Escherichia coli nitroreductase genes, nfsA and nfsB, in different combinations with R plasmid mucA/B genes and the Salmonella typhimurium OAT gene. The plasmid encoded gene products are necessary for both the metabolic activation of a range of structurally diverse nitrosubstituted compounds, and for mutagenic translation bypass. Introduction of these plasmids into S. typhimurium TA1538 and TA1535 has created several new tester strains which exhibit an extremely high mutagenic sensitivity and a broad substrate specificity towards a battery of nitrosubstituted test compounds that included 4-nitroquinoline-1-oxide (4-NQO), nitrofurazone (NF), 1-nitropyrene (1-NP), 2-nitronaphthalene (2-NN), 2 nitrofluorene (2-NF), and 1,6-dinitropyrene (1,6-DNP). Our studies show that the nfsA gene encodes a product that is extremely effective in the metabolic activation of a range of structurally diverse nitrosubstituted compounds. Several of the new tester strains are more than two orders of magnitude more sensitive to nitrosubstituted compounds than the Ames tester strains TA100 or TA98. In addition to enhancing mutagenic sensitivity, plasmids encoding both metabolic and mutagenesis functions on a single plasmid provide considerable flexibility for future mechanistic studies or tester strain development, in which it may be necessary to introduce additional plasmids containing different antibiotic resistance markers. PMID- 11934441 TI - Analysis of variation in the human beta-globin gene cluster using a novel DHPLC technique. AB - We have implemented a technique combining allele-specific PCR (AS-PCR) and denaturing high-performance liquid chromatography (DHPLC) to identify new polymorphic variants within an intergenic region in the beta-globin cluster. This technique is applicable to the detection of new variants in genomic regions where variation is apportioned into distinct classes of haplotype. Duplexes for DHPLC analysis were created by denaturation and re-annealing of a mixture of two AS-PCR products of known and unknown sequence from the same haplotypic class, permitting detection of new haplotypes in each class. A 454bp fragment 3.5kb 5' to the human delta-globin gene, which may have a gene regulatory function, was analysed in 840 chromosomes from a global sampling of human populations using this method. Two divergent haplotypes were found to predominate in all populations studied, possibly as a result of balancing selection. PMID- 11934442 TI - The effect of DNA damage on the formation of protein/DNA complexes. AB - Many cellular functions including gene expression and chromosome structure are highly dependent upon the precise recognition and binding of specific DNA elements by regulatory and structural proteins. DNA damage that alters protein/DNA interactions therefore has the potential to disrupt normal cellular functions including proliferation. As a model to examine the interaction of proteins with damaged DNA, the binding of AP-1 transcription factor to cognate DNA elements with 8-oxoadenine, 8-oxoguanine and abasic sites was studied by gel mobility shift analysis. Of the three types of DNA damage only 8-oxoadenine was without effect on AP-1 binding. A single 8-oxoguanine could partially inhibit AP 1 binding when located at specific positions within and even adjacent to the conserved AP-1 binding sequence. Abasic site damage also demonstrated a position effect but with more overall inhibition. When 8-oxoguanine and abasic sites were combined to model the multiple damage sites produced by ionizing radiation there was a cumulative loss of AP-1 binding that appeared to be synergistic. These results suggest protein/DNA interactions can be quite sensitive to the site, degree, and type of DNA damage, even relatively minor modifications. PMID- 11934443 TI - DNA-adducts and atherosclerosis: a study of accidental and sudden death males in the Czech Republic. AB - Atherosclerosis and carcinogenesis may share some common mechanisms of the genotoxic action of exogenous compounds, such as polycyclic aromatic hydrocarbons (PAHs). The main objective of this study was to test the hypothesis that "bulky" aromatic DNA-adducts in smooth muscle cells (SMCs) of thoracic aortas taken at autopsy from sudden and accidental death male subjects, aged between 30 and 60 years (N=133), are associated with the stage of atherosclerosis. The subjects with severe atherosclerotic damage were treated as "Cases" (N=66). The subjects meeting diagnostic criteria for slight and moderate total atherosclerotic body damage were treated as "Controls" (N=67). An additional objective of the study was to evaluate the effect of known atherogenic risk factors and possible modifiers of atherosclerotic changes, such as age, smoking, plasma lipid and antioxidant vitamin levels and some genetic susceptibility markers, e.g. polymorphisms of GSTM1, GSTT1, NAT2, CYP1A1 or apolipoprotein E (APO E) genes. We found significantly higher DNA-adduct levels in "Cases" as compared with "Controls" (2.11+/-1.07 adducts/10(8) nucleotides versus 1.49+/-0.55 adducts/10(8) nucleotides, P<0.001). "Cases" were significantly older and had elevated heart weight and plasma cholesterol levels and a higher frequency of overweight subjects as compared with "Controls". No significant differences in DNA-adduct levels between smokers and non-smokers within either group were detected. Multivariate logistic regression revealed that the "bulky" aromatic DNA adducts, which are the most likely related to environmental exposure to genotoxic chemicals, remain a statistically significant predictor of the stage of atherosclerosis (OR=3.76, 95% CI=1.54-9.18, P=0.004) even after adjustment for age, smoking, obesity, heart weight and genetic susceptibility markers (GSTT1 and CYP1A1-MspI polymorphisms) that were also significant predictors. The fact that the "bulky" aromatic DNA-adduct levels predict the progression of atherosclerosis independently of smoking indicates that the formation of atherosclerotic plaques may also be initiated by environmental exposures other than tobacco smoke. PMID- 11934444 TI - Asymmetry of frameshift mutagenesis during leading and lagging-strand replication in Escherichia coli. AB - Mutations in DNA, including frameshifts, may arise during DNA replication as a result of mistakes made by the DNA polymerase in copying the DNA template strands. In our efforts to better understand the factors that contribute to the accuracy of DNA replication, we have investigated whether frameshift mutations on the Escherichia coli chromosome occur differentially within the leading and lagging-strands of replication. The experimental system involves measurement of the reversion frequency for several defined lac frameshift alleles in pairs of strains in which the lac target is oriented in the two possible directions relative to the origin of chromosomal replication. Within these pairs any defined lac sequence will be subject to leading-strand replication in one orientation and to lagging-strand replication in the other. Fidelity differences between the two modes of replication can be observed as a differential lac reversion between the two strains. Our results, obtained with a series of lac alleles in a mismatch repair-defective background, indicate that for at least some of the alleles there is indeed a difference in the fidelity of replication between the two modes of replication. PMID- 11934445 TI - PROGINS Alu insertion and human genomic diversity. AB - A polymorphic Alu element belonging to the young Ya5 subfamily of Alu repeats located in the progesterone receptor gene has been characterized. Using a polymerase chain reaction (PCR)-based assay, the genetic diversity associated with the PROGINS Alu repeat was determined in a diverse array of human populations. The level of insertion polymorphism associated with PROGINS suggests that it will be a useful marker for the study of human evolution. In addition, we determined the distribution of the PROGINS Alu insertion in two groups of women from greater New Orleans, LA with breast cancer. The PROGINS Alu insertion was not associated with breast cancer in the populations tested. PMID- 11934446 TI - The effect of retinoids and butyrate on the expression of CRX and IRBP in retinoblastoma cells. AB - We sought to determine whether differentiation agents such as retinoids and butyrate regulate transcript levels of interphotoreceptor retinoid binding protein (IRBP) and cone rod homeobox (CRX), a homeodomain transcription factor that regulates IRBP promoter activity. WERI-Rb1 retinoblastoma cells were treated with all-trans retinol, all-trans retinoic acid, or butyrate. IRBP and CRX mRNA levels were determined by quantitative RT-PCR. Butyrate at low concentrations increased both mRNA levels but suppressed them at higher concentrations. Retinoic acid had minimal effects. Retinol increased CRX mRNA over four fold. IRBP and CRX transcript levels are sensitive to butyrate and CRX expression is sensitive to retinol. PMID- 11934447 TI - Sensitivity to global form in glass patterns after early visual deprivation in humans. AB - To compare the effects of early monocular versus early binocular deprivation on the perception of global form, we assessed sensitivity to global concentric structure in Glass patterns with varying ratios of paired signal dots to noise dots. Children who had been deprived by dense congenital cataracts in one (n=10) or both (n=8) eyes performed significantly worse than comparably aged children without eye problems. Consistent with previous results on sensitivity to global motion [Vision Research 42 (2002) 169], thresholds in the deprived eyes were significantly better after monocular deprivation than after binocular deprivation of comparable duration, even when there had been little patching of the nondeprived eye after monocular deprivation. Together, the results indicate that the competitive interactions between a deprived and nondeprived eye evident in the primary visual cortex can co-occur with complementary interactions in extrastriate cortex that enable a relative sparing of some visual functions after early monocular deprivation. PMID- 11934448 TI - Covert attention affects the psychometric function of contrast sensitivity. AB - We examined the effect of transient covert attention on the psychometric function for contrast sensitivity in an orientation discrimination task when the target was presented alone in the absence of distracters and visual masks. Transient covert attention decreased both the threshold (consistent with a contrast gain mechanism) and, less consistently, the slope of the psychometric function. We assessed performance at 8 equidistant locations (4.5 degrees eccentricity) and found that threshold and slope depended on target location-both were higher on the vertical than the horizontal meridian, particularly directly above fixation. All effects were robust across a range of spatial frequencies, and the visual field asymmetries increased with spatial frequency. Notwithstanding the dependence of the psychometric function on target location, attention improved performance to a similar extent across the visual field.Given that, in this study, we excluded all sources of external noise, and that we showed experimentally that spatial uncertainty cannot explain the present results, we conclude that the observed attentional benefit is consistent with signal enhancement. PMID- 11934449 TI - Fast long-distance interactions in the early processing of motion-defined form and of combinations of motion-defined, luminance-defined, and cyclopean form. AB - Humans can compare the orientations and locations of two motion-defined test bars several degrees apart so as to rapidly encode and place in memory their mean orientation, orientation difference, separation and mean location, while ignoring stimuli located between the two test bars. Performance is not impaired by randomly varying the location of the bars. We conclude that the two test bars are not compared by shifting gaze location or attention from one test bar to the other, nor by attending to two spatial locations. In addition, observers can discriminate the orientation difference and mean orientation of two test bars that, each of which is rendered visible by a different sub-modality (motion, disparity or luminance). Taking into account the findings reported here and previously reported findings on the early processing of luminance-defined form (Vis. Res. 40 (2000) 2291, Vis. Res. 42 (2002) 49) and cyclopean form (Proc. Roy. Soc. Lond. B 268 (2001) 213) we propose that the human visual system contains a fast long-distance comparator that compares the orientation and locations of two test bars while being insensitive to stimuli in the space between the test bars, and that this process is independent of whether the test bars are rendered visible by only one of three kinds of contrast (luminance, disparity, motion) or by combinations of the three. One role of this comparator mechanism may be to rapidly bind the spatial aspects of the retinal image across sub-modalities immediately after each saccade. PMID- 11934451 TI - Self-organized pattern formation: experimental dissection of motion detection and motion integration by variation of attentional spread. AB - The formation of global motion patterns depends on the stimulus activation of local motion detectors as well as integrative excitatory and/or inhibitory interactions among the activated detectors. The counterphase row-of-elements [Vis. Res. 34 (1994) 1843] is an ideal stimulus for examining the relationship between the activational/energizing effect of the stimulus and interaction among the activated detectors. This is because the formation of the alternative unidirectional and oscillatory motion patterns for this stimulus requires the stimulation of local motion detectors, but there is no information in the stimulus that specifies either of the patterns. Their formation depends instead on the relative contributions of excitatory and inhibitory interactions to detector activation; the temporal patterns are self-organized. Broadly spread attention affects motion integration by changing the balance of excitatory versus inhibitory interactions, increasing the perception of unidirectional compared with oscillatory motion. (It likewise increases the perception of group compared with element motion for the Ternus stimulus.) There is, however, little if any effect of attentional spread on the luminance contrast required for the perception of single-element motion. The results indicate that the balance of integrative excitatory and/or inhibitory detector interactions can be modified by the perceiver's spread of attention, and further, that such changes need not be mediated by changes in the local, stimulus activation of the detectors. PMID- 11934450 TI - What limits detection and resolution of short-wavelength sinusoidal gratings across the retina? AB - Peripheral resolution acuity for achromatic gratings is known to be limited by the density of the underlying ganglion cell sampling array. After confirming isolation of the short-wavelength sensitive (SWS) system using chromatic adaptation methods, we wished to determine if resolution is limited by blue/yellow ganglion cell sampling (evidenced by a superiority of detection over resolution acuity and the perception of aliasing) and thus directly related to SWS-driven ganglion cell density. We measured detection and resolution acuity between 0 degrees and 35 degrees, using blue sinusoidal gratings superimposed on a yellow adapting background which ranged in intensity from 2.5 to 4.7logTrolands. At all locations, a break could be observed in the acuity vs. illumination curves followed by a plateau. Detection and resolution acuity were the same for low background intensities, but resolution acuity was lower than detection at higher intensities, accompanied by observations of chromatic aliasing. SWS resolution is sampling-limited across the retina and agrees well with predicted performance based on anatomical estimates of small bistratified ganglion cell density. PMID- 11934452 TI - Perceptual grouping in the Ternus display: evidence for an 'association field' in apparent motion. AB - We present psychophysical experiments designed to reveal the role of facilitative contour interactions (the so-called 'association field') in apparent motion. We use the Ternus display (a trio of horizontally aligned elements oscillating in apparent motion). This display is perceived in 'element' motion when interframe intervals (IFIs) are short, and in 'group' motion when IFIs are long. Using Gabor elements arranged collinearly or in parallel, IFI is varied to find group motion thresholds. Consistent with a role for collinearity in perceptual grouping, thresholds are lower for collinear displays. The collinear vs. parallel comparison is made while manipulating contrast, spatial frequency, eccentricity, phase, orientation jitter and element separation. Results show a clear effect of contrast not observed in lateral masking paradigms or in 'pathfinder' stimuli, with higher contrast promoting within-frame grouping, and evidence of facilitatory interactions among parallel elements (although over a smaller scale). The tendency for collinear displays to group more than parallel displays declined with eccentricity with no clear difference evident at 12 deg. These changes in group motion thresholds indicate changing association strengths among the elements and is accounted for in terms of an association field. Alternative accounts in terms of second-order collector units or visible persistence are considered but are not supported by the data. PMID- 11934453 TI - On the classification of simple and complex cells. AB - In their pioneering studies of primary visual cortex, Hubel and Wiesel described the existence of two classes of cells, which they termed "simple" and "complex". The original classification scheme was based on a number of partly subjective tests of linear spatial summation. Later, investigators adopted an objective classification method based on the ratio between the amplitude of the first harmonic of the response and the mean spike rate (or the F(1)/F(0) ratio) when the neuron is stimulated with drifting sinusoidal gratings. This measure is bimodally distributed over the population and divides neurons into two classes that correspond closely to the classical definition by Hubel and Wiesel. Here we show that a simple rectification model can predict the observed bimodal distribution of F(1)/F(0) in primary visual cortex when the distributions of the intracellular response modulation and mean are unimodal. Thus, contrary to common belief, the bimodality of F(1)/F(0) does not necessarily imply the existence of two discrete cell classes. Furthermore, in reviewing the literature, we find no independent support for a simple/complex dichotomy. These results suggest that the existence of two distinct neural populations in primary visual cortex, and the associated hierarchical model of receptive field organization, need to be re evaluated. PMID- 11934454 TI - A model of speed tuning in MT neurons. AB - We have shown previously that neurons in the middle temporal (MT) area of primate cortex have inseparable spatiotemporal receptive fields-their response profiles exhibit a ridge that is oriented in the spatiotemporal frequency domain, and this orientation predicts the neurons' preferred speed. When measured in spatiotemporal frequency space, such MT spectral receptive field (SRF) properties are closely matched to the spectrum generated by a moving edge. In contrast, V1 neurons have SRF properties that are poorly matched to moving edge spectra, indicating that V1 neurons are not tuned to a particular image speed but rather to specific spatial and temporal frequencies. Here we describe a neural mechanism based directly on the properties of V1 neurons that is able to explain the SRF change that occurs between V1 and MT. We outline the theory behind this transformation and posit an explanation for how the visual system extracts true speed (independent of spatial frequency) from retinal image motion. We tested this speed model against our MT neuron data and found that it provides an excellent account of speed tuning in MT. PMID- 11934455 TI - Mapping of retinal function in Best macular dystrophy using multifocal electroretinography. AB - In order to evaluate the function of the retina in Best macular dystrophy (BMD) 18 patients were examined by means of the multifocal electroretinogram (mfERG). The mfERG peak amplitudes of the central and pericentral responses were significantly reduced in the BMD patients (p<0.001). The ERG amplitude decrease of the central response was significantly correlated with visual acuity loss and with the funduscopic staging. The implicit times in more eccentric groups were slightly but significantly increased. The markedly reduced mfERG amplitudes with only slightly increased implicit times may indicate cone photoreceptor cell loss or damage to the cone outer segments. PMID- 11934456 TI - Wavefront aberrations in eyes of emmetropic and moderately myopic school children and young adults. AB - Wavefront aberrations were measured using a psychophysical ray-tracing technique in both eyes of 316 emmetropic and moderately myopic school children and young adults. Myopic subjects were found to have greater mean root mean square (RMS) value of wavefront aberrations than emmetropic subjects. Emmetropic adults had the smallest mean RMS, which remained smaller than the values for myopic adults and children and for emmetropic children both when second order Zernike aberrations (astigmatism) and third order Zernike aberrations were removed. Twenty percent of myopic adults had RMS values greater than values for all of the emmetropic adults, with significantly greater values for Zernike aberrations from second to seventh orders. High amounts of wavefront aberrations, which degrade the retinal image, may play a role in the development of myopia. PMID- 11934458 TI - The application of PCR-ELISA to the detection of Trypanosoma brucei and T. vivax infections in livestock. AB - Teneral tsetse flies infected with either Trypanosoma brucei or T. vivax were fed on healthy cattle. Blood samples collected daily from the cattle were examined by microscopy for the presence of trypanosomes, in thick smear, thin smear and in the buffy coat (BC). All the cattle fed upon by infected tsetse developed a fluctuating parasitaemia. DNA was extracted from the blood of these cattle and subjected to polymerase chain reaction (PCR) using oligonucleotide primers specific for T. brucei or T. vivax. The PCR products unique to either T. brucei or T. vivax were identified following amplification of DNA from the blood samples of infected cattle, whereas none was detectable in the DNA from the blood of the cattle exposed to non-infected teneral tsetse. In a concurrent set of experiments, one of the oligonucleotide primers in each pair was biotinylated for use in PCR-ELISA to examine all the blood samples with this assay. Both the PCR and the PCR-ELISA revealed trypanosome DNA in 85% of blood samples serially collected from the cattle experimentally infected with T. brucei. In contrast, the parasitological assays showed trypanosomes in only 21% of the samples. In the blood samples from cattle experimentally infected with T. vivax, PCR and PCR ELISA revealed trypanosome DNA in 93 and 94%, respectively. Microscopy revealed parasites in only 63% of the BCs prepared from these cattle. Neither PCR nor PCR ELISA detected any trypanosome DNA in blood samples collected from the animals in the trypanosome-free areas. However, both assays revealed the presence of trypanosome DNA in a number of blood samples from cattle in trypanosomosis endemic areas. PMID- 11934459 TI - Increase in milk yield following eprinomectin treatment at calving in pastured dairy cattle. AB - Gastrointestinal nematodes rarely cause signs of clinical disease in adult cattle. However, they have been shown to exert a negative impact on production in lactating animals, as seen by improved production following elimination of the worms using anthelmintics. A double blind, randomized clinical trial was performed in 28 dairy herds in Canada. The objective of the study was to evaluate the effect of treatment with eprinomectin pour-on solution (IVOMEC EPRINEX) at calving on production, in cattle that have had some exposure to pasture. Cows were randomly allocated to treatment or placebo in blocks of 10, based on calving date, and treated with eprinomectin or placebo on the day of calving. Information on milk production was obtained from all animals, as well as recorded cases of selected diseases. Milk production results from the Canadian dairy herd management system database were analysed using a mixed model with herd as a random effect and test within-cow as a repeated measurement. Test day milk yields from the first six tests after treatment were included in the model, representing a period of between 180 and 200 days in milk (dim). Treated cows produced an additional 0.94 kg of milk per day when compared to the controls over this period. The production effect was independent of calving season, age of the animal and geographical location. No effect of treatment was seen on milk composition, somatic cell count (scc) or on the selected health parameters that were recorded for all included animals. Monthly fecal egg counts (FEC) were performed for eight randomly selected animals in each herd. The observed FEC were low in this study, with a range from 0 to 419 trichostrongyle type eggs per 5g (ep5g) of feces in animals not yet treated with the anthelmintic. The average count was 9.8 and the median was 1.0. FECs dropped immediately after calving and stayed lower for at least 100 days in treated animals when compared to controls. In conclusion, gastrointestinal nematodes appear to have an effect on milk production in Canadian dairy cows that have had some degree of pasture exposure. Eliminating the present subclinical parasite burdens produced a consistent increase in milk production that can yield economic benefits for the dairy producer. PMID- 11934461 TI - Infectivity of Trichinella papuae for experimentally infected red foxes (Vulpes vulpes). AB - To evaluate infectivity for carnivores as well as other biological characteristics of the newly described Trichinella papuae, eight red foxes were experimentally infected with the parasite. Five weeks after inoculation, T. papuae larvae were recovered from nine different muscle types. The larvae recovered from muscle tissue were shown to be infective to mice, to have a very low tolerance to freezing, and to survive longer than the other Trichinella genotypes in decaying tissue up to 5 weeks after infection. PMID- 11934460 TI - Larval toxocarosis in sheep: the immunohistochemical characterization of lesions in some affected organs. AB - Morphological and immunohistochemical responses of lambs following oral infection with 10,000 infective Toxocara canis (T. canis) eggs were studied up to 28 days post-infection (pi). The small intestine, liver, lungs and brain of both infected and control lambs were examined using the routine histological methods for paraffin sections and immunohistochemical techniques for frozen tissue sections. Eosinophil-rich hepatic granulomas and diffuse T. canis-induced pulmonary inflammation were the most prominent pathological features. CD2+, CD4+, CD8+, IgM bearing cells, and macrophages were demonstrated in the liver granulomas. The observed histomorphologic changes were similar to other paratenic hosts. It was concluded that larval toxocarosis followed the classical migratory path in the infected lambs. PMID- 11934462 TI - Use of lectin binding characteristics to identify gastrointestinal parasite eggs in faeces. AB - Identification of the presence and abundance of species is important when choosing therapies and control strategies for internal parasitism of livestock. Here we examine lectin binding characteristics of eggs isolated from sheep faeces as a means for identifying the parasite genera contributing to infection. The intensity of lectin staining varied with incubation time, incubation volume, concentration of lectin and concentration of eggs. Formalin fixed eggs had greater autofluorescence but exhibited the same lectin staining pattern as fresh eggs. The stage of egg development did not influence staining. Eggs from Haemonchus contortus, H. placei, Trichostrongylus colubriformis, T. vitrinus, Ostertagia circumcincta, Nematodirus spathiger and the cestode Monezia expansa were incubated with a panel of fluoroscein isothiocyanate (FITC)-labelled lectins. Lectin binding exhibited a genus specific pattern. Haemonchus spp. stained strongly positive with peanut agglutinin (PNA), and were positive for concanavalin A (ConA), Ricinus communis agglutinin (RCA) and Maclura pomifera lectin (MPA). Trichostrongylus spp. were PNA-, ConA-, RCA- and strongly MPA+. O. circumcincta were weakly positive for PNA, MPA, ConA and negative for RCA. N. spathiger were weakly positive for the four lectins, and M. expansa were weakly positive for PNA, RCA and MPA and were strongly ConA+. The genus specificity of lectin staining was used to identify the presence of Trichostrongylus and Haemonchus eggs in faeces from sheep with mixed field infections, however correspondence between lectin staining and larval differentiation for identifying a low prevalence of Ostertagia in the field infection was poor. Refinements in methods for rapid egg isolation may improve egg differentiation on the basis of lectin staining, which could be undertaken by flow cytometry or microscopy. PMID- 11934463 TI - The effect of feeding sulla (Hedysarum coronarium) or lucerne (Medicago sativa) on lamb parasite burdens and development of immunity to gastrointestinal nematodes. AB - Two experiments investigated the efficacy of the legume Hedysarum coronarium (sulla), which contains condensed tannins (CTs), for reducing gastrointestinal nematode infections relative to lucerne. Experiment 1 was aimed to show whether the lower faecal egg count (FEC) and larval establishment previously reported in lambs grazing sulla were due to direct effects of the forage on Ostertagia circumcincta and Trichostrongylus colubriformis or were mediated through an enhanced immune response. Experiment 2 evaluated the impact of feeding sulla relative to feeding lucerne (Medicago sativa), before, at, or after larval challenge on subsequent FECs and nematode burdens. In experiment 1, 64 Romney lambs were fed either freshly cut lucerne or sulla (32 lambs per herbage) for the duration of the trial. Within each herbage there were four treatment groups (n=8 per group). Initial levels of immunity were assessed in uninfected (UN) lambs which were maintained parasite-free until challenged with 15,000 O. circumcincta and 15,000 T. colubriformis larvae on day 63, and slaughtered on day 81. The other three treatment groups were trickle-infected with each of 5000 O. circumcincta and 5000 T. colubriformis larvae three times per week from day 1 to 35. Non-steroid infected (CONTROL) and steroid-treated (STER) groups were treated with anthelmintic on day 49 and challenged with 15,000 O. circumcincta and 15,000 T. colubriformis on day 63 and slaughtered on day 81. The STER lambs were given dexamethasone trimethylacetate from day 49 to 81 to determine effects of immunity on parasite infection. From day 35 an establishment group (EST) on each herbage was fed a common pelleted lucerne diet and slaughtered on day 56 to determine nematode establishment during trickle-infection. Diet did not affect FECs but feeding lucerne increased (P<0.05) numbers of T. colubriformis in CONTROL lambs compared to those fed sulla. O. circumcincta numbers were lower (P<0.05) in UN lambs fed sulla than lucerne. The sulla diet was associated with higher (P<0.05) antibody titres against secretory-excretory antigens to adult O. circumcincta and to adult and larval T. colubriformis, so there appeared to be some immunogenic response to the sulla diet but effects upon T. colubriformis numbers were not significant. The second experiment involved 48 Romney lambs grazing conventional pasture which were infected with 10,000 each of O. circumcincta and T. colubriformis larvae either 7 days before, 7 days after, or at the time they commenced grazing either sulla or lucerne. Lambs which grazed sulla had lower (P<0.05) FEC and lower (P<0.05) O. circumcincta burdens than lambs which grazed lucerne but timing of infection had no effect on FEC or worm burdens. T. colubriformis numbers were not affected by treatment or herbage. In conclusion, the sulla diet resulted in lower O. circumcincta numbers compared to lucerne outdoors and some evidence of an immunogenic response was obtained indoors. However, neither the herbage nor the immunogenic response reduced T. colubriformis numbers in either experiment. PMID- 11934464 TI - Persistent activity of topical ivermectin against artificial infestations with Hypoderma lineatum (Diptera: Oestridae). AB - In controlled experiments utilizing artificial infestations, a topical formulation of ivermectin (IVOMEC Pour-On for Cattle) was 100% effective against migrating first instar Hypoderma lineatum for 3 weeks following treatment. Larvae were apparently killed early in the infestation as no significant level of specific antibodies was found in the treated calves. At 4 weeks post-treatment the efficacy remained high at 96%; mortality of larvae in the one calf in which warbles were observed and from which mature third instars were collected, was higher than that seen in the untreated calves, indicating some level of treatment induced effect. No specific antibodies were detected in calves that did not develop palpable warbles. Antibody kinetics in those calves from which viable larvae emerged were typical. The length of activity of this product against early stages of the cattle grub makes it practical to apply treatment up to 3 weeks before the end of fly activity. PMID- 11934465 TI - Seroprevalence of antibodies against Neospora caninum in diagnostic equine serum samples and their possible association with fetal loss. AB - A case-control study of the association between the presence of serum antibodies against Neospora spp. and fetal loss was performed on serum samples submitted to a veterinary diagnostic laboratory in northwestern United States. Control sera were randomly selected from those submitted from healthy horses for routine equine infectious anemia testing required for regulatory health certification. Case sera were randomly selected from those submitted from aborting mares for diagnostic workup. Based on a 1:50 or greater titer on the indirect fluorescent antibody test, 8% of the 160 control sera and 13% of the 140 case sera were titer positive. The association odds ratio of 1.67 fell short of statistical significance (p=0.124). PMID- 11934466 TI - Flea species from dogs in three cities of Chile. AB - The purpose of this study was to establish the pattern of flea infestation in dogs in the cities of Santiago (33 degrees latitude south, 70 degrees longitude west), Concepcion (36 degrees latitude south, 73 degrees longitude west) and Osorno (40 degrees latitude south, 73 degrees longitude west), located in different parts of Chile. The only species of fleas found on dogs from the three cities were Ctenocephalides felis felis (41.8%), Ctenocephalides canis (39.4%) and Pulex irritans (18.8%). Although the three species were found in the three cities, differences regarding their frequencies were detected (p<0.05). C. felis was the predominant species found on dogs in Santiago (80.5%). In Concepcion, their frequencies followed a similar trend. However in Osorno, the most southern city, the predominant species was C. canis (78.7%). PMID- 11934468 TI - Angiogenic and astroglial responses to vascular endothelial growth factor administration in adult rat brain. AB - The effects of exogenous vascular endothelial growth factor (VEGF) on angiogenesis, blood-brain barrier permeability and astroglial proliferation in the adult rat CNS in situ were investigated. Recombinant human VEGF(165) (25 or 50 ng/ml) was delivered for up to 1 week using either intracerebral osmotic minipumps or less traumatic subdural gelatin sponge placement. By 3 days, VEGF delivery caused significantly increased cerebral angiogenesis (25 ng/ml was most effective) in both experimental models when compared to saline controls; VEGF infusion resulted in a 100% increase in an index of vascular proliferation, and gelatin sponge delivery produced a 65% increase. The blood-brain barrier hallmark endothelial glucose transporter-1 was not present in nascent vascular sprouts. Infusion of VEGF produced extensive protein leakage that persisted after saline induced permeability was mostly resolved, while gelatin sponge administration caused milder barrier dysfunction. Administration of the angiogenic factor had unexpected proliferative effects on astroglia in both models, resulting in an 80 85% increase in mitotically active astroglia when compared to controls. Immunohistochemical results and semi-quantitative reverse transcriptase polymerase chain reaction indicated that the VEGF receptors flk-1 and flt-1 were up-regulated in response to the infusion trauma; flt-1 was localized to reactive astroglia, while flk-1 was expressed in vascular endothelium but predominantly in neuronal somata and processes adjacent to the delivery site. mRNA for the VEGF(121), VEGF(165) and VEGF(188) isoforms was also increased after delivery of the recombinant protein. These data show that VEGF application has substantial proliferative effects on CNS endothelium and astroglia and causes up-regulation of its own message. Flt-1 and flk-1 receptor mRNAs and proteins are up-regulated in both vascular and non-vascular cell types following infusion trauma. From these results we suggest that administered VEGF has heretofore unanticipated pleiotrophic effects in the adult CNS. PMID- 11934469 TI - Mild traumatic brain injury induces apoptotic cell death in the cortex that is preceded by decreases in cellular Bcl-2 immunoreactivity. AB - Although mild traumatic brain injury is associated with behavioral dysfunction and histopathological alterations, few studies have assessed the temporal pattern of regional apoptosis following mild brain injury. Anesthetized rats were subjected to mild lateral fluid-percussion brain injury (1.1-1.3 atm), and brains were evaluated for the presence of in situ DNA fragmentation (terminal deoxynucleotidyl transferase-mediated dUTP-biotin nick end-labeling, TUNEL) and morphologic characteristics of apoptotic cell death (nuclear and cytoplasmic condensation, presence of apoptotic bodies). Significant numbers of apoptotic TUNEL(+) cells were observed in the injured parietal cortex and underlying white matter up to 72 h post-injury (P<0.05 compared to sham-injured-injured), with maximal numbers present at 24 h. Apoptosis was confirmed by the presence of 180 200 bp nuclear DNA fragments in tissue homogenates. The appearance of apoptotic TUNEL(+) cells in the injured cortex was preceded by a marked decrease in immunoreactivity for the anti-cell death protein, Bcl-2, as early as 2 h post injury. This decrease in cellular Bcl-2 staining was not accompanied by a concomitant loss of staining for the pro-cell death Bax protein, suggesting that post-traumatic neuronal death in the cortex may be dependent on altered cellular ratios of Bcl-2:Bax. In the hippocampus, no significant increase in apoptotic TUNEL(+) cells was observed compared to sham-injured-injured animals. However, selective neuronal loss was evident in the CA3 region at 24 h post-injury, that was preceded by an overt loss of neuronal Bcl-2 immunoreactivity at 6 h. No changes in either cellular Bcl-2 or Bax expression were observed in the thalamus or white matter at any time post-injury. Taken together from these data, we suggest that apoptosis contributes to cell death in both gray and white matter, and that decreases in cellular Bcl-2 may, in part, be associated with both apoptotic and non-apoptotic cell death following mild brain trauma. PMID- 11934470 TI - Acquisition of a novel behavior induces higher levels of Arc mRNA than does overtrained performance. AB - Arc (also termed activity-regulated cytoskeleton-associated protein or Arg3.1), is an effector immediate early gene whose upregulation has been demonstrated during events of synaptic plasticity. In the present study, the possibility that Arc would be specifically upregulated in rats during the acquisition of a quickly learned behavioral task but not in overtrained animals was investigated. Three groups of rats, pseudotrained, newly trained and overtrained, were examined with respect to Arc expression following training on a simple operant lever-pressing task. Newly trained animals were killed 30 min following the session in which they demonstrated acquisition of the task, and overtrained animals were trained on the same task for 13-14 days and then killed. Relative to base level measures taken 6 h following the session, all three groups demonstrated significant levels of induction of Arc mRNA; however, newly trained animals exhibited heightened induction of Arc mRNA relative to both pseudotrained and overtrained animals. The increased levels of Arc mRNA in newly trained animals were located in the CA1 and CA3 fields of hippocampus, the subiculum, and the anterior cingulate, piriform, infra/prelimbic, perirhinal and entorhinal cortical areas. Additionally, Arc mRNA was expressed differentially across the above anatomic structures in a relative pattern that was the same in all three groups. Finally, levels of Arc mRNA in specific brain regions of newly trained animals correlated negatively with the rate of task acquisition, such that slow learners exhibited higher levels of Arc mRNA than fast learners. From these results we suggest that Arc is upregulated in an experience-dependent manner, with higher levels of induction occurring during the initial stage of learning. Furthermore, the finding of increased Arc levels in slow versus fast learners indicates that Arc expression may be associated with the length of time required to: (1) form new associations or (2) remodel existing connections. These results confirm other reports that Arc is a critical substrate for the synaptic plasticity underlying the acquisition of new behaviors. PMID- 11934472 TI - Delayed NGF infusion fails to reverse axotomy-induced degeneration of basal forebrain cholinergic neurons in adult p75(LNTR)-deficient mice. AB - The p75 low-affinity neurotrophin receptor (p75(LNTR)) appears to have various functions that include enhancing nerve growth factor (NGF)-mediated survival by increasing TrkA (high-affinity NGF receptor) efficiency, and mediating apoptosis by acting as a ligand-regulated pro-apoptotic receptor. Here, we investigated the role of p75(LNTR) for adult cholinergic basal forebrain neurons by comparing neuronal responses to injury in control and p75(LNTR)-deficient mice. In both types of mice, approximately 70% of the cholinergic neurons in the ipsilateral medial septum had lost their markers choline acetyltransferase and tyrosine kinase A by 28 days following unilateral transection of the dorsal septohippocampal pathway (fimbria fornix). A 7-day delayed infusion of NGF that started 28 days after the injury resulted in reversal of choline acetyltransferase expression and cell atrophy in control, but not in p75(LNTR) deficient, mice. This lack of response to delayed NGF treatment in p75(LNTR) deficient mice was most likely not due to cell death, as all of the septohippocampal neurons, labeled with Fluorogold before the lesion, were present at 28 days post-lesion, similar to control mice. p75(LNTR)-deficient cholinergic neurons can respond to NGF as they were protected by NGF infusions that started immediately after the injury. These observations, the fact that lesioned p75(LNTR)-deficient neurons atrophy faster, and that non-lesioned neurons hypertrophy in response to NGF in control but not in p75(LNTR)-deficient mice, suggest that p75(LNTR) is needed for tyrosine kinase A and NGF signaling efficiency.In conclusion, during adulthood p75(LNTR) appears to play a beneficial role in the response of cholinergic neurons to injury, consistent with the proposed role of p75(LNTR) in the enhancement of TrkA signaling and the transport of neurotrophins by these neurons. PMID- 11934471 TI - Acetylcholinesterase knockouts establish central cholinergic pathways and can use butyrylcholinesterase to hydrolyze acetylcholine. AB - Acetylcholinesterase is one of the most prominent constituents of central cholinergic pathways. It terminates the synaptic action of acetylcholine through hydrolysis and yields the choline moiety that is necessary for transmitter recycling. Despite these pivotal relationships, mice nullizygous for acetylcholinesterase established all principal anatomical components of central cholinergic pathways. No compensatory increase in the distribution of butyrylcholinesterase was detected. However, both the wild-type and nullizygous mice showed that butyrylcholinesterase enzyme activity extended to all parts of the brain receiving cholinergic innervation and that it could hydrolyze the acetylcholine surrogate acetylthiocholine. As opposed to acetylcholinesterase which was mostly of neuronal origin, butyrylcholinesterase appeared to be mostly of glial origin. These experiments lead to the unexpected conclusion that acetylcholinesterase is not necessary for the establishment of cholinergic pathways. They also show that butyrylcholinesterase can potentially substitute for acetylcholinesterase and that this enzyme is likely to play a constitutive (rather than just back-up) role in the hydrolysis of acetylcholine in the normal brain. The inhibition of butyrylcholinesterase may therefore provide a desirable feature of cholinergic therapies, including those aimed at treating Alzheimer's disease. PMID- 11934473 TI - Injury to retinal ganglion cells induces expression of the small heat shock protein Hsp27 in the rat visual system. AB - Optic nerve transection results in apoptotic cell death of most adult rat retinal ganglion cells that begins at 4 days and leaves few surviving neurons at 14 days post-injury [Berkelaar et al. (1994) J. Neurosci. 14, 4368-4374]. The small heat shock protein Hsp27 has recently been shown to play a role in sensory neuron survival following peripheral nerve axotomy [Lewis et al. (1999) J. Neurosci. 19, 8945-8953]. To investigate the role of Hsp27 in injured CNS sensory neurons, we have studied the induction and cell-specific expression of Hsp27 in rat retinal ganglion cells 1-28 days after optic nerve transection. Immunohistochemical results indicate that Hsp27 is not present at detectable levels in the ganglion cell layer of control (uninjured) or sham-operated control rats. In contrast, Hsp27 is detected in retinal ganglion cells from 4 to 28 days following axotomy. Furthermore, the percentage of surviving retinal ganglion cells that are Hsp27 positive increased over the same time period. Hsp27 is also detected in glial fibrillary acidic protein-positive astrocytes in the optic layer of the superior colliculus from 4 to 28 days after optic nerve transection. These experiments demonstrate that transection of the optic nerve results in the expression of Hsp27 in three distinct regions of the rat visual system: sensory retinal ganglion cells in the eye, glial cells of the optic tract, and astrocytes in the optic layer of the superior colliculus. Hsp27 may be associated with enhanced survival of a subset of retinal ganglion cells, providing evidence of a protective role for Hsp27 in CNS neuronal injury. PMID- 11934474 TI - Effect of Bcl-2 overexpression on establishment of ipsilateral retinocollicular projection in mice. AB - During perinatal development in rodents, ipsilateral retinofugal projection spreading over the superior colliculus is eventually restricted to the rostromedial region. Since this restriction is accompanied by the apoptotic death of more than half of the retinal ganglion cells (RGCs), cell death is believed to play a major role in the restriction of transient ipsilateral projection from the retina to the superior colliculus. To determine the role of RGC death in the establishment of ipsilateral retinofugal projection, we examined the projection pattern in the superior colliculus and the dorsal lateral geniculate nucleus of transgenic mice overexpressing the human bcl-2 gene, which protects against cell death in the CNS. Retrograde labeling of RGCs showed that the number of ipsilaterally projecting RGCs in adult transgenic mice was approximately twice that in adult wild-type mice, indicating that the naturally occurring death of RGCs was prevented in these mutant mice. However, anterograde labeling of ipsilateral retinofugal pathways revealed that the innervation of retinogeniculate and retinocollicular projections was as restricted in transgenic mice as in wild-type mice. From these results we suggest that restriction of ipsilateral retinofugal projection during development is due to retraction or elimination of excessive terminals rather than to naturally occurring RGC death. PMID- 11934475 TI - Altered dendritic development of cerebellar Purkinje cells in slice cultures from protein kinase Cgamma-deficient mice. AB - Protein kinase C (PKC) is a key molecule for the expression of long-term depression at the parallel fiber-Purkinje cell synapse in the cerebellum, a well known model for synaptic plasticity. We have recently shown that activity of PKC also profoundly affects the dendritic morphology of Purkinje cells in rat cerebellar slice cultures suggesting that synaptic efficacy and dendritic development may be controlled by similar intracellular signalling pathways. Here we have analyzed the role of the gamma-isoform of protein kinase C (PKCgamma), which is strongly and specifically expressed in Purkinje cells, during dendritic development. After pharmacological treatment with PKC modulators, phosphorylation of PKCgamma at serine 660 was altered in cerebellar slices suggesting that a change of PKCgamma activity by these treatments was taking place within the Purkinje cells. In PKCgamma-deficient mice, Purkinje cell dendritic trees were enlarged and had an increased number of branching points compared to wild-type mice indicating a role for the PKCgamma isoform as a negative regulator of dendritic growth and branching. Furthermore, the branching-stimulating effects of the PKC inhibitors 2-[1-(3-dimethylaminopropyl)indol-3-yl]-3-(indol-3 yl)maleimide and Go6976 found in wild-type cultures were abolished in the absence of PKCgamma. In contrast, the strong inhibitory effect on dendritic growth by the PKC activator phorbol-12-myristate-13-acetate (PMA) did not require the presence of the PKCgamma isoform since it was still present in the cultures of PKCgamma deficient mice. Our results clearly demonstrate an involvement of PKCgamma in Purkinje cell dendritic differentiation in cerebellar slice cultures. PMID- 11934476 TI - C1 adrenergic neurons are contacted by presynaptic profiles containing DELTA opioid receptor immunoreactivity. AB - Ligands of the delta-opioid receptor tonically influence sympathetic outflow. Some of the actions of delta-opioid receptor agonists may be mediated through C1 adrenergic neurons in the rostral ventrolateral medulla. The goal of this study was to determine whether C1 adrenergic neurons or their afferents contain delta opioid receptors. Single sections through the rostral ventrolateral medulla were labeled for delta-opioid receptor using the immunoperoxidase method and the epinephrine synthesizing enzyme phenylethanolamine N-methyltransferase (PNMT) using the immunogold method, and examined at the light and electron microscopic level. Few ( approximately 5% of 903) profiles dually labeled for PNMT and delta opioid receptor were detected; most of these were dendrites with diameters < 1.5 microm. delta-Opioid receptor immunoreactivity was affiliated with multivesicular bodies in dually labeled perikarya, whereas delta-opioid receptor immunoperoxidase labeling appeared as isolated clusters within both singly and dually labeled dendrites. The majority ( approximately 83% of 338) of delta opioid receptor-immunoreactive profiles were axons and axon terminals. delta Opioid receptor-immunoreactive terminals averaged 0.75 microm in diameter, contained numerous large dense-core vesicles and usually formed appositions or asymmetric (excitatory-type) synapses with their targets. The majority (>50% of 250) of delta-opioid receptor-immunoreactive axons and axon terminals contacted PNMT-immunoreactive profiles. Most of the contacts formed by delta-opioid receptor-immunoreactive profiles ( approximately 75% of 132) were on single labeled PNMT-immunoreactive dendrites with diameters <1.5 microm. The prominent localization of delta-opioid receptors to dense-core vesicle-rich presynaptic profiles suggests that delta-opioid receptor activation by endogenous or exogenous agonists may modulate neuropeptide release. Furthermore, the presence of delta-opioid receptors on axon terminals that form excitatory-type synapses with PNMT-immunoreactive dendrites suggests that delta-opioid receptor ligands may modulate afferent activity to C1 adrenergic neurons. The observation that some PNMT-immunoreactive neurons contain delta-opioid receptor immunoreactivity associated with multivesicular bodies and other intracellular organelles suggests that some C1 adrenergic neurons may present, endocytose and/or recycle delta opioid receptors. PMID- 11934477 TI - Effects of continuous lumbar intrathecal infusion of leptin in rats on weight regulation. AB - Intracranial leptin alters food consumption and body weight. To systematically characterize the effects of extended continuous spinal intrathecal delivery on such regulation, female rats received continuous lumbar spinal infusion (14 days) through catheters connected to osmotic minipumps of a vehicle or one of several doses of recombinant murine leptin (0.03-10 microg/day). The following observations were made. (1) Leptin resulted in a dose-dependent suppression in body weight and food consumption at doses above 0.3 microg/day. (2) Food consumption was initially reduced. Weight fell for 7 days and then plateaued at a level proportional to dose. (3) The ratio of food consumed to body weight was constant for control animals across the study. Leptin-infused rats slowed the initial fall in weight by increasing food consumption, such that the food to body weight ratio returned to that of control values. Rats were thus regulating food consumption to sustain body weight as defined by leptin dose. (4) On day 14, cisternal cerebrospinal fluid was obtained and leptin measured. Concentrations covaried in a log linear fashion with infusion dose. Body weight and food consumption covaried similarly with cisternal leptin concentrations across dose groups. Our data suggest that steady state infusions of leptin induce a degree of appetite suppression that leads to a steady state level of body weight loss and not simply to a simple block of consumatory behavior. The unexpected potency of the observed effects of intrathecal leptin relative to doses that are required after i.c.v. delivery suggests that at least a portion of the effects of intrathecal leptin may reflect a medullary action. The observed correlation of cisternal leptin levels with the behavioral effects is also consistent with a reliable distribution of the infused leptin to target supraspinal sites. PMID- 11934478 TI - Respiratory pre-motor control of hypoglossal motoneurons in the rat. AB - The goal of this study was to determine the origin and transmission pathway of respiratory drive to hypoglossal motoneurons. First we recorded intracellularly from 28 antidromically activated inspiratory hypoglossal motoneurons (resting membrane potential, -50+/-3 mV), and found that injection of chloride ions had no discernible effect on the shape of their membrane potential trajectories. We concluded that the membrane potential trajectories of these hypoglossal motoneurons were determined primarily by inspiratory excitation. To determine the origin of this excitation we cross-correlated the extracellular discharge of medullary inspiratory neurons, including those in the hypoglossal motor nucleus, with the hypoglossal nerve discharge. We found 27 inspiratory neurons within the hypoglossal motor nucleus that were not antidromically activated from the ipsilateral hypoglossal nerve; their cross-correlograms featured either central peaks (1.7+/-0.2 ms) alone (n=14; 39%), or central peaks (1.3+/-0.2 ms) followed by troughs (1.3+/-0.1 ms) at short latencies (1.1+/-0.4 ms) (n=13; 36%), and suggest that these neurons are hypoglossal interneurons. We recorded from 238 inspiratory neurons throughout the rest of the medulla; the cross-correlograms of 19 neurons (8%), located mostly in the lateral tegmental field, displayed narrow half-amplitude peaks (1.0+/-0.1 ms) at short latencies (0.9+/-0.1 ms), which we interpreted as evidence for monosynaptic excitation of hypoglossal motoneurons.We conclude that the respiratory control of hypoglossal motoneurons originates from inspiratory premotor neurons scattered throughout the lateral tegmental field and interneurons within the hypoglossal motor nucleus. PMID- 11934479 TI - Characterization of acid-sensitive ion channels in freshly isolated rat brain neurons. AB - Transient proton-activated currents induced by rapid shifts of the extracellular pH from 7.4 to < or =6.8 were recorded in different neurons freshly isolated from rat brain (hypoglossal motoneurons, cerebellar Purkinje cells, striatal giant cholinergic interneurons, hippocampal interneurons, CA1 pyramidal neurons and cortical pyramidal neurons) using whole-cell patch clamp technique. Responses of hippocampal CA1 pyramidal neurons were weak (100-300 pA) in contrast to other types of neurons (1-3 nA). Sensitivity of neurons to rapid acidification varied from pH(50) 6.4 in hypoglossal motoneurons to 4.9 in hippocampal interneurons. Proton-activated currents were blocked by amiloride (IC(50) varied from 3.6 to 9.5 microM). Reversal potential of the currents was close to E(Na), indicating that the currents are carried by sodium ions. The data obtained suggest that the proton-activated currents in the neurons studied are mediated by acid-sensitive ion channels. Strong acidification (pH<4) induced biphasic responses in all neuron types: the transient current was followed by a pronounced sustained one. Sustained current was not blocked by amiloride and exhibited low selectivity for sodium and cesium ions. Slow acidification from pH 7.4 to 6.5 did not induce detectable whole-cell currents. At pH 6.5, most of the channels are desensitized and responses to fast pH shifts from this initial level are decreased at least 10 times. This suggests that slow acidification which is well known to accompany some pathological states should rather desensitize than activate acid-sensitive ion channels and depress their function. Our results provide evidence for a widespread and neuron-specific distribution of acid-sensitive ion channels in the brain. The large amplitudes and transient character of currents mediated by these channels suggest that they could contribute to fast neuronal signaling processes. PMID- 11934480 TI - Distribution and intraneuronal trafficking of a novel member of the chromogranin family, NESP55, in the rat peripheral nervous system. AB - NESP55 (neuroendocrine secretory protein of M(r) 55000) is a novel member of the chromogranin family. In the present study, we have investigated the distribution, axonal transport and proteolytic processing of NESP55 in the peripheral nervous system. The amount of NESP55 immunoreactivity in adrenal gland was more than 240 times higher than that in the vas deferens. Double or triple immunostaining demonstrated that NESP55 immunoreactivity was highly co-localized with tyrosine hydroxylase immunoreactivity in bundles of thin axons and postganglionic sympathetic neurons; that NESP55 immunoreactivity also co-existed with vesicular acetylcholine transporter immunoreactivity in large-sized axons in sciatic nerves, and that NESP55 immunoreactivity overlapped with calcitonin gene-related peptide immunoreactivity in some large-sized axons, but NESP55 immunoreactivity was not detected in sensory neurons. Strong NESP55 immunoreactivity was found in cell bodies and axons, but it was not detectable in any terminal region by immunohistochemistry. In crush-operated sciatic nerves, NESP55 immunoreactivity could be found as early as 1 h after operation, and accumulated amounts increased substantially with time. However, NESP55 immunoreactivity was only observed in axons proximal to the crush, but none or very little distal to the crush, which was consistent with the data from radioimmunoassay. Finally, extracts of the normal and crushed sciatic nerve and vas deferens were subjected to high performance liquid chromatography followed by radioimmunoassay. The results indicate that NESP55 is processed slowly to small peptides (GAIPIRRH) during axonal transport. NESP55 immunoreactivity was only detected in axons proximal to the crush. The data in the present study indicate that NESP55 immunoreactivity is widely distributed in adrenergic, cholinergic, and peptidergic neurons, but not in sensory neurons, and that this peptide is anterogradely, but not retrogradely, transported with fast axonal transport and slowly processed to smaller peptides during axonal transport in the peripheral nervous system. PMID- 11934481 TI - Cannabinoid 1 receptors are expressed by nerve growth factor- and glial cell derived neurotrophic factor-responsive primary sensory neurones. AB - Expression of the cannabinoid 1 (CB1) receptor and its regulation were studied in the different nociceptive and non-nociceptive sub-populations of cultured primary sensory neurones of adult rats. Bandairaea simplicifolia isolectin B4 (IB4) binding and calcitonin gene-related peptide (CGRP) immunostaining were used to distinguish between the glial cell-derived neurotrophic factor (GDNF)- and nerve growth factor (NGF)-responsive nociceptive and the non-nociceptive primary sensory neurones while a specific CB1 receptor antibody was used to study the expression of the CB1 receptor protein. About half of the total number of primary sensory neurones (47+/-3.2%) cultured for 1 day in the presence of both neurotrophic factors (50 ng/ml each) showed CB1 receptor-like immunostaining, whereas 21.8+/-3.3% and 32.7+/-5.6% of the neurones showed CGRP-like immunopositivity and IB4 binding, respectively. A proportion of the CB1 receptor like immunopositive neurones was immunostained for CGRP (31.7+/-5%) and IB4 (48.2%+/-7.5), with a minimal (1%) co-expression of CGRP and IB4 binding. About a fifth of the CB1 receptor-like immunopositive neurones did not show either CGRP like immunostaining or IB4 binding. To find out whether CB1 receptor expression in nociceptive primary sensory neurones is regulated by GDNF or NGF, cultures were grown in the presence or absence of the neurotrophic factors for 7 days. Vanilloid receptor 1 (VR1) immunostaining was used as a control marker to monitor the effect of the neurotrophins. In cultures maintained in the presence of both factors (50 ng/ml each) 51+/-2.6% and 42.4+/-1.2% of the cells showed CB1 receptor-like and VR1-like immunostaining, respectively. In cultures grown for 7 days in the absence of either of the neurotrophic factors the relative number of VR1-like immunopositive cells decreased to 13.4+/-2.7%, whereas the relative number of CB1 receptor-like immunopositive neurones was unchanged (50.6+/-1.1%). Our data suggest that the CB1 receptor is expressed in all of the three major sub populations of primary sensory neurones and that the CB1 receptor expression is not regulated by either NGF or GDNF. PMID- 11934482 TI - Pre- and postsynaptic localizations of the CB1 cannabinoid receptor in the dorsal horn of the rat spinal cord. AB - Several lines of evidence show that endogenous and exogenous cannabinoids modulate pain transmission at the spinal level through specific cannabinoid-1 (CB1) receptors. Since anatomical data concerning spinal CB1 receptors are rather contradictory, we studied the cellular and subcellular localizations of the CB1 receptors by immunocytochemistry. Results show a dual pre- and postsynaptic localization of CB1 receptors. Presynaptic receptors are evidenced by the labeling of (1) heterogeneous dorsal root ganglion neurons and (2) axons of Lissauer's tract. Postsynaptic receptors are shown by the labeling of numerous interneurons in the outer part of lamina II. Double immunolabelings show that lamina II outer CB1 neurons, probably islet cells, may also contain GABA or nitric oxide synthase. Numerous CB1-containing neurons in lamina X are also immunostained with anti-nitric oxide synthase (NOS) antibody. Under the electron microscope, CB1 immunoreactivity is exclusively localized postsynaptically in both somatic and dendritic compartments. The absence of labeling on primary afferent axon terminals is discussed and compared to the absence of labeling on terminals or vesicle-containing dendrites of islet cells, where a presynaptic localization was expected according to data of the literature. PMID- 11934483 TI - Pathological changes of isolated spinal cord axons in response to mechanical stretch. AB - White matter strips extracted from adult guinea-pig spinal cords were maintained in vitro and studied physiologically using a double sucrose gap technique and anatomically using a horseradish peroxidase assay. The amplitude of compound action potentials was monitored continuously before, during, and after elongation. Three types of conduction blocks resulting from stretch injury were identified: an immediate, spontaneously reversible component, which may result from a transient increase in membrane permeability and consequent disturbance of ionic distribution; a second component that was irreversible within 30-60 min of recording, perhaps resulting from profound axolemmal disruption; and a third component, which may be due to perturbation of the myelin sheath, that was reversible with application of 100 microM of the potassium channel blocker, 4 aminopyridine. The intensity of the conduction deficits correlated with the extent of initial stretch over a full range of severity. Stimulus-response data indicate that mechanical damage to axons in stretch was evenly distributed across the caliber spectrum. Morphological examinations revealed that a small portion of axons exhibited membrane damage at 2 min following stretch and appeared to be largely sealed at 30 min after injury. Further, in the entire length of the cord strip subjected to stretch, axons closer to the surface were found to be more likely to suffer membrane damage, which distinguished stretch injury from compression injury. In summary, we have developed an in vitro model of axonal stretch that provides the ability to monitor changes in the properties of central myelinated axons following stretch injury in the absence of pathological variables related to vascular damage. This initial investigation found no evidence of secondary deterioration of axons in the first 30 min after stretch in vitro, although there was evidence of both transient and lasting physiological and anatomical damage to axons and their myelin sheaths. PMID- 11934485 TI - Diversity and phylogenetic analysis of bacteria in the mucosa of chicken ceca and comparison with bacteria in the cecal lumen. AB - We reported the first attempt to describe mucosa-associated bacterial populations in the chicken ceca by molecular analysis of 16S rRNA genes. Bacteria in the mucosa were highly diverse but mainly Gram-positive with low G+C. Fusobacterium prausnitzii and butyrate-producing bacteria comprised the largest groups among 116 cloned sequences. Twenty five percent of the clones had less than 95% homology to database sequences. Many sequences were related to those of uncultured bacteria identified in human feces or the bovine rumen. Terminal restriction fragment length polymorphism (T-RFLP) analysis revealed some differences between bacterial populations present in the mucosa and lumen of ceca. Greater resolution of bacterial population was obtained using a culture independent approach rather than a culture-based approach. PMID- 11934484 TI - Peptidergic and nitrergic inhibitory neurotransmissions in the hamster jejunum: regulation of vasoactive intestinal peptide release by nitric oxide. AB - Regulation of vasoactive intestinal peptide (VIP) release by nitric oxide (NO) was investigated in the hamster jejunum. Electrical field stimulation and applied NO (3-100 microM) evoked biphasic hyperpolarizations consisting of an initial transient hyperpolarizing component followed by a second more slowly developing component (late component). The NO synthase inhibitor N(G)-nitro-L-arginine methyl ester (200 microM) abolished the biphasic inhibitory junction potential evoked by electrical field stimulation. The NO scavenger oxyhemoglobin (50 microM) and the guanylate cyclase inhibitor 1H-[1,2,4]oxadiazolo-[4,3 a]quinoxalin-1-one (ODQ; 10 microM) abolished both components of the inhibitory junction potentials and the NO-induced hyperpolarizations. VIP(6-28) (1 microM), which abolished VIP (3 microM)-induced hyperpolarizations, also inhibited the late components of the inhibitory junction potentials and the NO-induced hyperpolarizations. ODQ inhibited VIP release and cAMP production by electrical field stimulation and NO application. N(6)-2,0-Dibutyryladenosine 3',5'-cyclic monophosphate (0.1-3 mM) caused a membrane hyperpolarization. These results suggest that NO may stimulate VIP release from enteric nerves in the hamster jejunum. In addition, we propose that NO and NO-stimulated VIP contribute to the early and late components of the inhibitory junction potentials, respectively, in the circular smooth muscle cells of the hamster jejunum. PMID- 11934486 TI - A heteroduplex method for detection of targeted sub-populations of bacterial communities. AB - We describe a simple method, based on heteroduplex mobility analysis of 16S rDNA fragments, for targeted detection of sub-populations of bacteria within diverse microbial communities. A small (ca. 200 bp) polymorphic fragment of the bacterial 16S rRNA gene was amplified from sample DNA using universal primers. Sample products were hybridised with a fluorescently labelled fragment amplified from a selected 'reporter' organism representing the target group. The resulting products were resolved and the labelled heteroduplex pairs detected on non denaturing gels using automated DNA detection technology. A model, based on analysis of samples with known 16S rDNA sequences, demonstrates that heteroduplex mobility is inversely correlated with genetic distance and that beyond 26% genetic difference, heteroduplex products are not detected. The utility of the method was tested by field studies in which stream biofilms could be characterised by heteroduplex profiles generated with heterotrophic and autotrophic reporter organisms representing target groups. PMID- 11934487 TI - Use of the gyrB gene for the identification of Pandoraea species. AB - The recently described genus Pandoraea consists of five named species and four unnamed genomospecies, several of which have been identified in clinical specimens including respiratory secretions from persons with cystic fibrosis. We investigated whether it is possible to distinguish species of the genus Pandoraea by means of restriction fragment length polymorphism (RFLP) analysis and direct sequencing of the gyrB gene. Sixty-seven Pandoraea isolates were included. Species-specific RFLP patterns were obtained following digestion of the PCR amplified gyrB gene with MspI. Specificity of RFLP groupings was confirmed by direct sequencing of several representative isolates. Our results indicate that RFLP analysis and sequencing of the gyrB gene are useful for the identification of Pandoraea species. We also found that further taxonomic studies within the beta-Proteobacteria using the gyrB gene would benefit from the development of additional primers allowing more efficient amplification of the gyrB gene. Our data also indicate that the taxonomic status of Pandoraea genomospecies 2 should be reinvestigated. PMID- 11934488 TI - The flavonoid galangin inhibits the L1 metallo-beta-lactamase from Stenotrophomonas maltophilia. AB - The flavonoid galangin inhibits the partially purified metallo-beta-lactamase from Stenotrophomonas maltophilia. The effect was not reversed by the addition of ZnCl(2) suggesting that the inhibitory effect is not related to metal chelation. The flavonoid quercetin also has some inhibitory effect against the enzyme. Using the crystal structure of the enzyme, a molecular modelling study predicts a possible orientation of galangin at the active site of the enzyme. PMID- 11934489 TI - A linear plasmid temperature-sensitive for replication in Streptomyces hygroscopicus 10-22. AB - Streptomyces hygroscopicus 10-22 harbors a conjugative, autonomously replicating linear plasmid pHZ6 of ca. 70 kb, which shows no obvious homology with chromosomal DNA and is temperature-sensitive for replication, being stable in the host at 28 degrees C but easily lost at 37 degrees C. On a lawn of the wild-type S. hygroscopicus 10-22 cured of pHZ6, pHZ6 elicit pocks. Temperature sensitivity seemed to be a unique property for pHZ6 among six linear plasmids tested, including the well-known linear plasmids SLP2 in Streptomyces lividans 1326 and SCP1 in Streptomyces coelicolor A3(2). The distinct identity of pHZ6 from previously identified pHZ1-pHZ5 was demonstrated by the profile of relevant plasmids in six well-defined strains originated from S. hygroscopicus 10-22. PMID- 11934490 TI - Biological effects of DNA damage in the hyperthermophilic archaeon Sulfolobus acidocaldarius. AB - To investigate the generality of efficient double-strand break repair and damage induced mutagenesis in hyperthermophilic archaea, we systematically measured the effects of five DNA-damaging agents on Sulfolobus acidocaldarius and compared the results to those obtained for Escherichia coli under corresponding conditions. The observed lethality of gamma-radiation was very similar for S. acidocaldarius and E. coli, arguing against unusually efficient double-strand break repair in S. acidocaldarius. In addition, DNA-strand-breaking agents (gamma-radiation or bleomycin), as well as DNA-cross-linking agents (mechlorethamine, butadiene diepoxide or cisplatin) stimulated forward mutation, reverse mutation, and formation of recombinants via conjugation in Sulfolobus cells. Although two of the five DNA-damaging agents failed to revert the E. coli auxotrophs under these conditions, all five reverted S. acidocaldarius auxotrophs. PMID- 11934491 TI - Binding of bovine lactoferrin to Streptococcus dysgalactiae subsp. dysgalactiae isolated from cows with mastitis. AB - Three strains of Streptococcus dysgalactiae subsp. dysgalactiae (UT516, UT519, ATCC 27957) were used to determine if bovine lactoferrin (Lf) binds to bacterial cells by biotin avidin binding assay (BABA), enzyme-linked immunosorbent assay (ELISA), and binding inhibition assay. Binding assays revealed that all strains of S. dysgalactiae subsp. dysgalactiae (S. dysgalactiae) evaluated in this study bound to Lf. However, differences in Lf binding capability among strains and between methods used were detected. Binding of Lf was not inhibited by transferrin (Tf) and Lf moiety molecules (mannose, galactose, and lactose) but by Lf. This study demonstrates that S. dysgalactiae bound to bovine Lf in a specific manner. PMID- 11934492 TI - Identification and role in virulence of putative iron acquisition genes from Corynebacterium pseudotuberculosis. AB - Four genes, fagA, B, C and D, encoding products with 32-47% identity to proteins involved in bacterial iron uptake systems, were identified immediately downstream of the Corynebacterium pseudotuberculosis phospholipase D gene. beta Galactosidase assays on a C. pseudotuberculosis strain carrying a fagA-lacZ fusion indicated that the putative fagABC operon was poorly expressed in iron rich media. However, similar experiments in iron-limited media resulted in an approximately three-fold increase in beta-galactosidase activity, suggesting that this operon is regulated by iron in vitro. Although no defect in iron utilization could be determined for a C. pseudotuberculosis fagB(C) mutant in vitro, this mutant showed reduced virulence compared to wild-type in a goat model of caseous lymphadenitis. Thus, expression of the fag genes in the host appears to contribute to virulence. PMID- 11934493 TI - Pseudomurein endoisopeptidases PeiW and PeiP, two moderately related members of a novel family of proteases produced in Methanothermobacter strains. AB - Sequence comparison of pseudomurein endoisopeptidases PeiW encoded by the defective prophage PsiM100 of Methanothermobacter wolfeii, and PeiP encoded by phage PsiM2 of Methanothermobacter marburgensis, revealed that the two enzymes share only limited similarity. Their amino acid sequences comprise an N-terminal domain characterized by the presence of direct repeats and a C-terminal domain with a catalytic triad C-H-D as in thiol proteases and animal transglutaminases. Both PeiW and PeiP catalyze the in vitro lysis of M. marburgensis cells under reducing conditions and exhibit characteristics of metal-activated peptidases. Optimal temperature and pH were determined to be 63 degrees C and 6.4 for His tagged PeiP and 71 degrees C and 6.4 for His-tagged PeiW, respectively. Database search results suggest that PeiW and PeiP are the first two experimentally identified members of a novel family of proteases in a superfamily of archaeal, bacterial, and eukaryotic protein homologs of animal transglutaminases. PMID- 11934494 TI - FlaK of the archaeon Methanococcus maripaludis possesses preflagellin peptidase activity. AB - Archaeal flagellins are initially synthesized as preflagellins with a short, positively charged leader peptide, which is cleaved prior to the incorporation of the mature flagellins into the filament. While preflagellin peptidase activity had previously been detected in methanogen membranes, the enzyme responsible for this activity had not been identified. We show here that FlaK of Methanococcus maripaludis has preflagellin peptidase activity. In an in vitro preflagellin peptidase assay, Escherichia coli membranes overexpressing Methanococcus voltae preflagellin FlaB2 (as substrate) were combined with E. coli membranes overexpressing M. maripaludis FlaK (as enzyme). Cleavage of the preflagellin was demonstrated by immunoblotting using antibody to FlaB2 and detection of a faster migrating cross-reactive species. This activity required detergent in the assay, and was not detected in membranes previously heated to 95 degrees C. This is the first reported identification of the preflagellin peptidase, and aside from the flagellins, this is the first assignment of function to a gene involved in archaeal flagellation. PMID- 11934495 TI - A major chemotaxis gene cluster in Azospirillum brasilense and relationships between chemotaxis operons in alpha-proteobacteria. AB - Azospirillum brasilense shows chemotaxis to a variety of nutrients and oxygen. Genes encoding the central signal transduction pathway in chemotaxis were identified by phenotypic complementation of generally non-chemotactic mutants. Sequencing of a DNA fragment, which complemented two different mutants, revealed a region of five open reading frames translated in one direction and encoding homologs of known genes comprising excitation and adaptation pathways for chemotaxis in other bacterial species. The major chemotaxis gene cluster appears to be essential for all known behavioral responses that direct swimming motility in A. brasilense. Phylogenetic and genomic analysis revealed three groups of chemotaxis operons in alpha-proteobacterial species and assigned the A. brasilense operon to one of them. Interestingly, operons that are shown to be major regulators of behavior in several alpha-proteobacterial species are not orthologous. PMID- 11934496 TI - Genetic and restriction analysis of the 16S-23S rDNA internal transcribed spacer regions of the acetic acid bacteria. AB - The 16S-23S rDNA internal transcribed spacer regions of the acetic acid bacteria were sequenced and evaluated for molecular identification of these bacteria. All the sequenced spacers contained genes for tRNA(Ile) and tRNA(Ala), and the antitermination element. The sequences revealed 56.8-78.3% similarity. By PCR amplification of the spacers from 57 strains of acetic acid bacteria, single products of similar sizes were produced. Digestion of the spacers by HaeIII and HpaII restriction enzymes resulted in 12 distinct groups of restriction types. All the restriction profiles obtained after analysis of microbial populations from vinegar matched one of the 12 groups. PMID- 11934497 TI - Specificity of a heme-assimilating system of Vibrio vulnificus to synthetic heme compounds. AB - Vibrio vulnificus strain L-180, a clinical isolate, can obtain iron from a synthetic heme, iron-tetra(4-sulfonatophenyl)porphyrin (Fe-TPPS), as well as from a natural heme, protoheme. This assimilation of iron bound to TPPS was demonstrated to be a common property of V. vulnificus by testing a total of 27 strains isolated from both clinical and environmental sources. Strain L-180 could also utilize Fe-TCPP, but not Fe-TMPyP, as a sole iron source. TPPS or its complex with a metal ion reduced bacterial multiplication in the broth containing a minimum dose of Fe-TPPS. When inoculated into human serum supplemented with Fe TCPP, L-180 could grow only in the presence of a protease from the same bacterium. In both TPPS and TCPP, each side chain of a porphyrin ring has a negative charge. Therefore, this negative charge may be important for interaction with an outer membrane receptor involving in a heme-assimilating system of V. vulnificus. PMID- 11934498 TI - Environmental investigation of potentially pathogenic Vibrio parahaemolyticus in the Seto-Inland Sea, Japan. AB - Seawater and organic material (live and/or dead matter deposited on any substratum submersed in seawater) were collected during the cool weather season from a coast of the Seto-Inland Sea, Japan, and analyzed to determine Vibrio parahaemolyticus densities and the occurrence of pathogenic strains, defined as those possessing tdh and/or trh genes by the polymerase chain reaction (PCR), using isolated DNA from enrichment culture of the samples. About 95% of the samples were positive for V. parahaemolyticus (with densities of 3 to >1400 cells per 100 ml water or 10 g organic samples) by the most-probable-number (MPN)-PCR technique with species-specific toxR primers, but only 40% were positive by the conventional MPN-culture technique (with densities ranging from 3 to 240 cells per 100 ml water or 10 g organics). Furthermore, the tdh and trh genes were positive in 55% and 20% of samples, respectively, by the MPN-PCR technique. No tdh and trh gene-positive strains were isolated by the conventional MPN-culture procedure. The difference in detection between the MPN-culture and the MPN-PCR techniques appeared to be significant and may be attributed to different detection sensitivities and other factors. PMID- 11934499 TI - Expression of biopterin transporter (BT1) protein in Leishmania. AB - The present work focuses on the growth phase regulated expression of biopterin transporter gene (BT1) from the LD1 locus on chromosome 35 of Leishmania donovani. Antiserum against recombinant BT1 detected a polypeptide of 45 kDa of equal intensity at lag, log and stationary phases of promastigote growth, both in L. donovani strain LSB-7.1 (MHOM/BL/67/ITMAP263), and strain LSB-146.1 (HOM/IR/95/X81), a natural isolate from Isfehan, Iran that caused cutaneous leishmaniasis. However, in both these strains an additional polypeptide of higher molecular mass (50 kDa) was also observed during lag phase only. In addition, polypeptides of 40, 20, 18 and 16 kDa were seen only during the lag and log phases of both strains. Analysis of L. donovani single, double and triple (null) BT1 knockout mutants confirmed that the 45-kDa polypeptide was the BT1 gene product, as it was absent in the null mutant. These results indicate that 45-kDa BT1 protein in Leishmania is consistently and constitutively expressed in all the growth stages of the parasite. PMID- 11934500 TI - Chromosomal analysis of group B streptococcal clinical strains; bac gene-positive strains are genetically homogenous. AB - A collection of 45 epidemiologically unrelated Streptococcus agalactiae strains (group B Streptococcus, GBS), belonging to different serotypes, isolated from pregnant women in China and Russia was studied. Strains were characterized by pulsed-field gel electrophoresis (PFGE) employing hybridization with nine genes potentially involved in virulence. Molecular sizes of GBS genomes varied from 2030 to 2290 kb. Location of the genes under study bac, bca, glnA, scpB, cyl, hylB, lmb, scaA and cfb on the GBS genomes was found to be conserved irrelevant to the serotype. Potential virulence genes scpB, hylB, lmb were located on a 91 kb SmaI fragment that is equal to 4.5% of total genome. Ribotyping of the strains under study revealed three different HindIII, nine EcoRI and 12 PvuII ribotypes among 45 strains. A strong correlation between the PvuII ribotype and the presence of the bac gene was observed, with 21 of 22 bac-positive strains belonging to the same PvuII ribotype P1. PFGE patterns of bac-positive strains were also similar. The possibility of close genetic relatedness of all bac positive strains is discussed. PMID- 11934501 TI - The LcnC homologue cannot replace LctT in lacticin 481 export. AB - Lacticin 481 is produced by Lactococcus lactis subsp. lactis and belongs to subgroup AII of the lanthionine-containing bacteriocins. The putative homodimeric LctT involved in lacticin 481 production shares significant similarities with the 'LcnC' protein encoded by 'lcnC', located on the chromosome of the lactic acid bacterium, L. lactis IL1403. LctT and 'LcnC' belong to the recently defined family of AMS (ABC transporter maturation and secretion) proteins. Inactivation of the 'lcnC' gene demonstrates that it is not responsible for the weak lacticin 481 production observed in a strain expressing only the precursor peptide LctA, and the modification enzyme LctM. This result indicates that the two AMS proteins, 'LcnC' and LctT, are not interchangeable in the machinery of processing/export of lacticin 481. PMID- 11934502 TI - Investigation of the yvgW Bacillus subtilis chromosomal gene involved in Cd(2+) ion resistance. AB - Analysis of the complete genome sequence of Bacillus subtilis has identified the gene yvgW encoding a protein of 703 amino acids with sequence similarity to the cadmium resistance determinant CadA from the Staphylococcus aureus plasmid pI258. Deletion of yvgW (designated cadA) resulted in increased sensitivity of the strain to cadmium. The cadA gene is expressed from its own promoter, and its expression is induced by cadmium. Northern hybridization analysis showed that cadmium induces the synthesis of a 2.2-kb cadA transcript. These results indicate that cadA is the chromosomal determinant to cadmium resistance in B. subtilis. PMID- 11934503 TI - Nitrite reduction in Paracoccus sp. is affected by a novel plasmid pYR1. AB - Two relatively low-copy plasmids of 9 and 16 kb were found to comprise the extrachromosomal DNA of a Paracoccus strain. Reduction of nitrate by plasmid cured cells resulted in a significant intermediate nitrite accumulation as compared to wild-type cells. By examining nitrate reduction by transformants containing one of the two plasmids, it was found that nitrite accumulation was influenced by the 9.0-kb plasmid, designated as pYR1. Subcloning analysis showed that a 1.8-kb fragment of this plasmid affected nitrite accumulation. Sequence analysis of this fragment revealed the presence of five open reading frames. One of the six deduced proteins showed a strong homology to ABC transporters. PMID- 11934505 TI - Haemophilus influenzae and Vibrio cholerae genes for mutH are able to fully complement a mutH defect in Escherichia coli. AB - MutH, MutL and MutS are essential components of the mismatch repair system in Escherichia coli. Whereas mutS and mutL genes are found in most organisms, the mutH gene is limited to some proteobacteria. We show here that the cloned genes of MutH from Vibrio cholerae and Haemophilus influenzae are able to fully complement a mutH defect in E. coli. Moreover, the purified proteins were shown to be dam methylation sensitive endonucleases, which can be activated by the E. coli MutL protein. These results allow to narrow down regions that are important for the interaction of MutH with MutL. PMID- 11934504 TI - Modification of aklavinone and aclacinomycins in vitro and in vivo by rhodomycin biosynthesis gene products. AB - The rdm genes B, C and E from Streptomyces purpurascens encode enzymes that tailor aklavinone and aclacinomycins. We report that in addition to hydroxylation of aklavinone to epsilon-rhodomycinone, RdmE (aklavinone-11-hydroxylase) hydroxylated 11-deoxy-beta-rhodomycinone to beta-rhodomycinone both in vivo and in vitro. 15-Demethoxyaklavinone and decarbomethoxyaklavinone did not serve as substrates. RdmC (aclacinomycin methyl esterase) converted aclacinomycin T (AcmT) to 15-demethoxyaclacinomycin T, which was in turn converted to 10 decarbomethoxyaclacinomycin T and then to rhodomycin B by RdmB (aclacinomycin-10 hydroxylase). RdmC and RdmB were most active on AcmT, the one-sugar derivative, with their activity decreasing by 70-90% on two- and three-sugar aclacinomycins. Aclacinomycin A competitively inhibited the AcmT modifications at C-10. The results presented here suggest that in vivo the modifications at C-10 take place principally after addition of the first sugar. PMID- 11934506 TI - Identification and characterization of toxigenic Bacillus cereus isolates responsible for two food-poisoning outbreaks. AB - The epidemiology of Bacillus cereus strains responsible for food poisoning is scantly known, mostly because the genotypic and toxigenic properties of the B. cereus strains isolated during food-poisoning outbreaks have been never catalogued. The occurrence of two simultaneous food-poisoning outbreaks gave us the opportunity to wonder whether (i) the identity of individual strains isolated from clinical, environmental, and food samples could be established by random amplified polymorphic DNA (RAPD)-PCR and multiplex RAPD-PCR, and (ii) the toxigenic potential of the isolates could be determined by testing their ability to secrete hemolysin BL, phosphatidylcholine-specific phospholipase C, and cereulide, as well as by determining the presence of the genes encoding enterotoxins NHE, T, and FM/S, cytotoxin K, sphingomyelinase, and phosphatidylinositol-specific phospholipase C. This is the first report demonstrating that the combination of several phenotypic and genotypic traits provides a powerful tool for tracing the source of infection of toxigenic B. cereus strains relevant for epidemiological survey. PMID- 11934507 TI - ITS region of Pythium canariense sp. nov., its morphology and its interaction with Botrytis cinerea. AB - A new species Pythium canariense (CI-07), isolated from soil samples taken in the San Nicolas region of the Canary islands (Gran Canaria, Spain), is being described here. This species is characterised by its spherical to pyriform, intercalary to catenulate sporangia, smooth-walled terminal oogonia supplied with monoclinous and diclinous antheridia which at times are branched and wrap around the female gametangia. The fungus has an antagonistic effect on Botrytis cinerea, the grey mould fungus. Morphological features are being given here together with the sequence of the complete internal transcribed spacer region of the nuclear ribosomal DNA of the fungus, its comparison with related species, and some aspects of its antagonistic behaviour with B. cinerea. PMID- 11934508 TI - Mutational analysis of the bglH catabolite-responsive element (cre) in Lactobacillus plantarum. AB - Catabolite repression of the Lactobacillus plantarum bglH gene is mediated by the catabolite control protein A (CcpA). Here we show that the binding site for the protein is a catabolite-responsive element (cre) sequence overlapping the start site of transcription. Two single and one double base substitutions in the cre sequence caused derepression of a plasmid-borne bglH-cat transcriptional fusion in L. plantarum cells grown on glucose. Analysis of the single mutations indicates that the G and C nucleotide residues in positions 2 and 13, respectively, of the 14-bp cre sequence are required for catabolite repression. PMID- 11934509 TI - Sleep apnoea in infancy and childhood. Considering two possible causes: obstruction and neuromuscular disorders. AB - Two aspects of sleep apnoea in infancy and childhood are considered. First of all the obstructive sleep apnoea syndrome is reviewed; its causes, and types, and clinical differences depending on the age of the affected patient. Difficulties of diagnosis are discussed, as well as methods used to confirm the presence of the syndrome. Then means of treatment are considered, both medical and surgical. The second part of the paper is concerned with a particular group of children, especially at risk of sleep apnoea; those suffering from neuromuscular disorders as these are likely to be of special interest to paediatric neurologists. These include neuropathies, myopathies such as Duchenne muscular dystrophy and myotonia, and disorders of the neuromuscular junction. PMID- 11934510 TI - Long-term effectiveness and side effects of acetazolamide as an adjunct to other anticonvulsants in the treatment of refractory epilepsies. AB - The long-term effectiveness of acetazolamide (AZA) and its side effects, especially the formation of renal calculi, were investigated in a prospective study when AZA was used as an adjunct to other antiepileptic drugs in the treatment of refractory epilepsies. The subjects comprised 37 patients aged from 1 to 17 years (mean age, 8 years and 1 month) whose seizures were hard to control with the use of two or more drugs among sodium valproate, carbamazepine and clonazepam. Thirty-two of the 37 patients were complicated with mental retardation. A daily dose of 10mg/kg of AZA was first administered and then the dosage was increased up to 20mg/kg based on the clinical response and side effects. The maintenance daily dosage of AZA (12.2+/-4.2mg/kg) produced a steady state plasma concentration of 6.2+/-4.5 microg/ml. Among the 37 patients, complete seizure control for more than 3 years was obtained in four patients. Although there were no significant differences, all of the four patients were classified as having symptomatic localization-related epilepsies. Seizures recurred in five after complete remission for at least 6 months, and six showed >50% decrease in seizure frequency for more than 6 months after the introduction of AZA. Twenty-eight patients, who were taking AZA for 10 months to 14 years (mean, 6 years and 5 months), were examined for the formation of renal calculi. None of them showed evidence of renal calculi. This study reinforces the idea that AZA may be a useful adjunct drug in selected patients with refractory symptomatic localization-related epilepsies. PMID- 11934511 TI - Postoperative development of children after hemispherotomy. AB - We studied the postoperative development of 14 children with cortical dysgenesis who underwent modified functional hemispherectomy (hemispherotomy) at the age of 6 years or younger. At follow-up of 24-72 months (median of 47 months), six (43%) were seizure-free, six achieved>90% reduction, one achieved 50-90% reduction, and one achieved 0-50% reduction in seizure frequency. At the preoperative and final postoperative examinations, mean scores of developmental quotient (DQ) were as follows: 25.5 and 31.9 in total DQ, 26.0 and 33.7 in intellectual DQ, and 27.4 and 22.9 in motor DQ. Children scoring >50 points in preoperative intellectual DQ score obtained near-normal intellectual DQ postoperatively, while, those scoring <10 DQ preoperatively remained at a low developmental level. Among children with DQ scores in the range from 10 to 50, two children who obtained seizure-free outcome and were operated upon in the first 3 years of life achieved marked developmental progress. The present study indicated that high preoperative intellectual DQ and cessation of seizures seem to be associated with better postoperative intellectual development. However, long-term observation of postoperative development and an accumulation of more cases will be needed before we can reach a firm conclusion. PMID- 11934512 TI - Low-voltage activity in EEG during acute phase of encephalitis predicts unfavorable neurological outcome. AB - The purpose of this study was to evaluate the risk factors responsible for the onset of postencephalitic epilepsy during the acute phase of encephalitis. We retrospectively studied 19 patients with acute encephalitis admitted to the Division of Pediatrics at our hospital from January 1989 to December 1998. Nine cases were complicated by postencephalitic epilepsy and 10 were non-complicated cases. Significant risk factors for postencephalitic epilepsy were seizures, more severe impairment of consciousness, abnormalities in cranial computed tomography/magnetic resonance imaging/single photon emission computed tomography examination and low-voltage EEG activity, which was defined as EEG findings showing less than 20 microV of background activity without the appearance of a hump or spindle, during the acute phase of encephalitis. In particular, cases with low-voltage EEG activity were all complicated by intractable epilepsy after less than 1 year. Low-voltage EEG activity during the acute phase predicts an unfavorable neurological outcome, reflecting severe widespread impairment in the whole of the cortex. PMID- 11934513 TI - Epileptic and non-epileptic cerebral palsy: EEG and cranial imaging findings. AB - The aims of the study were to compare the clinical types, electroencephalogram (EEG) and cranial magnetic resonance imaging/computed tomography findings of epileptic and non-epileptic cerebral palsy (CP) patients. Seventy-four patients with CP were evaluated in 2 years. Tetraplegic CP had a higher incidence of epilepsy (60.5%). EEG was confirmed abnormal in epileptic CP as 90.3%, and in non epileptic CP as 39.5%. Focal epileptiform activity, generalized slowing, and multifocal epileptiform activity were significantly frequent in epileptic CP. There were cranial imaging abnormalities of 74.2% in epileptic and 48.8% in non epileptic CP. Although there was not any statistically significant difference between the two groups, epileptic group revealed more structural abnormalities. Further studies concerning a possible risk of epilepsy development and its relations with the EEG and cranial imaging findings are needed in presenting the other risk factors involved and the factors affecting the CP prognosis. PMID- 11934514 TI - Application of delayed extraction matrix-assisted laser desorption ionization time-of-flight mass spectrometry for analysis of sphingolipids in cultured skin fibroblasts from sphingolipidosis patients. AB - Sphingolipidoses are caused by defects of enzymes involved in the hydrolysis of sphingolipids. Using delayed extraction matrix-assisted laser desorption ionization time-of-flight mass spectrometry (DE MALDI-TOF-MS), we analyzed sphingolipids in cultured skin fibroblasts from patients with sphingolipidoses, including: (a) Farber disease (FD, acid ceramidase deficiency); (b) Gaucher disease (GD); (c) Niemann-Pick disease type C (NPDC); and (d) GM1-gangliosidosis (GM1G). Crude lipids were extracted from about 50 mg wet weight of cultured skin fibroblasts. After mild alkaline treatment, a sphingolipid fraction was prepared from the crude lipids and analyzed by DE MALDI-TOF-MS. The results were as follows: (a) in fibroblasts from the FD patient, the ceramide/sphingomyelin and ceramide/monohexosylceramide ratios were both significantly high; (b) in the GD patient, the glucosylceramide/sphingomyelin ratio was increased; on the other hand; (c) in the NPDC patient, the monohexosylceramide/sphingomyelin ratio was within normal range; and (d) in the GM1G patient, no specific data were obtained. Sphingolipids in cultured fibroblasts can be evaluated by DE MALDI-TOF-MS, whereas GM1-ganglioside or its asialo derivatives are not detectable. With this DE MALDI-TOF-MS method, ceramide or monohexosylceramide accumulating in cultured fibroblasts from cases of sphingolipidoses, such as FD and GD, respectively, can be easily detected. PMID- 11934515 TI - Wisconsin card sorting test in children with temporal lobe epilepsy. AB - To search for the origin of frontal lobe dysfunction identified by the Wisconsin card sorting test (WCST) in temporal lobe epilepsy (TLE) patients, we investigated the WCST performance among 19 children with TLE (with hippocampal atrophy (HA group N=12), without structural lesions (NSL group N=7)), 15 patients with frontal lobe epilepsy (FLE group), and age-matched normal controls (N group). The paired verbal association learning test (PVALT) and Benton visual retention test (BVRT) were also performed. HA group and FLE groups achieved significantly fewer categories and demonstrated more perseverative errors on the WCST than NSL and N groups. In addition, category achievement in WCST showed significant inverse correlation to age at the initial status convulsivus in the HA group (P<0.05). The achievement on PVALT and BVRT did not show any significant differences between HA and FLE groups (P>0.05). Thus, the frontal lobe dysfunction in the HA group is found to exist as early as 7 years old, when they seem to have only a short seizure history or to receive a little electrical interference from the temporal lobe focus to the frontal region. These facts would underscore the importance of prefrontal dysfunction persisting from the early insults and only becoming apparent after maturation of the prefrontal region in patients with mesial TLE. PMID- 11934516 TI - Subclinical central pontine myelinolysis following liver transplantation. AB - Central pontine myelinolysis (CPM) is a demyelinating disorder of unknown origin that almost exclusively affects the central portion of the basis pontis, and is one of the fatal neurological complications after liver transplantation. We describe two children with CPM detected incidentally after liver transplantation. To our knowledge, this is the first report of CPM diagnosed antemortem in children who had undergone liver transplantation. In our patients, there were no clinical manifestations associated with CPM. We conclude that, including silent cases such as our patients, CPM may be more prevalent than previously appreciated following liver transplantation. PMID- 11934517 TI - Gelastic seizures and low-grade hypothalamic astrocytoma: a case report. AB - The typical, well recognized childhood epilepsy syndrome caused by hypothalamic hamartoma is characterized by early-onset, stereotyped attacks of uncontrollable laughter, frequent refractory seizures with progressive cognitive deterioration and severe behavioral problems. Here, we report a 17-year-old patient with gelastic phenomenon started in the neonatal period, later on associated with drug resistant polymorphic seizures, intellectual deficit and behavioral disorders, who improved by partial resection of an expected hypothalamic hamartoma that, in turn, resulted to be a hypothalamic low-grade astrocytoma. PMID- 11934518 TI - Acute transverse myelitis after Japanese B encephalitis vaccination in a 4-year old girl. AB - Fourteen days after Japanese B encephalitis (JBE) vaccination, a 4-year-old girl developed the full clinical manifestation of ATM within 24h. She showed acute ascending flaccid paraplegia with sensory disturbance, bladder dysfunction and meningeal sign. Cerebrospinal fluid examination revealed neutrophil pleocytosis and elevated protein level. Magnetic resonance imaging (MRI) showed diffuse swelling of the cervical and lumbar cord with low signal intensity on T1 and high signal intensity on T2-weighted imaging. These findings suggested that she had developed meningo-radiculomyelopathy. Since sequential MRI studies showed prompt reduction of the cord swelling, the high-dose methylprednisolone therapy employed seemed to have been effective for improvement of inflammation. Even with such potent drug treatment, she still has substantial flaccid diplegia and sphincter disorder 1 year later, and so we are convinced that the pathological change of the cord was as severe as in necrotizing myelopathy. Although the pathological process remains unknown, cellular autoimmune mechanism against the JBE vaccination is suspected. PMID- 11934519 TI - Sequential changes of brain CT and MRI after febrile status epilepticus in a 6 year-old girl. AB - Brain CT or MRI occasionally shows transient or permanent changes in the brain after status epilepticus (SE). The mechanism for these changes has not been well elucidated. We performed repeated imaging studies on a patient with febrile SE characterized by right hemiconvulsion. CT showed transient mild edema on both hemispheres immediately after the cessation of SE. The edema improved the next day. But aphasia and right hemiparesis were observed. On day 17, CT revealed edema on left hemisphere and MRI showed a high signal intensity in cortex and subcortical white matter of the left hemisphere on T2-weighted images. Although right hemiparesis and aphasia were improved, severe atrophy of the left hemisphere was noted on CT and MRI. The results suggest that brain edema observed in several days after SE but not edema observed immediately after the cessation of SE is more pathological for the permanent brain damage. Possible mechanisms of the initial brain edema and the second edema preceded severe atrophy in left hemisphere were discussed. PMID- 11934520 TI - Two successful cases of bromide therapy for refractory symptomatic localization related epilepsy. AB - Potassium bromide was tried for two children with daily convulsive focal motor seizures with unconsciousness and focal motor seizure status. The treatment resulted in complete cessation of the attacks. It has been reported that bromide is effective for generalized tonic-clonic seizures and not for complex partial seizures, such as convulsive focal motor seizures with unconsciousness. However, our experiences provide evidence that bromide is one of the useful therapeutic agents for intractable symptomatic localization-related epilepsy. PMID- 11934521 TI - Auditory perception in auditory neuropathy: clinical similarity with auditory verbal agnosia. AB - The precise features of auditory perception in patients with auditory neuropathy have not been well described. In the present study, we examined auditory perception in a patient with auditory neuropathy. The patient was a right-handed 7-year-old boy. His chief complaint was delayed speech and suspected of verbal learning disability. He could talk, read and repeat rather fluently but could not understand fully what was asked. V-IQ, P-IQ and F-IQ of Wechsler Scale for Children III-R were 53, 118 and 81, respectively. Pure tone audiogram was completely normal. His speech discrimination ability was very poor. He could identify environmental sounds with visual matching. He could differentiate intensity difference but not time difference. This phenomenon was reported in patients with hemispheric symptoms. These clinical features are very similar to verbal auditory agnosia. ABR showed no response at 90dBnHL alternating clicks and tone bursts. Click evoked and distortion product otoacoustic emissions (OAE) were normal. Electrocochleogram was also normal. Motor and sensory nerve conduction velocity was completely normal. Pa of MLR and N1 of SVR were present. His diagnosis should be "pure type" of auditory neuropathy or auditory nerve disease. Importance of both ABR and OAE examination should be widely announced and auditory neuropathy must be campaigned stressed to be clinical entity among personnel who take care of children with speech delay. PMID- 11934522 TI - Effects of frequency of treatment with recombinant equine somatotropin on selected biological responses in geldings. AB - Two experiments compared the efficacies of different treatment frequencies for recombinant equine somatotropin (eST). In Experiment 1, five geldings received daily injections of eST at 20 microg/kg of body weight, and five received every other-day injections at 40 microg/kg of body weight, for a total of 30 days. Plasma glucose (P=0.0001), insulin (P=0.0135), and non-esterified fatty acid (NEFA, P=0.0001) concentrations increased, and plasma urea nitrogen (PUN) concentrations decreased (P=0.0001), in both groups, and only minor differences (P<0.05) occurred between the two groups. Insulin-like growth factor-I (IGF-I) concentrations increased (P=0.0001) in both groups over time, and were higher (P<0.05) after day 2 in geldings treated daily. Endogenous somatotropin (ST) response to secretagogue was inhibited (P<0.05) in geldings receiving daily injections relative to those receiving every-other-day injections. In Experiment 2, 16 geldings were allotted to four groups of four. A control group received daily saline injections, and the other three groups received eST at 20 microg/kg of body weight daily as a single injection, two injections (every 12h), or four injections (every 6h), for a total of 14 days. Plasma IGF-I and insulin concentrations increased (P<0.05) in all groups receiving eST, with the responses being proportional to injection frequency. In contrast, PUN concentrations decreased (P<0.05) in all groups equally. In conclusion, the efficacy of daily versus every-other-day injections of eST depends upon the response to be measured, and for IGF-I concentrations, the every-other-day regimen was not acceptable. Injection frequencies greater than once daily were more efficacious for IGF-I and insulin concentrations, but not for PUN concentrations. Thus, the optimum injection regimen for any new application for eST cannot simply be inferred from other biological responses, and will need to be determined empirically. PMID- 11934523 TI - Production and regulation of leptin in bovine mammary epithelial cells. AB - Previously, leptin has been found in human and rodent mammary tissue. The present research was conducted to determine (1) if leptin is produced by bovine mammary epithelial cells and (2) if leptin production in bovine mammary epithelial cells is hormonally regulated. Western blot analysis indicated the presence of leptin in bovine milk, while reverse transcription-polymerase chain reaction (RT-PCR) indicated the presence of leptin mRNA in mammary tissue and cultured bovine mammary epithelial cells (MAC-T cell line). A real time RT-PCR method was developed that allowed quantitative assessment of bovine leptin mRNA over approximately 3 orders of magnitude. Time course studies indicated a rapid increase in leptin mRNA in response to insulin or insulin-like growth factor-I (IGF-I). When normalized against bovine GAPDH as an internal control, 0.5 or 1h treatment with 10 ng/mL insulin gave 39+/-4 and 64+/-2-fold increase in leptin mRNA compared with 0 h control. Leptin mRNA was increased 257+/-9 and 75+/-23 fold by 0.5 or 1h treatment with 10 ng/mL IGF-I. Dose response studies indicated significant increases in leptin mRNA in response to as little as 1 ng/mL insulin or 0.1 ng/mL IGF-I. Maximum increase in leptin mRNA was observed in response to 10 ng/mL insulin and 10 ng/mL IGF-I. These results indicate that production of leptin by bovine mammary epithelial cells can be regulated by factors known to alter mammary function and nutrient partitioning. This suggests that leptin may be an autocrine/paracrine signal in the bovine mammary gland. PMID- 11934524 TI - Feeding different amounts of colostrum or only milk replacer modify receptors of intestinal insulin-like growth factors and insulin in neonatal calves. AB - Colostrum intake influences growth and development of the gastrointestinal tract (GIT) in several species and colostral insulin-like growth factors I and II (IGF I and IGF-II), and insulin are involved in neonatal intestinal tissue growth. We have studied IGF type 1, IGF type 2, and insulin receptors in the intestine of 8 day-old calves fed different amounts of colostrum or only milk replacer. Calves were fed colostrum of the first six milkings on the first 3 days and then milk replacer (GrC(6)) or colostrum only once and then milk replacer (GrC(1)) or they were fed only milk replacer from the beginning, i.e., no colostrum (GrM). Competitive binding studies and ligand blots confirmed the presence of IGF type 1, IGF type 2, and insulin receptors in mucosal cell membranes of duodenum, jejunum, ileum, and colon. The IGF type 1 receptor number in ileum and total intestine in GrC(6) was greater (P<0.05) than in GrC(1) and in GrM, and IGF type 2 receptor number in total intestine was greater (P<0.05) in GrC(6) than in GrM. Insulin binding was best fitted by a model with two binding sites. High affinity insulin receptor numbers in duodenum, ileum, and total intestine were greater (P<0.05) in GrC(6) than in GrM. The data demonstrate that IGF type 1, IGF type 2, and insulin receptors in intestinal mucosa of neonatal calves are influenced by feeding. PMID- 11934525 TI - Cloning and sequencing of the 3'-region of the canine parathyroid hormone-related protein gene and analysis of alternate mRNA splicing in two canine carcinomas. AB - A canine genomic library in Lambda FIX II vector was screened with a 281-base pair canine PTHrP cDNA to the prepro- and coding regions. Two genomic clones were isolated and mapped to the 3'-end of the PTHrP gene by polymerase chain reaction (PCR) amplification of exons in this region. One clone (3.5 kb) was amplified by PCR, partially sequenced, and compared to the human PTHrP gene. Regions were identified with a high degree of homology to exons 6, 7, and 8 of the human PTHrP gene. A polyadenylation site was present 3' to the exon 8-like region. Reverse transcriptase-polymerase chain reaction (RT-PCR) demonstrated that exon 7 of the PTHrP gene was transcribed in two canine carcinomas (SCC 2/88 cells and CAC-8 tumor line) which produce PTHrP. This confirmed that the 3'-region of the canine PTHrP gene is alternately spliced with splicing of exon 6 to exons 7 or 9. Transcription of exon 8 was not demonstrated by RT-PCR and suggests that the exon 8-like region of the dog PTHrP gene is not utilized. The exon 8-like region contained an early stop codon that was not present in exon 8 of the human PTHrP gene. PMID- 11934526 TI - Effect of the preovulatory LH surge on bovine follicular progesterone receptor mRNA expression. AB - A growing body of evidence indicates that intrafollicular progesterone receptor signaling pathways are obligatory for follicle rupture. However, the intrafollicular localization and regulation of progesterone receptor expression during the periovulatory period in cattle are not known. In this study, we determined the effect of the preovulatory gonadotropin surge on localization and expression of progesterone receptor mRNA in bovine periovulatory follicular and luteal tissue. Ovaries containing preovulatory follicles or new corpora lutea (CL) were collected at approximately 0, 6, 12, 18, 24 (preovulatory follicles) and 48 h (CL) after a GnRH-induced LH surge (n=5-8 per timepoint). Expression of progesterone receptor mRNA was detected in periovulatory follicular and luteal tissue at all timepoints examined. Relative levels of progesterone receptor mRNA were dramatically upregulated within 6h after the LH surge compared to all other time points (P<0.0001). In situ hybridization analysis revealed that the significant increase in progesterone receptor mRNA expression was localized to the granulosal layer of preovulatory follicles. Our results indicate that progesterone receptor mRNA expression is upregulated specifically in the granulosal layer of bovine preovulatory follicles following the LH surge. Progesterone receptor signaling pathways may help mediate the effects of the preovulatory LH surge on follicle rupture in cattle. PMID- 11934527 TI - Atenolol: pharmacokinetic/dynamic aspects of comparative developmental toxicity. PMID- 11934528 TI - The teratogenic effect of carbamazepine: a meta-analysis of 1255 exposures. AB - Maternal use of antiepileptic drugs during pregnancy has been associated with an increased risk of major congenital abnormalities in the fetus. Carbamazepine (CBZ) is an antiepileptic drug that was developed and marketed mainly for the treatment of epileptic seizures. Some investigators described an increased rate of major congenital anomalies following treatment with CBZ during pregnancy while others found no such increase. In order to quantify better the risks of exposure to CBZ during pregnancy, we pooled data from prospective studies known to us. We found in prospective studies involving 1255 cases of exposure that CBZ therapy increased the rate of congenital anomalies, mainly neural tube defects, cardiovascular and urinary tract anomalies, and cleft palate. CBZ may also induce a pattern of minor congenital anomalies and developmental retardation, but our study did not address these endpoints. CBZ also appears to reduce gestational age at delivery. A combination of CBZ with other antiepileptic drugs is more teratogenic than CBZ monotherapy. Children born to untreated epileptic women do not appear to have an increased rate of major birth defects. In light of these results, we recommend performing a level 2 ultrasound and fetal echocardiography in women treated with CBZ during pregnancy. PMID- 11934529 TI - Fetal testosterone insufficiency and abnormal proliferation of Leydig cells and gonocytes in rats exposed to di(n-butyl) phthalate. AB - Adult male rats previously exposed on gestation days (GD) 12-21 to di(n-butyl) phthalate (DBP) have reproductive tract malformations, particularly agenesis of the epididymis, decreased sperm production, and Leydig cell hyperplasia and adenomas. Although similar effects are produced by the potent androgen receptor (AR) antagonist flutamide and are indicative of disruption of male sexual differentiation via an antiandrogenic mechanism, DBP is not an AR antagonist. The purpose of the study was to determine whether DBP causes pathologic changes and alterations in androgen status in the testis during the prenatal period of male reproductive tract differentiation. Pregnant CD rats were given corn oil, DBP (500 mg/kg/day), or flutamide (100 mg/kg/day) p.o. on GD 12-21. At GD 16-21, DBP caused hyperplasia of Leydig cells, many of which were 3beta-hydroxysteroid dehydrogenase- and/or AR-positive. Focal areas of hyperplasia had increased numbers of Leydig cells positive for proliferating cell nuclear antigen (PCNA). At GD 21, testis atrophy was apparent, seminiferous cords in DBP-exposed fetuses were enlarged and contained multinucleated gonocytes that, unlike controls, were PCNA-positive. DBP, but not flutamide, markedly decreased testicular testosterone levels at GD 18 and 21. Fewer epididymal ducts and reduced AR staining in some ducts were evident with DBP treatment, whereas decreased overall AR staining was seen with flutamide in the presence of mild Leydig cell hyperplasia. Leydig cell proliferation is likely a compensatory mechanism to increase testicular steroidogenesis triggered by testosterone insufficiency. The overall decrease in androgen concentration is not corrected and results in reproductive tract malformations. The multinuclearity and proliferation of gonocytes suggests an underlying Sertoli cell dysfunction. PMID- 11934530 TI - Effects of nonylphenol, 1,1-dichloro-2,2-bis(p-chlorophenyl)ethylene (p,p'-DDE), and pentachlorophenol on the adult female guinea pig reproductive tract. AB - The guinea pig exhibits cyclic and luteal similarities to the human, a feature not present in other small experimental animals such as rats, mice, or rabbits. Studies were undertaken to investigate the in vivo effects of three persistent environmental xenobiotics (nonylphenol, 1,1-dichloro-2,2-bis(p chlorophenyl)ethylene [p,p'-DDE], and pentachlorophenol) on the microanatomy of the adult female guinea pig reproductive system. The effects brought about by these compounds (40 mg/kg/day) were compared to those caused by the synthetic estrogen diethylstilbestrol (DES; 50 microg/kg/day). Adult female guinea pigs, intact and castrated, were treated with 14 daily subcutaneous (s.c.) doses of one of these agents. The 50% decline in the weight of the tract that occurred following castration, was prevented by administration of nonylphenol, p,p'-DDE, and DES, but not of pentachlorophenol. Nonylphenol produced weak estrogenic stimulation of the tract of intact animals and maintained a relatively normal histologic appearance in castrated animals. Focal mucinous metaplasia of the endometrium, however, was observed in both groups. Treatment of intact and castrated animals with p,p'-DDE resulted in cystic hyperplasia and mucinous metaplasia of the endometrium, hyperplasia of the cervical epithelium, estrogenic stimulation of the vagina, and dilation of the rete ovarii. Treatment of intact or castrated animals with DES resulted in effects that were qualitatively similar to those caused by p,p'-DDE. The appearance of the vaginal epithelium, however, was abnormal and the rete ovarii were less dilated. Pentachlorophenol had minimal effect on the histology of the tract of castrated or intact animals. These data support our hypothesis that some environmental toxicants can substitute for estradiol in regulating the microanatomy of the female reproductive tract. They indicate the potential of these compounds to act as endocrine disrupting agents. PMID- 11934531 TI - Mass spectrometric determination of p-nonylphenol metabolism and disposition following oral administration to Sprague-Dawley rats. AB - Isomers of 4-nonylphenol (NP), which are important industrial compounds and environmental breakdown products from widely used surfactants, have estrogenic activity in vitro and in vivo that has prompted interest in its potential for modulation of endocrine function in humans and wildlife. Mass spectrometry was used to quantify NP and metabolites in serum and endocrine-responsive tissues from dietary exposure in Sprague-Dawley rats. Tissue accumulation of NP aglycone was observed despite the predominance of glucuronidation in blood. Serum toxicokinetics of total NP, measured following gavage administration, showed rapid absorption and elimination (average half-times 0.8 and 3.5 h, respectively). NP was similarly administered by gavage to pregnant dams and total and aglycone NP were measured in dam serum and fetuses to show placental transfer into serum and brain. These data provide a basis for future correlations of biologic effects observed following dietary exposure in rats with those predicted from environmental exposures to humans. PMID- 11934532 TI - Human sperm immobilizing activity of aminophenyl arsenic acid and its N substituted quinazoline, pyrimidine, and purine derivatives: protective effect of glutathione. AB - We examined the potential toxicity of pentavalent organic arsenicals for human sperm. We used computer-assisted sperm analysis to examine the effects of three aminophenyl arsenicals and their nine N-substituted quinazoline, pyrimidine, and purine derivatives on human sperm motility and kinematics in human semen and medium. Among the arsenicals examined, (aminophenylazo)-phenyl arsonic acid and its N-substituted pyrimidine derivative PHI-370 (2-methylthio-4-[(4' aminophenylazo)-phenylarsonic acid] pyrimidine) exhibited rapid sperm immobilizing activity in medium with EC(50) values of 77 and 82 microM, respectively, and t(1/2) of < 3 min. Molecular modeling analysis indicated that sperm-immobilizing organic arsenicals exhibit high dipole moments (>7 Debyes). Sperm immobilizing activity of these arsenicals was completely abrogated in the presence of seminal plasma. Furthermore, coincubation of motile sperm with PHI 370 in the presence of reduced glutathione (GSH) resulted in dose-dependent protection of sperm motility and sperm motion parameters. Coincubation of the arsenical with GSH at a molar ratio of 1:20 resulted in 95% retention of sperm progressive motility. The mean values of the other sperm movement characteristics also showed > 90% protection. These observations suggest that the rapid sperm immobilizing activity of these pentavalent arsenicals may be as a result of direct binding of the arsenical with the sperm thiol components essential for sperm motility as well as induction of oxidative damage by disruption of sperm cell's antioxidant system. Sodium arsanilate and its N-substituted pyrimidine derivative, PHI-370, are useful probes to further evaluate the mechanism of pentavalent arsanilate-induced human sperm dysfunction. PMID- 11934533 TI - Hexachlorobenzene exposure and the proportion of male births in Turkey 1935-1990. AB - Hexachlorobenzene (C(6)Cl(6), HCB) is a chemical that has been associated with significant immediate and long term adverse health effects in humans. It has been associated with both porphyria cutanea tarda and spontaneous abortions among survivors of widespread exposure in the 1950s in southeastern Turkey. HCB binds to the Ah receptor, albeit with lower affinity than dioxin. Dioxin exposure has been reported to lower human secondary sex ratio, putatively through a male mediated effect. We therefore wished to evaluate the impact of the HCB environmental event on the sex ratio of the progeny of the survivors. We undertook an assessment of 1) the effects of HCB exposure on the proportion of male births of individual subjects who had survived, 2) variables that significantly predicted the proportion of males among these individuals, and 3) the trend of the population sex ratio born in Turkey from 1935 to 1990. Women known to have been exposed to HCB in the 1950s did not have offspring with a significantly different sex ratio when compared to control populations. However, subjects reporting exposure at the peak of the episode (1955-57) had a significantly lower lifetime proportion of males than those exposed at a later date. The lifetime reported spontaneous abortion rate of these women also significantly predicted the percent males per subject. The available national data demonstrated a significant reduction in the calculated proportion of males from 1935 to 1970 that stabilized from 1970 to 1990. These data indicate that HCB exposure that was sufficient to induce clinical porphyria cutanea tarda may also have reduced the proportion of males in subjects over their reproductive life span. The HCB episode does not explain the pattern of the national trend from a population perspective. PMID- 11934534 TI - Effects on development of the reproductive system in male offspring of rats given butyl benzyl phthalate during late pregnancy. AB - The objective of this study was to determine the effects of maternal exposure to butyl benzyl phthalate (BBP) on the development of the reproductive system in male offspring. Pregnant rats were given BBP by gastric intubation at 250, 500, or 1000 mg/kg on days 15 to 17 of pregnancy. A significant decrease in maternal body weight gain and food consumption was found in rats given BBP at 500 and 1000 mg/kg. A significant decrease in the number of live fetuses per litter was found at 1000 mg/kg. The weights of male and female fetuses were significantly decreased in the groups given BBP at 1000 mg/kg. A significant increase in the incidence of fetuses with undescended testes was found at 500 and 1000 mg/kg. A significant decrease in the anogenital distance (AGD) of male fetuses was observed at 500 and 1000 mg/kg. The AGD/cube root of body weight ratio in male fetuses was also significantly reduced at 500 mg/kg and higher. The AGD and AGD/cube root of body weight ratio of female fetuses in the BBP-treated groups were comparable to those in the control group. It was concluded that BBP on days 15 to 17 of pregnancy produced adverse effects on the development of the reproductive system in male offspring. PMID- 11934538 TI - Hepatic glutamine metabolism during endotoxemia in neonatal rats. AB - OBJECTIVES: The liver plays a central role during endotoxemia. We investigated the biochemical changes that occur in neonatal liver during early stages of endotoxemia. METHODS: Twenty neonatal rats (10 to 15 d; n = 10/group) were studied. Endotoxemic rats received intraperitoneal injections of 300 microg/kg of 12.5 mg/L of lipopolysaccharide and control rats received isovolemic normal saline. Two hours after injection, all lipopolysaccharide-injected animals exhibited signs of endotoxemia. Livers were removed and extracted into 12% perchloric acid. 1H and 31P magnetic resonance spectroscopy measured hepatic levels of glutamine, glutamate, alanine, lactate, glucose, beta-hydroxybutyrate, adenosine triphosphate, and adenosine diphosphate. Unpaired t test compared groups. RESULTS: No mortality occurred during the first 2 h after injection. Endotoxemia significantly decreased hepatic levels of glutamine (P < 0.001), glucose (P = 0.047), and beta-hydroxybutyrate (P < 0.001). There was no difference in hepatic levels of glutamate (P = 0.050), alanine (P = 0.165), lactate (P = 0.478), adenosine triphosphate (P = 0.165), and adenosine diphosphate (P = 0.136) between groups. CONCLUSIONS: Early endotoxemia caused significant changes in the hepatic metabolism of glutamine, glucose, and beta hydroxybutyrate. These findings increase our understanding of the pathophysiology of neonatal endotoxemia. PMID- 11934535 TI - Pregnancy outcome of women who failed appointments at a teratogen information service clinic. AB - The effect of failed appointments at a teratogen information service (TIS) clinic on pregnancy outcome is not known. The objectives of our study were 1) to identify predictors of failed appointments and 2) to evaluate if failed appointments were associated with a higher rate of adverse pregnancy outcome among women booked for counseling at a TIS clinic over 5 consecutive months. Of 242 patients scheduled to come to clinic, 154 were followed-up at 20 weeks gestational age. Women who were pregnant at 20 weeks were more likely to have attended the clinic (81% attendance rate), than women who had a spontaneous abortion, a therapeutic abortion, or an ectopic pregnancy (54% attendance rate) (P = 0.003). Independent predictors of failed appointments included exposure to illicit drugs, history of previous spontaneous abortion, and a main concern that was a nonmedication exposure. In conclusion, pregnant women who failed appointments at a TIS clinic are at greater risk for not being pregnant at 20 weeks gestational age than those who attended the clinic. Telephone counseling should be considered in selected cases to ensure that all women get relevant information at the time of appointment booking. PMID- 11934539 TI - Fatty acid oxidation in neonatal hepatocytes: effects of sepsis and glutamine. AB - OBJECTIVE: Little is known about fat use during sepsis during the neonatal period. Intramitochondrial O(2) consumption is inhibited in isolated hepatocytes from suckling septic rats and this impairment is reversed by glutamine. We investigated the effect of neonatal sepsis on fat oxidation and whether glutamine can directly affect fatty acid oxidation. METHODS: Suckling Wistar rats (11 d) received an intraperitoneal injection of 300 microg/kg of lipopolysaccharide (Escherichia coli 055:B5); controls received normal saline. At 2 h, hepatocytes were isolated. Hepatocytes were incubated at 37 degrees C with 0.5 mM [1 (14)C]palmitate or 0.5 mM [1-(14)C]palmitate plus 10 mM glutamine. After 1 h, the perchloric acid-soluble (14)C-radioactivity (representing mainly ketone bodies) and (14)CO(2) were measured. Hepatocyte O(2) consumption from 0.5 mM palmitate was measured with and without 2.5 ng/mL myxothiazol to estimate intramitochondrial O(2) consumption. RESULTS: There were no significant differences in fatty acid oxidation between control and endotoxemic hepatocytes measured as acid-soluble radioactivity (which represents mainly ketogenesis, plus Krebs cycle intermediates), as (14)CO(2) production, or as the sum of acid soluble radioactivity plus (14)CO(2) generation. Glutamine significantly increased fatty acid oxidation (acid-soluble radioactivity plus (14)CO(2)) in hepatocytes from control and endotoxic animals. CONCLUSIONS: The finding of no significant difference in fatty acid oxidation between hepatocytes from control and endotoxemic rats is surprising given that intramitochondrial O(2) consumption from palmitate is decreased. This may reflect altered use of acetyl-coenzyme A to ketone bodies and Krebs cycle intermediates. Glutamine enhanced fatty acid oxidation from control and endotoxemic hepatocytes, suggesting that it may promote substrate oxidation during endotoxemia. PMID- 11934540 TI - Hepatic glutamine metabolism. AB - Expression of high activities of both glutamine synthetase and glutaminase allows the liver to play a major role in the regulation of glutamine homeostasis. The liver shows net glutamine output in metabolic acidosis, in prolonged starvation and animals bearing tumors, net glutamine uptake in the postabsorptive state, on consuming high protein diets, and in uncontrolled diabetes or sepsis. Liver glutamine synthetase is expressed only in a small population of perivenous cells that allows it to salvage any ammonia not incorporated into urea in periportal cells. Hepatic glutaminase is a unique isozyme found only in periportal liver parenchymal cells where it provides glutamate and ammonia for the urea cycle. Control of hepatic glutamine metabolism occurs almost exclusively through changes in the activity of glutaminase, with no change in glutamine synthetase flux. PMID- 11934541 TI - Habitual food intake and polyunsaturated fatty acid deficiency in liver cirrhosis. AB - OBJECTIVE: We compared the habitual food intake and plasma fatty acid composition in cirrhotic patients living in two different regions in Japan, Okayama and Toyama, and evaluated the effects of dietary polyunsaturated fatty acid and alpha tocopherol intake on serum alanine aminotransferase (ALT) activity. METHOD: A quantitative food-frequency questionnaire method was used. RESULTS: The significantly higher intake of fish in the patients living in Toyama resulted in higher plasma levels of docosahexaenoic acid and lower levels of arachidonic acid. Serum ALT activity correlated negatively with plasma arachidonic acid (r = 0.456, P < 0.05) and alpha-tocopherol (r = -0.505, P < 0.05) levels. Dietary intakes of vitamin E and polyunsaturated fatty acids (mg/g) correlated negatively with serum ALT (r = -0.377, P < 0.05). Dietary intake of linoleic acid and the ratio of polyunsaturated to saturated fatty acid in dietary fat correlated significantly with serum ALT (r = 0.604, P < 0.01, and r = 0.622, P < 0.01, respectively). The amount of vegetable intake correlated with intake of vitamin E and polyunsaturated fatty acid (r = 0.527, P < 0.02). CONCLUSIONS: These findings suggest that habitual food intake affects the plasma fatty acid profile and that elevated serum ALT may be related to arachidonic acid deficiency and vulnerability to lipid peroxidation in cirrhotic patients with hepatitis B and C viruses. PMID- 11934542 TI - Circulating concentrations of soluble leptin receptor: influence of menstrual cycle and diet therapy. AB - Concentrations of the soluble leptin receptor (sOB-R) may be related to leptin resistance in obesity. We measured sOB-R concentrations in serum in 103 non diabetic Japanese men and women. All subjects were grouped according to body mass index (BMI; in kg/m(2)). Serum sOB-R concentrations did not differ significantly between normal-weight (18.5 < or = BMI < 25.0) men and women, but were significantly higher in underweight subjects (BMI < 18.5) than in normal-weight subjects. In contrast, overweight (25 < or = BMI < 30) and obese (30 < or = BMI < 35.0, 35.0 < or = BMI < 40, and BMI > or = 40) subjects had significantly lower sOB-R concentrations than did normal-weight subjects. Serum sOB-R concentrations were inversely correlated with BMI and serum immunoreactive leptin concentrations. Very low-energy diet therapy for 4 wk significantly lowered serum immunoreactive leptin concentrations but did not significantly affect serum sOB-R concentrations. Serum sOB-R concentrations did not change significantly during the menstrual cycle. Our results showed that serum sOB-R concentrations decrease with increasing BMI and that sex hormones likely do not affect serum sOB-R concentrations in non-pregnant women. The reduction in serum sOB-R concentrations in overweight and obese persons may reflect downregulation of hypothalamic leptin receptor production as a result of an increase in circulating leptin and might be an important factor in leptin resistance. PMID- 11934543 TI - Teaching medical students the subjective global assessment. AB - OBJECTIVE: Clinical nutrition assessment is a clinical skill not taught in many medical schools in North America. The purpose of this study is to determine whether second-year medical students can be taught to perform a nutritional Subjective Global Assessment (SGA). METHODS: In this study, second-year medical students were given a didactic session and a bedside demonstration of the SGA. Subsequently, they performed an SGA on unknown patients and classified those patients into one of three categories: A) well nourished, B) moderately malnourished, or C) severely malnourished. This was compared with the assessments of clinical dietitians and a physician. RESULTS: After this instruction, medical students correctly identified malnourished individuals. They were less accurate in their subclassification between mildly and severely malnourished individuals. The degree of agreement with clinical dietitians and a physician was fair (kappa = 0.34). CONCLUSIONS: With a multidisciplinary team of physicians and clinical dietitians, medical students can be taught the SGA in a 3h format. This is an important clinical skill that emphasizes the importance of clinical nutrition and may help identify malnourished individuals early in the course of their hospitalization. PMID- 11934544 TI - Transfer of selected elements from food into human milk. AB - OBJECTIVES: We measured the transfer factors of antimony, cerium, chromium, cobalt, gallium, lanthanum, molybdenum, niobium, ruthenium, silver, thorium, titanium, and uranium from food to milk in nursing mothers. METHODS: Food and milk samples from 19 mothers were taken daily over 2 to 8 wk. The samples were analyzed for element content after microwave-assisted pressure digestion with a mass spectrometer and inductively coupled plasma. The transfer factor, or the portion of element intake passed on in the milk, was calculated as the element concentration in food (g/kg) divided by the element concentration in milk (g/L). RESULTS: The calculated transfer factors were 5.1 for silver, 16.1 for cerium, 6.9 for chromium, 8.4 for cobalt, 19.1 for gallium, 13.8 for lanthanum, 77.4 for molybdenum, 20.7 for niobium, 4.1 for ruthenium, 13.2 for antimony, 20.2 for thorium, 5.6 for titanium, and 21.3 for uranium. Factors differed significantly across individuals. CONCLUSION: These differences can be attributed to the fact that the different levels of elements in breast milk are the result of individual differences in milk production and factors other than the amount of any particular element absorbed by the body. PMID- 11934545 TI - Use of positron emission tomography for the assessment of skeletal muscle glucose metabolism. AB - Positron emission tomography (PET) is a unique tool for studying regional skeletal muscle glucose metabolism and blood flow in vivo. The application of PET in the assessment of skeletal muscle glucose metabolism depends on recent improvements in instrumentation, data analysis, and production of (18)F fluorodeoxyglucose (FDG) and (15)O water. The data presented support the validity of the (15)O water model to measure blood flow and the FDG model for the determination of glucose uptake and glucose kinetic constants (influx, efflux, and phosphorylation) in skeletal muscle. However, quantification of absolute glucose transport and backflux rates should be applied with caution because those calculations are based on unproven assumptions such as validity of the lumped constant for these individual processes and constancy of the free and accessible intracellular glucose pool. It is evident that quantification of glucose fluxes using the triple tracer technology generates conflicting data that violate assumptions inherent in triple tracer or PET modeling. Further FDG-PET studies will have to solve those problems to provide more insight into the regulatory processes of glucose transport and phosphorylation of different insulin-resistant disease states. Promising new areas of PET research will include not only detailed study of glucose kinetics but also the measurement of muscle protein synthesis in vivo, which is of interest in a variety of conditions. PMID- 11934546 TI - Glutamine and arginine affect Caco-2 cell proliferation by promotion of nucleotide synthesis. AB - OBJECTIVE: We tested our hypothesis that 1) the major effect of Gln is as a nitrogen donor, not an energy source, for nucleotides (NT) and 2) the supplementation of culture medium with arginine (Arg) decreases the flux of glutamine (Gln) for conversion to Arg, thus accelerating NT synthesis. METHODS: Various concentrations of nucleosides (NS+NT) Gln, and glutamate (Glu) in culture were tested for their effect on Caco-2 cell proliferation. (Arg was tested in media with and without Gln to evaluate the Gln pathway. The incorporation of (15)N from L-[5-(15)N]-Gln into NTs of DNA was measured under different NS + NT and Arg concentrations.) RESULTS: The proliferation of Caco-2 cells was increased by NS + NT and Gln supplementation, but not by Glu. The effective concentration of NS + NT was 100-fold smaller than that of Gln. An Arg effect was observed only in the presence of Gln. The NT synthesis from Gln, as indicated by (15)N incorporation from L-[5-(15)N]-Gln, was increased by Arg supplementation and decreased by NS + NT supplementation. CONCLUSION: These results support our hypothesis that the effects of Gln and Arg on Caco-2 cell proliferation are by the promotion of NT synthesis and that the major role of Gln is not energy supply. PMID- 11934547 TI - Oral glutamine does not prevent bacterial translocation in rats subjected to intestinal obstruction and Escherichia coli challenge but reduces systemic bacteria spread. AB - OBJECTIVE: We investigated whether oral glutamine prevents bacterial translocation. METHODS: Male Wistar rats were fed with isocaloric and isoproteic standard rat chow and randomly assigned to receive glutamine (GLN) or glycine administered through an orogastric tube at 1.5 g.kg(-1).d(-1) for 7 d. On day 8 of the study, the animals were anesthetized and intestinal obstruction was produced by ligature of the terminal ileum. A suspension containing 10(9) colony forming units per milliliter of Escherichia coli ATCC 25992 was injected into the lumen of the ileum. Twenty-four hours later, blood was withdrawn, and mesenteric lymph nodes and fragments of spleen, liver, and lung were sent for microbiological analysis. Cultures were done on blood agar and MacConkey agar. Student's t test and analysis of variance between two proportions were used. P < 0.05 was considered significant. RESULTS: Rats in both groups lost body weight during the experiment (not significant). Mesenteric lymph node cultures were positive in both groups. The GLN group had a smaller percentage of E. coli in blood and organ cultures (65.45% versus 82.67% in the glycine group; P = 0.027). Positive cultures of blood, spleen, liver and lung also were higher on glycine group, although not significantly. CONCLUSIONS: Oral GLN does not prevent bacterial translocation in rats after intestinal obstruction and E. coli challenge. No specific organ was protected by GLN. Nevertheless, its use was associated with a reduced number of positive E. coli cultures in blood and remote organs, and thus diminished bacteria spread. This association suggests a role for GLN in gut barrier protection, possibly by immune system enhancement. PMID- 11934548 TI - Roles of nucleosides and nucleotide mixture in small bowel transplantation. AB - OBJECTIVES: We investigated the effect of nucleosides (NSs) and nucleotides (NTs) on the intestine and intestinal graft in a model of syngenic small bowel transplantation, with the fetal rat intestine as a graft. METHODS: Two-centimeter jejunal segments from Lewis rats at 19 d of gestation were transplanted into the abdominal walls of 5-wk-old Lewis rats by using a non-vascular anastomotic technique. After transplantation, the rats were assigned to one of two groups: group 1 did not receive NS or NT and group 2 was supplemented with NS and NT. The grafts and graft recipients were examined morphologically 14 d after transplantation according to conventional histologic and immunohistochemical studies of neurons and smooth muscles. RESULTS: Group 1 gained little body weight, even though both groups received similar amounts of food. The grafts in group 1 showed poor development in length, diameter, and wet weight. They also showed poor villi development, abnormalities in nerve distribution, and degeneration of muscle layer structure on histologic and immunohistochemical studies. CONCLUSIONS: We found that NS and NT are essential nutrients for intestinal growth and maintenance of structures in fetal small bowel transplantation. PMID- 11934549 TI - Nutrition and disease: challenges of research design. AB - This review discusses research methodology in the relation between diet and disease. Medical research can be divided into two types: complex research (the detailed study of disease mechanisms using such methods as biochemistry and molecular genetics) and simple research (the investigation of the factors that cause or prevent disease using methods such as epidemiology, intervention trials, and analagous studies on animals). Although complex research has received the bulk of resources, the large majority of our information of practical value has come from simple research. This general principle is illustrated in the area of diet and disease by examples from various areas: selenium, carotenoids, and cancer; vitamin E, omega-3 fatty acids, and coronary heart disease; dietary fat and obesity; dietary sodium and hypertension; and alcohol and stroke. Discussion then turns to aspects of the design of cohort (prospective) studies. Because of problems of sample size and relative lack of diversity, previous studies often failed to give clear-cut results. Suggestions are made concerning the design of cohort studies, notably the use of much larger numbers of subjects and with greater diversity in their diets. The problem of confounding also is discussed. Lifestyle factors often cluster together but cohort studies may not have fully unraveled this. PMID- 11934550 TI - Effect of essential fatty acids and their metabolites on human lymphocytic leukemia and human colon adenocarcinoma lymph node cells in vitro. PMID- 11934551 TI - Fat emulsion and stress response in surgical patients. PMID- 11934552 TI - What types of nutrition research give the best benefits? PMID- 11934553 TI - Intestinal zinc, iron, and copper during the perinatal period. PMID- 11934554 TI - Zinc intake versus zinc absorption: a bioavailability factor. PMID- 11934556 TI - The nutrition situation in Uganda. PMID- 11934557 TI - Making friends with the fear. PMID- 11934558 TI - A trawl through the current nutrition literature. PMID- 11934559 TI - Supplementation and the elderly: dramatic results? PMID- 11934561 TI - Parenteral and mucosal prime-boost immunization strategies in mice with hepatitis B surface antigen and CpG DNA. AB - Synthetic oligodeoxynucleotides (ODN) containing immunostimulatory CpG motifs (CpG ODN) are potent adjuvants to protein antigens administered by parenteral or mucosal routes to BALB/c mice. To date, there have been no studies using combined parenteral/mucosal approaches with CpG DNA as adjuvant. In this study we evaluated different parenteral prime-mucosal boost and mucosal prime-parenteral boost strategies using hepatitis B surface antigen (HBsAg) alone or with different adjuvants: aluminum hydroxide (alum), cholera toxin (CT), CpG ODN. In addition, since CpG ODN has previously been shown to act synergistically with other adjuvants after parenteral or mucosal delivery, we also evaluated adjuvant combinations: alum+CpG ODN and CT+CpG ODN. The effects of adjuvant and administration strategy on systemic and mucosal humoral responses were measured, as well as cell-mediated immune responses (cytotoxic T lymphocyte activity). These results were compared to parenteral only or mucosal only strategies. Our findings demonstrate that parenteral immunization can prime for mucosal responses even when different lymph nodes were being targeted. HBsAg-specific immune responses (IgG in plasma, cytotoxic T lymphocytes) induced by parenteral prime could all be significantly enhanced by mucosal boosting and despite the fact that intramuscular immunization alone could not induce mucosal IgA, it could prime for a subsequent mucosal boost. In addition, the presence of adjuvant at time of boosting could influence the nature of subsequent immune responses (Th1 vs. Th2). Mice primed intranasally could have their systemic immune responses boosted with a parenteral administration and it was also possible to enhance mucosal responses induced by intranasal prime with an intramuscular boost. PMID- 11934562 TI - N-acetyl-D-glucosamine specific hemagglutinin receptor of Vibrio cholerae O1 in chicken erythrocyte membranes. AB - N-Acetyl-D-glucosamine specific cell-associated hemagglutinin (HA)/lectin, previously purified from a strain of Vibrio cholerae O1, had been established as an adhesin molecule of V. cholerae O1 cells. This communication records the isolation and purification of the glycoprotein receptor of the N-acetyl-D glucosamine specific HA of the V. cholerae O1 strain from chicken erythrocyte membranes. The most salient feature of this study is that the pretreatment of partially purified glycoprotein with purified HA could completely inhibit the hemagglutinating activity of the V. cholerae O1 strain with chicken erythrocytes. PMID- 11934563 TI - Characteristics of Staphylococcus aureus, isolated from airways of cystic fibrosis patients, and their small colony variants. AB - The colonization of respiratory tract by Staphylococcus aureus is a frequent feature of cystic fibrosis (CF), especially in pediatric patients. The formation of small colony variants (SCVs), which produce reduced amounts of alpha-toxin, is one of the proposed ways of staphylococcal accommodation in an intracellular niche. The aim of the present study was to compare some properties of S. aureus SCVs and their parent strains. A site-directed S. aureus hemB mutant and parent strain 8325-4 were included in the study (control pair). Normal and SCV strain pairs from CF patients as well as control strains were tested for the susceptibility to defensins, killing activity of professional phagocytes and adhesion to A549 cell line. Because S. aureus are exposed to many cationic proteins in the host, we challenged a clinical isolate with minimal subinhibitory concentration (subMIC) of protamine and found that hemin and menadione auxotrophic SCVs emerged. SCVs were more resistant than normal strains to protamine but not to dermaseptin. The susceptibility to the bactericidal activity of magainin was the same for normal and SCV strains. The protamine resistance of normal as well as SCVs was strongly enhanced by high salt concentration. The adhesion of some SCVs to A549 cells was higher than adhesion of parental strains. However, the number of adherent bacteria (SCVs) was diminished in the presence of hemin for hemin auxotrophs. The uptake of SCVs by granulocytes was lower than ingestion of normal strains, but SCVs were killed with equal or greater potency. SCVs are adapted to intracellular survival and persistence in the host under certain circumstances. The ability to form a variant subpopulation affords S. aureus additional survival options. PMID- 11934564 TI - Changes in cell-mediated immunity in kidney transplant recipients with active CMV infection. AB - This study was aimed at determining (a) the extent of proliferation of peripheral blood mononuclear cells (PBMC) in response to stimulation by cytomegalovirus (CMV)-infected fibroblasts and (b) the levels of Th1 and Th2 cytokine production in kidney transplant recipients with and without active CMV infection. Thirty patients with, and 39 without active CMV infection, diagnosed by a CMV antigenemia assay (AA), were studied. PBMC of patients with active CMV infection showed significantly lower proliferation than those without ongoing CMV infection (P<0.0001). The levels of Th2-type cytokines (interleukin (IL-) 4 and IL-10) in AA-negative and AA-positive kidney transplant recipients were similar but the levels of the Th1-type cytokines interferon-gamma, tumor necrosis factor-alpha (P<0.05) and IL-2 were significantly lower in AA-positive kidney transplant recipients (P<0.0005). PMID- 11934565 TI - The antibody response to bacteriophage is linked to the lymphopenia gene in congenic BioBreeding rats. AB - Congenic BioBreeding (BB) rats, homozygous for the autosomal lymphopenia (Lyp) gene (Lyp/Lyp), heterozygous (Lyp/+), or wild-type (+/+), were immunized with the T cell-dependent bacteriophage PhiX174 to determine effects of Lyp on primary and secondary antibody responses. The primary PhiX174 antibody response did not differ between the three different genotypes. In contrast, the secondary immune response, expressed as the peak neutralizing titer, was markedly reduced in Lyp/Lyp (9.9+/-3.2; mean value+/-S.E.M. for seven rats) compared to both Lyp/+ (51+/-12; n=13; P=0.006) and +/+ (100+/-20; n=7; P=0.004) BB rats. We suggest that the secondary antibody response to the T cell-dependent neoantigen PhiX174 is linked in a recessive manner to genetic factor(s) in the Lyp gene region. PMID- 11934566 TI - Binding of Clostridium difficile to Caco-2 epithelial cell line and to extracellular matrix proteins. AB - Adhesion of Clostridium difficile to Caco-2 was examined as a function of monolayers polarization and differentiation. The number of adherent C. difficile C253 bacteria per cell strongly decreased when postconfluent 15-day-old monolayers were used (1.7 bacteria per cell versus 17.3 with 3-day-old monolayers). Following disruption of intercellular junctions by ethylene glycol bis(beta-aminoethyl ether)-N,N,N',N',-tetraacetic acid, a significant rise in the level of bacterial adhesion was observed, above all in postconfluent monolayers. Immunofluorescence studies of bacteria and transferrin receptor, a marker of basolateral pole of polarized monolayers, showed that C. difficile C253 adheres mainly to the basolateral surface of differentiated and undifferentiated polarized Caco-2 cells. Furthermore, binding of C. difficile C253 to several extracellular matrix proteins in vitro was demonstrated by an ELISA-based assay. PMID- 11934567 TI - The expression of the Helicobacter pylori genes ureA and nap is higher in vivo than in vitro as measured by quantitative competitive reverse transcriptase-PCR. AB - The expression of the virulence-associated genes ureA, encoding the urease subunit A, and nap, encoding the neutrophil activating protein, in Helicobacter pylori grown both in the stomach of C57/Bl6 mice and in Brucella broth was quantified by quantitative competitive reverse transcriptase-PCR using a homologous RNA standard (competitor) and an external standard (16S rRNA). The results showed that the ureA and nap transcripts were increased up to 15 and 80 times, respectively, in vivo compared to in vitro. The transcription of ureA and nap also differed in that ureA showed highest expression early in infection in mice whereas nap transcription was variable throughout the 18-week infection period. PMID- 11934568 TI - Variations among clinical isolates of Staphylococcus aureus to induce expression of E-selectin and ICAM-1 in human endothelial cells. AB - Eighteen clinical isolates of Staphylococcus aureus, nine methicillin-sensitive and nine methicillin-resistant, were investigated for their ability to induce expression of E-selectin and ICAM-1 in human endothelial cells. Upregulation of adhesion molecules varied between isolates; 17 isolates induced expression of E selectin and 13 of ICAM-1. Some isolates induced a significant expression of E selectin without stimulation of ICAM-1, whereas the opposite was not found. Bacterial viability was required for induction of the adhesion molecules. The kinetics of ICAM-1 expression in S. aureus-infected cells differed from those stimulated with interleukin-1beta (IL-1beta). On the other hand, expression of E selectin was very similar in S. aureus-infected and IL-1beta-stimulated cells. There was no correlation between ability of S. aureus to induce expression of cell adhesion molecules, methicillin susceptibility, pulse field gel electrophoresis patterns, biochemical characteristics, phage typing and toxin production. PMID- 11934569 TI - Incidence of toxic Aeromonas isolated from food and human infection. AB - One hundred and ninety four Aeromonas isolates (99 from food and 95 from clinical sources) were analyzed as to the species involved and the toxins produced. Of the clinical isolates of Aeromonas, 29.4% were enterotoxigenic, 43.1% were hemolytic and 89% were cytotoxigenic. Among the food isolates, 18.2% were enterotoxigenic, 17.1% were hemolytic and 72.7% were cytotoxigenic. Aeromonas sobria and Aeromonas veronii produced more enterotoxin and cytotoxin than the other isolates, whereas A. veronii and Aeromonas salmonicida produced cell-free hemolysin. Most of the isolates produced cytotoxins (81%) active on Vero (green monkey kidney) and Chinese hamster ovary cells, but only the culture supernatant of A. sobria produced vacuolation in these cell lines. PMID- 11934570 TI - Potential virulence and antimicrobial susceptibility of Aeromonas popoffii recovered from freshwater and seawater. AB - Aeromonas popoffii is the most recent species within the genus Aeromonas described from freshwater. In our study this species was also recovered from this habitat and for the first time from seawater. Most of the virulence factors known in Aeromonas spp. (aerolysin/hemolysin, serine protease, lipases and DNases) were highly prevalent in this species. Third-generation cephalosporins and quinolones were the most active antimicrobial agents against A. popoffii. PMID- 11934571 TI - Characterisation of 20-kDa lectin-spermagglutinin from Arum maculatum that prevents Chlamydia pneumoniae infection of L-929 fibroblast cells. AB - A novel lectin from the root of Arum maculatum was isolated by saline extraction and purified by cold ethanol precipitation and subsequent fractionation on Superose 6 column. The lectin named A. maculatum agglutinin is a non-glycosylated protein with 20-kDa molecular mass agglutinating human ejaculated spermatozoa, but not human erythrocytes. The agglutination was blocked in the presence of N acetylneuraminic acid indicating that the lectin is sialoglycoprotein specific. Chlamydia pneumoniae strain AR-39 showed considerable potential to grow in murine L-929 fibroblast cells. Pretreatment of the cell monolayers with purified lectin reduced the entry and intracellular replication of C. pneumoniae. These results suggest that the isolated lectin prevents attachment by binding to a C. pneumoniae specific sialoglycoprotein receptor expressed on the surface of L-929 fibroblast cells. PMID- 11934573 TI - An efficient chemical synthesis of nicotinamide riboside (NAR) and analogues. AB - A simple and efficient synthesis of nicotinamide riboside (NAR) 1 and derivatives 4 and 5 via trimethylsilyl trifluoromethanesulfonate (TMSOTf)-mediated N glycosilation followed by spontaneous deacetylation by treating with methanol is reported. PMID- 11934574 TI - Design and synthesis of lignostilbene-alpha,beta-dioxygenase inhibitors. AB - Lignostilbene-alpha,beta-dioxygenase cleaves the olefinic double bond of phenolic stilbenes by a mechanism similar to that of 9-cis-epoxycarotenoid dioxygenase, a key enzyme in abscisic acid biosynthesis. Several analogues of stilbene were designed and synthesized, and their efficacy as inhibitors of lignostilbene alpha,beta-dioxygenase was examined. The compound (Z)-1-(4-hydroxyphenyl)-1 fluoro-2-phenylethene (2) was found to be a potent inhibitor of this enzyme with an IC(50) of 3 microM. PMID- 11934575 TI - A practical approach to the synthesis of hairpin polyamide-peptide conjugates through the use of a safety-catch linker. AB - Hairpin polyamides are high-affinity, sequence selective DNA binders. The use of a safety-catch linker for the solid phase synthesis of hairpin polyamides allows for easy preparation of derivatives ready for chemoselective ligation with unprotected peptides. Examples of ligations reported include thioether bond formation and thioester-mediated amide bond formation ('Native Chemical Ligation'). PMID- 11934576 TI - Synthesis and pharmacological evaluation of 6-piperidino- and 6-piperazinoalkyl 2(3H)-benzothiazolones as mixed sigma/5-HT(1A) ligands. AB - In an effort to produce new pharmacological probes with mixed sigma/5-HT(1A) affinity, we have synthesized a series of 12 original 6-piperidino- or piperazino alkyl-2(3H)-benzothiazolones and their receptor binding profile (sigma, 5-HT(1A), 5-HT(2A), 5-HT(3), D(2), H(1), and M(1)) was determined. The best mixed sigma/5 HT(1A) affinity profile was found within the piperidine series with 4-benzyl substitution associated to linker methylene chain n=2 (K(i) 5 and 4nM, respectively). Moreover, a highly selective sigma2 ligand was obtained with a 3,4 dichlorobenzyl substitution associated to n=4 (K(i) 2nM, selectivity ratio sigma1/sigma2=70). PMID- 11934577 TI - New synthetic analogues of N-acyl homoserine lactones as agonists or antagonists of transcriptional regulators involved in bacterial quorum sensing. AB - A series of 22 novel synthetic N-acyl-homoserine lactone analogues has been evaluated for both their inducing activity and their ability to competitively inhibit the action of 3-oxo-hexanoyl-L-homoserine lactone, the natural inducer of bioluminescence in the bacterium Vibrio fischeri. In the newly synthesized analogues, the extremity of the acyl chain was modified by introducing ramified alkyl, cycloalkyl or aryl substituents at the C-4 position. Most of the analogues bearing either acyclic or cyclic alkyl substituents showed inducing activity. In contrast, the phenyl substituted analogues displayed significant antagonist activity. We hypothesized that the antagonist activity of the phenyl compounds may result from the interaction between the aryl group and aromatic amino acids of the LuxR receptor, preventing it from adopting the active dimeric form. PMID- 11934578 TI - Solid-phase synthesis and pharmacological evaluation of analogues of PhTX-12-A potent and selective nicotinic acetylcholine receptor antagonist. AB - Philanthotoxin-12 (PhTX-12) is a novel potent and selective, noncompetitive antagonist of nicotinic acetylcholine receptors (nAChRs). Homologues of PhTX-12 with 7-11 methylene groups between the primary amino group and the aromatic head group were synthesized using solid-phase methodology. In vitro electrophysiological studies of nAChR demonstrated that decreasing the number of methylene groups from 12 to 11 significantly increased potency. Antagonism by PhTX-11, like that of PhTX-12, was only weakly voltage-dependent. When the methylene spacer was reduced further, antagonism was decreased below that of PhTX 12, and in some cases potentiation of ACh responses by up to 60% was observed. PMID- 11934579 TI - Neurotrophic activity of honokiol on the cultures of fetal rat cortical neurons. AB - Honokiol, a main biphenyl neolignan of the traditional crude medicine, Magnoliae cortex, was found to show neurotrophic activity on the cultures of rat cortical neurons at concentration from 0.1 to 10 microM. In the cortical neurons cultured in serum-free medium supplemented with B27, honokiol could promote neurite outgrowth. In addition, the survival and growth of neurons were significantly enhanced by adding honokiol to the primary cultures in serum-free medium supplemented with N2. Its neurotrophic activity was comparable to 40 ng mL(-1) of bFGF at concentration of 10 microM. PMID- 11934580 TI - Enzymatic synthesis of labeled DNA by PCR using new fluorescent thymidine nucleotide analogue and superthermophilic KOD dash DNA polymerase. AB - Triphosphate of a new fluorescent labeled thymidine analogue was incorporated as a substrate for PCR using KOD Dash DNA polymerase forming the corresponding fluorescent labeled DNA which is useful for a DNA probe. PMID- 11934581 TI - Novel potent antagonists of human neuropeptide Y Y5 receptor. Part 1: 2 oxobenzothiazolin-3-acetic acid derivatives. AB - Novel NPY-Y5 antagonist FR73966 was discovered by screening of our in-house chemical library. The analogues were prepared by application of parallel synthesis techniques. Some of the resulting 2-oxobenzothiazolin-3-acetic acid derivatives exhibited nanomolar binding affinity for human NPY-Y5 receptors. PMID- 11934582 TI - Carbonic anhydrase activators: human isozyme II is strongly activated by oligopeptides incorporating the carboxyterminal sequence of the bicarbonate anion exchanger AE1. AB - Di-/tri- and especially tetrapeptides incorporating the sequence DADD present in the carboxyterminal region of the bicarbonate/chloride anion exchanger AE1 strongly activate human carbonic anhydrase (CA) isozyme II, whereas they act as more inefficient activators of isozymes I and IV. This discovery suggests that in the metabolon hCA II-AE1, the last protein plays a role both as a CA activator as well as a bicarbonate transporter. A synthesis of the tripeptide DAD and the tetrapeptide DADD is also presented together with the possible explanation why such highly acidic oligopeptides efficiently bind to hCA II but not to the closely related isozymes I and IV. PMID- 11934583 TI - Novel bicyclic lactam inhibitors of thrombin: highly potent and selective inhibitors. AB - The potency and selectivity of a previous series of low molecular weight thrombin inhibitors were improved through modifications of the P1 and P3 residues. Introduction of diphenyl substituted sulfonamides in the P3 moiety led to highly efficacious compounds. By correctly selecting the combination of P1 and P3 residues, high levels of potency, selectivity and in vivo efficacy were obtained. PMID- 11934584 TI - Synthesis and structure-activity relationships of a novel series of HIV-1 protease inhibitors encompassing ABT-378 (Lopinavir). AB - The HIV protease inhibitor ABT-378 (Lopinavir) has a 2,6-dimethylphenoxyacetyl group in the P-2' position. Analogues in which this group is replaced with various substituted phenyl or heteroaryl groups were synthesized and the structure-activity relationships explored. PMID- 11934585 TI - Synthesis and in vitro evaluation of a novel iodinated resiniferatoxin derivative that is an agonist at the human vanilloid VR1 receptor. AB - Using a 'directed' iodination procedure, novel iodo-resiniferatoxin congeners were synthesized from 4-acetoxy-3-methoxyphenylacetic acid and resiniferinol- 9,13,14-ortho-phenylacetate (ROPA). The 2-iodo-4-hydroxy-5-methoxyphenylacetic acid ester of resiniferinol 5 displayed high affinity binding (K(i)=0.71 nM) for the human vanilloid VR1 receptor and functioned as a partial agonist. PMID- 11934586 TI - Anisylazoformylarginine: a superior assay substrate for carboxypeptidase B type enzymes. AB - Anisylazoformylarginine (CH(3)OC(6)H(4)-N=N-CO-Arg-OH) is rapidly hydrolyzed at the acyl-arginine linkage by the zinc-enzyme porcine carboxypeptidase B. The catalytic reaction is readily monitored spectrophotometrically by disappearance of the intense absorption (349 nm, epsilon 19,100) of the azo chromophore, which chemically fragments after substrate cleavage. PMID- 11934587 TI - Anthranilate sulfonamide hydroxamate TACE inhibitors. Part 1: Structure-based design of novel acetylenic P1' groups. AB - The structure-based design of potent sulfonamide hydroxamate TACE inhibitors bearing novel acetylenic P1' groups has led to compounds with excellent in vitro potency against TACE and selectivity over MMP-1. PMID- 11934588 TI - Anthranilate sulfonamide hydroxamate TACE inhibitors. Part 2: SAR of the acetylenic P1' group. AB - The SAR of a series of potent sulfonamide hydroxamate TACE inhibitors bearing novel acetylenic P1' groups was explored. In particular, compound 4t bearing a butynyloxy P1' moiety has excellent in vitro potency against isolated TACE enzyme and in cells, good selectivity over MMP-1 and oral activity in an in vivo model of TNF-alpha production. PMID- 11934589 TI - Non-covalent thrombin inhibitors featuring P(3)-heterocycles with P(1)-monocyclic arginine surrogates. AB - Investigations on P(2)-P(3)-heterocyclic dipeptide surrogates directed towards identification of an orally bioavailable thrombin inhibitor led us to pursue novel classes of achiral, non-covalent P(1)-arginine derivatives. The design, synthesis, and biological activity of inhibitors NC1-NC30 that feature three classes of monocyclic P(1)-arginine surrogates will be disclosed: (1) (hetero)aromatic amidines, amines and hydroxyamidines, (2) 2-aminopyrazines, and (3) 2-aminopyrimidines and 2-aminotetrahydropyrimidines. PMID- 11934590 TI - Synthesis of sub-micromolar inhibitors of MraY by exploring the region originally occupied by the diazepanone ring in the liposidomycin structure. AB - The synthesis and inhibitory activity against MraY of a series of simplified analogues of liposidomycins are described. These compounds were mainly obtained by performing parallel synthesis in the 6'-position of a scaffold that gathers key features found necessary for the binding to MraY. Thus, inhibitory activity was improved from 5300 to 140 nM. This improvement was correlated with the length and lipophilicity of substituents, but was found to be independent of the nature of the chemical bond generated. In addition, some of these inhibitors presented encouraging antibacterial activities. PMID- 11934591 TI - Novel matrix metallo-proteinase (MMP-2) phosphonoboronate inhibitors. AB - A series of novel phosphonoboronates consisting of PC(1)B, PC(n)B, PC(X)C(n)B, and PCC=CB derivatives were evaluated as MMP-2 inhibitors. Structure-activity relationships (SARs) data for the compounds were discovered and are discussed. PMID- 11934592 TI - Photochemical preparation of a pyridone containing tetracycle: a Jak protein kinase inhibitor. AB - Jak3 is a protein tyrosine kinase that is associated with the shared gamma chain of receptors for cytokines IL2, IL4, IL7, IL9, and IL13. We have discovered that a pyridone-containing tetracycle (6) may be prepared from trisubstituted imidazole (5) in high yield by irradiation with >350 nm light. Compound 6 inhibits Jak3 with K(I)=5 nM; it also inhibits Jak family members Tyk2 and Jak2 with IC(50)=1 nM and murine Jak1with IC(50)=15 nM. Compound 6 was tested as an inhibitor of 21 other protein kinases; it inhibited these kinases with IC(50)s ranging from 130 nM to >10 microM. Compound 6 also blocks IL2 and IL4 dependent proliferation of CTLL cells and inhibits the phosphorylation of STAT5 (an in vivo substrate of the Jak family) as measured by Western blotting. PMID- 11934593 TI - A practical synthesis of (S) 3-tert-butoxycarbonylamino-2-oxo-2,3,4,5-tetrahydro 1,5-benzodiazepine-1-acetic acid methyl ester as a conformationally restricted dipeptido-mimetic for caspase-1 (ICE) inhibitors. AB - A simple and versatile method for the synthesis of (S) 3-tert-butoxycarbonylamino 2-oxo-2,3,4,5-tetrahydro-1,5-benzodiazepine-1-acetic acid methyl ester (4), a dipeptide mimetic, has been developed. The regioselective functionalization of the N1 and N5 ring nitrogens and the C3 amino group is demonstrated in the synthesis of an interleukin-1beta converting enzyme inhibitor 13. PMID- 11934594 TI - Self-assembly of synthetic zinc chlorins in a silicate micelle prepared by sol gel process. AB - Zinc methyl 3-devinyl-3-hydroxymethyl-pyropheophorbide-a (1), a good model compound of light-harvesting pigments of green photosynthetic bacteria, formed self-aggregates in the presence of octadecyltriethoxysilane and tetraethoxysilane in an aqueous solution to exhibit visible absorption spectra similar to the natural antenna. Base-catalyzed cross-linked polymerization of the additive Si ORs (R=ethyl and H) afforded the formation of a siloxane network (Si-O-Si) on the surface of the self-assemblies of 1. The resulting microcapsules were stable to tolerate the deaggregation to monomeric 1 by addition of surfactant Triton X-100 more largely than the corresponding micelles before polymerization. PMID- 11934595 TI - Amino acid derived sulfonamide hydroxamates as inhibitors of procollagen C proteinase. Part 2: Solid-phase optimization of side chains. AB - Optimization of the amino acid side chain and the N-alkyl group of the sulfonamide of amino acid derived sulfonamide hydroxamates is discussed. The solid-phase synthesis of these potent inhibitors of procollagen C-proteinase (PCP) is presented. In addition, novel carboxylic acid sulfonamides were discovered to be PCP inhibitors. PMID- 11934597 TI - Advances towards understanding heart valve response to injury. AB - BACKGROUND: Composed of endocardial endothelial, valvular interstitial, cardiac muscle, and smooth muscle cells (SMC), heart valves are prone to various pathologic conditions the morphology of which has been well described. The morphology of diseased valves suggest that the "response to injury" process occurs in these valves, and is associated with an accumulation of interstitial cells and matrix, valvular inflammation and calcification, conditions that lead to dysfunction. The purpose of this study is to describe the current knowledge of the regulation of the valvular "response to injury" process, since we feel that this paradigm is essential to understanding valve disease. METHODS: The pertinent literature relating to the cell and molecular biology of valvular repair, and specifically interstitial cell function in valve repair, is reviewed. RESULTS: The cell and molecular biology of valve interstitial cells are poorly understood. Molecules regulating some of the aspects of the "response to injury" process have been studied, however, the signal transduction pathways, gene activation, and interactions of bioactive molecules with each other, with cells, and with the matrix have not been characterized. Initial studies identify the cell and molecular biology of interstitial cells to be an important area of research. Agents that have been studied include nitric oxide (NO) and FGF-2 and several matrix-related proteins including osteopontin. The present review suggests several directions for future study and a working model of valvular repair is presented. DISCUSSION: The regulation of the "response to injury" process in the human heart valve is still largely unknown. The cell and molecular events and processes that occur in heart valve function and repair remain poorly understood. These events and processes are vital to our understanding of the pathobiology of heart valve disease, and to the successful design of tissue engineered replacement valves. PMID- 11934598 TI - Differential integrin expression by cardiac fibroblasts from hypertensive and exercise-trained rat hearts. AB - The cardiac fibroblast is the principal cell type responsible for extracellular matrix (ECM) synthesis in the heart during growth and pathophysiological conditions. A dynamic interaction exists between the cardiac ECM and fibroblasts that is sensitive to the local mechanical and chemical tissue environment. We propose here that cardiac fibroblasts structurally and functionally adapt to changing local environments by altering their expression of receptor integrins. Changes in the extracellular environment are communicated in part by integrins, which link the ECM to the cell and regulate phenotype and function. In this report, we analyze integrin protein expression, migration and gel contraction by cardiac fibroblasts from rats subjected to 10 weeks of treadmill exercise (XTR), experimental hypertension (HYP) or controls (CONT). Immunoprecipitation shows that beta1 protein increases in XTR and HYP. Also, alpha1 and alpha2 integrins are lower in XTR and HYP, and alpha5 integrin is higher in XTR and lower in HYP. Functional assays show that XTR and HYP migrate slower on collagen, while XTR migrate faster and HYP slower on fibronectin. Cell isolation procedure, population expansion number or a general adaptation to culture conditions does not explain the differences observed. No significant differences in collagen gel contraction are detected. These results indicate that cardiac fibroblasts retain their in vivo patterns in vitro for a limited number of population expansions. This tissue-specific phenotype is exhibited in early passage (< or =6). However, by late passage (>8), cells begin to show adaptation to the in vitro conditions. These results show that cardiac fibroblasts respond to changing environments in pathophysiological conditions by modulating integrin expression, which is associated with changes in cell migration. They also suggest a pragmatic use for primary cardiac fibroblasts as a model to study the cardiac matrix remodeled by physiological (exercise) and pathological (hypertension) stressors. PMID- 11934599 TI - Ventricular remodeling after myocardial infarction and effects of ACE inhibition on hemodynamics and scar formation in SHR. AB - The effect of ACE inhibition after myocardial infarction (MI) on MI healing and remodeling in the presence of hypertension is not exactly known. Therefore, the effect of quinapril on scar formation, remodeling and hemodynamics was studied in spontaneously hypertensive rats (SHR). Nine weeks after moderate and large MI, left ventricular end-diastolic pressure (LVEDP) and passive pressure-volume relations were similar in 28-week-old hypertensive and normotensive rats. Chronic therapy with quinapril (6 mg/kg/day, started 30 min post-MI) reduced LVEDP and LV to body weight ratio, yet did not affect pressure-volume relations. Quinapril increased MI size and reduced the content and brightness of collagen fibers in the scar examined by polarized light microscopy. In conclusion, ventricular dilatation after MI was not accelerated in SHR, probably due to LV hypertrophy. Quinapril produced beneficial hemodynamic effects similar to that observed in the normotensive rat model. The significance and timing of ACE inhibitor-induced impairment of scar formation need further evaluation. PMID- 11934600 TI - Left ventricular aneurysm, aortic valve disease and coronary narrowing in a patient with Hunter's syndrome. AB - Hunter's syndrome (mucopolysaccharidosis type 2, MPS 2) is an inherited disorder of glycosaminoglycan degradation commonly associated with cardiac disease. We present the case of a young man with unusual cardiac manifestations of this syndrome. When mixed aortic valve disease was noted in childhood, other classical features of the milder form of Hunter's syndrome were present. There was no symptomatic or echocardiographic cardiovascular deterioration until age 27 when the patient presented in severe biventricular failure. Investigations demonstrated cardiomegaly and a large apical left ventricular aneurysm. The patient died suddenly soon after this diagnosis. Post mortem examination demonstrated a hypertrophied left ventricle with a 6-cm apical aneurysm. Coronary arterial walls were diffusely thickened but with only mild lumenal stenosis. Mitral and aortic valve disease was also present. There is one previous report of ventricular aneurysm in Hunter's syndrome. Pathophysiological contributions to aneurysm formation may include abnormal coronary flow, the presence of aortic stenosis and abnormal myocardium. This patient's sudden deterioration after a long period of clinical stability reinforces the need for careful follow-up of patients with cardiac manifestations of Hunter's disease. PMID- 11934601 TI - Changes in heat shock protein 70 localization and its content in rabbit aorta at various stages of experimental atherosclerosis. AB - Heat shock proteins, important components of cellular defense, are involved in the process of atherosclerosis. The present study aimed to evaluate the changes in distribution of hsp70 and its aortic content at various stages of high fat diet-induced atherosclerosis in the rabbits. Rabbits were fed on HFD (0.5% cholesterol+10% table butter) for 1, 3 and 6 months and hsp70 expression analyzed in normal and atherosclerotic aortae. Normal and 1 month group aortae showed normal histology and a homogeneous distribution of hsp70 while aortae from 3 and 6 months group showed atherosclerotic lesions and a heterogeneous distribution of hsp70. Immunoblot assay revealed an increase in aortic hsp70 levels on HFD feeding for 3 and 6 months. The possible role of elevated levels of hsp70 during atherosclerosis is discussed. PMID- 11934602 TI - Ultrastructure of intramural coronary arteries in pigs with hypertrophic cardiomyopathy. AB - Pigs with hypertrophic cardiomyopathy diagnosed by echocardiographic examination were selected for study from a genetic breeding herd. Under dissecting microscopic examination, intramural coronary arteries in the septum and left ventricular free wall of euthanized pigs were collected for ultrastructural study. The major lesions of wall thickening included degeneration or denudation of endothelium, subendothelial edema, proliferation of collagen fiber, and hyperplasia of smooth muscle cells. Smooth muscle cells proliferated and migrated through the internal elastic lamella into the intima, which caused the early lesion of wall thickening of the intramural coronary arteries. The extent of smooth muscle cell proliferation was related to the severity of endothelial damage. The smooth muscle cells in the intima were identified by immunohistochemical staining (i.e., smooth muscle actin [SMA] stain). Three major types of severe wall thickening with narrow lumen were observed in the intramural coronary arteries. Edema in the intima caused the major lesion of Type I wall thickening. The internal elastic lamella was broken into small interrupted fragments, and fine fragments of elastic fibers surrounded by the cellular processes of smooth muscle were observed in Type I lesions. Many smooth muscle cells proliferated in the intima and media, which constituted the major lesion of Type II wall thickening of the intramural coronary arteries. Many vacuolized, degenerated smooth muscle cells with fewer sarcoplasmic myofilaments could be clearly observed in the Type II lesions. In advanced cases, severe vacuolization and degeneration of smooth muscle cells with the presence of many bizarrely shaped smooth muscle cells in the walls of the intramural coronary arteries could be observed, which caused the major lesion of Type III wall thickening. Pigs with hypertrophic cardiomyopathy, characterized by spontaneously occurring lesions in intramural coronary arteries, may prove a valuable animal model for human disease. PMID- 11934603 TI - Pathophysiology of viral myocarditis: the role of humoral immune response. AB - The pathophysiology of viral myocarditis is still a matter of debate. Humoral autoimmunity in postviral heart disease remains an attractive but complex hypothesis. Antigenic mimicry with or without cytolytic antibody properties has been shown to play a role in the immunopathogenesis of myocarditis with respect to sarcolemmal/myolemmal epitopes (including the beta-receptor), myosin and some mitochondrial proteins including the antinucleotide translocator (ANT)-carrier and dihydrolipoamid dehydrogenase. Today, refined two-dimensional Western blots are able to identify receptors and enzymes that are target of a humoral immune response or the consequence of an "immunization process." A humoral immune response to an invading agent will most likely lead to immunodestruction first. After conversion to IgG, the continuing antibody response may indicate the healing or healed process and last for many years or life-long. This paper reviews our present knowledge on the humoral immune response in myocarditis and its interplay with the viral agents and the other components of the immune system. PMID- 11934604 TI - Tree barking? Not always an aortic prerogative. PMID- 11934608 TI - Structure/function relationships in OmpR and other winged-helix transcription factors. AB - Response regulators are the output component of two-component regulatory systems, the predominant form of signal transduction systems utilized by prokaryotes. The majority of response regulators function as transcription factors, yet detailed descriptions of their mechanisms of DNA binding and its consequences are lacking. Versatility in the modes of DNA binding is evident with winged helix-turn-helix proteins, raising doubts that mechanisms of DNA binding will be generalizable among members of the family. The current focus of some of the research efforts aimed at understanding activation and DNA binding by response regulators is highlighted in this review. PMID- 11934610 TI - Mining genome databases to identify and understand new gene regulatory systems. AB - The availability of a large number of sequenced microbial genomes allows us to conduct systematic studies on microbial gene regulatory systems. Computational methods, using comparative genomics approaches, are powerful tools to understand their mechanisms and evolutionary history. Recent advances in computational methodology for uncovering transcriptional regulatory components and their interactions are discussed. PMID- 11934609 TI - Molecular recognition of bacterial phosphorelay proteins. AB - The transfer of the phosphoryl group from a histidine kinase to a response regulator forms the basis of bacterial signal transduction. The critical question of how a component of a signal transduction system specifically associates with its partner to produce the ideal environment for phosphotransfer is addressed in this review in the light of the structure of the Spo0F-Spo0B complex in Bacillus subtilis. PMID- 11934611 TI - The biological roles of trans-translation. AB - The unique transfer-messenger RNA (tmRNA) molecule has been identified in all bacterial species examined, suggesting that its action confers an important survival advantage to bacteria. Acting both as a tRNA and an mRNA, in a process known as trans-translation, tmRNA adds a short peptide tag to undesirable proteins. Trans-translation plays at least two physiological roles: removing ribosomes stalled upon mRNA, and targeting the resulting truncated proteins for degradation by proteases. The first of these roles is required for all known activities of tmRNA, whereas the second may be dispensed with in most cases with little biological effect. However, tmRNA-targeted proteolysis may be important for fine-tuning expression of certain genes by altering the concentration of regulatory proteins. Here, we review recent literature that addresses the biological functions of tmRNA. PMID- 11934612 TI - Regulation of flagellar assembly. AB - Research on the molecular mechanism of bacterial flagellar assembly has been an ongoing study that spans three decades. Early work showed that regulation of flagellar gene transcription was coupled to the assembly process. Recent advances in the understanding of the regulation of flagellar assembly have shown that translational and post-translational regulation also plays a significant role in flagellar assembly. In both Salmonella and Caulobacter crescentus, translational regulation influences the secretion of the anti-sigma(28) factor FlgM and the flagellin fljK, respectively. Post-translational regulatory mechanisms also control the length of the hook and the ability of the type III secretion system to discriminate between middle and late secretion substrates. The flagellum provides a model system for understanding how gene regulation functions to ensure the efficient assembly of a complex structure and fundamental mechanisms common to all type III secretion systems. PMID- 11934613 TI - Regulation of type III secretion systems. AB - Type III secretion systems are utilised by numerous Gram-negative bacteria to efficiently interact with a host. Appropriate expression of type III genes is achieved through the integration of several regulatory pathways that ultimately co-ordinate the activity of a central transcriptional activator usually belonging to the AraC family. The complex regulatory cascades allow this virulence strategy to be utilised by different bacteria even if they occupy diverse niches that define a unique set of environmental cues. Simulating the appropriate combination of signals in vitro to allow a meaningful interpretation of the type III assembly and secretion regulatory cascade remains a common goal for researchers. Pieces of the puzzle slowly emerge to provide insightful views into the complex regulatory networks that allow bacteria to assemble and utilise type III secretion to efficiently colonise a host. PMID- 11934614 TI - Transcriptional, chemosensory and cell-contact-dependent regulation of type IV pilus expression. AB - Expression of Type IV pili (Tfp), multifunctional surface appendages expressed by Gram-negative species of medical and environmental significance, has previously been shown to be regulated by consensus two-component systems. Elucidation of their unique biogenesis pathway and the dynamics of pilus growth and retraction involved in motility have revealed a novel step at which regulation might be imposed. Studies of Tfp expression following adherence to host tissue clearly demonstrate regulation by modulation of the retraction process. In addition, a large set of components related to flagellar chemosensory pathways has been shown to influence Tfp expression levels in many species. Like their flagellar counterparts, the Tfp-dedicated homologues are proposed to function by regulating motor function. Rather than dictating the switch frequencies of organelle rotation, however, they are hypothesized to control the rates of fiber extrusion and retraction. PMID- 11934615 TI - Bud-site selection and cell polarity in budding yeast. AB - Polarized growth involves a hierarchy of events such as selection of the growth site, polarization of the cytoskeleton to the selected growth site, and transport of secretory vesicles containing components required for growth. The budding yeast Saccharomyces cerevisiae is an excellent model system for the study of polarized cell growth. A large number of proteins have been found to be involved in these processes, although their mechanisms of action are not yet well understood. Recent discoveries have helped elucidate many of the processes involved in cell polarity and bud-site selection in yeast and have modified the traditional view of cellular structures involved in these processes. This review focuses on recent advances on the roles of cortical tags, GTPases and the cytoskeleton in the generation and maintenance of cell polarity in yeast. PMID- 11934616 TI - Regulation of gene expression in the PTS in Escherichia coli: the role and interactions of Mlc. AB - Mlc represses several glucose-related genes, including the phosphotransferase system (PTS) genes ptsHI and ptsG. Induction of these genes by glucose occurs as a response to the flux of glucose through the PTS and involves the sequestration of Mlc to membranes containing dephosphorylated PtsG. In addition, ptsG levels are post-transcriptionally regulated by the glycolytic flux. PtsG is, thus, subject to multiple layers of control that, in turn, feed back on the regulation of the PTS by Mlc. The regulatory function of Mlc is compared with that of FruR and of the PTS-regulation-domain-containing protein, BglG. PMID- 11934617 TI - Active transport of iron and siderophore antibiotics. AB - Bacteria solubilize iron (Fe(3+)) with secreted siderophores, which are then taken up as Fe(3+)-siderophore complexes. Some bacteria also use iron in heme, hemoglobin, hemopexin, transferrin and lactoferrin of eukaryotic hosts. Crystal structures of two outer membrane transport proteins, FhuA and FepA, and biochemical data reveal strong long-range conformational changes of the proteins upon binding of Fe(3+)-siderophore complexes and in response to energy transfer from the cytoplasmic membrane into the outer membrane via the TonB-ExbB-ExbD protein complex. The crystal structure of the periplasmic binding protein FhuD strongly deviates from the uniform overall structure of binding proteins hitherto determined. Sideromycins, antibiotics that contain Fe(3+)-siderophore complexes as carriers, are highly effective, as they enter cells via Fe(3+)-siderophore transport systems. In this review, recently published data is discussed to demonstrate the state of understanding of iron transport across the outer membrane and the cytoplasmic membrane. PMID- 11934618 TI - Phospholipid modifications in bacteria. AB - Many bacteria alter the acyl chains of their membrane phospholipids in response to changing environmental conditions. Two of these modifications are reviewed: cis-->trans isomerization and cyclopropanation of double bonds. The isomerization reaction is now known to be catalyzed by a periplasmic protein that contains a covalently bound heme. Cyclopropanation has been shown to play a role in Mycobacterium tuberculosis pathogenesis and, in Escherichia coli, plays an important role in resistance to acidic conditions. PMID- 11934619 TI - Regulation of the desaturation of fatty acids and its role in tolerance to cold and salt stress. AB - The expression of cold-inducible genes is regulated by a two-component system in Synechocystis and Bacillus subtilis. The cold sensors are membrane-bound histidine kinases and it seems likely that they sense and transduce changes in the fluidity of membranes. Desaturation of fatty acids in membrane lipids has been implicated in tolerance to cold and salt stress. PMID- 11934620 TI - Connections between transcriptional regulation and type III secretion? AB - Type III secretion is a mechanism of protein export in bacteria, by which proteins (termed effectors) are exported from the bacteria to the external environment. One topic that has escaped much attention is the potential link between the secretion of effectors and transcriptional regulation of genes encoding the effectors. The goal of this commentary is to highlight some of the work that has been done and to stimulate additional research. PMID- 11934621 TI - Bacterial cell-to-cell communication: sorry, can't talk now - gone to lunch! AB - The identification of novel bacterial cell-to-cell communication (quorum sensing) systems based on diffusible signal molecules, such as indole and the LuxS autoinducer-2, requires discrimination between true signalling molecules and metabolites present in culture supernatants. This depends on rigorous chemical characterisation and demonstration that the molecule controls cellular responses beyond those required to metabolise or detoxify the signal. PMID- 11934642 TI - Osteoclastic acidification pathways during bone resorption. AB - Osteoclasts resorb bone by attaching to the surface and then secreting protons into an extracellular compartment formed between osteoclast and bone surface. This secretion is necessary for bone mineral solubilization and the digestion of organic bone matrix by acid proteases. This study summarizes the characterization and role of each type of ion transport and defines the main biochemical mechanisms involved in the dissolution of bone mineral during bone resorption. The primary mechanism responsible for acidification of the osteoclast-bone interface is vacuolar H+-adenosine triphosphatase (ATPase) coupled with Cl- conductance localized to the ruffled membrane. Carbonic anhydrase II (CAII) provides the proton source for extracellular acidification by H+-ATPase and the HCO3- source for the HCO3-/Cl- exchanger. Whereas some transporters are responsible for the bone resorption process, others are essential for pH regulation in the osteoclast. The HCO3-/Cl- exchanger, in association with CAII, is the major transporter for maintenance of normal intracellular pH. An Na+/H+ antiporter may also contribute to the recovery of intracellular pH during early osteoclast activation. Once this mechanism has been rendered inoperative, another conductive pathway translocates the protons and modulates cytoplasmic pH. Inward rectifying K+ channels may also be involved by compensating for the external acidification due to H+ transport. These different effects of transport processes, either on bone resorption or pH homeostasis, increase the number of possible sites for pharmacological intervention in the treatment of metabolic bone diseases. PMID- 11934643 TI - Mechanical properties of skeletal bone in gene-mutated stopsel(dtl28d) and wild type zebrafish (Danio rerio) measured by atomic force microscopy-based nanoindentation. AB - An atomic force microscopy (AFM)-based nanoindenter was used to evaluate the mechanical properties of skeletal bones in wild-type and gene-mutated zebrafish (Danio rerio), stopsel(dtl28d). Both skeletons were isolated from adult zebrafish and tested under a load of 5 mN. It was found that stp/stp bone has a similar nanohardness but significantly greater elastic modulus compared with that of wild type bone. The residual indenter impressions using AFM and the fracture surfaces of both bones using scanning electron microscopy were examined and showed that the bone of zebrafish becomes more brittle after the stp mutation. This first observation of the alteration of bone mechanical behavior by gene mutation in zebrafish system is of scientific and clinical relevance to many areas of study, such as bone fracture and fragility mechanisms in human heritable disorders and bone-materials fabrication via gene engineering. PMID- 11934644 TI - Signaling by mechanical strain involves transcriptional regulation of proinflammatory genes in human periodontal ligament cells in vitro. AB - Intracellular signals generated by mechanical strain profoundly affect the metabolic function of osteoblast-like periodontal ligament (PDL) cells, which reside between the tooth and alveolar bone. In response to applied mechanical forces, PDL cells synthesize bone-resorptive cytokines to induce bone resorption at sites exposed to compressive forces and deposit bone at sites exposed to tensile forces in an environment primed for catabolic processes. The intracellular mechanisms that regulate this bone remodeling remain unclear. Here, in an in vitro model system, we show that tensile strain is a critical determinant of PDL-cell metabolic functions. Equibiaxial tensile strain (TENS), when applied at low magnitudes, acts as a potent antagonist of interleukin (IL) 1beta actions and suppresses transcriptional regulation of multiple proinflammatory genes. This is evidenced by the fact that TENS at low magnitude: (i) inhibits recombinant human (rh)IL-1beta-dependent induction of cyclooxygenase 2 (COX-2) mRNA expression and production of prostaglandin estradiol (PGE2); (ii) inhibits rhIL-1beta-dependent induction matrix metalloproteinase-1 (MMP-1) and MMP-3 synthesis by suppressing their mRNA expression; (iii) abrogates rhIL-1beta induced suppression of tissue inhibitor of metalloprotease-II (TIMP-II) expression; and (iv) reverses IL-1beta-dependent suppression of osteocalcin and alkaline phosphatase synthesis. Nevertheless, these actions of TENS were observed only in the presence of IL-1beta, as TENS alone failed to affect any of the aforementioned responses. The present findings are the first to show that intracellular signals generated by low-magnitude mechanical strain interfere with one or more critical step(s) in the signal transduction cascade of rhIL-1beta upstream of mRNA expression, while concurrently promoting the expression of osteogenic proteins in PDL cells. PMID- 11934645 TI - Vector-averaged gravity regulates gene expression of receptor activator of NF kappaB (RANK) ligand and osteoprotegerin in bone marrow stromal cells via cyclic AMP/protein kinase A pathway. AB - Bone loss due to unloading of the skeleton may be caused by an acceleration of osteoclastic bone resorption as well as a decline of osteoblastic bone formation. Recently, two molecular species that play important roles in osteoclastogenesis were discovered: (i) the receptor activator of NF-kappaB ligand (RANKL)/osteoprotegerin (OPG) ligand/osteoclast differentiation factor induces osteoclastogenesis; and (ii) the OPG/osteoclastogenesis inhibitory factor potently inhibits osteoclastogenesis. To investigate the effects of gravity on gene expression of RANKL and OPG, a mouse bone marrow-derived stromal cell line, ST2, was cultured on a single axis clinostat, which generates a vector-averaged gravity environment. Northern blot analysis revealed that RANKL mRNA was increased, whereas that of OPG decreased. The clinostat culture also caused an increase in intracellular cyclic (cAMP) level. Both forskolin and dibutyryl-cAMP mimicked the regulation of RANKL and OPG transcription in clinostat culture. These modulations of gene expression in clinostat culture were blocked by a protein kinase A (PKA) inhibitor, H89, but not by a cyclooxygenase inhibitor, indomethacin. The enhancement of RANKL gene expression under clinostat culture and its inhibition by H89 were confirmed by a reporter assay with the murine RANKL 5'-flanking region. These results suggest that modulations of RANKL and OPG expression in stromal cells might be one of the causes of bone loss during skeletal unloading. An elevation of intracellular cAMP level caused through an as yet undetermined pathway is involved in modulation of RANKL and OPG expression during clinostat culture. PMID- 11934646 TI - Calcium-sensing receptor-mediated activation of phospholipase C-gamma1 is downstream of phospholipase C-beta and protein kinase C in MC3T3-E1 osteoblasts. AB - Elevated extracellular calcium (Ca(e)) stimulates both chemotaxis and mitogenesis of MC3T3-E1 osteoblasts via a calcium-sensing receptor (CasR). Ca(e)-mediated chemotaxis of these bone-forming cells is dependent on phospholipase C (PLC) and blocked by the Gi-protein inhibitor pertussis toxin. In this study, we examine the signaling mechanisms by which the CasR stimulates PLC activity in MC3T3-E1 osteoblasts. We found that elevated Ca(e) stimulated PLC-gamma1 tyrosine phosphorylation in a time-dependent and Ca(e)-concentration-dependent manner. The maximal increase in PLC-gamma1 tyrosine phosphorylation was observed 3-5 min after increasing Ca(e) by 3.2 mmol/L from 1.8 mmol/L. Elevated Ca(e) also promoted a rapid increase in both inositol 1,4,5-trisphosphate [Ins(1,4,5)P3], a second messenger formed by PLC-mediated hydrolysis of phosphatidylinositol 4,5 bisphosphate, and cytosolic free calcium ([Ca+2]i). The kinetics of the CasR mediated increases in Ins(1,4,5)P3 and [Ca+2]i and the sensitivity of the Ca(e) stimulated elevation in [Ca+2]i to U73122 (a PLC inhibitor) together suggest that the osteoblast CasR is coupled via Gq to PLC-beta. U73122 blocked the Ca(e) promoted, but not PDGF-promoted, PLC-gamma1 tyrosine phosphorylation, suggesting that the activation of PLC-beta is upstream of PLC-gamma1 activation. Inhibition of protein kinase C (PKC) disrupted Ca(e)-stimulated tyrosine phosphorylation of PLC-gamma1. In addition, exposure to pertussis toxin or exogenous activation of protein kinase A (PKA) inhibited PLC-gamma1 tyrosine phosphorylation in response to Ca(e). The results indicate that: (a) the osteoblast CasR activates PLC-gamma1 downstream of PLC-beta in a PKC-dependent manner; (b) PKA is a negative regulator of Ca(e)-promoted PLC-gamma1 phosphorylation; and (c) Gq and Gi are both involved in the CasR-mediated phosphorylation of PLC-gamma1. PMID- 11934648 TI - Volume increase in growth plate chondrocytes during hypertrophy: the contribution of organic osmolytes. AB - During the differentiation cascade of growth plate chondrocytes, cells undergo as much as a 10-15-fold increase in volume. This volume increase, which occurs to different extents in growth plates growing at different rates, has been demonstrated to be the single most significant variable in understanding the quantitative aspects of the cellular kinetics of long bone growth. Our hypothesis is that this volume increase, which occurs through cell swelling by water imbibition, requires intracellular accumulation of osmolytes through activation or upregulation of membrane transport mechanisms. Significant intracellular accumulation of inorganic osmolytes, such as Na+, K+, and Cl-, is potentially disruptive to normal cellular metabolism, whereas intracellular accumulation of organic osmolytes is considered to be more compatible with metabolic function. Thus, we concentrated on determining the contributions of organic osmolytes- betaine, amino acids, inositol, and sorbitol--to volume increase. Pooled cryostat sections of young bovine growth plates were extracted followed by automated analysis for their content of amino acids. Analysis for betaine and the sugar alcohols was done by extraction and derivatization, followed by high-performance liquid chromatography (HPLC). Parallel stereological analyses correlated osmolyte changes to stages of chondrocytic differentiation, specifically comparing intracellular concentration and amount in proliferative vs. hypertrophic chondrocytes. Calculations demonstrated that, maximally, these organic osmolytes, in total, account for 6%-7% of the intracellular osmolytes required to sustain the volume increase, and that the most significant contribution is from betaine. This suggests that intracellular accumulation of organic osmolytes is not a primary strategy used by growth plate chondrocytes during volume increase of their terminal differentiation. The data also suggest that there is a differential regulation of transporters of these osmolytes such that intracellular concentrations are constantly modified as cells proceed through the differentiation cascade. PMID- 11934647 TI - Effects of prostaglandin E2 on gene expression in primary osteoblastic cells from prostaglandin receptor knockout mice. AB - Recent studies have shown that stimulation of osteoclastogenesis in cocultures of osteoblasts and spleen cells in response to prostaglandin E2 (PGE2) is markedly decreased when the osteoblasts are derived from cells lacking either the EP2 or the EP4 receptor. Induction of osteoclast formation requires upregulation of receptor activator of nuclear factor-kappaB ligand (RANKL) on cells of the osteoblastic lineage, which then binds to the RANK receptor on cells of the osteoclast lineage. Osteoprotegerin (OPG) is a decoy receptor for RANKL that can block its interaction with RANK. In addition, macrophage-colony stimulating factor (M-CSF) is essential for osteoclast formation. Finally, PGE2 can increase interleukin-6 (IL-6), which may further enhance osteoclastogenesis. To study the relative influence of the EP2 and EP4 receptors on response of these factors to PGE2, we examined mRNA levels for RANKL, OPG, M-CSF, and IL-6 in primary osteoblastic cell cultures derived from two lines of EP2 knockout mice (EP2-/-) and one line of EP4 knockout mice (EP4-/-) and the relevant wild-type controls (EP2+/+ and EP4+/+). The responses of cells from wild-type animals of all three lines were similar. After PGE2 treatment, RANKL mRNA levels were increased at 2 h, and this was sustained over 72 h. Basal RANKL expression was moderately reduced in EP2-/- cells and markedly reduced in EP4-/- cells. PGE2 increased RANKL mRNA in EP2-/- cells and EP4-/- cells, but the levels were significantly reduced compared with wild-type cells. There were no consistent changes in expression of M-CSF or OPG in the different genotypes or with PGE2 treatment. IL 6 mRNA was variably increased by PGE2 in both wild-type and knockout cells, although the absolute levels were somewhat lower in both EP2-/- and EP4 -/- cultures. Parathyroid hormone (PTH) increased RANKL and IL-6 and decreased OPG mRNA levels similarly in both wild-type and EP2-/- or EP4-/- cells. The major defect in the response to PGE2 in animals lacking either EP2 or EP4 receptors is a reduction in basal and stimulated RANKL levels. Loss of EP4 receptor appears to have a greater effect on basal RANKL expression than EP2. PMID- 11934649 TI - ED-71, a vitamin D analog, is a more potent inhibitor of bone resorption than alfacalcidol in an estrogen-deficient rat model of osteoporosis. AB - Although active vitamin D is used in certain countries for the treatment of osteoporosis, the risk of causing hypercalcemia/hypercalciuria means that there is only a narrow therapeutic window, and this has precluded worldwide approval. The results of our previous animal studies have suggested that the therapeutic effect of active vitamin D on bone loss after estrogen deficiency can be dissociated at least partly from its effect of enhancing intestinal calcium absorption and suppressing parathyroid hormone (PTH) secretion. To test this, we compared the effects of ED-71, a hydroxypropoxy derivative of 1alpha,25 dihydroxyvitamin D3, with orally administered alfacalcidol, on bone mineral density (BMD) and the bone remodeling process as a function of their effects on calcium metabolism and PTH, in a rat ovariectomy (ovx) model of osteoporosis. ED 71 increased bone mass at the lumbar vertebra to a greater extent than alfacalcidol, while enhancing calcium absorption (indicated by urinary calcium excretion) and decreasing serum PTH levels to the same degree as alfacalcidol. ED 71 lowered the biochemical and histological parameters of bone resorption more potently than alfacalcidol, while maintaining bone formation markers. These results suggest that active vitamin D exerts an antiosteoporotic effect by inhibiting osteoclastic bone resorption while maintaining osteoblastic function, and that these anticatabolic/anabolic effects of active vitamin D take place independently of its effects on calcium absorption and PTH. The demonstration that ED-71 is more potent in these properties than alfacalcidol makes it an attractive candidate as an antiosteoporotic drug. PMID- 11934650 TI - Effect of phosphodiesterase inhibitor-4, rolipram, on new bone formations by recombinant human bone morphogenetic protein-2. AB - Collagen sponge disks (6 mm diameter, 1 mm thickness) were impregnated with recombinant human bone morphogenetic protein-2 (rhBMP-2) (5 microg/disk) and implanted onto the back muscles of mice. Ten or 20 mg/kg per day of Rolipram, a selective inhibitory agent to phosphodiesterase type 4 (PDE-4), or vehicle, was injected subcutaneously into the host mice for 3 weeks. After treatment, rhBMP-2 induced ectopic ossicles were harvested and examined by radiographic and histologic methods to determine the size, bone quality, and mineral content of the ossicles. The ossicles from a group treated with 20 mg/kg per day Rolipram were significantly larger in size and higher in bone mineral density (BMD) and bone mineral content (BMC) than the control samples. No significant differences were noted in mice treated with 10 mg/kg per day of Rolipram. Histologically, ossicles from the high-dose (20 mg/kg per day) Rolipram-treated group showed densely packed, thicker trabeculae when compared with those from the control group. These experimental results indicate that the PDE-4 inhibitor, Rolipram, may enhance the bone-inducing capacity of BMP-2 in mesenchymal cells. This in turn may result in increased responsiveness to BMP-2 and point to a potential use of PDE-4 inhibitors for the promotion of rhBMP-dependent bone repair. PMID- 11934651 TI - Differences of three-dimensional trabecular microstructure in osteopenic rat models caused by ovariectomy and neurectomy. AB - We investigated the differences in three-dimensional microstructure of bone in cases of osteopenia caused by two different procedures: ovariectomy (ovx) and sciatic neurectomy (nx). Thirty-nine 8-week-old female Lewis rats were divided into two groups: (1) ovx and sham operation; and (2) nx and sham operation. At 12 weeks of age these rats were killed to sample the right tibiae. The samples were scanned using microcomputed tomography to obtain metric parameters such as bone volume fraction (BV/TV), trabecular thickness (Tb.Th), and trabecular separation (Tb.Sp), and nonmetric parameters such as structure model index (SMI), trabecular bone pattern factor (TBPf), and degree of anisotropy (DA). The changes in all microstructural parameters were significant in both the ovx and nx groups, with those of BV/TV, Tb.Th, Tb.Sp, and SMI more significant in the ovx group than in the nx group, in comparison to their respective controls. The significantly higher coefficient of variance for Tb.Th across the entire analyzed area of the individual samples indicated that the trabecular thinning occurred heterogenously and that the microstructural deterioration induced by ovx and nx appeared to be locally accelerated, so as to induce perforation and disappearance of trabeculae. The DA increased significantly in the ovx rats, whereas it decreased in the nx rats. The appearance of microstructural deterioration differed between the two osteopenic models. The three-dimensional (3D) images from the nx rats showed flake-like trabeculae, whereas the ovx rats exhibited a diffuse disappearance of trabeculae, especially in the central part of the tibia, but with a preservation of shape for those trabeculae that were retained. The reduction in cortical area was more significant in the nx group. nx and ovx resulted in significant changes in bone microstructure, showing perforation and removal of trabeculae due to locally accelerated bone resorption. The 3D microcomputed tomography images demonstrated the different microstructural changes that occurred in the ovx and nx groups. Loading during bone resorption increased the anisotropy, whereas immobilization increased the isotropy. In addition, immobilization had a more significant effect on the cortical area. PMID- 11934652 TI - Effect of 1 year of discontinuation of raloxifene or estrogen therapy on bone mineral density after 5 years of treatment in healthy postmenopausal women. AB - The beneficial effects of hormone replacement therapy (HRT), selective estrogen receptor modulators (SERMs), or bisphosphonates in the prevention and treatment of osteoporosis in postmenopausal women have been well established. However, little is known about the effects of discontinuation of treatment on bone mineral density. We investigated the effect of 1 year of discontinuation of the SERM raloxifene (Ral; 60 mg and 150 mg), conjugated equine estrogen (CEE; 0.625 mg), and placebo after 5 years of treatment in a double-blind, randomized study. Thirty-eight of 59 healthy and hysterectomized postmenopausal women (mean age 55 years) completed the treatment and 1 year follow-up period. Lumbar spine and femoral neck bone mineral density (BMD) were performed with dual-energy X-ray absorptiometry, before, during, and at the end of treatment, as well as after 1 year of discontinuation of therapy. One year of discontinuation significantly reduced the mean lumbar spine BMD in the raloxifene- and estrogen-treated women (p < 0.05), whereas mean femoral neck BMD was reduced significantly only in women treated with 60 mg Ral (p < 0.05). The mean percentage change (+/-SD) in lumbar spine BMD was: CEE, -6.2% (+/-3.7%); Ral 60 mg, -2.4% (+/-2.4%); Ral 150 mg, 2.6% (+/-3.1%); and placebo, -1.6% (+/-4.3%). Our results show that 5 years of treatment with either Ral or CEE did not protect against bone loss after 1 year of withdrawal of therapy, and that the rate of bone loss was not significantly different from that of placebo-treated women. PMID- 11934653 TI - Relationships between trabecular bone remodeling and bone vascularization: a quantitative study. AB - Beside its well-known role in bone development, vascularization plays a major role in bone cell migration for bone remodeling and metastatic tumor invasion. However, the various techniques used to identify vessels in bone have never been tested for trabecular bone vessel quantification, whereas bone remodeling quantitative parameters are commonly assessed. In this context, we developed and compared various histological techniques used to visualize blood vessels in rat bone in order to quantify them. First, several products were tested by intracardiac infusion to opacify the bone vascular network. The best results were obtained using either an India ink-1% agarose solution or an India ink-saturated barium sulfate solution followed by X-ray microradiography. Second, to identify the types of vessels, we also performed histoenzymology and immunohistochemistry stainings. Neither alkaline phosphatase (for endothelial cells) nor adenosine triphosphatase (ATPase) stainings (for smooth muscle cells) provided a low enough background to allow for vessel identification and quantification. For immunohistochemistry, various specific vessel constituents were analyzed: laminin, smooth muscle cell alpha-actin, factor VIII, and lectin Griffonia simplifolia. Anti-laminin and anti-smooth muscle cell alpha-actin antibodies gave the best results for quantification. Third, after optimization of these techniques, we performed quantitative bone and vessel histomorphometry on two groups of 12 rats each, for which bone remodeling and vessel number and area parameters were measured. No statistical differences were observed between the two groups, confirming the reproducibility of our measurements. A significant relationship was found between vessel number and histodynamic parameters; that is, bone formation rate correlated positively with India ink-positive vessel area (p < 0.009, r2 = 0.54) and alpha-actin-positive vessel number (p < 0.05, r2 = 0.66). Furthermore, we report reproducible techniques for visualization and quantification of vessels in bone that also allowed for simultaneous conventional bone histomorphometry. This methodology should help researchers to better understand the functional and anatomical relationship between trabecular bone and its vascularization during normal or pathological processes. PMID- 11934654 TI - Use of low potency estrogens does not reduce the risk of hip fracture. AB - High endogenous sexual hormone levels and use of medium potency estrogens are associated with a reduced risk of hip fracture in postmenopausal women. However, it is not clear if low potency estrogens confer the same benefits as the more widely used forms of menopausal hormone replacement. We examined the association between postmenopausal use of low potency estrogens, mainly estriol, and hip fracture risk in a population-based, case-control study. Using data from mailed questionnaires and telephone interviews, we analyzed the association between low potency estrogen use and hip fracture risk among 1327 cases, 50-81 years of age, and 3262 randomly selected age-matched controls. Ever use of low potency estrogens was reported by 19% of the cases and 23% of controls. Compared to with never users of any hormone replacement therapy, ever users of low potency estrogens had a multivariate odds ratio (OR) for hip fracture of 0.96 (95% confidence interval [CI] 0.67-1.39). Current use was also not associated with a reduction in risk: OR 0.94 (95% CI 0.58-1.53), and longer duration of use was also not associated with a risk reduction. Even current use of the highest dose of oral estriol (2 mg/day) conferred no risk reduction (OR 1.01, 95% CI 0.61 1.67) compared with never use of hormone replacement therapy. After exclusion of ever users of medium potency estrogens from the analyses, we found a risk reduction of fracture among current vaginal low potency estrogen users (multivariate OR 0.67, 95% CI 0.49-0.92). In contrast to medium potency estrogens, low potency estrogens did not confer a substantial overall reduction in hip fracture risk. PMID- 11934655 TI - Paradoxical adaptation of mature radius to unilateral use in tennis playing. AB - The positive effects of physical activity on human bone mass have been well documented in many cross-sectional studies comparing athletes with sedentary controls as well as in longitudinal follow-up. By applying peripheral quantitative computed tomography (pQCT), which has the advantage of measuring volumetric bone mineral density (BMD) and the ability to distinguish among trabecular and cortical components, it was demonstrated that cortical BMD of the dominant arm was not greater than that of the nondominant arm. Cortical drift toward the periosteal direction and an increase in cortical thickness resulted in an improvement of mechanical characteristics of the playing arm's midradius. An improvement in the mechanical properties of young adult bone in response to long term exercise was therefore related to geometric adaptation, but not to an increase in BMD. The manner in which the recruitment and function of bone cells are coordinated differs between the growing and the nongrowing skeleton. In the former, modeling is the dominant mode, and in the latter it is remodeling. In the present study, the side-to-side difference of 92 middle-aged female tennis players who initiated training after bone had matured was analyzed by pQCT. The side-to-side difference detected suggested a paradoxical adaptation of the mature radius to unilateral use during tennis playing, and that tennis playing after bone had matured did not stimulate cortical drift in the periosteal direction, unlike that seen in young subjects. Unexpectedly, the cross-sectional areas (periosteal and endocortical area) of the radius were smaller in the dominant arm than in the nondominant arm in the middle-aged female players. The findings suggest that unilateral use of the arm after the third decade of life suppresses age-related changes in bone geometry. PMID- 11934656 TI - Otosclerosis: a genetically heterogeneous disease involving at least three different genes. AB - Otosclerosis is caused by abnormal bone homeostasis of the otic capsule, resulting in hearing impairment in 0.3%-0.4% of the white population. The etiology of the disease remains unclear and environmental as well as genetic factors have been implicated. We localized the first autosomal-dominant locus to chromosome 15 in 1998 (OTSC1) in an Indian family and, recently, we reported the localization of a second gene for otosclerosis to a 16 cM interval on chromosome 7q (OTSC2). In this study, we recruited and analyzed nine additional families (seven Belgian and two Dutch families with 53 affected and 20 unaffected subjects) to investigate the importance of these loci in autosomal-dominant otosclerosis. We completed linkage analysis with three microsatellite markers of chromosome 15 (D15S652, D15S1004, D15S657) and five microsatellite markers of chromosome 7 (D7S495, D7S2560, D7S684, D7S2513, D7S2426). In two families, results compatible with linkage to OTSC2 were found, but in the seven remaining families OTSC1 and OTSC2 were excluded. Heterogeneity testing provided significant evidence for genetic heterogeneity, with an estimated 25% of families linked to OTSC2. These results indicate that, besides OTSC1 and OTSC2, there must be at least one additional otosclerosis locus. PMID- 11934657 TI - The cost of osteoporosis in men: the French situation. AB - The objective of this study was to estimate the annual direct medical costs of osteoporosis and osteoporotic fractures incurred by French men > or =50 years of age. Costs were assessed from a societal perspective for 1999 and expressed in Euros. An expert panel was consulted to identify fractures attributable to osteoporosis according to International Classification of Diseases (ICD)-10 codes. Available age- and gender-specific osteoporosis attribution probabilities (OAP) were used to derive the proportion of health-care utilization for fractures resulting from osteoporosis. Hospital and ambulatory care costs due to fractures were obtained from French databases. A total of 23,260 acute hospitalizations were found to be caused by osteoporosis, 52% of which are for hip fractures. Mean cost per stay varied widely according to the site of fracture, from 1300 (wrist fracture) to 5900 (hip fracture). Consequently, the total cost of acute hospitalization amounts to 97.6 million, with hip fractures accounting for 73.2% of the expenditures. Rehabilitation and convalescence costs were estimated to be 90.8 million, generating a total hospital cost of 188.4 million. The total outpatient costs were estimated to be 9.1 million. Thus, the total medical costs of male osteoporosis amount to 197.5 million. A sensitivity analysis was performed to test the robustness of this figure. We estimate the number of fractures by applying international incidence rates from the literature to the French male population aged > or =50 years, whereas the OAP and unit costs were kept constant. This approach yielded an estimate of 21,857 fractures, which is only 6% below the base case. When compared with values from other countries, our study results appear very conservative. PMID- 11934658 TI - Number of years since menopause: spontaneous bone loss is dependent but response to hormone replacement therapy is independent. AB - In this study we examine the influence of number of years since menopause on spontaneous bone loss and response to hormone replacement therapy (HRT) in 274 women (56.1 +/- 4.2 years) completing two placebo-controlled HRT studies of 2 or 3 year duration. Both cross sectionally and longitudinally, bone loss in untreated women was greatest closest to menopause and declined thereafter (r = 0.34, p < 0.01 for lumbar spine bone loss and r = 0.25, p < 0.05 for femoral neck bone loss when correlated with number of years since menopause), such that the loss was eliminated in the femoral neck and bone mass increased in the spine in women >10 years after menopause. In contrast, bone turnover was consistently elevated throughout postmenopause, both cross-sectionally and longitudinally. The association with number of years since menopause was counteracted by both 1 and 2 mg estradiol combined with gestodene, piperazine, estrone sulfate in combination with norethisterone, and a combination of 2 mg estradiol and 1 mg norethisterone acetate. In addition, the response to various HRT regimens was independent of baseline bone mass. Whereas bone loss was significantly related to number of years since menopause, all HRT regimens applied arrested bone loss in healthy postmenopausal women, regardless of number of years since menopause. PMID- 11934659 TI - Bone mineral density and mortality in elderly men and women: the Rotterdam Study. AB - Recent studies have shown that a low bone mineral density (BMD) is associated with a higher risk of mortality. Most studies have investigated this relationship in women only and presented their risk estimates per standard deviation change in BMD. However, when using this approach, a BMD threshold might be missed when relative risks are presented in the traditional manner. Therefore, in this study our aim was to model the relation between BMD and all-cause mortality. In the Rotterdam Study, follow-up was complete for 5819 men and women aged > or =55 years for whom BMD data were available. During an average follow-up of 5.4 years, 399 men and 317 women died. We calculated BMD Z scores using measurements performed at the femoral neck. Cox proportional hazards regression was used to fit the model. An average BMD, reflected by a Z score = 0, was used as the reference. For women, no significant relationship between BMD and overall mortality was observed. For men, however, a cubic model best fitted the relationship under study, also after adjusting for age and body mass index (BMI). The risk of mortality increased when BMD was below average. Similar results were found when separate curves were made for diabetics and nondiabetics, smokers (ever or never), and tertiles of BMI. Excluding subjects who had suffered hip fractures, or adjusting for the number of drugs used and for lower limb disability, essentially did not change results. This suggests that low BMD is not mainly due to morbidity and impaired mobility in our cohort, which makes this a less likely explanation for the observed relation with mortality. The results of our study suggest that, in men, a nonlinear relationship between BMD and mortality exists, which is independent of comorbidity, whereas, in women, no significant relationship was observed. PMID- 11934660 TI - Evidence of geographic variation in the occurrence of Paget's disease. PMID- 11934661 TI - Mouse models of insulin resistance. AB - The hallmarks of type 2 diabetes are impaired insulin action in peripheral tissues and decreased pancreatic beta-cell function. Classically, the two defects have been viewed as separate entities, with insulin resistance arising primarily from impaired insulin-dependent glucose uptake in skeletal muscle, and beta-cell dysfunction arising from impaired coupling of glucose sensing to insulin secretion. Targeted mutagenesis and transgenesis involving components of the insulin action pathway have changed our understanding of these phenomena. It appears that the role of insulin signaling in the pathogenesis of type 2 diabetes has been overestimated in classic insulin target tissues, such as skeletal muscle, whereas it has been overlooked in liver, pancreatic beta-cells, and brain, which had been thought not to be primary insulin targets. We review recent progress and try to reconcile areas of apparent controversy surrounding insulin signaling in skeletal muscle and pancreatic beta-cells. PMID- 11934662 TI - Contribution of the endoplasmic reticulum to the glucose-induced [Ca(2+)](c) response in mouse pancreatic islets. AB - Thapsigargin (TG), a blocker of Ca(2+) uptake by the endoplasmic reticulum (ER), was used to evaluate the contribution of the organelle to the oscillations of cytosolic Ca(2+) concentration ([Ca(2+)](c)) induced by repetitive Ca(2+) influx in mouse pancreatic beta-cells. Because TG depolarized the plasma membrane in the presence of glucose alone, extracellular K(+) was alternated between 10 and 30 mM in the presence of diazoxide to impose membrane potential (MP) oscillations. In control islets, pulses of K(+), mimicking regular MP oscillations elicited by 10 mM glucose, induced [Ca(2+)](c) oscillations whose nadir remained higher than basal [Ca(2+)](c). Increasing the depolarization phase of the pulses while keeping their frequency constant (to mimic the effects of a further rise of the glucose concentration on MP) caused an upward shift of the nadir of [Ca(2+)](c) oscillations that was reproduced by raising extracellular Ca(2+) (to increase Ca(2+) influx) without changing the pulse protocol. In TG-pretreated islets, the imposed [Ca(2+)](c) oscillations were of much larger amplitude than in control islets and occurred on basal levels. During intermittent trains of depolarizations, control islets displayed mixed [Ca(2+)](c) oscillations characterized by a summation of fast oscillations on top of slow ones, whereas no progressive summation of the fast oscillations was observed in TG-pretreated islets. In conclusion, the buffering capacity of the ER in pancreatic beta-cells limits the amplitude of [Ca(2+)](c) oscillations and may explain how the nadir between oscillations remains above baseline during regular oscillations or gradually increases during mixed [Ca(2+)](c) oscillations, two types of response observed during glucose stimulation. PMID- 11934663 TI - Partitioning glucose distribution/transport, disposal, and endogenous production during IVGTT. AB - We have separated the effect of insulin on glucose distribution/transport, glucose disposal, and endogenous production (EGP) during an intravenous glucose tolerance test (IVGTT) by use of a dual-tracer dilution methodology. Six healthy lean male subjects (age 33 +/- 3 yr, body mass index 22.7 +/- 0.6 kg/m(2)) underwent a 4-h IVGTT (0.3 g/kg glucose enriched with 3-6% D-[U-(13)C]glucose and 5-10% 3-O-methyl-D-glucose) preceded by a 2-h investigation under basal conditions (5 mg/kg of D-[U-(13)C]glucose and 8 mg/kg of 3-O-methyl-D-glucose). A new model described the kinetics of the two glucose tracers and native glucose with the use of a two-compartment structure for glucose and a one-compartment structure for insulin effects. Insulin sensitivities of distribution/transport, disposal, and EGP were similar (11.5 +/- 3.8 vs. 10.4 +/- 3.9 vs. 11.1 +/- 2.7 x 10(-2) ml small middle dot kg(-1) small middle dot min(-1) per mU/l; P = nonsignificant, ANOVA). When expressed in terms of ability to lower glucose concentration, stimulation of disposal and stimulation of distribution/transport accounted each independently for 25 and 30%, respectively, of the overall effect. Suppression of EGP was more effective (P < 0.01, ANOVA) and accounted for 50% of the overall effect. EGP was suppressed by 70% (52-82%) (95% confidence interval relative to basal) within 60 min of the IVGTT; glucose distribution/transport was least responsive to insulin and was maximally activated by 62% (34-96%) above basal at 80 min compared with maximum 279% (116-565%) activation of glucose disposal at 20 min. The deactivation of glucose distribution/transport was slower than that of glucose disposal and EGP (P < 0.02) with half-times of 207 (84-510), 12 (7-22), and 29 (16-54) min, respectively. The minimal-model insulin sensitivity was tightly correlated with and linearly related to sensitivity of EGP (r = 0.96, P < 0.005) and correlated positively but nonsignificantly with distribution/transport sensitivity (r = 0.73, P = 0.10) and disposal sensitivity (r = 0.55, P = 0.26). We conclude that, in healthy subjects during an IVGTT, the two peripheral insulin effects account jointly for approximately one-half of the overall insulin-stimulated glucose lowering, each effect contributing equally. Suppression of EGP matches the effect in the periphery. PMID- 11934664 TI - Regulation of GLUT4 biogenesis in muscle: evidence for involvement of AMPK and Ca(2+). AB - There is evidence suggesting that adaptive increases in GLUT4 and mitochondria in skeletal muscle occur in parallel. It has been reported that raising cytosolic Ca(2+) in myocytes induces increases in mitochondrial enzymes. In this study, we tested the hypothesis that an increase in cytosolic Ca(2+) induces an increase in GLUT4. We found that raising cytosolic Ca(2+) by exposing L6 myotubes to 5 mM caffeine for 3 h/day for 5 days induced increases in GLUT4 protein and in myocyte enhancer factor (MEF)2A and MEF2D, which are transcription factors involved in regulating GLUT4 expression. The caffeine-induced increases in GLUT4 and MEF2A and MEF2D were partially blocked by dantrolene, an inhibitor of sarcoplasmic reticulum Ca(2+) release, and completely blocked by KN93, an inhibitor of Ca(2+) calmodulin-dependent protein kinase (CAMK). Caffeine also induced increases in MEF2A, MEF2D, and GLUT4 in rat epitrochlearis muscles incubated with caffeine in culture medium. 5-Aminoimidazole-4-carboxamide ribonucleoside (AICAR), which activates AMP-activated protein kinase (AMPK), also induced approximately twofold increases in GLUT4, MEF2A, and MEF2D in L6 myocytes. Our results provide evidence that increases in cytosolic Ca(2+) and activation of AMPK, both of which occur in exercising muscle, increase GLUT4 protein in myocytes and skeletal muscle. The data suggest that this effect of Ca(2+) is mediated by activation of CAMK and indicate that MEF2A and MEF2D are involved in this adaptive response. PMID- 11934665 TI - Evidence of a malonyl-CoA-insensitive carnitine palmitoyltransferase I activity in red skeletal muscle. AB - Carnitine palmitoyltransferase I (CPT I), which is expressed as two distinct isoforms in liver (alpha) and muscle (beta), catalyzes the rate-limiting step in the transport of fatty acid into the mitochondria. Malonyl-CoA, a potent inhibitor of CPT I, is considered a key regulator of fatty acid oxidation in both tissues. Still unanswered is how muscle beta-oxidation proceeds despite malonyl CoA concentrations that exceed the IC(50) for CPT Ibeta. We evaluated malonyl-CoA suppressible [(14)C]palmitate oxidation and CPT I activity in homogenates of red (RG) and white (WG) gastrocnemius, soleus (SOL), and extensor digitorum longus (EDL) muscles. Adding 10 microM malonyl-CoA inhibited palmitate oxidation by 29, 39, 60, and 89% in RG, SOL, EDL, and WG, respectively. Thus malonyl-CoA resistance, which correlated strongly (0.678) with absolute oxidation rates (RG > SOL > EDL > WG), was greater in red than in white muscles. Similarly, malonyl-CoA resistant palmitate oxidation and CPT I activity were greater in mitochondria from RG compared with WG. Ribonuclease protection assays were performed to evaluate whether our data might be explained by differential expression of CPT I splice variants. We detected the presence of two CPT Ibeta splice variants that were more abundant in red compared with white muscle, but the relative expression of the two mRNA species was unrelated to malonyl-CoA resistance. These results provide evidence of a malonyl-CoA-insensitive CPT I activity in red muscle, suggesting fiber type-specific expression of distinct CPT I isoforms and/or posttranslational modulations that have yet to be elucidated. PMID- 11934666 TI - Contributions of total and regional fat mass to risk for cardiovascular disease in older women. AB - The aim of this study was to determine whether trunk fat mass, measured by dual energy X-ray absorptiometry (DEXA), is predictive of insulin resistance and dyslipidemia, independently of arm and leg fat mass, in postmenopausal women. Total and regional body composition was measured by DEXA in 166 healthy, postmenopausal women (66 +/- 4 yr). Four primary markers of insulin resistance and dyslipidemia were assessed: 1) area under the curve for the insulin (INS(AUC)) response to an oral glucose tolerance test (OGTT), 2) product of the OGTT glucose and insulin areas (INS(AUC)xGLU(AUC)), 3) serum triglycerides (TG), and 4) high-density lipoprotein (HDL)-cholesterol. Trunk fat mass was the strongest independent predictor of each of the primary dependent variables. In multivariate regression models, trunk fat mass was associated with unfavorable levels of INS(AUC), INS(AUC)xGLU(AUC), TG, and HDL-C, whereas leg fat mass was favorably associated with each of these variables. Thus trunk fat is a strong independent predictor of insulin resistance and dyslipidemia in postmenopausal women, whereas leg fat appears to confer protective effects against metabolic dysfunction. PMID- 11934667 TI - Aminoacyl-tRNA enrichment after a flood of labeled phenylalanine: insulin effect on muscle protein synthesis. AB - Muscle protein synthesis in dogs measured by flooding with L [(2)H(5)]phenylalanine (70 mg/kg) was significantly stimulated by infusion of insulin with amino acids. The stimulation of muscle protein synthesis was similar when calculated from the enrichment of phenylalanyl-tRNA (61 +/- 10%, P < 0.001), plasma phenylalanine (61 +/- 10%, P < 0.001), or tissue fluid phenylalanine (54 +/- 10%, P < 0.001). The time course for changes in enrichment of L [(2)H(5)]phenylalanine throughout the flooding period was determined for plasma, tissue fluid, and phenylalanyl-tRNA in the basal state and during the infusion of insulin with amino acids. Enrichments of plasma free phenylalanine and phenylalanyl-tRNA were equalized between 20 and 45 min, although the enrichment of phenylalanyl-tRNA was lower at early time points. Rates of muscle protein synthesis obtained with the flooding method and calculated from plasma phenylalanine enrichment were comparable to those calculated from phenylalanyl tRNA and also to those obtained previously with a continuous infusion of phenylalanine with phenylalanyl-tRNA as precursor. This study confirms that, with a bolus injection of labeled phenylalanine, the enrichment of aminoacyl-tRNA, the true precursor pool for protein synthesis, can be assessed from more readily sampled plasma phenylalanine. PMID- 11934668 TI - Accelerated rates of glycolysis in the hypertrophied heart: are they a methodological artifact? AB - Glycolysis, measured by (3)H(2)O production from [5-(3)H]glucose, is accelerated in isolated working hypertrophied rat hearts. However, nonglycolytic detritiation of [5-(3)H]glucose via the nonoxidative pentose phosphate pathway (PPP) could potentially lead to an overestimation of true glycolytic rates, especially in hypertrophied hearts where the PPP may be upregulated. To address this concern, we measured glycolysis using [5-(3)H]glucose and a second, independent method in isolated working hearts from halothane-anesthetized, sham-operated and aortic constricted rats. Glycolysis was accelerated in hypertrophied hearts compared with control hearts regardless of the method used. There was also excellent concordance in glycolytic rates between the different methods. Moreover, activity of glucose-6-phosphate dehydrogenase and expression of transaldolase, enzymes controlling key steps in the oxidative and nonoxidative PPP, respectively, were not different between control and hypertrophied hearts. Thus nonglycolytic detritiation of [5-(3)H]glucose in the PPP is insignificant, and (3)H(2)O production from [5-(3)H]glucose is an accurate means to measure glycolysis in isolated working normal and hypertrophied rat hearts. Furthermore, the PPP does not appear to be increased in cardiac hypertrophy induced by abdominal aortic constriction. PMID- 11934669 TI - Cr supplementation decreases tyrosine phosphorylation of the CreaT in skeletal muscle during sepsis. AB - Myocellular creatine (Cr) uptake is predominantly governed by a sodium-dependent Cr transporter (CreaT) and plays a pivotal role in skeletal muscle energy metabolism. The CreaT belongs to a neurotransmitter transporter family that can be functionally regulated by protein tyrosine kinase-induced tyrosine phosphorylation. The association between myocellular Cr and c-Src-related tyrosine phosphorylation of the CreaT and the influence of oral Cr supplementation on this association were investigated during sepsis. Animals were randomized to receive standard rat chow or standard rat chow with oral Cr supplementation for 4 days followed by cecal ligation and puncture (CLP) or sham operation. Fast-twitch gastrocnemius muscles were harvested 24 h after operation. Myocellular free Cr levels were 70% higher after CLP. Western blotting of the immunoprecipitated CreaT with an anti-phosphotyrosine or anti-phospho-c-Src (Y 416) antibody revealed that tyrosine phosphorylation of the CreaT and tyrosine phosphorylated c-Src (Tyr(416)) expression in the CreaT-c-Src complex were significantly increased after CLP compared with sham operation. These changes were observed in homogenates and plasma membrane fractions of gastrocnemius muscles. Although oral Cr supplementation increased myocellular free Cr levels equivalently in CLP and sham-operated animals, c-Src-related tyrosine phosphorylation of the CreaT in homogenates and plasma membrane fractions of gastrocnemius muscles was, however, downregulated in Cr-supplemented CLP animals compared with Cr-supplemented sham-operated rats. During sepsis, increased myocellular free Cr levels are associated with enhanced tyrosine phosphorylation of the CreaT, which is likely induced by active c-Src. Oral Cr supplementation downregulates c-Src-related tyrosine phosphorylation of the CreaT. The data suggest that myocellular Cr homeostasis and CreaT activity are tightly regulated and closely related during sepsis. PMID- 11934670 TI - Impact of high-fat diet and antioxidant supplement on mitochondrial functions and gene transcripts in rat muscle. AB - High-fat diets are reported to increase oxidative stress in a variety of tissues, whereas antioxidant supplementation prevents many diseases attributed to high-fat diet. Rodent skeletal muscle mitochondrial DNA has been shown to be a potential site of oxidative damage. We hypothesized that the effects of a high-fat diet on skeletal muscle DNA functions would be attenuated or partially reversed by antioxidant supplementation. Gene expression profiling and measurement of mitochondrial ATP production capacity were performed in skeletal muscle from male rats after feeding one of three diets (control, high-fat diet with or without antioxidants) for 36 wk. The high-fat diet altered transcript levels of 18 genes of 800 surveyed compared with the control-fed rats. Alterations included reduced expression of genes involved in free-radical scavenging and tissue development and increased expression of stress response and signal transduction genes. The magnitude of these alterations due to high-fat diet was reduced by antioxidant supplementation. Real-time PCR measurements confirmed the changes in transcript levels of cytochrome c oxidase subunit III and superoxide dismutase-1 and -2 noted by microarray approach. Mitochondrial ATP production was unaltered by dietary changes or antioxidant supplementation. It is concluded that the high-fat diet increases the transcription of genes involved in stress response but reduces those of free-radical scavenger enzymes, resulting in reduced DNA repair/metabolism (increased DNA damage). Antioxidants partially prevent these changes. Mitochondrial functions in skeletal muscle remain unaltered by the dietary intervention due to many adaptive changes in gene transcription. PMID- 11934671 TI - Muscle contraction increases lactate transport while reducing sarcolemmal MCT4, but not MCT1. AB - Rates of lactate uptake into giant sarcolemmal vesicles were determined in vesicles collected from rat muscles at rest and immediately after 10 min of intense muscle contraction. This contraction period reduced muscle glycogen rapidly by 37-82% in all muscles examined (P < 0.05) except the soleus muscle (no change P > 0.05). At an external lactate concentration of 1 mM lactate, uptake into giant sarcolemmal vesicles was not altered (P > 0.05), whereas at an external lactate concentration of 20 mM, the rate of lactate uptake was increased by 64% (P < 0.05). Concomitantly, the plasma membrane content of monocarboxylate transporter (MCT)1 was reduced slightly (-10%, P < 0.05), and the plasma membrane content of MCT4 was reduced further (-25%, P < 0.05). In additional studies, the 10-min contraction period increased the plasma membrane GLUT4 (P < 0.05) while again reducing MCT4 (-20%, P < 0.05) but not MCT1 (P > 0.05). These studies have shown that intense muscle contraction can increase the initial rates of lactate uptake, but only when the external lactate concentrations are high (20 mM). We speculate that muscle contraction increases the intrinsic activity of the plasma membrane MCTs, because the increase in lactate uptake occurred while plasma membrane MCT4 was decreased and plasma membrane MCT1 was reduced only minimally, or not at all. PMID- 11934672 TI - Hyposmotic shock stimulates insulin secretion by two distinct mechanisms. Studies with the betaHC9 cell. AB - Exposure of betaHC9 cells to a Krebs-Ringer bicarbonate-HEPES buffer (KRBH) made hypotonic by a reduction of 25 mM NaCl resulted in a prompt stimulation of insulin release. The stimulation was transient, and release rates returned to basal levels after 10 min. The response resembles that of the first phase of glucose-stimulated insulin release. The response did not occur if the reduction in NaCl was compensated for by the addition of an equivalent osmolar amount of sorbitol, so the stimulation of release was due to the osmolarity change and not the reduction in NaCl. The hyposmotic shock released insulin in KRBH with or without Ca(2+). The L-type Ca(2+) channel blocker nitrendipine inhibited the response in normal KRBH but had no effect in KRBH without Ca(2+) despite the latter response being larger than in the presence of extracellular Ca(2+). Similar data were obtained with calciseptine, which also blocks L-type channels. The T-type Ca(2+) channel blocker flunarizine was without effect, as was the chloride channel blocker DIDS. In parallel studies, the readily releasable pool of insulin-containing granules was monitored. Immunoprecipitation of the target SNARE protein syntaxin and co-immunoprecipitation of the vesicle-SNARE VAMP-2 was used as an indicator of the readily releasable granule pool. After hypotonic shock in the presence of extracellular Ca(2+), the amount of VAMP-2 coimmunoprecipitated by antibodies against syntaxin was much reduced compared with controls. Therefore, under these conditions, hypotonic shock stimulates exocytosis of the readily releasable pool of insulin-containing granules. No such reduction was seen in the absence of extracellular Ca(2+). In conclusion, after reexamination of the effect of hyposmotic shock on insulin secretion in the presence and absence of Ca(2+) (with EGTA in the medium), it is clear that two different mechanisms are operative under these conditions. Moreover, these two mechanisms may be associated with the release of two distinct pools of insulin containing granules. PMID- 11934673 TI - Individual variation in body temperature and energy expenditure in response to mild cold. AB - We studied interindividual variation in body temperature and energy expenditure, the relation between these two, and the effect of mild decrease in environmental temperature (16 vs. 22 degrees C) on both body temperature and energy expenditure. Nine males stayed three times for 60 h (2000-0800) in a respiration chamber, once at 22 degrees C and twice at 16 degrees C, in random order. Twenty four-hour energy expenditure, thermic effect of food, sleeping metabolic rate, activity-induced energy expenditure, and rectal and skin temperatures were measured. A rank correlation test with data of 6 test days showed significant interindividual variation in both rectal and skin temperatures and energy expenditures adjusted for body composition. Short-term exposure of the subjects to 16 degrees C caused a significant decrease in body temperature (both skin and core), an increase in temperature gradients, and an increase in energy expenditure. The change in body temperature gradients was negatively related to changes in energy expenditure. This shows that interindividual differences exist with respect to the relative contribution of metabolic and insulative adaptations to cold. PMID- 11934674 TI - Central leptin increases insulin sensitivity in streptozotocin-induced diabetic rats. AB - This study examined the effect of intracerebroventricular leptin on insulin sensitivity in streptozotocin (STZ)-induced diabetic rats. Male Wistar rats were cannulated in the lateral ventricle and, after recovery, administered either intravenous STZ (50 mg/kg) to induce diabetes or citrate buffer. Chronic leptin (10 microg/10 microl icv) or vehicle injections were administered daily for 14 days beginning 2 days after establishment of hyperglycemia in the diabetic animals. At the end of the 2 wk of injections, insulin sensitivity was measured by the steady-state plasma glucose (SSPG) method. Blood glucose concentrations were dramatically reduced and normalized by the 4th day in diabetic animals receiving intracerebroventricular leptin treatment. Diabetic animals exhibited insulin resistance, whereas intracerebroventricular leptin significantly enhanced insulin sensitivity, as indicated by decreased SSPG. Circulating leptin levels were not increased in animals injected with intracerebroventricular leptin. Thus the increased peripheral insulin sensitivity appears to be due solely to the presence of leptin in the brain, not to leptin acting peripherally. These data imply that inadequate central leptin signaling may lead to insulin resistance. PMID- 11934675 TI - Contribution of insulin to the translational control of protein synthesis in skeletal muscle by leucine. AB - Enhanced protein synthesis in skeletal muscle after ingestion of a balanced meal in postabsorptive rats is mimicked by oral leucine administration. To assess the contribution of insulin to the protein synthetic response to leucine, food deprived (18 h) male rats (approximately 200 g) were intravenously administered a primed-constant infusion of somatostatin (60 microg + 3 microg.kg(-1).h(-1)) or vehicle beginning 1 h before administration of leucine (1.35 g L-leucine/kg) or saline (control). Rats were killed 15, 30, 45, 60, or 120 min after leucine administration. Compared with controls, serum insulin concentrations were elevated between 15 and 45 min after leucine administration but returned to basal values by 60 min. Somatostatin maintained insulin concentrations at basal levels throughout the time course. Protein synthesis was increased between 30 and 60 min, and this effect was blocked by somatostatin. Enhanced assembly of the mRNA cap-binding complex (composed of eukaryotic initiation factors eIF4E and eIF4G) and hyperphosphorylation of the eIF4E-binding protein 1 (4E-BP1), the 70-kDa ribosomal protein S6 kinase (S6K1), and the ribosomal protein S6 (rp S6) were observed as early as 15 min and persisted for at least 60 min. Somatostatin attenuated the leucine-induced changes in 4E-BP1 and S6K1 phosphorylation and completely blocked the change in rp S6 phosphorylation but had no effect on eIF4G small middle dot eIF4E assembly. Overall, the results suggest that the leucine induced enhancement of protein synthesis and the phosphorylation states of 4E-BP1 and S6K1 are facilitated by the transient increase in serum insulin. In contrast, assembly of the mRNA cap-binding complex occurs independently of increases in insulin and, by itself, is insufficient to stimulate rates of protein synthesis in skeletal muscle after leucine administration. PMID- 11934676 TI - Impaired fatty acid oxidation in muscle of aging rats perfused under basal conditions. AB - The purpose of the present study was to examine the utilization of fatty acids (FA) and muscle substrates by skeletal muscle in young, middle-aged, and old adult rats under conditions of euglycemia with low insulin levels. Male Fischer 344 x Brown Norway rats aged 5, 15, or 24 mo underwent hindlimb perfusion with a medium of 8 mM glucose, 1 mM palmitate, 25 microU/ml insulin, [1-(14)C]palmitate, and [3-(3)H]glucose. Glucose and palmitate uptake were similar among age groups. The percent and total palmitate oxidized (nmol.min(-1).g(-1)) were 30-36 and 41 49% lower (P < 0.05) in 15-mo- and 24-mo-old than in 5-mo-old animals. Compared with 5-mo- and 15-mo-old animals, pre- and postperfusion muscle triglyceride (TG) levels were significantly (P < 0.05) elevated 91-305% in red and 118-219% in white muscles of 24-mo-old animals. Fatty acid-binding protein content was 40-64% higher (P < 0.05) in 24-mo- than in 5-mo- or 15-mo-old animals. In red muscle, hormone-sensitive lipase (HSL) content was 28% lower (P < 0.05) in 24-mo- than in 5-mo-old animals. These results indicate that, under euglycemic conditions in the presence of low insulin levels, the reduction in FA disposal to oxidation and the decrease in HSL content may contribute to the accumulation of TG in muscle of old animals. PMID- 11934677 TI - Opposing effects of pituitary leukemia inhibitory factor and SOCS-3 on the ACTH axis response to inflammation. AB - We have shown that leukemia inhibitory factor (LIF) and suppressor of cytokine signaling (SOCS)-3 are expressed in the hypothalamus and pituitary and that LIF induces proopiomelanocortin (POMC) and ACTH, whereas SOCS-3 abrogates corticotroph POMC gene transcription and ACTH secretion. Here, we determined the role of pituitary LIF and SOCS-3 in regulating hypothalamo-pituitary-adrenal (HPA) axis inflammatory responses. Murine pituitary LIF expression was induced up to eightfold after intraperitoneal injection of lipopolysaccharide or tumor necrosis factor-alpha, concordant with elevated plasma levels of ACTH and corticosterone. In LIF knockout (LIFKO) mice, induction of both ACTH and corticosterone were attenuated. LIF deletion was associated with elevated (P < 0.05) levels of pituitary TNF-alpha, interleukin (IL)-1beta, and IL-6 mRNA and cytokine-inducible pituitary SOCS-3 expression. Abrogation of the HPA axis stress response and higher pituitary levels of proinflammatory cytokines observed in LIFKO mice resulted in a stronger inflammatory process, as evidenced by elevated erythrocyte sedimentation rate and increased serum amyloid A levels (P < 0.05). The results indicate that, although LIF induces ACTH, SOCS-3 acts to counterregulate the HPA axis response to inflammation. PMID- 11934678 TI - Triiodothyronine is required for the stimulation of type II 5'-deiodinase mRNA in rat brown adipocytes. AB - Type II 5'-iodothyronine deiodinase (D2), produces triiodothyronine (T(3)) and is stimulated by cold exposure via norepinephrine (NE) release in brown adipose tissue. Cultured rat brown adipocytes require T(3) for the adrenergic stimulation of D2 activity. D2 mRNA expression in cultured brown adipocytes is undetectable with the use of basal conditions or NE without T(3). Full D2 expression is achieved using NE + T(3), especially after prolonged T(3) exposure. beta(3) Adrenergic agonists mimic the NE action, whereas cAMP analogs do not. Prolonged exposure to T(3) alone increases D2 mRNA. High T(3) doses (500 nM) inhibit the adrenergic stimulation of D2 activity while increasing D2 mRNA. The effects obtained with NE + T(3) or T(3) alone are suppressed by actinomycin, but not by cycloheximide, which leads to accumulation of short D2 mRNA transcripts. Prolonged or short exposure to T(3) did not change D2 mRNA half-life, but T(3) seemed to elongate it. In conclusion, T(3) is an absolute requirement for the adrenergic stimulation of D2 mRNA in brown adipocytes. T(3) upregulates D2 mRNA, an effect that might involve stimulation of factors required for transcription or for stabilization of D2 mRNA. PMID- 11934679 TI - Prior exercise and the response to insulin-induced hypoglycemia in the dog. AB - To test whether hepatic insulin action and the response to an insulin-induced decrement in blood glucose are enhanced in the immediate postexercise state as they are during exercise, dogs had sampling (artery, portal vein, and hepatic vein) catheters and flow probes (portal vein and hepatic artery) implanted 16 days before a study. After 150 min of moderate treadmill exercise or rest, dogs were studied during a 150-min hyperinsulinemic (1 mU.kg(-1).min(-1)) euglycemic (n = 5 exercised and n = 9 sedentary) or hypoglycemic (65 mg/dl; n = 8 exercised and n = 9 sedentary) clamp. Net hepatic glucose output (NHGO) and endogenous glucose appearance (R(a)) and utilization (R(d)) were assessed with arteriovenous and isotopic ([3-(3)H]glucose) methods. Results show that, immediately after prolonged, moderate exercise, in relation to sedentary controls: 1) the glucose infusion rate required to maintain euglycemia, but not hypoglycemia, was higher; 2) R(d) was greater under euglycemic, but not hypoglycemic conditions; 3) NHGO, but not R(a), was suppressed more by a hyperinsulinemic euglycemic clamp, suggesting that hepatic glucose uptake was increased; 4) a decrement in glucose completely reversed the enhanced suppression of NHGO by insulin that followed exercise; and 5) arterial glucagon and cortisol were transiently higher in the presence of a decrement in glucose. In summary, an increase in insulin action that was readily evident under euglycemic conditions after exercise was abolished by moderate hypoglycemia. The means by which the glucoregulatory system is able to overcome the increase in insulin action during moderate hypoglycemia is related not to an increase in R(a) but to a reduction in insulin-stimulated R(d). The primary site of this reduction is the liver. PMID- 11934681 TI - Immediate effects of an 8-h advance shift of the rest-activity cycle on 24-h profiles of cortisol. AB - To investigate the adaptation of plasma cortisol profiles to an abrupt phase advance of the rest-activity cycle, eight normal young subjects were submitted in a sleep laboratory to an 8-h advance shift of their sleep-wake and dark-light cycles. The shift was achieved by advancing bedtimes from 2300-0700 to 1500-2300. Blood samples were obtained at 20-min intervals for 68 consecutive hours. The shift resulted within 6-9 h in a 3- to 4-h advance of timings of the nadir of the cortisol profile and of the end of the quiescent period but had no immediate effect on the timing of cortisol acrophase. The quiescent period of cortisol secretion was shortened and fragmented. Thus a major advance shift achieved without enforcing sleep deprivation results in a rapid partial adaptation of the temporal profiles of cortisol but also in a marked disruption of the cortisol quiescent period. Sleep onset was consistently followed by a decrease in cortisol concentrations. Conversely, both sleep-wake and dark-light transitions were consistently associated with cortisol secretory pulses. PMID- 11934680 TI - Effect of the ovarian hormones on GLUT4 expression and contraction-stimulated glucose uptake. AB - This study examined the roles of the female sex steroids, 17beta-estradiol (E(2)) and progesterone (Prog), on glucose uptake and GLUT4 protein expression. Female Sprague-Dawley rats were either sham operated (C) or ovariectomized and treated with placebo (O), E(2) (E), Prog (P), or both hormones at physiological doses (P + E) or the same dose of Prog with a high dose of E(2) (P + HiE) via timed release pellets inserted at the time of surgery, 15 days before metabolic testing. On the morning of day 15, animals received a 300-microCi injection (ip) of 2-deoxy-[(14)C]glucose and then either exercised on a motorized treadmill for 30 min at 0.35 m/s or remained sedentary in their cages for the same period. Basal glucose uptake was not different between the treatment groups in either the red or white quadriceps. However, glucose uptake was decreased (P < 0.05) in O, P, and P + E rats during exercise in the red quadriceps compared with C rats, whereas E and P + HiE treatment restored glucose uptake. Glycogen content in skeletal muscle followed similar trends, with no differences seen in resting animals. Postexercise red quadriceps glycogen levels were higher (P < 0.05) in the E and P + HiE rats compared with O and P. Treatment of ovariectomized rats with progesterone (P rats) decreased (P < 0.05) GLUT4 content in the red quadriceps by 21% compared with C rats. These data demonstrate that estrogen deficient animals have a decreased ability for contraction-stimulated glucose uptake and increased glycogen use during aerobic exercise. However, changes in contraction-stimulated glucose uptake could not be explained by altered transporter protein content, since the absence of E(2) had no effect on GLUT4 protein. PMID- 11934683 TI - Regional myocardial blood flow and glucose utilization during fasting and physiological hyperinsulinemia in humans. AB - We investigated the effect of insulin on total and regional myocardial blood flow (MBF) and glucose uptake (MGU) in healthy subjects (50 +/- 5 yr) by means of positron emission tomography (PET) with oxygen-15-labeled water (H(2)(15)O) and fluorine-18 labeled fluorodeoxyglucose ((18)FDG) before and during physiological hyperinsulinemia (40 mU.min(-1).m(-2)). Twelve male subjects were included in the study. During hyperinsulinemia, MBF increased from 0.91 +/- 0.28 to 1.01 +/- 0.31 ml.min(-1).g(-1) (n = 7 patients, P = 0.05; n = 112 regions, P < 0.005). Intersubject variability ranged from -3.0 to +41%. MGU increased from 0.11 +/- 0.08 (n = 5) to 0.56 +/- 0.08 micromol.min(-1).g(-1) (P < 0.0001, n = 7). MBF and insulin-mediated MGU were higher in the septum and anterior and lateral wall along short-axis regions of the heart. During hyperinsulinemia, MBF was also higher in the apex and midventricle compared with the base. MBF and MGU were positively correlated before (r = 0.66, P < 0.0001) and during hyperinsulinemia (r = 0.24, P < 0.05). These results provide evidence that insulin stimulates MBF in normal human hearts and appears to involve mainly those regions of the heart where insulin-mediated MGU is higher. Furthermore, regional distribution of insulin-stimulated MBF and MGU does not appear to be uniform across the left ventricular wall of healthy subjects. PMID- 11934682 TI - Effect of IGF-I therapy on VLDL apolipoprotein B100 metabolism in type 1 diabetes mellitus. AB - Abnormal lipid metabolism may be related to the increased cardiovascular risk in type 1 diabetes. Secretion and clearance rates of very low density lipoprotein (VLDL) apolipoprotein B100 (apoB) determine plasma lipid concentrations. Type 1 diabetes is characterized by increased growth hormone (GH) secretion and decreased insulin-like growth factor (IGF) I concentrations. High-dose IGF-I therapy improves the lipid profile in type 1 diabetes. This study examined the effect of low-dose (40 microg.kg(-1).day(-1)) IGF-I therapy on VLDL apoB metabolism, VLDL composition, and the GH-IGF-I axis during euglycemia in type 1 diabetes. Using a stable isotope technique, VLDL apoB kinetics were estimated before and after 1 wk of IGF-I therapy in 12 patients with type 1 diabetes in a double-blind, placebo-controlled trial. Fasting plasma triglyceride (P < 0.03), VLDL-triglyceride concentrations (P < 0.05), and the VLDL-triglyceride-to-VLDL apoB ratio (P < 0.002) significantly decreased after IGF-I therapy, whereas VLDL apoB kinetics were not significantly affected by IGF-I therapy. IGF-I therapy resulted in a significant increase in IGF-I and a significant reduction in GH concentrations. The mean overnight insulin concentrations during euglycemia decreased by 25% after IGF-I therapy. These results indicate that low-dose IGF-I therapy restores the GH-IGF-I axis in type 1 diabetes. IGF-I therapy changes fasting triglyceride concentrations and VLDL composition probably because of an increase in insulin sensitivity. PMID- 11934684 TI - Peptide transport in the mammary gland: expression and distribution of PEPT2 mRNA and protein. AB - The lactating mammary gland utilizes free plasma amino acids as well as those derived by hydrolysis from circulating short-chain peptides for protein synthesis. Apart from the major route of amino acid nitrogen delivery to the gland by the various transporters for free amino acids, it has been suggested that dipeptides may also be taken up in intact form to serve as a source of amino acids. The identification of peptide transporters in the mammary gland may therefore provide new insights into protein metabolism and secretion by the gland. The expression and distribution of the high-affinity type proton-coupled peptide transporter PEPT2 were investigated in rat lactating mammary gland as well as in human epithelial cells derived from breast milk. By use of RT-PCR, PEPT2 mRNA was detected in rat mammary gland extracts and human milk epithelial cells. The expression pattern of PEPT2 mRNA revealed a localization in epithelial cells of ducts and glands by nonisotopic high resolution in situ hybridization. In addition, immunohistochemistry was carried out and showed transporter immunoreactivity in the same epithelial cells of the glands and ducts. In addition, two-electrode voltage clamp recordings using PEPT2-expressing Xenopus laevis oocytes demonstrated positive inward currents induced by selected dipeptides that may play a role in aminonitrogen handling in mammalian mammary gland. Taken together, these data suggest that PEPT2 is expressed in mammary gland epithelia, in which it may contribute to the reuptake of short-chain peptides derived from hydrolysis of milk proteins secreted into the lumen. Whereas PEPT2 also transports a variety of drugs, such as selected beta-lactams, angiotensin-converting enzyme inhibitors, and antiviral and anticancer metabolites, their efficient reabsorption via PEPT2 may reduce the burden of xenobiotics in milk. PMID- 11934685 TI - Amelioration of high fructose-induced metabolic derangements by activation of PPARalpha. AB - To elucidate molecular mechanisms of high fructose-induced metabolic derangements and the influence of peroxisome proliferator-activated receptor-alpha (PPARalpha) activation on them, we examined the expression of sterol regulatory element binding protein-1 (SREBP-1) and PPARalpha as well as its nuclear activation and target gene expressions in the liver of high fructose-fed rats with or without treatment of fenofibrate. After 8-wk feeding of a diet high in fructose, the mRNA contents of PPARalpha protein and its activity and gene expressions of fatty acid oxidation enzymes were reduced. In contrast, the gene expressions of SREBP-1 and lipogenic enzymes in the liver were increased by high fructose feeding. Similar high fructose effects were also found in isolated hepatocytes exposed to 20 mM fructose in the media. The treatment of fenofibrate (30 mg.kg(-1).day(-1)) significantly improved high fructose-induced metabolic derangements such as insulin resistance, hypertension, hyperlipidemia, and fat accumulation in the liver. Consistently, the decreased PPARalpha protein content, its activity, and its target gene expressions found in high fructose-fed rats were all improved by fenofibrate treatment. Furthermore, we also found that the copy number of mitochondrial DNA, the expressions of mitochondrial transcription factor A, ATPase-6 subunit, and uncoupling protein-3 were increased by fenofibrate treatment. These findings suggest that the metabolic syndrome in high fructose fed rats is reversed by fenofibrate treatment, which is associated with the induction of enzyme expression related to beta-oxidation and the enhancement of mitochondrial gene expression. PMID- 11934686 TI - Role of nitric oxide in the regulation of nephron transport. AB - Nitric oxide (NO) plays an important role in various physiological processes in the kidney. In vivo experiments first suggested that the natriuretic and diuretic effects caused by NO may be due to decreased NaCl and fluid absorption by the nephron. In the last 10 years, several reports have directly demonstrated a role for NO in modulating transport in different tubule segments. The effects of NO on proximal tubule transport are still controversial. Both stimulation and inhibition of net fluid and bicarbonate have been reported in this segment, whereas only inhibitory effects of NO have been found in Na/H exchanger and Na/K ATPase activity. The effects of NO in the thick ascending limb are more homogeneous than in the proximal tubule. In this segment, NO decreases net Cl and bicarbonate absorption. A direct inhibitory effect of NO on the Na-K-2Cl cotransporter and the Na/H exchanger has been reported, while NO was found to stimulate apical K channels in this segment. In the collecting duct, NO inhibits Na absorption and vasopressin-stimulated osmotic water permeability. An inhibitory effect of NO on H-ATPase has also been reported in intercalated cells of the collecting duct. Overall, the reported effects of NO in the different nephron segments mostly agree with the natriuretic and diuretic effects observed in vivo. However, the net effect of NO on transport is still controversial in some segments, and in cases like the distal tubule, it has not been studied. PMID- 11934687 TI - Expression of myosin VI within the early endocytic pathway in adult and developing proximal tubules. AB - Myosin VI is a reverse-direction molecular motor implicated in membrane transport events. Because myosin VI is most highly expressed in the kidney, we investigated its renal localization by using high-resolution immunocytochemical and biochemical methods. Indirect immunofluorescence microscopy revealed myosin VI at the base of the brush border in proximal tubule cells. Horseradish peroxidase uptake studies, which labeled endosomes, and double staining for clathrin adapter protein-2 showed that myosin VI was closely associated with the intermicrovillar (IMV) coated-pit region of the brush border. Localization of myosin VI to the IMV region was confirmed at the electron microscopic level by colloidal gold labeling of ultrathin cryosections. In addition, antigen retrieval demonstrated a small but significant pool of myosin VI on the microvilli. To confirm the association of myosin VI with the IMV compartment, these membranes were separated from other membrane compartments by using 15-25% OptiPrep density gradients. Immunoblotting of the gradient fractions confirmed that myosin VI was enriched with markers for the IMV microdomain of the brush border, suggesting that myosin VI associates with proteins in this compartment. Finally, we examined the expression of myosin VI during nephron development. We found myosin VI present in a diffuse cytoplasmic pattern at stage II (S-shaped body phase) and that it was only redistributed fully to the brush border in the stage IV nephron. These studies support a model for myosin VI function in the endocytic process of the proximal tubule. PMID- 11934688 TI - alpha-Melanocyte-simulating hormone and interleukin-10 do not protect the kidney against mercuric chloride-induced injury. AB - The anti-inflammatory cytokines alpha-melanocyte-stimulating hormone (MSH) and interleukin (IL)-10 inhibit acute renal failure (ARF) after ischemia or cisplatin administration; however, these agents have not been tested in a pure nephrotoxic model of ARF. Therefore, we examined the effects of alpha-MSH and IL-10 in HgCl(2)-induced ARF. Mice were injected subcutaneously with HgCl(2) and then given vehicle, alpha-MSH, or IL-10 by intravenous injection. Animals were killed to study serum creatinine, histology, and myeloperoxidase activity. Treatment with either alpha-MSH or IL-10 did not alter the increase in serum creatinine, tubular damage, or leukocyte accumulation at 48 h after HgCl(2) injection. Because alpha-MSH and IL-10 are active in other injury models that involve leukocytes, we studied the time course of tubular damage and leukocyte accumulation to investigate whether leukocytes caused the tubular damage or accumulated in response to the tubular damage. Tubular damage was present in the outer stripe 12 h after HgCl(2) injection. In contrast, the number of leukocytes and renal myleoperoxidase activity were normal at 12 h but were significantly increased at 24 and 48 h after injection. We conclude that neither alpha-MSH nor IL-10 altered the course of HgCl(2)-induced renal injury. Because the tubular damage preceded leukocyte infiltration, the delayed leukocyte accumulation may play a role in the removal of necrotic tissue and/or tissue repair in HgCl(2) induced ARF. PMID- 11934689 TI - Mouse TonEBP-NFAT5: expression in early development and alternative splicing. AB - Tonicity-responsive enhancer binding protein (TonEBP)- nuclear factor of activated T cell family 5 is a DNA binding protein that plays a key role in the response of cells to hypertonicity. However, TonEBP is expressed and active in tissues that are in an isotonic milieu. To explore the biological role of TonEBP, we cloned mouse TonEBP that shares 92% of amino acids with the human counterpart. TonEBP is expressed in embryonic stem cells and throughout the stages of fetal development. Immunohistochemical analysis shows expression of TonEBP in most, if not all, developing tissues, including the brain, colon, heart, muscle, and eyes. Widespread alternative splicing in exons 2-4 was detected throughout development and in different adult tissues. As a result, four different polypeptides are produced with different lengths at the NH(2) terminus. Two of the isoforms differ in their ability to stimulate transcription. In conclusion, the presence of TonEBP mRNA during mouse embryogenesis suggests that TonEBP functions at all stages of mouse development, as well as in isotonic adult tissues. PMID- 11934690 TI - Impaired trafficking of distal renal tubular acidosis mutants of the human kidney anion exchanger kAE1. AB - Distal renal tubular acidosis (dRTA) is an inherited disease characterized by the failure of the kidneys to appropriately acidify urine and is associated with mutations in the anion exchanger (AE)1 gene. The effect of the R589H dRTA mutation on the expression of the human erythroid AE1 and the truncated kidney form (kAE1) was examined in transfected human embryonic kidney 293 cells. AE1, AE1 R589H, and kAE1 were present at the cell surface, whereas kAE1 R589H was located primarily intracellularly as shown by immunofluorescence, cell surface biotinylation, N-glycosylation, and anion transport assays. Coexpression of kAE1 R589H reduced the cell surface expression of kAE1 and AE1 by a dominant-negative effect, due to heterodimer formation. The mutant AE1 and kAE1 bound to an inhibitor affinity resin, suggesting that they were not grossly misfolded. Other mutations at R589 also prevented the formation of the cell surface form of kAE1, indicating that this conserved arginine residue is important for proper trafficking. The R589H dRTA mutation creates a severe trafficking defect in kAE1 but not in erythroid AE1. PMID- 11934691 TI - Renal interstitial hydrostatic pressure and natriuretic responses to volume expansion in pregnant rats. AB - During normal pregnancy, a gradual plasma volume expansion (VE) occurs and reaches a maximum level at late term. Pressure natriuresis and renal interstitial hydrostatic pressure (RIHP) responses are attenuated in pregnant rats. Also, basal RIHP is lower in pregnant rats, suggesting an increase in renal interstitial compliance during pregnancy. This adaptation may contribute to the increase in plasma volume that is required for a normal pregnancy, because increases in RIHP have been consistently shown to produce natriuresis and diuresis. Acute saline VE (5% body wt/30 min) has been shown to increase RIHP in normal nonpregnant rats. Therefore, the objective of this study was to determine RIHP, natriuretic, and diuretic responses to VE in nonpregnant (n = 7), midterm pregnant (n = 8), and late-term pregnant (n = 8) Sprague-Dawley rats. Although VE significantly increased RIHP, fractional excretion of sodium (FE(Na)), and urine flow rate (V) in all groups, DeltaRIHP was highest for nonpregnant (3.0 +/- 0.3 mmHg) compared with midterm pregnant (1.6 +/- 0.1 mmHg; P < 0.05 vs. nonpregnant) and late-term pregnant rats (1.2 +/- 0.1 mmHg; P < 0.05 vs. both midterm pregnant and nonpregnant rats). DeltaFE(Na) and DeltaV were similar in all groups: 5.8 +/- 1.0% and 231 +/- 27 microl/min for nonpregnant, 6.8 +/- 1.3% and 173 +/- 16 microl/min for midterm pregnant, and 7.6 +/- 1.2% and 203 +/- 10 microl/min for late-term pregnant rats, respectively. In conclusion, basal RIHP and the increase in RIHP during VE were attenuated during pregnancy; however, the natriuretic and diuretic responses to VE remain intact during the course of pregnancy. PMID- 11934692 TI - Structural determinants and specificities for ROMK1-phosphoinositide interaction. AB - We have recently reported that direct interaction between phosphatidylinositol bisphosphate (PIP(2)) and the COOH-terminal cytoplasmic domain of ROMK1 is important for opening of the channel. We identified arginine-188 of ROMK1 as a critical residue for this interaction. Here, we further report that substitution of a neutral amino acid for lysine-181, arginine-217, or lysine-218 decreases single-channel open probability for the full-conductance state and increases the frequency of opening at a subconductance state. Compared with wild-type ROMK1 channels, these substitution mutants also display an increased sensitivity to the block by anti-PIP(2) antibodies and to inhibition by intracellular protons. These results indicate that, like arginine-188, lysine-181, arginine-217, and lysine 218 are also involved in interactions with PIP(2) and are critical for ROMK1 to open at full conductance. Using synthetic phosphoinositides containing phosphates at different positions in the head group, we also examined the specificities of phosphoinositides in the regulation of ROMK1 channels. We found that phosphoinositides containing phosphate at both positions 4 and 5 of the inositol head group have the highest efficacy in activating ROMK1 channels. These results suggest that phosphatidylinositol 4,5-bisphosphate is likely the important phosphoinositide in the regulation of ROMK1 channels in a physiological membrane milieu. PMID- 11934693 TI - Rat proximal NHE3 adapts to chronic acid-base disorders but not to chronic changes in dietary NaCl intake. AB - In the proximal tubule, the apical Na(+)/H(+) exchanger identified as NHE3 mediates most NaCl and NaHCO(3) absorption. The purpose of this study was to analyze the long-term regulation of NHE3 during alkalosis induced by dietary NaHCO(3) loading and changes in NaCl intake. Sprague-Dawley rats exposed to a low NaCl, high-NaCl, or NaHCO(3) diet for 6 days were studied. Renal cortical apical membrane vesicles (AMV) were prepared from treated and normal rats. Na(+)/H(+) exchange was assayed as the initial rate of (22)Na(+) uptake in the presence of an outward H(+) gradient. (22)Na(+) uptake measured in the presence of high-dose 5-(N-ethyl-N-isopropyl) amiloride was not different among models. Changes in NaCl intake did not affect NHE3 activity, whereas NaHCO(3) loading inhibited (22)Na(+) uptake by 30%. AMV NHE3 protein abundance assessed by Western blot analysis was unaffected during changes in NaCl intake. During NaHCO(3) loading, NHE3 protein abundance was decreased by 65%. We conclude that proximal NHE3 adapts to chronic metabolic acid-base disorders but not to changes in dietary NaCl intake. PMID- 11934694 TI - Adenosine attenuates oxidant injury in human proximal tubular cells via A(1) and A(2a) adenosine receptors. AB - We have recently demonstrated protection against renal ischemic-reperfusion injury in vivo by A(1)- and A(2a)-adenosine receptor (AR) modulations. To further elucidate the signaling cascades of AR-induced cytoprotection against reperfusion/oxidant-mediated injury, immortalized human proximal tubule (HK-2) cells were treated with H(2)O(2). H(2)O(2) caused dose- and time-dependent HK-2 cell death that was measured by lactate dehydrogenase release and trypan blue dye uptake. Adenosine protected against H(2)O(2)-induced HK-2 cell death by means of A(1)- and A(2a)-AR activation. A(1)-AR-mediated protection involves pertussis toxin-sensitive G proteins and protein kinase C, whereas A(2a)-AR-mediated protection involves protein kinase A activation by means of cAMP and activation of the cAMP response element binding protein. Moreover, protein kinase A activators (forskolin and Sp-isomer cAMP) also protected HK-2 cells against H(2)O(2) injury. De novo gene transcription and protein synthesis are required for both A(1)- and A(2a)-AR-mediated cytoprotection as actinomycin D and cycloheximide, respectively, blocked cytoprotection. Chronic treatments with a nonselective AR agonist abolished the protection by adenosine. Moreover, chronic treatments with a nonselective AR antagonist increased the endogenous tolerance of HK-2 cells against H(2)O(2). We concluded that A(1)- and A(2a)-AR activation protects HK-2 cells against H(2)O(2)-induced injury by means of distinct signaling pathways that require new gene transcription and new protein synthesis. PMID- 11934695 TI - Glucocorticoids inhibit transcription and expression of the UT-A urea transporter gene. AB - Dexamethasone treatment increases urea excretion and decreases urea permeability and urea transporter UT-A1 protein abundance in the inner medullary collecting duct (IMCD) of adrenalectomized rats. We examined the effect of dexamethasone treatment for 3 days on the abundance of several UT-A mRNA transcripts in rat renal medulla. By Northern blot analysis, a significant decrease in mRNA expression was observed in the inner medulla of dexamethasone-treated rats compared with controls for UT-A1 (71%), UT-A3 (75%), and UT-A3b (75%), but not for UT-A2. We then tested the effect of 100 nM dexamethasone on the activity of promoter I in the UT-A gene, using LLC-PK(1)-GR101 cells that express the glucocorticoid receptor. Dexamethasone significantly decreased the activity of rat UT-A promoter I (72%) but did not affect UT-A promoter II. Deletion analysis and site-directed mutagenesis demonstrated that sequences between -423 and -244 are important for this inhibition and that a 10-bp sequence at -363, which binds a nuclear protein in a gel shift assay, is necessary for basal promoter activity. The specific factors involved in repression of UT-A promoter I activity by glucocorticoids remain to be determined. PMID- 11934696 TI - Stenosis-dependent role of nitric oxide and prostaglandins in chronic renal ischemia. AB - The role of nitric oxide (NO) and prostaglandins (PG) in modifying renal hemodynamics was examined in clipped and nonclipped kidneys of unilateral renal artery stenosis. Chronic unilateral renal ischemia was established by 4-wk clipping the left renal artery of canine kidneys, and renal interstitial nitrate+nitrite and PGE(2) contents were evaluated by the microdialysis technique. Unilateral renal artery stenosis caused 45 +/- 1 and 73 +/- 1% decrements in renal plasma flow (RPF) in moderately and severely clipped kidneys and 21 +/- 3% decrements in nonclipped kidneys with severe stenosis. Renal nitrate+nitrite decreased in moderately (-31 +/- 1%) and severely clipped kidneys (-63 +/- 4%). N(omega)-nitro-L-arginine methyl ester reduced RPF (-56 +/- 3%) and glomerular filtration rate (GFR; -54 +/- 3%) in moderately clipped kidneys, whereas this inhibitory effect was abolished in severely clipped kidneys. In contrast, renal PGE(2) contents increased modestly in moderate clipping and were markedly elevated in severely clipped kidneys (from 111 +/- 7 to 377 +/- 22 pg/ml); sulpyrine impaired renal hemodynamics only in severely clipped kidneys. In contralateral nonclipped kidneys, although renal PGE(2) was not increased, sulpyrine reduced RPF (-32 +/- 1%) and GFR (-33 +/- 3%) in severe stenosis. Collectively, NO plays a substantial role in maintaining renal hemodynamics both under basal condition and in moderate renal artery stenosis, whereas the contributory role shifts from NO to PG as renal artery stenosis progresses. Furthermore, because intrarenal angiotensin II is reported to increase in nonclipped kidneys, unilateral severe ischemia may render the nonclipped kidney susceptible to PG inhibition. PMID- 11934697 TI - Properties of an inwardly rectifying K(+) channel in the basolateral membrane of mouse TAL. AB - We investigated the properties of K(+) channels in the basolateral membrane of the cortical thick ascending limb (CTAL) using the patch-clamp technique. Approximately 34% of cell-attached patches contained an inwardly rectifying K(+) channel (K(+)-to-Na(+) permeability ratio approximately 22), having an inward conductance (G(in)) of 44 pS and an outward conductance (G(out)) of approximately 10 pS (G(in)/G(out) approximately 4). Channel activity (NP(o)) increased with depolarization. When the cytosolic sides of inside-out patches were exposed to an Mg(2+)-free medium, the channel had a G(in) of 50 pS and was weakly inwardly rectifying (G(in)/G(out) approximately 1). Cytosolic Mg(2+) reduced G(out), yielding a G(in)/G(out) of 3.8 at 1.3 mM Mg(2+). Internal Na(+) also yielded a G(in)/G(out) of 1.6 at 20 mM Na(+). Spermine reduced NP(o) on inside-out membrane patches. Sensitivity to spermine at depolarizing voltages [half-maximal inhibitory concentration (K(i)) = 0.2 microM] was much greater than at hyperpolarizing voltages (K(i) = 26 microM). Half-inactivation by 0.5 microM spermine occurred at a clamp potential of 43 mV, with an effective valence of 1.25. A sigmoid relationship between bath pH and NP(o) of inside-out membrane patches was observed, with a pK of 7.6 and a Hill coefficient of 1.8. Intracellular acidification also reduced the NP(o) of cell-attached patches. This channel is probably a major component of K(+) conductance in the CTAL basolateral membrane. PMID- 11934698 TI - Confocal imaging of organic anion transport in intact rat choroid plexus. AB - We used confocal microscopy and quantitative image analysis to follow the movement of the fluorescent organic anion fluorescein (FL) from bath to cell and cell to blood vessel in intact rat lateral choroid plexus. FL accumulation in epithelial cells and underlying vessels was rapid, concentrative, and reduced by other organic anions. At steady state, cell fluorescence exceeded bath fluorescence by a factor of 3-5, and vessel fluorescence exceeded cell fluorescence by a factor of approximately 2. In cells, FL distributed between diffuse and punctate compartments. Cell and vessel accumulation of FL decreased when metabolism was inhibited by KCN, when bath Na(+) was reduced from 130 to 26 mM, and when the Na(+) gradient was collapsed with ouabain. Cell and vessel accumulation increased by >50% when 1-10 microM glutarate was added to the bath. Finally, transport of FL and carboxyfluorescein (generated intracellularly from carboxyfluorescein diacetate) from cell to blood vessel was greatly diminished when medium K(+) concentration ([K(+)]) was increased 10-fold. These results 1) validate a new approach to the study of choroid plexus function, and 2) indicate a two-step mechanism for transepithelial organic anion transport: indirect coupling of uptake to Na(+) at the apical membrane and electrical potential driven efflux at the basolateral membrane. PMID- 11934699 TI - Age dependency of renal function in CD-1 mice. AB - Renal function was studied in mice of different ages. In metabolic cage experiments, the renal electrolyte excretion was similar in young (n = 8; 5- to 7 wk-old) and adult (n = 6; 20- to 22-wk-old) CD-1 (ICR) BR mice, whereas spontaneous drinking volume and urinary flow rate were significantly higher in the adult compared with the young mice. Subsequently, the renal functional reserve was investigated by amino acid (AA) infusion (10%) in anesthetized young (n = 8) and adult (n = 6) mice. Because the body weight of adult mice was significantly higher than that of young animals, one group of adult mice (n = 8) received 12.5% AA to ensure that the dose of AA related to body weight was similar in both groups. Young animals constantly infused with Ringer solution served as time controls (n = 8). Glomerular filtration rate (GFR) at baseline was similar in each group. Because of AA, GFR significantly increased in young mice but not in both groups of adult animals, whereas in time controls GFR remained constant. Urinary flow rate and sodium excretion were elevated by AA in young and adult mice. We conclude that in CD-1 mice the first signs of age-related changes in kidney function concern alterations in renal hemodynamics, whereas renal tubular function appears to be preserved. PMID- 11934701 TI - Genomic organization of the 5' end of human beta-ENaC and preliminary characterization of its promoter. AB - The mRNA for the beta-subunit of the epithelial Na(+) channel (beta-ENaC) is regulated developmentally and, in some tissues, in response to corticosteroids. To understand the mechanisms of transcriptional regulation of the human beta-ENaC gene, we characterized the 5' end of the gene and its 5'-flanking regions. Adaptor-ligated human kidney and lung cDNA were amplified by 5' rapid amplification of cDNA ends, and transcription start sites of two 5' variant transcripts were determined by nuclease protection or primer extension assays. Cosmid clones that contain the 5' end of the gene were isolated, and analysis of these clones indicated that alternate first exons approximately 1.5 kb apart and approximately 45 kb upstream of a common second exon formed the basis of these transcripts. Genomic fragments that included the proximal 5'-flanking region of either transcript were able to direct expression of a reporter gene in lung epithelia and to bind Sp1 in nuclear extracts, confirming the presence of separate promoters that regulate beta-ENaC expression. PMID- 11934700 TI - PAR-2 elicits afferent arteriolar vasodilation by NO-dependent and NO-independent actions. AB - Proteinase-activated receptors (PARs) are a novel class of G protein-coupled receptors that respond to signals through endogenous proteinases. PAR activation involves enzymatic cleavage of the extracellular NH(2)-terminal domain and unmasking of a new NH(2) terminus, which serves as an anchored ligand to activate the receptor. At least four PAR subtypes have been identified. In the present study, we used the in vitro perfused hydronephrotic rat kidney to examine the effects of activating PAR-2 on the afferent arteriole. The synthetic peptide SLIGRL-NH(2), which corresponds to the exposed ligand sequence and selectively activates PAR-2, did not alter basal afferent arteriolar diameter but caused a concentration-dependent vasodilation (3-30 microM) of arterioles preconstricted by angiotensin II (0.1 nM). A modified peptide sequence (LSIGRL-NH(2), inactive at PAR-2) had no effect. This vasodilation was characterized by an initial transient component followed by a smaller sustained response. A similar pattern of vasodilation was seen when SLIGRL-NH(2) was administered to isolated perfused normal rat kidney. The sustained component of the PAR-2-induced afferent arteriolar vasodilation was eliminated by nitric oxide (NO) synthase inhibition (100 microM nitro-L-arginine methyl ester). In contrast, the transient vasodilation persisted under these conditions. This transient response was not observed when afferent arterioles were preconstricted with elevated KCl, suggesting involvement of an endothelium-derived hyperpolarizing factor. Finally, RT-PCR revealed the presence of PAR-2 mRNA in isolated afferent arterioles. These findings indicate that PAR-2 is expressed in the afferent arteriole and that its activation elicits afferent arteriolar vasodilation by NO-dependent and NO independent mechanisms. PMID- 11934702 TI - TGF-beta-induced Ca(2+) influx involves the type III IP(3) receptor and regulates actin cytoskeleton. AB - Ca(2+) influx has been postulated to modulate the signaling pathway of transforming growth factor-beta (TGF-beta); however, the underlying mechanism and functional significance of TGF-beta-induced stimulation of Ca(2+) influx are unclear. We show here that TGF-beta stimulates Ca(2+) influx in mesangial cells without Ca(2+) release. The influx of Ca(2+) is prevented by pharmacological inhibitors of inositol 1,4,5-trisphosphate receptors (IP(3)R) as well as specific antibodies to type III IP(3)R (IP(3)RIII) but not to type I IP(3)R (IP(3)RI). TGF beta enhances plasma membrane localization of IP(3)RIII, whereas the sarcoplasmic endoplasmic reticulum Ca(2+)-ATPase (SERCA) preferentially translocates to the nucleus. Untreated mesangial cells exhibit actin filamentous protrusions on the cell surface, and treatment with TGF-beta dramatically reduces this pattern. The alterations in the actin cytoskeleton by TGF-beta are dependent on TGF-beta induced Ca(2+) influx. These studies identify a novel pathway by which TGF-beta regulates Ca(2+) influx and induces cytoskeletal alterations. PMID- 11934703 TI - Ca(2+) influx through the osteoclastic plasma membrane ryanodine receptor. AB - We predict that the type 2 ryanodine receptor isoform (RyR-2) located in the osteoclastic membrane functions as a Ca(2+) influx channel and as a divalent cation (Ca(2+)) sensor. Cytosolic Ca(2+) measurements revealed Ca(2+) influx in osteoclasts at depolarized membrane potentials. The cytosolic Ca(2+) change was, as expected, not seen in Ca(2+)-free medium and was blocked by the RyR modulator ryanodine. In contrast, at basal membrane potentials (approximately 25 mV) ryanodine triggered extracellular Ca(2+) influx that was blocked by Ni(2+). In parallel, single-channel recordings obtained from inside-out excised patches revealed a divalent cation-selective approximately 60-pS conductance in symmetric solutions of Ba-aspartate [Ba-Asp; reversal potential (E(rev)) approximately 0 mV]. In the presence of a Ba(2+) gradient, i.e., with Ba-Asp in the pipette and Na-Asp in the bath, channel conductance increased to approximately 120 pS and E(rev) shifted to 21 mV. The conductance was tentatively classified as a RyR gated Ca(2+) channel as it displayed characteristic metastable states and was sensitive to ruthenium red and a specific anti-RyR antibody, Ab(34). To demonstrate that extracellular Ca(2+) sensing occurred at the osteoclastic surface rather than intracellularly, we performed protease protection assays using pronase. Preincubation with pronase resulted in markedly attenuated cytosolic Ca(2+) signals triggered by either Ni(2+) (5 mM) or Cd(2+) (50 microM). Finally, intracellular application of antiserum Ab(34) potently inhibited divalent cation sensing. Together, these results strongly suggest the existence of 1) a membrane-resident Ca(2+) influx channel sensitive to RyR modulators; 2) an extracellular, as opposed to intracellular, divalent cation activation site; and 3) a cytosolic CaM-binding regulatory site for RyR. It is likely therefore that the surface RyR-2 not only gates Ca(2+) influx but also functions as a sensor for extracellular divalent cations. PMID- 11934704 TI - Role of connective tissue growth factor in fibronectin expression and tubulointerstitial fibrosis. AB - Connective tissue growth factor (CTGF) is one of the candidate factors mediating downstream events of transforming growth factor-beta (TGF-beta), but its role in fibrogenic properties of TGF-beta and in tubulointerstitial fibrosis has not yet been clarified. Using unilateral ureteral obstruction (UUO) in rats, we analyzed gene expression of TGF-beta1, CTGF, and fibronectin. We further investigated the effect of blockade of endogenous CTGF on TGF-beta-induced fibronectin expression in cultured rat renal fibroblasts by antisense oligodeoxynucleotide (ODN) treatment. After UUO, CTGF mRNA expression in the obstructed kidney was significantly upregulated subsequent to TGF-beta1, followed by marked induction of fibronectin mRNA. By in situ hybridization, CTGF mRNA was detected mainly in the interstitial fibrotic areas and tubular epithelial cells as well as in parietal glomerular epithelial cells in the obstructed kidney. The interstitial cells expressing CTGF mRNA were also positive for alpha-smooth muscle actin. CTGF antisense ODN transfected into cultured renal fibroblasts significantly attenuated TGF-beta-stimulated upregulation of fibronectin mRNA and protein compared with control ODN transfection, together with inhibited synthesis of type I collagen. With the use of a reporter assay, rat fibronectin promoter activity was increased by 2.5-fold with stimulation by TGF-beta1, and this increase was abolished with antisense CTGF treatment. Thus CTGF plays a crucial role in fibronectin synthesis induced by TGF-beta, suggesting that CTGF blockade could be a possible therapeutic target against tubulointerstitial fibrosis. PMID- 11934705 TI - alpha(2B)-Adrenergic receptors activate MAPK and modulate proliferation of primary cultured proximal tubule cells. AB - In the rat proximal tubule, the alpha(2B)-adrenergic receptor (alpha(2B)-AR) enhances Na(+) reabsorption by increasing the activity of Na(+)/H(+) exchanger isoform NHE3. The mechanisms involved are unclear, and inhibition of cAMP production remains controversial. In this study, we reinvestigated alpha(2B)-AR signaling pathways using rat proximal tubule cells (PTC) in primary culture and LLC-PK(1) cells permanently transfected with the RNG gene (rat nonglycosylated alpha(2)-AR). Binding experiments indicated that PTC express substantial amounts of alpha(2B)-AR (130 fmol/mg protein), and only RNG transcripts were detected. In both cell types, the alpha(2B)-AR is coupled to G protein, and its stimulation by dexmedetomidine, but not by UK-14304, provoked a significant inhibition of the accumulation of cAMP induced by forskolin or parathyroid hormone. Exposure to alpha(2)-agonists increased arachidonic acid release and caused extracellular signal-regulated kinase (ERK)1/2 phosphorylation, which correlated with enhanced mitogen-activated protein kinse (MAPK) activity and nuclear translocation. MAPK phosphorylation was blunted by pertussis toxin but not by protein kinase C desensitization, and it coincided with transient phosphorylation of Shc. Finally, treatment with UK-14304 accelerated cell growth. Further studies will be necessary to clarify the precise mechanism of MAPK activation, but the present data suggest that alpha(2B)-AR may play a positive role during tubular regeneration. PMID- 11934706 TI - Identification of developmentally regulated mesodermal-specific transcript in mouse embryonic metanephros. AB - Mesodermal-specific cDNA or transcript (MEST) was identified by suppression subtractive hybridization-PCR of cDNA isolated from embryonic day 13 vs. newborn mice kidneys. At day 13 of mouse gestation, a high expression of MEST, with a single approximately 2.7-kb transcript that was exclusively localized to the metanephric mesenchyme was observed. The MEST mRNA expression gradually decreased during the later stages and then abruptly decreased in the newborn kidneys and subsequent postnatal life, after which a very mild expression persisted in the glomerular mesangium. Regression in mRNA expression during embryonic renal development appears to be related to methylation of the MEST gene. Treatment of metanephroi, harvested at day 13 of gestation with MEST-specific antisense oligodeoxynucleotide resulted in a dose-dependent decrease in the size of the explants and the nephron population. This was associated with a selective decrease in MEST mRNA expression and accelerated apoptosis of the mesenchyme. These findings suggest that MEST, a gene with a putative mesenchymal cell-derived protein, conceivably plays a role in mammalian metanephric development. PMID- 11934707 TI - Lung inflammation early in cystic fibrosis: bugs are indicted, but the defense is guilty. PMID- 11934708 TI - On the contagious nature of tuberculosis (continued). PMID- 11934709 TI - Sleep apneas: an oxidative stress? PMID- 11934710 TI - Pharmacogenetics of asthma. PMID- 11934711 TI - Ventilator-associated pneumonia. AB - Ventilator-associated pneumonia (VAP) continues to complicate the course of 8 to 28% of patients receiving mechanical ventilation (MV). In contrast to infections of more frequently involved organs (e.g., urinary tract and skin), for which mortality is low, ranging from 1 to 4%, the mortality rate for VAP ranges from 24 to 50% and can reach 76% in some specific settings or when lung infection is caused by high-risk pathogens. The predominant organisms responsible for infection are Staphylococcus aureus, Pseudomonas aeruginosa, and Enterobacteriaceae, but etiologic agents widely differ according to the population of patients in an intensive care unit, duration of hospital stay, and prior antimicrobial therapy. Because appropriate antimicrobial treatment of patients with VAP significantly improves outcome, more rapid identification of infected patients and accurate selection of antimicrobial agents represent important clinical goals. Our personal bias is that using bronchoscopic techniques to obtain protected brush and bronchoalveolar lavage specimens from the affected area in the lung permits physicians to devise a therapeutic strategy that is superior to one based only on clinical evaluation. When fiberoptic bronchoscopy is not available to physicians treating patients clinically suspected of having VAP, we recommend using either a simplified nonbronchoscopic diagnostic procedure or following a strategy in which decisions regarding antibiotic therapy are based on a clinical score constructed from seven variables. Selection of the initial antimicrobial therapy should be based on predominant flora responsible for VAP at each institution, clinical setting, information provided by direct examination of pulmonary secretions, and intrinsic antibacterial activities of antimicrobial agents and their pharmacokinetic characteristics. Further trials will be needed to clarify the optimal duration of treatment and the circumstances in which monotherapy can be safely used. PMID- 11934712 TI - Inflammation, infection, and pulmonary function in infants and young children with cystic fibrosis. AB - Our aim was to study the effect of lower airway infection on clinical parameters, pulmonary function tests, and inflammation in clinically stable infants and young children with cystic fibrosis (CF). To accomplish this goal, a prospective cohort of screened CF patients under 4 years of age were studied, using elective anesthesia and intubation for: passive respiratory mechanics (single breath occlusion passive deflation) and lung volumes (nitrogen washout), under neuromuscular blockade; and bronchoalveolar lavage (BAL) of 3 main bronchi for cytology, cytokine interleukin (IL)-8, and quantitative microbiology. There were 22 children studied, with a mean age of 23.2 months (6.7-44 months). A greater relative risk of lower airway pathogens was associated with prior respiratory admission (3.60, 95% confidence interval [CI] 2.87-4.51), history of asthma (1.75, 95% CI 1.52-2.03), and chronic symptoms (1.50, 95% CI 1.23-1.83), especially wheeze (1.88, 95% CI 1.61-2.19). Lower respiratory pathogens (> or = 10 cfu/ml BAL) were found in 14 out of 22, and greater than 10(5) cfu/ml in 8 out of 22 subjects. The level of pathogens in BAL (log10 cfu/ml) explained 78% of the variability in percent neutrophils and 34% of the variability in IL-8 levels. Pathogen level also correlated with pulmonary function tests of specific respiratory system compliance (r -0.49, p = 0.02) and functional residual capacity over total lung capacity (r 0.49, p = 0.03). We conclude that the presence of pathogens in the lower airways correlated with levels of inflammation, respiratory system compliance, and degree of air trapping. PMID- 11934713 TI - Endotoxin activity and inflammatory markers in the airways of young patients with cystic fibrosis. AB - Chronic endobronchial infection frequently caused by gram-negative organisms and an increased, neutrophil-dominated inflammation are characteristics of cystic fibrosis (CF). The present study examines endotoxin levels in bronchoalveolar lavage fluids of CF versus non-CF (N) control children, and correlates these with the inflammatory markers interleukin-8 and neutrophils. Fifty-five patients with CF and 56 patients without CF between the ages of 0.04 to 13.25 years were included. Infection, defined as a bacterial count above 50,000 cfu/ml, was present in 27 CF and 25 N patients. Endotoxin levels were not different between patients with and without CF (infected: 74.9 +/- 12.1 EU/ml versus 51.4 +/- 12.5 EU/ml, p = 0.16; noninfected: 5.9 +/- 4.8 EU/ml versus 11.1 +/- 4.3 EU/ml, p = 0.28). Endotoxin activity correlated to the number of gram-negative organisms in CF and N patients, and endotoxin activity per bacterial colony forming unit did not differ with various gram-negative species. Both interleukin-8 and neutrophils were positively correlated with endotoxin, but this slope was shifted toward higher levels of inflammation in CF patients. We conclude that it is unlikely that higher levels of endotoxin in the absence of viable bacteria explain the increased inflammatory response in CF. PMID- 11934714 TI - Intracutaneous tests with recombinant allergens in cystic fibrosis patients with allergic bronchopulmonary aspergillosis and Aspergillus allergy. AB - Allergic bronchopulmonary aspergillosis (ABPA), an intensive inflammatory reaction to Aspergillus fumigatus, can cause irreversible lung damage in patients with cystic fibrosis (CF). The aim of this study was to assess if intracutaneous testing with recombinant A. fumigatus allergens (rAsp f ) allowed a reliable diagnosis of ABPA. Fifty patients with CF were tested, 12 suffering from ABPA, 21 with allergy to A. fumigatus, and 17 CF control patients not sensitized to A. fumigatus. All patients with ABPA reacted to at least one of the two intracellular A. fumigatus allergens rAsp f 4, a 30-kD protein of unknown biologic function, and rAsp f 6, a 23-kD manganese superoxide dismutase, at a concentration of 10(-2) microg/ml. The intracutaneous tests were negative or only marginally positive in the patients with allergy to A. fumigatus and completely negative in the CF control patients. The differential responses to the recombinant A. fumigatus allergens were in perfect agreement with our previous serologic results, so that rAsp f 4 and rAsp f 6 can be considered specific markers for ABPA. Early diagnosis of the disease might help to prevent irreversible lung damage and minimize possible steroid-mediated side effects as a consequence of an optimized control of the disease. PMID- 11934715 TI - Acute effects of aerosolized S-nitrosoglutathione in cystic fibrosis. AB - S-nitrosoglutathione (GSNO), a naturally occurring constituent of airway lining fluid, enhances ciliary motility, relaxes airway smooth muscle, inhibits airway epithelial amiloride-sensitive sodium transport, and prevents pathogen replication. Remarkably, airway levels of GSNO are low in patients with cystic fibrosis (CF). We hypothesized that replacement of airway GSNO would improve gas exchange in CF. In a double-blind, placebo controlled study, we administered 0.05 ml/kg of 10 mM GSNO or phosphate buffered saline by aerosol to patients with CF and followed oxygen saturation, spirometry, respiratory rate, blood pressure, heart rate, and expired nitric oxide (NO). Nine patients received GSNO and 11 placebo. GSNO inhalation was associated with a modest but sustained increase in oxygen saturation at all time points. Expired NO increased in the low ppb range with GSNO treatment, peaking at 5 minutes but remaining above baseline at 30 minutes. There were no adverse effects. We conclude that GSNO is well tolerated in patients with CF and improves oxygenation through a mechanism that may be independent of free NO. Further, GSNO breakdown increases expired NO. We suggest that therapy aimed at restoring endogenous GSNO levels in the CF airway may merit study. PMID- 11934716 TI - Delay in diagnosis among hospitalized patients with active tuberculosis- predictors and outcomes. AB - Delayed diagnosis of active pulmonary tuberculosis (TB) among hospitalized patients is common and believed to contribute significantly to nosocomial transmission. This study was conducted to define the occurrence, associated patient risk factors, and outcomes among patients and exposed workers of delayed diagnosis of active pulmonary TB. Among 429 patients newly diagnosed to have active pulmonary TB between June 1992 and June 1995 in 17 acute-care hospitals in four Canadian cities, initiation of appropriate treatment was delayed 1 week or more in 127 (30%). This was associated with atypical clinical and demographic patient characteristics, and after adjustment for these characteristics, with admission to hospitals with low TB admission rate of 0.2-3.3 per 10,000 admissions (odds ratio [OR]: 7.4; 95% confidence interval [CI]: 3.2,17.5) or intermediate TB admissions of 3.4-9.9/10,000 (OR: 2.3; CI: 1.6,3.2) as well as potentially preventable (late) intensive care unit admission (OR: 16.8; CI: 2.0,144) and death (OR: 3.3; CI: 1.7,6.5]). In hospitals with low TB admission rates, initially missed diagnosis, smear-positive patients undergoing bronchoscopy, late intensive care unit admission (OR: 2.3; CI: 0.1,56), and death (OR: 3.8; CI: 1.2,12.1) were more common than in hospitals with high TB admissions (> 10/ 10,000); a similar trend was seen in hospitals with intermediate TB admissions. Even after adjustment for workers' characteristics and ventilation in patients' rooms tuberculin conversions were disproportionately high in hospitals with low and intermediate TB admission rates and significantly higher in hospitals with overall TB mortality rate above 10% (OR: 2.5; CI: 1.6,3.7). In the hospitals studied, as the rate of TB admissions decreased, the likelihood of poor outcomes and risk of transmission of TB infection per hospitalized patient with TB increased. Institutional risk of TB transmission was poorly correlated with number of patients with TB and better correlated with indicators of patient care such as delayed diagnosis and treatment and overall TB related patient mortality. PMID- 11934717 TI - Increased adhesion molecules expression and production of reactive oxygen species in leukocytes of sleep apnea patients. AB - Obstructive sleep apnea (OSA) is associated with increased cardiovascular morbidity and mortality. Free radicals and adhesion molecules were implicated in the pathogenesis of atherosclerosis leading to cardiovascular disorders. Therefore, we investigated the link between CD15, CD11c, CD11b, and CD64 expression on leukocytes and their ability to generate reactive oxygen species (ROS) in patients with OSA and control volunteers. We also studied the effects of hypoxia in vitro on monocytes from control subjects and the ability of monocytes from both groups to adhere to human endothelial cells in culture. The effect of nasal continuous positive airway pressure (nCPAP) treatment was studied as well. We found that OSA was associated with increased expression of adhesion molecules CD15 and CD11c on monocytes, increased adherence of monocytes in culture to human endothelial cells, increased intracellular ROS production in some monocyte and granulocyte subpopulations, and upregulation of CD15 expression due to hypoxia in vitro in monocytes of control subjects. Furthermore, nCPAP treatment was associated with downregulation of CD15 and CD11c monocyte expression and decreased basal ROS production in CD11c+ monocytes. Monocyte adherence to endothelial cells decreased as well. Our findings provide one of the possible mechanisms for explaining the high rate of cardiovascular morbidity in patients with sleep apnea. PMID- 11934718 TI - Diagnosis of sleep apnea by automatic analysis of nasal pressure and forced oscillation impedance. AB - Detecting and differentiating central and obstructive respiratory events is an important aspect of the diagnosis of sleep-related breathing disorders with respect to the choice of an appropriate treatment. The purpose of this study was to evaluate the performance of a new algorithm for automated detection and classification of apneas and hypopneas, compared with visual analysis of standard polysomnographic signals. The algorithm is based on time series analysis of nasal mask pressure and a forced oscillation signal related to mechanical respiratory input impedance, measured at a frequency of 20 Hz throughout the night. The method was applied to all-night measurements on 19 subjects. Two experts in sleep medicine independently scored the corresponding simultaneously recorded polysomnographic signals. Evaluating the agreement between two scorers by a weighted kappa statistic on a second-by-second basis, we found that inter-expert variability and the discrepancy between automatic analysis and visual analysis performed by an expert were not significantly different. Implementation of this algorithm in a device for home monitoring of breathing during sleep might aid in the differential diagnosis of sleep-related breathing disorders and/or as a means for follow-up and treatment control. PMID- 11934719 TI - Genioglossus muscle responsiveness to chemical and mechanical stimuli during non rapid eye movement sleep. AB - Previous studies have suggested that during non-rapid eye movement (NREM) sleep, neither large short-duration resistive loads nor sustained normoxic hypercapnia alone leads to increased genioglossus muscle activation. However, in normal individuals during stable NREM sleep, genioglossus activity rises above baseline as PCO2 rises and airway resistance increases. We therefore hypothesized that combinations of chemical (PCO2, PO2) and mechanical stimuli during NREM sleep would lead to increased genioglossal activation. We studied 15 normal subjects (9 males, 6 females) during stable NREM sleep, measuring genioglossus electromyogram, epiglottic/choanal pressure, and airflow under six conditions: (1) baseline, (2) inspiratory resistive loading (-5 to -15 cm H2O/ L/second), (3) increased PCO2 (5-10 mm Hg above baseline), (4) combined resistive loading and increased PCO2, (5 ) hypoxia (SaO2 80-85%), and (6 ) combined hypoxia/inspiratory resistive loading. Only the combined condition of hypercapnia and resistive loading led to significantly increased genioglossal activation, 3.91 +/- 0.77% to 9.64 +/- 1.96% of maximum. These data suggest that the genioglossus muscle is less responsive to either chemical stimuli (hypercapnia, hypoxia) or inspiratory resistive loading alone during NREM sleep at the degrees tested. When hypercapnia is combined with resistive loading, the muscle does respond. However, the possibility that higher levels of PCO2 or greater resistive loading alone could activate the muscle cannot be excluded. PMID- 11934720 TI - Impaired vasodilator responses in obstructive sleep apnea are improved with continuous positive airway pressure therapy. AB - Obstructive sleep apnea causes cardiovascular morbidity and premature death. Potential links between sleep apnea and cardiovascular complications are chronically elevated activity of the sympathetic nervous system and abnormal vascular function. To explore vascular function, we determined the reactive hyperemic blood flow (RHBF) responses to 10 minutes of forearm arterial occlusion (plethysmography), blood pressure, and muscle sympathetic nerve activity (MSNA, microneurography) in eight patients with sleep apnea and in nine nonapneic control subjects. Peak RHBF and vascular conductance were markedly attenuated in sleep apnea compared with control subjects (p < 0.05). Seven sleep apnea patients were retested after at least two weeks of continuous positive airway pressure (CPAP) therapy. MSNA decreased after CPAP therapy (p < 0.05, n = 6), whereas blood pressure did not change. After CPAP therapy, peak RHBF and vascular conductance were increased compared with before treatment (p < 0.05; n = 7). Thus, vascular function is abnormal in sleep apnea and is improved by CPAP therapy. Furthermore, effective CPAP therapy decreases sympathetic activity in sleep apnea. Thus, sympathoexcitation and abnormal vascular function in patients with sleep apnea appear to be linked to the repetitive nocturnal apneic events. PMID- 11934721 TI - Occupational asthma in greenhouse flower and ornamental plant growers. AB - The objective of this study was to determine the prevalence of occupational asthma and sensitization to workplace allergens in greenhouse flower and/or ornamental plant growers. A random sample of 40 growers who cultivated such crops and had participated in the European Farmers' Study was selected for a cross sectional study that included (1) greenhouse characteristics and air contaminants (dust, endotoxin, and microorganisms), (2) respiratory symptoms, (3) occupational asthma diagnosed by bronchial provocation challenge, and (4) sensitization to workplace flowers or molds. Associations among respiratory symptoms, sensitization, greenhouse characteristics, and air contaminants were analyzed by logistic regression modeling. Thirty-nine growers agreed to participate (response rate, 97.5%). Bronchial provocation challenge confirmed occupational asthma in three workers (7.7%), all of them sensitized to workplace flowers or molds. No cases of occupational asthma were found among nonsensitized growers. Poor ventilation proved to be a marginal risk factor for wheezing (air velocity: odds ratio, 0.11; 95% confidence interval, 0.01-1.04). Sensitization to flowers or molds was found in 13 of 38 growers (34.2%) but was not related to greenhouse characteristics. We conclude that one-fourth of greenhouse flower and/or ornamental plant growers sensitized to workplace flowers or molds have occupational asthma, a disease that is suffered by 8% of the growers who cultivate these crops. PMID- 11934722 TI - The role of indoor allergen sensitization and exposure in causing morbidity in women with asthma. AB - Longitudinal evidence that indoor allergen exposure causes morbidity in sensitized individuals with asthma is scarce. We evaluated the association of allergen sensitization and home exposure to short- and long-term morbidity in 140 women with asthma and to asthma prevalence in 458 women from metropolitan Boston. Cockroach (Blattella germanica), dust mite (Dermatophagoides farinae), and cat (Felis domesticus) allergens in home dust samples, and specific immunoglobulin E antibodies were measured at outset, and doctor-diagnosed asthma and markers of asthma morbidity were ascertained by questionnaire during a 4-year follow-up. Cat and cockroach-sensitive (immunoglobulin E immunocap [Cap] class > or = 1) women with asthma reported greater morbidity in the past year at the start, and during follow-up, if high levels of the relevant allergen were found. Women with asthma sensitized to cat allergen and with concentrations at 8 microg/g or greater were more likely to have used steroid (adjusted odds ratio [95% confidence interval] 2.7 [1.2-6.2]) and wheezed without a cold (odds ratio 6.8 [3.3- 14.0]) during follow-up. Those sensitized and exposed to cockroach (Bla g 1 or 2 > or = 2 U/g) were at least three times more likely to have used steroid and to have attended a hospital emergency room; the size of the effect upon steroid use was maintained, but the precision was reduced and the 95% confidence interval included one (p = 0.07), with adjustment for race and poverty. We conclude that cockroach and cat allergens may contribute to asthma morbidity in sensitized women. PMID- 11934723 TI - Lymphocyte proliferative response to P6 of Haemophilus influenzae is associated with relative protection from exacerbations of chronic obstructive pulmonary disease. AB - Chronic obstructive pulmonary disease (COPD) is characterized by periodic exacerbations, some of which are caused by nontypeable Haemophilus influenzae (NTHI). P6 is an outer membrane lipoprotein that is highly conserved among strains of NTHI. We hypothesized that lymphocytes from patients with COPD who have exacerbations due to NTHI have a decreased ability to recognize P6. The in vitro lymphocyte proliferative response to P6 in 36 patients with COPD and 12 healthy control subjects was studied. Ten patients who had exacerbations due to NTHI in the previous 12 months showed statistically significant lower proliferation to P6 (stimulation index, log transformed mean +/- standard error 0.82 +/- 0.17) compared with 26 patients who had no exacerbations due to NTHI in the previous 12 months (1.42 +/- 0.13) and to 12 healthy control subjects (1.61 +/- 0.16). These three groups had no significant difference in the lymphocyte proliferative response to tetanus toxoid. There was no difference in serum antibody levels to P6 in the two groups with COPD. These results indicate that decreased proliferation of T cells to P6 is associated with exacerbations of COPD and suggest that the ability of T cells to recognize P6 is associated with relative protection from exacerbations due to NTHI. PMID- 11934724 TI - Expiratory asynchrony in proportional assist ventilation. AB - One of the proposed advantages of proportional assist ventilation (PAV) has been the automatic synchrony between the end of the patient's inspiratory effort and the ventilator cycle (i.e., expiratory synchrony). However, recent clinical studies have shown a prolonged ventilator inspiratory time or even a "runaway" phenomenon with the normal use of PAV. We hypothesize that control-system delay may account for this, because in reality there is always some degree of delays between control-system's input and output in all ventilators. Computer simulation study to date has not taken into account the potential effect of control-system delay on expiratory synchrony. We therefore created a computer model in which the parameter of control-system delay time was introduced. We found that significant expiratory asynchrony may occur with this more realistic model of PAV. The ventilator flow termination may fall behind the completion of the patient inspiration by as long as 0.33 seconds under the selected simulation conditions. The inspiratory termination delay time is in proportion to the control-system delay time, the respiratory time constant, and the assist gain settings. In conclusion, this model indicates that due to the unavoidable control-system delay in the ventilators, expiratory asynchrony may be an inherent shortcoming associated with PAV. PMID- 11934725 TI - Airway pressures and early barotrauma in patients with acute lung injury and acute respiratory distress syndrome. AB - The determinants of barotrauma in mechanically ventilated patients with acute lung injury or acute respiratory distress syndrome (ALI/ARDS) have not been clearly established. Using data from ARDS Network randomized controlled trials, we retrospectively examined the association between airway pressures and the risk of early barotrauma in a cohort of 718 patients with ALI/ARDS and no baseline barotrauma. We studied airway pressures at three exposure intervals: baseline, one day preceding the barotrauma event (one-day lag), and concurrent with the barotrauma event. During the first four study days, the cumulative incidence of barotrauma was 13% (95% confidence interval [CI] 10.6 to 15.6%). In a forward stepwise Cox proportional hazards analysis using time-dependent variables, higher concurrent positive end-expiratory pressure (PEEP) was associated with an increased risk of early barotrauma (relative hazard [RH] 1.67 per 5-cm H2O increment; 95% CI 1.35-2.07). Once concurrent PEEP was selected into the model, no other airway pressure was related to barotrauma, including plateau pressure. In the multivariate analysis, higher concurrent PEEP was also related to a greater risk of barotrauma (RH 1.93; 95% CI 1.44-2.60), controlling for age, ventilator group (6 versus 12 ml/kg), baseline PEEP, baseline plateau pressure, baseline tidal volume, Acute Physiology and Chronic Health Evaluation score, vasopressor use, serum albumin, hepatic failure, and coagulopathy. When one-day lagged values of PEEP were analyzed, higher PEEP was associated with a greater risk of barotrauma (RH 1.38 per 5-cm H2O increment; 95% CI 1.09-1.76). Controlling for the covariates, higher PEEP was related to an increased risk of barotrauma (RH 1.50; 95% CI 0.98- 2.30). In conclusion, higher PEEP may increase the likelihood of early barotrauma in ALI/ARDS. PMID- 11934727 TI - Increased levels of antimicrobial peptides in tracheal aspirates of newborn infants during infection. AB - Pneumonia and systemic infection are common in premature infants. The antimicrobial peptides human beta-defensin 1 and 2 (hBD-1 and hBD-2) and the cathelicidin LL-37/hCAP-18 are effector molecules of the innate respiratory immune system. It is unknown whether these host defense substances are produced in the respiratory tract of newborns. Concentrations of these peptides were determined in tracheal aspirates of mechanically ventilated newborn infants. All three antimicrobial peptides could be detected in airway lining fluid with equivalent levels in term and preterm newborns. Concentrations of antimicrobial peptides correlated with each other and with levels of interleukin-8 and tumor necrosis factor-alpha in the bronchoalveolar lavage fluid. Pulmonary or systemic infections were associated with significantly increased concentrations of LL-37, hBD-1, and hBD-2. Western blotting detected mature peptides in the lavage fluid. In conclusion, mucosal antimicrobial peptides are present in lung secretions of premature and mature newborns. The molecules are upregulated in response to infection and inflammation and probably represent effector molecules of the respiratory defense system. PMID- 11934726 TI - Prolonged methylprednisolone treatment suppresses systemic inflammation in patients with unresolving acute respiratory distress syndrome: evidence for inadequate endogenous glucocorticoid secretion and inflammation-induced immune cell resistance to glucocorticoids. AB - Nuclear factor-kappaB (NF-kappaB) and glucocorticoid receptor-alpha (GR-alpha) have diametrically opposed functions in regulating inflammation. We investigated whether unresolving acute respiratory distress syndrome (ARDS) is associated with systemic inflammation- induced glucocorticoid resistance and whether prolonged methylprednisolone administration accelerates the suppression of systemic inflammatory indices and normalizes the sensitivity of the immune system to glucocorticoids. Patients enrolled into a randomized trial evaluating prolonged methylprednisolone administration in unresolving ARDS had serial plasma samples collected before and after randomization. In the plasma, we measured the concentrations of tumor necrosis factor-alpha (TNF-alpha), interleukins (IL) IL 1beta and IL-6, adrenocorticotropic hormone (ACTH), and cortisol. The ability of patient plasma to influence the NF-kappaB and GR-signal transduction systems of normal peripheral blood leukocytes (PBL) was examined. Patients treated with methylprednisolone had progressive and sustained reductions of TNF-alpha, IL 1beta, IL-6, ACTH, and cortisol concentrations over time. Normal PBL exposed to plasma samples collected during methylprednisolone exhibited significant progressive increases in all aspects of GR-mediated activity and significant reductions in NF-kappaB DNA-binding and transcription of TNF-alpha and IL-1beta. These findings provide support for the presence of endogenous glucocorticoid inadequacy in the control of inflammation and systemic inflammation-induced peripheral glucocorticoid resistance in ARDS. Prolonged methylprednisolone administration accelerated the resolution of both systemic inflammation and peripheral acquired glucocorticoid resistance in ARDS. PMID- 11934728 TI - Nuclear factor-kappaB activation in alveolar macrophages requires IkappaB kinase beta, but not nuclear factor-kappaB inducing kinase. AB - Cytokine mediated activation of alveolar macrophages (AMs) is an important event in the pathogenesis of fibrosing alveolitis (FA). Through membrane-associated antigens, cytokines (e.g., tumor necrosis-factor-alpha and interleukin-1) are believed to activate a common kinase cascade that initiates the cytoplasmic degradation of IkappaB and nuclear translocation of "nuclear factor-kappaB" (NF kappaB). In the nucleus, NF-kappaB promotes the transcription of genes encoding chemokines and cytokines involved in chronic inflammation. Preventing cytokine mediated NF-kappaB activation is a potential strategy for attenuating the lung injury that occurs in FA. Previously, we have demonstrated that, unlike AMs from healthy volunteers, AMs from patients with inflammatory lung diseases express the coxsackie/adenovirus receptor and the alphav integrins required for adenovirus (Adv) infection. This property allows Adv-mediated transgene delivery to diseased, but not normal, AMs and analysis of molecular pathways involved in gene transcription. In this study, AMs were infected with Adv constructs expressing a defective beta subunit of IkappaB kinase (AdvIKKbetakd) and a defective NF-kappaB inducing kinase (AdvNIKkd) to investigate the contribution of these molecules to NF-kappaB activation. We observed that IKKbeta, but not NIK, was required for NF kappaB activation. The results of this study identify IKKbeta, but not NIK, as a potential therapeutic target in diseases that involve NF-kappaB-dependent gene transcription. PMID- 11934729 TI - Expression of ST2 in helper T lymphocytes of malignant pleural effusions. AB - The objective of this study was to test the hypothesis that accumulated helper T lymphocytes in malignant pleural effusions may shift to T-helper type 2 (Th2) and produce soluble ST2 protein. We took samples of serum and pleural effusions (p-) from patients with carcinomatous pleurisy (CA, n = 17), tuberculous pleurisy (TB, n = 8), and congestive heart failure (HF, n = 5) and compared the concentration of cytokines or ST2. Ex vivo production of interleukin (IL)-4 and IL-10, though not that of interferon (IFN)-gamma or IL-12, from CD4+ T cells isolated from pleural effusions was higher in the CA group than in the TB or HF group. The p ST2 concentrations were significantly higher in the CA group than in the TB or HF group, positively correlated with the percentage of pleural effusion CD4+ T cells (r = 0.432, p = 0.016) and inversely correlated with p-IFN-gamma concentrations (r = -0.423, p = 0.019). Furthermore, mRNA expression of ST2 in CD4+ T cells isolated from group CA was upregulated, compared with that in those isolated from the TB group. These results suggest that CD4+ T cells in CA shift to Th2, which can produce soluble ST2 protein, resulting in increased concentrations of p-ST2 in malignant pleural effusion. PMID- 11934730 TI - Clinical relevance of airway 11beta-hydroxysteroid dehydrogenase type II enzyme in asthma. AB - 11beta-hydroxysteroid dehydrogenases (11beta-HSD) are responsible for the conversion of bioactive glucocorticoids to and from inactive metabolites. 11beta HSD2 is generally considered a high-affinity inactivator of natural glucocorticoids, although its activity with synthetic compounds in vivo is unknown. Inhaled corticosteroids (ICS) remain the primary antiinflammatory agents for treating asthma, but little is known about their metabolism in the lung. The aims of this study were to determine whether the 11beta-HSD2 enzyme can be localized to human airway tissue and whether differential expression of this enzyme relates to asthma severity and ICS needs. We studied airway biopsy specimens from 22 asthmatic subjects, in two groups: (1) a group not treated with ICS (n = 7); and (2) a group treated with ICS (range: 200 to 1,500 microg/d; n = 15). A control population consisted of nine nonasthmatic subjects. Immunostaining was done with an immunopurified antibody to human 11beta-HSD2. Immunoreactivity was generally localized to the endothelium of vessels in the lamina propria and to airway epithelium both in asthmatic patients and nonasthmatic controls. There was a statistically significant inverse relationship between the ICS dose required for effective treatment and the extent of epithelial 11beta-HSD2 staining (r = -0.44; p = 0.04). This is consistent with 11beta-HSD2 acting as an oxidoreductase that regenerates rather than inactivates ICS. This study suggests that glucocorticoid sensitivity in the lung is not determined by ICS breakdown, but may be related to 11beta-HSD2 sustaining the activation of synthetic glucocorticoids. PMID- 11934731 TI - Inhibition of antigen-induced eosinophilia and airway hyperresponsiveness by antisense oligonucleotides directed against the common beta chain of IL-3, IL-5, GM-CSF receptors in a rat model of allergic asthma. AB - Airway obstruction, hyperresponsiveness, and the accumulation and persistence within the airways of inflammatory cells characterize asthma. Interleukin (IL)-3, granulocyte macrophage colony- stimulating factor (GM-CSF), and IL-5 are among several cytokines that have been shown to be increased in asthma and to contribute to atopic inflammation. They mediate their effect via receptors that have a common beta subunit (beta(c)). We hypothesized that blocking of this common beta(c) would impair the airway response to antigen. We report that an antisense (AS) phosphorothioate oligodeoxynucleotide (ODN) found to specifically inhibit transcription of the beta(c) in rat bone marrow cells also caused inhibition of beta(c) mRNA expression and of immunoreactive cells within the lungs of Brown Norway (BN) rats when injected intratracheally (p < 0.01). Inhibition of beta(c) significantly reduced (p < 0.01) experimentally induced eosinophilia in vivo in ovalbumin (OVA)-sensitized BN rats after antigen challenge. Furthermore, when compared with mismatch-treated rats, beta(c) AS-ODN caused inhibition of antigen-induced airway hyperresponsiveness to leukotriene D4. Taken together, our findings demonstrate that the common beta(c) of IL-3, IL 5, and GM-CSF receptors is involved in the eosinophil influx and airway hyperresponsiveness that follow OVA challenge and underscore the potential utility of a topical antisense approach targeting beta(c) for the treatment of asthma. PMID- 11934732 TI - Initial therapy in clinical asthma studies often does not equal therapy for mild, persistent asthma. PMID- 11934733 TI - The step 3 dilemma for moderate persistent asthma. PMID- 11934734 TI - Upper airway inflammation in obstructive sleep apnea. PMID- 11934735 TI - Bioinformatics and the theoretical foundations of molecular biology. PMID- 11934736 TI - Separation of nearly identical repeats in shotgun assemblies using defined nucleotide positions, DNPs. AB - An increasingly important problem in genome sequencing is the failure of the commonly used shotgun assembly programs to correctly assemble repetitive sequences. The assembly of non-repetitive regions or regions containing repeats considerably shorter than the average read length is in practice easy to solve, while longer repeats have been a difficult problem. We here present a statistical method to separate arbitrarily long, almost identical repeats, which makes it possible to correctly assemble complex repetitive sequence regions. The differences between repeat units may be as low as 1% and the sequencing error may be up to ten times higher. The method is based on the realization that a comparison of only a part of all overlapping sequences at a time in a data set does not generate enough information for a conclusive analysis. Our method uses optimal multi-alignments consisting of all the overlaps of each read. This makes it possible to determine defined nucleotide positions, DNPs, which constitute the differences between the repeat units. Differences between repeats are distinguished from sequencing errors using statistical methods, where the probabilities of obtaining certain combinations of candidate DNPs are calculated using the information from the multi-alignments. The use of DNPs and combinations of DNPs will allow for optimal and rapid assemblies of repeated regions. This method can solve repeats that differ in only two positions in a read length, which is the theoretical limit for repeat separation. We predict that this method will be highly useful in shotgun sequencing in the future. PMID- 11934737 TI - Assessing the significance of consistently mis-regulated genes in cancer associated gene expression matrices. AB - MOTIVATION: The simplest level of statistical analysis of cancer associated gene expression matrices is aimed at finding consistently up- or down-regulated genes within a given set of tumor samples. Considering the high level of gene expression diversity detected in cancer, one needs to assess the probability that the consistent mis-regulation of a given gene is due to chance. Furthermore, it is important to determine the required sample number that will ensure the meaningful statistical analysis of massively parallel gene expression measurements. RESULTS: The probability of consistent mis-regulation is calculated in this paper for binarized gene expression data, using combinatorial considerations. For practical purposes, we also provide a set of accurate approximate formulas for determining the same probability in a computationally less intensive way. When the pool of mis-regulatable genes is restricted, the probability of consistent mis-regulation can be overestimated. We show, however, that this effect has little practical consequences for cancer associated gene expression measurements published in the literature. Finally, in order to aid experimental design, we have provided estimates on the required sample number that will ensure that the detected consistent mis-regulation is not due to chance. Our results suggest that less than 20 sufficiently diverse tumor samples may be enough to identify consistently mis-regulated genes in a statistically significant manner. AVAILABILITY: An implementation using Mathematica (tm) of the main equation of the paper, (4), is available at www.me.chalmers.se/~mwahde/bioinfo.html. PMID- 11934738 TI - An integrated approach utilizing artificial neural networks and SELDI mass spectrometry for the classification of human tumours and rapid identification of potential biomarkers. AB - MOTIVATION: MALDI mass spectrometry is able to elicit macromolecular expression data from cellular material and when used in conjunction with Ciphergen protein chip technology (also referred to as SELDI-Surface Enhanced Laser Desorption/Ionization), it permits a semi-high throughput approach to be taken with respect to sample processing and data acquisition. Due to the large array of data that is generated from a single analysis (8-10000 variables using a mass range of 2-15 kDa-this paper) it is essential to implement the use of algorithms that can detect expression patterns from such large volumes of data correlating to a given biological/pathological phenotype from multiple samples. If successful, the methodology could be extrapolated to larger data sets to enable the identification of validated biomarkers correlating strongly to disease progression. This would not only serve to enable tumours to be classified according to their molecular expression profile but could also focus attention upon a relatively small number of molecules that might warrant further biochemical/molecular characterization to assess their suitability as potential therapeutic targets. RESULTS: Using a multi-layer perceptron Artificial Neural Network (ANN) (Neuroshell 2) with a back propagation algorithm we have developed a prototype approach that uses a model system (comprising five low and seven high grade human astrocytomas) to identify mass spectral peaks whose relative intensity values correlate strongly to tumour grade. Analyzing data derived from MALDI mass spectrometry in conjunction with Ciphergen protein chip technology we have used relative importance values, determined from the weights of trained ANNs (Balls et al., Water, Air Soil Pollut., 85, 1467-1472, 1996), to identify masses that accurately predict tumour grade. Implementing a three-stage procedure, we have screened a population of approximately 100000-120000 variables and identified two ions (m/z values of 13454 and 13457) whose relative intensity pattern was significantly reduced in high-grade astrocytoma. The data from this initial study suggests that application of ANN-based approaches can identify molecular ion patterns which strongly associate with disease grade and that its application to larger cohorts of patient material could potentially facilitate the rapid identification of validated biomarkers having significant clinical (i.e. diagnostic/prognostic) potential for the field of cancer biology. AVAILIBILITY: Neuroshell 2 is commercially available from ward systems. PMID- 11934739 TI - Analysis of matched mRNA measurements from two different microarray technologies. AB - MOTIVATION: [corrected] The existence of several technologies for measuring gene expression makes the question of cross-technology agreement of measurements an important issue. Cross-platform utilization of data from different technologies has the potential to reduce the need to duplicate experiments but requires corresponding measurements to be comparable. METHODS: A comparison of mRNA measurements of 2895 sequence-matched genes in 56 cell lines from the standard panel of 60 cancer cell lines from the National Cancer Institute (NCI 60) was carried out by calculating correlation between matched measurements and calculating concordance between cluster from two high-throughput DNA microarray technologies, Stanford type cDNA microarrays and Affymetrix oligonucleotide microarrays. RESULTS: In general, corresponding measurements from the two platforms showed poor correlation. Clusters of genes and cell lines were discordant between the two technologies, suggesting that relative intra technology relationships were not preserved. GC-content, sequence length, average signal intensity, and an estimator of cross-hybridization were found to be associated with the degree of correlation. This suggests gene-specific, or more correctly probe-specific, factors influencing measurements differently in the two platforms, implying a poor prognosis for a broad utilization of gene expression measurements across platforms. PMID- 11934740 TI - A mixture model-based approach to the clustering of microarray expression data. AB - MOTIVATION: This paper introduces the software EMMIX-GENE that has been developed for the specific purpose of a model-based approach to the clustering of microarray expression data, in particular, of tissue samples on a very large number of genes. The latter is a nonstandard problem in parametric cluster analysis because the dimension of the feature space (the number of genes) is typically much greater than the number of tissues. A feasible approach is provided by first selecting a subset of the genes relevant for the clustering of the tissue samples by fitting mixtures of t distributions to rank the genes in order of increasing size of the likelihood ratio statistic for the test of one versus two components in the mixture model. The imposition of a threshold on the likelihood ratio statistic used in conjunction with a threshold on the size of a cluster allows the selection of a relevant set of genes. However, even this reduced set of genes will usually be too large for a normal mixture model to be fitted directly to the tissues, and so the use of mixtures of factor analyzers is exploited to reduce effectively the dimension of the feature space of genes. RESULTS: The usefulness of the EMMIX-GENE approach for the clustering of tissue samples is demonstrated on two well-known data sets on colon and leukaemia tissues. For both data sets, relevant subsets of the genes are able to be selected that reveal interesting clusterings of the tissues that are either consistent with the external classification of the tissues or with background and biological knowledge of these sets. AVAILABILITY: EMMIX-GENE is available at http://www.maths.uq.edu.au/~gjm/emmix-gene/ PMID- 11934741 TI - Microarray data warehouse allowing for inclusion of experiment annotations in statistical analysis. AB - MOTIVATION: Microarray technology provides access to expression levels of thousands of genes at once, producing large amounts of data. These datasets are valuable only if they are annotated by sufficiently detailed experiment descriptions. However, in many databases a substantial number of these annotations is in free-text format and not readily accessible to computer-aided analysis. RESULTS: The Multi-Conditional Hybridization Intensity Processing System (M-CHIPS), a data warehousing concept, focuses on providing both structure and algorithms suitable for statistical analysis of a microarray database's entire contents including the experiment annotations. It addresses the rapid growth of the amount of hybridization data, more detailed experimental descriptions, and new kinds of experiments in the future. We have developed a storage concept, a particular instance of which is an organism-specific database. Although these databases may contain different ontologies of experiment annotations, they share the same structure and therefore can be accessed by the very same statistical algorithms. Experiment ontologies have not yet reached their final shape, and standards are reduced to minimal conventions that do not yet warrant extensive description. An ontology-independent structure enables updates of annotation hierarchies during normal database operation without altering the structure. AVAILABILITY AND SUPPLEMENTARY INFORMATION: http://www.dkfz.de/tbi/services/mchips PMID- 11934742 TI - JenPep: a database of quantitative functional peptide data for immunology. AB - MOTIVATION: The compilation of quantitative binding data underlies attempts to derive tools for the accurate prediction of epitopes in cellular immunology and is part of our concerted goal to develop practical computational vaccinology. RESULTS: JenPep is a family of relational databases supporting the growing community of immunoinformaticians. It contains quantitative data on peptide binding to Major Histocompatibility Complexes (MHCs) and to Transmembrane Peptide Transporter (TAP), as well as an annotated list of T-cell epitopes. AVAILABILITY: The database is available via the Internet. An HTML interface allowing searching of the database can be found at the following address: http://www.jenner.ac.uk/JenPep. PMID- 11934743 TI - PatternHunter: faster and more sensitive homology search. AB - MOTIVATION: Genomics and proteomics studies routinely depend on homology searches based on the strategy of finding short seed matches which are then extended. The exploding genomic data growth presents a dilemma for DNA homology search techniques: increasing seed size decreases sensitivity whereas decreasing seed size slows down computation. RESULTS: We present a new homology search algorithm 'PatternHunter' that uses a novel seed model for increased sensitivity and new hit-processing techniques for significantly increased speed. At Blast levels of sensitivity, PatternHunter is able to find homologies between sequences as large as human chromosomes, in mere hours on a desktop. AVAILABILITY: PatternHunter is available at http://www.bioinformaticssolutions.com, as a commercial package. It runs on all platforms that support Java. PatternHunter technology is being patented; commercial use requires a license from BSI, while non-commercial use will be free. PMID- 11934744 TI - D-ASSIRC: distributed program for finding sequence similarities in genomes. AB - MOTIVATION: Locating the regions of similarity in a genome requires the availability of appropriate tools such as 'Accelerated Search for SImilar Regions in Chromosomes' (ASSIRC; Vincens et al., Bioinformatics, 14, 715-725, 1998). The aim of this paper is to present different strategies for improving this program by distributing the operations and data to multiple processing units and to assess the efficiency of the different implementations in terms of running time as a function of the number of processing units. RESULTS: The new version D ASSIRCis based on three alternative strategies of task sharing: (1) a distributed search using the splitting of studied sequences into large overlapping subsequences (strategy ASS); (2) two distributed searches for repeated exact motifs of fixed size either managed by a central processor (strategy AGD) or locally managed by numerous processors (strategy ALD). The result is that the strategy ASSis suitable for a large number of processing units (the time was divided by a factor of 12 when the number of processing units was increased from 1 to 16) wheras the strategy ALDis better for a small set of processors (typically for four or six). The different proposed strategies are efficient for various applications in genomic research, particularly for locating similarities of nucleic sequences in large genomes. AVAILABILITY: D-ASSIRCis freely available by anonymous FTP at ftp://ftp.ens.fr/pub/molbio/dassirc.tar.gz. Sources and binaries for Solaris and Linux are included in the distribution. PMID- 11934746 TI - The complexity of comparing reaction systems. AB - MOTIVATION: As more genomic data becomes available there is increased attention on understanding the mechanisms encoded in the genome. New XML dialects like CellML and Systems Biology Markup Language (SBML) are being developed to describe biological networks of all types. In the absence of detailed kinetic information for these networks, stoichiometric data is an especially valuable source of information. Network databases are the next logical step beyond storing purely genomic information. Just as comparison of entries in genomic databases has been a vital algorithmic problem through the course of the sequencing project, comparison of networks in network databases will be a crucial problem as we seek to integrate higher-order network knowledge. RESULTS: We show that comparing the stoichiometric structure of two reactions systems is equivalent to the graph isomorphism problem. This is encouraging because graph isomorphism is, in practice, a tractable problem using heuristics. The analogous problem of searching for a subsystem of a reaction system is NP-complete. We also discuss heuristic issues in implementations for practical comparison of stoichiometric matrices. PMID- 11934745 TI - Multiple sequence alignment using partial order graphs. AB - MOTIVATION: Progressive Multiple Sequence Alignment (MSA) methods depend on reducing an MSA to a linear profile for each alignment step. However, this leads to loss of information needed for accurate alignment, and gap scoring artifacts. RESULTS: We present a graph representation of an MSA that can itself be aligned directly by pairwise dynamic programming, eliminating the need to reduce the MSA to a profile. This enables our algorithm (Partial Order Alignment (POA)) to guarantee that the optimal alignment of each new sequence versus each sequence in the MSA will be considered. Moreover, this algorithm introduces a new edit operator, homologous recombination, important for multidomain sequences. The algorithm has improved speed (linear time complexity) over existing MSA algorithms, enabling construction of massive and complex alignments (e.g. an alignment of 5000 sequences in 4 h on a Pentium II). We demonstrate the utility of this algorithm on a family of multidomain SH2 proteins, and on EST assemblies containing alternative splicing and polymorphism. AVAILABILITY: The partial order alignment program POA is available at http://www.bioinformatics.ucla.edu/poa. PMID- 11934747 TI - STOCKS: STOChastic Kinetic Simulations of biochemical systems with Gillespie algorithm. AB - MOTIVATION: The availability of a huge amount of molecular data concerning various biochemical reactions provoked numerous attempts to study the dynamics of cellular processes by means of kinetic models and computer simulations. Biochemical processes frequently involve small numbers of molecules (e.g. a few molecules of a transcriptional regulator binding to one 'molecule' of a DNA regulatory region). Such reactions are subject to significant stochastic fluctuations. Monte Carlo methods must be employed to study the functional consequences of the fluctuations and simulate processes that cannot be modelled by continuous fluxes of matter. This provides the motivation to develop software dedicated to Monte Carlo simulations of cellular processes with the rigorously proven Gillespie algorithm. RESULTS: STOCKS, software for the stochastic kinetic simulation of biochemical processes is presented. The program uses a rigorously derived Gillespie algorithm that has been shown to be applicable to the study of prokaryotic gene expression. Features dedicated to the study of cellular processes are implemented, such as the possibility to study a process in the range of several cell generations with the application of a simple cell division model. Taking expression of Escherichia coli beta-galactosidase as an example, it is shown that the program is able to simulate systems composed of reactions varying in several orders of magnitude by means of reaction rates and the numbers of molecules involved. AVAILABILITY: The software is available at ftp://ibbrain.ibb.waw.pl/stocksand http://www.ibb.waw.pl/stocks. SUPPLEMENTARY INFORMATION: Parameters of the model of prokaryotic gene expression are available in example files of software distribution. PMID- 11934748 TI - Long-range control of expression in yeast. PMID- 11934749 TI - Assembly of fingerprint contigs: parallelized FPC. AB - SUMMARY: One of the more common uses of the program FingerPrint Contigs (FPC) is to assemble random restriction digest 'fingerprints' of overlapping genomic clones into contigs. To improve the rate of assembling contigs from large fingerprint databases we have adapted FPC so that it can be run in parallel on multiple processors and servers. The current version of 'parallelized FPC' has been used in our laboratory to assemble mammalian BAC fingerprint databases, each containing more than 300000 BAC fingerprints. AVAILABILITY: This parallelized version of FPC is available under the GNU GPL licence, and can be downloaded from ftp://ftp.bcgsc.bc.ca/pub/fpcd. PMID- 11934750 TI - OligoArray: genome-scale oligonucleotide design for microarrays. AB - SUMMARY: OligoArray is a program that computes gene specific and secondary structure free oligonucleotides for genome-scale oligonucleotide microarray construction or other applications. AVAILABILITY: The program code is distributed under the GNU General Public License and is freely available for non-profit use via request from the authors. PMID- 11934751 TI - ProbeMatchDB--a web database for finding equivalent probes across microarray platforms and species. AB - SUMMARY: ProbeMatchDB is a web-based database designed to facilitate the search of EST/cDNA sequences or STS markers that can be used to represent the same gene across different microarray platforms and species. It can be used for finding equivalent EST clones in the Research Genetics sequence verified clone set based on results from Affymetirx GeneChips. It will also help to identify probes representing orthologous genes across human, mouse and rat on different microarray platforms. AVAILABILITY: The database is accessible at http://brainarray.mhri.med.umich.edu/MARRAY/BC_ASP/brainarray.htm by clicking the 'Query ProbeMatchDB' link. PMID- 11934752 TI - GeneANOVA--gene expression analysis of variance. AB - SUMMARY: GeneANOVA is an ANOVA-based software devoted to the analysis of gene expression data. AVAILABILITY: GeneANOVA is freely available on request for non commercial use. PMID- 11934753 TI - GRIMM: genome rearrangements web server. AB - SUMMARY: Genome Rearrangements In Man and Mouse (GRIMM) is a tool for analyzing rearrangements of gene orders in pairs of unichromosomal and multichromosomal genomes, with either signed or unsigned gene data. Although there are several programs for analyzing rearrangements in unichromosomal genomes, this is the first to analyze rearrangements in multichromosomal genomes. GRIMM also provides a new algorithm for analyzing comparative maps for which gene directions are unknown. AVAILABILITY: A web server, with instructions and sample data, is available at http://www-cse.ucsd.edu/groups/bioinformatics/GRIMM. PMID- 11934754 TI - MaxBench: evaluation of sequence and structure comparison methods. AB - SUMMARY: MaxBench is a web-based system available for evaluating the results of sequence and structure comparison methods, based on the SCOP protein domain classification. The system makes it easy for developers to both compare the overall performance of their methods to standard algorithms and investigate the results of individual comparisons. AVAILABILITY: http://www.sanger.ac.uk/Users/lp1/MaxBench/ PMID- 11934755 TI - CASA: a server for the critical assessment of protein sequence alignment accuracy. AB - SUMMARY: A public server for evaluating the accuracy of protein sequence alignment methods is presented. CASA is an implementation of the alignment accuracy benchmark presented by Sauder et al. (Proteins, 40, 6-22, 2000). The benchmark currently contains 39321 pairwise protein structure alignments produced with the CE program from SCOP domain definitions. The server produces graphical and tabular comparisons of the accuracy of a user's input sequence alignments with other commonly used programs, such as BLAST, PSI-BLAST, Clustal W, and SAM T99. AVAILABILITY: The server is located at http://capb.dbi.udel.edu/casa. PMID- 11934756 TI - BetaTPred: prediction of beta-TURNS in a protein using statistical algorithms. AB - MOTIVATION: beta-turns play an important role from a structural and functional point of view. beta-turns are the most common type of non-repetitive structures in proteins and comprise on average, 25% of the residues. In the past numerous methods have been developed to predict beta-turns in a protein. Most of these prediction methods are based on statistical approaches. In order to utilize the full potential of these methods, there is a need to develop a web server. RESULTS: This paper describes a web server called BetaTPred, developed for predicting beta-TURNS in a protein from its amino acid sequence. BetaTPred allows the user to predict turns in a protein using existing statistical algorithms. It also allows to predict different types of beta-TURNS e.g. type I, I', II, II', VI, VIII and non-specific. This server assists the users in predicting the consensus beta-TURNS in a protein. AVAILABILITY: The server is accessible from http://imtech.res.in/raghava/betatpred/ PMID- 11934757 TI - DIVERGE: phylogeny-based analysis for functional-structural divergence of a protein family. AB - SUMMARY: DetectIng Variability in Evolutionary Rates among GEnes (DIVERGE) is a software system to study functional divergence of a protein family by detecting site-specific change in evolutionary rate using a multiple alignment of amino acid sequences for a given phylogenetic tree. The program first conducts a statistical test for site-specific rate shifts along the tree, and predicting candidate amino acid residues responsible for functional divergence based on posterior analysis. These results can then be mapped on the 3D protein structure if available. AVAILABILITY: DIVERGE is available free of charge from http://xgu1.zool.iastate.edu/. Distribution packages for both Linux and Microsoft Windows operating systems are available, including manual and example files. PMID- 11934758 TI - TREE-PUZZLE: maximum likelihood phylogenetic analysis using quartets and parallel computing. AB - SUMMARY: TREE-PUZZLE is a program package for quartet-based maximum-likelihood phylogenetic analysis (formerly PUZZLE, Strimmer and von Haeseler, Mol. Biol. Evol., 13, 964-969, 1996) that provides methods for reconstruction, comparison, and testing of trees and models on DNA as well as protein sequences. To reduce waiting time for larger datasets the tree reconstruction part of the software has been parallelized using message passing that runs on clusters of workstations as well as parallel computers. AVAILABILITY: http://www.tree-puzzle.de. The program is written in ANSI C. TREE-PUZZLE can be run on UNIX, Windows and Mac systems, including Mac OS X. To run the parallel version of PUZZLE, a Message Passing Interface (MPI) library has to be installed on the system. Free MPI implementations are available on the Web (cf. http://www.lam mpi.org/mpi/implementations/). PMID- 11934759 TI - Areca nut use: an independent risk factor for oral cancer. PMID- 11934760 TI - Murder by fake drugs. PMID- 11934761 TI - Management of co-infection with HIV and TB. PMID- 11934763 TI - Pakistan's doctors protest at killing of 13 colleagues this year. PMID- 11934762 TI - Epidemiology of HIV in China. PMID- 11934764 TI - Staff shortages threaten public health in London. PMID- 11934766 TI - Indian group seeks ban on use of electroconvulsive therapy without anaesthesia. PMID- 11934767 TI - Bush to drop medical records privacy clause. PMID- 11934768 TI - Global fund overwhelmed by requests. PMID- 11934769 TI - HIV drug under review as firm withdraws FDA application. PMID- 11934770 TI - Oregon jury hits Philip Morris with $150m damages for low tar cigarettes. PMID- 11934771 TI - WHO links long term pill use to cervical cancer. PMID- 11934772 TI - Modelling cost effectiveness of meningococcal serogroup C conjugate vaccination campaign in England and Wales. AB - OBJECTIVES: To assess the cost effectiveness of a meningococcal serogroup C conjugate vaccination campaign in 0-17 year olds. DESIGN: Cost effectiveness analysis from the perspective of the healthcare provider. SETTING: England and Wales. MAIN OUTCOME MEASURE: Cost per life year saved. RESULTS: In 1998-9, immediately before the introduction of meningococcal C vaccination, the burden of serogroup C disease was considerable, with an estimated 1137 cases in people aged 0-17 years and at least 72 deaths. The vaccination campaign is estimated to have cost between 126m pound sterling ($180m, 207m) and 241 pound sterling 3m, 395m), depending on the price of the vaccine. Under base case assumptions the cost per life year saved from the vaccination campaign is estimated to be 6259 pound sterling. School based vaccination was more cost effective than general practice based vaccination because of lower delivery costs. Immunisation of infants aged under 1 year was the least cost effective component of the campaign because, although this maximises the life years gained, the three dose schedule required is more expensive than other methods of delivery. Estimates of the cost per life year saved were sensitive to assumptions on the future incidence of disease and the case fatality ratio. CONCLUSIONS: Meningococcal C vaccination is likely to be more cost effective in all age groups when the incidence of disease is high. It is also more cost effective when given to children aged 1-4 (by general practitioners) and to children and young people aged 5-17 years at school than when administered to infants under 12 months of age or young people aged 16-17 years who are not at school. PMID- 11934773 TI - Impact on survival of intensive follow up after curative resection for colorectal cancer: systematic review and meta-analysis of randomised trials. AB - OBJECTIVE: To review the evidence from clinical trials of follow up of patients after curative resection for colorectal cancer. DESIGN: Systematic review and meta-analysis of randomised controlled trials of intensive compared with control follow up. MAIN OUTCOME MEASURES: All cause mortality at five years (primary outcome). Rates of recurrence of intraluminal, local, and metastatic disease and metachronous (second colorectal primary) cancers (secondary outcomes). RESULTS: Five trials, which included 1342 patients, met the inclusion criteria. Intensive follow up was associated with a reduction in all cause mortality (combined risk ratio 0.81, 95% confidence interval 0.70 to 0.94, P=0.007). The effect was most pronounced in the four extramural detection trials that used computed tomography and frequent measurements of serum carcinoembryonic antigen (risk ratio 0.73, 0.60 to 0.89, P=0.002). Intensive follow up was associated with significantly earlier detection of all recurrences (difference in means 8.5 months, 7.6 to 9.4 months, P<0.001) and an increased detection rate for isolated local recurrences (risk ratio 1.61, 1.12 to 2.32, P=0.011). CONCLUSIONS: Intensive follow up after curative resection for colorectal cancer improves survival. Large trials are required to identify which components of intensive follow up are most beneficial. PMID- 11934774 TI - Cross sectional survey of parents' experience and views of the postmortem examination. AB - OBJECTIVE: To describe parents' experience and views of the postmortem examination after the loss of a baby. DESIGN: Cross sectional survey. SETTING: Hospital with a dedicated bereavement counselling service, Newcastle upon Tyne. PARTICIPANTS: 258 women who had attended a bereavement counselling service at the Royal Victoria Infirmary, Newcastle upon Tyne, on at least one occasion after losing a baby during pregnancy or infancy, between October 1996 and October 2000. METHOD: Self completion postal questionnaire incorporating fixed choice and open ended questions. MAIN OUTCOME MEASURES: Number of respondents who were asked if they would agree to a postmortem examination of their baby, and number who agreed to a postmortem examination; reasons for agreeing and not agreeing to a postmortem examination; quality of explanation received; number who regretted their decision to give or withhold consent for a postmortem examination. RESULTS: 166 (64%) respondents completed the questionnaire. Of these, 148 (89%) had been asked to agree to a postmortem examination on their baby and 120/148 of these respondents (81%) agreed, most of whom recognised benefits resulting from the examination. 101/117 (86%) respondents believed the findings had been explained appropriately. Nine (7%) of the 120 respondents who had agreed to a postmortem examination regretted their decision. Of the respondents who refused an examination, four (14%) had regrets about their decision. DISCUSSION: Parents viewed the postmortem examination as a useful and necessary tool in helping to discover the reasons why their baby had died. Simplifying the language used to explain findings may further raise parents' understanding of the value of the postmortem examination and ensure that they are satisfied with it. Medical staff involved in consent for postmortem examinations should be fully trained in how to ask for parental consent, the postmortem examination procedure, and how to explain the findings. PMID- 11934775 TI - Systematic review of whether nurse practitioners working in primary care can provide equivalent care to doctors. AB - OBJECTIVE: To determine whether nurse practitioners can provide care at first point of contact equivalent to doctors in a primary care setting. DESIGN: Systematic review of randomised controlled trials and prospective observational studies. DATA SOURCES: Cochrane controlled trials register, specialist register of trials maintained by Cochrane Effective Practice and Organisation of Care Group, Medline, Embase, CINAHL, science citation index, database of abstracts of reviews of effectiveness, national research register, hand searches, and published bibliographies. INCLUDED STUDIES: Randomised controlled trials and prospective observational studies comparing nurse practitioners and doctors providing care at first point of contact for patients with undifferentiated health problems in a primary care setting and providing data on one or more of the following outcomes: patient satisfaction, health status, costs, and process of care. RESULTS: 11 trials and 23 observational studies met all the inclusion criteria. Patients were more satisfied with care by a nurse practitioner (standardised mean difference 0.27, 95% confidence interval 0.07 to 0.47). No differences in health status were found. Nurse practitioners had longer consultations (weighted mean difference 3.67 minutes, 2.05 to 5.29) and made more investigations (odds ratio 1.22, 1.02 to 1.46) than did doctors. No differences were found in prescriptions, return consultations, or referrals. Quality of care was in some ways better for nurse practitioner consultations. CONCLUSION: Increasing availability of nurse practitioners in primary care is likely to lead to high levels of patient satisfaction and high quality care. PMID- 11934776 TI - Communicating accuracy of tests to general practitioners: a controlled study. AB - OBJECTIVE: To assess the extent to which different forms of summarising diagnostic test information influence general practitioners' ability to estimate disease probabilities. DESIGN: Controlled questionnaire study. SETTING: Three Swiss conferences in continuous medical education. PARTICIPANTS: 263 general practitioners. INTERVENTION: Questionnaire with multiple choice questions about terms of test accuracy and a clinical vignette with the results of a diagnostic test described in three different ways (test result only, test result plus test sensitivity and specificity, test result plus the positive likelihood ratio presented in plain language). MAIN OUTCOME MEASURES: Doctors' knowledge and application of terms of test accuracy and estimation of disease probability in the clinical vignette. RESULTS: The correct definitions for sensitivity and predictive value were chosen by 76% and 61% of the doctors respectively, but only 22% chose the correct answer for the post-test probability of a positive screening test. In the clinical vignette doctors given the test result only overestimated its diagnostic value (median attributed likelihood ratio (aLR)=9.0, against 2.54 reported in the literature). Providing the scan's sensitivity and specificity reduced the overestimation (median aLR=6.0) but to a lesser extent than simple wording of the likelihood ratio (median aLR=3.0). CONCLUSION: Most general practitioners recognised the correct definitions for sensitivity and positive predictive value but did not apply them correctly. Conveying test accuracy information in simple, non-technical language improved their ability to estimate disease probabilities accurately. PMID- 11934777 TI - Explaining risks: turning numerical data into meaningful pictures. PMID- 11934778 TI - ABC of clinical electrocardiography: Acute myocardial infarction-Part I. PMID- 11934780 TI - The medical profession, the public, and the government. PMID- 11934779 TI - Unhappy doctors: what are the causes and what can be done? PMID- 11934781 TI - From guidance to practice: Why NICE is not enough. PMID- 11934782 TI - Legalising active euthanasia and physician assisted suicide. Assisted suicide is not always as easy as suggested. PMID- 11934783 TI - Doctors should not decide on medical and "existential" suffering. PMID- 11934784 TI - Medical education must be rehabilitated in Afghanistan. PMID- 11934785 TI - National screening programme for diabetic retinopathy. Digital image may be better for screening. PMID- 11934786 TI - Infections and risk factors in entrants to Irish prisons. High prevalence of viral and other sexually transmitted diseases was found in Indian prisons. PMID- 11934787 TI - Complaints about advertising of medicines are encouraged. PMID- 11934788 TI - Declarations for new doctors are unnecessary. PMID- 11934789 TI - Continuity would be achieved with patient held records. PMID- 11934790 TI - Hospital responsibilities and communication must be clear. PMID- 11934791 TI - Pattern of some parents' complaints against doctors must be recognised. PMID- 11934792 TI - Investigating doctors' performance can cause problems of consent and confidentiality. PMID- 11934797 TI - Commentary on neostigmine interactions with non steroidal anti-inflammatory drugs by Miranda et al. AB - The article by Miranda et al. presents data showing significant synergism between NEO and certain NSAIDs administered i.p. in the mouse. Their conclusions follow from isobolar analysis accompanied by statistical confidence limits that show significant differences between the additive (expected) and experimental potencies. This commentary discusses the features of the graphical method of analysis and points out other experimental designs and the methods used to analyse them. PMID- 11934798 TI - Neostigmine interactions with non steroidal anti-inflammatory drugs. AB - 1. The common mechanism of action of non-steroidal anti-inflammatory drugs (NSAIDs) is the inhibition of the enzyme cyclo-oxygenase (COX), however, this inhibition is not enough to completely account for the efficacy of these agents in several models of acute pain. 2. It has been demonstrated that cholinergic agents can induce antinociception, but the nature of the interaction between these agents and NSAIDs drugs has not been studied. The present work evaluates, by isobolographic analysis, the interactions between the cholinergic indirect agonist neostigmine (NEO) and NSAIDs drugs, using a chemical algesiometric test. 3. Intraperitoneal (i.p.) or intrathecal (i.t.) administration of NEO and of the different NSAIDs produced dose-dependent antinociception in the acetic acid writhing test of the mouse. 4. The i.p. or i.t. co-administration of fixed ratios of ED(50) fractions of NSAIDs and NEO, resulted to be synergistic or supra additive for the combinations ketoprofen (KETO) and NEO, paracetamol (PARA) and NEO) and diclofenac (DICLO) and NEO administered i.p. However, the same combinations administered i.t. were only additive. In addition, the combinations meloxicam (MELO) and NEO and piroxicam (PIRO) and NEO, administered either i.p. or i.t., were additive. 5. The results suggest that the co-administration of NEO with some NSAIDs (e.g. KETO, PARA or DICLO) resulted in a synergistic interaction, which may provide evidence of supraspinal antinociception modulation by the increased acetylcholine concentration in the synaptic cleft of cholinergic interneurons. The interaction obtained between neostigmine and the NSAIDs could carry important clinical implications. PMID- 11934799 TI - Gastric antisecretory role and immunohistochemical localization of cannabinoid receptors in the rat stomach. AB - 1. The role of cannabinoid (CB) receptors in the regulation of gastric acid secretion was investigated in the rat by means of functional experiments and by immunohistochemistry. 2. In anaesthetized rats with lumen-perfused stomach, the non selective CB-receptor agonist WIN 55,212-2 (0.30 - 4.00 micromol kg(-1), i.v.) and the selective CB(1)-receptor agonist HU-210 (0.03 - 1.50 micromol kg( 1), i.v.), dose-dependently decreased the acid secretion induced by both pentagastrin (30 nmol kg(-1) h(-1)) and 2-deoxy-D-glucose (1.25 mmol kg(-1), i.v.). By contrast, neither WIN 55,212-2 (1 - 4 micromol kg(-1), i.v.) nor HU-210 (0.03 - 1.50 micromol kg(-1), i.v.) did modify histamine-induced acid secretion (20 micromol kg(-1) h(-1)). The selective CB(2)-receptor agonist JWH-015 (3 - 10 micromol kg(-1), i.v.) was ineffective. 3. The gastric antisecretory effects of WIN 55,212-2 and HU-210 on pentagastrin-induced acid secretion were prevented by the selective CB(1)-receptor antagonist SR141716A (0.65 micromol kg(-1), i.v.) and unaffected by the selective CB(2)-receptor antagonist SR144528 (0.65 - 2 micromol kg(-1), i.v.). 4. Bilateral cervical vagotomy and ganglionic blockade with hexamethonium (10 mg kg(-1), i.v., followed by continuous infusion of 10 mg kg(-1) h(-1)) significantly reduced, but not abolished, the maximal inhibitory effect of HU-210 (0.3 micromol kg(-1), i.v.) on pentagastrin-induced acid secretion; by contrast, pretreatment with atropine (1 mg kg(-1), i.v.) did not modify the antisecretory effect of HU-210. 5. Immunoreactivity to the CB(1) receptor was co-localized with that of the cholinergic marker choline acetyltransferase in neural elements innervating smooth muscle, mucosa and submucosal blood vessels of rat stomach fundus, corpus and antrum. In contrast, CB(2) receptor-like immunoreactivity was not observed. 6. These results indicate that gastric antisecretory effects of cannabinoids in the rat are mediated by suppression of vagal drive to the stomach through activation of CB(1) receptors, located on pre- and postganglionic cholinergic pathways. However, the ineffectiveness of atropine in reducing the effect of HU-210 suggests that the release of non cholinergic excitatory neurotransmitters may be regulated by CB(1) receptors. PMID- 11934800 TI - Evidence for sigma-1-like receptors in isolated rat liver mitochondrial membranes. AB - 1. Sigma (sigma) receptors have generated a great deal of interest on the basis of their possible roles in various pathologies, including cytoprotection. Although the exact function of sigma-1 (sigma(1)) receptors is not yet known, their role in the regulation of intracellular Ca(2+) levels and sterol biosynthesis, functions that could be assigned to mitochondria, are the only mechanisms described. 2. Using preparations of purified rat liver and brain mitochondria we demonstrate herein the presence of sigma-like binding sites. [(3)H](+)-pentazocine, a sigma(1) radioligand was used to label these sites. 3. In the liver, [(3)H](+)-pentazocine labelled one class of binding sites with high affinity (K(d)=3 nM), similar to that observed in liver microsomes and synaptic membranes. These sites were located on the outer mitochondrial membranes and displayed high affinity for other sigma(1) ligands namely, haloperidol, ifenprodil, carbetapentane or 1,3-di(2-tolyl)guanidine (DTG). 4. The presence of sigma(1) receptors on liver mitochondria was confirmed using double fluorescence immunostaining. 5. [(3)H](+)-pentazocine binding sites were also found on brain mitochondria but they appeared pharmacologically distinct to the liver ones as [(3)H](+)-pentazocine and typical sigma(1) ligands displayed lower affinities for these sites. Nevertheless, [(3)H](+)-pentazocine binding on both liver and brain mitochondria was modulated by progesterone, a putative endogenous ligand for sigma receptors. 6. Our data demonstrates the presence of [(3)H](+)-pentazocine binding sites with pharmacological characteristics identical to sigma(1) receptors on rat liver mitochondrial membranes. The pharmacological significance of these sites and their role on mitochondrial function remain unknown. PMID- 11934801 TI - Stimulation of Ca(2+) influx through ATP receptors on rat brain synaptosomes: identification of functional P2X(7) receptor subtypes. AB - 1. ATP receptors of the P2X class have previously been identified on autonomic nerve endings and on a limited population of CNS neurons. 2. In the present study P2X receptors on mammalian cortical synaptosomes have been identified by a variety of functional and biochemical studies. In choline buffer ATP analogues caused concentration/time dependent Ca(2+) influx. Relative to the effects caused by ATP, benzoylbenzoyl ATP (BzATP) was about seven times more active than ATP while 2-me-S-ATP and ATPgammaS were much less active. alpha,beta-me- ATP and beta,gamma-me-ATP were virtually inactive. In sucrose buffer, relative to choline buffer, the activity of BzATP was more than doubled while activity in sodium buffer was reduced. Moreover, the P2X antagonists PPADS or Brilliant Blue G both significantly attenuated influx. These observations suggest the presence of P2X receptors on synaptosomes which subserve Ca(2+) influx. This activity profile of the ATP analogues and the response to blocking agents are characteristic of responses of P2X(7) receptors. 3. Influx was unaffected by the VSCC inhibitors omega-CTx-MVIIC and (-) 202 - 791, indicating that ATP induced Ca(2+) influx occurred primarily through P2X receptors. 4. P2X(7) receptor protein was identified by Western blotting and immunohistochemical staining. Purified preparations were devoid of significant concentrations of GFAP or the microglial marker OX-42 but contained greatly enriched amounts of syntaxin and SNAP 25. 5. The various pharmacological and biochemical studies were all consistent with the presence of functional P2X(7) receptors. PMID- 11934802 TI - The protective effect of resveratrols on ischaemia-reperfusion injuries of rat hearts is correlated with antioxidant efficacy. AB - 1. Dietary antioxidants are thought to be beneficial in reducing the incidence of coronary heart disease. In this study, we compared resveratrol and analogues on their antioxidation and free radical scavenging activities to their protective effects on ischaemia-reperfusion induced injuries of rat hearts. 2. Astringinin (3,3',4',5-tetrahydroxystilbene) was shown to be a more potent inhibitor than other analogues against Cu(2+)-induced LDL (low-density lipoprotein) oxidation, as measured by the formation of conjugated diene and TBARS (thiobarbituric acid reactive substance) and by the electrophoretic mobility of the oxidized LDL. 3. Resveratrol (trans-3,4',5-trihydroxystilbene) and astringinin scavenged the stable free radical DPPH (1,1-diphenyl-2-picryl-hydrazyl) with an IC(0.200) of 7.1 and 4.3 microM, respectively. 4. Astringinin has a superoxide anion scavenging activity about 160 fold more potent than resveratrol. 5. After a 30 min global ischemia followed by 2 h reperfusion, astringinin (10 microM) significantly reduced infarct size, superoxide anion production and increased functional recovery of the coronary flow in Langendorff-perfused rat hearts. 6. The result showed there is a positive correlation between the anti-oxidation and cardioprotective activities among these phenolic compounds. Our finding together with the fact that astringinin is more water-soluble than resveratrol suggest that astringinin could potentially be used as an anti-oxidant and cardioprotective agent in biological systems. PMID- 11934803 TI - Inhibition by glucocorticoids of the interleukin-1beta-enhanced expression of the mast cell growth factor SCF. AB - 1. Stem cell factor (SCF) is a major mast cell growth factor that promotes differentiation and chemotaxis of mast cells and inhibits their apoptosis. 2. We evaluated the effect of interleukin (IL)-1beta, a major pro-inflammatory cytokine, on the constitutive expression of SCF and studied the effects of two glucocorticoids, budesonide and dexamethasone, on the IL-1beta-enhanced SCF expression. 3. Human lung fibroblasts in culture were serum-starved for 48 h and treated with IL-1beta, budesonide and/or RU486. SCF cDNA was quantified after total RNA reverse transcription by on-line fluorescent polymerase chain reaction. SCF protein was quantified by ELISA. 4. IL-1beta induced an increase in SCF mRNA (+91% at 2.5 h) and protein production (+32%) by human lung fibroblasts in culture (P<0.001). 5. Budesonide inhibited IL-1beta-induced SCF mRNA expression ( 68%) at 2.5 h and even more so at 10 h (-192%) (P<0.001). The expression of SCF protein also decreased by 3.5-fold at 10 h. Results were similar with dexamethasone. The glucocorticoid antagonist RU486 cancelled the effects induced by the glucocorticoids. 6. Increased SCF mRNA levels were associated with increased stability of this mRNA as measured after treatment with actinomycin D (1.9-fold at 2.5 h). Budesonide decreased this IL-1beta-enhanced stability by about 1.5-fold (P<0.001). 7. We conclude that in 'inflammatory' conditions, mimicked in vitro by IL-1beta, glucocorticoid treatment inhibits expression of the mast cell growth factor SCF. The reduced number and activation of mast cells observed in the bronchi of asthmatic patients treated by glucocorticoids may be due in part to this effect. PMID- 11934804 TI - Pharmacologic and autoradiographic evidence for an up-regulation of kinin B(2) receptors in the spinal cord of spontaneously hypertensive rats. AB - 1. The effects of intrathecally (i.t.) injected kinin B(1) and B(2) receptor agonists and antagonists were measured on mean arterial pressure (MAP) and heart rate (HR) of conscious unrestrained spontaneously hypertensive rats (SHR of 16 weeks old) and age-matched normotensive Wistar Kyoto (WKY). Quantitative in vitro autoradiographic studies were also performed on the thoracic spinal cord of both strains with specific radioligands for B(2) receptors, [(125)I]-HPP-Hoe 140, and B(1) receptors, [(125)I]-HPP-[des-Arg(10)]-Hoe140. 2. Bradykinin (BK) (0.81 - 810 pmol) increased MAP dose-dependently with increases or decreases of HR. The pressor response to BK was significantly greater in SHR. The cardiovascular response to 8.1 pmol BK was reversibly blocked by 81 pmol Hoe 140 (B(2) antagonist) but not by 81 - 810 pmol [des-Arg(10)]-Hoe 140 (B(1) antagonist) in both strains. 3. The B(1) receptor agonist, des-Arg(9)-BK (8100 pmol) produced either no effects or increased MAP with variable effects on HR. These responses were similar in both strains and were reversibly blocked by 81 pmol Hoe 140. Inhibition with 8100 pmol [des-Arg(10)]-Hoe 140 was not specific to B(1) agonist mediated responses. 4. [(125)I]-HPP-Hoe 140 specific binding sites were predominantly located to superficial laminae of the dorsal horn and were significantly higher in SHR. Low levels of [(125)I]-HPP-[des-Arg(10)]-HOE 140 specific binding sites were found in all laminae of both strains. 5. It is concluded that the hypersensitivity of the cardiovascular response to BK is due to an increased number of B(2) receptors in the spinal cord of SHR and that B(1) receptors are unlikely involved in spinal cardiovascular regulation in SHR. PMID- 11934805 TI - Pharmacological profile of PKF242-484 and PKF241-466, novel dual inhibitors of TNF-alpha converting enzyme and matrix metalloproteinases, in models of airway inflammation. AB - 1. TNF-alpha converting enzyme (TACE) and matrix metalloproteinases (MMPs) are believed to play a role in various airway inflammatory disorders. Therefore we have tested the effect of two new inhibitors of TACE/MMPs (PKF242-484, PKF241 466) in models of airway inflammation. 2. PKF242-484 and PKF241-466 inhibited purified MMP-1, -2, -3, -9, -13 and rat collagenase at low nanomolar range. Both compounds inhibited the TNF-alpha release from activated human peripheral blood mononuclear cells with IC(50) values of 56+/-28 and 141+/-100 nM, respectively and had no significant effect on the activation of other human leukocytes, as neither neutrophils and eosinophils oxidative burst nor proliferation or cytokines production by T cells were inhibited in vitro. 3. PKF242-484 and PKF241 466 had beneficial effects in two different murine models of acute lung inflammation in vivo. The influx of neutrophils and lymphocytes into the airways was reduced 3 and 24 h after intranasal LPS challenge. This was accompanied by reduced levels of myeloperoxidase and elastase activities in the bronchoalveolar lavage. Furthermore, a complete inhibition of TNF-alpha release into the airways was observed. In addition, PKF242-484 effectively reduced the influx of neutrophils, eosinophils and lymphocytes in a model of acute allergic lung inflammation. 4. PKF242-484 and PKF241-466 are two novel and potent dual inhibitors of TACE and MMPs, which show activity in in vivo models of lung inflammation. Such compounds could have beneficial effects in airway inflammatory conditions such as asthma and chronic obstructive pulmonary disease. PMID- 11934806 TI - Inhibition of LPS-induced chemokine production in human lung endothelial cells by lipid conjugates anchored to the membrane. AB - 1. In acute respiratory distress syndrome (ARDS) induced by endotoxins, a high production of inflammatory mediators by microvascular lung endothelial cells (LMVEC) can be observed. Activation of cells by endotoxins may result in elevated secretion of phospholipase A(2) (sPLA(2)) which is thought to contribute to tissue damage. The present study was undertaken to investigate the role of sPLA(2) in chemokine production in human lung microvascular endothelial cells (LMVEC) stimulated with the endotoxins lipopolysaccharide (LPS) and lipoteichoic acid (LTA). In particular, we investigated the effects of sPLA(2) inhibitors, specifically, the extracellular PLA(2) inhibitors (ExPLIs), composed of N derivatized phosphatidyl-ethanolamine linked to polymeric carriers, and LY311727, a specific inhibitor of non-pancreatic sPLA(2). 2. ExPLIs markedly inhibited LPS and LTA induced production and mRNA expression of the neutrophile attracting chemokines IL-8, Gro-alpha and ENA-78, as well as of the adhesion molecules ICAM 1 and E-selectin. Concomitantly, ExPLIs inhibited the LPS-induced activation of NF-kappaB by LPS but not its activation by TNF-alpha or IL-1. 3. Endotoxin mediated chemokine production in LMVEC seems not to involve PLA(2) activity, since LPS stimulation was not associated with activation of intracellular or secreted PLA(2). It therefore seems that the inhibitory effect of the ExPLIs was not due to their PLA(2) inhibiting capacity. This was supported by the finding that the LPS-induced chemokine production was not affected by the selective sPLA(2) inhibitor LY311727. 4. It is proposed that the ExPLIs may be considered a prototype of potent suppressors of specific endotoxin-induced inflammatory responses, with potential implications for the therapy of subsequent severe inflammation. PMID- 11934807 TI - Role of protein kinase C-epsilon in the development of kappa-opioid receptor tolerance to U50,488H in rat ventricular myocytes. AB - The role of protein kinase C-epsilon (PKC-epsilon) in the development of kappa opioid receptor (kappa-OR) tolerance to the effects of trans-(+/-)-3,4-dichloro-N methyl-N-(2-[1-pyrrolidinyl]cyclohexyl) (U50,488H), the selective agonist of kappa-OR, was determined in rat ventricular myocytes. Incubation of ventricular myocytes with 1 microM U50,488H for 24 h significantly attenuated the inhibitory effects of 30 microM U50,488H on the electrically-induced [Ca(2+)](i) transient and forskolin-stimulated cyclic AMP accumulation, indicating the development of tolerance to the kappa-OR agonist. Chronic treatment of ventricular myocytes with U50,488H also induced translocation of PKC-epsilon to the particulate fraction. On the other hand, administration of 30 microM U50,488H for 10 min induced translocation of PKC-alpha to the particulate fraction in naive ventricular myocytes, but not in cells pretreated with 1 microM U50,488H for 24 h. In ventricular myocytes incubated for 24 h with 1 microM U50,488H together with 1 microM chelerythrine or 1 microM GF109203X, PKC inhibitors, or 0.1 microM epsilonV1-2 peptide, a selective inhibitor of PKC-epsilon, 30 microM U50,488H still produced the inhibitory effect on the electrically-induced [Ca(2+)](i) transient as it did in naive ventricular myocytes. Chronic treatment of ventricular myocytes with U50,488H and chelerythrine also attenuated the development of tolerance to acute U50,488H on cyclic AMP accumulation. Cells exposed to chelerythrine, GF109203X, or epsilonV1-2 peptide alone did not show an altered [Ca(2+)](i) response to U50,488H. These results indicate that activation of PKC-epsilon is a critical step in the development of tolerance in the kappa OR. PMID- 11934808 TI - Stereoselective transport of hydrophilic quaternary drugs by human MDR1 and rat Mdr1b P-glycoproteins. AB - 1. The present study was performed to evaluate and compare the ability of human MDR1-, and rat Mdr1b- and Mdr2-P-glycoproteins to transport hydrophilic monoquaternary drugs. Transport studies were performed with plasma membrane vesicles isolated from MDR1-, Mdr1b-, or Mdr2-overexpressing insect cells. 2. As model substrates we used the N-methylated derivatives of the diastereomers quinidine and quinine, the monoquaternary compounds N-methylquinidine and N methylquinine. Vincristine, an established MDR1 substrate, was used as a reference. 3. We observed ATP-dependent uptake of all drugs studied into MDR1- and Mdr1b-expressing vesicles. Mdr2 was not able to transport these compounds. MDR1- and Mdr1b-mediated transport was saturable, and could be inhibited by various drugs, including PSC-833. 4. For both MDR1 and Mdr1b the V(max)/K(m) ratios (or clearance) of N-methylquinidine were greater than those determined for N-methylquinine. This stereoselective difference was also evident from differential inhibitory studies with the two isomers. 5. Comparison of normalized clearance indicated that human MDR1 was more effective in transporting the tested substrates than rat Mdr1b. 6. In conclusion, our results demonstrate that MDR1 and Mdr1b, but not Mdr2, are able to transport the monoquaternary model drugs; both MDR1 and Mdr1b display stereospecificity for these cations; and indicate human MDR1 is more efficient in transporting these cations than its rat orthologue Mdr1b. PMID- 11934809 TI - 17-beta-oestradiol-induced vasorelaxation in vitro is mediated by eNOS through hsp90 and akt/pkb dependent mechanism. AB - 1. The L-arginine-NO pathway has been implicated in the vasorelaxant effect of 17 beta-oestradiol. Here we have addressed the involvement of two distinct activation steps of endothelial nitric oxide synthase (eNOS) in the 17-beta oestradiol-induced vasorelaxant effect on rat aortic rings. 2. Rat aortic rings contracted with phenylephrine (PE) 1 microM relaxed in a concentration related fashion to 17-beta-oestradiol water soluble cyclodextrin-encapsulated (E2) only when endothelium was present. The pure anti-oestrogen of E2 receptor ICI 182,780 (20 microM) significantly inhibited E2-induced vasorelaxation. 3. Geldanamycin (10 microM), a specific inhibitor of heat shock protein 90 (hsp90) and N(omega) nitro-L-arginine-methyl ester (L-NAME, 100 microM), a nitric oxide synthase inhibitor, significantly inhibited E2-induced vasorelaxation. 4. Incubation of rat aortic rings up to 6 h with LY 294002 (25 microM), a specific inhibitor of PI(3)K akt/pkb pathway reduced E2-induced vasorelaxation. 5. Incubation of rat isolated aorta with E2, induced prostacyclin (PGI(2)) release. PGI(2) levels, measured as 6-keto PGF(1alpha), were abolished by ibuprofen (10 microM), both L NAME and GA did not influence basal or E2-stimulated PGI(2) confirming the specificity of these two compounds on eNOS pathway. 6. In conclusion, we demonstrate that E2 interaction with its receptor is followed by a vasorelaxant effect in rat aortic rings mediated by eNOS activation through both hsp90 and akt/pkb dependent mechanisms. PMID- 11934810 TI - Neuropeptide Y increases 4-aminopyridine-sensitive transient outward potassium current in rat ventricular myocytes. AB - 1. The modulation of 4-aminopyridine sensitive transient outward potassium current (4-AP I(to)) by neuropeptide Y (NPY) (100 nM) in rat ventricular myocytes was examined using the whole cell voltage-clamp technique. 2. Continuous exposure to NPY (100 nM) for 3 - 6 h significantly increased 4-AP I(to) density. The stimulation of 4-AP I(to) density by NPY was concentration-dependent (EC(50)=10 nM). 3. In the presence of BIBP 3226, an NPY receptor antagonist that binds selectively to NPY Y1-receptors, the effect of NPY on 4-AP I(to) density was maintained. However, in the presence of BIIE 0246, a highly selective non-peptide NPY Y2-receptor antagonist, NPY was unable to increase 4-AP I(to) density. 4. The effect of NPY on 4-AP I(to) density was prevented by pretreatment with 500 ng ml( 1) pertussis toxin (PTX) and by the specific protein kinase C (PKC) inhibitor, calphostin C (100 nM). 5. Thus, short term exposure to NPY induces an increase of 4-AP I(to) density in rat ventricular myocytes mediated by Y2-receptors and involving the action of PKC via a PTX-sensitive signalling cascade. PMID- 11934811 TI - Dexrazoxane pre-treatment protects skinned rat cardiac trabeculae against delayed doxorubicin-induced impairment of crossbridge kinetics. AB - 1. Dexrazoxane (DXR, ICRF-187) has been shown both in animal studies and clinical trials to provide a substantial cardioprotection when co-administered with anthracycline drugs like Doxorubicin (DOX). In a previous study, we showed that chronic DOX treatment in rats is associated with a clear impairment of the crossbridge kinetics and shift in myosin iso-enzymes. 2. The present study was adopted to investigate whether the cardioprotective action of DXR involves preservation of the normal actin-myosin interaction. Rats were treated for 4 weeks with either DOX at a weekly dose of 2 mg kg(-1) (i.v.), or were pre injected with DXR (40 mg kg(-1), i.v.) at a 20 : 1 dose ratio 30 min prior to the DOX infusion. Rats receiving saline or DXR alone were included in the experiments. Cardiac trabeculae were isolated 4 weeks after the last infusion and were skinned with detergent. 3. Crossbridge turnover kinetics were studied after application of rapid length perturbations of varying amplitudes in Ca(2+) activated preparations. DXR treatment offered a significant protection against the DOX-induced impairment of the crossbridge kinetics in isolated cardiac trabeculae. Time constants describing transitions between different crossbridge states were restored to normal in both the quick release protocol and the slack test. DXR prevented the shift from the 'high ATPase' alpha-myosin heavy chain (MHC) isoform towards the 'low-ATPase' beta-MHC isoform in the ventricles. 4. We conclude that pre-administration of DXR in rats greatly reduces the deleterious effects of chronic DOX treatment on the trabecular actin - myosin crossbridge cycle. Preventing direct deleterious effects on the actin - myosin crossbridge system may provide a new target for preventing or reducing DOX-related cardiotoxicity and may enable patients to continue the treatment beyond currently imposed limits. PMID- 11934812 TI - Mutations inducing divergent shifts of constitutive activity reveal different modes of binding among catecholamine analogues to the beta(2)-adrenergic receptor. AB - 1. We compared the changes in binding energy generated by two mutations that shift in divergent directions the constitutive activity of the human beta(2) adrenergic receptor (beta(2)AR). 2. A constitutively activating mutant (CAM) and the double alanine replacement (AA mutant) of catechol-binding serines (S204A, S207A) in helix 5 were stably expressed in CHO cell lines, and used to measure the binding affinities of more than 40 adrenergic ligands. Moreover, the efficacy of the same group of compounds was determined as intrinsic activity for maximal adenylyl cyclase stimulation in wild-type beta(2)AR. 3. Although the two mutations had opposite effects on ligand affinity, the extents of change were in both cases largely correlated with the degree of ligand efficacy. This was particularly evident if the extra loss of binding energy due to hydrogen bond deletion in the AA mutant was taken into account. Thus the data demonstrate that there is an overall linkage between the configuration of the binding pocket and the intrinsic equilibrium between active and inactive receptor forms. 4. We also found that AA mutation-induced affinity changes for catecholamine congeners gradually lacking ethanolamine substituents were linearly correlated to the loss of affinity that such modifications of the ligand cause for wild-type receptor. This indicates that the strength of bonds between catechol ring and helix 5 is critically dependent on the rest of interactions of the beta-ethanolamine tail with other residues of the beta(2)-AR binding pocket. PMID- 11934813 TI - Acute cerebral focal ischaemia alters the adrenergic and NANC responses in the bisected rat vas deferens. AB - 1. Disturbances of the autonomic nervous system are common in right hemisphere stroke patients, including a marked decline in male sexual functions. There is a lack of information on the influence of stroke on male secondary sex organs such as the vas deferens. 2. This study investigates the effect of right brain focal ischaemia on the adrenergic and purinergic responses in isolated epididymal and prostatic portions of rat vas deferens. 3. In both epididymal and prostatic portions the concentration-response curves to noradrenaline are flattened resulting in a reduction (up to 67 - 76%) of the maximum contractile response in the tissue from ischaemic rats compared to the controls. In the prostatic portion from ischaemic rats the concentration-response curve to alpha,beta-methylene ATP was also depressed. 4. The first purinergic and the second delayed adrenergic phase to single pulse was not modified by brain ischaemia. In contrast both phasic and tonic components of the electrically induced contractions by trains of stimuli at high frequencies (2 - 30 Hz) were significantly depressed in the epididymal and prostatic portions from ischaemic rats. 5. These results demonstrate an autonomic imbalance at the level of male sexual secondary organs which may contribute to sexual impairment after stroke. PMID- 11934815 TI - Angiotensin-(1-7) is involved in the endothelium-dependent modulation of phenylephrine-induced contraction in the aorta of mRen-2 transgenic rats. AB - 1. The contribution of the local vascular production of angiotensin-(1-7) [Ang-(1 7)] to the control of alpha-adrenergic-induced contractions in the aorta of Sprague-Dawley (SD) and TGR(mRen-2)27 [mRen-2] rats was studied. 2. In mRen-2 rats, contractile responses to phenylephrine were diminished as compared to control SD rats in endothelium containing but not in endothelium-denuded vessels. L-NAME increased contractile responses to phenylephrine in mRen-2 rats and, after nitric oxide synthase blockade, responses to phenylephrine became comparable in both strains. 3. Inhibition of angiotensin-converting enzyme (ACE) by captopril potentiated contractile responses in mRen-2 rats and diminished contractile responses in SD rats, both effects being dependent on the presence of a functional endothelium. The effect of captopril in mRen-2 rats was abolished in vessels pre-incubated with Ang-(1-7). 4. Blockade of Ang-(1-7) and bradykinin (BK) receptors by A-779 and HOE 140 respectively, increased phenylephrine-induced contraction in mRen-2, but not in SD rats. This effect was seen only in endothelium-containing vessels. 5. Angiotensin II AT(1) and AT(2) receptor blockade by CV 11974 and PD 123319 did not affect the contractile responses to phenylephrine in aortas of transgenic animals but diminished the response in SD rats. This effect was only seen in the presence of a functional endothelium. 6. It is concluded that the decreased contractile responses to phenylephrine in aortas of mRen-2 rats was dependent on an intact endothelium, the local release and action of Ang-(1-7) and bradykinin. PMID- 11934814 TI - Kainate receptor-mediated apoptosis in primary cultures of cerebellar granule cells is attenuated by mitogen-activated protein and cyclin-dependent kinase inhibitors. AB - 1. Previous studies have suggested that neuronal apoptosis is the result of an abortive attempt to re-enter the cell cycle, and more recently the cyclin dependent (CDKs) and the mitogen-activated protein (MAP) kinases, two superfamilies of kinases that influence and control cell cycle progression, have been implicated in neuronal apoptosis. 2. Here, to examine whether CDK/MAPK related pathways are involved in excitotoxicity, we studied the actions of various kinase inhibitors on apoptosis induced by the ionotropic glutamate (Glu) receptor agonist, kainate (KA), in primary cultures of murine cerebellar granule cells (CGCs). 3. KA-mediated neurotoxicity was concentration-dependent, as determined by a cell viability assay monitoring the reduction of 3-(4,5 dimethylthiazole-2-yl)-2,5-diphenyltetrazolium bromide (MTT), and largely apoptotic in nature, as shown by morphological examination and labelling of DNA fragmentation in situ using terminal deoxynucleotidyl transferase (TdT)-mediated dUTP digoxigenin nick-end labelling (TUNEL). 4. KA-mediated neurotoxicity and apoptosis was completely attenuated by the mixed CDK and MAP kinase inhibitor, olomoucine, in a concentration-dependent manner (50 - 600 microM), and partially by roscovitine (1 - 100 microM), a more selective CDK inihibitor. 5. The p38 MAP kinase inhibitor, SB203580 (1 - 100 microM), partially attenuated KA receptor mediated apoptosis, as did the MAP kinase kinase inhibitors PD98509 (1 - 100 microM) and U0126 (1 - 100 microM). 6. These findings provide new evidence for a complex network of interacting pathways involving CDK/MAPK that control apoptosis downstream of KA receptor activation in excitotoxic neuronal cell death. PMID- 11934816 TI - Leukocyte rolling is exclusively mediated by P-selectinin colonic venules. AB - 1. The objective of the present study was to examine the role of the endothelial selectins (i.e. P- and E-selectin) in leukocyte-endothelium interactions in colonic venules by use of intravital microscopy. 2. Balb/c mice were exposed to dextran sodium sulphate (DSS) in the drinking water for 5 days or treated intraperitoneally (i.p.) with tumour necrosis factor-alpha (TNF-alpha) for 3 h. 3. In DSS-treated mice, mRNA of both P- and E-selectin were expressed and leukocyte rolling and adhesion was increased to 27+/-3 cells min(-1) and 36+/-8 cells mm(-1), respectively. An anti-P-selectin antibody abolished DSS-induced leukocyte rolling, whereas an antibody against E-selectin had no effect. Established leukocyte adhesion was insensitive to inhibition of the selectins. 4. DSS markedly increased production of TNF-alpha in the colon. TNF-alpha increased leukocyte rolling to 22+/-3 cells min(-1) and adhesion to 45+/-4 cells mm(-1). Only inhibition of P-selectin significantly reduced (>94%) leukocyte rolling provoked by TNF-alpha. Leukocyte adhesion was not changed by late anti-P-selectin antibody treatment. In contrast, pretreatment with the anti-P-selectin antibody not only abolished leukocyte rolling but also completely inhibited firm adhesion in response to TNF-alpha. 5. This study demonstrates that P-selectin plays an important role in leukocyte rolling in colonic venules, both in experimental colitis and when stimulated with TNF-alpha. Moreover, P-selectin-dependent leukocyte rolling was found to be a precondition for TNF-alpha-induced firm adhesion. Thus, these findings suggest that P-selectin may be a key target to reduce pathological recruitment of inflammatory cells in the colon. PMID- 11934817 TI - Alpha-1 adrenoceptor up-regulation induced by prazosin but not KMD-3213 or reserpine in rats. AB - 1. We have investigated the effects of chronic administration of prazosin (a subtype-nonspecific alpha-1 AR antagonist), KMD-3213 (an alpha-1A AR subtype specific antagonist) and reserpine (a catecholamine depletor) on the density of alpha-1 AR subtypes in various rat tissues (liver, kidney, submaxillary gland, heart and spleen). 2. Administration of prazosin (2 mg kg(-1) day(-1), i.p.) for 2 weeks did not affect K(D) values for [(3)H]-prazosin or [(3)H]-KMD-3213 of alpha-1 ARs in five rat tissues tested. However, it caused 52% up-regulation of alpha-1B AR in the spleen, and 84% and 107% up-regulation of alpha-1A- and alpha 1B ARs, respectively, in the heart. Although major subtypes of alpha-1 AR are alpha-1A AR in the submaxillary gland, alpha-1B AR in the liver, and alpha-1A and alpha-1B ARs in the kidney, these tissues showed no up-regulation. The mRNA levels of alpha-1 AR subtypes were not affected by prazosin administration in any tissue tested. 3. Neither administration of KMD-3213 (2 mg kg(-1) day(-1), i.p.) nor reserpine (0.5 - 1 mg kg(-1) day(-1), i.p.) for 2 weeks caused any change in either the binding affinity for [(3)H]-prazosin or [(3)H]-KMD-3213 or the density of the alpha-1 AR subtypes in the five rat tissues. 4. Neither prazosin nor KMD 3213 treatment reduced the noradrenaline content in the five rat tissues, in contrast to reserpine treatment, which markedly reduced it. 5. The findings of the present study demonstrated that up-regulation of alpha-1 AR is selectively caused by prazosin treatment in some tissues but neither by KMD-3213 treatment nor by chemical denervation with reserpine. These results suggest that up regulation of alpha-1 ARs is not caused by a simple blockade of sympathetic tone. PMID- 11934818 TI - Muscarinic agonist potencies at three different effector systems linked to the M(2) or M(3) receptor in longitudinal smooth muscle of guinea-pig small intestine. AB - 1. The abilities of muscarinic agonists (arecoline, bethanechol, carbachol, McN A343, methacholine, pilocarpine) to inhibit isoprenaline-induced cyclic AMP production in chopped fragments (via M(2) receptors), and to evoke cationic current (I(cat)) (via M(2) receptors) or calcium store release (via M3 receptors) in enzyme-dispersed, single voltage-clamped cells from longitudinal smooth muscle of the guinea-pig small intestine were examined. 2. All muscarinic agonists (1 - 300 microM) examined inhibited isoprenaline (1 microM)-induced accumulation of cyclic AMP, the IC(50) varying from 52 to 248 microM. However, their relative potencies to evoke this M(2) effect were not significantly correlated with their ability to evoke I(cat), also a M(2) effect, whether or not calcium stores were depleted; pilocarpine and McN-A343 inhibited the I(cat) response to carbachol. 3. Muscarinic agonists (concentration 300 or 1000 microM), except pilocarpine and McN-A343 which were ineffective, evoked Ca(2+)-activated K(+) current (I(K-Ca)) resulting from Ca(2+) store release (M(3) effect). Their effectiveness was tested by estimating residual stored calcium by subsequent application of caffeine (10 mM). The relative potencies to evoke Ca(2+) store release (M(3)) and for I(cat) activation (M(2)) were closely correlated (P<0.001). 4. These data might be explained if M(2)-mediated adenylyl cyclase inhibition and I(cat) activation involve different G proteins, or involve different populations of M(2) receptors. The observed correlation of agonist potency between I(cat) activation and Ca(2+) store release supports the proposal (Zholos & Bolton, 1997) that M(3) activation can potentiate M(2)-cationic channel coupling through Ca(2+)-independent mechanisms. PMID- 11934819 TI - Free-radical production triggered by hyperthermia contributes to heat stress induced cardioprotection in isolated rat hearts. AB - 1. Heat stress (HS) is known to protect the myocardium against ischaemic damage. It has been reported that reactive oxygen species (ROS) are abundantly produced during this stress. Since mechanisms triggering the HS-induced cardioprotection remain unknown, we investigated the role of ROS in the genesis of this protective phenomenon. 2. Rats were divided into four groups (n=8 in each group), subjected to either hyperthermia (42 degrees C internal temperature for 15 min) or sham anaesthesia and treated or not with N-2-mercaptopropionyl glycine (MPG), a synthetic antioxidant, 10 min before HS. Twenty-four hours later, their hearts were isolated, retrogradely perfused, and subjected to a 30-min occlusion of the left coronary artery followed by 120 min of reperfusion. Myocardial Hsp 27 and 70 expression was assessed by Western blot analysis (n=4). Cardiac activities of antioxidant enzymes (superoxide dismutase and glutathione peroxidase) were also examined (n=4). 3. Infarct-to-risk zone ratio was significantly reduced in HS (17+/-1.3%) compared to Sham (34.3+/-1.7%) hearts. This effect was abolished by MPG pretreatment (40.6+/-1.9% in HS+MPG vs. 39.8+/-2.5% in Sham+MPG hearts). This cardioprotection was associated with an enhanced Hsp 27 and 70 expression, which was not modified by MPG pretreatment. Antioxidant enzyme activities was not modified by heat stress or MPG pretreatment. 4. Free radical production following hyperthermia appears to play a role in the heat stress induced cardioprotection, independently of Hsp levels. Antioxidant enzyme activities do not seem to be implicated in this cardioprotective mechanism. PMID- 11934820 TI - Effects of a phosphodiesterase IV inhibitor rolipram on microsphere embolism induced defects in memory function and cerebral cyclic AMP signal transduction system in rats. AB - The effects of treatment with rolipram, a specific phosphodiesterase IV inhibitor, on learning and memory function and on the cyclic AMP/PKA/CREB signal transduction system were examined in rats with microsphere embolism (ME)-induced cerebral ischaemia. Sustained cerebral ischaemia was induced by the injection of 900 microspheres (48 microm in diameter) into the right hemisphere of the rat brain. The animals were treated once daily with 3 mg kg(-1) rolipram i.p. from 6 h after the onset of the operation for consecutive 10 days. Microsphere-embolized rats showed prolongation of the escape latency in the water maze task starting from day 7 after the operation and lasting for 3 consecutive days. Treatment with rolipram reduced the escape latency. ME decreased the cyclic AMP content, cytosolic PKA Cbeta level, and nuclear PKA Calpha and Cbeta levels, as well as reduced the pCREB level and the DNA-binding activity of CREB in the cerebral cortex and hippocampus on day 10 after the operation. These alterations were attenuated by treatment with rolipram. These results suggest that ME-induced failure in learning and memory function may be mediated by dysfunction of the cyclic AMP/PKA/CREB signal transduction system, that rolipram may ameliorate ME induced impairment of learning and memory function, and that the drug effect may be partly attributed to activation of the cyclic AMP/PKA/CREB signal transduction system. PMID- 11934821 TI - Dextran sodium sulphate-induced colitis perturbs muscarinic cholinergic control of colonic epithelial ion transport. AB - 1. Neuronal cholinergic input is an important regulator of epithelial electrolyte transport and hence water movement in the gut. 2. In this study, colitis was induced by treating mice with 4% (w v(-1)) dextran sodium-sulphate (DSS)-water for 5 days followed by 3 days of normal water. Mid-colonic segments were mounted in Ussing chambers and short-circuit current (Isc, indicates net ion movement) responses to the cholinergic agonist, carbachol (CCh; 10(-4) M)+/-tetrodotoxin, atropine (ATR), hexamethonium (HEX), naloxone or phenoxybenzamine were assessed. 3. Tissues from mice with DSS-induced colitis displayed a drop in Isc in response to CCh (-11.3+/-3.3 microA/cm(2)), while those from control mice showed a transient increase in Isc (76.3+/-13.0 microA/cm(2)). The DeltaIsc in colon from DSS-treated mice was tetrodotoxin-sensitive, atropine-insensitive and was reversed by hexamethonium (HEX+CCh=16.7+/-7.8 microA/cm(2)), indicating involvement of a nicotinic receptor. 4. CCh induced a drop in Isc in tissues from controls only when they were pretreated with the cholinergic muscarinic receptor blocker, atropine: ATR+CCh=-21.3+/-7.0 microA/cm(2). Nicotine elicited a drop in Isc in Ussing-chambered colon from both control and DSS-treated mice that was TTX sensitive. 5. The drop in Isc evoked by CCh challenge of colonic tissue from DSS treated mice or ATR+CCh challenge of control tissue was not significantly affected by blockade of opiate or alpha-adrenergic receptors by naloxone or phenoxybenzamine, respectively. 6. The data indicate that DSS-colitis reveals a nicotinic receptor that becomes important in cholinergic regulation of ion transport. PMID- 11934822 TI - Ascorbate blocks endothelium-derived hyperpolarizing factor (EDHF)-mediated vasodilatation in the bovine ciliary vascular bed and rat mesentery. AB - 1. The effects of ascorbate were assessed on vasodilatation mediated by endothelium-derived hyperpolarizing factor (EDHF) in the ciliary vascular bed of the bovine isolated perfused eye and in the rat isolated perfused mesenteric arterial bed. 2. In the bovine eye, EDHF-mediated vasodilator responses induced by acetylcholine or bradykinin were powerfully blocked when ascorbate (50 microM) was included in the perfusion medium for at least 120 min; with acetylcholine a normally-masked muscarinic vasoconstrictor response was also uncovered. 3. The blockade of EDHF-mediated vasodilatation by ascorbate was time-dependent (maximum blockade at 120 min) and concentration-dependent (10 - 150 microM). 4. Ascorbate (50 microM) also blocked acetylcholine-induced, EDHF-mediated vasodilator responses in the rat mesenteric arterial bed in a time-dependent manner (maximum blockade at 180 min). 5. The ability of ascorbate to block EDHF-mediated vasodilatation is likely to result from its reducing properties, since this action was mimicked in the bovine eye by two other reducing agents, namely, N acetyl-L-cysteine (1 mM) and dithiothreitol (100 microM), but not by the redox inactive analogue, dehydroascorbate (50 microM). 6. In conclusion, concentrations of ascorbate present in normal plasma block EDHF-mediated vasodilator responses in the bovine eye and rat mesentery. The mechanism and physiological consequences of this blockade remain to be determined. PMID- 11934823 TI - Block of Ca(2+)-channels by alpha-endosulphine inhibits insulin release. AB - 1. alpha-Endosulphine, isolated as an endogenous equivalent for sulphonylureas, is a 121-amino acids protein of 19 kDa apparent molecular mass, member of a cyclic AMP-regulated phosphoprotein family. We have previously shown that alpha endosulphine inhibits sulphonylurea binding and K(ATP) channel activity, thereby stimulating basal insulin secretion. 2. We now describe that in the perfused rat pancreas, no stimulation was detected and that alpha-endosulphine inhibited glucose stimulated insulin release. This inhibition was dose-dependent and affected both phases of insulin secretion. 3. This inhibitory effect of alpha endosulphine also occurred on MIN6 beta-cells when insulin release was stimulated either by glucose, sulphonylureas or a high K(+) depolarization. Inhibition was concentration-dependent with a half-maximal inhibition at 0.5 microM and was mirrored by inhibition of calcium influx. 4. Electrophysiological experiments demonstrated, in comparison to the effects of the sulphonylurea tolbutamide, that these inhibitory effects were linked to a direct inhibition of L-type Ca(2+) channels and were independent from a regulation of K(ATP) channels. 5. Although alpha-endosulphine is able to stimulate insulin release under specific conditions acting via modulation of K(ATP) channel activity, the present study suggests that, under physiological conditions, the peptide mainly acts to block voltage gated Ca(2+)-channels. This block leads to the inhibition of calcium influx and triggers inhibition of insulin release. 6. We conclude that alpha-endosulphine is not exclusively an endogenous equivalent for sulphonylureas and not solely a K(ATP) channel regulator. PMID- 11934824 TI - Quercetin as a novel activator of L-type Ca(2+) channels in rat tail artery smooth muscle cells. AB - 1. The aim of this study was to investigate the effects of quercetin, a natural polyphenolic flavonoid, on voltage-dependent Ca(2+) channels of smooth muscle cells freshly isolated from the rat tail artery, using either the conventional or the amphotericin B-perforated whole-cell patch-clamp method. 2. Quercetin increased L-type Ca(2+) current [I(Ca(L))] in a concentration- (pEC(50)=5.09+/ 0.05) and voltage-dependent manner and shifted the maximum of the current-voltage relationship by 10 mV in the hyperpolarizing direction, without, however, modifying the threshold and the equilibrium potential for Ca(2+). 3. Quercetin induced I(Ca(L)) stimulation was reversible upon wash-out. T-type Ca(2+) current was not affected by quercetin. Quercetin shifted the voltage dependence of the steady-state inactivation and activation curves to more negative potentials by about 5.5 and 7.5 mV respectively, in the mid-potential of the curves as well as increasing the slope of activation. Quercetin slowed both the activation and the deactivation kinetics of the I(Ca(L)). The inactivation time course was also slowed but only at voltages higher than 10 mV. Moreover quercetin slowed the rate of recovery from inactivation. 4. These results prove quercetin to be a naturally occurring L-type Ca(2+) channel activator. PMID- 11934825 TI - Pharmacological characterization of F-180: a selective human V(1a) vasopressin receptor agonist of high affinity. AB - 1. The pharmacological properties of F-180, a vasopressin (VP) structural analogue, were determined on CHO cells expressing the different human vasopressin and oxytocin (OT) receptor subtypes. Binding experiments revealed that F-180 exhibited a high affinity for the human V(1a) receptor subtype (K(i)=11 nM) and was selective for this receptor subtype. 2. Functional studies performed on CHO cells expressing human V(1a) receptors indicate that similarly to AVP, F-180 can stimulate the accumulation of inositol phosphate. The activation constant (K(act)) for both F-180 and AVP was 1.7 nM. F-180 was also an agonist for the human V(2) and V(1b) receptor subtypes and an antagonist for the human OT receptor. 3. Since marked species pharmacological differences for vasopressin receptors have been described, we studied the properties of F-180 on various mammalian species. F-180 showed high affinity and good selectivity for human and bovine V(1a) receptors, but weak affinity and non selective properties for rat V(1a) receptors. 4. To assess the functional properties of F-180 on a native biological model, we performed studies on primary cultures of cells from bovine zona fasciculata (ZF). As AVP, F-180 stimulated inositol phosphate accumulation and cortisol secretion with similar efficiency. 5. In conclusion, we demonstrate that F-180 is the first selective V(1a) agonist described for human and bovine vasopressin receptors. Therefore F-180 can be used as a powerful pharmacological tool to characterize the actions of vasopressin that are mediated by V(1a) receptor subtypes. PMID- 11934826 TI - Adrenomedullin: an autocrine/paracrine factor for cardiorenal protection. PMID- 11934827 TI - Smooth muscle-specific transcription without a CArG box element. PMID- 11934828 TI - IGF-1 Overexpression rescues the failing heart. PMID- 11934829 TI - Evidence for cardiomyocyte repopulation by extracardiac progenitors in transplanted human hearts. AB - Human myocardium has long been considered to have essentially no intrinsic regenerative capacity. Recent studies in rodent models, however, have suggested the presence of an extracardiac stem cell population, perhaps in bone marrow, that is capable of some reconstitution of cardiomyocytes after injury. To determine whether similar mechanisms exist in the human heart, we evaluated human female allograft hearts transplanted into male patients. The presence of Y chromosomes in cardiomyocytes would indicate these cells arose from the recipient, rather than the donor heart. We identified 5 male patients who had retained a female heart at least 9 months before death and necropsy. Remarkably, in each case, the transplanted heart contained a minute but readily detectable fraction of Y chromosome-positive cardiomyocytes. The mean percentage of cardiomyocytes arising from the host was estimated to be 0.04% with a median of 0.016%. Most Y-positive cardiomyocytes were associated with regions of acute rejection, suggesting such chimerism involves an injury event. Furthermore, the sole patient whose immediate cause of death was allograft rejection showed a much higher percentage of host-derived cardiomyocytes, up to 29% in local, 1-mm(2) "hot spots." Thus, adult humans have extracardiac progenitor cells capable of migrating to and repopulating damaged myocardium, but this process occurs at very low levels. PMID- 11934830 TI - Cardiac-specific IGF-1 expression attenuates dilated cardiomyopathy in tropomodulin-overexpressing transgenic mice. AB - To test the hypothesis that early interventional treatment with insulin-like growth factor-1 (IGF-1) alleviates subsequent development of dilated cardiomyopathy, cardiac-specific IGF-1 expression was introduced by selective cross-breeding into a transgenic mouse model of heart failure that displays phenotypic characteristics of severe dilation. Hemodynamic, structural, and cellular parameters of the heart were compared between nontransgenic, tropomodulin-overexpressing cardiomyopathic, and the hybrid tropomodulin/IGF-1 overexpressing mice. Beneficial effects of IGF-1 were apparent by multiple indices of cardiac structure and function, including normalization of heart mass, anatomy, hemodynamics, and apoptosis. IGF-1 expression also acted as a proliferative stimulus as evidenced by calculated increases in myocyte number as well as expression of Ki67, a nuclear marker of cellular replication. Cellular analyses revealed that IGF-1 inhibited characteristic cardiomyocyte elongation in dilated hearts and restored calcium dynamics comparable to that observed in normal cells. Collectively, these results provide novel information regarding the ability of IGF-1 to inhibit progression of cardiomyopathic disease in a defined model system and suggest that heart failure may benefit from early interventional IGF-1 treatment. PMID- 11934831 TI - Phosphorylation of troponin I controls cardiac twitch dynamics: evidence from phosphorylation site mutants expressed on a troponin I-null background in mice. AB - The cardiac myofilament protein troponin I (cTnI) is phosphorylated by protein kinase C (PKC), a family of serine/threonine kinases activated within heart muscle by a variety of agonists. cTnI is also a substrate for cAMP-dependent protein kinase (PKA) activated during beta-adrenergic signaling. To investigate the role of cTnI phosphorylation in contractile regulation by these pathways, we generated transgenic mice harboring a mutated cTnI protein lacking phosphorylation sites for PKC (serine(43/45) and threonine(144) mutated to alanine) and for PKA (serine(23/24) mutated to alanine). Transgenic mice were interbred with cTnI-knockout mice to ensure the absence of endogenous phosphorylatable cTnI. Here, we report that regulation of myocyte twitch kinetics by beta-stimulation and by endothelin-1 was altered in myocytes containing mutant cTnI. In wild-type myocytes, the beta-agonist isoproterenol decreased twitch duration and relaxation time constant (tau) by 37% to 44%. These lusitropic effects of isoproterenol were reduced by about half in nonphosphorylatable cTnI mutant myocytes and were absent in cTnI mutants also lacking phospholamban (generated by crossing cTnI mutants with phospholamban-knockout mice). These observations are consistent with important roles for both cTnI and phospholamban phosphorylation in accelerating relaxation after beta-adrenergic stimulation. In contrast, endothelin-1 increased twitch duration by 32% and increased tau by 58%. These endothelin-1 effects were substantially blunted in nonphosphorylatable cTnI myocytes, indicating that PKC phosphorylation of cTnI slows cardiac relaxation and increases twitch duration. We propose that beta-agonists and endothelin-1 regulate cardiac twitch dynamics in opposite directions in part through phosphorylation of the myofilament protein cTnI on distinct sites. PMID- 11934833 TI - Optical mapping of arrhythmias induced by strong electrical shocks in myocyte cultures. AB - Strong electrical shocks can induce arrhythmias, which might explain why shocks fail to defibrillate. In this work, the localization of arrhythmia source and the relationship with local changes of transmembrane potential (V(m)) were determined in geometrically defined cell cultures using optical mapping technique. Uniform field shocks with strength (E) of 10 to 50 V/cm were applied across cell strands with width of 0.2 and 0.8 mm. The threshold for arrhythmia induction was dependent on the strand width: in the 0.8- and 0.2-mm strands, arrhythmias were induced at E>/=20.6+/-1.8 V/cm (n=8) and E>/=30.3+/-1.8 V/cm (n=8), respectively. At the same shock strength, the arrhythmia rate and duration were larger in the wider strands. During shocks that induced arrhythmias, the V(m) waveforms on the anodal side revealed a positive V(m) shift that followed the initial large hyperpolarization and postshock elevation of the diastolic V(m). These V(m) changes were absent during failed shocks. To determine the localization of the arrhythmia source, arrhythmias were induced in narrow cell strands containing regions of local expansion. Optical mapping of the first extrabeat with a coupling interval of 315+/-60 ms revealed that in the majority of cases (9 out of 13) the source of arrhythmias was localized in the areas of shock-induced hyperpolarization. Thus, (1) induction of postshock arrhythmias, their rate, and their duration strongly depend on the tissue structure; (2) arrhythmia induction coincides with the appearance of a positive V(m) shift in the areas of hyperpolarization; and (3) the source of postshock arrhythmias is located in the areas of shock-induced hyperpolarization. PMID- 11934832 TI - Role of endogenous adrenomedullin in the regulation of vascular tone and ischemic renal injury: studies on transgenic/knockout mice of adrenomedullin gene. AB - Adrenomedullin (AM) is a potent depressor peptide whose vascular action is suggested to involve nitric oxide (NO) release. To explore the role of endogenous AM in vascular and renal function, we examined the effects of acetylcholine (ACh), AM, and AM receptor antagonists AM(22-52) and CGRP(8-37) on the renal perfusion pressure (RPP) of kidneys isolated from AM transgenic (TG)/heterozygote knockout (KO) mice and wild-type littermates (WT). Furthermore, we evaluated the renal function and histology 24 hours after bilateral renal artery clamp for 45 minutes in TG, KO, and WT mice. Baseline RPP was significantly lower in TG than in KO and WT mice (KO 93.4+/-4.6, WT 85.8+/-4.2, TG 72.4+/-2.4 mm Hg [mean+/-SE], P<0.01). ACh and AM caused a dose-related reduction in RPP, but the degree of vasodilatation was smaller in TG than that in KO and WT (%DeltaRPP 10(-7) mol/L ACh: KO -48.1+/-3.9%, WT -57.5+/-5.6%, TG -22.8+/-4.8%, P<0.01), whereas N(G) nitro-L-arginine methyl ester (L-NAME) caused greater vasoconstriction in TG (%DeltaRPP 10(-4) mol/L: KO 33.1+/-3.3%, WT 55.5+/-7.2%, TG 152.6+/-21.2%, P<0.01). Both AM antagonists increased RPP in TG to a greater extent compared with KO and WT mice (%DeltaRPP 10(-6) mol/L CGRP(8-37): KO 12.8+/-2.6%, WT 19.4+/ 3.6%, TG 41.8+/-8.7%, P<0.01). In mice with ischemic kidneys, serum levels of urea nitrogen and renal damage scores showed smaller values in TG and greater values in KO mice (urea nitrogen: KO 104+/-5>WT 98+/-15>TG 38+/-7 mg/dL, P<0.05 each). Renal NO synthase activity was also greater in TG mice. However, the differences in serum urea nitrogen and renal damage scores among the 3 groups of mice were not observed in mice pretreated with L-NAME. In conclusion, AM antagonists increased renal vascular tone in WT as well as in TG, suggesting that endogenous AM plays a role in the physiological regulation of the vascular tone. AM is likely to protect renal tissues from ischemia/reperfusion injury through its NO releasing activity. PMID- 11934834 TI - Autocrine regulation of myocyte Cx43 expression by VEGF. AB - Cardiac myocytes can rapidly adjust their expression of gap junction channel proteins in response to changes in load. Previously, we showed that after only 1 hour of linear pulsatile stretch (110% of resting cell length; 3 Hz), expression of connexin43 (Cx43) by cultured neonatal rat ventricular myocytes is increased by approximately 2-fold and impulse propagation is significantly more rapid. In the present study, we tested the hypothesis that vascular endothelial growth factor (VEGF), acting downstream of transforming growth factor-beta (TGF-beta), mediates stretch-induced upregulation of Cx43 expression by cardiac myocytes. Incubation of nonstretched cells with exogenous VEGF (100 ng/mL) or TGF-beta (10 ng/mL) for 1 hour increased Cx43 expression by approximately 1.8-fold, comparable to that observed in cells subjected to pulsatile stretch for 1 hour. Stretch induced upregulation of Cx43 expression was blocked by either anti-VEGF antibody or anti-TGF-beta antibody. Stretch-induced enhancement of conduction was also blocked by anti-VEGF antibody. ELISA assay showed that VEGF was secreted into the culture medium during stretch. Furthermore, stretch-conditioned medium stimulated Cx43 expression in nonstretched cells. This effect was also blocked by anti-VEGF antibody. Upregulation of Cx43 expression stimulated by exogenous TGF-beta was blocked by anti-VEGF antibody, but VEGF-stimulation of Cx43 expression was not blocked by anti-TGF-beta antibody. Thus, stretch-induced upregulation of Cx43 expression is mediated, at least in part, by VEGF, which acts downstream of TGF beta. Because the cultures contained only approximately 5% nonmyocytic cells, these results indicate that myocyte-derived VEGF, secreted in response to stretch, acts in an autocrine fashion to enhance intercellular coupling. PMID- 11934835 TI - Nucleotide receptors involved in UTP-induced rat arterial smooth muscle cell migration. AB - Many factors have been shown to be involved in the development of hyperplasic lesions of vessels, but the role of extracellular nucleotides remains largely unknown. The presence of P2Y and P2X nucleotide receptors on arterial endothelial and smooth muscle cells suggests a potential role for nucleotides in the vessel pathophysiology. Although the role of P2X in physiology of vessels is well documented, that of P2Y is not completely understood. We recently demonstrated that extracellular nucleotides, and particularly UTP, induced migration of cultured arterial smooth muscle cells (ASMCs). This migration is dependent on osteopontin expression and involves the Rho and mitogen-activated protein (MAP) kinase pathways. An important question is to determine the specific role of the different P2Y receptors of rat ASMCs in the UTP-induced migration process. Therefore, we first quantified mRNA levels of P2Y(2), P2Y(4), and P2Y(6) nucleotide receptors in cultured rat ASMCs by a competitive RT-PCR approach and demonstrated that P2Y(2) is the most highly expressed among these receptors potentially involved in the UTP-mediated response. In addition to UTP, UDP also induced ASMC migration even when UTP regeneration was inhibited, suggesting the involvement of UDP receptor P2Y(6). Moreover, suramin, a specific antagonist of rat P2Y(2) receptor, acted as an inhibitor of UTP-induced migration. Taken together, these results suggest a prominent role for the UTP receptor, P2Y(2), and for the UDP receptor, P2Y(6), in UTP-induced rat ASMC migration. PMID- 11934836 TI - Regulation of eNOS expression in brain endothelial cells by perinuclear EP(3) receptors. AB - We reported upregulation of endothelial nitric oxide synthase (eNOS) by PGE(2) in tissues and presence of perinuclear PGE(2) receptors (EP). We presently studied mechanisms by which PGE(2) induces eNOS expression in cerebral microvessel endothelial cells (ECs). 16,16-Dimethyl PGE(2) and selective EP(3) receptor agonist M&B28767 increased eNOS expression in ECs and the NO-dependent vasorelaxant responses induced by substance P on cerebral microvessels. These effects could be prevented by prostaglandin transporter blocker bromcresol green and actinomycin D. EP(3) immunoreactivity was confirmed on plasma and perinuclear membrane of ECs. M&B28767 increased eNOS RNA expression in EC nuclei, and this effect was augmented by overexpression of EP(3) receptors. M&B28767 also induced increased phosphorylation of Erk-1/2 and Akt, as well as changes in membrane potential revealed by the potentiometric fluorescent dye RH421, which were prevented by iberiotoxin; perinuclear K(Ca) channels were detected, and their functionality corroborated by NS1619-induced Ca(2+) signals and nuclear membrane potential changes. Moreover, pertussis toxin, Ca(2+) chelator, and channel blockers EGTA, BAPTA, and SK&F96365, as well as K(Ca) channel blocker iberiotoxin, protein-kinase inhibitors wortmannin and PD 98059, and NF-kappaB inhibitor pyrrolidine dithiocarbamate prevented M&B28767-induced increase in Ca(2+) transients and/or eNOS expression in EC nuclei. We describe for the first time that PGE(2) through its access into cell by prostaglandin transporters induces eNOS expression by activating perinuclear EP(3) receptors coupled to pertussis toxin-sensitive G proteins, a process that depends on nuclear envelope K(Ca) channels, protein kinases, and NF-kappaB; the roles for nuclear EP(3) receptors seem different from those on plasma membrane. PMID- 11934837 TI - Identification of IEX-1 as a biomechanically controlled nuclear factor-kappaB target gene that inhibits cardiomyocyte hypertrophy. AB - Biomechanical strain is a stimulus for cardiomyocyte hypertrophy and heart failure, but the underlying molecular mechanisms remain incompletely understood. Using an in vivo murine model of pressure overload and an in vitro model of mechanical stimulation of primary cardiomyocytes, we identified iex-1 as a gene activated during the early response of cardiomyocytes to hypertrophic stimuli and as a gene product that inhibits hypertrophy without affecting cardiomyocyte viability. On stimulation of cardiomyocytes, iex-1 mRNA and protein expression increased and translocation of the gene product to the cardiomyocyte nucleus occurred. iex-1 has previously been proposed as a mediator of NF-kappaB-dependent cell survival and growth in tumor cells. Here, we demonstrate that the biomechanical induction of iex-1 in cardiomyocytes was NF-kappaB-dependent, as overexpression of the NF-kappaB inhibitor IkappaBalpha completely inhibited strain-mediated iex-1 mRNA accumulation. The functional role of iex-1 was investigated by overexpressing wild-type iex-1 with replication-defective adenoviral gene transfer. Overexpression of iex-1 abolished cardiomyocyte hypertrophy by mechanical strain, phenylephrine, or endothelin-1 at levels that did not affect cell viability. These studies identify iex-1 as a biomechanical stress-inducible and NF-kappaB-dependent gene in cardiac muscle cells during the acute phase of hypertrophy with negative growth regulatory effects that may counterbalance early hypertrophic responses in activated cardiomyocytes. PMID- 11934838 TI - A dominant-negative p65 PAK peptide inhibits angiogenesis. AB - PAK1 is a protein kinase downstream of the small GTPases Rac and Cdc42 that previous work has implicated in endothelial cell migration via modulation of cell contraction. The first proline-rich region of PAK that binds to an SH3 domain from the adapter protein NCK was responsible for these dominant-negative effects. To test the role of PAK in angiogenesis, we prepared a peptide in which the proline-rich region was fused to the polybasic sequence from the HIV Tat protein to facilitate entry into cells. We show that the short peptide selectively binds NCK, whereas a mutant peptide does not. Treatment of cells with the PAK peptide but not the control peptide disrupts localization of PAK. This peptide specifically inhibited endothelial cell migration and contractility similarly to full-length dominant-negative PAK. In an in vitro tube-forming assay, the PAK peptide specifically blocked formation of multicellular networks. In an in vivo chick chorioallantoic membrane assay, the PAK peptide specifically blocked angiogenesis. These results, therefore, suggest a role for PAK in angiogenesis. PMID- 11934839 TI - PPAR activators as antiinflammatory mediators in human T lymphocytes: implications for atherosclerosis and transplantation-associated arteriosclerosis. AB - Activation of T lymphocytes and their ensuing elaboration of proinflammatory cytokines, such as interferon (IFN)-gamma, represent a critical step in atherogenesis and arteriosclerosis. IFNgamma pathways also appear integral to the development of transplantation-associated arteriosclerosis (Tx-AA), limiting long term cardiac allograft survival. Although disruption of these IFNgamma signaling pathways limits atherosclerosis and Tx-AA in animals, little is known about inhibitory regulation of proinflammatory cytokine production in humans. The present study investigated whether activators of peroxisome proliferator activated receptor (PPAR)alpha and PPARgamma, with their known antiinflammatory effects, might regulate the expression of proinflammatory cytokines in human CD4 positive T cells. Isolated human CD4-positive T cells express PPARalpha and PPARgamma mRNA and protein. Activation of CD4-positive T cells by anti-CD3 monoclonal antibodies significantly increased IFNgamma protein secretion from 0 to 504+/-168 pg/mL, as determined by ELISA. Pretreatment of cells with well established PPARalpha (WY14643 or fenofibrate) or PPARgamma (BRL49653/rosiglitazone or pioglitazone) activators reduced anti-CD3-induced IFNgamma secretion in a concentration-dependent manner. PPAR activators also inhibited TNFalpha and interleukin-2 protein expression. In addition, PPAR activators markedly reduced cytokine mRNA expression in these cells. Such antiinflammatory actions were also evident in cell-cell interactions with medium conditioned by PPAR activator-treated T cells attenuating human monocyte CD64 expression and human endothelial cell major histocompatibility complex class II induction. Thus, activation of PPARalpha and PPARgamma in human CD4-positive T cells limits the expression of proinflammatory cytokines, such as IFNgamma, yielding potential therapeutic benefits in pathological processes, such as atherosclerosis and Tx-AA. PMID- 11934840 TI - Leptin induces endothelin-1 in endothelial cells in vitro. AB - Leptin, a protein encoded by the obese gene, is produced by adipocytes and released into the bloodstream. In obese humans, serum leptin levels are increased and correlate with the individual's body mass index and blood pressure. Elevated serum concentrations of endothelin-1 (ET-1), a potent vasoconstrictor and mitogen, were also observed in obese subjects. The pathomechanisms underlying this ET-1 increase in obesity are poorly understood. In the present study, we investigated the influence of the ob gene product leptin on the expression of ET 1 in human umbilical vein endothelial cells (HUVECs). Binding studies using (125)I-radiolabeled leptin revealed high- and low-affinity leptin binding sites on HUVECs (Kd1=13.1+/-3.1 nmol/L and Kd2=1390+/-198 nmol/L, respectively), mediating a time- and dose-dependent increase of ET-1 mRNA expression and protein secretion after incubation of HUVECs with leptin. This leptin-induced ET-1 expression was inhibited by preincubation of HUVECs with 0.75 micromol/L antisense phosphorothioate oligonucleotides directed against the leptin receptor Ob-Rb. Furthermore, after incubation with leptin, increased nuclear staining of c fos and c-jun, the major components of the transcription factor AP-1, and increased AP-1 DNA binding were observed. Transient transfection studies with ET 1 promoter constructs showed that leptin-induced promoter activity was abolished in the absence of AP-1 binding sites or by cotransfection with a plasmid overexpressing a mutated jun, which is able to bind c-fos but not DNA. Thus, leptin upregulates ET-1 production in HUVECs via a mechanism potentially involving jun binding members of the bZIP family. PMID- 11934841 TI - Aspirin prevents Escherichia coli lipopolysaccharide- and Staphylococcus aureus induced downregulation of endothelial nitric oxide synthase expression in guinea pig pericardial tissue. AB - The aim was to analyze whether pericardial tissue expresses endothelial NO synthase (eNOS) protein and to determine the presence of cytosolic proteins that bind to eNOS mRNA. The effect of aspirin on the above-mentioned parameters was also analyzed. eNOS protein was expressed in pericardial tissue from male guinea pigs. Escherichia coli lipopolysaccharide (LPS, 10 microgram/mL) and Staphylococcus aureus endotoxin (SA, 10 microgram/mL) reduced eNOS protein expression and shortened the half-life of the eNOS messenger. Under basal conditions, cytosolic extracts from pericardial samples bound to the 3' untranslated region (3'-UTR) of eNOS mRNA, which was enhanced by LPS and SA. Proteinase K fully prevented the binding of cytosolic pericardial extracts to 3' UTR of eNOS mRNA, suggesting the involvement of proteins that were further characterized as 60- and 51-kDa proteins. Aspirin (1 to 10 mmol/L) restored eNOS expression in either LPS- and SA-stimulated pericardial samples and reduced the binding activity of the pericardial cytosolic proteins to 3'-UTR of eNOS mRNA. Indomethacin also reduced the downregulation of eNOS by LPS and diminished the binding activity of the cytosolic proteins, although higher doses of indomethacin than of aspirin were needed to improve these parameters. In conclusion, eNOS protein is expressed in guinea pig pericardial tissue. LPS and SA stimulate the binding activity of pericardial cytosolic proteins to 3'-UTR of eNOS mRNA and reduce eNOS protein expression. High doses of aspirin and indomethacin protect eNOS protein expression and reduce the binding activity of the cytosolic proteins to 3'-UTR of eNOS mRNA, suggesting an inverse association between the presence of these cytosolic proteins and eNOS expression. PMID- 11934842 TI - Characterization of the mouse aortic carboxypeptidase-like protein promoter reveals activity in differentiated and dedifferentiated vascular smooth muscle cells. AB - The dedifferentiation and proliferation of vascular smooth muscle cells (VSMCs) contribute to the formation of vascular lesions. In this study, the regulation of aortic carboxypeptidase-like protein (ACLP) expression in VSMCs was investigated. After mouse carotid injury, the expression of ACLP increases in the dedifferentiated VSMCs of the neointima in a pattern that differs from that of smooth muscle alpha-actin. To better understand the regulation of ACLP in VSMCs, we characterized the 21-exon mouse ACLP gene and 5'-flanking region and examined its promoter activity. In transient transfection assays, 2.5 kb of the ACLP 5' flanking sequence directed high levels of luciferase reporter activity in primary cultured rat aortic smooth muscle cells, and this activity was not dependent on serum response factor. We identified a positive element between base pairs -156 and -122 by analysis of 5' deletion and mutant constructs. By use of electrophoretic mobility shift assays with rat aortic smooth muscle cell nuclear extracts, Sp1 and Sp3 transcription factors bound to this region, and transfection assays in D.Mel.2 cells revealed that both Sp1 and Sp3 transactivated the ACLP promoter. Transgenic mice harboring the -2.5-kb ACLP promoter upstream from a nuclear-targeted LacZ gene were generated, and expression was detected in the VSMCs of large blood vessels, arterioles, and veins. Interestingly, ACLP promoter-LacZ reporter activity increased within the neointimal VSMCs of injured carotid vessels, consistent with the expression of the endogenous ACLP protein. The ACLP promoter may provide a novel tool to target gene expression to dedifferentiated VSMCs. PMID- 11934843 TI - Double-edged role of statins in angiogenesis signaling. AB - 3-Hydroxy-3-methylglutaryl coenzyme A reductase inhibitors (statins) exert potent vasculoprotective effects. However, the potential contribution to angiogenesis is controversial. In the present study, we demonstrate that atorvastatin dose dependently affects endothelial cell migration and angiogenesis. In vivo relevant concentrations of 0.01 to 0.1 micromol/L atorvastatin or mevastatin promote the migration of mature endothelial cells and tube formation. Moreover, atorvastatin also increases migration and the potency to form vessel structures of circulating endothelial progenitor cells, which may contribute to vasculogenesis. In contrast, higher concentrations (>0.1 micromol/L atorvastatin) block angiogenesis and migration by inducing endothelial cell apoptosis. The dose-dependent promigratory and proangiogenic effects of atorvastatin on mature endothelial cells are correlated with the activation of the phosphatidylinositol 3-kinase-Akt pathway, as determined by the phosphorylation of Akt and endothelial NO synthase (eNOS) at Ser1177. In addition, the stimulation of migration and tube formation was blocked by phosphatidylinositol 3-kinase inhibitors. In contrast, the well established stabilization of eNOS mRNA was achieved only at higher concentrations, suggesting that posttranscriptional activation rather than an increase in eNOS expression mediates the proangiogenic effect of atorvastatin. Taken together, these data suggest that statins exert a double-edged role in angiogenesis signaling by promoting the migration of mature endothelial cells and endothelial progenitor cells at low concentrations, whereas the antiangiogenic effects were achieved only at high concentrations. PMID- 11934844 TI - Different signaling pathways induce apoptosis in endothelial cells and cardiac myocytes during ischemia/reperfusion injury. AB - Apoptosis contributes, with necrosis, to the cardiac cell loss after ischemia/reperfusion injury. The apoptotic cascade is initiated either by mitochondrial damage and activation of caspase-9 or by death receptor ligation and activation of caspase-8. In the present study, performed in the isolated rat heart exposed either to ischemia alone or ischemia followed by reperfusion, cleavage of caspase-9 was observed primarily in endothelial cells. Conversely, caspase-8 cleavage was only found in cardiomyocytes, where it progressively increased throughout reperfusion. Addition of a specific caspase-9 inhibitor to the perfusate before ischemia prevented endothelial apoptosis, whereas preischemic infusion of a specific caspase-8 inhibitor affected only myocyte apoptosis. Additionally, caspase-8-mediated BID processing was observed only during reperfusion. Production of tBID then sustains mitochondrial injury and perpetuates caspase-9 activation. PMID- 11934845 TI - beta-modulation of pacemaker rate: novel mechanism or novel mechanics of an old one? PMID- 11934846 TI - Methodological considerations on the detection of 3-nitrotyrosine in the cardiovascular system. PMID- 11934847 TI - Erasing genomic imprinting memory in mouse clone embryos produced from day 11.5 primordial germ cells. AB - Genomic imprinting is an epigenetic mechanism that causes functional differences between paternal and maternal genomes, and plays an essential role in mammalian development. Stage-specific changes in the DNA methylation patterns of imprinted genes suggest that their imprints are erased some time during the primordial germ cell (PGC) stage, before their gametic patterns are re-established during gametogenesis according to the sex of individuals. To define the exact timing and pattern of the erasure process, we have analyzed parental-origin-specific expression of imprinted genes and DNA methylation patterns of differentially methylated regions (DMRs) in embryos, each derived from a single day 11.5 to day 13.5 PGC by nuclear transfer. Cloned embryos produced from day 12.5 to day 13.5 PGCs showed growth retardation and early embryonic lethality around day 9.5. Imprinted genes lost their parental-origin-specific expression patterns completely and became biallelic or silenced. We confirmed that clones derived from both male and female PGCs gave the same result, demonstrating the existence of a common default state of genomic imprinting to male and female germlines. When we produced clone embryos from day 11.5 PGCs, their development was significantly improved, allowing them to survive until at least the day 11.5 embryonic stage. Interestingly, several intermediate states of genomic imprinting between somatic cell states and the default states were seen in these embryos. Loss of the monoallelic expression of imprinted genes proceeded in a step-wise manner coordinated specifically for each imprinted gene. DNA demethylation of the DMRs of the imprinted genes in exact accordance with the loss of their imprinted monoallelic expression was also observed. Analysis of DNA methylation in day 10.5 to day 12.5 PGCs demonstrated that PGC clones represented the DNA methylation status of donor PGCs well. These findings provide strong evidence that the erasure process of genomic imprinting memory proceeds in the day 10.5 to day 11.5 PGCs, with the timing precisely controlled for each imprinted gene. The nuclear transfer technique enabled us to analyze the imprinting status of each PGC and clearly demonstrated a close relationship between expression and DNA methylation patterns and the ability of imprinted genes to support development. PMID- 11934848 TI - The onecut transcription factor HNF6 is required for normal development of the biliary tract. AB - During liver development, hepatoblasts differentiate into hepatocytes or biliary epithelial cells (BEC). The BEC delineate the intrahepatic and extrahepatic bile ducts, and the gallbladder. The transcription factors that control the development of the biliary tract are unknown. Previous work has shown that the onecut transcription factor HNF6 is expressed in hepatoblasts and in the gallbladder primordium. We now show that HNF6 is also expressed in the BEC of the developing intrahepatic bile ducts, and investigate its involvement in biliary tract development by analyzing the phenotype of Hnf6(-/-) mice. In these mice, the gallbladder was absent, the extrahepatic bile ducts were abnormal and the development of the intrahepatic bile ducts was perturbed in the prenatal period. The morphology of the intrahepatic bile ducts was identical to that seen in mice whose Hnf1beta gene has been conditionally inactivated in the liver. HNF1beta expression was downregulated in the intrahepatic bile ducts of Hnf6(-/-) mice during development. Furthermore, we found that HNF6 can stimulate the Hnf1beta promoter. We conclude that HNF6 is essential for differentiation and morphogenesis of the biliary tract and that intrahepatic bile duct development is controlled by a HNF6-->HNF1beta cascade. PMID- 11934849 TI - Bile system morphogenesis defects and liver dysfunction upon targeted deletion of HNF1beta. AB - The inactivation of the Hnf1beta gene identified an essential role in epithelial differentiation of the visceral endoderm and resulted in early embryonic death. In the present study, we have specifically inactivated this gene in hepatocytes and bile duct cells using the Cre/loxP system. Mutant animals exhibited severe jaundice caused by abnormalities of the gallbladder and intrahepatic bile ducts (IHBD). The paucity of small IHBD was linked to a failure in the organization of duct structures during liver organogenesis, suggesting an essential function of Hnf1b in bile duct morphogenesis. Mutant mice also lacked interlobular arteries. As HNF1beta is not expressed in these cells, it further emphasizes the link between arterial and biliary formation. Hepatocyte metabolism was also affected and we identified hepatocyte-specific HNF1beta target genes involved in bile acids sensing and in fatty acid oxidation. PMID- 11934850 TI - A positive role for patched-smoothened signaling in promoting cell proliferation during normal head development in Drosophila. AB - The transmembrane receptor Patched regulates several developmental processes in both invertebrates and vertebrates. In vertebrates, Patched also acts as a tumor suppressor. The Patched pathway normally operates by negatively regulating Smoothened, a G-protein-coupled receptor; binding of Hedgehog ligand to Patched relieves this negative interaction and allows signaling by Smoothened. We show that Ptc regulates Drosophila head development by promoting cell proliferation in the eye-antennal disc. During head morphogenesis, Patched positively interacts with Smoothened, which leads to the activation of Activin type I receptor Baboon and stimulation of cell proliferation in the eye-antennal disc. Thus, loss of Ptc or Smoothened activity affects cell proliferation in the eye-antennal disc and results in adult head capsule defects. Similarly, reducing the dose of smoothened in a patched background enhances the head defects. Consistent with these results, gain-of-function Hedgehog interferes with the activation of Baboon by Patched and Smoothened, leading to a similar head capsule defect. Expression of an activated form of Baboon in the patched domain in a patched mutant background completely rescues the head defects. These results provide insight into head morphogenesis, a process we know very little about, and reveal an unexpected non-canonical positive signaling pathway in which Patched and Smoothened function to promote cell proliferation as opposed to repressing it. PMID- 11934851 TI - Reciprocal functions of the Drosophila yellow and ebony proteins in the development and evolution of pigment patterns. AB - Body coloration affects how animals interact with the environment. In insects, the rapid evolution of black and brown melanin patterns suggests that these are adaptive traits. The developmental and molecular mechanisms that generate these pigment patterns are largely unknown. We demonstrate that the regulation and function of the yellow and ebony genes in Drosophila melanogaster play crucial roles in this process. The Yellow protein is required to produce black melanin, and is expressed in a pattern that correlates with the distribution of this pigment. Conversely, Ebony is required to suppress some melanin formation, and is expressed in cells that will produce both melanized and non-melanized cuticle. Ectopic expression of Ebony inhibits melanin formation, but increasing Yellow expression can overcome this effect. In addition, ectopic expression of Yellow is sufficient to induce melanin formation, but only in the absence of Ebony. These results suggest that the patterns and levels of Yellow and Ebony expression together determine the pattern and intensity of melanization. Based on their functions in Drosophila melanogaster, we propose that changes in the expression of Yellow and/or Ebony may have evolved with melanin patterns. Consistent with our hypothesis, we find that Yellow and Ebony are expressed in complementary spatial patterns that correlate with the formation of an evolutionary novel, male specific pigment pattern in Drosophila biarmipes wings. These findings provide a developmental and genetic framework for understanding the evolution of melanin patterns. PMID- 11934852 TI - Xcl1 causes delayed oblique periclinal cell divisions in developing maize leaves, leading to cellular differentiation by lineage instead of position. AB - Differentiation of plant cells is regulated by position-dependent mechanisms rather than lineage. The maize Extra cell layers1 (Xcl1) mutation causes oblique, periclinal divisions to occur in the protoderm layer. These protodermal periclinal divisions occur at the expense of normal anticlinal divisions and cause the production of extra cell layers with epidermal characteristics, indicating that cells are differentiating according to lineage instead of position. Mutant kernels have several aleurone layers instead of one, indicating that Xcl1 alters cell division orientation in cells that divide predominantly in the anticlinal plane. Dosage analysis of Xcl1 reveals that the mutant phenotype is caused by overproduction of a normal gene product. This allows cells that have already received differentiation signals to continue to divide in aberrant planes and suggests that the timing of cell division determines differentiation. Cells that divide early and in the absence of differentiation signals use positional information, while cells that divide late after perceiving differentiation signals use lineage information instead of position. PMID- 11934853 TI - Mash1 and Ngn1 control distinct steps of determination and differentiation in the olfactory sensory neuron lineage. AB - bHLH transcription factors are expressed sequentially during the development of neural lineages, suggesting that they operate in genetic cascades. In the olfactory epithelium, the proneural genes Mash1 and neurogenin1 are expressed at distinct steps in the same olfactory sensory neuron lineage. Here, we show by loss-of-function analysis that both genes are required for the generation of olfactory sensory neurons. However, their mutant phenotypes are strikingly different, indicating that they have divergent functions. In Mash1 null mutant mice, olfactory progenitors are not produced and the Notch signalling pathway is not activated, establishing Mash1 as a determination gene for olfactory sensory neurons. In neurogenin1 null mutant mice, olfactory progenitors are generated but they express only a subset of their normal repertoire of regulatory molecules and their differentiation is blocked. Thus neurogenin1 is required for the activation of one of several parallel genetic programs functioning downstream of Mash1 in the differentiation of olfactory sensory neurons. These results illustrate the versatility of neural bHLH genes which adopt either a determination or a differentiation function, depending primarily on the timing of their expression in neural progenitors. PMID- 11934854 TI - Insufficient VEGFA activity in yolk sac endoderm compromises haematopoietic and endothelial differentiation. AB - Vascular endothelial growth factor A (VEGFA) plays a pivotal role in the first steps of endothelial and haematopoietic development in the yolk sac, as well as in the establishment of the cardiovascular system of the embryo. At the onset of gastrulation, VEGFA is primarily expressed in the yolk sac visceral endoderm and in the yolk sac mesothelium. We report the generation and analysis of a Vegf hypomorphic allele, Vegf(lo). Animals heterozygous for the targeted mutation are viable. Homozygous embryos, however, die at 9.0 dpc because of severe abnormalities in the yolk sac vasculature and deficiencies in the development of the dorsal aortae. We find that providing 'Vegf wild-type' visceral endoderm to the hypomorphic embryos restores normal blood and endothelial differentiation in the yolk sac, but does not rescue the phenotype in the embryo proper. In the opposite situation, however, when Vegf hypomorphic visceral endoderm is provided to a wild-type embryo, the 'Vegf wild-type' yolk sac mesoderm is not sufficient to support proper vessel formation and haematopoietic differentiation in this extra-embryonic membrane. These findings demonstrate that VEGFA expression in the visceral endoderm is absolutely required for the normal expansion and organisation of both the endothelial and haematopoietic lineages in the early sites of vessel and blood formation. However, normal VEGFA expression in the yolk sac mesoderm alone is not sufficient for supporting the proper development of the early vascular and haematopoietic system. PMID- 11934855 TI - Skeletal defects in VEGF(120/120) mice reveal multiple roles for VEGF in skeletogenesis. AB - Angiogenesis is an essential component of skeletal development and VEGF signaling plays an important if not pivotal role in this process. Previous attempts to examine the roles of VEGF in vivo have been largely unsuccessful because deletion of even one VEGF allele leads to embryonic lethality before skeletal development is initiated. The availability of mice expressing only the VEGF120 isoform (which do survive to term) has offered an opportunity to explore the function of VEGF during embryonic skeletal development. Our study of these mice provides new in vivo evidence for multiple important roles of VEGF in both endochondral and intramembranous bone formation, as well as some insights into isoform-specific functions. There are two key differences in vascularization of developing bones between wild-type and VEGF(120/120) mice. VEGF(120/120) mice have not only a delayed recruitment of blood vessels into the perichondrium but also show delayed invasion of vessels into the primary ossification center, demonstrating a significant role of VEGF at both an early and late stage of cartilage vascularization. These findings are the basis for a two-step model of VEGF controlled vascularization of the developing skeleton, a hypothesis that is supported by the new finding that VEGF is expressed robustly in the perichondrium and surrounding tissue of cartilage templates of future bones well before blood vessels appear in these regions. We also describe new in vivo evidence for a possible role of VEGF in chondrocyte maturation, and document that VEGF has a direct role in regulating osteoblastic activity based on in vivo evidence and organ culture experiments. PMID- 11934856 TI - The role of smooth muscle in regulating prostatic induction. AB - We have examined the role that smooth muscle plays during prostatic organogenesis and propose that differentiation of a smooth muscle layer regulates prostatic induction by controlling mesenchymal/epithelial interactions. During development of the rat reproductive tract, an area of condensed mesenchyme involved in prostatic organogenesis is formed. This mesenchyme (the ventral mesenchymal pad, VMP) is found in both males and females, yet only males develop a prostate. We demonstrate that a layer of smooth muscle differentiates between the VMP and the urethral epithelium, and that there is a sexually dimorphic difference in the development of this layer. Serial section reconstruction showed that the layer formed at approximately embryonic day 20.5 in females, but did not form in males. In cultures of female reproductive tracts, testosterone was able to regulate the thickness of this layer resulting in a 2.4-fold reduction in thickness. We observed that prostatic buds were present in some female reproductive tracts, and determined that testosterone was able to stimulate prostatic organogenesis, depending upon the bud position relative to the smooth muscle layer. In vitro recombination experiments demonstrated that direct contact with the VMP led to the induction of very few epithelial buds, and that androgens dramatically increased bud development. Taken together, our data suggest that differentiation of a smooth muscle layer regulates signalling between mesenchyme and epithelium, and comprises part of the mechanism regulating prostatic induction. PMID- 11934857 TI - TGFbeta2 mediates the effects of hedgehog on hypertrophic differentiation and PTHrP expression. AB - The development of endochondral bones requires the coordination of signals from several cell types within the cartilage rudiment. A signaling cascade involving Indian hedgehog (Ihh) and parathyroid hormone related peptide (PTHrP) has been described in which hypertrophic differentiation is limited by a signal secreted from chondrocytes as they become committed to hypertrophy. In this negative feedback loop, Ihh inhibits hypertrophic differentiation by regulating the expression of Pthrp, which in turn acts directly on chondrocytes in the growth plate that express the PTH/PTHrP receptor. Previously, we have shown that PTHrP also acts downstream of transforming growth factor beta (TGFbeta) in a common signaling cascade to regulate hypertrophic differentiation in embryonic mouse metatarsal organ cultures. As members of the TGFbeta superfamily have been shown to mediate the effects of Hedgehog in several developmental systems, we proposed a model where TGFbeta acts downstream of Ihh and upstream of PTHrP in a cascade of signals that regulate hypertrophic differentiation in the growth plate. This report tests the hypothesis that TGFbeta signaling is required for the effects of Hedgehog on hypertrophic differentiation and expression of PTHRP: We show that Sonic hedgehog (Shh), a functional substitute for Ihh, stimulates expression of Tgfb2 and Tgfb3 mRNA in the perichondrium of embryonic mouse metatarsal bones grown in organ cultures and that TGFbeta signaling in the perichondrium is required for inhibition of differentiation and regulation of Pthrp expression by Shh. The effects of Shh are specifically dependent on TGFbeta2, as cultures from Tgfb3-null embryos respond to Shh but cultures from Tgfb2-null embryos do not. Taken together, these data suggest that TGFbeta2 acts as a signal relay between Ihh and PTHrP in the regulation of cartilage hypertrophic differentiation. PMID- 11934858 TI - Pelle kinase is activated by autophosphorylation during Toll signaling in Drosophila. AB - The Drosophila Pelle kinase plays a key role in the evolutionarily conserved Toll signaling pathway, but the mechanism responsible for its activation has been unknown. We present in vivo and in vitro evidence establishing an important role for concentration-dependent autophosphorylation in the signaling process. We first show that Pelle phosphorylation can be detected transiently in early embryos, concomitant with activation of signaling. Importantly, Pelle phosphorylation is enhanced in a gain-of-function Toll mutant (Toll(10b)), but decreased by loss-of-function Toll alleles. Next we found that Pelle is phosphorylated in transfected Schneider L2 cells in a concentration-dependent manner such that significant modification is observed only at high Pelle concentrations, which coincide with levels required for phosphorylation and activation of the downstream target, Dorsal. Pelle phosphorylation is also enhanced in L2 cells co-expressing Toll(10b), and is dependent on Pelle kinase activity. In vitro kinase assays revealed that recombinant, autophosphorylated Pelle is far more active than unphosphorylated Pelle. Importantly, unphosphorylated Pelle becomes autophosphorylated, and activated, by incubation at high concentrations. We discuss these results in the context of Toll-like receptor mediated signaling in both flies and mammals. PMID- 11934859 TI - Regulation of avian cardiogenesis by Fgf8 signaling. AB - The avian heart develops from paired primordia located in the anterior lateral mesoderm of the early embryo. Previous studies have found that the endoderm adjacent to the cardiac primordia plays an important role in heart specification. The current study provides evidence that fibroblast growth factor (Fgf) signaling contributes to the heart-inducing properties of the endoderm. Fgf8 is expressed in the endoderm adjacent to the precardiac mesoderm. Removal of endoderm results in a rapid downregulation of a subset of cardiac markers, including Nkx2.5 and Mef2c. Expression of these markers can be rescued by supplying exogenous Fgf8. In addition, application of ectopic Fgf8 results in ectopic expression of cardiac markers. Expression of cardiac markers is expanded only in regions where bone morphogenetic protein (Bmp) signaling is also present, suggesting that cardiogenesis occurs in regions exposed to both Fgf and Bmp signaling. Finally, evidence is presented that Fgf8 expression is regulated by particular levels of Bmp signaling. Application of low concentrations of Bmp2 results in ectopic expression of Fgf8, while application of higher concentrations of Bmp2 result in repression of Fgf8 expression. Together, these data indicate that Fgf signaling cooperates with Bmp signaling to regulate early cardiogenesis. PMID- 11934860 TI - LvDelta is a mesoderm-inducing signal in the sea urchin embryo and can endow blastomeres with organizer-like properties. AB - Signals from micromere descendants play a critical role in patterning the early sea urchin embryo. Previous work demonstrated a link between the induction of mesoderm by micromere descendants and the Notch signaling pathway. In this study, we demonstrate that these micromere descendants express LvDelta, a ligand for the Notch receptor. LvDelta is expressed by micromere descendants during the blastula stage, a time when signaling has been shown to occur. By a combination of embryo microsurgery, mRNA injection and antisense morpholino experiments, we show that expression of LvDelta by micromere descendants is both necessary and sufficient for the development of two mesodermal cell types, pigment cells and blastocoelar cells. We also demonstrate that LvDelta is expressed by macromere descendants during mesenchyme blastula and early gastrula stages. Macromere-derived LvDelta is necessary for blastocoelar cell and muscle cell development. Finally, we find that expression of LvDelta is sufficient to endow blastomeres with the ability to function as a vegetal organizing center and to coordinate the development of a complete pluteus larva. PMID- 11934861 TI - ASYMMETRIC LEAVES1 reveals knox gene redundancy in Arabidopsis. AB - The shoot apical meristem comprises undifferentiated stem cells and their derivatives, which include founder cells for lateral organs such as leaves. Meristem maintenance and lateral organ specification are regulated in part by negative interactions between the myb domain transcription factor ASYMMETRIC LEAVES1, which is expressed in lateral organ primordia, and homeobox transcription factors which are expressed in the shoot apical meristem (knox genes). The knox gene SHOOT MERISTEMLESS (STM) negatively regulates ASYMMETRIC LEAVES1 (AS1) which, in turn, negatively regulates other knox genes including KNAT1 and KNAT2, and positively regulates the novel gene LATERAL ORGAN BOUNDARIES (LOB). Genetic interactions with a second gene, ASYMMETRIC LEAVES2 (AS2), indicate it acts at the same position in this hierarchy as AS1. We have used a second-site suppressor screen to isolate mutations in KNAT1 and we show that KNAT1 is partially redundant with STM in regulating stem cell function. Mutations in KNAT2 show no such interaction. We discuss the regulation and evolution of redundancy among knox genes. PMID- 11934862 TI - Distal-less and homothorax regulate multiple targets to pattern the Drosophila antenna. AB - The Drosophila antenna is a highly derived appendage required for a variety of sensory functions including olfaction and audition. To investigate how this complex structure is patterned, we examine the specific functions of genes required for antenna development. The nuclear factors, Homothorax, Distal-less and Spineless, are each required for particular aspects of antennal fate. Coexpression of Homothorax, necessary for nuclear localization of its ubiquitously expressed partner Extradenticle, with Distal-less is required to establish antenna fate. Here we test which antenna patterning genes are targets of Homothorax, Distal-less and/or Spineless. We report that the antennal expression of dachshund, atonal, spalt, and cut requires Homothorax and/or Distal less, but not Spineless. We conclude that Distal-less and Homothorax specify antenna fates via regulation of multiple genes. We also report for the first time phenotypic consequences of losing either dachshund or spalt and spalt-related from the antenna. We find that dachshund and spalt/spalt-related are essential for proper joint formation between particular antennal segments. Furthermore, the spalt/spalt-related null antennae are defective in hearing. Hearing defects are also associated with the human diseases Split Hand/Split Foot Malformation and Townes-Brocks Syndrome, which are linked to human homologs of Distal-less and spalt, respectively. We therefore propose that there are significant genetic similarities between the auditory organs of humans and flies. PMID- 11934863 TI - Spitz/EGFr signalling via the Ras/MAPK pathway mediates the induction of bract cells in Drosophila legs. AB - In the development of Drosophila, the activation of the EGFr pathway elicits different cellular responses at different times and in different tissues. A variety of approaches have been used to identify the mechanisms that confer this response specificity. We have analysed the specification of bract cells in Drosophila legs. We observed that mechanosensory bristles induced bract fate in neighbouring epidermal cells, and that the RAS/MAPK pathway mediated this induction. We have identified Spitz and EGFr as the ligand and the receptor of this signalling, and by ubiquitous expression of constitutively activated forms of components of the pathway we have found that the acquisition of bract fate is temporally and spatially restricted. We have also studied the role of the poxn gene in the inhibition of bract induction in chemosensory bristles. PMID- 11934864 TI - Dnmt3L cooperates with the Dnmt3 family of de novo DNA methyltransferases to establish maternal imprints in mice. AB - Genomic imprinting is regulated by differential methylation of the paternal and maternal genome. However, it remains unknown how parental imprinting is established during gametogenesis. In this study, we demonstrate that Dnmt3L, a protein sharing homology with DNA methyltransferases, Dnmt3a and Dnmt3b, but lacking enzymatic activity, is essential for the establishment of maternal methylation imprints and appropriate expression of maternally imprinted genes. We also show that Dnmt3L interacts with Dnmt3a and Dnmt3b and co-localizes with these enzymes in the nuclei of transfected cells, suggesting that Dnmt3L may regulate genomic imprinting via the Dnmt3 family enzymes. Consistent with this model, we show that [Dnmt3a(-/-), Dnmt3b(+/-)] mice also fail to establish maternal methylation imprints. In addition, both Dnmt3a and Dnmt3L are required for spermatogenesis. Together, our findings suggest that Dnmt3L may cooperate with Dnmt3 family methyltransferases to carry out de novo methylation of maternally imprinted genes in oocytes. PMID- 11934865 TI - Residues SFQ (173-175) in the large extracellular loop of CD9 are required for gamete fusion. AB - Gamete fusion is the fundamental first step initiating development of a new organism. Female mice with a gene knockout for the tetraspanin CD9 (CD9 KO mice) produce mature eggs that cannot fuse with sperm. However, nothing is known about how egg surface CD9 functions in the membrane fusion process. We found that constructs including CD9's large extracellular loop significantly inhibited gamete fusion when incubated with eggs but not when incubated with sperm, suggesting that CD9 acts by interaction with other proteins in the egg membrane. We also found that injecting developing CD9 KO oocytes with CD9 mRNA restored fusion competence to the resulting CD9 KO eggs. Injecting mRNA for either mouse CD9 or human CD9, whose large extracellular loops differ in 18 residues, rescued fusion ability of the injected CD9 KO eggs. However, when the injected mouse CD9 mRNA contained a point mutation (F174 to A) the gamete fusion level was reduced fourfold, and a change of three residues (173-175, SFQ to AAA) abolished CD9's activity in gamete fusion. These results suggest that SFQ in the CD9 large extracellular loop may be an active site which associates with and regulates the egg fusion machinery. PMID- 11934866 TI - Expression of CD41 on hematopoietic progenitors derived from embryonic hematopoietic cells. AB - In this study, we have characterized the early steps of hematopoiesis during embryonic stem cell differentiation. The immunophenotype of hematopoietic progenitor cells derived from murine embryonic stem cells was determined using a panel of monoclonal antibodies specific for hematopoietic differentiation antigens. Surprisingly, the CD41 antigen (alphaIIb integrin, platelet GPIIb), essentially considered to be restricted to megakaryocytes, was found on a large proportion of cells within embryoid bodies although very few megakaryocytes were detected. In clonogenic assays, more than 80% of all progenitors (megakaryocytic, granulo-macrophagic, erythroid and pluripotent) derived from embryoid bodies expressed the CD41 antigen. CD41 was the most reliable marker of early steps of hematopoiesis. However, CD41 remained a differentiation marker because some CD41( ) cells from embryoid bodies converted to CD41(+) hematopoietic progenitors, whereas the inverse switch was not observed. Immunoprecipitation and western blot analysis confirmed that CD41 was present in cells from embryoid bodies associated with CD61 (beta3 integrin, platelet GPIIIa) in a complex. Analysis of CD41 expression during ontogeny revealed that most yolk sac and aorta-gonad mesonephros hematopoietic progenitor cells were also CD41(+), whereas only a minority of bone marrow and fetal liver hematopoietic progenitors expressed this antigen. Differences in CD34 expression were also observed: hematopoietic progenitor cells from embryoid bodies, yolk sac and aorta-gonad-mesonephros displayed variable levels of CD34, whereas more than 90% of fetal liver and bone marrow progenitor cells were CD34(+). Thus, these results demonstrate that expression of CD41 is associated with early stages of hematopoiesis and is highly regulated during hematopoietic development. Further studies concerning the adhesive properties of hematopoietic cells are required to assess the biological significance of these developmental changes. PMID- 11934867 TI - Runx1 is required for zebrafish blood and vessel development and expression of a human RUNX1-CBF2T1 transgene advances a model for studies of leukemogenesis. AB - RUNX1/AML1/CBFA2 is essential for definitive hematopoiesis, and chromosomal translocations affecting RUNX1 are frequently involved in human leukemias. Consequently, the normal function of RUNX1 and its involvement in leukemogenesis remain subject to intensive research. To further elucidate the role of RUNX1 in hematopoiesis, we cloned the zebrafish ortholog (runx1) and analyzed its function using this model system. Zebrafish runx1 is expressed in hematopoietic and neuronal cells during early embryogenesis. runx1 expression in the lateral plate mesoderm co-localizes with the hematopoietic transcription factor scl, and expression of runx1 is markedly reduced in the zebrafish mutants spadetail and cloche. Transient expression of runx1 in cloche embryos resulted in partial rescue of the hematopoietic defect. Depletion of Runx1 with antisense morpholino oligonucleotides abrogated the development of both blood and vessels, as demonstrated by loss of circulation, incomplete development of vasculature and the accumulation of immature hematopoietic precursors. The block in definitive hematopoiesis is similar to that observed in Runx1 knockout mice, implying that zebrafish Runx1 has a function equivalent to that in mammals. Our data suggest that zebrafish Runx1 functions in both blood and vessel development at the hemangioblast level, and contributes to both primitive and definitive hematopoiesis. Depletion of Runx1 also caused aberrant axonogenesis and abnormal distribution of Rohon-Beard cells, providing the first functional evidence of a role for vertebrate Runx1 in neuropoiesis. To provide a base for examining the role of Runx1 in leukemogenesis, we investigated the effects of transient expression of a human RUNX1-CBF2T1 transgene [product of the t(8;21) translocation in acute myeloid leukemia] in zebrafish embryos. Expression of RUNX1-CBF2T1 caused disruption of normal hematopoiesis, aberrant circulation, internal hemorrhages and cellular dysplasia. These defects reproduce those observed in Runx1-depleted zebrafish embryos and RUNX1-CBF2T1 knock-in mice. The phenotype obtained with transient expression of RUNX1-CBF2T1 validates the zebrafish as a model system to study t(8;21)-mediated leukemogenesis. PMID- 11934868 TI - An essential role for connexin43 gap junctions in mouse coronary artery development. AB - Connexin43 knockout mice die neonatally from conotruncal heart malformation and outflow obstruction. Previous studies have indicated the involvement of neural crest perturbations in these cardiac anomalies. We provide evidence for the involvement of another extracardiac cell population, the proepicardial cells. These cells give rise to the vascular smooth muscle cells of the coronary arteries and cardiac fibroblasts in the heart. We have observed the abnormal presence of fibroblast and vascular smooth muscle cells in the infundibular pouches of the connexin43 knockout mouse heart. In addition, the connexin43 knockout mice exhibit a variety of coronary artery patterning defects previously described for neural crest-ablated chick embryos, such as anomalous origin of the coronary arteries, absent left or right coronary artery, and accessory coronary arteries. However, we show that proepicardial cells also express connexin43 gap junctions abundantly. The proepicardial cells are functionally well coupled, and this coupling is significantly reduced with the loss of connexin43 function. Further analysis revealed an elevation in the speed of cell locomotion and cell proliferation rate in the connexin43-deficient proepicardial cells. A parallel analysis of proepicardial cells in transgenic mice with dominant negative inhibition of connexin43 targeted only to neural crest cells showed none of these coupling, proliferation or migration changes. These mice exhibit outflow obstruction, but no infundibular pouches. Together these findings indicate an important role for connexin43 in coronary artery patterning, a role that probably involves the proepicardial and cardiac neural crest cells. We discuss the potential involvement of connexin43 in human cardiovascular anomalies involving the coronary arteries. PMID- 11934869 TI - Partial rescue of neural apoptosis in the Lurcher mutant mouse through elimination of tissue plasminogen activator. AB - Lurcher is a spontaneous mouse mutant characterized by premature and aberrant apoptosis in the cerebellum. The phenotype has been shown to be caused by a point mutation in the delta2 glutamate receptor subunit gene that results in a large constitutive inward current, which has proved that endogenous excitotoxicity can lead to apoptotic cell death. Additional studies have suggested a direct link between this endogenous excitotoxicity and the activation of intracellular cell death enzymes. We have previously shown that excitotoxic neuronal degeneration elicited through exogenous insults (e.g. excitotoxins, stroke) is promoted by an extracellular cascade involving the serine protease tissue plasminogen activator (tPA). However, whether it is through necrotic or apoptotic mechanisms that this excitotoxic cell death occurs has remained contested. We describe the attenuation of the Lurcher cell death progression in tPA-deficient mice. Elimination of tPA delayed the apoptotic death of Purkinje and granule neurons in Lurcher mice, and reduced the phosphorylation of Jun and the activation of caspase 8. These results indicate that not only does tPA-promoted excitotoxic cell death proceed through a receptor-mediated apoptotic pathway, but that neuronal cell death in the Lurcher mouse is facilitated by extracellular cascades in addition to the already described intracellular pathways. Finally, these findings suggest that therapeutic benefits may be achieved for a wide variety of insults to the CNS by regulating tPA activity to preserve neuronal viability. PMID- 11934870 TI - Localization of intestinal intraepithelial T lymphocytes involves regulation of alphaEbeta7 expression by transforming growth factor-beta. AB - Induction of alphaEbeta7 expression on T cells by transforming growth factor (TGF)-beta is thought to be important for intestinal intraepithelial T lymphocyte (IEL) entry into the epithelial compartment. However, there has been no in vivo evidence that up-regulation of alphaEbeta7 expression on T cells by TGF-beta is critical for the selective localization of intestinal IEL in the epithelial area. We have recently established transgenic mice expressing Smad7 under the control of a distal lck promoter where TGF-beta/Smad signaling is specifically blocked in mature T cells. Here we showed that TGF-beta-mediated up-regulation of alphaEbeta7 was impaired on T cells isolated from the Smad7 transgenic mice associated with reduced numbers of intestinal IEL when compared with that in wild type littermates. These results indicated that failure to induce alphaEbeta7 on T cells by TGF-beta resulted in reduced numbers of intestinal IEL, suggesting the importance of alphaEbeta7 expression by TGF-beta in selective localization of intestinal IEL. PMID- 11934871 TI - Mutant MHC class I molecules define interactions between components of the peptide-loading complex. AB - Class I histocompatibility molecules, consisting of a heavy chain, beta2 microglobulin and peptide, are assembled in the endoplasmic reticulum (ER) with the assistance of several molecular chaperones and accessory proteins. Peptide binding occurs when assembling class I molecules associate with a loading complex consisting of the transporter associated with antigen processing (TAP) peptide transporter, tapasin, ERp57 and calreticulin (CRT)/calnexin. To assess the physical organization of this complex, we generated a series of mutants in the murine H-2Dd heavy chain and assessed their association with components of the complex. Seven mutations, clustered between amino acids 122 and 136 in the heavy chain alpha2 domain plus one mutation at position 222 in the alpha3 domain, resulted in loss of interaction with tapasin. Association with TAP was always lost simultaneously, supporting the view that tapasin acts as an obligatory bridge between class I molecules and TAP. Compared with previous studies on the HLA-A2 molecule, some differences in points of tapasin interaction were observed. Failure of the H-2Dd mutants to bind tapasin resulted in low cell-surface expression and altered intracellular transport. Most mutants retained a substantial degree of peptide loading, consistent with the view that although tapasin may promote peptide binding to class I, it is not required. A surprising observation was that all mutants lacking tapasin interaction retained normal association with CRT. This contrasts with previous observations on other class I molecules and, combined with differences in tapasin interaction, suggests that the organization of the ER peptide-loading complex can vary depending on the specific class I molecule examined. PMID- 11934872 TI - Analysis of a conformational B cell epitope of human thyroid peroxidase: identification of a tyrosine residue at a strategic location for immunodominance. AB - Thyroid peroxidase (TPO) is involved in autoimmune thyroid diseases and high titers of TPO autoantibodies directed to various conformational B cell epitopes are frequently present in patients' sera. Deciphering these epitopes is a difficult task, but can give insight into the structural basis of autoimmune recognition. TPO is a membrane-bound enzyme with the extracellular part organized in three protein domains, but of unknown three-dimensional structure. We previously localized a TPO B cell epitope within amino acid residues 742-848, a region encompassing the two C-terminal, extracellular domains of the protein. We found that at least one of the three tyrosine residues of the peptide 742-848 might be involved in autoantibody binding. In this study, we show by site directed mutagenesis that the autoepitope contains tyrosine 772 located near the hinge area between the two protein domains, suggesting they are both involved in the epitope structure. The B cell epitopes of TPO are clustered in two overlapping immunodominant regions. To map the newly localized epitope with respect of these regions, competition experiments were performed using a reference panel of TPO mAb and a further mAb previously found to be specific for the TPO peptide 742-848 at variance with all the other ones. Here, we show that the tyrosine 772-bearing epitope in the peptide 742-848 maps in a region that partly overlaps the reported two immunodominant regions. These results are suggestive of a complex TPO folding that involves all the three TPO protein domains to form a highly conformational immunodominant region. PMID- 11934873 TI - IFN-alpha enhances CD40 ligand-mediated activation of immature monocyte-derived dendritic cells. AB - Type I IFN are immune modulatory cytokines that are secreted during early stages of infection. Type I IFN bridge the innate and the adaptive immune system in humans and mice. We compared the capacity of type I and II IFN to induce the functional maturation of monocyte-derived dendritic cells (MoDC). Extending our earlier observation that type I IFN promote DC maturation, we report that these cytokines also enhance DC differentiation by augmenting CD40 ligand (CD40L) induced cytokine secretion by MoDC. Type I IFN alone were poor inducers of MoDC maturation as compared with other stimuli. They up-regulated the expression of HLA-DR, CD80, CD86, partially CCR7 but not CD83, partially reduced antigen-uptake function, increased the levels of IL-12p35 mRNA, and prolonged surface expression of peptide-MHC class I complexes for presentation to cytotoxic T lymphocytes, but did not induce migration towards CCL21 chemokine. However, type I IFN were potent co-factors for CD40L-mediated function. Here, they enhanced CD40L-mediated IL-6, IL-10 and IL-12p70 secretion. Furthermore, when combined with IL-1beta and/or IL 4, IFN-alpha2a type I IFN increased CD40L-mediated IL-12p70 production by 2- to 3 fold, and biased the IL-12 p40/p70 ratio towards the IFN-gamma inducing p70 heterodimer, this correlating with higher levels of IFN-gamma secretion by allogeneic T cell subsets and NK cells. Our results suggest that the rapid expression of CD40L, IFN and IL-1beta at sites of infection and inflammation can act in concert on immature DC, thereby linking innate and adaptive immune responses. In this way, type I IFN play a dual role as DC maturation factors and enhancers of CD40L-mediated DC activation. PMID- 11934874 TI - Bone marrow-derived mast cell differentiation is strongly reduced in histidine decarboxylase knockout, histamine-free mice. AB - Mast cells are differentiated in vitro from bone marrow precursors. In this study the development of bone marrow-derived mast cells was examined from histidine decarboxylase deficient (HDC-/-) and wild-type mice in the presence of IL-3. The number of non-adherent, tryptase- and c-kit-positive mast cells in bone marrow derived cultures of HDC(-/-) mice was decreased compared to that of wild-type (HDC+/+) animals, but within the tryptase- and c-kit-positive cells there was no difference in the expression intensity of both markers between the two groups. Furthermore, less serine proteases mMCP5, mMCP6 and FcepsilonRIalpha mRNA were detected in bone marrow-derived cell cultures originating from HDC-/- mice. Antigen-provoked degranulation through high-affinity FcepsilonI receptor was also lower in HDC-/- mice. The colony assays in semisolid medium yielded a significantly lower ratio of mixed colonies and higher proportion of macrophage colonies from HDC-/- mice-derived bone marrow compared to the wild-type. In the course of the differentiation of HDC-/- --derived mast cells exogenously added histamine is unable to substitute the endogenously missing histamine. Concordantly, alpha-fluoromethyl-histamine, the specific inhibitor of HDC, revealed only a marginal inhibition on the differentiation of tryptase-positive mast cells from wild-type mice. These findings suggest that the effect of histamine on the IL-3-dependent development of bone marrow-derived mast cell differentiation during the early period is crucial and irreplaceable. PMID- 11934875 TI - Expression and characterization of recombinant soluble human CD3 molecules: presentation of antigenic epitopes defined on the native TCR-CD3 complex. AB - The TCR-CD3 complex consists of the clonotypic disulfide-linked TCRalphabeta or TCRdeltagamma heterodimers, and the invariant CD3delta, epsilon, gamma and zeta chains. We generated plasmid constructs expressing the extracellular domains of the CD3delta, epsilon or gamma subunits fused to human IgG1 Fc. Recombinant fusion proteins consisting of individual CD3delta, epsilon or gamma subunits reacted poorly with anti-CD3 mAb including G19-4, BC3, OKT3 and 64.1. Co expression of the CD3epsilon-Ig with either the CD3delta-Ig (CD3epsilondelta-Ig) or the CD3gamma-Ig (CD3epsilongamma-Ig) resulted in fusion proteins with much increased binding to G19-4. A brief acid treatment of the purified CD3epsilondelta-Ig fusion protein substantially improved its binding to BC3, OKT3 and 64.1. Surface plasmon resonance analysis revealed that the dissociation constants for CD3epsilondelta-Ig and anti-CD3 mAb ranged from 10(-8) to 10(-9) M. Based on these results, a single-chain (sc) construct encoding the CD3delta chain linked to the CD3epsilon chain with a flexible linker followed by human IgG1 Fc was expressed. The sc CD3deltaepsilon-scIg reacted with anti-CD3 mAb without requiring acid treatment. Moreover, anti-CD3 mAb bound CD3epsilondelta-Ig at a higher affinity than CD3epsilongamma-Ig, suggesting potential structural differences between the CD3epsilondelta and CD3epsilongamma subunits. In summary, we report the expression of soluble recombinant CD3 proteins that demonstrate structural characteristics of the native CD3 complex expressed on the T cell surface. These CD3 fusion proteins can be used to further analyze the structure of the TCR-CD3 complex, and to identify molecules that can interfere with TCR-CD3 mediated signal transduction by disrupting the interaction between CD3 and TCR subunits. PMID- 11934877 TI - IFN-alpha/beta enhances BCR-dependent B cell responses. AB - Type I interferon (IFN-I) is constitutively produced in the bone marrow (BM), and induced at sites of inflammation and following infection by viruses or microorganisms. We have previously shown that IFN-I regulates the generation and selection of normal B cell populations in the BM. In the present work, we assess the effects of IFN-I on mature B cell function by monitoring the responses of IFN alpha/beta-treated murine splenic B cells to apoptotic, mitogenic and activating stimuli. A similar analysis is performed on BM mature B cells obtained from wild type or IFN-I receptor-deficient mice. IFN-alpha/beta is shown to induce B cells to a state of partial activation characterized by the up-regulation of CD69, CD86 and CD25 molecules in the absence of either proliferation or terminal differentiation. B cells treated with IFN-alpha/beta show an increased survival and resistance to Fas-mediated apoptosis. IFN-alpha/beta also enhances B cell responses to BCR ligation such as calcium fluxes, IgM internalization, induction of activation markers and proliferation. These results indicate that in addition to its inhibitory effect on viral replication and T cell apoptosis, IFN alpha/beta plays an essential role during an inflammatory response by lowering the threshold for B cell induction, thereby promoting fast and polyclonal antibody responses. PMID- 11934876 TI - Lipocalin allergen Bos d 2 is a weak immunogen. AB - The immunological characteristics of an important group of animal-derived allergens, lipocalins, are poorly known. To explore the immunology of the lipocalin allergen Bos d 2, several mouse strains with different H-2 haplotypes were immunized with the allergen. Only the BALB/c mouse mounted a distinct humoral response against Bos d 2. The proliferative spleen cell responses of all mouse strains remained very weak. Further experiments with BALB/c mice confirmed that Bos d 2 is a weak inducer of both humoral and cellular responses, and that the responses were weaker than with the control antigens hen egg lysozyme (HEL) and tetanus toxoid. IgG subclass analyses showed that Bos d 2 was prone to favor the T(h)2 response. Although s.c. immunization using complete Freund's adjuvant favored the T(h)1-deviated immune response by lymph node cells, Bos d 2 was able to induce the production of IL-4 while the control antigen HEL did not. Epitope mapping revealed that BALB/c mice recognized one immunodominant epitope in Bos d 2, almost identical to that recognized by humans. The epitope was shown to be immunogenic in subsequent experiments. However, further studies are needed to clarify the significance of priming and stimulation doses of the immunodominant and other epitopes in Bos d 2 for the outcome of immune response against the allergen. The murine immune response against Bos d 2 closely resembled that observed in humans. The weak immunogenicity of Bos d 2 may be associated with its allergenicity. PMID- 11934878 TI - Epithelial cell-derived human beta-defensin-2 acts as a chemotaxin for mast cells through a pertussis toxin-sensitive and phospholipase C-dependent pathway. AB - Mast cells are known to accumulate at the sites of inflammation in response to chemoattractants generated in the local milieu. Since human beta-defensin-2 (hBD 2) is generated in several epithelial tissues where mast cells are present and because we have recently reported that this human antibacterial peptide induces mast cell degranulation, we thus hypothesized that hBD-2 could be a mast cell chemotaxin. Here we report that hBD-2 directly and specifically induces mast cell migration with an optimal concentration of 3 microg/ml. Checkerboard analysis showed that the migration was more chemotactic rather than chemokinetic. Moreover, Scatchard analysis using 125I-labeled hBD-2 revealed that mast cells have at least two classes of receptors, high- and low-affinity receptors, for this peptide. Moreover, the competitive binding assay suggested that hBD-2 is unlikely to utilize CCR6, a functional receptor for hBD-2-mediated dendritic and T cell migration, on mast cells. In addition, treatment of mast cells with G protein inhibitor, pertussis toxin, and phospholipase C inhibitor, U-73122, abolished the cell chemotaxis in response to hBD-2, indicating that the G protein phospholipase C signaling pathway is involved in hBD-2-induced mast cell activation. Thus, we suggest that hBD-2, which was originally believed to be involved in innate host defense, may participate in the recruitment of mast cells to inflammation foci. PMID- 11934879 TI - A direct pyrophosphatase-coupled assay provides new insights into the activation of the secreted adenylate cyclase from Bordetella pertussis by calmodulin. AB - Continuous recording of the activity of recombinant adenylate cyclase (CyaA) of Bordetella pertussis (EC ) by conductimetric determination of enzyme-coupled pyrophosphate cleavage has enabled us to define a number of novel features of the activation of this enzyme by calmodulin and establish conditions under which valid activation data can be obtained. Activation either in the presence or absence of calcium is characterized by a concentration-dependent lag phase. The rate of formation and breakdown of the activated complex can be determined from an analysis of the lag phase kinetics and is in good agreement with thermodynamic data obtained by measuring the dependence of activation on calmodulin concentration, which show that calcium increases k(on) by about 30-fold. The rate of breakdown of the activated complex, formed either in the presence or absence of calcium, has been determined by dilution experiments and has been shown to be independent of the presence of calcium. The coupled assay is established as a rapid, convenient and safe method which should be readily applicable to the continuous assays of most other enzymes that catalyze reactions in which inorganic pyrophosphate is liberated. PMID- 11934880 TI - Distinct roles of receptor phosphorylation, G protein usage, and mitogen activated protein kinase activation on platelet activating factor-induced leukotriene C(4) generation and chemokine production. AB - Platelet activating factor (PAF) interacts with cell surface G protein-coupled receptors on leukocytes to induce degranulation, leukotriene C(4) (LTC(4)) generation, and chemokine CCL2 production. Using a basophilic leukemia RBL-2H3 cell line expressing wild-type PAF receptor (PAFR) and a phosphorylation deficient mutant (mPAFR), we have previously demonstrated that receptor phosphorylation mediates desensitization of PAF-induced degranulation. Here, we sought to determine the role of receptor phosphorylation on PAF-induced LTC(4) generation and CCL2 production. We found that PAF caused a significantly enhanced LTC(4) generation in cells expressing mPAFR when compared with PAFR cells. In contrast, PAF-induced CCL2 production was greatly reduced in mPAFR cells. Pertussis toxin and U0126, which inhibit G(i) and p44/42 mitogen-activated protein kinase (ERK) activation, respectively, caused very little inhibition of PAF-induced CCL2 production (approximately 20% inhibition). In contrast, these inhibitors almost completely blocked both PAF-induced ERK phosphorylation and LTC(4) generation in PAFR cells. However, in mPAFR cells pertussis toxin only partially inhibited PAF-induced ERK phosphorylation. A Ca(2+)/calmodulin inhibitor had no effect on PAF-induced ERK phosphorylation in PAFR cells but completely blocked the response in mPAFR cells. These data demonstrate that receptor phosphorylation, which serves to desensitize PAF-induced LTC(4) generation, is required for chemokine CCL2 production. They also indicate a previously unrecognized selectivity in G protein usage and ERK activation for PAF induced responses. Whereas PAF-induced CCL2 production is, in large part, mediated independently of G(i) activation or ERK phosphorylation, LTC(4) generation requires ERK phosphorylation, which is mediated by different G proteins depending on the phosphorylation status of the receptor. PMID- 11934881 TI - Cloning of the human claudin-2 5'-flanking region revealed a TATA-less promoter with conserved binding sites in mouse and human for caudal-related homeodomain proteins and hepatocyte nuclear factor-1alpha. AB - Claudin-2 is a structural component of tight junctions in the kidneys, liver, and intestine, but the mechanisms regulating its expression have not been defined. The 5'-flanking region of the claudin-2 gene contains binding sites for intestine specific Cdx homeodomain proteins and hepatocyte nuclear factor (HNF)-1, which are conserved in human and mouse. Both Cdx1 and Cdx2 activated the claudin-2 promoter in the human intestinal epithelial cell line Caco-2. HNF-1alpha augmented the Cdx2-induced but not Cdx1-induced transcriptional activation of the human claudin-2 promoter. In mice, HNF-1alpha was required for claudin-2 expression in the villus epithelium of the ileum and within the liver but not in the kidneys, indicating an organ-specific function of HNF-1alpha in the regulation of claudin-2 gene expression. Tight junction structural components, which determine epithelial polarization and intestinal barrier function, can be regulated by homeodomain proteins that control the differentiation of the intestinal epithelium. PMID- 11934883 TI - Identification of a short linear sequence present in the C-terminal tail of the rat follitropin receptor that modulates arrestin-3 binding in a phosphorylation independent fashion. AB - The rat follitropin receptor (rFSHR) is an unusual G protein-coupled receptor in that agonist-induced activation leads to the phosphorylation of the first and third intracellular loops instead of the C-terminal tail. To determine regions of G protein-coupled receptors that affect internalization independently of phosphorylation we examined the effects of truncations of the C-terminal tail of the rFSHR on agonist-induced internalization. Our studies show that progressive truncations of a region flanked by residues 642 and 651 enhance the internalization of human follicle-stimulating hormone (hFSH). Further characterization of a mutant truncated at residue 649 (designated rFSHR-t649) and another mutant in which the 642-651 region was deleted in the context of the full length rFSHR, designated rFSHR(Delta642-651), showed that both of them internalized hFSH at rates that were 2-3 times faster than rFSHR-wild type (wt). Like rFSHR-wt, however, the internalization of hFSH mediated by rFSHR-t649 and rFSHR(Delta642-651) can be inhibited with dominant-negative mutants of the non visual arrestins or dynamin. Alanine-scanning mutagenesis of the 642-651 region suggests that the effects on internalization are not mediated by a single residue, however. In an attempt to understand the molecular basis of the enhanced internalization of hFSH mediated by these mutants we used an assay that can be readily used to assess the association of the rFSHR with the arrestin-3 in co transfected cells. Using this assay we were able to show that, when compared with rFSHR-wt, rFSHR(Delta642-651) displays an approximately 4-fold enhancement in binding affinity for arrestin-3 and an approximately 1.7-fold reduction in maximal arrestin-3 binding capacity. We conclude that a short linear sequence present in the C-terminal tail of the rFSHR (642SATHNFHARK651) that is not phosphorylated limits internalization by lowering the affinity of the rFSHR for the endogenous non-visual arrestins. PMID- 11934882 TI - Hedgehog-stimulated phosphorylation of the kinesin-related protein Costal2 is mediated by the serine/threonine kinase fused. AB - The Hedgehog (Hh) signaling molecule is required for the development of numerous tissues in Drosophila. Within the cell, Hh signal transduction utilizes a large protein complex consisting of the Fused (Fu), Costal2 (Cos2), and Cubitis interruptus (Ci) proteins, but the functional interactions between these proteins are still largely uncharacterized. Using a baculovirus system, we demonstrate that the serine/threonine kinase Fu phosphorylates the kinesin-like protein Cos2 when coexpressed with Cos2. Coexpression of Cos2 and a kinase-inactive version of Fu eliminates the majority of Cos2 phosphorylation. We then show that the primary Fu-induced phosphorylation site of Cos2 is serine 572, whereas serine 931 is phosphorylated to a lesser extent. Mutation of serine 572 to alanine eliminates most, but not all, specific phosphopeptides of Cos2 when coexpressed with Fu. We also demonstrate that the phosphorylation pattern of Cos2 produced by baculovirus coexpression with kinase-dead Fu is almost identical to the phosphorylation pattern of Cos2 isolated from unstimulated S2 cells. Finally, the phosphorylation pattern of Cos2 produced by baculovirus coinfection with wild-type Fu is almost identical to that of Cos2 isolated from S2 cells stimulated by Hh, indicating that phosphorylation of serines 572 and 931 is a genuine Hh signaling event. This study clarifies the unique functions of Fu and Cos2 in Hh signal transduction and identifies only the second known phosphorylation site of a kinesin-like molecule. PMID- 11934884 TI - Herstatin, an autoinhibitor of the human epidermal growth factor receptor 2 tyrosine kinase, modulates epidermal growth factor signaling pathways resulting in growth arrest. AB - Herstatin is an autoinhibitor of the ErbB family consisting of subdomains I and II of the human epidermal growth factor receptor 2 (ErbB-2) extracellular domain and a novel C-terminal domain encoded by an intron. Herstatin binds to human epidermal growth factor receptor 2 and to the epidermal growth factor receptor (EGFR), blocking receptor oligomerization and tyrosine phosphorylation. In this study, we characterized several early steps in EGFR activation and investigated downstream signaling events induced by epidermal growth factor (EGF) and by transforming growth factor alpha (TGF-alpha) in NIH3T3 cell lines expressing EGFR with and without herstatin. Herstatin expression decreased EGF-induced EGFR tyrosine phosphorylation and delayed receptor down-regulation despite receptor occupancy by ligand with normal binding affinity. Akt stimulation by EGF and TGF alpha, but not by fibroblast growth factor 2, was almost completely blocked in the presence of herstatin. Surprisingly, EGF and TGF-alpha induced full activation of MAPK in duration and intensity and stimulated association of the EGFR with Shc and Grb2. Although MAPK was fully stimulated, herstatin expression prevented TGF-alpha-induced DNA synthesis and EGF-induced proliferation. The herstatin-mediated uncoupling of MAPK from Akt activation was also observed in Chinese hamster ovary cells co-transfected with EGFR and herstatin. These findings show that herstatin expression alters EGF and TGF-alpha signaling profiles, culminating in inhibition of proliferation. PMID- 11934885 TI - The anchoring protein RACK1 links protein kinase Cepsilon to integrin beta chains. Requirements for adhesion and motility. AB - Integrin affinity is modulated by intracellular signaling cascades, in a process known as "inside-out" signaling, leading to changes in cell adhesion and motility. Protein kinase C (PKC) plays a critical role in integrin-mediated events; however, the mechanism that links PKC to integrins remains unclear. Here, we report that PKCepsilon positively regulates integrin-dependent adhesion, spreading, and motility of human glioma cells. PKCepsilon activation was associated with increased focal adhesion and lamellipodia formation as well as clustering of select integrins, and it is required for phorbol 12-myristate 13 acetate-induced adhesion and motility. We provide novel evidence that the scaffolding protein RACK1 mediates the interaction between integrin beta chain and activated PKCepsilon. Both depletion of RACK1 by antisense strategy and overexpression of a truncated form of RACK1 which lacks the integrin binding region resulted in decreased PKCepsilon-induced adhesion and migration, suggesting that RACK1 links PKCepsilon to integrin beta chains. Altogether, these results provide a novel mechanistic link between PKC activation and integrin mediated adhesion and motility. PMID- 11934886 TI - A proteomic approach identifies proteins in hepatocytes that bind nascent apolipoprotein B. AB - The biogenesis of apolipoprotein B is quite complex in view of its huge size, hydrophobicity, obligate association with lipids such as cholesterol and triglycerides prior to secretion, and intracellular degradation of a substantial proportion of newly synthesized molecules. Multiple proteins likely serve roles as molecular chaperones to assist in folding, assembly with lipids, and regulation of the secretion of apolipoprotein B. In these studies, we developed a strategy to isolate proteins associated with apolipoprotein B in rat livers. The purification consisted of two stages: first, microsomes were prepared from rat liver and treated with chemical cross-linkers, and second, the solubilized proteins were co-immunoprecipitated with antibody against apolipoprotein B. We found that several proteins were cross-linked to apolipoprotein B. The proteins were digested with trypsin, and the released peptides were sequenced by tandem mass spectrometry. The sequences precisely matched 377 peptides in 99 unique proteins. We show that at least two of the identified proteins, ferritin heavy and light chains, can directly bind apolipoprotein B. These and possibly other proteins identified by this proteomic approach are novel candidates for proteins that affect apolipoprotein B during its biogenesis. PMID- 11934887 TI - The human papillomavirus 16 E6 protein binds to tumor necrosis factor (TNF) R1 and protects cells from TNF-induced apoptosis. AB - High risk strains of human papillomavirus (HPV), such as HPV 16, cause human cervical carcinoma. The E6 protein of HPV 16 mediates the rapid degradation of p53, although this is not the only function of E6 and cannot completely explain its transforming potential. Previous work in our laboratory has demonstrated that transfection of HPV 16 E6 into the tumor necrosis factor (TNF)-sensitive LM cell line protects expressing cells from TNF-induced apoptosis in a p53-independent manner, and the purpose of this study was to determine the molecular mechanism underlying this protection. Caspase 3 and caspase 8 activation were significantly reduced in E6-expressing cells, indicating that E6 acts early in the TNF apoptotic pathway. In fact, E6 binds directly to TNF R1, as shown both by co immunoprecipitation and mammalian two-hybrid approaches. E6 requires the same C terminal portion of TNF R1 for binding as does TNF R1-associated death domain, and TNF R1/TNF R1-associated death domain interactions are decreased in the presence of E6. HA-E6 also blocked cell death triggered by transfection of the death domain of TNF R1. Together, these results provide strong support for a model in which HPV E6 binding to TNF R1 interferes with formation of the death inducing signaling complex and thus with transduction of proapoptotic signals. They also demonstrate that HPV, like several other viruses, has developed a method for evading the TNF-mediated host immune response. PMID- 11934888 TI - The phosphorylated form of the ORF3 protein of hepatitis E virus interacts with its non-glycosylated form of the major capsid protein, ORF2. AB - Hepatitis E virus (HEV) is a human RNA virus containing three open reading frames. Of these, ORF1 encodes the viral nonstructural polyprotein; ORF2 encodes the major capsid protein, which exists in a glycosylated and non-glycosylated form; and ORF3 codes for a phosphoprotein of undefined function. Using fluorescence-based colocalization, yeast two-hybrid experiments, transiently transfected COS-1 cell co-immunoprecipitation, and cell-free coupled transcription-translation techniques, we have shown that the ORF3 protein interacts with the ORF2 protein. The domains involved in this ORF2-ORF3 association have been identified and mapped. Our deletion analysis showed that a 25-amino acid region (residues 57-81) of the ORF3 protein is required for this interaction. Using a Mexican HEV isolate, site-directed mutagenesis of ORF3, and a phosphatase digestion assay, we showed that the ORF2-ORF3 interaction is dependent upon the phosphorylation at Ser(80) of ORF3. Finally, using COS-1 cell immunoprecipitation experiments, we found that the phosphorylated ORF3 protein preferentially interacts with the non-glycosylated ORF2 protein. These findings were confirmed using tunicamycin inhibition, point mutants, and deletion mutants expressing only non-glycosylated ORF2. ORF3 maps in the structural region of the HEV genome and now interacts with the major capsid protein, ORF2, in a post translational modification-dependent manner. Such an interaction of ORF2 with ORF3 suggests a possible well regulated role for ORF3 in HEV structural assembly. PMID- 11934889 TI - Crystals of tryptophan indole-lyase and tyrosine phenol-lyase form stable quinonoid complexes. AB - The binding of substrates and inhibitors to wild-type Proteus vulgaris tryptophan indole-lyase and to wild type and Y71F Citrobacter freundii tyrosine phenol-lyase was investigated in the crystalline state by polarized absorption microspectrophotometry. Oxindolyl-lalanine binds to tryptophan indole-lyase crystals to accumulate predominantly a stable quinonoid intermediate absorbing at 502 nm with a dissociation constant of 35 microm, approximately 10-fold higher than that in solution. l-Trp or l-Ser react with tryptophan indole-lyase crystals to give, as in solution, a mixture of external aldimine and quinonoid intermediates and gem-diamine and external aldimine intermediates, respectively. Different from previous solution studies (Phillips, R. S., Sundararju, B., & Faleev, N. G. (2000) J. Am. Chem. Soc. 122, 1008-1114), the reaction of benzimidazole and l-Trp or l-Ser with tryptophan indole-lyase crystals does not result in the formation of an alpha-aminoacrylate intermediate, suggesting that the crystal lattice might prevent a ligand-induced conformational change associated with this catalytic step. Wild-type tyrosine phenol-lyase crystals bind l-Met and l-Phe to form mixtures of external aldimine and quinonoid intermediates as in solution. A stable quinonoid intermediate with lambda(max) at 502 nm is accumulated in the reaction of crystals of Y71F tyrosine phenol-lyase, an inactive mutant, with 3-F-l-Tyr with a dissociation constant of 1 mm, approximately 10-fold higher than that in solution. The stability exhibited by the quinonoid intermediates formed both by wild-type tryptophan indole-lyase and by wild type and Y71F tyrosine phenol-lyase crystals demonstrates that they are suitable for structural determination by x-ray crystallography, thus allowing the elucidation of a key species of pyridoxal 5'-phosphate-dependent enzyme catalysis. PMID- 11934890 TI - Dehydroepiandrosterone activates endothelial cell nitric-oxide synthase by a specific plasma membrane receptor coupled to Galpha(i2,3). AB - The adrenal steroid dehydroepiandrosterone (DHEA) has no known cellular receptor or unifying mechanism of action, despite evidence suggesting beneficial vascular effects in humans. Based on previous data from our laboratory, we hypothesized that DHEA binds to specific cell-surface receptors to activate intracellular G proteins and endothelial nitric-oxide synthase (eNOS). We now pharmacologically characterize a putative plasma membrane DHEA receptor and define its associated G proteins. The [3H]DHEA binding to isolated plasma membranes from bovine aortic endothelial cells was of high affinity (K(d) = 48.7 pm) and saturable (B(max) = 500 fmol/mg protein). Structurally related steroids failed to compete with DHEA for binding. The putative DHEA receptor was functionally coupled to G-proteins, because guanosine 5'-O-(3-thio)triphosphate (GTPgammaS) inhibited [3H]DHEA binding to plasma membranes by 69%, and DHEA increased [35S]GTPgammaS binding by 157%. DHEA stimulated [35S]GTPgammaS binding to Galpha(i2) and Galpha(i3), but not to Galpha(i1) or Galpha(o). Pretreatment of plasma membranes with antibody to Galpha(i2) or Galpha(i3), but not to Galpha(i1), inhibited the DHEA activation of eNOS. Thus, DHEA receptors are expressed on endothelial cell plasma membranes and are coupled to eNOS activity through Galpha(i2) and Galpha(i3). These novel findings should allow us to isolate the putative receptor and reevaluate the physiological role of DHEA activity. PMID- 11934891 TI - SDT1/SSM1, a multicopy suppressor of S-II null mutant, encodes a novel pyrimidine 5'-nucleotidase. AB - SDT1 (suppressor of disruption of TFIIS 1, YGL224c, also known as SSM1, suppressor of S-II null mutant 1) is Saccharomyces cerevisiae gene identified as a multicopy suppressor of 6-azauracil sensitivity in a null mutant of the transcription elongation factor S-II. We found that overproduction of SDT1 caused hyposensitivity to not only 6-azauracil but also 5-fluorouracil and 5 fluorocytosine. This hyposensitivity was limited to pyrimidine derivatives, and no effect was observed for non-pyrimidine drugs including such clinically used anti-fungal drugs as amphotericin B and fluconazole. Purified recombinant SDT1 protein specifically dephosphorylated 5'-UMP and 5'-CMP. These results suggested that SDT1 conferred pyrimidine-specific hyposensitivity by dephosphorylating active metabolites of 6- or 5-modified pyrimidines, i.e. 6- or 5-modified UMP. This is the first description of a highly specific pyrimidine 5'-nucleotidase in S. cerevisiae. PMID- 11934892 TI - Biochemical properties of the PsbS subunit of photosystem II either purified from chloroplast or recombinant. AB - The biochemical properties of PsbS protein, a nuclear-encoded Photosystem II subunit involved in the high energy quenching of chlorophyll fluorescence, have been studied using preparations purified from chloroplasts or obtained by overexpression in bacteria. Despite the homology with chlorophyll a/b/xanthophyll binding proteins of the Lhc family, native PsbS protein does not show any detectable ability to bind chlorophylls or carotenoids in conditions in which Lhc proteins maintain full pigment binding. The recombinant protein, when refolded in vitro in the presence of purified pigments, neither binds chlorophylls nor xanthophylls, differently from the homologous proteins LHCII, CP26, and CP29 that refold into stable pigment-binding complexes. Thus, it is concluded that if PsbS is a pigment-binding protein in vivo, the binding mechanism must be different from that present in other Lhc proteins. Primary sequence analysis provides evidence for homology of PsbS helices I and III with the central 2-fold symmetric core of chlorophyll a/b-binding proteins. Moreover, a structural homology owed to the presence of acidic residues in each of the two lumen-exposed loops is found with the dicyclohexylcarbodiimide/Ca(2+)-binding domain of CP29. Consistently, both native and recombinant PsbS proteins showed [(14)C]dicyclohexylcarbodiimide binding, thus supporting a functional basis for its homology with CP29 on the lumen-exposed loops. This domain is suggested to be involved in sensing low luminal pH. PMID- 11934893 TI - Iron-sulfur cluster biosynthesis. Thermatoga maritima IscU is a structured iron sulfur cluster assembly protein. AB - Genetic evidence has indicated that Isc proteins play an important role in iron sulfur cluster biogenesis. In particular, IscU is believed to serve as a scaffold for the assembly of a nascent iron-sulfur cluster that is subsequently delivered to target iron-sulfur apoproteins. We report the characterization of an IscU from Thermatoga maritima, an evolutionarily ancient hyperthermophilic bacterium. The stabilizing influence of a D40A substitution allowed characterization of the holoprotein. Mossbauer (delta = 0.29 +/- 0.03 mm/s, DeltaE(Q) = 0.58 +/- 0.03 mm/s), UV-visible absorption, and circular dichroism studies of the D40A protein show that T. maritima IscU coordinates a [2Fe-2S]2+ cluster. Thermal denaturation experiments demonstrate that T. maritima IscU is a thermally stable protein with a thermally unstable cluster. This is also the first IscU type domain that is demonstrated to possess a high degree of secondary and tertiary structure. CD spectra indicate 36.7% alpha-helix, 13.1% antiparallel beta-sheet, 11.3% parallel beta-sheet, 20.2% beta-turn, and 19.1% other at 20 degrees C, with negligible spectral change observed at 70 degrees C. Cluster coordination also has no effect on the secondary structure of the protein. The dispersion of signals in 1H-15N heteronuclear single quantum correlation NMR spectra of wild type and D40A IscU supports the presence of significant tertiary structure for the apoprotein, consistent with a scaffolding role, and is in marked contrast to other low molecular weight Fe-S proteins where cofactor coordination is found to be necessary for proper protein folding. Consistent with the observed sequence homology and proposed conservation of function for IscU-type proteins, we demonstrate T. maritima IscU-mediated reconstitution of human apoferredoxin. PMID- 11934894 TI - Unique ability of integrin alpha(v)beta 3 to support tumor cell arrest under dynamic flow conditions. AB - Shear-resistant arrest of circulating tumor cells is required for metastasis from the blood stream. Arrest during blood flow can be supported by tumor cell interaction with attached, activated platelets. This is mediated by tumor cell integrin alpha(v)beta3 and cross-linking plasma protein ligands. To analyze the mechanism of tumor cell ligand interactions under dynamic flow conditions, we used real-time video microscopy and tested human melanoma cell binding to fibrinogen, von Willebrand Factor, or fibronectin matrices in a buffer perfusion system. When perfused at venous flow, melanoma cells arrested abruptly and began to spread immediately. This was uniquely mediated by integrin alpha(v)beta3 on all tested ligands, and required alpha(v)beta3 activation and actin polymerization. Under static conditions, alpha(v)beta3 cooperated with alpha(v)beta1 and alpha5beta1 in supporting melanoma cell adhesion to fibronectin. But even when activated, beta1 integrins did not contribute to melanoma cell arrest during flow. Soluble ligand served as a cross-linker between attached and circulating tumor cells and enhanced melanoma cell arrest. Cohesion of activated melanoma cells was restricted to the matrix surface and did not occur in suspension. We conclude that the presence of alpha(v)beta3 in a functionally activated state provides a unique advantage for circulating tumor cells by promoting tumor cell arrest in the presence of flow-dependent shear forces. PMID- 11934895 TI - Regulation of retinoic acid-induced inhibition of AP-1 activity by orphan receptor chicken ovalbumin upstream promoter-transcription factor. AB - Retinoids are therapeutically effective in the treatment of various cancers, and some of the therapeutic action of retinoids can be ascribed to their potent inhibition of AP-1 activity that regulates transcription of genes associated with cell growth. We recently reported that the expression of orphan receptor chicken ovalbumin upstream promoter-transcription factor (COUP-TF) plays a role in mediating the growth inhibitory effect of trans-retinoic acid (trans-RA) in cancer cells. To gain insight into the molecular mechanism by which COUP-TF regulates trans-RA activity, we evaluated the effect of COUP-TF on antagonism of AP-1 activity by trans-RA. Our results demonstrated a positive correlation between COUP-TF expression and the ability of trans-RA to inhibit AP-1 activity in various cancer cell lines. In transient transfection assay, expression of COUP TF strongly inhibited tumor promoter 12-O-tetradecanoylphorbol-13-acetate-induced AP-1 transactivation activity and transactivation of c-Jun/c-Fos in both a trans RA-dependent and -independent manner. In vitro studies demonstrated that the addition of COUP-TF inhibited c-Jun DNA binding through a direct protein-protein interaction that is mediated by the DNA binding domain of COUP-TF and the leucine zipper of c-Jun. Stable expression of COUP-TF in COUP-TF-negative MDA-MB231 breast cancer cells restored the ability of trans-RA to inhibit 12-O tetradecanoylphorbol-13-acetate-induced c-Jun expression. The effect of COUP-TF in enhancing the trans-RA-induced antagonism of AP-1 activity required expression of retinoic acid receptors (RARs), since stable expression of COUP-TF in COUP-TF negative HT-1376 bladder cancer cells, which do not express RARalpha and RARbeta, failed to restore trans-RA-induced AP-1 repression. Thus, COUP-TF, through its physical interaction with AP-1, promotes anticancer effects of retinoids by potentiating their anti-AP-1 activity. PMID- 11934896 TI - Zinc-mediated dimerization and its effect on activity and conformation of staphylococcal enterotoxin type C. AB - Staphylococcal enterotoxins are superantigen exotoxins that mediate food poisoning and toxic shock syndrome in humans. Despite their structural and functional similarities, superantigens display subtle differences in biological properties and modes of receptor binding as a result of zinc atoms bound differently in their crystal structures. For example, the crystal structures of the staphylococcal enterotoxins in the type C serogroup (SECs) contain a zinc atom coordinated by one aspartate and two histidine residues from one molecule and another aspartate residue from the next molecule, thus forming a dimer. This type of zinc ligation and zinc-mediated dimerization occurs in several SECs, but not in most other staphylococcal enterotoxin serogroups. This prompted us to investigate the potential importance of zinc in SEC-mediated pathogenesis. Site directed mutagenesis was used to replace SEC zinc binding ligands with alanine. SEC mutants unable to bind zinc did not have major conformational alterations although they failed to form dimers. Zinc binding was not essential for T cell stimulation, emesis, or lethality although in general the mutants were less pyrogenic. Thus the zinc atom in SECs might represent a non-functional heavy atom in an exotoxin group that has diverged from related bacterial toxins containing crucial zinc atoms. PMID- 11934897 TI - Structure and backbone dynamics of Apo- and holo-cellular retinol-binding protein in solution. AB - Retinoid-binding proteins play an important role in regulating transport, storage, and metabolism of vitamin A and its derivatives. The solution structure and backbone dynamics of rat cellular retinol-binding protein type I (CRBP) in the apo- and holo-form have been determined and compared using multidimensional high resolution NMR spectroscopy. The global fold of the protein is consistent with the common motif described for members of the intracellular lipid-binding protein family. The most relevant difference between the NMR structure ensembles of apo- and holoCRBP is the higher backbone disorder, in the ligand-free form, of some segments that frame the putative entrance to the ligand-binding site. These comprise alpha-helix II, the subsequent linker to beta-strand B, the hairpin turn between beta-strands C and D, and the betaE-betaF turn. The internal backbone dynamics, obtained from 15N relaxation data (T1, T2, and heteronuclear nuclear Overhauser effect) at two different fields, indicate several regions with significantly higher backbone mobility in the apoprotein, including the betaC betaD and betaE-betaF turns. Although apoCRBP contains a binding cavity more shielded than that of any other retinoid carrier, conformational flexibility in the portal region may assist retinol uptake. The stiffening of the backbone in the holoprotein guarantees the stability of the complex during retinol transport and suggests that targeted retinol release requires a transiently open state that is likely to be promoted by the acceptor or the local environment. PMID- 11934898 TI - Characterization of the CTD phosphatase Fcp1 from fission yeast. Preferential dephosphorylation of serine 2 versus serine 5. AB - The C-terminal domain (CTD) of RNA polymerase II undergoes extensive phosphorylation and dephosphorylation at positions Ser2 and Ser5 during the transcription cycle. A single CTD phosphatase, Fcp1, has been identified in yeast and metazoans. Here we conducted a biochemical characterization of Fcp1 from the fission yeast Schizosaccharomyces pombe. The 723-amino acid Fcp1 protein was expressed at high levels in bacteria. Recombinant Fcp1 catalyzed the metal dependent hydrolysis of para-nitrophenyl phosphate with a pH optimum of 5.5 (kcat = 2 s(-1); K(m) = 19 mm). Deletion analysis showed that 139- and 143-amino acid segments could be deleted from the N and C termini of Fcp1, respectively, without affecting phosphatase activity. A segment containing amino acids 487-580, deletion of which abolished activity, embraces a BRCT domain present in all known Fcp1 orthologs. Mutations of residues Asp170 and Asp172 abrogated Fcp1 phosphatase activity; the essential aspartates are located within a 170DXDXT172 motif that defines a superfamily of metal-dependent phosphotransferases. We exploited defined synthetic CTD phosphopeptide substrates to show for the first time that: (i) Fcp1 CTD phosphatase activity is not confined to native polymerase II and (ii) Fcp1 displays an inherent preference for a particular CTD phosphorylation array. Using equivalent concentrations (25 microm) of CTD peptides of identical amino acid sequence and phosphoserine content, which differed only in the positions of phosphoserine within the heptad, we found that Fcp1 was 10-fold more active in dephosphorylating Ser2-PO4 than Ser5-PO4. PMID- 11934899 TI - A Functional interaction between the human papillomavirus 16 transcription/replication factor E2 and the DNA damage response protein TopBP1. AB - The human papillomavirus (HPV) transcription/replication factor E2 is essential for the life cycle of HPVs. E2 protein binds to DNA target sequences in the viral long control regions to regulate transcription of the viral genome. It also enhances viral DNA replication by interacting with the viral replication factor E1 and recruiting it to the origin of replication and may also play a more direct role in replication. The cellular proteins with which E2 interacts to carry out these functions are largely unknown. To identify these proteins a yeast two hybrid screen was carried out with the transcription/replication domain of HPV16 E2. This screen identified several candidate interacting partners for E2 including TopBP1 (topoisomerase II beta-binding protein 1). TopBP1 has eight BRCA1 carboxyl-terminal domains that are found in proteins regulating the DNA damage response, transcription, and replication. Here we demonstrate that HPV16 E2 and TopBP1 interact in vitro and in vivo and that TopBP1 can enhance the ability of E2 to activate transcription and replication. This is the first time that TopBP1 has been shown to function as a transcriptional coactivator and that E2 interacts with TopBP1. Removal of the amino-terminal domain of TopBP1 abolishes coactivation of transcription and replication. This interaction may have functional consequences upon the viral life cycle. PMID- 11934900 TI - H-Ras/mitogen-activated protein kinase pathway inhibits integrin-mediated adhesion and induces apoptosis in osteoblasts. AB - We have studied the relevance of H-Ras and its downstream effectors to osteoblast functions. 1) Purified human osteoblasts highly expressed integrins beta1, alpha4, alpha5, alpha6 and the activation epitope of beta1. However, these molecules were markedly down-regulated on osteoblasts transfected with expression vector encoding fully activated H-Ras(V12), H-Ras(V12)T35S, activating Raf 1/mitogen-activated protein kinase (MAPK), or an active Raf-1 but not on cells having H-Ras(V12)Y40C, a phosphoinositide 3-kinase (PI3K)-binding mutant. 2) Although osteoblasts spontaneously adhered to fibronectin and laminin in beta1 dependent manner, the expression of H-Ras(V12) or H-Ras(V12)T35S, but not H Ras(V12)Y40C, in osteoblasts reduced their adhesion. 3) Osteoblasts bearing H Ras(V12), H-Ras(V12)T35S, or Raf-1 failed to proliferate, whereas those with H Ras(V12)Y40C proliferated well. (4) The up-regulation of Fas and down-regulation of Bcl-2 were observed in osteoblasts expressing H-Ras(V12), H-Ras(V12)T35S, or Raf-1. (5) Most of the cells having H-Ras(V12), H-Ras(V12)T35S, or Raf-1 became annexin-V(high)/propidium iodide (PI)(high or low) and terminal deoxynucleotidyl transferase-mediated dUTP nick-end labeling (TUNEL)(high)/PI(low) after 24 and 72 h incubation, respectively. Thus, we propose that H-Ras signals followed by Raf 1/MAPK pathway but not PI3K not only reduces beta(1)-mediated adhesion of osteoblasts to matrix proteins but induces apoptosis presumably via the Fas up regulation and Bcl-2 down-regulation. PMID- 11934901 TI - Structure/activity elements of the multifunctional protein, GMEB-1. Characterization of domains relevant for the modulation of glucocorticoid receptor transactivation properties. AB - GMEB-1 was initially described as a component of a 550-kDa heteromeric DNA binding complex that is involved in the modulation of two properties of glucocorticoid receptor (GR) transactivation, the dose-response curve of agonists and the partial agonist activity of antagonists. Subsequently, GMEB-1 was also found to bind to hsp27, to associate with the coactivator TIF2 in yeast cells, and to participate in Parvovirus replication. To understand these multiple activities of GMEB-1 at a molecular level, we have now determined which regions are associated with the various activities associated with the modulation of GR transactivation properties. These activities include, homooligomerization, heterooligomerization, DNA binding, binding to GR and the transcriptional cofactor CBP, and GR modulation. Complex activities such as DNA binding and GR modulation, are found to require the physical combination of those domains that would be predicted from the involved biochemical processes. We have previously documented that GMEB-1 possesses both GR modulatory and intrinsic transactivation activity. However, the domains for these two activities of GMEB-1 are found not to overlap. This separation of activities provides a structural basis for our prior biological observations that the modulation of the dose-response curve and partial agonist activity of GR complexes is independent of the total levels of gene activation by the same GR complexes. PMID- 11934903 TI - Rappaccini's progeny. PMID- 11934902 TI - c-Src-dependent transcriptional activation of TFII-I. AB - TFII-I is a multifunctional transcription factor that is also involved in signal transduction. Here we show that TFII-I undergoes a c-Src-dependent tyrosine phosphorylation on tyrosine residues 248 and 611 and translocates to the nucleus in response to growth factor signaling. Tyrosine-phosphorylated nuclear TFII-I activates a stably integrated c-fos reporter gene. Withdrawal of signal leads to diminution of nuclear TFII-I, suggesting that the signal-dependent translocation is reversible. Antibodies against either TFII-I or c-Src abrogate growth factor stimulated activation of c-fos. Consistent with the notion that tyrosine phosphorylation of TFII-I is required for its transcriptional activity, phosphorylation-deficient mutants of TFII-I fail to activate the c-fos promoter. These data demonstrate that TFII-I, through a Src-dependent mechanism, reversibly translocates from the cytoplasm to the nucleus, leading to the transcriptional activation of growth-regulated genes. PMID- 11934904 TI - Carotid endarterectomy-the evidence. PMID- 11934905 TI - Hearing loss and connexin 26. PMID- 11934907 TI - A dubious future for public health? PMID- 11934906 TI - Prions and dentistry. PMID- 11934908 TI - Can homeopaths detect homeopathic medicines by dowsing? A randomized, double blind, placebo-controlled trial. AB - Dowsing is a method of problem-solving that uses a motor automatism, amplified through a pendulum or similar device. In a homeopathic context, it is used as an aid to prescribing and as a tool to identify miasm or toxin load. A randomized double-blind trial was conducted to determine whether six dowsing homeopaths were able to distinguish between Bryonia in a 12c potency and placebo by use of dowsing alone. The homeopathic medicine Bryonia was correctly identified in 48.1% of bottle pairs (n=156; 95% confidence interval 40.2%, 56.0%; P=0.689). These results, wholly negative, add to doubts whether dowsing in this context can yield objective information. PMID- 11934909 TI - The danger of wearing an anorak. AB - Campaigns to reduce road traffic accidents have paid little attention to the way headgear could interfere with vision. Binocular visual field measurement was undertaken in six healthy volunteers wearing four different types of anorak. All four anoraks greatly reduced the horizontal and superior field of vision. The anorak producing the worst reduction resulted in a width of vision of 99 degrees and only 15 degrees of vision above eye level, versus 167 degrees and 52 degrees respectively without an anorak. Anorak wearers should turn their heads to look sideways before crossing the road. PMID- 11934910 TI - Colonoscopy at a combined district general hospital and specialist endoscopy unit: lessons from 505 consecutive examinations. AB - Provisional reports from the Intercollegiate British Society of Gastroenterology National Colonoscopy audit show completion rates of 57-77% for the procedure and poor levels of training and supervision. We prospectively audited all aspects of colonoscopy performed at a combined district general hospital and specialist endoscopy unit. Details of referral, examination, endoscopist, complications and follow-up were recorded and patients were sent questionnaires for long-term follow-up. 505 patients (246 male) underwent colonoscopy by 27 different endoscopists. Their median age was 57 years (range 13-92) and 93% were outpatients. 64% patients were symptomatic and 36% were having surveillance or follow-up colonoscopy. The overall caecal intubation rate was 93%, with little difference between surgeons, physicians and experienced trainees (89%, 92%, 94%) and specialist endoscopists (98%). In only one case was an inexperienced trainee (<100 procedures) unsupervised. Pain scores estimated by the endoscopist were well matched with those given by the patient-medians 29 and 26 (maximum 100) respectively. Median satisfaction score was 96 (maximum 100). Polyp pick-up rate was 26.9% and there were 11 new cancers. 16 (3%) minor immediate complications were recorded-5 oversedation, 6 vasovagal attacks, 3 polypectomy haemorrhages and 2 mucosal injuries (neither requiring treatment). 3 patients died within 6 months of follow-up but no death was colonoscopy related. Completion rates in this setting were adequate for all endoscopists studied. Patient satisfaction with the procedure was high and very few immediate or long-term complications were encountered. PMID- 11934911 TI - Intrathoracic migration of a Kirschner wire. PMID- 11934912 TI - Renal cell carcinoma in pregnancy. PMID- 11934913 TI - Meige's syndrome in dementia with Lewy bodies. PMID- 11934914 TI - Colonic endometriosis or adenoma? PMID- 11934915 TI - Subclinical type 1 diabetes. PMID- 11934916 TI - A young man with parkinsonism. PMID- 11934917 TI - Psychiatry, postmodernism and postnormal science. PMID- 11934918 TI - What's the point of rigorous research on complementary/alternative medicine? PMID- 11934919 TI - Oesophageal cancer surgery. PMID- 11934920 TI - Flugelmensch. PMID- 11934921 TI - Confidentiality. PMID- 11934922 TI - Teaching clinical skills to preregistration house officers. PMID- 11934923 TI - Selenium deficiency and cardiomyopathy. PMID- 11934925 TI - Self-regulation in hospital waiting lists. PMID- 11934926 TI - Self-regulation in hospital waiting lists. PMID- 11934927 TI - The Perruche judgment and the "right not to be born". AB - The French government has given in to public pressure and overturned a controversial legal ruling which recognised the right of a disabled child to seek damages. Most notably, the ruling, widely described as establishing a child's right "not to be born", had provoked "outrage" amongst groups defending the rights of the disabled and led to a ban on prenatal scans by French gynaecologists. Once again, only parents will be able to seek damages but some people think the ruling has been misinterpreted. PMID- 11934928 TI - Is there a "right not to be born"? Reproductive decision making, options and the right to information. PMID- 11934929 TI - Are doctors altruistic? AB - There is a growing belief in the US that medicine is an altruistic profession, and that physicians display altruism in their daily work. We argue that one of the most fundamental features of medical professionalism is a fiduciary responsibility to patients, which implies a duty or obligation to act in patients' best medical interests. The term that best captures this sense of obligation is "beneficence", which contrasts with "altruism" because the latter act is supererogatory and is beyond obligation. On the other hand, we offer several examples in which patients act altruistically. If it is patients and not the doctors who are altruistic, then the patients are the gift-bearers and to that extent doctors owe them gratitude and respect for their many contributions to medicine. Recognising this might help us better understand the moral significance of the doctor-patient relationship in modern medicine. PMID- 11934930 TI - Supererogation and the profession of medicine. AB - In the light of increasing public mistrust, there is an urgent need to clarify the moral status of the medical profession and of the relationship of the clinician to his/her patients. In addressing this question, I first establish the coherence, within moral philosophy generally, of the concept of supererogation (the doing of more than one's duty). I adopt the notion of an act of "unqualified" supererogation as one that is non-derivatively good, praiseworthy, and freely undertaken for others' benefit at the risk of some cost to the agent. I then argue that committing oneself to the profession of clinical medicine is an act of this kind. This is the case, not because the aim of medicine is to help patients, but because of the open ended commitment of time and the vulnerability to the consequences of failure that the clinician must accept. PMID- 11934931 TI - Commentary on Glannon and Ross, and McKay. AB - The patient-doctor relationship has recently come under intense scrutiny, resulting in a re-evaluation of the basis of that relationship. The papers by Glannon and Ross, and McKay seek to identify the sources of authority in the patient-doctor relationship by evaluating it in terms of the concept of altruism. In this paper I argue that the analysis of Glannon and Ross, and of McKay is unnecessary and that the analysis offered by the latter is also flawed. I do acknowledge, however, that Glannon and Ross's description of doctors' responsibilities and patients' roles has much to commend it. PMID- 11934932 TI - Supererogation and altruism: a comment. AB - Supererogation can be distinguished from altruism, in that the former is located in the category of duty but exceeds the strict requirements of duty, whereas altruism belongs to a different moral category from duty. It follows that doctors do not act altruistically in their professional roles. Individual doctors may sometimes show supererogation, but supererogation is not a necessary feature of the medical profession. The aim of medicine is to act in the best interests of patients. This aim involves neither supererogation nor even the moral quality of beneficence. It is simply a job description. Morality enters medicine through the quality of the individual doctor's work, not by the definition of that work. PMID- 11934933 TI - Is there a moral duty for doctors to trust patients? AB - In this paper I argue that it is morally important for doctors to trust patients. Doctors' trust of patients lays the foundation for medical relationships which support the exercise of patient autonomy, and which lead to an enriched understanding of patients' interests. Despite the moral and practical desirability of trust, distrust may occur for reasons relating to the nature of medicine, and the social and cultural context within which medical care is provided. Whilst it may not be possible to trust at will, the conscious adoption of a trusting stance is both possible and warranted as the burdens of misplaced trust fall more heavily upon patients than doctors. PMID- 11934934 TI - Reflections on a new medical cosmology. AB - Since the nineteenth century the theory and practice of mainstream Western medicine has been grounded in the biomedical model. In the later years of the twentieth century, however, it has faced a range of serious problems, which when viewed collectively, remain unresolved despite a variety of responses. The question we now face is whether these problems can be dealt with by modifying and extending the principles underlying the biomedical model, or whether a more radical solution is required. Recent critiques of Western medicine have focused mainly on the biopsychosocial model in relation to the former approach, but it will be contended that this cannot deal adequately with the challenges that medicine currently faces, because although it addresses both the scientific and humanistic aspects of medicine it fails to harmonise them. I shall therefore argue for the necessity of a more radical approach, and suggest that what is required to accomplish this is the development of a new medical cosmology, rooted in an older and more global framework. Such a fundamental change would inevitably involve a long term process which it is not yet possible to fully comprehend let alone specify in detail. Some of the necessary features of such a new medical cosmology can, however, already be distinguished and the outline of these is described. PMID- 11934935 TI - Embryonic stem cell production through therapeutic cloning has fewer ethical problems than stem cell harvest from surplus IVF embryos. AB - Restrictions on research on therapeutic cloning are questionable as they inhibit the development of a technique which holds promise for successful application of pluripotent stem cells in clinical treatment of severe diseases. It is argued in this article that the ethical concerns are less problematic using therapeutic cloning compared with using fertilised eggs as the source for stem cells. The moral status of an enucleated egg cell transplanted with a somatic cell nucleus is found to be more clearly not equivalent to that of a human being. Based on ethical considerations alone, research into therapeutic cloning should be encouraged in order to develop therapeutic applications of stem cells. PMID- 11934937 TI - Preventive detention must be resisted by the medical profession. AB - A policy of "preventive detention" has recently been debated in the British Parliament. Alarmed by the high-profile criminal activities of people suspected of having dangerous severe personality disorder (DSPD), the government have made clear their intention to "indeterminately but reviewably detain" people with DSPD, after diagnosis by forensic psychiatrists, even if the individuals are yet to commit an offence. Such a policy may improve the safety of the public, but has obvious implications for civil liberties. This essay criticises the morality of the government's intention and rejects the notion that the medical profession could ethically collude with such a policy. PMID- 11934936 TI - Death, dying and donation: organ transplantation and the diagnosis of death. AB - Refusal of organ donation is common, and becoming more frequent. In Australia refusal by families occurred in 56% of cases in 1995 in New South Wales, and had risen to 82% in 1999, becoming the most important determinant of the country's very low organ donation rate (8.9/million in 1999). Leading causes of refusal, identified in many studies, include the lack of understanding by families of brain death and its implications, and subsequent reluctance to relegate the body to purely instrumental status. It is an interesting paradox that surveys of the public continue to show considerable support for organ donation programmes--in theory we will, in practice we won't (and don't). In this paper we propose that the Australian community may, for good reason, distrust the concept of and criteria for "whole brain death", and the equation of this new concept with death of the human being. We suggest that irreversible loss of circulation should be reinstated as the major defining characteristic of death, but that brain-dead, heart-beating entities remain suitable organ donors despite being alive by this criterion. This presents a major challenge to the "dead donor rule", and would require review of current transplantation legislation. Brain dead entities are suitable donors because of irreversible loss of personhood, accurately and robustly defined by the current brain stem criteria. Even the dead are not terminally ill any more. PMID- 11934938 TI - Thomson, the right to life, and partial birth abortion or two MULES for Sister Sarah. AB - In this paper, I argue that Thomson's famous attempt to reconcile the fetus's putative right to life with robust abortion rights is not tenable. Given her view, whether or not an abortion violates the fetus's right to life depends on the abortion procedure utilised. And I argue that Thomson's view implies that any late term abortion that involves feticide is impermissible. In particular, this would rule out the partial birth abortion technique which has been so controversial of late. PMID- 11934939 TI - Two challenges to the double effect doctrine: euthanasia and abortion. AB - The validity of the double effect doctrine is examined in euthanasia and abortion. In these two situations killing is a method of treatment. It is argued that the doctrine cannot apply to the care of the dying. Firstly, doctors are obliged to harm patients in order to do good to them. Secondly, patients should make their own value judgments about being mutilated or killed. Thirdly, there is little intuitive moral difference between direct and indirect killing. Nor can the doctrine apply to abortion. Doctors kill fetuses as a means of treating the mother. They also kill them as an inevitable side effect of other treatment. Drawing a moral distinction between the direct and the indirect killing gives counterintuitive results. It is suggested that pragmatic rules, not ethics, govern practices around euthanasia and cause it to be more restricted than abortion. PMID- 11934940 TI - The ethics of surgery in the elderly demented patient with bowel obstruction. AB - OBJECTIVE: Little has been written in the medical literature concerning the ethics of treatment of the elderly demented patient with bowel obstruction. It is one example of the issues with which we are becoming increasingly involved. We conducted a survey of our colleagues' opinions to determine current practice. DESIGN: A postal questionnaire study (62% response rate). Questions were posed that related to a case scenario of an elderly demented patient presenting with a presumed sigmoid volvulus. SETTING: The northern region of England. PARTICIPANTS: Thirty seven surgical members of the Association of Coloproctology of Great Britain and Ireland, Northern Chapter. RESULTS: Sixty five per cent of respondents felt that surgery would be inappropriate, and 26% that any intervention at all upon the subject in the case scenario would be inappropriate. More would operate, however, at the request of relatives. An advance directive not to treat would be respected by 70% despite a relative's wishes. CONCLUSIONS: Overall there was a wide variation in the approach of the surgeons to a demented patient with bowel obstruction. In an era of clinical governance, and an increased awareness of the ethics of consent, this study presents one example of the difficult decisions with which we are increasingly faced. The greater use of advance directives may provide one possible solution. PMID- 11934941 TI - Factors affecting physicians' decisions to forgo life-sustaining treatments in terminal care. AB - OBJECTIVES: Treatment decisions in ethically complex situations are known to depend on a physician's personal characteristics and medical experience. We sought to study variability in decisions to withdraw or withhold specific life supporting treatments in terminal care and to evaluate the association between decisions and such background factors. DESIGN: Readiness to withdraw or withhold treatment options was studied using a terminal cancer patient scenario with alternatives. Physicians were asked about their attitudes, life values, experience, and training; sociodemographic data were also collected. SETTING: Finnish physicians, postal survey. SURVEY SAMPLE: Five hundred general practitioners, 300 surgeons, 300 internists, and 82 oncologists. RESULTS: Treatments most often forgone were blood transfusion (82%) and thrombosis prophylaxis (81%). Least willingly abandoned were intravenous (i.v.) hydration (29%) and supplementary oxygen (13%). Female doctors were less likely to discontinue thrombosis prophylaxis (p=0.022) and supplementary oxygen (p<0.001), but more readily x ray (p=0.039) and laboratory (p=0.057) examinations. Young doctors were more likely to continue antibiotics (p=0.025), thrombosis prophylaxis (p=0.006), supplementary oxygen (p=0.004) and laboratory tests (p=0.041). Oncologists comprised the specialty most ready to forgo all studied treatments except antibiotics and blood transfusion. The family's wishes (alternative 1) significantly increased treatment activity. Young and female practitioners and oncologists were most influenced by family appeal. Advance directives (alternative 2) made decisions significantly more reserved and uniform. Different factors in the physician's background were found to predict decisions to withdraw antibiotics or i.v. hydration. CONCLUSION: The considerable variation observed in doctors' decisions to forgo specific life-sustaining treatments (LST) was seen to depend on their personal background factors. Experience, supervision, and postgraduate education seemed to be associated with more reserved treatment decisions. To increase the objectivity of end of life decisions, training, and research are of prime significance in this ethically complex area of medicine. PMID- 11934943 TI - Actions, causes, and psychiatry: a reply to Szasz. AB - In a recent paper, it was argued forcefully by Thomas Szasz that it is crucial to the scientific credibility of psychiatry that it abandon talk of the behaviour of the mentally "ill" in terms of causes: such behaviour is not caused by their condition--it simply has reasons, which are discounted by the medical model. It is argued in this paper that Szasz's theory is incomplete for two reasons: first, in assuming that reasons are radically different from causes, it cannot account for the possibility that "sane" behaviour might be just as much caused as "insane"; and second, it tacitly assumes that the origin of behaviour always lies with the agent--a view that arguably is an accident of grammar. Hence while there is no mental illness, this is because there is nothing that could be ill--and this means that there is no such thing as mental "health" either. PMID- 11934942 TI - Conditions and consequences of medical futility--from a literature review to a clinical model. AB - OBJECTIVES: To present an analysis of "futility" that is useful in the clinical setting. DESIGN: Literature review. MATERIAL AND METHODS: According to Medline more than 750 articles have been published about medical futility. Three criteria (language, time period, and the authors expressed their own opinions) singled out 43 of them. The authors' opinions about futility were analysed using the scheme: "If certain conditions are satisfied, then a particular measure is futile" and "If a particular measure is futile, then certain moral consequences are implied". RESULTS: Regarding conditions, most authors stated that judgments about futility should be made by physicians. The measure was usually some kind of medical treatment, and the goals related to quality of life, physiological improvement, or prolongation of life. The probability of success in reaching the goal was in most cases described in semiquantitative terms. Regarding consequences, the authors stated that health care professionals may (sometimes ought or should) withhold or withdraw a futile measure, most often after a dialogue with the patient (29 articles), but sometimes without informing the patient (nine articles), or with one-way information (four articles). Over time more and more articles recommend that the patient should be involved in joint decision making. Based on this literature review a clinical model was developed. CONCLUSIONS: The model, requiring that conditions and consequences should be made explicit, may, in "futility situations", facilitate both the collection of the necessary information and make the moral implications visible. It also makes communication about measures considered to be futile possible without using such ambiguous terms as "futile". PMID- 11934945 TI - Attitudes of Hungarian students and nurses to physician assisted suicide. AB - In Hungary, which has one of the highest rates of suicide in the world, physician assisted suicide (PAS) and euthanasia are punishable criminal acts. Attitudes towards self destruction and assisted suicide are, however, very controversial. We investigated the attitudes of medical students, nurses and social science students in Hungary towards PAS, using a twelve item scale: the total number of participants was 242. Our results indicate a particular and controversial relationship between attitudes towards assisted suicide in Hungary and experience with terminally ill people. The social science students, who had the fewest personal experiences with terminally ill patients, are characterised by the most permissive attitudes towards assisted suicide. Nurses, who had everyday contact and experience with these patients, were the most conservative, being least supportive of assisted suicide. The attitudes of medical students, the would be physicians, are somewhere between those of nurses and social science students. PMID- 11934947 TI - Bushfires: are we doing enough to reduce the human impact? PMID- 11934948 TI - Neurobehavioural testing in workers occupationally exposed to lead: systematic review and meta-analysis of publications. AB - Although the toxic effects of lead on the central nervous system have been well described, the blood concentration at which lead begins to exert adverse effects remains the focus of debate. A meta-analysis of occupational studies was conducted evaluating the association between neurobehavioural testing results and moderate blood lead concentrations. PMID- 11934949 TI - Exposure to carbon monoxide and nitrogen dioxide in enclosed ice arenas. AB - This article summarises the latest information on the adverse cardiorespiratory effects of exposure to carbon monoxide (CO) and nitrogen dioxide (NO(2)) in enclosed ice rinks. Sources of CO and NO(2) emissions are identified, current standards for these agents, as well as methods of controlling the emissions, dispersion, and evacuation of these toxic gases are presented. A detailed literature search involving 72 references in English and French from research conducted in North America and Europe was used. Material was from peer reviewed journals and other appropriate sources. Air pollutants such as carbon monoxide (CO), and nitrogen dioxide (NO(2)) which are present in enclosed skating facilities, may exacerbate a pre-existing pathogenic condition in those people who spend considerable time in these environments. Considering the popularity of ice hockey, short track speed skating, and figure skating, and the hundreds of hours that a sensitive person may spend each year in these environments, it would seem appropriate to seek more definitive answers to this important health problem. From the findings and conclusions of the research reviewed in this paper, 10 recommendations are listed. PMID- 11934950 TI - Sperm count and chromatin structure in men exposed to inorganic lead: lowest adverse effect levels. AB - OBJECTIVES: To obtain knowledge on male reproductive toxicity of inorganic lead at current European exposure levels and to establish lowest adverse effect levels, if any. METHODS: A cross sectional survey of the semen of 503 men employed by 10 companies was conducted in the United Kingdom, Italy, and Belgium. The mean blood lead concentration was 31.0 microg/dl (range 4.6-64.5) in 362 workers exposed to lead and 4.4 microg/dl (range below the detection limit of 19.8) in 141 reference workers. Semen volume and sperm concentration were determined in a fresh semen sample according to an agreed protocol subject to quality assurance. The sperm chromatin structure assay (SCSA) was performed at a centralised laboratory. Extraneous determinants including centre, period of sexual abstinence, and age were taken into account in the statistical analysis. If appropriate, possible thresholds were examined by iterative threshold slope linear regression. RESULTS: The median sperm concentration was reduced by 49% in men with blood lead concentration above 50 microg/dl. There was no indication of a linear trend of lower sperm concentration with increasing blood lead values, but threshold slope least square regression identified a blood lead concentration of 44 microg/dl (beta=-0.037, F=4.35, p=0.038) as a likely threshold. Abnormal sperm chromatin structure was not related to blood lead concentration, but some indications of deterioration of sperm chromatin was found in men with the highest concentrations of lead within spermatozoa. Biological monitoring data did not indicate long term effects of lead on semen quantity or sperm chromatin. CONCLUSION: Adverse effects of lead on sperm concentration and susceptibility to acid induced denaturation of sperm chromatin are unlikely at blood lead concentrations below 45 microg/dl. Effects of low level exposure to lead on other measures of testicular function cannot be ruled out. PMID- 11934951 TI - Distribution and determinants of trihalomethane concentrations in indoor swimming pools. AB - OBJECTIVES: For many decades chlorination has been used as a major disinfectant process for public drinking and swimming pool water in many countries. However, there has been rising concern over the possible link between disinfectant byproducts (DBPs) and adverse reproductive outcomes. The purpose of this study was to estimate the concentrations of trihalomethanes (THMs) in some indoor swimming pools in London and their variation within and between pools and any correlation with other factors. METHODS: Water samples were collected from eight different indoor swimming pools in London. A total of 44 pool samples were collected and analysed for total organic content (TOC) and THMs. Water and air temperature were measured along with the pH during the collection of pool samples. The level of turbulence and the number of people in the pool at the time were also assessed. RESULTS: The geometric mean concentration for all swimming pools of TOC was 5.8 mg/l, of total THMs (TTHMs) 132.4 microg/l, and for chloroform 113.3 microg/l. There was a clear positive linear correlation between the number of people in the swimming pool and concentrations of TTHMs and chloroform (r=0.7, p<0.01), and a good correlation between concentrations of TOC and TTHMs (r=0.5, p<0.05) and water temperature and concentrations of TTHMs (r=0.5, p<0.01). There was a larger variation in THMs within pools than between pools. CONCLUSION: Relatively high concentrations of THMs were found in London's indoor swimming pools. The levels correlated with the number of people in the pool, water temperature, and TOC. The variation in concentrations of THMs was greater within pools than between pools. PMID- 11934952 TI - Updated epidemiological study of workers at two California petroleum refineries, 1950-95. AB - OBJECTIVES: To further assess the potential role of occupational exposures on mortality, a second update of a cohort study of workers at two petroleum refineries in California was undertaken. METHODS: Mortality analyses were based on standardised mortality ratios (SMRs) and 95% confidence intervals (95% CIs) using the general population of California as a reference. Additional analyses of lymphatic and haematopoietic cancer deaths and diseases related to asbestos were undertaken. RESULTS: The update consisted of 18,512 employees, who contributed 456,425 person-years of observation between 1950 and 1995. Both overall mortality and total cancer mortality were significantly lower than expected, as were several site specific cancers and non-malignant diseases. In particular, no significant increases were reported for leukaemia cell types or non-Hodgkin's lymphoma. Mortality excess from multiple myeloma was marginally significant. The excess was confined to employees enrolled before 1949. Furthermore, there was no significant upward trend based on duration of employment, which argues against a causal interpretation relative to employment or exposures at the refineries. No increase was found for diseases related to asbestos: pulmonary fibrosis; lung cancer; or malignant mesothelioma. There was no significant increase in mortality from any other cancers or non-malignant diseases. CONCLUSION: This second update provides additional reassurance that employment at these two refineries is not associated with increased risk of mortality. PMID- 11934953 TI - Sunlight and mortality from breast, ovarian, colon, prostate, and non-melanoma skin cancer: a composite death certificate based case-control study. AB - OBJECTIVES: To explore whether mortality from female breast, ovarian, colon, and prostate cancer were negatively associated with exposure to sunlight. METHODS: A death certificate based case-control study of mortality was conducted into five cancers: female breast, ovarian, colon, prostate, and non-melanoma skin cancer (as a positive control) to examine associations with residential and occupational exposure to sunlight. Cases were all deaths from these cancers between 1984 and 1995 in 24 states of the United States. Controls, which were age frequency matched to a series of cases, excluded deaths from cancer and certain neurological diseases. Multiple logistic regression was used in a model that included age, sex, race, residential exposure to sunlight (based on region), and socioeconomic status, occupational exposure to sunlight, and physical activity (the last three based on usual occupation). RESULTS: Residential exposure to sunlight was negatively and significantly associated with mortality from female breast, ovarian, prostate, and colon cancer. Only female breast and colon cancer, however, also showed significant negative associations with jobs with the highest occupational exposure to sunlight (odds ratio (OR) 0.82 (95% confidence interval (95% CI) 0.70 to 0.97) for female breast cancer; OR 0.90 (95% CI 0.86 to 0.94) for colon cancer). For both cancers, the negative association with occupational sunlight was greatest in the geographical region of highest exposure to sunlight and was independent of physical activity on the job. Non-melanoma skin cancer, as expected, was positively associated with both residential and occupational sunlight. CONCLUSIONS: In this exploratory study, unlike mortality from non melanoma skin cancer, mortality from female breast cancer and colon cancer were negatively associated with both residential and occupational sunlight. PMID- 11934954 TI - Psychomotor performance and subjective symptoms at low level toluene exposure. AB - OBJECTIVES: Possible effects of long term occupational exposure to toluene below the level of 100 ppm on psychomotor performance and subjective symptoms were investigated in a cross sectional approach. METHODS: From German rotogravure printing plants 278 male workers, mean age 39.8 years, mean duration of employment 14.9 years, were examined. A mean lifetime weighted average exposure (LWAE) of 45.1 ppm toluene in ambient air was found for 154 exposed workers (rotogravure printing area), with a mean current exposure of 24.7 ppm. The corresponding data for a second group of 124 workers with very low exposure (endprocessing area) had LWAE of 9.3 ppm and a current exposure of 3.3 ppm toluene. Psychomotor performance (steadiness, line tracing, aiming, tapping, and peg board) and subjective symptoms were examined. RESULTS: No significant differences between the two exposure groups were found by analysis of variance (ANOVA). By stepwise linear regression analyses there were weak associations of LWAE with one performance variable and two symptoms scales, but the results were not significant after correction for the alpha error. Psychomotor performance was mostly affected by age (maximum explained variance up to 13%), and handedness (up to 9%), whereas subjective symptoms are mostly affected by anxiety (up to 38%). CONCLUSIONS: The weak associations between long term exposure to toluene should be used to indicate further longitudinal investigations. The results of this cross sectional study show no obvious dose response relation for psychomotor functions and subjective symptoms among workers exposed to toluene at a current exposure level of 1-88 ppm. PMID- 11934956 TI - Global impact of AIDS on work. PMID- 11934955 TI - Epidemiological study to investigate potential interaction between physical and psychosocial factors at work that may increase the risk of symptoms of musculoskeletal disorder of the neck and upper limb. AB - OBJECTIVES: To investigate potential interactions between physical and psychosocial risk factors in the workplace that may be associated with symptoms of musculoskeletal disorder of the neck and upper limb. METHODS: 891 of 1514 manual handlers, delivery drivers, technicians, customer services computer operators, and general office staff reported on physical and psychosocial working conditions and symptoms of neck and upper limb disorders using a self administered questionnaire (59% return rate). Of the 869 valid questionnaire respondents, 564 workers were classified in to one of four exposure groups: high physical and high psychosocial, high physical and low psychosocial, low physical and high psychosocial, and low physical and low psychosocial. Low physical and low psychosocial was used as an internal reference group. The exposure criteria were derived from the existing epidemiological literature and models for physical and psychosocial work factors. The frequency and amplitude of lifting and the duration spent sitting while experiencing vibration were used as physical exposure criteria. Ordinal values of mental demands, job control, and social support with managers and coworkers were used as psychosocial exposure criteria. RESULTS: In the multivariate analyses, the highest and significant increase in risk was found in the high physical and high psychosocial exposure group for symptoms of hand or wrist and upper limb disorders after adjusting for years at the job, age, and sex. A potential interaction effect was found for the symptoms of the hand or wrist and upper limb disorders but not for the neck symptoms. CONCLUSION: This study showed that workers highly exposed to both physical and psychosocial workplace risk factors were more likely to report symptoms of musculoskeletal disorders than workers highly exposed to one or the other. The results suggest an interaction between physical and psychosocial risk factors in the workplace that increased the risk of reporting symptoms in the upper limbs. Psychosocial risk factors at work were more important when exposure to physical risk factors at work were high than when physical exposure was low. Ergonomic intervention strategies that aim to minimise the risks of work related musculoskeletal disorders of the upper limb should not only focus on physical work factors but also psychosocial work factors. PMID- 11934957 TI - Musculoskeletal sonography by rheumatologists: the challenges. PMID- 11934958 TI - Systemic lupus erythematosus and dysregulated apoptosis-what is the evidence? PMID- 11934959 TI - Erosions in inflammatory polyarthritis are symmetrical regardless of rheumatoid factor status: results from a primary care-based inception cohort of patients. AB - BACKGROUND AND AIMS: Symmetry is considered an important criterion for the differentiation of rheumatoid arthritis (RA) from other forms of inflammatory polyarthritis (IP), particularly those that are seronegative. Because of the inclusion of symmetry in the diagnostic and classification process, however, its true occurrence in RA cannot be assessed. As a surrogate, peripheral inflammatory arthropathies associated with rheumatoid factor production may be more likely to be symmetrical. We examined the degree of symmetry of erosions in an unselected cohort of patients with IP and tested the hypothesis that the presence of rheumatoid factor (RF) is associated with greater symmetry. METHODS: All patients registered with The Norfolk Arthritis Register (NOAR; a UK primary-care based cohort of patients with IP with annual follow-up) and who had radiographs performed at the fifth anniversary from notification were included in the analysis. Radiographs of the hands and feet were read using the Larsen method; a score of 2 or more in any particular joint indicated an erosion. Log-linear modelling was used to determine the symmetry of erosions between right and left for the following joint groups: wrists, metacarpophalangeal joints, proximal interphalangeal joints and metatarsophalangeal joints. Log-linear modelling was also used to determine the influence of RF on symmetry. RESULTS: Five hundred and thirty-seven patients contributed to the analysis. The median time to performing radiographs was 69 months (interquartile range 65.5-74.8) from the onset of symptoms. A total of 212 (39%) patients had erosive disease. Overall, IP was found to be a symmetrical disease. Despite there being more erosions in RF positive patients, there was no greater excess of symmetry in RF-positive compared with RF-negative patients. CONCLUSION: Radiographically, IP is a symmetrical disease irrespective of RF status. The use of symmetry as an important feature in identifying subgroups of patients with IP, such as RA, is challenged. PMID- 11934960 TI - Unwillingness of rheumatoid arthritis patients to risk adverse effects. AB - OBJECTIVE: To evaluate patient willingness to accept the risk of adverse effects (AEs) commonly associated with arthritis medications. METHODS: Rheumatoid arthritis patients were asked to rate their willingness to take a medication associated with 17 specific AEs using a visual analogue scale. RESULTS: We interviewed 100 patients. Eighty-one were currently using one or more disease modifying anti-rheumatic drugs (DMARDs) and 29 had previously experienced AEs related to DMARDs. Seventy-five stated that they were doing very well or well with respect to their arthritis compared with other people their age. Thirty-five per cent of those interviewed were unwilling to accept the risk of cosmetic changes, 38% were unwilling to accept the risk of temporary discomfort and 45% were unwilling to accept the risk of major toxicity. Patients who had previously experienced AEs were more willing to accept the risk of cosmetic changes (83 vs. 58%, P=0.02), temporary discomfort (79 vs. 55%, P=0.02) and major toxicity (83 vs. 44%, P=0.001) compared with those who had not previously experienced AEs. CONCLUSIONS: Many rheumatoid arthritis patients are very concerned about potential drug toxicity. However, risk adversity appeared to be attenuated by past experience with AEs. Our results suggest that certain patients, especially those with milder disease activity, might be reluctant to accept commonly used arthritis medications if they are fully informed of their potential toxicity. PMID- 11934961 TI - Investigation of the chronic pulmonary effects of low-dose oral methotrexate in patients with rheumatoid arthritis: a prospective study incorporating HRCT scanning and pulmonary function tests. AB - OBJECTIVE: Methotrexate has a well-recognized side-effect of acute hypersensitivity pneumonitis. There is concern about whether chronic pulmonary toxicity can occur with methotrexate treatment. Our objective was to compare chest high-resolution computed tomography (HRCT) findings and serial pulmonary function tests in rheumatoid arthritis (RA) patients on methotrexate with findings for a control group of patients with RA who were not being treated with methotrexate. METHODS: Study patients had an initial chest radiograph, full pulmonary function tests and chest HRCT. Pulmonary function tests were then performed regularly over a 2-yr period. RESULTS: Fifty-five RA patients on methotrexate and 73 control patients with RA were enrolled for the study. Mean dose of methotrexate was 10.7 mg/week (S.D. 2.5 mg/week) and mean duration of treatment at entry into the study was 30 (20) months. Twenty per cent of patients with RA treated with methotrexate had pulmonary fibrosis (PF) on initial HRCT compared with 23% in the control group. When the patients with and without PF were compared, there was no statistical difference in the duration (mean difference -4.18 months, P=0.237) or dose (mean difference -0.8 mg/week P=0.52) of methotrexate therapy. Mean changes after 2 yr in forced expiratory volume, forced vital capacity, diffusion capacity for carbon monoxide and residual volumes were not different in the methotrexate group compared with the control group. CONCLUSION: There is no evidence to suggest clinically, from HRCT assessment or serial pulmonary function tests, that low-dose methotrexate is associated with chronic interstitial lung disease. PMID- 11934962 TI - Computerized information-gathering in specialist rheumatology clinics: an initial evaluation of an electronic version of the Short Form 36. AB - OBJECTIVES: Longitudinal outcome data are important for research and are becoming part of routine clinical practice. We assessed an initial version of an electronic Short Form 36 (SF-36), a well-established health assessment questionnaire, in comparison with standard paper forms, in two specialist rheumatology clinics. METHODS: Out-patients (20 with systemic lupus erythematosus and 31 with vasculitis) were randomly selected to complete either paper (n=29) or electronic and paper SF-36 versions (n=51) before and after consultation (paper vs paper comparison). Data were evaluated as the response correlation, internal consistency, missing data, patient satisfaction and preference. RESULTS: There were very good correlations in SF-36 responses (P<0.001) between the paper and electronic forms and the paper and paper forms. Internal reliability coefficients (Cronbach's alpha) showed good internal consistency for all reported responses in either computer or paper forms. There were no missing data in the computerized version but 24% of patients failed to answer all of the paper form questions. Ease of use of the computer version was rated highly by 71% of all the respondents, and 69% would prefer to use the computer version in future. DISCUSSION: Computerized data collection is acceptable to patients and feasible in clinical settings. It provides responses that are at least comparable to those to the paper form, improves data capture and is available immediately. PMID- 11934963 TI - Adjuvant treatment of patients with antineutrophil cytoplasmic antibody associated vasculitis with vitamins E and C reduces superoxide production by neutrophils. AB - OBJECTIVES: Neutrophils when activated generate a respiratory burst which has been implicated in the pathogenesis of primary systemic vasculitis. Neutrophils from patients with vasculitis have a greater respiratory burst than normal healthy donors. The aim of this study was to assess the effects of antioxidant treatment (vitamins E and C) on the generation of a respiratory burst from neutrophils isolated from patients with antineutrophil cytoplasmic antibody (ANCA)-associated vasculitis. METHODS: Neutrophils were isolated from patients with systemic vasculitis and healthy donors. Spontaneous superoxide generation was measured by the reduction of ferricytochrome c. The patients were treated with antioxidants, vitamins E and C, and spontaneous superoxide generation, vitamin C and total antioxidant capacity were measured before and after treatment. RESULTS: The treatment of the patients with antioxidants resulted in a reduction in spontaneous superoxide generation (pre-treatment 8.41+/-0.7 nmol/10(6) cells; post-treatment 5.64+/-0.6 nmol/10(6) cells; P<0.05). There was no significant difference in the superoxide generation from normal controls who did not receive treatment, measured prior to commencement of the study and 10 days later (first reading 4.81+/-0.5 nmol/10(6) cells; second reading 5.32+/-0.4 nmol/10(6) cells; P>0.05). Total antioxidant capacity increased significantly following treatment with vitamins C and E (555.4+/-142 vs. 668.6+/-186 micromol/l trolox equivalent; P=0.01) as did vitamin C concentrations (56.5+/-27 vs. 137.7+/ 64 micromol/l; P=0.002). CONCLUSIONS: In this preliminary study, the treatment of patients with antioxidants, vitamins E and C, reduced neutrophil generation of superoxide and suggests that antioxidants may have an important role as adjuvant therapy. The evidence presented should form the basis of a larger randomized placebo-controlled trial of vitamins E and C as adjuvant therapy in patients with ANCA-associated systemic vasculitis. PMID- 11934964 TI - A randomized, double-blind, placebo-controlled trial of glucosamine sulphate as an analgesic in osteoarthritis of the knee. AB - OBJECTIVES: A randomized, placebo-controlled, double-blind trial of the relative effectiveness of glucosamine sulphate and placebo in managing pain in osteoarthritis (OA) of the knee. METHODS: Eighty patients with OA of the knee were recruited from a rheumatology out-patient clinic and received either glucosamine sulphate 1500 mg daily for 6 months or dummy placebo. The primary outcome measure was patients' global assessment of pain in the affected knee. RESULTS: Area under the curve analysis for the primary outcome measure showed no difference between placebo and glucosamine [mean difference 0.15 mm, 95% confidence interval (CI) -8.78 to 9.07]. The placebo response was 33%. There was a statistically significant difference between groups in knee flexion (mean difference 13 degrees, 95% CI -23.13 to -1.97), but this difference was small and could have been due to measurement error. CONCLUSIONS: As a symptom modifier in OA patients with a wide range of pain severities, glucosamine sulphate was no more effective than placebo. PMID- 11934965 TI - Bioavailable testosterone in men with rheumatoid arthritis-high frequency of hypogonadism. AB - OBJECTIVES: To study bioavailable testosterone (T) in men with rheumatoid arthritis (RA) by determining non-sex hormone-binding globulin (SHBG)-bound T (NST) under standardized conditions and to investigate if NST is related to disease variables. METHODS: Basal serum concentrations of total T, SHBG and luteinizing hormone (LH) were measured in 104 men with RA, and the levels of NST as well as the quotient T/SHBG were calculated. The data were compared with those of 99 age-matched healthy men. The results were analysed separately for the age groups 30-49, 50-59 and 60-69 yr. RESULTS: The RA men had lower NST levels than the healthy men in all age groups. T levels and the T/SHBG ratio were lower only in the age group 50-59 yr. SHBG did not differ significantly. LH was significantly lower in the patients than in the controls. Thirty-three of the 104 patients were considered to have hypogonadism compared with seven of the 99 healthy men. The only clinical variable apart from age that had a significant impact on NST was the Stanford Health Assessment Questionnaire (HAQ). CONCLUSION: Men with RA had lower levels of bioavailable T and a large proportion were considered hypogonadal. The low levels of LH suggested a central origin of the relative hypoandrogenicity. PMID- 11934967 TI - Sjogren's syndrome in an out-patient clinic: classification of patients according to the preliminary European criteria and the proposed modified European criteria. AB - OBJECTIVES: To investigate whether out-patients with a clinical diagnosis of Sjogren's syndrome (SS) satisfied current preliminary European criteria for SS, and to determine the proportion of patients who satisfied the proposed modified European criteria for SS and thus had indication of an autoimmune process. METHODS: Out-patients with a clinical diagnosis of SS registered between 1 January 1999 and 1 November 2000 were included in the study. RESULTS: Of 203 patients with a clinical diagnosis of SS, 116 (57.1%) satisfied the current European criteria and 83 (40.9%) satisfied the proposed modified criteria. CONCLUSIONS: Sicca symptoms and signs may have a variety of causes. In our study only 40.9% of the patients with a clinical diagnosis of SS satisfied the proposed modified European criteria and had evidence of SS as an autoimmune condition. Our findings indicate that for patient populations with an established diagnosis of SS according to the preliminary European criteria, approximately one-third will lose the diagnosis according to the modified criteria. PMID- 11934966 TI - Characteristics and outcome of 49 patients with symptomatic cryoglobulinaemia. AB - OBJECTIVE: To describe a population of patients with symptomatic cryoglobulinaemia, comparing manifestations and outcome as a function of hepatitis C virus (HCV) status. PATIENTS AND METHODS: A retrospective study on 179 patients who tested positive for cryoglobulins, seen between 1978 and 1998 in an internal medicine department. RESULTS: Among 179 cryoglobulin-positive patients, only 49 (18 men, 31 women; mean age 59.96+/-12 yr) had clinical manifestations attributable to cryoglobulinaemia. Thirty-three had HCV infection, 20 had systemic autoimmune diseases, two had haematological diseases, one had human immunodeficiency virus and HCV co-infection, one had HCV and HBV co infection and six had essential mixed cryoglobulinaemia. The clinical manifestations and cryoglobulin levels in HCV(+) and HCV(-) patients did not differ significantly. Only arthralgias and elevated transaminases were significantly more frequent in HCV(+) patients (P<0.02 and <0.05, respectively). Five-year survival rates were comparable for HCV(+) and HCV(-) patients. Eight patients died (six HCV(+), two HCV(-)), with a median time between diagnosis and death of 38.7 months. CONCLUSION: Clinical manifestations of cryoglobulinaemia, except arthralgias, were comparable for HCV(+) and HCV(-) patients. When systemic manifestations are present, the prognosis is poor despite intensive or prolonged therapy. PMID- 11934968 TI - The contribution of HLA-DQB1 coding and QBP promoter alleles to anti-Ro alone autoantibody response in systemic lupus erythematosus. AB - OBJECTIVE: To analyse the influence of HLA-DR, DQ and corresponding DQA1 and DQB1 promoter alleles (QAP and QBP) on the anti-Ro alone autoantibody response in systemic lupus erythematosus (SLE). METHODS: Sixty-five unrelated anti-La antibody-negative SLE patients, 37 of them with and 28 without anti-Ro antibodies, were included. Anti-Ro antibodies were determined by both counter immunoelectrophoresis and enzyme-linked immunosorbent assay. Seventy-four healthy individuals were selected as controls. The patients and controls were analysed for HLA-DRB1, QAP, DQA1, QBP and DQB1 alleles by DNA typing. The allelic frequencies of anti-Ro alone-positive and anti-Ro-negative SLE patients and healthy controls were compared using the chi(2) test or Fisher's exact test as appropriate. RESULTS: The DQB1*0202 allele showed a significant positive correlation with anti-Ro alone antibodies [odds ratio (OR)=16.949, P=0.0015, corrected P=0.018], while the QBP5.11 allele and the combination of DQB1*0301 and its promoter QBP3.1 were under-represented in anti-Ro-alone-positive SLE patients (P=0.01, corrected P=0.048 and corrected P=0.048 respectively). CONCLUSIONS: The above-mentioned alleles may contribute to the presence or absence of anti-Ro alone autoantibodies in SLE patients. PMID- 11934969 TI - Campylobacter-triggered reactive arthritis: a population-based study. AB - OBJECTIVE: To study the incidence and clinical picture of Campylobacter associated reactive arthritis (ReA) and other reactive musculoskeletal symptoms in the population. METHODS: A questionnaire on enteric and extraintestinal, including specifically musculoskeletal, symptoms was sent to 870 consecutive patients with Campylobacter-positive stool culture and 1440 matched controls. Analysis of self-reported musculoskeletal symptoms with clinical examination was performed. RESULTS: Forty-five of the patients (7%) had ReA and eight (1%) had reactive tendinitis, enthesopathy or bursitis. No child had ReA. The arthritis was oligo- or polyarticular, and, in most cases, mild. HLA-B27 was positive in 14% of ReA patients. Of the 45 ReA patients, 37 had C. jejuni and 8 had C. coli infection. No controls had ReA. CONCLUSION: ReA is common following Campylobacter infection, with an annual incidence of 4.3 per 100000. At the population level, acute ReA is mild, more frequent in adults, and not associated with HLA-B27. Besides C. jejuni, C. coli can trigger ReA. PMID- 11934970 TI - Dendritic cells in rheumatoid synovial membrane after total removal of the hyaline articular cartilage. AB - OBJECTIVE: To investigate the effect of total removal of the hyaline articular cartilage on dendritic cells in synovial membrane in rheumatoid arthritis (RA) or ankylosing spondylitis (AS). PATIENTS AND METHODS: Immunohistochemical staining for two dendritic cell markers, CD35 and RFD1, was carried out on synovial membrane specimens from arthritis patients undergoing primary (n=10) or revision (n=8) total hip replacement (THR). The results are expressed as the number (mean+/-standard deviation) of positive cells per 1000 total cells. RESULTS: CD35 (112+/-9) and RFD1-(27+/-5) positive cells were found in all primary RA synovial membrane, while only two out of eight synovial membrane samples from revision THR contained CD35-positive follicular dendritic cells (nine and 12 cells), and no revision samples contained any RFD1-positive interdigitating dendritic cells. CONCLUSION: Removal of the hyaline articular cartilage reduces the infiltration and functional differentiation of dendritic cells in synovial membrane. Our findings suggest that the antigen driving chronic arthritis/synovitis is contained in the hyaline articular cartilage. PMID- 11934971 TI - Digital vascular response to topical glyceryl trinitrate, as measured by laser Doppler imaging, in primary Raynaud's phenomenon and systemic sclerosis. AB - OBJECTIVE: To investigate digital microvascular responses to topical glyceryl trinitrate (GTN) in patients with primary Raynaud's phenomenon (PRP), limited cutaneous systemic sclerosis (LCSSc) and healthy control subjects, using laser Doppler imaging. METHODS: Ten patients with PRP, 13 with LCSSc and 10 control subjects were studied. Baseline skin microvascular blood flow of the dorsum of the index, middle and ring fingers of the non-dominant hand was measured using scanning laser Doppler imaging. After the initial image, 2% GTN ointment was rubbed on the dorsum of one finger for 1 min; placebo ointment was rubbed on the dorsum of a second finger for 1 min, and the third finger remained untreated. Further laser Doppler scanning of these three fingers was conducted immediately, 10 and 20 min after ointment application. RESULTS: There was increased blood flow response to placebo compared with no treatment (P<0.001) and to GTN compared with placebo (P=0.004). The change in blood flow over time differed significantly between placebo and GTN (P<0.001), but not between placebo and no ointment application: blood flow increased with GTN and decreased with placebo/no treatment at 10 and 20 min. There were no differences in initial baseline blood flow or response between the subject groups. CONCLUSIONS: An exogenous supply of nitric oxide by topical GTN ointment causes local endothelial-independent vasodilatory responses in PRP, LCSSc patients and control subjects. As well as demonstrating the effectiveness of topical GTN in patients with PRP and LCSSc, this study illustrates the ability of laser Doppler imaging to quantify local vasodilatory effects. PMID- 11934974 TI - Antinuclear factor. PMID- 11934972 TI - The role of TNF-alpha in the pathogenesis of inflammation and joint destruction in rheumatoid arthritis (RA): a study using a human RA/SCID mouse chimera. AB - OBJECTIVE: In order to elucidate which cytokine preferentially stimulates the synovium in patients with rheumatoid arthritis (RA), we investigated the roles of tumour necrosis factor alpha (TNF-alpha) and interleukin 6 (IL-6) using SCID mice engrafted with human RA tissue (SCID-HuRAg). METHODS: The SCID-HuRAg mice were prepared according to our previously described method. First, SCID-HuRAg mice were treated with chimeric anti-TNF-alpha monoclonal antibody (mAb, 100 microg/mouse) and histological changes were examined 4 weeks after the initial treatment. Secondly, a total of 100 microg of recombinant TNF-alpha or IL-6 (0.6 microg/h) was administered daily to mice using an osmium pump. The histological changes and serum cytokine levels were examined 4 weeks after the initial administration. Human immunoglobulin G (IgG) was administered to mice as a control. RESULTS: Synovial inflammatory cells were significantly decreased after the anti-TNF-alpha mAb treatment; conversely, the degree of synovial inflammation was significantly exacerbated by TNF-alpha administration. The levels of both IL 6 and TNF-alpha in sera were significantly increased by recombinant TNF-alpha administration, while TNF-alpha levels were unchanged by IL-6 administration. This suggests that TNF-alpha controls IL-6 production. Despite the profound changes in inflammation, we found no effects on bone and no articular cartilage damage was produced by TNF-alpha. CONCLUSION: This study provides strong evidence that TNF-alpha is a key molecule in the control of the inflammatory changes that occur in the RA synovium. In addition, TNF-alpha regulates IL-6 production. However, other inflammatory pathways independent of TNF-alpha may contribute to the bone and cartilage damage seen in RA. PMID- 11934973 TI - Activated caspase 3 and cleaved poly(ADP-ribose)polymerase in salivary epithelium suggest a pathogenetic mechanism for Sjogren's syndrome. AB - OBJECTIVE: Apoptosis is an organized energy-dependent process of cellular self destruction carried out by proteolytic enzymes such as the caspases. These enzymes may play a role in epithelial cell apoptosis in Sjogren's syndrome (SS). A classical caspase substrate is poly(ADP-ribose)polymerase (PARP), a DNA repair enzyme. To elucidate the molecular mechanisms responsible for salivary gland dysfunction in SS, we studied the expression of caspase and PARP in SS salivary gland biopsies. METHODS: The presence of activated caspases (caspases 3 and 9) and cleaved PARP (85 kDa) in SS biopsies was demonstrated by immunohistochemistry using specific polyclonal antibodies. RESULTS: Initial studies performed with an antibody reagent that recognizes both active and inactive forms of caspase 3 identified this enzyme in SS salivary ductal and acinar cells. Activated caspase 3 and cleaved PARP were strongly expressed in ductal and acinar cells in SS salivary glands (13/15). Ductal and acinar cells from normal salivary glands (n=5) stained with less intensity compared with SS tissue. Staining for activated caspase 9 was negative in all samples. Likewise, infiltrating lymphocytes were negative for caspase 3, caspase 9 and cleaved PARP. CONCLUSION: This study shows that caspase 3 is important in the salivary dysfunction of SS, while caspase 9 appears not to be involved. PMID- 11934975 TI - Alendronate in the treatment of avascular necrosis of the hip. PMID- 11934976 TI - Training in musculoskeletal sonography: report from the first BSR course. PMID- 11934977 TI - Anti-tumour necrosis factor treatment with infliximab in a case of giant cell arteritis resistant to steroid and immunosuppressive drugs. PMID- 11934978 TI - Idiopathic hypereosinophilic syndrome in a patient with long-standing rheumatoid arthritis: a case report. PMID- 11934979 TI - The relationship between soft tissue swelling, joint space narrowing and erosive damage in hand X-rays of patients with rheumatoid arthritis. PMID- 11934980 TI - Inappropriate serum levels of IGF-I and IGFBP-3 in patients with rheumatoid arthritis. PMID- 11934981 TI - Ibuprofen-induced aseptic meningoencephalitis. PMID- 11934982 TI - Haemorrhagic colitis due to Escherichia coli O103:H2 associated with infliximab therapy in a patient with rheumatoid arthritis. PMID- 11934983 TI - Rheumatology day care and the impact of biological therapies-the Belfast experience. PMID- 11934987 TI - Transcription control by E1A and MAP kinase pathway via Sur2 mediator subunit. AB - Sur2 is a metazoan Mediator subunit that interacts with the adenovirus E1A protein and functions in a mitogen-activated protein kinase pathway required for vulva development in Caenorhabditis elegans. We generated sur2-/- embryonic stem cells to analyze its function as a mammalian Mediator component. Our results show that Sur2 forms a subcomplex of the Mediator with two other subunits, TRAP/Med100 and 95. Knock-out of Sur2 prevents activation by E1A-CR3 and the mitogen activated protein kinase-regulated ETS transcription factor Elk-1, but not by multiple other transcription factors. These results imply that specific activation domains stimulate transcription by binding to distinct Mediator subunits. Activation by E1A and Elk-1 requires recruitment of Mediator to a promoter by binding to its Sur2 subunit. PMID- 11934988 TI - Genomic instability in mice lacking histone H2AX. AB - Higher order chromatin structure presents a barrier to the recognition and repair of DNA damage. Double-strand breaks (DSBs) induce histone H2AX phosphorylation, which is associated with the recruitment of repair factors to damaged DNA. To help clarify the physiological role of H2AX, we targeted H2AX in mice. Although H2AX is not essential for irradiation-induced cell-cycle checkpoints, H2AX-/- mice were radiation sensitive, growth retarded, and immune deficient, and mutant males were infertile. These pleiotropic phenotypes were associated with chromosomal instability, repair defects, and impaired recruitment of Nbs1, 53bp1, and Brca1, but not Rad51, to irradiation-induced foci. Thus, H2AX is critical for facilitating the assembly of specific DNA-repair complexes on damaged DNA. PMID- 11934989 TI - Observations of comet 19P/Borrelly by the miniature integrated camera and spectrometer aboard Deep Space 1. AB - The nucleus of the Jupiter-family comet 19P/Borrelly was closely observed by the Miniature Integrated Camera and Spectrometer aboard the Deep Space 1 spacecraft on 22 September 2001. The 8-kilometer-long body is highly variegated on a scale of 200 meters, exhibiting large albedo variations (0.01 to 0.03) and complex geologic relationships. Short-wavelength infrared spectra (1.3 to 2.6 micrometers) show a slope toward the red and a hot, dry surface ( or =40 years and with stroke, AMI, or peripheral arterial disease (PAD) were identified from administrative data of UnitedHealthcare plans during 1995-1998. Stroke, AMI, and PAD cohorts were identified within a commercial insurance sample and a Medicare sample. Cumulative occurrences of subsequent stroke, AMI, or vascular death were estimated by survival analysis. RESULTS: In the stroke commercial cohort (n=1631; mean age, 62.1 years), cumulative occurrence of subsequent events was 4.2%, 6.5%, 9.8%, and 11.8% at 0.5, 1, 2, and 3 years, respectively; cumulative secondary event occurrence in the AMI commercial cohort (n=6458; mean age, 56.0 years) was 3.5%, 4.8%, 7.3%, and 8.5% and in the PAD commercial cohort (n=5813; mean age, 59.2 years) was 1.5%, 2.8%, 4.8%, and 6.5%, respectively. Cumulative secondary event occurrences were even higher in stroke (n=1518; mean age, 79.5 years), AMI (n=2197; mean age, 76.2 years), and PAD (n=5033; mean age, 76.6 years) cohorts of the Medicare sample: 18.1%, 17.0%, and 8.7%, respectively, at 3 years. More than 75% of each stroke cohort's secondary events were strokes; more than 75% of each AMI cohort's secondary events were AMIs. Of the PAD cohorts' secondary events, 27% to 39% were strokes, 48% to 57% were AMIs, and 13% to 16% were vascular deaths. CONCLUSIONS: Among these managed care enrollees with existing atherosclerotic vascular disease, subsequent ischemic events represent a significant symptomatic disease burden. Given these findings, it is very important to determine whether secondary prevention strategies are being effectively used to manage patients with diagnosed atherosclerosis. PMID- 11935035 TI - Alcohol consumption and risk of ischemic stroke: The Framingham Study. AB - BACKGROUND AND PURPOSE: Stroke is a major cause of death in the United States. The association between alcohol consumption and ischemic stroke (IS) remains controversial. METHODS: We used data collected on participants in the Framingham Study to assess the association between total alcohol intake and type of alcoholic beverage and development of IS, overall and according to age. RESULTS: A total of 196 men and 245 women developed IS during three 10-year follow-up periods. In the categories of never drinkers, drinkers of 0.1 to 11, 12 to 23, and > or =24 g/d of ethanol (a "typical drink" is approximately 12 g of ethanol), and former drinkers of 0.1 to 11 and > or =12 g/d, crude incidence rates of IS were 6.5, 5.9, 4.9, 5.0, 6.7, and 17.8 cases per 1000 person-years, respectively, for men and 5.9, 4.1, 4.1, 4.3, 8.3, and 7.1, respectively, for women. Overall, compared with never drinkers in a multivariate Cox regression, current alcohol consumption was not related significantly to IS in either sex. Former drinking of > or =12 g/d of alcohol was associated with a 2.4 times higher risk of IS among men but not among women. When stratified by age, alcohol intake was associated with lower risk of IS among subjects aged 60 to 69 years. In beverage-specific analysis, only wine consumption was related to a decreased risk of IS. CONCLUSIONS: Our data showed no significant association between total alcohol and IS overall but showed a protective effect of alcohol among subjects aged 60 to 69 years. PMID- 11935037 TI - Angioarchitectural factors present in brain arteriovenous malformations associated with hemorrhagic presentation. AB - BACKGROUND AND PURPOSE: Associations between clinical presentation of brain arteriovenous malformations (AVMs) and their angioarchitecture have been described. This study aims to identify significant factors related to the initial hemorrhagic event through multivariate statistical methodology. METHODS: The authors studied the initial clinical presentation of 390 consecutive patients with brain AVMs at the University of Toronto Vascular Malformation Study Group. Angiographic features present at that time, such as location, size, and blood supply, were recorded following a standard protocol and associated, through multivariate analysis techniques, with type of presentation. RESULTS: Patients had hemorrhagic presentation in 146 cases (37.4%). Hemorrhage was the initial presentation in 59.5% of the deep-seated AVMs (odds ratio [OR]=3.26; 95% CI=1.15 to 9.2; P=0.03). A single draining vein was associated with bleeding at presentation in 57.6% AVMs (OR=1.78; 95% CI=1.12 to 2.82; P=0.01), and 72.8% of the patients with venous ectasia had bleeding as initial evidence (OR=3.9; 95% CI=1.63 to 9.28; P=0.002). Hemorrhage was the initial presentation in 47.6% (111/233) of AVMs <3 cm, 22.5% (32/142) in sizes between 3 and 6 cm, and 20% in malformations >6 cm (3/15), but these differences were not significant in multivariate analyses. CONCLUSIONS: For initial hemorrhagic presentation, a small number of draining veins, deep location, and the presence of venous ectasias were significant associated factors. In contrast with many previous reports, AVM size was not associated with hemorrhage at presentation in adjusted analyses. PMID- 11935036 TI - Stroke risk factor profiles in African American women: an interim report from the African-American Antiplatelet Stroke Prevention Study. AB - BACKGROUND AND PURPOSE: If sex differences in stroke risk factor profiles exist among African Americans in the United States, prevention strategies will need to reflect those differences. African Americans and women have been underrepresented in stroke prevention studies. The purpose of this study was to determine whether medical and lifestyle factors differ among women and men who have enrolled in the African-American Antiplatelet Stroke Prevention Study (AAASPS). METHODS: We performed a planned exploratory analysis of differences in baseline characteristics and risk factors between women and men enrolled in AAASPS, a double-blind, randomized, multicenter, controlled trial. Frequencies of vascular risk factors and related conditions, medical therapies, stroke subtypes, and vascular territories were compared between women and men by 1-way ANOVA and Fisher's exact test where appropriate. RESULTS: A total of 1087 African American patients (574 women, 513 men) enrolled between December 1995 and June 1999. Women had higher rates of hypertension, diabetes, family history of stroke, and no reported leisure exercise. Men had higher rates of smoking and heavy alcohol use. Few differences were noted in proportions of stroke subtype or proportions receiving preventive therapy. CONCLUSIONS: AAASPS represents the largest enrollment of African American women in a recurrent stroke prevention study. Our data suggest that African American women in a clinical trial differ from men in the frequency of key vascular risk factors. Although limited, these data provide an important first characterization of sex differences in African Americans with stroke. PMID- 11935038 TI - Hypertensive pontine microhemorrhage. AB - BACKGROUND AND PURPOSE: This study investigated whether the topography of hypertensive pontine microhemorrhages (hPMHs) resembles that of larger primary pontine hemorrhages. METHODS: Sixty-nine consecutive patients with small-vessel disease underwent imaging with gradient-echo MRI, and 27 patients with hPMH were detected. Lesion size and location along the rostrocaudal (longitudinal), lateral (coronal), and anteroposterior (sagittal) axes were determined. RESULTS: A total of 52 hPMHs were identified in the 27 patients (mean, 1.93+/-2.4 per patient). The lesions showed a nonrandom distribution, with a propensity to occur in the middle pons in the rostrocaudal axis, posterior half of the basis pontis in the anteroposterior axis, and central subdivision within the lateral axis. The area of hPMH ranged from 1.3 to 19.0 mm2 (mean, 5.06+/-3.72 mm2). The size of hPMH did not vary as a function of lesion location. CONCLUSIONS: Previous studies reported that primary pontine hemorrhages tend to occur in the middle pons and at the junction of basis pontis and tegmentum. Therefore, topographical correspondences between large and small pontine hemorrhages may provide evidence that the 2 lesions share some etiological basis. Further investigation may determine whether hPMHs portend future symptomatic primary pontine hemorrhages. PMID- 11935039 TI - Risk factors for progression of aortic atheroma in stroke and transient ischemic attack patients. AB - BACKGROUND AND PURPOSE: Aortic atheroma is an independent risk factor for stroke and undergoes temporal progression. Clinical and risk factor associations of such progression are unknown. Hyperhomocysteinemia has been linked with atherosclerosis, including that in the cerebral vasculature. This study investigated associations between elevated homocysteine levels and other stroke vascular risk factors and the risk of aortic atheroma progression in patients with cerebrovascular disease. METHODS: Fifty-seven stroke and 21 transient ischemic attack patients underwent multiplanar transesophageal echocardiograms within 1 month of symptom onset and again at 9 months. Aortic atheroma was graded and stratified by use of existing criteria. Stroke risk factors; use of anticoagulant, antiplatelet, and hypolipidemic drugs; and clinical and etiological subtypes of stroke were recorded and compared in patients stratified for the presence or absence of aortic atheroma progression. RESULTS: Of the 78, 29 (37%) progressed, 32 (41%) remained unchanged, and 17 (22%) regressed. Progression was most marked at the aortic arch (P=0.005), followed by the ascending segment (P<0.04). In nearly two thirds of the patients in whom aortic atheroma remained unchanged over 9 months, no atheroma was evident on baseline transesophageal echocardiogram. Only homocysteine levels > or =14.0 micromol/L (P=0.02), total anterior cerebral infarct (P=0.02), and large-artery atherosclerosis (P=0.005) significantly correlated with progression. CONCLUSIONS: Among vascular risk factors, elevated homocysteine levels are associated with aortic atheroma progression. Stroke and transient ischemic attack patients with aortic atheroma should undergo assessment of homocysteine levels, which, if elevated, may be treated with vitamins in an effort to arrest aortic atheroma progression. PMID- 11935040 TI - Increased circulating immune complexes in acute stroke: the triggering role of Chlamydia pneumoniae and cytomegalovirus. AB - BACKGROUND AND PURPOSE: The mechanisms of immune reaction involved in the pathogenesis and clinical course of acute vascular incidents are still not completely understood. The aim of this study was to examine the presence of immune complexes (IC) in the acute stroke setting and the first month thereafter and to characterize IC by analyzing the contents of chlamydial lipopolysaccharide and anti-cytomegalovirus (CMV) antibodies in IC. METHODS: Serum concentration of IC was investigated in 179 stroke patients, 122 "old" controls and 112 "young" controls, by the precipitation method. The presence of chlamydial lipopolisaccharyde and anti-CMV antibodies was investigated in some IC preparations by the ELISA method after earlier dissociation of IC into components by high pH treatment. RESULTS: Significantly increased serum IC concentration in stroke patients was noticed. Increased serum IC concentration was revealed as an independent strong stroke risk factor and was connected with significantly worse neurological status and increased 30-day mortality rate. A significantly larger proportion of stroke patients than controls had Chlamydia pneumoniae antigen and anti-CMV antibodies in IC. CONCLUSIONS: This study provides the first evidence of an association between increased serum level of IC and the clinical course of cerebral ischemia and identifies a potentially important association of C pneumoniae and CMV-specific IC with stroke incidence. PMID- 11935041 TI - Dissecting aneurysms of intracranial carotid circulation. AB - BACKGROUND AND PURPOSE: Clinical features of nontraumatic dissecting aneurysms of intracranial carotid circulation remain unclear because investigation of this disease has been limited to case reports. The aim of this study was to investigate the clinical features of this disease through the use of cooperatively collected cases. METHODS: The cases diagnosed as dissecting aneurysms of intracranial carotid circulation on the basis of clinical signs and neuroradiological findings in 46 stroke centers from 1995 through 1999 were collected, and their clinical features were analyzed. RESULTS: Forty-nine cases of dissecting aneurysms of intracranial carotid circulation were collected. Thirty-two patients presented with subarachnoid hemorrhage (SAH), and 17 presented with cerebral ischemia. The ratio of this disease to all intracranial dissecting aneurysms treated in the same institutes for the same period was 19.1%, and the ratio of SAH resulting from this disease to SAH of unverified origin treated in the same institutes for the same period was 6.2%. The predominant site of lesion was the internal carotid artery in 18 of 32 patients (56%) with SAH and the anterior cerebral artery in 13 of 17 patients (76%) with cerebral ischemia. The predominant angiographic findings were that stenosis with dilatation occurred in 20 of 32 patients (63%) with SAH and stenosis without dilatation was seen in 11 of 17 patients (65%) with cerebral ischemia. Poor prognosis was seen in 21 of 32 patients (66%) with SAH, which was due largely to rebleeding seen preoperatively, during operation, and even postoperatively when clipping or wrapping of the aneurysmal bulge was performed. CONCLUSIONS: Nontraumatic dissecting aneurysm of intracranial carotid circulation is not as rare as expected. It seems to be one of the important causes of SAH of unverified origin. PMID- 11935042 TI - Physician knowledge and practices in the evaluation of coagulopathies in stroke patients. AB - BACKGROUND AND PURPOSE: Coagulopathies are a rare cause of ischemic stroke. Prior studies demonstrate that current physician test-ordering practices for the evaluation of these conditions in patients with ischemic stroke is not optimal. We sought to determine neurologists' views regarding their use of specialized coagulation testing to better understand the possible reasons for these practices. METHODS: A survey with multiple-choice and open-ended questions regarding knowledge of and approaches to the evaluation of coagulopathies was sent to a convenience sample of 79 neurologists (26 academic neurology faculty, 24 residents/fellows, and 29 community-based practitioners). RESULTS: Fifty-nine (75%) surveys were completed (response rates: faculty 73%, residents/fellows 88%, and community-based practice 66%). Specialized coagulation tests were reported to infrequently influence stroke patient management (<25% of the time or never for 95% of respondents). Factors reported to increase test-ordering included young patient age (76%), history of thrombosis (46%), history of miscarriages (36%), and having few traditional stroke risk factors (35%). Most (88%) indicated they would order specialized coagulation tests for a hypothetical young patient with no known stroke risk factors. In contrast, only 14% would obtain the tests for a patient having traditional stroke risk factors, and none would order the tests for a stroke patient with atrial fibrillation. CONCLUSIONS: Physician-reported practices for obtaining specialized coagulation tests differ from those found in observational studies in which more indiscriminate test ordering was observed. Closing knowledge gaps and improving application of physician's current knowledge to their test-ordering practices could help to optimize diagnostic testing for coagulopathies in patients with ischemic stroke. PMID- 11935043 TI - Is the association of National Institutes of Health Stroke Scale scores and acute magnetic resonance imaging stroke volume equal for patients with right- and left hemisphere ischemic stroke? AB - BACKGROUND AND PURPOSE: The National Institutes of Health Stroke Scale (NIHSS) is an established measure of neurological impairment; however, it can award more points for tests of presumed left-hemisphere function, such as language, than for tests of right-hemisphere function, such as neglect. This difference may be important if a low NIHSS score is used to exclude patients with right-hemisphere stroke from clinical trials or established treatments. The aim of this study was to investigate whether the relationship between acute NIHSS score and acute stroke volume as determined by acute diffusion- and perfusion-weighted MRI (DWI and PWI) differs between right- and left-sided stroke. METHODS: This was a retrospective study of 153 patients with acute stroke seen at Beth Israel Deaconess Medical Center between January 1995 and March 2000 who underwent an MRI examination and NIHSS within 24 hours of stroke onset. NIHSS score was recorded prospectively by the admitting stroke fellow at the time of acute presentation, immediately preceding imaging. Computerized volumetric analysis of the MRI lesions was performed by investigators blinded to clinical data. RESULTS: There were significant correlations between the acute NIHSS scores and acute DWI lesion volumes (r=0.48 right, r=0.58 left) and between acute NIHSS scores and perfusion weight imaging hypoperfusion volumes (r=0.62 right, r=0.60 left). For patients with NIHSS scores of 0 to 5, the DWI volume of right cerebral lesions was greater than that of left-sided lesions (mean volume, 8.8 versus 3.2 cm3; P=0.04). Among patients with DWI lesions larger than the median volume (9 cm3), 8 of 37 with right-sided stroke had an NIHSS score of 0 to 5 compared with 1 of 39 patients with left-sided stroke (P=0.01). Multiple linear regression analysis revealed a significantly lower acute NIHSS on the right compared with the left side when adjusted for stroke volume on chronic T2 imaging (P=0.03). CONCLUSIONS: Patients with right-sided stroke may have a low NIHSS score despite substantial DWI lesion volume. Acute imaging information, such as that available with multimodal MRI, may be useful to identify patients for inclusion in acute stroke protocols when there is clinical uncertainty about eligibility. Prospective evaluation of criteria incorporating acute imaging data is required. PMID- 11935044 TI - CT angiography with whole brain perfused blood volume imaging: added clinical value in the assessment of acute stroke. AB - BACKGROUND AND PURPOSE: In CT angiographic and perfusion imaging (CTA/CTP), rapid CT scanning is performed during the brief steady state administration of a contrast bolus, creating both vascular phase images of the major intracranial vessels and perfused blood volume-weighted parenchymal phase images of the entire brain. We assessed the added clinical value of the data provided by CTA/CTP over that of clinical examination and noncontrast CT (NCCT) alone. METHODS: NCCT and CTA/CTP imaging was performed in 40 patients presenting with an acute stroke. Short clinical vignettes were retrospectively prepared. After concurrent review of the vignettes and NCCT, a stroke neurologist rated infarct location, vascular territory, vessel(s) occluded, and Trial of Org 10172 in Acute Stroke Treatment (TOAST) and Oxfordshire Community Stroke Project classifications. The ratings were repeated after serial review of each of the CTA/CTP components: (1) axial CTA source images; (2) CTP whole brain blood volume-weighted source images; and (3) maximum-intensity projection 3-dimensional reformatted images. The sequential ratings for each case were compared with the final discharge assessment. RESULTS: Compared with the initial review after NCCT, CTA/CTP improved the overall accuracy of infarct localization (P<0.001), vascular territory determination (P=0.003), vessel occlusion identification (P<0.001), TOAST classification (P=0.039), and Oxfordshire Community Stroke Project classification (P<0.001) by 40%, 28%, 38%, 18%, and 32%, respectively. CONCLUSIONS: Admission CTA/CTP imaging significantly improves accuracy, over that of initial clinical assessment and NCCT imaging alone, in the determination of infarct localization, site of vascular occlusion, and Oxfordshire classification in acute stroke patients. PMID- 11935045 TI - Detection of deoxygenation-related signal change in acute ischemic stroke patients by T2*-weighted magnetic resonance imaging. AB - BACKGROUND AND PURPOSE: Acute decreases in the MR T2*-weighted signal have been reported in experimental models of middle cerebral artery occlusion. This has been attributed to blood deoxygenation in association with an increased brain oxygen extraction fraction. The aim of this study was to detect this signal by susceptibility-weighted MR imaging in acute ischemic stroke patients. METHODS: Dynamic susceptibility contrast-enhanced MR (DSC-MR) imaging was performed within 4 hours of stroke onset in 6 patients with unilateral cerebral artery occlusion (middle cerebral artery, n=5; internal carotid artery, n=1). Cerebral blood volume was estimated on a pixel-by-pixel basis. DSC-MR images taken before arrival of the contrast medium were examined visually to identify hypointense areas. Bilateral regions of interest were set in the middle cerebral artery territory for comparison of the mean signal intensity. A semilogarithmic plot of signal intensity versus cerebral blood volume for every pixel in the region of interest was also analyzed. RESULTS: The side on which the hypointense area was seen was significantly correlated with the side of arterial occlusion. The mean signal intensity was significantly smaller on the affected side than on the contralateral side. The semilogarithmic plot of signal intensity versus cerebral blood volume indicated greater deoxyhemoglobin concentrations for the ipsilateral than for the contralateral region of interest. CONCLUSIONS: DSC-MR images allow detection of hypointensity in the affected cerebral hemisphere in acute ischemic stroke patients. Such hypointensity may indicate increased oxygen extraction fraction (misery perfusion) and may provide information valuable to patient care. PMID- 11935046 TI - Cerebral perfusion and cerebrovascular reactivity are reduced in white matter hyperintensities. AB - BACKGROUND AND PURPOSE: There is growing evidence that white matter hyperintensities (WMH) should not be considered as benign age-dependent changes on MR images but indicate pathological changes with clinical consequences. Previous studies comparing subjects with WMH to normal controls have reported global reductions in cerebral blood flow (CBF) and cerebral vascular reactivity. In this study, we examined localized hemodynamic status to compare WMH to normal appearing white matter (NAWM). METHODS: A group of 21 normal 85-year-old subjects were studied using dynamic contrast-enhanced MRI together with administration of acetazolamide. From a combination of anatomic images with different signal weighting, regions of interest were generated corresponding to gray and white matter and WMH. Localized measurements of CBF and cerebral blood volume (CBV) and mean transit time were obtained directly within WMH and NAWM. RESULTS: When comparing WMH to NAWM, measurements showed significantly lower CBF (P=0.004) and longer mean transit time (P< 0.001) in WMH but no significant difference in CBV (P=0.846). The increases in CBF and CBV induced by acetazolamide were significantly smaller in WMH than in NAWM (P=0.026, P<0.001). CONCLUSION: These results show that a change in the hemodynamic status is present within the WMH, making these areas more likely to be exposed to transient ischemia inducing myelin rarefaction. In the future, MRI may be used to examine the effect of therapeutic strategies designed to prevent or normalize vascular changes. PMID- 11935047 TI - Computed tomography angiography for the evaluation of carotid atherosclerotic plaque: correlation with histopathology of endarterectomy specimens. AB - BACKGROUND AND PURPOSE: The goal of this study was to determine the efficacy of CT angiography for the characterization of plaque morphology and composition in carotid atherosclerotic disease. METHODS: Fifty-five patients undergoing carotid endarterectomy were imaged preoperatively with single-slice spiral CT angiography. One hundred sixty-five endarterectomy sections were examined histologically at selected levels through the distal common and proximal internal carotid arteries. Plaque density was measured (in Hounsfield units) on axial CT sections, and the presence or absence of ulceration was noted. These observations were compared with the histological findings at corresponding levels. Data were analyzed with 2-sample t tests and 1-way analysis of variance (ANOVA). RESULTS: ANOVA testing showed a statistically significant decrease in CT attenuation values with increasing plaque lipid but with a very high standard deviation of values. No other histological factor showed a statistically significant link with CT attenuation. Plaque ulceration was detected by CT with a sensitivity of 60% and a specificity of 74%. CONCLUSIONS: Analysis of plaque attenuation with single slice spiral CT does not give useful information concerning plaque composition. The predictive value of CT for the detection of plaque ulceration was moderate. Single-slice CT angiography is insufficiently robust to be a useful tool for the characterization of carotid plaque composition and morphology. PMID- 11935048 TI - Inflammation-mediated damage in progressing lacunar infarctions: a potential therapeutic target. AB - BACKGROUND AND PURPOSE: The mechanisms underlying neurological deterioration in patients with lacunar infarction are not completely understood. In this study, we sought to investigate the role of proinflammatory molecules in the early worsening and outcome of acute lacunar stroke. METHODS: We performed a secondary analysis of 113 consecutive patients with lacunar infarction included within the first 24 hours of the onset of symptoms in a previous study aimed at investigating clinical and biochemical factors of early neurological deterioration (END). END was defined as a fall of > or =1 points in the motor items of Canadian Stroke Scale between inclusion and 48 hours. Poor outcome at 3 months was considered death or Barthel Index <85. Interleukin-6 (IL-6), tumor necrosis factor-alpha (TNF-alpha), and intercellular adhesion molecule-1 (ICAM-1) were determined by enzyme-linked immunoabsorbent assay in blood samples obtained on admission. RESULTS: END was recorded in 27 patients (23.9%); poor outcome was noted in 26 (23%). Median (quartiles) concentrations in plasma of TNF-alpha [16.5 pg/mL (13.7 and 21.2 pg/mL) versus 7.5 pg/mL (6.2 and 9.0 pg/mL)], IL-6 [28.8 pg/mL (22.5 and 35.7 pg/mL) versus 11.5 pg/mL (8.5 and 16.2 pg/mL)], and ICAM-1 [285 pg/mL (219 and 315 pg/mL) versus 158 pg/mL (137 and 187 pg/mL)] were significantly higher in patients who had END than in those with nonprogressing strokes (P< 0.001). Significant differences were also observed between patients with poor and good outcome at 3 months. Logistic regression analysis after adjustment for potential confounders showed that TNF-alpha >14 pg/mL and ICAM-1 >208 pg/mL were independently associated with both END (OR, 511; 95% CI, 17 to 4937; P<0.001; and OR, 315; 95% CI, 17 to 5748; P< 0.001, respectively) and poor outcome at 3 months (OR, 3.0; 95% CI, 1.0 to 8.5; P=0.042; and OR, 4.2; 95% CI, 1.3 to 13.6; P<0.015, respectively). CONCLUSIONS: High concentrations of inflammatory markers in blood are associated with END and poor functional outcome in lacunar infarctions. These findings suggest that inflammation contributes to brain injury in lacunar stroke. PMID- 11935049 TI - The stroke patient who woke up: clinical and radiological features, including diffusion and perfusion MRI. AB - BACKGROUND AND PURPOSE: Time of stroke onset is uncertain for patients who wake from sleep with stroke. Functional imaging techniques may allow estimation of benefit and risk of acute stroke therapy. We compared the clinical and multimodal MRI findings of patients with uncertain stroke onset with those with known onset time. METHODS: Patients imaged within 24 hours of ischemic stroke onset between January 1997 and June 2000 were identified from a prospective stroke registry. Clinical and imaging data from patients with known stroke onset (group I) were compared with those who woke with stroke (group II). RESULTS: A total of 364 patients were identified, of whom 100 (27%) woke from sleep with stroke. Group I and group II did not differ in age, gender, National Institutes of Health Stroke Scale, or TOAST (Trial of Org 10172 in Acute Stroke Treatment) diagnoses. Time from stroke onset was shorter in group I (mean 6.0 versus 13.3 hours, P<0.001); time from detection did not differ between groups (6.0 versus 5.9 hours). Within 3 hours, diffusion-weighted imaging (DWI) and perfusion-weighted imaging (PWI) lesion volumes were similar in both groups; DWI-PWI mismatch was present in 82% of group I and 73% of group II patients. Mean apparent diffusion coefficient of water (ADC) of group I patients was negatively associated with DWI volume (beta= 0.324, P=0.004) and time from stroke onset (beta=-0.238, P=0.031) in multivariate analysis. The mean ADC of group II patients was lower than that of group I patients within 3 hours of stroke detection (mean 556 versus 665 microm2/s, P<0.01), but individual group II patients had ADC values as high as 742 microm2/s, in addition to a DWI-PWI mismatch pattern. CONCLUSIONS: Onset time is uncertain in over one quarter of acute ischemic stroke patients. Clinical features of these patients do not differ significantly from those with known onset time. Some patients who wake with stroke seem to have favorable imaging characteristics for acute stroke therapy. Further study is needed to determine whether criteria for therapy based on imaging parameters can safely be applied to these patients. PMID- 11935050 TI - Blood use in cerebrovascular neurosurgery. AB - BACKGROUND AND PURPOSE: This study reviews the perioperative use of red blood cell transfusion in cerebrovascular neurosurgery. The current algorithm for preoperative ordering of red cells is historical and dated. More blood is ordered than is actually transfused, and considerable variability exists between different institutions. We determine the use of blood transfusion in cerebrovascular surgery to develop a rational blood ordering practice. METHODS: Records of 301 patients undergoing cerebrovascular neurosurgery at the University of Virginia were reviewed to quantitatively evaluate red blood cell transfusion practices. The amount and reason for transfusion were noted in each case. RESULTS: In 126 patients undergoing carotid endarterectomy, there were no preoperative or intraoperative transfusions and 5 postoperative transfusions (4.0%). In 71 ruptured aneurysm patients, there were 2 preoperative blood transfusions (2.8%), 4 intraoperative transfusions (5.6%), and 15 postoperative transfusions (21.1%). Forty-seven patients underwent surgery for unruptured aneurysms, with no preoperative transfusions, 2 intraoperative transfusions (4.3%), and 8 postoperative blood transfusions (17.0%). Of the 54 patients undergoing surgery for arteriovenous malformations, 5 patients (9.3%) were transfused preoperatively, 4 were transfused intraoperatively (7.4%), and 22 were transfused postoperatively (40.7%). None of the 3 patients undergoing surgery for concomitant arteriovenous malformations and aneurysms received intraoperative blood transfusions, but 1 received blood both preoperatively and postoperatively, and another received a transfusion postoperatively only. The overall ratio of perioperative cross-match to transfusion in this series is 41.4. CONCLUSIONS: In vascular neurosurgery at our institution, blood has routinely been ordered excessively. We recommend an ABO-Rh type and antibody screen for aneurysm and arteriovenous malformation surgery and no screen for carotid endarterectomy to efficiently utilize transfusion therapy in cerebrovascular surgery. PMID- 11935051 TI - Effects of induced hypertension on intracranial pressure and flow velocities of the middle cerebral arteries in patients with large hemispheric stroke. AB - BACKGROUND AND PURPOSE: Our aim was to prospectively evaluate the effects of induced arterial hypertension in patients with large ischemic stroke. METHODS: A total of 47 monitoring sessions in 19 patients with acute, complete, or subtotal middle cerebral artery (MCA) territory stroke were performed. Intracranial pressure (ICP) was monitored using a parenchymal catheter. Mean arterial blood pressure (MAP), ICP, and peak mean flow velocity of the middle cerebral arteries (V(m)MCA) were continuously recorded. Patients with acute ICP crises were excluded. After obtaining baseline values, MAP was raised by an infusion of norepinephrine to reach an MAP increase of at least 10 mm Hg. After MAP had reached a peak plateau level, the norepinephrine infusion was stopped. RESULTS: Baseline MAP was 83.6+/-1.6 mm Hg and rose to 108.9+/-2.0 mm Hg after infusion of norepinephrine. ICP slightly increased from 11.6+/-0.9 mm Hg to 11.8+/-0.9 mm Hg (P<0.05). Cerebral perfusion pressure rose from baseline 72.2+/-2 mm Hg to 97+/-1 mm Hg (P<0.0001). V(m)MCA was already higher on the affected side during baseline measurements. At maximum MAP levels, V(m)MCA rose by 25.5+/-5.5 cm/s on the affected side and by 8.6+/-1.6 cm/s on the contralateral side. CONCLUSIONS: In patients with large hemispheric stroke without an acute ICP crisis, induced hypertension enhances cerebral perfusion pressure and augments the V(m)MCA(s), more so on the affected side. The ICP slightly increases; however, this is probably not clinically significant. PMID- 11935052 TI - High plasma brain natriuretic polypeptide level as a marker of risk for thromboembolism in patients with nonvalvular atrial fibrillation. AB - BACKGROUND AND PURPOSE: Assessment of left atrial appendage (LAA) function with transesophageal echocardiography is useful for detecting patients at high risk for thromboembolism as a result of atrial fibrillation (AF). A recent study reported that the atrium is the main source of brain natriuretic polypeptide (BNP) in AF patients without overt heart failure. The purpose of this study was to assess a possible relationship between LAA function and plasma BNP levels in nonvalvular AF. METHODS: Thirty-four consecutive patients with chronic nonvalvular AF (age, 69+/-9 years) underwent transesophageal echocardiography and plasma BNP measurement. Thirteen patients with a history of thromboembolism or echocardiographic evidence of thrombus (E + group) were compared with 21 AF patients without complications (E- group). RESULTS: The E+ group patients demonstrated greater impairment of LAA velocity and higher plasma BNP levels than the E- group patients (LAA velocity: 12+/-6 versus 31+/-17 cm/s, P<0.05; plasma BNP: 126+/-53 versus 86+/-45 ng/L, P<0.05). Overall analysis of the continuous variables with multiple logistic regression analysis revealed that BNP was a significant predictor of thromboembolism. There was a weak but significant negative correlation between plasma BNP levels and LAA flow velocity (r=0.38, P<0.05). No intergroup difference in plasma atrial natriuretic polypeptide levels was found. CONCLUSIONS: The present data suggest the usefulness of measuring plasma BNP levels, which may reflect augmented atrial secretion of BNP from the impaired atrial myocardium, in detecting patients at high risk for thromboembolic complications in nonvalvular AF. PMID- 11935053 TI - Efficacy and safety of nicardipine prolonged-release implants for preventing vasospasm in humans. AB - BACKGROUND AND PURPOSE: Despite extensive investigative efforts, there are few treatments that can prevent vasospasm after subarachnoid hemorrhage. This study was conducted to examine the efficacy and safety of nicardipine prolonged-release implants (NPRI) for humans, which have already been proven in dogs. METHODS: Twenty consecutive subarachnoid hemorrhage patients with thick subarachnoid clot were treated with NPRI (a pellet of diameter 2 mm, length 10 mm, containing 4 mg of nicardipine) during surgery after clipping of their aneurysm. The number and location of pellets depended on the amount and site of subarachnoid clot on preoperative CT and on craniotomy. RESULTS: Two to 10 pellets were implanted in the cistern of the internal carotid, middle cerebral, and/or anterior cerebral artery, where thick clots existed and therefore vasospasm related to delayed ischemic neurological deficits was highly likely. Delayed ischemic neurological deficits and cerebral infarctions were seen in 1 patient. Angiography performed on days 7 to 12 revealed no vasospasm in any arteries near which NPRI were placed. No complications were experienced. CONCLUSIONS: Vasospasm was completely prevented for the arteries in thick clot cisterns, when NPRI were placed adjacent to the arteries during surgery. This drug-delivery system offers a promising approach for preventing vasospasm. PMID- 11935055 TI - Analysis and comparison of the psychometric properties of three balance measures for stroke patients. AB - BACKGROUND AND PURPOSE: This study compared the psychometric properties of 3 clinical balance measures, the Berg Balance Scale (BBS), the Balance subscale of the Fugl-Meyer test (FM-B), and the Postural Assessment Scale for Stroke Patients (PASS), in stroke patients with a broad range of neurological and functional impairment from the acute stage up to 180 days after onset. METHODS: One hundred twenty-three stroke patients were followed up prospectively with the 3 balance measures 14, 30, 90, and 180 days after stroke onset (DAS). Reliability (interrater reliability and internal consistency) and validity (concurrent validity, convergent validity, and predictive validity) of each measure were examined. A comparison of the responsiveness of each of the 3 measures was made on the basis of the entire group of patients and 3 separate groups classified by degree of neurological severity. RESULTS: The FM-B and BBS showed a significant floor or ceiling effect at some DAS points, whereas the PASS did not show these effects. The BBS, FM-B, and PASS all had good reliability and validity for patients at different recovery stages after stroke. The results of effect size demonstrated fair to good responsiveness of all 3 measures within the first 90 DAS but, as expected, only a low level of responsiveness at 90 to 180 DAS. The PASS was more responsive to changes in severe stroke patients at the earliest period after stroke onset, 14 to 30 DAS. CONCLUSIONS: All 3 measures tested showed very acceptable levels of reliability, validity, and responsiveness for both clinicians and researchers. The PASS showed slightly better psychometric characteristics than the other 2 measures. PMID- 11935054 TI - Well-being after stroke in Canadian seniors: findings from the Canadian Study of Health and Aging. AB - BACKGROUND AND PURPOSE: After a stroke many people continue to live with their residual impairments and disabilities in the community, which can pose a significant problem for survivors' well-being. The purpose of this research was to investigate patterns of well-being in community-dwelling stroke survivors to identify those factors that restrict and enhance well-being. METHODS: A secondary analysis was conducted on data from the second wave of the Canadian Study of Health and Aging (CSHA-2). A national sample of 5395 community-dwelling Canadian seniors (aged > or =65 years) was interviewed for CSHA-2, including 339 stroke survivors (6.3%). Information was collected on health, social and demographic characteristics, and well-being. Comparisons were made between the health and functional status of stroke survivors and community-dwelling seniors who have not experienced a stroke. Multiple regression was used to examine the factors associated with well-being in stroke survivors. RESULTS: Compared with community dwelling seniors who have not experienced a stroke, stroke survivors report a lower sense of well-being. Stroke survivors are also more likely to be restricted in their physical and cognitive function, to report worse mental health, and to be living with a greater number of comorbid health conditions. Mental health and physical and cognitive disabilities are associated with a reduced sense of well being in stroke survivors, but social supports and educational resources moderate the impact of functional status on well-being. CONCLUSIONS: Community-dwelling seniors who have had a stroke experience a reduced sense of well-being. However, social resources can help to alleviate the subjective burden of this common neurological condition. PMID- 11935056 TI - Informal care for stroke survivors: results from the North East Melbourne Stroke Incidence Study (NEMESIS). AB - BACKGROUND AND PURPOSE: Informal caregivers play an important role in the lives of stroke patients, but the cost of providing this care has not been estimated. The purpose of this study was to determine the nature and amount of informal care provided to stroke patients and to estimate the economic cost of that care. METHODS: The primary caregivers of stroke patients registered in the North East Melbourne Stroke Incidence Study (NEMESIS) were interviewed at 3, 6, and 12 months after stroke, and the nature and amount of informal care provided were documented. The opportunity and replacement costs of informal care for all first ever-in-a-lifetime strokes (excluding subarachnoid hemorrhages) that occurred in 1997 in Australia were estimated. RESULTS: Among 3-month stroke survivors, 74% required assistance with activities of daily living and received informal care from family or friends. Two thirds of primary caregivers were women, and most primary caregivers (>90%) provided care during family or leisure time. Total first-year caregiver time costs for all first-ever-in-a-lifetime strokes were estimated to be A$21.7 million (opportunity cost approach) or A$42.5 million (replacement cost approach), and the present values of lifetime caregiver time costs were estimated to be A$171.4 million (opportunity cost approach) or A$331.8 million (replacement cost approach). CONCLUSIONS: Informal care for stroke survivors represents a significant hidden cost to Australian society. Because our community is rapidly aging, this informal care burden may increase significantly in the future. PMID- 11935057 TI - Long-term disability after first-ever stroke and related prognostic factors in the Perth Community Stroke Study, 1989-1990. AB - BACKGROUND AND PURPOSE: Few reliable estimates of the long-term functional outcome after stroke are available. This population-based study aimed to describe disability, dependency, and related independent prognostic factors at 5 years after a first-ever stroke in patients in Perth, Western Australia. METHODS: All individuals with a suspected acute stroke who were resident in a geographically defined region (population, 138 708) of Perth, Western Australia, were registered prospectively and assessed according to standardized diagnostic criteria over a period of 18 months in 1989 to 1990. Patients were followed up prospectively at 4 and 12 months and 5 years after the index event. RESULTS: There were 370 cases of first-ever stroke, and 277 patients survived to 30 days. Of these early survivors, 152 (55%) were alive at 5 years, and among those who were neither institutionalized (n=146) nor disabled (n=129) at the time of their stroke, 21 (14%) were institutionalized in a nursing home, and 47 (36%) were disabled. The most important predictors of death or disability at 5 years were increasing age, baseline disability defined by a Barthel Index score of <20/20 (odds ratio [OR], 6.3; 95% confidence interval [CI], 2.7 to 14), moderate hemiparesis (OR, 2.7; 95% CI, 1.1 to 6.2), severe hemiparesis (OR, 4.5; 95% CI, 1.1 to 19), and recurrent stroke (OR, 9.4; 95% CI, 3.0 to 30). A low level of activity before the stroke was a significant predictor of institutionalization, and subsequent recurrent stroke was a consistent, independent predictor of institutionalization, disability, and death or institutionalization, increasing the odds of each of these 3 adverse outcomes by 5- to 15-fold. CONCLUSIONS: Among 30-day survivors of first-ever stroke, about half survive 5 years; of survivors, one third remain disabled, and 1 in 7 are in permanent institutional care. The major modifiable predictors of poor long-term outcome are a low level of activity before the stroke and subsequent recurrent stroke. Efforts to increase physical activity among the elderly and to prevent recurrent stroke in survivors of a first stroke are likely to reduce the long-term burden of cerebrovascular disease. PMID- 11935058 TI - Predicting outcome after acute and subacute stroke: development and validation of new prognostic models. AB - BACKGROUND AND PURPOSE: Statistical models to predict the outcome of patients with acute and subacute stroke could have several uses, but no adequate models exist. We therefore developed and validated new models. METHODS: Regression models to predict survival to 30 days after stroke and survival in a nondisabled state at 6 months were produced with the use of established guidelines on 530 patients from a stroke incidence study. Three models were produced for each outcome with progressively more detailed sets of predictor variables collected within 30 days of stroke onset. The models were externally validated and compared on 2 independent cohorts of stroke patients (538 and 1330 patients) by calculating the area under receiver operating characteristic curves (AUC) and by plotting calibration graphs. RESULTS: Models that included only 6 simple variables (age, living alone, independence in activities of daily living before the stroke, the verbal component of the Glasgow Coma Scale, arm power, ability to walk) generally performed as well as more complex models in both validation cohorts (AUC 0.84 to 0.88). They had good calibration but were overoptimistic in patients with the highest predicted probabilities of being independent. There were no differences in AUCs between patients seen within 48 hours of stroke onset and those seen later; between ischemic and hemorrhagic strokes; and between those with and without a previous stroke. CONCLUSIONS: The simple models performed well enough to be used for epidemiological purposes such as stratification in trials or correction for case mix. However, clinicians should be cautious about using these models, especially in hyperacute stroke, to influence individual patient management until they have been further evaluated. Further research is required to test whether additional information from brain imaging improves predictive accuracy. PMID- 11935059 TI - Prognostic significance of uric acid serum concentration in patients with acute ischemic stroke. AB - BACKGROUND AND PURPOSE: We sought to assess in 881 consecutive patients with acute ischemic stroke the clinical relevance in regard to functional outcome of the natural antioxidant uric acid measured at hospital admission. METHODS: Patients had serum uric acid (mg/dL) measured by standard procedures 18.2+/-15.5 hours from clinical onset. At hospital discharge (11.0+/-6.0 days), neurological impairment was classified as moderate/severe (Mathew score < or =75; n=304) or mild/absent (Mathew score >75; n=577). Demographics, atherosclerotic risk factors, history of organ disease, baseline neurological score, stroke subtype, infarction size, renal function, aspirin use before stroke, stroke therapy, diuretic use, and laboratory markers, including erythrocyte sedimentation rate, were analyzed in both outcome groups with the use of backward logistic regression. RESULTS: Increased uric acid values were found in men, hypertensives, alcohol drinkers, and patients with coronary, pulmonary, or renal diseases. Diabetic patients had lower uric acid levels on admission. Uric acid was directly associated with hematocrit (P=0.001), sodium (P=0.001), creatinine (P=0.001), and triglycerides (P=0.001) and inversely related with nonfasting glucose (P=0.001) levels. Neurological impairment on admission (P=0.001) and final infarction size on CT/MRI (P=0.01) were also inversely associated with uric acid. A logistic regression adjusted for confounders confirmed the following independent (odds ratio, 95% CI) good outcome predictors: age (0.97, 0.96 to 0.99), Mathew score on admission (1.14, 1.12 to 1.17), erythrocyte sedimentation rate (0.98, 0.97 to 0.99), infarction volume (0.98, 0.98 to 0.99), and uric acid (1.12, 1.00 to 1.25). CONCLUSIONS: In patients with acute ischemic stroke, there is a 12% increase in the odds of good clinical outcome for each milligram per deciliter increase of serum uric acid. This finding reinforces the relevance of oxidative damage in ischemic stroke. PMID- 11935060 TI - Dysplastic vessels after surgery for brain arteriovenous malformations. AB - BACKGROUND AND PURPOSE: The cause and clinical significance of residual dysplastic vessels after surgery for brain arteriovenous malformations (AVM) are unclear. We studied predictors and frequency of residual dysplastic vessels on cerebral angiography after AVM surgery. METHODS: The 240 prospectively enrolled surgical patients from the New York AVM Databank underwent 269 AVM-related surgical procedures. Reported postoperative brain angiographic findings were classified post hoc as showing (1) persistent dysplastic vessels, (2) a residual AVM, (3) focal hyperemia in the surgical bed, (4) other changes, or (5) a normal angiogram. Univariate and multivariate models were applied to test for an association between residual dysplastic vessels and patient age, sex, preoperative AVM size, anatomic AVM location, number of embolization procedures before surgery, and the time interval between AVM surgery and the postoperative angiogram. RESULTS: Of the 224 documented postoperative angiograms, 78 (35%) showed dysplastic vessels, 24 (11%) had evidence for a residual AVM, 16 (7%) showed focal hyperemia, 6 (2%) revealed other findings, and 100 (45%) were normal. The number of cases showing angiographic evidence for dysplastic vessels was significantly associated with increasing size of the AVM (in millimeter increments; P=0.0001); the mean diameter of AVMs in patients showing dysplastic vessels after surgery was significantly larger (41 mm, SD +/-14) than in those without residual dysplastic vessels (27 mm, SD +/-13; P<0.001). Symptomatic postoperative intracerebral hemorrhage occurred in 4 patients (1%), in 2 of whom dysplastic vessels were seen on the postoperative angiogram. CONCLUSIONS: The findings suggest that persistent dysplastic vessels may be found in approximately one third of angiograms after AVM surgery. Preoperative AVM size was found to be an independent predictor for the occurrence of dysplastic vessels on the postoperative angiogram. PMID- 11935061 TI - Risk of stroke, transient ischemic attack, and vessel occlusion before endarterectomy in patients with symptomatic severe carotid stenosis. AB - BACKGROUND AND PURPOSE: We aimed to identify and determine the clinical relevance of parameters predictive of stroke recurrence and vessel occlusion before carotid endarterectomy. METHODS: One hundred forty-three consecutive patients (105 men; mean age, 66.1+/-8 years) with symptomatic severe carotid artery stenosis were prospectively followed up until carotid endarterectomy. Patients had suffered an ischemic vascular event in the ipsilateral anterior circulation 9.6 days (median; range, 0 to 92 days) before presentation and assessment of stenosis. Admission examination included medical history, neurological status, extracranial and transcranial Doppler/duplex sonography, CT/MRI, ECG, and routine laboratory examination. All patients were reevaluated in the same way the day before surgery (without CT/MRI) and at recurrence of an ischemic event (including CT/MRI). RESULTS: The end point of follow-up after 19.0 days (median; range, 0 to 118) was carotid endarterectomy in 120 patients, ipsilateral recurrent ischemia in 15 patients (7 transient events and 8 disabling strokes, with carotid occlusion in 4), and (asymptomatic) carotid occlusion in 8 patients. An exhausted cerebrovascular reactivity as determined by a Doppler CO2 test in the middle cerebral artery ipsilateral to the stenosis was the only independent predictive parameter for disabling stroke (odds ratio [OR], 9.7; 95% confidence interval [CI], 2.1 to 44.1; P=0.003). Stroke rate in patients with exhausted reactivity was 27% per month compared with 5.2% in those with normal reactivity. Progression of stenosis toward occlusion was observed in 12 patients and correlated with decreased poststenotic peak systolic velocity (OR, 0.75; 95% CI, 0.62 to 0.90; P=0.002), poststenotic arterial narrowing (OR, 22.7; 95% CI, 3.6 to 141.6; P=0.001), and very severe stenosis (OR, 13.6; 95% CI, 2.2 to 83.7; P=0.005). In patients without hemodynamic compromise, occlusion was not associated with increased stroke risk. CONCLUSIONS: Patients with recently symptomatic high-grade carotid artery stenosis and ipsilateral hemodynamic compromise are at high risk for early disabling stroke. Assessment of the hemodynamic status is recommended after diagnosis of severe carotid stenosis in symptomatic patients to further investigate and evaluate whether these patients may benefit from early endarterectomy. PMID- 11935062 TI - Carotid stenting and endarterectomy: a clinical and cost comparison of revascularization strategies. AB - BACKGROUND AND PURPOSE: Investigational carotid stenting for extracranial carotid stenosis has demonstrated procedural results approaching those reported with endarterectomy, but with limited cost and long-term data. This study compared the in-hospital outcomes and costs of these 2 revascularization strategies at a single institution and the long-term effectiveness of carotid stenting. METHODS: Data for 136 endarterectomies and 136 carotid stent procedures at a tertiary-care community hospital were obtained. The primary clinical outcome measures were in hospital major ipsilateral stroke and death. In-hospital direct variable costs and length of stay were the primary economic measures. Information on restenosis and late ipsilateral stroke for the stent group was available at 2-year follow up. RESULTS: These nonrandomized groups were similar, but the endarterectomy group had more symptomatic patients (42% versus 31%; P=0.0004), and the stent group had more NASCET-excluded patients (68% versus 35%; P<0.0001). In-hospital major ipsilateral stroke and death occurred more frequently in the surgical group, but the difference was not significant (2.9% versus 0%; P=0.1). Minor ipsilateral strokes were similar (2.2% versus 2.9%; P=NS). Cost ($5409 versus $3417; P<0.0001) and length of stay (3.0 versus 1.4 days; P<0.0001) were significantly greater for the surgical group. In the stent group, 6-month angiographic restenosis was 3.1%, and 2-year ipsilateral major stroke rate was 0%. CONCLUSIONS: In-hospital outcomes with carotid stenting were similar to those with endarterectomy but were achieved in patients with significantly more comorbidities. Cost and resource utilization with stenting were substantially less than those with endarterectomy. At 2 years, carotid stenting appeared not only durable but also effective in stroke prevention. PMID- 11935063 TI - Expression and function of recombinant S1179D endothelial nitric oxide synthase in canine cerebral arteries. AB - BACKGROUND AND PURPOSE: Bovine endothelial nitric oxide synthase (eNOS) is phosphorylated directly by the protein kinase Akt at serine 1179. Mutation of this residue to the negatively charged aspartate (S1179DeNOS) increases nitric oxide (NO) production constitutively in the absence of agonist stimulus. The present study was designed to determine the effect of mutant S1179DeNOS gene expression on vasomotor function of canine cerebral arteries. METHODS: Isolated basilar and middle cerebral arteries were exposed ex vivo (30 minutes at 37 degrees C) to an adenoviral vector (10(10) plaque-forming units per milliliter) encoding the S1179DeNOS gene (AdCMVS1179DeNOS), the wild-type eNOS gene (AdCMVeNOS), or the green fluorescent protein (GFP) reporter gene (AdCMVGFP). Twenty-four hours after transduction, arteries were suspended in an organ chamber for isometric force recording, and levels of cGMP were measured by radioimmunoassay. RESULTS: Transgene protein expression was detected mainly in the vascular adventitia. In AdCMVS1179DeNOS-transduced arteries, basal levels of cGMP were significantly elevated compared with those in control (nontransduced), AdCMVGFP-, or AdCMVeNOS-transduced vessels (n=8; P<0.01). The elevation of cGMP was abolished by a NOS inhibitor, N(G)-nitro-L-arginine methyl ester (L-NAME), or by incubation in the calcium-free medium in the presence of calcium chelators. In AdCMVS1179DeNOS-transduced arteries, contractions to endothelin-1 (10(-10) to 10( 8) mol/L) were significantly reduced compared with those in control and AdCMVGFP transduced arteries (n=7; P<0.05). The vasoconstrictor effect of endothelin-1 was restored in the presence of the NOS inhibitor L-NAME. CONCLUSIONS: Our results suggest that in cerebral arteries, expression of recombinant S1179DeNOS increases basal production of NO and inhibits the vasoconstrictor effect of endothelin-1. This effect may have therapeutic application in prevention and treatment of cerebrovascular diseases. PMID- 11935064 TI - Albumin therapy of transient focal cerebral ischemia: in vivo analysis of dynamic microvascular responses. AB - BACKGROUND AND PURPOSE: To study whether intravascular or hemodynamic factors contribute to the marked neuroprotective effect of albumin therapy in focal cerebral ischemia, 2 complementary methods were applied: laser-scanning confocal microscopy (LSCM) and laser-Doppler perfusion imaging (LDPI). METHODS: In the LSCM study, Sprague-Dawley rats were anesthetized with halothane/nitrous oxide, and a cranial window was placed over the dorsolateral frontoparietal cortex. Rats received 2-hour middle cerebral artery occlusion (MCAO) by an intraluminal suture and were treated with human albumin (1.25 g/kg; n=4) or saline (n=3) after 30 minutes of recirculation. Video images of cortical vessels were continually acquired and were digitized offline to measure diameters and fluorescent erythrocyte velocities. In the LDPI study, cortical perfusion was measured in anesthetized Sprague-Dawley rats that received 2-hour MCAO and were treated with albumin (2.5 g/kg; n=6) or saline (n=5) at 30 minutes after recirculation. RESULTS: In the LSCM study, MCAO was associated with arteriolar dilation and slowing of capillary and venular erythrocyte perfusion. During the first 15 to 30 minutes of postischemic recirculation, prominent foci of vascular stagnation developed within cortical venules, associated with thrombuslike foci and adherent corpuscular structures consistent in size with neutrophils. Saline administration failed to affect these phenomena, while albumin therapy was followed by significant increases in arteriolar diameter ( approximately 12%; P=0.007) and by a prompt improvement of venular and capillary erythrocyte perfusion and a partial disappearance of adherent thrombotic material. Albumin therapy increased erythrocyte flow velocity in both capillaries (288+/-73% versus 76+/-18% in the saline group; P=0.023) and venules (2.7-fold [P=0.001] versus 1.0-fold in the saline group [P=NS]). In the LDPI study, cortical perfusion declined during MCAO and rose initially with recirculation (to approximately 135% of baseline) in both groups. Mean cortical perfusion improved slightly (approximately 14%; P=NS) in albumin-treated animals. CONCLUSIONS: These results reveal a beneficial effect of albumin therapy in reversing stagnation, thrombosis, and corpuscular adherence within cortical venules in the reperfusion phase after focal ischemia and support its utility in the treatment of acute ischemic stroke. PMID- 11935065 TI - NAP, a femtomolar-acting peptide, protects the brain against ischemic injury by reducing apoptotic death. AB - BACKGROUND AND PURPOSE: We sought to determine the cerebroprotective potential of NAP, a synthetic octapeptide related to vasoactive intestinal peptide. Activity dependent neuroprotective protein mediates some of the protective effects of vasoactive intestinal peptide. The neuroprotective NAP sequence is derived from activity-dependent neuroprotective protein. METHODS: Spontaneously hypertensive rats underwent permanent middle cerebral artery occlusion by craniotomy and electrocoagulation. After dose-response and time-course experiments, the animals were injected with NAP (3 microg/kg) or vehicle intravenously 1 hour after stroke onset. Another group of rats was injected with the D-amino acid isomer of NAP (D NAP) and served as a negative control. Rats were examined for motor and behavioral deficits 24 hours to 30 days later, and infarct volumes were determined. The effect of NAP administration on apoptotic death was determined by terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling (TUNEL) and caspase-3 stainings. RESULTS: NAP significantly reduced motor disability and infarct volumes compared with vehicle or D-NAP when tested at 24 hours after stroke onset (9.67+/-1.4% versus 17.04+/-1.18% and 19.19+/-1.9% of hemispheric volume, respectively; P<0.05). NAP given 4 but not 6 hours after permanent middle cerebral artery occlusion still conferred significant neuroprotection (infarct volume 10.9+/-3.9% of hemispheric volume; P<0.05 versus vehicle). Long-term studies demonstrated that infarct volumes and disability scores remained significantly lower after 30 days in NAP-treated animals. NAP significantly reduced the number of apoptotic cells. CONCLUSIONS: Our results indicate that the durable cerebroprotection by NAP involves antiapoptotic mechanisms. PMID- 11935066 TI - Ischemia and ischemic tolerance induction differentially regulate protein expression of GluR1, GluR2, and AMPA receptor binding protein in the gerbil hippocampus: GluR2 (GluR-B) reduction does not predict neuronal death. AB - BACKGROUND AND PURPOSE: Postischemic delayed neuronal death (DND) of hippocampal CA1 neurons has been suggested to occur as a result of formation of calcium permeable alpha-amino-3-hydroxy-5-methyl-4-isoxazole propionic acid (AMPA) receptors lacking the GluR2 subunit (GluR2 hypothesis). DND can be prevented by a short tolerance-inducing ischemic period. The present study was designed to assess whether postischemic protein levels of GluR2 predict neuronal death. Additionally, the role of AMPA receptor binding protein (ABP) was investigated with respect to neuronal death or survival. METHODS: Postischemic protein expression of GluR1, GluR2, and ABP was analyzed in 3 experimental paradigms of transient global ischemia with the use of subunit-specific antisera and semiquantitative densitometric evaluation. Gerbils were subjected (1) to a 5 minute ischemic period resulting in DND of CA1 neurons; (2) to a 2.5-minute period of ischemia mediating tolerance induction; and (3) to 5 minutes of ischemia in the ischemia-tolerant state (2.5+5 minutes of ischemia 4 days apart). RESULTS: The major finding was that GluR2 protein levels were significantly downregulated in neuronal subpopulations destined to survive, ie, in CA1 principal neurons after ischemic tolerance induction and in the ischemia-tolerant state, as well as in CA3 neurons after a 5-minute period of ischemia. ABP expression remained unaffected. CONCLUSIONS: Our results modify the GluR2 hypothesis in that postischemic GluR2 reduction also occurs in hippocampal CA1 and CA3 principal neurons without subsequent neuronal death. ABP is obviously not involved in mechanisms of DND or ischemic tolerance induction. PMID- 11935067 TI - Poly(ADP-ribose) polymerase impairs early and long-term experimental stroke recovery. AB - BACKGROUND AND PURPOSE: Poly(ADP-ribose) polymerase (PARP-1; Enzyme Commission 2.4.30) is a nuclear DNA repair enzyme that mediates early neuronal ischemic injury. Using novel 3-dimensional, fast spin-echo-based diffusion-weighted imaging, we compared acute (21 hours) and long-term (3 days) ischemic volume after middle cerebral artery (MCA) occlusion in PARP-1-null mutants (PARP-/-) versus genetically matched wild-type mice (WT mice). PARP-/- mice were also treated with viral transfection of wild-type PARP-1 to determine whether protection from MCA occlusion is lost with restoration of the gene product. METHODS: Halothane-anesthetized mice were treated with reversible MCA occlusion via intraluminal suture technique. Ischemic volumes were delineated by diffusion weighted imaging with high spatial and temporal resolution during MCA occlusion and reperfusion. Recombinant Sindbis virus carrying beta-galactosidase (lacZ) or PARP-1 was injected into ipsilateral striatum, then animals underwent MCA occlusion 3 days later. Infarction volume was measured at 22 hours of reperfusion (2,3,5-triphenyltetrazolium chloride histology). RESULTS: Reduction in regional water apparent diffusion coefficient (ADC) during occlusion or secondary ADC decline during reperfusion was not different between groups. Ischemic volume was smaller early in occlusion in PARP-/- versus WT mice and remained less at 21 hours of reperfusion. Ischemic volume then increased from 1 to 2 days in all mice, then stabilized without further change. Ischemic damage was smaller in PARP /- than in WT mice at 3 days. Transfection of PARP-1 into PARP-/- mice increased stroke damage relative to lacZ-injected PARP-/- and increased damage to that of the WT mice. Intraischemic laser-Doppler flowmetry and physiological variables were not different among groups. CONCLUSIONS: PARP-1 deficiency provides both early and prolonged protection from experimental focal stroke. The mechanism is not linked to preservation of ADC and mitigation of secondary energy depletion during early reperfusion. PMID- 11935068 TI - Retinal and optic nerve degeneration after chronic carotid ligation: time course and role of light exposure. AB - BACKGROUND AND PURPOSE: Carotid artery disease can cause chronic retinal ischemia, resulting in transient or permanent blindness, pupillary reflex dysfunction, and retinal degeneration. This experiment investigated the effects of chronic retinal ischemia in an animal model involving permanent carotid occlusion. The time course of retinal pathology and the role of light in this pathology were examined. METHODS: Sprague-Dawley rats underwent permanent bilateral occlusion of the common carotid arteries or sham surgery. Half of the animals were postsurgically housed in darkness, and half were housed in a 12-hour light/dark cycle. Animals were killed at 3, 15, and 90 days after surgery. Stereological techniques were used to count the cells of the retinal ganglion cell layer. Thy-1 immunoreactivity was assessed to specifically quantify loss of retinal ganglion cells. The thicknesses of the remaining retinal sublayers were measured. Optic nerve degeneration was quantified with the Gallyas silver staining technique. RESULTS: Permanent bilateral occlusion of the common carotid arteries resulted in loss of the pupillary reflex to light in 58% of rats. Eyes that lost the reflex showed (1) optic nerve degeneration at 3, 15, and 90 days after surgery; (2) a reduction of retinal ganglion cell layer neurons and Thy-1 immunoreactivity by 15 and 90 days; and (3) a severe loss of photoreceptors by 90 days when postsurgically housed in the light condition only. CONCLUSIONS: Ischemic damage to the optic nerve caused loss of pupillary reflex and death of retinal ganglion cells in a subset of rats. Subsequently, light toxicity induced death of the photoreceptors. PMID- 11935069 TI - Temporal profile of enhanced vulnerability of the postthrombotic brain to secondary embolic events. AB - BACKGROUND AND PURPOSE: Patients with vascular or cardiac disease may experience recurrent thrombosis and embolization to the cerebral vasculature. Transient distal platelet accumulation after common carotid artery thrombosis (CCAT) leads to hemodynamic, metabolic, and molecular events that may influence the response of the postthromboembolic brain to secondary emboli. We investigated the effect of repeated embolic episodes on histopathological outcome at various time intervals using a clinically relevant model of embolic stroke. METHODS: Six groups of rats underwent either photochemically induced CCAT followed by sham surgery or 2 episodes of CCAT separated by 10 minutes or 1, 3, 5, or 7 days. Outcome measures included routine histopathological analysis and determination of the number of infarct loci and their total volume. RESULTS: Rats that underwent a second CCAT at 1, 3, or 5 days after the first insult had 20 to 30 times larger infarct volumes than rats in the single-CCAT group (P<0.05). In addition, rats in the 10-minute and 1-, 3-, and 5-day groups had 2 to 3 times as many infarcts as those in the single-CCAT group (P<0.05). Infarcts produced by double insults commonly extended through the neuraxis and were necrotic, edematous, and sometimes hemorrhagic. CONCLUSIONS: A prior thromboembolic event puts the brain at risk for severe infarction after a second embolic event. These findings cannot be explained solely by a greater number of infarcts. Elucidating pathomechanisms responsible for the vulnerability of the postthromboembolic brain may provide targets for new treatment strategies to prevent the severe consequences of embolic stroke. PMID- 11935070 TI - Cellular responses of bioabsorbable polymeric material and Guglielmi detachable coil in experimental aneurysms. AB - BACKGROUND AND PURPOSE: Acceleration of healing mechanisms is a promising approach to improve current limitations of endovascular aneurysm therapy with the use of platinum coils. We evaluated a new endovascular therapeutic, bioabsorbable polymeric material (BPM), which may promote cellular reaction in the aneurysms. METHODS: Four different concentrations of lactide/glycolic acid copolymer [poly(D L-lactic-co-glycolic acid)] (PLGA), 85/15, 75/25, 65/35, and 50/50, were used as BPMs. Sixteen experimental aneurysms were created in 8 swine. Eight-millimeter long spiral-shaped BPMs were surgically implanted in the aneurysms without tight packing (n=3 for each BPM). Guglielmi detachable coils (GDCs) were used as control (n=4). The animals were killed 14 days after embolization, and angiographic, histological, and immunohistochemical analyses were performed. RESULTS: Despite loose packing of aneurysms with BPMs, faster BPMs such as 50/50 or 65/35 PLGA demonstrated more mature collagen formation and fibrosis in the sac and neck of the aneurysm. One aneurysm treated with 65/35 PLGA, 1 treated with 75/25 PLGA, and all 3 treated with 85/15 PLGA showed a neck remnant on angiography. There was a linear relationship between collagen levels and polymer degradation properties (r=-0.9513). CONCLUSIONS: This preliminary animal study indicates that acceleration of aneurysm healing with the use of BPM is feasible. This concept can be applied to decrease and perhaps prevent aneurysmal recanalization after endovascular treatment of cerebral aneurysms. PMID- 11935071 TI - Dendritic cells are present in ischemic brain after permanent middle cerebral artery occlusion in the rat. AB - BACKGROUND AND PURPOSE: Cerebral ischemia is associated with inflammation involving accumulation of polymorphonuclear neutrophils. T cells have been suggested to contribute to the secondary progression of ischemic brain injury. Dendritic cells (DC) are potent regulators of immunity by activating and tolerizing T cells. DC have previously been detected in rat meninges and choroid plexus. Hypothesizing that DC are involved in inflammation associated with cerebral ischemia, we investigated DC in the brain of Sprague-Dawley rats after permanent middle cerebral artery occlusion (pMCAO) versus sham operation. METHODS: All experimental rats (n=24) had the right MCA permanently occluded by inserting a nylon monofilament through the right external carotid artery. Immunohistochemistry was used to detect DC (OX62(+)), microglia/macrophages (OX42(+)) that developed into DC, and activated DC expressing major histocompatibility complex class II (OX6(+)) in the brain hemispheres at 1 hour to 6 days after pMCAO or sham operation. RESULTS: Levels of DC were elevated at 1 hour in the ischemic versus sham hemispheres (P<0.001) and ischemic versus nonischemic hemispheres (P<0.001). Activated DC expressing major histocompatibility complex class II (OX62(+)OX6(+)) were still elevated at 6 days after pMCAO in the ischemic versus nonischemic hemispheres (P<0.01). The area of brain lesion correlated with numbers of OX62(+) DC per 100-mm2 brain tissue section (r=0.79; P<0.0001). CONCLUSIONS: Increased levels of DC in the brain after pMCAO and correlation between DC numbers and brain lesion area indicate a role for DC in cerebral ischemia. This observation could constitute a basis for further studies on the role of DC in inflammation related to cerebral ischemia. PMID- 11935073 TI - Autoantibodies against oxidatively modified lipoproteins and progression of carotid restenosis after carotid endarterectomy. AB - BACKGROUND: The purpose of this study was to determine whether LDL oxidation could play an important pathogenic role in early restenosis after carotid endarterectomy. An immunologic and biochemical study was performed on a group of patients who underwent carotid endarterectomy, and the degree of intima-media thickness was measured 6 months after surgery. SUMMARY OF REPORT: Fifty-two consecutive patients were included in the study. The presence of antibodies against oxidized LDL in the serum at the time of surgery was evaluated and compared with echo Doppler flow imaging 6 months after the operation. A statistically significant correlation was found between the arterial wall thickness at the site of surgery and the absolute value of IgG antibodies against oxidized LDL (P<0.012) and IgM immunocomplexes (P<0.043). CONCLUSIONS: The presence of antibodies against oxidized LDL at the time of surgery seems to predict a greater intima-media wall hyperplasia at 6 months after surgery, usually recognized as early restenosis. PMID- 11935072 TI - Inflammation and stroke: the Leiden 85-Plus Study . AB - BACKGROUND: Experimental evidence indicates that interleukin-10 (IL-10) deficiency is associated with the development of cardiovascular and cerebrovascular disease. We analyzed the relation between low IL-10 production levels, history of stroke, and incident fatal stroke. SUMMARY OF REPORT: All 85 year-old inhabitants of Leiden, Netherlands (n=599) were visited at their place of residence (response rate, 87%). Production levels of the anti-inflammatory cytokine IL-10 were assessed in a whole blood assay whereby lipopolysaccharide was used as a stimulus. Plasma concentrations of C-reactive protein (CRP) were also used as a marker of inflammation. A history of stroke was obtained at baseline (prevalence, 10%). The number of fatal strokes was prospectively obtained for a median follow-up of 2.6 years (incidence, 1.82 per 100 person years at risk). Subjects with a history of stroke had significantly lower median IL-10 production levels at baseline than subjects without stroke (558 versus 764 pg/mL; P<0.05). They also had significantly higher median CRP concentrations (6 versus 3 mg/L; P<0.05). The odds ratio for a history of stroke increased to 2.30 (95% CI, 1.12 to 4.72) over strata representing decreasing production levels of IL-10. The relative risk for incident fatal stroke was 2.94 (95% CI, 1.01 to 8.53) when we compared subjects with low or intermediate baseline IL-10 production levels to those with high production levels of IL-10. CONCLUSIONS: Our data support the hypothesis that subjects with low IL-10 production levels have an increased risk of stroke. PMID- 11935074 TI - Neonatal cerebral infarction diagnosed by diffusion-weighted MRI: pseudonormalization occurs early. AB - BACKGROUND: Seizures in the neonatal period may be the single symptom of acute ischemic cerebral infarction. It may be difficult to establish the diagnosis in the acute phase by the use of ultrasound, CT, and conventional MRI because of the high water content of the immature brain. Diffusion-weighted (DW) MRI is a very sensitive and fast imaging modality to visualize acute ischemic stroke in infants even before conventional MR images become abnormal. Signal abnormality in DW MRI, however, seems to follow a different time course than in older patients. CASE DESCRIPTION: DW MRI became falsely negative 1 week after stroke (pseudonormalization) in 2 newborn patients during persistence of signal abnormalities on turbo spin-echo images, whereas the so-called pseudonormalization in adults normally occurs within 10 to 14 days. CONCLUSIONS: T2-weighted sequences should supplement DW images to reliably detect subacute ischemic infarctions in the neonatal period. PMID- 11935075 TI - Quantification of perfusion using bolus tracking magnetic resonance imaging in stroke: assumptions, limitations, and potential implications for clinical use. AB - BACKGROUND: MR techniques have been very powerful in providing indicators of tissue perfusion, particularly in studies of cerebral ischemia. There is considerable interest in performing absolute perfusion measurements, with the aim of improving the characterization of tissue "at risk" of stroke. However, some important caveats relating to absolute measurements need to be taken into account. The purpose of this article is to discuss some of the issues involved and the potential implications for absolute cerebral blood flow measurements in clinical use. SUMMARY OF COMMENT: In bolus tracking MRI, deconvolution of the concentration-time course can in theory provide accurate quantification. However, there are several important assumptions in the tracer kinetic model used, some of which may be invalid in cerebral ischemia. These can introduce significant errors in perfusion quantification. CONCLUSIONS: Although we believe that bolus tracking MRI is a powerful technique for the evaluation of perfusion in cerebral ischemia, interpretation of perfusion maps requires caution; this is particularly true when absolute quantification is attempted. Work is currently under way in a number of centers to address these problems, and with appropriate modeling they may be overcome in the future. In the interim, we believe that it is necessary for users of bolus tracking perfusion data to be aware of the current technical limitations if they are to avoid misinterpretation or overinterpretation of their findings. PMID- 11935076 TI - Alzheimer disease as a vascular disorder: nosological evidence. AB - BACKGROUND: The main stumbling block in the clinical management and in the search for a cure of Alzheimer disease (AD) is that the cause of this disorder has remained uncertain until now. SUMMARY OF REVIEW: Evidence that sporadic (nongenetic) AD is primarily a vascular rather than a neurodegenerative disorder is reviewed. This conclusion is based on the following evidence: (1) epidemiological studies showing that practically all risk factors for AD reported thus far have a vascular component that reduces cerebral perfusion; (2) risk factor association between AD and vascular dementia (VaD); (3) improvement of cerebral perfusion obtained from most pharmacotherapy used to reduce the symptoms or progression of AD; (4) detection of regional cerebral hypoperfusion with the use of neuroimaging techniques to preclinically identify AD candidates; (5) presence of regional brain microvascular abnormalities before cognitive and neurodegenerative changes; (6) common overlap of clinical AD and VaD cognitive symptoms; (7) similarity of cerebrovascular lesions present in most AD and VaD patients; (8) presence of cerebral hypoperfusion preceding hypometabolism, cognitive decline, and neurodegeneration in AD; and (9) confirmation of the heterogeneous and multifactorial nature of AD, likely resulting from the diverse presence of vascular risk factors or indicators of vascular disease. CONCLUSIONS: Since the value of scientific evidence generally revolves around probability and chance, it is concluded that the data presented here pose a powerful argument in support of the proposal that AD should be classified as a vascular disorder. According to elementary statistics, the probability or chance that all these findings are due to an indirect pathological effect or to coincidental circumstances related to the disease process of AD seems highly unlikely. The collective data presented in this review strongly support the concept that sporadic AD is a vascular disorder. It is recommended that current clinical management of patients, treatment targets, research designs, and disease prevention efforts need to be critically reassessed and placed in perspective in light of these important findings. PMID- 11935077 TI - Serum gamma-glutamyltransferase as a risk factor of ischemic stroke might be independent of alcohol consumption. PMID- 11935078 TI - Re: Slowly activating potassium conductance (I(D)): a potential target for stroke therapy. PMID- 11935079 TI - Re: Safety of intraventricular sodium nitroprusside and thiosulfate for the treatment of cerebral vasospasm in the intensive care unit setting. PMID- 11935080 TI - Use of hypertonic saline in ischemic stroke. PMID- 11935081 TI - Laterality of ruptured aneurysm has no influence on QT prolongation after subarachnoid hemorrhage. PMID- 11935082 TI - Ambulatory carotid stenting in patients with asymptomatic carotid artery stenosis. PMID- 11935083 TI - Is ambulatory carotid stenting safe? Too early to say. PMID- 11935084 TI - Drugs and recovery: a challenge for a few? PMID- 11935085 TI - Hepatocellular carcinoma: an Asian perspective. AB - Hepatocellular carcinoma (HCC) is one of the most frequently occurring malignancies in Asia. The incidence exceeds 30 cases/100,000/year in the east Asian region. Worldwide, it accounts for almost 1 million deaths/year. The high incidence in Asia is due to the high prevalence of chronic viral hepatitis, mainly chronic hepatitis B. With the introduction of universal vaccination for hepatitis B in some Asian countries in the mid 1980s, some of these countries are experiencing a decline in the incidence of HCC. This probably underscores the point that HCC caused by hepatitis B is a malignancy preventable by vaccine. Due to the relative paucity of symptoms in the early stages and the rapid doubling time of the tumor, most HCCs are discovered late in advanced stages at presentation. Most Asian countries have adopted a screening program for patients at risk. Earlier and smaller HCCs are detected through such programs but these programs have yet to demonstrate improved patient survival. Physicians managing patients with HCC are faced with two main challenges, the malignancy itself and the underlying liver disease. The extent of the tumor and the existing liver function limits the therapeutic choices available. Hepatic resection remains the treatment of choice. However, the majority of patients present with nonresectable tumors. Transarterial chemoembolization, percutaneous ethanol injection and radiofrequency ablation are the other treatment modalities. In patients with small tumors (<5 cm) and poor liver function, liver transplant offers a viable treatment alternative. In summary, the risk factor for HCC in Asia is predominantly chronic hepatitis B. Universal vaccination against hepatitis B is likely to reduce the incidence. The prognosis and outcome of treatment remains poor with a 5-year survival of 35% for patients treated surgically and less than 10% for nonresectable tumors. Current management is aimed at earlier detection and more effective treatment of early HCC. In future, the challenge will be managing HCC in the premalignant stage. PMID- 11935086 TI - Pathology and pathogenesis of hepatocellular carcinoma. AB - Hepatocellular carcinoma (HCC) is the most common primary malignant tumor of the liver and among the most common cancers worldwide. The distribution pattern of HCC shows geographical variation and its pathogenesis is multifactorial. Environmental, infectious, nutritional, metabolic, and endocrine factors contribute directly or indirectly to hepatocarcinogenesis. The synchronous occurrence of different risk factors, such as chronic viral hepatitis B and C, aflatoxin exposure, alcohol consumption or iron overload, in a single patient or patient population further increases the risk. HCC is commonly associated with chronic hepatitis and liver cirrhosis. Different genes have been implicated in the pathogenesis of HCC, and may be divided into four major groups: genes regulating DNA damage response; genes involved in cell cycle control; genes involved in growth inhibition and apoptosis, and genes responsible for cell-cell interaction and signal transduction. Hepatocarcinogenesis is mediated by loss of heterozygosity, somatic mutation, de novo methylation, and/or functional inactivation. As yet, there is no evidence for an ordered sequence of genomic events leading to hepatocarcinogenesis. The pattern of genomic alterations shows great variability, often between two different HCCs from a single patient. HCC evolves from precancerous lesions, and well-differentiated HCC further progresses to a less differentiated form. However, there is still great need for the definition of objective morphological, phenotypic and genetic markers for the progression of HCC. PMID- 11935087 TI - Molecular mechanisms of hepatitis B virus-associated liver cancer. AB - Hepatitis B infection is associated with an increased risk of hepatocellular carcinoma development. Hepatitis B proteins, such as the hepatitis B x protein, the large hepatitis B surface protein, or truncated middle hepatitis B surface proteins, regulate transcription of many candidate genes for liver carcinogenesis by trans-mechanisms. They also alter mechanisms of apoptosis and interfere with nucleotide excision repair of damaged DNA. Together with an influence on cellular signaling, these mechanisms may favor the cell's clonal expansion. PMID- 11935088 TI - Parasitic infections and hepatic neoplasia. AB - Parasitic infections are highly prevalent in the general population. Detecting a relationship between a parasitic infection and cancer is not an easy task; it requires epidemiological, microbiological and molecular biology techniques. In this article the important mechanisms by which parasitic infections may induce carcinogenesis are reviewed. The association between Schistosoma japonicum, Schistosoma mansoni and hepatocellular carcinoma is covered. The cause-and-effect relationship between the liver flukes Opisthorchis viverrini, Opisthorchis felineus, Clonorchis sinensis, and cholangiocarcinoma is also reviewed. The evidence supporting the role of schistosomes and liver flukes in carcinogenesis is discussed. Finally the importance of the primary prevention of these parasitic infections with oncogenic potentials is emphasized. PMID- 11935089 TI - Role of percutaneous ethanol injection in the treatment of hepatocellular carcinoma. AB - In the treatment of early and intermediate hepatocellular carcinoma, the range of indications for percutaneous ablation techniques is becoming wider than surgery or intra-arterial therapies. Indeed, whereas for some years only patients with up to three small tumors were treated, with the introduction of the single-session percutaneous ethanol injection (PEI), performed under general anesthesia, even patients with more advanced disease are now being treated. Although it is understood that partial resection assures the highest local control, the survival rates after surgery are roughly comparable with PEI. The explanation for this is a balance among the advantages and disadvantages of the two therapies. PEI survival curves are better than curves for resected patients who present adverse prognostic factors, and this means that surgery needs a better selection of the patients. Indications for both therapies are reported. An open question remains the choice between PEI and other new ablation procedures. In our department we currently use radiofrequency ablation in the majority of patients but consider PEI and segmental transarterial chemoembolization complementary, and use them according to the features of the disease and the response. PMID- 11935090 TI - Radiofrequency thermal ablation of hepatocellular carcinoma. AB - Hepatocellular carcinoma (HCC) is one of the major malignancies worldwide. Due to advanced or decompensated liver cirrhosis, comorbidity and multicentricity of the tumor lesions, 70-80% of HCC patients are inoperable at the time of diagnosis. Radiofrequency thermal ablation (RFTA) is a new minimally invasive and sage technique for the nonsurgical treatment of HCCs. Similar to other ablation techniques, the treatment strategy depends on several factors, including the patient's clinical status, the stage of liver cirrhosis and of the HCC. RFTA can be performed percutaneously, laparoscopically or after laparotomy. Advanced RFTA equipment, refined techniques of modifying tumor tissue response to RFTA, and combined treatment strategies should lead to better response rates even in larger HCCs. PMID- 11935091 TI - Systemic treatment of hepatocellular carcinoma. AB - Systemic treatment for hepatocellular carcinoma is indicated in locally advanced or metastatic disease. Monochemotherapies have yielded unsatisfactory results with response rates of around 20% but survival is often not improved. Polychemotherapies may induce complete responses but have substantial toxicity and are limited to selected patients with preserved liver function. Hormonal treatment with tamoxifen is ineffective while megestrol has shown an improvement in quality of life. Octreotide can be given even in cases of impaired liver function, has also a favorable side effect profile and can lead to disease stabilization. Adjuvant therapy with interferon is indicated after successful liver resection or transplantation in patients with chronic viral hepatitis, the role of interferon in other indications or in combination with chemotherapy remains to be determined. PMID- 11935092 TI - Gene therapy of hepatocellular carcinoma. AB - The extraordinary versatility of gene therapy opens new possibilities for the treatment of incurable diseases, including hepatocellular carcinoma. Gene therapy strategies against tumors include prodrug activation therapy by the transfer of suicide genes, immunogene therapy, tumoral cell phenotype correction by the inhibition of oncogenes or the transfer of tumor suppressor genes, antiangiogenesis and transfer of oncolytic viruses. The experience accumulated during the last decade of clinical gene therapy indicates that genes can be expressed inside the tumor tissue, but the overall results of the studies conducted so far are still disappointing, mainly due to the poor performance of the currently available gene therapy vectors. This review covers the general aspects of gene therapy vectors, preclinical data available in animal models of hepatocellular carcinoma, and finally a brief summary of the gene therapy clinical trials aimed at the treatment of liver cancer. PMID- 11935093 TI - Current status of liver transplantation for treatment of hepatocellular carcinoma. AB - Hepatocellular carcinoma accounts for more than 5% of all malignancies with a continuous increase worldwide. The most important risk factor is liver cirrhosis, frequently associated with hepatitis B virus or hepatitis C virus infection. Liver resection is the only treatment that can potentially achieve cure. In carefully selected patients with a tumor smaller than 5 cm the 5-year survival is around 50%. The presence of liver cirrhosis and portal hypertension limits the feasibility of hepatic resection. Child-Pugh A patients without major associated risk factors may be considered as the ideal target group for resection. A significant local disease recurrence rate of more than 70% at 5 years is the main problem of hepatic resection. Orthotopic liver transplantation offers the possibility of removing a potentially multicentric tumor and the underlying end stage liver disease. Due to pure selection of suitable candidates the initial reports on the efficacy of liver replacement in a cohort of patients with hepatocellular carcinoma were disappointing. Taking the shortness of donor organs and the high posttransplant tumor recurrence rate into account, several groups developed criteria qualifying transplantation. A tumor size of >6 cm and gross intrahepatic portal vein involvement seem to be of significant prognostic importance. Patients with smaller solitary tumors or less than 3 tumors with a total tumor diameter of <8 cm have the same survival after transplantation as those with benign liver disease. Living donor liver transplantation offers a new approach to overcome the organ shortage and to theoretically extend the indication for transplantation in hepatocellular carcinoma. PMID- 11935094 TI - Prevention of hepatocellular carcinoma in chronic liver disease: molecular markers and clinical implications. AB - The development of hepatocellular carcinoma is generally preceded by chronic liver damage leading to cirrhosis. Prevention of chronic liver diseases can decrease the incidence of hepatic cancer impressively. Many recent investigations have also explored the power of secondary and tertiary prevention in established liver cirrhosis. Screening programs for patients at high risk, antiviral treatment of patients with progressed hepatitis, and adjuvant interventions after curative resection are some of the approaches. However, the cost effectiveness and benefits of such procedures and the prognosis is also dependent on the remaining liver function, there is no consensus to date on how patients should be handled. In the future molecular markers and prognostic scores may help better define the group at risk of developing. To give a perspective to these patients, it is necessary to improve the treatment of hepatocellular carcinoma as well as cirrhosis. PMID- 11935095 TI - Hepatocellular carcinoma in southern Germany: epidemiological and clinicopathological characteristics and risk factors. AB - The aetiology of chronic liver disease leading to hepatocellular carcinoma (HCC) and the clinical characteristics of patients with HCC vary considerably internationally and intranationally. This study analyses the characteristics of HCC patients in southern Germany, a low endemic area of HCC. METHODS: The files of 118 consecutive patients with HCC observed in a single tertiary care hospital between 1994 and 2000 have been reviewed. Epidemiological and clinicopathological characteristics such as age at presentation, ethanol consumption, serological hepatitis virus markers, and fibrosis were studied. Additionally, serum levels of alpha-fetoprotein (AFP) were analysed at the time of diagnosis in 77 patients. RESULTS: The male:female ratio was 4:1 and the mean age at presentation was 61.8 years. Alcohol abuse (49.2%) and chronic hepatitis C infection (17.8%) were the most frequent risk factors. Histologically proven liver cirrhosis in the surrounding non-tumorous tissue was present in only 59.0% of cases. AFP levels were elevated in 78% of cases, but only 34% reached >500 ng/ml, a value considered to be significant for the diagnosis of HCC. AFP levels correlated with the stage of fibrosis. SUMMARY AND CONCLUSIONS: The sensitivity of AFP serum levels as a tumour marker is poor but might help to detect at least a minority of cases. As in other populations within Europe, chronic alcohol abuse is frequently associated with HCC in southern Germany, confirming that alcohol is still the most important risk factor for hepatocarcinogenesis in areas with low hepatitis virus prevalence. Considering the poor prognosis of HCC, prevention is of pivotal importance, particularly for patients with chronic liver disease and other risk factors for the development of HCC. PMID- 11935096 TI - Intracellular accumulation of middle hepatitis B surface protein activates gene transcription. AB - While the natural intact protein does not possess any transactivator function, C terminal truncation of the middle hepatitis B surface (MHBs) protein yields a novel transactivator function. We have previously found that the truncated transactivator protein, MHBs(t167), is not secreted but retained within the secretory pathway. Here, we provide evidence that when full-length MHBs is coexpressed with the truncated MHBs(t167) protein, the secretion of the full length protein is inhibited and both proteins accumulate within the cell. We further show that MHBs, forcibly retained in the cell by C-terminal fusion to the endoplasmic reticulum retention signal KDEL (MHBsKDEL), mimics the effects of MHBs(t167) in enhancing the nuclear-binding activity of transcription factors NFkappaB and AP-1, and activation of NFkappaB- and AP-1-dependent transcription of reporter genes. As is the case for MHBs(t167), MHBsKDEL-dependent activation of NFkappaB is inhibited by the antioxidant N-acetyl-L-cysteine indicating the involvement of reactive oxygen intermediates and suggesting a similar mechanism of activation. This study suggests that the intracellular retention and accumulation of the normally secreted MHBs leads to oxidative stress and activation of transcription. This may be an important but not exclusive mechanism in hepatocarcinogenesis. PMID- 11935097 TI - Prevention and cure of complications from multiple-electrode radiofrequency treatment of liver tumors. AB - BACKGROUND/AIMS: To determine the complication rate of multiple-electrode radiofrequency treatment of liver tumors, and their prevention and cure. METHODS: 114 patients with liver tumors were treated 170 times using multiple-electrode radiofrequency. The clinical complications and their prevention and cure were observed. RESULTS: The complication rate was 9.6%. The complications included 1 case of colon perforation, 4 cases of skin burn, 5 cases of hydrothorax, and 1 case subcutaneous hemorrhage, but none of the patient died due to these complications. CONCLUSION: The complication rate of multiple-electrode radiofrequency treatment of liver tumors is low and the complications could be prevented and cured. PMID- 11935099 TI - Point of view. PMID- 11935098 TI - Effects of training on female trapezius Myalgia: An intervention study with a 3 year follow-up period. AB - STUDY DESIGN: A prospective randomized clinical trial was conducted. OBJECTIVE: To evaluate the long-term effects of three training interventions on women with work-related trapezius myalgia. SUMMARY OF BACKGROUND DATA: Studies with long term follow-up evaluation of interventions for neck pain are scarce and usually cover fewer than 12 months. METHODS: For this study, 126 women with work-related trapezius myalgia were randomized into strength, muscular endurance, or coordination training or into a nontraining reference group. Intervention training occurred three times a week for 10 weeks. Assessment of pain intensity, pressure pain thresholds, symptom frequency, perceived health, and work and exercise habits was performed before and immediately after interventions, then at clinical examinations 8 months and 17 months after the interventions. Participation rates at these follow-up assessments were 84% and 81%. At 3 years after the interventions, a questionnaire was answered by 94% of the participants, 17 of whom were dropouts that never participated in any of the four intervention groups. RESULTS: All the training programs showed similar pain reducing effects immediately after the interventions. Pain reductions were maintained at follow-up evaluations, but at the 8-month follow-up assessment, there were no differences between the training groups and the reference group on any variable. At 3 years after the interventions, the intervention groups did not differ from the dropout group. Almost half (49%) of the women had persistent symptoms at the 3-year follow-up assessment. CONCLUSIONS: The long-term effect of all the training programs was low. Pain in neck and shoulder muscles was persistent in a large proportion of the women over the 3 years. PMID- 11935100 TI - Ultrastructure of the human intervertebral disc during aging and degeneration: comparison of surgical and control specimens. AB - STUDY DESIGN: Human intervertebral disc tissue from the annulus was obtained in a prospective study investigating the ultrastructural features of disc cells and extracellular matrix. Experimental studies were approved by the authors' Human Subjects Institutional Review Board. Discs were obtained from surgical specimens and control donors. OBJECTIVE: To compare the cellular and extracellular matrix characteristics of the annulus from control and surgical disc specimens using electron microscopy and specialized fixation that visualizes proteoglycans. SUMMARY OF THE BACKGROUND DATA: The ultrastructural features of disc cells and the disc matrix have received little attention, as compared with the literature on age- and disease-related changes in bone and cartilage. METHODS: Ultrastructural studies investigated disc tissue obtained from control and surgical disc specimens using transmission electron microscopy. Specialized fixation with ruthenium red was used to highlight matrix proteoglycans. RESULTS: Cellular and extracellular matrix fine structure was assessed in disc specimens from 29 control donors (newborns to 79-year-olds) and surgical disc specimens from 49 patients (16- to 77-year-olds). Control and surgical tissue showed similar ultrastructural features. Unusual matrix surrounding and encircling single cells or clusters of cells was common (48% of control and 63% of surgical specimens) and often contained fibrous long-spacing collagen (41.3% of control and 36.7% of surgical specimens). Ruthenium red greatly aided visualization of proteoglycans pooled in lacunar spaces. Variable cross-sectional diameters of collagen fibrils was present in 34% of control and 59% of surgical specimens. Regions with sparse interterritorial matrix were common. Cell morphology showed both cells with apoptotic nuclei and synthetically active cells that appeared healthy. CONCLUSIONS: Control and surgical specimens of the annulus showed similar ultrastructural features. Heterogeneity of collagen fibril diameter is an important observation because it is believed that fibril size relates to biomechanical disc function. Fibrous long-spacing collagen may reflect extracellular matrix remodeling or the presence of previous fibril depolymerization followed by repolymerization and reassociation with proteoglycans. Synthetic activity of disc cells is reflected in active rough endoplasmic reticulum, Golgi, and pools of proteoglycans in lacunar spaces and unusual extracellular matrix components that encircle cells and cell clusters. Such components may influence biomechanical quality. Departures from normal extracellular matrix organization of the aging or degenerating disc undoubtedly contribute to decreased biomechanical function of the annulus because they disrupt the normal annulus architecture. This study underscores the need for a fuller understanding of the dynamic relation between disc cells and the surrounding extracellular matrix, which they continually produce and remodel. PMID- 11935101 TI - Advantage of pedicle screw fixation directed into the apex of the sacral promontory over bicortical fixation: a biomechanical analysis. AB - STUDY DESIGN: A biomechanical study of human cadaveric sacra using insertional torque and bone mineral density was conducted to determine the optimal sagittal trajectory of S1 pedicle screws. OBJECTIVE: To measure the maximal insertional torque of sacral promontory versus bicortical pedicle screw fixation. SUMMARY OF BACKGROUND DATA: Fixation of instrumentation to the sacrum is commonly accomplished using S1 pedicle screws, with previous studies reporting biomechanical advantages of bicortical over unicortical S1 screws. The biomechanical effect of bicortical screws (paralleling the endplate) versus screws directed into the apex of the sacral promontory is unknown. METHODS: For this study, 10 fresh frozen cadaver sacra were harvested and evaluated with dual energy radiograph absorptiometry to assess bone mineral density. Matched 7.5-mm monoaxial stainless steel pedicle screws then were randomly assigned by side (left versus right) and placed bicortically or into the apex of the sacral promontory under direct visualization. Maximum insertional torque was recorded for each screw revolution with a digital torque wrench (TQJE1500, Snap-On Tools, Kenosha, WI). RESULTS: Maximum bicortical S1 screw insertional torque averaged 5.22 +/- 0.83 inch-pounds, as compared with the maximum sacral promontory S1 screw insertional torque of 10.34 +/- 1.94 inch-pounds. This resulted in a 99% increase in maximum insertional torque (P = 0.005) using the "tricortical" technique, with the screw directed into the sacral promontory. Mean bone mineral density was 940 +/- 0.25 mg/cm2 (range, 507-1428 mg/cm2). The bone mineral density correlated with maximal insertional torque for the sacral promontory technique (r = 0.806; P = 0.005), but not for the bicortical technique (r = 0.48; P = 0.16). CONCLUSIONS: The screws directed into the apex of the sacral promontory of the S1 pedicle resulted in an average 99% increase in peak insertional torque (P = 0.005), as compared with bicortical S1 pedicle screw fixation. Tricortical pedicle screw fixation correlates directly with bone mineral density. PMID- 11935102 TI - Validity study for the cervical range of motion device used for lateral flexion in patients with neck pain. AB - STUDY DESIGN: A set of measurements was compared with an accepted gold standard using a methodologic design. OBJECTIVE: To estimate the criterion validity of the cervical range of motion device used for lateral flexion in patients with neck pain. SUMMARY OF BACKGROUND DATA: Reliability of the cervical range of motion device has been well established. At this writing, only validity for flexion and extension of the cervical spine has been investigated with this device. METHODS: The sample consisted of 24 volunteer subjects who had previously received physiotherapy for neck pain. In the radiograph department, subjects were stabilized on a chair. A first reading on the cervical range of motion device and a radiograph were taken in a neutral starting position. The subject then was asked to perform a maximal right lateral flexion, which was followed by a reading of the device and a second radiograph. The same procedures were followed for left lateral flexion. RESULTS: The cervical range of motion device demonstrated a very good linear relation with the radiograph measurements: left lateral flexion (r = 0.82,; 95% confidence interval, 0.62-0.92), right lateral flexion (r = 0.84; 95% confidence interval, 0.66-0.93). CONCLUSIONS: The cervical range of motion device showed very good validity for measurement of lateral flexion in this population of patients with neck pain. Because the reliability of the cervical range of motion device was established previously, the results of this study suggest that the device be used as an outcome measure for the cervical flexion, and for flexion and extension. PMID- 11935103 TI - Factors predicting hospital stay, operative time, blood loss, and transfusion in patients undergoing revision posterior lumbar spine decompression, fusion, and segmental instrumentation. AB - STUDY DESIGN: A retrospective chart review was conducted for 112 patients who underwent revision posterior lumbar spine decompression, fusion, and segmental instrumentation. OBJECTIVE: To ascertain factors predicting hospital stay, operative time, blood loss, and transfusion in patients undergoing revision posterior lumbar spine decompression, fusion, and segmental instrumentation. SUMMARY OF BACKGROUND DATA: Posterior lumbar spine decompression and fusion with segmental instrumentation is a common procedure in the treatment of degenerative lumbar spine disorders. Many patients undergoing this procedure have had previous lumbar spine surgery, yet little is known about the factors predicting hospital stay, operative time, blood loss, and transfusion. METHODS: The charts of 112 patients (53 men and 59 women) with degenerative lumbar spinal stenosis who underwent revision surgery from March 1992 to June 1999 were reviewed. Their average age was 54 years (range, 27-84 years). All the surgeries included decompression and fusion with segmental instrumentation. The patients' demographics, comorbid conditions, factors related to previous lumbar spine surgery, diagnosis, number of levels fused, and preoperative hemoglobin and hematocrit were collected and used as the independent variables. Multiple regression analysis was used to ascertain factors predicting length of hospital stay, operative time, intraoperative blood loss, and transfusion. RESULTS: The mean length of hospital stay was 6 +/- 2.4 days, the operative time 280 +/- 62 minutes, the estimated intraoperative blood loss 1073 +/- 716 mL, and the total volume of blood transfused 1.04 +/- 1.17 U. For 63% of the patients, a blood transfusion was needed. Increasing age was the significant predictor for hospital stay (P < 0.001). The factors predicting operative time were number of levels fused (P < 0.001), diagnosis of degenerative scoliosis (P < 0.05), and excessive body weight (P < 0.01). The factors predicting intraoperative blood loss were number of levels fused (P < 0.01), body weight (P < 0.001), and high preoperative hemoglobin (P < 0.001). Both logistic and linear regression analysis showed that the factors predicting blood transfusion were number of levels fused (P < 0.01), age (P < 0.05), and low preoperative hemoglobin (P < 0.001). Other factors associated with hospital stay and blood transfusion were unemployment associated with three or more comorbid conditions and complications. The women had less intraoperative blood loss (P < 0.01), but received more transfused blood than the men (P < 0.01). CONCLUSIONS: Number of levels fused and age seem to be the most significant factors predicting hospital stay, operative time, intraoperative blood loss, and transfusion in patients undergoing revision posterior lumbar spine decompression, fusion, and segmental instrumentation. PMID- 11935104 TI - Factors related to false- versus true-positive neuromonitoring changes in adolescent idiopathic scoliosis surgery. AB - STUDY DESIGN: A retrospective study of 134 adolescent patients who underwent surgical correction of idiopathic scoliosis between June 1992 and August 1998 was conducted. OBJECTIVE: To examine factors related to changes in somatosensory evoked potentials with or without neurogenic motor-evoked potentials. SUMMARY OF BACKGROUND DATA: Studies document and demonstrate threshold criteria for changes in neuromonitoring that predict changes in spinal cord function. Rates of false negative occurrences are low, yet higher rates of false-positive findings may result. METHODS: All the patients had somatosensory monitoring, and 71 patients had both somatosensory-evoked potential and neurogenic motor-evoked potential monitoring. Gender, age, curve types, duration of surgery, type and amount of instrumentation, and amount of correction were examined for their effects on monitoring. Estimated blood volume loss as well as high and low mean arterial pressure and its variance were assessed at the start, middle, and conclusion of the procedure. RESULTS: According to the findings, 122 patients (91%) had no monitoring changes and no postoperative neurologic deficit. Six patients (4.5%) had false-positive readings. Six patients had a postoperative motor or sensory deficit, all of which resolved within 18 months. False-positive readings were associated with greater variability in mean arterial pressure. No consistent predictions could be made about the incidence of cord injury if neuromonitoring changes returned to baseline before the end of surgery. CONCLUSIONS: Questions remain about the predictive accuracy of somatosensory-evoked and neurogenic motor evoked potentials. According to the findings in this study, in which there were no false-negative readings and a modest false-positive rate, continued use of these methods is recommended. Higher false-positive rates were seen in patients with greater lability in mean arterial pressure. A wake-up test is recommended for all cases in which threshold monitoring changes occur because cases of spinal cord injury may exist even when monitored variables return to baseline. PMID- 11935105 TI - Radiographic classification of L5 isthmic spondylolisthesis as adolescent or adult vertebral slip. AB - STUDY DESIGN: A radiographic and morphologic study was conducted to investigate low-grade spondylolisthesis in cases with preexisting isthmic spondylolysis of L5. OBJECTIVE: To distinguish radiographically between vertebral slips before and after skeletal maturity as determined by deformities of the sacral endplate. SUMMARY AND BACKGROUND DATA: Very few reports have shown that spondylolisthesis with preexisting isthmic defects of L5 develops frequently in adulthood. The prognostic factors of the vertebral slip have remained unclear. It is hard to determine the onset time of low-grade spondylolisthesis. METHODS: This study examined plain radiographs of 367 adult patients with pars defects of L5 (213 without slippage and 154 with Grade 1 or 2 spondylolisthesis) and 310 control subjects, ages 20 to 59 years at the first visit. The following parameters were measured and analyzed for each age decade: the sacral table index (anteroposterior width of the sacral endplate expressed as a percentage of the anteroposterior diameter of the upper L5 endplate), the sacral table angle (formed by the sacral endplate with the posterior wall of S1), the relative thickness of the L5 transverse process, and the iliac crest height. RESULTS: The prevalence of patients with slippage who met deformity criteria (sacral table index > 102% [the mean plus 2 standard deviations of the controls] and sacral table angle /=89 degrees [mean minus 2 standard deviations of the controls]) was 0% in the third decade, but increased remarkably in the fifth and sixth decades. Of the 213 patients without slippage, 8 patients in whom new slippage developed during long term follow-up evaluation all had a normally-shaped sacral table. The prevalence of patients without slippage decreased gradually with age, and elderly patients had relatively broader transverse processes and a higher iliac crest line. CONCLUSIONS: The authors considered that the slips with and those without deformities of the sacral table had developed in adolescence and adulthood, respectively. Using new radiographic parameters that indicate widening and tilting of the sacral table, low-grade isthmic spondylolis thesis can be categorized into "adolescent and adult vertebral slips." PMID- 11935106 TI - Gadopentetate dimeglumine as an intradiscal contrast agent. AB - STUDY DESIGN: Magnetic resonance discography using gadolinium as an intradiscal contrast agent was investigated in a prospective study. OBJECTIVE: To evaluate the utility of magnetic resonance discography using gadolinium as an intradiscal contrast agent, as compared with the current standard of computed tomographic discography using intradiscal iodinated contrast agent. SUMMARY OF BACKGROUND DATA: Magnetic resonance arthrography with gadolinium has been used to evaluate shoulder, elbow, wrist, hip, knee, and other joints. Gadolinium has not been used as an intradiscal contrast agent. The authors' preliminary results using gadolinium as an intradiscal contrast agent for magnetic resonance discography were reported previously. They report the results of their completed study. METHODS: For this study, 42 disc levels were studied in 13 patients. Water soluble iodinated contrast and gadopentetate dimeglumine were injected at each disc level. After discography, anteroposterior and lateral radiographs, computed tomography scans, and T1-weighted magnetic resonance imaging were performed. Six physicians interpreted the results from each disc level in a blinded fashion. Interscan and interobserver interpretation variability was determined for magnetic resonance discography and computed tomographic discography using the Pearson correlation-coefficient (r) test. RESULTS: Interscan variability was highly correlated between computed tomographic discography and magnetic resonance discography when used to determine disc normality (r = 0.87), general degeneration (r = 0.87), anular fissure (r = 0.89), disc herniation (r = 0.92), and contrast leakage (r = 0.77). Interscan variability was assessed by the Pearson test, and all values of r for all the readers were noted to be statistically significant at P values less than 0.01. Interobserver variability was significantly correlated among the four more experienced readers (neuroradiologists and spine surgeons), but not among the spine fellows. Interobserver variability was also significantly correlated by computed tomographic (CT) and magnetic resonance imaging (MRI) discography for disc normality (CT r = 0.60; MRI r = 0.56), general degeneration (CT r = 0.76; MRI r = 0.71), anular fissure (CT r = 0.79; MRI r = 0.84), and disc herniation (CT r = 0.63; MRI r = 0.64). The readings for contrast leakage did not reach statistical significance for computed tomographic or magnetic resonance discography. CONCLUSIONS: The high interscan and interobserver correlation rates obtained for magnetic resonance discography using gadolinium, as compared with the standard computed tomographic discography technique, indicate that magnetic resonance discography may be an acceptable substitute for the imaging of disc pathology. Magnetic resonance discography with gadolinium can be recommended for patients allergic to iodinated contrast agents and for patients who wish to limit their radiation exposure. PMID- 11935107 TI - Utility of three-dimensional and multiplanar reformatted computed tomography for evaluation of pediatric congenital spine abnormalities. AB - STUDY DESIGN: A retrospective radiographic review of 31 patients with congenital spine abnormalities who underwent conventional radiography and advanced imaging studies was conducted. OBJECTIVE: To analyze the utility of three-dimensional computed tomography with multiplanar reformatted images for congenital spine anomalies, as compared with plain radiographs and axial two-dimensional computed tomography imaging. SUMMARY OF BACKGROUND DATA: Conventional radiographic imaging for congenital spine disorders often are difficult to interpret because of the patient's small size, the complexity of the disorder, a deformity not in the plane of the radiographs, superimposed structures, and difficulty in forming a mental three-dimensional image. Multiplanar reformatted and three-dimensional computed tomographic imaging offers many potential advantages for defining congenital spine anomalies including visualization of the deformity in any plane, from any angle, with the overlying structures subtracted. METHODS: The imaging studies of patients who had undergone a three-dimensional computed tomography for congenital deformities of the spine between 1992 and 1998 were reviewed (31 cases). All plain radiographs and axial two-dimensional computed tomography images performed before the three-dimensional computed tomography were reviewed and the findings documented. This was repeated for the three-dimensional reconstructions and, when available, the multiplanar reformatted images (15 cases). In each case, the utility of the advanced imaging was graded as one of the following: Grade A (substantial new information obtained), Grade B (confirmatory with improved visualization and understanding of the deformity), and Grade C (no added useful information obtained). RESULTS: In 17 of 31 cases, the multiplanar reformatted and three-dimensional images allowed identification of unrecognized malformations. In nine additional cases, the advanced imaging was helpful in better visualizing and understanding previously identified deformities. In five cases, no new information was gained. The standard and curved multiplanar reformatted images were best for defining the occiput-C1-C2 anatomy and the extent of segmentation defects. The curved multiplanar reformatted images were especially helpful in keeping the spine from "coming in" and "going out" of the plane of the image when there was significant spine deformity in the sagittal or coronal plane. The three-dimensional reconstructions proved valuable in defining failures of formation. CONCLUSIONS: Advanced computed tomography imaging (three-dimensional computed tomography and curved/standard multiplanar reformatted images) allows better definition of congenital spine anomalies. More than 50% of the cases showed additional abnormalities not appreciated on plain radiographs or axial two-dimensional computed tomography images. Curved multiplanar reformatted images allowed imaging in the coronal and sagittal planes of the entire deformity. PMID- 11935108 TI - Discriminative and predictive validity assessment of the quebec task force classification. AB - STUDY DESIGN: A prospective cohort study of workers with low back pain who had been absent from work for more than 4 weeks was conducted. OBJECTIVE: To assess the discriminative and predictive validity of the Quebec Task Force Classification for workers during the subacute phase of disability from back pain. SUMMARY OF BACKGROUND DATA: The Quebec Task Force Classification was designed for clinical decision making, prognosis establishment, quality of care evaluation, and scientific research in low back pain. METHODS: For this study, 104 workers absent from work because of back pain were classified according to the first four categories of the Quebec Task Force Classification 4 weeks after their first day of work absence. They then were randomized into four treatment groups: standard care (control), clinical-rehabilitation intervention, occupational intervention, and the Sherbrooke model (a combination of the clinical-rehabilitation and occupational interventions). Functional status, pain level, and work status were assessed at baseline and after 1 year. Duration of full compensation and back-related costs were calculated over a mean follow-up period of 6.5 years. The discriminative validity of the Quebec Task Force Classification was evaluated using Kendall tau correlation coefficients. Predictive validity was evaluated using logistic regression analyses. Age, gender, comorbidities, body mass index, and treatment group were considered as potential confounders. RESULTS: Significant but low correlation coefficients were found between Quebec Task Force Classification categories and functional status scores at baseline. Subjects classified as having distal radiating pain (categories 3 and 4) at baseline were more likely to have a lower functional status, higher pain level, and no return to regular work at the 1-year follow-up evaluation. They also were more likely to accumulate more days of full compensation and to cost more after a mean follow-up period of 6.5 years. CONCLUSION: The Quebec Task Force Classification demonstrated good predictive ability by discriminating between subjects with and those without distal radiating pain. PMID- 11935110 TI - Point of view. PMID- 11935109 TI - Interobserver and intraobserver reliability of Lenke's new scoliosis classification system. AB - STUDY DESIGN: The assignment of idiopathic scoliosis curves to the curve types,1 6 to the lumbar spine modifier (A, B, or C), and to the sagittal thoracic modifier (-, N, +), as recently described by Lenke et al, was evaluated by five observers on two occasions. OBJECTIVE: To determine the intraobserver and interobserver reliabilities of Lenke's new system for classifying idiopathic scoliosis. SUMMARY OF BACKGROUND DATA: Lenke et al recently introduced a new system for classifying idiopathic scoliosis. METHODS: Preoperative standing long cassette coronal and sagittal radiographs as well as side-bending radiographs of 51 consecutive patients who underwent surgery for idiopathic scoliosis were labeled with the Cobb angles for all the curves. The center sacral vertical line was marked as the bisection of the proximal sacrum perpendicular to the true horizontal line. Five observers independently assigned a curve type, a lumbar spine modifier, and a sagittal thoracic modifier to each curve following the guidelines as described by Lenke et al. Assignment of the curves was repeated 3 weeks later, with the curves presented in a different order. Kappa coefficients were used to determine the interobserver and intraobserver reliabilities. RESULTS: All five reviewers agreed on the overall classification in 21 (41%) of the 51 patients. A mean kappa value of 0.62 was determined for interobserver reliability, and a mean kappa of 0.73 for intraobserver reliability. Determination of an upper thoracic curve as structural or nonstructural and assignment of a lumbar spine modifier were the main reasons for disagreement. CONCLUSIONS: Lenke's new classification system is more reliable than the older King classification, but proper classification of high thoracic and lumbar curves seems to be difficult. PMID- 11935111 TI - Work-related outcomes in occupational low back pain: a multidimensional analysis. AB - OBJECTIVES: This pilot study explored a broad range of work-related outcomes for occupational low back injuries. METHODS: A model of occupational outcomes and a survey instrument were developed on the basis of interviews, expert opinion, and literature reviews. New Hampshire workers who had an occupational back injury a year before the study were sampled from first reports of injury and sent a mailed survey about their postinjury experiences and related factors. RESULTS: Of 251 randomly selected cases, a valid address could be identified for 121, and 99 patients responded. Almost 60% of the respondents had lost 1 week of work or more. At 1 year after injury, half of the respondents had returned to their preinjury job and employer, and 20% were unemployed, half of them because of the injury. Most working respondents reported no decrease in their work capacity. However, 68% still had pain exacerbated by work, and 47% worried that their condition would worsen with continued work. Reinjury occurred in 42% of the respondents. The work-related outcome measures were largely independent of each other. Exploratory multivariate analyses demonstrated unique patterns of factors associated with each outcome. Reinjury risk was significantly greater in respondents whose employers offered accommodations or whose postinjury jobs had greater ergonomic risk. The small sample size limited the ability to achieve statistically significant results in multivariate analyses. CONCLUSIONS: Simply measuring return to work did not appear to capture the full range of job-related consequences from occupational back injuries in this pilot evaluation. Timing of return to work, occupational ergonomic risks, and appropriate job modifications appeared to be particularly important in a safe return to the job after an occupational low back injury. Results suggest opportunities to address risk factors that may improve work outcomes. PMID- 11935112 TI - Video-assisted thoracoscopic surgery for thoracic disc disease: Classification and outcome study of 100 consecutive cases with a 2-year minimum follow-up period. AB - STUDY DESIGN: Prospectively collected data from regular clinical follow-up evaluations were tabulated, analyzed, reviewed using a patient self-reported questionnaire. OBJECTIVE: To develop a classification system and present the long term functional outcome of video-assisted thorascopic surgery for refractory thoracic disc disease. SUMMARY OF BACKGROUND DATA: Recent studies have found an 11.1% to 14.5% prevalence of thoracic disc herniations. Surgical approaches have included laminectomy, pediculectomy, costotransversectomy, lateral extracavitary, transverse arthropediculectomy, transthoracic-transpleural thoracotomy, and thoracoscopy. Recent reports have documented encouraging early results with video assisted thorascopic surgery for thoracic disc herniations. Comparisons between thoracoscopy and open thoracotomy have demonstrated improvement in postoperative pain and morbidity with the use of endoscopic techniques. METHODS: This study included 100 consecutive patients (45 women and 55 men) with an average follow-up evaluation of 4 years (range, 2-6 years). The average age of the patients was 42 years (range, 22-76 years). The average duration of symptoms was 26 months (range, 6-96 months), and 18 patients had undergone prior spine surgery. Patients were graded as follows according to the presenting symptoms (Table 1): Grade 1 (pure axial; n = 28), Grade 2 (pure radicular; n = 5), Grade 3A (axial and thoracic radicular; n = 38), Grade 3B (axial with lower leg pain; n = 19), Grade 4 (myelopathic; n = 8), or Grade 5 (paralytic = 2). RESULTS: A total of 117 discs were excised in 100 patients. Of the 40 patients who underwent fusion, 27 had autologous rib struts and 13 had threaded fusion. The mean operative time was 173 minutes, blood loss 259 mL, average ICU stay less than 1 day, and average hospital stay 4 days. Minor complications occurred in 21 patients, all of which resolved with no untoward effect. No patient's neurologic status worsened. Four patients underwent a secondary fusion, and a pseudarthrosis developed in one patient. Clinical success was defined objectively as an improvement in Oswestry score of 20% or more at 2 years and at final follow-up assessment, as compared with the preoperative Oswestry score. Overall, objective clinical success was observed at 2 years in 73% of the patients, and at final follow-up assessment in 70% of patients. The average percentage of improvement in Oswestry scores was most marked in Grade 4 patients (myelopathy; 60%), followed by Grade 3A patients (axial and thoracic radicular pain; 37%), Grade 3B patients (axial with leg pain; 28%), and Grade 1 patients (pure axial; 24%). The Oswestry disability score (Table 2) and back pain visual analog score (Table 3) also were significantly improved (P < 0.05) at 2 years and at final follow-up assessment in these patients. In the Grade 2 patients, those pure thoracic radicular pain, Oswestry scores initially improved significantly up to 1 year (P < 0.05). At 2 years, no significant improvement could be shown, and four of the five Grade 2 patients reported increased axial pain as their main symptom at the final follow-up assessment. Significant improvement also was seen in patients with no prior spine surgery and patients with preoperative Oswestry disability scores greater than 50. Of the 68 patients who responded to the final questionnaire, 12 rated the procedure as excellent, 37 as good, 11 as fair, and 8 as poor. Also, 57 (83.8%) of these 68 patients were satisfied and indicated they would recommend the surgery. Of the 36 patients at the final follow up assessment who had severe disability, 34 (94%) were satisfied, as compared with 23 of the 32 patients (72%) who had presented with milder disability. CONCLUSIONS: The clinical classification system helps in differentiating different presentations of thoracic disc disease and their final outcome. Video-assisted thorascopic surgery appears to be a safe and efficacious method for the treatment of refractory symptomatic thoracic disc herniations. The current data suggest that the procedure has an acceptable long-term outcome, with an 84% overall subjective patient satisfaction rate, and with objective long-term clinical success achieved in 70% of patients. PMID- 11935113 TI - Point of view. PMID- 11935115 TI - Olfactory ensheathing cells: their potential use for repairing the injured spinal cord. AB - STUDY DESIGN: The literature concerning the potential use of olfactory ensheathing cells for repairing damaged spinal cord was reviewed. OBJECTIVE: To engender a better understanding of the role that olfactory ensheathing cells play in spinal cord regeneration. SUMMARY OF BACKGROUND DATA: Intraspinal transplants (e.g., fetal neuronal cells, progenitor stem cells, and olfactory ensheathing cells) have been used to restore intraspinal circuitry or to serve as a "bridge" for damaged axons. Among these transplants, olfactory ensheathing cells provide a particularly favorable substrate for spinal axonal regeneration because these cells can secrete extracellular molecules and neurotrophic factors and have the ability to migrate into gliotic scar tissue, an important attribute that might be associated with high potential for axonal regeneration. METHODS: Recent advances using centrally and peripherally derived olfactory ensheathing cells to promote spinal cord regeneration were reviewed. RESULTS: Both centrally and peripherally derived olfactory ensheathing cells can lead to a degree of functional and anatomic recovery after spinal cord injury in adult animals. CONCLUSION: Olfactory ensheathing cells from olfactory lamina propria in the nose are among the best transplants for "bridging" descending and ascending pathways in damaged spinal cord. PMID- 11935114 TI - Thoracoscopic discectomy and fusion in an animal model: safe and effective when segmental blood vessels are spared. AB - STUDY DESIGN: Disc-endplate excision and spine fusion were compared in animals randomly assigned to segmental vessel-spared and segmental vessel-ligated groups in an in vivo goat model of anterior spine discectomy and fusion using thoracoscopic techniques. OBJECTIVES: To compare safety and completeness of disc and endplate excision, and to perform a histologic and biomechanical comparison between fusion masses when the segmental vessels are spared and when they are ligated using thoracoscopic techniques. SUMMARY OFF BACKGROUND DATA: Because thoracoscopy is relatively new and technically demanding, many surgeons ligate the segmental blood vessels to enhance spine exposure and limit the risk of injury during discectomy and fusion. Although rare, spinal cord compromise secondary to segmental vessel ligation has been reported. METHODS: This study was divided into two phases. In Phase 1, 10 mature goats were randomly assigned to either the segmental vessel-ligated or the segmental vessel-spared group. Disc and endplate excision was performed at six consecutive thoracic levels in each animal (30 levels per group). The animals were killed, and the depth of disc excision was measured in the transverse and sagittal planes. The vertebral bodies then were separated through the disc space; photographic images of the endplates were digitized, and the area of endplate excision was calculated. In Phase 2, 12 mature goats were randomly assigned to the segmental vessel-ligated or vessel spared group, and five noncontiguous thoracic segments were fused using autologous iliac crest graft. At 4 months the animals were killed, and the spines were harvested. At each disc level, the three-dimensional rotational and translational motions were analyzed and histomorphometric analysis was performed. RESULTS: Phase 1: Each animal survived the operative procedure, and no surgical complications occurred. No difference was found between vessel-ligated and vessel spared groups in operative time (21.8 vs 22.7 minutes per disc), blood loss (97 vs 159 mL), or transverse (81% vs 74%) or sagittal (85% vs 85%) disc excision. The total area of endplate excision was 70% in the vessel-ligated group and 67% in the vessel-spared group (P > 0.1). Phase 2: Biomechanical testing demonstrated no difference in stiffness of the fused segments between the two groups in flexion-extension or axial rotation. However, greater flexibility in lateral bending was found in the specimens whose vessels were ligated (P < 0.05). The percentage of trabecular bone volume was similar between the two groups. CONCLUSIONS: The segmental vessels in the thoracic spine can be effectively spared without injury during disc excision and fusion. Although slightly more disc area was excised with ligation of the vessels, this was not statistically significant, and the fusion mass was similar between the two groups. Sparing the segmental vessels may provide blood supply that aids fusion mass, and the result may be greater spine stiffness in the coronal plane. Sparing the segmental vessels during thoracoscopic anterior disc excision and fusion can be safe. It should be considered in patients with a high risk for neurologic injury because of decreased spinal cord perfusion in revision surgery, severe kyphosis, congenital anomalies. Because the neurologic risk of vessel ligation has not been clearly established for idiopathic scoliosis, the surgeon will have to consider the risk-benefit ratio of adopting these methods when deciding not to ligate vessels in these patients. PMID- 11935116 TI - Sacral origin of a spinal dural arteriovenous fistula: case report and review. AB - STUDY DESIGN: A case of spinal dural arteriovenous fistula arising from a branch of the internal iliac artery is reported. OBJECTIVE: To report a rare case of spinal dural arteriovenous fistula supplied by a lateral sacral artery and treated with endovascular therapy. SUMMARY OF BACKGROUND DATA: Spinal dural arteriovenous fistulas usually occur in the thoracic and lumbosacral regions and arise from the intercostal and lumbar arteries. Rarely, they may occur in the sacral region, as in the reported case. METHODS: A 60-year-old man presented with progressive lower extremity paresis and decreased sensation below the waist of 6 months duration, which had progressed to paraparesis. Diagnostic imaging included magnetic resonance imaging and spinal angiography. RESULTS: A sacral spinal dural arteriovenous fistula was diagnosed with spinal angiography, which showed the spinal dural arteriovenous fistula arising from the right lateral sacral artery branches at S2, and magnetic resonance imaging, which showed enlarged pial vessels along the surface of the spinal cord and central cord hyperintensity, with peripheral hypointensity on T2-weighted images. The patient was definitively treated with endovascular therapy using polyvinyl alcohol particles and Tornado coils. His symptoms almost completely resolved within 6 months of therapy. CONCLUSIONS: Although surgical ligation is the treatment of choice, endovascular therapy may be an effective treatment for patients with sacral region spinal dural arteriovenous fistula in cases of high surgical risk. Spinal angiography remains the definitive diagnostic examination for pinpointing the site of the dural arteriovenous fistula. PMID- 11935117 TI - Letters. PMID- 11935118 TI - Letter to the editor. PMID- 11935119 TI - Comparative morphometry of L4 vertebrae: comparison of large animal models for the human lumbar spine. AB - STUDY DESIGN: Anatomic analysis of L4 vertebral morphometry comparing specimens harvested from humans and five common large animal species. OBJECTIVE: To compare fundamental structural similarities and differences in the vertebral bodies of commonly used experimental animals relative to human vertebrae. SUMMARY OF BACKGROUND DATA: Animal models are commonly used for assessment of spine fusion, instrumentation techniques, and vertebral bone biology. Among the animals used, the lumbar vertebrae exhibit considerable anatomic variability. The goal of this study was to determine which of the animals commonly used for spine research is best suited as an anatomic model for the human lumbar spine. METHODS: Morphometric features of the L4 vertebrae of five common research animals were compared with those of the human L4 vertebrae. Mature canines, immature pigs, mature micropigs, mature dairy goats, and mature sheep were analyzed. These species were chosen because they are commonly selected research animals, and most research facilities do not need to be modified to use them. The samples included ten L4 vertebrae of each animal species and seven human L4 vertebrae. Each specimen was meticulously cleaned of all soft tissue. The measurements were grouped into vertebral body parameters, neural canal dimensions, and pedicle and facet morphometery. The mean of each anatomic measurement was compared using a single factor analysis of variance and a Scheffe's post hoc test, with 0.05 denoting significance. RESULTS: The human vertebral body was significantly wider and deeper in the anteroposterior plane than any of the animals studied. However, the mean vertebral body height of the sheep and goat significantly exceeded that of the human specimens. The mean pedicle angle of every animal species was significantly greater than that of the human. The mean pedicle width of the micropig and goat were significantly narrower than the human pedicles, and the dog specimens lacked a definable pedicle altogether. There was no significant difference in mean pedicle width between any of the remaining species and the human specimens. Facet tropism and radius of curvature of the sheep and goat specimens differed significantly from the remaining selections. CONCLUSIONS: When posterior pedicle instrumentation is part of a testing protocol, the increased pedicle angle and lack of vertebral body depth found in all animals studied must be kept in mind. In addition, when testing interbody cages designed to stabilize the spine and promote fusion, one must be aware of the decreased vertebral body depth and width in these animals, as compared with humans. Physeal defects in the immature pig may alter specific biomechanical results during failure or fatigue testing, or in basic studies of vertebral bone material properties. In all cases, instrumentation and hardware must be sized appropriately to the selected model to provide meaningful results. PMID- 11935120 TI - Interexaminer reliability of low back pain assessment using the McKenzie method. AB - STUDY DESIGN: A test-retest design was used. OBJECTIVE: To assess interexaminer reliability of the McKenzie method for performing clinical tests and classifying patients with low back pain. SUMMARY OF BACKGROUND DATA: Clinical methods and tests classifying patients with nonspecific low back pain have been based mainly on symptom duration or extent of pain referral. The McKenzie mechanical diagnostic and classification approach is a widely used noninvasive, low technology method of assessing patients with low back pain. However, little is known about the interexaminer reliability of the method, previous studies having yielded conflicting results. METHODS: For this study, 39 volunteers with low back pain, mean age 40 years (range, 24-55 years), were blindly assessed by two physical therapists trained in the McKenzie method. The variability of two examiners for binary decisions was expressed by the kappa coefficient, and by the proportion of observed agreement, as calculated from a 2 x 2 contingency table of concordance. RESULTS: On the basis of pure observation alone, agreement among clinical tests on the presence and direction of lateral shift was 77% (kappa = 0.2; P < 0.248) and 79% (kappa = 0.4; P < 0.003), respectively. Agreement on the relevance of lateral shift and the lateral component according to symptom responses was 85% (kappa = 0.7; P < 0.000) and 92% (kappa= 0.4; P < 0.021), respectively. Using the repeated movements and static end-range loading strategy to define the centralization phenomenon and directional preference, agreement was 95% (kappa = 0.7; P < 0.002) and 90% (kappa = 0.9; P < 0.000), respectively. When patients with low back pain were classified into the McKenzie main syndromes and into specific subgroups, agreement was 95% (kappa = 0.6; P < 0.000) and 74% (kappa = 0.7; P < 0.000), respectively. CONCLUSIONS: Interexaminer reliability of the McKenzie lumbar spine assessment in performing clinical tests and classifying patients with low back pain into syndromes were good and statistically significant when the examiners had been trained in the McKenzie method. PMID- 11935121 TI - Development and validation of a new technique for assessing lumbar spine motion. AB - STUDY DESIGN: Dynamic lumbar flexion-extension motions were assessed by an electrogoniometer and a videofluoroscopy unit simultaneously. OBJECTIVE: To develop and validate a new technique for the assessment of lumbar spine motion. SUMMARY OF BACKGROUND DATA: Spine instability, a clinical condition that is common but difficult to diagnose, has been suggested to involve a characteristic change in the relation between vertebrae during motion. Assessment of lumbar instability using functional radiographs is controversial. Information regarding dynamic spine kinematics in vivo is limited. METHODS: A lumbar spine motion analysis system was developed, and its reliability was assessed. Simultaneous total flexion range of motion and segmental motion of the lumbar spine were assessed in 30 healthy volunteers. Lumbar images were captured in 10 degrees intervals during flexion-extension. Intervertebral flexion-extension of each vertebral level was calculated. RESULTS: In flexion, the lumbar vertebrae flexed with a descending order from L1 to L5 throughout the motion. Conversely, the concavity of lumbar lordosis increased steadily in extension. No statistically significant difference in the pattern of motion was found between genders. CONCLUSIONS: The results from this study showed that the newly developed technique is reliable. It may have potential value for evaluating spine instability in clinical practice. PMID- 11935123 TI - Treatment of craniocervical spine lesion with osteogenesis imperfecta: a case report. AB - STUDY DESIGN: A case report of craniocervical spine lesions including basilar impression, atlantoaxial dislocation, and syringomyelia, with osteogenesis imperfecta is presented, and the literature is reviewed. OBJECTIVE: To discuss the problems involved in the surgical management of craniocervical spine lesion with osteogenesis imperfecta. SUMMARY OF BACKGROUND DATA: Osteogenesis imperfecta is known to have various spine lesions as complications. However, few reports have described craniocervical lesions associated with osteogenesis imperfecta. METHODS: A 14-year-old girl with osteogenesis imperfecta, Silence classification IVB, experienced difficulty walking, with marked motor disturbance and muscle weakness in the extremities. Deep tendon reflexes were exaggerated bilaterally in the upper and lower extremities, and positive Babinski reflex and ankle clonus were observed bilaterally. Basilar impression, atlantoaxial dislocation, and syringomyelia were shown by plain radiography, tomography, three-dimensional computed tomography, and magnetic resonance imaging. RESULTS: In the reported patient, posterior fossa decompression and atlantoaxial posterior fusion could not be performed because the foramen magnum and upper cervical spine invaginated to the base of the skull. Therefore, occipitocervical spine fusion using titanium loop and wires was performed at the reduced position of the atlantoaxial dislocation, resulting in improvement of neurologic deficits. CONCLUSIONS: For patients with atlantoaxial dislocation, syringomyelia, and basilar impression without clinical symptoms or signs of brain stem compression, occipitocervical spine fusion alone at the reduction of the atlantoaxial dislocation may be indicated because these procedures improve neurologic deficits and prevent postoperative development of basilar impression and enlargement of syringomyelia. PMID- 11935122 TI - Incidental neoplasm in iliac crest autograft procured for anterior cervical fusion. AB - STUDY DESIGN: A case report is presented. OBJECTIVE: To present a previously unreported complication of procuring anterior iliac crest autograft. SUMMARY OF BACKGROUND DATA: Many complications that involve the common procedure of procuring iliac crest autograft have been reported and studied. The incidental finding of a neoplasm in an iliac crest autograft in an otherwise asymptomatic patient has not been reported previously. METHODS: A case of benign fibrous histiocytoma is presented as it was found incidentally during the procurement of the anterior iliac crest for anterior cervical fusion. Intraoperative decision making and alternatives for this challenging situation are reviewed. RESULTS: An iliac crest bone graft with a grossly unusual appearance was noted at the time of autograft procurement. The autograft was aborted, and an iliac crest allograft was substituted for the autograft to complete the planned cervical fusion. The lesion was later identified histopathologically and clinically as a benign fibrous histiocytoma. Postoperative computed tomography demonstrated the extent of the lesion. CONCLUSIONS: The intraoperative finding of a bony neoplasm at a planned autograft donor site in an otherwise healthy patient supports routine preoperative discussion of allograft alternatives. All patients undergoing surgery in which autograft is to be used should be informed regarding the possibility of allograft substitution. When a neoplasm is encountered unexpectedly, oncologic principles of obtaining frozen section or a touch-prep for diagnosis, avoiding the use of suspect graft material as well as cross contamination of iliac crest and anterior cervical surgical sites, should be applied. Care should be taken intraoperatively with potentially contaminated drapes and instruments. PMID- 11935124 TI - Do current results of endovascular abdominal aortic aneurysm repair justify more widespread use? PMID- 11935125 TI - A prospective study of sexual and urinary function before and after total mesorectal excision with autonomic nerve preservation for rectal cancer. AB - BACKGROUND: Oncologic resection of rectal cancer has been reported to be associated with a significant (10%-60%) rate of sexual and urinary dysfunction. We hypothesize that curative total mesorectal excision (TME) with autonomic nerve preservation (ANP) can be done with high rates of preservation of such function. STUDY DESIGN: We studied prospectively preoperative and postoperative urinary and sexual function in patients who had sphincter-preserving operations for rectal carcinoma without preoperative irradiation. Standardized questionnaires were used preoperatively and postoperatively, including the International Prostatic Symptom Score and a score of quality of urinary function satisfaction. Urodynamic evaluation was performed preoperatively and 3 months after the operation. The sexual results were evaluated after 1 year. RESULTS: Twenty patients, 13 men and 7 women, had TME, with ANP technique. Fourteen patients had coloanal anastomosis, 4 had a stapled colorectal anastomosis, and 2 had an ileoanal anastomosis. In all patients, hypogastric and sacral splanchnic nerves were identified and preserved. There was no mortality. Tumors are graded by Astler-Coller classification: A1 in 3 cases, A2 in 3, B1 in 7, B2 in 2, C2 in 1, and D in 1. There was no difference in preoperative and postoperative urinary function, International Prostatic Symptom Score, or urodynamic results, nor in the results of the quality of urinary function questionnaire. Four of the 7 women (69%) were sexually active before undergoing the surgical procedure. Sexual activity and ability to achieve orgasm was unchanged in these women. No dyspareunia was reported. Nine of the 13 men (69%) were sexually potent in the preoperative period. Sexual activity and potency were unchanged in these men. Retrograde ejaculation was reported in 1 man who previously had had normal antegrade ejaculation. After 3 months, 4 patients reported a reduced rigidity of erection, returning to normal by 1 year. CONCLUSIONS: The authors conclude that TME and ANP for cancer limited to the mesorectum do not impair urinary and sexual function. PMID- 11935126 TI - RET mutation profile and variable clinical manifestations in a family with multiple endocrine neoplasia type 2A and Hirschsprung's disease. AB - BACKGROUND: RET proto-oncogene germ line mutations are associated with the inherited multiple endocrine neoplasia type 2 syndromes (MEN 2), as well as with familial and sporadic Hirschsprung's disease (HSCR). In this study, we report a family in which the MEN 2A and the HSCR phenotypes are associated with a single point mutation in exon 10 of the RET proto-oncogene. Furthermore, we have investigated polymorphic sequence variants of the RET proto-oncogene. METHODS: Family members were tested for RET proto-oncogene mutations in exons 10, 11, 13, 14, 15, and 16 by double-gradient denaturing-gradient gel electrophoresis, nucleotide sequence analysis, and restriction endonuclease digestion of polymerase chain reaction products. The status of exon 2 and 13 polymorphic sites was investigated by EagI and TaqI digestion in 12 selected patients. RESULTS: A heterozygous C618R mutation of RET exon 10 was identified in 12 family members. Five out of 7 children with mildly elevated pentagastrin-stimulated calcitonin levels who carried the mutation underwent prophylactic thyroidectomy before the age of 12. C-cell hyperplasia (CCH) was found in 4 children and a microscopic medullary thyroid carcinoma (MTC) in an 8-year-old female. Neither CCH nor MTC was found in the only family member affected with HSCR, an 8-year-old male. This patient inherited the mutated RET allele from his mother, who had MTC but not HSCR, together with a rare allelic variant at codon 45 of RET exon 2. CONCLUSIONS: This report of a newly-described kindred with the infrequent clinical association between MEN 2A and HSCR confirms the risk of the latter phenotype among carriers of RET exon 10 cysteine codon mutations. Nevertheless, the influence of other genetic or environmental factors cannot be excluded. PMID- 11935127 TI - Invited commentary: Are specific inherited mutations important in clinical management of hereditary cancer syndromes? PMID- 11935128 TI - Nitric oxide synthase 2 mRNA expression in relation to p53 and adenomatous polyposis coli mutations in primary colorectal adenocarcinomas. AB - BACKGROUND: The inducible nitric (NO) synthase 2 (NOS2) is upregulated in breast, brain, colon, and gynecological tumors, which indicate that NO may have a role in tumorigenesis. However, little is known about the role and regulation of NOS2 in colorectal carcinomas. Recent in vitro experiments have implicated that NOS2 is downregulated by p53 accumulation. Virtual analysis of the NOS2 promoter showed putative TCF-4/Lef-1 response elements, which indicate a potential regulation of NOS2 expression by activation of the adenomatous polyposis coli (APC)/beta catenin pathway. METHODS: NOS2 mRNA expression was investigated in 59 colorectal carcinomas by reverse transcriptase/real-time polymerase chain reaction and related to mutations in the p53, APC, and beta-catenin genes. Presence of NOS2 protein was studied by Western blot, and the localization was studied by immunohistochemistry. Loss of heterozygosity was studied in the region of the NOS2 gene. RESULTS: The NOS2 mRNA and protein expression were significantly higher in tumors than in control tissue. Immunohistochemistry revealed extensive NOS2 staining in the epithelial cells and, to a minor degree, in leukocytes. Increased NOS2 mRNA expression was found in Dukes' stages A and B compared with the C and D stages. No relationship was found between elevated NOS2 expression and loss of heterozygosity in the later stages according to Dukes' classification or mutations in the p53, APC, or beta-catenin genes. CONCLUSIONS: Inactivating mutations in the p53 and APC pathways are not the main explanation for the increased NOS2 expression found in colorectal tumors. PMID- 11935129 TI - Platelet-derived growth factor-AA-mediated functional angiogenesis in the rat epigastric island flap after genetic modification of fibroblasts is ischemia dependent. AB - BACKGROUND: The aim of this study was to induce therapeutic angiogenesis in ischemically challenged flap tissue by means of gene transfer. METHODS: Isogenic rat fibroblasts were retrovirally transfected to produce platelet-derived growth factor (PDGF)-AA. Stable gene expression was monitored by PDGF-AA enzyme-linked immunosorbent assay. Eighty animals were divided into 2 groups (1 and 2), each with 4 subgroups. The angiogenic target was a 7 x 7-cm epigastric island flap used as a necrosis model. Group 1 received flap treatment 1 week before flap elevation: 10(7) genetically modified fibroblasts, expressing PDGF-AA (genetically modified fibroblasts) plus 1 mL of Dulbecco's modified Eagle's medium (DMEM) (1A), 10(7) nonmodified fibroblasts (NMFB) plus 1 mL of DMEM (1B), 1 mL of DMEM (1C), and 1 mL of sodium chloride 0.9% (1D). All substances were injected at evenly distributed spots into the panniculus carnosus of the entire flap. Group 2 had the same flap treatment at the day of flap elevation. All flaps were sutured back. Seven days later, the flaps were harvested and examined both clinically, histologically, and immunohistochemically. RESULTS: In vitro, the GMFB produced up to 117.9 +/- 57.2 ng of PDGF-AA/mL medium during a 4-day period, compared with 0.7 +/- 0.6 ng of PDGF-AA/mL medium produced by NMFB in the same time period. In vivo production of PDGF-AA in flaps amounted to 1.3 +/- 0.7 ng of PDGF-AA/1 microL flap tissue for group 1A and 1.7 +/- 1.1 ng of PDGF-AA/1 microL flap tissue for group 2A seven days after cell transplantation. Fibroblasts persisted in all flaps from groups 1A, 1B, 2A, and 2B without major inflammatory reaction. Clinically, group 2A developed significantly less flap necrosis compared with all other groups, including group 1A. Accordingly, only group 2A gave significant histologic and immunohistochemical evidence for enhanced angiogenesis within the flap tissue. CONCLUSIONS: After retroviral gene transfer, isogenic rat fibroblasts produce high amounts of PDGF-AA in vitro. In vivo, PDGF AA can be detected in flaps receiving genetically modified fibroblasts, which suggests survival of the implanted fibroblasts in this model. PDGF-AA produced by GMFB can induce flap angiogenesis only under ischemic conditions in this model. Transplantation of PDGF-AA-overexpressing fibroblasts results in higher flap survival in this model. PMID- 11935130 TI - A prospective randomized study comparing D2 total gastrectomy versus D2 total gastrectomy plus splenectomy in 187 patients with gastric carcinoma. AB - BACKGROUND: Classic surgical treatment of upper third gastric carcinoma is based on an extended total gastrectomy, including splenectomy. The purpose of this study was to perform a prospective randomized clinical trial comparing the early and late results of total gastrectomy (TG) versus total gastrectomy plus splenectomy (TGS). METHODS: One hundred eighty-seven patients with gastric carcinoma were included. In all patients a D2 total gastrectomy was performed. During surgery they were randomized to 1 of 2 operative options. They were monitored to their death or to 5 years later if they were alive. RESULTS: Operative mortality was similar after both operations (3% after TG and 4% after TGS). Septic complications after surgery were higher after TGS compared with TG (P <.04). Five-year survival rates were not statistically different between groups or in subset analysis according to stage of disease. CONCLUSIONS: On the basis of the results of the present prospective randomized trial, splenectomy is not necessary in early stages of disease. A low operative mortality rate (less than 3%) must be achieved to obtain good long-term results. PMID- 11935131 TI - Hemorrhage associated with vitamin C deficiency in surgical patients. AB - BACKGROUND: Diffuse hemorrhage in surgical patients with normal coagulation parameters may be caused by vitamin C deficiency and is rapidly reversed by vitamin C replacement. METHODS: Patients treated on a surgical service were entered into a clinical registry over a 12-month period if they experienced diffuse hemorrhage in the face of normal coagulation parameters and a plasma ascorbic acid level < 0.6 mg/dL (normal 0.6-2.0 mg/dL). Oral vitamin C replacement was administered after determination of plasma ascorbic acid level. Response to therapy, including subsequent bleeding events, need for blood transfusions, and demographic data including social and dietary history were retrospectively reviewed from hospital and outpatient clinic records. RESULTS: Twelve patients with bleeding diatheses and low plasma ascorbic acid levels were identified. Plasma ascorbic acid levels were 0.1 to 0.5 mg/dL (mean, 0.3 mg/dL). There were 6 men and 6 women; age ranged from 46 to 90 years (mean, 78 years). Coagulation parameters were normal in all patients. Diffuse postoperative bleeding from nonsurgical causes was evident in 10 of 12 patients. Four patients, 2 of whom had operations, presented with chronic recurrent blood loss from the gastrointestinal tract. Each patient received 250 to 1000 mg of vitamin C replacement daily. Within 24 hours of vitamin C administration, there was no further evidence of clinical bleeding nor need for subsequent blood transfusions in any patient. CONCLUSIONS: Vitamin C deficiency should be included in the differential diagnosis of nonspecific bleeding in surgical patients. Prolonged hospitalization, severe illness, and poor diet create vitamin C deficiency with significant clinical consequences. Oral vitamin C replacement rapidly reverses the effects of this disorder. PMID- 11935132 TI - Protective effects of ex vivo graft radiation and tacrolimus on syngeneic transplanted rat small bowel motility. AB - BACKGROUND: Intestinal transplantation is unduly complicated by the nontolerogenic properties of the gut-associated lymphoid tissue. Because simultaneous graft irradiation and bone marrow infusion significantly prolong the survival of the small bowel transplanted animal, our objective was to determine the functional motility effects of the immune modulating, graft irradiation procedure in the presence and absence of tacrolimus immunosuppression. METHODS: Four groups of syngeneic orthotopic small bowel transplanted animals were studied 48 hours after operations (untreated, tacrolimus, ex vivo graft irradiation, and tacrolimus + irradiation) and compared with controls. Histologic analysis was performed for mucosal apoptosis and neutrophilic infiltration into the muscularis externa. Gastrointestinal in vivo transit and in vitro circular muscle strip contractions were quantified in response to bethanechol (0.3-300 micromol/L). RESULTS: Graft irradiation ex vivo alone or in the presence of tacrolimus significantly increases (> 10-fold) the number of apoptotic mucosal cells after transplantation. Functional measurements showed that transplantation resulted in a significant delay in gastrointestinal transit and a decrease in muscle strip contractility. Tacrolimus and graft irradiation significantly ameliorated the transplant-induced dysfunction. CONCLUSIONS: Given the endowed propensity of mucosal regeneration, the immunologic and functional benefits of ex vivo graft irradiation appear to outweigh the detrimental effects to the mucosa. PMID- 11935133 TI - Soluble thrombomodulin plasma levels are an early indication of a lethal course in human acute pancreatitis. AB - BACKGROUND: The potential to predict severe disease and lethality by using plasma soluble thrombomodulin (sTM) and C-reactive protein (CRP) levels in 73 patients with acute pancreatitis was analyzed in a prospective 5-year investigation performed at a single institution. METHODS: According to the Atlanta criteria, pancreatitis was classified as mild in 23 patients and as severe in 50 patients. Blood was collected on days 1, 3, 5, 7, 10, 14, 21, and 28 after the onset of pain and analyzed for sTM and CRP. RESULTS: During the period between days 3 and 10 of acute pancreatitis when most of the admissions occurred, sTM levels at a cutoff of 75 ng/mL on day 3 (sensitivity, 100%; specificity, 77%; positive predictive value, 38%; negative predictive value, 100%) and 71 ng/mL on day 10 (sensitivity, 100%; specificity, 77%; positive predictive value, 41%; negative predictive value, 100%) were predictive of a lethal outcome. With sTM levels, it was not possible to differentiate patients with mild pancreatitis from those with severe pancreatitis (Atlanta classification). In contrast, CRP levels at a cutoff of 113 mg/L on day 3 differentiated severe from mild courses with a diagnostic sensitivity of 84%, a specificity of 60%, a positive predictive value of 78%, and a negative predictive value of 69%. CRP levels at a cutoff of 122 mg/L on day 10 differentiated mild from severe courses (nonsurvivors) with a diagnostic sensitivity of 72%, a specificity of 72%, a positive predictive value of 86%, and a negative predictive value of 53%. In contrast, differentiation of mild forms of acute pancreatitis from severe pancreatitis (survivors) on day 10 was not possible. CONCLUSIONS: CRP is a valuable marker of disease severity in acute pancreatitis especially in the first period of pancreatitis, whereas sTM identifies early those patients with the most severe courses and a high risk of dying (negative predictive value, 100%). Determination of sTM in addition to CRP offers the opportunity of identifying early those patients who require intensive care most urgently. Of course, further investigations of sTM in acute pancreatitis are indicated to confirm our results. PMID- 11935134 TI - Downstaging of hepatocellular carcinoma and liver metastases from colorectal cancer by selective intra-arterial chemotherapy. AB - BACKGROUND: Although resection is the sole chance of cure in patients with hepatocellular carcinoma (HCC) or metastatic colorectal cancer to the liver, most patients are not candidates for surgery at the time of diagnosis. Strategies aiming at downstaging large or multifocal tumors to enable curative resection are appealing. The aim of this study was to evaluate the effects of neoadjuvant selective intra-arterial chemotherapy in noncirrhotic patients with unresectable HCC or metastatic colorectal cancer to the liver in the absence of extrahepatic disease. METHODS: Selective chemotherapy was provided by using a subcutaneous pump device via a catheter placed in the gastroduodenal artery. Chemotherapy regimen included floxuridine (0.2 mg/kg/day for 14 days) in each patient with additional boluses of cisplatin and doxorubicin on day 1 of each cycle in the presence of HCC. Patients were evaluated at 3, 6, 9, and 12 months for possible curative resection. Complete follow-up was available for each patient. RESULTS: Twenty-eight patients with unresectable liver tumors (5 HCC and 23 metastatic colorectal cancer) were included in this study. There were no surgical complications related to pump insertion, and local chemotherapy was started within 1 week of surgery in each patient. The median follow-up in survivors was 31 months (range, 30 months to 5 years). Chemotherapy was well tolerated in 18 (64%) patients. Chemotherapy was discontinued in 4 patients because of abnormal liver function test results, and 2 of them required a biliary stent to relieve a biliary stricture. In 9 patients downstaging enabled curative resection (3 HCC, 6 colorectal metastasis). Seven of these patients were alive and tumor free at the completion of the study, with at least 2 years of follow-up. The actuarial survival rates at 3 years for HCC and colorectal metastases were 60% and 50%, respectively. CONCLUSIONS: About one third of patients with unresectable liver tumors can be successfully treated by neoadjuvant intra-arterial chemotherapy followed by curative resection. This strategy appears particularly promising in patients with large HCC. This approach should be investigated further. PMID- 11935135 TI - Frequency and predictive factors of malignancy in residual thyroid tissue and cervical lymph nodes after partial thyroidectomy for differentiated thyroid cancer. AB - BACKGROUND: The extent of surgery in differentiated thyroid cancer (DTC) has been a controversial issue. Total thyroidectomy potentially carries a higher operative risk, whereas partial thyroidectomy has the risk of leaving significant residual malignancy. The aim of this study was to assess the frequency and potential predictive factors of malignancy in the residual thyroid tissue and the cervical lymph nodes (CLN) in patients with DTC who had partial thyroid surgery and subsequently underwent completion thyroidectomy and/or modified neck dissection. Age, gender, pressure symptoms, duration of symptoms, size of the original tumor, tumor multifocality, perithyroidal tumor extension, soft tissue invasion, and serum thyroglobulin (Tg) level after first surgery were analyzed as potential predictive factors for the presence of malignancy in the thyroid remnant and the CLN. METHODS: We retrospectively reviewed the medical and pathologic data of 101 cases of DTC; 97 had papillary and 4 had follicular thyroid cancer. On the initial surgery, the median tumor size was 2.5 cm (range, 0.5 to 8.5 cm). Tumor multifocality occurred in 28 cases, perithyroidal tumor extension in 26 cases, and soft tissue invasion in 9 cases. Completion thyroidectomy was performed in 100 cases and modified neck dissection in 90 cases. RESULTS: On completion neck surgery, 39 patients had evidence of malignancy in the residual thyroid tissue and 36 patients in the CLN. In 23 (22.7%) cases, malignancy was present in both CLN and residual thyroid tissue. Only tumor multifocality and Tg level greater than 20 ng/mL after first surgery were predictive of the presence of malignancy in the thyroid remnant, whereas age older than 40 years, soft tissue invasion, perithyroidal tumor extension, and Tg level greater than 20 ng/mL were predictive of malignancy in CLN. CONCLUSIONS: Residual malignancy is common after partial thyroid surgery for DTC. Tumor multifocality and Tg level may be predictive of its presence in residual thyroid tissue. Age, perithyroidal tumor extension, soft tissue invasion, and Tg level are predictive of the presence of lymph node metastases. PMID- 11935136 TI - Effects of cortisol administration on hepatic circulation during brain death in rabbits. AB - BACKGROUND: Hemodynamic instability and endocrinologic alterations during brain death contribute to dysfunction of donor organs. The present study was conducted to examine the effect of cortisol administration on hepatic circulation in brain dead hypotensive donors. METHODS: Brain death was created by inflating a balloon catheter inserted into the epidural space in rabbits. In another set of experiments, non-brain-dead rabbits were bilaterally adrenalectomized. At 1 hour of equilibration after brain death or adrenalectomy, cortisol at a dose of either 0.05 or 5 mg/kg/hr was given intravenously for 5 hours. RESULTS: The induction of brain death resulted in a significant decrease in mean arterial pressure (MAP), whereas MAP after adrenalectomy was maintained. Both brain death and adrenalectomy caused an approximately 40% decrease in the portal vein blood flow (PVF) and hepatic tissue blood flow (HTF) and a significant increase in serum hyaluronic acid (HA) level as well as alanine aminotransferase (ALT) level. These changes were associated with a substantial decrease in serum cortisol level. Transmission electron microscopic examination showed swollen Kupffer cells with phagocytosis. Cortisol supplement after brain death or adrenalectomy significantly increased PVF and HTF to prevalues without affecting MAP. Both dosing rates of cortisol also significantly decreased serum HA and ALT levels and attenuated Kupffer cell activation. CONCLUSIONS: These results suggest that cortisol administration exhibits beneficial effects on hepatic circulation in hemodynamically unstable brain-dead animals. PMID- 11935137 TI - Cryoablation of unresectable pancreatic cancer. AB - BACKGROUND: Most patients with pancreatic cancer are not candidates for curative resection. The goal of this study was to evaluate the safety of an intraoperative ultrasound-guided cryosurgical procedure in a phase I study of unresectable pancreatic cancer. METHODS: From March 1995 to March 1999, 10 cryosurgeries using intraoperative ultrasound were performed on 9 patients with unresectable cancers at laparotomy. Four patients had a concurrent gastrojejunostomy, 2 had a chemical splanchnicectomy, and 1 underwent a concurrent hepatic cryosurgical procedure. RESULTS: There was no intraoperative morbidity or mortality. No patients developed postoperative pancreatitis or fistula. All patients had good pain control postoperatively and were tolerating a regular diet at the time of discharge. Pain control at discharge was achieved with an oral formulation (4/9), transdermal patch (3/9), no pain medication (1/9), and intravenous patient controlled analgesia (1/9). CONCLUSIONS: Ultrasound-guided cryoablation for unresectable pancreatic cancer appears safe and may contribute to improved postoperative pain control. Future studies to determine its therapeutic role in the management of unresectable pancreatic cancer are indicated. PMID- 11935138 TI - Small bowel obstruction due to intestinal migration of an inferior vena cava polytetrafluoroethylene graft. PMID- 11935139 TI - Progressive macrosyndactyly due to nerve territory-oriented lipofibromatosis: a case report. PMID- 11935141 TI - Inverse association between appendectomy and ulcerative colitis. PMID- 11935142 TI - Spontaneous intraperitoneal hemorrhage. PMID- 11935143 TI - Auto transplantation of the vermiform appendix. PMID- 11935144 TI - Biodiversity (Communications arising): suspect evidence of transgenic contamination. AB - Quist and Chapela claim that transgenic DNA constructs have been introgressed into a traditional maize variety in Mexico, and furthermore suggest that these constructs have been reassorted and introduced into different genomic backgrounds. However, we show here that their evidence for such introgression is based on the artefactual results of a flawed assay; in addition, the authors misinterpret a key reference to explain their results, concluding that reassortment of integrated transgenic DNA occurs during transformation or recombination. PMID- 11935145 TI - Biodiversity (Communications arising): maize transgene results in Mexico are artefacts. AB - Quist and Chapela's conclusion that the transgenes they claim to have detected in native maize in Oaxaca, Mexico, are predominantly reassorted and inserted into a "diversity of genomic contexts" seems to be based on an artefact arising from the inverse polymerase chain reaction (i-PCR) they used to amplify sequences flanking 35S transgenes from cauliflower mosaic virus (CaMV). PMID- 11935147 TI - Considerations for successful transplantation of encapsulated pancreatic islets. AB - Encapsulation of pancreatic islets allows for transplantion in the absence of immunosuppression. The technology is based on the principle that transplanted tissue is protected for the host immune system by an artificial membrane. Encapsulation offers a solution to the shortage of donors in clinical islet transplantation because it allows animal islets or insulin-producing cells engineered from stem cells to be used. During the past two decades three major approaches to encapsulation have been studied. These include intravascular macrocapsules, which are anastomosed to the vascular system as AV shunt; extravascular macrocapsules, which are mostly diffusion chambers transplanted at different sites; and extravascular microcapsules transplanted in the peritoneal cavity. The advantages and pitfalls of these three approaches are discussed and compared in the light of their applicability to clinical islet transplantation. All systems have been shown to be successful in preclinical studies but not all approaches meet the technical or physiological requirements for application in human beings. The extravascular approach has advantages over the intravascular because since it is associated with less complications such as thrombosis and infection. Microcapsules, due to their spatial characteristics, have a better diffusion capacity than macrocapsules. Recent progress in biocompatibility of microcapsules has brought this technology close to clinical application. Critical issues such as limitations in the functional performance and survival are being discussed. The latest results show that both issues can be solved by the transplantation of microencapsulated islets close to blood vessels in prevascularized solid supports. PMID- 11935148 TI - Uncoupling protein-2: evidence for its function as a metabolic regulator. AB - Uncoupling protein-2, discovered in 1997, belongs to a family of inner mitochondrial membrane proteins that, in general, function as carriers. The function(s) of uncoupling protein-2 have not yet been definitively described. However, mounting evidence suggests that uncoupling protein-2 could act in multiple tissues as a regulator of lipid metabolism. A role as a modulator of reactive oxygen species as a defence against infection is also postulated. In this review, a brief overview of the general and specific properties of uncoupling protein-2 is given and evidence for metabolic and immune regulatory functions is summarized. Uncoupling protein-2 could have particular importance in the regulation of lipid metabolism in adipose tissue and skeletal muscle. In addition, its ability to inhibit insulin secretion could also promote fat utilization over storage. Inhibition by uncoupling protein-2 of reactive oxygen species formation in macrophages and other tissues could have implications for regulation of immune function. The possibility of functions of uncoupling protein 2 in other tissues such as the brain are beginning to emerge. PMID- 11935149 TI - A phosphodiesterase inhibitor, cilostazol, prevents the onset of silent brain infarction in Japanese subjects with Type II diabetes. AB - AIMS/HYPOTHESIS: This study aimed to evaluate the effect of a phosphodiesterase inhibitor, cilostazol, on the prevention of silent brain infarction in diabetic patients without symptoms of vascular events. METHODS: A total of 89 subjects were allocated at random to the cilostazol group ( n = 43) or the control group ( n = 46). RESULTS: After the study period (3.2 +/- 0.5 years), carotid intima media thickness (IMT) (means +/- SD) had increased ( p < 0.01) by 0.18 +/- 0.19 mm in the control group. In the cilostazol group, intima-media thickness showed almost no change (-0.00 +/- 0.16 mm). In the control group, 2 out of 46 subjects showed symptomatic brain infarctions and 10 out of 34 subjects without infarct like region assessed by standard brain MRI examination showed silent brain infarctions after the observation period. On the other hand, no subjects in the cilostazol group showed silent brain infarction or strokes during the study period. Both at the beginning and end of the study period, the number of infarct like regions positively correlated with IMT ( r = 0.335, p < 0.001 or r = 0.347, p < 0.001 respectively). The progression of infarct-like regions was directly related to the increase in IMT during the study period ( r = 0.299, p = 0.004). CONCLUSION/INTERPRETATION: These data demonstrated that cilostazol could prevent the onset of silent brain infarction in Japanese subjects with Type II (non insulin-dependent) diabetes mellitus. Also, an increase in intima-media thickness of the carotid artery wall could be able to predict the onset of silent brain infarction. PMID- 11935150 TI - The pharmacokinetics, pharmacodynamics, safety and tolerability of NN2211, a new long-acting GLP-1 derivative, in healthy men. AB - AIMS/HYPOTHESIS: Glucagon-like peptide-1 (GLP-1), a polypeptide hormone secreted by the l-cells in the gastrointestinal tract, has shown promising effects as a new treatment modality for patients with Type II (non-insulin-dependent) diabetes mellitus. However, the pharmacokinetic profile of native GLP-1 with a rapid elimination has limited its therapeutic potential. NN2211 is a fatty acid derivative of GLP-1, which pre-clinically has shown a protracted pharmacokinetic profile, while maintaining its biological activity. This study aimed to investigate the safety, tolerability, pharmacokinetics and pharmacodynamics of NN2211 in healthy male subjects following seven days treatment. METHODS: In a double-blind, randomized, dose escalation, placebo controlled study, healthy male subjects were enrolled at five consecutive dose levels of NN2211 (1.25, 5.0, 7.5, 10.0, 12.5 microg/kg). Six subjects were allocated at random at each dose level to active or placebo treatment with a ratio of 2:1. Dosing with NN2211 was performed on day 1, and days 5-11. The 84-h pharmacokinetics and 24-h glucose and insulin profiles were assessed on day 1 and day 11. RESULTS: Following s. c. administration the half-life of NN2211 was found to be 12.6 +/- 1.1 h, with a subsequent accumulation index after a daily dose for seven days of 1.4-1.5. There were dose-proportional increases in exposure (AUC and C(max)) with increasing doses. Overall, there were no statistically significant differences from placebo in the 24-h glucose and insulin profiles. In subjects treated with NN2211 rather than placebo, there was a higher incidence of adverse events, most notably dizziness and adverse events related to the gastrointestinal system. There were no serious adverse events but three subjects were nonetheless withdrawn because of dizziness, fever and nausea. There were no clinically relevant changes in vital signs, ECG parameters, physical examination or safety laboratory parameters. A significantly lower diuresis was observed in the actively treated subjects, without a clinically relevant change in packed cell volume. CONCLUSIONS/INTERPRETATION: This study shows NN2211 has a pharmacokinetic profile supporting a daily dose in human beings, but also that subjects treated with NN2211 rather than placebo, had a higher incidence of adverse events, most notably dizziness and adverse events related to the gastrointestinal system. PMID- 11935151 TI - Angiotensin converting enzyme inhibiting therapy is associated with lower vitreous vascular endothelial growth factor concentrations in patients with proliferative diabetic retinopathy. AB - AIMS/HYPOTHESIS: Vascular endothelial growth factor (VEGF) is thought to be instrumental in the progression of diabetic retinopathy. Indications exist that the renin-angiotensin system is involved in VEGF overexpression. We assessed the vitreous VEGF concentrations in patients and related them to anti-hypertensive treatment, with special interest in the use of ACE-inhibitors. METHODS: Samples of vitreous fluid (10-80 microl) were obtained from 39 patients both with Type I (insulin-dependent) and Type II (non-insulin-dependent) diabetes mellitus and 11 non-diabetic patients undergoing intra-ocular surgery. The VEGF-A concentrations were assessed by immunoassay. RESULTS: Control patients and patients without proliferative diabetic retinopathy ( n = 8) had low and comparable VEGF concentrations (medians < 50 pg/ml). In contrast, patients with proliferative diabetic retinopathy ( n = 31) had high vitreous VEGF concentrations (median 1134 pg/ml), which showed a negative correlation with the use of ACE inhibiting medication (Spearman rank-R = - 0.54; p = 0.002, n = 13). Diastolic and systolic blood pressure did not differ significantly between the two subgroups with proliferative diabetic retinopathy, i. e. those patients receiving ACE-inhibition (medians 88/160 mm Hg, respectively) and the others (90/160). For the mostly used ACE-inhibitor in the proliferative diabetic retinopathy group, i. e. enalapril ( n = 8), a linear dose-effect relation was observed (-20 +/- 4 pg x ml(-1) x mg( 1) x day(-1); p = 0.024; coefficient +/- SEM). CONCLUSION/INTERPRETATION: Treatment with ACE-inhibitors attenuates retinal overexpression of VEGF-A in patients with proliferative diabetic retinopathy, probably by interference with a local effect of angiotensin II. PMID- 11935152 TI - Relation between glycaemic control, hyperinsulinaemia and plasma concentrations of soluble adhesion molecules in patients with impaired glucose tolerance or Type II diabetes. AB - AIMS/HYPOTHESIS: Increased plasma concentrations of circulating adhesion molecules in patients with Type II (non-insulin-dependent) diabetes mellitus could be associated with the increased cardiovascular risk in these patients. However, it is controversial whether increased adhesion molecule plasma concentrations are primarily related to hyperglycaemia or to hyperinsulinaemia. METHODS: We evaluated the plasma concentrations of soluble E-selectin, intracellular adhesion molecule-1 (ICAM-1), and vascular cell adhesion molecule-1 (VCAM-1) at baseline and during euglycaemic hyperinsulinaemic clamp in three different groups without additional cardiovascular risk factors: group A (control group), 28 healthy volunteers with normal glucose tolerance; group B, 24 subjects with fasting hyperinsulinaemia, normal fasting glucose but impaired glucose tolerance; group C, 32 patients with Type II diabetes, fasting hyperinsulinaemia and chronic hyperglycaemia. RESULTS: Plasma soluble E-selectin, ICAM-1, and VCAM 1 concentrations were higher ( p < 0.05) in patients with Type II diabetes (group C) compared with the other groups. The adhesion molecule concentrations correlate with the fasting plasma glucose ( r = 0.59, p < 0.001), the 2-h OGTT plasma glucose ( r = 0.70, p < 0.01), and the HbA(1 c) value ( r = 0.61, p < 0.05). The E-selectin but not the ICAM-1 and VCAM-1 plasma concentrations correlated with the fasting insulin concentrations ( r = 0.62, p < 0.05) or the whole body glucose uptake ( r = 0.59, p < 0.05) in the clamp. The hyperinsulinaemia during the euglycaemic hyperinsulinaemic clamp had no significant effect on the plasma concentrations of E-selectin, ICAM-1, and VCAM-1 in all three groups. CONCLUSION/INTERPRETATION: Our results suggest that increased E-selectin concentrations are related to hyperglycaemia, hyperinsulinaemia and insulin resistance, whereas increased ICAM-1 and VCAM-1 plasma concentrations in patients with Type II diabetes are rather related to hyperglycaemia than to hyperinsulinaemia or insulin resistance. PMID- 11935153 TI - Islet autoantibodies, nationality and gender: a multinational screening study in first-degree relatives of patients with Type I diabetes. AB - AIMS/HYPOTHESIS: First-degree relatives of patients with Type I (insulin dependent) diabetes mellitus diagnosed at 20 years of age or under were screened for islet cell antibodies (ICA) in the course of recruitment to an international diabetes prevention trial. Our aim was to evaluate the influence of age, gender, proband characteristics and nationality on the prevalence of ICA and co-existence of autoantibodies to GAD, IA-2 and insulin. METHODS: A central laboratory screened samples from 10 326 non-diabetic relatives who were aged less than 40 years, from eight European countries for ICA. Antibodies to GAD and IA-2 were measured in all samples with ICA of 10 JDF units or more. RESULTS: Overall, 8.9 % of relatives had ICA of 10 JDF units or more, 3.8 % with ICA of 20 JDF units or more. Of 921 relatives with ICA of 10 JDF units or more, 29 % had co-existing antibodies to GAD or IA-2 or both. ICA of 10 JDF units or more were more prevalent in males (10.8 %) than females (7.3 %). ICA with GAD or IA-2 antibodies or both were also more common in males (3.4 %) than females (1.9 %) and in relatives under 20 years of age (3.5 % vs 1.5 %). Multiple regression analysis showed nationality to be a determinant of ICA of 10 JDF units or more but not of ICA of 20 JDF units or more or of ICA with co-existing islet antibodies, and confirmed the importance of age and gender as determinants of islet autoimmunity. CONCLUSIONS/INTERPRETATION: Relatives from different European countries have similar rates of islet autoimmunity despite wide variation in the background incidence of childhood diabetes, and male excess is equally evident in all populations. The male excess of ICA and islet autoimmunity over 10 years of age reflects the higher male incidence of Type I diabetes in this age group, and suggests that boys may be more likely than girls to develop islet autoimmunity during adolescence. PMID- 11935154 TI - Combination therapy with low dose sirolimus and tacrolimus is synergistic in preventing spontaneous and recurrent autoimmune diabetes in non-obese diabetic mice. AB - AIMS/HYPOTHESIS: Sirolimus and tacrolimus are immunosuppressive drugs that prevent rejection of pancreatic islet allografts transplanted into patients with Type I (insulin-dependent) diabetes mellitus. This study aimed to determine whether sirolimus and tacrolimus can prevent autoimmune beta-cell destruction, and if so, what the mechanisms of action are. METHODS: Sirolimus and tacrolimus were given separately and together to female non-obese diabetic (NOD) mice from age 12 to 35 weeks. Diabetes incidence was determined and pancreatic insulitis and insulin content were measured. Sirolimus and tacrolimus were also given separately and together to diabetic NOD mice from the time of syngeneic islet transplantation until the reappearance of hyperglycaemia. Islet grafts were examined by RT-PCR assay for expression of interferon (IFN)- gamma, interleukin (IL)-2, IL-4, IL-10 and transforming growth factor (TGF)- beta1. RESULTS: Low doses of sirolimus (0.1 mg/kg) and tacrolimus (0.1 mg/kg) were synergistic in reducing insulitis, preserving pancreatic insulin content and preventing diabetes in female NOD mice (8 % diabetes incidence at 35 weeks vs 66 % in vehicle-treated mice). Also, the combination of sirolimus and tacrolimus prolonged syngeneic islet graft survival (median 34 days vs 13 days for vehicle-treated mice). Islet grafts from sirolimus plus tacrolimus-treated mice expressed significantly decreased mRNA contents of Th1-type cytokines (IFN- gamma and IL-2) and the highest ratio of TGF- beta1/IFN- gamma mRNA. CONCLUSION/INTERPRETATION: These findings suggest that combination therapy with sirolimus and tacrolimus prevent autoimmune beta-cell destruction by upregulating expression of the immunoregulatory cytokine, TGF- beta1 and reducing Th1 cytokines (IFN- gamma and IL-2) expressed in the islets. Low-dose sirolimus and tacrolimus combination therapy could warrant consideration for prevention or early treatment of human Type I diabetes. PMID- 11935156 TI - Metabotropic glutamate and GABA(B) receptors contribute to the modulation of glucose-stimulated insulin secretion in pancreatic beta cells. AB - AIMS/HYPOTHESIS: The neurotransmitters glutamate and gamma-aminobutyric acid (GABA) could participate in the regulation of the endocrine functions of islets of Langerhans. We investigated the role of the metabotropic glutamate (mGluRs) and GABA(B) (GABA(B)Rs) receptors in this process. METHODS: We studied the expression of mGluRs and GABA(B)Rs in rat and human islets of Langerhans and in pancreatic alpha-cell and beta-cell lines using RT-PCR and immunoblot analysis. Effects of mGluR and GABA(B) R agonists on insulin secretion were determined by radioimmunoassays and enzyme-linked immunoadsorbent assays (ELISAs). RESULTS: We detected mGluR3 and mGluR5 (but not mGluR1, 6 and 7) mRNAs in all of the samples examined. Trace amount of mGluR2 was found in MIN6 beta cells; mGluR4 was identified in rat islets; and mGluR8 expression was detected in rat islets, RINm5F and MIN6 cells. GABA(B)R1 a/b and 2 mRNAs were identified in islets of Langerhans and MIN6 cells. The expression of mGluR3, mGluR5, GABA(B)R1 a/b and GABA(B)R2 proteins was confirmed using specific antibodies. Group I (mGluR1/5) and group II (mGluR2/3) specific mGluR agonists increased the release of insulin in the presence of 3 to 10 mmol/l or 3 to 25 mmol/l glucose, respectively, whereas a group III (mGluR4/6-8) specific agonist inhibited insulin release at high (10-25 mmol/l) glucose concentrations. Baclofen, a GABA(B)R agonist, also inhibited the release of insulin but only in the presence of 25 mmol/l glucose. CONCLUSION/INTERPRETATION: These data suggest that mGluRs and GABA(B)Rs play a role in the regulation of the endocrine pancreas with mechanisms probably involving direct activation or inhibition of voltage dependent Ca(2+)-channels, cAMP generation and G-protein-mediated modulation of K(ATP) channels. PMID- 11935155 TI - Syntaxin-3 and syntaxin-1A inhibit L-type calcium channel activity, insulin biosynthesis and exocytosis in beta-cell lines. AB - AIMS/HYPOTHESIS: Syntaxin-1A (Syn-1A) is known to play a negative regulatory role in insulin secretion but the precise mechanisms for its action are not clear. Syn 2, -3 and -4 are also present in islet beta cells but their functions are not known. Here, we investigated the role of these syntaxins in the insulin secretory process. METHODS: We examined the following effects of Syn-1, -2, -3 and -4 expression in insulinoma beta-cell lines. Endogenous insulin secretion was measured by batch radioimmunoassay (RIA) and single cell patch clamp capacitance measurements. The L-type Ca(2+) channel activity was studied by patch clamp electrophysiology. Insulin gene transcription was examined by Northern blotting and measurement of insulin gene promoter activity by the co-expression of cyan fluorescent protein-labelled rat insulin promoter. RESULTS: Syn-1A or -3, but not Syn-2 or -4 overexpression, inhibited K(+)-induced insulin release as determined by RIA (49.7 +/- 5.5 % and 49.1 +/- 6.2 %, respectively) and electrophysiologic membrane capacitance measurements (68.0 +/- 21.0 % and 58.0 +/- 13.2 %, respectively). Overexpressed Syn-1A and -3, but not Syn-2, inhibited Ca(2+) channel current amplitude by 39.5 +/- 11.6 % and 52.7 +/- 6.0 %, respectively. Of note, overexpression of Syn-1A and -3 also reduced single cell (by confocal microscopy) and total cellular endogenous insulin content (by RIA) by 24.8 +/- 4.2 % and 31.8 +/- 3.9 %, respectively. This correlated to a reduction in endogenous insulin mRNA by 24.5 +/- 4.2 % and 25.7 +/- 4.2 %, respectively. This inhibition of insulin biosynthesis is mainly at the level of insulin gene transcription as demonstrated by an inhibition of insulin gene promoter activity (53.3 +/- 9.15 % and 39.0 +/- 6.8 %, respectively). CONCLUSIONS/INTERPRETATION: These results demonstrate that Syn-1A and -3 possess strong inhibitory actions on both insulin exocytosis and insulin biosynthesis whereas Syn-2 and -4 do not inhibit the insulin secretory process. PMID- 11935157 TI - Kidney function and glomerulopathy over 8 years in young patients with Type I (insulin-dependent) diabetes mellitus and microalbuminuria. AB - AIMS/HYPOTHESIS: We aimed to investigate prospectively the interrelation between kidney function and glomerular morphological changes over 8 years in young patients with Type I (insulin-dependent) diabetes mellitus and microalbuminuria. METHODS: Kidney biopsies were taken at baseline and after 8 years in 18 subjects who were 20 years of age (19-29 mean and range), had duration of diabetes for 11 years (7-18), and who had an albumin excretion rate of 45 microg/min (15-194). The glomerular ultrastructural parameters were analysed using stereological methods. RESULTS: At the end of the study three patients had an increased albumin excretion rate of more than 25 % a year, two of whom developed overt nephropathy. Glomerular filtration rate declined 2.3 ml/min x 1.73 m(-2) x yr(-1). Glomerular volume, volume fractions of matrix and mesangium, and basement membrane thickness showed an increase over the 8 years. Multiple regression analysis showed that mean 8-years HbA(1 c), matrix volume fraction(baseline) and basement membrane thickness BMT(baseline) accounted for 70 % of the variation in AER at the end of the study. Mesangial volume fraction(baseline,) glomerular filtration fraction(baseline,) and mean 8-year HbA(1 c) accounted for 73 % of the change in glomerular filtration rate from baseline. Smoking was strongly associated with the glomerular filtration rate at baseline ( r = 0.65). When glomerular filtration rate(baseline) was omitted from the equation, smoking was the only significant parameter linked to the change in glomerular filtration rate from the baseline. CONCLUSION/INTERPRETATION: In patients who had diabetes for 20 years, long-term hyperglycaemia and glomerulopathy found 8 years prior to the study, and possibly smoking, affected renal function (i. e. albumin excretion rate and glomerular filtration rate). PMID- 11935158 TI - Kinetics of integrin expression in the mouse model of proliferative retinopathy and success of secondary intervention with cyclic RGD peptides. AB - AIMS/HYPOTHESIS: Vitronectin-receptor-type integrins (alpha(v) beta(3) and alpha(v) beta(5)) are thought to be involved in the selective ablation of tumorigenic and other pathologic angiogenesis. Specifically, it has been shown that ligation inhibition of the alpha(v)-type integrins with cyclic penta-peptid peptide inhibits proliferative retinopathy by almost 80 % in a hypoxia-induced mouse model. METHODS: On the basis of growth factor and integrin expression dynamics in this model, secondary intervention approaches with cyclic RGDfV peptide were investigated. RESULTS: alpha(v)-integrin expression started immediately after induction of hypoxia (at postnatal day 12, p12) and persisted only during the initial period of neovascularization (until day p14). Vascular endothelial growth factor (VEGF) expression started at high values immediately after return of the mice into room air, and dropped rapidly to low values beyond day 13. In contrast, basic fibroblast growth factor (bFGF) was predominantly expressed during the phase of maximum angiogenesis which was noted between day p17 and 19. Based on these findings, cyclic penta peptide was administered subcutaneously at varying doses (2-20 microg/kg/day) for 5 days beginning either at day p14 (early intervention) or at day p17 (late intervention). Early secondary intervention showed a dose-dependent reduction of new vessels with maximum inhibition of 57 % (control 68.08 +/- 3.21 nuclei/section compared with RGDfV-treated 29.35 +/- 2.39 nuclei/section; p < 0.0001), whereas late secondary intervention had no effect. CONCLUSION/HYPOTHESIS: These data indicate that angiogenesis-related alpha(v)-integrin expression is VEGF- rather than bFGF dependent, and the efficacy of cyclic penta-peptid (RGDfV)-treatment in proliferative retinopathy is only effective as long as the alpha(v)-integrin target is prominently expressed. PMID- 11935159 TI - Decreased matrix degradation in diabetic nephropathy: effects of ACE inhibition on the expression and activities of matrix metalloproteinases. AB - AIMS/HYPOTHESIS: Extracellular matrix accumulation is thought to be involved in the pathogenesis of diabetic nephropathy. Increased matrix synthesis has been well documented but the effects of diabetes on degradative pathways, particularly in the in vivo setting, have not been fully explored. Furthermore, the effect of renoprotective therapies on matrix accumulation through these pathways has not been examined. We investigated the degradative pathway of type IV collagen and the effects of ACE inhibition in experimental diabetic nephropathy. METHODS: Diabetes was induced in 16 rats by administrating streptozocin; 8 of the diabetic rats were allocated at random to receive the ACE inhibitor perindopril (2 mg/l) in their drinking water and 8 age and weight matched rats served as controls. Gene expression of matrix metalloproteinase ( MMP) and tissue inhibitor of metalloproteinase ( TIMP) was measured by RT-PCR and type IV collagen content by immunohistochemistry. MMP activities were determined by degradation of a radiolabelled substrate and by zymography. RESULTS: Six months of diabetes was associated with a decrease in mRNA and enzymatic activity of MMP-9 (21 % and 51 % respectively, p < 0.05 vs control) and a 51 % increase in TIMP-1 mRNA ( p < 0.05 vs control). By contrast, MMP-2 mRNA was increased but its activity decreased (43 % and 43 % respectively, p < 0.05 vs control). Total degradative capacity of kidney tissue from diabetic rats was also lower (Control: 48 +/- 7 %, Diabetic: 33 +/- 6 %, p < 0.05). Activation of latent MMPs with amino-phenylmercuric acetate increased matrix degradation by two-fold. However the relative decrease associated with experimental diabetes still remained. All diabetes-associated changes in MMP and TIMP mRNA and activities were attenuated by perindopril treatment in association with reduced type IV collagen accumulation. CONCLUSIONS/INTERPRETATION: These results indicate that the impairment of matrix degradation contributes to matrix accumulation in diabetic nephropathy and that the beneficial effects of ACE inhibition could in part be mediated by modulation of changes in matrix degradative pathways. PMID- 11935160 TI - Polymorphism in the Calpain 10 gene influences glucose metabolism in human fat cells. AB - AIMS/HYPOTHESIS: A common G to A polymorphism ( UCSNP-43) in the Calpain 10 gene was recently found to be associated with Type II (non-insulin-dependent) diabetes mellitus and variations in post-absorptive and insulin stimulated glucose metabolism in vivo. We aimed to study the influence of Calpain 10 polymorphism on insulin action in fat cells. METHODS: Calpain 10 polymorphism ( UCSNP-19, -43 or 63) were set in relation to lipolysis and lipogenesis in isolated subcutaneous adipocytes of 46 apparently healthy non-obese subjects. RESULTS: For UCSNP-43 the G/G genotype had twofold higher basal and insulin stimulated rates as compared with AA/AG genotypes. However, there was no genotype effect on basal or insulin inhibited lipolysis rates in fat cells. The protein amount of GLUT 4 in adipocytes was not influenced by the polymorphism. Fat cells expressed mRNA for the Calpain 10 gene at a relatively high concentration, about 4 amol/microg RNA, which is similar to that of uncoupling protein-2. Neither a UCSNP-19 nor a UCSNP 63 polymorphism in the Calpain 10 gene was found to be associated with basal or insulin-induced adipocyte lipolysis and lipogenesis. None of the polymorphisms influenced body mass index or fasting plasma concentrations of insulin and glucose in 693 non-obese healthy subjects. CONCLUSIONS/INTERPRETATION: The Calpain 10 gene could be involved in the regulation of glucose metabolism but not lipolysis in human fat cells, although it does not involve adipocyte GLUT-4 protein content. It is possible that the Calpain 10 gene predisposes to diabetes by influencing the glucose metabolism. PMID- 11935161 TI - Adult-onset idiopathic Type I or ketosis-prone Type II diabetes: evidence to revisit diabetes classification. PMID- 11935162 TI - Liver transplantation and hepatitis C. AB - End-stage liver disease caused by chronic hepatitis C viral infection is one of the major indications for liver transplantation. However, evidence for ongoing viral replication can already be found days after surgery and may lead sequentially to lobular hepatitis, chronic active hepatitis, fibrosis and liver cirrhosis. In some patients, this evolution is remarkably fast, most probably enhanced by the immunosuppressive therapy. A minority of patients develop a clinical picture of progressive cholestatic liver disease with histological signs of chronic rejection, which may necessitate retransplantation. While the 1- and 5 year survival rates for all patients transplanted because of hepatitis C virus (HCV)-induced liver cirrhosis are satisfactory, severe complications of disease recurrence are nonetheless expected during the first and second decade after liver transplantation. Larger and preferably randomized studies are needed to investigate whether combination therapy with interferon and ribavirin, preferably initiated as soon as possible after liver transplantation, prevents the fast evolution to cirrhosis without the appearance of chronic rejection and the expected complications of recurrent end-stage HCV-induced liver disease. The final goal should be the inhibition of viral replication even before liver transplantation, but other antiviral strategies should probably be used to attain this goal in patients with decompensated cirrhosis. Although the recurrence of a hepatitis C infection and concomitant disease in the liver graft may cause substantial morbidity, end-stage liver disease and liver failure caused by a chronic hepatitis C infection remain good indications for liver transplantation. PMID- 11935164 TI - Adjunctive intracameral application of corticosteroids in patients with endothelial immune reactions after penetrating keratoplasty: a pilot study. AB - This study was performed to gain first experiences with a new adjunctive measure in the treatment of endothelial immune reactions after penetrating keratoplasty, i.e., intracameral injections of corticosteroids. In eight penetrating keratoplasty patients with mild endothelial immune reactions (IM1), 10 patients with moderate endothelial immune reactions (IM2) and 10 patients with severe endothelial immune reactions (IM3) intracameral injections of corticosteroids were performed within 24 h after referral to the clinic, following informed consent. All patients (IM1-3) received basic therapy of steroid eye drops and subconjunctival steroid injections; patients in group IM3 also received oral steroids. In IM1 patients eight of eight grafts, in IM2 eight of 10 grafts and in IM3 five of 10 grafts remained clear during a mean follow-up of 9.9 (1-20), 10.7 (3-17) and 9.6 (1-20) months after intraocular intervention. Only one of the 28 patients developed a further immune reaction during follow-up. No complications of intracameral injection were observed. Intracameral steroid injections thus seem to be a safe and helpful therapeutic measure in the treatment of moderate and severe endothelial immune reactions after penetrating keratoplasty. Further investigations in controlled randomized trials are necessary. PMID- 11935166 TI - Expression and inducibility of cytochrome P450 isoforms in 1-year-old intrasplenic liver cell transplants in rats. AB - Syngenic fetal liver tissue suspensions were transplanted into the spleens of 60- to 90-day-old male Fischer 344 inbred rats. Transplant recipients were compared with age-matched control rats. One year after surgery, the animals were treated orally with beta-naphthoflavone (BNF), phenobarbital (PB), dexamethasone (DEX) or the respective solvents 24 or 48 h before being killed. Expression of cytochrome P450 (P450) isoforms in spleens and orthotopic livers was assessed by immunohistochemistry and P450-dependent monooxygenase functions by the model reactions ethoxyresorufin O-deethylation (EROD), ethoxycoumarin O-deethylation (ECOD), pentoxyresorufin O-depentylation (PROD) and ethylmorphine N-demethylation (EMND). Spleens of control animals displayed almost no expression of P450 isoforms and P450-mediated monooxygenase functions. Similar to liver, in the transplanted hepatocytes no P450 1A1 but distinct P450 2B1 and 3A2 expression was observed. Furthermore, the transplant-containing spleens displayed significant EROD, ECOD, PROD and EMND activities. Similar to normal liver, BNF treatment enhanced P450 1A1 and 2B1, PB induced P450 2B1 and 3A2, and DEX induced P450 3A2 expression in the transplanted hepatocytes. Correspondingly, in the transplant containing spleens EROD, ECOD and PROD activities were significantly enhanced following BNF treatment, EROD, ECOD, PROD and EMND activities after PB administration, and EMND activity by DEX treatment. These results demonstrate that hepatocytes originating from fetal liver tissue suspensions can survive in the spleen at least for 1 year. They have differentiated into adult hepatocytes and even 1 year after transplantation express different P450 isoforms which are inducible by BNF, PB and DEX, corresponding to normal adult liver. PMID- 11935163 TI - Tacrolimus and low-dose steroid immunosuppression preserves bone mass after renal transplantation. AB - Bone loss, a recognized complication of renal transplantation (TP), is mainly attributed to steroids. The effect of other immunosuppressive agents on patients' bone mass is difficult to distinguish from that of steroids. In this study, we evaluate the evolution of bone mass density over the first 12 months following renal TP in two groups of patients given either low-dose steroids with tacrolimus ( n=7) or normal-dose steroids and cyclosporine ( n=19). Bone mineral density (BMD) of the lumbar spine, total hip, and hip subregions and total-body bone mineral content (BMC) were measured by dual-energy X-ray absorptiometry within the first 15 days, and 1 year after TP. Biological markers of bone turnover (serum calcium, phosphate, total alkaline phosphatase activity, intact parathyroid hormone, bone-specific alkaline phosphatase, calcitriol, and urinary pyridinolines) were regularly measured during follow-up. After TP, renal function improved rapidly in all patients. One year after TP, bone mass had decreased significantly in the cyclosporine group in all investigated sites. By contrast it had increased in the tacrolimus group. In order to compare the evolution of bone mass in patients given similar amounts of steroids, the cyclosporine group was subdivided in tertiles according to the 1-year cumulative oral intake of prednisolone. A significant bone loss was still observed in the low-steroid cyclosporine subgroup but not in the tacrolimus group, despite the similar steroids intake (3.5+/-0.5 g and 2.7+/-1 g, respectively). Bone gain in the tacrolimus group occurred despite a previous longer dialysis duration and a higher number of postmenopausal women who were not receiving hormone substitutes. Long-term evaluation of bone density (3-5 years post-TP) confirmed the bone gain in the tacrolimus patients. Interestingly, the profile of the biological markers of bone turnover appeared better in patients prescribed tacrolimus than in those given cyclosporine, though the differences did not reach statistical significance. Weconclude that tacrolimus associated with low-dose steroids might better preserve bone mass after renal TP than cyclosporine and normal doses of steroids. PMID- 11935165 TI - Combined effects of fasting and alanine on liver function recovery after cold ischemia. AB - The effect of donor nutritional status on hepatic function recovery after cold ischemia is still debated. We demonstrated previously that a 48-h fast diminished the survival rate of liver-transplanted rats and that the deleterious effect of fasting was prevented by infusion of alanine to the recipient at reperfusion. Whether the duration of fasting influenced the protective effect of alanine and whether this effect was metabolic were not known, and the elucidation of these questions is the aim of this study. The effect on hepatic function recovery of fasting periods of 24 h, 48 h and 72 h prior to cold ischemia were studied in a model of isolated, perfused rat liver. After a cold-ischemic time of 24 h in University of Wisconsin (UW) solution at 4 degrees C, livers were reperfused for 3 h. The combined effect of alanine (8 mM) infusion at liver reperfusion was evaluated for each prior fasting period. The addition of pyruvate (8 mM), a metabolic intermediary of alanine, was only tested in the 72-h fasting group. The evaluation criteria were: liver weight after reperfusion, release of aspartate aminotransferase (AST), alanine aminotransferase (ALT) and lactate dehydrogenase (LDH) in the perfusate, bile production, vascular resistance and liver histology after reperfusion. The enzyme release at reperfusion was significantly higher when livers were harvested from rats submitted to a 48-h fast (ALT) or a 72-h fast (ALT, AST, LDH), as compared to those from fed rats. Vascular resistance was increased in 72-h fasted livers. An addition of alanine (8 mM) at reperfusion lowered the release of AST, ALT and LDH. This effect was more obvious when the fasting duration was increased. By contrast, the addition of pyruvate at reperfusion did not improve the recovery of livers submitted to a 72-h fasting period before preservation. A long fasting period is deleterious as compared to feeding; however, this effect can be compensated by infusion of alanine at reperfusion. The mechanism involved is not metabolic. In a clinical setting, the infusion of alanine to the recipient at reperfusion may be a convenient way to compensate for donor undernutrition, especially after a long stay in an intensive care unit. PMID- 11935167 TI - Alloantigen-induced, T-cell-dependent production of nitric oxide by macrophages infiltrating skin allografts in mice. AB - The immunological rejection reaction occurring after organ or tissue transplantation is characterized by a strong infiltration of the graft by T cells and macrophages. Since the rejection reaction is highly specific, we tested the role of T cells in the activation of macrophages and in the induction of nitric oxide (NO) production during graft rejection. The rejection of both MHC and non MHC antigen-disparate skin allografts was associated with a significantly increased production of NO in the graft. The kinetics of NO production after transplantation correlated with the rejection reaction and with the fate of the allograft. A significant reduction in NO production was found in immunologically hyporeactive mice treated with cyclosporine, and no specific production of NO was found in tolerated skin allografts from neonatally tolerant mice. The production of NO was completely suppressed in graft explants from mice with depleted CD4(+) cells, but remained at a normal level in skin allografts from mice treated with anti-CD8 monoclonal antibody. The treatment of recipients of fully allogeneic skin grafts with 2-amino-5,6-dihydro-6-methyl-4H-1,3-thiazine (AMT), a specific inhibitor of the inducible NO synthase, resulted in a significant prolongation of graft survival. The results thus show CD4(+) T-cell-dependent, alloantigen induced production of NO by graft-infiltrating macrophages and the role of NO in the rejection reaction. We suggest that this pathway may represent one of the local effector mechanisms of graft rejection. PMID- 11935168 TI - Aplastic anemia after transplantation for non-A, non-B, non-C fulminant hepatic failure: case report and review of the literature. AB - Aplastic anemia is a rare complication of liver transplantation (<1%). However, an increasing number of cases of aplastic anemia have been recently reported when liver transplantation is performed for non-A, non-B, non-C fulminant hepatic failure. The aim of this study is to reevaluate the importance and the incidence of aplastic anemia after liver transplantation for non-A, non-B, non-C fulminant hepatic failure, and to propose preventive measures, diagnostic and management guidelines to try to reduce the incidence, morbidity and mortality associated with this complication. In this report a case of aplastic anemia after liver transplantation for non-A, non-B, non-C fulminant hepatic failure is described. In addition, the pertinent literature on aplastic anemia after liver transplantation, since the first description of that complication in 1987, is reviewed. A 20-year-old woman developed aplastic anemia 14 weeks after liver transplantation for fulminant non-A, non-B, non-C hepatitis. After failure of G CSF treatment, she was treated with intensive immunosuppression (FK 506, ATG, high-dose steroids). She is well 1 year post-transplantation, with normal liver tests and with bone marrow recovery. Through a Medline literature search (1988 1999), we identified 30 additional cases of aplastic anemia following liver transplantation for non-A, non-B, non-C fulminant hepatic failure. Of all liver transplantations performed for that indication at five participating centers, the mean incidence of aplastic anemia was 23.2%. Mean age was 10 years (1.2-29) and the male/female ratio was 4.6. For treating aplastic anemia, different modalities were used: ATG ( n=12), ALG ( n=1), OKT 3 ( n=1), G-CSF ( n=6), a 6-HLA compatible bone marrow transplantation ( n=3), and none ( n=12). The mortality rate remains high (39%), with infections and bleeding as the two most frequent causes of death. Based on this literature review, we conclude that aplastic anemia is a relatively common complication of liver transplantation for non-A, non-B, non-C fulminant hepatic failure in children and young adults. An unknown viral agent operating through immune-mediated mechanisms is probably responsible. The myelotoxic environment inherent to transplantation (e.g. azathioprine, trimethoprim) probably has a cumulative effect. Preventive measures (e.g. not using myelotoxic drugs) should be adopted in high-risk children and young adults transplanted for non-A, non-B, non-C fulminant hepatic failure. Early detection of bone marrow depression, a low threshold for performing a bone marrow biopsy, and prompt treatment are pivotal. Intensive standard supportive care with broad spectrum antibiotics and anti-fungal agents is essential during phases of pancytopenia. Although spontaneous recovery has been described under maintenance immunosuppression, increased immunosuppression, in particular with ATG, may reverse the aplastic anemia and promote bone marrow recovery. In unresponsive patients, six-HLA-identical bone marrow transplantation has been successful. PMID- 11935169 TI - A public forum to promote organ donation amongst Asians: the Scottish initiative. AB - There is a chronic shortage of organs for transplantation in the UK. This problem is particularly acute amongst Asians living within the UK. The Transplant Unit, University of Glasgow, joined forces with local businessmen to initiate a public meeting to promote awareness of transplant issues affecting Asians in the greater Glasgow area. During the Forum, we conducted a survey to determine the level of knowledge about organ transplantation, donation and willingness to donate, in relationship to the age, gender, marital status and religious affiliation amongst the attendees. The Forum was conducted at a public hall after publicity in the local press and Asian shops. The meeting was attended by over 300 people of Asian origin. Of the 90 survey forms handed out, 80 were returned fully completed. There was almost no opposition to organ donation, and many of the respondents were aware that religious leaders in the UK had endorsed organ donation. However, favourable disposition to these issues was not accompanied by carrying of the organ donor card, despite an awareness of the National Donor Register. The majority of the respondents were willing to undergo live organ donation, but were undecided about cadaveric donation. The issue of presumed consent drew mixed responses. Asians in the Glasgow region are not sympathetic to the matter of organ transplantation and donation, despite their recognition of the issues of organ shortage. We suggest that the matter needs to be further integrated into Asian culture by religious leaders and business persons. Our findings indicate that women over the age of 30 and based in the home may be in a unique position of influence by virtue of their position of centrality within the social network. This approach may also be suitable in other areas of the UK and the world with a large number of ethnic minorities. PMID- 11935170 TI - Alterations of glucocorticoid receptor expression during glucocorticoid hormone therapy in renal transplant patients. AB - Previous studies have demonstrated that glucocorticoid resistance develops in some patients during glucocorticoid therapy due to the down-regulation of glucocorticoid receptor (GCR) expression. A new flow cytometric method has been used for monitoring the intracellular GCR level in peripheral blood mononuclear cells (PBMCs). GCR expression by different lymphocyte subpopulations (bearing surface CD4+ CD8+ or CD19+) in steroid-treated (ST) and non-treated (NT) renal transplant patients was also compared. High, decreasing-dose steroid treatment caused a reduced GCR expression in 50% of the renal transplant patients. Long lasting, low-dose steroid therapy caused a fall in GCR level in only 11% of patients. CD4+ T lymphocytes showed both the lowest GCR level and its alteration due to steroid therapy, while CD8+ T cells and CD19+ B lymphocytes expressed higher GCR levels and were more sensitive to steroid treatment. Detection of GCR expression in lymphocytes may provide useful information about the changes of glucocorticoid sensitivity during the therapy. PMID- 11935171 TI - Splenohepatic arterial steal syndrome in liver transplantation: clinical features and management. AB - Well-known arterial complications after liver transplantation comprise thrombosis and major stenosis, which usually necessitate a retransplantation procedure. In our institution, in a series of 165 consecutive liver transplantations, we report the first recognized case of a splenohepatic arterial steal syndrome. This is characterized by an arterial malperfusion of the hepatic graft caused by a marked diversion of blood flow to a significantly enlarged spleen, which leads to major ischemic damage of the hepatic graft. After splenectomy the perfusion through the hepatic artery increased substantially and the graft was salvaged, with a following favorable clinical course. Splenohepatic arterial steal syndrome may ultimately result in graft loss if it is falsely diagnosed or recognized too late. A post-transplantation splenectomy represents a successful therapeutic approach; alternatively a primary arterial anastomosis to the aorta prevents the development of this condition. PMID- 11935172 TI - Short-term changes in cholesterol metabolism in 40 patients with liver transplants from living related donors. AB - After liver transplantation, there remains a need for precise markers for evaluation of grafts. We investigated whether serum cholesterol value can serve as a marker for evaluation of the transplanted liver during follow-up. The effect of liver transplantation involving living related donors was investigated in 40 recipients in terms of lipid metabolism as measured by serum cholesterol. The relationship between cholesterol value after transplantation and liver graft weight/body weight (LW/BW) was also examined. Serum cholesterol increased at 10 20 days post-transplantation in successful cases, stabilizing at a value of more than 100 mg/dl after 4 weeks post-transplantation. In unsuccessful cases, serum cholesterol showed little increase in the 3 weeks after transplantation, and thereafter continued to decline. Cholesterol levels never reached 100 mg/dl in any of the unsuccessful transplantation cases. It took 45 days on average for the serum cholesterol to reach 100 mg/dl in recipients with less than 1% LW/BW ratio graft, but only 10 days in recipients with more than 3% LW/BW ratio graft. Patients who had partial liver transplantation from living related donors showed rapid recovery of cholesterol synthesis. However, patients with liver grafts required an extensive period before normalization of cholesterol synthesis, suggesting a need for long-term follow-up of graft recipients. PMID- 11935173 TI - Compartment syndrome: an unusual complication of donor hepatectomy. PMID- 11935175 TI - Bacterial amino acid transport proteins: occurrence, functions, and significance for biotechnological applications. AB - Transport processes play a pivotal role in cellular metabolism, e.g. for the uptake of nutrients or the excretion of metabolic waste products. Moreover, they are also important in biotechnological processes such as the production of various amino acids by the use of microorganisms. The focus of this review is on bacterial amino acid transport systems, in particular those of Corynebacterium glutamicum and Escherichia coli, with respect to their function and biotechnological significance. PMID- 11935176 TI - Microbial production of vitamin B12. AB - One of the most alluring and fascinating molecules in the world of science and medicine is vitamin B12 (cobalamin), which was originally discovered as the anti pernicious anemia factor and whose enigmatic complex structure is matched only by the beguiling chemistry that it mediates. The biosynthesis of this essential nutrient is intricate, involved and, remarkably, confined to certain members of the prokaryotic world, seemingly never have to have made the eukaryotic transition. In humans, the vitamin is required in trace amounts (approximately 1 microg/day) to assist the actions of only two enzymes, methionine synthase and (R)-methylmalonyl-CoA mutase; yet commercially more than 10 t of B12 are produced each year from a number of bacterial species. The rich scientific history of vitamin B12 research, its biological functions and the pathways employed by bacteria for its de novo synthesis are described. Current strategies for the improvement of vitamin B12 production using modern biotechnological techniques are outlined. PMID- 11935177 TI - Importance of redox balance on the production of succinic acid by metabolically engineered Escherichia coli. AB - We had previously shown that succinic acid production in a pfl ldhA double mutant strain of Escherichia coli could be enhanced by amplifying the malic enzyme activity. However, recombinant E. coli NZN111 (F- Apfl::Cam ldhA::Kan) harboring pTrcML, a plasmid containing the E. coli malic enzyme gene, produced a considerable amount of malic acid along with the desired product, succinic acid. To have an insight into the intracellular metabolism, metabolic control analysis was carried out. From the results of a simulation, it was predicted that supplying additional reducing power could enhance succinic acid production. More reduced carbon substrate sorbitol was thus examined for the possibility of matching the potential during succinic acid production. When NZN111 (pTrcML) was cultured in LB medium containing 20 g sorbitol/l under a CO2 atmosphere, 10 g succinic acid/l was produced. The apparent yield of succinic acid was 1.1 g succinic acid/g sorbitol, which is 85% of the maximum theoretical yield. Therefore, it was found that redox balancing was important for the enhanced production of succinic acid in metabolically engineered E. coli. PMID- 11935179 TI - Purification of food-grade oligosaccharides using immobilised cells of Zymomonas mobilis. AB - Immobilised cells of the bacterium Zymomonas mobilis were used to remove glucose, fructose, and sucrose from food-grade oligosaccharide mixtures. Unpurified fructo , malto-, isomalto-, gentio-, and inulinoligosaccharides, containing total carbohydrate concentrations of 300 g l(-1), were added to immobilised cells, in 100 ml batch reactors. No pH control or nutrient additions were required. Contaminating glucose, fructose, and sucrose within the mixtures was completely fermented within 12 h. The fermentation end products were ethanol and carbon dioxide. A minor amount of sorbitol was also produced as a fermentation by product in the inulin-oligosaccharide mixture. No degradation of the oligosaccharides in the mixtures was observed. PMID- 11935178 TI - High-level ethanol production from starch by a flocculent Saccharomyces cerevisiae strain displaying cell-surface glucoamylase. AB - A Strain of host yeast YF207, which is a tryptophan auxotroph and shows strong flocculation ability, was obtained from SaccharomYces diastaticus ATCC60712 and S. cerevisiae W303-1B by tetrad analysis. The plasmid pGA11, which is a multicopy plasmid for cell-surface expression of the Rhyzopus oryzae glucoamylase/alpha agglutinin fusion protein, was then introduced into this flocculent yeast strain (YF207/pGA11). Yeast YF207/pGA11 grew rapidly under aerobic condition (dissolved oxygen 2.0 ppm), using soluble starch. The harvested cells were used for batch fermentation of soluble starch to ethanol under anaerobic condition and showed high ethanol production rates (0.71 g h(-1) l(-1)) without a time lag, because glucoamylase was immobilized on the yeast cell surface. During repeated utilization of cells for fermentation, YF207/pGA11 maintained high ethanol production rates over 300 h. Moreover, in fed-batch fermentation with YF207/pGA11 for approximately 120 h, the ethanol concentration reached up to 50 g l(-1). In conclusion, flocculent yeast cells displaying cell-surface glucoamylase are considered to be very effective for the direct fermentation of soluble starch to ethanol. PMID- 11935180 TI - Production of gamma-linolenic acid by Cunninghamella echinulata cultivated on glucose and orange peel. AB - A newly isolated strain of Cunninghamella echinulata grown on glucose produced significant quantities of biomass and cellular lipids in media with high C/N ratio. The oil yield from glucose consumed increased after nitrogen exhaustion in the growth medium, but gamma-linolenic acid (GLA) content in cellular oil systematically decreased during the lipid accumulation process. When lipid accumulation was completed, GLA concentration in the cellular lipids progressively increased. The highest GLA production (720 mg/l) was achieved in medium with a C/N ratio equal to 163. C. echinulata was also able to grow on orange peel. The C/N ratio in the orange peel decreased from 50 to 26 during solid-state fermentation. Maximum oxygen uptake was observed during assimilation of reducing sugars, whereas a polygalacturonase activity was detected after reducing sugars had been exhausted. The maximum GLA production was 1.2-1.5 mg/g of fermented peel, calculated on a dry weight basis. After enrichment of the pulp with inorganic nitrogen and glucose, an increase in the production of oil and GLA was observed. PMID- 11935181 TI - Single cell oil production by Yarrowia lipolytica growing on an industrial derivative of animal fat in batch cultures. AB - The growth of an oleaginous strain of Yarrowia lipolytica on an industrial fat composed of saturated free fatty acids (stearin) was studied. Lipid accumulation during primary anabolic growth was critically influenced by the medium pH and the incubation temperature. This process was independent of the nitrogen concentration in the culture medium, but was favored at a high carbon substrate level and at a low aeration rate. At pH 6 and a temperature of 28-33 degrees C, 9 12 g/l of dry biomass was produced, whereas significant quantities of lipids were accumulated inside the yeast cells (0.44-0.54 g of lipid per gram of biomass). The strain showed the tendency to degrade its storage lipids, although significant amounts of substrate fat, rich in stearic acid, remained unconsumed in the culture medium. Y. lipolytica presented a strong fatty acid specificity. The fatty acids C12:0, C14:0, and C16:0 were rapidly incorporated and mainly used for growth needs, while C18:0 was incorporated with reduced rates and was mainly accumulated as storage material. Reserve lipids, principally composed of triacylglycerols (55% w/w of total lipids) and free fatty acids (35% w/w), were rich in stearic acid (80% w/w), while negligible amounts of unsaturated fatty acids were detected. When industrial glycerol was used as co-substrate, together with stearin, unsaturated fatty acid concentration in the reserve lipid increased. PMID- 11935182 TI - Beta-galactosidase from a cold-adapted bacterium: purification, characterization and application for lactose hydrolysis. AB - The enzyme beta-galactosidase was purified from a cold-adapted organism isolated from Antarctica. The organism was identified as a psychotrophic Pseudoalteromonas sp. The enzyme was purified with high yields by a rapid purification scheme involving extraction in an aqueous two-phase system followed by hydrophobic interaction chromatography and ultrafiltration. The beta-galactosidase was optimally active at pH 9 and at 26 degrees C when assayed with o-nitrophenyl-beta D-galactopyranoside as substrate for 2 min. The enzyme activity was highly sensitive to temperature above 30 degrees C and was undetectable at 40 degrees C. The cations Na+, K+, Mg2+ and Mn2+ activated the enzyme while Ca2+, Hg2+, Cu2+ and Zn2+ inhibited activity. The shelf life of the pure enzyme at 4 degrees C was significantly enhanced in the presence of 0.1% (w/v) polyethyleneimine. The pure beta-galactosidase was also evaluated for lactose hydrolysis. More than 50% lactose hydrolysis was achieved in 8 h in buffer at an enzyme concentration of 1 U/ml, and was increased to 70% in the presence of 0.1% (w/v) polyethyleneimine. The extent of lactose hydrolysis was 40-50% in milk. The enzyme could be immobilized to Sepharose via different chemistries with 60-70% retention of activity. The immobilized enzyme was more stable and its ability to hydrolyze lactose was similar to that of the soluble enzyme. PMID- 11935183 TI - Utilization of ATP-binding cassette exporter for hyperproduction of an exoprotein: construction of lipase-hyperproducing recombinant strains of Serratia marcescens. AB - The Serratia marcescens extracellular lipase (LipA) is an enzyme applicable to enantioselective hydrolysis of racemic substrates. The enzyme is secreted through an ATP-binding cassette (ABC) exporter, the Lip system, encoded by the lipBCD genes. The S. marcescens recombinant carrying pLIPE121, which encodes the lipA gene in pUC19, exhibited a higher LipA production level than the wild-type strain. However, the level was lower than expected, and secretion was suggested to be a bottleneck. lipBCD plasmids were introduced into S. marcescens recombinants harboring lipA plasmids and the effectiveness of the lipBCD plasmids in elevating LipA productivity was investigated. S. marcescens strains harboring both lipA and lipBCD plasmids showed sevenfold greater extracellular LipA activity than the strain harboring the lipA plasmid alone. A high level of extracellular LipA production (1,300 kU/ml) and high plasmid stability (enough to carry out large-scale cultivation) were observed under non-selective conditions. Addition of L-proline and Tween 80 was effective in increasing cell growth of the recombinant, which led to high LipA production. In batch cultivation using a 30-l jar fermentor, LipA production was achieved at a high level of 5,200 kU/ml. This is the first report describing utilization of ABC exporter for the overproduction of an industrially important extracellular protein. PMID- 11935184 TI - Role of the general stress response during strong overexpression of a heterologous gene in Escherichia coli. AB - The strong overexpression of heterologous genes in Escherichia coli often leads to inhibition of cell growth, ribosome destruction, loss of culturability, and induction of stress responses, such as a heat shock-like response. Here we demonstrate that the general stress response, which is connected to the stress response regulator sigmas (sigma38, rpoS gene product), is suppressed during strong overproduction of a heterologous alpha-glucosidase. The mRNA levels of the rpoS and osmY stress genes drastically decrease after induction of the strong overexpression system. It is shown that an rpoS mutation causes a significant loss of cell viability after induction of the expression system. Furthermore, it is demonstrated that an E. coli c/pP mutant, which could be suggested to improve heterologous protein production, is not a good production host if a tac-promoter is used to control the expression of the recombinant gene. Data from this study suggest that the overexpression of the alpha-glucosidase was greatly decreased by sigma factor competition in the clpP mutant, due to the increased sigmas level in this mutant background. PMID- 11935185 TI - The level of pyruvate-formate lyase controls the shift from homolactic to mixed acid product formation in Lactococcus lactis. AB - Regulation of pyruvate-formate lyase (PFL) activity in vivo plays a central role in the shift from homolactic to mixed-acid product formation observed during the growth of Lactococcus lactis on glucose and galactose, respectively. Characterisation of L. lactis MG1363 in anaerobic batch cultures showed that the specific in vivo activity (flux) of PFL was 4-fold higher in L. lactis cells grown with galactose, compared with cells grown with glucose. The change in the PFL flux correlated with the observed variation in the PFL enzyme level, i.e. the PFL enzyme level was 3.4-fold higher in L. lactis cells grown on galactose than in those grown on glucose. To investigate whether a variation in the level of PFL was responsible for the shift in pyruvate metabolism, L. lactis strains with altered expression of pfl were constructed. The pfl gene was expressed under the control of different constitutive promoters in L. lactis MG1363 and in the PFL deficient strain CRM40. Strains with five different PFL levels were obtained. Variation in the PFL level markedly affected the resulting end-product formation in these strains. During growth on galactose, the flux towards mixed-acid products was to a great extent controlled by the PFL level. This demonstrates that a regulated PFL level plays a predominant role in the regulation of the metabolic shift from homolactic to mixed-acid product formation in L. lactis. PMID- 11935186 TI - Implication of manganese (III), oxalate, and oxygen in the degradation of nitroaromatic compounds by manganese peroxidase (MnP). AB - The fungal ligninolytic enzyme manganese peroxidase (MnP) is known to function by oxidizing Mn(II) to Mn(III), a powerful oxidant. In this work, an abiotic system consisting of Mn(III) in oxalate buffer under aerobic conditions (Mn(III)/oxalate/O2 system) was shown to be capable of extensively transforming 2 amino-4,6-dinitrotoluene (2A46DNT)--one of the main reduction products of 2,4,6 trinitrotoluene (TNT). No significant transformation occurred in the presence of other organic acids or under anaerobic conditions. The Mn(III)/oxalate/O2 system was also able to transform other nitroaromatic compounds such as 2-nitrotoluene, 4-nitrotoluene, 2,4-dinitrotoluene, TNT - the latter to a lesser extent -, and their reduction derivatives. The Mn(III)/oxalate/O2 system mineralized 14C-U-ring labeled 2A46DNT slightly, while no significant mineralization of 14C-U-ring labeled TNT was observed. Unidentified 14C-transformation products were highly polar. Electron spin resonance experiments performed on the Mn(III)/oxalate/O2 system revealed the generation of formyl free radicals (*COO-). The oxygen requirement for the transformation of nitroaromatic compounds suggests the involvement of superoxide free radicals (O2-*). produced through autoxidation of *COO- by molecular oxygen. The implication of such a Mn(III)/oxalate/O2 system in the MnP-catalyzed degradation of nitroaromatic pollutants by white-rot fungi is further discussed. PMID- 11935187 TI - Modeling growth and biochemical activities of Azospirillum spp. AB - An unstructured mathematical model was developed and used in the evaluation of biochemical activities of four Azospirillum spp. strains grown in batch cultures in a high C/N-ratio medium. The strains were evaluated for their ability to grow on fructose and produce exo-polysaccharide, and to sustain nitrogenase activity by using fructose or polysaccharides. Quantitative expression of the regulation of polysaccharide synthesis and nitrogenase (acetylene reduction) activity from the mineral nitrogen and sugar concentration in the culture medium was achieved. It was found that, during growth, Azospirillum spp. produced significant quantities of exocellular and capsular polysaccharide, whereas after depletion of the carbon source from the culture medium polysaccharides were consumed, especially in A. lipoferum strains. Significant nitrogenase activity was detected during polysaccharide degradation. Oxygen uptake was high during assimilation of fructose and low during polysaccharide degradation. PMID- 11935188 TI - Yield production, chemical composition, and functional properties of emulsifier H28 synthesized by Halomonas eurihalina strain H-28 in media containing various hydrocarbons. AB - Halomonas eurihalina strain H-28 is a moderately halophilic bacterium that produces an extracellular polysaccharide not only in media with glucose but also in media supplemented with hydrocarbons (n-tetradecane, n-hexadecane, n-octane, xylene, mineral light oil, mineral heavy oil, petrol, or crude oil). In this study we investigated yield production, chemical composition, viscosity, and emulsifying activity of exopolysaccharides (EPS) extracted from the different media used. The largest amounts of biopolymer were synthesized in media with glucose and n-hexadecane. Chemical composition varied with culture conditions; thus EPS from cultures grown in the presence of hydrocarbons had lower contents of carbohydrates and proteins than EPS from media with glucose. However, the percentages of uronic acids, acetyls, and sulfates were always higher than glucose EPS. Crude oil was the substrate most effectively emulsified. All EPS were capable of emulsifying crude oil more efficiently than the three control surfactants tested (Tween 20, Tween 80, and Triton X-100). All polymers gave low viscosity solutions. EPS H28 could be attractive for application in the oil industry and/or in bioremediation processes, bearing in mind not only its functional properties, but also the capacity of producer strain H-28 to grow in the presence of high salt concentrations and oil substrates. PMID- 11935189 TI - Degradation of biphenyl by Mycobacterium sp. strain PYR-1. AB - The metabolism of biphenyl by Mycobacterium sp. PYR-1 was investigated. The Mycobacterium sp. degraded >98% of the biphenyl added within 72 h. Analysis of ethyl acetate extracts of the culture medium by HPLC indicated that benzoic acid was the major metabolite. Other products were 4-hydroxybiphenyl, 4-hydroxybenzoic acid, and 5-oxo-5-phenylpentanoic acid. The metabolites were characterized by mass and 1H NMR spectrometry. Identification of benzoic acid and 5-oxo-5 phenylpentanoic acid indicates that biphenyl degradation by Mycobacterium sp. PYR 1 is generally similar to known pathways. A novel alternative metabolic pathway consisted of monooxygenation at C-4 of biphenyl to give 4-hydroxybiphenyl, with subsequent degradation via ring cleavage to 4-hydroxybenzoic acid. PMID- 11935190 TI - Analysis of alkali-soluble glucan produced by Saccharomyces cerevisiae wild-type and mutants. AB - The alkali-soluble glucan of the yeast cell wall contains beta-(1,3)- and (1,6)-D linkages and systemically enhances the immune system. To isolate Saccharomyces cerevisiae mutants producing glucan with a high degree of beta-(1,6)-D-glycosidic bonds, a wild-type strain was mutagenized with ultraviolet light. The mutants were then selected by treatment with 1.0 mg laminarinase, endo-beta-(1,3)-D glucanase/ml. The alkali-soluble glucan was extracted by modified alkalysis followed by the Cetavlon method and concanavalin-A chromatography. The prepared alkali-soluble glucans from the wild-type and the mutants were compared with respect to yield and polymer structure using gas chromatography, 13C-NMR spectrometry, high performance liquid, and multi-angle laser light scattering and refractive index detectors. The results indicated that the S. cerevisiae mutants had ten-fold more alkali-soluble glucan than the wild-type. Structural analysis revealed that the alkali-soluble glucan from the mutants also had a higher degree of beta-(1,6)-D-linkage than that from the wild-type. PMID- 11935191 TI - Responses of Mycobacterium sp. LB501T to the low bioavailability of solid anthracene. AB - Several recent reports have indicated that some bacteria may have adapted to the low bioavailability of hydrophobic environmental chemicals and that generalizations about the bioavailability of compounds such as polycyclic aromatic hydrocarbons (PAHs) may be inappropriate. Experimental evidence and theoretical considerations show that the utilization of PAHs requires bioavailability-enhancing mechanisms of the bacteria such as: (1) high-affinity uptake systems, (2) adhesion to the solid substrate, and (3) biosurfactant excretion. We examined possible specific physiological responses of anthracene degrading Mycobacterium sp. LB501T to poorly water-soluble anthracene in batch cultures, using solid anthracene as a sole carbon source. Mycobacterium sp. LB501T exhibited a high specific affinity for anthracene (a(o)A=32,500 l g(-1) protein h(-1)) and grew as a confluent biofilm on solid anthracene present as sole carbon source. No biofilm formation on anthracene was observed when excess glucose was provided as an additional substrate. This difference could be attributed to a modification of the cell surface of the bacterium. Anthracene grown cells were significantly more hydrophobic and more negatively charged than glucose-grown cells. In adhesion experiments, anthracene-grown cells adhered 1.5- to 8.0-fold better to hydrophobic Teflon and up to 70-fold better to anthracene surfaces than glucose-grown cells. However, no production of biosurfactants was observed. Our results thus indicate that attachment and biofilm formation may be a specific response of Mycobacterium sp. LB501T to optimize substrate bioavailability. PMID- 11935192 TI - Growth rate-dependent changes in Escherichia coli membrane structure and protein leakage. AB - The phospholipid and fatty acid content of the Escherichia coli membrane were investigated during continuous cultivation. At low growth rates, there was an increase in cardiolipin produced at the expense of phosphatidylethanolamine. Phosphatidylglycerol had a maximum at a growth rate of 0.3 h(-1). The amount of cyclic fatty acids was markedly increased at lower growth rates, while there was an evident minimum at 0.3 h(-1). This was also the case for saturated fatty acids. At this point, the unsaturated fatty acids had a maximum depending mainly on changes in cis-vaccenic acid. The mechanical strength towards sonication and osmotic shock/enzymatic treatment showed that the cells were more rigid at low dilution rates. However, this was accompanied by a higher cell lysis, a reduced capacity for total and specific protein production and a lower yield of cells. The amount of lipid A in the medium (endotoxin) was constant and negligible at all growth rates. The leakage of periplasmic protein to the medium had an optimum at 0.3 h(-1), resulting in a transport of 20% of the total recombinant product. It is argued that this constitutes the point of highest membrane fluidity and thus an increase possibility for protein transport. PMID- 11935194 TI - Inhibition of microbial H2S production in an oil reservoir model column by nitrate injection. AB - The effect of nitrate addition on microbial H2S production in a seawater-flooded oil reservoir model column with crude oil as carbon and energy source was investigated. Injection of 0.5 mM nitrate for 2.5-3.5 months led to complete elimination of H2S (initially 0.45-0.67 mM). The major decline in H2S level coincided with the first complete nitrate consumption and production of nitrite. When nitrate was excluded, H2S production resumed after approximately 2.5 months and reached previous levels after approximately 5 months. Using a fluorescent antibody technique, three populations each of sulfate-reducing bacteria (SRB) and nitrate-reducing bacteria (NRB) were monitored. SRB dominated the anoxic zone prior to nitrate addition, comprising 64-93% of the total bacterial population. The monitored NRB constituted less than 6% and no increase was observed during nitrate addition (indicating that other, unidentified, NRB populations were present). After 1-3 months without significant H2S production (3.5-5.5 months with nitrate), the SRB population collapsed, the fraction being reduced to 9-25%. The dominant SRB strain in the column, which constituted on average 94% of the monitored SRB population, was partly/completely inhibited by 50/75 microM nitrite in batch culture tests. Similar nitrite concentrations (50-150 microM) were detected in the column when the H2S level declined, indicating that nitrite inhibition was the main cause of H2S elimination. The results from this study indicate that nitrate/nitrite can be used to prevent detrimental SRB activity in oil reservoirs. PMID- 11935193 TI - Effects of randomly methylated-beta-cyclodextrins (RAMEB) on the bioavailability and aerobic biodegradation of polychlorinated biphenyls in three pristine soils spiked with a transformer oil. AB - The low bioavailability of polychlorinated biphenyls (PCBs) in soils often results in their slow and partial aerobic biodegradation. The process can be enhanced by supplementing soils with cyclodextrins. However, pure cyclodextrins are expensive and we have therefore explored the use of a less costly technical grade mixture of randomly methylated-beta-cyclodextrins (RAMEB). RAMEB was tested at 0, 1, 3 and 5% (w/w) in the aerobic bioremediation and detoxification of a loamy-, a humic- and a sandy-soil, each artificially contaminated with a PCB containing transformer oil (added PCBs: about 450 or 700 mg/kg), inoculated with an exogenous aerobic PCB-biodegrading bacterial co-culture and treated in slurry- and solid-phase laboratory conditions. Significant depletions of the spiked PCBs were observed in all microcosms of the three soils after 90 days of treatment; however, interesting yields of PCB dechlorination and detectable decreases of the original soil ecotoxicity were observed in the slurry-phase microcosms. RAMEB generally enhanced PCB-metabolism with effects which were dependent on the concentration at which it was applied, the physical-chemical nature of the amended soil, and the soil treatment conditions employed. RAMEB, which was slowly metabolized by soil microorganisms, enhanced the presence of PCBs and PCB cometabolizing bacteria in the soil-water phase, suggesting that RAMEB enhances aerobic biodegradation of PCBs by increasing pollutant bioavailability in soil microcosms. PMID- 11935195 TI - Natural attenuation potential of cyanide via microbial activity in mine tailings. AB - Biological removal by indigenous microflora of cyanide, contained in old (6-9 years) and fresh tailings (3 months), was studied in order to assess its natural attenuation potential via biodegradation. To investigate the presence of indigenous microflora in tailings, total heterotrophic and cyanide resistant bacteria were counted using the spread-plate method. The free cyanide mineralization potential was estimated using K14CN in the presence of various unlabeled cyanide concentrations (0, 5, and 10 mg CN/kg). The biodegradation of cyanide contained initially in the samples was also investigated by monitoring formate, formamide, ammonia and total cyanide (CNT) concentrations over 111 days. The enumeration of total heterotrophic and cyanide-resistant bacteria in old tailings showed an average population of 105 cfu/g. However, no growth was detected in fresh tailings. Nevertheless, cyanide mineralization tests indicated the presence, in both old and fresh tailings, of a cyanide-degrading microflora. In old tailings, maximum mineralization percentages of free cyanide ranging from 85% to 100% were obtained after 65 days at all concentrations tested. A mineralization percentage of 83% after 170 days was also observed in fresh tailings. No decrease of total cyanide concentration in old tailings was observed when the biodegradation of endogenous cyanide was tested whereas a significant decrease was recorded in fresh tailings after 96 days. The presence of strong metal-cyanide complexes resistant to biodegradation could explain the absence of biodegradation in old tailings. This study demonstrated the presence of an indigenous free cyanide-degrading microflora in both old and fresh tailings, and suggests that natural attenuation of cyanide in gold mine tailings is likely to occur via microbial activity. PMID- 11935196 TI - Bioremediation of toxic chromium from electroplating effluent by chromate reducing Pseudomonas aeruginosa A2Chr in two bioreactors. AB - The chromate-reducing ability of Pseudomonas aeruginosa A2Chr was compared in batch culture, with cells entrapped in a dialysis sac, and with cells immobilized in an agarose-alginate film in conjunction with a rotating biological contactor. In all three systems, the maximum Cr(VI) reduction occurred at 10 mg Cr(VI)/l. Whereas at 50 mg Cr(VI)/l concentration, only 16% of the total Cr(VI) was reduced, five spikings with 10 mg chromate/l at 2-h intervals led to 96% reduction of the total input of 50 mg Cr(VI)/l. Thus maximum Cr(VI) reduction was achieved by avoiding Cr(VI) toxicity to the cells by respiking with lower Cr(VI) concentrations. At 10 mg Cr(VI)/l, the pattern of chromate reduction in dialysis entrapped cells was almost similar to that of batch culture and 86% of the bacterially reduced chromium was retained inside the dialysis sac. In electroplating effluent containing 100 mg Cr(VI)/l, however, the amount of Cr(VI) reduced by the cells immobilized in agarose-alginate biofilm was twice and thrice the amount reduced by batch culture and cells entrapped in a dialysis sac, respectively. PMID- 11935197 TI - Magnetic resonance imaging of the hindfoot with surgical correlations. AB - The hindfoot is an anatomical region with a complex anatomy and it is very important from the biomechanical point of view. From the functional point of view the hindfoot may be divided into two different anatomical components: the talar foot and the calcaneal foot. Many bony and ligamentous structures, complexly linked together, form this anatomical region. This review aims to analyze the complex anatomy and the most frequent pathological conditions involving the hindfoot. The difficult surgical approach to this region makes imaging, in particular MRI, of fundamental importance for clinicians in order to give complete information about the bony, ligamentous or chondral structure involved. PMID- 11935198 TI - Sonographic findings of tensor fascia lata tendinopathy: another cause of anterior groin pain. AB - OBJECTIVE: To describe the sonographic appearances of the normal tensor fascia lata (TFL) origin and to describe the sonographic changes that were present in the TFL of a group of athletes presenting with anterior groin pain. DESIGN: The sonographic appearances of the TFLs of 40 healthy asymptomatic volunteers were evaluated. The TFLs of approximately 200 patients aged between 16 and 55 years presenting with anterior groin pain were evaluated in the course of routine sonographic assessment of the hip. Twelve abnormal TFLs were identified in 12 patients aged between 16 and 53 years. A retrospective review of the appearance of the normal and abnormal tendon with respect to its overall size and echotexture was made. RESULTS: The normal TFL has a thin ribbon-like appearance with a clearly defined fibrillar pattern and a mean anteroposterior (AP) size of 2.1 mm (range 1.5-3.1 mm). In the patient cohort the TFL typically appeared enlarged and contained a cone-shaped area of hypoechogenicity based on the iliac crest within the deep fibers of the origin. The TFL origin was enlarged up to 2 1/2 times its normal size. The mean AP size of the abnormal TFL was 4.7 mm (range 3.1-7.0 mm). CONCLUSION: Tendinopathy of the TFL is a cause of anterior groin pain. Sonography can be used to depict changes in the TFL, confirming the diagnosis and assessing the severity of the tendinopathy. PMID- 11935199 TI - Sonography of injury of the posterior cruciate ligament of the knee. AB - OBJECTIVE: To describe the sonographic appearance of injury of the posterior cruciate ligament (PCL) of the knee. DESIGN AND PATIENTS: Eleven patients (6 men, 5 women) with an average age of 36 years (range 17-70 years), referred for magnetic resonance (MR) imaging evaluation of knee pain, had PCL injury according to standard MR imaging criteria. Sonography was performed after the MR examination by the author, who was aware of the MR findings. Both the affected and contralateral asymptomatic PCL were scanned sonographically in the longitudinal plane with a curvilinear transducer, and their diameters were measured on the scanner using electronic calipers. Bilateral PCL sonography and measurement were also performed on 12 patients with other acute knee injuries but normal PCLs according to MR imaging ("non-PCL-injured knee cohort"), and on 10 normal volunteers. RESULTS: All PCLs were hypoechoic, regardless of injury. The injured PCLs had an average diameter of 0.88 cm (range 0.54-1.21 cm), while the contralateral PCLs had an average diameter of 0.51 cm (range 0.30-0.84 cm) (signed rank test, P<0.001). One patient also had focal discontinuity of the PCL. There was no statistical difference between the diameters of the right and left PCLs of the non-PCL-injured knee cohort (signed rank test, P=0.1) nor between those of the volunteers (signed rank test, P=0.6). The average difference in diameter between the injured and contralateral PCLs was statistically significant versus the difference between the non-PCL-injured knee cohort's PCLs (Mann Whitney test, P=0.0001) and versus the difference between the volunteers' PCLs (Mann-Whitney test, P=0.0001). CONCLUSION: PCL injury is manifested sonographically either as enlargement of the entire ligament, appreciated by comparison with the contralateral normal PCL, or by focal disruption of the ligament. PMID- 11935200 TI - Nodular fasciitis: correlation of MRI findings and histopathology. AB - OBJECTIVE: To compare the histopathology of nodular fasciitis (NF) with the magnetic resonance imaging (MRI) findings in order to evaluate the basis of the MR signal characteristics. DESIGN AND PATIENTS: Ten patients with NF, nine females and 1 male, with an age ranging from 13 to 58 years (mean 26.8 years) were studied. MRI findings, available in all 10 patients, were compared with the histopathology in nine patients, and an area-to-area comparative study of the whole specimen section histopathology and MRI was performed in two patients. RESULTS: On the basis of an excisional biopsy or resection specimen, the nine lesions were classified into myxoid ( n=4), cellular ( n=3) and fibrous ( n=2) subtypes. Four myxoid lesions with a subcutaneous location showed a homogeneous SI comparable with muscle on T1-weighted images, high SI on T2-weighted images, and had homogeneous enhancement. One cellular lesion presented with homogeneous, slightly higher SI than muscle on T1-weighted images and inhomogeneous, high SI on T2-weighted images. Alcian blue stain of the whole specimen section revealed the lesion had two parts corresponding to different enhancement patterns on MRI. The blue-stained myxoid part showed markedly diffuse enhancement, while the non stained cystic space had only peripheral enhancement. Two other cellular lesions had the same appearance on both T1- and T2-weighted images and showed inhomogeneous, diffuse enhancement. One fibrous subtype lesion presented with inhomogeneous, overall slightly higher SI than muscle on T1-weighted images, lower SI at the periphery and high SI in the center on STIR images and only peripheral enhancement. Microscopy and CD-31 staining of the lesion showed more extracellular matrix, with poor vascularity in the center and more collagenous matrix with higher vascularity at the periphery. CONCLUSION: Although similar findings were found in some lesions, the large histologic variability of NF hampers the definition of a prototype of NF on MRI. However, the MRI appearance of the myxoid subtype is rather characteristic. Histologic findings reflect the different SI characteristics and enhancement pattern on MRI. PMID- 11935201 TI - Percutaneous vertebroplasty in the treatment of vertebral body compression fracture secondary to osteogenesis imperfecta. AB - Percutaneous vertebroplasty, a minimally invasive interventional radiological procedure, has recently been used effectively for the treatment of symptomatic vertebral body compression fractures. Primary indications for vertebroplasty include osteoporotic compression fracture, osteolytic vertebral metastasis and myeloma, and vertebral hemangioma. We present a case and extend the indication of percutaneous vertebroplasty in a patient with a vertebral body compression fracture secondary to osteogenesis imperfecta. PMID- 11935202 TI - Skin metastasis from conventional giant cell tumor of bone: conceptual significance. AB - A conventional giant cell tumor of the proximal femur recurred twice locally and developed pulmonary nodules. The lung lesions were felt to be an example of "benign" metastases. Eight months after the initial presentation, the patient developed a single skin nodule on the contralateral leg. Histologic features of the skin nodule showed conventional giant cell tumor identical to the bone lesion. This nodule is a manifestation of arterial metastasis typical of any malignant tumor and seemingly contradicts the concept of "benign" metastasis. PMID- 11935203 TI - Intra-articular elastofibroma of the shoulder joint. AB - A 19-year-old man presented with an elastofibroma in his left shoulder joint. The patient had had limitation of motion in his left arm for 3 years, especially when rotating internally. Radiography of his left shoulder showed bone erosion in the neck of the humerus. On MR imaging, a soft tissue mass was noted in the shoulder joint eroding bone. The mass showed similar signal intensity to that of surrounding muscles on T1- and T2-weighted images. At surgery, a soft, encapsulated mass was found attached to the subscapularis muscle. It was pathologically confirmed as an elastofibroma. This unusual manifestation of an elastofibroma is discussed. PMID- 11935204 TI - Focal hematopoietic hyperplasia of the rib. AB - We report a case of focal hematopoietic hyperplasia in the rib of a 24-year-old woman. This is only the fourth case to be reported in the English literature, all of which have involved the rib. Radiologically they all manifested as an expansive and radiolucent lesion and contained ill-defined areas of increased density or calcification. Histologically, all have been characterized by mixed areas of hypercellular marrow and fatty marrow. The lesion is considered a form of pseudotumor. Treatment in our case was by wide marginal excision of the rib. PMID- 11935205 TI - Osteoblastoma-like osteosarcoma of the distal tibia. AB - We report a case of a 14-year-old boy with an intracompartmental lytic lesion with poorly defined margins in the right distal tibia that was originally treated with curettage and bone grafting. Histologic examination showed an osteoblastic tumor with unusual features, which was found on consultation to be an osteoblastoma-like osteosarcoma, a rare, low-grade variant of osteosarcoma. Subsequently, the patient underwent en bloc resection of the distal tibia, which was replaced with vascularized bone graft and followed by chemotherapy. Two years later, he is alive with lung metastases. PMID- 11935206 TI - Grebe syndrome with bilateral fibular hemimelia and thumb duplication. AB - Grebe syndrome is a rare recessively inherited form of short-limbed dwarfism. Among the skeletal anomalies reported in the past, complete fibular hemimelia and thumb duplication have not been documented. We report a case of Grebe syndrome with these associated anomalies and review the various skeletal anomalies reported in the literature related to this syndrome. Awareness of the skeletal anomalies that can occur in this syndrome should enable an accurate diagnosis. PMID- 11935209 TI - Dihydrotestosterone (DHT) modulates the ability of NSAIDs to induce apoptosis of prostate cancer cells. AB - PURPOSE: Recent evidence indicates that nonsteroidal antiinflammatory drugs (NSAIDs) are effective in the treatment and prevention of prostate cancer. In the study reported here, we investigated the ability of the steroid hormone dihydrotestosterone (DHT) to modulate NSAID-induced apoptosis of prostate cancer cells. MATERIALS AND METHODS: Using in vitro models of androgen-sensitive and androgen-insensitive human prostate cancer cells, we evaluated the ability of a specific cyclooxygenase-2 inhibitor (NS-398) and a nonspecific cyclooxygenase inhibitor (indomethacin) to induce apoptosis in the presence of various concentrations of DHT. Apoptosis was quantified using the TUNEL method and verified by electron microscopy. RESULTS: We found that increasing concentrations of DHT significantly enhanced the ability of NS-398 and indomethacin to induce apoptosis of androgen-sensitive LNCaP cells. The ability of NSAIDs to induce apoptosis of androgen-insensitive PC-3 cells, however, was not affected by the presence of DHT. Higher levels of DHT in the incubation medium both before as well as following exposure to NSAIDs enhanced apoptosis of LNCaP cells. Another steroid hormone that interacts with the androgen receptor in LNCaP cells (progesterone) also promoted apoptosis of these cells. Increasing concentrations of DHT caused LNCaP cells to shift from the S and G(2)/M to the G(0)/G(1) stages of the cell cycle. CONCLUSIONS: These observations support the use of DHT in combination with NSAIDs in the treatment of prostate cancer, and indicate that DHT is an important issue to address in clinical trials of NSAIDs since androgen ablation is a common treatment for prostate cancer. PMID- 11935210 TI - Triclosan inhibits enoyl-reductase of type I fatty acid synthase in vitro and is cytotoxic to MCF-7 and SKBr-3 breast cancer cells. AB - BACKGROUND AND PURPOSE: Human type I fatty acid synthase has been proposed as a chemotherapeutic target for the treatment of breast cancer based on the inactivation of human beta-ketoacyl synthase activity by cerulenin. Triclosan, a common antibiotic, functions by inhibiting the enoyl-reductase enzymes of type II fatty acid synthases in susceptible bacteria. If triclosan is an inhibitor of human fatty acid synthase and if inhibition of fatty acid synthase is toxic to breast cancer cell lines, triclosan could prove to be a lead compound for the treatment of breast cancer. Consequently, the inhibitory activity of triclosan against vertebrate type I fatty acid synthases and its effects on breast cancer lines in cell culture were investigated. METHODS: The inhibitory activities of triclosan against human and goose fatty acid synthases and each of the partial reactions were investigated using spectrophotometric assays. The ability of triclosan at various concentrations to inhibit growth and reduce the viability of MCF-7 and SKBr-3 cells in culture was evaluated. RESULTS: Kinetic studies showed triclosan to be a slow binding inhibitor of human and goose type I fatty acid synthase and to inhibit the partial activity of enoyl-reductase with IC(50) values between 10 and 50 microM. Triclosan at similar concentrations was also shown to inhibit both viability and growth of MCF-7 and SKBr-3 cells in culture. CONCLUSIONS: The results corroborate the hypothesis that fatty acid synthase may be a target of breast cancer chemotherapy and suggest that inhibitors of the enoyl-reductase partial activity of fatty acid synthase may have chemotherapeutic potential. PMID- 11935211 TI - Factors influencing the cellular accumulation of SN-38 and camptothecin. AB - PURPOSE: The influence of biophysical factors (drug metabolism, transport proteins, and chemical stability) on the cellular accumulation of camptothecin (CPT) and SN-38 was examined. METHODS: Drug transporter RNA transcript levels were measured by real-time reverse transcriptase polymerase chain reaction (RT PCR). Intracellular drug accumulation, metabolism, and drug stability studies were all performed by HPLC. RESULTS: A panel of three human cell lines exhibiting different drug resistant phenotypes was investigated. HT29 colon cells glucuronidated SN-38 but did not express P-gp or MRP1 or 2. HCT116 colon cells expressed P-gp and MRP2 but did not catalyse conjugation. A2780 ovarian cells neither catalysed drug metabolism nor contained these drug transporters. In all lines, SN-38 lactone was rapidly taken up achieving peak concentrations at the earliest time point studied (5 min, 3.3-4.1 ng/10(6) cells). Subsequently, a fall in intracellular lactone concentration occurred, stabilising after 4 h at 0.48 1.18 ng/10(6) cells. No significant differences in intracellular levels of lactone were observed between the three cell lines with one exception: a twofold increase in HCT116 cells at 24 h. Stability studies in culture medium revealed that SN-38 lactone concentrations disappeared at the same rate regardless of whether cells were present, initially falling to reach equilibrium with the hydroxy acid by 4 h. Indeed, changes in intracellular lactone concentrations followed closely chemical stability profiles in media. Similar patterns of cellular retention and chemical degradation were observed with CPT. CONCLUSION: The major determinant of drug accumulation in three diverse cell line phenotypes was lactone chemical stability in culture medium. PMID- 11935212 TI - Phase I and pharmacokinetic study of SPI-77, a liposomal encapsulated dosage form of cisplatin. AB - PURPOSE: To investigate the safety and pharmacokinetics of a new liposomal formulation of cisplatin, SPI-77, in patients with advanced malignancies. PATIENTS AND METHODS: Patients with histologically proven malignancies not amenable to other treatment were eligible for this study. The starting dose of SPI-77 (cisplatin in Stealth liposomes) was 40 mg/m(2) administered every 4 weeks in a 2-h infusion, and doses were escalated up to 420 mg/m(2). Pharmacokinetic monitoring was performed in all patients and samples were analysed for platinum content by atomic absorption spectroscopy. Platinum-DNA (Pt-DNA) adduct levels in leucocytes (white blood cells, WBC) and tumour tissue were quantified using a sensitive (32)P-postlabelling assay. RESULTS: A total of 27 patients were accrued. The main toxicities observed were infusion-related reactions, which could be prevented by lowering the initial infusion rate, and anaemia. The pharmacokinetics of SPI-77-derived platinum were strikingly different from standard cisplatin. Free platinum levels in plasma ultrafiltrate samples were undetectable at the lowest dose levels (40 and 80 mg/m(2)), and low but highly variable at higher doses of SPI-77. Plasma pharmacokinetics of total platinum were linear with small interpatient variability. The total body clearance of SPI 77 varied from 14 to 30 ml/h and was significantly lower than reported clearance values for cisplatin of 20 l/m(2) per h, due to the slow release of cisplatin from the liposomes. Pt-DNA adduct levels in WBC ranged from 0.02 to 4.13 fmol/microg DNA for intrastrand Pt-GG (guanine-guanine) adducts and from 0.02 to 1.27 fmol/microg DNA for intrastrand Pt-AG (adenosine-guanine) adducts, which is more than tenfold lower than after administration of a comparable dose of non liposomal cisplatin. In tumour samples obtained from two patients treated at the highest dose-levels, relatively low levels of Pt-DNA adducts were observed. CONCLUSIONS: The results of this phase I trial show that the pharmacokinetic behaviour of cisplatin is significantly altered by its encapsulation in Stealth liposomes. The pharmacokinetics of SPI-77 are mainly dominated by the liposomal properties, resulting in high cholesterol concentrations and relatively low concentrations of (free) platinum in plasma, WBC and tumour tissue, which may explain the observed differences between the toxicity profiles of SPI-77 and cisplatin. PMID- 11935213 TI - Antitumor activity of combinations of anti-HER-2 antibody trastuzumab and oral fluoropyrimidines capecitabine/5'-dFUrd in human breast cancer models. AB - The anti-HER-2 antibody, trastuzumab (Herceptin), and the oral fluoropyrimidine, capecitabine (Xeloda), are both effective in breast cancer with different modes of action and toxicity profiles. Therefore, the efficacy in combination therapy of these agents for the treatment of HER-2-overexpressing breast cancer was of interest. An antagonistic interaction in vitro between trastuzumab and 5 fluorouracil (5-FUra) in combination has previously been reported. In the same study, the in vivo antitumor activity of this combination was investigated. However, the results were inconclusive since 5-FUra at the dose used had sufficient antitumor efficacy as a single agent and therefore it was not possible to clarify the potential difference between 5-FUra alone and the combination. In the present study, we investigated the efficacy of trastuzumab in combination with capecitabine or its intermediate metabolite 5'-dFUrd in HER-2-overexpressing human breast cancer cell lines and xenograft models. In vivo antitumor activity of the combination was at least additive, in terms of tumor growth inhibition and tumor growth delay, in human breast cancer models KPL-4 and BT-474. The combination treatment in vivo was superior to the treatment with either single agent alone, even though in vitro treatment with trastuzumab and 5'-dFUrd/5-FUra showed antagonistic antiproliferative activity in these cell lines. The reason for the discrepancy between the in vivo and in vitro results was not clarified. However, observed additive in vivo antitumor activity clearly indicates that the clinical efficacy of the combination of trastuzumab and capecitabine/5'-dFUrd against HER-2-overexpressing breast cancer is worth investigating. PMID- 11935214 TI - Apoptotic and genotoxic effects of a methyl sulfonate ester that selectively generates N3-methyladenine and poly(ADP-ribose) polymerase inhibitors in normal peripheral blood lymphocytes. AB - Selective N3-adenine methylation represents a novel strategy for tumors with a phenotype of poor responsiveness to a number of anticancer agents currently used in the clinic. Resistance to N3-methyladenine-inducing agents, such as MeOSO(2)(CH(2))(2)-lexitropsin (Me-Lex), is due to high levels of N-methylpurine glycosylase (MPG). However, tumor cells with high MPG activity can be rendered susceptible to Me-Lex using poly(ADP-ribose) polymerase-1 (PARP-1) inhibitors. PURPOSE: To evaluate the potential toxicity of Me-Lex, used as single agent or combined with PARP-1 inhibitors, in normal peripheral blood lymphocytes (PBL). METHODS: PBL either resting or activated with phytohemagglutinin (PHA), obtained from healthy donors, were treated with graded concentrations of Me-Lex with or without PARP-1 inhibitor (3-aminobenzamide, AB, or NU1025, NU). MPG activity, apoptosis and sister chromatid exchanges (SCE) were evaluated. RESULTS: (a) Me Lex was cytotoxic mainly in PHA-activated PBL with low MPG activity; (b) combined treatment with Me-Lex and AB induced apoptotic effects as early as 24 h after drug exposure both in non-stimulated and PHA-activated PBL. When concentrations of PARP-1 inhibitors (25 microM NU and 4 m M AB) that produced a twofold increase in Me-Lex cytotoxicity in tumor cells were compared, NU induced a less-pronounced increase in apoptosis in PBL treated with Me-Lex; (c) Me-Lex at concentrations that allowed cytogenetic analysis did not induce a significant number of SCE; (d) PARP-1 inhibitors provoked a dose-dependent increase in SCE, but 25 microM NU was devoid of genotoxic effects and did not significantly increase SCE in PBL treated with Me-Lex. CONCLUSIONS: Me-Lex showed preferential cytotoxicity against mitogen activated PBL. Our results also indicated that for each PARP-1 inhibitor it is necessary to define the concentration devoid of genotoxic effects in normal cells, but still capable of enhancing the efficacy of DNA-damaging agents in tumor cells. PMID- 11935215 TI - Effect of renal impairment on the pharmacokinetics and tolerability of capecitabine (Xeloda) in cancer patients. AB - PURPOSE: The primary objective of this study was to investigate the influence of renal impairment on the pharmacokinetics of capecitabine and its metabolites in cancer patients. Capecitabine (Xeloda) is an orally administered precursor of 5' deoxy-5-fluorouridine (5'-DFUR), which is preferentially activated to 5 fluorouracil (5-FU) in tumors. METHODS: A total of 27 patients were enrolled, of whom 24 were evaluable for pharmacokinetics (6 with normal renal function, 8 with mild, 6 with moderate, and 4 with severe renal impairment at baseline). Patients received capecitabine orally at the standard dosing regimen (1250 mg/m(2) capecitabine twice daily for 2 weeks followed by a 1-week rest period). On study days 1 and 14, blood samples were collected to evaluate the pharmacokinetics of capecitabine and its metabolites. The relationship between the area under the plasma concentration-time curve (AUC) and creatinine clearance (CL(CR)) was assessed by log-linear regression analysis. RESULTS: The primary pharmacokinetic parameter with respect to the effect of renal dysfunction was systemic exposure to 5'-DFUR, 5-FU and FBAL determined on study day 14. Renal impairment led to an increase in the systemic exposure to 5'-DFUR and FBAL (23% and 109% increase in AUC, respectively) for a 50% reduction in CL(CR). By contrast, renal impairment may lead to decreased exposure to 5'-DFCR. There was no evidence for an effect of renal impairment on systemic exposure to 5-FU or capecitabine. Renal impairment did not have a major effect on peak concentration (C(max)) or elimination half life (t(1/2)) of capecitabine, 5'-DFCR, 5'-DFUR, and 5-FU. However, in the case of FBAL, moderate or severe renal impairment caused up to a twofold increase in C(max) and prolongation of t(1/2). All patients with severe renal impairment (four patients) had drug-related grade 3 or 4 adverse-events (AEs) and serious AEs. Patients with moderate renal impairment experienced a similar number of grade 3 or 4 AEs (six of nine patients) but had a higher incidence of serious AEs (three of nine patients) when compared with those with normal renal function (four of six patients and one of six patients, respectively). A similar effect was seen in patients with mild renal dysfunction (grade 3 or 4 AEs in four of eight patients; serious AEs in three of eight patients). The relationship between systemic exposure to capecitabine or its metabolites and safety was investigated using logistic regression. This exploratory analysis showed a strong positive relationship between AUC of 5'-DFUR and treatment-related grade 3 or 4 AEs, whereas there was no relationship with exposure to capecitabine, 5'-DFCR, 5-FU or FBAL. CONCLUSIONS: Renal impairment has no effect on the pharmacokinetics of capecitabine or 5-FU, but leads to an increase in the systemic exposure to 5' DFUR and FBAL. However, only the AUC of 5'-DFUR is correlated with safety. Based on the safety results in patients with severe renal impairment, a dose modification cannot be recommended for these patients and they should not be treated with capecitabine. Additional data from the clinical safety database and pharmacokinetic results from the present study support the recommendation that patients with moderate renal impairment should be treated with 75% of the recommended standard starting dose to achieve systemic exposure comparable to that in patients with normal renal function. PMID- 11935216 TI - Influence of oxaliplatin on 5-fluorouracil plasma clearance and clinical consequences. AB - The influence of oxaliplatin (OXA) on 5-fluorouracil (5-FU) plasma clearance was investigated. PATIENTS AND METHODS: A group of 29 patients with advanced colorectal cancer refractory to prior weekly 8-h 5-FU infusion plus bolus folinic acid (FA), received the same combination plus OXA at 130 mg/m(2) every 3 weeks, OXA plus 5-FU plus FA on day 1, and 5-FU plus FA on days 8 and 15. Steady-state 5 FU concentrations in plasma were measured weekly and 5-FU clearance was calculated. Both before and after the addition of OXA, the 5-FU dose was individually adjusted according to the pharmacokinetic follow-up (target steady state plasma concentrations 2.5-3 mg/l). RESULTS AND DISCUSSION: A total of 122 OXA-containing infusions and 338 5-FU plus FA infusions were given and the median number of infusions per patient was 4 (2-9) and 10 (5-28), respectively. 5-FU plasma clearance was significantly decreased on days 8 and 15 when compared with the value on day 1 and with the values before OXA introduction using a direct paired comparison (2.36 and 2.31 l/min, respectively, vs 3.12 and 3.05 l/min; P<10(-5)). Of 25 evaluable patients, 6 had an objective response after the introduction of OXA (24% objective response rate, 95% confidence interval 9.4 45%). CONCLUSION: OXA reduces 5-FU plasma clearance for 15 days. This may be a factor in the synergy between the two drugs. It is not linked to dihydropyrimidine dehydrogenase inhibition. Implications for drug schedules in clinical practice are discussed. PMID- 11935217 TI - Relationships between the in vitro cytotoxicity and transport characteristics of pirarubicin and doxorubicin in M5076 ovarian sarcoma cells, and comparison with those in Ehrlich ascites carcinoma cells. AB - PURPOSE: We sought to determine whether the de novo resistance of M5076 ovarian sarcoma cells, which show sensitivity to pirarubicin (THP), to doxorubicin (DOX) is due to differences in the transport characteristics between THP and DOX, and the results were compared with those for drug-sensitive Ehrlich ascites carcinoma cells. METHODS: The in vitro cytotoxicity of the drugs was assessed by means of the tetrazolium dye assay. Transport experiments were performed by the rapid centrifugation method. RESULTS: In an in vitro cytotoxicity experiment, M5076 cells showed lower sensitivity to DOX than to THP, and the cytotoxicity of THP and DOX toward M5076 cells was lower than toward Ehrlich cells, and these results were similar to those of an in vivo experiment. This was due to the much lower expression of topoisomerase II in M5076 cells than in Ehrlich cells. The amount of intracellular DOX was found to be significantly lower than that of THP in both cell types, and furthermore, little free intracellular DOX was observed in M5076 cells, indicating that the low sensitivity of M5076 cells to DOX was partially a result of the low amount of intracellular DOX. There was no difference in the efflux rate, but there was an apparent difference in the uptake efficiency of the carrier between THP and DOX. CONCLUSIONS: These findings suggest that the cytotoxicities of THP and DOX toward M5076 and Ehrlich cells depend, at least in part, on the uptake efficiency of the carrier. PMID- 11935218 TI - Preclinical study of dolastatin-10 in dogs with spontaneous neoplasia. AB - PURPOSE: To evaluate the short-term adverse effects of administration of dolastatin-10 (Dol-10) to dogs with spontaneously occurring malignant tumors. METHODS: A total of 34 tumor-bearing dogs were given Dol-10 as a rapid intravenous bolus every 14 days at starting dosages ranging from 200 to 350 microg/m(2). Acute and short-term adverse effects, antitumor response, and duration of response were characterized. RESULTS: The maximum tolerated dose varied greatly from patient to patient, but a reasonable starting dose for further studies was established at 300 microg/m(2). The median number of treatments per dog was 2 (range 1 to 17). Granulocytopenia was the dose-limiting toxicity. The overall response rate was 3%, consisting of a complete and durable (30 months) response in a dog with high-grade malignant lymphoma that was refractory to standard therapy. Two minor or transient responses were observed, and two dogs experienced disease stabilization for 8 and 16 weeks. CONCLUSIONS: Dol-10 appears to be well tolerated in tumor-bearing dogs at doses approaching those tolerated by humans. The clinical activity observed in dogs with non Hodgkin's lymphoma warrants further investigation. PMID- 11935219 TI - Neoadjuvant treatment of locally advanced carcinoma of the uterine cervix with epirubicin, paclitaxel and cisplatin. AB - PURPOSE: The present study was conducted to explore whether neoadjuvant chemotherapy with a combination of epirubicin, paclitaxel and cisplatin could improve the operability and pathological response rate in locally advanced cervical cancer patients. METHODS: Between April 1996 and July 2000, 42 patients with carcinoma of the uterine cervix, FIGO stage Ib(2)-IVa, were treated with two or three 21-day cycles of an epirubicin 100 mg/m(2), paclitaxel 175 mg/m(2), cisplatin 100 mg/m(2) regimen. RESULTS: All patients were eligible for evaluation of toxicity and response. A total of 92 courses of therapy were administered. Three patients had a 20% reduction from the starting dose due to haematological toxicity. Grade 3-4 leukopenia was observed in 15% of cycles, requiring G-CSF support in half of them. Major non-haematological toxicity consisted of grade 3 alopecia (100%), and grade 3 nausea and vomiting (40%). A total of 33 clinical responses (78.5%, 95% CI 63.8-93.2) were recorded, 8 complete responses (CR) and 25 partial responses (PR). Of the 42 patients, 32 (76.2%) underwent radical surgery. At pathological examination 8 complete or microscopic pathological responses, 17 PRs, and 9 patients with stable disease were observed. The median follow-up time was 17 months for the 42 patients enrolled (range 3-62 months). Among the patients submitted to radical surgery, five recurrences were observed, with a median disease-free survival of 47 months. Median overall survival had not been reached at the time of this report. These results appear to be in the range reported for other neoadjuvant cisplatin-based regimens not including paclitaxel. CONCLUSIONS: Neoadjuvant chemotherapy with the epirubicin, paclitaxel and cisplatin combination followed by radical surgery proved to be a safe and effective approach to advanced cervical cancer. PMID- 11935220 TI - Composition of the spindle pole body of Saccharomyces cerevisiae and the proteins involved in its duplication. AB - The microtubule-organizing center (MTOC) is the organelle that manages the distribution of microtubule filaments in the cell. It includes such centers of microtubule organization as basal bodies, nucleus-associated bodies, spindle pole bodies and centrosomes. The centrosome is the MTOC in most animal cells. Despite the numerous studies that have been published on this organelle over the past 100 years, the protein composition of the animal centrosome remains poorly defined. In order to study the MTOC, many researchers have chosen the budding yeast Saccharomyces cerevisiae as a model organism. This review summarizes the budding yeast MTOC, referred to as the spindle pole body (SPB), including its structure and function, its components and the factors involved in its duplication. Electron microscopy and high-voltage electron tomographic analysis of this organelle have contributed to the understanding of its morphology, whereas analysis by matrix-assisted laser desorption/ionization has played a major role in identifying many of the spindle pole components. In addition, genetic and biochemical studies have revealed the functional and physical relationships of certain spindle pole components and proteins important for SPB duplication. PMID- 11935221 TI - Sequence-based approach for identification of cell wall proteins in Saccharomyces cerevisiae. AB - Open reading frames (ORFs) in the genome of Saccharomyces cerevisiae were screened for cell wall proteins and extracellular proteins, using an in silico sequence analysis combined with biochemical examination. The selection criteria used in the sequence analysis were the presence of a signal sequence for secretion and the absence of any targeting and retention signal to/in intracellular components. By using the PSORT II program, 163 ORFs/proteins were selected as potential extracellular proteins, including cell wall proteins. Of these, 51 ORFs/proteins of unknown localization and more than 120 amino acids in size were further studied on their cellular localization. A hemagglutinin (HA) epitope was inserted in the most C-terminus of each protein and the resulting HA tagged protein was expressed under the authentic promoter in yeast cells. Out of the 51 constructs, 35 gave protein bands on Western blots. Examination of proteins in fractionated samples identified 11 extracellular proteins; six proteins that were weakly associated with the cell wall and five proteins that were relatively tightly associated with the cell wall. PMID- 11935222 TI - Functional interactions between the VRP1-LAS17 and RHO3-RHO4 genes involved in actin cytoskeleton organization in Saccharomyces cerevisiae. AB - The RGD1 gene from Saccharomyces cerevisiae, which encodes a GTPase-activating protein for the Rho3 and Rho4 small G proteins, exhibits synthetic lethality with the VRP1 and LAS17 genes. Their products are proline-rich proteins that interact with both actin and myosins to ensure polarized growth. By testing functional links, we found that the VRP1 and LAS17 genes are potent suppressors of the rho3Delta mutation. In particular, they restore the polarization of actin patches in rho3Delta cells. Moreover, the vrp1Delta and las17Delta mutations were found to display a similar pattern of genetic interactions with specific actin-linked genes. These mutations also increase the sensitivity to activated forms of both Rho3p and Rho4p. These data support our working model, in which the VRP1 and LAS17 genes define a cellular complex that works in concert with the RHO3-RHO4 signaling pathway in yeast polarized growth. In addition, other observations lead us to propose that Rvs167p may act as a linking protein between the two cellular elements. PMID- 11935223 TI - Nep1p (Emg1p), a novel protein conserved in eukaryotes and archaea, is involved in ribosome biogenesis. AB - The essential Saccharomyces cerevisiae gene YLR186w [ NEP1 (nucleolar essential protein 1), also recently named EMG1 (essential for mitotic growth 1)] is highly conserved in eukaryotes and archaea. In S. cerevisiae, Nep1p is localized in the nucleus with a rod-shaped morphology, which first suggested a spindle/microtubule association for ScNEP1 (S. cerevisiae NEP1). However, in Candida albicans and in HeLa cells, Nep1p is a protein of the nucleoli. CaNEP1 (C. albicans NEP1) and HsNEP1 (Homo sapiens NEP1) heterologously complement the essential phenotype in a S. cerevisiae nep1 deletion mutant. However, the ScNEP1 spindle/microtubule phenotype is not found with HsNEP1 and CaNEP1, which shows that the spindle/microtubule association is specific for ScNep1p and that it is not the primary essential function of Nep1p. A temperature-sensitive ScNEP1 (ts1) allele was isolated and revealed a strongly increased sensitivity to paromomycin, a translational inhibitor which binds to RNA, indicating that ribosome biogenesis within the nucleolus is probably affected. This was confirmed by polysome profile and ribosomal subunit analysis, showing a ribosomal subunit imbalance with a decrease in 40S subunits, due to reduced amounts of native 18S rRNA. Furthermore, ScSAM2, encoding S-adenosylmethionine synthetase, was isolated as a multicopy suppressor of the ScNEP1 (ts1) mutant allele and medium supplementation with S adenosylmethionine restored its growth. For the first time, these results identified a novel protein with an essential function in ribosomal biogenesis which directly or indirectly interferes with a methylation reaction during the early steps of pre-rRNA processing necessary for the generation of 40S ribosomal subunits. PMID- 11935224 TI - A Penicillium chrysogenum gene ( aox) identified by specific induction upon shifting pH encodes for a protein which shows high homology to fungal alcohol oxidases. AB - Parallel cultures of Penicillium chrysogenum were grown in controlled bioreactors under conditions of penicillin production and one was shifted from the initial pH 6.0 to pH 8.0. RNA was isolated from both cultures and used for a differential hybridization experiment to identify genes specifically induced upon this pH shift. About 2,000 plaques of a cDNA library constructed from pH 8.0 material were screened with a pH 8.0 to pH 6.0 subtracted probe. Two independent clones of which the RNA was highly abundant in pH 8-shifted material and essentially not present in pH 6 material were isolated. Both clones were found to belong to one specific gene, which could be identified by sequence homology as an alcohol oxidase. The identified aox gene encoded for a peptide of 666 amino acids, interrupted by nine introns; and it showed high homology (>65% identity) to alcohol oxidases from Cladosporium fulvum and the methanol-utilizing yeasts Candida boidinii, Hansenula polymorpha (now Pichia angusta) and P. pastoris. The transcription start was identified by primer extension analysis. Southern analysis revealed that related genes are widely distributed among fungal species. PMID- 11935225 TI - The nuclear genome of a Drosophila mutant strain increases the frequency of rearranged mitochondrial DNA molecules. AB - We studied a mutant strain of Drosophila subobscura, in which 80% of the mitochondrial genomes (mtDNA) have lost over 30% of the coding region. The mutation is stable and is transmitted identically to offspring. The putative role of the mutant nuclear genome in the production of rearranged mtDNA was investigated using reciprocal crosses, to place the mitochondria of the wild strain in a mutant nuclear context. Nested PCR was used to screen for rearrangements in different regions of mtDNA; and rearrangements were detected in some individuals from the F6 generation. The frequency of these deleted mtDNAs then increased progressively in the population; and they were present in nearly all individuals in the F11 generation. They were not transmissible. Direct repeats were present at the deletion boundaries. These mutated genomes disappeared on reversion to a wild-type nuclear genome. Deletions were detected in a very small fraction of the wild population (0.7% of individuals). The mutant nuclear genome therefore does not promote a particular deletion but increases the frequency of different mtDNA rearrangements. The potential involvement of different candidate nuclear genes is discussed. PMID- 11935226 TI - Visual field structure in the Empress Leilia, Asterocampa leilia (Lepidoptera, Nymphalidae): dimensions and regional variation in acuity. AB - Male Empress Leilia butterflies ( Asterocampa leilia) use a sit-and-wait tactic to locate mates. To see how vision might influence male behavior, we studied the morphology, optics, and receptor physiology of their eyes and found the following. (1) Each eye's visual field is approximately hemispherical with at most a 10 degrees overlap in the fields of the eyes. There are no large sexual differences in visual field dimensions. (2) In both sexes, rhabdoms in the frontal and dorsal ommatidia are longer than those in other eye regions. (3) Interommatidial angles are smallest frontally and around the equator of the eye. Minimum interommatidial angles are 0.9-1 degrees in males and 1.3-1.4 degrees in females. (4) Acceptance angles of ommatidia closely match interommatidial angles in the frontal region of the eye. We conclude that vision in these butterflies is mostly monocular and that males have more acute vision than females, especially in the frontal region (large facets, small interommatidial angles, small acceptance angles, long rhabdoms, and a close match between interommatidial angles and acceptance angles). This study also suggests that perched males direct their most acute vision where females are likely to appear but show no eye modifications that appear clearly related to a mate-locating tactic. PMID- 11935227 TI - Experimental modification of stepping course in spontaneously initiated locomotor behavior in the crayfish Procambarus clarkii Girard. AB - The stepping course in spontaneously initiated walking of crayfish was quantitatively analyzed using a spherical treadmill system. In complete darkness, some animals stepped either forward or backward at random whereas others showed individually a consistent tendency of stepping in a specific direction although no external sensory cue was provided. The tendency was statistically significant and invariable for at least 6-8 h. When a light stimulus was present in front of the animal, the stepping course tended to be backward or curved forward to avoid the stimulus. Either in complete darkness or in the presence of a light stimulus, the animal's tendency to step in a specific direction could be modified experimentally by applying electrical stimulation to a part of the animal body upon stepping in the preferred direction. The newly acquired tendency of stepping direction could be retained for 6 h and modified again by a similar procedure of electric stimulation. Both before and after modification of the stepping course tendency, animals seldom changed their stepping direction once the walking was initiated. These findings suggest that the stepping course in spontaneously initiated walking is significantly affected by animal's previous experience and could be predetermined at the onset of walking. PMID- 11935228 TI - Auditory stimulus-induced changes in immediate-early gene expression related to an inborn perceptual predisposition. AB - Auditory-induced expression of the immediate-early gene ZENK was examined in 18 brain areas of domestic chicken and Japanese quail subjects with no previous exposure to parental vocalizations. After one 30-min exposure (approximately 120 calls) within 24 h of hatching to either the chicken or quail maternal call, paired subjects from each species showed significantly more intense levels of ZENK staining to conspecific (own species) calls. Single brain areas did not show consistent, large differences in ZENK expression across all subject pairs. Instead, a majority of brain areas exhibited a small conspecific stimulus-induced staining advantage in each species. The species differed in the quantitative distribution of ZENK responses among brain areas; qualitative patterns of call induced staining exhibited both similarities and differences. The results suggest shared neural correlates and potential developmental/evolutionary differences in congenital brain responses to biologically significant auditory stimuli in naive individuals of the two species. PMID- 11935229 TI - Control of obstacle climbing in the cockroach, Blaberus discoidalis. I. Kinematics. AB - An advantage of legged locomotion is the ability to climb over obstacles. We studied deathhead cockroaches as they climbed over plastic blocks in order to characterize the leg movements associated with climbing. Movements were recorded as animals surmounted 5.5-mm or 11-mm obstacles. The smaller obstacles were scaled with little change in running movements. The higher obstacles required altered gaits, leg positions and body posture. The most frequent sequence used was to first tilt the front of the body upward in a rearing stage, and then elevate the center of mass to the level of the top of the block. A horizontal running posture was re-assumed in a leveling-off stage. The action of the middle legs was redirected by rotations of the leg at the thoracal-coxal and the trochanteral-femoral joints. The subsequent extension movements of the coxal trochanteral and femoral-tibial joints were within the range seen during horizontal running. The structure of proximal leg joints allows for flexibility in leg use by generating subtle, but effective changes in the direction of leg movement. This architecture, along with the resulting re-direction of movements, provides a range of strategies for both animals and walking machines. PMID- 11935230 TI - Control of climbing behavior in the cockroach, Blaberus discoidalis. II. Motor activities associated with joint movement. AB - Deathhead cockroaches employ characteristic postural strategies for surmounting barriers. These include rotation of middle legs to re-direct leg extension and drive the animal upward. However, during climbing the excursions of the joints that play major roles in leg extension are not significantly altered from those seen during running movements. To determine if the motor activity associated with these actions is also unchanged, we examined the electromyogram activity produced by the slow trochanteral extensor and slow tibial extensor motor neurons as deathhead cockroaches climbed over obstacles of two different heights. As they climbed, activity in the slow trochanteral extensor produced a lower extension velocity of the coxal-trochanteral joint than the same frequency of slow trochanteral extensor activity produces during horizontal running. Moreover, the pattern of activity within specific leg cycles was altered. During running, the slow trochanteral extensor generates a high-frequency burst prior to foot set down. This activity declines through the remainder of the stance phase. During climbing, motor neuron frequency no longer decreased after foot set-down, suggesting that reflex adjustments were made. This conclusion was further supported by the observation that front leg amputees generated even stronger slow trochanteral extensor activity in the middle leg during climbing movements. PMID- 11935231 TI - Impairment of central pattern generation in Drosophila cysteine string protein mutants. AB - The cysteine string proteins are integral synaptic vesicle proteins, critical for fast, Ca2+ -regulated exocytosis. Drosophila larvae with null mutations for the cysteine string protein (csp) gene have temperature-sensitive impairments of neurotransmission and presynaptic calcium removal at the neuromuscular junction. Using the larval Drosophila preparation to examine central pattern generation, we characterized the temperature sensitivity of locomotor patterns in wildtype and csp mutant larvae. Intraburst frequency of motoneuronal activity reached 100 Hz and was sufficiently high to rescue the temperature-sensitive synaptic failure in the mutant. Nevertheless, we show that deletion of the csp gene resulted in a severe deficiency in the generation of coordinated larval motor rhythms. Csp mutants that could generate patterned motor activity had slower, poorly coordinated rhythms with altered temperature sensitivity. We conclude that the temperature-sensitive paralysis characteristic of csp mutants is not a direct result of synaptic failure at neuromuscular junctions, as might be expected, but is the result of a failure of locomotor circuit operation at a higher integrative level. PMID- 11935233 TI - Surgically treated cerebellitis. PMID- 11935235 TI - Epidemiology of pediatric central nervous system tumor: a meta-analysis or review? PMID- 11935237 TI - Progressive ventricular enlargement in the absence of high ventricular pressure in an experimental neonatal rat model. AB - OBJECT: In the present study, we compared ventricular pressures (VP) and the progression of ventricular enlargement in a new experimental neonatal hydrocephalus model, to gain an understanding of how communicating hydrocephalus progresses. METHODS: Kaolin was injected into the subarachnoid space at the cranial convexity of neonatal rats. Gross examination was performed on the 3rd, 5th and 7th days, and ultrasonographic examination on the 15th day, and at the end of the 1st and 2nd months following the kaolin application. Ventricular size indexes (VSI) were calculated in the case of a large ventricular dilatation. VPs were assessed on the 15th day, and at the end of the 1st and 2nd months, with a computerized data acquisition system. CONCLUSIONS: In the 1st and 2nd months VSIs were significantly higher than in control rats on the 15th day after kaolin administration. VP on the 15th day was significantly increased compared with that in control rats. VP in the 1st month was still high, but had subsided. In the 2nd month VP was not increased over control. In the late stages, the progression of infantile communicating hydrocephalus is not related to VP levels. PMID- 11935238 TI - Distal slit valve and clinically relevant CSF overdrainage in children with hydrocephalus. AB - INTRODUCTION: Distal slit valve (DSV) is a system designed for the treatment of hydrocephalus. It has been assumed that, by dispensing with an anti-siphon (AS) mechanism, the DSV induces a set of clinical symptoms associated with fluid overdrainage in patients. Nonetheless, there is no published evidence to support this assumption. MATERIALS AND METHODS: Thus, to determine whether such an association is valid, we reviewed the records of 101 hydrocephalic patients (150 procedures) who had DSVs placed at our institution. The records of 40 hydrocephalic patients (69 procedures) in whom anti-siphon devices (AS) were placed were also reviewed. RESULTS: One DSV patient presented with slit ventricle syndrome (SVS) and low intracranial pressure (ICP). No DSV patients had postoperative subdural collection. One AS patient had a postoperative subdural collection. Thirty-one DSV patients (31%) each required one revision, and 8 (8%) required more than one revision. Twelve AS patients (30%) required one revision and 8 AS patients (20%) required more than one revision. No significant differences were found between the DSV and AS groups in number of revisions, infections or overdrainage. CONCLUSION: We did not find a direct correlation between clinically relevant CSF overdrainage and DSV. PMID- 11935239 TI - Calcific deposits on degraded shunt catheters: long-term follow-up of V-P shunts and late complications in three cases. AB - OBJECT: Three cases with late complications and nine cases without late complications are presented to demonstrate the long-term results of ventriculoperitoneal (V-P) shunting, particularly those concerning degradation and mineral deposits of shunt catheters. METHODS: Plain X-rays were taken in every case to detect any calcification. The catheters removed following late complications were examined in a scanning electron microscope (SEM). Spectroscopy and conventional histology were also performed. CONCLUSION: Routine histology, spectroscopy, and SEM revealed that the mineral deposits consisted of hydroxyapatite. Plain X-ray and operative findings showed that the most extensive calcification was present in the neck, where the catheters were subject to heavy mechanical stress. No calcification was detected on catheters that did not contain barium particles. Our findings indicate that mechanical stress contributes to the process of degradation, and that barium sulfate admixed with silicone during the manufacturing process might accelerate late complications owing to the formation of cracks in the catheters and by enhancing the nucleation rate. PMID- 11935240 TI - Ventricular catheter placement in children with hydrocephalus and small ventricles: the use of a frameless neuronavigation system. AB - OBJECT: Accurate placement of ventricular catheters in children with small ventricles can be difficult. Too often, shunt catheters are misplaced with regard to optimal position and trajectory. The objective of this study was to determine whether neuronavigation-guided free-hand placement of ventricular catheters is an effective adjunct for children with hydrocephalus and small ventricles. METHODS: Nine children with hydrocephalus (ages 1-12 years) participated in this study. Four children were diagnosed as suffering from slit ventricle syndrome and 5 children had small to mildly dilated ventricles. Of the 9 shunted children, 6 underwent previous shunt placements, and 1 child previously underwent an endoscopic third ventriculostomy. In 8 children the primary procedure was insertion of ventricular catheters using a frameless neuronavigation system. In 1 child, the neuronavigation system was used after failure to insert the ventricular catheter using a standard technique. All children showed significant improvement of their symptoms and signs following the procedure and none of them required shunt revision during the follow up period (mean 8+/-5 months). CONCLUSION: The usage of a neuronavigation system is safe and may be beneficial for optimal positioning and trajectory of ventricular catheters in children with small ventricles or an abnormal ventricular anatomy. PMID- 11935241 TI - Pediatric brain tumors: statistics of SNUH, Korea (1959-2000). AB - OBJECTIVE: To investigate the age, gender, location and pathologic diagnosis of brain tumors in pediatric patients (less than 16 years old), we reviewed 677 patients who were operated on at the Department of Neurosurgery, Seoul National University Hospital (SNUH) during the period from January 1959 to June 2000. PATIENTS AND METHODS: Only tumors for which pathologic specimens were available were included. Tumors of bone origin, purely extradural mass, nontumorous cystic lesions, and vascular malformations were excluded. The mean age of the 677 patients was 7.8 years and the gender ratio (male-to-female ratio), 1.4:1. Supratentorial tumors (60.0%) were more common than infratentorial tumors (39.1%). Pathological examination showed that the most common tumors were astrocytic tumors (25.7%), medulloblastomas (17.9%), craniopharyngiomas (12.0%), germ cell tumors (11.2%), supratentorial primitive neuroectodermal tumors (ST PNETs) (5.5%), and neuronal tumors (5.2%). Choroid plexus tumors and ependymal tumors occurred more frequently in early childhood, while pituitary adenomas and non-teratomatous germ cell tumors occurred more frequently in older children. While most tumors were more predominant in males, oligodendroglial tumors and pituitary adenomas were more predominant in females. CONCLUSIONS: The relative incidences of germ cell tumors, neuronal tumors, and oligodendroglial tumors increased after the introduction of magnetic resonance imaging (MRI). In contrast, the incidence rates of medulloblastomas and ependymal tumors decreased. PMID- 11935242 TI - Survival of children under 3 years old with medulloblastoma: a study from the Mexican Cooperative Group for Childhood Malignancies (AMOHP). AB - BACKGROUND: The prognosis of medulloblastoma in children under 3 years of age is poor. METHODS: A retrospective analysis was performed to evaluate children under 3 years of age with medulloblastoma. Overall survival (OS) and progression-free survival (PFS) were assessed in children with and without metastasis. RESULTS: A total of 534 children were diagnosed with medulloblastoma during the study period, 49 (9.1%) of whom were under 3 years of age and were evaluated. Their ages ranged from 5 to 35 months with a mean of 18.5 months. In 39 (79.6%) of these patients the tumors were staged as T3M0 or under, while 10 (20.4%) had metastasis at diagnosis. The OS was 38% and PFS 37% in the whole series, while PFS was 32% in those with metastasis and 40% in those without ( P=0.78). For those who received radiotherapy the PFS was 62%, and in those not treated with radiotherapy PFS was nil ( P=0.0001). When the children were divided into those who received surgical treatment plus chemotherapy and those who received surgery plus radiotherapy plus chemotherapy, the PFS was nil and 66%, respectively ( P=0.00001). CONCLUSION: Because of the high morbidity of radiotherapy in children under 3 years old, surgery continues to be the basis of improved prognosis, followed by chemotherapy. PMID- 11935243 TI - Hemispheric cerebral gliomas in children with NF1: arguments for a long-term follow-up. AB - BACKGROUND: Neurofibromatosis type 1, or peripheral neurofibromatosis, is the most form of common phakomatosis. In some instances, it can be associated with tumors of the central nervous system. CASE REPORT: We describe four cases of hemispheric cerebral glioma in children with NF1. Two of these tumors were pilocytic astrocytomas and two, ependymomas. We reviewed the literature on hemispheric cerebral gliomas in NF1 patients in an attempt to find characteristic features of and some explanations for these lesions. CONCLUSION: We conclude that surgical resection of these tumors should be proposed whenever possible, with due consideration for their progressive nature and the uncertainties about malignancy. We advocate yearly clinical and neuroradiological follow-up over a long period in NF1 children, since they may develop additional CNS tumors during their lifetime. PMID- 11935244 TI - Functional investigation in children with spina bifida -- measured by the Pediatric Evaluation of Disability Inventory (PEDI). AB - OBJECT: The purpose of this study was to investigate the functional performance in children with spina bifida, using the Pediatric Evaluation of Disability Inventory (PEDI). A comparison was also made between the functional performance of children with myelomeningocele (MMC) and lipomyelomeningocele (LMC). METHODS AND RESULTS: Twenty-six children with MMC and 37 cases with LMC were enrolled in this study. There was no significant difference in gender, age or method of birth delivery between these two study groups. The normative standard scores of PEDI for children with spina bifida were all below the mean standard scores found in non-disabled children, but still within the average range, with exception of mobility score in the MMC group. Patients' neurological level, walking ability, intelligence, independence in activities of daily living, and bowel function showed significant correlation with the PEDI scores ( P<0.05) in the children with MMC. In comparing these two disease groups, the MMC had higher neurological level, poorer walking ability and poorer bladder and bowel function than did the LMC ( P<.05). The former also had significantly lower PEDI scores in the mobility and social function domains. PEDI provided valuable information regarding overall functional performance in daily life for children with spina bifida. CONCLUSION: This information is extremely important for planning a comprehensive and individualized rehabilitation program. PMID- 11935245 TI - The impact of initial management on the outcome of children with severe head injury. AB - BACKGROUND: Neurotrauma is one of the most important causes of death in the pediatric age group. Head injury is responsible for both primary and secondary brain damage. The outcome of children with brain injury depends on the nature of the primary damage and on how well secondary brain damage can be limited. Appropriate critical care management at the scene and in hospital can prevent secondary brain damage that would otherwise result from cerebral reactions. Hypoxia and hypotension are major early complications of aggravation of secondary brain damage that are amenable to emergency maneuvers at the scene. In the literature, the influence of initial critical care management and early complications on the outcome of children with severe head injury is not clearly documented. METHODS AND RESULTS: We have prospectively examined the impact of the management at the scene on outcome in 40 children admitted to our Pediatric Intensive Care Unit with severe head injury (Glasgow Coma Scale < or = 8). The outcome of these children was assessed using the Glasgow Outcome Score (GOS). The results were evaluated by univariate and multivariate analysis. In our series the length of time before admission to an intensive care unit appears to have influenced the outcome among survivors, while the severity of injury is the only factor statistically associated with early complications (hypoxia and hypotension). With regard to the impact of early complications on outcome, the multivariate analysis showed that hypoxia and hypotension were significantly associated with GOS, independently of the GCS. CONCLUSION: Our results confirm the need for a precise treatment strategy for the initial management of children with neurotrauma to decrease the incidence of secondary brain damage attributable to hypoxia and hypotension. PMID- 11935246 TI - Intermittent vagal nerve stimulation in paediatric patients: 1-year follow-up. AB - OBJECT: Vagal nerve stimulation (VNS) has recently been proposed as a valid treatment for adult patients with drug-resistant partial epilepsy. Little experience in children has been reported. In order to evaluate the clinical efficacy and tolerance of VNS, we studied 13 paediatric patients with drug resistant partial epilepsy. METHODS: Improvement in seizure frequency was estimated by calculating the percentage of change in seizure frequency during each 3-month period following initiation of VNS, compared with the 3-month period prior to the implantation of the VNS device. The improvement in quality of life (QOL) was evaluated with the Vineland Behavior Adaptive Scale (VBAS). RESULTS: In all patients, the surgical procedure was well tolerated. A recent modification of the implantation technique needing only a single cervical incision, has further reduced the aesthetic damage, particularly in small children who have a reduced muscular mass. Three months after the surgical procedure, 10 of the 13 patients demonstrated a seizure reduction rate greater than 50%. At the 1-year follow-up these positive results were maintained: 6 out of 8 patients continued to demonstrate a seizure reduction rate greater than 50%. Comparison with the pre implantation period also showed a significant improvement in QOL in 4 out of 8 patients. We conclude that VNS is a valid treatment modality in children with drug-resistant partial epilepsy. PMID- 11935247 TI - Vertebral arterial dissection associated with Klippel-Feil syndrome in a child. AB - BACKGROUND: Vertebral artery dissection resulting in stroke is rare in children. We report here on a 7-year-old boy with Klippel-Feil abnormality, who presented with a pontine infarction after a supervised swimming session. METHODS: Evaluation after a second acute neurological event included a formal cerebral angiogram, which revealed a complete upper basilar artery occlusion and right vertebral arterial dissection. Cervical spine radiographs demonstrated an associated fusion of the C2 and C3 vertebrae. Anticoagulation therapy was initiated, and the neurological deficits associated with the pontine infarction resolved. Anticoagulation was discontinued after 6 months of therapy, with no recurrence of symptoms. CONCLUSION: Vertebral artery dissection may rarely be associated with Klippel-Feil abnormality in children. PMID- 11935248 TI - Anterior communicating artery aneurysm in a 3-year-old girl. AB - Intracranial aneurysms are rarely present in the paediatric age group, and such aneurysms within the anterior circulation are even more rare. Intracranial aneurysms in children differ from adult aneurysms in size, distribution, and incidence of symptoms. We report a 3-year-old girl with an anterior communicating artery aneurysm, who presented with subarachnoid haemorrhage. The patient underwent a pterional craniotomy and clipping of the aneurysm. The postoperative period was uneventful. We review the literature and discuss the characteristics of aneurysms in early childhood. PMID- 11935249 TI - Epidural hematoma from a cranial eosinophilic granuloma. AB - The formation of an epidural hematoma from an eosinophilic granuloma of the skull is an exceptional occurrence. A 9-year-old boy presented with severe headache, somnolence and vomiting following a minor head injury. Cranial computerized tomography scan showed a seemingly depressed skull fracture together with an epidural hematoma in evolution. A neoplasm and an epidural hematoma were removed at operation. Histopathological study of the excised mass confirmed the diagnosis of eosinophilic granuloma. PMID- 11935250 TI - Congenital disorder of glycosylation type Ia: benign clinical course in a new genetic variant. AB - The congenital disorders of glycosylation (CDG) are autosomal recessive disorders of N-glycans processing. Several different subtypes have been identified in recent years. Cerebellar atrophy is a characteristic finding in subtype Ia. We report clinical, imaging and genetic findings in a patient with a particularly benign clinical course, who had a normal CT at the age of 9 months and a new, previously undescribed, combination of mutations of the PMM gene locus on chromosome 16p13 (647,691). The 691 mutation has been described only in severe cases so far. This could indicate that genotype-phenotype correlation is lower than expected. PMID- 11935256 TI - Neuronal nitric oxide synthase (nNOS) immunoreactivity in the spinal cord of transgenic mice with G93A mutant SOD1 gene. AB - Immunohistochemical and quantitative analyses were used to examine the evolution of neuronal nitric oxide synthase (nNOS) with time in spinal motor neurons of transgenic mice with a G93A mutant Cu/Zn superoxide dismutase (SOD1) gene. Specimens from age-matched non-transgenic wild-type mice served as controls. In the controls, the anterior horn including the anterior horn neurons was not immunostained for nNOS. In the transgenic mice, at the age of 24 weeks (early presymptomatic), when no pathological change was observed in the spinal cord, anterior horn neurons were only occasionally immunostained for nNOS (0.3%). At the age of 28 weeks (late presymptomatic), nNOS-positive anterior horn neurons and their neuronal processes were occasionally observed (7.6%), and at the age of 32 weeks (early symptomatic), nNOS-positive anterior horn cells, including degenerated ones showing central chromatolysis, were frequently demonstrated (27.6%) and nNOS-positive cord-like swollen proximal axons were also observed in the anterior horns. nNOS expression in the anterior horn neurons was almost always observed in the somata. At the age of 35 weeks (end stage), neuronal loss of the anterior horn cells was severe, and nNOS-positive anterior horn neurons and cord-like swollen axons in the anterior horns were less prominent compared to those at the age of 32 weeks (33.8%), but many reactive astrocytes were immunostained for nNOS. Thus, nNOS immunoreactivity in the anterior horn neurons is observed as early as the presymptomatic stage and varies with the progression of the disease. The selective localization of positive nNOS immunoreactivity in the anterior horn neurons and degenerated ones in particular, and swollen proximal axons suggests that nNOS immunoreactivity may be involved in the degeneration of anterior horn neurons in this SOD1 transgenic mouse model. PMID- 11935257 TI - DL-Homocysteic acid application disrupts calcium homeostasis and induces degeneration of spinal motor neurons in vivo. AB - Excitotoxicity, autoimmunity and free radicals have been postulated to play a role in the pathomechanism of amyotrophic lateral sclerosis (ALS), the most frequent motor neuron disease. Altered calcium homeostasis has already been demonstrated in Cu/Zn superoxide dismutase transgenic animals, suggesting a role for free radicals in the pathogenesis of ALS, and in passive transfer experiments, modeling autoimmunity. These findings also suggested that yet confined pathogenic insults, associated with ALS, could trigger the disruption of calcium homeostasis of motor neurons. To test the possibility that excitotoxic processes may also be able to increase calcium in motor neurons, we applied the glutamate analogue DL-homocysteic acid to the spinal cord of rats in vivo and analyzed the calcium distribution of the motor neurons over a 24-h survival period by electron microscopy. Initially, an elevated cytoplasmic calcium level, with no morphological sign of degeneration, was noticed. Later, increasing calcium accumulation was seen in different cellular compartments with characteristic features of alteration at different survival times. This calcium accumulation in organelles was paralleled by their progressive degeneration, which culminated in cell death by the end of the observation time. These findings confirm that increased calcium also plays a role in excitotoxic lesion of motor neurons, in line with previous studies documenting the involvement of calcium ions in motor neuronal injury in other models of the disease as well as elevated calcium in biopsy samples from ALS patients. We suggest that intracellular calcium might be responsible for the interplay between the different pathogenic processes resulting in a uniform clinicopathological picture of the disease. PMID- 11935258 TI - Transneuronally altered dendritic processing of tangle-free neurons in Alzheimer's disease. AB - In Alzheimer's disease (AD), changes in dendritic morphology can be regarded as a result of an inherent disease-specific process associated with the formation of neurofibrillary tangles. Using three-dimensional morphometrical techniques and neuropatholologically staged tissue (Braak classification) of 32 cases, we demonstrate alterations in the dendritic length, branch order and number of segments of a tangle-free neuronal population in the AD-afflicted hippocampus, i.e. parvalbumin-containing cells of the fascia dentata. These alterations occurred primarily on the apical dendritic tree, the target of the entorhinal input. Mean of relative dendritic length, branch order and number of dendritic segments of apical dendrites decreased significantly, by 40-70% comparing stage V to stages 0 or I. In contrast, basal dendrites receiving no entorhinal input did not show significant changes. Entorhinal neurons projecting to the hippocampus are the first to be affected in AD and the first to die, resulting in hippocampal deafferentation. Therefore, this input-specific dendritic alteration of tangle free neurons suggests that AD is confounded with a transneuronal component resulting from deafferentation. Experiments showed that deafferentation results in altered dendritic geometry causing an impaired signal integration. Thus, transneuronally altered dendritic signal integration might occur in neurons devoid of the major intraneuronal hallmark of AD, i.e. the neurofibrillary tangle. PMID- 11935259 TI - Postischemic hyperthermia induces Alzheimer-like pathology in the rat brain. AB - This study addresses the effects of induced hyperthermia on post-ischemic rat brain evaluated histologically and/or immunohistochemically after 7-day, 2-month or 6-month survival. Hyperthermia (38.5 degrees - 40 degrees C) maintained (by heating the cage environment to 34-35 degrees C) for two consecutive periods of 5 and 9 h timed, respectively, from 4- and 21-h recirculation following 10-min global ischemia (two-vessel occlusion + hypotension) induced chronic neuronal death that became apparent in the rat forebrain from 7-day to 2-month survival. Associated immunohistochemical findings after 2 or 6 months of recovery included: (1) complement activation (membrane attack complex formation); (2) generalized overexpression of ubiquitin in surviving forebrain neurons; (3) persistent activation of macrophages; (4) presence of gemistocytic astrocytes in the hippocampus; (5) maturation of amyloid plaques (identified by immunohistochemistry using anti-human beta-A4 primary antibody) in cerebral cortex; and (6) intracellular deposits identified by anti-human hyperphosphorylated tau protein antibodies. This novel non-transgenic, self sustained model of neurodegeneration triggered by the association of two prevalent insults to the aging human brain (ischemia and hyperthermia) presents morphological features similar to those of Alzheimer's disease. This finding raises the possibility that febrile complications of acute brain injuries may similarly impair human cognitive function in the long run. PMID- 11935260 TI - Alterations of ubiquitin immunoreactivity in the hippocampal formation after perforant pathway lesion. AB - Immunohistochemical techniques were employed to examine the changes in free ubiquitin within the hippocampus 1, 3, 7, 14, and 30 days after a unilateral perforant pathway lesion occurred in the rat brain. Immunoreactivity for ubiquitin was remarkably decreased in the cell body and proximal dendrites of neurons throughout the hippocampus ipsilateral to the lesion at 1 day post lesion. At 3 days post-lesion, ubiquitin immunoreactivity was recovered in interneurons in the whole hippocampus as well as in mossy cells in the hilar region, although granule cells in the dentate gyrus and pyramidal cells in the CA1 subfield remained unlabeled, and pyramidal cells in the CA3 subfield demonstrated only weak immunoreactivity. In addition, we observed an increase in ubiquitin immunolabeling of the hilar neuropil ipsilateral to the lesion at 1 and 3 days post-lesion, and a decrease in immunolabeling in the inner portion of the molecular layer at 3 days post-lesion. All these alterations were transient, and by 7 days post-lesion, ubiquitin immunoreactivity was indistinguishable in the hippocampus ipsilateral to the lesion, compared to the controls. Immunoblot analysis also revealed a decrease in the amount of ubiquitin in the hippocampus ipsilateral to the lesion 1 and 3 days post-lesion. These data suggest that deafferentation of the perforant pathway results in transient reduction in free ubiquitin of the hippocampus, and that the ubiquitin system is involved in hippocampal plasticity following perforant lesions. PMID- 11935262 TI - Cerebellar cortical tau pathology in progressive supranuclear palsy and corticobasal degeneration. AB - Immunohistochemical localization of tau in the cerebellar cortex was carried out using a mouse monoclonal antibody against phosphorylation-dependent tau (AT8) in brain tissue (cerebellum) from 13 patients with progressive supranuclear palsy (PSP), 7 patients with corticobasal degeneration (CBD) and 5 age-matched control subjects. Purkinje cell somata that showed diffuse granular accumulation of cytoplasmic tau were found occasionally in 9 of the 13 patients with PSP (69%) and in 4 of the 7 patients with CBD (57%). Tau-positive doughnut-shaped structures were also found occasionally in the cerebellar molecular layer in 6 of the 13 patients with PSP (46%) and 2 of the 7 patients with CBD (29%). No tau immunoreactivity was detected in the cerebellar cortex in the control tissue. In the tissue from one patient with PSP, we also performed a double-labeling immunofluorescence study with anti-glial fibrillary acidic protein (GFAP) antibody and AT8, as well as an immuno-electron microscopic study with AT8. In tau-positive Purkinje cell somata and dendrites, the reaction product was localized mainly within the rough endoplasmic reticulum and free ribosomes. Tau positive doughnut-shaped structures were located in the GFAP-positive radial processes of Bergmann's glia and were present in the outer areas of inclusions reminiscent of Lewy bodies, which consist of aggregated pathological tau filaments. In conclusion, we have demonstrated a novel tau pathology that affects Purkinje cells and Bergmann's glia in patients with PSP and CBD, indicating that the cerebellar cortex can be involved in the disease processes in PSP and CBD. PMID- 11935261 TI - Chemokines are differentially expressed by astrocytes, microglia and inflammatory leukocytes in Toxoplasma encephalitis and critically regulated by interferon gamma. AB - The intracerebral formation of inflammatory infiltrates is a complex process, which may be regulated by chemokines. This study defines the kinetics and cellular sources of T cell- and macrophage-attracting chemokines in murine Toxoplasma encephalitis (TE) by ribonuclease protection assay, reverse transcription-PCR, in situ hybridization, and immunohistochemistry. Whereas astrocytes were the major source of interferon (IFN)-gamma-inducible protein-10 (CRG-2/IP-10) and monocyte chemoattractant protein (MCP)-1, microglia expressed RANTES, monokine induced by IFN-gamma (MuMIG) and occasionally CRG-2/IP-10 RNA. Despite being ubiquitously activated, only astrocytes and microglia confined to inflammatory infiltrates expressed chemokine genes. Intracerebral leukocytes transcribed RANTES, MuMIG, and occasionally CRG-2/IP-10 and MCP-1. IFN-gamma deficient mice failed to produce CRG-2/IP-10, MuMIG, RANTES and expressed macrophage inflammatory protein (MIP-1)alpha, MIP-1 beta, and MCP-1 mRNA at reduced levels, functionally resulting in a strongly reduced recruitment of leukocytes across the blood-brain barrier and prevented their further invasion of the brain parenchyma. Since T cells are the single source of IFN-gamma in TE, these findings indicate that T cells pave the way of leukocytes to parenchymatous parasites via IFN-gamma. PMID- 11935263 TI - Quantitative assessment of the innervation of epineurial arteries in the peripheral nerve by immunofluorescence: differences between controls and patients with peripheral arterial disease. AB - The peripheral nerve is supplied by the vasa nervorum. The epi- and perineurial vessels are innervated by an autonomic plexus, which plays a role in regulation of the endoneurial blood flow. This innervation is decreased in diabetes and alcohol polyneuropathy and seems to precede the development of diabetic polyneuropathy. A decreased innervation may therefore play a role in the development of polyneuropathy. In peripheral arterial disease (PAD) clinical and morphological features are present, related to severity of ischemia. To investigate the innervation of the vasa nervorum in severe ischemia, we performed immunofluorescence staining with the general neural marker protein gene product (PGP) 9.5 in whole mount preparations of epineurial vessels of nine sural nerves taken from patients with legs amputated because of severe PAD (59+/-15 years, mean +/- SD) and ten age-matched controls (61+/-24 years). In patients with PAD the nerve density of the perivascular plexus was decreased in comparison with controls (mean intercept density/mm +/- SD) 26.0+/-6.9 in PAD and 39.9+/-10.7 in controls, area% 6.0+/-1.6 in PAD and 9.9+/-2.6 in controls, both P<0.01, t-test). A decreased perivascular plexus may result in a diminished regulation of the endoneurial blood flow in patients with severe PAD. PMID- 11935264 TI - Degenerative and vascular lesions of the brain have synergistic effects in dementia of the elderly. AB - The relative importance of vascular and Alzheimer's disease (AD) lesions, their interaction in the development of cognitive impairment and the very existence of mixed dementia induced by the potentiation of both mechanisms remain controversial. The aim of this study was to assess whether the patients with infarcts and lacunes have fewer plaques and tangles than those without vascular lesions, for similar severity of clinical dementia. We performed a prospective clinicopathological study in elderly patients of a long-stay care unit. The severity of clinical dementia was assessed by psychometry performed according to standardized methods less than 6 months before death. A volumetric study of cerebral vascular lesions was performed at post-mortem study of the brain. The density of neuritic plaques (SP), Amyloid beta focal deposits (A beta FD), and neurofibrillary tangles (NFT) in the temporal and frontal isocortex was quantified. According to DSM III criteria, 28 of the 33 patients for whom autopsies were performed had dementia. Twenty-four of the included patients had degenerative or vascular lesions, or both. The volume of infarcts and lacunes was significantly correlated with the severity of cognitive impairment. The density of SP, A beta FD and NFT in the temporal and frontal isocortex was significantly lower when vascular lesions were present. For similar clinical severity of dementia, there were fewer AD lesions in patients with vascular lesions than in those without vascular lesions. PMID- 11935265 TI - Neuritic pathology is lacking in the entorhinal cortex, subiculum and hippocampus in middle-aged adults with schizophrenia, bipolar disorder or unipolar depression. AB - Earlier reports have provided conflicting results regarding the association between Alzheimer's disease (AD) and mental disorders. Using a well-characterized postmortem series of 40 middle-aged human brains, we have performed quantitative analysis of neurofibrillary tangles and senile plaques in the entorhinal cortex, subiculum and rostral hippocampus in 9 subjects with schizophrenia, 8 with bipolar disorder, 12 with depression, and 11 age- and sex-matched controls. No significant differences were found among the four groups. Our study indicates that the Alzheimer-type changes, which might be related to the likelihood of AD development later in life, are not increased in middle-aged subjects with mental illness. The result also supports the more recent reports that have demonstrated no increased incidence of AD in mentally ill patients. PMID- 11935266 TI - Patterns of protein nitration in dementia with Lewy bodies and striatonigral degeneration. AB - The synucleinopathies are a group of neurodegenerative disorders characterized by the presence of alpha-synuclein inclusions in neurons (Lewy body diseases, LBD) or glial cells (multiple system atrophies, MSA). Recently, nitration of alpha synuclein has been reported as the possible modification that induces its aggregation and deposition in these disorders. In this study we investigated the distribution and relationships of alpha-synuclein inclusions and 3-nitrotyrosine (3-NT), a marker of protein nitration through oxidative mechanisms, in brains diagnosed with LBD or MSA and control brains using double immunohistochemical techniques. In LBD cases, 3-NT colocalized with alpha-synuclein immunoreactivity in classic and cortical Lewy bodies and in dystrophic neurites in substantia nigra. However, most pale bodies and diffuse deposits in substantia nigra and Lewy neurites in hippocampus lack 3-NT immunoreactivity. A majority of cases showed diffuse cytoplasmic 3-NT staining in pyramidal cells of the CA2-3 regions of the hippocampus that was independent of alpha-synuclein deposits. All MSA cases showed 3-NT immunoreactivity in glial inclusions. 3-NT neuronal staining was restricted to pontine nuclei with three cases showing nuclear and one case cytoplasmic staining. There was no colocalization of 3-NT nuclear immunoreactivity with alpha-synuclein-immunopositive nuclear inclusions in pontine neurons. These data show that protein nitration in LBD and MSA cases has a widespread distribution and is not only associated with the alpha-synuclein deposits. The presence of alpha-synuclein-positive deposits lacking 3-NT immunoreactivity suggests that nitration is not a prerequisite for alpha synuclein deposition. PMID- 11935267 TI - Altered trafficking and adhesion function of MPZ mutations and phenotypes of Charcot-Marie-Tooth disease 1B. AB - We previously reported familial cases characterized by Charcot-Marie-Tooth disease (CMT) phenotype with abnormal myelin foldings and MPZ Ile62Phe mutation. To further clarify the molecular mechanisms in this family, we produced wild-type MPZ, Ile62Phe mutant and other mutations in the neighboring regions producing thin myelin sheaths (Ser63del, Ser63Cys and Ser63Phe) by site-specific mutagenesis and transfected these into rat pheochromocytoma cells (PC12). We investigated the expression and aggregation properties of the MPZ protein through immunoblotting, immunohistochemical staining and adhesion assay. MPZ protein with Ile62Phe mutation was immunohistochemically detectable mainly in the plasma membrane of the cells, and it induced a cell aggregation behavior different from the other mutations or the wild-type MPZ. These studies suggested that MPZ Ile62Phe mutation in CMT1B with abnormal myelin folding induced dysregulation of adhesion function of the MPZ protein in a manner unlike those seen in cells with other mutations. The present study provides evidence that the site and nature of amino acid substitutions in the MPZ protein are closely related to the abnormal myelination in CMT1B. PMID- 11935268 TI - Immunohistochemical analysis of anti-Hu-associated paraneoplastic encephalomyelitis. AB - The precise immune mechanisms of neuronal death in anti-Hu-associated paraneoplastic encephalomyelitis (PEM) are unclear. We performed an immunohistochemical study on postmortem brain tissue from 11 patients with anti Hu-associated PEM to further characterize the immune reaction and to ascertain possible mechanisms of neuronal death. To analyze inflammatory infiltrates, antibodies against lymphocyte subpopulations (CD3, CD20, CD4, CD8), macrophage and activated microglia (CD68), major histocompatibility complex (MHC) classes I and II (HLA-ABC and HLA-DR), and the intercellular adhesion molecules (ICAM) -1 and -3 were used. Cell death mechanisms were defined using antibodies against the cytotoxic protein TIA-1, the C9neo component of complement, the Fas receptor (CD95) and its ligand, the apoptosis effector activated caspase-3, and the apoptosis inhibitor Bcl-2. A great number of T cells expressing the cytotoxic protein TIA-1 was observed, mainly in clusters around neurons. ICAM-1 immunoreactivity was increased in the neuropil and reactive astrocytes in areas of inflammation within the central nervous system and in satellite cells of pathological dorsal root ganglia surrounding apparently normal sensory neurons. By contrast, Fas, FasL, C9neo, and activated caspase-3 immunoreactivities were negative in pathological areas. Bcl-2 immunoreactivity was found in satellite cells, but not in sensory neurons of normal and pathological dorsal root ganglia. Our data point out to an induction of a cytotoxic, non-apoptotic, neuronal death in anti-Hu-associated PEM. The increased ICAM-1 immunoreactivity may favor the infiltration of lymphocytes in the pathological areas. PMID- 11935269 TI - Alpha-synuclein-immunoreactive deposits in human and animal prion diseases. AB - Prion related disorders are associated with the accumulation of a misfolded isoform (PrPsc) of the host-encoded prion protein, PrP. There is strong evidence for the involvement of unidentified co-factors in the PrP to PrPsc conversion process. In this study, we show alpha-synuclein-immunoreactive deposits in the central nervous system of various prion diseases (sporadic, iatrogenic and new variant Creutzfeldt-Jakob diseases, and experimental scrapie of hamsters). alpha Synuclein accumulated close to PrPsc deposits but we did not observe strict colocalization of prion protein and alpha-synuclein immunoreactivities particularly in PrPsc plaques. alpha-Synuclein is thought to be a key player in some neurodegenerative disorders, is able to interact with amyloid structures and has known chaperone-like activities. Our results, in various prion diseases, suggest a role for alpha-synuclein in regulating PrPsc formation. PMID- 11935270 TI - Motor neuron disease with dementia combined with degeneration of striatonigral and pallidoluysian systems. AB - This study concerns an autopsy case of motor neuron disease with dementia (MND-D) that exhibited unusual clinical and neuropathological findings. The patient was a Japanese man without any relevant family history who was 60 years old at the time of death. His clinical manifestation included character change at the age of 54, followed by frozen gait, dysarthria and bradykinesia and he was diagnosed with Parkinson's disease. He gradually developed spastic paresis and died of respiratory failure 6 years after onset of the illness. Neuropathological examinations showed prominent degeneration in the striatonigral and pallidoluysian systems in addition to the neuronal loss and microvacuolation in the second to third layers of the frontal and temporal cortex, the involvement of the upper and lower motor neuron systems and the presence of ubiquitinated neuronal inclusions. To our knowledge, five cases of motor neuron disease (MND) combined with pallido-nigro-luysian atrophy (PNLA) have been reported previously, but the present case is the first report of MND-D combined with the degeneration of the striatonigral and pallidoluysian systems. Such an association may represent more than a coincidental occurrence, and it suggests that MND-D is not simply a disease of the motor neuron system but a multisystem degeneration. PMID- 11935271 TI - Co-occurrence of Parkinson's disease with progressive supranuclear palsy. AB - Parkinson's disease (PD) and progressive supranuclear palsy (PSP) are distinct neurodegenerative disorders. We describe an 81-year-old woman with 3 years of progressive gait unsteadiness, frequent falls, and mild cognitive dysfunction, all considered clinically to be an early fronto-temporal neurodegenerative disorder. She died of an acute myocardial infarction. Examination of her brain revealed alpha-synuclein- and tau-positive inclusions diagnostic of PD and PSP. Immunoelectron microscopy and Western blot analysis confirmed combined PD/PSP. This case provides strategies for the reliable molecular validation of concomitant PD and PSP, and demonstrates the utility of these techniques in patients with atypical clinical presentations. PMID- 11935272 TI - High prevalence of resistance to APC in young patients with retinal vein occlusion. AB - Resistance to activated protein C (APC) is among the coagulation disorders that have been implicated in retinal vein occlusion. However, since retinal vascular occlusions may be due to a combination of several mechanisms, the question of whether thrombophilic anomalies are pathogenic for this disorder remains controversial. In the current study, we investigated the prevalence of APC resistance in patients with retinal vein occlusion with reference to age and various cardiovascular risk factors. A cohort of 142 consecutive patients with retinal vein occlusion and a control group of 128 subjects matched for age, sex and several risk factors were screened for resistance to APC. Both cohorts were divided into two subgroups, according to the patient's age (< or =45 or >45 years) at the time of the retinal vein occlusion or a previous thromboembolic event. The proportion of individuals with resistance to APC was higher in the patient group (13 of 142; 9.1%) when compared to controls (6 of 128; 4.7%). Moreover, patient age < or =45 years by the time of the retinal vein occlusion or a previous thromboembolic event was significantly associated with a high prevalence of APC resistance (17%). By contrast, resistance to APC was present in 5 of 95 cases (5.3%) in the patient group >45 years and in 4 of 83 (4.8%) young controls. Our results indicate that APC resistance is highly prevalent in patients with retinal vein occlusion at age < or =45 years and/or with a history of thrombosis at this age. By contrast, the prevalence of APC resistance in patients who suffered a retinal vein occlusion when they were older than 45 years and had no history of thromboembolism appears to be similar to that seen in healthy control subjects or in the normal population. Selective screening may be helpful in identifying retinal vein occlusion patients with thrombophilic defects. PMID- 11935274 TI - Glaucoma secondary to pars plana lensectomy for congenital cataract. AB - BACKGROUND: To identify risk factors related to the onset of secondary glaucoma in eyes that have undergone pars plana lensectomy for congenital cataract. METHODS: We investigated intraocular pressure levels after pars plana lensectomy for congenital cataract in 80 eyes of 44 patients who were followed up for at least 36 months. Mean age at the time of pars plana lensectomy was 11+/-18 months. Mean follow-up period was 116+/-57 months. RESULTS: An elevation in intraocular pressure (>20 mmHg) was found in 21 eyes of 12 patients (26%). In 7 of these 21 eyes surgical treatment was required. The incidence of the coexistence of microcornea (or microphthalmos) with congenital cataract was significantly higher in the glaucoma group than that in nonglaucoma group. The final visual acuity in the nonglaucoma group was significantly better than that in the glaucoma group. CONCLUSION: Our results show the importance of careful follow-up of intraocular pressure after pars plana lensectomy for congenital cataract, especially when microcornea (or microphthalmos) is present. PMID- 11935273 TI - Angiographic findings in patients with exudative age-related macular degeneration. AB - PURPOSE: Exudative age-related macular degeneration (ARMD) is the leading cause for severe visual loss in the elderly. A series of clinical trials has established the efficacy of laser photocoagulation and photodynamic therapy (PDT) in special subgroups of patients with exudative ARMD. We analyzed the fluorescein and indocyanine green angiograms (ICG) of patients with exudative ARMD to characterize the nature of the CNV in order to estimate the percentage of patients amenable for one of the therapies. METHODS: Patients with acute symptoms related to exudative ARMD were recruited for the study. All patients underwent fluorescein and ICG angiography. We analyzed the type of the CNV (classic, occult, or mixed) and location relative to the fovea from the fluorescein angiograms. Additionally, we recorded the presence of a visible capillary network in the early ICG angiograms, as well as hot spots and plaque hyperfluorescence in the late ICG angiographic frames. RESULTS: We included 191 patients aged 53-92 years in the study. The angiograms revealed in 17 (9%) patients classic CNV (12 subfoveal), in 114 (60%) occult CNV, and in 60 (31%) mixed CNV. Of these 19 had a classic component >50%. In the early ICG angiograms a capillary network was visible in 50 eyes with occult CNV. Hot spots were present in 29 eyes and a hyperfluorescent plaque in 54 eyes. CONCLUSION: In total, only 18% of eyes included in our study were eligible for laser photocoagulation or for photodynamic therapy (PDT). These numbers highlight the deficiencies of current treatment modalities for exudative ARMD. For this reason, alternative treatment should be further investigated to offer better treatment to patients with exudative ARMD. PMID- 11935275 TI - Hematologic abnormalities associated with various types of retinal vein occlusion. AB - BACKGROUND: The objective of this study was two-fold: (1) to investigate hematologic abnormalities associated with various types of retinal vein occlusion (RVO) and comparison of their prevalence among those various types of RVO; (2) to review the conflicting literature on the subject, to place the information in perspective. METHODS: In patients with various types of RVO seen in our clinic since 1973, we conducted planned prospective studies on the prevalence of: (1) routine hematologic tests (535 patients) and (2) certain special hematologic parameters (platelet aggregation, antithrombin III, and proportional, variant(2) globulin in 110, 81 and 91 patients, respectively). Patients were categorized into six types of RVO, based on defined criteria: non-ischemic and ischemic central RVO (CRVO), non-ischemic and ischemic hemi-CRVO (HCRVO), and major and macular branch RVO (BRVO). The patients had a detailed ophthalmic, systemic and hematologic evaluation. The data were abstracted and analyzed retrospectively from the detailed information originally collected prospectively in the patients' records. For data analysis, patients were divided into young, middle-aged and elderly. Observed prevalence rates of hematologic abnormalities were estimated. Logistic regression, adjusting for age and gender, was used to compare the observed prevalence of hematologic abnormalities among the various types of RVO. RESULTS: No generalizations about the prevalence of hematologic disorders in all six types of RVO are possible. Ischemic CRVO showed a significantly higher prevalence of abnormal hematocrit ( P=0.044), hemoglobin ( P=0.018), and blood urea nitrogen ( P=0.025) than non-ischemic CRVO, while a significantly higher prevalence of abnormal antinuclear antibody (ANA; P=0.049) was seen in non ischemic CRVO than in ischemic CRVO. There was a significant ( P=0.011) difference in the prevalence of abnormal uric acid among the three main RVO groups (CRVO, HCRVO, BRVO), highest in BRVO and lowest in HCRVO. There was a higher prevalence of abnormal glucose ( P=0.069) and ANA ( P=0.071) in CRVO+HCRVO than in BRVO. Results of special hematologic studies are given. CONCLUSIONS: Our study showed that a variety of hematologic abnormalities may be seen in association with different types of RVO, and any generalization about these disorders applied to all RVO patients may be misleading. The evidence of our study and in the literature indicates that there is no good reason why all patients with RVO should be subjected to extensive, expensive, special hematologic and hypercoagulability investigations, unless, of course, there is some clear indication; the routine, inexpensive hematologic evaluation is usually sufficient for RVO patients. Treatment with anticoagulants or platelet anti aggregating agents may adversely influence the visual outcome, without any evidence of protective or beneficial effect. PMID- 11935276 TI - Effect of pars plana vitrectomy for proliferative diabetic vitreoretinopathy on preexisting diabetic maculopathy. AB - BACKGROUND: Vitrectomy is the treatment of choice for proliferative diabetic vitreoretinopathy with tractions and persistent vitreous hemorrhage. Since vitrectomy has recently been discussed as a possible therapy for diabetic maculopathy as well, we were especially interested in studying the change in diabetic maculopathy following surgery. For that purpose a grading system developed at our clinic was used. METHODS: In a retrospective study we evaluated fundus photographs and fluorescein angiograms of 33 eyes of 30 patients who had undergone vitrectomy for proliferative diabetic vitreoretinopathy at our clinic between 1990 and 1997. In all eyes diabetic maculopathy was present at the time of surgery. The grading was performed using preoperative images and images taken a median of 18 months postoperatively. RESULTS: Best corrected visual acuity increased by 3.7 lines on average. Intraretinal dot and spot hemorrhages, hard exudates, microaneurysms on fundus photos, and leakage and cysts on fluorescein angiograms decreased after surgery. The extent of the foveolar avascular zone and the extent of the perifoveolar ischemic area worsened, however. CONCLUSION: Vitrectomy seems to help diabetic eyes not only by removal of membranes, tractions, and vitreous hemorrhage; it seems to have a positive influence on the course of diabetic maculopathy as well. We suspect that the removal of the posterior vitreous membrane is one of the crucial factors in interrupting the disease process. From these results the indication for vitrectomy in diabetic patients may have to be reconsidered and extended to include diabetic maculopathy prior to the development of ischemia. PMID- 11935277 TI - Multifocal electroretinogram: age-related changes for different luminance levels. AB - BACKGROUND: Age-related changes in the first-order multifocal electroretinogram (mfERG) responses were measured for two different luminance levels (200 and 700 cd.m(-2)). The relative contribution of optical and neural factors to senescent change in response was evaluated. METHODS: Data were obtained from one eye of each of 71 normal phakic subjects, age 9-80 years. The mfERG responses were recorded with the 7" stimulus-refractor unit (EDI) and VERIS 4.3 using the following protocol: bipolar contact lens, 103 hexagons, consecutive stimulation with 200 and 700 cd.m(-2), pupils > or =6 mm, amplification of 10(5), filter cut offs at 10 and 300 Hz. RESULTS: Age-correlated decreases in amplitude and response density and increases in P1 implicit time were found for both luminance levels. The mean response density (nV.deg(-2)) was higher for the 700 cd.m(-2) stimulus, but the rate of change with age was not significantly different from that obtained with the 200 cd.m(-2) stimulus. Implicit time was not significantly different for the two light levels, nor was the rate of change with age. The decrease in response density and the increase in implicit time with age were significant across all retinal regions, dividing the 50 deg stimulus into six concentric rings. Age-related change in response density was greatest for the central retina and decreased with increasing retinal eccentricity. CONCLUSION: Log mfERG response changes linearly as a function of age. Analyses of the effects of reduced ocular media transmission and increased stray light, along with ancillary data obtained from pseudophakes, imply that age-related changes in the mfERG are due to both optical and neural factors. PMID- 11935278 TI - Morphological and functional damage of the retina caused by intravitreous indocyanine green in rat eyes. AB - BACKGROUND: This study was designed to investigate the influence of intravitreal indocyanine green (ICG) on retinal morphology and function. METHODS: Brown Norway rats eyes ( n=24) were vitrectomized by the injection of 0.05 ml of 100% SF(6) gas. Two weeks later, ICG solution was injected into the vitreous cavity of vitrectomized eyes at a dose of 25 mg/ml, 2.5 mg/ml, 0.25 mg/ml or 0.025 mg/ml (0.05 ml/eye). Retinal toxicity was histologically assessed by light microscopy on day 10. The retinal function was also evaluated by electroretinography (ERG) in the low-dose groups (0.25 mg/ml and 0.025 mg/ml) after 10 days and again after 2 months,. Sham-operated eyes (SF(6) injected followed by 0.05 ml of BSS plus, n=6) were used as controls. RESULTS: In the high-dose group (25 mg/ml ICG), the retinal structure was severely deformed and the retinal pigment epithelium partly disappeared. In eyes with 2.5 mg/ml ICG, the retinal structure was also affected but less strongly so than with 25 mg/ml. No apparent pathologic change was observed in the low-dose groups (0.25 mg/ml or 0.025 mg/ml) by light microscopy. In contrast, 10 days later the amplitude of dark-adapted a- and b-waves of ERGs in the eyes of low-dose group rats were found to have decreased. In addition the light-adapted b-waves did not change significantly. These changes remained for 2 months. CONCLUSION: Even at a low dose (0.025 mg/ml), intravitreous ICG induced functional damage of the retina without any apparent morphological damage. This information should be taken into account when clinically administering ICG into the vitreous cavity. PMID- 11935279 TI - Caspase-3 inhibitor transiently delays inherited retinal degeneration in C3H mice carrying the rd gene. AB - BACKGROUND: The effect of a caspase-3 inhibitor on retinal degeneration in C3H mice carrying the rd gene, a mutation of a rod-specific phosphodiesterase, was investigated. METHODS: A quantity of 2 mg/kg of Ac-DEVD-CHO, as inhibitor, was injected intraperitoneally every other day from 8 days of age, and retinal damage was compared with that in saline-treated C3H mice at 13 days (1 day after the third treatment) and 17 days of age (1 day after the fifth treatment). Retina of ICR mice not carrying rd gene was also evaluated under the same protocol. The efficacy of Ac-DEVD-CHO was evaluated based on total retinal thickness and outer retinal thickness (thickness of outer nuclear layer and photoreceptor layer). An apoptotic index and a cell proliferation index for the photoreceptor cells, at 13 days of age, were calculated based on terminal deoxynucleotidyl transferase mediated deoxyuridine triphosphate-digoxigenin nick-end labeling (TUNEL) and proliferating cell nuclear antigen (PCNA) labeling, respectively. RESULTS: At 13 days of age, total and outer retinal thickness in saline-treated C3H mice were 140.3 microm and 37.5 microm, compared with 160.4 microm and 49.5 microm, respectively, in Ac-DEVD-CHO-treated C3H mice ( P<0.01, respectively). In ICR mice, total and outer retinal thickness were 182.1 microm and 90.9 microm, respectively, in saline-treated mice and 183.8 microm and 89.6 microm in Ac-DEVD CHO-treated mice (not significant). At this time, the TUNEL index was 23.52 cells/10(4) microm (2) of outer nuclear layer in saline-treated C3H mice; Ac-DEVD CHO treatment significantly reduced this value to 18.73 cells/10(4) microm(2) ( P<0.05). The TUNEL index in saline- and Ac-DEVD-CHO-treated ICR mice was 0.59 cells/10(4) microm(2) and 0.80 cells/10(4) microm(2), respectively (not significant); Ac-DEVD-CHO treatment had no influence on normally developing retina. The PCNA index was not affected by Ac-DEVD-CHO-treatment. However, at 17 days of age, Ac-DEVD-CHO treatment did not ameliorate retinal degeneration. CONCLUSIONS: The caspase-3 inhibitor was transiently effective in delaying retinal degeneration through inhibition of the apoptosis of photoreceptor cells in rd gene-carrying mice. The use of caspase-3 inhibitors may have therapeutic applications in the treatment of human retinal degeneration. PMID- 11935280 TI - Morphological evidence of M cells in healthy canine conjunctiva-associated lymphoid tissue. AB - PURPOSE: To characterize the follicle-associated epithelium (FAE) and organized lymphoid nodules from dog nictitating membranes to determine if canine conjunctiva-associated lymphoid tissue (CALT) contains M cells analogous to those described in other regions of mucosa-associated lymphoid tissue (MALT). METHODS: Nictitan lymphoid follicles from 15 healthy dogs (30 eyes) were harvested immediately post-mortem. Twelve follicles from each nictitating membrane were isolated. Four follicles from each eye of 10 dogs were examined by light microscopy, transmission electron microscopy and scanning electron microscopy. Five of the 10 dogs were treated with a heat-killed staphylococcal topical suspension immediately prior to euthanasia. Nictitan follicles from five other dogs were processed for immunohistochemistry to characterize follicle lymphocyte populations. RESULTS: The FAE overlying CALT demonstrated morphology characteristic of M cells, including attenuated apical cell surface with blunted microvilli and microfolds, invaginated basolateral membrane forming a cytoplasmic pocket containing lymphocytes and macrophages, and diminished distance between the apical and pocket membrane. Heat-killed bacteria were bound to the surface and transcytosed to the cytoplasmic pocket of CALT M cells. Immunohistochemistry of organized lymphoid tissue subtending the FAE demonstrated B-cell germinal centers with T-cell predominant apical caps. CONCLUSIONS: In canine CALT, the FAE overlying lymphoid follicles, as well as the distribution of T and B lymphocytes subtending this region, contain morphologic and functional features analogous to MALT described in other regions. Documentation of canine conjunctival M cells is of clinical relevance in the study of primary ocular diseases, as well as a potential means of vaccination or drug delivery. PMID- 11935281 TI - Octreotide inhibits growth factor-induced bovine choriocapillary endothelial cells in vitro. AB - BACKGROUND: Response to growth factors by the choriocapillaris is one of the main aspects in the development and progression of choroidal neovascularization in age related macular degeneration (AMD). The aim of the study was to examine the effect of octreotide--an analogue of somatostatin with possible favorable effects in the treatment of age-related macular degeneration--on growth factor-activated bovine choriocapillary endothelial cells (BCEC). METHODS: Cultivated BCEC were stimulated with 20 ng/ml VEGF, 5 ng/ml bFGF, 50 ng/ml TGFbeta(2), or 50 ng/ml IGF I for 24 h. Afterwards, the cells were incubated for a further 24 h with octreotide (10(-6) M). BCEC proliferation was determined by [(3)H]-thymidine incorporation. The experiments were performed in normal conditions and in O(2) limited (5% O(2)) atmosphere. RESULTS: VEGF, bFGF, and IGF-I significantly stimulated BCEC proliferation under normoxic as well as under hypoxic conditions ( P<0.05); the value for TGFbeta(2)stimulation was not significant. Under normoxic conditions, after addition of octreotide to VEGF-, bFGF-, and IGF-I stimulated cells there was no longer any significant [(3)H]-thymidine incorporation compared to the control medium. Under hypoxic conditions similar results were obtained after addition of octreotide to VEGF-, bFGF-, and IGF-I stimulated cells. CONCLUSION: Octreotide has inhibitory properties after activation of BCEC with growth factors under normoxic and hypoxic conditions in vitro. Since activation of these cells is a crucial event in the development and progression of AMD, octreotide may be a potential drug for treatment of choroidal neovascularization in these patients. PMID- 11935282 TI - Long-term effects of short-term retinal bleb detachments in rabbits. AB - PURPOSE: To examine the effects of saline-induced bleb detachments in rabbit retina. METHODS: Retinal bleb detachments were produced by the injection of 50 microl of balanced salt solution (BSS) into the subretinal space of one eye of each of six rabbits using a glass pipette with a flat tip, 50 microm in diameter. The retina was examined by biomicroscopy, scanning laser ophthalmoscopy (SLO), auto-fluorescence and simultaneous fluorescein and indocyanine green (ICG) angiography. Histological examination was carried out at 1, 2, 3 and 4 months after surgery. RESULTS: All rabbits showed leakage of fluorescein for at least a day after detachment, but within 1 month the leakage ceased. ICG staining developed gradually at the level of the RPE or Bruch's membrane near sites of previous staining. Lipofuscin fluorescence also developed gradually around areas of staining. Histology revealed the source of the excessive lipofuscin to be in the RPE layer, especially in cells migrating away from Bruch's membrane. CONCLUSIONS: Short-term bleb detachments cause a transient breakdown in the blood retinal barrier, long-term ICG staining at or deep to the RPE layer, hyperlipofuscinosis and migration of the RPE. The abnormal lipofuscin accumulation is apparent on fluorescence ophthalmoscopy and can be confused with markers such as green fluorescent protein. PMID- 11935285 TI - Endosomes: multipurpose designs for integrating housekeeping and specialized tasks. AB - In most cells the endocytic system is organized following a common concept that allows for the integrative handling of a variety of housekeeping functions. In addition, variations on the general scheme provide for specialized endosome-based pathways that occur only in specific cell types. The diversity of endosomal functions is not only reflected by characteristic molecular compositions, but also mirrored in their morphological organization. In this review we will first describe the general outline of the endocytic system by combining kinetic, morphological, molecular, and functional definitions. In the second part, adaptations of endosomes that allow their functioning in specialized processes, such as antigen presentation, synaptic vesicle formation, and glucose transport, will be addressed. PMID- 11935286 TI - Vesicle formation and trafficking in endothelial cells and regulation of endothelial barrier function. AB - Endothelial barrier function is regulated in part by the transcellular transport of albumin and other macromolecules via endothelial caveolae (i.e., this process is defined as transcytosis). Using pulmonary microvascular endothelial cells, we have identified the specific interactions between a cell surface albumin-docking protein gp60 and caveolin-1 as well as components of the signaling machinery, heterotrimeric G protein (G(i))- and Src-family tyrosine kinase. Ligation of gp60 on the apical membrane induces the release of caveolae from the apical membrane and activation of endocytosis. The formed vesicles contain the gp60-bound albumin and also albumin and other solutes present in the fluid phase. Vesicles are transported in a polarized manner to the basolateral membrane, releasing their contents by exocytosis into the subendothelial space. The signaling functions of G(i) and Src are important in the release of caveolae from the plasma membrane. The Src-induced phosphorylation of caveolin-1 is crucial in regulating interactions of caveolin-1 with other components of the signaling machinery such as G(i), and key signaling entry of caveolae into the cytoplasm and endocytosis of albumin and other solutes. This review addresses the basis of transcytosis in endothelial cells, its central role as a determinant of endothelial barrier function, and signaling mechanisms involved in regulating fission of caveolae and trafficking of the formed vesicles from the luminal to abluminal side of the endothelial barrier. PMID- 11935287 TI - Biogenesis and exocytosis of Weibel-Palade bodies. AB - Vascular endothelial cells contain typical, elongated vesicles, the so-called Weibel-Palade bodies, which serve as a storage compartment for von Willebrand factor (VWF), a plasma protein that plays an essential role in controlling the adhesion and aggregation of platelets at sites of vascular injury. Upon activation of endothelial cells by agonists such as thrombin, epinephrine or histamine, the Weibel-Palade bodies fuse with the plasma membrane and release their contents into the blood circulation. This process provides an adequate means by which endothelial cells can actively participate in controlling the arrest of bleeding upon vascular damage. Besides VWF, Weibel-Palade bodies contain a subset of other proteins, including interleukin-8 (IL-8), P-selectin and endothelin. Similar to VWF, these proteins are transported to the outside of the cell upon stimulation and may control local or systemic biological effects, including inflammatory and vasoactive responses. Apparently, endothelial cells are able to create a storage pool for a variety of bioactive molecules which can be mobilised upon demand. Endothelial cells that are deficient of VWF synthesis are not only unable to form Weibel-Palade bodies, but also lack the ability to store IL-8 or P-selectin or release these proteins in a regulated manner. It thus appears that VWF not only plays a prominent role in controlling primary haemostasis, but also may modulate inflammatory processes through its ability to target inflammatory mediators to the regulated secretion pathway of the endothelium. PMID- 11935288 TI - In vivo and in vitro processing of recombinant pro-von Willebrand factor. AB - Von Willebrand factor (vWF) is synthesized in endothelial cells as pre-pro-vWF and processed intracellularly to propeptide (vWFpp) and mature vWF. Recombinant pro-vWF when infused into animals can also be processed extracellularly in vivo. Within 1 h of infusion in a dog and mice the multimer pattern changed to that typically seen in mature vWF indicating that propeptide cleavage from unprocessed vWF occurs extracellularly in the circulation. Incubation of a recombinant pro vWF preparation with canine and human vWF-deficient plasma induced a time dependent decrease in pro-vWF antigen and an increase in vWFpp antigen without changing total vWF antigen or collagen-binding activity. Multimer analysis showed the gradual transformation of the pro-vWF multimers to mature vWF multimers and cleaved vWFpp was visualized on autoradiograms of SDS-polyacrylamide electrophoresis gels using (125)I-labeled pro-vWF. When recombinant pro-vWF was incubated with increasing amounts of purified thrombin, the extent of pro-vWF processing was dose dependent. The specific cleavage of vWFpp was confirmed by immunoblots using an anti-vWFpp antibody and by amino-terminal amino acid analysis. Hirudin preconditioning of vWF-deficient mice attenuated processing of infused recombinant pro-vWF suggesting that thrombin plays a part in the processing events in vivo. PMID- 11935289 TI - Pathways followed by ricin and Shiga toxin into cells. AB - The plant toxin ricin and the bacterial toxin Shiga toxin belong to a group of protein toxins that inhibit protein synthesis in cells enzymatically after entry into the cytosol. Ricin and Shiga toxin, which both have an enzymatically active moiety that inactivates ribosomes and a moiety that binds to cell surface receptors, enter the cytosol after binding to the cell surface, endocytosis by different mechanisms, and retrograde transport to the Golgi apparatus and the endoplasmic reticulum (ER). The toxins can be used to investigate the various transport steps involved, both the endocytic mechanisms as well as pathways for retrograde transport to the ER. Recent studies show that not only do several endocytic mechanisms exist in the same cell, but they are not equally sensitive to removal of cholesterol. New data have revealed that there is also more than one pathway leading from endosomes to the Golgi apparatus and retrogradely from the Golgi to the ER. Trafficking of protein toxins along these pathways will be discussed in the present article. PMID- 11935290 TI - Golgi apparatus and TGN during endocytosis. AB - Wheat germ agglutinin labelled with horseradish peroxidase (WGA) was used for analyses of endosomal compartments and Golgi apparatus in HepG(2) hepatoma cells during early and late periods of endocytosis. WGA was rapidly transferred into the Golgi region. Transport of internalised WGA into the Golgi apparatus could be classified in three stages. A short stage I, characterised by predominance of vesicular endosomes, was followed by stage II showing new formations of extended endocytic trans Golgi networks (TGNs); the endocytic TGNs comprised reticular and globular parts, showed intimate associations with segments of the endoplasmic reticulum and budding of multiple coated vesicles. Parts of the endocytic TGNs associated with trans Golgi cisternae and became integrated into Golgi stacks. During stage III, concomitantly with integration into the stacks, the endocytic TGNs decreased in size and stacked Golgi cisternae became prominent endocytic compartments. Our results show that endocytosis of WGA is connected with extensive membrane dynamics at the trans Golgi side: an endocytic TGN is newly formed, increases in size and is consumed again. The findings suggest that incorporation of TGN elements into Golgi stacks provides a mechanism for uptake of internalised WGA into the Golgi apparatus. PMID- 11935291 TI - Formation, isomerisation and reduction of disulphide bonds during protein quality control in the endoplasmic reticulum. AB - Efficient protein folding and quality control in the endoplasmic reticulum (ER) require that disulphide bonds are formed in nascent proteins, isomerised during assisted folding and reduced in terminally misfolded molecules. Recent findings in yeast and mammalian cells indicate that specific protein-protein interactions underlie redox control in the ER, allowing these competing reactions to occur simultaneously during protein quality control. PMID- 11935292 TI - The importance of trimming reactions on asparagine-linked oligosaccharides for protein quality control. AB - Although the biochemistry of early trimming reactions by glucosidases and ER mannosidases occurring on asparagine-linked oligosaccharides has been known for a long time, their involvement in quality control of protein folding has become apparent only more recently. Here we review the evidence for the involvement of specific oligosaccharide trimming intermediates such as Glc(1)Man(9)GlcNAc(2) and Man(8)GlcNAc(2) B isomer in this fundamental cellular process and the subcellular distribution of components of the protein quality control machinery which indicates the involvement of both the ER and pre-Golgi intermediates in this process. In addition, recent studies on the subcellular distribution of endomannosidase in conjunction with previously obtained biochemical data will be reviewed which demonstrate that an alternative deglucosylation pathway exists in pre-Golgi intermediates and the Golgi apparatus. PMID- 11935293 TI - The biological role of the Alzheimer amyloid precursor protein in epithelial cells. AB - Proteolytic processing of the Alzheimer amyloid precursor protein (APP) results in the generation of at least two distinct classes of biologically relevant peptides: (1) the amyloid beta peptides which are believed to be involved in the pathogenesis of Alzheimer's disease and (2) the soluble N-terminal ectodomain (sAPP) which exhibits a protective but as yet ill-defined effect on neurons and epithelial cells. In this report we present an overview on the functions of sAPP as an epithelial growth factor. This function involves specific binding of sAPP to membrane rafts and results in signal transduction and various physiological effects in epithelial cells as different as keratinocytes and thyrocytes. At nanomolar concentrations sAPP induces a two to fourfold increase in the rate of cell proliferation and cell migration. Specific inhibition of APP expression by antisense techniques results in decreased sAPP release and in reduced proliferative and motogenic activities. Proliferation and migration are known to be part of complex processes such as wound healing which, therefore, might be facilitated by the growth factor function of sAPP. PMID- 11935294 TI - Upregulation of the expression of endogenous Mdr1 P-glycoprotein enhances lipid translocation in MDCK cells transfected with human MRP2. AB - Various ABC transporters can translocate lipid molecules from the cytoplasmic into the exoplasmic leaflet of the plasma membrane bilayer. Two of these, MDR1 P glycoprotein (Pgp) and MRP1, are multidrug transporters responsible for the resistance of various cancers against chemotherapy. We wanted to study whether MRP2, an ABC transporter of the bile canalicular membrane with a substrate specificity very similar to that of MRP1, is capable of translocating lipids. The translocation of short-chain lipids across the apical membrane of MDCK cells transfected with MRP2 was significantly higher than that in untransfected controls. However, the characteristics of the lipid translocation were similar to substrate transport by MDR1 and not MRP2: transport was strongly inhibited by classic MDR1 Pgp inhibitors, was independent of cellular glutathione, and was insensitive to a drug known to inhibit MRP2 activity. When tested by immunoblot, the MRP2-transfected cells expressed high levels of MRP2 but also of endogenous Mdr1. The expression of Mdr1 was unstable during maintenance of the cell line and correlated with the rate of lipid translocation across the apical membrane. We conclude that the observed increase in lipid transport in the MDCK cells transfected with MRP2 is the consequence of the upregulation of the expression of endogenous Mdr1 and that careful characterization of endogenous Mdr1 expression is needed in studies aimed to identify substrates of plasma membrane transporters. PMID- 11935295 TI - Localization of beta 2-microglobulin in the term villous syncytiotrophoblast. AB - The non-covalent association of beta 2-microglobulin with MHC class I molecules and MHC class I-type molecules such as FcRn or the hemochromatosis protein (HFE) is of major importance for their function, i.e., antigen presentation, IgG transport, and regulation of iron uptake, respectively. In the human hemochorial placenta, the syncytiotrophoblast forms a continuous epithelial layer covering the villous trees, where it directly contacts maternal blood and, among many other functions, mediates uptake of maternal IgG and iron. The villous syncytiotrophoblast lacks MHC class I molecules but expresses FcRn and HFE. Since data on beta 2-microglobulin synthesis and localization in the term villous syncytiotrophoblast were contradictory, we investigated the subcellular localization of beta 2-microglobulin by immunoelectron microscopy. Synthesis in the trophoblast is demonstrated by colocalization of beta 2-microglobulin with protein disulfide isomerase, a marker protein of the endoplasmic reticulum. The presence of beta 2-microglobulin at the apical plasma membrane corresponds to the recently observed association of beta 2-microglobulin with HFE and FcRn. Localization of beta 2-microglobulin in late endosomes/lysosomes, labeled with antibodies to lysosome membrane antigen LAMP 2, suggests also a degradative route of beta 2-microglobulin internalized by fluid-phase from the maternal blood. PMID- 11935297 TI - Comprehensive analysis of promoter methylation and altered expression of hMLH1 in gastric cancer cell lines with microsatellite instability. AB - PURPOSE: Aberrant methylation of the promoter CpG island of hMLH1 is associated with gene silencing in colon cancer and gastric cancer with microsatellite instabilities (MSI). We analyzed the pattern of promoter methylation causing gene silencing. METHODS: Comprehensive analysis of hMLH1 promoter was performed by bisulfite genomic sequencing in human gastric cancer cell lines. Altered expression of hMLH1 was examined by the immunocytochemical staining method and RT PCR, and microsatellite instability was examined using two representative mononucleotide repeat microsatellite markers, BAT-25 and BAT-26. RESULTS: As a result, MSI-positive gastric cancer cell lines were methylated extensively at the overall promoter region. MSI-negative gastric cancer cell lines - except in SNU 620 - were unmethylated completely at the overall promoter region including the more upstream region in contrast to colorectal cancer cell lines. Even though SNU 620 was methylated fully at the overall promoter region - except for partial methylation at the specific region (from -270 to -199) near the transcriptional start - hMLH1 protein was expressed. CONCLUSION: Our data suggest that the methylation density of a specific region plays an important role in gene inactivation of hMLH1 and that the methylation status of the more upstream promoter region and exon 1 start region are not essential for gene inactivation. PMID- 11935298 TI - Molecular characterization of breast cancer cell lines by expression profiling. AB - PURPOSE: Gene expression patterns provide detailed insights into cellular regulation that reflect minor differences of cellular capacity not accessible by standard descriptions of the cellular phenotype or origin. METHODS: To identify fundamental differences and similarities we analyzed the gene expression patterns of four breast cancer cell lines: MCF-7, SK-BR-3, T-47D, and BT-474. RESULTS: Although only a small subset of genes (597) is represented on the Atlas-cDNA Array (Clontech) used, clear differences in the expression of a number of genes could be detected. For example, unique high levels of expressions were found for the HLH-protein ID-1 (MCF-7) and the receptor tyrosine kinase erbB2 (SK-BR-3 and T-47D). Most genes analyzed were expressed at comparable levels in all cell lines studied. CONCLUSIONS: For interpretation of the results sets of genes that show similar variation of expression among the cells were grouped together. Furthermore, our analysis allows the assignment of similarity values that lead to a relation profile of the cell lines. How these results correlate with known biological properties of the cell lines is discussed. Additionally, we demonstrate that results obtained by cDNA-Array hybridization for expression of the ErbB receptor family correlate well with competitive RT-PCR, thus confirming the reliability of the cDNA-Array analysis. PMID- 11935299 TI - Induction of the CBP transcriptional co-activator early during laryngeal carcinogenesis. AB - PURPOSE: CREB-binding protein (CBP) is a transcriptional "integrator" that is suspected of contributing to tumorigenesis. This is the first systematic morphologic study evaluating CBP expression in a large series of human laryngeal tissues containing normal epithelium, premalignant lesions (hyperplasia and/or dysplasia), and squamous cell carcinoma. METHODS: Immunohistochemical methodology was performed on formalin-fixed paraffin-embedded sections by using a monoclonal anti-CBP antibody. CBP expression was screened and compared in 156 patients with various laryngeal histologic entities. RESULTS: Nuclear expression of CBP was found in 44 out of 91 (48.4%) specimens with normal-appearing epithelium (46.2% weak and only 2.2% moderate positivity), 92 out of 100 (92%) with hyperplastic lesions (56% weak, 36% moderate/strong, and only 8% no positivity), 80 out of 103 (77.7%) with dysplastic lesions (45.6% weak, 32.1% moderate/strong, and 22.3% no positivity), 37 out of 45 (82.2%) with well-differentiated carcinoma (42.2% weak, 40% moderate/strong, and 17.8% no positivity), 31 out of 43 (72.1%) with moderately differentiated carcinoma (32.6% weak, 39.5% moderate/strong, and 27.9% no positivity) and eight out of 12 (66.7%) with poorly differentiated carcinoma (41.7% weak, 25% moderate/strong, and 33.3% no positivity). Statistical analysis and correlation of the intensity of nuclear immunostaining among the various histologic entities revealed statistically significant results. CONCLUSIONS: Overexpression of CBP is detected from the very early stages of laryngeal carcinogenesis, suggesting that CBP may play a role in malignant transformation of precancerous laryngeal lesions. It is possible that overexpression of this protein is a prerequisite for the observed p53 upregulation in premalignant lesions, implying an indirect role of CBP in p53-mediated tumorigenic potential. PMID- 11935300 TI - p53 Gene status in relation to ex vivo chemosensitivity of non-small cell lung cancer. AB - PURPOSE: About 40% of non-small cell lung cancer (NSCLC) patients are candidates for systemic chemotherapy, despite the fact that at diagnosis most NSCLC are usually chemoresistant both in vivo and ex vivo. It is important to develop sufficient methods of prediction of the response to chemotherapy and to find molecular markers that may prognose this response. Therefore, a study on the relationship of p53gene status to the ex vivo chemosensitivity of primary human NSCLC was performed. METHODS: Three drug combinations (carboplatin/etoposide, cyclophosphamide/etoposide/epirubicin, and paclitaxel/carboplatin) were tested in a modified ATP cell viability assay. A group of 28 cases of primary human NSCLC was assessed. RESULTS: Ex vivo chemosensitivity testing showed that tumors with p53 mutations were significantly more resistant to the cyclophosphamide/etoposide/epirubicin regimen than with normal p53 gene ( P = 0.012). However, no correlation was observed for two other treatment regimens. CONCLUSION: Mutations in the p53gene can lead to enhanced chemoresistance, confirming the hypothesis that the p53 gene may serve as a marker of tumor response to treatment in NSCLC. However, the data also illustrate that some additional factors might contribute to drug resistance of the examined tumors. PMID- 11935301 TI - Hepatic veno-occlusive disease (VOD) with complete occlusion of liver venules after tandem autologous stem cell transplantation-- successful treatment with high-dose methylprednisolone and defibrotide. AB - Veno-occlusive disease (VOD) is a life-threatening complication following allogeneic or autologous stem cell transplantation. We report on a patient with a high grade B-cell lymphoma who presented 28 days after the second autologous stem cell transplantation with weight gain, ascites, hyperbilirubinemia, and liver venules occlusion as demonstrated by sonography. Starting with high-dose methylprednisolone treatment followed by defibrotide maintenance therapy the patient showed dramatic complete response of VOD, resulting in a normal sonography of the liver and normalization of laboratory values. The response of the occlusion of nearly all liver venules underlines the value of anti inflammatory treatment combined with new thrombolytic medication such as defibrotide for the treatment of severe VOD. PMID- 11935302 TI - The role of neoadjuvant chemotherapy and interval laparotomy in advanced ovarian cancer. AB - Radical debulking surgery allows for optimal cytoreduction in less than 50% of patients with advanced ovarian cancer. In spite of highly efficient chemotherapeutic regimens, the prognosis of patients with residual tumor masses larger than 1 cm in diameter following staging laparotomy is very poor. This observation led to the initiation of numerous trials evaluating the feasibility and efficiency of the use of primary chemotherapy followed by interval laparotomy in women with advanced ovarian cancer. The available data is presented and discussed in this review. PMID- 11935303 TI - Complete remission of a primary cutaneous B-cell lymphoma of the lower leg by first-line monotherapy with the CD20-antibody rituximab. AB - BACKGROUND: Rituximab is a genetically engineered antibody recognizing the CD20 antigen known to be expressed by more than 95% of B-cell lymphomas. Recently the antibody has been approved for routine administration in primary extracutaneous, treatment-refractory or relapsed low-grade, follicular non-Hodgkin B-cell lymphomas. With regard to the pathogenetically related primary cutaneous lymphomas, the so-called large B-cell lymphoma of the leg represents a distinct, but rare subentity. In an 89-year-old, multimorbid patient who was affected by such a non-resectable CD20+ large B-cell lymphoma limited to the skin of both lower legs, rituximab was used as a first-line monotherapy in order to avoid local or systemic toxicities inevitably linked to conventional treatment modalities, i.e., radio- or chemotherapy. METHODS: Rituximab was administered at a dosage of 375 mg/m(2) i.v. eight times in weekly intervals. As a premedication we used prednisolone 150 mg i.v. as well as loratadine 10 mg p.o. 1 h before each rituximab infusion. RESULTS: The treatment was well tolerated without any adverse reactions, but was accompanied by a mild transient blood eosinophilia. The histologically proven, exophytic, multi-nodular lymphoma showed a substantial regression already at 2 weeks after the onset of the rituximab treatment. At 8 weeks we observed a complete clinical remission which is now stabile for a follow up period of 6 months without any maintenance therapy. CONCLUSIONS: Our case observation demonstrates that an intensified, i.e. eightfold, rituximab application in weekly intervals may be a highly effective, tumor target cell specific first-line monotherapy in the management of primary cutaneous large B cell lymphoma of the leg. Given the rareness of the disease, the result as well as the possible contribution of the prednisolone premedication will have to be evaluated in a future, controlled, multi-centre study. PMID- 11935304 TI - Epidermal growth factor receptor expression in invasive thymoma. AB - PURPOSE: Epidermal growth factor receptor (EGFR) is a transmembrane glycoprotein with intrinsic tyrosine kinase activity. Activation results in a variety of cellular responses including cell proliferation and differentiation. In clinical trials, anti-EGFR is showing promise in the treatment of solid tumors expressing EGFR. Thus, we assessed EGFR expression in a series of thymic epithelial tumors. METHODS: Tumors from 37 patients seen at Indiana University School of Medicine (IUMC) for treatment of thymoma (31 patients) or thymic carcinoma (six patients) were assessed for EGFR expression. Five-micron sections of formalin-fixed, paraffin-embedded tumor (28 invasive and/or metastatic thymomas, six thymic carcinomas, and three non-invasive thymomas) were stained with anti-EGFR. Any degree of cytoplasmic membrane staining of tumor cells was considered positive; furthermore, staining was scored 0 to 3+ using criteria as standardized for HER 2/neu assessment of breast carcinoma. Appropriate controls were performed. RESULTS: Positive staining of tumor was observed in 28 tumors (23 invasive and/or metastatic thymomas, two thymic carcinomas, and three non-invasive thymomas). CONCLUSIONS: EGFR is expressed in a high percentage of thymic epithelial tumors. EGFR is often strongly expressed and is a potential therapeutic target in patients with malignant thymic tumors. We are pursuing additional studies to assess anti-EGRF in the treatment of patients with advanced thymoma. PMID- 11935305 TI - An unusual case of malignant gastro-intestinal obstruction in advanced cervical cancer. PMID- 11935306 TI - Aromatase (CYP19) expression in tumor-infiltrating lymphocytes and blood mononuclears. AB - PURPOSE: To clarify the role of lymphocytes as a possible source of estrogens. METHODS: In the present study, lymphocytes were isolated from 11 surgical samples of breast cancer after tumor enzyme digestion and Ficoll/Verographine procedure. Simultaneously, using the latter procedure, mononuclears were separated from the blood of 15 female volunteers. RESULTS: Expression of the aromatase (CYP19) gene was readily demonstrated by standard RT-PCR in blood mononuclears cultivated in the presence of 10% fetal calf serum for 48 h. In the tumor-infiltrating lymphocytes (TIL) of breast cancer patients, CYP19 expression was discovered only with the aid of nested PCR. CONCLUSIONS: The data obtained suggest that aromatase gene expression is presented in TIL at a rather low level. Nevertheless, this can have some functional significance for the estrogen-dependent growth of breast cancer tissue. PMID- 11935307 TI - Immunostimulatory activity of cationic-lipid-nucleic-acid complexes against cancer. AB - Recent studies have highlighted the immunostimulatory nature of nucleic acids and the enhancement of such immunostimulation when nucleic acids are complexed to cationic liposomes to form cationic-lipid-nucleic-acid-complexes or lipoplexes. While such immunostimulation may have deleterious consequences for nucleic acid delivery, especially in the field of gene therapy, it may be harnessed for efficacious usage against the various forms of cancer. PMID- 11935308 TI - Morphologic diagnosis of leukaemic B-lymphoproliferative disorders and the role of cyclin D1 expression. AB - PURPOSE: This study analysed the morphologic differences between leukaemic mantle cell lymphoma, follicular lymphoma, nodal marginal zone lymphoma, and diffuse large B-cell lymphoma in peripheral blood. Additionally, we investigated the role of cyclin D1 expression in B-lymphoproliferative disorders. METHODS: The morphologic analysis of the leukaemic cells was performed on cytocentrifuge preparations after separation of mononuclear cells from peripheral blood using a Ficoll-Hypaque density gradient. Cyclin D1 protein expression was studied with the catalyzed signal amplification system. The expression of other markers (CD5, CD23, light chain immunoglobulins) was analysed by the APAAP method. RESULTS: We describe in detail the morphology of the lymphoma cells in eight patients with mantle cell lymphoma, six patients with follicular lymphoma, 11 patients with nodal marginal zone lymphoma, and seven patients with diffuse large B-cell lymphoma. The morphological distinction between these lymphoma cells is a challenge for the haematologist. The investigation of cytocentrifuge preparations of mononuclear cells allows the detection of lymphoma cells also in cases with nondiagnostic white cell differential. Additionally, the immunotype (light chain restriction, CD5, CD23, and cyclin D1) of 108 patients with leukaemic B lymphoproliferative disorders was studied. Diffuse nuclear expression of cyclin D1 protein (>20%) was specific for mantle cell lymphoma. However, only 6/8 patients showed cyclin-D1 positivity. CONCLUSIONS: The morphologic analysis of lymphoma cells in cytocentrifuge preparations of mononuclear leukocytes in combination with immunocytochemical investigation allows the detection of mantle cells, centrocytes of follicular lymphoma, marginal zone cells, and cells of the diffuse large B-cell lymphoma in peripheral blood. The positivity of cyclin D1 protein improves the differentiation of mantle cells from other lymphoma cells. PMID- 11935309 TI - Transfection of the nm23-H1 gene into human hepatocarcinoma cell line inhibits the expression of sialyl Lewis X, alpha1,3 fucosyltransferase VII, and metastatic potential. AB - PURPOSE: The expressions of Lewis antigens, alpha1,3 fucosyltransferase (alpha1,3 FucT)-VII, and the metastatic potential of the 7721 human hepatocarcinoma cell line after the transfection of the cDNA of nm23-H1, a known metastasis suppressive gene, were studied using mock cells as the control, which were transfected with the pcDNA3 vectors. METHODS: Cell adhesion to human umbilical vein epithelial cells (HUVEC), chemotaxic cell migration through transwells, and invasion through matrigel were selected as the metastasis-related phenotypes to assess the metastatic potential at the cell level. RESULTS: The results showed that the expression of SLe(x) was high, while the expression of Le(x), SDLe(x), and SLe(a) were very low on the surface of the mock cells. After transfection of nm23-H1, the expressions of SLe(x), alpha1,3 FucT-VII, and the cell adhesion to HUVEC, as well as cell migration and invasion were simultaneously decreased in all three clones of nm23-H1-transfected cells. Among different clones, the decreased expressions of SLe(x) and alpha1,3 FucT-VII were roughly correlated to each other, and also, in general, proportional to the ability of cell adhesion to HUVEC, cell migration, and invasion. The expressions of these metastasis-related phenotypes were lowest in clone 3 and highest in clone 4. Only the specific monoclonal antibody to SLe(x) (KM93) significantly abolished the cell adhesion, migration, and invasion, while other monoclonal antibodies against SDLe(x) or Le(x) and SLe(a) only slightly inhibited or entirely failed to inhibit the above mentioned phenotypes. However, the rate of cell growth was not changed after the transfection of nm23-H1, and the ability of colony formation on the soft agar was only decreased in one clone. CONCLUSIONS: These findings reveal that the down regulation of alpha1,3 FucT-VII and its product, SLe(x), is one of the mechanisms to explain the metastasis-suppressive effect of the nm23-H1 gene. PMID- 11935310 TI - Activity of type IV collagenases (MMP-2 and MMP-9) in primary pulmonary carcinomas: a quantitative analysis. AB - PURPOSE: Matrix metalloproteinases MMP-2 and MMP-9 are implicated in invasion and metastasis of malignant tumors. We investigated the expression and activation of MMP-2 and MMP-9 in lung cancer compared with normal lung parenchyma, and looked for a potential marker of malignancy. METHODS: Thirty-six pulmonary carcinomas and paired normal lung specimens were analyzed by gelatin zymography and computer assisted image analysis for the expression of MMP-2 and MMP-9. RESULTS: We showed that expression of both type IV collagenases was remarkably higher in carcinoma samples than in lung parenchyma. The MMP-9 levels in lung cancer were over twofold higher than in normal lung tissues. The levels of latent and active forms of MMP-2 in lung cancer samples were, correspondingly, 3.8- and 17-fold higher than in lung parenchyma. The tumor/normal (T/N) ratios of MMP-2 were negatively correlated with the hemoglobin levels and erythrocytes number. CONCLUSIONS: A high level of the active form of MMP-2 in almost all of the carcinomas and the near lack of its activation in normal lung parenchyma shows that MMP-2 activation is associated with the malignant phenotype and may serve as a good marker of malignancy. The correlation between low hemoglobin level and T/N ratio of MMP-2 may indicate significance of MMP-2 for angiogenesis. PMID- 11935311 TI - Age-dependent differences in tumor cell polarity in endometrial carcinomas. AB - PURPOSE: Cell polarization is dependent on the structural asymmetry of apical and basolateral domains. Although clinical and biological heterogeneity of endometrial carcinomas in aged individuals has been proposed, little is known about the age-dependent differences in tumor morphology in terms of cell polarity. To clarify a possible significance of tumor cell polarity, a total of 92 cases of grade (G) 1 and G2 endometrial carcinomas (endometrioid type) were investigated. METHODS: Cell polarity for tumor glandular components was evaluated by examining the degree of three morphological parameters: nuclear ordering, basal positioning of nuclei within cells, and papillary snouting/glandular convolution. The results were also compared with findings for cell proliferation, expression of p21(Cip1)(p21), p27(Kip1)(p27), estrogen and progesterone receptors, p53 accumulation, and several clinicopathological factors. RESULTS: Average values for each polarity parameter showed a stepwise decrease from tumors of pre-, through peri-, to post-menopausal patients, the difference being significant. Significantly high values for all polarity scores were also evident in tumors adjacent to secretory non-neoplastic endometrium as compared to those in non-secretory tissue. Positive correlations with low cell proliferation determined with respect to Ki-67 labeling indices, early clinical stage, and upper myometrial invasion were also noted, while there were no associations with expression of p21, p27, p53, and two hormone receptors. CONCLUSIONS: These findings indicate that in endometrial carcinomas, the age-dependent differences in tumor cell polarity may be related to status of endogenous ovarian hormones, closely linked with cell proliferation and some prognostic factors. PMID- 11935312 TI - Temozolomide and whole brain irradiation in melanoma metastatic to the brain: a phase II trial of the Cytokine Working Group. AB - PURPOSE: To evaluate the antitumor effects and toxicities of whole brain irradiation (WBI) with temozolomide (TMZ) administered by prolonged oral dosing in patients with melanoma metastatic to the brain. BACKGROUND: Patients with melanoma metastatic to the central nervous system (CNS) have an extremely poor prognosis and appear to benefit little from WBI. TMZ is an alkylating agent chemically similar to dacarbazine (DTIC) with good oral bioavailability and CNS penetration. TMZ has broad preclinical antitumor activity which in melanoma is comparable to that of DTIC. The combination of TMZ and WBI may provide enhanced antitumor activity against CNS metastasis from melanoma. PATIENTS AND METHODS: Patients with measurable CNS metastases with or without systemic disease were treated with WBI, 30 Gray over ten fractions (days 1-5 and 8-12). TMZ, 75 mg small middle dotm(2 small middle dot)day, was started on day 1, continued daily for 6 weeks and repeated every 10 weeks. RESULTS: Thirty-one patients were treated. There was one CNS complete response of 4.5 months and two CNS partial responses of 2 months and 7 months duration; the latter patient also had a 4 month complete remission of systemic metastases. Toxicities were limited to a single episode of grade 3 transaminase elevation and two episodes of grade 3 neutropenia, one complicated by fatal sepsis. The median progression-free interval for both CNS and extracranial sites was 2 months (range 1 week-11 months), and median survival 6 months (range 2-12 months). CONCLUSIONS: WBI has lower than expected activity in CNS metastasis of malignant melanoma. Although TMZ can be safely administered with WBI, the combination has limited anti-tumor activity. PMID- 11935313 TI - Autoantibodies in sera of pancreatic cancer patients identify recombination factor Rad51 as a tumour-associated antigen. AB - PURPOSE: The recombination factor Rad51 is highly expressed in human pancreatic adenocarcinoma. In this study we asked whether high-level expression of Rad51 antigen stimulates a B-cell response leading to Rad51-specific autoantibodies in human pancreatic cancer patients. METHODS: Sera of patients suffering from pancreatic cancer (57) as well as sera of healthy donors (86) were screened for Rad51 autoantibodies by Western-blot analysis. Rad51 over-expressing cell lines were used as antigen source. RESULTS: Four out of 57 (7%) sera tested were found positive for Rad51 autoantibodies of IgG subclass, while all 86 control sera were negative. CONCLUSION: This observation identifies Rad51 as a tumour-associated antigen in pancreatic adenocarcinoma. Since high-level expression of Rad51 is restricted to tumour cells, Rad51 is also a tumour-specific antigen. Further analyses should reveal whether Rad51 might also function as a tumour-specific transplantation antigen (TSTA) and whether it might serve as a target for new immunotherapeutical approaches. PMID- 11935315 TI - Symposium on mathematical, statistical, and computational methods for cancer surveillance and cancer screening. July 27, 2001 Heidelberg, Deutsches Krebsforschungszentrum. PMID- 11935314 TI - Induction of apoptosis by norcantharidin in human colorectal carcinoma cell lines: involvement of the CD95 receptor/ligand. AB - PURPOSE: Cantharidin, a natural toxin, is the active substance of mylabris and has antitumor effects in man. Norcantharidin, the demethylated analogue of cantharidin, has been used in the treatment of patients with primary hepatoma and those with leukopenia in China. The present study was designed to investigate whether norcantharidin exerts cytotoxic activity against colorectal cancer cells by inducing apoptosis and to examine the possible mechanism in the phenomenon. METHODS: Inhibition of proliferation of norcantharidin on Colo205, HT-29, and SW480 colorectal cancer cells was determined by the trypan blue dye exclusion test. Apoptosis of norcantharidin-treated cells was determined by morphological analysis, agarose gel DNA electrophoresis, and quantitated by flow cytometry after staining with propidium iodide. Cell cycle and the cell surface expression of the CD95/CD95 ligand were evaluated by flow cytometry. Caspase 8-like protease and protein phosphatase 1 and 2A activities were also analyzed. RESULTS: Treatment with norcantharidin of colorectal cancer cells not only inhibited cell proliferation, but also induced apoptosis. Norcantharidin induced apoptosis mainly in two phases: rapid apoptosis in S-phase cells and delayed apoptosis in G2/M arrested cells. Treatment with norcantharidin resulted in an upregulation of the CD95 receptor and CD95 ligand on the cell surface. Furthermore, stimulation with anti-CD95 monoclonal antibody (mAb) resulted in further induction of apoptosis after treatment with norcantharidin. In addition, the apoptosis inducing effect of norcantharidin was almost completely inhibited by anti-CD95 ligand mAb. Norcantharidin-treated cells showed the activation of caspase 8. Both zVAD-FMK (a broad range caspase inhibitor) and IETD-FMK (a caspase-8 inhibitor) showed apparent inhibition of the apoptosis-inducing effect. Norcantharidin did not show an inhibitory effect on protein phosphatase. CONCLUSIONS: These results suggest that norcantharidin triggers apoptosis in colorectal cancer cell lines via the activation of the CD95 receptor/ligand system, and that this agent may be useful for developing new therapeutic regimens for the treatment of colorectal carcinoma. PMID- 11935317 TI - Polymorphic GGC repeats in the androgen receptor gene are associated with hereditary and sporadic prostate cancer risk. AB - Androgen receptor (AR) has long been hypothesized to play an important role in prostate cancer etiology. Two trinucleotide repeat polymorphisms (CAG and GGC repeats in exon 1 of the AR gene) have been investigated as risk factors for prostate cancer in several studies. However, the results are inconclusive, probably because of the variations of study designs, characteristics of study samples, and choices of analytical methods. In this study, we evaluated evidence for linkage and association between the two AR repeats and prostate cancer by using the following comprehensive approaches: (1) a combination of linkage and association studies, (2) a test for linkage by parametric analysis and the male limited X-linked transmission/disequilibrium test (XLRC-TDT), (3) a test for association by using both population-based and family-based tests, and (4) a study of both hereditary and sporadic cases. A positive but weak linkage score (HLOD=0.49, P=0.12) was identified in the AR region by parametric analysis; however, stronger evidence for linkage in the region, especially at the GGC locus, was observed in the subset of families whose proband had < or = 16 GGC repeats (HLOD=0.70, P=0.07) or by using XLRC-TDT ( z'=2.65, P=0.008). Significantly increased frequencies of the < or = 16 GGC repeat alleles in 159 independent hereditary cases (71%) and 245 sporadic cases (68%) cases compared with 211 controls (59%) suggested that GGC repeats were associated with prostate cancer ( P=0.02). Evidence for the association between the < or = 16 GGC repeats and prostate cancer risk was stronger with XLRC-TDT ( z'=2.66, P=0.007). No evidence for association between the CAG repeats and prostate cancer risk was observed. The consistent results from both linkage and association studies strongly implicate the GGC repeats in the AR as a prostate cancer susceptibility gene. Further studies on this polymorphism in other independent data sets and functional analysis of the GGC repeat length on AR activity are warranted. PMID- 11935316 TI - A genomic map of a 6-Mb region at 13q21-q22 implicated in cancer development: identification and characterization of candidate genes. AB - Chromosomal region 13q21-q22 harbors a putative breast cancer susceptibility gene and has been implicated as a common site for somatic deletions in a variety of malignant tumors. We have built a complete physical clone contig for a region between D13S1308 and AFM220YE9 based on 18 yeast artificial chromosome and 81 bacterial artificial chromosome (BAC) clones linked together by 22 genetic markers and 61 other sequence tagged sites. Combining data from 47 sequenced BACs (as of June 2001), we have assembled in silico an integrated 5.7-Mb genomic map with 90% sequence coverage. This area contains eight known genes, two hypothetical proteins, 24 additional Unigene clusters, and approximately 100 predicted genes and exons. We have determined the cDNA and genomic sequence, and tissue expression profiles for the KIAA1008 protein (homologous to the yeast mitotic control protein dis3+), KLF12 (AP-2 repressor), progesterone induced blocking factor 1, zinc finger transcription factor KLF5, and LIM domain only-7, and for the hypothetical proteins FLJ22624 and FLJ21869. Mutation screening of the five known genes in 19 breast cancer families has revealed numerous polymorphisms, but no deleterious mutations. These data provide a basis and resources for further analyses of this chromosomal region in the development of cancer. PMID- 11935318 TI - The role of mtDNA background in disease expression: a new primary LHON mutation associated with Western Eurasian haplogroup J. AB - Leber's hereditary optic neuropathy (LHON) is a maternally transmitted form of blindness caused by mitochondrial DNA (mtDNA) mutations. Approximately 90% of LHON cases are caused by 3460A, 11778A, or 14484C mtDNA mutations. These are designated "primary" mutations because they impart a high risk for LHON expression. Although the 11778A and 14484C mutations unequivocally predispose carriers to LHON, they are preferentially associated with mtDNA haplogroup J, one of nine Western Eurasian mtDNA lineages, suggesting a synergistic and deleterious interaction between these LHON mutations and haplogroup J polymorphism(s). We report here the characterization of a new primary LHON mutation in the mtDNA ND4L gene at nucleotide pair 10663. The homoplasmic 10663C mutation has been found in three independent LHON patients who lack a known primary mutation and all of which belong to haplogroup J. This mutation has not been found in a large number of haplotype-matched or non-haplogroup-J control mtDNAs. Phylogenetic analysis with primarily complete mtDNA sequence data demonstrates that the 10663C mutation has arisen at least three independent times in haplogroup J, indicating that it is not a rare lineage-specific polymorphism. Analysis of complex I function in patient lymphoblasts and transmitochondrial cybrids has revealed a partial complex I defect similar in magnitude to the 14484C mutation. Thus, the 10663C mutation appears to be a new primary LHON mutation that is pathogenic when co occurring with haplogroup J. These results strongly support a role for haplogroup J in the expression of certain LHON mutations. PMID- 11935319 TI - Relaxation of imprinted expression of ZAC and HYMAI in a patient with transient neonatal diabetes mellitus. AB - Transient neonatal diabetes mellitus (TNDM) is a rare disease believed to result from overexpression of a paternally expressed gene controlled by a differentially methylated CpG island on chromosome 6q24. Two genes partially overlap the island: the cell-cycle-control gene ZAC and the untranslated gene HYMAI, the function of which is currently unknown. Proof that either gene is involved in TNDM would require demonstration that imprinted expression is relaxed in TNDM patients; this has hitherto been lacking because of the rarity of the disease and the lack of imprinted expression in the lymphoblastoid cells that are generally the only resource available for study. Here, we show, for the first time, the aberrant expression of imprinted genes in a TNDM patient. In TNDM fibroblasts, the monoallelic expression of both ZAC and HYMAI is relaxed, providing strong supportive evidence that the presence of two unmethylated alleles of this locus is indeed associated with the inappropriate gene expression of neighbouring genes. PMID- 11935320 TI - Linkage analysis identifies the thyroglobulin gene region as a major locus for familial congenital hypothyroidism. AB - Congenital hypothyroidism affects 1/3000-4000 newborns and it has been estimated that 10-20% are familial cases with an autosomal recessive mode of inheritance. Previous studies of mostly individual cases have led to the identification of mutations in a number of genes, indicating that it is a genetically heterogeneous disease, but no major gene has been identified. In the present investigation, a population-based sample of 23 families with autosomal recessive congenital hypothyroidism, but no signs of goitre, were subject to linkage analysis. When markers located close to the thyroglobulin gene on chromosome 8q24 were used in a two-point analysis allowing for heterogeneity, a Z(max) of 4.10 was obtained with the microsatellite marker D8S557, indicating heterogeneity with 43% of the families being linked. A multipoint analysis using the markers D8S557 and D8S1835 gave a Z(max) of 3.51, assuming homogeneity. There was significant evidence of heterogeneity with 44.5% of the families being linked. The results indicate that a gene in 8q24 is a common cause of familial congenital hypothyroidism. Since thyroglobulin is essential for thyroid physiology, the gene encoding this protein is the obvious candidate for mutation analysis in the linked families. PMID- 11935321 TI - Effectiveness of computational methods in haplotype prediction. AB - Haplotype analysis has been used for narrowing down the location of disease susceptibility genes and for investigating many population processes. Computational algorithms have been developed to estimate haplotype frequencies and to predict haplotype phases from genotype data for unrelated individuals. However, the accuracy of such computational methods needs to be evaluated before their applications can be advocated. We have experimentally determined the haplotypes at two loci, the N-acetyltransferase 2 gene ( NAT2, 850 bp, n=81) and a 140-kb region on chromosome X ( n=77), each consisting of five single nucleotide polymorphisms (SNPs). We empirically evaluated and compared the accuracy of the subtraction method, the expectation-maximization (EM) method, and the PHASE method in haplotype frequency estimation and in haplotype phase prediction. Where there was near complete linkage disequilibrium (LD) between SNPs (the NAT2 gene), all three methods provided effective and accurate estimates for haplotype frequencies and individual haplotype phases. For a genomic region in which marked LD was not maintained (the chromosome X locus), the computational methods were adequate in estimating overall haplotype frequencies. However, none of the methods was accurate in predicting individual haplotype phases. The EM and the PHASE methods provided better estimates for overall haplotype frequencies than the subtraction method for both genomic regions. PMID- 11935323 TI - Evidence of a founder effect and refinement of the hereditary neuralgic amyotrophy (HNA) locus on 17q25 in American families. AB - Hereditary neuralgic amyotrophy (HNA) is an autosomal dominant disorder that is associated with episodic recurrent brachial plexus neuropathy. A mutation for HNA maps to chromosome 17q25. To refine the HNA locus further, we carried out genetic linkage studies in seven pedigrees with a high density set of DNA markers from chromosome 17q25. All pedigrees demonstrated linkage to chromosome 17q25, and an analysis of recombinant events placed the HNA locus within an interval of approximately 1 Mb flanked by markers D17S722 and D17S802. In order to test the power of linkage disequilibrium mapping, we compared genotypes of 12 markers from seven pedigrees that were from the United States and that showed linkage to chromosome 17q25. The haplotypes identified a founder effect in six of the seven pedigrees with a minimal shared haplotype that further refines the HNA locus to an interval of approximately 500 kb. These findings suggest that, for the pedigrees from the United States, there are at least two different mutations in the HNA gene. PMID- 11935322 TI - Identification, genomic organization, chromosomal mapping and mutation analysis of the human INV gene, the ortholog of a murine gene implicated in left-right axis development and biliary atresia. AB - Determination of left-right axis is a precocious embryonic event, and all phenotypic anomalies resulting from disruption of the normal lateralization process are collectively referred to as the lateralization defect. A transgenic mouse with lateralization defect and hepatic, kidney, and pancreatic anomalies has resulted from disruption of the inv gene by insertion of a transgene. The human ortholog is thus a good candidate for lateralization defect in humans, in particular in cases with associated hepatic anomalies. Here, we have identified, mapped, and characterized the INV human gene and screened a series of heterotaxic patients (with or without biliary anomalies) for mutation in this gene. In a German family of Turkish origin, we have found that all available affected and unaffected individuals are heterozygous for a mutation in the splicing donor site of intron 12 in the INV gene resulting in two different aberrant splicing isoforms. This can be explained either by a randomization of lateralization defects or, as suggested earlier, di- or trigenic inheritance, although we have been unable to detect, in this family, a mutation in genes known to be involved in the human lateralization defect ( LEFTY1, LEFTY2, ACVR2B, NODAL, ZIC3, and CFC1). In contrast to the mouse, the affected individuals have no biliary anomalies, and the absence of mutation in a series of seven cases with lateralization defect and biliary anomalies demonstrates that INV is not frequently involved in such a phenotype in humans. PMID- 11935324 TI - Evidence of multiple causal sites affecting weight in the IGF2-INS-TH region of human chromosome 11. AB - The subtelomeric region of 11p harbours three closely linked genes, TH, INS and IGF2, that have been associated with obesity, size at birth, type I diabetes, polycystic ovary syndrome, overgrowth in Beckwith-Wiedemann syndrome and possibly hypertension. We have previously shown that the IGF2 ApaI single nucleotide polymorphism (SNP) associates with weight and body mass index in middle-aged Caucasian males but that there is no such association with the INS -23/ HphI site that marks INS 5' variable number of tandem repeats (VNTR) class I vs class III VNTR alleles. We report here the examination of three SNP markers in IGF2: 6815 A/T in the P1 promoter, AluI in exon 3 and ApaI in the 3' untranslated region (UTR), INS 5'VNTR class I alleles and the TH01 tetranucleotide microsatellite in a population sample. The analysis has taken into account the possibility that typing failure and the number of parameters required to model multiallelic loci could create spurious significance. We have exercised Hardy-Weinberg equilibrium tests, dichotomised multiallelic series to impose parsimony, and examined the data with failures modelled or excluded. Regression analysis infers that three markers, IGF2 ApaI, TH01 and subclasses of INS VNTR class I independently predict derived weight indices (combined P<10(-8) and accounting up to 2% of population weight variance), with no evidence of interaction. This establishes that there must be multiple causal sites impacting on weight in this genomic region. PMID- 11935325 TI - Epstein syndrome: another renal disorder with mutations in the nonmuscle myosin heavy chain 9 gene. AB - Epstein syndrome (EPTS) is an autosomal dominant disease characterized by nephritis, mild hearing loss, and thrombocytopenia with giant platelets. Renal and hearing abnormalities are indistinguishable from those observed in Fechtner syndrome (FTNS), an Alport-like variant. EPTS macrothrombocytopenia is similar to that described in FTNS, May-Hegglin anomaly (MHA), and Sebastian syndrome (SBS), three disorders caused by mutations in the nonmuscle heavy chain myosin IIA ( MYH9). Unlike FTNS, MHA, and SBS, EPTS does not show inclusion bodies in the leukocytes. The clinical features of EPTS and the chromosomal localization of the respective gene in the same region as MYH9 suggest that this disorder is allelic with the other giant platelet disorders. We identified a MYH9 missense mutation in two EPTS familial cases. In both families, an R702H substitution was found, probably inducing conformational changes to the myosin head. A different amino acid substitution at the same codon (R702C) has been previously identified in FTNS. On the basis of predictions from molecular modeling of the X-ray crystallographic structure of chick smooth muscle myosin, the mutated thiol reactive group of R702C may lead to intermolecular disulfide bridges, with the consequent formation of the inclusions typical of FTNS. On the contrary, the R702H mutation does not allow the protein to aggregate and thus to generate "Dohle-like" bodies, which are indeed absent in EPTS. In conclusion, our results extend the allelic heterogeneity of MYH9 mutations to another clinical syndrome and contribute to the clarification of the pathogenesis of the various inherited giant platelet disorders. PMID- 11935326 TI - Pyruvate dehydrogenase E3 binding protein deficiency. AB - Primary defects of the E3 binding protein component of the pyruvate dehydrogenase complex appear to be a rare cause of pyruvate dehydrogenase deficiency. We describe two new, unrelated patients with mutations in the E3 binding protein gene, in both cases involving the conserved dinucleotides of splice junctions. Both patients presented with delayed development and lactic acidosis, features that are also found in patients with the more common pyruvate dehydrogenase E1 alpha subunit deficiency; however, they both had significant residual enzyme activity in cultured fibroblasts and prolonged survival. PMID- 11935327 TI - Susceptibility gene for non-obstructive azoospermia located near HLA-DR and -DQ loci in the HLA class II region. AB - The technical developments and expanded indications for testicular sperm extraction (TESE) with intracytoplasmic sperm injection (ICSI) provide great advantages for patients with non-obstructive azoospermia. Such success, however, also means that genetic abnormalities in non-obstructive azoospermia can be transmitted to the next generation, demonstrating the importance of being able to understand the genetic background of non-obstructive azoospermia. We have previously reported that human leukocyte antigens (HLA)-A33 and -B44 in the HLA class I region and the HLA-DRB1*1302 allele in the HLA class II region are linked to susceptibility to non-obstructive azoospermia in Japanese men. However, strong linkage of HLA-DRB1*1302 with HLA-A33 and -B44 is also evident in the Japanese population. Thus, uncertainty prevails as to whether the HLA class I or class II molecule is more directly associated with non-obstructive azoospermia. In the present study, we performed association analysis with 21 polymorphic microsatellite markers identified near the HLA genes to map the gene involved in the development of non-obstructive azoospermia more precisely. Microsatellite markers located in the HLA class I region or the class III region showed no statistically significant association with this disorder, although once again the HLA-A33 and -B44 alleles showed a significant association. In contrast, some of the microsatellite markers in the HLA class II region and at the HLA-DRB1 and DQB1 loci displayed strong associations with non-obstructive azoospermia. Taken together, our previous and present data suggest that the critical region for development of non-obstructive azoospermia is near the HLA-DRB1 and -DQB1 segments in the HLA class II region. PMID- 11935328 TI - Y-chromosomal evidence for a strong reduction in male population size of Yakuts. PMID- 11935329 TI - ZNT4 gene is not responsible for acrodermatitis enteropathica in Japanese families. PMID- 11935332 TI - Proposed guidelines for papers describing DNA polymorphism-disease associations. AB - Studies reporting DNA polymorphism-disease associations represent an important source of information on disease gene candidacy. Such studies are, however, extremely variable in terms of their design and of the statistical methodology employed. Guidelines are therefore proposed that are intended to promote the publication of scientifically meaningful disease association studies through the introduction of sensible methodological standards. PMID- 11935333 TI - Origin and affinities of indigenous Siberian populations as revealed by HLA class II gene frequencies. AB - Gene frequencies of eight Siberian populations (Mansi, Tuva, Todja, Tofalar, Buryat, Okhotsk Evenki, Ulchi, and Negidal) were determined for the three most polymorphic HLA class II loci ( DRB1, DQA1, and DQB1) by a combination of single stranded conformational polymorphism typing and DNA sequencing. The number of alleles per population ranged from 16 to 25, from seven to eight, and from nine to 14 for the DRB1, DQA1, and DQB1 loci, respectively. The alleles at the three loci occurred in 66 different combinations (haplotypes), most of which appeared to be of ancient origin, but some may have arisen within the Siberian populations. Phylogenetic analysis of the frequency data suggests that the HLA genes of Asian and indigenous American populations stem from a single pool distinct from the gene pools of European and African populations. The Asian populations separate into two clusters, one of which encompasses nearly all the Siberian populations and all the indigenous American populations tested, while the other consists of Central, Eastern, and Southeastern Asian populations. The position of the Tuva people appears to be near the node from which the two clusters diverge. The divergence time of the two clusters is estimated to be 21,000-24,000 years BP. Three different branches of the native Siberian peoples seem to have contributed founders for the indigenous American ethnic groups. PMID- 11935334 TI - Characterisation of interstitial duplications and triplications of chromosome 15q11-q13. AB - Chromosome 15 is frequently involved in the formation of structural rearrangements. We report the molecular characterisation of 16 independent interstitial duplications, including those of one individual who carried a duplication on both of her chromosomes 15, and three interstitial triplications of the Prader-Willi/Angelman syndrome critical region (PWACR). In all probands except one, the rearrangement was maternal in origin. In one family, the duplication was paternal in origin, yet appeared to segregate in a sibship of three with an abnormal phenotype that included developmental delay and a behavioural disorder. Ten duplications were familial, five de novo and one unknown. All 16 duplications, including two not visible by routine G-banding, were of an almost uniform size and shared the common deletion breakpoints of Prader-Willi syndrome and Angelman syndrome. Like deletions, the formation of duplications can occur in both male and female meiosis and involve both inter- and intrachromosomal events. This implies that at least some deletions and duplications are the reciprocal products of each other. We observed no instances of meiotic instability in the transmission of a duplication, although recombination within the PWACR occurred in two members of the same family between the normal and the duplicated chromosome 15 homologues. All three triplications arose de novo and included alleles from both maternal chromosomes 15. Triplication breakpoints were more variable and extended distally beyond the PWACR. The molecular characteristics of duplications and triplications suggest that they are formed by different mechanisms. PMID- 11935335 TI - Identification and characterization of a novel liver-specific enhancer of the human phenylalanine hydroxylase gene. AB - The liver-specific phenylalanine hydroxylase catalyzes the conversion of phenylalanine to tyrosine. Genetic defects in the gene result in the autosomal recessive disorder phenylketonuria. We have identified a phenylalanine hydroxylase mutation, designated as -4173_-407del, in a hyperphenylalaninemic patient with a 3.7-kb deletion in the 5'-flanking region of the phenylalanine hydroxylase gene. Characterization of the deleted sequence has led to the identification of a novel liver-specific DNase I hypersensitive site located 3.3 kb upstream of the RNA initiation site of the phenylalanine hydroxylase gene. We show that this site comprises a liver-specific enhancer with cAMP responsiveness. We further show by mutational analysis that the enhancer carries a major hepatocyte nuclear factor-1-binding site important for the enhancer function but not for cAMP responsiveness. In transient transfection assays with a reporter gene, we demonstrate that a phenylalanine hydroxylase plasmid construct carrying the -4173_-407del mutation is severely impaired in phenylalanine hydroxylase transcriptional activity. Our data indicate that the 3.7-kb deletion uncovered in the genomic DNA of the phenylketonuria proband is linked to the observed phenylketonuria phenotype as a result of a deletion of the newly identified liver specific enhancer. Our systematic approach to the analysis and subsequent discovery of the novel deletion mutation may be applicable to the search for other mutations in the 5' regulatory region of phenylalanine hydroxylase and other genes. PMID- 11935336 TI - A candidate gene for congenital bilateral isolated ptosis identified by molecular analysis of a de novo balanced translocation. AB - Ptosis is defined as drooping of the upper eyelid and can impair full visual acuity. It occurs in a number of forms including congenital bilateral isolated ptosis, which may be familial and for which two linkage groups are known on chromosomes 1p32-34.1 and Xq24-27.1. We describe the analysis of the chromosome breakpoints in a patient with congenital bilateral isolated ptosis and a de novo balanced translocation 46,XY,t(1;8)(p34.3;q21.12). Both breakpoints were localized by fluorescence in situ hybridisation with yeast artificial chromosomes, bacterial artificial chromosomes and P1 artificial chromosomes. The derived chromosomes were isolated by flow-sorting, amplified by degenerate oligonucleotide-primed polymerase chain reaction and analyzed by sequence tagged sites amplification to map the breakpoints at a resolution that enabled molecular characterization by DNA sequencing. The 1p breakpoint lies ~13 Mb distal to the previously reported linkage locus at 1p32-1p34.1 and does not disrupt a coding sequence, whereas the chromosome 8 breakpoint disrupts a gene homologous to the mouse zfh-4gene. Murine zfh-4 codes for a zinc finger homeodomain protein and is a transcription factor expressed in both muscle and nerve tissue. Human ZFH-4 is therefore a candidate gene for congenital bilateral isolated ptosis. PMID- 11935337 TI - A case of segmental paternal isodisomy of chromosome 14. AB - Uniparental disomy of chromosome 14 (UPD 14) results in one of two distinct abnormal phenotypes, depending upon the parent of origin. This discordance may result from the reciprocal over-expression and/or under-expression of one or more imprinted genes. We report a case of segmental paternal isodisomy for chromosome 14 with features similar to those reported in other paternal disomy 14 cases. Microsatellite marker analysis revealed an apparent somatic recombination event in 14q12 leading to proximal biparental inheritance, but segmental paternal uniparental isodisomy distal to this site. Analysis of monochromosomal somatic cell hybrids containing either the paternally inherited or the maternally inherited chromosome 14 revealed no deletion of the maternally inherited chromosome 14 and demonstrated the presence of paternal sequences from D14S121 to the telomere on both chromosomes 14. Thus, the patient has paternal isodisomy for 14q12-14qter. Because the patient shows most of the features associated with paternal disomy 14, this supports the presence of the imprinted domain(s) distal to 14q12 and suggests that the proximal region of chromosome 14 does not contain imprinted genes that contribute significantly to the paternal UPD 14 phenotype. PMID- 11935338 TI - Genetic study of SMA patients without homozygous SMN1 deletions: identification of compound heterozygotes and characterisation of novel intragenic SMN1 mutations. AB - Autosomal recessive spinal muscular atrophy (SMA) is a disease resulting from mutations in the telomeric survival motor neuron gene ( SMN1). In our sample of 150 Spanish SMA families, 87% of patients had homozygous deletions of SMN1. To identify patients who retained a single SMN1 copy, SMN dosage analysis was performed by a fluorescent quantitative PCR assay. In five out of 19 patients tested we detected one SMN1 copy. An extensive SMN gene analysis in these patients led to identification of four intragenic mutations, including two novel ones: a frameshift mutation in exon 6 (773insC) and a splice site mutation in intron 6 (c.867+2T-->G). Two previously described mutations were also found: a deletion in exon 3 (430del4), identified in several Spanish patients, and a frequently occurring mutation in exon 6 (813ins/dup11), reported in several populations. Although the spectrum of intragenic mutations is small, only 27 reported up to now, identification of three mutations found exclusively in the Spanish population indicates that the occurrence of different intragenic mutations depends on the ethnic origin of SMA patients. In the remaining patient, who had a single SMN1 copy and three SMN2 copies, we found that the SMN1 allele was non-functional; the patient did not show any SMN1 transcript. Sequencing of the SMN promoter regions revealed various differences between promoters of the patient's four SMN genes, in particular a change in the length of a polyA run removing a putative YY1 binding site, which may affect the expression of SMN genes. PMID- 11935339 TI - CIA30 complex I assembly factor: a candidate for human complex I deficiency? AB - The human mitochondrial NADH:ubiquinone oxidoreductase (complex I), the first complex of the oxidative phosphorylation system, is composed of at least 42 subunits. Little is known about the assembly process of these subunits into the mature complex. Recently, two proteins in Neurospora crassa have been found to be involved in the assembly of complex I. These proteins are not constituent parts of the mature complex but are associated with smaller intermediate complexes of the assembly process and have a chaperone-like function. We have characterized the human homologue of one of these two complex I intermediate associated proteins, named CIA30, and show that expression of the human CIA30 protein is ubiquitous with a slightly higher expression in various heart tissues, kidney, lung and liver. As deletion of the Neurospora crassa CIA genes results in severe disruption of the assembly process, human CIA30 can be considered as a candidate gene related to complex I deficiency. Thirteen patients with an isolated complex I deficiency, but who were ruled out for mutations in the 35 nuclear genes of the complex and mtDNA, were subjected to mutational analysis of the gene coding for the human CIA30 protein. Four new single nucleotide polymorphisms (SNPs) were detected but no functional mutation was found. PMID- 11935340 TI - Variability of X chromosome inactivation: effect on levels of TIMP1 RNA and role of DNA methylation. AB - X chromosome inactivation results in dosage equivalency for X-linked gene expression between males and females. However, some X-linked genes show variable X inactivation, being expressed from the inactive X in some females but subject to inactivation in other women. The human tissue inhibitor of metalloproteinases 1 ( TIMP1) gene falls into this category. As TIMP1 and its target metalloproteinases are involved in many biological processes, women with elevated TIMP1 expression may exhibit different disease susceptibilities. To address the potential impact of variable X inactivation, we analyzed TIMP1 expression levels by using an RNase protection assay. The substantial variation of TIMP1 expression observed in cells with monoallelic TIMP1 expression precluded analysis of the contribution of the inactive X to total TIMP1 RNA levels in females, so we examined expression in rodent/human somatic cell hybrids. TIMP1 expression levels varied more widely in hybrids retaining an inactive X than in those with an active X chromosome, suggesting variable retention of the epigenetic silencing mechanisms associated with X inactivation. Therefore, we investigated the contribution of methylation at the promoter to expression level variation and found that methylation of the TIMP1 promoter correlated with instability and low level expression, whereas stable TIMP1expression from the inactive X equivalent to that seen from the active X chromosome was observed when the promoter was unmethylated. Since all female cell lines examined showed methylation of the TIMP1 promoter, the contribution of expression from the inactive X appears minimal. However, as women age, they may accumulate cells stably expressing TIMP1 from the inactive X, with a resulting increase of TIMP1, which may explain some sex differences in various late-onset disorders. PMID- 11935341 TI - Identification of novel mutations in MLC1 responsible for megalencephalic leukoencephalopathy with subcortical cysts. AB - Megalencephalic leukoencephalopathy with subcortical cysts (MLC) is an inherited neurologic disorder with macrocephaly before the age of one and slowly progressive deterioration of motor functions. Magnetic resonance imaging shows diffusely abnormal and swollen white matter of the cerebral hemispheres and the presence of subcortical cysts in the anterior-temporal region and often also in the frontoparietal region. Mutations in the MLC1 gene, encoding a putative membrane protein, have been recently identified as a cause for MLC. Here, we describe 14 new mutations in 18 patients. Two identified polymorphisms lead to alterations of amino acid residues. The role, suggested by others, of a mutation in the MLC1gene in catatonic schizophrenia and the possible function of the MLC1 protein as a cation channel are discussed. PMID- 11935342 TI - Genetics of congenital deafness in the Palestinian population: multiple connexin 26 alleles with shared origins in the Middle East. AB - In some Palestinian communities, the prevalence of inherited prelingual deafness is among the highest in the world. As an initial step towards understanding the genetic causes of hearing loss in the Palestinian population, 48 independently ascertained probands with non-syndromic hearing loss were evaluated for mutations in the connexin 26 gene. Of the 48 deaf probands, 11 (23%) were homozygous or compound heterozygous for mutations in GJB2. Five different mutations were identified: ivs1(+1) G-->A, 35delG, 167delT, T229C, 235delC. Nine deaf probands were homozygous and only two compound heterozygous. Among 400 hearing Palestinian controls, one carrier was observed (for 167delT). We show that GJB2 ivs1(+1) G- >A disrupts splicing, yielding no detectable message. Linkage disequilibrium analysis suggests, in the Palestinian and Israeli populations, a common origin of the 35delG mutation, which is worldwide, and of 167delT, which appears specific to Israeli Ashkenazi and Palestinian populations. A high prevalence of deafness, high frequency of homozygosity rather than compound heterozygosity among deaf, and low mutation carrier frequency together reflect the high levels of consanguinity of many extended Palestinian families. Some of the 25 families with multiple cases of inherited prelingual deafness and wildtype GJB2 sequences may represent as-yet-unknown genes for inherited hearing loss. PMID- 11935347 TI - The systems V(IV)O(2+)-glutathione and related ligands: a potentiometric and spectroscopic study. AB - The equilibria in the system V(IV)O(2+)-glutathione in aqueous solution were studied in the pH range 2-11 by a combination of pH-potentiometry and spectroscopy (EPR, visible absorption and circular dichroism). The results of the various methods are consistent and the equilibrium model includes the species MLH(3), MLH(2), MLH, ML(2)H(2), MLH(-1), and MLH(-2) and several hydrolysis products (where H(4)L denotes totally protonated glutathione); individual formation constants and spectra are given. ML(2)H(2) is the predominant species at physiological pH. Plausible structures for each stoichiometry are discussed. The related V(IV)O(2+) systems of S-methylglutathione and gamma- L-glutamyl- L cysteinyl ethyl ester were studied by means of the same spectroscopic techniques in order to support the established binding modes for the glutathione complexes. The importance of glutathione and oxidized glutathione in binding V(IV)O(2+) in cells is assessed. PMID- 11935348 TI - Stability and transmetallation of the magnetic resonance contrast agent MnDPDP measured by EPR. AB - MnDPDP [manganese(II) N, N'-dipyridoxylethylenediamine- N, N'-diacetate-5,5' bis(phosphate)] is the active component of Teslascan, a contrast medium for magnetic resonance imaging of the liver. It has previously been shown that MnDPDP is rapidly dephosphorylated to the monophosphate MnDPMP and the non phosphorylated MnPLED, and that all these substances are rapidly transmetallated to the corresponding Zn complexes. In the present study we used EPR at 9 and 230 GHz to show that no free Mn(2+) ions can be detected in the product or in a mixture of MnDPDP and human serum. Competition experiments between MnDPDP and Zn(2+), Ca(2+), and Mg(2+) ions revealed approximately 15% transmetallation with Zn(2+) in a buffer system containing metal ion concentrations similar to that in serum, whereas approximately 10% transmetallation was obtained with Ca(2+) and only negligible transmetallation was obtained with Mg(2+) under these conditions. Binding experiments with Mn(2+) added to human albumin and human serum indicate that albumin accounts for most of the protein-bound Mn(2+) in serum. PMID- 11935349 TI - Copper bis(thiosemicarbazone) complexes as hypoxia imaging agents: structure activity relationships. AB - Copper(II) bis(thiosemicarbazone) complexes labelled with Cu-60/62/64 are useful radiopharmaceuticals for imaging blood flow and hypoxic tissues in vivo. The aim of this study was to identify structure-activity relationships within a series of analogues with different alkyl substitution patterns in the ligand, in order to design improved hypoxia imaging agents and elucidate hypoxia selectivity mechanisms. Thirteen such complexes were synthesised and characterised spectroscopically and electrochemically. The uptake of each (labelled with Cu-64) in EMT6 tumour cells in vitro under normoxic and hypoxic conditions was studied. All complexes were taken up efficiently into cells, and some showed strong hypoxia selectivity, which was highly correlated with the Cu(II/I) redox potential. Redox potentials at the low end of the range were found to be essential for hypoxia selectivity. In turn, the redox potential was strongly correlated with alkyl substitution pattern, and the most important determinant of the redox potential was the number of alkyl groups on the diimine backbone of the ligand. Several complexes in the series warrant further evaluation as hypoxia imaging agents. The radioactivity uptake/release behaviour in the cells provides insight into possible mechanisms, and a model for hypoxia-selective intracellular trapping is discussed. PMID- 11935350 TI - Characterization of Hydrogenobacter thermophilus cytochromes c(552 )expressed in the cytoplasm and periplasm of Escherichia coli. AB - Hydrogenobacter thermophilus cytochrome c(552) ( Ht cyt c(552)) is a small monoheme protein in the cytochrome c(551) family. Ht cyt c(552) is unique because it is hypothesized to undergo spontaneous cytoplasmic maturation (covalent heme attachment) when expressed in Escherichia coli. This is in contrast to the usual maturation route for bacterial cytochromes c that occurs in the cellular periplasm, where maturation factors direct heme attachment. Here, the expression of Ht cyts c(552) in the periplasm as well as the cytoplasm of E. coli is reported. The products are characterized by absorption, circular dichroism, and NMR spectroscopy as well as mass spectrometry, proteolysis, and denaturation studies. The periplasmic product's properties are found to be indistinguishable from those reported for protein isolated from Ht cells, while the major cytoplasmic product exhibits structural anomalies in the region of the N-terminal helix. These anomalies are shown to result from the retention of the N-terminal methionine in the cytoplasmic product, and not from heme attachment errors. The (1)H NMR chemical shifts of the heme methyls of the oxidized ( S=1/2) expression products display a unique pattern not previously reported for a cytochrome c with histidine-methionine axial ligation, although they are consistent with native like heme ligation. These results support the hypothesis that proper heme attachment can occur spontaneously in the E. coli cytoplasm for Ht cyt c(552). PMID- 11935352 TI - A theoretical study of the mechanism for peptide hydrolysis by thermolysin. AB - The catalytic mechanism for peptide hydrolysis by thermolysin has been investigated using the B3LYP hybrid density functional method. The starting structure for the calculations was based on the X-ray crystal structure of the enzyme inhibited with the ZF (p)LA phosphonamidate transition-state analogue. Besides the three Zn ligands His142, His146 and Glu166, a few additional residues were also included in the model. Following the order of importance, the outer sphere ligands Glu143, His231 and Asp226 were shown to play significant catalytic roles, well correlated with results from site-directed mutagenesis experiments. A single-step reaction mechanism was obtained starting from the initial enzyme substrate complex with a pentacoordinated metal center and proceeding to the enzyme-carboxylate complex as a final product, following a proposal by Matthews and co-workers. The transition state combines a nucleophilic water oxygen attack on the peptide carbon and a proton transfer from the water to the peptide nitrogen, mediated by the Glu143 carboxylate. A free activation energy of 15.2 kcal/mol was obtained, compared to the experimental 12.4-16.3 kcal/mol range for various peptide substrates. An interesting aspect of the present single-step mechanism is that the Glu143 carboxylate moves a significant distance of ~1.0 A. Different chemical models were examined, both related to the system size and proper side-chain modeling. The significance of the protein frame rigidity around the active site was estimated by fixing and subsequently releasing the edge atom positions. Finally, alternative mechanistic proposals are briefly summarized. PMID- 11935351 TI - Spectroscopic studies on human serum albumin and methemalbumin: optical, steady state, and picosecond time-resolved fluorescence studies, and kinetics of substrate oxidation by methemalbumin. AB - The nature of the heme environment in methemalbumin, the Fe(III) protoporphyrin IX (heme)-human serum albumin (HSA) complex, was investigated by optical spectroscopy. Comparison of the optical spectra of methemalbumin, ferro hemalbumin in the absence and presence of 2-methylimidazole, and their carbon monoxide derivatives with horseradish peroxidase (HRP) and its corresponding derivatives indicates that histidine is not present in the first coordination sphere of heme in methemalbumin and that the protein is devoid of a well-defined heme cavity. The complex exhibits peroxidase activity by catalyzing oxidation of 2,2'-azinobis(3-ethylbenzthiazoline-6-sulfonate) by hydrogen peroxide. Its activity ( K(M)=433 microM, molar catalytic activity=0.33 s(-1)), however, is considerably lower compared to HRP, indicating differences in the heme environments. Fluorescence intensity decays of Trp214 in HSA and methemalbumin, best fitted to a three-exponential model, gave the lifetimes 7.03 ns (30%), 3.17 ns (38%), and 0.68 ns (32%) for HSA and 8.04 ns (1.7%), 2.42 ns (19.7%), and 0.64 ns (78.6%) for methemalbumin. These lifetime values were further confirmed by a model-independent maximum entropy method. Similarity in the lifetimes and variations in the amplitudes suggest that while conformational heterogeneity of HSA is unperturbed on heme binding, redistribution of the populations of the three conformations occurs and the sub-state associated with the shortest lifetime dominates the total population by approximately 80%. Decay associated spectra (DAS) indicate that the observed lifetime variation with wavelength is predominantly due to ground state heterogeneity, though solvent dipolar relaxation also contributes. Time-resolved fluorescence anisotropy measurements of the Trp214 residue yielded information on motion within the protein together with the whole protein molecule. The binding of heme did not affect the rotational correlation time of the albumin molecule (approximately 20 ns). However, the motion of tryptophan within the protein matrix increased by a factor of approximately 3 (0.46 ns to 0.15 ns). This indicates that while the overall hydrodynamic volume of the albumin molecule remained the same, tryptophan underwent a more rapid internal rotation because of the efficient energy transfer to the bound heme. Optical studies, analysis of lifetime measurements, DAS, and anisotropy measurements together suggest that heme binds to a surface residue. The rapid internal motion of Trp214 during its excited state lifetime for the approximately 80% populated conformer of methemalbumin allows the orientation factor, kappa(2), to approach the average value of 2/3. From the time-resolved fluorescence measurements and the energy transfer calculations on methemalbumin, a Trp214-heme distance of 22 A was deduced. PMID- 11935353 TI - An iron hydroxide moiety in the 1.35 A resolution structure of hydrogen peroxide derived myoglobin compound II at pH 5.2. AB - The biological conversions of O(2) and peroxides to water as well as certain incorporations of oxygen atoms into small organic molecules can be catalyzed by metal ions in different clusters or cofactors. The catalytic cycle of these reactions passes through similar metal-based complexes in which one oxygen- or peroxide-derived oxygen atom is coordinated to an oxidized form of the catalytic metal center. In haem-based peroxidases or oxygenases the ferryl (Fe(IV)O) form is important in compound I and compound II, which are two and one oxidation equivalents higher than the ferric (Fe(III)) form, respectively. In this study we report the 1.35 A structure of a compound II model protein, obtained by reacting hydrogen peroxide with ferric myoglobin at pH 5.2. The molecular geometry is virtually unchanged compared to the ferric form, indicating that these reactive intermediates do not undergo large structural changes. It is further suggested that at low pH the dominating compound II resonance form is a hydroxyl radical ferric iron rather than an oxo-ferryl form, based on the short hydrogen bonding to the distal histidine (2.70 A) and the Fe...O distance. The 1.92 A Fe...O distance is in agreement with an EXAFS study of compound II in horseradish peroxidase. PMID- 11935354 TI - Molecular modeling of the three-dimensional structure of Fe(II)-bleomycin: are the Co(II) and Fe(II) adducts isostructural? AB - The molecular modeling of Co(II)-bleomycin previously performed by us through NMR and molecular dynamics indicates that the most favorable structure for this complex is six-coordinate, with the secondary amine in beta-aminoalanine, the N5 and N1 nitrogens in the pyrimidine and imidazole rings, respectively, and the amide nitrogen in beta-hydroxyhistidine as equatorial ligands. The primary amine and either the carbamoyl group or a solvent molecule are proposed to occupy the axial sites. In this report, the results of the molecular modeling of Fe(II) bleomycin are presented. The NMR data for the ferrous derivative of the drug have already been reported by us, and were used here to generate the necessary restraints for this modeling work. For Co(II)-bleomycin, two new models exhibiting N-carbamoyl ligation to the metal centers were also assayed and compared with the ones previously examined. The results of this investigation on Fe(II)- and Co(II)-bleomycin are most consistent with a six-coordinate structure with five endogenous N-donors and a solvent molecule or the carbamoyl group as the sixth ligand. Comparisons of the best Co(II)- and Fe(II)-bleomycin models with the NMR-generated structures for some relevant metallo-BLMs favor the model with only endogenous ligands and N-carbamoyl ligation as the structure probably held in solution by both Co(II)- and Fe(II)-bleomycin. PMID- 11935355 TI - Interactions in solution of cobalt(II) and nickel(II) with nicotinamide adenine dinucleotide: a potentiometric and calorimetric study. AB - The equilibrium constants and the thermodynamic parameters enthalpy and entropy of the interaction between Ni(II) and Co(II) with NAD(+) in aqueous solution were determined by calorimetry and potentiometry methods (ionic strength adjusted to 0.1 with sodium nitrate at 25 degrees C). The macrochelation of the systems was also studied. All the data, including the protonation enthalpy data of NAD(+) (very similar to the protonation enthalpy of 5'-AMP) suggest a less restrictive model for the NAD(+) "folded" conformation without intramolecular stacking between the bases, in agreement both with recent theoretical calculations and with the X-ray structure of trimethylene-bisadeninium or the free acid form of NAD(+). PMID- 11935356 TI - IR spectroelectrochemical study of the binding of carbon monoxide to the active site of Desulfovibrio fructosovorans Ni-Fe hydrogenase. AB - The binding of carbon monoxide, a competitive inhibitor of many hydrogenases, to the active site of Desulfovibrio fructosovorans hydrogenase has been studied by infrared spectroscopy in a spectroelectrochemical cell. Direct evidence has been obtained of which redox states of the enzyme can bind extrinsic CO. Redox states A, B and SU do not bind extrinsic CO; only after reductive activation of the hydrogenase can CO bind to the active site. Two states with bound extrinsic CO can be distinguished by FTIR. These two states are in redox equilibrium and are most probably due to different oxidation states of the proximal 4Fe-4S cluster. Vibrational frequencies and theoretical quantum mechanics studies (DFT) of this process preclude the possibility of strong bonding of extrinsic CO to the Fe or Ni atoms of the active site. We propose that CO inhibition is caused by weak interaction of the extrinsic ligand with the Ni atom, blocking electron and proton transfer at the active site. A calculated structure with a weakly bound extrinsic CO at Ni has relative CO frequencies in excellent agreement with the experimental ones. PMID- 11935357 TI - Thermodynamic and spectroscopic study of Cu(II) and Ni(II) binding to bovine serum albumin. AB - The thermodynamics of Cu(II) and Ni(II) binding to bovine serum albumin (BSA) have been studied by isothermal titration calorimetry (ITC). The Cu(II) binding affinity of the N-terminal protein site is quantitatively higher when the single free thiol, Cys-34, is reduced (mercaptalbumin), compared to when it is oxidized or derivatized with N-ethylmaleimide. This increased affinity is due predominantly to entropic factors. At higher pH (approximately 9), when the protein is in the basic (B) form, a second Cu(II) binds with high affinity to albumin with reduced Cys-34. The Cu(II) coordination has been characterized by UV vis absorption, CD, and EPR spectroscopy, and the spectral data are consistent with thiolate coordination to a tetragonal Cu(II), indicating this is a type 2 copper site with thiolate ligation. Nickel(II) binding to the N-terminal site of BSA is also modulated by the redox/ligation state of Cys-34, with higher Ni(II) affinity for mercaptalbumin, the predominant circulating form of the protein. PMID- 11935358 TI - Pulsed ELDOR spectroscopy of the Mo(V)/Fe(III) state of sulfite oxidase prepared by one-electron reduction with Ti(III) citrate. AB - The titration of chicken liver sulfite oxidase (SO) with the one-electron reductant Ti(III) citrate, at pH 7.0, results in nearly quantitative selective reduction of the Mo(VI) center to Mo(V), while the b-type heme center remains in the fully oxidized Fe(III) state. The selective reduction of the Mo(VI/V) couple has been established from electronic and EPR spectra. The electronic spectrum of the Fe(III) heme center is essentially unchanged during the titration, and the continuous wave (CW)-EPR spectrum shows the appearance of the well-known Mo(V) signal due to the low pH ( lpH) form of SO. Further confirmation of the selective formation of the Mo(V)/Fe(III) form of SO is provided by the approximately 1:1 ratio of the integrated intensities of the Mo(V) and low-spin Fe(III) EPR signals after addition of one equivalent of Ti(III). The selective generation of the Mo(V)/Fe(III) form of SO is unexpected, considering that previous microcoulometry and flash photolysis investigations have indicated that the Mo(VI/V) and Fe(III/II) couples of SO have similar reduction potentials at pH 7. The nearly quantitative preparation of the one-electron reduced Mo(V)/Fe(III) form of SO by reduction with Ti(III) has enabled the interaction between these two paramagnetic metal centers, which are linked by a flexible loop with no secondary structure, to be investigated for the first time by variable-frequency pulsed electron electron double resonance (ELDOR) spectroscopy. The ELDOR kinetics were obtained from frozen solutions at 4.2 K at several microwave frequencies by pumping on the narrow Mo(V) signal and observing the effect on the Fe(III) primary echo at both higher and lower frequencies within the microwave C-band region. The ELDOR data indicate that freezing the solution of one-electron reduced SO produces localized regions where the concentration of SO approaches that in the crystal structure, which results in the interpair interactions being the dominant dipolar interaction. However, thorough analysis of the ELDOR decay curves and simulations suggests a distribution of intramolecular Mo...Fe distances, consistent with the proposal of multiple conformations in solution for the flexible loop that connects the Mo and heme domains of SO. PMID- 11935363 TI - Overview of mathematical computer models of striated sphincter muscles. AB - The present paper compares current mathematical striated sphincter models. Current models are subdivided in four categories: (1) simple models, (2) implementations of the urethral resistance relation, (3) models with realistic muscle dynamics and (4) finite element models. In our research group a neural network model, representing Onuf's nucleus, the spinal motor nucleus that innervates the external urethral and anal sphincters, was developed. A realistic sphincter model is needed to test the neural network. To decide whether or not a model is applicable in our research two requirements should be fulfilled: (1) the presence of realistic muscle dynamics preferably by implementation of a Huxley type muscle model and (2) the model should consist of more than one muscle unit to form a more dimensional model. Reviewing the literature, if a myogenic sphincter is modelled, mainly the Hill-equation is applied. Moreover, single muscle unit models are published. In general a multi-unit muscle model of the sphincter is lacking, prohibiting the study of the inherent properties of sphincter muscles, which could give information on the realistic behaviour of elements in circular muscles. It is concluded that the functionality of current sphincter models is limited for our purpose. PMID- 11935364 TI - Electrical stimulation of the trigeminal tract in chronic, intractable facial neuralgia. AB - In this paper the treatment of patients with chronic, intractable trigeminal neuralgia by invasive electrical stimulation of the Gasserion ganglion is reviewed. Two different surgical techniques are employed in this treatment. Most frequently, a method similar to the traditional technique for percutaneous glycerol and radiofrequency trigeminal rhizolysis is used: a small percutaneous stimulation electrode is advanced under fluoroscopic control through a thin needle via the foramen ovale to the Gasserian cistern. Some neurosurgeons use an open surgical technique by which the Gasserian ganglion is approached subtemporally and extradurally, and the bipolar pad electrode is sutured to the dura. When percutaneous test stimulation is successful (at least 50% pain relief) the electrode is internalized and connected to a subcutaneous pulse generator or RF-receiver. Data from 8 clinical studies, including 267 patients have been reviewed. Of all 233 patients with medication-resistant atypical trigeminal neuralgia 48% had at least 50% long term pain relief. The result of test stimulation is a good predictor of the long term effect, because 83% of all patients with successful test stimulation had at least 50% long term relief, and 70% had at least 75% long term relief. Patients generally preferred this invasive method over TENS. The success rate in patients with postherpetic trigeminal neuralgia was very low (less than 10%). It is suggested that the likelihood of pain relief by electrical stimulation is inversely related to the degree of sensory loss. It is concluded that invasive stimulation of the Gasserian ganglion is a promising treatment modality for patients with chronic, intractable, atypical trigeminal neuralgia. PMID- 11935365 TI - Excessive reflexes in spinal cord injury triggered by electrical stimulation. AB - Interaction of electrocutaneous stimulation with an impaired human motor control system may result in unstable reflex loops causing excessive spastic reactions. These contractions are usually excluded from analysis since the presence of spasm is one of the criteria commonly applied for discarding a contraction. They may, however, provide interesting information on the nature of spasticity. The dorsiflexor muscles of four SCI subjects were activated by means of surface electrical stimulation and the isometric ankle moment was measured. Short bursts of constant stimulation frequency at seven different frequencies (8, 12, 16, 20, 25, 33, 50 Hz) triggered spastic reactions in all subjects. The onset times of spastic activity during an electrically elicited contraction shortened with increased stimulation frequency. A stimulation burst may also have a spasticity reduction effect on a subsequent burst, indicating potential short term therapeutic effects of stimulation on spasticity in isometric conditions. PMID- 11935366 TI - A model for regulation of the Mg(2+)-stimulated acto-myosin-ATPase activity: inhibition of the formation of actin-myosin complex and the Mg( 2+)-stimulated acto-myosin-ATPase activity by IMP and AMP. AB - Previously, we showed that the decrease in force output during continuous isometric contractions in rat skeletal muscle was related to an increase in the concentration of IMP. In this paper we report on additional experiments in which the effect of IMP on the Mg(2+)-stimulated acto-myosin-ATPase activity of isolated actin and myosin is measured at 35 degrees C. The results show that 1) the binding of actin to myosin is co-operative (Hill coefficient = 3.82); 2) in the presence of IMP or AMP the Mg(2+)-stimulated acto-myosin-ATPase activity is inhibited up to 60% at 10 mM; 3) in the presence of IMP or AMP not only the Mg(2+)-stimulated acto-myosin-ATPase activity decreases, but also K(50). From these results we conclude that IMP and AMP may be considered as uncompetitive inhibitors. Our results suggest that IMP and AMP can prevent an 'energy crisis' during exhaustive exercise of short duration by down-regulating the contractile machinery. PMID- 11935367 TI - Oxidative modification of rat liver 5'-nucleotidase: the mechanisms for protection and re-activation. AB - The effect of oxidative stress catalysed by transition metals appears to have a critical relevance for the structure and function not only of membrane lipids but also of integral membrane proteins in a complex lipid-protein assembling, and membrane-dependent function. The integral membrane enzyme 5'-nucleotidase is susceptible to Fe((2+))-ion catalysed oxidative modification, and the extent of enzyme inhibition is in inverse relationship (r = -0.820) with lipid peroxidation (MDA) level. This work is also a comparative study about possible effectiveness of different Fe-ion chelators (deferoxamine, Na-citrate, Na-salicylate, ammonium oxalate and EDTA), antioxidants (GSH, GSH/GSH-Px system, Cu, Zn-SOD and mannitol) and metal cations (Mg(2+) and Mn(2+)) to protect or restore Fe(2+)-ion induced 5' nucleotidase inhibition and to suppress Fe(2+)-ion enhanced lipid peroxidation. Among the examined chelators it was only deferoxamine and Na-citrate that exerted a fully protective and reactivating ability; among the antioxidants it was only GSH; among the metal cations it was only Mn(2+). The ability to protect or restore 5'-nucleotidase activity and to diminish chain-induced lipid peroxidation is explicable in terms of: metal-binding ability, capacity of taking iron away from a biological molecule, or ability of transferring the damage to itself. After a short incubation period, the iron associated with enzyme or lipid hydroperoxides could be in a labile coordinative linkage, still able to interact with possible ligands or metal cations. PMID- 11935368 TI - Evaluation of oxidative stress in patients with cancer. AB - The oxidative stress is considered to be involved in the pathophysiology of cancers. In the current study we explored the oxidative stress in patients with different cancers and corresponding benign diseases by evaluation of the level of lipid peroxidation products (MDA level) in the plasma and the activity of erythrocyte antioxidant defense enzymes superoxide dismutase (SOD) and catalase (CAT). Significantly higher plasma levels of lipid peroxidation products were detected in patients with early and advanced cancers in comparison to the healthy volunteers (mean 3.1 micromol/l and 2.3 micromol/l, p = 0.0003 and p = 0.029, respectively, t-test). In addition, 10-20 days after radical operations of cancer patients with normal postoperative recovery period, the plasma levels of MDA decreased and reached values close to the controls (mean 2.0 micromol/l). SOD in erythrocytes of patients with benign diseases and malignant solid tumors before and after surgery did not differ from that of the controls. In contrast, CAT activity of patients with early cancers was found to be significant higher than that of the controls (mean 22157.2 U/gHb vs. 12832.0 U/gHb, p = 0.032, t-test). A decrease of CAT activity was observed after surgery (mean 15225.0 U/gHb). In conclusion, our results suggest the presence of an increased oxidative stress accompanied by a lack of changes of erythrocyte SOD activity and an adaptive increase of CAT activity. PMID- 11935369 TI - Study of thyroid gene expression and regulation by DNA array technology: pitfalls and necessary controls in the use of commercial filters. AB - In order to identify the genes differentially expressed by TSH stimulation in dog and human thyrocytes we have used the gene array technology. Some modifications of the standard procedure were introduced to improve the method. While the macroarray technology has been developed to identify genes differentially expressed between two tissues, we report here that certain widely publicized commercial nylon filters do not allow to achieve this result. Because each step is crucial to the method and because it relies on quantitative measurements, we report different pitfalls of the method when commercial nylon filters with spotted bacterial colonies are used. The list of the most expressed genes in dog and human thyrocytes, which constitutes the minimal transcriptome of these tissues, has nevertheless been drawn up. Control tests that should be applied before the experimental use of the very expensive arrays are proposed. PMID- 11935370 TI - Upregulation of the transport system for TNFalpha at the blood-brain barrier. AB - The transport system for the cytokine tumor necrosis factor-alpha (TNFalpha) at the blood-brain barrier (BBB) enables an enhanced yet saturable entry of TNFalpha from blood to the CNS. This review focuses on the selective upregulation of the transport system for TNFalpha at the BBB that is specific for type of pathology, region, and time. The upregulation is reflected by increased CNS tissue uptake of radiolabeled TNFalpha after iv injection in mice and by inhibition of this increase with excess non-radiolabeled TNFalpha. (1) Spinal cord injury (SCI): upregulation of TNFalpha uptake after thoracic transection is seen in the delayed phase of BBB disruption at the lumbar spinal cord. Thoracic SCI by compression, however, has a longer lasting impact on TNFalpha transport that involves thoracic and lumbar spinal cord, in contrast to the upregulation confined to the lumbar region in lumbar SCI by compression. Regardless, the uptake of TNFalpha by spinal cord does not parallel BBB disruption as measured by the leakage of radiolabeled albumin. (2) Experimental autoimmune encephalomyelitis (EAE): the increase in the differential permeability to TNFalpha is seen in all CNS regions (brain and cervical, thoracic, and lumbar spinal cord) and has a distinct time course and reversibility. Exogenous TNFalpha has biphasic effects in modulating functional scores. The BBB, a dynamically regulated barrier, is actively involved in disease processes. PMID- 11935371 TI - Nerve growth factor and neuroimmune responses: basic and clinical observations. AB - Nerve growth factor (NGF) is a polypeptide that in addition to its effect on survival of peripheral and brain neuron acts also on a variety of cells localized in the immune system. We have recently shown that the constitutive levels of NGF undergo trough significant changes during inflammatory conditions and in neuroimmune pathologies. We have also reported that autoimmune inflammatory disorders are characterized by a disregulation of synthesis and utilization of NGF. Though the mechanisms implicated in these events are not yet known, there emerging evidence indicating this polypeptide can play an important role in healing processes within the nervous and cutaneous tissues and in inflammatory conditions. PMID- 11935372 TI - NGF, BDNF, leptin, and mast cells in human coronary atherosclerosis and metabolic syndrome. AB - While multiple growth factor, cytokines, and immune cells are identified in atherosclerotic lesions, as well as an essential nonneuronal function of neurotrophins implicated in cardiovascular tissue development and in lipid and glucose metabolism, the role of the neurotrophins NGF and BDNF and also the adipokine leptin in human coronary atherosclerosis and related disorders, such as metabolic syndrome, remains unclear. Here we report that (i) both the amount and the immunoreactivity of NGF was reduced and the expression of p75NGF receptor and the number of mast cell increased in human atherosclerotic coronary arteries (n = 12) compared with control specimens (n = 9) obtained from autopsy cases, and (ii) NGF and BDNF plasma levels were reduced in patients with metabolic syndrome (n = 23) compared with control subjects (n = 10). Also, in metabolic syndrome patients, a positive correlation between the plasma leptin levels and the number of adipose tissue mast cells was found, suggesting that leptin may be a novel adipoimmune mediator. Altogether, the results provide the first correlative evidence for the potential involvement of NGF, BDNF, leptin, and mast cells in human coronary atherosclerosis and metabolic syndrome, implying neuroimmune and adipoimmune pathways in the pathobiology of these cardiovascular disorders. PMID- 11935373 TI - Neuroinflammatory implication of Schistosoma mansoni infection in the mouse. AB - Schistosomiasis is a parasitic disease due to Schistosoma mansoni. Schistosome infection is known to induce granulomas not only in the spleen, bladder, liver and intestine but also in the brain and spinal cord resulting in severe neuropathological and psychiatric disorders though the interaction between Schistosoma mansoni infection and the nervous system has received on the whole little attention. In the present review it has been discussed recent findings from experimental Schistosoma mansoni infection in mouse nervous system. We show that brain granulomas are associated with a significant alteration in the constitutive levels of nerve growth factor (NGF), a trophic factor playing an essential role in nerve growth and differentiation and in preventing neuronal damages. Animals infected with schistosomes suffered also of increased pain sensitivity which was inhibited by TNF-alpha antibody injections and not by anti NGF. These findings suggest that the neuropathological dysfunctions in neuroschistosomiasis may be linked to changes in the basal levels and/or activity of neurotrophic factors caused by local formation of granulomas. PMID- 11935374 TI - TH2 lymphocytes secrete functional VIP upon antigen stimulation. AB - In addition to the peptidergic innervation, immune cells may also represent a source for VIP in the lymphoid organs. Previous studies reported increased VIP mRNA and protein expression in mitogen-stimulated B and T lymphocytes. To determine whether specific T cell subsets are responsible for VIP production, we derived TH1 and TH2 effector cell lines from T-cell receptor transgenic mice. TH1 and TH2 cells were stimulated with the specific (pigeon cytochrome C peptide) or nonspecific (ovalbumin) antigen presented by MHC class II compatible antigen presenting cells. Upon stimulation with the specific antigen, TH2, but not TH1 cells express VIP mRNA and intracelllular VIP protein, as determined by Northern blots and FACS analysis. Supernatants harvested from antigen-stimulated TH2 cells contain secreted VIP, as determined by Elisa, and induce cAMP in HEK293 cells transfected with the specific VIP/PACAP receptor VPAC1. These results confirm that TH2, but not TH1 cells, express and secrete functional VIP following specific antigen stimulation. The release of VIP within the lymphoid microenvironment following antigenic stimulation provides a physiological basis for the immunoregulatory effects of VIP on neighboring immune cells, such as downregulation of macrophage activation, effects on lymphocyte migration, on antigen-induced T cell apoptosis, and on T cell differentiation. PMID- 11935375 TI - Future aspects of psychoneuroimmunology - lymphocyte peptides reflecting psychiatric disorders studied by mass spectrometry. AB - We have investigated whether cytoplasmatic and nuclear extracts of human peripheral blood lymphocytes contain arginine vasopressin (AVP), of importance for memory functions, in samples from healthy controls and patients diagnosed as depressed. It is the first time as AVP, AVP-fragments and chemically modified AVP forms have been demonstrated in lymphocyte/nuclear extracts. This was performed by an HPLC-purification step, followed by a second immunoprecipitation step before identification by mass spectrometry. We are developing new methods using a combination of high-resolution mass spectrometry and separation techniques such as capillary electrophoresis and nano liquid chromatography. We have named this methodological approach when studying endogenous peptides -Peptidomics. PMID- 11935376 TI - VIP and PACAP enhance the in vivo generation of memory TH2 cells by inhibiting peripheral deletion of antigen-specific effectors. AB - In an immune response, antigen-specific CD4 T cells proliferate and differentiate into effector cells capable to produce large amounts of cytokines upon restimulation. Most effector T cells are later eliminated through antigen-induced cell death (AICD), mediated through FasL/Fas interactions. A low percentage of effector T cells survive and differentiate into long-lived memory cells. Mechanisms must operate not only to destroy no longer needed and even potentially damaging T cells, but also to allow the survival of a small number of activated T cells. Little is known about the factors and mechanisms that regulate the shift from an apoptosis-sensitive to an apoptosis-resistant phenotype. VIP and the structurally related peptide, PACAP, synthesized and/or released in the immune organs act on both innate and adaptive immunity. Recently, VIP and PACAP were shown to inhibit AICD in peripheral CD4 T cells by down-regulating FasL expression. In view of these findings, VIP and PACAP are reasonable candidates for the generation of memory T cells. To test this hypothesis, we analyzed the effects of VIP and PACAP in various models for effector and memory T cells. Our data demonstrate that both neuropeptides promote the in vivo effector function and memory phenotype of Th2, but not Th1 cells, by preferentially inhibiting the clonal deletion of Th2 cells. To our knowledge, this is the first report describing the role of a neuropeptide present in the lymphoid microenvironment on the generation and maintenance of long-lived memory T cells. PMID- 11935377 TI - Inhibition of endotoxin-induced macrophage chemokine production by VIP and PACAP in vitro and in vivo. AB - Inflammatory chemokines recruit immune cells which initiate and maintain the inflammatory response. Although such a response is necessary for the elimination of the antigen, the inflammation has to be eventually resolved. Peptides such as vasoactive intestinal peptide (VIP) and pituitary adenylate cyclase-activating polypeptide (PACAP), released following antigenic stimulation, contribute to the termination of an inflammatory response primarily by inhibiting the production of proinflammatory cytokines. Here we investigated the effects of VIP and PACAP on chemokine production. We report that VIP and PACAP inhibit the expression of the macrophage-derived CXC chemokines MIP-2 and KC (IL-8), and of the CC chemokines MIP-1a, MIP-1b, MCP-1 and RANTES in vivo and in vitro. The decrease of chemokine gene expression correlates with an inhibitory effect of VIP/PACAP on NFkB binding. In an in vivo model of acute peritonitis, the inhibition of chemokine production by VIP/PACAP leads to a significant reduction in the recruitment of PMNs, macrophages and lymphocytes into the peritoneal cavity. These findings support the proposed role of VIP and PACAP as key endogenous anti-inflammatory agents, and describe a novel mechanism, i.e., the inhibition of the production of macrophage-derived chemokines. PMID- 11935378 TI - Impact of stress, gender and menstrual cycle on immune system: possible role of nitric oxide. AB - Stress is a factor found to be involved in the etiology of many diseases. Gender and menstrual cycle phases are other factors affecting the predisposition of individuals for certain diseases. Results from animal and human studies suggest that the distribution of immune system cells may change at different phases of the menstrual cycle. Acute mental stress in humans alters immune variables, too. The increase in the number of natural killer (NK) cells is the most consistent finding among the immune variables, though there are controversies for the other lymphocyte groups. Nitric oxide (NO) as an immune mediator has an unsettled role whether it causes the redistribution of the immune cells, or is an end product of lymphocyte activation. This study was planned to investigate the effect of mental stress on lymphocyte subtypes and the role of NO, for men and women at different phases of the cycle. For this purpose, healthy men (n = 10) and women (n = 10), during the follicular and luteal phases underwent Stroop colour-word interference and cold pressor tests. The immune system responses before and after the tests were determined by cell counts with the flowcytometer. Menstrual cycle phase was ascertained by plasma estrogen and progesterone measurements. Stress response was evaluated by blood pressure (BP) and heart rate (HR) measurements throughout the tests and plasma cortisol and urinary metanephrine and vanillylmandelic acid (VMA) measurements before and after the tests. Plasma and urinary NO determinations were performed before and after the test was completed. All the results were analysed with the appropriate statistical methods. The luteal phase differed from the other groups due to the presence of suppressed immune response to acute stress, including decreased CD4/CD8 ratio and NK cell percentage. On the other hand, acute stress caused a shift from cellular to humoral immunity in men. As indicated by these results, individual reaction towards stress is affected by gender and menstrual cycle phase. NO appears to be a possible effector molecule for these differences. PMID- 11935379 TI - Demonstration of spontaneous and stretch induced urinary bladder EMG in the living rabbit. AB - Spontaneous bladder EMG was recorded in the living rabbit from an isovolumetric bladder without chemical or electrical stimulation. Mechanical intervention, either by lifting the bladder out of the abdomen or by rapid filling, resulted in stretch induced bladder EMG. A self made epoxy resin electrode device that embedded 32 EMG recording electrodes in a matrix like pattern, each electrode Ag/AgCl, d = 0.6 mm with an interdistance of 2.3 mm, was used for registration. The recorder used a common average reference technique and a sample frequency of 400 Hz. A signal bandwidth of 0.05 to 108 Hz was available for analysis. Spontaneous EMG consisted of single spikes and bursts (2-20 spikes), but not of continuous activity. The shape of spikes was triphasic. Single spikes appeared with and without burst activity. Small (2-5 spikes) and large bursts (6-20 spikes) were discerned; small bursts not necessarily propagated across electrodes, large bursts did and were able to organize, suggesting that they were under short neuron system control. Spontaneous EMG was probably related to both contraction and relaxation. Stretch induced EMG was characterised by continuous activity on all electrodes, spikes that followed each other immediately, slowly fading away. The spikes had an elongated third phase when compared to the shape of spontaneous activity. Highest activity and amplitudes were found after lifting the bladder out of the abdomen and placing it on the electrode device. A concept is put forward in which the continuous activity is not unequivocally related to muscle shortening, but where the current stress and strain situation of the bladder tissue can cause a muscle fibre elongation upon the appearance of electrical activity. The EMG activity found was in many aspects similar to results of a previous study using mortalized rabbits. Artifact sources like the heart, respiration, or local movement between electrode and bladder could easily be identified due to the new experimental methodology used. PMID- 11935380 TI - Angiotensin II-induced growth effects in vascular smooth muscle in cell culture and in the aortic tunica media in organ culture. AB - Several different studies have investigated the growth effects of angiotensin II on vascular smooth muscle cells in culture. However, smooth muscle cells change their phenotype when placed in culture. The objective of the present study was to investigate the effects of angiotensin II on (3)H-thymidine and (3)H-proline incorporation in vascular smooth muscle cells in culture and in the tunica media of blood vessels perfused at normal physiological pressures in organ culture, thus avoiding the phenotypic changes observed in cell culture. The perfusion system consisted of a peristaltic pump and a closed circuit of plastic tubing connected to a culture media bottle where thoracic rat aortae were placed. Angiotensin II induced an increase in (3)H-thymidine and (3)H-proline incorporation in both culture systems. The results suggest that angiotensin II may play a role in mediating cell growth in vascular smooth muscle cells in their 'contractile' as well as in their 'synthetic' phenotype. PMID- 11935381 TI - Twitch and tetanic tension during culture of mature Xenopus laevis single muscle fibres. AB - Investigation of the mechanisms of muscle adaptation requires independent control of the regulating factors. The aim of the present study was to develop a serum free medium to culture mature single muscle fibres of Xenopus laevis. As an example, we used the culture system to study adaptation of twitch and tetanic force characteristics, number of sarcomeres in series and fibre cross-section. Fibres dissected from m. iliofibularis (n = 10) were kept in culture at a fibre mean sarcomere length of 2.3 microm in a culture medium without serum. Twitch and tetanic tension were determined daily. Before and after culture the number of sarcomeres was determined by laser diffraction and fibre cross-sectional area (CSA) was determined by microscopy. For five fibres twitch tension increased during culture and tetanic tension was stable for periods varying from 8 to 14 days ('stable fibres'), after which fibres were removed from culture for analysis. Fibre CSA and the number of sarcomeres in series remained constant during culture. Five other fibres showed a substantial reduction in twitch and tetanic tension within the first five days of culture ('unstable fibres'). After 7-9 days of culture, three of these fibres died. For two of the unstable fibres, after the substantial force reduction, twitch and tetanic tension increased again. Finally at day 14 and 18 of culture, respectively, the tensions attained values higher than their original values. For stable fibres, twitch contraction time, twitch half-relaxation time and tetanus 10%-relaxation time increased during culture. For unstable fibres these parameters fluctuated. For all fibres the stimulus threshold fluctuated during the first two days, and then remained constant, even for the fibres that were cultured for at least two weeks. It is concluded that the present culture system for mature muscle fibres allows long term studies within a well-defined medium. Unfortunately, initial tetanic and twitch force are poor predictors of the long-term behaviour of the fibres. PMID- 11935382 TI - Study of the mechanism of rectal motility: the 'mass squeeze contraction'. AB - The motor physiology of the rectum has remained largely obscure, especially concerning the mechanism of rectal motility. In the current communication we tested the possibility of characterizing the mechanism of rectal motility during filling and evacuation through the study of the rectal electric activity in 16 healthy volunteers (mean age 43.6 +/- 10.8 years; 11 men). Two monopolar silver silver chloride electrodes were introduced per annum and fixed to the rectal mucosa by suction. The rectum was distended in 10 ml increments of water by means of a balloon-ended catheter inserted into the rectum. The rectal pressure was measured by one catheter placed above and a second one below the rectal balloon, and the 2 catheters were connected to 2 strain gauge pressure transducers. Regular triphasic slow waves or pacesetter potentials (PPs) were recorded from the 2 electrodes at rest. PPs were superimposed or followed randomly by action potentials (APs). APs but not PPs were coupled with elevated rectal pressure. Rectal distension with 10 ml of water caused no significant changes of the rectal pressure or EMG activity. Distension with a mean volume of 27.3 +/- 4.7 ml effected a significant increase (p < 0.05) of the rectal electromechanical activity proximally to the balloon and a decrease distally (p < 0.05) to it. With progressive increase of the rectal distension, the electromechanical activity continued to increase proximally and to decrease distally to the balloon, until, at a mean distending volume of 76.3 +/- 3.7 ml, the balloon was dispelled to the exterior. In conclusion, the identification of the modality of rectal motility during defecation was feasible by recording the rectal electromechanical activity. The rectal contraction is suggested to occur in a 'mass squeeze manner' which squeezes the rectal contents aborally into the anal canal. The recognition of the rectal motor modality appears to be important for the understanding of rectal motility disorders. However, further studies are required to confirm these findings. PMID- 11935383 TI - Evaluating the time limit at maximum aerobic speed in elite swimmers. Training implications. AB - The aim of the present study was to make use of the concepts of maximum aerobic speed (MAS) and time limit (tlim) in order to determine the relationship between these two elements, and this in an attempt to significantly improve both speed and swimming performance during a training season. To this same end, an intermittent training model was used, which was adapted to the value obtained for the time limit at maximum aerobic speed. During a 12 week training period, the maximum aerobic speed for a group of 9 top-ranking varsity swimmers was measured on two occasions, as was the tlim. The values generated indicated that: 1) there was an inverse relationship between MAS and the time this speed could be maintained, thus confirming the studies by Billat et al. (1994b); 2) a significant increase in MAS occurred over the 12 week period, although no such evolution was seen for the tlim; 3) there was an improvement in results; 4) the time limit could be used in designing a training program based on intermittent exercises. In addition, results of the present study should allow swimming coaches to draw up individualized training programs for a given swimmer by taking into consideration maximum aerobic speed, time limit and propelling efficiency. PMID- 11935384 TI - Effect of varying light intensity on maximal power production and selected metabolic variables. AB - This study was designed to examine the effect of exposure to two levels of light intensity (bright; 5000 lux, dim; 50 lux) prior to supramaximal cycle exercise on performance and metabolic alterations. The exercise was performed after bright and dim light exposure for 90 minutes. Ten male long-distance runners volunteered to take part in the study. They performed 45-sec supramaximal exercise using a cycle ergometer in a 500-lux. Mean power output was measured during the exercise. Lactate and ammonia in the blood and epinephrine and norepinephrine concentrations in plasma were measured at rest immediately after bright and dim light exposures and after the exercise. Bright and dim light exposure prior to exercise did not significantly affect the power output during the exercise. Blood glucose concentration immediately after exercise and plasma epinephrine during the resting period were significantly lower after bright light exposure compared with dim light exposure (p < 0.05). No significant difference was found in blood lactate, ammonia, or plasma norepinephrine levels after exercise following bright and dim light exposures. This study demonstrated that bright light stimulation prior to supramaximal exercise decreases glucose and epinephrine levels, but is not related to physical performance. PMID- 11935386 TI - Purification and properties of insulin-activated nitric oxide synthase from human erythrocyte membranes. AB - A membrane bound form of nitric oxide synthase of human erythrocytes that could be activated by insulin was purified to homogeneity by detergent solubilization of the purified membrane preparation of these cells. The purified enzyme (M(r) 230 KD) was found to be composed of one heavy chain (M(r) 135 KD) and one light chain (Mr 95 KD) held together by disulphide bond(s). Scatchard plot analysis of insulin binding to the purified enzyme showed the presence of 2 different populations of the binding sites and the activation were directly related to the hormone binding to the protein. Line weaver Burk plot of the purified enzyme showed that the stimulation of the enzymic activity by insulin was related to the decrease of K(m) with simultaneous increase of V(max). Treatment of the purified enzyme with anti insulin receptor antibody inhibited the activation of the enzyme and the binding of the hormone to the protein. Furthermore NO itself, at low concentration (<0.4 microM) activated the enzyme, but at higher concentration (>0.8 microM) had no effect on the activation. Incubation of the purified enzyme with insulin simultaneously stimulated the tyrosine kinase and nitric oxide synthase activities of the preparations, that could be inhibited by genistein (an inhibitor of tyrosine kinase). These results indicated that the insulin activated nitric oxide synthase could be the insulin receptor itself. PMID- 11935385 TI - Simvastatin effects on a human lung carcinoma and cholesterol homeostasis of host and non-host mice. AB - In order to investigate the effect of a competitive inhibitor of the HMG-CoA reductase on tumor growth and cholesterol homeostasis of host and non-host mice, we maintained a human lung mucoepidermoid carcinoma (HLMC) in nude mice, treating these animals with Simvastatin for 33 days. The drug increased the total activity of hepatic HMG-CoA reductase without affecting the cholesterolemia. Non-treated host animals presented lower serum, tissue and microsomal hepatic cholesterol than non-host animals. These differences disappeared when animals were treated with Simvastatin, though the induction of the reductase activity at mid-dark was higher in non-host than in host animals. Simvastatin produced no significant effects on both final tumor volume and body weight. Synthesis and cholesterol homeostasis restoration induced by liver and tumoral reductase would account for no effect on the HLMC growth after a long treatment with Simvastatin. PMID- 11935387 TI - Pregnancy in adolescent rats, growth and neurodevelopment in their offspring. AB - Pregnancy, lactation and the relationship between mother and their offspring in adolescence was studied in terms of pups neurosomatic development. The frequency of conception and birth rate were reduced in adolescent dams. Their body weight gain was accelerated during the first two weeks of pregnancy, while a significant delay occurred in the third week. At birth, plasma corticosterone level in the neonates was increased. Adolescent dams ingested less food during the first week of gestation. Following that, till the second week of lactation, the food consumption was equal to that of control group. Although adolescent pups were heavier at birth, the reduction of their number and of their body weight occurred during lactation. Judging by appearance of grasping, righting, placing and of negative geotaxis reflexes, the delay in their neurological maturation was also present. The reason for the growth and neurodevelopmental delay during lactation is probably the result of malnutrition and stress of disturbed mother - infant relationship in adolescent litters, which lasted at least during the two postnatal weeks. It was indicated by the resting plasma corticosterone levels during the first two postnatal weeks. This finding suggests that the pregnancy in adolescent rats induces delay in physical and neurological development, as well as in the increased rate of postnatal mortality of their offspring. PMID- 11935388 TI - Effect of chronic lead exposure on kidney function in male and female rats: determination of a lead exposure biomarker. AB - Several cytotoxic chemical pollutants inducing peroxidative damages are liable to induce kidney failure. Among these pollutants we find heavy metals such as: lead, nickel, cadmium, vanadium and mercury. Lead is one of the most dangerous metals because it is widely spread in the environment, and because it may be a source of several nervous diseases. The aim of this study is to provide evidence concerning the effect of this metal on the renal function and to try to determine a storage corner in the organism which serves as an indicator of a lead intoxication. Lead acetate was administered by oral route in the drinking water to adult rats aged three months at the rate of 0.3% (P1) and 0.6% (P2). Reference rats received distilled water to drink under the same conditions. The treatment continued for 15, 30, 45, 60 and 90 days. The creatinemia, uremia, glycemia and creatinuria are determined by colorimetric techniques. Lead concentration in blood as well as the lead content of the tail are determined by atomic absorption after nitroperchloric mineralization at the liquid stage. The results showed an increase of creatinemia on the 30th day of the experiment for both sexes in (P1 and P2). The same happened for ureamia. The increase of these two parameters would indicate a renal deficiency which is confirmed by a decrease of creatinuria and urinary pH observed mainly on and after the 45th day of the experiment. An increase of the renal relative weight was noticed in P1 and P2 on the 30th day of the treatment. The determination of the concentration of lead in the blood shows that this factor increases among treated subjects in a constant way, independently of the dose and the duration of the treatment. Nevertheless, the rate increase of lead in the tail seems to be dose-dependent. In conclusion, lead administered by oral route causes a renal deficiency to the rat without distinction between males and females. In addition, the tail seems to be a reliable exposure biomarker that demonstrates lead intoxication. The tail seems to be a dosimeter of lead bio-accumulation. It constitutes an endogenous source of lead impregnation. The concentration of lead in the blood is only an indicator of recent exposure. PMID- 11935393 TI - The anterior pituitary gland is mysterious, alluring and useful. PMID- 11935395 TI - Combined expression of different hormone genes in single cells of normal rat and mouse pituitary. AB - Cells displaying combined expression of different pituitary hormone genes (further referred to as 'multi-hormone mRNA cells') were identified in normal rat and mouse pituitary by single cell RT-PCR. These cells do not seem to produce or store all the respective hormones the mRNAs encode for. The cells are already developed at day 16 of embryonic life (E16) in the mouse. Different peptides, such as gamma3-melanocyte-stimulating hormone (gamma3-MSH) and gonadotropin releasing hormone (GnRH), affect different subsets of these cells. In culture, estrogen and GnRH increase the number of 'multi-hormone mRNA cells' that contain prolactin (PRL) mRNA or mRNA of the alpha-subunit of the glycoprotein hormones (alpha-GSU) but not the number of 'multi-hormone mRNA cells' not containing PRL or alpha-GSU mRNA. 'Multi-hormone mRNA cells' may function as 'reserve cells' in which a particular hormone mRNA may be translated under a particular physiological condition demanding a rapid increase of that hormone. PMID- 11935396 TI - Biosynthesis and secretion of pituitary hormones: dynamics and regulation. AB - Production and secretion of hormones by the pituitary involve highly orchestrated intracellular transport and sorting steps. Hormone precursors are routed through a series of compartments before being packaged in secretory granules. These highly dynamic carriers play crucial roles in both prohormone processing and peptide exocytosis. We have employed the ACTH-secreting AtT-20 cell line to study the membrane sorting events that confer functionality (prohormone activation and regulated exocytosis) to these secretory carriers. The unique ability of granules to promote prohormone processing is attributed to their acidic interior. Using a novel avidin-targeted fluorescence ratio imaging technique, we have found that the trans-Golgi of live AtT-20 cells maintains a mildly acidic (approximately pH 6.2) interior. Budding of secretory granules causes the lumen to acidify to 64 years) patients with schizophrenia (n=239) who varied in their lifetime outcome from chronic institutional care to stable community residence were tested with a neuropsychological battery that examined language and memory performance. Using a large sample of normal controls previously tested with this same assessment, normative standards for performance were developed. Schizophrenic patients were divided into cortical versus subcortical versus unimpaired groups on the basis of language and memory performance. Classification overlap ranged from a high of 30% (cortical profile) to a low of 14% (unimpaired profile). Furthermore, patients classified on the basis of verbal and memory ability areas did not differ in their performance in measures from the other ability areas. Data from a 1-year follow-up of part of the sample (n=147) suggested poor stability of the subcortical classification in particular. These data suggest that cortical versus subcortical profiles of cognitive performance in schizophrenia are inconsistent across ability areas, and are not likely to be the result of stable structural or functional brain deficits. PMID- 11935425 TI - Release from proactive inhibition in schizophrenia and its potential as a genotypic marker. AB - Release from proactive inhibition (RPI) in first-episode (FE) schizophrenic patients and the potential of RPI as a genotypic marker of schizophrenia was investigated in two studies. The first study showed that FE patients ( n=35) exhibited weaker RPI than matched obsessive compulsive disorder (OCD) patients (n=20) as well as healthy controls (n=34). OCD patients and controls showed similar RPI. The second study showed that RPI is similar in both parents of patients (n=64) and matched controls (n=52). Results were explained in terms of additional evidence for impairment in the inhibition of irrelevant information in schizophrenia. Combination of our data and literature review suggests that diminished RPI can be attributed to a combination of dorsolateral-prefrontal and dorsomedial-thalamic impairment. RPI may not be a genotypic marker for schizophrenia but rather related to the illness or its treatment. An alternative explanation in terms of insufficient power of the parent-sample, due to unilineal inheritance of schizophrenia is, however, possible. PMID- 11935426 TI - Right-left discrimination in male and female, young and old subjects. AB - The present study investigated right-left discrimination, with a paper-and-pen test using line figures. The test consists of line drawings of a person with no, one, or both arms crossing the vertical body axis of the figure. The subjects' task is to mark with a pencil, as fast as possible, which is the right or left hand in the figure. The line drawings are either viewed from the back, from the front, or randomly alternating between the back and front views. The sample consisted of 322 male and female subjects, split into four different groups, from children to old adults. The results showed increasing performance from children to young adults, with a decline in performance in the old adults (>50 years). The condition with alternating front-back views was the most difficult, particularly when the figure also had both arms crossing the vertical body midline. There were no gender differences except for better male performance in the young adults group (18-22 years). The results are discussed in relation to theories of mental rotation and lateralization of information processing strategies, in addition to right-left discrimination across the age groups. PMID- 11935427 TI - Learning, memory, and executive control in individuals with obstructive sleep apnea syndrome. AB - A range of neuropsychological deficits have been identified in individuals with obstructive sleep apnea syndrome (OSAS) and have been related to disruptions in function of the frontal cortex of the brain. We hypothesized that impairments in the use of strategic, frontally-mediated processes that facilitate learning and memory would be associated with deficits in the long-term episodic memory of verbal material (i.e., word lists). We evaluated 28 adults with OSAS and 24 controls (ranging from 28 to 60 years of age) using the California Verbal Learning Test. General executive abilities were assessed using the Wisconsin Card Sorting Test, Letter fluency, and Category fluency. Individuals with OSAS exhibited poorer recall across learning trials, less efficient use of semantic clustering, and poorer use of semantic cues. Retention of previously encoded information and recognition, however, were intact. With the exception of letter fluency, deficits were not observed in general executive control. Results are discussed within the context of disruptions in the interactions between long-term memory and executive abilities that are subserved by frontal and distal brain regions. PMID- 11935428 TI - Mental rotation in unipolar major depression. AB - Mental rotation (MR) performance may be used as an index of mental slowing or bradyphrenia, and may reflect, in particular, speed of motor preparation. MR was employed with a sample of both melancholic (n=8) and non-melancholic (n=9) unipolar depressed patients and healthy controls (n=10) to determine if motor slowing associated with depression might be reflected in slowed motor preparation (as reflected in slope of the MR function) independent of actual motor slowing (overall response time). Both melancholic and non-melancholic patients showed a generalised slowing relative to controls, perhaps reflecting bradykinesia and akinesia. This effect was significantly greater in the melancholic group than in the non-melancholic group. Relative to both the controls and the non-melancholic groups, the melancholic patients showed a progressive slowing with increasing angle of orientation indicating a specific slowing of MR. This deficit suggests a role of slowed motor planning in the psychomotor retardation of patients with melancholic depression. PMID- 11935429 TI - Sex differences in episodic memory: the influence of intelligence. AB - The influence of estimated intelligence (group assessment of WAIS-R( S)) on sex differences in face recognition, as well as verbal and non-verbal episodic memory tasks was examined in 99 women and 88 men between 20 and 40 years of age. Results showed that men performed at a higher level than women on the WAIS-R(S) subtest Information, whereas the opposite was true for Digit symbol. Women performed at a higher level than men on the verbal episodic memory tasks and on face recognition, but there were no sex differences on the non-verbal episodic memory task. Estimated intelligence for both men and women was positively associated with most episodic memory measures, the exception being face recognition in women. In face recognition, there was no association to estimated intelligence, suggesting that face recognition performance in women is unrelated to several basic cognitive processes. PMID- 11935430 TI - Selective impairments in spatial memory after ischaemic stroke. AB - There is evidence that object-location memory consists of three separate processes, that is, positional memory, binding of objects to locations, and a possible integration mechanism. A group of 26 patients with lesions following ischaemic stroke was studied to find evidence for selective impairments in these three processes using displays with pictures of everyday objects on different locations. Moreover, object-recognition memory and visuospatial construction was assessed. The Corsi Block-Tapping Task and Oxford Stylus Maze Test were included as standard spatial-memory tests. The results showed that of these 26 patients, 8 patients had selective impairments in the aforementioned object-location memory task conditions (both after immediate and delayed testing). These data further support the existence of separate cognitive processes within memory for object locations that can be selectively affected by cerebral dysfunction. PMID- 11935432 TI - Short-term chlorambucil for refractory uveitis in Behcet's disease. AB - AIM: To evaluate the effects of short-term chlorambucil therapy in the management of uveitis due to Behcet's disease. METHODS: Forty-four patients with refractory uveitis associated with Behcet's disease who had had short-term chlorambucil therapy for about 23 weeks were included in the study. The frequency of attacks (number of attacks per year) and the longest period between the attacks were analyzed to evaluate the efficiency of the therapy. The therapy was judged to be effective if the patient had < or =1 attack a year and/or > or =1 year between the attacks. RESULTS: The mean follow-up time was 51.4 +/- 32.5 months (range: 13 122 months). Following the therapy, the mean frequency of attacks had decreased from 4.9 +/- 2.3 to 0.9 +/- 1.4 (p < 0.0001) and the mean longest period between the attacks was prolonged from 4.4 +/- 2.3 months to 25.7 +/- 23.1 months (p < 0.0001). The ratio of severe attacks had decreased from 74.1 +/- 34% to 51.3 +/- 36.6% (p = 0.0218). The best-corrected visual acuity was increased in 32.9%, decreased in 34.2%, and the same in 32.9% of the eyes. Phytsis bulbi developed in three (3.8%) eyes. New attacks were seen in 56.8% of patients and another immunosuppressive agent(s) was given to 40.9% of the patients 1-8 months after treatment. No serious side effects were observed during the chlorambucil therapy. CONCLUSION: Short-term chlorambucil therapy for refractory uveitis in Behcet's disease is effective in controlling the disease in two-thirds of patients. PMID- 11935433 TI - Autoantibodies specificity in acute anterior uveitis according to the presence of the HLA-B27 allele. AB - PURPOSE: To study HLA-B27(+) and HLA-B27(-) patients with AAU with respect to the frequency and specificity of organ-specific and non-organ-specific autoantibodies. METHODS: Fifty-seven consecutive patients with AAU were subjected to ophthalmologic and rheumatologic evaluation as well as to autoantibody determination: antinuclear antibodies (ANA), rheumatoid factor (RF), anticardiolipin, anti-smooth muscle, and anti-parietal cell antibodies, and immunoblot for antibodies to HeLa cells and to bovine iris extract. HLA-B27 was determined by a microlymphocytotoxicity assay. Statistical analysis employed chi square test, Fisher's exact test, and McNemar test. RESULTS: Thirty-four patients had the HLA-B27 allele (B27(+)/AAU) and 23 did not (B27(-)/AAU). ANA, RF, anticardiolipin, and anti-parietal cell antibodies appeared in low frequency. No patient presented anti-smooth muscle antibody. Immunoblot showed a high frequency of antibodies to HeLa cell proteins in B27(-)/AAU patients with predominant reactivity at 46 kDa and 56 kDa. In contrast, sera from B27(+)/AAU patients reacted poorly against HeLa cell antigens. Immunoblot with bovine iris extract showed a significant frequency of antibodies in both groups, with a predominant response to antigens with an estimated mobility of 35, 52, and 54 kDa. CONCLUSION: Antibodies specific to iris antigens were equally frequent in both acute uveitis groups, whereas non-organ-specific autoantibodies, especially those to HeLa cell proteins, were far less frequent in B27(+)/AAU than in B27(-)/AAU patients. PMID- 11935434 TI - Uveitis in juvenile arthritis. AB - PURPOSE: To evaluate the clinical features of and determine the risk factors for uveitis in patients with juvenile arthritis. METHODS: The prevalence and clinical characteristics of uveitis were studied retrospectively in 90 children diagnosed with arthritis. Patients with uveitis were compared with those who did not have eye involvement. RESULTS: Uveitis was diagnosed in 11 patients (12.2%). Of these, seven (63.6%) had oligoarticular, two (36.4%) had polyarticular, and one (9.1%) had systemic-onset juvenile rheumatoid arthritis (JRA). One patient (9.1%) was diagnosed with enthesitis-related arthritis (ERA) (9.1%). The prevalence of uveitis was significantly higher in patients with oligoarticular JRA. The mean age at onset of arthritis in the uveitis patients was 4.39 years, which was significantly lower than in the non-uveitis group. There was no gender difference in the risk of developing uveitis. Antinuclear antibodies (ANA) was positive in seven (63.6%) of the 11 uveitis patients, confirming ANA as a significant determinant for uveitis in juvenile arthritis. Rheumatoid factor was not found to be a risk factor. One (9.1%) of the 11 patients developed serious sight threatening complications during the follow-up period. CONCLUSION: This study confirmed that oligoarticular onset, ANA positivity, and young age are risk factors for developing uveitis in patients with juvenile arthritis. Gender was not found to be a determining factor. Prompt treatment of uveitis effectively decreased the prevalence of visual impairment. PMID- 11935435 TI - Respiratory syncytial virus and chlamydia are not detectable by PCR in ongoing vernal keratoconjunctivitis. AB - Respiratory syncytial virus (RSV) and chlamydial infection may be pathogenetic factors in allergic diseases, perhaps also in ocular allergy. We analyzed the presence of RSV and chlamydial nucleic acids using reverse transcription (RT)-PCR and PCR, respectively, in conjunctival biopsies from patients with vernal keratoconjunctivitis (VKC) in order to determine whether these agents play a role in the maintenance of the disease. All biopsy samples were negative for both RSV (n = 15 for VKC and n = 10 for control subjects) and chlamydia (n = 8 for VKC and n = 7 for control subjects) homologous sequences. A direct association between RSV or chlamydial infection and ongoing inflammation in VKC could, therefore, not be confirmed. PMID- 11935436 TI - Minor influence of the immunosuppressive cytokines IL-10 and TGF-beta on the proliferation and apoptosis of human retinal pigment epithelial (RPE) cells in vitro. AB - Undesirable immune reactions such as uveitis or graft rejection after corneal and subretinal transplantations are serious inflammations in the eye. Minimizing this process by means of physiological suppressors, either through systemic or intraocular administration with or without gene therapy, is a future therapeutic possibility. In our study, we used different concentrations of transforming growth factor-beta (TGF-beta; 5, 10, and 50 ng/ml) and interleukin-10 (IL-10; 100, 200, and 500 U/ml), both known as modulators of the suppression process, to treat human retinal pigment epithelium (RPE) cells in vitro. The influence of both cytokines on the viability and proliferation of the RPE cells was measured. Furthermore, the secretion of typical markers of the apoptosis process, such as Fas, soluble Fas ligand, and bcl-2, was investigated. Our results show that the concentrations of TGF-beta and IL-10 used have only a slight influence on RPE cells. Cell proliferation under the influence of TGF-beta was significantly reduced, whereas more Fas protein could be found in the cell lysate of the IL-10 samples. In general, IL-10 seemed to have less effect on the physiology of RPE cells. The discussion of the therapeutic use of an immunosuppressive factor in the eye should therefore be focused more on this cytokine. PMID- 11935438 TI - Macular infarction in thrombotic thrombocytopenic purpura. AB - PURPOSE: To report a patient with transient acute generalized thrombotic thrombocytopenic purpura (TTP) who suffered severe permanent visual loss due to retinal vascular occlusions in both eyes. METHODS: Case report. RESULTS: Severe permanent visual loss occurred in a patient with acute transient TTP. Extensive soft exudates were seen at the fundus of both eyes. Fluorescein angiography demonstrated multiple retinal vascular occlusions associated with capillary nonperfusion. CONCLUSIONS: Severe and permanent visual loss due to retinal vascular occlusions demonstrated by fluorescein angiography occurred in a patient with transient thrombotic thrombocytopenic purpura. PMID- 11935437 TI - Uveitis with autoimmune hepatic disorders. AB - Many cases of uveitis are thought to be autoimmune in nature. We report two cases of panuveitis, one of which was associated with chronic active hepatitis and the other with primary biliary cirrhosis. This suggests that the autoimmune disturbance may have been responsible for the development of the intraocular inflammation. PMID- 11935444 TI - Policy statement on the presence of third party observers in neuropsychological assessments. PMID- 11935446 TI - Familial intelligence as an estimate of expected ability in children. AB - Estimation of premorbid or expected IQ in children from IQs of family members was evaluated for precision, sensitivity, and diagnostic efficiency. Prediction of a child's IQ from familial ability has limited precision, and the sizable discrepancies from familial IQ estimates required for statistical significance reduce their sensitivity. In a Monte Carlo simulation, familial IQ discrepancies had stronger positive than negative predictive power for distinguishing severely head-injured from normal children, with acceptable predictive power being obtained over a limited range of circumstances. Thus, while a significant discrepancy may be meaningful, the absence of one is often inconclusive. Familial IQ discrepancies should be used very cautiously, if at all, and are not substantially more accurate than discrepancies from demographic IQ estimates. PMID- 11935445 TI - Identification of malingered head injury on the wechsler adult intelligence scale - 3rd edition. AB - Head injured patients show an IQ subtest pattern that can be discriminated from the profile produced by individuals who attempt to malinger intellectual decline due to head trauma. The current paper demonstrates that previously replicated methods for making this discrimination on the WAIS - R generalize to the WAIS - 3. The discriminant function equation accurately classified 83% of nonlitigating head-trauma patients with documented injuries and 72% of persons simulating intellectual impairment due to head trauma. A total of 45% of litigating mild head-trauma patients with purported intellectual decline but no documented loss of consciousness, hospitalization, or CT abnormality were classified as malingering by the discriminant function. A Vocabulary-Digit Span difference score provided 71% overall diagnostic accuracy, and may be informative when screening profiles by visual inspection or when complete WAIS - 3 results are unavailable. PMID- 11935447 TI - Classification accuracy of the portland digit recognition test in traumatic brain injury. AB - This study examined the classification accuracy of the Portland Digit Recognition Test (PDRT) in traumatic brain injury (TBI). It differs from past studies in assigning patients to malingering and control groups on the basis of compensation seeking status and the presence of external markers for malingering. Sensitivity and Specificity were.77 and 1.00, respectively. Past research comparing compensation-seekers to noncompensation-seekers reported Sensitivities of.33 or lower (Specificity is always high). This study demonstrates that past research has seriously underestimated the Sensitivity of the PDRT and raises questions about the true Sensitivity of other malingering techniques as well. PMID- 11935448 TI - Practice effects on the Wisconsin Card Sorting Test-64 Card version across 12 months. AB - The Wisconsin Card Sorting Test (WCST: Heaton, Chelune, Talley, Kay, & Curtiss, 1993) is among the most commonly administered measures of executive function. Recently, a short form of the test was developed (WCST-64: Kongs, Thompson, Iverson, & Heaton, 2000), and it affords psychometric properties commensurate with the full version of the test. Yet, similar to other measures of executive function, relatively little is known concerning the effects of repeated administration on the WCST-64. Towards this end, 53 men (age M = 32.38) were administered the WCST-64 twice over 12 months, and scores on several indices improved significantly during this interval. Suggestions concerning the use of these measures in longitudinal research designs and clinical follow-up examinations are offered, and reliable change indices concerning these measures are included. PMID- 11935449 TI - Executive functioning in adult attention-deficit hyperactivity disorder. AB - The present study examined the executive abilities of 35 adults diagnosed with Attention-Deficit Hyperactivily Disorder (ADHD) and 32 adults without the disorder (n = 67) who were equivalent in age, gender, years of education, and Full Scale IQ. The ADHD group performed significantly worse on Stroop Color-Word (eta(2) =.18) and Interference (eta( 2) =.08), as well as time to complete Trails B (eta(2) =.08) than the controls (all ps <. 05). Analysis of Design Fluency indicated that the ADHD group committed more perseverative (eta(2) =.06) and non perseverative (eta(2) =.12) errors than did controls; however, novel output was equivalent for the groups. No group differences were observed on tests measuring cognitive initiation, abstract thinking, or working memory (all ps >.30; eta(2) =.00-.01). The distributions of WCST variables showed severe skew associated with high-functioning performance on the test among both groups. The pattern of results suggests the presence of specific deficits in response inhibition, with intact abilities in other cognitive domains, such as primary verbal and visuospatial skills. These findings are consistent with the literature on neuropsychological deficits among children with ADHD. That persons with ADHD present a primary deficit of behavioral inhibition supports Barkley's (1997) theory of ADHD, as opposed to theories by Denckla (1996) and Roberts and Pennington (1996) that emphasize intention and working memory. PMID- 11935450 TI - Validation of the computerized assessment of response bias in litigating patients with head injuries. AB - The detection of malingering in neuropsychological assessment requires valid measures of incomplete effort. The Computerized Assessment of Response Bias (CARB) is a digit-recognition procedure designed to detect poor effort during a neuropsychological evaluation. We examined the CARB performance of a consecutive series of litigating patients (N = 119) with a full range of head-injury severity, from trivial to severe. Patients with trivial or mild head injuries performed more poorly than did patients with frank brain injuries. Specifically, patients with moderate to severe traumatic brain injuries obtained higher total scores and demonstrated briefer response latencies than did patients with trivial or mild head injuries. PMID- 11935451 TI - Construct validity of selected Automated Neuropsychological Assessment Metrics (ANAM) battery measures. AB - The Automated Neuropsychological Assessment Metrics (ANAM) is a computerized library of tests designed to assess neurocognitive functioning across administrations (Kane & Reeves, 1997). This study was designed to examine neuropsychological constructs measured by selected ANAM measures and to compare them with traditional measures putatively assessing similar domains. The sample consisted of 191 outpatients with suspected neurocognitive dysfunction. Correlations and regressions indicated significant relationships between traditional and computerized tests measuring similar constructs. PCA results yielded a three-factor solution: Factor I (Processing Speed/Efficiency), Factor II (Retention/Memory), and Factor III (Working Memory). PMID- 11935452 TI - Preserved semantic access in global amnesia and hippocampal damage. AB - C.B., a right-handed 33-year-old man, presented with anterograde amnesia after acute heart block. Cognitive abilities were normal except for serious impairment of long-term episodic memory. The access to semantic information was fully preserved. Magnetic resonance showed high signal intensity and marked volume loss in the hippocampus bilaterally; the left and right parahippocampal gyrus, lateral occipito-temporal gyrus, inferior temporal gyrus, and lateral temporal cortex were normal. This case underlines that global amnesia associated with hippocampal damage does not affect semantic memory. Although the hippocampus is important in retrieving context-linked information, its role is not so crucial in retrieving semantic contents. Cortical areas surrounding the hippocampus and lateral temporal areas might guide the recall of semantic information. PMID- 11935453 TI - Performance characteristics of postacute traumatic brain injury patients on the WAIS-III and WMS-III. AB - Publication of the third editions of the Wechsler intelligence and memory batteries in 1997 created a need for research identifying Wechsler Adult Intelligence Scale-Third Edition/Wechsler Memory Scale-Third Edition (WAIS III/WMS-III) profile patterns associated with neuropathology. The WAIS-III/WMS III Technical Manual offers data on various diagnostic groups, including traumatic brain injury (TBI). Hawkins (1998) employed Technical Manual data to propose certain diagnostic guidelines. In order to validate the conclusions put forth by Hawkins as they apply to brain injury, we examined WAIS-III and WMS-III profiles in an independent sample of 46 TBI cases. As expected, the WAIS-III Processing Speed Index (PSI) was more sensitive to brain injury than other WAIS III composites; and specific WAIS-III scores were stronger than certain WMS-III scores. On the other hand, the predicted relationship for WMS-III auditory and visual indexes was not found. The lack of specificity for TBI of the proposed index comparisons confirms the need to validate such hypotheses in independent samples. PMID- 11935454 TI - Letter cancellation performance across the adult life span. AB - Letter cancellation tasks are paper and pencil tests widely used in clinical and research settings as quick measures of attention/concentration, visual-spatial scanning abilities, and visual-spatial dysfunctions such as spatial neglect. Despite their popularity, only a few studies have investigated effects of age on letter cancellation performance and no comprehensive large-scale adult life-span norms are available for any of the letter cancellation tasks. We have developed a letter cancellation test, and we report a normative data obtained from a partially stratified sample of 351 healthy adults between 18 and 91 years of age. The results show the expected large age-related decline in the speed of letter cancellation performance and no age-related differences in spatial distribution of cancellation errors. We also provide various equations for precise predictions of Cancel H test performance. The results of correlational analyses show high reliability and provide evidence for both convergent and divergent validity of our letter cancellation task. PMID- 11935455 TI - Lexical emotional expression across the life span: quantitative and qualitative analyses of word list generation tasks. AB - The current study examined the effects of age and gender on emotional and nonemotional expression using an experimental word list generation (WLG) task (also referred to in the literature as verbal fluency) from the New York Emotion Battery (Borod, Welkowitz, & Obler, 1992). Subjects were 28 young ( M = 29.6 years), 28 middle-aged (M = 49.8 years), and 28 older (M = 69.9 years) healthy adults. The WLG task consists of 8 emotional (E; 3 positive and 5 negative) and 8 nonemotional (NE) categories. We developed and present here a detailed word error type analysis that was used to evaluate the lexical output. In this study, both quantitative (amount of output and error-types) and qualitative (accuracy and intensity) analyses were used. While subjects produced more nonemotional than emotional words and more positive than negative words, the amount of error-free output and the number of errors did not change with age. An age group by error type interaction indicated that older adults, especially men, produced more repetition errors than younger adults. The error-free output was subsequently rated for accuracy and emotional intensity. The rating data revealed that older women's overall lexical output was less accurate than that produced by younger women. Also, negative emotional words were more accurate and intense than positive emotional words. The procedures described here have implications for research assessing word list generation and emotional expression in clinical populations. PMID- 11935456 TI - A computer-assisted administration of the Continuous Visual Memory Test. AB - The Continuous Visual Memory Test (CVMT) is a continuous design recognition test designed for the assessment of visual memory for clinical and research applications. Comparisons were made between the traditional and a computer assisted format of the CVMT using 51 university students. Formats were compared on five CVMT dependent measures, concurrent validity, split-half reliability, and selected dependent measures from the California Verbal Learning Test (CVLT). Participants scored higher on the computer format dependent measures, with the exception of false alarms (new items incorrectly identified as old), which remained comparable between formats. Adequate concurrent validity (r =.70) and split-half reliability (rs =.61-.70) were demonstrated for CVMT Total Score. Restriction of range and ceiling effects precluded reliability and validity conclusions for CVMT Delay. The computer format showed a nonsignificant trend towards lower correlations with CVLT dependent measures. PMID- 11935459 TI - Insights into the role of immune activation in HIV neuropathogenesis. AB - How does HIV infection lead to the development of central nervous system disease? Central to this question is identification of the relative contributions of (1) the virus, (2) its host cells, and (3) secondary or downstream events to the pathological process. These are re-examined in this brief review. Also, a greater appreciation for the role of systemic events in neuroinflammation is emerging, with likely relevance to HIV-associated dementia. We propose here a model for HIV neuropathogenesis that highlights the role of systemic monocyte activation and subsequent neuroinvasion in initiating the disease. PMID- 11935460 TI - Murine hepatitis virus--a model for virus-induced CNS demyelination. AB - Most murine hepatitis virus (MHV) strains, as their name suggests, infect the liver. However, several murine strains are tropic for the central nervous system (CNS) and cause encephalitis with subsequent CNS demyelination. The CNS demyelination shares pathological similarities with human CNS demyelinating diseases such as multiple sclerosis (MS). These viruses are, therefore, used to study the role of the immune system in viral clearance from the CNS, in CNS demyelination, and in remyelination. Nevertheless, it is still unclear exactly how MHV induces demyelination and to what extent the immune system plays a role in this pathology. Here we review this field in the context of the immune response to MHV in the liver and the CNS focusing on studies that have been published in the past 5 years. PMID- 11935461 TI - Lentiviral expression of HIV-1 Vpr induces apoptosis in human neurons. AB - Our recent studies have demonstrated that extracellular, recombinant human immunodeficiency virus type I (HIV-1) Vpr protein is highly neurotoxic in the microenvironment of differentiated mature human neurons and undifferentiated neuronal precursors. Although most of the direct neurotoxic effects of HIV-1 have been attributed previously to the envelope gene product, gp120, and the Tat regulatory protein, it was demonstrated that Vpr protein caused apoptosis comparable to that induced by gp120 protein in a dose-dependent manner in the neuronal system. Having observed the neurocytopathic effects of extracellular Vpr protein previously, the effects of virally expressed Vpr on nondividing, terminally differentiated human neurons were investigated. An HIV-1-based three plasmid expression vector system was utilized to study the effects of intracellularly expressed Vpr. These virion preparations were then used to transduce neurons generated from the human neuronal precursor NT2 cell-line. Intracellularly expressed Vpr induced apoptosis within terminally differentiated neurons, as demonstrated by TUNEL assays. Additionally, virions lacking Vpr expression did not significantly induce apoptosis within these neurons. These results suggest that HIV-1 Vpr may also be leading directly to selective neurotoxicity through intracellular expression. Furthermore, human apoptosis gene microarray comparisons exhibited an up-regulation of Bcl-2-related mRNA, as well as other apoptosis genes involved in the mitochondrial apoptotic pathway, for the Vpr-transduced neuronal cells, when compared to Vpr-negative controls. Thus, Vpr delivered intracellularly, as well as extracellularly, is involved in the induction of significant neuronal apoptosis and may be one of the molecular mechanisms in HIV-1-induced encephalopathy. PMID- 11935462 TI - Mutation in loop I of VP1 of Theiler's virus delays viral RNA release into cells and enhances antibody-mediated neutralization: a mechanism for the failure of persistence by the mutant virus. AB - The DA strain of Theiler's murine encephalomyelitis viruses (TMEV) causes a central nervous system (CNS) demyelinating disease with viral persistence despite the presence of high serum anti-TMEV antibody titers. The DA virus mutant, T81D, was created to have a mutation at position 81 in loop I of VP1, close to the putative virus receptor binding site. T81D showed slower replication in vitro and in vivo. T81D-infected mice developed anti-TMEV antibody responses with no virus persistence. We tested whether the differences between the viruses were due to alteration in virus-cell interactions, or in the resistance to neutralization by anti-TMEV antibody. Using radiolabeled viruses, we found no difference in binding to permissive cell lines between the mutant and wild-type viruses. In a semipermissive cell line, DA virus bound more efficiently than T81D. During the uncoating step, both viruses decapsidated without the production of stable intermediates and 80% of viruses were eluted or decapsidated after 45 minutes. At the final step of uncoating, however, T81D showed a slower rate of RNA release than DA virus into cells using a photoinactivation assay. Anti-TMEV monoclonal and polyclonal antibodies neutralized T81D virus more efficiently than DA virus in suspension. Further, these anti-TMEV antibodies were able to neutralize viruses that had already attached to cells but not internalized (postadsorption neutralization [PAN]). However, DA virus showed significant resistance to PAN after cells were incubated at 37 degrees C compared with T81D-infected cells. The development of resistance to PAN appeared to correlate with the rate of RNA release from virions into cells. In T81D virus infection, the slow RNA release and high susceptibility to neutralization by antibodies would result in a failure to establish virus persistence in vivo. Conversely, rapid RNA release and resistance to neutralization could favor virus persistence in DA virus infection. PMID- 11935463 TI - Isolation and growth of a cytopathic agent from multiple sclerosis brain tissue. AB - Although many studies support a role for viruses in multiple sclerosis (MS) etiopathology, no specific agent has been consistently associated with significant numbers of MS patients without concomitant detection in non-MS controls. Previous studies have shown the presence of viral-like structures in MS plaques, although the specificity of these structures for MS has been questioned. The present study describes the use of polyclonal antisera against feline and human brain-derived cytopathic agents and immunoaffinity chromatography to purify and partially characterize possible virus-like structures from MS brain tissue. Chromatography eluates from 4 MS brains contained pleomorphic particles up to 350 nm in diameter and tubular/filamentous-like structures approximately 10-18 nm in thickness. Inoculation of primary rat glial cell cultures with chromatography eluates from MS brain tissue resulted in a reproducible pattern of cytopathic effects in the form of multinucleation in cells identified immunocytochemically as oligodendrocytes. Antisera raised against the feline and MS-derived cytopathic agents were used to successfully immunolabel infected oligodendrocyte-like cells and syncytia and to detect a 66,000 M(r) protein on Western blots of inoculated cultures or concentrated MS brain eluates. Similar structures, cytopathic effects (CPE) and protein expression were not observed in eluates from 5 control brains or in cultures inoculated with control brain eluates. These studies demonstrate that cytopathic, virus-like structures can be isolated from MS brain tissue using antisera raised against a cytopathic agent rescued from demyelinating brain lesions in cats. The identity of this agent and its possible role in MS aetio pathology remains unknown. PMID- 11935464 TI - Rapid spread of a neurovirulent strain of HSV-1 through the CNS of BALB/c mice following anterior chamber inoculation. AB - Following uniocular anterior chamber (AC) inoculation of BALB/c mice with the KOS strain of herpes simplex virus type 1 (HSV-1), virus spreads from the injected eye to the ipsilateral suprachiasmatic nucleus (SCN) in the central nervous system (CNS) to infect the optic nerve and retina of the contralateral eye, and mice develop retinitis in that eye only. In contrast, after AC inoculation of BALB/c mice with the H129 strain of HSV-1, mice develop bilateral retinitis. The pathway(s) by which H129 spreads to cause bilateral retinitis is not known. To determine the route and timing of H129 spread after AC inoculation, BALB/c mice were injected in the AC of the right eye with 5 x 10(3) PFU of H129. Brains from 30 mice were sectioned on a brain matrix and the amount of virus in the brain and eyes was determined by plaque assay. Frozen sections were prepared from the eyes, brain, and trigeminal ganglia of an additional 30 mice, and HSV-1 antigen was detected by immunohistochemistry. After AC inoculation, H129 follows a pathway similar to KOS in the CNS, but H129 appears to spread more rapidly than KOS within the CNS. Unlike KOS, H129 is able to infect brain stem nuclei and H129 infected mice developed neurological impairments in addition to bilateral retinitis. The results of these studies suggest that the ability of H129 to spread rapidly in the CNS allows early virus infection of retino-recipient nuclei proximal to the contralateral and ipsilateral optic nerves. Early infection of retino-recipient nuclei, such as the SCN may allow virus to spread into the retinas before a virus-specific immune response can be induced. PMID- 11935466 TI - Further in vitro characterization of mouse hepatitis virus papain-like proteinase 1: cleavage sequence requirements within pp1a. AB - Proteolytic processing of the mouse hepatitis virus strain A59 (MHV-A59) replicase gene product, pp1a, results in polypeptides p28, p65, p50, and p240 in infected cells. Based on previously identified p28 and p65 cleavage sites, a p50 cleavage site was proposed to occur between Ala-1262 and Ala-1263. Results of mutagenesis and in vitro cleavage assays show that PLP-1 was able to cleave in trans when the proposed p50 cleavage sequence replaced the p28 cleavage sequence. Mutagenesis was also used to investigate cleavage between Gly-904 and Val-905, a cleavage site predicted to produce a precursor of p65, p72, that was detected in cells infected with MHV strain JHM, but not with MHV-A59. No cleavage could be detected using substrate that carried both the p65 site and the predicted p72 cleavage sequence. Thus, it appeared that PLP-1 could recognize the proposed p50 sequence but not the predicted p72 site under the in vitro conditions used. PMID- 11935465 TI - HIV-associated cognitive impairment before and after the advent of combination therapy. AB - The objective of this study was to describe the occurrence of HIV dementia and neuropsychological testing abnormalities in a new cohort of HIV-seropositive individuals at high risk for HIV dementia and to compare these results to a cohort before the advent of highly active antiretroviral therapy (HAART). HAART has been associated with improvements in cognitive performance in some HIV infected patients. However, it is uncertain whether HAART has changed the frequency of specific neurocognitive abnormalities. Baseline data from 272 HIV seropositive subjects in the Dana cohort recruited from January, 1994, to December, 1995, and 251 HIV-seropositive subjects in the Northeastern AIDS Dementia Consortium (NEAD) cohort recruited from April, 1998, to August, 1999, were compared. Participants in both cohorts received nearly identical assessments. After adjusting for differences in age, education, gender, race, and CD4 count between the two cohorts, there were no differences in the occurrence of HIV dementia or abnormalities either 1 SD or 2 SDs below established norms for any of the neuropsychological tests. Even though HAART has reduced the incidence of HIV dementia, HIV-associated cognitive impairment continues to be a major clinical problem among individuals with advanced infection. PMID- 11935467 TI - Differential use of the nicotinic receptor by rabies virus based upon substrate origin. AB - To determine the role that the neuronal nicotinic acetylcholine receptor plays in the adsorption process of rabies virus (RV), adult dorsal root ganglion dissociated cultures were exposed to nicotinic agonists before being inoculated. The fixed strain of RV Challenge Virus Standard-11 (CVS-11) was used after being passaged in two different ways, in baby hamster kidney (BHK) cells and in adult mouse brain (MB). Carbachol and nicotine reduced the percentage of CVS-MB infected neurons, yet none of the agonists tested changed the proportion of CVS BHK infected neurons. This result suggests that the RV phenotype changes depending on its replication environment and neuronal nicotinic acetylcholine receptors are preferentially used for infection by RV strains adapted to adult mouse brain but not to fibroblasts. PMID- 11935469 TI - [Homocysteine, methylenetetrahydrofolate reductase/C677T genotype and risk for coronary heart disease. The AtheroGene study]. PMID- 11935470 TI - [Skin nodules and ulcers of the limbs in a patient with rheumatoid arthritis]. AB - CASE HISTORY: While being treated with corticosteroids and methotrexate for rheumatoid arthritis, a 63-year-old man developed livid nodules on his lower arms, hands and feet, as well as fever, necrotizing skin ulcers and rupture of a finger extensor tendon. INVESTIGATIONS: No vasculitis was found in a biopsy of one of the nodules on the lower arm. Fast growing mycobacteria, classified as M. marinum by PCR, were cultured from wound swabs. TREATMENT AND COURSE: The lesions healed on administration of ciprofloxacin, ethambutol and clarithromycin as well as local treatment. CONCLUSION: Cutaneous lesions of an atypical mycobacterial infection are often misdiagnosed. This is especially so in immunocompromised patients and in the differential diagnosis of vasculitis. PMID- 11935472 TI - [Ventilator-associated pneumonia--prevention and diagnosis]. PMID- 11935471 TI - [Ventilator-associated pneumonia--case report]. PMID- 11935473 TI - [Ventilator-associated pneumonia--treatment]. PMID- 11935475 TI - [T-cell therapy in oncology. Therapeutic vaccination, allogeneic blood stem cell transplantation and adoptive T-cell transfer]. PMID- 11935476 TI - [Sepsis]. PMID- 11935477 TI - [Shoulder surgery - state of the art and future development]. PMID- 11935479 TI - [New treatment approaches in sepsis]. AB - The treatment of sepsis consists of focus control as well as supportive and adjuvant therapy. Especially the last option has been investigated during the last years. Different approaches showed promising results in animal experiments and phase-I trials but did not prove to be successful in large multicenter studies. The application of TNF-receptors or interleukin-1 receptor antagonists did not lead to an improvement of outcome in patients with sepsis. Most studies with TNF-antibodies also presented negative results. However, a recent large study with a monoclonal antibody against TNFalpha demonstrated a significant survival benefit. The recently published PROWESS study is the first investigation demonstrating the decrease of mortality in patients with sepsis after administration of protein C. Additionally, current data support the low-dose hydrocortisone therapy in patients with vasopressor dependent septic shock. PMID- 11935478 TI - [Diagnostic approach to sepsis - state of the art]. AB - Early diagnosis of the different severities of septic inflammation is important for early implementation of specific therapies. Sepsis and severe sepsis are accompanied by clinical and laboratory signs of systemic inflammation. However, patients suffering from non-infectious inflammation may present with similiar signs and symptoms making it difficult to diagnose infection based on clinical findings alone. Bacteriological evidence of sepsis, though definitive and specific, may not be obtainable, is time-consuming and even may not occur concurrently with clinical signs of sepsis. It is therefore important to identify markers, which, by enabling an early diagnosis of sepsis and organ dysfunction, would allow early specific therapeutic interventions. Wheras C-reactive Protein is a more sensitive parameter for the diagnosis of non-systemic infections, Procalcitonin seems to be a useful parameter to improve the diagnosis and monitoring of therapy in patients with severe sepsis and septic shock. PMID- 11935480 TI - [Arthroscopic treatment of shoulder instability]. AB - Arthroscopic treatment of glenohumeral instability has become more common over the last years. Compared with open reconstruction which still is the gold standard in the treatment of glenohumeral instability the arthroscopic techniques show several advantages but also disadvantages that must be proved for every single case. Careful patient selection with regard of prognostic preoperative and intraoperative factors is substantial for successful treatment. When indicated using clinical, radiological and arthroscopic findings the results of arthroscopic stabilization could be improved over the last years and failure rates could be decreased. PMID- 11935481 TI - [Etiology of rotator-cuff-tears and consequences for legal assessment]. AB - Traumatic or degenerative etiology of rotator-cuff-tears is a matter of controversial discussion in legal proceedings. This article presents a review on the literature about current knowledge of anatomy, biomechanics and pathogenesis of rotator-cuff pathology. According to this a catalogue of potentially adequate and inadequate trauma mechanisms is proposed. Furthermore clinical history and examination as well as sonographic and radiologic findings and intraoperative pathology may help to distinguish between traumatic and degenerative changes of rotator-cuff lesions. The keypoints for and against traumatic etiology of rotator cuff-tears are mentioned that could help the expert in the preparation of an adequate expert opinion. PMID- 11935482 TI - [Current concepts in treatment of rotator cuff tears]. AB - The expectation of a functional shoulder joint, even in advanced age, the growing experience and success in shoulder surgery have brought the operative treatment another step further. The goal of any advanced knowledge should be a proper treatment of the patients and the pathology. Complex rotator cuff tears can be repaired by means of muscle transfer and partial reconstruction. In cases of irreparable rotator cuff tear arthropathy prosthetic replacement of the gleno humeral joint can be performed. In this article a review on the different techniques and indications for rotator cuff repair is given. PMID- 11935483 TI - [AC Joint Injuries]. AB - There is still controversy about the treatment for acute type III acromioclavicular joint dislocation according to Tossy and Rockwood's classification. Good functional results were reported on following operative and non-operative treatment. According to the literature the functional outcome is similar. Following surgical repair, however, complications occur more often and the period of rehabilitation is longer. The disadvantages of non-operative treatment include a higher rate of a persisting prominence of the distal clavicle. Currently available data in the literature indicate that based on the criteria of evidence based medicine non-operative treatment seems to represent the method of choice for type III injuries of the acromioclavicular joint. PMID- 11935484 TI - [Proximal humeral head fractures--Future development of shoulderimplants]. AB - In 1951 a shoulder prosthesis was developed for the treatment of complex humeral head fractures by Neer. The Neer-II-Prosthesis, which was developed further in 1973 represents the basis of modern shoulder implants. The unsatisfactory postoperative functional results for primary humeral head replacement required the development of new adaptable implants for the treatment of complex humeral fractures, which allow the reconstruction of humeral length, the reconstruction of center of rotation and humeral offset, the anatomic positioning, the reconstruction of retrotorsion and the secure refixation of the tuberosities. The development of third and fourth generation of humeral head prosthesis achieves a three-dimensional adaptability for the conditions of posttraumatic arthritis. Primary and secondary humeral head replacement in humeral head fractures as well as revision of prosthetic replacement are complex and technical demanding operations with high and different requirements on implants having to be realised on further development. PMID- 11935485 TI - [Combination of ipsilateral supracondylar humeral- and forearm fractures in children]. AB - In an 11-year period, from 01. 08. 1987 to 31. 08. 1998, a total of 72 children (mean age 7.6 years, range 2-12 years) with dislocated supracondylar humeral fractures were treated surgically in the Department for Traumatology, University Hospital, Essen. The combination of supracondylar humeral fracture and ipsilateral forearm fracture occurred in 8 children (11.1 %). 4 revealed a complete forearm fracture in the distal third, 4 children a fracture of the distal physis (Salter-Harris type II). The supracondylar humeral fractures were reduced openly via a single lateral approach and stabilized by crossed K-wire fixation. The distal forearm fractures were treated by closed reduction and percutaneous pinning. Fractures of the distal physis were treated by closed reduction and application of an above elbow cast. Excellent results were achieved in all children with ipsilateral supracondylar and forearm fractures. PMID- 11935487 TI - [Experiences with 925 outpatient operations of carpal tunnel syndrome]. AB - In the era of cost containment in our health care system the demand is made of increasing outpatient surgery e. g. for carpal tunnel syndrome. To show the possibilities of carpal tunnel release performed under outpatient conditions we analysed 925 consecutive cases during 1. 1. 1981 to 30. 4. 2001. A tourniquet was used in all cases. In 814 cases we used axillary plexus anaesthesia. 107 surgical interventions were done under local anesthesia. Two of these patients could not tolerate the tourniquet. Analgesia was complete in all but two cases. In two cases revision due to after bleeding was necessary. There were no further complications seen. The mean time for return to work was 21.3 days (min: 14, max: 36 days). The mean follow-up for these patients was 36 months. Numbness and paresthesias were relieved in 98 %, pain was relieved in 90 %, motoric weakness was relieved in 95 % of the hands. The overall satisfaction rate was 94 %. Carpal tunnel release can be performed safely and unexpensively under outpatient conditions. PMID- 11935486 TI - [Differential therapy in radial head fractures]. AB - The therapeutic regimen of radial head fractures, especially of displaced and comminuted types is controversial. The radial head resection has been critically reviewed over the past years. From 1984-1993 and 1996-1999, 105 radial head fractures were treated in our hospital. 74 were subject to clinical and radiological follow-up. Fracture-types were classified according to Mason. Undisplaced fractures were treated conservatively, displaced 2-fragment-fractures by an open reduction and screw fixation, and multifragment-fractures by a radial head resection. The results were studied on a functional and radiological basis using the "Functional Rating Index" of Broberg and Morrey and the radiological Score of Albrecht and Ganz. After conservative therapy over 80 % achieved excellent and good as well as 12.5 % satisfactory and 6.3 % unsatisfactory results. After reduction and internal fixation again 80 % had excellent and good results. After radial head resection excellent and good results were achieved in 54.6 % of the cases, satisfactory results in 24.2 % and in 21.2 % unsatisfactory results, however prognosis-influencing concomitant injuries were often present in the latter group. Using the right indication and technique, the radial head resection still is a recommendable therapeutic procedure with an altogether good prognosis. This especially applies to isolated radial head fractures where excellent and good results can be achieved in approximately 70 %. PMID- 11935488 TI - [Primary nerve transplantation: does it make sense in defect injury of peripheral nerves?]. AB - Regeneration of damaged nerves limits the outcome of complex injuries of the upper extremities. We report the follow-up results in 7 of 10 patients over 5 years who underwent a transplantation of n. suralis in 6 cases and n. cutaneus antebrachii ulnaris in 4 cases. The average DASH-score of 30 points, no neurinoma and a protective sensibility in 5 cases are comparable with the literature. PMID- 11935489 TI - [Does hospital structure influence the outcome of operative treatment of femoral neck fractures?]. AB - Data on 32 007 patients suffering from a medial fracture of the femoral neck have been collected between 1993 and 1999 in a database for external quality assurance organized by the chamber of physicians in Westfalia-Lippe. A statistical analysis (ANOVA, chi-square-test) has been performed to find out whether factors like specialization, annual volume or level of the hospital (primary, secondary or tertiary hospital) influence the outcome. RESULTS: Patients with higher preoperative risk-factors are treated more often in primary hospitals. These clinics perform conservative treatment significantly more often than tertiary hospitals (6.5 % vs. 3.8 %). Osteosyntheses are performed more often in departments specialized in traumatology (13 %) or tertiary hospitals (16.8 %). Preoperative length of stay was 0.5-0.7 days shorter in these hospitals. There is no significant difference in postoperative complications all together (23.2-25.6 %), but a significantly lower rate in postoperative complications after osteosynthesis performed by departments specialized in traumatology (11.3 % vs. 18.8 %). A volume load of more than 50 cases per year correlates with a significant decline in postoperative complications (22.5 % vs. 28.2 %). Risk adjusted mortality does not show significant differences among the different levels of hospitals. CONCLUSIONS: There are distinct differences regarding the way of treatment and procedural quality, but not concerning the short-term outcome between hospitals of different levels. PMID- 11935490 TI - [Quality of life after foot injuries]. AB - OBJECTIVE: Assessment of quality of life after foot injuries. METHODS: All patients with foot injuries treated operatively from July 1995 to December 1999 were asked to complete the SF-36 health survey. Values of four subgroups (male with foot injuries without any other injury, multiple injured male with foot injuries, male without any other injury and both calcaneus fractures or other injuries) were compared with the standardized German population (Mann-Whitney-U Test, alpha = 0.05). RESULTS: 110 of 201 patients completed the questionnaire (follow-up 54.7 %). Values of most of the subscales showed significant differences in comparison with the standardized population for three of four groups. Men with calcaneus fractures revealed the highest disabilities especially for the pain subscale. Quality of life in multiple injured patients is dominantly influenced by foot injuries. CONCLUSIONS: Foot injuries cause a strong limitation of quality of life. The SF-36 health survey provides a very suitable outcome measurement for foot injuries. PMID- 11935491 TI - [A giant malignant tumour of the skull-Case report and differential diagnosis]. AB - The case of a 72-year-old woman with a high-partially located tumor grown within a half year to a magnitude of 8.5 x 11 x 11 cm is reported. The patient remembered a mastectomy and axillary lymphadenectomy followed by chemotherapy and radiation 8 years ago. Therefore we assumed a skeletal metastasis of a breast cancer. After wide excision, an unusual morphology was found, allowing only a classification as a pleomorphic sarcoma. Searching for the pathohistological evaluation of the former breast tumor, a cystosarcoma phylloides malignum could be found out. The tumor described here can be identified as a metastasis of this rare neoplasm. PMID- 11935492 TI - [Midwifery and gynecology--View of a German Editor]. PMID- 11935493 TI - Therapy of female sexual dysfunction. PMID- 11935494 TI - [New aspects by the therapy of ovarian cancer--What changes after the ASCO Meeting 2001]. AB - The standard primary therapy of advanced ovarian cancer consists of platinum derivatives in combination with paclitaxel. In a first report presented at this year's meeting of the American Society of Clinical Oncology (ASCO) docetaxel appeared to be as effective as paclitaxel in combination with carboplatin, while the toxicity profile present some advantages. The addition of a third drug to platinum-taxane-combination does not appear to improve the therapeutic index. According to the data of the ICON1- and ACTION-trial, carboplatin containing chemotherapies are associated with a significant benefit also in early ovarian cancer. Platinum-resistant disease remains a therapeutic challenge, since the available drugs display only limited activity of short duration. Some experimental data suggest evidence for a possible therapeutic role of small molecules that inhibit the epidermal growth factor receptor (OSI-774) or antisense oligonucleotides interfering with EGF-receptor signalling (ISIS 5132). Novel classes of chemotherapeutic agents, including the acylfulvenes and the epothilone-analogues warrant further study in this disease. Multimodal therapies combining cytotoxic agents with antibodies against CA 12-5 or metalloproteinase inhibitors have failed to demonstrate any survival benefit in patients with ovarian cancer. Despite significant progress in the last decade ovarian cancer remains the most lethal gynaecologic malignancy in women in most western countries. Further multicenter clinical studies are needed to better define new therapeutic options. PMID- 11935495 TI - Magnetic Resonance Female Breast Imaging (MRFBI) - Evaluation of the Changes in Signal Intensity over Time Pre- and Post-administration of 0.2 mmol/kg Gd-DTPA. AB - PURPOSE: We are looking for a threshold value to discriminate between benign and malign breast lesions in MRI of female breast after administration of 0.2 mmol Gadolinum-DTPA/kg bw. MATERIALS AND METHODS: Double coil breast MRI (1.5 Tesla) were performed in 65 patients with an suspicious lesion for malignancy in an anteriorly examination. 57 patients could be evaluated in our study design. Histopathological 35 patients had an invasive carcinoma, 3 patients had an in situ-carcinoma and in 27 patients benign changes were found. RESULTS: For different carcinoma diameters we found a different increase of signal intensity (SI): small carcinoma (< 10 mm) had an maximum increase of SI of 102 %, medium sized (10 to 20 mm) 222 % and carcinomas over 20 mm showed an increase of 271 %. We did not find a significant difference between SI in benign and malign lesions. The sensitivity was 94.6 % the specificity 65 %. CONCLUSION: A threshold value to distinguish between malign and benign in MRI could not be defined. With the double normal Gd-DTPA dose we do not have better specificity and sensitivity than for normal dose (0.1 mmol/kg bw) is described. PMID- 11935496 TI - [Detection of human papillomavirus DNA using hybrid capture does not allow sufficient triaging of recurrent atypical pap smear classified as Pap III D]. PMID- 11935497 TI - [Uteroplacental insufficiency in chorangiomatosis]. AB - Chorangioma is a benign angiomatous tumor of the placenta occurring with a frequency of approximately 1 % of all examined placentas. Pregnancies especially with large chorangiomas are frequently complicated by abruptio placentae, premature labour, fetal anaemia, hydrops, cardiomegaly and polyhydramnios. We here report a case of an extremely rare chorangiomatosis. Multiple chorangiomas measuring up to 1.5 cm comprised approximately one third of the placenta. A caesarean section had to be performed at 36 weeks of gestation because of uteroplacental insufficiency with oligohydramnios. Displacement of normal placental parenchyma by chorangiomatosis as a cause of uteroplacental insufficiency is very likely. Colour doppler imaging allows early diagnosis and management of chorangiomas. Thus, in our case early diagnosis of uteroplacental insufficiency was rendered. Principally, early diagnosis of fetal hazard due to myocardial insufficiency of the fetus is feasible. In-utero-endoscopy permits prenatal laser devascularisation of large chorangiomas. In the reported case the pregnancy was closely monitored with colour flow imaging. The diagnosis of uteroplacental insufficiency was established because of abnormal blood flow of the uterine arteries and abnormal fetal heart rate (FHR). At the same time fetal cardiomegaly and pathologic blood flow of the ductus venosus in terms of beginning fetal myocardial insufficiency was recognized. Consequently, caesarean section could be performed opportune. PMID- 11935499 TI - Immunological and biological activity of different commercial preparations of human chorionic gonadotropin. AB - In vitro immunological and biological activity of four commercially available preparations of human chorionic gonadotropin (hCG) for clinical use (Predalon(R): hCG-A, Primogonyl(R): hCG-B, Choragon(R): hCG-C and Pregnisin(R): hCG-D) were examined. The methods used include immunoassay, cAMP production in Chinese hamster ovary (CHO) cells and structural analysis of hCG with high performance liquid chromatography (HPLC). There were remarkable differences in immunological activities among preparations. hCG-D had the highest concentration of free hCGbeta. In CHO cells, production of cAMP with hCG-C was significantly higher than that with other preparations. Analyses in HPLC showed similar pattern in hCG C and a standard preparation of hCG (CR 123). The highest immunological activity in hCG-A could be attributed to the presence of nicked form of hCG in this preparation. Various hCG components in preparation may explain these differences. PMID- 11935498 TI - [Isolated decrease of haptoglobin during pregnancy: diagnosis by chance or pathological? ]. AB - Haptoglobin is an acute-phase-protein, which is important in many diseases like infections, trauma and neoplasma. An increase in haptoglobin is induced by cytocines like IL-6 und IL-1. The normal range for plasmatic haptoglobin is 50 220 mg/dl. During pregnancy the most likely diagnosis is the HELLP- Syndrome (hemolysis, elevated liver enzymes and low platelets), followed by rare diagnoses like viral hepatitis or favism. We report about a 31-year-old III-gravida 0-para at 31 weeks of gestation with a decrease of haptoglobin over a period of 6 weeks (cut off: < 13 mg/dl) and without any clinical signs of preeclampsia. Liver enzymes were constantly slightly elevated without any progress, other laboratory test results were normal. The patient had a caesarean section at 37 weeks of gestation. Serum haptoglobin returned to normal values within three days after delivery. The reason for the decrease of haptoglobin in our case remains uncertain. Further studies need to focus on the differential diagnosis of a decrease of haptoglobin as well as an isolated decrease of haptoglobin during pregnancy and on the valid ranges of the different haptoglobin subtypes. PMID- 11935501 TI - [Intestinal polyp of the umbilical cord]. AB - The morphology, pathogenesis, complications and differential diagnosis of an intestinal polyp of the umbilical cord are presented. The polyp were detected postnatal on the umbilical cord in an healthy male newborn. The presents of intestinal tissue upon the umbilical cord ist possible about the persistence from remnants of the ductus omphalomesentericus with prolapse and differentiation of the intestinal cells. The ductus omphalomesentericus is a tubular structure, a communication between the developing embryonic gut and the yolk sac, forming during the early embryonic life. Obliteration of the omphalomesenteric duct is usually complete by the 10(th) week of gestation. Various portions of the duct may persist, however, giving rise to polyps, fistulas or cysts of the umbilical cord with potentially dangerous clinical consequences. Other tumors of the umbilical cord are myxoma, angioma and teratoma are differential diagnosis. PMID- 11935500 TI - [Trends of OC use 1980-1999 in a German cohort of women]. AB - INTRODUCTION: There is no doubt, oral contraceptives are a safe, reversible and commonly used method of contraception. Ever use is about 80 % in developed countries and current use of OCs about 40 %. Almost all information about prevalence of OC use came from cross-sectional studies and did not distinguish between high- and low-dose OCs. This paper deals with the prevalence of OC use as it was observed during lifetime of a large cohort of German women by calendar year and estrogen content. METHODS: The prevalence of OC use between 1980 to 1999 was analysed on occasion of an interim analysis of the German cohort study on women's health. This analysis is based on more than 10 000 women with about 390 000 women-years of observation. RESULTS: The proportion of women under the age of 50, that ever used OCs, increased steeply from 1980, i. e. from 62 % (1980) to about 90 % from 1996 onward. The proportion of current users in a given calendar year rose from 44 % (1980) to 49 % (1991-95) and dropped after the "pill crisis 1995". The percentage of current users who used high-estrogen-dose OCs dropped from 32 % (1980) to 5 % (1999). In contrast, the percentage of users of low-dose OCs of the 2nd generation steeply increased as was the percentage of users of the 3rd generation pills, however the latter dropped after 1995. CONCLUSION: The high acceptance of oral contraceptives despite many "pill crises" underlines the continuously high appreciation of their efficacy on the one hand, but underscores also the high responsibility of physicians and industry concerning surveillance or reduction of discussed side effects. PMID- 11935504 TI - The use of biomechanical orthodontic treatment concepts for removable appliances in maxillofacial prosthodontics. Part I: review of prosthesis design principles. AB - Maxillary cuspids are commonly included in ablative block resections in maxillectomy patients. Although the remaining incisors are poor abutments for interim or definitive obturators, they must be used if adequate retention and support is to be achieved. Orthodontic biomechanical concepts for removable appliances offer solutions for the treatment of these patients. The proper application of these concepts can improve retention, support, and force distribution to the anterior teeth and is particularly useful in younger patients when long-term maintenance of teeth is critical. The purpose of this article is to describe the rationale for this approach and its application to interim and definitive Aramany Class I and II obturators. PMID- 11935502 TI - [Uterine myomatosis with senile hematometra in a 101-year-old patient]. AB - A 101-year-old patient presented with uterine myomatosis and bleeding associated with a large hematometra. An abdominal hysterectomy with adnexectomy was performed following an in-depth consultation with the patient and her daughter. The patient was discharged on the 12(th) postoperative day after a complication free course. This case report demonstrates not only the current possibilities in the gynecological treatment of very old patients but also the hidden reserves in the gynecological care ot these women. PMID- 11935505 TI - The history and development of the dental surveyor: part I. AB - The dental surveyor is an indispensable instrument capable of performing a number of prosthodontic procedures. Surprisingly, the technique of surveying casts as part of the treatment planning procedure did not emerge until early in the twentieth century. This article reviews the origins and early use of the dental surveyor. It includes features common to all surveyors as well as the principal uses of these instruments. PMID- 11935506 TI - Effect of dowel space preparation and composite cement thickness on retention of a prefabricated dowel. AB - PURPOSE: This investigation examined what effect cement thickness had on retention of prefabricated endodontic dowels luted with a composite resin cement (Panavia 21 OP; J Morita, Irvine, CA) into canals previously obturated with gutta percha and a eugenol-based sealer. MATERIALS AND METHODS: Sixty-four single rooted teeth were decoronated, filed, cleaned, and sequentially shaped with sizes 2-5 Gates Glidden drills (Dentsply/Maillefer, Tulsa, OK) and 0.12 taper rotary instrumentation. Teeth were then divided into 4 groups of 16 specimens each. All specimens were obturated with gutta percha and a eugenol-based sealer. Dowel space preparation and dowel cementation were completed 1 week after obturation. Ten-millimeter-deep dowel spaces were prepared using dowel drills with 4 different diameters: size 5 Parapost drill (Group 1; Coltene/Whaledent, Mahwah, NJ); size 5.5 Parapost drill (Group 2; Coltene/Whaledent); size 6 Gates Glidden drill (Group 3; Dentsply/Maillefer); size 6 Parapost drill (Group 4; Coltene/Whaledent). Size 5 Paraposts were then cemented with Panavia 21 OP. After 48 hours of storage, specimens were mounted in polyvinyl chloride (PVC) pipe with acrylic, and the dowels were removed in tensile mode using a universal testing machine at 1 mm/min, with data recorded in kilograms. RESULTS: (all values in kilograms) Group 1 (Parapost 5) mean = 15.07, 95% confidence interval (CI) = +/ 6.11; Group 2 (Parapost 5.5) mean = 25.60, 95% CI = +/-7.39; Group 3 (Gates Glidden 6) mean = 43.15, 95% CI = +/-7.81; Group 4 (Parapost 6) mean = 37.75, 95% CI = +/-6.35. Analysis of variance and Bonferroni tests revealed that Group 3 had significantly greater mean retention strength values than Group 1 and Group 2 (p <.05), and that Group 4 had significantly greater mean retention strength values than Group 1 (p <.05). CONCLUSIONS: Paraposts cemented with Panavia 21 OP showed significantly greater retention in oversized dowel spaces compared with dowel spaces prepared with the manufacturers' matched dowel-drill set. PMID- 11935507 TI - In vitro evaluation of the retention of composite fiber and stainless steel posts. AB - PURPOSE: This study compared the tensile retentive strength of composite fiber reinforced dowels luted with a resin cement to stainless steel dowels luted with zinc phosphate cement. MATERIALS AND METHODS: The crowns of 40 extracted human mandibular premolars were removed with a separating disc 1-mm coronal to the cementoenamel junction. The teeth were randomly divided into 4 groups (n = 10). A post space was prepared in each specimen to a depth of 9 mm, using the drill supplied by the respective manufacturer. For the stainless steel post group, 1.25 mm-diameter posts were cemented with zinc phosphate cement. For the composite fiber reinforced groups, posts with diameters of 1.00, 1.25, and 1.50 mm were luted with a Bisphenol A-Glycidyl Methacrylate (BIS-GMA) resin bonding system (Jeneric/Pentron, Wallingford, CT) according to manufacturer specifications. The specimens were stored in a sealed container with a moist environment for 24 hours, placed in a fixture in an 801 Materials Test Systems (MTS) machine (MTS Systems Corp, Minneapolis, MN), and loaded in tension at a rate of 5.0 mm/min until failure. Differences among the 4 groups were determined using a one-way analysis of variance and Tukey-B post-hoc tests (alpha = 0.05). RESULTS: The mean loads to failure ranged from 43.9 +/- 10.4 kg for the stainless steel dowel group to 19.9 +/- 5.7 kg for the composite fiber-reinforced 1.00-mm-diameter group. The stainless steel post luted with zinc phosphate cement provided significantly greater tensile resistance than all composite fiber dowel groups. The retention of the 1.00-mm composite fiber-reinforced post was significantly less than the remaining groups. CONCLUSIONS: Under the conditions of this study, the stainless steel dowel luted with zinc phosphate cement provided significantly greater retention. PMID- 11935508 TI - Finite element analysis on dental implant-supported prostheses without passive fit. AB - PURPOSE: The purpose of this study is to use finite element analysis to investigate the effect of misfit prostheses, cantilever prostheses, and various occlusal forces on the stress distribution in the prostheses, implant components, and surrounding bone. MATERIALS AND METHODS: Two 3-dimensional finite element models were constructed: (1) a 2-implant-supported, 2-unit fixed partial denture and (2) a 2-implant-supported, 2-unit fixed partial denture with a distal cantilever. Variations of the standard finite element models were made by placing a 111-microm gap between the gold cylinder on either the mesial or distal implant. The effects of load of 100 N were tested on all models. Subsequently, loads of 50 N, 200 N, and 300 N were evaluated on the cantilever model. RESULTS: When the gap was positioned near to the applied force, the stress in both models increased significantly in the implant components and surrounding bone. The stress increase in each component ranged from 8% to 64% in the non-cantilever models and 43% to 85% in the cantilever models. The greatest stress was found in the distal gold screw. The effect of the gap was clearly shown by the pattern of stress distribution in both models. Additionally, the presence of a cantilever and excessive occlusal force amplified the effect of prosthesis misfit. CONCLUSIONS: Prosthesis misfit influenced the pattern and magnitude of stress distribution in the prosthesis, implant components, and surrounding bone, and the presence of the cantilever or greater occlusal force amplified the effect of misfit. PMID- 11935509 TI - A maxillary implant overdenture that utilizes angle-correcting abutments. AB - Placement of dental implants to enhance complete denture stability and retention is a well-accepted treatment modality. Most implant overdenture abutments and related components require parallelism within approximately 10 degrees to function properly. Non-parallel implant placement as a result of flared maxillary bone may preclude the use of conventional implant overdenture abutments, requiring the use of costly custom abutments or bar prostheses. A case report is presented that illustrates the use of a new, 2-piece, angled overdenture abutment to accommodate non-parallel implant placement. PMID- 11935510 TI - A provisional fixed partial denture that simulates gingival tissue at the pontic site defect. AB - A technique is presented for the fabrication of an esthetic, provisional fixed partial denture that compensates for a pontic-site ridge defect. This provisional restoration enables both the dentist and the patient to evaluate whether this prosthetic approach will adequately camouflage the pontic-site defect or whether surgical correction of the pontic site should also be considered. PMID- 11935511 TI - Installing magnetic keepers using LASER welding. AB - Magnetic overdenture attachments can provide useful denture retention. A technique is proposed that involves installing the magnetic keeper in the overdenture abutment coping using laser welding. Compared with the conventional cast-to technique, the new technique provides a suitable contour to the abutment coping with the magnetic keeper and ensures proper mounting in the abutment coping, installing the magnetic keeper easily and effectively. PMID- 11935512 TI - The history of articulators: from facebows to the gnathograph, a brief history of early devices developed for recording condylar movement: part II. PMID- 11935514 TI - Undertreating pain may be elder abuse. PMID- 11935515 TI - FDA backs cancer drug for treating MS. PMID- 11935517 TI - Some HIV patients delaying or forgoing treatment. PMID- 11935519 TI - Surviving the merger, the acquisition, or shall we call it a takeover? PMID- 11935520 TI - Integrating CAM into traditional health care. PMID- 11935521 TI - State and federal regulation of health care: friend or foe? PMID- 11935522 TI - Proving the value of case management. PMID- 11935523 TI - Custom equipment: why does it take so long? PMID- 11935524 TI - CCM certification questions. PMID- 11935525 TI - The effective integration of utilization and case management. PMID- 11935526 TI - The importance of patient education materials. PMID- 11935527 TI - Proposed practice guidelines for excellence in life care planning. PMID- 11935528 TI - The Americans with Disabilities Act: implications of Supreme Court decisions for case managers. Part II. PMID- 11935529 TI - Could a process improvement program improve your quality assurance. PMID- 11935531 TI - Playing the political game. PMID- 11935530 TI - Caring for Houston's Vietnamese population. PMID- 11935532 TI - Advanced nursing practice in Kentucky. PMID- 11935533 TI - Severe head injury in children: a case study. PMID- 11935534 TI - Why are we waiting. PMID- 11935535 TI - Thriving or surviving? Challenging the boundaries of ENP practice. PMID- 11935536 TI - A special breed of nurse. PMID- 11935537 TI - Resuscitation training in Uganda. PMID- 11935538 TI - Pain assessment tools. PMID- 11935539 TI - Gut decontamination and poisoning. PMID- 11935540 TI - The future of A&E with telemedicine. PMID- 11935541 TI - Learning on the job accredited work based learning. PMID- 11935542 TI - The health psychology of asthma. PMID- 11935543 TI - An investigation into the assessment and management of pain by triage nurses in Greater London A&E departments. PMID- 11935544 TI - Using telemedicine for remote trauma medicine. What the public thinks. PMID- 11935545 TI - Looking after children. PMID- 11935546 TI - Developing paediatric nursing skills. PMID- 11935547 TI - Ethical implications of achieving pain management through advocacy. PMID- 11935548 TI - Organisation of care for older people in A&E. PMID- 11935549 TI - Care of the ventilated patient in A&E. PMID- 11935550 TI - Pre-hospital trauma awareness for nurses. PMID- 11935551 TI - An organic concept in nursing. PMID- 11935552 TI - Emergency nursing: a New Zealand perspective. PMID- 11935553 TI - Emergency nurse practitioners should manage major cases. PMID- 11935554 TI - Problems with eating fungi. PMID- 11935555 TI - Triage: a retrospective evaluation. PMID- 11935556 TI - Why do A&E nurses fear using their advanced life support skills? PMID- 11935557 TI - Emergency nurse practitioner's documentation: development of an audit tool. PMID- 11935558 TI - Nursing in cyberspace. PMID- 11935562 TI - A plan for spreading the word about case managers. PMID- 11935563 TI - PhDx e-SYSTEMS offers Web-based efficiency software. PMID- 11935564 TI - Complementary and alternative medicine: the public wants more. PMID- 11935565 TI - Best practices and outcomes. PMID- 11935566 TI - Long-term care insurance and case management--the last frontier. PMID- 11935567 TI - Life care planning: a tool for elder care management. PMID- 11935568 TI - Case management's role in a Communist health care system. PMID- 11935569 TI - The Americans with Disabilities Act: implications of Supreme Court decisions for case managers. AB - The Supreme Court interpreted the application of the ADA to be based on an individualized evaluation of a person's circumstances. A full, objective vocational evaluation continues to be a valuable tool for the employee, insurance company, employer, attorney, and judicial system. Case managers must continually evaluate an individual's potential in light of his or her circumstances, not just the medical diagnosis. PMID- 11935570 TI - Personal care management at the heart of social HMOs. PMID- 11935571 TI - Case managers in the hospital workplace--a light-hearted view at century's end. PMID- 11935572 TI - Are you a burnout candidate? PMID- 11935580 TI - Your invited guests are at the door, or who needs accreditation? PMID- 11935582 TI - ClickON managed care suite for optimum workflow. PMID- 11935581 TI - Screening for use of CAM therapies. PMID- 11935583 TI - AIDS disease management programs. PMID- 11935584 TI - Case managers and the Internet--a roadmap for e-case management. PMID- 11935585 TI - Tell us what you really think! Case managers weigh in on the profession. PMID- 11935588 TI - The latest trends in the pharmaceutical market. PMID- 11935586 TI - The state of affairs in Alzheimer disease. PMID- 11935590 TI - Culture of silence. PMID- 11935591 TI - Hospital management of poisoning in victims suffering from smoke inhalation. PMID- 11935592 TI - Climate of care and nurses' attitudes towards AIDS in the emergency department. PMID- 11935593 TI - Ongoing debate about ENPs. Degree level education vs experience in practice. PMID- 11935594 TI - Recognising the survivors of torture. PMID- 11935605 TI - Get with the program and start training with a plan. PMID- 11935606 TI - HMOs and alternative care. PMID- 11935607 TI - e-MEDx, Inc.'s MedAdvice 2001. PMID- 11935608 TI - New standards, guide, programs on horizon. PMID- 11935609 TI - Acute care innovations and outcomes in the military medical system. PMID- 11935610 TI - PACE programs. Part II. PMID- 11935611 TI - Posttraumatic stress disorder and the injured worker. Part II. PMID- 11935612 TI - The Americans with Disabilities Act: implications of Supreme Court decisions for case managers. Part IV. PMID- 11935613 TI - Crisis in case management: the graying of America. PMID- 11935614 TI - Life care planning practice: external influences. PMID- 11935616 TI - The third world is here. PMID- 11935615 TI - The influence of special needs trusts on case management practice. PMID- 11935617 TI - Minor injury services--the present state. PMID- 11935618 TI - Sulphonylureas and metformin overdose: clinical features and management. PMID- 11935619 TI - Caring for people with Down syndrome in A&E. PMID- 11935621 TI - New guidance set for A&E service provision. PMID- 11935620 TI - Shock: types, classifications and explorations of their physiological effects. PMID- 11935622 TI - Recognising and assessing victims of torture in emergency departments. PMID- 11935624 TI - Mutual of Omaha lifts AIDS coverage cap. PMID- 11935632 TI - The challenge: hire the right people for the right reasons. PMID- 11935634 TI - WebRN: practice-specific Internet resource. PMID- 11935633 TI - Integrating CAM into traditional health care. PMID- 11935635 TI - Return-to-work interventions and outcomes. PMID- 11935636 TI - PACE programs. Part 1. PMID- 11935637 TI - New genes. PMID- 11935638 TI - When the case manager becomes the patient. PMID- 11935639 TI - The research behind the CCM examination. Interview by Patricia McCollom. PMID- 11935640 TI - Posttraumatic stress disorder and the injured worker. Part I. AB - PTSD occurs in a small but significant number of people who experience life threatening events. When a disorder occurs, it can have severe and long-lasting effects on a person's ability to work and function effectively. Particularly effective treatment interventions give patients the opportunity to synthesize and integrate their traumatic memories. Such strategies increase their ability to more narrowly limit the traumatic event and broaden personal, occupational, and social experiences that are healthy and positive. The results is that, although the traumatic experience may be recalled, it is seen as a single autobiographical event in a rich chain of personal experiences. PMID- 11935641 TI - The road ahead--managing a spinal cord injury. PMID- 11935642 TI - Maximizing the effectiveness of case management service delivery. PMID- 11935643 TI - The Americans with Disabilities Act: implications of Supreme Court decisions for case managers. Part III. PMID- 11935644 TI - The challenges of providing case management in a Medicaid environment. PMID- 11935645 TI - A cost-effective model of community case management. PMID- 11935646 TI - Case management in psychiatric disability. PMID- 11935647 TI - My neck hurts: an audit of self presenting patients with neck injury. PMID- 11935648 TI - Pesticide poisoning: rodenticides. PMID- 11935649 TI - Farm accidents: their causes and the development of a nurse led accident prevention strategy. AB - The following key facts emerge which are relevant to an accident prevention strategy. HIGH RISK TIMES: Just before lunch and late afternoon. HIGH RISK PLACES: Those places which are most familiar to the farmer, his/her own farm, especially the yard and farm buildings. HIGH RISK ACTIVITY: Working at heights (falls) or with moving objects, tractors and cattle. HIGH RISK PEOPLE: Younger men, especially when not working on their own farm or older farmers working with livestock. The majority of these accidents could have been prevented with the correct use of protective clothing or taking more care. Once an accident has happened, the first aid response is likely to be inadequate. The project team are now working on a strategy to persuade farmers that accidents could happen to them personally and of the seriousness of the consequences if they do. This will be coupled with producing accident prevention materials outlining simple precautions that can be taken to prevent accidents. The final stage is to disseminate these materials through a range of channels involving both NHS, farming and school networks. That is the next stage of the project. PMID- 11935650 TI - Evaluating the development of an accredited work based learning (AWBL) scheme for A&E nurses. PMID- 11935651 TI - Life as an expedition medic in Chile. PMID- 11935652 TI - [Minimal residual disease in solid cancer. Prognostic value and therapeutic consequences]. AB - Minimal residual disease (MRD) is one of the issues in solid carcinomas which is still discussed controversially as to its prognostic value and therapeutic implications. In the past, the evidence for MRD was especially given by immunocytochemistry for epithelial cytokeratin markers which were able to detect single disseminated tumor cells within the mesenchymal background of the bone marrow at sufficient sensitivity and specificity. However, for the perioperative detection of disseminated tumor cells, numerous studies have given controversial results regarding the prognostic relevance of these cells. Some other studies investigating the postoperative development of disseminated tumor cells in the bone marrow after curative tumor resection observed substantial changes in status over time, suggesting that microenvironment interactions and phenotypic characteristics of these cells rather than the mere evidence of MRD will be able to indicate the stage of systemic disease which is of metastatic and prognostic relevance. Currently, new molecular biology techniques are applied to achieve the objective of MRD phenotyping which has good chances of providing new models for a new molecular tumor staging. Based on this development, targeting biologically relevant disseminated tumor cells will most likely become an important issue in carcinoma therapy, and first therapeutic studies are under way. However, at the present time, the mere screening for disseminated tumor cells must still be seen as a very helpful diagnostic tool for monitoring MRD especially in the setting of follow-up, rather than being able to be a key essential in therapeutic applications. PMID- 11935653 TI - [Abscesses, endophthalmitis and retinopathy as "initial manifestations" of type 2 diabetes]. AB - The case of a 42 year old female patient is described, who presented with abscesses in the lung, the left kidney, the perspiratory glands of the axillae and endophthalmitis as well. Staphylococcus aureus could be isolated from sputum, urine, bronchoalveolar lavage and excisates of the abscesses in the right axilla. After long-term antibiotic and antimycotic treatment, restitutio ad integrum could be achieved. Since the further course excluded underlying malignancy, the septic course of Staphylococcus aureus infection must be regarded as an effect of immunosuppression in long-standing and untreated diabetes mellitus. PMID- 11935654 TI - [Effectiveness of zopiclone in disorders of initiating and maintaining sleep. Results of a drug monitoring study in 811 general practices]. AB - BACKGROUND AND METHOD: In a drug monitoring study with 811 participating general practitioners, safety and tolerability of zopiclone were studied in 2416 patients with disorders of initiating and maintaining sleep. RESULTS: In general, zopiclone was efficient in all forms of insomnia; the subjective sleep duration was prolonged for 2 hours on average. Patients without somatic complaints or comorbidity showed the greatest benefit. Daytime well-being and vigilance were in general not impaired. Drug related adverse events occurred rarely; the great majority of the participating physicians rated the treatment with zopiclone as efficient and acceptable. CONCLUSION: In this drug monitoring study, zopiclone proved an efficient hypnotic for the treatment of disorders of initiating and maintaining sleep. PMID- 11935655 TI - [Do old persons wish to die? Suicidal tendencies and weariness of life in old age. Results of the Berlin Aging Study (BASE)]. AB - On the basis of the results of the Berlin Aging Study (BASE), some 15% of persons aged 70 years and older confess to being tired of life, 5% express a wish to die, and 0.5-1% entertain thoughts of suicide. Women express a wish to die or have thoughts of suicide approximately 1.5 to 3 times more frequently than men. Depending on the severity of the suicidal tendency, more than 80% of such patients, and all of these entertaining thoughts of suicide, suffer from acute psychiatric disorders. Death wishes and thoughts of suicide must therefore always prompt an intensive psychiatric diagnostic work-up, and initiation of suitable and rigorous treatment, and must not be mistaken for the expression of understandable attitudes towards aging. PMID- 11935656 TI - [Diabetic dyslipoproteinemia: physiopathological bases and treatment prospects]. AB - Dyslipoproteinemia associated with type 2 diabetes comprises hypertriglyceridemia caused by reduced insulin sensitivity, and consequently, low HDL levels and an increase in the proportion of small dense LDL particles. In addition, in both type 1 and 2 diabetes glycated LDL is formed in the presence of high plasma glucose levels. These lipoprotein disorders are all atherogenic and are responsible for the distinctly increased risk for cardiovascular disease in diabetics. Intensive glucose-lowering measures result in lower rates of micro- and macro-angiopathies in both types of diabetes. The benefit of additional lipid lowering measures has not yet been confirmed by appropriate investigations. However, subgroup analyses from two large intervention trials do demonstrate that mortality from coronary heart disease may be substantially reduced. LDL cholesterol levels in diabetics should not exceed 115 mg/dl (3 mmol/l), and fasting triglycerides should be lower than 180 mg/dL (2 mmol/l). PMID- 11935657 TI - [Percutaneous placement of feeding tubes for long-term nutrition and decompression. Indications, techniques and complications]. AB - In the recent past, PEG/PEJ has found worldwide acceptance. The uncomplicated nature of the method has resulted in a considerable increase in endoscopic gastrostomies. Provided that the method is properly applied, and appropriate aftercare is offered, complications rarely occur. In principle, the indications are chronic diseases requiring tube feeding for longer than one month, such as cachexia in patients with cancer or stenotic tumors of the mouth and throat, or of the upper gastrointestinal tract. PMID- 11935658 TI - [Misdiagnosis of first onset schizophrenia in adolescents]. AB - The intensity of the symptomatology of schizophrenia in adolescents is essentially the same as in adults. As a rule schizophrenic psychoses in adolescence are first manifestations, and other psychiatric disorders have to be ruled out. The classical differential diagnoses include: exogenous psychoses, in particular drug-induced; affective psychoses; pubertal crises; personality disorders. Special early-onset features arise because drastic behaviour of another nature is occasionally misinterpreted or--more commonly the case--early schizophrenia in adolescents is not recognised and treated in time because of the uncharacteristic initial symptoms. PMID- 11935659 TI - [Molecular biology of cystinuria]. AB - Cystine stones belong to the kidney stones diseases which are difficult to manage due to the high rate of recidivation and the necessary surgical procedures. Cystinuria is an autosomal recessive defect of the transport of cystine and the dibasic amino acids in the proximal kidney tube. Recently, two subunits of a renal cystine transporter have been identified. Mutations in these genes have been shown to lead to the cystinuria phenotype. Genetic and functional analyses have helped to modify the classification of cystinuria which had previously been exclusively based on biochemical data. Furthermore, first steps towards a molecular genetic testing have been carried out replacing the so far diagnostic procedure which are stressing for the patients. The results of these testings make an individual therapy possible. PMID- 11935660 TI - [A therapeutic approach in Burkitt's lymphoma. 2 case reports from the Hospital Central da Beira/Mozambique]. AB - Burkitt's lymphoma is the most common malignant tumor encountered in children (age peak 3 to 8 years) living in areas with endemic malaria tropica. In the Hospital Central da Beira/Mozambique, two children with Burkitt's lymphoma were treated with cyclophosphamide. During the period covered by this report--August 1 to September 30, 2000--two of the required four treatment cycles were carried out, and both children experienced a remission. PMID- 11935661 TI - [NISAS-2000: The "Nationwide Insomnia Screening and Awareness Study". Prevalence and interventions in primary care]. AB - AIM: To estimate the point prevalence of insomnia, recognition and prescription behavior in primary care. METHODS: Nationwide sample of 539 primary care settings along with their characterization (stage 1). Standardized assessment of all attenders (N = 19.155 patients) on the NISAS target day using a sleep questionnaire (PSQI) and additional questions to cover psychosocial and additional clinical variables. All patients were evaluated by the primary care doctors using a standardized clinical appraisal questionnaire, including a CGI rating. RESULTS: Prevalence insomnia according to DSM-IV was 26.5%. Recognition of presence of any clinically significant sleep disorder was 72%, recognition of insomnia was poor 54.3%. 85.6% of insomnia patients were rated as chronic. Close to 50% of all insomnia cases did not receive a specific insomnia therapy. Herbals, followed by hypnotics and sedatives and antidepressants were the three most frequent treatments applied, psychotherapy was only seldomly indicated. DISCUSSION: NISAS provides for the first time nationally representative estimates of interventions for insomnia in primary care. The relatively low treatment rates and the high proportion of chronic patients receiving longterm prescription of benzodiazepines seem to be critical. Priorities for future agenda to improve this situation are discussed. PMID- 11935662 TI - [Generalized anxiety disorders (GAD)--a neglected illness? Background und aims of the GAD-P study]. AB - In the past Generalized anxiety disorder (GAD)--previously classified as anxiety neurosis--was regarded as not being a separate diagnostic entity. On the basis of new explicit criteria for GAD in the 90ies, GAD-specific pharmacological (i.e. SNRI) and psychological treatments with improved efficacy have become available. The Generalized Anxiety and Depression in Primary care study (GAD-P) investigates the prevalence of GAD in primary care settings and evaluates the patterns of care provided. Aims, methods and findings of the GAD-P study are described in this supplement. PMID- 11935663 TI - [Do primary care physicians know generalized anxiety disorders? Estimations of prevalence, attitudes and interventions]. AB - This paper examines attitudes and characteristics of service providers for generalized anxiety disorders (GAD) in primary care. Based on a nationwide sample of 558 primary care doctors, the study found: 56.9% of all primary care doctors consider GAD to be an independent disorder, younger doctors more frequently than older ones. On average, they see on average at least 2.3 GAD definite patients a day. 27.4% indicate that they treat GAD differently than they treat depression. Doctors with frequent postgraduate courses on either depression or GAD reveal more favourable competence ratings with regard to recognition/diagnoses and therapy and refer such patients less frequently. Suggested treatments for GAD, however, are diffuse and by and large inconsistent with current research knowledge. Overall these findings suggest that the majority of primary care doctors view GAD as a prevalent independent disorder with special treatment needs. PMID- 11935664 TI - [Prevalence and correlates of generalized anxiety disorders in primary care]. AB - AIM: The investigators evaluated the (target-day) point prevalence and correlates of generalized anxiety and depression in German primary care practices. METHOD: The study used a representative national sample of more than 500 primary care practices. On a target day, all primary care patients were examined by questionnaires (GAS-Q and DSQ). RESULTS: Conservatively estimated, 5.3% of all primary care patients met criteria for generalized anxiety disorder (GAD). Similar prevalence estimations were found for Major Depression. Only 30% of all GAD patients met criteria for depression cross-sectionally. Patients with GAD (whether comorbid with depression or not) were more frequently female, reported an onset of GAD mostly between ages of 25 and 45 years, had increased rates of disability and impairment in major social roles and high utilization rates of all health care resources. They also revealed a predominantly chronic course of illness over a period of several years and an increased suicide rate. The presence of physical illness increased rates of handicap/impairment days in the past month. Associated negative outcomes were found to be similar to those observed in depressive disorders. Patients with GAD reported significantly more frequently sleep, anxiety and depressive problems as primary reasons for contact. CONCLUSION: The GAD-P study highlights the significance of GAD in primary care as a condition that occurs at about the same frequency as major depression. Further, the relatively high proportion of pure GAD suggests that the frequency of depression-comorbidity of GAD has previously been overestimated. PMID- 11935665 TI - [When and how well does the family physician recognize generalized anxiety disorders and depressions?]. AB - BACKGROUND: Recognition rates for generalized anxiety disorder (GAS) and depression in primary care and its predictors are reported. METHODS: Based on the results of GAD-P study screening questionnaires, the investigators evaluated how many patients were correctly classified by primary care physicians a) as cases with mental disorders and b) as specific disorder cases. Socio-demographic and illness history variables of patients as well as features of physicians were analyzed as predictors of recognition by regression analyzes. RESULTS: Physicians recognized a mental disorder in more than two thirds of cases with GAS; rates were even higher in patients with comorbid anxiety and depression (85.4%). Recognition of the specific disorder was conspicuously worse in patients with GAS (34.4%) in comparison with patients with depression (64.3%). Only the variables of patients' demographic status and illness (e.g. higher age or, negatively, incident GAS) predicted recognition of a mental disorder. CONCLUSION: Generalized anxiety is often recognized as a mental disorder but seldom as the specific diagnosis of GAS. This is especially true for patients without a history of mental disorders (incident cases). Suggestions for a better recognition are discussed in the light of specific psychopharmacologic and psychotherapeutic treatment demands. The risks of chronicity of this disorder as well as the ascertained predictors of good recognition are also discussed. PMID- 11935666 TI - [Family physician's interventions and prescribing practice. Results of the GAD-P study]. AB - More than two thirds of all GAD patients are recognized as cases with mental disorders, only one third is correctly diagnosed. The paper shows that this has significant implications. 36%--as compared to 23% of those with major depression- receive no intervention. Of those recognized at least as a case the majority is treated by the GP, 9% are only referred to specialists, in addition to another 20% that are treated by the GP and referred as well. Almost all patients receive medication, however, only few medications that match scientific guidelines for GAD-specific treatments, namely SNRI, behavioural psychotherapy or SSRI. Also the high degree of comedication as well as high prescription rates for sedatives and phytotonics needs highlightening. The findings overall reveal an unsatisfactory picture of current treatment strategies for GAD patients in primary--especially if compared to depression. Treatments of first choice, SNRI (Venlafaxine SR) and behavioural psychotherapy are prescribed to only the minority of GAD sufferers. PMID- 11935667 TI - [Problems and deficiencies in family physician's management of generalized anxiety disorders. Results of the GAD-P study and priorities for an improved care]. AB - The core findings of the GAD-P study (Generalized anxiety and depression in primary care) are summarized and measures to improve the quality of care of patients with generalized anxiety disorders are discussed. In addition to the identification of core research deficits the paper emphasizes the standard use of time-efficient diagnostic screening instruments, because urgently needed improved recognition and diagnosis is the prerequisite for appropriate treatment. Further the role of the media to combat stigma processes, as well as patient education materials to improve compliance and to enhance treatment outcome effects for this neglected disorder that frequently runs a chronic course are highlighted. PMID- 11935668 TI - ["Generalized anxiety and depression in primary care" (GAD-P). Study design and methods]. AB - The aim of the study is to develop a comprehensive clinical-epidemiological description of the prevalence of generalized anxiety disorders and depression among primary care patients along with an assessment of physicians recognition rates and prescription behaviour. The paper describes methods and design of the study and provides background information on the sampling process and instruments used as well as characteristics of doctors and patients. The study is based on a nationally representative sample of 558 primary care physicians and over 20,000 patients, who attended physicians' offices on the target day. The first stage of study involved a comprehensive description of the physicians characteristics in terms of psychosocial qualification, and provider aspects as well as attitudes towards GAD and depression. In the second stage, all the patients completed a diagnostic screening questionnaire for GAD and depression. In the third stage all patients were characterized by their physicians in terms of their diagnostic status and their past and current interventions. PMID- 11935669 TI - [Transcranial color-coded Doppler sonography: getting started]. AB - Transcranial color Doppler sonography, in contrary to conventional transcranial Doppler ultrasound, allows outlining of the parenchymal structures and visualization of the vessel examined. Angle-corrected blood velocities can be obtained which are closer to in-situ values. In this educational assay, we demonstrate cerebral vascular anatomy using 3-D MR angiographic images of the major cerebral arteries to show their relationship to neighboring structures, the acoustic window, and the ultrasound beam. We describe principles of the examination, and discuss the different types of physiological flow velocity spectra. We show how to identify with color Doppler ultrasonography the main intracranial arterial trunks, and how to avoid pitfalls caused by anatomic variability of their course and location. We draw attention on significant age and sex dependency of blood flow parameters which makes standardization of obtained transcranial Doppler results indispensable for clinical valuable concluding. PMID- 11935670 TI - [Correlation of expression of progesterone receptors with histopathological type and grade of malignancy of cerebral neoplasms]. AB - Steroid receptors play an important role in homeostasis and in proliferation of neoplastic cells as well. Biological effect of steroid hormone on tissues depends on intracellular concentration of its receptor. The binding hormone with its receptor activates adequate genes in cellular nuclei and cell proliferation. The mutation of genes codifying of receptor protein may cause its structural modification or changes in its activity and lead to neoplastic transformation. The aim of our work was the estimation of progesterone receptors (PR) concentration in cerebral neoplasma and correlation with histopathological type and grade of malignancy of cerebral tumours. We examined 89 patients, among them 28 with meningioma, 43 with glioma and 18 with metastatic tumour. The concentration of PR was evaluated using radioreceptor assay. Our results show statistically significant correlation between expression of PR and type and malignancy of neoplastic tumours. It was especially seen in meningioma and the highest concentration of PR were in I grade tumours. PMID- 11935671 TI - [Syringohydromyelia--coexistence or complication following removal of meningioma of forth ventricle]. AB - Numerous theories have been presented to explain the pathogenesis of syringohydromyelia. Among these is Gardner's hydrodynamic theory with later modifications. Syrinx is usually located in cervical cord and is often associated with Arnold-Chiari, Dandy-Walker malformation or other anomalies of cranio cervical region. Other theories suggest that syringohydromyelia may be secondary to arachnoiditis, neoplasms or spinal cord trauma. The distal location of the syrinx is very often associated with occult spinal dysraphic lesion. The authors present the case of 28 years old woman operated on meningioma of fourth ventricle. One and a half year later syrinx on Th11-Th12 and coexistent arachnoiditis were recognized. They stress diagnostic and therapeutic difficulties in this case and try to explain of etiology of syringohydromyelia. PMID- 11935672 TI - [The treatment of cerebral abscesses in children--the clinical experiences of neurosurgical clinic of PMMH in Lodz]. AB - In our study we made attempt to assess which clinical criteria are essential for choosing optimal treatment for children suffering from cerebral abscesses. We observed the clinical results obtained at our Department from treatment of infections of the central nervous system (CNS) in children and analysed them with respect to applied therapy and clinical conditions. Between 1990 to 2000, 45 children aged 3 days to 15 years (mean 1.34 years, standard deviation 3.23 years) with infections of the CNS were treated at our Department. Within this group, 26 patients were diagnosed with cerebral abscesses, 1 with epidural empyema, 3 with subdural empyema and 15 with pyocephalus. 42 patients were operated. In 3 patients a conservative treatment was introduced at the moment when the abscesses were in the initial stage of organisation; in each case after a complete cycle of antibiotic therapy a regression of lesion was noted. The most frequently performed--in 19 patients--surgical procedure was the radical excision of the abscess by craniotomy. In 2 cases a re-operation was necessary. The highest rate of re-operations was noted in the group of children treated by external drainage; within this group of 18 patients only one 3-month-old girl did not require an additional surgical intervention. Neuroendoscopic techniques were used in 7 patients with compound hydrocephalus. Out of 45 treated children, 3 died from encephalitis, the primary reason being the infection of the ventriculo-peritoneal shunt. All children were assessed according to the Glasgow Outcome Scale and the results were significantly worse in those under 3 years in age. We suggest that in children above 3 years of age with mature abscesses of the central nervous system an operative evacuation of the abscesses by craniotomy should be performed. PMID- 11935673 TI - [Endothelin in the plasma and cerebrospinal fluid of patients after head injury]. AB - Endothelin (ET) is identified as a potent vasoconstrictor peptide. It can cause a cerebral vasospam after subarachnoid hemorrhage. Its long-lasting vasoconstricting activity has been well documented. The role of ET in response to head injury is not clear. ET can participate in astrocyte activation and oxidative stress after trauma. The aim of this presentation was to investigate ET in the plasma and cerebrospinal fluid of patients after head injury and estimation of relation between the ET csf level and clinical condition of the patients and also relation of ET csf level and changes in the CT-scan and usefulness of ET as independent prognostic factor. We examined 30 patients at Day 1 after trauma (28 male, 2 female, aged 19-34 y, mean 30, 4, GCS ranged 8-15). CT examination showed post-traumatic changes in 19 cases (11 with brain edema only, 8 with brain contusion). We measured ET by immunoenzymatic method using standard kits (Biomedica). The ET plasma level reached 1.12 +/- 0.63 fmol/ml (in the control group--0.44 +/- 0.19 fmol/ml) and c ET csf level 1.03 +/- 0.49 (in the control group--0.07 +/- 0.09 fmol/ml). W confirmed the correlation between the ET level and patients consciousness (by GCS) (p < 0.01). The ET csf level was highest in the patients with brain contusion in CT-scan (p < 0.02). Using multivariate analyses, we showed that ET can be used as a prognostic factor of results of treatment (p < 0.02). CONCLUSIONS: The ET level is elevated in the plasma of patients after head injury. ET is also present and strongly elevated in the cerebrospinal fluid of the same patients. The ET level in cerebrospinal fluid is associated with clinical condition of patients and elevated in cases with brain edema and structural brain damage. The ET level in cerebrospinal fluid is related to post-traumatic changes of the brain such as oedema and structural damage present in the CT-scan. ET can be used as a prognostic factor of results of treatment. PMID- 11935674 TI - [Von Willebrand factor in patients with subarachnoid hemorrhage]. AB - Von Willebrand factor (vWF) plays an important role in the coagulation system. It affects platelet aggregation in the place of vessel endothelium damage. The importance of vWF in cerebrovascular disease is not clear. The aim of this presentation was to evaluate the vWF level in the plasma and cerebrospinal fluid of patients after subarachnoid hemorrhage including cases with cerebral vasospasm and cerebral infarct. We investigated 66 patients (38 persons graded I-III H&H and 28 persons graded IV H&H). The control group consists of 8 subjects. VWF was measured by ELISA method using standard kit Asserachrom (Boehringer). The vWF level in the plasma reached 288.81 +/- 99.66% (in the control group, 98.84 +/- 14.53%) without significant differences regarding clinical condition of patients. In the cerebrospinal fluid the vWF level was significantly different in I-III H&H patients and in IV H&H patients (1.21 +/- 0.52% and 9.18 +/- 7.58%, respectively, p < 0.001) and in the control group (0.13 +/- 0.33%). Our data indicate that there is correlation (Pearson, p < 0.01) between the level of vWF and neurological condition (GCS) of patients. There is also correlation (p < 0.01) between the level of VWF and the presence of clinical disorders (cerebral vasospasm, delayed neurological deficit (DIND) and cerebral infarct). Based on multivariate analysis, we confirmed that vWF is an independent prognostic factor of cerebral vasospasm (p < 0.01) and ischemic complications (p < 0.02). CONCLUSIONS: vWF is elevated in the plasma of patients after subarachnoid hemorrhage in early stage of the disease. VWF is present in the cerebrospinal fluid of patients with SAH and its level is higher in patients with poor neurological condition. The VWF elevation in csf is correlated with clinical condition of patients and also the presence of cerebral vasospasm and cerebral infarct. VWF can be used as an independent prognostic factor of cerebral vasospasm and ischemic complications. PMID- 11935675 TI - [Clinical application of proton magnetic resonance spectroscopy for differential diagnosis of pediatric posterior fossa tumors]. AB - We report here on a correlation between proton magnetic resonance spectroscopy (MRS) spectra obtained in children with posterior fossa tumors and tumor histology and grading. Twenty-six children (age 1-16) were investigated before surgery by using single-voxel proton MRS. All examination were performed on a 1.5 T MR scanner by using single-voxel (8 cm3) with PRESS sequence (TR 1600 ms, TE 270 ms, NEX 256). Spectra of N-acetylaspartate (NAA), choline containing compounds (Cho), creatine and phosphocreatine (Cr) and lactate (Lac) were evaluated. Absolute concentrations of the metabolites were measured and their ratios were calculated. Correlation between these and tumor histology and grading were then determined. Concentration of Cho and Lac, and Cho/NAA ratio were the major statistically significant parameters for discrimination between benign (WHO grade I and II) and malignant tumors (WHO grade III and IV), in particular between pilocytic astrocytomas and medulloblastomas. Discrimination between individual histological types within malignant and benign tumor groups was not possible. Proton MRS of pediatric posterior fossa tumors seems to be helpful in prediction of tumor grading and histology. Specific character of the examination requires establishing of the individual standards for every MR scanner. PMID- 11935676 TI - [Application of the insulating material teflon--pladgets (Genzyme-USA) during the microvascular decompression]. AB - Various isolation materials have been used in the treatment of trigeminal neuralgia due to neuro-vascular conflict. Surgical treatment (MVD) currently utilizes for isolation soft teflon pladgets, which are, after appropriate preparation, placed in the form of wool between the conflicted structures. The material was used in 20 surgical procedures. The definite advantage of this material as compared to others is the possibility to position it correctly between the blood vessel and the nerve without additional fixation with e.g. a tissue adhesive. Observations indicate that this is the best isolation material of those used so far in MVD procedures. PMID- 11935677 TI - [Assessment of p53 dependent apoptosis in glia-derived tumors of the brain]. AB - There are conflicting reports in connection with the association of the p53 tumour suppressor gene mutation with the clinical and histopathological progression of gliomas. Glia-derived neoplasms frequently show mutational changes in the p53 gene which result in enhancement of tumorigenesis. The aim of the paper was an assessment of the frequency of mutations in the exon 8 of this gene. The specimens from 14 patients operated for glial tumors were investigated by polymerase chain reaction-assisted--single strand conformation polymorphism (PCR SSCP). We found aberrant bands in 64.3% of specimens. The percentage of mutations was similar in patients with benign and malignant tumours. There was no correlation between the alteration of the gene and intensity of necrosis in histological examination in patients with glioblastoma. Changes in activity of the p53 gene were more frequent in younger patients and in males when compared to women. PMID- 11935678 TI - [Expression of bcl-2 gene in glia-derived tumors of the brain]. AB - Bcl-2 gene is an important factor in regulation of apoptosis as an inhibitor of this process. Its increased expression is observed in numerous neoplasms, what points out on importance of this gene in the process of oncogenesis. The aim of this paper was to assess the expression of bcl-2 gene in the cells of the glia derived tumours of the brain. The method of Chomczynski and Sacchi with guanidine tiocyanate was applied to isolate a whole cellular RNA. Expression of bcl-2 gene was measured with a reverse transcription method by a synthesis of cDNA and amplification of gene fragment with specific oligonucleotides in polymerase chain reaction (RT-PCR). The expression of bcl-2 gene was found in cells of all examined glial tumours at the level of transcript. We carried out the discussion of the results and an attempt to explain the importance of bcl-2 gene expression of these neoplasms. PMID- 11935679 TI - [Aneurysms causing symptoms of brain tumors treated with endovascular method in MDS system]. AB - Basing on own material 25 patients with symptoms of intracranial structures pressed by aneurysm sack have been presented. The patients were treated with endovascular method in MDS system at the Department of Neurosurgery, Copernicus Hospital in Lodz. The aneurysms were diagnosed basing on neurological symptoms- cranial nerves damage, focal brain lesion and headaches. The aneurysms surgically difficult and were qualified for endovascular treatment. Ninety two per cent of aneurysms were big or giant; 92% with a wide neck. Total or almost total aneurysm closure was obtained in 59% of cases. The patients were observed 6 to 71 months. Follow up of 11 patients (50% observed group) revealed complete recovery in 8, neurological improvement in 3 cases. PMID- 11935680 TI - [Cerebral arteriovenous malformation complicated by pulmonary embolism]. AB - Endovascular treatment of cerebral arteriovenous malformation (AVM) lies in selective catheterisation through feeder and AVM nidus. Sticking mixture is applied after proper verification of microcatheter location. Malformation embolisation is normal when the vessel net is of small diameter. In the presented case embolisation material penetrated cerebral vein system and then lungs, where numerous emboli developed. X-ray where confirmed pulmonary embolism resulted in severe respiratory insufficiency. Treatment of this rare complication as well as the cause of its occurrence have been presented. Prevention of such complications has been considered. PMID- 11935681 TI - [Stereotactic biopsy and brachytherapy in the diagnostics and treatment of brain tumors--preliminary report]. AB - Promising results have been obtained using brachytherapy in the treatment of brain tumors. Between November 99 and August 2000, 28 patients with brain tumors (15 newly diagnosed gliomas, 11 recurrent gliomas, 2 metastases) underwent implantation of temporary iridium 192 sources with stereotactic technique. This group received external beam radiation therapy (45 Gy) following implantation. Patients were followed-up with CT scans every 3 months. Serious complications occurred in two patients (postradiation brain oedema). Median survival has not been assessed due to short follow-up period and small number of patients. Further clinical assessment is required especially long-term follow-up. Brachytherapy appears to be a useful technique for the treatment of selected brain tumors. PMID- 11935682 TI - [Horner's syndrome induced by internal carotid artery aneurysm--diagnostics and therapy]. AB - Horner's syndrome is most frequently observed in the course of inflammatory, neoplastic and traumatic processes in the area of medulla oblongata, Gasserian ganglion and the lower part of brachial plexus. Horner's syndrome resulting from internal carotid artery aneurysm haemorrhage is very rare. A case of rapid appearance of Horner's syndrome initially treated as contrlateral exopthalmos has been presented. After several weeks headaches and neck pain accompanied Horner's syndrome on the same side. The CT-scan and cerebral angiography were normal. Repeated cerebral angiography with carotid arteries exhibition revealed an aneurysm on the extracranial part of the internal carotid artery. The aneurysm location was considered to be surgically difficult and thus endovascular treatment was applied (coiling). The regression of Horner's syndrome was observed 6 months after the treatment. PMID- 11935683 TI - [Post-traumatic pulsating exophthalmus coexisting with congenital carotid cavernous fistula]. AB - Pulsating exophthalmus develops most frequently in the course of penetrating or bitemporal head trauma. Idiopathic form is even more rare. Orbit venous system over-loading with pathological arterial blood infiltration is the cause of pulsating exophthalmus. A patient with congenital carotid-cavernous fistula (right eyeball enlargement and noise), suffering from head trauma after road accident has been presented. Carotid-cavernous fistula was diagnosed and than treated casually by endovascular closure of the infiltration between internal carotid artery and cavernous sinus. After 5-month treatment regression of the noise and exophthalmus was observed. PMID- 11935684 TI - [The application and evaluation of TachoComb in repair of lacerations of dural sac]. AB - The opening of dural sac (deliberate or accidental) during operation of the spine should be watertightly closed because of the risk of cerebro-spinal fluid leakage and infectious complications. Suturing the dural sac is sometimes difficult and time-consuming in cases of dural sac lacerations in a narrow confined space or in the proximity of radices. This occurs particularly in the case of reoperation in presence of scar involving the dural sac, that increases the risk of laceration. The aim of this paper is to asses the usefulness of a new sealing and haemostatic material--TachoComb in a watertight closure of the dural sac during spinal operation. We compared the frequency of complications related to closing of the dural sac in patients operated before and after introduction of TachoComb. The results indicate that the rate of liquorrhoea and infections is lower in the postoperative period with addition of TachoComb. Addition of TachoComb allows to shorten the time of operation, while its application is relatively easy. PMID- 11935685 TI - [Qualification of patients with CPA tumors to translabyrinthine approach based on own experiences]. AB - On the basis of the presented clinical material, the authors present their own observations concerning qualification of the patients with cerebello-pontine angle tumors for surgery utilizing the translabyrinthine approach. The clinical material includes 15 cases of stage II, III and IV cerebello-pontine angle tumors removed through translabyrinthine approach. In view of the fact that there are different opinions concerning the selection of appropriate surgical approach and of a conviction that translabyrinthine approach should be used in case of stage II, relatively small tumors, and that larger, stage III and IV ones should be operated on using the retromastoid approach, we are presenting our views concerning the qualification of patients. In our opinion, the translabyrinthine approach can be used for larger tumors--also for stage IV ones. The translabyrinthine approach was considered only in the cases with total hearing loss and not useful hearing. Of course, in the cases when the tumor penetrates into the middle cranial fossa or into the foramen magnum, the approach may prove rather difficult. As it has been known, the dimensions of the access route are determined by three main anatomical structures: altitude of the bulb of the internal jugular vein, location of the sigmoid sinus and the diameter of the internal auditory meatus. The fundamental advantages of the approach include: no cerebellum retraction required in the case of classic retromastoid approach, easy anatomical and neurophysiological identification of the course of the facial nerve and maintaining its continuity. An approach involving damage of internal ear structures can be performed only if the patient is deaf. PMID- 11935686 TI - [Indication to translabyrinthine approach to remove of the cerebello-pontine angle tumors--anatomical aspects]. AB - Indication to translabyrinthine approach to removing of the cerebello-pontine angle tumors--anatomical aspects. Translabyrinthine approach was used in surgical procedures performed for stage II, III and IV tumors of the cerebello-pontine angle of various sizes in 15 cases. In order to ensure a correct course of the procedure and successful removal of the tumor, translabyrinthine approach should be carefully planned on the basis of radiological criteria (CT, NMR). Anatomical limitations of the approach are connected with the localization of the sigmoid sinus, the bulb of the internal jugular vein, pneumatization of the mastoid process and the location of the superior petrosal sinus. In approach planning it should be considered whether the bulb of the internal jugular vein is not located high in relation to the internal auditory meatus, what is the meatus diameter (IAC), location of the sigmoid sinus--its shift to the front is a serious limitation for the approach. The pneumatization state of the mastoid process is also important because of the duration of petrosectomy. Taking into consideration the aforementioned conditions allows to plan precisely and perform the approach appropriate for the tumor size. PMID- 11935687 TI - [Neuroprotection in brain injuries]. PMID- 11935688 TI - [Magnetic resonance proton spectroscopy in neurooncology--preliminary report]. AB - MR proton spectroscopy (1H MRS) has been widely applied in characterisation and differentiation of brain tumors, staging, recurrence of pathologic process, post radiotherapy changes and other lesions mimicking neoplasm like abscesses. Despite of many studies performed over last 3 years in many countries spectroscopic pattern (phenotype) of brain tumors is still not well estimated and the role of each metabolite as an indicator of histopathologic grade and type of the tumor is still unknown. MATERIALS AND METHODS: In vivo 1H MR spectroscopy was prospectively performed in 36 patients with intracranial tumors (low grade gliomas, high grade gliomas, meningiomas and abscesses). Examinations was performed with 1.5 T system, using voxels of 8 cc. RESULTS: In high grade gliomas level of NAA was decreased, and consequently low ratios of NAA to other metabolites were obtained; lactate and choline peaks were markedly increased. In patients with meningiomas signal of NAA was reduced, while in abscesses group peaks of acetate and succinate were observed. Authors analysed ratios of metabolites in above-mentioned tumors. The method is especially useful in differential diagnosis in neoplastic and nonneoplastic cases. CONCLUSIONS: Our study confirms diagnostic value of 1H MRS in doubtful cases of intracranial tumors. PMID- 11935689 TI - [Retinopathy of prematurity--frequency and risk factors]. AB - Retinopathy of prematurity (ROP) is a significant problem in very low birth weight (VLBW) newborns, patients of the neonatal intensive care units (NICU). The aim of the present investigation is to report the results of our ROP-screening program: 1. Current frequency of ROP and the need for retinal surgery; 2. Main risk factors and the prophylactic possibilities for ROP. STUDY DESIGN: This is a retrospective study for 3 years period: 08.'98-07.01. 188 survived premature newborn patients of NICU, "Majchin dom", Sofia, met the criteria for ROP screening. The first eye exam was performed at 3 weeks of age, followed by exams every 2 weeks until complete vascularisation of the retina. RESULTS: The incidence of ROP any stage was 11.2% (21 of 188) and 4.2% (8) for stage > or = 3. The mean birth weight of the ROP (+) group was 982 g, the mean gestational age (g.a.)--27.9 g.w., comparing with 1258 g and 30.9 g.w. respectively of the ROP(-) group (p < 0.05). ROP stage > or = 3 was diagnosed as a rule among the newborns with birth weight < 1000 g 14.6% (7 of 48) and g.a. < 28 g.w. (6 of 41). Seven children have undergone successful retinal surgery. The children of ROP(+) group needed significantly longer mechanical ventilation and supplemental O2, they developed more often bronchopulmonary dysplasia and severe grade III-IV IVH, the percent of neonatal sepsis and the need of blood transfusions in the first month was greater too. CONCLUSION: The frequency and severity of ROP in our study is relatively low and that depends on the strict monitoring of the intensive cares. Main risk factors are extremely low birth weight and gestational age; severe RDS with longer duration of mechanical ventilation and supplemental oxygen. Grade III IV IVH and neonatal sepsis are often a parallel finding with severe ROP. Other risk factors are the greater number of blood transfusions and the developing of chronic lung disease. PMID- 11935690 TI - [von Recklinghausen's disease and pregnancy]. AB - Neurofibrosis or the Von Recklinghausen disease constitutes a genetically transmitted disease that appears in many parts of the body as well as many symptoms. It is inherited from the autosomatic dominant type (gene NFI on chromosome 17) and is characterized by brownish blemishes of the skin (cafe au lait), multiple neurofibromas (small dermal or subcutaneous lumps), symptoms of the peripheral and central nervous systems as well as disorders of the eyes, skeletal system, gastric system and vascular system (aneurysms of the aorta, carotid and renal arteries). We describe the case study of a 25 year old female, second pregnancy with known history of the above mentioned disease. She delivered on the 39th week of pregnancy (male neonate weighing 3000 gr.), with no problems. On the 22nd week of her pregnancy she presented with a large distension of the anterior surface of the neck in the area of the thyroid that gradually increased in size. Differential Diagnosis includes nodular goiter and neurofibroma. One month postpartum the morphoma was surgically. Neurofibrosis during pregnancy can cause many complications such as IUGR, hypertensive disease, spontaneous abortions, oligohydraminios and neurofibroma development. This case study is being discussed in international bibliography. PMID- 11935691 TI - [Dysfunctional uterine bleeding in adolescence]. AB - Dysfunctional Uterine Bleeding (DUB) is a frequent gynecological problem during adolescence. In about 95% of cases it is caused by the late maturation of the hypothalamic-pituitary-ovarian axis (HPO), leading to anovulatory cycles. These adolescents lack the E2 positive feedback on LH. Thus, the continuous production of estrogen with endometrial stimulation is the basic cause of DUB. The initial step in the evaluation of the disease includes detailed clinical history, followed by complete physical examination. Laboratory tests should include coagulation profile, complete blood count with platelet evaluation, and sometimes a serum pregnancy test. The treatment of DUB is related to the severity of symptomatology with the objective of stopping bleeding and preventing recurrences. The importance of continued follow-up in DUB cases should be underlined, until stabilization of ovulatory menstrual cycles. PMID- 11935692 TI - [Treatment of Chlamydia trachomatis with azatril]. AB - The authors discuss the treatment with Azatril for Chlamydia trachomatis infection in patients with tubar sterility. The study includes 85 women, treated for three months. The authors achieved 57.6% negativation of the results that gives a reason to propose the use of Azatril (Balkanpharma) in the treatment of Chlamydial infection. PMID- 11935693 TI - [Treatment of chlamydial cervicitis with azatril]. PMID- 11935694 TI - [Borderline ovarian tumors]. AB - Forty three patients with borderline ovarian tumours were examined from 1987 till 2000. 28 patients (65.1%) were serous tumours and 15 patients (35%) were mucinous. The median age was 42.2 +/- 15.4 years. The follow-up period was 5.4 +/ 3.2 years. Preoperatively Ca125 were 52.5 +/- 50.3 and of Ca19.9-42.4 +/- 46.8 IU. 83% of the patients were in stage I and 17% in stage III. There was no significant difference in the tumour volumes. Ca125 were significantly increased in the serous tumours and Ca19.9 were increased in the mucinous tumours. All the patients were followed up and are in good health or remission with exception of 2 who developed recurrence. PMID- 11935695 TI - [Selection criteria for breast conservation in patients with early breast carcinoma]. AB - During the past two decades, breast-conserving therapy (excision of the tumor and axillary lymphadenectomy followed by irradiation) for early stage breast carcinoma has become firmly established as an equivalent treatment approach to mastectomy. The purpose of this review as to examine the risk factors for local recurrence after breast-conserving therapy. Better mammographic evaluation, better margin assessment, recognition of an extensive intraductal component and the use of adjuvant systemic therapy has improved the logo-regional control. PMID- 11935696 TI - [Structure and function of epidermal growth factor receptors and their significance as a prognostic marker in ovarian carcinoma]. PMID- 11935698 TI - [DNA multiparametric analysis by flow cytometry of cancer of the uterine cervix]. PMID- 11935697 TI - [Characteristics of neonates whose mothers have congenital and acquired heart disease]. AB - It was made a study of the body dimensions, Apgar evaluation, and frequency of innate body defects in 141 new-borns by mothers with innate and acquired heart valvular defects. It was found that the average values of body weight, height, and the evaluation with Apgar of the new-borns by mothers with heart defects do not deviate from the average values of the same indices of new-borns by healthy mothers. Despite that 90% of the young mothers with heart defects are with light to moderate degree of the heart defect, and with I or II functionality class, the percent of their new-borns born prematurely is big--31.9, compared to 6.6 for new borns by healthy mothers. In 2.8% of the new-borns by mothers with heart defects, hypotrophy is observed, and in 3.5%--clinical criteria for immaturity. Perinatal death rate is 0.7% and zero for the control group of new-borns--ones born by healthy women. Six cases (6.7%) of new-borns by mothers with heart defects had innate body defects, 4 (4.5%) of which were innate heart defects. The frequency and the kind of the congenital defects in new-borns by mothers with heart defects is discussed. PMID- 11935699 TI - [A case of placenta percreta with spontaneous rupture of the uterus in the second trimester of pregnancy]. AB - The authors present a case with placenta percreta demonstrated with massive intraabdominal haemorrhage in the second trimester of pregnancy. Supravaginal hysterectomy has been performed. The possibilities of early diagnosis of this life threatening condition and the current knowledge about the problem are discussed. PMID- 11935700 TI - [Adamantiades-Behcet's syndrome and pregnancy: a case report]. AB - Is introducing a case study on a 20 year old female first time pregnant who presents with rare case of Adamantiadis-Bechet syndrome. These symptoms first appeared at the age of 9. The patient refers that from this age she has suffered from recurrent relapses of ulcerations of the mouth and genital organs, vescicular exanthema of the upper and lower extremities and severe headaches that would not subside with analgetic therapy. The patient upon the first trimester presented with a relapse of the disease and significant improvement during the remainder of the pregnancy. She delivered on the 39th week of her pregnancy a neonate male, alive and weighing 3640 gr. The Apgar--score 9 and 10 the 1st and 5th minute. Mother and neonate were discharged from the clinic on the 3rd postpartum day in good health. This case is also being discussed in international bibliography. Adamantiadis-Behcet syndrome constitutes systemic angiitis of unknown etiology that is characterized mainly by recurrent ulcers of the mouth and genital organs, serum negative arthritis, central nervous system disorders as well as ulcerations of the epidermis, nodular erythema, thrombophlebitis and vision disorders. It usually affects people in early adulthood, more often those in their 30s. Geographically it is seen more often in Japan, Korea, Turkey, Israel and other Mediterranean countries. Familial prevalence is also common. PMID- 11935701 TI - [Analysis of indications for a repeat Caesarean section]. AB - The aim of the study is to establish the indications for a Resectio Caesarean (RS) and to compare them with those of the first Caesarean section (CS). The Study is retrospective and covers the period from 1.1.2000 to 31.XII.2000. For this period were accomplished 3646 deliveries, 894 (24.52%) of them with S. C. The RS were 182 (20.04%) of all CS. The number and the rate of each indication is calculate on the base of the sum of the first and the second indication from all CS. The first two indications reflect well the real reason for each CS. The results show that the average age of women with RS is grater 2 years and 7 months than those with first CS. The RS are elective in 80.7% of cases and is done at 38.4 +/- 1.4 w.g. The average weight of the newborns from RS is non significantly less with 31 g than this from first CS. The indication "status post CS" is not well defined as it is used in 98.3% of RS. The indication is accepted mostly as a relative one because is added in cases with enough reasonable other obstetric indications. The analysis of the indications of RS shows That 60% of them are justifiable 40% are undertaken on the bases of unconvincing complex indication, but which don't differ from those at first CS. PMID- 11935702 TI - Walk-in clinics and time management. Fresh insights as family practices adapt. PMID- 11935703 TI - Primary care reform. View from Australia. PMID- 11935705 TI - Exercise and children with asthma. PMID- 11935704 TI - Preference for the no-stirrup method. PMID- 11935706 TI - Nicotine patches and pregnancy. PMID- 11935707 TI - Mental health care in aboriginals is neglected. PMID- 11935708 TI - Cystic fibrosis during pregnancy. AB - QUESTION: I have a 23-year-old patient with cystic fibrosis (CF) who recently married and has come to see me regarding preconception counseling and prospects of pregnancy. According to my sources, patients with CF can have successful pregnancies. How should I advise my patient? ANSWER: Cystic fibrosis in all degrees of severity is not a strict contraindication for pregnancy, especially if disease is mild; pregnancy by itself does not appear to adversely affect patients with CF. Pregnancy could proceed normally in women with normal lung function, but could adversely affect mild and moderate lung disease due to CF and should be avoided when patients have pulmonary hypertension or cor pulmonale, and when reduced lung function is predicted. Ideally, all pregnancies should be planned with prior counseling and be monitored by dedicated CF teams. PMID- 11935709 TI - Ophthaproblem. Vitamin supplementation. PMID- 11935710 TI - Reducing anterior shoulder dislocation. Easy is good. PMID- 11935711 TI - Just the berries. Ecstasy. PMID- 11935712 TI - Taking NSAIDs during pregnancy. Is it safe? PMID- 11935713 TI - Analysis of serious adverse events. Lipid-lowering therapy revisited. PMID- 11935714 TI - Managing diabetes in childhood and adolescence. AB - OBJECTIVE: To describe management of children's and adolescents' diabetes outlining standards of care compatible with current clinical practice. QUALITY OF EVIDENCE: MEDLINE was searched using specified MeSH headings. Bibliographies of selected articles were used to find additional pertinent articles. Preference was given to randomized controlled trials, clinical practice guidelines, consensus statements, and task force recommendations. We also cite reviews of current practice regarding pediatric diabetes. MAIN MESSAGE: Managing children with diabetes presents a difficult challenge to parents and their advisors. Achieving good diabetic control is impossible unless parents are properly instructed in practical management of the disease. Children with diabetes should be managed quite differently from adults in several respects. Avoiding hypoglycemia is most important, particularly for preschool children. Higher target blood glucose levels than would be accepted for adults are both justifiable and necessary for preschool children. Controlling children's diabetes depends as much on personal factors and family adjustment as it does on insulin, food plans, and exercise. CONCLUSION: Diabetes mellitus is difficult to manage at any age. Managing children's diabetes successfully requires continuous education and encouragement of parents and children. Pediatric diabetes care teams and family physicians play a vital role in encouraging children to control their disease while participating fully in normal childhood activities. PMID- 11935715 TI - [Disorders related to traumatic events. Screening and treatment]. AB - OBJECTIVE: To educate family physicians about screening, diagnosis, and treatment of psychological disorders related to traumatic events. QUALITY OF EVIDENCE: PsycLIT, PsychINFO, PILOTS, and MEDLINE databases were searched from January 1985 to December 2000 using the terms "acute stress disorder," "posttraumatic stress disorder," "traumatic stress," "psychotherapy," "psychosocial treatment," "treatment," and "pharmacotherapy." Recommendations concerning treatment of acute stress disorder (ASD) and posttraumatic stress disorder (PTSD) are based on evidence from trials of the highest quality. Conclusions about assessment and diagnosis are based on the most recent epidemiologic studies, consensus, and expert opinion. MAIN MESSAGE: Very often, ASD and PTSD are underdiagnosed and undertreated. Family physicians are likely to see patients suffering from these disorders. Early screening in primary care is a function of active listening; warm, safe patient-physician relationships; and careful examination of difficulties related to traumatic events. Ideally, patients with either ASD or PTSD should be referred to a specialist. If a specialist is unavailable, family physicians can offer support and prescribe medication to address patients' symptoms. CONCLUSION: Family physicians can help identify and treat patients presenting with disorders related to traumatic events. PMID- 11935717 TI - Walk-in clinics in Ontario. An atmosphere of tension. AB - OBJECTIVE: To explore family practice (FP), emergency department (ED), and walk in clinic (WIC) physicians' perceptions and experiences regarding the effect of walk-in clinics on Ontario's health care system. DESIGN: Qualitative method of focus groups. SETTING: Hamilton, London, and Toronto, Ont. PARTICIPANTS: Sixty three physicians participated in nine focus groups, each with four to nine participants. Family physicians, ED physicians, and WIC physicians attended separate focus groups. METHOD: Nine focus groups were conducted in three cities in Ontario. Physicians' opinions, perceptions, and experiences regarding the role and effect of WICs on Ontario's health care system were explored. Focus groups were audiotaped and comments transcribed verbatim. The qualitative data analysis program NUD*IST was used to organize the data during sequential thematic analysis. MAIN FINDINGS: Participants identified two key factors contributing to the evolution of WICs: patients' expectations for convenient health care and the perceived limited availability of family physicians. Participants thought these two related factors resulted in a gap in primary care services that WICs had filled. Throughout discussions, an atmosphere of tension permeated the focus groups. Tension seemed to arise from issues of duplication, competition, standards of practice and quality of care in WICs, the effect of environmental and personal factors on physicians' practice, and the practice philosophy adopted by WIC physicians. CONCLUSION: Both FP and ED participants acknowledged their contribution to the gap in primary care services. They appeared to attribute current problems in health care delivery to the perceived deficiencies of WICs. The outcome was a marked tension among participants. PMID- 11935716 TI - Who provides walk-in services? Survey of primary care practice in Ontario. AB - OBJECTIVE: To compare walk-in clinics with other primary care settings on characteristics associated with best practices in primary care. DESIGN: A mailed self-administered questionnaire asked about organizational and clinical characteristics of primary care practices located in major urban and suburban areas in Ontario. SETTING: Four types of fee-for-service group practices: walk-in and urgent-care clinics (WICs), mixed practices (MPs), after-hours clinics (AHCs), and group family practices (GFPs). PARTICIPANTS: A physician or a staff member involved in practice administration. MAIN OUTCOME MEASURES: The four practice types were compared on organizational characteristics and measures of access, continuing care, comprehensiveness, coordination, and mechanisms for monitoring quality of care. RESULTS: Walk-in clinics, MPs, and AHCs were open more hours during evenings and weekends and were more likely to see patients without appointments; GFPs were more likely to have on-call arrangements. Group family practices saw a larger proportion of patients for whom they provided ongoing care; WICs and MPs reported that more than 60% of their visits were with "regular" patients. Walk-in clinics were less likely to provide preventive services and psychological counseling than were GFPs and MPs. A few WICs, MPs, and GFPs had procedures to support coordination of care or to monitor quality of care. CONCLUSION: Although WICs, MPs, and AHCs provided walk-in services to Ontario patients, WICs and MPs also provided a substantial amount of ongoing care and preventive services. Independent AHCs appeared to most closely fit the "walk in clinic" stereotype. PMID- 11935718 TI - Medical abortion and family physicians. Survey of residents and practitioners in two Ontario settings. AB - OBJECTIVE: To determine the knowledge, attitudes, and interest in providing medical abortion reported by family physicians and residents in rural and urban settings. DESIGN: A self-administered mailed survey using the modified Dillman method. SETTING: Hamilton and Thunder Bay County in Ontario. PARTICIPANTS: Family medicine residents (n = 93) and physicians (n = 234) in predominantly urban (Hamilton) and rural (Thunder Bay) settings. All faculty family physicians at McMaster University practising general family medicine and all family physicians in Thunder Bay County were surveyed. MAIN OUTCOME MEASURES: Knowledge of, attitudes toward, and interest in providing medical abortion. RESULTS: Overall response rate to the survey was 62.7% (n = 327); 74.2% (69/93) of residents responded; 58.1% (136/234) of physicians responded. Physicians and residents rated their knowledge about medical abortion as poor, but most were interested in receiving more information and training in this area. Many (83.1%, 157/189) reported that medical abortion was an acceptable procedure for family physicians to perform, and 52.0% (64/123) of the physicians would consider providing medical abortions for their patients. Residents training in the more rural Thunder Bay program were less likely to support first-trimester abortions for both medical and nonmedical reasons than those training in Hamilton (P < .05). Male respondents were significantly less supportive of abortion for nonmedical reasons and were less likely to consider providing medical abortions for their patients (P < .05). CONCLUSION: Most family physicians and residents showed interest in receiving more information about and training in medical abortion. PMID- 11935719 TI - Caring for our own families. PMID- 11935720 TI - Hypothesis: the research page. Participatory methods to facilitate research. PMID- 11935721 TI - Transferring medical records: improving the exchange. PMID- 11935722 TI - The accuracy and reliability of the tympanic membrane thermometer. A literature review. PMID- 11935723 TI - Cwilted. PMID- 11935724 TI - Theophylline overdose: clinical features and management. PMID- 11935725 TI - A hybrid emergency care model. PMID- 11935726 TI - Devising a patient group direction. PMID- 11935727 TI - Returning to practice--a lecturer's experience. PMID- 11935728 TI - Managing emergency admissions. PMID- 11935729 TI - Dynamics of future working. PMID- 11935730 TI - Button battery exposure: clinical features and management. PMID- 11935731 TI - How much is enough? Emergency oxygen therapy with COPD. PMID- 11935732 TI - Physiological responses and management of hypothermia. PMID- 11935733 TI - Thrombolytic therapy: an A&E perspective. PMID- 11935734 TI - The move in the United States towards generic emergency workers. PMID- 11935736 TI - Differing attitudes to care. PMID- 11935737 TI - Nurse consultants in A&E. PMID- 11935735 TI - Suitable cases for teleconsultation by ENPs. PMID- 11935738 TI - Silent conflict. Helping the survivor of sexual violence. PMID- 11935739 TI - Antimalarial drug overdose: clinical features and management. PMID- 11935740 TI - Non accidental injury vs staphylococcal scalded skin syndrome. A case study. PMID- 11935741 TI - Prompts to request telemedical consultations in clinical protocols. PMID- 11935742 TI - Penetrating trauma. A case of the unexpected. PMID- 11935743 TI - Casualty rethink after rail crash disaster. PMID- 11935744 TI - Emergency nursing on the Everest base camp trek. PMID- 11935745 TI - Degrees of choice. PMID- 11935746 TI - Extending the ENP role with ENP review clinics. A preliminary study. PMID- 11935747 TI - Digoxin overdose: clinical features and management. PMID- 11935748 TI - A&E liaison mental health nursing: a working reality. PMID- 11935749 TI - Emergency care nurses win new prescribing freedom. PMID- 11935750 TI - Great expectations. A survey of the expectations of self-referred patients and their accompanying relatives and friends of an A&E service: Part one. PMID- 11935751 TI - Cot death guidelines published for A&E staff. PMID- 11935752 TI - Seeing the bigger picture. PMID- 11935753 TI - Down under: emergency nursing in Australia. PMID- 11935754 TI - USA under attack. Coping with the aftermath. PMID- 11935756 TI - Self-care. Will 'expert patients' undercut nurses? PMID- 11935755 TI - You can run, but you can't hide. PMID- 11935757 TI - Elderly care. Bordering on the ridiculous. PMID- 11935758 TI - Should nurses in uniform be banned from smoking? PMID- 11935764 TI - Cop a load of this. PMID- 11935765 TI - The more things change.... PMID- 11935767 TI - A focus on innovation to boost national care standards. PMID- 11935766 TI - Home truths. PMID- 11935768 TI - Staff development through appraisal. PMID- 11935769 TI - Mental illness and substance abuse. PMID- 11935770 TI - How to create a support network for research nurses. PMID- 11935771 TI - Developing the nursing role in major trauma. PMID- 11935773 TI - Changing behaviour through relocation. PMID- 11935777 TI - Libyan HIV trial is postponed. PMID- 11935774 TI - Caring for ventilated children at home. PMID- 11935781 TI - Providing dental services to children enrolled in the Hawk-I Program. PMID- 11935782 TI - HHS clarifies HIPAA privacy guidelines. PMID- 11935783 TI - Lasers use OK receives mixed reaction. IDA awaiting more information. PMID- 11935784 TI - HMO's call for binding reforms on managed care industry--"PARCA" targeted. PMID- 11935785 TI - "Principles of professional ethics". PMID- 11935786 TI - New rules will affect transmission of electronic information. PMID- 11935787 TI - 1,395 private practice dentists in state of Iowa. PMID- 11935788 TI - Iowa sports dentistry. PMID- 11935789 TI - "What impact will the separation of Delta and BC/BS have on customers and providers?". PMID- 11935790 TI - So you don't think we have a public relations problem! PMID- 11935791 TI - A bright future for dentistry and dental education. PMID- 11935792 TI - Health task force issues recommendations. Elimination of mandatory CEU's possible. PMID- 11935793 TI - Online continuing education. Know what you're getting. PMID- 11935794 TI - Congress acts to regulate needles--state to follow. PMID- 11935795 TI - New rules & regulations on dental assistants. PMID- 11935796 TI - OSHA issues new ergonomic rules. PMID- 11935797 TI - Nitrous requirements revisited. PMID- 11935798 TI - Language ability affects access--HHS issues policy guidance. PMID- 11935799 TI - Above and beyond the call of duty. PMID- 11935800 TI - Results of the 1999 Iowa oral health survey. PMID- 11935801 TI - The radiograph guidelines for new pediatric patients. How well are we doing? PMID- 11935802 TI - Are DMSO's in Iowa's future? PMID- 11935803 TI - "The greater good". PMID- 11935804 TI - SPRC could spell changes in dental offices. PMID- 11935805 TI - State finalizes rules on x-ray testing. PMID- 11935806 TI - Local anesthesia rules finalized. PMID- 11935807 TI - Dental practitioner demographics preliminary report 1998. PMID- 11935808 TI - Intraoral/perioral piercing. Implications for the dental professional. PMID- 11935809 TI - Crystal methamphetamine and the dental patient. PMID- 11935810 TI - The Iowa Dental Medicaid program. Legislature acts to improve access & reimbursement. PMID- 11935811 TI - Does soda pop cause cavities? PMID- 11935812 TI - Dental assisting legislation becomes law. PMID- 11935813 TI - Dental waterline safety questioned on national TV. PMID- 11935814 TI - ADA recognizes new dental specialty. PMID- 11935815 TI - Where have we been this last century? PMID- 11935816 TI - Does big equal bad? PMID- 11935817 TI - Impaired practitioner program. PMID- 11935818 TI - Hyperbaric oxygen therapy and osteoradionecrosis. AB - Osteoradionecrosis of the mandible is a potentially devastating complication of head and neck radiation. Radiation causes progressive vascular occlusion with tissue hypoxia, tissue death, and failure of healing. Hyperbaric oxygen therapy is an accepted treatment for osteoradionecrosis. This article reviews the history of hyperbaric oxygen therapy, its physiologic mechanisms, its use in the management of osteoradionecrosis, and its complications and contraindications. PMID- 11935819 TI - Acute poisoning: bleaches, disinfectants and detergents. PMID- 11935820 TI - Implementation of an education & development strategy & audit tool in the Emergency Department. PMID- 11935821 TI - Patient satisfaction with Emergency Nurse Practitioners. PMID- 11935822 TI - What does the triage nurse do? PMID- 11935823 TI - Working smarter, not harder. Improving emergency department nurse productivity. PMID- 11935824 TI - What about us? The psychological implications of dealing with trauma following the Omagh bombing. PMID- 11935825 TI - New AIDS vaccine developed for Africa. PMID- 11935831 TI - Dance with them, marry them, but don't go into business with them. PMID- 11935832 TI - The patient-centered plan of care. PMID- 11935833 TI - CareScience offers products for complex clinical data management. PMID- 11935834 TI - Some thoughts on the art of healing. PMID- 11935835 TI - How to search for and find useful health care information on-line. PMID- 11935836 TI - Outcomes in high risk pregnancy and neonatal case management. PMID- 11935837 TI - Collaboration: an innovative education/business partnership. AB - Healthy People 2010 initiatives encourage the collaborative partnership goals described in this article. The partnership developed between a UB School of Nursing faculty member, her students, and several case managers at Health-Now Now has been a richly rewarding experience. The opportunities afforded by this partnership have expanded experiences for faculty, students, and case managers in a managed care environment. This partnership has enabled participants to initiate practical, cost-effective methods for improving community-based services. These partnerships should yield significant changes in health behaviors and health outcomes among the American public. Valanis states it most succinctly: "The nurse of the 21st century must innovate, coordinate, and monitor services for populations within the health care system in which she works and interacts with crucial services outside the system. Her or his patient is not only an individual or even the family but the entire community." PMID- 11935839 TI - The rebirth of case management. PMID- 11935838 TI - Minimizing the impact of chronic obstructive pulmonary disease. AB - COPD, a common disease with potentially devastating consequences, is both preventable and treatable. The key to successful disease management is differential diagnosis, and the most important intervention is smoking cessation. Medications can be used to reduce symptoms, vaccines to prevent life-threatening exacerbations, and oxygen to treat hypoxemia. Diet and exercise also may help improve outcomes. PMID- 11935840 TI - Staff retention in a tight labor market. PMID- 11935841 TI - Hospital: a place of safety? PMID- 11935843 TI - Gays lead fight for an equitable life. PMID- 11935842 TI - Child health. Out of sight, out of mind. PMID- 11935844 TI - Elderly care. No benefit from ill-gotten gains. PMID- 11935846 TI - Should cesareans be performed only on the basis of medical need? PMID- 11935845 TI - Hail the unsung heroes. PMID- 11935847 TI - Who's responsible for students' mistakes? PMID- 11935848 TI - Taking note. PMID- 11935849 TI - Working on the home front. PMID- 11935850 TI - The heart of the community. PMID- 11935851 TI - Privatisation: the final solution. PMID- 11935852 TI - Auditing PICC line management. PMID- 11935853 TI - Getting to grips with writing reports. PMID- 11935854 TI - Communicating effectively with psychotic patients. PMID- 11935855 TI - Risk assessment in theatres. PMID- 11935856 TI - Several women students were diagnosed with chlamydial infections last term so we are keen to promote awareness of this condition. PMID- 11935857 TI - Managing asthma in adolescence. PMID- 11935858 TI - Latex allergy awareness among hospital staff. PMID- 11935860 TI - Lymphatic filariasis. PMID- 11935859 TI - MRSA: false economy may lead to dangerous practices. PMID- 11935862 TI - HIV infection: five face death if guilty. PMID- 11935861 TI - Administering antibiotic therapy at home. PMID- 11935863 TI - Stop smoking campaign. No butts revisited. PMID- 11935864 TI - Should the NHS send patients abroad to reduce waiting lists? PMID- 11935865 TI - Anybody can make a difference, darling. PMID- 11935866 TI - Living history. PMID- 11935867 TI - Short-changed? PMID- 11935868 TI - A matter of life and death. PMID- 11935870 TI - Thinking your way to successful problem-solving. PMID- 11935869 TI - A way forward for nurse recruitment. PMID- 11935871 TI - Engaging nurses in clinical research. PMID- 11935873 TI - Day in the life. PMID- 11935874 TI - Child friendly. PMID- 11935872 TI - The therapeutic benefits of a women-only environment. PMID- 11935875 TI - Case study: working with adolescents. PMID- 11935878 TI - Study of a nurse practitioner working in a paramedic role. PMID- 11935876 TI - Free nursing care. DoH: nursing not means tested. PMID- 11935877 TI - Violence targets 'unachievable'. PMID- 11935879 TI - The use of Lorazepam in status epilepticus. PMID- 11935880 TI - Children living with domestic violence. PMID- 11935881 TI - Venomous snake bites. PMID- 11935882 TI - UK emergency services on alert. PMID- 11935883 TI - Interagency co-operation in pre-hospital care. PMID- 11935884 TI - Emergency nursing in Sweden. PMID- 11935885 TI - Lessons from the past. PMID- 11935886 TI - Minor injuries/illness nurse treatment service. PMID- 11935888 TI - Coronary heart disease. PMID- 11935887 TI - Antidotes. PMID- 11935889 TI - NHS direct nurse to face UKCC after diabetes death. PMID- 11935890 TI - ENs spell out bioterror procedures. PMID- 11935891 TI - Comparing performance of ENPs and SHOs. PMID- 11935893 TI - Smut fungi (Ustilaginomycetes and Microbotryales, Basidiomycota) in Panama. AB - This is the first publication dedicated to the diversity of smut fungi in Panama based on field work, the study of herbarium specimens, and references taken from literature. It includes smuts parasitizing cultivated and wild plants. The latter are mostly found in rural vegetation. Among the 24 species cited here, 14 species are recorded for the first time for Panama. One of them, Sporisorium ovarium, is observed for the first time in Central America. Entyloma spilanthis is found on the host species Acmella papposa var. macrophylla (Asteraceae) for the first time. Entyloma costaricense and Entyloma ecuadorense are considered synonyms of Entyloma compositarum and Entyloma spilanthis respectively. For the new combination Sponsorium panamensis see note at the end of this publication. Descriptions of the species are complemented by some illustrations, a checklist, and a key. PMID- 11935895 TI - [Six new species and two new records of (Pteridophyta) from Costa Rica ]. AB - Six new species of Pteridophyta from Costa Rica are described: Dryopteris flaccisquama A. Rojas, Hypolepis lellingeri A. Rojas, H. moraniana A. Rojas, Melpomene alan-smithii A. Rojas, Selaginella osaensis A. Rojas and Terpsichore esquiveliana A. Rojas. Blechnum stoloniferum (Mett. ex E. Fourn.) C. Chr. and Trichomanes micayense Hieron. are new records for the country. Only D. flaccisquama and M. alan-smithii are present outside Costa Rica. PMID- 11935894 TI - [Reports in Pteridaceae (Filacales) family in the Neotropics]. AB - Several contributions to the Neotropical pteridophyte flora are made in this paper with the range extensions for four species of Adiantum, two species of Doryopteris and one species of Eriosorus. The species concerned are: A. polyphyllum Willd. for Nicaragua and Costa Rica; A. trichochlaenum Mickel et Beitel for Guatemala and Costa Rica; A. villosissimum Mett. ex Kuhn for Costa Rica; A. wilesianum Hook. for Nicaragua; D. nobilis (T. Moore) C. Chr. for Costa Rica, Colombia and Venezuela; D. redivida Fee for Panama; and E. hirtus (Kunth) Copel. for Costa Rica. The concept of D. pedata (L.) Fee is redefined. PMID- 11935896 TI - [New species, newly used names and new ranges of tree ferns (Filicales: Cyatheaceae) in the Neotropics]. AB - Two new species are described for the Neotropics, four species are newly accepted, and four species and one variety are reported. The new species are: Cyathea grayumii A. Rojas and C. panamensis A. Rojas. Cyathea alfonsiana L. D. Gomez, C. holdridgeana Nisman & L. D. Gomez, C. onusta H. Christ and C. squarrosa (Rosenst.) Domin are recognized; Cnemidaria coclena Stolze, Cyathea andina (H. Karst.) Domin, C. caracasana var. meridensis (H. Karst.) R. M. Tryon, C. macrosora (Baker) Domin and C. pseudonanna (L. D. Gomez) Lellinger are reported from Costa Rica and Panama. PMID- 11935897 TI - [Eight new species and new ranges of Tectariaceae (Felicales) in the Neotropics]. AB - The following species of Tectariaceae (Filicales) from Nicaragua, Costa Rica, Panama, Colombia and Peru are described as new: Ctenitis sotoana A. Rojas, Megalastrum ctenitoides A. Rojas, Tectaria darienensis A. Rojas, T. faberiana A. Rojas, T. longipinnata A. Rojas, T. murilloana A. Rojas, T. pascoensis A. Rojas and T. subdimorpha A. Rojas. The new combination of M. karstenianum (Klotzsch) A. Rojas is proposed. The distributions of C. bullata A. R. Sm., C. microchlaena (Fee) Stolze, C. submarginalis (Langsd. et Fisch.) Ching, T. andina (Baker) C. Chr. and T. sodiroi (Baker) Maxon are enlarged. PMID- 11935898 TI - Local abundance and regional distribution of tree species of forest fragments in Brazil: a test of models. AB - Patterns of local abundance and regional distribution of tree species were analyzed on three spatial scales and compared with those predicted by Hanski's 1982 original core-satellite model, Brown's 1984 niche-based model, and Tokeshi's 1992 model. Data were collected in seven forest fragments dispersed over approximately 7200 km2, remnants of a former continuous forest in the south of Mato Grosso do Sul state, southwestern Brazil. Fifty-six east-west-oriented 10 m yen 150 m plots were randomly demarcated in those fragments from 1994 to 1996. In each plot all trees having DBH > or = 0.15 m were recorded and measured. The smallest scale was that of the fragments themselves and the greatest was that of the entire region. No evidence was found of the bimodality predicted by Hanski's 1982 model on any scale. On all scales, an unimodal pattern was found whose mode was on the left side (satellite mode) of the distribution. This pattern was in best agreement with the predictions of Tokeshi's 1992 model. PMID- 11935899 TI - Biological activity of Ruta chalepensis (Rutaceae) and Sechium pittieri (Cucurbitaceae) extracts on Hypsipyla grandella (Lepidoptera: Pyralidae) larvae. AB - Biological activity of a plant extract (common rue, Ruta chalepensis) and a semi purified fraction (from "tacaco cimarron", Sechium pittieri) on mahogany shootborer larvae (Hypsipyla grandella) was studied. A randomized complete block design, with four replications, was used. H. grandella third instar larvae were exposed for 24 h to Cedrela odorata leaf discs dipped in several treatment dissolutions of each extract (0.1, 0.32, 1.0, 3.20, and 10%); afterwards, each larva was transferred to a flask containing an artificial diet and was allowed to complete its development. Variables measured included food consumption (foliar area eaten in 24 h), mortality, and developmental effects (developmental time for each larval instar and the pupa, and pupal weight). The common rue extract showed a clear antifeedant activity at a concentration as low as 0.32%, whereas the "tacaco cimarron" fraction caused toxicity, especially at the two highest concentrations (3.20 and 10%). PMID- 11935900 TI - Volatile essential oil constituents of Alpinia smithiae (Zingiberaceae). AB - The composition of leaf and rhizome essential oils of Alpinia smithiae that grow wild in the Western Ghats of Kerala (South India) was analysed by gas chromatography. The major components were beta-caryophyllene, sabinene, myrcene and 1,8-cineole in both samples, but variation in the yield of oil as well as the major components between the two plant parts was observed. PMID- 11935901 TI - [Effects of herbivory on ovum number in Loasa speciosa (Loasaceae)]. AB - We examined the direct effect of foliar herbivory on female fitness in the herbaceous plant Loasa speciosa (Loasaceae), measuring ovule number/flower in 46 shoots with different levels of leaf damage. Ovule number/flower was lower in shoots with higher herbivory levels. This result suggests that the known negative impact of herbivory on female plant fitness (i.e. low seed number) can be a direct consequence of the reduction of ovule number. PMID- 11935902 TI - Effect of storage temperatures on growth and survival of Escherichia coli O157: H7 inoculated in foods from a neotropical environment. AB - Escherichia coli O157: H7 has emerged as a new pathogen and is found worldwide. We studied the effect of several storage temperatures on the survival of this bacterium in common foods from a neotropical environment (Costa Rica) because at least seven clinical cases have been reported from the country, and no epidemiological link or probable food association has been described. High (10(6) 10(8) CFU/ml) and low (10(4)-10(6) CFU/ml) populations of E. coli were inoculated (three replications) in ground meat, chopped cabbage, chicken giblets and pasteurized milk and incubated at 0, 6 and 12 degrees C for 24, 48 and 72 h. Vegetables and milk were also stored at 22 degrees C for the same periods. The E. coli O157: H7 enumeration was done according to the methodology described in the Bacteriological Analytical Manual. Populations of E. coli O157: H7 showed either an increasing or decreasing trend, according to temperature, time or food base. Our data indicate that E. coli O157: H7 is capable of surviving and growing in meat, cabbage, milk and chicken giblets; food items commonly consumed by Costa Ricans. PMID- 11935903 TI - Anoxia in a coastal bay: case study of a seasonal event. AB - Bahia Concepcion (Gulf of California) was studied to describe some extremely low dissolved-oxygen values at the bottom of the bay. Surveys included measurements of vertical distribution of temperature, dissolved oxygen, nitrate, nitrite, phosphates and hydrogen sulfide. Thermal stratification from early spring to autumn and a well-mixed water column during the winter were present. Dissolved oxygen concentration was homogeneous in the water column from winter to spring (5 7 mg L-1). From summer to autumn, bottom dissolved oxygen (> 20 m) decreased to reach conditions of hypoxia (1-2 mg L-1) or anoxia; nitrate, nitrite, and phosphates increased (up to 13, 1.7 and 2.2 microM) followed by a last short period of a deep hydrogen sulfide layer (up to 3.1 mg L-1). A back calculation to estimate the input of organic matter in the deep layer showed that 18 g C m-2 y-1 needed to be trapped in the bottom to induce such an oxygen depletion. This period is thought to be unfavorable for bottom infauna in this bay. PMID- 11935904 TI - Abundance and distribution of small infauna in mangroves of Missionary Bay, north Queensland, Australia. AB - To assess the occurrence, spatial distribution and species composition of small infauna on a mangrove shore, core samples were taken along a transect in Missionary Bay at Hinchinbrook Island, north-east Australia. Three sites were arranged within the mangrove forest and one site was located in an adjacent mudflat. The sites were surveyed four times between November 1988 and October 1989. Based on the records from all samples and sites, 39 taxa were identified. Diversity (H') ranged from 1.18 to 2.38. Overall, total abundances of small infauna (retained on a 0.25 mm sieve) reached a mean value of 5,477 ind.m-2, with little variation throughout the transect or over time. Species numbers and diversity were higher in the mudflat than at the mangrove sites. The taxonomic composition changed between the mangrove forest and the mudflat: Oligochaeta were more abundant in mangrove sediments, whereas Polychaeta dominated in the mudflat. Of the polychaetes, Capitellidae were almost restricted to the mangrove sites, Sabellidae were recorded frequently at all sites, and Sigambra parva and Myriochele sp. were confined to the mudflat and the mangrove fringe. These species accounted also for dissimilarities between sites. Multivariate analyses showed a distinct assemblage at the mudflat compared to the mangrove sites. This survey showed that small infauna is an abundant component of mangrove sediments, which has been previously underestimated. PMID- 11935905 TI - Community structure of zooplankton in the main entrance of Bahia Magdalena, Mexico during 1996. AB - The zooplankton community structure, including copepods, euphausiids, chaetognaths, and decapod larvae, was monitored during six circadian cycles using Bongo net (500 microns mesh net) samples from Bahia Magdalena, on the southwest coast of Baja California, Mexico. Samples were obtained during three oceanographic surveys (March, July, and December 1996) to describe the changes in the zooplankton community structure throughout the main mouth of Bahia Magdalena. The zooplankton community structure showed strong changes with a close relation to environmental conditions. During March, a well-mixed water column with low temperature and salinity indicated an influence of the California Current water and local upwelling processes. During July, temperature increased and a wide salinity range was recorded. The stratification of the water column was intense during summer, enhancing the thermocline. The highest temperatures and salinity were recorded in December, related to the presence of the Costa Rica Coastal Current (CRCC). The thermocline deepened as water temperature increased. A typical temperate community structure with low specific richness dominated by Calanus pacificus, Nyctiphanes simplex, and Acartia clausi and high zooplankton biomass (average 9.3 and 5.5 ml 1000 m-3 respectively) during March and July shifted to a more complex tropical community structure with a low zooplankton biomass in December (average 0.37 ml 1000 m-3). The mouth of Bahia Magdalena has a vigorous exchange of water caused by tidal currents. The zooplankton community structure was not significantly different between the central part of Bahia Magdalena and the continental shelf outside the bay for all months. The results suggest a more dynamic inside-outside interaction of zooplankton assemblages than first thought. PMID- 11935906 TI - [Fish larvae association in a Mexican Caribbean bay]. AB - Interannual ichthyoplankton variation, was analyzed in Bahia de la Ascension, Mexico, during December of four consecutive years (1994-1997). A total of 32 families, 35 genera and 21 species of fish larvae were identified. The most abundant fish larvae were the Gobiidae followed by the Callionymidae, Clupeidae and Tetraodontidae. Larval diversity was low when compared with other periods ("dry" and "rainy"). Three spatial associations (internal, medium and external) were found in December 1994 and 1995. In 1996-1997, stations of the inner and outer parts of the bay were mixed. The dominant families characterized most of the faunal associations. Egg density was highest in the external zone of the bay, whereas larvae were most abundant in the inner area. Major factors affecting the fish larval assemblages during December (1994-1997) in Bahia de la Ascension seem to be related to the nursery areas location and to the tropical fishes reproductive period. PMID- 11935907 TI - Impact of logging on a mangrove swamp in south Mexico: cost/benefit analysis. AB - Environmental changes caused by logging in a mangrove swamp were studied in Barra de Tecoanapa, Guerrero, Mexico. Original forest included Rhizophora mangle, Laguncularia racemosa, Avicennia germinans and halophytic vegetation, and produced wood (164.03 m3/ha) and organic matter (3.9 g/m2/day). A total of 3.5 tons of wood per year were harvested from this area. Later, an average of 2,555 kg of maize per planting cycle were obtained (market value of 88 USD). Succession when the area was abandoned included strictly facultative and glycophyte halophytes (16 families, Cyperaceae and Poaceae were the best represented). After logging, temperatures increased 13 degrees C in the soil and 11 degrees C in the air, whereas salinity reached 52 psu in the dry season. These modified soil color and sand content increased from 42.6 to 63.4%. Logging was deleterious to species, habitat, biogeochemical and biological cycles, organic matter production, seeds, young plants, genetic exchange conservation of soil and its fertility, coastal protection, and aesthetic value; 3,000 m2 had eroded as the river advanced towards the deforested area (the cost/benefit analysis showed a ratio of 246:1). There was long-term economic loss for the community and only 30% of the site has recovered after five years. PMID- 11935908 TI - [Stomach content of Spondylus leucacanthus (Bivalvia: Spondylidae) and its relationship with reproduction and phytoplankton in Isla Danzante, Gulf of California]. AB - During 1994 we collected specimens of Spondylus leucacanthus near Isla Danzante, Gulf of California. We measured seawater temperature, and sampled stomach contents and phytoplankton. The reproductive season was assessed through the gonadic index. There is a cool season from December to May and a warm season from June to November. The stomach content was composed of detritus (60%), phytoplankton (30%), and unidentified materials (10%). The highest abundance of phytoplankton cells in the bottom water and the stomach content were in March, April, and May, during the seasonal change of temperature. The gonadic index showed a single peak, confirming that the population has an annual pattern of reproduction. The start of gametogenic phase and maturity in the population coincide with the abundance of phytoplankton. PMID- 11935909 TI - [Mollusc diversity in an Arca zebra (Mollusca: Bivalvia) community, Chacopata, Sucre, Venezuela]. AB - The diversity of a subtidal epifaunal mollusk community was studied from September, 1990 to September, 1991 in Chacopata, Sucre State, Venezuela. There were 40 species (24 bivalves and 16 gastropods). The diversity indexes (H' = 2.087, J' = 0.392, 1/D = 0.528) were low when compared with other tropical zones. Monthly diversity reached its maximum in September, 1990 (1.63 bits/ind.) and July, 1991 (1.60 bits/ind.); minimum diversity occurred in June, 1991 (0.52 bits/ind.). A Log series model applied to species abundance data showed a straight line with a diversity index alpha of 5.56. Of 40 species identified, the turkeywing Arca zebra was dominant (69% in number of individuals and 72% of biomass) followed by Pinctada imbricata, Modiolus squamosus, Chama macerophyla and Anadara notabilis. The predatory snails Phyllonotus pomum, Chicoreus brevifrons and Murex recurvirostris seemed to have trophic relationships with A. zebra. The total mean biomass in wet weight (469.20 +/- 263 g m-2, shell included) was high which indicates that A. zebra, a species with a rapid growth rate, occupies a central role in the assemblage as an efficient filter feeder that converts planktonic food into available biomass, supporting one of the most important fisheries in Venezuela. PMID- 11935910 TI - [Secondary production and condition index in Arca zebra (Mollusca: Vivaldia) in the Gulf of Cariaco, Venezuela]. AB - Production and condition index of a turkeywing (Arca zebra) population were studied from August 1984 to August 1985 in Pariche, Cariaco Gulf, Venezuela. Production was studied through bimonthly collections using a method designed for populations with continuous reproduction and indistinguishable age classes. The population distribution was stable with a mean density of 37 ind.m-2. A production of 131.61 g dry weight m-2 year-1 was calculated from data on density, biomass and weight increase along the year. Highest production was between February 1985 and August 1985 (65%) and concentrated in individuals 60.00-79.95 mm long (45.73%). There was large seasonal variation in the mean monthly condition index (IC = (Ps/Ph) x 100) in the two size classes examined: it reached a maximum in July 1985 (26.84%) and a minimum in January 1985 (16.31%). PMID- 11935912 TI - [Autolytinae, Eusyllinae an Exogoninae (Syllidae: Polychaeta) from Coiba National Park, Panama)]. AB - This is the second paper on the Syllidae (Polychaeta) from hard substrata of the Coiba National Park (Pacific coast of Panama) (the first paper treated Syllinae). On this second paper nineteen species belonging to eleven genera of the subfamilies Autolytinae, Eusyllinae and Exogoninae (Syllidae: Polychaeta) are reported. The samples were collected by SCUBA diving, either by removing 4 kg blocks of dead coral (Pocillopora spp.) or scraping off 25 x 25 cm quadrats of Telesto multiflora or algae (Dyctiota cf. flavellata, Padina cf. durvillaei and another currently unidentified species). They were collected during four expeditions carried out between 1996 and 1998. Four species are newly reported for the Eastern Pacific: Eusyllis lamelligera Marion & Bobretzky, 1875, Exogone exmouthensis Hartmann-Schroder, 1980, Proceraea cf. cornuta (Agassiz, 1863) and Sphaerosyllis (S.) magnidentata Perkins, 1981. Ten species are reported for the first time from the Central America Pacific coast: Amblyosyllis cf. granosa Ehlers, 1897 (sensu Westheide 1974), Amblyosyllis speciosa Izuka, 1912, Autolytus multidenticulatus Westheide, 1974, Exogone naidinoides Westheide, 1974, Grubeosyllis concinna (Westheide, 1974), Odontosyllis fulgurans (Audouin & Milne Edward, 1833), Opisthodonta mitchelli Kudenov & Harris, 1995, Pionosyllis articulata Kudenov & Harris, 1995, Sphaerosyllis (S.) californiensis Hartman, 1966 and Syllides cf. reishi Dorsey, 1978. The family Syllidae is represented in the Coiba National Park by 14% of cosmopolitan species, 14% of circumtropical species, 22% of Atlantic and Pacific species and 25% of Pacific species restricted to warm waters. In samples of dead coral the dominant species are S. armillaris (Muller, 1771) and S. gracilis (Campoy, 1982); in samples of telestaceans Grubeosyllis concinna dominates. PMID- 11935911 TI - [Syllidae (Polychaeta) from Thalassia testudinum beds at Morrocoy National Park (Venezuela)]. AB - A total of 40 species of syllids (Polychaeta: Syllidae) inhabiting Thalassia testudinum beds in Morrocoy National Park, Venezuela, belonging to 13 genera, are reported; 36 species are new for Venezuela: Branchiosyllis exilis, B. oculata, Eusyllis lamelligera, Exogone arenosa, E. breviantennata, E. naidinoides, E. naidina, E. parahomoseta mediterranea, Grubeosyllis clavata, G. heterocirra, Odontosyllis detecta, O. enopla, O. gibba, Opisthosyllis brunnea, Plakosyllis brevipes, Sphaerosyllis bilobata, S. glandulata, S. magnidentata, S. piriferopsis, S. riseri, S. longilamina, S. taylori, Syllis alosae, Syllis riojai, n. comb., S. beneliahui, S. broomensis, S. cornuta, S. corallicola, S. garciai, S. hyalina, S. lutea, S. mexicana, S. prolifera, S. sardai and Xenosyllis scabra. PMID- 11935913 TI - [Growth of Artenia franciscana and A. persimilis (Crustacea, Anostraca) under controlled conditions]. AB - Growth (length) was compared in A. franciscana populations from Utah, United States of America, and Yape, Chile, and A. persimilis (population from Salinas de Hidalgo, Argentina), cultured under controlled conditions (24 degrees C, seawater 35 g/l, fed with Chaetoceros calcitrans). There were significant differences among all stages of A. franciscana (Utah) and A. persimilis, whereas we found significant differences only in two stages of A. persimilis and A. franciscana (Yape); both A. franciscana had similar total lengths in all stages analyzed. Habitat adaptations could explain these results. PMID- 11935914 TI - [Distribution, abundance and population parameters of Diapterus rhombeus (Pisces: Gerreidae) in Laguna de Terminos, Campeche, Mexico]. AB - Diapterus rhombeus dominates the nektonic community of Terminos Lagoon and is associated to zones with submerged vegetation, temperatures between 27 and 33 degrees C and salinity between 25 and 36.5 UPS. A total of 3,024 individuals were collected with a trawl net in 23 sites between September 1997 and September 1998. The length-weight model parameters are a = 0.0098; b = 3.155 with a 96% correlation. The growth parameters are L infinity = 20.4 cm, k = 0.74 and t(o) = 4.207 and the total mortality coefficient is 3.526. Fish length is 2.5-18.0 cm with a mode of 7.5 cm. Recruitment is continuous with a peak from May to August. The species uses several habitats of the lagoon mainly for feeding and growth, reproduction probably occurs in the adjacent continental shelf. Movements between continental shelf and the estuarine system represent a continuous energy flow. PMID- 11935915 TI - Abundance variation of Porichthys margaritatus (Pisces: Batrachoididae) throughout thermic and bathymetric gradients in the central Mexican pacific. AB - Three cruises, one in each of the main stream patterns in the area, were made on the central Mexican Pacific continental slope (May-June 1995, November 1995, and March 1996). Seven trawl sampling sites were defined per cruise, and in each site four bathymetric levels were sampled (20, 40, 60, and 80 m). During the warmest seasons, the highest abundance of Porichthys margaritatus was at 60 m. When the temperature of the bottom water was lower, in March 1996, the highest abundance of P. margaritatus moved towards 40 m. The depths with greatest abundance fluctuated between 18 and 21 degrees C, indicating a narrow temperature preference range. A Gaussian model was used to explain the relation between abundance and temperature. PMID- 11935916 TI - Impacts of Oreochromis mossambicus (Perciformes: Cichlidae) upon habitat segregation among cyprinodontids (Cyprinodontiformes) of a species flock in Mexico. AB - Five species of Cyprinodon in Laguna Chichancanab, Yucatan, Mexico comprise a young species flock whose ecology and evolution has not been thoroughly studied, but whose existence is threatened with extinction. Species flocks evolve in isolated areas where predators and competitors are absent. Since the description of the Chichancanab flock, Oreochromis mossambicus, a species introduced into the lake for which I examined habitat in the 1980's, has become common throughout the basin. I assessed relative abundance of flock species in the lake. examined habitat use and segregation among the three most common flock species and examined the affects of O. mossambicus upon flock species habitat use. Cyprinodon beltrani was the most abundant flock species in 1997, followed by C. maya and C. labiosus; C. verecundus and C. simus were rare. Cyprinodon beltrani was found in shallow water, nearshore, over thick beds of submerged Chara, and little emergent vegetation. Cyprinodon beltrani exhibited diurnal variation in nearshore habitat use. In the field, the habitat use of C. beltrani and O. mossambicus broadly overlapped. In aquarium experiments, three flock species exhibited habitat use segregation and C. beltrani and C. labiosus showed agonistic behaviors that strengthened segregation. Cyprinodon maya differed from C. beltrani and C. labiosus by its greater dispersion of individuals and use of areas higher in the water column. The presence of O. mossambicus caused a shift in habitat use by C. maya and C. labiosus that put these species into habitat occupied by C. beltrani. The presence of introduced species has caused a significant perturbation of the conditions that fomented speciation of the Chichancanab flock 8,000 years ago. PMID- 11935917 TI - [Trophic and reproductive ecology of Trichomycterus calietes and Astrtoblepus cyclopus (pisces: Siluriformes) in Quindio River, Upper Cauca, Colombia]. AB - The trophic and reproductive ecology of catfish (Trichomycterus caliense and Astroblepus cyclopus) was studied in the Quindio River upper Basin, Alto Cauca, Colombia. The pH was neutral, water oxygen content high (8.4 ppm) and temperature in the habitats was 18.63 degrees C; both species are nonmigratory and sympatric with four other fish species. The ovaries mature primarily between May and September in T. caliense; between December and May in A. cyclopus. The mean size at maturity is 8.3 cm (standard length) in T. caliense and 6.0 cm (standard length) in A. cyclopus; the sex ratio is 1:1 in T. caliense (X2 = 3.4, P > or = 0.05) and in A. cyclopus (X2 = 1.44, P > or = 0.1); the fecundity is low (191 and 113 oocytes respectively) and the eggs are small (1.5 and 2.39 mm respectively). The fishes are insectivorous and specialize in Coleoptera, Diptera and Trichoptera; Spearman Rank Correlation Coefficients (rs = 0.464) indicated that there are differences (T = 2.5148, P < 0.01) between their diets; both taxa did not agree with the expected trophic habits for sympatric species that are morphologically similar and related in the same trophic level. PMID- 11935918 TI - Cetacean strandings in Costa Rica (1966-1999). AB - Cetacean strandings in Costa Rica are reported for a period of 33 years, with a total of 35 strandings, 13 species and 247 individuals involved. The vast majority of documented strandings occurred on the Pacific coast and correspond to single individuals (32 and 28 strandings respectively). The highest stranding number was in the period from 1990 to 1999 (n = 24). Physeter catodon (cachalot or sperm whale) is the species with the highest frequency of strandings (n = 8) and the family Delphinidae has the majority of species (n = 8) and strandings (n = 22). No other general tendencies were determined with the existing data. PMID- 11935919 TI - The attraction of Zonocerus variegatus (Orthoptera: Pyrgomorphidae) to different types of lure. AB - Zonocerus variegatus is a common grasshopper in parts of west and equtorial Africa. The distribution in Nigeria extends from the lowland rainforest zone to the savannah in the north. The influence of lure on the behaviour of grasshopper inside cages (120 insect per cage) was investigated. Nymphs and adults of Zonocerus variegatus responded positively to intact leaves, crushed leaves and inflorescence of the common compositae weed Chromolaena odorata inside muslin bags, and intact plants. There were significant differences in the attraction recorded for starved nymphs, fed nymphs and starved adults. Attraction was more to intact leaves and is by olfaction. The increase in the attraction of starved nymphs is time dependent. Attraction to plant parts ceased after the plants were dried for 24 and 48 hours at room temperature and when plants were placed in transparent polythene bags. Gut motility and gut activity were higher during the day than at night. Nymphs, adults and egg pods placed separately inside muslin bags were not attractive to adults or nymphs. PMID- 11935920 TI - Effect of food plants on the volume of repellent secretion obtained in adult Zonocerus variegatus (Orthoptera: Pyrgomorphidae). AB - The volume of secretion obtained from adult Zonocerus variegatus (Orthoptera: Pyrgomorphidae) was influenced by the type of food plants. Insects fed on leaves of cassava Manihot esculenta, bitter leaves Vernonia amygdalina, and a mixture of M. esculenta and Acalypha wilkesiana gave a good volume of secretion while Chromolaena odorata, Elaeis guinensis, Aspilia africana and Citrus sinensis did not favour secretion production. No significant difference was recorded in the volume of secretion obtained from Z. variegatus from the two seasons irrespective of the food plant. Similarly, food plants gave no significant difference on the volume of secretion between the two seasons. PMID- 11935922 TI - [Effect of human blood feeding on the fecundity, fertility and biological cycle of Rhodnius prolixus (Hemiptera: Reduviidae)]. AB - The effect of several human blood fractions artificially fed to Rhodnius prolixus Stal 1859 on oviposture (fecundity), egg-hatching (fertility) and life cycle was observed. Specimens fed on man's blood were more fecund than those fed with woman's blood. There were no significant differences in fertility related to host sex. The nymphal development time and number of feedings to molt to the following instar were estimated. Animals fed only on blood plasm did not finish nymphal development, while those fed only blood red cells ended their life cycle in the third nymphal instar. Total life cycle lasts 129 days in individuals fed with whole blood. PMID- 11935921 TI - Sylvatic focus of American Trypanosomiasis in the State of Morelos, Mexico. AB - Wild vectors and reservoir hosts of Trypanosoma cruzi were surveyed from February 1993 to June 1994 in Ticuman (18 degrees 46'N, 99 degrees 07'W), Mexico (Deciduous Tropical Forest). Direct faeces examination showed that 87% of Triatoma pallidipennis hosted the parasite; T. cruzi forms were present in cultures inoculated with faeces of fifty 67% triatomine bugs and thirty CD-1 strain mice (10 d old) inoculated (peritoneum) with faeces of positive insects T. cruzi amastigotes were found in heart 67%, kidneys 47%, liver 80%, lungs 50%, oesophagus 60%, skin 23%, spleen 73% and stomach 60%. T. cruzi was isolated by direct blood examination from seven 21% chiropterans and five 38% rodents and T. cruzi forms were present in cultures inoculated with blood of twenty-three 68% chiropterans and seven 54% rodents and T. cruzi amastigotes were seen in the kidneys of one 3% chiropterans and four 31% rodents and only in one Pteronotus parnellii mexicanus, organisms were seen in skin 2%. There was no association between organs and T. cruzi infection (p > 0.05). PMID- 11935923 TI - [Feeding and defecation of Rhodnius (hemiptera: Reduviidae) fed human blood]. AB - Feeding and defecation behavior of Rhodnius prolixus Stal, 1859, R. robustus Larrousse, 1927, R. neivai Lent, 1953 and R. pictipes Stal, 1872, artificially fed on human blood, were studied under laboratory conditions. In all species, first instar nymphs did not defecate in the first 30 minutes after feeding. R. pictipes did not accept artificial feeding but fed directly on humans. Nymph and adult R. prolixus had a higher defecation index (DI) than other species; third instar nymphs had the highest DI = 1.62. In all instars, most individuals accepted the food in 3 Pounds minutes and finished feeding in less than 15 minutes. PMID- 11935924 TI - Within-tree distribution of Ecdytolopha torticornis (Lepidoptera: Tortricidae) oviposition on macadamia nuts. AB - Vertical distribution of eggs of the macadamia nutborer Ecdytolopha torticornis Meyrick (Lepidoptera: Tortricidae) and its preference of oviposition sites within and between macadamia cultivars were studied in Turrialba, Cartago, Costa Rica, in 1992 (N = 6,939). E. torticornis eggs were found throughout the foliar parts of the tree, but fewer eggs were laid in the crown top than in the mid or lower crown. Differences in the horizontal distribution of the eggs were not significant, albeit more eggs were found in the outer positions. The numbers of eggs found within the crowns of different clones were similar, implying that the nutborer has no preference for a particular cultivar. PMID- 11935926 TI - Morphological variation in the tropical anole, Anolis casildae (Squamata: Polychrotidae). AB - We describe morphological variation (scalation and coloration) observed among eight individuals of the Panamanian lizard species Anolis casildae. This variation was not observed in the holotype and aids in identification of this recently described species (originally described on the basis of a single, male specimen). This species occurs only in the Reserva Forestal Fortuna (Chiriqui Province) and the adjacent Bosque Protector Palo Seco (Bocas del Toro Province) in western Panama. Anolis casildae can be distinguished from all other Panamanian anole species via six features: (1) two enlarged superciliary scales (the first larger than the second); (2) an anterior nasal scale in contact with the rostral scale or separated from the rostral by one scale; (3) 6-8 sublabial scales to the center of the eye; (4) 3-4 scales between the supraobital semicircles; (5) unique coloration (4-6 oblique brown bands interspersed by blue-outlined yellow patches; dewlap is a dirty cream color with broad yellow scale rows irregularly interspersed with smaller emerald green scales) and (6) A. casildae occurs from 1,050 to 1,400 m in the Cordillera Central. We also compare our natural history observations of A. casildae to a similar large anole, A. frenatus, a species which we believe A. casildae to be closely related. PMID- 11935925 TI - Distribution, habitat and behavior of grasshopper sparrows, Ammodramus savannarum (Passeriformes: Emberizidae) in northeastern Nicaragua. AB - During March and April of 1996, I made field observations of the sedentary subspecies of grasshopper sparrow (Ammodramus savannarum cracens), in 600 points of the pine savannas of northeastern Nicaragua. Isolated individuals were found in the humid depressions, but breeding populations were located exclusively in areas that had suffered a recent fire. Territorial behavior varied in intensity apparently as a function of territory size: the most aggressive males were those trying to defend smaller territories in populations close to Miskito villages, where most of the fires occur. In contrast to what is happening in other parts of Central America, the Nicaraguan grasshopper sparrow may be indirectly protected from extinction by the Miskito's traditional fire practices. PMID- 11935927 TI - A taxonomic reevaluation of Phrynops (Testudines: Chelidae) with the description of two new genera and a new species of Batrachemys. AB - Relationships among turtle species loosely categorized within the South American genus Phrynops are explored. Three once recognized genera (Batrachemys, Mesoclemmys and Phrynops) that were demoted to subgenera, and then synonymized with Phrynops, are demonstrated to warrant full recognition based on morphometric analysis, skull osteology, and mitochondrial and nuclear gene sequencing. Mesoclemmys is resurrected from the synonymy of Phrynops as a monotypic genus including M. gibba. The genus Rhinemys, previously a synonym of Phrynops, is resurrected for the species R. rufipes. Ranacephala gen. nov. is described to include the species R. hogei. The genus Batrachemys is resurrected from the synonymy of Phrynops and includes B. dahli, B. nasuta, B. raniceps, B. tuberculata, and B. zuliae. The taxon vanderhaegei is placed in Bufocephala gen. nov. The genus Phrynops is redefined to include the taxa P. geoffroanus, P. hilarii, P. tuberosus, and P. williamsi. Cladistic analysis of morphological data supports this taxonomy. A new species of Batrachemys is described from the western Amazon region, and is distinguished by having facial markings in juveniles, a relatively wide head, and a flattened shell. The new species, B. heliostemma sp. nov., is sympatric with and most similar to the recently resurrected form Batrachemys raniceps in the upper Amazonian region of Peru and adjacent Brazil, Ecuador, Venezuela, and Colombia. Lastly, morphometric data from living and museum specimens of all species of Batrachemys are presented. PMID- 11935928 TI - Pre-clinical performance comparing intraoral film and CCD-based systems. AB - PURPOSE: The purpose of this study was to compare dental hygiene student radiographic performance using film and charge-coupled device (CCD) systems. Specific objectives were to compare the 1) number of technique errors, 2) error type, 3) retake frequency, and 4) learning experience using both systems. METHODS: Twenty-eight subjects exposed two 18-projection full series, one film based, and one CCD-based system, on a manikin. One examiner, using standardized image quality criteria, evaluated all images. Each subject was assigned to one of four focus groups. Quantitative data were analyzed using ANOVA and Wilcoxon tests. Emic expressions and percentages were used to report qualitative data with themes supported by performance data. RESULTS: A greater number of technique errors occurred using the CCD system than the film system (p < 0.0001), with vertical angulation being the most common error (53%). Using the film-based system, 66% of the errors were due to incorrect horizontal angulation. Subjects exposed an average of 10 retakes per series using the CCD versus 3 retakes using film. Forty-four percent of the subjects felt their most common CCD errors were film placement and vertical angulation compared to 59% who believed horizontal angulation was the main technique error when using film. Difficulty placing the CCD sensor intraorally was expressed by 74%. The CCD system was reported to be beneficial for teaching theory (67%), with 59% preferring film for learning the psychomotor skill. CONCLUSION: In pre-clinical situations, the CCD system may be an effective tool for teaching technique. Radiographic errors are dependent on the type of image receptor used. PMID- 11935930 TI - Happy 75th anniversary to the journal of dental hygiene. PMID- 11935929 TI - Use of local anesthesia by dental hygienists who completed a Minnesota CE course. AB - PURPOSE: This study assessed the use of local anesthesia administered by dental hygienists who completed continuing education (CE) courses in Minnesota in 1996. The aims of the study were to: evaluate the use of local anesthesia by these participants; the level of effective injections; the value the dental hygienists placed on this skill; and to provide data in areas less studied, such as injection techniques utilized, and the incidence of complications. METHODS: A pretested, 34-item questionnaire; cover letter; and return self-addressed, stamped envelope were sent in 1997 to dental hygienists (N = 355) who completed a CE course in local anesthesia offered by Minnesota dental hygiene programs between January and September 1996. A reminder postcard was sent to nonrespondents two weeks after the initial return date. The survey assessed the administration experience of dental hygienists including implementation of this skill, frequency of administration, types of clients receiving local anesthesia, treatments requiring local anesthesia, choice of injection techniques, level of successful anesthesia, complications observed, and value to practice. Data were analyzed using JMP version 3.1, Pearson's chi-square and one-way ANOVA. RESULTS: The response rate was 77.7% (N = 273). Ninety-five percent of the dental hygienists who responded reported delegation of local anesthesia to them by employers. A majority of respondents (64%) implemented this new skill within one week following completion of course work. Dental hygienists in periodontal specialty practice provided local anesthesia to adult clients more frequently than those in general practice. Respondents used nerve block techniques more frequently than supraperiosteal or infiltration techniques. In general, respondents reported high success rates in achieving adequate anesthesia with a very low incidence of complications. Dental hygienists most frequently administered local anesthesia for root planing or debridement (92%). A majority of respondents also administered local anesthesia to adults and children who were treated by the dentist. The frequency of administrations was strongly related (P = 0.00) to value placed on this skill and positive outcomes to their practice. CONCLUSIONS: Dental hygienists who completed required CE course work in local anesthesia, provided safe, effective injections and reported this service to be of value to their practice. These data support the incorporation of local anesthesia into dental hygiene curricula to prepare future practitioners to provide this service to clients. Also, dental hygienists working in states not currently allowing this service, should consider these results when proposing change to their state practice acts. PMID- 11935931 TI - Health, mental well-being, and musculoskeletal disorders: a comparison between Swedish and Australian dental hygienist. AB - PURPOSE: This study was conducted to compare Swedish and Australian dental hygienist in regards to working conditions, health, and mental well being and to further develop a health model for this profession. METHODS: Questionnaires (a 275-item instrument) were sent to all 376 Australian dental hygienist and a random sample of 26% (n=575) of Swedish dental hygienist. Country-specific factors--such as work characteristics and life style--and their relationships to self-related health and mental well being were analyzed. Interaction effects of age times country were studied by two-factor ANOVA. Discriminant functions were used to predict country and age group memberships, and multiple logistic regression modeling for a health model was tested. RESULTS: The response rate was 86% (N=495) in Sweden and 71% (N=254) in Australia. Work-related musculoskeletal disorders, professional ambition, and demands from difficult patients were the factors most different for the two countries--generally the values were higher for Australia than Sweden. Arthritic hands/loss of sensitivity in the arm-hand and intellectual job satisfaction were found more often in dental hygienist more than 42 years old. than in dental hygienist ages under 42 years old. Professional ambition was higher for those under 42. Working 25 hours or more per week with clinical work, engaging in active leisure, being under age 42, and high management support at work, relate to good general health. Work-family overload was associated with lower general health, while higher mental well-being was related to high perception of personal mastery and high management support. There also was a correlation between lower mental well-being was and high work-family overload, work efficiency, and practicing in Australia. Work-family overload, scaling procedures, work relations, practicing in Australia, and being under 42 years of age were associated with more musculoskeletal disorders. CONCLUSION: An earlier hypothetical work and health model, on which this study was based, was expanded with these factors: country-specific work setting, age, and work relations. Australian dental hygienist and more musculoskeletal disorders and experienced lower mental well being related to organizational work factors than did Swedish dental hygienist. The results suggest a need to consider sociodemographic, occupational, and cultural aspects in future studies of work, health, and well being. PMID- 11935932 TI - Short report on antimicrobials in periodontal maintenance. PMID- 11935935 TI - Resorbable polymeric sponges as drug carriers. Part 2. Gelatin sponge. AB - The aim of the research is to obtain a porous and bioresorbable gelatin matrix, which could be used as a drug carrier. The research presented in the paper is a continuation of the work carried out earlier. PMID- 11935934 TI - Diet and nutrition implications for oral health. AB - Dental hygienists are in a unique position to provide both oral hygiene instruction and basic nutrition information related to oral health. Many clients may not be aware of the effects of diet and nutritional status on the development and maintenance of a healthy mouth and caries-free teeth. It is vitally important for dental hygienists to perform basic nutrition screening to assess clients' dietary habits for potential risk factors for dental caries and periodontal disease, and to look for potential signs of nutritional deficiencies or nutrition related problems in clients with chronic diseases or conditions. Collaboration between dietetics professionals and oral health care professionals is essential in identifying, educating, and treating oral health problems related to nutrition. Such partnerships will encourage improved levels of oral health care. The purposes of this course are to review the links between nutrition and oral health and to discuss how oral health care professionals can integrate nutrition counseling aimed at improving the oral health of their clients into their practices. PMID- 11935933 TI - Incorporation of evidence-based principles in baccalaureate and nonbaccalaureate degree dental hygiene programs. AB - PURPOSE: The purpose of this descriptive study was to determine the curricular utilization of Evidence-Based (EB) philosophies in dental hygiene education in both baccalaureate and nonbaccalaureate U.S. dental hygiene programs. METHODS: Data were gathered via a 1999 survey of all 235 U.S. dental hygiene program directors. The 21-question survey included one open-ended and 20 closed-ended items. Initially, the survey was pilot tested using a convenience sample of seven U.S. dental hygiene program directors. A final, revised survey was mailed to the cohort population. A response rate of 71% (N = 167) was achieved after two mailings. RESULTS: The demographic results of this study revealed the majority of respondents (77%) were from nonbaccalaureate dental hygiene programs, while the remainder (23%) were from baccalaureate degree programs. Respondents were asked if and to what degree the fundamentals of research were taught in their curriculum. Baccalaureate programs' responses indicated a much greater degree of utilization. Sixty-two percent of baccalaureate respondents provide a separate course on research, and 32% teach research as a portion of another dental hygiene course. However, 3% of the responding baccalaureate programs reported that no research was included in the curriculum. These results demonstrate an overall high utilization of research by baccalaureate curricula. In comparison, nonbaccalaureate programs' responses show a lesser degree of research utilization when compared with the baccalaureate programs. Only 8% of nonbaccalaureate programs had a separate research course. The majority, 80%, of nonbaccalaureate respondents reported that they teach research as a portion of another dental hygiene course. Finally, 10% of the nonbaccalaureate degree respondents reported teaching no research. Overwhelmingly, both baccalaureate and nonbaccalaureate students received formal orientation in the use of library--100% and 84%, respectively. Ninety-seven percent of baccalaureate and 82% of nonbaccalaureate programs provided students with formal instruction in using both literature indices and databases. To a lesser extent, both baccalaureate and nonbaccalaureate programs introduced students to the Internet and encouraged them to use it for conducting literature searches--83% and 78%, respectively. In fact, seventy-four percent of baccalaureate programs and 68% of nonbaccalaureate programs reported formally teaching the evaluation of information retrieved from the Internet. Both baccalaureate and nonbaccalaureate programs similarly indicated teaching the evaluation of research findings for validity, reliability, and clinical importance. To a lesser extent, 86% of baccalaureate and 61% of nonbaccalaureate programs reported that they encourage students to make EB recommendations to patients and teach students how to apply EB findings to clinical situations. CONCLUSION: The findings of this descriptive study indicate both baccalaureate and nonbaccalaureate degree programs incorporate some aspects of an EB philosophy into the curriculum. Survey results reveal baccalaureate degree programs incorporated research and taught the use of library facilities, journal indices, and electronic databases. In addition, baccalaureate degree programs also emphasized and encouraged the application of critically appraised evidence into practice. The nonbaccalaureate respondents utilized the library facilities, journal indices, and electronic databases to a slightly lesser extent than their baccalaureate counterparts. The nonbaccalaureate respondents also demonstrated less application of EB findings to clinical situations, including actual patient treatment. PMID- 11935937 TI - Preliminary assessment of selected methacrylic acid--acrylic acid copolymers as factors buffering triethanolamine interacting with artificial skin sebum. AB - To allow cleaning of sebum ducts for acne prophylaxis the authors suggest use of triethanolamine buffered with anionic polymers of acrylic and methacrylic acid. In these preliminary researches addition of Eudragit L-100, Eudragit L-100-55 or Eudragit S-100 implicates decrease of pH of 1.49% triethahnolamine solution from 10.06 even to 7.20. Triethanolamine penetrates into the artificial skin sebum, gains hydrophylic properties, and reacted sebum increase in volume allowing mechanical remove of lipid bed. The highest triethanolamine penetration and the highest volume increase were observed after application of Eudragit S-100 complexes. Eudragit L-100 and L-100-55 proved milder action. These complexes may be applied for further examinations on acne prevention. PMID- 11935936 TI - [Gravitational osmotic pressure effect for a series of flat polymer membranes positioned horizontally]. AB - In this paper the results of study pressure graviosmotic effect for a double membrane osmotic-diffusive cell, in which series of two (Ml and M(r)), microporous and symmetrical flat polymeric membranes (Nephrophane and Cellulose IMP-1) separate three compartments (l, m, r) containing the heterogeneous and binary (aqueous glucose or ethanol solutions) or ternary (glucose solutions in 0.75 mole.l-1 aqueous ethanol solution or ethanol solutions in 0.1 mole.l-1 aqueous glucose solution) non-ionic solutions. In this system the solution concentrations fulfill the condition Ckl > Ckm > Ckr. The inter-membrane compartment (m) consists of the infinitesimal layer of solution. The volume of compartment m and external compartment (l and r) fulfill the conditions Vm-->0 and Vl = Vr-->infinity respectively. The calculations of pressure graviosmotic effect for configurations A and B of the double-membrane osmotic-diffusive cell were elaborated. In configuration A solution was placed in compartment below membrane M(r) and water above membrane Ml. In configuration B solution was placed in compartment above membrane Ml and water below membrane Ml. These calculated results are interpreted in terms of the convective instability that increases the diffusive permeability coefficients of complexes: concentration boundary layers (membrane Ml or M(r)) concentration boundary layer. PMID- 11935938 TI - [Streaming gravity-diffusive effect for a series of two flat polymeric membranes oriented horizontally]. AB - In this paper the results of study flux gravidiffusive effect for a double membrane osmotic-diffusive cell, in which series of two (Ml and M(r)), microporous and symmetrical flat polymeric membranes (Nephrophane and Cellulose IMP-1). These membranes separate three compartments (l, m, r) containing the heterogeneous and binary (aqueous glucose or ethanol solutions) or ternary (glucose solutions in 0.75 mole.l-1 aqueous ethanol solution or ethanol solutions in 0.1 mole.l-1 aqueous glucose solution) non-ionic solutions. The solution concentrations fulfil the condition Ckl > Ckm > Ckr. The inter-membrane compartment (m) consists of the infinitesimal layer of solution. The volume of compartment m and external compartment (l and r) fulfill the conditions Vm-->0 and Vl = Vr-->infinity respectively. The study of flux gravidiffusive effect for configurations A and B of the double-membrane osmotic-diffusive cell were elaborated. In configuration A solution was placed in compartment below membrane M(r) and water above membrane Ml. In configuration B solution was placed in compartment above membrane Ml and water below membrane Ml. These results are interpreted in terms of the convective instability that increases the diffusive permeability coefficients of complexes: concentration boundary layers/membrane Ml or M(r)/concentration boundary layer. PMID- 11935939 TI - [Streaming gravity-osmotic effect for a series of two flat polymeric membranes oriented horizontally and ternary non-ionic solutions]. AB - In this paper the results of study flux graviosmotic effect for a double-membrane system, in which two (Ml and M(r)), microporous and symmetrical flat polymeric membranes (Nephrophane and Cellulose IMP-1) separate three compartments (l, m, r) containing the heterogeneous and binary (aqueous glucose or ethanol solutions) or ternary (glucose solutions in 0.75 mole.l-1 aqueous ethanol solution or ethanol solutions in 0.1 mol.l-1 aqueous glucose solution) non-ionic solutions. In this system the solution concentrations fulfill the condition Ckl > Ckm > Ckr. The inter-membrane compartment (m) consists of the infinitesimal layer of solution. The volume of compartment m and external compartment (l and r) fulfill the conditions Vm-->0 and Vl = Vr-->infinity respectively. The calculations of flux graviosmotic effect for configurations A and B of the double-membrane osmotic diffusive cell were elaborated. In configuration A solution was placed in compartment below membrane M(r) and water above membrane Ml. In configuration B solution was placed in compartment above membrane Ml and water below membrane Ml. These calculated results are interpreted in terms of the convective instability that increases the diffusive permeability coefficients of complexes: concentration boundary layers/membrane Ml or M(r)/concentration boundary layer. PMID- 11935940 TI - The preliminary evaluation of HAP + TCP composite material biodegradation after implantation in muscular tissue of rats. AB - Ceramic biomaterials based on calcium phosphates have a special position among modern implantation material in osteosurgery. Non-reabsorbable hydroxyapatite (HAP) and reabsorbable tricalcium phosphate are the most popular calcium phosphate ceramics. The appropriate ratio of these two compounds should result in forming the gradually reabsorbable implants, overgrowing with the bone tissue which mechanical strength should not be negatively affected. The aim of this work was to evaluate a local tissue reaction and the HAP + TCP composite resorption rate as compared with HAP, after implantation in a muscle tissue of rats. On the basis of carried macroscopic and microscopic evaluations, it can be stated that the new HAP + TCP composite had high biocompatibility and were gradually reabsorbed. This enables faster overgrowing the implant with tissue. PMID- 11935941 TI - Impact of daily food choices on health promotion and disease prevention. PMID- 11935942 TI - Beneficial effects of eicosapentaenoic acid on endothelial vasodilator functions in animals and humans. PMID- 11935943 TI - omega 3 polyunsaturated fatty acids, inflammation and immunity. PMID- 11935944 TI - Effect of eicosapentaenoic acid on the suppression of growth and metastasis of human breast cancer cells in vivo and in vitro. PMID- 11935946 TI - Effect of omega 3 polyunsaturated fatty acid ingestion on bone metabolism and osteoporosis. PMID- 11935945 TI - Biochemical and molecular actions of fatty acids in bone modeling. PMID- 11935947 TI - Dietary fat subtypes and obesity. PMID- 11935948 TI - Studies on the role of dietary diacylglycerols in human nutrition. PMID- 11935949 TI - omega 3 fatty acids and human chylomicron metabolism. PMID- 11935950 TI - Production and release of docosahexaenoic acid by differentiated rat brain astrocytes. PMID- 11935951 TI - Lysophosphatidylcholine as a carrier of docosahexaenoic acid to target tissues. PMID- 11935952 TI - New developments in alpha-linolenate metabolism with emphasis on the importance of beta-oxidation and carbon recycling. PMID- 11935953 TI - Evolutionary aspects of diet and essential fatty acids. PMID- 11935954 TI - Polyunsaturated fatty acid metabolism during lactation. PMID- 11935955 TI - Differences in the regulation of the biosynthesis and esterification of 20- versus 22-carbon polyunsaturated fatty acids. PMID- 11935956 TI - Cloning of rat delta 6-desaturase and its regulation by dietary eicosapentaenoic or docosahexaenoic acid. PMID- 11935957 TI - 2-arachidonoylglycerol: an endogenous cannabinoid receptor agonist. PMID- 11935958 TI - Role of N-acylethanolamines in cell signaling. PMID- 11935959 TI - Alkaline and acid amidases hydrolyzing anandamide and other N-acylethanolamines. PMID- 11935960 TI - When and where are N-acylethanolamine phospholipids and anandamide formed? PMID- 11935961 TI - Bioactions and synthesis of a novel lipid mediator, cyclic phosphatidic acid. PMID- 11935962 TI - Phenolics from olive oil and its waste products. Biological activities in in vitro and in vivo studies. PMID- 11935963 TI - Isomeric fatty acids: their physiological significance and health implications. PMID- 11935964 TI - Transgenic production of long-chain polyunsaturated fatty acids. PMID- 11935965 TI - Recent development of health foods enriched with DHA, EPA and DPA in Japan. PMID- 11935966 TI - Effect of omega 6 and omega 3 fatty acid supplementation on rat tissue lipid peroxidation. PMID- 11935967 TI - alpha-linolenate-deficiency-induced alterations in brightness discrimination learning behavior and retinal function in rats. PMID- 11935968 TI - Seafood consumption and homicide mortality. A cross-national ecological analysis. PMID- 11935969 TI - Effect of docosahexaenoic acid on hostility. PMID- 11935970 TI - Synaptic functions and synaptic membrane lipids in the aging brain. PMID- 11935971 TI - Methodological considerations in clinical studies of omega 3 fatty acids in major depression and bipolar disorder. PMID- 11935972 TI - Docosahexaenoic acid and cerebral evolution. PMID- 11935973 TI - Effect of DHA oil supplementation on intelligence and visual acuity in the elderly. PMID- 11935974 TI - Electrophysiologic basis for the antiarrhythmic and anticonvulsant effects of omega 3 polyunsaturated fatty acids. PMID- 11935975 TI - alpha-linolenic acid in the prevention of cardiovascular diseases. PMID- 11935976 TI - Fish oil fatty acids beneficially modulate vascular function. PMID- 11935977 TI - omega 3 fatty acids. From eskimos to clinical cardiology--what took us so long? PMID- 11935978 TI - Legal reform: the gathering storm. PMID- 11935979 TI - Malpractice crises. PMID- 11935980 TI - Compliance issues for physicians and physician organizations. PMID- 11935981 TI - Keeping our helmet laws--who is the best teacher? PMID- 11935982 TI - Flatback deformity--diagnosis and treatment. PMID- 11935983 TI - Health economics and technology reimbursement. Payer impact. PMID- 11935984 TI - Discriminating between supraventricular and ventricular tachycardias from EGM onset analysis. PMID- 11935985 TI - Support vector black-box interpretation in ventricular arrhythmia discrimination. PMID- 11935986 TI - Interpreting ECG data by integrating statistical and artificial intelligence tools. PMID- 11935987 TI - The principles of software QRS detection. PMID- 11935988 TI - Finding coordinated atrial activity during ventricular fibrillation using wavelet decomposition. PMID- 11935989 TI - Poincare mapping for detecting abnormal dynamics of cardiac repolarization. PMID- 11935990 TI - Improved estimation of left ventricular hypertrophy. PMID- 11935991 TI - Effects of radiofrequency ablation on monophasic action potentials. PMID- 11935992 TI - Studying cardiac contractility change trend to evaluate cardiac reserve. PMID- 11935993 TI - Electric field interactions between closely abutting excitable cells. . PMID- 11935994 TI - EMF cancer scares: epidemiology versus body power. PMID- 11935996 TI - Science in medicine--an elusive notion. PMID- 11935995 TI - Patterning the cellular microenvironment. PMID- 11935997 TI - Effects of an oral multivitamin-mineral combination on stress. PMID- 11935998 TI - Drug interactions--'natural' not necessarily synonymous with 'safe'. PMID- 11935999 TI - HIV, morality and human rights. PMID- 11936000 TI - HIV infection in northern Mozambique. PMID- 11936002 TI - Revolutionary Lodox's international debut. PMID- 11936001 TI - Another mother's milk saves AIDS babies. PMID- 11936003 TI - A tragic and lucrative growth industry. PMID- 11936004 TI - SAMA board tackles point of care services. PMID- 11936006 TI - Rising cost of medical AIDS. PMID- 11936005 TI - Profession to examine prison health. PMID- 11936007 TI - Medscheme's new life assurance. PMID- 11936008 TI - Human rights and ethical guidelines on HIV. PMID- 11936009 TI - Practice marketing techniques. II: Public relations. PMID- 11936010 TI - Psychoimmunology and HIV/AIDS. PMID- 11936011 TI - Removing lumps and bumps in general practice. PMID- 11936012 TI - CT scanning--essential for conservative management of paediatric blunt abdominal trauma. PMID- 11936013 TI - Intradermal BCG vaccination--adverse reactions and their management. PMID- 11936014 TI - The days of miracle and wonder. PMID- 11936015 TI - The ethics of immunisation. PMID- 11936016 TI - Use of goat's milk in cow's milk-allergic infants. PMID- 11936017 TI - Management of acute respiratory infection in an ambulatory setting. PMID- 11936018 TI - Endoscopic aqueductoplasty and stenting--report of initial experience. PMID- 11936019 TI - Safety and immunogenicity of TETRActHIB (a vaccine combining DTP vaccine and Haemophilus influenzae type B conjugate vaccine) administered to infants at 6, 10 and 14 weeks of age. AB - The safety and immunogenicity of TETRActHIB (a vaccine combining diphtheria and tetanus toxoids-pertussis vaccine (DTP) with Haemophilus influenzae type b (Hib) conjugate vaccine (polyribosyl ribitol phosphate conjugated to tetanus protein) (PRP-T)) was assessed in 131 Cape Town infants immunized at 6, 10 and 14 weeks of age. Serological responses to all component antigens were measured before the first dose and at 18 weeks of age. In addition, anti-PRP antibodies were measured at 9 and 18 months of age to determine long-term immunogenicity. The vaccine was well tolerated by infants and no significant side-effects were reported. Responses to Hib at 18 weeks of age were good in that most infants achieved a level of anti-PRP antibodies > or = 0.15 microgram/ml, indicative of short-term protection, and 70% achieved a level > or = 1 microgram/ml, indicative of long term protection. The proportions of children with protective levels > or = 0.15 microgram/ml and > or = 1 microgram/ml were similar at 9 and 18 months of age, i.e. approximately 75% and 45%, respectively. Responses to tetanus and diphtheria toxoids were excellent and all infants achieved protective serological levels. Responses to pertussis were moderate in that approximately 65% achieved 'protective' serum levels of pertussis agglutinins, i.e. titres > or = 320. In conclusion, this study has shown that the DTP/PRP-T vaccine is safe, immunogenic and well tolerated in infants immunised at 6, 10 and 14 weeks of age. TETRActHIB is therefore suitable for inclusion in the World Health Organisation Expanded Programme on Immunisation (WHO EPI) schedule. PMID- 11936020 TI - Cigarette smoking in an adolescent psychiatric population. AB - OBJECTIVE: To examine the relationship between cigarette smoking and psychiatric symptomatology in an outpatient psychiatric population of adolescents. METHOD: A retrospective analysis was done of 934 patient charts at an outpatient psychiatric centre. RESULTS: 48.4% of the psychiatric sample reported regular smoking behaviour, which is substantially more than the 18.1% prevalence found in a local epidemiological study. In comparing smokers and non-smokers within the psychiatric sample, it was noted that smokers were significantly younger and scored somewhat higher on depression rating scales than non-smokers. A logistical regression, using quasi-Newton estimation, was chosen as the most suitable statistical method for building a classificatory model of smoking. Two continuous variables, age and the Hamilton depression score, along with 39 discrete variables, were chosen for modelling purposes. Model building was conducted in a hierarchical fashion, starting with demographic variables, the variable selection being controlled by using chi-square tests of model differences. A predictive model of smoking with nine variables was finally selected. CONCLUSIONS: As a whole the results support the strong association between smoking and psychiatric problems, but in this adolescent sample smoking is more likely to be part of a general risk-taking behaviour pattern than an attempt to medicate depression. Anti-tobacco campaigns that highlight the risks of smoking are therefore open invitations for adolescents to take up the habit. PMID- 11936021 TI - Trabecular bone density in premenopausal rheumatoid arthritis patients. AB - OBJECTIVE: This study was undertaken to compare trabecular bone mineral density (BMD) in premenopausal rheumatoid arthritis (RA) patients and normal age-matched controls. METHOD: A protocol was designed to record age, duration of disease, use of corticosteroids (CS) and/or slow-acting antirheumatic drug (SAARD) therapy together with duration of such therapy. BMD was measured using the Hologic QDR 1,000 dual energy X-ray absorptiometer. The first four lumbar vertebrae and the left femur were measured in 56 RA patients and 165 controls. Height and weight were measured. Comparisons were made between RA patients and controls, as well as between subgroups of RA patients based on CS therapy. RESULTS: Patients with RA had significantly lower BMD (P < 0.05) at all the sites than the normal controls. The mean duration of RA at the time of study was 60 months (standard deviation 58 months). Thirteen RA patients had used CS in doses less than 10 mg daily for 6 months or longer (mean 19 months), while 25 patients had been on SAARD for an excess of 6 months (mean 23 months). The CS-treated patients had significantly lower BMD than untreated subjects at the femoral neck and inter-trochanteric region (P < 0.05), but not at the lumbar spine. However, when compared with normal controls, the CS-treated subgroups had significantly lower BMD at the lumbar spine and all femoral areas. Trochanteric BMD was the best determinant of the RA group, with a sensitivity of 65% and specificity of 77%. The positive predictive value was 16%, while the negative predictive value was 10%. Using Bayes' theorem, the prevalence of osteopenia in RA was found to be 6%. CONCLUSION: We conclude that generalised bone loss is a systemic feature of RA and that loss at the spine and femur may be aggravated by CS therapy. PMID- 11936022 TI - Nutritional status of renal transplant patients. AB - OBJECTIVE: To assess the effect of renal transplantation on the nutritional status of patients. DESIGN: Prospective descriptive study. SETTING: Renal Transplant Clinic at Tygerberg Hospital, Western Cape. SUBJECTS: Fifty-eight renal transplant patients from Tygerberg Hospital were enrolled in the study. The sample was divided into two groups of 29 patients each: group 1, less than 28 months post-transplant; and group 2, more than 28 months post-transplant. OUTCOME MEASURES: Nutritional status assessment comprised biochemical evaluation, a dietary history, anthropometric measurements and a clinical examination. RESULTS: Serum vitamin B6 levels were below normal in 56% of patients from group 1 and 59% from group 2. Vitamin B6 intake, however, was insufficient in only 14% of patients from group 1 and 10% from group 2. Serum vitamin C levels were below normal in 7% of patients from group 1 and 24% from group 2, while vitamin C intake was insufficient in 21% and 14% of patients from groups 1 and 2 respectively. Serum magnesium levels were below normal in 55% of patients from group 1, and in 28% from group 2. Serum albumin and cholesterol levels increased significantly during the post-transplant period in the total sample (P = 0.0001). There was also a significant increase in body mass index (P = 0.0001) during the post-transplant period. CONCLUSIONS: Several nutritional abnormalities were observed, which primarily reflect the side-effects of immunosuppressive therapy. The causes, consequences and treatment of the vitamin B6 and vitamin C deficiencies in renal transplant recipients need further investigation. PMID- 11936023 TI - Elim hospital--the first 100 years. Part 2. PMID- 11936024 TI - Time to decriminalise cannabis? PMID- 11936025 TI - Time to decriminalise cannabis? PMID- 11936026 TI - Craniocerebral gunshot injuries. PMID- 11936027 TI - Microbiological contamination of compressed air used in dentistry: an investigation. AB - The purpose of this preliminary investigation was twofold: 1) to examine the possibility of cross-contamination between a dental-evacuation system and the compressed air used in dental operatories and 2) to capture and identify the most common microflora in the compressed-air supply. The investigation used swab, water, and air sampling that was designed to track microorganisms from the evacuation system, through the air of the mechanical room, into the compressed air system, and back to the patient. Samples taken in the vacuum system, the air space in the mechanical room, and the compressed-air storage tank had significantly higher total concentrations of bacteria than the outside air sampled. Samples of the compressed air returning to the operatory were found to match the outside air sample in total bacteria. It was concluded that the air dryer may have played a significant role in the elimination of microorganisms from the dental compressed-air supply. PMID- 11936028 TI - Foot-and-mouth disease: a review of the virus and the symptoms. AB - Foot-and-mouth disease virus (FMDV) is the etiologic agent of foot-and-mouth disease (FMD), which is a disease of cattle, swine, and other cloven-footed animals. FMD is characterized by the formation of vesicles on the tongue, nose, muzzle, and coronary bands of infected animals. The virus has several unique characteristics that enable it to cause one of the most economically devastating diseases in today's world. The ease with which it may be transmitted by contact and aerosol, combined with its enhanced ability to initiate infections, virtually ensures that most, if not all, animals in a herd will contract FMD. The long-term survival of FMDV in infected animals' tissues and organs, especially when refrigerated, offers an opportunity for its national and international transmission through the food chain. Multiple serotypes and numerous subtypes reduce the effectiveness and reliability of vaccines. The possible development of carriers in vaccinated animals and those that have recovered from FMD provides additional potential sources of new outbreaks. These features create a disease that can have a major economic impact on farmers and entire nations. PMID- 11936029 TI - Funding on-site wastewater system improvements with linked-deposit loans. PMID- 11936030 TI - Case not closed--issues of police authority, municipal smoking bans, and first amendment rights. PMID- 11936031 TI - Arguing the truth before Judge Judy. PMID- 11936032 TI - When experience challenges received wisdom: the case of drinking water. PMID- 11936033 TI - A school waterborne outbreak involving both Shigella sonnei and Entamoeba histolytica. AB - In an outbreak of gastroenteritis affecting 730 students, Shigella sonnei and Entamoeba histolytica were isolated from the stool specimens of patients. Environmental investigations revealed the source of infection to be contamination of underground well water by sewage from a toilet. The outbreak ended with the closure of the well water supply. To avoid such problems, institutions and other groups that maintain their own wells, including schools and summer camps, need to be vigilant about maintenance and check for potential contamination. PMID- 11936034 TI - Iowa Dental Association Annual Session. PMID- 11936035 TI - IDA looks at direct reimbursement (DR). PMID- 11936036 TI - The past 150 years in Iowa since 1846. PMID- 11936037 TI - Medicaid reform. No progress at state or federal level. PMID- 11936038 TI - Mixed feelings. PMID- 11936039 TI - Technical quality of a minor injuries unit. PMID- 11936040 TI - Dealing with verbal abuse. PMID- 11936042 TI - Alcohol health education programme for schools. PMID- 11936043 TI - New A&E strategy to integrate care will boost nursing, claims minister. PMID- 11936041 TI - Methaemoglobinaemia and poisoning. PMID- 11936044 TI - Practice through reflection. PMID- 11936045 TI - Faculty of emergency nursing. PMID- 11936046 TI - The varying standards of mobile medical team equipment held by London hospitals. PMID- 11936047 TI - Lead poisoning. PMID- 11936048 TI - The student as named nurse. A new model of learning. PMID- 11936049 TI - Attitudes to patients who present with suicidal behaviour. PMID- 11936051 TI - Retrospective analysis of the prevalence of asymptomatic cerebral aneurysm in 4518 patients undergoing magnetic resonance angiography--when does cerebral aneurysm develop? AB - The natural history of cerebral aneurysms was investigated by measuring the prevalence of incidentally found unruptured aneurysms in the general population and evaluating the characteristics including risk factors. 'De novo' formation of aneurysm was also demographically estimated. The prevalence of incidental aneurysm was evaluated among 4518 patients who underwent magnetic resonance (MR) angiography for various reasons in a neurosurgical institute. Double the number of patients were randomly selected from the remaining patients without aneurysm as the Control group so that sex and age group were matched to the Aneurysm group. 127 patients (2.8%) had diagnoses of aneurysm. The prevalence of asymptomatic aneurysm among middle-aged and elderly patients was predominant in women and increased with age in both sexes. Patients with aneurysms had significantly more hypertension and family history of subarachnoid hemorrhage compared to the controls. The prevalence was markedly increased in the 8th decade in men and the 7th decade in women, and new aneurysms seemed to develop predominantly around these decades. Cerebral aneurysms become detectable on MR angiography in the middle or later decades, and women tend to develop aneurysm earlier than men. Hypertension and family history of subarachnoid hemorrhage are probably risk factors for the development of aneurysm. PMID- 11936050 TI - Great expectations. PMID- 11936052 TI - Clinical analysis of 16 patients with brachial plexus injury. AB - Brachial plexus injury is very rare in neurosurgical practice, so many neurosurgeons have never experienced this problem in Japan. This study describes a clinical analysis of 16 patients aged 5 to 62 years (mean 32.9 years) who presented at our institution with brachial plexus injuries. Nine patients presented with paralysis and seven with paresis. Head injury was the most common associated injury in eight of 16 patients. Six patients were managed conservatively. All patients with C8-T1 paresis spontaneously recovered to a useful level. Surgery was performed in 10 patients: six neurolysis, two neurotization, and three nerve grafting procedures. All six patients who underwent neurolysis of the brachial plexus attained useful recovery. Four of five patients achieved useful recovery after nerve repair. Nerve grafting achieved a better outcome than neurotization in this study. The difference of outcome was attributed to the graft length. The management of brachial plexus injury is a great challenge, but surgical outcome can be improved if the optimal repair procedure is selected for brachial plexus injury. PMID- 11936053 TI - Blade crossing of a pure titanium clip applied to a cerebral aneurysm--case report. AB - Titanium aneurysm clips are superior to cobalt clips in radiological features, biocompatibility, and hypoallergenicity, but the mechanical characteristics are less advantageous. A 56-year-old man was referred to our department due to dizziness and slurred speech. Cerebral angiography unexpectedly disclosed an aneurysm of the right middle cerebral artery. The patient underwent right frontotemporal craniotomy and neck clipping of the aneurysm with a commercially pure titanium clip. After application of the clip, the clip blades crossed and nearly sheared the wall of the aneurysm. The mechanical characteristics of titanium aneurysm clips require careful consideration because of possible clip malfunction. PMID- 11936054 TI - Male prolactinoma with pituitary apoplexy concomitant with intracerebral hemorrhage--case report. AB - A 56-year-old male presented with a prolactin-secreting pituitary adenoma who suffered pituitary apoplexy associated with intracerebral hemorrhage of the caudate head and putamen. The centers of both the intratumoral hematoma and intracerebral hemorrhage appeared as hyperintense on T1-weighted and T2-weighted magnetic resonance imaging, indicating the subacute stage and probably similar timing of onset. The tumor was treated surgically 6 weeks after the onset. The causative factor for the apoplexy remains unclear. PMID- 11936055 TI - Giant meningioma of the cranial vertex--case report. AB - A 55-year-old man was admitted with a swelling on the top of his head progressively growing for 2 years. The swelling had grown to a huge size with the appearance of a double head. A huge meningioma with predominantly extracranial growth and invasion into the superior sagittal sinus was subtotally excised. The excised tumor mass weighed 1380 g. The residual tumor was treated by irradiation. No recurrence was seen at one-year follow-up examination. PMID- 11936056 TI - Intracranial germinoma with syncytiotrophoblastic giant cells in the cerebellopontine angle--case report. AB - A 23-year-old man presented with a 3-week history of left hearing disturbance and left facial nerve paresis. T1-weighted magnetic resonance (MR) imaging showed an iso-intense mass in the left cerebellopontine angle (CPA), with homogeneous enhancement with gadolinium-diethylenetriaminepenta-acetic acid. T2-weighted MR imaging showed the mass as heterogeneously iso- to hyperintense. Gross total removal of the tumor was achieved. Histological examination revealed that the tumor was a germinoma with syncytiotrophoblastic giant cells. Whole central nervous system irradiation with cisplatin-etoposide chemotherapy was performed postoperatively. He has been in good condition with no sign of recurrence for 7 years. Intracranial germ cell tumors in the CPA are very rare. Total surgical removal followed by irradiation and chemotherapy will provide a good outcome. PMID- 11936057 TI - Intradiploic arachnoid cyst identified by diffusion-weighted magnetic resonance imaging--case report. AB - A 72-year-old woman presented with an intradiploic arachnoid cyst in the occipital intradiploic space which was found incidentally by magnetic resonance (MR) imaging. Computed tomography revealed a widened diploic space and thinning of the inner and outer tables of the occipital bone. The cyst appeared as isointense to the cerebrospinal fluid on both T1- and T2-weighted images. The differential diagnosis of intradiploic epidermoid cyst could be excluded because the lesion was low intensity on diffusion-weighted MR images. Arachnoid cyst is a benign lesion, so exploratory surgery should be avoided unless the cyst is symptomatic. Diffusion-weighted MR imaging is an effective modality to distinguish diploic epidermoid cysts from arachnoid cysts. PMID- 11936058 TI - Symptomatic spinal extramedullary mass lesion secondary to chronic overdrainage of ventricular fluid--case report. AB - A 69-year-old man presented with progressive nuchal pain and spastic gait 2 years after undergoing ventriculoperitoneal (VP) shunting for a pineal astrocytoma with obstructive hydrocephalus. The neurological manifestations were compatible with radiculomyelopathy caused by an upper cervical lesion. Magnetic resonance imaging showed an enhanced extramedullary mass lesion tightly constricting the upper cervical spinal cord. The pressure of the shunt system was 150 mmH2O, and lumbar puncture revealed normal cerebrospinal fluid (CSF) pressure of 170 mmH2O. After removal of the shunt system, the clinical symptoms and neuroradiological findings markedly improved. This symptomatic spinal mass lesion was thought to be formed secondary to chronic depletion of ventricular CSF through the VP shunt. PMID- 11936060 TI - Flare up of tuberculous abscess following stereotactic aspiration--case report. AB - A 16-year-old non-immunocompromised and otherwise healthy patient presented with a tuberculous brain abscess. The abscess was aspirated stereotactically. Following the aspiration the disease process flared up and multiple daughter abscess cavities were discovered. The daughter abscesses were removed via craniotomy. Stereotactic aspiration requires care to prevent such flare up of the disease. PMID- 11936059 TI - Brain abscess following transorbital penetrating injury due to bamboo fragments- case report. AB - A 56-year-old female presented with transorbital penetrating injury caused by bamboo fragments, which resulted in brain abscess 2 weeks after the injury. Initial computed tomography (CT) of the head did not reveal the foreign bodies. However, follow-up CT demonstrated a well-defined hyperdense abnormality of 1.0 cm length in the left orbit and brain abscess in the left temporal lobe. The lesion corresponding to the hyperdense abnormality on CT appeared isointense on T1-weighted magnetic resonance (MR) imaging and hypointense on T2-weighted MR imaging. The bamboo fragments were surgically removed, and aspiration and continuous drainage were performed for the brain abscess. The postoperative course was uneventful and the patient was transferred to a local hospital with minor neurological deficits. Bamboo foreign bodies may show changes in properties on CT and MR imaging in the subacute stage. Careful radiological examination and follow-up monitoring are required for the correct diagnosis and treatment of such injuries. PMID- 11936061 TI - Protect dissent. PMID- 11936062 TI - How people with HIV/AIDS manage and assess their use of complementary therapies: a qualitative analysis. AB - The objective of this article is to provide a qualitative analysis of the practical concerns that people with HIV/AIDS have with regard to their use of complementary therapies. In-depth semistructured interviews were conducted with a diverse range of people with HIV/AIDS (N = 46). An inductive grounded approach was used to collect and analyze the data. There were five central concerns: (a) selecting which therapies to use, (b) judging which therapies work, (c) combining Western medicine with complementary therapies, (d) assessing the safety of complementary therapies, and (e) dealing with the barriers to the use of complementary therapies. A better understanding of the practical dimensions of complementary therapy use highlights the treatment and care issues that people with HIV/AIDS face and offers insights into the role that nurses might play in addressing some of these issues. PMID- 11936063 TI - Effectiveness of a home-based exercise intervention for HIV-infected adults: a randomized trial. AB - The authors conducted a randomized controlled trial to assess the impact of a 15 week (20 minutes three times per week) home-based aerobic exercise intervention versus usual care on the physical endurance, immune status, and self-reported health status of 99 HIV-infected adults. In the exercise group, there was no improvement in physical endurance or health-related quality of life (HRQOL), except in the Medical Outcomes Study-HIV Health Survey Overall Health subscale (difference = 12.1, 95% confidence interval = 2.0-22.2, p = .02). Although physical endurance levels were maintained at baseline levels in the intervention group and declined in the control group, differences between the groups were small and not significant. There were also no significant changes in CD4+ T lymphocyte counts. Exercise appears to be safe in HIV-infected patients. Improvements in physical endurance and HRQOL might result if the exercise protocol is longer or progressive. Further research is needed to establish guidelines for exercise in patients on highly active antiretroviral therapy. PMID- 11936064 TI - Incidence and correlates of physical violence among HIV-infected women at risk for pregnancy in the southeastern United States. AB - To identify the incidence and correlates of physical and sexual violence among HIV-infected women at risk for pregnancy, a cross-sectional examination was conducted within a longitudinal study of reproductive decision making. Participants consisted of 275 HIV-infected women 17 to 49 years of age (mean = 30.1 years). Women were predominantly African American (87%) and single (82%), with annual incomes of $10,000 or less (66%). Overall, 68% of the women reported experiencing lifetime physical and/or sexual violence. Before becoming HIV infected, 65% of the women reported having been physically or sexually abused. After HIV diagnosis, 33% of the women reported experiencing physical or sexual abuse. Women reporting greater violence were more likely to disclose their HIV seropositive status to their sex partner. Using logistic regression, greater intent to get pregnant (odds ratio [OR] = 0.933), decreased present life satisfaction (OR = 1.048), having three or more children (OR = 0.474), and history of drug use (OR = 0.794) significantly distinguished between women who reported physical and/or sexual violence and those who did not. PMID- 11936065 TI - Predicting intentions and condom use among Latino college students. AB - To understand the factors that predict Latinos' risk-associated behavior, this study examined whether favorable beliefs about condoms and select moderator variables are related to stronger intentions to use condoms and more reported condom use at most recent vaginal and anal intercourse among Latino college students. Latino students who attended a public university in the northeast United States were mailed a paper-and-pencil questionnaire. Results from completed questionnaires (n = 199) indicated that students who perceived greater partner approval of condom use, perceived greater peer approval of condom use, and expressed confidence in their ability to control themselves reported stronger intentions to used condoms. Perceived partner approval and impulse control was related to self-reported condom use at most recent vaginal intercourse, impulse control was related to self-reported condom use at most recent vaginal intercourse, and impulse control was related to self-reported condom use at most recent anal intercourse. These relations did not significantly differ by Hispanic ethnicity, religiosity, gender, or age. Results suggest that normative beliefs and control beliefs regarding impulse control may be especially salient in both predicting and changing condom use behavior and should be incorporated into nursing assessments and tailored interventions. PMID- 11936066 TI - Quality of care measurement in nursing home AIDS care: a pilot study. AB - A pilot study was conducted to identify which, if any, demographic or quality indicators differentiate HIV-positive patients from other long-term care patients. This study used the Minimum Data Set files for all New York state nursing homes submitted in 1997. Chi-square tests were used to assess difference in proportions of demographics and quality indicators between HIV-positive and HIV-negative patients. The HIV-positive patients tended to be between 40 and 59 years of age and male and were more likely to be Black or Hispanic. HIV-positive patients had a significantly higher prevalence of diagnosis or symptoms of depression without any treatment compared to HIV-negative patients. HIV-positive patients had significantly higher prevalence of weight loss, antipsychotic use, antianxiety/hypnotic use, and incontinence of bladder and bowel compared to HIV negative patients. This study paves the way for the development of a more appropriate quality indicator system tailored to the AIDS population and allows facilities to make necessary improvements in the quality of care offered to this vulnerable population. PMID- 11936067 TI - Reliability in testing: clinical and research applications. PMID- 11936068 TI - To tell or not to tell: the ethics of disclosure in pediatric AIDS via vertical transmission. PMID- 11936069 TI - University of California, Los Angeles scientists discover stress accelerates AIDS progression, undermines the effect of anti-HIV drugs. PMID- 11936070 TI - Atazanavir demonstrates antiviral efficacy and a favorable lipid profile at 48 weeks. PMID- 11936071 TI - Two protease inhibitor switch studies show longer virologic response in patients switching to a regimen containing Sustiva. PMID- 11936072 TI - [Gustatory sensor beyond the tongue]. PMID- 11936073 TI - [Physiopathology of wheezing in children and its management]. PMID- 11936074 TI - Bibliography. Current world literature. Transplantation. PMID- 11936075 TI - Bibliography. Current world literature. Lung and mediastinum. PMID- 11936076 TI - Bibliography. Current world literature. Melanoma and other skin neoplasms. PMID- 11936077 TI - Uterine cancer incidence in the world. PMID- 11936078 TI - Activation of Helicobacter pylori CagA by tyrosine phosphorylation is essential for dephosphorylation of host cell proteins in gastric epithelial cells. AB - Helicobacter pylori type I strains harbour the cag pathogenicity island (cag PAI), a 37 kb sequence,which encodes the components of a type IV secretion system. CagA, the first identified effector protein of the cag-PAI, is translocated into eukaryotic cells and tyrosine phosphorylated (CagAP-tyr) by a host cell tyrosine kinase. Translocation of CagA induces the dephosphorylation of a set of phosphorylated host cell proteins of unknown identity. CagA proteins of independent H. pylori strains vary in sequence and thus in the number and composition of putative tyrosine phosphorylation motifs (TPMs). The CagA protein of H. pylori strain J99 (CagAJ99) does not carry any of three putative tyrosine phosphorylation motifs (TPM-A, TPM-B or TPM-C) predicted by the MOTIF algorithm in CagA proteins. CagA,n is not tyrosine phosphorylated and is inactive in the dephosphorylation of host cell proteins. By site-specific mutagenesis,we introduced a TPM-C into CagA,. by replacing a single lysine with a tyrosine. This slight modification resulted in tyrosine phosphorylation of CagAJ99 and host cell protein dephosphorylation. In contrast, the removal of the indigenous TPM-C from CagAP12 did not abolish its tyrosine phosphorylation, suggesting that further phosphorylated sites are present in CagAP12. By generation of hybrid CagA proteins, a phosphorylation of the most N-terminal TPM-A could be excluded. Our data suggest that tyrosine phosphorylation at TPM-C is sufficient, but not exclusive,to activate translocated CagA. Activated CagAPtr might either convert into a phosphatase itself or activate a cellular phosphatase to dephosphorylate cellular phosphoproteins and modulate cellular signalling cascades of the host. PMID- 11936080 TI - Can the degree of retrograde diastolic flow in abnormal umbilical artery flow velocity waveforms predict pregnancy outcome? AB - OBJECTIVE: Reverse end-diastolic flow is the most pathological type of the umbilical artery flow velocity waveform. We aimed to investigate whether additional prognostic information can be obtained from umbilical artery waveforms in cases with reverse end-diastolic flow. SUBJECTS AND METHODS: Umbilical artery Doppler velocity waveforms from 44 fetuses with reverse end-diastolic flow were analyzed and the following parameters measured: the highest amplitude and the area below the maximum velocity curve of forward and reverse flow (A, B and C, D, respectively) and the duration of forward and reverse flow (Tc and Td, respectively). Ratios A/B, C/D and Tc/Td were calculated. The cut-off values for A/B, C/D and Tc/Td with the best predictive values for perinatal death were established with the help of receiver operating characteristics curves. The three curves were compared with each other. RESULTS: Of the three ratios, A/B and C/D had the best capacity to predict perinatal death. Both ratios had acceptable sensitivities, specificities and positive predictive values. In this regard, A/B and C/D were comparable. The cut-off values for A/B and C/D were 4.3 and 4.52, respectively. Survivors had I significantly higher A/B and C/D ratios than non survivors (P = 0.0001 and 0.0003, respectively). Significantly more fetuses with A/B or C/D below the established cut-off values had pulsations in the venous system (P < 0.05). In fetuses with a gestational age < =210 gestational days the survival rate was significantly higher in those with A/B or C/D above the cut-off values (P = 0.03 and 0.003, respectively). CONCLUSIONS: The A/B or C/D ratio can be used for quantification of the reverse end-diastolic flow waveforms in the umbilical artery and may offer additional information to the evaluation of fetal condition. PMID- 11936079 TI - ActP controls copper homeostasis in Rhizobium leguminosarum bv. viciae and Sinorhizobium meliloti preventing low pH-induced copper toxicity. AB - Two 'calcium-irreparable' acid-sensitive mutants were identified after mutagenizing Rhizobium leguminosarum bv. viciae and Sinorhizobium meliloti with Tn5. Each mutant contains a single copy of the transposon which, inserted within the actP gene, prevents expression of a P-type ATPase that belongs to the CPx heavy metal-transporting subfamily. Here, we show that both actP-knockout mutants show sensitivity to copper; omission of this heavy metal from low pH-buffered media restores acid tolerance to these strains. Furthermore, complementation of the mutant phenotype requires only the actPgene. An actP-gusA fusion in R. leguminosarum was transcriptionally regulated by copper in a pH-dependent manner.Downstream to actP in both organisms is the hmrR gene that encodes a heavy metal-responsive regulator (HmrR) that belongs to the merR class of regulatory genes. Insertional Inactivation of hmrR abolished transcriptional activation of actP by copper ions and increased the basal level of its expression in their absence. These observations suggest that HmrR can regulate actP transcription positively and negatively. We show that copper homeostasis is an essential mechanism for the acid tolerance of these root nodule bacteria since it prevents this heavy metal from becoming overtly toxic in acidic conditions. PMID- 11936081 TI - First-trimester umbilical cord diameter: a novel marker of fetal aneuploidy. AB - OBJECTIVE: To compare the umbilical cord diameter at 10-14 weeks of gestation of chromosomally normal and abnormal fetuses. METHODS: In a consecutive series of women, who were undergoing routine sonographic evaluation at 10-14 weeks of gestation, umbilical cord diameter and nuchal translucency were measured. Reference ranges for umbilical cord diameter according to gestational age and crown-rump length were constructed. Fetal karyotype was obtained at chorionic villus sampling, amniocentesis or at delivery in newborns with features suspicious for chromosomal abnormalities. RESULTS: During the study period, 784 patients met the inclusion criteria. Of these, a fetal or placental chromosomal abnormality was present in 17 cases. The mean umbilical cord diameter increased with gestational age (r = 0.41, P < 0.001). The proportion of fetuses with an umbilical cord diameter above the 95th centile was higher in the presence of fetal or placental chromosomal abnormalities than in normal fetuses (5/17 vs. 39/767, P < 0.01). Among fetuses with an abnormal fetal or placental karyotype, nuchal translucency was above the 95th centile for gestational age in 10 cases. When only fetal chromosomal abnormalities were considered (n = 14), the combined detection rate was 85.7%(12/14). CONCLUSIONS: Sonographic assessment of the umbilical cord in early gestation appears to identify a subset of fetuses at increased risk of chromosomal abnormalities. PMID- 11936082 TI - [The Twenty-Third Report on Survey of the Adverse Reaction to Radiopharmaceuticals (The 26th survey in 2000)]. PMID- 11936083 TI - Contact dermatitis. AB - Ted is a 38 year old service technician for a large chemical company servicing cooling towers at a car manufacturing plant. While blending a batch of chemicals, he accidentally splashed cooling tower fluid, Kathon, an isothiazolin-3 derivative, over his left forearm. He was wearing long sleeved overalls and safety glasses at the time. About 15 minutes later, Ted experienced some itchiness of his forearm and proceeded to wash the area. He did not change his uniform. Five hours later he experienced severe pain of the forearm. Inspection revealed a 5 cm dermal burn with obvious blistering over the left forearm. He sought medical treatment and was treated conservatively with routine burn dressings. The skin blistering healed after seven days. Ted returned to work undertaking full duties. When undertaking mixing of cooling tower chemicals he developed an itchy rash of the forearms that appeared to be spreading to his hand and shoulder areas. This skin irritation occurred even when there was no contact with cooling tower fluids. PMID- 11936084 TI - Pharmacist scope of practice: response to position paper. PMID- 11936085 TI - A payment paradigm for pharmaceutical care. PMID- 11936086 TI - Local atrophy following steroid injection. PMID- 11936087 TI - Erythropoietin use in a Jehovah's Witness patient. PMID- 11936088 TI - Severe rhabdomyolysis and cerivastatin-gemfibrozil combination therapy. PMID- 11936089 TI - Comment: unbound drug concentration versus unbound drug fraction. PMID- 11936090 TI - National Hormone and Peptide Program--National Institute of Diabetes and Digestive and Kidney Diseases: peptide hormones, subunits, antisera & other reagents available. PMID- 11936091 TI - [Peritoneal tuberculosis: 27 cases in the suburbs of northeastern Paris]. AB - SETTING: Peritoneal tuberculosis did not disappear from France during the 1990s. OBJECTIVE: To determine the characteristics of peritoneal tuberculosis in the north-eastern suburbs of Paris. METHOD: A retrospective study of cases diagnosed with peritoneal tuberculosis between 1990 and 1998 in five suburban hospitals in the north-east region of Paris. RESULTS: Twenty-seven cases of adult peritoneal tuberculosis were diagnosed. There were nine women and 18 men, with a mean age of 37.5 years, 88.9% of whom were foreign born. General and digestive symptoms- abdominal pain and/or ascites--were present in 96.3% of the cases. The mean delay in treatment was 30 days. Peritoneal involvement was isolated in 25.9% of cases, and associated with pulmonary tuberculosis in 40.7% or hepatic tuberculosis in 25.9%. Co-infection with HIV (human immunodeficiency virus) was present 14.8% of cases. Culture of ascites fluid, laparoscopy and/or laparotomy (n = 17), with directed biopsy, aided in the formal diagnosis of peritoneal tuberculosis in 59.2%. One relapse and one case of multiresistance were observed. The mean duration of treatment was 9 months (range 6-12 months). Three patients received treatment with corticosteroids, and 91.2% of the patients achieved cure without sequelae. CONCLUSION: Peritoneal tuberculosis is not rare in the Paris region. The diagnosis should be suspected in case with ascites and fever, and can be confirmed by laparoscopy with sampling for bacteriology and histology. The methods of treatment need to be standardised. PMID- 11936092 TI - Electron transfer of hemoglobin at electrodes modified with colloidal clay nanoparticles. AB - Nanostructured sodium montmorillonite was prepared via a colloidal chemical approach and deposited onto glassy carbon electrodes (GCE). Subsequently, hemoglobin was spontaneously adsorbed onto the clay membrane-modified electrode. The colloidal clay nanoparticles and the adsorbed protein were characterized by scanning electron microscopy (SEM) and atomic force microscopy (AFM). The electrochemical impedance behavior of the system was studied using a microlithographically fabricated interdigitated microsensor electrode (IME). The interaction of the clay nanoparticles with hemoglobin was investigated by UV-VIS spectroscopy and electrochemical methods. The heme protein adsorbed in this way displayed a well-defined electrode process and the electron transfer was confirmed to originate from its heme site. Furthermore, nitric oxide affects the hemoglobin electrochemistry. PMID- 11936093 TI - Electrochemical characterization of screen-printed carbonaceous electrodes for the determination of peroxidase activity in novel screen-printed flow-through modules. AB - A novel totally screen-printed flow-through cell for immunoanalysis is presented. It contained screen-printed carbonaceous electrodes, which allowed the determination of peroxidase activity through the electrochemical reduction of p benzoquinone. As different electrode materials differ strongly in their electrochemical properties, electrodes resulting from various screen-printable carbonaceous pastes were characterized using the hydroquinone/ p-benzoquinone redox couple. For most of the electrodes, cyclic voltammogram peak separations of between 550 and 670 mV were observed indicating only quasi-reversible electrochemical behavior. This was confirmed by variation of the peak separation with scan rate. Heterogeneous electron transfer rates of ca. 0.5 - 1 x 10(-3) cm s(-1) and electrochemical activation energies of ca. 20 kJ mol(-1) were found. These flow-through cells were not only applied to electrochemical peroxidase activity determinations but also, in combination with a separate detector, as affinity reactors. After biotinylation of screen-printed layers, streptavidin and then biotinylated peroxidase could be bound. However, as signals were only 10-20% of those obtained with a column filled with biotinylated glass beads, only the screen-printed electrochemical detector was applied to the detection of antibodies against the African Swine Fever Virus. PMID- 11936095 TI - Thick film biosensors for metabolites in undiluted whole blood and plasma samples. AB - The new electrochemical thick film biosensors from Roche Diagnostics are presented. Following considerations about the principal requirements that biosensors have to fulfil to be useful for diagnostic purposes, the basic design of these thick film biosensors is shown. In this paper, the new generation of biosensors for glucose, lactate and urea are presented, as well as data from a new biosensor for creatinine. All biosensors are designed for multiple use, at minimum 500 samples or 1 week in-use (depending on type of enzyme used), for determinations in undiluted whole blood or plasma, with extra electrodes to compensate for interferences. The sensors are integrated in a disposable cassette requiring 38 microtl sample volume. The analytical ranges of the sensors scope well with the normal and pathological concentrations of metabolites in human blood, e.g. for glucose 0.5-40.0 mmol/L. Both biosensors and interference compensating electrodes are developed to have a cycle time of 90 s maximum. Method comparison diagrams show excellent correlation of results obtained by biosensors compared to results achieved by reference methods. In addition, the possibility of urea and creatinine determinations in diluted urine is presented. PMID- 11936094 TI - Cyclic voltammetric simulation of electrochemically mediated enzyme reaction and elucidation of biosensor behaviors. AB - A cyclic voltammetric simulation that can be applied to an electrochemically mediated enzyme reaction involving any substrate and mediator concentration was developed. Concentration polarization of the substrate in the vicinity of an electrode was considered as well as mediator concentration. Reversible electrochemical reaction with one electron followed by an enzyme reaction with two electrons was modeled. The differential equations for the mediator and substrate were solved using digital simulation techniques. The calculated cyclic voltammograms showed prepeaks when there was a low substrate concentration, high mediator concentration, and high enzyme activity. The prepeak was experimentally observed in the case of an enzyme electrode co-immobilized with a redox polymer. The enzyme electrode loaded at high redox polymer and high enzyme content showed a prepeak at low substrate concentration in the cyclic voltammogram. PMID- 11936096 TI - Cloning, functional expression and kinetic characterization of pesticide selective Fab fragment variants derived by molecular evolution of variable antibody genes. AB - Fab antibody fragments were constructed by subcloning single chain Fv variable regions from the phagemid vector pCANTAB 5E into the expression vector pASK99. The vector was designed for bacterial secretion of Fab fragments and bears coding sequences for murine constant domains including the Strep-tag II at the carboxyl terminal end of the constant heavy chain domain. The cloning procedure was carried out with the scFv antibodies IPR-7, IPR-53 and IPR-23. The second and third clone originated from the molecular evolution of the s-triazine selective antibody IPR-7. The Fab fragments were expressed under the transcriptional control of the tetA promoter system. Large-scale production benefits from the anhydrotetracycline-inducible system because of the lower costs for the inducer compared to IPTG and the tightly regulated expression of the recombinant antibody fragments. The Strep-tag purification technology facilitates the isolation of Fab fragments from the E. coli periplasm. The characteristics of functionally expressed Fab fragments were determined by employing a BIAcore 2000 system. The KD of the Fab variant IPR-23 (K(D)= 1.1 2 x 10(-9) M) optimized by molecular evolution was improved by a factor of 24 compared to the Fab IPR-7 (K(D) = 2.73 x 10(-8) M), which was derived from the template scFv antibody IPR-7. The affinity alteration was also reflected in the 22-fold reduction of the IC50 values of the variants Fab IPR-7 (IC50 = 60.5 microg/L) and Fab IPR-23 (IC50=2.7 microg/L) in the corresponding atrazine ELISA. PMID- 11936097 TI - Rapid detection of neurotoxic insecticides in food using disposable acetyicholinesterase-biosensors and simple solvent extraction. AB - The extensive use of pesticides to protect agricultural crops necessitates reliable tools for the detection of residues in food and water, thus ensuring environmental protection and consumer safety. Neuroinhibitors such as organophosphates and carbamates in particular, represent a potential hazard to human health. These compounds are frequently found in food, but conventional methods of analysis are limited as they are either time consuming or not sufficiently sensitive. As a result, a rapid and sensitive biosensor test based on AChE-inhibition was developed. The disposable AChE-biosensor was directly applied in solvent extracts of food samples using isooctane as extraction solvent. A complete assay could be performed in less than 2 h. Recovery rates of 84% were obtained in tests with spiked orange juice samples. Tests in food samples with a lower water content resulted in reduced recovery rates (44% for peach pap baby food). Phosphorothionate insecticides in food could be detected after direct oxidation with N-bromosuccinimide and solvent extraction. The assay displayed a detection limit of 2 microg/kg paraoxon, which was sufficient for the monitoring of maximum residue limits in food according to EU regulations. PMID- 11936098 TI - Potentiometric phosphate-sensing system utilizing phosphate-binding protein. AB - A phosphate-detection system has been developed which uses phosphate-binding protein (PBP) from Escherichia coli. PBP was immobilized on a sheet of nitrocellulose membrane by cross-linking and the membrane potential of the immobilized PBP was measured. The response time of the system to phosphate was 5 min. The response was selective to phosphate among other anions. Under optimum conditions 0.1-1.5 mmol L(-1) phosphate can be determined with this system. PMID- 11936099 TI - Enzyme sensor array for the determination of biogenic amines in food samples. AB - An enzyme sensor array for the simultaneous determination of the three biogenic amines (histamine, tyramine and putrescine) by pattern recognition using an artificial neural network and its application to different food samples is described. A combination of a monoamine oxidase, a tyramine oxidase and a diamine oxidase (with specific activities sufficient for rapid detection) are immobilised each on a separate screen-printed thick-film electrode via transglutaminase and glutaraldehyde to compare these cross-linking reagents with regard to their suitability. To calculate the amount of a specific biogenic amine, the raw data from multichannel software were transferred to a neural network. The sensor array takes 20 min to complete (excluding statistical data analysis) with only one extraction and subsequent neutralisation step required prior to sensor measurement. The lower detection limits with the enzyme sensor were 10 mg/kg for histamine and tyramine, and 5 mg/kg for putrescine with a linear range up to 200 mg/kg for histamine and tyramine and 100 mg/kg for putrescine. The application area of the enzyme sensor array was tested from fish to meat products, sauerkraut, beer, dairy products, wine and further fermented foods and compared with the data of conventional LC analyses (mean correlation coefficient: 0.854). PMID- 11936100 TI - Biosensors and flow-through system for the determination of creatinine in hemodialysate. AB - Biosensors for the determination of creatinine have been developed and integrated into a flow-through system. The sensors are based on a screen-printed three electrode transducer with a platinum working electrode. Applying the multi-enzyme sequence of creatininase (CA), creatinase (CI) and sarcosine oxidase (SO) hydrogen peroxide has been detected amperometrically. An optimal enzyme load was found to be 4.4 U/0.28 U/0.20 U (CA/CI/SO) and 0.28 U/0.20 U (CI/SO) per electrode for the creatinine sensor and for the creatine sensor, respectively. Among a variety of polymers Nafion has shown the highest efficiency to exclude interfering substances like ascorbic acid, acetaminophen and uric acid. First determinations of creatinine in dialysate samples obtained during hemodialysis treatments have shown a good correlation to the conventional methods, the Jaffe reaction (y=0.945x+ 2.8, R=0.9882, n=9) and the enzymatic photometric method (y=0.891x+3.5, R=0.9917, n=9). PMID- 11936101 TI - Characterisation of antibodies and analytes by surface plasmon resonance for the optimisation of a competitive immunoassay based on energy transfer. AB - The determination of binding constants using surface plasmon resonance (SPR) was introduced to optimise a competitive homogeneous fluorescence energy-transfer immunoassay (ETIA) before labelling. Steroids were chosen as model for the detection of three analytes estrone, estradiol and ethinylestradiol--by taking three polyclonal antibodies (anti estrone-, anti estradiol- and anti estrogen antibodies) and the corresponding analyte derivatives used for the immunisation. The active concentration of the antibodies was determined before and after labelling. Inhibition curves were recorded using SPR for all possible combinations of analyte, antibody, and analyte derivatives. The experiments revealed that the active antibody concentration can be reduced to 30% whereas the antibody affinity is not affected by the labelling process. Limits of the use of SPR for determination of affinity constants in solution are discussed. All possible ETIA calibration for the quantification of estrone and estradiol was performed. The lower limits of detection for estrone (0.06 microg L(-1)) and estradiol (0.17 microg L(-1)) were reached with the anti-estrogen IgG and its derivative PMID- 11936102 TI - Miniature surface-plasmon resonance immunosensors--rapid and repetitive procedure. AB - Miniaturized immunosensors based on surface-plasmon resonance (SPR) have been developed by using the sensor recently developed by Texas Instruments. By using this sensor for human immunoglobulin G (IgG) rapid and repetitive measurements could be performed by alternate injection of sample and glycine-HCl buffer. A sensor for human serum albumin (HSA) could also be developed similarly. One difficulty in immunosensor development is dissociation of the strongly bound antigen-antibody complex. A "regenerable" immunosensor has also been developed for this reason. PMID- 11936103 TI - Molecular recognition by indoleacetic acid-imprinted polymers--effects of 2 hydroxyethyl methacrylate content. AB - Molecularly imprinted polymers for indole-acetic acid were prepared by co polymerizing N,N-dimethylaminoethyl methacrylate, 2-hydroxyethyl methacrylate (HEMA), and ethylene glycol dimethacrylate. The dependence of the affinity and selectivity of the imprinted polymers on HEMA content was evaluated chromatographically. The affinity was improved by increasing the HEMA content; the selectivity of the imprinted polymer was best when the HEMA content was approximately 30%, irrespective of monomer content. PMID- 11936104 TI - Determination of the turnover number of the restriction endonuclease EcoRI using evanescent wave technology. AB - Binding and catalytic activity of the type II restriction endonuclease EcoRI on immobilized DNA has been observed in real time using three different evanescent wave biosensors and two different immobilization techniques. The method gives direct access to the turnover number (kcat) without the necessity for the determination of any concentration or activity. The combination of different evanescent wave techniques gives access to the catalytic mechanism and allows the determination of the rate limiting step. PMID- 11936105 TI - Quartz-crystal sensors for biosensing and chemical analysis. AB - The principle and applications of quartz-crystal sensors based on the three basic concepts for mass, viscosity, and viscoelastic changes are reported. In the general discussion the realization of a resonant frequency-resonant resistance diagram is described in detail. As an example of application to mass sensing, gas sensing with a carbon-coated quartz crystal is reported. Determination of the blood coagulation factor is used as an example of the application to viscosity sensing. As an example of viscoelastic measurement, an ion-exchange polymer coated quartz crystal is investigated to show that viscoelasticity changes more than mass in the transport process. The possibility of developing new biosensors and chemical sensors is discussed on the basis of these results. PMID- 11936106 TI - A disposable acetylcholinesterase-based electrode biosensor to detect anatoxin a(s) in water. AB - Anatoxin-a(s) is a hazardous toxin released by cyanobacteria during bacterial blooms. A simple and fast method to detect this hazardous compound using a biosensor based on the electrochemical detection of the activity of acetylcholinesterase was developed. Among several acetylcholinesterases, electric eel enzyme was found to be the most sensitive to anatoxin-a(s) and was thus used to build disposable amperometric sensors. The system displayed a detection limit of 1 microg/L anatoxin-a(s). No unspecific effect was noticed with real water samples but spiked toxin was accurately detected. Oxime reactivation was used to discriminate between the toxin and potential insecticides present in the sample. PMID- 11936107 TI - Chemiluminescence assay for uric acid in human serum and urine using flow injection with immobilized reagents technology. AB - A novel chemiluminescence (CL) flow sensor for the determination of uric acid in human urine and serum has been developed by using controlled-reagent-release technology. The reagents involved in the chemiluminescence (CL) reaction, luminol and periodate, are immobilized on anion-exchange resin packed in a column. After injection of water, chemiluminescence generated by released luminol and periodate in alkaline media is inhibited in presence of uric acid. By measuring the decreased chemiluminescence (CL) intensity the uric acid is sensed. The decreased response is linear in the 5.0-500.0 ng mL(-1) range, with a detection limit of 1.8 ng mL(-1). The flow sensor showed remarkable operational stability and could be easily reused for over 80 h with sampling frequency of 100 h(-1). The proposed sensor was applied to the determination of uric acid in human urine and serum, and monitoring metabolic uric acid in human urine with RSD less than 3.0%. PMID- 11936108 TI - Application of cause-and-effect diagrams to the interpretation of UV-Vis spectroscopic data. AB - The application of cause-and-effect diagrams to the evaluation of thermodynamic data from UV-Vis absorption spectroscopic analysis is demonstrated. The contributions of measurement uncertainty identified from a cause-and-effect diagram are implemented into a Monte Carlo procedure based on the threshold bootstrap computer-assisted target factor analysis (TB CAT). This algorithm aims at an improvement of data comparability and accounts for non-normality, spectral, residual and parameter correlation as well as random noise in target factor analysis. The ISO Type-B measurement uncertainties are included into the process by normally distributed random numbers with specified mean values and dispersions. The TB CAT procedure is illustrated by a flow diagram and a case study of Nd(III) complexation by picolinic acid N-oxide (pic NO) in aqueous solution. Using 12 experimental spectra as input data, the single component spectra and the formation constant 1g betaML of the Nd(pic NO)2+ species are obtained together with the respective probability density distributions. The role of the cause-and-effects approach on the further development of chemical thermodynamics is discussed. PMID- 11936109 TI - Systematic optimisation of high-performance liquid chromatographic separation by varying the temperature, gradient, and stationary phase. AB - Optimisation of the separation of a synthetic drug mixture by HPLC is performed by changing both continuous variables, i.e. mobile phase composition and temperature, and categorical variables, here the stationary phase. The retention of solutes is described on the basis of a general linear model in which the different columns are modelled by indicator variables. From the solute-specific retention models the global separation optimum is evaluated on the basis of multidimensional window diagrams using relative retentions of all peak pairs as the figure-of-merit. PMID- 11936110 TI - The bromination of acetone. application to multicomponent kinetic determinations. AB - A very simple kinetic method is proposed for the resolution of mixtures of acetone and a second component. It is based on the reaction, at constant temperature, between bromine and acetone. This reaction can be regarded, under certain conditions, as pseudo-zero-order on the bromine concentration. To show the possibilities of the method, mixtures of acetone (between 2 and 20 x 10(-4) mol L(-1)) and either hydroquinone (between 0.4 and 2.2 x 10(-4) mol L(-1)) or resorcinol (between 0.1 and 10.7 x 10(-4) mol L(-1)) have been used with the concentration ratios [acetone]:[hydroquinone] ranging between 0.5 and 50 and [acetone]:[resorcinol] ranging between 2.8 and 200. No systematic errors were found to exist (90% confidence level) and random errors were mainly under 5%. The method can be extended to mixtures of acetone and a second component whose reaction with bromine is fast (phenols, aromatic amines, etc.). PMID- 11936111 TI - Time-resolved laser-induced plasma spectrometry for determination of minor elements in steelmaking process samples. AB - A pulsed Nd:YAG laser operating on the fourth (266 nm) and second (532 nm) harmonics has been used to generate plasmas on the target surface in air at atmospheric pressure. The influence of wavelength on quantitative analysis of 4 minor elements in stainless steel samples (Si, Ti, Nb and Mo) was investigated. Stainless steel samples with different elemental concentrations were prepared and analyzed by laser-induced plasma spectrometry (LIPS). The effect of laser wavelength on analytical figures of merit (calibration curves, correlation coefficients, linear dynamic ranges, analytical precision, and accuracy values) was found to be negligible when internal standardization (an Fe line) and time resolved laser-induced plasma are employed. For both wavelengths, the calibration curves presented a good linearity and an acceptable linear dynamic range in the concentration interval investigated. For the four elements studied, limits of detection lower than 150 microg g(-1) were achieved. To evaluate the influence of wavelength on precision and accuracy, a set of fifteen high-alloyed steel samples from different stages of steelmaking process have been analyzed. Finally, the long-term stability of the analytical measurements for Mo with 532 nm wavelength has been discussed. RSD values were lower than 5.3% for the elements studied. PMID- 11936112 TI - Acid digestion of geological and environmental samples using open-vessel focused microwave digestion. AB - The application of open vessel focused microwave acid digestion is described for the preparation of geological and environmental samples for analysis using inductively coupled plasma-mass spectrometry (ICP-MS). The method is compared to conventional closed-vessel high pressure methods which are limited in the use of HF to break down silicates. Open-vessel acid digestion more conveniently enables the use of HF to remove Si from geological and plant samples as volatile SiF4, as well as evaporation-to-dryness and sequential acid addition during the procedure. Rock reference materials (G-2 granite, MRG-1 gabbros, SY-2 syenite, JA-1 andesite, and JB-2 and SRM-688 basalts) and plant reference materials (BCR and IAEA lichens, peach leaves, apple leaves, Durham wheat flour, and pine needles) were digested with results comparable to conventional hotplate digestion. The microwave digestion method gave poor results for granitic samples containing refractory minerals, however fusion was the preferred method of preparation for these samples. Sample preparation time was reduced from several days, using conventional hotplate digestion method, to one hour per sample using our microwave method. PMID- 11936113 TI - Determination of 3,5,6-trichloro-2-pyridinol (TCP) in water by a continuous competitive immunoassay system based on the streptavidin-biotin interaction. AB - A competitive continuous immunoassay system for the determination of 3,5,6 trichloro-2-pyridinol (TCP), the major degradation product of the insecticide chlorpyrifos, in water is described. The immunoassay system is based on the transient retention of the specific LIB-MC2 monoclonal antibody anti-TCP as a biotinylated derivative using the streptavidin-biotin interaction. The permanent immobilization of streptavidin on controlled-pore-glass provides an adequate active support for the transient retention of the biotinylated monoclonal antibody anti-TCP. In a subsequent step, the immuno-competitive reaction between the biotinylated LIB-MC2 and the TCP/hapten-POD mixture takes place. This competitive assay relies on the determination of the biocatalytic action of peroxidase, retained in the active support, on a derivatization reaction which yields a fluorescent product. The method exhibits a determination range of 0.01 200 microg L(-1) of TCP (r2=0.9919, n=9) with a precision, expressed as RSD, lower than 4.2% and a sampling frequency of 3 h(-1). The approach has been applied to the determination of TCP in water with recoveries of 89.7-105.6%. PMID- 11936114 TI - An improved clean-up strategy for simultaneous analysis of polychlorinated dibenzo-p-dioxins (PCDD), polychlorinated dibenzofurans (PCDF), and polychlorinated biphenyls (PCB) in fatty food samples. AB - The study and extension of a simple automated clean-up method for polychlorinated dibenzo-p-dioxins (PCDD) and polychlorinated dibenzofurans (PCDF) to a broad range of polychlorinated biphenyls (PCB) is described. The isolation of seven PCDD, ten PCDF, and three coplanar PCB (cPCB) is extended to eight monoortho substituted PCB and seven so-called "marker PCB" (Aroclor 1260) for fatty food samples. This enables quantification of 35 compounds - including all congeners with a WHO toxic equivalent factor (TEF)--in a single extraction and single purification step. The chromatographic behaviour of mono-ortho PCB and marker PCB on a variety of adsorbents, including basic alumina, has been studied. Partitioning of analytes through multi-column sequences is described and correlated with their structural and electronic properties, by use of molecular modelling calculations. The fractionation process available with the Power-Prep automated clean-up system enables rapid independent analysis of the different groups of compounds. Gas chromatography with high resolution mass spectrometry (GC-HRMS) is used for the PCDD/F and cPCB fraction and quadrupole ion-storage tandem in time mass spectrometry (GC-QISTMS) for analysis of the remaining PCB. A comparison study was performed on quality-control samples and real fatty food samples to evaluate the robustness of the new strategy compared with a reference method. On the basis of this simultaneous clean-up, a rapid simplified strategy for PCDD/F and selected PCB analysis determination is proposed for fatty food samples. PMID- 11936115 TI - Speciation of organotin compounds in marine sediments by capillary column gas chromatography-atomic absorption spectrometry coupled with hydride generation. AB - A method for speciation of organotin compounds in marine sediments by solvent extraction combined with hydride generation gas chromatography-atomic absorption spectrometry has been developed. Sediment samples spiked with tributyltin and triphenyltin chlorides were homogenized in hydrochloric acid. The chlorides were extracted twice into toluene. Recoveries of the organotin compounds from the spiked sediment samples were improved by the addition of 8-quinolinol. Tributyltin and triphenyltin chlorides form ion-associates with 8-quinolinol in aqueous hydrochloric acid. The method was optimized with respect to derivatization reactions and extraction conditions. Interferences from Sn(II/IV) and additional 13 ions were investigated. Recoveries of 84-100% for tributyltin and 86-100% for triphenyltin were achieved using this method. The detection limits obtained for tributyltin and triphenyltin chlorides were 95 and 145 pg, respectively, corresponding to a relative detection limit of 95 and 145 ng kg(-1) in the sediment. PMID- 11936116 TI - Fluorimetric determination of arsanilic acid by flow-injection analysis using on line photo-oxidation. AB - A flow-injection-fluorimetric method for the determination of arsanilic acid is proposed. The assay is based on the on-line decomposition of arsanilic acid in the presence of peroxydisulfate on irradiation with UV light. The arsenate generated in the photochemical reaction was reacted with molybdate in dilute nitric acid to form arsenomolybdic acid, which oxidised thiamine to thiochrome. The thiochrome was monitored fluorimetrically at 440 nm with excitation at 375 nm. The calibration graph was linear in the range 0.10-10.8 microg mL(-1) with a correlation coefficient of 0.999. The detection limit was 0.01 microg mL(-1) and the sample throughput was 55 samples h(-1). The applicability of the method was demonstrated by determining arsanilic acid in animal foodstuffs and water. PMID- 11936117 TI - Chromatographic characterization of molecularly imprinted microspheres for the separation and determination of trimethoprim in aqueous buffers. AB - Molecularly imprinted microspheres (MIMs) against trimethoprim (TMP), prepared by aqueous microsuspension polymerization, bound strongly to TMP, by electrostatic and other non-covalent interactions. The effects of pH, kind and ionic strength (I) of buffer on capacity factors (k') have been discussed in detail. The capacity factors for TMP increased with increasing pH of both acetate and phosphate buffers. The effects of ionic strength on capacity factors were very substantial and the linear relationship between logk' and logI was described by the equation logk'=0.3162-0.4420logI with R=-0.9995. The results showed that pH 3.5 acetate buffer (0.05 mol L(-1)) containing 0.1 mol L(-1) sodium chloride and a 1:9 ratio of buffer to methanol were the optimum conditions for separation and determination of TMP. The calibration plot of peak area against concentration was linear with R=0.9979. PMID- 11936118 TI - Radiochemical neutron-activation analysis of uncertified ultra-trace rare earth elements in two biological certified reference materials. AB - Radiochemical neutron activation analysis (RNAA) has been used for the determination of eight rare earth elements (La, Ce, Nd, Sm, Eu, Tb, Yb, and Lu) in two Chinese certified reference materials (CRM), GBW 08503 (wheat powder) and GBW 09101 (human hair). These determinations are important for possible certification of the above mentioned ultra-trace elements, so far not certified. A simple one-step (REE)F3 precipitation was used. Chemical yields were determined for all relevant elements by means of tracer experiments. The two CRM were also analyzed by inductively coupled plasma-mass spectrometry (ICP-MS) to compare the merits and draw-backs of these two major trace analytical techniques for these particular elements. RNAA was proven to be a reliable technique for ultra-trace analysis, especially in the certification of some ultra-trace elements. PMID- 11936119 TI - A simplified method for the HPLC resolution of alpha-carotene and beta-carotene (trans and cis) isomers. AB - A new HPLC/DAD (Diode Array Detector) method is proposed for the identification of some carotene isomers. The operating conditions adopted permit the resolution of alpha-carotene, all-trans-beta-carotene, 9-cis-beta-carotene, 13-cis-beta carotene and 15-cis-beta-carotene. Moreover, the chromatographic conditions reported are simplified in respect of those reported up to now. The method is applied to the determination of carotenoids in a dried Dunaliella salina extract, but it could be also applied to other organic matrices such as eggs. PMID- 11936120 TI - Treatment of acute hepatitis C with interferon alfa-2b. PMID- 11936121 TI - Treatment of acute hepatitis C with interferon alfa-2b. PMID- 11936122 TI - Treatment of acute hepatitis C with interferon alfa-2b. PMID- 11936123 TI - B vitamins and restenosis after coronary angioplasty. PMID- 11936124 TI - Nephrectomy for metastatic renal-cell cancer. PMID- 11936125 TI - Nephrectomy for metastatic renal-cell cancer. PMID- 11936126 TI - Nephrectomy for metastatic renal-cell cancer. PMID- 11936127 TI - No cash to implement NICE, health authorities tell MPs. The National Institute for Clinical Excellence. PMID- 11936128 TI - Quarter of Dutch paediatric intensive care beds closed. PMID- 11936129 TI - Canada sees a bigger role for private companies in health care. PMID- 11936130 TI - King's Fund calls for NHS to be made a corporation. PMID- 11936131 TI - Polish police hold health workers over alleged murder of patients. PMID- 11936132 TI - Commentary: Do genes or environment influence development of rheumatoid arthritis? PMID- 11936133 TI - Revalidating retired doctors. Retired doctors' skills would be invaluable in a crisis. PMID- 11936134 TI - Use of interactive multimedia decision aids. Discrepancy may exist between GPs and their patients about who is making the decisions. PMID- 11936135 TI - Widespread body pain and mortality. Diagnosing fibromyalgia stops doctors from thinking. PMID- 11936136 TI - An evaluation of the Ethical, Legal and Social Implications program of the U.S. Human Genome Project. AB - The Ethical, Legal, and Social Implications (ELSI) program is an in-house evaluation program for the United States Human Genome Project. Understanding the ethical and moral implications of genetic information and technology is crucial towards ensuring the proper use of genetic data. The ELSI programs have had a positive influence in understanding problematic areas surrounding the HGP by acting as a center for discussion for many bioethicists and scientists. However, ELSI is often too passive and does not provide enough practical guidance to the public on the complex implications of the HGP. As it stands now, there is room for major improvement such as increased cooperation between ELSI and non government organizations. NGO's would serve as bridges between ELSI and the public, and a collaboration would enable in-depth probing and more comprehensive analysis of issues of public concern. PMID- 11936137 TI - The historical contingency of bioethics. AB - The principles of bioethics have been historically contingent, a product of social values, circumstances, and experience. During the early twentieth century, they rested on a doctor-knows-best autonomy that permitted physicians to perform research on human subjects with a minimum degree, if any, of informed consent. The eugenics movement of the period embraced an implicit bioethics by presuming to sterilize individuals for the sake of a larger social benefit, a practice and doctrine that helped lead to the Nazi medical experiments and death camps. After World War II, the promulgation of the Nuremberg Code failed to halt eugenic sterilization and risky human experimentation without informed consent either in civilian or military venues. However, beginning in the 1960s, these practices came under mounting critical scrutiny, partly because of the increasing attention given to individual rights. By now, it is widely understood that concern for individual rights rather than an appeal to some national good belongs at the heart of bioethics. PMID- 11936138 TI - Provision of health care: how much, if any, are we entitled to? AB - Over 45 million Americans currently lack health insurance, and this number continues to rise. In light of convincing evidence that being uninsured or underinsured produces adverse health outcomes, it is incumbent upon us to define our ethical standards in terms of providing health care. This essay considers the ethical arguments for and against providing universal health care, and considers what possible goals a policy of universal care should aim for. Ultimately, an ethical commitment to securing basic opportunity in the realms of employment, personal life, and social life requires us to provide health care to all Americans. The paper further concludes that the goal of such a plan must be a guarantee of access to basic health care, although not necessarily equal care, for all Americans. PMID- 11936139 TI - Organ procurement and mandated choice: an alternative to the existing system. AB - One of the greatest problems facing medicine today is the allocation of scarce resources. A manifestation of this problem that affects everyone is the procurement and allocation of donor organs. This paper examines the current system of donor organ procurement in the US, points to its weaknesses, and examines three proposed alternatives. It proposes the adoption of one these alternatives, the system of mandated choice, wherein all potential donors are required to make an informed and legally binding decision to donate or not. PMID- 11936140 TI - Gerwirth's ethical rationalism and abortion: a response. AB - In the preface to his seminal work, Reason and Morality (1978), Alan Gerwith writes: "The most important and difficult problem of philosophical ethics is whether a substantial moral principle can be rationally justified." After summarizing his methodology, I demonstrate that (1) Gerwith's attempt to quantify personhood is unrealistic; (2) that his position on abortion rests on the unintelligible notion of "comparable conflict" between mother and unborn; and (3) that he implicitly assumes that personhood is naturally, and not functionally, defined--thereby contradicting himself. Ultimately, I outline an alternative view of personhood, one which avoids the criticism to which Gerwith's theory is particularly susceptible--namely, that personhood is a natural component of human beings from the start, rather than a gradually acquired trait. PMID- 11936141 TI - Stem cell research: biology, ethics, and policy. AB - Human stem cell research may have many promising benefits, including giving us the ability to regenerate human tissue and organs, understand the process of early human development, and provide better models to test new drugs. However, it is very controversial and in many cases presently illegal because stem cells may be derived from human embryos or from a cloning process. This paper outlines the various techniques which may be used for deriving stem cells, the ethical problems such research presents, and suggestions for creating policy for human stem cells research. PMID- 11936142 TI - CIRSE '96 summary report. Proceedings of the Annual Meeting of CIRSE. Madeira, Funchal, September 8-12, 1996. PMID- 11936143 TI - Is there a false memory syndrome? A review of three cases. AB - The controversy over recovered memories of childhood sexual abuse (CSA) is whether such experiences can be forgotten for long periods and retrieved later in therapy or in response to cues or triggers from the environment. False memory syndrome (FMS) is caused by memories of a traumatic experience--most frequently CSA--which are objectively false, but in which the person strongly believes. Personality factors often play a role in the development of FMS. Because CSA is such a devastating experience, false accusations of sexual abuse have enormous, if not shattering, consequences for families. We present three case reports to illustrate features of the FMS. FMS should be listed for further study to establish valid criteria for making the diagnosis under the category of "factitious disorders," and a subcategory of "false memories/beliefs of abuse," with a further subdivision of "induced by therapy." The FMS controversy occurred in the context of a general moral panic about sexual abuse in the early 1980s. Psychiatrists should have a high degree of scepticism to moral panics. PMID- 11936144 TI - The public perceptions of the rewards and risks of genetic research: the Oregon story. PMID- 11936145 TI - The value of difference: nonaffiliates on IRBs provide alternative views. PMID- 11936146 TI - IRB roles and responsibilities: an outline of duties for institution, IRB, administrator, and investigator. PMID- 11936147 TI - Research intermediaries: position created to protect vulnerable people who might not have capacity to consent. PMID- 11936148 TI - How should we justify universal access to health care--freedom or determinism, justice or charity? PMID- 11936149 TI - Catholic hospitals and unions. PMID- 11936150 TI - Dying with dignity: crying for clarity. PMID- 11936151 TI - The convergence of abortion regulation in Germany and the United States: a critique of Glendon's Rights Talk thesis. PMID- 11936152 TI - The body bazaar: blood, kidneys, eggs, sperm--name a body part, there's a price on it. Think of it as a commodities market for the twenty-first century. PMID- 11936153 TI - Substance abuse in pregnancy and the child born alive. PMID- 11936154 TI - Legalising active euthanasia and physician assisted suicide. People's autonomy is not absolute. PMID- 11936155 TI - Legalising active euthanasia and physician assisted suicide. Seeking this presumed moral good is immoral. PMID- 11936157 TI - Legalising active euthanasia and physician assisted suicide. Strong evidence base must be adduced for euthanasia. PMID- 11936158 TI - Legalising active euthanasia and physician assisted suicide. Patients need medical help to live with dignity until they die naturally. PMID- 11936159 TI - Legalising active euthanasia and physician assisted suicide. Denying people voluntary euthanasia causes unnecessary suffering. PMID- 11936161 TI - National screening programme for diabetic retinopathy. Screening by retinal photography offers holistic package of diabetic care. PMID- 11936162 TI - Infections and risk factors in entrants to Irish prisons. Study in Zambia showed that robust response is needed in prisons. PMID- 11936163 TI - The politics of peanut butter. AB - Conflicting studies and special interest groups make developing policies to protect public health contentious and chaotic. How can lawmakers recognize 'sound' science and make good decisions? PMID- 11936164 TI - New challenge for states: Indian health care. AB - Devolution has given states a role in making sure Indian tribes have adequate health care. Along with these new opportunities comes greater responsibility--for both states and tribes. PMID- 11936165 TI - From laboratories to legislatures ... legislating in the genomic age is harder than policymakers ever thought. PMID- 11936166 TI - Medicaid woes returning. AB - After lying dormant for several years, Medicaid is again giving lawmakers headaches as they tackle budget problems. PMID- 11936167 TI - Professionalism and the AuD. PMID- 11936168 TI - Professionalism and the audiology student: characteristics of master's versus doctoral degree students. AB - Only scarce documentation is available concerning the professional characteristics of students within audiology training programs. In the current study, audiology students were asked to complete a questionnaire that explored some of the major issues of student professionalism. Equal numbers of questionnaires were sent to both Doctor of Audiology (AuD) and master's degree programs to determine the characteristics of students being attracted to and trained within two different courses of study. Audiology students as a whole ranked low in most aspects of professionalism when compared with other students in health care. Similarities were observed between the two student groups for the majority of questions. Statistically significant differences were calculated with regard to future employment, income, and autonomy. Master's students more commonly wished to become employees rather than employers. Significantly higher numbers of master's than AuD students expect audiology to provide them with a secondary source of income. Finally, more master's than AuD students report doubt concerning the future autonomy of audiology. PMID- 11936169 TI - Effects of training on timbre recognition and appraisal by postlingually deafened cochlear implant recipients. AB - The purpose of this study was to compare the effect of structured training on recognition and appraisal of the timbre (tone quality) of musical instruments by postlingually deafened cochlear implant recipients. Twenty-four implant users (Nucleus CI24M) were randomly assigned to a control or a training group. The control group experienced only incidental exposure to music in their usual daily routine. The training group participated in 12 weeks of training delivered via a laptop computer in which they were introduced to excerpts of musical instruments representing three frequency ranges and four instrumental families. Those implant recipients assigned to the training group showed significant improvement in timbre recognition (p < .0001) and timbre appraisal (p < .02) compared to the control group. Correlations between timbre measures and speech perception measures are discussed. PMID- 11936170 TI - Effects of stimulus rate and gender on the auditory middle latency response. AB - The effects of stimulus rate and gender on the auditory middle latency response (AMLR) waveforms were examined in 20 young adult male and female subjects. Four different repetition rates were presented to subjects (1.1/sec, 4.1/sec, 7.7/ sec, and 11.3/sec). Stimulus repetition rate had a significant effect on Pa latency, Pa amplitude, and Pb amplitude. Pa and Pb amplitudes decreased with increasing the stimulus rate, and Pa latency significantly increased with increasing the stimulus rate. No significant differences were seen on Pb latency or site of recording. Gender had a significant effect on Pa latency and Pa amplitude. Pa latencies were longer in male subjects, and Pa amplitudes were larger in female subjects. Gender did not have a significant effect on the Pb waveform. PMID- 11936171 TI - Effects of stimulus presentation level on stop consonant identification in normal hearing and hearing-impaired listeners. AB - The purpose of the present study was to determine whether varying the presentation level of stop consonant stimuli resulted in similar phonetic boundary shifts for listeners with normal and impaired hearing. Sixteen normal hearing and 16 hearing-impaired listeners categorized synthetic speech stimuli as /b/, /d/, or /g/. The onset frequency of F2 varied from 900 to 2300 Hz (100-Hz steps), and the presentation level varied from 92 to 62 dB SPL (10-dB steps) for each stimulus presentation. Hearing-impaired listeners had significantly more missing boundary values than normal-hearing listeners; however, the correlation between the number of missing boundary values and hearing sensitivity was not significant. Comparison of boundary shift with level demonstrated that hearing impaired listeners had a smaller boundary shift with increasing level than normal hearing listeners. The amount of boundary shift was not correlated with audibility. The results of the current study suggest that increasing the presentation level of a signal does not result in performance similar to that of listeners with normal hearing. PMID- 11936172 TI - Hearing loss and hearing handicap in users of recreational firearms. AB - This investigation sought to establish the prevalence of hearing loss and hearing handicap in a population of 232 recreational firearm users. Hearing handicap was calculated based on four methods using pure-tone threshold data from the American Academy of Ophthalmology and Otolaryngology, American Academy of Otolaryngology Head and Neck Surgery, National Institute of Occupational Safety and Health, and American Speech-Language and Hearing Association in addition to the self-report Hearing Handicap Inventory for Adults-Screener (HHIA-S). Subjects (45 female and 187 male) ranging in age from 13 to 77 years (mean = 40 years, SD = 15.1) completed a short questionnaire regarding demographics and shooting practices followed by pure-tone air audiometry at Occupational Safety and Health Administration test frequencies of 500 to 6000 Hz. A total of 177 who exhibited varying degrees of hearing loss also received a face-to-face administration of the HHIA-S. Audiometric and HHIA-S results revealed that both high-frequency hearing loss and hearing handicap varied significantly as functions of age and occupation. Significant gender effects were observed audiometrically but not as a function of hearing handicap. HHIA-S scores varied significantly as a function of high-frequency (1000-4000 Hz) hearing loss. Correlation coefficients between the four different pure-tone methods of calculating hearing handicap and the self reported HHIA-S were highest for pure-tone methods that do not employ 500 Hz in the calculation. PMID- 11936173 TI - Otoacoustic emissions. PMID- 11936174 TI - In vivo reflectance measurement of optical properties, blood oxygenation and motexafin lutetium uptake in canine large bowels, kidneys and prostates. AB - Motexafin lutetium (MLu) is a second-generation photosensitizer for photodynamic therapy (PDT) of cancer. We have developed and applied a diffuse optical reflectance spectrometer for in vivo measurement of MLu uptake, optical properties, haemoglobin concentration and haemoglobin oxygen saturation in normal canine large bowels, kidneys and prostates. The probe consists of a broadband fibre-optic-coupled light source and detector fibres placed at various distances from the source fibre to collect reflected light. An analysis based on the diffusion approximation of the photon transport equation was used to recover tissue optical properties from the reflectance measurements. The instrumentation and analysis methods were validated using measurements from homogeneous, highly scattering phantoms with known MLu concentrations. The same techniques were then used to estimate chromophore concentrations of normal canine large bowels, kidneys and prostates. We estimated (mean (standard deviation)) total haemoglobin concentrations of 119 (25), 340 (92) and 51 (11) microM in the large bowels, kidneys and prostates of four dogs, respectively; tissue blood oxygen saturations in these same organs were 75 (15), 76 (21) and 74 (16) per cent, respectively. Tissue MLu concentrations (mg l(-1)) were estimated from data taken 3.5 h after injection of a 2 mg kg(-1) injected dose; data from three dogs gave concentrations of 2.4 (0.4) in large bowels, 6.8 (1.3) in kidneys and 2.2 (1.1) in prostates. The reduced scattering coefficients, mu's, estimated for large bowels, kidneys and prostates at 730 nm were, respectively: 10.1 (1.3), 19.6 (4.0) and 12.7 (0.6) cm(-1). We observed significant variability in MLu uptake, tissue scattering and haemoglobin concentration between organs and even between the same organ in different dogs. This class of in situ optical property measurement may be desirable to individualize PDT drug and light delivery. PMID- 11936175 TI - Determination of the dose characteristics in the near area of a new type of 192Ir HDR afterloading source with a pinpoint ionization chamber. AB - Since the company MDS Nordion Haan GmbH introduced the new afterloading radiation unit GammaMed plus in 1998, new HDR Ir-192 sources with a diameter of 0.9 mm (formerly 1.1 mm) and a length of 4.5 mm (formerly 5.5 mm) have been used. With this equipment it is possible to treat peripheral vessels for prophylaxis of restenosis after percutaneous transluminal angioplasty (PTA). In this case a good knowledge of the dose characteristics in the near area is the prerequisite for safe application. In this study, the dose characteristics for this source type is investigated at close range by means of an ionization chamber. For the description of the dose characteristics, the radial dose distribution and the anisotropy function are used. The measurements were carried out using an ionization chamber, the so-called PinPoint chamber (Type PTW 31006, PTW Freiburg, Germany). The radial dose distribution was examined in the area of 0.26 to 12 cm. The anisotropy function was examined in the range from 0.5 to 5 cm (distance from the effective site of measurement to the source axis). The radial dose function was compared with the correction function k(as,w) (r), published in the German standard DIN 6809 part 2 (1991 Beuth Verlag, Berlin, Germany). In the area up to 4 cm (distance source-axis to measuring point) the deviation maximized up to 2%. The deviation climbs with greater distance from the source with results around 3%. The anisotropy function was compared with that used in the treatment planning system Abacus 3.0 of the same manufacturer. In the close range between 0.5 and 1 cm, there were some major variations (up to 10%) from the anisotropy function used in the planning system. The fault reduces itself with greater distance to under 2%. In view of the results presented in this paper further measurements of the anisotropy function in the close range from 0 to 1.5 cm from the source axis should be made. It can be seen that the function used in the planning system supplies results that are too high in comparison with the results of other authors. This concerns primarily the area of very small and very big angles. PMID- 11936176 TI - Comparison between a conventional treatment energy and 50 MV photons for the treatment of lung tumours. AB - Radiation therapy in the thoracic region is difficult due to the presence of many dose-limiting structures and the large density differences that affect the dose distribution. Conventional irradiation techniques use low-energy photon beams to avoid build-up effects superficially in the tumour and increased lateral scattering of the beams. For deep-seated tumours higher beam energies could have lung-sparing properties that would enable dose escalation. A comparison was made for a conventional low photon energy (6 MV) and 50 MV photons for the treatment of a lung tumour. A representative patient geometry was selected, consisting of a small tumour semi-enclosed in lung tissue. Treatment plans were designed using a commercial 3D-pencil beam treatment planning system. The treatment beams designed in the TPS were simulated with the Monte Carlo code EGS4/BEAM and the dose distribution in the phantom created from the patients CT-data was calculated using MCDOSE with identical beam geometry for both energies. The intrinsic difference between the two photon energies implies a sparing effect of lung that can be utilized for dose escalation. For a treatment with two beams the mean total dose to the tumour could be increased by 5.3% for 50 MV, corresponding to 3.2 Gy for a prescription dose of 60 Gy, with the same complication probability for the treated lung as for 6 MV. In conclusion, high-energy beams have qualities that can be taken advantage of for irradiation of lung tumours. Optimum solutions would probably require the use of both high- and low-energy beams. PMID- 11936177 TI - An optimization algorithm that incorporates IMRT delivery constraints. AB - An intensity-modulated beam optimization algorithm is presented which incorporates the delivery constraints into the optimization cycle. The optimization algorithm is based on the quasi-Newton method of iteratively solving minimization problems. The developed algorithm iteratively corrects the incident, pencil-beam-like, fluence to incorporate the delivery constraints. In the present study, the goal of the optimization algorithm is to achieve the best deliverable radiotherapy plan, subject to the constraints of the delivery technique described by a leaf-sequencing algorithm being applied concurrently. In general, if they are applied after, rather than during, the optimization cycle, the delivery constraints associated with the IMRT technique can produce local variations up to 6% in the 'optimized' dose (i.e., distribution without applied constraints) and reduce the degree of conformity, of the dose, to the PTV region. The optimization method has been applied to three IMRT delivery techniques: dynamic multileaf (DMLC), multiple-static-field (MSF) and slice-by-slice tomotherapy (NOMOS MIMiC). The beam profiles were generated for a prostate tumour with organs at risk being the rectum, bladder and femoral heads. The optimization method described was shown to generate optimum and deliverable IMRT plans for these three delivery techniques. In the case of the DMLC and MSF the optimization converged within 3-5 iterations to a mean PTV dose of 69.60 +/- 1.34 Gy and 69.71 +/- 1.34 Gy, respectively, while for NOMOS MIMiC approximately 10 iterations were needed to obtain 69.68 +/- 1.55 Gy. In addition to this, the IMRT optimization also yielded optimum fluence profiles when clustering was performed concurrently with the leaf sequencer. An optimum between 8 and 15 clusters of equal fluence 'intensity' was shown to establish the best compromise between the number of fluence levels and the PTV dose coverage. PMID- 11936178 TI - A Monte Carlo simulation model of mammographic imaging with x-ray sources of finite dimensions. AB - A simulation model of mammographic x-ray sources with finite size has been developed. The model is based on Monte Carlo methods and it takes into account the electron penetration inside the anode, the anode geometry and material, as well as the resulting heel effect and the spectral and spatial distribution of x rays. This x-ray source simulation model has been embedded into an earlier developed simulation package of a mammography unit. The main outputs of this model are Monte Carlo generated images that correspond to the irradiation of properly designed phantoms. In this way it is possible to make studies of the influence of x-ray source characteristics on MTF. This paper presents the development of the mammographic x-ray source model, accompanied by a set of simulation studies concerning the influence of magnification effects as well as that of the x-ray spatial and spectral distribution on the mammographic spatial resolution for a certain magnification factor (m = 1.4). The validity level of the model, as well as its limitations and perspectives, rise through comparisons with experimental and theoretical data. PMID- 11936179 TI - Reconstruction of biologically equivalent dose distribution on CT-image from measured physical dose distribution of therapeutic beam in water phantom. AB - From the standpoint of quality assurance in radiotherapy, it is very important to compare the dose distributions realized by an irradiation system with the distribution planned by a treatment planning system. To compare the two dose distributions, it is necessary to convert the dose distributions on CT images to distributions in a water phantom or convert the measured dose distributions to distributions on CT images. Especially in heavy-ion radiotherapy, it is reasonable to show the biologically equivalent dose distribution on the CT images. We developed tools for the visualization and comparison of these distributions in order to check the therapeutic beam for each patient at the National Institute of Radiological Sciences (NIRS). To estimate the distribution in a patient, the dose is derived from the measurement by mapping it on a CT image. Fitting the depth-dose curve to the calculated SOBP curve also gives biologically equivalent dose distributions in the case of a carbon beam. Once calculated, dose distribution information can be easily handled to make a comparison with the planned distribution and display it on a grey-scale CT-image. Quantitative comparisons of dose distributions can be made with anatomical information, which also gives a verification of the irradiation system in a very straightforward way. PMID- 11936180 TI - Measurement and correction of the effects of lag on contrast-detail test results in fluoroscopy. AB - Persistence of the video signal between TV frames, an effect also known as image lag, can lead to anomalously good contrast-detail test results for fluoroscopy systems. In this practical paper, a simple method is described which quantifies lag in fluoroscopy systems and corrects for its effect on threshold contrast. A digital framestore was used to acquire temporally contiguous fluoroscopy images. Correlation of the variance between an initial base TV frame and successive later frames was then measured via the correlation coefficient. Plotted against time, this function defines a time constant which characterizes the rate at which the initial variance pattern is replaced by incoming quantum noise. A survey of seven fluoroscopy units incorporating vacuum TV camera tubes found a mean time constant of 0.06 s. The relative change in contrast-detail performance was then measured as a function of applied digital frame averaging for two separate fluoroscopy units. A time constant was found for each frame averaging mode using the correlation of variance between frames. These measurements were used to derive a function which corrects contrast-detail results obtained for a unit with a measured nominal time constant to the typical vacuum camera tube time constant of 0.06 s. The correction is shown to significantly reduce the spread of contrast detail results obtained over a range of temporal filtration settings. PMID- 11936181 TI - Iterative solution of dense linear systems arising from the electrostatic integral equation in MEG. AB - We study the iterative solution of dense linear systems that arise from boundary element discretizations of the electrostatic integral equation in magnetoencephalography (MEG). We show that modern iterative methods can be used to decrease the total computation time by avoiding the time-consuming computation of the LU decomposition of the coefficient matrix. More importantly, the modern iterative methods make it possible to avoid the explicit formation of the coefficient matrix which is needed when a large number of unknowns are used. To study the convergence of iterative solvers we examine the eigenvalue distributions of the coefficient matrices. For the sphere we show how the eigenvalues of the integral operator are approximated by the eigenvalues of the coefficient matrix when the collocation and Galerkin methods are used as discretization methods. The collocation method approximates the eigenvalues of the integral operator directly. The Galerkin method produces a coefficient matrix that needs to be preconditioned in order to maintain optimal convergence speed. With the ILU(0) preconditioner iterative methods converge fast and independent of the number of discretization points for both the collocation and Galerkin approaches. The preconditioner has no significant effect on the total computational time. PMID- 11936182 TI - In vivo validation of the design rules of the coronary arteries and their application in the assessment of diffuse disease. AB - The conventional rationale that uses per cent diameter reduction to assess diffuse coronary artery disease is not appropriate because no normal reference segments exist. In a recent publication, we have proposed a theoretical model based on physical principles that relate the various morphological and haemodynamic parameters (cross-sectional area, length, volume and flow) of the normal coronary arterial tree. The model was validated using haemodynamic simulations based on detailed morphological data of the pig coronary arterial tree. This paper extends the model validation to in vivo swine studies. Coronary arteriography was performed in five swine (15-18 kg body weight) after power injection of contrast material into the coronary artery. Coronary arterial length was obtained using a 3D reconstruction technique. The arterial volume, cross sectional area and blood flow were measured using videodensitometry. The proposed relationships between these quantities were validated. Furthermore, a sensitivity analysis was demonstrated based on a simulation of diffuse coronary artery disease (approximately 40% reduction in cross-sectional area). The results of a sensitivity analysis based on a simulation of diffuse coronary artery disease suggest that the relationships between arterial volume, cross-sectional area, blood flow and the distal arterial length can be utilized to quantify moderate levels of diffuse coronary artery disease. PMID- 11936183 TI - The effects of paramagnetic contrast agents on metabolite protons in aqueous solution. AB - The longitudinal (R1) and transverse (R2) relaxivities of the clinically used contrast agents Gd(DTPA)2-, Gd(DOTA)- and Gd(DTPA-BMA) have been determined in mixed aqueous metabolite solutions for choline, creatine and N-acetylaspartate. Measurements were performed at 1.5 T using a STEAM sequence on 25 mM metabolite solutions at pH = 7.4 and 22 degrees C. The data showed that for all the contrast agents and metabolites, R1 approximately = R2. The largest range of relaxivity values was found for Gd(DTPA)2-, where R2 = 6.8 +/- 0.3 mM(-1) s(-1) for choline and 1.5 +/- 0.4 mM(-1) s(-1) for N-acetylaspartate. Variation in relaxivity values was attributed primarily to differences between the charges of the paramagnetic agent and metabolite. The maximum potential influence of the contrast agents on in vivo metabolite signals was calculated using the measured relaxivities. PMID- 11936184 TI - Consecutive projections onto convex sets. AB - In this note we describe and evaluate the performance of a novel approach to information recovery that involves consecutive projection onto convex sets (POCS). The method is applied to a time series of medical image data and the results are compared to images reconstructed using the standard POCS reconstruction method. The consecutive POCS method converges in a desired step wise manner producing reconstructed images of superior quality compared to the standard scheme and can speed up the reconstruction process. The proposed method is of value for many finite sampling imaging problems including, in particular, fast-scan magnetic resonance imaging applications. PMID- 11936185 TI - Electronic portal imaging devices: a review and historical perspective of contemporary technologies and research. AB - A review of electronic portal imaging devices (EPIDs) used in external beam, megavoltage radiation therapy is presented. The review consists of a brief introduction to the definition, role and clinical significance of portal imaging, along with a discussion of radiotherapy film systems and the motivations for EPIDs. This is followed by a summary of the challenges and constraints inherent to portal imaging along with a concise, historical review of the technologies that have been explored and developed. The paper then examines, in greater depth, the two first-generation technologies that have found widespread clinical use starting from the late 1980s. This is followed by a broad overview of the physics, operation, properties and advantages of active matrix, flat-panel, megavoltage imagers, presently being commercially introduced to clinical environments or expected to be introduced in the future. Finally, a survey of contemporary research efforts focused on improving portal imaging performance by addressing various weaknesses in existing commercial systems is presented. PMID- 11936186 TI - Analysis of cell-free human alpha1 integrin with a monoclonal antibody to the I domain: detection in ocular fluid and function as an adhesion substrate. AB - The alpha1 beta1 integrin, an inserted (1) domain containing collagen receptor, is expressed in the cell surface membrane of normal and malignant cells, and may play a role in their migration through tissues or in metastatic spread. Here we report that a functional anti-human alpha1beta1 integrin monoclonal antibody (mAb) (1B3.1) directly and specifically binds plastic bound recombinant human alpha1 I-domain protein containing the collagen binding site. Detection was diminished by acidification of the I-domain protein but was enhanced by increasing concentrations of Mg2+ cation. Furthermore, we detected binding of the mAb to proteins from the ocular fluids of 6 patients, with the highest concentration, corresponding to 22.1 ng/ml of I-domain, found in a sample from the eye of a patient with metastatic lung adenocarcinoma. Interestingly, we found that both SKNSH neuroblastoma cells and virally transformed human T cells adhered specifically to plastic wells coated with either immobilized collagen IV or alpha1 I-domain. MAb I B3.1 inhibited adhesion to collagen IV but not to immobilized I-domain. These results suggest a novel function for cell free alpha1 I-domain as a substrate for cellular adhesion, which may have relevance in tumor spread in vivo. PMID- 11936187 TI - Fibronectin promotes calcium signaling by interferon-gamma in human neutrophils via G-protein and sphingosine kinase-dependent mechanisms. AB - A common intracellular signal activating polymorphonuclear leukocytes (PMN) in inflammation is a change in cytosolic calcium concentration. Previously, we have shown that interferon-gamma (IFN-gamma) induces transient calcium signals in PMN, but only after intracellular calcium store depletion. Using a digital imaging system, we show that adhesion of PMN is critical for IFN-gamma-induced calcium signals, and with PMN attached to the optimal coating, the calcium signals are evoked even in presence of extracellular calcium, that is, non-depleted calcium stores. Adhesion to fibronectin, pure or extracted from plasma by gelatin, improved the IFN-gamma responses compared with serum, plasma, or vitronectin coats. In accordance with previous observations, IFN-gamma-induced calcium signals in fibronectin adherent cells were totally abolished by the G-protein inhibitor pertussis toxin and were also inhibited by the sphingosine kinase inhibitors dimethylsphingosine (DMS) and N-acetylsphingosine (N-Ac-Sp). PMN contact with fibronectin alone, measured in cells sedimenting onto a fibronectin coated surface or by addition of fibronectin to glass-adherent cells, evoked transient calcium signals. However, PMN in suspension did not respond to the addition of fibronectin or arginine-glycine-aspartate (RGD). The fibronectin induced calcium signals were also clearly depressed by pertussis toxin and by the sphingosine kinase inhibitors DMS, dihydrosphingosine (DHS), and N-Ac-Sp. When the product of sphingosine kinase activity, sphingosine 1-phosphate (S1-P), was added to the cells, similar calcium signals were induced, which were dependent on a pertussis toxin-sensitive G-protein activity. Finally, addition of S1-P to the cells prior to stimulation with IFN-gamma partly mimicked the priming effect of fibronectin. In conclusion, fibronectin contact evokes by itself a calcium signal in PMN and further promotes calcium signaling by IFN-gamma. We suggest that fibronectin might activate sphingosine kinase, and that the sphingosine 1 phosphate thereby generated induces a calcium signal via a G-protein-dependent mechanism. Apparently, sphingosine kinase activity is also involved in IFN-gamma induced calcium signals. PMID- 11936188 TI - Correlated motion and oscillation of neighboring cells in vitro. AB - It has long been realized that fibroblastic and epithelial cells establish recognizable patterns in tissue culture. This behavior implies that neighboring cells interact with one another to produce organized populations. Interaction between cells that are separated by many intervening cells is also possible and is demonstrated here using a special configuration of a biosensor referred to as electric cell-substrate impedance sensing (ECIS). Normally the electrical impedance of a single electrode covered with a confluent cell layer is measured, and the morphological changes of the cells are reflected in the impedance. In this case the cells are cultured on two closely spaced electrodes whose impedances are measured independently as a function of time, and communication between the cell populations is revealed as a correlation between these two time series. We also report for the first time another striking manifestation of dynamic cell interaction, where confluent layers of Madin-Darby canine kidney cells (MDCK) on a single electrode are observed to oscillate in synchrony with a period of approximately 2.5 h. PMID- 11936189 TI - Prominent beta-5 gene expression in the cardiovascular system and in the cartilaginous primordiae of the skeleton during mouse development. AB - The alpha v beta (alpha(v)beta5) heterodimer has been implicated in many biological functions, including angiogenesis. We report the beta5 gene expression pattern in embryonic and foetal mouse tissues as determined by Northern blotting and in situ hybridization. During the earliest stages, beta5 mRNA is widespread in the mesoderm. During later developmental stages, it remains mostly confined to tissues of mesodermal origin, although probable inductive effects trigger shifts of beta5 gene expression from some mesenchymatous to epithelial structures. This was observed in the teeth, skin, kidneys, and gut. Of physiological importance is the beta5 labeling in the developing cardiovascular and respiratory systems and cartilages. Furthermore, early beta5 gene expression was observed within the intra- and extraembryonic sites of hematopoiesis. This suggests a major role for beta5 in the hematopoietic and angiogenic stem cells and thus in the development of the vascular system. Later, the beta5 gene was expressed in endothelial cells of the vessels developing both by angiogenesis and vasculogenesis in the lung, heart, and kidneys. Moreover, the beta5 hybridization signal was detected in developing cartilages but not in ossified or ossifying bones. beta5-Integrin is a key integrin involved in angiogenesis, vasculogenesis, hematopoiesis, and bone formation. PMID- 11936190 TI - Anatomical correlations of intrinsic axon repair after partial optic nerve crush in rats. AB - About 15% of retinal ganglion cells survive diffuse axonal injury of the optic nerve in adult rats. Following initial blindness, discrimination of visual stimuli in behavioral tests recovers within three weeks. To investigate the mechanisms promoting this functional recovery the axonal transport and the neurofilaments were studied. Intraocularly applied MiniRuby is transported until the place of crush and accumulated in enlarged axon terminals. Three weeks after lesion the anterograde transport of MiniRuby recovers distal to the place of crush. At the same point in time the retrograde transport of surviving retinal ganglion cells is restored which was visualized by horseradish peroxidase injected into the superior colliculus. The heavy neurofilament was stained immunohistochemically and analyzed statistically up to three weeks after optic nerve crush. The stained filaments in the axon fibers of retinal ganglion cells appear wavelike and/or fragmented up to day 8, but first signs of heavy neurofilament restitution in the fibers of the optic nerve are seen at day 12 after axonal injury. Because these results cannot be explained by longlasting axon regeneration, the present results provide convincing evidence for intrinsic axon repair soon after diffuse axonal injury that correlates in time with recovery of vision. PMID- 11936191 TI - Comparative quantitative study of the intrinsic cardiac ganglia and neurons in the rat, guinea pig, dog and human as revealed by histochemical staining for acetylcholinesterase. AB - This study was conducted to determine the overall number of intrinsic neurons distributed through-out the entire heart, in which most neurons are located inside of intramural ganglia and are hidden to observers. For this reason, we attempted to ascertain: (1) how the number of neurons located inside of intrinsic cardiac ganglion is related to its area, and (2) whether this relationship is dependent on age and species of animals. Hearts of rats, guinea pigs, dogs and humans were used to examine intramural ganglia stained histochemically for acetylcholinesterase (AChE). The number and parameters of neurons located inside of 104 ganglia were estimated in serial sections. Although the revealed intrinsic cardiac ganglia varied extremely in shape and size, two different types were identified: the globular and plain ones. In the plain ganglia, perikarya of side by side situated neurons were always intensely stained for AChE and, being clearly discernible, they could be reliably counted in any plain ganglia on total heart preparations using a contact microscope. Contrarily, neuron somata in the globular ganglia were densely packed above one another and their perikarya were almost indiscernible for the observer. Counting of neurons located inside of globular ganglia was possible in serial sections only. The largest cardiac ganglia were revealed in dogs, in which some globular ganglia containing up to 2000 neurons occupied more than 1 mm2. In spite of evident species-dependent differences with respect to frequency of large ganglia, the majority of intrinsic cardiac ganglia both in humans and animals were comparatively small, involved approximately 100-200 nerve cells and occupied an area ranging from 0.01 to 0.17 mm2. Overall, the number of neurons located inside of globular ganglion was related to its area (correlation coefficient = 0.82). However, the correlation coefficients between the globular ganglion area and its neuron number were unequal in different species (0.92 in guinea pig; 0.80 in dog; 0.72 in human; and 0.44 in rat) as well as dependent on (1) ganglion size (0.8 for ganglia equal to or larger than 0.17 mm2 and 0.6 for ganglia smaller than 0.17 mm2) and (2) age of specimens (respectively, 0.98 for juvenile and 0.87 for adult dogs; 0.71 for infants and 0.54 for aged human). In all examined animals and humans, the mean measurements of neuron perikarya were similar (on average, 23 microm in width, 32 microm in length, and 615 microm2 in area) and differences between them were statistically insignificant. However, neuron perikarya of adult dogs and aged humans were significantly larger than those revealed in the juvenile dogs and infants, respectively. Based on the data of this study, we concluded that the number of intrinsic cardiac neurons may be approximated in the total heart preparation via counting and measuring of intramural ganglia, contours of which are well-discernible following a histochemical reaction for AChE. PMID- 11936192 TI - Expression of neural cell adhesion molecule (NCAM) in the peripheral nerve fibers demonstrated by postembedding immunogold method on ultrathin sections. A preliminary study. AB - The neural cell adhesion molecule, NCAM, is thought to be necessary for the interaction between neurons, axons and glial cells, axons and target structures, and for myelination. The NCAM localization in the peripheral nerves has been less well studied. The data have shown significant differences. In this study, the distribution of the NCAM immunoreactivity in the sciatic nerves of 10- and 15-day old Wistar rats was examined on ultrathin sections by the immunogold postembedding method. The sections were immunotested with a polyclonal antibody (Santa Cruz Biotechnology) that recognizes rat NCAM. The antibody was visualized with donkey anti-goat IgG, conjugated to 6 nm colloidal gold. NCAM immuno-like activity was found on axoplasmic profiles of myelinated and nonmyelinated axons, at non-assembled myelin lamellae, and at axo-glial junctions of the paranodes. The compact myelin and the Schwann cell cytoplasm were immunonegative. The presence of some NCAM immunoactivity on the level of the noncompact myelin membranes, deprived of the major myelin protein Po, might be associated with the mechanism of their maintenance. PMID- 11936193 TI - The effect of estrogens and dietary calcium deficiency on the extracellular matrix of articular cartilage in Gottingen miniature pigs. AB - Clinical observations have suggested that estrogens are involved in the pathogenesis of postmenopausal osteoarthritis (OA). However, positive and negative associations between the incidence of OA and serum estrogen concentrations have been reported. In contrast to this, osteoporosis is regarded as a disease with a strong estrogen-dependent component. Moreover, there is an interaction between estrogen and calcium deficiency: calcium supplementation potentiates the effect of estrogen therapy. The present study was designed to investigate how estrogen deficiency affects the articular cartilage depending on calcium supply. The distribution of different types of glycosaminoglycans and collagens can be used as an indicator for extracellular matrix changes induced by estrogen deficiency. Different levels of dietary calcium were therefore fed to intact and ovariectomized Gottingen miniature pigs for one year before articular cartilage was harvested. The histochemical staining for heavy sulfated glycosaminoglycans in the extracellular matrix of ovariectomized miniature pigs, especially of those fed with a low calcium diet, was stronger in comparison to intact animals. In intact animals type II-collagen was immunodetected in all zones of unmineralized and mineralized articular cartilage, while immunostaining for this protein was negative to weak in the deep radiated fiber zone of ovariectomized minipigs. These results suggest that the synthesis of heavy sulfated glycosaminoglycans and immunohistochemically detectable type II-collagen is possibly influenced by estrogen deficiency. In conclusion, under estrogen deficiency, the extracellular matrix of articular cartilage underwent similar changes to those observed in physiologically aging cartilage where keratan sulfate is increased as a heavy sulfated glycosaminoglycan. PMID- 11936194 TI - Carmine-picroindigocarmine: an alternative multiple staining method. AB - Carmine-picroindigocarmine, a multiple staining method, was developed at the beginning of the previous century by Professor Harry Kull at the University of Tartu. The stain, combining copper carmine, picric acid and indigocarmine gives bright and colourful results. Nuclear structures are stained in red, cytoplasm in varying shades from yellow to green, collagen fibres in blue, the matrix of hyaline cartilage in greyish-blue, muscle tissues from brownish-red to brownish green, erythrocytes in yellow. Squamous epithelia are stained in red with horney layers in dark red, nail plate and hairs are stained in bright yellow. The carmine-picroindigocarmine staining is stable, which allows for the combining of additional dyes without interfering with the main colouring. The combination of carmine-picroindigocarmine staining with resorcin-fuchsine in principle maintained the colouring of Kull's original method with additional staining of elastic fibres in violet colour that clearly differentiated them from blue stained collagen fibres. The described multiple staining gives an original colourful and aesthetic result, providing an alternative to other multiple staining methods. PMID- 11936195 TI - An improved method for beta-galactosidase activity detection on muscle tissue. A light and electron microscopic study. AB - In the present study we describe a method for the histochemical demonstration of bacterial beta-D-galactosidase activity on skeletal muscle tissue processed for light and transmission electron microscopy. Hence allowing this enzyme to be accurately detected, bacterial beta-galactosidase expression was studied in transgenic mouse where the enzyme, with the nuclear localization signal (nlacZ), is under the transcriptional control of the striated muscle-specific promoter MLC3F. The chromogenic substrate, 5-bromo-3-indolyl-beta-D-galactopyranoside (Bluo-Gal), was used both to recognize labelled myofibers, and beta-gal positive organelles inside single myofibers. Moreover, because the preservation of enzyme is highly dependent on tissue fixation, we developed a suitable fixation solution allowing good preservation of both tissue and enzymatic activity. This was achieved by briefly fixing tissue (3 hours) in glutaraldehyde (2.5%) and paraformaldehyde (1%) in combination. This method should be taken into consideration when studying the gene therapy of muscle diseases because it is sensitive, inexpensive and not time consuming. PMID- 11936196 TI - Morphometrically observable aging changes in the human tongue. AB - Like the outside parts of the nose and the ear but unlike most other organs, the tongue continues to grow at advanced age. Therefore, internal morphological aging processes must also proceed in a specific way. We used sectioned specimens of the radix linguae from 111 humans. The specimens were fixed in 4% formalin solution, embedded in paraffin, then cut at a thickness of 5 microm and stained with hematoxylin-eosin. The structures of interest were measured with an automatic image analyzing system (SIS, Munster, Germany). The statistical analysis package SPSS was used for correlation and regression analysis and testing of the resulting coefficients. The mean cross-sectional area of the muscle fibers increases sharply during youth, but remains at a high level into old age. After the age of seventy, it increases again. With muscles of the locomotor apparatus, this same parameter decreases after the fifth decade. We have demonstrated the process of adipose tissue formation in muscles and serous glands. Gender differences in the aging process could be clearly shown. Numerous oncocytes were visible in ducts from elder persons. PMID- 11936197 TI - Differences of morphometrical parameters in hind limb muscle fibres between ovarectomized and sexually intact female dogs. AB - Slow and fast twitch fibres of the Mm. tibialis cranialis, semitendinosus and sartorius of seven sexually intact and seven ovarectomized female beagles were histochemically and morphometrically analysed. Along with type I and type IIA fibres, another main type II fibre (IIS), which seems to be peculiar to the dog, was found in the Mm. semitendinosus and tibialis cranialis. Type I fibers comprised 26% and type II fibres 74% of all recorded muscle fibres in the M. tibialis cranialis, 29% (type I) and 71% (type II) in the M. semitendinosus and 51% (type I) and 49% (type II) in the M. sartorius, respectively. The average single profile area and the corresponding mean diameter of fibre types I and II in the investigated hind limb muscles were generally larger in ovarectomized than in sexually intact animals. This was more evident in type II than in type I fibres. However only the type II fibres of the M. tibialis cranialis and sartorius exhibited a statistically significant increase in diameter (p < 0.01 and p < 0.05, respectively). Accordingly, the mean density (number of fibres/mm2) of both fibre types in the hind limb muscles of spayed dogs was generally reduced. Again, this reduction attained statistical relevance in the type I and II fibres of the tibialis cranialis. In addition, the fibre densities of type I in the semitendinosus and type II in the sartorius muscles were also significantly reduced in ovarectomized dogs. In conclusion, ovarectomized beagles showed a generally increased mean diameter of the investigated type I and II hind limb muscle fibres and a concomitant decreased average fibre density of the respective types when compared to sexually intact animals. PMID- 11936198 TI - Collecting and paramuscular venules in glandular mucosa of rat stomach. AB - Blood from the rat gastric mucosa is drained by collecting venules running from the subepithelial layer towards the lamina muscularis mucosae. Details of their structure were studied in translucent, flat strips of the glandular stomach, in thick sections of glandular mucosa cleared in mineral oil and in semi-thin plastic sections. The number and dimensions of collecting venule outlets revealed in flat strips of gastric mucosa increased after administration of atropine and papaverine and intravital ligation of the portal vein in comparison with that of intact animals or animals with intravitally ligated portal vein but without administration of relaxing agents. In hyperemic mucosa short venules running parallel to the lamina muscularis mucosae (paramuscular venules) and draining collecting venules were distinctly visible. Saccular outlets equipped with triangular protrusions usually intervened between these vessels, probably directing blood flow. Collecting venules were straight, curved, extended or two armed. Furthermore, numerous collecting venules contained circumscribed dilatations (sacculi) connected with the lumen of the collecting venule. Connection of paramuscular and submucosal veins occurred within the muscularis mucosae. Thus, contraction of the muscularis mucosae might control the outflow of venous blood from the gastric mucosa. Conceivably, alternate contraction and relaxation of muscularis mucosae could cause expansion and collapse of collecting venules which, in turn, would facilitate the movement of glandular content to the surface of the stomach and/or movement of interstitial fluid between cells. PMID- 11936199 TI - Quantification of glands and fat in breast tissue: an experimental determination. AB - In breast surgery and anatomy, the composition of the breast itself could vary from patient to patient due to respective proportions of glandular and fatty tissue. There is no easy and reliable way to predict these proportions. The purpose of this study was to evaluate the respective proportions of glands and fat in breasts. By analysing mastectomy specimens we were able to quantify the amounts of glandular and fatty tissue in the resected breasts. Twenty one breasts were studied following mastectomy for pure in situ carcinomas. The total volume and weight of the specimens were recorded and completed by specific densities of glandular and fatty tissue for each breast. This group was submitted to a calculation of proportions of glands and fat. The percentage of fat volume in the total breast volume varied from 7 to 56% and the percentage of fat weight in the total breast weight varied from 3.6 to 37.6%. This great variability in the respective proportions of fat and glands in the evaluated specimens was not significantly correlated to age and body mass index. PMID- 11936200 TI - The postglenoid tubercle: prevalence and growth. AB - AIM: To quantify the prevalence and growth of the postglenoid tubercle in a skull sample and in children. MATERIAL: a) ninety skulls ranging in age from between 2 years and adulthood, b) sixtyfour corrected lateral tomograms of left and right temporomandibular joints of 32 boys and girls. Their age range was between 9 and 11 years. METHOD: Impressions of the temporal fossae of the skull material were taken with a silicone type impression material, using a face-bow for leveling the base of the impression parallel to the FH plane. Each impression was divided into two halves along a paramedian plane from the tip of the postglenoid tubercle through the middle of the articular eminence and the surface was photocopied to a 200% scale. Height was measured with an electronic caliper. The presence or absence of a postglenoid tubercle was established on the corrected tomograms. FINDINGS: Seventy-nine percent of the skulls had a postglenoid tubercle. It steadily enlarged and reached almost its final dimension by the age of 13 years. On corrected tomograms, 66% of the children showed a postglenoid tubercle. CONCLUSIONS: a) the postglenoid tubercle exists in a high percentage of human temporomandibular joints b) growth is almost completed by the age of 13, and c) there exists a right-left symmetry. PMID- 11936201 TI - Impact of innervation and exercise on muscle regeneration in neovascularized muscle grafts in rats. AB - The direction of known staged process of regeneration of free muscle grafts was inverted in our experimental rat model from a centripetal to a centrifugal by central implantation of blood vessels into isolated free muscle grafts. The effects of innervation, reinnervation and exercise on muscle fiber regeneration were analyzed at various intervals from 4 to 90 days by morphological and morphometric methods. Reinnervation occurred as well in grafts with the motor nerve left intact as it did in grafts with a severed and reimplanted nerve. Reinnervation proved to be prerequisite for a lasting muscle regeneration. Denervated muscle grafts even after neovascularization underwent irreversible fibrosis. A positive effect of exercise on the early states (30 days) of muscle regeneration was revealed by morphometrical analysis. In the long term (90 days) fiber diameter assimilated in all groups. The animal model mimics a clinical situation of flap prefabrication demonstrating the relationship of functional tissue regeneration and neovascularization. It can be transferred into the acute clinical situation as well as in tissue engineering. PMID- 11936202 TI - The human mandible in lateral view: elliptical fourier descriptors of the outline and their morphological analysis. AB - An original procedure based on new developments in elliptical Fourier methods associated with image analysis technics was applied to 117 human mandibles in lateral view (69 males, 48 females) in order to analyze interindividual variability and sexual dimorphism. Original parameters, called elliptical Fourier descriptors, allowed for the quantification of the shape of an outline irrespective of its morphological complexity, and provided a precise individual characterization. The use of step by step reconstructions with an increasing number of harmonics allowed for demonstration of the morphological contributions of the elliptical Fourier descriptors, and relationships to precise anatomical features were established. The first harmonics (1st to 8th) described the general mandibular shape, and the later harmonics (8th to 14th) characterized accessory and finer morphological features. The proportion of individuals presenting significant sexual dimorphism was 97.1% in males and 91.7% in females, and after size normalization 84.1% in males and 81.2% in females. Even after removal of the size component, these results in regard to mandibular dimorphism remained better than those found by classical approaches; this could be due to the use of elliptical Fourier analysis which takes into account all the information of the mandibular outline. Compared to the size component, the shape component seemed to be more important in the sexual dimorphism of the human mandible than previously recognized. The present approach opens interesting anthropological and clinical perspectives for the characterization of mandibular morphology. PMID- 11936203 TI - Personality autobiographies. Introduction. PMID- 11936204 TI - Exploring possible ethnic differences and bias in the Rorschach Comprehensive System. AB - In this study I conducted a series of analyses to explore potential ethnic bias in Rorschach Comprehensive System variables using a consecutive series of 432 patients evaluated in 1 setting. The simple association between 188 scores and ethnicity revealed no significant findings after matching on several salient demographic variables. Next, 17 analyses of convergent validity found no evidence for differential validity and no evidence for slope bias. For 13 analyses, there was also no evidence for intercept bias. However, with 4 variables predicting psychotic disorders, regression lines favored minorities and worked against European Americans. These findings are the opposite of what should be seen if the Rorschach was biased against minorities and most likely emerged for statistical reasons related to unmeasured confounds. Finally, principal components analyses revealed no evidence of ethnic bias in the Rorschach's internal structure. Although these findings need to be replicated, the available data support using the Comprehensive System across ethnic groups. PMID- 11936205 TI - Assessment training in clinical psychology doctoral programs: what should we teach? What do we teach? AB - All clinical psychology doctoral programs accredited by the American Psychological Association provide training in psychological assessment. However, what the programs teach and how they teach it vary widely. So, also, do beliefs about what should be taught. In this study, program descriptive materials and course syllabi from 84 programs were analyzed. Findings highlight commonalities in basic course content and supervised practice in administering, scoring, and interpreting assessment instruments as well as differences in coverage of psychometric and other assessment-related topics and in the extent to which lectures, labs, and practica are integrated. PMID- 11936206 TI - Comparability of MMPI-2 scales and profiles over time. AB - This study investigated Minnesota Multiphasic Personality Inventory-2 (MMPI-2; Butcher, Dahlstrom, Graham, Tellegen, & Kaemmer, 1989) scale and profile comparablilty for MMPI-2 profiles completed on 2 separate occasions by mental health patients receiving treatment at a Veterans Affairs Medical Center (n = 114). Patients were predominantly men (96.5%), with an average age of 44.08 and an average of 12.39 years of education at the time of initial testing. MMPI-2 tests were completed on 2 separate occasions as a routine part of treatment with a mean interval between test administrations of 688 days. Findings were analyzed for the complete sample and for 3 subsamples with different test-retest intervals. MMPI-2 scale test-retest correlation coefficients for the entire sample ranged from .48 to .69 for the Basic scales, .49 to .80 for the Supplementary scales, and .56 to .78 for the Content scales with scale high-point agreement = 38.60%, high 2-point agreement = 16.67%, and high 3-point agreement = 19.30%. High-point agreement for subsets of participants with well-defined high points, 2-points and 3-points was 41.07%, 27.50%, and 25.93% respectively. Pearson r correlation coefficients for T scores across the Basic scales for pairs of profiles averaged .78, suggesting similarity of profile shape across testing occasions. MMPI-2 profiles were also examined in relation to Skinner and Jackson's 3 modal MMPI profile types. PMID- 11936207 TI - Taxometric analysis of impression management and self-deception in college student and personnel evaluation settings. AB - The typology of impression management (IM), a deliberate attempt to create a positive social image, and self-deceptive positivity (SDP), an overly positive bias in self-description, was examined using taxometric procedures with Minnesota Multiphasic Personality Inventory-2 (MMPI-2; Butcher, Dahlstrom, Graham, Tellegen, & Kaemmer, 1989) underreporting scales and Balanced Inventory of Desirable Responding (Paulhus, 1988) impression management and SDP scales in a sample of college students. MMPI-2 underreporting scales were then examined in a personnel evaluation sample. IM was found to be taxonic in both student and personnel settings. Given support for the IM taxon, taxometric procedures allow the estimation of the base rate of IM and the classification accuracy of MMPI-2 IM scales in the absence of a separate criterion. Using taxometric procedures, the mean base-rate estimates were .16 and .25 for student and personnel settings, respectively. Overall classification rates ranged from .80 to .94 for MMPI-2 IM scales in the personnel setting. PMID- 11936208 TI - Bimodal score distributions and the Myers-Briggs Type Indicator: fact or artifact? AB - We examined Myers-Briggs Type Indicator (MBTI) score distributions computed using item response theory (IRT) to assess the generalizability of earlier bimodality reports that have been cited in support of the "type" versus "trait" view of personality. Using the BILOG IRT program to score a sample of approximately 12,000 individuals who participated in leadership development programs, theta score distributions for the 4 dimensions of the MBTI computed using 10 (the BILOG default) versus 50 quadrature points were compared. Results indicated that past reports of bimodality were artifacts caused by BILOG's default use of a small number of quadrature points; when larger numbers of points were used, score distributions became strongly center-weighted. Although our findings are not supportive of the "type"-based hypothesis, the extremely high correlations between theta scores (rs > .996) suggest that no practical differences would be expected as a function of the number-of-quadrature-points decision. PMID- 11936209 TI - Comment on "Effects of acculturation on the MMPI-2 scores of Asian American students". PMID- 11936210 TI - Faking psychopathy? An examination of response styles with antisocial youth. AB - Psychopaths frequently con and manipulate others in an attempt to achieve their own objectives. In the current literature, correlational research has generally found that psychopathy has (a) an inverse relation with social desirability and (b) a positive relation with malingering. Although instructive, these correlational data do not address whether the assessment of psychopathy is vulnerable to specific response styles. This study examined 2 response styles among adolescent offenders in the context of pending adjudication: social desirability and social nonconformity. On 3 measures of psychopathy (i.e., Psychopathy Checklist: Youth Version, Psychopathy Screening Device, and Self Report of Psychopathy-Second Edition), a simulation design was employed with a realistic scenario, incentives for successful deception, and appropriate manipulation checks. Results indicated moderate to large effect sizes for social desirability and large effect sizes for social nonconformity. The implications of these results for the clinical assessment of psychopathy are discussed. PMID- 11936211 TI - Speak, memory, or goodbye, Columbus. AB - This article describes the vicissitudes in the career of a psychologist who entered graduate school almost immediately after the end of World War II at a time when the standards and definitions of appropriate training in clinical psychology were still being determined. Looking back at the developments in this career, a pattern of interests, unseen at the time, can now be detected. From investigating the influence of the subject-examiner relationship in the testing situation to the experimental study of psychoanalytic propositions, first using the Rorschach test and later by means of subliminal stimulation procedures, one central theme was the uncovering of disguises and hidden motives. A second major theme, much more explicit than the first, was the primacy of data over speculation. PMID- 11936212 TI - A process dissociation approach to objective-projective test score interrelationships. AB - Even when self-report and projective measures of a given trait or motive both predict theoretically related features of behavior, scores on the 2 tests correlate modestly with each other. This article describes a process dissociation framework for personality assessment, derived from research on implicit memory and learning, which can resolve these ostensibly conflicting results. Research on interpersonal dependency is used to illustrate 3 key steps in the process dissociation approach: (a) converging behavioral predictions, (b) modest test score intercorrelations, and (c) delineation of variables that differentially affect self-report and projective test scores. Implications of the process dissociation framework for personality assessment and test development are discussed. PMID- 11936213 TI - The predictive capacity of the MMPI-2 and PAI validity scales and indexes to detect coached and uncoached feigning. AB - The objective of this study was to examine the relative effectiveness of the Minnesota Multiphasic Personality Inventory-2 (MMPI-2) and the Personality Assessment Inventory (PAI) validity scales and indexes to detect malingering. Research participants were either informed (coached) or not informed (uncoached) about the presence and operating characteristics of the validity scales and instructed to fake bad on both the MMPI-2 and PAI. The validity scale and index scores produced by these research participants were then compared to those scores from a bona fide sample of psychiatric patients (n = 75). Coaching had no effect on the ability of the research participants to feign more successfully than those participants who received no coaching. For the MMPI-2, the Psychopathology F scale, or F(p), proved to be the best at distinguishing psychiatric patients from research participants instructed to malinger, although the other F scales (i.e., F and Fb) were also effective. For the PAI, the Rogers Discriminant Function index (RDF) was clearly superior to the other PAI fake-bad validity indicators; neither the Negative Impression Management scale nor Malingering Index were effective at detecting malingered profiles in this study. Overall, RDF proved to be marginally superior to F and F(p) in distinguishing MMPI-2 and PAI protocols produced by research participants asked to malinger and psychiatric patients. Both the RDF and the F and F(p) scales, however, were able to increase the predictive capability of one another. PMID- 11936214 TI - Contributions of impulsivity and callousness in the assessment of adjudicated male adolescents: a prospective study. AB - This study examined whether delinquent adolescents with low scores on personality scales related to callousness and impulsivity would show less antisocial behavior and better psychosocial functioning compared to those elevated on 1 or both scales. A group of 162 White and African American male adolescents, recently adjudicated in a southern youth court system, were assessed at baseline and at 18 month follow-up. Dependent variables included measures of antisocial and prosocial behaviors, days detained in juvenile detention, and symptoms of psychopathology. As hypothesized, adolescents low on both callousness and impulsivity spent fewer days detained and reported less antisocial behavior and fewer symptoms of psychopathology compared to those high on I or both scales. Finally, African American participants were detained for more days, but did not report significantly more antisocial behaviors or symptoms of psychopathology compared to the White adolescents. The results suggest that being low on callousness and impulsivity may serve a protective role for future delinquent behavior for both African American and White adolescents. PMID- 11936215 TI - A proposed methodology of cancer risk assessment modeling using biomarkers. AB - A methodology for cancer risk assessment modeling was developed using a biomarker of DNA adduct, exposure dose, and tumor response. DNA adducts in the blood and lung were measured after single or multiple administration of [3H]benzo[a]pyrene (1 x BaP) in ICR mice. Making the assumption that DNA adducts are formed in a dose-dependent manner as observed in 1 x BaP treatment, kinetics patterns of DNA adducts were predicted at two other hypothetical BaP doses (2 x BaP, 1/2 x BaP) for single and continuous BaP treatments because the difference between the simulated and the experimental kinetic responses only amounted to 5.49-5.86% in terms of the integrated area under the curve. Correlations between the formation of DNA adducts and exposure doses or between blood DNA and lung DNA adducts were determined to be linear. The dose-response relationship between biomarker and exposure dose was further incorporated into a dose-tumor response equation, obtained from 2-yr bioassay, to predict cancer risk. The interrelationships between exposure dose, biomarker, and tumor response allowed the prediction of cancer risk in animals, once the information on biomarker levels was obtained. Moreover, this methodology could be further applied to human cancer risk assessment after appropriate safety factors were employed. PMID- 11936216 TI - Influence of polymorphism of GSTM1 gene on association between glycophorin a mutant frequency and urinary PAH metabolites in incineration workers. AB - The association between urinary 1-hydroxypyrene glucuronide (1-OHPG) levels, as an internal measure of polycyclic aromatic hydrocarbon (PAH) exposure, and glycophorin A (GPA) mutation frequency, as an early biologic effect indicator, was determined to establish whether genetic polymorphisms of glutathione S transferase (GST) isoforms GSTM1 and GSTT1 play a role. Eighty-one workers including 38 employees directly involved in incinerating industry wastes were recruited from a company located in South Korea. Urinary 1-OHPG levels were measured by synchronous fluorescence spectroscopy after immunoaffinity purification using monoclonal antibody 8E11. Erythrocyte GPA variant frequency (NN or NO) was assessed in MN heterozygotes with a flow cytometric assay. The GSTM1 and GSTT1 genotypes were assessed by a multiplex polymerase chain reaction (PCR) method. Urinary 1-OHPG levels were higher in workers handling industrial wastes than in those with presumed lower exposure to PAHs. An increase was seen in GPA variant frequency levels with increase in urinary 1-OHPG levels. When this association was evaluated by GSTM1 genotype status, the association between GPA mutation and urinary 1-OHPG levels was stronger in individuals with GSTM1 genotype. These results suggest that the association between urinary 1-OHPG and GPA mutation might be modulated by the GSTM1 genotype. PMID- 11936217 TI - Detection of chromosome-specific aneusomy and translocation by benzene metabolites in human lymphocytes using fluorescence in situ hybridization with DNA probes for chromosomes 5, 7, 8, and 21. AB - Benzene is a widespread human carcinogen, inducing leukemia and hematotoxicity. Exposure of human lymphocytes to benzene metabolites has been shown to cause genetic damage, including aneusomy and chromosome aberrations. In order to detect the specific chromosomal changes in chromosomes 5, 7, 8, and 21 induced by benzene metabolites, 1,2,4-benzenetriol (BT), hydroquinone (HQ), and trans,trans muconic acid (t,t-MA), fluorescence in situ hybridization (FISH) procedure in the metaphase spread of human lymphocytes was employed. Treatment with BT, HQ and tt MA resulted in the induction of monosomy 5, 7, 8, and 21 in human lymphocytes in a concentration-dependent manner. All of these metabolites also induced trisomy 5, 7, 8, and 21, but no correlation between frequencies of trisomy and concentration was found. Translocations between chromosome 8 and another unidentified chromosome [t(8:?)] and between chromosome 21 and another unidentified chromosome [t(21:?)] were found. However, translocation between chromosome 8 and 21 [t(8:2 1)] was not found. Results indicate that the benzene metabolites BT, HQ and t,t-MA induce chromosome-specific numerical and structural aberrations, and the fluorescence in situ hybridization (FISH) approach may be a useful and powerful technique for detection of aneuploidy. PMID- 11936218 TI - Effects of flavonoids isolated from Scutellariae radix on cytochrome P-450 activities in human liver microsomes. AB - A series of flavonoids isolated from Scutellariae radix were evaluated for their effects on cytochrome P-450 (CYP) activities in human liver microsomes. All flavonoids did not substantially inhibit pentoxyresorufin O-deethylation (CYP2B 1), mephenytoin 4-hydroxylation (CYP2C19), dextromethorphan O-demethylation (CYP2D6), and chlorzoxazone 6-hydroxylation (CYP2E1) activities (IC50: >50 microM). Baicalein and 2',5,6',7-tetrahydroxyflavone inhibited hepatic testosterone 6beta-hydroxylation (CYP3A4) activity with a IC50 of 17.4 and 7.8 microM, respectively. Oroxylin A inhibited diclofenac 4-hydroxylation (CYP2C9) activity with a IC50 of 6.7 microM. In contrast, all flavonoids tested inhibited hepatic caffeine N'-demethylation (CYP1A2) with IC50 values ranging from 0.7 to 51.3 microM. Kinetic analysis revealed that the mechanism of inhibition varied according to the flavonoids. These results suggest that flavonoids tested are inhibitors of hepatic CYP1A2 and that the extracts of Scutellariae radix, widely used as a hepatoprotective agent, may protect the liver through the prevention of CYPIA2-induced metabolic activation of protoxicants. PMID- 11936219 TI - Immunotoxic effects of 2-bromopropane in male Sprague-Dawley rats: a 28-day exposure study. AB - Immunotoxic effects of 2-bromopropane were investigated in male Sprague-Dawley rats. The rats were treated orally daily with 2-bromopropane at 100, 330, or 1000 mg/kg for 28 consecutive days. Four days before necropsy, the rats were immunized intravenously with sheep red blood cells (SRBCs). The body and thymus weights were significantly reduced by treatment with 2-bromopropane at the highest dose. In addition, the numbers of splenic and thymic cells were decreased by 2 bromopropane. In hematology, the numbers of white blood cells, red blood cells, and platelets were significantly reduced. Among the serum clinical parameters, the levels of chloride ion were significantly increased by 2-bromopropane. The antibody response to SRBCs was significantly suppressed at the highest dose. With immunized animals, immunophenotyping of splenic and thymic cells was performed to investigate the changes of the number of macrophages, B cells, and T cells in spleen and the number of CD4+ and CD8+ cells in thymus. The numbers of most cell types were significantly decreased in the spleen when animals were treated with 2 bromopropane at 1,000 mg/kg. Likewise, all cell types of thymus were significantly decreased by 2-bromopropane. The present results suggest that 2 bromopropane may have an immunotoxic potential in male Sprague-Dawley rats when the rats are exposed for 28 d. PMID- 11936220 TI - Maternal-fetal disposition of bisphenol a in pregnant Sprague-Dawley rats. AB - This study describes the maternal-fetal disposition of bisphenol A and its distribution into the placenta and amniotic fluid after iv injection (2 mg/kg) to pregnant Sprague-Dawley rats. Bisphenol A was distributed extensively to the placenta and fetus, with their respective AUC values 4.4- and 2.2-fold greater than AUC for the maternal serum. In contrast, the distribution of bisphenol A into the amniotic fluid was low, with the mean amniotic fluid-to-maternal serum AUC ratio of 0.2. The decay curves of bisphenol A in the placenta, fetus, and amniotic fluid paralleled that of the maternal serum during the terminal elimination phase. A five-compartment open model consisting of the maternal central, maternal peripheral, placental, fetal, and amniotic fluid compartments was used to describe the disposition of bisphenol A in pregnant rats, with the elimination occurring from the maternal central and fetal compartments. Based on this model, bisphenol A delivered to the placenta was transferred primarily to the fetus [kpf/(kpf + kpc + kpa) = 65.4 %], with the remaining fraction transported to the maternal central (33.2%) and amniotic fluid (1.4%) compartments. Bisphenol A was eliminated from the amniotic fluid by the fetal (63.9%) and placental (36.1%) routes. On the other hand, bisphenol A was eliminated from the fetus primarily by the placental route back to mother [kfp/(kfp + kfa + kfo) = 100%], with the amniotic route playing an insignificant role in fetal elimination. The percent contribution of the fetal elimination to the total elimination in the maternal-fetal unit was 0.0% [CLfoAUCfetus/(CLcoAUCmaternal serum + CLfoAUCfetus)]. The pharmacokinetic model used in this study provides insights into the routes of elimination of bisphenol A in the maternal-fetal rat upon maternal administration. PMID- 11936221 TI - Inhibitory effect of peroxisome proliferator-activated receptor gamma agonist on ochratoxin A-induced cytotoxicity and activation of transcription factors in cultured rat embryonic midbrain cells. AB - The effects of 15-deoxy-delta12,14-prostaglandin J2 (15-deoxy PGJ2) on ochratoxin A (OTA)-induced neurotoxicity and on the activation of transcription factors activator protein-1 (AP-1) and nuclear factor-kappa B (NF-kappaB) were investigated in cultured rat embryonic midbrain cells. Twelve-day rat embryo midbrain cells were cultured for 48 h. OTA (0.5 or 1 microg/ml) and/or 1.5-deoxy PGJ2 (0.5 microM) were then added for 48 h. Cell number and neurite outgrowth were determined to assess the neurotoxicity of OTA. AP-1 and NF-kappaB activation was determined by gel mobility shift assay after 3 h of exposure to OTA and/or 15 deoxy PGI2. OTA caused concentration-dependent reductions in neurite outgrowth and cell number, and induced AP-1 and NF-kappaB activation. Cotreatment with 15 deoxy PGJ2 (0..5 microM) blocked OTA-induced decrease in neurite outgrowth and cell number and inhibited AP-1 and NF-kappaB activation. 15-Deoxy PGJ2 (0.5 microM) caused the expression of peroxisome proliferator-activated receptor-gamma (PPAR-gamma) in the cells. Results show that 1.5-deoxy PGJ2 blocked OTA-induced neurotoxicity by inhibiting AP-1 and NF-kappaB activation in cultured rat embryonic midbrain cells. PMID- 11936222 TI - Comparative evaluation of alkylphenolic compounds on estrogenic activity in vitro and in vivo. AB - This study was undertaken to compare the sensitivity of screening test methods and to investigate the structure-activity relationships of the estrogenic activity of alkylphenolic compounds (APs) using in vitro and in vivo assays. Two in vitro systems, MCF-7 cell proliferation (E-screen assay) and competitive binding assay to estrogen receptor (ER), were selected to evaluate the estrogenic effects. Uterotrophic assay and Calbindin-D9K (CaBP9K) mRNA expression were also examined in ovariectomized Sprague-Dawley female rats. A series of APs with various alkyl groups were examined, namely, 4-propylphenol, 4-butylphenol, 4-t butylphenol, 4-pentylphenol, 4-nonylphenol, 4-octylphenol, 4-t-octylphenol, and 4 phenylphenol, and 17beta-estradiol (E2) was used as a positive control. In the E screen assay, E2 was found to induce maximum proliferation of MCF-7 cells at 1 nM. Among the APs, 4-t-octylphenol and 4-nonylphenol were found to be considerably more potent than any other compound and estrogenic effects were detectable at 1 and 10 microM, respectively. 4-t-Octylphenol and 4-nonylphenol inhibited the binding of E2 to the ER of MCF-7 cells in a competitive ER binding assay. The uterotrophic effects to APs (10, 50, 200, and 400 mg/kg/d) were compared to E2 (1 microg/kg) in ovariectomized rats after treatment for 3 d. 4 Nonylphenol, 4-t-octylphenol, and 4-phenylphenol produced dose-dependent increases in the uterine weights of ovariectomized rats. In the CaBP-9K mRNA expression test, CaBP-9K mRNA levels were detected in the uteri of ovariectomized rats treated with 4-pentylphenol (400 mg/kg), 4-nonylphenol, 4-phenylphenol (200 and 400 mg/kg), and 4-t-octylphenol (50 mg/kg and above), respectively. In the dot blot assay, CaBP-9K mRNA levels were significantly increased in rats exposed to 4-t-octylphenol (200 and 400 mg/kg), 4-pentylphenol, 4-nonylphenol, and 4 phenylphenol (400 mg/kg), respectively. Among the APs, compounds with bulky alkyl groups or higher carbon numbers possessed higher estrogenic capacity. In addition, the pattern of CaBP-9K expression correlated with that of the 3-d uterotrophic assay. Therefore, our results suggest that the CaBP-9K gene might be used as a potential biomarker for the screening of endocrine disruptors. PMID- 11936223 TI - Effects of flutamide on puberty in male rats: an evaluation of the protocol for the assessment of pubertal development and thyroid function. AB - To establish a test protocol for the rodent 20-d thyroid/pubertal assay, flutamide, a non-steroidal androgen antagonist, was administered to intact male Sprague-Dawley rats from postnatal d 33 for 20 d, and several reproductive endpoints were examined to assess the sensitivity of a number of parameters with respect to the detection of endocrine-related effects. Immature male rats were divided into 4 groups and given flutamide once daily by oral gavage at doses of 0, 1, 5, or 25 mg/kg/d. Prepuce separation was significantly delayed in flutamide treated rats (5 and 25 mg/kg/d). One day after the last dose, the rats were sacrificed. Flutamide treatment resulted in a significant reduction in the weights of epididymides, ventral prostate, seminal vesicles plus coagulating glands and fluid, levator ani plus bulbocavernosus muscles, Cowper's glands, and glans penis. The weight of adrenal glands decreased at 25 mg/kg/d, while testes and any other organ weights were unaffected. No microscopic changes were observed in the thyroid glands. Serum levels of testosterone were significantly increased in the flutamide-treated groups (5 and 25 mg/ kg/d) and serum levels of estradiol were also increased (25 mg/kg/d). No differences were observed in the serum thyroxine levels. These results indicate that flutamide delays puberty in the male rat, and its mode of action appears to be via altered secretion of steroids, which subsequently affect the development of the reproductive tract. Thus, this assay might be used as an alternative for screening antiandrogenic activities of chemicals. PMID- 11936224 TI - DI-(2-ethylhexyl) phthalate-induced cell proliferation is involved in the inhibition of gap junctional intercellular communication and blockage of apoptosis in mouse Sertoli cells. AB - Di-(2-ethylhexyl) phthalate (DEHP) has been studied on gap junctional intercellular communication (GJIC) and apoptosis in cultured normal mouse Sertoli cells. Since the inhibition of GJIC and programmed cell death or apoptosis play important roles in tumor promotion and developmental toxicity, it has been hypothesized that tumor promoters may inhibit apoptosis by blocking GJIC. The results showed that the most significant downregulation of GJIC was detected at 9 h after DEHP treatment. However, a significant concentration-dependent pattern was not observed at concentrations of 100 and 500 microM, but there was a time dependent recovery of GJIC. DEHP inhibited the apoptotic changes in the cells such as chromatin condensation, nuclear fragmentation, and the cleavage of poly(ADP-ribose) polymerase. Morphological changes related to apoptosis appeared in the nontreated cells after 12 h of serum deprivation. These morphological changes were significantly reduced in the TM5 Sertoli cells treated with 500 microM DEHP for 24 h. These results suggest that DEHP inhibited apoptosis in this cell line, preceded by the downregulation of GJIC. It was also found that DEHP reduced the phosphorylation of Cx43, which might partly explain the mechanism of inhibition of GJIC. PMID- 11936225 TI - Effects of dibutyl phthalate and monobutyl phthalate on cytotoxicity and differentiation in cultured rat embryonic limb bud cells; protection by antioxidants. AB - This present study was undertaken to examine the effects of DBP and its metabolite mono-n-butyl phthalate (MBuP) on cytotoxicity and differentiation in cultured rat embryonic limb bud cells. When limb bud cells extracted from rats on gestation d 12.5 were treated with DBP or MBuP for 96 h, induction of cytotoxicity and inhibition of cell differentiation were observed in a concentration-dependent manner. However, MBuP elicited a toxic effect at higher concentrations than DBP. The IC50 values of DBP for cytotoxicity (measured by neutral red uptake) and cell differentiation (measured by alcian blue staining) were 25.54 microg/ml (91.75 microM) and 21.21 microg/ml (76.20 microM), respectively. The IC50 values of MBuP for cytotoxicity and cell differentiation were 307.24 microg/ml (1.38 mM) and 142.61 microg/ml (0.64 mM), respectively. in order to determine whether free radicals are related to induction of cytotoxicity and inhibition of differentiation by DBP in limb bud cells, DBP was coadministered with several antioxidants, including catalase and vitamin E acetate to limb bud cells. Cotreatment with catalase and vitamin E acetate decreased induction of cytotoxicity and inhibition of differentiation by DBP in limb bud cells. However, these compounds did not show any protective effect against MBuP. Results indicate that DBP and MBuP induced developmental toxicity in rat embryonic limb bud cells and suggest that this effect of DBP might be exerted through oxidative stress. PMID- 11936226 TI - Iron enhancement of oxidative DNA damage and neuronal cell death induced by salsolinol. AB - A group of naturally occurring isoquinoline alkaloids have been detected in certain regions of mammalian brain. One such compound is salsolinol (SAL; 1 methyl-6, 7-dihydroxy-1,2,3,4-tetrahydroisoquinoline). This endogenous isoquinoline derivative has been considered to be implicated in the pathophysiology of chronic alcoholism and Parkinsonism. The present study deals with the DNA strand scission induced by SAL in the presence of iron. Incubation of phiX174 DNA with SAL and ferric ion led to conversion of the supercoiled DNA to open circular and linear forms, which was inhibited by the iron chelator deferoxamine, catalase, and scavengers of reactive oxygen species. SAL in combination with Fe(III) also produced 8-hydroxydeoxyguanosine in calf thymus DNA. Exposure of PC12 cells to SAL produced concentration-dependent reduction in viability, which was exacerbated by iron and ameliorated by deferoxamine. PMID- 11936227 TI - Alice M. Horowitz, Ph.D., oral health change agent: an unsung hero. PMID- 11936228 TI - Major long-term factors influencing dental education in the twenty-first century. AB - With evolutionary advances in oral science over the long term, clinical reliance on chemotherapeutics, bacterial replacement therapy, and immunization will necessitate a broader background in medicine. The dramatic increase in the old age population will also require a much stronger medical background. By 2050, those over fifty-five years of age will represent 56 percent of the population, and 25 percent of these will be sixty-five years of age and older. The merging of dental and medical education is predicted to occur within the twenty-first century. Other topics addressed include research activities, with recommended strategies to enhance the integration of scientific and clinical dental approaches; the problem of dental faculty isolation; and the implications of financial constraint and accountability. PMID- 11936229 TI - Correlation of admission criteria with dental school performance and attrition. AB - This study was conducted to provide current information on the relationship between admission criteria and dental school performance, including the association of admission criteria and dental school outcomes such as remediation and attrition. Standard tests of bivariate association and multivariate regression models appropriate for continuous and discrete dependent variables were used to examine the relationship between multiple indicators of admission criteria and dental school performance for six recent classes at the University of Florida College of Dentistry (UFCD). The admission criteria included the undergraduate science grade point average (GPA), undergraduate non-science GPA, Dental Admissions Test (DAT) academic score, Perceptual Motor Aptitude Test (PMAT) score, and admission interview score. Measures of dental school performance were the National Dental Board Examination Part I and Part II (NB-I, NB-II) scores, yearly and final dental school GPA, and academic progress through the UFCD program. In general, most admission criteria were good bivariate indicators of dental school performance. Multivariate analyses indicated that students with higher undergraduate science GPAs and DAT academic scores were more likely to achieve higher NB-I and NB-II scores. The undergraduate science GPA and admission interview score were the most consistent determinants of dental school GPA. Students with lower undergraduate science GPAs, DAT academic scores, and PMAT scores were more likely to remediate, to repeat an academic year, or to be dismissed. Although bivariate differences were observed in several admission criteria of students who remediated one or more courses, repeated an academic year, or were dismissed only the undergraduate science GPA and the PMAT score were indicators of programmatic progress in the multivariate analysis. PMID- 11936231 TI - Phase 1 of a comprehensive course review process: program innovation. AB - A systematic college-wide course review process can serve as an effective and efficient vehicle for both curricular management and faculty development. The purpose of this paper is to describe Phase I of an evolving course review process at the University of Kentucky College of Dentistry (UKCD). Phase 1 was designed to provide the foundation necessary to ultimately achieve an overall program of continuous evaluation that will produce sustainable curricular improvement and faculty renewal. The course review process includes elements of self-assessment and collaborative peer review. Elements of the current system include emphasis on both the curriculum as a whole and individual course characteristics. It encourages faculty to view their courses as part of the whole and to develop sensitivity to the impact their courses have on other parts of the curriculum. Equally important are the elements that focus on individual course elements and educational methodologies used. PMID- 11936230 TI - Evaluation of web-based dental CE courses. AB - Various organizations offer online continuing dental education (CDE) courses. While previous investigations focused on objective measures to determine the quality of the courses, this exploratory study evaluates the participants' experience with them. We surveyed 436 past course participants from nine online CDE courses (courses provided by six organizations) regarding their experience with the courses taken. Our analysis of the 169 responses (38.8 percent response rate) focuses on how the participants of online CDE courses can be characterized; whether the participants' expectations were met by the courses; how the participants evaluated the content of the courses; why they enrolled; and the participants' experience of the online environment. The results suggest that online CDE courses partially meet the needs and expectations of dental professionals. The lack of communication with peers and instructors as well as the fact that courses appeared outdated were main reasons for dissatisfaction. Most of the participants accomplished their goals of gaining new knowledge and deepening their understanding of the subject. Based on this evaluation, future courses can be tailored to meet more closely the expectations and needs of dental professionals. PMID- 11936232 TI - Making a comprehensive diagnosis in a comprehensive care curriculum. AB - Comprehensive care models in dental education encourage students to deliver patient-centered care. But to deliver effective comprehensive care, a clinician must first make a comprehensive diagnosis. Students of general dentistry are taught to make one or more diagnoses as defined by the dental specialties, and to direct patient care accordingly. Without a comprehensive diagnosis, patients may receive fragmented, poorly prioritized care that is inappropriate to their overall oral health. This paper presents a simple diagnostic classification that can be used to make a comprehensive diagnosis with which to guide the student of general dentistry in planning comprehensive care. PMID- 11936233 TI - Survey of undergraduate esthetic courses in U.S. and Canadian dental schools. AB - U.S. and Canadian dental schools were surveyed regarding curriculum issues related to undergraduate dental esthetic restorative courses. A one-page survey instrument was sent to deans of academic affairs (n=59) of dental schools to complete or forward to the most appropriate faculty at their respective schools who would be knowledgeable about the esthetic restorative curriculum. Responses were received from forty-two dental schools (42/59) for a response rate of 72.9 percent. The first part of the survey asked if a specific esthetic restorative course was offered at their school, if it was mandatory or elective, and details of the course length and content. The second part of the survey asked whether selected esthetic restorative procedures were included in the curriculum. Respondents to this survey indicated that most types of esthetic restorative procedures are taught whether or not an esthetic course is included in the curriculum. PMID- 11936234 TI - Tobacco control and prevention effort in dental education. PMID- 11936235 TI - Applicant analysis: 2000 entering class. AB - There were 7,770 applicants to the entering dental school class of 2000. This is almost 14 percent less than the number of applicants to the entering class of 1999. Since the peak of dental school applicants in 1997 (at 9,829), the number has declined 21 percent. (This is most similar to the decline that has occurred in medical school applicants since their peak of applicants in 1996, at 46,968.) Almost 55 percent of the applicants to dental school were enrolled in 2000. Dental schools reported 4,234 first-time, first-year enrollees in 2000. This is an increase of 25 enrollees over the number reported in 1999. Since 1989, when dental school enrollment once again began to increase, total first-year dental school enrollment has increased 8.7 percent. The number of applicants per first time, first-year position was 1.84 in 2000. It was 2.14 in 1999. (The most recent low was 1.34 in 1989.) The GPA and DAT scores of the first-time, first-year enrollees in 2000 were all either equal to or slightly higher than they were in 1999. Women were approximately 40 percent of the applicants and first-time, first year enrollees in 2000, up slightly from 1999. Underrepresented minorities comprised slightly over 12 percent of the applicants and 10.6 percent of the first-time, first-year enrollees, also up slightly from 1999. PMID- 11936236 TI - Use of mental health services by the rural aged: longitudinal study. AB - Recent longitudinal research investigating historic shifts in attitudes toward mental health services reveal a positive cohort shift. Most likely, this shift equates to greater use of mental health services. The purpose of this study was to examine historic shifts in the use of mental health services in a rural population. In 1987, we administered a survey to 358 randomly selected rural individuals investigating their willingness to use mental health services. In 1994, we resurveyed 110 individuals from our original sample. As predicted, we found that there was a shift in our participants' willingness to use mental health services, increasing from 5% to 18%. This increased use, although improved, still fell somewhat below the actual need for mental health services, which had remained constant (about 25%). Interestingly, there was no greater reliance on psychotropic medication in lieu of psychotherapy (about 10%). In 1987, we found that those rural elderly who had the greatest need for mental health services were the least likely to use them. This trend may be shifting as well, although it requires further investigation and confirmation. The salient implications for the design and implementation of rural mental health services for baby boomers are noted. PMID- 11936237 TI - The "time and change" test: an appropriate method to detect cognitive decline in the elderly. AB - The Time and Change (T&C) test is an easy and time-saving test validated for the detection of dementia. Our aim was to determine how geriatric features like depression, disability, and comorbidity are able to influence the result of the T&C and, consequently, to decide whether it could be a reliable screening test for cognitive impairment in the elderly. A total of 220 participants (mean age = 75.8+/-9.6 years, 63.7% females) underwent the T&C, Mini-Mental State Examination, and the Clock Drawing Test; Activities of Daily Living, Instrumental Activities of Daily Living, comorbidity, and depression were also evaluated. Time and Change-positive participants were older, had poorer cognitive tests, and had higher levels of disability and comorbidity than participants testing negative. Multivariate analysis showed that cognitive impairment and comorbidity were the only features that influenced the T&C, regardless of age, education, disability, and depression. We conclude that the T&C should be implemented in primary care because it quickly identifies elderly patients with cognitive impairment who need a more accurate evaluation. PMID- 11936238 TI - Which level of care is preferred for end-stage dementia? Survey of Taiwanese caregivers. AB - In Western countries, most family caregivers view some degree of palliative care as appropriate for severely demented care recipients. In Asian countries, caregivers' attitudes toward such an important issue have not been sufficiently investigated. Therefore, we surveyed Taiwanese caregivers' attitudes toward the terminal care of demented care recipients by questionnaire. In 115 (48 men, 67 women; mean age 73.7+/-9.5 years) caregivers who completed the questionnaire, most caregivers (61%) preferred highly aggressive care, whereas only 3% preferred solely palliative care at the end of the demented care recipients' life. Caregivers having a lower education level (< or = 12 vs > 12 years) or having care recipients of a younger age (< or =75 vs >75 years) exhibited a higher preference for cardiopulmonary resuscitation (CPR) (P = .020 and P = .010, respectively). Sixteen percent of caregivers accepted postmortem autopsy for their care recipients. Twenty-eight percent of caregivers of home-resident recipients anticipated institutionalization, especially those with care recipients having moderate to severe stages of dementia. Co-surrogate caregivers, including the spouse and the offspring, were the most common proxies (39%) for care recipients with end-stage dementia. By comparison with Western countries, the low acceptance rate of CPR refusal and nursing home placement by our caregivers might result from cultural differences. PMID- 11936239 TI - Lack of association between the apolipoprotein E genotype and depression in Alzheimer's disease. AB - The epsilon4 allele of apolipoprotein (apo E) is one of the risk factors for late onset Alzheimer's disease (AD). We evaluated the association between apo E genotypes and depression in patients with AD. A psychiatrist interviewed all patients and their caregivers for depression using a Chinese version of the Structured Clinical Interview for the Diagnostic and Statistical Manual of Mental Disorders, Third Edition, Revised, and for the severity of depression using the Hamilton Depression Rating Scale (HDRS). Twenty-five of the 149 patients with AD were diagnosed with depressive disorders. The numbers of patients in each apo E genotype were 10 in epsilon2/3, 2 in epsilon2/4, 74 in epsilon3/3, 46 in epsilon3/4, and 17 in epsilon4/4. We did not find an association between depression and the presence or absence of the epsilon4 or epsilon2 allele. The HDRS scores were not different in patients with AD with the epsilon4 or epsilon2 allele or in those patients without them. Our study did not find an association between depression and the apo E epsilon4 or epsilon2 allele in AD. PMID- 11936241 TI - Relationship between alcohol consumption and Folstein mini-mental status examination scores among older cognitively impaired adults. AB - The purpose of this study was to determine whether older cognitively impaired adults with significant current or past alcohol histories manifest distinctive cognitive profiles as determined by a widely used cognitive screen, the Folstein Mini-Mental Status Examination (MMSE) test, when compared with older persons without significant alcohol histories. Study participants included 801 consecutive patients, ages 65 years or above, who underwent comprehensive geriatric assessments. Proxy-reported current alcohol intake was classified as none, former, light (<1 drink/week), moderate (> or = 1 but < 14 drinks/week), and heavy (> or = 14 drinks/week), and the presence of an alcohol abuse/dependence disorder was determined by medical record review. Potential exposure-outcome associations were assessed for patients (n = 470) with established cognitive impairment, defined as an MMSE score < 24, and for individuals with MMSE scores > or = 24 (n = 331). Among participants with established cognitive impairment, mean (total) MMSE scores were not significantly different (17.2 vs 16.4 vs 18.5 vs 18.5 vs 17.4) across the categories of current alcohol exposure (none, former, light, moderate, and heavy). Mini-Mental State Examination subscores also did not vary as a function of current alcohol consumption, and a history of alcohol abuse/dependence was not associated with differences in total scores or individual MMSE subscores in this patient group. Among participants with MMSE scores > 24, increased current use of alcohol or a history of alcohol abuse/dependence was not associated with lower total MMSE scores or individual MMSE subscores. Despite these negative findings, prospective investigations of older populations that incorporate more comprehensive cognitive measures are warranted because (1) the Folstein is a brief cognitive screen that does not assess all cognitive domains, and (2) alcohol is a demonstrated risk factor for adverse cognitive outcomes in middle-aged adults and is potentially modifiable. PMID- 11936240 TI - Apolipoprotein E and alpha-1-antichymotrypsin genotypes do not predict time to psychosis in Alzheimer's disease. AB - Psychotic symptoms occurring in Alzheimer's disease (AD + psychosis, AD + P) are a marker for a more rapidly deteriorating phenotype. We have developed a polygenic model of AD + P risk, conditioned on the presence of AD. Whether risk genes for AD itself contribute to AD + P risk is not established, although our model predicts they will not. The most important identified genetic determinant of sporadic, late-onset AD is the apolipoprotein E epsilon 4 allele (APOE4). The effect of APOE4 on AD phenotype is to reduce the age of onset of AD. Prior studies examining the association of APOE4 with AD + P have reported conflicting results. However, no prior studies have examined if APOE4 reduces time to onset of psychosis in AD. The objective of this study was to examine the effect of APOE4 and alpha1-antichymotrypsin/AA (ACT/AA) genotypes on time to psychosis onset in subjects with AD. A longitudinal study of psychosis incidence in 316 subjects with AD with no history of current or prior psychotic symptoms at entry was undertaken. APOE and ACT genotyping was conducted per established protocols. Data were analyzed by survival analysis and Cox proportional hazards models. There were no significant associations of APOE or ACT genotypes with time to psychosis onset and no significant interaction of these genotypes with time to psychosis onset. There remained no significant associations after covarying for age, age of AD onset, degree of cognitive impairment, gender, race, and education. This is the first study to examine the genetic prediction of psychosis onset in AD. The findings support the hypothesis that these two genetic determinants of AD risk do not contribute to the risk of development of psychotic symptoms in AD. PMID- 11936242 TI - "Life is not worth living": hopelessness in Alzheimer's disease. AB - This study investigated the prevalence and clinical correlates of hopelessness among 91 patients diagnosed with probable Alzheimer's disease (AD) according to National Institute of Neurological Communicative Disorders and Stroke-Alzheimer's Disease and Related Disorders Association criteria. Hopeless ideation was measured with Item 3 on the Hamilton Depression Rating Scale (Suicide), which inquires specifically whether the patient thinks "life is not worth living." The results showed that hopelessness was present in 10% (n = 9) of the sample. Patients with these cognitions evidenced greater psychological symptoms of mood disturbance and more insight into their cognitive and functional impairments. Unrelated factors included age, education, Mini-Mental State Examination score, neurovegetative signs of depression, affective expressions of depression, agitation/disinhibition, and psychosis. Although indifference is frequent in AD, these results indicate that thoughts of hopelessness are also a common phenomenon, occurring in approximately 10% of our probable AD cohort. These thoughts appear to be related to the subjective expressions of depression and anxiety rather than the neurovegetative or affective signs and are more prominent among patients with greater deficit awareness. PMID- 11936243 TI - No melancholia in poststroke depression? A phenomenologic comparison of primary and poststroke depression. AB - The present study aimed at the phenomenologic comparison of depressive symptoms in elderly patients with poststroke depression (PSD) or primary depression (depression without a known neuropathology). We investigated 20 patients with PSD and 41 patients with primary depression. A structured clinical interview based on the Diagnostic and Statistical Manual of Mental Disorders, Fourth Edition, self- and observer-based depression rating scales, a clinical neurologic examination, and neuroradiologic analysis based on standardized computed tomographic scan analysis were applied. The Cornell Depression Scale was used to compare mood related signs, behavioral disturbances, physical signs, disturbances of cyclic signs, and ideational disturbances in both groups of patients. Those with PSD exhibited no melancholia and fewer cyclic and ideational disturbances but more physical signs of depression. These findings are in line with those of endocrinologic studies. Differences between both groups of patients indicate that careful considerations and further research are needed before treatment strategies developed for and evaluated in patients with primary depression are applied to patients with PSD. PMID- 11936244 TI - The global deterioration scale: relationships to neuropsychological performance and activities of daily living in patients with vascular dementia. AB - In the present study, we examined the relationships between ratings on the Global Deterioration Scale (GDS) and activities of daily living and cognitive function in 39 individuals with vascular dementia (VaD). The results of the study revealed significant correlations between GDS rating and performance on cognitive tests, including memory and overall cognitive ability. In addition, the GDS was significantly related to ratings of instrumental activities of daily living. Comparisons between patients with VaD with GDS scores between 4 and 6 (n = 21) and patients with scores between 2 and 3 (n = 18) revealed greater cognitive and functional deficits in the group with higher GDS scores. Further, the GDS score accurately classified 87% of the patients with VaD. These findings provide support for the validity of the GDS in general staging of dementia severity of VaD. PMID- 11936245 TI - Decline in long-term circadian rest-activity cycle organization in a patient with dementia. AB - Continuous measurement of the circadian rest-activity cycle for 598 days in a demented patient with probable Alzheimer's disease revealed slow progressive changes in temporal organization until death. Circadian and sleep analysis of the actigraphic data provided objective documentation of the gradual insertion of wakefulness into rest and rest into wake periods. Pacing, a nonphotic zeitgeber strengthening, led to improved synchronization of the rest-activity cycle. PMID- 11936246 TI - Blue rubber bleb nevus syndrome in a patient with ataxia and dementia. AB - Blue rubber bleb nevus syndrome (BRBNS), an uncommon disorder characterized by cavernous hemangiomas, most often of the skin and gastrointestinal tract, is usually diagnosed during childhood and young adulthood. We made this diagnosis in an octogenarian referred to a geriatric medicine clinic because of concerns about his ability to live independently. Ataxia, dementia, focal neurologic signs, and bluish/purplish vascular nodules on his lips, buccal mucosa, tongue, chest, and neck were noted on physical examination. Magnetic resonance imaging (MRI) revealed an old left parietal infarction, multiple cavernous hemangiomas most densely concentrated in the subcortical structures and cerebellum, and areas of hemosiderin deposition. Skin biopsy findings were consistent with hemangioma. The physical examination, MRI, and skin biopsy made a diagnosis of BRBNS likely. The patient's ataxia, dementia, and other neurologic signs can be explained by previous hemorrhage from the vascular malformations in his brain. Blue rubber bleb nevus syndrome is an uncommon cause of a relatively common geriatric syndrome presentation. PMID- 11936247 TI - Within-subject comparison of real and hypothetical money rewards in delay discounting. AB - A within-subject design, using human participants, compared delay discounting functions for real and hypothetical money rewards. Both real and hypothetical rewards were studied across a range that included $10 to $250. For 5 of the 6 participants, no systematic difference in discount rate was observed in response to real and hypothetical choices, suggesting that hypothetical rewards may often serve as a valid proxy for real rewards in delay discounting research. By measuring discounting at an unprecedented range of real rewards, this study has also systematically replicated the robust finding in human delay discounting research that discount rates decrease with increasing magnitude of reward. A hyperbolic decay model described the data better than an exponential model. PMID- 11936248 TI - Evidence against a constant-difference effect in concurrent-chains schedules. AB - Savastano and Fantino (1996) reported that in concurrent-chains schedules, initial-link choice proportions remained constant as terminal-link durations increased as long as the subtractive difference between the two terminal-link schedules remained constant. Two experiments with pigeons were conducted to examine this constant-difference effect. Both experiments used equal variable interval schedules as initial links. The terminal links were fixed delays to reinforcement in Experiment 1 and variable delays to reinforcement in Experiment 2. The durations of the terminal links were varied across conditions, but the difference between pairs of terminal links was always 10 s. In both experiments, preference for the shorter terminal link became less extreme as terminal-link durations increased, so a constant-difference effect was not found. It is argued, however, that this choice situation does not provide clear evidence for or against delay-reduction theory versus other theories of choice. PMID- 11936250 TI - The generality of selective observing. AB - Four rats obtained food pellets by poking a key and 5-s presentations of the discriminative stimuli by pressing a lever. Every 1 or 2 min, the prevailing schedule of reinforcement for key poking alternated between rich (either variable interval [VI] 30 s or VI 60 s) and lean (either VI 240 s, VI 480 s, or extinction) components. While the key was dark (mixed-schedule stimulus), no exteroceptive stimulus indicated the prevailing schedule. A lever press (i.e., an observing response), however, illuminated the key for 5 s with either a steady light (S+), signaling the rich reinforcement schedule, or a blinking light (S-), signaling the lean reinforcement schedule. One goal was to determine whether rats would engage in selective observing (i.e., a pattern of responding that maintains contact with S+ and decreases contact with S-). Such a pattern was found, in that a 5-s presentation of S+ was followed relatively quickly by another observing response (which likely produced another 5-s period of S+), whereas exposure to S- resulted in extended breaks from observing. Additional conditions demonstrated that the rate of observing remained high when lever presses were effective only when the rich reinforcement schedule was in effect (S+ only condition), but decreased to a low level when lever presses were effective only during the lean reinforcement component (S- only condition) or when lever presses had no effect (in removing the mixed stimulus or presenting the multiple-schedule stimuli). These findings are consistent with relativistic conceptualizations of conditioned reinforcement and extend the generality of selective observing to procedures in which the experimenter controls the duration of stimulus presentations, the schedule components both offer intermittent food reinforcement, and rats serve as subjects. PMID- 11936249 TI - Disruption of temporally organized behavior by morphine. AB - Four pigeons pecked keys in two different procedures commonly used in the study of timing, or temporal discrimination. Sessions consisted of 40 trials. During half of the trials, two keys were presented for 50 s. Left-key pecks were reinforced according to a variable-interval 67.86-s schedule during the first 25 s of the trial, and right-key pecks were not reinforced. During the second 25 s of the trial, right-key pecks were reinforced according to the same schedule, and left-key pecks were not reinforced. In the other half of the 40-trial session, the center key was presented. The majority of these trials arranged fixed interval 2.5-s schedules. Occasionally a probe, or peak-interval, trial was presented. These trials were 100 s in duration and terminated without reinforcement. These two procedures were used to examine the effects of morphine on indexes of timing and on patterns of responding. Morphine altered behavior in a race-dependent manner in both procedures. Low baseline (saline) response rates were increased following morphine administration, and high baseline rates were either unaffected or decreased slightly. Rate-dependent effects appeared as leftward shifts in the timing index for two-key trials and decreases in the index of curvature for fixed-interval trials. Despite large changes in response rates, no consistent shift of the peak time was observed during peak-interval trials. These results are discussed primarily in terms of rate dependency; that is, rates of responding following drug administration tend to be determined in large part by rates of responding under baseline conditions. PMID- 11936251 TI - High-probability stimulus control topographies with delayed S+ onset in a simultaneous discrimination procedure. AB - Experimenters and teachers use discrimination learning procedures to encourage reliable attending to stimulus differences defined as relevant for their purposes. Put another way, the goal of discrimination training is to establish high-probability stimulus control topographies that are coherent with experimenter or teacher specifications. The present research was conducted to investigate a novel procedure for encouraging stimulus control topography coherence. Participants were 13 adolescents with severe intellectual handicaps. During an initial Condition A, all were exposed to a simultaneous discrimination procedure. Participants could select a form alternating with a black field (S+) or an identical form that did not alternate (S-). Accuracy scores were typically low, and there was little evidence of coherent stimulus control topographies. Subsequently, the procedure was changed. During Condition B, every trial initially presented two identical nonalternating S- forms (Trial State 1). If the participant made no selection for 5 s, one of the forms began to alternate with the black field, and he or she could make the S+/S- discrimination (Trial State 2). Selections during Trial State I prolonged the delay to Trial State 2 until there had been no response for 5 s. During Condition B, S+/S- discrimination accuracy scores improved rapidly and markedly for most participants. Reinstating Condition A often resulted in diminished accuracy scores. This study thus (a) demonstrated a novel procedure for encouraging stimulus control topography coherence and (b) provided support for the interpretation that intermediate accuracy scores may be due to different topographies of stimulus control that co occur in the same discriminative baseline. PMID- 11936252 TI - Time of supplemental feeding alters the effects of cocaine on lever pressing of rats. AB - The present experiment assessed the effects of cocaine on the lever pressing of 4 rats maintained during 15-min sessions by a fixed-ratio 50 schedule of food reinforcement. Across phases, supplemental food was provided either immediately or 2 hr after sessions. Two rats began the experiment in the delayed-feeding condition, and 2 began the experiment in the immediate-feeding condition. Rates of lever pressing of 2 rats sometimes decreased to low levels near the ends of sessions when supplemental feeding was provided immediately, but were consistently high throughout sessions when supplemental feeding was delayed. Cocaine (1.0 to 17.0 or 30.0 mg/kg) was administered intraperitoneally 15 min prior to test sessions. In most cases, cocaine suppressed response rates at lower doses under immediate-feeding conditions. Decreases in overall response rates were correlated with dosedependent increases in the time rats spent not responding. It is suggested that delaying the time of postsession feeding increased response strength, as indicated by greater resistance to the rate suppressive effects of cocaine. PMID- 11936253 TI - Magnetite-alginate beads for purification of some starch degrading enzymes. AB - Starch degrading enzymes, viz., beta-amylase, glucoamylase, and pullulanase, were purified using magnetite-alginate beads. In each case, the enzyme activity was eluted by using 1.0 M maltose. beta-Amylase (sweet potato), glucoamylase (Aspergillus niger), and pullulanase (Bacillus acidopullulyticus) from their crude preparations were purified 37-, 31-, and 49-fold with 86, 87, and 95% activity recovery, respectively. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis analysis showed single band in each case. PMID- 11936254 TI - CaMV 35S promoter directs beta-glucuronidase expression in Ganoderma lucidum and Pleurotus citrinopileatus. AB - The cauliflower mosaic virus (CaMV) 35S promoter has been most commonly used in plant transformation studies, but its activity in mushrooms has not been reported. p301-b is a binary vector containing a bialaphos resistance gene driven by the promoter of Lentinus edodes glyceraldehyde-3-phosphate dehydrogenase (GPD) gene. CaMV 35S-GUS was inserted into p301-b, and the resulting construct p301-bG was transformed to protoplasts of Ganoderma lucidum and basidiospores of Pleurotus citrinopileatus. GUS activity was observed in the transformants, indicating that CaMV 35S promoter can direct expression of exogenous gene in the mushrooms. This is the first report on the application of CaMV 35S promoter in genetic modification of mushrooms. PMID- 11936255 TI - Human marrow-derived mesenchymal progenitor cells: isolation, culture expansion, and analysis of differentiation. AB - A number of adult mesenchymal tissues contain subpopulations of undifferentiated cells, which retain the capacity to differentiate along multiple lineages. These mesenchymal progenitor cells may be cultured in an undifferentiated state and, when given the appropriate signals, differentiate into an expanding list of several mesenchymal and a few ectodermal derived tissues. The maintenance and propagation of the multipotential nature of these progenitor cell populations are crucially dependent on the isolation protocol, the culture expansion conditions, particularly the properties of the fetal bovine serum supplement in the culture medium. This article describes a method for selection of the appropriate serum lot, and introduces a simplified isolation technique to optimize the yield of progenitor cells that maintain the capability of undergoing multilineage differentiation in response to appropriate cues. Cell populations isolated and culture expanded in this manner, by virtue of their multiple differentiation potential, should serve as ideal candidate cells for tissue engineering applications for the repair and regeneration of tissue damaged by disease and or trauma. PMID- 11936256 TI - A modified nick translation method used with FISH that produces reliable results with archival tissue sections. AB - Nick translation is used to label DNA and RNA to produce probes for in situ hybridization and Northern and Southern blotting. Fluorescence in situ hybridization (FISH) is a widely applied technique used to determine chromosomal and genetic anomalies in many biological samples. Initially the technique was applied to metaphase preparations, but the usefulness of detecting genetic anomalies in solid tumors in situ has resulted in the development of modified protocols. Formalin fixed paraffin processed tissue sections present novel challenges when applying FISH; the probes must be small (between 200 and 600 base pairs) and pretreatment is necessary before the probes can be applied to tissue sections, to promote probe access to target DNA. Here we report on a modification of a nick translation method to produce a probe that can reliably be used with FISH in paraffin processed tissue sections. PMID- 11936257 TI - Resistance to tetracycline, macrolide-lincosamide-streptogramin, trimethoprim, and sulfonamide drug classes. AB - The discovery and use of antimicrobial agents in the last 50 yr has been one of medicine's greatest achievements. These agents have reduced morbidity and mortality of humans and animals and have directly contributed to human's increased life span. However, bacteria are becoming increasingly resistant to these agents by mutations, which alter existing bacterial proteins, and/or acquisition of new genes, which provide new proteins. The latter are often associated with mobile elements that can be exchanged quickly across bacterial populations and may carry multiple antibiotic genes for resistance. In some case, virulence factors are also found on these same mobile elements. There is mounting evidence that antimicrobial use in agriculture, both plant and animal, and for environmental purposes does influence the antimicrobial resistant development in bacteria important in humans and in reverse. In this article, we will examine the genes which confer resistance to tetracycline, macrolide-lincosamide streptogramin (MLS), trimethoprim, and sulfonamide. PMID- 11936258 TI - Improving fluorescence-based assays for the in vitro analysis of cell adhesion and migration. AB - Cell adhesion and cell migration are two primary cellular phenomena to be approached in vitro in order to allow for the effective dissection of the individual events and the unravelling of their underlying molecular mechanisms. The use of assays dedicated to the analysis of cell adhesion and migration in vitro also affords an efficient way of conducting larger basic and applied research screenings of the conditions affecting these processes and are potentially exploitable in the context of routine tests in the biological and medical fields. Therefore, there is a substantial interest in devicing more rationale such assays and major contributions in this direction have been provided by the advent of procedures based on fluorescent cell tagging. In this article we describe three fluorescence-based model assays for the qualitative and quantitative assessment of cell adhesion and cell locomotion in static and dynamic conditions. The assays are easily performed, accurate and reproducible, and can be automatized for high-throughput screenings of cell behavior in vitro. Performance of the assays involves the use of certain dedicated disposable accessories, which are commercially available, and a few instruments that, due to their versatility, can be regarded as constituents of a more generic laboratory setup. PMID- 11936260 TI - Universal primers for the PCR-mediated amplification of DNA beta: a molecule associated with some monopartite begomoviruses. AB - DNA beta is an approx 1350 nucleotide, single-stranded DNA molecule which has been shown to be associated with some monopartite geminiviruses of the genus Begomovirus. This component requires the helper begomovirus for replication in the cells of host plants and for insect transmission, possibly by trans encapsidation. Sequence comparisons of the two available DNA beta sequences has identified a highly conserved region upstream of a predicted hairpin structure. Abutting primers designed to this conserved region allows PCR-mediated amplification of the full-length DNA beta component from total nucleic acid extracts isolated from infected plants originating from a variety of geographically distinct sources and host plants. PMID- 11936259 TI - Apoptosis: molecular regulation of cell death and hematologic malignancies. AB - We describe the molecular mechanisms of apoptosis and its relationships with hematologic malignancies, stressing the concept that, both positive and negative deregulation of apoptosis, may be involved in hematologic human diseases. So, this fundamental process must be balanced by so far unknown mechanisms, involving caspases (cysteine proteases, cleaving the protein substrate after an aspartate residue). These, so far known, ten proteases, are interconnected in a molecular cascade, initiated by the release of cytochrome C from mitochondrial membranes and its interaction with APAF-1 (the homolog of the Caenorhabditis e. CED-4) and with caspase 9, that initiates the proteolitic cascade (1,2). The conclusion is that apoptosis is a very important process, but yet poorly known in molecular details, in spite of the efforts of many scientists. Even the role of bcl-2, the main gene protecting from apoptosis, is still unknown. We close this chapter with a list of ten different technical approaches that can be useful tools to study apoptosis, and tracing the molecular principles on which they are based. PMID- 11936261 TI - Dysplasia arising in barrett's esophagus: diagnostic pitfalls and natural history. AB - Barrett's esophagus is a complication of chronic gastroesophageal reflux disease and is defined as a change in the esophageal epithelium of any length that can be recognized at endoscopy and is confirmed to have intestinal metaplasia by biopsy. Esophageal ulcerations and stricture are rarely seen, and the major complications of this disease are epithelial dysplasia and esophageal adenocarcinoma. Dysplasia is felt to represent the best currently available marker of increased cancer risk in these patients. However, there are many pitfalls in the histologic recognition of dysplasia, a particularly difficult problem in the face of active inflammation in patients with ongoing reflux disease. The recognition and grading of dysplasia is subject to significant interobserver variability, particularly at the lower end of the histologic spectrum (negative v indefinite for dysplasia v low-grade dysplasia). High-grade dysplasia and to a lesser degree low-grade dysplasia are markers of increased cancer risk, although their natural history are difficult to determine. Up to 40% of patients with a preoperative diagnosis of high-grade dysplasia have an adenocarcinoma in their esophagectomy specimen. Despite this observation, there is still debate as to whether esophagectomy or close endoscopic surveillance with biopsy is the most appropriate and cost-effective way to manage these patients. The search for more objective surrogate biomarkers that recognize patients who are truly at risk of progressing along the dysplasia carcinoma sequence is underway. However, no biomarker has yet proven to be superior to the histologic recognition of dysplasia in identifying these high risk patients. PMID- 11936262 TI - Esophageal squamous dysplasia. AB - Esophageal squamous dysplasia (ESD) appears to be the most important precursor for squamous cell carcinoma (SCC), and this observation is supported by early molecular findings. Pathologists in high incidence areas of the world, such as China, frequently may encounter ESD during surveillance cytology and biopsy screenings of high risk populations. ESD in low risk areas, such as the United States, is more commonly seen in esophagi resected for squamous cell carcinoma. As at other sites, ESD can be graded histologically depending on the thickness of epithelial involvement and cytologically based largely on nuclear size and chromatin features. Most ESD lesions are endoscopically visible, especially if the esophageal mucosa is sprayed with iodine, allowing for directed biopsy. Several studies have shown that important differences in histologic and cytologic diagnosis and grading exist between Western pathologists and those in China or Japan. Despite these findings, the risk of developing invasive SCC is closely correlated to the severity of ESD encountered. Correct recognition of ESD thus is important. PMID- 11936263 TI - Gastric epithelial dysplasia. AB - Gastric dysplasia is considered to be the penultimate stage of the gastric carcinogenesis sequence. Its clinical importance has been underscored since its association with gastric adenocarcinoma was established. High-grade dysplasia and to a lesser degree low-grade dysplasia are markers of increased cancer risk, although their natural histories are difficult to determine. There are many pitfalls in the diagnosis of gastric dysplasia. Its recognition and grading are subject to interobserver variability, particularly at the lower end of the histologic spectrum (negative v indefinite for dysplasia v low-grade dysplasia) when inflammation is present. Also, diagnostic criteria and grading schemes have evolved differently worldwide resulting in disagreement between pathologists. Against this background, the authors review contemporary issues related to gastric dysplasia, its definition, classification, grading, and natural history. We also discuss new classifications of gastric epithelial dysplasia designed to develop equivalence between grading schemes worldwide. PMID- 11936264 TI - Dysplasia in inflammatory bowel disease. AB - The risk of neoplasia in ulcerative colitis and Crohns colitis increases with both the duration and the extent of disease. In patients with extensive or pancolitis, the cancer risk increases dramatically 8 to 10 years after the first onset of disease. Childhood onset of colitis and primary sclerosing cholangitis further increase the risk of developing colorectal carcinoma. The performance of surveillance endoscopy to identify dysplastic precursor lesions via endoscopic biopsy specimens has become the main management strategy to combat this risk. Biopsies should be classified as negative for dysplasia, indefinite for dysplasia, low-grade dysplasia, or high-grade dysplasia according to standard criteria. A prophylactic colectomy is the procedure of choice when high-grade dysplasia or low-grade dysplasia associated with a lesion or mass is present. Some centers also recommend a colectomy for the presence of low-grade dysplasia in flat mucosa. Given these management recommendations, care should be taken not to overcall reactive epithelial changes in the face of active colitis. All diagnoses of dysplasia should be confirmed, preferably by a pathologist experienced in interpreting gastrointestinal biopsies. PMID- 11936265 TI - Cytologic diagnosis of dysplasia in the alimentary tract. AB - Diagnosis of dysplasia in the gastrointestinal tract has important clinical implications for management decisions. Although dysplasia is generally recognized by histology, cytologic specimens have been shown to increase the sensitivity of tissue diagnosis. The advantage of cytology is the ability to sample a much larger mucosal area. The relatively low specificity caused by the overlap in cytologic features of reactive atypia, dysplasia, and carcinoma is a disadvantage. This article reviews the cytology of squamous and glandular dysplasia in the alimentary tract as well as the studies that have evaluated the clinical utility of cytologic sampling of the gastrointestinal tract in the evaluation of dysplasia. PMID- 11936266 TI - The molecular basis of dysplasia. AB - Detailed descriptions of the molecular pathways involved in dysplasia and carcinoma would fill many bound volumes. This article illustrates the molecular concepts involved in the pathogenesis of dysplasia and carcinoma, with examples that are most relevant to the gastrointestinal tract. At this time, there are at least 4 circuits that must be altered for a cell to become dysplastic: 1) requirement of a continuous or autonomous growth stimulus, 2) development of resistance to growth inhibitory signals, 3) avoidance of apoptosis, and 4) ability to transcend the limits imposed by the generational clock. Although it is likely that 1 or 2 additional circuits must also be altered for the development of carcinoma, they are not defined at this time. PMID- 11936267 TI - Cloning and molecular characterization of a SDS-activated tyrosinase from Marinomonas mediterranea. AB - The sequence of the tyrosinase gene cloned from Marinomonas mediterranea is reported. It is the second tyrosinase cloned from a Gram negative bacterium. Its size is higher than that of Gram positive tyrosinases from Streptomyces, and more similar to the eukaryotic enzymes. Its sequence shares the features of copper binding sites found in all tyrosinases. Based in the comparison of tyrosinases from all types of organisms, an extension of the characteristic signatures existing at Prosite is proposed. This tyrosinase shares with some plant and amphibian tyrosinases a strong specific activation by submicellar concentrations of SDS. Intrinsic fluorescence and kinetic properties indicate that the activation is caused by an SDS-dependent conformational change that facilitates the substrate accessibility to the dicopper active site. PMID- 11936268 TI - Ethnic variation in melanin content and composition in photoexposed and photoprotected human skin. AB - We have examined the quantity and composition of melanin in both photoprotected (volar upper arm) and chronically photoexposed (dorsal forearm) skin from a range of different ethnic skin types including African, Indian, Mexican, Chinese and European. The most lightly pigmented (European, Chinese and Mexican) skin types have approximately half as much epidermal melanin as the most darkly pigmented (African and Indian) skin types. However, the composition of melanin in these lighter skin types is comparatively more enriched with lightly coloured, alkali soluble melanin components (up to three-fold). Regardless of ethnicity, epidermal melanin content is significantly greater in chronically photoexposed skin than it is in corresponding photoprotected skin (up to two-fold). However, by comparison there is only a modest enrichment of lightly coloured, alkali soluble melanin components in photoprotected skin (up to 1.3-fold). Analysis of melanosomes extracted from the epidermis in these subjects indicates that the proportion of spheroidal melanosomes is low in all skin types examined (<10%). This suggests that in human skin, pheomelanin is a very minor component of epidermal melanin, even in the lightest (European) skin types. Analysis of melanosome size revealed a significant and progressive variation in size with ethnicity: African skin having the largest melanosomes followed in turn by Indian, Mexican, Chinese and European. On the basis of these findings, we propose that variation in skin pigmentation is strongly influenced by both the amount and the composition (or colour) of the melanin in the epidermis. Variation in melanosome size may also play a significant role. However, the data also suggest that in human skin there are subtle differences in the mechanisms associated with the maintenance of constitutive pigmentation and facultative hyperpigmentation, respectively. PMID- 11936269 TI - The impact of epidermal melanin on objective measurements of human skin colour. AB - Objective measurements of human skin colour were made with a tristimulus (L*a*b*) chromameter in a range of different ethnic skin types. These were compared with biochemical measurements of melanin content, melanin composition and melanosome size in skin biopsies obtained from the same sites. L*, a* and b* values were found to vary significantly with ethnicity. In general, constitutively dark skin types have lower L* values, higher a* values and higher b* values than constitutively light skin types. Total epidermal melanin content appears to be the primary determinant of L* values in human skin (r = -0.88; P < 0.00001), whilst melanosome size also has a significant but more subtle influence on L* values (r = -0.73; P < 0.00001). There is also a strong positive contribution to a* values from epidermal melanin (r = 0.66, P < 0.00001), which accounts for the ethnic variation in a* values observed in this study. Melanin is also a major contributor to b* values in lighter skin types (r = 0.71, P < 0.00001). However, this relationship breaks down in darker skin types where b* values actually reach a maximum and then decrease as the concentration of melanin in the skin increases. This appears to be because of optical masking of yellow light by high concentrations of melanin in the epidermis. Analysis of the relationships between L*, a* and b* values in human skin indicate that they are very closely interrelated, and suggest that the optical properties of melanin in the epidermis are very similar to those of a dye on a fabric substrate. PMID- 11936270 TI - Melanosomal proteins--role in melanin polymerization. AB - Melanin, the major determinant of skin colour, is a tyrosine-based heteropolymer of indeterminate molecular weight. In vivo, melanin synthesis occurs within highly specialized organelles called melanosomes. Coated vesicles encapsulating the enzyme tyrosinase and tyrosinase related proteins, fuse with premelanosomes that contain structural proteins to form mature melanosomes. Coated vesicles and premelanosomes have been shown to have only melanin monomers but not the polymer. Our earlier results have clearly shown that the presence of proteins other than tyrosinase are critical for the post-tyrosinase steps of melanin polymerization at acidic pH. Proteins in melanosomes are difficult to purify because of their firm association with melanin. Thus, with progressive melanization, melanoproteins become progressively insoluble. In this paper, we discuss the isolation and purification of melanosomal proteins and their role in melanin polymerization. We have hypothesized that the initiation of polymerization and the binding of melanin to proteins are two discrete events and we have developed assays to quantify these events. Purified melanosomal proteins differ in their ability to polymerize melanin monomers. Further, we have also shown that two polypeptides (28 and 45 kDa) purified from melanosomes inhibit melanin polymerization but can bind preformed melanin. In conclusion, melanosomal proteins regulate melanin polymerization and differ in their ability to bind melanin. Polymerization and binding abilities of melanosomal proteins are specific to each protein and melanin-protein interaction is not nonspecific. PMID- 11936271 TI - Vitiligo: cytomegalovirus associated? PMID- 11936272 TI - Du temps perdu a la recherche. AB - An attempt has been made to justify an academic career by outlining the approximate history of the development of ideas and tracing their origins and arborization. Because there are multifarious influences and formative factors involved in their genesis it has proved almost impossible to acknowledge all the important contributors to the patchwork of investigations that are summarized. However, whilst the review is incomplete and necessarily brief, it is hoped that a glimpse of some twentieth century preoccupations with biological science is afforded. PMID- 11936273 TI - The pallidin (Pldn) gene and the role of SNARE proteins in melanosome biogenesis. AB - This review focuses on the product of the pallidin (Pldn) gene, one of a number of genes that in mice are associated with pigmentation defects and platelet dense granule deficiency. A similar combination of defects is also observed in patients suffering from Hermansky-Pudlak (HPS) and Chediak-Higashi (CHS) syndromes. Pldn encodes a novel, approximately 20-kDa protein that is expressed ubiquitously in mammalian tissues. The pallidin protein was found to bind to syntaxin 13, a member of the syntaxin family of soluble N-ethylmaleimide-sensitive factor attachment protein receptors (SNAREs). As SNARE proteins mediate fusion of intracellular membranes, pallidin may play a role in membrane fusion events required for melanosome biogenesis. PMID- 11936274 TI - New insights into the pathogenesis of vitiligo: imbalance of epidermal cytokines at sites of lesions. AB - Vitiligo is a skin disease that is caused by selective destruction of melanocytes and is characterized by white spots. Melanocytes and keratinocytes seem to exhibit a functional close relationship, mediated at least in part by keratinocyte-derived cytokines, which seem important for survival and activity of melanocytic cells. We wanted to investigate the hypothesis that in vitiligo the expression of epidermal cytokines may be modified compared with normal skin. In 15 patients with active, non-segmental vitiligo, biopsies were obtained from lesional, perilesional and non-lesional skin; normal skin from five healthy donors was also tested. Tissue sections were tested using immunohistochemistry for the expression of keratinocyte-derived cytokines with stimulating activity, such as granulocyte-monocyte colony stimulating factor (GM-CSF), basic fibroblastic growth factor (bFGF), and stem cell factor (SCF) or with inhibiting activity, such as interleukin 6 (IL-6) and tumour necrosis factor alpha (TNF alpha) on melanocytes. Cytokine receptors and specific melanocytic markers were also investigated. No melanocyte was identified in lesional skin by means of specific markers or c-kit receptor, whereas in perilesional, non-lesional and healthy skin, melanocytes were found in similar number. In vitiligo skin a significantly lower expression of GM-CSF, bFGF and SCF was found, and a significantly higher expression of IL-6 and TNF-alpha was detected, compared with perilesional, non-lesional and healthy skin. In conclusion, we provided evidence that a significant change of epidermal cytokines exists in vitiligo skin compared with perilesional, non-lesional and healthy skin, suggesting that the cytokine production of epidermal microenvironment may be involved in vitiligo. PMID- 11936275 TI - 1H NMR spectroscopic characterization of solutions of Sepia melanin, Sepia melanin free acid and human hair melanin. AB - A 1H nuclear magnetic resonance study of Sepia melanin, Sepia melanin free acid (Sepia MFA) and human hair melanin was carried out in deuterium oxide solution at pH 10-11. The empirical formula of Sepia MFA was calculated and used to estimate the number of protons in the aromatic region of the Sepia MFA polymeric chain and to suggest a possible monomeric unit profile. PMID- 11936276 TI - The expression of corticotropin-releasing hormone in melanoma. AB - We previously demonstrated that advanced melanoma cells express high amounts of proopiomelanocortin (POMC) that correlate with tumor progression. We now investigated whether the high expression of POMC derives from increased expression of corticotropin-releasing hormone (CRH) and the possible role of CRH as a melanoma growth factor. Forty-five cases of melanoma [25 primary malignant melanoma; 20 metastatic melanoma (MetM)] were immunohistochemically analysed for coexpression of POMC and CRH peptides. The ability of CRH to induce POMC expression in cultured melanoma cells was examined using CRH and a CRH antagonist. In CRH positive melanomas, seven out of nine cases (78%) of primary melanoma, and 7 out of 12 cases (58%) of MetM showed colocalization of CRH and POMC peptides. CRH induced POMC mRNA expression, an effect that was inhibited by a CRH antagonist. These results provide evidence for the existence of the CRH/POMC axis in pigmented lesions. PMID- 11936277 TI - Dynamics of recovery of morphometrical variables and pQCT-derived cortical bone properties after a short-term protein restriction in maturing rats. AB - Severe protein restriction during the post-weaning period in the rat markedly reduces femoral bone mass and produces a number of alterations in the shaft biomechanical properties. Body weight and femur length show an immediate and complete catch-up during nutritional rehabilitation. The aim of the present investigation was to assess whether the accelerated bone growth that occurs during protein rehabilitation is accompanied by recovery of cortical bone properties. The dynamics of the recovery of both material and geometric properties were thus evaluated on the femoral diaphyses in 45-day old female rats after a 10-day period of dietary protein restriction by peripheral quantitative computed tomography (pQCT). Protein starvation led to marked reduction of both body weight and femoral length (37% and 14% at day 10, respectively) which showed a complete catch-up after 30 d of protein refeeding. Protein restriction was associated with the interruption of the natural increase in cortical area (CtCSA), volumetric cortical bone mineral content (vCtBMC) and volumetric cortical bone mineral density (vCtBMD) which were 19.7, 25.8, and 14%, respectively, in malnourished than in control rats at the end of the protein starvation period. These parameters recovered completely during protein refeeding. Treatment also reduced by 30% both rectangular (xCSMI) and polar (pCSMI) moments of inertia. Although an improvement of these architectural indicators occurred with time, an approximately 20% deficit was still present at the end of the observation period (70 d), as was the bone strength index (BSI). It is concluded that protein restriction affected the adaptation of diaphyseal design which should reduce the mechanical competence of the femoral diaphysis because of an inadequate architectural distribution of cortical bone, and that the alteration did not show complete catch-up during the studied period. PMID- 11936278 TI - Growth promotion in chickens by interleukin-2. AB - Chicken interleukin-2 (cIL-2), which was prepared by sensitizing chicken lymphocytes with concanavalin A, was administered to fertile broiler eggs on Day 18 of embryonation (0.1 mg in 200 mL distilled water). Controls (CON) received distilled water. Hatched chicks were reared to 6 wk. Body weight (BW), as well as abdominal fat pad, liver, bursa of Fabricius, a thymic lobe, spleen, and gonads were excised and expressed relative to BW at 2, 4, and 6 wk of age. Additionally, hematocrit (HCT), hemoglobin (HGB), and plasma protein (PP) levels were determined at the three time intervals. Finally, chicks were sensitized against human gamma globulin (HGG) and challenged at 6 wk by intradermal injections into the wattles. Delayed type hypersensitivity (DTH) to HGG was used as a direct measure of cell-mediated immunity. In ovo cIL-2 increased BW consistently, relative fat pad weights at 2 wk, relative bursa and liver weights at 2 and 6 wk, HBG and relative thymic weight at 2 and 4 wk, and PP at 2 wk. Delayed type hypersensitivity to HGG was not affected by cIL-2. Potential metabolic and immunologic mechanisms to explain in ovo cIL-2 effects are discussed. PMID- 11936279 TI - Relative food intake of rats submitted to a moderate transgenerational undernutrition. AB - An experiment on rat undernutrition through seven generations was performed in order to see: (1) whether the nutritional stress on growth increases from one generation to the next, and (2) if an equilibrium point (AFP) in which the RFI- the amount of food intake (mg) per gram of body weight--reached is the same in both control and undernourished animals. The RFI values were calculated for each generation, between the 30th and 100th days of age. A moderate undernutrition was applied to the seven generations (F1 to F7) following the parental (P) one, which acted as controls. Undernourishment was made from conception to the end of the experiment (100 days old). The RFI values diminished with the age increment and increased through generations. There was, however, a clear AFP of 75.9 +/- 3.5 mg/g at 100 days of age in males, and of 78.7 +/- 4.2 mg/g at 90 days of age in females. A clear cumulative increment of RFI through the filial generations was also found at intermediate growth ages. The frequently argued nongenetic transmission of the nutritional deficiencies from parents to descendants was corroborated with the present results. Such cumulative effect was evident at ages before the AFP was reached; i.e., when the decrement in body mass of the undernourished animals was not yet equilibrated with the amount of available nutrients. PMID- 11936280 TI - Faecal incontinence: common and treatable. PMID- 11936281 TI - Hospital care for Aboriginals and Torres Strait Islanders: appropriateness and decision making. PMID- 11936282 TI - Measles transmission in healthcare settings in Australia. PMID- 11936283 TI - Liver biopsy in the 21st century: where and why? PMID- 11936284 TI - Prevalence of faecal incontinence and associated risk factors; an underdiagnosed problem in the Australian community? AB - OBJECTIVE: To determine the prevalence of faecal incontinence in the community and evaluate identifiable risk factors. DESIGN AND SETTING: Cross-sectional survey using a validated questionnaire. A short version of the questionnaire was sent to 220 subjects and a long version to 770 subjects, randomly selected from western Sydney, Australia. MAIN OUTCOME MEASURES: Self-reported faecal incontinence, defined as involuntary loss of anal sphincteric control leading to unwanted release of liquid or solid faeces (not flatus) at an inappropriate time or in an inappropriate place, within the past 12 months. The long questionnaire also sought information on bowel habit and potential risk factors for faecal incontinence. RESULTS: The response rate was 66%. The prevalence of solid or liquid faecal incontinence was 2% and 9%, respectively. The mean age of subjects with faecal incontinence was 53 years; 55% were women. After adjusting for age and sex, there was a significant association between faecal incontinence and perianal injury (P = 0.03), perianal surgery (P < 0.001), feelings of incomplete defecation (P < 0.0001), loose or watery motions (P < 0.0001) and urgency (P < 0.0001). Seven of 48 subjects with faecal incontinence reported being asked by their physician about faecal incontinence and nine of 33 reported seeking medical advice for their incontinence. Subjects with faecal incontinence perceived their health to be significantly poorer than did other subjects (P = 0.02). CONCLUSION: There is a high burden of faecal incontinence in the community, and the prevalence in men may be greater than is usually appreciated. Despite significant associated morbidity, most cases of faecal incontinence were unrecognised by doctors. PMID- 11936285 TI - Diagnostic and therapeutic procedures among Australian hospital patients identified as indigenous. AB - OBJECTIVES: To determine whether hospital patients identified as Indigenous are less likely than other inpatients to have a principal procedure recorded, and the extent to which any disparity in procedure use can be explained by differences in patient, episode and hospital characteristics. DESIGN: Retrospective analysis of routinely collected administrative data from the National Hospital Morbidity Database (NHMD). SETTING: Australian public and private hospitals. PATIENTS: All patients included in the NHMD whose episode type was recorded as acute and whose separation occurred between 1 July 1997 and 30 June 1998. Patients admitted for routine dialysis treatment were excluded. MAIN OUTCOME MEASURE: Whether a principal procedure was recorded. RESULTS: In public hospitals, patients identified as Indigenous were significantly less likely than other patients to have a principal procedure recorded, even after adjusting for patient, episode and hospital characteristics (adjusted odds ratio [OR], 0.67; 95% CI, 0.66-0.68). This disparity was apparent for most diseases and conditions. In private hospitals, no significant difference was observed (adjusted OR, 0.94; 95% CI, 0.83-1.06). CONCLUSIONS: The disparity in procedure use after adjustment for relevant factors indicates that in Australian public hospitals there may be systematic differences in the treatment of patients identified as Indigenous. PMID- 11936286 TI - Reported management of early-pregnancy bleeding and miscarriage by general practitioners in Victoria. AB - OBJECTIVES: To describe the management of early-pregnancy bleeding and miscarriage reported by general practitioners in Victoria. DESIGN, SETTING, AND PARTICIPANTS: Self-administered, mailed survey of a stratified random sample of GPs in Victoria. Responses weighted by strata to reflect GP population. MAIN OUTCOME MEASURES: Reported management in referral; investigation (especially ultrasound); expectant versus interventional management; and prevention of rhesus iso-immunisation RESULTS: 382 of 621 eligible GPs responded (response rate, 62%). GPs' reported referral was more likely if the patient had painful bleeding (55%) or if the pregnancy was not viable (77%). Ultrasound strongly influenced the assessment of bleeding. Two-thirds of doctors (262/369; 66%) would routinely order ultrasound for painless bleeding, and 328/369 (84%) for painful bleeding. Expectant management was recommended by 15/353 (4%) for incomplete miscarriage with light bleeding and by 6/351 (2%) when bleeding was heavy. Some GPs are uncertain of the indications for anti-D prophylaxis, including instrumentation of the uterus, for which 261/337 (77%) said they would routinely offer anti-D. There was less agreement about anti-D after threatened miscarriage, for which 213/353 (57%) said they offered the injection. CONCLUSIONS: GPs need a working knowledge of the management of early-pregnancy bleeding, and can probably encourage more rational management. There are significant areas where GPs are uncertain, often reflecting uncertainty elsewhere, and some areas where a minority of GPs are not aware of essential requirements. PMID- 11936288 TI - Sickle-cell/beta+-thalassaemia in a Papua New Guinean: the first reported case of the sickle gene in Papua New Guinea. AB - We document the first case of the sickle-cell gene--Hb-S gene importation leading to Hb-S/beta-thalassaemia double heterozygosity--in the apparently previously Hb S gene-free setting of Papua New Guinea. PMID- 11936287 TI - Short-stay, out-of-hospital, radiologically guided liver biopsy. AB - OBJECTIVE: To evaluate the safety, the quality and adequacy of specimens obtained and the cost benefits associated with performing liver biopsy out of hospital, on a short-stay basis, using radiological guidance. DESIGN AND SETTING: A prospective study undertaken over a three-year period, from March 1998 to March 2001, in a private radiology practice. PATIENTS AND PROCEDURES: 251 patients (159 men) with stable liver disease participated. Coagulation studies were performed within a two-week period before biopsy, which was carried out under the guidance of ultrasound (143 patients) or computed tomography (108 patients). A disposable, spring-loaded gun with an 18-gauge biopsy needle was used in each case. A repeat ultrasound or CT scan was performed after the procedure to monitor for complications such as haemorrhage. MAIN OUTCOME MEASURES: Complications of liver biopsy; adequacy of specimens for histological examination; cost of out-of hospital procedures compared with liver biopsies performed in the hospital setting. RESULTS: Two hundred and twenty nine patients (91.2%) were discharged 60 minutes after the biopsy. The only post-biopsy complication was pain, either at the biopsy site or in the right shoulder. Pain was severe in three patients and, for one of these patients, a subcapsular hepatic haematoma was found on ultrasound eight days after the biopsy. Sufficient material for histopathological examination was obtained from all patients. The cost of out-of-hospital biopsies was substantially less than the cost of hospital-based, day-stay procedures. CONCLUSIONS: Short-stay, out-of-hospital, radiologically guided liver biopsy is safe for patients who have stable chronic liver disease and acceptable coagulation profiles. PMID- 11936289 TI - Breaking the deadlock over an Australian trial of injectable opioid maintenance. PMID- 11936290 TI - What's wrong with the dead body? Use of the human cadaver in medical education. AB - A 2000 survey of Australian medical schools showed that use of anatomical dissection and autopsies for teaching has declined. Opinions vary between schools as to the effect on teaching of anatomy and pathology. However, exposure to the dissected human body may have benefits beyond this teaching, including inculcating the trait of "detached concern", teaching about medical fallibility and uncertainty, and raising issues of death and dying. PMID- 11936291 TI - Reclaiming the lost meanings of medicine. PMID- 11936292 TI - End-of-life issues: case 2. AB - When a dying patient lacks decision-making capacity, the general practitioner needs to collaborate with family members in making decisions about forgoing life sustaining treatment. The key to working out the best course of action is for the doctor to have a very clear idea of which treatment options he or she considers acceptable or unacceptable. The choice of treatment depends on a thorough evaluation of all the clinical information and careful reflection, bearing in mind that medicine has its own proper limits. Life-sustaining treatment may legitimately be forgone if it is (a) therapeutically futile, (b) overly burdensome to the patient, (c) not reasonably available without disproportionate hardship to the patient's carers or others, or (d) refused by the patient. PMID- 11936293 TI - Is dexamethasone effective in treating acute migraine headache? PMID- 11936294 TI - Asylum seekers and healthcare. PMID- 11936295 TI - Asylum seekers and healthcare. PMID- 11936296 TI - Asylum seekers and healthcare. PMID- 11936297 TI - COX-2 inhibition and thrombotic tendency. PMID- 11936298 TI - Evidence-based healthcare 10 years on: is the National Institute of Clinical Studies the answer? PMID- 11936299 TI - Liver biopsy in hepatitis C: reassessing its role in 2001. PMID- 11936301 TI - The decline in hospital autopsy rates in 2001. PMID- 11936300 TI - Changes in serum folate concentrations following voluntary food fortification in Australia. PMID- 11936302 TI - How much alcohol is drunk in Australia in excess of the new Australian alcohol guidelines? PMID- 11936303 TI - Telephone triage in Western Australia. AB - We describe the development and operations of the first large-scale Australian medical telephone triage centre. Studies have commenced to evaluate efficacy and safety of the service, as well as gauge the impact on demand for healthcare services. PMID- 11936304 TI - Diabetes detection in Australian general practice: a comparison of diagnostic criteria. AB - OBJECTIVES: To study the influence of different diagnostic criteria on the prevalence of diabetes mellitus and characteristics of those diagnosed. DESIGN AND SETTING: Retrospective analysis of data from the general-practice-based Australian Diabetes Screening Study (January 1994 to June 1995). PARTICIPANTS: 5911 people with no previous diagnosis of diabetes, two or more symptoms or risk factors for diabetes, a random venous plasma glucose (PG) level > 5.5 mmol/L and a subsequent oral glucose tolerance test (OGTT) result. MAIN OUTCOME MEASURE: Prevalence of undiagnosed diabetes based on each of three sets of criteria: 1997 criteria of the American Diabetes Association (ADA), 1996 two-step screening strategy of the Australian Diabetes Society (ADS) (modified according to ADA recommendations about lowered diagnostic fasting PG level), and 1999 definition of the World Health Organization (WHO). RESULTS: Prevalence estimates for undiagnosed diabetes using the American (ADA), Australian (ADS) and WHO criteria (95% CI) were 9.4% (8.7%-10.1%), 16.0% (15.3%-16.7%) and 18.1% (17.1%-19.1%), respectively. People diagnosed with diabetes by fasting PG level (common to all sets of criteria) were more likely to be male and younger than those diagnosed only by 2 h glucose challenge PG level (Australian and WHO criteria only). The Australian (ADS) stepwise screening strategy detected 88% of those who met the WHO criteria for diabetes, including about three-quarters of those with isolated post-challenge hyperglycaemia. CONCLUSION: The WHO criteria (which include an OGTT result) are preferable to the American (ADA) criteria (which rely totally on fasting PG level), as the latter underestimated the prevalence of undiagnosed diabetes by almost a half. The Australian (ADS) strategy identified most of those diagnosed with diabetes by WHO criteria. PMID- 11936305 TI - Menstrual and contraceptive management in women with an intellectual disability. AB - OBJECTIVE: To review the clinical management of young women with intellectual disabilities with menstrual and contraceptive concerns. DESIGN: Prospective cohort study of all girls and young women with a significant intellectual disability and moderate to high support needs who presented at my gynaecology clinic for management of menstrual and contraception-related issues in the period 1990-1999. SETTING: Gynaecology clinic at the Centre for Adolescent Health, Royal Children's Hospital, Melbourne, and my private consulting rooms. OUTCOME MEASURES: The clinical management options considered most appropriate for these women, including advice, reassurance, medication (oral contraceptive pill, non steroidal anti-inflammatory drugs, depomedroxyprogesterone acetate, hormone replacement therapy) and surgical options. RESULTS: For 2 of 107 young women, surgical approaches were required to manage their menstrual problems or contraception-related issues. For the remainder of the women, information, advice or medical management were sufficient. CONCLUSIONS: Management of the menstrual and contraceptive needs of young women with an intellectual disability is similar in most cases to the management of non-disabled women. Surgical management is required infrequently. PMID- 11936306 TI - Emergency endovascular stent-graft repair for thoracic aortic injury. AB - We report a case of emergency endovascular stent-graft repair of a traumatic false aneurysm of the thoracic aorta. Thoracotomy was relatively contraindicated because the patient also sustained incomplete spinal injury, He recovered fully from both chest and spinal injuries, and remains without evidence of complications related to the stent-graft 18 months after the injury. PMID- 11936307 TI - Sharp v Port Kembla RSL Club: establishing causation of laryngeal cancer by environmental tobacco smoke. AB - A New South Wales Supreme Court jury has decided that environmental tobacco smoke (ETS) can cause or materially contribute to the development of laryngeal cancer. Evidence presented that ETS may cause or materially contribute to laryngeal cancer included the molecular genetics of tobacco-smoke-induced carcinogenesis, and two relevant epidemiological studies. The plaintiff's exposure to ETS was established indirectly, on the basis of occupational history involving work as a bar attendant in licensed premises. The jury's decision seems likely to encourage other "passive smoking" cases, and may result in measures to reduce occupational exposure to ETS. PMID- 11936308 TI - Ethical issues concerning the relationships between medical practitioners and the pharmaceutical industry. AB - Medical practitioners and the pharmaceutical industry serve interests that sometimes overlap and sometimes conflict. There is strong evidence that associations between industry and doctors influence the behaviour of the latter in relation to both clinical decision making and the conduct of research. In view of the risk of compromising relationships with patients and the integrity of the research process, doctors must exercise care in their dealings with industry. The basic principles underlying the conduct of doctors with respect to pharmaceutical companies should be openness and transparency. Clearly articulated procedures should be developed to deal with specific issues such as travel subsidies, receipt of gifts, sponsorship of conferences and continuing education activities, and dualities of interest arising in clinical and research settings. PMID- 11936309 TI - Declarations made by graduating medical students in Australia and New Zealand. AB - OBJECTIVE: To survey the use of declarations of ethical commitment made by graduating medical students in Australia and New Zealand. METHODS: Information, obtained by email and telephone, from faculty officers of all faculties of medicine (or medicine and health sciences) in Australia and New Zealand. RESULTS: Declarations are made by graduating medical students at seven of 12 Australasian faculties of medicine. To date, declarations have been based on the Declaration of Geneva or the Hippocratic Oath or have been formulated by academic staff or the students themselves. In six of the seven universities, declarations are made as part of a special declaration ceremony (usually combined with a prize-giving ceremony). One university includes a declaration as part of the official graduation ceremony. DISCUSSION: We discuss the relative merits of a declaration selected for students by staff members and a declaration written anew by each group of graduating students. PMID- 11936310 TI - Surgical management of amiodarone-associated thyrotoxicosis. PMID- 11936311 TI - Tepid sponging and paracetamol for reduction of body temperature in febrile children. PMID- 11936312 TI - Is bupropion (Zyban) causing deaths? PMID- 11936313 TI - Carotid stenting or endarterectomy for stroke prevention. PMID- 11936314 TI - Carotid stenting--current caution. PMID- 11936315 TI - Content of isoflavone-containing preparations. PMID- 11936316 TI - Screening for gestational diabetes: the time of day is important. PMID- 11936317 TI - Changing demographics of cervical carcinoma. PMID- 11936318 TI - Ethics and evidence-based medicine. PMID- 11936319 TI - Ethics and evidence-based medicine. PMID- 11936320 TI - Serum alanine aminotransferase levels and the detection of hepatitis C virus (HCV) in chronic HCV infections. PMID- 11936321 TI - Prejudice against mental illness. PMID- 11936322 TI - Nurse-led telephone advice. PMID- 11936323 TI - When is diabetes really diabetes? PMID- 11936324 TI - Medical commencement oaths: shards of a fractured myth, or seeds of hope against a dispiriting future? PMID- 11936325 TI - Health services at breaking point. ...don't panic? PMID- 11936326 TI - Antibiotic resistance in uncomplicated urinary tract infection: problems with interpreting cumulative resistance rates from local community laboratories. AB - AIMS: To determine the resistance rates and patterns in bacteria causing uncomplicated urinary tract infections (UTIs) presenting to general practitioners (GPs) in Christchurch. METHODS: 82 randomly selected GPs in Christchurch participated in the study. Midstream urine (MSU) samples were prospectively collected for standard microbiological analysis on all women between the ages of 16 and 50 years presenting with symptoms of dysuria and frequency and who had positive dipstick testing to either (or both) nitrites or leucocytes. MSUs were submitted for bacterial colony counts and resistance testing of isolates present in adequate numbers. RESULTS: 374 specimens were collected. 299 filled the inclusion criteria, of which 94 fulfilled criteria for significant infection. Trimethoprim resistance was found in 8, (8.5%) (95%CI 2.8,14.2) overall with a resistance rate for Escberichia coli (E. coli) to trimethoprim of 11.5%. This compared with cumulative resistance rates from local community laboratories for E. coli to trimethoprim of 19%. For a woman in this age group presenting with symptoms of UTI we estimated that her probability of having a trimethoprim resistant organism was 2.7%. CONCLUSION: Trimethoprim remains a reasonable first line treatment for uncomplicated UTI in Christchurch. Actual resistance rates are significantly less than those derived from routine pooled laboratory specimens, and when used in an intention to treat calculation to inform empiric prescribing, become even less significant. While collection of these routine data is essential to provide early warning of emergent resistance, a truly representative rate should be determined to inform prescribing decisions if resistance appears to be increasing. PMID- 11936327 TI - Heart failure: ethnic disparities in morbidity and mortality in New Zealand. AB - AIMS: To compare heart failure outcomes for Maori and non-Maori New Zealanders. METHODS: Restrospective analysis of New Zealand mortality (1988-97) and hospital admissions (1989-98) due to heart failure for Maori and non-Maori, aged 45 years and over. RESULTS: Mortality from heart failure was more than 8.8 times higher among Maori males aged 45-64 years than non-Maori (95% confidence interval 7.6 to 10.1), and 3.5 (3.1 to 4.1) times higher among Maori aged 65 years and over. Mortality ratios for females were similar. Hospital admissions with a primary diagnosis of heart failure averaged eight times higher among Maori males aged 45 64 and nine times higher among Maori females compared to non-Maori. Maori males and females aged 65 years and over had three and four times the non-Maori rate of admission. CONCLUSIONS: Disparities between Maori and non-Maori in outcomes for heart failure are high in New Zealand. Effective, evidence-based interventions have not yet impacted on populations most 'at risk'. PMID- 11936328 TI - False positive 131I-metaiodobenzylguanidine scan in the postoperative assessment of malignant phaeochromocytoma secondary to medullary hyperplasia. PMID- 11936329 TI - Hepatitis B--prevention of perinatal transmission. PMID- 11936330 TI - Doing our best for patients with stroke. PMID- 11936331 TI - "But it's mine--isn't it?". PMID- 11936332 TI - Patient requests for unwarranted treatment. PMID- 11936333 TI - Antenatal screening for HIV. PMID- 11936334 TI - Ethics committees and research. PMID- 11936335 TI - No fault compensation for medical errors: who wins? PMID- 11936336 TI - No fault compensation for medical errors: who wins? PMID- 11936337 TI - Median nerve injury mimicking carpal tunnel syndrome. PMID- 11936338 TI - Audit of cervical screening, the law and ethics committees. PMID- 11936339 TI - Perinatal database. PMID- 11936340 TI - Public health surveillance needs and cancer registration data. PMID- 11936341 TI - Are Maori under-served for cardiac intervention? PMID- 11936342 TI - Creeping privatisation? PMID- 11936344 TI - Self care and family care. PMID- 11936345 TI - Comparison of radiation doses for screening mammography and chest CT for coronary artery calcification. PMID- 11936346 TI - Inappropriate prescribing. PMID- 11936347 TI - Guilty of conduct unbecoming. PMID- 11936348 TI - Acute stroke services in New Zealand. AB - AIMS: To obtain an overall picture of the organisation of acute stroke management in hospitals throughout New Zealand. METHODS: A questionnaire was sent to all New Zealand hospitals. The survey included questions about access to organised stroke care, the presence of designated areas for stroke patient management, guidelines for stroke management and audit. RESULTS: Responses were received from all hospitals surveyed, with 41 admitting stroke patients acutely. Five hospitals (four regional and one large urban) had organised inpatient care. Five hospitals (three regional and two large urban) had stroke physicians. Only 40-60% of the New Zealand population had access to hospitals with guidelines for the management of complications following stroke or secondary prevention. Only fifteen of 41 hospitals had audited local stroke care. There were few differences in the management of stroke patients between urban and regional centres, but care in some regional hospitals was 'better' than that in most large urban hospitals. CONCLUSIONS: The development of an organised approach to inpatient stroke care in New Zealand and the training of general physcians, geriatricans and neurologists in stroke medicine must be seen as a priority. PMID- 11936349 TI - Time trends and seasonal patterns of asthma deaths and hospitalisations among Maori and non-Maori. AB - AIM: To examine the time trends and seasonal patterns in asthma mortality and hospitalisations in Maori and non-Maori. METHODS: We studied asthma deaths in Maori and non-Maori during 1962-1998 and asthma hospitalisations during 1976-1998 in the 5-34 and 35-74 year age-groups. Average monthly mortality and hospital discharge rates were calculated for 1978-1998 to observe the seasonal patterns. RESULTS: The two asthma mortality epidemics of the 1960s and 1970s affected Maori disproportionately, with the peak rates in 1979 being twice that of non-Maori (7.4 vs 3.7 per 100 000) although the rates are now similar. Hospitalisation rates for Maori were consistently higher than non-Maori with the rates remaining elevated until 1987. The most recent hospitalisation figures show a fall of 61.2% from the 1987 rate. For Maori in the 5-34 year age-group, deaths were highest in spring while hospitalisations peaked in the winter month of May. CONCLUSION: Although asthma mortality and hospitalisation rates for Maori have declined over the period under study, Maori continue to experience more severe asthma and have a higher number of hospital admissions for asthma than non-Maori. PMID- 11936350 TI - Changing the minimum legal drinking age--its effect on a central city emergency department. AB - AIMS: To quantify the effect of a recent national law change on the presentation of ethanol intoxicated patients to a central city Emergency Department (ED). METHODS: All records of ethanol intoxicated patients presenting to the ED for twelve months before and after the change to the minimum legal drinking age were studied. Each patient was classified as having laboratory confirmed intoxication, clinical suspicion only, or no record of intoxication. Three age groups were identified, 15-17 year olds, eighteen and nineteen year olds and over 20 year olds. Within each age group the proportion of presentations with ethanol intoxication was compared across the two time periods. RESULTS: The number of intoxicated 18 and 19 year olds increased in the twelve months after the national law change from 66 to 107 (52 to 80 for laboratory confirmed intoxication and fourteen to 27 for clinical suspicion only). This represented an increase in the proportion of presentations in this age group with intoxication (p=0.009) from 2.9% to 4.4%, a 50% increase (RR=1.51, 95%CI 1.11-2.03). There was no evidence of an increase in the proportion intoxicated for those over nineteen years (3.4% vs 3.3%, p=0.48, RR=0.97, 95%CI=0.89-1.06) although the numbers increased slightly (963 to 992). However there was a worrying trend for an increase in the 15-17 year olds, with numbers increasing from 72 to 95 and the proportion increasing from 5.0% to 6.7% (p=0.07, RR=1.35, 95%CI=0.98-1.88). CONCLUSION: The recent lowering of the minimum legal drinking age from 20 to eighteen years has resulted in increased presentations to the ED of intoxicated eighteen and nineteen year olds. A similar trend was seen in the 15-17 year olds. PMID- 11936351 TI - Antibiotic prophylaxis in endoscopic and minimally invasive surgery. AB - Infections are unusual following minimally invasive surgery but antibiotic prophylaxis is given in the same way as for the open surgery equivalents. Most prophylactic regimens have not been subjected to randomised placebo controlled trials. Antibiotic prophylaxis has been shown to be beneficial in transurethral resection of the prostate. In endoscopic retrograde cholangiopancreatography (ERCP) the incidence of bacteremia, but not cholangitis, is reduced by prophylaxis and some do not recommend its routine use. For arthroscopies and laparoscopies infection is too rare to warrant prophylaxis. There is a theoretical risk of infection in that endoscopes cannot withstand autoclaving so only high level disinfection can be used between patients. However, for most minimally invasive operations, the small wound size, reduced immune challenge and rapid recovery of the patient outweigh any disadvantages compared with open surgery. PMID- 11936352 TI - Antibiotic prophylaxis in clean surgery: breast surgery and hernia repair. AB - Use of prophylactic antibiotics in clean surgery is still controversial. We reviewed the literature of the last 10 years to identify the best way to approach clean surgery. The question is more important for patients undergoing breast surgery. The presence of an infected breast wound delays the beginning of postoperative adjuvant anticancer therapy: there is good evidence to suggest that delayed adjuvant therapy compromises the outcome for patients in terms of both local control and survival. There are several clinical trials that have addressed the efficacy of prophylactic antibiotics for patients undergoing breast surgery and hernia repair. Platt et al assessed the efficacy of preoperative antibiotic prophylaxis in a clinical trial of 1218 patients undergoing clean surgery with an absolute reduction rate of 39% in wound infections. Gupta et al reported no influence on the incidence of infective complications by antibiotic prophylaxis in 357 patients undergoing elective breast surgery. Like breast surgery, use of prophylaxis in hernia repair is not clear: a prospective, randomized, double blind, multicenter study of 619 patients assessed no benefit of antibiotic prophylaxis. On the other hand Lewis et al reported a 75% reduction of infections in low-risk patients when a single dose of cefotaxime was used in clean operations. A particularly interesting point is the use of prosthetic mesh in hernia repair and primary reconstructive surgery in breast surgery. Amland et al reported a significant reduction of the incidence of wound infections in a group of patients undergoing reconstructive breast surgery, receiving azithromycin vs placebo (5% vs 20%). In hernia repair we stress the need to prevent wound infections: currently Liechtestein's technique is widely performed all over the world. Mesh infection is an unpleasant event that requires prosthesis removal. The lack of conclusive studies about antibiotic prophylaxis in clean surgery suggests that a single-dose of cephalosporin at the induction of anesthesia may be prudent. This procedure is certainly inexpensive and safe and, more importantly, probably does not have an impact on antibiotic resistance. PMID- 11936353 TI - Role of glycopeptides in the treatment of septic complications after cardiac surgery. AB - Agents like Staphylococcus epidermidis and Staphylococcus aureus are common agents in both early and late prosthetic valve endocarditis (PVE). Streptococci, especially vividans and enterococci are more apt to occur late. Diphtheroids and gram-negative bacteria are also frequent in early and late PVE. Fungi are found at a frequency of 5 to 8% and a variety of unusual organisms are found in individual case reports. Treatment is based on parenteral therapy with a bactericidal agent that can achieve trough serum levels in excess of 8-10 X MICs of the infecting organisms. Initially the antibiotic selection should be active against the most common isolates. Because most S. epidermidis are beta-lactam resistant, vancomcyin must be part of the initial empiric regimen. Vancomycin should be combined with rifampin or an aminoglycoside (usually gentamicin) or both. When there is a high level of resistance to aminoglycoside, vancomycin may be used alone until susceptibility data are available and then rifampin can be given together with an aminoglycoside or a quinolone to which the organism is susceptible. The aminoglycoside should be given for a maximum of 2 weeks, to avoid nephrotoxicity, and vancomycin for 6 weeks. Surgery is required in case of major emboli, hemodynamic decompensation, and uncontrolled infection. The presence of bacteremia for more than 1 week may warrant surgical intervention but, if the patient appears to be well and without emboli or hemodynamic problems, serum levels of antibiotic, particularly vancomycin, should be evaluated. Dosage regimen should be modified to achieve trough levels of vancomycin between 15 to 20 microg/ml. Use of vancomycin by continuous infusion may be considered with a targeted blood concentration of 15 to 20 microg/ml. PMID- 11936354 TI - Antibiotic prophylaxis in clean neurosurgery. AB - Two regimens of antibiotic prophylaxis are in use at our institution. These protocols consist in perioperative administration of a single dose of amoxicillin clavulanic acid of 2.2 g at induction and 8 h later and irrigation of the surgical wound with rifamycin before closure. In cases of dirty surgery, placement of external shunts or foreign bodies, we administer vancomycin 1.5 g/die and ceftazidime 6 g/die for 72 h. A retrospective study of all the clean operations we performed in the last 2 years yielded a total of 793 consecutive procedures with three postoperative wound infections. These results are in agreement with the majority of series reported in literature, although different prophylactic protocols are applied and in some cases no prophylactic antibiotics are administered at all. The use of these agents in clean neurosurgery remains, as a matter of fact, controversial. In order to further investigate this issue we took three or more intraoperative samples for culture in 40 clean cases. Only in 2% of these samples were cultures positive. Although lacking statistical significance, these results are interesting indications of the appropriateness of antimicrobial prophylaxis in clean neurosurgery and invite further investigation in that direction. PMID- 11936355 TI - Immune system and surgical site infection. AB - It is well known that altered host defenses in general can play a significant role in the development of infection in any patient but their role becomes even more important in surgical patients where infections are the result of individual risk factors associated with the patient and other specific factors associated with the surgery itself. Thus all possible factors that can help modify the immune response should be taken into account in order to intervene, whenever possible, with therapies based on defined abnormalities to reduce the rate of post-surgical infections. Many factors associated with the patient have been clearly identified as responsible for a decreased immune response: old age, concomitant diseases (diabetes, renal and liver failure, solid and hematologic neoplasias, malnutrition, autoimmune diseases, AIDS) and concomitant therapies (corticosteroid, cytotoxic agents). Old age can affect both humoral and cell mediated immune responses. Chronic diseases can be responsible for a reduced primary response or depression of delayed hypersensitivity reactions (renal failure, neoplasias) or changes in leukocyte function (diabetes, leukemia, lymphomas). Malnutrition frequently accompanies diseases such as cancer, chronic and acute pancreatitis, inflammatory bowel diseases. Deficiencies in important vitamins and minerals (B6, A, folate, biotin, riboflavin...) can alter significantly the leukocyte function and immune response. Finally, there appears to be innate immune-suppression following any form of injury which is correlated with its magnitude and can affect any aspect of immunity. This has been well studied both in burn and surgical trauma. Alteration of phagocytosis, opsonization and chemotaxis are typically affected in burns, whereas surgical stress can include some reduction of cell mediated immunity. The best approach today to minimizing post-surgical infections is probably, besides use of antibiotic prophylaxis, to reduce the surgical trauma which consequently reduces the stress response and immune-suppression and to optimize the immune response by maintaining homeostasis through nutritional support. PMID- 11936356 TI - Antibiotic prophylaxis in clean surgery: vascular surgery. AB - The commonest and most dangerous infective complication in vascular surgery is prosthetic graft infection. Despite the use of routine systemic antibiotic prophylaxis, graft infection occurs after 3-5% of all prosthetic vascular reconstructions. Infection is associated with a high rate of major morbidity and mortality, with significant time and cost implications. Management can be difficult and the outcome is often disappointing. Data from Italian Registry of Prosthetic Graft Infections show that the commonest site of infection is the aorto-femoral district and that involved bacteria are usually gram-positive in early and low-grade infections and gram-negative in late and high-grade infections. Results are poor, with a mortality rate of 15% for elective surgery and of 60% for emergency interventions. We report the results of a multicenter randomized controlled trial of rifampicin-bonded Dacron grafts in aorto-femoral surgery, in which our Department was involved. Data demonstrate a reduction in total early wound and graft infection rates, and 2-year results show a small, non significant reduction in graft infection (1.7% in study group, 2.3% in control group). The same results were obtained in two other multicenter trials. PMID- 11936357 TI - Infection and chemotherapy: recent experiences, proposed strategies and perspectives in plastic and reconstructive surgery. AB - In plastic surgery the necessity of avoiding as much septic complication as possible is caused by major technical peculiarities. On the one hand the use of autologous grafts (skin, cartilage, bone, microsurgical flaps, etc.) makes the surgical act prone to infection, even in the case of "clean surgery" with almost total failure. On the other hand, reconstruction is performed in "unclean" and immunodepressed patients such as burned and polytraumatized patients. Finally, there are recent and extensive uses of normal and "smart" and biodegradable biomaterials (from silicon to hyaluronic acid esters, etc.) and particularly the recent possibility of applying the products of tissue engineering (cultured fibroblasts, cheratinocyte, chondrocytes grown and expanded in vitro on various scaffolds) with the known increase in infectious risks, has made mandatory the prevention and treatment of infections and also the identification of the "local" environmental increase in resistant strains. A brief review is presented together with the results of the Italian Multicenter Burns Antibiotic Protocol on more than 300 patients and the data on 284 patients treated with auto-semi-artificial skin expanded in vitro at the Institute of Plastic Surgery of the University of Milan Medical School. PMID- 11936359 TI - Introduction: Strategies for prevention and treatment of sepsis in post-surgical intensive care. PMID- 11936358 TI - Guidelines on antimicrobial chemotherapy for prevention and treatment of infections in the intensive care unit. AB - Severe infections (SIs) in Intensive Care Units (ICUs) constitute difficult therapeutic problems confronting clinicians who deal with severely ill patients. Some SIs are opportunistic infections acquired either in the community or in hospitals, particularly in immunodepressed patients. The great majority of ICU infections are of nosocomial origin. Resistant organisms have led to changing antibiotic therapy in ICU infections. Before microbiology is available, empiric therapy is based on: (i) proper identification of bacterial risks in each infection site; (ii) local surveillance of frequent nosocomial organisms/susceptibility patterns in the ICU; (iii) identification of environmental risk factors and the patient's underlying condition. In documented infection, antibiotic therapy must take into account gram-positive vs gram negative bacteria or mixed infections, pharmacokinetics/pharmacodynamic parameters of chosen antibiotic(s) and concentrations at the infection site, in order to prevent selection of resistant mutants and to provide the most efficient antibiotic therapy. With increasingly sophisticated intensive care measures, invasive exploratory procedures, and surgical procedures, evolving profiles of hospital infections require updated Guidelines for treatment of severe infections in ICUs. Preventive and therapeutic strategies include control of antibiotic use, and suitable antibacterial treatments which result in shortened hospital stay, improved outcome of hospital infections and significant cost savings. PMID- 11936360 TI - Mechanisms of tissue injury in sepsis: contrasts between gram positive and gram negative infection. AB - Sepsis is the systemic response to infection, and is caused by gram positive and gram negative bacteria in approximately equal numbers. In recent years it has become increasingly clear that there are significant differences between sepsis caused by gram positive and gram negative infections, both in the microbial components that initiate the injury, and the host responses that follow it. These differences will assume greater importance in developing targeted therapeutic interventions to severe sepsis. PMID- 11936361 TI - Antibiotic induced endotoxin release and clinical sepsis: a review. AB - Sepsis and peritonitis have not lost much of their danger for patients. The mortality rate in peritonitis has only marginally decreased during the last 30 years despite aggressive surgical and sophisticated intensive care treatment. In intra-abdominal infection and peritonitis source control remains the mainstay of treatment, although general principles and denominators of successful source control need to be established. Endotoxin has been recognized as a major player in the pathogenesis of sepsis and its significance in clinical disease has been investigated in clinical studies for more than 20 years. Since the Sixties there is a growing interest in the effect of antibiotics and other compounds on the release of endotoxin. The effect of antibiotics on the release of endotoxin and inflammatory parameters, e.g., cytokines, remains to be clarified despite a growing body of in-vitro studies, animal studies and a few clinical studies. The purpose of this review is to evaluate the evidence of endotoxin release in clinical studies and the effect that antibiotic treatment may have in-vitro, in vivo and in clinical studies on endotoxin and cytokine release. In-vitro antibiotic-induced endotoxin release may depend on antibiotic class, presence of serum, type of organism, site of antibiotic action and Gram-stain. Endotoxin release may be different in late or early lysis, proportional to the number of killed pathogens. Morphology of bacteria may have an impact on endotoxin release and phagocytosis. Antibiotic-treated animals may show higher endotoxin levels with a higher survival rate than untreated animals. Plasma endotoxin may increase despite decreasing bacteremia. There may be a similar killing rate by different antibiotics but a difference in endotoxin release. Intestinal endotoxin does not necessarily correlate to the level of gram-negative bacteria. However, the alteration of the gut content by pretreatment may be associated with reduced endotoxemia and increased survival. Antibiotic-induced endotoxin release may be different depending on the type of infection, the location of infection, the virulence of strains, Gram-stain, mode of application and dosage of antibiotic. Different antibiotics may induce the release of different forms of endotoxin which may be lethal for sensitized animals. The combination of antibiotics with inhibitors of endotoxin or the pro-inflammatory response may be responsible for increased survival by decrease of endotoxin release. The clinical significance of antibiotic-induced endotoxin release is documented only in a few clinical disorders, e.g., meningitis, urosepsis. The difference in endotoxin release by PBP 2-specific antibiotics, e.g., imipenem, and PBP 3-specific antibiotics, e.g., ceftazidime, may not be visible in each study. Patients with increased multi organ failure (MOF) scores may profit from treatment with antibiotics known to decrease endotoxin. In conclusion, the clinical significance of antibiotic induced endotoxin release remains to be clarified. Type of pathogen and its virulence may be more important than recently suggested. gram-positive pathogens were just recently recognized as an important factor for the development of the host response. In case of fever of unknown origin in intensive care patients either failure of treatment, e.g., failure of source control in intra-abdominal infection, or a side effect of antibiotic treatment, e.g., endotoxin release, should be considered as a cause of the fever. PMID- 11936362 TI - Laparoscopic surgery and surgical infection. AB - Laparoscopic surgery (LS) has improved our knowledge of some aspects of surgical physiopathology. Other advantages include a lower incidence of postoperative infections, as evidenced by a lower inflammatory response which is related to a better preserved immune response to infection. But, the differential aspects of LS may influence the intraperitoneal environment and, in case of infection, must be evaluated in two different situations: during clean and potentially contaminated surgery or in the presence of established infection. The most important differential factors of LS are the pneumoperitoneum and the use of CO2. The influence of both these on the evolution of an intraperitoneal infecton has been of interest in recent years. Our department developed an experimental study with mice to evaluate the local and systemic inflammatory response to perioperative intra-abdominal contamination with a known inoculum of Escherichia coli. The animals were distributed in four groups: control, laparotomy, laparoscopy with CO2 pneumoperitoneum, and laparoscopy with wall traction. Peritoneal liquid and blood cultures such as peritoneal and systemic cytokine levels were analyzed. The results showed a better tolerance to perioperative contamination in LS groups while the CO2 pneumoperitoneum had no influence. But, in the presence of peritonitis, an elevated CO2 pneumoperitoneum can be dangerous and the operative time is an important factor to be considered. The literature is reviewed on the relationship between LS and surgical infection. PMID- 11936363 TI - Pathogenesis of septic shock: implications for prevention and treatment. AB - The innate immune system is in the vanguard of host defenses against infection. Recognition of invasive microbial pathogens is mediated by pattern recognition receptors on the surface of immune cells that recognize pathogen-associated molecular motifs. Considerable progress has been made in recent years in understanding how bacterial products initiate sepsis. In gram-negative sepsis, the LPS-binding protein (LBP), CD14 and the recently identified Toll-like receptor 4 (TLR4) are key molecules for the recognition of endotoxin (lipopolysaccharide, LPS) by cells of the myelomonocytic lineage. In gram positive sepsis, components of the bacterial cell wall (peptidoglycan, PGN; lipoteichoic acids, LTA) have been shown to activate myeloid cells through an interaction with a receptor complex composed of CD14, TLR2 and perhaps also TLR6 (PGN) or CD14 and TLR4 (LTA). By contrast, gram-positive exotoxins act as superantigens and directly stimulate T lymphocytes by cross-linking the MHC class II of antigen presenting cells to specific chains of the T cell receptor. Immune cells activated by microbial pathogens release numerous effector molecules, which orchestrate the innate and adaptive host defenses. Furthermore, bacteria and microbial toxins directly activate the complement and coagulation systems, which play an important part in the host defensive response. Severe sepsis and septic shock can be viewed as clinical manifestations of a failing innate immune response that ultimately results in an overstimulation of the physiological host response. The pathogenesis of sepsis is far more complex that was initially anticipated. However, combined research efforts of basic scientists and clinical investigators continue to provide critical information for the identification of novel therapeutic targets. The exciting results obtained recently with treatment strategies designed to correct coagulation abnormalities occurring during sepsis are an example of how research may ultimately translate into improved patient care. PMID- 11936364 TI - Treatment of documented and suspected neutropenia-associated invasive fungal infections. AB - Factors such as the intensification of anti-tumor regimens have enhanced both the depth and length of neutropenia and endorsed severe deficiencies in other immune systems. As a result, the risk of fungal infections has increased substantially. Clinicians should be aware of the possibility to enable a timely diagnosis because many of the problems in the management of invasive fungal infections during neutropenia are as much the consequence of diagnostic short-comings as of lack of therapeutic options. About 7% of all febrile episodes during neutropenia can ultimately be attributed to fungi, Candida and Aspergillus species being the paramount pathogens. Although the data in favor of prophylactic use of antifungals are not convincing, prophylaxis is still recommended in an attempt to protect particularly high-risk patients. Fluconazole still appears a suitable agent in recipients of a bone marrow transplant. Given the paucity of data, reappraisal of the value of empirical antifungal therapy is warranted. Amphotericin B with or without 5-flucytosine is considered the standard therapy for acute candidiasis with fluconazole as an alternative. Amphotericin B is also first-line therapy for invasive aspergillosis in neutropenic patients; lipid based formulations are recommended for patients who develop nephrotoxity. Recovery of the granulocytes and other immune systems has shown to be of critical importance in the management of all invasive fungal infections. PMID- 11936365 TI - Antibiotic prophylaxis and treatment of surgical abdominal sepsis. AB - The incidence of surgical abdominal sepsis is related to the operation, patient and skill of the surgeon, ranging from <2-3% for laparoscopic cholecystectomy to >35-40% in overt peritonitis. Aged, obese, diabetic, neoplastic, acute patients have the highest incidence of sepsis. Antibiotic prophylaxis significantly reduces the incidence of postoperative infections for Class II and II operations. The proper timing (30-60 min before incision), choice of antibiotic (related to possible pathogens) and correct duration are essential. Ultra-short prophylaxis (only one administration) may be effective in most class II procedures and a cephalosporin can be used. Class II operations (colorectal) may require a booster dose soon after surgery or during surgery exceeding 3 h. The most effective regimen may include: ampicillin, clindamycin, I- II- III- or IV-generation cephalosporins, amoxycillin, aminoglycosides, metronidazole have been used alone and in combination. Combination prophylaxis should be active against aerobic and anaerobic bacteria. Treatment of surgical abdominal sepsis may be primary, seconday or tertiary. Surgery should remove the pathologic lesion, and antibiotics reduce the general effects of sepsis and infectious complications. This article presents information on the general rules for correct prophylaxis and treatment. PMID- 11936366 TI - Introduction: Nosocomial pneumonia in intensive care units. PMID- 11936367 TI - Epidemiology of major respiratory pathogens. AB - A vast literature attests to the fact that Streptococcus pneumoniae, Haemophilus influenzae and Moraxella catarrhalis represent the prevailing bacterial pathogens of community-acquired lower respiratory tract infections. Their specific incidence as causative agents of the more common syndromes is known to vary even profoundly, depending on geographic area, and the same holds true for their rates of resistance to antimicrobial drugs. Europe does not escape the threat posed by the present pandemic spread of penicillin resistance in S. pneumoniae although, as expected, some countries like Spain and France are highly affected and others including Germany, Italy, The Netherlands and the Scandinavian region, are relatively spared. In several sites multiple resistance has been described in S. pneumoniae with the most affected drugs being penicillin, the macrolides, co trimoxazole and tetracycline. In H. influenzae synthesis of beta-lactamases is the main resistance trait expressed. Lack of susceptibility to beta-lactams dictated by a different mechanism remains extremely rare. Large variations in the incidence of this character are apparent when considering European countries. France and Spain are again widely affected while Germany, The Netherlands and Italy display rates of beta-lactamase-positive H. influenzae of about 16%. M. catarrhalis must be considered generally resistant to non-protected aminopenicillins since over 90% of these organisms produce beta-lactamases. PMID- 11936368 TI - Ventilator associated pneumonia: risk factors and preventive measures. AB - Ventilator-associated pneumonia (VAP) is a common nosocomial infection associated with considerable morbidity and mortality. Various risk factors for VAP have been identified and include the duration of ICU stay and of mechanical ventilation, a diagnosis of trauma, and severity of illness. Knowledge of these factors can promote early diagnosis and hence treatment. In addition to simple, but very effective, basic hygiene, different preventative strategies have been suggested, and can be divided into those that aim to limit airway colonization, and those that improve host defense mechanisms. Of the former, non-invasive ventilation is effective but not always applicable or available, nursing the patient in the semi recumbent position is also associated with a reduced incidence of VAP but carries its own problems, stress ulcer prophylaxis remains controversial, and selective digestive decontamination is probably only relevant to certain subgroups of patients. Methods to improve host defense include early nutrition. Immunostimulatory therapies, such as interferon and granulocyte colony stimulating factor, require further research to confirm their place in the prevention or management of VAP. PMID- 11936369 TI - De-escalation antimicrobial chemotherapy in critically III patients: pros and cons. AB - In spite of advances in critical care, nosocomial infections still have a considerable impact on Intensive Care Unit (ICU) and hospital length of stay, mortality and costs. Several authors suggest that antibiotic therapy should be instituted as soon as sepsis is suspected in critically patients. Over the last two decades the rates of occurrence for pathogens have significantly changed under selective pressure from broad-spectrum antimicrobial therapy. Shifts from predominance of gram-negative to gram-positive organisms and outbreaks of resistant pathogens address the need for appropriate empirical regimens. Agents such as ceftazidime, imipenem and, more recently, meropenem and tazobactam have been used successfully as monotherapy. Two different clinical trials have reported that meropenem monotherapy is significantly more effective than ceftazidime-based therapy. Because of the outbreak of methicillin-resistant Staphylococcus aureus and Staphylococcus epidermidis, some investigators suggest adding a glycopeptide to beta-lactamase inhibitor and carbapenem as initial empirical therapy. Such a regimen should be administered before definitive proof of infections and until the results of microbial investigation are available (de escalation antimicrobial chemotherapy). On the other hand, several authors do not recommend glycopeptide administration in an attempt to limit nosocomial outbreaks of vancomycin-resistant enterococci (VRE) and staphylococci (VRS) and to avoid secondary drawbacks, such as nephrotoxicity and ototoxicity. De-escalation antimicrobial chemotherapy should be tailored to critically ill patients according to their clinical status, severity of illness and suspicion of sepsis or nosocomial pneumonia. PMID- 11936370 TI - Catheter-related infections: diagnosis and intravascular treatment. AB - The diagnosis of catheter-related infections relies on the presence of clinical manifestations of infection and the evidence of colonization of the catheter tip by bacteria, mycobacteria or fungi. The reference method to confirm the latter requires the withdrawal of the catheter for culturing, which frequently turns out to be inconvenient, unnecessary and costly. New methods try to avoid these inconveniences and to assess the presence of tip colonization without withdrawal. One of these methods uses quantitative blood cultures with a jump (> or = 5) in colony counts between blood obtained from the catheter lumen and simultaneously from a peripheral vein. It has a high sensitivity (>80%) and specificity (94% 100%) but is cumbersome and requires both an easy backflow of blood in the catheter and the existence of bacteremia. Cytocentrifugation and acridine orange staining of blood withdrawn from an infected catheter lumen has a sensitivity and a specificity of over 90% for the diagnosis of tip colonization. "Superficial cultures" consist in the semiquantitative culture of the hub, of the skin surrounding the catheter entrance and of the first (1 cm) subcutaneous portion of the catheter after swabbing. Sensitivity of this method is >90% and specificity is >80%, and positive and negative predictive values for catheters (considering together those with and without clinical data of infection) are 66% and 97%, respectively. Endoluminal brushing has turned out to be an impractical and unreliable procedure, at least in our experience. New methods based on the speed of bacterial growth to detectable levels of microorganisms in conventional blood cultures are a new and interesting way of assessing catheter-related infections. Besides, as use of antimicrobial-coated catheters becomes more prevalent, the existing definitions of catheter colonization and catheter-related infection may need to be modified, because such coatings may lead to false-negative culture results. Many catheter infections, diagnosed without catheter withdrawal, can be handled nowadays with the so-called "antibiotic lock-in technique", which consists in locking the infected catheter lumen with a solution containing antibiotics. A high proportion of infected catheters, mainly those with coagulase negative staphylococci, can be maintained in place and sterilized with this technique, including catheters in patients with therapeutic failure after receiving conventional intravenous antibiotic therapy. New diagnostic and therapeutic techniques may avoid the unnecessary withdrawal of thousands of efficient, difficult to replace and expensive intravascular lines. PMID- 11936371 TI - Cephalosporins in surgical prophylaxis. AB - Controlled clinical trials have shown that antimicrobial prophylaxis can lower the incidence of infection after certain operations, thus reducing morbidity, hospital stay, antibiotic usage and mortality due to sepsis. An effective prophylactic regimen should be directed against the most likely infecting organisms, but need not be active against every potential pathogen. Infection can be prevented when effective concentrations are present in the blood and the tissue during and shortly after the procedure. Therefore, antimicrobial prophylaxis should begin just before the operation: beginning earlier is unnecessary and potentially dangerous, beginning later is less effective. A single-dose prophylaxis after the induction of anesthesia is sufficient. If surgery is delayed or prolonged, a second dose is advisable if an antimicrobial drug with a short half-life is used. Postoperative administration is unnecessary and is harmful. Cephalosporins are considered to be the drug of choice, because they offer fewer allergic reactions. From the first generation cephalosporins, cefazolin has been widely recommended with success. From the second generation cephalosporins, cefuroxime, cefamandole and cefoxitin are increasingly recommended. Their antistaphylococcal activity is somewhat less strong but their activity against gram-negative bacteria is stronger. In addition, cefoxitin has good activity against anaerobes. Third generation cephalosporins, such as cefotaxime, cefoperazone, ceftriaxone, ceftazidime or ceftizoxime are generally not recommended for surgical prophylaxis. Despite these recommendations, they have been accepted by the medical community and are today in use in many countries as the most common drugs in surgical prophylaxis. Ceftriaxone in particular, is far exceeding the sales of any other drug for prophylaxis. Contra indications, limitations, additional or other drugs and practical recommendations for specific procedures are discussed and the results of several prospective randomized studies are presented. PMID- 11936372 TI - The prevention of central venous catheter-related sepsis. AB - Intravascular catheters continue to be a major source of sepsis which range from localised inflammation at the site of insertion to septicaemia. There are two main approaches which have been taken in an attempt to prevent these infections. These can be divided into the patient and the catheter. Approaches for prevention with patients include improved aseptic techniques, appropriate choice of antiseptic skin preparations, use of semi-occlusive dressings and cleaning schedules when using the devices. Catheters have also been developed to reduce the incidence of these infections. These include the production of polymers with smooth, anti-adhesive coatings, eg Hydromer, and more recently the introduction of devices which are either coated with antiseptics or antimicrobials. The antimicrobial coatings include antibiotics such as rifampicin and doxycycline or antiseptics, for example benzalkonium chloride or chlorhexidine. The efficacy of these approaches will be reviewed based on both laboratory and clinical studies. The treatment of intravascular catheter-related sepsis depends primarily on establishing the diagnosis. When the diagnosis has been made treatment ranges from the use of antimicrobials alone to removal of the device. More recent approaches have included the use of antibiotic locks within the catheters. Treatment schedules will also be reviewed. PMID- 11936373 TI - Novel strategies to prevent catheter-associated infections in oncology patients. AB - Aggressive cytotoxic treatment of cancer contributes to the growing number of life-threatening infections. Vascular catheters create predominant risks for staphylococcal, enterococcal and candida blood stream infections. Although the contaminating microorganisms may be few in number, the altered host immune response in the presence of such implants as well as disease-associated immunosuppression implies that even small bacterial counts have to be regarded as highly virulent species. Diagnosis of catheter-related infection (CRI) remains difficult before withdrawal of the suspected catheter. Positive culture of catheter surface, lumen and hub and positive peripheral blood probes (paired quantitative blood culture) are predictive for catheter related bacteremia (CRB). Diligent catheter care and effective antimicrobial catheters may reduce prolonged hospital stay, increased morbidity or mortality and serious economical consequences. The most promising approach features the incorporation of antimicrobial drugs into the polymer matrices that entrap but do not bind the drugs, allowing for extended release. For the efficacious prevention of colonization in the microenvironment of the implantable device the concentration of the antimicrobial substances must exceed usual antibiotic concentrations by a thousand-fold. This is the desired effect--high concentration near the device surface and very low systemic concentration. Incorporation of antimicrobials in the bulk material that constitutes a device can be effective as shown in several in vitro and in vivo studies. In the future, modification of both short-term and long-term catheters by biofilm-active antimicrobials creating slow delivery systems may provide an effective method to protect patients from nosocomial infection in oncology. PMID- 11936375 TI - Antimicrobial prophylaxis for abdominal surgery: is there a need for metronidazole? AB - The objective of the present multicenter observational study was to assess postoperative infections as a function of risk factors and antibiotic prophylaxis under everyday clinical conditions. 2513 patients from 114 centers in Germany who received infection prophylaxis prior to elective colonic resection were included in the study between 1st September 1996 and 30th September 1997. In the descriptive analysis of the study it was noted that 36.1% of the patients had received no prophylaxis with metronidazole despite the fact that the study protocol recommended the use of this drug in preoperative antibiotic combinations. The present analysis therefore considers the influence of metronidazole on the postoperative infection rate. To exclude any bias due to intergroup differences in risk profile, the groups with and without metronidazole were subjected to a matched-pair analysis. Matching parameters were: duration of operation, blood loss, age, diabetes mellitus, hepatic, renal, or chronic airways disease, immunosuppressive therapy, and rectal resection. This led to the formation of 800 pairs that were matched with respect to these parameters. The 800 pairs were then stratified into the following treatment groups: Group 1: long acting cephalosporin (ceftriaxone) with or without metronidazole (n = 2 x 491); Group 2: short-acting cephalosporins with or without metronidazole (n = 2 x 133); Group 3: broad-spectrum penicillins with or without metronidazole (n = 2 x 176). In all three treatment groups combination therapy with metronidazole was found to be significantly superior. Postoperative infection rates with and without metronidazole were 9.4% and 18.7% respectively in Group 1, 12.0% and 25.6% respectively in Group 2, and 19.9% and 29.0% respectively in Group 3. The lowest infection rate was thus achieved by means of preoperative infection prophylaxis with ceftriaxone plus metronidazole. Thus, preoperative administration of metronidazole in addition to a long-acting beta-lactam antibiotic is strongly advised in elective colon surgery, as absence of antibiotic cover against anaerobic colonic flora leads to a significantly higher postoperative infection rate. PMID- 11936374 TI - A multicenter study on central venous catheter-associated infections in Italy. AB - In a 1-year multicenter study the microbial colonization of 1154 central venous catheters (CVCs) was investigated. Catheters explanted either from immunocompromised or immunocompetent patients were collected and analyzed by five clinical microbiology laboratories located in Ancona, Aviano, Catania, Pavia and Rome, Italy. A further aim was to investigate, by scanning electron microscopy, the features of currently used catheters, both new and explanted from patients, analyzing their surface quality, the influence of the host protein biofilm on their microbial colonization, the modifications caused by their permanence in the body and the relationship between these factors and the occurrence of infections. PMID- 11936376 TI - A brief overview of the 1999 CDC Guideline for the Prevention of Surgical Site Infection. Centers for Disease Control and Prevention. AB - This summarizes many of the topics covered in the 1999 CDC Guidelines for the Prevention of Surgical Site Infection. It does not attempt to cover everything in the guideline. This article discusses many of the topics and lists most of the guideline recommendations. The guideline is a very in-depth, detailed publication with many references, tables, and graphs. All references/issues discusses in this overview are in the guideline, and the guideline is the only reference for this paper. PMID- 11936377 TI - Introduction: Guidelines for prevention and treatment of infection in orthopedic prosthetic surgery. PMID- 11936378 TI - Risk assessment for surgical site infections following total hip and total knee prostheses. AB - The purpose of the study was to assess the relative importance of risk factors for surgical site infections (SSI) following total hip and total knee prostheses in The Netherlands. In the period 1996-99 63 hospitals in The Netherlands registered SSI after 36,629 orthopedic operations. Total hip and total knee prostheses were analyzed in detail. The results of our study showed that a long preoperative stay was a risk factor for deep SSIs after both procedures. A dirty or contaminated wound and a serious systemic condition were risk factors for deep SSIs after total hip prostheses. If post-discharge surveillance was carried out, more SSIs were found, for total knee prostheses more than twice as many. Independent risk factors for SSIs after total hip prostheses were a contaminated/dirty wound and for total knee prostheses a short operation duration. The authors conclude that surveillance of surgical site infections following total hip and total knee prostheses revealed different risk factors for (deep) SSIs. PMID- 11936379 TI - Antibiotic prophylaxis in orthopedic prosthetic surgery. AB - Staphylococci account for approximately 70% of postoperative infections in orthopedic prosthetic surgery, with the leading organism being Staphylococcus epidermidis. Therefore, the antibiotics most widely used for prophylaxis are cefazolin, cefamandole and cefuroxime, by virtue of their excellent activity against these pathogens. However, methicillin-resistant coagulase-positive and negative staphylococci are increasingly being reported as the causative agents of postoperative infection in clean prosthetic surgery, therefore prompting the use of glycopeptide antibiotics (vancomycin and teicoplanin) in the prophylaxis for total joint replacement, particularly in hospitals in which there is high methicillin-resistance among these pathogens. Furthermore, in total primary knee replacement (TKR) performed with tourniquet application, regional prophylaxis with teicoplanin (i.e. the injection of teicoplanin into a foot vein of the leg to be operated on after occlusion of the systemic circulation by inflating the tourniquet) has been shown to be a safe and valuable prophylactic technique. Apart from providing teicoplanin concentrations in the operative field higher than those achievable by conventional systemic prophylaxis (i.e. by injecting the antibiotic into an arm vein before application of the tourniquet), regional prophylaxis was found in a prospective, controlled, open study on patients undergoing primary monolateral or bilateral TKR to result in a rate of postoperative infection similar to those achievable with conventional prophylactic regimens. PMID- 11936380 TI - Introduction: Parameters for prevention of post-operative infections. PMID- 11936381 TI - Two-stage exchange hip arthroplasty for deep infection. AB - Although the risk of infection after total hip arthroplasty has decreased over the last three decades with the use of prophylactic antibiotics, laminar airflow operating rooms and whole-body exhaust suites, deep infection after total hip arthroplasty remains a serious complication. Significant morbidity to the patient and the cost to the health care system remain. During this period of time, diagnostic techniques also have improved including the use of polymerase chain reaction amplification. Treatment options now include: suppressive antibiotics, irrigation and debridement with retention of components, one-stage reimplantation, two-stage reimplantation, and salvage procedures. Based on the medical literature, the successful eradication of a total joint replacement infection with a two-stage reimplantation protocol is over 90% while the success rate with a one-stage protocol is approximately 80%. These success rates may decline however as the prevalence of antibiotic resistant organisms increases. Current controversies regarding two-stage reimplantation protocols include: duration of intravenous antibiotic therapy, timing of the reimplantation, the use of allograft bone in the reconstruction, the choice of fixation (cement versus cementless), and the use of antibiotic-loaded cement spacers. PMID- 11936382 TI - Risk factors in surgery. AB - Improved surgical and anesthetic techniques and postoperative care have not significantly changed wound infection rates over the last 30 years. Many risk factors, related both to the host and to the surgical practice, have been identified in different studies. Control of nosocomial infections has become more challenging recently, due to a widespread bacterial resistance to antibiotics and to more frequent surgical indications in elderly patients at increased risk. A change in the microbiology of postoperative infections has also been noticed, characterized by a greater incidence of infections caused by methicillin resistant Staphylococcus aureus, by polymicrobic flora and by fungi. This paper reviews the most important risk factors encountered in general surgery, that we observed during a 6-year prospective study of wound infection carried out in our Department of Surgery at the University of Insubria in Varese. Furthermore, the epidemiologic data on wound infections recorded in 4,002 patients undergoing general surgical procedures (mostly gastrointestinal operations), are presented and discussed. PMID- 11936383 TI - Gentamicin PMMA beads and other local antibiotic carriers in two-stage revision of total knee infection: a review. AB - Local antibiotic carriers are available as resorbable carriers (e.g. gentamicin loaded collagen) and non resorbable carriers, mostly gentamicin-loaded bone cement in the form of beads or spacers. The release of the antibiotic may differ largely, depending on the properties of the carrier. In the use of spacers the release may remain too low, resulting in subinhibitory gentamicin concentrations. Revision of late deep infection of a knee prosthesis includes extraction, debridement and the use of local and systemic antibiotics. Reimplantation can be performed if healing is appropriate and the bone stock reaches. Otherwise arthrodesis is necessary. PMID- 11936384 TI - Preliminary results of a survey of the use of antimicrobial agents as prophylaxis in orthopedic surgery. AB - An epidemiological survey of the use of antimicrobial prophylaxis in Italian hospitals was carried out under the auspices of the Journal of Chemotherapy. Out of 500 Italian orthopedic centers queried, 225 agreed to participate in this study. A total of 136,321 surgical procedures were reported in the 166 centers reporting complete answers on type of surgery. They comprised hip and knee prosthesis (13.9%), spine surgery (4%), hip endoprosthesis (5.2%), osteosynthesis (26.9%), arthroscopy (24.4%), and others (25.5%). Perioperative antimicrobial prophylaxis was used in 75% of operations (ranging from 57.1% to 99.4% in arthroscopy and joint prosthesis, respectively). Short term (<24 h) antimicrobial prophylaxis was performed in 38.4% of the 206 centers answering this question correctly. 61.1% of centers employed single agent prophylaxis and 70.8% of these prescriptions were betalactam antibiotics. Bacteriological analysis revealed gram positive isolates in 73.3% of cases. Methicillin resistance was present in 45% of 915 tested strains. Out of 4221 patients with high risk of infectious complications (joint prosthesis surgery) given antimicrobial prophylaxis in 46 centers, the percentage of surgical wound infections was overall 2.1%, while that of non-surgical wound infections was 3.6%. The total infection rate was decreased by about half in association with long-term (>24 h) as compared to short-term (<24 h) antibiotic treatment (3.7% vs 7.6%, respectively), and with use of antibiotic drug combinations vs single antibiotic drugs (3.9 vs 6.6%, respectively). The incidence of surgical-site infection was not decreased by extending the chemoprophylaxis for more than the first 24 h after surgery, while it was reduced from 2.5 to 1.4% by use of combination antibiotic therapy. PMID- 11936385 TI - Introduction: Antibiotic prophylaxis in clean surgery. PMID- 11936386 TI - Microbial epidemiology patterns of surgical infection pathogens. AB - Resistance, as assessed in vitro, has a number of serious consequences in clinical situations. Treatment failures are common when an inappropriate drug has been prescribed and this, in turn, may lead to hospitalization of patients who normally would have been treated on an outpatient basis, as well as to longer hospital stay for inpatients and to the use of alternative drugs, which may be more expensive and more toxic. These factors all contribute to increased health care costs, morbidity and mortality. Microbiological procedures may identify the causative pathogen and provide the appropriate susceptibility pattern to the physician, thus reducing the chances of therapeutic failures. However, for a number of reasons including cost--even in hospitals--not to mention general practice, infections are seldom diagnosed on an etiological basis. From what has been stated, the knowledge of bacterial epidemiology and resistance represents basic support for correct therapeutic decision-making. PMID- 11936387 TI - The role of methicillin-resistant Staphylococcus aureus in orthopaedic implant surgery. AB - The problems presented by methicillin-resistant Staphylococcus aureus (MRSA) must be accommodated in both prophylactic and treatment regimens for orthopaedic implant surgery. The rationale of pre-admission nasal swabbing in directing prophylaxis for orthopaedic patients is discussed. The potential advantage of nasal mupirocin for Staphylococcus aureus and MRSA carriers is described. Methicillin-resistant Staphylococcus epidermidis is commented upon as another hazard in orthopaedics. Criteria for choosing glycopeptides in the treatment of implant infections are discussed, and need to be defined in orthopaedic units. Treatment regimens are briefly described. PMID- 11936388 TI - Antibiotic prophylaxis in clean surgery: clean non-implant wounds. AB - Wound infection after clean surgery (the majority being hernia, varicose vein and breast surgery) is often greatly underestimated. If a trained and blinded observer is involved using close and prolonged surveillance to at least 30 days postoperatively with appropriate definitions or wound scores, an infection rate of up to 15% or more may be found. Equally controversial is the value of prophylactic antibiotics in preventing postoperative wound infection; there is no clear cut evidence of efficacy and some random controlled trials (RCTs) have shown no differences at all. There is a need for guidelines to be drawn up but further RCTs may be needed. An alternative to antibiotics is the systemic warming of patients or the local warming of the operative site prior to surgery. In day case surgery in particular, patients may have been deprived of fluids for 12h prior to surgery; they may become cold whilst waiting for surgery dressed in a theatre gown; and their apprehension may not be controlled with anxiolytics. The inflammatory response may be obtunded with an increased risk of superadded infection or poor healing in cutaneous tissue resulting in wound separation and fat necrosis. Only a third of sampled purulent discharges grow skin organisms such as Staphylococcus aureus or epidermidis. In a study of 421 patients the 138 randomly assigned to local warming (Warm Up, Augustine Medical) had a wound infection rate of 3.6% compared with another group of 139 randomly assigned to systemic warming (Bair Hugger, Augustine Medical) of 5.8%. A standard-treated third group of 139 had a wound infection rate of 13.7% (P<0.001). The warmed patients also had significantly lower wound scores based on 4 systems, had higher skin temperatures and capillary flow prior to surgery and were prescribed fewer postoperative antibiotics by their family doctors in the 6 postoperative weeks (15.9% vs 6.5%; P=0.002). Wound infections are more common than generally accepted if they are looked for by close surveillance. Antibiotics may be avoided by the use of warming with the lessening of the risks of allergy, resistance and emergence. It is uncertain whether antibiotic prophylaxis has a role in clean wound surgery. Breast surgery carries the highest risk of wound infection and this may risk delay in receiving planned adjuvant chemotherapy or radiotherapy. Perioperative warming of the operative site may be of greatest value in this group of patients. PMID- 11936389 TI - Biomedical technology revolution: opportunities and challenges for oral and maxillofacial surgeons. AB - During this 45-minute presentation, I have tried to describe my vision of the exciting future that awaits us. I have tried to impart my enthusiasm for the opportunities provided to us as surgeons by the advances in molecular biology and genetics, imaging, surgical technology and bioinformatics. Most of all, I hope I have transmitted my optimism for the future to our younger members. I think the following statement or observation by the great educator Margaret Mead accurately summarizes our current situation regarding the application of all this new knowledge that will become available to us as surgeons: 'We are now at the point where we must educate people (surgeons) in what nobody knew yesterday, and prepare in our schools (training programs) for what no one knows yet but what some people must know tomorrow.' PMID- 11936390 TI - Use of the 'shark-fin' osteotome in separation of the pterygomaxillary junction in Le Fort I osteotomy: a clinical and computerized tomography study. AB - Le Fort I osteotomy fails in many cases to completely separate the pterygomaxillary junction and often results in fractures of the pterygoid bone and the tuberosity, which subsequently can cause complications. The objectives of this study were to describe the specifically developed Laster 'shark-fin' osteotome and to compare its use to other methods of pterygomaxillary dysjunction. Pterygomaxillary dysjunction was performed in 10 adult patients requiring Le Fort I osteotomy. In one randomly chosen side of the maxilla, the Obwegeser osteotome was used, while the Laster 'shark-fin' osteotome was used on the opposite side. A postoperative computerized tomography of the separation at the pterygomaxillary junctions revealed that in all sites treated with the Laster 'shark-fin' osteotome, a complete or almost complete separation was obtained, whereas the use of the Obwegeser osteotome resulted in five sites with fractures of the maxillary tuberosity and three with high-level fractures of the pterygoid plates (P<0.001). Comparing these findings with the literature, we concluded that the Laster 'shark-fin' osteotome is preferable for separating the pterygomaxillary junction in Le Fort I osteotomy. PMID- 11936391 TI - Sublingual gland involvement in systemic Wegener's granulomatosis: a case report. AB - Wegener's granulomatosis is a systemic vasculitis characterized by necrosis, granulomatosis and inflammation. The usual targets are extra-oral, although salivary gland involvement has been recognized, usually confined to parotid and submandibular glands. A case report is presented of sublingual gland presentation, confirmed on biopsy, which proceeded to systemic involvement. Some evidence suggests that salivary gland involvement, if recognized, may lead to an early diagnosis and the potential for earlier therapeutic intervention. PMID- 11936392 TI - Cerebellar infarct: secondary to arteriography for the diagnosis of a buccal hemangioma. AB - Angiography is a reliable diagnostic method for most of the vascular lesions. The method however is associated with complications and it is the aim of this report to present a cerebellum infarct resulting from angiography for the diagnosis of a buccal hemangioma. PMID- 11936393 TI - Traumatic myositis ossificans (ossifying pseudotumor) of temporal muscle. AB - The purpose of this paper is to present the clinical, radiographic, and histological aspects of a case of traumatic myositis ossificans of the temporal muscle. This is a rare, benign ossifying pseudotumor of skeletal muscles. It had developed 3 weeks after trauma to the left temple in a 33-year-old man. Differential diagnostic aspects of traumatic myositis ossificans confirming its benign character are discussed. PMID- 11936394 TI - Chronic eosinophilic pneumonia associated with T-cell lymphoma. AB - A case of a woman presenting with a parotid swelling is described. She had chronic eosinophilic pneumonia (CEP) for approximately 2 years prior to presentation. After further investigation she had biopsy of her parotid, which proved to be T-cell lymphoma. This is a rare association, the diagnosis and the management of this condition is discussed. PMID- 11936395 TI - Midfacial morphology in children with unilateral cleft lip and palate treated by different surgical protocols. AB - The purpose of this study was to compare cranio-facial, particularly midfacial morphology, in two groups of children with complete unilateral cleft lip and palate (UCLP) treated at two different cleft centres (Hannover. Germany and Brussels, Belgium) following different surgical treatment protocols. A total of 62 children (40 males; 22 females) with non-syndromic UCLP were included in this study at approximately the age of 10. The Hannover group comprised 36 children, who had repair of the lip at a mean age of 5.83 +/- 1.16 months, followed by repair of the hard and soft palate at a mean age of 29.08 +/- 4.68 and 32.25 +/- 4.29 months. respectively. The Brussels group consisted of 26 children who underwent surgical treatment according to the Malek protocol: the soft palate was closed at a mean age of 3.04 +/- 0.20 months, followed by simultaneous repair of the lip and hard palate at a mean age of 6.15 +/- 0.68 months. Midfacial morphology was evaluated by means of cephalometric analysis according to Ross. The children in the Hannover UCLP group did not differ significantly from those in the Brussels group in the anteroposterior dimension of the midface. However, the maxillary plane was significantly more open in the Brussels group due to less posterior vertical maxillary development. PMID- 11936396 TI - Analysis of errors in medical rapid prototyping models. AB - Rapid prototyping (RP) is a relatively new technology that produces physical models by selectively solidifying UV-sensitive liquid resin using a laser beam. The technology has gained a great amount of attention, particularly in oral and maxillofacial surgery. An important issue in RP applications in this field is how to obtain RP models of the required accuracy. We investigated errors generated during the production of medical RP models, and identified the factors that caused dimensional errors in each production phase. The errors were mainly due to the volume-averaging effect, threshold value, and difficulty in the exact replication of landmark locations. We made 16 linear measurements on a dry skull, a replicated three-dimensional (3-D) visual (STL) model, and an RP model. The results showed that the absolute mean deviation between the original dry skull and the RP model over the 16 linear measurements was 0.62 +/- 0.35 mm (0.56 +/- 0.39%), which is smaller than values reported in previous studies. A major emphasis is placed on the dumb-bell effect. Classifying measurements as internal and external measurements, we observed that the effect of an inadequate threshold value differs with the type of measurement. PMID- 11936397 TI - Risk factors of nerve injury during mandibular sagittal split osteotomy. AB - There is little objective data about whether surgical technique or mandibular anatomy are a risk for inferior alveolar nerve (IAN) injury during bilateral sagittal split osteotomy (BSSO). Orthodromic sensory nerve action potentials (SNAPs) of the IAN were continuously recorded on both sides in 20 patients with mandibular retrognathia during BSSO operation. Changes in latency, amplitude, and sensory nerve conduction velocity (SNCV) at baseline and at different stages of the operation were analyzed. The SNAP latencies prolonged, the amplitudes diminished, and the SNCVs slowed down during BSSO (P = 0.0000 for all parameters). The most obvious changes occurred during surgical procedures on the medial side of the mandibular ramus. There was a clear tendency towards more disturbed IAN conduction with longer duration of these procedures (right side R = -0.529. P = 0.02; left side R = -0.605, P = 0.006). Exposure or manipulation of the IAN usually had no effect on nerve function, but the IAN conduction tended to be more disturbed in cases with nerve laceration. Low corpus height (R = 0.802, P = 0.001) and the location of the mandibular canal near the inferior border of the mandible (R = 0.52, P = 0.02) may increase the risk of IAN injury. There was no correlation between the age of the patients and the electrophysiological grade of nerve damage. PMID- 11936398 TI - External neurolysis of the lingual nerve. AB - The lingual nerve is sometimes injured during the surgical removal of an impacted mandibular third molar. The level of sensory recovery was studied in 10 patients who underwent external neurolysis of the lingual nerve. The mean time from third molar surgery to neurolysis was 13.5 months (range 9-24 months). Seven of the 10 patients showed significant improvement, three patients regaining normal sensation. Three patients showed no improvement. These results show that external neurolysis should be considered for patients with altered sensation in the distribution of the lingual nerve. PMID- 11936399 TI - Groningen temporomandibular joint prosthesis. Development and first clinical application. AB - Patients with a severely degenerated temporomandibular joint (TMJ) may benefit from an alloplastic TMJ replacement. The aim of the study was to develop a safe and properly functioning TMJ prosthesis. The design was based on imitation of anterior condylar translation by an inferiorly located centre of rotation, unrestricted mandibular movements by a double articulation, correct fit to the skull by a self-adjusting skull part consisting of two connected parts, and stable fixation by bone screws that are rigidly connected to the prosthesis parts. The prosthesis consists of a titanium skull part with ceramic inlay, a titanium mandibular part with a ceramic spherical head, and an intervening polyethylene disc. Titanium-alloy bone screws are used for fixation. All parts are available in a number of different shapes. In vitro laboratory and in vivo animal tests showed a low wear rate, the possibility of a close fit to the skull, a stable fixation, sufficient mechanical strength, appropriate choice of materials and proper functioning. Thereafter the step to first patient application was made. First patient application was carried out without adverse events. In conclusion, the presented TMJ prosthesis passed the pre-clinical tests and has progressed to clinical application. The fit to the skull, the expected lifetime of the device and the reliability of the implantation procedure require further evaluation in well-designed clinical trials. PMID- 11936400 TI - Emerging patterns of the cervical cutaneous nerves in Asians. AB - Numerous reports exist upon clinical viewpoints of the four cervical cutaneous nerves. Unfortunately, a detailed description of the cervical cutaneous nerves has not yet been published. For this reason, administering effective anaesthesia to a particular nerve branch is difficult. The aim of this study was to clarify the anatomical knowledge about the emerging patterns of the cervical cutaneous nerves in the superficial neck using 35 Korean cadavers (22 male, 13 female). Four cervical cutaneous nerve branches penetrating the fascia of the posterior border of the sternocleidomastoid muscle (SCM) were classified into seven types based on the locations of their nerve emergence. Among these, the separated type (L-G-T-S) was the most frequent (50%). followed by the L-G x T-S type (20.3%), in which the great auricular nerve (G) and the transverse cervical nerve (T) emerged at the same level on the posterior SCM border. PMID- 11936401 TI - Diabetic ketoacidosis secondary to dento-alveolar infection. AB - Diabetic ketoacidosis (DKA) is a medical emergency with a potentially fatal outcome if not recognized and treated appropriately. Infective processes are a common precipitant of DKA. We report two cases of dentoalveolar infections in patients with type I diabetes mellitus who presented with DKA. The management of such cases requires both specialist surgical and medical intervention. PMID- 11936402 TI - Slow and continuous application of human recombinant bone morphogenetic protein via biodegradable poly(lactide-co-glycolide) foamspheres. AB - Bone morphogenetic proteins (BMPs) are multifunctional cytokines that were originally identified as molecules that induce bone and cartilage formation in vivo. In order to increase the efficacy of this potent protein for application in medicine, a carrier system is needed to retain the BMP at the preferred site. Here we present and characterize a slow-release carrier system for pure human recombinant (rh)BMP. The large porous microspheres, called 'foamspheres', are biodegradable, because they consist of poly(lactide-co-glycolide) acids and release loaded rhBMP slowly and continuously. In vivo studies in rodents revealed that rhBMP-loaded foamspheres increased the thickness of the calvarial bone of rats by 222%. When the same amount of rhBMP was applied via a gelatine-based hydrogel, the increase in bone height was only 66%. Thus, the carrier system for rhBMP is an important factor for the efficacy of BMPs. PMID- 11936403 TI - Effect of recombinant human bone morphogenetic protein-2 on bone formation in alveolar ridge defects in dogs. AB - This study was designed to evaluate the effect of recombinant human bone morphogenetic protein-2 (rhBMP-2) combined with poly D, L lactic-co-glycolic acid (PLGA)/gelatin sponge complex (PGS) on the formation of bone in critically sized marginal defects of the mandible in dogs. Three months after extraction of the pre-molar teeth, rectangular bone defects (10 x 8 x 7 mm) were made in both sides of the mandible. A PGS block soaked in rhBMP-2 (400 microgram/ml) was implanted into one defect (BMP (+) group). As control, an untreated PGS block was implanted into the contralateral defect (BMP (-) group). 2, 4, 8, and 12 weeks after implantation, the defects were examined. In the BMP (+) group, newly formed bone was found in all defects from 4 weeks onward and was marked at 12 weeks. In contrast, the BMP (-) group showed no appreciable new bone formation, even at 12 weeks. Moreover, density of newly formed bone in the BMP (+) group was similar to that of the surrounding cortical bone at 12 weeks. These findings suggest that rhBMP-2/PGS is an effective bone substitute for reconstructive surgery of the dog mandible. PMID- 11936404 TI - Evaluation of Bioglass/dextran composite as a bone graft substitute. AB - Allogenic and alloplastic bone graft substitutes serve either as bioinert or bioactive osteoconductors. Bioglass is a bioactive osteoconductor and also shows osteoproductive effects due to its high level of bioactivity. However, the material lacks some cohesiveness when used in augmenting certain bony surfaces, i.e. large or pleomorphic defects. The addition of medium molecular weight dextran modifies the particulate to a putty consistency and improves the handling characteristics. The objective of this study is to evaluate the influence of dextran upon the bioactive properties of Bioglass. Standardized bony defects in the lateral femoral condyles in adult New Zealand white rabbits were filled with one of five material groups: (1) autogenous bone; (2) Bioglass particulate; (3) Bioglass particulate mixed with dextran to a putty-like consistency; (4) a mixture of Bioglass and autogenous bone; (5) a mixture of Bioglass putty with autogenous bone. Postoperative healing was observed after periods of 2 days, 1, 2, 3, 6 and 12 weeks. Results showed no evidence of toxicity in the dextran containing materials, and defects in all test groups showed 100% bony ingrowth within 6 weeks. The addition of medium molecular weight dextran did not appear to alter the bioactive properties of Bioglass and had no adverse influence upon the ingrowth of bone into the defect sites. PMID- 11936405 TI - Combination of microplate and miniplate for osteosynthesis of mandibular fractures: an experimental study. AB - Because of the limited space available in the mandible, especially in the mental foramen and apical region, miniature osteosynthesis material is desirable. Recently, metal deposition in the direct neighbourhood of osteosynthesis plates made of titanium or even in peripheral organs have been reported in the literature with increasing frequency. The size and amount of osteosynthesis material used should therefore be kept to a minimum. In an experimental study on 60 models, we examined load-carrying stability and in a second series, torsional strength of two-piece plastic models connected by either a single miniplate, two miniplates, or a titanium microplate plus a miniplate. In our test arrangement, the average loading capacity of the combination miniplate/miniplate was 470 N: that of the combination microplate/miniplate was only 267 N. The test group with the single miniplate had an average loading capacity of only 225 N. Masticatory loads on the plates exceeding 200 N occur only 3 months after osteosynthesis. At that time the fracture has largely consolidated. The torsional strength of the microplate/miniplate combination was similar to that of the miniplate/miniplate combination (1,000 Nmm resulting in a width of the gap measuring 0.8 mm and 0.5 mm, respectively). The single miniplate was considerably less stable (0.8 mm gap width as early as with 300 Nmm). According to Champy, rotational forces in the anterior region of the mandible amount to approximately 1,000 Nmm and need to be withstood by the osteosynthesis material. Our results suggest that treatment of fractures in the interforaminal region with a combination of microplate and miniplate will be stable enough for early mobilization. PMID- 11936406 TI - Effect of local hyperthermia on metastases in oral squamous cell carcinoma. AB - In this experimental study, hamsters with oral squamous cell carcinoma (O-1N), which has a high potential for lymph node metastasis, received treatment with local hyperthermia. The effect of hyperthermia on regional lymph node metastases was examined pathologically. O-1N was heated twice, each session consisting of radiofrequency capacitive heating (13.56 MHz) for 40 min at 43 degrees C. Cervical lymph nodes were excised 14, 17, 21, and 28 days after heating and were examined histologically. Hamsters in the sham and control groups were killed on the same days and specimens were examined in a same manner. The incidence of lymph node metastasis was significantly lower in the hyperthermia group (36.4%) than in the sham (68.5%) and control (65.0%) groups (both P=0.02). The patterns of lymph node metastasis in the sham and control groups were more advanced than that in the hyperthermia group. The incidence of lymph node metastasis was very low (7.7%) in hamsters with no evidence of tumour after hyperthermia. On multivariate analysis, hyperthermia correlated with inhibition of cervical lymph node metastasis (P=0.02). Our findings suggest that local hyperthermia inhibits lymph node metastasis when the primary tumour responds histologically to treatment. PMID- 11936407 TI - Maxillofacial trauma due to work-related accidents. AB - Even though numerous reports on maxillofacial trauma exist, only a few give detailed information about work-related maxillofacial injuries. The purpose of this study was to reveal the significance of maxillofacial injuries related to accidents occurring at work by evaluating a large number of patients with maxillofacial injuries over a 9-year period. Out of the 8704 trauma patients treated between 1991 and 1999 in the Department of Oral and Maxillofacial Surgery at the University of Innsbruck, Austria, 463 (5.4%) were injured at work. All charts were reviewed and analyzed according to age, gender, cause of accident, occupation, type of injury, location and frequency of fractures. The highest incidence of maxillofacial injury was found among construction workers (a total of 124 patients, 26.8%), followed by craftsmen (102 patients, 22.0%) and office employees (69 patients, 14.9%). The sex distribution showed an overall male-to female ratio of 11.8:1 and those in the age group most affected were between 20 and 29 years of age. The most frequent cause of injury was a blow in 48.4%, followed by falls and falls over obstacles, accounting for 27.9% and 7.1%, respectively. Of all trauma, 45.4% (210 persons) sustained 423 maxillofacial fractures, 31.7% (147 patients) suffered 232 dento-alveolar injuries, and 21.2% (98 people) showed 430 soft-tissue injuries. One-fifth (20.7%) of all patients displayed concomitant injuries with cerebral and cranial trauma being the most common. The probability of sustaining maxillofacial trauma at work is correlated to the nature of the occupation. Individuals (mostly men) using tools or machines at work are exposed to a much higher risk of work-related maxillofacial trauma. PMID- 11936408 TI - Adhesive force: the underlying cause of the disc anchorage to the fossa and/or eminence in the temporomandibular joint--a new concept. AB - The commonly held cause for using the closed lock technique is the prevention of disc sliding by using the non-reducible disc. The purpose of this study was to re evaluate the pathogenesis of sudden and persistent severely limited mouth opening associated with a total lack of disc sliding in view of the fact that it is promptly released by lavage of the upper compartment of the temporomandibular joint (TMJ). The proposed pathogenesis is based upon earlier clinical and laboratory findings of the first author coupled with pertinent information culled from the literature. It is suggested that sliding of the disc in the TMJ is enabled due to the presence of phospholipids protected by hyaluronic acid (HA) that constitute an efficient lubrication system. Joint overloading may be associated with uncontrolled production of reactive oxygen species (ROS) that causes degradation of the HA, followed by the exposure of the phospholipids to lysis by phospholipase A2 (PLA2). The denuded, smooth and elastic articular surfaces that possess high surface energy become strongly adherent when placed in direct physical contact with each other. The presence of an extremely thin film of fluid (sub-boundary lubrication) between the mating surfaces may cause even higher adhesion. These adhesive forces are probably responsible for the flexible disc anchorage to the fossa and/or eminence. They also explain the immediate release of the disc and rehabilitation of its sliding following arthrocentesis. Since it is uncommon for two opposing surfaces to be stripped bare and to become adherent, the likelihood of anchored disc phenomenon (ADP) occurring and recurring is very low. PMID- 11936409 TI - FMD: DEFRA sets out its interim contingency plan. PMID- 11936410 TI - Studies of embryo transfer from cattle clinically affected by bovine spongiform encephalopathy (BSE). AB - Semen from 13 bulls, eight with clinical bovine spongiform encephalopathy (BSE), was used to artificially inseminate (AI) 167 cows with clinical BSE, and their resultant embryos were collected non-surgically seven days after AI. The viable and non-viable embryos with intact zonae pellucidae were washed 10 times (as recommended by the International Embryo Transfer Society) then frozen. Later, 587 of the viable embryos were transferred singly into 347 recipient heifers imported from New Zealand, and 266 live offspring were born of which 54.1 per cent had a BSE-positive sire and a BSE-positive dam. The recipients were monitored for clinical signs of BSE for seven years after the transfer, and the offspring were monitored for seven years after birth. Twenty-seven of the recipients and 20 offspring died while being monitored but none showed signs of BSE. Their brains, and the brains of the recipients and offspring killed after seven years, were examined for BSE by histopathology, PrP immunohistochemistry, and by electron microscopy for scrapie-associated fibrils. They were all negative. In addition, 1020 non-viable embryos were sonicated and injected intracerebrally into susceptible mice (20 embryos per mouse) which were monitored for up to 700 days, after which their brains were examined for spongiform lesions. They were all negative. It is concluded that embryos are unlikely to carry BSE infectivity even if they have been collected at the end-stage of the disease, when the risk of maternal transmission is believed to be highest. PMID- 11936411 TI - Economic aspects of controlling scabies on an open fattening farm with ivermectin in feed. PMID- 11936412 TI - Spastic syndrome in a bluefaced Leicester ram. PMID- 11936413 TI - Natural infection by a Babesia microti-like piroplasm in a splenectomised dog. PMID- 11936415 TI - Animal Health Bill. PMID- 11936414 TI - Use of moxidectin tablets in the control of canine subcutaneous dirofilariosis. PMID- 11936416 TI - Copycat or designer original? PMID- 11936417 TI - Providencia alcalifaciens in diarrhoeic dogs and cats. PMID- 11936418 TI - Postanaesthetic cerebral necrosis in five horses. PMID- 11936419 TI - RCVS council elections. PMID- 11936420 TI - Salt intake and hypertension therapy. AB - Hypertension is a risk factor for cardiovascular and renal organ damage. Environmental conditions affect the development of high blood pressure (BP), although genetic influences are also important. Current international guidelines recommend reducing dietary sodium to no more than 100 mmol (about 2.4 g sodium or approximately 6 g salt) per day to prevent BP rising; the current intake of sodium in industrialized countries is approximately double the recommended amount. Clinical trials (DASH and TOHP studies) have shown that dietary factors are fundamental in the prevention and control of BP. Low dietary sodium intake is particularly effective in preventing hypertension in subjects with an increased risk such as the overweight, borderline hypertensives or the elderly. A low-salt diet combined with anti-hypertensive therapies facilitates BP reduction independent of race. The hypotensive effect of calcium channel blockers is less dependent on salt intake than other drugs, such as ACE inhibitors or diuretics. Reduced sodium intake associated with other dietary changes (such as weight loss, and increasing potassium, calcium and magnesium intake) are important instruments for the prevention and therapy of hypertension. PMID- 11936421 TI - The impact of delayed graft function on the long-term outcome of renal transplantation. AB - Recent studies provide conflicting conclusions regarding the impact of delayed graft function (DGF) on the long-term outcome of renal transplantation. Some centres report DGF as an independent risk factor for reduced long-term graft and patient survival, while others report no impact on long-term outcome. Further scrutiny of data from these studies reveals differences in the definition of DGF, definition of long-term outcome, and statistical methods that may partly explain the variability. The commonest definition of DGF is the need for dialysis in the first week post-transplant, but this may be less informative than definitions that consider DGF as a continuous variable such as time to achieving creatinine clearance > 10ml/min. Acute rejection (AR) occurs more commonly in patients with DGF and variability in the impact of DGF may also relate to strategies to detect and treat AR during DGF. Centres with a vigilant strategy are likely to note a lower impact of DGF because the associated long-term adverse impact of AR is minimised. Furthermore, many centres reduce the dose of calcineurin inhibiting drugs and/or use polyclonal antibody therapy during DGF but the long-term impact of this strategy is unclear. Newer agents such as humanised anti-IL2 monoclonal antibodies and rapamycin may have a role, but controlled studies are required to define the optimal immunosuppressive regimen for patients with DGF. In the meantime, measures to minimise ischaemic damage to the transplant kidney and intensive surveillance for AR with weekly renal biopsy in patients with DGF are recommended. PMID- 11936422 TI - A decade of continuous improvement in cadaveric organ donation: the Spanish model. AB - BACKGROUND: There is an ever-increasing demand for cadaveric solid organs for transplantation all over the world. Partial strategies in many countries have resulted in small or transient increases in organ donation or even no improvement at all. In the early nineties, Spain started an original integrated approach designed to improve cadaveric organ donation. METHODS: In 1989 an official agency, the National Transplant Organization (ONT), and a national network of specially trained, dedicated and strongly motivated hospital physicians in charge of the whole process of organ donation, was created. The network now covers 139 hospitals. RESULTS: Cadaveric organ donors grew from 550 in 1989 to 1334 in 1999, a 142% increase, Cadaveric kidney transplants from 1021 to 2005 (96% increase), and total solid organs transplanted from 1302 to 3330 in ten years (156%). The rates of cadaveric organ donation per million inhabitants (33.6), kidney and liver transplantation (50.6 and 24.2) are the highest in the world. CONCLUSIONS: Spain is the only example in the world of continuous improvement in cadaveric organ donation registered in a large country over a ten-year period. Organ shortage is not due to a lack of potential donors, but rather to a failure to turn many potential into actual donors. A proactive donor detection program performed by well trained transplant coordinators, the introduction of systematic death audits in hospitals and the combination of a positive social atmosphere with adequate economic reimbursement for the hospitals have accounted for this success. This model can be partially or totally translated to other countries if basic conditions are satisfied. PMID- 11936423 TI - Cardiovascular risk in hypertensive patients: results of the Pandora project. AB - BACKGROUND: The aim of the Pandora project is to collect epidemiological information, check diagnostic and therapeutic pathways, and assess outcomes in a large hypertensive population. This report presents the results on patients enrolled in the study between 1997-1999. METHODS: Twenty-one general practitioners working in the Ravenna Local Health Service took part in the study. They were supplied with IBM compatible PCs and were trained to enter the patient's data (age, gender, familiarity for cardiovascular diseases, smoking, hospitalisations for cardiovascular disorders, diabetes, blood pressure, total cholesterolemia, creatininemia, antihypertensive therapy) on So.Ge.Pa. software. Cardiovascular risk factors were assessed according to the WHO - ISH joint committee recommendations. RESULTS: 2,608 treated hypertensive patients were enrolled, 65% of whom showed inadequate blood pressure control. The prevalence of inadequate BP control was higher in patients on multiple-drug antihypertensive therapy compared with those on monotherapy (71.9% vs. 47.9%), in older than in younger patients (70.7% vs. 56.1%) and in patients with three cardiovascular risk factors, or diabetes, or affected target organs, compared to those with two or less risk factors (72.4% vs. 63.3%), (p < 0.001 for all). 63.6% of patients were at risk for age, 36.6% for family history of cardiovascular diseases and 31.7% for severe hypercholesterolemia. CONCLUSIONS: BP control was inadequate in a large percentage of patients, but it was particularly unsatisfactory in the elderly and in patients with high cardiovascular risk. A cluster of cardiovascular risk factors was found in both adequately and inadequately controlled hypertensive patients. PMID- 11936424 TI - Temocapril, a long-acting non-SH group angiotensin converting enzyme inhibitor, modulates glomerular injury in chronic puromycin aminonucleoside nephrosis. AB - The purpose of the present study was to determine whether chronic administration of temocapril, a long-acting non-SH group angiotensin converting enzyme (ACE) inhibitor, reduced proteinuria, inhibited glomerular hypertrophy and prevented glomerulosclerosis in chronic puromycin aminonucleoside (PAN) - induced nephrotic rats. Nephrosis was induced by injection of PAN (15mg/100g body weight) in male Sprague-Dawley (SD) rats. Four groups were used, i) the PAN group (14), ii) PAN/temocapril (13), iii) temocapril (14) and iv) untreated controls (15). Temocapril (8 mg/kg/day) was administered to the rats which were killed at weeks 4, 14 or 20. At each time point, systolic blood pressure (BP), urinary protein excretion and renal histopathological findings were evaluated, and morphometric image analysis was done. Systolic BP in the PAN group was significantly high at 4, 14 and 20 weeks, but was normal in the PAN/temocapril group. Urinary protein excretion in the PAN group increased significantly, peaking at 8 days, then decreased at 4 weeks, but rose again significantly at 14 and 20 weeks. Temocapril did not attenuate proteinuria at 8 days, but it did markedly lower it from weeks 4 to 20. The glomerulosclerosis index (GSI) was 6.21 % at 4 weeks and respectively 25.35 % and 30.49 % at 14 and 20 weeks in the PAN group. There was a significant correlation between urinary protein excretion and GSI (r = 0.808, p < 0.0001). The ratio of glomerular tuft area to the area of Bowman's capsules (GT/BC) in the PAN group was significantly increased, but it was significantly lower in the PAN/temocapril group. It appears that temocapril was effective in retarding renal progression and protected renal function in PAN neprotic rats. PMID- 11936425 TI - Paradoxical rise in blood pressure during ultrafiltration is caused by increased cardiac output. AB - BACKGROUND: In some haemodialysis patients, blood pressure increases during the dialysis session despite ultrafiltration (UF). METHODS: We investigated six such patients who were not responsive to hypotensive drugs. Their echocardiograms were obtained prior to and during the dialysis session. RESULTS: After the mean 2520 +/- 1698 (4.5 +/- 2.3% of BW) ml of fluid was removed, the cardiac systolic function parameters significantly improved and maximum mean arterial pressure rose (from 107 +/- 5 to 118 +/- 6 mmHg, p < 0.027). This increase in blood pressure was accompanied by an increase in cardiac index (from 3.8 +/- 0.6 to 4.8 +/- 1.1 L/min/m2, p < 0.027). With continuing UF, after a mean fluid removal of 4133 +/- 1622 (7.5 +/- 2.1% of BW) ml, normal blood pressure was achieved in all patients. Previously increased ejection fraction and fractional shortening decreased. End-diastolic volume significantly decreased from 98 +/- 34 to 78 +/- 35 ml/m2 indicating normovolemia. This decrease in blood pressure was accompanied by a return of cardiac index to normal values (from 4.8 +/- 1.1 to 3.1 +/- 0.8 L/min/m2, p < 0.027). There was a positive correlation between mean arterial pressure and cardiac index (r = 0.56, p = 0.017). CONCLUSION: We hypothesize that our patients had passed the top of the Frank-Starling curve and were on the descending limb at the initial examination. With UF, patients first shifted to the left and upward on the curve. With further UF, they came down the ascending limb of the curve. In conclusion, paradoxical blood pressure rise during UF is caused by increased cardiac output, mediated by volume overload and can be treated by intensified UF. PMID- 11936426 TI - Predicting cardiac events with Tl201 dipyridamole myocardial scintigraphy in renal transplant recipients. AB - BACKGROUND: Renal transplant recipients should be screened for coronary artery disease. The purpose of this study was to evaluate the usefulness of thallium-201 dipyridamole myocardial perfusion single-photon emission computed tomography (SPECT) to predict major cardiac events in these patients. METHODS AND RESULTS: Eighty-two consecutive patients (61 males) in hemodialysis (HD) treatment, awaiting renal transplantation, underwent dipyridamole Tl-201 SPECT for pretransplant risk stratification. SPECT semiquantitative analysis showed a normal perfusion pattern in 52 patients (group A) and fixed defects in 9 (group B). Reversible perfusion defects were found in 21 (group C). Cardiac death, myocadial infarction, and unstable angina were sought as cardiac events. During a follow-up lasting 28.3 +/- 21.6 months, three patients of group A died for a non cardiac cause. In group C, six patients (28.6%) had a cadiac event: one had a non q wave myocardial infarction after renal transplantation; five were admitted with unstable angina. No cardiac events were observed in groups A and B (p = 0.0001). SPECT semiquantitative analysis showed that a summed difference score > or = 3 significantly increased the risk of cardiac events (odds ratio 4.5, C.I. 2.5 8.1). CONCLUSION: Cardiac events were only observed in HD patients with Tl-201 SPECT dipyridamole reversible defects (group C). A normal pattern or a fixed defect in Tl-201 dipyridamole SPECT in HD patients identified a good long-term prognosis. PMID- 11936427 TI - Emotional reactions and practical problems of the caregivers of hemodialysed patients. AB - BACKGROUND: The international literature consistently shows that the psychosocial outcomes of the informal carers (caregivers) of chronically ill patients are influenced by factors such as personality traits and perceived social support, but few studies have investigated these variables in the caregivers of hemodialysed patients, and the reciprocal experience of chronicity. METHODS: Fifty hemodialysed patients and their principal caregivers were recruited. They were administered specific questionnaires to evaluate their emotional stability and anxious/depressive reactions, the perceived burden related to the patients' condition, the quality of their family relationships and knowledge of the disease, and the degree of satisfaction with their lives. The study design was correlational and comparative. The data were analysed using Student's t test and Pearson's correlation. RESULTS: The patients were significantly more anxious and depressed than their caregivers, and had a more negative perception of their family relationships; they also had significantly higher neuroticism scores. Although the caregivers showed good emotional stability and a relatively low level of perceived burden, they stated that their daily lives were not very interesting and involved few social contacts. Twenty-five percent of them declared that they had financial problems; twelve percent also said they had to face problems of disease-related stigma and embarrassment. CONCLUSIONS: The results suggest that emotional stability is an important psychological determinant of perceived distress among the caregivers of hemodialysed patients. Assessing this personality trait and the reciprocal experience of chronicity in patients and caregivers may help nephrology teams identify subjects at major psychological risk, and to select the appropriate psychological support. PMID- 11936428 TI - Effects of clinical and individual variables on quality of life in chronic renal failure patients. AB - BACKGROUND: The purpose of this study was to assess the effects of baseline characteristics of patients, chronic renal failure and its treatment on quality of life. METHODS: To investigate the health status we used a questionnaire, the SF-36, during individual interviews. The study involved i) a cross-sectional observational stage, lasting from 1 Sept. 1997 to 30 April 1999, on all the patients undergoing conservative treatment, hemodialysis, peritoneal dialysis and kidney transplantation in district of Ravenna, and ii) a longitudinal stage, when the subjects were administered two questionnaires at an interval of at least 16 months. The quality of life data collected during the cross-sectional stage were analyzed to establish any correlations between age, type of treatment of chronic renal failure and hemoglobin levels. Patients under dialysis were also examined for any interference caused by the center where dialysis was done. RESULTS: Multivariate analysis on the data collected during the cross-sectional stage showed that chronic renal failure treatment and age affected the quality of life scores for the following parameters: physical activity, bodily pain, general health and vitality (age was inversely related to the scores). Transplanted patients and those on conservative treatment enjoyed the best overall quality of life, followed by those on peritoneal dialysis and those on hemodialysis. The physical domain scores were strongly associated with age, hemoglobin and diabetes. Multivariate analysis of the longitudinal study indicated that the interval between the beginning and the end of the observation period, age and diabetes greatly influenced quality of life. CONCLUSIONS: The data confirmed that age and diabetes have a strong influence on the quality of life and that the long period of treatment, with the absence of any prospect of resolving the clinical situation, has a negative effect on the quality of life in uremic patients. PMID- 11936429 TI - Carnitine action on red blood cell osmotic resistance in hemodialysis patients. AB - Low red blood cell osmotic resistance (RBCOR) in dialysis patients aggravates anemia and raises EPO needs. We studied RBCOR in 30 stable patients (22 M, 8 F), dialyzed for more than one year, with no systemic illness, blood transfusions, recent infection or treatment by ACE inhibitors. Nineteen were on EPO. Cuprophane dialysis membranes were used in 21, synthetic in the remaining nine. RBCOR was low in 13 patients and these patients received L-carnitine, 20 mg/kg IV, post dialysis, for one year (A); 17 with normal RBCOR served as untreated controls (B). We investigated the relations between RBCOR and membrane material, time on HD (THD), weekly dialysis duration (WHDT), serum total carnitine (TC), acyl carnitine (AC), free carnitine (FC) and AC/FC in all patients, before and after carnitine supplementation. RBCOR changes under carnitine treatment were evaluated. Age, sex, primary renal disease, THD, EPO dose, hematology and biochemistry were similar in treated patients and controls. Patients in group A (dialysed with cuprophane) had lower RBCOR than group B, (dialysed with synthetic and cuprophane membranes) (0.473 +/- 0.02 vs. 0.420 +/- 0.02, P < 0.001). RBCOR remained stable under carnitine in group A, but became abnormally low in controls (especially in 10/17 patients). RBCOR was significantly higher than the pretreatment values in 5/13 group A patients (month 0-M0: 0.48 +/- 0.02, M12: 0.45, +/- 0.02, P < 0.05). Carnitine levels, similar in both groups before treatment, remained stable in group A, but dropped in group B (TC: 52.1 +/- 9.6/31.1 +/- 21.2, FC: 33.1 +/- 8.3/15.6 +/- 19.2 micromol/L, P < 0.002). Patients with shorter WHDT (< or = 12 vs. > 12 h) had higher FC levels (36 +/- 6.9 vs. 30 +/- 6 micromol/L, P < 0.03). Low RBCOR is frequent in stable dialyzed patients. It is related to dialysis membranes and is aggravated by time on hemodialysis. Serum carnitine levels depend on weekly dialysis time and on carnitine supplementation, that normalizes osmotic resistance in some dialysis patients. PMID- 11936430 TI - Lymphocyte costimulatory receptors in renal disease and transplantation. AB - Cell-to-cell signal exchange during antigen presentation deeply influences the profile and extent of the immune response. Together with the TCR/MHC-mediated signal, accessory signals are provided to the T cell by the antigen-presenting cell (APC), through specific receptor-ligand interactions that represent indispensable costimulation for T-cell activation and survival. The main costimulatory pathways are the B7 family members and the CD40-CD154 receptor ligand pair. B7-1 and B7-2 costimulate T-cells by binding to CD28. Their binding is prevented by the neoexpression of CTLA4, a CD28 homologue that can deliver a negative signal. Another CD28-like molecule, called ICOS (inducible costimulator), has been described and binds B7RP-1, a third member of the B7 family, but not B7-1 and B7-2. The CD40-CD154 interaction works as a two way costimulatory system by triggering activation signals to both T-cell and APCs. Its importance is highlighted by the discovery that mutations of the CD154 gene are responsible for a severe human immunodeficiency. Disruption of the natural costimulatory interaction was highly effective for prevention and treatment in several experimental models of autoimmune disease and transplant rejection. This review focuses on the most significant advances in understanding the physiopathological events involving costimulatory molecules, and their impact on renal diseases and transplantation. PMID- 11936431 TI - Preeclampsia and fetal triploidy: a rarely reported association in nephrologic literature. AB - We report a case of a healthy woman - whose previous pregnancy was uncomplicated with early onset of hypertension, proteinuria and edema, during her second pregnancy. Ultrasound examination at 19th week of amenor rhea showed a fetus with growth retardation, corresponding to 17 weeks' gestation, ascites, cardiomegaly with serious multiple congenital anomalies. Amniocentesis for fetal karyotyping revealed 69, XXX. Because of continued elevated blood pressure, increasing proteinuria and severe lethal fetal anomalies, interruption of pregnancy was suggested. It was subsequently carried out by surgery. The patient underwent renal biopsy 10 days post-partum: histology showed the presence of the characteristic pathologic renal changes of preeclampsia. A year later, she became pregnant by the same partner. The third pregnancy was uneventful. The combination of fetal triploidy and preeclampsia may suggest a causative relationship. Clinically, most cases manifest as severe early-onset preeclampsia and must be differentiated from essential hypertension and a chronic glomerulonephritis (GN), which becomes symptomatic during pregnancy. When a fetus has triploidy, the counseling should stress the high incidence of preeclampsia; particularly when fetal anomaly is not compatible with life, it is well known that delivery of the fetus is curative in this syndrome. This information is important in counseling patients who are hesitant to terminate the pregnancy purely for a fetal abnormality, even if lethal. PMID- 11936432 TI - Nephrotic syndrome after allogeneic peripheral blood stem cell transplantation. AB - Nephrotic syndrome has been rarely reported after hematopoietic stem cell transplantation. We report a patient who developed nephrotic syndrome after allogeneic peripheral blood stem cell transplantation for acute myelogenous leukemia. Renal biopsy was performed and immunofluorescence and light microscopy were compatible with minimal change disease. The patient was treated with cyclophosphamide and prednisolone. Complete remission was achieved after three months. Previous reported cases are discussed. PMID- 11936433 TI - Anti-GBM nephritis complicating diabetic nephropathy. AB - There are several reports of glomerulonephritis (GN) in diabetics or patients with diabetic glomerulosclerosis. Cases of rapidly progressive GN where crescentic histologic changes are superimposed on diabetic glomerulosclerosis are very unusual. We report the case of a patient with type I diabetes mellitus, who developed rapidly progressive renal insufficiency. Renal biopsy disclosed anti glomerular basement membrane nephritis superimposed on classical diabetic glomerulosclerosis. PMID- 11936435 TI - Tuberculous meningitis in a renal transplant recipient. AB - Tuberculous meningitis is a very rare, but serious extrapulmonary complication of mycobacterial infections in immunocompromised patients, such as organ transplant recipients. We describe here a 66-year-old Turkish woman without any history of tuberculosis, who received a renal allograft transplant in 1994. After a pilgrimage to an endemic area for tuberculosis, she presented with fever and headache in August 1998. Clinical examination revealed positive meningism and hyperreflexia. Lymphocytosis was noted in her cerebrospinal fluid (CSF) and Mycobacterium tuberculosis infection was detected by PCR within the CSF. Despite immediate triple antituberculosis therapy, the patient's clinical condition deteriorated rapidly, with the development of septic shock syndrome, and she died three weeks after admission due to cardiovascular and respiratory failure. Mycobacterial infections, including extrapulmonary manifestations, should thus be considered in all renal transplant recipients presenting with unexplained fever. Preventive therapy, i.e. isoniazid prophylaxis, may also be recommended for patients risking exposure in areas endemic for tuberculosis. PMID- 11936434 TI - IgM antibodies against cytomegalovirus in SLE nephritis: viral infection or aspecific autoantibody? AB - Despite many studies on the subject, the causal relationships between viruses and presentation/exacerbation of autoimmune diseases are still elusive. The possibility of false positive IgM antibody tests for human cytomegalovirus (CMV) in patients with systemic lupus erythematosus (SLE) has been pointed out. Here we report a case of a patient who developed lupus nephritis, with biochemical and clinical markers of CMV infection with intestinal involvement. At first, the antibodies to CMV were regarded as spurious aspecific signs of autoimmune disease. The patient had had serious flare-ups of the disease, hemolytic-uremic syndrome with thrombotic microangiopathy superimposed on SLE nephritis, and life threatening infections for three years until CMV infection was confirmed by the persistence of anti-CMV IgM-antibodies coupled with positive results of tests for viral replication. After therapy with ganciclovir, his clinical and biochemical condition improved and remained stable for three years, with only very low maintenance steroid coupled with hydroxychloroquine. IgM anti-CMV were no longer detectable in spite of the persistence of other autoantibodies such as anti-DNA and ANA. Keeping in mind that CMV-IgM has been reported in only 5% of patients with SLE nephritis, the history of our patient indicates that CMV infection must be carefully excluded before IgM antibodies against CMV can be simply classified as an aspecific sign of cross-reacting autoantibodies formed in SLE patients. PMID- 11936436 TI - Prevalence of astigmatism among students in northern Greece. AB - PURPOSE: To assess the prevalence of astigmatism in a sample of 1738 students (15 18 years old) from Northern Greece. METHODS: Collection of the sample was based on a questionnaire method. Statistical analysis included estimation of the prevalence of astigmatism and the distribution of students according to their cylindrical values. We also checked whether heredity or sex affected the occurrence of astigmatism. RESULTS: The prevalence of astigmatism was 10.2%. It was mostly at low levels, up to 2 D cyl. Females ran a significantly higher risk of astigmatism than males, and heredity seemed to be an important predisposing factor for this refractive error. CONCLUSIONS: There is considerable variability in the prevalence of astigmatism worldwide, as indicated by different studies. However, this refractive error prevails at low levels in the Greek student population, compared with other countries. PMID- 11936437 TI - Comparison of tear function tests and impression cytology with the ocular findings in acne rosacea. AB - PURPOSE: The aim of this study was to determine the correlation between tear function tests and impression cytology results and the clinical findings in ocular rosacea. METHODS: Patients with a histopathologic and clinical diagnosis of acne rosacea were studied. Ocular examination consisted of best-corrected visual acuity measurement, slitlamp examination, tear break-up time (BUT), basal Schirmer test, and rose Bengal staining. Impression cytology was done two days later. Ocular findings and symptoms were checked and scored. Age-matched controls were assessed using the same parameters. RESULTS: The most frequent symptoms were itching (80%), redness (68%), burning (64%), and photophobia (60%). The most frequent clinical findings were meibomitis (92%), telangiectasis (88%), blepharitis (84%), superficial punctate keratopathy (72%), and hyperemia (60%). Mean tear BUT was 9.6+/-3.1 (SD) seconds (range 5-17 seconds). Mean Schirmer test measured 11.4+/-1.6 mm (SD) (range 8-15 mm). Mean rose Bengal staining scored 1.8+/-0.1. Mean tear BUT, Schirmer test and rose Bengal staining scores were lower in the rosacea group than the control group (p<0.003, p=0.04, p< or =0.038, respectively). Staining was pathologic in 18 patients (36%) and the highest staining score was 4. No significant difference was found between the stages of the nasal and temporal conjunctival impression cytologies in each eye, and there was no relationship between impression cytology stages and rose Bengal scores (p>0.05). A significant relation was observed between the stages of impression cytology and the severity of meibomitis (Fisher's test, chi2=9.625, p=0.001). The Schirmer test gave lower results in patients with severe blepharitis (Mann Whitney U Test, U=180.5, p=0.034). CONCLUSIONS: Early diagnosis prevents serious ocular complications and chronic dry eye in rosacea. We suggest that in addition to tear function tests, rose Bengal staining and impression cytology can be successfully used in the early diagnosis of dry eye and in monitoring medical treatment in ocular rosacea. Meibomian glands play an important role in the pathogenesis of the ocular disease. PMID- 11936438 TI - Corneal thickness in trachomatous dry eye. AB - PURPOSE: To investigate the effect of severe desiccation on corneal thickness in scarring trachoma by comparing the thickness of normal and trachomatous dry eye corneas. METHODS: Ultrasonic pachymetry was used to measure the corneal thickness at nine points in the central and peripheral cornea (superior, superonasal, nasal, inferonasal, inferior, inferotemporal, temporal, superotemporal) in 45 eyes of 27 patients with trachomatous dry eye and 54 eyes of 31 normal subjects. RESULTS: The average thickness of the nine sites in the central and midperipheral cornea was significantly less in trachomatous dry eyes than normal eyes. The superior cornea was the thickest area in both groups, measuring 574.03+/-31.62 microm in trachomatous dry eyes and 611.33+/-34.99 microm in normal eyes (p<0.001). The centre of the cornea was the thinnest, measuring 510.43+/-32.12 microm in trachomatous dry eyes and 546.27+/-36.20 microm in normal eyes (p<0.001). CONCLUSIONS: The thickness of the central and midperipheral cornea was significantly reduced in patients with trachomatous dry eye. The chronic state of severe desiccation, tear film instability and increased immune activation in trachomatous dry eye may contribute to this thinning. PMID- 11936439 TI - Increased corneal thickness in active Behcet's disease. AB - PURPOSE: To measure central corneal thickness (CCT) in patients with Behcet's disease (BD), particularly in the active disease. MATERIALS AND METHODS: We measured CCT by ultrasound pachymetry in 64 patients with Behcet's disease and in 20 healthy controls. Forty-one of the patients with BD had ocular involvement and 23 none; 19 of the 41 with ocular involvement were in an active period and 22 had inactive disease. RESULTS: The mean CCT in the group with active ocular involvement was significantly higher (589+/-27 microm) than in the control group (553+/-21 microm) (p = 0.003), the group with inactive ocular involvement (560+/ 26 microm) (p < 0.001), and the group with no ocular involvement (558+/-25 microm) (p < 0.001). After appropriate treatment of patients with active ocular involvement, the mean CCT returned nearly to normal (563+/-20 microm) and the difference from controls was not significant. There was no significant difference for the mean CCT between controls and the patients with no ocular involvement or with inactive ocular involvement. CONCLUSIONS: Behcet's patients with active ocular involvement have a thicker CCT than patients with inactive ocular involvement. There was no difference in CCT between controls and patients with no ocular involvement. The CCT of patients with active ocular involvement returned to nearly normal after treatment. CCT must be taken into account when developing a managing and following approach for Behcet's patients with active ocular involvement. PMID- 11936440 TI - Long-term evaluation of endothelial cell loss after phacoemulsification. AB - PURPOSE: To check whether three months' follow-up is sufficient to determine endothelial cell loss after cataract surgery and whether this value depends on preoperative central endothelial cell density. METHODS: A two-year prospective assessment of central endothelial cell density was done in 60 eyes after phacoemulsification. In 30 eyes a 3.5 mm corneal incision closed with a single crossed suture was made, and the other 30 had a 3.5 mm scleral tunnel no-stitch incision. Central endothelial cell density was measured before surgery and 1, 3, 6 months and 2 years after. RESULTS: The mean rate of endothelial cell loss even two years after surgery was significantly higher than the physiological rate, amounting to 0.9% per year. There was no correlation between preoperative central endothelial cell density and postoperative cell loss. CONCLUSIONS: Even two years' follow-up is not sufficient to establish the total endothelial cell loss after cataract surgery. The amount of cell loss does not depend on the preoperative density. PMID- 11936441 TI - Factors influencing treatment results in pseudophakic endophthalmitis. AB - PURPOSE: To evaluate risk factors, therapeutic approaches and factors associated with the poor visual outcome in pseudophakic endophthalmitis. METHODS: Data related to 28 cases with the diagnosis of endophthalmitis after cataract surgery and IOL implantation were gathered retrospectively. RESULTS: Preceding surgery was extracapsular cataract extraction (ECCE) in 18, phacoemulsification in 8 and scleral fixated intraocular lens implantation in two cases. Posterior capsule rupture and diabetes mellitus were considered to contribute to the development of endophthalmitis because of their high incidences (50% and 25%) in the study group. Microbiological studies from aqueous and vitreous humour were done in 85% of the cases and 58% were positive. S. Epidermidis was the most common organism, accounting for 50% of the isolates. All cases were given topical and systemic antibiotics. Inflammation was controlled by addition of subconjunctival antibiotics to this regimen in two, intravitreal antibiotic injection in 14, pars plana vitrectomy, total capsular and lens extraction and intravitreal antibiotic injection in three, lens exchange, intracapsular and intravitreal antibiotic injection in three cases. Six (21%) cases eventually needed evisceration. Visual acuity of 20/40 or better was achieved in 25%, and 20/100 or better in 64%. CONCLUSIONS: Treatment delay (p=0.039), capsular rupture complicating cataract surgery, especially with extracapsular cataract extraction (p=0.015), and initial visual acuity worse than hand motion (p=0.003) were strong predictors of poor visual outcome. The risk of endophthalmitis was not different forplanned ECCCE (0.26%) andphacoemulsification (0.27%) but the prognosis was better with the latter. PMID- 11936442 TI - Group II phospholipase A2 content of tears in patients with senile cataract and primary open-angle glaucoma. AB - PURPOSE: To determine the group II phospholipase A2 (PLA2) content of tears in patients with senile cataract or primary open-angle glaucoma (POAG) and to compare it with the PLA2 content of tears in age-matched healthy controls. METHODS: The PLA2 concentration of tears was measured with time-resolved fluoroimmunoassay in 21 patients with senile cataract, 23 patients with POAG and in 40 healthy controls. RESULTS: The PLA2 content of tears was 38.3+/-30.1 microg/ml in patients with senile cataract, 32.1+/-22.3 microg/ml in patients with POAG, and 36.6+/-31.1 microg/ml in healthy controls. There were no significant differences between the patient and the control groups. CONCLUSIONS: We conclude that neither senile cataract nor POAG has any effect on the PLA2 content of tears. PMID- 11936443 TI - Decreased nitric oxide production in primary open-angle glaucoma. AB - PURPOSE: Raised intraocular pressure (IOP) is the major risk factor responsible for optic nerve damage in primary open-angle glaucoma (POAG). The trabecularmeshwork acts as a valve in aqueous outflow and relaxes with nitric oxide (NO) agonists. Since NO is synthesized by endothelium and smooth muscle elsewhere in the body, this study investigated the NO levels in the aqueous humor of patients with POAG compared with cataract patients. MATERIALS AND METHODS: Aqueous humor samples were taken by paracentesis from 16 consecutive patients with POAG (9 male and 7 female; mean age 69.0+/-3.4 yrs) and 14 age and sex matched controls with cataract (8 male and 6 female; mean age 66.7+/-4.1 yrs) during elective surgery. As an indicator for NO, aqueous total nitrite levels (end - product of NO) were measured by Greiss reaction. The Mann-Whitney U test was used for statistical analysis and P <0.05 was considered significant. RESULTS: The mean age and sex in two groups were comparable. The mean aqueous humor NO levels were significantly (P = 0.001) lower in patients with glaucoma (72.72+/-11.21 micromol/L) than in patients with cataract and no glaucoma (86.92+/-11.23 micromol/L). CONCLUSIONS: Decreased NO production in patients with POAG indicates that NO-producing cells may be lost as the disease progresses. The control of NO levels in the eye might be a therapeutic target in glaucoma. PMID- 11936444 TI - Glaucomatous damage patterns by short-wavelength automated perimetry (SWAP) in glaucoma suspects. AB - PURPOSE: To determine the glaucomatous visual field damage patterns by short wavelength automated perimetry (SWAP) in glaucoma suspects, and to compare the frequency of diffuse visual field losses and localized defects. METHODS: 157 eyes of 157 ocular hypertensive subjects who met the selection criteria (intraocular pressure greater than 21 mm Hg and normal standard visual fields) were studied. SWAP was done with a modified Humphrey Field Analyzer. Total (TD) and Pattern Deviation (PD) probability maps were calculated for SWAP. The frequency of abnormlities in the TD and PD were determined, analyzing the visual field loss components. RESULTS: The involvement of the test points was more frequent on the TD plots than on the PD plots for all levels of defects (p< 0.001). The glaucomatous defects also showed certain topographical distribution. CONCLUSIONS: A diffuse sensitivity component of visual field loss was found at all SWAP defect depths in glaucoma suspects. PMID- 11936445 TI - Causes of blindness among adult Jordanians: a hospital-based study. AB - PURPOSE: To assess the causes of blindness among adults aged 20 years and over who attended two big general ophthalmic clinics in Jordan. PATIENTS AND METHODS: This hospital-based study was undertaken at Jordan University Hospital in Amman (the capital of Jordan) and Princess Basma Teaching Hospital which is affiliated with Jordan University of Science and Technology in Irbid, the third largest city in the country. A total of 2732 patients seen consecutively over 26 months were examined and the ophthalmic findings were recorded. Using standard Snellen charts, subjects were placed in one of three categories according to their best corrected visual acuity: (1) unilateral blindness: less than 6/60 in the worst eye, 6/60 or more in the better eye; (2) moderate bilateral blindness: less than 6/60 in the worst eye, less than 6/60 to 3/60 or more the better eye; and (3) severe bilateral blindness: less than 3/60 in both eyes. RESULTS: Of the totalpatients seen, 373 were blind according to the selection criteria. Among 248 patients with unilateral blindness, diabetic retinopathy, cataract and trauma were the leading causes. Among the 81 patients with moderate bilateral blindness, diabetic retinopathy and cataract were the leading causes. Diabetic retinopathy and glaucoma were the leading causes in patients with severe bilateral blindness. CONCLUSIONS: These data could be used in planning blindness prevention and treatment programs while awaiting a national survey on the prevalence and causes of blindness in Jordan. PMID- 11936446 TI - Outcome of abducens nerve paralysis in patients with nasopharyngeal carcinoma. AB - PURPOSE: The objective of this study was to investigate the incidence, management and outcome of patients with nasopharyngeal carcinoma (NPC) who developed Vlth nerve palsy. METHODS: Between December 1993 and December 1999, we investigated retrospectively the charts of 166 patients with NPC. RESULTS: Twenty-eight of 166 patients (16.8%) had cranial nerve involvement at the time of the diagnosis of NPC. Sixteen (57.2%) were identified as having abducens nerve palsy. In 25% abducens nerve palsy was the presenting symptom. Three patients were able to compensate for their diplopia after prism correction or botulinum toxin-A injection, and six (50%) completely recovered from abducens nerve palsy after either radiotherapy or chemotherapy. CONCLUSIONS: Abducens is the most common cranial nerve involved in NPC, radiotherapy and or chemotherapy relieves the paralysis in half the patients. Prism correction or botulinum toxin-A injection are effective non-invasive procedures for patients with significant diplopia. PMID- 11936447 TI - Indolent corneal ulcers in a patient with congenital insensitivity to pain with anhidrosis: a case report and literature review. AB - PURPOSE: To report a case of bilateral corneal neurotrophic ulcer in patient with congenital insensitivity to pain with anhidrosis (CIPA) and review the literature. CASE REPORT: A 6 year-old boy presented with bilateral central corneal sterile ulcer, decreased corneal sensitivity, moderately altered corneal reflex and normal tearing response. History taken, systemic evaluation and medical chart review were undertaken. DISCUSSION: Fifty-two cases of CIPA have been reported worldwide. Fourteen cases had corneal involvement. The clinical picture of our patient is characteristic of CIPA. CONCLUSIONS: Congenital insensitivity to pain with anhidrosis may present as neurotrophic corneal ulcer. We report herewith, this vision threatening corneal congenital abnormality. Early diagnosis and prompt treatment are mandatory to prevent corneal complications such as scarring and perforation. PMID- 11936448 TI - Non-penetrating deep sclerectomy in unilateral open-angle glaucoma secondary to idiopathic dilated episcleral veins. AB - PURPOSE: To investigate the efficacy of non-penetrating deep sclerectomy (NPDS) in a secondary open-angle glaucoma case due to dilated episcleral veins, on intraocular pressure and retrobulbar hemodynamics. METHODS: NPDS was done on a 32 year-old male unilateral open-angle glaucoma secondary to idiopathic dilated episcleral veins. RESULTS: Except for hyphema and shallow anterior chamber on the first post-operative day, no other complications were observed. Intraocular pressure was 18-20 mmHg during the first six months, without medication or visual loss. Retrobulbar color Doppler imaging showed normal mean and end-diastolic velocities and pulsatility indices of the posterior ciliary and central retinal arteries at end of the first post-operative year. CONCLUSIONS: NPDS may be an alternative to trabeculectomy in open-angle glaucoma secondary to dilated episcleral veins. PMID- 11936449 TI - Vitreous hemorrhage following phakic anterior chamber intraocular lens implantation in severe myopia. AB - PURPOSE: To describe two cases of vitreous hemorrhage following phakic anterior chamber lens (AC-IOL) implantation in high myopia. CASE REPORT: In case 1, hemorrhage developed one month after surgery, without retinal involvement, and visual acuity (VA) resulted 20/200 after pars-plana vitrectomy (PPV). In Case 2, vitreous hemorrhage was complicated by retinal detachment (RD). PPV and silicone oil injection were performed, with AC-IOL removal and cristalline lens extraction. After 2 years the retina was attached and VA was 20/80. DISCUSSION: Only few cases of RD, posterior uveitis and endophthalmitis are reported following phakic AC-IOL implant. Vitreous hemorrhage could represent an additional posterior segment complication. Intraoperative manoeuvres, hypotony induced posterior vitreous detachment and/or peripheral retina traction could play a role in engendering this complication in highly myopic eyes. PMID- 11936450 TI - Disorders of the temporomandibular joint. What comes next? PMID- 11936451 TI - Saliva--the defender of the oral cavity. PMID- 11936452 TI - Clinical applications of antimicrobial host proteins lactoperoxidase, lysozyme and lactoferrin in xerostomia: efficacy and safety. AB - Innate human salivary defence proteins, lysozyme, lactoferrin and peroxidase, are known to exert a wide antimicrobial activity against a number of bacterial, viral and fungal pathogens in vitro. Therefore, these proteins, alone or in combinations, have been incorporated as preservatives in foods and pharmaceuticals as well as in oral health care products to restore salivas' own antimicrobial capacity in patients with dry mouth. These antimicrobials used in oral health care products, such as dentifrices, mouth-rinses, moisturizing gels and chewing gums, have been purified from bovine colostrum. In this review I critically evaluate the clinical efficacy and safety of this kind of preventive approach against various oral diseases and symptoms. PMID- 11936453 TI - Understanding salivary fluid and protein secretion. PMID- 11936454 TI - Expression of cytokeratins (CKs) 8, 13 and 18 and their mRNA in epithelial linings of radicular cysts: implication for the same CK profiles as nasal columnar epithelium in squamous epithelial lining. AB - OBJECTIVE: Our objective was to specify cytokeratin (CK) patterns in lining epithelia of radicular cysts which are sometime lined with ciliated columnar epithelia as seen in the nasal epithelia. MATERIALS AND METHODS: We examined the CK expression in 52 radicular cysts obtained from 32 maxillary and 20 mandibular lesions and investigated CK-mRNA expression using in situ hybridization in 24 maxillary and 13 mandibular cysts and reverse transcription-polymerase chain reaction (RT-PCR) in 24 maxillary cysts. RESULTS: Of the maxillary cysts, 20, 29 and 19 squamous epithelial linings were positive for CK8, CK13 and CK18, respectively; of the mandibular cysts, 10, 20 and 11 linings were positive for these CKs, respectively. The expression patterns of CK18(+)-CK13(-), CK18(+) CK13(+) and CK18(-)-CK13(+) were observed in 3, 16 and 13 linings of the maxillary cysts and 0, 11 and 9 linings of the mandibular cysts, respectively. In situ hybridization revealed the expression of CK18-mRNA in 9 and 4 linings of 24 maxillary and 13 mandibular cysts examined, respectively. With RT-PCR, we explored that both CK18- and CK13-mRNA were expressed not only in the normal nasal and gingival epithelia but also in the examined maxillary cyst linings although their expression levels differed correlating with the difference in CK staining. CONCLUSION: It is concluded that CK13- and CK18-mRNA are constitutively expressed in columnar and squamous epithelial cells, respectively, and that the variant CK expression patterns with CK18-mRNA expression in maxillary radicular cysts are indicative of the possibility of phenotypic transformation in the cyst linings. PMID- 11936455 TI - Immunohistochemical study on expression of alpha-defensin and beta-defensin-2 in human buccal epithelia with candidiasis. AB - OBJECTIVES AND DESIGN: It has been previously reported that alpha-defensin (HNPs) and beta-defensin-2 (HBD-2) peptides with antifungal and cytotoxic activities can be detected in oral carcinomas and the saliva of patients with oral carcinomas. The present study investigated the presence of HNPs and HBD-2 in oral epithelia with candidiasis. MATERIALS AND METHODS: Tissue sections (4 microm) were prepared from biopsy and surgically removed specimens diagnosed as oral candidiasis (n = 10). The sections were examined immunohistochemically with antibodies directed against HNPs and HBD-2. RESULTS: Tissue sections of oral candidiasis were immunostained with antidefensin antibodies. Neutrophils in the inflamed lamina propria were positively immunostained with anti-HNPs antibody. The cytoplasm of cells in the upper spinous layer, in the lower spinous layer and in the parakeratinized layer of buccal epithelia with candidiasis was immunostained intensely with anti-HBD-2 antibody. In contrast, the expression of HBD-2 in the normal spinous layer was much weaker than that in oral candidiasis. No signals of HNPs were found in normal buccal epithelium. CONCLUSION: Buccal specimens from individuals with oral candidiasis show greater levels of expression of both HNPs and HBD-2. There might be a dual protection manner by defensins against fungal inflammation in infected buccal epithelia locally. Generally, HBD-2 signals have been found everywhere in the buccal epithelium; however, in an infected area, the signal intensity of HBD-2 has increased. HNPs signals have not been found in the normal buccal epithelium; however, HNPs signals have increased when the infection occurred. PMID- 11936456 TI - The use of bone scintigraphy in temporomandibular joint disorders. AB - INTRODUCTION: The use of bone scintigraphy (bone scan) in the diagnosis of temporomandibular joint (TMJ) disease has been infrequent, as compared with traditional radiographic techniques. Bone scans have the potential to detect active bone remodeling whereas corresponding radiographs may be normal or document past structural change in the joint. Traditional radiographic findings and relevant clinical signs and symptoms correlated with bone scans may aid in the diagnosis of TMJ disease and possibly affect treatment and prognosis of individual cases. The use of bone scans as an additional tool in diagnosing TMJ disease was assessed in this series of patients. METHODS: Thirty consecutive subjects with TMJ tenderness were selected for bone scintigraphy using technetium diphosphonate 99 mTc and single photon emission computerized tomography. These subjects received bone scans as well as other selected imaging modalities for diagnostic purposes. RESULTS AND DISCUSSION: The findings on bone scan were evaluated and a change in preliminary clinical diagnosis or treatment was made in 60% of cases because of the findings on bone scintigraphy. Bone scintigraphy may be valuable to assess progress of TMJ inflammation or remodeling, and may affect diagnosis and treatment of patients with TMJ tenderness. PMID- 11936457 TI - Angina bullosa haemorrhagica: presentation of eight new cases and a review of the literature. AB - Angina bullosa haemorrhagica (ABH) describes the acute and sometimes painful onset of oral blood-filled vesicles and bullae not attributable to blood dyscrasia, vesiculo-bullous disorders, systemic diseases or other known causes. The haemorrhagic bullae spontaneously burst after a short time resulting in ragged, often painless, superficial erosions that heal spontaneously within 1 week without scarring. Although the pathogenesis is still unclear, ABH seems to be a multifactorial phenomenon: dental or functional trauma seems to be the major provoking factor. The lesions of ABH can be easily confused with other mucosal diseases. It is important that the presentation of this benign disorder is distinguished from other more serious disorders with similar presenting features. The aim of this paper is to report the clinical features of eight cases of ABH, in an attempt to distinguish ABH from other blistering diseases of oral mucosa and to describe their management. PMID- 11936458 TI - Oral leishmaniasis in a HIV-positive patient. Report of a case involving the palate. AB - Leishmaniasis is a parasitic disease caused by a protozoon (Leishmania), with different clinical forms that are endemic in certain countries. The association of this disease in patients who are seropositive to human immunodeficiency virus (HIV) has recently been described. Leishmaniasis can develop in any stage of HIV infection, although the clinical manifestations - and hence the diagnosis - tend to coincide with the periods of maximum immune depression. We present the case of a HIV-positive, ex-intravenous drug abuser (in stage B2 of the CDC, 1992) with concomitant hepatitis C infection who presented with palatinal pain and bleeding for the past 2 months. Exploration revealed a vegetating tumoration of the hard palate. Hematoxylin-eosin and Giemsa staining of the biopsy confirmed the diagnosis of leishmaniasis. The definitive diagnosis was mucocutaneous leishmaniasis (MCL), for a bone marrow aspirate proved negative, and no further lesions could be established. The patient was treated with meglumine antimoniate (Glucantime), followed by improvement of the lesions. PMID- 11936459 TI - Clinical and radiographic features of a family with autosomal dominant amelogenesis imperfecta with taurodontism. AB - This paper describes the clinical features of a family of four generations with autosomal dominant amelogenesis imperfecta with taurodontism (ADAIT). Considerable variation in phenotype was seen, both between individuals and within the dentition of some individuals. Many of the adults had received extensive dental restorative work. These findings re-enforce previous observations of variable phenotype in this and other forms of the condition and add to the argument for a revision of methods of classification. This history of this large family draws further attention to the restorative demands of this group of dental anomalies and, by their generous co-operation, will prove an invaluable help in the investigation by molecular genetic techniques of this disfiguring condition. PMID- 11936460 TI - Iodine deficiency in Calabria: characterization of endemic goiter and analysis of different indicators of iodine status region-wide. AB - The distribution of goiter prevalence in schoolchildren (no.=13,984, age 6-14 yr), the neonatal TSH results obtained from the congenital hypothyroidism screening program and the urinary iodine excretion values (no.=284) were employed for the assessment of iodine deficiency in Calabria, a Southern Italy region. Data were collected during the years 1990-1996. In the inland territory, goiter prevalence ranged from 19 to 64%. At sea level, there was a great variability of goiter prevalence, with values varying from 5.3 to 25.7%. The analysis of the neonatal hypothyroidism screening program data (no.=21,078) showed a 14.8% frequency of TSH levels >5 microU/ml whole blood in newborns from the inland territory and a 14.1% frequency at sea level. Urinary iodine excretion resulted (mean+/-SD) 53.8+/-43.4 microg/l (range: <20 to 189 microg/l) in the inland territory and 89.6+/-59.8 microg/l (range: 26 to 333 microg/l) at sea level. Median urinary iodine excretion values in 13 villages or small towns of the inland territory ranged from 31 to 57 microg/l. In 2 major towns located at sea level, the median iodine excretion values were 72 microg/l in Crotone main city and 94 microg/l in Reggio Calabria main city. The data indicated that moderate, with pockets of severe iodine deficiency is present in the inland region while iodine supply varies from sufficient to marginally low in the coastal areas. Mild iodine deficiency was found in a major town located at sea level. PMID- 11936461 TI - Relation of three polymorphisms of the CTLA-4 gene in patients with Graves' disease. AB - Graves' disease is an autoimmune disease believed to be caused by a combination of environmental and genetic factors. One of the candidate genes is CTLA-4, a negative regulator of T cell activation. Three polymorphisms of the gene have been described, in the promoter at position -318, at position 49 in exon 1, and an (AT)n repeat within the 3'-untranslated region of exon 4. Many studies describe the association between a polymorphism of the CTLA-4 gene and autoimmune disease. To investigate the association of these CTLA-4 gene polymorphisms with each other, we analyzed the combined frequencies of each polymorphism and calculated the disequilibrium coefficients. We studied DNA samples from 120 Graves' disease (GD) patients and 80 healthy donors (NC). The exon 1 position 49 A/G polymorphism and promoter polymorphism at position -318, were typed using a PCR-restriction fragment length polymorphism method (PCR-RFLP). The polymorphic (AT)n repeat in exon 4 was determined by PCR amplification of genomic DNA, resolution of the amplified products on sequencing gels, and detection by autoradiography. There was a significant difference between GD and NC patients and occurrence of the polymorphism in exon 1 and exon 3, but not for the polymorphism in the promoter region. Furthermore, we found that the genotype with both the G allele in exon 1 and the 106 bp allele of the AT repeat in exon 4 occurred with much higher frequency in GD than NC (p<0.01), and that these polymorphisms are in linkage disequilibrium with each other. These results support the concept that CTLA-4 plays a critical role in the autoimmune process in GD, and that GD depends on multiple genetic susceptibility factors. Because the exon 1 and exon 4 polymorphisms are in strong linkage disequilibrium. It is not possible at this time to determine their unique relation to CTLA-4 function. Studies relating each polymorphism to CTLA4 function are required to determine whether one, or both, polymorphism(s) promote autoimmune disease. PMID- 11936463 TI - Iodine status and goiter prevalence in Turkey before mandatory iodization. AB - Endemic goiter is an important public health problem in Turkey. Legislation for mandatory iodization of household salt was passed in July 1999. Current study is aimed at ascertaining the goiter prevalence and iodine nutrition in school-age children (SAC) living in known endemic areas of Turkey. Sonographic thyroid volumes (STV) and urinary iodine concentrations (UIC) of 5,948 SAC from 20 cities were measured between 1997-1999. STV of 31.8% of the SAC examined stayed above the upper-normal limits for the same age and gender recommended by the World Health Organization (WHO). Goiter prevalence ranged between 5 to 56% and median UIC ranged between 14 to 78 microg/l, indicating severe to moderate iodine deficiency (ID) in 14 and mild ID in 6 of the cities surveyed. Neither of the cities was found to have sufficient median UIC levels. The current study shows that endemic goiter is an important public health problem and iodine nutrition is inadequate nationwide. It also provides reliable scientific evidence and shows the need for a controlled and effective iodine supplementation program nationwide. Mandatory iodization of household salt seems to be the essential measure taken for the moment, additional measures may be needed in the near future. PMID- 11936462 TI - Activity of GH/IGF-I axis in trauma and septic patients during artificial nutrition: different behavior patterns? AB - The aim of this study was to compare several parameters of GH/IGF-I axis activity in septic and trauma patients during Intensive Care Unit (ICU) stay. To this goal, 13 patients with sepsis (SEP) and 16 with trauma (TRA) were studied. Thirty three adult subjects (AS) were studied as controls. Serum IGF-I and -II, IGFBP-1, -2 and -3, GH and GHBP levels were studied on day 1, 3, 5 and 7 after ICU admission, during comparable artificial nutrition in SEP and TRA and basally in AS. In 5 patients with SEP and 6 with TRA, the GH response to GHRH was evaluated on day 3. On ICU day 1, IGF-I and -II and IGFBP-3 in SEP were lower (p<0.05) than in TRA which, in turn, were lower (p<0.01) than in AS. IGF-I increased (p<0.05) both in SEP and TRA, but, on ICU day 7, those in SEP persisted lower than in TRA, which became similar to those in AS. IGF-II levels increased (p<0.05) in SEP only, persisting lower (p<0.05) than in TRA. On ICU day 1, GH in SEP and TRA were similar and did not vary until day 7, overlapping those in AS. The GH response to GHRH in SEP and TRA was similar and lower (p<0.01) than in AS. These findings indicate that IGF-I activity is impaired more in septic than in trauma patients. Reduced IGF-I activity probably reflects peripheral GH resistance though basal and GHRH-induced GH levels were not increased in these conditions. PMID- 11936464 TI - Central and peripheral glucocorticoid receptor function in abdominal obesity. AB - Abdominal obesity seems to be associated with a moderately deranged feedback regulation of the hypothalamic-pituitary-adrenal (HPA) axis where central glucocorticoid receptors (GR) are involved. Therefore, functions of central and peripheral GR were compared in this study. Furthermore, since trinucleotide repeats in early exons of steroid hormone receptor genes influence transcription, and therefore may influence receptor density, this was also studied. Ten middle aged men, 5 with abdominal obesity and 5 controls, were studied. The suppression of dexamethasone (dex) on serum cortisol was used in dose-response tests to assess the function of central GR. Abdominal adipose tissue biopsies were incubated and exposed to cortisol in different concentrations, and the function of the peripheral GR assayed as induction of lipoprotein lipase (LPL) activity. Aberrant expansion of exonic trinucleotide repeats in the first coding exon of the GR gene was studied by sequencing of genomic DNA. Results showed that men with abdominal obesity showed less inhibition of serum cortisol by dex, particularly at lower concentrations, while in the controls cortisol secretion was inhibited in an apparent dose-response manner. LPL activity in adipose tissue was lower in abdominal obese men than in controls. However, the sensitivity to cortisol was not different between the groups. There was no evidence for expansion of trinucleotide repeats. These results suggest that the central GR and the peripheral GR in adipose tissue exhibit functional differences in abdominal obesity. PMID- 11936465 TI - Abnormalities of hypothalamic-pituitary-thyroid axis in patients with primary empty sella. AB - Primary empty sella (PES) is a very frequent neuroradiological finding in the general population, that can induce hypopituitarism. Some studies focused on the association of PES with GH deficiency (GHD) or hypogonadotropic hypogonadism (HH), while data regarding the involvement of hypothalamic-pituitary-thyroid (HPT) axis, despite sporadic reports of central hypothyroidism, or the occurrence of hypoadrenalism (HA) are scanty. In this study, thyroid function and TSH response to exogenous TRH injection (TRH/TSH) were investigated in 43 patients [10 men and 33 women; aged (mean +/- SD), 48+/-12 yr] with PES: 22 patients had total and 21 partial PES. Forty healthy subjects (9 men and 31 women; aged 46+/ 12 yr) were enrolled as a control group. Central hypothyroidism was found only in 2/43 cases, whereas one patient showed primary hypothyroidism. In euthyroid patients, mean serum TSH levels were significantly lower than controls (TSH: 1.0+/-0.7 vs 1.4+/-0.6 mU/l, p<0.01) and 79% of them showed abnormal TRH/TSH responses (TRH test was performed in 34 euthyroid patients: 17 cases with total and 17 cases with partial PES), but mean serum free T4 (FT4) and free T3 (FT3) values were not significantly lower than controls (FT4: 15.9+/-0.4 vs 15.0+/-2.1 pmol/l, p=NS; FT3: 5.3+/-1.2 vs 5.8+/-1.5 pmol/l, p=NS). Moreover, no significant differences were evident in mean serum TSH, FT4 and FT3 between patients with total and partial PES (TSH: 1.1+/-0.7 vs 0.9+/-0.8 mU/l, p=NS; FT4: 16.3+/-2.6 vs 15.7+/-2.2 pmol/l, p=NS; FT3: 5.4+/-1.3 vs 5.2+/-0.8 pmol/l, p=NS) and the TRH/TSH peak was impaired or exaggerated/delayed in 9 and 3 patients with total and in 12 and 3 cases with partial PES. No significant differences in the prevalence of abnormal TRH/TSH responsiveness were found between patients with partial or total PES (chi2=1.6, p=NS). Other impairment of pituitary function was detected in 23/43 patients: GHD was present in 15 cases, HH in 11 and central HA in 5 patients. Isolated or combined hypopituitarism was present in 17 and in 6 patients, respectively. In conclusion, pituitary dysfunction is very frequent in patients with PES, but central hypothyroidism occurs rarely. The entity of arachnoid herniation into the sellar fossa does not play a significant role on the degree of HPT axis dysfunction. PMID- 11936466 TI - Prevalence of thyroid diseases in patients with acromegaly: results of an Italian multi-center study. AB - Acromegaly is frequently associated with the presence of thyroid disorders, however the exact prevalence is still controversial. An Italian multicenter study was performed on 258 patients with active acromegaly (high levels of IGF-I and lack of suppression of serum GH levels below 2 microg/l after an OGTT). The control group was represented by 150 patients affected by non-functioning and PRL secreting pituitary adenomas. Two hundred and two out of 258 acromegalic patients (78%) were affected by thyroid disorders with a significantly higher prevalence with respect to the control group (27%, p<0.0001). One hundred and three patients presented (39.9%) non-toxic nodular goiter, 46 (17.8%) non-toxic diffuse goiter, 37 (14.3%) toxic nodular goiter, 1 toxic diffuse goiter (0.4%), 12 (4.6%) Hashimoto's thyroiditis, 3 (1.2%) thyroid cancer. Two patients presented a co secreting TSH pituitary adenoma. Thirty-six patients had been previously treated for various thyroid abnormalities. Among the 222 acromegalic patients never treated for thyroid disorders thyroid ultrasonography was performed on 194 subjects. Thyroid volume in patients with thyroid abnormalities was 28+/-17.5 ml (median 23) while it was 10.8+/-3.6 ml (median 10) in patients without thyroid disorders (p<0.0001). Thyroid volume was correlated with the estimated duration of acromegaly (r=0.7, p<000.1), but not with age or with serum GH, IGF-I and TSH concentrations. Thyroid volume was higher in acromegalic patients than in the above control population (23.5+/-16.9 ml vs 13.9+/-12.8 ml, p<0.0001). In 62 acromegalic patients 101 fine-needle biopsies of thyroid nodules were performed; 7 nodules were suspicious and the patients were submitted to thyroid surgery: papillary thyroid carcinoma was found in 3 patients. In conclusion, in a large series of acromegalic patients an increased prevalence of thyroid disorders (78%), particularly non-toxic nodular goiter, has been observed. Thyroid volume, evaluated by ultrasonography, was correlated to the estimated duration of acromegaly. Finally, the prevalence of thyroid carcinoma was slightly increased than in the general population. PMID- 11936467 TI - Apathetic Graves' disease and acquired hemophilia due to factor VIIIc antibody. AB - Acquired hemophilia due to autoantibody to Factor VIII coagulant (Factor VIIIc) is a rare event which may be observed in patients with different autoimmune diseases. To our knowledge, this association has been reported only once in patients with autoimmune thyroid disease. Here we describe a patient presenting with a severe hemorrhagic disorder due to Factor VIIIc antibody in whom biochemical screening for thyroid diseases led to a diagnosis of hyperthyroid Graves' disease not associated to overt clinical features. This case underlines the importance of carrying out a complete screening for autoimmunity, including thyroid autoimmune disease, in all patients with apparently isolated serum Factor VIIIc inhibitors. PMID- 11936468 TI - Aggressive behavior of papillary microcarcinoma in a patient with Graves' disease initially presenting as cystic neck mass. AB - Prognosis of papillary carcinoma of the thyroid associated with Graves' disease is controversial; nevertheless, tumors smaller than 1 cm (microcarcinoma) are usually considered to render a good prognosis. We describe a patient with Graves' disease who developed a lateral cystic neck mass that was later confirmed to be a metastatic lymph node from papillary thyroid microcarcinoma. Contrary to having a good prognosis with a microcarcinoma, our patient developed bilateral lung metastases. The possible role of thyroid stimulating immunoglobulins in the aggressive course of the tumor is discussed. PMID- 11936469 TI - A case of severe hypertension caused by ACTH-independent macronodular adrenal hyperplasia. AB - This report describes a rare case of ACTH-independent macronodular adrenal hyperplasia (AIMAH) arisen with symptomatic severe hypertension and hypokaliemia. A 55-year-old man was admitted to hospital with a clinical picture characterized by several episodes of transient ischemic attacks (TIA) and right hemiplegia, related to severe arterial hypertension. Laboratory tests showed urinary levels of catecholamines, metanephrines and vanillylmandelic acid (VMA) in normal range; high urinary free cortisol excretion, elevated serum cortisol with loss of the circadian rhythm and low ACTH plasma levels. ACTH failed to respond to CRH administration. Serum cortisol levels were not modified after high doses of dexamethasone. MRI showed bilateral macronodular hyperplasia of adrenal glands, whereas pituitary-MRI did not show tumoral lesions. Therefore, ACTH-independent macronodular hyperplasia was suspected. Though obese, the patient had no typical Cushing habit, and symptomatic hypertension with hypokaliemia was the only clinical evidence for this rare kind of Cushing's syndrome. After obtaining a satisfactory control of blood pressure, the patient was successfully submitted to laparoscopic bilateral adrenalectomy and underwent complete clinical remission. The histology showed adrenal macronodular hyperplasia. During the twenty-four month follow-up, the patient had no further transient ischemic attacks or need of glucocorticoid replacement therapy and withdrew the antihypertensive drugs. PMID- 11936470 TI - Study of the family of a patient with male-limited precocious puberty (MPP) due to T1193C transition in exon 11 of LH receptor gene. AB - Molecular diagnostics of the LHR gene was conducted in a 5-year-old boy with clinical symptoms and hormonal profile typical of precocious puberty. His parents and 4 sisters were also diagnosed. Single-strand conformation polymorphism analysis under temperature gradient conditions (Multitemperature SSCP) of 3 overlapping fragments of exon 11 of LHR gene revealed a mutation in the fragment spanning nucleotides 1072 to 1804. This mutation was found in the patient, in his mother and in his 4 sisters, and was confirmed by digestion with the use of restriction enzyme Bbr Cl. Direct sequencing revealed a heterozygous T1193C transition in the DNA fragment of the patient and in one of the alleles of his mother's and sister's DNA. This mutation causes Met398Thr substitution in the second transmembrane helix and results in a constitutive activation of LH receptor. This is the second identical mutation detected in Poland and one of the 7 identified so far in the world population. PMID- 11936471 TI - Lingual thyroid and hyperthyroidism: a new case and review of the literature. AB - Lingual thyroid is the result of a defective migration of the thyroid anlage occurring between the 3rd and 7th week of gestation. Whereas mutations in the transcription factor-2 (TTF-2) and PAX8 and in the TSH receptor genes (TSH-R) have been reported in a minority of patients with thyroid dysgenesis, the etiopathogeny of the majority of cases, and in particular of thyroid ectopy, remains unclear. The majority of patients with thyroid ectopy are asymptomatic, but obstructive symptoms as well as hypothyroidism have been observed. Hyperthyroidism is an exceptionally rare finding. To our knowledge, only 2 cases have been reported in the literature to date. Herein, we describe an unusual case of thyrotoxicosis related to a nodular lesion in a lingual thyroid. Treatment consisted in restoration of a euthyroid state with thionamide followed by surgical removal of the ectopic gland. The underlying molecular cause of the ectopic lingual thyroid and the toxic adenoma in this case could not be identified. We speculate that abnormally early differentiation of the thyroid gland could interfere with the migration process, a hypothesis yet to be confirmed. PMID- 11936473 TI - Different intrathyroid expression of intercellular adhesion molecule-1 (ICAM-1) in Hashimoto's thyroiditis and Graves' disease: analysis at mRNA level and association with B7.1 costimulatory molecule. AB - Cultured thyroid epithelial cells can be induced to express intercellular adhesion molecule-1 (ICAM-1, or CD54). However, constitutive follicular expression of ICAM-1 has been reported only in thyroid autoimmunity. We evaluated the expression of ICAM-1 mRNA and protein on thyroid tissue from different autoimmune thyroid diseases, and its relationship with other immunologically relevant surface markers, namely costimulatory molecules of B7 family. Thyroid tissue sections were obtained by surgically removed thyroid glands from 6 patients with Hashimoto's thyroiditis (HT), 6 with Graves' disease (GD) and 3 with multinodular nontoxic goiter. We used in situ hybridization to localize ICAM 1 mRNA, and immunohistochemical analysis by alkaline phosphatase anti-alkaline phosphatase (APAAP) method. We showed a clear hybridization pattern, localized in follicular cells, in sections of glands with HT. The hybridization pattern was far less pronounced in GD: no staining was apparent on follicular cells. These results were strictly consistent with those obtained by means of immunohistochemistry. Moreover, double-staining experiments demonstrated colocalization of ICAM-1 and B7.1 molecules in HT, whereas no B7.1 expression was observed in Graves' or in non-autoimmune thyroid diseases. These data agree with the hypothesis of distinct immunoregulatory phenomena and effector mechanisms in the 2 main autoimmune thyroid diseases. PMID- 11936474 TI - Mediterranean diet and prevention of coronary heart disease. PMID- 11936475 TI - Woman of Naples countryside. PMID- 11936476 TI - Primary hyperparathyroidism is associated with an impaired secretion of growth hormone but not of the other anterior pituitary hormones. AB - We have recently reported that GH secretion is impaired in primary hyperparathyroidism (PHP). No systematic assessment of the whole anterior pituitary function in PHP is available. In this study, anterior pituitary function was evaluated in basal and stimulated conditions in a series of 12 consecutive women with PHP. Serum GH secretion was decreased in 9 of 12 PHP patients (75%) confirming our previous results in different series of PHP patients. Instead, at variance, secretion of all the other anterior pituitary hormones was normal. Thus, PHP is associated with an impaired secretion of GH, but not of the other anterior pituitary hormones. The reason why only GH secretion is affected in PHP is unknown. PMID- 11936477 TI - US demonstration of a thrombosed persistent median artery in carpal tunnel syndrome. AB - Median artery of the forearm and wrist is not very frequently observed because it normally involutes before birth. Only a few cases of persistent median artery thrombosis associated with compression of the median nerve in the carpal tunnel have been reported. In these cases symptoms arise suddenly and surgery consists of the excision of the thrombosed arterial branch. In cases of large persistent unthrombosed median artery associated with carpal tunnel syndrome (CTS), excision of the unthrombosed median artery is not indicated because it may sometimes substantially contribute to the circulation of the hand. We report the case of a 39-year-old man with CTS associated with a thrombosis of a persistent median artery detected by high-resolution US and Doppler ultrasound. US can be also useful to exclude other causes of CTS such as tenosynovitis of the flexor tendons, ganglion cyst, musculotendinous variants, and various soft tissue tumors. PMID- 11936472 TI - Action of thyroid hormone in brain. AB - Among the most critical actions of thyroid hormone in man and other mammals are those exerted on brain development. Severe hypothyroidism during the neonatal period leads to structural alterations, including hypomyelination and defects of cell migration and differentiation, with long-lasting, irreversible effects on behavior and performance. A complex regulatory mechanism operates in brain involving regulation of the concentration of the active hormone, T3, and the control of gene expression. Most brain T3 is formed locally from its precursor, T4, by the action of type II deiodinase which is expressed in glial cells, tanycytes, and astrocytes. Type III deiodinase (DIII) is also involved in the regulation of T3 concentrations, especially during the embryonic and early post natal periods. DIII is expressed in neurons and degrades T4 and T3 to inactive metabolites. The action of T3 is mediated through nuclear receptors, which are expressed mainly in neurons. The receptors are ligand-modulated transcription factors, and a number of genes have been identified as regulated by thyroid hormone in brain. The regulated genes encode proteins of myelin, mitochondria, neurotrophins and their receptors, cytoskeleton, transcription factors, splicing regulators, cell matrix proteins, adhesion molecules, and proteins involved in intracellular signaling pathways. The role of thyroid hormone is to accelerate changes of gene expression that take place during development. Surprisingly, null mutant mice for the T3 receptors show almost no signs of central nervous system involvement, in contrast with the severe effects of hypothyroidism. The resolution of this paradox is essential to understand the role of thyroid hormone and its receptors in brain development and function. PMID- 11936478 TI - CT findings in eosinophilic enterocolitis with predominantly serosal and muscular bowel wall infiltration. AB - A 44-year-old female presented with tenderness of her abdomen, vomiting, intestinal obstruction, hypoalbuminemia and blood eosinophilia. Gastroscopy was normal and colonoscopic biopsies showed only non-specific inflammation of the colonic mucosa and submucosa. CT revealed large amounts of ascites and bilateral pleural effusions but eosinophil counts in the ascites were normal. At CT the jejunum was dilated and showed marked prominence of the valvulae whereas the ileum and the colon presented with a diffuse and hypoattenuating bowel wall thickening. The bowel wall thickening was most pronounced in the colon which especially showed also an impressive thickening and hyperenhancement mainly of its outer bowel wall layers. Parasitic infection could be excluded as well as a specific allergic response. In context with the known blood eosinophilia the diagnosis of an eosinophilic enterocolitis was suspected already by CT but finally only surgical full thickness biopsies could confirm the rare diagnosis of an eosinophilic enterocolitis with predominantly serosal and muscular bowel wall infiltration. PMID- 11936479 TI - Acute obstructive gastric volvulus diagnosed by helical CT. PMID- 11936480 TI - Intraductal papillary mucinous tumor of the pancreas demonstrated by MR cholangiopancreatography. PMID- 11936481 TI - Randomized, double-blind, placebo-controlled study of subcutaneous interferon beta-1a in relapsing-remitting multiple sclerosis: a categorical disability trend analysis. AB - The treatment effects of recent immunomodulatory therapies on disease progression in relapsing-remitting multiple sclerosis (MS) have been mostly established from 'confirmed progression' endpoints. However, the reliability of this outcome measure is poor and a significant proportion of patients may be erroneously classified. We previously proposed the area under disability/time curves to quantify in-trial disability changes, but although these have advantages, they lack information on the direction of change. We have therefore performed disease trend analyses and categorical classifications using serial Expanded Disability Status Scale (EDSS) scores from the 533 complete datasets in a double-blind, randomized, placebo-controlled, phase III trial of subcutaneous interferon beta 1a (IFNbeta-1a) (PRISMS study). We found significant treatment benefits for IFNbeta-1a on in-trial disability course (P=0.002). Therapeutic advantages remained when relapse-related assessments were excluded (P=0.018). Post hoc analyses demonstrated that IFNbeta-1a was mainly effective in both increasing the proportion of patients with a 'stable' course and reducing those with prolonged, disabling deteriorations. Baseline disease duration and EDSS levels, but not MRI lesion load, predicted the subsequent disability trends. Mean 'numbers needed to treat' (NNTs) to obtain preferred disability courses were reduced in patients with shorter disease duration. These results have important implications for the targeting of immunomodulatory therapies in MS. PMID- 11936482 TI - Both paracetamol and ibuprofen are equally effective in managing flu-like symptoms in relapsing-remitting multiple sclerosis patients during interferon beta-1a (AVONEX) therapy. AB - Interferon beta-1a is an established therapy for patients with relapsing remitting multiple sclerosis (MS). Adverse effects in the first weeks of treatment are common. This open-label, multicenter, randomized, prospective study compared treatment of flu-like symptoms (FLS) with paracetamol versus ibuprofen administered 48 h within interferon injection. The percentage of patients with FLS was comparable between both treatment groups and improved during the course of the study (baseline: paracetamol 92%, ibuprofen 90%; week 12: paracetamol 60%, ibuprofen 57%). More than 75% of patients receiving either paracetamol or ibuprofen reported no or only mild impairment of daily activities. There was no significant difference in general satisfaction or incidence of additional symptoms (weakness, nausea, headache; paracetamol 84.6% patients, ibuprofen 86.0% patients) between the two groups. A significant overall improvement from baseline to week 12 was observed for all parameters studied (paracetamol and ibuprofen groups were pooled). These results indicate that neither the paracetamol nor the ibuprofen treatment regimen is better. PMID- 11936484 TI - Use of interferon beta in multiple sclerosis: rationale for early treatment and evidence for dose- and frequency-dependent effects on clinical response. AB - The current approach to the use of interferon (IFN) beta in the treatment of multiple sclerosis (MS) is, in general, conservative. However, recent findings about early events in MS and data on dose-response relationships with IFN beta indicate that such an approach may be suboptimal. Four lines of evidence suggest that delays in the initiation of therapy with IFN beta may be detrimental: 1) axonal damage secondary to inflammation starts very early in the course of MS; 2) pathological events occurring early in MS are predictive of the future course of the disease; 3) inflammatory activity in relapsing MS is not confined to episodes of clinical impairment, but often starts before the first such episode and generally continues during remissions; and 4) the immune-mediated activity that underlies MS may become more difficult to control as the disease progresses. An early treatment strategy is also supported by data from two recently published clinical studies. In addition, preclinical and clinical results suggest that the beneficial effects of IFN may be dose- and frequency-dependent. Taken together, these findings indicate that treatment with IFN beta should be started as early as possible in the course of MS, and suggest that, in order to maximize patent benefit, the highest possible dose of IFN beta should be chosen. PMID- 11936483 TI - International consensus statement on the use of disease-modifying agents in multiple sclerosis. AB - OBJECTIVE: To provide recommendations on the use of disease-modifying agents in the management of multiple sclerosis (MS) and to ensure that treatment will be available to those patients who may benefit. METHODS: An initial draft of the consensus statement was prepared by the Steering Committee and amended in the light of written comments from a group of MS specialists. At a subsequent workshop, the wording of the consensus statement was discussed, modified if necessary, and the participants indicated their level of support using an electronic voting system. A new draft of the statement was then sent to a much larger group of international opinion leaders in MS for further comment. RESULTS: A number of statements were agreed, which outline the criteria for consideration of disease-modifying therapy for MS and recommendations for treatment. Each statement was accepted completely, or with only minor reservations by 95% or more of those present at the workshop. CONCLUSIONS: Periodic reviews and modifications to the statement will be required, as new approaches to the treatment of MS and other therapeutic agents become available. PMID- 11936485 TI - A pilot study of recombinant insulin-like growth factor-1 in seven multiple sderosis patients. AB - The purpose of this open-label, crossover study was to determine the safety and efficacy of recombinant insulin-like growth factor-1 (rhIGF-1) using magnetic resonance imaging (MRI) and clinical measures of disease activity in seven multiple sderosis (MS) patients. Monthly clinical and MPI examinations were performed during a 24-week baseline and a 24-week treatment period with rhIGF-1. The primary outcome measure was contrast enhancing lesion (CEL) frequency on treatment compared to baseline. Secondary outcome measures included dinical and MRI measures of disease activity including: white matter lesion load (WMLL), magnetization transfer ratio (MTR), T1-Hypointensity volume, cervical spine cross sectional area and proton magnetic resonance spectroscopic (MRS) imaging for determining regional metabolite ratios. rhIGF-1 (Cephalon) was administered at a dose of 50 mg subcutaneously twice a day for 6 months. rhIGF-1 was safe and well tolerated with no severe adverse reactions. There was no significant difference between baseline and treatment periods for any MRI or clinical measures of disease activity. Although rhIGF-1 did not alter the course of disease in this small cohort of MS patients, the drug was well tolerated. Further studies using rhIGF-1 alone or in combination with other therapies may be of value because of the proposed mechanism of action of this growth factor on the oligodendrocyte and remyelination. PMID- 11936486 TI - A double-blind pilot study of the effect of Prokarin on fatigue in multiple sclerosis. AB - In this 12-week study with 29 subjects, the effect of Prokarin (n=22), a proprietary blend of histamine and caffeine, was compared to placebo group (n=7) for the following outcomes: 1) fatigue as measured by the Modified Fatigue Impact Scale (MFIS); 2) lower limb function as measured by timed walk test; 3) upper limb function as measured by the pegboard test; 4) cognitive function as measured by the Paced Auditory Serial Additions Test (PASAT); 5) serum caffeine level; 6) change in brain chemistry as measured by quantitative magnetic resonance spectroscopy assay of N-acetyl aspartate (NAA); and 7) safety as measured by routine blood chemistry, TSH and urinalysis. Data were acquired at baseline, 4, 8 and 12 weeks. The Prokarin group MFIS mean was significantly different rom the mean of the placebo group at 12 weeks (df=24, t=2.08, P=<0.02), with respective means of 37.40, SD=15.18, for the Prokarin group and 53.2, SD=11.39 for the controls. For the secondary endpoints (PASAT, 25 foot timed walk, peg test, and magnetic resonance spectroscopy [MRS]), there were no significant differences between the Prokarin-treated group and the placebo group. However, there were significant improvements within the Prokarin group for each of these measures for the pre- versus posttreatment comparison at 12 weeks. Serum caffeine data indicated that caffeine exerted no independent effect on performance. No laboratory abnormalities were seen, and the treatment was well tolerated. CONCLUSION: There was a modest-size statistical effect of Prokarin on fatigue in multiple sderosis (MS) compared with the placebo group. A larger trial is warranted, based on this pilot study. PMID- 11936487 TI - The interdisciplinary approach to the treatment of multiple sclerosis patients in Italy: an aspiration or a reality? AB - The aim of the study was to assess whether the components necessary for an interdisciplinary approach to the treatment of multiple sclerosis (MS) patients exist in Italian MS clinics and to identify factors that make the difference between reality and aspiration for this type of approach. One hundred two out of 147 neurology departments with MS clinics throughout Italy compiled a questionnaire about characteristics and organizational structure of the clinic, type and number of professionals and resource needs. Eight hundred twenty-three healthcare professionals are currently working in MS clinics: 50% (412) neurologists, 28.7% (236) nurses, 15.1% (124) physical therapists, 3.4% (28) psychologists and 2.8% (23) social workers. Neurologists are evenly distributed between northern and southern parts of the country even though there are nearly double the number of patients followed in northern dinics compared to those in the south. Physicians reported themselves as most in need of continuing education, twice that which the same physicians reported for any other professional. The study has identified issues that contribute to the difficulty in developing and applying an interdisciplinary approach to providing care and services to Italian MS patients. Several factors have been delineated that require significant reshaping in order for this approach to begin to develop. PMID- 11936488 TI - The role of MRI as a surrogate outcome measure in multiple sclerosis. AB - The need for more specific and more sensitive outcome measures for use in testing new therapies in multiple sderosis (MS) is generally accepted. This need has been accentuated by the realization that the ability to conduct large placebo controlled trials will be limited in the future. From the first use of magnetic resonance imaging (MRI) to study MS, the ability of this imaging technique to identify areas of the central nervous system damage by the disease process in MS has been impressive. Thus, the possibility that MRI could serve as a surrogate outcome measure in clinical trials in MS has been attractive. The use of MRI as a surrogate outcome measure has been examined by an international group of investigators with expertise in clinical aspects of MS, the use of MRI in MS, and in experimental therapeutics. The group agreed that MRI does not represent a validated surrogate in any clinical form of MS. It was also agreed, however, that MRI does provide a reflection of the underlying pathology in the disease, but no single MRI measurement in isolation was seen as sufficient to monitor disease. The use for multiple imaging techniques, especially new, emerging techniques that may better reflect the underlying pathology, was seen as particularly important in monitoring studies of patients with either secondary or primary progressive MS. The choice of MRI techniques used to monitor new therapies needs to be consistent with the proposed mechanisms of the new therapy and phase of the disease. It was also noted, however, that additional validation is required for nonconventional imaging techniques. Finally, the participants noted that clinical trials using MRI as a primary outcome measure may fail to fully identify the effects of the therapy on dinical measures and that the risk and cost-benefit ratio of the treatment might be unresolved. Thus, before MRI is used as a primary outcome measure, new approaches to trial design must be given careful consideration. PMID- 11936489 TI - Contribution of cervical cord MRI and brain magnetization transfer imaging to the assessment of individual patients with multiple sclerosis: a preliminary study. AB - This study was performed to assess how established diagnostic criteria for brain magnetic resonance imaging (MRI) interpretation in cases of suspected multiple sclerosis (MS) (Barkhofs criteria) would perform in the distinction of MS from other diseases and whether other MR techniques (cervical cord imaging and brain magnetization transfer imaging [MTI]), might help in the diagnostic work-up of these patients. We retrospectively identified 64 MS and 59 non-MS patients. The latter group included patients with systemic immune-mediated disorders (SID; n=44) and migraine (n=15). All patients had undergone MRI scans of the brain (dual echo and MTI) and of the cervical cord (fast short-tau inversion recovery). The number and location of brain T2-hyperintense lesions and the number and size of cervical cord lesions were assessed. Brain images were also postprocessed to quantify the total lesion volumes (TLV) and to create histograms of magnetization transfer ratio (MTR) values from the whole of the brain tissue. Barkhofs criteria correctly classified 108/123 patients, thus showing an accuracy of 87.8%. "False negative" MS patients were 13, while 2 patients with systemic lupus erythematosus (SLE) were considered as "false positives". Using brain T2 TLV, nine of the "false negative" patients were correctly classified. Correct classification of 10 MS patients and both the SLE patients was possible based upon the presence or absence of one cervical cord lesion. Two MS patients with negative Barkhofs criteria and no cervical cord lesions were correctly classified based on their brain MTR values. Overall, only one MS patient could not be correctly classified by any of the assessed MR quantities. These preliminary data support a more extensive use of cervical cord MRI and brain MTI to differentiate between MS and other disorders in case of incondusive findings on T2-weighted MRI scans of the brain. PMID- 11936491 TI - The emerging role of MRI in multiple sclerosis and the new diagnostic criteria. PMID- 11936490 TI - Incidence of enhancement of the trigeminal nerve on MRI in patients with multiple sclerosis. AB - The purpose of this study is to review the incidence of contrast enhancement of the trigeminal nerve on magnetic resonance imaging (MRI) in a large cohort of patients with multiple sclerosis (MS). MRI scans of 851 MS patients participating in a phase III clinical trial were reviewed for the presence of gadolinium enhancement on postcontrast T1-weighted images. If enhancement was present, it was documented whether this involved only the proximal or also the distal part of the fifth nerve, and whether the abnormality was unilateral or bilateral. In 24 (2.8%) patients, enhancement of the trigeminal nerve was observed, and was bilateral in 16 (66.7%) of those. In 19 (79.2%) patients with abnormal nerves, enhancement extended to the distal part of the trigeminal nerve (into Meckel's cave). The results of this study indicate a high, probably clinically silent, incidence of trigeminal nerve demyelination in MS and frequent involvement of the peripheral type of myelin in MS. PMID- 11936492 TI - The natural history of secondary progressive multiple sclerosis: observations from the London study group. PMID- 11936493 TI - Immunological aspects of secondary progressive multiple sclerosis. PMID- 11936494 TI - MRI aspects of secondary progressive multiple sclerosis. PMID- 11936495 TI - Therapeutic approaches to secondary progressive multiple sclerosis. PMID- 11936496 TI - A light and electron microscope study of Sarcocystis mitrani (sp. nov.) infecting the skink Scincus mitranus in the central region of Saudi Arabia. AB - The prevalence of Sarcocystis infection among skinks, Scincus mitranus, was studied for the first time. Grossly macroscopic sarcocysts were found to infect the skeletal muscles of the skink (infection rate: 4.16%). Fecal examination for the presence of sporocysts was negative in this study. Sarcocysts were studied using light and transmission electron microscopes. Mature sarcocysts measuring 0.05-0.3 x 0.5-1.8 mm (mean 0.1 5x 1.2 mm) were observed. The characteristic primary cyst wall, with long, finger-like, non-branched and non-stalked protrusions, is described. The ground substance gives rise to numerous thick septa dividing the interior of the cyst into chamber-like compartments. Zoites, including metrocytes and merozoites, were found to have the main architecture of Apicomplexa. Peculiarities of these elements and the importance of the primary cyst-wall ultrastructure for identification and specification of Sarcocystis are discussed. Secondary cyst wall was completely absent. Alterations in the infected host cell were observed. PMID- 11936497 TI - Human cystic echinococcosis in the West Bank of Palestine: surgical incidence and seroepidemiological study. AB - The surgical incidence of cystic echinococcosis (CE) due to Echinococcus granulosus was investigated in hospitals of the West Bank, Palestinian Authority between January 1990 and December 1997. Serum samples from school-children in Yata town, which showed the highest surgical incidence, were tested for anti hydatid antibodies. A total of 390 surgically confirmed cases were recorded throughout the 8-year period, with an overall mean annual surgical incidence (MASI) of 3.1 per 100,000. A high MASI of 4.9, 5.0 and 5.1 per 100,000 was found in Hebron, Jericho and Bethlehem Governorates, respectively. Yata town, Hebron governorate, showed the highest MASI, at 16.8 per 100,000. The highest incidence was found in age groups 11-20 and 21-30 years, at 27.4% and 21.5% of the total number of cases. While there was no significant gender difference in the number of cases in the age groups of 20 years or less, the male to female case ratio was 1:3.2-4.1 in the older age groups. The liver was the most common site of hydatid cysts in 69.9% of cases. Lung cysts were predominant in younger age groups (20 years or less). The seropositivity for CE in the school-children of Yata was 2.4% and 2.1% using enzyme-linked immunosorbent assay and the indirect haemagglutination test, respectively. CE is a significant endemic disease throughout the West Bank. The disease is acquired early in life and is more prevalent among females than males. Behaviour and life-style favour the spread of the disease. PMID- 11936498 TI - Recognition of plasmodium falciparum proteins by mannan-binding lectin, a component of the human innate immune system. AB - The mannan-binding lectin (MBL) is a serum protein, which is involved in the immune defence against viruses, bacteria and parasites. Children who have mutations in the MBL gene that lead to a MBL deficiency are more susceptible to infectious diseases and are more likely to suffer from severe malaria. In this report we investigate the interaction between MBL and the proteins of red blood cells infected with the parasite Plasmodium falciparum. Protein extracts were separated on MBL-sepharose columns. After the elution of bound material, the proteins were detected either by Western blot with human antibodies, or radioactive labelling with 35S-methionine or 3H-glucosamine. MBL recognises proteins of P. falciparum-infected erythrocytes that are immunogenic in humans, parasite-derived and glycosylated. Whether the proteins identified in the different assays are identical remains to be explored. MBL added to in vitro cultures of P. falciparum, however, does not inhibit parasite growth. The positive effect of MBL in the blood of malaria patients could be caused by detoxification of parasite products. PMID- 11936499 TI - Lectin-binding by sporozoites of Elmeria tenella. AB - Sporozoites of Eimeria tenella were reacted in vitro with 19 different lectins characterized with a variety of carbohydrate-binding properties. Nine lectins caused sporozoite agglutination, which was inhibited by the specific carbohydrates mannose, sialic acid, melibiose, D-galactose, or D-galNAc. When intact live or fixed whole sporozoites were reacted with fluorescein isothiocyanate-conjugated lectins, another nine lectins bound to sporozoites, giving weak to strong fluorescence but not agglutination. Of these, all nine lectins bound to surface sites, but four also bound to the refractile body. Two of the agglutinating lectins also bound to intracellular organelles of air-dried sporozoites. SDS-PAGE analysis showed that biotinylated lectins bound a wide variety of parasite proteins. Lectins with similar carbohydrate specificities had some similarity in binding patterns of parasite proteins, as well as marked differences. In a few cases lectins with different carbohydrate specificities bound common protein bands. Only one lectin (Dolichos biflorus) showed no evidence of binding to whole sporozoites, organelles, or proteins. PMID- 11936500 TI - DNA polymorphism in north indian isolates of Entamoeba histolytica detected by PCR fingerprinting. AB - Entamoeba histolytica infection in humans is still one of the leading causes of parasitic diseases among the developing countries, including India. It therefore becomes extremely important to characterize this parasite with the aim of preventing and controlling the amoebiasis which it causes. The present study describes for the first time the ability of random amplified polymorphic DNA (RAPD)-PCR fingerprinting strategies to subtype amoebae and to detect considerable genetic variability present among the various North Indian isolates of E. histolytica studied. The number of rapdemes generated by the application of arbitrary primers in RAPD-PCR showed a significant variation in these isolates, both in the size and number of bands, thus revealing the presence of considerable genetic polymorphism in them. Hence, this rapid and easy method could suitably be employed in carrying out significant molecular epidemiological studies and also in the large-scale epidemiological typing of this parasite. PMID- 11936501 TI - Survival of Cryptosporidium parvum oocysts recovered from experimentally contaminated oysters (Ostrea edulis) and clams (Tapes decussatus). AB - Samples of two species of shellfish that form part of the human food chain (the oyster Ostrea edulis and the marine clam Tapes decussatus) were experimentally contaminated with Cryptosporidium parvum oocysts. Changes in the viability of oocysts subsequently recovered from the shellfish were evaluated by means of an immunofluorescent antibody technique (IFAT) and inclusion/exclusion of the fluorogenic vital dye propidium iodide. There was a sharp decrease in oocyst viability during the first 4 days, with 15-25% viable oocysts remaining thereafter. In addition the infectivity of these oocysts at 10 and 31 days post contamination was demonstrated using a suckling murine model. PMID- 11936502 TI - Ultrastructural localization of glycoconjugates in Tritrichomonas foetus. AB - We have probed Tritrichomonas foetus, a protozoan parasite of the urogenital tract of cattle, with gold-labeled and fluorescent lectins, for the localization of glycoconjugates, at the cell surface and in internal cell compartments. The following lectins were used: Con A, PNA, WGA, BSI, BSII, HPA, WFA, SBA, LFA, HPA, and LCA. Carbohydrates were also localized using the Thiery's technique. Carbohydrate residues were observed in the Golgi and vesicles with all lectins used. However, the labeling pattern varied among the cisternae. A high heterogeneity of cell labeling was detected in the same preparation. The nucleus and the nuclear envelope were labeled with Con A, WGA, BSI, and UEA I, suggesting the presence of D-Man, and/or D-glucose, L-fucose, and GlcNAc. Lysosomes were labeled with Con A and intensely labeled with HPA. The intensity of labeling of the plasma and flagellar membranes varied according to the lectin used. PMID- 11936503 TI - Recombinant feline herpesvirus type 1 expressing Toxoplasma gondi ROP2 antigen inducible protective immunity in cats. AB - In order to investigate whether a recombinant viral vaccine composed of a feline herpesvirus type 1 (FHVI) vector and an immunogenic antigen of Toxoplasma gondii can induce a protective immunity in cats, a recombinant FHV1 expressing ROP2 antigen of T. gondii was prepared. We introduced a DNA fragment encoding 1-538 amino acid residues of ROP2 precursor into FHV1 genome under the control of a cytomegalovirus promoter. The recombinant FHV1 (FHV/ROP2) successfully expressed a 59 kDa antigen that was recognized by anti-ROP2 antibodies. Vaccination of cats with FHV/ROP2 induced serum IgG recognizing the native antigen. Moreover the antibodies inhibited the in vitro invasion of tachyzoites. The vaccination also accelerated the IgG response after T. gondii infection and reduced brain parasite load in cats. PMID- 11936504 TI - Taenia solium cysticercosis: lymphocytes in the inflammatory reaction in naturally infected pigs. AB - Immunohistochemistry was used to examine the type of lymphocytes in muscle taeniosis-cysticercosis in naturally infected pigs. The inflammatory response studied was classified into lesions of grades 1, 3, and 5. In grade 1, with a minimal inflammatory infiltrate consisting of eosinophils and a few mononuclear cells, the immunostaining showed more CD4+ cells than CD8+ cells and IgM cells. In grade 3, when the granulomatous reaction was not yet well developed and the destruction of the parasite began, CD4+ and Ig M+ were the predominant cells, although CD8+ cells showed a notable increase. In grade 5, with a few parasitic structures surrounded by an extensive granulomatous infiltrate, lymphocyte subsets were decreased in number and did not show differences from grade 1 except for IgM+ cells, which remained increased. The organization of an active inflammatory response against the metacestode of Taenia solium in pigs includes the sequential participation of CD4+, CD8+ and IgM+ lymphocytes. PMID- 11936505 TI - Could ivermectin have a synergic effect with albendazole in hydatidosis therapy? AB - The efficacy of ivermectin (IVM) alone, albendazole (ABZ) alone and a combination of both (IVM + ABZ) against Echinococcus granulosus protoscolices was studied by means of in vitro incubation. The maximum protoscolicidal effect was detected when we used IVM+ABZ in combination. In this case, protoscolex viability dropped to 35% in comparison with 50% for IVM alone and 82.5% with ABZ alone 18 days post incubation. Only the protoscolices incubated with IVM + ABZ did not develop into cysts following their inoculation into mice. The changes in ultrastructure induced in the protoscolices after the different drug incubations are also described. The incubation of cysts with IVM and IVM + ABZ shows that IVM + ABZ in combination is more effective than IVM alone as only when we incubated the cysts with IVM + ABZ did they completely lose their infectivity to mice. This occurred after only 10 days post-incubation. Cellular alterations were also more marked with IVM+ABZ incubations with: (1) the presence of residual bodies, (2) numerous lipids droplets and (3) vacuoles in the cytoplasm of cytons. However, the truncated microtriches and the nuclei remained unaltered after 10 days. PMID- 11936506 TI - Electron microscopic study on the endogenous development of Eimeria mulardi, Chauve, Reynaud and Gounel, 1994: a coccidium from the mule duck. AB - An electron microscopic study of the endogenous development of Eimeria mulardi Chauve, Reynaud and Gounel, 1994 was carried out in mule ducks which are hybrids of the domestic duck (Anas platyrhynchos) and the muscovy duck (Cairina moschata). All of the endogenous stages were seen within the nucleus of the host cell. Merozoites arose from ectomerogony and three mutually similar merogonies were noted. The asexual stages were found in leukocyte-like cells in the lamina propria of the jejunum, ileum and caecum, while the gamonts developed in glandular epithelial cells in the same part of the intestine. PMID- 11936507 TI - Synergistic in vitro antimalarial activity of plant extracts used as traditional herbal remedies in Mali. AB - In Mali, where malaria is endemic, plants are extensively used for treating periodic fevers and malaria. According to the advice of traditional medicine, plants are often mixed during the preparation of febrifugal decoctions. In previous studies, we demonstrated the potent in vitro antimalarial activity of extracts isolated from four plants commonly used in traditional remedies: Mitragyna inermis (Willd.) O. Kuntze, Rubiaceae, Nauclea latifolia (Sm.), Rubiaceae, Guiera senegalensis (Gmel.), Combretaceae, and Feretia apodanthera (Del.), Rubiaceae. In the present work, we evaluate the potent in vitro synergistic antimalarial interaction between these extracts, using standard isobologram analysis. Then, we evaluate their cytotoxicity on human monocytes and their mutagenic activity on an in vitro system of two beta-carboline alkaloids isolated from Guiera senegalensis (harman and tetrahydroharman). Three combinations demonstrate a strong, synergistic, inhibitory effect on in vitro plasmodial development and are devoid of cytotoxicity towards human cells. These results justify their use in association in traditional medicine. Moreover, tetrahydroharman, isolated from G. senegalensis, presents interesting antimalarial activity, no cytotoxicity and is not genotoxic in the Salmonella Ames test with and without metabolic activation. PMID- 11936508 TI - An anti-contamination cocktail for the in vitro isolation and cultivation of parasitic protozoa. AB - Contamination by bacteria or fungi is a frequent problem with the in vitro isolation and propagation of protozoan parasites. We developed an antibiotic combination of broad anti-contaminant activity but minimal toxicity to protozoa. This anti-contamination cocktail consists of penicillin G (60 microgml(-1)), kanamycin (100 microgml(-1)), chloramphenicol (10 microgml(-1)) and flucytosine (50 microgml(-1)). It is well tolerated by African and South American trypanosomes, Leishmania spp., Giardia duodenalis, Entamoeba histolytica and Plasmodium falciparum, and it provides an effective means for the treatment of contaminated cultures. The anti-contamination cocktail meets numerous prophylactic applications in which sterile handling is impracticable or impossible. In particular, it facilitates the direct in vitro isolation of parasites from an infected host. PMID- 11936509 TI - Neospora caninum is an invalid species name: an evaluation of facts and statements. AB - An evaluation of both the formal requirements of the International Rules of Zoological Nomenclature and the scientific reasons for the description of new genera and species shows that the name Neospora caninum is a nomen nudum. The only characteristic criteria for discriminating between the previously described species Hammondia heydorni and the proposed new species N. caninum (i.e. the lack of a parasitophorous vacuole) has been shown to be wrong in many publications. Furthermore, absolutely no criteria were presented as to why a new genus (i.e. Neospora) should be established besides the already existing genera Hammondia, Toxoplasma and Isospora. In addition, recent transmission experiments show that an oocyst isolate (from the faeces of dogs) is morphologically indistinguishable from H. heydorni [synonymous with Isospora bigemina - small form, Isospora heydorni (Tadros and Laarman 1976) and H. heydorni (Tadros and Laarman 1976) Dubey 1977] and is almost identical with respect to molecular biological features with the NC-1 strain of N. caninum. PMID- 11936510 TI - Ultrastructural alterations induced by nifurtimox and another nitro derivative on epimastigotes of Trypanosoma cruzi. AB - We report the ultrastructural alterations induced on epimastigotes by nifurtimox and 5-nitro-2-thienyl-malononitrile (5NO2TM), a novel compound with anti Trypanosoma cruzi activity. Parasites treated with concentrations of nifurtimox lower than usually employed for this kind of study showed vacuolisation, alterations of the mitochondria, the nucleus and the ribosomes. 5NO2TM caused the same kind of damage, but to a greater degree. This result correlates with the fact that cultures treated with this compound experienced a greater loss of viability. PMID- 11936511 TI - Legionella spp. in acute exacerbations of chronic obstructive pulmonary disease: what is the evidence? PMID- 11936512 TI - Minimal clinically significant difference in health status: the thorny path of health status measures? PMID- 11936513 TI - Serological evidence of Legionella species infection in acute exacerbation of COPD. AB - A prospective study was conducted to identify and characterize hospitalizations for acute exacerbations of chronic obstructive pulmonary disease (AECOPD) with serological evidence of infection with Legionella spp. (Lsp). Two-hundred and forty hospital admissions for AECOPD of 213 patients were included in the study. Paired sera were obtained for each of the admissions and were tested for 41 different serogroups of Lsp, using microimmunofluorescence-serology. Only a significant change in immunoglobulin-G and/or immunoglobulin-M antibody titres was considered diagnostic. In 40 admissions (16.7%) there was serological evidence of infection with Lsp (LspH). Legionella pneumophila 1 was identified in nine admissions, L. pneumophila 3-15 in 19 and nonpneumophila in 22. In 26 LspH (65%) there was serological evidence of infection with at least one other respiratory pathogen. Compared to the 200 admissions without Lsp (NLspH), the LspH patients were younger (p<0.05) and more hypoxaemic (p<0.04). None of the cases in the LspH group had an abrupt onset of disease, compared to 58 (29.0%) in the NLspH group (p<0.0001). The incidence of myalgia/arthralgia was 55% for LspH compared to 37% for NLspH (p<0.03). To conclude, serological evidence of infection with Legionella spp. is common among patients hospitalized with acute exacerbations of chronic obstructive pulmonary disease. In most hospital admissions with serological evidence of infection with Legionella spp. an additional respiratory pathogen can be identified. Acute exacerbation develops gradually in these patients and is characterized clinically by more systemic manifestations than hospital admissions without serological evidence of infection with Legionella spp. The true interpretation and practical relevance of these findings should be determined in further studies. PMID- 11936514 TI - Interpreting thresholds for a clinically significant change in health status in asthma and COPD. AB - Health status (or Health-Related Quality of Life) measurement is an established method for assessing the overall efficacy of treatments for asthma and chronic obstructive pulmonary disease (COPD). Such measurements can indicate the potential clinical significance of a treatment's effect. This paper is concerned with methods of estimating the threshold of clinical significance for three widely used health status questionnaires for asthma and COPD: the Asthma Quality of Life Questionnaire, Chronic Respiratory Questionnaire and St George's Respiratory Questionnaire. It discusses the methodology used to obtain such estimates and shows that the estimates appear to be fairly reliable; ie. for a given questionnaire, similar estimates may be obtained in different studies. These empirically derived thresholds are all mean estimates with confidence intervals around them. The presence of these confidence intervals affects the way in which the thresholds may be used to draw inferences concerning the clinical relevance of clinical trial results. A new system of judging the magnitude of clinically significant results is proposed. Finally, an attempt is made to translate these thresholds into scenarios that illustrate what a clinically significant change with treatment may mean to an individual patient. PMID- 11936515 TI - Interpretation of quality of life scores from the St George's Respiratory Questionnaire. AB - The aim of the study was to obtain the general population norms for the St. George's Respiratory Questionnaire (SGRQ), a specific questionnaire for respiratory diseases. The IBERPOC project was a cross-sectional study of representative samples of the general population aged between 40-69 yrs. The study sample was composed of 862 individuals. All participants considered as "probable cases" of chronic obstructive pulmonary disease (COPD) (n=460) were eligible to complete the SGRQ and among the rest of the nonprobable COPD participants (n=3,571), 10 individuals from each defined age and sex group were eligible (n=402). Weights were applied to restore general population representativity of the sample. Mean forced expiratory volume in one second (FEV1) predicted was 89.4% (SD=16.5%; range: 16-131%). Chronbach's alpha coefficients were >0.7 in the symptoms, activity and impact scales, and >0.9 in the overall scale. Symptom scale score was significantly higher among males (11.6 versus 7.8; p<0.01) and activity scale score was significantly higher among females (12.2 versus 14.6; p=0.04). In a multiple linear regression model, respiratory diseases (asthma and COPD) and FEV1 % over pred showed the strongest association with the SGRQ total score. Smoking, sex, age and education were independently associated with the total SGRQ score. These results indicate that individuals from the general population presented some of the problems that are important when measuring health-related quality of life in respiratory patients, and provide St George's Respiratory Questionnaire norms, a useful method for interpreting the St George's Respiratory Questionnaire score in a given patient or study samples. PMID- 11936516 TI - A national study of medical care expenditures for respiratory conditions. AB - This study was undertaken to estimate the magnitude of medical care expenditures among persons with respiratory conditions in the USA in 1996, and the increment in expenditures attributable to these conditions. The study data were derived from the 1996 Medical Expenditure Panel Survey, a national sample of 21,571 persons. Of the 21,571, 1,027 reported one or more respiratory condition. After weighting, the individuals may represent about 12.1 million persons in the USA. All medical care expenditures of these individuals were tabulated, stratified by comorbidity status, and then compared to those among persons with nonrespiratory conditions or with no conditions. Regression techniques were then used to estimate the increment of healthcare expenditures attributable to the respiratory conditions. From a national total of $45.3 billion, medical care expenditures averaged $3,753 among persons with respiratory conditions. Hospital stays comprised the largest component (45%). The per capita increment in total expenditures attributable to respiratory conditions ranged from $1,003-2,588, from a national total ranging from $12.1-31.3 billion. The total medical care expenditure of persons with respiratory conditions was estimated to be $45.3 billion, of which $12.1-31.3 billion represents an increment in expenditures associated with the conditions themselves. PMID- 11936517 TI - Effects of lung volume reduction surgery in hamsters with elastase-induced emphysema. AB - Lung volume reduction surgery (LVRS) has been shown to improve respiratory mechanics in selected patients with severe emphysema. This is thought to be due to an improvement in lung elastic recoil. This study was aimed at gaining further understanding about the effects of LVRS on respiratory mechanics and airway function. Control hamsters instilled with saline (Ctrl; n=8) were compared with emphysematous animals that underwent either a sham operation (Sham; n=7) or an LVRS (LVRS; n=7). As expected, there was a significant increase in the static lung volumes in the Sham as compared to the Ctrl group and a significant decrease of these volumes in LVRS as compared to the Sham group. Surprisingly, emphysema was associated with a significant increase and LVRS with a significant decrease in vital capacity. Despite a tendency toward an increase in lung compliance as compared to Sham, indices of maximal expiratory flows tended to decrease with LVRS. As opposed to humans, there was no change in the distribution of airway diameters in Sham compared to Ctrl. These findings appear to be largely explained by the high compliance of the hamster chest wall. This allows for better matching between the emphysematous lung and the chest-wall sizes than in humans. PMID- 11936518 TI - Effect of chest wall vibration on the canine diaphragm during breathing. AB - High-frequency mechanical vibration of the ribcage reduces dyspnoea in patients with chronic obstructive pulmonary disease, and the suggestion has been made that this effect might be related to a decrease in central respiratory drive resulting from an increase in afferent inputs from intercostal muscles. In the present studies, the effects of ribcage vibration on central respiratory drive have been assessed without the confounding influence of conscious reactions. The electromyographic (EMG) activity of the diaphragm and the changes in pleural (Ppl) and abdominal (Pab) pressure were measured in six anaesthetized, spontaneously-breathing dogs while the rostral, the middle, or the caudal portion of the ribcage was vibrated at intervals during inspiration. The EMG activity of the external and parasternal intercostals was also measured. Ribcage vibration consistently elicited a marked increase in the inspiratory EMG activity recorded from the external intercostals, thus indicating that the procedure did activate intercostal muscle spindles. However, no alteration in diaphragmatic or parasternal intercostal EMG activity was seen in any animal. Transdiaphragmatic pressure and the relationship between deltaPab and deltaPpl during inspiration were also unaltered. The authors conclude that ribcage vibration and, with it, stimulation of external intercostal muscle spindles has no significant influence on phrenic motoneurones or on medullary inspiratory neurones. It is unlikely, therefore, that the beneficial effect of the procedure on dyspnoea results from a specific reduction in central respiratory drive. PMID- 11936519 TI - Relationship between supramaximal flow during cough and mortality in motor neurone disease. AB - The main function of cough is clearance of intrathoracic airways. A normal cough is characterized by a transient increase in expiratory flow above the maximal flow-volume loop envelope, known as cough "spikes". They may be absent in patients with motor neurone disease. The relationship between cough pattern, pulmonary function and survival was studied. Fifty-three patients were recruited (25 bulbar). Vital capacity, maximal inspiratory and expiratory mouth pressures and cough flow/volume curves were performed on all patients, and the presence or absence of spikes were recorded. The primary endpoints were mortality or initiation of ventilatory support over a period of 18 months. Thirty-five patients died over the 18-month period of the study (including the six who were started on noninvasive ventilation). Twelve of the 24 patients with spikes died compared to 23 out of 29 patients without spikes (p<0.05). Patients without spikes were more likely to be bulbar on clinical grounds (p<0.0001) and had poorer lung function. The results showed an association between the absence of cough spikes and increased mortality. However the main determinants of survival in motor neurone disease are age, vital capacity and inspiratory mouth pressure, and it remains to be shown whether regular monitoring of cough conveys any additional advantage. PMID- 11936520 TI - Serum surfactant proteins-A and -D as biomarkers in idiopathic pulmonary fibrosis. AB - Idiopathic pulmonary fibrosis (IPF) has a high mortality rate, and current therapies are only marginally effective. A serum biomarker that predicts clinical outcome would be useful to stage disease, indicate prognosis and the need for aggressive therapy, and help stratify patients for clinical trials. The goals of this study were to determine whether serum levels of surfactant protein-A (SP-A) or surfactant protein-D (SP-D) would distinguish between IPF and other types of interstitial lung disease and whether serum SP-A or SP-D levels predict outcome in patients with IPF. The authors found that serum SP-A and SP-D levels were significantly elevated in patients with IPF and systemic sclerosis compared to sarcoidosis, beryllium disease and normal controls, and that SP-D correlated with radiographic abnormalities in patients with IPF. In addition, the authors found that both serum SP-A and SP-D levels were highly predictive of survival in patients with IPF. This is the largest North American data set of surfactant protein measurements in idiopathic pulmonary fibrosis and the first report using multivariate analysis comparing serum surfactant proteins-A and -D to other commonly measured predictors of survival in idiopathic pulmonary fibrosis. Based on these results, the authors propose that serum surfactant proteins may prove to be useful biomarkers in patients with idiopathic pulmonary fibrosis. PMID- 11936521 TI - Effects of inducible nitric oxide synthase and xanthine oxidase inhibitors on SEB induced interstitial pneumonia in mice. AB - The authors have previously reported that intratracheal instillation of staphylococcal enterotoxin-B (SEB) induced interstitial pneumonia (IP) in autoimmune-prone mice. SEB-reactive T-cells were critically involved in the development of IP in this model. Concern has arisen about the hazards of reactive oxygen species (ROS) and reactive nitrogen species (RNS) in the process of lung injury and fibrosis. Therefore, the involvement of nitric oxide (NO) and superoxide anion (O2-) in the pathogenesis of IP in this autoimmune-prone model has been investigated. Nitrite/nitrate levels were increased in bronchoalveolar lavage (BAL) fluid and serum from SEB-injected mice. The signal of the NO-(N (dithiocarboxy) sarcosine)2-Fe2+ complex was detected in the SEB-injected lung and whole blood by electron paramagnetic resonance (EPR) spectroscopy. NO production was significantly decreased by aminoguanidine (AG) treatment. Xanthine oxidase (XO) activity in the lung, BAL fluid, and plasma was increased with instillation of SEB, and 4-amino-6-hydroxypyrazolo(3,4-d)-pyrimidine (AHPP) significantly inhibited XO activity. Moreover, both AG and AHPP significantly decreased production of pro-inflammatory cytokines, numbers of infiltrated cells in BAL fluid, and the area of thickened alveolar septa in the SEB-injected lung. In conclusion, the overproduction of nitric oxide and super oxide anion were implicated in the pathogenesis of interstitial pneumonia, and inducible nitric oxide synthase and xanthine oxidase inhibitors had protective effects against interstitial pneumonia in this model. PMID- 11936522 TI - Survey of flexible fibreoptic bronchoscopy in the United Kingdom. AB - The practice of flexible bronchoscopy is not standardized. Current guidelines are concerned primarily with safety aspects of the procedure. In view of this, and the authors' own observations of individual variation in preparation and technique, a national survey of bronchoscopic procedure was performed to assess physicians' methods. A structured questionnaire was mailed to 547 consultant physicians in adult respiratory medicine. Physicians' routines of patient preparation, drug therapy, sampling methods, and experience of complications with the flexible bronchoscope were assessed. A 60% response (328 physicians) was obtained. Patient consent was obtained by a junior doctor in 31% of replies. 205 (63%) physicians gave benzodiazepine sedation, 46 (14%) used opioid, and 38 (12%) administered both. Ninety-four (29%) physicians prescribed an antimuscarinic agent, and 235 (74%) gave antibiotics to patients with mechanical heart valves. Only 22% of physicians monitored electrocardiogram and 10% monitored blood pressure during all procedures. Marked variance was noted in sampling routines of suspected lung tumours. Physicians who used fluoroscopic guidance for transbronchial lung biopsy reported a significantly lower incidence of pneumothorax requiring drain insertion over the previous 12 months compared to those who did not (2.68 of 1000 versus 9.17 of 1000, (p<0.03)), but no difference in the total incidence of pneumothorax. Only 87 (27%) of responders had performed transbronchial needle aspiration sampling over the previous 12 months. The preparation and practice of flexible bronchoscopy varies greatly for each physician. Use of radiographical screening for performing transbronchial lung biopsy was associated with a lower likelihood of pneumothorax requiring chest tube drainage. PMID- 11936524 TI - Achilles tendon disease in lung transplant recipients: association with ciprofloxacin. AB - Achilles tendonitis or rupture are uncommon complications following the use of fluoroquinolones, with a reported incidence in the general population of 0.4%. The aims of the current study were to determine the incidence of Achilles tendon disease (ATD) in lung transplant recipients (LTR) and to identify risk factors. Questionnaires were sent to 150 LTR of whom 101 responded (67%). Twenty-two LTR (21.8%) experienced ATD (tendonitis 16, rupture six). The mean age of LTR who developed ATD was 52.9+/-6.1 yrs (range: 19-63.5 yrs). Only the use of ciprofloxacin was significantly associated with ATD (p<0.05). Age, sex, underlying disease necessitating transplantation, serum creatinine and cyclosporine levels were not associated with ATD. The association between ciprofloxacin and ATD was not dose related. Of the 72 LTR who had received ciprofloxacin, 20 (28%) developed ATD (tendonitis 15, rupture five). In patients receiving ciprofloxacin, there was no association between the mean cumulative dose of prednisolone and ATD. Tendon rupture occurred with a lower ciprofloxacin dosage than tendonitis and the mean recovery duration was significantly longer. To conclude, lung transplant recipients receiving ciprofloxacin are at significant risk of developing Achilles tendon disease. The association between ciprofloxacin and Achilles tendon disease appears to be idiosyncratic rather than dose-related. PMID- 11936523 TI - Regular follow-up after curative resection of nonsmall cell lung cancer: a real benefit for patients? AB - Even though complete resection is regarded as the only curative treatment for nonsmall cell lung cancer (NSCLC), >50% of resected patients die from a recurrence or a second primary tumour of the lung within 5 yrs. It remains unclear, whether follow-up in these patients is cost-effective and whether it can improve the outcome due to early detection of recurrent tumour. The benefit of regular follow-up in a consecutive series of 563 patients, who had undergone potentially curative resection for NSCLC at the University Hospital, was analysed. The follow-up consisted of clinical visits and chest radiography according to a standard protocol for up to 10 yrs. Survival rates were estimated using the Kaplan-Meier analysis method and the cost-effectiveness of the follow up programme was assessed. A total of 23 patients (6.4% of the group with lobectomy) underwent further operation with curative intent for a second pulmonary malignancy. The regular follow-up over a 10-yr period provided the chance for a second curative treatment to 3.8% of all patients. The calculated costs per life-yr gained were 90,000 Swiss Francs. The cost-effectiveness of the follow-up protocol was far above those of comparable large-scale surveillance programmes. Based on these data, the intensity and duration of the follow-up was reduced. PMID- 11936525 TI - Increased soluble CD14 in bronchoalveolar lavage fluid of stable lung transplant recipients. AB - Macrophages, neutrophils and infection have been implicated in the genesis of the bronchiolitis obliterans syndrome (BOS) post lung transplantation. sCD14 is a soluble form of a shed-cell surface protein. It is capable of promoting cytokine induced inflammation and it's presence in clinically stable lung transplant recipients (LTR) might be important as an early marker of BOS. Bronchalveolar lavage (BAL) and blood samples were taken from 26 stable LTR, at or near their best forced expiratory volume in one second who were free from infection. sCD14 levels were measured via enzyme-linked immunosorbent assay. Cell counts were performed on unfiltered BAL. LTR neutrophil count, BAL sCD14 and serum sCD14 levels were higher than controls (median 3.8% versus 1.3%, p<0.05; 11.5 ng x mL( 1) versus 6 ng x mL(-1), p<0.001; 6.2 microg x mL(-1) versus 2.4 microg x mL(-1), p<0.05, respectively). BAL albumin and sCD14 correlated (regression coefficient: 0.77, p<0.001). In this hypothesis-generating, cross-sectional study, the authors have described for the first time soluble CD14 levels in the bronchoalveolar lavage and serum of stable lung transplant recipients, and show that these are elevated compared to controls. This is a practicable candidate marker system, which can be tested for a predictive role in bronchiolitis obliterans syndrome following lung transplantation. The origin of this cellular protein and its temporal relationship to the development of the bronchiolitis obliterans syndrome remains to be elucidated in more definitive longitudinal studies, which should include other measurements potentially relevant to the innate immune system, such as bronchoalveolar lavage endotoxin levels. PMID- 11936526 TI - Associations between markers of respiratory morbidity in European children. AB - School-aged children often experience acute respiratory symptoms. In a multicentre European study, the association between chronic respiratory symptoms (reported in a questionnaire), skin-prick test results, and lung function, and the occurrence of acute respiratory morbidity, was examined among children aged 6 12 yrs with chronic respiratory symptoms. Children with chronic respiratory symptoms, living in 10 European countries, were selected from a parent-completed questionnaire (n=4,307). Atopy was measured with skin-prick tests, and lung function with spirometry. A total of 1,854 (86% of those in the initial cohort) children kept a successful daily diary regarding their respiratory symptoms for 2 3 months. In multivariate logistic regression analyses, children with asthmatic symptoms, particularly those with doctor-diagnosed asthma, had a greater risk of occurrence of lower respiratory symptoms (odds ratio (OR): 6.12; 95% confidence interval (CI): 4.99-8.35) than children with a dry nocturnal cough as their only symptom. Atopy, particularly a positive reaction to indoor allergens, was significantly associated with occurrence of lower respiratory symptoms. For atopy the OR was 1.62 (95% CI: 1.34-1.96). A reduced level of maximal mid-expiratory flow was associated with an increased risk of lower respiratory symptoms, cough and phlegm. The associations were similar in Scandinavia, Central Eastern, Western and Southern Europe. To conclude, asthmatic symptoms reported in a questionnaire, atopic status and a reduced level of maximal mid-expiratory flow were associated with the occurrence of acute respiratory symptoms, especially those of lower respiratory symptoms. PMID- 11936527 TI - Elevated nitrite in breath condensates of children with respiratory disease. AB - The aim of the study was to determine the differences in nitrite, in the exhaled breath condensates of healthy children and those children with asthma, cystic fibrosis (CF) and nonasthmatic, episodic cough. Breath condensates were obtained from 66 children (43 males:23 females, 3.1-16 yrs) and included 29 asthmatics, 12 clinically stable CF patients, 12 children with cough but not asthma and 13 healthy volunteers. The collected condensate was assayed colourimetrically using the Griess reaction to determine nitrite concentrations. Patients with CF (median: 5-95% percentiles; 2.02: 0.43-6.37 microM) or asthma (2.10: 0.63-5.45 microM) had significantly higher levels of nitrite compared to healthy subjects (0.41: 0.13-1.83 microM; p<0.05) or subjects with cough (0.75: 0.03-1.75 microM; p<0.05). Airway inflammation, as assessed by the nitrite in breath condensates, is present in children with asthma and cystic fibrosis, but not those children with nonasthmatic, episodic cough. Nitrite can be conveniently, cheaply and rapidly measured in breath condensates of children as young as 3 yrs of age, and may prove useful for the assessment of airway inflammation in children with respiratory disease. PMID- 11936528 TI - Tidal expiratory flow limitation and chronic dyspnoea in patients with cystic fibrosis. AB - Cystic fibrosis (CF) eventually leads to hyperinflation linked to tidal expiratory flow limitation (FL) and ventilatory failure. Presence of FL was assessed at rest in 22 seated children and adults with CF (forced expiratory volume in one second (FEV1) range: 16-92% predicted), using both the negative expiratory pressure (NEP) technique and the "conventional" method based on comparison of tidal and maximal expiratory flow/volume curves. In addition, chronic dyspnoea was scored with the modified Medical Research Council (MRC) scale. Measurements were made before and 15 min after inhalation of salbutamol. With NEP, FL was present in only three malnourished patients, who had the lowest FEV1 values (16-27% pred) and claimed very severe dyspnoea (MRC score 5). By contrast, an additional seven patients were classified as FL with the conventional method. Six of these patients had little or no dyspnoea (MRC scores 0-1). Salbutamol administration had no effect on the extent of FL, and the concomitant decrease in functional residual capacity (FRC) was too small to play any clinically significant role. This study concluded that in seated patients with cystic fibrosis, expiratory flow limitation is absent at rest, unless the forced expiratory volume in one second is <30% predicted. If present, expiratory flow limitation is associated with severe chronic dyspnoea. The conventional method for assessing expiratory flow limitation is not reliable and bronchodilator administration has little effect on expiratory flow limitation. PMID- 11936529 TI - Health-related quality of life in obstructive sleep apnoea. AB - The identification of the areas of quality of life (QoL) most likely to be affected by obstructive sleep apnoea (OSA) would be an important step in the evaluation of the impact of the disease and its treatment modalities. The objective of this study was to describe the impact of OSA on patients' QoL. A list of 186 items potentially related to QoL of patients with OSA was constructed. From this list, consecutive patients were asked, at the time of the diagnosis, to identify the most significant items and to grade their importance on a 5-point scale. The item impact was determined from the proportion of patients who identified it as important, and the mean importance score attributed to this item (impact score=frequency x importance). One hundred patients (82 male; mean age: 51 yrs) were interviewed. The items having the most important impact on QoL clustered into five domains: 1) daytime symptoms; 2) nocturnal symptoms; 3) limitation of activities; 4) emotions; and 5) interpersonal relationships. The impact of obstructive sleep apnoea on quality of life is not limited to excessive daytime sleepiness. Obstructive sleep apnoea significantly contributes to the impairment of all domains of what is usually referred to as "health-related quality of life". PMID- 11936530 TI - Sleep quality and daytime function in adults with cystic fibrosis and severe lung disease. AB - It was hypothesized that adult cystic fibrosis (CF) patients with severe lung disease have impaired daytime function related to nocturnal hypoxaemia and sleep disruption. Nineteen CF patients (forced expiratory volume in one second 28+/-7% predicted) and 10 healthy subjects completed sleep diaries, overnight polysomnography (PSG), and assessment of daytime sleepiness and neurocognitive function. CF patients tended to report more awakenings (0.7+/-0.5 versus 0.3+/ 0.2 x h(-1), p=0.08), and PSG revealed reduced sleep efficiency (71+/-25 versus 93+/-4%, p=0.004) and a higher frequency of awakenings (4.2+/-2.7 versus 2.4+/ 1.4 x h(-1), p=0.06). Mean arterial oxygen saturation during sleep was lower in CF patients (84.4+/-6.8 versus 94.3+/-1.5%, p<0.0001) and was associated with reduced sleep efficiency (regression coefficient (r)=0.57, p=0.014). CF patients had short sleep latency on the multiple sleep latency test (6.7+/-3 min). The CF group reported lower levels of activation and happiness and greater levels of fatigue (p<0.01), which correlated with indices of sleep loss, such as sleep efficiency (r=0.47, p=10.05). Objective neurocognitive performance was also impaired in CF patients, reflected by lower throughput for simple addition/subtraction, serial reaction and colour-word conflict. The authors concluded that adult cystic fibrosis patients with severe lung disease have impaired neurocognitive function and daytime sleepiness, which is partly related to chronic sleep loss and nocturnal hypoxaemia. PMID- 11936531 TI - Obstructive sleep apnoea and urine catecholamines in hypertensive males: a population-based study. AB - Studies addressing the relationship between obstructive sleep apnoea (OSA) and sympathoadrenal activity have been criticized for poor control of factors known to confound sympathetic function, including hypertension. The aim of this study was to investigate the relationship between OSA and urinary catecholamines in a population-based sample of hypertensive males. In 1994, 2,668 males aged 40-79 yrs answered a questionnaire regarding sleep disorders and somatic diseases. Of those who reported hypertension, an age-stratified sample of 116 was selected for monitoring of breathing during sleep and overnight urine analysis. Subjects with OSA, defined as apnoea-hypopnoea index > or = 10 x h(-1), had higher concentrations of urinary normetanephrine (182+/-57 versus 141+/-45 micromol x mol(-1) creatinine, p<0.001) and metanephrine (70+/-28 versus 61+/-28 micromol mol(-1) creatinine, p<0.05) in comparison to subjects without OSA. In a multiple regression analysis, there was an association between variables of sleep disordered breathing and normetanephrine and metanephrine concentrations, independent of major confounding factors. The authors concluded that, in a population-based sample of hypertensive males, obstructive sleep apnoea is associated with increased urinary concentrations of extraneuronal metabolites of catecholamines independent of major confounding factors, suggesting increased sympathoadrenal activity. Elevated sympathoadrenal activity may explain the increased cardiovascular morbidity associated with obstructive sleep apnoea. PMID- 11936532 TI - Aerosolized iloprost induces a mild but sustained inhibition of platelet aggregation. AB - Pathological studies have revealed that one of the main features encountered in the pulmonary vasculature of patients with pulmonary hypertension is the presence of thrombotic lesions. Open pilot studies have indicated that aerosolized iloprost may have beneficial effects in patients with pulmonary hypertension. The effects of aerosolized iloprost on platelet function and plasma cyclic adenosine monophosphate (cAMP) were studied. Platelet aggregation and plasma cAMP were measured at baseline, 30 min, 4 and 6 h after inhalation of 15 microg iloprost in 10 healthy volunteers. Maximal platelet aggregation in response to adenosine diphosphate (ADP) (2 and 6 micromol x L(-1)), collagen (2.5 and 5 microg x mL( 1)), epinephrine (1.25 and 5 micromol x L(-1)) and arachidonic acid (0.5 mg x mL( 1)) was measured. Platelet aggregation was significantly inhibited at 30 min in response to ADP (2 and 6 micromol x L(-1), epinephrine (1.25 and 5 micromol x L( 1)) and collagen (2.5 microg x mL(-1)). It was still inhibited at 4 h in response to the same agents, but normalized at 6 h. cAMP increased at 30 min, from 27.3+/ 1.2 to 31.8+/-1.2 nmol x L(-1), remained increased at 4 h (29.2+/-1.3 nmol x L( 1)) and normalized at 6 h (27.4+/-1.1 nmol x L(-1)). Aerosolized iloprost induced a mild but sustained inhibition of platelet aggregation. Platelet aggregation inhibition may be one of the mechanisms which explains the beneficial effect of repeated inhalation of iloprost in pulmonary hypertension. PMID- 11936533 TI - Role of tyrosine phosphatase in the modulation of pulmonary vascular tone. AB - In the vascular system, synthesis of the potent vasodilator nitric oxide (NO) is tightly regulated by the constitutively expressed endothelial NO synthase (eNOS). Activity of eNOS is controlled by Ca2+/calmodulin and various seryl/threonyl protein kinases. Less is known about the importance of phosphorylation and dephosphorylation of tyrosyl residues. Therefore the role of tyrosine phosphatase on the modulation of isolated rat pulmonary artery tone has been assessed. Inhibition of tyrosine phosphatase by sodium orthovanadate (SOV, 1x10(-6) M) significantly: 1) increased phenylephrine-induced vasoconstriction and 2) decreased endothelium-dependent relaxation to acetylcholine, but had no effect on endothelium-independent relaxation to the NO donor, sodium nitroprusside. In phenylephrine-precontracted pulmonary arterial rings, SOV (1x10(-7)-1x10(-5) M) had no effect on vascular tone but significantly relaxed rings which were pretreated with the NO-synthase inhibitor, N(omega)-nitro-L-arginine-methyl ester (L-NAME). SOV-induced relaxation in the presence of L-NAME was, however, abolished by glibenclamide. In conclusion, inhibition of tyrosine phosphatase altered pulmonary vascular tone by increasing vasoconstrictor response to phenylephrine and decreasing endothelium-dependent relaxation to acetylcholine. Furthermore, the tyrosine phosphatase inhibitor, sodium orthovanadate, exhibited original vasodilator properties which were only observed when nitric oxide synthesis was inhibited. Thus a new pathway involving the inhibitory effect of nitric oxide on a glibenclamide-sensitive diffusible relaxing factor, that might play an important role in the control of pulmonary vascular tone is described. PMID- 11936534 TI - Recording flow in the first second of a maximal forced expiratory manoeuvre: influence of frequency content. AB - The frequency content of the first second of the maximum forced expiratory manoeuvre (MFEM) was measured to determine if the currently accepted frequency limit of 20 Hz for MFEM is adequate for recording peak expiratory flow (PEF). The frequency response of a Fleisch pneumotachograph (PT) was measured and used to record MFEM from 24 patients attending a lung-function laboratory and 26 normal volunteers. The first 1.024 s of the signal recorded at 1,000 Hz for that blow with maximum PEF, underwent fast Fourier transformation using a triangular window function, applied after 0.75 s to reduce flow linearly to zero. All the frequencies above a set limit were removed, followed by inverse transformation to reconstitute the blow. The limits for this frequency cut-off were progressively varied from 100 Hz down to 15 Hz, with the resulting PEF being compared with the PEF from the reconstituted blow with no frequency reduction. The average+/-SD age for the group was 47+/-18 yrs and the average PEF was 450+/-187 L x min(-1), which, when expressed as a standardized residual, was 0.1+/-2.1, with a range from -4.5-3.9 indicating a good spread around normal values. Average rise time to PEF was 83+/-38 ms and dwell time >90% PEF was 45+/-25 ms. Cut-off >20 Hz reduced the mean PEF of the group by 8.5 L x min(-1) (95% confidence limit 5.5-11.4 L x min(-1)), whereas cut-off >30 Hz reduced mean PEF by 4.4 L x min(-1) (2.6-6.2). In the present study subjects, 30 Hz was on the 95th percentile of frequencies for defining the upper limit for 98% of the power spectrum for the first second of the blow. It has been shown that there are frequencies >20 Hz that contribute to peak expiratory flow enough to influence readings made using conventional hand held peak expiratory flow meters, such as the mini-Wright. Devices used for recording flow from a maximum forced expiratory manoeuvre should therefore have an adequate frequency response of up to 30 Hz. PMID- 11936535 TI - Free, soluble interleukin-17 protein during severe inflammation in human airways. AB - Studies in rodents indicate that the cytokine, interleukin (IL)-17, links the activation of T-lymphocytes to neutrophilic inflammation. The aim of the current study was to determine whether free, soluble IL-17 protein can be released during severe inflammation in human airways. Fifteen healthy subjects were exposed to a swine confinement in order to induce severe inflammation characterized by high neutrophil numbers in the airways. Bronchoalveolar lavage (BAL) fluid was harvested 2 weeks prior to and 24 h after this exposure and the concentration of IL-17 protein was measured using an enzyme-linked immunosorbent assay. Total and cell differential counts were also performed in BAL fluid. Prior to exposure to the swine confinement, the concentration of IL-17 in BAL fluid was low (<7.8 pg x mL(-1)) in 14 out of 15 subjects. However, exposure to the swine confinement caused an increase in IL-17 in 13 out of 15 subjects (median IL-17 concentration of 26.9 pg x mL(-1)). This exposure also caused a 51-fold increase in the concentration of neutrophils in BAL fluid. To conclude, free, soluble interleukin 17 protein can be released during severe inflammation characterized by high neutrophil numbers in human airways. The significance of interleukin-17 in inflammatory airway diseases therefore deserves further evaluation. PMID- 11936537 TI - Role of persistent infection in the control and severity of asthma: focus on Chlamydia pneumoniae. AB - Conventional risk factors have been unable to explain most of the substantial increase in the prevalence of asthma observed in many countries during the last few decades. Much attention has been directed at the "hygiene hypothesis", the apparent inverse relationship between intense systemic childhood infections and the subsequent development of asthma and atopy. However, it is not only the absence or scarcity of infections, but the prolonged presence of certain microorganisms in the lungs that may be involved in the development of asthma. Accumulating evidence suggests that Chlamydia pneumoniae, an intracellular ubiquitous pathogen with an innate propensity to persist and cause chronic infections, may be associated with asthma. This microorganism can achieve a state of "latency" in which it is viable but dormant and does not multiply. During this state, however, chlamydia continues to synthesize the "stress" protein, a 60-kDa heat shock protein (hsp60). This protein is able to elicit a strong host inflammatory response at sites of its production and appears to be involved in tissue injury and scarring processes. As inflammation has been found to be present in almost all asthmatics, whatever the severity and aetiology of the disease, inhaled glucocorticoids now have an established position in the treatment of early stages. However, corticosteroids negatively affect many aspects of cell-mediated immunity and favour the shift from a T-helper-1-type response towards a T-helper-2-type response. Corticosteroids may thus severely deteriorate the host's ability to eradicate an intracellular pathogen, such as Chlamydia pneumoniae, which requires a properly functioning cell-mediated (T helper-1-type) immune response to be cleared. These drugs are also able to reactivate persistent Chlamydia to an active growth phase, which, by increasing the production of pro-inflammatory cytokines at the site of infection, can further amplify inflammation in the airways of patients with asthma. PMID- 11936536 TI - Interactions between respiratory tract infections and atopy in the aetiology of asthma. AB - The prevalence of asthma, in particular atopic asthma, has markedly increased in recent years. Accumulating evidence suggests that environmental factors associated with allergic sensitization and exposure to microbial stimuli during infancy and early childhood, are associated with these changes in prevalence. However, considerable controversy surrounds the role of microbial agents, as evidence has been presented for both positive and negative effects in this context. The review below focuses upon interactions between immune competence during infancy, the development of T-helper (Th)1-polarized versus Th2-polarized memory against inhalant allergens, and susceptibility to virus infection. In particular, recent finding are highlighted which suggest that delayed postnatal maturation of Th1 function is associated with increased risk for early postnatal sensitization to inhalant allergens, and also with risk for viral bronchiolitis during infancy. Variations in the kinetics of postnatal maturation of T-helper 1 function may in part be attributable to polymorphisms in the CD14 gene, which influence host responsiveness both to bacterial as well as viral stimuli. PMID- 11936538 TI - Anti-angiogenic strategies and vascular targeting in the treatment of lung cancer. AB - The generation of new blood vessels, angiogenesis, is important for tumour proliferation and metastasis. This involves a number of interacting processes and factors, such as growth factors and the receptor tyrosine kinases, matrix metalloproteinases and integrins. Studies have shown that tumour vascularity and the overexpression of growth factors and their receptors are of independent prognostic importance in different cancers, including lung cancer. The present article provides a background to angiogenesis and describes the potential targets for anti-angiogenic and vascular targeting strategies in cancer, focusing specifically on carcinoma of the lung. It also describes the anti-angiogenic drugs presently under phase I, II and III investigation and highlights some of the problems associated with the standard methodologies for assessing tumour response and drug efficacy using these agents. PMID- 11936539 TI - Pulmonary hypertension in collagen vascular disease. AB - Pulmonary hypertension is a serious but often overlooked complication in collagen vascular disease. The understanding of the development of pulmonary hypertension has increased substantially during the last years. Abnormal proliferation of pulmonary vascular cells is now being regarded as a predominant process leading to pulmonary vascular obliteration. Medical therapy focuses on prostacyclin treatment, which has been shown to improve exercise capacity and haemodynamic variables in patients with several collagen vascular diseases and pulmonary arterial hypertension. Continuous intravenous prostacyclin remains the standard treatment of associated pulmonary hypertension but less invasive alternatives such as subcutaneous treprostinil, oral beraprost or aerosolized iloprost, as well as, novel substances such as endothelin receptor antagonists may be appropriate for selected patients. PMID- 11936540 TI - Statin-induced fibrotic nonspecific interstitial pneumonia. AB - Statins inhibit the 3-hydroxy-3-methylglutaryl coenzyme A reductase, reduce the serum level of low-density lipoprotein cholesterol, and are extensively prescribed to prevent cardiovascular mortality and morbidity. Few systemic adverse effects, such as pseudopolymyositis, lupus-like syndromes, and anecdotal hypersensitivity pneumonitis, have been reported. A simvastatin-induced diffuse interstitial pneumonia associated with a nonspecific interstitial pneumonia pattern at histological analysis is repoted here. Ultrastructural analysis showed a diffuse cytoplasmic accumulation of intralysosomial lamellar inclusions in type II pneumonocytes, histiocytes and endothelial cells, suggesting a shared pathogenesis with amphiphilic drug-induced toxic lung injury. Because statins are increasingly prescribed, statin-induced interstitial lung disorders may be more frequently observed and early recognition will be required. PMID- 11936541 TI - An 11-yr-old male with pneumonia and persistent airway obstruction. PMID- 11936542 TI - Effects of posting self-set goals on collegiate football players' skill execution during practice and games. AB - The effects of self-set goals and public posting on athletic performance of 5 collegiate football players was studied. All players were linebackers on a National Association of Intercollegiate Athletics Division II football team. The dependent variables were the percentage of correct occasions when the linebacker (a) positioned himself to cover a specified area on the field during a pass or from the line of scrimmage during a run; (b) moved to the correct position in response to the positioning of the offense; and (c) tackled and stopped the progress of the ball carrier. A multiple baseline design across behaviors showed an immediate increase in the practice performance of the players and a corresponding increase in game performance following introduction of the independent variable. This study extends research using public posting in sport by demonstrating the effects of player-determined goals and public posting of goal attainment. PMID- 11936543 TI - An application of the matching law to severe problem behavior. AB - We evaluated problem behavior and appropriate behavior using the matching equation with 4 individuals with developmental disabilities. Descriptive observations were conducted during interactions between the participants and their primary care providers in either a clinical laboratory environment (3 participants) or the participant's home (1 participant). Data were recorded on potential reinforcers, problem behavior, and appropriate behavior. After identifying the reinforcers that maintained each participant's problem behavior by way of functional analysis, the descriptive data were analyzed retrospectively, based on the matching equation. Results showed that the proportional rate of problem behavior relative to appropriate behavior approximately matched the proportional rate of reinforcement for problem behavior for all participants. The results extend prior research because a functional analysis was conducted and because multiple sources of reinforcement (other than attention) were evaluated. Methodological constraints were identified, which may limit the application of the matching law on both practical and conceptual levels. PMID- 11936544 TI - Reinforcement magnitude and responding during treatment with differential reinforcement. AB - Basic findings indicate that the amount or magnitude of reinforcement can influence free-operant responding prior to and during extinction. In this study, the relation between reinforcement magnitude and adaptive behavior was evaluated with 3 children as part of treatment with differential reinforcement. In the first experiment, a communicative response was shaped and maintained by the same reinforcer that was found to maintain problem behavior. Two reinforcement magnitudes (20-s or 60-s access to toys or escape from demands) were compared and found to be associated with similar levels of resistance to extinction. The relation between reinforcement magnitude and response maintenance was further evaluated in the second experiment by exposing the communicative response to 20-s or 300-s access to toys or escape. Results for 2 participants suggested that this factor may alter the duration of postreinforcement pauses. PMID- 11936545 TI - Predicting the relative efficacy of three presentation methods for assessing preferences of persons with developmental disabilities. AB - Choices were presented to 9 individuals with developmental disabilities using a two-choice format. Each pair of items, selected based on prior preference assessment, was presented to each participant in three conditions (actual items, pictures of the items, and spoken-name presentation) using a reversal design. The evaluation was conducted using food items, and was then repeated using nonfood items. The participants were also given a test to measure their skills on discrimination tasks ranging in difficulty from simple to conditional discriminations. The participants' abilities to make consistent choices with food and nonfood items were predicted, with 94% accuracy, by their discrimination skills. The findings suggest that presentation methods can affect the accuracy of a choice assessment, and that the systematic assessment of basic discrimination skills can be used to predict the effectiveness of different presentation methods in this population. PMID- 11936546 TI - The effects of competing reinforcement schedules on the acquisition of functional communication. AB - The initial efficacy of functional communication training (FCT) was evaluated when problem behavior continued to produce intermittent reinforcement. Results for 2 of 3 participants showed that FCT was most effective when problem behavior was also exposed to extinction, response blocking, or both. PMID- 11936547 TI - Analysis of response covariationamong multiple topographies of food refusal. AB - This study examined the effects of sequentially introducing treatment across multiple topographies of food refusal. Treatment with nonremoval of the spoon produced an increase in food acceptance and a decrease in disruption, but expulsion of food increased. When expulsion was treated, packing of food increased. Finally, when packing was treated, all refusal behaviors remained low, and acceptance continued to occur at high and stable levels. PMID- 11936548 TI - Further evaluation of the role of protective equipment in the functional analysis of self-injurious behavior. AB - Using a procedure similar to the one described by Le and Smith (in press), we evaluated the effects of protective equipment during a functional analysis for 2 individuals who engaged in severe self-injurious behavior (SIB). Results of our analyses revealed that the use of protective equipment during functional analyses of SIB suppressed levels of responding such that a behavioral function could not be identified. PMID- 11936549 TI - Teacher acquisition of functional analysis methodology. AB - The current study examined methods for training teachers to use functional analysis methods. Teachers first received written and verbal instructions detailing attention and demand conditions. They then received training that included modeling, rehearsal, and performance feedback. Finally, probes were taken during ongoing class instruction. Results indicate that teachers acquired the skills and used them in classroom settings. PMID- 11936551 TI - A prework assessment of task preferences among adults with autism beginning a supported job. AB - A prework paired-task assessment was evaluated for identifying work preferences among 3 adults with autism beginning a supported job. When the workers began the job, choices were provided between more and less preferred tasks (determined by previous assessment). Results supported the assessment for identifying single task preferences, but did not reveal preferences of 2 workers for alternate tasks. Results are discussed in terms of evaluating other prework assessments that may reveal task-alternation preferences. PMID- 11936550 TI - Increasing social initiations in children with autism: effects of a tactile prompt. AB - An ABAB design was used to assess the effects of a tactile prompting device (i.e., a vibrating pager) as a prompt for the social initiations of 3 children with autism during free-play activities with typically developing peers. Results indicated that the tactile prompt was effective in increasing verbal initiations for all 3 children, and responses to peers' initiations were higher for 2 participants when the tactile prompt was used. Efforts to reduce the frequency of prompts while still maintaining rates of initiations were partially successful for 1 participant. PMID- 11936552 TI - Evaluation of a treatment package to reduce rapid eating. AB - We replicated and extended treatment procedures described by Lennox, Miltenberger, and Donnelly (1987) designed to reduce rapid eating. The participant was a 17-year-old girl with developmental disabilities who engaged in dangerously high rates of food ingestion. The procedure involved an adjusting differential-reinforcement-of-low-rate-responding (DRL) schedule, response blocking, and prompts. We evaluated a continuation of the treatment despite initial negative side effects that were similar to those reported by Lennox et al. Results showed that the treatment package was effective and negative side effects eventually decreased. PMID- 11936553 TI - Functional analysis of stereotypical ear covering in a child with autism. AB - We studied stereotypical ear covering in a child with autism. Results of a descriptive analysis were inconclusive but revealed a correlation between ear covering and another child's screaming. An analogue functional analysis showed that ear covering was emitted only when the screaming was present. PMID- 11936554 TI - Functional assessment and noncontingent reinforcement in the treatment of disruptive vocalization in elderly dementia patients. AB - Noncontingent reinforcement (NCR) was used as an intervention with 2 elderly dementia patients who engaged in disruptive vocalization. Several assessment procedures, including functional analysis, were conducted to identify reinforcing stimuli for use in the NCR intervention. Functional analyses and the NCR intervention were implemented in each participant's natural environment. NCR was effective in reducing disruptive vocalizations. PMID- 11936555 TI - Calcium and retinal function. AB - We survey the primary roles of calcium in retinal function, including photoreceptor transduction, transmitter release by different classes of retinal neuron, calcium-mediated regulation of gap-junctional conductance, activation of certain voltage-gated channels for K+ and Cl-, and modulation of postsynaptic potentials in retinal ganglion cells. We discuss three mechanisms for changing [Ca2+]i, which include flux through voltage-gated calcium channels, through ligand-gated channels, and by release from stores. The neuromodulatory pathways affecting each of these routes of entry are considered. The many neuromodulatory mechanisms in which calcium is a player are described and their effects upon retinal function discussed. PMID- 11936556 TI - Role of the nitric oxide/cyclic GMP/Ca2+ signaling pathway in the pyrogenic effect of interleukin-1beta. AB - Interleukin-1beta (IL-1beta) has a wide spectrum of inflammatory, metabolic, haemopoietic, and immunological properties. Because it produces fever when injected into animals and humans, it is considered an endogenous pyrogen. There is evidence to suggest that Ca2+ plays a critical role in the central mechanisms of thermoregulation, and in the intracellular signaling pathways controlling fever induced by IL-1beta and other pyrogens. Data from different labs indicate that Ca2+ and Na+ determine the temperature set point in the posterior hypothalamus (PH) of various mammals and that changes in Ca2+ and PGE2 concentrations in the cerebrospinal fluid (CSF) of these animals are associated with IL-1beta-induced fever. Antipyretic drugs such as acetylsalicylic acid, dexamethasone, and lipocortin 5-(204-212) peptide counteract IL-1beta-induced fever and abolish changes in Ca2+ and PGE2 concentrations in CSF. In vitro studies have established that activation of the nitric oxide (NO)/cyclic GMP (cGMP) pathway is part of the signaling cascade transducing Ca2+ mobilization in response to IL-1beta and that the ryanodine (RY)- and inositol-(1,4,5) trisphosphate (IP3)-sensitive pools are the main source of the mobilized Ca2+. It is concluded that the NO/cGMP/Ca2+ pathway is part of the signaling cascade subserving some of the multiple functions of IL-1beta. PMID- 11936560 TI - The recruitment of normal healthy volunteers: a review of the literature on the use of financial incentives. AB - Unresolved issues of ethical, methodological, and legal concerns in the use of normal healthy volunteers persist. Financial incentives in their recruitment offer a unique ethical dilemma because of questions surrounding payment. A review of literature was conducted to obtain research systematically examining volunteer motivation and the role offinancial incentives. The primary selection criterion was motivation and payment to volunteers; seven studies met the criterion for review. Studies that have systematically investigated volunteer motivation have found financial rewards to be an important motivator among normal healthy volunteers in their decision to participate in clinical trials. Also evident is that differences based on demographic characteristics exist in the motivation and rates of volunteerism. Ethical issues surrounding the use of normal healthy volunteers are discussed, with attention to the issue offinancial incentives (e.g., economically vulnerable volunteers, undue inducements). Regulations, guidelines, and recommendations are discussed with regard to volunteers and financial incentives. PMID- 11936558 TI - Neurobiology of nitrous oxide-induced antinociceptive effects. AB - Nitrous oxide (N2O), or laughing gas, has been used for clinical anesthesia for more than a century and is still commonly used. While the anesthetic/hypnotic mechanisms of N2O remain largely unknown, the underlying mechanisms of its analgesic/antinociceptive effects have been elucidated during the last several decades. Evidence to date indicate that N2O induces opioid peptide release in the periaqueductal gray area of the midbrain leading to the activation of the descending inhibitory pathways, which results in modulation of the pain/nociceptive processing in the spinal cord. The types of opioid peptide induced by N2O and the subtypes of opioid receptors that mediate the antinociceptive effects of N2O appear to depend on various factors including the species and/or strain, the regions of the brain, and the paradigms of behavior testing used for the experiments. Among three types of descending inhibitory pathways, the descending noradrenergic inhibitory pathway seems to play the most prominent role. The specific elements involved are now being resolved. PMID- 11936561 TI - Limited sampling strategy to predict AUC of the CYP3A phenotyping probe midazolam in adults: application to various assay techniques. AB - Midazolam clearance is used to phenotype hepatic CYP3A activity but requires multiple plasma samples following a single intravenous dose. The authors evaluated the use of a limited sampling scheme, using different assay techniques to determine the reproducibility of such a strategy in estimating midazolam AUC. Seventy-three healthy adults received midazolam as a single intravenous bolus dose. At least eight plasma samples were collected from each subject and were assayed using either LC/MS/MS or electron capture gas chromatography. Eleven subjects were randomly selected for the training set using stepwise linear regression to determine relationships between midazolam plasma concentrations and AUC. Validation of the predictive equations was done using the remaining 62 subjects. Mean percent error (MPE), mean absolute error (MAE), and root mean square error (RMSE) were calculated to determine bias and precision. Based on the training set, five models were generated with coefficients of determination ranging from 0.87 to 0.95. Validation showed that MPE, MAE, and RMSE values were acceptable for three of the models. Intrasubject reproducibility was good. In addition, training set datafrom one institution were able to predict data from the other two institutions using other assay techniques. Minimized plasma sampling mayprovide a simpler method for estimating midazolam AUC for CYP3A phenotyping. A limited sampling strategy is more convenient and cost-effective than standard sampling strategies and is applicable to more than one assay technique. PMID- 11936557 TI - Candidate RNA-binding proteins regulating extrasomatic mRNA targeting and translation in mammalian neurons. AB - In mammalian neurons, long-lasting changes in the efficacy of individual synapses depend on the synthesis of new proteins. To maintain specificity, neuronal cells have to ensure that these newly synthesized proteins accumulate at the appropriate subpopulation of synapses. One way that neurons have solved this challenge appears to be the local translation of extrasomatic mRNAs in dendrites and at postsynaptic sites. Mechanisms, which regulate the targeting, translation, and stability of dendritic mRNAs, involve an organized interaction between cis acting elements of localized transcripts and trans-acting RNA-binding proteins. The molecular identity and cellular functions of trans-acting factors that are likely to play an important role in post-transcriptional processing of extrasomatic transcripts in mammalian neurons are now being elucidated. PMID- 11936562 TI - Bioavailability and metabolism of mometasone furoate: pharmacology versus methodology. AB - The degree of systemic exposure ofter inhalation of corticosteroids is of great clinical concern. For optimum outcome, the pulmonary deposition should be sufficiently high to produce the desired anti-inflammatory effect in the lungs, whereas the plasma concentrations due to the absorption of the corticosteroid from the lung and the gut should be minimal. Recently, it has been reported that inhaled mometasone furoate has a systemic bioavailability of less than 1%, which is much lower than other corticosteroids currently available. However, critical evaluation of the study methodology and results does not support this finding. A major shortfall of the study was an insufficient analytical sensitivity, resulting in a calculated average plasma concentration profile that was entirely below the limit of quantification. These numbers were generated by replacing all concentrations below the limit of quantification byzero and then calculating an average value. This procedure can lead to erroneous results and misinterpretation. Furthermore, the potential contribution of active metabolites needs to be adequately addressed in comparisons of inhaled corticosteroids. Reliable estimates of systemic drug exposure are critical in evaluating the real safety profiles and therapeutic index for inhaled corticosteroids that are effective in treating chronic asthma. PMID- 11936564 TI - Safety and pharmacokinetics of oral voriconazole in patients at risk of fungal infection: a dose escalation study. AB - The objective of this study was to investigate the safety, tolerability, and pharmacokinetics of oral voriconazole in subjects at high risk of developingfungal infections. This was a multicenter, randomized, double-blind, double-dummy, parallel-group, dose escalation study with a fluconazole active control. Twenty-four subjects with hematological malignancies, solid tumors, or autologous bone marrow transplants were randomized to receive voriconazole 200 mg q 12 h (n = 9), voriconazole 300 mg q 12 h (n = 9), or fluconazole 400 mg OD (n = 6)for a period of 14 days. Blood samples were taken for the assessment of voriconazole pharmacokinetics in plasma on Days 1 and 14. Using a 200 mg q 12 h dosing regimen, geometric mean voriconazole peak plasma concentrations (Cmax) were 904 ng/ml on Day 1 and 2996 ng/ml on Day 14. Geometric mean voriconazole exposure, as measured by the area under the curve within a dosing interval (AUCtau), was 4044 and 20308 ng x h/ml on Days 1 and 14, respectively. On Day 1, geometric mean Cmax and AUC were 1.80- and 1.94-fold higher in subjects receiving voriconazole 300 mg q 12 h than in those receiving 200 mg q 12 h. Similarly, on Day 14, geometric mean Cmax and AUC were 1.56- and 1.80-fold greater in the high dose group. Although the confidence intervals are large, this trend suggests nonlinearity in pharmacokinetics with respect to dose as seen in healthy volunteers. The absorption of orally administered voriconazole was relatively rapid, with t(max) achieved in 1.7 to 3.0 hours. There was a mean 5.4- and 5.0 fold accumulation of voriconazole over the 14-day study period in the 200 mg and 300 mg q 12 h dose groups, respectively. Voriconazole was generally safe and well tolerated. Mild, reversible visual disturbances were the most commonly reported adverse event but were not associated with treatment discontinuation. No patient developed a breakthrough fungal infection. It was concluded that in this group of patients at risk of fungal infection, voriconazole pharmacokinetics was consistent with that reported in healthy volunteers. PMID- 11936563 TI - Effects of azimilide on heart rate and ECG conduction intervals during sinus rhythm in patients with a history of atrial fibrillation. AB - The purpose of this study was to assess the effect of azimilide, a Class III antiarrhythmic drug, on ECG conduction intervals recorded during sinus rhythm in patients with a history of symptomatic atrial fibrillation or atrial flutter. In Phase I clinical pharmacology studies, azimilide was associated with prolongation of the QT and QTc intervals on electrocardiograms recorded during sinus rhythm in normal subjects, but the effect of azimilide on the target population of patients with atrial fibrillation has not been reported in detail previously. Patients with a history of atrial fibrillation, atrial flutter, or both were randomly assigned to receive placebo or azimilide twice daily for 3 days and then qd thereafter. Azimilide doses of 50 mg, 100 mg, or 125 mg were tested. The RR, PR, QRS, QT, QTc(Bazett), and QTc(Fridericia) intervals were measured from electrocardiograms recorded at baseline and on Day 4 of test therapy. Increasing azimilide doses were associated with significant increases in the RR, QT, QTc(Bazett), and QTc(Fridericia) compared with changes in the placebo group based on the F-test for differences among treatment groups and the test for a dose response. In the azimilide 125 mg dose group, the mean change in RR was significantlygreater than the mean change in the placebo group (+61.4 ms in the azimilide 125 mg group vs. -14.1 ms in the placebo group). The mean change in QT was significantly greater in the azimilide 125 mg group than the mean change in the placebo group (+44.2 ms in the azimilide 125 mg group vs. -1.0 ms in the placebo group). The mean change in QTc using both correction methods was significantly greater in the azimilide 125 mggroup than the mean change in the respective placebo group: QTc(Bazett) +31.6 ms in the azimilide 125 mg group versus +2.1 ms in the placebo group and QTc(Fridericia) +35.8 in the azimilide 125 mg group versus +1.0 ms in the placebo group. It was concluded that in patients with a history of atrial fibrillation or flutter, azimilide was associated with statistically significant increases in RR, QT, QTc(Bazett), and QTc(Fridericia) when patients were in sinus rhythm. PMID- 11936565 TI - Comparative target site pharmacokinetics of immediate- and modified-release formulations of cefaclor in humans. AB - Optimal dosing of beta-lactam antibiotics aims at maximizing the time at which drug levels in the interstitial space fluid (ISF)--the fluid that surrounds the causative microorganisms at the target site--exceed the minimal inhibitory concentration (MIC). One potentially attractive strategy to achieve this goal is to administer antibiotics as oral sustained-release formulations. The present study was designed to test the hypothesis that sustained-release formulations could lead to a more suitable pharmacokinetic profile in the ISF at the relevant target site. For this purpose, time versus cefaclor concentration profiles attained in the ISF were measured following administration of two formulations, an immediate- (500 mg IR) and a modified-release formulation in two different doses (500 mg MR and 750 mgMR) in a three-way crossover study of healthy male volunteers (n = 12). For the measurement of unbound cefaclor concentrations in the ISF of human skeletal muscle, the in vivo microdialysis technique was employed. For all three formulations, unbound cefaclor concentration in the ISF closely followed individual plasma concentration profiles in a dose-dependent pattern, with ISF to unbound plasma ratios ranging from 0.67 to 0.73. The mean residence time was found to be significantly longer for the MR formulations versus the IR formulation. The data of the present study indicate that time above MIC values at the target site can be substantially prolonged if an antibiotic is administered as a sustained-release product. PMID- 11936566 TI - Pharmacokinetic study of infusional valspodar. AB - The pharmacokinetics of valspodar (PSC 833), a selective second-generation P glycoprotein modulator, was evaluated as part of a Phase I study to modulate paclitaxel therapy in 15 patients with refractory malignancies. Valspodar was given intravenously at 1.42 mg/kg/h for 2 hours, followed by a 27-hour continuous infusion at 0.42 mg/kg/h. Serial blood samples were obtained after intravenous infusion of valspodar and paclitaxel. Valspodar disposition was best described by a linear two-compartment model. The median (range) valspodar clearance was 0.40 ml/min/kg (0.07-1.40 ml/min/kg). The 20-fold interpatient variability in valspodar clearance was not correlated with age, body weight, orgender but might be associated with coadministered medications that were metabolized via cytochrome P450 3A-mediated elimination. Valspodar whole-blood concentrations were maintained above the target threshold of 1000 ng/ml for a median of 32 hours. The pharmacokinetic model generated from this study allows for application in future studies to optimize the use of valspodar. PMID- 11936559 TI - Glutamate receptor genes: susceptibility factors in schizophrenia and depressive disorders? AB - Schizophrenia, depression, and bipolar disorder are three major neuropsychiatric disorders that are among the leading causes of disability and have enormous economic impacts on our society. Although several neurotransmitter systems have been suggested to play a role in their etiology, we still have not identified any gene or molecular mechanism that might lead to genetic susceptibility for or protection against these neuropsychiatric disorders. The glutamatergic receptor system, and in particular the N-methyl-D-aspartate (NMDA) receptor complex, has long been implicated in their etiology. I review the current molecular evidence that supports a critical role for the glutamatergic receptor system in schizophrenia and the potential involvement of this receptor system in depression and bipolar disorder. It is likely that mutations in glutamate receptor genes might alter the risk of developing one of these disorders. Potential future research directions designed to identify these mutations and to elucidate their effect on mental health will be discussed. PMID- 11936567 TI - Effect of food on the oral bioavailability of didanosine from encapsulated enteric-coated beads. AB - The objective of this study was to assess the effect of food and timing of meals on the bioavailability of didanosine from encapsulated enteric-coated beads. Four different independent, open-label, single-dose, randomized, crossover studies were conducted in healthy subjects (n = 20-30). Didanosine (400 mg) was given concomitantly with a high-fat meal, light meal, yogurt, and applesauce. In addition, didanosine was given 1, 1.5, 2, and 3 hours before and 2 hours after a light meal. Statistical comparison with fasting conditions was made using the equivalence approach for Cmax (70%-143%) and AUC (80%-125%). The high-fat meal, light meal, yogurt, and applesauce decreased the Cmax by 46%, 22%, 30%, and 24%, respectively, and lowered the AUC by 19%, 27%, 20%, and 18%, respectively; statistical analyses indicated an indeterminate food effect, except for the Cmax for the high-fat meal. For 1 hour before meal, Cmax and AUC were lower by 15% and 24% and, for 2 hours after meal, were lower by 15% and 10%, respectively. There was an indeterminate food effect for 1 hour before the meal treatment; in addition, 2 hours after the meal, treatment approached statistical equivalence, missing narrowly on the lower bounds. For 1.5, 2, and 3 hours before meal treatments, Cmax values were unchanged, but AUC was lower by 10%, 4%, and 0%, respectively; lack of food effect was observed for all three treatments. Across studies, median time to Cmix ranged from 1.67 to 2.67 hours but was delayed by 2.5 to 3 hours with high-fat and light meals compared to fasting conditions. The half-life of didanosine was 1.5 to 2 hours. It was concluded that the bioavailability of didanosine from encapsulated enteric-coated beads was reduced by approximately 20% to 25% with food, which can be circumvented by taking didanosine on an empty stomach. The clinical significance of such moderate reductions in didanosine exposure with food, especially as part of a highly active antiretroviral therapy, is not clear. PMID- 11936568 TI - Pharmacokinetics and safety of galantamine in subjects with hepatic impairment and healthy volunteers. AB - The objective of this study was to compare the pharmacokinetics and safety of galantamine in subjects with hepatic impairment with those in healthy subjects. This was an open-label study in which a single oral 4-mg dose of galantamine was administered to volunteers with mild (Child-Pugh score of 5-6, n = 8), moderate (Child-Pugh score of 7-9, n = 8), or severe hepatic impairment (Child-Pugh score of 10-15, n = 1) and to healthy, matched control subjects (n = 8). Galantamine pharmacokinetics and safety (adverse events, laboratory test results, electrocardiograms, vital signs, and cardiac events) were assessed over 6 days after administration of galantamine. The pharmacokinetic parameters of galantamine were similar in subjects with mild hepatic impairment compared with healthy controls. Compared with the healthy control group, subjects with moderate hepatic impairment showed relative increases in the area under the plasma concentration curve from zero to infinity (AUC0-infinity) (+33%) and terminal half-life (t1/2) (+30%) (p = 0.051 and p = 0.003, respectively), a 23% relative decrease in total plasma clearance (p = 0.061), and a small but significant relative increase in the fraction of free plasma galantamine (p = 0.009). Galantamine was well tolerated by all subjects. There were no serious adverse events (AEs) or premature withdrawals from the study because of AEs. Reported AEs were headache (three cases), nausea (one case), and paresthesia (one case). There were no clinically relevant changes in clinical laboratory findings, vital signs, and electrocardiograms. Low patient recruitment (n = 1) precluded statistical analysis of galantamine pharmacokinetics and safety in severe hepatic impairment. It was concluded that the pharmacokinetics of galantamine in subjects with mild hepatic impairment was similarto those in healthy subjects. In subjects with moderate hepatic impairment, galantamine clearance was decreased by approximately 23% compared with normal volunteers. Galantamine was also well tolerated and appeared to be safe in subjects with mild ormoderate hepatic impairment. Based on the study results, it appears that it would not be necessary to adjust doses of galantamine during administration to subjects with mild hepatic impairment. In subjects with moderately impaired hepatic function, dose titration should proceed cautiously. Unfortunately, difficulties with patient recruitment did not allow adequate assessment of the safety of galantamine in subjects with severe hepatic impairment in this study. Therefore, the use of galantamine in subjects with severe hepatic impairment is not recommended. PMID- 11936569 TI - Is it really OK to take this with food? Old interactions with a new twist. AB - In response to consumers' increased interest in preventive health care, the food industry is producing a variety of foods fortified with calcium, iron, and other minerals and vitamins. This well-meaning idea of food fortification is troubling in the context of clinical pharmacology. The recommended Food and Drug Administration (FDA) meal used in food-drug interaction studies is a high-fat, high-calorie meal with little nutritive value. While some drugs may appear to be safe when taken with food, this may not be true when fortified foods are considered. The mechanisms causing drug-fortified food interactions are the some well-known mechanisms that cause other drug-mineral interactions. Certain drugs may exhibit decreased absorption due to chelation and adsorption. Other drugs may have decreased absorption or increased excretion due to changes in gastric and/or urinary pH. The results of such interactions may be clinically insignificant or severe, including treatment failure, frequent dose changes, antibiotic resistance, and increased morbidity and mortality. Revisions of current regulatoryguidelines are necessary to take into account this potentially major source of "new" drug interactions. PMID- 11936570 TI - A study of the interaction potential of azithromycin and clarithromycin with atorvastatin in healthy volunteers. AB - Atorvastatin is a common option among the HMG-CoA reductase inhibitors for the treatment of lipid disorders because of its excellent lipid-lowering efficacy and overall safety profile. Although these agents can rarely cause rhabdomyolysis by themselves, macrolides, among other agents, have been demonstrated to increase the likelihood of this via inhibition of CYP metabolism of the lipid agent. This study investigated the potential for azithromycin and clarithromycin to inhibit the metabolism of atorvastatin. Although there was no interaction between azithromycin and atorvastatin, clarithromycin did have a significant effect on atorvastatin pharmacokinetic parameters. When coadministered, clarithromycin raised subject exposure (AUC24) by 82% and peak plasma concentrations by 56%. These data suggest that while azithromycin appears to be safe to coadminister with atorvastatin, clarithromycin should be avoided in patients taking this and similarly metabolized HMG-CoA inhibitors. PMID- 11936571 TI - Steady-state pharmacokinetics and tolerability of modafinil administered alone or in combination with dextroamphetamine in healthy volunteers. AB - The potential for a drug-drug interaction between modafinil and dextroamphetamine, each at steady state, was investigated in an open-label, randomized, single-period studyin 32 healthy male and female volunteers. All subjects received modafinil orally once daily for 28 days (200 mg on Days 1-7; 400 mg on Days 8-28). On Days 22 to 28, half of the subjects also received dextroamphetamine (20 mg) orally 7 hours after modafinil. Samples for pharmacokinetic (PK) profiling were obtained on Days 21 and 28. The mean changes in PK parameters for modafinil and its two circulating metabolites between the two groups were not statistically significantly different, except Cmax for modafinil acid. Adverse events obtained in the two groups were similar and mild or moderate in nature. The results indicate that administration of low-dose dextroamphetamine in this dosing regimen does not alter the steady-state pharmacokinetics of modafinil. The combination has a similar tolerability profile as modafinil alone. PMID- 11936572 TI - Lack of bioequivalence of ciprofloxacin when administered with calcium-fortified orange juice: a new twist on an old interaction. AB - Fluoroquinolones are known to interact with drugs containing multivalent ions. Current Food and Drug Administration (FDA) labeling states that ciprofloxacin and most other fluoroquinolones are safe to be given with food and dietary calcium but not calcium supplements. Although many of the currently marketed calcium fortified foods have calcium contents that usually exceed those in dietary calcium sources, it is unclear whether they represent a risk for less than optimal absorption of fluoroquinolones, which may result in subsequent clinical failures due to lack of bacterial eradication and antibiotic resistance. The purpose of this three-way, randomized, crossover study was to characterize and compare the bioequivalence of single doses of oral ciprofloxacin in 15 healthy volunteers when administered with water, concurrently with orange juice, and concurrently with calcium-fortified orange juice. Compared to the control arm, the Cmax of ciprofloxacin significantly decreased when it was given with orange juice (23%, p = 0.001) and with calcium-fortified orange juice (41%, p < 0.001). Twenty-four-hour ciprofloxacin AUCs were also decreased for both forms of the orange juice (22% [p < 0.001] and 38% [p < 0.001], respectively). When compared to each other, neither of the orange juice regimens were bioequivalent to each other, with the Cmax and AUC for the fortified form being 22% (p = 0.005) and 21% (p = 0.015) lower than those of the nonfortified form. By FDA standards, although ciprofloxacin is marginally bioequivalent when administered with orange juice, it is not when it is administered with calcium-fortified orange juice. The changes in Cmax and AUC have the potential to significantly decrease clinical efficacy and promote antibiotic resistance. Not warning patients about potential food-drug interactions with fortified foods may be a major unrealized and unstudied inadvertent source of clinical failures and resistance trends with fluoroquinolones. PMID- 11936573 TI - Human biokinetics of injected bismuth-207. AB - A healthy male volunteer received an intravenous injection of 207Bi as citrate. Levels of the tracer in blood and in excretion samples, and its retention and distribution within the body, were investigated by appropriate radioactivity measurements. Levels in blood fell very rapidly, with only 1% of the injection remaining at 7 h and only ca. 0.1% at 18 days. There was rapid initial excretion, with 55% lost during the first 47 h, principally in urine; however, longer-term losses were much slower and 0.6% remained in the body at 924 days, when the contemporary rate of loss implied a half-life of 1.9 years. Integration of the retention pattern suggested that steady exposure to bismuth compounds could lead ultimately to a body content of approximately 24 times the daily systemic uptake. The largest organ deposit was in the liver, which after 3 days contained ca. 60% of the contemporary whole body content, consistent with reports of hepatotoxicity. These findings differ markedly from the metabolic model for bismuth proposed by the International Commission on Radiological Protection, which envisages a terminal half-life in the body of only 5 days and kidney as the site of highest deposition. PMID- 11936574 TI - Parasuicidal poisoning treated in a Greek medical ward: epidemiology and clinical experience. AB - OBJECTIVE: To study the epidemiology of acute poisoning patients presenting to an acute medical service ward in a Greek hospital between January 1998 and December 2000. DESIGN: Prospective case series. RESULTS: A total of 273 patients with self poisoning were included in the study. This represented 3.8% of the overall admissions to the unit. The mean age of patients was 33, the most frequent age group being that aged 20-30 years (36.2% of total) with a male-to-female ratio of 1:1.97. Sixty per cent of patients was admitted within 4 h. Those from urban areas comprised 76.2% and 23.8% from rural areas. The most frequently ingested agents were psychopharmaceuticals (37.4%) and analgesics/anti-rheumatics (32.6%). Pesticides (7.7% of total) were most frequently used by patients coming from rural areas (32.3% of patients from rural areas). Alcohol was included in the overdose in 8.4%. Of the patients, 16.2% had a previous history of overdose. In this case series, psychiatric assessment suggested that 52% of the patients had a formal psychotic diagnosis, 21% had personality disorder and 27% had taken an overdose in response to stress. The most frequently documented precipitating factors were family problems and disputes (37%). Unusually, the seasonal distribution in these patients suggested a peak in summer (37.5% of presentations) with lower numbers in spring (30.2%), autumn (17.7%) and winter (14.6%). Of the patients, 23.7% presented in July. A total of 73.5% of patients was conscious, 16.4% was somnolent, 4.5% was in precoma and 5.6% was in coma (GCS <8). Patients who received antidotal therapy comprised 17.9%. Evidence of hepatic dysfunction was observed in 8.9% of patients and renal dysfunction in 3.6%. Extracorporeal techniques for drug removal (hemodialysis and hemoperfusion) were used in 2.2% of patients. Intensive care therapy was required in 11.4% of patients. The mean overall hospitalization time was 3.3 days. The mortality rate was 2.9%. CONCLUSIONS: This study shows that the epidemiology of self-harm by overdose in Greece is significantly different in terms of the seasonal presentation from other parts of Europe. The agents ingested and other features are similar to northern Europe. Psychiatric diagnoses are more common in our group than in those reported from northern Europe. PMID- 11936575 TI - Vitamin C ameliorates fluoride-induced embryotoxicity in pregnant rats. AB - Oral administration of sodium fluoride (40 mg/kg body weight) from day 6 to 19 of gestation caused, as compared to control, significant reductions in body weight, feed consumption, absolute uterine weight and number of implantations. Significantly higher incidence of skeletal (wavy ribs, 14th rib, <6 sternal centre, dumbell-shaped second and fifth sternebrae, incomplete ossification of skull and thickening of tibia) and visceral (subcutaneous haemorrhage) abnormalities were also observed in NaF-treated dams than that of control. Oral administration of vitamin C (50 mg/kg body weight) and vitamin E (2 mg/0.2 ml olive oil/animal/day) from day 6 to 19 of gestation along with NaF significantly ameliorates NaF-induced reductions in body weight, feed consumption, absolute uterine weight (only with vitamin E treatment) and number of implantations. As compared with NaF-treated alone, the total percentage of skeletal and visceral abnormalities were significantly lowered in fluoride plus vitamin C-treated animals. Vitamin E was less effective. These findings suggest that vitamin C significantly reduced the severity and incidence of fluoride-induced embryotoxicity in rats. PMID- 11936576 TI - Carbendazim-induced haematological, biochemical and histopathological changes to the liver and kidney of male rats. AB - Carbendazim is a systemic broad-spectrum fungicide controlling a wide range of pathogens. It is also used as a preservative in paint, textile, papermaking and leather industry, as well as a preservative of fruits. In the present study, carbendazim was administered at 0, 150, 300 and 600 mg/kg per day doses orally to male rats (Rattus rattus) for 15 weeks. At the end of the experiment, blood samples, liver and kidney tissues of each animal were taken. Serum enzyme activities, and haematological and biochemical parameters were analysed. In toxicological tests, 600 mg/kg per day doses of carbendazim caused an increase of albumin, glucose, creatinine and cholesterol levels. Also, at the same doses, white blood cell and lymphocyte counts decreased. However, mean cell hemoglobin and mean cell hemoglobin concentrations increased. Histopathological examinations revealed congestion, an enlargement of the sinusoids, an increase in the number of Kupffer cells, mononuclear cell infiltration and hydropic degeneration in the liver. At the highest doses, congestion, mononuclear cell infiltration, tubular degeneration and fibrosis were observed in the kidney tissue. These results indicate that 300 and 600 mg/kg per day carbendazim affected the liver and kidney tissue and caused some changes on haematological and biochemical parameters of rats. PMID- 11936577 TI - Platelet dopamine receptors and oxidative stress parameters as markers of manganese toxicity. AB - The present study has been undertaken to investigate whether neurotoxic effects of manganese (Mn) are reflected in platelets in rats to monitor the usefulness of platelet as peripheral model. Exposure of rats to Mn (10 or 15 mg/kg bw, i.p.) for 45 days caused a significant increase in membrane fluidity as evidenced by decrease in fluorescence polarisation in platelets (11% and 14%) and striatum (9% and 13%). These rats exhibited a significant increase in superoxide dismutase activity both in platelets (24% and 37%) and striatum (31% and 42%), respectively, in comparison to controls. Exposure of rats to Mn for 45 days (15 mg/kg bw, i.p.) caused a significant decrease in reduced glutathione content (platelets 20%, striatum 24%) and catalase activity (platelets 35%, striatum 44%) compared to control rats. Rats exposed to Mn (10 or 15 mg/ kg bw, i.p.) for 15 days exhibited a significant increase in dopamine receptors both in platelets (55% and 40%) and striatum (38% and 31%). The results suggest that exposure to Mn may alter the membrane functions and impair the anti-oxidant defense mechanism both in platelets and brain. The study also suggests that dopaminergic mechanisms are impaired following Mn exposure and such changes are reflected in platelets. Interestingly, parallel changes both in striatum and platelets, as observed in the present study, strengthen the usefulness of platelets as a peripheral neuronal model. PMID- 11936578 TI - Captopril inhibits the pulmonary toxicity of paraquat in rats. AB - Paraquat (PQ) is a herbicide that is very toxic to all living organisms. It generates free radicals and leads to acute or chronic lung injury. Free radicals are often associated with fibrogenesis, which occurs in various disease states. The purpose of this study was to determine whether captopril prevents paraquat toxicity in lung tissue. Paraquat alone increased the level of lipid peroxidation (LPO) and the activity of superoxide dismutase (SOD) after 4, 12, 24 and 72 h of administration. Also, the level of hydroxyproline showed an increase after 24 h of paraquat administration. However, paraquat also decreased the level of glutathione (GSH) and the activity of glutathione peroxidase (GSH-Px). Captopril (50 mg/kg i.p.) and paraquat were simultaneously injected (40 mg/kg i.p.), and the captopril injection 1 h after paraquat ameliorated the biochemical toxicity induced by paraquat. This was evidenced by a significant reduction in LPO and balancing the endogenous antioxidant capacity by normalizing the activities of SOD and GSH-Px and the GSH content in the lung tissue. Moreover, captopril injection prevented the increase of hydroxyproline content as an index of lung fibrosis. From these results, the beneficial effects of captopril on paraquat toxicity appear to be through enhancement of the endogenous antioxidant system preventing the lung fibrosis. PMID- 11936579 TI - Protective effects of Emblica officinalis Gaertn. in cyclophosphamide-treated mice. AB - Cyclophosphamide (CP) is one of the most popular alkylating anticancer drugs in spite of its toxic side effects including immunotoxicity, hematotoxicity, mutagenicity and a host of others. The present study was undertaken to assess the protective effects of total aqueous extract of a medicinal plant, Indian gooseberry (Emblica officinalis Gaertn.) in mice treated with CP. These protective effects were studied on immunological parameters and kidney and liver antioxidants. Plant extract treatment at a dose of 100 mg/kg body weight per os (p.o.) for 10 days resulted in the modulation of these parameters in normal as well as CP (50 mg/kg)-treated animals. Plant extract in particular was very effective in reducing CP-induced suppression of humoral immunity. Plant extract treatment in normal animals modulated certain antioxidants of kidney and liver. In CP-exposed animals, plant pretreatment provided protection to antioxidants of kidney. Not only were the reduced glutathione levels significantly (p<0.001) increased but plant extract treatment resulted in restoration of antioxidant enzymes in CP-treated animals. It is suggested that E. officinalis or its medicinal preparations may prove to be useful as a component of combination therapy in cancer patients under CP treatment regimen. PMID- 11936580 TI - Diepoxybutane and mitomycin C toxicity is associated with the induction of oxidative DNA damage in sea urchin embryos. AB - Diepoxybutane (DEB)- and mitomycin C (MMC)-associated toxicity was investigated in embryos from the sea urchin (SU) species Sphaerechinus granularis. DEB- and MMC-induced toxicity resulted in S. granularis embryos and larvae at concentrations ranging 10(-5) to 10(-4) M DEB, and 3 x 10(-6) to 3 x 10(-5) M MMC, in terms of larval malformations, developmental arrest and mortality. The formation of DNA oxidative damage, 8-hydroxy-2'-deoxyguanosine (8-OHdG) was measured in DEB- and in MMC-exposed embryos (at gastrula stage). A dose-dependent increase in 8-OHdG levels was observed that was significantly correlated with DEB and MMC-induced developmental defects. The results lend further support to the body of evidence associating both DEB and MMC toxicity with oxidative stress, including DNA oxidative damage. PMID- 11936581 TI - Repeated measurements of aldicarb in blood and urine in a case of nonfatal poisoning. AB - A nonfatal case of poisoning involving aldicarb, an extremely toxic carbamate pesticide, is presented. A 39-year-old female ingested an unknown amount of aldicarb, together with alprazolam and sertraline. On admission to ICU (T0), she displayed marked cholinergic symptoms and a deep coma. The patient was given pralidoxime and atropine. Her condition gradually improved on days 2 and 3 and she was discharged at T0+80 h. Aldicarb was assayed by high-performance liquid chromatography on 21 blood and 8 urine samples successively taken during hospitalization. At the same time, serum pseudocholinesterase activity was followed on 21 successive samples. Blood aldicarb level was 3.11 microg/mL at T0 and peaked at T0+3.5 h (3.22 microg/mL), then followed a two-slope decay with a terminal half-life of ca. 20 h. Aldicarb was detected in all urine samples (peak level: 6.95 microg/mL at T0+31.5 h) and was still present at the time of discharge. Serum pseudo-cholinesterase activity remained low (< or = 10% of normal) until the 30th hour then rapidly increased and returned to normal after the 60th hour. The patient's clinical picture closely followed blood aldicarb levels and serum pseudo-cholinesterase activities. To our knowledge, this is the first report of an aldicarb poisoning documented by repeated measurements of the drug in the intoxicated person. PMID- 11936582 TI - New biotechnologies: spin-off on health and society. AB - New biotechnologies and the new biology deriving from them are having a revolutionary impact on economy and society and are consequently transforming the role of researchers, which is changing continually to meet the competence required. The advances in human genetics on the other hand make it necessary to look for different approaches and new rules in bioethics. Comprehensive views and careful considerations are therefore needed in order that this new biology may have a positive impact on health, being respectful of the social and ethical principles of human beings. PMID- 11936583 TI - The prognostic value of hormone receptor detection by enzyme immunoassay and immunohistochemistry: a prospective study in patients with early breast cancer. AB - The main reason to determine estrogen (ER) and progesterone receptors (PR) in breast cancer is their predictive value for the response to endocrine therapy. In addition, ER and PR are often used as prognostic indicators. Enzyme immunoassay (EIA) and immunocytochemical assay (ICA) are two methods for determining ER and PR. These two methods have not been compared with each other in relation to clinical endpoints. In the present study we prospectively evaluated the prognostic value of ER and PR as determined by ICA and EIA in 223 and 207 patients, respectively, with early breast cancer. ER was positive in approximately 77% of patients, while PR was positive in approximately 65% of patients. The proportion of potential agreement beyond chance between EIA and ICA was 0.58 and 0.65 for ER and PR, respectively. The median follow-up was 86 months. Both ER and PR appeared to be weak prognostic factors. There were no differences in prognostic value according to the time point of analysis or cutoff value chosen, nor were there any differences in the prognostic value of hormone receptors detected by ICA or EIA. Both methods appear to be equivalent in terms of qualification and prognostic value. PMID- 11936584 TI - Prognostic significance of plasminogen activator inhibitor-1 in breast cancer, with special emphasis on locoregional recurrence-free survival. AB - The independent prognostic value of protease uPA and its inhibitor PAI-1 for survival in breast cancer patients is firmly established. However, there is very little data on the prognostic value of serine proteases and their inhibitors for locoregional recurrence in breast cancer. The prognostic value of PAI-1 for local control in a group of 766 patients treated at our institute with either breast conserving treatment or modified radical mastectomy was evaluated. The locoregional recurrence-free survival (LRFS) of patients with PAI-1 values above the median value was significantly worse than that of patients with PAI-1 values below the median value (log-rank; p=0.0078). In multivariate analysis PAI-1 levels proved to be of independent statistical significance for LRFS (p=0.0401, relative risk 2.28, 95% confidence interval 1.04-5.02). The independent prognostic value of PAI-1 for metastasis-free survival and overall survival was also confirmed. In addition, our data suggest that PAI-1 antigen levels in tumor tissue might be of prognostic value for survival after locoregional recurrence (log-rank; p=0.0618). According to our findings, PAI-1 levels could be used as a biological marker that could facilitate the identifation of patients with a higher risk of local relapse already at the time of primary treatment. These patients should then be offered more aggressive treatment. PMID- 11936585 TI - TP53 mutations in human meningiomas. AB - Overexpression of p53 has been reported to play a role in the development of neoplasms of the central nervous system. Meningiomas are generally benign intracranial tumors originating from the meninges. Overexpression of the p53 protein in meningiomas and an association with histological type and recurrence has been reported. Mutation of the TP53 gene leads to a more stable p53 protein in quantities high enough for detection by immunohistochemistry. In the search for these mutations the core domain of the TP53 gene of meningiomas has been analyzed. Only a very low incidence of mutations was reported. The apparent discordance between overexpression of p53 protein and TP53 gene mutations may be explained by mutations located outside the core domain. This issue was addressed in the present study. All 11 exons of 17 meningiomas were analyzed for DNA alterations by PCR single-strand conformation polymorphism (PCR-SSCP) analysis with subsequent sequencing. PCR-SSCP analysis showed a various number of band shifts and nucleotide alterations, caused either by alterations in the flanking introns or common polymorphisms (codon 36 and 72). The allele frequencies of the polymorphisms found in this small population of tumors resemble the frequencies reported in the literature. In addition, three nucleotide changes located in introns 2, 3 and 7 were found in 11, 3 and 4, respectively, of 17 specimens. Based on this study and on reports by others we conclude that it is not very likely that TP53 mutations are involved in the etiology of meningiomas. PMID- 11936586 TI - Telomerase activity in breast cancer in Western India (Gujarat). AB - The purpose of this study was to examine the association of telomerase activity with clinical and histopathological prognostic variables in primary breast cancer (n=64). Telomerase activity in breast cancer was also compared with that in benign (n=10) and non-malignant tissues (n=8; post-lumpectomy tissues histopathologically defined as containing no residual tumor). The parameter was assessed using the Telomerase PCR ELISA kit. Values above OD 0.120 were considered positive. Estrogen and progesterone receptors (ER and PgR) were assayed by the dextran-coated charcoal method and levels >10 fmol/mg cytosol protein were taken as positive. Telomerase activity was detected in 20% and 50% of the patients with benign lesions and primary breast cancer, respectively, and in 50% of post-lumpectomy breast tissues histopathologically defined as containing no residual tumor. Telomerase activity was present in all stages of breast carcinoma and showed a significant inverse correlation with lymph node status (p=0.006), lymphatic invasion (p=0.035) and necrosis (p=0.033). Moreover, when stage II patients were grouped according to nodal involvement, a trend towards significance was observed (p=0.055). No correlation was observed with ER and PgR. The results of our study suggest that telomerase activity might be associated with the presence of cancer cells. Furthermore, telomerase activation may occur early in breast cancer and may be periodically downregulated during subsequent tumor progression. PMID- 11936587 TI - A low-density DNA microarray for analysis of markers in breast cancer. AB - Breast cancer remains a major cause of death in women from Western countries. In the near future, advances in both nucleic acids technology and tumor biology should be widely exploited to improve the diagnosis, prognosis, and outcome prediction of this disease. The DNA microarray, also called biochip, is a promising tool for performing massive, simultaneous, fast, and standardized analyses of multiple molecular markers in tumor samples. However, most currently available microarrays are expensive, which is mainly due to the amount (several thousands) of different DNA capture sequences that they carry. While these high density microarrays are best suited for basic studies, their introduction into the clinical routine remains hypothetical. We describe here the principles of a low-density microarray, carrying only a few hundreds of capture sequences specific to markers whose importance in breast cancer is generally recognized or suggested by the current medical literature. We provide a list of about 250 of these markers. We also examine some potential difficulties (homologies between marker and/or variant sequences, size of sequences, etc.) associated with the production of such a low-cost microarray. PMID- 11936588 TI - Soluble selectin levels in patients with lung cancer. AB - Increased expression of selectins has been found on endothelial cells of venules and capillaries in the tumor stroma of non-small cell lung cancer, suggesting their functional role in the process of chemotaxis for tumor cells. The present study was aimed at analyzing the role of both soluble (s)P-selectin and sE selectin levels in association with clinico-pathological variables in 116 patients with lung cancer, 38 patients with benign diseases and 59 healthy donors. The results obtained showed that sP-selectin and sE-selectin levels were higher in patients with lung cancer compared to normal donors (p<0.02 and p<0.005, respectively). No differences were observed among patients with various benign diseases for both selectins. Increased levels of sP-selectin and sE selectin were significantly associated with squamous lung cancer at late stages (p<0.05), but not adenocarcinoma. Both sP- and sE-selectin were independently related to the stage of squamous lung cancer by stepwise regression analysis (p<0.02 and p<0.03, respectively), while only sE-selectin was independently related to the presence of distant metastasis in the same histotype (p<0.02). These results suggest that measurement of plasma soluble selectins might represent a useful laboratory parameter in the management of patients with squamous lung cancer. PMID- 11936589 TI - Influence of 5-fluorouracil and folinic acid on interleukin-18 production in colorectal cancer patients. AB - AIMS AND BACKGROUND: This study was carried out to evaluate the IL-18 blood concentrations of operated colorectal cancer patients and their possible variation in response to combination chemotherapy with 5-fluorouracil (5-FU) and folinic acid. METHODS: IL-18 levels were assayed in sera of 18 healthy donors and 18 surgical colorectal cancer patients before and after adjuvant chemotherapy with 5-fluorouracil and folinic acid. An ELISA kit for human IL-18 was used for the assay. RESULTS: Colorectal cancer patients showed significantly higher baseline levels of IL-18 than healthy donors (p<0.005). Furthermore, serum IL-18 levels increased significantly with respect to baseline in patients receiving adjuvant chemotherapy (p<0.005). CONCLUSIONS: This study suggests that treatment with 5-fluorouracil and folinic acid may provoke an increase in IL-18 serum levels in colorectal cancer patients. This increase may help to explain the efficacy of adjuvant chemotherapy with 5-FU in colorectal cancer. PMID- 11936590 TI - Reference levels of the tumor markers carcinoembryonic antigen, the carbohydrate antigens 19-9 and 72-4, and cytokeratin fragment 19 using the Elecsys Relecsys 1010 analyzer in a normal population in Kuwait. The importance of the determination of local reference levels. AB - The tumor markers CEA, CA 19-9, CA 72-4 and CYFRA 21-1 were analyzed in a group of apparently healthy subjects (n=232) in Kuwait using the Elecsys Relecsys 1010 analyzer. The distribution of the tumour marker levels was analyzed separately in Kuwaitis (n=103), non-Kuwaitis (n=129), smokers (n=68), non-smokers (n=164), males (n=138) and females (n=94). The distribution of CEA was significantly different in Kuwaitis vs. non-Kuwaitis in the total population (p=0.033) and in non-smokers (p=0.049); in males vs. females in the total population (p<0.0001) and in non-smokers (p=0.0002); and in smokers vs. non-smokers in the total population (p<0.0001) using the non-parametric Mann-Whitney U test. None of the other tumour markers showed significant differences in the subgroups. The upper reference level was defined as the 95th percentile of the normal values in each group. A higher reference level of CEA was observed in smokers (vs. non-smokers) in the total population. Also higher reference levels of CEA were observed in males (vs. females) both in the total population and in non-smokers. In the total population the respective reference levels were: CEA: 4.4 microg/L, CA 19-9: 35 kU/L, CA 72.4: 2.4 kU/L, and CYFRA 21.1: 2.1 microg/L. These results were compared with data in the kit inserts and literature data. The impact of 95th percentiles in a local heterogeneous population is discussed. PMID- 11936591 TI - Genomics and proteomics meet diagnostic hematopathology. PMID- 11936592 TI - Some considerations on reading research findings displayed in a 2x2 contingency table. PMID- 11936593 TI - Information technology in biomedicine: maturational insights. PMID- 11936594 TI - Iterative normalization of cDNA microarray data. AB - This paper describes a new approach to normalizing microarray expression data. The novel feature is to unify the tasks of estimating normalization coefficients and identifying control gene set. Unification is realized by constructing a window function over the scatter plot defining the subset of constantly expressed genes and by affecting optimization using an iterative procedure. The structure of window function gates contributions to the control gene set used to estimate normalization coefficients. This window measures the consistency of the matched neighborhoods in the scatter plot and provides a means of rejecting control gene outliers. The recovery of normalizational regression and control gene selection are interleaved and are realized by applying coupled operations to the mean square error function. In this way, the two processes bootstrap one another. We evaluate the technique on real microarray data from breast cancer cell lines and complement the experiment with a data cluster visualization study. PMID- 11936596 TI - A data-hiding technique with authentication, integration, and confidentiality for electronic patient records. AB - A data-hiding technique called the "bipolar multiple-number base" was developed to provide capabilities of authentication, integration, and confidentiality for an electronic patient record (EPR) transmitted among hospitals through the Internet. The proposed technique is capable of hiding those EPR related data such as diagnostic reports, electrocardiogram, and digital signatures from doctors or a hospital into a mark image. The mark image could be the mark of a hospital used to identify the origin of an EPR. Those digital signatures from doctors and a hospital could be applied for the EPR authentication. Thus, different types of medical data can be integrated into the same mark image. The confidentiality is ultimately achieved by decrypting the EPR related data and digital signatures with an exact copy of the original mark image. The experimental results validate the integrity and the invisibility of the hidden EPR related data. This newly developed technique allows all of the hidden data to be separated and restored perfectly by authorized users. PMID- 11936595 TI - Fetal lung maturity analysis using ultrasound image features. AB - This pilot study was carried out to find the feasibility of analyzing the maturity of the fetal lung using ultrasound images. Data were collected from normal pregnant women at intervals of two weeks from the gestation age of 24 to 38 weeks. Images were acquired at two centers located at different geographical locations. The total data acquired consisted of 750 images of immature and 250 images of mature class. A region of interest of 64 x 64 pixels was used for extracting the features. Various textural features were computed from the fetal lung and liver images. The ratios of fetal lung to liver feature values were investigated as possible indexes for classifying the images into those from mature (reduced pulmonary risk) and immature (possible pulmonary risk) lung. The features used are fractal dimension, lacunarity, and features derived from the histogram of the images. The following classifiers were used to classify the fetal lung images as belonging to mature or immature lung: nearest neighbor, k nearest neighbor, modified k-nearest neighbor, multilayer perceptron, radial basis function network, and support vector machines. The classification accuracy obtained for the testing set ranges from 73% to 96%. PMID- 11936597 TI - Fractal analysis in the detection of colonic cancer images. AB - The aim of this study was to investigate the value of fractal dimension in separating normal and cancerous images, and to examine the relationship between fractal dimension and traditional texture analysis features. Forty-four normal images and 58 cancer images from sections of the colon were analyzed. A "leave one-out" analysis approach was used to classify the samples into each group. With fractal analysis there was a highly significant difference between groups (p < 0.0001). Correlation and entropy features showed greater differences between the groups (p < 0.0001). Nevertheless, the addition of fractal analysis to the feature analysis improved the sensitivity from 90% to 95% and specificity from 86% to 93%. PMID- 11936598 TI - Model-based processing scheme for quantitative 4-D cardiac MRI analysis. AB - In this paper, we present an integrated model-based processing scheme for cardiac magnetic resonance imaging (MRI), embedded in an interactive computing environment suitable for quantitative cardiac analysis, which provides a set of functions for the extraction, modeling, and visualization of cardiac shape and deformation. The methods apply four-dimensional (4-D) processing (three spatial and one temporal) to multiphase multislice MRI acquisitions and produce a continuous 4-D model of the myocardial surface deformation. The model is used to measure diagnostically useful parameters, such as wall motion, myocardial thicking, and myocardial mass measurements. The proposed model-based shape extraction method has the advantage of integrating local information into an overall representation and produces a robust description of cardiac cavities. A learning segmentation process that incorporates a generating-shrinking neural network is combined with a spatiotemporal parametric modeling method through functional basis decomposition. A multiscale approach is adopted, which uses at each step a coarse-scale model defined at the previous step in order to constrain the boundary detection. The representation accuracy starts from a coarse but smooth estimation of the approximate cardiac shape and is gradually increased to the desired detail. The main advantages of the proposed methods are efficiency, lack of uncertainty about convergence, and robustness to image artifacts. Experimental results obtained from application to clinical multislice multiphase MRI examinations of normal volunteers and patients with medical record of myocardial infarction were satisfactory in terms of accuracy and robustness. PMID- 11936599 TI - Quantitative comparison and analysis of brain image registration using frequency adaptive wavelet shrinkage. AB - In the field of template-based medical image analysis, image registration and normalization are frequently used to evaluate and interpret data in a standard template or reference atlas space. Despite the large number of image-registration (warping) techniques developed recently in the literature, only a few studies have been undertaken to numerically characterize and compare various alignment methods. In this paper, we introduce a new approach for analyzing image registration based on a selective-wavelet reconstruction technique using a frequency-adaptive wavelet shrinkage. We study four polynomial-based and two higher complexity nonaffine warping methods applied to groups of stereotaxic human brain structural (magnetic resonance imaging) and functional (positron emission tomography) data. Depending upon the aim of the image registration, we present several warp classification schemes. Our method uses a concise representation of the native and resliced (pre- and post-warp) data in compressed wavelet space to assess quality of registration. This technique is computationally inexpensive and utilizes the image compression, image enhancement, and denoising characteristics of the wavelet-based function representation, as well as the optimality properties of frequency-dependent wavelet shrinkage. PMID- 11936600 TI - Shape recovery algorithms using level sets in 2-D/3-D medical imagery: a state-of the-art review. AB - The class of geometric deformable models, also known as level sets, has brought tremendous impact to medical imagery due to its capability of topology preservation and fast shape recovery. In an effort to facilitate a clear and full understanding of these powerful state-of-the-art applied mathematical tools, this paper is an attempt to explore these geometric methods, their implementations and integration of regularizers to improve the robustness of these topologically independent propagating curves/surfaces. This paper first presents the origination of level sets, followed by the taxonomy of level sets. We then derive the fundamental equation of curve/surface evolution and zero-level curves/surfaces. The paper then focuses on the first core class of level sets, known as "level sets without regularizers." This class presents five prototypes: gradient, edge, area-minimization, curvature-dependent and application driven. The next section is devoted to second core class of level sets, known as "level sets with regularizers." In this class, we present four kinds: clustering-based, Bayesian bidirectional classifier-based, shape-based and coupled constrained based. An entire section is dedicated to optimization and quantification techniques for shape recovery when used in the level set framework. Finally, the paper concludes with 22 general merits and four demerits on level sets and the future of level sets in medical image segmentation. We present applications of level sets to complex shapes like the human cortex acquired via MRI for neurological image analysis. PMID- 11936601 TI - Medical image compression by sampling DCT coefficients. AB - Advanced medical imaging requires storage of large quantities of digitized clinical data. Due to the constrained bandwidth and storage capacity, however, a medical image must be compressed before transmission and storage. Among the existing compression schemes, transform coding is one of the most effective strategies. Image data in spatial domain will be transformed into spectral domain after the transformation to attain more compression gains. Based on the quantization strategy, coefficients of low amplitude in the transformed domain are discarded and significant coefficients are preserved to increase the compression ratio without inducing salient distortion. In this paper, we use an adaptive sampling algorithm by calculating the difference area between correct points and predicted points to decide the significant coefficients. Recording or transmitting the significant coefficients instead of the whole coefficients achieves the goal of compression. On the decoder side, a linear equation is employed to reconstruct the coefficients between two sequent significant coefficients. Simulations are carried out to different medical images, which include sonogram, angiogram, computed tomography, and X-ray images. Consequent images demonstrate the performance at compression ratios of 20-45 without perceptible alterations. In addition, two doctors are invited to verify that the decoded quality is acceptable for practical diagnosis. Therefore, our proposed method is found to preserve information fidelity while reducing the amount of data. PMID- 11936602 TI - An open architecture patient monitoring system using standard technologies. AB - Computer-aided bedside patient monitoring is applied in areas where real-time vital function analysis takes place. Modern bedside monitoring requires not only the networking of bedside monitors with a central monitor but also other standard communication interfaces. In this paper, a novel approach to patient monitoring is introduced. A patient monitoring system was developed and implemented based on an existing industry standard communication network, using standard hardware components and software technologies. The open architecture system design offers scalability, standard interfaces, and flexible signal interpretation possibilities. PMID- 11936603 TI - The distribution and history of nuclear weapons related contamination in sediments from the Ob River, Siberia as determined by isotopic ratios of plutonium and neptunium. AB - Isotopic ratios of Pu and Np measured in sediment cores from 5 locations in the Ob River drainage basin show clear evidence of input from sources other than global fallout (non-fallout sources). Historical contaminant records obtained by combining isotopic ratio information with chronological information indicate that non-fallout inputs are from several sources that have varied significantly over the past 50 years. Unique isotopic signatures observed in sediments from tributaries that drain areas containing known or suspected sources of non-fallout contamination are used to identify the source of materials in sediments collected at downstream locations. These data can lead to a better understanding of the transport behavior, fate, and relative importance of particle reactive, weapons related contaminants originating from the nuclear facilities Mayak. Tomsk-7, and Semipalitinsk, which lie within the drainage basin. From our work to date, we draw the following conclusions: (1) Persistent non-fallout contamination is observed in the Ob River above its confluence with the Irtysh River, indicating contamination from the Tomsk-7 facility. (2) Non-fallout contamination in the Tobol River above its confluence with the Irtysh River indicates contamination from the Mayak facility. (3) Non-fallout contamination in the Irtysh River above its confluence with the Tobol River indicates contamination from the Semipalitinsk weapons test site. (4) The occurrence of isotopic ratios in Ob Delta sediments that are similar to those observed in source tributaries suggests that contamination from at least two sources has been transported along the length of the river system. (5) Global fallout, a result of high-yield atmospheric weapons tests conducted by the FSU and USA primarily, is the dominant source of Pu and Np to the region; however, there have been brief periods when inputs from non-fallout sources exceeded those from global fallout. PMID- 11936604 TI - Aquatic mosses as a monitoring tool for 137Cs contamination in streams and rivers a field study from central southern Norway. AB - Mosses are frequently used as biomonitors for trace element pollution in the aquatic environment. The purpose of this study was to assess their usefulness as a tool in monitoring and in regional surveys of radioactive contamination. Specimens of the aquatic mosses, Fontinalis antipyretica and F. dalecarlica, were transplanted from non-contaminated areas to streams and rivers in the Norwegian Jotunheimen Mountains and neighbouring lowland areas that had received radioactive fallout after the Chernobyl accident. Equilibrium concentrations of 137Cs in the exposed mosses were reached after a few weeks. Two series from 20 streams in 1994 and 1996 show linear correlations between activities in water and moss samples and biomagnification ratios of 10(4) - 10(5). We conclude that mosses are better suited for monitoring purposes than water samples, because they provide values integrated over weeks while the radioactivity in surface waters can be subject to rapid variations according to hydrological events. The activity concentrations in aquatic mosses can be easily measured with good precision even when aqueous concentrations are below the limit of detection. Use of aquatic mosses also reduces the logistic problems of transporting large volumes of water, especially in areas inaccessible by road. PMID- 11936605 TI - An analysis of the environmental mobility of radiostrontium from weapons testing and Chernobyl in Finnish river catchments. AB - The mobility of radiostrontium within the Arctic environment and surrounding area has been studied by analysing the mobility of 90Sr in river catchments that are within Finland. The environmental mobility of 90Sr deposited by both nuclear weapons testing and the Chernobyl accident has been investigated in five Finnish river catchments. Different models assessing the time-dependent mobility of 90Sr have been evaluated. No significant differences were found between the mobility of 90Sr from nuclear weapons tests and from the Chernobyl accident. Model parameters obtained by fitting to the measurements of the deposition and runoff rates of the nuclear weapons test fallout gave predictions which were consistent with the mid- and long-term contamination by the Chernobyl fallout. A comparison of 90Sr with 137Cs showed that they had similar mobility on deposition but, as time passed, the relative mobility of 90Sr increased with respect to 137Cs over a period of 5-8 years. Once the relative migration of 90Sr with respect to 137Cs reached equilibrium, its runoff rate was, on average, approximately an order of magnitude greater than 137Cs. PMID- 11936606 TI - An assessment of the reported leakage of anthropogenic radionuclides from the underground nuclear test sites at Amchitka Island, Alaska, USA to the surface environment. AB - Three underground nuclear tests representing approximately 15-16% of the total effective energy released during the United States underground nuclear testing program from 1951 to 1992 were conducted at Amchitka Island, Alaska. In 1996, Greenpeace reported that leakage of radionuclides, 241Am and 239+240Pu, from these underground tests to the terrestrial and freshwater environments had been detected. In response to this report, a federal, state, tribal and non governmental team conducted a terrestrial and freshwater radiological sampling program in 1997. Additional radiological sampling was conducted in 1998. An assessment of the reported leakage to the freshwater environment was evaluated by assessing 3H values in surface waters and 240Pu/239Pu ratios in various sample media. Tritium values ranged from 0.41 Bq/l +/- 0.11 two sigma to 0.74 Bq/1 +/- 0.126 two sigma at the surface water sites sampled, including the reported leakage sites. Only at the Long Shot test site, where leakage of radioactive gases to the near-surface occurred in 1965. were higher 3H levels of 5.8 Bq/1 +/- 0.19 two sigma still observed in 1997, in mud pit #3. The mean 240Pu/239Pu for all of the Amchitka samples was 0.1991 +/- 0.0149 one standard deviation, with values ranging from 0.1824 +/- 1.43% one sigma to 0.2431 +/- 6.56% one sigma. The measured 3H levels and 240Pu/239Pu ratios in freshwater moss and sediments at Amchitka provide no evidence of leakage occurring at the sites reported by Buske and Miller (1998 Nuclear-Weapons-Free America and Alaska Community Action on Toxics, Anchorage, Ak, p.38) and Miller and Buske (1996 Nuclear Flashback: The Return to Anchitka, p.35). It was noted that the marine sample; 240Pu/239Pu ratios are statistically different than the global fallout ratios presented by Krey et al. (1976) and Kelley, Bond, and Beasley (1999). The additional non fallout component 240Pu/239Pu ratio, assuming a single unique source, necessary to modify the global fallout 240Pu/239Pu ratio to that measured in the marine samples is on the order of 0.65 (Hameedi, Efurd, Harmon, Valette-Silver, & Robertson, 1999; Kelley et al., 1999). While this potentially suggests another plutonium source, such as high burn-up nuclear reactor fuel, rather than underground nuclear tests, the uncertainties in analyses and environmental processes need to be fully assessed before any conclusion can be reached. Further work is needed to evaluate these findings and to support any radiological assessment of the marine environment surrounding Amchitka. Based on geohydrological testing and modeling, leakage from the Amchitka Underground Nuclear Tests is projected to occur to the marine environment (Claassen, 1978; Fenske, 1972; Wheatcraft, 1995). PMID- 11936607 TI - Radioactive fallout in Norway from atmospheric nuclear weapons tests. AB - Historical data on radioactivity in air and precipitation samples have been collected and analysed from study sites in Norway. The purpose of the study was to investigate the correlation between air concentration, precipitation and deposition, and identify areas with high deposition. Areas with high precipitation have been compared with monitoring stations in other countries. The base line data contain measurements of total beta in air and precipitation on a daily basis for the period 1956-1982. Radioactive fallout correlated strongly with annual precipitation which varies from 280 to 4200mm per year in Norway. The deposition of 137Cs was calculated to be 3.23+/-1.20kBq/m2 per 1,000 mm precipitation for the period 1955-1975. Also, the relationship between total beta and 137Cs has been investigated, in order to estimate the age of fallout. The age of fallout in Norway ranges from 3 to 9 months during the test periods, which is considerably shorter than the global average, where the mean residence time for debris in the lower stratosphere is estimated to be 1.3 years. There is no evidence of local fallout from tests on Novaya Zemlya reaching Norwegian areas. PMID- 11936608 TI - Estimation of critical loads for radiocaesium in Fennoscandia and Northwest Russia. AB - The application of the critical loads methodology for radioactive contamination of Arctic and sub-arctic ecosystems, where natural and semi-natural food products are important components of the diet of many people, is proposed and discussed. The critical load is herein defined as the amount of radionuclide deposition necessary to produce radionuclide activity concentrations in food products exceeding intervention limits. The high transfer of radiocaesium to reindeer meat gives this product the lowest critical load, even though the intervention limit is relatively high compared with other products. Ecological half-lives of radiocaesium in natural and semi-natural products are often very long, and it is therefore important to take account of contamination already present in the event of an accident affecting areas where such products are important. In particular, the long ecological half-life for radiocaesium in moose meat means that the critical load is highly sensitive to prior deposition. An example of the potential application of the method for emergency preparedness is given for the Chernobyl accident. PMID- 11936609 TI - Residual radioactive contamination at the peaceful underground nuclear explosion sites "Craton-3" and "Crystal" in the Republic of Sakha (Yakutia). AB - In this paper, results are reported on the concentrations and activity ratios of 137Cs, 239+240Pu and 238Pu in soils, lichen and bottom sediments collected at the peaceful underground nuclear explosion sites "Craton-3" and "Crystal" (Republic of Sakha (Yakutia), North-East Siberia). At the "Craton-3" site, 239- 240Pu concentrations in the range 6.2 mBq/g to 5.9 Bq/g in surface soils (0-5 cm) and 239+240Pu concentrations up to 7.4 Bq/g in lichen were observed. The 137Cs concentration in measured soils range from 0.52 to 216 Bq/g for the near-plume and from 1.65 to 21.5 Bq/g for the far-plume. In lichen, 137Cs concentration varied from a global background level up to 251 Bq/g. Radioactive contamination at "Crystal" demonstrates an extremely irregular distribution of 239,240Pu and 137Cs in environmental samples taken at locations within 150 m radius of the site. Further work on compiling detailed maps of radioactive contamination of the territory around "Craton-3" and "Crystal" is discussed. PMID- 11936610 TI - Atmospheric transport patterns and possible consequences for the European North after a nuclear accident. AB - The main purpose of this study is to examine possible impacts and consequences of a hypothetical accident at the Kola nuclear plant in north-west Russia on different geographical regions: Scandinavia, central Europe, European FSU and Taymyr. The period studied is 1991-1996. An isentropic trajectory model has been used to calculate forward trajectories that originated over the nuclear accident region. Atmospheric transport patterns were identified using the isentropic trajectories and a cluster analysis technique. From the trajectory model results, a number of cases were chosen for examination in detail using more complete transport models. For this purpose, the models MATHEW/ADPIC, DERMA and a newly developed FOA Random Displacement Model have been used to simulate the radionuclide transport and contamination in the case of a nuclear accident and their results have been compared with those of the trajectory modelling. Estimation of the long-term consequences for populations after an accident has been performed for several specific dates by empirical models and correlation between fallout and doses to humans on the basis of the Chernobyl accident exposures in Scandinavia. PMID- 11936611 TI - Assessment of current exposure levels in different population groups of the Kola Peninsula. AB - Activity concentrations of 137Cs and 90Sr in samples of vegetation and natural food products collected in the Kola Peninsula in 1998 and 1999 indicate a very slow decrease in contamination levels during the last decade, mainly due to the physical decay of the radionuclides. The activity concentrations of 137Cs in reindeer meat decreased with a half-life of about 9 years. 137Cs in lichen, moss and fungi is significantly higher than in natural vegetation (grasses) and agricultural plants (potatoes). The activity concentrations of 137Cs in reindeer meat were two orders of magnitude higher than those in locally produced beef and pork. Consumption of reindeer meat, fish, mushrooms and berries constituted the main contribution to the internal dose from 137Cs and 90Sr for reindeer-breeders in the Lovozero area. The estimated committed doses due to 137Cs intake in this group were about 10 microSv per month in summer 1998 and 15 microSv per month in winter, 1999. There was good agreement between internal dose estimates based on intake assessment and whole body measurements. The population of Umba settlement, which is not involved in reindeer breeding, received individual committed doses due to 137Cs intake of about 0.5 microSv per month, about a factor of 20 less than the reindeer-breeders in Lovozero. In this case, the main contribution to the internal dose of the general population came from consumption the of 137Cs in mushrooms and forest berries. The contribution of 90Sr to the internal dose varied from 1% to 5% in the different population groups studied. PMID- 11936612 TI - Temporal trends for 99Tc in Norwegian coastal environments and spatial distribution in the Barents Sea. AB - The objective of this study was to reassess 99Tc transit times and transfer factors, from Sellafield to northern Norway, and to determine the extent of 99Tc migration to the Barents Sea. Filtered seawater samples were collected on a monthly basis from Hillesoy, northern Norway, and in February 1999 from the Barents Sea. Results showed an increase in levels of 99Tc at Hillesoy where activity concentrations have increased from a baseline of 0.2-0.4Bq m(-3) to a maximum of 1.6 Bq m(-3). A transit time of 42 months and a transfer factor of 6Bq m(-3) per PBq a(-1) have been derived, using cross-correlation analysis. The current study predicts that future levels are unlikely to increase dramatically over the levels observed in 1998. Levels of 99Tc in the Barents Sea ranged from 0.2 Bq m(-3) to 1.1 Bq m(-3) showing the influence of new 99Tc inputs by early 1999. PMID- 11936613 TI - Radioactive contamination in the Arctic--sources, dose assessment and potential risks. AB - Arctic residents, whose diets comprise a large proportion of traditional terrestrial and freshwater foodstuffs, have received the highest radiation exposures to artificial radionuclides in the Arctic. Doses to members of both the average population and selected indigenous population groups in the Arctic depend on the rates of consumption of locally-derived terrestrial and freshwater foodstuffs, including reindeer/caribou meat, freshwater fish, goat cheese, berries, mushrooms and lamb. The vulnerability of arctic populations, especially indigenous peoples, to radiocaesium deposition is much greater than for temperate populations due to the importance of terrestrial, semi-natural exposure pathways where there is high radiocaesium transfer and a long ecological half-life for this radionuclide. In contrast, arctic residents with diets largely comprising marine foodstuffs have received comparatively low radiation exposures because of the lower levels of contamination of marine organisms. Using arctic-specific information, the predicted collective dose is five times higher than that estimated by UNSCEAR for temperate areas. The greatest threats to human health and the environment posed by human and industrial activities in the Arctic are associated with the potential for accidents in the civilian and military nuclear sectors. Of most concern are the consequences of potential accidents in nuclear power plant reactors, during the handling and storage of nuclear weapons, in the decommissioning of nuclear submarines and in the disposal of spent nuclear fuel from vessels. It is important to foster a close association between risk assessment and practical programmes for the purposes of improving monitoring, formulating response strategies and implementing action plans. PMID- 11936614 TI - 129I/127I, 129I/137Cs and 129I/99Tc in the Norwegian coastal current from 1980 to 1998. AB - Discharges of the nuclides 129I, 137Cs and 99Tc from the nuclear reprocessing facilities at Sellafield (UK) and La Hague (France) are very useful as oceanic tracers. On the basis of 129I/127I, 137Cs and 99Tc measurements in archived seaweeds, the ratios of 129I/127I, 129I/137Cs and 129I/99Tc have been estimated in seawater at two locations (Utsira and Kiberg) in the Norwegian Coastal Current from 1980 to 1998. These ratios, which vary up to two orders of magnitude over this period, are potentially very interesting for determining "transit times" in the Arctic and North Atlantic oceans. While the long-term trends in these ratios are quite clear, measurements in monthly and bimonthly samples show considerable structure. Further studies are required to determine the exact origin of this structure, which may be a limiting factor in the time resolution that can be obtained with these parameters. PMID- 11936615 TI - Transport of plutonium in surface and sub-surface waters from the Arctic shelf to the North Pole via the Lomonosov Ridge. AB - New data on the levels and long-range transport of plutonium in the Arctic Ocean, recorded in the course of two expeditions to this zone in 1994 and 1996, are discussed in this paper. Specifically, approximately 100 plutonium measurements in surface and sub-surface water sampled at 58 separate stations throughout the Kara, Laptev and East Siberian Seas, as well as along latitudinal transects across the Lomonosov Ridge, are reported and interpreted in terms of the circulation pathways responsible for the transport of this element from the North Atlantic to the Arctic Shelf and into the Arctic interior. In addition, the behaviour of plutonium in its transit through the vast Arctic shelf seas to open waters under extreme environmental conditions is discussed in terms of the partitioning of plutonium between filtered (<0.45 microm) seawater and suspended particulate, and its association with colloidal matter. Finally, limited evidence of the presence of a colloidal plutonium component in Arctic waters subject to direct riverine input is adduced. PMID- 11936616 TI - Modified approach to modelling radiological consequences from releases into the marine environment. AB - One of the general assumptions for box modelling of the dispersion of radionuclides in marine systems relates to instantaneous mixing in each box which, in turn, results in practical calculations involving instantaneous mixing in the whole of oceanic space. A new approach to box modelling, which includes dispersion of radionuclides as a function of time, was therefore developed in order to provide a better and more realistic/physical approximation to reality relative to traditional box modelling. The novel and significant practical features of the approach are discussed. Calculations of concentrations of radionuclides in the marine environment and doses to man for some scenarios indicate differences of up to orders of magnitude between the traditional and new approaches to box modelling. PMID- 11936617 TI - Colon cancer prevention with NO-releasing NSAIDs. AB - A seminal advance in the prevention of colon cancer has been the observation that nonsteroidal antiinflammatory drugs (NSAIDs) reduce the incidence of and mortality from colon cancer by about half. Among current efforts to overcome the side effects of NSAIDs, an important limitation for their application as chemopreventive agents, is the synthesis of nitric oxide-releasing NSAIDs. These novel compounds may display greater safety and greater efficacy compared to their parent traditional NSAIDs and thus hold significant promise as chemopreventive agents against human colon cancer. In this review we discuss salient features of their pharmacology, in vitro and animal data pertaining to colon cancer, their mechanisms of action, and assess their potential in the chemoprevention of colon cancer. PMID- 11936618 TI - The tissue-specific, compensatory expression of cyclooxygenase-1 and -2 in transgenic mice. AB - Prostaglandins are essential regulators of tissue homeostasis, reproduction and inflammation. We have recently shown that cells derived from cyclooxygenase (COX) deficient mice express higher, compensatory levels of the remaining COX isozyme [Kirtikara et al., J. Exp. Med., 187, 517 (1998)]. To assess this compensatory expression phenomenon in vivo, we quantified COX-1 and COX-2 mRNA levels in various organs of COX-1- and COX-2-ablated mice using a reverse transcriptase polymerase chain reaction (RT-PCR) method. We found that COX-1 and COX-2 mRNAs in the brains of COX-ablated mice were elevated > 2-fold compared with wild-type (WT) animals. COX-2 mRNA was enhanced approximately 2-fold in the kidneys and stomachs of COX-1-deficient mice while COX-1 expression remained unchanged. Conversely, the livers of COX-2-deficient mice expressed 15-fold higher COX-1 mRNA levels, while hepatic COX-2 mRNA levels were not significantly altered in the COX-1-ablated mice. Steady state levels of COX-1 and COX-2 mRNAs in the hearts, lungs and spleens of WT, COX-1- and COX-2-deficient mice were indistinguishable from each other. Peritoneal macrophages isolated from COX-1- and COX-2-ablated mice also expressed significantly higher steady-state levels of cytoplasmic phospholipase A2 and 5-lipooxygenase mRNAs suggesting a global upregulation of eicosanoid biosynthetic pathways in COX-deficient mice. These data suggest that expression of both COX-1 and COX-2 can be re-programmed to compensate for the lack of both alleles of the alternate COX gene in transgenic mice. PMID- 11936619 TI - Dietary fat and salivary prostaglandin E2. AB - A study of the relation between dietary fat intake and salivary prostaglandin E2 was undertaken in Transkei, South Africa. Samples of saliva were obtained from (1) Transkeians on a very low fat diet, (2) Transkeians on a low fat diet, (3) Transkeians on a medium fat diet, (4) British patients. Salivary PGE2 means for the groups were (1) 2357 pg/ml, (2) 2020 pg/ml, (3) 733 pg/ml, (4) 312.5 pg/ml. Differences between groups 1 and 3, 1 and 4, and 2 and 4 were significant. Rural Transkeians on a low-fat diet have an elevated level of PGE2 in saliva. As fat increases in the diet, PGE2 in saliva tends towards the level found in those who eat a western diet. An increase in the level of PGE2 production in the upper gastrointestinal tract and in the tissues of the body may be a factor in promoting cancer of the esophagus and diseases favored by Th1 immune dysfunction. PMID- 11936621 TI - Lessons learnt at ICA2001. IWA Conference on Instrumentation, Control and Automation. AB - This paper gives an introduction as well as some general experiences and conclusions from the 1st IWA Conference on Instrumentation, Control and Automation (ICA2001), June 3-7, 2001, Malmo, Sweden. The highlights from the different sessions of the conference are summarised and some challenges for the coming years, with regard to ICA within the field of wastewater treatment, are also discussed. PMID- 11936620 TI - Selective inhibition of COX-2 is beneficial to mice infected intranasally with VSV. AB - Cyclooxygenase (COX) is the key enzyme for prostaglandin (PG) synthesis. PGs are mediators of many critical physiological and inflammatory responses. There are two isoforms, COX-1 and COX-2, both of which are constitutively expressed in the central nervous system (CNS). Studies have shown that COX-1 and COX-2 are involved in physiological and pathological conditions of the brain. However, little is known about the role(s) of COX in the host defense system against a viral infection in the CNS. In this report, we used Vesicular Stomatitis Virus (VSV) induced acute encephalitis to distinguish between the contribution(s) of the two isoforms. COX-2 activity was inhibited with a COX-2 selective drug, celecoxib (Celebrex), and COX-1 was antagonized with SC560. We found that inhibition of COX-2 led to decreased viral titers, while COX-1 antagonism did not have the same effect at day 1 post infection. 5-lipooxygenase (5-LO) expression and neutrophil recruitment in the CNS were increased in celecoxib-inhibited mice. Furthermore, mice treated with celecoxib expressed more Nitric Oxide Synthase-1 (NOS-1), a crucial component of the innate immune system in the restriction of VSV propagation. The expression of type 1 cytokines, IFN-gamma and IL-12, were also increased in celecoxib-treated mice. PMID- 11936623 TI - On the use of fluorescence measurements to characterize wastewater. AB - This paper examines the correlations between some water quality parameters and fluorescence intensities and spectra in filtered wastewater using a 280 nm excitation wavelength. We did not obtain satisfying linear relationships between fluorescence and COD or TOC (r2 approximately 0.4) for any of the emission wavelengths used between 320 and 400 nm (especially at 355 nm, the average emission lambda(max.)). The relationships with NK were better (r2 approximately 0.7); leading us to suggest that one evaluates mainly the organic nitrogen content at lambda(ex.) = 280 nm. Whole spectra processing did not improve the correlations. Gel permeation chromatography yielded markedly different TOC and fluorescence chromatograms, which explains the difficulty of correlating organic content and fluorescence. Other investigations are necessary before spectrofluorimetry can be used as a reliable technique for on-line wastewater pollution estimation. PMID- 11936622 TI - Soft sensors for control of nitrogen and phosphorus removal from wastewaters by neural networks. AB - In this paper, we describe the results of research aimed to evaluate the possibility of using a neural network (NN) model for predicting biological nitrogen and phosphorus removal processes in activated sludge, utilising oxidation reduction potential (ORP) and pH as NN inputs. Based on N and P concentrations predictions obtained via the NN, a strategy for controlling sequencing batch reactors (SBRs) phases duration, optimising pollutants removal and saving energy, is proposed. The NN model allowed us to reproduce the concentration trends (change in slope, or process end), with satisfactory accuracy. The NN results were generally in good agreement with the experimental data. These results demonstrated that NN models can be used as "soft on-line sensors" for controlling biological processes in SBRs. By monitoring ORP and pH, it is possible to recognise the N and P concentrations during different SBRs phases and, consequently, to identify the end of the biological nutrient removal processes. This information can then be used to design control systems. PMID- 11936624 TI - Robustness and economic measures as control benchmark performance criteria. AB - The objective of this paper was to investigate the usefulness of new multi criteria measures to evaluate a control strategy through the Benchmark protocol. Using a case study in which respirometry-based control strategies are evaluated, the proposed measures were calculated. An economic index including weighted investment and operating costs (termed Total Cost Index--TCI) appears more powerful than a grey-scale presentation approach. Using the latter approach, it is hard to reflect the relative importance of the criteria investigated, which makes practical decisions rather difficult. In addition, a Robustness Index (RI) is proposed that allows us to evaluate the transferability of control strategies to situations different from the ones defined in the benchmark protocol. Finally, the case study shows that it may be advisable to replace the currently used open loop benchmark reference by a plant in which dissolved oxygen is controlled in all aerated reactors. This quite simple strategy also turned out to be the best one among all evaluated strategies. PMID- 11936625 TI - Wastewater treatment benchmark: what can be achieved with simple control? AB - In this paper a simple control strategy is applied to and assessed on the wastewater treatment benchmark. The controllers used in the presented control strategy are PI controllers, feedforward control and a step-feed procedure. The controlled variables are not directly the effluent concentrations but other process variables which have an effect on the effluent. The setting of set-points is also analyzed to select the values with the best performance. Set-point analysis has shown that with an optimal setting of set-points under stormy influent conditions, the achieved plant performance is also retained for rainy and dry influent conditions. The evaluation of plant performance indicates that with the proposed control strategy, a lower number of effluent violations has been achieved, as well as lower energy consumption and lower sludge production, when compared to results published up to now. Only the effluent quality criterion deteriorated. PMID- 11936626 TI - Control of an activated sludge process with nitrogen removal--a benchmark study. AB - In this paper, a simulation benchmark of a predenitrifying activated sludge process is used to evaluate a number of control strategies. A main procedure has been to use feedforward terms that are based on simplified physical models. Important mass balance relations may then be incorporated in the control law. The nitrate level in the last anoxic zone is controlled by the dosage of an external carbon source and the nitrate level in the last aerobic zone is controlled by the internal recirculation flow rate. The ammonia level is controlled by a DO set point controller. In order to be able to have as high a sludge level as possible without sludge escape, the sludge blanket height in the settler is controlled by the excess sludge flow rate. Compared to the default set up of the benchmark, the controllers could reduce the effluent nitrate significantly whereas the effluent ammonia was only marginally decreased. The main problem is that the aeration capacity defined in the benchmark is too low. PMID- 11936627 TI - Evaluation of several respirometry-based activated sludge toxicity control strategies. AB - Four different strategies including influent storage and reintroduction, step feeding, rapid sludge recycle and waste sludge storage were evaluated using the denitrification layout of the IWA simulation benchmark. The control objective was to minimise deterioration in effluent quality caused by a certain toxic input event. In these strategies the maximum specific respiration rate (Rmax) was selected as a measured and controlled variable. To simplify the analysis, the toxicant was assumed to be a soluble and nonbiodegradable substance. Two toxic influent files were developed with square-wave input lasting 3 hours. To detect the influent toxicity, a pseudo-online flow-through respirometer was applied. A number of simulations were performed and the results suggested that the influent storage and reintroduction strategy provided the most optimistic results and other strategies could not mitigate the toxic effect. The influent storage and reintroduction strategy strongly depended on reintroduction flow rate from the storage tank. The simulation according to reintroduction flow could estimate the time required for completely treating toxic wastewater stored in the storage tank. Also the IWA simulation benchmark was enhanced to evaluate toxicity effect on the activated sludge process. PMID- 11936628 TI - Evaluation of control strategies using an oxidation ditch benchmark. AB - This paper presents validation and implementation results of a benchmark developed for a specific full-scale oxidation ditch wastewater treatment plant. A benchmark is a standard simulation procedure that can be used as a tool in evaluating various control strategies proposed for wastewater treatment plants. It is based on model and performance criteria development. Testing of this benchmark, by comparing benchmark predictions to real measurements of the electrical energy consumptions and amounts of disposed sludge for a specific oxidation ditch WWTP, has shown that it can (reasonably) be used for evaluating the performance of this WWTP. Subsequently, the validated benchmark was then used in evaluating some basic and advanced control strategies. Some of the interesting results obtained are the following: (i) influent flow splitting ratio, between the first and the fourth aerated compartments of the ditch, has no significant effect on the TN concentrations in the effluent, and (ii) for evaluation of long term control strategies, future benchmarks need to be able to assess settlers' performance. PMID- 11936629 TI - Implementation of storage tanks on the COST 624 benchmark. AB - To test the improvement that can be expected in terms of effluent quality of an wastewater treatment plant by activated sludge, an equalisation tank and a storm tank, designed to damp the influent variations under different weather conditions, have been implemented in front of a benchmark plant used to evaluate control strategies. The equalisation tank improves significantly the effluent quality in any weather condition but at a high operation cost due to extra pumping, while the storm tank without by-pass improves the effluent quality in rainy periods with a small increase in cost operation over the no tank case. PMID- 11936630 TI - Sensor system using polarization analysis method to monitor oil-on-water in water purification plants and rivers. AB - Some 80% of accidental pollution in river water is caused by oil spills. Oil spills can cause serious damage such as suspension of water intake at water purification plants and major harm to ecosystems in the lower reaches of rivers. This is because oil-on-water tends to spread easily, quickly exacerbating the damage. To address this problem, an automated, continuous sensor system with high sensitivity can be used for early detection of spill accidents. We have developed a sensor system for detecting oil-on-water based on a polarization analysis method. Its advantages include: a) no direct contact with sample water; b) minimal maintenance; c) largely unaffected by foreign matter and waves on the water surface; and d) much higher sensitivity than simple visual observation. This paper describes the measurement principle and configuration of the sensor system, and discusses the results of sensitivity tests and tests on the influence of water turbidity, foreign matter and waves. We will also consider some of the limitations of the new system. PMID- 11936631 TI - Water monitoring system for oil contamination using polymer-coated quartz crystal microbalance chemical sensor. AB - A water monitoring system with a new chemical sensor for oil contamination was developed. The sensor had an organic polymer film on a quartz crystal microbalance (QCM). The organic film was a hydrocarbon polymer and had high affinities for the organic compounds of petroleum products such as gasoline, kerosene, diesel oil and fuel oil. The monitoring system as composed of a sampling part, a purging part, a humidity control part and a sensor part. The oil in contaminated river water could be detected whose threshold odour number (TON) was less than three. The detecting time was less than 5 min depending on the oil kindness. This system was tested using artificially contaminated river water with the oils, to be found that the sensitivity was kept steady for longer than 6 months with 400 detections of diesel oil and heavy oil. Moreover, the oil kind could be discriminated with only one sensor device by analyzing the desorption response curves obtained by flowing a clean air on the sensor instead of the purging air. PMID- 11936632 TI - Anoxic activated sludge monitoring with combined nitrate and titrimetric measurements. AB - An experimental procedure for anoxic activated sludge monitoring with combined nitrate and titrimetric measurements is proposed and evaluated successfully with two known carbon sources, acetate and dextrose. For nitrate measurements an ion selective nitrate electrode is applied to allow for frequent measurements, and thereby the possibility for detailed determination of the denitrification biokinetics. An internal nitrate electrode calibration is implemented in the experiments to avoid the often-encountered electrode drift problem. It was observed that the best experimental design was with the carbon source in excess, since excess nitrate provoked nitrite build-up thereby complicating the data interpretation. A conceptual model could quantitatively describe the experimental observations and thus link the experimentally measured proton production with the consumption of electron acceptor and carbon source during denitrification. PMID- 11936633 TI - In-line fast OUR (oxygen uptake rate) measurements for monitoring and control of WWTP. AB - This paper shows how using in-reactor on-line monitoring of OUR in the aerobic reactors of an A2/O (anaerobic, anoxic, oxic) nutrient removal system, the implemented control system is able to improve the nitrogen removal of the plant. The control system adapts the operation to different load conditions, without the need for additional information provided by other analysers. In particular, these OUR measurements and on-line information available in the distributed control system (temperature, biomass concentration and flows) are used for the estimation of the COD load in the influent, and automatically applied in the adaptation of the operational mode of the nutrient removal process. The estimation is realised using Artificial Neural Networks models trained with available data from the system. The operational changes consist in the modification of the total aerated volume in the pilot plant as a function of the load estimation. With the implemented control system based on COD load estimation, an increase in nitrogen removal of 10% was obtained with respect to a system working with fixed aeration volume, and using a suitable amount of energy in each case. PMID- 11936634 TI - Detection of Escherichia coli in water by culture-based amperometric and luminometric methods. AB - The application of amperometric biosensor- and chemiluminiscence based methods for rapid detection of viable E. coli in water has been investigated. An amplification of the amperometric signal by a factor of 4 was obtained when the cellobiose dehydrogenase (CDH) biosensor was used instead of a plain graphite electrode for detection of b-galactosidase (b-GAL) activity at 22.5 degrees C. A linear correlation was demonstrated for detection time (DT) vs. initial concentrations (logarithmic units) of E. coli IT1 and E. coli in environmental samples, respectively, by use of the CDH biosensor or a chemiluminometric technique. The study has shown that an E. coli concentration > or = 10(4) cfu/100 mL in environmental samples was determined by the CDH biosensor within one working day. However, further reduction of the DT can be obtained, e.g. by increasing the signal amplification factor using other biosensors. PMID- 11936635 TI - Detecting odorous materials in water using quartz crystal microbalance sensors. AB - Water drawn from rivers into purification plants must be checked for the presence of odorous materials because oil or organic-solvent contamination of the water may occur. If the detection of odorous materials in water is untimely or fails, the consequences can be serious. Therefore, the water must be checked continuously. We have developed a water-monitoring system that uses a highly sensitive electronic nose consisting of quartz crystal microbalance sensors to detect odorous materials in water. The nose is sensitive enough to detect petroleum hydrocarbons without water vapor at a low-ppb level. However, the nose is very sensitive to humidity and temperature. We have thus developed a method for accurately maintaining the humidity and temperature in the sensor cell. Experimental results show that the developed system can quickly detect contaminated water that was mixed with gasoline, kerosene, or benzene (concentration: several hundred ppb level), and we should be able to classify the pollutant by using pattern recognition of the dynamic sensor response. PMID- 11936636 TI - Low-cost failure sensor design and development for water pipeline distribution systems. AB - This paper describes the design and development of a new sensor which is low cost to manufacture and install and is reliable in operation with sufficient accuracy, resolution and repeatability for use in newly developed systems for pipeline monitoring and leakage detection. To provide an appropriate signal, the concept of a "failure" sensor is introduced, in which the output is not necessarily proportional to the input, but is unmistakably affected when an unusual event occurs. The design of this failure sensor is based on the water opacity which can be indicative of an unusual event in a water distribution network. The laboratory work and field trials necessary to design and prove out this type of failure sensor are described here. It is concluded that a low-cost failure sensor of this type has good potential for use in a comprehensive water monitoring and management system based on Artificial Neural Networks (ANN). PMID- 11936637 TI - Disturbance detection and isolation in the activated sludge process. AB - This paper proposes a new fault detection and isolation (FDI) method. This method monitors the distribution of process data and detects changes in this distribution, which reflect changes in the corresponding operating condition. A modified dissimilarity index and a FDI technique are defined to quantitatively evaluate the difference between data sets. This technique considers the importance of each transformed variable in the multivariate system. The FDI technique is applied to a benchmark simulation and to data from a real wastewater treatment plant. Simulation results show that it immediately detects disturbances and automatically distinguishes between serious and minor anomalies for various types of fault. The method not only detects the disturbances, but also isolates the scale of the disturbance, facilitating the interpretation of the disturbance source. The proposed monitoring technique is found to be appropriate for analyzing the biological wastewater treatment process, which is characterized by a variety of fault and disturbance sources and non-stationary characteristics. PMID- 11936638 TI - Adaptive multiscale principal components analysis for online monitoring of wastewater treatment. AB - Fault detection and isolation (FDI) are important steps in the monitoring and supervision of industrial processes. Biological wastewater treatment (WWT) plants are difficult to model, and hence to monitor, because of the complexity of the biological reactions and because plant influent and disturbances are highly variable and/or unmeasured. Multivariate statistical models have been developed for a wide variety of situations over the past few decades, proving successful in many applications. In this paper we develop a new monitoring algorithm based on Principal Components Analysis (PCA). It can be seen equivalently as making Multiscale PCA (MSPCA) adaptive, or as a multiscale decomposition of adaptive PCA. Adaptive Multiscale PCA (AdMSPCA) exploits the changing multivariate relationships between variables at different time-scales. Adaptation of scale PCA models over time permits them to follow the evolution of the process, inputs or disturbances. Performance of AdMSPCA and adaptive PCA on a real WWT data set compared and contrasted. The most significant difference observed was the ability of AdMSPCA to adapt to a much wider range of changes. This was mainly due to the flexibility afforded by allowing each scale model to adapt whenever it did not signal an abnormal event at that scale. Relative detection speeds were examined only summarily, but seemed to depend on the characteristics of the faults/disturbances. The results of the algorithms were similar for sudden changes, but AdMSPCA appeared more sensitive to slower changes. PMID- 11936639 TI - A neural network approach to burst detection. AB - This paper describes how hydraulic and water quality data from a distribution network may be used to provide a more efficient leakage management capability for the water industry. The research presented concerns the application of artificial neural networks to the issue of detection and location of leakage in treated water distribution systems. An architecture for an Artificial Neural Network (ANN) based system is outlined. The neural network uses time series data produced by sensors to directly construct an empirical model for predication and classification of leaks. Results are presented using data from an experimental site in Yorkshire Water's Keighley distribution system. PMID- 11936640 TI - Dinitrogen oxide detection for process failure early warning systems. AB - A number of experiments were conducted in order to establish whether the concentration of N2O in the off-gas from an activated sludge pilot plant could be used as a indicator for monitoring the nitrification process and as an early indication of ammonia appearing in the plant effluent. A strong correlation was found between ammonia shock loads and the concentration of N20 in the off-gas from the aeration tank for ammonia shock loads and dissolved oxygen depletion. When subjecting the experimental setup to doses of a nitrification inhibitor (allylthiourea) a similar pattern was seen with a correlation between nitrite build up in the aeration tank and concentration increase of N2O in the off-gas from the aeration tank. The results from this work suggest the concentration and the changes in the concentration of N2O in the exhaust gas from a nitrifying process may be a useful parameter for monitoring nitrifying activated sludge processes. PMID- 11936642 TI - A method for automatic validation of long time series of data in urban hydrology. AB - Modelling in urban hydrology is largely based on the analysis of long time series of data. The quality of the results strongly depends on the quality of the data used. Doubtful or wrong data must be detected and eventually substituted by reliable ones when it is feasible before any further exploitation. This paper deals with the development of an automatic pre-validation procedure that detects doubtful and not reliable data, in order to facilitate their interpretation. This procedure consists in applying a set of seven tests based on the following criteria: the functioning state of the sensor, the physical range of the quantity, the locally realistic range, the duration since the last maintenance of the sensor, the signal's gradient, material redundancy and analytical redundancy. The results of the tests are coded with the letter A for reliable values, B for doubtful values and C for wrong values. After this automatic prevalidation, the ultimate validation of values marked B and C is carried out manually by the operator, with the assistance of specifically developed visual and graphical tools. PMID- 11936641 TI - Model based prediction of the clogging of an anaerobic fixed bed reactor. AB - This paper presents the use of nonlinear constrained optimization techniques in order to detect and evaluate the degree of clogging in an anaerobic fixed bed reactor. First, experimental results show that the validity of a mass balance model can degrade over the time. Using the available model of the process and nonlinear constrained optimization tools, it is established that these changes can be due to the decrease of the liquid volume into the reactor while the mean values of biomass concentrations increase, leading to the clogging of the reactor. These theoretical results are confirmed experimentally in evaluating the hydraulic retention time of the reactor using a tracer. PMID- 11936643 TI - Application of toxicity monitor using nitrifying bacteria biosensor to sewerage systems. AB - Toxic substances may be included in wastewater influent and can damage biological processing of wastewater treatment, therefore continuous toxic-monitoring of wastewater influent is needed. This paper describes the potential toxic monitoring applications of the toxicity monitor using a nitrifying bacteria biosensor to sewerage systems. The results of sensitivity tests show that aspects of wastewater do not affect the sensor sensitivity and confirm that the sensor can be applied to wastewater monitoring as it is. The monitor with a prototype of filtration system installed in a wastewater treatment plant is able to operate continuously for one month at least after the modification of filtration system and the optimization of operation conditions. PMID- 11936644 TI - Real time control of a combined UASB-activated sludge wastewater treatment configuration. AB - This paper presents a combined wastewater treatment configuration composed by an Upflow Anaerobic Sludge Blanket (UASB) reactor followed by an Activated Sludge (AS) system. A control strategy has been developed for this innovative (especially for domestic sewage) treatment configuration and tested in a real pilot-scale system called STEPAA-Wastewater Treatment System by Anaerobic and Aerobic Processes. The proposed UASB-AS control strategy, including fault detection and recovery, and its successful implementation in real time is presented. This novel control strategy was developed to keep the final effluent suspended solids concentration in the range specified by the State environmental agency, in spite of incoming load disturbances. The control strategy is based on two cascaded PI (Proportional + Integral) controllers, which manipulates the recycling rate into the AS-reactor to control the effluent suspended solids concentration. A 2-dimensional nonlinear mapping (an empirical look-up table), which gives the sludge waste rate as a function of the influent flowrate and AS reactor biomass concentration, is used to keep the AS-reactor biomass concentration in a range that guarantees a good substrate removal without inconveniences to the AS-settler operation (and consequently to the solids removal). Experimental results are provided to demonstrate the system performance. PMID- 11936645 TI - A hybrid supervisory system to support WWTP operation: implementation and validation. AB - Integrated operation of Wastewater Treatment Plants is still far from being solved. A reasonable proposal should link advanced and robust control algorithms to some knowledge-based techniques, allocating the detailed engineering to numerical computations, while delegating the logical analysis and reasoning to supervisory intelligent systems. This paper describes the development and implementation of a knowledge-based Hybrid Supervisory System to support the operation of a real Wastewater Treatment Plant. The system integrates different reasoning modules, overcoming the limitations in the use of each single technique, while providing an agent based architecture with additional modularity and independence. It is structured into three separated levels: data gathering, diagnosis, and decision support. The different tasks of the system are performed in a seven-step cycle: data gathering and update, diagnosis, supervision, prediction, communication, actuation, and evaluation phase. In spite of certain reservations of the scientific community about the use of these techniques, the system is successfully performing real-time support to the operation of the Granollers facility since September 1999. Results of the first four-month validation period are shown and discussed. An example of the system behavior is also shown in the paper. The conclusions indicate the key steps which are necessary to transfer the system to another facility. PMID- 11936646 TI - Control of nitrate ricirculation flow in predenitrification systems. AB - The control of the nitrate recirculation flow in a predenitrification system is addressed. An elementary mass balance analysis on the utilisation efficiency of the influent biodegradable COD (bCOD) for nitrate removal indicates that the control problem can be broken down into two parts: maintaining the "anoxic" zone anoxic (i.e. nitrate is present throughout the anoxic zone) and maximising the usage of influent soluble bCOD for denitrification. Simulation studies using the Simulation Benchmark developed in the European COST program show that both objectives can be achieved by maintaining the nitrate concentration at the outlet of the anoxic zone at around 2 mgN/L. This setpoint appears to be robust towards variations in the influent characteristics and sludge kinetics. PMID- 11936647 TI - A framework for extreme-event control in wastewater treatment. AB - In this paper an approach to extreme event control in wastewater treatment plant operation by use of automatic supervisory control is discussed. The framework presented is based on the fact that different operational conditions manifest themselves as clusters in a multivariate measurement space. These clusters are identified and linked to specific and corresponding events by use of principal component analysis and fuzzy c-means clustering. A reduced system model is assigned to each type of extreme event and used to calculate appropriate local controller set points. In earlier work we have shown that this approach is applicable to wastewater treatment control using look-up tables to determine current set points. In this work we focus on the automatic determination of appropriate set points by use of steady state and dynamic predictions. The performance of a relatively simple steady-state supervisory controller is compared with that of a model predictive supervisory controller. Also, a look-up table approach is included in the comparison, as it provides a simple and robust alternative to the steady-state and model predictive controllers. The methodology is illustrated in a simulation study. PMID- 11936648 TI - A supervisory control system for optimising nitrogen removal and aeration energy consumption in wastewater treatment plants. AB - A fuzzy logic supervisory control system for optimising nitrogen removal and aeration energy consumption has been developed. This control system allows optimising and controlling the dissolved oxygen (DO) concentration in the aerobic reactors, the blowers discharge pressure and the effluent ammonia and nitrate concentrations. DO is controlled by adjusting control valve opening and blower discharge pressure is controlled by modifying rotational speed of the blowers. Optimum nitrification/denitrification is achieved by modifying the DO set point in the last aerobic reactor and the internal recirculation. This system has been tested by simulation in a Bardenpho process using the Activated Sludge Model No2. A significant improvement in stability on the activated sludge process is achieved. Moreover, significant energy saving has been achieved with this control strategy. PMID- 11936649 TI - A hedging point strategy--balancing effluent quality, economy and robustness in the control of wastewater treatment plants. AB - An operational space map is an efficient tool to compare a large number of operational strategies to find an optimal choice of setpoints based on a multicriterion. Typically, such a multicriterion includes a weighted sum of cost of operation and effluent quality. Due to the relative high cost of aeration such a definition of optimality result in a relatively high fraction of the effluent total nitrogen in the form of ammonium. Such a strategy may however introduce a risk into operation because a low degree of ammonium removal leads to a low amount of nitrifiers. This in turn leads to a reduced ability to reject event disturbances, such as large variations in the ammonium load, drop in temperature, the presence of toxic/inhibitory compounds in the influent etc. Hedging is a risk minimisation tool, with the aim to "reduce one's risk of loss on a bet or speculation by compensating transactions on the other side" (The Concise Oxford Dictionary (1995)). In wastewater treatment plant operation hedging can be applied by choosing a higher level of ammonium removal to increase the amount of nitrifiers. This is a sensible way to introduce disturbance rejection ability into the multi criterion. In practice, this is done by deciding upon an internal effluent ammonium criterion. In some countries such as Germany, a separate criterion already applies to the level of ammonium in the effluent. However, in most countries the effluent criterion applies to total nitrogen only. In these cases, an internal effluent ammonium criterion should be selected in order to secure proper disturbance rejection ability. PMID- 11936650 TI - Model structure identification for wastewater treatment simulation based on computational fluid dynamics. AB - The objective of this presented project is to use the results of an CFD simulation to automatically, systematically and reliably generate an appropriate model structure for simulation of the biological processes using CSTR activated sludge compartments. Models and dynamic simulation have become important tools for research but also increasingly for the design and optimisation of wastewater treatment plants. Besides the biological models several cases are reported about the application of computational fluid dynamics ICFD) to wastewater treatment plants. One aim of the presented method to derive model structures from CFD results is to exclude the influence of empirical structure selection to the result of dynamic simulations studies of WWTPs. The second application of the approach developed is the analysis of badly performing treatment plants where the suspicion arises that bad flow behaviour such as short cut flows is part of the problem. The method suggested requires as the first step the calculation of fluid dynamics of the biological treatment step at different loading situations by use of 3-dimensional CFD simulation. The result of this information is used to generate a suitable model structure for conventional dynamic simulation of the treatment plant by use of a number of CSTR modules with a pattern of exchange flows between the tanks automatically. The method is explained in detail and the application to the WWTP Wuppertal Buchenhofen is presented. PMID- 11936651 TI - Respirometry-based on-line model parameter estimation at a full-scale WWTP. AB - This paper describes part of a project to develop a systematic approach to knowledge extraction from on-line respirometric measurements in support of wastewater treatment plant control and operation. The paper deals with the following issues: (1) test of the implementation of an automatic set-up consisting of a continuous laboratory respirometer integrated in a mobile trailer with sampling and dosing equipment, and data-acquisition and communication system; (2) assessment of activated sludge/sewage characteristics from sludge respirograms by model parameter estimation; (3) comparison of the parameter estimates with regular plant data and information obtained from supplementary wastewater respirograms. The paper describes the equipment and some of its measuring results from a period of one week at a large-scale wastewater treatment plant. The measurements were evaluated in terms of the common activated sludge modelling practice. The automatic set-up allowed reliable measurements during at least one week. The data were used to calibrate two different version of the model, and independent parameter estimates were obtained. PMID- 11936652 TI - Optimal operation for timely adaptation of activated sludge plants to changes in the surfactant composition of wastewater. AB - The composition of a textile industry wastewater is highly variable, as the industrial process has to follow fashion and season trends. Surfactants represent one of the largest COD fractions in a typical textile wastewater. Therefore, it was the aim of this paper to model the acclimatisation behaviour of an activated sludge system when subjected to composition variations in the surfactant containing feed. The model was based on data obtained in SBR experiments in which a linear alkyl ethoxylate as sole carbon source in the feed was replaced by another with a longer ethoxylate chain. A previously developed model (Fractionated Degradation Model) was applied to each of the 21 SBR cycles carried out in this study. The resulting best-fit parameters were investigated and sub models were further developed, to create an acclimatisation model, able to predict the sludge acclimatisation level. Using the information given by this model, it was possible to propose an optimal operation scheme to pre-acclimatise the sludge before a surfactant replacement is made in the textile process. A cost analysis was carried out to compare different scenarios, with and without the application of this operation scheme. It was concluded that the proposed pre acclimatisation process may be cost effective as compared to other scenarios if a cheap surfactant-containing product was employed. PMID- 11936653 TI - Dynamic simulation of chemical industry wastewater treatment plants. AB - High variability, stringent effluent permits, and often extreme operating conditions define the practice of wastewater treatment in the chemical industry. This paper reviews the benefits and challenges of applying dynamic simulation to chemical-industry wastewater treatment plants by describing case studies at full scale wastewater treatment plants (WWTP). The applications range from process troubleshooting to optimization and control. The applications have been valuable and useful in developing a deeper understanding of the plants as integrated systems. However there still remains substantial work to implement the dynamic simulations for daily real-time use by plant engineers and operators. This opportunity to improve plant operations is still largely untapped and will remain so until dynamic state estimation and data reconciliation are incorporated into simulation packages for use in developing the on-line simulations. PMID- 11936654 TI - Improved design and optimization of aeration control for WWTPs by dynamic simulation. AB - This paper introduces a model library based on the simulation environment MATLAB/SIMULINK which allows the simulation of the aeration system of an wastewater treatment plant (WWTP). The model library presented covers all parts of an aeration system starting with the compressor set over the common air rail, flow resistances through pipes, controllable valves, aeration membranes and ending last not least with the controllers involved. The main objective of the development of the simulation system is the design, test and the support of the installation of more predictable and efficient control concepts for the aeration system in WWTPs. An example is given to demonstrate the application of the simulation system and to promote a control scheme with a flexible reference value for the pressure at the common air rail based on a most open valve (MOV) concept. PMID- 11936655 TI - Fuzzy predictive control for nitrogen removal in biological wastewater treatment. AB - Whenever the carbon/nitrogen ratio of a domestic wastewater is too low, full denitrification is difficult to obtain and an additional source of organic carbon has to be provided. Since loading conditions may vary appreciably over the diurnal cycle, depending on the weather and sewage conditions, dosing should be controlled by an adaptive regulator to keep into account the time-varying process dynamics. A fuzzy predictive controller is proposed in this paper and its performance is tested through numerical simulations. The new aspects brought forward are the use of an improved model for denitrification, the use of benchmark (i.e. thoroughly tested and standardised) input files and the conclusion about regulator performance in overall plant performance, in terms of carbon saving and discharge compliance. PMID- 11936656 TI - Model reduction through boundary relocation to facilitate real-time control optimisation in the integrated urban wastewater system. AB - Real time control is one of the possibilities to minimise the impact of the integrated urban wastewater system (sewer system and treatment plant) on the receiving water quality. Integrated control uses information about the river state to act in the sewer system or in treatment plant. In order to test and tune these integrated controllers, a simplified integrated model is needed. Even with these simplified models, the simulation times may be too long and further model reduction is needed. In this paper, dependency-structure based model reduction is proposed as a technique to further reduce model complexity. Three steps are proposed: relocation of the upstream system boundaries to just upstream of the first control point, relocation of the downstream boundaries to just downstream of the last measurement point, and third, a further model simplification based on an analysis of the sensitivity of the control actions on submodel elimination. The effect of applying the different reduction approaches on the control strategy and on the resulting river water quality is discussed on the basis of a case study of the catchment of Tielt. PMID- 11936657 TI - A new real-time drinking water network mixing model for the control and command centre of the city of Paris. AB - The Paris drinking water system has the particularity of being supplied by many different sources: surface water from the Seine and Marne rivers and underground water from sixty-three springs. This diversity of origins of the water and the existence of a gridded network leads the company in charge of the production and the quality of the water in Paris, Sagep, to partly mix the water in the network. Knowing the impact of the mixing on the physical parameters, and consequently on the quality of this mixed water, Sagep had searched for a tool to control the water origins in the network in order to limit extreme changes. A study led Sagep to develop a new kind of real-time water mixing model. PMID- 11936658 TI - Modelling and estimation of physical parameters in a sludge drying system. AB - In this paper is presented the study of a Sludge Drying System used to kill pathogenic organisms living in sludge. The system is modeled and the physical parameters thermal capacity, thermal resistance and thermal time constant are estimated using conventional estimation methods. PMID- 11936659 TI - Practical identification of the dissolved oxygen dynamic in activated sludge plants. AB - This paper presents the application to a full-scale waste water treatment plant (WWTP) of a practical methodology to identify the oxygen supply and consumption terms, which regulate the dynamic behaviour of the dissolved oxygen concentration (S(O)) in activated sludge reactors. This methodology is based on a periodic adjustment of the selected model for oxygen mass-transfer coefficient (K(L)a) and a continuous estimation of the oxygen uptake rate (r(O)), maintaining constant the adjusted K(L)a model, in order to uncouple the effects that the consumption and supply terms have on the S(O) concentration. The adjustment phase of the K(L)a model is based on typified excitations of the system through the aeration system of the plant, which allow the estimation of K(L)a for different values of the aeration intensity parameter, for instance, air flow rate in air diffusion systems, revolution speed in surface aerators, etc. Once the K(L)a model is adjusted, it is possible to evaluate the K(L)a value in process conditions at any time. The continuous estimation of r(O) is carried out starting from the oxygen mass balance in the activated sludge reactor. This practical methodology has been successfully verified in the Badiolegi WWTP in Azpeitia (Spain), which biological process consists of a double stage AB process, with two different aeration systems: fine pore air diffusers in the A-stage and surface aerators in the B stage. PMID- 11936660 TI - Genetic algorithms for the application of Activated Sludge Model No. 1. AB - The genetic algorithm (GA) has been integrated into the IWA ASM No. 1 to calibrate important stoichiometric and kinetic parameters. The evolutionary feature of GA was used to configure the multiple local optima as well as the global optimum. The objective function of optimization was designed to minimize the difference between estimated and measured effluent concentrations at the activated sludge system. Both steady state and dynamic data of the simulation benchmark were used for calibration using denitrification layout. Depending upon the confidence intervals and objective functions, the proposed method provided distributions of parameter space. Field data have been collected and applied to validate calibration capacity of GA. Dynamic calibration was suggested to capture periodic variations of inflow concentrations. Also, in order to verify this proposed method in real wastewater treatment plant, measured data sets for substrate concentrations were obtained from Haeundae wastewater treatment plant and used to estimate parameters in the dynamic system. The simulation results with calibrated parameters matched well with the observed concentrations of effluent COD. PMID- 11936661 TI - Simulation of a nitrification control concept considering influent ammonium load. AB - Buchenhofen wastewater treatment plant of Wupperverband (650,000 p.e.) is currently being expanded for targeted nitrogen elimination. In view of the limited space available for extension, an optimized control concept is to be used in order to minimize the number of additional tanks required. This concept was investigated by dynamic simulation based on Activated Sludge Model No. 1. The investigations included a pure feedback control configuration and a configuration combining feedback und feedforward control, considering the influent ammonium load, for aeration. The results show that combined feedforward/feedback control has significant advantages over pure feedback control. In particular, this configuration allows a reduction in the effluent NH4-N peaks, which is especially important because of the low NH4-N limit of 5 mg NH4-N/L in a grab sample. PMID- 11936662 TI - Online load measurement in combined sewer systems--possibilities of an integrated management of waste water transportation and treatment. AB - To obtain a further appreciable reduction of discharges in the area of sewage disposal, besides waste water purification at our treatment plants, discharge of wastewater through the sewer system has to be taken into account. Today, control strategies pursuing this aim are mainly based on hydraulic conditions like level or flow rate. They all neglect the wastewater organic load as an essential parameter. The main reasons are the expensive methods used to continuously measure traditional organic sum parameters like COD or TOC. A meaningful alternative to those parameters is the spectral absorption coefficient at lambda = 254 nm (SAC), defined in DIN 38402 by the German Institute for Standardisation. As a purely physical parameter, the SAC shows a good correlation to organic sum parameters like COD and TOC, especially if municipal wastewater is considered. By using an UV-process probe, it is possible to measure the SAC and infer the organic load of raw wastewater continuously without any sample pre-treatment. By the use of this instrument numerous possibilities arise, in order to control the sewers discharge load depend. PMID- 11936663 TI - Dynamic online sewer modelling in Helsingborg. AB - Within the last decade, the sewer system in Helsingborg, Sweden has been rehabilitated in many ways along with the reconstruction of the WWTP Oresundsverket in order to obtain a high degree of nitrogen and phosphorus removal. In that context a holistic view has been applied in order to optimise the corrective measures as seen from the effects in the receiving waters. A sewer catchment model has been used to evaluate several operation strategies and the effect of introducing RTC. Recently, a MOUSE ONLINE system was installed. In this phase the objective is to establish a stable communication with the SCADA system and to generate short-term flow forecasts. PMID- 11936664 TI - Evaluation of uncertainties in urban hydrology: application to volumes and pollutant loads in a storage and settling tank. AB - It is necessary to assess adequately measurement uncertainties of experimental data in urban hydrology in order to draw pertinent and valuable conclusions from measurement results. This paper first gives a brief presentation of the concepts relative to standard uncertainties and to the law of propagation of uncertainties. Then it presents an example of calculation of the uncertainties in some quantities describing the behaviour of the Venissieux (France) storage and settling tank during one rainfall event. These quantities are the flow rates and the volumes entering into the tank, the inflow and outflow mean concentrations and masses of TSS (Total Suspended Solids), and the TSS removal rate. Variographs are used to account for the autocorrelation of time series data. The results lead to the following relative uncertainties 8% for the total volume, 30% for the inflow concentration, 38% for the outflow concentration, 31% for the inflow mass, 39% for the outflow mass and 138% for the removal rate. PMID- 11936665 TI - Feasibility of automatic chemicals dosage control--a full-scale evaluation. AB - This contribution discusses the feasibility of automatic control for chemicals dosing in activated sludge systems. The evaluation is made on the basis of a full scale implementation at the Lommel WWTP (Belgium) of an on-line controlled dosage system for iron chloride and external carbon source. The control laws are very simple and allow intuitive adaptation by the plant operators. The control system results in a significant reduction of the chemicals dosage, better effluent results and a lower sludge production. The implementation is furthermore cost efficient. PMID- 11936666 TI - Control of the aeration volume in an activated sludge process for nitrogen removal. AB - Biological nitrogen removal in an activated sludge process is obtained by two biological processes; nitrification and denitrification. Nitrifying bacteria need dissolved oxygen and a sufficiently large aeration volume for converting ammonium to nitrate in the wastewater. The objective of this paper is to develop an automatic control strategy for adjusting the aerated volume so that the effluent ammonium level can be kept close to a desired value despite major changes in the influent load. The strategy is based on applying exact linearization of the IAWO Activated Sludge Process Model No 1. Simulation results show that the suggested controller effectively attenuates process disturbances. PMID- 11936668 TI - Remote control system for the operation of 65 WWTPs. AB - The present report discusses the monitoring system for approximately 340 facilities for water management of the Emschergenossenschaft and Lippeverband. It is based on the operation control system of the wastewater treatment plants (currently 61) and the pumping stations of the Lippeverband, which has been created since 1985. For the near future, the integration of the plants of the Emschergenossenschaft including storm-water tanks is scheduled. The current connection ratios of the WWTPs which are to be monitored vary between 500 and 1,940,000 Population Equivalents. The data recorded at each plant are transmitted to the allocated catchment area centre and then to the district centres. The encompassing monitoring centre serves the integrated control of all plants. The comprehensive remote control is managed through the presentation of relevant plant-type-specific system parameters. The investment costs of approximately 33,000 Euros/plant and annual costs of 9,400 Euros/plant stand against a considerable increase of the operation safety and the reduction of manpower. PMID- 11936667 TI - Dissolved oxygen controller based on on-line measurements of ammonium combining feed-forward and feedback. AB - As the largest single energy-consuming component in most biological wastewater treatment systems, control of aeration is of great interest seen from an energy savings point of view. This paper suggests a simple way of using on-line ammonium measurements to control aeration in a pre-denitrification plant by controlling the dissolved oxygen setpoint. The controller works primarily by feed-forward based on an ammonium sensor located at the head of the aerobic process part. Using online in-situ sensor measurements directly from the process have the important advantage over effluent measurements that there is no or very short time delay for information. The controller has been implemented in a full-scale wastewater treatment plant for a period of 35 days. During the experiment two identical activated sludge lines were used. The controller was implemented in one line, while the other line worked as a reference for comparison. The preliminary results indicate that the described control strategy leads to energy savings for the aeration in the region of 5-15%, while maintaining approximately the same effluent quality as in the reference line. Even higher energy savings can probably be achieved by optimising the controller. An automatic procedure for updating the controller parameters based on dynamic effluent ammonium measurement has been tested. PMID- 11936669 TI - On-line instrument confirmation: how can we check that our instruments are working? AB - As on-line instrumentation becomes increasingly important for operating modern wastewater treatment plants it is necessary to have diagnostic methods that can indicate sensor deviation at an early stage. Examples of on-line diagnosis methods used at the Rya WWTP (Goteborg, Sweden) are presented. These include on line mass balance calculations and soft sensors, which make indirect parameter estimates. By comparing the estimates with the corresponding on-line sensors it is possible to make control decisions and to chose alternative strategies. These comparative methods are often useful for process monitoring and diagnosis as well. Some other soft sensor applications using exponential filters are also presented. PMID- 11936670 TI - International online support to process optimisation and operation decisions. AB - The information level at all technical facilities has developed from almost nothing 30-40 years ago to advanced IT--Information Technology--systems based on both chemical and mechanical on-line sensors for process and equipment. Still the basic part of information is to get the right data at the right time for the decision to be made. Today a large amount of operational data is available at almost any European wastewater treatment plant, from laboratory and SCADA. The difficult part is to determine which data to keep, which to use in calculations and how and where to make data available. With the STARcontrol system it is possible to separate only process relevant data to use for on-line control and reporting at engineering level, to optimise operation. Furthermore, the use of IT makes it possible to communicate internationally, with full access to the whole amount of data on the single plant. In this way, expert supervision can be both very local in local language e.g. Polish and at the same time very professional with Danish experts advising on Danish processes in Poland or Sweden where some of the 12 STARcontrol systems are running. PMID- 11936671 TI - Status and future trends of ICA in wastewater treatment--a European perspective. AB - The status of instrumentation, control and automation (ICA) within the European wastewater community is reviewed and some major incentives and bottlenecks are defined. Future trends of ICA are also discussed. The information is based on a COST 624 workshop and a non-exhaustive survey with regard to ICA carried out in 13 European countries during March 2001. The level of instrumentation (type of sensors, usage frequency, etc.) and how these instruments are used for on-line control purposes are presented for each individual country (Austria, Belgium, Czech Republic, Denmark, Finland, France, Germany, Netherlands, Romania, Slovenia, Spain, Sweden and Switzerland). The most common types of applied real time control in wastewater treatment plants are given. One conclusion of the paper is that sensors no longer represent the main bottleneck for on-line control, rather the lack of plant flexibility is more troublesome. Moreover, the current transitional phase of the wastewater industry in Europe represents a unique opportunity to apply ICA on a large scale. The driving forces are simply too strong to ignore. PMID- 11936672 TI - Evaluation of a four year experience with a fully instrumented anaerobic digestion process. AB - For several years, a 1 m3 fixed bed anaerobic digestion process has been operated for the treatment of distillery vinasses. This reactor has been fully instrumented with the following variables available on-line: pH, temperature, liquid and gas flow rates, gas composition (i.e., CH4, CO2 and H2), concentration of bicarbonate, chemical oxygen demand, total organic carbon, volatile fatty acids and partial and total alkalinity, these last four variables being measured twice by different techniques (i.e., using a TOC analyzer, a titrimetric sensor and an infrared spectrometer). The purpose of this paper is to compare the respective benefits of advanced instrumentation for the monitoring of wastewater treatment processes in general, and for anaerobic digestion in particular. It will also provide some statistical analysis of the time required to operate a fully instrumented wastewater treatment process. It is indeed well admitted in the literature that instrumentation is usually the main limitation step for using closed-loop control. However, it is our opinion that, in the near future, this situation will change. This point is discussed based on our four years practical experience. PMID- 11936673 TI - Implementation of a wastewater treatment plant operation support tool based on on line simulation. AB - The operators of modern wastewater treatment plants (WWTPs) are faced with increasing demands. Beyond the requirement to consistently meet discharge limits for pollutant loads, the cost efficiency of plant operation is becoming more and more important. This results in new challenges for automatic control and human control action. On-line simulation opens up interesting perspectives to provide comprehensive process information, serving as a base for optimised operation of WWTPs. This paper describes the development and application of a computer analysis and support tool for the large-scale municipal WWTP of the city of Magdeburg, Germany. It will show that by linking a simulation server to the Distributed Control System (DCS) relevant additional features for plant control arise. A good cost-benefit ratio of the system is achieved by using web techniques for implementing the software. PMID- 11936674 TI - Diagnosing and optimizing water treatment processes by using particle counter: a case study in Korea. AB - The goal of the flocculation process is to change the particle size distribution to best suit the subsequent processes. Although several methods exist to evaluate the flocculation process, no single universally accepted method has yet to be developed. The purpose of this paper is to present experiences whereby particle counting was used in the diagnosis and optimization of the flocculation process. A commercially available on-line continuous particle counter has been used in evaluating the design and the operation of this process at two conventional Water Treatment Plants. The evaluation is based on particle dynamics, i.e., the change of the number of small and large particles. Some design deficiencies in the distribution channel and flocculation process have been identified from this method, and thus some operational parameters are suggested for optimum performance. Because the optimum condition may be site-specific, the method presented in this paper will be beneficial in the evaluation of the flocculation process at other water treatment plants. PMID- 11936675 TI - Cascade control strategy for external carbon dosage in predenitrifying process. AB - We propose a cascade control strategy composed of two Proportional-Integral (PI) controllers to regulate the nitrate concentration in the predenitrifying process by manipulating the external carbon dosage. It controls the nitrate concentrations in the effluent as well as in the final anoxic reactor simultaneously to strictly satisfy the quality of the effluent as well as to remove the effects of disturbances more quickly. The design of two PI controllers in the cascade control loop can be completed with the Ziegler-Nichols (Z-N) tuning rule together with a simple relay feedback identification method. Results from the Benchmark simulation confirm that both good set point tracking and satisfactory disturbance rejection can be guaranteed due to the structural advantages of the proposed cascade control strategy. Also, compared with a previous work, the fluctuation of the nitrate concentration in the effluent has been decreased significantly. PMID- 11936676 TI - On-line titration of non-ionic surfactants in wastewater treatment plants using a specific electrode. AB - Textile finishing industry wastewater often contains high concentrations of surfactants. Non-ionic surfactants like alcohol ethoxylates are among the most used surfactants and are discharged from batch-processes in widely varying concentrations. The anaerobic biomass and especially methanogenic microorganisms have been shown to be inhibited by surfactants. To protect these microorganisms from irreversible inhibition a measurement of the surfactant concentration is useful for the process control, but most analytical methods are too complicated and expensive for an on-line process control. It was shown that the titration of non-ionic surfactants is possible in the presence of biomass and in wastewater from textile wet processes using the Metrohm NIO-electrode. Titration and sampling were successfully performed with standard equipment. The software used for the process control of a lab-scale anaerobic treatment plant was also used to evaluate the obtained titration curves. This allowed performing the titrations without using a more expensive titrator. PMID- 11936677 TI - On-line determination of nitrite in wastewater treatment by use of a biosensor. AB - A newly developed biosensor for nitrite having a 90% response time of about 1 min was used to monitor nitrite concentration in activated sludge exposed to oxic/anoxic cycles. The NO2- biosensor contains bacteria that reduce NO2-, but not NO3-, to N2O that is subsequently monitored by a built-in electrochemical sensor. Nitrite plus nitrate (NOx-) was simultaneously monitored by a NOx- biosensor. The maximum operational lifetime of the NO2- biosensor was 6 weeks, but much longer lifetimes can be expected as malfunctioning by the 3 sensors used for longer periods was due to either mechanical damage or ineffective internal sterilization during the construction. Insufficiently sterilized sensors became sensitive also to NO3- after some time due to development of NO3(-)-reducing bacterial populations within the sensor. The fraction of NO2- as compared to NO3- in the activated sludge was very dependent on prehistory, actual loading, and aeration. During balanced operation with NH4+ being exhausted during the later parts of the aerobic cycle, NO2- increased in concentration up to about 50 microM during the early part of the aeration cycle until NH4+ became limiting. At that time the NO2- concentration decreased to low levels. Under some operating conditions a peak of NO2- also appeared in the beginning of the anoxic period. NO2- and NO3- were depleted simultaneously during the anoxic period. PMID- 11936678 TI - Investigations of the dynamic behaviour of the composition of combined sewage using on-line analyzers. AB - Continuously analyzing sensors today permit the recording of the dynamic behaviour pattern of pollutants in combined sewage through physical parameters. This report presents the results of such measurements. They were done in a storm water tank which is equipped with a combined sewage overflow and located at the end of a 360 ha catchment area. The results show that by using a combination of sensors for dissolved solids (using UV absorption) and of sensors for particulate solids (using scattered light measurement), reliable information on the composition of the combined sewage can be obtained. A statistical relationship between these two parameters and the chemical oxygen demand was established. The analyzing methods presented permit the real-time control of sewer systems on the basis of the pollution carried in the combined sewage. Through the information on the actual concentration of primary pollutants provided by on-line analysis, heavily polluted streams can be retained selectively in storm water tanks or directed to the treatment plant. However, due to the adverse conditions in a sewer system operational disturbances and stringent maintenance requirements are still an impediment to long-term operation. PMID- 11936679 TI - Evaluation of different shape parameters to distinguish between flocs and filaments in activated sludge images. AB - The ratio of flocs to filaments in activated sludge waste water treatment plants is of extreme importance for the overall performance of the plant. In order to control this ratio the individual concentrations of flocs and filaments need to be measurable. However, no sensors which can measure these concentrations are currently available. It is proposed that by means of image analysis techniques the ratio of flocs to filaments can be determined. Combination of this ratio with the total biomass concentration results in the individual floc and filament concentration. This contribution focuses on the last step of the image analysis procedure, i.e., the classification of objects as either floc or filament. Five different shape parameters, i.e., aspect ratio, roundness, form factor, fractal dimension and reduced radius of gyration, are evaluated and compared. The results indicate that the form factor is the least suitable and the reduced radius of gyration the most suitable shape parameter to accurately classify flocs and filaments in activated sludge images. PMID- 11936680 TI - Model-based advanced process control of coagulation. AB - The drinking water treatment industry has seen a recent increase in the use of artificial neural networks (ANNs) for process modelling and offline process control tools and applications. While conceptual frameworks for integrating the ANN technology into the real-time control of complex treatment processes have been proposed, actual working systems have yet to be developed. This paper presents development and application of an ANN model-based advanced process control system for the coagulation process at a pilot-scale water treatment facility in Edmonton, Alberta, Canada. The system was successfully used to maintain a user-defined set point for effluent quality, by automatically varying operating conditions in response to changes in influent water quality. This new technology has the potential to realize significant operational cost saving for utilities when applied in full-scale applications. PMID- 11936681 TI - In-situ measurement of ammonium and nitrate in the activated sludge process. AB - A new in-situ probe is presented for the continuous measurement of ammonium and nitrate in wastewater. It requires no sample preparation and is installed directly in the process liquid. This new low-cost probe significantly reduces investment and operating costs and requires minimum maintenance. The paper describes the sensor principle and test results from three different probe locations: the primary clarifier effluent, the activated sludge tank and the nitrifying biofilter influent. Reference measurements were carried out by means of conventional analyzers with ultrafiltration, an in-situ UV spectrometer for the nitrate and laboratory analysis of spot and 2h-composite samples. The aim of the study was to investigate the operational reliability and accuracy of the new probe and the expenditure required for its maintenance and calibration. The tests showed that the new probe performed very well overall and required minimum maintenance. Some problems were observed during the biofilter plant test. They are assumed to be related to substantial changes in the wastewater composition. PMID- 11936682 TI - Determination of carbamazepine and carbamazepine 10,11-epoxide in human plasma by tandem liquid chromatography-mass spectrometry with electrospray ionisation. AB - A sensitive method for the determination of carbamazepine and carbamazepine 10,11 epoxide in plasma is described, using high-performance liquid chromatographic separation with tandem mass spectrometry. Samples were purified using liquid liquid extraction and separated on a Phenomenex Luna C18 5 microm. 150 x 2 mm column with a mobile phase consisting of acetonitrile, methanol and formic acid (0.1%) (10:70:20, v/v). Detection was performed by a Micromass Quattro Ultima mass spectrometer in the MRM mode (LC-MS-MS) using electro spray ionisation (ESI+), monitoring the transition of the protonated molecular ion for carbamazepine at m/z 237.05 and carbamazepine 10,11-epoxide at m/z 253.09 to the predominant ions of m/z 194.09 and 180.04, respectively. The mean recovery was 95% for carbamazepine and 101% for carbamazepine 10,11-epoxide, with a lower limit of quantification of 0.722 ng/ml for carbamazepine and 5.15 ng/ml for carbamazepine 10,11-epoxide, when using 0.5 ml plasma. This high-throughput method was used to quantify 230 samples per day, and is sufficiently sensitive to be employed in pharmacokinetic studies. PMID- 11936683 TI - Determination of triamterene and its main metabolite hydroxytriamterene sulfate in human urine by capillary electrophoresis using ultraviolet absorbance and laser-induced fluorescence detection. AB - Two capillary electrophoresis methods have been developed for the direct determination of triamterene and its main metabolite hydroxytriamterene sulfate in human urine. Analytes were detected using conventional UV detection as well as laser-induced fluorescence (LIF) detection with an HeCd-laser operating at a wavelength of 325 nm. The results of both detection techniques were compared. Indeed, the limit of quantification was eightfold lower using LIF detection (50 ng/ml) in comparison to UV detection (400 ng/ml). As no interference due to endogenous urine compounds was observed, direct urine analysis was feasible. Analysis was very simple and fast-one run could be performed within less than 10 min (CE-UV method) and 2.5 min (CE-LIF method), respectively. Both assays were fully validated and applied to urine samples from a human volunteer. The results of the application of the CE-LIF method to human urine samples are presented in this publication. PMID- 11936684 TI - Determination by high-performance liquid chromatography of phenylbutazone in samples of plasma from fighting bulls. AB - The purpose of this study was to investigate the possible presence of phenylbutazone in plasma samples from fighting bulls killed in 2nd and 3rd category bullrings in the province of Salamanca (Spain) in 1998, 1999 and 2000. For quantitative and qualitative determination, a high-performance liquid chromatograph was used, equipped with a photodiode-array detector and setting wavelengths at 240, 254 and 284 nm. The mobile phase optimized for the simultaneous detection of dexamethasone, betamethasone, flunixin and phenylbutazone, was 0.01 M acetic acid pH 3 in methanol (35:65 v/v) at a flow rate of 1 ml/min. Plasma samples were deproteinized with 400 microl of acetonitrile and 20 microl of the supernatant were injected directly into the chromatographic system equipped with a Lichrospher 60 RP select B column and guard column. For the quantitative analysis, standard calibration curves were made in a concentration range between 0.25 and 30 microg/ml, using betamethasone as internal standard. The retention time of phenylbutazone was 8.7 +/- 0.2 min and recovery was 83%. The detection and quantification limits were 0.016 and 0.029, respectively for A=240 nm. The study results show that 17 of the 74 samples analyzed in 1998, 18 of those from 1999 and 10 of those from 2000 were positive for phenylbutazone. PMID- 11936686 TI - Strategies for the purification and on-column cleavage of glutathione-S transferase fusion target proteins. AB - In this report, we describe a flexible, efficient and rapid protein purification strategy for the isolation and cleavage of glutathione-S-transferase (GST) fusion proteins. The purification and on-column cleavage strategy was developed to work for the purification of difficult proteins and for target proteins where efficient fusion-tag cleavage is essential for downstream processes, such as structural and functional studies. To test and demonstrate the flexibility of this method, seven diverse unrelated target proteins were assayed. A purification technique is described that can be applied to a wide range of both soluble and membrane inserted recombinant target proteins of differing function, structure and chemical nature. This strategy is performed in a single chromatographic step applying an on-column cleavage method, yielding "native" proteins in the 200 microg to 40 mg/l scale of 95-98% purity. PMID- 11936685 TI - Rapid determination of tafenoquine in small volume human plasma samples by high performance liquid chromatography-tandem mass spectrometry. AB - A method was developed for the determination of tafenoquine (I) in human plasma using high-performance liquid chromatography-tandem mass spectrometry. Prior to analysis, the protein in plasma samples was precipitated with methanol containing [2H3(15N)]tafenoquine (II) to act as an internal standard. The supernatant was injected onto a Genesis-C18 column without any further clean-up. The mass spectrometer was operated in the positive ion mode, employing a heat assisted nebulisation, electrospray interface. Ions were detected in multiple reaction monitoring mode. The assay required 50 microl of plasma and was precise and accurate within the range 2 to 500 ng/ml. The average within-run and between-run relative standard deviations were < 7% at 2 ng/ml and greater concentrations. The average accuracy of validation standards was generally within +/- 4% of the nominal concentration. There was no evidence of instability of I in human plasma following three complete freeze-thaw cycles and samples can safely be stored for at least 8 months at approximately -70 degrees C. The method was very robust and has been successfully applied to the analysis of clinical samples from patients and healthy volunteers dosed with I. PMID- 11936687 TI - Novel and sensitive high-performance liquid chromatographic method based on electrochemical coulometric array detection for simultaneous determination of catecholamines, kynurenine and indole derivatives of tryptophan. AB - A novel and simple method has been developed for the simultaneous quantification of tryptophan, kynurenine and indole derivatives as well as four catecholamines, including dopamine, noradrenaline, homovanillic acid and 3,4 dihydroxyphenylacetic acid. The method utilises isocratic reversed-phase high performance liquid chromatography with electrochemical coulometric array detection. The influence of various parameters on chromatographic performance, such as the composition and the pH of the mobile phase and the detection potentials, was investigated. Separation of 13 compounds was achieved by a mobile phase consisting of 10% methanol in 50 mM sodium phosphate-acetate buffer, pH 4.10, containing 0.42 mM octanesulphonic acid. The calibration curve was linear over the range 12 pg to 300 ng on-column. The detection limits (SIN 3) depended on the working potential and were found to be between 10 and 100 pg injected. The method was reproducible with intra-day RSDs of 0.3 to 1.5% and inter-day RSDs of 0.5 to 4%. PMID- 11936688 TI - Human blood and environmental media screening method for pesticides and polychlorinated biphenyl compounds using liquid extraction and gas chromatography mass spectrometry analysis. AB - Screening assessment methods have been developed for semi- and non-volatile persistent organic pollutants (POPs) for human blood and solid environmental media. The specific methodology is developed for measuring the presence of "native" compounds, specifically, a variety of organochlorine pesticides (OCPs), organophosphate pesticides (OPPs), and for polychlorinated biphenyls (PCBs). The method is demonstrated on anonymous Red Cross blood samples as well as two potential environmental sources, tracked in soil and dog hair. This work is based on previously developed methods for semi-volatile hydrocarbon exposure from fuels usage and similarly employs liquid solvent extraction, evaporative volume reduction. and subsequent specialized gas chromatography-mass spectrometry analysis (GC-MS). Standard curves, estimates of recovery efficiency, and specific GC-MS SIM quantification methods were developed for common pesticides including diazinon. aldrin, chlorpyrifos, malathion, dieldrin, DDT, permethrin, cyhalothrin, and cypermethrin, and for seven selected PCBs. Trace levels of certain PCBs and pesticides such as permethrin, dieldrin, malathion, lindane, diazinon, and chlorpyrifos were tentatively identified in anonymous blood samples as well as in two potential environmental sources. tracked in soil and dog hair. The method provides a simple screening procedure for various media and a variety of common organic pollutants without extensive sample preparation. It is meant to complement and augment data from more specific or complex methodology, to provide initial broad spectrum guidance for designing targeted experiments, and to provide confirmatory evidence for the usual metabolic biomarker measurements made to assess human exposure. PMID- 11936689 TI - Measurement of endogenous uracil and dihydrouracil in plasma and urine of normal subjects by liquid chromatography-tandem mass spectrometry. AB - A sensitive and specific HPLC-MS-MS method was developed for the determination of endogenous uracil (Ura) and its metabolite dihydrouracil (UH2) in human plasma and urine samples. Plasma samples were extracted with ethyl acetate-isopropanol (85:15, v/v) following added ammonium sulfate, and then separated on a Discovery Amide C16 column with 3% methanol solution as the mobile phase; urine samples were just centrifuged at 2500 g for detection. Quantitation was carried out by LC MS-MS in the multiple reaction monitoring (MRM) mode. The limits of quantitation of the method for Ura and UH2 were 0.5 and 5 ng ml(-1) (for plasma), and 50 and 100 ng ml(-1) (for urine), respectively. This method can be useful to evaluate the activity of dihydropyrimidine dehydrogenase (DPD), a rate-limiting enzyme of the chemotherapy drug fluoropyrimidine, which will be helpful in investigating subject variation of DPD and adjusting clinical dosage in pyrimidine chemotherapy. PMID- 11936690 TI - Sensitive and simple method for the determination of nicotine and cotinine in human urine, plasma and saliva by gas chromatography-mass spectrometry. AB - A method is proposed for the determination of nicotine and cotinine in human urine, plasma and saliva. Nicotine and cotinine were extracted from alkalinized sample with ethyl ether and concentrated to minimum volume with nitrogen stream. The volatility of nicotine was prevented by the addition of acetic acid to the organic solvent during evaporation. Peak shapes and quantitation of nicotine and cotinine are excellent, with linear calibration curves over a wide range of 1 10,000 ng/ml. The detection limits of nicotine and cotinine are 0.2 ng/ml in urine and 1.0 ng/ml in plasma and saliva. The intra-day precision of nicotine and cotinine in all samples was <5% relative standard deviation (RSD). Urine, plasma and saliva samples of 303 non-smoking and 41 smoking volunteers from a girl's high school in Korea were quantified by the described procedure. As a result, the concentrations of nicotine and cotinine in plasma ranged from 6 to 498 ng/ml and 4 to 96 ng/ml. Otherwise, those of nicotine and cotinine in saliva ranged from 0 to 207 ng/ml and 0 to 42 ng/ml, and those of nicotine and cotinine in urine ranged from 0 to 1,590 ng/ml and 0 to 2,986 ng/ml, respectively. We found that the concentration of cotinine in plasma was successfully predicted from the salivary cotinine concentration by the equation y=2.31x+4.76 (x=the concentration of cotinine in saliva, y=the concentration of cotinine in plasma). The results show that through the accurate determination of cotinine in saliva, the risk of ETS-exposed human can be predicted. PMID- 11936691 TI - Determination of the cell lytic properties of amphiphilic inhibitors of the cytosolic phospholipase A2 against human platelets by measuring the liberation of serotonin with high-performance liquid chromatography and fluorescence detection. AB - A procedure for the determination of the influence of detergents on the cell integrity of human platelets by measuring the release of serotonin with high performance liquid chromatography and fluorescence detection is presented. After exposure of the platelets to the test compounds the cells and cell fragments, respectively, are centrifuged off. In the supernatants the liberated serotonin is determined directly without a further sample clean up. The amphiphilic inhibitors of cytosolic phospholipase A2 (cPLA2), arachidonyltrifluoromethyl ketone (AACOCF3), palmityltrifluoromethyl ketone (PACOCF3) and methyl arachidonylfluorophosphonate (MAFP), and the polyoxyethylene detergent Brij 58 were investigated for their cell lytic properties with this method. All compounds lysed the platelets liberating serotonin at a concentration of 33 microM. AACOCF, and Brij 58 even caused cell lysis at lower concentrations. PMID- 11936693 TI - Metabolism of cymoxanil and analogs in strains of the fungus Botrytis cinerea using high-performance liquid chromatography and ion-pair high-performance thin layer chromatography. AB - The metabolism of cyano-oxime fungicide 1-(2-cyano-2-methoxyiminoacetyl)-3 ethylurea (cymoxanil) and analogs was studied on several strains of the fungus Botrytis cinerea owing to their difference in sensitivity towards cymoxanil. Chromatographic analysis of the unextracted culture medium was simpler and more accurate, particularly for ionizable metabolites because it avoids problems associated with extraction. Reversed-phase high-performance liquid chromatography was applied to compare the decrease of cymoxanil and analogs caused by different strains of B. cinerea, by periodic injections of incubated culture medium aliquots, directly on a C4 wide-pore column. Furthermore, a thin-layer chromatographic monitoring on C18 bonded silica gel with ion-pairing allowed the monitoring of the ionizable metabolites for substrates that were demonstrated to decompose most rapidly. These complementary analyses showed that the sensitivity of the highly sensitive strain towards cymoxanil was related to the disappearance of cyano-oximes studied from culture medium, namely to the ability of the strain B. cinerea to metabolize them. PMID- 11936692 TI - Determination of sodium 3,4-diaminonaphthalene-1-sulfonate, a Congo Red derivative, in plasma and brain of hamsters by high-performance liquid chromatography after solid-phase extraction and ultraviolet absorbance. AB - In the search for compounds with similar or greater activity than Congo Red (CR) in protecting normal prion protein from being converted into the pathological form, we have synthesized various compounds which derive from CR. One of these is the sodium 3,4-diaminonaphthalene-1-sulfonate (RCA) which has an activity similar to CR in preliminary experiments. This study describes a method to determine RCA in plasma and in brain tissue by high-performance liquid chromatography (HPLC), using a solid-phase extraction and UV detection. RCA is an amphoteric molecule difficult to separate from biological matrices. Extraction was achieved by solid phase extraction (ENV+ columns) together with the use of a counter ion. The resulting solid-phase extraction is efficient and rapid. RCA was separated on a Symmetry C18 250 x 4.6 mm I.D. 5 lm column at 1 ml/min using a 50 mM NaSO4 in 5 mM tetra-n-butylammoniumiodide (TEBA) in water-methanol (82:18, v/v) mobile phase. Retention times of RCA and I.S. were 21 and 24 min. The UV detector was set at 210 nm. The limit of quantitation was 0.5 microg/ml. The method has intra assay and inter-assay accuracies higher than 95%, coefficients of variation ranging between 2.8 and 8.6%, and recovery rates between 74.3 and 80.1% in plasma and in brain tissue. A linear response to quantities of RCA from 0.5 to 100 microg/ml or 10 microg/g in plasma or brain was obtained. The present method allows the study of the pharmacokinetics of RCA in plasma after i.p. administration, and the distribution of the compound into the brain at the peak time. PMID- 11936694 TI - Reversible fluorescence labeling of amino groups of protein using dansylaminomethylmaleic anhydride. AB - The reversible fluorescence labeling of insulin, catalase and lysozyme has been demonstrated. As a derivatizing reagent, dansylaminomethylmaleic acid (DAM) has been used after investigating the precolumn and precapillary derivatization conditions. This reagent (DAM) reacts with the amino groups of proteins via its anhydride in the presence of a suitable dehydrating reagent, which then could be liberated under mild acidic conditions and the native proteins are regenerated. After the derivatization of insulin, catalase and lysozyme with DAM, no peaks of these native proteins were observed while several peaks of the derivatized proteins due to the multiple labeling were observed. However, after the regeneration, increasing amounts of the native proteins were observed as the regeneration period increased. For the lysozyme, the bacteriolytic activity of the enzyme decreased after the derivatization, and only 0.9% of the activity remained. The activity increases by the regeneration, and 95.6% of the bacteriolytic activity of the native enzyme was observed after a 48-h regeneration at pH 2.5 and 40 degrees C. PMID- 11936695 TI - Simultaneous determination of hydrocodone and hydromorphone in human plasma by liquid chromatography with tandem mass spectrometric detection. AB - A rapid, sensitive and specific liquid chromatography-tandem mass spectrometry (LC-MS-MS) method has been developed and validated for the simultaneous analysis of hydrocodone (HYC) and its metabolite hydromorphone (HYM) in human plasma. A robotic liquid handler and a 96-channel liquid handling workstation were used to aliquot samples, to add internal standard (I.S.), and to extract analytes of interest. A 96-well mixed-mode solid-phase cartridge plate was used to extract the analytes and I.S. The chromatographic separation was on a silica column (50 x 3 mm, 5-microm) with a mobile phase consisting of acetonitrile, water and trifluoroacetic acid (TFA) (92:8:0.01, v/v). The run time for each injection was 2.5 min with the retention times of approximately 2.1 and 2.2 min for HYC and HYM, respectively. The tandem mass spectrometric detection was by monitoring singly charged precursor-->product ion transition 300-->199 (m/z) for HYC, and 28 ->185 (m/z) for HYM. The validated calibration curve range was 0.100-100 ng/ml, based on a plasma volume of 0.3 ml. The correlation coefficients were greater than or equal to 0.9996 for both HYC and HYM. The low limit of quantitation (LLOQ) was 0.100 ng/ml for both HYC and HYM with signal-to-noise ratio (S/N) of 50 and 10. respectively. The deuterated analytes, used as internal standards, were monitored at mass transitions 303-->199 (m/z) for HYC-d3 and 289-->185 (m/z) for HYM-d3. The inter-day (n= 17) precision of the quality control (QC) samples were < or = 3.5% RSD (relative standard deviation) for HYC and < or = 4.7% RSD for HYM, respectively. The inter-day accuracy of the QC samples were < or = 2.1% RE (relative error) for HYC and < or = 1.8% RE for HYM. The intra-day (n=6) precision and accuracy of the QC samples were < or = 2.6% RSD and < or = 3.0% RE for HYC, and < or = 4.7% RSD and < or = 2.4% RE for HYM. There was no significant deviation from the nominal values after a 5-fold dilution of high concentration QC samples by blank matrix. The QC samples were stable when kept at room temperature for 24-h or experienced three freeze-thaw cycles. The extraction recoveries were 86% for HYC and 78% for HYM. No detectable carryover was observed when a blank sample was injected immediately after a 2500 ng/ml sample that was 25-fold more concentrated than the upper limit of quantitation (ULOQ). PMID- 11936696 TI - Headspace solid-phase microextraction in combination with gas chromatography and tandem mass spectrometry for the determination of organochlorine and organophosphorus pesticides in whole numan blood. AB - A method for the determination of several organochlorine and organophosphorus pesticides in human whole blood samples was developed. The combination of solid phase microextraction in headspace mode with gas chromatography with tandem mass spectrometry allowed the determination of 11 selected pesticides at ppb levels, minimizing the sample treatment. Quantitation was carried out by means of calibration curves prepared in blood using labelled surrogate/internal standards. The method showed good linearity between 1 and 50 ng ml(-1) (0.5-25 ng ml(-1) for HCB) using second-order calibration curves. Precision was found to be better than 20% at the three concentration levels assayed in the range of ng ml(-1). The detection limits obtained were in the range 0.02-0.7 ng ml(-1), except for p,p' DDT (3 ng ml(-1)). The developed procedure was applied to blood and serum samples obtained from agricultural workers. HCB. beta-HCH and p,p'-DDE were most frequently detected in the samples analyzed. PMID- 11936697 TI - Quantification of perphenazine in eurasian otter (Lutra lutra lutra) urine samples by gas chromatography-mass spectrometry. AB - Perphenazine enanthate has been used in wild animals as a tranquilizer during the period of adaptation to new environments to reduce stress, mortalities and injuries. A gas chromatographic procedure for the quantitative measurement of perphenazine in otter urine has been developed and validated. The method involved an enzymatic hydrolysis with beta-glucuronidase-arylsulfatase from Helix pomatia, followed by a solid-phase extraction with Bond Elut Certify cartridges. The resulting organic phase was evaporated, and the dry extract was derivatised with MSTFA to form the O-TMS derivative. The derivatised extracts were analysed by gas chromatography-mass spectrometry using SIM acquisition mode, measuring three diagnostic ions (m/z 246, 372 and 475). Another phenothiazine derivative, fluphenazine, was used as the internal standard (I.S.). Extraction recoveries for perphenazine and I.S. were 87.6 +/- 8.2% (n=4) and 106.7 +/- 13.4% (n=4), respectively. The calibration curves were linear in the range from 4 to 100 ng/ml (r2=0.99). The limits of detection and quantification were estimated as 1.2 and 3.5 ng/ml, respectively. Precision and accuracy obtained in intra-assay studies were in the ranges of 1.3-8.7 and 1.7-19.5%, respectively, using control samples containing 6, 16 and 60 ng/ml of perphenazine. In inter-assay experiments, precision ranged from 4.3 to 14.9% and accuracy from 3.1 to 11.8%. Examples of the application of the perphenazine quantification method in otter urines after administration of perphenazine enanthate are presented. PMID- 11936698 TI - Determination of gentamicin in urine samples after inhalation by reversed-phase high-performance liquid chromatography using pre-column derivatisation with o phthalaldehyde. AB - Gentamicin and netilmicin (internal standard) were extracted from urine using C18 solid-phase extraction cartridges (94.3% recovery) and then derivatised with o phthalaldehyde and 3-mercaptopropionic acid. The derivative was stable for >6 h. The mobile phase methanol-glacial acetic acid-water (800:20:180, v/v), contained 0.02 M sodium heptanesulfonic acid, pH 3.4, and was passed at 1.0 ml min(-1) through a C18 column with fluorescence detection (excitation 340 nm, emission 418 nm). The four main components of gentamicin (C1, C1a, C2, C2a) and netilmicin, the internal standard, were separated. Using the C1a gentamicin peak, linearity was demonstrated from 0.5 to 10 microg ml(-1) and the limit of detection was 75 microg l(-1). Following 80-mg oral, 40-mg intravenous and 80-mg nebulised administration, the mean (SD) gentamicin urinary excretion was zero, 38.27 (0.96) and 1.93 (0.28) mg, respectively. Despite the relatively low lung deposition following inhalation of gentamicin the assay developed can be used to quantify the low urinary concentrations. Using this assay it should be possible to carry out urinary pharmacokinetic studies to identify the relative lung deposition of gentamicin following different methods of inhalation. PMID- 11936699 TI - Application of column-switching liquid chromatography-tandem mass spectrometry for the determination of pharmaceutical compounds in tissue samples. AB - Information on plasma-tissue distribution which is important for drug development may be obtained by "in silico" prediction tools. To support the validation of computer models, drug concentrations in rat plasma and tissues (brain, liver, kidney, testes, spleen, gut, lung. heart, muscle, skin and fat) had to be determined. In our work, we established analytical assays for a variety of substances including nicardipine, nitrendipine, felodipine and benzodiazepines. Sample preparation had to be simple and method development as well as analytical run time short to allow a high sample throughput and to minimize resources. Column-switching HPLC after homogenization and protein precipitation served as an efficient, easy and rapid sample preparation method, followed by selective MS-MS detection. Optimization of the trapping procedure was performed in order to reduce the influence of endogenous interferences and to obtain good recovery. Chromatographic separation was necessary to increase the selectivity. The use of small analytical column dimensions (2.1 x 10 mm) was investigated to achieve higher sample throughput without compromising the assay quality. Mass spectrometric parameters, such as ionization modes (positive vs. negative) and ion source types (TurbolonSpray vs. APCI) were screened to find suitable conditions for sensitive analysis of the compounds. Matrix suppression effects were taken into consideration. Calibration samples were prepared in plasma only, whereas quality control samples were prepared in both plasma and tissues to save animals and time. Accuracy and precision were in the range of 84.4-119.1% and 1 16.5%, respectively. Limits of quantification were in the range of 0.5-2.5 ng/ml for plasma and 2-10 ng/ml for tissues. Run times as short as 2.2 min could be achieved. PMID- 11936700 TI - Detection of doxorubicin and metabolites in cell extracts and in single cells by capillary electrophoresis with laser-induced fluorescence detection. AB - Capillary electrophoresis with laser-induced fluorescence detection was used to separate and detect doxorubicin and at least five metabolites from NS-1 cells that were treated with 25 microM doxorubicin for 8 h. Using 10 mM borate, 10 mM sodium dodecyl sulfate (pH 9.3) as separation buffer, the 488-nm argon-ion laser line for fluorescence excitation, and a 635 +/- 27.5 nm bandpass filter for detection, the limit of detection (S/N=3) for doxorubicin is 61 +/- 13 zmol. This low limit of detection allows for the detection of a larger number of metabolites than previously reported. Two extraction procedures were performed: a bulk liquid liquid extraction and an in-capillary single-cell lysis. While in the bulk liquid liquid extraction procedure, recovery for doxorubicin range from 50 to 99%, in single cell analysis the recovery is expected to be complete. Furthermore performing lysis of a single cell inside the separation capillary prevents doxorubicin or metabolite loss or degradation during handling. Based on the bulk method the calculated metabolite abundance is in the sub-amol per cell range while it varies from 0.1 to 1.1 fmol per cell in single cell analysis confirming metabolite loss during handling. Each metabolite was found at a level less than 0.1% of the doxorubicin content in either method, suggesting a slow metabolism in the NS-1 cell system or effective removal of metabolites by the cell. PMID- 11936701 TI - Estimating the starting dose for entry into humans: principles and practice. AB - BACKGROUND: Selection of the starting dose for the entry into humans (EIH) study is an essential first step in clinical drug development. OBJECTIVES: This paper is a review of different approaches that may be used to calculate the starting dose, presents the results of a current practice survey that reflect practice patterns at a large pharmaceutical company, and discusses selected topics related to the calculation of the starting dose. RESULTS: The methods used in the field of oncology for cytotoxic compounds are usually derived from a dose associated with some toxicity in animals multiplied by a safety factor. In therapeutic areas other than oncology, methods may be classified as four different approaches: (1) dose by factor methods that utilize the no observable adverse effect level (NOAEL) from pre-clinical toxicology studies multiplied by a safety factor; (2) the similar drug approach that may be used when clinical data are available for another compound of the same chemical class as the investigational drug; (3) the pharmacokinetically guided approach that uses systemic exposure rather than dose for the extrapolation from animal to man; and (4) the comparative approach that consists of utilizing two or more methods to estimate a starting dose and then critically comparing the results to arrive at the optimal starting dose. A "real life" example illustrates the use of each method. Advantages, limitations, and underlying assumptions of each of the methods are discussed. The results of the survey showed that the pharmacokinetically guided approach is the most commonly used method, followed by dose by factor methods. CONCLUSION: The task of estimating the starting dose is moving beyond empirical methods to those that are increasingly more systematic and theory based. PMID- 11936702 TI - Inhibitory effects of H1-antihistamines on CYP2D6- and CYP2C9-mediated drug metabolic reactions in human liver microsomes. AB - OBJECTIVE: To screen the inhibitory effects of H1-antihistamines on hepatic bufuralol 1'-hydroxylation and on tolbutamide 4-methylhydroxylation in human liver microsomes. METHODS: Bufuralol 1'-hydroxylation and tolbutamide 4 methylhydroxylation were used as index reactions for CYP2D6 and CYP2C9, respectively. The metabolites of both reactions were measured using high performance liquid chromatography and were used as indicators of whether CYP2D6 or CYP2C9 activities were inhibited or unaffected by the agents. RESULTS: All five H1-antihistamines studied showed a concentration-dependent inhibition of CYP2D6-mediated bufuralol 1'-hydroxylation with 50% inhibitory concentration (IC50) values of 32-109 microM. Cyclizine and promethazine showed inhibitory effects on tolbutamide 4-methylhydroxylation with IC20 values of 85 microM and 88 microM, respectively. Tripelennamine, chlorpheniramine, and diphenhydramine showed no inhibitory effects on CYP2C9. CONCLUSION: All five H1-antihistamines studied inhibited CYP2D6 markedly, but only cyclizine and promethazine inhibited CYP2C9 at concentrations above that usually seen in plasma. Promethazine and chlorpheniramine inhibited CYP2D6 at concentrations that are very close to their therapeutic plasma concentrations. Further studies in humans, especially in poor metabolizers of CYP2D6, will be required to confirm these findings. PMID- 11936703 TI - Paracetamol disposition in renal allograft recipients. AB - OBJECTIVE: To evaluate the disposition of paracetamol in renal allograft recipients. METHOD: Eight fasting renal allograft recipients were given 1 g soluble paracetamol orally. RESULTS: Paracetamol was absorbed rapidly, and the mean plasma half-life from 2 h to 8 h was 2.6 +/- 0.5 h. After that, disappearance of paracetamol from the plasma was slower. Both the glucuronide and sulphate conjugates of paracetamol had slow elimination half-lives of 15.1 +/- 8.8 h and 26.2 +/- 14.6 h, and there were residual amounts of both conjugates in the plasma at 24 h. The renal clearances of both conjugates were low (21 +/- 14.2 ml/min and 32.4 +/- 31.4 ml/min) and there was a significant negative correlation between total amount of paracetamol recovered in the urine in 24 h and serum creatinine (r = -0.89, P<0.01). CONCLUSION: Paracetamol disposition is abnormal in renal allograft recipients and seems to relate to abnormal renal function in these patients. PMID- 11936704 TI - N-acetyltransferase 2 polymorphism in sporadic Parkinson's disease in a Polish population. AB - OBJECTIVE: A genetic background of Parkinson's disease has been suggested, including genes implicated in xenobiotic metabolism. So far, many candidate genes responsible for the occurrence of the disease have been enumerated. This study was carried out to determine the presence of N-acetyltransferase 2 polymorphism in Parkinson's disease patients in a Polish population. METHODS: Fifty-four patients with diagnosed sporadic Parkinson's disease and 81 healthy individuals were enrolled into the study. The N-acetyltransferase 2 alleles (*4, *5, *6 and *7) were identified using polymerase chain reaction-restriction fragment length polymorphism methods with DNA extracted from peripheral blood. RESULTS: A preponderance of slow acetylators in patients with Parkinson's disease was demonstrated. Among 54 subjects with parkinsonism, 64.8% were homozygous for two mutated alleles responsible for the slow-acetylator phenotype. In the control group, a predominance of fast acetylators was noted. Subjects homozygous and heterozygous with genotypes determining fast acetylation constituting 53% of subjects, whereas 47% were slow acetylators. Comparison of the two groups of the study, i.e. Parkinson's disease and healthy individuals, revealed a statistically significant predominance of slow acetylators in Parkinson's disease patients (P < 0.05). The risk of Parkinson's disease development was more than two times greater in slow acetylators than healthy subjects. The frequency of point mutations was similar both in patients with Parkinson's disease and the healthy controls. CONCLUSION: Slow-acetylation genotype may be an important factor of individual susceptibility to Parkinson's disease. PMID- 11936705 TI - No clinically relevant drug interaction between paracetamol and phenprocoumon based on a pharmacoepidemiological cohort study in medical inpatients. AB - OBJECTIVE: A recent case control study suggests that paracetamol at a dosage above 1300 mg/day increases the anticoagulant effects of warfarin. In many European countries, phenprocoumon and not warfarin is used as an anticoagulant. However, the effects of paracetamol on the anticoagulant effects of phenprocoumon have so far not been evaluated. METHODS: This study is based on the data recorded prospectively between 1996 and 1998 in two Swiss teaching hospitals in the pharmacoepidemiological database of the SAS/CHDM project. As a first step, all phenprocoumon-treated patients with at least one international normalized ratio (INR) determination at least 4 days after cohort entry were selected and evaluated concerning paracetamol co-administration. The "paracetamol group" included only patients receiving at least 1300 mg/day paracetamol on the 3 days preceding the INR determination, whereas the comparison group contained only patients without paracetamol exposure during hospital stay. Thereafter, INR values of the paracetamol and the comparison groups were compared. RESULTS: The paracetamol and the comparison groups included 54 and 180 patients, respectively. Patients' characteristics in both groups were comparable. Patients in the paracetamol group received on average 2220 +/- 651 mg/day paracetamol. The median duration of paracetamol exposure was 5 days. The median difference in INR values between the paracetamol and the comparison groups was -0.3 (with a 99% confidence interval of -0.6, 0.1). CONCLUSIONS: These results suggest that paracetamol co administration at a dosage of 2000-2500 mg/day for 3 days has no clinically relevant effects on the anticoagulant effects of phenprocoumon. PMID- 11936707 TI - The pharmacokinetics of ketobemidone are not affected by CYP2D6 or CYP2C19 phenotype. AB - The pharmacokinetics of orally administered ketobemidone were investigated in three groups of healthy volunteers with respect to cytochrome P450 (CYP) 2D6 and CYP2C19 phenotypes: extensive metabolisers (EMs) of debrisoquine and mephenytoin (EMdeb/EMmeph), poor metabolisers (PMs) of debrisoquine (PMdeb/EMmeph) and poor metabolisers of mephenytoin (PMmeph/EMdeb). Peak plasma concentration, oral clearance, area under the plasma concentration-time curve, half-life, volume of distribution and mean residence time were not significantly different among the three groups. There was no correlation between oral clearance of ketobemidone and debrisoquine or mephenytoin metabolic ratios. Further, the urinary excretion of ketobemidone and norketobemidone was not affected by the phenotype for either CYP2D6 or CYP2C19. However, a substantial variation in plasma concentration was observed within all three groups. The results indicate that the metabolism of ketobemidone is not dependent on CYP2D6 or CYP2C19. PMs of debrisoquine or mephenytoin, as well as patients who are concomitantly treated with inhibitors of CYP2D6 or CYP2C19, are not expected to be at higher risk of adverse effects. However, due to the interindividual variability in plasma levels of ketobemidone, independent of phenotype, individual dosing based on the clinical response and therapeutic drug monitoring is recommended. PMID- 11936706 TI - Pharmacokinetics and non-analgesic effects of S- and R-ketamines in healthy volunteers with normal and reduced metabolic capacity. AB - OBJECTIVE: There is a growing interest in low-dose ketamine as an analgesic agent in different intractable pain conditions. Due to its narrow therapeutic window, well-defined pharmacokinetic parameters are essential for its successful use in these situations. Arterial data for ketamine or its enantiomers have not been reported before. The metabolic pathways involved in the metabolism of S- and R ketamines are not known. METHODS: Ten healthy male volunteers received 7 mg infusions of R- and S-ketamine-hydrochloride in a randomised order over 30 min on 2 separate days. Six were extensive metabolisers, two were poor metabolisers of debrisoquine (CYP2D6) and two were poor metabolisers of mephenytoin (CYP2C19). Arterial and venous concentrations and non-analgesic side effects were measured. RESULTS: Subjective side effects were mild but more pronounced for S- than for R ketamine. There were no salient differences between the subjects with reduced and normal metabolic capacity in pharmacokinetic parameters or in side effects. Volumes of distribution and mean residence times were 40% smaller for arterial than for venous data. The mean clearance of R-ketamine, 0.020 l min(-1) kg(-1), was slightly but significantly lower than of S-ketamine, 0.024 l min(-1) kg(-1). CONCLUSIONS: There are large differences between arterial and venous data in the pharmacokinetic parameters that are heavily dependent on distribution processes. Parameters mainly reflecting elimination, such as clearance and area under the concentration time curve, are unchanged. The choice of sampling site could be important when computer-controlled infusions are used. PMID- 11936708 TI - A pharmacokinetic and pharmacodynamic model for intravenous bisphosphonate (pamidronate) in osteoporosis. AB - BACKGROUND: Bisphosphonates are used extensively in the management of skeletal disorders including osteoporosis. Intermittent i.v. regimens with bisphosphonates are being explored as alternatives to oral regimens. The design of therapeutic regimens with bisphosphonates is mainly based on pharmacodynamic (PD) information. A combined pharmacokinetic (PK) and PD model of bisphosphonate action can help in the design of more effective therapeutic strategies. AIM: The objective of this study was to hypothesise on a PK/PD model for intermittent i.v. pamidronate in patients with osteoporosis. METHODS: Serum and urine pamidronate PKs were studied in nine patients with osteoporosis treated with pamidronate (15 mg/day via i.v. infusion) on five consecutive days. Long-term renal excretion of pamidronate was extrapolated from literature data on alendronate. The PD response (urinary hydroxyproline) was assessed in 13 patients with osteoporosis, who were given the same regimen every 3 months for 1 year. Pooled serum and urine data were fitted into a physiology-based compartment model. In order to develop a mechanistically based PK and PD model, three different Emax models were examined. RESULTS: A physiology-based three-compartment model was able to describe pamidronate PKs with a central compartment representing the serum, a second compartment representing the bone surface and a third compartment representing deep bone. The typical effect of 1 year of 3-monthly i.v. therapy was described adequately using an Emax model, in which the effect depended on both the amount of pamidronate attached to bone (second compartment) and on the amount of pamidronate buried into bone (third compartment). CONCLUSIONS: A combined PK/PD model for pamidronate is described which incorporates the specific pharmacology of bisphosphonates. The model may be used to describe long-term effects of an i.v. pamidronate regimen in osteoporotic patients. PMID- 11936709 TI - Heavy metals in traditional Indian remedies. AB - The growing popularity of traditional Indian remedies necessitates a critical evaluation of risks associated with their use. This systematic review aims at summarising all available data relating to the heavy metal content in such remedies. Computerised literature searches were carried out to identify all articles with original data on this subject. Fifteen case reports and six case series were found. Their collective results suggest that heavy metals, particularly lead, have been a regular constituent of traditional Indian remedies. This has repeatedly caused serious harm to patients taking such remedies. The incidence of heavy metal contamination is not known, but one study shows that 64% of samples collected in India contained significant amounts of lead (64% mercury, 41% arsenic and 9% cadmium). These findings should alert us to the possibility of heavy metal content in traditional Indian remedies and motivate us to consider means of protecting consumers from such risks. PMID- 11936710 TI - Quality and impact of problem-oriented drug information: a method to change clinical practice among physicians? AB - OBJECTIVES: Problem-oriented drug information is characterised by health professionals actively seeking drug information through various sources. In this study our objective was to determine the quality and impact of problem-oriented drug information among physicians. METHODS: Evaluation forms accompanying 163 written answers to physicians from a drug information centre were used to examine the quality and impact of problem-oriented drug information during the period December 1996 to June 1998. Physicians were asked whether the preliminary telephone answer was useful and, furthermore, whether the written answer was fast enough, relevant, adequately comprehensive and had valuable references. Physicians were also asked whether the answer had caused any change in their clinical practice. If yes, they were then asked to describe the actual changes. RESULTS: Of 163 evaluation forms, 117 (72%) were returned by physicians. Eighty six physicians received a preliminary telephone answer and 83 (97%) stated that this was useful. Among the physicians, 92 (79%) found that the answer was fast enough, relevant, adequately comprehensive and with valuable references, while 19 (16%) found that the answer satisfied three of these four quality criteria. Seventy-one evaluation forms stated that the answer had caused a change in clinical practice. Sixty-eight (96%) of these contained a description of the change. Thirty-five evaluation forms that stated that the answers did not cause any change in clinical practice showed the same quality score as for the total group. Thus, 28 (80%) of these satisfied four and 5 (14%) satisfied three of the quality criteria. Improved routines for and control of ongoing pharmacotherapy was the most common change in clinical practice reported by physicians. CONCLUSION: The results show that, in general, physicians found problem-oriented drug information to be of high quality, and that it had an impact on their clinical practice. Problem-oriented drug information could be a method to change clinical practice among physicians. PMID- 11936711 TI - Descriptive study and pharmacotherapeutic intervention in patients with epilepsy or Parkinson's disease at nursing homes in southern Sweden. AB - OBJECTIVES: To describe the drug use in epilepsy and Parkinson's patients living in nursing homes and to evaluate the impact of multi-speciality team intervention on health-related quality of life, activities of daily living (ADL) and confusion state. METHODS: Nursing home residents with epilepsy or Parkinson's disease in the county of Skane in Sweden were identified. From 119 nursing homes, 262 patients were identified. After obtaining informed consent, 157 patients from 48 nursing homes were included. Of these patients 74 were diagnosed with epilepsy and 84 with Parkinson's disease (one patient had both diagnoses). The average age of the epilepsy patients was 79 years and of the Parkinson's patients 81 years. Pharmacists documented the patients' drug use and any drug-related problems after communication with nursing-home residents, their contact persons at the nursing home and the residents' physicians. A multi-speciality group consisting of pharmacists, a primary care physician, a neurologist, a neuro-psychiatrist and a clinical pharmacologist evaluated the patients' medication and, when appropriate, suggested changes. Lists of each resident's medications were collected together with information about drug-related problems. The use of drugs deemed inappropriate for geriatric nursing-home residents according to Beer's criteria was documented. Health-related quality of life was evaluated using a generic health-related quality of life instrument, SF-36. Confusion state was measured using the Behaviour Pathology in Alzheimer's Disease Rating Scale (Behave-AD), and ability to perform ADL was assessed using the Schwab and England capacity for daily living scale. All measurements were repeated after approximately 6 months. During that period, for the group randomised to active intervention, the physicians involved in the care of the patients had received the recommendations for changes in drug treatment from the multi-speciality group. RESULTS: Epilepsy patients at nursing homes used on average 8.0 drugs for continuous use whereas Parkinson's patients used 8.6 drugs. According to Beer's criteria about 40% of both patient groups used drugs that are classified as inappropriate to geriatric nursing-home patients. Dopamine receptor-blocking psychotropic drugs were used by 29% of the Parkinson's patients. Indication for a patient's total drug treatment was not documented for 50% of epilepsy and 40% of Parkinson's patients. There were no significant differences between the active and control groups in changes in SF-36, Behave-AD or ADL for epilepsy patients. For Parkinson's patients there was a significant decrease in ADL for the active group, whereas there were no differences in SF-36 or Behave-AD. CONCLUSION: Nursing-home residents with epilepsy or Parkinson's disease use many drugs and often drugs that are classified as inappropriate. A simple problem-oriented questionnaire may be helpful in identifying specific drug-related problems in geriatric patients with common neurological diseases. Methods on how to improve the pharmacotherapy of these patients still have to be developed. PMID- 11936712 TI - Forged medical prescriptions in a community pharmacy network in Midi-Pyrenees area: assessment of a falsification ratio. PMID- 11936713 TI - A double-blind comparison of the bronchodilatory effect of cumulative doses of salbutamol from Diskus (Accuhaler) and Turbuhaler. PMID- 11936714 TI - Significant reduction of sertraline plasma levels by carbamazepine and phenytoin. PMID- 11936715 TI - Heparin "hitts" again. PMID- 11936716 TI - Evaluation of isolated case reports on hepatotoxicity. PMID- 11936717 TI - Predictors of posttraumatic stress reactions in Norwegian U.N. peacekeepers 7 years after service. AB - A sample of 1,624 Norwegian veterans from the UNIFIL (United Nations Interim Force in Lebanon) was investigated on average 6.6 years after service, completing a questionnaire focusing on stress exposure and posttraumatic stress reactions. The prevalence of posttraumatic stress disorder (measured by the Posttraumatic Symptom Scale [PTSS-10]) was 5% overall, but 16% in the subgroup of personnel having been prematurely repatriated from UNIFIL. Multiple regression analyses showed that the following variables made separate and significant contributions to the explained variance of the PTSS-10: Service stress exposure, perceived lack of meaningfulness with respect to the military mission, and stressful life-events in life after service. These factors explained 25% (overall sample) and 37% (repatriated sample) of the variation in the posttraumatic symptom score. PMID- 11936718 TI - After the Omagh bomb: posttraumatic stress disorder in health service staff. AB - In this postal survey of 1064 health service staff working closest to the Omagh bombing in Northern Ireland, approximately half reported having professional orcivilian involvement. Types of involvement and posttraumatic stress disorder (PTSD) levels varied between staff groups. Staff involved both professionally and as a civilian, particularly those who witnessed the trauma, or those who had experienced previous emotional problems and trauma, had the highest levels of symptomatology. Although staff with higher PTSD symptoms were more likely to seek professional help, only a minority contacted professionals for support. PMID- 11936719 TI - Preventing traumatic stress: public health approaches. AB - Population-based approaches to the primary prevention of posttraumatic stress disorder (PTSD) focus on the prevention of the stressor itself. Policy decisions also consider ways to allocate resources to best reduce potential damage from traumatic stressors and to ameliorate any resulting harm. A balance between broad risk prevention approaches and narrower treatment and recovery strategies can redistribute the risk of exposure and lead to fewer cases. Understanding that PTSD and its costs affect not only individuals who seek care, but also many others whose lives over-lap with these individuals as well as society as a whole, further informs and shapes prevention decisions. PMID- 11936720 TI - Comorbid anxiety disorders in civilians seeking treatment for posttraumatic stress disorder. AB - Research indicates that posttraumatic stress disorder (PTSD) is associated with high rates of comorbid psychiatric diagnoses. Yet, it remains unknown whether PTSD is associated with greater comorbidity relative to patients with other anxiety disorders. This study examined prevalence of comorbid anxiety disorders with PTSD relative to other disorders among a treatment-seeking population. Patients with PTSD (n = 83) evidenced greater overall comorbidity as compared to patients with other anxiety (n = 151) or Axis I (n = 73) disorders. Compared to patients with panic disorder, patients with PTSD were more likely to be diagnosed with depression and social phobia, but not other anxiety disorders. Extent of anxiety disorder comorbidity was not related to PTSD severity. These findings are discussed in terms of their relevance for treatment of PTSD. PMID- 11936721 TI - The aetiology of postpsychotic posttraumatic stress disorder following a psychotic episode. AB - This paper examines the aetiology of postpsychotic posttraumatic stress disorder (PP/PTSD) symptoms in a sample of participants who had been hospitalized following a psychotic episode. Forty two people hospitalized for a psychotic illness were interviewed during recovery to investigate whether a psychotic episode was associated with PTSD symptomatology. All participants found psychosis and hospitalization highly distressing. PP/PTSD symptoms were not associated with demographic factors, previous trauma, treatment, or insight. The PP/PTSD group reported more distress and intrusive memories associated with illness and treatment experiences and had higher scores for anxiety and dissociative symptoms. The development of PP/PTSD phenomenology was associated with the psychological distress of the experience. PMID- 11936722 TI - Peritraumatic dissociative experiences, trauma narratives, and trauma pathology. AB - Peritraumatic dissociation, i.e., dissociation during or immediately after a traumatic event, has been associated with persistence of trauma-related pathology. Peritraumatic dissociation may interfere with encoding of traumatic memories and this style may impede recovery. This study examines this hypothesis by analyzing trauma narratives from 28 female sexual and nonsexual assault victims who reported either high or low peritraumatic dissociation. Participants were asked to recount their assault. Narratives were videotaped, transcribed, and coded. Narratives of individuals with high peritraumatic dissociation had higher grade levels and a trend toward lower reading ease than those with low peritraumatic dissociation. Both higher grade levels and lower reading ease of prethreat sections of trauma narratives were related to posttreatment reexperiencing and anxiety symptoms. PMID- 11936723 TI - Children exposed to warfare: a longitudinal study. AB - Following the 1991 Gulf War a group of 94 children in Iraq were interviewed at 6 months, 1 year, and 2 years after the war. The group was exposed to the bombing of a shelter where more than 750 were killed. Selected items from different inventories, including the Impact of Event Scale (IES) assessed children's reactions. Results reveal that children continue to experience sadness and remain afraid of losing their family. Although there was no significant decline in intrusive and avoidance reactions as measured by the IES from 6 months to 1 year following the war, reactions were reduced 2 years after the war. However, the scores were still high, indicating that symptoms persist, with somewhat diminished intensity over time. PMID- 11936724 TI - Sexual harassment and PTSD: is sexual harassment diagnosable trauma? AB - Sexual harassment has become a major social, legal, and mental health problem because of its high prevalence and its negative consequences for victims. These consequences can include decreased productivity, loss of job, decreased income, and impaired psychological and physical well-being. Despite evidence from empirical studies that victims often exhibit posttraumatic stress disorder (PTSD) symptoms, some have argued that sexual harassment does not constitute legitimate trauma. We argue that many forms of sexual harassment meet the diagnostic Criteria A1 and A2 of PTSD. Finally, the DSM-IV trauma criterion is explicated, and its relationship with sexual harassment and its effects are discussed. PMID- 11936725 TI - Child and parent response to the 1993 World Trade Center bombing. AB - This study evaluated children's symptoms 3 and 9 months after the 1993 bombing of the World Trade Center, and the relationship between parent and child reactions when only the children had been in the building. Nine children who had been trapped in an elevator, 13 who had been on the observation deck, and 27 controls completed the Posttraumatic Stress Reaction Index and a Fear Inventory. Parents completed these measures about the children and comparable measures about themselves. Exposed children reported posttraumatic stress disorder (PTSD) symptoms and disaster-related fears; their parents reported experiencing PTSD symptoms. Only parents rated children's symptoms as decreasing significantly over time. Association between child symptoms and parent symptoms increased over time. Children's initial distress predicted parents' distress 9 months postdisaster. PMID- 11936726 TI - Traumatic life events and suicide risk among jail inmates: the influence of types of events, time period and significant others. AB - Relationships between traumatic life events and suicide risk were studied in two samples of jail inmates with a low (N = 216) and a high (N = 51) suicide risk. Although nonsuicidal inmates reported a high prevalence of traumatic life events, suicidal inmates reported even higher prevalence rates. Suicidal inmates reported more episodes of sexual abuse, physical maltreatment, emotional maltreatment, abandonment, and suicide attempts by significant others. They also had experienced more traumatic life events during childhood, later life, and detention. It is concluded that traumatic life events are associated with suicide risk and that such an association remains in a population with a high prevalence of traumatic life events. It is also concluded that suicide risk is dependent of the type of life event, the timing of the event, and the type of persons involved in the event. PMID- 11936727 TI - Ovarian granulosa cell tumors in childhood. AB - Granulosa cell tumors (GCT) of the ovary are prepubertal in 5% of the patients. In girls less than 20 years old, 80% of GCTs differ from those among adults. These juvenile granulosa cell tumors (JGCTs) are usually benign. GCTs belong to ovarian sex cord-stromal tumors, the more common ovarian tumors being epidermal and germinal. The etiology of GCT remains unknown. Most young children with GCT present with precorious pseudopuberty. Among adolescents GCT often causes menstrual irregularities, virilization, abdominal swelling, and pain. When JGCT is limited to the ovaries the outcome is excellent with only salpingo oophorectomy. However, more widely spread tumors are difficult to treat and cause mortality. Cisplatin-containing chemotherapy can induce remissions in adult GCTs. Estrogens and peptide hormones, i.e., inhibin, are useful in the follow-up of the patients. The authors describe 3 children with GCT and review current data on this rare tumor from molecular biology to clinical aspects. PMID- 11936728 TI - Severe iron deficiency anemia in 42 pediatric patients. AB - The objective of this study was to determine the clinical and laboratory features of children with severe iron deficiency anemia. The authors retrospectively reviewed the charts of 198 children with iron deficiency anemia to ascertain cases of severe iron deficiency anemia. Forty-two children with severe iron deficient anemia were evaluated. The median age was 21 months (range, 7-240 months) at diagnosis and 27 children were 13-24 months of age. For 35 children the major source of calories was derived from cow's milk. The median hemoglobin was 4.6 g/dL (range, 2.1-6 g/dL) and the median serum ferritin was 5 microg/L (range, 1-11 microg/L). Twenty-nine received oral iron and 13 required parked red blood cell transfusions because of co-morbid cardiorespiratory distress. Severe iron deficiency anemia mostly affects children during their second year of life. Oral iron therapy is sufficient for most children, but packed red blood cell transfusions may be needed. PMID- 11936729 TI - Chemo- and radiosensitivity testing in a patient with ataxia telangiectasia and Hodgkin disease. AB - Treatment of Hodgkin disease (HD) in ataxia telangiectasia (AT) patients is hampered by hypersensitivity to radiation and chemotherapy. Most patients die, due to toxicity or, rarely, to progressive disease. The authors report on a 9 year-old girl with stage IIA HD and AT She was treated with a tailored combined modality approach. No unacceptable toxicity was found, but the girl died of a relapse outside the irradiation field. In comparison with fibroblasts of non-AT patients, the fibroblasts of the patient were 3 times as sensitive for radiotherapy but just 1.2 times as sensitive for doxorubicin. A good correlation was shown between in vitro radio- and chemosensitivity testing and the observed clinical toxicity. The authors suggest, therefore, treating AT patients as much as possible according to standard protocols by adjusting the radiotherapy delivery and the chemotherapy regimen to individual doses derived from in vitro radio- and chemosensitivity testing. PMID- 11936730 TI - Prophylaxis with urokinase in pediatric oncology patients with central venous catheters. AB - This study evaluated the effects of urokinase in the prevention of central venous catheter (CVC)-related complications in children with malignancy. Fifteen patients with 16 CVCs (study group A) received an intraluminal application of urokinase (10,000 IU in each catheter lumen for 4 h) once a week. They were monitored prospectively with quantitative blood cultures and ultrasonography (color Doppler ultrasound of the great veins and echocardiography). The rate of complications was compared with that of 15 children with 19 CVCs without thromboprophylaxis, treated the previous significantly lower incidence of CVC dysfunction year (control group B). The authors found a wer incidence of CVC dysfunction (3/16 versus 13/19), no major thrombosis, fewer CVC-related bacteremias (2/16 versus 8/19), and a higher salvage of CVCs (1/16 versus 5/19 CVC removals due to persistent bacteremia) in the thromboprophylaxis group. Asymptomatic thrombosis rate was also lower (7/16 cases in group A versus 9/11 in group B when sonography was performed). No hemorrhagic complications were noted. Thromboprophylaxis with urokinase seems a safe and effective measure for reducing the rate of CVC-related complications. PMID- 11936732 TI - Methadone and neonatal thrombocytosis. AB - Maternal treatment with methadone sometimes induces a late-onset neonatal withdrawal in the offspring. Thrombocytoses in the newborn are very rare and sometimes are induced by methadone. T'hese newborns must be admitted for at least 2 weeks and their platelet counts must be monitored. PMID- 11936731 TI - Mucosal pathology of the upper gastrointestinal tract associated with intensive chemotherapy in children: vitamin A supplements do not prevent lesions. AB - Intensive chemotherapy (ICT) for a malignant disease in children may be associated with clinically significant mucosal lesions of the upper gastrointestinal tract. This prospective and randomized study was conducted to evaluate more thoroughly the mucosal damage and to find out whether vitamin A therapy might prevent the development of these lesions. Gastroduodenoscopy and non-invasive methods were used to examine a consecutive series of 20 patients (10 females, 10 males, aged 1-15 years) 4 weeks after initiating the therapy regimen. Half of the patients were randomized to take vitamin A supplements for 6 weeks. During a follow-up of 6 weeks, 13 (65%) reported some symptoms of the gastrointestinal (GI) tract, diarrhea and mouth pain being most prominent. Endoscopic abnormalities were found in 13 (65 %) subjects: esophagitis in 10, erosive duodenitis in 8, and gastritis in 7. Histologically, 11 subjects had duodenitis, 5 had gastritis, 3 had eosinophilic esophagitis, and 2 had lymphocytic esophagitis. Both endoscopic and histological abnormalities of the duodenum showed a close relationship with long-term (more than 2 weeks) granulocytopenia. The 4 patients with the most extensive endoscopic abnormalities were treated with HCl inhibitors, and re-endoscopy performed 4-8 weeks later showed complete recovery. The sugar permeability values, measured as the lactulose/mannitol ratio were comparable to the values obtained in the controls, and lactose intolerance was found in only 3 (20%) of the 15 children able to perform the breath test. Both the incidence of the reported GI symptoms and the endoscopically or histologically observed GI lesions were similar in the subjects randomized to take vitamin A supplements as in the controls. In this study, two thirds of children with ICT showed erosive mucosal lesions of the upper gastrointestinal tract, and vitamin A therapy failed to prevent them. Endoscopic examination is recommended if a patient has severe symptoms indicative of mucosal pathology. PMID- 11936733 TI - Paucity of hematological neoplasia after treatment of Hodgkin disease: observation after long-term follow-up at Cancer Institute, Chennai, south India. AB - The purpose of the study was to evaluate the incidence of second malignancies in childhood Hodgkin disease treated with COPP and COPP/ABV (replacing mechlorethamine by cyclophosphamide) chemotherapy. In 212 children with Hodgkin disease who were under 14 years of age and treated at Cancer Institute during the 25-year period of 1970-1994, the occurrence of second malignant neoplasms was analyzed as on 31 December 1999. Eighty-two percent attained complete response. The 5- and 10-year overall survival rate was 91 and 83%, respectively. In this interm report 5 cases of second malignancies were documented. All 5 were solid tumors: one each of soft tissue sarcoma, dermatofibrosarcoma, micropapillary carcinoma of thyroid, malignant phylloides tumor of breast, and chondrosarcoma of ilium. All patients had received combination chemotherapy and radiotherapy. Interestingly, all were splenectomized. All these patients had advanced stage of cancer and were 7-14 years of age at the time of diagnosis of first primary Hodgkin disease. It is significant that there were no secondary hematological malignancies. COPP and COPP/ABV are effective therapeutic regimens. The paucity of secondary hematological malignancies is unique in this series and may be attributed to the substitution of nitrogen mustard with cyclophosphamide in the chemotherapy combination. This is an initial observation, and further follow-up is needed for a firm conclusion. PMID- 11936734 TI - Encephalitis and myocarditis in a child with acute lymphoblastic leukemia: role of coxsackievirus B5? AB - Enteroviruses are common causes of viral encephalitis in childhood and the most common cause of myocarditis. The prognosis is good with exception of the immunocompromised children who are at higher risk with increased mortality. A case of a 7-year-old boy with acute lymphoblastic leukemia and coxsackievirus B5 associated encephalitis and myocarditis is described. The boy was in complete remission and coxsackievirus B5 infection occurred 22 months after the beginning of chemotherapy. The clinical manifestations were fever, seizures, and altered consciousness. He underwent only supportive treatment. He had an excellent outcome; 2 years later he is still in complete remission with normal electroencephalogram and normal cardiac function. PMID- 11936735 TI - Value of antimicrobials in the prevention and treatment of tuberculosis in the United States. AB - OBJECTIVE: To estimate the economic value of antimicrobials for the prevention and treatment of tuberculosis in the United States from 1954 through 1997. DESIGN: Published sources were used to estimate the burden of illness (direct medical costs, reductions in quality of life, and years of life lost) from active tuberculosis (TB) cases diagnosed between 1954 and 1997. Published literature concerning the pre-antimicrobial incidence rate and treatment of TB were extrapolated to estimate the burden of illness that would have occurred in the absence of antimicrobials. RESULTS: Between 1954 and 1997, the use of antimicrobials reduced the number of newly diagnosed cases of active TB by 32% (relative to the number that would have occurred in the absence of antimicrobials), the number of mortalities by 81%, the number of life-years lost by 87%, and the cost of medical treatment by 76%. The total financial burden of illness over this time period (including the value of lost life-years) was reduced from $894 billion (in 1997 dollars) to $128 billion. CONCLUSION: TB antimicrobials had a substantial health impact in the US from 1954 to 1997. This quantitative assessment of the economic impact of innovative biopharmaceutical products demonstrates the importance of continuing medical innovation. PMID- 11936736 TI - The nationwide tuberculosis drug resistance survey in Mongolia, 1999. AB - SETTING: Mongolia, a country in the Western Pacific Region burdened with many cases of tuberculosis, with rapid expansion of DOTS over the last several years. OBJECTIVE: To determine the prevalence of resistance to major anti-tuberculosis drugs among tuberculosis patients who have never been treated previously. DESIGN: Sputum specimens were collected from all smear-positive tuberculosis patients identified from 1 November 1998 to 1 May 1999. RESULTS: Resistance to any of the four major drugs (streptomycin, isoniazid, rifampicin, and ethambutol) was as high as 28.9% (95%CI 24.7-33.5), primarily due to high streptomycin resistance of 24.2% (95%CI 20.3-28.6). Isoniazid resistance was also high, at 15.3% (95%CI 12.1 19.1). Resistance levels to ethambutol and rifampicin were relatively low, at 1.7% (95%CI 0.8-3.5) and 1.2% (95%CI 0.5-2.9), presumably because these drugs were only recently introduced into Mongolia. Multidrug resistance was also rare, at 1.0% (95%CI 0.1-1.8). Drug resistance rates were higher in middle-aged patients than in younger and older age groups combined (P = 0.006). Males tended to have higher resistance than females, although this was of statistically marginal significance (P = 0.08). No significant regional differences in drug resistance were found. CONCLUSION: While multidrug resistance was rare, isoniazid resistance was very common, which necessitates closer monitoring of the treatment outcomes of individual patients as well as long-term follow-up for drug resistance on a nationwide scale. PMID- 11936737 TI - Tuberculosis in Spain: epidemiological pattern and clinical practice. AB - SETTING: Thirteen Autonomous Regions in Spain. OBJECTIVE: To study the incidence of all forms of tuberculosis (TB) and investigate clinical practice in TB. DESIGN: Cases of all forms of tuberculosis diagnosed in the study setting from May 1986 to April 1997 were identified though active search of different databases. Clinical and epidemiological information on cases was collected from clinical records or by interview with physicians. RESULTS: The overall incidence of all forms of tuberculosis was 38.5/100,000 and the incidence of smear-positive disease was 13.83/100,000. Most cases (97.1%) were Spanish nationals, with rates higher in men than in women (52.7/100,000 vs. 24.87/100,000) and in groups aged 25-34 and 75 years and over (61.35/100,000 and 59.35/100,000, respectively). Disseminated forms were frequent (6.6%), and the most common risk factor was human immunodeficiency virus (HIV) infection (17.7% of cases). Hospitalisation was common (71.6%). Microbiological confirmation of diagnosis was sought for 87.7% of the cases (91.8% of pulmonary vs. 75.5% of extra-pulmonary cases), and 65.2% were culture-positive (73.8% of pulmonary vs. 39.7% of extra-pulmonary cases). HIV-infected patients were treated in almost equal proportions with three or four drugs (49.7% and 48.2%, respectively), while HIV-negative cases or those whose HIV status was unknown were usually treated with three drugs. CONCLUSION: The epidemiological pattern of TB in Spain is different to other industrialised countries in the age distribution of cases and the proportions of foreigners and cases with HIV infection. Microbiological confirmation of diagnosis is more common in pulmonary than in extra-pulmonary disease, and treatment with four drugs more frequent in HIV-positive cases. PMID- 11936738 TI - Socio-economic determinants of knowledge and attitudes about tuberculosis among the general population of Metro Manila, Philippines. AB - SETTING: Five urban communities in Metro Manila with different socio-economic status. OBJECTIVE: To investigate general knowledge about tuberculosis (TB) and intended health-seeking behaviour for TB symptoms among the general population of Metro Manila, and how they are determined by socioeconomic factors. DESIGN AND METHOD: Subjects aged 18 years and over were interviewed in a cross-sectional survey. Data on TB knowledge and intended health-seeking behaviour were stratified by sex, age, educational background, occupation, average family income and place of residence. A TB knowledge score was constructed based on 10 selected questions. Multivariate analysis with standard logistic regression showed those factors that were independently correlated to understanding of the disease, and to attitudes in case of TB symptoms. The TB knowledge score was also analysed by TB information source. RESULTS: Completed surveys were obtained from 3970 subjects. They scored low in general knowledge about TB (3.64 +/- 1.67, range 0 10). A higher score was independently associated with college education (OR = 4.35, 95%CI 6.60-2.87, P < 0.0001). There was no significant difference in TB knowledge acquired through several sources. No intention of seeking health care and intention to self-treat in case of TB disease were significantly correlated to low family monthly income (OR = 7.10, 95%CI 8.25-6.11, P < 0.0001 and OR = 1.74, 95%CI 2.06-1.46, P < 0.0001, respectively). CONCLUSION: Socio-economic factors should be taken into account in the design of TB information campaigns and in prioritising public health interventions about TB. PMID- 11936739 TI - Factors affecting patient compliance with anti-tuberculosis chemotherapy using the directly observed treatment, short-course strategy (DOTS). AB - SETTING: Kota Kinabalu and surrounding communities in Sabah, Malaysia. OBJECTIVES: To establish factors affecting compliance of patients with anti tuberculosis chemotherapy, their knowledge of the disease, and views on improving the DOTS strategy. DESIGN: Interviews with compliant patients attending clinics for DOTS treatment and with non-compliant patients in their homes, in August and September 2000. RESULTS: A total of 63 compliant and 23 non-compliant patients were interviewed. For non-compliant patients, reaching the treatment centre entailed greater cost (P < 0.005) and travel time (P < 0.005) compared to compliant patients. Cost of transport was the reason most frequently given for non-attendance. Non-compliant patients were more likely to have completed secondary education (P < 0.05), and to be working (P < 0.01). More non-compliant patients had family members who had had the disease (P < 0.01). There was no difference between the groups for overall tuberculosis knowledge scores; however, non-compliant patients were more likely to think that treatment could be stopped once they were symptom free (P < 0.01). Most patients (73%) felt that the DOTS system could be improved by provision of more information about tuberculosis. CONCLUSION: Compliance with DOTS in the Kota Kinabalu area is affected by travel expenses, time spent travelling to treatment centres, and having family members who have had the disease. Patients would like more information on tuberculosis. PMID- 11936740 TI - Managing medication compliance of tuberculosis patients in Haiti with medication monitors. AB - SETTING: A tuberculosis clinic in Haiti using self-administered medication. OBJECTIVE: To determine if medication monitors could be used along with directly observed therapy in developing countries to help solve the problem of compliance with medication. DESIGN: Patients were randomized into three groups: Group A took medication from medication monitors and were given counseling based on the monitor record; Group B took medication from medication monitors, but the record was not available for counseling; and Group C took medication from simple containers. RESULTS: Good monitor records in the first 11 weeks predicted less default from treatment (P < 0.01), and better compliance (P < 0.01) in the last 9 months. Counseling based on the monitor record appeared to reduce treatment abandonment by about half. CONCLUSION: This study suggests that medication monitoring of self administered treatment would be useful in settings where directly observed therapy cannot be delivered for the entire duration of treatment, especially in rural areas, by 1) identifying poor compliance early in therapy, 2) reducing the frequency of clinic visits for patients with good monitor records who live excessive distances from the clinic, 3) counseling patients about their monitor record to improve treatment completion rates, and 4) lengthening the duration of therapy when poor compliance is found. PMID- 11936741 TI - Treatment outcome of relapse and defaulter pulmonary tuberculosis patients. AB - SETTING: Tuberculosis clinic in a referral hospital. OBJECTIVE: To evaluate the effect of risk factors on the outcome of retreatment in relapse and defaulter pulmonary tuberculosis patients. DESIGN: A total of 57 retreatment tuberculosis patients treated and monitored in our centre between January 1997 and June 1999 were evaluated with respect to treatment outcome. Factors which have on effect on treatment outcome were investigated. RESULTS: Of 57 patients, 37 (64.9%) were classified as relapse cases and 20 (35.1%) as defaulters. The treatment success rate was 71.9% (68.4% cure rate and 3.5% completion rate). Failure was encountered in 22.8%. Twenty-six patients (45.6%) exhibited resistance to at least one drug, namely rifampicin. The multidrug-resistance (MDR) rate was 18.5%. Treatment success rates were 100% and 50%, respectively, in patients in whom susceptibility to all drugs and resistance to at least one drug were detected. Successful outcome was possible in 68.8% of patients with any rifampicin resistance and in 20% of patients with MDR tuberculosis. Retreatment resulted in failure in 80% and 100%, respectively, of patients whose sputum cultures remained positive at the end of the second and third months. CONCLUSION: Drug resistance proved the most important factor affecting treatment outcome. Success rates in retreatment of patients with any rifampicin resistance or MDR tuberculosis are low. Conversion to negative sputum results at the end of the second and third months of retreatment seems to be a significant parameter for a successful outcome. PMID- 11936742 TI - Operational assessment of isoniazid prophylaxis in a community AIDS service organisation in Uganda. AB - SETTING: Isoniazid therapy was shown to be 70% effective at preventing tuberculosis in HIV-infected, PPD-positive Ugandan adults, but the feasibility of implementation outside an efficacy trial has not been established. OBJECTIVE: To study uptake, adherence and feasibility of a 6-month course of isoniazid preventive therapy in community-based HIV clinics in Uganda. DESIGN: Observational cohort study describing selection of patients and adherence to isoniazid 300 mg daily. Adherence was measured by clinic attendance, pill counts and urine isoniazid metabolite testing. Implementation was costed on a service delivery basis. RESULTS: Of 1597 cohort members, 22% were PPD-positive. Over 18 months, 193 PPD-positive individuals were assessed for prophylaxis and 98 (51%) were enrolled. Of those enrolled, 74 (76%) completed their course of isoniazid therapy, and 80% were fully adherent. Symptoms or previous treatment for tuberculosis and suspicion of tuberculous lymphadenopathy were the main reasons for exclusion. The additional cost of providing this service was US $14,549. CONCLUSION: Clinics specialising in the care of persons with HIV/AIDS can successfully implement isoniazid prophylaxis. Difficulties in excluding active tuberculosis and the costs of running the programme may limit its widespread implementation. PMID- 11936743 TI - The emergency department is a determinant point of contact of tuberculosis patients prior to diagnosis. AB - SETTING: Metropolitan Edmonton, Canada. OBJECTIVES: To determine 1) the pre diagnosis emergency department utilization history of urban tuberculosis patients, and 2) the resource and outcome implications of emergency department utilization by tuberculosis patients pre-diagnosis. DESIGN: Nested case (emergency department attendee) control (non-emergency department attendee) study of a retrospective cohort of tuberculosis patients. PATIENTS: All tuberculosis notifications, 1994 through 1998. MAIN OUTCOME MEASURES: Emergency department utilization during the 6 months antedating the diagnosis and emergency department attendee characteristics; for those notified in 1997 and 1998, hospitalizations, nosocomial infectiousness time, and health care costs. RESULTS: Of 250 cases of tuberculosis, 117 (47%) made a total of 258 pre-diagnosis emergency department visits. Emergency department use increased the nearer the patient was to diagnosis. Emergency department attendees were more likely to be older, to have smear and/or culture positive respiratory disease, to have a risk factor for progression of infection to disease, and to have a fatal outcome. In 1997 and 1998, emergency department throughput accounted for 70% of all hospitalization days, 95% of all source case nosocomial infectiousness time, and most health care costs of tuberculosis patients pre-diagnosis. CONCLUSIONS: The emergency department is heavily utilized by urban tuberculosis patients pre-diagnosis. Emergency department throughput of tuberculosis patients pre-diagnosis has major resource and outcome implications. The emergency department may present an opportunity for earlier diagnosis. PMID- 11936744 TI - Use of a standardised checklist to assess peripheral sputum smear microscopy laboratories for tuberculosis diagnosis in Uganda. AB - SETTING: TB diagnostic units in Uganda. OBJECTIVES: To assess and improve the supervision and performance of sputum smear microscopy in the peripheral diagnostic units in Uganda using a standardised laboratory checklist. DESIGN: A standardised checklist was developed and used during the quarterly supervisory visits of the District TB and Leprosy Supervisors for five quarters from the fourth quarter of 1997 until the last quarter of 1998. Individual peripheral laboratory performance was monitored during the study period. RESULTS: Forty eight of 304 TB diagnostic units in six of the 45 Ugandan Districts were supervised using the checklist. A total of 208 checklists were analysed. The situational analysis of the peripheral diagnostic units at the beginning and at the end of the study showed a marked improvement in laboratory performance in all aspects related to sputum smear microscopy. Individual laboratory performance was monitored over five quarters, and timely response to shortcomings was provided. CONCLUSION: The systematic use of a standardised laboratory checklist can be considered an important step forward in improving the performance of the peripheral laboratories in Uganda through on-the-spot correction of any identified shortcomings. PMID- 11936745 TI - Combined bronchoalveolar lavage and polymerase chain reaction in the diagnosis of pulmonary tuberculosis in smear-negative patients. AB - SETTING: The polymerase chain reaction (PCR) may be sensitive and specific for the diagnosis of tuberculosis, but most reports are of studies conducted in well controlled laboratories. A study to evaluate the clinical value of bronchoalveolar lavage (BAL) combined with PCR was necessary. OBJECTIVE: One hundred and thirty one patients were recruited into the study from March 1994 to February 1997. DESIGN: Patients with a positive acid-fast stain on sputum smear were recruited into group A as positive controls, patients with lung cancer and a negative acid-fast stain on sputum smear were put into group B as negative controls, and patients who had clinical symptoms of pulmonary TB without sputum or with negative smear results were the investigating group. PCR was performed on the sputum samples from group A and B patients and on the BAL fluid from those in group C. RESULTS: The sensitivity of PCR was 96% in group A, and the specificity was 100% in group B. The sensitivity of PCR in the BAL fluid from the group C patients was 36% and the specificity was 96%; the positive predictive value was 94% and the negative predictive value was 45%. CONCLUSION: BAL plus PCR is useful in the rapid diagnosis of pulmonary TB in non-productive or smear-negative patients. PMID- 11936746 TI - Widespread distribution of a single drug rifampicin formulation of inferior bioavailability in South Africa. AB - A pharmacokinetic study of rifampicin, isoniazid, pyrazinamide and ethambutol in 118 tuberculosis patients revealed low and variable concentrations of rifampicin after 2 months of treatment on standard daily doses. A group of 53 patients exposed to specific batches of formulations containing rifampicin alone showed particularly low and variable levels of the drug. The national drug regulatory authority subsequently withdrew the batches in question, as sufficient bioavailability data had not been submitted after what the manufacturer had considered to be a minor formulation change. The evidence supports initiatives to implement bioavailability testing of new formulations (and of established formulations subsequent to changes in the manufacturing process) prior to distribution. Concerns about the bioavailability of rifampicin-containing products, including those with adequate dissolution profiles, should not be confined to fixed-dose combination anti-tuberculosis drugs, but should also be applied to single drug formulations. PMID- 11936748 TI - Health seeking behaviour among tuberculosis patients in rural South Africa. PMID- 11936747 TI - Screening childhood contacts of patients with smear-positive pulmonary tuberculosis in Malawi. AB - A survey was conducted by the National Tuberculosis Programme in 44 Malawian hospitals on screening young children aged 5 years or less in households of adults diagnosed with smear-positive pulmonary tuberculosis. Of 659 hospitalised adult patients, 267 (41%) had a total of 365 young children; 56 (21%) adult patients had been informed about childhood screening, and in 31 (12%) some of their children were screened. Of the 365 young children, 33 (9%) were screened for TB--23 received isoniazid preventive therapy, six received anti-tuberculosis treatment, and in four no action was taken. Steps are needed to improve childhood screening procedures. PMID- 11936749 TI - Appropriate definition of hypoxaemia needed to determine its clinical correlates. PMID- 11936751 TI - Importance of metalloproteinases and macrophages in viper snake envenomation induced local inflammation. AB - The inflammatory action of jararhagin, a hemorrhagic metalloproteinase from Bothrops jararaca venom, was studied in mice using dorsal air pouches. The injection of the toxin in 6-day-old air pouches resulted in a leukocyte accumulation comparable to that induced by LPS and whole venom. Polymorphonuclear and mononuclear cells were present in this infiltrate, with a predominance of neutrophils. Treatment of jararhagin with 1,10-phenantroline abolished its proteolytic activity and reduced the pro-inflammatory effect in approximately 50%. Cell influx was not observed when jararhagin was injected into 1-hr air pouches devoid of macrophages, except when it was injected together with 10(6) syngeneic peritoneal macrophages. Supernatants of macrophages stimulated in vitro with jararhagin did not induce leukocyte influx in 1-hr air pouches; the influx occurred after injection of the pellets of stimulated cultures. In summary, jararhagin is an important pro-inflammatory component of B. jararaca venom, and its activity is dependent upon the proteolytic activity and the presence of macrophages. PMID- 11936750 TI - Polynitroxylated starch/TPL attenuates cachexia and increased epithelial permeability associated with TNBS colitis. AB - Free radicals play an important role in the initiation and progression of inflammatory bowel disease (IBD). Therefore, the reduction or elimination of adverse oxidant effects can provide novel therapy for IBD. Here, the antioxidant capacity and protective effects of a new class of chemically modified hetastarch (polynitroxyl starch, or PNS) plus 4-hydroxyl-2,2,6,6-tetramethylpiperidine-N oxyl (Tempol or TPL) (PNS/TPL) were assessed in a model of colitis. The superoxide scavenging capacity of PNS/TPL-that is, the inhibition of the reduction of cytochrome c in the presence of xanthine/xanthine oxidase (X/XO)-was evaluated in vitro. The effects of PNS/TPL on X/XO-induced neutrophil endothelial adhesion in vitro were investigated. Also, this study tested the protection produced by PNS/TPL in a mouse model of trinitrobenzene sulfonic acid (TNBS) induced colitis. PNS/TPL was given intravenously immediately before (< 30 min) and intraperitoneally at 24 and 72 hr after TNBS induction. The body weight and survival rate of the mice were checked daily. Colonic mucosal damage was assessed on the 7th day by measuring intestinal permeability to Evans blue (EB) in vivo. The ability of PNS to reoxidize bioreduced TPL was documented by whole-body electron paramagnetic resonance (EPR) detection. We found that PNS or TPL exhibits superoxide dismutase (SOD)-like activity, with approximately 2% of SOD activity occurring on a molar basis. The endothelial-neutrophil adherence induced by X/XO was significantly inhibited by PNS/TPL but not by TPL alone. PNS/TPL protected against cachexia and mortality, both usually induced by TNBS. Epithelial permeability was increased significantly in TNBS mice but was ameliorated by the administration of PNS/TPL. In conclusion, PNS/TPL may be beneficial in the treatment or prevention of IBD through its antioxidant effects, which inhibit oxidant-mediated leukocyte adhesion and injury to endothelial cells. PMID- 11936752 TI - Adenosine A2A receptor agonists promote more rapid wound healing than recombinant human platelet-derived growth factor (Becaplermin gel). AB - Animal studies of the topical application of adenosine A2A receptor agonists show that it promotes wound closure. To further confirm the efficacy of adenosine A2A receptor agonists as promoters of wound healing, we compared the effect of MRE0094, a novel selective adenosine A2A receptor agonist, to CGS-21680, a reference selective adenosine A2A receptor agonist, as well as to recombinant human platelet-derived growth factor (0.01% Becaplermin gel), an agent currently used to promote healing of diabetic ulcers, on wound closure in healthy BALB/C mice. Wounds (approximately 12 mm diameter) were created on the dorsum of mice (two per mouse) and then treated daily with vehicle, 0.01% Becaplermin gel, or different doses of the adenosine A2A receptor agonists. The wound margins were traced onto plastic sheets, and the wound areas were digitized, quantitated, and compared. We found that application of MRE0094 (1 microg/wound and 10 microg/wound) and CGS-21680 (1 microg/wound and 5 microg/wound) achieved 50% wound closure significantly more rapidly than control application (day 1.9, 1.9, 3.5, 3.2, respectively, versus control day 4, p < 0.05 ANOVA). Surprisingly, neither higher nor lower concentrations of CGS-21680 affected the rate of wound closure, as compared to control. In contrast, Becaplermin gel did not increase the rate at which wounds closed (50% closure by day 7.2, p = NS versus control). These data confirm our prior observations that adenosine A2A receptor agonists promote wound closure, and they suggest that these agents may be as effective if not more effective than Becaplermin gel for the treatment of poorly healing wounds. PMID- 11936753 TI - No causal association between inflammation and Chlamydia pneumoniae in patients with chronic ischemic arterial disease. AB - The C-reactive protein, Chlamydia-specific IgG antibody, and fibrinogen were assayed in the serum of 159 patients with arterial disease (the arterial group) and 203 patients with heart valve prostheses (the valvular group) and no demonstrable coronary disease. In the arterial group, the Chlamydia pneumoniae antibody was > or = 1:32 for 67.3% (107/159) of the patients, the C-reactive protein was elevated in 41.5% (66/159), and the fibrinogen was elevated in 27.7% (44/159). In the valvular group, the C. pneumoniae antibody was > or = 1:32 for 59.1% (120/203) of the patients; the C-reactive protein was elevated in 34.0% (69/203), and the fibrinogen was elevated in 17.2% (35/203). Of 107 patients in the arterial group with C. pneumoniae titers > or = 1:32, only 26 (24.3%) had elevated fibrinogen (426 +/- 29 mg/dL) and 44 (41.1%) had elevated C-reactive protein (1.06 +/- 0.52 mg/dL). Similarly, of the 120 patients in the valvular group with C. pneumoniae titers > or = 1:32, 17 (14.2%) had elevated fibrinogen (409 +/- 29 mg/dL) and 34 had elevated C-reactive protein (0.99 +/- 1.1 mg/dL). Correlated poorly was C. pneumoniae with C-reactive protein and fibrinogen levels. Only the fibrinogen level could be discriminated between the arterial and the valvular group. These results suggest that no causal association exists between inflammation and C. pneumoniae. A highly significant correlation between C-reactive protein and fibrinogen levels was found. PMID- 11936754 TI - Inhibition of tyrosine kinases blocks adhesion-induced T-cell coactivation without interfering with T-cell adhesion to endothelial cell-surface ligands. AB - Integrin and cell adhesion molecule-regulated cellular adhesion plays an integral part in the recruitment and activation of lymphocytes. T-cell activation is a dynamic process subject to integrin-dependent and -independent regulation. Stimulation of human peripheral blood T cells by the anti-CD3 monoclonal antibody results in a rapid upregulation of integrin affinity. In conjunction with adhesion to endothelial cell-derived ligands and extracellular matrix proteins, anti-CD3 antibodies have been shown to result in significant increases in IL-2 production and T-cell proliferation. Therefore, at least two signal cascades are activated by ligation of the TCR: One results in a change in affinity of integrins for their ligands, whereas the other activates a signaling cascade that leads to gene induction. We investigated the effects of several tyrosine kinase inhibitors on human peripheral blood T-cell adhesion and adhesion-induced costimulation of IL-2 expression and secretion. These compounds did not inhibit anti-CD3-induced short-term (30 min) or long-term (18 hr) T-cell adhesion to VCAM 1, MAdCAM, or ICAM-1. When T cells were stimulated with anti-CD3 and allowed to adhere to VCAM-1, MAdCAM, or ICAM-1 in the presence of these inhibitors; IL-2 production was significantly reduced. The MEK specific inhibitor, PD98059, did not block T-cell adhesion to the various substrates, but it did block IL-2 synthesis. In addition, the tyrosine kinase inhibitors and PD98059 blocked anti CD3-mediated stimulation of IL-2 synthesis. These data suggest that the signaling mechanism for anti-CD3-mediated integrin activation is distinct from the signaling pathway that results in adhesion-induced IL-2 synthesis via specific integrins and anti-CD3. PMID- 11936755 TI - A new assay to monitor the degranulation process in phagocytizing human neutrophils. AB - oxidation, we set up a method for measuring MPO intraphagosomal release in human neutrophils. The method is based on the passive engulfment of DAB together with the phagocytosable particle. Inside the vacuole, this substrate is oxidized by MPO released from the azurophilic granules. The colorimetrical evaluation of the amount of DAB oxidized allows for cheap, rapid quantification of MPO intraphagosomal secretion in whole cells. Using this method, we show that the degranulation process, involving azurophilic granules, can be monitored carefully during phagocytosis. It takes place after the ingestion of zymosan particles opsonized with normal human serum, as well as during IgG-mediated phagocytosis and under conditions where beta2 integrins are blocked. However our findings also show that the extent of intraphagosomal secretion depends on either the extent of opsonization or the type of receptor engaged during the phagocytic event. PMID- 11936756 TI - The use of traditional and Western medicine among Korean American elderly. AB - With the current wave of Asian immigration to the United States, awareness of their use of traditional medicine, often in combination with Western medicine, is an increasingly salient issue for health care professionals. This paper describes the use of traditional and Western medicine by Korean American elderly and factors that are associated with their health-seeking behaviors and health service utilization. Data were obtained from a convenience sample of 205 Korean elderly immigrants through interview. The findings revealed that health service utilization among Korean American elderly covered a broad spectrum of health resources, including the use of Western medicine, the traditional Korean medicine (hanbang), or both Western and traditional clinics. In addition, health insurance status and the source of health care were found to relate to the types of health service that Korean American elderly chose to utilize. Better understanding of patterns of health service utilization may improve the quality of health care by increasing health providers' cultural sensitivity and facilitating adequate communication between Asian immigrant clients and their care providers. Implications for research and practice are discussed. PMID- 11936757 TI - Nutritional risk needs assessment of community-living seniors: prevalence of nutrition problems and priorities for action. AB - A random mailed survey was sent to 425 members of a seniors recreation center in Guelph, Ontario for the purpose of identifying the prevalence of nutritional risk, specific nutrition problems, and educational format preferences of these seniors. This needs assessment was used to plan an education initiative called "Action Nutrition." SCREEN, a 15-item nutritional risk screening instrument, demographic data, participation in current recreation programs, and frequency of visits to the center were included in this survey. The response rate was 62%, average age was 72 years and 59% were female. Respondents attended on average one activity per month, 60% visited the center at least once per month and 79% read the monthly newsletter. 56.7% of respondents were considered to be at nutritional risk using SCREEN and common problems were: significant weight change in 6 months; low intake of fruits and vegetables and milk products; and restriction of the diet due to health reasons. This survey determined that nutrition problems and risk are prevalent among community-living seniors, justifying the development and implementation of educational initiatives in this setting. PMID- 11936758 TI - Pharmacists' perceptions of Healthy People goals in economically stressed cities. AB - The objectives of the study were: (a) to determine pharmacists' perceptions of the importance of Healthy People (HP) 2000 objectives for their patients and community; and (b) to analyze pharmacists' perceived perceptions about recent trends in the characteristics of patients. Three hundred one useful questionnaire were collected from pharmacists who either own or manage a pharmacy located in New Jersey's 67 cities eligible for financial assistance. Pharmacists consider health objectives which are directly linked to health care such as preventing, detecting, and controlling hypertension, heart disease, stroke, cancer, diabetes, and disabling conditions, to be very important. Pharmacists agree to some extent that their patients are now more willing to accept pharmacy counseling services with their prescriptions than before. In conclusion, pharmacists considered objectives related to preventive services to be the most important for HP 2000 objectives, and health promotion and health protection to be less important. The findings of this study will enhance the understanding of pharmacists' perceptions of community health issues and will be beneficial to attain the objectives of HP 2010. PMID- 11936759 TI - Recruitment and retention of physicians for primary care research. AB - The primary objective of this report is to examine factors associated with recruitment of physicians in community-based primary care research. Reported results are based on an observational study of physician recruitment efforts undertaken in a randomized controlled trial designed to improve primary care physicians' cancer screening and counseling activities. The Partners for Prevention project was a statewide randomized controlled trial of primary care physicians selected from the state of Colorado. Two-hundred and ten eligible internal medicine and family medicine practices in both rural and urban community settings of the state of Colorado were selected into this study and a sentinel physician was chosen to represent each practice. Only 6% (13/210) of recruited practices initially declined to participate in the study, but the total refusal rate had reached 30% (59/210) by the time the intervention was implemented five months later. Study participants (n = 136) were younger (mean age 45.7 vs. 50.0, p = 0.008) and more often located in a rural area (46% vs. 31%, p = 0.04) than decliners (n = 59), but there was no association with gender of the physician (87% for females vs. 95% for males, p = 0.13). Participants were more often family practice physicians by training rather than internists (75% vs. 56%, p = 0.008), whereas there was no difference in participation rates by practice type (solo versus group, 60% vs. 64%, p = 0.52). Differences in demographic, geographic, and training characteristics between trial participants and decliners suggest the potential for better targeting of recruitment efforts. Viable strategies for recruiting community-based primary care practices to research studies are proposed. PMID- 11936760 TI - The prevalence of arthritis and activity limitation and their predictors in Missouri. AB - Arthritis and other rheumatic conditions comprise the leading cause of disability in the United States. In 1990, an estimated 16.7% of Missourians had arthritis. By 2020, an estimated 20% of Missourians will have this condition. We examined Missouri's prevalence of self-reported physician-diagnosed arthritis, chronic joint symptoms, and activity limitation due to joint symptoms and their associations with selected predictors (i.e., socio-demographic, access to health care, risk factor, and comorbidity indicators) from Missouri's 1996 Behavioral Risk Factor Surveillance System. We conducted logistic regression analysis to generate Odds Ratios and 95% Confidence Intervals of arthritis and activity limitation across levels of predictors. Analysis indicates arthritis is under diagnosed in younger individuals and that arthritis and activity limitation due to joint symptoms are significant contributors to functional limitation, enhancing dependency while decreasing the quality of life. As the population ages, arthritis, chronic joint symptoms, and activity limitation will become a larger public health problem. PMID- 11936761 TI - On-column enzyme-catalyzed microreactions using capillary electrophoresis: quantitative studies. AB - Capillary electrophoresis and on-column enzyme-catalyzed microreactor techniques were used to quantitate the reaction projects resulting from three model systems: i) the conversion of nicotinamide adenine dinucleotide (NAD) to nicotinamide adenine dinucleotide, reduced form (NADH) in the oxidation of glucose-6-phosphate (glc-6-p) to 6-phosphogluconate by glucose-6-phosphate dehydrogenase (G6PDH, EC 1.1.1.49); ii) the conversion of adenosine triphosphate (ATP) to adenosine diphosphate (ADP) and adenosine monophosphate (AMP) by hexokinase (HK, EC 2.7.1.1) and apyrase (APY, EC 3.6.1.5), respectively, in the conversion of glucose to glucose-6-phosphate and inorganic phosphate, respectively, and; iii) the conversion of fructose-1,6-bisphosphate to dihydroxyacetone phosphate and glyceraldehyde-3-phosphate by fructose-biphosphate aldolase (ALD, EC 4.1.2.13). Single and double microreactor techniques employing direct or indirect detection were used to follow the conversion of substrate to product(s). In addition, electrophoresis conditions including voltage, enzyme concentration, and mixing time of the reaction, were correlated to product distribution profiles. PMID- 11936762 TI - Microemulsion electrokinetic chromatography. AB - Microemulsion electrokinetic chromatography (MEEKC) is a CE technique capable of resolving complex mixtures of charged and neutral compounds covering a wide range of solubilities. It is a relatively recently developed technique and the recent increased attention has led to a number of application reports. The background to the technique and a brief overview of reported applications is provided. PMID- 11936763 TI - Comparison of high-performance capillary electrophoresis and liquid chromatography with ultraviolet detection for the isomers of o-, m-, and p diphenol. AB - The separation conditions and determination of isomers of o-, m-, and p-diphenol in wastewater by high-performance capillary electrophoresis (HPCE) and HPLC with ultraviolet detection at 280 nm have been studied systematically. The study showed that both methods gave comparable results in retention time, linear range, recovery, reproducibility, and the amount of diphenols in real samples. PMID- 11936764 TI - Enzymatic amplification by substrate recycling for alpha-ketoglutarate in conjunction with capillary electrophoresis. AB - The application of substrate cycling for alpha-ketoglutarate (AKG) through enzymatic amplification has been demonstrated in conjunction with capillary electrophoresis. AKG is determined using an off-line assay, which involves the coupled enzymatic reactions catalyzed by glutamate dehydrogenase and glutamic oxalacetic transaminase. Varying concentrations of the substrate and set concentrations of cofactors and enzyme are incubated together outside the CE instrument. The accumulated product, nicotinamide adenine dinucleotide, reduced form (NADH), was detected at 340 nm using CE. Amplification of a factor of 10 over just the reversed glutamate dehydrogenase reactions permitted an AKG detection limit of 10 microM. Throughput for each assay is about 5 min (including wash and equilibrium steps). After simple dilution as the only sample pretreatment, spiked AKG serum samples were determined with an average recovery of 96% with a 3% RSD. PMID- 11936765 TI - The use of cyclodextrin-modified capillary electrophoresis in the determination of the enantiomeric purity. AB - Employing cyclodextrin-modified capillary electrophoresis, the racemates of some 30 acidic, neutral and basic dihydropyridines (DHP) were resolved using neutral and negatively charged cyclodextrins (CDs) derivatives. Whereas the enantiomers of the acidic DHPs could be separated with neutral CDs the enantiomers of the neutral DHPs were only baseline separated using the sulfobutylether substituted beta-CD. Working in reversed polarity mode--detection at the anode--improved both the peak shape and the resolution of the enantiomers. The racemates of the DHPs characterized by a secondary or tertiary amine function in the carboxylate side chain could be resolved by using either the neutral gamma-CD or negative charged CDs. The poor peak shape found with anionic CDs could be improved by addition of methanol. PMID- 11936766 TI - Zeta potential of staphylococcal Protein A (Z-domain) used for determination of complex constant with immunoglobulin Fc fragment by affinity electrophoresis in capillary format. PMID- 11936768 TI - A rural service-learning model for health administration education. AB - Service learning for students in the health professions can best be described as an educational methodology based on the collaboration between an academic discipline and a community agency to link student learning with community-based service. This paper describes a service-learning model in rural and medically underserved communities used in a master in health administration program in a rural Southern state. It presents the process, components, outcomes, and challenges. Over the past five years, 56 students completed 116 experiences involving internships, field projects, or special projects in rural and medically underserved communities. A total of 27 percent of our graduates who completed rural and medically underserved experiences accepted job positions in rural health care entities. Strategies utilized by this rural service-learning model include interdisciplinary team training, partnerships among rural and medically underserved community health care entities and academic institutions, faculty preceptor retreats, and involvement in local community initiatives. These experiences prepare future health administrators to contribute in a meaningful way toward building an effective rural health care delivery system. PMID- 11936767 TI - Establishment of an interaction model of human apolipoprotein H with lipid monolayer by capillary SDS gel electrophoresis. AB - Apolipoprotein H (ApoH) is a plasma glycoprotein isolated from human serum. It plays a key role in the interaction with lipids. For the first time, the concentration of ApoH adsorbed on the lipid monolayer has been determined, and was done so using capillary electrophoresis. Based on this determination, an interaction model of ApoH and lipid monolayer was constructed, and this interaction is one of nonspecific adsorption. A neutral coated capillary (50 cm x 100 microm i.d.) and a negative voltage of 15 kV were used to separate ApoH. The calibration curve of ApoH was built using a detection limit of 50 microg/mL(-1) (near to 1 microM), and the RSD of the relative migration time of ApoH was 1.4% (n = 7). PMID- 11936769 TI - Benchmarking graduate health administration programs: stakeholders' perceptions. AB - The purpose of this study was to identify and compare among key stakeholders the factors in graduate health care administration education that are perceived to be important for ranking, or benchmarking, based on the opinions of those stakeholders, i.e., program directors, faculty, graduate students, and accrediting agency commissioners. We used an original survey to obtain stakeholders' perceptions and opinions about important process and outcome measures. We sent it to all ACEHSA-accredited graduate health care administration programs in the United States, Canada, and Puerto Rico; to full-time faculty members in each program; to three current graduate students in each program, and to all ACEHSA commissioners. We performed frequency of responses, Analysis of Variances (ANOVA) tests, and Dunnett T3 tests. A response rate of 32 percent (n = 156) was achieved for all stakeholders. A total of 67 percent of all respondents reported that benchmarking graduate health care administration programs was important. The study results revealed a significant difference between populations on the importance of evaluating certain process and outcome measures related to curriculum, research, student characteristics, and resources. However, most of the stakeholders reported that curriculum, faculty, and graduate student performance were the key quality indicators of a program. The results of this study provide preliminary information for health care administration programs to begin to develop an educational benchmarking effort. PMID- 11936770 TI - Responding to challenges in health management education. PMID- 11936771 TI - Carnegie Mellon University's MMM program: management education for 21st-century physicians. AB - The number and types of executive and graduate-level management programs for physicians have exploded in recent years. These programs take on a variety of formats, ranging from executive seminars to master's-level degree programs. Options for physicians obtaining the master's degree tend to be either regionally based programs in traditional evening classes or nationally based programs that combine executive education formats with distance education. This paper examines a nationally based program - the Master of Medical Management (MMM) - from the perspectives of an administrator and a graduate of the program. It offers reasons for the growth of similar programs and data from students enrolled in the Carnegie Mellon University MMM program. The paper also examines educational outcomes in the form of behavioral competencies that the physicians acquired in the program. It concludes with reflections on the future of the MMM and related programs for physician executives in the 21st century. PMID- 11936772 TI - King's College's Graduate Program in Health Care Administration helps to achieve community wellness: a case study. AB - Graduate programs in health care administration can become catalysts in the community wellness initiatives that the Institute of Medicine and the Centers for Disease Control and Prevention have identified as being critical to the improvement of public health among the U.S population. This paper examines the results of such a program at King's College, Wilkes-Barre, Pennsylvania, which developed community wellness programs through the establishment of a center for health promotion, assisted in the establishment of a city health department, developed a peer leader tobacco education program for elementary school students, and produced a 30-minute video on adolescent high-risk behavior. The program's goal is to facilitate coalition building among health agencies and to produce graduates equipped with administrative skills and a thorough knowledge of the value of well-developed community wellness programs. Healthy communities will require the emergence of leaders who can gather information about high-risk health behaviors and work with communities to implement solutions. Health care administration programs have a tremendous opportunity to become catalysts in the development and implementation of educational programs that may improve a community's overall health and reduce health care costs. PMID- 11936773 TI - Discrimination between cystic fibrosis and CFTR-corrected epithelial cells by a membrane potential-sensitive probe. AB - Methods to detect functional cystic fibrosis transmembrane conductance regulator (CFTR) are needed for the assessment of new therapies in cystic fibrosis (CF). We have combined patch-clamp and fluorimetric techniques to investigate whether the fluorescent voltage-sensitive probe bis-(1,3-diethylthiobarbituic anid) trimethine oxonol (Di5BAC2(3)) discriminates between changes of membrane potential (Vm evoked by cAMP in CF and CYFT-corrected epithelial cells. About 60% of the (FTR-correrced cells increased their membrane conductance and depolarized in response to cAMP, as compared to about 20% of CF cells. CFTR was found to contribute only to a fraction of the cAMP-induced responses, as indicated by the differential effects of Cl- channel blockers. Simultaneous reocording of fluorescence (AF) and membrane potential revealed that AF detected Vm changes as small as 10 mV . The relationship between deltaF and deltaVm however, was not proportional. When a large number of cells were analyzed by digital imaging, an increase in deltaF in response to cAMP was detected in the majority of CFTR corrected cells, but only in a small proportion of CF cells. The results indicate that the DiSBAC2(3) approach is a valid tool to compare cell populations with different proportions of cells responding to CFTR arcivation by cAMP. It cannot be used, however, for quantitative assessment of functional CFTR in individual CF cells. PMID- 11936774 TI - Effects of diesel exhaust on murine alveolar macrophages and a macrophage cell line. AB - To investigate the effects of diesel exhaust particles (DEP) and mycobacterial infection on macrophages, we examined protein and mRNA expression levels of various cytokines, including tumor necrosis factor-alpha (TNF-alpha), interleukin (IL)-1beta, IL-12, and IL-18 in BALB/c mouse alveolar macrophages (AM) and a macrophage cell line (RAW264.7) after in vitro stimulation with diesel exhaust (DE), with and without Mycobacterium bovis bacillus Calmette-Guerin (BCG). The cells were exposed to DE freshly generated in an in vitro system. When AM were exposed to DE (mean DEP exposure level, 300 microg/m3), the levels of TNF-alpha and IL-12 decreased significantly (by 51% to 61% for TNF-alpha and by 69% to 78% for IL-12), whereas those of IL-1beta and IL-18 remained unchanged. When AM were exposed to DE and then treated with BCG, the level of TNF-alpha decreased by 45 % to 71%, whereas the level of IL-1beta increased by 154%, compared with AM treated with BCG alone. Similarly, RAW2264.7 cells were stimulated with DE with and without M. bovis BCG and cytokine mRNA levels examined by reverse transcriptase polymerase chain reaction (R7-PCR). Longer exposure to DE decreased TNF-alpha and IL-12 mRNA levels in the RAW264.7 cells. When the cells were exposed to DE and subsequently treated with BCG, levels of TNF-alpha, IL-1beta, and IL12 mRNAs decreased compared with those of cells treated with BCG alone. These results show that DE exposure has complex and diverse effects on cytokine production by AM, and that longer exposure (>8 hours) may suppress cytokine production by AM in vitro. Longer exposure of DE may therefore suppress the host defense in the lung and may increase susceptibility to lung infections such as mycobacterial infection. PMID- 11936775 TI - Effect of antisense oligonucleotides to nuclear factor-kappaB on the survival of LPS-induced ARDS in mouse. AB - Because nuclear factor (NF)-kappaB-regulated cytokines, including tumor necrosis factor-alpha (TNF-alpha), from monocytes and macrophages have been implicated in the pathogenesis and development of septic shock and acute respiratory distress syndrome (ARDS), the effect of the antisense oligonucleotide to the p65 subunit of NF-kappaB on the survival of lipopolysaccharide (LPS)-induced ARDS in BALB/c mice was examined. None and 70% of the animals died of diffuse hemorrhagic lung edema 1 to 2.5 days after intraperitoneal administration of 10 and 20 mg/kg LPS alone, respectively. Intravenously administered antisense oligonucleotide alone did not produce any significant changes in the behavior or lung histology. After intravenous administration of the anti-sense oligonucleotide, both peripheral blood monocytes and alveolar macrophages in bronchoalveolar lavage fluid were confirmed to contain sufficiently large amounts of intracellular antisense oligonucleotides for their function usingfluorescein isothiocyanate (FTCC) labeled microscopy. The antisense oligonucleotide administered 6 hours before the intraperitoneal administration of LPS significantly decreased the survival rate with the progress of hemorrhagic edema in lung histology; 90% and 100% of animals treated with the antisense oligonuleotide died 0.5 to 1.5 days after the administration of 10 and 20 mg/kg LPS, respectively. These findings suggest that the suppression of cytokines and mediators in monocytes and alveolar macrophages by the antisense oligonucleotide to the p65 subunit of NF-kappaB worsens the survival of LPS-induced ARDS in mice with the progress of hemorrhagic lung edema. PMID- 11936776 TI - Differential expression of transforming growth factor-beta type I and II receptors by pulmonary cells in bleomycin-induced lung injury: correlation with repair and fibrosis. AB - In a rat model of lung injury induced by the antineoplastic antibiotic, bleomycin, there is loss of type I alveolar epithelial cells (AECs) followed by infiltration of activated inflammatory cells, type II AEC proliferation, and fibrosis. At 4 and 7 days after bleomycin administration alveolar macrophages have increased production and release of active transforming growth factor (TGF) beta1, an inhibitor of epithelial cell proliferation. Paradoxically at these same time intervals there is a concomitant induction of type II AEC proliferation. For TGF-beta-mediated signal transduction to occur, the expression of both TCF-beta receptor types I (TbetaR-I) and II (TbetaR-II) must be present. Using immunohistochemistry and in situ hybridization, 4 and 7 days after bleomycin administration the expression of TbetaR-I on AECs was reduced whereas that of TbetaR-II was unaltered. However, 14 and 28 days after bleomycin injury, when there is decreased proliferation and induction of differentiation of type II AECs, there was a return of TbetaR-I expression on AECs. In contrast, TbetaR-I and TbetaR-II were observed on interstitial fibroblasts at all time intervals after bleomycin administration. Because both TbetaR-I and TbetaR-II are required for signal transduction, the reduction of TbetaR-I levels on the alveolar epithelium may alter the sensitivity of AECs to the antiproliferative effects of TGF-beta1 present in increased quantities following bleomycin injury. The loss of an antiproliferative response to TGF-beta1 may be important for the regeneration of the alveolar epithelium by proliferation while the expression of both receptors onfibroblasts would result in TGF-1 signaling for the synthesis of connective tissue proteins. Ourfindings suggest that during bleomycin-induced pulmonary fibrosis, the effects of TGF-beta1 on cells may be regulated by the expression of TbetaRs. PMID- 11936777 TI - Nmda alters the development of hypoxic pulmonary vasoconstriction and nitric oxide synthetase activity in the isolated perfused rat lung. AB - Using an isolated salt-perfused rat lung model, the authors investigated whether N-methyl-D-aspartate (NMDA) (1 mM) in the pulmonary circulation effects the pulmonary vascular responses to an acute stimulus of hypoxic insult under baseline, nitric oxide synthetase (NOS)-blocked conditions (N-omega-nitro-L arginine methyl ester; L-NAME, 2 mM), and with an NMDA receptor blocker, MK-801 (0.3 microM) added. NOS activity at baseline, and in response to hypoxia, NMDA, L NAME, and a combination of these stimuli were also assessed. NMDA did not in itself alter hypoxic pulmonary vasoconstriction (HPV), but did significantly attenuate HPV during VOS blockade. This effect of NMDA was erased by MK-801. Assessment of NOS activity showed that hypoxia alone caused a doubling of NO production within the lung. This effect was erased by the addition of L-NAME. NMDA alone caused a significant, 3-fold increase in NOS activity, which was not further affected by hypoxic chalenge. L-NAME did not depress NOS activity in the hypoxia + NMDA group. These data suggest that NMDA receptor activation results in increased NOS activity and presumably increased production of NO. The increased NOS activity induced by NMDA receptor stimulation is resistant to the blockade effect of L-NAME. The actions of NMDA receptor activation may represent a natural protective mechanism, at least within the pulmonary vasculature, in face of acute, abnormal stimuli such as hypoxia. PMID- 11936779 TI - Evaluation of the focal distance of lenses by white-light Lau phase interferometry. AB - We used a white-light Lau phase interferometer to evaluate the focal distances of two lenses. We found that the variation in the experimentally measured value is less than +/-1% from the given values. Limitations of the method and error analysis are presented. PMID- 11936778 TI - Influence of dielectric substrate on the responsivity of microstrip dipole antenna-coupled infrared microbolometers. AB - We report on the influence of the dielectric substrate on the performance of microstrip dipole-antenna-coupled microbolometers. The location, the width, and the magnitude of the resonance of a printed dipole are altered when the dielectric substrate is backed by a ground plane. A thicker dielectric substrate shifts the antenna resonance toward shorter dipole lengths and leads to a stronger and slower detector response. The incorporation of an air layer into the antenna substrate further increases thermal impedance, leading to an even stronger response and shifting the antenna resonance toward longer dipole lengths. PMID- 11936780 TI - Skylab 3600 groove/mm replica grating with a scandium-silicon multilayer coating and high normal-incidence efficiency at 38-nm wavelength. AB - A Sc-Si multilayer coating was applied to a replica of the 3600 groove/mm grating, developed for the SO82A spectroheliograph that flew on the Skylab mission, for the purpose of enhancing the normal-incidence efficiency in the extreme-ultraviolet region. The efficiency, measured at an angle of incidence of 6 degrees with synchrotron radiation, had a maximum value of 7.2% at a wavelength of 38 nm and was a factor of 3 higher than the efficiency of the gold-coated Skylab grating. The measured efficiency of the Sc-Si grating was in good agreement with the efficiency calculated by use of the modified integral method. PMID- 11936781 TI - Resolution improvement in two-photon fluorescence microscopy with a single-mode fiber. AB - The dependence of spectral broadening of an ultrashort-pulsed laser beam on the fiber length and the illumination power is experimentally characterized in order to deliver the laser for two-photon fluorescence microscopy. It is found that not only the spectral width but also the spectral blue shift increases with the fiber length and illumination power, owing to the nonlinear response in the fiber. For an illumination power of 400 mW in a 3-m-long single-mode fiber, the spectral blue shift is as large as 15 nm. Such a spectral blue shift enhances the contribution from the short-wavelength components within the pulsed beam and leads to an improvement in resolution under two-photon excitation, whereas the efficiency of two-photon excitation is slightly reduced because of the temporal broadening of the pulsed beam. The experimental measurement of the axial response to a two-photon fluorescence polymer block confirms this feature. PMID- 11936782 TI - Fabrication of a graded-index polymer optical fiber preform without a cavity by inclusion of an additional monomer under a centrifugal force field. AB - When two or more monomers with different densities and refractive indices are polymerized under a centrifugal force field, a radially varying refractive index is generated owing to the difference in density of the monomers. After the polymerization is completed, a cavity is generated about the rotational axis as a result of inherent volume shrinkage during bulk radical polymerization. Therefore it is necessary to feed an additional monomer into the cavity to compensate for the undesirable volume shrinkage. We have successfully fabricated a preform with graded indices for polymer optical fiber without a cavity by adding another monomer during rotation of the reactor. One can control the overall refractive index profile by changing the rotation speed. Furthermore, the refractive-index profile can be predicted as a function of rotating speed by use of a simple mathematical model. PMID- 11936783 TI - Measurement of the induced refractive index in a photothermorefractive glass by a liquid-cell shearing interferometer. AB - A liquid-cell shearing interferometer was developed to measure refractive-index variations (delta n) in transparent materials. The cell was filled with a liquid having a matched refractive index. The achieved resolution was better than 1/1000 of a fringe shift and resulted in a delta n measurement sensitivity down to 10( 7) for 1-mm-thick samples. A refractive-index increment in photothermorefractive glass of up to 5 x 10(-6) was observed after UV exposure at 325 nm. A refractive index decrement of up to 1 x 10(-3) was observed after thermal development of the exposed sample. It was proved that photothermorefractive glass obeys the reciprocity law; i.e., delta n depends on the UV dosage but does not depend on the irradiance. PMID- 11936784 TI - Partitioned-field uniaxial holographic lenses. AB - The efficiency and aberration of partitioned-field uniaxial volume holographic compound lenses are theoretically and experimentally studied. These systems increase the image fields of holographic volume lenses, limited by the angular selectivity that is typical of these elements. At the same time, working with uniaxial systems has led to a decrease in aberration because two recording points (that behave as aberration-free points) are used. The extension of the image field is experimentally proved. PMID- 11936785 TI - Full-color imaging through a turbid medium by use of photorefractive coherence gating and a technique to separate the recording spaces. AB - We demonstrate full-color imaging through a turbid medium by use of photorefractive coherence gating and a technique to separate the recording space of each color from those of the other colors. We found that the recording spaces must be separate when a multicolor image is recorded in a photorefractive crystal to prevent the interference of the holograms with one another. For full-color imaging we used a He-Cd white-light laser, which is compact and useful for full color holography. Full-color-image retrieval is demonstrated through five mean free paths of a turbid medium. PMID- 11936786 TI - Low-loss ultrabroadband 90 degree optical rotator with collinear input and output beams. AB - An achromatic device to rotate optical polarization by 90 degrees is described. This is based on a series of reflecting surfaces that rotates incoming light about the optical axis and translates it such that the exiting light is collinear. Polarization rotation is achieved by rotation of the optical beam, as opposed to the more common approach of phase retardation by use of birefringent elements. For broadband operation from the UV to the near infrared, the device was constructed by use of total internal reflection in three fused-silica glass components. Losses are minimized with interstitial surfaces designed to be angled close to Brewster's angle. PMID- 11936787 TI - Ellipsometric investigation of rough zinc arsenide epilayers. AB - We report single-wavelength ellipsometric measurements of the complex index of refraction of rough Zn3As2 films on InP substrates. What we believe to be a novel technique, based on surface roughness measurements by atomic-force microscopy, is discussed to extract useful information from the ellipsometry results. The anticipated presence of a thin oxide layer is confirmed by Auger electron spectroscopy. PMID- 11936788 TI - Rainbow refractometry: simultaneous measurement of temperature, refractive index, and size of droplets. AB - The capabilities and limitations of rainbow refractometry, specifically the minimum measurable droplet size and the errors in the refractive index (temperature), have been studied. We evaluate what we believe is a new method of indirectly applying the Lorenz-Mie theory to rainbow refractometry. The results show that this new method reduces the errors and eliminates the biases that may occur if the Airy theory is used. A more precise method to filter the high frequency oscillations associated with the measurement signals was developed. Finally, it was discovered that the errors associated with rainbow refractometry are such that a single droplet measurement is unreliable. A mean refractive index should be determined on the basis of multiple droplet measurements. PMID- 11936789 TI - Fiber-optic infrared radiometer for accurate temperature measurements. AB - A fiber-optic radiometer is developed for accurate noncontact temperature measurements. Of compact and novel design, it is based on replacing the usual chopper with a simple shutter. The radiometer operates in a spectral range of 5 20 microm and uses a silver-halide IR-transmitting optical fiber. The radiometer has a temperature resolution of 0.1 degrees C, a time response of 200 ms, and a spatial resolution of approximately 1 mm. Theory, simulation, radiometer design and construction, and examples of experimental measurements are shown. The novel radiometer can be used in diverse applications in science, medicine, and industry. PMID- 11936790 TI - Interferometric phase reconstruction by nonuniform shifting of the reference beam. AB - A method for phase measurement in common-path interferometers, believed to be novel, is presented. We use the property of phase reconstruction algorithms, such as the Carre and Hariharan algorithms, that do not require uniform phase across the reference beam. Only the ratio of the phase steps must be the same at each pixel. We show phase measurement and reconstruction in a common-path interferometer by shifting either the tilt or the focus of the reference wave front. We present a theoretical explanation of phase measurement using this property. We also present results from a proof-of-principle experiment using a scatterplate interferometer, in conjunction with the tilt phase-shifting technique, to measure the reflected phase of a test optical element. Furthermore, we present a computer simulation to demonstrate the mathematical validity of this measurement technique using defocus shifting, rather than tilt shifting, in the reference wave front. PMID- 11936791 TI - Temperature measurements in steady axisymmetric partially premixed flames by use of rainbow schlieren deflectometry. AB - We focus on the utility of rainbow schlieren as a tool for measuring the temperature of axisymmetric partially premixed flames (PPFs). Methane-air PPFs are established on a coannular burner. The flames involve two spatially distinct reaction zones, one in an inner premixed region that has a curved tip and a spatially planar wing portion and another that involves an outer nonpremixed zone in which intermediate species burn in air. Schlieren images are found to visualize clearly these PPF characteristics through light deflection by steep refractive-index gradients in the two reaction zone fronts. The temperature distributions of two flames established at fuel-rich mixture equivalence ratios of phi(r) = 1.5 and 2.0, with bulk-averaged velocities, Vreac = 60 cm s(-1) and Vair = 50 cm s(-1), are inferred from color schlieren images, and a measurement error analysis is performed. Errors arise from two sources. One lies in the process of inferring the temperature from the refractive-index measurement by making assumptions regarding the local composition of the flame. We have shown through simulations that the average temperature deviations due to these assumptions are 1.7% for the phi(r) = 1.5 flame and 2.3% for the phi(r) = 2.0 flame. Another source involves the local uncertainty in the measurement of the transverse ray displacement at the filter plane that is used to determine the refractive index and thereafter the flame temperature. We have ascertained that a maximum error of 4.3% in the temperature determination can be attributed to this local measurement uncertainty. This investigation demonstrates the capability of the schlieren technique for providing not only qualitative displays of the PPFs but also full-field-of-view temperature measurements that are accurate, spatially resolved, and nonintrusive. PMID- 11936792 TI - Study of bandwidth effects in monochromator-based spectral responsivity measurements. AB - The errors caused by monochromator bandwidth in spectral responsivity measurements with a monochromator-based apparatus are discussed. Bandwidth effects are not negligible in high-accuracy cryogenic radiometer-based calibrations. A simple numerical method is used to calculate bandwidth effects for different types of detectors, monochromator slit scattering functions, and monochromator output spectral distributions. The method uses low-order Lagrange polynomials fitted segmentwise to measured spectral responsivity and monochromator spectral distribution data in order to make the calculations. It is shown that the shape of the slit function has only a small influence on the bandwidth errors, whereas the output spectral distribution of the monochromator can strongly affect bandwidth errors. It is also shown that in most cases the magnitude of bandwidth effects will vary as the square of the bandwidth. Bandwidth error calculations are presented for various types of detectors (silicon, silicon trap, germanium, InGaAs), for a V(X) detector, and for a typical filter radiometer. A comparison is made between calculated and measured bandwidth effects to validate the method used. In general, calculations of bandwidth effects will be mostly useful for determining uncertainties associated with monochromator bandwidth in spectral responsivity measurements; however, in certain cases the calculations can be used to apply corrections for such effects. PMID- 11936793 TI - Design of angle-tolerant multivariate optical elements for chemical imaging. AB - Multivariate optical elements (MOEs) are multilayer optical interference coatings with arbitrary spectral profiles that are used in multivariate pattern recognition to perform the task of projecting magnitudes of special basis functions (regression vectors) out of optical spectra. Because MOEs depend on optical interference effects, their performance is sensitive to the angle of incidence of incident light. This angle dependence complicates their use in imaging applications. We report a method for the design of angle-insensitive MOEs based on modification of a previously described nonlinear optimization algorithm. This algorithm operates when the effects of deviant angles of incidence are simulated prior to optimization, which treats the angular deviation as an interferent in the measurement. To demonstrate the algorithm, a 13-layer imaging MOE (IMOE, with alternating layers of high-index Nb2O5 and low-index SiO2) for the determination of Bismarck Brown dye in mixtures of Bismarck Brown and Crystal Violet, was designed and its performance simulated. For angles of incidence that range from 42 degrees to 48 degrees, the IMOE has an average standard error of prediction (SEP) of 0.55 microM for Bismarck Brown. This compares with a SEP of 2.8 microM for a MOE designed by a fixed-angle algorithm. PMID- 11936794 TI - Near-field effects in multilayer inductive metal meshes. AB - The transmittance of inductive single-layer and multilayer cross-shaped metal meshes has been calculated with the Micro-Stripes software program. The effect of symmetric and asymmetric alignment of the crosses of one mesh with respect to another was studied and compared with transmission line theory, which presents the nonaligned case. Significant differences are found for small spacing at approximately 1/5 the periodicity constant, whereas the differences disappear for large spacing. A pair of coupled surface waves is used to represent the mode of a single mesh. The resulting modes corresponding to the transmittance of multilayer metal meshes are interpreted by modes composed of resonance modes of a single mesh coupled by Fabry-Perot modes depending on the separation. PMID- 11936795 TI - Disturbance-free high-speed sinusoidal phase-modulating laser diode interferometer. AB - A surface profiler that incorporates a feedback controller to eliminate external disturbances is proposed and demonstrated. Its overall performance is dependent on the frequency response of the feedback loop. The frequency of the modulating signal strongly influences the response of the feedback controller. When we used the integrating-bucket method, the CCD camera had to be operated at a low frequency video rate. Our technique uses a CCD camera equipped with an electronic shutter. The shutter function enables us to apply high-speed sinusoidal phase modulation to the conventional integrating-bucket method under the standard video rate. PMID- 11936796 TI - Improved Saunders method for the analysis of lateral shearing interferograms. AB - An interferogram obtained by use of ordinary interferometers, such as Fizeau and Twyman-Green interferometers, will show a contour map of the wave front under test. A lateral-shearing interferogram, however, will show a contour map of the difference between the wave front under test and a sheared wave front, that is, a contour map of the derivative of the wave front under test. Therefore one can reconstruct the shape of the wave front under test by analyzing that difference. Many methods for reconstructing a wave front have been proposed. The Saunders method reconstructs a wave front; rapidly however the wave-front data are reconstructed only at intervals of the amount of shear along the direction of the shear. Therefore the method has low spatial resolution. A method for reconstructing a wave front that is based on the Saunders method and has high spatial resolution is proposed. The method analyzes the differences that are produced by shearing of the wave front under test in many directions. This method requires a large number of interferograms for reconstructing the wave front. Here the method is described, and its validity is confirmed by simulation. PMID- 11936797 TI - Synthesis of longitudinal coherence functions by spatial modulation of an extended light source: a new interpretation and experimental verifications. AB - Giving a new physical interpretation to the principle of longitudinal coherence control, we propose an improved method for synthesizing a spatial coherence function along the longitudinal axis of light propagation. By controlling the irradiance of an extended quasi-monochromatic spatially incoherent source with a spatial light modulator, we generated a special optical field that exhibits high coherence selectively for a specific pair of points at specified locations along the axis of beam propagation. This function of longitudinal coherence control provides new possibilities for dispersion-free measurements in optical tomography and profilometry. A quantitative experimental proof of principle is presented. PMID- 11936798 TI - Two-wavelength laser diode interferometer with time-sharing sinusoidal phase modulation. AB - We describe an interferometer system that uses two separate wavelengths to measure step height. The overlapping interference images detected by a CCD camera are easily separated by an ordinary integrating-bucket method and time-sharing sinusoidal phase modulation, in which two laser diodes are alternately modulated with a sinusoidal signal. A phase map is obtained only for the laser diode into which the modulation signal is injected. In this instance, a 1-microm step height was accurately detected. PMID- 11936799 TI - Application of fluorescence microscopy for noninvasive detection of surface contamination and precursors to laser-induced damage. AB - We present an experimental investigation to evaluate fluorescence microscopy as a tool to detect surface contamination as well as reveal surface damage precursors on optical components for large-aperture laser systems. We performed fluorescence imaging experiments using 351-nm laser excitation, whereas in situ damage testing was performed at laser fluences well below the dielectric breakdown threshold of the pure material. The experimental results demonstrated the potential of this technique to address both aforementioned technical issues. PMID- 11936800 TI - Complex methylene-blue-sensitized polyvinyl chloride: a polymer matrix for hologram recording. AB - A new polymer matrix sensitized with methylene blue for use as an optical recording material is described here. The characterization is done to determine the optimal recording conditions. These films need no chemical development and are found to be stable for several months. The matrix has excellent shelf life and needs an exposure only as short as 20 s. Direct imaging was done on this material. PMID- 11936801 TI - Elasto-optics in double-coated optical fibers induced by axial strain and hydrostatic pressure. AB - Stresses, microbending loss, and refractive-index changes induced simultaneously by axial strain and hydrostatic pressure in double-coated optical fibers are analyzed. The lateral pressure and normal stresses in the optical fiber, primary coating, and secondary coating are derived. Also presented are the microbending loss and refractive-index changes in the glass fiber. The normal stresses are affected by axial strain, hydrostatic pressure, material properties, and thickness of the primary and secondary coatings. It is found that microbending loss decreases with increasing thickness, the Young's modulus, and the Poisson's ratio of the secondary coating but increases with the increasing Young's modulus and Poisson's ratio of the primary coating. Similarly, changes in refractive index in the glass fiber decrease with the increasing Young's modulus and Poisson's ratio of the secondary coating but increase with the increasing Young's modulus and Poisson's ratio of the primary coating. Therefore, to minimize microbending loss induced simultaneously by axial strain and hydrostatic pressure in the glass fiber, the polymeric coatings should be suitably selected. An optimal design procedure is also indicated. PMID- 11936802 TI - Third-harmonic generation microscopy in highly scattering media: comment. AB - In their contribution "Third-harmonic generation microscopy in highly scattering media" [Appl. Opt. 39, 5187 (2000)], C. M. Blanca and C. Saloma apply the Monte Carlo technique to the calculation of third-harmonic generation (THG) in strongly scattering samples. In that study the authors fail to appreciate the coherent nature of both the signal-generation and the image-formation process, inherent to THG microscopy. Here we wish to clarify several points. PMID- 11936803 TI - Thermal properties and crystallization dynamics of a phase-change alloy for write once optical data storage. AB - Using a two-laser static tester, we measured the crystallization temperature and the thermal conductivity of a phase-change alloy thin film used in write-once read-many media of optical data storage. The experimental technique, in general, and the calibration procedures, in particular, are described. The measurement results are used as entry points into numerical calculations that ultimately yield estimates of the material parameters. Valuable information about the dynamics of mark formation (i.e., localized crystallization) in amorphous phase change alloy films is obtained from the observed variations of the sample reflectance under short-pulse and long-pulse recording conditions. The dependence of these reflectance variations on the laser pulse power has also been investigated. PMID- 11936804 TI - Aspect ratio of elongated polychromatic far-field speckles of continuous and discrete spectral distribution with respect to surface roughness characterization. AB - Here polychromatic speckle patterns generated either by a polychromatic light source that emits at discrete frequencies or by a light source showing a continuous narrow-band spectral distribution are studied. The purpose here is the application of polychromatic speckle-pattern analysis to an in-process surface roughness characterization. To compare the coherence properties of the different polychromatic light sources, first a modified definition of the coherence length is introduced. Furthermore, the relevant optical phenomena, namely, the speckle elongation caused by the angular dispersion and the roughness-dependent speckle decorrelation, are summarized. It is shown that light sources with a continuous spectral distribution have essential advantages in comparison with discrete wavelength sources. The theoretical results are confirmed by experimental investigations based on a digital algorithm for the evaluation of CCD images of polychromatic speckle patterns, which are recorded in the Fourier plane of a Fourier-transforming lens. PMID- 11936805 TI - Compact method for optical induction of proximal probe heating and elongation. AB - A tapered, metal-coated, optical fiber probe will elongate when heated by light input through a fiber. The induced motion can be used for data storage or nanostructuring of a surface. The elongation produced by this alignment-free system is measured with force feedback in a near-field scanning optical microscope (NSOM). The input light intensity controls the elongation magnitude, which ranges from a few nanometers to more than 100 nm. A 0.5-mW input energy yields approximately 20 nm of probe elongation. The elongation quantified here can create artifacts in any experiment using pulsed laser light with a NSOM or an atomic force microscope. PMID- 11936806 TI - Speckle size of light scattered from slightly rough cylindrical surfaces. AB - This research is an extension of the optical method of quality control presented in a previous paper [Appl. Opt. 39, 5811 (2000)] to the case of slightly rough cylindrical surfaces. Applying the Kirchhoff scalar diffraction theory yields an analytical expression of the autocorrelation function of the intensity scattered from slightly rough cylindrical surfaces. This function, which is related to speckle size and shape, is shown to depend on the surface correlation length, unlike for plane surfaces for which the speckle depends on the illuminated area only. The theoretical expression is compared with that for the speckle produced by the light scattered from a cylindrical bearing and from various high-quality wires, showing that the method allows the correlation lengths of high-quality cylindrical surfaces to be determined. PMID- 11936807 TI - Characterization of milk properties with a radiative transfer model. AB - To characterize milk through light-scattering measurements, a semianalytical radiative transfer model was used to simulate the backscatter of light in milk having homogenized fat levels from 0.05 to 3.2 wt. %. The input parameters to the model include the incident wavelength, refractive index of particles and medium, and particle number densities. By varying the wavelength, we can obtain a reasonable fit between experimental data and the model for lower fat milks. Results indicate that the model is most sensitive to the particle diameter and size distribution and less sensitive to the number and index of refraction of the particles. PMID- 11936808 TI - Refractive-index profiling of embedded microstructures in optical materials. AB - We describe use of a phase-sensitive low-coherence reflectometer to measure spatial variation of refractive index in optical materials. The described interferometric technique is demonstrated to be a valuable tool to profile the refractive index of optical elements such as integrated waveguides and photowritten optical microstructures. As an example, a refractive-index profile is mapped of a microstructure written in a microscope glass slide with an ultrashort-pulse laser. PMID- 11936809 TI - Internal stress and optical properties of Nb2O5 thin films deposited by ion-beam sputtering. AB - The influence on the internal stress and optical properties of Nb2O5 thin films with ion-beam energy was investigated. Nb2O5 thin films were deposited on unheated glass substrates by means of ion-beam sputtering with different ion-beam voltage, Vb. The refractive index, extinction coefficient, and surface roughness were found to depend on the ion-beam energy. The stresses in thin films were measured by the phase-shifting interferometry technique. The film stress was also found to be related to Vb, and a high compressive stress of -0.467 GPa was measured at Vb = 850 V. The Nb2O5-SiO2 multilayer coatings had smaller average compressive stress as compared with single-layer Nb2O5 film. PMID- 11936810 TI - Chromium-scandium multilayer mirrors for the nitrogen K(alpha) line in the water window region. AB - Chromium-scandium (Cr-Sc) is a promising material combination for multilayer mirrors in the water window region. A possible x-ray source for laboratory use in this wavelength range is the nitrogen K(alpha) line at 3.16 nm. High reflectivities at this wavelength can be achieved with Cr-Sc multilayer mirrors if the interfaces between adjacent layers are smooth. The growth parameters of the magnetron sputtering process for these materials have been optimized. It is shown that the reflectivity of such mirrors can be considerably improved by the application of a proper bias voltage during film growth. The high quality of the multilayer films is demonstrated with copper K(alpha) x-ray reflection and transmission electron microscopy. The reflective properties of the multilayers close to the nitrogen K(alpha) line were measured with synchrotron radiation for different angles of incidence. Reflectivities between R = 5.9% for near-normal incidence (theta = 1.5 degrees) and R = 29.6% for theta = 59.9 degrees were measured. PMID- 11936811 TI - Optical design of a solar flux homogenizer for concentrator photovoltaics. AB - An optical solution is described for the redistribution of the light reflected from a 400-m2 paraboloidal solar concentrating dish as uniformly as possible over an approximately 1-m2 plane. Concentrator photovoltaic cells will be mounted at this plane, and they require a uniform light distribution for high efficiency. It is proposed that the solar cells will be mounted at the output of a rectangular receiver box with reflective sidewalls (i.e., a kaleidoscope), which will redistribute the light. I discuss the receiver box properties that influence the light distribution reaching the solar cells. PMID- 11936812 TI - Wide-field optical coherence tomography: imaging of biological tissues. AB - We describe a two-dimensional optical coherence tomography technique with which we were able to obtain multiple longitudinal slices of a biological sample directly in a single Z scan. The system is based on a femtosecond Cr4+:forsterite laser and an infrared camera for wide-field imaging of the sample with a depth resolution of 5 microm. With this imaging apparatus we were able to investigate human skin and mouse ear samples and to observe the different constitutive tissues. PMID- 11936813 TI - Accuracy analysis in laser keratopography. AB - We analyze the accuracy of a laser keratopographer in the evaluation of corneal topography for non-smooth corneal surfaces and when some noise is introduced into measured data. Through some numerical simulations, cosinesoidal deformations with different amplitudes and spatial periods are introduced on theoretical surfaces. Gaussian noise is introduced on the simulated x and y position coordinates for the measured position of the reflected beam in order to simulate detection errors that are due to vibrations or electric and other noise on the position-sensing detector. We found that the topography of the surface could be obtained with reliable accuracy if the height-to-width ratio of the deformations of the surface is smaller than 0.02 and the error in the detection of position at the position sensing detector is under 0.5 mm. PMID- 11936814 TI - Integrated planar terahertz resonators for femtomolar sensitivity label-free detection of DNA hybridization. AB - A promising label-free approach for the analysis of genetic material by means of detecting the hybridization of polynucleotides with electromagnetic waves at terahertz (THz) frequencies is presented. Using an integrated waveguide approach, incorporating resonant THz structures as sample carriers and transducers for the analysis of the DNA molecules, we achieve a sensitivity down to femtomolar levels. The approach is demonstrated with time-domain ultrafast techniques based on femtosecond laser pulses for generating and electro-optically detecting broadband THz signals, although the principle can certainly be transferred to other THz technologies. PMID- 11936815 TI - Methods for parallel-detection-based ultrasound-modulated optical tomography. AB - The research reported here focuses on ultrasound-modulated optical tomography based on parallel speckle detection. Four methods were investigated for signal acquisition and analysis, in which laser speckle statistics was applied. The methods were compared with the previously used four-phase method in the imaging of all-biological-tissue samples, in which the buried objects were also biological tissues. The image quality obtained with these methods was comparable with that obtained with the four-phase method; in addition, these methods have advantages in reducing acquisition time and improving the signal-to-noise ratio. PMID- 11936816 TI - Projecting SEER cancer survival rates to the US: an ecological regression approach. AB - OBJECTIVES: Cancer survival information is available only in areas covered by cancer registration. The objective of this study is to project cancer survival for the entire US as well as states from survival data from the National Cancer Institute's Surveillance, Epidemiology, and End Results (SEER) program. METHODS: Five-year breast, prostate, and colorectal cancer relative survival rates from SEER are regressed on socioeconomic, demographic, and health variables at the county level. These models are first validated by comparing the observed rates with projected rates for counties not used in the estimation process. RESULTS: Education was the best indicator of longer cancer survival. Other important predictors of the geographical variability of survival varied by cancer site. Better survival was predicted for breast and prostate than for colorectal cancer. CONCLUSIONS: Data from cancer registries can be used in ecological models to provide national and state estimates of patients' survival rates. These estimates are useful in targeting areas in which to promote earlier diagnosis or improved access to care, and may also aid in monitoring the quality of survival data collected by individual cancer registries. PMID- 11936817 TI - Galactose-1-phosphate uridyl transferase (GALT) genotype and phenotype, galactose consumption, and the risk of borderline and invasive ovarian cancer (United States). AB - OBJECTIVE: Previous studies have suggested that high levels of galactose consumption and/or low levels of galactose-I-phosphate uridyl transferase (GALT) activity may result in an increased risk of epithelial ovarian cancer. Similarly, some have reported that carriers of the N314D (asparagine at codon 314 replaced by aspartate) GALT polymorphism, which can be associated with low GALT activity, may have a higher risk of ovarian cancer. We examined these issues as part of a large case-control study of ovarian cancer conducted in Los Angeles between 1992 and 1998. METHODS: A total of 1,439 histologically confirmed borderline and invasive ovarian cancer cases among English-speaking non-Asian women were ascertained through the population-based cancer registry for Los Angeles County and completed in-person interviews were obtained from 689 of these (78% of cases approached). Controls consisted of 645 English-speaking non-Asian women with at least one intact ovary matched to cases on race/ethnicity (African-American, Latina, non-Latina White), date of birth (+/-3 years), and neighborhood of residence. Interviewer-administered questionnaires included information on reproductive factors, exogenous hormone use, medical history, and diet. Dietary information for the year before each case's diagnosis (and the same period for her matched control) was obtained using a self-administered food-frequency questionnaire. Blood samples were obtained from 452 controls, 136 cases with borderline ovarian cancer, and 312 cases with invasive ovarian cancer. The N314D polymorphism was characterized using PCR-RFLP and GALT enzyme activity, and was determined for a sample of the subjects with GALT genotype using an erythrocyte based radioactive enzyme assay. RESULTS: We found no effect of N314D GALT genotype on the risk of borderline ovarian cancer (odds ratio (OR)=0.91; 95% confidence interval (CI)=0.54-1.6) or invasive ovarian cancer (OR=0.78; 95% CI= 0.53-1.2). Neither did we observe a relationship between GALT activity or lactose/galactose intake and risk of borderline or invasive ovarian cancer. Among N314D carriers, galactose consumption was associated with an increased risk of borderline (OR = 2.7, p = 0.01), but not invasive (OR = 1.2, p = 0.34), ovarian cancer; however, this result was based on only 24 N314D-positive borderline cases. CONCLUSIONS: Differences in galactose intake and GALT metabolism do not contribute significantly to the risk of ovarian cancer. There is some evidence that galactose intake may play a role in the development of borderline ovarian cancer among women who carry the uncommon GALT N314D polymorphism. More data are needed if this latter suggestion is to be definitively addressed. PMID- 11936818 TI - Reappraisal of the strong association between simian virus 40 and human malignant mesothelioma of the pleura (Belgium). AB - OBJECTIVE: The frequent association of a monkey oncogenic virus, simian virus 40 (SV40), with pleural mesothelioma and the proposed transmission of the virus from contaminated polio vaccines to humans has received considerable scientific and public attention. We sought to determine whether SV40 would indeed be present in mesothelioma patients from Belgium, as claimed in former studies, and to characterize the viral genome in respective specimens. METHODS: DNA was extracted from frozen tissue from 12 patients diagnosed with pleural mesothelioma in Belgium. Five different extraction methods were compared and primer pairs targeting four informative regions of the SV40 genome were used to amplify viral products by the polymerase chain reaction (PCR). One of the pairs additionally allows amplification of human polyoma viruses. Southern blotting with an oligonucleotide probe directed at the expected SV40 sequences was also applied. RESULTS: One of the 12 samples contained amplifiable JC virus DNA. In contrast, none of the samples (0/12, 95% CI 0% to 26.5%) was positive for SV40 DNA sequences. The Southern blot analysis confirmed the absence of trace amounts of SV40 PCR product. We also clearly demonstrate that an otherwise specific probe against SV40 could easily hybridize in quite a non-specific manner under recommended conditions. CONCLUSIONS: These results provide strong evidence for a lack of association of SV40 with most pleural mesotheliomas in the Belgian population. We recommend a re-examination of other positive case series and avoidance of questionable hybridization practices in future studies. PMID- 11936819 TI - Menstrual and reproductive factors and risk of non-Hodgkin lymphoma: the Iowa women's health study (United States). AB - BACKGROUND: Endogenous sex hormones, particularly estrogens, modulate the immune system, and non-Hodgkin lymphoma (NHL) is a tumor that is related to immunologic status. METHODS: Self-reported menstrual and reproductive history and risk of NHL were evaluated in a cohort of 37,934 Iowa women who were aged 55-69 years when first enrolled in 1986. Through 1998, 261 cases of NHL were identified by linkage to the Iowa SEER Cancer Registry. RESULTS: After multivariate adjustment there was no association between NHL incidence and age at menarche, age at menopause, type of menopause, history of infertility, number of miscarriages, or history of induced abortion, while there were suggestive inverse associations with nulliparity (RR=0.65; 95% CI 0.36-1.16) and years of ovulation (RR = 0.76 for >37 compared to <28 ovulatory years: p-trend = 0.07). Among parous women there was no association with number of livebirths or age at first livebirth, but there was an inverse association with number of children who were breast-fed (RR=0.52 for breast-feeding >2 children versus none; 95% CI 0.33-0.82). CONCLUSIONS: Overall, menstrual and reproductive factors were not strongly related to NHL incidence. The inverse association with breast-feeding is novel but requires confirmation in other studies. PMID- 11936820 TI - Breast and cervical cancer screening and disease incidence and stage in New Mexico. AB - OBJECTIVE: Data on the health impact of breast and cervical cancer screening programs for low-income women are limited. We sought to determine whether a statewide program to provide breast and cervical cancer screening services influenced trends in disease incidence and stage. METHODS: We assessed trend data regarding breast and cervical cancer incidence and stage gathered by the New Mexico Tumor Registry and the Surveillance, Epidemiology, and End Results Program before (1975 through 1990) and during (1991 through 1998) implementation of the New Mexico Breast and Cervical Cancer Screening Program. RESULTS: The incidence of cervical carcinoma in-situ increased rapidly in 1991. The incidence per 100,000 population for in-situ breast cancer (2.9 in 1975-1982, 8.5 in 1983-1990, and 16.8 in 1991-1998) and local breast cancer (39.9 in 1975-1982, 46.5 in 1983 1990, and 61.3 in 1991-1998) also increased during the program operation. Notably, a significant increase in incidence per 100,000 population for regional and distant breast cancer from 1975-1982 (31.5) to 1983-1990 (36.0) declined during the 1991-1998 (33.2) period of program operation. CONCLUSIONS: The statewide screening program improved detection of breast and cervical cancer and helped reduce the incidence of advanced stages of breast cancer in a relatively short time period. PMID- 11936821 TI - Sinonasal cancer and occupational exposures: a pooled analysis of 12 case-control studies. AB - OBJECTIVE: In order to examine the associations between sinonasal cancer and occupational exposures other than wood dust and leather dust, the data from 12 case-control studies conducted in seven countries were pooled and reanalyzed. METHODS: The pooled data set included 195 adenocarcinoma cases (169 men and 26 women), 432 squamous cell carcinomas (330 men and 102 women), and 3136 controls (2349 men and 787 women). Occupational exposures to formaldehyde, silica dust, textile dust, coal dust, flour dust, asbestos, and man-made vitreous fibers were assessed with a job-exposure matrix. Odds ratios (ORs) were adjusted for age, study, wood dust, and leather dust, or other occupational exposures when relevant. 95% confidence intervals (CIs) were estimated by unconditional logistic regression. RESULTS: A significantly increased risk of adenocarcinoma was associated with exposure to formaldehyde. The ORs for the highest level of exposure were 3.0 (Cl = 1.5-5.7) among men and 6.2 (CI=2.0-19.7) among women. An elevated risk of squamous cell carcinoma was observed among men (OR=2.5, CI=0.6 10.1) and women (OR = 3.5, CI = 1.2-10.5) with a high probability of exposure to formaldehyde. Exposure to textile dust was associated with non-significantly elevated risk of adenocarcinoma, among women only: the OR for the high level of cumulative exposure was 2.5 (CI = 0.7-9.0). High level of asbestos exposure was associated with a significantly increased risk of squamous cell carcinoma among men (OR = 1.6, CI = 1.1-2.3). CONCLUSIONS: The results of this pooled analysis support the hypothesis that occupational exposure to formaldehyde increases the risk of sinonasal cancer, particularly of adenocarcinoma. They also indicate an elevated risk of adenocarcinoma among women exposed to textile dust, and suggest that exposure to asbestos may increase the risk of squamous cell carcinoma. PMID- 11936823 TI - Breast cancer risk after childbirth in young women with family history (Denmark). AB - OBJECTIVE: The increased risk of breast cancer in women with family history of breast cancer (FHBC) might be reduced by early childbirths. However, childbirth induces a transient increase in risk in the first 5-10 years, which coincides with the relatively increased risk of family cases at a young age. The objective was to investigate this short-term change in risk according to FHBC. METHODS: We used a population-based cohort of 1.5 million Danish women. Between 1968 and 1990, 2770 incident cases of breast cancer below 40 years of age were identified in the Danish Cancer Register, of whom 276 (10%) had a FHBC. RESULTS: The first 5 years after a birth the short-term increase in risk was 30% (3-64%) larger in women with FHBC than without FHBC. After the first 5 years we observed no difference in the effect of a birth between women with and without FHBC. CONCLUSIONS: The adverse short-term effect of childbirthis stronger in women with FHBC. PMID- 11936822 TI - A case-control study of tobacco use and other non-occupational risk factors for t(14;18) subtypes of non-Hodgkin's lymphoma (United States). AB - OBJECTIVE: Non-Hodgkin's lymphoma (NHL) encompasses diverse subtypes, and analyzing NHL as a single outcome may mask associations. In a new approach we evaluated associations with subtypes defined by the t(14;18) translocation, reasoning that cases within these subtypes would have more common risk factors than all NHL combined. METHODS: Archival biopsies from cases in a population based NHL study were assayed for t(14;18) using polymerase chain reaction amplification. Exposures in 68 t(14;18)-positive and 114-negative cases were compared with 1245 controls. The expectation-maximization algorithm was used to fit polytomous regression models based on all available information, including data from 440 unclassified cases. RESULTS: Family history of hemolymphatic cancer was associated with t(14;18)-negative NHL (odds ratio (OR) 2.4, 95% confidence interval (CI) 1.4 3.9). but not t(14;18)-positive NHL. Cigarette smoking was weakly associated with t(14;18)-positive NHL (OR 1.7, CI 0.9-3.3), but ORs decreased as smoking increased. Chewing tobacco was associated with t(14;18) positive NHL, particularly when used before age 18 (OR 2.5. CI 1.0-6.0, 13 exposed cases). Odds ratios for both case-subtypes were doubled among hair-dye users. CONCLUSIONS: Cigarette smoking was not clearly associated with t(14;18) positive NHL. Family history may be a marker for factors that act specifically through t(14;18)-negative pathogenic mechanisms. PMID- 11936824 TI - Practical implications of imposing a new world standard population. AB - OBJECTIVE: The World Health Organization (WHO) has recently introduced a new world standard population for the production of age-standardized rates. In this study we compare cancer rates standardized to this population with those computed using reference populations in current practice, particularly the world standard of Segi (1960), in order to evaluate their adequacy as estimators of relative risk in diverse population groups and over time. METHODS: Incidence and mortality rates standardized using these reference populations were calculated and compared for various cancers. Standardized rate ratios were compared with more efficient methods of approximating relative risk, the Mantel & Haenszel and maximum likelihood estimators. The differences were tested by taking a synthesis of the relative risks and by taking into account whether effects were homogeneous across age strata or not. RESULTS: There were no statistically significant differences between the relative risk estimates based on direct standardization and those obtained using Mantel & Haenszel (p < 0.99), or maximum-likelihood techniques (p > 0.99), regardless of whether the Segi or the WHO world population was used as the standard. CONCLUSIONS: Ratios of rates age-standardized using the world standard of Segi approximate relative risk as precisely as the WHO standard. For this, and important practical reasons, it is considered unnecessary to replace the Segi standard population for comparisons between cancer rates. PMID- 11936825 TI - Paternal preconceptional irradiation, population mixing and solid tumors in the children of radiation workers (England). AB - OBJECTIVE: To investigate whether there was an increased incidence of solid tumors among offspring of male radiation workers at the Sellafield nuclear installation in Cumbria, northwest England and whether paternal preconceptional irradiation was associated with the risk of solid tumors. METHODS: A cohort study of 266,710 live births in Cumbria, 1950-1991, followed up to age 25 years on the end of 1991. RESULTS: Children of radiation workers had a non-significantly increased risk of solid tumors (RR= 1.5, 95% CI: 0.9-2.4, p = 0.09), determined largely by an increased risk of cancers excluding leukemias, lymphomas, brain, spinal and gender-specific tumors (RR= 1.9, 95% CI: 1.0-3.3, p = 0.05), which was partly explained by differing patterns of parental migration (adjusted RR= 1.7, 95% Cl: 0.8-3.2, p = 0.50). Within children of radiation workers there was no evidence of an increased risk with increasing paternal preconception dose of external radiation (hazard ratio per 100 mSv for all solid tumors=0.6, 95% CI: 0.1-1.8, p = 0.52). CONCLUSIONS: Any observed excess of solid tumors in children of radiation workers may be partly explained by population mixing. Fathers' occupational exposure to radiation before conception was not found to be risk factor for solid tumors in their children. PMID- 11936826 TI - A prospective study of life satisfaction, neuroticism and breast cancer risk (Finland). AB - OBJECTIVE: To prospectively investigate whether life satisfaction and neuroticism, two measures reflecting aspects of anxiety/depression, are related to risk of breast cancer. METHODS: The study population comprised 12,032 women from the population-based Finnish Twin Cohort. Life satisfaction and neuroticism were assessed in health questionnaires in 1975 and 1981. During 21 years of follow-up, 238 cases of breast cancer were documented in the study cohort through record linkage to the Finnish Cancer Registry. RESULTS: After adjustment for age and other important covariates, the hazard ratios were 1.0 (reference), 0.9 (95% confidence interval 0.6-1.3) and 1.0 (0.7-1.6) by decreasing level of life satisfaction (satisfied, intermediate, dissatisfied), and 1.0 (reference), 1.0 (0.7-1.4) and 0.8 (0.6-1.2) by increasing level of neuroticism (low, intermediate, high). These results did not change after accounting for possible changes in the levels of life satisfaction and neuroticism between 1975 and 1981. The null results were also confirmed with the use of a nested case-control study within twin pairs that became discordant for breast cancer during the follow-up. CONCLUSIONS: Our data indicate that life satisfaction and neuroticism are not substantially associated with breast cancer risk. These findings provide epidemiological evidence against the doubts that breast cancer would be more likely to occur in unhappy, dissatisfied, and worrying women. PMID- 11936827 TI - Postural discomfort and perceived exertion in standardized box-holding postures. AB - To help in the design or redesign of workplaces it would be helpful to know in advance the postural stress consequences of a wide range of body postures. This experiment evaluated 168 postures chosen to represent those in the Ovako Working posture Analysing System (OWAS) using Rated Perceived Exertion (RPE) and Body Part Discomfort (BPD) measures. The postures comprised all combinations of three arm postures, four back postures, seven leg postures and two forces (weights of held boxes). Twelve male subjects held each posture for a fixed duration (20 s) before providing RPE and BPD ratings. Analysis of the ratings gave highly significant main effects, with the major driver being the object weight. As each factor was varied, the largest effect was on the body region corresponding to that factor. A simple main-effects-only additive model explained 91% of the variance of RPE means for the postures. PMID- 11936828 TI - Temporal and force characteristics of fast double-finger, single-finger and hand tapping. AB - The purposes of this study were: (1) to provide descriptive temporal and force data for fast alternate tapping by the index and middle fingers ('double-finger mode'), and to compare it with one finger fast tapping ('single-finger mode'); (2) to determine any differences in the dynamic motor function of individual fingers with these tapping tasks; and (3) to determine any differences between the single-finger mode and tapping with the whole hand ('hand mode'). Eleven healthy males tapped force transducer(s) for 7 s as fast as possible using their dominant hand. The double-finger mode had a greater peak force and a 50% faster tapping frequency than the single-finger mode. There was no correlation between the single-finger and double-finger modes in the temporal data. Tapping with two fingers thus seems to be organized with a motor strategy that is different from that with one finger, and can possibly capture an individual's additional or different motor function. A comparison between the two fingers revealed that the index finger had a significantly faster tapping frequency, indicating the superiority of the index finger over the middle finger in terms of dynamic motor function. The hand mode had a significantly faster tapping frequency with greater peak force than the single-finger mode. A distinction is essential between these two modes. PMID- 11936829 TI - Adolescent versus adult responses to vertical spinal loading. AB - This study compared the vertical spinal creep response between adolescent and adult males. Thirty healthy male subjects, 15 adolescents (aged 12 to 16 years) and 15 adults (aged 30 to 57 years) were measured. Spinal creep was measured continuously over 25 min under loaded and unloaded conditions using a seated stadiometer that controlled spinal posture. Both adolescent and adult subjects showed significant increase in vertical spinal creep with time but the magnitude of vertical spinal creep was significantly greater in adolescent than in the adult subjects. PMID- 11936830 TI - Characterizing human hand prehensile strength by force and moment wrench. AB - Characterizing human hand capabilities or demand created by various occupational tasks or activities of daily living has been mainly accomplished by measuring the maximum force exerted on a force dynamometer in a number of standard grips, for example power, key pinch and tip pinch grips. A framework is proposed instead to characterize human hand prehensile strength in generic form by describing external force and moment wrench capability, where a wrench is a vector describing the forces and moments applied at a point. It is further suggested that if tools and activities are characterized by the internal forces and external forces and moments required, a better understanding of the human prehension in occupational settings and during activities of daily living can be obtained. An example of using a pistol grip drill is used to show the utility of the approach. PMID- 11936831 TI - The influence of moving auditory stimuli on standing balance in healthy young adults and the elderly. AB - The maintenance of postural balance depends on effective and efficient feedback from various sensory inputs. The importance of auditory inputs in this respect is not, as yet, fully understood. The purpose of this study was to analyse how the moving auditory stimuli could affect the standing balance in healthy adults of different ages. The participants of the study were 12 healthy volunteers, who were divided into two age categories: the young group (mean = 21.9 years) and the elderly group (mean = 68.9 years). The instrument used for evaluation of standing balance was a force plate for measuring body sway parameters. The toe pressure was measured using the F-scan Tactile Sensor System. The moving auditory stimulus produced a white-noise sound and binaural cue using the Beachtron Affordable 3D Audio system. The moving auditory stimulus conditions were employed by having the sound come from the right to left or vice versa at the height of the participant's ears. Participants were asked to stand on the force plate in the Romberg position for 20 s with either eyes opened or eyes closed for analysing the effect of visual input. Simultaneously, all participants tried to remain in the standing position with and without auditory stimulation that the participants heard from the headphone. In addition, the variables of body sway were measured under four conditions for analysing the effect of decreased tactile sensation of toes and feet soles: standing on the normal surface (NS) or soft surface (SS) with and without auditory stimulation. The participants were asked to stand in a total of eight conditions. The results showed that the lateral body sway of the elderly group was more influenced than that of the young group by the lateral moving auditory stimulation. The analysis of toe pressure indicated that all participants used their left feet more than their right feet to maintain balance. Moreover, the elderly had the tendency to be stabilized mainly by use of their heels. The young group were mainly stabilized by the toes of their feet. The results suggest that the elderly may need a more appropriate stimulus of tactile and auditory sense as a feedback system than the young for maintaining and control of their standing postures. PMID- 11936832 TI - Glycogen levels and glycogen catabolism in livers from arthritic rats. AB - Hepatic glycogen catabolism and glycogen levels in rats with chronic arthritis were investigated. At 9:00 a.m., the hepatic glycogen contents of ad libitum fed arthritic and normal rats were 225.5+/-17.7 and 332.1+/-28.6 micromol glucosyl units x (g liver)(-1), respectively. Food intake of arthritic and normal rats was equal to 100.1+/-6.7 and 105.0+/-3.1 mg x (g body w)(-1) x (per 24 h)(-1), respectively. In isolated perfused livers from normal and arthritic rats the rates of glucose, lactate and pyruvate release were the same when substrate- and hormone-free perfusion was performed. During an infusion period of 20 min glucagon caused an increment in glucose release of 35.3+/-4.7 micromol x (g liver)(-1) in livers from arthritic rats; in the normal condition the corresponding increment was 69.6+/-5.7 micromol x (g liver)(-1). Lactate and pyruvate productions (indicators of glycolysis) were diminished by glucagon in livers from normal rats; in the arthritic condition an initial stimulation was found, followed by a slow decay, which did not result in significant inhibition at the end of the glucagon infusion period (20 min). The actions of cAMP and dibutyryl-cAMP were similar to those of glucagon. It was concluded that livers from arthritic rats show an impaired capacity of releasing glucose under the stimulus of glucagon. This can be partly due to the lower glycogen levels and partly to a smaller capacity of inhibiting glycolysis. Reduction in glycogen levels was not associated with reduction in food intake or failure in the energetic state of the hepatic cells. These changes in glycogen metabolism may be related to reduced gluconeogenic capacity of the livers and/or to production of inflammatory mediators observed in the arthritis disease. PMID- 11936833 TI - Activation of choline acetyl transferase by dihydrolipoic acid. AB - In a previous article we present results that demonstrated that the enzyme, choline acetyl transferase, was strongly activated by dihydrolipoic acid and that the oxidized form of this substance, lipoic acid itself, was an inhibitor of the enzyme and overcame the stimulatory effects of reduced lipoic acid. The experiments presented in this article show that dialysis of a partially purified preparation of choline acetyl transferase causes complete disappearance of enzyme activity and that addition of dihydrolipoic acid restores activity towards normal. In addition we present experiments with extracts of rat brain and heart as well as rabbit bladder tissue. In these extracts dihydrolipoic acid strongly activates the enzyme. Dialysis of brain and heart extracts causes loss of activity with partial restoration of activity by addition of dihydrolipoic acid. Reduced glutathione has no ability to stimulate activity of the enzyme. We conclude that the results of these experiments strongly support the view that dihydrolipoic acid acts as a coenzyme in the choline acetyl transferase reaction. PMID- 11936834 TI - Changes in the antioxidant content of mononuclear leukocytes from mice with endotoxin-induced oxidative stress. AB - Oxidative stress, associated with a high production of reactive oxygen species (ROS) by immune cells, is involved in the endotoxic shock caused by endotoxin. This oxidative stress is linked to the inability of the immune cells to maintain adequate levels of antioxidants with free radical-scavenging action. Glutathione (GSH) and ascorbic acid (AA) are intracellular and extracellular antioxidants (ROS scavengers) that improve the leukocyte functions. Therefore, in the present work we have determined the reduced GSH and AA content in axillary nodes, spleen, thymus and peritoneal mononuclear leukocytes from BALB/c mice subjected to lethal endotoxic shock produced by intraperitoneal injection of E. coli lipopolysaccharide (LPS, 100 mg/kg), at several times (0, 2, 4, 12 and 24 h) after LPS injection. Endotoxic shock decreased the levels of AA in the leukocytes from the three organs as well as the levels of GSH in axillary nodes and spleen cells while it increased the GSH levels in thymus and peritoneum. These results are in agreement with the oxidative stress and the altered function previously observed in those leukocytes, and they suggest that antioxidant administration may be useful for the treatment of endotoxic shock and other oxidative stress situations with altered immunological responses. PMID- 11936835 TI - alpha-Crystallin chaperone function in diabetic rat and human lenses. AB - This study focussed on the effect of diabetes on the chaperone function of alpha crystallin. The authors relied on diabetic rats with a wide range of plasma glucose levels and non-diabetic control rats to establish a possible relationship between severity of diabetes and alpha-crystallin chaperone activity. In addition, 52-56 and 63-69 year-old diabetic and non-diabetic human lenses were used to show whether diabetes affects alpha-crystallin chaperone activity in human lenses. Correlation between plasma glucose levels and loss of chaperone activity of the alphaL-crystallin fraction in diabetic rats indicated good correlation. The glycemic threshold, reported before for cataract development in diabetic rats, seems to be valid for the chaperone activity loss as well. Analysis of the human lens alphaL-crystallin showed lower chaperone activity in all the diabetic lenses than in the age-matched control lenses. In the 63-69 age group, the loss in chaperone activity due to diabetes was significantly larger than in the 52-56 age group suggesting a dominant effect of duration of diabetes. PMID- 11936836 TI - Cloning, functional expression in Escherichia coli and primary characterization of a new Na+/H+ antiporter, NhaD, of Vibrio cholerae. AB - Vibrio cholerae is the infectious agent of the deadly diarrheal disease, cholera. Na+ ion homeostasis is believed to play a key role in both physiology and pathogenicity of this bacterium. However, molecular mechanisms of sodium exchange in V. cholerae are still poorly understood. In the present work a gene encoding an unusual Na+/H+ antiporter, nhaD, was identified in the V. cholerae genome. nhaD was cloned from Vibrio cholerae and expressed in Escherichia coli. The antiporter functioned in an E. coli nhaAnhaB mutant strain to confer resistance to LiCl and NaCl. When assayed in inside-out subbacterial vesicles, V. cholerae NhaD demonstrated high affinity for Na+ ions (1.1 mM Na+ was required for the half-maximal response at the pH-optimum). The most striking feature of Vc-NhaD is a unique pH-profile of its activity with a sharp maximum at pH 8.0, different from that of any bacterial sodium-proton antiporter described so far. The difference is rationalized as being the result of a His to Arg substitution in a putative pH sensing residue. PMID- 11936837 TI - Bis(maltolato)oxovanadium(IV) inhibits the activity of PTP1B in Zucker rat skeletal muscle in vivo. AB - The insulin signalling pathway consists of a series of phosphorylation and dephosphorylation steps inside the target cell. Phosphotyrosine phosphatase 1B (PTP1B) dephosphorylates phosphotyrosine (pTyr) residues present on the insulin receptor (IR). In this study we examined the effect of bis(maltolato)oxovanadium(IV) (BMOV) on PTP1B and its possible role in the amelioration of insulin resistance. Fourteen to sixteen week old fatty Zucker rats (F), an animal model of insulin resistance, were treated with BMOV in drinking water for 3 weeks (FT) along with age matched lean littermate controls. The fatty rats responded to vanadium with a significant decrease in plasma insulin, (F = 5.1+/-0.8 FT = 3.3+/-0.7 ng/ml). During insulin resistance the activity of PTP1B has been shown to increase, thus diminishing insulin signalling in the target tissues. Hence, PTP1B is an important target for anti-diabetic drug research. In our investigation we found that the PTP1B activity was increased to 200% in the skeletal muscle of untreated Zucker fatty rats compared to lean littermates. Three weeks of BMOV treatment reduced the activity of PTP1B by 25% in fatty treated rats, in vivo, compared to untreated fatty rats. There was no significant change in the activity of PTP1B in the lean treated rats. There was also no difference in the gene expression of PTP1B in the skeletal muscle of different groups of rats. Vanadium compounds also inhibited PTP1B in vitro. These results indicate that PTP1B may be a potential target for the action of BMOV at least in the Zucker fatty rat model. PMID- 11936838 TI - Inhibition of MCP-1 and MIP-2 transcription and translation by mimosine in muscle tissue infected with the parasite Trichinella spiralis. AB - Mimosine is a non-toxic plant aminoacid which is an effective inhibitor of DNA replication by acting at the S-phase. In this study we infected mice with T. spiralis, a nematode parasite, and studied the inflammatory response through the determination of MIP-2, a C-X-C chemokine and MCP-1, a C-C chemokine in the inflamed area around the parasitic cyst. The animals were infected and their diaphragms were tested for inflammatory response. MCP-1 and MIP-2 was tested after 1, 10, 20, 30, and 40 days post inoculation, before and after mimosine treatment. The inflammatory index was calculated by counting the white blood cells around the nematode cysts, while expression of MIP-2 and MCP-1 was calculated by ELISA method and transcription by Northern blot and RT-PCR. Here we found that mimosine strongly inhibited the inflammatory index in the diaphragmatic tissue at 10, 20, 30 and 40 days post-treatment. In these experiments, mimosine had no effect on the number of cysts produced. In addition, we found that MCP-1 transcription and translation was completely inhibited by mimosine, while MIP-2 transcription and translation was partially inhibited at 30 and 40 days; yet it was totally inhibited after 10 and 20 days in encysted diaphragm tissue infected by T. spiralis. Our studies suggest that mimosine has an inhibitory effect through the inhibition of cytoplasmatic serine hydroxymethyltransferase altering the cell cycle of white blood cells. This study suggests for the first time the premise that mimosine acts as an anti inflammatory compound. PMID- 11936839 TI - Heat shock and ceramide have different apoptotic pathways in radiation induced fibrosarcoma (RIF) cells. AB - Heat shock induces various cellular responses including inhibition of protein synthesis, production of heat shock proteins (HSPs) and induction of thermotolerance. The molecular mechanisms of the processes have not been well understood. It has been proposed that ceramide formation during heat shock mediates heat shock induced apoptosis. We examined whether C2-ceramide mimicked the cellular response to heat shock in RIF-1 cells and their thermotolerant derivative TR-RIF-1 cells. Discernible effects between heat shock and C2-ceramide treatments were observed in cellular changes such as total protein synthesis, HSP synthesis, stress-activated protein kinase/c-Jun N-terminal kinase (SAPK/JNK) activity and PARP cleavage. Heat shock immediately inhibited cellular protein synthesis, which was recovered by synthesizing HSPs first and then whole proteins later. Heat shock also activated SAPK/JNK and increased PARP cleavage in dose dependent manner. Thermotolerant TR-RIF-1 cells responded to heat shock more insensitively than RIF-1 cells. On the other hand, C2-ceramide treatment did not accompany any changes induced by heat shock. No discernible differences between RIF-1 and TR-RIF-1 cells were observed by C2-ceramide treatment. We tried to figure out how C2-ceramide interacts with cellular membrane and found that exogenous C2-ceramide was incorporated into the outer monolayer and flipped into the inner monolayer of human erythrocytes in ATP-dependent manner. However, the rate of C2-ceramide incorporation was similar in control and thermotolerant cells. In summary, thermotolerant cells are resistant to heat shock induced apoptotic signaling but not resistant, rather sensitive to membrane disturbing C2 ceramide mediated apoptosis. These results suggest that heat shock and ceramide have different signal transduction pathways. PMID- 11936840 TI - Passive cigarette smoke and the renal glyoxalase system. AB - Cigarette smoking is associated with a number of fatal diseases, including cancer of different organs. A number of oxoaldehydes are found in cigarette smoke, among which methylglyoxal (MG) is known to cause toxicity to cells upon accumulation. In biological systems, MG is converted to s-d-lactoylglutathione by glyoxalase I with reduced glutathine (GSH) as a cofactor, and s-d-lactoylglutathione is converted to D-lactic acid with simultaneous regeneration of GSH, by glyoxalase II. In the present study, we have investigated the status of the glyoxalase enzymes in kidney tissues from rats exposed to passive cigarette smoke. No significant change has been noted in glyoxalase I activity. Glyoxalase II was decreased during 1 and 2 weeks of exposure, and after that the activity was increased. The initial decrease in the activity of gly II may be due to the excess amount of methylglyoxal generated due to smoke exposure or the adduct formed by MG and GSH which known to inhibit gly II activity. Both enzymes help in the detoxification of cigarette smoke induced chemicals and biochemicals. PMID- 11936841 TI - Suppressive effect of endogenous regucalcin on deoxyribonuclic acid synthesis in the nuclei of rat renal cortex. AB - The effect of regucalcin, a regulatory protein of Ca2+ signaling, on deoxyribonucleic acid (DNA) synthesis activity in the nuclei isolated from rat renal cortex was investigated. The addition of calcium chloride (10-100 microM) in the reaction mixture containing the nuclei caused a significant decrease in DNA synthesis activity. Nuclear DNA synthesis activity was significantly raised in the presence of EGTA (1 mM), a chelator of Ca2+, indicating that nuclear Ca2+ has an inhibitory effect. Regucalcin (0.1-0.5 microM) added in the reaction mixture in the presence of either EGTA (1 mM) or calcium chloride (50 microM) had a significant inhibitory effect on nuclear DNA synthesis activity. The presence of anti-regucalcin monoclonal antibody (10-50 ng/ml) in the reaction mixture caused a significant increase in DNA synthesis activity. This increase was completely abolished by the addition of regucalcin (0.5 microM). The effect of anti-regucalcin monoclonal antibody in increasing DNA synthesis was enhanced in the presence of EGTA. Additionally, an inhibitory effect of calcium chloride (10 or 50 microM) was enhanced in the presence of anti-regucalcin monoclonal antibody (25 ng/ml). The present study demonstrates that endogenous regucalcin has a suppressive effect on DNA synthesis in the nuclei of rat renal cortex. PMID- 11936842 TI - Inhibitory effects of extracellular Mg2+ on intracellular Ca2+ dynamic changes and thapsigargin-induced apoptosis in human cancer MCF7 cells. AB - The effects of extracellular Mg2+ on both dynamic changes of [Ca2+]i and apoptosis rate were analysed. The consequences of spatial and temporal dynamic changes of intracellular Ca2+ on apoptosis, in thapsigargin- and the calcium ionophore 4BrA23187-treated MCF7 cells were first determined. Both 4BrA23187 and thapsigargin induced an instant increase of intracellular Ca2+ concentrations ([Ca2+]i) which remained quite elevated (> 150 nM) and lasted for several hours. [Ca2+]i increases were equivalent in the cytosol and the nucleus. The treatments that induced apoptosis in MCF7 cells were systematically associated with high and sustained [Ca2+]i (150 nM) for several hours. The initial [Ca2+]i increase was not determinant in the events triggering apoptosis. Thapsigargin-mediated apoptosis and [Ca2+]i rise were abrogated when cells were pretreated with the calcium chelator BAPTA. The role of the extracellular Mg2+ concentration has been studied in thapsigargin treated cells. High (10 mM) extracellular Mg2+, caused an increase in basal [Mg2+]i from 0.8+/-0.3 to 1.6+/-0.5 mM. As compared to 1.4 mM extracellular Mg2+, 1 microM thapsigargin induces, in 10 mM Mg2+, a reduced percentage from 22 to 11% of fragmented nuclei, a lower sustained [Ca2+]i and a lower Ca2+ influx through the plasma membrane. In conclusion, the cell death induced by thapsigargin was dependent on high and sustained [Ca2+]i which was inhibited by high extracellular and intracellular Mg2+. PMID- 11936843 TI - Inhibition of human low density lipoprotein oxidation by active principles from spices. AB - Spice components and their active principles are potential antioxidants. In this study we examined the effect of phenolic and non-phenolic active principles of common spices on copper ion-induced lipid peroxidation of human low density lipoprotein (LDL) by measuring the formation of thiobarbituric acid reactive substance (TBARS) and relative electrophoretic mobility (REM) of LDL on agarose gel. Curcumin, capsaicin, quercetin, piperine, eugenol and allyl sulfide inhibited the formation of TBARS effectively through out the incubation period of 12 h and decreased the REM of LDL. Spice phenolic active principles viz. curcumin, quercetin and capsaicin at 10 microM produced 40-85% inhibition of LDL oxidation at different time intervals while non-phenolic antioxidant allyl sulfide was less potent in inhibiting oxidation of LDL. However, allyl sulfide, eugenol and ascorbic acid showed pro-oxidant activity at lower concentrations (10 microM) and antioxidant activity at higher concentrations (50 microM) only. Among the spice principles tested quercetin and curcumin showed the highest inhibitory activity while piperine showed least antioxidant activity at equimolar concentration during initiation phase of oxidation of LDL. The inhibitory effect of curcumin, quercetin and capsaicin was comparable to that of BHA, but relatively more potent than ascorbic acid. Further, the effect of curcumin, quercetin, capsaicin and BHA on initiation and propagation phases of LDL oxidation showed that curcumin significantly inhibited both initiation and propagation phases of LDL oxidation, while quercetin was found to be ineffective at propagation phase. These data suggest that the above spice active principles, which constitute about 1-4% of above spices, are effective antioxidants and offer protection against oxidation of human LDL. PMID- 11936844 TI - The impact of insulin-like growth factor-1 on the pattern of cardiac elongation factor-2 variants in a model of overload. AB - Because of its key role in proteosynthesis, the total content of elongation factor-2 (EF-2) and the distribution of six main EF-2 variants were investigated after Pseudomonas Exotoxin A catalyzed [37P]ADP-ribosylation using 1D-PAGE and isoelectric focusing (IEF) in a rat model of hemodynamic overload with variable degrees of cardiac hypertrophy: Chronic NO-synthase inhibition by L-NAME (N-omega nitro-L-arginine-methyl-ester; 0.75 mg/ml drinking water) induced arterial hypertension without hypertrophy but myocardial apoptosis; additional treatment with IGF-1 (osmotic micropumps) did not modify hypertension but reduced apoptosis allowing moderate hypertrophy of the left ventricles. Total EF-2 did not significantly increase in rats with hemodynamic overload with or without IGF-1 supplementation. A positive correlation was found between an acidic EF-2 variant and apoptosis (p = 0.01). Hypertrophy under additional IGF-1 was combined with a shift of the EF-2 variants to basic subtypes (p < 0.01). This finding may be indicative of the trophic potency of IGF-1. PMID- 11936845 TI - Characterization and expression of three novel differentiation-related genes belong to the human NDRG gene family. AB - NDRG1 (N-Myc downstream regulated) is upregulated during cell differentiation, repressed by N-myc and c-myc in embryonic cells, and suppressed in several tumor cells. A nonsense mutation in the NDRG1 gene has been reported to be causative for hereditary motor and sensory neuropathy-Lom (HMSNL), indicating that NDRG1 functions in the peripheral nervous system necessary for axonal survival. Here, we cloned three human cDNAs encoding NDRG2 (371aa), NDRG3 (375aa) and NDRG4 (339aa), which are homologous to NDRG1. These three genes, together with NDRG1, constitute the NDRG gene family. The phylogenetic analysis of the family demonstrated that human NDRG1 and NDRG3 belong to a subfamily, and NDRG2 and NDRG4 to another. At amino acid (aa) level, the four members share 53-65% identity. Each of the four proteins contains an alpha/beta hydrolase fold as in human lysosomal acid lipase. Expression of the fusion proteins NDRG2/GFP, NDRG3/GFP and NDRG4/GFP in COS-7 cells showed that all of them are cytosolic proteins. Based on UniGene cluster analysis, the genes NDRG2, NDRG3 and NDRG4 are located at chromosome 14q11.1-11.2, 20q12-11.23 and 16q21-22.1, respectively. Northern and dot blot analysis shows that all of the three genes are highly expressed in adult brain and almost not detected in the eight human cancer lines. In addition, in contrast to the relatively ubiquitous expression of NDRG1, NDRG2 is highly expressed in adult skeletal muscle and brain, NDRG3 highly expressed in brain and testis, and NDRG4 specifically expressed in brain and heart, suggesting that they might display different specific functions in distinct tissues. PMID- 11936846 TI - Overexpression of the nucleolar protein nucleophosmin/B23 in collagen lattice cultured fibroblasts: potential role in the control of protein synthesis. AB - Fibroblasts cultivated in tridimensional collagen lattices exhibit a downregulation of protein synthesis, related to decreased ribosomal RNA (rRNA) content and half life, when compared to monolayer cultivated cells. The involvement in this process of nucleophosmin/B23, a nucleolar phosphoprotein with ribonuclease properties, was checked. We compared production of nucleophosmin/B23 in monolayer and collagen lattice cultured fibroblasts. A significant increase of nucleophosmin/B23 mRNA levels was noticed in lattice-cultured fibroblasts vs monolayers (+154%, p < 0.05). A concomitant enhancement of nucleolar nucleophosmin/B23 content was found (+112%, p < 0.001). Simultaneously, ribonuclease activity contained in nucleolar extracts from collagen lattice cultured fibroblasts was significantly increased (+54%, p < 0.01). These data demonstrate that extracellular collagen matrix induces the overexpression of nucleophosmin/B23, and suggest that the regulation of protein syntheses in collagen lattice cultures may be explained, at least partly, by an increased degradation of neosynthesized rRNAs dependent on nucleophosmin. PMID- 11936847 TI - The 21-day postnatal rat ventricular cardiac muscle cell in culture as an experimental model to study adult cardiomyocyte gene expression. AB - The purpose of this study was to develop and characterize a cardiomyocyte culture system for use as an experimental model to study the mechanism(s) by which cardiac muscle cells permanently exit the cell cycle during early neonatal life. Ventricular cardiomyocytes, isolated by retrograde perfusion of hearts from 21 day-old and adult rats, were compared through 10 days of culture. Expression patterns of genes encoding developmentally programmed proteins were determined to be similar between cardiomyocytes cultured from 21-day-old and adult rats, using the reverse transcription polymerase chain reaction. A lacZ-expressing reporter gene was used to test the efficiency of gene delivery in cultured cardiomyocytes. Transfections using cationic liposomes yielded 24+/-7, 25+/-7 and 10+/-1% cardiomyocytes positive for beta-galactosidase activity in cultured 1-day, 21-day and adult cardiomyocytes, respectively. Direct needle microinjection resulted in 48+/-7, 35+/-6 and 37+/-5% cardiomyocytes positive for enzymatic activity in 1 day, 21-day and adult cardiomyocytes, respectively. Cell cycle-specific cDNA arrays were used to analyze the expression pattern of cell cycle-related genes in 12-O-tetradecanoyl-phorbol-13-acetate (TPA)- and non-TPA-treated cultured 21-day cardiomyocytes. Based on the similarity of cultured 21-day to adult ventricular cardiomyocytes and their high transfection efficiencies, we propose the use of cultured cardiomyocytes from 21-day-old rat ventricles as an experimental model system for the study of adult cardiomyocyte gene expression and cell cycle machinery. PMID- 11936848 TI - Serum is a rich source of ligands for the scavenger receptor of hepatic sinusoidal endothelial cells. AB - The present study was prompted by findings in our laboratory showing that serum effectively inhibits scavenger receptor (SR)-mediated endocytosis in hepatic sinusoidal endothelial cells (SEC). Experiments with SEC in vitro showed that the presence of 20% human serum inhibited endocytosis of SR ligands, 125I formaldehyde treated bovine serum albumin (FSA) and 125I-nidogen, by 30 and 50%, respectively, whereas pre-heated foetal bovine serum (10%) inhibited endocytosis of 125I-FSA by as much as 56%. Human, bovine and rat serum had similar inhibitory effect on endocytosis in SEC. Fractionation of foetal bovine and human serum on anion exchange chromatography demonstrated that the inhibitory principle co purified with macromolecules of high negative charge. The serum fraction that most effectively inhibited SR-mediated endocytosis of 125I-FSA did not affect mannose receptor-mediated endocytosis of 125I-mannan to the same extent. Trap labelled negatively charged serum fraction administered intravenously to rats was eliminated almost exclusively by liver, with a blood decay of 50% over the first 3 min after injection. Isolation of liver cells showed that the populations of Kupffer cells and SEC contained 39 and 61% of liver radioactivity 30 min after injection of trap-labelled negatively charged fractions prepared from pre-heated ('complement inactivated') foetal bovine sera. These findings suggest that the process of serum formation from native blood generates appreciable amounts of macromolecules that compete specifically with the SR for endocytosis in SEC. The inhibitory power of pre-heated serum is particularly great. For this reason pre heated serum should be used with caution in studies of SR in SEC. PMID- 11936849 TI - The characterization and purification of a human transcription factor modulating the glutathione peroxidase gene in response to oxygen tension. AB - An oxygen responsive transcription factor regulating human glutathione peroxidase gene (GPx) through two oxygen responsive elements (ORE I and ORE2) has been purified and characterized by sequence-specific DNA affinity chromatography. The DNA binding activity, termed Oxygen Responsive Element Binding Protein (OREBP), was partially represented by a 77 kD polypeptide (p70) possessing a blocked N terminus. The p70 subunit co-eluted with an 86 kD subunit (p80) from affinity columns. N-terminal sequencing analysis of the 86 kD component revealed that this protein represented the larger member of the Ku antigen complex. The identity of the purified 77 kD subunit was determined by Western blot analysis using an antibody directed against the p70 protein. In addition to binding the GPx-ORE, the OREBP was itself regulated by oxygen tension. It was found that the abundance of the ORE binding activity was decreased in cells maintained at low oxygen tension (40 mm Hg). Anti-Ku-antibodies specifically supershifted the OREBP-ORE DNA complex. These observations further add to the numerous nuclear roles of the Ku-transcription factor. PMID- 11936850 TI - Green tea catechins decrease apolipoprotein B-100 secretion from HepG2 cells. AB - To understand the hypocholesterolemic activity of green tea, our in vitro studies screened the relative efficacy of two structurally distinct green tea catechins, epicatechin (EC) and epigallocatechin gallate (EGCG), on apolipoprotein B-100 (apoB) and lipid production using a well established human hepatoma cell-line, HepG2, as the model system. This study showed that HepG2 cells pretreated with EC and EGCG for 8 h exerted a dose-dependent inhibitory effect on apoB secretion. Total protein and albumin synthesis and secretion were unaffected indicating the effects on apoB secretion to be specific. Under lipid-rich conditions, apoB secretion was markedly reduced by EGCG and to a lesser extent by EC at 50 microM. Mechanistic study showed that tea catechins inhibited apoB secretion via a proteasome-independent pathway as indicated by a lack of response to N-acetyl leucyl-leucyl-norleucinal (ALLN), a proteasome inhibitor. The effect on apoB secretion was also found to be independent of lipid biosynthesis. In summary, the data suggest that EGCG in contrast to EC is a potent inhibitor of apoB secretion. The results indicate that the gallate moiety in the catechin molecule may result in a beneficial effect on lipid metabolism in terms of apoB secretion. PMID- 11936851 TI - A novel peptide derivative exhibits anti inflammatory and antioxidant activity in adjuvant induced arthritis in rats. AB - A tetrapeptide derivative PEP1261 [Boc-Lys-(Boc)-Arg-Asp-Ser-(tBu)-OtBu], corresponding to residues 39-42 of human lactoferrin, was tested for its antiinflammatory action in adjuvant induced arthritis in rats. Administration of heat killed Mycobacterium tuberculosis (500 microg/0.1 ml of paraffin oil) intradermally into the foot pad of right hind paw resulted in an increased paw volume and an increase in the levels of reactive oxygen species and beta glucuronidase as well as a decrease in the antioxidants levels. PEP1261, at an effective dose of 10 mg/kg body wt., exhibited a significant antiarthritic activity as evidenced by lowering of paw volume and inhibited the free radicals toxicity by increasing the antioxidants levels. This peptide derivative was also shown to have a membrane stabilizing action by significantly decreasing the total and free activity of beta-glucuronidase and inhibiting the rate of release of the enzyme from lysosomal rich fraction. Histopathological studies confirmed the above results by showing a decrease in mononuclear cell infiltration, hypertrophy, hyperplasia and pannus formation after PEP1261 treatment in arthritic rats. PMID- 11936853 TI - Artificial inseminations with frozen semen and application of DNA fingerprint for parents/offspring conformation on Great Pyrenees. AB - Frozen semen of Great Pyrenees, air-shipped from France, was artificially inseminated (AI) to two Great Pyrenees bitches. Blood LH and P concentrations and vulval width principally determined optimal breeding time of bitches. AI were performed on 17 to 21 days from the onset of estrous bleeding, and 25 days after final Al, pregnancy of one bitch, with two fetuses, was identified by ultrasound image diagnostic equipment. On 62 days, the bitch whelped one male pup and one female pup. Blood samples were obtained from four parents and offspring, and their DNA was extracted. DNA samples digested with four kinds of restriction endonucleases, and resulting fragmented DNA were used as test samples for DNA fingerprinting with Myo prove. As a result bands from pups, displayed by any enzyme, were inherited from either parent, and clear parent-offspring relationships were identified. PMID- 11936852 TI - Variations in biochemical and pharmacological properties of Indian cobra (Naja naja naja) venom due to geographical distribution. AB - Indian cobra (Naja naja naja) venom obtained from three different geographical regions was studied in terms of electrophoretic pattern, biochemical and pharmacological activities. SDS-PAGE banding pattern revealed significant variation in the protein constituents of the three regional venoms. The eastern venom showed highest indirect hemolysis and hyaluronidase activity. In contrast, western and southern venoms were rich in proteolytic activity. All the three regional venoms were devoid of p-tosyl-L-arginine methyl ester hydrolysing activity. The eastern venom was found to be most lethal among the three regional venoms. The lethal potency varied as eastern > western > southern regional venoms. In addition, all the three regional venoms showed marked variations in their ability to induce symptoms/signs of neurotoxicity, myotoxicity, edema and effect on plasma coagulation process. Polyclonal antiserum prepared against the venom of eastern region cross-reacted with both southern and western regional venoms, but varied in the extent of cross-reactivity by ouchterlony immunodiffusion and ELISA. PMID- 11936854 TI - Morphological characteristics by SEM observations and regulatory volume decrease (RVD) of tentacular nematocytes isolated by heat dissociation from Aiptasia diaphana (Cnidaria: Anthozoa). AB - This paper examines the morphological characteristics and response to hyposmotic shock of nematocytes isolated by heat dissociation at 45 degrees C for 20 min from the tentacles of Aiptasia diaphana, an anthozoan living in the brackish water of Lake Faro (Messina, Italy). Morphological characteristics were evaluated by scanning electron microscopy (SEM) observations and cytological test, functional characteristics by exposure to 35% hyposmotic stress. 81% of the population of isolated nematocytes had ciliary protrusions. Microbasic mastigophore and amastigophore nematocytes had a hair bundle at the crown shaped apex composed of ciliary protrusions of different heights and diameters. In basitrichs, instead, a single ciliary protrusion was observed. Following exposure to 35% hyposmotic shock, nematocytes isolated by heat dissociation did not show RVD, while, following treatment with 1 microM and 2 microM gramicidin-S, activation of volume regulation in conditions of hyposmoticity was observed. The effect of gramicidin was concentration-dependent and confirmed the relevant role of conductance to K+ in volume regulation, as previously described (17) on nematocytes isolated by chemical methods. PMID- 11936855 TI - Inhibition of proliferation of human lymphoma cells U937 by a 50 Hz electromagnetic field. AB - Weak pulsating electromagnetically induced fields (PEMF) by Helmholtz coils changes cell metabolism, if cells are treated with a certain range of frequency and amplitude. The influence on proliferation of human histiocytic lymphoma cells U937 has been studied applying a sinusoidal 50 Hz field with amplitudes of the flux density B = 0.3 to 4.7 mT for 4 days. No difference between experiment and control was found in the region 0.3 and 2 mT. However, stronger fields (> or =2.5 mT) inhibit cell division. Fields > or =3.5 mT treatment kill > or =80% of the cell number at the beginning (1.5 x 10(5)/ml). This effect may be an electromagnetocally induced cell death as the first step for a non-invasive application on cell proliferation process. PMID- 11936856 TI - An active fraction of unfractionated heparin from a natural source is recognized by the first component of the complement system. AB - Heparin is a natural glycosaminoglycan with chains consisting of alternating 1-4 linked residues of sulphated uronic acids (L-iduronic in great proportion) and D- glucosamine attached to a serine-glycine linear protein core. In our previous experiments with a low molecular weight heparin (Mr = 4.000 to 6.000) obtained by partial chemical degradation of the original heparin we could concentrate its anticoagulant activity by precipitation with the first component of the human complement system. In order to confirm these results with a more physiological unfractionated heparin we used commercial heparin from porcine intestinal mucose with a high molecular weight (9.000-15.000) and a specific activity of 179 U/mg. An heparin fraction with high anticoagulant activity was isolated from the precipitate of the interaction between this high molecular weight heparin isolated from a natural source, and the first component of the human complement system. Our results confirmed, in opposition to almost all early literature, that under very strict conditions of pH 6.0, calcium chloride concentration (2 mM), and very low ionic strength (25 mM), the first component of the human complement cascade recognize heparin fractions "enriched" in the high affinity sequence for the antithrombin III. PMID- 11936857 TI - Distribution of tissue kallikrein in the integumental blood vessel system of wild mammals. AB - The study demonstrates tissue kallikrein with the help of a very sensitive immunohistochemical B-SA method in smooth muscle cells and myocytes / pericytes of the wall of subepidermal arterioles and capillaries of 17 mammalian species from 6 different orders. Positive immunoreactions could also be detected in the wall of blood vessels near to and between hair follicles and skin glands. The reaction staining in subepidermal blood vessels was particularly distinct in animals with a vital epidermis that was significantly thicker than 20 microm. The results obtained are discussed with special regard to the general skin biology of the wild mammals studied, considering comparative hair coat density and normal hazards to the integument. PMID- 11936858 TI - Femtosecond near-infrared laser pulse induced strand breaks in mammalian cells. AB - Multiphoton laser scanning microscopy (MPLSM) is based on non-resonant simultaneous absorption of two or three near infrared (NIR) photons by a fluorophore in the subfemtoliter focal volume of a high numerical aperture (N.A. 1.3) objective. The higher penetration depth of NIR radiation enables optical sectioning across thick biological specimens and because of the absence of efficient single photon absorbers in the NIR spectral region of 700 to 1200 nm there is hardly any out-of-focus photodamage and photobleaching. Recent in vitro studies (14) have demonstrated that irradiation of supercoiled plasmid DNA with intense multiphoton NIR 810 nm of 140 fs pulse width, 76 MHz pulse repetition rate results in single strand breaks as a result of simultaneous absorption of three or more photons. Herein, we have investigated the influence of 800 nm NIR 170 fs laser pulses, 80 MHz pulse repetition frequency at mean powers of 2 to 20 mW on nuclear DNA of unlabelled PtK2 cells. In situ TdT-mediated dUTP-nick end labelling (TUNEL) revealed that cells exposed to the NIR irradiation above > or =5 mW mean laser power alone contained TUNEL-positive nuclei. The intensity of TUNEL fluorescence was relatively higher at increased mean NIR laser power. These results provide evidence that DNA strand breaks also occur in vivo when mammalian cells are exposed to high average power > or =5 mW NIR irradiation during MPLSM possibly due to multiphoton absorption process. Because intense DNA fragmentation is one of the hall marks of programmed cell death it is hypothesised that NIR induced cell death is by apoptosis. PMID- 11936859 TI - Regional difference in muscle fiber type and glucose uptake of mouse gastrocnemius at rest. AB - Regional differences in muscle fiber types and their glucose uptake in superficial, middle and deep regions of mouse gastrocnemius at rest were studied. The enzyme activities of succinate dehydrogenase (SDH), alpha-glycerophosphate dehydrogenase (alpha-GPDH) and myofibrillar ATPase were used for the classification of each muscle fiber type. For the evaluations of glucose uptake, 2-deoxyglucose (2-DG) microradioautography and expression of the glucose transporter 4 (GLUT4) protein were applied. In superficial region of mouse gastrocnemius, all fibers were IIx or IIb and were low for 2-DG uptake and GLUT4 expression. They were anaerobic and fast twitch. In middle region, fibers showing low 2-DG uptake and small amounts of GLUT-4 protein (IIx and IIb) dominated (80.4%). Most fibers were anaerobic and fast twitch. Others were type IIc and IIa. In deep region, most fibers (86.9%) showed high 2-DG uptake and large amounts of GLUT4 protein (I, IIc and IIa). They were oxidative and slow twitch. PMID- 11936860 TI - Disturbed copper transport in humans. Part 1: mutations of the ATP7A gene lead to Menkes disease and occipital horn syndrome. AB - Mutations of the ATP7A gene (OMIM 300011) lead to the Menkes disease (MD, OMIM 309400) involving impaired brain development, neurological degeneration, connective tissue abnormalities, and high lethality in early infancy. Occipital horn syndrome (OHS, OMIM 304150), a milder phenotype, is also caused by ATP7A gene mutations. In MD patients, an early copper-histidine treatment may prevent the neurological impairment and prolong survival leading to an OHS phenotype. To demonstrate the genotype/phenotype correlation, two male patients are reported with different ATP7A gene mutations and several phenotypes. In the first patient with the MD phenotype, a mutation within the exon 20 (Gln1288Ter) was found producing a stop codon just prior to the highly conserved ATP binding domain. The OHS phenotype of the second patient was caused by a splice site mutation involving the position +6 of intron 6 within a copper binding domain. Small amounts of correctly spliced ATP7A transcript were sufficient to develop the milder OHS phenotype in this patient (OMIM 30001.0006). In conclusion, mutations of the copper transporting P-type ATPase ATP7A gene cause distinct human diseases showing some genotype/phenotype correlation and implications for treatment. PMID- 11936861 TI - Disturbed copper transport in humans. Part 2: mutations of the ATP7B gene lead to Wilson disease (WD). AB - Mutations in the Wilson disease gene ATP7B, a P-type ATPase, are responsible for copper accumulation in the liver and other organs leading to Wilson disease (WD, OMIM 277900). Clinical manifestations of Wilson disease (WD) include chronic liver disease, acute hepatic failure or neuropsychiatric diseases. Since potent medical treatments are available to prevent disabling residual symptoms, early diagnosis is crucial. To demonstrate the clinical course and genetic findings, a male patient with a novel mutation in the ATP7B gene, a 10 base pair insertion in exon 6 (1927ins 10), and a second missense mutation in exon 13 (P992L) is reported. The patient presented with signs of chronic liver disease at the age of 10 years. Clinical findings included hepatomegaly, elevated liver enzymes and coagulopathy. A combination treatment with the copper chelating agent D penicillamine and zinc acetate was started leading to normalization of liver function and no appearance of neurological signs or Kayser-Fleischer ring after 7 years follow-up. Truncating mutations of the ATP7B gene (insertions, deletions, nonsense mutations) leading to gross loss of C-terminal parts of the protein, thereby probably completely destroying the protein function, may correlate with a hepatic phenotype and early onset as seen in the patient presented. PMID- 11936862 TI - Loss of cardiac contractile response to tetrahydropapaveroline with advanced age and hypertension. AB - Tetrahydropapaveroline (THP), a condensation product of ethanol-derived acetaldehyde, potentiates cardiac function through beta-adrenoceptor. We have recently shown that THP-induced cardiac contractile action is likely due to its action at the single myocyte level, and is markedly diminished during early hypertension. Cardiac function alters with advanced age reminiscent of hypertension. This study was to examine cardiac contractile response to THP with advanced age and hypertension. Left ventricular papillary muscles and myocytes were isolated from normotensive (WKY) or hypertensive (SHR) rats, and stimulated to contract at 0.5 Hz. Mechanical parameters evaluated include: peak tension developed (PTD)/peak shortening (PS), time-to-PTD/PS (TPT/TPS), time-to-90% relaxation/relengthening (RT90/TR90), and maximal velocities of contraction/relaxation (+/- VT/+/- dLdt). Intracellular Ca2+ transients were measured as fura-2 fluorescence intensity changes (AFFI). THP (0.1-100 microM) increased PTD in 10- but not 36-wk-old WKY rat myocardium. THP elicited positive, negative or no response on PS in myocytes from 10-wk WKY, 36-wk WKY, and 36-wk SHR groups, respectively. Interestingly, THP elicited discrepant response on intracellular Ca2+ transient compared with that of myocyte shortening. THP increased AFFI in 10-wk WKY and 36-wk SHR myocytes while exhibiting a significant inhibiting action in 36-wk WKY myocytes. Lastly, THP shortened TPT/TPS, RT90/TR90 and increased +VT in all animal groups. These results indicate that the THP induced myocardial contractile response is altered in advanced age and hypertension, in a manner similar to early stage of hypertension. It is possible that altered intracellular Ca2+ responsiveness may be involved in THP-induced action. PMID- 11936863 TI - FT-IR microscopic characterization of normal and malignant human colonic tissues. AB - Fourier-transform infrared spectroscopy (FT-IR) employs a unique approach to optical diagnosis of tissue pathology based on the characteristic molecular vibrational spectra of the tissue. In this study, we report infrared absorption spectra of formalin fixed, paraffin embedded normal and malignant human colonic tissues from ten different patients. Our method is based on microscopic infrared study (FT-IR-microscopy) of thin tissue specimens in parallel with normal histopathological analysis, which serves as a reference. Our results indicate that the normal colonic tissue has a stronger absorption than the cancerous type over a wide region in all ten cases. The detailed analysis showed that there is a significant decrease in carbohydrate levels, total phosphate and also possibly creatine contents for cancerous tissue in comparison to the controls. Also, RNA/DNA ratio increased in cancerous tissues relative to the normals in all the patients. The results of Linear Discriminant Analysis (LDA) showed that the normal and malignant cells could be identified with about 89% accuracy. PMID- 11936864 TI - Hepatitis C virus RNA detection by in situ RT-PCR in formalin-fixed paraffin embedded liver tissue. Comparison with serum and tissue results. AB - The purpose of this study was to localize HCV RNA in formalin-fixed paraffin embedded liver biopsies of 15 patients with chronic hepatitis C using in situ RT PCR method. The results were compared to serum and tissue extract analysis of HCV RNA. HCV RNA was detected in 80% of the sera tested, in 40% of the corresponding hepatic tissue extract and in 60% of the tissue sections tested by in situ RT PCR. Compared to the serum positive cases, 67% of the cases were positive with in situ RT-PCR and 41% were positive with tissue extract detection. 50% of the cases in situ RT-PCR positive were also positive with tissue extract detection. These results underlined the complementarity of the different methods of viral detection for the precise diagnosis of hepatitis C. PMID- 11936865 TI - Fatty acid ethyl esters and ethanol-induced pancreatitis. AB - Conjugation of xenobiotic alcohols with endogenous fatty acids is considered one of the mechanisms of their retention in the target organs. A number of fatty acid esters of alcohol's detected in the human tissues were found to be toxic in vivo and in vitro. Non-oxidative metabolism of ethanol resulting in the formation of fatty acid ethyl esters (FAEEs) appears to be one of the major pathways of ethanol disposition in the pancreas during chronic alcohol abuse, and could be associated with pancreatitis. In most cases, pancreatic damage occurs in alcoholics preceding the onset of clinical pancreatitis. Early markers of ethanol induced pancreatitis could be important for early prevention of such injury. Although FAEEs have been implicated in the ethanol-induced pancreatitis, mechanism(s) of such injury is not well understood. Studies by others and by our own group have shown that plasma levels of FAEEs correlate well with plasma/blood alcohol concentration. FAEE synthase is known to catalyze the formation of FAEEs. The activity of FAEE synthase was found highest in the pancreas. Excessive synthesis of FAEEs during chronic alcohol abuse in the pancreas may be associated with pancreatic injury as supported by in vivo and cell culture studies. Human studies correlating plasma FAEE levels with that of markers of pancreatic injury could be important in developing markers of ethanol-induced toxicity. Although toxicity of exogenously administered FAEEs is shown in vivo and in vitro, the toxicity associated with endogenously formed FAEEs has not been studied. Therefore, studies regarding the role of endogenously formed FAEEs could be important in understanding the mechanism of ethanol-induced pancreatitis. PMID- 11936866 TI - Use of microsatellite markers for parentage testing of two litters of beagles born after artifcial insemination with mixed fresh semen. AB - This study was undertaken to determine if 20 kinds of microsatellite markers, which have already been reported, were useful for the parentage testing of 16 Beagle puppies born after artificial insemination using intentionally mixed semen from six male Beagles. Of the 20 kinds of markers used in this study, 17 kinds (the exceptions were CPH6, FH0020 and FH0419) produced 3 to 5 kinds of alleles, and using these markers, the parentage was finally established for all of the 16 puppies. These 17 kinds of markers showed sufficient amplification in PCR and had no non-specific PCR products or alleles with single-base-alterations, which makes them very useful as markers for identification of individual Beagles and useful for parentage testing. PMID- 11936867 TI - In vitro-prepared advanced glycation end-products and the modulating potential of their low-molecular weight degradation products in IRPTC-A rat proximal-tubular derived kidney epithelial cell line. AB - Low-molecular advanced glycation end-products (AGEs)-degradation products resulting from a proteolysis of tissue or circulating AGEs represent up to 80% of AGE plasma immunoreactivity. These AGE peptides contribute to the dramatic increase in AGE levels in end-stage renal disease even in the absence of diabetes. Because glomerular filtered AGE-degradation products may accumulate within intracellular compartments of proximal tubular epithelial cells, we investigated whether there is a pathway potentially mediating damaging effects of AGE-degradation products by perturbation of the function of the tubuloepithelium. Proximal tubular-derived rat kidney cells (IRPTC) were incubated with high molecular AGEs highly modified by incubation of bovine serum albumin (BSA) with glucose for 50 days in vitro, and with low-molecular AGE-degradation products derived from proteolytic cleavage and isolated in the molecular range between 1 and 30 kDa. The proliferation of IRPTC (3H-thymidine incorporation) was reduced to 89+/-1% and 69+/-2% after 24 hr of incubation with BSA-AGE and BSA-AGE degradation products, respectively. The cell viability of IRPTC was reduced significantly to 59+/-15% and 31+/-13% after 144 hr of incubation with BSA-AGE and BSA-AGE-degradation products, respectively. Conditioned media obtained from IRPTC incubated for 72 hr with BSA-AGE and its degradation products increased the proliferation rate of renal fibroblasts (RFb) to 222+/-24% and 449+/-40%, respectively. Incubation of IRPTC with BSA-AGE-degradation products increased the expression of endothelin-1 (ET-1) mRNA to 210% after 1 hr; the expression of platelet-derived growth factor-B (PDGF-B) mRNA reached 184% after 2 hr. Regarding the toxicity of AGEs to the kidney, low-molecular weight AGE-degradation products possibly form an individual fraction with a comparatively higher toxic potential. PMID- 11936868 TI - Molecular mechanism of TGF-beta1-induced apoptosis in HC11 mouse mammary epithelial cells (MEC). AB - The involvement of p53, Bax, cytochrome C and CPP-32 (caspase-3) in the molecular mechanism ofTGF-beta1-induced apoptosis in HC11 mouse mammary epithelial cells (MEC) was examined. Laser scanning cytometry (LSC) was applied for the quantitative analysis of expression and distribution of examined apoptosis related proteins in the cytoplasmic (Cf) and nuclear (Nf) area. Maximal pixel of fluorescence (MP) parameter corresponding to aggregation of molecules in the cell was also measured. Confocal and immunoelectron microscopy were used as a complementary methods. Apoptosis induced by TGF-beta1 (2 ng/ml) was associated with the increase of Bax MP observed within 60 min. after cytokine administration, indicating aggregation of Bax in the cell. Immunoelectron microscopy revealed Bax aggregation on mitochondrial membranes, rough endoplasmic reticulum, Golgi apparatus, cytoskeleton, nuclear envelope and inside of nucleus. The accumulation of Bax in the nucleus was confirmed by compartmental Bax analysis, showing the increase of cell number with elevated Bax Nf in 2 hr after TGF-beta1 administration to the culture. The redistribution of Bax within the cell was dependent on its activation occurring by the cleavage at N-terminal epitope and exposure of BH3 domain. Bax aggregation on organelles was completely abolished by prolactin or IGF-I. TGF-beta1 increased p53 MP, evidently after 4 hr of cell culture exposure to this cytokine. p53 was accumulated first of all in the nucleus, which was shown by significant increase of p53 Nf/Cf ratio and increase of p53-related nuclear fluorescence on confocal images. TGF-beta1 decreased cytochrome C MP, which corresponded to its release from mitochondria and dissipation in the cytosol. It was accompanied by the increase of CPP-32 MP and concentration of 89 kDa product of PARP degradation in the nucleus. In conclusion, TGF-beta1 triggers apoptosis in MEC through mitochondrial pathway involving: activation and translocation of Bax to mitochondrial membranes, release of cytochrome C from mitochondria, activation of CPP-32 and degradation of its substrate - PARP in the nucleus. Activation and subcellular redistribution of Bax is inhibited by lactogenic hormones: prolactin and IGF-I. PMID- 11936869 TI - Towards versatile metal associating substrates for the determination of peroxidatic activity/hydrogen peroxide by chemical designing of Schiff base derivatives. AB - Novel chromogenic N-arylmethyl-aniline-substrates of the general formula R-NH CH2C6H5-n-Xn (X = OH, NHR) for the localization of peroxidatic activity/hydrogen peroxide were synthesized in two steps from starting amines and aromatic aldehydes. When using 1,2-dinucleophiles (e.g. diaminobenzidine) as starting material there may be limitations resulting from dominant altemative reaction courses (amino-imines vs bis-imines) or tautomerism (amino-imines vs benzimidazolines). This has been investigated in a model study on 1,2 phenylendiamine. All substrates were evaluated for application in histochemistry, electrophoresis, colorimetry and electron microscopy. Thus, 1/ endogenous peroxidatic activity in native cryotome sections of Wistar rats was stained. One selected reagent was used for immuno-histochemical demonstration of vimentine and applied for laser microscopy at 543 nm as well. 2/ Electro-blotted dilution series of horseradish peroxidase were stained and reagents ranked according to their sensitivity. 3/ In test tube experiments precipitation behavior, color and solubility of precipitates was investigated. 4/The chromogens are capable of forming electron opaque final reaction products by way of increased osmiophilicity of the specific reaction product or/and by complexation of electron dense metals as demonstrated by electron microscopical investigations. As a result, two novel reagents derived from 1,2-phenylendiamine and 2 aminophenol are recommended especially for electron microscopy: The discrimination between internum and extemum of specific granules after osmium tetroxide treatment is resolved clearly as compared with results obtained with the standard Kamovsky protocol. PMID- 11936870 TI - Translational errors as an early event in prion conversion. AB - A prion is an infectious, altered form of a cellular protein which can self propagate and affect normal phenotype. Prion conversion has been observed for mammalian and yeast proteins but molecular mechanisms that trigger this process remain unclear. Up to now, only post-translational models have been explored. In this work, we tested the hypothesis that co-translational events may be implicated in the conformation changes of the Ure2p protein of Saccharomyces cerevisiae. This protein can adopt a prion conformation leading to an [URE3] phenotype which can be easily assessed and quantified. We analyzed the effect of two antibiotics, known to affect translation, on [URE3] conversion frequency. For cells treated with G418 we observed a parallel increase of translational errors rate and frequency of [URE3] conversion. By contrast, cycloheximide which was not found to affect translational fidelity, has no influence on the induction of [URE3] phenotype. These results raise the possibility that the mechanism of prion conversion might not only involve alternative structures of strictly identical molecules but also aberrant proteins resulting from translational errors. PMID- 11936871 TI - Functional characterization of the promoter region of the human mu opioid receptor (hMOR) gene: identification of activating and inhibitory regions. AB - The mu opioid receptor (MOR) is thought to mediate a variety of morphine's effects, including analgesia and addiction. The expression of opioid receptors can be up and down regulated, but little is known about molecular processes that regulate expression of the MOR gene. To study the regulatory elements that control expression of the human MOR (hMOR) gene, 2325 bp of the 5'-regulatory sequence of the hMOR gene were cloned and sequenced. A transcription initiation site (TIS) was mapped 252 (-252) nucleotides upstream from the translation start site (+1) by primer extension experiments using human thalamus poly(A)+ mRNA. In addition, several putative distal TISs were also identified; the most distal site was mapped 663 bp upstream of the translation start site. A series of 5'-deleted hMOR promoter-luciferase constructs were made and transiently transfected into a MOR expressing neuroblastoma cell line, SK-N-SH, and a non-expressing cell line, HeLa. These transient transfection studies indicated that the region from -563 to -292 contained a strong enhancer element(s), while the region from -776 to -564 possessed a repressor element(s). A similar transfection pattern was observed with SK-N-SH and HeLa cells, suggesting that there is not a tissue-specific element in the region from -2325 to -252. PMID- 11936872 TI - Characterization of HIV isolates from Puerto Rican maternal-infant pairs reveal predominance of non-syncytium inducing (NSI) variants with CCR5 genotype. AB - In this study, HIV-1 variants from a cohort of forty-eight Puerto Rican pregnant women and their 50 infants (one had triplets), were isolated and characterized, in order to determine the type of HIV-1 variants that are predominantly transmitted. All were enrolled in the prenatal AIDS Clinical Trials Group (ACTG) and received anti-retroviral therapy. Fifteen of the 50 infants (30%) were positive by V3 PCR suggesting that they harbored a copy of the HIV envelope gene. Three of 50 infants (6%) were HIV-1 culture and PCR positive, indicating active infection. HIV-positive cultures were obtained from 32 of the 48 mothers. Sixty two percent of the isolates (20/32) were macrophage-tropic and non-syncytium inducing, three percent (1/32) had dual tropism, and thirty four percent (11/32) were non-syncytium inducing and did not grow in macrophages. Phenotype and genotype of the HIV variants from the three infected infants revealed the presence of macrophage-tropic and non-syncytium-inducing strains. Genotype analysis of the HIV env V3 loop revealed the presence of specific amino acids that are predictive of CCR5 usage. Sequence analysis of the HIV pol gene from the three infected infants indicated that vertical transmission was not caused by the presence of antiviral resistance mutations. These results indicate that mothers undergoing antiretroviral treatment at different stages of the disease and with different viral loads transmit predominantly macrophage-tropic/non-syncytium inducing/CCR5 variants to their infants. PMID- 11936873 TI - On the structural form of iron in ferritin cores associated with progressive supranuclear palsy and Alzheimer's disease. AB - Recently, Quintana and others published the results of TEM investigations of ferritin cores extracted from the brain tissue of patients suffering from progressive supranuclear palsy (PSP) and Alzheimer's disease (AD). These ferrihydrite (fFe2O3.9H2O) cores from the iron storage protein ferritin were found to contain a ferrous (Fe2+) iron-bearing biomineral with cubic structure similar to the magnetite standards which were measured. This is a highly important result as magnetic and electron microscopy analysis has demonstrated the presence of magnetite (Fe3O4) and maghemite (gammaFe2O3) in human brain tissue. PMID- 11936874 TI - Is decreased HIV-1 infectivity of placental macrophages caused by high levels of beta-chemokines? AB - The objective of this study was to determine the effect of beta-chemokine secretion on HIV infection of placental macrophages (HF) as compared to monocyte derived macrophages (MDM). For this purpose, we measured chemokine production in supematants of LPS stimulated and unstimulated HF and MDM. LPS stimulated cultures produced 3 to 10 times higher levels of MIP-1alpha, MIP-1beta and RANTES than unstimulated cultures. The level of MIP-1beta was the highest of the three chemokines secreted upon stimulation of HF cells. Cell cultures were inoculated with HIV-BaL, a R5 virus, and tested for p24 antigen and chemokine production at days 5 and 10 post-infection (P.I.). We did not find significant differences in the level of chemokines produced by HIV-1-infected and uninfected MDM and HF cells. However, significant differences were found in p24 antigen released by unstimulated and LPS stimulated cells. In contrast to HF cells, MDM cultures showed a significant inhibition of p24 antigen production when cells were stimulated with LPS prior to infection. HF cells were less susceptible to HIV-1 infection than MDM, and chemokines produced by HF cells did not result in further inhibition of HIV-1 infection. We found that in contrast to MDM, decreased susceptibility HF cells to HIV infection is not due to increased levels of chemokines, but to decreased HIV-1 coreceptor expression. PMID- 11936875 TI - Lactacystin enhances cisplatin sensitivity in resistant human ovarian cancer cell lines via inhibition of DNA repair and ERCC-1 expression. AB - Cisplatin is among the most effective chemotherapeutic agents in the treatment of human ovarian cancer. The cytotoxicity of cisplatin results primarily from its ability to bind covalently to DNA and prevent DNA replication and transcription. The ubiquitin-proteasome pathway plays important roles in a broad array of basic cellular processes. Lactacystin is a selective inhibitor of the proteasome that can inhibit the ubiquitin pathway. However, the effect of lactacystin on DNA repair and the antitumor activity of cisplatin in ovarian cancer have not been evaluated. We report in this work that lactacystin, at concentrations that do not appear harmful, increased cisplatin toxicity in three resistant human ovarian carcinoma cell lines. In addition, lactacystin significantly enhanced DNA platination and decreased DNA repair of cisplatin-DNA adducts in these cell lines, as measured by atomic absorption spectrometry. Furthermore, Northem blot analysis and in vitro nuclear transcript elongation assay demonstrated that lactacystin dramatically reduced the steady-state mRNA expression and the rate of transcription of the DNA repair gene ERCC-1 in these cells. These observations indicate that proteasome inhibition has impact on nucleotide excision repair in several ways: i/ the normal ERCC-1 message upregulation is suppressed; ii/ cisplatin-DNA adduct repair is inhibited, and iii/ DNA platination, as well as cisplatin cytotoxicity, is enhanced. PMID- 11936876 TI - The infrared spectroscopic Gram stain. AB - The attenuated total reflectance (ATR) spectra of bacterial colonies were obtained in situ, without their removal from the agar growth media using the infrared microscope. Principal components regression (PCR) analysis was used to obtain a measure of the differentiating potential of the ATR spectra obtained on two standard sets consisting of 31 and 44 species, respectively, grown on trypticase soy agar. A quantitative response value of +1 was assigned to Gram positive species and -1 was assigned to Gram negative species. In a cross validation experiment, the Gram stain property correlated with the factor loadings with a multiple correlation of 0.9502 and 0.9520 for each of the standard sets, respectively. Also, it was found that ATR spectra can be obtained from bacteria grown on blood agar and still have good differentiating capabilities for Gram stain. In addition, ATR spectra provide significantly better differentiating spectral information and predicting capabilities than specular reflectance spectra obtained in situ. Finally, the numerical value obtained for the Gram stain can be used as a measure of Gram variability. Results indicate that the Gram positive values shift to Gram negative values as the bacterial cell wall deteriorates with time. PMID- 11936877 TI - The angiogenic factor heparin affin regulatory peptide (HARP) induces proliferation of human peripheral blood mononuclear cells. AB - Heparin affin regulatory peptide (HARP) also named pleiotrophin (PTN) is a polypeptide that belongs to a family of heparin-binding molecules. HARP displays mitogenic activity for a wide variety of cells, including fibroblast, endothelial and epithelial cells. This study reports, to our knowledge for the first time that HARP induced the stimulation of triated thymidine incorporation in quiescent human peripheral blood mononuclear cells in a dose-dependant manner, measured after 7 days of culture. Maximal stimulation was observed at picomolar concentration with ED50 corresponding to the half maximum effect at 50 pM. In contrast, midkine (MK), a related heparin-binding growth/differentiation factor, with more than 50% amino acid sequence homology with HARP was ineffective. These results suggest that HARP could be considered as a new cytokine involved inthe growth regulation of cell mediated immunity. PMID- 11936878 TI - Characterization of first trimester human fetal placental vessels using immunocytochemical markers. AB - Cell differentiation markers on placental villi from the first trimester of human pregnancy have been studied by indirect immunofluorescence. Fluorescence labelling with antibodies against CD34 and CD31 was conspicuous in the vascular cells. The vascular paracellular clefts were labelled by anti-cadherin-5. A few vascular cells exhibited a positive reaction for von Willebrand factor, high molecular-weight melanoma-associated-antibody and alpha-sm-actin compared to term pregnancy, indicating changes in protein expression during vascular differentiation. The poor anti-collagen IV reaction and the absence of a sm myosin fluorescent signal observed around the vessels confirned the immaturity of the vessels. In contrast, strong reactions have previously been obtained with the latter antibodies in similar locations using term placental villi. A labelling was observed for antibodies against alpha3 and alpha5 integrins in these immature placental vessels suggesting cell-matrix interactions with specific domains of laminin or fibronectin. The vascular cells were also stained by anti-CD26. Surprisingly, the fetal vascular cells exhibited immunostainings in common with the villous cytotrophoblast (CD26) or the syncytiotrophoblast (cadherin-5) and cell islands cytotrophoblast (CD31, cadherin-5, alpha3 and alpha5 integrin subunits). These observations suggested a two step process for fetal vasculogenesis in the villi: i/ the formation of peripheral vessels induced by growth factors or cytokines derived from the nearby trophoblast, ii/ the development of muscular vessels due to growth factors or cytokines production induced by circulatory changes. PMID- 11936879 TI - Actin in semithin sections of myocardial biopsies as a tool to visualize myofibrillary degradation in humans. AB - Dilated cardiomyopathy (DCM) is characterized by dilation of ventricular walls and the reduction of cardiac contractillity, caused by loss of myofibrils and apoptosis of the myocytes. Histochemical assessment of myocardial biopsies is commonly used to monitor these events. Here we report the use of immunohistochemistry for alpha-actin in semithin serial sections of myocardial biopsies taken from patients with various stages of DCM. This technique allows also the detection of DNA-fragmentation and tissue transglutaminase as markers for cell damage in subsequent sections of the same myocytes. Intense actin immunostaining was observed only in the cytoplasm of cells that also expressed tissue transglutaminase in the cytoplasm and showed DNA-fragmentation in the nucleus. We assume that endogenous proteolysis, associated with apoptotic events enhances the stainability of actin in semithin sections. High resolution detection of actin probably visualizes myofibrillary deterioration, which may correspond to reduced cardiac performance in a more direct manner than the histochemistry of apoptotic markers. PMID- 11936881 TI - A veterinary pot-pourri at the AVS congress in Edinburgh. Association of Veterinary Students. PMID- 11936880 TI - Optical detection of mitochondrial NADH content in intact human myotubes. AB - Time-gated fluorescence spectroscopy in combination with non-radiative energy transfer was used on intact human skeletal myotubes for the determination of the mitochondrial NADH content which is considered to be a sensitive indicator of mitochondrial function. To mimic dysfunction of the mitochondrial energy metabolism, complexes I or III of the respiratory chain were inhibited by drugs. In the absence of the fluorescent mitochondrial marker rhodamine (R123), the NADH autofluorescence (i.e. a signal monitoring cytoplasmic plus mitochondrial NADH) remained unchanged upon inhibition of complex I by rotenone, and was increased by a factor of 2 upon inhibition of complex III by antimycin. In the presence of R123, the autofluorescence of NADH was reduced indicating non-radiative energy transfer from NADH to R123. The ratio of the R123 fluorescence signals obtained with the two excitation wavelengths of 355 nm and 488 nm was taken as a measure of mitochondrial NADH. Relative NADH changes were estimated in the presence of the above-mentioned inhibitors. Upon complex I inhibition, mitochondrial NADH was increased by a factor of 1.5. Upon inhibition of complex III, mitochondrial NADH was increased by a factor of 2. We conclude that time-gated spectroscopy combined with non-radiative energy transfer is an appropriate tool for probing mitochondrial enzyme complex deficiencies. PMID- 11936882 TI - Primary thoracic wall tumours of mesenchymal origin in dogs: a retrospective study of 46 cases. AB - Forty-three of the chest wall tumours in 46 dogs were malignant; five had metastases apparent at the time of presentation, five more had metastases discovered intraoperatively. Surgical resection of the tumours was associated with a significantly better outcome than conservative management. The median survival times after surgery for dogs with osteosarcoma was 17 weeks, for dogs with fibrosarcoma it was 26 weeks and for dogs with chondrosarcoma it was 250 weeks. En bloc excision of primary tumours affecting the chest wall was associated with minimal morbidity, but long-term survival was limited by distant metastases, primarily to the lungs. The tumours recurred in only three dogs. Early, radical surgical excision is recommended in the management of tumours of the chest wall. The prognosis depends on the histologic type of tumour and a histological diagnosis is mandatory before excision. PMID- 11936883 TI - Efficacy of avian pneumovirus vaccines against an avian pneumovirus/Escherichia coli O2:K1 dual infection in turkeys. AB - The clinical, pathological and microbiological outcome of a challenge with avian pneumovirus (APV) and Escherichia coli O2:K1 was evaluated in turkeys vaccinated with an attenuated APV vaccine and with or without maternally derived antibodies. Two groups of two-week-old poults, one with and one without maternally derived antibodies against APV, were vaccinated oculonasally with attenuated APV subtype A or B. A third group remained unvaccinated. Eleven weeks later, the turkeys were inoculated intranasally with either virulent APV subtype A, or E. coli O2:K1, or with both agents three days apart. After the dual infection, birds vaccinated with attenuated subtype A or B, and with or without maternally derived antibodies, had lower mean clinical scores than the unvaccinated birds. In the vaccinated birds, virus replication was significantly reduced and no bacteria were isolated, except from the birds vaccinated with attenuated subtype B. In the unvaccinated turkeys, large numbers of E. coli O2:K1 were isolated from the turbinates of the dually infected birds between one-and-a-half and seven days after they were inoculated. PMID- 11936884 TI - Pharmacokinetics of ivermectin in llamas (Lama glama). AB - The pharmacokinetic behaviour of ivermectin was investigated in adult llamas (Lama glama) by using high performance liquid chromatography with a lower limit of quantification of 2 ng/ml to measure its concentration in serum. Llamas were treated with one of three commercial formulations (injectable, pour-on or oral paste) at dosages recommended by the manufacturer, or with an experimental injectable sustained-release formulation. In five llamas given 1 per cent ivermectin subcutaneously at 200 microg/kg, the median peak serum concentration (Cmax) was 3 ng/ml and the area under the serum concentration-time curve (AUC) was 13.5 ng x day/ml. In six llamas treated topically with 0.5 per cent ivermedin pour-on at 500 microg/kg, Cmax was 2.5 ng/ml or less and the AUC was 7.75 ng x day/ml or less. In seven llamas with measurable concentrations of ivermedin, the median times to peak serum concentration (tmax) were six days after subcutaneous injection and seven days after treatment with the pour-on formulation. In six llamas, the serum concentration of ivermectin remained less than 2 ng/ml for 124 hours after treatment with a 1.87 per cent oral paste at 200 microg/kg. In five llamas treated subcutaneously with 25 per cent ivermectin sustained-release microspheres at 1500 microg/kg, the median Cmax was 5 ng/ml and the median AUC was 224 ng x day/ml. PMID- 11936885 TI - Trends in prevalence of BSE in Switzerland based on fallen stock and slaughter surveillance. PMID- 11936886 TI - Testicular teratoma in a three-day-old thoroughbred foal. PMID- 11936887 TI - Administration of enrofloxacin and capsaicin to chickens to achieve higher maximal serum concentrations. PMID- 11936888 TI - Urachal diverticulum in a kid. PMID- 11936889 TI - Diphtheria toxin production by Corynebacterium ulcerans from cats. PMID- 11936890 TI - Future of the RCVS library. PMID- 11936891 TI - Future of the RCVS library. PMID- 11936892 TI - Cattle transport in the UK. PMID- 11936893 TI - Helminth control in sheep. PMID- 11936895 TI - A 13-week feeding study in the rat with shea oleine and hardened shea oleine. AB - Shea oleine is an oil fraction derived from the nut of the tree, Butyrospermum parkii, which grows in central and western Africa. There are several uses of shea oleine, including its use as a cooking oil and, after hardening, in margarine and toffee fat. As part of a series of studies, a 13-week rat feeding study was conducted in which groups of Colworth-Wistar rats (15 male and 15 female) were fed 27.5% total fat semipurified diets containing 20% (w/w) shea oleine (SU) or hardened shea oleine (SH). Equivalent groups of rats were fed either 20% (w/w) palm oil (PU), soyabean oil (BU), or the hardened (hydrogenated) equivalents (PH and BH, respectively). Assessments of body weight, food and water intake, clinical pathology, organ weights, and macroscopic histopathology were carried out. Results showed that shea oleine diets produced similar biological effects to palm oil and and soyabean oil diets. A slightly reduced body weight gain was noted in rats fed either of the shea oleine diets in comparison with palm oil and soyabean oil. The process of hardening had no significant impact on the normal growth in rats fed shea oleine, although minor differences compared with unhardened diets comprised some small changes in clinical chemistry parameters, raised liver weight, and less liver lipid. In addition, raised alkaline phosphatase and an increase in food intake were noted in rats fed SH. All diets were well tolerated in the growing rat and none of the findings in the study were considered to be adverse. PMID- 11936897 TI - ACT Lifetime Contribution Award, 2001. Dr. Arthur Furst--awardee. American College of Toxicology. PMID- 11936896 TI - Toxicological evaluation of 1,2-bis(methylsulfonyl)-1-(2-chloroethyl)-2 (methylaminocarbonyl)hydrazine (VNP40101M), a novel alkylating agent with potential antitumor activity, with intravenous administration in rats and dogs. AB - These studies investigated the toxicological effects of 1,2-bis(methylsulfonyl)-1 (2-chloroethyl)-2-(methylaminocarbonyl) hydrazine, VNP40101M, a novel alkylating antitumor agent, in animals. Sprague-Dawley rats (2-10/sex/time point at each dose) and Beagle dogs (1-3/sex/time point at each dose) were treated with VNP40101M (0 [vehicle], 1, 3, 10, and 20 mg/kg in rats and 0, 0.3, 1, and 3 mg/kg in dogs), given intravenously (IV, bolus via the tail or slow push via the cephalic or saphenous vein, respectively) once daily for 5 consecutive days. Clinical signs, mortality, body weight, clinical pathology, gross necropsy, organ weights, and histopathology were evaluated for as long as 43 days in rats and 50 days in dogs. In rats, the toxic doses were found to be at 10 and 20 mg/kg, which induced mainly pulmonary toxicity and mortality. The pulmonary toxicity was reflected by an increase in lung weight; clear, pink or red fluid within the thoracic cavity observed at necropsy; and histopathological evidence of alveolar edema, vascular congestion, alveolar histiocytosis, and vascular thrombi. Although some of these effects were observed in rats treated with 3 mg/kg, the incidence was low (approximately 7%-30%) and may be reversible (based on the time dependent reduction in the magnitude of lung weight increases). Therefore, the maximum tolerated dose (MTD, or the maximum dose that did not induce significant toxicity or induced reversible toxicity) was > or = 3 mg/kg. VNP40101M at 1 mg/kg did not induce any toxicity, other than low incidence of alveolar edema (2/30 rats), and increased incidences of capillary ectasis/congestion and alveolar histocytosis (2-6/30 rats vs. 1/30-36 in control rats). Therefore, the low effect level (LOEL) is considered to be 1 mg/kg in rats when given IV for 5 days. In dogs, LOEL, MTD, and toxic dose levels were comparable (based on a body weight/surface area conversion) to those in rats, except for some gastrointestinal (GI) effects (i.e., red lesion in the ileum) observed at 0.3 mg/kg (equivalent to 1 mg/kg, or similar to the LOEL in rats) and the associated effects (slight body weight loss and inappetence). For dogs treated with 1 mg/kg (equivalent to approximately 3 mg/kg, or MTD, in rats), VNP40101M induced the same GI effects seen in dogs treated with 0.3 mg/kg of VNP40101M. Additionally, a transient reduction in white blood cell counts was also observed. Three mg/kg (equivalent to approximately 10 mg/kg, or toxic dose level, in rats) was toxic to dogs, as reflected by the poor clinical condition of these dogs, which subsequently required euthanasia. In conclusion, VNP40101M, when given IV once daily for 5 consecutive days, has a LOEL of 1 mg/kg, a MTD of 3 mg/kg, and toxic doses at > or = 10 mg/kg in rats. The primary toxicity of VNP40101M was pulmonary toxicity and mortality. Based on an interspecies body weight/surface area conversion, VNP40101M had comparable LOEL (0.3 mg/kg), MTD (1 mg/kg), and toxic doses (> or = 3 mg/kg) in dogs, except that dogs appeared to be more sensitive to the GI effects of VNP40101M. PMID- 11936898 TI - Inhalation toxicology of octamethylcyclotetrasiloxane (D4) following a 3-month nose-only exposure in Fischer 344 rats. AB - Octamethylcyclotetrasiloxane (D4) is a low-molecular-weight cyclic siloxane used primarily in the synthesis of silicone polymers. The objective of the present study was to evaluate the subchronic toxicity of D4 following a 3-month nose-only inhalation exposure. Male and female Fischer 344 rats (20/sex/group) were exposed 6 h/day, 5 days/week for 3 months to vapor concentrations of 0, 35, 122, 488, and 898 ppm D4. Also, an additional 10 per sex in the control and high-exposure groups were allowed a 4-week recovery period to observe reversibility, persistence, or delayed occurrence of any potential adverse effects. Body weights and food consumption were monitored at least twice weekly over the course of exposures. Approximately 18 hours preceding euthanasia, animals were transferred into metabolism cages for urine collection, and were fasted. At necropsy, rats were anesthetized with pentobarbital and euthanized by exsanguination. Blood was collected for hematological and clinical biochemical analyses. Selected organ weights were measured and a complete set of tissues was taken for histopathological examination. A concentration-dependent increase in absolute and relative liver weight (488 to 898 ppm) and a significant decrease in ovarian weight (898 ppm) were observed in female rats. Exposure to D4 via nose-only inhalation (35 to 898 ppm) produced minor alterations in hematological and serum chemistry parameters that were considered either incidental and of little toxicological significance (hematology) or suggestive of metabolic adaptation/alteration (serum chemistry) in response to exposure-related hepatomegaly. There were no histopathological findings noted in the liver. Histopathological evidence indicated the primary target organs following D4 inhalation exposure to be components of the female reproductive tract. Reversible histopathological changes were observed in the ovary (hypoactivity) and vagina (mucification) of female rats in the high-dose group only (898 ppm). Although an increase in the incidence and severity of both macrophage accumulation, interstitial inflammation, and eosinophil infiltration was observed in the lungs of male and female rats exposed to D4, the toxicological significance is uncertain as other inhalation studies at similar concentrations failed to show these effects. In summary, nose-only inhalation of a high concentration of D4 resulted in reversible histopathological changes in the female rat reproductive tract. Lower concentrations did not elicit these same effects. PMID- 11936899 TI - Distinguished Service Award. Dr. Curtis D. Klaassen--awardee. PMID- 11936900 TI - Use of genetically engineered mice in drug discovery and development: wielding Occam's razor to prune the product portfolio. AB - Genetically engineered mice (GEMs) that either overexpress (transgenic) or lack (gene-targeted, or "knock-out") genes are used increasingly in industry to investigate molecular mechanisms of disease, to evaluate innovative therapeutic targets, and to screen agents for efficacy and/or toxicity. High throughput GEM construction in drug discovery and development (DDD) serves two main purposes: to test whether a given gene participates in a disease condition, or to determine the function(s) of a protein that is encoded by an expressed sequence tag (EST, an mRNA fragment for a previously uncharacterized protein). In some instances, phenotypes induced by such novel GEMs also may yield clues regarding potential target organs and toxic effects of potential therapeutic molecules. The battery of tests used in phenotypic analysis of GEMs varies between companies, but the goal is to define one or more easily measured endpoints that can be used to monitor the disease course--especially during in vivo treatment with novel drug candidates. In many DDD projects, overt phenotypes are subtle or absent even in GEMs in which high-level expression or total ablation of an engineered gene can be confirmed. This outcome presents a major quandary for biotechnology and pharmaceutical firms: given the significant expense and labor required to generate GEMs, what should be done with "negative" constructs? The 14th century philosophical principle known as Occam's razor-that the simplest explanation for a phenomenon is likely the truth-provides a reasonable basis for pruning potential therapeutic molecules and targets. In the context of DDD, Occam's razor may be construed to mean that correctly engineered GEMs lacking obvious functional or structural phenotypes have none because the affected gene is not uniquely essential to normal homeostasis or disease progression. Thus, a "negative" GEM construct suggests that the gene under investigation encodes a ligand or target molecule without significant therapeutic potential. This interpretation indicates that, at least in a market-driven industrial setting, such "negative" projects should be pruned aggressively so that resources may be redirected to more promising DDD ventures. PMID- 11936901 TI - Evaluation of the Tg.AC transgenic mouse assay for testing the human carcinogenic potential of pharmaceuticals--practical pointers, mechanistic clues, and new questions. AB - Transgenic mouse strains with genetic alterations known to play a role in the multistage process of carcinogenesis are being used increasingly as models for evaluating the human carcinogenic potential of chemicals and pharmaceuticals. The Tg.AC transgenic mouse is one of the strains currently being used in such alternative short-term carcinogenicity testing protocols. This review is focused on recent data from studies designed to evaluate this model's ability to discriminate carcinogens from noncarcinogens. Details relating to protocol design that can significantly impact study outcome are described. Data relating to mechanisms of chemical tumor induction in the Tg.AC model are reviewed, and questions have been formulated to encourage research to further guide appropriate future applications of this model. PMID- 11936902 TI - Xenobiotic transporters: another protective mechanism for chemicals. AB - Xenobiotic transporters are responsible for the uptake of some chemicals into cells, and extremely important for the export of chemicals out of cells. A number of families of xenobiotic transporters have been cloned the last few years. Some microsomal enzyme inducers will enhance the plasma disappearance and biliary excretion of some xenobiotics that are not biotransformed in the intact animal, as well as in isolated hepatocytes. This is due to an up-regulation of xenobiotic transporters. As a result, some microsomal enzyme inducers will enhance the elimination and decrease the toxicity of some chemicals by enhanced transport. PMID- 11936903 TI - Experimental analysis of behavior of homing pigeons as a result of functional disorders of their lagena. AB - Behavioral experiments concerning the homing abilities of pigeons were done by sectioning their lagenal nerves or interfering with the function of the lagena using a magnet. Twenty-one birds were treated in this way and 30 birds from the same loft of racing pigeons were used as controls. The results of homing tests clearly revealed a magnetic influence on the function of the lagena in terms of the navigation ability of pigeons: the treated birds were either lost or significantly delayed while the controls returned within 30 min of release. The lagena of birds is a unique organ and it is concluded that it is a key element in the magnetic sensory system of birds. PMID- 11936904 TI - Change of guinea pig inner ear pressure by square wave middle ear cavity pressure variation. AB - The inner ear fluid pressure of guinea pigs was measured during square wave middle ear cavity pressure variation. Time constants were derived for the slopes of the inner ear pressure recovery curves after middle ear pressure change. A "single exponential" function did not fit well and therefore more complicated functions were used for this purpose. For middle ear pressure increasing from zero to a few centimetres of water, returning to zero again, decreasing from zero to minus a few centimetres of water and then returning to zero again, time constants for the inner ear pressure recovery curves were on average 15.0, 8.6, 2.5 and 2.5 s, respectively. The results could not be described using a linear model with constant window membrane compliance and cochlear aqueduct flow resistance. A possible explanation for the large difference in time constants for positive or negative middle ear pressure changes is a dependence on aqueduct flow resistance or round window membrane position. PMID- 11936905 TI - In vitro study of IL-8 and goblet cells: possible role of IL-8 in the aetiology of otitis media with effusion. AB - One of the main characteristics of otitis media with effusion (OME) is the differentiation of basal cells into goblet cells with subsequent proliferation in a modified respiratory epithelium leading to the formation of mucin-rich effusion in the middle ear cleft. In order to determine the effect of pro-inflammatory cytokines identified in OME, e.g. IL-1beta, tumour necrosis factor (TNF)-alpha, IL-6 and IL-8, on goblet cells, and to clarify the role of IL-8 in particular, we used the human goblet cell line HT29-MTX, which secretes two OME-related mucins: MUC5AC and MUC5B. IL-1beta and TNF-alpha stimulated the secretion of IL-8 in HT29 MTX goblet cells. Dose- (2-200 ng/ml) and time- (0-5 days) response studies of IL 8-induced mucin secretion were carried out. IL-8 upregulated the secretion of MUC5AC and MUC5B mucins in a concentration-dependent manner, with a maximum response at an IL-8 concentration of 20 ng/ml. IL-8 (20 ng/ml)-mediated mucin secretion persisted for up to 5 days, with a peak response 72 h after the addition of cytokine. These results suggest that: (i) goblet cells are target cells for the pro-inflammatory cytokines IL-1beta, TNF-alpha and IL-8 and can contribute to the pathogenesis of OME by increasing both the concentration of IL 8 and the secretion of mucin; and (ii) IL-8 stimulates prolonged mucin secretion from goblet cells and may be involved in the maintenance of the disease in the chronic stage. PMID- 11936906 TI - Induction of mucous cell metaplasia in the middle ear of rats using a three-step method: an improved model for otitis media with mucoid effusion. AB - Otitis media with mucoid effusion, characterized by mucous cell metaplasia in the middle ear cleft and thick fluid accumulation in the middle ear cavity, is a common otological disease that frequently affects young children. Multiple factors are involved in the development of this disease, especially middle ear infection and Eustachian tube dysfunction. In this study, in order to induce otitis media with effusion in rats, we introduced a three-step method, namely inoculation of Streptococcus pneumoniae at 10(7) colony-forming units (CFU)/ear or Haemophilus influenzae at 5 x 10(7) CFU/ear into the middle ear cavity twice at 2-week intervals, followed by Eustachian tube obstruction (ETO) for 4 and 8 weeks. Animals inoculated with phosphate-buffered saline (PBS) twice in the same manner followed by ETO served as controls. Middle ear effusion and mucosa were harvested for evaluation of carbohydrate concentrations and mucous cell density, respectively. We found that rats inoculated with S. pneumoniae twice, followed by ETO at 8 weeks, yielded the highest carbohydrate concentration in middle ear effusion and the highest goblet cell density in the middle ear cavity compared to the H. influenzae and PBS groups. It is tentatively concluded that inoculation of S. pneumoniae at 10(7) CFU/ear into the middle ear cavity of rats twice at 2-week intervals, followed by ETO for 8 weeks, is a promising animal model for otitis media with mucoid effusion which may be valuable for studying the human counterpart. PMID- 11936907 TI - Expression of activating transcription factor 3 and growth-associated protein 43 in the rat geniculate ganglion neurons after chorda tympani injury. AB - The purpose of this study was to evaluate the degree of damage in the geniculate ganglion and its target organ as a result of chorda tympani (CT) injury. We performed unilateral transection of the rat CT and examined expression of the activating transcription factor 3 (ATF3), a neuronal injury marker, and the growth-associated protein 43 (GAP-43), a regeneration-associated molecule. The mean proportion of ATF3-immunoreactive (ir) neurons in the geniculate ganglion was approximately 32% at 3 days after CT injury, but these neurons were never detected in the naive ganglion. Using in situ hybridization, the mean percentage of GAP-43 mRNA-labeled neurons (signal : noise ratio > or = 10) was observed to have increased significantly to approximately 60% for 1-7 days after CT injury, while that in the naive ganglion was < 15%. The results of morphological studies using scanning electron microscopy and immunohistochemistry indicated that atrophic change and reduction of protein gene-product 9.5-ir fibers in the denervated papillae, mainly in the intragemmal region, were observed after CT injury. Increase in GAP-43 mRNA, suggesting CT axonal regeneration, may have a role in recovery from taste disorders. However, this regenerative process may be involved in abnormal activity in the axotomized neurons or the adjacent intact neurons and so one must not disregard the existence of injured geniculate ganglions when considering the treatment of diseases that cause CT injury. PMID- 11936908 TI - Myringotomized tympanic membranes cultured in vitro do not develop myringosclerosis. AB - The aim of this study was to evaluate the development of myringosclerosis in in vitro-cultured tympanic membranes. Sprague-Dawley rats were myringotomized bilaterally and the tympanic membranes were excised after sacrifice. The explants were placed in inserts in wells filled with a nutrient medium. Every second day the tympanic membranes were photodocumented and after 9 days the explants were prepared for histology. On the 4th day the explants had attached to the bottom of the inserts and the specimens had thickened. From the perforation borders and the dissection edges a thin outgrowth was extending across the surface. By Day 9 the perforation had clearly diminished in size when examined in a stereomicroscope. In a light microscope the keratin layer was seen to protrude towards the centre of the perforation and, at the borders, epithelial cells were bridging the gap. Neither the pars tensa nor the pars flaccida showed any sclerotic lesions. The pars flaccida had thickened and the basal cells of the outer keratinized epithelium had invaded the connective tissue. Inflammatory cells were sparse in both the pars tensa and pars flaccida. The in vitro-cultured myringotomized tympanic membrane therefore shows a similar healing pattern to that in vivo. However, inflammatory reactions are sparse and there is no development of myringosclerosis. PMID- 11936909 TI - Use of electrocochleography for assessing endolymphatic hydrops in patients with Lyme disease and Meniere's disease. AB - From an otological standpoint, Lyme disease can manifest itself as Meniere's disease both clinically and electrophysiologically. The aim of this study was to describe the findings of routine clinical, auditory and vestibular tests in patients with Meniere's and Lyme disease and to use electrocochleography (ECoG) to assess the presence of endolymphatic hydrops (EH) in both diseases. Transtympanic ECoG was performed in 91 patients with Meniere's disease and in 11 patients with confirmed Lyme disease. In both diseases the majority of patients had more than one complaint. There was one case with isolated hearing loss in the Lyme disease group. Typical clinical manifestations of Meniere's disease (vertigo, sensorineural hearing loss and tinnitus) were found in 6/11 patients (54.5%) in the Lyme disease group. The ECoG results indicated that there were 65/91 patients (71.4%) with Meniere's disease and 5 patients (45.5%) with Lyme disease who presented with EH. No statistically significant difference was found between the incidence of different symptoms of Meniere's and Lyme disease. On the basis of these results, patients with Lyme disease should undergo careful examination and investigation, especially in endemic regions. The presence of EH does not exclude the presence of infection with borreliosis as a cause of Meniere's disease-like symptoms. PMID- 11936910 TI - The estimated incidence of symptomatic nasal polyps. AB - The objective of the present study was to establish the estimated incidence of symptomatic nasal polyps within a defined geographic area. Over a 6-year period all polyp patients diagnosed for the first time (n = 252; 174 males, 78 females) were prospectively registered at the study clinic. An estimated background population from which the patients were recruited was defined. Polyps were diagnosed by endoscopic examination. The mean estimated incidences for all age groups were 0.86 and 0.39 patients per thousand per year for males and females, respectively. The incidence increased with age, reaching peaks of 1.68 and 0.82 patients per thousand per year for males and females, respectively in the age group 50-59 years. The overall estimated incidence of symptomatic nasal polyps was 0.627 patients per thousand per year. Five percent of the patients had antrochoanal polyps and these patients had a lower mean age at diagnosis than the other patients, of whom 41% also had unilateral presentation at the time of diagnosis. This study seems to give a fairly reliable estimated incidence of symptomatic nasal polyps for different age groups and provides results that do not contradict prevalence studies in the literature, making it useful in a clinical context. PMID- 11936911 TI - An explanation for the seasonality of acute upper respiratory tract viral infections. AB - Despite a great increase in our understanding of the molecular biology of the viruses associated with acute upper respiratory tract viral infections (URTIs) there is a remarkable lack of knowledge and ideas about why URTI should exhibit a seasonal incidence. Most publications in this area either acknowledge a complete lack of any explanation for the seasonality of URTI or put forward an explanation relating to an increased "crowding" of susceptible persons in winter. This review will discuss some of the ideas concerning the seasonality of URTI and put forward a new hypothesis for discussion, namely that seasonal exposure to cold air causes an increase in the incidence of URTI due to cooling of the nasal airway. The hypothesis is supported by literature reports demonstrating that inhalation of cold air causes cooling of the nasal epithelium, and that this reduction in nasal temperature is sufficient to inhibit respiratory defences against infection such as mucociliary clearance and the phagocytic activity of leukocytes. A case is also made to suggest that warming of the nasal airway during fever and nasal congestion may help to resolve a current URTI. PMID- 11936912 TI - Symptoms and clinical and radiological signs predicting the presence of pathogenic bacteria in acute rhinosinusitis. AB - A minority of patients with upper respiratory tract infections (URTI) have a bacterial infection and may benefit from antibiotherapy. In previous investigations we showed that in patients suffering from acute rhinosinusitis associated with the presence of Streptococcus pneumoniae, Haemophilus influenzae or Moraxella catarrhalis in their nasopharygeal secretions, resolution of symptoms was significantly improved by antibiotic treatment. The present analysis was performed to determine whether specific clinical symptoms or signs observed during careful endoscopic examination of the nasal cavities could help the clinician to identify a subset of patients with moderate forms of acute rhinosinusitis infected with pathogenic bacteria. Detailed clinical histories were obtained and medical examinations performed in 265 patients (138 females, 127 males; mean age 35 years) presenting with a < 4-week history of URTI symptoms but who did not require immediate antibiotic therapy for severe rhinosinusitis. The presence of three pathogenic bacteria (S. pneumoniae, H. influenzae and M. catarrhalis) was determined in all patients by culture of nasopharyngeal secretions. Azithromycin (500 mg/day for 3 days; n = 133) or placebo (n = 132) were randomly given to all patients in a double-blind manner. Pathogenic bacteria were found in 77 patients (29%). The clinical signs and symptoms significantly associated in a multivariate model with the presence of bacteria included colored nasal discharge (p < 0.003), facial pain (p < 0.032) and radiologically determined maxillary sinusitis (complete opacity, air-fluid level or mucosal thickening > 10 mm) (p < 0.001). This best predictive model had a sensitivity of 69% and a specificity of 64% and therefore could not be used either as a screening tool or as a diagnostic criterion for bacterial rhinosinusitis. In the group of patients with positive bacterial cultures, resolution of symptoms at Day 7 was observed in 73% of patients treated with azithromycin and in 47% of patients in the placebo group (p < 0.007). We conclude that signs and symptoms of acute rhinosinusitis in patients with mild-to-moderate clinical presentations are poor predictors of the presence of bacteria. PMID- 11936913 TI - Study of ethmoidal mucosa in a population at occupational high risk of sinonasal adenocarcinoma. AB - The occurrence of nasal adenocarcinomas was first described in 1965. Since 1988 these tumors have been accepted as an occupational disease for woodworkers in Italy. There are several underlying reasons why there is interest in studying the ways in which sinonasal adenocarcinomas occur. Often diagnosed at advanced stages of development because their symptoms are non-specific, these tumors are associated with a high mortality rate. A multidisciplinary study protocol was developed in this investigation. The aim was to identify the factors and conditions that promote sinonasal tumor growth in a population at risk due to occupational exposure to wood dust. Sixty-eight carpenters with a minimum of 10 years exposure to wood dust were studied. The control group comprised 81 volunteers. The patients underwent the following protocol: completion of a case report form, physical examination, evaluation of nasal cavity patency, clinical laboratory tests and histological study of the nasal mucosa. Our study provides significant evidence of the elevated incidence of pavimentous metaplasia in workers occupationally exposed to wood dust. In addition, it underscores a significant deficit of immunoglobulin A in such workers compared to the controls. However, we did not find, as reported elsewhere in the literature, a statistically significant difference between cases and controls as regards nasal symptoms and hyperemia of the nasal mucosa. Our study showed that, even in the absence of evident sinonasal lesions, it is still possible to determine an increased incidence of morphofunctional changes in subjects occupationally exposed to wood dust. Our findings may lead to the identification of occupational groups prone to elevated risk of the disease. PMID- 11936914 TI - Epithelial cells in nasal fluids from patients with allergic rhinitis: how do they relate to epidermal growth factor, eosinophils and eosinophil cationic protein? AB - The nasal epithelium protects the underlying tissue from damage. Epithelial cell growth is controlled by epidermal growth factor (EGF) and is possibly affected by toxic proteins, e.g. eosinophil cationic protein (ECP). The aims of this study were to examine nasal fluid epithelial cell counts and their relations to EGF, eosinophils and ECP in 23 patients with seasonal allergic rhinitis and 20 healthy controls. Nasal fluid epithelial cell counts were lower in patients than in controls. EGF levels did not differ between patients and controls, and correlated with epithelial cell counts in controls but not in patients. Eosinophils and ECP were higher in patients than in controls, but did not correlate with epithelial cell counts. The role of growth factors, such as EGF, in regulating epithelial cells merits further study. PMID- 11936915 TI - Neutrophils are hyperactive in recurrent tonsillitis. AB - The pathogenesis of recurrent tonsillitis has not been fully explored. Most studies in this field have focused on pathogenic bacteria whereas less research has been done concerning the host defense. In earlier studies it was shown that there is an active cellular defense in the tonsillar surface secretion, consisting of phagocytes, and therefore in this study the possibility that this defense is altered in patients with recurrent tonsillitis was explored. Neutrophils were obtained from tonsillar surface secretions in eight patients with recurrent tonsillitis and eight healthy volunteers and the capacity of the neutrophils to respond to chemotactic stimuli was examined in an in vitro system. A significantly higher fraction of the neutrophils obtained from the patients with recurrent tonsillitis responded to chemotactic stimuli compared to those obtained from the healthy volunteers. It is concluded that there is a hyperactive cellular defense in the tonsillar surface secretion in patients with recurrent tonsillitis. This finding and its significance in the pathogenesis of recurrent tonsillitis are discussed. PMID- 11936916 TI - Bacteria and pathogenesis of human salivary calculus. AB - The exact cause of salivary calculus formation is unknown and the aim of this study was to ascertain whether bacteria play a role. Sialoliths from nine patients with chronic obstructive sialadenitis of the submandibular gland were analysed. Bacterial gene fragments were amplified from DNA extracted from salivary calculi by means of polymerase chain reaction using a universal bacterial primer pair. Comparative 16S ribosomal RNA sequence analysis was used for identification. We detected and identified oral bacteria (predominantly Streptococcus species) in all samples. The present results suggest a potential role for bacteria in the etiopathogenesis of sialolithiasis. PMID- 11936917 TI - Esthesioneuroblastoma and cervical lymph node metastases: clinical and therapeutic implications. PMID- 11936918 TI - Parastomal recurrence: a therapeutic challenge. PMID- 11936919 TI - Management of N3 neck. PMID- 11936920 TI - Differential expression and topography of adhesion molecules in laryngeal and oropharyngeal carcinomas. AB - This work describes the different patterns of expression of integrins and extracellular matrix proteins in normal and transformed mucosa in laryngeal and oropharyngeal carcinomas. Samples from each tumor group were sectioned and examined by immunohistochemistry using monoclonal antibodies raised against integrin chains (alpha2, alpha3, alpha6, beta1 and beta4) and their ligands (laminins 1 and 5, collagen type IV and two fibronectin isoforms: ED-A and ED-B). Controls were provided by samples of tumor-free laryngeal and oropharyngeal mucosa that had been removed during the surgical procedure. We found that the known distinct topographical pattern of integrins and the continuity of basement membrane components was altered in both groups but that the extent of changes was significantly more marked in oropharyngeal tumors, which are known to be more infiltrating and diffusive and to have a bad prognosis. These molecular patterns of expression can be used as an additional prognostic factor as they suggest a greater biological tumor aggressiveness of oropharyngeal tumors. We suggest that performing immunohistochemical analysis on biopsy samples may help in selecting the correct therapeutic strategy for these tumors and enable more accurate follow up. The above-mentioned molecules may become part of the diagnostic toolbox of head and neck surgical pathologists. PMID- 11936922 TI - Gavriel A. Ilizarov--Russian surgeon develops the external fixator. PMID- 11936923 TI - Screening for renal artery stenosis: is it justified? PMID- 11936924 TI - Incidental renal artery stenosis among a prospective cohort of hypertensive patients undergoing coronary angiography. AB - OBJECTIVE: To determine the feasibility, safety, and clinical yield of angiographic screening among hypertensive patients undergoing coronary angiography. PATIENTS AND METHODS: This study was a prospective cohort analysis of hypertensive patients who underwent cardiac catheterization at a tertiary care referral center from July 1998 to March 1999. Abdominal aortography was performed to screen for renal artery stenosis, the percentage of which was measured. RESULTS: The mean +/- SD age of the 297 study patients was 64.9+/-10.2 years; 58.6% were male, and 98.0% were white. Mean +/- SD systolic/diastolic blood pressure was 142.8+/-22.5/79.6+/-11.4 mm Hg. Aortography required a mean incremental dose of 62+/-9 mL of nonionic contrast agent. No complications were attributable to aortography. Of 680 renal arteries, 611 (90%) were visualized adequately. Also, 53% of patients had normal renal arteries, 28% had stenoses less than 50%, and 19.2% had stenoses of 50% or more. Renal artery stenosis was bilateral in 3.7% of patients and high grade (>70% stenosis) in 7%. Patients with renal artery stenosis were more likely to have had a previous coronary intervention. In multivariate analysis, systolic blood pressure (odds ratio [OR], 1.2; 95% confidence interval [CI], 1.03-138; P=.02), history of stroke or transient ischemic attack (OR, 2.7; 95% CI, 1.27-5.78; P=.01), and cancer (OR, 2.0; 95% CI, 1.02-3.82; P=.04) independently correlated with renal artery stenosis of 50% or more. CONCLUSION: The prevalence of incidental renal artery stenosis among hypertensive patients undergoing coronary catheterization is significant. Therefore, screening abdominal aortography should be considered in these patients to better define their risk of cardiovascular complications. PMID- 11936925 TI - Injuries in youth football: a prospective observational cohort analysis among players aged 9 to 13 years. AB - OBJECTIVE: To determine the risk of injury in youth football games. SUBJECTS AND METHODS: Nine hundred fifteen players aged 9 to 13 years on 42 teams participated, including 10 teams in each grade from grades 4 through 6 and 6 teams each in grades 7 and 8. The study was conducted in the fall of 1997. Injury incidence, prevalence, and severity were calculated for each grade level and player position. Additional analyses examined the number of injuries according to body weight. RESULTS: A total of 55 injuries occurred in games during the entire season (overall prevalence, 5.97%). Most injuries were mild, and the most common type was contusion, which occurred in 33 players (60%). Four injuries (7%) were severe enough to prevent players from participating for the rest of the season. All 4 severe injuries were fractures involving the ankle physis. The risk of injury increased as players matured in age and grade level. Injury risk for an eighth-grade player was 4 times greater than the risk of injury to a fourth-grade player. A trend was identified for heavier players to be at increased risk, but no significant correlation was evident between body weight and injury. CONCLUSION: Our prospective observational analysis showed that most youth football injuries are mild. Older and heavier players appear to be at higher risk. PMID- 11936926 TI - Chronic thrombotic microangiopathy associated with antineoplastic therapy with minimal hematologic effects. AB - OBJECTIVE: To describe 6 patients who developed progressive renal failure and renal thrombotic microangiopathy (TM) not accompanied by the characteristic hematologic disturbances of TM syndromes. PATIENTS AND METHODS: Portions of renal biopsy specimens from each patient were examined by light and electron microscopy for histopathologic evidence of TM. Antecedent clinical events, laboratory evidence of hemolysis and thrombocytopenia, and clinical outcome were documented. Medical records were reviewed and clinical data, including laboratory values, treatment, and outcome, were recorded. RESULTS: In each case, a slowly progressive uremia evolved after radiation and/or chemotherapy without laboratory evidence of acute hemolysis or thrombocytopenia. Renal biopsy specimens in all cases showed TM and tubulointerstitial scarring, suggesting both acute and chronic renal injury. Two of the 6 patients underwent plasma exchange therapy without improvement of renal function. Three patients treated with angiotensin converting enzyme inhibitors for coexisting systemic hypertension remained stable or had mild improvement in renal function. CONCLUSIONS: A small subset of patients treated for malignancy developed slowly evolving uremia associated with renal TM without marked hematologic abnormalities. In the absence of thrombocytopenia and other typical laboratory findings, the diagnosis of renal TM may be overlooked. PMID- 11936927 TI - Computed tomography of benign intrapulmonary lymph nodes: retrospective comparison with sarcoma metastases. AB - OBJECTIVE: To determine whether the computed tomographic (CT) characteristics of benign intrapulmonary lymph nodes and small sarcoma metastases are sufficiently characteristic to allow specific prospective identification. PATIENTS AND METHODS: Preoperative chest CT scans of 41 patients with benign intrapulmonary lymph nodes and 33 patients with sarcoma metastases seen from 1991 through 1996 were retrospectively reviewed and correlated with pathologic findings. RESULTS: Fifty-seven benign intrapulmonary lymph nodes were found. Twenty-six (46%) were subpleural, 38 (67%) were oval, and 46 (81%) were located in the lower portions of the lungs; 43 (75%) had a lymphatic distribution on CT and 54 (95%) at pathologic review. Ninety-eight sarcoma metastases were found. Thirteen (13%) were subpleural, 15 (15%) were oval, and 56 (57%) were in the lower portions of the lungs; 29 (30%) had a lymphatic distribution on CT and 45 (46%) at pathologic review. CONCLUSION: Benign intrapulmonary lymph nodes were more likely than sarcoma metastases to be oval, to occur in a lymphatic distribution, and to be located subpleurally. PMID- 11936928 TI - Barriers to osteoporosis identification and treatment among primary care physicians and orthopedic surgeons. AB - OBJECTIVE: To understand better the barriers among orthopedic surgeons and primary care physicians in identifying and treating possible osteoporosis in patients hospitalized with a fragility fracture sustained spontaneously or from a fall no greater than standing height. METHODS: A 1-page, 7-question survey was sent to 35 admitting orthopedic surgeons and 75 primary care physicians at a midwestern managed care organization in March 2001. Returned surveys were collected until 30 days had passed since the mailing. Primary care physicians were board-certified family practitioners and internal medicine physicians. All orthopedists were admitting surgeons in the hospital system. Responders were anonymous, and posted surveys were returned to the Orthopaedic Collaborative Practice office. The surveys were color-coded to separate responses from orthopedic surgeons and primary care physicians. RESULTS: Thirty-one surveys were returned: 23 (31%) from primary care physicians and 8 (23%) from orthopedic surgeons. Survey respondents agreed that the responsibility for postfracture attention to nutritional needs, including calcium and vitamin D, rested with the primary care provider. When asked about barriers to recommending bone mineral density testing with dual energy x-ray absorptiometry, 9 primary care physicians (39%) thought this type of testing was unnecessary for treatment, and 4 primary care physicians (17%) thought a barrier was caused by patient frailty. Primary care physicians indicated that potential adverse effects of medication (n=14 [61%]) and cost of therapy (n=13 [57%]) were the main factors limiting treatment. When asked to identify the single most important barrier in treatment, 14 physicians (61%) indicated cost was the greatest deterrent. Twenty-one primary care physicians (91%) reported they would be more likely to treat a patient with osteoporosis if a safe medication with proven fracture risk reduction were available. Primary care physicians indicated they were more likely to treat independently living adults (n=12 [52%]) and women compared with men (n=15 [65%]). All orthopedic surgeons (n=8) were willing for all patients to be evaluated in consultation with a nurse practitioner. Primary care respondents were less apt to agree with a nurse practitioner referral (n=5 [22%]). Both primary care physicians (n=16 [70%]) and orthopedic surgeons (n=4 [50%]) agreed that there is a need for increased primary care education about managing osteoporosis in patients hospitalized with low-impact fracture. CONCLUSIONS: Orthopedic surgeons were consistent in their opinion that postfracture attention to osteoporosis should rest with the primary care physician. Primary care physicians agree but report that cost and possible adverse effects of medication are major barriers to this care. Despite therapies for high-risk postfracture patients showing relative safety and proven efficacy in reducing future fractures, deterrents to this care are focused on cost and potential adverse effects. Further education is needed to promote a standard of care for the postfracture patient that is directed toward the prevention of a subsequent fracture. PMID- 11936929 TI - Challenges in the development of effective peptide vaccines for cancer. AB - The ability of the immune system to recognize malignant cells has opened the door to development of tumor vaccines to treat or prevent various types of cancer. In the era of molecular biology, the tumor antigens recognized by the immune system have been identified, allowing the generation of subunit vaccines that may improve safety and efficacy compared with more crude vaccines such as irradiated tumor cells and tumor cell lysates. Synthetic peptides corresponding to defined antigenic epitopes for tumor-reactive lymphocytes represent one of the new types of vaccines currently being developed to treat or prevent various types of malignant disorders. The design of peptide-based vaccines to stimulate antitumor T-cell responses has many attractive features such as ease of manufacturing and characterization (ie, quality control), as well as an excellent safety profile in past clinical studies. However, ambiguous results from initial clinical trials indicate that these vaccines are far from optimal and that considerable efforts for their optimization lie ahead. We attempt to address the 8 most important challenges we currently face for developing peptide-based vaccines that would effectively induce immune responses leading to antitumor effects. PMID- 11936930 TI - Use of intranasal cromolyn sodium for allergic rhinitis. AB - Allergic rhinitis affects 10% to 20% of Americans. It frequently coexists with other conditions, such as allergic conjunctivitis, sinusitis, and asthma, and is associated with impaired occupational function and performance in school, decreased quality of life, and increased health care costs. An efficacious agent with minimal adverse effects and a lack of drug interactions is needed to help simplify treatment of allergic rhinitis, especially in patients with comorbidities. Controlled studies of intranasal cromolyn sodium therapy for patients with seasonal and perennial allergic rhinitis are reviewed, and appropriate candidates for treatment with this agent are discussed. Cromolyn inhibits the degranulation of sensitized mast cells, thereby blocking the release of inflammatory and allergic mediators. It reduces symptoms of allergic rhinitis, and, when used prophylactically, cromolyn can prevent symptoms from occurring. Controlled studies comparing cromolyn with placebo, intranasal corticosteroids, and antihistamines have shown the efficacy of cromolyn in relieving rhinitis symptoms. In addition, because cromolyn is poorly absorbed systemically, it is well tolerated and not associated with drug interactions. Intranasal cromolyn has an excellent safety record, is available as an over-the-counter medication, and has been proved to be efficacious in patients with allergic rhinitis. PMID- 11936931 TI - Identifying and counseling women at increased risk for breast cancer. AB - Women at increased risk for breast cancer should be identified and counseled about options for risk reduction. Identifying such women is simplified with use of the National Cancer Institute Risk Assessment tool, a computer-based tool that incorporates information on 6 risk factors for estimating an individual's risk of developing breast cancer. However, the tool does not incorporate all known or possible risk factors and may underestimate risk, particularly among women with a complex family history of breast cancer for whom alternative models of risk assessment are more appropriate. Women found to have an increased risk of breast cancer should be counseled about options for management, including close surveillance, lifestyle modifications, chemoprevention with tamoxifen, enrollment in a breast cancer prevention clinical trial, and prophylactic mastectomy and/or oophorectomy. In the absence of consensus about which risk level is best suited to which option, decisions about risk reduction depend as much on an individual's priorities and risk aversion as on numerical risk estimates. PMID- 11936933 TI - 61-year-old man with dyspnea and bilateral foot drop. PMID- 11936934 TI - Assessing clinical significance in measuring oncology patient quality of life: introduction to the symposium, content overview, and definition of terms. AB - The Clinical Significance Consensus Meeting Group of the Symposium on the Clinical Significance of Quality-of-Life Measures in Cancer Patients produced 6 articles regarding the clinical significance of quality of life (QOL) assessments in oncology. The 6 articles deal with the methods used to date: group-vs individual clinical significance; single items vs summated scores; patient, clinician, and population perspectives; assessment of changes over time; and communication of results. The articles were produced by a team of 30 QOL research experts assembled in a consensus writing meeting held at the Mayo Clinic, Rochester, Minn, October 6 and 7, 2000. This introduction describes the need for the articles, definitions of key terms, and plans for the future. It is hoped that this series of articles will serve as a resource for individuals conducting cancer QOL research and for clinicians considering incorporation of QOL assessment in the treatment of patients with neoplastic diseases. A secondary goal is to stimulate further discussion and research in the interpretation of QOL assessments. PMID- 11936935 TI - Methods to explain the clinical significance of health status measures. AB - One can classify ways to establish the interpretability of quality-of-life measures as anchor based or distribution based. Anchor-based measures require an independent standard or anchor that is itself interpretable and at least moderately correlated with the instrument being explored. One can further classify anchor-based approaches into population-focused and individual-focused measures. Population-focused approaches are analogous to construct validation and rely on multiple anchors that frame an individual's response in terms of the entire population (eg, a group of patients with a score of 40 has a mortality of 20%). Anchors for population-based approaches include status on a single item, diagnosis, symptoms, disease severity, and response to treatment. Individual focused approaches are analogous to criterion validation. These methods, which rely on a single anchor and establish a minimum important difference in change in score, require 2 steps. The first step establishes the smallest change in score that patients consider, on average, to be important (the minimum important difference). The second step estimates the proportion of patients who have achieved that minimum important difference. Anchors for the individual-focused approach include global ratings of change within patients and global ratings of differences between patients. Distribution-based methods rely on expressing an effect in terms of the underlying distribution of results. Investigators may express effects in terms of between-person standard deviation units, within person standard deviation units, and the standard error of measurement. No single approach to interpretability is perfect. Use of multiple strategies is likely to enhance the interpretability of any particular instrument. PMID- 11936936 TI - Group vs individual approaches to understanding the clinical significance of differences or changes in quality of life. AB - This article focuses on the traversing of group and individual levels of quality of-life data. A deductive approach is used to address the extent to which group data can be used to estimate clinical significance at the individual level. An inductive approach is used to evaluate the extent to which individual change data can be brought to the group level to define clinical significance. Both approaches have benefits and drawbacks. This article addresses how clinical significance can be defined for an individual when the threshold for meaningfulness is drawn from group data. It also addresses the condition under which one can use the same threshold difference for group vs individual differences or changes. A sample inductive approach explores the means to identify a clinically significant result or change, with use of insights from cognitive psychology. In most deductive approaches, the identification of a clinically significant difference or change requires identification of a criterion (or at least an interpretable anchor) against which the significance of a change in respondent score is compared. PMID- 11936937 TI - Mycobacterium bovis vertebral osteomyelitis as a complication of intravesical BCG use. AB - We report a culture-proven case of Mycobacterium bovis vertebral osteomyelitis in a 76-year-old man who had undergone intravesical BCG therapy for bladder cancer 7 years previously. He presented with debilitating back pain and had radiographic evidence of T6-7 disk space destruction with involvement of adjacent vertebrae. Tissue culture from the disk space confirmed the diagnosis of vertebral osteomyelitis due to hematogenous spread of M. bovis. Treatment with antituberculous medications was begun soon after tissue diagnosis was made, and the patient fared well with medical therapy alone. Although uncommon, this infectious complication of BCG therapy should always be considered in the appropriate clinical setting. Timely diagnosis is important, because chemotherapy, when initiated early in the disease, can preclude the necessity for surgical intervention. PMID- 11936938 TI - Hypothyroidism-related CA 19-9 elevation. PMID- 11936939 TI - Autonomic tone and benefits of cardiac rehabilitation programs. PMID- 11936940 TI - Telehealth: promise or peril? AB - As you walk down the hall of the clinic, you see through the window millions of jewels as the sun is reflected off the ice on the Bering Sea. You enter the consultation room and greet your patient, Missy Becker. You have arranged for a consult with Dr. Sarah McKinnon about a lesion that Missy has. Turning on the computer you tell Missy that Dr. McKinnon, a dermatologist in Chicago, will be examining her today. When the computer monitor comes alive, you say hello to Dr. McKinnon and introduce her to Missy. You set up the camera that has finger like probes so that Dr. McKinnon can manipulate both the camera and probes. Using these touch-sensitive probes she examines Missy s lesion just as she would if she were physically with you in the consultation room. "Good news," Dr. McKinnon says after a thorough examination, "As far as I can tell now, the lesion is benign. Do a biopsy and put it under the telemicroscope so I can see it." The fact that Dr. McKinnon is in Chicago and you and Missy are separated by over 3,000 miles does not even evoke a comment. PMID- 11936941 TI - Telemedicine: follow the money modalities. AB - Until the late 1990s,e private and public third-party payers generally did not have explicit policies to pay for telehealth services. The Balanced Budget Act of 1997 signaled a significant change in Medicare payment policies opening the door for telemedicine reimbursement. This article traces the development of current Medicare telemedicine payment policies, beginning with the BBA of 1997 and including current medicare payment legislation. Issues related to telemedicine payment by both Medicare and other third party payers are presented; implication for the future, and the role of the nursing community are discussed. PMID- 11936942 TI - Health information privacy protection: crisis or common sense? AB - Concerns about the protection of personally identifiable information are not unique to the health care industry; however, consumers view their medical records as more "private" than other information, such as financial data, because involuntary disclosure can affect jobs or health insurance status. This paper briefly touches upon new sweeping federal privacy standards mandated under the Health Insurance Portability and Accountability Act of 1996 (HIPAA). The article outlines who and what is covered under the new rules, considers how practitioners can approach compliance with common sense, addresses concerns related to risk management, discusses consumer health privacy issues, and notes the difficulty of evaluating these rules and regulations. The article also looks at some unique privacy issues facing telemedicine and telehealth practitioners. PMID- 11936943 TI - Legal considerations for nurses practicing in a telehealth setting. AB - During the last century the world has become increasingly reliant on a variety of technologies to manage information needs. Escalation in deployment of remote technology to enhance health care, accompanied by expanded public and private reimbursement for distant care, indicates increasing acceptance of these technologies. Yet many legal and regulatory questions regarding the provision of health care using these technologies remain. This article will discuss the foundations of telehealth nursing and address legal and regulatory issues emerging from this new practice modality. PMID- 11936944 TI - Window of opportunity for home care nurses: telehealth technologies. AB - Multiple telehealth technologies are currently available for nurses working in home care and other community settings. Furthermore, the rate at which new electronic technologies, which can promote home care nursing, are introduced continues to increase. This article addresses the need for increased use of information technologies in home care nursing, describes how information technologies are changing the way we give nursing care, provides examples of telehealth nursing in home care, and discusses the future of telehealth technologies in nursing. PMID- 11936945 TI - The nursing shortage and ethics: up front and personal. AB - In times of crisis it is often that values and beliefs and, subsequently, our ethics are challenged. Authors in OJIN have addressed some of the ethical issues that nurses face in relation to crises in health care. For example, P. J. Maddox (1998) addressed ethical issues surrounding economic constraints and scarce resources. In this current issue of OJIN, Cheryl Peterson (2001) writes of the ethical dilemma nurses face in trying to comply with the Code for Nurses with Interpretive Statement, 1985 in light of the present nursing shortage. We invite you to read these and the other OJIN articles on the shortage and then submit to this column a story about how the nursing shortage has ethically affected you or your patients/clients. PMID- 11936946 TI - Using health statistics: a Nightingale legacy. AB - No more forceful example of the value of using health statistics to understand and improve health conditions exists than displayed by Florence Nightingale. The recent book by Dossey (1999), Florence Nightingale: Mystic, Visionary, Healer, relates the dramatic tale of Nightingale s use of statistics to understand the causes of deaths in the Crimean War and of her advocacy to standardize the collection of medical data within the army and in civilian hospitals. For her, the use of health statistics was a major tool to improve health and influence public opinion. PMID- 11936947 TI - Nursing interventions revisited. AB - Medication administration is one of the most highly contested tasks delegated by registered nurses to others. Readers are encouraged to complete this brief survey asking about which nursing interventions must be performed by an RN versus those that can be safely delegated. PMID- 11936948 TI - Establishing a nursing student learning center for women's reproductive health in Nepal. AB - The goal of this paper is to describe the establishment of a self-sustaining Student Learning Center (SLC) employing humanistic anatomical models to aid in the teaching of family planning and reproductive health clinical skills to nursing students in Nepal. In response to a local needs assessment a plan was developed to implement more thorough training of nursing students in family planning clinical skills. Nepalese nursing leaders worked together with a U.S. Non-Governmental Organization (NGO) to implement this project. This paper details the need for the SLCs, the content and structure of the SLCs, and the process of establishing an SLC at Tribhuvan University, Maharjgunj Nursing Campus in Kathmandu, the largest of all nursing campuses in Nepal. PMID- 11936949 TI - State boards of nursing and scope of practice of registered nurses performing complementary therapies. AB - This article provides a report of State Boards of Nursing (BONs) policies on the use of complementary therapies by registered nurses. This investigation was conducted for the White House Commission on Complementary Alternative Medicine Policy. The target sample for this report was fifty-three BONs in the United States. Forty-seven percent of the BONs had taken positions that permitted nurses to practice a range of complementary therapies; thirteen percent were in the process of discussing this matter; and forty percent, although they had not formally addressed the topic, did not necessarily discourage these practices. The results of this study can be used to encourage more discussion among the BONs and within states about nurses practice of complementary therapies. Nurses are encouraged to become aware of their state s position regarding complementary therapy and to facilitate the integration of complementary therapies into their work environment. PMID- 11936950 TI - Oncostatin M induced alpha1-antitrypsin (AAT) gene expression in Hep G2 cells is mediated by a 3' enhancer. AB - alpha(1)-Antitrypsin (AAT) is the major serine proteinase inhibitor (SERPIN A1) in human plasma. Its target proteinase is neutrophil elastase and its main physiological function is protection of the lower respiratory tract from the destructive effects of neutrophil elastase during an inflammatory response. Circulating levels of AAT rise 2-3-fold during inflammation and the liver produces most of this increase. The cytokines oncostatin M (OSM) and interleukin 6 have been shown to be mainly responsible for this effect, which is mediated via the interaction of cytokine-inducible transcription factors with regulatory elements within the gene. In the present study, we report for the first time that OSM stimulation of hepatocyte AAT occurs via an interaction between the hepatocyte promoter and an OSM-responsive element at the 3'-end of the AAT gene. This effect is mediated by the transcription factor signal transducer and activator of transcription 3 ('STAT 3') binding to an OSM-responsive element (sequence TTCTCTTAA), and this site is distinct from, but close to, a previously reported interleukin-6-responsive element. PMID- 11936951 TI - Function of the 90-loop (Thr90-Glu100) region of staphylokinase in plasminogen activation probed through site-directed mutagenesis and loop deletion. AB - Staphylokinsae (SAK) forms a bimolecular complex with human plasmin(ogen) and changes its substrate specificity by exposing new exosites that enhances accession of substrate plasminogen (PG) to the plasmin (Pm) active site. Protein modelling studies indicated the crucial role of a loop in SAK (SAK 90-loop; Thr(90)-Glu(100)) for the docking of the substrate PG to the SAK-Pm complex. Function of SAK 90-loop was studied by site-directed mutagenesis and loop deletion. Deletion of nine amino acid residues (Tyr(92)-Glu(100)) from the SAK 90 loop, resulted in approximately 60% reduction in the PG activation, but it retained the ability to generate an active site within the complex of loop mutant of SAK (SAKDelta90) and Pm. The preformed activator complex of SAKDelta90 with Pm, however, displayed a 50-60% reduction in substrate PG activation that remained unaffected in the presence of kringle domains (K1+K2+K3+K4) of PG, whereas PG activation by SAK-Pm complex displayed approximately 50% reduction in the presence of kringles, suggesting the involvement of the kringle domains in modulating the PG activation by native SAK but not by SAKDelta90. Lysine residues (Lys(94), Lys(96), Lys(97) and Lys(98)) of the SAK 90-loop were individually mutated into alanine and, among these four SAK loop mutants, SAK(K97A) and SAK(K98A) exhibited specific activities about one-third and one-quarter respectively of the native SAK. The kinetic parameters of PG activation of their 1:1 complex with Pm indicated that the K(m) values of PG towards the activator complex of these two SAK mutants were 4-6-fold higher, suggesting the decreased accessibility of the substrate PG to the activator complex formed by these SAK mutants. These results demonstrated the involvement of the Lys(97) and Lys(98) residues of the SAK 90-loop in assisting the interaction with substrate PG. These interactions of SAK-Pm activator complex via the SAK 90-loop may provide additional anchorage site(s) to the substrate PG that, in turn, may promote the overall process of SAK-mediated PG activation. PMID- 11936952 TI - Total direct chemical synthesis and biological activities of human group IIA secretory phospholipase A2. AB - Human group IIA secretory phospholipase A(2) (hGIIA sPLA(2)) is reported to be involved in inflammation, since its expression level is enhanced under various inflammatory conditions. In this work, we report the total chemical synthesis of this enzyme (124 amino acids) by solid-phase method. The identity of the protein, in denatured or folded (7 disulphide bonds) forms, was confirmed by electrospray MS. Synthetic sPLA(2) possesses the same circular dichroism spectrum, enzymic activity in hydrolysing different phospholipid substrates, and inhibitory effect in thrombin formation from prothrombinase complex as the recombinant sPLA(2). Furthermore, LY311727, a reported specific hGIIA sPLA(2) inhibitor, is able to inhibit the synthetic and the recombinant enzymes with the same efficiency. This study demonstrates that chemically continuous solid phase synthesis is an alternative and less time-consuming approach to producing small, structurally folded and fully active proteins of up to 124 amino acids, such as hGIIA sPLA(2). Moreover, this technique provides more flexibility in analogue synthesis to elucidate their physiological functions and pathological effects. PMID- 11936953 TI - Metabolism of the novel Ca2+-mobilizing messenger nicotinic acid-adenine dinucleotide phosphate via a 2'-specific Ca2+-dependent phosphatase. AB - Nicotinic acid-adenine dinucleotide phosphate (NAADP) is a newly described Ca2+ mobilizing nucleotide that appears to target intracellular Ca2+-release channels distinct from those sensitive to inositol trisphosphate or ryanodine/cyclic ADP ribose. Little, however, is known concerning the regulation of cellular NAADP levels. In the present study, we have characterized the metabolism of NAADP by brain membranes. From HPLC and MS analyses we show that loss of NAADP was associated with the appearance of a major product that is likely to be nicotinic acid-adenine dinucleotide (NAAD), the dephosphorylated form of NAADP. Dephosphorylation of NAADP, but not 3'-NAADP, was dramatically attenuated by Ca2+ chelators and stimulated by Ca2+ over a physiological range in a calmodulin insensitive manner. In contrast, NADP was metabolized predominantly to ADP-ribose phosphate via glycohydrolase activity, although slower Ca2+-dependent dephosphorylation of both NADP and 2'-AMP could also be demonstrated. This is the first report describing a Ca2+-regulated 2'-specific phosphatase which is probably the major pathway for the inactivation of NAADP in brain. Our data provide a potential feedback mechanism for limiting NAADP-induced Ca2+ release within cells through stimulation of NAADP metabolism by Ca2+ and strongly support a signalling role for this novel nucleotide in the brain. PMID- 11936954 TI - Mitochondria-dependent and -independent mechanisms in tumour necrosis factor related apoptosis-inducing ligand (TRAIL)-induced apoptosis are both regulated by interferon-gamma in human breast tumour cells. AB - Tumour necrosis factor-related apoptosis-inducing ligand (TRAIL/APO-2L) induces apoptosis in a variety of tumour cells upon binding to death receptors TRAIL-R1 and TRAIL-R2. Here we describe the sensitization by interferon (IFN)-gamma to TRAIL-induced apoptosis in the breast tumour cell lines MCF-7 and MDA-MB231. IFN gamma promoted TRAIL-mediated activation of caspase-8, Bcl-2 interacting domain death agonist (Bid) degradation, Bcl-2-associated X protein (Bax) translocation to mitochondria, cytochrome c release to the cytosol and activation of caspase-9 in these cell lines. No changes in the expression of TRAIL receptors were observed upon IFN-gamma treatment. Overexpression of Bcl-2 in MCF-7 cells completely inhibited IFN-gamma-induced sensitization to TRAIL-mediated cell death. Interestingly, TRAIL-induced apoptosis was also clearly enhanced by IFN gamma in caspase-3-overexpressing MCF-7 cells, in the absence of Bax translocation to mitochondria and cytochrome c release to the cytosol. In summary, our results suggest that IFN-gamma facilitates TRAIL-induced activation of mitochondria-regulated as well as mitochondria-independent apoptotic pathways in breast tumour cells. PMID- 11936955 TI - Sources of variability and effect of experimental approach on expression profiling data interpretation. AB - BACKGROUND: We provide a systematic study of the sources of variability in expression profiling data using 56 RNAs isolated from human muscle biopsies (34 Affymetrix MuscleChip arrays), and 36 murine cell culture and tissue RNAs (42 Affymetrix U74Av2 arrays). RESULTS: We studied muscle biopsies from 28 human subjects as well as murine myogenic cell cultures, muscle, and spleens. Human MuscleChip arrays (4,601 probe sets) and murine U74Av2 Affymetrix microarrays were used for expression profiling. RNAs were profiled both singly, and as mixed groups. Variables studied included tissue heterogeneity, cRNA probe production, patient diagnosis, and GeneChip hybridizations. We found that the greatest source of variability was often different regions of the same patient muscle biopsy, reflecting variation in cell type content even in a relatively homogeneous tissue such as muscle. Inter-patient variation was also very high (SNP noise). Experimental variation (RNA, cDNA, cRNA, or GeneChip) was minor. Pre-profile mixing of patient cRNA samples effectively normalized both intra- and inter patient sources of variation, while retaining a high degree of specificity of the individual profiles (86% of statistically significant differences detected by absolute analysis; and 85% by a 4-pairwise comparison survival method). CONCLUSIONS: Using unsupervised cluster analysis and correlation coefficients of 92 RNA samples on 76 oligonucleotide microarrays, we found that experimental error was not a significant source of unwanted variability in expression profiling experiments. Major sources of variability were from use of small tissue biopsies, particularly in humans where there is substantial inter-patient variability (SNP noise). PMID- 11936956 TI - First isolation of the enterohaemorrhagic Escherichia coli O145:H- from cattle in feedlot in Argentina. AB - BACKGROUND: Enterohaemorrhagic Escherichia coli (EHEC) is considered to be common cause of haemorrhagic colitis (HC), thrombotic thrombocytopenic purpura and haemolytic uraemic syndrome (HUS) in humans. In a previous paper, we have demonstrated that EHEC are commonly found in the intestines of livestock. Infections in humans are, in part, a consequence of consumption of undercooked meat or raw milk. Argentina has one of the highest records of HUS (300-400 cases/year; 22/100,000 children under 4 years of age). The aim of this work is to communicate the first isolation of O145:H-from cattle in this country and characterize the virulence cassette, providing useful information to evaluate the risk of foodborne transmission of this emergent non-O157:H7 serotype. RESULTS: EHEC O145:H- was isolated from cattle in an Argentinian feedlot. Pheno- and genotype of nine strains were characterized, corresponding to several virulence cassettes: VT2+eaeA+ Mp+ (n = 5), VT2+eaeA+ (n = 1), VT1+eaeA+ Mp+ (n = 2), and VT1+eaeA+ (n = 1). Strains isolated from the same animal were considered only when they showed a different virulence pattern. The clonal relationship was studied by RAPD. Strains were distributed in two RAPD profiles, which corresponded to the presence of either, VT1+ or VT2+ genotype. No difference was detected by RAPD analysis between Mp+ or Mp- strains. CONCLUSIONS: This was the first isolation of EHEC O145:H- serotype in Argentina enlarging the list of non O157:H7 serotypes isolated from cattle in this country by us. All O145:H-strains carried several virulence factors which allow us to predict their potential ability to develop haemolytic uraemic syndrome in humans. PMID- 11936957 TI - Effect of interlukin-1beta on proliferation of gastric epithelial cells in culture. AB - BACKGROUND: Helicobacter pylori is the main risk factor for the development of non-cardia gastric cancer. Increased proliferation of the gastric mucosa is a feature of H. pylori infection. Mucosal interkeukin-1beta production is increased in H. pylori infection and IL-1beta genotypes associated with increased pro inflammatory activity are risk factors for the development of gastric cancer. The effect of IL-1beta on gastric epithelial cell proliferation has been examined in this study. METHODS: AGS cells were cultured with IL-1beta. DNA synthesis was assed by [3H]thymidine incorporation and total viable cell numbers by MTT assay. RESULTS: IL-1beta dose dependently increased DNA synthesis and cell numbers. The enhanced proliferation was blocked by interleukin-1 receptor antagonist. Addition of neutralising antibody to GM-CSF reduced IL-1beta-stimulated proliferation by 31 +/- 4 %. GM-CSF alone significantly stimulated proliferation. Addition or neutralisation of IL-8 had no effect on basal or IL-1beta-stimulated proliferation. The tyrosine kinase inhibitor genistein completely blocked IL 1beta-stimulated proliferation and inhibition of the extracellular signal related kinase pathway with PD 98059 inhibited IL-1beta stimulated proliferation by 58 +/ 5 %. CONCLUSIONS: IL-1beta stimulates proliferation in gastric epithelial cells. Autocrine stimulation by GM-CSF contributes to this proliferative response. Signalling via tyrosine kinase activity is essential to the mitogenic response to IL-1beta. The extracellular signal related kinase pathway is involved in, but not essential to downstream signalling. IL-1beta may contribute to the hyperproliferation seen in H. pylori- infected gastric mucosa, and be involved in the carcinogenic process. PMID- 11936959 TI - Slow feature analysis: unsupervised learning of invariances. AB - Invariant features of temporally varying signals are useful for analysis and classification. Slow feature analysis (SFA) is a new method for learning invariant or slowly varying features from a vectorial input signal. It is based on a nonlinear expansion of the input signal and application of principal component analysis to this expanded signal and its time derivative. It is guaranteed to find the optimal solution within a family of functions directly and can learn to extract a large number of decorrelated features, which are ordered by their degree of invariance. SFA can be applied hierarchically to process high dimensional input signals and extract complex features. SFA is applied first to complex cell tuning properties based on simple cell output, including disparity and motion. Then more complicated input-output functions are learned by repeated application of SFA. Finally, a hierarchical network of SFA modules is presented as a simple model of the visual system. The same unstructured network can learn translation, size, rotation, contrast, or, to a lesser degree, illumination invariance for one-dimensional objects, depending on only the training stimulus. Surprisingly, only a few training objects suffice to achieve good generalization to new objects. The generated representation is suitable for object recognition. Performance degrades if the network is trained to learn multiple invariances simultaneously. PMID- 11936958 TI - Selecting information technology for physicians' practices: a cross-sectional study. AB - BACKGROUND: Many physicians are transitioning from paper to electronic formats for billing, scheduling, medical charts, communications, etc. The primary objective of this research was to identify the relationship (if any) between the software selection process and the office staff's perceptions of the software's impact on practice activities. METHODS: A telephone survey was conducted with office representatives of 407 physician practices in Oregon who had purchased information technology. The respondents, usually office managers, answered scripted questions about their selection process and their perceptions of the software after implementation. RESULTS: Multiple logistic regression revealed that software type, selection steps, and certain factors influencing the purchase were related to whether the respondents felt the software improved the scheduling and financial analysis practice activities. Specifically, practices that selected electronic medical record or practice management software, that made software comparisons, or that considered prior user testimony as important were more likely to have perceived improvements in the scheduling process than were other practices. Practices that considered value important, that did not consider compatibility important, that selected managed care software, that spent less than 10,000 dollars, or that provided learning time (most dramatic increase in odds ratio, 8.2) during implementation were more likely to perceive that the software had improved the financial analysis process than were other practices. CONCLUSION: Perhaps one of the most important predictors of improvement was providing learning time during implementation, particularly when the software involves several practice activities. Despite this importance, less than half of the practices reported performing this step. PMID- 11936960 TI - Information loss in an optimal maximum likelihood decoding. AB - The mutual information between a set of stimuli and the elicited neural responses is compared to the corresponding decoded information. The decoding procedure is presented as an artificial distortion of the joint probabilities between stimuli and responses. The information loss is quantified. Whenever the probabilities are only slightly distorted, the information loss is shown to be quadratic in the distortion. PMID- 11936961 TI - The reliability of the stochastic active rotator. AB - The reliability of firing of excitable-oscillating systems is studied through the response of the active rotator, a neuronal model evolving on the unit circle, to white gaussian noise. A stochastic return map is introduced that captures the behavior of the model. This map has two fixed points: one stable and the other unstable. Iterates of all initial conditions except the unstable point tend to the stable fixed point for almost all input realizations. This means that to a given input realization, there corresponds a unique asymptotic response. In this way, repetitive stimulation with the same segment of noise realization evokes, after possibly a transient time, the same response in the active rotator. In other words, this model responds reliably to such inputs. It is argued that this results from the nonuniform motion of the active rotator around the unit circle and that similar results hold for other neuronal models whose dynamics can be approximated by phase dynamics similar to the active rotator. PMID- 11936962 TI - A proposed function for hippocampal theta rhythm: separate phases of encoding and retrieval enhance reversal of prior learning. AB - The theta rhythm appears in the rat hippocampal electroencephalogram during exploration and shows phase locking to stimulus acquisition. Lesions that block theta rhythm impair performance in tasks requiring reversal of prior learning, including reversal in a T-maze, where associations between one arm location and food reward need to be extinguished in favor of associations between the opposite arm location and food reward. Here, a hippocampal model shows how theta rhythm could be important for reversal in this task by providing separate functional phases during each 100-300 msec cycle, consistent with physiological data. In the model, effective encoding of new associations occurs in the phase when synaptic input from entorhinal cortex is strong and long-term potentiation (LTP) of excitatory connections arising from hippocampal region CA3 is strong, but synaptic currents arising from region CA3 input are weak (to prevent interference from prior learned associations). Retrieval of old associations occurs in the phase when entorhinal input is weak and synaptic input from region CA3 is strong, but when depotentiation occurs at synapses from CA3 (to allow extinction of prior learned associations that do not match current input). These phasic changes require that LTP at synapses arising from region CA3 should be strongest at the phase when synaptic transmission at these synapses is weakest. Consistent with these requirements, our recent data show that synaptic transmission in stratum radiatum is weakest at the positive peak of local theta, which is when previous data show that induction of LTP is strongest in this layer. PMID- 11936963 TI - Self-organization in the basal ganglia with modulation of reinforcement signals. AB - Self-organization is one of fundamental brain computations for forming efficient representations of information. Experimental support for this idea has been largely limited to the developmental and reorganizational formation of neural circuits in the sensory cortices. We now propose that self-organization may also play an important role in short-term synaptic changes in reward-driven voluntary behaviors. It has recently been shown that many neurons in the basal ganglia change their sensory responses flexibly in relation to rewards. Our computational model proposes that the rapid changes in striatal projection neurons depend on the subtle balance between the Hebb-type mechanisms of excitation and inhibition, which are modulated by reinforcement signals. Simulations based on the model are shown to produce various types of neural activity similar to those found in experiments. PMID- 11936964 TI - Population computation of vectorial transformations. AB - Many neurons of the central nervous system are broadly tuned to some sensory or motor variables. This property allows one to assign to each neuron a preferred attribute (PA). The width of tuning curves and the distribution of PAs in a population of neurons tuned to a given variable define the collective behavior of the population. In this article, we study the relationship of the nature of the tuning curves, the distribution of PAs, and computational properties of linear neuronal populations. We show that noise-resistant distributed linear algebraic processing and learning can be implemented by a population of cosine tuned neurons assuming a nonuniform but regular distribution of PAs. We extend these results analytically to the noncosine tuning and uniform distribution case and show with a numerical simulation that the results remain valid for a nonuniform regular distribution of PAs for broad noncosine tuning curves. These observations provide a theoretical basis for modeling general nonlinear sensorimotor transformations as sets of local linearized representations. PMID- 11936965 TI - Redistribution of synaptic efficacy supports stable pattern learning in neural networks. AB - Markram and Tsodyks, by showing that the elevated synaptic efficacy observed with single-pulse long-term potentiation (LTP) measurements disappears with higher frequency test pulses, have critically challenged the conventional assumption that LTP reflects a general gain increase. This observed change in frequency dependence during synaptic potentiation is called redistribution of synaptic efficacy (RSE). RSE is here seen as the local realization of a global design principle in a neural network for pattern coding. The underlying computational model posits an adaptive threshold rather than a multiplicative weight as the elementary unit of long-term memory. A distributed instar learning law allows thresholds to increase only monotonically, but adaptation has a bidirectional effect on the model postsynaptic potential. At each synapse, threshold increases implement pattern selectivity via a frequency-dependent signal component, while a complementary frequency-independent component nonspecifically strengthens the path. This synaptic balance produces changes in frequency dependence that are robustly similar to those observed by Markram and Tsodyks. The network design therefore suggests a functional purpose for RSE, which, by helping to bound total memory change, supports a distributed coding scheme that is stable with fast as well as slow learning. Multiplicative weights have served as a cornerstone for models of physiological data and neural systems for decades. Although the model discussed here does not implement detailed physiology of synaptic transmission, its new learning laws operate in a network architecture that suggests how recently discovered synaptic computations such as RSE may help produce new network capabilities such as learning that is fast, stable, and distributed. PMID- 11936966 TI - Mean-field approaches to independent component analysis. AB - We develop mean-field approaches for probabilistic independent component analysis (ICA). The sources are estimated from the mean of their posterior distribution and the mixing matrix (and noise level) is estimated by maximum a posteriori (MAP). The latter requires the computation of (a good approximation to) the correlations between sources. For this purpose, we investigate three increasingly advanced mean-field methods: the variational (also known as naive mean field) approach, linear response corrections, and an adaptive version of the Thouless, Anderson and Palmer (1977) (TAP) mean-field approach, which is due to Opper and Winther (2001). The resulting algorithms are tested on a number of problems. On synthetic data, the advanced mean-field approaches are able to recover the correct mixing matrix in cases where the variational meanfield theory fails. For handwritten digits, sparse encoding is achieved using nonnegative source and mixing priors. For speech, the mean-field method is able to separate in the underdetermined (overcomplete) case of two sensors and three sources. One major advantage of the proposed method is its generality and algorithmic simplicity. Finally, we point out several possible extensions of the approaches developed here. PMID- 11936967 TI - Neural networks with local receptive fields and superlinear VC dimension. AB - Local receptive field neurons comprise such well-known and widely used unit types as radial basis function (RBF) neurons and neurons with center-surround receptive field. We study the Vapnik-Chervonenkis (VC) dimension of feedforward neural networks with one hidden layer of these units. For several variants of local receptive field neurons, we show that the VC dimension of these networks is superlinear. In particular, we establish the bound Omega(W log k) for any reasonably sized network with W parameters and k hidden nodes. This bound is shown to hold for discrete center-surround receptive field neurons, which are physiologically relevant models of cells in the mammalian visual system, for neurons computing a difference of gaussians, which are popular in computational vision, and for standard RBF neurons, a major alternative to sigmoidal neurons in artificial neural networks. The result for RBF neural networks is of particular interest since it answers a question that has been open for several years. The results also give rise to lower bounds for networks with fixed input dimension. Regarding constants, all bounds are larger than those known thus far for similar architectures with sigmoidal neurons. The superlinear lower bounds contrast with linear upper bounds for single local receptive field neurons also derived here. PMID- 11936968 TI - Gene transfer to the nonhuman primate retina with recombinant feline immunodeficiency virus vectors. AB - We hypothesize that recombinant feline immunodeficiency viral (rFIV) vectors may be useful for gene transfer to the nonhuman primate retina. We performed vitrectomies and subretinal injections in the right eyes of 11 cynomolgus monkeys. Vesicular stomatitis virus glycoprotein-pseudotyped rFIV that expressed the Escherichia coli beta-galactosidase gene was injected into eight eyes. Sham vehicle or lactose buffer injections were also performed in two of these eight study eyes. rFIV pseudotyped with an amphotropic envelope was used in two eyes, and in one animal injections of lactose buffer only were given. After surgery the animals were clinically evaluated by retinal photography and electroretinography. beta-Galactosidase expression was evaluated, at a final end point, in histological sections. We found photoreceptor and Muller cells to have the greatest transgene expression. Focal inflammatory responses localized to the injection site were seen histologically in all eyes. No difference in transduction efficiency was seen between injections near the macula and more peripheral injections. Visual function as assessed by electroretinography was not significantly affected by vector or vehicle injections. We conclude that rFIV vectors administered beneath the retina can transduce a variety of retinal cells in the nonhuman primate retina. rFIV vectors have therapeutic potential and could be exploited to develop gene therapy for the human eye. PMID- 11936969 TI - Spatial and temporal control of transgene expression through ultrasound-mediated induction of the heat shock protein 70B promoter in vivo. AB - Adenoviral vectors have been constructed that express the transgenes luciferase (Adeno-HSP-Luc) or Fas ligand (Adeno-HSP-FasL) under the control of the heat shock protein 70B (hsp70B) promoter. Cultures infected with Adeno-HSP-Luc transiently expressed high levels of luciferase after heat shock. When cultures infected with Adeno-HSP-FasL were maintained at 37 degrees C, no transgene expression was observed, but when cultures were exposed to heat stress, transgene expression resulted in apoptotic cell death. In vivo, transgene expression was induced by ultrasound-mediated heating of adenovirus-infected tissue. In mice or rats injected with the Adeno-HSP-Luc construct, high levels of localized expression of luciferase activity were observed in regions subjected to ultrasound-mediated irradiation. Adeno-HSP-FasL was administered systemically to mice via the tail vein to evaluate safety. Animals receiving Adeno-HSP-FasL in the absence of ultrasound treatment did not display liver toxicity, whereas animals receiving ultrasound treatment to induce the expression of Fas ligand from the hsp70B promoter had significant increases in serum levels of liver enzymes. These data demonstrate that combining the inducible hsp70B promoter with ultrasound induction allows safe local expression of cytotoxic genes with possible therapeutic utility. PMID- 11936970 TI - Dystrophin gene repair in mdx muscle precursor cells in vitro and in vivo mediated by RNA-DNA chimeric oligonucleotides. AB - Point mutations in the dystrophin gene cause dystrophin deficiency and muscular dystrophy in the mdx mouse and a subset of patients with Duchenne muscular dystrophy. As an approach to gene therapy for muscular dystrophies due to point mutations, we have studied the ability of RNA-DNA chimeric oligonucleotides (chimeraplasts) to induce repair of the dystrophin gene in mdx mice. We have previously demonstrated that targeting chimeraplasts can repair the exon 23 point mutation in differentiated myofibers in vivo after intramuscular injection. For long-term benefit to patients with muscular dystrophy, any gene therapy technology must target not only differentiated myofibers but also undifferentiated muscle precursor cells that are involved in ongoing muscle repair. The focus of the current studies was to test whether chimeraplasts could repair the dystrophin mutation in mdx muscle precursor cells. Initial studies were done by transfecting a targeting chimeraplast into mdx myoblasts in vitro. Gene repair was demonstrated at the DNA, RNA, and protein levels in these cells, whereas treatment of the cells with a control chimeraplast resulted in no gene correction. After differentiation of mdx cells that had been treated with a targeting chimeraplast, immunoblot analysis demonstrated full-length dystrophin expression. By quantitative analysis of independent cultures, the amount of dystrophin expressed ranged from 2 to 15% of that expressed in wild-type cells, providing a measure of the efficacy of gene conversion in vitro. To extend the assessment to muscle precursor cells in vivo, we injected targeting and control chimeraplasts into muscles of mdx mice. When muscle precursor cells were subsequently derived from muscles injected with a targeting chimeraplast, we found that gene repair had occurred in these cells as well. These results, taken together, further demonstrate that chimeraplast-mediated gene repair may be effective as an approach to gene therapy for muscular dystrophies due to point mutations. PMID- 11936971 TI - Pseudorabies virus-based gene delivery to rat embryonic spinal cord grafts. AB - The construction and application of recombinant pseudorabies viruses (PrVs) for the delivery of beta-galactosidase and/or green fluorescent protein (GFP) genes to rat embryonic spinal cord cells are reported here. These viruses were specifically designed to infect embryonic spinal cord neurons, which can be grafted into a lesioned spinal cord in order to restore the lost functions of the host cord. The recombinant viruses were constructed in two steps. The small subunit of the ribonucleotide reductase (RR) gene was first abolished by a frameshift mutation and an expression cassette containing the lacZ gene alone or together with the GFP gene was then inserted in place of the early protein 0 (EP0) gene of PrV. The reporter gene cassettes were positioned downstream from the PrV latency-associated promoter. Using an ex vivo system, we infected embryonic spinal cord explants with these viruses and found that neither vRREP0lac nor vRREP0lacgfp exerted any cytotoxic effect at all. It was also revealed that these viruses infect embryonic cells with high efficiency, and that infected neurons grafted into the spinal cord express the inserted reporter genes for periods of up to 12 weeks. This system offers a new approach for foreign gene transfer to neurons grafted into the CNS. PMID- 11936973 TI - Novel integrating adenoviral/retroviral hybrid vector for gene therapy. AB - A hybrid adenoviral vector system was designed to incorporate an excisable retroviral cassette that can be stably integrated into the host cell genome. The vector contains the terminal sequences of two Moloney murine leukemia virus retroviral long terminal repeats (LTRs), fused to form a junction fragment, and is flanked by two loxP recognition sequences. Cre recombinase-directed excision liberates a circular, double-stranded DNA molecule containing the LTR junction fragment. Despite the natural intermediate for retroviral integrase being a linear DNA molecule, we show that, in the presence of Cre and retroviral Gag and Pol, the excised circle can be integrated into the target cell genome through both specific integrase (Int)-directed mechanisms and by a random integration process. The loxP cassette, carrying in addition a selectable marker gene, was incorporated into the E1-deleted region of an adenoviral vector. Infection of cells expressing Cre, Gag, and Pol generated clones that survived long term in drug selection (>3 months). Int-mediated integration was demonstrated in seven of nine clones by sequencing of the integration sites. In addition, the introduction of the loxP cassette into 293 cells coexpressing Cre and Int alone in the absence of other Gag and Pol proteins was sufficient to catalyze the integration mechanism. These experiments demonstrate that it is possible to generate high titer adenovirus-mediated delivery of a C-type retroviral provirus that can subsequently undergo retroviral Int-mediated integration into dividing and nondividing cells. PMID- 11936972 TI - Intratumoral delivery of adenovirus-mediated interleukin-12 gene in mice with metastatic cancer in the liver. AB - Clinical trials of recombinant human interleukin-12 (rhIL-12) delivered by intravenous administration have shown dose-limiting toxicities with limited tumor responses at the doses tested. We have previously reported that intratumoral injection of an adenovirus vector expressing murine interleukin-12 (Adv.RSV-mIL 12) was effective in inducing antitumor immune responses, tumor regression, and long-term survival in mice with established metastatic cancer in the liver. We now report additional studies in the same murine tumor model to assess the safety of intratumoral Adv.RSV-mIL-12 injection. At vector doses that were previously shown to be therapeutically effective, no inflammation in the liver or lungs, and no significant elevations in serum creatinine and aminotransferases were seen after vector injection. Serum elevations of IL-12 and interferon-gamma (IFN gamma) were 17- and 19-fold lower than peak levels after intravenous recombinant IL-12 at the maximal tolerated dose in clinical trials. No elevations in serum proinflammatory cytokines (interleukin-6, tumor necrosis factor-alpha) were noted up to 2 weeks after vector injection. No systemic dissemination of the vector was detected on polymerase chain reaction (PCR) assays at therapeutically effective vector doses. At higher supratherapeutic vector doses of Adv.RSV-mIL-12, however, inflammation in the liver and lungs with elevation in serum aminotransferases were seen, but not in controls injected with the equivalent particle number of an empty adenoviral vector. These results support the cautious testing in patients with hepatic metastases of adenovirus mediated IL-12 gene delivery by intratumoral injection. PMID- 11936974 TI - p53 and PTEN/MMAC1/TEP1 gene therapy of human prostate PC-3 carcinoma xenograft, using transferrin-facilitated lipofection gene delivery strategy. AB - We previously reported that supplementation of a cationic liposome with transferrin (Tf) greatly enhanced lipofection efficiency (P.-W. Cheng, Hum. Gene Ther. 1996;7:275-282). In this study, we examined the efficacy of p53 and PTEN tumor suppressor gene therapy in a mouse xenograft model of human prostate PC-3 carcinoma cells, using a vector consisting of dimyristoyloxypropyl-3 dimethylhydroxyethyl ammonium bromide (DMRIE)-cholesterol (DC) and Tf. When the volume of the tumors grown subcutaneously in athymic nude mice reached 50-60 mm(3), three intratumoral injections of the following four formulations were performed during week 1 and then during week 3: (1) saline, (2) DC + Tf + pCMVlacZ, (3) DC + Tf + pCMVPTEN, and (4) DC + Tf + pCMVp53 (standard formulation). There was no significant difference in tumor volume and survival between group 1 and group 2 animals. As compared with group 1 controls, group 3 animals had slower tumor growth during the first 3 weeks but thereafter their tumor growth rate was similar to that of the controls. By day 2 posttreatment, group 4 animals had significantly lower tumor volume relative to initial tumor volume as well as controls at the comparable time point. Also, animals treated with p53 survived longer. Treatment with DC, Tf, pCMVp53, DC + pCMVp53, or Tf + pCMVp53 had no effect on tumor volume or survival. Expression of p53 protein and apoptosis were detected in tumors treated with the standard formulation, thus associating p53 protein expression and apoptosis with efficacy. However, p53 protein was expressed in only a fraction of the tumor cells, suggesting a role for bystander effects in the efficacy of p53 gene therapy. We conclude that intratumoral gene delivery by a nonviral vector consisting of a cationic liposome and Tf can achieve efficacious p53 gene therapy of prostate cancer. PMID- 11936990 TI - Long-term effects of inhaled corticosteroids in asthmatic children. PMID- 11936991 TI - Clinical Trials Report. PMID- 11936992 TI - Clinical Trials Report. PMID- 11936993 TI - Clinical Trials Report. PMID- 11936994 TI - Clinical Trials Report. PMID- 11936995 TI - Clinical Trials Report. PMID- 11936996 TI - Clinical Trials Report. PMID- 11936997 TI - Health-related quality of life in high-grade glioma patients. PMID- 11936998 TI - Primitive neuroectodermal tumors/medulloblastoma. AB - The therapy for medulloblastoma/primitive neuroectodermal tumors of the central nervous system is surgery, followed by combination chemo-radiotherapy. The radiation field is the entire craniospinal axis, which is only avoided when treating infants. The treatment is, therefore, lengthy and toxic. Less aggressive therapy is given to patients who clinically appear to have less evidence of disease. Intensive basic research has begun to identify genetic factors of the disease, but these remain far from clinical application. PMID- 11936999 TI - Primary central nervous system lymphoma: a review. AB - Primary central nervous system lymphoma (PCNSL) is an aggressive non-Hodgkin's lymphoma associated with a poor prognosis without treatment. The incidence in immunocompetent patients appears to be increasing. Patients present with nonspecific neurologic symptoms or visual symptoms and contrast-enhancing lesions that abut a cerebrospinal fluid (CSF) space. A subset of patients have positive CSF cytology or ocular involvement. Tissue diagnosis is accomplished by stereotactic biopsy, CSF cytology, or vitreous aspirate. Corticosteroids may obscure the results of tissue specimens and are best avoided prior to tissue diagnosis. PCNSL has emerged as a treatment- sensitive tumor responsive to corticosteroids, radiotherapy, and methotrexate-based chemotherapy. The most effective treatment program has not yet been identified, but it is clear that regimens containing high-dose methotrexate improve survival over radiotherapy alone. Because combined chemo-radiotherapy has been associated with late neurologic toxicities, especially in patients over the age of 60 years, chemotherapy without radiotherapy is being explored further. Significant progress in the treatment of PCNSL has been made in the past 10 years. Further progress will depend heavily on improved understanding of lymphoma biology, and future trials need to focus on improving survival rates while avoiding late neurologic toxicity. PMID- 11937000 TI - Chemotherapeutic dose intensification for treatment of malignant brain tumors: recent developments and future directions. AB - Despite a large amount of research on malignant brain tumors over the past 70 years, the prognosis for most tumor types is poor. One current focus of research is increasing dose intensity of chemotherapeutic agents. Various ways to increase dose intensity include high-dose chemotherapy followed by stem cell rescue (eg, bone marrow transplant), blood-brain barrier disruption or use of RMP7 to increase transvascular drug delivery, local delivery of chemotherapeutic agents (convection enhancement or clysis, antibody conjugates, and biodegradable polymers), chemoprotective agents, and tumor sensitizers. Improved identification of patients likely to respond to a given regimen may also increase the effectiveness of chemotherapy. We also discuss approaches to improve the design of nonrandomized trials by identifying and controlling potential confounding variables. This will improve the quality of individual studies and perhaps the comparability across studies. PMID- 11937001 TI - Neoplastic meningitis. AB - Neoplastic meningitis (NM) is a common problem in neuro-oncology, occurring in approximately 5% of all patients with cancer. Notwithstanding frequent focal signs and symptoms, NM is a disease that affects the entire neuraxis; therefore, staging and treatment need to encompass all cerebrospinal fluid (CSF) compartments. Central nervous system (CNS) staging of NM includes contrast enhanced cranial computerized tomography or magnetic resonance imaging, contrast enhanced spine magnetic resonance imaging or computerized tomographic myelography, and radionuclide CSF flow study. Treatment of NM involves involved field radiotherapy of bulky or symptomatic disease sites and intra-CSF drug therapy. The inclusion of concomitant systemic therapy may benefit patients with NM and may obviate the need for intra-CSF chemotherapy. At present, intra-CSF drug therapy is confined to three chemotherapeutic agents (ie, methotrexate, cytosine arabinoside, and thio-triethylene thiophosphoramide) administered by a variety of schedules either by intralumbar or intraventricular drug delivery. Although treatment of NM is palliative with an expected median patient survival of 2 to 6 months, it often affords stabilization and protection from further neurologic deterioration in patients with NM. PMID- 11937004 TI - Treatment of patients with multiple sclerosis with anti-herpes virus therapy. PMID- 11937002 TI - Pituitary tumor diagnosis and treatment. AB - Over the past few years, significant contributions have been made to the understanding, diagnosis, and treatment of pituitary tumors. This article reviews recent advances in the areas of biology, diagnostic imaging, medical diagnosis and treatment, surgical results and technique, and adjuvant therapy in the form of radiotherapy and radiosurgery. Of particular note are the roles of endoscopy, intraoperative magnetic resonance imaging, radiosurgery, and radiation for nonfunction tumors, the diagnosis of Cushing's disease, the management of "incidentalomas," and new medication therapies. PMID- 11937003 TI - Mechanisms of apoptosis in central nervous system tumors: application to theory. AB - Apoptosis is a key concept for the successful therapy of brain tumors. This review focuses on the mechanisms of apoptosis occurring spontaneously in malignant gliomas, discusses the different methods employed to assess apoptosis in vivo and in vitro, and considers the value of quantifying apoptosis in surgical biopsies for diagnosis and prognosis. Further, novel strategies to induce apoptosis in human malignant glioma cells are reviewed, including experimental therapy with death ligands, methods for sensitizing glioma cells to the induction of apoptosis, p53 gene transfer, and approaches to target the expression of therapeutic genes selectively to tumor cells. PMID- 11937005 TI - Genetic analysis of multiple sclerosis. AB - The increased recurrence risk within families indicates a role for genetic factors in the etiology of multiple sclerosis. Genes may influence susceptibility to the development of multiple sclerosis and the subsequent course of the disease. To date, associations have only been demonstrated consistently with class II major histocompatibility complex (MHC) alleles. The relatively low yield from additional candidate gene studies is only modestly advanced by several whole genome linkage analyses, and by the first in a series of planned whole-genome linkage disequilibrium screens for allelic associations. The aims of linkage and association are to narrow the search for chromosomal regions encoding genes for multiple sclerosis and, with information from the human gene project, suggest new positional candidates. In time, it is expected that these genes will include some that confer susceptibility to the general process of autoimmunity, others that are specific for multiple sclerosis in all populations, some that act only in defined ethic groups, and those that determine particular phenotypes or shape the clinical course. These genetic analyses are predicated on the assumption that multiple sclerosis is one disease; a major part of future studies will be to resolve the question of disease heterogeneity in multiple sclerosis. When eventually in place, the potential of this genetic knowledge for improved understanding of the pathogenesis of multiple sclerosis and designing novel treatments is considerable. PMID- 11937008 TI - Combination chemotherapy for ovarian cancer--lessons not yet learned? PMID- 11937006 TI - Clinical trials and clinical practice in multiple sclerosis: conventional and emerging magnetic resonance imaging technologies. AB - Conventional magnetic resonance imaging (cMRI) is widely used for diagnosing multiple sclerosis (MS) and as a paraclinical tool to monitor disease activity and evolution in natural history studies and clinical trials. However, the correlation between cMRI and clinical findings is far from strict, and such a discrepancy is even more evident when moving from the setting of large-scale studies to the management of individual patients. Among the reasons for this "clinical-MRI paradox" is the limited specificity of cMRI to the heterogeneous pathologic substrates of MS and its inability to quantify the extent of damage in the normal-appearing tissues. Modern quantitative MR techniques have the potential to overcome some of the limitations of cMRI. Although the application of modern MR techniques is changing dramatically our understanding of how MS causes irreversible disability, their use for clinical trial monitoring and clinical practice is still very limited. Whereas there is increasing perception that modern quantitative MR techniques should be more extensively employed in clinical trials to advance the understanding of MS and derive innovative information, their use in clinical practice should still be regarded as premature. PMID- 11937007 TI - Gender issues and multiple sclerosis. AB - Gender-related issues in multiple sclerosis include the important and widely accepted clinical observations that men are less susceptible to the disease than women and also that disease activity in multiple sclerosis is decreased during late pregnancy. This article reviews mechanisms underlying each of these clinical observations and discusses the role of sex hormones in each. Specifically, the protective role of testosterone in young men and the protective role of the pregnancy hormone estriol in pregnant women are discussed. Rationale for novel therapies in multiple sclerosis based on the protective roles of these sex hormones is presented. PMID- 11937009 TI - Recent developments in the treatment of gastric carcinoma. AB - Surgery remains the mainstay for the curative treatment of gastric carcinoma. However, despite adequate surgery, survival remains poor. The use of adjuvant chemotherapy and radiotherapy has been examined in multiple previous clinical trials without convincing evidence of efficacy. However, recently, a large randomized controlled Intergroup trial, INT 116, demonstrated a survival advantage with chemoradiotherapy following curative surgery for gastric cancer. This review discusses the merits of the Intergroup trial and the ways in which it should affect the treatment of gastric cancer in the United States. INT 116 provides a foundation on which we can build to improve the care of patients with gastric cancer. With the evaluation of potentially better chemotherapeutic agents and the advent of molecularly directed therapy, there is increasing hope for improving the care of patients with gastric carcinoma. PMID- 11937010 TI - Current and future strategies for combined-modality therapy in pancreatic cancer. AB - Treatment of pancreatic cancer remains a challenging task that often requires a multidisciplinary approach to confer optimal response and, ideally, maximize survival. A combination of locoregional approaches such as surgery and radiotherapy, along with systemic therapies for eradication of micrometastases, should be considered both for patients who are operative candidates and for those with locally advanced, unresectable disease. How best to combine these modalities in terms of schedule, timing, and choice of agents is a question that continues to be actively investigated. Some of these data are equivocal or conflicting; thus standards of care for combined-modality treatment have not been uniformly accepted to date. This article provides an overview of combined-modality therapy, focusing on the major studies that have guided our current approach to the treatment of pancreatic cancer and examining new strategies that are likely to improve outcomes and survival for patients in the future. PMID- 11937011 TI - New developments in the treatment of esophageal cancer. AB - Esophageal cancer is a rare but highly virulent malignancy in the United States and Western Europe, and adenocarcinoma of the esophagus has had the most rapid rate of increase of any solid tumor malignancy. Combined chemoradiotherapy is the standard of care in the nonsurgical management of esophageal cancer. Trials of preoperative chemotherapy followed by surgery have not shown a consistent benefit. Preoperative chemoradiotherapy followed by surgery continues to be actively studied in the surgical management of locally advanced esophageal cancer. Pathologic complete responses are seen in 20% to 40% of patients, with 5 year survival achieved in 30% to 35%. Newer agents, such as the taxanes and irinotecan, have been evaluated in combined chemoradiotherapy trials. These trials have shown promising antitumor activity and therapy tolerance, depending on the dose and schedule of therapy administered. Increasing the dose of radiotherapy, or adding a brachytherapy boost to chemoradiotherapy, has not improved the outcome of treatment in clinical trials. The advent of newer targeted therapies, including agents directed against growth factor receptor pathways, tumor angiogenesis, and tumor invasion and metastasis, is leading to a new generation of clinical trials combining these agents with conventional cytotoxic chemotherapy and radiation. PMID- 11937013 TI - Adenosine triphosphate for cancer cachexia. PMID- 11937012 TI - Adjuvant and preoperative chemotherapy for gastric cancer. AB - Gastric cancer is the second most frequently diagnosed malignancy worldwide, and the risk of relapse remains high in the majority of patients undergoing resection. Attempts to reduce this risk and prolong survival have led to numerous adjuvant chemotherapy trials that either had no benefit for patients or occasionally had controversial results. The recently reported Intergroup 0116 trial shows conclusive evidence of survival benefit for patients treated with postoperative chemoradiotherapy. In this Intergroup trial, which involves over 600 patients, a regimen of postoperative chemotherapy plus chemoradiotherapy was shown to prolong overall and disease-free survival in gastric cancer patients with stage IB through IV disease following a curative (R0) resection. This approach should be considered the standard of care in patients with gastric cancer who have undergone curative resection. Preoperative chemotherapy shows promise in downstaging tumors and increasing the rate of curative resection, but randomized trials are needed to assess survival benefits. Efforts to combine existing treatment modalities and new agents with novel mechanisms of action hold promise for the future. PMID- 11937014 TI - Surgical palliation in advanced disease: recent developments. AB - Despite many valuable technical innovations for the relief of suffering in advanced disease over the past few years, only recently have surgical oncologists attempted to more clearly define palliation. Previous definitions have been misleading, creating confusion about the merits of surgery in many situations and difficulty in posing questions for future prospective clinical trials. This report outlines recent progress in identifying and refining a philosophy of palliative surgery that would align it with the consensus of nonsurgical opinion summarized by the 1990 World Health Organization definition of palliative care and the emerging consensus among the medical specialties in the United States concerning principles of care at the end of life. Selected controversies and recent innovations, as well as guidelines for palliative surgery, are discussed. PMID- 11937015 TI - Practical guide to palliative sedation. AB - Terminally ill patients want assurance that their symptoms will be controlled as death approaches. Most patients can have a peaceful death with standard palliative care. Some patients approaching death, however, have refractory symptoms such as pain, dyspnea, nausea, and agitated delirium. Palliative sedation (PS), the use of medications to induce sedation in order to control refractory symptoms near death, is a therapeutic option for these patients. The reported frequency of PS use varies greatly, ranging from 5% to 52% of the terminally ill. One concern with PS is its effect on survival. Data suggest that PS does not lead to immediate death, with the median time to death after initiating PS being greater than 1 to 5 days. A number of medications have been used for PS, but midazolam is most commonly reported. PS is distinct from euthanasia because the intent of PS is relief from suffering without death as a required outcome. PMID- 11937016 TI - The role of C-reactive protein as a prognostic indicator in advanced cancer. AB - C-reactive protein (CRP) is a nonspecific but sensitive marker of inflammation. Interleukin-6 (IL-6), IL-1, and tumor necrosis factor alpha induce the synthesis of CRP in hepatocytes. Increased CRP level is considered to be an important risk factor for atherosclerosis, myocardial infarction, peripheral vascular disease, and ischemic stroke. It is positively correlated with weight loss, anorexia cachexia syndrome, extent of disease, and recurrence in advanced cancer. Its role as a predictor of survival has been shown in multiple myeloma, melanoma, lymphoma, ovarian, renal, pancreatic, and gastrointestinal tumors. Measurement of CRP is simple, cheap, and routine and provides valuable information in palliative care. PMID- 11937017 TI - Symptom assessment in cancer patients. AB - Patients with cancer suffer from a multitude of symptoms. Effective treatment strategies are available for a number of the common symptoms. A major reason for inadequate symptom relief is lack of effective symptom assessment. Accurate assessment of symptoms is necessary before any treatment can be undertaken. Although there are many complexities involved in the assessment of symptoms, simple, effective tools are available for identification and scoring of symptoms in clinical practice. This review describes recent advances in the study of symptom assessment in cancer patients. Several common symptoms are presented individually, followed by an overview of efforts addressing the assessment of multiple symptoms. Also, new tools in the evaluation of symptoms, including computer-based questionnaires, are discussed. PMID- 11937019 TI - Spontaneous mutations in KNOX genes give rise to a novel floral structure in Antirrhinum. AB - BACKGROUND: Petal spurs-tubular outgrowths that collect nectar-are considered key innovations because of their ability to change pollinator specificity and so cause reproductive isolation and speciation. Spurs have arisen frequently and rapidly in many taxa. To test their potential origins, we isolated spontaneous dominant mutations at two loci, HIRZ and INA, that cause novel outgrowths from Antirrhinum petals, resembling the petal spurs of closely related genera. RESULTS: HIRZ and INA were isolated and shown to encode similar KNOX homeodomain proteins that are normally expressed only in apical meristems and are likely to act redundantly. Both dominant mutations were caused by transposon insertions in noncoding regions that caused ectopic expression of functional transcripts, either in petals or in all lateral organs with more pleiotropic effects. Formation of a spur-like outgrowth, which resembled an ectopic petal tube, was dependent both on KNOX gene expression and dorsiventral asymmetry of the flower. CONCLUSIONS: These mutations provide an example of how petal spurs might evolve rapidly due to changes in regulatory gene expression. PMID- 11937018 TI - New drugs for the anorexia-cachexia syndrome. AB - Anorexia and cachexia accompany advancing cancer to a greater extent than any other symptom. Cachexia alone causes 22% of cancer deaths. The pathophysiology of cachexia is distinctly different from that of starvation. Resting energy expenditures are elevated, and abnormal intermediary metabolism, proteolysis, and lipolysis occur independently of caloric intake. A facilatative interaction between catecholamines, prostaglandins, and inflammatory cytokines is responsible for cachexia. Successful treatment requires reduction of energy expenditures, reversal of anorexia, and correction of abnormal intermediary metabolism, lipolysis, and proteolysis. Multiple appetite stimulants can be used in combination. Several new potentially useful biologic agents have been tested in animal tumor models. Several of the anticachectic agents have demonstrated in vivo or in vitro antitumor activity. The biologic and clinical activity of each drug is reviewed herein, and potentially useful combinations are listed. PMID- 11937020 TI - Transfer of scrapie prion infectivity by cell contact in culture. AB - BACKGROUND: When a cell is infected with scrapie prions, newly synthesized molecules of the prion protein PrP(C) are expressed at the cell surface and may subsequently be converted to the abnormal form PrP(Sc). In an experimental scrapie infection of an animal, the initial innoculum of PrP(Sc) is cleared relatively rapidly, and the subsequent propagation of the infection depends on the ability of infected cells to convert uninfected target cells to stable production of PrP(Sc). The mechanism of such cell-based infection is not understood. RESULTS: We have established a system in dissociated cell culture in which scrapie-infected mouse SMB cells are able to stably convert genetically marked target cells by coculture. After coculture and rigorous removal of SMB cells, the target cells express PrP(Sc) and also incorporate [35S]methionine into PrP(Sc). The extent of conversion was sensitive to the ratio of the two cell types, and conversion by live SMB required 2500-fold less PrP(Sc) than conversion by a cell-free prion preparation. The conversion activity of SMB cells is not detectable in conditioned medium and apparently depends on close proximity or contact, as evidenced by culturing the SMB and target cells on neighboring but separate surfaces. SMB cells were killed by fixation in aldehydes, followed by washing, and were found to retain significant activity at conversion of target cells. CONCLUSIONS: Cell-mediated infection of target cells in this culture system is effective and requires significantly less PrP(Sc) than infection by a prion preparation. Several lines of evidence indicate that it depends on cell contact, in particular, the activity of aldehyde-fixed infected cells. PMID- 11937021 TI - Recruitment of scribble to the synaptic scaffolding complex requires GUK-holder, a novel DLG binding protein. AB - BACKGROUND: Membrane-associated guanylate kinases (MAGUKs), such as Discs-Large (DLG), play critical roles in synapse maturation by regulating the assembly of synaptic multiprotein complexes. Previous studies have revealed a genetic interaction between DLG and another PDZ scaffolding protein, SCRIBBLE (SCRIB), during the establishment of cell polarity in developing epithelia. A possible interaction between DLG and SCRIB at synaptic junctions has not yet been addressed. Likewise, the biochemical nature of this interaction remains elusive, raising questions regarding the mechanisms by which the actions of both proteins are coordinated. RESULTS: Here we report the isolation of a new DLG-interacting protein, GUK-holder, that interacts with the GUK domain of DLG and which is dynamically expressed during synaptic bouton budding. We also show that at Drosophila synapses DLG colocalizes with SCRIB and that this colocalization is likely to be mediated by direct interactions between GUKH and the PDZ2 domain of SCRIB. We show that DLG, GUKH, and SCRIB form a tripartite complex at synapses, in which DLG and GUKH are required for the proper synaptic localization of SCRIB. CONCLUSIONS: Our results provide a mechanism by which developmentally important PDZ-mediated complexes are associated at the synapse. PMID- 11937022 TI - Circadian cycling of the mouse liver transcriptome, as revealed by cDNA microarray, is driven by the suprachiasmatic nucleus. AB - BACKGROUND: Genes encoding the circadian pacemaker in the hypothalamic suprachiasmatic nuclei (SCN) of mammals have recently been identified, but the molecular basis of circadian timing in peripheral tissue is not well understood. We used a custom-made cDNA microarray to identify mouse liver transcripts that show circadian cycles of abundance under constant conditions. RESULTS: Using two independent tissue sampling and hybridization regimes, we show that approximately 9% of the 2122 genes studied show robust circadian cycling in the liver. These transcripts were categorized by their phase of abundance, defining clusters of day- and night-related genes, and also by the function of their products. Circadian regulation of genes was tissue specific, insofar as novel rhythmic liver genes were not necessarily rhythmic in the brain, even when expressed in the SCN. The rhythmic transcriptome in the periphery is, nevertheless, dependent on the SCN because surgical ablation of the SCN severely dampened or destroyed completely the cyclical expression of both canonical circadian genes and novel genes identified by microarray analysis. CONCLUSIONS: Temporally complex, circadian programming of the transcriptome in a peripheral organ is imposed across a wide range of core cellular functions and is dependent on an interaction between intrinsic, tissue-specific factors and extrinsic regulation by the SCN central pacemaker. PMID- 11937023 TI - Circadian programs of transcriptional activation, signaling, and protein turnover revealed by microarray analysis of mammalian cells. AB - Many aspects of physiology and behavior are temporally organized into daily 24 hr rhythms, driven by an endogenous circadian clock. Studies in eukaryotes have identified a network of interacting genes forming interlocked autoregulatory feedback loops which underlie overt circadian organization in single cells. While in mammals the master oscillator resides in the suprachiasmatic nuclei of the hypothalamus, semiautonomous circadian oscillators also exist in peripheral tissues and in immortalized fibroblasts, where rhythmicity is induced following a serum shock. We used this model system in combination with high-density cDNA microarrays to examine the magnitude and quality of clock control of gene expression in mammalian cells. Supported by application of novel bioinformatics tools, we find approximately 2% of genes, including expected canonical clock genes, to show consistent rhythmic circadian expression across five independent experiments. Rhythmicity in most of these genes is novel, and they fall into diverse functional groups, highlighted by a predominance of transcription factors, ubiquitin-associated factors, proteasome components, and Ras/MAPK signaling pathway components. When grouped according to phase, 68% of the genes were found to peak during estimated subjective day, 32% during estimated subjective night, with a tendency to peak at a phase corresponding to anticipation of dawn or dusk. PMID- 11937024 TI - UUCAC- and vera-dependent localization of VegT RNA in Xenopus oocytes. AB - Localized mRNAs are directed to their destinations by "localization elements" (LEs) in their 3'UTRs. LEs harbor multiple, functionally redundant localization "signals." These signals are poorly defined, hence it is unclear whether the signals-and their cognate factors-are unique to each RNA or employed generally. Five "E2s" (UUCACs) in the 366 nt Vg1 LE (VgLE) direct this transcript to the vegetal pole of Xenopus oocytes via the binding of a protein-Vera/Vg1RBP/ZBP. Here we show that a different vegetal RNA, VegT, employs the same signal and factor. Five E2s within a 440 nt subregion (VegT440) of the VegT 3'UTR predict its LE and are both necessary and sufficient (in the context of antisense VegT440) for directing localization. The E2s in VegT440 and VgLE function similarly to recruit Vera protein: (1) in both contexts, E2 nt substitutions partially (UU to AC) or completely (CA to UG) inhibit localization in accordance with the sequence selectivity of Vera protein for E2s; (2) VegT440 and VgLE crosscompete, in an E2-dependent manner, for localization and Vera binding; (3) injection of anti-Vera antibody into oocytes inhibits localization of both injected transcripts. These findings imply that general localization signals traffic diverse RNAs. PMID- 11937025 TI - Ena/VASP proteins regulate cortical neuronal positioning. AB - Development of the multilayered cerebral cortex involves extensive regulated migration of neurons arising from the deeper germinative layers of the mammalian brain. The anatomy and formation of the cortical layers has been well characterized; however, the underlying molecular mechanisms that control the migration and the final positioning of neurons within the cortex remain poorly understood. Here, we report evidence for a key role of Ena/VASP proteins, a protein family implicated in the spatial control of actin assembly and previously shown to negatively regulate fibroblast cell speeds, in cortical development. Ena/VASP proteins are highly expressed in the developing cortical plate in cells bordering Reelin-expressing Cajal-Retzius cells and in the intermediate zone through which newly born cells migrate. Inhibition of Ena/VASP function through retroviral injections in utero led to aberrant placement of early-born pyramidal neurons in the superficial layers of both the embryonic and the postnatal cortex in a cell-autonomous fashion. The abnormally placed pyramidal neurons exhibited grossly normal morphology and polarity. Our results are consistent with a model in which Ena/VASP proteins function in vivo to control the position of neurons in the mouse neocortex. PMID- 11937026 TI - A new internal mode in F-actin helps explain the remarkable evolutionary conservation of actin's sequence and structure. AB - Actin is one of the most highly conserved eukaryotic proteins. There are no amino acid changes between the chicken and human skeletal muscle isoforms, and the most dissimilar actins still share more than 85% sequence identity [1]. We suggest that large discrete internal modes of freedom within the actin filament may account for a significant component of this conservation, since each subunit must make multiple specific interactions with neighboring subunits. In support of this, we find that the same state of tilt of the actin subunit exists in both yeast and vertebrate striated muscle actin, and that in both the two domains undergo a "propeller rotation." A similar movement of domains has also been seen in hexokinase, Hsc70, and Arp2/3, all structural homologs of actin, suggesting that such an interdomain hinge motion is common to proteins in this superfamily. Subunit-subunit interactions within the actin filament involve sequence insertions that are not present in MreB, a bacterial homolog of actin. Remarkably, we find that in the tilted state actin subunits make new contacts with neighboring subunits that also involve these inserts, suggesting a key role for these elements in F-actin polymorphism. PMID- 11937027 TI - Notch activation of yan expression is antagonized by RTK/pointed signaling in the Drosophila eye. AB - Receptor tyrosine kinase (RTK) signaling plays an instructive role in cell fate decisions, whereas Notch signaling is often involved in restricting cellular competence for differentiation. Genetic interactions between these two evolutionarily conserved pathways have been extensively documented. The underlying molecular mechanisms, however, are not well understood. Here, we show that Yan, an Ets transcriptional repressor that blocks cellular potential for specification and differentiation, is a target of Notch signaling during Drosophila eye development. The Suppressor of Hairless (Su[H]) protein of the Notch pathway is required for activating yan expression, and Su(H) binds directly to an eye-specific yan enhancer in vitro. In contrast, yan expression is repressed by Pointed (Pnt), which is a key component of the RTK pathway. Pnt binds specifically to the yan enhancer and competes with Su(H) for DNA binding. This competition illustrates a potential mechanism for RTK and Notch signals to oppose each other. Thus, yan serves as a common target of Notch/Su(H) and RTK/Pointed signaling pathways during cell fate specification. PMID- 11937028 TI - Type I PIPkinases interact with and are regulated by the retinoblastoma susceptibility gene product-pRB. AB - Inositide signaling at the plasma membrane has been implicated in the regulation of numerous cellular processes including cytoskeletal dynamics, vesicle trafficking, and gene transcription. Studies have also shown that a distinct inositide pathway exists in nuclei, where it may regulate nuclear processes such as mRNA export, cell cycle progression, gene transcription, and DNA repair. We previously demonstrated that nuclear PtdIns(4,5)P(2) synthesis is stimulated during progression from G1 through S phase, although mechanistic details of how cell cycle progression impinges on the regulation of nuclear inositides is unknown. In this study, we demonstrate that pRB, which regulates progression of cells from G1 through S phase interacts both in vitro and in vivo with Type I PIPkinases, the enzymes responsible for nuclear PtdIns(4,5)P(2) synthesis. Moreover, this interaction stimulates the activity of Type Ialpha PIPkinase in an in vitro assay. Using murine erythroleukamia (MEL) cells expressing a temperature sensitive mutant of large T antigen (LTA), we demonstrate changes in vivo in nuclear PtdIns(4,5)P(2) levels that are consistent with the ability of LTA to disrupt pRB/Type I interactions. This study, for the first time, provides a potential mechanism for how cell cycle progression could regulate the levels of nuclear inositides. PMID- 11937029 TI - The protein kinase C pathway is required for viability in quiescence in Saccharomyces cerevisiae. AB - Protein kinase C, encoded by PKC1, regulates construction of the cell surface in vegetatively growing yeast cells. Pkc1 in part acts by regulating Mpk1, a MAP kinase. Mutants lacking Bck1, a component of the MAP kinase branch of the pathway, fail to respond normally to nitrogen starvation, which causes entry into quiescence. Given that the Tor1 and Tor2 proteins are key inhibitors of entry into quiescence, the Pkc1 pathway may regulate these proteins. We find that pkc1Delta and mpk1Delta mutants rapidly die by cell lysis upon carbon or nitrogen starvation. The Pkc1 pathway does not regulate the TOR proteins: transcriptional changes dependent on inhibition of the TORs occur normally in pkc1Delta and mpk1Delta mutants when starved for nitrogen; pkc1Delta and mpk1Delta mutants die rapidly upon treatment with rapamycin, an inhibitor of the TORs. We find that Mpk1 is transiently activated by rapamycin treatment via a novel mechanism. Finally, we find that rapamycin treatment or nitrogen starvation induces resistance to the cell wall-digesting enzyme zymolyase by a Pkc1-dependent mechanism. Thus, the Pkc1 pathway is not a nutrient sensor but acts downstream of TOR inhibition to maintain cell integrity in quiescence. PMID- 11937030 TI - Histone deacetylase 1 inactivation by an adenovirus early gene product. AB - Gam1 is an early gene product of the avian adenovirus CELO and is essential for viral replication. Gam1 has no homology to any known proteins; however, its early expression and nuclear localization suggest that the protein functions to influence transcription in the infected cell. A determinant of eukaryotic gene expression is the acetylation state of chromosomal histones and other nuclear proteins. We find that Gam1 expression increases the level of transcription from a variety of eukaryotic promoters, similar to the effect of treating cells with the histone deacetylase (HDAC) inhibitor trichostatin A (TSA ). We show that Gam1 can effectively inhibit histone deacetylation by HDAC1 and that Gam1 binds to HDAC1 both in vitro and in vivo. A CELO virus lacking Gam1 (CELOdG) is replication defective, but the defect can be overcome by either expressing an interfering HDAC1 mutant or by treating infected cells with TSA. The identification of a viral early gene product having the specific function of binding and inactivating HDAC suggests that deacetylase complexes play an important role in limiting early gene expression from invading viruses. PMID- 11937031 TI - CDK phosphorylation of Drc1 regulates DNA replication in fission yeast. AB - Cyclin-dependent kinases (CDKs) are absolutely required for DNA replication in eukaryotic cells. CDKs are thought to activate one or more replication factors, but the identities of these proteins are unknown. Here we describe fission yeast Drc1, a protein required for DNA replication that is phosphorylated by Cdc2. Drc1 depletion leads to catastrophic mitotic divisions with incompletely replicated DNA, indicating that Drc1 is required for DNA synthesis and S-M replication checkpoint control. Drc1 associates with Cdc2 and is phosphorylated at the onset of S phase when Cdc2 is activated. Mutant Drc1 that lacks CDK phosphorylation sites is nonfunctional and fails to interact with Cut5 replication factor. These data suggest that Cdc2 promotes DNA replication by phosphorylating Drc1 and regulating its association with Cut5. PMID- 11937032 TI - Functional integration of adult-born neurons. AB - Over the past decade, it has become clear that neural stem cells in the adult mammalian brain continuously generate new neurons, predominantly in the hippocampus and olfactory bulb. However, the central issue of whether these new neurons participate in functional synaptic circuitry has yet to be resolved. Here, we use virus-based transsynaptic neuronal tracing and c-Fos mapping of odor induced neuronal activity to demonstrate that neurons generated in the adult functionally integrate into the synaptic circuitry of the brain. PMID- 11937033 TI - Capturing public unease. PMID- 11937035 TI - The COP9 signalosome. PMID- 11937037 TI - The insulin signalling pathway. PMID- 11937036 TI - Pheromone induces programmed cell death in S. cerevisiae. PMID- 11937038 TI - Aging: from radiant youth to an abrupt end. AB - A gene that controls growth rate, radiation resistance and lifespan in the nematode Caenorhabditis elegans has been found to encode a homologue of a yeast telomere maintenance factor, raising the possibility that checkpoint control, telomere maintenance and aging may be linked in unanticipated ways. PMID- 11937039 TI - Nuclear envelope: torn apart at mitosis. AB - The findings of two recent studies suggest a novel mechanism for nuclear envelope breakdown in which cytoplasmic dynein anchored on the outside of the nucleus generates tension, thus triggering tearing of the nuclear envelope. PMID- 11937040 TI - Chromatin remodeling: nucleosomes bulging at the seams. AB - ATP-dependent chromatin remodeling enzymes, such as SWI/SNF, hydrolyze thousands of ATPs to regulate gene expression on chromatin fibers. Recent mechanistic studies suggest that these enzymes generate localized changes in DNA topology that drive formation of multiple, remodeled nucleosomal states. PMID- 11937041 TI - Prion disease: close encounters of the cellular kind. AB - Co-culture of prion-infected cells with uninfected target cells has recently been found to lead to rapid and efficient transmission of infectivity to the target cells by a process that is dependent on direct cell contact. PMID- 11937042 TI - Water transporters: how so fast yet so selective? AB - A high-resolution X-ray structure of an aquaporin has revealed water molecules bound within the transmembrane pore and provided new clues to the mechanisms of rapid water transport and high selectivity in this important class of membrane proteins. PMID- 11937043 TI - Visual ecology: coloured fruit is what the eye sees best. AB - Trichromatic vision may have evolved as an aid to frugivory. This hypothesis is supported by the recent demonstration that the spatial characteristics of pictures containing fruit are particularly well matched to the abilities of the human visual system. PMID- 11937044 TI - Chromatin assembly: the kinetochore connection. AB - An unexpected new role for the chromatin assembly factor CAF-1 and the histone regulating Hir proteins has been discovered in budding yeast. Both protein complexes are required together for building functional kinetochores. PMID- 11937045 TI - Cell signaling: life or death decisions of ras proteins. AB - The discovery of a new downstream target for the Ras GTPases - a Nore1-Mst1 protein complex - reveals a mechanism by which Ras can promote apoptosis, and suggests that the influence of Ras on cell survival or death depends upon the relative level of activation of its various target proteins. PMID- 11937046 TI - DNA damage: air-breaks? AB - Cells deficient in repairing DNA double-strand breaks have an increased level of spontaneous chromosomal aberrations. Modulating the level of molecular oxygen and its reactive metabolites demonstrates that oxygen metabolism is a major source of genomic instability. PMID- 11937047 TI - HIV-1 transcription: activation mediated by acetylation of Tat. AB - Acetylation of HIV-1 Tat stimulates transcriptional elongation by dissociating Tat from TAR, a transactivation response RNA element in nascent HIV-1 transcripts. In the March issue of Molecular Cell, Mujtaba et al. show that the bromodomain of PCAF acetylase specifically binds to acetylated Tat and leads to dissociation of Tat from TAR. PMID- 11937048 TI - A glimpse of the catalytic core of a group II intron. AB - A paper in a recent issue of Science describes the first high-resolution structure of part of the catalytic core of a group II intron that will allow more detailed comparisons between the excision of introns by self-splicing group II introns and by nuclear pre-mRNA introns. PMID- 11937049 TI - Novel sequences propel familiar folds. AB - Recent structure determinations have made new additions to a set of strikingly different sequences that give rise to the same topology. Proteins with a beta propeller fold are characterized by extreme sequence diversity despite the similarity in their three-dimensional structures. Several fold predictions, based in part on sequence repeats thought to match modular beta sheets, have been proved correct. PMID- 11937050 TI - Max Perutz (1914-2002). Great scientist and modest leader. PMID- 11937051 TI - Homo crystallographicus--quo vadis? AB - As macromolecular crystal structures are determined and refined in an increasingly automated fashion, careful assessment of the reliability and quality of the resulting models becomes increasingly important. Here, we analyze various issues related to the reliability and quality of macromolecular crystal structures deposited between 1991 and 2000. We find that the average resolution at which these structures are determined is essentially constant. In line with this observation, the average quality as measured by Ramachandran analysis does not improve as a function of time. On the other hand, an observed decrease of the average discrepancy between free and conventional R values suggests that the fit of model and data is improving. Finally, we present a surprising correlation between the tendency of crystallographers to deposit their experimental data and the free R values of their models. PMID- 11937052 TI - Early structural rearrangements in the photocycle of an integral membrane sensory receptor. AB - Sensory rhodopsins are the primary receptors of vision in animals and phototaxis in microorganisms. Light triggers the rapid isomerization of a buried retinal chromophore, which the protein both accommodates and amplifies into the larger structural rearrangements required for signaling. We trapped an early intermediate of the photocycle of sensory rhodopsin II from Natronobacterium pharaonis (pSRII) in 3D crystals and determined its X-ray structure to 2.3 A resolution. The observed structural rearrangements were localized near the retinal chromophore, with a key water molecule becoming disordered and the retinal's beta-ionone ring undergoing a prominent movement. Comparison with the early structural rearrangements of bacteriorhodopsin illustrates how modifications in the retinal binding pocket of pSRII allow subtle differences in the early relaxation of photoisomerized retinal. PMID- 11937053 TI - The structure of Neurospora crassa 3-carboxy-cis,cis-muconate lactonizing enzyme, a beta propeller cycloisomerase. AB - Muconate lactonizing enzymes (MLEs) convert cis,cis-muconates to muconolactones in microbes as part of the beta-ketoadipate pathway; some also dehalogenate muconate derivatives of xenobiotic haloaromatics. There are three different MLE classes unrelated by evolution. We present the X-ray structure of a eukaryotic MLE, Neurospora crassa 3-carboxy-cis,cis-muconate lactonizing enzyme (NcCMLE) at 2.5 A resolution, with a seven-bladed beta propeller fold. It is related neither to bacterial MLEs nor to other beta propeller enzymes, but is structurally similar to the G protein beta subunit. It reveals a novel metal-independent cycloisomerase motif unlike the bacterial metal cofactor MLEs. Together, the bacterial MLEs and NcCMLE structures comprise a striking structural example of functional convergence in enzymes for 1,2-addition-elimination of carboxylic acids. NcCMLE and bacterial MLEs may enhance the reaction rate differently: the former by electrophilic catalysis and the latter by electrostatic stabilization of the enolate. PMID- 11937054 TI - The structure and mechanism of the type II dehydroquinase from Streptomyces coelicolor. AB - The structure of the type II DHQase from Streptomyces coelicolor has been solved and refined to high resolution in complexes with a number of ligands, including dehydroshikimate and a rationally designed transition state analogue, 2,3-anhydro quinic acid. These structures define the active site of the enzyme and the role of key amino acid residues and provide snap shots of the catalytic cycle. The resolution of the flexible lid domain (residues 21-31) shows that the invariant residues Arg23 and Tyr28 close over the active site cleft. The tyrosine acts as the base in the initial proton abstraction, and evidence is provided that the reaction proceeds via an enol intermediate. The active site of the structure of DHQase in complex with the transition state analog also includes molecules of tartrate and glycerol, which provide a basis for further inhibitor design. PMID- 11937055 TI - Sperm coating mechanism from the 1.8 A crystal structure of PDC-109 phosphorylcholine complex. AB - Bovine seminal plasma PDC-109 binds to sperm surface choline lipids and promotes sperm capacitation by stimulating the efflux of cholesterol and phospholipids. The structure of PDC-109 with bound phosphorylcholine was solved using MAD data of a single platinum site. Its two globular (40 x 50 x 20 A(3)) Fn2 domains are linked and clustered by a short polypeptide. The choline binding sites lie at the same face of the molecule. Phosphorylcholine binds to the Fn2 domains through a cation-pi interaction between the quaternary ammonium group and a core tryptophan, plus hydrogen bonding between hydroxyls of exposed tyrosines and the phosphate group. The structure of the PDC-109-oPC complex provides a structural ground for the sperm membrane-coating mechanism underlying PDC-109-induced capacitation. PMID- 11937056 TI - Grafting segments from the extracellular surface of CCR5 onto a bacteriorhodopsin transmembrane scaffold confers HIV-1 coreceptor activity. AB - Components from the extracellular surface of CCR5 interact with certain macrophage-tropic strains of human immunodeficiency virus type 1 (HIV-1) to mediate viral fusion and entry. To mimic these viral interacting site(s), the amino-terminal and extracellular loop segments of CCR5 were linked in tandem to form concatenated polypeptides, or grafted onto a seven-transmembrane bacteriorhodopsin scaffold to generate several chimeras. The chimera studies identified specific regions in CCR5 that confer HIV-1 coreceptor function, structural rearrangements in the transmembrane region that may modulate this activity, and a role for the extracellular surface in folding and assembly. Methods developed here may be applicable to the dissection of functional domains from other seven-transmembrane receptors and form a basis for future structural studies. PMID- 11937057 TI - Design, construction, and analysis of a novel class of self-folding RNA. AB - RNA can play multiple biological roles through use of its three-dimensional (3-D) structures. Recent advances in RNA structural biology have revealed that complex RNA 3D structures are assemblages of double-stranded helices with a variety of tertiary structural motifs. By employing RNA tertiary structural motifs together with the helices, we designed a novel class of self-folding RNA. In RNA composed of three helices (P1, P2, and P3), P1 interacts with P3 via a tetraloop-receptor interaction and P2 forms consecutive base-triples. Two designed RNAs of this class were prepared and their folding properties indicate that they form defined tertiary structures as designed. These RNAs may be used as modular units for constructing artificial ribozymes or nanometer-scale materials. PMID- 11937058 TI - Structural basis for proofreading during replication of the Escherichia coli chromosome. AB - The epsilon subunit of the Escherichia coli replicative DNA polymerase III is the proofreading 3'-5' exonuclease. Structures of its catalytic N-terminal domain (epsilon186) were determined at two pH values (5.8 and 8.5) at resolutions of 1.7 1.8 A, in complex with two Mn(II) ions and a nucleotide product of its reaction, thymidine 5'-monophosphate. The protein structure is built around a core five stranded beta sheet that is a common feature of members of the DnaQ superfamily. The structures were identical, except for differences in the way TMP and water molecules are coordinated to the binuclear metal center in the active site. These data are used to develop a mechanism for epsilon and to produce a plausible model of the complex of epsilon186 with DNA. PMID- 11937059 TI - The structural basis for catalysis and specificity of the Pseudomonas cellulosa alpha-glucuronidase, GlcA67A. AB - Alpha-glucuronidases, components of an ensemble of enzymes central to the recycling of photosynthetic biomass, remove the alpha-1,2 linked 4-O-methyl glucuronic acid from xylans. The structure of the alpha-glucuronidase, GlcA67A, from Pseudomonas cellulosa reveals three domains, the central of which is a (beta/alpha)(8) barrel housing the catalytic apparatus. Complexes of the enzyme with the individual reaction products, either xylobiose or glucuronic acid, and the ternary complex of both glucuronic acid and xylotriose reveal a "blind" pocket which selects for short decorated xylooligosaccharides substituted with the uronic acid at their nonreducing end, consistent with kinetic data. The catalytic center reveals a constellation of carboxylates; Glu292 is poised to provide protonic assistance to leaving group departure with Glu393 and Asp365 both appropriately positioned to provide base-catalyzed assistance for inverting nucleophilic attack by water. PMID- 11937060 TI - Crystal structures of S100A6 in the Ca(2+)-free and Ca(2+)-bound states: the calcium sensor mechanism of S100 proteins revealed at atomic resolution. AB - S100A6 is a member of the S100 family of Ca(2+) binding proteins, which have come to play an important role in the diagnosis of cancer due to their overexpression in various tumor cells. We have determined the crystal structures of human S100A6 in the Ca(2+)-free and Ca(2+)-bound states to resolutions of 1.15 A and 1.44 A, respectively. Ca(2+) binding is responsible for a dramatic change in the global shape and charge distribution of the S100A6 dimer, leading to the exposure of two symmetrically positioned target binding sites. The results are consistent with S100A6, and most likely other S100 proteins, functioning as Ca(2+) sensors in a way analogous to the prototypical sensors calmodulin and troponin C. The structures have important implications for our understanding of target binding and cooperativity of Ca(2+) binding in the S100 family. PMID- 11937061 TI - Rab-subfamily-specific regions of Ypt7p are structurally different from other RabGTPases. AB - The GTPase Ypt7p from S. cerevisiae is involved in late endosome-to-vacuole transport and homotypic vacuole fusion. We present crystal structures of the GDP- and GppNHp-bound conformation of Ypt7p solved at 1.35 and 1.6 A resolution, respectively. Despite the similarity of the overall structure to other Ypt/Rab proteins, Ypt7p displays small but significant differences. The Ypt7p-specific residues Tyr33 and Tyr37 cause a difference in the main chain trace of the RabSF2 region and form a characteristic surface epitope. Ypt7p*GppNHp does not display the helix alpha2, characteristic of the Ras-superfamily, but instead possess an extended loop L4/L5. Due to insertions in loops L3 and L7, the neighboring RabSF1 and RabSF4 regions are different in their conformations to those of other Ypt/Rab proteins. PMID- 11937062 TI - Structure of the lac operon galactoside acetyltransferase. AB - The galactoside acetyltransferase (thiogalactoside transacetylase) of Escherichia coli (GAT, LacA, EC 2.3.1.18) is a gene product of the classical lac operon. GAT may assist cellular detoxification by acetylating nonmetabolizable pyranosides, thereby preventing their reentry into the cell. The structure of GAT has been solved in binary complexes with acetyl-CoA or CoA and in ternary complexes with CoA and the nonphysiological acceptor substrates isopropyl beta-D-thiogalactoside (IPTG) or p-nitrophenyl beta-D-galactopyranoside (PNPbetaGal). A hydrophobic cleft that binds the thioisopropyl and p-nitrophenyl aglycones of IPTG and PNPbetaGal may discriminate against substrates with hydrophilic substituents at this position, such as lactose, or inducers of the lac operon. An extended loop projecting from the left-handed parallel beta helix domain contributes His115, which is in position to facilitate attack of the C6-hydroxyl group of the substrate on the thioester. PMID- 11937063 TI - The crystal structure of diadenosine tetraphosphate hydrolase from Caenorhabditis elegans in free and binary complex forms. AB - The crystal structure of C. elegans Ap(4)A hydrolase has been determined for the free enzyme and a binary complex at 2.0 A and 1.8 A, respectively. Ap(4)A hydrolase has a key role in regulating the intracellular Ap(4)A levels and hence potentially the cellular response to metabolic stress and/or differentiation and apoptosis via the Ap(3)A/Ap(4)A ratio. The structures reveal that the enzyme has the mixed alpha/beta fold of the Nudix family and also show how the enzyme binds and locates its substrate with respect to the catalytic machinery of the Nudix motif. These results suggest how the enzyme can catalyze the hydrolysis of a range of related dinucleoside tetraphosphate, but not triphosphate, compounds through precise orientation of key elements of the substrate. PMID- 11937064 TI - Exercise induces angiogenesis but does not alter movement representations within rat motor cortex. AB - The effects of exercise on the topography of movement representations and blood vessel density within the rat forelimb motor cortex was examined. Adult male rats were allocated to either a Voluntary eXercise (VX) or Inactive Condition (IC). VX animals were housed for 30 days with unlimited access to running wheels while IC animals were housed in standard laboratory cages. VX animals exhibited a progressive increase in the distance traveled per day and ran an average of 58.3 km across the 30-day training period. Microelectrode stimulation was used to derive high resolution maps of the forelimb representations within the motor cortex of animals from both conditions. No significant differences in the area of either distal (wrist/digit) or proximal (elbow/shoulder) movement representations were found between VX and IC animals. However, VX animals did have a significantly greater density of blood vessels within layer V of the forelimb motor cortex. These results demonstrate that increases in forelimb motor activity sufficient to induce cortical angiogenesis does not alter the topography of forelimb movement representations within forelimb motor cortex. PMID- 11937065 TI - Cooling lesions of the lateral parabrachial nucleus during LiCl activation block acquisition of conditioned taste avoidance in male rats. AB - Lesions of the lateral parabrachial nucleus (lPBN) disrupt acquisition of LiCl induced conditioned taste avoidance. Animals with lesions in this area also fail to exhibit taste neophobia. This raises the possibility that an inability of rats to recognize the taste solution as novel contributes to the deficit in taste avoidance learning. If this is the case, then one would expect conditioned taste avoidance not to be disrupted if the lPBN is functional during taste processing but not during LiCl processing. The first three experiments demonstrated that cooling was a viable method by which to temporarily inactivate the lPBN. Measurement of neural temperature during cooling indicated that the lPBN was cooled to temperatures that have been shown to block synaptic transmission but not axonal transmission. Cooling the lPBN itself induced a conditioned avoidance to a sucrose solution but this avoidance was abolished by exposure to daily cooling for 1 week prior to acquisition. In experiment 4, all animals were preexposed to lPBN cooling for 1 week. Those rats that received cooling lesions during a period that started immediately after sucrose solution consumption and extended through the peak effectiveness of LiCl failed to acquire a taste avoidance. These results fail to support the hypothesis that the deficit in taste avoidance learning after permanent lesions of the lPBN is due to an inability of lesioned animals to recognize the taste as novel. They are consistent with the hypothesis that this neural area processes ascending information about LiCl. PMID- 11937066 TI - Effect of hypoxia/hypercapnia on metabolism of 6-[(18)F]fluoro-L-DOPA in newborn piglets. AB - There is evidence that the dopaminergic system is sensitive to altered p(O(2)) in the immature brain. However, the respective enzyme activities have not been measured in the living neonatal brain together with brain oxidative metabolism. Therefore 18F-labelled 6-fluoro-L-3,4-dihydroxyphenylalanine (FDOPA) together with positron emission tomography (PET) was used to estimate the activity of the aromatic amino acid decarboxylase (AADC) in the brain of fifteen newborn piglets (2-5 days old). Two PET scans were performed in each piglet. Eleven animals underwent a period of normoxia and moderate hypoxia/hypercapnia (H/H). The remaining four animals were used as an untreated control group. Simultaneously, the brain tissue p(O(2)) was recorded, the regional cerebral blood flow (CBF) was measured with colored microspheres and the cerebral metabolic rate of oxygen (CMRO(2)) was determined. In addition, in four untreated and six H/H treated piglets the relative amounts of fluorodopamine and the respective metabolites were determined in brain tissue samples using HPLC analysis. H/H conditions were induced by lowering the inspired fraction of oxygen from 0.35 to 0.10 and adding CO(2) to the inspired gas resulting in an arterial p(CO(2)) between 74 and 79 mmHg. H/H elicited a more than 3-fold increase of the CBF (P<0.05) so that the CMRO(2) remained unchanged throughout the H/H period. Despite this, the brain tissue p(O(2)) was reduced from 19+/-4 to 6+/-3 mmHg (P<0.05). The permeability surface area product of FDOPA (PS(FDOPA)) was unchanged. However, the transfer rate of FDOPA (k(3)(FDOPA)) of the nigrostriatal dopaminergic system and the relative amounts of fluorodopamine and the respective metabolites were significantly increased (P<0.05). It is suggested that H/H induces an increase of AADC activity. However, an H/H-induced CBF increase maintains bulk O(2) delivery and preserves CMRO(2). PMID- 11937067 TI - Tonal response patterns of primary auditory cortex neurons in alert cats. AB - The firing rates of primary auditory cortex (A1) neurons are known to be modulated only at the onset, offset, and change of a tonal stimulus in anesthetized animals. The tonal response pattern has been rarely investigated in alert animals. We investigated the time-course of A1 neuron responses to a steady tonal stimulus in alert cats. We found four types of firing responses based on statistical evaluation of the time course of the firing rate. The tonic cells (38 cells) showed a significant (P<0.05) firing increase throughout the stimulus period after a relatively long latency (mean, 25.3 ms) with little tendency of adaptation. The phasic-tonic cells (22 cells) showed a significant firing increase throughout the stimulus period after a medium latency (19.8 ms) with tendency of adaptation to less than a half of the maximum excitation level. Phasic cells (15 cells) responded, after a short latency (10.2 ms), at onset and offset of the stimuli. The unresponsive cells (26 cells) did not show a significant firing increase during stimuli. The findings suggest that there is a functional difference between each type of cells: the tonic cells encode information of static auditory signals in their firing rates; the phasic-tonic cells, of the changing auditory signal during the stimulus period; and the phasic cells, of rapid change of the auditory signal at onset and offset. PMID- 11937068 TI - Phosphorylated neurofilaments and SNAP-25 in cultured SH-SY5Y neuroblastoma cells. AB - Components of the extracellular matrix (ECM) of mammals have profound effects on the behavior and differentiation of many different cell types. Here, we report the results of biochemical and immunocytochemical investigations of the expression of SNAP-25 and phosphorylated neurofilament proteins (NFs) by cells grown on coverslips, cells cultured in EHS-ECM gels, and cells in situ in rat brain. SNAP-25 and phosphorylated NFs were detected by immunofluorescence in all these environments but were not detectable by Western analysis in extracts of cells grown on coverslips. The results support the interpretation that EHS-ECM induces differentiation of SH-SY5Y cells in culture and suggest this system as a model system for study of nerve tissue formation and repair. PMID- 11937069 TI - The distribution of 5-HT(6) receptors in rat brain: an autoradiographic binding study using the radiolabelled 5-HT(6) receptor antagonist [(125)I]SB-258585. AB - We used the highly selective 5-HT(6) receptor radioligand [(125)I]SB-258585 (4 iodo-N-[4-methoxy-3-(4-methylpiperazin-1-yl)phenyl]benzene-sulfonamide) to perform autoradiographic binding studies on the rat brain. High levels of specific binding occurred in the corpus striatum, nucleus accumbens, Islands of Calleja and the olfactory tubercle. A high level of binding also appeared in the choroid plexus. Moderate levels occurred in several regions of the hippocampal formation and in certain regions of the cerebral cortex, thalamus, hypothalamus, and substantia nigra; and very low levels in the globus pallidus, cerebellum, other mesencephalic regions, and the rhombencephalon. Displacement of total binding with 10 microM unlabelled SB-214111 (4-bromo-N-[4-methoxy-3-(4 methylpiperazin-1-yl)phenyl]benzene-sulfonamide), another selective 5-HT(6) receptor antagonist, or 10 microM unlabelled methiothepin, reduced binding to barely discernible levels. Some animals received unilateral injections of 6 hydroxydopamine (6-OHDA) into the median forebrain bundle to lesion the nigro striatal pathway before autoradiographic examination. Effectiveness of the 6-OHDA lesions in the substantia nigra and striatum was confirmed with tyrosine hydroxylase immunohistochemistry. Such lesions resulted in no significant changes in [(125)I]SB-SB258585 binding in any brain region examined, suggesting that 5 HT(6) receptors in the striatum are not located on dendritic, somatic or terminal elements of dopaminergic neurones. Thus, the striatal binding sites seen in this study may be on intrinsic GABAergic or cholinergic neurones, or on terminals of projection neurones from the thalamus or cerebral cortex. The 5-HT(6) receptor ligand binding seen here in the striatum, accumbens, olfactory tubercle, Islands of Calleja, cerebral cortex and hippocampus are in concordance with previous immunohistochemical studies, and suggest a possible involvement of 5-HT(6) receptors in locomotor control, cognition, memory, and control of affect. The high levels of binding observed in the choroid plexus in this study have not been reported before. This finding suggests that 5-HT(6) receptors could play a role in the control of cerebrospinal fluid dynamics. PMID- 11937070 TI - Spontaneous activity in rat vestibular nuclei in brain slices and effects of acetylcholine agonists and antagonists. AB - Extracellular recording was used to investigate spontaneously active neurons in all four major nuclei of the rat vestibular nuclear complex (VNC) in brainstem slices. The density of spontaneously active neurons was highest in the medial vestibular nucleus (MVN), slightly lower in the superior (SuVN) and spinal (SpVN) nuclei, and lowest in the lateral vestibular nucleus (LVN). We compared the effects of acetylcholine agonists and antagonists on spontaneously discharging neurons in MVN, SuVN, and SpVN with those in the nearby dorsal cochlear nucleus (DCN). The proportion of neurons responding to carbachol was greatest in DCN and smallest in SpVN. Unlike in DCN, some neurons in MVN, SuVN, and SpVN showed decreased firing during carbachol or muscarine. Magnitudes of responses to carbachol and muscarine were closely correlated (P<0.01). MVN neurons possessed nicotinic as well as muscarinic receptors. Activation of either type was unaffected by blocking synaptic transmission. The IC(50) values for the muscarinic subtype-preferential antagonists were compared, and tropicamide, preferential for M(4), was the most potent. Our results suggest that: (1) the relative numbers of spontaneously active neurons in rat VNC differ among nuclei; (2) acetylcholine agonists elicit changes in mean firing rates of neurons in MVN, SuVN and SpVN, but fewer neurons respond, and responses are smaller than in DCN; (3) both muscarinic and nicotinic acetylcholine receptors are present on MVN neurons, but muscarinic receptors may be more prominent. PMID- 11937071 TI - A role for protein kinase C and its substrates in the action of valproic acid in the brain: implications for neural plasticity. AB - Valproic acid (VPA) is a broad-spectrum anticonvulsant with well-documented teratogenic effects, but whose mechanism of action is largely unknown. In the present study we have examined the effects of VPA on the expression of two prominent substrates for protein kinase C (PKC) in the brain, MARCKS and GAP-43, which have been implicated in actin-membrane plasticity and neurite outgrowth during neuronal differentiation, respectively, and are essential to normal brain development. Immortalized hippocampal HN33 cells exposed to VPA exhibited reduced MARCKS protein expression and demonstrated increased GAP-43 protein expression, with concomitant alterations in cellular morphology, including an increase in the number and length of neurites and accompanied by a reduction in cell growth rate. The effects of VPA were observed at clinically relevant concentrations following chronic (>1 day) VPA exposure. We also present evidence for a VPA-induced alteration in PKC activity, as well as temporal changes in individual PKC isozyme expression. Inhibition of PKC with the PKC-selective inhibitor, LY333531, prevented the VPA-induced down-regulation of membrane-associated MARCKS, but had no effect on the cytosolic MARCKS reduction or the GAP-43 up-regulation. Inhibition of PKC by LY333531 enhanced the differentiating effects of VPA; additionally, LY333531 alone induced greater neurite outgrowth in this cell line. Collectively, these data indicate that VPA induces neuronal differentiation, associated with a reduction in MARCKS expression and an increase in GAP-43 expression, consistent with the hypothesis that a reduction in MARCKS at the membrane may be permissive for cytoskeletal plasticity during neurite outgrowth. PMID- 11937072 TI - Effect of ionotropic glutamate receptor antagonists on Fos-like immunoreactivity in the dorsal horn following transection of the rat sciatic nerve. AB - Fos-like immunoreactivity (FLI) was investigated in the lumbar dorsal horn 2 h after transection of the rat sciatic nerve and sham operation. FLI following nerve transection was distributed through the medio-lateral extension of the superficial layer of the dorsal horn, while FLI after sham operation, tissue injury, was restricted to the lateral one-third of this layer. The number of FLI neurons in the lateral one-third was similar in the two operations, indicating that neurons expressing FLI in the medial two-thirds and in the lateral one-third of the superficial layer after nerve transection are derived from nerve injury and tissue injury, respectively. FLI in the lateral one-third, but not the medial two-thirds, after nerve transection was significantly reduced by pretreatment with NMDA and AMPA/KA receptor antagonists, indicating that there is a considerable difference in the contributions of ionotropic glutamate receptors to FLI in this layer induced by nerve injury and tissue injury. PMID- 11937074 TI - Cardiac markers of acute coronary syndromes: is there a case for point-of-care testing? AB - OBJECTIVE: Major challenges for physicians include selection of effective tests in the time-sensitive identification and management of patients with acute coronary syndromes (ACS). We review whether cardiac marker testing performed at the point-of-care (POC) has an impact on clinical management and guidance of intervention for ACS patients. DESIGN AND METHODS: Evidence from recently published studies and meta-analyses supports the efficacy of cardiac markers. Technologies and specifications of all currently available POC tests for monitoring cardiac markers are surveyed. Finally, a series of questions to investigate the utility of cardiac markers, and their measurement by POC tests, for clinical management and guidance of therapy for ACS patients, are addressed. RESULTS: Cardiac troponins are clearly the best markers for the definitive detection of myocardial infarction. Compelling evidence for the utility of troponins in risk stratification and guidance of intervention for ACS patients has resulted in inclusion of cardiac markers in clinical guidelines. Rapid multi analyte POC tests, few of which exhibit harmony with central laboratory assays, have facilitated the use of cardiac markers for clinical management and guidance of therapy. CONCLUSIONS: Given the need to minimize vein-to-brain time, it is expected that point-of-care testing of cardiac markers will take a leading role in management of ACS patients. PMID- 11937073 TI - Caveats in hair analysis in chronic arsenic poisoning. AB - The diagnosis and evaluation of chronic arsenic poisoning remains a difficult task. Clinical indicators are crude measures, and electromyography adds little to the picture. Blood arsenic levels are transitory, however urine levels are useful for monitoring ongoing exposure. Hair arsenic is useful as a confirmatory feature in chronic arsenic poisoning provided external contamination by arsenic can be excluded. The distribution of arsenic in cross sections or along the length of a shaft of hair cannot distinguish external contamination from arsenic derived from ingestion. PMID- 11937075 TI - Comparison of near infrared reflectance analysis of fecal fat, nitrogen and water with conventional methods, and fecal energy content. AB - OBJECTIVES: To evaluate Near-Infrared Analysis (NIRA) method for determining fecal fat, water and nitrogen. DESIGN AND METHODS: The results of fecal fat, water and nitrogen by NIRA were compared with results of van de Kamer and Acid Steatocrit (AS), Dumas and vacuum drying methods for fat, nitrogen and water respectively. Results of fat determining methods were also compared with total fecal energy as obtained by bomb calorimeter. RESULTS: NIRA results correlated significantly (p < 0.001) with standard methods for nitrogen (r = 0.79), fat (r = 0.84 and r = 0.88 for van de Kamer and AS respectively) and water (r = 0.91). The limits of agreement for nitrogen and fat results were too wide for the methods to be used interchangeably. The fecal fat results correlated significantly (p < 0.001) with fecal energy results. CONCLUSION: NIRA may be valuable for monitoring malabsorption but the diagnostic value remains to be determined. PMID- 11937076 TI - A simultaneous LightCycler detection assay for five genetic polymorphisms influencing drug sensitivity. AB - OBJECTIVES: The routine detection of polymorphisms affecting drug sensitivity in patients before treatment is important in the identification of drug responders or nonresponders, and patients at increased risk of drug toxicity. Here, we present an assay for the simultaneous and rapid genotyping of five polymorphisms influencing drug sensitivity. DESIGN AND METHODS: We used a hybridization probe assay on the LightCycler to detect five single nucleotide polymorphisms (SNPs): INPP1 (973C>A), ADRB2 (R16G and Q27E), HTR2A (102T>C), and mtDNA (1555A>G). Two fluorescent labeled hybridization probes were designed for the simultaneous detection of the five SNPs and detection of the variant alleles was performed by melting curve analysis. RESULTS: All five SNPs were detected with a single thermocycle protocol within 40 min. The genotypes determined in this assay were identical to those obtained with conventional PCR and restriction fragment length polymorphism analysis. CONCLUSIONS: To our knowledge, we report here for the first time a method for simultaneous detection of five SNPs, on a single thermocycle protocol by the LightCycler. This method is rapid, highly sensitive, and high-throughput, and is thus suitable for routine clinical use and large scale epidemiologic studies. PMID- 11937077 TI - Semi-automated enzymatic measurement of serum zinc concentration. AB - OBJECTIVE: To measure serum zinc concentration by means of carbonic anhydrase reactivation using an automated analyzer. METHODS: The zinc content of carbonic anhydrase (CA), whose cofactor is zinc, was removed by dialysis against pyridine 2 to 6 dicarboxylic acid and a pure apoenzyme was obtained. Serum proteins were precipitated with trichloroacetic acid (TCA) solution and the supernatant fraction of the sample was used to determine the zinc concentration. The negative effects of the precipitant on CA activity in the assay were completely removed, reaction conditions for maximal CA activity were provided and the color of the product was enhanced and stabilized. P-nitrophenyl acetate was used as the substrate and the change of absorbance of p-nitrophenol which was produced was followed at 400 nm. The initial rate of the esterase activity of CA was measured by using an automated analyzer. Analytical performance characteristics of the assay were determined. The zinc concentrations in serum samples of healthy subjects and patients were measured. RESULTS: The enzymatic assay is accurate, sensitive, specific and is not affected by other metals. There was excellent agreement with the results obtained using an atomic absorption spectrophotometer (AAS) (y = 0.98X + 0.18, r = 0.99). Serum zinc concentrations were found to be higher in patients with vivax malaria, and lower in patients with cutaneous leishmaniasis than in healthy subjects. CONCLUSION: The enzymatic method is suitable for semiautomated measurement of serum zinc concentration. PMID- 11937078 TI - Validity of PCR with emphasis on variable number of tandem repeat analysis. AB - OBJECTIVES: Variable number of tandem repeat polymorphisms (VNTR) are frequently analyzed by PCR in genetic, epidemiologic and forensic studies. We wanted to explore the validity of these PCR analyses. DESIGN AND METHODS: The amplification of the different alleles of the 17- and the 44-bp VNTR of the serotonin transporter gene and the 39-bp VNTR of the glycoprotein Ibalpha gene was analyzed. We studied the effects of the parameters magnesium, dimethylsulfoxide, 7-deaza-dGTP, formamide, betaine, PCR temperatures and different types of polymerases. RESULTS: In all three VNTR polymorphisms selective amplification of one of the alleles of heterozygous individuals could be obtained by change of the magnesium concentration. This problem could be minimized by a combination of Taq- and Pwo-polymerases and by use of 7-deaza-dGTP. CONCLUSION: PCR analysis of all of these VNTRs may give reproducibly wrong results in truly heterozygous subjects due to selective amplification of only one of the alleles. PMID- 11937079 TI - Mechanism of Impila (Callilepis laureola)-induced cytotoxicity in Hep G2 cells. AB - OBJECTIVES: To determine the mechanism(s) of Impila (Callilepis laureola)-induced toxicity in human hepatoblastoma Hep G2 cells in vitro and the possible prevention of this toxicity by N-acetylcysteine (NAC). DESIGN AND METHODS: Cells were treated with an aqueous extract of Impila (10 mg/mL) for up to 24 h. NAC (5 mM) was administered either concomitantly with Impila or one hour post Impila treatment. Cytotoxicity was quantitated spectrophotometrically by the metabolism of the tetrazolium dye MTT. Total glutathione (GSH) was measured using the Tietze assay. RESULTS: Impila produced cytotoxicity and depleted GSH in a concentration- and time-dependent manner. A significant depletion in GSH was observed after 15 min (p < 0.0001 vs. control), whereas significant cytotoxicity was only observed after at least 3 h (p < 0.0001 vs. control). Both concomitant and posttreatment with NAC prevented Impila-induced GSH depletion and resulted in a significant decrease in Impila-induced cytotoxicity (p < 0.001 vs. NAC-untreated cells). CONCLUSION: Our results suggest the mechanism of Impila-induced cytotoxicity in Hep G2 cells in vitro involves depletion of cellular GSH. Preventing GSH depletion by supplementing cells with NAC reduces cytotoxicity. PMID- 11937080 TI - Tamm-Horsfall protein excretion to predict the onset of renal insufficiency. AB - OBJECTIVE: Immunosuppressive therapy after liver transplantation may be a risk for kidney dysfunction. This work was designed to determine whether Tamm-Horsfall Protein (THP) could be considered as a marker for nephrotoxicity. DESIGN AND METHODS: THP was determined by an ELISA method in serial 24-h urine from liver transplant patients. Fourteen patients suffered renal insufficiency (LTr(1)) and 20 showed no acute renal damage (LTr(2)) after liver transplantation. RESULTS: No clear association could be seen between daily THP excretion and plasma creatinine levels by comparing serial samples collected at the same time. Nevertheless, significant differences were observed in pretransplant THP excretion between both groups of patients. The results (Median/Interquartile Range) were: CONTROLS: 113.2/84.9 to 146.8 mg/24 h (n = 30); LTr(1): 36.9/18.3 to 54.5 mg/24 h (p<<0.001 with respect to C and LTr(2)); LTr(2): 90.8/61.5 to 139.7 mg/24 h. CONCLUSIONS: The higher pretransplant synthesis and/or secretion of THP seem to have a protective role on the kidney during and after liver transplantation. PMID- 11937081 TI - Lens superoxide dismutase and catalase activities in diabetic cataract. AB - OBJECTIVE: Biochemical evidence suggests that the oxidative damage of the lens proteins is involved in the genesis of senile cataract and the development of diabetes-related pathologic changes such as the formation of cataracts. In particular, lens proteins are subject to extensive oxidative modification. Oxidative damage either decreases the antioxidant capacity or decreased antioxidant capacity results in oxidative damage. The purpose of this study was to analyze the activities of the antioxidant enzymes such as Cu,Zn Superoxide Dismutase (Cu,Zn-SOD) and catalase in the cataractous lenses of the type 2 diabetic group and cataractous lenses of the senile group. METHOD: Eighteen diabetic cataractous lenses and twenty six senile cataractous lenses were studied. Cu,Zn-SOD activity was measured in lenses by enzymatic method and catalase activity was measured by colorimetric method. RESULTS: Cu,Zn-SOD levels were significantly lower in the diabetic cataractous lenses than senile cataractous lenses (respectively 8.052 +/- 0.818, 18.216 +/- 4.217 microg/g prot. p < 0.05). Similarly, catalase levels were significantly lower in the diabetic cataractous lenses than senile cataractous lenses (respectively 0.326 +/- 0.134, 0.665 +/- 0.322 kU/g prot. p < 0.001). CONCLUSION: The results of the present study indicate that the antioxidant capacity in the diabetic cataractous lenses were decreased and this result suggests a role of antioxidant enzymes in the genesis of diabetic cataracts. PMID- 11937082 TI - Decreased anion gap in polyclonal hypergammaglobulinemia. AB - OBJECTIVES: The anion gap has proved a valuable tool in the diagnosis of various forms of acid-base disorders, although the importance of slight rises in the anion gap remains unclear. The concept of the anion gap is often misunderstood and misapplied. The relationship between gammaglobulins and the serum anion gap has not received much attention except for reports of a narrowing of the gap associated with certain monoclonal immunoglobulin G gammopathies. We present patients with polyclonal gammopathy, the magnitude of which correlated strongly and negatively with the anion gap. DESIGN: The anion gap can be readily calculated from routine laboratory data, and anion gap was calculated as ([Na] +[K])- ([Cl] + [HCO3]). SUBJECTS: Serum anion gaps were determined in 206 patients with polyclonal hypergammaglobulinemia and 63 healthy subjects. MEASUREMENTS: Serum sodium and potassium ions concentration were determined by flame photometry. Serum bicarbonate level was measured as total carbon dioxide content. Serum chloride level was determined by chlorimetric titration with silver ions. RESULTS: All patients with polyclonal hypergammaglobulinemia had a statistically significant reduction in their mean serum anion gaps (6.4 +/- 1.2 mmol/L) when compared with normal control volunteers (15.3 +/- 2.4 mmol/L), p < 0.05. CONCLUSIONS: Our results show a negative correlation between serum anion gap and gammaglobulins concentration. PMID- 11937083 TI - Circulating monocyte chemoattractant protein-1 in heart transplant recipients with cardiac allograft vasculopathy. AB - OBJECTIVE: To evaluate serum Monocyte Chemoattractant Protein-1 (MCP-1) as a biologic marker of cardiac allograft vasculopathy (CAV) in heart transplant recipients (HTR). DESIGN AND METHODS: Serum levels of MCP-1 were measured in 49 HTR with and without CAV. RESULTS: HTR exhibited significantly higher serum MCP-1 levels than controls. However, no difference was observed according to the existence of CAV. CONCLUSION: Serum MCP-1 levels do not constitute a marker of the CAV occurring after heart transplantation. PMID- 11937084 TI - Ubiquinone-10 content in lymphocytes of phenylketonuric patients. AB - OBJECTIVES: To investigate the ubiquinone-10 content in lymphocytes from phenylketonuric patients. DESIGN AND METHODS: We compared 23 patients with 25 age matched controls. Ubiquinone-10 was analyzed by HPLC with electrochemical detection. RESULTS: Ubiquinone-10 concentrations were significantly lower in patients (77-270 nmol/g of protein) compared with controls (190-550) (p < 0.001). Significantly negative correlation was observed between ubiquinone-10 and phenylalanine (r = -0.441; p < 0.05). CONCLUSIONS: Ubiquinone-10 concentrations are decreased in lymphocytes from phenylketonuric patients. This deficiency is associated with high plasma phenylalanine concentrations. PMID- 11937086 TI - Is the 5-HT(7) receptor involved in the pathogenesis and prophylactic treatment of migraine? AB - The mechanisms underlying the pathogenesis of migraine and their possible association with serotonin (5-hydroxytryptamine; 5-HT) have not yet been elucidated. One of the major obstacles in achieving this goal is the lack of information on the mechanisms by which the monoamine could possibly trigger and/or modulate the basic pathophysiological features of the condition, that is, cranial vasodilatation and neurogenic inflammation. This information should provide a useful theoretical framework to insight the nature of the postulated fundamental triggering mechanism in the brain that ultimately results in head pain. Novel avenues for research and drug development may be envisaged upon the recent observations showing that 5-HT is actually able to produce vasodilatation of intra- and extra-cranial blood vessels through a mechanism pharmacologically resembling the 5-HT(7) receptor type, and that the messenger RNA (mRNA) encoding for this receptor is highly expressed in cranial vessels. Other lines of evidence have suggested that the 5-HT(7) receptor may play an excitatory role in neuronal systems and that it may be involved in hyperalgesic pain and neurogenic inflammation. On the basis of these observations, it is proposed that the 5-HT(7) receptor may well represent a link between the abnormal phenomena of 5-HT processing and neurotransmission that are observed in migraine patients, and the vascular and neurogenic alterations that account for migraine headache. This view is supported by the fact that most of the migraine prophylactic 5-HT receptor antagonists display relatively high affinity for the 5-HT(7) receptor, which significantly correlates with their pharmaceutically active oral doses. PMID- 11937087 TI - Evidence for changes in the tachyphylactic property of recombinant angiotensin II AT(1) receptor expressed in CHO cells. AB - The manifestation of tachyphylaxis to angiotensin II in Chinese hamster ovary (CHO) cells expressing the rat angiotensin II AT(1) receptor was investigated. The cells were transfected with a cDNA fragment containing the complete coding region of the angiotensin II AT(1A) receptor gene, as well as 56 bp of its 3'- and 52 bp of its 5'-untranslated regions. These cells (CHO-AT(1)) responded to angiotensin II by increases in intracellular Ca(2+) concentration and inositol phosphate turnover, which were inhibited upon repeated administrations, characterizing the tachyphylaxis phenomenon. In contrast to smooth muscle cells, which are rendered tachyphylactic to angiotensin II but not to [2 lysine]angiotensin II ([Lys(2)]angiotensin II), this analogue induced responses in CHO-AT(1) cells that were also inhibited upon repeated administrations. A smooth muscle cell line, which showed tachyphylaxis only to angiotensin II, became tachyphylactic also to [Lys(2)]angiotensin II after transfection with the angiotensin II AT(1) receptor gene. Our findings suggest that posttranscriptional control directed by the 3'- or the 5'-untranslated regions in the angiotensin II AT(1) receptor gene may play a role in modulating the signal transduction pathways involved in the mechanism of angiotensin II tachyphylaxis. PMID- 11937088 TI - Anisodamine inhibits cardiac contraction and intracellular Ca(2+) transients in isolated adult rat ventricular myocytes. AB - Increased cardiac workload often leads to serious complications during cardiac surgery such as pericardiopulmonary bypass. Various agents have been applied to lower peripheral resistance and cardiac workload, one of which, anisodamine, is widely used in Asia. However, the direct action of anisodamine on cardiac contractile property is essentially unknown. This study was designed to examine the influence of anisodamine on ventricular contractile function at the single cardiac myocyte level. Ventricular myocytes from adult rat hearts were stimulated to contract at 0.5 Hz, and mechanical and intracellular Ca(2+) properties were evaluated using an IonOptix Myocam system. Contractile properties analyzed included peak shortening (PS), time-to-PS (TPS), time-to-90% relengthening (TR(90)), maximal velocity of shortening/relengthening (+/-dL/dt), intracellular Ca(2+) fluorescence intensity change (DeltaFFI) and decay (tau). Anisodamine exhibited a concentration-dependent (10(-12)-10(-6) M) inhibition in PS and DeltaFFI, with maximal inhibitions of 44.7% and 47.2%, respectively. Anisodamine inhibited +/-dL/dt, lowered resting FFI but elicited no effect on TPS/TR(90) and tau. Pretreatment with the nitric oxide synthase (NOS) inhibitor N(omega)-nitro-L arginine methyl ester (L-NAME, 100 microM) abolished the inhibitory effect of anisodamine in cell shortening. In addition, anisodamine prevented cholinoceptor agonist carbachol-induced positive cardiac contractile response. This study demonstrated a direct cardiac depressive action of anisodamine at the myocyte level, which may be related to, at least in part, NO production and cholinoceptor antagonism. PMID- 11937089 TI - Activation of nitric oxide synthase through muscarinic receptors in rat parotid gland. AB - Muscarinic receptors play an important role in secretory and vasodilator responses in rat salivary glands. Nitric oxide synthase (NOS) activity was found coupled to muscarinic receptor activation as well as to nitric oxide-mediated amylase secretion elicited by carbachol. Parotid glands presented a predominant M(3) and a minor muscarinic M(1) acetylcholine receptor population, though carbachol stimulated NOS activity only through muscarinic M(3) receptors as revealed in the presence of 4-diphenylacetoxy-N-methylpiperidine methiodide (4 DAMP) and pirenzepine. Amylase secretion induced by carbachol appeared to be partly mediated by nitric oxide and nitric oxide-induced signaling since N-nitro L-arginine methyl ester (L-NAME) inhibited the effect as well as did methylene blue. A negative regulation of NOS by protein kinase C activation in the presence of a high concentration of carbachol was seen in parotid glands and this inhibition was paralleled by amylase secretion. PMID- 11937090 TI - The effect of a Titanocene Dichloride derivative, Ti IV (C5H5)(2) NCS(2), on the haematopoietic response of Ehrlich tumour-bearing mice. AB - The effects of the [Ti IV (C(5)H(5))(2) NCS(2)] metallocene (BCDT), a Titanocene Dichloride derivative, on the growth and differentiation of granulocyte macrophage progenitor cells [colony-forming unit-granulocyte-macrophage (CFU-GM)] and bone marrow cellularity in normal and Ehrlich ascites tumour-bearing mice were studied. As expected for the Ehrlich ascites tumour-model, concomitant myelosuppression, increased number of spleen CFU-GM and changes in bone marrow cellularity were observed. The treatment of Ehrlich ascites tumour-bearing mice with BCDT (10-30 mg/kg/day) produced a dose-dependent increase in myelopoiesis, a reduction in splenic colonies and a restoration in the total and differential marrow cell counts. We also observed an increase in CFU-GM number when bone marrow cells obtained from normal mice were incubated in vitro with serum from normal mice treated with BCDT. In addition, BCDT prolonged, in a dose-dependent manner, the survival of mice inoculated with Ehrlich ascites tumour. Although it has been previously reported that substitutions in the two halides of the titanocene do not interfere with antitumoural effect, our results with BCDT demonstrated a reduction in antitumour efficacy when compared to previous results with the original titanocene produced in our laboratory. PMID- 11937091 TI - Pharmacological properties of YM-57029, a novel platelet glycoprotein IIb/IIIa antagonist. AB - The pharmacological properties of YM-57029 [4-[4-(4-carbamimidoylphenyl)-3 oxopiperazin-1-yl]piperidino]acetic acid monohydrochloride trihydrate), a novel glycoprotein IIb/IIIa antagonist were examined in this study. YM-57029 inhibited fibrinogen binding to purified glycoprotein IIb/IIIa, 1000-fold more potently than the tetrapeptide arginine-glycine-aspartic acid-serine (RGDS). YM-57029 concentration-dependently inhibited ADP-, collagen- and high shear stress-induced platelet aggregation, strongly inhibited ATP release from platelets activated by ADP, and enhanced deaggregation of ADP-induced platelet aggregates. In a pro aggregatory activity study, RGDS or SC-54701A ((S)-3-[3-[(4 amidinophenyl)carbamoyl]propionamido]-4-pentynoic acid monohydrochloride) caused prominent small aggregate formation. At a higher concentration, RGDS induced medium and large size aggregates, and SC-54701A induced medium aggregates. In contrast, YM-57029 produced only a few small and no larger size aggregates. Ex vivo ADP-induced platelet aggregation and platelet retention to collagen-coated plastic beads were dose-dependently inhibited by YM-57029 after intravenous bolus injection in guinea pigs. YM-57029 produced dose-dependent antithrombotic effects in carotid artery thrombosis and arterio-venous shunt thrombosis models in guinea pigs at 10 and 30 microg/kg, respectively. At these doses, YM-57029 prolonged template bleeding time. These results suggest that YM-57029 is a potent glycoprotein IIb/IIIa antagonist which has less pro-aggregatory effect. It may be a promising antiplatelet agent for thromboembolic diseases, and a good prototype for developing an orally active compound. PMID- 11937092 TI - Ethanol-mediated inhibition of mitogen-activated protein kinase phosphorylation in mouse brain. AB - Ethanol (1.5-3.5 g/kg body weight) was administered intraperitoneally to mice and the phosphorylation of MAP (mitogen-activated protein) kinase in the cerebral cortex was determined using phospho-specific MAP kinase antibodies. Ethanol inhibited the phosphorylation of MAP kinase in a dose- and time-dependent manner. Developmental studies demonstrated that the levels of phospho-MAP kinase increased from fetal cortex (prenatal) to 16-day-old mice (postnatal) and remained constant up to 4 months of age. However, ethanol (3.5 g/kg) decreased the phospho-MAP kinase staining in all of the age groups studied. Subcellular fractionation studies demonstrated that ethanol inhibited the phosphorylation of MAP kinase in both the cytosol as well as nucleus, but did not alter the levels of MAP kinase. Likewise, MK-801 (0.4 mg/kg) or flurazepam (75 mg/kg) also decreased the phospho-MAP kinase content. These data indicate that ethanol may inhibit the phosphorylation of MAP kinase in vivo by either inhibiting NMDA receptors or activating GABA receptors. PMID- 11937093 TI - Lack of persistent changes in the dopaminergic system of rats withdrawn from methamphetamine self-administration. AB - A continuing challenge for studies in the neurobiology of drug abuse is to identify and characterize long-lived neuroadaptations that can trigger craving and relapse. We previously reported that rats that had actively self-administered methamphetamine for 5 weeks and were then withdrawn from methamphetamine for 24 h showed marked decreases in somatodendritic dopamine D(2) autoreceptor levels in the ventral tegmental area and median and dorsal part of the substantia nigra zona compacta with a corresponding down-regulation of dopamine D(1) receptors in the shell of the nucleus accumbens. The purpose of the present study was to determine whether neuroadaptive changes in dopamine receptors or transporters in the brains of rats withdrawn for 24 h from chronic methamphetamine self administration are persistent changes that can be demonstrated long after withdrawal. A "yoked" procedure was used in which rats were tested simultaneously in groups of three, with only one rat actively self-administering methamphetamine while the other two received yoked injections of either methamphetamine or saline. In vitro quantitative autoradiography was used to determine densities of dopamine uptake sites and dopamine D(1) and D(2) receptors in different brain regions following 7- and 30-day periods of withdrawal from chronic methamphetamine self-administration. No changes in dopamine transporter and dopamine receptor numbers were detected in any brain region examined in rats self administering methamphetamine compared with littermates receiving yoked infusions of either methamphetamine or saline. Thus, neuroadaptive changes in densities of dopamine receptors or transporters in certain brain areas may contribute to the reinforcing effects of methamphetamine during the acquisition and maintenance phases of self-administration, but do not appear to contribute to the long lasting neuroadaptive effects of chronic methamphetamine self-administration, which may trigger craving and relapse. PMID- 11937094 TI - Interaction between vanilloid and glutamate receptors in the central modulation of nociception. AB - This study investigates the effect of microinjections of capsaicin in the periaqueductal grey matter of rats on nociceptive behaviour and the possible interactions with NMDA and mGlu receptors. Intra-periaqueductal grey microinjection of capsaicin (1-3-6 nmol/rat) increased the latency of the nociceptive reaction in the plantar test. This effect was prevented by pretreatment with capsazepine (6 nmol/rat), which had no effect per se on the latency of the nociceptive reaction. 7-(Hydroxyimino)cyclopropa[b]chromen-1alpha carboxylate ethyl ester (CPCCOEt, 50 nmol/rat) and 2-Methyl-6 (phenylethynyl)pyridine (MPEP, 50 nmol/rat), antagonists of mGlu(1) and mGlu(5) receptors, respectively, completely blocked the effect of capsaicin. Similarly, pretreatment with DL-2-Amino-5-phosphonovaleric acid (DL-AP5, 5 nmol/rat) and riluzole (4 nmol/rat), an NMDA receptor antagonist and a voltage-dependent Na(+) channels blocker which inhibits glutamate release, respectively, completely antagonized the effect of capsaicin. However, pretreatment with (2S)-alpha Ethylglutamic acid (30 nmol/rat) and (RS)-alpha-Methylserine-O-phosphate (MSOP, 30 nmol/rat), antagonists of group II and group III mGlu receptors, respectively, had no effects on capsaicin-induced analgesia. Similarly, pretreatment with N (piperidin-1-yl)-5-(4-chlophenyl)-1-(2,4-dichlorophenyl)-4-methyl-1H-pyrazole-3 carboxamide (SR 141716A, 5 pmol/rat), a selective cannabinoid CB(1) receptor antagonist, did not affect the capsaicin-induced antinociception. In conclusion, this study shows that capsaicin might produce antinociception at the periaqueductal grey level by increasing glutamate release, which activates postsynaptic group I mGlu and NMDA receptors. PMID- 11937095 TI - Comparison of nerve conduction blocks by an opioid and a local anesthetic. AB - The experiments were done on frog sciatic nerves, using a sucrose-gap recording technique. The aim of our study was to investigate and to compare the tonic and phasic conduction blocking potency of tramadol and lidocaine on whole nerve and their interactions with Ca(2+). The concentration of a tramadol solution producing the same amount of tonic and phasic conduction blocks was three and six times higher than that needed for lidocaine, respectively. Increasing the Ca(2+) concentration in the test solution enhanced the conduction blocking potency of tramadol, but decreased that of lidocaine. It is concluded that tramadol blocks nerve conduction like a local anesthetic but with a weaker effect than that of lidocaine. Interactions of Ca(2+) and these drugs suggested that these drugs might have either different binding sites or different action mechanisms. PMID- 11937096 TI - Arvanil-induced inhibition of spasticity and persistent pain: evidence for therapeutic sites of action different from the vanilloid VR1 receptor and cannabinoid CB(1)/CB(2) receptors. AB - Activation of cannabinoid receptors causes inhibition of spasticity, in a mouse model of multiple sclerosis, and of persistent pain, in the rat formalin test. The endocannabinoid anandamide inhibits spasticity and persistent pain. It not only binds to cannabinoid receptors but is also a full agonist at vanilloid receptors of type 1 (VR1). We found here that vanilloid VR1 receptor agonists (capsaicin and N-N'-(3-methoxy-4-aminoethoxy-benzyl)-(4-tert-butyl-benzyl)-urea [SDZ-249-665]) exhibit a small, albeit significant, inhibition of spasticity that can be attenuated by the vanilloid VR1 receptor antagonist, capsazepine. Arvanil, a structural "hybrid" between capsaicin and anandamide, was a potent inhibitor of spasticity at doses (e.g. 0.01 mg/kg i.v.) where capsaicin and cannabinoid CB(1) receptor agonists were ineffective. The anti-spastic effect of arvanil was unchanged in cannabinoid CB(1) receptor gene-deficient mice or in wildtype mice in the presence of both cannabinoid and vanilloid receptor antagonists. Likewise, arvanil (0.1-0.25 mg/kg) exhibited a potent analgesic effect in the formalin test, which was not reversed by cannabinoid and vanilloid receptor antagonists. These findings suggest that activation by arvanil of sites of action different from cannabinoid CB(1)/CB(2) receptors and vanilloid VR1 receptors leads to anti spastic/analgesic effects that might be exploited therapeutically. PMID- 11937097 TI - The effects of selective serotonin reuptake inhibitors on extracellular 5-HT levels in the hippocampus of 5-HT(1B) receptor knockout mice. AB - The effects of two selective serotonin reuptake inhibitors on 5-hydroxy tryptamine (5-HT) in the hippocampus were studied in wildtype and in 5-HT(1B) receptor knockout mice using in vivo microdialysis. Basal 5-HT levels in the hippocampus were not different between the two genotypes. The functional absence of 5-HT(1B) receptors was examined in the knockout mice by local infusion of the 5-HT(1B) receptor agonist, 1,4-Dihydro-3-(1,2,3,6-tetrahydro-4-pyridinyl)-5H pyrrolo[3,2-b]pyridin-5-one (CP93129) into the hippocampus. CP93129 (1 microM) decreased 5-HT levels in wildtype mice, but not in 5-HT(1B) knockout mice. Systemic administration of the selective 5-HT reuptake inhibitor paroxetine (5 mg/kg, i.p.) increased extracellular 5-HT levels. The increase of 5-HT in 5 HT(1B) knockout mice was almost twofold higher than in wildtype mice. Systemic administration of selective 5-HT reuptake inhibitors stimulates both terminal 5 HT(1B) autoreceptors and somatodendritic 5-HT(1A) autoreceptors. Therefore, the selective 5-HT reuptake inhibitor, fluvoxamine, was applied locally into the hippocampus to investigate the role of the terminal 5-HT(1B) autoreceptors. Local administration of 0.3 microM fluvoxamine resulted in comparable increases in extracellular 5-HT in both genotypes, whereas 1.0 microM fluvoxamine produced a twofold greater increase in 5-HT levels in 5-HT(1B) knockout as compared to wildtype mice. In conclusion, the differences in hippocampal 5-HT output between wildtype and 5-HT(1B) knockout mice after local or systemic administration of selective 5-HT reuptake inhibitors show that 5-HT(1B) autoreceptors play a significant role in the inhibition of 5-HT release at serotonergic nerve terminals. In addition, the different dose-response to fluvoxamine suggests that 5-HT(1B) knockout mice have possible adaptations of 5-HT transporters in order to compensate for the loss of the terminal 5-HT(1B) autoreceptor. PMID- 11937098 TI - The new antiepileptic drug levetiracetam normalises chlordiazepoxide withdrawal induced anxiety in mice. AB - Some antiepileptic drugs have been used with success to counteract withdrawal symptoms following chronic use of sedatives, hypnotics or alcohol. We evaluated the potential of levetiracetam (Keppra), a new antiepileptic drug, to prevent benzodiazepine withdrawal in an animal model sensitive to the anxiogenic effect resulting from drug cessation. The effects of levetiracetam (17 and 54 mg/kg) given intraperitoneally (i.p.) were determined on anxiety induced in female NMRI mice by withdrawal from 21 days of chronic administration of chlordiazepoxide. Administration of chlordiazepoxide was i.p. twice daily, in increments of 2 mg/kg, from 10 up to 40 mg/kg. Anxiety was evaluated using an elevated plus-maze test 24-h after chlordiazepoxide withdrawal. Discontinuation of chronic chlordiazepoxide induced a significant anxiogenic profile in the plus-maze test mainly characterised by a decrease in open arm exploration. This effect was dose dependently prevented by administration of levetiracetam during the withdrawal period. The highest dose tested (54 mg/kg) induced statistically significant effects on all variables recorded but had no effect upon plus-maze exploration in normal mice. This suggests that the observed effects are dependent upon the level of stress or anxiety of the animals. These results support potential efficacy of levetiracetam in the benzodiazepine withdrawal syndrome. PMID- 11937099 TI - Repeated administration of the neurotensin analogue NT69L induces tolerance to its suppressant effect on conditioned avoidance behaviour. AB - Although acute neurotensin receptor stimulation exerts diverse behavioural effects that resemble those seen after administration of antipsychotic drugs, data on effects after repeated exposure to neurotensin receptor agonism is relatively sparse. Here, we demonstrate that repeated administration of the novel neurotensin-(8-13) analogue NT69L [(N-methyl-Arg), Lys, Pro, L-neo-Trp, tert-Leu, Leu] induce tolerance to its suppressant effect on conditioned avoidance behaviour in rats, a predictive assay for antipsychotic activity. In contrast, the inhibitory effect of haloperidol on this behaviour was sustained despite repeated administration of this classical antipsychotic drug. These findings indicate that repeated exposure to neurotensin receptor stimulation induces tolerance to the antipsychotic-like effects of neurotensin receptor agonists. PMID- 11937100 TI - Autoinhibitory function of the sympathetic prejunctional neuropeptide Y Y(2) receptor evidenced by BIIE0246. AB - The significance of neuropeptide Y Y(2) receptors in sympathetic nonadrenergic transmission was investigated using the novel selective antagonist BIIE0246 ((S) N2-[[1-[2-[4-[(R,S)-5,11-dihydro-6(6h)-oxodibenz[b,e]azepin-11-yl]-1-piperazinyl] 2-oxoethyl]cyclopentyl]acetyl]-N-[2-[1,2-dihydro-3,5 (4H)-dioxo-1,2-diphenyl-3H 1,2,4-triazol-4-yl]ethyl]-argininamide). In anaesthetized pigs pretreated with reserpine, and after transection of sympathetic nerves (depleted of noradrenaline), electrical stimulation of renal and splanchnic sympathetic nerves evoked vasoconstriction in, and overflow of neuropeptide Y-like immunoreactivity from, kidney and spleen, respectively. In the presence of BIIE0246, the nerve evoked overflows of neuropeptide Y-like immunoreactivity were markedly increased and the splenic vasoconstrictor response prolonged. In addition, BIIE0246 caused splenic vasodilatation per se in this model where basal levels of circulating neuropeptide Y exceed 40 pM. It is concluded that endogenous neurogenical neuropeptide Y regulates its own release via activation of sympathetic prejunctional inhibitory neuropeptide Y Y(2) receptors in both spleen and kidney in the reserpinized pig. Moreover, when circulating levels of neuropeptide Y are moderately increased, activation of neuropeptide Y Y(2) receptors seems to contribute to basal splenic vascular tone. PMID- 11937101 TI - Experimental diabetes induces hyperreactivity of rabbit renal artery to 5 hydroxytryptamine. AB - The influence of diabetes on the response of isolated rabbit renal arteries to 5 hydroxytryptamine (5-HT) was examined. 5-HT induced a concentration-related contraction that was higher in arteries from diabetic rabbits than in arteries from control rabbits. Endothelium removal did not significantly modify 5-HT contractions in arteries from control rabbits but enhanced the response to 5-HT in arteries from diabetic rabbits. Incubation with N(G)-nitro-L-arginine (L-NA) enhanced contractions to 5-HT in arteries from control and diabetic rabbits. In arteries with endothelium, this L-NA enhancement was lower in diabetic rabbits than in control rabbits. In arteries without endothelium, incubation with L-NA enhanced the maximal contractions to 5-HT in control rabbits but did not in diabetic rabbits. Indomethacin inhibited 5-HT-induced contraction of arteries from control rabbits and enhanced the maximal contraction to 5-HT of arteries from diabetic rabbits. In summary, diabetes enhances contractile response of rabbit renal artery to 5-HT. In control animals, this response is regulated by both endothelial and non-endothelial (neuronal) nitric oxide (NO) and by a vasoconstrictor prostanoid. Diabetes impairs the release of non-endothelial NO and the vasoconstrictor prostanoid. PMID- 11937102 TI - Reversal of tolerance to nitroglycerin with N-acetylcysteine or captopril: a role of calcitonin gene-related peptide. AB - Previous studies have shown that the development of tolerance to nitroglycerin is related to a decrease in the release of endogenous calcitonin gene-related peptide (CGRP). In the present study, we explored whether endogenous CGRP is involved in reversal of tolerance to nitroglycerin with N-acetylcysteine or captopril in rats in vivo and vitro. Tolerance was induced by exposure to nitroglycerin (4.4 x 10(-6) M) for 10 min in vitro or by pretreatment with nitroglycerin (10 mg/kg, s.c.) three times a day for 8 days in vivo. Nitroglycerin (3 x 10(-9)-10(-6) M) caused a concentration-dependent relaxation in the isolated rat thoracic aorta, an effect that was reduced by CGRP-(8-37) (3 x 10(-7) M) or capsaicin (3 x 10(-7) M). Preincubation with nitroglycerin for 10 min significantly decreased its vasodilation, which was restored in the presence of N-acetylcysteine (10(-5) M) or captopril (10(-5) M). Nitroglycerin (150 microg/kg, i.v.) produced a depressor effect and an increase in concentrations of nitric oxide and CGRP, and the effects of nitroglycerin disappeared after pretreatment with nitroglycerin for 8 days. However, tolerance to nitroglycerin in vivo also was partially restored in the presence of N-acetylcysteine or captopril. The present results suggest that reversal of tolerance to nitroglycerin with N-acetylcysteine or captopril is related to the increased release of CGRP in the rat. PMID- 11937103 TI - AG-041R, a cholecystokinin-B/gastrin receptor antagonist, stimulates the repair of osteochondral defect in rabbit model. AB - A newly synthesized compound (AG-041R), 3R-1-(2,2Diethoxyethyl)-((4methylphenyl) amino-carbonyl methyl)-3-((4methylphenyl) ureido-indoline-2-one), is a cholecystokinin-B/gastrin receptor antagonist which has stimulatory effects on the matrix synthesis of chondrocytes in vitro. In this study, we examined the effect of AG-041R on the repair of osteochondral defects (cylindrical, 4 mm diameter) in the patellar groove of the rabbit knee joint. At the time of operation, 100 microl of 1 microM of AG-041R was administered, followed by 200 microl with an osmotic pump for 14 days. Histological and biochemical evaluations were performed at 12 and 24 weeks after surgery. The histological score of the AG 041R-treated group, the quantity of glycosaminoglycan and the ratio of chondroitin sulfate in the AG-041R-treated tissue were significantly higher than in the untreated group. Moreover, the degeneration of cartilage around the defect was suppressed in the AG-041R-treated group. These findings suggest that AG-041R is effective for the repair of osteochondral defects. PMID- 11937104 TI - Cholesterol lowering effects of a choleretic phloracetophenone in hypercholesterolemic hamsters. AB - The plasma cholesterol-lowering effect and mechanism thereof of a choleretic phloracetophenone or 2,4,6-trihydroxyacetophenone (THA) were investigated in hypercholesterolemic male hamsters. Intragastric administration of THA (300-600 micromol/kg) twice a day for 7 days to these animals caused a dose- and time dependent decrease in both plasma cholesterol and triglyceride levels. THA at a dose of 400 micromol/kg reduced the cholesterol and triglyceride levels in plasma to 52% and 25% of the level in corresponding cholesterol-fed controls, respectively, with decreases in both plasma very low density lipoprotein and low density lipoprotein cholesterol but not in high density lipoprotein cholesterol. THA did not significantly alter total hepatic cholesterol content but significantly increased the excretion of both bile acids and cholesterol into the intestinal lumen for elimination. Corresponding to the increase in bile acid excretion, THA caused a seven-fold increase in hepatic cholesterol 7alpha hydroxylase activity. These results suggest that THA exerts its cholesterol lowering effect by increasing hepatic cholesterol 7alpha-hydroxylase activity which increases hepatic conversion of cholesterol to bile acid for disposal via biliary secretion. This compound may have a potential for future development as a therapeutic agent for lowering lipids in hypercholesterolemic patients. PMID- 11937105 TI - Role of tachykinins in sephadex-induced airway hyperreactivity and inflammation in guinea pigs. AB - We have studied the effect of selective tachykinin NK(1) and NK(2) receptor antagonists on airway hyperreactivity to acetylcholine and increase of inflammatory cells on bronchoalveolar lavage fluid induced by sephadex beads (20 mg/kg, i.v.) in guinea pigs. Airway hyperreactivity was assessed by measuring the increase of bronchial insufflation pressure to acetylcholine (0.01-30 micromol/kg, i.v.) at 3 h (early phase) and 24 h (late phase) after sephadex administration. An increase in inflammatory cells in bronchoalveolar lavage fluid (eosinophils and macrophages) was detected at 24 h (from 11.6 x 10(6) to 49.3 x 10(6) cells) but not at 3 h from sephadex administration. Neurokinin A and substance P levels in bronchoalveolar lavage fluid showed a significant increase at 24 h (from 31.7+/-11.6 to 561+/-231 pg/ml and from 5.9+/-2.6 to 29.3+/-4.1 pg/ml for neurokinin A and substance P, respectively). At this time point, the tachykinin in bronchoalveolar lavage cellular content was depleted from 232+/-43 to 21+/-20 pg/sample and from 56.6+/-6.7 to 2+/-2 pg/sample for neurokinin A and substance P, respectively. Capsaicin pretreatment abolished the early but not the late phase of airway hyperreactivity induced by sephadex without modifying bronchoalveolar lavage total cells number and bronchoalveolar lavage levels of neurokinin A and substance P. Administration of the tachykinin NK(2) (nepadutant) and/or the NK(1) receptor antagonist (MEN 11467 or (1R,2S)-2-N[1(H)indol-3-yl carbonyl]-1-N[N-(p-tolylacetyl)-N-(methyl)-D-3(2 naphthyl)alanyl)diaminocyclohexane)), 5 min before sephadex, prevented the early phase of airway hyperreactivity to acetylcholine but only nepadutant prevented the late phase. Nepadutant was able to abolish the early phase of airway hyperreactivity if given after sephadex administration and reduced by about 50% the increase of cell number in bronchoalveolar lavage fluid during the late phase, without affecting the levels of neurokinin A and substance P. These findings indicate an involvement of endogenous tachykinins in the genesis of airway hyperreactivity in a guinea-pig model of non-allergic asthma. Early airway hyperreactivity apparently involves release of tachykinins from capsaicin sensitive afferent nerves acting via tachykinin NK(1)/NK(2) receptors. Late airway hyperreactivity involves tachykinins acting via tachykinin NK(2) receptors: inflammatory cells activated/recruited in response to sephadex challenge appear a likely source of tachykinins involved in the late phase of the response. PMID- 11937106 TI - No involvement of interleukin-5 or eosinophils in experimental allergic rhinitis in guinea pigs. AB - The aim of this study is to evaluate whether nasal airway eosinophilia is a true pathogenetic component of allergic rhinitis. We investigated the effects of TRFK5, an anti-interleukin-5 antibody, not only on leukocyte mobilization from the bone marrow, but also on the development of nasal symptoms and hyperresponsiveness in a guinea pig model of allergic rhinitis. Intranasally sensitized animals were repetitively challenged by exposure to Japanese cedar pollen as antigen. TRFK5 (100 microg/kg, i.p.) given 12 h before the final antigen challenge selectively prevented the antigen-induced eosinophilia in blood and the nasal airway, and suppressed the corresponding decrease in the number of cells in bone marrow; however, it failed to inhibit the immediate development of sneezing, early and late nasal blockage responses, goblet cell degranulation and nasal hyperresponsiveness to histamine. Furthermore, TRFK5 did not significantly affect the production of thromboxane A(2) and cysteinyl leukotrienes in the nasal airway during the late response. These results strongly suggest that while interleukin-5 is essential for eosinophil migration from the bone marrow to the nasal airway, neither interleukin-5 nor eosinophils are required for the development of the nasal symptoms and nasal hyperresponsiveness of allergic rhinitis. PMID- 11937107 TI - Constitutively active melatonin MT(1) receptors in male rat caudal arteries. AB - This study assessed the state of melatonin MT(1) receptor coupling in sections of male rat caudal arteries by [35S]GTPgammaS binding autoradiography. The melatonin MT(1) receptor inverse agonist 4-phenyl-2-propionamidotetraline (4P-PDOT) (0.1-1 microM) significantly decreased [35S]GTPgammaS binding compared to basal, strongly suggesting the presence of constitutively active receptors. Formation of constitutively active receptors during subjective day, when the levels of melatonin are low, may be a physiological mechanism by which the organism maintains vascular tone. PMID- 11937108 TI - Nitric oxide inhibits RhoA/Rho-kinase signaling to cause penile erection. AB - The RhoA/Rho-kinase pathway mediates vasoconstriction in the cavernosal circulation. Inhibition of this pathway leads to penile erection in the in vivo rat model. These studies examined the hypothesis that nitric oxide (NO) inhibits RhoA/Rho-kinase signaling as part of normal erection. The results show that NO causes increased intracavernosal pressure and that this response is potentiated by prior treatment with a threshold dose of the Rho-kinase inhibitor, (+)-(R) trans-4-(1-Aminoethyl)-N-(4-pyridyl) cyclohexanecarboxamide dihydrochloride, monohydrate (Y-27632). These results support the hypothesis that NO inhibits Rho kinase-induced cavernosal vasoconstriction during erection. PMID- 11937109 TI - Gene therapy and reproductive medicine. AB - OBJECTIVE: To review the literature on the principles of gene therapy and its potential application in reproductive medicine. DESIGN: Literature review. SETTING: Gene therapy involves transfer of genetic material to target cells using a delivery system, or vector. Attention has primarily focused on viral vectors. Significant problems remain to be overcome including low efficacy of gene transfer, the transient expression of some vectors, safety issues with modified adenoviruses and retroviruses, and ethical concerns. If these issues can be resolved, gene therapy will be applicable to an increasing spectrum of single and multiple gene disorders, as the Human Genome Project data are analyzed, and the genetic component of human disease becomes better understood. Gynecologic gene therapy has advanced to human clinical trials for ovarian carcinoma, and shows potential for the treatment of uterine leiomyomata. Obstetric applications of gene therapy, including fetal gene therapy, remain more distant goals. CONCLUSION(S): Concerns about the safety of human gene therapy research are being actively addressed, and remarkable progress in improving DNA transfer has been made. The first treatment success for a genetic disease (severe combined immunodeficiency disease) has been achieved, and ongoing research efforts will eventually yield clinical applications in many spheres of reproductive medicine. PMID- 11937110 TI - What is an embryo? PMID- 11937111 TI - Female androgen insufficiency: the Princeton consensus statement on definition, classification, and assessment. AB - OBJECTIVE: To evaluate the evidence for and against androgen insufficiency as a cause of sexual and other health-related problems in women and to make recommendations regarding definition, diagnosis, and assessment of androgen deficiency states in women. DESIGN: Evaluation of peer-review literature and consensus conference of international experts. SETTING: Multinational conference in the United States. PATIENT(S): Premenopausal and postmenopausal women with androgen deficiency. INTERVENTION(S): Evaluation of peer-review literature and development of consensus panel guidelines. RESULT(S): The term "female androgen insufficiency" was defined as consisting of a pattern of clinical symptoms in the presence of decreased bioavailable T and normal estrogen status. Currently available assays were found to be lacking in sensitivity and reliability at the lower ranges, and the need for an equilibrium dialysis measure was strongly emphasized. Causes of androgen insufficiency in women were classified as ovarian, adrenal, hypothalamic-pituitary, drug-related, and idiopathic. A simplified management algorithm and clinical guidelines were proposed to assist clinicians in diagnosis and assessment. Androgen replacement is currently available in several forms, although none has been approved for treatment of sexual dysfunction or other common symptoms of female androgen insufficiency. Potential risks associated with treatment were identified, and the need for informed consent and careful monitoring was noted. Finally, the panel identified key goals and priorities for future research. CONCLUSION(S): A new definition of androgen insufficiency in women has been proposed along with consensus-based guidelines for clinical assessment and diagnosis. A simplified management algorithm for women with low androgen in the presence of clinical symptoms and normal estrogen status has also been proposed. PMID- 11937113 TI - Pregnancies following use of metformin for ovulation induction in patients with polycystic ovary syndrome. AB - OBJECTIVE: To assess pregnancy outcome in anovulatory infertility patients diagnosed with polycystic ovary syndrome (PCOS) who were treated with metformin. DESIGN: Case series. SETTING: Outpatient. PATIENT(S): Anovulatory patients (n = 48) with a diagnosis of PCOS based on clinical, diagnostic, and laboratory evaluations were enrolled in the study over a 15-month period. INTERVENTION(S): Metformin was started at 500 mg b.i.d. for 6 weeks and then increased to 500 mg t.i.d. if no ovulation occurred. Clomiphene citrate (CC; 50 mg) was added if no ovulatory response occurred after 6 weeks. MAIN OUTCOME MEASURE(S): Resumption of menses, presumptive ovulation, and pregnancy. RESULT(S): Nineteen of 48 (40%) patients resumed spontaneous menses following treatment and showed presumptive evidence of ovulation with metformin alone; 15/48 (31%) required CC (50 mg) in conjunction with metformin therapy, and 10 of these 15 (67%) had evidence of ovulation; 20/48 (42%) conceived with a median time to conception of 3 months, and 7 of these 20 (35%) had spontaneous abortions (SAB); 19/48 (40%) had gastrointestinal-related side effects, and 5 of 48 patients (10%) had to decrease the dosage of metformin. Only 1 patient discontinued therapy. CONCLUSION(S): Metformin alone in patients with PCOS results in a substantial number of pregnancies, with 69% (20/29) of those who ovulated conceiving in less than 6 months. PMID- 11937112 TI - Insulin gene variable number of tandem repeats regulatory polymorphism is not associated with hyperandrogenism in Spanish women. AB - OBJECTIVE: To determine if the insulin gene variable number of tandem repeats (VNTR) regulatory polymorphism is associated with hyperandrogenism in a population of Spanish women. DESIGN: Controlled clinical study. SETTING: Tertiary institutional hospital. PATIENT(S): Ninety-six hyperandrogenic patients and 38 healthy control women. INTERVENTION(S): Whole blood and serum samples were collected during the follicular phase of the menstrual cycle. MAIN OUTCOME MEASURE(S): Insulin gene VNTR regulatory polymorphism genotypes (classes I/I, I/III, and III/III alleles) and serum androgen levels. Insulin resistance was estimated from fasting glucose and insulin levels by using the homeostatic model assessment. RESULT(S): The frequencies of VNTR genotypes were 45.5%, 43.3%, and 11.2% for I/I, I/III, and III/III alleles considering patients and controls as a whole. These frequencies were not statistically different in controls (47.4%, 34.2%, and 18.4%) and in patients (44.8%, 46.9%, and 8.3%). CONCLUSION(S): Hyperandrogenism and the insulin gene VNTR regulatory polymorphism are not associated in Spanish women. PMID- 11937114 TI - Epidemiologic and etiologic aspects of hirsutism in Kashmiri women in the Indian subcontinent. AB - OBJECTIVE: To assess the prevalence of hirsutism and study its etiology in the Kashmir Valley of the Indian subcontinent. DESIGN: Prospective random sample of the general population attending outpatient medical clinics and prospective evaluation of hirsute patients referred to our endocrinology clinic. SETTING: A tertiary care hospital and district-level primary care hospitals. PATIENT(S): Five thousand women attending various hospitals for reasons unrelated to hirsutism and 150 consecutive women referred for hirsutism. INTERVENTION(S): Assessment of body hair as per the Ferriman and Gallwey scoring system and an investigative protocol including detailed clinical assessment with endocrinologic workup including estimations of gonadotropins, PRL, T, and 17-hydroxyprogesterone and abdominopelvic ultrasound. MAIN OUTCOME MEASURE(S): Presence and cause of hirsutism. RESULT(S): Of 4,780 adult women for whom adequate data were available, 504 (10.5%) had hirsutism, among whom 484 (10.1%) had mild (score of 6-9) and 20 (0.4%) had moderate hirsutism (score of 10-14). The etiology of hirsutism revealed idiopathic hirsutism in 38.7%, polycystic ovary syndrome (PCOS) in 37.3%, postmenopausal state in 9.2%, adrenal tumors in 2.1%, congenital adrenal hyperplasia in 1.4%, and drug-induced hirsutism in 0.7%. The cause remained undetermined in 10.6% of patients for whom the available information was not adequate. CONCLUSION(S): Hirsutism is as common a problem in the Kashmir Valley (India) as elsewhere in the world. Idiopathic hirsutism (38.7%), PCOS (37.3%), and postmenopausal state (9.2%) are common causes of hirsutism. Late-onset congenital adrenal hyperplasia is a relatively uncommon cause of hirsutism in the Kashmir Valley. PMID- 11937115 TI - Aberrant expression of intercellular adhesion molecule-1 and killer inhibitory receptors induces immune tolerance in women with pelvic endometriosis. AB - OBJECTIVE: To investigate host immunologic response to endometriosis in terms of intercellular adhesion molecule (ICAM)-1 expression by macrophages and killer cell inhibitory receptor (KIR) expression by natural killer (NK) cells. DESIGN: Case-control study of immunologic markers. SETTING: University hospital. PATIENT(S): Twenty-eight Japanese women with endometriosis. Control subjects were 26 women without endometriosis. Diagnoses were made at laparoscopy. INTERVENTION(S): Venipuncture and laparoscopic peritoneal fluid collection. MAIN OUTCOME MEASURE(S): ICAM-1 expression by macrophages and KIR expression by NK cells, measured by flow cytometry. RESULT(S): In women with endometriosis, expression of ICAM-1 by peritoneal macrophages was significantly lower and expression of KIR by NK cells in peritoneal fluid and peripheral blood was significantly higher than in control subjects. CONCLUSION(S): Properties of macrophages and NK cells in women with endometriosis promote immunotolerance to implanted tissue in the peritoneal environment. Increased KIR(+)NK cells in peripheral blood may represent a risk factor for endometriosis. PMID- 11937116 TI - Dienogest is as effective as triptorelin in the treatment of endometriosis after laparoscopic surgery: results of a prospective, multicenter, randomized study. AB - OBJECTIVE: To compare the efficacy of Dienogest versus Decapeptyl at 3.75 mg as consolidation therapy for surgery in the treatment of endometriosis. DESIGN: Multicenter, open, randomized, parallel-group clinical trial. SETTING: Volunteer patients in an academic research environment. PATIENT(S): Women with grade 2, 3, and 4 (5 years ago experienced a marginal increase in volume of menstrual flow compared with women with no tubal ligation history. CONCLUSION(S): We found no significant change in menstrual cycle characteristics or hormone levels in women with or without a history of tubal ligation. However, tubal ligation may have a modest effect on the change in menstrual flow volume over time among parous women with a history of cesarean section. PMID- 11937128 TI - Adrenal progestogen and androgen production in 21-hydroxylase-deficient nonclassic adrenal hyperplasia is partially independent of adrenocorticotropic hormone stimulation. AB - OBJECTIVE: To test the hypothesis that adrenal steroidogenesis in nonclassic adrenal hyperplasia (NCAH) patients is, at least in part, independent of adrenocorticotropic hormone (ACTH) control. DESIGN: Prospective controlled clinical study. SETTING: Patients and healthy volunteers in an academic research environment. PATIENT(S): Four patients with 21-hydroxylase (21-OH) deficient NCAH and four healthy control women. INTERVENTION(S): Patients received the long acting gonadotropin-releasing hormone analog (GnRH-a) leuprolide acetate (3.75 mg/month IM) on weeks 0 and 4; and dexamethasone (DEX) in weekly incremented doses (0.25 mg/day, 0.50 mg/day, 1.0 mg/day, and 1.5 mg/day), beginning on weeks 4, 5, 6, and 7, respectively. MAIN OUTCOME MEASURE(S): The levels of 17 hydroxyprogesterone (17-HP), progesterone (P4), androstenedione (A4), dehydroepiandrosterone sulfate (DHS), and cortisol (F) were measured at the beginning of weeks 0, 4, 5, 6, 7, and at the end of the study (beginning of week 8). RESULT(S): Patients and controls had a similar median age and body mass index (BMI). There were no significant decreases in the median levels of the studied hormones following 4 weeks of treatment with GnRH-a only, in either NCAH patients or controls. Analysis of individual hormonal values demonstrated that by the end of the study (after DEX of 1.5 mg/day during a week) only 2 of 4, 0 of 4, 3 of 4 and 3 of 4 NCAH patients had 17-HP, P4, A4, and DHS levels within the range of control values, respectively. CONCLUSION(S): Ovarian and incremental adrenal suppression did not fully suppress progestogen and androgen production in all of the study patients with 21-OH-deficient NCAH, suggesting that their production was partially independent of ACTH stimulation. Potentially in these patients subtle degrees of adrenocortical hyperplasia and/or abnormal enzymatic kinetics are responsible for the nonsupressibility. PMID- 11937130 TI - Efficacy of methotrexate treatment in extrauterine pregnancies defined by stable or increasing human chorionic gonadotropin concentrations. AB - OBJECTIVE: To evaluate the efficacy of methotrexate treatment in selected cases of extrauterine pregnancy (EUP) defined by stable or increasing hCG concentration. DESIGN: Prospective cohort study. SETTING: Gynecology department of the Lis Maternity Hospital. PATIENT(S): Fifty women with EUP diagnosed whenever an intrauterine gestational sac was not seen on transvaginal ultrasonography. INTERVENTION(S): Women received IM methotrexate at a dose of 50 mg/m(2) of body surface area. Failure of hCG levels to fall by >/=15% during any successive week resulted in repeated administration of methotrexate. Surgical intervention was performed for presumed tubal rupture. MAIN OUTCOME MEASURE(S): Serial hCG measurement was performed weekly until hCG concentration reached 15 mIU/mL. Success was defined as the achievement of hCG concentration of 25 mIU/mL without surgical intervention. RESULT(S): Forty-four women (88%) were successfully treated. The mean time from first methotrexate injection to success was 34 +/- 2.4 days. Women treated successfully and unsuccessfully differed significantly only with regard to serum hCG levels of 1,876 +/- 243 and 3,489 +/- 376 mIU/mL, respectively. When the initial hCG levels were lower or higher than 2,000 IU/L, the success rate was 97% and 74%, respectively (significant by Fisher's exact test). CONCLUSION(S): When methotrexate treatment is administrated in a selected group of EUP defined by stable or increasing hCG, it may fail more frequently (26%) when initial hCG levels are >2,000 mIU/mL. PMID- 11937131 TI - Postoperative infection and surgical hysteroscopy. AB - OBJECTIVE: To evaluate the risk of infection after surgical hysteroscopy. DESIGN: Prospective observational study. SETTING: University hospital. PATIENT(S): One thousand nine hundred fifty-two patients requiring operative hysteroscopy during a 10-year period from January 1990 to January 2000. INTERVENTION(S): Two thousand one hundred sixteen operative hysteroscopies were performed: 782 fibroma resections, 422 polyp resections, 623 endometrectomies, 90 uterine septa sections, and 199 lyses of synechiae. MAIN OUTCOME MEASURE(S): Postoperative infectious complications. RESULT(S): Thirty (1.42%) infections occurred. There were 18 (0.85%) cases of endometritis and 12 urinary tract infections. No other severe infectious complications were reported. The risk for early-onset endometritis was higher after lysis of synechiae compared with endometrectomy, fibroma, or polyp resections. However, the risk for early-onset endometritis was similar for endometrectomy, septa, fibroma, or polyp resections. CONCLUSION(S): Infectious risk following surgical hysteroscopy is low. No major infectious complications occurred. Risk for early-onset endometritis was higher after lysis of synechiae compared with other procedures. PMID- 11937132 TI - Transvaginal ultrasound-guided embryo transfer improves outcome in patients with previous failed in vitro fertilization cycles. AB - OBJECTIVE: To determine the effect of transvaginal ultrasound-guided ET in IVF cycles performed on patients who had previously failed to conceive from IVF and compare the results to previous cycles where ultrasound guidance was not used. DESIGN: Retrospective clinical study. SETTING: Private practice IVF program. PATIENT(S): One hundred twenty-nine women undergoing consecutive cycles of IVF where fresh embryos were transferred. INTERVENTION(S): Transvaginal ultrasound guidance was used during transfer of embryos. MAIN OUTCOME MEASURE(S): Patient age, number of ampules of gonadotropin used, maximum E(2) level, number of oocytes retrieved, number of two pronuclei embryos obtained, number of embryos transferred, mean embryo score, implantation and pregnancy rate. RESULT(S): There was no difference in any of the clinical parameters measured in IVF cycles resulting in pregnancy when transvaginal ultrasound-guided ET was used compared to the failed cycles when there was no ultrasound guidance. Of the patients who previously had failed IVF cycles and subsequently had IVF cycles with ultrasound guidance, those who became pregnant had higher mean embryo scores than those who did not become pregnant. Overall implantation and pregnancy rates were higher during the study period when transvaginal ultrasound guidance was used than in the previous 3 years when it was not used. CONCLUSION(S): Transvaginal ultrasound guided ET may be responsible for successful IVF cycles in patients who had previously failed to conceive when embryos were transferred by the clinical touch method. Transvaginal ultrasound guidance may also be responsible for an overall increase in embryo implantation and pregnancy compared to the use of the clinical touch method. PMID- 11937133 TI - Aromatase inhibition improves ovarian response to follicle-stimulating hormone in poor responders. AB - OBJECTIVE: To examine the use of the aromatase inhibitor letrozole with FSH for ovarian stimulation in poor responders undergoing ovarian superovulation and IUI. DESIGN: Observational cohort study as a prospective clinical trial in patients with unexplained infertility and a low response to ovarian stimulation with FSH. SETTING: Two tertiary referral infertility clinics associated with the Reproductive Sciences Division, University of Toronto. PATIENT(S): Twelve patients with unexplained infertility undergoing IUI who received FSH alone in 25 prior cycles with poor response (less than three dominant follicles). INTERVENTION(S): Patients were offered letrozole, 2.5 mg/day from day 3-7 of the menstrual cycle with FSH (50-225 IU/day) starting on day 5-7. hCG (10,000 IU) was given when two leading follicles were >/=2 cm followed by IUI. MAIN OUTCOME MEASURE(S): Number of mature follicles (>1.8 cm), FSH dose, endometrial thickness, and pregnancy rate. RESULT(S): Improved response to FSH stimulation with letrozole co-treatment was evidenced by the significantly lower FSH dose associated with significantly higher number of mature follicles. During letrozole plus FSH stimulation cycles, clinical pregnancy was achieved in three cycles (21%). CONCLUSION(S): In this preliminary report, we demonstrate a potential benefit of aromatase inhibition for improving ovarian response to FSH in poor responders. PMID- 11937134 TI - Increased endometrial thickness on the day of human chorionic gonadotropin injection does not adversely affect pregnancy or implantation rates following in vitro fertilization-embryo transfer. AB - OBJECTIVES: To investigate the controversy whether an increased endometrial thickness has an effect on pregnancy, implantation, or abortion rates in in vitro fertilization-embryo transfer (IVF-ET) cycles. DESIGN: Retrospective analysis. SETTING: A university-based IVF center. PATIENT(S): Five hundred seventy women under the age of 40. INTERVENTION(S): Measurements of endometrial thickness on day of human chorionic gonadotropin (hCG) administration. Cycles were compared by endometrial thickness of 14 mm in 60 women. MAIN OUTCOME MEASURE(S): Implantation, pregnancy, and abortion rates. RESULT(S): Implantation, pregnancy, and abortion rates were similar in each group. In cycles where the endometrial thickness was 14 mm. CONCLUSION(S): No adverse effects of a thickened endometrium were demonstrated on implantation, pregnancy, or abortion rates in the first IVF-ET cycle. These findings fail to corroborate with those of Weissman et al. and support those of Yakin et al. PMID- 11937135 TI - Apoptosis and expression of apoptosis-related genes in the mouse testis following heat exposure. AB - OBJECTIVE: To investigate molecular mechanisms of germ cell apoptosis induced by heat exposure in mice. DESIGN: Controlled laboratory study. SETTING: Departments of Urology and Biochemistry, Yamagata University School of Medicine, Yamagata, Japan. ANIMAL(S): Forty-four male B6D2F1 mice. INTERVENTION(S): Heat exposure, 43 degrees C for 15 minutes. MAIN OUTCOME MEASURE(S): Testicular germ cell apoptosis (percentages of apoptotic tubules and apoptotic cells) was assessed by using DNA nick-end labeling, and expression of Bcl-2 family, Fas-FasL system, and p53 was evaluated by using Western analysis. RESULT(S): Bilateral testicular weights decreased significantly from 3 days after heat exposure. Percentages of apoptotic tubules and apoptotic germ cells increased significantly from 1 day after heat exposure. There were no significant changes in the levels of Bcl-xl, Bad, and Bax after heat exposure. However, Bcl-2 expression level decreased significantly 7 days after heat exposure. In contrast, the expression level of Fas and p53 increased significantly from 1 day to 3 days after heat exposure, respectively. Expression level of FasL elevated significantly at days 1 and 2 but declined from day 3. CONCLUSION(S): Germ cell apoptosis induced by heat exposure is mainly mediated by the Fas-FasL system. PMID- 11937136 TI - Rhesus offspring produced by intracytoplasmic injection of testicular sperm and elongated spermatids. AB - OBJECTIVE: To establish pregnancies in rhesus monkeys using testicular sperm and elongated spermatids injected into oocytes. DESIGN: Comparative animal study. SETTING: Regional Primate Research Center. ANIMAL(S): Prime, fertile rhesus monkeys. INTERVENTION(S): Oocytes collected by laparoscopy from gonadotropin stimulated female rhesus monkeys were injected with testicular sperm or elongated spermatids obtained from the testis of males. Cleavage stage embryos were transferred to surrogate females. MAIN OUTCOME MEASURE(S): Fertilization, embryo cleavage, and the establishment of pregnancies. Fertilization failures were fixed and processed for the detection of microtubules and chromatin configurations. RESULT(S): Fertilization, assessed by the presence of two pronuclei within 15 hours after injection, was 60% for intracytoplasmic sperm injection with testicular sperm and 47% for elongated spermatid injection. Fertilized zygotes co cultured in Connaughts Medical Research Labs (CMRL) medium on a Buffalo Rat Liver cell monolayer resulted in hatched blastocysts after testicular sperm extraction intracytoplasmic sperm injection and elongated spermatids. Embryos transferred at the 4- to 8-cell stage gave rise to three pregnancies: 2/3 from testicular sperm and 1/1 from an elongated spermatid. Three healthy infants were delivered by cesarean. Oocytes that failed to fertilize typically remained arrested in metaphase of meiosis. CONCLUSION(S): Testicular sperm and elongated spermatids can be used for fertilization in the rhesus monkey resulting in live births. PMID- 11937137 TI - Aminopeptidases in the gonads of male and female rats. AB - OBJECTIVE: To assess the relative importance of soluble (Sol) and membrane-bound (M-B) components of aminopeptidase (AP) activities in local functions of male and female gonads. DESIGN: Observational study. SETTING: University research. ANIMAL(S): Adult male and female Sprague-Dawley rats in the proestrous phase of the estrous cycle. INTERVENTION(S): Samples from the right testis and the whole right ovary were dissected after perfusion with saline. The Sol and M-B fractions were obtained from these samples. MAIN OUTCOME MEASURE(S): Fluorometric measurement of Sol and M-B AP activities using arylamide derivatives as substrates. RESULT(S): Highly significant differences between male and female gonads were observed. Sol AP activities showed a significant predominance in testes, whereas M-B AP activities were significantly higher in ovaries. CONCLUSION(S): There was a discrepancy in the distribution of Sol and M-B AP activities between male and female gonads, which may imply a direct participation of sex steroids in AP activities. These results may reflect the relative importance of these enzymes in the testis and ovary and should be taken into account in evaluating the functional role of peptides locally produced in gonads. PMID- 11937138 TI - Antiphospholid antibodies regulate the expression of trophoblast cell adhesion molecules. AB - OBJECTIVE: To examine the effect of antiphospholipid antibodies on trophoblast expression of adhesion molecules. DESIGN: Primary cytotrophoblast cell cultures. SETTING: Department of Obstetrics and Gynecology, Catholic University, Rome, Italy. PATIENT(S): Five normal pregnant women underwent uncomplicated vaginal delivery at 36 weeks of gestation. INTERVENTION(S): IgG antibodies were isolated from a patient with antiphospholipid syndrome and from a normal control subject, using protein-G Sepharose columns. Cytotrophoblast cells were dispersed in bicarbonate buffer containing trypsin and DNAse I. MAIN OUTCOME MEASURE(S): We investigated the effects of antiphospholipid antibodies on trophoblast adhesion molecules (alpha1 and alpha5 integrins, E and VE cadherins), both at the protein and mRNA levels. RESULT(S): The alpha1 and alpha5 integrins were present in trophoblast cells from 24 hours of culture. Treatment with IgG that were obtained from the patient with antiphospholipid syndrome significantly decreased alpha1 integrin and increased alpha5 integrin at both the mRNA and protein levels. Furthermore, IgG with antiphospholipid antibodies activities induced VE-cadherin down-regulation and the E-cadherin up-regulation at protein and mRNA levels compared with control IgG or untreated cells. CONCLUSION(S): The results suggest that the inadequate trophoblastic invasion, induced by antiphospholipid antibodies, can be the result of abnormal trophoblast adhesion molecules expression. PMID- 11937139 TI - Inhibition of mouse in vitro fertilization by an antibody against a unique 18 amino acid domain in the polysulfate-binding domain of proacrosin/acrosin. AB - OBJECTIVE: To determine the contribution of the polysulfate-binding domain (PSBD) of acrosin during sperm penetration. DESIGN: To inhibit the in vitro fertilization of mouse zona-intact oocytes by using a polyclonal antibody raised against an 18-amino acid peptide of proacrosin (anti-PSBD). SETTING: Unit of Reproduction and Development, Faculty of Biological Sciences, Pontifical Catholic University of Chile. PATIENT(S): None. INTERVENTION(S): A polyclonal antibody against the 43IFMYHNNRRYHTCGGILL(60) peptide was raised in New Zealand female rabbits. The specificity of the antibody was evaluated by an ELISA. Zona-intact mouse oocytes were coincubated with capacitated spermatozoa for 3 hours in the presence of 0.63 mg/mL of the antibody or preimmune serum. As a control, we used zona-free mouse oocytes under the same experimental conditions. MAIN OUTCOME MEASURE(S): We evaluated the fertilization rate of zona-intact and zona-free mouse oocytes by phase-contrast microscopy. An oocyte was considered fertilized when at least one decondensed sperm head was found within the egg cytoplasm. We evaluated 50-60 mouse oocytes in each group in three independent experiments. RESULT(S): The anti-PSBD antibody inhibited the fertilization of zona-intact, but not zona-free, mouse oocytes, by capacitated spermatozoa. In addition, the binding of the anti-PSBD to proacrosin/acrosin in a solid-phase assay was inhibited in the presence of polysulfates (fucoidan). CONCLUSION(S): The anti PSBD directed against the PSBD of proacrosin/acrosin inhibited the penetration of capacitated mouse spermatozoa through the zona pellucida. This antibody may be a useful tool to define the roles of the different domains of proacrosin/acrosin during gamete interaction. PMID- 11937140 TI - Effect of chilling on the organization of tubulin and chromosomes in rhesus monkey oocytes. AB - OBJECTIVE: To determine the effects of chilling on the organization and distribution of tubulin and chromosomes in rhesus monkey oocytes. DESIGN: Comparative laboratory study. SETTING: Academic research laboratory. ANIMAL(S): Eight adult female rhesus monkeys (Macaca mulatta) aged 6-16 years. INTERVENTION(S): A total of 171 oocytes retrieved from eight rhesus monkeys were separated into nine groups. One group of control oocytes was held at 37 degrees C during the experiment. Four groups of oocytes were rapidly cooled to 0 degrees C and held for 1, 5, 10, or 30 minutes and then fixed and stained. Four other groups of oocytes were cooled to 0 degrees C, held for 1, 5, 10, or 30 minutes, warmed and incubated at 37 degrees C for 60 minutes, and then fixed and stained. MAIN OUTCOME MEASURE(S): Organization of cytoskeleton and chromosomes. RESULT(S): Exposure of rhesus oocytes to 0 degrees C for only 1 minute resulted in complete depolymerization of tubulin. Incubation of chilled oocytes at 37 degrees C for 60 minutes caused partial restoration of tubulin, although most oocytes exhibited abnormal alignment of chromosomes and disorganized meiotic spindles. CONCLUSION(S): We conclude that rhesus monkey oocytes are extremely sensitive to chilling injury. Their successful cryopreservation may require rapid cooling to outpace this injury. PMID- 11937141 TI - A prospective randomized comparison of the Wallace catheter and the Cook Echo-Tip catheter for ultrasound-guided embryo transfer. AB - OBJECTIVES: To compare the performance of a new coaxial catheter system with an echo-dense tip (Cook Echo-Tip catheter) with a Wallace catheter during ultrasound guided ET. DESIGN: Prospective, randomized study. SETTING: Private academically affiliated infertility center. PATIENT(S): Two hundred fifty-one patients undergoing IVF-ET. INTERVENTION(S): The patients underwent ultrasound-guided ET by a single physician with a standardized technique alternately using one of the two catheters. MAIN OUTCOME MEASURE(S): Implantation rates, pregnancy rates, ease of transfer, visualization, and status of catheter tip. RESULT(S): The echo-dense tip of the Cook Echo-Tip catheter was consistently seen with ultrasound guidance, minimizing the need for catheter movement to identify the tip. Implantation rate (30% vs. 35%), clinical pregnancy rate (57% vs. 55%), and ongoing pregnancy rate (49% vs. 47%) were similar in both groups. CONCLUSION(S): The Cook Echo-Tip catheter with its echogenic tip simplifies ultrasound-guided ET, but pregnancy success rates are similar to those obtained when a Wallace catheter is used. PMID- 11937142 TI - Uterocervicoplasty with a bladder mucosa layer for the treatment of complete cervical agenesis. AB - OBJECTIVE: To create an endocervical canal in a patient with a complete cervical agenesis. DESIGN: Case report. SETTING: University hospital. PATIENT(S): A 12 year-old girl presented with lower abdominal pain. On examination, complete vaginal agenesis was noted, with a 2-cm vaginal dimple. A pelvic magnetic resonance imaging scan disclosed an hematometra and absence of the cervix and vagina. INTERVENTION(S): Initial surgical therapy consisted of a vaginoplasty with a sigmoid bowel segment and opening of the uterus by puncture and stenting. The cervical permeation failed, with immediate complete stenosis. A new attempt was made through a low sagittal hysterotomy by removing a central muscular cylinder and lining the channel with a free tubularized bladder mucosa graft. A stent was left in place. MAIN OUTCOME MEASURE(S): Hysteroscopy, hysterography, and clinical follow-up evaluation. RESULT(S): The cervical stent was removed after 5 months. A hysterography and hysteroscopy confirmed the permeability of the cervix, which was lined by a well-vascularized longitudinally folded mucosa. Regular menses had been noted for more than 3 years as of this report. CONCLUSION(S): Cervicoplasty with mucosal lining permits the creation of a patent cervical canal, even in the reputedly unfavorable forms of congenital cervical agenesis. PMID- 11937143 TI - Successful in vitro fertilization with anonymous donor oocytes in a patient with recurrent massive hemoperitoneum following spontaneous and induced ovulation. AB - OBJECTIVE: To report a case of severe von Willebrand's disease treated with in vitro fertilization using donor oocytes. DESIGN: Case report and literature review. SETTING: Private practice infertility center. PATIENT(S): Infertility patient with recurrent massive hemoperitoneum following spontaneous and induced ovulation. INTERVENTION(S): In vitro fertilization using donor oocytes. MAIN OUTCOME MEASURE(S): Successful term delivery. RESULTS(S): Delivery of healthy term singleton infant without maternal complication. CONCLUSION(S): The indications for in vitro fertilization using donor oocytes, while primarily for patients with medical/surgical menopause or genetic disorders, can be expanded to patients with contraindications for spontaneous or induced ovulation. PMID- 11937144 TI - Successful pregnancy in a woman with symptomatic fibroids who underwent laparoscopic bipolar coagulation of uterine vessels. AB - OBJECTIVE: To report a case of successful pregnancy after laparoscopic bipolar coagulation of uterine vessels (LBCUV). DESIGN: Case report. SETTING: University affiliated tertiary referral center. PATIENT(S): One woman, treated with LBCUV for symptomatic fibroids, who subsequently had a successful pregnancy. INTERVENTION(S): Laparoscopic bipolar coagulation of uterine arteries and anastomotic sites of uterine arteries with ovarian arteries. MAIN OUTCOME MEASURE(S): Patient evaluation by physical and ultrasound examinations. RESULT(S): Complete resolution of menorrhagia and dysmenorrhea was found after LBCUV. Reduction in fibroid size was seen by ultrasound. Two months later, the patient conceived a singleton pregnancy without the use of assisted reproductive technologies. The woman delivered by cesarean section. Neither myoma recurrence nor abnormality in uterine function was observed. CONCLUSION(S): Although fecundity- and pregnancy-related complications after LBCUV for managing uterine fibroids are still unclear, this first case report of successful pregnancy after LBCUV is promising. LBCUV might be a safe and effective alternative to myomectomy and hysterectomy. PMID- 11937145 TI - Postparacentesis bilateral massive vulvar edema in a patient with severe ovarian hyperstimulation syndrome. AB - OBJECTIVE: To report a case of bilateral massive vulvar edema following lower abdominal paracentesis in a patient with ovarian hyperstimulation syndrome. DESIGN: Case report. SETTING: University teaching hospital. PATIENT(S): A 32-year old woman with primary infertility. INTERVENTION(S): The patient underwent ovarian stimulation with leuprolide acetate, highly purified FSH, and hCG. Because of the development of severe ovarian hyperstimulation syndrome, bilateral paracentesis through the lower abdominal quadrants was performed. MAIN OUTCOME MEASURE(S): Treatment of ovarian hyperstimulation syndrome. RESULT(S): Development of bilateral massive vulvar edema 24 hours after lower abdominal paracentesis. CONCLUSION: This case report suggests that lower abdominal paracentesis could be the cause of vulvar edema development in ovarian hyperstimulation syndrome, probably due to a fistulous tract created between the peritoneal cavity and the subcutaneous tissues. PMID- 11937146 TI - Cytogenetic evaluation of human oocytes that failed to complete meiotic maturation in vitro. AB - OBJECTIVE: To determine the cause of infertility in a couple whose oocytes failed to mature in two consecutive fertility treatments. DESIGN: Case report. SETTING: University-based IVF program. PATIENT(S): A 32-year-old woman with unexplained infertility. INTERVENTION(S): Cytogenetic evaluation of oocytes that failed to reach meiotic metaphase II stage of maturation. MAIN OUTCOME MEASURE(S): Observation of oocyte maturity and chromosome composition after fixing and staining with Orcein stain. RESULT(S): Cytogenetic analysis revealed that the oocytes had successfully resumed meiosis. Germinal vesicle breakdown was also indicated, and chromosomes were at metaphase II stage of development. However, meiotic reduction of those chromosomes failed. CONCLUSION(S): Infertility in this couple seems to be attributed to the failure of the chromosomes to complete the reduction phase of metaphase II of meiosis. PMID- 11937147 TI - Benefit of vaginal sildenafil citrate in assisted reproduction therapy. PMID- 11937148 TI - Lateral distribution of recurrent ovarian endometriotic cysts. PMID- 11937149 TI - Contralateral corpus luteum in ectopic pregnancy: what does it tell us about ovum pickup? PMID- 11937150 TI - Laparoscopic treatment of polycystic ovaries with the holmium:YAG laser. PMID- 11937151 TI - Observer variation study and the potential of power Doppler energy. PMID- 11937155 TI - Pre-analytical errors: everything that happens to the specimen up to the point of testing. PMID- 11937153 TI - Pre-analytical errors: everything that happens to the specimen up to the point of testing. PMID- 11937156 TI - Triterpenoids from Gentiana scabra. AB - Five triterpenoids, (20S)-dammara-13(17),24-dien-3-one, (20R)-dammara-13(17),24 dien-3-one, chirat-16-en-3-one, chirat-17(22)-en-3-one and 17beta,21beta epoxyhopan-3-one, were isolated from the rhizomes and roots of Gentiana scabra together with five known ones, chiratenol, hop-17(21)-en-3-one, hop-17(21)-en 3beta-ol, lupeol and alpha-amyrin. The structures of new compounds were elucidated on the basis of spectroscopic studies. PMID- 11937157 TI - Sesquiterpene constituents in Petasites hybridus. AB - The essential oil of the rhizomes of Petasites hybridus (Asteraceae) was investigated by gas chromatography, mass spectrometry, 1- and 2-dimensional NMR techniques and chemical correlations. Two new sesquiterpene hydrocarbons, petasitene and pethybrene, could be identified. Petasitene is the parent sesquiterpene hydrocarbon to the known norsesquiterpene albene. The absolute configuration of petasitene could be assigned by conversion of natural albene to petasitene by partial synthesis. Pethybrene is a tricyclic sesquiterpene hydrocarbon, which rearranges to the structurally related alpha-isocomene under acidic conditions. Several sesquiterpenes were isolated from the hydrodistillation products of Petasites hybridus and investigated by spectroscopic methods and chemical correlations PMID- 11937158 TI - epi-Cubebanes from Solidago canadensis. AB - GC-MS of the essential oil prepared by hydrodistillation of the green parts of a specimen of Solidago canadensis collected near Katowice, Poland, revealed two new sesquiterpene hydrocarbons. Their EI mass spectra resembled the mass spectrum of beta-ylangene (1) but the retention indices of the new compounds differed markedly from this known compound. After isolation of the new compounds by preparative GC their investigation by one- and two-dimensional NMR techniques resulted in the identification of 6-epi-alpha-cubebene (2) (minor constituent, 1.5%) and 6-epi-beta-cubebene (3) (major constituent, 20.5%). PMID- 11937159 TI - Sesquiterpenoids of Torilis japonica fruit. AB - From the methanolic extract of Torilis japonica D. C. fruit (Umbelliferae), two eudesmane-type sesquiterpenoids were isolated together with five previously described sesquiterpenoids. From the results of spectral analyses, they were characterized as 4(15)-eudesmene-1beta,5alpha-diol and 4alpha,15-epoxyeudesmane 1beta,6alpha-diol, respectively. The absolute stereostructures of these sesquiterpenoids were elucidated by the modified Mosher's method. PMID- 11937160 TI - Sphaeropsidins D and E, two other pimarane diterpenes, produced in vitro by the plant pathogenic fungus Sphaeropsis sapinea f. sp. cupressi. AB - Two pimarane diterpenes structurally related to sphaeropsidins were isolated from the liquid culture of Sphaeropsis sapinea f. sp. cupressi, a plant pathogenic fungus causing a form of canker disease of Italian cypress (Cupressus sempervirens L.). The two metabolites, characterised by spectroscopic methods, were named sphaeropsidins D (0.40 mg l(-1)) and E (0.16 mg l(-1)). The same fungus produced sphaeropsidins A, B and C, sphaeropsidone and episphaeropsidone, which proved to be phytotoxic to cypress, and chlorosphaeropsidone and epichlorosphaeropsidone showing no phytotoxicity. Sphaeropsidin D assayed at 0.1 mg ml(-1) on severed cypress twigs caused leaf browning and necrosis on Cupressus macrocarpa, but no symptoms were observed on C. sempervirens and C. arizonica. Symptoms appeared in a period of time (6 days after toxin-treatment) shorter than that for sphaeropsidin A. Sphaeropsidin E assayed at 0.2 mg ml(-1) did not produce any symptom on the same cypress species tested with sphaeropsidin D. PMID- 11937161 TI - Triterpenoid saponins and acylated prosapogenins from Harpullia austro caledonica. AB - Three new triterpenoid saponins have been isolated from the stem bark of Harpullia austro-caledonica and identified as 24-O-[alpha-L-rhamnopyranosyl-(1 >2)-beta-D-glucopyranosyl]-28-O-[beta-D-glucopyranosyl-(1->2)-beta-D glucopyranosyl]-protoaescigenin, 24-O-[alpha-L-rhamnopyranosyl-(1->2)-beta-D glucopyranosyl]-28-O-[beta-D-glucopyranosyl-(1->2)-beta-D-glucopyranosyl]-16 desoxyprotoaescigenin, 24-O-[alpha-L-rhamnopyranosyl-(1->2)-beta-D glucopyranosyl]-28-O-[beta-D-glucopyranosyl-(1->2)-beta-D-glucopyranosyl]-24-oxo camelliagenin D. The 21,22-di-O-angeloate esters of protoaescigenin and barringtogenol C were isolated in the acid hydrolysate of the saponin extract together with a new prosapogenin identified as 21beta,22alpha-di-O-angeloyl camelliagenin D. The structures were established using one- and two- dimensional NMR and mass spectrometry. PMID- 11937162 TI - Quassinoids from Eurycoma longifolia. AB - Three quassinoids, eurycolactone D (1), eurycolactone E (2) and eurycolactone F (3) were isolated from the roots of Eurycoma longifolia Jack. The structures of 1 3 were elucidated by spectroscopic methods, and that of 3 was further confirmed by X-ray crystallography. The known quassinoids, laurycolactone B (4) and eurycomalactone (5) were also identified. PMID- 11937163 TI - Botryolins A and B, two tetramethylsqualene triethers from the green microalga Botryococcus braunii. AB - Two new triterpenoid polyethers with a tetramethylsqualene carbon skeleton, botryolins A and B, have been isolated from the green microalga Botryococcus braunii. Their structures were determined by means of spectral analyses including 2D NMR. PMID- 11937164 TI - Guaiane- and aristolane-type sesquiterpenoids of Nardostachys chinensis roots. AB - Two guaiane-type compounds, nardoguaianone J and K (1 and 2) and two aristolane type compounds, kanshone F and G (3 and 4), were isolated from Nardostachys chinensis roots. The structures including the absolute configurations were elucidated by spectral means and by comparison of their CD spectra. PMID- 11937165 TI - Two highly oxygenated eudesmanes and 10 lignans from Achillea holosericea. AB - Two new highly oxygenated eudesmanes and 10 known lignans were isolated from the aerial parts of Achillea holosericea. Their structures were elucidated by extensive application of one- and two-dimensional 1H and 13C NMR spectroscopy. PMID- 11937166 TI - Phenolic glycosides from Phagnalon rupestre. AB - Analysis of the butanol-soluble fraction from the methanolic extract of the aerial parts of Phagnalon rupestre (Asteraceae) has led to the isolation of seven phenolic compounds. Three have been identified on the basis of their NMR spectra as new natural compounds: the lignan 7,7'-bis-(4-hydroxy-3,5-dimethoxyphenyl) 8,8'-dihydroxymethyl-tetrahydrofuran-4-O-beta-glucopyranoside (1), the prenylhydroquinone glycoside 1-O-beta-glucopyranosyl-1,4-dihydroxy-2-(3'-hydroxy 3'-methylbutyl) benzene (2) and the acetophenone glycoside 12-O-beta glucopyranosyl-9beta,12-dihydroxytremetone (3). The known flavonoids apigenin-7-O beta-glucoside, luteolin-7-O-beta-glucoside, luteolin-7-O-beta-glucuronide and the acetophenone picein were also isolated. PMID- 11937167 TI - Constituents from the stems of Actinodaphne lancifolia. AB - Two C(16)-lactonic compounds, actinolides A-B (1-2), were isolated from the stems of Actinodaphne lancifolia, together with five known lactones (3-7) and three known lignans (8-10). Their structures were determined spectroscopically, which included 2D NMR spectroscopic analysis. PMID- 11937168 TI - Two benzophenone O-arabinosides and a chromone from Hypericum annulatum. AB - Two benzophenone O-arabinosides, annulatophenonoside (1) and acetylannulatophenonoside (2) were isolated from the methanol extract of the herb of Hypericum annulatum. The structures of the benzophenones were established as 2 O-alpha-L-arabinofuranosyl-3',5',6-trihydroxy-4-methoxybenzophenone (1) and 2-O alpha-L-3"-acetylarabinofuranosyl-3',5',6-trihydroxy-4-methoxybenzophenone (2) based on spectral and chemical evidence. A chromone, 5,7-dihydroxy-3 methylchromone (3) was isolated from the chloroform extract. Although it has been previously synthesized it is encountered in a plant source for the first time. Co occurrence of the two new benzophenone O-arabinosides along with the biogenetically related 1,5,7-trihydroxy-3-methoxyxanthone was not found. PMID- 11937169 TI - Three anthrones from Rubus ulmifolius. AB - From the aerial parts of Rubus ulmifolius Schott three new anthrones, rubanthrone A, B and C, have been isolated. Their structures were established by spectral procedures including 1D and 2D NMR techniques and chemical derivatization. Rubanthrone A showed antimicrobial activity against Staphylococcus aureus at 4.5 mg/ml. PMID- 11937170 TI - Chalcones and other constituents of Dorstenia prorepens and Dorstenia zenkeri. AB - The twigs of Dorstenia prorepens furnished the digeranylated chalcone, 5,3'-(3,7 dimethyl-2,6-octadienyl)-3,4, 2',4'-tetrahydroxychalcone while Dorstenia zenkeri yielded the 3',4'-(3-hydroxy-2,2-dimethyldihydropyrano)-4,2'-dihydroxychalcone and a bichalcone. 4-Hydroxylonchocarpin was found in both plants. D. prorepens also yielded the known compounds: psoralen, bergapten, beta-sitosterol and its D glucopyranosyl derivative. D. zenkeri yielded p-hydroxybenzaldehyde, dorsmanin A, 4,2',4'-trihydroxychalcone and 4,2',4'-trihydroxy-3'-prenylchalcone. Structures of the new compounds were established by UV, IR, MS and 2-D NMR analysis. PMID- 11937171 TI - Klaivanolide, an antiprotozoal lactone from Uvaria klaineana. AB - Bioguided-fractionation of a CH(2)Cl(2) extract of the stems of Uvaria klaineana (Annonaceae) led to isolation of klaivanolide, a novel bisunsaturated 7-membered lactone (5-acetoxy-7-benzoyloxymethyl-7H-oxepin-2-one), together with benzyl benzoate. Klaivanolide showed potent in vitro antileishmanial activity against both sensitive and amphotericin B-resistant promastigote forms of Leishmania donovani with IC(50) values of 1.75 and 3.12 microM, respectively. The compound also showed in vitro trypanocidal activity against trypomastigote forms of Trypanosoma brucei brucei GVR 35. Its structure was established by 1D and 2D NMR and other spectroscopic techniques. PMID- 11937172 TI - A chlorinated monoterpene ketone, acylated beta-sitosterol glycosides and a flavanone glycoside from Mentha longifolia (Lamiaceae). AB - Mentha longifolia (Lamiaceae), an aromatic herb yielded a new halogenated chloro derivative of menthone (longifone), two new derivatives of beta-sitosterol glycoside (longiside-A and -B) and a new flavanone-glycoside (longitin). The beta sitosterol and flavanone glycosides were purified as their acetate derivatives. Structures of all the isolated constituents were elucidated with the aid of HMBC techniques. However, the structure of longifone was also determined through X-ray crystallography. PMID- 11937173 TI - Amides from Piper brachystachyum and Piper retrofractum. AB - Three unsaturated amides, designated brachystamides-C, D and E have been characterised from Piper brachystachyum Wall. Brachystamide-C, shown to be N isobutyl-15-(3',4'-methylenedioxyphenyl)-2E,4E,13E-pentadecatrienamide, was unusual in having a non-conjugated double bond. Piper retrofractum Vahl. yielded retrofractamide-D, which has been fully characterised. PMID- 11937174 TI - Feeding the world's children. PMID- 11937175 TI - Angiotensin blockade for hypertension: a promise fulfilled. PMID- 11937176 TI - Vertical HIV transmission in South Africa: translating research into policy and practice. PMID- 11937177 TI - The health (and wealth) of nations. PMID- 11937178 TI - Cardiovascular morbidity and mortality in the Losartan Intervention For Endpoint reduction in hypertension study (LIFE): a randomised trial against atenolol. AB - BACKGROUND: Blood pressure reduction achieved with beta-blockers and diuretics is the best recorded intervention to date for prevention of cardiovascular morbidity and death in patients with hypertension. Left ventricular hypertrophy (LVH) is a strong independent indicator of risk of cardiovascular morbidity and death. We aimed to establish whether selective blocking of angiotensin II improves LVH beyond reducing blood pressure and, consequently, reduces cardiovascular morbidity and death. METHODS: We did a double-masked, randomised, parallel-group trial in 9193 participants aged 55-80 years with essential hypertension (sitting blood pressure 160-200/95-115 mm Hg) and LVH ascertained by electrocardiography (ECG). We assigned participants once daily losartan-based or atenolol-based antihypertensive treatment for at least 4 years and until 1040 patients had a primary cardiovascular event (death, myocardial infarction, or stroke). We used Cox regression analysis to compare regimens. FINDINGS: Blood pressure fell by 30.2/16.6 (SD 18.5/10.1) and 29.1/16.8 mm Hg (19.2/10.1) in the losartan and atenolol groups, respectively. The primary composite endpoint occurred in 508 losartan (23.8 per 1000 patient-years) and 588 atenolol patients (27.9 per 1000 patient-years; relative risk 0.87, 95% CI 0.77-0.98, p=0.021). 204 losartan and 234 atenolol patients died from cardiovascular disease (0.89, 0.73-1.07, p=0.206); 232 and 309, respectively, had fatal or non-fatal stroke (0.75, 0.63 0.89, p=0.001); and myocardial infarction (non-fatal and fatal) occurred in 198 and 188, respectively (1.07, 0.88-1.31, p=0.491). New-onset diabetes was less frequent with losartan. Interpretation Losartan prevents more cardiovascular morbidity and death than atenolol for a similar reduction in blood pressure and is better tolerated. Losartan seems to confer benefits beyond reduction in blood pressure. PMID- 11937179 TI - Cardiovascular morbidity and mortality in patients with diabetes in the Losartan Intervention For Endpoint reduction in hypertension study (LIFE): a randomised trial against atenolol. AB - BACKGROUND: The most suitable antihypertensive drug to reduce the risk of cardiovascular disease in patients with hypertension and diabetes is unclear. In prespecified analyses, we compared the effects of losartan and atenolol on cardiovascular morbidity and mortality in diabetic patients. METHODS: As part of the LIFE study, in a double-masked, randomised, parallel-group trial, we assigned a group of 1195 patients with diabetes, hypertension, and signs of left ventricular hypertrophy (LVH) on electrocardiograms losartan-based or atenolol based treatment. Mean age of patients was 67 years (SD 7) and mean blood pressure 177/96 mm Hg (14/10) after placebo run-in. We followed up patients for at least 4 years (mean 4.7 years [1.1]). We used Cox regression analysis with baseline Framingham risk score and electrocardiogram-LVH as covariates to compare the effects of the drugs on the primary composite endpoint of cardiovascular morbidity and mortality (cardiovascular death, stroke, or myocardial infarction). FINDINGS: Mean blood pressure fell to 146/79 mm Hg (17/11) in losartan patients and 148/79 mm Hg (19/11) in atenolol patients. The primary endpoint occurred in 103 patients assigned losartan (n=586) and 139 assigned atenolol (n=609); relative risk 0.76 (95% CI 0.58-.98), p=0.031. 38 and 61 patients in the losartan and atenolol groups, respectively, died from cardiovascular disease; 0.63 (0.42 0.95), p=0.028. Mortality from all causes was 63 and 104 in losartan and atenolol groups, respectively; 0.61 (0.45-0.84), p=0.002. INTERPRETATION: Losartan was more effective than atenolol in reducing cardiovascular morbidity and mortality as well as mortality from all causes in patients with hypertension, diabetes, and LVH. Losartan seems to have benefits beyond blood pressure reduction. PMID- 11937180 TI - Chemotherapy in adult high-grade glioma: a systematic review and meta-analysis of individual patient data from 12 randomised trials. AB - BACKGROUND: Trials on the effect of systemic chemotherapy on survival and recurrence in adults with high-grade glioma have had inconclusive results. We undertook a systematic review and meta-analysis to assess the effects of such treatment on survival and recurrence. METHODS: We did a systematic review and meta-analysis using updated data on individual patients from all available randomised trials that compared radiotherapy alone with radiotherapy plus chemotherapy. Data for 3004 patients from 12 randomised controlled trials were included (11 published and one unpublished). FINDINGS: Overall, the results showed significant prolongation of survival associated with chemotherapy, with a hazard ratio of 0.85 (95% CI 0.78-0.91, p<0.0001) or a 15% relative decrease in the risk of death. This effect is equivalent to an absolute increase in 1-year survival of 6% (95% CI 3-9) from 40% to 46% and a 2-month increase in median survival time (1-3). There was no evidence that the effect of chemotherapy differed in any group of patients defined by age, sex, histology, performance status, or extent of resection. INTERPRETATION: This small but clear improvement in survival from chemotherapy encourages further study of drug treatment of these tumours. PMID- 11937181 TI - Mortality associated with Down's syndrome in the USA from 1983 to 1997: a population-based study. AB - BACKGROUND: Down's syndrome is the most frequently identified cause of mental retardation, but information about mortality and comorbidity in people with Down's syndrome is limited. METHODS: We used data from US death certificates from 1983 to 1997 to calculate median age at death and standardised mortality odds ratios (SMORs) for common medical disorders in people with Down's syndrome. FINDINGS: Of 17897 people reported to have Down's syndrome, median age at death increased from 25 years in 1983 to 49 years in 1997, an average increase of 1.7 years per year studied (p<0.0001). Median age at death was significantly lower in black people and people of other races than in white people with Down's syndrome. As expected, death certificates with a diagnosis of Down's syndrome were more likely to list congenital heart defects (SMOR 29.1, 95% CI 27.8-30.4), dementia (21.2, 19.6-22.7), hypothyroidism (20.3, 18.5-22.3), or leukaemia (1.6, 1.4-1.8) than were those that did not report Down's syndrome. By contrast, malignant neoplasms other than leukaemia were listed on death certificates of people with Down's syndrome less than one-tenth as often as expected (0.07, 0.06-0.08). A strikingly low SMOR for malignancy was associated with Down's syndrome at all ages, in both sexes, and for all common tumour types except leukaemia and testicular cancer. INTERPRETATION: Identification of factors responsible for the racial differences recorded could facilitate further improvement in survival of people with Down's syndrome. Reduced exposure to environmental factors that contribute to cancer risk, tumour-suppressor genes on chromosome 21, or a slower rate of replication or higher likelihood of apoptosis in Down's syndrome cells, could be possible reasons for paucity of cancer in people with Down's syndrome. PMID- 11937182 TI - Clinical picture: carcinoma erysipeloides of the neck. PMID- 11937183 TI - Association between altered expression of adipogenic factor SREBP1 in lipoatrophic adipose tissue from HIV-1-infected patients and abnormal adipocyte differentiation and insulin resistance. AB - BACKGROUND: Lipodystrophy is a major side-effect of antiretroviral therapy but its pathophysiology remains elusive. In-vitro studies show that HIV-1-protease inhibitors affect adipocyte differentiation at an early step involving sterol regulatory-element-binding-protein-1 (SREBP1), but in-vivo studies are lacking. METHODS: We compared fat morphology and mRNA and protein expression of major adipocyte differentiation markers and cytokines in subcutaneous abdominal adipose tissue from 26 HIV-1-infected patients who developed peripheral lipoatrophy while on protease inhibitors and from 18 HIV-1-seronegative healthy controls. FINDINGS: Patients' fat contained a higher proportion of small adipocytes than control fat, together with lower mRNA concentrations of the adipogenic differentiation factors CCAAT-enhancer binding protein (C/EBP) beta and alpha, peroxisome proliferator activated receptor (PPAR) gamma, and the 1c isoform of SREBP1, with a median decrease of 93% in the latter. The SREBP1 protein concentration was increased 2.6 fold, whereas the PPARgamma protein concentration was decreased by 70%. The expression of adipocyte-specific markers, including leptin, was lower in fat from patients than in fat from controls, whereas expression of tumour necrosis factor (TNF) alpha was higher and correlated negatively with the expression of SREBP1c and downstream adipogenic factors. SREBP1c mRNA concentrations correlated negatively, and TNFalpha mRNA concentrations positively, with glycaemia and insulin resistance, but did not correlate with lipid variables. INTERPRETATION: The altered differentiation status of peripheral adipocytes in HIV-1-infected patients with antiretroviral-induced lipoatrophy is associated with greatly reduced SREBP1c expression. Since the differentiation factor SREBP1 is rapidly targeted by protease inhibitors in vitro, our results suggest that SREBP1c could be an important mediator of peripheral lipoatrophy in this setting, leading to metabolic alterations such as insulin resistance. PMID- 11937184 TI - A child with oesophageal strictures. PMID- 11937185 TI - Polymorphism of the thymidylate synthase gene and outcome of acute lymphoblastic leukaemia. AB - Interindividual variability in response to methotrexate could be caused by variable concentrations of thymidylate synthase. We investigated the possible association between a tandem-repeat polymorphism in the thymidylate synthase promoter, of which a triple repeat is associated with increased expression of thymidylate synthase, and outcome of acute lymphoblastic leukaemia in 205 children treated with methotrexate. We obtained DNA samples from buccal epithelial cells, peripheral blood, or bone marrow in remission, and analysed them for the polymorphism by PCR amplification. Individuals who were homozygous for the triple repeat had a poorer outlook than those with other genotypes (odds ratio 4.1, 95% CI 1.9-9.0, p=0.001). Genotyping of thymidylate synthase might make it possible to individualize treatment for patients with acute lymphoblastic leukaemia. PMID- 11937186 TI - Antibodies to varicella-zoster virus in blood donors with genetic variance in CC chemokine receptor 5. AB - Carriers of a 32 bp deletion (Delta32) allele of the CC chemokine receptor 5 (CCR5) gene are reported to be more likely to lack antibodies to varicella-zoster virus than CCR5 wild-type individuals. To find out whether CCR5-Delta32 is associated with the seroprevalence of varicella-zoster virus infection, we tested blood donors with different CCR5-Delta32 genotypes for varicella-zoster virus IgG. Antibody to varicella-zoster virus was present in 209 (99.5%) of 210 CCR5 Delta32 carriers and exactly the same proportion of CCR5 wild-type individuals (209 of 210). We have therefore found no evidence that the CCR5-Delta32 allele is associated with decreased seroprevalence of varicella-zoster virus infection. PMID- 11937187 TI - Association between genetic variation in the gene for insulin-like growth factor I and low birthweight. AB - Low birthweight is associated with later risk of type 2 diabetes and related disorders. We aimed to show that a polymorphism in the gene for insulin-like growth factor-I, which has proved to raise risk of type 2 diabetes and myocardial infarction, is associated with low birthweight. We recorded birthweight and obtained DNA for 463 adults. Individuals who did not have the wild-type allele of the polymorphism had a 215 g lower birthweight than those homozygous for this allele (95% CI -411 to -10). Our data lend support to the hypothesis that genetic variation affecting fetal growth could account for the association between low birthweight and susceptibility to diabetes and cardiovascular disease in later life. PMID- 11937188 TI - Fatal non-cardiogenic pulmonary oedema after intravenous non-ionic radiographic contrast. AB - Life-threatening reactions to radiographic contrast media are rare. We describe a case of fatal non-cardiogenic pulmonary oedema, after use of iopamidol, a widely used, low osmolar, non-ionic, radiographic contrast medium. A woman with a history of bladder tumour who had haematuria was referred for an intravenous urogram. After intravenous injection of 100 mL iopamidol, the woman became short of breath, and a chest radiograph showed severe pulmonary oedema. She was treated but subsequently died. The pathogenesis of contrast-induced, non-cardiogenic pulmonary oedema is unclear, and treatment differs from that for cardiogenic pulmonary oedema. PMID- 11937189 TI - German doctors' links with drug firm investigated. PMID- 11937190 TI - Researchers describe autoimmune mechanism for rheumatoid arthritis. PMID- 11937193 TI - Orally available cidofovir derivative active against smallpox. PMID- 11937194 TI - Stimulating research in the most neglected diseases. PMID- 11937195 TI - London--The NHS-the problem is capacity, not funding. PMID- 11937196 TI - Scurvy outbreak in Afghanistan prompts food aid concerns. PMID- 11937198 TI - WHO's tobacco control chairman urges faster progress at FCTC talks. PMID- 11937199 TI - Health-rights organisation wants Uganda's intellectual property law delayed. PMID- 11937200 TI - An Olympic gold standard in sports medicine. PMID- 11937201 TI - Stem cells and neuropoiesis in the adult human brain. AB - Stem cells in adult tissues have attracted a great deal of interest. These cells are self-renewing and can give rise to diverse progeny. An extraordinary finding was the presence of stem cells in the mature human brain. This tissue was previously believed incapable of generating new neurons, but neuropoiesis is now an established phenomenon in the adult brains of mammals, including human beings. This persistent neurogenesis has potential therapeutic applications for various neurological disorders as a source for tissue engraftment and as self-repair by a person's own indigenous population of pluripotent cells or biogenic by-products of their proliferation and differentiation. PMID- 11937202 TI - Ethical analysis in public health. AB - Public-health regularly encounters serious ethical dilemmas, such as rationing scarce resources, influencing individuals to change their behaviour, and limiting freedom to diminish disease transmission. Yet unlike medical ethics, there is no agreed-upon framework for analysing these difficulties. We offer such a framework. It distinguishes three philosophical views, often invoked in public health discourse: positions based on outcomes (utilitarianism), positions focused on rights and opportunities (liberalism), and views that emphasise character and virtue (communitarianism). We explore critical variations within each approach, and identify practical problems that arise in addressing the ethical dimensions of health policy. We conclude by examining challenges posed by the feminist argument of ethics-of-care and by postmodern views about the nature of ethics. Health professionals need enhanced skills in applied philosophy to improve the coherence, transparency, and quality of public deliberations over ethical issues inherent in health policy. PMID- 11937203 TI - Pott's puffy tumour. PMID- 11937204 TI - Uses of error: ambylopia and arrogance. PMID- 11937205 TI - Primary interferon resistance and treatment response in chronic hepatitis C infection. PMID- 11937207 TI - Cost-effectiveness of diagnostic tests. PMID- 11937208 TI - Ecological studies, rotavirus vaccination, and intussusception. PMID- 11937210 TI - Ecological studies, rotavirus vaccination, and intussusception. PMID- 11937211 TI - Parasite saliva as a source of antiallergic agents. PMID- 11937212 TI - Laboratory practice and studies of bovine spongiform encephalopathy. PMID- 11937213 TI - Preoperative and postoperative radiotherapy and survival in colorectal cancer. PMID- 11937214 TI - Preoperative and postoperative radiotherapy and survival in colorectal cancer. PMID- 11937215 TI - Calcification in chronic renal failure. PMID- 11937217 TI - Peripheral arterial disease. PMID- 11937219 TI - Reconstruction of healthcare in Afghanistan. PMID- 11937220 TI - Reconstruction of health care in Afghanistan. PMID- 11937221 TI - Reconstruction of healthcare in Afghanistan. PMID- 11937222 TI - An old meaning of the word apoptosis. PMID- 11937230 TI - Hearing is believing. PMID- 11937232 TI - Comparison of the disease incidences of dairy cows kept in cold and warm loose housing systems. AB - Finland's cold loose-housing systems for dairy cows were compared with the more traditional warm loose-housing systems regarding the incidences of ketosis, mastitis, metritis, parturient paresis and ovarian disorders. Approximately 5000 calvings on 210 farms during the years 1996 and 1997 were modelled, using multilevel Poisson regression and multilevel logistic-regression in a retrospective observational cohort study. Cows in a cold loose-housing system were at lower odds for developing late mastitis (15-305 days in milk), and metritis (Friesian breed); of the same odds for ketosis and early mastitis (0-14 days in milk); but at higher odds for developing parturient paresis and metritis (Ayrshire breed). The estimated odds ratio for ovarian disorders depended on the definition for exposure. Although one of the differences was statistically significant and many of them of veterinary interest, none of them appear to be substantial for the economy of a median-sized dairy farm in Finland. PMID- 11937233 TI - Factors associated with Neospora caninum serostatus in cattle of 20 specialised Costa Rican dairy herds. AB - Twenty-five specialised Costa Rican dairy farms (located in the Poas area) were used to determine neosporosis seroprevalence and the association of seropositivity with environmental and management factors. The farms involved were selected intentionally and all of them use VAMPP 5.1 (Veterinary Automated Management and Production Control Programme) as management-information system. Holstein-Friesian, Jersey and crosses between them were the most-frequent breeds in these herds. The number of females per farm varied from 41 to 296. Our cross sectional study had two phases. In the first phase, we determined the presence or absence of seropositivity at herd level. For the second phase, all females in 20 seropositive farms were bled. Serum samples were tested for antibodies to Neospora caninum using an indirect enzyme-linked immunosorbent assay (ELISA). A questionnaire with factors mentioned in the literature was administered to the farmers. Logistic regression (LR with herd as random effect) was used to assess the relationships of the serostatus at the individual level with characteristics of the cows and environmental factors. In the first phase all herds had >20% seropositive females; therefore, all herds were eligible for the second phase. In the second phase, the overall prevalence was 39.7% (1191/3002), and within-herd prevalences were between 25.0 and 70.5%. Age 3-6 years, parity < or =2 of the dam of the cow, Jersey breed and lack of purposive sampling to diagnose abortive infectious disease were associated with positive serostatus; other management and environmental factors did not show significant associations. The lack of association between management and environmental factors with serostatus might be because all farms were exposed to a considerable number of potential factors. That all herds of this study were seropositive for neosporosis and the within herd prevalence was considerable raises questions about how far the infection is spread in other dairy areas of Costa Rica. PMID- 11937234 TI - Participatory analysis of seasonal incidences of diseases of cattle, disease vectors and rainfall in southern Sudan. AB - During an investigation into a chronic wasting disease in southern Sudanese cattle, a participatory appraisal method called a 'seasonal calendar' was used to understand local perceptions of seasonal variations in cattle diseases, disease vectors, intermediate hosts and rainfall. Repetition of a standardized seasonal calendar with Dinka informants demonstrated good reproducibility of the method. Comparison of rainfall data produced by seasonal calendars and objective measures of rainfall demonstrated good validity of the seasonal calendar method. Subjective assessment of seasonal calendar scoring patterns by veterinarians indicated that herders' perceptions of seasonal populations of biting flies, ticks and snails were similar to modern veterinary knowledge. The uses of seasonal calendars in veterinary epidemiology are discussed. PMID- 11937235 TI - Risk factors for stillbirths in two swine farms in the south of Brazil. AB - We evaluated stillbirth risk factors in two commercial swine farms of the Rio Grande do Sul State (south of Brazil). The study was conducted during 1 month in Farm A and during 2 months in Farm B, both during 1999. Data for all farrowings that occurred during the study period were recorded (101 for Farm A and 373 for Farm B), without interference in the farm management. In Farm A, 39% of all litters born during the period of interest had stillborn piglets and the stillborn risk for piglets was 12%. In Farm B, 25% of all litters had stillborn piglets whereas the stillborn risk was 2%. Variables considered as potential risk factors for stillbirths were: parity (1, 2-3, 4+); breed (purebred or crossbred); sow body-condition (normal or fat); use of oxytocin during parturition (yes or no); obstetric intervention through vaginal palpation (yes or no); farrowing duration (<4 or > or =4h); mummified fetuses (yes or no); total litter size (<12 or > or =12 piglets); and litter birth weight (<11 or > or =11kg). All stillborn piglets had their classification validated by necropsy. In multivariable logistic regressions, the cases were the litters having at least one stillborn piglet. In Farm A, litters having at least 12 pigs and in which oxytocin was used during the parturition had 20.8-times-higher odds of stillborn occurrence. In Farm B, litters from sows having parity > or =4 had 2.2-times-higher odds of stillborn occurrence than litters from parity 2 to 3 females, litters having > or =12 pigs had 2.0-times-higher odds of a stillborn piglet than smaller litters and farrowings in which vaginal palpation was performed had 8.0-times-higher odds. Farrowing room management to minimize stillborn risk should target higher-parity females, large litters and optimization of practices of obstetric interventions. PMID- 11937236 TI - Risk factors for gastric ulcers in Australian pigs at slaughter. AB - The stomachs of pigs (n=15,741) originating from 136 herds from the Australian states of Queensland, Western Australia, Victoria and New South Wales were examined at slaughter for the presence of oesophago-gastric ulcers (OGUs). Stomachs were categorised as being normal, hyperkeratotic, eroded, ulcerated, or having strictures. A questionnaire was distributed to piggery owners to identify factors associated with an above-average herd prevalence of OGU. Thirty percent of all pigs examined had OGU (median within-herd prevalence of 17%). The median within-herd prevalence in Victoria (53%) was significantly higher than in Western Australia (30%) or Queensland (7%). The prevalence of OGU in culled breeding animals was significantly higher than in porkers or baconers from the same herds. There was no difference between the prevalence of OGU in male and female pigs sampled from the same Western Australian herds. The relationship between OGU and herd and pig risk factors was assessed by random effects logistic-regression analysis. Herds with a high prevalence of OGU were more likely to feed ad libitum (OR=13.7), use automated feeding systems (OR=7.8), feed a pelleted ration (OR=384) and get water from a dam rather than from a bore or river (OR=3.8). Furthermore, for every change in the ration formulation for finisher pigs, the risk of OGU increased 1.5 times. PMID- 11937237 TI - Flock-level prevalence of Eimeria species among broiler chicks in northern Jordan. AB - Six chicks (3-6 weeks of age) were taken randomly from each of 200 broiler farms in northern Jordan, these chicks were submitted for post-mortem and parasitological examinations. Seven Eimeria spp. were identified: E. acervulina, E. brunetti, E. maxima, E. necatrix, E. mivati, E. mitis, and E. tenella. Half (50%) of the farms surveyed had all six chicks infected, 23% of the farms were free of the infection. E. tenella was the most prevalent species (39%) followed by E. necatrix (12%), E. brunitti (12%), and E. maxima (10%). Prevalences did not vary by flock size. Also, neither the use of coccidiostat nor previous coccidiosis clinical outbreaks was associated with the prevalence of coccidiosis. PMID- 11937238 TI - Herd-level seroprevalence of swine-influenza virus in Korea. AB - A total of 911 serum samples from 130 herds (an average of nine serum samples per herd) in Korea were examined for antibody to swine H1N1-influenza virus using enzyme-linked immunosorbent assay (ELISA). The list of farms was obtained from the Korean Swine Association, and herds were included from all five of the country's states. Farms were selected using a random-numbers table for swine within farms and for farms. All serum samples were collected from 22- to 24-week old finishing pigs between September 2000 and March 2001. By ELISA, 93 out of 130 sampled herds (71.5%) were positive against swine H1N1-influenza virus. Our data suggested that seropositive herds for swine H1N1-influenza virus are distributed diffusely throughout the Republic of Korea. PMID- 11937239 TI - Intensity modulated versus non-intensity modulated radiotherapy in the treatment of the left breast and upper internal mammary lymph node chain: a comparative planning study. AB - BACKGROUND AND PURPOSE: To compare and evaluate intensity modulated (IMRT) and non-intensity modulated radiotherapy techniques in the treatment of the left breast and upper internal mammary lymph node chain. MATERIALS AND METHODS: The breast, upper internal mammary chain (IMC), heart and lungs were delineated on a computed tomography (CT)-scan for 12 patients. Three different treatment plans were created: (1) tangential photon fields with oblique IMC electron-photon fields with manually optimized beam weights and wedges, (2) wide split tangential photon fields with a heart block and computer optimized wedge angles, and (3) IMRT tangential photon fields. For the IMRT technique, an inverse planning program (KonRad) generated the intensity profiles and a clinical three dimensional treatment planning system (U-MPlan) optimized the segment weights. U MPlan calculated the dose distribution for all three techniques. The normal tissue complication probabilities (NTCPs) for the organs at risk (ORs) were calculated for comparison. RESULTS: The average root mean square deviation of the differential dose-volume histogram of the breast planning target volume was 4.6, 3.9 and 3.5% and the average mean dose to the IMC was 97.2, 108.0 and 99.6% for the oblique electron, wide split tangent and IMRT techniques, respectively. The average NTCP for the ORs (i.e. heart and lungs) were comparable between the oblique electron and IMRT techniques (or=2%) for the ORs. CONCLUSIONS: The lowest NTCP values were found with the oblique electron and the IMRT techniques. The IMRT technique had the best breast and IMC target coverage. PMID- 11937240 TI - Critical appraisal of treatment techniques based on conventional photon beams, intensity modulated photon beams and proton beams for therapy of intact breast. AB - PURPOSE: To analyse different treatment techniques with conventional photon beams, intensity modulated photon beams, and proton beams for intact breast irradiation for patients in whom conventional irradiation would cause potentially dangerous lung irradiation. MATERIALS AND METHODS: Five breast cancer patients with highly concave breast tissue volume around the lung were considered at planning level in order to assess the suitability of different irradiation techniques. Three-dimensional dose distributions for conventional two-field tangential photon treatment, two-field intensity modulated radiotherapy (IMRT), three-field non-IMRT, three-field IMRT, and single-field proton treatment were investigated, aiming at assessing the possibility to reduce lung irradiation below risk levels. Analysis of dose-volume histograms and related physical and biological parameters (significant minimum, maximum and mean doses, conformity indexes and equivalent uniform dose (EUD)) for planned target volume (PTV) and lung was carried out. Dose plans were compared with the conventional two-field tangential photon technique. RESULTS: PTV coverage was comparable for non-IMRT and IMRT techniques (EUD from 47.1 to 49.4 Gy), and improved with single-field proton treatment (EUD=49.8 Gy). Lung irradiation was reduced, in terms of mean dose, with three-field (9.5 Gy) and proton technique (3.5 Gy), with respect to the conventional two-field treatment (12.9 Gy); also a reduction of the lung volume irradiated at high doses was observed. Better results could be achieved with protons. In addition, cardiac irradiation was also reduced with those techniques. CONCLUSIONS: Geometrically difficult breast cancer patients could be irradiated with a three-field non-IMRT technique thus reducing the dose to the lung which is proposed as standard for this category of patients. Intensity modulated techniques were only marginally more successful than the corresponding non-IMRT treatments, while protons offer excellent results. PMID- 11937241 TI - The use of a compensator library to reduce dose inhomogeneity in tangential radiotherapy of the breast. AB - BACKGROUND AND PURPOSE: The dose variation throughout the volume of the breast from tangential fields can exceed 20% for large breasts. This is postulated to result in poor cosmesis [Radiother Oncol 16 (1989) 253], particularly at the inframammary fold, where the dose is highest. Compensators may be used to reduce this variation, but at the cost of the time to manufacture each unique compensator for the individual patients. This paper outlines the implementation and routine use of a library of reusable compensators. MATERIALS AND METHODS: For the period of December 1999 to May 2001, 94 patients attending for breast radiotherapy received treatment using breast compensators calculated from multiple outlines measured using the Osiris system. The compensators manufactured for the early patients were added to a library for possible reuse by later patients. Of the 94 patients, 28 patients' compensators formed the library and 66 subsequent patients have been treated using compensators derived from the library. Selection of the most appropriate library compensator was determined from the analysis of the distribution of the calculated dose-volume histogram for the whole breast, excluding lung, penumbra and build-up regions. Once the library was complete, approximately 50% of all subsequent breast patients were treated with compensators (46% from the library and 4% with individual compensators). This represented a usage rate of 92% for the library compensators for those patients requiring compensation. RESULTS: In all cases the compensators reduced the variation in the dose distribution. For example, the group treated with a library compensator demonstrated a mean reduction from 29 to 9% for the volume of breast tissue receiving more than 5% greater than the reference dose. If the same patients had been treated using their own individual compensators, the corresponding value would have been 7%. There is a small systematic, but negligible, difference in the two populations of dose variation for individual versus library compensators, but this difference (P=0.20) did not reach the level of statistical significance of P=0.05). CONCLUSION: The method of creation and selection of library compensators has proved to be simple and reliable in practice. Every patient receiving radiotherapy for breast cancer is currently investigated under full software control to ascertain whether the use of a library compensator would be advantageous. PMID- 11937242 TI - Lateralisation and depth of the internal mammary chain determined by scintigraphy and by ultrasonography: a comparative study in 124 primary breast cancer patients. AB - In 124 primary breast cancer patients the maximum values of the lateralisation of the internal mammary chain (IMC) and their corresponding depths could be determined by scintigraphy and ultrasonography in 77% and 85% of the cases, respectively. With respect to the lateralisation it appeared that these values were uncorrelated between the two methods. With respect to depth correlation was found (correlation coefficient 0.34; P=0.001). We conclude that substantial differences are noted. Irradiation of the IMC based on computed tomography treatment planning might be a better alternative. PMID- 11937243 TI - Reduction of cardiac and lung complication probabilities after breast irradiation using conformal radiotherapy with or without intensity modulation. AB - PURPOSE: The main purpose of this work is to reduce the cardiac and lung dose by applying conformal tangential beam irradiation of the intact left breast with and without intensity modulation, instead of rectangular tangential treatment fields. The extension of the applicability of the maximum heart distance (MHD) to conformal tangential fields as a simple patient selection criterion, identifying patients for which rectangular and conformal tangential fields without intensity modulation will result in unacceptable normal tissue complication probability (NTCP) values for late cardiac mortality (e.g. >2%), was also investigated. MATERIALS AND METHODS: Three-dimensional treatment planning was performed for 17 left-sided breast cancer patients. Three different tangential beam techniques were compared: (1) optimized wedges without blocks, (2) optimized wedges with conformal blocks and (3) intensity modulation. Plans were evaluated using dose volume histograms (DVHs) for the planning target volume (PTV), the heart and the lungs. NTCPs for radiation pneumonitis and late cardiac mortality were calculated using the DVH data. The MHD was measured for all rectangular (MHD(rectangular)) and conformal (MHD(conformal)) treatment plans. RESULTS: For all patients, on average, part of the PTV receiving a dose between 95 and 107% of the prescribed dose of 50Gy in 25 fractions of 2Gy was 90.8% (standard deviation (SD): 5.0%), 92.8% (SD: 3.5%) and 92.8% (SD: 3.6%) for the intensity modulation radiation therapy (IMRT), conformal and rectangular field treatment techniques, respectively. The NTCP for radiation pneumonitis was 0.3% (SD: 0.1%), 0.4% (SD: 0.4%) and 0.5% (SD: 0.6%) for the IMRT, conformal and rectangular field techniques, respectively. The NTCP for late cardiac mortality was 5.9% (SD: 2.2%) for the rectangular field technique. This value was reduced to 4.0% (SD: 2.3%) with the conformal technique. A further reduction to 2.0% (SD: 1.1%) could be accomplished with the IMRT technique. The NTCP for late cardiac mortality could be described as a second order polynomial function of the MHD. This function could be described with a high accuracy and was independent of the technique for which the MHD was determined (r(2)=0.88). In order to achieve a NTCP value for late cardiac mortality below 1, 2 or 3%, the MHD should be equal to or smaller than 11, 17 or 23 mm, respectively. If such a maximum complication probability cannot be accomplished, a treatment using the IMRT technique should be considered. CONCLUSIONS: The use of conformal tangential fields decreases the NTCP for late cardiac toxicity on average by 30% compared to using rectangular fields, while the tangential IMRT technique can further reduce this value by an additional 50%. The MHD can be used to estimate the NTCP for late cardiac mortality if rectangular or conformal tangential treatment fields are used. PMID- 11937244 TI - Cardiac and pulmonary doses and complication probabilities in standard and conformal tangential irradiation in conservative management of breast cancer. AB - BACKGROUND AND PURPOSE: The clinical benefit of irradiating the intact breast after lumpectomy must be weighted against the risk of severe toxicity. We present a study on cardiac and pulmonary dose-volume data and the related complication probabilities of tangential breast irradiation having the following objectives: (1) to quantify the sparing of the organs at risk (ORs), the heart and the lung, achieved by three-dimensional (3-D) conformal tangential irradiation (CTI) as compared to standard tangential irradiation (STI); (2) to elucidate the uncertainty in radiation tolerance data; and (3) to analyse the relation between the amount of OR irradiated and the resulting morbidity risk. MATERIAL AND METHODS: Computed tomography (CT)-based 3-D treatment plans of 26 patients prescribed to CTI of the intact breast were applied. Contour-based STI has been our routine treatment, and was reconstructed for all patients. Dose-volume data and normal tissue complication probability (NTCP) predictions from the probit and relative seriality models with several cardiac and pulmonary tolerance parameterizations were analysed and compared. RESULTS AND CONCLUSIONS: A significant amount of normal tissues can be spared from radiation by using CT based CTI, resulting in a 50% reduction of the average excess cardiac mortality risk in the left-sided cases. The risks for pericarditis and pneumonitis were too low to reveal any clinically significant difference between the treatments. For the STI set-up, a regression analysis showed that the excess cardiac mortality risk increased when larger parts of the heart were inside the fields. However, the different excess cardiac mortality and pneumonitis tolerance parameters resulted in statistically significant different NTCPs, which precluded the ability to accurately predict absolute NTCPs after tangential breast irradiation. Despite this uncertainty the different series of cardiac and pulmonary risk predictions were in relatively good agreement when small volumes of the ORs were irradiated. From the present data and without consideration of patient or organ motion, it therefore appears that tangential breast irradiation with less than 1 cm of the heart and 2-2.5 cm of the lung included inside the treatment fields will cause at most 1 per thousand risk for cardiac mortality and pulmonary morbidity. CT-based CTI should be considered, in particular for the left-sided cases, if these requirements cannot be met. PMID- 11937245 TI - Acute toxicity of adjuvant doxorubicin and cyclophosphamide for early breast cancer -- a retrospective review of Chinese patients and comparison with an historic Western series. AB - A cohort of 85 Chinese breast cancer patients who received adjuvant chemotherapy with doxorubicin and cyclophosphamide was found to have a significantly higher incidence of grade 3 (n=44, 52%) and grade 4 (n=21, 25%) neutropenia when compared with an historic Western cohort. Also noted was a higher incidence of hepatotoxicity (n=8, 9%). When compared to Caucasian patients, the higher myelotoxicity in our patients may be related to ethnic variation in susceptibility to chemotherapy-related toxicity, lower body mass index with higher percentage of body fat composition, and the popular practice of concurrent alternative medicine during chemotherapy. The higher incidence of hepatoxicity was possibly associated with endemic chronic hepatitis B infection in this geographical area. PMID- 11937246 TI - Up-front centralized data review and individualized treatment proposals in a multicenter pediatric Hodgkin's disease trial with 71 participating hospitals: the experience of the German-Austrian pediatric multicenter trial DAL-HD-90. AB - BACKGROUND AND PURPOSE: A systematic procedure for up-front centralized data review and the set-up of individualized treatment proposals was integrated prospectively into the German-Austrian multicenter trial DAL-HD-90 for pediatric Hodgkin's disease (HD) in order to introduce local radiotherapy according to the individual patient's spread of disease within a combined-modality treatment. This paper investigates the feasibility of such a procedure and its impact on the final definition of the extent and stage of disease as well as on the choice of treatment. PATIENTS AND METHODS: Between October 1990 and July 1995, 578 children and adolescents <18 years (259 girls, 319 boys, median age 12.9 years) with HD were enrolled into the HD-90 trial. After clinical and pathological staging (66.4/33.6%), patients were allocated to treatment groups (TG) 1 'early stage', TG2 'intermediate stage', or TG3 'advanced stage'. All groups underwent two cycles of OPPA (vincristine, prednisone, procarbazine, doxorubicin) (girls) or OEPA (E, etoposide) (boys) for induction chemotherapy. TG2 and TG3 continued on as two or four cycles, respectively, of COPP (C, cyclophosphamide). Low-dose local radiotherapy was given to the initially involved sites, with radiation doses of 25 Gy in TG1/TG2, and 20 Gy in TG3. All documentation forms, radiographs, and chest and abdominal computed tomography (CT) scans were centrally reviewed, addressing in particular the individual patient's extent and stage of disease. This review and the set-up of individualized treatment proposals were in the hands of the study coordinator, one additional pediatrician and two radiation oncologists and radiologists at the study center. During a time slot of at least 8 weeks (two cycles of standard chemotherapy in all three TGs) the individualized treatment proposals were to be sent to the participating hospital. RESULTS: Complete sets of documentation from 564/578 patients (97.6%) were submitted sufficiently early to the study center. A total of 285 out of 574 chest radiographs, 468 out of 553 chest CT scans and 421 out of 548 abdominal CT scans were available from 71 hospitals. A total of 564 individualized treatment proposals were worked out by the review group and sent to the hospitals before radiotherapy began. Re-analysis of images and documentation forms, including laboratory and clinical data, resulted in a revision of stage in 115/571 patients (20.1%) and of TG in 76/571 patients (13.3%). A total of 67/76 patients were shifted into a higher TG, 60 patients on account of additionally detected extralymphatic involvement, five patients because of additionally detected lymph node involvement and two patients due to clinical data which had to be classified as B-symptoms. A total of 9/76 patients were shifted into a lower TG; in three patients extranodal disease and in six patents local lymph node involvement could not be confirmed. CONCLUSIONS: The up-front centralized review of patient data and consecutive set-up and delivery of individualized treatment proposals for almost every patient are feasible within a large multicenter trial. Sufficient time and manpower at the study center are needed for the review process and the set-up of individualized treatment proposals. Such a procedure has a significant impact on the homogeneity of stage definition, allocation to TG, and individualized treatment proposals. PMID- 11937247 TI - Intraoperative presacral electron boost following preoperative chemoradiation in T3-4Nx rectal cancer: initial local effects and clinical outcome analysis. AB - BACKGROUND AND PURPOSE: To analyze early results of a single institution experience using adjuvant intraoperative electron radiation therapy (IOERT) presacral boost in locally advanced rectal cancer following preoperative chemoradiation. MATERIALS AND METHODS: In a 63 month period (March 1995-June 2000), 100 consecutive T(3-4)N(x) rectal cancer patients were treated with preoperative chemoradiation (45-50 Gy plus oral Tegafur or 5-Fluorouracil continuous intravenous infusion), radical surgery and IOERT presacral boost (mean dose, 12.5 Gy; range, 10-15 Gy). Adjuvant chemotherapy (5-FU-leucovorin: 4-6 cycles) was given to 52 patients. The median age was 63 years, and 39 patients were >or=70 years old (65 males). Clinical staging was performed with computed tomography (94%) and/or endorectal ultrasound (71%) categorizing 90 cT(3), 10 cT(4), 20 cN(x), and 36 cN(+). Abdomino-perineal resection was performed in 41 cases. RESULTS: The IOERT cancellation rate was 6%. With a median follow-up of 23 months in IOERT treated patients, three developed pelvic recurrence: one anastomotic and one in the posterior vaginal wall (simultaneously with distant metastatic disease); and one presacral (in-field IOERT) as the only site of initial failure. Distant metastasis has been observed in 14 patients (exceptionally in pT(0-1) downstaged patients: 1/20; 5%). Overall treatment tolerances, including neoadjuvant and surgical segments, were acceptable. The actuarial 4-year estimations of local control, disease-free and overall survival are 94, 75 and 65%, respectively. CONCLUSIONS: IOERT electron boost to the presacral region is feasible to integrate systematically in the intensive combined treatment of locally advanced rectal cancer, including neoadjuvant chemoradiation segment. Topography of pelvic recurrences identified 2/3 relapses located in non-IOERT boosted anatomic intrapelvic sites: posterior vaginal wall and anastomotic suture. Presacral recurrence in locally advanced rectal cancer seems to be of low incidence, in a non-subspecialized academic surgical practice coordinated with a multidisciplinary oncology evaluation context, if an IOERT boost is included as a component of treatment together with preoperative chemoradiation. PMID- 11937248 TI - Preoperative irradiation and surgery for recurrent rectal cancer. Will intraoperative radiotherapy (IORT) be of additional benefit? A prospective study. AB - BACKGROUND: The therapeutic gain of surgery for recurrent rectal cancer is not clear, particularly with regard to the addition of intraoperative radiotherapy (IORT). METHODS: Patients (107) with isolated pelvic recurrence of rectal cancer received preoperative external radiotherapy of 46-50 in 2 Gy fractions. At surgery 59 patients had IORT 12-18 Gy. Survival and local recurrence was analysed with regard to surgical resection stages and IORT. RESULTS: Patients (44) had R0- and 39 R1-resections, 24 R2-resections or exploratory laparotomy. IORT was given most often after R1-resections, least in R0-patients. Estimated 5-year survival was overall around 30%, around 60% in the R0-, around 25% for R1- and 0% in R2 patients. Local recurrence was around 30% in the R0- and around 65% in R1-stage patients. R0-/R1-stage patients survived statistically significantly longer than the R2-group otherwise there was no statistical significant difference between IORT and non-IORT groups in any R-stages regarding overall survival or local recurrence. CONCLUSIONS: Macroscopic removal of the recurrence improves survival. Whether R0- is better than R1-resections is not clear. The effect of IORT is not a major one. IORT need be evaluated in randomised controlled trials. PMID- 11937249 TI - Distressful symptoms after radical radiotherapy for urinary bladder cancer. AB - BACKGROUND: Radical radiotherapy for muscle-invasive urinary bladder cancer can sterilize the tumour with preserved organ function. Here we studied symptoms, symptom distress and trade-off among long-term survivors and compared figures to those of population controls and patients who had undergone cystectomy. MATERIALS AND METHODS: We identified 71 patients who had had urinary bladder cancer treated with radical radiotherapy before 1995. For comparison, 325 patients treated with radical cystectomy and urostomy, continent or non-continent, during the same period and 460 individuals randomly selected from the general population were included. Information was collected by means of an anonymously answered postal questionnaire to avoid investigator-related bias. RESULTS: Answers were obtained from 58 (82%) radiated patients, 251 (85%) cystectomized patients and 310 (71%) population controls. Of the radiated patients, 74% reported little or no distress from symptoms from the urinary tract, 38% had had intercourse the previous month and 57% (men) reported they had ejaculated. Among the cystectomized patients, 13% had had intercourse and 0% (men) had ejaculated. Moderate or much distress from symptoms from the gastrointestinal tract was reported by 32% of the radiated patients, 24% of the cystectomized patients and 9% of the population controls. After radical radiotherapy, 46% of the patients were willing to accept some risk of decreased survival to become symptom-free. CONCLUSIONS: About 3/4 of these long-term survivors after radical radiotherapy for bladder cancer had a functioning urinary bladder with little or no distress from the urinary tract. The prevalence of sexual dysfunction was lower than after cystectomy and the prevalence of distress from the gastrointestinal tract was comparable. PMID- 11937250 TI - A cost-minimising analysis of standard radiotherapy and two experimental therapies in glioblastoma. AB - BACKGROUND AND PURPOSE: Accelerated radiotherapy (ART) and intracavity brachytherapy (ICBT) have been introduced in the primary treatment of glioblastoma. Our objective was to determine total treatment costs, hospitalisation time, and treatment outcome in these two experimental therapies compared to standard treatment. MATERIALS AND METHODS: In the time period 1985 to 1st May 1999, a total of 174 patients with histologically confirmed glioblastoma multiforme were given postoperative radiotherapy according to three different treatment schedules at three different time intervals. A conventional regime of external radiotherapy (54Gy/30 fractions) was given to 58 patients (group I), 75 patients were treated with ART (48Gy/twice daily 30 fractions) (group II), and 41 patients were given ICBT (60Gy/ten fractions) (group III). Treatment costs including surgery, hospital stay, hospital hotel stay, and radiotherapy were calculated. RESULTS: The total mean costs employing the three treatment alternatives were calculated to $25,618 (group I), $23,442 (group II), and $14,534 (group III). Total mean stay in hospital for the whole primary treatment was 48.8, 41.6, and 19 days for groups I, II, and III respectively. Median survival figures were 16, 14, and 13 months for groups I, II, and III, respectively. CONCLUSIONS: The total cost of postoperative radiotherapy in glioblastoma is comparable to other health care services. ART did not improve the total treatment cost or influence the need for hospitalisation compared to standard treatment. ICBT seemed to have economic benefits with less need for hospitalisation. PMID- 11937251 TI - Radiosurgery for brain metastases: the Tuebingen experience. AB - PURPOSE: To retrospectively investigate the effectiveness of linear accelerator based radiosurgery (RS) in the treatment of brain metastases (BM). MATERIAL AND METHODS: Of 55 patients with a total of 72 BM, 41 patients had a single brain metastasis and 14 patients had two or three metastases. Median tumour dose of 15Gy (range 8-20Gy) was prescribed to a median isodose surface of 90% (range 70 100%) encompassing the target volume. RESULTS: The median survival time (MST) for all 55 patients was 7 months [95% confidence interval (CI), 5-10 months] and 2 year survival is 18%. There was no significant difference between patients who had one brain metastasis and those with either two or three metastases (log rank P=0.7565). Multivariate analysis in patients with a single BM showed that interval between primary diagnosis (PD) to BM, maximum size of metastasis, and histology (renal cell carcinoma and melanoma versus others) were independent prognostic factors influencing survival. Local control was obtained in 66/72 (92%) metastases. Actuarial local control at 24 months was 52%. Only age (50 years) and histology (renal cell versus others) influenced local control in the univariate analysis in patients with a single BM. In multivariate analysis, size, histology (renal cell and melanoma versus others), activity of extracranial metastatic disease, age, interval from PD to BM and location (midline versus other) independently influenced local control, while the dose was not significant for our patient group. Only one patient developed radiographically suspected RS-induced necrosis after previous whole brain RT. CONCLUSION: RS was effective and little toxic in BM. Identification of prognostic factors must be performed to gain knowledge on patients most likely to benefit from this procedure. PMID- 11937252 TI - Changes in bone mineral density of lumbar spine after pelvic radiotherapy. AB - Bone mineral densities (BMDs) of L2 to L5 were measured on 40 cervical cancer patients with radiotherapy and 40 matched controls. We found no significant difference in the BMDs between the two groups and no significant change in BMDs 1 7 years after the therapy in the patient group. PMID- 11937253 TI - No correlation between endoscopic and symptomatic scoring systems after the treatment of radiation esophagitis. PMID- 11937254 TI - Electromyography and ratings of lumbar muscle fatigue using a four-level staircase protocol. AB - OBJECTIVE: The purpose of the study was to explore whether a relationship existed between subject's experience and objective measurements of back-muscle fatigue in healthy subjects. This may be used as reference material later. DESIGN: Muscle fatigue in the lower back was measured in healthy subjects using a staircase protocol. BACKGROUND: Muscle fatigue measurement in the lower back is of importance when assessing patients with low-back pain. The subject's experience of muscle fatigue needs to be explored further. METHODS: Twenty healthy subjects with informed consent participated. Borg CR-10 scale responses and electromyography measurements were obtained from low-back muscles of the subjects following a staircase protocol with sustained contractions at 20%, 40%, 60%, and 80% of maximum voluntary contraction. RESULTS: A strong correlation was found between Borg ratings and force (r=0.83) and between slope and force (r= -0.72) but not between Borg ratings and slopes which was at best 0.43 (/r/< or =0.43). CONCLUSIONS: Although subjective ratings and slopes showed a low correlation; increases in force with a concomitant increase in fatigue was reflected both by the subjective ratings and the slope. One cannot use subjective ratings for prediction of the slope for this type of protocol. However, a more complete description of muscle fatigue needs both the slope and the Borg ratings. RELEVANCE: In clinical testing, both subjective and objective measures of muscle fatigue must be considered since they measure different aspects of muscle fatigue. The median frequency slope probably reflects "physical discomfort" but the Borg ratings add a factor of "lack of energy". PMID- 11937255 TI - Abdominal and erector spinae muscle activity during gait: the use of cluster analysis to identify patterns of activity. AB - OBJECTIVE: To describe patterns of muscle activation during gait in selected abdominal and lumbar muscles using cluster analysis. PARTICIPANTS: A sample of convenience of 38 healthy adult volunteers. Outcome measures. Electromyographic activity from the right internal and external obliques, rectus abdominis and lumbar erector spinae were recorded, and the root mean square values for each muscle were calculated throughout the stride in 5% epochs. These values were normalised to maximum effort isometric muscle contractions. Cluster analysis was used to identify groups of subjects with similar patterns of activity and activation levels. RESULTS: Cluster analysis identified two patterns of activity for the internal oblique, external oblique and rectus abdominis muscles. In the lumbar erector spinae, three patterns of activity were observed. In most instances, the patterns observed for each muscle differed in the magnitude of the activation levels. In rectus abdominis and external oblique muscles, the majority of subjects had low levels of activity (<5.0% of a maximum voluntary contraction) that were relatively constant throughout the stride cycle. In the internal oblique and the erector spinae muscles, more distinct bursts of activity were observed, most often close to foot-strike. The different algorithms used for the cluster analysis yielded similar results and a discriminant function analysis provided further evidence to support the patterns observed. CONCLUSIONS: Cluster analysis was useful in grouping subjects who had similar patterns of muscle activity. It provided evidence that there were subgroups that might otherwise not be observed if a group ensemble was presented as the "norm" for any particular muscle's role during gait. RELEVANCE: The identification of common variations in muscle activity may prove valuable in identifying individuals with electromyographic patterns that might influence their chances of sustaining injury. Alternatively, clusters may provide important information related to muscle activity in those that do well or otherwise after a particular injury. PMID- 11937256 TI - Force-deformation response of the lumbar spine: a sagittal plane model of posteroanterior manipulation and mobilization. AB - OBJECTIVE: To develop a mathematical model capable of describing the static and dynamic motion response of the lumbar spine to posteroanterior forces. DESIGN: Static, impulsive and oscillatory forces with varying thrust angles and offsets were applied to the model, and the resulting motion responses were compared to experimental data published for spinal mobilization and manipulation of prone lying subjects. BACKGROUND: Methods are sought to improve understanding of the dynamic force-induced displacement response of the lumbar spine during spinal mobilization and manipulation treatment. METHODS: The thorax, pelvis and five lumbar vertebrae were represented as seven rigid structures and eight flexible joint structures. Flexible joint structures were modeled using spring and damper elements with three displacement degrees-of-freedom (posterior-anterior and axial displacement, and flexion-extension rotation). The resulting 21 degrees-of freedom lumped parameter model was solved in modal space. RESULTS: The fundamental natural frequency of vibration was 5.24 Hz. Simulations performed using 100 N static and dynamic posteroanterior forces applied to the L3 vertebrae indicated that peak L3 segmental displacements were up to 2.40 mm (impulsive) and 8.23 mm (oscillatory at 2 Hz). Appreciable axial displacements (0.41 mm) and flexion-extension rotations (1.49 degrees ) were also observed for oscillatory forces at L3. The posteroanterior motion response of the lumbar vertebrae was relatively insensitive to both the thrust force angle and thrust force offset, but axial displacements and flexion-extension rotations showed a large change (2 fold or greater) for thrust angles greater than -5 degrees (caudal) in comparison to vertical thrusts. Intersegmental motion responses for static, impulsive and oscillatory loads were more comparable than their segmental counterparts. CONCLUSIONS: The model predicts lumbar segmental and inter-segmental motion responses to manipulative forces that are otherwise difficult to obtain experimentally. RELEVANCE: This study assists clinicians to understand the biomechanics of posteroanterior forces applied to the lumbar spine of prone-lying subjects. Of particular clinical relevance is the finding that greater spinal mobility is possible by targeting specific load-time histories. PMID- 11937257 TI - Force measurements of postural sway and rapid arm lift in seated children with and without MMC. AB - OBJECTIVE: The aim was to investigate the horizontal ground reaction forces of seated postural sway and rapid arm lift in children with and without myelomeningocele. BACKGROUND; It is unclear whether children with myelomeningocele have limited control of body posture entirely caused by the impairment in the legs or also by other dysfunction. METHODS: 11 children with myelomeningocele, 10-13 years, and 20 children without physical impairment were investigated. Data were collected by force plate measurements during quiet sitting and during rapid arm lift. The forces were expressed as the corresponding acceleration of the centre of mass. The amplitude and the frequency of the centre of mass acceleration quantified the sway. Movement time, onset and anteroposterior peak acceleration were analysed during arm lift. RESULTS: The children with myelomeningocele had a low sway frequency under both conditions: eyes open and eyes closed. The movement time was longer for these children compared to the controls. The onset of initial anteroposterior centre of mass acceleration preceded the arm lift and was directed forward in both groups. The peak centre of mass acceleration was usually directed backward. CONCLUSIONS: The control of postural sway was different in children with myelomeningocele compared to children without disabilities and this could not be explained by the cele level. The children with myelomeningocele had a slow motor performance of the seated sway and during arm lift. RELEVANCE: Slow motor performance involves functional limitations in the individual child and is important for the therapy program. PMID- 11937258 TI - Analysis of postural control synergies during quiet standing in healthy children and children with cerebral palsy. AB - OBJECTIVE: To estimate the contribution of body transverse rotation using weighted differential center of pressure signals during quiet standing in healthy children and in children with cerebral palsy. DESIGN: Body sway was indirectly measured through center of pressure data, which was calculated using dual force platforms. BACKGROUND: Assessment of postural control synergies using center of pressure data provides a unique method for center of mass data analysis in characterizing complex balance sway. METHODS: Using dual force platforms, linear expressions for the coordinates of right and left center of pressure signals were developed to identify and characterize balance control synergies during quiet standing. Subjects were also tested during eyes open and eyes closed trials to determine the significance of visual input on these control synergies. RESULTS: The limb protraction/retraction control was found to be dominant during medial lateral sway, whereas the estimated body transverse rotation contribution was found to be more significant than the previously reported measures of anterior posterior balance. These findings were consistent in healthy children and in children with cerebral palsy during both eyes open and eyes closed trials. CONCLUSION: The weighted differential center of pressure signals show that the estimated body transverse rotation contribution is significant in healthy children and critical for postural stability in children with cerebral palsy. RELEVANCE: This study identifies the significance of body transverse rotation control contribution in upright posture. Children with cerebral palsy with relatively poor ankle control demonstrate the importance of body transverse rotation for postural stability. PMID- 11937259 TI - Load on the shoulder in low intensity wheelchair propulsion. AB - OBJECTIVE: To assess the mechanical load on the glenohumeral joint and on shoulder muscles during wheelchair propulsion at everyday intensities. DESIGN: Model simulations based on experimental input dataBackground. Virtually nothing is known about the mechanical load on the upper extremity during wheelchair propulsion. Hand rim wheelchair propulsion is a significant risk factor for shoulder pain and injury among wheelchair users. A musculoskeletal model of the upper extremity during wheelchair propulsion will quantify the stresses placed on anatomic structures and may provide insight into the source of symptoms and injuries. METHODS: Three experienced wheelchair users underwent wheelchair exercise tests at combinations of two load levels (10 and 20 W) and two velocities (0.83 and 1.39m.s(-1)) during which input data were collected for a musculoskeletal model of the upper extremity. The model was then used for the estimation of the glenohumeral contact force, as well as individual muscle forces. RESULTS: Peak glenohumeral contact forces were between 800 and 1400 N (100-165% body weight) and differed significantly between load levels. Averaged over the push phase, these forces were 500-850 N. In absolute terms the m. deltoideus and rotator cuff muscles were highly active (>100N). In relative terms the load on the m. supraspinatus was high, with peak values of over 50% of its maximum attainable force. CONCLUSIONS: Low intensity wheelchair propulsion does not appear to lead to high contact forces. The muscle forces in the rotator cuff and especially in the m. supraspinatus are high. This might indicate a risk for muscle damage and the subsequent development of shoulder complaints, such as rotator cuff tears. RELEVANCE: Within the wheelchair user population, there is a high prevalence of upper extremity complaints. Not much is known about the causes of those complaints. Wheelchair propulsion is likely to be a major risk factor. If the (nature of this) mechanical load can be identified, specific exercise programs and/or design changes can be better tuned to prevent overuse injuries. PMID- 11937260 TI - Consequence of feedback-based learning of an effective hand rim wheelchair force production on mechanical efficiency. AB - OBJECTIVE: Investigation of the effect of visual feedback on effective hand rim wheelchair force production and the subsequent effect on gross mechanical efficiency. DESIGN: Ten subjects in an experimental group and 10 subjects in a control group practised three weeks (3.wk(-1), i.e., a pre-test and 8 trials) on a computer-controlled wheelchair ergometer. Every trial consisted of two blocks of 4 min at 0.15 and 0.25 W.kg(-1) at 1.11 m.s(-1). On three trials an additional block at 0.40 W.kg(-1) was performed. The experimental group practised with and the control group practised without visual feedback on the effectiveness of force production. BACKGROUND: In mechanical terms, the low gross mechanical efficiency of hand rim wheelchair propulsion may be the result of ineffective force production. METHODS: During all trials oxygen uptake, power output, forces and torque on the hand rims were measured. RESULTS: In comparison with the control group, the experimental group at trial 8 had a significantly more effective force production compared to the control group (90-97% vs. 79-83%, respectively), but showed a significantly lower mechanical efficiency (5.5-8.5% vs. 5.9-9.9%, respectively). CONCLUSION: Findings indicate that the most effective force production from a mechanical point of view is not necessarily the most efficient way--in terms of energy cost-- from a biological point of view and that force direction is based on an optimization of cost and effect. RELEVANCE: Learning a more effective force production by visual feedback is not useful for increasing the mechanical efficiency of hand rim wheelchair propulsion. PMID- 11937261 TI - Gait adaptations in patients with chronic posterior instability of the knee. AB - OBJECTIVE: A retrospective analysis was performed to assess gait in individuals with a long history of posterior knee instability. DESIGN: Descriptive study. BACKGROUND: There are few studies in the literature concerning evaluation of the biomechanics of the knee in patients with knee posterior instability. METHODS: Nine individuals with posterior knee instability and a matched control group of uninjured subjects were tested in regards to knee kinematics and kinetics while walking and ascending and descending stairs. The mean follow up time for the individuals with posterior instability was 11.1 years. Individual satisfaction with the knee was measured by having participants complete the Flandry (also known as Hughston Clinic) self-assessment questionnaire. RESULTS: It was found that patients with knee posterior instability who indicated a higher level of satisfaction on the Flandry score walked in a manner that demonstrated greater peak knee extensor torque during stance phase, while less satisfied patients with knee posterior instability demonstrated lower peak knee extensor torque. There was a significant correlation between the self-assessment score and the peak knee extensor torque during level walking (P=0.003). During stair ascent and descent, patients with posterior instability averaged lower knee extensor torque and power than the control subjects, but those differences were only statistically significant in power while descending stairs (P=0.048). CONCLUSIONS: Individuals with chronic knee posterior instability modify their gait, and the adaptation can be predicted based upon the individuals self-assessment of their knee using the Flandry questionnaire. RELEVANCE: These data suggest that gait retraining may be a valuable addition to the traditional muscle strengthening programs, which are commonly used during conservative management of knee posterior instability. PMID- 11937262 TI - NFkappaB-dependent signaling pathways. AB - The transcription factor NFkappaB is activated by numerous stimuli. Once NFkappaB is fully activated, it participates in the regulation of various target genes in different cells to exert its biological functions. NFkappaB has often been referred to as a central mediator of the immune response, since a large variety of bacteria and viruses can lead to the activation of NFkappaB, which in turn controls the expression of many inflammatory cytokines, chemokines, immune receptors, and cell surface adhesion molecules. Recent studies have shown that NFkappaB may function more generally as a central regulator of stress responses, since different stressful conditions, including physical stress, oxidative stress, and exposure to certain chemicals, also lead to NFkappaB activation. Furthermore, NFkappaB blocks cell apoptosis in several cell types. Taken together, these findings make it clear that NFkappaB plays an important role in cell proliferation and differentiation. It is the intention of this review to cover the various NFkappaB-dependent signaling pathways, thereby to achieve a better understanding of the mechanisms of NFkappaB activation and the physiological functions of activated NFkappaB. PMID- 11937263 TI - Keratinocyte growth factor regulates proliferation and differentiation of hematopoietic cells expressing the receptor gene K-sam. AB - OBJECTIVE: The aim of this study was to establish a new method to overcome the problems of gene therapy targeting hematopoietic cells, namely low transduction efficiency and induction of differentiation during cytokine treatment. MATERIALS AND METHODS: The K-sam gene encoding the receptor for keratinocyte growth factor (KGF) was transduced to three factor-dependent hematopoietic cell lines (Ba/F3, 32Dcl3, and UT-7/GM) using retroviral vector, and their proliferation, differentiation, and intracellular signaling were studied. This gene also was transduced to murine bone marrow cells, and proliferation of colony-forming cells (CFCs) by KGF stimulation was examined. RESULTS: Although KGF is known to target only epithelial cells, all of the three cell lines transduced with K-sam proliferated due to KGF stimulation. Morphologic observation showed that KGF induced proliferation but did not cause significant differentiation of 32D/K-sam cells. KGF treatment increased phosphorylation of ERK1/2 but did not activate STAT molecules. Granulocyte colony-stimulating factor transduced the differentiation signal with the phosphorylation of STAT3 without significant ERK1/2 activation. Proliferation by KGF of murine primary bone marrow cells transduced with K-sam then was examined in liquid culture. KGF treatment significantly increased production of CFCs derived from K-sam-transduced bone marrow cells without causing the exhaustion of immature CFCs. CONCLUSIONS: KGF could efficiently induce proliferation of hematopoietic cells expressing the K sam gene without obvious induction of differentiation or exhaustion of immature progenitor cells. The in vitro data are important for further preclinical in vivo study. PMID- 11937264 TI - Regulation and function of the CGRP receptor complex in human granulopoiesis. AB - OBJECTIVE: Anatomic studies and animal experiments suggest that neuropeptides such as calcitonin gene-related peptide (CGRP) may be involved in hematopoiesis. Here, we examined the regulation and function of the CGRP receptor in human granulopoiesis. MATERIALS AND METHODS: Expression of CGRP receptor components on CD34(+) cells, peripheral blood granulocytes, and in vitro differentiated CD34(+) cells was analyzed at the mRNA level and by measuring the signaling capacity of the receptor. The function of CGRP in human hematopoiesis was investigated by clonal colony formation assays. RESULTS: mRNA transcripts for the cell surface CGRP receptor subunits receptor activity-modifying protein-1 (RAMP-1) and calcitonin receptor-like receptor (CRLR) as well as for the intracellular adapter protein CGRP-receptor component protein (CGRP-RCP) were found in CD34(+) cells from 4/4 donors tested. CGRP-RCP mRNA was expressed in peripheral blood granulocytes of 8/15 donors, whereas RAMP-1 and CRLR were not detectable. CD34(+) cells, but not granulocytes, exhibited a marked elevation of cellular cAMP after CGRP stimulation, thereby confirming the mRNA expression data. Both RAMP-1 and CRLR mRNA expression and CGRP receptor signaling capacity were lost during in vitro granulocytic differentiation of CD34(+) cells. Consistent with a role of CGRP in hematopoiesis, we show that CGRP significantly enhances the formation of granulomonocytic, but not erythroid or mixed, colonies by purified human CD34(+) cells. CONCLUSION: The CGRP receptor is expressed on CD34(+) hematopoietic progenitor cells and is downregulated during granulocytic differentiation. CGRP directly acts on CD34(+) cells to promote formation of granulomonocytic colonies. Thus, CGRP may have a function in directing hematopoiesis. PMID- 11937265 TI - Overexpression of novel short isoforms of Helios in a patient with T-cell acute lymphoblastic leukemia. AB - OBJECTIVE: In previous studies, we demonstrated overexpression of the dominant negative isoform of the transcription factor Ikaros, Ik-6, in patients with blast crisis of chronic myelogenous leukemia and B-cell acute lymphoblastic leukemia. In the present study, we analyzed expression of the Ikaros family genes Ikaros, Aiolos, and Helios in a panel of human T-cell leukemia/lymphoma cell lines and bone marrow samples of patients with T-cell acute lymphoblastic leukemia. MATERIALS AND METHODS: We performed reverse transcriptase polymerase chain reaction, sequencing analysis, immunoblotting, and Southern blotting. RESULTS: We found overexpression of novel short isoforms of Helios (Hel-5 and Hel-6) in the HD-Mar cell line. Southern blot analysis suggested that there might be a small deletion in the Helios locus of HD-Mar. In addition, we observed decreased expression of more than one Ikaros family gene in 3 of 9 patients with T-cell acute lymphoblastic leukemia. Moreover, one of the patients overexpressed novel short isoforms of Helios (Hel-7 and Hel-8). CONCLUSION: This study provides the first evidence of an Ikaros family member (other than Ikaros) of which novel short isoforms become overexpressed in human leukemia. PMID- 11937266 TI - Loss of genomic imprinting of insulin-like growth factor 2 is strongly associated with cellular proliferation in normal hematopoietic cells. AB - OBJECTIVE: The human insulin-like growth factor 2 (IGF2) gene was thought to be imprinted and expressed only from the paternal allele in normal tissue. MATERIALS AND METHODS: Initially, we analyzed the imprinting status of IGF2 in bone marrow cells from 49 patients with myelodysplastic syndromes (MDS) utilizing the Apa I polymorphism of IGF2. Thirteen bone marrow and 14 peripheral blood samples from normal individuals served as controls. We utilized normal peripheral blood T lymphocytes to examine the relationship between genomic imprinting and cell proliferation. Expression of IGF2 was quantified by real-time PCR and proliferation of T cells was measured by 3H-thymidine incorporation. Furthermore, methylation status of the imprinting controlling region (ICR) was analyzed by subcloning and sequencing of genomic DNA after sodium bisulfite modification. RESULTS: Among 24 patients who were heterozygous for IGF2, loss of imprinting (LOI) occurred in 22 cases (92%). Surprisingly, LOI of IGF2 occurred in the normal bone marrow cells, but the normal peripheral blood cells showed retention of imprinting (ROI). Unstimulated normal T cells showed ROI. After 24 hours of exposure to PHA, these cells changed their IGF2 imprinting status from ROI to LOI. Concomitantly, their IGF2 RNA levels increased up to sixfold and their proliferation increased 10- to 20-fold. In contrast, normal T cells not stimulated with PHA did not develop LOI of IGF2, had negligible levels of IGF2 RNA, and did not increase their proliferation. In unstimulated T cells, the CpG islands of the ICR were completely methylated on one allele and nearly completely unmethylated on the other allele. After PHA stimulation, the CpG islands at the ICR became completely methylated on both alleles. CONCLUSION: LOI of IGF2 is strongly associated with cell proliferation and is not limited to cancer cells. PMID- 11937267 TI - Contact with fibronectin enhances preservation of normal but not chronic myelogenous leukemia primitive hematopoietic progenitors. AB - OBJECTIVE: Coculture with stromal cells enhances preservation and self-renewal of primitive progenitor potential in hematopoietic cells during ex vivo culture with growth factors (GF). However, the respective roles of growth factors, stromal contact, and extracellular matrix (ECM) ligands in this effect are not clear. Here we investigated the role of direct contact with stroma and the ECM protein fibronectin (FN) in these effects, and investigated whether abnormal integrin receptor function in chronic myelogenous leukemia (CML) progenitors was associated with perturbation in these responses. METHODS: Normal bone marrow CD34+ cells were cultured in GF-containing medium with or without contact with stromal layers, glutaraldehyde-fixed stromal layers (stroma-contact), or integrin binding FN fragments for 7 days. Progeny cells were assayed for primitive progenitors in week-6 long-term culture-initiating cell (LTC-IC) and week-10 extended LTC-IC (ELTC-IC) assays. RESULTS: Increased LTC-IC and ELTC-IC preservation was seen following coculture with stroma, and was also observed after culture in contact with fixed stromal layers and FN. Both alpha4beta1 and alpha5beta1-integrin binding FN fragments enhanced LTC-IC preservation. Analysis of single CD34+CD38- cells showed that coculture with FN resulted in significantly reduced cell division, but enhanced retention of LTC-IC capacity in divided cells. FN also increased LTC-IC frequency in undivided cells. CML progenitors demonstrate deficient integrin-mediated adhesion, migration, and signaling. Coculture of CML CD34+ cells with stroma and FN failed to enhance LTC IC preservation. CONCLUSION: We conclude that beta1 integrin-FN interactions enhance normal primitive progenitor preservation with or without cell division, and that these mechanisms are impaired in CML primitive progenitors. PMID- 11937268 TI - Increased incidence of spontaneous apoptosis in the bone marrow of hyperdiploid childhood acute lymphoblastic leukemia. AB - OBJECTIVE: Hyperdiploidy of 51-65 chromosomes is associated with a good prognosis in childhood B-lineage acute lymphoblastic leukemia (ALL). Blasts from childhood ALL patients with a hyperdiploid karyotype have a tendency to apoptosis when cultured on stromal layers in vitro. In this study, we apply a novel method to investigate the relationship between apoptosis and hyperdiploidy in lymphoblasts of childhood ALL. MATERIALS AND METHODS: The DNA content of individual ALL blasts in Feulgen-stained archival bone marrow smears can be determined by static cytometry. TUNEL (TdT-mediated dUTP-biotin nick end labeling) detects the DNA degradation associated with apoptosis. We performed TUNEL in situ sequential to DNA ploidy analysis in archival bone marrow smears from 12 patients with childhood ALL. RESULTS: Five patients were diploid and seven were hyperdiploid (51-65 chromosomes) by conventional cytogenetic analysis. In the five diploid cases, the percentage of TUNEL-positive blasts ranged from 1.0% to 1.3%; in the seven hyperdiploid cases, the percentage of TUNEL-positive blasts ranged from 3.6% to 9.0%. Comparing TUNEL and corresponding Feulgen images, we found that apoptotic blasts were predominantly of high DNA ploidy in both diploid and hyperdiploid cases. The mean DNA value of apoptotic blasts was larger than that of the total blast population in each case. CONCLUSIONS: The results demonstrate an increased incidence of spontaneous apoptosis in situ of hyperdiploid blasts in ALL bone marrow and indicate that this phenomenon is not restricted to in vitro cultures. The findings provide a possible rationale for the good prognosis associated with hyperdiploid childhood ALL. PMID- 11937269 TI - Human T-cell lymphotropic virus type I Tax activates lung resistance-related protein expression in leukemic clones established from an adult T-cell leukemia patient. AB - OBJECTIVE: We examined the significance of human T-cell lymphotropic virus type I (HTLV-I) Tax protein-induced resistance to anticancer drugs and the relationship between Tax and multidrug resistance proteins. MATERIALS AND METHODS: S1T cell, a leukemic non-Tax-producing T-cell clone established from an adult T-cell leukemia (ATL) patient, S1TcTax05 and S1TcTax10 clones, transfected with Tax stably expressing cDNA, and S1Tneo, transfected with a neomycin-resistant gene, were examined for Tax-related anticancer drug resistance. Resistance of those cells to the anticancer drugs doxorubicin, etoposide, cisplatin, and vindesine was tested with the MTT method. Expression of multidrug resistance protein mRNAs (MDR1, MRP1, cMOAT/MRP2, and LRP) was analyzed with reverse transcriptase polymerase chain reaction (RT-PCR). Doxorubicin subcellular distribution in those cells was examined by fluorescence microscopy. RESULTS: S1TcTax05 and S1TcTax10 showed resistance to doxorubicin, etoposide, and vindesine, but not to cisplatin as compared with S1T or S1Tneo. RT-PCR demonstrated that MRP1 mRNA was expressed, but MDR1, cMOAT, and LRP mRNAs were not in S1T or S1Tneo. Marked expression of LRP mRNA was detected, but no change of MDR1, MRP1, or cMOAT mRNA expression in Tax-expressing S1TcTax05 and S1TcTax10. Fluorescence microscopy demonstrated that doxorubicin was distributed mainly in the cytoplasm of S1TcTax05 and S1TcTax10, and in the nucleus of S1T and S1Tneo. CONCLUSIONS: These findings suggest that Tax-related drug resistance of ATL cells is due to LRP and not MDR1, as reported previously. These findings in cells derived from an ATL patient suggest a novel mechanism for drug resistance in Tax-expressing ATL cells. PMID- 11937270 TI - Functional evaluation of ex vivo expanded cord blood lymphocytes: possible use for adoptive cellular immunotherapy. AB - OBJECTIVE: To examine the possibility of adoptive cellular immunotherapy such as donor lymphocyte infusion using ex vivo expanded cord blood (CBL) lymphocytes, the potential expansion ability of CBL lymphocytes and the function of expanded CBL lymphocytes were evaluated. MATERIALS AND METHODS: Mononuclear cell fractions derived from CBL or peripheral blood (PBL) were placed in anti-CD3 monoclonal antibody-coated flasks and cultured in the presence of recombinant human interleukin-2 for 4 days. Cells then were transferred to noncoated flasks and cultured for another 2 weeks. On day 14, polyclonality, cell surface markers, killer activity, and intracellular cytokine profiles were evaluated. RESULTS: Cells were polyclonally expanded. The differences in cumulative fold expansion on day 14 between CBL [1174 +/- 637 (292-1939), n = 6] and PBL [1247 +/- 568 (517 2328), n = 9] were not significant (p = 0.95). Phenotypic patterns of both expanded CBL and PBL were similar. CD4/CD8 ratio of expanded CBL appeared to remain greater than 1 on day 8. In contrast, that of expanded PBL became less than 1. In both cases, approximately 20% of cells had the CD3(+)CD8(+)CD56(+) phenotype. At an effector to target ratio (E/T) of 40:1, the natural killer activity of expanded CBL (64.5% +/- 10.8%, n = 9) was significantly higher than that of expanded PBL (48.3% +/- 16.8%, n = 9) (p < 0.01, Mann-Whitney U-test). However, there was no significant difference in lymphokine-activated killer activity between expanded CBL (45.3% +/- 25.2%, n = 7) and expanded PBL (67.2% +/ 12.3%, n = 7). Interferon-gamma-producing cells were dominant in both cases. CONCLUSIONS: It was feasible to achieve approximately 1000-fold expansion of CBL, and the phenotype and function of expanded CBLs were essentially equivalent to those of expanded PBL. This suggested that ex vivo expanded CBL may be applied to adoptive cellular immunotherapy such as donor lymphocyte infusion. PMID- 11937271 TI - Absence of GPIbalpha is responsible for aberrant membrane development during megakaryocyte maturation: ultrastructural study using a transgenic model. AB - OBJECTIVE: The glycoprotein Ib/IX/V complex (GPIb-IX-V) mediates platelet attachment to von Willebrand factor in exposed subendothelium. Molecular defects in the genes for GPIbalpha, GPIbbeta, and GPIX give rise to the Bernard-Soulier syndrome, in which thrombocytopenia and giant platelets suggest that this receptor also is involved in platelet production. To study how giant platelets are produced in vivo, we used a model of GPIbalpha-deficient mice (GPIbalpha(null)) and mice rescued with the human GPIbalpha transgene (GPIbalpha(null;hTg)). MATERIALS AND METHODS: Using electron microscopy and immunogold labeling, we examined megakaryocytopoiesis in the bone marrow of these mice and developed a method to quantify the membranes of megakaryocytes (MK) and proplatelets by computer analysis. RESULTS: Abnormal membrane development in the perinuclear zone was found in immature MK of GPIbalpha(null) mice. This led to a poorly developed demarcation membrane system and other ultrastructural changes. As a result, well-organized platelet territories were rarely seen within the cytoplasm of mature MK. Membrane quantification confirmed that MK of GPIbalpha(null) mice had a reduced internal membrane pool. Whereas these MK normally crossed the endothelial barrier, their migration was accompanied by the production of unusually large MK fragments or proplatelets in the vascular sinus with an approximately 50% decrease in internal membrane content compared to wild type. In the rescued GPIbalpha(null;hTg) model, GPIbalpha was normally localized in MK, and there was a total correction of the ultrastructural defects. CONCLUSIONS: GPIbalpha is essential for membrane development and distribution in maturing MK. Its absence leads to abnormal partitioning of the membrane systems and abnormal proplatelet production. PMID- 11937272 TI - Expression of lineage markers by CD34+ hematopoietic stem cells of adult mice. AB - OBJECTIVE: By using a murine transplantation model, we studied the relationship between CD34 expression and expression of CD4 and Mac-1 by hematopoietic stem cells of normal adult mice. MATERIALS AND METHODS: Cells from Ly-5.1 C57BL/6 mice were used as test cells and lethally irradiated Ly-5.2 mice were used as recipient mice. Peripheral blood was obtained 6 months posttransplantation to analyze engraftment of donor-derived cells. RESULTS: First, we determined that CD34- long-term reconstituting cells are CD4-, while some CD34+ stem cells express CD4. We then studied Mac-1 expression. Mac-1(-) and Mac-1(low) populations of both CD34- and CD34+ cells were capable of long-term reconstitution. Mac-1(high) population of CD34+ cells but not of CD34- cells also engrafted. CONCLUSIONS: These results strongly indicate that depletion of Mac 1(+) and CD4(+) cells in stem cell purification may inadvertently discard significant populations of CD34+ stem cells. Since positive selection based on CD34 expression is the current practice for purification of human stem cells, our studies may possess implications in the purification of human hematopoietic stem cells. PMID- 11937273 TI - Cells with hemopoietic potential residing in muscle are itinerant bone marrow derived cells. AB - OBJECTIVE: The nature of cells residing in muscle giving rise to hemopoietic colonies in vitro or hemopoietic reconstitution in vivo has been unclear. The goal of the present study was to characterize these cells and uncover their potential site of origin. MATERIALS AND METHODS: Cells prepared from muscle were characterized for surface antigens (CD45, CD34, c-kit, Sca-1, CD31, VCAM-1), for their in vitro clonogenic capacity and in vivo repopulation potential either as unpurified cells or sorted subsets (CD45(+), CD45(-)). The presence of bone marrow (BM)-derived cells in muscle of mice reconstituted with marked BM cells before and after cytokine-induced mobilization was also examined. RESULTS: Our data show: 1) The yield of CD45(+) cells is higher in muscle of neonates and young animals. Their composite phenotype does not favor contamination by blood. 2) The capacity of fresh muscle cell explants to give rise to colonies in vitro and hemopoietic reconstitution in vivo is associated with CD45(+) cells. 3) Irradiated recipients reconstituted with marked BM cells harbor marked BM-derived cells (CD45(+) or CD45(-)) in their muscle several months after transplant. 4) Cytokine-induced mobilization of transplanted animals modestly increases the yield of BM-derived cells recovered from muscle, unlike the yields from spleen, liver, or peripheral blood (PB). CONCLUSIONS: Our data suggest a reinterpretation of previously published conclusions: hemopoietic colonies derived from fresh muscle explants do not originate from transdifferentiated muscle cells, but from BM-derived cells residing in muscle; the hemopoietic reconstituting potential of muscle cells is likewise attributed to these cells. PMID- 11937274 TI - Culture conditions affect the ability of ex vivo expanded peripheral blood progenitor cells to accelerate hematopoietic recovery. AB - OBJECTIVE: The aim of this study was to evaluate the effect of various ex vivo expansion conditions on the cell products and their ability to accelerate hematopoietic recovery in patients undergoing stem cell transplantation. PATIENTS AND METHODS: Unselected peripheral blood progenitor cells (PBPCs) from 43 breast cancer patients were seeded into gas-permeable bags containing serum-free media, granulocyte colony-stimulating factor, stem cell factor, and pegylated megakaryocyte growth and development factor. Between 2 and 24 x 10(9) PBPCs were cultured at 1, 2, or 3 x 10(6) cells/mL for 9 to 14 days. The expanded PBPCs and cryopreserved PBPCs containing at least 5 x 10(6) CD34(+) cells/kg were infused following completion of high-dose chemotherapy. RESULTS: Increasing culture duration from 9 to 11-14 days significantly improved the expanded cell yield and fold expansion, whereas increasing PBPC seeding density above 10(6) cells/mL had the opposite effect. The expanded cell dose was the only culture variable that correlated significantly with the time to neutrophil recovery (r = -0.66; p = 0.0001). Study patients had significantly faster neutrophil (p < 0.0001) and platelet (p = 0.001) recovery, reduced red cell transfusions (p = 0.01), and shorter hospital stays (p < 0.0001) than historical controls. The most clinically efficacious expanded cell product was seeded with 10(10) PBPCs at 10(6) cells/mL, then cultured for 11 days. The six patients in that cohort experienced 2.8 +/- 1 days of absolute neutropenia and had neutrophil recovery 5.7 +/- 0.8 days post transplant; none had a neutropenic fever or required intravenous antibiotics. CONCLUSION: Unselected PBPCs expanded ex vivo significantly impact hematopoietic recovery following high-dose therapy. Optimization of the expansion conditions enhances the efficacy of this cell product. PMID- 11937275 TI - Visualization of urinary stones by 3-D ultrasound with surface rendering. AB - The purpose of this study was to evaluate the diagnostic value of 3-D ultrasound with surface rendering in stones located in different parts of the urinary tract. A total of 55 patients with urinary stones were examined by transabdominal 3-D ultrasound (US) with surface rendering of the stone. In calculi of the prevesical ureter, bladder and prostatic urethra, transrectal 3-D endosonography was performed as well. The radiographic shape and the chemical analysis of the stone were correlated to the sonographic findings. The localization of the stone was: renal 23, ureteral 26, vesical 5 and urethral 1. All stones could be identified by 3-D US. Surface rendering produced useful results in stones > 2 cm with a 3.5 MHz transabdominal scanning probe, and in all stones that were accessible with a 7.5-MHz transrectal scanning probe. In stones < 1 cm, endoUS with surface rendering provides more information on the structure of the stone than plain radiography, but there is no correlation between surface and chemical analysis. PMID- 11937276 TI - Sonographic evaluation of ACL rupture signs compared to arthroscopic findings in acutely injured knees. AB - Evaluation of ultrasound (US) examination for the diagnosis of an acute anterior cruciate ligament (ACL) rupture. We performed a prospective examination of 193 patients to analyze the three indirect criteria of ACL rupture: echo-poor space at the femoral insertion of the ACL, S-shaped course and thickening of the posterior cruciate ligament (PCL) and protrusion of the posterior fibrous capsule. The assumption that the echo-poor space at the femoral insertion has a great significance was confirmed in this study. Sensitivity for ACL rupture was 0.91 and specificity was 0.80, protrusion of the posterior fibrous capsule had a specificity of 0.77 and a sensitivity of 0.68. The use of both criteria increased the sensitivity of sonography to 0.98, but the specificity decreased to less than 0.50. We conclude that the US examination with its three indirect signs of ACL rupture has a high predictive value in the acute traumatized knee joint. PMID- 11937277 TI - Real-time spatial compound imaging in breast ultrasound. AB - To determine the role of real-time spatial compound imaging in breast ultrasound (US), 38 patients with a total of 50 benign changes (fibroadenomas, cysts, lactiferous duct dilatation) underwent both conventional B-mode US and real-time spatial compound imaging under standardized examination settings. Subsequently, images were reviewed independently by three readers experienced in breast US and evaluated according to a multistage scoring system with regard to the presence of artefacts, delineation of boundaries and depiction of internal structures. With significant reader concordance, real-time spatial compound imaging was found to produce speckle reduction with improvement of tissue differentiation, increased conspicuity of low-contrast lesions, enhanced delineation of capsular margins and ducts, and improved depiction of internal architecture of solid lesions, as well as clearer visualization of cystic contents due to clutter reduction. Preservation of central acoustic enhancement and lateral edge shadowing in cysts and fibroadenomas, however, was recorded better in conventional imaging. PMID- 11937278 TI - High-flow priapism: a combined interventional approach with angiography and colour Doppler. AB - Traumatic high-flow priapism caused by a pathologic influx from lacerated arteries to the cavernous bodies is usually treated by transcatheter arterial embolisation. The purpose of this study was to evaluate a combined interventional approach with intraoperative perineal colour Doppler ultrasound (US) while performing the embolisation procedure. Our aim was to reduce radiation exposure and application of iodinised contrast media. To achieve an optimal position of the angiography catheter directly before or in the fistula, instead of several x ray examinations with contrast media, a perineal colour Doppler US examination was performed while saline or US contrast media were injected through the catheter. The flow pattern allows the assessment of the occlusion of the fistula and the intact flow in the arteries that were not damaged. In four patients (unilateral fistula: 1, bilateral fistulas: 3) with traumatic high-flow priapism, this technique was performed combined with a conventional angiographic control of the embolisation. Seven embolisation sessions were performed. Only in one case, a dislocation of a microcoil required a second session. In all cases, the priapism disappeared immediately after the final session, leading to a restored erectile function. This combined approach may reduce the exposure to radiation and contrast media, especially in children, who often suffer from multiple fistulas and need more than one session. The procedure also prevents an occlusion of intact arteries that affects the penile perfusion and increases the risk of erectile dysfunction. PMID- 11937279 TI - Contrast-enhanced power Doppler sonography: assessment of revascularization flow in Legg-Calve-Perthes' disease. AB - A total of 26 children (26 affected, 25 control hips) with Legg-Calve-Perthes' disease (LCP) met the cooperation requirements for inclusion in the study using contrast-enhanced power Doppler sonography for assessment of changes in the revascularization flow. Sagittal sonograms were obtained before and at the peak effect of an intravenously injected microbubble-based contrast agent. Overall mean peak enhancement ratios were greater for children with later onset of disease (>5.8 years old) than for those with earlier onset (< or = 5.8) (p = 0.02). The use of ultrasound (US) contrast agents improved depiction of proximal femoral vascularity in all regions of the femoral head: epiphysis (n = 3, pre; n = 8, postcontrast), physis (n = 12, pre; n = 21, postcontrast) and metaphysis (n = 6, pre; n = 8, postcontrast) and resulted in a marked increase in the number of mean pixel intensity values within the physis (p = 0.02). In conclusion, contrast enhanced power Doppler sonography is an effective method for demonstration of changes from the revascularization process in LCP, particularly within the physis. PMID- 11937280 TI - Human cerebral perfusion analysis with ultrasound contrast agent constant infusion: a pilot study on healthy volunteers. AB - With ultrasound (US) contrast agent (UCA) continuous infusion providing a steady state, mean tissue microbubble velocity can be assessed by analyzing the reappearance rate after microbubble destruction with US energy (refill kinetics). In this study, we investigated this new approach for the assessment of human cerebral perfusion. A total of 12 healthy volunteers were investigated transtemporally with increasing pulsing intervals (250, 500, 750, 1000, 1250, 1500, 2000, 3000 and 4000 ms) and two UCA infusion rates (0.5 and 1.0 mL/min of Optison). Intensity vs. pulsing interval curves were analyzed using an exponential curve fit and parameters of the curve (plateau echo enhancement, A, representing the microbubble concentration within the interrogated tissue; rate constant, beta, which is related to blood flow and their product, F = Abeta) were compared. For 20/20 investigations being available for further analysis, it was possible to generate a typical exponential intensity vs. pulsing interval curve from the ipsilateral thalamus. The plateau echo enhancement A showed a significant (p = 0.02), and the beta as well as the F values displayed a nonsignificant (p = 0.06, both), increase with infusion rate. The qualitative analysis of beta and F parameter images displayed the most homogeneous visualisation of perfusion in the ipsilateral thalamus and main territory of the middle cerebral artery. In conclusion, it is possible to display the UCA refill kinetics in human cerebral microcirculation after microbubble destruction by transcranial US. Grey-scale harmonic imaging allows a quantitative approach to cerebral perfusion with a large interindividual variation of the parameters. PMID- 11937281 TI - The impact of theoretical errors on velocity estimation and accuracy of duplex grading of carotid stenosis. AB - Two potential errors in velocity estimation, Doppler angle misalignment and intrinsic spectral broadening (ISB), were determined and used to correct recorded blood velocities obtained from 20 patients (38 bifurcations). The recorded and corrected velocities were used to grade stenoses of greater than 70% using two duplex classification schemes. The first scheme used a peak systolic velocity (PSV) of > 250 cm/s in the internal carotid artery (ICA), and the second a PSV ratio of > 3.4 (ICA PSV/common carotid artery PSV). The "gold standard" was digital subtraction angiography (DSA). The maximum error in velocity estimation due to Doppler angle misalignment was 33 cm/s, but this did not alter sensitivity of stenosis detection. ISB correction caused a reduction in PSV that decreased the sensitivity of the PSV scheme from 65% to 45%. The PSV ratio classification was not affected by ISB errors. Centres using a PSV criterion for grading stenosis should use a fixed Doppler angle and should establish velocity thresholds in-house. PMID- 11937282 TI - Reliability and validity of day-to-day blood flow velocity reactivity in a single subject: an fTCD study. AB - To evaluate the within-subject variability of repeated task-induced blood flow velocity (BFV) change, we measured the haemodynamics of the anterior and middle cerebral arteries of the same volunteer on 20 consecutive working days during the performance of mental arithmetic with transcranial Doppler ultrasonography. BFV changes with the arithmetic condition were compared against a resting and a counting condition. Absolute BFV data showed equally increased velocities during rest and arithmetic activity as compared to counting. The findings suggest that the resting state might not be the ideal reference paradigm to evaluate task induced haemodynamic changes. Selecting the counting condition as the more valid baseline measure, we obtained relatively stable lateralisation indices for the MCA with 1 of 10 measurements outside the 95% confidence interval. Reproducibility was poorer in the ACA, with 3 outliers. A strategy that calculates the average of multiple short activation epochs within the same session appears to improve the reliability of the lateralisation index. PMID- 11937283 TI - Detection of Barrett's epithelium by acoustic microscopy. AB - Barrett's esophagus is associated with increased risk of adenocarcinoma of the gastroesophageal junctional region. The presence of goblet cells (intestinal metaplasia) in columnar cell-lined esophageal mucosa defines Barrett's change. The diagnosis of Barrett's esophagus is based on the presence of intestinal metaplasia in a biopsy from an endoscopically visualized abnormal columnar epithelium. In this pilot study, acoustic microscopy was used to identify the mucosal structure of 10 distal esophageal biopsies. Sections cut at 5 microm of archival paraffin blocks on glass slides were used for this study. Acoustic microscopy permitted the identification of low- and high-power images of epithelial architecture and cellular detail, including Barrett's epithelium. This modality of visualization has the potential to detect lesions such as Barrett's metaplasia, low- and high-grade dysplasia and early carcinoma. If it can be applied to in vivo endoscopy, acoustic microscopy has the potential to increase the accuracy of the diagnosis of Barrett's esophagus, dysplasia and malignancy by providing a method of accurately directing biopsies at endoscopy. PMID- 11937284 TI - Online artery diameter measurement in ultrasound images using artificial neural networks. AB - An automated online technique is described for measurement of artery diameter in flow-mediated dilation (FMD) ultrasound (US) images, using artificial neural networks to identify and track artery walls. This allows FMD results to be calculated without the inherent delay of current retrospective methods. Two networks were trained to identify artery anterior and posterior walls using over 3200 examples from carotid artery images. Both networks correctly classified approximately 97% of the randomly selected test samples. The technique was verified using a physical model with absolute measurement error of -1.16% +/- 1.04% (mean +/- SD) over the diameter range 2 to 8 mm. Advantages of the technique include: online analysis; wall tracking optimisation before the study proper; measurement of diameter changes over the cardiac cycle; low FMD measurement variance; minimal image degradation; and no unwieldy image store. Measurement of artery diameter changes over the cardiac cycle was explored using simulated image sequences generated with a virtual US scanner. PMID- 11937285 TI - A model based upon pseudo regular spacing of cells combined with the randomisation of the nuclei can explain the significant changes in high-frequency ultrasound signals during apoptosis. AB - Recent ultrasound (US) experiments on packed myeloid leukaemia cells have shown that, at frequencies from 32 to 40 MHz, significant increases of signal amplitude were observed during apoptosis. This paper is an attempt to explain these signal increases based upon a simulation of the backscattered signals from the cells nuclei. The simulation is an expansion of work in which a condensed sample of cells, with fairly regular sizes, could be considered as an imperfect crystal. Thus, destructive interference could occur and this would be observed as a large reduced value of backscattered signals compared with the values obtained from a similar, but random, scattering source. This current paper explores the possibility that simple changes in the nuclei, such as their observed condensation or the small loss of nuclei scatterers from cells, could cause a significant increase in the observed backscattered signals. This model indicates that the greater backscattered signals can be explained by further randomisation of the average positions of the scattering sources in each cell. When these "microechoes" are added together, so that the destructive interference is reduced, a large increase in the signal is predicted. The simplified model strongly suggests that much of observed large increases of the backscattered signals could be simply explained by the randomisation of the position of the condensed nuclei during apoptosis, and the destruction of the nuclei could produce further signal amplitude changes due to disruption of the cloud of backscattered waves. PMID- 11937286 TI - Acoustic radiation force impulse imaging: in vivo demonstration of clinical feasibility. AB - The clinical viability of a method of acoustic remote palpation, capable of imaging local variations in the mechanical properties of soft tissue using acoustic radiation force impulse (ARFI) imaging, is investigated in vivo. In this method, focused ultrasound (US) is used to apply localized radiation force to small volumes of tissue (2 mm(3)) for short durations (less than 1 ms) and the resulting tissue displacements are mapped using ultrasonic correlation-based methods. The tissue displacements are inversely proportional to the stiffness of the tissue and, thus, a stiffer region of tissue exhibits smaller displacements than a more compliant region. Due to the short duration of the force application, this method provides information about the mechanical impulse response of the tissue, which reflects variations in tissue viscoelastic characteristics. In this paper, experimental results are presented demonstrating that displacements on the order of 10 microm can be generated and detected in soft tissues in vivo using a single transducer on a modified diagnostic US scanner. Differences in the magnitude of displacement and the transient response of tissue are correlated with tissue structures in matched B-mode images. The results comprise the first in vivo ARFI images, and support the clinical feasibility of a radiation force based remote palpation imaging system. PMID- 11937287 TI - A Doppler system for dynamic vector velocity maps. AB - The aim of the vector Doppler technique is the quantitative reconstruction of a velocity field independently of the ultrasonic probe axis to flow angle. In particular, vector Doppler is interesting for studying vascular pathologies related to complex blood flows. A problem of vector Doppler is data representation in real-time that should be easy to interpret for the physician. In this work, we present a technique for dynamic display of vector velocity maps and some experimental results obtained in vitro with 2-D vector Doppler on flow phantoms reproducing complex flow conditions. An improvement in the map presentation was obtained by using velocity vector field interpolation. In this work, we considered the problem of spatial sampling for vector Doppler, establishing a relationship between sampling steps and scanning system characteristics. Finally, we developed a novel multimedia solution that uses both interpolated images and sound to discriminate between laminar and turbulent flows. PMID- 11937288 TI - Toward a better quantitative measurement of aortic flow. AB - Ultrasound investigation of aortic blood flow (ABF) still represents a technically challenging task, because of the complex geometry of such a deep artery. In this paper, we present a unique experimental set-up capable of providing detailed information about blood dynamics in the aorta. The set-up is based on an esophageal probe (EP) connected to a multigate Doppler-processing system. The EP, developed for the noninvasive hemodynamic monitoring of ABF in patients under general anesthesia or in the intensive care area, must be inserted at a thoracic depth where the esophagus and the aorta are nearby and parallel. Doppler processing of pulsed wave echoes in the multigate system provides the distribution of all Doppler frequencies detected along the probe beam axis (spectral profile) in real time. The results of this investigation confirm that flow in the aorta is extremely complex, especially at the level of the aortic arch or in nonphysiologic circumstances. In general, the velocity profiles tend to be flat only during the systolic acceleration, but not during the full cardiac cycle. In most cases, they are asymmetrical, including both positive and negative components. In particular, it is shown that an appropriate positioning of the ultrasound transducer and/or the correct integration of different velocities is mandatory to make reliable ABF measurements. PMID- 11937289 TI - Time delays in ultrasound systems can result in fallacious measurements. AB - Even short time delays (less than 30 ms) in cardiac motion pattern may have clinical relevance. These delays can be measured with echocardiography, using techniques such as flow and tissue Doppler and M-mode together with external signals (e.g., ECG and phonocardiography). If one or more of these signals are delayed in relation to the other signals (asynchronous), an incorrect definition of cardiac time intervals can occur, the consequence of which is invalid measurement. To determine if this time delay in signal processing is a problem, we tested three common ultrasound (US) systems using the ECG as the reference signal. We used a digital ECG simulator and a Doppler string phantom to obtain test signals for flow and tissue pulsed Doppler, M-mode, phonocardiography, auxiliary and ECG signals. We found long time delays of up to 90 ms in one system, whereas delays were mostly short in the two other systems. The time delays varied relative to system settings. Consequently, to avoid these errors, precise knowledge of the characteristics of the system being used is essential. PMID- 11937290 TI - Quantitative ultrasound of the hand phalanges and calcaneus revealed skeletal abnormalities due to primary hyperparathyroidism: a case report. AB - Primary hyperparathyroidism (PHPT) is a common disease causing bone loss in elderly patients. We report a case study of a 36-year-old woman with PHPT. Quantitative ultrasound (QUS) assessment of the phalanges and calcaneus revealed significantly lower than normal values for age. This observation was confirmed by measuring bone mineral density in different skeletal sites using dual-energy X ray absorptiometry (DXA). Subsequent parathyroid adenoma surgery normalized calcium metabolism, resulting in a progressive increase of BMD and ultrasound (US) parameters. This report has shown an ability of peripheral QUS examinations (phalanges and calcaneus) in early detection of bone alterations caused by PHPT in a young woman. Skeletal changes after surgery could be evaluated by QUS in a similar manner to that used in DXA. PMID- 11937293 TI - Mechanistic comparisons among base excision repair glycosylases. AB - The mechanisms by which various DNA glycosylases initiate the base excision repair pathways are discussed. Fundamental distinctions are made between "simple glycosylases," that do not form DNA single-strand breaks, and "glycosylases/abasic site lyases," that do form single-strand breaks. Several groupings of BER substrate sites are defined and some interactions between these groupings and glycosylase mechanisms discussed. Two characteristics are proposed to be common among all BER glycosylases: a nucleotide flipping step that serves to expose the scissile glycosyl bond to catalysis, and a glycosylase transition state characterized by substantial tetrahedral character at the base glycosyl atom. PMID- 11937294 TI - Free radicals in alcoholic myopathy: indices of damage and preventive studies. AB - Chronic alcoholic myopathy affects up to two-thirds of all alcohol misusers and is characterized by selective atrophy of Type II (glycolytic, fast-twitch, anaerobic) fibers. In contrast, the Type I fibers (oxidative, slow-twitch, aerobic) are relatively protected. Alcohol increases the concentration of cholesterol hydroperoxides and malondialdehyde-protein adducts, though protein carbonyl concentration levels do not appear to be overtly increased and may actually decrease in some studies. In alcoholics, plasma concentrations of alpha tocopherol may be reduced in myopathic patients. However, alpha-tocopherol supplementation has failed to prevent either the loss of skeletal muscle protein or the reductions in protein synthesis in alcohol-dosed animals. The evidence for increased oxidative stress in alcohol-exposed skeletal muscle is thus inconsistent. Further work into the role of ROS in alcoholic myopathy is clearly warranted. PMID- 11937296 TI - Superoxide limits cyclosporine-A-induced formation of peroxynitrite in endothelial cells(2). AB - Peroxynitrite (ONOO(-)) is a potent oxidant formed by the nonenzymatic reaction between superoxide anion (O(2)(*-)) and nitric oxide (NO*) in a one-to-one stoichiometry. Accumulated evidence suggests that endothelial dysfunction coincides with an enhanced NO* synthase expression and O(2)(*-) production, facilitating ONOO(-) formation. In vivo, formation of ONOO(-) has been associated with atherosclerosis and vascular aging. The immunosuppressor Cyclosporine A (CsA) has been associated to human endothelial dysfunction and accelerated atherosclerosis. We have previously shown that CsA induced a transcriptionally mediated increase of the eNOS gene expression and that CsA induced the formation of nitric oxide, O(2)(*-), and ONOO(-) in vascular endothelial cells. In this work, we evaluate the CsA-induced relative amounts of formation of O(2)(*-) and NO*, providing data consistent with a role of O(2)(*-), and not NO*, as the limiting factor in the CsA-dependent intracellular formation of ONOO(-) in vascular endothelial cells. Furthermore, when endothelial cells were treated with CsA in a situation of increased generation of superoxide such as that provided by high glucose levels, a further increase in the formation of peroxynitrite was detected. The temporal availability of O(2)(*-) for peroxynitrite formation may thus become critical in the pathophysiological scenarios where reactive nitrogen intermediates are operative. PMID- 11937295 TI - Reactive oxygen species accelerate production of vascular endothelial growth factor by advanced glycation end products in RAW264.7 mouse macrophages. AB - Advanced glycation end products (AGEs) are believed to play an important role in the development of angiopathy in diabetes mellitus. Previous reports suggested a correlation between accumulation of AGEs and production of vascular endothelial growth factor (VEGF) in human diabetic retina. However, the mechanisms involved were not revealed. In this study, we investigated the transcriptional regulation of the expression of vascular endothelial growth factor (VEGF) by AGEs, and possible involvement of reactive oxygen species (ROS) in the induction. We employed an AGE of bovine serum albumin (BSA) prepared by an incubation of BSA with D-glucose for 40 weeks and N(epsilon)-(carboxymethyl)lysine (CML), a major AGE. The expression of VEGF was induced by CML-BSA in RAW264.7 mouse macrophage like cells. CML-BSA stimulated the DNA-binding activity of activator protein-1 (AP-1). Promoter assay showed that the induction of VEGF was dependent on AP-1. The activity of Ras/Raf-1/MEK/ERK1/2 was involved in the CML-BSA-stimulated signaling pathways to activate the AP-1 transcription with a peak at 1 h. AGE-BSA also induced VEGF mediated by AP-1, however, there was a difference of effect between AGE-BSA and CML-BSA in the activation of AP-1. AGE-BSA-stimulated AP-1 activity showed a peak at 5 h, which paralleled the formation of ROS. Reduction of AGE-BSA with NaBH(4) or addition of vitamin E attenuated the AGE-BSA stimulated signaling pathways leading to the same pattern as for CML-BSA stimulated signals. These results suggest an important role for AGEs in stimulation of the development of angiogenesis observed in diabetic complications, and that ROS accelerates the AGE-stimulated VEGF expression. PMID- 11937297 TI - Polynitroxyl-albumin (PNA) enhances myocardial infarction therapeutic effect of tempol in rat hearts subjected to regional ischemia-reperfusion. AB - Tempol (4-hydroxy-2,2,6,6-tetramethylpiperidine-N-oxyl, TPL), a low molecular weight stable nitroxyl radical (nitroxide), has been demonstrated in many in vitro and in vivo models to have protective effects against oxidative stress. The beneficial effect of TPL, however, is limited because of its short life-time in tissues. We have previously shown that polynitroxylated macromolecules such as polynitroxyl-human serum albumin (PNA) enable maintaining a sustained concentration of TPL for longer periods in tissues. PNA itself has previously been shown to inhibit ischemia-reperfusion (I/R) injury in the gut and to potentiate the activity of TPL. The aim of the present study was (i) to select an optimum formulation of PNA + TPL for therapeutic applications using in vivo EPR spectroscopy and (ii) to evaluate the efficacy of the PNA + TPL formulation in preventing I/R injury to heart, in an in vivo rat model. Rats were subjected to 45 min occlusion of the left anterior descending (LAD) coronary artery followed by 120 min reperfusion. PNA (100 mg/ml) + TPL (10 mg/ml), human serum albumin (HSA, 100 mg/ml) + TPL (10 mg/ml), or saline were injected 5 min before ischemia (3 ml/kg BW, i.v.) and 5 min before reperfusion (3 ml/kg BW, i.v.), followed by a 4 ml/kg BW infusion over 2 h reperfusion. Myocardial risk and infarct regions were then estimated. The results showed that the infarct volume, expressed as a percentage of the risk region, in the group treated with PNA + TPL was 39.7 +/- 3.1%, which was significantly smaller than for the saline (51.3 +/- 3.5%) or HSA + TPL (48.4 +/- 1.4%) groups. The results demonstrate that the PNA + TPL combination is very effective in reducing myocardial ischemia-reperfusion injury. PMID- 11937298 TI - Aggregation of eumelanin mitigates photogeneration of reactive oxygen species. AB - Melanins protect tissue by absorption and rapid nonradiative, nonreactive dissipation of ultraviolet (UV) light. However, melanins also produce reactive oxygen species (ROS) upon UV illumination. A chemical understanding of this dichotomy of photoprotection and phototoxicity has not been established. Herein this issue is examined by studying the UV-B induced oxidation and reduction of cytochrome c by ROS generated by different aggregation states of eumelanin. The quantum yield for superoxide anion by unaggregated oligomers is 7.4 x 10(-3), an order of magnitude greater than that characteristic of the bulk pigment. The quantum efficiency of hydrogen peroxide production by oligomers is 5.7 x 10(-3), and its production is attributed to reaction between superoxide anion and hydroquinone groups on eumelanin oligomers. Aggregation of oligomers results in a reduction of these quantum yields, having a significantly greater effect on the efficiency of hydrogen peroxide production. This effect is attributed to the decrease in surface concentration of hydroquinone sites upon aggregation. The effect of aggregation on the photogeneration of ROS serves to provide a foundation for the understanding of the dichotomy of photoprotective and phototoxic properties of melanin. PMID- 11937299 TI - Critical role of an endogenous gastric peroxidase in controlling oxidative damage in H. pylori-mediated and nonmediated gastric ulcer. AB - The objective of the present study is to delineate the mechanism of oxidative damage in human gastric ulcerated mucosa despite the presence of some antioxidant enzymes. We report for the first time the critical role of an endogenous peroxidase, a major H(2)O(2) metabolizing enzyme, in controlling oxidative damage in gastric mucosa. Human gastric mucosa contains a highly active peroxidase in addition to the myeloperoxidase contributed by neutrophil. In both non Helicobacter pylori (H. pylori)- and H. pylori-mediated gastric ulcer, when myeloperoxidase level increases due to neutrophil accumulation, gastric peroxidase (GPO) level decreases significantly. Moreover, gastric ulcer is associated with oxidative damage of the mucosa as evidenced by significant increase in lipid peroxidation, protein oxidation, and thiol depletion indicating accumulation of reactive oxygen metabolites (ROM). Mucosal total superoxide dismutase (Mn and Cu-Zn SOD) level also decreases significantly leading to increased accumulation of O(2)(*-). To investigate the plausible ROM-mediated inactivation of the GPO during ulceration, the enzyme was partially purified from the mucosa. When exposed to an in vitro ROM generating system, using Cu(2+), ascorbate, and H(2)O(2,) the enzyme gets inactivated, which is dependent on Cu(2+), ascorbate, or H(2)O(2). Insensitivity to SOD excludes inactivation by O(2)(*-). However, complete protection by catalase indicates that H(2)O(2) is essential for inactivation. Sensitivity to EDTA and hydroxyl radical *OH) scavengers indicates that GPO is inactivated most probably by *OH generated from H(2)O(2). We propose that GPO is inactivated in vivo by ROM generated by activated neutrophil. This leads to further accumulation of endogenous H(2)O(2) to cause more oxidative damage to aggravate the ulcer. PMID- 11937300 TI - Genetic control of neutrophil superoxide production in diabetes-resistant ALR/Lt mice. AB - The neutrophil oxidative burst reaction differentiates ALR/Lt mice, known for an unusual systemic elevation of antioxidant defenses, from ALS/Lt mice, a related strain known for reduced ability to withstand oxidative stress. Neutrophils from marrow of ALS mice produced a normal neutrophil oxidative burst following phorbol ester stimulation. In contrast, ALR mice exhibited a markedly suppressed superoxide burst. F1 progeny from reciprocal outcrosses between ALR and ALS mice exhibited an intermediate burst level, higher than ALR but significantly lower than ALS. To elucidate the genetic basis for this strain difference, F1 mice were backcrossed to ALS mice, and marrow neutrophils isolated from the progeny were phenotyped for oxidative burst capacity. A genome-wide sweep using polymorphic markers distinguishing the two parental strains was performed to map the trait. A 1:1 phenotypic distribution was observed, and a locus (Suppressor of superoxide production, Susp) controlling this phenotype was mapped to Chromosome 3 near D3Mit241 at 33.1 cM. This locus probably represents an important regulatory element in the overall ALR strain resistance to oxidative stress, since diminished ability to mount a neutrophil burst in backcross segregants correlated with elevated hepatic superoxide dismutase 1 (SOD1) activity, an ALR strain characteristic. PMID- 11937301 TI - Heat shock inactivates cellular antioxidant defenses against hydrogen peroxide: protection by glucose. AB - Hyperthermia is used in cancer treatment and potentiates the cytotoxicity of radiation and certain chemotherapy drugs. The mechanism(s) of heat killing and those involved in heat potentiation of cytotoxic modalities are not understood. This study examines whether heat shock causes a redox imbalance, leading to oxidative changes in Chinese hamster ovary cells. Decreases in the GSH/GSSG ratio reflected an oxidative imbalance in heated (42 degrees C) and in H(2)O(2) challenged cells. Glucose provided protection against these changes. Glucose also protected cells against cytotoxicity of H(2)O(2) and/or hyperthermia (42 to 43 degrees C). Glucose appears to protect cells against H(2)O(2) and heat shock by providing NADPH through its metabolism via the pentose phosphate cycle (PC). When cells were deprived of glucose, there was a marked decrease in the GSH/GSSG ratio and in NADPH levels, indicating a severe redox imbalance. Glucose deprivation caused cell death, which was consistent with increased accumulation of H(2)O(2), since three distinct H(2)O(2)-detoxifying systems (N-acetyl-L-cysteine, sodium pyruvate, and catalase) rescued cells against cytotoxicity. Nontoxic levels of H(2)O(2) stimulated a corresponding increase in both PC activity and NADPH levels. NADPH levels and basal activity of the PC increased at 42 degrees C. However, the oxidant-stimulated increases in PC activity and NADPH levels were lost in heated cells. Therefore, heat shock inactivates an important cellular defense mechanism against oxidants. These findings suggest that heat shock may enhance the cytotoxicity of oxidants by inhibiting increases in PC activity following oxidative stress. These data are potentially relevant to understanding the potentiation of cytotoxicity of radiation and oxidant-generating drugs by heat shock, used in combined modality cancer treatment. PMID- 11937302 TI - Biosynthetic incorporation of oxidized amino acids into proteins and their cellular proteolysis. AB - We demonstrate that oxidized amino acids can be incorporated into proteins by protein synthesis. The level of incorporation into protein was dependent on the concentration of oxidized amino acid supplied to the cells. At low levels of incorporation, the oxidized amino acids examined increased the degradation rate of the cell proteins. Degradation of certain proteins containing high levels of DOPA (but not ortho or meta tyrosine) was decreased to below the basal degradation rates suggesting that DOPA may contribute to proteins becoming resistant to proteolysis. Changes in the degradation rates of the oxidized amino acid-containing proteins was shown to have no impact on the degradation rates of native proteins, indicating that the activity of the degradative machinery was not affected. We demonstrate that oxidized proteins are selectively degraded by the proteasomes and provide evidence to suggest that the proteasomes and the endosomal-lysosomal systems may act in sequence as well as in parallel. The incorporation approach, unlike cell studies in which an exogenous oxidant is used, allows the degradation rates of the oxidatively modified proteins to be selectively measured, offering a greater sensitivity as well as greatly reducing toxicity to the cell and avoiding oxidative modification of other cell components. PMID- 11937303 TI - Neuropeptide Y-Y1 receptor modulates nitric oxide level during stroke in the rat. AB - In a rat endovascular middle cerebral artery occlusion (MCAO) stroke model, we previously showed that intracerebroventricular (ICV) injection of neuropeptide Y (NPY) or an Y1 receptor agonist, [Leu(31),Pro(34)]-NPY, increased the infarct volume, that an Y1 receptor antagonist, BIBP3226, reduced the infarct volume, and that an Y2 receptor agonist, NPY3-36, had no effect. In this study, we used electron paramagnetic resonance (EPR) spectroscopy to measure nitric oxide (NO) and examined how ICV administration of NPY or its receptor analogs would modulate the brain NO level between the bregma levels +2 and -4 mm during MCAO, since excessive NO mediates ischemic damage. The relative brain NO concentration was increased to 131.94 +/- 7.99% (mean +/- SEM; n = 8) at 15 min of MCAO. NPY treatment further increased the relative brain NO concentration to 250.94 +/- 50.48% (n = 8), whereas BIBP3226 significantly reduced the brain NO concentration to 69.63 +/- 8.84% (n = 8). [Leu(31),Pro(34)]-NPY (137.61 +/- 14.54%; n = 7) or NPY3-36 (129.23 +/- 21.77%; n = 8) did not affect the brain NO concentration at 15 min of MCAO. Our results suggest that the NPY-Y1 receptor activation mediates ischemic injury via NO overproduction and that inhibition of the Y1 receptor may confer protection via suppression of excessive NO production during ischemia. PMID- 11937305 TI - Comment on "The implementation of guidelines and computerised forms improves the completeness of cancer pathology reporting. The CROPS project: a randomised controlled trial in pathology" by Branston and colleagues. PMID- 11937306 TI - Decision-making in early breast cancer: guidelines and decision tools. AB - The meta-analysis of trials of adjuvant systemic therapy for early breast cancer provides robust information on the impact of both cytotoxic chemotherapy and tamoxifen on relapse-free and overall survival to 15 years from diagnosis. These data are described in terms of relative risk reduction and are not meant to be viewed as a prescription for therapy. To translate relative risk reductions into absolute benefits for the individual patient and then trade off the gains against the long-term and short-term side-effects and toxicities is a highly complex process for the clinician, and current guidelines are formatted in such a way that they fail to use current information in a way that allows a quantitative assessment of the benefits and risks of adjuvant therapy. This review article explores current guidelines and describes some aids that may be used to help inform women about their treatment options for early breast cancer. PMID- 11937307 TI - Angiogenesis and anti-angiogenesis in neuroblastoma. AB - Angiogenesis is a biological process by which new capillaries are formed from pre existing vessels. It occurs in physiological and pathological conditions, such as tumours, where a specific critical turning point is the transition from the avascular to the vascular phase. Tumour angiogenesis depends mainly on the release by neoplastic cells of growth factors specific for endothelial cells that able to stimulate the growth of the host's blood vessels. This review summarises the literature concerning the relationship between angiogenesis and progression in human neuroblastoma, the most common extracranial solid tumour of infancy and childhood. It is becoming increasingly evident that agents which interfere with blood vessel formation also block tumour progression. Accordingly, anti angiogenic tumour therapy has gained much interest in preclinical and clinical assessments. The recent applications of anti-angiogenic agents which interfere or block neuroblastoma progression are reviewed. PMID- 11937308 TI - Cytokeratins 20 and 7 as biomarkers: usefulness in discriminating primary from metastatic adenocarcinoma. AB - Metastatic adenocarcinoma from an unknown primary site is a common clinical problem. The use of cytokeratins 20 (CK20) and 7 (CK7) was proposed to identify the primary sites in this situation. In this review, the results of 29 studies were summarised and the difficulties of data comparison described. Most tumours retained the CK20 phenotype during metastasis, but lung, non-mucinous ovarian, and gastric adenocarcinomas showed statistically significant differences in CK20 expression in the reported primary and metastatic cases. Ductal breast carcinomas, lung and non-mucinous ovarian adenocarcinomas showed significant differences in CK7 expression when primary and metastatic tumours were compared. CK20 positivity alone indicates metastatic spread of adenocarcinoma in several organs. CK7 negativity is consistent with metastases of adenocarcinomas in the lungs, ovaries, liver or serous membranes. CK20/7 phenotyping of adenocarcinomas is a useful diagnostic tool if based on algorithmic and probabilistic approaches and a detailed database. PMID- 11937309 TI - The implementation of guidelines and computerised forms improves the completeness of cancer pathology reporting. The CROPS project: a randomised controlled trial in pathology. AB - The aim of this study was to determine whether reporting guidelines and computerised form-based reports improve the completeness of histopathological cancer data available for patient management and population cancer registration and to evaluate the acceptability of the intervention. The study was a randomised controlled trial with a split unit design and stratified cluster randomisation. All 16 hospital pathology laboratories in Wales were randomly allocated to report either breast or colorectal resection specimens by computerised form or conventional free text. 1044 reports were analysed in the study arm, 998 in the control arm. Use of pre-defined forms led to a 28.4% (95% confidence interval (CI): 15.7-41.2%) increase in complete reporting of a minimum dataset required for cancer registration and a 24.5% (95% CI: 11.0-38.0%) increase in complete reporting of minimum data required for patient management. Form-based reporting was acceptable to pathologists and preferred by clinicians. In conclusion, guidelines and computerised forms significantly improve the quality of histopathology reporting. PMID- 11937310 TI - An EORTC-IDBBC phase I study of gemcitabine and continuous infusion 5 fluorouracil in patients with metastatic breast cancer resistant to anthracyclines or pre-treated with both anthracyclines and taxanes. AB - The aim of this study was to determine the maximum tolerated dose (MTD), dose limiting toxicities (DLT), and potential activity of combined gemcitabine and continuous infusion 5-fluorouracil (5-FU) in metastatic breast cancer (MBC) patients that are resistant to anthracyclines or have been pretreated with both anthracyclines and taxanes. 15 patients with MBC were studied at three European Organization for Research and Treatment of Cancer centres. 13 patients had received both anthracylines and taxanes. Gemcitabine was given intravenously (i.v.) on days 1 and 8, and 5-FU as a continuous i.v. infusion on days 1 through to 14, both drugs given in a 21-day schedule at four different dose levels. Both were given at doses commonly used for the single agents for the last dose level (dose level 4). One of 6 patients at level 4 (gemcitabine 1200 mg/m2 and 5-FU 250 mg/m2/day) had a DLT, a grade 3 stomatitis and skin toxicity. One DLT, a grade 3 transaminase rise and thrombosis, occurred in a patient at level 2 (gemcitabine 1000 mg/m2 and 5-FU 200 mg/m2/day). Thus, the MTD was not reached. One partial response and four disease stabilisations were observed. Only 1 patient withdrew from the treatment due to toxicity. The MTD was not reached in the phase I study. The combination of gemcitabine and 5-FU is well tolerated at doses up to 1200 mg/m2 given on days 1 and 8 and 250 mg/m2/day given on days 1 through to 14, respectively, every 21 days. The clinical benefit rate (responses plus no change of at least 6 months) was 33% with one partial response, suggesting that MBC patients with prior anthracycline and taxane therapy may derive significant benefit from this combination with minimal toxicity. PMID- 11937311 TI - A phase II EORTC study of temozolomide in patients with malignant pleural mesothelioma. AB - The aim of this study was to investigate the anti-tumour activity of temozolomide in patients with malignant pleural mesothelioma. 27 chemotherapy-naive patients with histologically-proven malignant mesothelioma were treated with temozolomide 200 mg/m2/day, given orally on days 1-5 of each 28-day cycle. Therapy continued up to 10 cycles unless disease progression or excessive toxicity mandated discontinuation. Toxicity, symptom improvement and pain intensity were regularly assessed. With a median relative dose intensity of 97%, toxicity was moderate with grade 3 or more nausea, vomiting, thrombocytopenia, leucocytopenia, neutropenia, febrile leucocytopenia, arthralgia, infection and fever with infection occurring in 13, 13, 10, 3, 7 and 3% of patients for the remaining events, respectively. Overall, 1 objective response was observed (response rate 4%, 95% Confidence Interval (CI): 0.1-19). Median survival was 8.2 months. Symptom assessment showed no improvement and an increase of pain was observed during the study. Thus, oral temozolomide is an inactive agent in malignant mesothelioma. PMID- 11937312 TI - The role of dynamic imaging in sentinel lymph node biopsy in breast cancer. AB - The aim of this study was to evaluate the role of dynamic imaging in sentinel lymph node (SLN) biopsy in breast cancer. Patients with T1/T2, N0 invasive breast cancer underwent SLN localisation using intra-dermal injection of 15 MBq of 99mTc nanocolloid. Gamma camera anterior-oblique dynamic imaging commenced simultaneously with tracer administration for 45 min, and was followed by anterior and lateral static imaging. Dynamic imaging data was reformatted into image files of different time-frames. Patterns of uptake were analysed using the sequences of dynamic frames and time-activity curve (TAC). SLN localisation was successful in 70/73 studies (96%) in 72 patients. Imaging information was present within the first 15 min of dynamic imaging in 67/70 studies (96%). Critical analysis of dynamic data helped to differentiate true SLN from secondary echelon nodes in eight studies and transient foci of radioactivity in six studies. In 17 studies, SLN contained metastatic disease. The detection of SLN metastasis was independent from the use of dynamic imaging. Dynamic imaging improves the interpretation of preoperative SLN imaging for breast cancer, but does not contribute significantly to the successful detection of SLN. Hence, preoperative dynamic imaging is not necessary in SLN biopsy for breast cancer. PMID- 11937313 TI - Symptomatology of cancer patients in palliative care: content validation of self assessment questionnaires against medical records. AB - To elucidate which symptoms or problems to measure when evaluating palliative care, we assessed the content validity of selected patient self-assessment questionnaires used to evaluate palliative care: the European Organization for Research and Treatment of Cancer-Quality of Life-Core 30 (EORTC QLQ-C30), the Edmonton Symptom Assessment System (ESAS), the Palliative Care Outcome Scale (POS), the McGill Quality of Life Questionnaire (MQOL) and the Memorial Symptom Assessment Scale (MSAS). The content of the questionnaires was compared against the symptoms and problems noted in the medical records of 171 consecutive cancer patients on their first admission to a department of palliative medicine. From the records, 63 different symptoms were listed. Two questionnaires covered almost all of the prevalent symptoms/problems: the EORTC QLQ-C30 covered 10 and the MSAS 11 of the 12 most frequent problems. Researchers selecting instruments for evaluating palliative care may use the present study and other reviews to examine to what degree a given selection of instruments cover the symptoms/problems targeted by palliative care physicians. PMID- 11937314 TI - A population-based study of intensive multi-agent chemotherapy with or without autotransplant for the highest risk Hodgkin's disease patients identified by the Scotland and Newcastle Lymphoma Group (SNLG) prognostic index. A Scotland and Newcastle Lymphoma Group study (SNLG HD III). AB - The aim of the study was to identify all patients with poor risk Hodgkin's disease (HD) using a numerical prognostic index in a defined population and to recruit them into a trial of intensive chemotherapy prednisolone, vinblastine, doxorubicin, chlorambucil, etoposide, bleomycin, vincristine, procarbazine (PVACE BOP)x3+autotransplant (Arm A) versus PVACE-BOPx5 (Arm B) in first remission. In 10 years, the Scotland and Newcastle Lymphoma Group (SNLG) registered 930 patients with HD of whom 178 (19%) were identified as 'poor risk' by the SNLG index and were aged 16-59 years. 126/178 (71%) entered the study. Of the 120 confirmed poor risk HD cases, all completed PVACE-BOPx3 with a 93% Complete Response/unconfirmed Complete Response (CR/CRu) rate. Only 65/107 in CR accepted the randomisation. With a median follow-up of 6 years, both arms of the trial have a similar time to treatment failure (TTF) (Arm A 79%+/-11 versus 85%+/-7 Arm B, P=0.35). Advanced stage 'good risk' patients not included in the trial receiving standard therapy with CLVPP or ABVD had a 75% 5-year survival. The study demonstrates that PVACE-BOP therapy in the poorest risk group (58% had an IPI>/=3) produces excellent CR rates (93%) and overall survival with minimal toxicity, and that the substitution of autotransplant in first CR does not improve outcome. The use of the objective SNLG index accurately helped in the selection of the poorest risk group in this population study. The placing of a randomised control trial within the context of a population-based study of HD enhances the validity of the outcome. PMID- 11937315 TI - Clinical phase II study and pharmacological evaluation of rubitecan in non pretreated patients with metastatic colorectal cancer-significant effect of food intake on the bioavailability of the oral camptothecin analogue. AB - A randomised, open label phase II study was performed in patients with advanced colorectal cancer to evaluate the safety, toxicity and antineoplastic activity of the topoisomerase I-inhibitor rubitecan. A cross-over design was chosen to determine the intrapatient variation of the bioavailability and pharmacokinetics of the anticancer agent depending on the timing of food intake in relation to the oral drug administration. Patients with previously untreated metastatic disease received two single oral doses of rubitecan 1.5 mg/m2 for assessment of the pharmacokinetics. They were randomised to have the first administration either after an overnight fasting period or immediately after a high calorie breakfast, and crossed over to the alternative schedule after a one-week washout period. After completion of the pharmacokinetic sampling, treatment continued with rubitecan given orally at a dose of 1.5 mg/m2/day, to be increased up to 2.0 mg/m2/day, under fasting conditions for 5 consecutive days per week until disease progression. 19 patients entered the trial after informed consent was obtained. A total number of 35 treatment cycles (median 2, range 1-4) were administered. All patients were evaluable for safety. The toxicity profile of rubitecan was generally mild to moderate, with sporadic cases of grade 4 toxicities (Common Toxicity Criteria (CTC) version 2.0) diarrhoea, leucopenia and neutropenia. None of 15 evaluable patients achieved an objective response. The majority had early disease progression. 14 patients were evaluable for pharmacokinetic analysis. The bioavailability of rubitecan was found to be strongly dependent on the timing of food intake with a fasted-to-fed ratio for C(max) of 1.98 (two-tailed P<0.001; ANOVA), T(max) 0.49 (P<0.001), AUC(0-8 h) 2.52 (P<0.001) and AUC(0-24 h) 1.64 (P=0.003). Rubitecan is well tolerated, but clinically inactive in colorectal cancer at the currently recommended dose and schedule. The bioavailability is strongly dependent on the timing of food intake in relation to the oral administration of the drug. The topoisomerase I-inhibitor should be administered under fasting conditions to achieve adequate drug exposure in future prospective trials in other tumour types. PMID- 11937317 TI - Sun exposure and sun protection in young European children: an EORTC multicentric study. AB - Most European children experience exposure to the sun during the summer holidays. The aim of this study was to examine the behaviour of European children when in the sun during their holidays. In 1995-1997, a total of 631 young children were recruited during a multicentric study in Belgium, Germany, France and Italy. For each holiday period from birth, parents gave detailed information on sun exposure and child behaviour. Predictors and trends over time of sun protection were investigated. Forty percent of children were exposed to sunlight in the first and 86% in the sixth year of life. At the same time, the number of children who experienced sunburns rose from 1 to 23%. In the whole period of 6 years, only 8% of children always wore trousers and shirt when in the sun, while 25% children always used a sunscreen. The proportion of sun-exposed children who used sunscreen was stable with age (approximately 50%), while those who always wore trousers and shirts dropped from 46% (1st year) to 19% (6th year). Multinomial logistic regression showed that sunscreen use, but not the wearing of clothes was associated with sun-sensitivity. In summary, sun exposure increases steadily, while sun protection decreases in the first 6 years of life in our cohort of children. In this cohort, use of a sunscreen was much more frequent than the wearing of clothes and a reduction in sun exposure. PMID- 11937316 TI - Carboplatin before and during radiation therapy for the treatment of malignant brain stem tumours: a study by the Societe Francaise d'Oncologie Pediatrique. AB - Childhood malignant brain stem tumours have a very poor prognosis with a median survival of 9 months despite radiotherapy. No chemotherapy has improved survival. However, carboplatin has been reported to have activity in glial tumours as well as antitumour synergy with radiation. Our aims were to test the response rate of these tumours to carboplatin alone and to evaluate the efficacy on survival of carboplatin alone followed by concurrent carboplatin and radiotherapy. Patients younger than 16 years with typical clinical and radiological presentation of infiltrating brain stem tumour, as well as histologically-documented cases in the atypical forms, were eligible. Two courses of carboplatin (1050 mg/m2 over 3 days) were administered initially. This treatment was followed by a chemoradiotherapy phase including five weekly carboplatin courses (200 mg/m2) and conventional radiotherapy. 38 eligible patients were included. No tumour response was observed after the initial phase. This schedule of first-line carboplatin followed by concurrent carboplatin and radiotherapy did not improve survival. PMID- 11937318 TI - Women who participate in spontaneous screening are not at higher risk for cervical cancer than women who attend programme screening. AB - Up to 1995, programme screening for cervical cancer in The Netherlands was targeted at women between 35 and 54 years of age at 3-yearly intervals. Spontaneous screening in addition to programme screening was common practice. Our aim was to compare the underlying risk for cervical neoplasia for women involved in both types of screening. From the national pathological database, we retrieved all primary smears (n=693318) taken in 1994 in The Netherlands. Among the smears registered for screening purposes (39%), 79% was taken within the mass screening programme and 21% was taken for spontaneous screening. The underlying risk was studied from the detection rates of histologically confirmed severe dysplasia or worse, using a multivariate loglinear model, including age and screening history. The detection rate of at least severe dysplasia, adjusted for age and screening history, was equal for women who had a spontaneous smear and for those who had a programme smear (odds ratio (OR): 0.97; 95% Confidence Interval (CI): 0.84-1.14). In our data, women participating in spontaneous screening were not at a higher risk for cervical cancer than women who used programme screening. Therefore, all asymptomatic women in the Netherlands should follow the general guidelines for age-range and screening-interval. PMID- 11937319 TI - Alterations in p53 and pRb pathways and their prognostic significance in oesophageal cancer. AB - The pRb (p16-pRb-cyclin D1) and p53 (p53-MDM2-p21) pathways play a critical role in tumorigenesis. To evaluate which of these cell cycle regulatory proteins are related to patients' prognosis, a comprehensive analysis of alterations in these components was carried out in 100 ESCCs (oesophageal squamous cell carcinoma) using immunohistochemistry and correlated with clinicopathological parameters by univariate analysis. Overexpression of p53, MDM2 and cyclin D1 proteins was observed in 73, 42 and 67% of the cases, respectively, while loss of expression of p21, p16 and pRb was observed in 36, 45 and 75% of the cases, respectively. Multiple logistic regression analysis revealed that loss of p16 immunoreactivity was a significant risk factor for tumour stage (pT) (Odds Ratio (OR)=3.3), whereas the loss of pRb was a significant risk factor for nodal metastasis (pN) (OR=8.8). MDM2 overexpression emerged as the most significant risk factor for distant organ metastasis (pM) (OR=4.6). Of the ESCC patients who underwent oesophagectomy, 50 cases were followed-up for a maximum period of 44 months and median of 16 months. Survival analysis revealed that Cyclin D1 overexpression is an adverse prognosticator for disease-free survival, as well as overall survival, and tumour stage (pT) is an adverse prognosticator for disease-free survival. In conclusion, these data support a model of oesophageal cancer pathogenesis in which both the pRb and p53 pathways are inactivated and suggests an in-depth evaluation of the clinical utility of these putative markers is warranted. PMID- 11937320 TI - The importance of drug scheduling and recovery phases in determining drug activity. Improving etoposide efficacy in BCR-ABL-positive CML cells. AB - K562 leukaemic cells are known to be less sensitive to etoposide than other cell lines, despite having similar topo II mRNA levels and cleavable complex formation. We have investigated the effect of etoposide schedule on cell cycle distribution, apoptosis and p21(waf1) and cdk1(p34) status in two bcr-abl positive chronic myeloid leukaemia (CML) cell lines (K562 and KU812) and two small cell lung cancer (SCLC) cell lines (H69 and GLC4). During a continuous 5 day exposure, the SCLC cell lines showed a time and concentration-dependent loss of cell viability, with an initial block in the G2/M phase of the cell cycle followed by apoptosis. In contrast, the two CML cell lines showed no significant apoptosis or loss of viability after a similar block in G2/M. However, when K562 or KU812 cells were placed in drug-free medium following a 3-day drug exposure there was marked, concentration-dependent apoptosis (% apoptosis after release at 1 microM etoposide in K562, 10% at 24 h, 30% at 48 h). Our data also show that p21(waf1) does not increase after etoposide treatment in either H69 or GLC4 (both with mutated-p53). Although K562 and KU812 cells are null-p53, the arrest in G2/M during drug exposure was associated with increased p21(waf1) and a decrease in cdk1 (both P<0.001 compared with controls). Upon release of these cells from drug medium, p21(waf1) gradually returned to control levels, which was associated with an easing of the block at G2/M and an induction of apoptosis. This study highlights the importance of cell cycle regulatory proteins in drug sensitivity and resistance, and suggests that in cells such as K562 and KU812, a pulsed schedule may be more active than a single prolonged exposure. PMID- 11937321 TI - Comments on: Extended-term effects of head and neck irradiation in a rodent. Eur J Cancer 2001, 37, 1938-1945. PMID- 11937323 TI - Novel 4,5-diaryl-3-hydroxy-2(5H)-furanones as anti-oxidants and anti-inflammatory agents. AB - In order to study the effect of phenol moieties on biological activities of ascorbic acid derivatives, we synthesized 13 novel 4,5-diaryl-3-hydroxy-2(5H) furanones 5a-m with various substitution patterns. Compound 5 g bearing a 2,3 dihydroxy phenyl ring on the 5-position of the heterocycle appeared to be the most powerful anti-oxidant furanone with reducing activity against DPPH (IC(50)=10.3 microM), superoxide anion quenching capacity (IC(50)=0.187 mM) and lipid peroxidation inhibitory effect (IC(50)=0.129 mM). To ascertain determinant molecular features for anti-oxidant activities, structure-activity relationships were studied. Lipophilicity and molecular parameters related to electron distribution and structure (difference in heats of formation between the compound and its radical or its cation radical, energy of the highest occupied molecular orbital, HOMO) were found to correlate with the anti-oxidant action of compounds 5 in the different tests used. Oxygen-derived free radicals are known to contribute to inflammatory disorders; therefore we have investigated effects of compounds 5 in two models of inflammation: phorbol ester-induced ear edema in mice (TPA-test) and carrageenan-induced paw edema in rat. At 100 mg/kg ip in the TPA-test, the anti-inflammatory activity of compounds 5 was potent compared with that of indomethacin and ketorolac and all the results suggested a cyclooxygenase inhibition in the emergence of such properties. The combined pharmacological actions of compounds 5 associated with a favorable therapeutic index prompt with interesting perspectives for their use in heart and brain disorders as well as in inflammatory diseases. PMID- 11937324 TI - New synthetic siderophores and their beta-lactam conjugates based on diamino acids and dipeptides. AB - Linking of siderophores to antibiotics improves the penetration and therefore increases the antibacterial activity of the antibiotics. We synthesized the acylated catecholates and hydroxamates as siderophore components for antibiotic conjugates to reduce side effects of unprotected catecholate and hydroxamate moieties. In this paper, we report on bis- and tris-catecholates and mixed catecholate hydroxamates based on diamino acids or dipeptides. These compounds were active as siderophores in a growth promotion assay under iron limitation. Most of the conjugates with beta-lactams showed high in vitro activity against Gram-negative bacteria especially Pseudomonas aeruginosa, Escherichia coli, Klebsiella pneumoniae, Serratia marcescens and Stenotrophomonas maltophilia. The compounds with enhanced antibacterial activity use active iron uptake routes to penetrate the bacterial outer membrane barrier, demonstrated by assays with mutants deficient in components of the iron transport system. Correlation between chemical structure and biological activity was studied. PMID- 11937325 TI - Phenylpiperazinylmethylindolecarboxylates and derivatives as selective D(4) ligands. AB - Novel phenylpiperazinylmethylindolecarboxylates were synthesized for evaluation as potential D(4)-ligands. Test compounds showed high affinity for the human dopamine D(4) receptor and great selectivity over the other receptor subtypes. Intrinsic effects of indole derivatives, which indicated most promising binding properties, were investigated in a mitogenesis assay. PMID- 11937326 TI - Anti-Candida albicans properties of novel benzoxazine analogues. AB - We previously reported that azole 1,4-benzothiazine derivatives have appreciable anti-Candida activity. In this study, we synthesized 1,4-benzoxazine analogues and examined their possible antifungal activity to further analyze the structure activity relationships. Results of in vitro and in vivo experiments showed that 1,4-benzoxazine analogues show appreciable antifungal activity. In particular, they have significant capability to cure mice systemically infected with a lethal challenge of Candida albicans, as indicated by increased survival time paralleling reduction of colony forming units. Moreover, 1,4-benzoxazine derivatives also showed immunomodulating activity, as indicated by a significant increase of interleukin-12 and interferon-gamma production by splenocytes and reinforcement of a T helper type 1 protective immune response to C. albicans. In conclusion, the results demonstrate that replacement of sulfur by oxygen may improve immune response against C. albicans infection. PMID- 11937327 TI - Synthesis of a novel quinoline derivative, 2-(2-methylquinolin-4-ylamino)-N phenylacetamide--a potential antileishmanial agent. AB - Some novel quinoline derivatives were prepared and tested for antileishmanial activity. 2-(2-Methylquinolin-4-ylamino)-N-phenylacetamide (2) was found to be significantly more active than the standard antileishmanial drug sodium antimony gluconate (SAG) in reducing the parasite load both in the spleen and liver at a much lower concentration in hamster models. The results suggest that the compound could be exploited as an antileishmanial drug. PMID- 11937328 TI - Higher acyclic nitrogen containing deoxy sugar derivatives: a new lead in the generation of antimycobacterial chemotherapeutics. AB - Syntheses of higher acyclic nitrogen containing deoxy sugar derivatives via nitroaldol reaction of different nitroalkanes with 2,3-dideoxy-alpha,beta unsaturated aldehydo sugars obtained from glycals namely acetylated glucal and galactal and their in vitro antimycobacterial activity are presented. PMID- 11937329 TI - Determination of a sugar chain and its linkage site on a glycoprotein TIME-EA4 from silkworm diapause eggs by means of LC-ESI-Q-TOF-MS and MS/MS. AB - The electrospray ionization (ESI)-tandem quadrupole/orthogonal-acceleration time of-flight (Q-TOF) mass spectrometer combined with the nano-HPLC system was utilized to determine the glycosylation site and the glycan structure in glycoprotein TIME-EA4 (EA4) from Bombyx diapause eggs. LC-MS analysis of EA4 and deglycosylated EA4 indicated that the carbohydrate moiety of EA4 has the mass of 730.58 Da. Then, EA4 was digested with trypsin and chymotrypsin to identify the glycosylated peptide. The peptide fragment from G1y21 to Phe25 was found to carry the carbohydrate moiety. LC-MS/MS analysis of this peptide fragment revealed the sequence of the attached oligosaccharide and the glycosylation site at the same time. The present methodology utilizing the combination of the nano-HPLC system and a highly sensitive Q-TOF mass spectrometer is demonstrated to be quite effective for analyses of glycoproteins of relatively low purity and limited availability from natural sources. PMID- 11937330 TI - Metabolites of orally active NO-independent pyrazolopyridine stimulators of soluble guanylate cyclase. AB - The pyrazolopyridine stimulators of soluble guanylate cyclase BAY 41-2272 and 41 8543 were oxidised in rats and dogs at their 5-pyrimidinyl-cyclopropyl and morpholino residue. These metabolites activate the soluble guanylate cyclase, induce vasoelaxation and thereby may contribute to the in vivo activity of BAY 41 2272 and BAY 41-8543. PMID- 11937331 TI - Comparison of the inhibition of human and Trypanosoma cruzi prolyl endopeptidases. AB - Prolyl endopeptidases (PEPs) have been found in numerous species. Inhibitors of human enzyme could correct cognitive deficits in Alzheimer patients while inhibition of Trypanosoma cruzi PEP could prevent invasion phase in Chagas disease. A structure-activity relationship study carried out in a tetrahydroisoquinoline series allowed to obtain potent competitive inhibitors superior to SUAM-1221. Besides, inhibitors expected to act according to an irreversible mechanism revealed to be superior to JPT-4819, for applications linked to human enzyme inhibition. PMID- 11937332 TI - Arbutin synthase, a novel member of the NRD1beta glycosyltransferase family, is a unique multifunctional enzyme converting various natural products and xenobiotics. AB - Plant glucosyltransferases (GTs) play a crucial role in natural product biosynthesis and metabolization of xenobiotics. We expressed the arbutin synthase (AS) cDNA from Rauvolfia serpentina cell suspension cultures in Escherichia coli with a 6 x His tag and purified the active enzyme to homogeneity. The recombinant enzyme had a temperature optimum of 50 degrees C and showed two different pH optima (4.5 and 6.8 or 7.5, depending on the buffer). Out of 74 natural and synthetic phenols and two cinnamyl alcohols tested as substrates for the AS, 45 were accepted, covering a broad range of structural features. Converting rates comparable to hydroquinone were not achieved. In contrast to this broad acceptor substrate specificity, only pyrimidine nucleotide activated glucose was tolerated as a donor substrate. Nucleotide and amino acid sequence analysis revealed AS to be a new member of the NRD1beta family of glycosyl transferases and placed the enzyme into the group of plant secondary product GTs. Arbutin synthase is therefore the first example of a broad spectrum multifunctional glucosyltransferase. PMID- 11937333 TI - Design and synthesis of a selective EP4-receptor agonist. Part 3: 16-phenyl-5 thiaPGE(1) and 9-beta-halo derivatives with improved stability. AB - To identify a new selective EP4-agonist with improved chemical stability, further chemical modification of those reported previously was continued. We focused our attention on chemical modification of the alpha chain of 3,7-dithiaPGE(1) and selected 5-thiaPGE(1) as a new chemical lead. Introduction of an optimized omega chain to the 5-thiaPG skeleton afforded m-methoxymethyl derivative 33a, which showed the most potent EP4-receptor agonist activity and good subtype-selectivity both in vitro and in vivo. 9beta-HaloPGF derivatives were also synthesized and biologically evaluated in an attempt to block self-degradation of the beta hydroxyketone moiety. Among these series, and 39b showed potent agonist activity and good subtype-selectivity. Structure-activity relationships (SARs) are also discussed. PMID- 11937334 TI - Study on quantitative structure-toxicity relationships of benzene derivatives acting by narcosis. AB - Hydrophobicity (logP) as well as quantiative structure-toxicity relationships (QSTRs) of some benzene derivatives acting by narcosis have been established based on narcotic mechanisms of action and toxicity data to the fathead minnow, Daphnia magna and Vibrio fischeri using information-theoretic topological index (Id). Excellent results are obtained in multiparametric regression upon introduction of dummy parameters (indicator variables). Consistent increase in R(2)(A) values indicated that inspite of collinarity between Id and one of the indicator variables (I(3-6)) the proposed models are statistically significant. PMID- 11937335 TI - Molecular recognition by Kluyveromyces of amphotericin B-loaded, galactose tagged, poly (lactic acid) microspheres. AB - In an effort to develop a new way of drug delivery, especially for polyenic antifungal molecules, we have incorporated amphotericin B (AmB) into biodegradable galactosylated poly (L-lactic acid) (L-PLA) and poly (L-lactic-co glycolic acid) (PLGA) microspheres. These drug carriers were prepared by solvent evaporation using an oil/water (o/w) emulsion. The ratio of galactosyl spacers with different chain lengths was 1.74-2.78%. The maximal quantity of AmB encapsulated reported to 100 mg of the galactosylated microspheres was 7.14 mg for L-PLA (encapsulation rate 45% of mole) and 6.42 mg for PLGA derivatives (encapsulation rate 81% of mole). In our yeast model, drug release depended on three factors: (i) presence of galactosylated antennae, (ii) length of galactosyl antenna and (iii) nature of the polymer. More of the AmB trapped in PLGA microspheres was released than from PLA microspheres. These novel functionalised microspheres could be required for the delivering of therapeutic agents according to their recognition to specific cells. PMID- 11937336 TI - Synthesis of (1S,2R)-1-phenyl-2-[(S)-1-aminopropyl]-N,N diethylcyclopropanecarboxamide (PPDC) derivatives modified at the carbamoyl moiety as a new class of NMDA receptor antagonists. AB - (1S,2R)-1-Phenyl-2-[(S)-1-aminopropyl]-N,N-diethylcyclopropanecarboxamide (PPDC, ), which is a conformationally restricted analogue of the antidepressant milnacipran [(+/-)-1], represents a new class of potent NMDA receptor antagonists. A series of PPDC analogues modified at the carbamoyl moiety were synthesized. Among these, (1S,2R)-1-phenyl-2-[(S)-1-aminopropyl]-N,N dipropylcyclopropanecarboxamide (4d) was identified as the most potent NMDA receptor antagonist in this series and clearly reduced the MMDA receptor mediated potentiation of rat hippocampal slices, a model of long-term potentiation (LTP). The three-dimensional structure of 4d was also analyzed in detail to clarify the receptor-binding conformation. PMID- 11937337 TI - Synthesis and biological evaluation of aroylguanidines related to amiloride as inhibitors of the human platelet Na(+)/H(+) exchanger. AB - Pyridine and benzene bioisosteres of amiloride were synthesized and evaluated for their inhibitory potency against the sodium-hydrogen exchanger (NHE) involved in intracellular pH regulation. The inhibition of NHE was determined by using the platelet swelling assay (PSA) in which the swelling of human platelets was induced by their incubation in an acid buffer (pH 6.7). Additionally, the inhibitory potency of the most active compounds was assessed by measuring the inhibition of the EIPA-sensitive (22)Na(+) uptake (UIA) by human platelets after intracellular acidosis. The results indicated that several benzene derivatives and compounds bearing an carbonylguanidine moiety in the meta position of the pyridine nitrogen were much more potent than amiloride (PSA:IC(50)=43.5 microM; UIA:IC(50)=100.1 microM), but less than EIPA, a pyrazine NHE inhibitor (PSA:IC(50)=0.08 microM; UIA:IC(50)=0.5 microM). In both biological assays (2 amino-5-bromo-pyridine-3-carbonyl)guanidine (32) was the most active molecule (PSA: IC(50)=0.8 microM, UIA : IC(50)=0.8 microM). Our investigations demonstrated that the replacement of the pyrazine ring of amiloride by a pyridine or a phenyl ring improved the NHE inhibitory potency (phenyl >pyridine >pyrazine). PMID- 11937338 TI - Design and synthesis of novel tetra-peptide motilin agonists. AB - A series of novel tetra-peptide motilin agonists, having the general structure H Phe-Val-X-Ile-NH(2), were designed, on the basis of structure-activity relationship studies of motilin. Peptides, in which X is a side chain substituted tryptophan residue, have agonistic activity. H-Phe-Val-Trp(2'-CH(2)CH(2)OH)-Ile NH(2)(7), H-Phe-Val-Trp(2'-SCH(3))-Ile-NH(2)(8), and H-Phe-Val-Trp(2' SCH(2)CH(2)CH(3))-Ile-NH(2)(9), showed an EC(50) for contractile activity in the rabbit smooth muscle of 14.1+/-3.2, 12.9+/-4.1, and 4.6+/-1.6 microM, respectively. Interaction of the tryptophan aliphatic side chain with motilin receptor appears to influence the signal transduction via motilin receptor. PMID- 11937339 TI - Efficient and highly stereoselective synthesis of a beta-lactam inhibitor of the serine protease prostate-specific antigen. AB - An efficient synthesis of a beta-lactam precursor of the serine protease, prostate-specific antigen inhibitor 1 has been accomplished. The synthesis relies on two completely stereoselective reactions that allow the introduction of the stereocenters at C-3 and C-4 of the azetidinone ring in a predictable manner. PMID- 11937340 TI - 2-O-substituted cyclodextrins as reversal agents for the neuromuscular blocker rocuronium bromide. AB - A series of secondary face modified cyclodextrins (CDs) were synthesised with the aim of constructing host molecules capable of forming host-guest complexes with neuromuscular blockers, especially with rocuronium bromide. Perfacial 2-O substitution of gamma-CD with 4-carboxybenzyl resulted in a CD host molecule 1 that forms a 1:1 binary complex with rocuronium bromide (K(a) 6.2 x 10(5) M(-1)). The biological activities of this compound and other derivatives as reversal agents of rocuronium bromide were examined in vitro (mouse hemi-diaphragm) and in vivo (anaesthetized guinea pigs). The host molecule 1 was found to exert potent reversal activity (ED(50) 0.21 micromol/kg, iv) against rocuronium-induced neuromuscular block, and thus proved the viability of using host molecules as antidotes of a biologically active compound. PMID- 11937341 TI - Novel morpholinone-based D-Phe-Pro-Arg mimics as potential thrombin inhibitors: design, synthesis, and X-ray crystal structure of an enzyme inhibitor complex. AB - A morpholinone structural motif derived from D(+)- and L(-)-malic acid has been used as a mimic of D-Phe-Pro in the thrombin inhibiting tripeptide D-Phe-Pro-Arg. In place of Arg the more rigid P1 truncated p-amidinobenzylamine (Pab) or 2-amino 5-aminomethyl-3-methyl-pyridine have been utilized. The synthetic strategy developed readily delivers these novel thrombin inhibitors used to probe the alpha-thrombin inhibitor binding site. The best candidate in this series of thrombin inhibitors exhibits an in vitro IC(50) of 720 nM. The X-ray crystal structure of this candidate co-crystallized with alpha-thrombin is discussed. PMID- 11937342 TI - Syntheses, biological evaluation and QSAR study on antitumor activity of 1,5-N,N' disubstituted-2-(substituted benzenesulphonyl) glutamamides. AB - We have reported [unpublished data] the synthesis and QSAR of 5-substituted-2 (substituted benzenesulphonyl) glutamines which have shown the importance of steric factor on the aliphatic chain. N-Phthalyl isoglutamine, having the substitution at position 1 of the glutamic acid moiety, is the metabolite of recently approved thalidomide for different types of tumors by US FDA. Based on these, 36 new 1,5-N,N'-disubstituted-2-(substituted benzenesulphonyl) glutamamides were synthesized, as tools for further elucidation of the structural requirements for antitumor activity. All the synthesized compounds were tested for antitumor activity against Ehrlich Ascites Carcinoma (EAC) in Swiss albino mice using tumor weight as inhibitory parameter. Quantitative structure-activity relationship (QSAR) studies of these analogues revealed that the electron donating groups on the phenyl ring are found to be mandatory for the activity which was also proved by the negative coefficient of indicator parameter I(3,) for NO(2) group on the phenyl ring. Molecular volume (MV) and steric factor at R(5) position also plays a role in ligand-receptor interactions. PMID- 11937343 TI - Synthesis of feruloyl-myo-insitol derivatives and their inhibitory effects on phorbol ester-induced superoxide generation and Esptein-Barr virus activation. AB - We prepared 14 feruloyl-myo-inositol derivatives, and evaluated the relationships between their stereostructure and inhibitory activity toward the 12-O tetradecanoylphorbol-13-acetate (TPA)-induced superoxide (O(2)(-)) generation. And further, their suppressive effect on the TPA-induced Epstein-Barr virus (EBV) activation was examined in order to estimate their anti-carcinogenic potentials. Among the derivatives tested, 1,6-O-bis[3-(4'-hydroxy-3'-methoxyphenyl)-2 propenoyl]-myo-inositol (6b) showed an excellent suppressive activity on the O(2)(-) generation at a concentration of 20 microM. For the suppressive effects on the EBV activation, 2,4,6-O-tris[3-(4'-hydroxy-3'-methoxyphenyl)-2-propenoyl] myo-inositol 1,3,5-orthoformate (9b) showed the highest activity at a concentration of 100 microM among the derivatives tested. These results suggest that the inhibitory potencies of feruloyl-myo-inositol derivatives depend on the stereostructure of molecules rather than the hydrophobicity of molecules. PMID- 11937344 TI - Synthesis and structure-activity relationship of diarylamide urea derivatives as selective inhibitors of the proliferation of human coronary artery smooth muscle cells. AB - A series of diarylamide urea derivatives were synthesized and evaluated for their inhibitory activities against human coronary artery smooth muscle cells (SMCs) and human coronary artery endothelial cells (ECs). Compound 2o was superior to the lead compound, Tranilast, in terms of its potency of the inhibitory activity and cell selectivity. PMID- 11937345 TI - Structure-activity relationships of seco-prezizaane terpenoids in gamma aminobutyric acid receptors of houseflies and rats. AB - Thirteen seco-prezizaane terpenoids isolated from star anise species (Illcium floridanum, Illcium parviflorum, and Illcium verum) were investigated for their ability to inhibit the specific binding of [(3)H]4'-ethynyl-4-n propylbicycloorthobenzoate (EBOB), a non-competitive antagonist of gamma aminobutyric acid (GABA) receptors, to housefly-head and rat-brain membranes. Veranisatin A was found to be the most potent inhibitor in both membranes, with an IC(50)(fly) of 78.5 nM and an IC(50)(rat) of 271 nM, followed by anisatin (IC(50)(fly)=123 nM; IC(50)(rat)=282 nM). Six of the other 11 tested compounds were effective only in housefly-head membranes. Pseudoanisatin proved to display a high (>26-fold) selectivity for housefly versus rat GABA receptors (IC(50)(fly)=376 nM; IC(50)(rat) >10,000 nM). Although pseudoanisatin does not structurally resemble EBOB, Scatchard plots indicated that the two compounds bind to the same site in housefly receptors. Anisatin and pseudoanisatin exhibited moderate insecticidal activity against German cockroaches. Comparative molecular field analysis (CoMFA), a method of three-dimensional quantitative structure activity relationship (3D-QSAR) analysis, demonstrated that seco-prezizaane terpenoids can bind to the same site as do picrotoxane terpenoids such as picrotoxinin and picrodendrins, and the CoMFA maps allowed us to identify the parts of the molecules essential to high activity in housefly GABA receptors. PMID- 11937346 TI - A practical synthesis and biological evaluation of 9-halogenated PGF analogues. AB - A series of 9-halo PGF analogues 1-2 and 5-13 were synthesized and biologically evaluated. Among the compounds, 2 was the best EP2-receptor agonist. A practical method of synthesizing 2 via the Julia olefination of an aldehyde 3 with an optically active sulfone 4, which was prepared by Sharpless asymmetric epoxidation of 15, was developed. Other 9-halogenated PGF analogues were synthesized essentially by the same procedure and evaluated. The absolute configuration of 16-OH of 2 was determined as S by the X-ray analysis of a salt consisting of a 1/1 molar ratio of 2 and L-lysine. PMID- 11937347 TI - Synthesis and antimicrotubule activity of combretatropone derivatives. AB - Combretatropone is a hybrid of combretastatin and colchicine in which the o methoxyphenol of dihydrocombretastatin A-4 is replaced by an alpha methoxytropone. Derivatives of combretatropone have been synthesized and evaluated for antimicrotubule activity. All combretatropones were less active than the corresponding colchicine derivatives, supporting the idea that loss of ligand conformational entropy upon tubulin binding results in decreased potency for colchicinoid ligands. The structure-activity relationship of the combretatropone series was different than that of the colchicine series. These data indicate that conformationally mobile and conformationally rigid colchicinoids do not interact with the receptor site in the same manner. PMID- 11937348 TI - Discovery of 4,5-diphenyl-1,2,4-triazole derivatives as a novel class of selective antagonists for the human V(1A) receptor. AB - In the search for a novel class of selective antagonists for the human V(1A) receptor, high-throughput screening (HTS) of the Yamanouchi chemical library using CHO cells expressing the cloned human V(1A) (hV(1A)) receptor led to the discovery of 5-(4-biphenyl)-4-(2-methoxyphenyl)-3-methyl-1,2,4-triazole (3) which possessed the novel 4,5-diphenyl-1,2,4-triazole structure. Subsequent structure activity relationships studies on a series of the 4,5-diphenyl-1,2,4-triazole derivatives related to 3 revealed that the 4,5-diphenyl-1,2,4-triazole structure played an essential role in exerting high affinity for the hV(1A) receptor and that introduction of a basic amine moiety to the methoxy part of the 4-phenyl ring was effective in the improvement of both affinity for the hV(1A) receptor and selectivity versus the hV(2) receptor. Compound 3 and the 2 (morphorino)ethoxy derivative (11b) were shown to be antagonists for the hV(1A) receptor, from their effects on AVP-induced [Ca(2+)](i) response in CHO cells expressing the hV(1A) receptor. PMID- 11937349 TI - Vomilenine reductase--a novel enzyme catalyzing a crucial step in the biosynthesis of the therapeutically applied antiarrhythmic alkaloid ajmaline. AB - Delineation of the biochemical pathway leading to the antiarrhythmic Rauvolfia alkaloid ajmaline has been an important target in biosynthetic research for many years. The biosynthetic sequence starting with tryptamine and the monoterpene secologanin consists of about 10 different steps. Most of the participating enzymes have been detected and characterized previously, except those catalyzing the reduction of the intermediate vomilenine. A novel NADPH-dependent enzyme that reduces the intermediate has been isolated from Rauvolfia serpentina cell suspension cultures. Vomilenine reductase (M(r )43 kDa, temp opt 30 degrees C, pH opt 5.7-6.2), saturates the indolenine double bond of vomilenine with stereospecific formation of 2beta(R)-1,2-dihydrovomilenine. The described detection, enrichment and properties of the reductase not only closes a gap in ajmaline biosynthesis but is also a prerequisite for overexpressing the protein heterologously for final clarification of its molecular properties. PMID- 11937350 TI - Photo-control of nitric oxide synthase activity using a caged isoform specific inhibitor. AB - Nitric oxide (NO) plays a critical role in a number of physiological processes and is produced in mammalian cells by nitric oxide synthase (NOS) isozymes. Because of the diverse functions of NO, pharmaceutical interventions which seek to abrogate adverse effects of excess NOS activity must not interfere with the normal regulation of NO levels in the body. A method has been developed for the control of NOS enzyme activity using the localized photochemical release of a caged isoform-specific NOS inhibitor. The caged form of an iNOS inhibitor has been synthesized and tested for photosensitivity and potency. UV and multiphoton uncaging were verified using a hemoglobin-based assay. IC(50) values were determined for the inhibitor (70+/-11 nM), the caged inhibitor (1098+/-172 nM), the UV uncaged inhibitor (67+/-26 nM) and the multiphoton uncaged inhibitor (73+/ 11 nM). UV irradiation of the caged inhibitor resulted in a 86% reduction in iNOS activity after 5 min. Multiphoton uncaging had an apparent first order time constant of 0.007+/-0.001 min(-1). A therapeutic range exists, with molar excess of inhibitor to enzyme from 3- to 7-fold, over which the full dynamic range of the inhibition can be exploited. PMID- 11937351 TI - N-3(9)-arylpropenyl-N-9(3)-propionyl-3,9-diazabicyclo[3.3.1]nonanes as mu-opioid receptor agonists. Effects on mu-affinity of arylalkenyl chain modifications. AB - Two series of N-3-arylpropenyl-N-9-propionyl-3,9-diazabicyclo[3.3.1]nonanes (1b j) and of the reverted N-3-propionyl-N-9-arylpropenyl isomers (2b-j) as analogues of the previously reported analgesic N-3(9)-cinnamyl-N-9(3)-propionyl-3,9 diazabicyclo[3.3.1]nonanes (DBN) (1a, 2a) were synthesised and their affinity and selectivity towards opioid mu-, delta- and kappa-receptors were evaluated. Several compounds (1e,i,j-2d,e,f,g,j) exhibited a mu-affinity in the low nanomolar range with moderate or negligible affinity towards delta- and kappa receptors. The representative term N-9-(3,3-diphenylprop-2-enyl)-N-3-propionyl DBN (2d) displayed in vivo (mouse) a potent analgesic effect (ED(50) 3.88 mg/kg ip) which favourably compared with that of morphine (ED(50) 5 mg/kg ip). In addition, 2d produced in mice tolerance after a period twice as long with morphine. PMID- 11937352 TI - Major histocompatibility complex class II binding characteristics of peptoid peptide hybrids. AB - The major histocompatibility complex (MHC) class II binding requirements for solvent-exposed peptide residues were systematically studied using amino acid and peptoid substitutions. In a peptoid residue, the side chain is present on the backbone nitrogen atom as opposed to the alpha-carbon atom in an amino acid residue. To investigate the effect of this side chain shifting on MHC binding, three amino acids in the central part of the peptide sticking out of the binding groove were replaced by corresponding peptoid residues. Two peptoid-peptide hybrids showed large affinity decreases in the MHC-peptide binding assay. To investigate this affinity loss, the individual contributions to MHC binding affinity of the side chain (position), the putative hydrogen bond, and the flexibility were dissected. We conclude that the side chain position as well as the backbone nitrogen atom hydrogen bonding features of solvent-exposed residues in the peptide can be important for MHC binding affinity. PMID- 11937353 TI - Efficient synthesis of 'redox-switched' naphthoquinone thiol-crown ethers and their biological activity evaluation. AB - Series of naphthoquinone thiol-crown ethers had been prepared. The antibacterial, antifungal, and cytotoxic activities of these synthetic naphthoquinone thiol crown ethers were investigated. All of the compounds tested displayed antibacterial, cytotoxic and antifungal activities. The bis-naphthoquinone thiol crown ether 7a was the most potent inhibitor among tested analogues against Staphylococcus aureus methicillin resistance with MIC value of 2.68 microM. PMID- 11937354 TI - Potent DNA photocleavage by zinc(II) complexes of cationic bis-porphyrins linked with aliphatic diamine. AB - We have prepared zinc(II) complexes of cationic bis-porphyrins, as one of the attempts to improve less DNA photocleavage activities of the metal-free bis porphyrins composed of two H(2)TMPyP-like chromophores, linked with a series of aliphatic diamines. The less activities seemed to be derived from their intermolecular self-aggregation properties in aqueous solution. The zinc(II) insertion into the metal-free cationic bis-porphyrins completely removed their self-aggregation properties, most probably due to steric hindrance between axial ligands of zinc(II) chromophores of the cationic bis-porphyrins. The DNA photocleavage activities of the zinc(II) complexes were fully enhanced, which were three times larger than that of the lead compound H(2)TMPyP. Quantitative analysis of singlet oxygen production by photosensitization of cationic bis porphyrins was performed using 1,3-diphenylisobenzofuran, and the singlet oxygen productivities of them were found to be related to their solution properties. There is a good relationship between the activities and the productivities, which will provide insights into the further development of more effective DNA photocleavage agents. PMID- 11937355 TI - Linckosides A and B, two new neuritogenic steroid glycosides from the Okinawan starfish Linckia laevigata. AB - Two new steroid glycosides, named linckosides A and B, were isolated from the Okinawan starfish Linckia laevigata, and their stereostructures were elucidated by spectroscopic methods and chemical derivatization. Each of them possesses two monosaccharide units at C-3 and C-29 of a polyhydroxylated steroid aglycon. These steroid glycosides showed not only notable neuritogenic activity against PC12 cells but also significant synergistic effects on the NGF-induced neuronal differentiation of PC12 cells. Linckoside B showed higher activities than those of linckoside A, though the structural difference is only the kind of a sugar. PMID- 11937356 TI - Chemical modification of the beta-glucocerebrosidase inhibitor N-octyl-beta valienamine: synthesis and biological evaluation of 4-epimeric and 4-O-(beta-D galactopyranosyl) derivatives. AB - N-Octyl-beta-valienemine (1), a potent beta-glucocerebrosidase inhibitor, was chemically transformed into two biologically interesting compounds: the 4-epimer, beta-galacto-type N-octyl-valienamine, and the 4-O-(beta-D-galactopyranosyl) derivative, a carba-lactosylceramide analogue. The former, interestingly, could be demonstrated to act as a very effective inhibitor (IC(50)=0.3 microM) of human beta-galactosidase. The latter exhibited moderate inhibitory activity (IC(50)=20 microM) against beta-glucocerebrosidase (mouse liver). PMID- 11937357 TI - New apratoxins of marine cyanobacterial origin from Guam and Palau. AB - Two collections of the marine cyanobacterium Lyngbya sp. from Guam and Palau that both afforded the potent cytotoxin apratoxin A (1) each yielded different structural analogues with lower degrees of methylation. The new apratoxins, termed apratoxins B (2) and C (3), were evaluated for their in vitro cytotoxicity along with semisynthetic E-dehydroapratoxin A (4) to identify key structural elements responsible for the cytotoxicity and to initiate SAR studies on this novel family of depsipeptides. All analogues 2-4 displayed weaker cytotoxicity than 1, but to different extents. While compound 3 closely approached the cytotoxicity of 1, compounds 2 and 4 exhibited significantly reduced activity, possibly also related to a conformational change. The 16S rRNA genes of the different apratoxin producers have partially been sequenced and compared, and other genetic differences are currently being revealed. PMID- 11937358 TI - Analysis of stereoelectronic properties, mechanism of action and pharmacophore of synthetic indolo[2,1-b]quinazoline-6,12-dione derivatives in relation to antileishmanial activity using quantum chemical, cyclic voltammetry and 3-D-QSAR CATALYST procedures. AB - Several indolo[2,1-b]quinazoline-6,12-dione (tryptanthrin) derivatives exhibited remarkable activity at concentrations below 100 ng/mL when tested against in vitro Leishmania donovani amastigotes. The in vitro toxicity studies indicate that the compounds are fairly well tolerated in both macrophage and neuronal lines. An analysis based on qualitative and quantitative structure-activity relationship studies between in vitro antileishmanial activity and molecular electronic structure of 27 analogues of indolo[2,1-b]quinazoline-6,12-dione is presented here by using a combination of semi-empirical AM1 quantum chemical, cyclic voltammetry and a pharmacophore generation (CATALYST) methods. A modest to good correlation is observed between activity and a few calculated molecular properties such as molecular density, octanol-water partition coefficient, molecular orbital energies, and redox potentials. Electron transfer seems to be a plausible path in the mechanism of action of the compounds. A pharmacophore generated by using the 3-D QSAR of CATALYST produced a fairly accurate predictive model of antileishmanial activity of the tryptanthrins. The validity of the pharmacophore model extends to structurally different class of compounds that could open new frontiers for study. The carbonyl group of the five- and six membered rings in the indolo[2,1-b]quinazoline-6,12-dione skeleton and the electron transfer ability to the carbonyl atom appear to be crucial for activity. PMID- 11937359 TI - Synthesis and in vitro characterization of a tissue-selective fullerene: vectoring C(60)(OH)(16)AMBP to mineralized bone. AB - A tissue-vectored bisphosphonate fullerene, C(60)(OH)(16)AMBP [4,4-bisphosphono-2 (polyhydroxyl-1,2-dihydro-1,2-methanofullerene[60]-61-carboxamido)butyric acid], designed to target bone tissue has been synthesized and evaluated in vitro. An amide bisphosphonate addend, in conjunction with multiple hydroxyl groups, confers a strong affinity for the calcium phosphate mineral hydroxyapatite of bone. Constant composition crystal growth studies indicate that C(60)(OH)(16)AMBP reduces hydroxyapatite mineralization by 50% at a concentration of 1 microM, following a non-Langmuirian mechanism. Parallel studies with C(60)(OH)(30) also indicate an affinity for hydroxyapatite, but at a reduced level (28% crystal growth rate reduction at 1 microM) compared with C(60)(OH)(16)AMBP. This study is the first to demonstrate that a fullerene-based material can be successfully targeted to a selected tissue as a step toward the development of such materials for medical purposes, in general. PMID- 11937360 TI - Oligomers of glycamino acid. AB - Glycamino acids, a family of sugar amino acids, are derivatives of C-glycosides that possesses a carboxyl group at the C-1 position and an amino group replacing one of the hydroxyl groups at either the C-2, 3, 4, or 6 position. We have prepared a series of glucose-type glycamino acids as monomeric building blocks: these are derivatives of 2-NH(2)-Glc-beta-CO(2)H 1, 3-NH(2)-Glc-beta-CO(2)H 2, 4 NH(2)-Glc-beta-CO(2)H 3, and 6-NH(2)-Glc-beta-CO(2)H 4 and constructed four types of homo-oligomers, beta(1-->2)-linked I, beta(1-->3)-linked II, beta(1-->4) linked III, and beta(1-->6)-linked IV, employing the well-established N-Boc and BOP strategy. CD and NMR spectral studies of these oligomers suggested that only the beta(1-->2)-linked homo-oligomer possessed a helical structure that seems to be predetermined by the linkage position. Homo-oligomers with beta(1-->2) linkages I and beta(1-->6)-linkages IV were also subjected to O-sulfation, and these O-sulfated oligomers were found to be able, in a linkage-specific manner, to effectively inhibit L-selectin-mediated cell adhesion, HIV infection, and heparanase activity without the anticoagulant activity associated with naturally occurring sulfated polysaccharides such as heparin. PMID- 11937361 TI - Insight into the stereochemistry in the inhibition of carboxypeptidase A with N (hydroxyaminocarbonyl)phenylalanine: binding modes of an enantiomeric pair of the inhibitor to carboxypeptidase A. AB - Both D- and L-isomers of N-(hydroxyaminocarbonyl)phenylalanine () were shown to have strong binding affinity towards carboxypeptidase A (CPA) with D- being more potent than its enantiomer by 3-fold (Chung, S. J.; Kim, D. H. Bioorg. Med. Chem. 2001, 9, 185.). In order to understand the reversed stereochemical preference shown in the CPA inhibition, we have solved the crystal structures of CPA complexed with each enantiometer of up to 1.75 A resolution. Inhibitor L- whose stereochemistry belongs to the stereochemical series of substrate binds CPA like substrate does with its carbonyl oxygen coordinating to the active site zinc ion. Its hydroxyl is engaged in hydrogen bonding with the carboxylate of Glu-270. On the other hand, in binding of D- to CPA, its terminal hydroxyl group is involved in interactions with the active site zinc ion and the carboxylate of Glu-270. In both CPA small middle dot complexes, the phenyl ring in is fitted in the substrate recognition pocket at the S(1)' subsite, and the carboxylate of the inhibitors forms bifurcated hydrogen bonds with the guanidinium moiety of Arg-145 and a hydrogen bond with the guanidinium of Arg-127. In the complex of CPA small middle dotD-, the carboxylate of the inhibitor is engaged in hydrogen bonding with the phenolic hydroxyl of the down-positioned Tyr-248. While the L- binding induces a concerted movement of the backbone amino acid residues at the active site, only the downward movement of Tyr-248 was noted when D- binds to CPA. PMID- 11937362 TI - Exploration of the DTrp-NMeLys motif in the search for potent somatostatin antagonists. AB - Previous studies from this laboratory demonstrated that N-methylation at Lys(5) residue in somatostatin octapeptide antagonist analogues increased the GH release inhibition potency by as much as 300%. We have now further investigated N methylation of this Lys(5) residue in conjunction with a number of N- and C terminal modifications previously found to give highly potent somatostatin receptor antagonists. Synthetic analogues were tested in a functional assay for their ability to inhibit somatostatin-inhibited GH release from rat pituitary cells in culture and to displace 125I-labeled somatostatin from CHO cells transfected with the five known human somatostatin receptors. Several interesting observations resulted from the study. Replacement of liphophilic Nal(8) at the C terminus with a hydrophilic His(8) resulted in the increased affinity and selectivity for type 2 receptor to give the most potent antagonist analogue yet discovered (K(i), 1.5 nM), although in the rat pituitary cells inhibitory activity on somatostatin inhibited GH release decreased somewhat. A His(3) substitution within the cyclic portion of the analogues retained pituitary cell potency and affinity for type 2 receptor as did substitution with Bip(8) and Fpa(1). Replacement of Cpa(1) with Iph(1) did not effect the affinity for type 2 receptor significantly, but did decrease the effects on rat cell GH release. Iph(3) within-ring substitution increased the selectivity for sst(2) appreciably although the affinity for that receptor was considerably decreased. Substitution of Npa(3) resulted in good selectivity for sst(2) receptor. Replacement of Nal(8) with D-Trp(8) also increased the selectivity for type 2 receptor. Use of a 'bivalent ligand' approach in which two peptides were joined by 4,4' biphenyldicarbonyl as a spacer destroyed the affinity for all the subtypes, however, the bivalent ligand formed with the Ahp spacer displayed significant affinity and high selectivity for the type 2 receptor. PMID- 11937363 TI - 3-Oxa-15-cyclohexyl prostaglandin DP receptor agonists as topical antiglaucoma agents. AB - A series of prostaglandin DP agonists containing a 3-oxa-15-cyclohexyl motif was synthesized and evaluated in several in vitro and in vivo biological assays. The reference compound ZK 118.182 (9beta-chloro-15-cyclohexyl-3-oxa-omega-pentanor PGF(2alpha)) is a potent full agonist at the prostaglandin DP receptor. Saturation of the 13,14 olefin affords AL-6556, which is less potent but is still a full agonist. Replacement of the 9-chlorine with a hydrogen atom or inversion of the carbon 15 stereochemistry also reduces affinity. In in vivo studies ZK 118.182 lowers intraocular pressure (IOP) upon topical application in the ocular hypertensive monkey. Ester, 1-alcohol, and selected amide prodrugs of the carboxylic acid enhance in vivo potency, presumably by increasing bioavailability. The clinical candidate AL-6598, the isopropyl ester prodrug of AL-6556, produces a maximum 53% drop in monkey IOP with a 1 microg dose (0.003% w/w) using a twice-daily dosing regime. Synthetically, AL-6598 was accessed from known intermediate 1 using a novel key sequence to install the cis allyl ether in the alpha chain, involving a selective Swern oxidative desilylation of a primary silyl ether in the presence of a secondary silyl ether. In this manner, 136 g of AL-6598 was synthesized under GMP conditions for evaluation in phase I clinical trials. PMID- 11937364 TI - Synthesis and SAR of N-benzoyl-L-biphenylalanine derivatives: discovery of TR 14035, a dual alpha(4)beta(7)/alpha(4)beta(1) integrin antagonist. AB - alpha(4)beta(1) and alpha(4)beta(7) integrins are key regulators of physiologic and pathologic responses in inflammation and autoimmune disease. The effectiveness of anti-integrin antibodies to attenuate a number of inflammatory/immune conditions provides a strong rationale to target integrins for drug development. Important advances have been made in identifying potent and selective candidates, peptides and peptidomimetics, for further development. Herein, we report the discovery of a series of novel N-benzoyl-L-biphenylalanine derivatives that are potent inhibitors of alpha4 integrins. The potency of the initial lead compound (1: IC(50) alpha(4)beta(7)/alpha(4)beta(1)=5/33 microM) was optimized via sequential manipulation of substituents to generate low nM, orally bioavailable dual alpha(4)beta(1)/alpha(4)beta(7) antagonists. The SAR also led to the identification of several subnanomolar antagonists (134, 142, and 143). Compound 81 (TR-14035; IC(50) alpha(4)beta(7)/alpha(4)beta(1)=7/87 nM) has completed Phase I studies in Europe. The synthesis, SAR and biological evaluation of these compounds are described. PMID- 11937365 TI - Synthesis and in vitro antitumor activity of thiophene analogues of 5-chloro-5,8 dideazafolic acid and 2-methyl-2-desamino-5-chloro-5,8-dideazafolic acid. AB - N-[5-[N-(2-Amino-5-chloro-3,4-dihydro-4-oxoquinazolin-6-yl)methylamino]-2 thenoyl]-L-glutamic acid (6) and N-[5-[N-(5-chloro-3,4-dihydro-2-methyl-4 oxoquinazolin-6-yl)methylamino]-2-thenoyl]-L-glutamic acid (7), the first reported thiophene analogues of 5-chloro-5,8-dideazafolic acid, were synthesized and tested as inhibitors of tumor cell growth in culture. 4-Chloro-5-methylisatin (10) was converted stepwise to methyl 2-amino-5-methyl-6-chlorobenzoate (22) and 2-amino-5-chloro-3,4-dihydro-6-methyl-4-oxoquinazoline (19). Pivaloylation of the 2-amino group, followed by NBS bromination, condensation with di-tert-butyl N-(5 amino-2-thenoyl)-L-glutamate (28), and stepwise cleavage of the protecting groups with ammonia and TFA yielded. Treatment of 9 with acetic anhydride afforded 2,6 dimethyl-5-chlorobenz[1,3-d]oxazin-4-one (31), which on reaction with ammonia, NaOH was converted to 2,6-dimethyl-5-chloro-3,4-dihydroquinazolin-4-one (33). Bromination of, followed by condensation with and ester cleavage with TFA, yielded. The IC(50) of and against CCRF-CEM human leukemic lymphoblasts was 1.8+/ 0.1 and 2.1+/-0.8 microM, respectively. PMID- 11937366 TI - Novel tacrine derivatives that block neuronal calcium channels. AB - A new series of tacrine (9-amino-1,2,3,4-tetrahydroacridine) derivatives were synthesized and their effects on 45Ca(2+) entry into bovine adrenal chromaffin cells stimulated with dimethylphenylpiperazinium (DMPP) or K(+), studied. At 3 microM, compound 1 did not affect (45)Ca(2+) uptake evoked by DMPP. Compounds 14, 15 and 17 inhibited the effects of DMPP by 30%. Compounds 3, 9 and tacrine blocked the DMPP signal by about 50%. Compounds 5 and 12 were the most potent blockers of DMPP-stimulated 45Ca(2+) entry (90%); the rest of the compounds inhibited the effects of DMPP by 70-80%. Compounds 1, 3, 4, 8, 10, 11, 13, 16, 17 and tacrine inhibited 45Ca(2+) uptake induced by K(+) about 20%. Compounds 6, 14 and 15 inhibited the K(+) effects by 10% or less. Compounds 7, 9, 12 and 18 blocked the K(+) signal by 30% and, finally, compounds 2 and 5 inhibited the K(+) induced 45Ca(2+) entry by 50%. None of the new compounds was as effective as diltiazem (IC(50)=0.03 microM) in causing relaxation of the rat aorta precontracted with 35 mM K(+); the most potent was compound 7 (IC(50)=0.3 microM). Compounds 5, 6, 8, 9, 10 and 13 had IC(50)s around 10 microM and compounds 3, 4, 11 and 12 around 20 microM. Blockade of Ca(2+) entry through neuronal voltage-dependent Ca(2+) channels, without concomitant blockade of vascular Ca(2+) channels, suggests that some of these compounds might exhibit neuroprotectant effects but not undesirable hemodynamic effects. PMID- 11937405 TI - Enough time wasted in South Africa. PMID- 11937407 TI - Home-grown Vietnamese cholera vaccine "completely safe". PMID- 11937408 TI - New antimalarial promises cheap and effective treatment. PMID- 11937410 TI - Satellites spy on infectious disease. PMID- 11937412 TI - Staphylococcal vaccine one step closer to reality. PMID- 11937413 TI - Brain tumour-JC virus link strengthened but not proven. PMID- 11937414 TI - Vaccine vanquishes varicella. PMID- 11937415 TI - Australian find suggests worldwide reach for metapneumovirus. PMID- 11937417 TI - Heading for disaster: HIV prevalence soars in eastern Europe. PMID- 11937418 TI - "Getting ahead of the curve": a strategy and its implementation. PMID- 11937419 TI - Quinine in the modern treatment of falciparum malaria. PMID- 11937420 TI - Dengue in Cuba: mobilisation against Aedes aegypti. PMID- 11937421 TI - Epidemiology of drug-resistant malaria. AB - Since the first reports of chloroquine-resistant falciparum malaria in southeast Asia and South America almost half a century ago, drug-resistant malaria has posed a major problem in malaria control. By the late 1980s, resistance to sulfadoxine-pyrimethamine and to mefloquine was also prevalent on the Thai Cambodian and Thai-Myanmar (Thai-Burmese) borders, rendering them established multidrug-resistant (MDR) areas. Chloroquine resistance spread across Africa during the 1980s, and severe resistance is especially found in east Africa. As a result, more than ten African countries have switched their first-line drug to sulfadoxine-pyrimethamine. Of great concern is the fact that the efficacy of this drug in Africa is progressively deteriorating, especially in foci in east Africa, which are classified as emerging MDR areas. Urgent efforts are needed to lengthen the lifespan of sulfadoxine-pyrimethamine and to identify effective, affordable, alternative antimalarial regimens. Molecular markers for antimalarial resistance have been identified, including pfcrt polymorphisms associated with chloroquine resistance and dhfr and dhps polymorphisms associated with sulfadoxine pyrimethamine resistance. Polymorphisms in pfmdr1 may also be associated with resistance to chloroquine, mefloquine, quinine, and artemisinin. Use of such genetic information for the early detection of resistance foci and future monitoring of drug-resistant malaria is a potentially useful epidemiological tool, in conjunction with the conventional in-vivo and in-vitro drug-sensitivity assessments. This review describes the various features of drug resistance in Plasmodium falciparum, including its determinants, current status in diverse geographical areas, molecular markers, and their implications. PMID- 11937422 TI - p values for pathogens: statistical inference from infectious-disease data. AB - Certain features of infectious-disease data-including the aggregated nature of the data, confounding variables, correlated variables, and non-linear relations complicate the use of standard statistical procedures. Using data on a helminth infection, we review the use of three parametric tests (analysis of variance, linear regression, and logistic regression), address the complications arising from violation of the assumptions for these tests, and suggest methods of correction. We also compare the relative merits of parametric methods with equivalent non-parametric approaches, and illustrate the differences produced with results from a Kruskal-Wallis test and a t test. The value of using a resampling method-bootstrapping-is also shown. Finally, we discuss problems arising from use of a study design that requires data on the same attribute to be collected from the same individual over a period of time, and present three methods for overcoming this complication, showing that, in the example used, the mixed effect model and generalised estimating equation give similar results. PMID- 11937423 TI - Monotherapy or aminoglycoside-containing combinations for empirical antibiotic treatment of febrile neutropenic patients: a meta-analysis. AB - We set out to compare the efficacy of antibiotic monotherapy with that of combination therapy including an aminoglycoside for empirical treatment of febrile neutropenic cancer patients. We did a meta-analysis of 29 randomised clinical trials pooling data from 4795 febrile episodes and a subset of 1029 bacteraemic episodes by both fixed and random effects models. Outcome measure was clinical failure of antibiotic treatment, defined as modification of the initially allocated regimen or death during treatment. In febrile episodes, the pooled odds ratio (OR) of clinical failure with monotherapy versus combination therapy was 0.88, with 95% CI from 0.78 to 0.99 by the fixed effects model, and 0.87 with 95% CI from 0.75 to 1.01 by the more conservative random effects model. For bacteraemic episodes, the pooled OR of failure with monotherapy was 0.70 (0.54 to 0.92) by the fixed effects model, and 0.72 (0.54 to 0.95) by the random effects model. We conclude that monotherapy has been as effective as aminoglycoside-containing combinations for empirical treatment of febrile neutropenia. PMID- 11937424 TI - Multiplexed immunoassays by flow cytometry for diagnosis and surveillance of infectious diseases in resource-poor settings. AB - An accurate, rapid and cost-effective diagnosis is the cornerstone of efficient clinical and epidemiological management of infections. Here we discuss the relevance of an emerging technology, multiplexed immunoassays read by flow cytometry, for the diagnosis of infectious diseases. In these assays, multiple fluorescent microspheres, conjugated to different antigens or antibodies, constitute the solid phase for detecting antibodies or antigens in biological samples. These assays seem to be more sensitive than traditional immunoassays, have a high throughput capacity, and provide a wide analytical dynamic range. Additionally, they have multiplexing ability-ie, they are capable of measuring multiple antibodies or antigens simultaneously. We discuss four different areas where this technology could make an impact in resource-poor settings: (i) infections causing rash and fever in children; (ii) sero-epidemiological studies on vaccine-preventable diseases; (iii) management of genital ulcers and vaginal discharge; and (iv) screening of infections in blood banking. We predict a widespread use for a new breed of small, affordable, practical flow cytometers as field instruments for replacing ELISA and RIA tests, which will also be capable of doing cellular immunological tests such as CD4+ T-cell enumeration and Plasmodium falciparum detection in whole blood. PMID- 11937425 TI - Sequencing the Aspergillus fumigatus genome. AB - Aspergillus fumigatus is the most common mould pathogen of human beings and unusually causes both invasive disease in immunocompromised patients and allergic disease in patients with atopic immune systems. 4% of patients dying in modern European teaching hospitals have invasive aspergillosis and it is the leading infectious cause of death in leukaemia and bone marrow transplant patients. Until 2001, only two licensed antifungal drugs were available to treat aspergillosis amphotericin B and itraconazole. Its 28-30Mb genome is being sequenced in an international collaboration, with the Wellcome Trust Sanger Institute (UK) and The Institute for Genomic Research (TIGR, USA) as the two main centres. A whole genome shotgun approach was adopted and initiated in 2001 with an expected completion date in 2003. The complete sequence will permit identification of pathways specific to pathogenic Aspergillus species, help identify new targets for antifungal drugs, and enable investigations into the basic biology of fungi. Numerous secondary metabolic pathways with biotechnological applications and pharmacological properties are found in the Aspergilli and the genome sequence will facilitate research in this area. PMID- 11937426 TI - Gary Maartens--committed to improving public health in South Africa. PMID- 11937429 TI - Secrets of the Bulgarian bacillus. PMID- 11937431 TI - Mind-body effect: insulinlike growth factor-1; clinical depression; and breast, prostate, and other cancer risk-an unmeasured and masked mediator of potential significance? AB - A possible relation may exist between higher insulinlike growth factor (IGF)-1 levels and the risk of premenopausal breast, prostate, or other cancers from recent prospective and case-control studies. Separately, a large prospective study has shown a potential association between chronic depression and cancer risk, whereas other preliminary studies have suggested a link between increasing IGF levels with major depression. Other studies have found that certain standard cancer treatments reduce the effect and levels of IGF, and 1 small study has found that standard antidepressants may reduce IGF levels in some patients diagnosed with major depression. It is possible that IGF-1 (or other IGFs) may be a mediator between depression and cancer, because separately both have been implicated in the risk of various cancers. This hypothesis first and foremost relies on the unsettled notion that an actual relation already exists or may be found in the future. It requires more extensive investigation because several studies have not found an association between IGF or depression and cancer risk, and other limitations such as small sample sizes and other influential factors on IGF levels need to be elucidated. Whether or not a mind-body effect exists with cancer, as seems to be the case with other diseases, remains to be seen. Is depression a potential confounder of human studies that attempts to establish a relation between IGF and disease? All of the studies to date have attempted to at least determine the effect of a variety of factors on IGF levels-including age, sex, diet, family history, weight, smoking status-and then have adjusted their results accordingly. Depression has not been included in this list of potential factors that may need to be considered when analyzing IGF-1 data and cancer risks. The time seems ripe to at least define further the relation, if any, between IGF-1 and depression. PMID- 11937432 TI - Selenium and vitamin E supplements for prostate cancer: evidence or embellishment? AB - Selenium and vitamin E are probably 2 of the most popular dietary supplements considered for use in the reduction of prostate cancer risk. This enthusiasm is reflected in the initiation of the Selenium and Vitamin E Chemoprevention Trial (SELECT). Is there sufficient evidence to support the use of these supplements in a large-scale prospective trial for patients who want to reduce the risk of prostate cancer? Results from numerous laboratory and observational studies support the use of these supplements, and data from recent prospective trials also add partial support. However, a closer analysis of the data reveals some interesting and unique associations. Selenium supplements provided a benefit only for those individuals who had lower levels of baseline plasma selenium. Other subjects, with normal or higher levels, did not benefit and may have an increased risk for prostate cancer. The concept that supplements reduce prostate cancer risk only in those at a higher risk and/or those with lower plasma levels of these compounds is supported by trials examining beta-carotene supplements. Smokers may be the only individuals who benefit, as has also been shown with vitamin E supplementation. In 4 recent prospective studies, vitamin E was found to reduce the risk of prostate cancer in past/recent and current smokers and those with low levels of this vitamin. Vitamin E supplements in higher doses (> or =100 IU) were also associated with a higher risk of aggressive or fatal prostate cancer in nonsmokers from a past prospective study. The dose of vitamin E in the SELECT trial (400 IU/day) is 8 times higher than what has been suggested to be effective (50 IU/day) by the largest randomized prospective trial in which the incidence rate of prostate cancer was used as an endpoint. Recent research also suggests that dietary vitamin E may be associated with a lower risk of prostate cancer than the vitamin E supplement. Additionally, recent results from all past cardiovascular prospective, randomized trials suggest that vitamin E shows little benefit for cardiovascular disease risk, especially at the dose being used in the SELECT trial. Other intriguing positive findings from past prospective studies of supplements suggest that aspirin and other nonsteroidal anti-inflammatory drugs have a role in reducing the risk of prostate cancer or other types of cancer (eg, colon cancer). It may be time to conduct a large costly trial to reconsider the use of selenium and vitamin E supplements for the reduction of prostate cancer risk. Some evidence for the use of these supplements exists, but serious embellishment of study findings may be leading to an inappropriate use of these supplements in a clinical setting. PMID- 11937433 TI - Complementary/alternative therapies for reducing hot flashes in prostate cancer patients: reevaluating the existing indirect data from studies of breast cancer and postmenopausal women. AB - Vasomotor hot flashes are a common problem in women who are postmenopausal or receiving antiestrogen treatment for breast cancer. Hot flashes are also a common problem after orchiectomy/luteinizing hormone-releasing hormone therapy, occurring generally in 50% to 66% of these men. Prescribed treatments for hot flashes for men on hormonal ablation treatment for prostate cancer are well documented. These conventional agents have shown good results, but their long term efficacy, safety, and cost are still questioned. Therefore, the search for other viable agents, including nontraditional treatments, continues. Complementary/alternative treatments to alleviate hot flashes in women have generated an enormous amount of interest. However, these options have received little attention in men with hot flashes. Research with vitamin E, soy, black cohosh, red clover, and numerous other alternative treatments in women may provide some indirect but valuable insight on their potential effectiveness in men. Many of these alternatives have been a disappointment in recent randomized trials of women, and it is likely that there will be similar results with men. However, numerous supplements have yet to be tested in a clinical trial against a placebo, and clinicians should become aware of this ever-increasing list. Patients should be made aware of the primary importance of lifestyle interventions that could partially affect hot flashes and immediately affect overall health, especially during the period of androgen suppression, when it is not uncommon to observe accelerated weight changes and insulin insensitivity. Otherwise, recent research with older and newer conventional agents, such as antidepressants or estrogen/progesterone, should be emphasized at this time for moderate-to-severe hot flashes that profoundly affect daily activities and/or sleep. Antidepressant supplements (St. John's wort) or acupuncture could also be an attractive option in future investigations. Low-dose estrogen seems particularly attractive, because it is inexpensive and may simultaneously reduce hot flashes and the risk of osteoporosis in men receiving long-term androgen suppression therapy; however, the potential for cardiovascular complications must be further investigated. Ultimately, adequate research (vs placebo) should determine the fate of the alternative supplements proposed for hot flash reduction. PMID- 11937435 TI - Is obesity a risk factor for prostate cancer, and does it even matter? A hypothesis and different perspective. AB - Measurement of obesity is not as simple as its definition. Currently, several methods of measuring obesity are used in clinical studies. Skinfold thickness, crude weight, lean body mass (LBM), body mass index (BMI), and waist-to-hip ratio (WHR) are some of the more popular methods, but each contains its inherent strengths and flaws. In general, the results of the largest studies on prostate cancer and obesity have not been conclusive. One of the largest studies found an inverse relation to prostate cancer in the youngest age groups. The age and duration of obesity or any rapid changes in weight gain, along with other unhealthy exposures, may have some relation to prostate cancer incidence and mortality. Early intrinsic or extrinsic exposure to estrogen or estrogenlike compounds may provide a protective effect. The timing and duration of a higher estrogen and/or lower testosterone exposure may have a beneficial or detrimental impact on the prognosis of an established prostate tumor. Negative exposures over time such as low levels of sex hormone-binding globulin (SHBG), a greater exposure to growth factors, elevated insulin levels, greater sympathetic activity, higher cholesterol levels, immune system dysfunction, inadequate diets, smoking status, and other factors may be associated with an increased risk of prostate cancer and other diseases. Obesity may also be associated with other cancers for similar and different reasons. For example, morbidity and mortality from postmenopausal breast cancer, colon, kidney, and other cancers are potentially associated with obesity. Other comorbidities such as cataracts, coronary heart disease, diabetes, erectile dysfunction, hypertension, and others are also associated with obesity. The 2 largest prospective studies on BMI and overall mortality have also demonstrated the substantial negative impact of excess weight on society. Prostate cancer risk and obesity need further research to establish if a true association exists, but at this time, does it really matter? Overall, the profound adverse effect of being obese on general health is dramatic, and this is what clinicians and patients need to remember. PMID- 11937434 TI - Complementary therapies for reducing the risk of osteoporosis in patients receiving luteinizing hormone-releasing hormone treatment/orchiectomy for prostate cancer: a review and assessment of the need for more research. AB - Osteoporosis in women has received a substantial amount of attention, but its impact in men is also significant and noteworthy. Those men who benefit from treatment for prostate cancer with androgen deprivation therapy (ADT) may also be at a higher risk for osteoporosis. Pharmacologic approaches to reduce this risk have received some attention. For example, agents such as bisphosphonates, estrogen receptor-binding drugs (diethylstilbestrol, tamoxifen, and raloxifene), calcitonin, and fluoride are some of the more promising interventions that have been previously outlined. In addition, statin drugs, or hepatic 3-hydroxy-3 methylglutaryl coenzyme A reductase inhibitors, have recently been hypothesized to lower osteoporosis risk. However, complementary therapies, which may also have an impact on reducing osteoporosis risk, have not received attention. Dietary and supplemental calcium and vitamin D have been shown, in some preliminary investigations, to maintain bone density in women and men. Numerous healthy and affordable dietary sources of this mineral and vitamin exist, and large intakes can be realistically achieved through proper education. Similarly, the supplemental dosages required to impact risk have been moderate, appear to be safe, are of low cost, and thus may provide an additional route for reducing risk, especially if these interventions are initiated at the start of medical treatment. More studies in men receiving ADT are needed because the existing work has mostly focused on men without castrate levels of male hormone. Additionally, many studies with conventional and nonconventional agents have only focused on individuals with baseline osteoporosis, rather than normal bone mineral densities or osteopenia. Other promising complementary therapies, such as weight-bearing exercise and abstaining from smoking, may also be of benefit. Newer estrogenic type supplements (eg, ipriflavone) appear interesting and have some preliminary data, but more research is desperately required to determine their actual impact and potential for adverse effects (such as lymphocytopenia from a recent trial). Simple, inexpensive, and potentially effective dietary and supplemental approaches to reduce the risk of osteoporosis in men exist, and they should be discussed with patients. Whether these approaches effectively reduce the risk of osteoporosis in men receiving androgen ablation remains to be determined. The possibility is intriguing, and future research is needed. In the meantime, it is important to keep in mind that these complementary approaches are, at the very least, an integral part of the conventional options used today to the reduce the risk of osteoporosis in men and women. PMID- 11937436 TI - Dietary fat reduction to reduce prostate cancer risk: controlled enthusiasm, learning a lesson from breast or other cancers, and the big picture. AB - Breast and prostate cancer share similar intrinsic and extrinsic risk factors. Based on laboratory, ecologic/international comparison, and case-control studies, the impact of dietary fat or other fat subtypes has been suggested as a potential route to reduce risk. Recent large-scale prospective studies have failed to find an association between fat and breast cancer risk. These studies may provide some insight for researchers examining the relation between fat and prostate cancer. Prospective studies to date have also failed to find a consistent association between prostate cancer and fat intake. Some fat subtypes (eg, saturated fat) or other lifestyle changes (eg, obesity, physical activity) may affect risk and progression of these cancers when examining the sum total of the research, but more precise and specific investigations in humans are needed to address these issues. Other concerns, such as the impact of excess energy or overall caloric consumption on carcinogenesis, still need to be addressed, as well as other methodologic limitations of past investigations. Large gaps exist in environmental (eg, diet, lifestyle) and heritable causes of these diseases. Regardless, until more extensive research is completed, lifestyle changes should be recommended based on reducing morbidity and mortality from cardiovascular disease-the number 1 cause of death in the United States. Additionally, cardiovascular disease remains the number 1 or 2 cause of death in patients diagnosed with breast or prostate cancer. Practical and simple dietary changes should be encouraged by health professionals because they could improve the overall longevity and quality of patients' lives. Numerous ongoing prospective studies of diet and cancer should provide researchers and the public with much needed answers in this area. PMID- 11937437 TI - [The importance of the summary]. PMID- 11937438 TI - [What is new in antithrombotic treatment?]. AB - Real progresses have been made during the past years in the comprehension of hemostasis mechanisms, along with rising of new antithrombotic drugs. The later include: 1) direct inhibitors of thrombin such as hirudin, bivalirudin, argatroban, melagatran and ximelagatran; 2) inhibitors of factor Xa such as the synthetic pentasaccharid and DX-9065a; 3) inhibitors of factor IXa; 4) inhibitors of tissue factor-factor VIIa complex such as tissue factor pathway inhibitor (TFPI) or NAPc2 (nematode anticoagulant peptide); 5) drugs enhancing endogenous anticoagulant activity, such as protein C or activated protein C; 6) drugs modulating endogenous fibrinolytic activity. These new drugs are promising a real decrease in mortality and morbidity due to venous thrombo-embolic disease, which is considered as a public health issue. Both physicians and biologists are concerned by these new antithrombotic agents, the former to think about new treatment strategies, the later to monitore, if necessary, the effects of such new drugs. Our review does not include antiplatelet agents which are indicated only in arterial thrombosis. PMID- 11937439 TI - [Tumor-induced immunosuppression]. AB - Tumor immunology is based on two essential concepts: immune surveillance, which implicate the host immune reactions against tumor cells, and tumor immune escape, which refers to the tumor-cell evasion process against the host immune system. The notion that a deficit in immune cell functions permits tumor growth has received experimental support with the discovery of several different biochemical defects in T lymphocytes that infiltrate cancers. Furthermore, expression of self antigens on the tumor surface impose potential barriers to the development of effective immune response. Tumors are able to overcome immune surveillance by changing the polarity of effectors cells, thus down-regulating the proliferation of tumor-specific cytotoxic T cells, or altering the effector compositions of immune cells within the tumor milieu, or both. Understanding the interaction between cancer cells and host immune cells is of importance for clinical applications or immunotherapy in cancer treatment. PMID- 11937440 TI - [Evaluation of portable point of care instrument: the i-Stat . Report of 7,000 analyses]. AB - To validate the use of the i-STAT portable point-of-care instrument, performance checks have been conducted to verify its accuracy for PCO2/PO2 (tonometry) and precision for pH/PCO2/PO2 (control solutions). Results obtained with the i-STAT (pH/PCO2/PO2/Hct) were also compared to those provided by the Bayer Rapidlab 865 routinely used in our laboratory. All these measurements have been conducted under the same conditions using three types of i-STAT cartridges (G3, EG6, EG7) with two of different life duration to each cartridge with a view to observing their potential variability between types and any effect that ageing may have on analytical performances. Differences observed in this respect are not clear enough to draw firm conclusions. On the other hand, we consider the level of PCO2 and particularly PO2 performances not sufficient to warranty their use in the absence of a very strict quality control coupled with a highly developed clinical sense. No real use can be made of values noted for hematocrit (1998 survey). PMID- 11937441 TI - [Screening of tetrahydrobiopterin deficiency among hyperphenylalaninemic patients]. AB - Tetrahydrobiopterin deficiency in hyperphenylalaninemic babies has to be rapidly recognized since the disease requires a specific follow-up. Based on specimen collection on filter paper, a simple strategy for the screening of this condition has been used since 1987. Urine pteridine measurement can detect 6-pyruvoyl tetrahydropterin synthase, GTPcyclohydrolase I and pterin-4a-carbinolamine dehydratase deficiencies and direct enzyme measurement in dried blood sample detects dihydropteridine-reductase deficiency. A total of 1,814 hyperphenylalaninemic patients have been studied and 34 tetrahydrobiopterin deficiencies have been detected. The strategy must commend itself by its convenience and simplicity, and can be use on all babies with hyperphenylalaninemia screened in the neonatal period, whatever their blood phenylalanine level. PMID- 11937442 TI - [Names-Lab : a model for the standardization of biology message exchanges]. AB - The growing requirements of communication between health partners urges to standardize the language used in laboratories data management systems. Names-Lab is the only semantic reference system allowing to describe synchronously the body of prescription and result messages. Its hierarchical structure allows a simple coding, easily accessible to all biologists without imposing or even requiring a language. Other internationally proposed systems as Loinc and Euclides are similar to Names-Lab in their purposes but are completely different by their content and structure. This prompts the medical and biologic community for a comparison of these tools before setting the choice of an international referencing system. PMID- 11937443 TI - [Interpretation of the CA125 kinetics during first line chemotherapy of the ovarian cancer: methodological aspects and characteristic profiles]. AB - Mathematical analysis of CA125 kinetics during first line chemotherapy allows calculation of various biologic parameters which are powerful indicators of the therapeutic efficiency. The purpose of this study is to present an original method of interpretation of CA125 kinetics based on both CA125 profile and its half-life value. The first part of this study reviews the practical modalities of CA125 kinetics analysis, the methods of calculation of the biologic parameters as well as the guidelines of interpretation. The second part of this work is dedicated to the presentation of CA125 profile characteristics in responders to chemotherapy, partially or totally nonresponders to chemotherapy, tumoral growth under treatment and tumor lysis syndrome. PMID- 11937444 TI - [Emerging parasitosis ans mycosis: risk and threats for the new millenium]. AB - For multiple reasons, the emergent infectious risks do not stop increasing these last twenty years. The climatic modifications and the human interventions modifying the biotope as well as the rapid spreading of resistant strains to treatments, generate re-emergence or emergence, all the more dramatic as the means of fight are reduced. These emergent or re-emergent diseases are extremely worrisome as their diagnosis and their prevention are often difficult. The important infesting power of parasites and the particularly effective capacities of adaptation of these eucaryotes contributed to the public health problems. Anthropozoonoses and zoonoses constitute a permanent risk the control of which is imaginary. The new pathogenic agents, the unusual clinical demonstrations in the context of deficiency of the host immune functions imply attentiveness and a permanent up to date of the knowledge of the biologist and of the different professionals of health. The risks with which are confronted the humanity during this century underline the necessity of determining mechanisms involved in the pathogenesis. The determination of the specific and vital biologic processes for the microorganism, could allow to define the most appropriated targets and the most effective and original means of fight. PMID- 11937445 TI - [Congenital human cytomegalovirus infection: value of human cytomegalovirus DNA quantification in amniotic fluid]. AB - A quantitative PCR assay (RS Elosa CMV, Lambdatech) was used to quantitate HCMV DNA in maternal amniotic fluid of 12 fetuses with congenital infection (group 1) and of 10 fetuses without congenital infection (group 2). HCMV detection was performed for both groups using culture and qualitative PCR. Histologic examinations of fetal tissues and placenta were carried out for 9 patients from group 1. The amniotic fluid viral loads were negative in all patients of group 2. In group 1, all viral loads were high (from 1.105 to > 107 cop/mL) and no difference was observed between symptomatic and asymptomatic foetuses. Further evaluation on larger samples is needed to define more precisely the pronostic value of HCMV DNA quantification in amniotic fluid. PMID- 11937446 TI - [Use of contrast phase microscopy in the evaluation of urine cytology]. PMID- 11937447 TI - [Malaria-related cytopenia]. PMID- 11937448 TI - [A case of oropharyngeal tularemia from duck plumage]. PMID- 11937449 TI - [Evaluation of HbA1c immunoassay on MIRA Plus analyseur]. PMID- 11937450 TI - [Evaluation of the Strep B OIA, a rapid identification test of group B streptococcal colonization of pregnant woman]. PMID- 11937451 TI - [Acute myologenous leukemia with skin involvement]. PMID- 11937452 TI - [News from the scientific committee of the SFBC]. PMID- 11937453 TI - [To diagnose or to exclude vitamine B12 deficiency]. PMID- 11937455 TI - Right heart support during off-pump coronary artery bypass surgery--a multi center study. AB - BACKGROUND: Off-pump coronary artery bypass (OPCAB) surgery for posterior and inferior wall vessels requires heart displacement, which leads to hemodynamic instability. Based on results indicating that displacement primarily alters right heart function, this study evaluates the safety and efficacy of right heart support during OPCAB surgery to the posterior and inferior wall vessels. METHODS: In a multi-center, prospective study, 25 patients underwent multi-vessel OPCAB surgery. Right heart support was carried out using the Enabler circulatory support system (Hemodynamics Systems Ltd., Upper Yokneam, Israel). Hemodynamic measurements were recorded at baseline and during heart displacement with and without right heart support. RESULTS: No mortality was recorded. Mean graft number was 2.25 +/- 0.36. Inability to position the Enabler system occurred in five cases. Once the Enabler was properly positioned, there was no case of conversion to cardiopulmonary bypass (CPB) due to failure of the Enabler to provide adequate support. Cardiac index (CI) declined from 2.4 +/- 0.5 L/min x /m2 to 1.6 +/- 0.6 L/min x /m2 (p <0.05) during heart displacement and increased to 2.3 +/- 0.6 L/min x /m2 (p <0.05) following Enabler activation. Mean arterial blood pressure (MAP) dropped from 82 +/- 18.6 mmHg to 53.1 +/- 16.6 mmHg (p <0.001) during heart displacement and was restored to 69.8 +/- 19.4 mmHg (p <0.001) with Enabler support. CONCLUSIONS: Heart displacement caused a significant hemodynamic deterioration that was stabilized with right heart support. The Enabler system is safe and effective in facilitating multi-vessel OPCAB surgery. However, there are still serious technical problems during cannula insertion that cause an unacceptable failure rate of device utilization. PMID- 11937456 TI - Coronary artery bypass grafting through complete sternotomy in conscious patients. AB - BACKGROUND: Complete sternotomy is the standard approach in cardiac surgery, and coronary artery bypass grafting (CABG) is the most common revascularization procedure to use this approach. A new technique has been developed that permits complete surgical revascularization with arterial grafts in awake patients. This technique, which we refer to as awake coronary artery bypass grafting, avoids general anesthesia, mechanical ventilation, and extracorporeal circulation, thereby creating the least invasive method for heart operations yet described. METHODS: A thoracic epidural catheter was placed at T2-T3 level one day before surgery. In 12 patients, single (n = 6), double (n = 5), or triple (n = 1) vessel coronary artery bypass grafting was performed without general anesthesia. In six patients, this procedure was performed after complete median sternotomy. RESULTS: Ten patients remained awake throughout the whole procedure. Two patients required secondary intubation due to incomplete analgesia in one case and pneumothorax in the other. Procedural time was 98.2 +/- 19.8 minutes. Intermediate care stay was 4.9 +/- 0.6 hours. There were no perioperative complications, and early angiographic results before discharge were excellent in all patients. CONCLUSIONS: The data presented show the feasibility and safety of complete surgical revascularization via median sternotomy using arterial grafts without general anesthesia. This approach shortens recovery time and increases patient comfort, suggesting that outpatient cardiac surgery may eventually be possible. PMID- 11937457 TI - The cardiopulmonary effects of single-lung ventilation and carbon dioxide insufflation during thoracoscopic internal mammary artery harvesting. AB - BACKGROUND: Thoracoscopic internal mammary artery harvesting has become an integral part of minimally invasive coronary artery bypass operations. The technique involves the use of single-lung ventilation and carbon dioxide insufflation to maximize exposure and facilitate rapid dissection. The hemodynamic and pulmonary effects of this technique have not been extensively studied. METHODS: Seventy-five consecutive patients undergoing a minimally invasive coronary artery bypass operation were prospectively studied intra operatively. Sixty-six left and nine right thoracoscopic IMA harvests were performed in patients with ejection fractions ranging from 12 to 70%. Carbon dioxide insufflation was utilized in the range of 8 to 12mm Hg and the effects on cardiovascular and pulmonary performance were recorded to disk every sixty seconds. RESULTS: Carbon dioxide insufflation in combination with single-lung ventilation increases central venous pressure and pulmonary artery pressure. At higher levels of insufflation pressure, negative effects are seen on systemic blood pressure, cardiac output and left ventricular stroke work. These effects can be aggravated by hypovolemia and a poor preoperative left ventricular ejection fraction. CONCLUSIONS: Single-lung ventilation and carbon dioxide insufflation greatly enhances the technical ease of thoracoscopic internal mammary artery harvest. While safe in the majority of patients, CO2 insufflation should be used cautiously in hypovolemic patients and patients with poor left ventricular function. PMID- 11937458 TI - Off-pump coronary artery surgery with the use of anastomotic devices: an additional tool for the challenging patient. AB - BACKGROUND: Avoidance of aortic side-clamping may decrease the risk of embolization to the brain and other organs during coronary revascularization. Herein we describe our preliminary experience with an anastomotic device for proximal anastomosis construction. METHODS: From October 2000 to June 2001, 18 selected patients undergoing beating heart surgery had an aorta-to-saphenous vein graft anastomosis performed with the St. Jude Medical Aortic connector. RESULTS: All anastomoses were successfully deployed. In two patients there was a minor anastomotic bleeding and in other two cases a kinking occurred due to a too short and a too long graft respectively. One patient, with graft occlusion on the fourteenth postoperative day, underwent successful percutaneous revascularization. CONCLUSIONS: Our preliminary results indicate that the aortic anastomotic device is safe and effective and its use could be widened once long term results are available. PMID- 11937459 TI - Does off-pump total arterial revascularization without aortic manipulation influence neurological outcome? A study of 226 consecutive, unselected cases. AB - BACKGROUND: Off-pump coronary artery bypass (OPCAB) is increasingly reported to have better post-operative outcomes than on-pump coronary artery bypass (ONCAB). Most series report OPCAB in selected cases and therefore the results of such studies are affected by selection bias. We report our series of consecutive, unselected OPCAB cases representing an entire coronary revascularization practice and its effect on neurological outcome. METHODS: Between January and December 2000, 226 consecutive OPCAB cases were performed. These were compared with 258 consecutive ONCAB cases performed by same surgeons historically. There were no significant differences in major demographic characteristics between the two groups. Postoperative outcomes including operation time, ventilation time, inotrope requirement, focal strokes, peak CKMB, length of hospital stay and mortality formed the basis of this study. All OPCAB patients had a median sternotomy approach. Left and right internal mammary arteries and radial arteries were used in varying combinations using composite and sequential grafting techniques to achieve revascularization and completely avoid aortic manipulation. ONCAB cases were performed employing conventional cardiopulmonary bypass techniques and using the left internal mammary artery and saphenous vein grafts as appropriate. RESULTS: OPCAB cases had significantly lower peak CKMB levels, operative time, length of hospital stay and number of grafts. There was a significantly lower incidence of permanent focal neurological events in OPCAB patients (0.4%, n = 1) compared to the ONCAB group (3.9%, n = 10, p = 0.012). CONCLUSION: This study documents the safety of off pump total arterial revascularization without aortic manipulation as the routine technique for coronary bypass surgery. It also demonstrates a significant reduction in the incidence of focal strokes. We conclude that avoidance of both cardiopulmonary bypass and aortic manipulation are important factors in reducing the occurrence of neurological deficits. PMID- 11937460 TI - Results of prospective randomized controlled trials of transmyocardial laser revascularization. AB - Over 6,000 patients worldwide have undergone transmyocardial laser revascularization (TMR) for the treatment of myocardial ischemia due to end-stage coronary artery disease since 1990. Four prospective randomized controlled trials have reported their results in comparing TMR to maximum medical therapy. All of the trials demonstrated that TMR provided significant relief of angina when compared to medical management. Additional objective data in the form of exercise tolerance and myocardial perfusion scanning was used to support the symptomatic improvement. Recent reports of the failure of percutaneous transmyocardial laser revascularization (PMR) to provide angina relief greater than that seen in a placebo group underscore the need for better understanding of TMR. While all of these trials are similar, they are not identical and this review provides an update and comparison of the results. PMID- 11937461 TI - Off-pump coronary artery bypass grafting in patients with left ventricular dysfunction. AB - BACKGROUND: Coronary artery bypass grafting in patients with severe left ventricular (LV) dysfunction still remains a high risk procedure due to its high mortality and morbidity. Off-pump surgery can be an alternative technique in these patients. We analyzed our results of off-pump coronary surgery in patients with left ventricular dysfunction and compared them with patients operated on pump. METHODS: Between January 1997 and December 2000, 355 patients with LV dysfunction (EF equal to or less than 30%) underwent off-pump coronary artery bypass (OPCAB) surgery. During the same period, 959 patients with LV dysfunction underwent coronary artery surgery on cardiopulmonary bypass. Octopus was used as mechanical stabilizer and intracoronary shunts were used in most patients. The mean age of the patients was 57.7 +/- 9.2 in patients operated on-pump and 58.4 +/- 9.8 in patients operated off-pump. RESULTS: The preoperative variables were comparable in two groups, except that there were more patients with triple vessel disease in on-pump group. Average number of grafts was 2.8 +/- 0.7 and 3.3 +/- 0.7 (p<0.001) in off-pump and on-pump groups respectively. The mortality was 3.9% and 6.0% (p = 0.176) in off-pump and on-pump groups respectively. Postoperative morbidity was less in off-pump group but it was statistically significant in incidence of atrial fibrillation and prolonged ventilation which were low in off pump group. The hospital stay was significantly less in patients operated off pump. CONCLUSION: OPCAB surgery can be safely performed in patients with LV dysfunction. The postoperative morbidity and length of stay is less as compared to patients operated on-pump. PMID- 11937462 TI - Glove retractor for left circumflex coronary artery bypass grafting. AB - BACKGROUND: During off-pump coronary artery bypass grafting (OPCAB), displacing the heart to expose the left circumflex artery (LCX) results in hemodynamic disturbance. The objective of this study was to evaluate, in a canine model, the hemodynamic impact on the beating heart of using a glove retractor instead of conventional retraction to expose the LCX. METHODS: Six mongrel dogs ranging in weight from 19.7 to 25 kg were used. Hemodynamic parameters were continuously monitored at a fixed rate of 80 beats/min. After the baseline data had been obtained, the LCX was exposed by applying an Octopus system. Each dog was then placed in the Trendelenburg position. Subsequently, the glove retractor was applied and its effects were examined both with and without the Trendelenburg position. RESULTS: LCX exposure decreased aortic flow to 35.2 +/- 12.8% of the baseline value (p< 0.001 vs. baseline), and this decrease was not fully reversed even by the Trendelenburg position (67.6 +/- 14.3%). Glove retractors tend not to interfere with right ventricular expansion so that hemodynamic disturbance is mild, and in the Trendelenburg position aortic flow (88.9 +/- 9.9%) and mean aortic pressure (95.0 +/- 5.1%) during LCX exposure were completely maintained with the glove retractor in place. CONCLUSION: Glove retractors can be used to displace the canine heart to expose the LCX, minimizing hemodynamic impairment, with normalization of hemodynamic parameters by the Trendelenburg position. This technique may offer an alternative hemodynamic support method in some patients undergoing OPCAB. PMID- 11937463 TI - Minimally invasive mitral valve replacement and multivessel coronary artery bypass through a limited right lateral thoracotomy using a balloon aortic cannula. AB - Mitral valve replacement and coronary artery bypass grafting were performed in an 80-year-old woman through an 8 cm lateral thoracotomy using central cannulation with a balloon aortic cannula. Visualization permitted the delivery of both retrograde and antegrade cardioplegia. This technique provides excellent exposure of the mitral valve and all coronary vessels and decreases the operative risks associated with a sternotomy. PMID- 11937464 TI - Video-assisted thoracoscopic surgical (VATS) closure of Patent Ductus Arteriosus: report of three-hundred cases. AB - BACKGROUND: Video-Assisted Thoracoscopic Surgery (VATS) has recently been used for Patent Ductus Arteriosus (PDA) closure on a routine basis. Our experience with this technique is supporting its efficacy. The results and advantages of VATS are the same as when thoracotomy is being performed. METHODS: From June 1997 to October 2000, there were 300 consecutive patients (mean age: 6 years old) with PDA recognized by echocardiography and/or cardiac catheterization, on whom VATS were studied. With the patients under general anesthesia, three 5 mm holes were made through the left thoracic wall. A video camera and specialized surgical devices were introduced. The ductus was dissected, and two titanium clips were applied for complete closure of the ductus. Exclusion criteria were: 1) Diameter of the ductus > 9 mm, 2) Complicated PDA such as aneurysm formation, endocarditis, and calcification, 3) Pleural adhesion and/or left sided thoracic operation in the past. RESULTS: All cases were re-assessed immediately after the procedure, and followed up by control echocardiography. No significant complication and residual shunt was recorded during the observation period. The procedure was changed to thoracotomy in three adult patients due to an inappropriately dilated canal (>9mm). Moreover, two other patients developed transient recurrent laryngeal nerve dysfunction. Mean procedure time was about 20 +/- 2 minutes. All patients were discharged shortly after the procedure (approximately 20 hours). CONCLUSION: Our results suggest that VATS compared to other techniques for PDA closure is rapid, cost-effective, painless and more cosmetic. PMID- 11937465 TI - Cardiac operations for patients with chronic liver disease. AB - BACKGROUND: Due to the systemic and hepatic effects of cardiopulmonary bypass (CPB), open-heart surgery for patients with chronic liver disease is associated with high mortality and morbidity. In this retrospective study, we present our results of cardiac surgery on patients with non-cardiac cirrhosis. METHODS: Between March 1996 and April 2000, 10 patients with chronic liver disease had open-heart surgery in our institution. Six patients were male and four were female, with a mean age of 57.1 +/- 6.85 years. Preoperative severity of liver disease was determined according to Child classification. Four cases (40%) were Child class A and six (60%) were class B. Coronary artery bypass grafting was performed in four cases, and the remaining six operations were for aortic valve replacement (AVR) and/or mitral valve replacement (MVR). Eight of the operations (80%) were performed by using cardiopulmonary bypass and two (20%) were performed as beating heart surgery. RESULTS: Chest tube drainage and transfusion needs of these patients were three times the average normal values. Three of the patients for whom CPB was used, all of them in Child class B, died. None of the patients in Child class A died. This resulted in an overall mortality rate of 30%, with mortality of 50% for the Child B group. There was no mortality for any patient who underwent cardiac surgery on the beating heart or cardiac surgery of short duration on CPB. Common characteristics of cases that were associated with high morbidity and mortality included increased postoperative hemorrhagic chest tube output, dependency on mechanical ventilation, hepatic and renal failure, gastrointestinal bleeding, and sepsis. None of the patients died of cardiac failure. CONCLUSIONS: Our findings indicate that cardiac operations may be performed with good results for patients suffering from liver disease of mild severity (Child A), but cardiac interventions that include CPB in conjunction with advanced hepatic pathologies are associated with high mortality and morbidity. Cardiac surgery (whether valvular or coronary artery surgery) for patients with chronic liver disease should be carried out with a short duration of CPB or should be done on the beating heart, if possible, in the case of coronary artery surgery. PMID- 11937466 TI - Cardiac surgical procedures and glove reinforcements. AB - The purpose of this prospective study is to determine the frequency and site of glove perforation during cardiac surgical procedures. Over a period of six months, gloves from 206 surgical team members were collected at the conclusion of surgery. All cases of known perforations were eliminated from the study. The percentage of glove perforation was 14%. The distribution of perforation across locations of the hand was significantly unequal (P = 0.001). We found that 73% of the punctures occurred in one of four contiguous locations on the glove: the radial side of the index finger (28%), the radial side of the thumb (21%), the palmar side of the index (14%), and the palmar side of the thumb (10%). Therefore, we recommend glove reinforcement on these locations that would provide better protection against transmission of infectious agents. Discomfort from restricted dexterity and impaired sense of touch with double gloving renders glove reinforcement a suitable alternative for universal precautions, especially in cardiac surgery while high level of perfection and dexterity were needed in lengthy, critical operations. PMID- 11937467 TI - Sternum screw: analysis of a novel approach to the closure of the chest after surgery. AB - BACKGROUND: To show the benefits of using a novel approach to closure of the median sternotomy through a mechanical model and mechanical testing. Simple cannulated screws are placed on either side of the sternotomy. Conventional stainless steel wire is passed through the cannula of each screw and the sternotomy is closed in the usual manner. METHODS: Hertzian contact analysis was used to estimate the stress between the wire and the sternum. Mechanical testing was used to compare using wire on its own with a sternum screw plus wire. Ten samples of balsa wood (sternum substitute) had wire placed through a hole in them, while a further ten samples were fitted with a cannulated screw and had wire passed through the screw cannula. The wire was connected to a materials testing machine, which applied tension to the wire until the wire or screw cut through the wood. RESULTS: The analysis showed that the mean stress between the wire and the sternum decreases with increasing wire diameter. At low diameters of wire the stress in the sternum can be comparable to the failure stress of bone. Using a cannulated screw reduces the stresses in the sternum. The mechanical testing showed that the wire cut through the wood at a mean load of 104 N, whereas the sternum screw cut through the wood at a mean load of 209 N (p = 0.007, Mann-Whitney Test). CONCLUSIONS: Closing a median sternotomy with cannulated screws plus wire should reduce the occurrence of sternal dehiscence. PMID- 11937468 TI - Cardioscope-assisted excision of a left ventricular tumor--a case report. AB - BACKGROUND: A 56-year-old male with a past history of excision of a malignant melanoma from his lip presented with squeezing chest pain. The patient was evaluated and determined to have a tumor of the left ventricle. Excision of such a tumor is indicated to prevent embolization and also to prevent the possibility of outflow tract obstruction. METHODS: The patient underwent transesophageal echocardiography and was placed on cardiopulmonary bypass with cold-blood cardioplegia. The ascending aorta was opened and a video-assisted cardioscope was inserted through the aortic valve and used to visualize the tumor. The tumor was resected under direct vision and the aorta was closed. RESULTS: Frozen section analysis revealed the tumor to be a benign hemangioma. The patient had an uneventful recovery with no evidence of ventricular septal defect or embolization. CONCLUSIONS: Performing the cardioscopy via the aortotomy helped us to avoid an atriotomy and/or ventriculotomy and enabled us to discern the precise extent of the tumor and rule out concomitant pathology. The use of cardioscopy as an adjunct for excision of intraventricular abnormalities can assist in determining the precise location and size of tumors and in resecting tumors in areas of the heart that might otherwise be difficult to visualize. PMID- 11937469 TI - Inferior vena caval filters or anticoagulation for patients with haemorrhagic stroke complicated by venous thromboembolism? PMID- 11937470 TI - Iron deficiency anaemia in older people: investigation, management and treatment. PMID- 11937471 TI - National initiatives in ageing research in the United Kingdom. PMID- 11937472 TI - Clinical and research challenges of intermediate care. PMID- 11937473 TI - Occult cancer in older patients presenting with venous thromboembolism. PMID- 11937474 TI - Explosive power and asymmetry in leg muscle function in frequent fallers and non fallers aged over 65. AB - BACKGROUND: although low strength is a risk factor for falls, lower limb explosive power is more predictive of functional difficulties than strength. Power may be more predictive of a future fall than strength per se. OBJECTIVE: to compare leg muscle strength and explosive power and asymmetry of leg strength and power of women aged 65 or over living at home, with and without a history of falls. DESIGN: a case controlled study of self-reported 'fallers' versus 'non fallers'. SUBJECTS: twenty women, aged 65 or over, with a history of at least three falls in the previous year were age matched with 15 women with no history of falls in the previous year. METHODS: lower limb explosive power, isometric strength of the quadriceps and hamstrings, isokinetic concentric strength (100 degrees/sec) of the quadriceps, hamstrings, ankle plantar- and dorsi-flexors and quadriceps eccentric strength (100 degrees/sec). Habitual physical activity was assessed using the self-completed Habitual Activity Profile Questionnaire. RESULTS: the women with a history of falls were less active but were not significantly weaker in any of the strength measurements, apart from ankle dorsiflexion adjusted for body weight. Both groups had significant asymmetry in all the leg muscles for both strength and power. Although both groups were asymmetrical in their lower limb power, the fallers demonstrated a significantly greater asymmetry. When the least powerful legs were compared, the women with a history of falls were 24% less powerful for their weight than those who did not fall (P=0.04). CONCLUSIONS: weakness and asymmetry is prevalent in women aged 65 and over, with and without a history of falls. Poor lower limb explosive power combined with asymmetry between limbs may be more predictive of future falls than more traditional measurements of strength in older women who live independently. PMID- 11937475 TI - Prospective randomized double-blind study of the wound-debriding effects of collagenase and fibrinolysin/deoxyribonuclease in pressure ulcers. AB - BACKGROUND: proteolytic enzymes such as collagenase, fibrinolysin and deoxyribonuclease are used for debriding purulent or fibrinous pressure ulcers. OBJECTIVE: to test the hypothesis that collagenase debrides necrotic pressure ulcers more effectively than fibrinolysin/deoxyribonuclease. METHODS: we enrolled 135 elderly patients with pressure ulcers in a randomized, prospective double blind trial. Patients were treated until complete wound debridement or for a maximum of 4 weeks with twice-daily applications of collagenase or fibrinolysin/deoxyribonuclease. The primary endpoint was percentage change in the yellow or black wound surface. Secondary endpoints were wound environment, margins, depth, pocketing, area and healing. Assessment was by two independent dermatologists who were unaware of the treatment administered and evaluated results from photographs taken at the beginning and end of treatment. RESULTS: on intention-to-treat analysis, collagenase gave slightly better results with regard to the primary endpoint in the 121 assessable patients, but this difference was not statistically significant (P=0.115). Additional efficacy measures did not show any statistically significant difference between the groups. CONCLUSION: there was no evidence of a difference between collagenase and fibrinolysin/deoxyribonuclease in the debridement of pressure ulcers. PMID- 11937476 TI - Short-term heart rate variability during a cognitive challenge in young and older adults. AB - BACKGROUND: attention-demanding tasks cause changes in the autonomic modulation of cardiac function. Heart rate variability, an index of autonomic modulation of heart rate, decreases with age. OBJECTIVE: to examine heart rate variability in elderly and young participants at rest and during an attention-demanding task. METHODS: we assessed 16 old participants (ages 72-91) and 16 college-age (ages 20 25) participants for short-term (5 min) heart rate variability at rest and during a simple-reaction time task. We report heart rate variability as the standard deviation of all interbeat intervals, and as the relative contribution of changes occurring at low- and high-frequencies. RESULTS: there were no group differences in resting heart rate. A 2x2 mixed model ANOVA suggested a main effect of age on standard deviation of all interbeat intervals (P<0.05) which was significantly lower for the older group than their younger counterparts. There was also a significant effect of the test condition on standard deviation of all interbeat intervals and spectral measures of heart rate variability (P<0.05) in that standard deviation of all interbeat intervals dropped during the simple reaction time as did high-frequencies, while normalized low frequency power increased. CONCLUSION: cardiac autonomic modulation during provocative stress show similar physiologic responses in young and older adults. PMID- 11937477 TI - Mortality predictors in disabling chronic obstructive pulmonary disease in old age. AB - OBJECTIVE: prospectively to evaluate predictors of mortality in elderly patients with disabling chronic obstructive pulmonary disease. METHODS: 137 (69 men) outpatients, aged 60-89 (mean 73) years with symptomatic disabling chronic obstructive pulmonary disease. We collected baseline demographic and physiological data. Subjects completed the Manchester Respiratory Activities of Daily Living Questionnaire, the Brief Assessment Schedule Depression Cards a screening questionnaire for depression, the Breathing Problems Questionnaire measuring quality of life, and the Montgomery Asberg Depression Rating Scale measuring severity of depression. All subjects were followed prospectively and survival and mortality data were confirmed by contacting general practitioners and scrutinising hospital notes at 30 months. RESULTS: the mean (standard deviation) of one second forced expiratory volume was 0.89 (0.3) litres. At 30 months, 44 patients (21 men, aged 61-89 [mean 75] years: 32% of the total) had died. Mean (standard deviation) baseline one second forced expiratory volume of those dying was 0.71 (0.2) litres. On logistic regression analysis, predictors of mortality were: Manchester Respiratory Activities Of Daily Living Questionnaire score (odds ratio=0.88, 95% confidence interval=0.80-0.97); pre-bronchodilator one second forced expiratory volume (odds ratio=0.04, confidence interval=0.005 0.32); body mass index (odds ratio=0.87, confidence interval=0.79-0.97); and long term oxygen therapy (odds ratio=3.17, confidence interval=1.04-8.36). Current smoking status, pack-years smoked, depression scores, quality of life scores, co morbid diseases and social class did not predict mortality. CONCLUSION: disability, use of long-term oxygen therapy, pre-bronchodilator lung function and body-mass index were independent predictors of mortality in elderly patients with severe chronic obstructive pulmonary disease. PMID- 11937478 TI - Unplanned readmission to hospital: a comparison of the views of general practitioners and hospital staff. AB - OBJECTIVE: to compare the views of general practitioners and hospital staff on the reasons for unplanned readmission of older people. METHODS: we studied 124 patients aged 65 years or over who were readmitted within 28 days of discharge. We determined the views of hospital staff and the patient's general practitioner on the reasons for readmission and compared them using McNemar's test. RESULTS: the crude readmission rate was 13.2%. The commonest agreed reason for readmission was a relapse or complication of the initial illness. Opinions differed most significantly when the reason was poor health or inadequate preparation on discharge. CONCLUSION: hospital discharge policies should take into account general practitioners' views on the causes of unplanned readmission. PMID- 11937479 TI - Caval filter placement for pulmonary embolism in a patient with a deep vein thrombosis and primary intracerebral haemorrhage. AB - Treatment of deep vein thrombosis and pulmonary embolic disease when full anticoagulation is absolutely or relatively contraindicated is difficult. We report on a case where the chosen treatment proved unsuccessful. Our patient had a primary intracerebral haemorrhage, complicated by a proximal deep vein thrombosis and despite placement of a vena caval filter, had multiple pulmonary emboli and died. PMID- 11937480 TI - Confidentiality and cognitive impairment: professional and philosophical ethics. AB - In the UK, the General Medical Council's new guidance on confidentiality restates the professional duty for doctors to regard personal information revealed to them as confidential. Such information can be shared only with the patient's explicit consent--the exceptions to this are narrowly defined. We believe the guidance does not adequately address the reality of confidentiality in modern practice, particularly in relation to patients with dementia. It seems to be naive or lacking in the subtlety required in complex clinical situations, perhaps because its legalistic, professional ethics have a philosophically limited view of confidentiality. A more sophisticated philosophical picture regards people as embedded in a shared, worldly context, in which relationships and mutual engagement become crucial. Attending to the reality of cognitively impaired people emphasizes this context and suggests that confidentiality cannot be an overriding principle--it is best regarded as a token of trust. PMID- 11937481 TI - Alternatives to hospital care. PMID- 11937482 TI - Irreversible nephrotoxicity from demeclocycline in the treatment of hyponatremia. PMID- 11937483 TI - Telecardiology: a new support for general practitioners in the management of elderly patients. PMID- 11937484 TI - Getting the right dose of repression. PMID- 11937485 TI - The rediscovery of the lymphatic system: old and new insights into the development and biological function of the lymphatic vasculature. PMID- 11937486 TI - Differentiation of embryonic stem cells is induced by GATA factors. AB - Extraembryonic endoderm (ExE) is differentiated from the inner cell mass of the late blastocyst-stage embryo to form visceral and parietal endoderm, both of which have an important role in early embryogenesis. The essential roles of Gata 6 and Gata-4 on differentiation of visceral endoderm have been identified by analyses of knockout mice. Here we report that forced expression of either Gata-6 or Gata-4 in embryonic stem (ES) cells is sufficient to induce the proper differentiation program towards ExE. We believe that this is the first report of a physiological differentiation event induced by the ectopic expression of a transcription factor in ES cells. PMID- 11937487 TI - Involvement of small RNAs and role of the qde genes in the gene silencing pathway in Neurospora. AB - Small RNA molecules have been found to be specifically associated with posttranscriptional gene silencing (PTGS) in both plants and animals. Here, we find that small sense and antisense RNAs are also involved in PTGS in Neurospora crassa. The accumulation of these RNA molecules depends on the presence of functional qde-1 and qde-3 genes previously shown to be essential for gene silencing, but does not depend on a functional qde-2, indicating that this gene is involved in a downstream step of the gene silencing pathway. Supporting this idea, a purified QDE2 protein complex was found to contain small RNA molecules, suggesting that QDE2 could be part of a small RNA-directed ribonuclease complex involved in sequence-specific mRNA degradation. PMID- 11937488 TI - A molecular link between gene-specific and chromosome-wide transcriptional repression. AB - Gene-specific and chromosome-wide mechanisms of transcriptional regulation control development in multicellular organisms. SDC-2, the determinant of hermaphrodite fate in Caenorhabditis elegans, is a paradigm for both modes of regulation. SDC-2 represses transcription of X chromosomes to achieve dosage compensation, and it also represses the male sex-determination gene her-1 to elicit hermaphrodite differentiation. We show here that SDC-2 recruits the entire dosage compensation complex to her-1, directing this X-chromosome repression machinery to silence an individual, autosomal gene. Functional dissection of her 1 in vivo revealed DNA recognition elements required for SDC-2 binding, recruitment of the dosage compensation complex, and transcriptional repression. Elements within her-1 differed in location, sequence, and strength of repression, implying that the dosage compensation complex may regulate transcription along the X chromosome using diverse recognition elements that play distinct roles in repression. PMID- 11937489 TI - Genome-wide location and regulated recruitment of the RSC nucleosome-remodeling complex. AB - Genome-wide location analysis indicates that the yeast nucleosome-remodeling complex RSC has approximately 700 physiological targets and that the Rsc1 and Rsc2 isoforms of the complex behave indistinguishably. RSC is associated with numerous tRNA promoters, suggesting that the complex is recruited by the RNA polymerase III transcription machinery. At RNA polymerase II promoters, RSC specifically targets several gene classes, including histones, small nucleolar RNAs, the nitrogen discrimination pathway, nonfermentative carbohydrate metabolism, and mitochondrial function. At the histone HTA1/HTB1 promoter, RSC recruitment requires the Hir1 and Hir2 corepressors, and it is associated with transcriptional inactivity. In contrast, RSC binds to promoters involved in carbohydrate metabolism in response to transcriptional activation, but prior to association of the Pol II machinery. Therefore, the RSC complex is generally recruited to Pol III promoters and it is specifically recruited to Pol II promoters by transcriptional activators and repressors. PMID- 11937490 TI - The role of a Williams-Beuren syndrome-associated helix-loop-helix domain containing transcription factor in activin/nodal signaling. AB - We investigated the regulation of the activin/nodal-inducible distal element (DE) of the Xenopus goosecoid (gsc) promoter. On the basis of its interaction with the DE, we isolated a Xenopus homolog of the human Williams-Beuren syndrome critical region 11 (XWBSCR11), and further, show that it interacts with pathway-specific Smad2 and Smad3 in a ligand-dependent manner. Interestingly, we also find that XWBSCR11 functions cooperatively with FoxH1 (Fast-1) to stimulate DE-dependent transcription. We propose a mechanism in which FoxH1 functions together with Smads as a cofactor for the recruitment of transcription factors like XWBSCR11 in the process of activin/nodal-mediated gsc-specific induction. This mechanism provides considerable opportunities for modulation of transcription across a variety of activin/nodal-inducible genes, increasing diversity in promoter selection, thus leading to the differential induction of activin/nodal target genes. PMID- 11937491 TI - The plexin-B1/Rac interaction inhibits PAK activation and enhances Sema4D ligand binding. AB - The small GTPase Rac has been implicated in growth cone guidance mediated by semaphorins and their receptors. Here we demonstrate that plexin-B1, a receptor for Semaphorin4D (Sema4D), and p21-activated kinase (PAK) can compete for the interaction with active Rac and plexin-B1 can inhibit Rac-induced PAK activation. We have also demonstrated that expression of active Rac enhances the ability of plexin-B1 to interact with Sema4D. Active Rac stimulates the localization of plexin-B1 to the cell surface. The enhancement in Sema4D binding depends on the ability of Rac to bind plexin-B1. These observations support a model where signaling between Rac and plexin-B1 is bidirectional; Rac modulates plexin-B1 activity and plexin-B1 modulates Rac function. PMID- 11937492 TI - Notch pathway molecules are essential for the maintenance, but not the generation, of mammalian neural stem cells. AB - Neural stem cells, which exhibit self-renewal and multipotentiality, are generated in early embryonic brains and maintained throughout the lifespan. The mechanisms of their generation and maintenance are largely unknown. Here, we show that neural stem cells are generated independent of RBP-Jkappa, a key molecule in Notch signaling, by using RBP-Jkappa(-/-) embryonic stem cells in an embryonic stem cell-derived neurosphere assay. However, Notch pathway molecules are essential for the maintenance of neural stem cells; they are depleted in the early embryonic brains of RBP-Jkappa(-/-) or Notch1(-/-) mice. Neural stem cells also are depleted in embryonic brains deficient for the presenilin1 (PS1) gene, a key regulator in Notch signaling, and are reduced in PS1(+/-) adult brains. Both neuronal and glial differentiation in vitro were enhanced by attenuation of Notch signaling and suppressed by expressing an active form of Notch1. These data are consistent with a role for Notch signaling in the maintenance of the neural stem cell, and inconsistent with a role in a neuronal/glial fate switch. PMID- 11937493 TI - Coordination of chondrogenesis and osteogenesis by fibroblast growth factor 18. AB - Gain of function mutations in fibroblast growth factor (FGF) receptors cause chondrodysplasia and craniosynostosis syndromes. The ligands interacting with FGF receptors (FGFRs) in developing bone have remained elusive, and the mechanisms by which FGF signaling regulates endochondral, periosteal, and intramembranous bone growth are not known. Here we show that Fgf18 is expressed in the perichondrium and that mice homozygous for a targeted disruption of Fgf18 exhibit a growth plate phenotype similar to that observed in mice lacking Fgfr3 and an ossification defect at sites that express Fgfr2. Mice lacking either Fgf18 or Fgfr3 exhibited expanded zones of proliferating and hypertrophic chondrocytes and increased chondrocyte proliferation, differentiation, and Indian hedgehog signaling. These data suggest that FGF18 acts as a physiological ligand for FGFR3. In addition, mice lacking Fgf18 display delayed ossification and decreased expression of osteogenic markers, phenotypes not seen in mice lacking Fgfr3. These data demonstrate that FGF18 signals through another FGFR to regulate osteoblast growth. Signaling to multiple FGFRs positions FGF18 to coordinate chondrogenesis in the growth plate with osteogenesis in cortical and trabecular bone. PMID- 11937494 TI - FGF18 is required for normal cell proliferation and differentiation during osteogenesis and chondrogenesis. AB - Fibroblast growth factor (FGF) signaling is involved in skeletal development of the vertebrate. Gain-of-function mutations of FGF receptors (FGFR) cause craniosynostosis, premature fusion of the skull, and dwarfism syndromes. Disruption of Fgfr3 results in prolonged growth of long bones and vertebrae. However, the role that FGFs actually play in skeletal development in the embryo remains unclear. Here we show that Fgf18 is expressed in and required for osteogenesis and chondrogenesis in the mouse embryo. Fgf18 is expressed in both osteogenic mesenchymal cells and differentiating osteoblasts during calvarial bone development. In addition, Fgf18 is expressed in the perichondrium and joints of developing long bones. In calvarial bone development of Fgf18-deficient mice generated by gene targeting, the progress of suture closure is delayed. Furthermore, proliferation of calvarial osteogenic mesenchymal cells is decreased, and terminal differentiation to calvarial osteoblasts is specifically delayed. Delay of osteogenic differentiation is also observed in the developing long bones of this mutant. Conversely, chondrocyte proliferation and the number of differentiated chondrocytes are increased. Therefore, FGF18 appears to regulate cell proliferation and differentiation positively in osteogenesis and negatively in chondrogenesis. PMID- 11937495 TI - Role of DIVARICATA in the control of dorsoventral asymmetry in Antirrhinum flowers. AB - Dorsoventral asymmetry of the Antirrhinum corolla depends on expression of the CYC and DICH genes in dorsal petals. One role of these genes is to inhibit DIVARICATA (DIV), a determinant of ventral identity. Therefore, in cyc;dich double mutants ventral identity spreads all around the flower. We show that DIV encodes a protein belonging to the MYB family of transcription factors. Early on in corolla development, DIV affects specifically the growth of ventral and lateral petals but is transcribed in all petals. Analysis of a closely related gene suggests that the lack of effect on dorsal petals is not due to redundancy. More likely, therefore, DIV is regulated posttranscriptionally through a mechanism that depends on CYC and DICH. Later on, DIV affects growth and cell types and is transcribed mostly in a single layer of cells of ventral and lateral petals. This late pattern may itself depend on DIV activity because it fails to be established in a transcribed but inactive div mutant and, conversely, spreads all around the flower in cyc;dich double mutants. PMID- 11937496 TI - Select HIV protease inhibitors alter bone and fat metabolism ex vivo. AB - Human immunodeficiency virus (HIV) therapies have been associated with alterations in fat metabolism and bone mineral density. This study examined the effects of HIV protease inhibitors (PIs) on bone resorption, bone formation, and adipocyte differentiation using ex vivo cultured osteoclasts, osteoblasts, and adipocytes, respectively. Osteoclast activity, measured using a rat neonatal calvaria assay, increased in the presence of nelfinavir (NFV; 47.2%, p = 0.001), indinavir (34.6%, p = 0.001), saquinavir (24.3%, p = 0.001), or ritonavir (18%, p < 0.01). In contrast, lopinavir (LPV) and amprenavir did not increase osteoclast activity. In human mesenchymal stem cells (hMSCs), the PIs LPV and NFV decreased osteoblast alkaline phosphatase enzyme activity and gene expression significantly (p < 0.05). LPV and NFV diminished calcium deposition and osteoprotegrin expression (p < 0.05), whereas the other PIs investigated did not. Adipogenesis of hMSCs was strongly inhibited by saquinavir and NFV (>50%, p < 0.001) and moderately inhibited by ritonavir and LPV (>40%, p < 0.01). Expression of diacylglycerol transferase, a marker of adipocyte differentiation, decreased in hMSCs treated with NFV. Amprenavir and indinavir did not affect adipogenesis or lipolysis. These results suggest that bone and fat formation in hMSCs of bone marrow may be coordinately down-regulated by some but not all PIs. PMID- 11937497 TI - Effects of nitroglycerin on soluble guanylate cyclase: implications for nitrate tolerance. AB - Soluble guanylate cyclase (sGC) is a heterodimeric hemoprotein that catalyzes the conversion of GTP to cGMP. Upon binding NO to its heme cofactor, purified sGC was activated 300-fold. sGC was only activated 67-fold by nitroglycerin (GTN) and Cys; and in the absence of Cys, GTN did not activate sGC. Electronic absorption spectroscopy studies showed that upon NO binding, the Soret of ferrous sGC shifted from 431 to 399 nm. The data also revealed that activation of sGC by GTN/Cys was not via the expected ferrous heme-NO species as indicated by the absence of the 399 nm heme Soret. Furthermore, EPR studies of the reaction of GTN/Cys with sGC confirmed that no ferrous heme-NO species was formed but that there was heme oxidation. Potassium ferricyanide is known to oxidize ferrous sGC to the ferric oxidation state. Spectroscopic and activity data for the reactions of sGC with GTN alone or with K(3)Fe(CN)(6) were indistinguishable. These data suggest the following: 1) GTN/Cys do not activate sGC via GTN biotransformation to NO in vitro, and 2) in the absence of added thiol, GTN oxidizes sGC. PMID- 11937498 TI - Reconstitution of hepatitis C virus envelope glycoproteins into liposomes as a surrogate model to study virus attachment. AB - The envelope glycoproteins, E1 and E2, of hepatitis C virus (HCV) assemble intracellularly to form a noncovalent heterodimer that is expected to be essential for viral assembly and entry. However, due to the lack of a cell culture system supporting efficient HCV replication, it is very difficult to obtain relevant information on the functions of this glycoprotein oligomer. To get better insights into its biological and biochemical properties, HCV envelope glycoprotein heterodimer expressed by a vaccinia virus recombinant was purified by immunoaffinity. Purified E1E2 heterodimer was recognized by conformation dependent monoclonal antibodies, showing that the proteins were properly folded. In addition, it interacted with human CD81, a putative HCV receptor, as well as with human low and very low density lipoproteins, which have been shown to be associated with infectious HCV particles isolated from patients. Purified E1E2 heterodimer was also reconstituted into liposomes. E1E2-liposomes were recognized by a conformation-dependent monoclonal antibody as well as by human CD81. Together, these data indicate that E1E2-liposomes are a valuable tool to study the molecular requirements for HCV binding to target cells. PMID- 11937499 TI - Calpain inhibition decreases the growth rate of mammalian cell colonies. AB - The calpains, a family of calcium-requiring intracellular proteases, are proposed regulators of cell proliferation. However, ablation of the calpain small subunit gene necessary for function of the conventional calpains did not result in decreased rate of proliferative growth of mouse stem cells under routine culture conditions. To address the reasons for this discrepancy, Chinese hamster ovary cell lines were established that overexpress the calpain inhibitor protein, calpastatin, under control of the ecdysone congener, ponasterone A. Overexpression of calpastatin in these cell lines resulted in a decreased growth of isolated colonies adhering to tissue culture plates. However, when cells were plated at higher density, calpastatin overexpression had no influence on proliferative growth rate. Growth of colonies in soft agar was not inhibited by calpastatin overexpression. Cell adhesion, cell de-adhesion, and cell motility all appeared to be normal after calpastatin overexpression. Differential display analysis was initiated to detect possible alteration of gene expression upon calpastatin overexpression. Analysis of approximately 3000 differential display PCR signals resulted in identification of one band that was underexpressed. Northern blot analysis confirmed a decreased amount of approximately 1 kb mRNA in cells overexpressing calpastatin. Sequence analysis identified a putative protein, Csr, containing a region homologous to two ubiquitin transferases and a putative cation channel protein. PMID- 11937500 TI - Reciprocal protein kinase A regulatory interactions between cystic fibrosis transmembrane conductance regulator and Na+/H+ exchanger isoform 3 in a renal polarized epithelial cell model. AB - Although Cystic fibrosis transmembrane conductance regulator (CFTR) has been shown to regulate the activity of NHE3, the potential reciprocal interaction of NHE3 to modulate the protein kinase A (PKA)-dependent regulation of CFTR in epithelial cells is still unknown. In the present work, we describe experiments to define the interactions between CFTR and NHE3 with the regulatory, scaffolding protein, NHERF that organize their PKA-dependent regulation in a renal epithelial cell line that expresses endogenous CFTR. The expression of rat NHE3 significantly decreased PKA-dependent activation of CFTR without altering CFTR expression, and this decrease was prevented by mutation of either of the two rat NHE3 PKA target serines to alanine (S552A or S605A). Inhibition of CFTR expression by antisense treatment resulted in an acute decrease in PKA-dependent regulation of NHE3 activity. CFTR, NHE3, and ezrin were recognized by NHERF-2 but not NHERF-1 in glutathione S-transferase pull-down experiments. Ezrin may function as a protein kinase A anchoring protein (AKAP) in this signaling complex, because blocking the binding of PKA to an AKAP by incubation with the S Ht31 peptide inhibited the PKA-dependent regulation of CFTR in the absence of NHE3. In the A6-NHE3 cells S-Ht31 blocked the PKA regulation of NHE3 whereas it now failed to affect the regulation of CFTR. We conclude that CFTR and NHE3 reciprocally interact via a shared regulatory complex comprised of NHERF-2, ezrin, and PKA. PMID- 11937501 TI - Selectivity and promiscuity of the first and second PDZ domains of PSD-95 and synapse-associated protein 102. AB - PDZ domains typically interact with the very carboxyl terminus of their binding partners. Type 1 PDZ domains usually require valine, leucine, or isoleucine at the very COOH-terminal (P(0)) position, and serine or threonine 2 residues upstream at P(-2). We quantitatively defined the contributions of carboxyl terminal residues to binding selectivity of the prototypic interactions of the PDZ domains of postsynaptic density protein 95 (PSD-95) and its homolog synapse associated protein 90 (SAP102) with the NR2b subunit of the N-methyl-d-aspartate type glutamate receptor. Our studies indicate that all of the last five residues of NR2b contribute to the binding selectivity. Prominent were a requirement for glutamate or glutamine at P(-3) and for valine at P(0) for high affinity binding and a preference for threonine over serine at P(-2), in the context of the last 11 residues of the NR2b COOH terminus. This analysis predicts a COOH-terminal (E/Q)(S/T)XV consensus sequence for the strongest binding to the first two PDZ domains of PSD-95 and SAP102. A search of the human genome sequences for proteins with a COOH-terminal (E/Q)(S/T)XV motif yielded 50 proteins, many of which have not been previously identified as PSD-95 or SAP102 binding partners. Two of these proteins, brain-specific angiogenesis inhibitor 1 and protein kinase Calpha, co immunoprecipitated with PSD-95 and SAP102 from rat brain extracts. PMID- 11937502 TI - Structural characterizations of fusion peptide analogs of influenza virus hemagglutinin. Implication of the necessity of a helix-hinge-helix motif in fusion activity. AB - Infection by enveloped viruses initially involves membrane fusion between viral and host cell membranes. The fusion peptide plays a crucial role in triggering this reaction. To clarify how the fusion peptide exerts this specific function, we carried out biophysical studies of three fusion peptide analogs of influenza virus hemagglutinin HA2, namely E5, G13L, and L17A. E5 exhibits an activity similar to the native fusion peptide, whereas G13L and L17A, which are two point mutants of the E5 analog, possess much less fusion activity. Our CD data showed that the conformations of these three analogs in SDS micelles are pH-dependent, with higher alpha-helical contents at acidic pH. Tryptophan fluorescence emission experiments indicated that these three analogs insert deeper into lipid bilayers at acidic pH. The three-dimensional structure of the E5 analog in SDS micelles at pH 4.0 revealed that two segments, Leu(2)-Glu(11) and Trp(14)-Ile(18), form amphipathic helical conformations, with Gly(12)-Gly(13) forming a hinge. The hydrophobic residues in the N- and C-terminal helices form a hydrophobic cluster. At neutral pH, however, the C-terminal helix of Trp(14)-Ile(18) reduces dramatically, and the hydrophobic core observed at acidic pH is severely disrupted. We suggest that the disruption of the C-terminal helix renders the E5 analog fusion-inactive at neutral pH. Furthermore, the decrease of the hinge and the reduction of fusion activity in G13L reveal the importance of the hinge in fusion activity. Also, the decrease in the C-terminal helix and the reduction of fusion activity in L17A demonstrates the importance of the C-terminal helix in fusion activity. Based on these biophysical studies, we propose a model that illustrates the structural change of the HA2 fusion peptide analog and explains how the analog interacts with the lipid bilayer at different pH values. PMID- 11937503 TI - A novel zinc-regulated human zinc transporter, hZTL1, is localized to the enterocyte apical membrane. AB - Zinc is essential to a wide range of cellular processes; therefore, it is important to elucidate the molecular mechanisms of zinc homeostasis. To date, no zinc transporters expressed at the enterocyte apical membrane, and so essential to mammalian zinc homeostasis, have been discovered. We identified hZTL1 as a human expressed sequence tag with homology to the basolateral enterocyte zinc transporter ZnT1 and deduced the full-length cDNA sequence by PCR. The protein of 523 amino acids belongs to the cation diffusion facilitator family of membrane transporters. Unusually, the predicted topology comprises 12 rather than 6 transmembrane domains. ZTL1 mRNA was detected by reverse transcription-PCR in a range of mouse tissues. A Myc-tagged hZTL1 clone was expressed in transiently transfected polarized human intestinal Caco-2 cells at the apical membrane. Expression of hZTL1 mRNA in Caco-2 cells increased with zinc supplementation of the nutrient medium; however, in the placental cell line JAR hZTL1 appeared not to be regulated by zinc. Heterologous expression of hZTL1 in Xenopus laevis oocytes increased zinc uptake across the plasma membrane. The localization, regulatory properties, and function of hZTL1 indicate a role in regulating the absorption of dietary zinc across the apical enterocyte membrane. PMID- 11937504 TI - Interaction of transcriptional intermediary factor 2 nuclear receptor box peptides with the coactivator binding site of estrogen receptor alpha. AB - The activation function 2/ligand-dependent interaction between nuclear receptors and their coregulators is mediated by a short consensus motif, the so-called nuclear receptor (NR) box. Nuclear receptors exhibit distinct preferences for such motifs depending both on the bound ligand and on the NR box sequence. To better understand the structural basis of motif recognition, we characterized the interaction between estrogen receptor alpha and the NR box regions of the p160 coactivator TIF2. We have determined the crystal structures of complexes between the ligand-binding domain of estrogen receptor alpha and 12-mer peptides from the Box B2 and Box B3 regions of TIF2. Surprisingly, the Box B3 module displays an unexpected binding mode that is distinct from the canonical LXXLL interaction observed in other ligand-binding domain/NR box crystal structures. The peptide is shifted along the coactivator binding site in such a way that the interaction motif becomes LXXYL rather than the classical LXXLL. However, analysis of the binding properties of wild type NR box peptides, as well as mutant peptides designed to probe the Box B3 orientation, suggests that the Box B3 peptide primarily adopts the "classical" LXXLL orientation in solution. These results highlight the potential difficulties in interpretation of protein-protein interactions based on co-crystal structures using short peptide motifs. PMID- 11937505 TI - The Caenorhabditis elegans gene, gly-2, can rescue the N acetylglucosaminyltransferase V mutation of Lec4 cells. AB - UDP-N-acetylglucosamine:alpha-6-d-mannoside beta-1,6-N acetylglucosaminyltransferase V (GlcNAc-TV) is a regulator of polylactosamine containing N-glycans and is causally involved in T cell regulation and tumor metastasis. The Caenorhabditis elegans genome contains a single orthologous gene, gly-2, that is transcribed and encodes a 669-residue type II membrane protein that is 36.7% identical to mammalian GlcNAc-TV (Mgat-5). Recombinant GLY-2 possessed GlcNAc-TV activity when assayed in vitro, and protein truncations demonstrated that the N-terminal boundary of the catalytic domain is Ile-138. gly 2 complemented the Phaseolus vulgaris leucoagglutinin binding defect of Chinese hamster ovary Lec4 cells, whereas GLY-2(L116R), an equivalent mutation to that which causes the Lec4A phenotype, could not. We conclude that the worm gene is functionally interchangeable with the mammalian form. GlcNAc-TV activity was detected in wild-type animals but not those homozygous for a deletion allele of gly-2. Activity was restored in mutant animals by an extrachromosomal array that encompassed the gly-2 gene. Green fluorescent protein reporter transgenes driven by the gly-2 promoter were expressed by developing embryos from the late comma stage onward, present in a complex subset of neurons in larvae and, in addition, the spermathecal and pharyngeal-intestinal valves and certain vulval cells of adults. However, no overt phenotypes were observed in animals homozygous for deletion alleles of gly-2. PMID- 11937506 TI - Multiple factors prevent transcriptional interference at the yeast ARO4-HIS7 locus. AB - Increased transcriptional activity may cause transcriptional interference in organisms with compact genomes such as the yeast Saccharomyces cerevisiae. Replacement of the yeast ARO4 promoter by the stronger ACT1 promoter increases ARO4 transcription and simultaneously reduces the basal transcription of the downstream HIS7 gene. The open reading frames of ARO4 and HIS7 are tandemly transcribed and are separated by 416 bp. In wild-type cells, a nuclease-resistant site suggests that the two genes are separated by a single positioned nucleosome. Transcriptional interference correlates with Micrococcus nuclease accessibility of this otherwise nuclease-resistant site. Deletion analyses of the region between the two open reading frames revealed that transcriptional interference increases upon removal of either parts of the ARO4 3' end or HIS7 promoter sequences. The abolishment of the Abf1p-binding site within the HIS7 promoter significantly enhances transcriptional interference, resulting in a histidine auxotrophic strain. Our data suggest that the yeast cell prevents transcriptional interference by the combined action of efficient ARO4 transcription termination, the positioning of a fixed nucleosome, and transcription factor binding to the HIS7 promoter. PMID- 11937507 TI - Species-specific and mutant MWFE proteins. Their effect on the assembly of a functional mammalian mitochondrial complex I. AB - The MWFE protein (70 amino acids) is highly conserved in evolution, but the human protein (80% identical to hamster) does not complement a null mutation in Chinese hamster cells. We have identified a small protein segment where significant differences exist between rodents and primates, illustrating very specifically the need for compatibility of the nuclear and mitochondrial genomes in the assembly of complex I. The segment between amino acids 39 and 46 appears to be critical for species-specific compatibility. Amino acid substitutions in this region were tested that caused a reduction of activity of the hamster protein or converted the inactive human protein into a partially active one. Such mutations could be useful in making mice with partial complex I activity as models for mitochondrial diseases. Their potential as dominant negative mutants was explored. More deleterious mutations in the NDUFA1 gene were also characterized. A conservative substitution, R50K, or a short C-terminal deletion makes the protein completely inactive. In the absence of MWFE, no high molecular weight complex was detectable by Blue Native-gel electrophoresis. The MWFE protein itself is unstable in the absence of assembled mitochondrially encoded integral membrane proteins of complex I. PMID- 11937508 TI - Direct interaction of dermaseptin S4 aminoheptanoyl derivative with intraerythrocytic malaria parasite leading to increased specific antiparasitic activity in culture. AB - Antiplasmodial activity of the dermaseptin S4 derivative K(4)S4(1-13) (P) was shown to be mediated by lysis of the host cells. To identify antiplasmodial peptides with enhanced selectivity, we produced and screened new derivatives based on P and singled out the aminoheptanoylated peptide (NC7-P) for its improved antiplasmodial properties. Compared with P, NC7-P displayed both increased antiparasitic efficiency and reduced hemolysis, including against infected cells. Antiplasmodial activity of P and its derivative was time dependent and irreversible, implying a cytotoxic effect. But, whereas the dose dependence of growth inhibition and hemolysis of infected cells overlapped when treated with P, NC7-P exerted more than 50% growth inhibition at peptide concentrations that did not cause hemolysis. Noticeably, NC7-P but not P, dissipated the parasite plasma membrane potential and caused depletion of intraparasite potassium at nonhemolytic conditions. Confocal microscopy analysis of infected cells localized the rhodaminated derivative in association with parasite membranes and intraerythrocytic tubulovesicular structures, whereas in normal cells, the peptide localized exclusively at the plasma membrane. Overall, the data demonstrate that antimicrobial peptides can be engineered to act specifically on the membrane of intracellular parasites and support a mechanism whereby NC7-P crosses the host cell plasma membrane and disrupts the parasite membrane(s). PMID- 11937509 TI - Scanning mutagenesis of a Janus-faced atracotoxin reveals a bipartite surface patch that is essential for neurotoxic function. AB - The Janus-faced atracotoxins (J-ACTXs) are a family of insect-specific excitatory neurotoxins isolated from the venom of Australian funnel web spiders. In addition to a strikingly asymmetric distribution of charged residues, from which their name is derived, these toxins contain an extremely rare vicinal disulfide bond. To shed light on the mechanism of action of these toxins and to enhance their utility as lead compounds for insecticide development, we developed a recombinant expression system for the prototypic family member, J-ACTX-Hv1c, and mapped the key functional residues using site-directed mutagenesis. An alanine scan using a panel of 24 mutants provided the first complete map of the bioactive surface of a spider toxin and revealed that the entire J-ACTX-Hv1c pharmacophore is restricted to seven residues that form a bipartite surface patch on one face of the toxin. However, the primary pharmacophore, or hot spot, is formed by just five residues (Arg(8), Pro(9), Tyr(31), and the Cys(13)-Cys(14) vicinal disulfide). The Arg(8) Tyr(31) diad in J-ACTX-Hv1c superimposes closely on the Lys-(Tyr/Phe) diad that is spatially conserved across a range of structurally dissimilar K(+) channel blockers, which leads us to speculate that the J-ACTXs might target an invertebrate K(+) channel. PMID- 11937510 TI - Function assignment to conserved residues in mammalian alkaline phosphatases. AB - We have probed the structural/functional relationship of key residues in human placental alkaline phosphatase (PLAP) and compared their properties with those of the corresponding residues in Escherichia coli alkaline phosphatase (ECAP). Mutations were introduced in wild-type PLAP, i.e. [E429]PLAP, and in some instances also in [G429]PLAP, which displays properties characteristic of the human germ cell alkaline phosphatase isozyme. All active site metal ligands, as well as residues in their vicinity, were substituted to alanines or to the homologous residues present in ECAP. We found that mutations at Zn2 or Mg sites had similar effects in PLAP and ECAP but that the environment of the Zn1 ion in PLAP is less affected by substitutions than that in ECAP. Substitutions of the Mg and Zn1 neighboring residues His-317 and His-153 increased k(cat) and increased K(m) when compared with wild-type PLAP, contrary to what was predicted by the reciprocal substitutions in ECAP. All mammalian alkaline phosphatases (APs) have five cysteine residues (Cys-101, Cys-121, Cys-183, Cys-467, and Cys-474) per subunit, not homologous to any of the four cysteines in ECAP. By substituting each PLAP Cys by Ser, we found that disrupting the disulfide bond between Cys-121 and Cys-183 completely prevents the formation of the active enzyme, whereas the carboxyl-terminally located Cys-467-Cys-474 bond plays a lesser structural role. The substitution of the free Cys-101 did not significantly affect the properties of the enzyme. A distinguishing feature found in all mammalian APs, but not in ECAP, is the Tyr-367 residue involved in subunit contact and located close to the active site of the opposite subunit. We studied the A367 and F367 mutants of PLAP, as well as the corresponding double mutants containing G429. The mutations led to a 2-fold decrease in k(cat), a significant decrease in heat stability, and a significant disruption of inhibition by the uncompetitive inhibitors l-Phe and l-Leu. Our mutagenesis data, computer modeling, and docking predictions indicate that this residue contributes to the formation of the hydrophobic pocket that accommodates and stabilizes the side chain of the inhibitor during uncompetitive inhibition of mammalian APs. PMID- 11937511 TI - Six related nucleoside/nucleobase transporters from Trypanosoma brucei exhibit distinct biochemical functions. AB - Purine nucleoside and nucleobase transporters are of fundamental importance for Trypanosoma brucei and related kinetoplastid parasites because these protozoa are not able to synthesize purines de novo and must salvage the compounds from their hosts. In the studies reported here, we have identified a family of six clustered genes in T. brucei that encode nucleoside/nucleobase transporters. These genes, TbNT2/927, TbNT3, TbNT4, TbNT5, TbNT6, and TbNT7, have predicted amino acid sequences that show high identity to each other and to TbNT2, a P1 type nucleoside transporter recently identified in our laboratory. Expression in Xenopus laevis oocytes revealed that TbNT2/927, TbNT5, TbNT6, and TbNT7 are high affinity adenosine/inosine transporters with K(m) values of <5 microm. In addition, TbNT5, and to a limited degree TbNT6 and TbNT7, also mediate the uptake of the nucleobase hypoxanthine. Ribonuclease protection assays showed that mRNA from all of the six members of this gene family are expressed in the bloodstream stage of the T. brucei life cycle but that TbNT2/927 and TbNT5 mRNAs are also expressed in the insect stage of the life cycle. These results demonstrate that T. brucei expresses multiple purine transporters with distinct substrate specificities and different patterns of expression during the parasite life cycle. PMID- 11937512 TI - Biosynthesis of the C(7)-cyclitol moiety of acarbose in Actinoplanes species SE50/110. 7-O-phosphorylation of the initial cyclitol precursor leads to proposal of a new biosynthetic pathway. AB - We have previously demonstrated that the biosynthesis of the C(7)-cyclitol, called valienol (or valienamine), of the alpha-glucosidase inhibitor acarbose starts from the cyclization of sedo-heptulose 7-phosphate to 2-epi-5-epi valiolone (Stratmann, A., Mahmud, T., Lee, S., Distler, J., Floss, H. G., and Piepersberg, W. (1999) J. Biol. Chem. 274, 10889-10896). Synthesis of the intermediate 2-epi-5-epi-valiolone is catalyzed by the cyclase AcbC encoded in the biosynthetic (acb) gene cluster of Actinoplanes sp. SE50/110. The acbC gene lies in a possible transcription unit, acbKLMNOC, cluster encompassing putative biosynthetic genes for cyclitol conversion. All genes were heterologously expressed in strains of Streptomyces lividans 66 strains 1326, TK23, and TK64. The AcbK protein was identified as the acarbose 7-kinase, which had been described earlier (Drepper, A., and Pape, H. (1996) J. Antibiot. (Tokyo) 49, 664 668). The multistep conversion of 2-epi-5-epi-valiolone to the final cyclitol moiety was studied by testing enzymatic mechanisms such as dehydration, reduction, epimerization, and phosphorylation. Thus, a phosphotransferase activity was identified modifying 2-epi-5-epi-valiolone by ATP-dependent phosphorylation. This activity could be attributed to the AcbM protein by verifying this activity in S. lividans strain TK64/pCW4123M, expressing His tagged AcbM. The His-tagged AcbM protein was purified and subsequently characterized as a 2-epi-5-epi-valiolone 7-kinase, presumably catalyzing the first enzyme reaction in the biosynthetic route, leading to an activated form of the intermediate 1-epi-valienol. The AcbK protein could not catalyze the same reaction nor convert any of the other C(7)-cyclitol monomers tested. The 2-epi-5 epi-valiolone 7-phosphate was further converted by the AcbO protein to another isomeric and phosphorylated intermediate, which was likely to be the 2-epimer 5 epi-valiolone 7-phosphate. The products of both enzyme reactions were characterized by mass spectrometric methods. The product of the AcbM-catalyzed reaction, 2-epi-5-epi-valiolone 7-phosphate, was purified on a preparative scale and identified by NMR spectroscopy. A biosynthetic pathway for the pseudodisaccharidic acarviosyl moiety of acarbose is proposed on the basis of these data. PMID- 11937513 TI - Salicylate biosynthesis in Pseudomonas aeruginosa. Purification and characterization of PchB, a novel bifunctional enzyme displaying isochorismate pyruvate-lyase and chorismate mutase activities. AB - Isochorismate pyruvate-lyase (IPL), the second enzyme of pyochelin biosynthesis and the product of the pchB gene, was purified to homogeneity from Pseudomonas aeruginosa. In the reaction catalyzed by this enzyme, isochorismate --> salicylate + pyruvate, no cofactors appear to be required. At the pH optimum (pH 6.8), the enzyme displayed Michaelis-Menten kinetics, with an apparent K(m) of 12.5 microm for isochorismate and a kcat of 106 min(-1), calculated per monomer. The native enzyme behaved as a homodimer, as judged by molecular sieving chromatography, electrophoresis under nondenaturing conditions, and cross-linking experiments. PchB has approximately 20% amino acid sequence identity with AroQ class chorismate mutases (CMs). Chorismate was shown to be converted to prephenate by purified PchB in vitro, with an apparent K(m) of 150 microm and a kcat of 7.8 min(-1). An oxabicyclic diacid transition state analog and well characterized inhibitor of CMs competitively inhibited both IPL and CM activities of PchB. Moreover, a CM-deficient Escherichia coli mutant, which is auxotrophic for phenylalanine and tyrosine, was functionally complemented by the cloned P. aeruginosa pchB gene for growth in minimal medium. A mutant form of PchB, in which isoleucine 88 was changed to threonine, had no detectable IPL activity, but retained wild-type CM activity. In conclusion, the 11.5-kDa subunit of PchB appears to contain a single active site involved in both IPL and CM activity. PMID- 11937514 TI - Identification and characterization of a sphingolipid delta 4-desaturase family. AB - Sphingolipids desaturated at the Delta4-position are important signaling molecules in many eukaryotic organisms, including mammals. In a bioinformatics approach, we now identified a new family of protein sequences from animals, plants, and fungi and characterized these sequences biochemically by expression in Saccharomyces cerevisiae. This resulted in the identification of the enzyme sphingolipid Delta4-desaturase (dihydroceramide desaturase) from Homo sapiens, Mus musculus, Drosophila melanogaster, and Candida albicans, in addition to a bifunctional sphingolipid Delta4-desaturase/C-4-hydroxylase from M. musculus. Among the sequences investigated are the Homo sapiens membrane lipid desaturase, the M. musculus degenerative spermatocyte, and the Drosophila melanogaster degenerative spermatocyte proteins. During spermatogenesis, but not oogenesis of des mutant flies, both cell cycle and spermatid differentiation are specifically blocked at the entry into the first meiotic division, leading to male sterility. This mutant phenotype can be restored to wild-type by complementation with a functional copy of the des gene (Endo, K., Akiyama, T., Kobayashi S., and Okada, M. (1996) Mol. Gen. Genet. 253, 157-165). These results suggest that Delta4 desaturated sphingolipids provide an early signal necessary to trigger the entry into both meiotic and spermatid differentiation pathways during Drosophila spermatogenesis. PMID- 11937515 TI - Comparative studies of the mode of oxidation of phenyl derivatives of fatty acids by the animal organism and by hydrogen peroxide (Dakin, H. D. (1908) J. Biol. Chem. 4, 419-435). PMID- 11937517 TI - In memoriam: G. Jeanette Thorbecke 1929-2001. PMID- 11937518 TI - Cutting edge: biasing immune responses by directing antigen to macrophage Fc gamma receptors. AB - An immune response can deviate toward either a Th1- or Th2-like response. In this work we examine the contribution that activated macrophages and IgG Abs make toward this deviation. The use of activated macrophages as APCs resulted in a strong polarized T cell response that was predominated by IFN-gamma. However, when Ag was targeted to FcgammaRs on these macrophages, the T cell response was reversed and biased toward a Th2-like response. This Th2-like phenotype was stable and was retained when the T cells were subsequently restimulated under nonbiasing conditions. The T cell biasing and its reversal via FcgammaR was also observed in vivo. Mice vaccinated with IgG-opsonized OVA made high levels of IgG Ab of the IgG1 isotype. These studies demonstrate that the ligation of FcgammaR on activated macrophages can reverse the Th1 biasing that occurs as a result of innate immune responses to microbial products. PMID- 11937519 TI - Cutting edge: DNA polymerases mu and lambda are dispensable for Ig gene hypermutation. AB - Mutations arising in Ig V genes during an immune response are most likely introduced by one or several error-prone DNA polymerases. Many of the recently described nonreplicative DNA polymerases have an intrinsic fidelity compatible with such an activity, the strongest candidates being polymerase (pol) eta, pol iota, pol zeta, and pol mu. We report in this work that mice inactivated for either of the two polymerases related to pol beta (i.e., pol mu and pol lambda) are viable and fertile and display a normal hypermutation pattern. PMID- 11937520 TI - Cutting edge: anti-inflammatory properties of low levels of IFN-gamma. AB - Activation of naive T and B cells occurs only within the context of organized lymphoid tissue. Thus, the continuous recirculation of mature lymphocytes is crucial for the development of primary immune response to foreign Ags. We have previously shown that low levels of IFN-gamma inhibit homing of B cells to the secondary lymphoid organs. In this study, we demonstrate that similarly low doses of IFN-gamma down-regulate integrin-mediated adhesion and migration of naive T and Th2 cells, and have a profound effect on the in vivo homing of naive T cells to the lymph nodes. Moreover, we show that these low doses of IFN-gamma have anti inflammatory effects in an in vivo asthma model. Thus, in contrast to the proinflammatory effects of IFN-gamma at relatively high concentrations, low dose IFN-gamma appears to exert global suppressory effects on T cell trafficking and may have clinical application as an anti-inflammatory agent. PMID- 11937521 TI - Lymphoepithelial interactions trigger specific regulation of gene expression in the M cell-containing follicle-associated epithelium of Peyer's patches. AB - In the intestine, the follicle-associated epithelium (FAE) of Peyer's patches (PP) performs Ag sampling as the first step in developing immune responses. Depending on the species, this epithelium contains 10-50% of M cells, which act as regulated gates in epithelial barriers that can be used opportunistically by pathogens to invade their host. However, the mechanisms involved in the differentiation and uptake processes of M cells are not known, in part because their limited number in the intestinal mucosa has hampered molecular and biochemical studies. In this work we provide evidence that PP lymphocytes can themselves modulate gene expression in PP in vivo and in an in vitro model of FAE. Transgenic mice carrying a reporter gene under the control of a modified L pyruvate kinase promoter (SVPK) exhibit strong transgene expression in PP and FAE, but not in the adjacent villous cells. We used the mouse intestinal epithelial cell line m-IC(cl2) transfected with the SVPK promoter fused to beta galactosidase to investigate the direct effect of PP lymphocytes on SVPK promoter activity. beta-Galactosidase expression was 4.4-fold higher in transfected m IC(cl2) cells when they were cultured with PP lymphocytes. Conversely, green fluorescent protein expression was 1.8-fold lower in stably transfected differentiated intestinal Caco-2(cl1) cells with the sucrase isomaltase promoter fused to green fluorescent protein cDNA when they were cultured with PP lymphocytes, indicating that the in vivo FAE down-regulation of sucrase isomaltase promoter is transcriptionally regulated. PMID- 11937522 TI - I-kappa B kinases alpha and beta have distinct roles in regulating murine T cell function. AB - NF-kappaB is a transcription factor that regulates a variety of genes involved in the control of the immune and inflammatory responses. Activation of NF-kappaB is mediated by an inducible I-kappaB kinase (IKK) complex comprised of two catalytic subunits, IKKalpha and IKKbeta. In this study, the role of these kinases in the development and function of T lymphocytes was explored using transgenic mice expressing the dominant-negative forms of one or both kinases under the control of a T cell-specific promoter. Activation of the NF-kappaB pathway in thymocytes isolated from these transgenic mice following treatment with either PMA and ionomycin or anti-CD3 was markedly inhibited. Although inhibition of IKKalpha and/or IKKbeta function did not alter T cell development in these transgenic mice, the proliferative response to anti-CD3 was reduced in thymocytes isolated from mice expressing dominant-negative IKKbeta. However, inhibition of both IKKalpha and IKKbeta was required to markedly reduce cytokine production in thymocytes isolated from these transgenic mice. Finally, we demonstrated that IKKalpha and IKKbeta have opposite roles on the regulation of anti-CD3-induced apoptosis of double-positive thymocytes. These results suggest that IKKalpha and IKKbeta have distinct roles in regulating thymocyte function. PMID- 11937523 TI - Ultraviolet B radiation induces a transient appearance of IL-4+ neutrophils, which support the development of Th2 responses. AB - UVB irradiation can cause considerable changes in the composition of cells in the skin and in cutaneous cytokine levels. We found that a single exposure of normal human skin to UVB induced an infiltration of numerous IL-4(+) cells. This recruitment was detectable in the papillary dermis already 5 h after irradiation, reaching a peak at 24 h and declining gradually thereafter. The IL-4(+) cells appeared in the epidermis at 24 h postradiation and reached a plateau at days 2 and 3. The number of IL-4(+) cells was markedly decreased in both dermis and epidermis at day 4, and at later time points, the IL-4 expression was absent. The IL-4(+) cells did not coexpress CD3 (T cells), tryptase (mast cells), CD56 (NK cells), and CD36 (macrophages). They did coexpress CD15 and CD11b, showed a clear association with elastase, and had a multilobed nucleus, indicating that UVB induced infiltrating IL-4(+) cells are neutrophils. Blister fluid from irradiated skin, but not from control skin, contained IL-4 protein as well as increased levels of IL-6, IL-8, and TNF-alpha. In contrast to control cultures derived from nonirradiated skin, a predominant type 2 T cell response was detected in T cells present in primary dermal cell cultures derived from UVB-exposed skin. This type 2 shift was abolished when CD15(+) cells (i.e., neutrophils) were depleted from the dermal cell suspension before culturing, suggesting that neutrophils favor type 2 T cell responses in UVB-exposed skin. PMID- 11937524 TI - Polar redistribution of the sialoglycoprotein CD43: implications for T cell function. AB - Contact between T cells and APCs results in the orchestrated segregation of molecules at the cell-cell interface and formation of a specialized structure termed the immunological synapse. This model predicts the topological seclusion of large molecules such as CD43 from the site of closest contact between the T cell and APC, allowing for the close apposition of cell membranes and effective TCR engagement. Similarly, during T cell migration segregation of CD43 to the uropod is thought to aid integrin adhesion at the leading edge of the cell by removing steric hindrance. We show in this work that CD43 distribution on T cells is regulated by a membrane proximal ezrin binding site and that failure to displace CD43 from the immunological synapse has no inhibitory effects on primary T cell activation. We also report that CD43 expression at the contact zone between T cells and matrix does not negatively regulate motility but may regulate LFA-1 de-adhesion. These results suggest that the steric barrier model of CD43 is inadequate and that alternative mechanisms account for the negative regulatory properties of CD43. PMID- 11937525 TI - Selective requirement for CD40-CD154 in drug-induced type 1 versus type 2 responses to trinitrophenyl-ovalbumin. AB - CD154 is transiently expressed by activated T cells and interacts with CD40 on B cells, dendritic cells, macrophages, and monocytes. This costimulatory receptor ligand couple seems decisive in Ag-driven immune responses but may be differentially involved in type 1 vs type 2 responses. We studied the importance of CD40-CD154 in both responses using the reporter Ag popliteal lymph node assay in which selectively acting drugs generate clearly polarized type 1 (streptozotocin) or type 2 (D-penicillamine, diphenylhydantoin) responses to a constant coinjected Ag in the same mouse strain. Treatment of mice with anti CD154 reduced characteristic immunological parameters in type 2 responses (B and CD4(+) T cell proliferation, IgG1 and IgE Abs, and IL-4 secretion) and only slightly affected the type 1 response (small decrease in IFN-gamma production, influx of CD11c(+) and F4/80(+) cells, and prevention of architectural disruption of the lymph node, but no effect on IgG2a Ab and TNF-alpha secretion or B and CD4(+) T cell proliferation). The findings indicate that the CD40-CD154 costimulatory interaction is a prerequisite in drug-induced type 2 responses and is only marginally involved in type 1 responses. The observed expression patterns of CD80 and CD86 on different APC (B cells in type 2 and dendritic cells in type 1) may be responsible for this discrepancy. PMID- 11937526 TI - In vivo triggering through 4-1BB enables Th-independent priming of CTL in the presence of an intact CD28 costimulatory pathway. AB - Triggering of 4-1BB, a member of the TNFR family, through in vivo administration of agonistic anti-4-1BB Ab delivers a powerful costimulatory signal to CTL. We found this signal to effectively replace the need for CD4(+) T cell help in the cross-priming of tumor-specific CTL immunity. Furthermore, 4-1BB Ab can convert an otherwise tolerogenic peptide vaccine into a formulation capable of efficient CTL priming. Initial activation of naive CTL can occur in the absence of 4-1BB costimulation, but this signal permits increased survival of Ag-stimulated CTL. Because naive CTL do not express 4-1BB at their surface, susceptibility to 4-1BB triggering depends on prior up-regulation of this receptor. We show that this requires both stimulation of the TCR and CD28-dependent costimulation. Accordingly, blockade of the CD28-costimulatory pathway abrogates the capacity of agonistic anti-4-1BB Ab to trigger Th-independent CTL immunity. In conclusion, our data reveal that the 4-1BB-mediated survival signal is positioned downstream of Ag-specific TCR triggering and CD28-dependent costimulation of naive CTL. The powerful effects of 4-1BB triggering on the induction, amplification, and persistence of CTL responses provide a novel strategy for increasing the potency of vaccines against cancers. PMID- 11937527 TI - Activating immunity in the liver. II. IFN-beta attenuates NK cell-dependent liver injury triggered by liver NKT cell activation. AB - Dendritic cell (DC)-dependent activation of liver NKT cells triggered by a single i.v. injection of a low dose (10-100 ng/mouse) of alpha-galactosyl ceramide (alphaGalCer) into mice induces liver injury. This response is particularly evident in HBs-tg B6 mice that express a transgene-encoded hepatitis B surface Ag in the liver. Liver injury following alphaGalCer injection is suppressed in mice depleted of NK cells, indicating that NK cells play a role in NK T cell-initiated liver injury. In vitro, liver NKT cells provide a CD80/86-dependent signal to alphaGalCer-pulsed liver DC to release IL-12 p70 that stimulates the IFN-gamma response of NKT and NK cells. Adoptive transfer of NKT cell-activated liver DC into the liver of nontreated, normal (immunocompetent), or immunodeficient (RAG( /-) or HBs-tg/RAG(-/-)) hosts via the portal vein elicited IFN-gamma responses of liver NK cells in situ. IFN-beta down-regulates the pathogenic IL-12/IFN-gamma cytokine cascade triggered by NKT cell/DC/NK cell interactions in the liver. Pretreating liver DC in vitro with IFN-beta suppressed their IL-12 (but not IL 10) release in response to CD40 ligation or specific (alphaGalCer-dependent) interaction with liver NKT cells and down-regulated the IFN-gamma response of the specifically activated liver NKT cells. In vivo, IFN-beta attenuated the NKT cell triggered induction of liver immunopathology. This study identifies interacting subsets of the hepatic innate immune system (and cytokines that up- and down regulate these interactions) activated early in immune-mediated liver pathology. PMID- 11937528 TI - Cells expressing indoleamine 2,3-dioxygenase inhibit T cell responses. AB - Pharmacological inhibition of indoleamine 2,3-dioxygenase (IDO) activity during murine gestation results in fetal allograft rejection and blocks the ability of murine CD8(+) dendritic cells to suppress delayed-type hypersensitivity responses to tumor-associated peptide Ags. These observations suggest that cells expressing IDO inhibit T cell responses in vivo. To directly evaluate the hypothesis that cells expressing IDO inhibit T cell responses, we prepared IDO-transfected cell lines and transgenic mice overexpressing IDO and assessed allogeneic T cell responses in vitro and in vivo. T cells cocultured with IDO-transfected cells did not proliferate but expressed activation markers. The potency of allogeneic T cell responses was reduced significantly when mice were preimmunized with IDO transfected cells. In addition, adoptive transfer of alloreactive donor T cells yielded reduced numbers of donor T cells when injected into IDO-transgenic recipient mice. These outcomes suggest that genetically enhanced IDO activity inhibited T cell proliferation in vitro and in vivo. Genetic manipulation of IDO activity may be of therapeutic utility in suppressing undesirable T cell responses. PMID- 11937529 TI - Temporal segregation of 4-1BB versus CD28-mediated costimulation: 4-1BB ligand influences T cell numbers late in the primary response and regulates the size of the T cell memory response following influenza infection. AB - In this report, we demonstrate that CD28(-/-) mice are severely impaired in the initial expansion of D(b)/NP366-374-specific CD8 T cells in response to influenza virus infection, whereas 4-1BB ligand (4-1BBL)(-/-) mice show no defect in primary T cell expansion to influenza virus. In contrast, 4-1BBL(-/-) mice show a decrease in D(b)/NP366-374-specific T cells late in the primary response. Upon secondary challenge with influenza virus, 4-1BBL(-/-) mice show a decrease in the number of D(b)/NP366-374-specific T cells compared to wild-type mice such that the level of the CD8 T cell expansion during the in vivo secondary response is reduced to the level of a primary response, with concomitant reduction of CTL effector function. In contrast, Ab responses, as well as secondary CD4 T cell responses, to influenza are unaffected by 4-1BBL deficiency. Thus, CD28 is critical for initial T cell expansion, whereas 4-1BB/4-1BBL signaling affects T cell numbers much later in the response and is essential for the survival and/or responsiveness of the memory CD8 T cell pool. PMID- 11937531 TI - Suppression of IL-12 production by soluble CD40 ligand: evidence for involvement of the p44/42 mitogen-activated protein kinase pathway. AB - IL-12 is a key cytokine in skewing immune responses toward Th1-like reactions. Human monocytes/macrophages produce high amounts of bioactive IL-12 when a priming signal (IFN-gamma or GM-CSF) precedes a second signal (e.g., LPS). We and others have previously shown that preincubation with LPS before this stimulation procedure can efficiently and selectively suppress the production of IL-12 by human monocytes. In this study, we show that an almost complete suppression of IL 12 production can also be observed after preincubation of monocytes with costimulatory cell surface molecules that bind to members of the TNFR superfamily (CD40 ligand, TNF-related activation-induced cytokine (TRANCE)). The suppression of IL-12 was observable on the mRNA and protein levels and was not due to endogenous production of known IL-12 antagonists (i.e., IL-10, IL-4, and PGE(2)), to an increased number of cells undergoing apoptosis, nor to down-regulation of the IFN-gamma or CD40 receptor. Cell surface expression of the costimulatory molecules CD80 and CD86 was not reduced by the preincubation procedure, and only a moderate reduction of IL-6 production was observed. Several studies have identified signal transduction pathways that are activated by CD40 signaling, including activation of mitogen-activated protein kinases. The presence of the extracellular signal-related kinase-specific mitogen-activated protein kinase kinase 1/2-specific inhibitors PD98059 and U0126 abrogated suppression induced by sCD40 ligand or other second signals. This indicates that activation of extracellular signal-regulated kinase 1/2 contributes to the underlying mechanism of IL-12 suppression. This mechanism may be relevant in other inflammatory responses and may help to develop therapeutic strategies in Th1-mediated diseases. PMID- 11937530 TI - In vivo CD86 blockade inhibits CD4+ T cell activation, whereas CD80 blockade potentiates CD8+ T cell activation and CTL effector function. AB - To address whether a functional dichotomy exists between CD80 and CD86 in naive T cell activation in vivo, we administered anti-CD80 or CD86 blocking mAb alone or in combination to mice with parent-into-F(1) graft-vs-host disease (GVHD). In this model, the injection of naive parental T cells into unirradiated F(1) mice results in either a Th1 cytokine-driven, cell-mediated immune response (acute GVHD) or a Th2 cytokine-driven, Ab-mediated response (chronic GVHD) in the same F(1) recipient. Combined CD80/CD86 blockade beginning at the time of donor cell transfer mimicked previous results seen with CTLA4Ig and completely abrogated either acute or chronic GVHD by preventing the activation and maturation of donor CD4(+) T cells as measured by a block in acquisition of memory marker phenotype and cytokine production. Similar results were seen with selective CD86 blockade; however, the degree of CD4 inhibition was always less than that seen with combined CD80/CD86 blockade. A more striking effect was seen with selective CD80 blockade in that chronic GVHD was converted to acute GVHD. This effect was associated with the induction of Th1 cytokine production, donor CD8(+) T cell activation, and development of antihost CTL. The similarity of this effect to that reported for selective CTLA4 blockade suggests that CD80 is a critical ligand for CTLA4 in mediating the down-regulation of Th1 responses and CD8(+) T cell activation. In contrast, CD86 is critical for the activation of naive CD4(+) T cells in either a Th1 or a Th2 cytokine-mediated response. PMID- 11937532 TI - A molecular marker for thymocyte-positive selection: selection of CD4 single positive thymocytes with shorter TCRB CDR3 during T cell development. AB - The generation of the naive T cell repertoire is a direct result of maturation and selection events in the thymus. Although maturation events are judged predominantly on the expression of surface markers, molecular markers, more intimately involved in the selection process, can be informative. We have identified a molecular marker for selection in later stages of maturation in humans. Thymocytes are selected for the expression of TCR beta-chains with shorter CDR3 at the double-positive to single-positive (SP) transition. Here we extend these studies to the mouse and show that the selection phenotype is not related to alpha-chain pairing but is a function of the MHC haplotype. Interestingly, the selection is much more apparent in CD4 SP thymocytes than in CD8 SP cells. This is in contrast to human thymocytes, where the selection is equally apparent in both lineages. The involvement of MHC in the process argues that this is a positive selection stage. The difference in the extent of this selection between the two SP lineages may indicate a class difference in the nature of the TCR-MHC interaction, the role of coreceptors in the selection process, or both. PMID- 11937533 TI - Human NK cell-mediated cytotoxicity triggered by CD86 and Gal alpha 1,3-Gal is inhibited in genetically modified porcine cells. AB - Delayed xenograft rejection is a major hurdle that needs to be addressed to prolong graft survival in pig-to-primate xenotransplantation. NK cell activation has been implicated in delayed xenograft rejection. Both Ab-dependent and independent mechanisms are responsible for the high susceptibility of porcine cells to human NK cell-mediated cytotoxicity. Previous reports demonstrated a role of Galalpha1,3-Gal Ag in triggering the Ab-independent responses. We hypothesize that expression of CD80 and/or CD86 on porcine cells may also play a role in NK cell activation as human NK cells express a variant of CD28. Our initial analysis showed that porcine endothelial cells and fibroblasts express CD86, but not CD80. Genetic engineering of these cells to express hCD152-hCD59, a chimeric molecule designed to block CD86 in cis, was accompanied by a reduction in susceptibility to human NK cell-mediated cytotoxicity. The use of a specific anti-porcine CD86-blocking Ab and the NK92 and YTS cell lines further confirmed the involvement of CD86 in triggering NK cell-mediated lysis of porcine cells. Maximal protection was achieved when hCD152-hCD59 was expressed in H transferase transgenic cells, which show reduced Galalpha1,3-Gal expression. In this work, we describe two mechanisms of human NK cell-mediated rejection of porcine cells and demonstrate that genetically modified cells resist Ab-independent NK cell mediated cytotoxicity. PMID- 11937534 TI - Continuous exposure of mice to superantigenic toxins induces a high-level protracted expansion and an immunological memory in the toxin-reactive CD4+ T cells. AB - We analyzed the responses of several T cell fractions reactive with superantigenic toxins (SAGTs), staphylococcal enterotoxin A (SEA), or Yersinia pseudotuberculosis-derived mitogen (YPM) in mice implanted with mini-osmotic pumps filled with SEA or YPM. In mice implanted with the SEA pump, SEA-reactive Vbeta3(+)CD4(+) T cells exhibited a high-level protracted expansion for 30 days, and SEA-reactive Vbeta11(+)CD4(+) T cells exhibited a low-level protracted expansion. SEA-reactive CD8(+) counterparts exhibited only a transient expansion. A similar difference in T cell expansion was also observed in YPM-reactive T cell fractions in mice implanted with the YPM pump. Vbeta3(+)CD4(+) and Vbeta11(+)CD4(+) T cells from mice implanted with the SEA pump exhibited cell divisions upon in vitro restimulation with SEA and expressed surface phenotypes as memory T cells. CD4(+) T cells from mice implanted with the SEA pump exhibited high IL-4 production upon in vitro restimulation with SEA, which was due to the enhanced capacity of the SEA-reactive CD4(+) T cells to produce IL-4. The findings in the present study indicate that, in mice implanted with a specific SAGT, the level of expansion of the SAGT-reactive CD4(+) T cell fractions varies widely depending on the TCR Vbeta elements expressed and that the reactive CD4(+) T cells acquire a capacity to raise a memory response. CD8(+) T cells are low responders to SAGTs. PMID- 11937535 TI - The potency of TCR signaling differentially regulates NFATc/p activity and early IL-4 transcription in naive CD4+ T cells. AB - The potency of TCR signaling can regulate the differentiation of naive CD4(+) T cells into Th1 and Th2 subsets. In this work we demonstrate that TCR signaling by low-affinity, but not high-affinity, peptide ligands selectively induces IL-4 transcription within 48 h of priming naive CD4(+) T cells. This early IL-4 transcription is STAT6 independent and occurs before an increase in GATA-3. Furthermore, the strength of the TCR signal differentially affects the balance of NFATp and NFATc DNA binding activity, thereby regulating IL-4 transcription. Low potency TCR signals result in high levels of nuclear NFATc and low levels of NFATp, which are permissive for IL-4 transcription. These data provide a model for how the strength of TCR signaling can influence the generation of Th1 and Th2 cells. PMID- 11937536 TI - The state of CD4+ T cell activation is a major factor for determining the kinetics and location of T cell responses to oral antigen. AB - Current models suggest that inductive immune responses to enteric Ag are initiated in Peyer's patches (PP) and mesenteric lymph nodes (MLN) followed by migration of activated, memory-like CD4(+) T cells to extralymphoid sites in the intestinal lamina propria (LP). The resultant immune system contains both naive and activated T cells. To examine the differential responses of naive and memory like T cells to oral Ag, bone marrow chimeras (BMC) were generated. Irradiated BALB/c hosts were reconstituted with a mix of DO11.10 x RAG-1(-/-) and BALB/c bone marrow. In unprimed DO11.10 and BMC models, LP and PP DO11.10 T cells responded to oral Ag with similar kinetics. Responses of activated, memory-like T cells to oral Ag were examined in thymectomized BMC 60 days after i.p. immunization with OVA peptide in Freund's adjuvant (OVA(323-339)/CFA). Results indicate that i.p. OVA(323-339)/CFA generated a high proportion of memory-like CD45RB(low) DO11.10 T cells in peripheral lymphoid (40%) and intestinal LP (70%) tissue. Previously activated DO11.10 T cells in the LP responded to oral Ag earlier and at 50% higher levels compared with memory CD4(+) T cells localized to PP tissue. These data indicate that responses to oral Ag in antigenically naive animals are initiated in PP whereas in Ag-experienced animals LP T cells respond earlier and more vigorously than cells in PP. Taken together, these data suggest that previous activation alters the hierarchy of T cell responses to oral Ag by enhancing the efficiency of LP T cell activation. PMID- 11937537 TI - Formation of the killer Ig-like receptor repertoire on CD4+CD28null T cells. AB - Killer Ig-like receptors (KIRs) are expressed on CD4(+)CD28(null) T cells, a highly oligoclonal subset of T cells that is expanded in patients with rheumatoid arthritis. It is unclear at what stage of development these T cells acquire KIR expression. To determine whether KIR expression is a consequence of clonal expansion and replicative senescence, multiple CD4(+)CD28(null) T cell clones expressing the in vivo dominant TCR beta-chain sequences were identified in three patients and analyzed for their KIR gene expression pattern. Based on sharing of TCR sequences, the clones were grouped into five clone families. The repertoire of KIRs was diverse, even within each clone family; however, the gene expression was not random. Three particular receptors, KIR2DS2, KIR2DL2, and KIR3DL2, had significant differences in gene expression frequencies between the clone families. These data suggest that KIRs are successively acquired after TCR rearrangement, with each clone family developing a dominant expression pattern. The patterns did not segregate with the individual from whom the clones were derived, indicating that peripheral selection in the host environment was not a major shaping force. Several models were examined using a computer algorithm that was designed to simulate the expression of KIRs at various times during T cell proliferation. The computer simulations favored a model in which KIR gene expression is inducible for a limited time during the initial stages of clonal expansion. PMID- 11937538 TI - Molecular mechanisms underlying differential contribution of CD28 versus non-CD28 costimulatory molecules to IL-2 promoter activation. AB - T cell costimulation via CD28 and other (non-CD28) costimulatory molecules induces comparable levels of [(3)H]TdR incorporation, but fundamentally differs in the contribution to IL-2 production. In this study, we investigated the molecular basis underlying the difference between CD28 and non-CD28 costimulation for IL-2 gene expression. Resting T cells from a mutant mouse strain generated by replacing the IL-2 gene with a cDNA encoding green fluorescent protein were stimulated with a low dose of anti-CD3 plus anti-CD28 or anti-non-CD28 (CD5 or CD9) mAbs. CD28 and non-CD28 costimulation capable of inducing potent [(3)H]TdR uptake resulted in high and marginal levels of green fluorescent protein expression, respectively, indicating their differential IL-2 promoter activation. CD28 costimulation exhibited a time-dependent increase in the binding of transcription factors to the NF-AT and NF-kappaB binding sites and the CD28 responsive element of the IL-2 promoter, whereas non-CD28 costimulation did not. Particularly, a striking difference was observed for the binding of NF-kappaB to CD28-responsive element and the NF-kappaB binding site. Decreased NF-kappaB activation in non-CD28 costimulation resulted from the failure to translocate a critical NF-kappaB member, c-Rel, to the nuclear compartment due to the lack of IkappaBbeta inactivation. These observations suggest that unlike CD28 costimulation, non-CD28 costimulation fails to sustain IL-2 promoter activation and that such a failure is ascribed largely to the defect in the activation of c Rel/NF-kappaB. PMID- 11937539 TI - Differential roles for extracellularly regulated kinase-mitogen-activated protein kinase in B cell antigen receptor-induced apoptosis and CD40-mediated rescue of WEHI-231 immature B cells. AB - One of the major unresolved questions in B cell biology is how the B cell Ag receptor (BCR) differentially signals to transduce anergy, apoptosis, proliferation, or differentiation during B cell maturation. We now report that extracellularly regulated kinase-mitogen-activated protein kinase (Erk-MAP kinase) can play dual roles in the regulation of the cell fate of the immature B cell lymphoma, WEHI-231, depending on the kinetics and context of Erk-MAP kinase activation. First, we show that the BCR couples to an early (< or =2 h) Erk-MAP kinase signal which activates a phospholipase A(2) pathway that we have previously shown to mediate collapse of mitochondrial membrane potential, resulting in depletion of cellular ATP and cathepsin B execution of apoptosis. Rescue of BCR-driven apoptosis by CD40 signaling desensitizes such early extracellularly regulated kinase (Erk) signaling and hence uncouples the BCR from the apoptotic mitochondrial phospholipase A(2) pathway. A second role for Erk-MAP kinase in promoting the growth and proliferation of WEHI-231 immature B cells is evidenced by data showing that proliferating and CD40-stimulated WEHI-231 B cells exhibit a sustained cycling pattern (8-48 h) of Erk activation that correlates with cell growth and proliferation. This growth-promoting role for Erk signaling is supported by three key pieces of evidence: 1) signaling via the BCR, under conditions that induce growth arrest, completely abrogates sustained Erk activation; 2) CD40-mediated rescue from growth arrest correlates with restoration of cycling Erk activation; and 3) sustained inhibition of Erk prevents CD40-mediated rescue of BCR-driven growth arrest of WEHI-231 immature B cells. Erk-MAP kinase can therefore induce diverse biological responses in WEHI 231 cells depending on the context and kinetics of activation. PMID- 11937540 TI - Negligible class II MHC presentation of B cell receptor-derived peptides by high density resting B cells. AB - Resting B lymphocytes have been credited with inducing T cell tolerance to Ig derived and monovalent self-Ags that are internalized via the B cell receptor (BCR). These conclusions are predicated upon the assumptions that resting B cells display BCR-associated peptides in class II MHC and that the cells remain quiescent during the course of experimental manipulation. To determine whether resting B cells display BCR-associated epitopes in class II MHC, we devised a sensitive assay that averted potential activation of B cells by Ag and minimized activation by prolonged culture. Ex vivo, Percoll-fractionated B cells expressing a kappa transgene encoding a T cell epitope were cultured with a reactive T cell hybridoma for 12 h. Whereas low density, LPS-activated, and BCR-activated B cells elicited significant IL-2 from the T cell hybridoma, resting high density B cells did not. Parallel results were obtained with normal B cells expressing a second epitope encoded by an endogenous V(H) gene. Anergic B cells, which are uniformly low density, also significantly stimulated the T cell hybridoma. Finally, longer culture periods with normal B cells resulted in a higher degree of B cell activation and significant stimulation of reactive T cell hybridomas. Our results provide evidence that activation of B cells profoundly enhances the processing and presentation of BCR-associated Ags. PMID- 11937541 TI - Maturation and activation of dendritic cells induced by lymphocyte activation gene-3 (CD223). AB - Lymphocyte activation gene-3 (LAG-3) is an MHC class II ligand expressed on activated T and NK cells. A LAG-3Ig fusion protein has been used in mice as an adjuvant protein to induce antitumor responses and specific CD8 and CD4 Th1 responses to nominal Ags. In this work we report on the effect of LAG-3Ig on the maturation and activation of human monocyte-derived dendritic cells (DC). LAG-3Ig binds MHC class II molecules expressed in plasma membrane lipid rafts on immature human DC and induces rapid morphological changes, including the formation of dendritic projections. LAG-3Ig markedly up-regulates the expression of costimulatory molecules and the production of IL-12 and TNF-alpha. Consistent with this effect on DC maturation, LAG-3Ig disables DC in their capacity to capture soluble Ags. These events are associated with the acquisition of professional APC function, because LAG-3Ig increases the capacity of DC to stimulate the proliferation and IFN-gamma response by allogeneic T cells. These effects were not observed when using ligation of MHC class II by specific mAb. Class II-mediated signals induced by a natural ligand, LAG-3, lead to complete maturation of DC, which acquire the capacity to trigger naive T cells and drive polarized Th1 responses. PMID- 11937542 TI - The complementarity-determining region-like loops of CD8 alpha interact differently with beta 2-microglobulin of the class I molecules H-2Kb and thymic leukemia antigen, while similarly with their alpha 3 domains. AB - The murine CD8 glycoprotein interacts with both classical MHC class I molecules and some nonclassical molecules, including the thymic leukemia Ag (TL). TL binds preferentially to CD8alphaalpha homodimers with a 10-fold higher affinity than H 2K(b) class I molecules. To understand the molecular basis for this difference, we created a panel of CD8alpha mutants and tested the ability of the CD8alphaalpha homodimers to bind to H-2K(b) tetramers and TL tetramers. Mutations in three CD8 residues located on the complementarity-determining region-like loops contacting the negatively charged loop in the alpha3 domain of MHC class I greatly reduced binding to both tetramers. Because TL and H-2K(b) class I sequences are highly conserved in the alpha3 domain of MHC class I, this suggests that CD8 contacts the alpha3 domain of TL and H-2K(b) in a similar manner. In contrast, mutations in residues on the A and B beta strands of CD8 that are involved in contact with beta(2)-microglobulin affected interaction with the H 2K(b) tetramer, but not the TL tetramer. Therefore, the orientation of interaction of TL with CD8 appears to be different from that of H-2K(b). The unique high affinity binding of TL with CD8alphaalpha is most likely a result of amino acid differences in the alpha3 domain between TL and H-2K(b), particularly at positions 198 (K to D) and 228 (M to T), which are contact residues in the CD8alphaalpha-H-2K(b) cocrystal. PMID- 11937543 TI - During differentiation of the monocytic cell line U937, Pur alpha mediates induction of the CD11c beta 2 integrin gene promoter. AB - CD11c is a member of the beta(2) integrin family of adhesion molecules that, together with CD18, forms a heterodimeric receptor on the surface of myeloid, NK, dendritic, and certain leukemic, lymphoma, and activated lymphoid cells. Monocytic differentiation is associated with an induction of both CD11c and CD18 gene expression. The resulting CD11c/CD18 receptor mediates firm adhesion to the vascular endothelium, transendothelial migration, chemotaxis, and phagocytosis. Monocytic differentiation can be mimicked in vitro by treatment of the promonocytic cell line U937 with PMA. Recently, we reported that in U937 cells, expression of the CD11c gene is controlled by an unidentified transcription factor that binds ssDNA. This finding suggested that DNA secondary structure plays an important role in controlling the CD11c gene and prompted us to search for additional ssDNA-binding activities with which this gene interacts. In this study, we report that in U937 cells, expression of the CD11c gene is mediated by the ssDNA-binding protein Puralpha. During PMA-induced differentiation, the ability of Puralpha to activate the CD11c promoter in U937 cells increases, as does that of Sp1. Together, these increases in the functional activity of both Puralpha and Sp1 combine to induce CD11c expression. PMID- 11937544 TI - Complementarity-determining region 1 sequence requirements drive limited V alpha usage in response to influenza hemagglutinin 307-319 peptide. AB - We have developed a T cell activation-based system that allows for the selection of TCRs with defined peptide/MHC specificities from libraries in which complementarity-determining region (CDR) sequences have been randomized by in vitro mutagenesis. Using this system, we have explored the sequence requirements for CDR1 and CDR2 of the TCR alpha-chain in a human T cell response characterized by restricted Valpha and Vbeta usage. Libraries of T cells expressing receptors built on the framework of a TCR specific for the influenza virus peptide hemagglutinin 307-319 presented by HLA-DR4, but with random sequences inserted at CDR1alpha or CDR2alpha, were selected for response to the same peptide/MHC ligand. A wide variety of CDR2alpha sequences were found to be permissive for recognition. Indeed, >25% of T cell clones chosen at random displayed a significant response. In contrast, a similar challenge of a randomized CDR1alpha library yielded only the parental sequence, and then only after multiple rounds of selection. T cell clones cross-reactive on closely related HLA alleles (subtypes of DR4) could be isolated from randomized libraries, but not clones restricted by more distantly related alleles such as HLA-DR1. These results indicate that, in the context of this T cell response, the structural requirements for recognition at CDR1alpha are significantly more restricted than at CDR2alpha. This system for mutation and selection of TCRs in vitro may be of use in engineering T cells with defined specificities for therapeutic applications. PMID- 11937545 TI - Mitochondria-dependent caspase-9 activation is necessary for antigen receptor mediated effector caspase activation and apoptosis in WEHI 231 lymphoma cells. AB - Engagement of the B cell Ag receptor (BCR) on immature B cells leads to growth arrest followed by apoptosis. Concomitant signaling through CD40 sustains proliferation and rescues the cells from apoptosis. Previously, we have shown that cross-linking CD40 on B cells stimulates the expression of A1, an antiapoptotic member of the Bcl-2 family, and that transduction of the murine B lymphoma line WEHI 231, a model for immature B cells, with A1 protected the cells against BCR-induced apoptosis. Here we demonstrate that A1 strongly interferes with activation of caspase-7, the major effector caspase activated after BCR cross-linking on WEHI 231 lymphoma cells. The pathway leading to activation of the effector caspase cascade including caspase-7 is unclear. Using retrovirally transduced WEHI 231 cell populations, we show that a catalytically inactive mutant of caspase-7 is cleaved almost as efficiently as the wild-type form, arguing against autocatalysis as the sole activating process. In contrast, overexpression of catalytically inactive caspase-9 strongly interferes with caspase-7 processing, poly(ADP-ribose) polymerase cleavage, and DNA laddering, suggesting a role for caspase-9 and hence for the mitochondrial pathway. The importance of the mitochondrial/caspase-9 pathway for BCR-triggered apoptosis is highlighted by our finding that both A1 and the mutant caspase-9 attenuate BCR induced apoptosis. Thus, our data suggest that the BCR-mediated apoptotic signal in immature B cells spreads via a mitochondrial/caspase-9 pathway. PMID- 11937546 TI - Regulation of IL-1 receptor-associated kinases by lipopolysaccharide. AB - IL-1R-associated kinase (IRAK) plays a pivotal role in IL-1R/Toll-like receptor (TLR)-mediated signaling and NF-kappaB activation. IRAK from leukocytes undergoes rapid activation and inactivation/degradation following IL-1 or LPS stimulation. The rapid degradation of IRAK may serve as a negative feedback mechanism of down regulating IL-1R/TLR-mediated signaling and cytokine gene transcription. Although IL-1/IL-1R-triggered IRAK degradation has been studied in detail, the mechanism of LPS-induced IRAK activation and degradation is not clearly defined. In this study, we demonstrate that the IRAK N-terminal 186-aa region is required for LPS induced degradation. The N-terminally truncated IRAK protein expressed in human monocytic THP-1 cells remains stable upon LPS challenge. In comparison, IRAK as well as the IRAK mutant with C-terminal truncation undergo degradation with LPS stimulation. We demonstrate that pretreatment with protein kinase C inhibitor calphostin inhibits LPS-induced IRAK degradation. Furthermore, we observe coimmunoprecipitation of endogenous IRAK and protein kinase C-zeta protein. We show that functional TLR4 is required for LPS-mediated IRAK degradation. IRAK protein in the murine GG2EE cells harboring a mutated TLR4 gene does not undergo degradation upon LPS treatment. In sharp contrast, we observe that the IRAK homolog, IRAK2, does not undergo degradation upon prolonged LPS treatment, suggesting complex regulation of the innate immunity network upon microbial challenge. PMID- 11937548 TI - An analysis of T cell intrinsic roles of E2A by conditional gene disruption in the thymus. AB - The importance of E2A transcription factors in T cell development has been demonstrated in studies of E2A-deficient mice, which display abnormal T cell development and a high frequency of T cell lymphomas. Because E2A expression is not restricted to the T cell lineage, the primary cause of the T cell phenotype in E2A-deficient mice was not fully determined. To further investigate the role of E2A in T cell lineage, we generated mice with the E2A gene disrupted exclusively during thymocyte development using the Cre-lox system. We show that this system allows E2A gene disruption to occur throughout the double-negative stage of thymocyte development. E2A deletion appears to be completed before development reaches the double-positive stage. Consistent with the gene disruption, these mice reveal a T cell intrinsic role for E2A during the transition from the double-negative stage to the double-positive stage of thymocyte development. In contrast to germline E2A knockout mice, conditional E2A knockout mice do not develop T cell lymphoma. This work establishes a new model for further investigating E2A function in T cell development and leukemiogenesis. PMID- 11937547 TI - Identification of a candidate regulatory region in the human CD8 gene complex by colocalization of DNase I hypersensitive sites and matrix attachment regions which bind SATB1 and GATA-3. AB - To locate elements regulating the human CD8 gene complex, we mapped nuclear matrix attachment regions (MARs) and DNase I hypersensitive (HS) sites over a 100 kb region that included the CD8B gene, the intergenic region, and the CD8A gene. MARs facilitate long-range chromatin remodeling required for enhancer activity and have been found closely linked to several lymphoid enhancers. Within the human CD8 gene complex, we identified six DNase HS clusters, four strong MARs, and several weaker MARs. Three of the strong MARs were closely linked to two tissue-specific DNase HS clusters (III and IV) at the 3' end of the CD8B gene. To further establish the importance of this region, we obtained 19 kb of sequence and screened for potential binding sites for the MAR-binding protein, SATB1, and for GATA-3, both of which are critical for T cell development. By gel shift analysis we identified two strong SATB1 binding sites, located 4.5 kb apart, in strong MARs. We also detected strong GATA-3 binding to an oligonucleotide containing two GATA-3 motifs located at an HS site in cluster IV. This clustering of DNase HS sites and MARs capable of binding SATB1 and GATA-3 at the 3' end of the CD8B gene suggests that this region is an epigenetic regulator of CD8 expression. PMID- 11937549 TI - Fine specificity of TCR complementarity-determining region residues and lipid antigen hydrophilic moieties in the recognition of a CD1-lipid complex. AB - alphabeta TCR can recognize peptides presented by MHC molecules or lipids and glycolipids presented by CD1 proteins. Whereas the structural basis for peptide/MHC recognition is now clearly understood, it is not known how the TCR can interact with such disparate molecules as lipids. Recently, we demonstrated that the alphabeta TCR confers specificity for both the lipid Ag and CD1 isoform restriction, indicating that the TCR is likely to recognize a lipid/CD1 complex. We hypothesized that lipids may bind to CD1 via their hydrophobic alkyl and acyl chains, exposing the hydrophilic sugar, phosphate, and other polar functions for interaction with the TCR complementarity-determining regions (CDRs). To test this model, we mutated the residues in the CDR3 region of the DN1 TCR beta-chain that were predicted to project between the CD1b alpha helices in a model of the TCR/CD1 complex. In addition, we tested the requirement for the negatively charged and polar functions of mycolic acid for Ag recognition. Our findings indicate that the CDR loops of the TCR form the Ag recognition domain of CD1 restricted TCRs and suggest that the hydrophilic domains of a lipid Ag can form a combinatorial epitope recognized by the TCR. PMID- 11937550 TI - Generation of cytotoxic T cells against virus-infected human brain macrophages in a murine model of HIV-1 encephalitis. AB - HIV-1 encephalitis (HIVE) and its associated dementia can occur in up to 20% of infected individuals, usually when productive viral replication in brain mononuclear phagocytes (macrophages and microglia) and depletion of CD4(+) T lymphocytes are most significant. T cells control viral replication through much of HIV-1 disease, but how this occurs remains incompletely understood. With this in mind, we studied HIV-1-specific CTL responses in a nonobese diabetic (NOD) SCID mouse model of HIVE. HIV-1-infected monocyte-derived macrophages (MDM) were injected into the basal ganglia after syngeneic immune reconstitution by HLA A*0201-positive human PBL to generate a human PBL-NOD-SCID HIVE mouse. Engrafted T lymphocytes produced HIV-1gag- and HIV-1pol-specific CTL against virus-infected brain MDM within 7 days. This was demonstrated by tetramer staining of human PBL in mouse spleens and by IFN-gamma ELISPOT. CD8, granzyme B, HLA-DR, and CD45R0 Ag reactive T cells and CD79alpha-positive B cells migrated to and were in contact with human MDM in brain areas where infected macrophages were abundant. The numbers of productively infected MDM were markedly reduced (>85%) during 2 wk of observation. The human PBL-NOD-SCID HIVE mouse provides a new tool for studies of cellular immune responses against HIV-1-infected brain mononuclear phagocytes during natural disease and after vaccination. PMID- 11937551 TI - Enhanced Th1 response to Staphylococcus aureus infection in human lactoferrin transgenic mice. AB - Lactoferrin (Lf) is an iron-binding protein of external secretions and neutrophil secondary granules with antimicrobial and immunomodulatory activities. To further define these properties of Lf, we have investigated the response to Staphylococcus aureus infection in transgenic mice carrying a functional human Lf gene. The transgenic mice cleared bacteria significantly better than congenic littermates, associated with a trend to reduced incidence of arthritis, septicemia, and mortality. We identified two pathways by which S. aureus clearance was enhanced. First, human Lf directly inhibited the growth of S. aureus LS-1 in vitro. Second, S. aureus-infected transgenic mice exhibited enhanced Th1 immune polarization. Thus, spleen cells from infected transgenic mice produced higher levels of TNF-alpha and IFN-gamma and less IL-5 and IL-10 upon stimulation ex vivo with the exotoxin toxic shock syndrome toxin-1 compared with congenic controls. To confirm that these effects of Lf transgene expression could occur in the absence of live bacterial infection, we also showed that Lf transgenic DBA/1 mice exhibited enhanced severity of collagen-induced arthritis, an established model of Th1-induced articular inflammation. Higher levels of stainable iron in the spleens of transgenic mice correlated with human Lf distribution, but all other parameters of iron metabolism did not differ between transgenic mice and wild-type littermates. These results demonstrate that human Lf can mediate both antimicrobial and immunomodulatory activities with downstream effects on the outcome of immune pathology in infectious and inflammatory disease. PMID- 11937552 TI - Antibody-mediated control of persistent gamma-herpesvirus infection. AB - The human gamma-herpesviruses, EBV and Kaposi's sarcoma-associated herpesvirus, establish life-long latency and can reactivate in immunocompromised individuals. T cells play an important role in controlling persistent EBV infection, whereas a role for humoral immunity is less clear. The murine gamma-herpesvirus-68 has biological and structural similarities to the human gamma-herpesviruses, and provides an important in vivo experimental model for dissecting mechanisms of immune control. In the current studies, CD28(-/-) mice were used to address the role of Abs in control of persistent murine gamma-herpesvirus-68 infection. Lytic infection was controlled in the lungs of CD28(-/-) mice, and latency was maintained in B cells at normal frequencies. Although class-switched virus specific Abs were initially generated in the absence of germinal centers, titers and viral neutralizing activity rapidly waned. T cell depletion in CD28(-/-) mice with compromised Ab responses, but not in control mice with intact Ab responses, resulted in significant recrudescence from latency, both in the spleen and the lung. Recrudescence could be prevented by passive transfer of immune serum. These data directly demonstrate an important contribution of humoral immunity to control of gamma-herpesvirus latency, and have significant implications for clinical intervention. PMID- 11937553 TI - Trypanosoma cruzi infection selectively renders parasite-specific IgG+ B lymphocytes susceptible to Fas/Fas ligand-mediated fratricide. AB - The control of B cell expansion has been thought to be solely regulated by T lymphocytes. We show in this study that Trypanosoma cruzi infection induces up regulation of both Fas and Fas ligand (FasL) molecules on B cells and renders them susceptible to B cell-B cell killing (referred to as fratricide throughout this paper) mediated via Fas/FasL. Moreover, by in vivo administration of anti FasL blocking mAb we demonstrate that Fas-mediated B cell apoptosis is an ongoing process during this parasitic infection. We also provide evidence that B cells that have switched to IgG isotype are the preferential targets of B cell fratricide. More strikingly, this death pathway selectively affects IgG(+) B cells reactive to parasite but not self Ags. Parasite-specific but not self reactive B cells triggered during this response are rescued after either in vitro or in vivo FasL blockade. Fratricide among parasite-specific IgG(+) B lymphocytes could impair the immune control of T. cruzi and possibly other chronic protozoan parasites. Our results raise the possibility that the blockade of Fas/FasL interaction in the B cell compartment of T. cruzi-infected mice may provide a means for enhancing antiparasitic humoral immune response without affecting host tolerance. PMID- 11937554 TI - Role of NADPH oxidase in the mechanism of lung neutrophil sequestration and microvessel injury induced by Gram-negative sepsis: studies in p47phox-/- and gp91phox-/- mice. AB - We addressed the role of O(2) generated by the NADPH oxidase complex in the mechanism of polymorphonuclear leukocyte (PMN) accumulation and transalveolar migration and lung microvascular injury. Studies were made in mice lacking the p47(phox) and gp91(phox) subunits of NADPH oxidase (p47(phox-/-) and gp91(phox-/ )) in which PMN are incapable of the respiratory burst. The mice were challenged i.p. with live Escherichia coli to induce sepsis. We observed time-dependent increases in PMN sequestration and migration from 1 to 6 h after challenge with 2 x 10(8) E. coli. The responses in knockout mice were greater post-E. coli challenge compared with control mice; i.e., transalveolar PMN migration post-E. coli challenge increased by approximately 50% in the null mice above values in wild type. The increased PMN infiltration was associated with decreased lung bacterial clearance. The generation of the chemoattractant macrophage inflammatory protein-2 in lung tissue was greater in NADPH oxidase-defective mice after E. coli challenge than control mice; moreover, macrophage-inflammatory protein-2 Ab pretreatment prevented the PMN infiltration. We also observed that E. coli failed to increase lung microvascular permeability in p47(phox-/-) and gp91(phox-/-) mice despite the greater lung PMN sequestration. Thus, O(2) production is required for the induction of sepsis-induced lung microvascular injury. We conclude that NADPH oxidase-derived O(2) generation has an important bactericidal role, such that an impairment in bacterial clearance in NADPH oxidase-defective mice results in increased chemokine generation and lung tissue PMN infiltration. PMID- 11937555 TI - Immunization with a recombinant adenovirus encoding a lymphoma idiotype: induction of tumor-protective immunity and identification of an idiotype-specific T cell epitope. AB - The Ig Id of a B cell lymphoma is a tumor-specific Ag, although as a self-Ag it is likely to be a weak immunogen. Provision of a foreign gene may enhance the immunogenicity of the idiotype. Viral vectors allow highly efficient transfer of genetic material and are themselves innately immunogenic. We have investigated the ability of recombinant adenoviral vectors, encoding the idiotypic gene with or without fusion to the human Fc region, to produce anti-idiotypic Ab- and T cell-mediated responses in a syngeneic BALB/c A20 murine lymphoma model. The idiotypic V(H) and V(L) sequences were assembled as a single chain variable fragment (scFv) and adenoviral vectors encoding the A20 scFv (Ad.A20) and A20 scFv linked to the Fc fragment of human IgG1 (Ad.A20hFc) were constructed. A single immunization of BALB/c mice with Ad.A20hFc but not Ad.A20 induced a specific anti-idiotypic Ab response. T cell lines generated from mice vaccinated with either vector displayed specific cytotoxicity, proliferation, and IFN-gamma release against a syngeneic dendritic cell line transduced using a retroviral vector to express the A20 scFv idiotype (XS52.A1.A20). Importantly, both T cell lines lysed the A20 lymphoma cells. An immunodominant H-2K(d)-restricted CD8(+) T cell peptide, DYWGQGTEL (A20[106-114]), was identified as a naturally occurring A20 scFv epitope. A single immunization with Ad.A20hFc but not Ad.A20 provided protection in >40% of animals challenged with a lethal dose of the A20 tumor line and was more effective, in this model, than a previously optimized plasmid vaccine. PMID- 11937556 TI - CD8+ T cells are required for primary immunity in C57BL/6 mice following low dose, intradermal challenge with Leishmania major. AB - Standard murine models of cutaneous leishmaniasis, involving s.c. inoculation of large numbers of Leishmania major promastigotes, have not supported an essential role for CD8(+) T cells in the control of primary infection. Recently, a L. major model combining two main features of natural transmission, low parasite dose and inoculation into a dermal site, has been established in resistant C57BL/6 mice. In the present studies, C57BL/6 mice with CD8(+) T cell deficiencies, including CD8(-/-) and CD8-depleted mice, failed to control the growth of L. major following inoculation of 100 metacyclic promastigotes into the ear dermis. The resulting dermal pathology was minor and delayed. Lesion formation in wild-type mice was coincident with the killing of parasites in the inoculation site. Both events were associated with the accumulation of CD8(+) T lymphocytes in the skin and with the capacity of CD8(+) T cells recovered from draining lymph nodes or infected dermis to release IFN-gamma following coculture with infected dendritic cells. Reconstitution of resistance to L. major in RAG(-/-) mice using T cells from naive donors was optimal when both CD4(+) and CD8(+) T cells were transferred. Primed CD8(+) T lymphocytes obtained from C57BL/6 mice during the acute stage of infection were able to mediate both pathology and immunity when transferred alone. The low dose, intradermal challenge model reveals that CD8(+) T cells play an essential role in both pathogenesis of and immunity to primary infection with L. major in the skin. PMID- 11937557 TI - Detailed analysis of CD4+ Th responses to envelope and Gag proteins of simian immunodeficiency virus reveals an exclusion of broadly reactive Th epitopes from the glycosylated regions of envelope. AB - Ag-specific CD4(+) Th cells play a key role in the development, maturation, and maintenance of pathogen-specific humoral and cellular immune responses. To define the fine specificity of broadly reactive Th responses associated with mature immunity in a lentiviral system, we analyzed peptide-specific Th responses in eight macaques chronically infected with a reference live attenuated SIV at 12-14 mo postinoculation. All macaques had stable immunocompetent Th cells at the time of analysis, and a unique array of Th responses to 20-mer overlapping peptides from envelope (Env) and Gag was identified for each macaque, which were then used to define a set of 31 broadly reactive peptide epitopes. Only 5 of the 31 broadly reactive Th epitope peptides mapped to the surface (SU) domain of Env. Interestingly, these were all confined to two conserved nonglycosylated regions toward the carboxyl terminus of SU, suggesting a structural influence of glycosylation on development of Th responses. Gag and the Env transmembrane proteins contained the majority of broadly reactive peptide epitopes (12 and 14 peptides, respectively), which were uniformly distributed throughout their sequence. This study defines for the first time broadly reactive Th epitope peptides of SIV Env and Gag proteins that are associated with enduring broadly protective vaccine immunity to attenuated SIV, which may be used for the design and evaluation of experimental vaccines. Moreover, the data suggest that extensive glycosylation of SU may provide yet another immune escape mechanism developed by lentiviruses to restrict the breadth of Th repertoire to SU, a major immunologically exposed protein of the virus. PMID- 11937558 TI - Two lipoproteins extracted from Escherichia coli K-12 LCD25 lipopolysaccharide are the major components responsible for Toll-like receptor 2-mediated signaling. AB - Toll-like receptor 2 (TLR2)-mediated cell activation induced by commercial preparations of LPS was recently shown to arise from impurities whose identities are not known. In this work, we determined the molecules responsible for TLR2 mediated cell activation in LPS derived from Escherichia coli K-12 strain LCD25. When LCD25 LPS was phenol extracted, two proteins capable of TLR2-mediated cell activation were purified and identified as E. coli lipoproteins. We cloned, expressed, and purified these two lipoproteins, Lip19 and Lip12. Lip19 or Lip12 activated TNF-alpha production from RAW264.7 and THP-1 cells in a TLR2-dependent manner. However, neither Lip19 nor Lip12 activated HUVECs, which lack endogenous TLR2. Additionally, IkappaB kinase beta and c-Jun N-terminal kinase 1 activation in THP-1 cells induced by Lip19 or Lip12 was observed. TLR2 activation by Lip19 and Lip12 in HEK293 cells was blocked by inhibitory anti-TLR2 mAbs. The unlipidated mutants, Lip19-C19S and Lip12-C21S, in which the NH(2)-terminal cysteine was substituted by serine, lost their ability to activate TLR2 transfected HEK 293 cells. Taken together, these results demonstrate that two lipoproteins constitute the major contaminants responsible for TLR2-mediated cell activation in E. coli LCD25 LPS. PMID- 11937559 TI - Leptin-deficient mice exhibit impaired host defense in Gram-negative pneumonia. AB - Leptin is an adipocyte-derived hormone that is secreted in correlation with total body lipid stores. Serum leptin levels are lowered by the loss of body fat mass that would accompany starvation and malnutrition. Recently, leptin has been shown to modulate innate immune responses such as macrophage phagocytosis and cytokine synthesis in vitro. To determine whether leptin plays a role in the innate host response against Gram-negative pneumonia in vivo, we compared the responses of leptin-deficient and wild-type mice following an intratracheal challenge of Klebsiella pneumoniae. Following K. pneumoniae administration, we observed increased leptin levels in serum, bronchoalveolar lavage fluid, and whole lung homogenates. In a survival study, leptin-deficient mice, as compared with wild type mice, exhibited increased mortality following K. pneumoniae administration. The increased susceptibility to K. pneumoniae in the leptin-deficient mice was associated with reduced bacterial clearance and defective alveolar macrophage phagocytosis in vitro. The exogenous addition of very high levels of leptin (500 ng/ml) restored the defect in alveolar macrophage phagocytosis of K. pneumoniae in vitro. While there were no differences between wild-type and leptin-deficient mice in lung homogenate cytokines TNF-alpha, IL-12, or macrophage-inflammatory protein-2 after K. pneumoniae administration, leukotriene synthesis in lung macrophages from leptin-deficient mice was reduced. Leukotriene production was restored by the addition of exogenous leptin (500 ng/ml) to macrophages in vitro. This study demonstrates for the first time that leptin-deficient mice display impaired host defense in bacterial pneumonia that may be due to a defect in alveolar macrophage phagocytosis and leukotriene synthesis. PMID- 11937560 TI - Role of p38 mitogen-activated protein kinase phosphorylation and Fas-Fas ligand interaction in morphine-induced macrophage apoptosis. AB - In this study, we evaluated the molecular mechanisms involved in morphine-induced macrophage apoptosis. Both morphine and TGF-beta promoted P38 mitogen-activated protein kinase (MAPK) phosphorylation, and this phosphorylation was inhibited by SB 202190 as well as by SB 203580. Anti-TGF-beta Ab as well as naltrexone (an opiate receptor antagonist) inhibited morphine-induced macrophage P38 MAPK phosphorylation. Anti-TGF-beta Ab also attenuated morphine-induced p53 as well as inducible NO synthase expression; in contrast, N(G)-nitro-L-arginine methyl ester, an inhibitor of NO synthase, inhibited morphine-induced P38 MAPK phosphorylation and Bax expression. Morphine also enhanced the expression of both Fas and Fas ligand (FasL), whereas anti-FasL Ab prevented morphine-induced macrophage apoptosis. Moreover, naltrexone inhibited morphine-induced FasL expression. In addition, macrophages either deficient in FasL or lacking p53 showed resistance to the effect of morphine. Inhibitors of both caspase-8 and caspase-9 partially prevented the apoptotic effect of morphine on macrophages. In addition, caspase-3 inhibitor prevented morphine-induced macrophage apoptosis. These findings suggest that morphine-induced macrophage apoptosis proceeds through opiate receptors via P38 MAPK phosphorylation. Both TGF-beta and inducible NO synthase play an important role in morphine-induced downstream signaling, which seems to activate proteins involved in both extrinsic (Fas and FasL) and intrinsic (p53 and Bax) cell death pathways. PMID- 11937561 TI - Activation of the neutrophil nicotinamide adenine dinucleotide phosphate oxidase by galectin-1. AB - Galectins are a group of lactose-binding proteins widely distributed in nature. Twelve mammalian galectins have so far been identified, but their functions are to a large extent unknown. In this work we study galectin-1 in its interaction with human neutrophils, with regard to both cell surface binding and activation of the superoxide-producing NADPH-oxidase. We show that galectin-1 is able to activate the neutrophil NADPH-oxidase, provided that the cells have been primed by extravasation from the blood into the tissue, an activation pattern that is similar to that of galectin-3. Using in vitro priming protocols, the galectin-1 responsiveness was found to correlate to granule mobilization and galectin-1 binding to the cells, suggesting the presence of granule-localized receptors that are up-regulated to the cell surface upon priming. By galectin-1 overlay of fractionated neutrophils we identified potential galectin-1 receptor candidates localized in the membranes of the secretory vesicle and gelatinase granules. The binding of galectin-1 and galectin-3 to neutrophil proteins was compared, as were the dose dependencies for activation by the two lectins. The results suggest that, although similarities are found between the two galectins, they appear to activate the NADPH-oxidase using different receptors. In conclusion, galectin-1 appears to have proinflammatory functions, mediated through activation of the neutrophil respiratory burst. PMID- 11937562 TI - Early events in the activation of Fc gamma RIIA in human neutrophils: stimulated insolubilization, translocation to detergent-resistant domains, and degradation of Fc gamma RIIA. AB - The signal transduction mechanisms associated with the ligation of FcgammaRIIA in human neutrophils are as yet only incompletely characterized. In the present study, we have investigated the distribution and fate of FcgammaRIIA following its cross-linking. The results obtained indicate that cross-linking of FcgammaRIIA led, within a few seconds, to its translocation into a nonionic detergent-insoluble fraction. This was followed, within a couple of minutes, by a substantial loss of immunoreactive FcgammaRIIA in the cells. The stimulated degradation of FcgammaRIIA was blocked by the Src kinase inhibitor PP1 but not by wortmannin, ST-638, piceatannol, or cytochalasin B. Cross-linked FcgammaRIIA could be solubilized by saponin (in the presence of Nonidet P-40) and by beta octylglucoside. Sucrose gradient analysis of the distribution of FcgammaRIIA revealed that its cross-linking led to its translocation into the pellets and not the light buoyant density fractions classically associated with lipid rafts. Disruption of cholesterol-containing membrane microdomains with filipin prevented the degradation of FcgammaRIIA but did not inhibit the stimulation of the pattern of tyrosine phosphorylation or the mobilization of calcium that followed FcgammaRIIA cross-linking. These data suggest that both cholesterol-rich domains and Src kinases are required for the degradation of the activated FcgammaRIIA and provide new insights into the early events following FcgammaRIIA cross-linking. PMID- 11937563 TI - Early phase bronchoconstriction in the mouse requires allergen-specific IgG. AB - Allergen provocation of allergic asthma patients is often characterized by an initial period of bronchoconstriction, or early phase reaction (EPR), that leads to maximal airway narrowing within 15-30 min, followed by a recovery period returning airway function to baseline within 1-2 h. In this study, we used a defined OVA provocation model and mice deficient for specific leukocyte populations to investigate the cellular/molecular origins of the EPR. OVA sensitized/challenged wild-type (C57BL/6J) mice displayed an EPR following OVA provocation. However, this response was absent in gene knockout animals deficient of either B or T cells. Moreover, transfer of OVA-specific IgG, but not IgE, before the OVA provocation, was capable of inducing the EPR in both strains of lymphocyte-deficient mice. Interestingly, an EPR was also observed in sensitized/challenged mast cell-deficient mice following an OVA provocation. These data show that the EPR in the mouse is an immunologically based pathophysiological response that requires allergen-specific IgG but occurs independent of mast cell activities. Thus, in the mouse the initial period of bronchoconstriction following allergen exposure may involve neither mast cells nor IgE-mediated events. PMID- 11937564 TI - C/EBP beta gene inactivation causes both impaired and enhanced gene expression and inverse regulation of IL-12 p40 and p35 mRNAs in macrophages. AB - The transcription factor C/EBPbeta is believed to play a fundamental role in regulating activated macrophage functions. However, the molecular mechanisms and the target genes involved have been, so far, poorly characterized, partly due to the difficulty of reproducibly obtaining homogeneous and abundant primary macrophage populations. In this study, we describe the generation and characterization of immortalized macrophage-like cell lines from C/EBPbeta deficient and wild-type mice. Using these cells, we were able to identify a number of genes involved in activated macrophage functions whose induction was affected in the C/EBPbeta(-/-) cells. IFN-gamma/LPS-dependent induction of IL-6, IL-1beta, TNF-alpha, inducible NO synthase, and plasminogen activator inhibitor-1 mRNAs was variably impaired, while IL-12 p40, RANTES and macrophage inflammatory protein-1beta mRNAs were up-regulated in the absence of C/EBPbeta. The differential mRNA expression correlated with differential transcription levels of the corresponding genes, and was in most cases confirmed in primary macrophage populations. Moreover, in sharp contrast to the enhanced induction of IL-12 p40 mRNA, C/EBPbeta(-/-) primary macrophages derived from both the bone marrow and the peritoneal cavity displayed totally defective expression of IL-12 p35 mRNA. Therefore, the IL-12 p35 gene represents a novel obligatory target for C/EBPbeta in macrophages and this may explain the defective production of bioactive IL-12 and the impaired Th1 responses of C/EBPbeta-deficient mice to Candida albicans infection observed in previous work. PMID- 11937565 TI - Store-operated calcium entry in human neutrophils reflects multiple contributions from independently regulated pathways. AB - Human polymorphonuclear neutrophil (PMN) responses to G protein-coupled chemoattractants are highly dependent upon store-operated Ca(2+) entry (SOCE). Recent research suggests that SOCE currents can be mediated by a variety of related channel proteins of the transient receptor potential superfamily. SOCE has been regarded as a specific response to depletion of cell calcium stores. We hypothesized that net SOCE might reflect the contributions of more than one calcium entry pathway. SOCE was studied in normal human PMN using Ca(2+) and Sr(2+) ions. We found that PMN SOCE depends on at least two divalent cation influx pathways. One of these was nonspecific and Sr(2+) permeable; the other was Ca(2+) specific. The two pathways show different degrees of dependence on store depletion by thapsigargin and ionomycin, and differential sensitivity to inhibition by 2-aminoethyoxydiphenyl borane and gadolinium. The inflammatory G protein-coupled chemoattractants fMLP, platelet-activating factor, and IL-8 elicit unique patterns of Sr(2+) and Ca(2+) influx channel activation, and SOCE responses to these agonists displayed differing degrees of linkage to prior Ca(2+) store depletion. The mechanisms of PMN SOCE responses to G protein-coupled chemoattractants are physiologically diverse. They appear to reflect Ca(2+) transport through a variety of channels that are independently regulated to varying degrees by store depletion and by G protein-coupled receptor activation. PMID- 11937566 TI - Anti-inflammatory effects of a p38 mitogen-activated protein kinase inhibitor during human endotoxemia. AB - The p38 mitogen-activated protein kinase (MAPK) participates in intracellular signaling cascades resulting in inflammatory responses. Therefore, inhibition of the p38 MAPK pathway may form the basis of a new strategy for treatment of inflammatory diseases. However, p38 MAPK activation during systemic inflammation in humans has not yet been shown, and its functional significance in vivo remains unclear. Hence, we exposed 24 healthy male subjects to an i.v. dose of LPS (4 ng/kg), preceded 3 h earlier by orally administered 600 or 50 mg BIRB 796 BS (an in vitro p38 MAPK inhibitor) or placebo. Both doses of BIRB 796 BS significantly inhibited LPS-induced p38 MAPK activation in the leukocyte fraction of the volunteers. Cytokine production (TNF-alpha, IL-6, IL-10, and IL-1R antagonist) was strongly inhibited by both low and high dose p38 MAPK inhibitor. In addition, p38 MAPK inhibition diminished leukocyte responses, including neutrophilia, release of elastase-alpha(1)-antitrypsin complexes, and up-regulation of CD11b with down-regulation of L-selectin. Finally, blocking p38 MAPK decreased C reactive protein release. These data identify p38 MAPK as a principal mediator of the inflammatory response to LPS in humans. Furthermore, the anti-inflammatory potential of an oral p38 MAPK inhibitor in humans in vivo suggests that p38 MAPK inhibitors may provide a new therapeutic option in the treatment of inflammatory diseases. PMID- 11937567 TI - Regulation of the mannan-binding lectin pathway of complement on Neisseria gonorrhoeae by C1-inhibitor and alpha 2-macroglobulin. AB - We examined complement activation by Neisseria gonorrhoeae via the mannan-binding lectin (MBL) pathway in normal human serum. Maximal binding of MBL complexed with MBL-associated serine proteases (MASPs) to N. gonorrhoeae was achieved at a concentration of 0.3 microg/ml. Preopsonization with MBL-MASP at concentrations as low as 0.03 microg/ml resulted in approximately 60% killing of otherwise fully serum-resistant gonococci. However, MBL-depleted serum (MBLdS) reconstituted with MBL-MASP before incubation with organisms (postopsonization) failed to kill at a 100-fold higher concentration. Preopsonized organisms showed a 1.5-fold increase in C4, a 2.5-fold increase in C3b, and an approximately 25-fold increase in factor Bb binding; enhanced C3b and factor Bb binding was classical pathway dependent. Preopsonization of bacteria with a mixture of pure C1-inhibitor and/or alpha(2)-macroglobulin added together with MBL-MASP, all at physiologic concentrations before adding MBLdS, totally reversed killing in 10% reconstituted serum. Reconstitution of MBLdS with supraphysiologic (24 microg/ml) concentrations of MBL-MASP partially overcame the effects of inhibitors (57% killing in 10% reconstituted serum). We also examined the effect of sialylation of gonococcal lipooligosaccharide (LOS) on MBL function. Partial sialylation of LOS did not decrease MBL or C4 binding but did decrease C3b binding by 50% and resulted in 80% survival in 10% serum (lacking bacteria-specific Abs) even when sialylated organisms were preopsonized with MBL. Full sialylation of LOS abolished MBL, C4, and C3b binding, resulting in 100% survival. Our studies indicate that MBL does not participate in complement activation on N. gonorrhoeae in the presence of "complete" serum that contains C1-inhibitor and alpha(2) macroglobulin. PMID- 11937568 TI - Inhibition of Rho GTPases with protein prenyltransferase inhibitors prevents leukocyte recruitment to the central nervous system and attenuates clinical signs of disease in an animal model of multiple sclerosis. AB - The ICAM-1-mediated brain endothelial cell (EC)-signaling pathway induced by adherent lymphocytes is a central element in facilitating lymphocyte migration through the tight endothelial barrier of the brain. Rho proteins, which must undergo posttranslational prenylation to be functionally active, have been shown to be an essential component of this signaling cascade. In this study, we have evaluated the effect of inhibiting protein prenylation in brain ECs on their ability to support T lymphocyte migration. ECs treated in vitro with protein prenylation inhibitors resulted in a significant reduction in transendothelial T lymphocyte migration. To determine the therapeutic potential of this approach, an animal model of multiple sclerosis, experimental autoimmune encephalomyelitis, was induced in Biozzi ABH mice. Animals treated before disease onset with protein prenylation inhibitors exhibited a dramatic and significant reduction in both leukocyte infiltration into the CNS and clinical presentation of disease compared with untreated animals. These studies demonstrate, for the first time, the potential for pharmacologically targeting CNS EC signaling responses, and particularly endothelial Rho proteins, as a means of attenuating leukocyte recruitment to the CNS. PMID- 11937569 TI - Restricted localization of the TNF receptor CD120a to lipid rafts: a novel role for the death domain. AB - The TNF-alpha receptor, CD120a, has recently been shown to be localized to both plasma membrane lipid rafts and to the trans Golgi complex. Through a combination of both confocal microscopy and sucrose density gradient ultracentrifugation, we show that amino acid sequences located within the death domain (DD) of CD120a are both necessary and sufficient to promote the appropriate localization of the receptor to lipid rafts. Deletion of the DD (CD120a.Delta321-425) prevented the receptor from being targeted to lipid rafts and resulted in a uniform plasma membrane localization. A similar loss of raft localization was also observed following pairwise deletion of the six alpha-helices that comprise the DD. In all situations, the loss of the ability of CD120a to become localized to lipid rafts following mutagenesis was paralleled by a failure of the receptor to initiate apoptosis. Furthermore, introduction of the lpr mutation into CD120a (CD120a.L351N) also resulted in both a loss in the ability of the receptor to signal apoptosis and to be appropriately localized to rafts. In contrast to CD120a, CD120b, which lacks a DD, is mainly expressed in the bulk plasma membrane and to a lesser extent in lipid rafts, but is absent from the Golgi complex. However, a chimeric receptor in which the DD of CD120a was fused to the cytoplasmic domain of CD120b was predominantly localized to lipid rafts. Collectively, these findings suggest that in addition to its role in CD120a signaling, an appropriately folded and functionally active DD is required for the localization of the receptor to lipid rafts. PMID- 11937570 TI - IL-9 inhibits oxidative burst and TNF-alpha release in lipopolysaccharide stimulated human monocytes through TGF-beta. AB - IL-9 is a Th2 cytokine that exerts pleiotropic activities on T cells, B cells, mast cells, hematopoietic progenitors, and lung epithelial cells, but no effect of this cytokine has been reported so far on mononuclear phagocytes. Human blood monocytes preincubated with IL-9 for 24 h before LPS or PMA stimulation exhibited a decreased oxidative burst, even in the presence of IFN-gamma. The inhibitory effect of IL-9 was specifically abolished by anti-hIL-9R mAb, and the presence of IL-9 receptors was demonstrated on human blood monocytes by FACS. IL-9 also down regulated TNF-alpha and IL-10 release by LPS-stimulated monocytes. In addition, IL-9 strongly up-regulated the production of TGF-beta1 by LPS-stimulated monocytes. The suppressive effect of IL-9 on the respiratory burst and TNF-alpha production in LPS-stimulated monocytes was significantly inhibited by anti-TGF beta1, but not by anti-IL-10Rbeta mAb. Furthermore, IL-9 inhibited LPS-induced activation of extracellular signal-regulated kinase 1/2 mitogen-activated protein kinases in monocytes through a TGF-beta-mediated induction of protein phosphatase activity. In contrast, IL-4, which exerts a similar inhibitory effect on the oxidative burst and TNF-alpha release by monocytes, acts primarily through a down regulation of LPS receptors. Thus, IL-9 deactivates LPS-stimulated blood mononuclear phagocytes, and the mechanism of inhibition involves the potentiation of TGF-beta1 production and extracellular signal-regulated kinase inhibition. These findings highlight a new target cell for IL-9 and may account for the beneficial activity of IL-9 in animal models of exaggerated inflammatory response. PMID- 11937571 TI - Monocyte chemoattractant protein-1 and 5-lipoxygenase products recruit leukocytes in response to platelet-activating factor-like lipids in oxidized low-density lipoprotein. AB - Oxidized low-density lipoprotein (LDL) contains inflammatory agents, including oxidatively fragmented phospholipids that activate the platelet-activating factor (PAF) receptor, but in vivo events caused by these pathologically generated agents are not well defined. Injection of PAF-like lipids derived from oxidized LDL, or C(4)-PAF that is a major PAF-like lipid in these particles, into the pleural cavity of mice resulted in rapid monocyte, neutrophil, and eosinophil accumulation. Increased numbers of intracellular lipid bodies in these cells show they were in an inflammatory environment. Leukocyte recruitment was abolished by a PAF receptor antagonist, as expected. PAF-like lipids induced 5-lipoxygenase expression in leukocytes, mRNA expression for monocyte chemoattractant protein-1 (MCP-1) and other chemokines, synthesis of MCP-1, and leukotriene B(4). The 5 lipoxygenase inhibitor zileuton impaired neutrophil influx, while MCP-1 had a more global role, as determined with MCP-1(-/-) mice. The lack of MCP-1 abrogated leukocyte accumulation and lipid body formation both in vivo and in vitro and chemokine transcription in vivo, and reduced in vivo leukotriene B(4) production. Thus, PAF-like phospholipids in oxidized LDL induce an inflammatory infiltrate through the PAF receptor, chemokine transcription, lipid body formation, and 5 lipoxygenase expression in leukocytes. MCP-1 has a key role in this inflammatory response, and 5-lipoxygenase products are essential for neutrophil recruitment into the inflamed pleural cavity. PMID- 11937572 TI - CXCR4 function requires membrane cholesterol: implications for HIV infection. AB - HIV requires cholesterol and lipid rafts on target cell membranes for infection. To elucidate a possible mechanism, we determined that cholesterol extraction by hydroxypropyl-beta-cyclodextrin (BCD) inhibits stromal cell-derived factor 1alpha (SDF-1alpha) binding to CXCR4 on T cell lines and PBMCs. Intracellular calcium responses to SDF-1alpha, as well as receptor internalization, were impaired in treated T cells. Loss in ligand binding is likely due to conformational changes in CXCR4 and not increased sensitivity to internalization. SDF-1alpha binding and calcium responses were effectively restored by reloading cholesterol. Immunofluorescence microscopy revealed that SDF-1alpha binding occurred in lipid raft microdomains that contained GM1. CXCR4 surface expression, on the other hand, only partially colocalized with GM1. HIV-1(IIIB) infection assays confirmed the functional loss of CXCR4 in the cell lines tested, Sup-T1 and CEM-NKR-CCR5. These data suggest that cholesterol is essential for CXCR4 conformation and function and that lipid rafts may play a regulatory role in SDF-1alpha signaling. PMID- 11937573 TI - IL-16 regulation of human mast cells/basophils and their susceptibility to HIV-1. AB - AIDS patients often contain HIV-1-infected mast cells (MCs)/basophils in their peripheral blood, and in vivo-differentiated MCs/basophils have been isolated from the blood of asthma patients that are HIV-1 susceptible ex vivo due to their surface expression of CD4 and varied chemokine receptors. Because IL-16 is a ligand for CD4 and/or an undefined CD4-associated protein, the ability of this multifunctional cytokine to regulate the development of human MCs/basophils from nongranulated progenitors residing in cord or peripheral blood was evaluated. After 3 wk of culture in the presence of c-kit ligand, IL-16 induced the progenitors residing in the blood of normal individuals to increase their expression of chymase and tryptase about 20-fold. As assessed immunohistochemically, >80% of these tryptase(+) and/or chymase(+) cells expressed CD4. The resulting cells responded to IL-16 in an in vitro chemotaxis assay, and this biologic response could be blocked by anti-IL-16 and anti-CD4 Abs as well as by a competitive peptide inhibitor corresponding to a sequence in the C-terminal domain of IL-16. The additional finding that IL-16 induces calcium mobilization in the HMC-1 cell line indicates that IL-16 acts directly on MCs and their committed progenitors. IL-16-treated MCs/basophils also are less susceptible to infection by an M/R5-tropic strain of HIV-1. Thus, IL-16 regulates MCs/basophils at a number of levels, including their vulnerability to retroviral infection. PMID- 11937574 TI - IFN-gamma production by intrinsic renal cells and bone marrow-derived cells is required for full expression of crescentic glomerulonephritis in mice. AB - The contribution of IFN-gamma from bone marrow (BM) and non-BM-derived cells to glomerular and cutaneous delayed-type hypersensitivity (DTH) was studied in mice. Chimeric IFN-gamma mice (IFN-gamma(+/+) BM chimera), in which IFN-gamma production was restricted to BM-derived cells, were created by transplanting normal C57BL/6 (wild-type (WT)) BM into irradiated IFN-gamma-deficient mice. BM IFN-gamma-deficient chimeric mice (IFN-gamma(-/-) BM chimera) were created by transplanting WT mice with IFN-gamma-deficient BM. WT and sham chimeric mice (WT mice transplanted with WT BM) developed crescentic glomerulonephritis (GN) with features of DTH (including glomerular T cell and macrophage infiltration) in response to an Ag planted in their glomeruli and skin DTH following subdermal Ag challenge. IFN-gamma-deficient mice showed significant protection from crescentic GN and reduced cutaneous DTH. IFN-gamma(+/+) BM chimeric and IFN-gamma(-/-) BM chimeric mice showed similar attenuation of crescentic GN as IFN-gamma-deficient mice, whereas cutaneous DTH was reduced only in IFN-gamma(-/-) BM chimeras. In crescentic GN, IFN-gamma was expressed by tubular cells and occasional glomerular cells and was colocalized with infiltrating CD8(+) T cells, but not with CD4(+) T cells or macrophages. Renal MHC class II expression was reduced in IFN-gamma(+/+) BM chimeric mice and was more severely reduced in IFN-gamma-deficient mice and IFN-gamma(-/-) BM chimeric mice. These studies show that IFN-gamma expression by both BM-derived cells and intrinsic renal cells is required for the development of crescentic GN, but IFN-gamma production by resident cells is not essential for the development of cutaneous DTH. PMID- 11937575 TI - Chronic graft-versus-host in Ig knockin transgenic mice abrogates B cell tolerance in anti-double-stranded DNA B cells. AB - Anti-dsDNA Abs are specific diagnostic markers of systemic lupus erythematosus, and are also implicated in kidney pathology. Anti-dsDNA B cells have been shown to be tolerized in nonautoimmune mice. The immunodysregulation that causes these cells to break tolerance is presumably part of the fundamental defects in systemic lupus erythematosus. To explore these mechanisms, we used the chronic graft-versus-host model mediated by MHC class II differences. Induction of chronic graft-vs-host in anti-DNA H chain knockin (3H9.KI) transgenic mice on a nonautoimmune background resulted in specific activation of anti-dsDNA B cells, as evidenced by high titers of soluble Ab in sera and a high frequency (70%) of anti-dsDNA B cell clones recovered as hybridomas. In addition, the lambda(+)-anti dsDNA B cells developed increased expression of cell surface activation markers, and concentrated in the T cell area of the follicle with an Ab-forming cell compatible phenotype. Genetic analysis of the hybridoma clones showed strong evidence of secondary rearrangements of the L chain associated with anti-dsDNA reactivity. Thus, our study indicates that alloreactive T cell help can break tolerance in a complex manner, involving several events. PMID- 11937576 TI - Gene expression profiling in DQA1*0501+ children with untreated dermatomyositis: a novel model of pathogenesis. AB - Juvenile dermatomyositis (JDM), the most common pediatric inflammatory myopathy, is a systemic vasculopathy affecting young children. Epidemiology studies documenting an antecedent illness in the 3 mo before the first definite symptom (rash and/or weakness) of JDM are supported by immunologic data that suggest that the disease pathophysiology is Ag driven. The purpose of this study was to compare the gene expression profiles in muscle biopsies of four untreated DQA1*0501(+) JDM children with profiles from children with a known necrotizing myopathy (Duchenne muscular dystrophy), as well as an in vitro antiviral model (NF90), and healthy pediatric controls. Nearly half (47%) of the dysregulated genes in JDM were associated with the immune response. In particular, increased expression of IFN-alphabeta-inducible genes 6-16, myxovirus resistance protein p78, latent cytosolic transcription factor, LMP2, and TAP1 was observed. This profile is consistent with an IFN-alphabeta transcription cascade seen in the in vitro viral resistance model. The IFN-alphabeta-inducible profile was superimposed on transcription profiles reflective of myofiber necrosis and regeneration shared with Duchenne muscular dystrophy. Expressed genes were confirmed by quantitative real-time PCR (6-16), immunofluorescence (thrombospondin 4), and immunolocalization (IFN-gamma, p21). We hypothesize that these data support a model of Ag (?viral) induction of an apparent autoimmune disease based on dynamic interaction between the muscle, vascular, and immune systems in the genetically susceptible (DQA1*0501(+)) child. PMID- 11937577 TI - Depletion of collagen II-reactive T cells and blocking of B cell activation prevents collagen II-induced arthritis in DBA/1j mice. AB - Collagen II (CII)-induced arthritis in DBA/1j mice is mediated by both CII reactive T cells and anti-CII Ab-producing B cells. To determine the relative role of these processes in the development of arthritis, we specifically eliminated CII-reactive T cells by treating the mice with CII-pulsed syngeneic macrophages that had been transfected with a binary adenovirus system. These macrophages express murine Fas ligand in a doxycycline-inducible manner with autocrine suicide inhibited by concomitant expression of p35. The mice were treated i.v. with four doses of CII-APC-AdFasLp35Tet or a single dose of AdCMVsTACI (5 x 10(9) PFU), or both simultaneously, beginning 2 wk after priming with CII in CFA. Treatment with CII-APC-AdFasLp35Tet alone or in combination with a single dose of AdCMVsTACI prevented the development of CII-induced arthritis and T cell infiltration in the joint. The elimination of T cells was specific in that a normal T cell response was observed on stimulation with OVA after treatment with CII-APC-AdFasLp35Tet. Treatment with AdCMVsTACI alone prevented production of detectable levels of circulating anti-CII autoantibodies and reduced the severity of arthritis but did not prevent its development. These results indicate that the CII-reactive T cells play a crucial role in the development of CII-induced arthritis and that the anti-CII Abs act to enhance the development of CII-induced arthritis. PMID- 11937578 TI - De novo central nervous system processing of myelin antigen is required for the initiation of experimental autoimmune encephalomyelitis. AB - We demonstrate the absolute requirement for a functioning class II-restricted Ag processing pathway in the CNS for the initiation of experimental autoimmune encephalomyelitis (EAE). C57BL/6 (B6) mice deficient for the class II transactivator, which have defects in MHC class II, invariant chain (Ii), and H 2M (DM) expression, are resistant to initiation of myelin oligodendrocyte protein (MOG) peptide, MOG(35-55)-specific EAE by both priming and adoptive transfer of encephalitogenic T cells. However, class II transactivator-deficient mice can prime a suboptimal myelin-specific CD4(+) Th1 response. Further, B6 mice individually deficient for Ii and DM are also resistant to initiation of both active and adoptive EAE. Although both Ii-deficient and DM-deficient APCs can present MOG peptide to CD4(+) T cells, neither is capable of processing and presenting the encephalitogenic peptide of intact MOG protein. This phenotype is not Ag-specific, as DM- and Ii-deficient mice are also resistant to initiation of EAE by proteolipid protein peptide PLP(178-191). Remarkably, DM-deficient mice can prime a potent peripheral Th1 response to MOG(35-55), comparable to the response seen in wild-type mice, yet maintain resistance to EAE initiation. Most striking is the demonstration that T cells from MOG(35-55)-primed DM knockout mice can adoptively transfer EAE to wild-type, but not DM-deficient, mice. Together, these data demonstrate that the inability to process antigenic peptide from intact myelin protein results in resistance to EAE and that de novo processing and presentation of myelin Ags in the CNS is absolutely required for the initiation of autoimmune demyelinating disease. PMID- 11937580 TI - Genetic analysis of autoimmune sialadenitis in nonobese diabetic mice: a major susceptibility region on chromosome 1. AB - The nonobese diabetic (NOD) mouse strain provides a good study model for Sjogren's syndrome (SS). The genetic control of SS was investigated in this model using different matings, including a (NOD x C57BL/6 (B6))F(2) cross, a (NOD x NZW)F(2) cross, and ((NOD x B6) x NOD) backcross. Multiple and different loci were detected depending on parent strain combination and sex. Despite significant complexity, two main features were prominent. First, the middle region of chromosome 1 (chr.1) was detected in all crosses. Its effect was most visible in the (NOD x B6)F(2) cross and dominated over that of other loci, including those mapping on chr.8, 9, 10, and 16; the effect of these minor loci was observed only in the absence of the NOD haplotype on chr.1. Most critically, the chr.1 region was sufficient to trigger an SS-like inflammatory infiltrate of salivary glands as shown by the study of a new C57BL/6 congenic strain carrying a restricted segment derived from NOD chr.1. Second, several chromosomal regions were previously associated with NOD autoimmune phenotypes, including Iddm (chr.1, 2, 3, 9, and 17, corresponding to Idd5, Idd13, Idd3, Idd2, and Idd1, respectively), accounting for the strong linkage previously reported between insulitis and sialitis, and autoantibody production (chr.10 and 16, corresponding to Bana2 and Bah2, respectively). Interestingly, only two loci were detected in the (NOD x NZW)F(2) cross, on chr.1 in females and on chr.7 in males, probably because of the latent autoimmune predisposition of the NZW strain. Altogether these findings reflect the complexity and heterogeneity of human SS. PMID- 11937579 TI - Phenotypic and functional heterogeneity of EBV epitope-specific CD8+ T cells. AB - High frequencies of EBV-specific CD8(+) T cells have been detected during acute EBV infection, yet persistent infection inevitably results. To address this issue, we characterized the phenotype and function of epitope-specific CD8(+) T cell populations from presentation with acute through latent infection. Considerable phenotypic and functional heterogeneity within, as well as between, two different epitope-specific populations was observed over time following acute infection. B7 EBV-encoded nuclear Ag (EBNA)-3A-specific CD8(+) T cells expressed only CD45RO from acute through latent EBV infection. A2 BMLF-1-specific CD8(+) T cells expressed CD45RO during acute infection and either CD45RA or CD45RO during latent EBV infection. This difference in CD45 isoform expression between the two epitope-specific populations did not translate into differences in perforin content, the ability to produce IFN-gamma, or the ability to proliferate in response to Ag in vitro. In individuals with latent EBV infection, the frequencies of A2 BMLF-1- or B7 EBNA-3A-specific CD8(+) T cells that expressed CD45RA, CD45RO, CD62 ligand, CCR7, and perforin were stable over time. However, the expression of CD62 ligand and CCR7 was significantly higher among EBNA-3A specific CD8(+) T cells than among BMLF-1-specific CD8(+) T cells. Further work is necessary to understand how phenotypic and functional differences between EBV epitope-specific CD8(+) T cells are related to the biology of the virus and to the equilibrium between the virus and the host during persistent infection. PMID- 11937581 TI - Two distinctly HLA-associated contiguous linear epitopes uniquely expressed within the islet antigen 2 molecule are major autoantibody epitopes of the diabetes-specific tyrosine phosphatase-like protein autoantigens. AB - The related tyrosine phosphatase-like proteins islet Ag (IA)-2 and IA-2beta are autoantigens of type 1 diabetes in humans. Autoantibodies are predominantly against IA-2, and IA-2-specific epitopes are major autoantibody targets. We used the close homology of IA-2 and IA-2beta to design chimeras and mutants to identify humoral IA-2-specific epitopes. Two major IA-2 epitopes that are absent from the related autoantigens IA-2beta and IA-2Delta 13 splice variant ICA512.bdc were found contiguous to each other within IA-2 juxtamembrane amino acids 611-620 (epitope JM1) and 621-630 (epitope JM2). JM1 and JM2 are recognized by sera from 67% of patients with IA-2 Abs, and relatives of patients with type 1 diabetes having Abs to either JM epitope had a >50% risk for developing type 1 diabetes within 6 years, even in the absence of diabetes-associated HLA genotypes. Remarkably, the presence of Abs to one of these two epitopes was mutually exclusive of the other; JM2 Abs and not JM1 Abs were found in relatives with HLA DR3/4, DR4/13, or DR1/4 genotypes; and the binding of autoantibodies to the JM2 epitope, but not the JM1 epitope, markedly affected proteolysis of IA-2. This is a unique demonstration of HLA-associated B cell responses to epitopes within a single autoantigen in humans and is consistent with modification of Ag processing by specific Ab-influencing peptide presentation by HLA molecules. PMID- 11937582 TI - Antiphospholipid antibodies induce monocyte chemoattractant protein-1 in endothelial cells. AB - The presence of antiphospholipid Ab is associated with increased risk of thrombosis. The monocyte-endothelial cell interaction has been suggested to play a key role at the site of vascular injury during thrombosis. Therefore, we tested the effect of anticardiolipin Abs (aCL) on the production of monocyte chemoattractant protein-1 (MCP-1) in HUVEC. We found that monoclonal aCL as well as IgG fractions from patients with antiphospholipid syndrome (APS-IgG) could induce the production of MCP-1 in HUVEC. The ability of IgG aCL to induce MCP-1 production could be abrogated by preabsorption with cardiolipin liposomes. Simultaneous addition of either monoclonal aCL or APS-IgG with IL-1beta resulted in synergistic increase in MCP-1 production, whereas the addition of control IgG lacking aCL activity did not alter IL-1beta-induced levels of MCP-1. MCP-1 mRNA expression was also up-regulated when HUVEC were incubated with either APS-IgG or monoclonal aCL, and down-regulated by the treatment of dexamethasone. In addition, we found that serum levels of MCP-1 in 76 systemic lupus erythematosus patients correlated well with the titers of IgG aCL. Collectively, these results indicate that aCL could promote endothelial cell-monocyte cross-talk by enhancing the endothelial production of MCP-1, thereby shifting the hemostatic balance toward the prothrombotic state of APS. PMID- 11937583 TI - A role for IL-15 in driving the onset of spontaneous autoimmune thyroiditis? AB - The obese strain (OS) of chickens, which suffers from spontaneous autoimmune thyroiditis, is an excellent animal model for Hashimoto's thyroiditis and provides a unique opportunity to investigate the mechanisms underlying and driving the onset of the disease. Following recent advances in cloning chicken cytokines, we can now begin to investigate the role of cytokines in driving the lymphoid infiltration of the thyroid seen in these birds from day 7 posthatch. Using real-time quantitative RT-PCR, we characterized the expression of IFN gamma, IL-1beta, IL-2, IL-6, IL-8, IL-15, and IL-18 in thyroids from OS birds and control CB line birds, both in the embryo just before hatch (embryonic day 20) and at 3 and 5 days posthatch. All of these cytokines were up-regulated compared with levels in thyroids from CB birds, at least at some time points, with some evidence for coordination of regulation, e.g., for the proinflammatory cytokines IL-1beta and IL-8. Only IL-15 was up-regulated at all time points. IL-15 was also shown to be up-regulated in spleens of OS birds at embryonic day 20 and 5 days posthatch, suggesting that IL-15 is constitutively up-regulated in this line of birds. This could explain the general immune system hyperreactivity exhibited by OS chickens and may be a factor driving the lymphoid infiltration of the thyroid. PMID- 11937584 TI - Preferential induction of IL-10 in APC correlates with a switch from Th1 to Th2 response following infection with a low pathogenic variant of Theiler's virus. AB - Theiler's murine encephalomyelitis virus induces immune-mediated demyelination in susceptible mice after intracerebral inoculation. A naturally occurring, low pathogenic Theiler's murine encephalomyelitis virus variant showed a single amino acid change within a predominant Th epitope from lysine to arginine at position 244 of VP1. This substitution is the only one present in the entire viral capsid proteins. In this paper, we demonstrate that the majority of T cells specific for VP1(233-250) and VP2(74-86) from wild-type virus-infected mice are Th1 type and these VP1-specific cells poorly recognize the variant VP1 epitope (VP1(K244R)) containing the substituted arginine. In contrast, the Th2-type T cell population specific for these epitopes predominates in variant virus-infected mice. Immunization with UV-inactivated virus or VP1 epitope peptides could not duplicate the preferential Th1/Th2 responses following viral infection. Interestingly, the major APC populations, such as dendritic cells and macrophages, produce IL-12 on exposure to the pathogenic wild-type virus, whereas they preferentially produce IL-10 in response to the low pathogenic variant virus. Thus, such a spontaneous mutant virus may have a profoundly different capability to induce Th-type responses via selective production of cytokines involved in T cell differentiation and the consequent pathogenicity of virally induced immune-mediated inflammatory diseases. PMID- 11937585 TI - Vaccination with a Melan-A peptide selects an oligoclonal T cell population with increased functional avidity and tumor reactivity. AB - Both the underlying molecular mechanisms and the kinetics of TCR repertoire selection following vaccination against tumor Ags in humans have remained largely unexplored. To gain insight into these questions, we performed a functional and structural longitudinal analysis of the TCR of circulating CD8(+) T cells specific for the HLA-A2-restricted immunodominant epitope from the melanocyte differentiation Ag Melan-A in a melanoma patient who developed a vigorous and sustained Ag-specific T cell response following vaccination with the corresponding synthetic peptide. We observed an increase in functional avidity of Ag recognition and in tumor reactivity in the postimmune Melan-A-specific populations as compared with the preimmune blood sample. Improved Ag recognition correlated with an increase in the t(1/2) of peptide/MHC interaction with the TCR as assessed by kinetic analysis of A2/Melan-A peptide multimer staining decay. Ex vivo analysis of the clonal composition of Melan-A-specific CD8(+) T cells at different time points during vaccination revealed that the response was the result of asynchronous expansion of several distinct T cell clones. Some of these T cell clones were also identified at a metastatic tumor site. Collectively, these data show that tumor peptide-driven immune stimulation leads to the selection of high-avidity T cell clones of increased tumor reactivity that independently evolve within oligoclonal populations. PMID- 11937586 TI - Incremental value of assessment of regional wall motion for detection of multivessel coronary artery disease in exercise (201)Tl gated myocardial perfusion imaging. AB - Assessment of reversible perfusion defects in exercise (201)Tl perfusion SPECT has low sensitivity and high specificity for detection of multivessel coronary artery disease (CAD). The goal of this study was to evaluate whether worsening of left ventricular regional wall motion assessed by an automated algorithm in exercise (201)Tl electrocardiography-gated SPECT had incremental diagnostic value over perfusion data for detection of multivessel CAD. METHODS: Two hundred one patients underwent exercise (201)Tl gated SPECT. Software that automatically analyzes left ventricular function was used to assess exercise and rest regional wall motion. Regional wall motion on initial images was compared with that on rest images, that is, delayed images for patients without reinjection images and reinjection images for patients with reinjection images. The left ventricle was divided into 9 segments, with individual segments assigned to 3 coronary territories. Worsening of wall motion was defined as worsening in any segment on initial images compared with rest images. RESULTS: Of 73 patients with multivessel CAD, 20 (27.4%) had reversible perfusion defects in multiple coronary territories, 26 (35.6%) exhibited worsening of regional wall motion in multiple territories, and 37 (50.7%) had reversible perfusion defects or worsening of regional wall motion in multiple territories. The sensitivity of the combination of reversible perfusion defect and worsening of regional wall motion was significantly higher than that of reversible perfusion defect alone for detection of multivessel CAD (50.7% vs. 27.4%, P < 0.05). The specificity of the combination of reversible perfusion defect and worsening of regional wall motion for detecting multivessel CAD did not differ from that of reversible perfusion defect alone and that of worsening of regional wall motion alone (94.5% vs. 99.2% and 97.7%, respectively, P = not statistically significant). CONCLUSION: Combined assessment of worsening of left ventricular regional wall motion by exercise and perfusion data in exercise (201)Tl gated myocardial SPECT was more sensitive, with acceptable specificity, than was assessment with perfusion data alone for detection of multivessel CAD. PMID- 11937587 TI - 201Tl SPECT abnormalities, documented at rest in dilated cardiomyopathy, are related to a lower than normal myocardial thickness but not to an excess in myocardial wall stress. AB - This study was aimed at determining whether the (201)Tl SPECT abnormalities documented in patients with dilated cardiomyopathy are related to a local excess in wall stress, which might act against the diastolic perfusion of myocardium. METHODS: We included 6 healthy volunteers and 7 patients with idiopathic dilated cardiomyopathy who underwent (201)Tl SPECT at rest. On a 13-segment division of the left ventricle, indices of wall stress and tension were calculated at end diastole by applying Laplace's law, with thickness and curvature radii being determined for each segment on 2 orthogonal MRI slices. RESULTS: Among all patients, 21 analyzed segments had (201)Tl SPECT defects (D+) and 67 had none (D ). Myocardial thickness was lower in D+ (0.88 +/- 0.30 cm) than in D- (1.23 +/- 0.33 cm, P = 0.0002) or in segments from healthy volunteers (0.99 +/- 0.15 cm, P = 0.04). The index of end-diastolic wall tension was also lower in D+ (2.5 +/- 1.0 N.m(-1).mm Hg(-1)) than in D- (3.3 +/- 1.1 N.m(-1).mm Hg(-1), P = 0.02) or in segments from healthy volunteers (3.2 +/- 1.2 .m(-1).mm Hg(-1)) P = 0.04). Last, the index of end-diastolic wall stress, determined by the ratio of wall tension index to myocardial thickness, was equivalent in D+, in D-, and in segments from healthy volunteers (respectively, 3.0 +/- 1.4, 2.8 +/- 1.2, and 3.2 +/- 1.6 hN.m( 2).mm Hg(-1)). CONCLUSION: In patients with dilated cardiomyopathy, the abnormalities documented by (201)Tl SPECT at rest are related to a lower than normal wall thickness and not to an excess in wall stress or tension. Therefore, partial-volume effects are likely to induce these abnormalities, and they may be unrelated to any insufficiency of myocardial perfusion. PMID- 11937588 TI - Analysis of clinical brain SPECT data based on anatomic standardization and reference to normal data: an ROC-based comparison of visual, semiquantitative, and voxel-based methods. AB - The technique of anatomic standardization and comparison with normal templates is increasingly used in clinical brain SPECT practice and allows automated, operator independent volume-of-interest (VOI) or voxel-based analysis of whole-brain data. In 2 distinct clinical populations with severe traumatic brain injury and cognitive impairment, this study compared 3 widely available approaches that use normal templates to evaluate SPECT brain perfusion deficits. METHODS: In total, 74 subjects were studied. These included 14 patients with severe, traumatic brain injury (group 1; 10 males, 4 females; mean age +/- SD, 27.6 +/- 8.2 y) and 15 patients with cognitive impairment (group 2; 7 males, 8 females; mean age, 75.8 +/- 8.6 y). These data were compared with those from, respectively, 25 and 20 age and sex-adjusted healthy volunteers. All data were analyzed in 4 ways. Three semiquantitative statistical algorithms were used: statistical parametric mapping (SPM) using SPM99, brain registration and analysis of SPECT studies (BRASS) using a voxelwise region-growing technique, and a predefined VOI approach. These results were compared with visual analysis based on consensus reading by 3 experienced nuclear medicine physicians. Receiver operating characteristic (ROC) analysis was performed at various statistical cutoffs. Moreover, as a measure of regional agreement, relative regional agreement between methods was assessed. RESULTS: In both study groups, BRASS voxel-based analysis was most accurate, as defined by the area under the ROC curve (0.97 for group 1 and 0.96 for group 2). VOI assessment was slightly more accurate than visual consensus analysis, whereas SPM showed, overall, a lower area under the ROC curve. SPM analysis was also significantly less sensitive at thresholds corresponding to low false-positive fractions. Regional analysis showed 83%-92% agreement between all methods. CONCLUSION: Under clinical conditions, classification of brain SPECT studies can greatly be aided by anatomic standardization techniques and reference to normal data. Under the investigated circumstances, SPM was found to have a lower sensitivity than VOI or voxelwise region-growing techniques, especially at low false-positive fractions. PMID- 11937589 TI - Mapping of local renal blood flow with PET and H(2)(15)O. AB - We developed a noninvasive method for the mapping of regional renal blood flow in humans using PET and H(2)(15)O. METHODS: Fifteen subjects participated in the study, 5 with normal renal function and 10 with renal disease. The protocol used a whole-body PET scanner, intravenous bolus injection of 1,110-1,850 MBq H(2)(15)O and sequential imaging at 3 s per frame. (131)I-Iodohippuran was used to independently assess effective renal plasma flow in each subject. Hippuran clearance and renal blood flow (RBF) were measured twice, before and after treatment with probenecid, to verify that RBF is not affected. Flow analysis was based on the Kety model, according to the operational equation: C(t) = F integral C(a)(u)du - k integral C(u)du, where F is the RBF, k is the tissue-to-blood clearance rate, C is the PET concentration, and C(a) is the tracer concentration in the abdominal aorta. F and k were estimated by linear least squares on a pixel by-pixel basis to produce quantitative maps (parametric images) of RBF. The flow maps were analyzed by regions of interest (largely excluding the medulla and collecting system) for each kidney on each slice and pooled to yield mean RBF. RESULTS: In the 5 healthy subjects, mean RBF was 3.4 +/- 0.4 mL/min/g. There was no difference in flow between kidneys (t = -0.59; n = 11; P > 0.95). Before treatment with probenecid, RBF was linearly related to hippuran clearance (r(2) = 0.92). Probenecid treatment significantly reduced hippuran clearance (P < 0.003), but RBF was unchanged (P > 0.17). Compared with healthy control subjects, RBF was significantly decreased in patients with renal disease (P < 0.002). Flow maps were of good quality in all subjects, exhibiting characteristic patterns, with higher values in regions composed largely of renal cortex. CONCLUSION: Parametric mapping of RBF with PET and H(2)(15)O provides a straightforward, noninvasive method for quantitative mapping of RBF, which may prove useful in research applications and in the management of patients whose therapy alters renal tubular transport. PMID- 11937590 TI - SPECT imaging in head injury interpreted with statistical parametric mapping. AB - This study investigated regional cerebral blood flow in head-injured patients using statistical parametric mapping (SPM) to detect hypoperfusion on (99m)Tc hexamethylpropyleneamine oxime (HMPAO) SPECT scans. METHODS: Acute and follow-up SPECT and MRI scans from 61 patients who were admitted to a regional neurosurgical unit were examined. Patients had acute MRI and SPECT at 2-18 d after injury and on follow-up between 130 and 366 d after injury. Thirty-two scans from non-head-injured patients were used as a SPECT control group. The SPECT images were first aligned to the Talairach-Tournoux atlas and then analyzed statistically with SPM. RESULTS: SPECT detected more extensive abnormality than MRI in acute and follow-up stages. This effect was more pronounced on follow-up of patients with diffuse injury. Examination of a focal injury group indicated the involvement of frontal and temporal lobes and the anterior cingulate. Blood flow abnormalities persist, to a lesser extent, on follow-up scans. The diffuse group displayed low blood flow in the frontal and temporal lobes, including cingulate involvement, which persists at follow-up with additional involvement of the thalamus. CONCLUSION: SPM has a role in SPECT image interpretation because it allows better visualization than other methods of quantitative analysis of the spatial distribution of abnormalities in focal and diffuse head injury. Frontal lobe blood flow abnormality (particularly anterofrontal regions and mesiofrontal areas) is common after head injury. PMID- 11937591 TI - Clinical spectrum of asymptomatic femoral neck abnormal uptake on bone scintigraphy. AB - The purpose of this study was to assess the clinical spectrum of asymptomatic abnormal focal uptake of (99m)Tc-methylene diphosphonate (MDP) in the femoral neck. METHODS: Fifteen consecutive patients with asymptomatic abnormal focal uptake of (99m)Tc-MDP in the femoral neck were evaluated. Two patients had bilateral abnormal focal uptake. The patient's history, clinical findings, and plain hip radiograph were obtained in all cases. Scintigraphic, radiographic and clinical findings were correlated. RESULTS: Eight of 17 (47%) femoral necks showed a definite herniation pit on radiography, 6 patients (35%) had normal radiographic findings, 1 patient had a bone island, 1 patient had a bone island and a herniation pit, and 1 patient had a subtle lesion suggestive of a herniation pit on radiography. All patients remained asymptomatic for at least a 10-mo follow-up period. CONCLUSION: A herniation pit is the most common finding among asymptomatic abnormal femoral neck focal uptake. This condition should be distinguished from a wide variety of disorders associated with increased focal abnormal uptake of (99m)Tc-MDP in the femoral neck. PMID- 11937592 TI - Beta-agonist aerosol distribution in respiratory syncytial virus bronchiolitis in infants. AB - Bronchodilator aerosols are frequently administered to infants with bronchiolitis but with little success. The efficacy of aerosol treatments depends mainly on adequate targeting of the aerosol particles to the inflamed airways. This study evaluated the lower respiratory tract distribution characteristics of nebulized bronchodilators in infants with acute bronchiolitis. METHODS: Twelve infants (mean age +/- SD, 8 mo +/- 4 mo) who were admitted for acute respiratory syncytial virus bronchiolitis were treated with (99m)Tc-albuterol aerosol. Gamma scintigraphy was used to assess total body and lung deposition as well as pulmonary distribution of the medication. RESULTS: Of the total 6-min nebulized dose (i.e., drug aerosol dose leaving the nebulizer [not the nebulizer charge]), 1.5% +/- 0.7% reached the right lung, with only approximately one third of that (0.6%) penetrating to the peripheral lung zone. There was 7.8% +/- 4.9% deposition in the upper respiratory and gastrointestinal tracts and 10%-12% remained on the face. No correlation was found between any of the deposition indices and the clinical response data or any of the demographic parameters (e.g., height, weight, body surface area, or clinical score). CONCLUSION: Poor total aerosol deposition in infants may be related as much to their small conducting airways as to the disease state. There is considerable room for improvement in aerosol delivery in this age group, with greater emphasis on targeting narrowed peripheral airways with superfine aerosols. PMID- 11937593 TI - The clinical impact of (18)F-FDG PET in patients with suspected or confirmed recurrence of colorectal cancer: a prospective study. AB - This prospective study aimed to confirm, in a clinical setting, the benefits suggested by earlier retrospective studies of (18)F-FDG PET scanning for the evaluation of patients with suspected recurrence of colorectal cancer. METHODS: The referring oncologist was asked to prospectively assign a treatment plan for 102 consecutive patients being evaluated by (18)F-FDG PET for suspected or confirmed recurrence of colorectal cancer and without evidence of unresectable disease on conventional staging investigations, including CT. This treatment plan was then compared with that based on incremental information supplied by PET. Management changes were validated by follow-up. RESULTS: For 6 patients, the oncologist would not commit to a management plan without access to PET information, and for all these patients, PET correctly guided management. Of the remaining 96 patients, the management plan for 54 (56%) was altered as a direct result of unexpected PET findings. Thus, PET directly influenced management in 60 (59%) of 102 patients. The discrepant PET results could be validated in 57 patients and were correct for both the presence and the extent of malignant disease in 52 (91%) of these patients but were false-positive in 1 patient because of a pelvic abscess and underestimated the extent of metastatic disease in 4 (7%). Relapse was confirmed in 49 (98%) of 50 evaluable patients with positive PET findings. Significantly, planned surgery was abandoned in 26 (60%) of 43 patients because of incremental PET findings. Of the 42 patients for whom management was not changed by PET findings, false-negative PET findings were documented for 5 (4 with metastases < 1 cm), and the PET findings for 1 were presumed to be false-positive because of sarcoidosis. CONCLUSION: This prospective study confirms the high impact, suggested by previous retrospective analyses, of (18)F-FDG PET on management of patients with suspected recurrent colorectal cancer. The major benefit of PET is avoidance of inappropriate local therapies by documentation of widespread disease. PMID- 11937594 TI - Blood flow and metabolism in locally advanced breast cancer: relationship to response to therapy. AB - Locally advanced breast cancer (LABC) is commonly treated with neoadjuvant chemotherapy followed by definitive surgery. The factors influencing the response of LABC to presurgical chemotherapy are incompletely understood. To characterize in vivo tumor biology in patients with LABC, we measured pretherapy blood flow and glucose metabolism in LABC, compared measurements with clinical and pathologic parameters, and examined blood flow and response to subsequent neoadjuvant chemotherapy. METHODS: Thirty-seven patients with newly diagnosed LABC underwent (18)F-FDG and (15)O-water PET imaging. Thirty-one of these patients underwent neoadjuvant chemotherapy, and response was evaluated by serial measurements of tumor size and pathologic examination after definitive surgery after chemotherapy. Tumor metabolism was estimated from graphic analysis of dynamic (18)F-FDG studies and was expressed as the metabolic rate of (18)F-FDG (MRFDG). Blood flow was estimated from dynamic images after bolus (15)O-water injection using a 1-compartment model. Tumor blood flow and metabolism were compared with clinical and pathologic parameters and with response to chemotherapy. RESULTS: Both blood flow and metabolism were significantly higher in tumor than in normal breast. Tumor blood flow and metabolism were correlated but highly variable. There were weak associations of metabolism with patient age and tumor grade and of blood flow with estrogen receptor status. There was a statistically significant trend for patients with a high MRFDG to have a poorer response to therapy (P = 0.001). Response was not significantly correlated with any other parameters. A low ratio of MRFDG to blood flow was the best predictor of macroscopic complete response (CR) (P = 0.02 vs. non-CR). Preliminary analysis of patient follow-up showed the ratio of MRFDG to blood flow to also be predictive of disease-free survival. CONCLUSION: Despite uniformly large tumor size, blood flow and metabolism in LABC are highly variable. High glucose metabolism predicts a poor response to neoadjuvant chemotherapy, and low MRFDG relative to blood flow is a predictor of CR. Further work is needed to elucidate the biologic mechanisms underlying these findings. PMID- 11937595 TI - The role of quantitative (18)F-FDG PET studies for the differentiation of malignant and benign bone lesions. AB - The role of quantitative (18)F-FDG PET studies for the differentiation of benign and malignant bone lesions is still an open question. METHODS: Our evaluation included 83 patients with 37 histologically proven malignancies and 46 benign lesions. Thirty-five of the 46 benign lesions were histologically confirmed. The (18)F-FDG studies were accomplished as a dynamic series for 60 min. Evaluation of the (18)F-FDG kinetics was performed using the following parameters: standardized uptake value (SUV), global influx (Ki), computation of the transport constants K1 k4 with consideration of the distribution volume (VB) according to a 2-tissue compartment model, fractal dimension based on the box-counting procedure (parameter for the inhomogeneity of the tumors). RESULTS: The mean SUV, the vascular fraction VB, K1, and k3 were higher in malignant tumors compared with benign lesions (t test; P < 0.05). Although the (18)F-FDG SUV was helpful to differentiate benign and malignant tumors, there was some overlap, which limited the diagnostic accuracy. On the basis of the discriminant analysis, the SUV alone showed a sensitivity of only 54.05%, a specificity of 91.30%, and a diagnostic accuracy of 74.70%. The fractal dimension was superior and showed a sensitivity of 71.88%, a specificity of 81.58%, and an accuracy of 77.14%. The combination of SUV, fractal dimension, VB, K1-k4, and Ki revealed the best results with a sensitivity of 75.86%, a specificity of 97.22%, and an accuracy of 87.69%. Bayesian analysis showed true-positive results at the level of 0.8 for a low prevalence of disease (0.235) if the full kinetic data were used in the evaluation. CONCLUSION: (18)F-FDG PET has a high specificity for the exclusion of a malignant bone tumor. Evaluation of the full (18)F-FDG kinetics and the application of discriminant analysis are required and can be used prospectively to classify a bone lesion as malignant or benign. PMID- 11937596 TI - 99mTc-sestamibi imaging in the assessment of toremifene as a modulator of multidrug resistance in patients with breast cancer. AB - Multidrug resistance (MDR) due to expression of a membrane-associated permeability glycoprotein (P-glycoprotein [Pgp]) prevents successful cytotoxic chemotherapy for breast cancer. Identification of MDR would facilitate selection of chemotherapy regimens and MDR modulators. This study aimed to evaluate (99m)Tc sestamibi imaging for predicting overexpression of Pgp in primary breast cancer and to measure the efficacy of toremifene, the MDR modulator, in vivo. METHODS: Twenty patients with untreated breast cancer had (99m)Tc-sestamibi imaging 20 and 120 min after tracer injection before and after a 3-d course of toremifene (780 mg/d). Tumor samples were obtained during surgery for correlation of imaging and Pgp immunohistochemistry. RESULTS: Sixteen of 20 tumors were visualized with sestamibi. Before toremifene, there was a significant inverse correlation (Spearman rank correlation coefficient [R(S)]) between staining intensity, based on the anti-Pgp monoclonal antibodies C494 and C219, and the tumor-to-background ratio (T/B) at 120 min (R(S) = -0.85; P < 0.001 and R(S) = -0.71; P < 0.001, respectively). However, the correlation between the T/B and immunohistochemistry at 20 min was significant only for C494 (R(S) = -0.57; P < 0.01). Similarly, before toremifene, there was an inverse correlation between staining intensity and the change in the T/B between 20 and 120 min (R(S) = -0.77; P < 0.001 and 0.75; P < 0.001 for C494 and C219). After toremifene, an inverse correlation between staining intensity and the T/B was seen only at 120 min and only with C494 (R(S) = -0.68; P < 0.01). However, the change in the T/B between 20 and 120 min correlated significantly with staining intensity for C494 and C219 (R(S) = 0.68; P < 0.01 and -0.7; P < 0.01 for C494 and C219, respectively). Toremifene did not significantly alter the overall T/B at either 20 or 120 min when data were compared before and after toremifene. Nevertheless, at 120 min, 8 of 8 tumors with low Pgp expression showed reduced uptake after toremifene, whereas 5 of 6 tumors with strong expression showed increased uptake (P < 0.003). Moreover, there was a significant correlation between the change in the T/B and staining intensity with C494 (R(S) = 0.59; P < 0.05) and C219 (R(S) = 0.56; P < 0.05) at 120 min but not at 20 min. CONCLUSION: (99m)Tc-Sestamibi accumulation in breast cancer correlates with Pgp expression. Toremifene has a dual effect on this accumulation, increasing it through an inhibitory effect on Pgp while at the same time reducing it by a direct competition with sestamibi. The latter implies that in response to Pgp modulation the efflux of various agents may be affected differently. PMID- 11937597 TI - What can we expect from MDR breast cancer imaging with sestamibi? PMID- 11937598 TI - Effects of carvedilol on cardiac function and cardiac adrenergic neuronal damage in rats with dilated cardiomyopathy. AB - Metaiodobenzylguanidine (MIBG) is a reliable marker for the detection of cardiac adrenergic neuronal damage in heart failure. The cardioprotective properties of carvedilol, a vasodilating beta-adrenoceptor-blocking agent, were studied in a rat model of dilated cardiomyopathy after autoimmune myocarditis. METHODS: Twenty eight days after immunization, surviving rats (41/55, or 75%) were divided into 2 groups treated with carvedilol, 2 mg/kg/d (group C, n = 19), or vehicle alone (0.5% methylcellulose, group V, n = 22). After oral administration for 2 mo, heart weight, heart rate, left ventricular end-diastolic pressure (LVEDP), and myocardial fibrosis were measured and compared with those in untreated rats (group N, n = 19). Myocardial uptake of (125)I-MIBG (differential absorption ratio) in the left ventricle was measured by autoradiography at 10, 30, or 240 min after tracer injection. RESULTS: Four (18%) of 22 rats in group V died between days 28 and 84 after immunization. None of the rats in group C or N died. Heart weight, heart rate, LVEDP, and area of myocardial fibrosis in group C (1.14 +/- 0.04 g, 345 +/- 16 beats per minute, 7.6 +/- 1.5 mm Hg, and 12% +/- 1%) were significantly lower than those in group V (1.34 +/- 0.04 g, 389 +/- 10 beats per minute, 12.3 +/- 1.3 mm Hg, and 31% +/- 2%). Although the differential absorption ratio was lower at all time points in group V than in group N, uptake after treatment increased in group C, compared with group V, at 10 min (12.5 +/- 1.0 vs. 7.6 +/- 0.8, not significant), 30 min (10.1 +/- 1.1 vs. 6.3 +/- 0.9, not significant), and 240 min (6.5 +/- 0.5 vs. 2.5 +/- 0.2, P < 0.05). The late washout ratio from myocardial radioactivity between 30 and 240 min in group C was lower than that in group V (36% vs. 60%). CONCLUSION: These observations indicated that carvedilol has beneficial effects and protects cardiac adrenergic neurons in dilated cardiomyopathy. PMID- 11937599 TI - Myocardial adrenergic dysinnervation in dilated cardiomyopathy: cornerstone or epiphenomenon? PMID- 11937600 TI - Myocardial contractile reserve and perfusion defect severity with rest and stress dobutamine (99m)Tc-sestamibi SPECT in canine stunning and subendocardial infarction. AB - Myocardial contractile reserve and resting perfusion scintigraphy provide independent information to assess myocardial viability. The purpose of this study was to simultaneously evaluate both with (99m)Tc-sestamibi SPECT and low-dose dobutamine in canine stunning and subendocardial infarction (SEMI). METHODS: Eighteen dogs were included in the study: 7 normal, 7 stunned, and 4 with SEMI. Closed-chest stunning and SEMI were produced by angioplasty balloon occlusion of the left anterior descending artery (20 and 90 min, respectively). Subsequent radiolabeled mircospheres confirmed reflow, and (99m)Tc-sestamibi was then administered at rest. Gated SPECT and MRI tagging were performed at rest and during low-dose dobutamine infusion (5 microg/kg/min). SPECT systolic wall thickening index (SWI) and MRI radial strain quantified myocardial contraction. Postmortem 2,3,5-triphenyltetrazolium chloride staining quantified SEMI. RESULTS: Defect severity by SPECT in the anterior wall was mild and was not statistically different for the stunned versus SEMI groups (P = not significant). At rest, anterior wall SPECT SWI was significantly higher in the normal versus stunned groups (21.1 +/- 3.1 vs. 10.1 +/- 9.0; P = 0.0002) and the normal versus SEMI groups (21.1 +/- 3.1 vs. 2.6 +/- 6.0; P = 0.000002). With low-dose dobutamine, SWI increased significantly compared with rest for the stunned group (29.1 +/- 10.4 vs. 10.1 +/- 9.0; P = 0.000007) but did not increase significantly for the SEMI group (11.0 +/- 11.3 vs. 2.6 +/- 6.0; P = 0.09); SWI during low-dose dobutamine infusion for the stunned group was comparable to that for the normal group (29.1 +/- 10.4 vs. 28.2 +/- 7.0; P = 0.80). SWI also showed correlation with MRI radial strain (r = 0.42; P = 0.00015). CONCLUSION: Defect severity for stunned myocardium and SEMI was mild and was not significantly different. Contractile reserve was significantly different in stunned myocardium and SEMI. (99m)Tc-Sestamibi SPECT at rest and with low-dose dobutamine is a promising new technique to simultaneously assess myocardial perfusion and contractile reserve. PMID- 11937601 TI - Effect of endomyocardial laser channels on regional innervation shown with (125)I MIBG and autoradiography. AB - The aim of this study was to map regional innervation against regional flow early after laser channel placement using autoradiography in a porcine model. METHODS: Four juvenile male swine underwent left ventricular mapping using a catheter based mapping system and laser treatment with 20-30 channels to the mid and distal anterior wall of the left ventricle. Three days later animals were injected with 37 MBq (125)I-metaiodobenzylguanidine (MIBG) followed in 3 h with 1,110 MBq (99m)Tc-sestamibi; 1 h later the animals were killed. Hearts were removed, perfusion fixed, and sliced into 1-cm slices. The slices best showing laser holes were selected, and circumferential sections were taken for autoradiography and hematoxylin-eosin staining. Phosphor screens were exposed for (99m)Tc and (125)I, and images were processed. The MIBG image was subtracted from the methoxyisobutylisonitrile (MIBI) image and vice versa, and color tables were applied to the difference images and overlaid on the perfusion images. Quantitative analysis of the light image data was also performed. RESULTS: Thirty three sections from the last 3 experiments were analyzed. Acoustic damage from 30 laser channels was identified from the hematoxylin-eosin sections. Reduced MIBG relative to regional flow was seen in surrounding tissue corresponding to only 1 channel. There was no statistically significant difference in light units expressed as (MIBG - MIBI)/maximal MIBG value between laser channels and unmarked myocardial map regions. The regions identified from the color table on the map as low MIBG relative to MIBI were significantly lower than remaining laser channels and remaining myocardium. Mean light units for the regions with high MIBG relative to MIBI were significantly higher than the remaining laser channels and remaining myocardium. CONCLUSION: Using a high-resolution technique correlated with microscopic pathology in an animal model, there is negligible regional denervation 3 d after placement of endomyocardial laser channels. PMID- 11937602 TI - Myocardial blood flow, metabolism, and inotropic reserve in dogs with dysfunctional noninfarcted collateral-dependent myocardium. AB - There is intense controversy as to the mechanisms underlying chronic but reversible left ventricular (LV) ischemic dysfunction. The aim of this study was to investigate the physiology underlying this condition in a canine model of noninfarcted collateral-dependent myocardium. METHODS: Six mongrel dogs were instrumented with ameroid constrictors on the left circumflex and right coronary arteries and a partial occluder on the left anterior descending coronary artery. The animals were followed up for 6 mo. Every 6 wk, measurements of regional wall thickening (M-mode echo), myocardial blood flow ((13)N-ammonia PET), oxygen consumption ((11)C-acetate PET), and glucose uptake ((18)F-FDG PET) were obtained. After 6 mo, myocardial blood flow reserve (during adenosine infusion) and regional contractile reserve (during infusion of a low dose of dobutamine) were also investigated. RESULTS: Following ameroid implantation, regional thickening decreased in the posterior wall (to 34% +/- 13% of baseline; P < 0.001) but not in the septum. Resting myocardial blood flow (56 +/- 10 vs. 58 +/- 15 mL.[min.100 g](-1)), myocardial oxygen consumption (21 +/- 3 vs. 22 +/- 3 J.[beat.100 g](-1)), and insulin-stimulated glucose uptake (39 +/- 8 vs. 42 +/- 11 micromol.[min.100 g](-1)) were similar among dysfunctional and normal segments. Myocardial blood flow reserve was blunted in dysfunctional versus normal segments (3.7 +/- 0.5 vs. 5.2 +/- 1.5; P = 0.06). With dobutamine, wall thickening (to 69% +/- 8% and 77% +/- 11%, respectively) and oxygen consumption (to 36 +/- 5 and 39 +/- 5 J.[beat.100 g](-1), respectively) improved to the same extent in both segments. As a consequence, mechanical efficiency decreased in septal but remained unchanged in posterior segments during infusion of dobutamine. Biopsy specimens from both walls were free from any morphological alterations. CONCLUSION: Our data indicate that ameroid occlusion in dogs induces sustained reduction in regional contraction, which occurs despite normal levels of transmural blood flow and recruitable inotropic reserve. Since myocardial perfusion reserve was blunted, such perfusion-contraction mismatch could reflect repetitive stunning. PMID- 11937604 TI - External and internal dose to individuals after (131)I outpatient therapy. PMID- 11937603 TI - (201)Tl and (99m)Tc-MIBI retention in an isolated heart model of low-flow ischemia and stunning: evidence of negligible impact of myocyte metabolism on tracer kinetics. AB - It is not known whether cellular metabolic disorders play a role in the decreased tracer uptake that is documented by conventional SPECT during low-flow ischemia or stunning. This study sought to determine the impact of low-flow ischemia and stunning on the kinetics of (201)Tl and MIBI across the plasma membrane of myocytes. METHODS: The global myocardial retention (Rf) of (201)Tl and MIBI was determined in isolated working hearts from rabbits, perfused with red blood cell enhanced solution. Experiments were performed in normoxia, with physiological values of coronary flow (N; n = 16); in low-flow ischemia, with a >50% reduction of coronary flow and a > or =20-mm Hg fall in systolic left ventricle pressure (L; n = 15); and in stunning, with 15 min of acute ischemia followed by reperfusion (S; n = 15). Concentration ratios across the plasma membrane of myocytes were also determined for both tracers and expressed as Ci/Cc, where Ci is interstitial activity determined with microdialysis, and Cc is activity from cellular space determined from Rf and Ci values. RESULTS: There was a slight increase in average values of Ci/Cc in ischemia, but not in stunning, for (201)Tl (L, 0.011 +/- 0.006 vs. N, 0.006 +/- 0.004 [P < 0.05]; S, 0.007 +/- 0.004 vs. N [not significant]) and for MIBI (L, 0.011 +/- 0.008 vs. N, 0.005 +/- 0.004 [P < 0.05]; S, 0.005 +/- 0.003 vs. N [not significant]). Moreover, ischemia and stunning had no deleterious effects on the average values of global myocardial retention for (201)Tl (L, 0.63 +/- 0.09 vs. N, 0.50 +/- 0.14 [P < 0.05]; S, 0.59 +/- 0.10 vs. N [P < 0.05]) or for MIBI (L, 0.45 +/- 0.10 vs. N, 0.31 +/- 0.09 [P < 0.05]; S, 0.41 +/- 0.12 vs. N [P < 0.05]). In fact, these values were significantly enhanced in the 2 situations. CONCLUSION: The kinetics of (201)Tl and MIBI across the plasma membrane of myocytes were affected only poorly by low flow ischemia and not at all by stunning, without any deleterious effects on myocardial retention of both tracers. During low-flow ischemia or stunning, therefore, the information provided by (201)Tl or MIBI SPECT is expected to depend on myocardial perfusion but not on cellular metabolic disorders. PMID- 11937606 TI - New York City, 2001: reaction and response. PMID- 11937607 TI - Public health preparedness for disaster: perspective from Washington, DC. PMID- 11937608 TI - Of butterflies, madmen, and urban health: perspective from Canada. PMID- 11937609 TI - Can cities prepare for terrorism?: Perspective from the United Kingdom. PMID- 11937610 TI - Public health and the media: the challenge now faced by bioterrorism. PMID- 11937612 TI - Science and politics, or on the irony of the term political science. PMID- 11937614 TI - The epidemiology of firearm suicide in the United States. AB - CONTEXT: Little attention has been given to the role of firearms in suicide. In 1998, firearms were the leading method of committing suicide for both men and women, responsible for three times the number of suicides compared to the next leading method. Understanding the epidemiology of firearm suicide will increase awareness of firearm suicide as a major public health problem. RESULTS: Rates of firearm suicide have changed little over the past two decades and have consistently exceeded rates of firearm homicide. The firearm suicide rate among men is approximately six times that of women. While firearm suicide rates are highest among the elderly, the majority (66%) of firearm suicides are among persons under 55 years of age. Firearm suicide rates among women of all ages have dropped modestly, while rates among elderly men have risen considerably. Whites have roughly twice the rate of firearm suicide as do blacks and other race/ethnicity groups. Individual-level empirical studies have consistently indicated that keeping firearms in the home is associated with an increased risk of suicide. CONCLUSIONS: For suicide prevention to be effective, the availability and use of firearms in suicides must be addressed. PMID- 11937613 TI - Firearm availability and suicide, homicide, and unintentional firearm deaths among women. AB - CONTEXT: In the United States, more than 45,000 women died from gun violence over the last decade. OBJECTIVE: To determine whether measures of firearm availability are related to rates of suicide, homicide, and unintentional firearm deaths among women in the United States. DESIGN: Pooled cross-sectional time series data on suicide, homicide, and unintentional firearm deaths (1988-1997) were used to estimate the association between the rate of violent death among women and four proxies of firearm availability. Two proxies came from survey reports of household firearm ownership rates; two were derived from mortality statistics. SETTING: United States, 1988-1997. RESULTS: The increased rate of suicide and homicide in states with high gun levels was accounted for primarily by significantly elevated firearm suicide and firearm homicide rates. Unintentional firearm death rates were also increased in states with more guns. At the regional level, qualitatively similar results were obtained. CONCLUSION: Between 1988 and 1997, the suicide, homicide, and unintentional firearm death rates among women were disproportionately higher in states where guns were more prevalent. The elevated rates of violent death in states with more guns was not entirely explained by a state's poverty or urbanization and was driven primarily by lethal firearm violence, not by lethal nonfirearm violence. PMID- 11937615 TI - Unintentional gun injuries, firearm design, and prevention: what we know, what we need to know, and what can be done. AB - The public health community has long recognized unintentional gun injuries as a public health issue. In 1998 in the United States, 866 people died from unintentional gunshot wounds, resulting in a crude death rate of 0.32 per 100,000. Unintentional gun deaths have been declining since at least 1920, yet the reasons for this downward trend are not understood. Possible explanations, such as changes in gun ownership and demography, changes in access to guns among population subgroups, safety practices, and artifactual influences are discussed. Intervention strategies for reducing the risk of unintentional gun injury are also discussed. PMID- 11937616 TI - How delinquent youths acquire guns: initial versus most recent gun acquisitions. AB - BACKGROUND: Access to firearms among delinquent youths poses significant risks to community safety. The purpose of the study was to describe how a group of criminally involved youths obtained guns. METHODS: Youths were randomly selected from a juvenile justice facility to participate in a semistructured, anonymous interview. Transcripts were coded and analyzed with the aid of textual analysis software. RESULTS: Of the 45 participants, 30 had acquired at least 1 gun prior to their most recent incarceration, and 22 had acquired multiple guns. About half of the first gun acquisitions were gifts or finds. The first guns youths acquired were usually obtained from friends or family. The most recent acquisitions were often new, high-caliber guns, and they came from acquaintances or drug addicts. New guns often came from high-volume traffickers. Gun acquisitions from strangers or through "straw purchases" were rare. Though few obtained guns directly through theft, some youths believed their supplier had stolen guns. Youths rarely left their community to obtain a gun. CONCLUSIONS: Guns were readily available to this sample of criminally involved youths through their social networks. Efforts to curtail high-volume, illegal gun traffickers and to recover discarded guns from areas in which illicit drug sales take place could potentially reduce gun availability to high-risk youth. PMID- 11937617 TI - Drugs and firearm deaths in New York City, 1990-1998. AB - Firearm deaths remain among the leading causes of mortality in the United States. Changing law enforcement activities, incarceration, drug use, and socioeconomic conditions may have played roles in the declining rates of firearm deaths during the 1990s. Using records from the Office of the Chief Medical Examiner, we analyzed the role of drugs in firearm deaths in New York City between 1990 and 1998. Positive drug toxicology was present in over half of all firearm death victims during this time. Cocaine, cannabis, opiates, and alcohol accounted for almost all of these deaths with drug-positive toxicology. There were decreases in cocaine- and alcohol-positive toxicology for firearm deaths in New York City starting in the early 1990s; there was a more gradual decrease in heroin-positive toxicology for firearm deaths. Cannabis-positive toxicology for firearm deaths increased in the early part of the 1990s and then decreased starting in the mid 1990s. Although the disparities between minority and white firearm death rates narrowed during this time, minorities remained about three times more likely to be victims of fatal firearm violence than whites in 1998. The highest firearm death rates were among African American and Latino male decedents, with a larger proportion of Latinos testing cocaine or opiate positive, while a larger proportion of African Americans tested cannabis positive. These results suggest a complex role of drugs in firearm-related deaths. PMID- 11937618 TI - Traffic injury data, policy, and public health: lessons from Boston Chinatown. AB - We note that long-standing land use and transportation policy are critical factors in creating traffic conditions and will have to play a role in reducing injuries. We present the historical progression of events that created current traffic conditions in Boston Chinatown and an analysis of traffic-related injuries at the community level for the years 1996-1998. Injuries were found to be as likely on weekends as on weekdays and frequently occurred late at night. Nighttime occupant injuries were found to be more likely on Friday, Saturday, and Sunday nights (relative risk = 2.26; confidence interval = 1.35-3.78, P =.0014). Injuries varied significantly by location for occupant (P = <.001) and for pedestrian injuries (P =.039). There were no peaks of injuries at traditionally defined commuter hours, which have been the standard time for assessing "worst case" traffic impacts by developers and government agencies. There was, however, a strong association between injuries and vehicle volume at 9 intersections with simple configurations for AM and PM commuter hours (R(2) = 0.589, P =.010), resulting in a calculated increase of 3-5 injuries per year for each increase of 1,000 vehicles. There was no such association at 10 intersections with complex configurations (R(2) = 0.104, P =.397). The 24-hour weekend patterns of vehicle volumes showed that traffic abated only between 3 and 7 AM, and the patterns appeared qualitatively to mirror the 24-hour pattern of injuries, suggesting that they were also indicative of injury risk. We suggest that there is a need for both long-term changes in policy and more immediate interventions. We also conclude that researchers should be cautious about assuming that traffic patterns conform to naive expectations such as rush hour peaks. PMID- 11937619 TI - Regional variation in drug purchase opportunity among youths in the United States, 1996-1997. AB - This study was designed to examine geographic variation in illegal drug purchase opportunity among young people living in the United States; there was a subfocus on age, sex, and urban/rural residence. Data from the 1996-1997 National Household Surveys on Drug Abuse were analyzed; the nationally representative sample of community residents included 21,531 participants aged 12-24 years old. Respondents were asked if someone had approached them to sell them an illegal drug during the past 30 days. To protect respondents' confidentiality, there is no finegrained geographical coding of data in the National Household Surveys on Drug Abuse public use data files, but nine geographical divisional indicators are provided (i.e., West North Central, New England, etc.). Results indicated males were an estimated 1.8 times more likely than females to have had a recent illicit drug purchase opportunity, and urban residents were 1.5 times more likely than rural residents to have had a recent drug purchase opportunity. As for geographic divisions, the Pacific division surpassed all other divisions: Its residents were 1.5 times more likely to have recent drug purchase opportunities than the West North Central division (used here as a reference category). After controlling statistically for age, sex, and urban/rural residence, residence in four divisions was found to be associated with greater likelihood of an illicit drug purchase opportunity. The observed patterns of drug purchase opportunity add new features to our understanding of illicit drug involvement across the United States. PMID- 11937620 TI - Characteristics of injection drug users who utilize tuberculosis services at sites of the Baltimore city needle exchange program. AB - OBJECTIVE: To describe characteristics of needle-exchange program (NEP) participants who utilized tuberculosis services from an NEP site. METHODS: Between June 1998 and May 1999, tuberculosis services were advertised and offered to Baltimore, Maryland, NEP participants. Demographic and tuberculosis-specific data were collected on participants who self-selected into services. Analyses were based on being tuberculin skin tested, returning for a skin test reading, and testing tuberculin positive. RESULTS: Among 691 contacts with NEP participants, this service performed 296 tuberculin tests, with an 84% return rate for skin test reading. Participants were 32% female, 87% African American, and 11% employed. Higher frequency of NEP visits was positively associated with requesting tuberculosis services and returning for skin test reading. Among those who returned for skin test reading, longer smoking duration and problems getting food in the past year due to a lack of money were associated with a positive test. CONCLUSIONS: Utilization of a tuberculosis service and high return rates can be achieved among NEP participants without formal recruitment strategies. Frequent exchange appears to facilitate return visits for NEP-based tuberculosis screening, which may imply accessibility for frequent exchangers. More extensive health services at sites of the Baltimore NEP appear to be warranted, with particular attention paid to effectiveness for frequent exchangers. PMID- 11937621 TI - Selection effect of needle exchange in Anchorage, Alaska. AB - Participation bias (selection bias) may be a problem in studies that attempt to evaluate the effects of needle-exchange programs (NEPs). The present study looked at only those injection drug users (IDUs) who were randomly placed in the needle exchange condition in a two-arm randomized clinical trial of needle exchange. Time to follow-up between the experimental NEP condition (n = 296; median = 261 days) and pharmacy sales condition (n = 304; median = 256 days) was not statistically different [chi(2) (1, N = 600) = 0.42, P =.52]. Within the NEP condition, a similar analysis comparing time to follow-up between IDUs who used the NEP (n = 65; median = 199 days) and those who refrained from using the NEP (n = 231; median = 286 days) was highly significant, chi(2) (1, n = 296) = 17.3, P=.0001. The final logistic regression model predicting use of the NEP was the log 10 transformation of the number of times injected heroin in the last 30 days (odds ratio [OR ]=4.9, confidence interval [CI ]2.8, 8.9), ever injected amphetamine in the last 30 days (OR =4.9, CI 1.09, 22.5), and ever shared injection equipment in the last 30 days (OR =2.9, CI 1.5, 5.5). Within the NEP condition, follow-up rates differed between those who used the NEP and those who did not use it. Of drug users randomly assigned to an NEP, the ones who actually used the NEP had higher levels of drug use. Predictors of who used the NEP were consistent with the studies of Hagen et al. This study demonstrates that selection bias occurs among IDUs who use NEPs. PMID- 11937622 TI - Trends in the AIDS epidemic among New York City's injection drug users: localized or citywide? AB - The New York City injection drug user acquired immunodeficiency syndrome (IDU AIDS) epidemic accounts for almost one quarter of AIDS cases in IDUs in the United States. Recent studies have reported declines in seroprevalence and risk behaviors among IDUs in New York City during the 1990s. These trends, however, are based on studies primarily conducted in the city's central borough of Manhattan. This article analyzes data from all five boroughs of New York City to examine trends over phases of the epidemic and to determine the level of prevention services available; an exploratory qualitative study was also conducted to assess access to prevention services and injection practices in areas in the "outer boroughs." Findings indicated that (1) borough differences in services and behaviors existed from early in the epidemic; (2) services have been concentrated in Manhattan; and (3) declines in seroprevalence were greatest among Manhattan-recruited IDUs. Enhancing access to services for IDUs in the boroughs outside Manhattan may be needed to continue the positive trends in all areas of New York City. PMID- 11937623 TI - The effect of ethnicity and maternal birthplace on small-for-gestational-age deliveries to HIV-infected women. AB - OBJECTIVE: To examine the relative role of ethnicity and maternal birthplace on small-for-gestational-age (SGA) deliveries of a cohort of mothers in New York who were infected with human immunodeficiency virus. METHODS: Medicaid claims and linked vital statistics records were examined for 2,525 singleton deliveries to HIV-infected women from 1993 through 1996. We estimated adjusted odds ratios (AORs) and 95% confidence intervals (CIs) of SGA delivery associated with ethnicity (i.e., white, white-Latina, black, and black-Latina) and maternal birthplace (i.e., native US/Puerto Rican vs. foreign born) in a series of multivariate regression models to which we sequentially added demographic, health services, and lifestyle factors (i.e., alcohol, tobacco, and illicit drug use). RESULTS: Of the deliveries, 10% were SGA. The odds of SGA infants for black and white women did not differ by maternal birthplace. Foreign-born white-Latinas and black-Latinas had lower unadjusted odds of a SGA delivery than their US-born counterparts (OR 0.29, CI 0.14, 0.61 and OR 0.22, CI 0.07, 0.71, respectively). After adjustment for maternal lifestyle characteristics, the odds of SGA delivery were 0.50 (CI 0.23, 1.09) for white-Latina mothers and 0.60 (CI 0.17, 2.08) for black-Latina mothers. CONCLUSIONS: SGA outcomes did not differ by maternal birthplace for black and white women. Differences in lifestyle factors appear to contribute to lower odds of SGA delivery for foreign-born versus US-born white- and black-Latina HIV-infected women. PMID- 11937624 TI - Ku86 autoantigen related protein-1 transcription initiates from a CpG island and is induced by p53 through a nearby p53 response element. AB - The human Ku86 gene and an isoform, KARP-1 (Ku86 autoantigen related protein-1), encode overlapping, but differentially regulated, transcripts. Ku86 is constitutively transcribed at high levels and, although it plays a seminal role in DNA double-strand break repair, its expression is not induced by DNA damage. KARP-1, in contrast, is expressed constitutively only at low levels and its expression is induced by DNA damage in a p53-dependent fashion. The regulatory elements promoting KARP-1 gene expression and p53 responsiveness, however, were unknown. Here, we report that a strong DNase I hypersensitive site (DHS) resides approximately 25 kb upstream from the Ku86 promoter. This DHS is encompassed by a hypomethylated CpG island. Reporter assays demonstrated that this region corresponded to a promoter(s), which promoted transcription of peroxisomal trans 2-enoyl CoA reductase in the centromeric direction and KARP-1 in the telomeric direction. KARP-1 primer extension products were mapped to this CpG island in the correct transcriptional orientation confirming that KARP-1 transcription initiates from this site. Moreover, a p53 response element within the first intron of the KARP-1 transcriptional unit was identified using chromatin immunoprecipitation and antibodies specific to activated forms of p53. These data expand our understanding of this important DNA repair locus. PMID- 11937625 TI - Nuclear actin is associated with a specific subset of hnRNP A/B-type proteins. AB - Pre-mRNP complexes were isolated from rat liver nuclei as 40S hnRNP particles, and actin-binding proteins were collected by DNase I affinity chromatography. The bound proteins were analyzed by 2D gel electrophoresis, and the following five hnRNP A/B-type proteins were identified by tandem mass spectrometry: DBP40/CBF-A (CArG binding factor A), a minor hnRNP A2 variant and three minor hnRNP A3 (mBx) variants. DBP40 was chosen for further analysis of the association of actin with the pre-mRNP complex. It was shown in vitro that purified actin binds to recombinant DBP40 suggesting that the interaction between actin and DBP40 is direct in the pre-mRNP particles. The association of actin with DBP40 was further explored in vivo. It was shown in a transfection study that DBP40 appears both in the nucleus and cytoplasm. Microinjection experiments revealed that DBP40 is exported from the nucleus to the cytoplasm. Finally, RNA-protein and protein protein cross-linking experiments showed that DBP40 interacts with poly(A)(+) RNA as well as actin, both in the nucleus and cytoplasm. We propose that actin associated with DBP40, and perhaps with additional hnRNP A/B-type proteins, is transferred from nucleus to cytoplasm bound to mRNA. PMID- 11937626 TI - A general approach for the use of oligonucleotide effectors to regulate the catalysis of RNA-cleaving ribozymes and DNAzymes. AB - A general approach is described for controlling the RNA-cleaving activity of nucleic acid enzymes (ribozymes and DNAzymes) via the use of oligonucleotide effectors (regulators). In contrast to the previously developed approaches of allosteric and facilitator-mediated regulation of such enzymes, this approach, called 'expansive' regulation, requires that the regulator bind simultaneously to both enzyme and substrate to form a branched three-way complex. Such three-way enzyme-substrate-regulator complexes are catalytically competent relative to the structurally unstable enzyme-substrate complexes. Using the 8-17 and bipartite DNAzymes and the hammerhead ribozyme as model systems, 20- to 30-fold rate enhancements were achieved in the presence of regulators of engineered variants of the above three enzymes, even under unoptimized conditions. Broadly, using this approach ribozyme and DNAzyme variants that are amenable to regulation by oligonucleotide effectors can be designed even in the absence of any knowledge of the folded structure of the relevant ribozyme or DNAzyme. Expansive regulation therefore represents a new and potentially useful technology for both the regulation of nucleic acid enzymes and the detection of specific RNA transcripts. PMID- 11937627 TI - Correlation between chromatin association and transcriptional regulation for the Act3p/Arp4 nuclear actin-related protein of Saccharomyces cerevisiae. AB - Actin-related proteins (Arps), which share a basal structure with actin but have distinct functions, have been found in a wide variety of organisms. While their functions are not yet clear, some Arps are localized in the nucleus and are suggested to contribute to the regulation of transcription. An essential gene of Saccharomyces cerevisiae, Act3p/Arp4, encodes the first identified nuclear Arp, which has been shown to bind to core histones in vitro. Here we have analyzed the in vivo function of Act3p/Arp4 on the his4-912delta promoter. Chromatin immunoprecipitation assays show that Act3p/Arp4 is bound to the entire his4 912delta promoter region. Conditional act3/arp4 mutations affect transcription from the his4-912delta promoter, where decreased Act3p/Arp4 binding and a change in nuclease sensitivity of chromatin were observed, showing the involvement of Act3p/Arp4 in the regulation of gene expression through the organization of chromatin structure. Taken together with the presence of Act3p/Arp4 in chromatin remodeling and histone acetyltransferase complexes, it is suggested that Act3p/Arp4 functions in transcriptional regulation to recruit chromatin remodeling and histone acetyltransferase complexes onto chromatin. PMID- 11937628 TI - Genes on human chromosome 19 show extreme divergence from the mouse orthologs and a high GC content. AB - Mutational rates are known to be variable along the mammalian genome but the extent of this non-random fluctuation and their causes are less well understood. Using 5509 human and mouse orthologous genes with known chromosome positions, it is shown here that there are extreme differences in synonymous evolutionary rates between different human chromosomes when distances are measured using maximum likelihood techniques. In particular, the average synonymous rate of genes located in human chromosome 19 is extremely high (K(s) = 1.243 substitutions/site) compared with the average of all genes (K(s) = 0.729), and significantly different from all other human chromosomes. When genes are sorted according to mouse chromosomes no such large differences are found. Strikingly, almost all genes of human chromosome 19 have very high GC content in humans but not in the mouse orthologs. More generally, correlation analysis shows that genes with very high GC content in humans have experienced the highest synonymous divergencies from the mouse. It is likely that, in such genes, the known relaxation of the isochore structure in rodents has caused an increased accumulation of synonymous substitutions in the mouse lineage, whereas the regions with the highest GC content in the human genome are accordingly maintained by a strong selective pressure. PMID- 11937629 TI - Positional effects of short interfering RNAs targeting the human coagulation trigger Tissue Factor. AB - Chemically synthesised 21-23 bp double-stranded short interfering RNAs (siRNA) can induce sequence-specific post-transcriptional gene silencing, in a process termed RNA interference (RNAi). In the present study, several siRNAs synthesised against different sites on the same target mRNA (human Tissue Factor) demonstrated striking differences in silencing efficiency. Only a few of the siRNAs resulted in a significant reduction in expression, suggesting that accessible siRNA target sites may be rare in some human mRNAs. Blocking of the 3' OH with FITC did not reduce the effect on target mRNA. Mutations in the siRNAs relative to target mRNA sequence gradually reduced, but did not abolish mRNA depletion. Inactive siRNAs competed reversibly with active siRNAs in a sequence independent manner. Several lines of evidence suggest the existence of a near equilibrium kinetic balance between mRNA production and siRNA-mediated mRNA depletion. The silencing effect was transient, with the level of mRNA recovering fully within 4-5 days, suggesting absence of a propagative system for RNAi in humans. Finally, we observed 3' mRNA cleavage fragments resulting from the action of the most effective siRNAs. The depletion rate-dependent appearance of these fragments argues for the existence of a two-step mRNA degradation mechanism. PMID- 11937630 TI - Oct-2 regulates CD36 gene expression via a consensus octamer, which excludes the co-activator OBF-1. AB - The POU domain transcription factor, Oct-2, is essential for the B cell-specific expression of CD36 in mouse B cells. In order to determine how Oct-2 mediates expression of CD36 in B cells, we cloned and analysed the mouse CD36 promoter. In contrast to the human CD36 promoter, the mouse promoter contains a consensus octamer element of the type ATGCTAAT. This octamer element can be bound by either Oct-1 or Oct-2 but requires the expression of Oct-2 to activate transcription in B cells. Mutation of the octamer element renders the CD36 promoter refractory to activation by Oct-2. Furthermore, we demonstrate that the CD36 octamer element does not support recruitment of the B cell-specific co-activator OBF-1 and that CD36 expression is unaffected in primary B cells derived from obf-1(-/-) mice. We conclude that Oct-2 activates CD36 gene expression in mouse B cells via the octamer element in the promoter. Our data also demonstrate that CD36 is the first example of an Oct-2-dependent gene whose expression in B cells is independent of OBF-1. These findings support the notion that Oct-2 regulates gene transcription by both OBF-1-dependent and -independent mechanisms. PMID- 11937631 TI - DNA sequence recognition by the indolocarbazole antitumor antibiotic AT2433-B1 and its diastereoisomer. AB - The antibiotic AT2433-B1 belongs to a therapeutically important class of antitumor agents. This natural product contains an indolocarbazole aglycone connected to a unique disaccharide consisting of a methoxyglucose and an amino sugar subunit, 2,4-dideoxy-4-methylamino-L-xylose. The configuration of the amino sugar distinguishes AT2433-B1 from its diastereoisomer iso-AT2433-B1. Here we have investigated the interaction of these two disaccharide indolocarbazole derivatives with different DNA sequences by means of DNase I footprinting and surface plasmon resonance (SPR). Accurate binding measurements performed at 4 and 25 degrees C using the BIAcore SPR method revealed that AT2433-B1 binds considerably more tightly to a hairpin oligomer containing a [CG](4) block than to an oligomer with a central [AT](4) tract. The kinetic analysis shows that the antibiotic dissociates much more slowly from the GC sequence compared to the AT one. Preferential binding of AT2433-B1 to GC-rich sequences in DNA was independently confirmed by DNase I footprinting experiments performed with a 117 bp DNA restriction fragment. The specific binding sequence 5'-AACGCCAG identified from the footprints was then converted into a biotin-labeled DNA hairpin duplex and compound interactions with this specific sequence were characterized by high resolution BIAcore SPR experiments. Such a combined approach provided a detailed understanding of the molecular basis of DNA recognition. The discovery that the glycosyl antibiotic AT2433-B1 preferentially recognizes defined sequences offers novel opportunities for the future design of sequence-specific DNA-reading small molecules. PMID- 11937632 TI - Calculation of hydrodynamic properties of small nucleic acids from their atomic structure. AB - Hydrodynamic properties (translational diffusion, sedimentation coefficients and correlation times) of short B-DNA oligonucleotides are calculated from the atomic level structure using a bead modeling procedure in which each non-hydrogen atom is represented by a bead. Using available experimental data of hydrodynamic properties for several oligonucleotides, the best fit for the hydrodynamic radius of the atoms is found to be approximately 2.8 A. Using this value, the predictions for the properties corresponding to translational motion and end-over end rotation are accurate to within a few percent error. Analysis of NMR correlation times requires accounting for the internal flexibility of the double helix, and allows an estimation of approximately 0.85 for the Lipari-Szabo generalized order parameter. Also, the degree of hydration can be determined from hydrodynamics, with a result of approximately 0.3 g (water)/g (DNA). These numerical results are quite similar to those found for globular proteins. If the hydrodynamic model for the short DNA is simply a cylindrical rod, the predictions for overall translation and rotation are slightly worse, but the NMR correlation times and the degree of hydration, which depend more on the cross-sectional structure, are more severely affected. PMID- 11937633 TI - Binding of USF to a non-canonical E-box following stress results in a cell specific derepression of the lama3 gene. AB - Expression of the lama3 gene, encoding the laminin alpha3A chain, is restricted to specialized epithelia. We previously showed that lama3 gene expression is controlled by an epithelial enhancer through the cooperative effect of AP-1 binding sites. In fibroblasts, there is no lama3 expression because of the recruitment of a repressor complex absent or inactive in epithelial cells. In this paper, we show evidence that this repression of the lama3 gene is relieved by exogenous and UV-induced USF-1 through its interaction with a non-canonical E box site. Using a chromatin immunoprecipitation assay, we find that UV stress induces USF to bind to the lama3 promoter in vivo. We further demonstrate that this loss of cell specificity is directly related to the accessibility of the E box, resulting in a strong induction in fibroblasts, while expression remains constitutively high in keratinocytes. This accessibility appears to be dependent upon the recruitment of a fibroblastic repressor complex. Therefore, we speculate that anchorage of this repressor complex in fibroblasts modifies the enhancer geometry, allowing USF to interact under stress-inducing conditions with its heptameric binding site. PMID- 11937634 TI - Photoreactivation of UV-induced cyclobutane pyrimidine dimers in the MFA2 gene of Saccharomyces cerevisiae. AB - Photoreactive repair (PR) of cyclobutane pyrimidine dimers (CPDs) was mapped at nucleotide resolution in nucleotide excision repair (NER) proficient and deficient strains for the transcriptionally active and inactive MFA2 gene. Enhanced PR in the control region occurred in areas where no nucleosomes were present, particularly linker regions in the alpha mating type. The presence of excision plus transcriptional activation alleviated this preference, causing repair in the regions that were linker and core in the alpha mating type to be the same in this strain. Transcription had no effect on photoreactive repair in transcribed and downstream regions of MFA2, where similar rates were observed for specific CPDs in both strands. The presence of positioned nucleosomes in alpha mating types revealed slow repair in the nucleosome core, with faster repair occurring at the 3' and 5' edges. These data support the evidence that CPDs are repaired quicker in nucleosome-free regions and at edges of nucleosomes. CPDs in the linker regions are repaired more efficiently in the transcriptionally inactive strains, suggesting that nucleosome movement associated with transcription of MFA2 hampers PR irrespective of the strand. Proficient NER influenced PR in the TATA and Mcm1 binding sites by enhancing it, particularly when transcription was activated. PMID- 11937635 TI - Synergy between cationic lipid and co-lipid determines the macroscopic structure and transfection activity of lipoplexes. AB - The large number of cytofectin and co-lipid combinations currently used for lipoplex-mediated gene delivery reflects the fact that the optimal cytofectin/co lipid combination varies with the application. The effects of structural changes in both cytofectin and co-lipid were systematically examined to identify structure-activity relationships. Specifically, alkyl chain length, degree of unsaturation and the head group to which the alkyl side chain was attached were examined to determine their effect on lipoplex structure and biological activity. The macroscopic lipoplex structure was assessed using a dye-binding assay and the biological activity was examined using in vitro transfection in three diverse cell lines. Lipoplexes were formulated in three different vehicles currently in use for in vivo delivery of naked plasmid DNA (pDNA) and lipoplex formulations. The changes in dye accessibility were consistent with structural changes in the lipoplex, which correlated with alterations in the formulation. In contrast, transfection activity of different lipoplexes was cell type and vehicle dependent and did not correlate with dye accessibility. Overall, the results show a correlation between transfection and enhanced membrane fluidity in both the lipoplex and cellular membranes. PMID- 11937636 TI - A quantitative model of human DNA base excision repair. I. Mechanistic insights. AB - Base excision repair (BER) is a multistep process involving the sequential activity of several proteins that cope with spontaneous and environmentally induced mutagenic and cytotoxic DNA damage. Quantitative kinetic data on single proteins of BER have been used here to develop a mathematical model of the BER pathway. This model was then employed to evaluate mechanistic issues and to determine the sensitivity of pathway throughput to altered enzyme kinetics. Notably, the model predicts considerably less pathway throughput than observed in experimental in vitro assays. This finding, in combination with the effects of pathway cooperativity on model throughput, supports the hypothesis of cooperation during abasic site repair and between the apurinic/apyrimidinic (AP) endonuclease, Ape1, and the 8-oxoguanine DNA glycosylase, Ogg1. The quantitative model also predicts that for 8-oxoguanine and hydrolytic AP site damage, short patch Polbeta-mediated BER dominates, with minimal switching to the long-patch subpathway. Sensitivity analysis of the model indicates that the Polbeta catalyzed reactions have the most control over pathway throughput, although other BER reactions contribute to pathway efficiency as well. The studies within represent a first step in a developing effort to create a predictive model for BER cellular capacity. PMID- 11937637 TI - Species-specific repetitive extragenic palindromic (REP) sequences in Pseudomonas putida. AB - Pseudomonas putida KT2440 is a soil bacterium that effectively colonises the roots of many plants and degrades a variety of toxic aromatic compounds. Its genome has recently been sequenced. We describe that a 35 bp sequence with the structure of an imperfect palindrome, originally found repeated three times downstream of the rpoH gene terminator, is detected more than 800 times in the chromosome of this strain. The structure of this DNA segment is analogous to that of the so-called enterobacteriaceae repetitive extragenic palindromic (REP) sequences, although its sequence is different. Computer-assisted analysis of the presence and distribution of this repeated sequence in the P.putida chromosome revealed that in at least 80% of the cases the sequence is extragenic, and in 82% of the cases the distance of this extragenic element to the end of one of the neighbouring genes was <100 bp. This 35 bp element can be found either as a single element, as pairs of elements, or sometimes forming clusters of up to five elements in which they alternate orientation. PCR scanning of chromosomes from different isolates of Pseudomonas sp. strains using oligonucleotides complementary to the most conserved region of this sequence shows that it is only present in isolates of the species P.putida. For this reason we suggest that the P.putida 35 bp element is a distinctive REP sequence in P.putida. This is the first time that REP sequences have been described and characterised in a group of non-enterobacteriaceae. PMID- 11937638 TI - Recognition of T*G mismatched base pairs in DNA by stacked imidazole-containing polyamides: surface plasmon resonance and circular dichroism studies. AB - An imidazole-containing polyamide trimer, f-ImImIm, where f is a formamido group, was recently found using NMR methods to recognize T*G mismatched base pairs. In order to characterize in detail the T*G recognition affinity and specificity of imidazole-containing polyamides, f-ImIm, f-ImImIm and f-PyImIm were synthesized. The kinetics and thermodynamics for the polyamides binding to Watson-Crick and mismatched (containing one or two T*G, A*G or G*G mismatched base pairs) hairpin oligonucleotides were determined by surface plasmon resonance and circular dichroism (CD) methods. f-ImImIm binds significantly more strongly to the T*G mismatch-containing oligonucleotides than to the sequences with other mismatched or with Watson-Crick base pairs. Compared with the Watson-Crick CCGG sequence, f ImImIm associates more slowly with DNAs containing T*G mismatches in place of one or two C*G base pairs and, more importantly, the dissociation rate from the T*G oligonucleotides is very slow (small k(d)). These results clearly demonstrate the binding selectivity and enhanced affinity of side-by-side imidazole/imidazole pairings for T*G mismatches and show that the affinity and specificity increase arise from much lower k(d) values with the T*G mismatched duplexes. CD titration studies of f-ImImIm complexes with T*G mismatched sequences produce strong induced bands at approximately 330 nm with clear isodichroic points, in support of a single minor groove complex. CD DNA bands suggest that the complexes remain in the B conformation. PMID- 11937639 TI - Downstream sequence elements with different affinities for the hnRNP H/H' protein influence the processing efficiency of mammalian polyadenylation signals. AB - Auxiliary factors likely play an important role in determining the polyadenylation efficiency of mammalian pre-mRNAs. We previously identified an auxiliary factor, hnRNP H/H', which stimulates 3'-end processing through an interaction with sequences downstream of the core elements of the SV40 late polyadenylation signal. Using in vitro reconstitution assays we have demonstrated that hnRNP H/H' can stimulate processing of two additional model polyadenylation signals by binding at similar relative downstream locations but with significantly different affinities. A short tract of G residues was determined to be a common property of all three hnRNP H/H' binding sites. A survey of mammalian polyadenylation signals identified potential G-rich hnRNP H/H' binding sites at similar downstream locations in approximately 34% of these signals. All of the novel G-rich elements tested were found to bind hnRNP H/H' protein and the processing of selected signals identified in the survey was stimulated by the protein both in vivo and in vitro. Downstream G-rich tracts, therefore, are a common auxiliary element in mammalian polyadenylation signals. Sequences capable of binding hnRNP H protein with varying affinities may play a role in determining the processing efficiency of a significant number of mammalian polyadenylation signals. PMID- 11937640 TI - Probabilistic prediction of Saccharomyces cerevisiae mRNA 3'-processing sites. AB - We present a tool for the prediction of mRNA 3'-processing (cleavage and polyadenylation) sites in the yeast Saccharomyces cerevisiae, based on a discrete state-space model or hidden Markov model. Comparison of predicted sites with experimentally verified 3'-processing sites indicates good agreement. All predicted or known yeast genes were analyzed to find probable 3'-processing sites. Known alternative 3'-processing sites, both within the 3'-untranslated region and within the protein coding sequence were successfully identified, leading to the possibility of prediction of previously unknown alternative sites. The lack of an apparent 3'-processing site calls into question the validity of some predicted genes. This is specifically investigated for predicted genes with overlapping coding sequences. PMID- 11937642 TI - Investigation of the multiple anchors approach in oligonucleotide microarray preparation using linear and stem-loop structured probes. AB - Enzyme-mediated reactions are a useful tool in mutation detection when using a microarray format. Discriminating probes attached to the surface of a DNA chip have to be accessible to target DNA and to the enzyme (ligase or polymerase) that catalyses the formation of a new phosphodiester bond. This requires an appropriate chemical platform. Recently, an oligonucleotide hairpin architecture incorporating multiple phosphorothioate moieties along the loop has been proposed as an effective approach to solid-phase minisequencing. We have explored in depth several variables (stem length, number of phosphorothioates, stem-loop architecture versus linear structure) involved in this strategy by using a solid phase ligation reaction. Microarrays were fabricated either from aminosilyl modified glass or from aminated polymeric surfaces made of poly-lysine. Both platforms were bromoacetylated and reacted with thiophosphorylated oligonucleotides. The resulting microarrays were tested using either a synthetic template or a PCR-amplified 16S rRNA genomic region as the target sequence. Our results confirm the robustness of the proposed chemistry. We extend its range of application to solid-phase ligation, demonstrating the effectiveness of multiple anchors and suggest that linear oligonucleotides incorporating multiple phosphorothioates are equivalent to their hairpin-structured counterparts. PMID- 11937641 TI - Gene expression changes in response to E2F1 activation. AB - The p16/RB/E2F regulatory pathway, which controls transit through the G1 restriction point of the cell cycle, is one of the most frequent targets of genetic alterations in human cancer. Any of these alterations results in the deregulated expression of the transcription factor E2F, one of the key mediators of cell cycle progression. Under these conditions, E2F1 also participates in the induction of apoptosis by a p53-dependent pathway, and independently of p53. Recently, we identified the p53-homolog p73 as a first direct target of p53 independent apoptosis. Here, we used a cDNA microarray to screen an inducible E2F1-expressing Saos-2 cell line for E2F1 target genes. Expression analysis by cDNA microarray and RT-PCR revealed novel E2F1 target genes involved in E2F1 regulated cellular functions such as cell cycle control, DNA replication and apoptosis. In addition, the identification of novel E2F1 target genes participating in the processes of angiogenesis, invasion and metastasis supports the view that E2F1 plays a central role in many aspects of cancer development. These results provide new insight into the role of E2F1 in tumorigenesis as a basis for the development of novel anti-cancer therapeutics. PMID- 11937645 TI - Case reports, case series and systematic reviews. PMID- 11937643 TI - Generating segmental mutations in haloalkane dehalogenase: a novel part in the directed evolution toolbox. AB - Directed evolution techniques allow us to genuinely mimic molecular evolution in vitro. To enhance this imitation of natural evolutionary processes on a laboratory scale in even more detail, we developed an in vitro method for the generation of random deletions and repeats. The pairwise fusion of two fragments of the same gene that are truncated by exonuclease BAL-31 either at the 3' or 5' side results in a deletion or a repeat at the fusion point. Although in principle the method randomly covers the whole gene, it can also be limited to a predefined area in the sequence by controlling the level of the initial truncation. To test the procedure and to illustrate its potential, we used haloalkane dehalogenase from Xanthobacter autotrophicus GJ10 (DhlA) as a model enzyme, since the adaptation of this enzyme towards new substrates is known to occur via the generation of this type of mutation. The results show that the mutagenesis method presented here is an effective tool for accessing formerly unexplorable sequence space and can contribute to the success of future directed evolution experiments. PMID- 11937646 TI - Peripheral vascular disease and Virchow's triad for thrombogenesis. PMID- 11937647 TI - Unicentric Castleman's disease complicated by systemic AA amyloidosis: a curable disease. AB - BACKGROUND: Castleman's disease (angiofollicular lymph node hyperplasia) is a group of rare lymphoproliferative disorders sharing characteristic clinical and histological features, and usually accompanied by a marked systemic inflammatory response. All types may be complicated by acquired systemic amyloidosis, usually of AA type, but occasionally of AL type associated with monoclonal gammopathy. DESIGN: Descriptive study of five patients with unicentric Castleman's disease complicated by systemic AA amyloidosis. METHODS: A diagnosis of amyloidosis was confirmed by microscopy and immunohistochemical staining. Serum concentrations of C-reactive protein (CRP) and serum amyloid A protein (SAA) were measured by immunoassays. Radiolabelled serum amyloid P component scintigraphy was used to monitor the progress of amyloid deposition. RESULTS: In four patients the primary diagnosis was made only after years of investigation of systemic symptoms. The tumours were resected in all cases, leading to remission of the systemic inflammatory state. Long-term follow-up in four patients, including scintigraphy, showed regression of amyloid deposits. DISCUSSION: This rare but usually fatal condition can be cured surgically even in advanced cases. Awareness of the diagnosis and its correct management are important in investigation of patients with unexplained systemic symptoms, especially associated with systemic amyloidosis. PMID- 11937648 TI - Anticonvulsant drug use and low bone mass in adults with neurodevelopmental disorders. AB - BACKGROUND: Use of anticonvulsant drugs among adults with neurodevelopmental disorders may be an important risk factor for both osteoporosis and skeletal fractures. AIM: To determine the relationship between anticonvulsant drug use and both low bone mass and bone fractures in such adults. DESIGN: Cross-sectional study of 273 adults with neurodevelopmental disorders, 40% of whom were receiving one or more anticonvulsant drugs. SETTING: Single Canadian long-term care facility. METHODS: Demographic data were abstracted from each resident's chart in a standardized manner, including body mass, degree of mobility, major falls within the previous 12 months, and all medications. Quantitative calcaneal ultrasonography was performed on each resident without knowledge of their current drug use. The Quantitative Ultrasound Index was employed to express 'bone stiffness'. Low bone mass was defined as a T-score 2.5 SDs below the norm for young healthy adults. RESULTS: Compared to non-users (15.5%), low bone mass was more prevalent among those taking either one (20.3%; OR 1.7, 95%CI 0.6-4.4) or two or more anticonvulsant agents (42.2%, OR 5.9, 95%CI 2.2-16.2). The risk of recent skeletal fractures was not significantly greater in those taking a single anticonvulsant than in non-users (28.6% vs. 21.6%; OR 1.0, 95%CI 0.4-2.8), but tended to be higher in those taking two or more (48.7%; OR 2.2, 95%CI 0.8-5.9). CONCLUSIONS: Adults with neurodevelopmental disorders residing in a long-term care facility have a high rate of both low bone mass and skeletal fractures, especially with concomitant use of anticonvulsant drugs. These individuals should be assessed for the presence of low bone mass, and may warrant prophylactic treatment against bone loss, including calcium and vitamin D supplementation. PMID- 11937649 TI - A comparison of lorazepam and diazepam as initial therapy in convulsive status epilepticus. AB - BACKGROUND: Previous trials have suggested lorazepam may be superior to diazepam as first-line treatment of convulsive status epilepticus (CSE), with improved seizure outcome, and a lower incidence of side-effects. Many published guidelines however still recommend diazepam. AIM: To compare the efficacy, safety and cost of lorazepam compared to diazepam, in adults with CSE. DESIGN: Retrospective case note audit. METHODS: Cases of CSE were retrospectively identified over two 18 month periods either side of the introduction of a new management protocol in May 1997, in which lorazepam 4 mg i.v. was substituted for diazepam 10 mg i.v. as first-line treatment for CSE. Diagnostic codes for all admissions and casualty presentations of patients over 16 years of age were examined for primary or secondary codes including 'epilepsy', 'fits' or 'status epilepticus'. Medical records and casualty notes were reviewed to identify CSE cases. Treatment groups were compared using ANOVA and a Tukey post hoc analysis. Treatment success was defined as cessation of seizures without recurrence in the subsequent 12 h. RESULTS: In both premonitory and established CSE, both drugs were equally effective at terminating seizures, but significantly fewer seizure recurrences followed lorazepam, and fewer repeat doses were needed. There were no differences in reported adverse events or in drug costs. DISCUSSION: We recommend that lorazepam be the first-line therapy in preference to diazepam in adults with CSE. PMID- 11937652 TI - The nature of truth: Simpson's Paradox and the limits of statistical data. PMID- 11937650 TI - The influence of serum cytokines and growth factors on osteoclast formation in Paget's disease. AB - BACKGROUND: Osteoclasts are multinucleated cells (MNCs) that form from circulating mononuclear precursors in the presence of the receptor activator of nuclear factor kappa B-ligand (RANKL) and macrophage-colony stimulating factor (M CSF). AIM: To determine whether cytokines and growth factors influence RANKL/M CSF induced osteoclastogenesis and bone resorption in Paget's disease. DESIGN: Prospective case-control study. METHODS: Serum levels of M-CSF, interleukin (IL) 1 beta, IL-6 and tumour necrosis factor-alpha (TNFalpha) were measured in 13 Paget's disease patients and 8 normal controls. The effect of serum from Paget's patients on osteoclast formation was also assessed. RESULTS: Serum levels of IL-1 beta, IL-6 and TNFalpha were low or undetectable in Paget's disease patients and normal controls. Levels of M-CSF were significantly increased in Paget's patients who were not currently under treatment. In Paget's patients under treatment, serum M-CSF levels were not significantly different from normal controls. The addition of serum from untreated Paget's patients dose-dependently increased RANKL-induced osteoclast formation and lacunar resorption in normal monocyte cultures; elevated IL-6 levels were found in the supernatant and the addition of a specific antibody to human IL-6 blocked the increase in osteoclast formation and resorption. Serum from untreated Paget's patients also induced osteoclast formation in the absence of exogenous M-CSF; an antibody specific to human M-CSF abolished this effect. DISCUSSION: Both M-CSF and IL-6 play a major role in osteoclast formation and bone resorption in Paget's disease and measurement of serum M-CSF may provide a useful indicator of disease activity. PMID- 11937651 TI - Complications and cardiovascular risk factors in South Asians and Europeans with early-onset type 2 diabetes. AB - BACKGROUND: Type 2 diabetes is a major cardiovascular risk factor, and early onset (<40 years) type 2 diabetes is becoming more common. AIM: To determine the prevalence of complications, and cardiovascular risk factors at diagnosis, in early-onset type 2 diabetes, and to compare these between South Asians and Europeans. DESIGN: Prospective study of newly-diagnosed type 2 diabetes patients aged <40 years, attending hospital and primary care clinics 1999-2001. METHODS: Patients were assessed for signs of macrovascular disease, retinopathy, neuropathy and nephropathy. Cardiovascular risk factors were also determined. RESULTS: Overall, 292 patients were enrolled (165 South Asians). Macrovascular disease was more prevalent in South Asians (15.7% vs. 9.4%, p<0.001), as was microvascular disease (27.3% vs. 16.5%, p<0.001), including retinopathy (17.5% vs. 7.9%, p<0.001), and nephropathy (18.1% vs. 7.8%, p<0.001). South Asians had trends towards greater waist:hip ratio (0.95 vs. 0.90), and higher blood pressure (127/80 vs. 123/76 mmHg). HDL cholesterol was lower (1.0 vs. 1.3 mmol/l, p<0.001) and fasting triglycerides higher (1.9 vs. 1.5 mmol/l, p<0.001) in South Asians. Absolute CHD risk was significantly higher in South Asians (16.9% vs. 13.7%, p<0.001). DISCUSSION: Complications were common at diagnosis, with a quarter of all patients having evidence of at least one diabetic complication. South Asians had a higher prevalence of established macrovascular and microvascular disease, compared to Europeans, and a higher risk of CHD, predominantly because of lower HDL cholesterol and higher blood pressure. PMID- 11937653 TI - A brief history of the clinical thermometer. PMID- 11937654 TI - Recurrent venous thromboses, anti-cardiolipin antibodies and Crohn's disease. PMID- 11937655 TI - Asthma attacks and thunderstorms. PMID- 11937656 TI - Severe osteoporosis and autonomous hyperparathyroidism: making the numbers add up. PMID- 11937657 TI - Variety. PMID- 11937658 TI - What's in a name? PMID- 11937659 TI - Some aspects of anaesthesiology. PMID- 11937660 TI - Epidemiological study of multiple sclerosis in Hong Kong Chinese: questionnaire survey. AB - OBJECTIVE: To study the epidemiology of multiple sclerosis in Hong Kong Chinese. DESIGN: Cross-sectional questionnaire survey. SETTING: Neurology and paediatric neurology departments in Hong Kong from January through June 1999. PARTICIPANTS: All confirmed multiple sclerosis patients. MAIN OUTCOME MEASURES: Demographic data, investigation results, Kurtzke's Expanded Disability Status Scale during the last follow-up visit, number of relapses between 1997 and 1998, and treatments used/currently in use. RESULTS: Fifty-three Chinese multiple sclerosis patients were identified. The prevalence was thus estimated to be 0.77 per 100,000 population. This low prevalence was also noted in other multiple sclerosis studies from South-East Asia (range, 0.8-4 per 100,000 population). The female to male ratio among the Chinese multiple sclerosis sufferers was 9.6:1, a figure somewhat higher than that reported in the other studies from South-East Asia (range, 3.2-6.6:1). The Chinese multiple sclerosis patients in this study also had a high spinal cord involvement (66%) and a low presence of cerebrospinal fluid oligoclonal banding (40%). These findings were different from those in Caucasian multiple sclerosis patients. CONCLUSION: Multiple sclerosis in Hong Kong Chinese has a low prevalence, a high female to male ratio, and a low cerebrospinal fluid oligoclonal banding presence. PMID- 11937661 TI - Review of children with severe trauma or thermal injury requiring intensive care in a Hong Kong hospital: retrospective study. AB - OBJECTIVE: To study the injury pattern of children admitted for management of severe trauma or thermal injury. DESIGN: Retrospective review. SETTING: Paediatric intensive care unit of a regional hospital, Hong Kong. PATIENTS: Twenty-eight children were admitted under this category from July 1996 to December 1999. MAIN OUTCOME MEASURES: Mechanisms, severity, and circumstances of injury. RESULTS: Road traffic accident was the most common cause of admission, followed by thermal injury, accidental fall, and non-accidental injury. However, children with non-accidental injury were admitted in a significantly more severe condition, as measured by the paediatric risk of mortality score, than those admitted for the other three reasons. Non-accidental injury was also associated with significantly higher morbidity and mortality than the other causes of admission. CONCLUSIONS: During the 42-month study period, trauma and thermal injury accounted for 7% of all admissions to the paediatric intensive care unit. Road traffic accident was the most common reason, while non-accidental injury accounted for the most serious injury. Detailed analysis of these cases identified certain preventable risk factors. PMID- 11937662 TI - Patient absconding behaviour in a public general hospital: retrospective study. AB - OBJECTIVE: To identify characteristics of patients who abscond from general hospital wards, and to determine patient outcomes. DESIGN: Retrospective study. SETTING: In-patient wards of a public general hospital, Hong Kong. SUBJECTS AND METHODS: Incident reports of 116 absconding episodes over a 20-month period from 1 November 1998 to 30 June 2000 were reviewed. Clinical characteristics, time and reason for absconding, destination of patient, outcomes, and adverse events were recorded. RESULTS: The majority of patients who absconded were middle-aged males admitted through the Accident and Emergency Department to the specialty of general medicine. Most incidences occurred within 24 hours of admission while in the emergency admission wards. Twelve percent of incidents were repeated episodes of absconding. The most common clinical diagnoses given to this patient group were drug overdose, intoxication, and soft tissue injury. Known drug addicts (29.3%) formed a substantial proportion of the patient group. Forty- seven patients returned to the ward within a few hours, while a further nine (7.8%) re attended the Accident and Emergency Department of the hospital within 4 days. Several adverse outcomes were recorded: one patient died following a fall from height and two patients committed criminal offences. CONCLUSION: Patient absconding incidents are an important issue in hospital risk management. They can delay the delivery of appropriate medical treatment and may lead to other adverse patient outcomes, in addition to potential medicolegal consequences. PMID- 11937663 TI - Group A streptococcal infection in patients presenting with a sore throat at an accident and emergency department: prospective observational study. AB - OBJECTIVES: To determine the prevalence of group A streptococcal infection and to evaluate the predictive value of clinical findings and rapid streptococcal antigen detection testing in patients presenting with a sore throat or suspected clinically to have acute pharyngitis. DESIGN: Prospective observational study. SETTING: Accident and emergency department of a public hospital, Hong Kong. PATIENTS: All patients presenting with a sore throat as the chief complaint, or suspected clinically to have acute pharyngitis, from April to September 2000. MAIN OUTCOME MEASURES: Demographic data, clinical features, microbiological throat culture results, and rapid streptococcal antigen detection (Accustrip) test results. RESULTS: Of 1449 patients recruited during the 6-month study period, only 44 (3.0%) had positive throat cultures for group A beta-haemolytic streptococcus. The majority of group A beta-haemolytic streptococci were isolated from patients between the age of 3 and 60 years. Clinical findings other than an absence of cough were found to be unhelpful in predicting group A beta-haemolytic streptococcal throat infection. The sensitivity of the rapid group A streptococcal antigen detection test was 52.6% and the specificity was 98.2%. CONCLUSIONS: The prevalence of group A beta-haemolytic streptococcus in patients presenting with a sore throat, or suspected clinically of having acute pharyngitis, was low. If empirical antibiotics were given to all such patients, 97% of them would be unnecessarily treated. Age and absence of cough were the only clinical findings helpful in predicting the presence of group A beta haemolytic streptococcal throat infection. The rapid group A streptococcal antigen detection test can provide a quick guide to clinicians on the necessity of antibiotic therapy. However, a confirmatory throat culture backup is recommended for patients with a negative test result. PMID- 11937664 TI - Preparation of patients for anaesthesia - achieving quality care. AB - Implementation of anaesthesia begins with a preoperative assessment of the surgical patient and development of an anaesthetic plan. Preparation of the patient includes the preoperative assessment, review of preoperative tests, optimisation of medical conditions, adequate preoperative fasting, appropriate premedication, and the explanation of anaesthetic risk to patients. The goals of preoperative preparation are to reduce the morbidity of surgery, to increase the quality while decreasing the cost of perioperative care, and to return the patient to desirable functioning as quickly as possible. A knowledgeable anaesthesiologist is the 'final clinical gatekeeper', who coordinates perioperative management and ensures that the patient is in the optimal state for anaesthesia and surgery. PMID- 11937665 TI - Local anaesthesia outside the operating room. AB - An increasing number of minor surgical procedures are performed under local anaesthesia in clinical settings outside the operating room, where monitoring and resuscitation equipment--as well as personnel skilled in resuscitation--may not be readily available. Serious adverse effects and even fatalities may result from the use of local anaesthetic agents, arising from a variety of causes such as systemic toxicity, allergy, vasovagal syncope, and reaction to additives present in the local anaesthetic. This article briefly reviews the pharmacology of local anaesthetic agents, and describes various techniques commonly used for local anaesthesia, with special emphasis on safety. Clinical features of toxicity, and its differential diagnosis and management, are also discussed. PMID- 11937666 TI - Safety and comfort during sedation for diagnostic or therapeutic procedures. AB - Sedation during diagnostic or therapeutic procedures must be safe and comfortable for patients. To achieve this, additional suitably qualified staff must be available throughout the procedure to administer sedation and monitor the patient. Anaesthesiologists possess the necessary knowledge and skills to perform sedation safely but are often unavailable. Non-anaesthesiologists performing sedation should be fully trained in the physiology of sedation, the pharmacology of sedatives and analgesics, the monitoring of patients, and in airway support, ventilatory care, and cardiopulmonary resuscitation. The presence of an anaesthesiologist is desirable when dealing with patients at high-risk of complications. Good sedation practice involves presedation assessment and optimal selection of patients, careful monitoring and support from dedicated staff, and adherence to recovery and discharge criteria. PMID- 11937668 TI - The application of clinical simulation in crisis management training. AB - Since it was first introduced more than 30 years ago, clinical simulation has become a popular tool for medical training, particularly in crisis management. The modern high-fidelity patient simulator consists of a whole-body mannequin with integrated electronic patient monitoring; it is controlled by computers capable of simulating numerous clinical scenarios and patient characteristics, and reacting to various interventions appropriately. Simulator training is theoretically superior to conventional training in management of rare crisis situations, as it allows unlimited practice in a safe yet familiar environment. Training in clinical skills can be developed, together with competency in crisis management behaviours such as leadership and communication skills. Although there is evidence demonstrating the popularity, reliability, and validity of simulator training, its superiority over conventional training has not been proven, and research in this area is required. PMID- 11937667 TI - Herbal medicine and anaesthesia. AB - Herbal medicines are increasingly used in both western and Chinese societies. This is partly attributed to the alleged limitations of scientific medicine in the cure and control of chronic diseases. Many patients do not disclose that they have used herbs before surgery and hence their physicians remain unaware of the potential herb-drug interactions. With respect to anaesthesia, herbs can cause coagulation disorders, cardiovascular side-effects, water and electrolyte disturbances, endocrine effects, hepatotoxicity, and prolongation of the effects of anaesthetic agents. Anaesthesiologists should obtain a history of herbal medicine use from patients and work out the adverse perioperative herb-drug interactions in advance of the actual operation. If in doubt, the herbal medicine should be stopped for 2 weeks prior to anaesthesia and surgery. PMID- 11937669 TI - Respiratory insufficiency in a Chinese adult with mitochondrial myopathy. AB - Mitochondrial myopathy is an important but uncommon cause of respiratory insufficiency in adults. We report the first case of respiratory insufficiency associated with adult-onset mitochondrial myopathy seen in a Chinese adult in Hong Kong. The patient presented with peripheral oedema and shortness of breath over 2 to 3 days. There was a history of gradual progressive limb weakness over approximately 2 years, hypertrophic cardiomyopathy, intermittent diarrhoea, and weight loss. The diagnosis was made by skeletal muscle biopsy and molecular study, which revealed the A3243G point mutation. PMID- 11937670 TI - Pulmonary embolism presenting as disseminated intravascular coagulation. AB - We report an unusual case of disseminated intravascular coagulation. Occult pulmonary embolism is a recognised cause of disseminated intravascular coagulation. Unexplained shock should prompt the physician to search for a thrombotic cause such as pulmonary thromboembolism. PMID- 11937671 TI - Treatment of a ruptured iliac aneurysm with an endoluminal stent graft. AB - The management of a 79-year-old man presenting with a ruptured common iliac aneurysm is described. The patient had multiple medical problems including hypertension, ischaemic heart disease, and atrial fibrillation, as well as a left hemispheric stroke 5 years previously. Traditional open surgery was judged unsuitable in this case because of the predicted poor outcome. The patient was subsequently treated successfully with endoluminal stent grafting. PMID- 11937672 TI - Lytic skull metastasis secondary to thyroid carcinoma in an adolescent. AB - We report an unusual case of skull metastasis secondary to thyroid carcinoma in an adolescent girl. The 18-year-old presented with an occipital scalp swelling of 5 years' duration. She reported having thyroid surgery in mainland China 10 years previously. Radiological investigations on presentation demonstrated a lytic hypervascular skull lesion. Preoperative angiography and embolisation was followed by surgical excision. Pathological examination showed the lesion to be a thyroid carcinoma with a predominantly follicular pattern and a completion hemithyroidectomy was subsequently performed. Computed tomography of the thorax showed small micronodules in both lung fields compatible with metastases. The patient was given whole body iodine-131 internal radiation treatment and subsequently commenced thyroid-stimulating hormone-suppressive treatment with thyroxine. The management of thyroid carcinoma and subsequent skull metastasis in children and adolescents is reviewed and controversial points are highlighted. PMID- 11937673 TI - Interacting with the media. PMID- 11937674 TI - The link between bacterial radiation resistance and cold adaptation. PMID- 11937675 TI - Warring ants: lessons from Lanchester's laws of combat? PMID- 11937676 TI - Vernalization: the flower school. PMID- 11937678 TI - Stem cell identity: life is plastic, it's fantastic! PMID- 11937677 TI - The ascent of nucleotide cyclases: conservation and evolution of a theme. PMID- 11937679 TI - Developmental mechanisms: putting genes in their place. PMID- 11937680 TI - Genes encoding chimeras of Neurospora crassa erg-3 and human TM7SF2 proteins fail to complement Neurospora and yeast sterol C-14 reductase mutants. AB - The human gene TM7SF2 encodes a polypeptide (SR-1) with high sequence similarity to sterol C-14 reductase, a key sterol biosynthetic enzyme in fungi, plants and mammals. In Neurospora and yeast this enzyme is encoded by the erg-3 and erg24 genes respectively. In an effort to demonstrate sterol C-14 reductase activity for SR-1 we constructed six recombinant genes coding for chimeras of the Neurospora erg-3 and SR-1 protein sequences and tested them for complementation of the Neurospora erg-3 mutant. To our surprise, all the chimeras failed to complement erg-3. A few of the chimeric proteins were also tested against the yeast erg24 mutant, but again there was no complementation. We discuss some reasons that might account for these unexpected findings. PMID- 11937681 TI - DNA synthesis in the imaginal wing discs of the American bollworm Helicoverpa armigera (Hubner). AB - The effect of two insect growth regulators of plant origin viz. plumbagin and azadirachtin and the ecdysteroids 20-hydroxyecdysone, makisterone A and a phytoecdysteroid on DNA synthesis in imaginal wing discs of day 4 final instar Helicoverpa armigera larvae was studied. DNA synthesis increased with increase in time of incubation up to 8 h and decreased later without the addition of moulting hormone. Addition of 20-hydroxyecdysone supported long term acquisition of competence for DNA synthesis in the wing discs. Both DNA synthesis and protein content were drastically reduced in plumbagin and azadirachtin-treated insects. Under in vitro conditions, plumbagin had a more pronounced inhibitory effect than azadirachtin. All the ecdysteroids tested, viz. makisterone A, 20-hydroxyecdysone and the ecdysteroidal fraction from the silver fern Cheilanthes farinosa enhanced DNA synthesis. PMID- 11937682 TI - Enthalpy-entropy compensation: a phantom phenomenon. AB - Enthalpy-entropy compensation is the name given to the correlation sometimes observed between the estimates of the enthalpy and entropy of a reaction obtained from temperature-dependence data. Although the mainly artefactual nature of this correlation has been known for many years, the subject enjoys periodical revivals, in part because of the frequent excellence of the correlation. As with other cases of impossibly good correlation between two biological variables, the explanation is that what purports to be two variables are very largely the same variable looked at in two different ways. PMID- 11937683 TI - Epigenetics of dominance for enzyme activity. AB - We have isolated and purified two parental homodimers and a unique heterodimer of acid phosphatase [coded by Acph-1 (1.05)(F) and Acph-1 (0.95)(S)] from isogenic homozygotes and heterozygotes of Drosophila malerkotliana. F and S produce qualitatively different allozymes and the two alleles are expressed equally within and across all three genotypes and F and S play an equal role in the epigenetics of dominance. Subunit interaction in the heterodimer over a wide range of H+ concentrations accounts for the epigenetics of dominance for enzyme activity. PMID- 11937684 TI - Role of HLA antigens in Rh (D) alloimmunized pregnant women from Mumbai, Maharashtra, India. AB - Immunogenetic studies in various diseases provide potential genetic markers. We have studied the incidence of HLA A, B, C, DR and DQ loci antigen in Rh (D) antigen isoimmunized mothers compared to those nonimmunized isoimmunized Rh negative mothers. Seventy six mothers who were immunized to Rh (D) antigen due to pregnancy (responders) and fifty four mothers who did not develop Rh (D) isoimmunization despite positive pregnancies (nonresponders) were selected for the study. Standard methods of serological HLA typing, ABO and Rh (D) groups, and screening for Rh D antibodies were used. 392 unrelated individuals from the population were compared as controls. In addition 45 unrelated individuals from the same population were typed for HLA DRB and DQB gene using PCR-SSP kits. The genotype frequencies of HLA A2, A3, A28, B13, B17, B35, B52, B60, Cw2, Cw6, DR4, and DQ3 were significantly increased, while the frequencies of the HLA A11, A29, A31, B7, B37, B51, Cw1 and DR9 were decreased in the responder women when compared to the non-responder women. HLA A30 (19) split antigen was not identified in immunized women while HLA A23 (9) split antigen was not identified in non immunized women. HLA A3, B17, Cw2 and DR4 showed a significant relative risk among the immunized responder women. When compared with Rh immunized women (responders) reported from USA, England and Hungary the phenotype frequencies of HLA A11, A24, A28, B5, B17, B40, DR2 and DR5 were increased while HLA A23, B8, B18, and DR6 were decreased in the Indian Rh immunized women. Two locus haplotype frequency analysis observed among the responders women revealed that among the significant haplotypes expressed A2-B5, B7-Cw1, DR2-DQ1 were highly significant haplotypes in positive linkage, while A1-B5, and A1-B7 were in significant negative linkage disequilibrium. The haplotype frequencies were 46 micromol/l (8.2 microg/ml; p < 0.001). In contrast, lower theophylline concentrations (about 28 micromol/l or 5 microg/ml), either in vitro or in vivo, caused significant increases in neutrophil activities. Theophylline concentrations in the high therapeutic range (84 micromol/l or 15 microg/ml) cause dose-dependent reductions in neutrophil chemiluminescence, chemotaxis, and random motility. Cells from preterm infants are particularly sensitive to this effect. In contrast, theophylline concentrations in the low therapeutic range (28 micromol/l or 5 microg/ml) cause increased neutrophil activities. Altered neutrophil activity in newborns related to theophylline treatment may affect the infants' response to infection as well as the incidence of inflammatory diseases. PMID- 11937724 TI - Prophylactic administration of porcine-derived lung surfactant is a significant factor in reducing the odds for peri-intraventricular haemorrhage in premature infants. AB - We hypothesized that prophylactic treatment with a porcine-modified lung surfactant (PLS) reduces the rate of peri-intraventricular haemorrhage (PIVH) more than rescue treatment. We performed a meta-analysis of three prophylactic versus rescue trials conducted with PLS in four countries using individual data. Overall (grades 1-4) or severe (grades 3 and 4) PIVH of 671 newborns was the outcome. A logistic regression analysis was performed. Prophylactic exposure to PLS was a significant independent factor in reducing the incidence of overall (OR 0.65; 95% CI 0.47-0.90) and severe (OR 0.56; 95% CI 0.35-0.89) PIVH. Moreover, for severe PIVH, the adjusted OR for outborn babies exposed to prophylactic treatment with PLS was highly significant (OR 0.11; 95% CI 0.02-to 0.49). The results we obtained show that prophylactic treatment with PLS reduces the rate of PIVH more than rescue treatment. PMID- 11937725 TI - Sows' milk attenuates dexamethasone-induced reductions in liver docosahexaenoic acid. AB - The objective was to determine the effects of dexamethasone (Dex) on growth and arachidonic (AA) and docosahexaenoic (DHA) acid status. Piglets were randomized to suckling or formula feeding plus Dex or placebo treatment for 15 days. Weight and length at the beginning and end of study was used to assess growth. After 15 days, AA and DHA in phosphatidylethanolamine (PE) and phosphatidylcholine (PC) were measured in liver and plasma and total AA and DHA in forebrain and brainstem. Dex treatment significantly reduced weight and length. Dex reduced liver PC and PE DHA in the formula group, but the reduction in the suckled group was not significant. PC AA and DHA were positively related in liver and plasma. A main effect of Dex to reduce AA and DHA was observed in forebrain but not in brainstem. Sows' milk attenuated the Dex-induced reduction in liver DHA but not forebrain AA and DHA or somatic growth. PMID- 11937726 TI - Endotoxemia and the effects of dopamine on renal functions of neonatal piglets. AB - In this study, we observed the effects of moderate and high doses of dopamine on the renal functions of neonatal piglets during endotoxic shock. We found that fluid therapy alone was better at maintaining cardiac index and preventing elevation of systemic vascular resistance, than dopamine at 10 and at 20 microg/kg/min. Furthermore, urine output and glomerular filtration rate were reduced by dopamine. Following endotoxin administration dopamine decreased SVR and maintained a CI better than fluid alone. However, in spite of a better CI, greater deterioration in renal functions occurred in the dopamine groups as compared to the fluid group. PMID- 11937727 TI - Neuroendocrine and immune responses to acute endotoxemia in suckling and weaned piglets. AB - The objective of this study was to characterize effects of weaning stress on behavioural, endocrine and immune responses to acute peripheral lipopolysaccharide (LPS) challenge in neonatal pigs. Weaning in 28-day-old piglets was accompanied by a significant increase in ACTH concentrations (p = 0.0378) and an increase in basal cortisol level (p = 0.0135). There was also a significant suppressive effect on lymphocyte proliferation in response to concanavalin A (p = 0.0048) in newly weaned piglets. Peripheral administration of LPS induced vomiting, diarrhoea and somnolence in both suckling and weaned piglets. The frequency of these signs of sickness was significantly higher in weaned piglets compared with suckling piglets (p = 0.0049). Additionally, LPS significantly increased plasma concentrations of TNF-alpha, cortisol and ACTH. While weaned piglets reacted to LPS with a higher release of ACTH (p = 0.0239) and cortisol (p = 0.0015) than suckling piglets there was no significant effect of weaning on the magnitude of TNF-alpha. The present data indicate that weaning suppresses the lymphocyte function, causes changes in endocrine regulation and has a substantial effect on the behavioural and endocrine response to an acute peripheral LPS challenge; consequently it could increase disease susceptibility. PMID- 11937728 TI - Neonatal intestinal occlusion due to duodenal duplication in association with malformed gallbladder sludge. AB - Duodenal duplications are rare observations which can be diagnosed during early pregnancy via US scan. In the neonate they are often cause for intestinal occlusions. Surgical treatment can be limited by the duplication's anatomical interrelationships with adjacent organs. Biliary sludge is an uncommon finding in the first year of life, and can readily regress spontaneously. The association between duodenal duplication and sludge has never been described in the literature in the neonatal period. Here, for the first time, we report on the case of a newborn infant with cystic duplication of the duodenum associated with sludge in a misshapen gallbladder. PMID- 11937729 TI - Effect of low-dose oral erythromycin on gastric aspirates in ventilated neonates less than 32 weeks of gestation. Preliminary results. AB - In this prospective study, the 24-hour gastric aspirate volume was carefully recorded before, 24 and 48 h after administering 1.7 mg/kg/8-hourly of oral erythromycin to 16 ventilated neonates less than 32 weeks of gestation. Their median gestational age was 28.5 weeks (range 23-31 weeks), their median birthweight was 1,045 g (range 690-1,560 g) and the median day of life at which erythromycin was commenced was 9.5 days (range 4-16 days). Prior to administering erythromycin median 24-hour gastric aspirate volume, expressed as a percentage of the milk volume given over the same period, was 38.5% (range 20.0-100%). It was significantly lower 24 h (median 12%, range 0-41%, p = 0.0004) and 48 h (median 5%, range 0-21%, p = 0.0004) after commencing erythromycin. There was also significant reduction of gastric aspirate volume between 24 and 48 h after commencing erythromycin (p = 0.0024). Milk volume increment over the same period was not significant (p = 0.1022). These preliminary results warrant further evaluation through a randomised controlled trial. PMID- 11937730 TI - What Is notalgia paresthetica? PMID- 11937731 TI - Immunohistochemical findings in notalgia paresthetica. AB - BACKGROUND: Notalgia paresthetica (NP) is a sensory neuropathy the pathogenesis of which is not yet completely elucidated. OBJECTIVE: The aim of this study was to investigate the histopathological changes in NP with special emphasis on cutaneous innervation. METHODS: Along with site-matched biopsies from 5 healthy individuals, lesional skin biopsies from 14 cases of NP and biopsies from contralateral nonlesional skin in 9 of these cases were stained with hematoxylin eosin and Congo red. For immunohistochemical analysis, all samples were stained with two general neural markers (S-100 protein and protein gene product 9.5) and two neuropeptides (vasoactive intestinal polypeptide and substance P). RESULTS: Light microscopy was compatible with postinflammatory hyperpigmentation. Immunohistochemistry did not reveal a significant difference in the staining pattern of lesional skin and control tissue (p > 0.05). Although not reaching statistical significance, the percentage of cases which showed no staining was higher in the group of patients with more chronic NP. CONCLUSION: The finding of less immunohistochemical staining in cases with more chronicity could be of clinical importance and is worth investigating further. PMID- 11937732 TI - Psoriatic lesional keratinocytes promote the maturation of human monocyte-derived Langerhans cells. AB - BACKGROUND: Langerhans cells (LCs) are important antigen-presenting cells in the epidermis and may play a key role in the pathogenesis of psoriasis. It has been proven that LCs isolated from psoriatic lesions are abnormal. However, the mechanism of the abnormality has not been reported so far. OBJECTIVE: In the present study, we investigated the effect of psoriatic lesional keratinocytes on the maturation of LCs. METHODS: Monocytes isolated from healthy peripheral blood could differentiate into LCs in the presence of granulocyte-macrophage colony stimulating factor, interleukin (IL) 4 and transforming growth factor beta1 for 5 days. Then, human monocyte-derived LCs were cultured with supernatants from psoriatic lesional keratinocytes for another 2 days. Their phenotypes and phagocytic capacity were analyzed by flow cytometry. IL-12 secreted by LCs was determined by ELISA. RESULTS: Supernatants from psoriatic lesional keratinocytes could up-regulate the expression of HLA-DR and CD86 on LCs more significantly than supernatants from healthy keratinocytes, but less powerfully than lipopolysaccharide. The levels of IL-12 secreted by LCs also increased. In contrast, the expression of CD1a on LCs and their phagocytic capacity were reduced. CONCLUSION: Human monocyte-derived LCs cultured with supernatants from psoriatic lesional keratinocytes displayed the characteristics of maturation. This suggests that psoriatic lesional keratinocytes might secrete some factors that could promote the maturation of LCs, which may play important roles in immune reactions related to psoriasis. PMID- 11937733 TI - In situ localization of CD83-positive dendritic cells in psoriatic lesions. AB - BACKGROUND: Dendritic cells (DC) are considered to be the most potent antigen presenting cells, and CD83 is expressed at a high level on immune-competent, activated and mature DC. Although changes in the number or localization of mature and activated CD83+ DC could be expected in psoriasis, there is little information on such changes. AIM: Morphological identification of CD83+ DC in psoriatic skin lesions. MATERIALS AND METHODS: Immunohistochemical staining was performed in 5 specimens of psoriasis vulgaris and 6 specimens of pustular psoriasis. Formalin-fixed, paraffin-embedded sections were used for examination in this study. The skin sections were pretreated with 0.1% trypsin for 60 min at 37 degrees C prior to immunostaining for CD83. RESULTS: A small but significant subpopulation of CD83+ DC was found in the upper dermis. In addition, CD83+ DC were occasionally scattered in the epidermis. The most common distribution pattern of CD83+ DC was as clusters with mononuclear lymphoid cells in the upper dermis. CD83+ DC were in close contact with lymphocytes. High-intensity staining of CD83 antigens was detected not only on the surface, but also in the cytoplasm of DC. CONCLUSION: These results indicate that activated and mature CD83+ DC may play a role in the immune response in psoriasis and provide in vivo support for the concept that CD83+ DC provide signals for direct intralesional T cell activation. PMID- 11937734 TI - Lymphocytes treated by extracorporeal photopheresis demonstrate a drop in the Bcl 2/Bax ratio: a possible mechanism involved in extracorporeal-photopheresis induced apoptosis. AB - BACKGROUND: Recently, apoptosis has been identified in treated lymphocytes, prior to their re-infusion, when tested ex vivo. Previous work has demonstrated a close association between the genes p53, Bcl-2 and Bax and apoptosis induced by UV irradiation. OBJECTIVES: We wanted to establish whether the expression of the protein product of these genes was altered in lymphocytes treated with extracorporeal photopheresis (ECP) prior to re-infusion and therefore possibly implicated in the early apoptosis observed. METHOD: Lymphocytes were isolated immediately before treatment and immediately prior to re-infusion and tested for intracellular levels of p53, Bcl-2 and Bax proteins. RESULTS: No increase in p53 expression was observed at re-infusion; however, the mean fluorescent intensity ratio of the apoptotic inhibitor protein Bcl-2 to the apoptosis-inducing protein Bax dropped significantly. CONCLUSION: The early apoptosis observed in ECP treated lymphocytes at re-infusion might be attributed to dysregulation in the expression of the apoptotic genes Bcl-2 and Bax. PMID- 11937735 TI - Diagnostic features of pemphigus vulgaris in patients with bullous pemphigoid. Molecular analysis of autoantibody profile. AB - BACKGROUND: The simultaneous presence of features of pemphigus vulgaris (PV) in patients with bullous pemphigoid (BP) has previously been reported in the literature. OBJECTIVE: The purpose of this retrospective study is to present 13 patients with an initial diagnosis of BP, who subsequently demonstrated coexistent serological features of both BP and PV. METHODS: The following information on each patient was documented, at the time of initial diagnosis: clinical profile on presentation, histology, direct immunofluorescence, indirect immunofluorescence (IIF) using monkey esophagus as substrate, salt-split skin (SSS) and an immunoblot assay. Since all 13 patients failed to respond to conventional systemic therapy, intravenous immunoglobulin (IVIg) was used as an alternative treatment modality. Prior to initiating IVIg therapy, in all 13 patients, serological studies were performed. In addition to IIF using monkey esophagus, an immunoblot assay and SSS, an enzyme-linked immunosorbent assay (ELISA) was performed to detect antibodies to desmogleins. These different assays were done to identify pathological autoantibodies typical of BP and PV. A control group of 25 healthy normal individuals, 37 patients with BP, 17 patients with PV and 12 patients with pemphigus foliaceus were used for comparison of serological studies. RESULTS: At the time of initial presentation, histological and immunopathological studies confirmed the diagnosis of BP in all 13 patients. Prior to the initiation of IVIg therapy, results of IIF using monkey esophagus as substrate demonstrated high levels of anti-intercellular cement substance (anti ICS) or antikeratinocyte cell surface antibody. Sera of all 13 patients on SSS bound to the epidermal side of the split. In an immunoblot, using bovine gingival lysate as substrate, sera of 6 patients bound to both a 230-kD (BP Ag1) and 180 kD protein (BP Ag2), while 7 sera bound to only a 230-kD protein. All 13 patients had high levels of antibodies to desmoglein 3 on ELISA. In a pilot experiment, the anti-ICS antibody in sera from 6 random patients was found to be predominantly of the IgG4 subclass. Use of IVIg resulted in an effective clinical response and the maintenance of a prolonged clinical remission. CONCLUSION: In patients with BP, who are nonresponsive to conventional therapy, the presence of two autoimmune diseases or a dual diagnosis should be considered. PMID- 11937736 TI - Cell proliferation and lipid formation in hamster sebaceous gland cells. AB - BACKGROUND: The development of sebocytes and lipogenesis are known to be dependent on androgens. However, it is not fully understood whether growth factors are involved in the regulation of cell proliferation and lipid formation in sebaceous glands. OBJECTIVE: This study was designed to evaluate the effect of growth factors such as epidermal growth factor (EGF), transforming growth factor alpha (TGF-alpha), basic fibroblast growth factor (bFGF) and keratinocyte growth factor (KGF) on cell proliferation and lipogenesis in cultured hamster sebocytes. METHODS: Cell proliferation and intracellular lipid accumulation in these hamster sebocytes treated with growth factors were examined. RESULTS: EGF, TGF-alpha and bFGF augmented the proliferation of hamster sebocytes for 8 days in a time- and dose-dependent manner. However, KGF had no effect on their proliferation. Moreover, the accumulation of intracellular lipids consisting mainly of triglycerides was suppressed in EGF-, TGF-alpha-, bFGF- and KGF-treated hamster sebocytes. CONCLUSION: EGF, TGF-alpha and bFGF, but not KGF, have mitogenic activity on hamster sebocytes, and all these growth factors act as antilipogenic factors. Moreover, it is likely that the formation of intracellular lipid droplets is independent of cell proliferation in hamster sebocytes. PMID- 11937737 TI - Ichthyosiform mycosis fungoides. AB - BACKGROUND: Mycosis fungoides (MF) is a skin malignancy of T helper lymphocytes with a wide clinical spectrum. Among the atypical variants of MF, there is an ichthyosis-like presentation. However, to date, only 1 case of ichthyosiform MF has been reported. OBJECTIVE: Our goal was to summarize the clinical characteristics and course, and the pathological, immunohistochemical and molecular genetic findings on 4 patients with ichthyosiform MF. METHODS: A retrospective study was conducted. RESULTS: The 4 patients represented 1.8% of the 221 patients with MF seen by us since 1975. None progressed to systemic disease in up to 12 years (median, 10 years) after the onset of the cutaneous manifestations. Interestingly, skin lesions typical of so-called follicular MF (FMF) were associated in 3 of 4 cases, whereas cutaneous manifestations of classic MF were absent in all 4 patients. CONCLUSION: Ichthyosiform MF represents a rare variant within the clinicopathologic spectrum of MF usually featuring a benign course and a tendency to be associated with lesions of FMF but not with lesions of classic MF. PMID- 11937738 TI - Efficacy and safety of intralesional 5-fluorouracil in the treatment of keloids. AB - BACKGROUND: The treatment of keloids remains challenging. Cryosurgery and intralesional corticosteroids have been considered as the mainstream of therapy; however, the long-term use of corticosteroids has been found to be associated with serious side effects. Intralesional 5-fluorouracil (5-FU) has only been used in one study for the treatment of hypertrophic scars and keloids, mostly in combination with other treatments. The efficacy of 5-FU as an individual therapeutic agent is unknown. OBJECTIVE: To evaluate the efficacy and safety of intralesional injections of 5-FU in the treatment of small keloid lesions. METHODS: Twenty-four (12 male, 12 female) consecutive patients with keloids of 6 cm or less in their maximum dimension were treated with intralesional injections of 50-150 mg 5-FU per week for a maximum of 16 injections. RESULTS: One third (8/24, 33.3%) of the patients showed more than 75% flattening of the keloid. Three out of 8 patients (with >75% flattening) required less than 16 (13, 13 and 15) injections for achieving the desired response. Overall, about half of the patients showed more than 50% flattening of the treated keloid. A correlation with the duration of keloid was found. Six (54.5%) out of 11 patients with keloids of < or =5 years duration, in contrast to only 2 (15.4%) out of 13 patients with keloids of >5 years duration showed more than 75% flattening (p < 0.05). Side effects included pain (all patients), hyperpigmentation (all patients) and ulceration (1 patient). No difference in peripheral blood count was noted before, during and after the therapy. CONCLUSION: Intralesional 5-FU can be safely used for the management of small keloids of shorter duration. PMID- 11937739 TI - Lichen amyloidosus associated with Kimura's disease: successful treatment with cyclosporine. AB - We describe here a case of a 33-year-old man who had lichen amyloidosus associated with Kimura's disease. In this case, treatment with cyclosporine dramatically improved the lesions of both Kimura's disease and lichen amyloidosus. Although Kimura's disease and lichen amyloidosus are both rare distinct entities, to our knowledge, 11 cases of association of Kimura's disease and lichen amyloidosus have been described previously. PMID- 11937740 TI - Eccrine squamous syringometaplasia and syringomatous hyperplasia in association with linear scleroderma. AB - It is now well recognized that syringomatous hyperplasia with squamous syringometaplasia is a type of pseudoepitheliomatous hyperplasia responding to a variety of stimuli. We report a case of linear scleroderma with this process. Histopathology of the lesion showed numerous solid and cystic epithelial structures with squamous metaplasia within typical sclerodermatous skin lesions. Although the pathogenesis of the process remains unclear, dermal-derived growth factors in scleroderma and/or the mechanical effect of sclerosis could stimulate the proliferative response of eccrine epithelium. To the best of our knowledge, the association of syringomatous hyperplasia with eccrine squamous syringometaplasia and linear scleroderma has not been described previously. PMID- 11937741 TI - Fosinopril as a possible pemphigus-inducing drug. AB - Fosinopril has recently been added to the angiotensin-converting enzyme inhibitors inducing pemphigus. The observation of a patient in whom pemphigus vulgaris (PV) worsened after taking fosinopril prompted us to study an experimental way to assess its responsibility. Slices of normal human skin (NHS) were simultaneously incubated for 2, 6, 12 and 24 h at 4 degrees C with progressively diluted fosinopril and captopril solutions and used as indirect immunofluorescence (IIF) substrates for 2 sera containing anti-desmoglein-3 (anti Dsg3) antibodies at a dilution of 1/160. With captopril, IIF was negative, irrespective of dilution and time of incubation. Only at 1/40,000 dilution was IIF positive. With fosinopril, IIF was negative for the 2- and 6-hour-long incubations but turned positive after 12 h and so remained with all other solutions and incubation times. IIF negativity with captopril suggests that anti Dsg3 antibodies contained in the PV sera were unable to find molecules in NHS to bind to. Captopril would therefore induce acantholysis by blocking the adhesion molecules. With fosinopril, instead, a partial block of the adhesion molecules was seen only with the very concentrated solution, unlikely to occur in vivo. Fosinopril, therefore, is probably unable to block the adhesion molecules in vivo. Our method might be used to verify the acantholytic properties of a drug. PMID- 11937742 TI - Basal cell carcinoma of the earlobe after auricular acupuncture. AB - The genesis of familial and sporadic basal cell carcinomas involves activation of the Sonic hedgehog signal transduction pathway. Other known factors for the development of basal cell carcinoma are ultraviolet exposure, X-rays, race, age, gender and decreased DNA repair capacity. We here report the case of a right earlobe sporadic basal cell carcinoma in a 65-year-old woman. This case is unusual because of its earlobe localization and its association with multiple auricular acupuncture treatments. This observation suggests a connection between local traumas, which occur in the course of acupuncture treatment and ear piercing, and the genesis of basal cell carcinoma. The incidence of minor adverse events associated with acupuncture is high, but serious events are uncommon. Acupuncture is not known to date for promoting the development of tumors. This connection remains to be elucidated. PMID- 11937743 TI - Primary cutaneous cryptococcosis in an immunocompetent host: case report and review of the literature. AB - We report a 46-year-old male who developed cellulitis of his third right finger after being injured with a metallic object. Cryptococcus neoformans var. neoformans was isolated from both the skin and the metallic object. Systemic cryptococcosis and immunosuppression was excluded by appropriate tests. Complete healing was achieved after 10 months of itraconazole 100 mg/12 h and surgical excision of the remaining lesion. Our literature review shows another 16 cases of primary cutaneous cryptococcosis in nonimmunocompromised hosts. The clinical, mycological and therapeutic characteristics of these patients are reviewed. PMID- 11937744 TI - Onychomycosis due to Exophiala jeanselmei. AB - BACKGROUND: Exophiala jeanselmei is a dematiaceous fungus that may cause invasive diseases, particularly among immunocompromised hosts. Most reports mention cutaneous or subcutaneous lesions, but no case of nail involvement due to this fungus has been reported until now. CASE REPORT: A 60-year-old man presented with hyperkeratosis and black coloration of the nails of the two thumbs and the two big toes of 4 years' duration. He was a renal transplant recipient and had been treated with prednisone and azathioprine. E. jeanselmei was present on direct examination, then isolated in cultures on repeated samplings from all pathologic nails. He was cured after 6 months of treatment with itraconazole. COMMENTARY: We describe the first case of nail infection due to E. jeanselmei. Itraconazole, which provides a broad spectrum of action on fungal species and achieves high levels of active substance in many tissues, including the nails, appeared to be efficient upon such a nail localization of E. jeanselmei. PMID- 11937745 TI - Isolated streptococcal balanoposthitis in an adult patient. PMID- 11937746 TI - Atrophie blanche associated with interferon-alfa adjuvant therapy for melanoma: a cutaneous side effect related to the procoagulant activity of interferon? PMID- 11937747 TI - A case of localized pemphigoid with loss of toenails. PMID- 11937748 TI - Response of severe psoriasis to infliximab. PMID- 11937749 TI - Dicoumarin-induced skin necrosis. PMID- 11937750 TI - No evidence of Y250X transferrin receptor type 2 mutation in patients with porphyria cutanea tarda. A study of 38 cases. PMID- 11937751 TI - Unusual T cell pseudolymphoma with features of so-called pseudolymphomatous folliculitis. PMID- 11937752 TI - Bullous pemphigoid associated with acute myocarditis. PMID- 11937753 TI - Global analysis of gene expression: methods, interpretation, and pitfalls. AB - Over the past 15 years, global analysis of mRNA expression has emerged as a powerful strategy for biological discovery. Using the power of parallel processing, robotics, and computer-based informatics, a number of high-throughput methods have been devised. These include DNA microarrays, serial analysis of gene expression, quantitative RT-PCR, differential-display RT-PCR, and massively parallel signature sequencing. Each of these methods has inherent advantages and disadvantages, often related to expense, technical difficulty, specificity, and reliability. Further, the ability to generate large data sets of gene expression has led to new challenges in bioinformatics. Nonetheless, this technological revolution is transforming disease classification, gene discovery, and our understanding of regulatory gene networks. PMID- 11937754 TI - Renal transcriptomes: segmental analysis of differential expression. AB - BACKGROUND/AIMS: Progress accomplished by complete genomes and cDNA-sequencing projects calls for methods that fully use these resources to study gene expression patterns in characterized cell populations. However, since the number of functional genes cannot be readily inferred from the genomic sequence, it is highly desirable to make use of methods enabling to study both known and unknown genes. METHODS: The method of serial analysis of gene expression provides short diagnostic cDNA tags without bias towards known genes. In addition, the frequency of each tag in the library conveys quantitative information on gene expression. A microassay was set-up to perform serial analysis of gene expression in minute samples such as those obtained by microdissecting nephron segments. RESULTS: Studies carried out in the thick ascending limb of Henle's loop and the collecting duct of the mouse kidney provided expression data for several thousand genes. Known markers were found appropriately enriched, and several of the thick ascending limb or collecting duct specific transcripts had no database match. CONCLUSIONS: The microassay for serial analysis of gene expression makes possible large-scale quantitative measurements of mRNA levels in nephron segments. The comprehensive picture generated by analyzing both known and unknown transcripts in defined cell populations should help to discover genes with dedicated functions. PMID- 11937755 TI - Generation of kidney transcriptomes using serial analysis of gene expression. AB - Chronic renal disease initiation and progression remain incompletely understood. Genomewide expression monitoring should clarify the mechanisms which cause progressive renal disease by determining how clusters of genes coordinately change their activity. Serial analysis of gene expression (SAGE) is a technique of expression profiling which permits simultaneous and quantitative analysis of 9 to 13-bp sequence tags that correspond to unique mRNAs. Key principles of the technique are use of PCR in a manner to minimize distortion and serial concatenation of tags which facilitates sequencing and permits identification of many expressed genes in a single cDNA molecule. Tags are extracted from many concatenated sequences, counted using software, and identified by comparison with existing gene databases. In aggregate, gene expression profiles generated from a tag library comprise a transcriptome which represents a comprehensive and quantitative profile of genes expressed at the time of analysis. These global snapshots of gene expression patterns can better define basic cell biology and provide insights into disease pathogenesis by simultaneously determining the net consequences of gene-gene and gene-environment interactions on expression of thousands of genes. Rather than applying a priori assumptions (i.e., hypothesis testing), transcriptome analysis is hypothesis generating and requires no prior knowledge of gene expression. SAGE kidney transcriptomes, from normal animals and animals with progressive kidney disease, are being produced and can be analyzed for novel pathogenetic mechanisms. The use of SAGE and other genomic and proteomic tools should result in a better understanding of kidney disease pathogenesis and in identification of new therapeutic targets. PMID- 11937756 TI - Applications for microarrays in renal biology and medicine. AB - Groundbreaking recent developments, such as the near completion of human and mouse genome sequencing efforts and the emergence of robust microarray (gene chip) technologies, enabling comprehensive analysis of transcriptomes, provide new opportunities of unprecedented scale for researchers of kidney biology and disease. Combined with advanced computational and mathematical approaches for microarray data analysis, microarray applications promise to revolutionize our understanding of molecular mechanisms of kidney development and renal pathogenesis. New knowledge in this field will facilitate new approaches for molecular diagnostics, drug discovery, and eventually "personalized" renal medicine. In this review, we outline current and future research applications of microarray and computational approaches in renal biology and disease. We describe basic steps in microarray data analysis and introduce advanced computational approaches to optimize data mining of vast microarray datasets. PMID- 11937757 TI - Genes and proteins in renal development. AB - Metanephric kidney development begins with the formation of the metanephrogenic mesenchyme; this event depends on the prior action in the intermediate mesoderm of transcription factors such as Lim-1, Pax-2, Eya-1, and Foxc-1. Once it has formed, the mesenchyme secretes GDNF; this induces the nearby wolffian duct to produce a ureteric bud which invades the metanephrogenic mesenchyme and begins to arborize. Ureteric bud development and branching depends on the transcription factor Emx-2, the GDNF-cRet and probably the HGF/cMet, signalling systems, and the intracellular regulatory molecules formin IV and timeless. Proteins of the BMP family modulate ureteric bud branching and keep bud development in step with that of other tissue types. Proteins and glycosaminoglycans of the matrix, and their receptors, and also required. The metanephrogenic mesenchyme has a default fate of apoptosis and is dissuaded from suicide by factors secreted from the bud such as TGF-alpha, TIMP-2, EGF, and FGF-2. Other factors such as LIF and TGFbeta2 cooperate with these to induce clumps of mesenchyme cells to differentiate into nephrons, while BMP-7 appears to lead them instead to form stroma. As nephrons form, they express critical transcription factors such as WT-1, Pax-2, and Hoxa11 and d11, condense, and secrete Wnt-4. Wnt-4 acts in an autocrine loop to stimulate its own synthesis and is required for cells to differentiate into epithelia; its action is antagonized by sFRP-1, secreted by stroma, but this antagonism is itself inhibited by sFRP-2 made by the developing nephron. This system probably acts both to limit the spread of Wnt-4's influence and also to commit responding cells to their epithelial fate. As nephrons mature, regions of them differentiate to perform specific physiological functions, a process that requires the proteins WT-1, Lmx-1b, Notch-2, Jagged-1, and Hnf-1. PMID- 11937758 TI - Gene expression profiling in silico: relative expression of candidate angiogenesis associated genes in renal cell carcinomas. AB - Recent advances in gene expression profiling have led to the development of comprehensive databases which can be queried in various manners. In the present report, we have taken a list of genes previously associated with angiogenesis, either in in vivo or in in vitro models, and queried a commercial database established by GeneLogic to determine the relative expression of these candidate genes in normal kidneys and in renal cell carcinomas (RCC). We identified a number of genes, including CXCR4, matrix metalloproteinase 9, thrombospondin 2, and vascular endothelial growth factor, that were highly expressed in RCC versus normal tissue. One gene, hevin, appears to be selectively upregulated in RCC in contrast to downregulation of this gene in lung and colon tumors. This approach provides a powerful means to identify potential markers of tumor vascularization. PMID- 11937759 TI - Gene responses to hyperglycaemia. AB - The rising tide of diabetic nephropathy (DN) is arguably the most challenging clinical problem confronting nephrologists at the beginning of the new century. Central to the development of novel diagnostic and therapeutic strategies will be the elucidation of the molecular events that drive this complex disease. In this review, we briefly discuss the major growth factors and cytokines identified as mediators of tissue injury in DN in the "pre-gene subtraction" era. We then highlight the remarkable array of new molecular players already identified with the introduction of gene subtraction techniques, such as differential-display PCR, suppression-subtractive hybridization, and nucleotide micro-arrays, and discuss the likely impact of these technologies going forward. Finally, we summarize the current knowledge on the cell signalling events triggered by high glucose levels that influence gene expression in DN and that represent additional therapeutic targets in this setting. PMID- 11937761 TI - Microarray studies of gene expression in circulating leukocytes in kidney diseases. AB - The molecular characterization of changes in mRNA expression in renal tissue during disease is hampered by the acquisition of sufficient mRNA to do genomewide expression profiling. In many renal diseases, such as systemic lupus erythematosus, IgA nephropathy, antineutrophil cytoplasmic autoantibody (ANCA) associated glomerulonephritis, and small-vessel vasculitis (ANCA disease), circulating leukocytes play a role in onset, progression, and severity of the condition. Circulating leukocytes are readily isolated and supply sufficient mRNA for analysis, allowing molecular investigation into their involvement in the disease process. Our laboratory has undertaken a systematic study of the genomewide expression profiles of the circulating leukocytes from patients with a variety of renal diseases (ANCA disease, IgA nephropathy, lupus nephritis, focal segmental glomerulosclerosis), using the Affymetrix high-density gene chip array technology. Analysis of the data showed clustering of expressed genes unique for each individual disease group. These results imply that significant gene expression changes occur in leukocytes that are circulating in patients with renal diseases. In addition, gene expression has been studied in leukocytes activated in vitro by mechanisms that mimic pathogenic events in vivo. The expression levels of genes identified in in vitro studies were compared with the patient leukocyte gene expression to determine whether similar pathological events were occurring in vivo. PMID- 11937760 TI - Renal gene expression in embryonic and newborn diabetic mice. AB - Several novel genes that are upregulated in diabetic kidneys have been identified. Recently, transforming growth factor beta driven secreted proteins, i.e., connective tissue growth factor and gremlin (bone morphogenetic protein 2), have been identified, and their expression has been correlated with the tissue changes seen in diabetic nephropathy in the adult population. However, there are very few studies reported in the literature that describe the gene expression in the diabetic state during embryonic and neonatal life. It is well known that exposure to glucose or its epimer, i.e., mannose, induces marked dysmorphogenesis of the embryonic metanephros in an organ culture system. These changes are associated with ATP depletion and marked apoptosis, suggesting an oxidant stress in the induction of dysmorphogenesis of the embryonic metanephros. In view of the glucose-induced changes in the fetal metanephros, a diabetic state was induced by the administration of streptozotocin during pregnancy, and newborn mouse kidneys were processed for suppression subtractive hybridization-PCR. In addition, a diabetic state was induced in newborn diabetic mice, and after 1 week their kidneys were harvested and subjected to representational difference analysis of cDNA. Four novel genes with upregulated mRNA expression were identified. They included: (1) a translocase inner mitochondrial membrane 44 that is involved in the ATP-dependent import of preproteins from the cytosol into the mitochondrial matrix; (2) a kidney-specific aldo-keto reductase that utilizes NADPH and NADH as cofactors in the reduction of aromatic aldehydes and aldohexoses; (3) Rap1b, a Ras-related small GTP-binding protein that behaves as a GTPase and cycles between GTP-bound (active) and GDP-bound (inactive) states associated with conformational change, and (4) a fusion protein of ubiquitin polypeptide and ribosomal protein L40 (UbA(52) or ubiquitin/60) that is intimately involved in the ubiquitin dependent proteasome pathway related to the accelerated degradation of proteins under various stress conditions, such as those seen in patients with cancer and diabetes mellitus. PMID- 11937762 TI - Genes involved in atherosclerosis. AB - Atherosclerosis is a multifactorial disease that involves several genes and proteins. The purpose of this article is to focus on the arterial wall and to review lipoprotein receptors, growth factors, cytokines, chemokines, matrix metalloproteinases, adhesion molecules, and apoptosis genes and their involvement in atherogenesis. PMID- 11937763 TI - Endothelial gene responses to homocysteine: relation to atherosclerosis. AB - Chronic renal failure is associated with a 20-fold increased risk of cardiovascular mortality, in part due to accelerated atherosclerosis. Hyperhomocysteinemia (HHCy) is common in patients with chronic renal failure. It has been established that HHCy is an independent and graded risk factor for atherosclerosis, though the mechanisms responsible for it remain obscure. Using the cardiovascular cDNA microarray approach, we screened the expression of 600 cardiovascular relevant genes in human umbilical endothelial cells and identified a number of homocysteine-modulated genes. These differentially displayed genes were classified according to the functional outcome of their encoded proteins: endothelial motility cluster, signaling cluster, and lipid metabolism cluster. The results may be relevant to understanding the mechanisms underlying the pathophysiological effects of HHCy, particularly those leading to endothelial dysfunction and the pathogenesis of atherosclerosis. PMID- 11937764 TI - Genital herpes simplex virus type 1 infection: new fields for an old acquaintance? AB - Herpes simplex virus (HSV)-2 is the principal agent of chronic remittent genital herpes. Worldwide, only 10-20% of genital isolates are HSV-1. Studies from the British Isles and Scandinavia indicate, however, that HSV-1 is responsible for a significant proportion or even the majority of first clinical episodes of genital herpes in young women. Actual data show that a trend towards genital HSV-1 infections may also hold true for Germany. This article summarizes possible reasons for and consequences of the observed changes in the epidemiology of genital HSV infections. PMID- 11937765 TI - Liver transplantation-associated de novo hepatitis B virus infection: application of molecular evolutionary analysis. AB - OBJECTIVE: De novo hepatitis B virus (HBV) infection after liver transplantation has recently been reported to be associated with donors without serum hepatitis B surface antigen (HbsAg) but with hepatitis B core antigen (anti-HBc). We elucidate the source of de novo HBV infection after liver transplantation by molecular evolutionary analysis. METHODS: The serum sample was obtained from a recipient who underwent living related liver transplantation. He was negative for all HBV-related serum markers before the transplantation. The recipient became seropositive for HBsAg at 6 months after transplantation. The liver tissue was obtained from a donor who was seronegative for HBsAg, but positive for anti-HBs and anti-HBc. RESULTS: HBV DNA was detected from the serum and liver tissue in a recipient and donor, respectively. A total of 5 clones each of small-S gene of HBV from the donor and recipient were sequenced. A phylogenetic tree analysis based on small-S gene revealed that all isolates derived from the recipient and donor were clustered together within a close range of evolutionary distances. These results indicated that HBV was transmitted by the liver graft from the donor. CONCLUSIONS: Molecular evolutionary analysis can be adopted for the study of the transmission route of viral infection via organ transplantation. PMID- 11937766 TI - Analysis of background factors influencing long-term prognosis of patients with chronic hepatitis C treated with interferon. AB - OBJECTIVE: Interferon (IFN) therapy has been used as antiviral therapy for chronic hepatitis C (CH-C); however, complete response to the therapy is observed in only about 30% of patients in Japan. Background factors involved in the responsiveness to IFN therapy, and progression to liver cirrhosis (LC) and hepatocellular carcinoma (HCC) after IFN therapy have not yet been sufficiently investigated. METHODS: One hundred twenty-one patients with CH-C who received IFN therapy at Showa University Hospital between 1984 and 1999 were analyzed. RESULTS: At 6 months after the termination of IFN therapy, 53 patients achieved a complete response, 11 patients incomplete response, and 57 patients no response. During a mean follow-up of 52.7 months, 12 patients progressed to LC, and 10 patients developed HCC. Multivariate analysis showed that significant independent factors involved in progression to LC were platelet count and the efficacy of IFN therapy. The significant independent factor involved in the development of HCC was platelet count. The factor involved in the therapeutic effect at 6 months after the termination of IFN administration was the serum hepatitis C virus (HCV) RNA levels before IFN therapy. CONCLUSION: Patients with high HCV RNA levels and low platelet counts should be considered to be at high risk of progressing to LC and developing of HCC and should be carefully followed after IFN therapy using ultrasonography, CT scan and MRI. PMID- 11937767 TI - Clinical and subclinical reactivation of genital herpes virus. AB - Reactivations of herpes simplex virus (HSV) either symptomatically (recrudescence) or without symptoms (recurrence) are well documented. As an asymptomatic reactivation may contribute to transmitting HSV to potential acceptors the frequency of reactivations should be evaluated. In order to evaluate the frequency of HSV-2 reactivation 173 genital swabs of a group of women chosen at random were analyzed by nested PCR. 34 (19.6%) showed clinical evidence of a herpes infection, 77 (44.5%) had no symptoms at all and 62 (35.8%) had other symptoms. In 26 (15%) HSV-DNA was detected. 11 (38.4%) could be characterized as asymptomatic reactivations. Typing of the HSV-positive swabs resulted in 11 HSV-2 and 10 HSV-1 strains. Additionally 18 HSV-positive swabs of the oral cavity resulted in 15 (83.2%) HSV-1 and 3 (16.4%) HSV-2 strains. The results of typing indicate a change of HSV-1 and HSV-2 epidemiology. PMID- 11937768 TI - Evaluation of HBs, HBc, and frCP virus-like particles for expression of human papillomavirus 16 E7 oncoprotein epitopes. AB - OBJECTIVES: In an attempt to develop virus-like particles (VLPs) as experimental vaccine against human papilloma virus (HPV)-induced tumours, the HPV16 E7 oncoprotein epitopes spanning amino acid (aa) residues 35-98 were expressed on three proteins capable of VLP formation: hepatitis B virus (HBV) surface (HBs) and core (HBc) antigens, and RNA phage fr coats (frCP). METHODS: The profile of immunoglobulin isotypes induced in Balb/C mice after immunization with purified chimeric proteins was studied. RESULTS: The HBs*-E7(35-54) protein expressing E7 residues 35-54 between residues 139 and 142 of the HBs carrier formed HBs-like particles in Saccharomyces cerevisiae. The HBc Delta-E7(35-98), but not the frCP E7(35-98), ensured VLP formation in Escherichia coli. In Balb/C mice, the HBs* E7(35-54) VLPs predominantly induced an anti-E7 antibody, but not anti-HBs carrier response, whereas the HBc Delta-E7(35-98) VLPs induced a lower anti-E7 compared to anti-HBc carrier response. The frCP-E7(35-98) protein elicited equally high antibody responses to both E7 and frCP carrier. Analysis of the immunoglobulin G isotype profile of the antibodies induced by the E7-carrying chimeras showed that the HBs and frCP derivatives were capable of eliciting the Th1 and Th2 subsets of T helper cells, whereas the HBc-derived chimeras elicited only the Th2 subset. CONCLUSIONS: The HBs and HBc, but not frCP carriers support an efficient outcome for VLPs carrying the HPV16 E7 epitopes. All chimeric proteins may be regarded as potential vaccine candidates. PMID- 11937769 TI - Viral dynamics and pharmacokinetics in combined interferon alfa-2b and ribavirin therapy for patients infected with hepatitis C virus of genotype 1b and high pretreatment viral load. AB - OBJECTIVE: The aim of this study was to clarify the viral dynamics and single- and multiple-dose pharmacokinetics of ribavirin and interferon (IFN) alfa-2b in the virologic response to combination therapy with both compounds in patients infected with hepatitis C virus (HCV) genotype 1b and high viral load. METHODS: Fourteen patients received high-dose daily induction therapy followed by intermittent maintenance therapy with IFN alfa-2b and daily oral ribavirin for 24 weeks, and followed up for 24 weeks after treatment. Single- and multiple-dose pharmacokinetic studies and viral dynamics were assessed by serial measurements of serum concentrations of both compounds and HCV RNA, respectively, at weeks 1 and 24. RESULTS: During treatment, all 14 patients showed biochemical response (i.e., normalization of serum alanine transaminase activity), while 11 showed virologic response (i.e., undetectable serum HCV RNA level by qualitative polymerase chain reaction assay). Sustained biochemical and virologic response after cessation of treatment was noted in 8 and 2 patients, respectively. Serum ribavirin concentrations asymptoted by 4 weeks of treatment. Serum ribavirin concentrations in steady state, and maximum concentration and accumulation rate of ribavirin after multiple dosing were significantly higher in the presence of sustained virologic response. HCV-related parameters were not significantly associated with sustained virologic response. CONCLUSIONS: Continuous exposure and tissue accumulation of ribavirin may be necessary for sustained virologic response to combination therapy in chronic hepatitis C with genotype 1b and high viral load. Pharmacokinetic analysis of ribavirin provides information on its mechanism of action and for developing more rational treatment for IFN-resistant HCV. PMID- 11937770 TI - Respiratory syncytial virus enhances the expression of CD11b molecules and the generation of superoxide anion by human eosinophils primed with platelet activating factor. AB - BACKGROUND: Human respiratory syncytial virus (RSV) infection in infancy and early childhood causes acute bronchiolitis and exacerbates bronchial asthma. Eosinophil infiltration may contribute to airway obstruction in RSV infection. OBJECTIVE: We hypothesized that RSV affects eosinophil function. METHODS: Eosinophil activation was evaluated by chemiluminescent detection of superoxide anion (O(2)(-)) generation. Expression of CD11b on eosinophils was determined by flow cytometry. RESULTS: Although RSV did not induce O(2)(-) generation by resting eosinophils, RSV enhanced O(2)(-) generation of eosinophils primed with platelet-activating factor (PAF). Enhancement was significantly inhibited by either continuous agitation to prevent eosinophil adhesion to test tube surfaces or by pretreating cells with anti-CD18 antibody, suggesting that the stimulatory effects of RSV on eosinophils depend on cell adhesion via beta(2)-integrins. In fact, RSV enhanced PAF-induced CD11b expression by eosinophils. CONCLUSIONS These findings suggest that RSV enhances eosinophil CD11b expression and O(2)(-) generation induced by PAF. Thus, RSV infection may exacerbate airway inflammation by enhancing mediator release from eosinophils. PMID- 11937771 TI - Distinct mutational drug resistance profiles of HIV-1 RNA in plasma and culture isolates of patients receiving antiretroviral therapy. AB - The differences in profiles of HIV mutational drug resistance between paired plasma samples and culture isolates were analyzed in treated and untreated HIV infected patients. The DNA sequence of the entire protease gene and codons 1-320 of the reverse transcriptase gene was analyzed from the paired plasma samples and viral isolates. Differences in the distribution of resistance mutations from the two sources were detected in 13 of 14 patients receiving antiretroviral therapy. Some excess mutations, observed in plasma samples but not in culture isolates, are reportedly associated with resistance to antiretroviral drugs. These results emphasize the need for standardization and clinical laboratory use of phenotypic assays that avoid extensive cocultivation. PMID- 11937772 TI - Profound sex-specific effects on incubation times for transmission of bovine spongiform encephalopathy to mice. AB - Four strains of mice were inoculated intracerebrally with a primary isolate of bovine spongiform encephalopathy (BSE) and the cloned mouse-adapted scrapie strain ME7. Clinical prion disease diagnosis was made at the appearance of three or more neurological symptoms and their persistence for 3 consecutive weeks and confirmed by neuropathological criteria. For BSE, incubation periods were profoundly different between the sexes in all four mouse strains, being longer in the females. In contrast, ME7 scrapie incubation times were similar between the sexes. Our results indicate that sex-specific processes are involved in the course of primary BSE transmission. Research into this phenomenon may provide clues to the prophylaxis of BSE and have possible implications for new variant Creutzfeldt-Jakob disease in humans. PMID- 11937773 TI - Mapping of epitopes on the fiber knobs of human adenovirus serotypes 8 and 15. AB - In order to obtain information on the linear antigenic epitopes on fiber knobs of adenovirus serotypes 8 (Ad8) and 15 (Ad15) of subgenus D, the binding of polyclonal virus- and fiber-specific rabbit antibodies to overlapping peptides covering the fiber knob was studied. The main antigenic epitopes of the fiber knob of Ad8 (FK8) were represented by the peptides P4 [amino acids (aa) 213-227], P6 (aa 233-247), P11 (aa 283-297), P13 (aa 303-317) and P15 (aa 316-325); the peptides P1 (aa 183-197), P8 (aa 253-267), P10 (aa 273-287) and P23 (aa 340-349) were moderately reactive. The peptides P4, P6, P11, P13 and P15 span the beta strands C, D, G and H, parts of the CD and DG loops and the complete GH loop of the fiber knob. The main epitopes of the fiber knob of Ad15 (FK15) were represented by peptides P5 (aa 198-212), P10 (aa 223-237), P12 (aa 233-247), P13 (aa 238-252), P26 (aa 303-317), P29 (aa 318-332) and P32 (aa 333-347), spanning the beta strands B, D, G, H and I, partly strand C, the CD loop, parts of the AB and DG loops, the GH and HI loops and the N-terminal part of the IJ loop. The amino acid sequence alignment showed that the location of the linear FK8 and FK15 epitopes was found to be overlapping to a major extent. Two serotype-specific epitopes were determined on FK15, represented by P10 and P13. PMID- 11937774 TI - A rational approach to the selection of useful drugs for clinical practice. AB - Confusion exists concerning optimal selection of drugs for clinical practice. Many reasons contribute to this confusion which derives from various sources including industry, the FDA and often less than ideal education of physicians in clinical pharmacology during medical school and thereafter. This presentation does not focus on allocating blame for the current unsatisfactory situation, but rather offers a solution to improve drug therapy. Our educational solution consists of a paradigm for rational drug therapy, specifically a checklist of eight criteria for physicians to review before prescribing drugs. Such a review would facilitate better estimation by physicians of risk/benefit ratios involving new, often expensive and, in some cases, questionably effective and safe drugs. PMID- 11937775 TI - Effects of STA(2), a thromboxane A(2) mimetic, in inducing airflow obstruction and airway microvascular leakage in guinea pigs. AB - BACKGROUND: U-46619, a thromboxane A(2) (TXA(2)) mimetic, is shown to cause airway microvascular leakage, although the effects is weak when comparing with that to induce bronchoconstriction in guinea pigs. OBJECTIVE: In order to know the airway effect of TXA(2) more accurately, we have examined the effects of STA(2), a TXA(2) mimetic with higher affinity to TXA(2) (TP) receptors than U 46619, to induce airway microvascular leakage and airflow obstruction. METHODS: Anesthetized and ventilated guinea pigs were i.v. given STA(2) (3-30 nmol/kg) or U-46619 (3-100 nmol/kg) 1 min after i.v. Evans blue dye. STA(2)- and U-46619 induced increases in lung resistance (R(L)) was measured for 6 min. The amount of extravasated Evans blue dye in the lower airways was, then, examined as an index of leakage. In selected animals, specific TP receptor antagonists (10 microg/kg S 1452 or 10 mg/kg ONO-3708) were pretreated i.v. RESULTS: Both STA(2) and U-46619 induced significant increases in leakage and airflow obstruction. However, STA(2) induced a slow and significantly less increase in R(L) but caused a significantly greater increase in extravasation of Evans blue dye compared to U-46619. Specific TP receptor antagonists completely abolished both airway effects induced by STA(2) and U-46619. CONCLUSION: Our present results have supported a possibility that TXA(2) induces microvascular leakage as well as bronchoconstriction in the airways. PMID- 11937776 TI - Binding and functional affinity of sarpogrelate, its metabolite m-1 and ketanserin for human recombinant alpha-1-adrenoceptor subtypes. AB - Serotonin (5-HT(2)) antagonists show high affinity for the alpha(1)-adrenoceptor (alpha(1)-AR) in addition to the 5-HT(2) receptor. In the present study we compared the pharmacological characteristics of a new 5-HT(2) antagonist sarpogrelate and its active metabolite M-1 with those of ketanserin on human recombinant alpha(1)-AR subtypes. In the binding study, sarpogrelate, M-1 and ketanserin produced concentration-dependent inhibition of (3)H-prazosin binding to alpha(1)-ARs. Among the three drugs, ketanserin showed the highest affinity for alpha(1a)-, alpha(1b)- and alpha(1d)-ARs (pKi 8.0, 8.3 and 7.6, respectively). Sarpogrelate had a relatively low affinity for the three subtypes (6.3, 6.4 and 6.3, respectively) and M-1 showed medium affinity (7.1, 7.1 and 6.1, respectively). Chinese hamster ovary (CHO) cells expressing each alpha(1)-AR subtype showed concentration-dependent inositol phosphate (IP) accumulation in response to phenylephrine. The concentration response curves were shifted to the right by three drugs, and the pKb values were close to the pKi values in the binding study. In addition to these effects, sarpogrelate and M-1, but not ketanserin produced an increase in the basal IP level of alpha(1d)-expressed CHO cells, although the increase was less than that of phenylephrine. The present results indicate that sarpogrelate and M-1 have antagonistic activity to the three alpha(1)-AR subtypes, but their affinities are significantly lower than those of ketanserin. PMID- 11937778 TI - In vivo and in vitro effects of bromelain on PGE(2) and SP concentrations in the inflammatory exudate in rats. AB - The effects of bromelain were examined in rats with subcutaneous carrageenin induced inflammation. After oral in vivo administration, bromelain (10 and 20 mg/kg p.o.) induced a significant decrease of both PGE(2) and substance P concentrations in the exudate. When added to the inflammatory exudate in vitro, the drug (25, 50, 100 microg/ml) did not affect PGE(2) concentrations and induced an increase in the substance P levels. Our data indicate that bromelain reduces the production of two key mediators of inflammation. This effect does not seem to be related to a direct action of the drug on PGE(2) and SP released in the exudate in response to the inflammatory stimulus. PMID- 11937777 TI - In vitro and in vivo activities of three acridine thioethers against Leishmania donovani. AB - The lack of definitive chemotherapeutic agents to fight against visceral leishmaniasis has lead to the testing of numerous compounds. In the present work, we carry out an in-depth study of the activity against Leishmania donovani of three acridine derivatives both in vitro and in vivo. These compounds have proven to be highly effective at medium and high concentrations of 10 microg/ml, against both flagellate and nonflagellate forms of the parasite, which, though obtained in vitro, closely resemble natural intracellular amastigotes. The in vivo assays showed a significant reduction in the percentage of parasitation versus control, for all the compounds tested. In addition, we have studied the possible mechanism by which these acridine derivatives act: they displayed a greater inhibitory effect against macromolecule synthesis in treated flagellates, yet alterations are also caused in the production of end metabolites and in the activity of different enzymes. The data obtained indicate that the acridine derivatives had several targets, one of them is the synthesis of nucleic acids and proteins, while the second one might be interaction with the carbohydrate and energy production processes in the parasite. This conclusion is consistent with our observations concerning the ultrastructural changes induced in the parasite by these compounds principally at the mitochondrial level. PMID- 11937779 TI - Diltiazem reduces apoptosis in rat hepatocytes subjected to warm hypoxia reoxygenation. AB - Interruption of hepatic blood flow is necessary in surgery, but the liver is sensitive to ischemia and reperfusion. Hypoxia induces an increase in intracellular calcium concentration. In previous studies, we have shown that hypoxia-reoxygenation (H/R) increased calcium influx and induced JNK(1)/SAPK(1) activation which was involved in the triggering of apoptosis. The aim of this study was to demonstrate that diltiazem, a calcium inhibitor, reduced JNK(1)/SAPK(1) activation and consequently could decrease H/R-induced apoptosis. Experiments were performed, in the presence of diltiazem, on primary cultured rat hepatocytes, subjected to warm H/R phases and in a liver ischemia-reperfusion model. The activation status of JNK(1)/SAPK(1) was evaluated by immunoprecipitation and immunohistolocalisation experiments, while apoptosis was assessed by measuring caspase activity and by TUNEL labeling. Diltiazem inhibited H/R-induced JNK(1)/SAPK(1) activation and decreased apoptosis. It could be used to improve postoperative liver function. PMID- 11937780 TI - Effect of YM-44781, YM-44778 and YM-49598, novel tachykinin antagonists, in a drug-induced bladder contraction model. AB - The radioligand binding profiles and in vivo pharmacological characteristics of YM-44781, YM-44778 and YM-49598, novel non-peptide tachykinin receptor antagonists, were examined and compared to those of FK-888 and GR-159897. Since no functional NK(3) receptors were found in the rat bladder, the emphasis will be on the other two subtypes. YM-44781 and YM-49598 exhibited high binding affinities at NK(2) (pK(i) = 9.94 +/- 0.03) and NK(1) (pK(i) = 9.09 +/- 0.02) receptors, respectively, whereas YM-44778 exhibited high binding affinities at both NK(1) (pK(i) = 8.08 +/- 0.07) and NK(2) (pK(i) = 8.55 + 0.04) receptors stably transfected in CHO-K1 cells (Chinese hamster ovary cells). In an in vivo rat model, a drug-induced bladder contraction model, antagonism of the contractions produced by the selective NK(2) receptor agonist, [betaAla8]neurokinin A (4-10) (10 microg x kg(-1) i.v.) was observed after intravenous administration (dose range 0.001-1 mg x kg(-1)) of YM-44781 and YM 44778 (IC(50) = 27 +/- 8 and 100 +/- 44 microg x kg(-1), respectively). YM-44781 was more potent (about 3-fold) than YM-44778. YM-49598 was almost inactive but produced a potent inhibition (IC(50) = 11 +/- 7 microg x kg(-1)) of the contraction of the rat urinary bladder induced by challenge with the NK(1) selective receptor agonist [Sar9,Met(O(2))11]substance P sulphone (0.3 microg x kg(-1)). YM-44781 and YM-44778 did not produce major inhibition of [Sar9,Met(O(2))11]substance P-induced bladder contraction. These findings indicate that YM-44781 and YM-49598 are potent NK(2) and NK(1) receptor antagonists, respectively, whereas YM-44778 is a nonselective NK(2)/NK(1) receptor antagonist in the drug-induced bladder contraction model. PMID- 11937781 TI - Antioxidant potential of a novel tetrapeptide derivative in isoproterenol-induced myocardial necrosis in rats. AB - A novel tetrapeptide derivative Boc-Lys(Boc)-Arg-Asp-Ser(tbu)-OtBu (PEP1261) has been tested in vivo in isoproterenol (ISO) hydrochloride (HCl)-induced myocardial necrosis in rats. ISO x HCl induces myocardial necrosis in rats which is accompanied by the distinct increase in heart weight, marked electrocardiographic changes, increase in the levels of serum marker enzymes and lipid peroxides and decrease in the levels of antioxidants. PEP1261 (5 mg/kg body weight i.p.) pre- and post-treatment effectively decreases serum marker enzyme levels, while the electrocardiographic changes get restored towards normalcy. PEP1261 also inhibits the action of the free radicals toxicity by increasing the levels of antioxidants and histological studies confirm the above findings. This study shows that PEP1261 could serve as an excellent cardioprotective agent possessing membrane stabilizing action. PMID- 11937782 TI - Effects of nipradilol on alpha-adrenoceptor function in ocular arteries. AB - The effects of nipradilol, a drug used in the treatment of glaucoma, on the contractions induced by noradrenaline and phenylephrine in isolated dog central retinal, external and internal ophthalmic arteries and pig ciliary arteries were investigated. In dog ocular arteries treated with oxyhemoglobin (1.6 x 10(-5) mol/l) to adsorb nitric oxide, noradrenaline (2 x 10(-8) to 10(-5) mol/l) produced a concentration-related contraction which was markedly inhibited by prazosin but not by yohimbine. Nipradilol (10(-9) to 10(-7) mol/l) slightly but significantly inhibited the noradrenaline-induced contraction in a concentration related manner, but the inhibitory potency and efficacy were much less than those of prazosin. However, nipradilol inhibited the phenylephrine-induced contraction with a similar PA(2) value of prazosin. In pig ciliary arteries treated with oxyhemoglobin, noradrenaline-induced contraction was slightly inhibited by prazosin but markedly inhibited by yohimbine. Nipradilol, similarly to timolol, did not inhibit, but rather tended to potentiate, the contraction elicited by noradrenaline. The contraction induced by phenylephrine was significantly inhibited by prazosin and nipradilol. It is concluded that nipradilol acts as an alpha(1)-adrenoceptor antagonist (but not as an alpha(2)-adrenoceptor antagonist) in the ocular arteries, which may partially explain its ocular-pressure-lowering mechanism. Taken together with the results of our previous studies, the potencies of the nipradilol-induced vascular actions in ocular arteries are found to be in the following order: beta-adrenoceptor inhibition > alpha(1)-adrenoceptor inhibition falling dots direct vasodilation via a release of nitric oxide. PMID- 11937861 TI - Bone: formation by autoinduction. 1965. PMID- 11937862 TI - Bioactive materials in orthopaedic surgery: overview and regulatory considerations. AB - Although bone graft continues to be the standard against which other skeletal substitutes are measured, orthopaedic surgeons soon will have various new tools available for skeletal reconstruction. With these tools, the distinctions between inert materials, resorbables, bioactive materials, transplantable tissues, engineered tissues, drugs, and composites become indistinct. Although almost any implanted material evokes some type of host reaction, in the context of reconstructive orthopaedic surgery, bioactive materials can be considered osteogenic, osteoconductive, osteoinductive, or a combination thereof. In the United States, the regulatory control of a new skeletal substitute material is complex, and is based in part on whether the material is considered primarily a biologic, a drug, or a medical device. Different agencies within the Food and Drug Administration have responsibility for regulatory control of different types of products. Although some new materials can be approved by a Premarket Notification (510(K)), others require a Premarket Approval Application. Regulations are being developed that affect the extent of regulatory influence over minimally manipulated tissues for transplantation. PMID- 11937863 TI - Use of injectable calcium phosphate cement for fracture fixation: a review. AB - Injectable osteoconductive calcium phosphate cements have been introduced as an adjunct to internal fixation for treating selected fractures. These cements harden without producing much heat, develop compressive strength, and are remodeled slowly in vivo. The main purpose of the cement is to fill voids in metaphyseal bone, thereby reducing the need for bone graft, but cements also may improve the holding strength around metal devices in osteoporotic bone. Several bioactive cements are being developed. One of these cements, Skeletal Repair System, is available in Europe and has been approved by the United States Food and Drug Administration for use in selected distal radius fractures. Cadaveric studies have shown that using Skeletal Repair System cement with conventional metal fixation in certain fractures of the distal radius, tibial plateau, proximal femur, and calcaneus can produce better stability, stiffness, and strength than metal fixation alone. Early clinical results have shown reduced time to full weightbearing when cement has been used for augmentation of tibial plateau and calcaneal fractures, more rapid gain of strength and range of motion when used in distal radius fractures, and improved stability in certain hip fractures. Bioactive cements in general also may prove useful in vertebroplasty. PMID- 11937864 TI - Osteoinductive molecules in orthopaedics: basic science and preclinical studies. AB - Osteoinductive molecules are characterized by their ability to promote the formation of bone. Most osteoinductive molecules are cytokines, which are extracellular proteins or peptides that mediate cell to cell signaling. Examples of osteoinductive cytokines are certain bone morphogenetic proteins and some growth and differentiation factors. Some osteoinductive molecules are not secreted molecules. LIM mineralization protein-1 is an example of an intracellular osteoinductive molecule. Significant advances have been made in characterizing the molecular composition and mechanism of action of these osteoinductive molecules. Preclinical studies with these molecules have provided better understanding of the doses, formulation, and delivery mechanism necessary for effective bone formation in model systems of spinal fusion and other orthopaedic problems. The current authors will review the most important basic science and preclinical studies involving these osteoinductive molecules. PMID- 11937865 TI - Nonallograft osteoconductive bone graft substitutes. AB - An estimated 500,000 to 600,000 bone grafting procedures are done annually in the United States. Approximately (1/2) of these surgeries involve spinal arthrodesis whereas 35% to 40% are used for general orthopaedic applications. Synthetic bone graft substitutes currently represent only 10% of the bone graft market, but their share is increasing as experience and confidence in their use are accrued. Despite 15 to 20 years of clinical experience with various synthetic substitutes, there have been few welldesigned, controlled clinical trials of these implants. Synthetic bone graft substitutes consist of hydroxyapatite, tricalcium phosphate, calcium sulfate, or a combination of these minerals. Their fabrication technique, crystallinity, pore dimensions, mechanical properties, and resorption rate vary. All synthetic porous substitutes share numerous advantages over autografts and allografts including their unlimited supply, easy sterilization, and storage. However, the degree to which the substitute provides an osteoconductive structural framework or matrix for new bone ingrowth differs among implants. Disadvantages of ceramic implants include brittle handling properties, variable rates of resorption, poor performance in diaphyseal defects, and potentially adverse effects on normal bone remodeling. These inherent weaknesses have refocused their primary use to bone graft extenders and carriers for pharmaceuticals. The composition, histologic features, indications, and clinical experience of several of the synthetic bone graft substitutes approved for orthopaedic use in the United States are reviewed. PMID- 11937866 TI - Osteoconductive coatings for total joint arthroplasty. AB - Osteoconductive calcium phosphate coatings for total joint arthroplasty have been in clinical use since the mid1980s. The basic principles involved and basic science evidence for the efficacy of osteoconductive coatings were examined. Hydroxyapatite coatings provide consistent and better filling with bone of the gaps and spaces around cementless joint components after surgery as compared with porous-coated implant surfaces, resulting in better implant stability. Of all the calcium phosphate coatings, hydroxyapatite coatings have had the most widespread application in hip arthroplasty. Their clinical advantages over more conventional implant surfaces are evident in primary and revision hip arthroplasties. A clinical survival rate in the author's series of 97% at a minimum of 11 years followup for the femoral component in a young active patient population (average age, 53 years) was obtained with no mechanical failures. The average polyethylene wear rate in this group was 0.129 mm/year. In a similar group of young patients with revision arthroplasty using hydroxyapatite-coated femoral components, an 11 year survival rate of 93% was obtained. Histologic analysis of specimens retrieved at autopsy confirmed the excellent bony fixation of components. Advantages of the more recent biomimetic hydroxyapatite coatings were examined. PMID- 11937867 TI - Connective tissue progenitors: practical concepts for clinical applications. AB - Tissue engineering can be defined as any effort to create or induce the formation of a specific tissue in a specific location through the selection and manipulation of cells, matrices, and biologic stimuli. The biologic concepts and the biochemical and biophysical principles on which these efforts are based have become an exciting and rapidly evolving field of biomedical research. More importantly, tissue engineering is becoming a clinical reality in the practice of orthopaedic surgery, providing patients and physicians with an expanding set of practical tools for effective therapy. New and improved matrices and bioactive factors inevitably will play important roles in the evolution of orthopaedic tissue engineering. However, tissue engineering never can stray far from fundamental biologic principles, and one of these is that cells do all the work. No new tissue forms except through the activity of living cells. No bone graft, no matrix, no growth factor, no cytokine can contribute to the generation or integration of new tissue, except through the influence it has on the behavior of cells. The efficacy of all current clinical tools depends entirely on the cells in the grafted site, particularly the small subset of stem cells and progenitor cells that are capable of generating new tissue. The current authors review a series of key biologic concepts related to the rational design and selection of composites of cells and matrices in contemporary bone grafting and tissue engineering efforts. The functional paradigms of stem cell biology are reviewed, including self renewal, asymmetric and symmetric mitosis, and lineage restriction. Several potential sources for autogenous stem cells for connective tissues are discussed. Finally, a simple mathematical model is introduced as a tool for understanding the functional demands placed on stem cells and progenitors in a graft site and to provide a conceptual framework for the rational design of cell matrix composite grafts. PMID- 11937868 TI - Properties of osteoconductive biomaterials: calcium phosphates. AB - Bone is formed by a series of complex events involving the mineralization of extracellular matrix proteins rigidly orchestrated by cells with specific functions of maintaining the integrity of the bone. Bone, similar to other calcified tissues, is an intimate composite of the organic (collagen and noncollagenous proteins) and inorganic or mineral phases. The bone mineral idealized as calcium hydroxyapatite, Ca10 (PO4)(6)(OH)2, is a carbonatehydroxyapatite, approximated by the formula: (Ca,X)(10)(PO4,HPO4,CO3)(6)(OH,Y)2, where X are cations (magnesium, sodium, strontium ions) that can substitute for the calcium ions, and Y are anions (chloride or fluoride ions) that can substitute for the hydroxyl group. The current author presents a brief review of CaP biomaterials that now are used as grafts for bone repair, augmentation, or substitution. Commercially-available CaP biomaterials differ in origin (natural or synthetic), composition (hydroxyapatite, beta-tricalcium phosphate, and biphasic CaP), or physical forms (particulates, blocks, cements, coatings on metal implants, composites with polymers), and in physicochemical properties. CaP biomaterials have outstanding properties: similarity in composition to bone mineral; bioactivity (ability to form bone apatitelike material or carbonate hydroxyapatite on their surfaces), ability to promote cellular function and expression leading to formation of a uniquely strong bone-CaP biomaterial interface; and osteoconductivity (ability to provide the appropriate scaffold or template for bone formation). In addition, CaP biomaterials with appropriate three-dimensional geometry are able to bind and concentrate endogenous bone morphogenetic proteins in circulation, and may become osteoinductive (capable of osteogenesis), and can be effective carriers of bone cell seeds. Therefore, CaP biomaterials potentially are useful in tissue engineering for regeneration of hard tissues. PMID- 11937869 TI - Demineralized bone matrix and native bone morphogenetic protein in orthopaedic surgery. AB - The recognition that demineralized bone matrix could induce bone formation when placed in mammalian skeletal muscle led to preclinical studies of crude native insoluble bone morphogenetic protein and noncollagenous protein, followed by the clinical application of demineralized bone matrix, chemosterilized autolyzed antigen-extracted allogenic bone, and autolyzed antigen-extracted allogenic bone matrix gelatin. Cultural norms and regulatory agencies influence the availability of different demineralized bone matrix preparations in different parts of the world, but there is continued interest in the biologic structure of native insoluble bone morphogenetic protein and noncollagenous protein aggregates and the applied science of osteoinduction and osteoconduction in reconstructive orthopaedic surgery. Demineralized bone matrix is not widely available in Asia, but tissue processing facilities in the United States distribute demineralized bone matrix materials with different carriers, handling properties, and possibly osteoinductive potential. The purpose of the current study was to review the development and use of various preparations of demineralized bone matrix materials. PMID- 11937870 TI - Clinical evaluation of recombinant human bone morphogenetic protein-2. AB - Recombinant human bone morphogenetic protein-2 is an osteoinductive protein that plays a pivotal role in bone growth and regeneration. Several hundred studies were conducted in the past 7 years in numerous animal models to establish unequivocally the efficacy, safety, mechanism of action, pharmacokinetics, and surgical handling properties of recombinant human bone morphogenetic protein-2, building a solid foundation for clinical development programs. Pilot clinical trials have shown the feasibility and safety of recombinant human bone morphogenetic protein-2 treatment, and defined the effective dose for its use in open long bone fractures and for augmentation or preservation of the alveolar bone in the dental ridge. Prospective observational clinical studies helped define clinical efficacy end points, identify significant variables, and estimate appropriate population sample size for pivotal clinical trials. Pivotal clinical trials of recombinant human bone morphogenetic protein-2 are underway in patients with open tibial shaft fractures and in patients with a deficiency of the alveolar ridge. PMID- 11937872 TI - Carpal tunnel syndrome: relationship between clinical and patient-oriented assessment. AB - Clinical diagnosis of carpal tunnel syndrome usually is easy and sensitive, but frequently, it is based on history and referred symptoms when no motor or sensory deficits are observed during examination. In patients with carpal tunnel syndrome, a dissociation between the severity of the disease reported by the patient and the deficits clinically evaluated by the physician often are observed. The Italian Carpal Tunnel Syndrome Study Group did a multicenter study on 740 patients (1123 hands) with carpal tunnel syndrome to assess the aspects of dissociation between the patient's and the physician's quantification of the disease severity and the ability of the patients with carpal tunnel syndrome to do daily activities. Validated patient-oriented parameters assessing symptoms and hand functional status were used. The relationship between the physician's and the patient's measurements is strong with a linear significant correlation when analyzing the functional status of the hand but not so clear and simple when analyzing the symptoms. Patients with mild-to-moderate carpal tunnel syndrome seemed to function well, although severe symptoms may be reported by the patient; however, when nerve impairment becomes severe, the patient's hand function is extremely impaired although symptoms may be milder. The data show that the patient's point of view is reliable. Carpal tunnel syndrome seems to be an ideal model to evaluate the importance of patient-oriented measurements. PMID- 11937871 TI - Incidentally diagnosed Kienbock's disease. AB - The results of examinations of 14 patients with incidentally diagnosed Kienbock's disease were reviewed. These patients had not been examined previously, clinically or radiographically, and had not received any treatment for their condition. For 12 of these patients, Kienbock's disease was diagnosed incidentally on radiographic examinations obtained for other reasons, including carpal tunnel syndrome in four patients, Colles' fracture in three patients, pseudogout attack in the wrist in two patients, infection of the hand in one patient, osteoarthritis of the carpometacarpal joint of the thumb in one patient, and osteoarthritis of the metacarpophalangeal joint in one patient. The remaining two patients were diagnosed incidentally with Kienbock's disease based on radiographs obtained from a medical examination. Although radiographic findings revealed an advanced Kienbock's disease, current symptoms were mild in all patients, no problems with wrist pain were observed in activities of daily living or at work, and no treatment for Kienbock's disease was required. Appropriate treatment for Kienbock's disease should be considered carefully because some patients have no problems with activities of daily living or work for many years. PMID- 11937873 TI - Fluoroscopy-based multiplanar image guidance for insertion of sacroiliac screws. AB - A prospective controlled experimental study was done to assess the value of fluoroscopy-based, computer-assisted orthopaedic surgery for the insertion of sacroiliac screws and to compare this new technology with the conventional technique, using image intensification. In a simulated surgical setup, 140 cannulated screws were placed into the S1 and S2 vertebral bodies of 35 pelvic models. The screws were inserted under fluoroscopy-based image guidance or with the conventional technique. Different drills were tested with both techniques, including a 2.8-mm guide wire and a 5-mm solid drill. The 2.8-mm guide wire proved inaccurate with the computer-assisted image guidance system because of guide wire flexibility. Using the more rigid 5-mm drill, the results of computer assisted image guidance were comparable with the image intensifier technique. The radiation exposure during screw insertion was reduced considerably when using the image guidance system. System-specific requirements warranted adjustment of surgical technique and instruments. The major advantages of this new technology are immediate intraoperative image acquisition and provision of surgical guidance in as much as four planes simultaneously. The results of this experimental study are encouraging and have led to initiation of a clinical trial. PMID- 11937874 TI - Posttraumatic sagittal osseous bar in the spinal canal of an adult: a case report. AB - Diastematomyelia is a congenital anomaly wherein the distal spinal canal is bisected by a longitudinally oriented septum made up of fibrous tissue, cartilage, or bone. The main lesion is expressed by characteristic gait disturbances and dysfunction of the anal and vesical sphincters resulting from damage to the cord or cauda equina. Symptoms almost invariably begin during childhood. By the time symptoms appear, the damage is largely irreversible. In rare instances, the neuropathic expression of diastematomyelia is delayed until adult life. Only a dozen such cases have been reported. Described herein is the case of a patient in whom the typical symptoms appeared at 28 years of age. Decompression laminectomy and resection of the septum relieved the symptoms completely and permanently. The pathogenesis of nerve damage in diastematomyelia is thought to be different in patients with adult-onset disease than in children. This could explain why surgical treatment has a better prognosis in adults. PMID- 11937875 TI - Percutaneous approach to the fifth lumbar and first sacral disc. AB - Reaching the L5-S1 disc space through a posterolateral percutaneous approach can be challenging and, at times, disappointing when the iliac crest is too high and the angle between the posterior rim of the iliac bone and the lumbar vertebral column is too acute. The authors are introducing a technique through which the two necessary caudal and axial angles of approach are measured and used for precise passage of the initial trocar into the L5-S1 disc space. Specific bony landmarks and their topographic reflection on the lumbar skin are used in finding the caudal angle and calculating the axial angle. The technique has been used in 43 patients with persistently symptomatic L5-S1 disc protrusions, 27 of whom had high iliac rims. Access to the disc space was achieved in a timely manner in 41 patients who subsequently had percutaneous discectomy. The two unsuccessful procedures were because of the blockage of the access tunnel by an extremely swollen nerve root in one patient and a very large transverse process in the other patient. There were no other complications. Because of its accuracy, although the technique can be used routinely for all the posterolateral percutaneous approaches to the L5-S1 disc space, it specifically is useful for patients with high iliac crests. PMID- 11937876 TI - Adverse clinical events during cemented long-stem femoral arthroplasty. AB - The occurrence and risk factors for adverse clinical events associated with cemented long-stem femoral arthroplasty were studied. The hypothesis was that patients with femoral metastatic disease and previously uninstrumented canals were at higher risk for such adverse events. Fifty-five consecutive patients requiring long-stem femoral arthroplasty at two institutions were retrospectively reviewed. Adverse clinical events including hypotension, sympathomimetic administration, and O 2 desaturation were subclassified according to the timing of their occurrence. Adverse events occurred in 34 of 55 patients (62%), including coma in two patients and death in a third patient. The three catastrophic events occurred in patients with metastatic disease involving previously uninstrumented femoral canals. Desaturation was more frequent in patients with metastatic disease and previously uninstrumented canals compared with patients who had revision arthroplasty and patients with previously instrumented femoral canals. Preexisting medical illness was a significant risk factor in total adverse clinical events that included cement-associated adverse clinical events and cement-associated and postoperative hypotension. In long-stem cemented femoral components risk factors for adverse clinical events included metastatic disease, uninstrumented femoral canals, and preexisting medical conditions. These findings underscore the importance of appropriate patient selection, patient and family education, and anesthesia preparation before long stem cemented femoral arthroplasty. PMID- 11937877 TI - Preoperative health status of patients with four knee conditions treated with arthroscopy. AB - Normative preoperative Short Form-12 Health Survey components summary scores have not been reported for patients who have knee conditions treated with arthroscopy. During one calendar year, 269 consecutive patients with a knee injury (119 meniscus lesions, 73 anterior cruciate ligament ruptures, 54 articular cartilage lesions, and 23 with patellar instability) completed the Short Form-12 Health Survey before medical evaluation. Mental Component Scale and Physical Component Scale scores were computed. Scores were compared with previously published normative and age-specific data for several medical conditions and the United States general population. With the number of patients available, no significant differences were detected between knee conditions for the Mental Component Scale and Physical Component Scale scores. In general, Physical Component Scale scores among patients with knee conditions were similar to previously reported Physical Component Scale scores for patients with orthopaedic shoulder conditions and significantly lower than the United States population norms. These data can be used as historic control groups to represent patients with knee conditions requiring arthroscopy. Age-specific Mental Component Scale and Physical Component Scale scores are indicated when comparing groups with a limited age range. PMID- 11937878 TI - Comparison of biochemical markers of bone metabolism in serum and femur aspirates. AB - In 27 patients undergoing arthroscopy of the knee for treatment of meniscal diseases, biochemical markers of bone metabolism were measured in cancellous bone, and levels were compared with concentrations obtained from peripheral blood. Bone-specific alkaline phosphatase, osteocalcin, and collagen Type I metabolites (procollagen Type I N-terminal peptide and carboxy-terminal cross linked telopeptide) were studied simultaneously in serum and in the distal femur using a radioimmunoassay. Although levels of bone-specific alkaline phosphatase and osteocalcin did not differ between serum and cancellous bone, concentrations of collagen Type I metabolites were elevated significantly in healthy cancellous bone. The close correlations between bone and serum concentrations confirmed accuracy of results obtained from cancellous bone. The mean bone-to-serum ratio for alkaline phosphatase and osteocalcin was 1.1 and 1.2, respectively. Collagen Type I metabolite ratios of 2.2 (for carboxy-terminal cross-linked telopeptide) and 2.3 (for procollagen Type I N-terminal peptide) indicate that these markers are formed locally and then released into the circulation. Bone seems to be a major contributor of collagen Type I metabolites to the serum pool. PMID- 11937879 TI - Avoidance of medial cortical fracture in high tibial osteotomy: improved technique. AB - A new technique in oblique incomplete high tibial osteotomy that permits an increase of valgus correction while preventing fracture of the medial cortex was investigated. Closing wedge or opening wedge osteotomy was done on 23 tibias from cadavers before loading in an Instron testing machine. In seven specimens (Group 1), lateral oblique wedge osteotomy was done. In seven other specimens (Group 2), one medial oblique cut was made. In both groups, the osteotomy terminated 10 mm from the cortex and approximately 2 cm below the plateau. In nine specimens (Group 3), the osteotomy terminated in a 5-mm diameter hole, drilled in an anteroposterior direction, with its center positioned 10 mm from the medial cortex and 2 cm below the articular surface. The maximum angle of opening or closing before fracture of the cortex took place was recorded. In Groups 1 and 2, similar maximum correction angles were observed, 6.7 degrees versus 6.5 degrees, respectively. In Group 3, the stress relieving hole allowed the correction angle to be increased to 10 degrees. An oblique high tibial valgus closing wedge osteotomy with an apical drill hole allows a significant increase of the correction angle compared with the same osteotomy without a drill hole. Medial open wedge osteotomy offers no advantage over lateral closed wedge osteotomy in the maximum obtainable correction angle without failure of the cortex. PMID- 11937880 TI - Autologous chondrocyte transplantation for osteochondritis dissecans of the talus. AB - Eight patients with osteochondritis dissecans of the talus were reviewed. The average followup was 17.6 months (range, 8-26 months). The mean age of the patients was 31.8 years (range, 22-42 years). All patients had a preoperative examination, magnetic resonance imaging, and radiologic classification of the lesion through regular anteroposterior and lateral radiographs. Diagnostic arthroscopy and biopsies of healthy cartilage were done, which then had chondrocyte extraction and culture. After an average of 2.5 weeks, an arthrotomy, malleolar osteotomy, subchondral bone sclerosis debridement, and autologous chondrocyte transplantation were done. The postoperative treatment included nonweightbearing for 6 to 7 weeks, physiotherapy, and continuous passive motion. According to the postoperative evaluation score, all results were excellent to good with an average score of 0.6. No complications occurred. The arthroscopic reexamination of three patients at the sixth postoperative month and the radiologic evaluation of all patients showed the existence of cartilagelike tissue with complete coverage of the chondral defect. This method enables reconstructive intervention for unicompartmental defects of articular cartilage through implantation of new chondrocytes, establishment of a subchondral blood supply, and reconstruction of the articular surface. PMID- 11937881 TI - Two-stage revision for infected endoprostheses used in tumor surgery. AB - Thirty-four patients with infected massive endoprostheses had two-stage revision procedures to cure the infection and retain a useful functioning limb. An antibiotic impregnated cement spacer constructed with two cement gun liners and a Kuntscher nail provided temporary stability and also allowed a high dose of local antibiotic concentration. In three patients the infection never was controlled, and one of these patients had a successful second revision surgery. Six other patients had reinfection varying from 14 to 92 months after surgery. Risk factors for reinfection were previous radiotherapy and any additional operative intervention. The overall success rate for controlling infection was 91% at 1 year and 74% at 5 years. Six patients required amputation. The mean functional outcome for the retained limbs using the Musculoskeletal Tumor Society scoring system was 77%. Two-stage revisions seem almost as effective at controlling infection in massive endoprostheses as in conventional joint replacements, particularly if additional surgical interventions can be avoided. PMID- 11937882 TI - Resection replantation of the arm for sarcoma: an alternative to amputation. AB - Resection-replantation for primary malignant tumors of the arm as an alternative to forequarter amputation first was reported in 1995. Segmental resection of the upper arm and replantation of the distal segment to the proximal humerus as an alternative to shoulder disarticulation for a large tumor of the humerus is reported. This procedure provides the patient with an aesthetically acceptable single-segment arm, which is useful for activities of daily living and recreation. There has been no local recurrence or metastasis at 5 years followup. PMID- 11937883 TI - Incidence and mechanisms of infiltration of sarcomas in the shoulder. AB - The intraarticular infiltration of sarcomas around the shoulder was analyzed. Of 58 sarcomas located around the shoulder, which were treated between 1993 and 1998, 15 osteosarcomas, 12 chondrosarcomas, and one Ewing's sarcoma that abutted the shoulder were selected. Radiologic images of 28 tumors were analyzed and compared with pathologic examinations. In 17 of 28 sarcomas (15 in the proximal humerus and two in the scapula), joint infiltration was suspected radiologically because of the existence of a tumor mass probably inside the joint or disruption of the joint surface and protrusion of the tumor. Seven of 17 tumors with radiologically positive and zero of 11 tumors with radiologically negative findings showed histologically positive findings for tumor inside the joint. Four of 11 osteosarcomas and three of four chondrosarcomas of the proximal humerus showed histologic findings of joint infiltration. Pericapsular extension was seen in two patients, direct articular spread with a pathologic fracture was seen in two patients, direct articular spread was seen in one patient, direct articular spread with pericapsular extension was seen in one patient, and pericapsular extension with a pathologic fracture was seen in one patient. The current study showed that pericapsular or direct articular infiltration into the glenohumeral joint is a phenomenon in sarcoma around the shoulder. Exact assessment of tumor existence inside the joint can be difficult in osteosarcoma and chondrosarcoma, so abnormal radiologic findings should be regarded as proof of existence of tumor, and extraarticular wide resection should be planned. PMID- 11937884 TI - Aneurysmal bone cyst arising from a fibrous metaphyseal defect in a toddler. AB - This is a case of aneurysmal bone cyst arising from a fibrous metaphyseal defect of the proximal tibia in a toddler. A 32-month-old girl presented with physiologic bowing with a small metaphyseal defect at the medial aspect of the left proximal tibia. A large aneurysmal bone cyst, occupying almost the entire metaphysis of the proximal tibia, evolved from this lesion during the following 1 year 3 months. The lesion healed uneventfully after curettage and bone grafting. This case emphasizes the importance of regular followups to observe the clinical course of a fibrous metaphyseal defect during childhood. PMID- 11937885 TI - Intraarticular synovial sarcoma confirmed by SYT-SSX fusion transcript. AB - A case of intraarticular synovial sarcoma arising in the knee verified by detection of the tumor-specific SYT-SSX fusion transcript is described. Although it is extremely rare, synovial sarcoma may occur entirely within a joint space. The molecular assay for detecting a tumor-specific chimeric gene is a valuable tool for diagnosis of soft tissue sarcoma, especially for diagnostically difficult cases and tumors in unusual locations. PMID- 11937886 TI - Knee immobilization on meniscal healing after suture: an experimental study in sheep. AB - Immobilization and nonweightbearing may influence the healing of sutured meniscal lesions in the avascular zone. In 12 sheep, 3 to 4 months of age, a 0.5-cm longitudinal lesion was made in the middle segment and posterior portion of the medial meniscus of the left knee in the avascular zone. The lesion was sutured immediately. The knees of six sheep were immobilized with a monolateral external fixator, which crossed the knee, and the knees in the other six sheep were not immobilized. The animals were sacrificed 6 weeks after the operation. Specimens from the anterior meniscus were used for histologic and vascular studies, and the posterior meniscus was used for mechanical testing. Repair was observed in two of 12 sheep in the form of fibrous tissue. Fibrochondrocytes and synovial cells were involved. Meniscal fibrochondrocytes from the nearest meniscal tissue, synovial cells from the femoral and tibial meniscal surfaces, and fibroblastic cells migrating through the suture channel provide cell access to the meniscus lesion. Sutured menisci, immobilized or not, support less than 50% of a normal load and are mechanically weak. PMID- 11937887 TI - The effect of osteogenic protein-1 in an in vivo physeal injury model. AB - The physis has limited ability to undergo repair, and injury may result in growth arrest. Osteogenic protein-1 promotes bone formation in diaphyseal defects, chondrocyte proliferation, and matrix synthesis. The authors' goal was to determine if the presence of osteogenic protein-1 in a defect involving the physis would promote cartilage repair, and in doing so, to determine the effect of osteogenic protein-1 on physeal growth. An ovine model of growth plate damage was used, in which the proximal medial physis of the tibia was partially ablated. The defect was filled with a Type I collagen paste containing osteogenic protein 1 (350 microg) or collagen alone. Growth rate was measured at 4, 14, and 56 days, and the defects were analyzed histologically at 4, 14, and 56 days. Bone bridge formation occurred within the defect site. However, osteogenic protein-1 promoted outgrowth of the adjacent physeal cartilage. The physeal cartilage underwent expansion until the mineral forming within the defect site blocked its progress. The effect was localized because only that portion of the physis at the defect margin appeared to be affected. PMID- 11937888 TI - Posterior cruciate ligament rupture alters in vitro knee kinematics. AB - Isolated posterior cruciate ligament injuries usually are treated nonoperatively, although some patients remain symptomatic, and degenerative changes within the patellofemoral joint and the medial compartment of the tibiofemoral joint have been seen in followup studies. In vitro simulation of knee squatting was done to quantify the influence of the posterior cruciate ligament on tibiofemoral and patellofemoral kinematics. For five knee specimens, knee kinematics were measured before and after sectioning the posterior cruciate ligament, and compared using a Wilcoxon signed rank test. The only kinematic parameters that changed significantly after sectioning the posterior cruciate ligament were the tibial posterior translation and patellar flexion. The posterior translation of the tibia increased significantly between 25 degrees and 90 degrees flexion. The average increase in the posterior translation exceeded 10 mm at 90 degrees flexion. The patellar flexion increased significantly from 30 degrees to 90 degrees flexion. The average patellar flexion increase peaked at 4.4 degrees at 45 degrees flexion. Increased tibial translation could adversely influence joint stability. Increased patellar flexion could increase the patellofemoral joint pressure, especially at the inferior pole, leading to degenerative changes within the patellofemoral joint. PMID- 11937889 TI - Enhancement of pedicle screw fixation through washers. AB - This study examined the effect of washer usage on initial pedicle screw fixation and on the salvage of replaced pedicle screws, and the effect of minor adjustments of pedicle screws on insertional torque. Titanium, nontapered pedicle screws (6.5-mm in diameter and 35-mm in length) from one manufacturer and custom made 5-mm washers were used in the fixation of porcine lumbar spines. Insertional torque was measured with an electronic torque screwdriver and failure strength was determined by straight pullout of the screws using an MTS machine. Initial insertional torque values were significantly greater in pedicle screws placed with washers compared with screws placed without washers. When the screw placed without a washer was salvaged with the addition of a washer, a significant increase in insertional torque resulted. Pullout testing failed to show a significant difference between the screws that were placed with washers and the screws that were placed without washers. In the second part of the experiment, there was a significant decrease in insertional torque after backing out the screw as little as 90 degrees. This current study showed that (1) washers significantly increase the insertional torque of pedicle screws; (2) screws placed without a washer can be salvaged and replaced with a washer, which results in significantly increased insertional torque; and (3) backing out a pedicle screw 90 degrees significantly decreases its insertional torque. Washers can be used with pedicle screws to enhance the initial stability of the screw constructs, and to maximize insertional torque when screws need to be replaced, revised, or adjusted (backed out). PMID- 11937890 TI - Temperature rise during reamed tibial nailing. AB - The current study determined the temperature rise during reamed tibial intramedullary nailing in vivo. Eighteen adult patients were studied. The tibial medullary canal diameter ranged from 8 to 11 mm and was reamed to at least 1.5 mm above the required nail diameter with AO reamers. Reaming of the medullary cavity ranged from 9 to 12 mm before nail insertion. Intraoperative monitoring of the heat produced during reaming of the medullary cavity was done by inserting two platinum resistance thermometer probes into the cortical bone at the short isthmic segment of the tibial shaft. The probes were connected to a data logger, and temperature readings were taken every 5 seconds during each reaming procedure. The mean tibial temperature before initiation of reaming was 35.6 degrees C (standard deviation, +/- 0.5 degrees), and peak temperatures recorded were from 36.1 degrees C to 51.6 degrees C. A direct correlation was observed between temperature elevation and amount of reaming. With reaming above 10 mm, tibias with a canal diameter of 8 mm showed a statistically higher temperature rise compared with tibias with a canal diameter of 9, 10, or 11 mm. No patients had intraoperative or postoperative complications related to skin or bone thermal necrosis, and bony healing progressed uneventfully. The small amount of reaming required to insert a nail into a normal 9-, 10-, or 11- mm tibial canal does not seem to produce a clinical problem. Reaming smaller canals (8 mm) to a larger size may induce a significant heating effect. PMID- 11937891 TI - An 8-year-old boy with neck pain. PMID- 11937892 TI - External fixation of select intertrochanteric fractures with single hip screw. PMID- 11937893 TI - Section Editor's Commentary. Anti-TNF Therapies Have Eliminated the Need for Steroids in Pediatric Crohn's Disease. PMID- 11937894 TI - Section Editor's Commentary. We Still Are More Than Molecules. PMID- 11937895 TI - Section Editor's Commentary. Infliximab as First-Line Therapy for Crohn's Disease. PMID- 11937896 TI - Photophobia in anterior visual pathway disease. PMID- 11937897 TI - Photophobia as the presenting visual symptom of chiasmal compression. AB - Five patients with a chief visual complaint of photophobia were subsequently found to have compressive lesions of the optic chiasm. Visual acuity and visual field deficits were often subtle. Magnetic resonance imaging scanning revealed large suprasellar masses, including three pituitary adenomas, a craniopharyngioma, and a clivus chordoma. Photophobia resolved in all patients following treatment of the tumors. A compressive lesion of the optic chiasm should be considered in patients who experience persistent photophobia unexplained by ocular abnormalities. PMID- 11937898 TI - Clinical features of late-onset pseudotumor cerebri fulfilling the modified dandy criteria. AB - OBJECTIVES: To characterize the clinical features of patients with pseudotumor cerebri (PTC) fulfilling the Modified Dandy Criteria who were diagnosed at or after the age of 44 years. METHODS: We reviewed the medical records between 1987 and 1999 of 14 patients at a single institution who were diagnosed as having PTC at 44 years of age or older according to the Modified Dandy Criteria: neurologic manifestations attributable to generalized increased intracranial pressure, elevated cerebrospinal fluid pressure with normal cerebrospinal fluid composition demonstrated by lumbar puncture, and normal or small ventricles demonstrated by neuroimaging. We documented presenting symptoms and signs, significant medical conditions, and visual field follow-up. RESULTS: There were nine women and five men. Nine patients (64%) were obese. Five patients (36%) were asymptomatic. None presented with headache alone. Four patients (29%) had an identifiable cause of intracranial hypertension, including two with transverse sinus thrombosis, one with severe chronic obstructive pulmonary disease and cor pulmonale, and one with corticosteroid withdrawal after prolonged administration. During a median follow up of 2 years of 12 patients, visual fields remained stable in 8, improved in 3, and worsened in 1. CONCLUSIONS: In comparison with patients who have idiopathic intracranial hypertension, our small series of 14 patients diagnosed after the age of 44 years were more often men, were less often obese, were less symptomatic, and had identifiable causes of intracranial hypertension in a substantial minority (29%). The visual prognosis in this age group is generally good. Because the nonidiopathic causes of PTC would be overlooked by adhering to the Modified Dandy Criteria, we propose a modification that excludes patients who have dural venous sinus disease demonstrated on magnetic resonance imaging and those who may be exposed to medications or toxins or have systemic disorders that are known to increase intracranial pressure. PMID- 11937899 TI - Early resolution of third nerve palsy following endovascular treatment of a posterior communicating artery aneurysm. AB - A 69-year-old man underwent successful endovascular treatment of a posterior communicating artery aneurysm that had caused a third nerve palsy. Pupil size became normal within 10 days and ptosis and ocular ductions became normal within 3 weeks of the procedure. Based on the reported recovery rates of third nerve palsy after aneurysmal clipping, recovery may occur more rapidly in patients who undergo endovascular treatment. Further data are necessary to substantiate this hypothesis. PMID- 11937900 TI - Spasm of accommodation associated with closed head trauma. AB - Spasm of accommodation, creating pseudomyopia, is generally associated with miosis and excess convergence as part of spasm of the near reflex. It may also exist as an isolated entity, usually attributed to psychogenic causes. We present six cases of accommodative spasm associated with closed head injury. All patients were male, ranging in age between 16 and 37 years. The degree of pseudomyopia, defined as the difference between manifest and cycloplegic refraction, was 1.5 to 2 diopters. A 3-year trial of pharmacologically induced cycloplegia in one patient did not lead to reversal of the spasm when the cycloplegia was stopped. All patients required the manifest refraction to see clearly at distance. The pseudomyopia endured for at least 7 years following head trauma. This phenomenon may represent traumatic activation or disinhibition of putative brain stem accommodation centers in young individuals. PMID- 11937901 TI - Whipple disease with supranuclear ophthalmoplegia diagnosed by polymerase chain reaction of cerebrospinal fluid. AB - An elderly man developed acute progressive supranuclear ophthalmoplegia and other central nervous system manifestations that suggested Whipple disease. Results of small intestinal biopsy were negative but polymerase chain reaction testing of the cerebrospinal fluid confirmed the diagnosis. PMID- 11937902 TI - The torsional component of "horizontal" congenital nystagmus. AB - OBJECTIVES: To study the relationship between the major horizontal and minor torsional components of congenital nystagmus to elucidate the diagnostic importance, effects on vision, and pathogenetic implications of the torsional components. METHODS: We recorded the eye movements of 13 subjects with congenital nystagmus using a three-dimensional magnetic search coil technique over a 15-year period. The subjects fixated on stationary targets straight ahead and along the horizontal and vertical meridians. Six of the 10 subjects with horizontal congenital nystagmus were asymptomatic; the remaining 4 (plus two with a vertical component to their congenital nystagmus) had adult-onset symptoms. An additional subject without symptoms had a vertical congenital nystagmus component plus seesaw nystagmus; one of the symptomatic subjects also had seesaw nystagmus. RESULTS: In all 13 subjects, the horizontal and torsional cycles were phase locked, and positive horizontal (rightward), vertical (upward, if any), and torsional (clockwise) motion coincided in 10 subjects. That is, rightward horizontal eye rotation coincided with clockwise curvilinear motion (rightward and downward) of the upper pole of each eye. During the horizontal foveation periods, torsional motion was also of low velocity. In 2 of 13 subjects, the torsional waveforms differed from those in the horizontal plane; in others, the direction or the variation with gaze angle differed from that predicted by Listing. In each of the 13 subjects, the torsional components ranged from 8.16% to 94.42% (median, 32.94%) of the peak-to-peak magnitudes of the congenital nystagmus. In most cases, the measured torsion was far greater than that predicted by Listing's law for a worst-case analysis (range, 0.69-11.83%; median, 4.91%). The torsional components of the two subjects with seesaw nystagmus were 60.48% and 264.02%. CONCLUSIONS: The manner in which the horizontal and torsional components of "horizontal" congenital nystagmus were phase-locked made clinical detection of the torsional component difficult. Most "horizontal" congenital nystagmus is actually horizontal-torsional congenital nystagmus. Visual acuity during horizontal foveation periods is not significantly diminished by torsional motion. In only one subject did the torsional component of the congenital nystagmus have an amplitude equivalent to Listing torsion; in the other 12 subjects, torsion exceeded our estimate of what Listing's law would predict. The torsional components of the seesaw nystagmus in two subjects also greatly exceeded the torsion predicted by Listing torsion. The most parsimonious explanation for our data is that the cyclic torsion in congenital nystagmus was generated centrally and not a result of Listing torsion, mechanical crosstalk, or normal or abnormal extraocular-muscle (plant) dynamics. Further measurements are needed to confirm this hypothesis. PMID- 11937903 TI - Bilateral simultaneous central retinal artery occlusions in atherosclerosis. AB - An 82-year-old woman with atherosclerosis developed central retinal artery occlusions (CRAOs) in both eyes within an interval of 5 minutes. There was no evidence of a thromboembolic source, systemic hypotension, vasculitis, migraine, or hypercoagulable state. Virtually simultaneous binocular CRAOs have been rarely reported, and suggest the possibility of a systemic but still undefined "hypercoagulable trigger." PMID- 11937904 TI - Safety issues in magnetic resonance imaging. AB - Although generally considered safe, magnetic resonance imaging (MRI) has a number of safety issues, including the effects of high magnetic fields and radiofrequency pulses on the body, and on implanted devices, the side effects of contrast agents, toxicity during pregnancy, claustrophobia, and hearing loss. PMID- 11937905 TI - An interview with William F. Hoyt, MD. [interview by Lanning B. Kline]. PMID- 11937918 TI - Rapid visual recovery after coenzyme q10 treatment of leber hereditary optic neuropathy. PMID- 11937919 TI - Optic nerve enhancement in leber hereditary optic neuropathy: four years later. PMID- 11937920 TI - [Analysis of an internet site. Cubby]. PMID- 11937921 TI - [Bio-warfare, terrorism of tomorrow?]. PMID- 11937922 TI - [The role of the dermatologist in pediatric emergency]. PMID- 11937923 TI - [Borrelia strains that cause erythema migrans in Alsace, France]. AB - INTRODUCTION: In Europe, Borrelia burgdorferi sensu stricto, Borrelia afzelii and Borrelia garinii are known as pathogens among the Borrelia burgdorferi sensu lato group. Since it is not yet known which Borrelia are responsible for Lyme borreliosis in France, the objective of this study was to identify the species of Borrelia responsible for erythema migrans in the region of Alsace, France. PATIENTS AND METHODS: Eighteen patients with erythema migrans (EM) of more than 5 cm of diameter were included in this prospective study. All patients were investigated at the Strasbourg University Hospital. Patients were biopsied on the active border of their lesion. Cutaneous biopsies of the active border of the lesion were cultivated in BSK-H medium (Sigma) and analysed in vitro by PCR after 8 weeks of culture, using flagellin consensus sequences which are present in all species of Borrelia burgdorferi sensu lato group as primers. Species-specific oligotyping was used for species identification. RESULTS: Among the 18 patients biopsied, 7 had evidence of borrelia infection revealed by culture and/or PCR. Borrelia afzelii was detected in 4 patients and Borrelia garinii in three. CONCLUSION: These preliminary results appear to confirm that Borrelia afzelii and Borrelia garinii are the predominant borrelial species in EM lesions in our geographic area, as in other European countries. PMID- 11937924 TI - [Chronic eczematiform eruption in the elderly]. AB - INTRODUCTION: Eczematiform eruptions in the elderly represent a relatively frequent motive for consultation and may lead to repeated hospitalization. Their etiologic diagnosis is often difficult and explains the frequent relapses. The frequent relapses can be explained by the difficulty in determining their etiologic diagnosis. The aims of this study were: 1) to specify the evolving characteristics of these eruptions in elderly patients and 2) determine their etiology. PATIENTS AND METHODS: The inclusion criteria in this retrospective study were: patients aged over 65 at the time of diagnosis, presenting with extensive eczematiform eruption (> 20 p. 100 of body surface) and lasting for more than one month. Eczema on stasis dermatitis of the lower limbs and generalized contact eczema were excluded. Eighty-three patients followed between January 1990 and January 1999 were included. The clinical, biological, histological and evolving characteristics were analyzed. RESULTS: Mean age of patients was 77 +/- 8 years and the male female sex ratio was 2.4. Patients received a mean of 4.0 +/- 2.6 drugs/patient, consisting essentially of cardiovascular and psychotropic agents. The cutaneous eruption had evolved a mean of 12.5 months (1 to 48 months) before diagnosis. Eczema was pruriginous in 92 p. 100 of cases. Frequent relapses were observed in 68 p. 100 of cases. Precise etiologic diagnosis was retained in 48 patients (58 p. 100). This was disseminated contact eczema (n=19), lymphoma cutis (n=10), atopic eczema (n=7), scabies acariasis (n=6) and pemphigoid (n=6). No etiologic diagnosis was retained in the remaining 35 patients (42 p. 100). Comparison of the characteristics in the 2 groups showed excessive consummation of medicinal products (p=0.024), predominant eruption of sun-exposed areas (p=0.004) and a greater frequency of histological images of keratinocyte necrosis (p=0.0072) in patients presenting eczematiform eruptions of unknown etiology. DISCUSSION: These observations suggest the eventual responsibility of medicinal products in the occurrence of certain extensive and chronic eczematiform eruptions in the elderly. However, the delays of imputability of various causal drugs were often longer than those currently admitted for toxidermia, and the withdrawal of potentially imputable agents rarely led to spectacular improvement in the lesions. A case test report is in progress to specify this hypothesis. PMID- 11937925 TI - [Open trial of mycophenolate mofetil in the treatment of resistant pemphigus vulgaris]. AB - BACKGROUND: Pemphigus vulgaris is a severe autoimmune blistering disease of the skin and mucous membranes. In absence of treatment, mortality is high. In the past, prednisolone was the best treatment. Later, combination therapy with systemic prednisolone and other disease-modifying drugs was tried with better results. Unfortunately, some patients do not respond well to such treatment, or may exhibit multiple recurrences or complications. Some other patients may remain on high dose corticosteroids to maintain remission. OBJECTIVE: To evaluate the efficacy and safety of mycophenolate mofetil as a steroid sparing agent in the treatment of resistant pemphigus vulgaris. METHODS: We administered 2 g daily mycophenolate mofetil with systemic steroids to 10 patients with resistant and severe disease who did not respond to conventional therapy, or had multiple recurrences. RESULTS: Nine of the ten patients responded to treatment and showed complete clearance of lesions within 6 to 16 weeks of therapy. At the end of six months, the dose of prednisolone was significantly lower. Side effects were few and mild. After discontinuation of mycophenolate mofetil, 5 of the 9 patients relapsed. CONCLUSION: Mycophenolate mofetil is effective and safe as a disease modifying drug combined with prednisolone in the treatment of patients with resistant pemphigus vulgaris. To induce long lasting remission has to be administrated for more than 6 months. PMID- 11937926 TI - [Methi-resistant Staphylococcus aureus in dermatological practice: origin, risk factors and outcome]. AB - BACKGROUND: Increased frequency of methiresistant Staphylococcus aureus (MRSA) in inpatients is a day to day problem. OBJECTIVE: To determine the origin of MRSA, the causes for contamination, and potential complications in a department of Dermatology. PATIENTS AND METHODS: A retrospective study of patients hospitalized in a dermatology department with cutaneous MRSA during 1997-1998. We considered age, type and duration of dermatitis, geographic origin of patients on admission, previous hospitalizations, time between arrival in the ward and positive MRSA, and complications requiring systemic antibiotics. RESULTS: Out of 4579 of our patients, 53 (0.011 p. 100) had positive MRSA. They were on average 70.86 years old (26 to 97). The most common underlying dermatitis was leg ulcers (30) and foot sores (4). Dermatitis had lasted for more than 1 month in 48 patients. Most of the patients (40) had their MRSA on admission to our ward. Twenty six patients admitted from home had MRSA; only 9 had never been in an hospital. Six patients had diabetes mellitus. Thirty two patients healed with local treatment for their dermatitis. Thirteen patients have had some complications (erysipela 1, osteoarthritis 3, septicemia 2, febrile syndrome 9). DISCUSSION: Most patients with MRSA had leg ulcers or foot sores, confirming liability of chronic wounds to MRSA colonizing. Thirty one out of 40 patients with MRSA at arrival had been previously hospitalized. Hospitalization increases the risk of MRSA contamination. Treatment of MRSA is essentially that of the underlying dermatitis. However, it is still necessary to monitor the lesion for complications. PMID- 11937927 TI - [Prognosis of primary cutaneous lymphomas]. AB - INTRODUCTION: The assessment of prognosis is a major step in the management of primary cutaneous lymphomas, as it allows to give patients an accurate information and it directs the treatment choice. MATERIAL AND METHODS: We performed a literature review of global prognosis and prognostic factors in primary cutaneous lymphomas. We used survival as the main endpoint, in particular specific survival and relative survival which provide accurate estimates of lymphoma-related deaths. Independent prognostic factors identified by multivariate survival analyses were emphasized. RESULTS: Overall prognosis of mycosis fungoides has improved during the past decades, possibly because of an increased proportion of cases diagnosed at early stages. Five-year disease specific or relative survival rates of patients with T1 stage (patch/plaque disease<10 p. 100 of total skin surface), T2 (> 10 p. 100), T3 (tumor stage) and T4 (generalized erythroderma) are 100 p. 100, 67 to 96 p. 100, 51 to 80 p. 100 and 41 p. 100 respectively. Lymphomatoid papulosis and CD30+ primary cutaneous large T-cell lymphomas have an excellent prognosis, with 5-year survival rates of 100 p. 100 and 96 p. 100 respectively. CD30-negative primary cutaneous large T cell lymphomas have an aggressive clinical behavior (5-year disease-specific or relative survival: 15 to 21 p. 100). Among primary cutaneous B-cell lymphomas, immunocytomas and marginal-zone B-cell lymphomas are not life-threatening. Follicle center-cell lymphomas that arise on the head and trunk have also an indolent clinical course (5-year specific survival rates: 94 to 97 p. 100). However, few of these lymphomas are composed of more than 50 p. 100 of large cells with round nuclei (centroblasts and/or immunoblasts) and may have a more aggressive clinical course (5-year specific survival: 72 p. 100). Large B-cell lymphomas of the leg often occur in older patients and have a poorer prognosis (5 year specific survival rate: 52 p. 100). Cases with a single lesion and those with a predominance of large cleaved cells (large centrocytes) have a more favorable clinical course than those with multiple tumors or a round cell morphology. CONCLUSION: Clinical, histological and immunophenotypical prognostic factors which have been identified so far may reliably predict the survival outcome in primary cutaneous lymphomas. On this basis, therapeutic guidelines have been proposed. These prognostic data will have to be taken into account when evaluating new potential prognostic factors (e.g. immunophenotypic or molecular) and performing prospective clinical trials. PMID- 11937928 TI - [Delayed cerebral radionecrosis following radiation therapy of cutaneous squamous cell carcinomas of the head]. AB - INTRODUCTION: Three cases of cerebral radionecrosis occurring after radiation therapy for squamous cell carcinomas of the scalp are reported. This rare and poorly documented complication of radiotherapy is discussed. CASE REPORTS: Three patients presenting with squamous cell carcinomas of the scalp were treated with surgery and radiotherapy for recurrent or incomplete resection of squamous cell carcinomas of the head. X-ray doses range were 50 to 60 Grays in 22 to 24 fractions. Cerebral radionecrotic lesions were diagnosed 6 months to 14 years after irradiation, and were inconstantly associated with clinical symptoms. The patients were treated with systemic steroids, which were of limited efficacy in one of our patients. DISCUSSION: Little is known of cerebral radionecrosis following radiotherapy. This may be related to their rare occurrence and/or to the difficulties in establishing diagnosis. The delay of occurrence after radiotherapy can vary between a few months and several years, and the lesions are directly correlated with the doses and the fractionning of the X-rays. Intracerebral localization of the tumour is the main differential diagnosis. Localized and cystic forms of cerebral radionecrosis can be treated by surgery. Treatment otherwise relies on systemic steroids. PMID- 11937929 TI - [Intraepithelial capillary hemangioma?]. AB - INTRODUCTION: Histological examination of a biopsy of a lesion of the glans penis revealed blood capillaries within the malpighian epithelium. CASE REPORT: A 66 year-old man exhibited two circumscribed red plaques of the glans penis following posthectomy. Their histological examination revealed blood capillaries within the epithelium without basal membrane and pericytes between the endothelial cells and keratinocytes. Further evolution was towards lichen planus of the genital mucous membranes. In spite of a verrucous carcinoma which had been surgically removed, the evolution within a follow-up time of 6 years was favorable. DISCUSSION: Blood capillaries ascending in the epithelium of the genital mucous membranes have been observed in a special variant of Zoon's balanitis, the lichenoid, telangiectatic and purpuric balanitis described by Jonquieres in 1971. In our patient exhibiting a lichen planus of the glans penis, the occurrence of blood vessels in the malpighian layer of the epithelium may be interpreted as the consequence of an unusual epithelial and vascular regeneration of an erosive lichenoid lesion. It may also be interpreted as an original neoplasia, and could therefore be considered as a genital intraepithelial capillary hemangioma. PMID- 11937930 TI - [Rhabdomyolysis induced by pentamidine (Pentacarinat) during treatment of cutaneous leishmaniasis: 2 cases]. AB - INTRODUCTION: We report 2 patients who developed pentamidine-induced rhabdomyolysis during treatment of leishmaniasis. This potentially harmful side effect of pentamidine has only been reported exceptionally until now. CASE REPORTS: In January 2001, we treated 2 patients, aged 31 and 38 years, with 600 mg pentamidine administered intramuscularly twice in 48 hours. The two patients developed rhabdomyolysis with elevated CPK (respectively 30-70-x normal) and myoglobinemia (respectively 20-28-x normal) levels. Outcome was good under hyperhydratation and alkaline diuresis. DISCUSSION: This underestimated side effect of pentamidine treatment is not mentioned in recent articles on the topic. In case of such treatment however, these biological parameters should be monitored in order to establish the real frequency of this side-effect. PMID- 11937931 TI - [Cutaneous Waldenstrom's macroglobulinemia]. AB - BACKGROUND: We report the case of a patient in whom the first manifestation of Waldenstrom' s macroglobulinemia was specific skin lesions, treated with chlorambucil chemotherapy. CASE REPORT: A 76-years old woman was referred to us because of chronic red nodular lesions on her face. A biopsy specimen showed a dense lymphocytic dermal infiltrate and immunohistochemistry identified a monoclonal B lymphoid population with an IgM-kappa phenotype. The patient's disease was diagnosed as Waldenstrom's macroglobulinemia with cutaneous localization, on the basis of a high level of circulating macroglobulinemia and a lymphoplasmocytic infiltrate in the bone marrow expressing the same monoclonal IgM-kappa as in blood and skin. Treatment with radiotherapy (12 Grays) was unsuccessful. Chlorambucil (16 mg per day, 7 days per month) was then introduced with rapid disappearance of the skin lesions. Neutropenia led to withdrawal of this treatment after 4 courses. The skin lesions relapsed 18 months later and were cured with chlorambucil at a lower dose. DISCUSSION: Specific skin infiltrates have been rarely described during Waldenstrom's macroglobulinemia. Review of the literature showed eight cases of such lesions treated by chemotherapy with only two successes with oral cyclophosphamide and polychemotherapy (cyclophosphamide, vincristine and CCNU). Chlorambucil was used unsuccessfully three times. We hypothesize that primary resistance to alkylating agent and the small number of cases of cutaneous Waldenstrom's macroglobulinemia may explain the poor response to systemic chemotherapy previously reported. PMID- 11937932 TI - [Bullous pemphigoid induced by spironolactone]. AB - INTRODUCTION: The role of spironolactone as pemphigoid-inducing agent has recently been suggested. We report a new case of pemphigoid probably induced by spironolactone. The disease resolved after withdrawal of the diuretic drug containing aldactone. OBSERVATION: A 76 year-old patient was hospitalized for eczematiform lesions and severe pruritus. Histological and immunological investigations led to the diagnosis of pemphigoid. For several years, the patient had been treated with acarbose, amlodipine, fluvastatine, buflomedil, lysine acetylsalicylate and a spironolactone-furosemide association. On withdrawal of spironolactone alone, the cutaneous lesions regressed spontaneously within 15 days and no relapse was noted 30 months later. DISCUSSION: Numerous observations have suggested the role of certain drugs in the occurrence of pemphigoid. A recent test case study concerning drugs administered to patients suffering from pemphigoid has shown the significant association with spironolactone and neuroleptics. In our case report, the imputability of spironolactone is plausible. Because of the poor prognosis of pemphigoid, essentially due to iatrogenic complications, it is valuable to be able to delay the initiation of specific treatment for a few days, in patients receiving spironolactone. PMID- 11937933 TI - [Phenylbutazone-induced sialadenitis fever simulating angioedema]. AB - BACKGROUND: Drug-induced sialadenitis is uncommon and unrecognized. Drugs such as nitrofurantoin, nifedipine and methimazole have been reported to induce sialadenitis. However, phenylbutazone and oxyphenbutazone are the most frequently implicated agents. We describe a case of phenylbutazone-induced parotitis and submaxillitis with cutaneous and hepatic involvement. CASE REPORT: A 51 year-old woman who had received phenylbutazone for the past 6 days was hospitalized for diagnosis of Quincke's oedema. Clinical examination in fact revealed bilateral parotitis and submaxillitis. The patient had contracted mumps in infancy. Improvement was noticed 8 days after stopping the drug and treatment by glucocorticosteroid. Nevertheless a pruritic eruption with fever appeared. Laboratory data showed leukocytosis with neutrophilia, ESR of 75 mm/hr, hepatic cholestasis and cytolysis. Infectious and autoimmune causes were ruled out. The eruption spontaneously disappeared after 5 days. Laboratory studies 3 weeks later were normal. DISCUSSION: Quincke's edema diagnosis had been established too fast on "allergic past history" and patient interrogation. Complete clinical examination revealed the correct diagnosis of sialadenitis. This observation shows similarities with other publications: unbearable xerostomia appearing before sialadenitis and with a long course, parotitis with sub-maxillitis, 6 days delay after the first administration of phenylbutazone before fever, local evolution without complication, inflammatory biological syndrome with neutrophilia and absence of infectious cause. Pruritic maculo-papulous eruption and biological hepatic abnormalities are however rare. An hypersensibility mechanism is discussed. PMID- 11937934 TI - [Erythromelanosis follicularis faciei and colli]. AB - BACKGROUND: Erythromelanosis follicularis of the face and neck, originally described in Japan by Kitamura et al. in 1960, is characterized by a clinical set of three: well-demarcated erythema, hyperpigmentation and follicular papules. It affects the face and the neck generally on both sides. Since the original description, it has seldom been reported in the literature. CASE REPORTS: This paper reports two patients with unilateral presentation. DISCUSSION: Having discussed the various differential diagnoses all published cases were listed and analyzed. The prevalence of this disease appears higher than is shown by the limited number of cases reported in the literature. It deserves more recognition. Its nosologic and aetiologic frames still remain to be clarified. PMID- 11937935 TI - [Mastocytosis with symetric disposition in twin sisters]. PMID- 11937936 TI - [Seborrheic dermatitis in children]. PMID- 11937937 TI - [Contact allergy to formaldehyde]. PMID- 11937938 TI - [Dermatologist or dermato-venereologist?]. PMID- 11937939 TI - [Nocturnal pain in both ears]. PMID- 11937940 TI - [Erythematous-papular lesions]. PMID- 11937941 TI - [Purplish-erythematous plaque of the lower limb]. PMID- 11937942 TI - [Misleading figure (2)]. PMID- 11937943 TI - [Buccal pathology]. PMID- 11937944 TI - [Cutaneous manifestations of bioterrorism]. PMID- 11937945 TI - [Hereditary angioneurotic edema]. PMID- 11937946 TI - [Acne fulminans therapy]. PMID- 11937948 TI - [Isotretinoin (Roaccutane) and psychiatric disorders]. PMID- 11937949 TI - [Leon Lortat-Jacob (1873-1931)]. PMID- 11937950 TI - [Juvenile cellulitis in a 3-month-old Siberian Husky puppy]. PMID- 11937951 TI - ["Kokoti": a ritual on the way to extinction among the black half-casts of Guyana]. PMID- 11937952 TI - [Paediatric dermatology: a new life?]. PMID- 11937953 TI - [Bone evaluation in ten adults with cutaneous mastocytosis]. AB - BACKGROUND: To systematically evaluate the bone status, searching for osteoporosis in patients with cutaneous mastocytosis. PATIENTS AND METHODS: In a prospective study from March 1997 to June 2000, we included all new patients examined for cutaneous signs of mastocytosis. Past history, physical examination, skin biopsy, laboratory tests, bone densitometry, cytology and histopathology of bone marrow and other complementary investigations were performed in all these patients. RESULTS: Ten patients were included. Two patients had osteoporosis. Five others had osteopenia. Four patients had bone marrow involvement due to mastocytosis. One of these four patients had myelodysplasia. DISCUSSION: This study suggests better investigation of bone density and bone marrow in patients who have cutaneous mastocytosis. Systematic histopathology of bone marrow and osteodensitometry help to detect patients with systemic involvement. Bone densitometry is particularly effective for early detection of patients at risk for fracture and is of practical interest since patients with osteoporosis can now be treated with Pamidronate. PMID- 11937954 TI - [Acne in adult women: data from a national study on the relationship between type of acne and markers of clinical hyperandrogenism]. AB - INTRODUCTION: The role of hyperandrogenism in acne occurring or persisting in adult women is controversial. Studies reported have often been carried out in hospital settings. The aim of this nationwide prospective and descriptive study was to evaluate the frequency of clinical hyperandrogenism in a large number of adult acneic women visiting dermatologists in a non-hospital setting. PATIENTS AND METHODS: Three hundred and fifteen dermatologists completed clinical questionnaires concerning the next five female patients with acne at their private practices. These patients had to be between 25 and 45 years of age. The questionnaire covered patients' demographic characteristics, medical history, gynaecological status and acne history. Patients' acne, seborrhea and cycle disorders were described, as well as other signs suggesting hyperandrogenism, such as hirsutism and alopecia. RESULTS: A total of 1 135 questionnaires were analyzed. Nearly 50 p. 100 of the patients had major scalp or facial seborrhea, 18.4 p. 100 hirsutism, 7 p. 100 alopecia and 32.2 p. 100 menstrual cycle abnormalities. When these signs were present, acne was more often retentional, with more scarring and more widespread. CONCLUSION: This prospective study in a large number of patients in a non-hospital setting shows that acne in adult women is frequently associated with clinical hyperandrogenism. PMID- 11937955 TI - [Blue-red macules and pseudoatrophic macules in neurofibromatosis 1]. AB - INTRODUCTION: Blue-red macules are exception during neurofibromatosis 1. Between October 1988 and March 2000 in our cohort of patients, we systematically looked for these lesions. PATIENTS AND METHODS: Presence and number of blue-red macules were evaluated. Biopsies and photographs were proposed to patients. RESULTS: Forty-four patients out of 583 (39 y; range: 20-66 y) (7.5 p. 100; CI 95 p. 100: 5.5-9.8 p. 100) had blue-red macules mainly on the trunk. Histologically, blue red macules corresponded to neurofibromatous tissue infiltrating capillary blood vessels and venules. CONCLUSION: Blue-red macules are a peculiar type of neurofibroma and can therefore be considered as a criterion for the diagnosis of neurofibromatosis 1. PMID- 11937956 TI - [Blood pressure monitoring in infants with hemangiomas treated with corticosteroids]. AB - INTRODUCTION: Alarming hemangiomas can be successfully treated with corticosteroids. The risk of hypertension is poorly documented in this setting. We conducted a prospective study in infants with hemangiomas treated with orally administered prednisone. PATIENTS AND METHODS: Thirty-seven patients with rapidly growing complicated hemangiomas were enrolled between January 1998 and November 1999. Steroid dosages varied from 1 mg/kg/d to 5 mg/kg/d. Blood pressure measurements were performed first twice a month, then monthly at rest with a Critikon Dynamap device. Hypertension was defined as blood pressure superior to 110/60 mmHg, and borderline if superior to 110/60 mmHg on only one or two measurements. RESULTS: Increased blood pressure was found in seven infants (19 p. 100). One infant had hypertension requiring specific treatment and blood pressure was borderline on six patients at one or two occasions. Cardiac ultrasound examination was performed in five patients revealing two cases of myocardic hypertrophy, which regressed after stopping steroids and without relationship to hypertension. CONCLUSION: Even though the definition of hypertension in still arbitrary in infants, the measurement of blood pressure is necessary when steroid therapy is given for the treatment of hemangiomas. Blood pressure measurement can be difficult in very young children and overestimation is frequent if an inadequate device is used. The exact place of cardiac ultrasound remains to be defined in the management of alarming hemangiomas. PMID- 11937957 TI - [Dermatoscopy of pigmented lesions]. AB - Dermoscopy is a simple to use in vivo method for the early diagnosis of malignant melanoma and the differential diagnosis of pigmented skin lesions. It has been shown to increase diagnostic accuracy over clinical visual inspection in the hands of an experienced physician. This paper is a review of the principles of Dermoscopy as well as recent technological developments. PMID- 11937959 TI - [Cutaneous polyarteritis nodosa in children: three cases]. AB - INTRODUCTION: Polyarteritis nodosa is a necrotizing vasculitis of small and medium-size arteries. The cutaneous form of polyarteritis nodosa follows a chronic course, characterized by recurrent episodes limited to skin, muscles and joints. This entity differs from systemic polyarteritis nodosa in the absence of visceral involvement. This form is rare in children, we describe three cases. CASE REPORTS: We describe three girls with a mean age of 11 years (range: 8-13). They presented painful subcutaneous edematous nodules, arthralgia and fever. Physical examination revealed livedo reticularis (2 cases) and pharyngeal infection (1 case). Laboratory findings showed an inflammatory syndrome. Skin biopsy supported diagnosis of polyarteritis nodosa. The course was characterized by periods of remission disrupted by exacerbations, well controlled by salicylotherapy, colchicine, dapsone or penicillin. Corticosteroid therapy was used only for invalidating symptoms. There was no systemic involvement after 2, 5 and 6 years of follow up. DISCUSSION: Cutaneous polyarteritis nodosa in children must be suspected in presence of fever, subcutaneous nodules, livedo reticularis and arthralgia. Prognosis is usually benign, so we recommend no aggressive treatment. In view of the tendency to relapse, long-term follow-up is appropriate, before confirming diagnosis. PMID- 11937958 TI - [Celecoxib induced toxiderma with positive patch-test]. AB - INTRODUCTION: Celecoxib (Celebrex(R)) is a new generation non-steroidal anti inflammatory drug, recently introduced in France. We report a maculopapular rash due to this drug with positive patch-tests. CASE REPORT: A forty year-old man received with 3 tablets/day of celecoxib for intercostal pain. Nine days after initiation of treatment a maculopapular rash appeared. At the end of treatment, the eruption persisted for two days, then rapidly improved within one week. Six weeks later, patch-tests with celecoxib diluted at 20 p. 100 in petrolatum were positive at 48 hours. DISCUSSION: Cutaneous reactions due to celecoxib are rare. In our case report, the delay, clinical aspect, improvement on withdrawal of treatment and positive patch test all emphasize the imputability of celecoxib. We wish to underline the possible cutaneous reactions with this new non-steroidal anti-inflammatory drug. PMID- 11937960 TI - [Ofuji's papuloerythroderma: two cases treated with azathioprine]. AB - BACKGROUND: Ofuji's papuloerythroderma is a rare disorder, characterized by a generalized pruriginous eruption, sparing the folds. It predominates in the elderly. The pathology is still unknown but associations with lymphoma have been described. Various therapeutic approaches have been tried, most often including local and general corticosteroids and PUVA. OBSERVATION: Two patients aged 71 and 84 years presented red pruriginous macular rash sparing the abdominal folds. Eosinophilia and lymphopenia were observed. Cutaneous biopsies showed dermal lymphocytic and plasmocytic infiltrates with, in one case, eosinophil and neutrophil exocytosis. Clinical, biological and morphological investigations showed no association with other diseases such as cancer or lymphoma. Azathioprine permitted clinical and biological remission in both patients but had to be interrupted because of minor side effects (infection, gastroenterologic disorders) and corticosteroids were introduced in one case. DISCUSSION: We suggest that histological aspects, such as exocytosis, may represent a link between Ofuji's papuloerythroderma and lymphoma. Azathioprine led to clinical and biological improvement in our 2 patients. Because of its adverse effects, it could be proposed as second-line therapy in patients presenting resistance or intolerance to usual treatments. PMID- 11937961 TI - [Increased serum and urinary levels of silver during treatment with topical silver sulfadiazine]. AB - BACKGROUND: Argyria, induced by prolonged absorption, is often of professional or medical origin. We report two cases of per cutaneous intoxication with topical silver sulfadiazine. CASE REPORTS: A 64 year-old hypertensive, diabetic woman presented bilateral venous ulcers on the legs. She had applied 100 g of silver sulfadiazine 1 p. 100 cream per week for the past 18 months. Silver concentration in blood high: 38 microgram/l (N<0.5) and led to renal dysfunction, without ocular or hepatic abnormality. A 19 year-old woman was treated with topical silver sulfadiazine for thermic cutaneous burns on legs. Renal and hepatic function was normal but silver concentration in blood was high at 440 microgram/l (N<0) with urinary excretion of silver at 12 microgram/l (N=0). DISCUSSION: Silver, from prolonged and excessive use of topical silver sulfadiazine, deposits in large amounts throughout the body: skin, labial mucosa, gingiva, kidney, liver and cornea. Monitoring concentration of silver in blood and/or urine is necessary, especially in patients treated with silver sulfadiazine cream for cutaneous burns. Indeed, silver is rapidly absorbed through the burn wound. It provokes hepatic, renal and neurologic tissue toxicity. Renal and hepatic function tests are not correlated with serum silver levels. CONCLUSION: The potential for silver toxicity is a direct consequence of applying silver sulfadiazine to extensive burn wounds. Hence monitoring concentrations of silver in blood and/or urine of patients receiving this treatment is recommended. PMID- 11937962 TI - [Erythroderma induced by hypercalcitoninemia: two cases]. AB - INTRODUCTION: The most common cause of erythroderma is the extension of a previous cutaneous disease to the entire skin surface, as is the case in mycosis fungoides or drug reactions. Erythroderma related to endocrine disorders has only exceptionally been published. We report two cases of patients suffering from erythroderma probably induced by hypercalcitoninemia. OBSERVATIONS: Case 1. A 69 year-old man had a two year-history of fluctuating erythroderma, resistant to all topical or systemic therapy. After numerous unsuccessful investigations, a high serum calcitonin rate was detected. The pentagastrin test was highly positive, suggesting a medullar carcinoma of the thyroid. Thyroidectomy showed only hyperplasia of C cells. All skin signs disappeared within a few days after surgery. Two years later, there was no recurrence of the erythroderma and the serum calcitonin rate was normal. Case 2. A 58 year-old man suffered from congestive and pruriginous erythroderma for 2 1/2 months, which responded well to topical steroids. Biological investigations done for flushes, showed a raised calcitonin level and highly positive pentagastrin test. Thyroidectomy also revealed simple hyperplasia of the C cells. No recurrence of the erythroderma was noted during the 6 month follow-up. DISCUSSION: These are the first cases of erythroderma associated with hypercalcitoninemia. The causal role of calcitonin is probable in the first case, because of 1) long duration of skin signs before the hypercalcitoninemia; 2) quick post-operative recovery, in parallel with the decrease in calcitonin level; 3) absence of recurrence after thyroidectomy; 4) absence of other proven causes. Erythroderma may have resulted from dilation of the skin vessels, since calcitonin and calcitonin gene related peptide are powerful vasodilators. PMID- 11937963 TI - [Eccrine porocarcinoma with extensive cutaneous metastasis]. AB - BACKGROUND: Porocarcinoma is a malignant tumour of the eccrine sweat duct, arising from acrosyringium. The tumoral lesions involve the deep dermal tissue. CASE REPORT: We report the case of an 84 year-old woman, suffering from a porocarcinoma, extensively involving the major part of the left lower limb. The first symptoms appeared two years ago. Hundreds of metastatic papules and small nodules were present, isolated or confluent into large plaques. The clinical picture was very close to lymphangioma. Diagnosis was confirmed by histopathologic examination. Radiotherapy was useful short-term, allowing partial flattening of the lesions and improving lymphatic drainage, thus providing comfort for the patient. It did not prevent a later progression of the tumoral process. DISCUSSION: Porocarcinoma is a rare tumour that usually appears as a single nodule or a plaque, arising from a preexistent eccrine poroma, or developing de novo. Two histopathological variants are described: trabecular or epidermotropic. This latter form, observed in the present case, is more aggressive, leading to frequent local recurrences and/or metastases. Our report is exceptional: the literature shows only one other case with such widespread cutaneous involvement. The clinical course of our case is discussed. PMID- 11937964 TI - [Solitary circumscribed neuroma (palisaded encapsulated neuroma) of the oral mucosa]. AB - BACKGROUND: The solitary circumscribed neuroma or "Palisaded Encapsulated Neuroma" is an infrequent benign tumor of the skin of adults, often localised on the face, around the orifices. The involvement of the oral mucosa or of the vermilion border of the lips is rare. CASE REPORTS: In our 5 cases, a small painless and more or less protruding nodule had been present for years on the mucosa of the hard palate (50 year-old man, 43 year-old woman), the soft palate (63 year-old woman), the vermilion border of the lower lip (48 year-old woman), and the gingiva (26 year-old man). The non specific clinical aspect did not permit a correct diagnosis. DISCUSSION: Histologically, the solitary circumscribed neuroma is formed by a proliferation of Schwann cells arranged in interlacing bundles, featuring one or several well-limited nodules, sometimes surrounded by a thin capsule. At first sight, this aspect might sometimes mimic schwannoma or neurofibroma. The lesion is interspersed with a variable amount of axons. An association with neurofibromatosis or MEN syndrome type IIb has never been found. Even after incomplete excision the solitary circumscribed neuroma does not recur, a fact supporting a reactive rather than a neoplastic origin. PMID- 11937965 TI - [Laser and lupus]. PMID- 11937967 TI - [Stewart-Treves' syndrome: long term survival]. PMID- 11937966 TI - [Acute generalized exanthematous pustulosis due to meladinine with positive patch tests]. PMID- 11937968 TI - [Asymptomatic tumefaction of the ear]. PMID- 11937969 TI - [Hyperpigmented painful cutaneous nodule of the thigh]. PMID- 11937971 TI - [Post zoster pain]. PMID- 11937972 TI - [Syphilitic alopecia]. PMID- 11937970 TI - [Photosensitivity in children]. PMID- 11937973 TI - [Recurrent Bloch-Lapiere's circinate erythema-like psoriasis]. PMID- 11937975 TI - [Urinary excretion of 6-sulphatoxymelatonin in normal subjects: statistical approach to the influence of age and sex]. AB - A radioimmunoassay of urinary 6-sulphatoxymelatonin (a-MT6s) was performed in 90 normal subjects: 44 males and 46 females (17-67 years). Patients treated with betablokers or antidepressants were not included in this study. Urine samples were collected over three periods of time: 7 to 11 p.m., 11 p.m. to 7 a.m., and 7 to 11 a.m. Between 11 p.m. and 7 a.m., the subjects slept in their normal environment and had not ingested alcohol for 24 hours. We searched for a possible relation between urinary a-MT6s excretion (expressed in ng/l/h) and age. From 7 to 11 p.m. and from 7 to 11 a.m. no significant relation could be found. On the contrary, between 11 p.m. and 7 a.m. there was a significant relation indicating decrease of a-MT6s secretion with increasing age. Several linear or non-linear curve patters were tested: Boltzmann sigmoid (1(st), 2(nd), and 3(rd) degree), polynomial curves. The Boltzmann sigmoid showed the best fit judging by the r squared value (0.152) and the runs test (p=0.64). On this curve the inflection point was located at 53 4 years (SDM, standard deviation of the mean). From 19 to 45 years, the upper sigmoid plateau was located at 1381 91 ng/l/h (SDM). The decrease was found between 45 and 60 years and the lower sigmoid plateau then stabilized at 467 370 ng/l/h (?SDM). In the study group, there was no significant difference between men and women according to the Mann-Withney test. Finally, use of oral contraceptives did not affect urinary a-MT6s (Mann-Withney). PMID- 11937976 TI - [Hungry bone syndrome after surgical treatment of severe primary hyperparathyroidism: about 3 cases]. AB - Persistant hypocalcemia occurring after surgical treatment of severe primary hyperparathyroidism may be due to transient or permanent hypoparathyroidism but also to a bone disease. We report three cases of hypocalcemia after surgery of large parathyroid adenoma or hyperplasia in women. Plasma calcium, phosphate and PTH levels are in accordance with Hungry Bone Syndrome (HBS). HBS is related to both excessive bone demineralization and turn over. It is a major importance to distinguish HBS from surgical hypoparathyroidism in order to start early the appropriate treatment given for a long period. PMID- 11937977 TI - [Pituitary germinoma presenting as a pseudotumoral lymphocytic hypophysitis in a man]. AB - A 45-year-old man presented with headaches and extraocular muscle palsy due to a sellar mass extending into the right cavernous sinus. Hormonal determinations revealed a gonadotrophic insufficiency. A transsphenoidal surgical removal revealed a lymphocytic hypophysitis with fibrosis and necrosis. Rapid growth of the pseudotumor was noted despite a high dose steroid therapy (1 mg/kg/d) for a month. Further biological and histopathological investigations were performed. They showed a high cerebrospinal fluid (CSF) B-human chorionic gonadotropin (ss HCG) level of 12 UI/L (normal<5 UI/L), normal plasma BHCG level, and undetectable CSF and plasma alpha-fetoprotein levels. The tumors cells showed a positive reactivity for placental alkaline phosphatase and for vimentin. These findings were consistent with an inflammatory lymphocytic process caused by an intrasellar germinoma. Chemotherapy was ill-tolerated and external radiotherapy was ineffective. PMID- 11937978 TI - [Diagnostic strategy in persistent hyperparathyroidism. Report of two cases and review of the literature]. AB - Ectopic adenoma is the main cause of surgery failure in the treatment of primary hyperparathyroidism (HPT). Localization of the abnormal glands is therefore necessary before a reoperation. We report two cases illustrating the interest of MRI and selective venous sampling of parathormone. In two patients with persistent hyperparathyroidism, ultrasound and sestamibi scanning failed to localize the ectopic glands. MRI suggested the presence of an adenoma in the mediastinum for the first patient and inside the thyroid for the second. In both cases, the localisation was confirmed by venous sampling and the reoperation was a success. We review the recent literature on this topic and we propose a diagnostic approach of persistent HPT. PMID- 11937979 TI - [Multihormonal bilateral petrosal sinus sampling in Cushing's disease: radiological, surgical and pathological correlations]. AB - Multihormonal bilateral petrosal sinus sampling (BPSS) has been proposed to improve corticotroph microadenomas prediction of lateralisation. Few series have simultaneously compared data of pituitary MRI, surgical findings and multihormonal BPSS. Seven patients (6F/1M) with Cushing's disease, mean age at diagnosis of 35 years (range 24-55) were prospectively studied to compare radiological and multihormonal BPSS data with surgical and pathological findings. In untreated patients, simultaneous measures of ACTH, TSH and prolactine (PRL) were done at time 0, 7, 15, 22 minutes after CRH (500 microgram) and TRH (200 microgram) stimulation. An intersinus gradient of 1.4 was considered as a lateralisation. All microadenomas were identified during surgery, diameters ranged from 2 to 7 mm. All patients were in long-term surgical remission. Pathological studies confirmed a tumoral tissue with ACTH immunostaining in 6/7 cases and PRL in 3/7 cases. Pituitary MRI correctly identified tumors in 4 cases, the remaining tumors were not seen. Basal and stimulated intersinus gradients of ACTH, TSH and PRL were homolateral in 6/7 cases and were coincident with surgical findings in 4/7 cases. The other three cases were contralateral to MRI and surgical data. In conclusion, simultaneous gradient of ACTH, PRL and TSH did not improve lateralisation prediction in this series. Hormonal hypersecretion was homolateral in six cases whereas pathological studies demonstrated a mixed secretion in only three cases. A preferential pituitary draining could explain these discordances. Data from our series and from others (done with CRH stimulation and ACTH-PRL measures) strongly suggest a paracrine interaction between tumoral and normal pituitary tissue. PMID- 11937980 TI - [Polycystic ovary syndrome: treatment with insulin-sensitizing agents]. AB - Polycystic ovary syndrome (PCOS) is a common disorder. Its prevalence is 5 to 10% in women of reproductive age. PCOS is associated with hyperinsulinism and insulin resistance. The pathophysiological situations has lead many authors to study the action of insulin-sensitizing agents on menses, ovulation rate, and pregnancy in patients with PCOS. Metformin (a member of the biguanide family), is used for treatment of type II diabetes mellitus in obese patients. Although metformin restores cyclic pituitary- gonadal function and improves fertility, it can decrease levels of androgen and LH and increase levels of SHBG in women with PCOS. Trooglitazone (a member of the thiazolidinedione family) has been withdrawn from use because of its liver toxicity. Troglitazone improves ovulation and hisrsutism in women with PCOS without change in body mass index. Other similar drugs with less liver toxicity may be useful for the treatment of PCOS. D-chiro inositol is a mediator of insulin action and improves ovulatory cycles. Most of the studies reported have not been randomized but the results appear to be quite promising. These drugs may provide a substantial advance in the treatment of women with polycystic ovary syndrome. PMID- 11937981 TI - [Risks and challenges for medical school training]. PMID- 11937982 TI - [Ewing's sarcoma]. PMID- 11937983 TI - [Treatment of Ewing's sarcoma with 2 different protocols]. AB - Since 1985, 54 with localized Ewing's sarcoma of bone were treated at the Onco Orthopedics Clinic of the Sofia University Hospital (Sofia, Bulgaria). Thirty-two patients received the classical four-drug combination with doxorubicin, dactinomycin, vincristine, and cyclophosphamid. The last 22 patients were treated with six-drug protocols where cisplatinium and ifosfamid were added to the standard four-drug regimens. Local treatment was adapted to the age of the patient and the location and volume of the tumor. Fourteen patients received radiotherapy alone, 24 postoperative radiotherapy and 16 surgery alone as local treatment. At a mean follow-up of 6 years, the disease free survival rate reached 44%. Eleven patients had a good local response and 25 developed metastatic disease. Statistical analysis confirmed the deleterious effect of pelvic localizations and large tumor volume on prognosis and demonstrated the dramatic improvement obtained with the six-drug protocol (5-year disease free survival 61% versus 23%) and surgery (61% versus 25%). PMID- 11937984 TI - [Panniculitis revealing alpha-1 antitrypsin deficiency. Report of 3 cases]. AB - We report panniculitis revealing alpha-1 antitrypsin deficiency in 3 patients with different Pi phenotypes. The first patient, a 16-year-old woman, had inflammatory skin lesions on the abdomen for 6 months. The lesions regressed spontaneously. Serum alph-1 antitrypsin level was normal but the Pi phenotype was MS. The second case was observed in a 56-year-old man who developed erythematous subcutaneous nodules on the abdomen, legs and buttocks in a trauma context. Serum alpha-1 antitrypsin was very low and the Pi phenotype was ZZ. The third patient was a 40-year-old woman who presented red swelling nodules on the legs. Her serum alpha-1 antitrypsin level was at the lower limit of normal and the Pi phenotype was MZ. Alpha-1 antitrypsin deficiency is an autosomic codominant inherited disorder characterized by inefficient or non-functional serum alpha-1 antitrypsin. The principal clinical manifestations are panlobular emphysema and cirrhoses. About 30 cases of panniculitis have been reported in the literature. In patients presenting panniculitis, we suggest studying the Pi phenotype to determine functional deficiency even if the serum level of alpha-1 antitrypsin is normal. PMID- 11937985 TI - [Referral cycle of the rheumatoid patient]. PMID- 11937986 TI - [Team management of rheumatoid arthritis]. AB - The main objectives of team management of rheumatoid arthritis are to stop structural damage of joints and to reduce functional, psychological, socioprofessional and economic consequences. Team management requires the collaboration, around the patient, of a rheumatologist, a nurse, a psychologist, a physiotherapist, an occupational therapist, an orthopaedic surgeon at the same time, in the same place. More and more patients wish to manage their disease by themselves. Team care should not be proposed to every patient; it must be reserved to patients whose condition required such an approach because of the severity of the disease, comorbidity, psychological or socioprofessionnal difficulties. Team management should be personalized. Utility of team management is now accepted; out-patient administration is as effective as in-patient one. A good educational program is very important. However, search is still needed to define optimal modalities of team management and tools to measure the efficiency of this approach. PMID- 11937987 TI - [Aspects of the radiological evaluation of rheumatoid arthritis]. AB - This paper gives an overview of several aspects of the radiographic evaluation of rheumatoid arthritis (RA). Radiographs give important information about the structural damage caused by RA. On a group level, the natural progression follows a rather linear cause, but on an individual patient level, this can vary widely. Feet are often affected earlier and to a larger extent than hands. Both hands and feet give a good overall impression of the damage in the large joints and are sufficient to monitor the patient. It is recommended to take annual films of hands and feet to monitor patients. Several scoring methods are available for application in clinical trials. Most widely used are the Sharp and Larsen methods with their modifications. For use in clinical practice the SENS method is more feasible. Several drugs are capable of retarding radiographic progression. However, it is difficult to compare results across trials and these difficulties are elaborated. PMID- 11937988 TI - [Factors predictive of severity in rheumatoid arthritis]. AB - Rheumatoid arthritis (RA) is a very heterogeneous disease with multiple forms. Some are very mild with few or no overt lesions or joint deformations, others are very severe leading to rapid joint destruction and major functional handicap. The best therapeutic strategy for patients with early stage RA cannot be uniform, but must be adapted to each individual patient, depending on disease severity. This fact is becoming even more important with the development of new treatments which have had a major impact on therapeutic strategies. No prognostic factor has reached universal acceptance. There are however an increasing number of arguments allowing better recognition of prognosis during the early stages of the disease. High ESR, CRP, positive rheumatoid factor with a high IgM (and IgA) titer, the presence of one of the HLA DRB1*04 alleles associated with RA, and early development of radiological lesions are all factors of prognosis. PMID- 11937989 TI - [Perspectives for drug treatment in Alzheimer's disease]. AB - The last decade was very fruitful in neuropharmacology and notably in the therapeutic strategies of dementia and Alzheimer's disease (AD). The amount of data, information and breakthroughs is nevertheless difficult to apply in direct relationship with patients. The present review aims at classifying information according to their origins: epidemiology, clinical trials, neurosciences. A guide for drug prescription in Alzheimer's disease is thus warranted and becomes clearer, sure that, in the next future modifications and new strategies will appear. The main goal of the present review is to summarize the state-of-the-art for a non specialist in AD. PMID- 11937990 TI - [Inhalation therapy]. PMID- 11937991 TI - [Special issue of bone pathology] PMID- 11937993 TI - [Flat adenomas: should we reconsider colonic endoscopy and colorectal cancer prevention?]. PMID- 11937995 TI - [Primary thyroid lymphomas: clinicopathologic study of 30 cases and review of the literature]. AB - During a both retrospective and prospective study of thyroid cancers treated in the Basse Normandie between 1960 and 1999, we have identified 32 patients with thyroid lymphoma. The correct diagnosis was made initially in 69% of all cases. In the other cases, the diagnosis was secondarily corrected after review of the pathological material. According to the REAL classification, 7 (21%) corresponded to low grade MALT lymphomas, 2 to low grade lymphomas, 10 to high grade MALT Lymphomas and 10 (31%) to high grade lymphomas, one plasmocytoma and two unclassified lymphomas. According to the Ann Arbor classification, stage was IE for 56%, IIE for 19%, IIIE for 3% and 9% for IV. Median survival was 28 months with a mean at 61 months. 20 patients died (62%), 12 from the lymphoma and 8 from intercurrent causes. The overall survival at 5 years was 36% (9 5% CI 16 54%). A comparison of our results with those of the literature was performed. PMID- 11937996 TI - [Flat neoplasia of the colon and rectum: definitions and illustrations]. AB - Thanks to recent progresses in colonoscopy non-polypoid lesions, even very small, can be detected and removed per endoscopy. They are defined endoscopically as flat polyps or flat lesions and correspond microscopically to neoplasia in more than 30% cases. This review presents the definitions and different types of flat adenomas and flat early adenocarcinomas as established by Japanese authors. The use of appropriate definitions and classifications and the need of a perfect method in preparing the samples before microscopic examination are underlined. Flat adenocarcinomas tend to invade the submucosa at a smaller size than carcinomas arising in polypoid adenomas. They result either of the malignant transformation of flat adenomas or of a de novo type histogenesis. Flat early adenocarcinomas invading the submucosa and measuring less than 1 cm belong to the latter category. PMID- 11937997 TI - [Does malignant fibrous histiocytoma exist?]. AB - Malignant fibrous histiocytoma (MFH) has come to be regarded as the most common malignant neoplasm of the mesenchymal soft tissues. It designates a spectrum of tumors which share morphologic features that allow their inclusion in a distinct clinicopathologic setting, although being not uniform in their histogenesis and pathogenesis. Clinicopathologic variants include the following: the storiform pleomorphic form of MFH, the myxoid type of MFH, the giant cell type of MFH and the inflammatory type. The latter group, the angiomatoid variant, has been reclassified within the fibrohistiocytic tumors of low malignant potential. Tissue culture, ultrastructural and immunohistochemical studies have both endorsed or refuted the validity of the concept. As a whole, these morphologic studies which attempted to characterize MFH were not able to delineate specific markers or to describe the phenotype of this sarcoma of supposed fibrohistiocytic lineage. There is growing evidence that MFH is a second component in another sarcoma and represents a morphologic modulation resulting from tumor progression. Recent cytogenetic and molecular genetic investigations are consistent with that hypothesis: a comparative analysis between the most frequent genomic imbalances observed in series of MFH and leiomyosarcomas (LMS) demonstrated that both tumors had similar recurrent imbalances. Immunohistochemical and molecular biologic investigations have shown similar targets of chromosome deletions in both tumors. A new classification of soft tissue sarcoma based on molecular parameters is nevertheless premature. The morphologic characterization of MFH and its sub-types provides the clinician with unique information in the management of these tumors, by identifying a spectrum of tumors with well-recognized clinical profiles. PMID- 11937999 TI - [Arrector pili myositis in a patient with dermatomyositis: an unusual histological feature]. AB - Dermatomyositis is a rare inflammatory disease with a difficult diagnosis on cutaneous biopsies, since alterations are often little marked and non specific. A 52-year-old patient had severe paraneoplastic dermatomyositis. He had an erythematous rash on the trunk, and erythematous papules on the back of the hands. The first histological cutaneous lesion was a striking arrector pili myositis. Arrector pili myositis has rarely been described, and is thought to be linked with Wong type eruption occurring in dermatomyositis. Arrector pili myositis might constitute an helpful sign in the diagnosis of dermatomyositis, but its specificity and sensitivity are unknown. We compared this case to four previously reported cases. In these five cases, arrector pili myositis was not associated with a particular sub-type of dermatomyositis or with poor prognosis. PMID- 11937998 TI - [Langerhans cell histiocytois of the thyroid: a rare disease not to be ignored]. AB - Thyroid involvement by Langerhans cell histiocytosis is rare. We report the case of a ten-year old boy who presented with a 5 cm goitre. He was treated for diabetes insipidus 14 months before. Thyroid isotopic scan showed hypoactivity of right lobe and revealed a cold left nodule. The patient was treated by right lobo isthmectomy with adjuvant corticotherapy and chemotherapy (vinblastine). He is well with 12 months follow-up. Microscopic analysis demonstrated a diffuse infiltrate of thyroid parenchyma by sheets of CD1a positive Langerhans cells associated with lymphocyte foci. This case is remarkable by the abundance of Langerhans cells and scarcity of eosinophils. The diagnosis of thyroid langerhans cell histiocytosis should not be ignored in both children and adult patients. PMID- 11938001 TI - [Acute herpetic hepatitis: clinico-pathological study of two cases]. AB - Two cases of acute herpetic hepatitis affecting immunocompetent adults after surgery, are reported. They point out the relevance of liver biopsy for the diagnosis of these generally unrecognized infections. Liver biopsy allows not only diagnosis but also the identification of the virus on tissue sections with immunohistochemistry or in situ hybridization. The prognosis of herpetic hepatitis which can be correlated with the extension of apoptosis, remains poor despite the appearance of antiviral drugs. An apoptosis score could represent an interesting prognostic factor whose value remains to be assessed. PMID- 11938000 TI - [Sarcomatoid carcinoma of small intestine: diagnostic problems raised by an unusual tumor]. AB - Sarcomatoid carcinomas involving the intestinal tract are rare and usually associated with poor prognosis. We report a case of sarcomatoid carcinoma of the ileum, diagnosed in a 61-year-old man. The patient presented with acute intestinal occlusion. Surgical resection of the ileum was performed. At macroscopic examination, two large polypoid masses were found. Frozen section examination suggested the diagnosis of malignant stromal tumor. At histological examination, both tumors were formed by pleiomorphic, large spindle cells, presenting numerous mitoses and marked nuclear atypia. Immunohistochemical examination showed that tumor cells coexpressed vimentin and epithelial markers (cytokeratins, EMA). The final diagnosis was monomorphic sarcomatoid carcinoma. The patient deceased with metastatic disease after 3 months of follow-up. This report underlines the potential diagnostic problems raised by this unusual type of carcinoma and emphasizes the role of immunohistochemistry in assessing the correct diagnosis. PMID- 11938002 TI - [Epididymal tumor in a six-months-old infant]. AB - A rare tumor of the epididymis was discovered in a 6-month-old infant. Macroscopically, the tumor had a black focal color. Immunohistochemistry staining and electron microscopy led to the diagnosis of mealnotic neuroectodermal tumor, or progonoma. Prognosis of progonoma, a benign tumor, is generally good, but malignant transformation has been reported. PMID- 11938004 TI - [A polyposis which is not out of breath!]. PMID- 11938003 TI - [Role of cytology in the diagnosis of Rhodococcus equi infection]. AB - A 59-year-old HIV-positive man developed necrotizing right-lobe pneumonia with abscess formation. Bronchial aspirates and bronchoalveolar lavage showed numerous inflammatory cells, mainly histiocytes and neutrophils, and intracellular and extracellular Gram- and Ziehl-Nielsen-positive cocci and coccobacilli. Rhodococcus equi was identified. The patient died one week after diagnosis and necropsy confirmed the presence of R. equi nectrotizing pneumonia. PMID- 11938005 TI - [A tumour not so clear]. PMID- 11938006 TI - [An unusual cause of genital bleeding]. PMID- 11938007 TI - [An unusual occipital nodule]. PMID- 11938008 TI - [Tissue microarrays, or the advent of chips in pathology]. AB - Tissue microarray technology is a new method used to analyze several hundred tumor samples on a single slide allowing high throughput analysis of genes and proteins on a large cohort. The method consists to core tissues from paraffin embedded tissue donor blocks and placing them into a single paraffin block. Despite the low amount of tissue analyzed by tissue microarray, different studies have demonstrated a high concordance of protein expression between this technique and the conventional tissue sections. PMID- 11938009 TI - [Protocol for examination of orchidectomy specimen: proposal for a standardized reporting form]. AB - We present practical guidelines for the examination of orchidectomy specimens and propose a standardized worksheet for the pathology report of germinal tumors. PMID- 11938022 TI - Diabetes in Africans. Part 2: Ketosis-prone atypical diabetes mellitus. AB - Diabetes is increasing with ageing and changes in lifestyle in populations of African ancestry as described in the first part of this review. Apart from classical type 1 and Type 2 diabetes, atypical presentations are observed in these populations, especially "tropical" and "ketosis-prone" atypical diabetes. Ketosis-prone atypical diabetes that has been classified by ADA as idiopathic Type 1 diabetes or Type 1b is the most common atypical form. It is characterised by an acute initial presentation with severe hyperglycaemia and ketosis, as classical Type 1 diabetes. In the subsequent clinical course after initiation of insulin therapy, prolonged remission is often possible with cessation of insulin therapy and maintenance of appropriate metabolic control. Metabolic studies showed a markedly blunted insulin secretory response to glucose, partially reversible with the improvement of blood glucose control. Variable levels of insulin resistance are observed, especially in obese patients. Pancreatic B-cell autoimmunity is an exceptional finding. Association with type 1 susceptibility HLA alleles is variable. The molecular mechanisms underlining the insulin secretory dysfunction are still to be understood and may involve gluco lipotoxicity processes, glucagon dysregulation, effect of stress, or may be genetically determined. The present review summarises the available clinical and metabolic features and suggests some pathogenetic hypotheses and principles of management for the ketosis-prone atypical diabetes of the Africans. PMID- 11938023 TI - Sulfonylurea receptor -1 (SUR1): genetic and metabolic evidences for a role in the susceptibility to type 2 diabetes mellitus. AB - The pancreatic B-cell ATP-sensitive potassium channel (K(ATP)) is composed of two distinct subunits, an inwardly rectifying ion channel forming the pore (Kir6.2), and a regulatory subunit, namely the sulfonylurea receptor-1 (SUR1), which binds this widely used class of insulin-secreting drugs. Mutations in the genes encoding Kir6.2 and SUR1 may result in familial persistent hyperinsulinemic hypoglycaemia of infancy, demonstrating their role in the regulation of insulin secretion. Studies in various populations with different ethnic background provided evidence that various alleles of single nucleotide polymorphisms (SNPs) in the SUR1 gene, and to a less extent in the Kir6.2 gene, confer a significantly increased risk for the development of type 2 diabetes mellitus (T2DM). Allelic variations of these SNPs were shown to modulate insulin secretion and insulin sensitivity in vivo, thus providing a pathophysiological background to explain their contribution to the genetic susceptibility to T2DM. The aim of this review is to summarise and discuss the significant results of recent literature on the implication of K(ATP), and particularly of SUR1, in the genetic and pathopysiological mechanisms of T2DM. PMID- 11938024 TI - Effect of omega-3 fatty acids on lipid peroxidation and antioxidant enzyme status in type 2 diabetic patients. AB - BACKGROUND: This study was conducted to investigate the effect of omega-3 fatty acids on lipid peroxidation and antioxidant enzyme activities in non-insulin dependent diabetic patients. METHODS: Thirty-four non-insulin dependent diabetic patients were selected for this study and they were initially treated with antidiabetic drugs alone for one month. This was followed by supplementation with omega-3 fatty acids (1,080 mg of EPA and 720 mg of DHA per day) along with the antidiabetic drugs for a period of two months. RESULTS: No change in glycaemic control was observed in diabetic patients at the end of two months of omega-3 fatty acids therapy along with antidiabetic drugs. The combined treatment significantly reduced serum triglycerides (2.07 +/- 0.94 mmol/l, before combined therapy vs 1.54 +/- 0.49 mmol/l after combined therapy, P<0.05) and increased HDL cholesterol levels (0.93 +/- 0.099 mmol/l, before combined therapy vs 1.04 +/- 0.098 mmol/l after therapy, P<0.01). The raised lipid peroxide levels (5.14 +/- 0.61 micromol MDA/l in controls vs 6.36 +/- 1.56 micromol MDA/l in diabetic patients, P<0.001) were significantly decreased in these patients after the combined therapy (6.36 +/- 1.56 micromol MDA/l, before combined therapy vs 5.16 +/- 0.7 micromol MDA/l, after combined therapy, P<0.01). Among the erythrocyte antioxidant enzymes, the Glutathione peroxidase activity was increased (32.5 +/- 9.9 U/g Hb/min, before combined therapy vs 42.25 +/- 4.6 U/g Hb/min, after combined therapy, P<0.01) while no change was observed in Catalase (99.7 +/- 30.4 KU/g Hb before combined therapy vs 85.35 +/- 23.41 KU/g Hb, after combined therapy) and Superoxide dismutase activities (2.6 +/- 1.04 U/mg Hb/min, before therapy vs 3.01 +/- 1.08 U/mg Hb/min, after combined therapy) after the 2 months of combined treatment with antidiabetic agents and omega-3 fatty acids. CONCLUSION: Supplementation with omega-3 fatty acids has beneficial effects on serum triglycerides, HDL-cholesterol, lipid peroxidation and antioxidant enzymes, which may lead to decreased rate of occurrence of vascular complications in diabetes. PMID- 11938025 TI - Relationship between polymorphisms in the renin-angiotensin system and nephropathy in type 2 diabetic patients. AB - BACKGROUND: Renin Angiotensin system is involved in renal function and its polymorphisms may influence diabetic nephropathy. ID ACE polymorphism modulates ACE level whereas M235T AGT polymorphism is involved in arterial hypertension. The A1166C AT1R polymorphism is involved in arterial hypertension and in diabetic retinopathy. METHODS: Two hundred thirty five type 2 diabetic patients were enrolled in this transversal study. Data were documented for clinical characteristics of the population, HbA(1c), urinary albumin excretion, presence of retinopathy or antihypertensive treatment. Polymorphisms were analyzed by PCR techniques. The patients were divided into 3 groups: group 1, without nephropathy (n=118), group 2, microalbuminuria (n=78), group 3, macroalbuminuria (n=39). RESULTS: Diabetes duration was longer (p<0.001), retinopathy (p<0.001) and antihypertensive treatment (p<0.02) were more frequent in group 3 compared to group 1 and 2. The I/D ACE and M235T AGT polymorphisms were not differently distributed between the three groups. In contrast, the CC genotype of the AT1R polymorphism was overrepresented in group 2 (p=0.021). The presence of the CC AT1R genotype considerably increased the incidence of albuminuria after 10 years of diabetes (AA vs CC p=0.01), particurlarly in men. No effect was seen with I/D ACE and M235T AGT polymorphisms. CONCLUSION: In conclusion, we observed an interaction of A1166C AT1R polymorphism with diabetes in men but not of I/D ACE and M235T AGT polymorphisms. PMID- 11938026 TI - Clinical expression and insulin sensitivity in type 2 diabetic patients with heterozygous mutations for haemochromatosis. AB - BACKGROUND: Elevated iron metabolism indices as well as liver enzymes abnormalities have been reported in type 2 diabetic patients. The aim of this study was to determine the clinical and biological characteristics of overweight or obese type 2 diabetic subjects, with and without heterozygosity for HFE gene mutation (C282Y or H63D). We also assessed their insulin sensitivity and B cell function. METHODS: 90 patients (age and diabetes duration: 61 +/- 11 and 12 +/- 8 years [mean +/- 1 SD]) were included. BMI was 32 +/- 6 kg/m(2). HbA(1c) was 8.9 +/- 1.8%. HFE genotyping was performed by PCR and restriction enzyme cleavage. Insulin sensitivity and B cell function were measured by the Homeostasis Model Assessment (HOMA). RESULTS: Heterozygosity for C282Y (wt/C282Y) or H63D (wt/H63D) allele was found in 11 and 12 subjects respectively. There were no major differences in clinical status and iron parameters according to the single allelic presence of C282Y or H63D. However, systolic blood pressure [BP] was lower when such mutation was present. Insulin sensitivity and B cell function (HOMA) were comparable. When the cohort was divided according to gender, we found higher serum iron in females with than in those without HFE mutation (91 +/- 27 vs 73 +/- 25 microgram/dl;P=0.049), while a transferrin saturation index above 45% was observed in 36% of females with a mutation (vs 7% in wt/wt;P=0.06). When analysis was performed according to the presence of each particular mutation, we observed a transferrin saturation index higher than 45% in 60% of wt/C282Y patients vs 21% in the wt/wt group (P=0.008). A significantly lower BP was also identified in wt/C282Y patients. Cholesterol-HDL was 38 +/- 11 vs 46 +/- 12 mg/dl in wt/C282Y and wt/wt subjects, respectively (P=0.045). There were no differences in iron status, BP or lipids between wt/wt and wt/H63D subjects. CONCLUSION: Type 2 diabetic patients, in particular females, with mono-allelic C282Y mutation, had slightly increased iron parameters. Systolic BP and cholesterol-HDL were also lower in wt/C282Y subjects. No difference in insulin sensitivity or B cell function was observed in the presence of mono-allelic HFE mutations. PMID- 11938027 TI - Prevalence of the missense mutation Gly574Ser in the hepatocyte nuclear factor 1alpha in Africans with diabetes. AB - BACKGROUND: Clinical presentation and natural history of diabetes are somewhat different in Black Africans compared to Caucasians. This peculiar disease course could be at least partly related to a specific genetic profile that has not been studied in this population. METHODS: Medical backgrounds, anthropometric and biologic parameters were obtained from 69 diabetic subjects in Dakar, Senegal, in 1998. Blood anti GAD and Islet Cell Antibodies were studied, using RIA and immunofluorescence assay. The HNF-1alpha gene was sequenced searching the Gly574Ser mutation, previously described in MODY 3. RESULTS: Among these 69 diabetic patients, 11 (16%) were found to have the G574S mutation affecting the HNF-1alpha. These 11 patients carrying the mutation were compared respectively with the 58 non carriers. Mean age (57.5 yr. +/- 11 vs 51.1 yr. +/- 15) and duration of diabetes (11.9 vs 6.7 yr), were similar in the two groups. BMI was not different in patients with the mutation (26.3 vs 23.3, p=0.06). Metabolic control (Glycosylated hemoglobin) was poor in the two groups (9.5% vs 9.2%). Chronic complications were equally found in the patients, but no mutation carrier had macroangiopathy. None of the anti GAD positive or ICA positive patients had the mutation. CONCLUSIONS: The HNF-1alpha Gly574Ser mutation was found in 16% of cases in a 69 diabetic patients group in Senegal. Diabetes was as severe as in non carriers of mutation. This mutation has been implicated in atypical diabetes of Afro-American children. The study confirms its prevalence in Africans with diabetes. PMID- 11938029 TI - Measurement of accidental urinary insulin loss from a dislocated intraperitoneal insulin catheter. AB - We report the case of a type-2 diabetic woman who received continuous intraperitoneal insulin infusion and developed deterioration of metabolic control by accidental insulin loss into urine (54 U per day) as a consequence of catheter migration which probably resulted in bladder wall injury. Due to iodine allergy of this patient, an analyte addition procedure for insulin quantification in urine had to be applied to allow proof of insulin loss from the catheter tip before as well as reversal to zero insulin excretion after implantation of a new intraperitoneal port and a shorter catheter. The lost fraction of insulin accounted nearly completely for the difference between pre- and postoperatively required insulin doses (146 versus 88 U per day). PMID- 11938028 TI - Effect in vitro of cyclic nucleotides-elevating agents on nitric oxide production by human granulocytes from type 2-diabetic patients. AB - BACKGROUND: The present study was designed to investigate the hypothesis that cells from ill patients and from healthy subjects may have different reactivities under metabolic stimulation. METHODS: The study was performed with granulocytes from non-diabetic subjects and from type II -Non Insulin Dependent Diabetes mellitus (NIDDM) patients. The nitric oxide (NO) generation was comparatively determined by the nitrite concentration (micromolar of nitrite) after cell incubation in the presence of cyclic nucleotide-elevating agents. RESULTS: Our results showed an inverse reactivity for granulocytes from diabetic patients when compared to non-diabetic subjects. Granulocytes were incubated in the presence of drugs that elevate the intracellular level of cyclic AMP aminophylline (AMF), dibutyryl cyclic AMP (dbcAMP)], cyclic GMP [8.Br. cyclic GMP(8.Br.cGMP) or levamisole (LEV)]. The cyclic AMP-elevating agents (AMF and d bcAMP) inhibited NO production by granulocytes from non-diabetic subjects and activated cells from diabetic patients. By contrast, cyclic GMP-elevating agents (8.Br.cGMP and LEV) activated cells from non-diabetic subjects and inhibited granulocytes from diabetic patients. The activation of NO generation by cyclic nucleotides was blocked by pretreatment of granulocytes with L-NAME. CONCLUSION: The authors describe for the first time that both cyclic AMP and cyclic GMP were able to modulate nitric oxide production in human granulocytes and that cell reactivity in ill patients (diabetic) showed altered and inverse response in comparison to granulocytes from healthy subjects. This inverse reactivity possibly reflects a disease-induced adapted metabolic response. The consequences of this altered metabolic response on host defense and inflammation may be speculated, but further experiments are needed to confirm this hypothesis. PMID- 11938030 TI - Spontaneous physical activity in children: a disturbing factor? PMID- 11938032 TI - [The use of telephone for counseling diabetic patients: a descriptive and pedagogic approach]. AB - This descriptive study of telephone interviews, conducted between the staff of a diabetes unit and patients, is intended to clarify a professional manner in which the telephone should be used as part of patient care. Having examined simulated calls from diabetic patients, analysis of the interviews conducted by the doctors, nurses and members of the office staff in a hospital unit demonstrates wide differences in the content, the quality of the content, and the interview strategy. An interpretation of the results signals the importance of the manner in which the staff member answers, a knowledge of patients' genuine needs, and an ability to adequately acknowledge the emotional state of the patient. Structuring telephone conversations, allowing sufficient time for patient responses, avoids over-long interviews. The decisions taken by the staff member answering the call, in terms of advice and suggestions of actions to take, must be systematically backed-up by ensuring that the patient has properly understood what has been said. The study poses a number of questions, notably concerning the specific requirements demanded by these new remote-care services in the organization of care within a hospital unit, and the professional approach to the use of the telephone as part of the relationship with patients. Recommendations are given concerning the general principles governing a telephone interview and the definition of training objectives for healthcare professionals. PMID- 11938031 TI - [Factors associated with change in diabetes knowledge from childhood to adolescence]. AB - BACKGROUND: A multicentric cohort of 142 children with insulin-dependent diabetes has been longitudinally studied to evaluate if their diabetes knowledge was modified by time and to assess the factors associated to this change. METHODS: A knowledge scale, previously translated and validated in French (TDK), was independently completed by the children and their parents at inclusion (T0) and 4 years later (T4). RESULTS: Mean age of the children was 10.2 years at T0. Mean knowledge score of the parents did not differ between T0 and T4 but mean score of the children was significantly higher at T4 than at T0 (22.2 +/- 5.7 vs 26.9 +/- 3.8, p<0.001). Age-adjusted knowledge score of children at T4 was significantly correlated to compliance to treatment (r=0.23, p<0.01). The factors associated with knowledge score of the children at T4 were: age at T4 (r=0.49, p<0.001), knowledge score of children at T0 (r=0.59, p<0.001), school results of the children (r=0.18, p=0.04), educational level of the mother (r=0.21, p=0.01), family income (r=0.19, p=0.03), knowledge score of parents at T0 (r=0.16, p=0.09), number of diabetes summer camp periods (r=0.19, p=0.03). Multivariate analysis showed that these 7 factors accounted for 59% of the variance in predicting knowledge score of the children at T4 (p<0.001). After adjustment, age at T4, knowledge score of the children at T0 and educational level of the mother stayed significantly associated with the knowledge score at T4. CONCLUSION: These results emphasise the importance of diabetes education programs specifically developed for children with the aim of improving diabetes knowledge at adolescence. PMID- 11938034 TI - [Role of NOD2 gene in Crohn's disease]. PMID- 11938033 TI - [Is HbA1c appropriate for the screening of diabetes in general pratice?]. AB - The measurement of glycated haemoglobin (HbA(1c)) is a practical and more sensitive tool than fasting plasma glucose (FPG) in screening type 2 diabetes in current practice. Its use has been limited so far by the variability of the analytical methods. The standardization process is going on, and many laboratories are currently using valid methods. Our study is consistent with the results of other groups who recommended this measurement to identify undiagnosed diabetic patients, that are about 25% to 30% in the French population. The demonstration was provided through a survey including a screening step by both HbA(1c) and G0, and a second exam with a 2 hr OGTT in a sample of positive screenees according to at least one criterion (HbA(1c) >=6% or G0 >=1.26 g/L), as well as in a sample of negative screenees. We showed that nine confirmed diabetic subjects out of ten had HbA(1c) >=6% at the screening step, while only a half had G0 >=1.26 g/L. Conversely, 22% of the positive screenees according to HbA(1c) were not confirmed as diabetic by the OGTT, including however more than half with abnormal glucose values. A chart for practical use is proposed to define patients at risk, the process of screening, and the patient follow-up according to the results of the tests. PMID- 11938035 TI - [Mycophenolate mofetil in refractory Crohn's disease after failure of treatments by azathioprine or methotrexate]. AB - Failure or intolerance of treatments by azathioprine (AZA) or methotrexate (MTX) is an important problem in the maintenance treatment of refractory CD. Results of the alternative use of mycophenolate mofetil (MMF) are conflicting. AIM: To assess the efficacy and tolerance of MMF in patients with refractory CD included in a multicentre and retrospective study. PATIENTS AND METHODS: Twenty patients (5 men, 15 women, median age 33 years) have been enrolled. The disease was located in the small intestine (n=7), colon (n=5) or in both ileon and colon (n=8). Five patients had perineal disease. Neighteen patients had an active CD and 13 of them received steroids (30 mg daily). All of them had been treated by AZA (failure, n=5; intolerance, n=15) and/or MTX (failure, n=12; intolerance, n=6). The dose of MMF was 750 mg to 2 g daily for a median of 4 months (range: 4 days-21 months). Response to MMF was determined according to an Harvey-Bradshaw index<4 and the possibility to taper steroids. RESULTS: Four patients (AZA: failure, n=2; intolerance, n=2; MTX: failure, n=2; intolerance, n=1) achieved remission under MMF. Three of them had an active CD before start of MMF, 1 patient was already in remission. Those 4 patients remained in remission during follow up (10-18 months). One of the 5 patients with perineal involvement achieved a complete closure of perineal fistula. Ten patients had to stop the medication for a median of 4 months (range: 2-12 months) because of inefficiency. Eleven patients had early adverse reactions and 5 of them had to stop the medication within 4 days to 3 months: pancreatitis (n=2), toxidermia (n=2), diarrhea and abdominal pain (n=1). CONCLUSION: In this short cohort of 20 patients with refractory CD, either intolerant or non responder to AZA or MTX, treatment with MMF resulted in only 20% of success. Intolerance to MMF was observed in 25% of patients. PMID- 11938037 TI - [Inappropriateness of hospital days and causes of failure in a Gastroenterology and Internal Medicine ward]. AB - OBJECTIVES: To analyse patients' features linked to hospital inappropriateness and to highlight causes of inappropriate days in a Gastroenterology and Internal Medicine ward of a teaching hospital. METHODS: Appropriateness of patients' hospital days (2 months activity) was assessed using the French version of criteria of the Appropriateness Evaluation Protocol. Reasons of inappropriate hospital days were identified through a questionnaire based on patients' need. RESULTS: Two hundred and twenty patients were studied. Among the 2151 hospital days assessed, 880 (41%) were inappropriate. Two different groups of inappropriate stays were brought up. In the first group, the inappropriate period duration was short (<=5 days) and patients were not different from those of the appropriate group. In the second group, the inappropriate period duration was long ( > 5 days) and 710 days (33%) were inappropriate. Patients were elderly, lived alone and their disease did not concern the gastrointestinal tract. During inappropriate days, they expected access to less technical facilities than the short stay medical ward. CONCLUSION: The socio-demographic and medical features of the patients from the long duration inappropriateness group should help to limit inappropriate hospital days: a significant economic and organizational stake for patients, hospital and public interest. PMID- 11938036 TI - Serum measurements of pancreatitis associated protein in active Crohn's disease with ileal location. AB - BACKGROUND AND AIMS: Pancreatitis-associated protein (PAP) is a pancreatic stress protein also expressed in the ileum but not in the colon. Its serum concentration is increased in patients with small bowel inflammation due to untreated celiac disease. We searched to determine whether PAP could be a serum marker for ileal location of active Crohn's disease (CD). METHODS: A multicenter prospective study was conducted, including 54 healthy controls and 124 patients with CD of whom 38 had quiescent ileal or ileocolonic disease (group A), 45 had active ileal or ileocolonic disease (group B), 18 had quiescent colon-only CD (group C), and 28 had active colonic disease (group D). Active disease was defined by a Crohn's disease activity index > 150 and serum C-reactive protein (CRP) > 10 mg/mL. Location of lesions was assessed by endoscopy. PAP was assayed in serum, the upper threshold for normal values being 50 ng/mL. RESULTS: In group B, 27 patients (60%) had elevated serum PAP, compared to one in group A (2.5%), one in group C (5.3%), three in group D (10.7%) and none in the control group (P<0.01). By contrast, serum levels of C-reactive protein did not differ between patients with active CD and either ileal location (group B) or pure colonic location (group D) (38 +/-10.5 vs 41.6 +/- 6.4 mg/mL, NS). Within group B, serum PAP concentration was correlated with none of the epidemiological, clinical or biological data available. Increased serum level of PAP diagnosed ileal location in active CD with a sensitivity of 60%, a specificity of 94%, a positive predictive value of 84% and a negative predictive value of 81%. CONCLUSION: Elevated serum PAP (> 50 ng/mL ) is significantly associated with disease activity and ileal location PMID- 11938038 TI - [5-fluorouracil: metabolism and current indications in digestive cancer treatment]. PMID- 11938039 TI - [Clinical research is no longer for sale!]. PMID- 11938040 TI - [Risks and precautions for the use of lamivudine in patients with chronic hepatitis B virus infection]. PMID- 11938041 TI - [Registry of liver biopsies from hepatitis C infected patients in the Alpes Maritimes (France). Results from the first 2 years]. AB - OBJECTIVE: To perform a descriptive analysis of patients with chronic hepatitis C based on a local registry of liver biopsies. PATIENTS AND METHOD: Collection of clinical, biological and histological data from all HCV-infected patients who underwent liver biopsy between January 1997 and December 1998 in the Alpes Maritimes (France). RESULTS: One thousand and fifty six patients including 924 who lived in the Alpes-Maritimes (515 male, 409 female, mean age: 44.9 years old) were included. Intravenous drug use (30.1%) was the major suspected source of infection before blood transfusion (28.2%). Among intravenous drug users, 38% of patients were infected with genotype 1a and 37.4% with genotype 3. The METAVIR fibrosis severity score was distributed as follows: F0: 10.8%, F1: 53.7%, F2: 15.9%, F3: 14.7%, and F4: 4.9%. In a multivariate analysis adjusted for the duration of infection, independent risk factors associated with the severity of fibrosis were age at contamination >=30 years, genotype other than 1a and alcohol intake >=50 g/day. Determination of HCV antibody and liver biopsy were performed an average of 12.5 and 14 years after presumed date of contamination, respectively. CONCLUSIONS: These data provide a clearer view of the impact of this condition in this area and could help to define a comprehensive policy for patient management. PMID- 11938043 TI - [Revision of the December 20, 1988 law. Common platform for propositions from learned societies, organizations and patient associations]. PMID- 11938042 TI - Lamivudine therapy of chronic hepatitis B in three groups of patients: non transplanted patients, liver recipients, and kidney recipients. AB - OBJECTIVE: As conflicting results have been observed by some authors in liver recipients, the aim of the study was to evaluate lamivudine therapy in 3 groups of patients with chronic hepatitis B: non-transplanted patients, liver and kidney recipients. METHODS: All patients were studied for clinical symptoms, hepatic enzymes, hepatitis B virus (HBV) serology, serum HBV DNA load, and HBV polymerase genotype (mutations associated with lamivudine resistance). RESULTS: During the 48-144 week-long follow-up (mean: 75 weeks), 23 non-transplanted patients, 5 liver and 6 kidney recipients were studied. A sustained biochemical and virological response was obtained in 19 out of the 23 non-transplanted patients and in 4 of 6 kidney recipients, while the 5 liver recipients did not respond. After the development of lamivudine resistance, mutations rtM204V and rtL180M were detected in all studied patients, mutation rtM207I in one, with similar results from traditional nucleotide sequencing and a commercial line probe assay. CONCLUSION: The poor response to lamivudine in liver recipients requires further studies. PMID- 11938045 TI - [Synopsis: Diagnosis and treatment of hepatocellular carcinoma in patients with cirrhosis]. PMID- 11938046 TI - [Comments on case report: A 2.5 cm liver nodule discovered at ultrasound in a patient with controlled hepatitis C]. PMID- 11938047 TI - [Diagnosis and treatment of hepatocellular carcinoma. Questions for Professor Michel Beaugrand]. PMID- 11938049 TI - [Acute pancreatitis and pancreatic reactions due to Salmonella. A study of 6 cases]. AB - Numerous infectious agents may induce acute pancreatitis. We report 6 cases of Salmonella-associated acute pancreatitis. Pathogenesis of this type of pancreatitis is related to multiple factors that may coexist in the same patient. The typical clinical picture consists in epigastric pain, nausea, vomiting, diarrhoea and fever. Imaging techniques often disclose minimal changes such as moderate swelling of a part or of the totality of pancreas. As outcome is usually good, conservative treatment and antibiotics may be sufficient to ensure recovery. PMID- 11938051 TI - [Ductal adenocarcinoma of the pancreas associated with anthracosilicosis]. PMID- 11938050 TI - [Giant hemangioma of the liver revealed by an inflammatory syndrome]. AB - We report two cases of giant hemangioma of the liver revealed by a clinico biological syndrome including fever, right upper quadrant pain and a biological inflammatory syndrome, whereas liver function tests and blood cell count were normal. This clinical presentation may help in the diagnosis of giant hemangioma of the liver. PMID- 11938052 TI - [Retractile mesenteritis mimicking acute appendicitis]. PMID- 11938053 TI - [Atrial fibrillation following a rectal endoscopic injection using epiphedrine solution]. PMID- 11938054 TI - [Increased hepatocyte growth factor serum levels in patients undergoing elective surgery]. PMID- 11938055 TI - [Hepatic actinomycosis associated with gastric leiomyosarcoma. A case report]. PMID- 11938056 TI - [Chronic hepatitis with an acute presentation due to Ecstasy]. PMID- 11938057 TI - [Endoscopic and histological findings of colonic pseudo-lesions induced by Fleet Phospho-Soda (R)]. PMID- 11938058 TI - [Combined interferon-lamivudine as a first line therapy for chronic hepatitis B]. PMID- 11938059 TI - [Toward the expected development of therapeutic combinations in hepatitis B?]. PMID- 11938061 TI - [New management strategies for hepatitis B]. PMID- 11938062 TI - [Regional differences in the access to liver transplantation in France]. AB - AIM: This study evaluated regional differences in registration on the waiting list and access to liver transplantation in France. METHODS: The annual incidence of registration on the waiting list was calculated with the number of transplantation candidates registered in each region between 1996 and 1998 and was used as an indicator for access to the waiting list. The median waiting time before receiving a graft was estimated for patients registered between 1992 and 1998 by the Kaplan Meier method using transplantation as the event. This was used as an indicator for access to transplantation. Shortage of liver grafts was based on the ratio between the number of registered patients and the number of transplantations. RESULTS: The national incidence rate on the liver transplantation waiting list was 11.5 candidates per million inhabitants (pmi). There were no significant statistical differences among regions but it was high for the Rhone-Alpes region (18.6 pmi) and low for the French West Indies (3.6 pmi). The median waiting time before a graft is 2.2 months, and varied from 0.5 month to 3.8 months in French regions (p<0.001). Lack of a graft was strongly correlated with the median waiting time but not with the harvested liver rate. CONCLUSION: Although there is no statistical significant difference in access to the waiting list for liver transplantations, access to grafts differs among the regions in France. PMID- 11938063 TI - [HBs Ag and antibodies to hepatitis C virus in complicated chronic liver disease in Gabon. A case control study]. AB - BACKGROUND AND OBJECTIVE: The prevalence of HBs Ag and anti-HCV antibodies are high in the general population in Gabon. The aim of this study was to perform a case control study to determine the role of hepatitis B and C viruses in decompensated cirrhosis and hepatocellular carcinoma. METHODS: Between October 1990 and June 1998, HBs Ag and anti-HCV antibodies were investigated in 1 204 newly hospitalized patients. Sixty-seven had decompensated cirrhosis, 38 had hepatocellular carcinoma and six an association of both diseases. Prevalences were compared with those in a group of 527 sex and age matched controls from the same cohort. RESULTS: HBs Ag prevalence among cases was 35.1% (decompensated cirrhosis: 34.2%; hepatocellular carcinoma: 40.5%) and 12.5% among controls. Anti HCV were detected in 32.4% of cases (decompensated cirrhosis: 34.2%; hepatocellular carcinoma: 28.6%) and in 20.1% of controls. Complicated chronic liver disease was linked to HBs Ag (OR=11.3; IC: 4.8-26.7; cirrhosis: OR=18; IC: 5.3-61.5; hepatocellular carcinoma: OR=8.3; IC: 2.5-27.8) in patients from 15 to 34 years old. Above 45 years, complicated chronic liver disease was linked to anti-HCV antibodies (OR=2.9; IC: 1.6-5.3; cirrhosis: OR=2.8; IC: 1.4-5.8; hepatocellular carcinoma: OR=3.2; IC: 1.1-9.5). CONCLUSION: Both Hepatitis B and C viruses are linked to complicated chronic liver disease in Gabon in an age dependent manner. PMID- 11938064 TI - [The liver and myeloproliferative syndromes]. PMID- 11938065 TI - [Therapeutics of squamous cell cancer of the anus: specific aspects in HIV infected subjects]. PMID- 11938066 TI - [Anal canal squamous-cell carcinomas in HIV positive patients: clinical features, treatments and prognosis]. AB - The prevalence of squamous-cell carcinoma of the anus seems to be increasing in HIV positive patients. Clinical features and prognosis in this population have not been well evaluated. AIMS: To assess the prognosis of anal squamous-cell carcinoma in HIV positive patients as well as clinical features and treatment procedures. METHODS: A series of 20 HIV positive patients presenting with invasive anal squamous-cell carcinoma was retrospectively analyzed. Data have been compared to those obtained from 24 randomly selected HIV negative patients who were followed during the same periods in the same centers for anal carcinoma with similar histopathological features. RESULTS: The follow-up ranged from 10 to 172 months. No difference was observed between the two groups concerning the clinical features leading to anal cancer diagnosis, although HIV positive patients were younger. Anal cancer was more frequently associated with lymph node metastasis in HIV positive (60%) than in HIV negative (17%) patients, although its size was similar in both groups. Radiotherapy was similarly performed in both groups, while chemotherapy was administered less frequently in HIV positive than in HIV negative patients (54% vs 25%). Immediate side effects and mortality at 1 year follow-up were similar in both groups, whereas the objective initial response to therapy (50% versus 88%), the remission rate with anal conservation at 1 year follow-up (45% versus 88%), and the mortality at 3 years were better in HIV negative patients. CONCLUSION: The prognosis of anal squamous-cell carcinoma is poor in HIV positive patients. This correlates with a more advanced tumor stage and an alteration of systemic immunity status at the time of diagnosis and less response rate to treatment. Detection of precancerous lesions and treatment procedures should be evaluated in HIV infected patients. PMID- 11938067 TI - Epidemiology of inflammatory bowel diseases in the elderly in the province of Liege. AB - OBJECTIVE: Inflammatory bowel diseases (IBD) are heterogeneous diseases which affect preferentially young adults. The late onset could represent a particular form of expression of these diseases. The aim of our prospective study was to describe the incidence of IBD in patients older than 60 years as well as their clinical pattern in comparison with a population younger than 60. METHODS: A standardized questionnaire for each new case diagnosed in the province of Liege between 01/06/1993 and 31/05/1996 was completed. RESULTS: During the three years, 270 patients were enrolled. In group IBD > 60 years old, there were 60 new cases, including 23 cases with Crohn's disease (CD) (38%), 30 with ulcerative colitis (UC) (50%), and 7 with undetermined colitis (IC) (12%). The proportion of CD was significantly lower in the group IBD > 60 years old than in the group<60 (114 CD (54%), 81 UC (39%) and 15 IC (7%); P=0.04). The annual incidence tended to be higher for UC than for CD in group IBD > 60 (4.5 and 3.5 per 100,000, respectively) while it was the contrary in younger patients (3.4 and 4.8 per 100,000, respectively). There was no striking difference in the clinical features for both diseases in the two groups, except more frequent diarrhea, weight loss and extraintestinal symptoms in CD patients<60 years old. CONCLUSIONS: In the province of Liege, the incidence of IBD in people older than 60 years is high. IBD in the elderly is characterized by a lower proportion of CD than in the younger population. Clinical features tend to be the same whatever the age at diagnosis for each disease. PMID- 11938068 TI - [Gastric inflammatory response in Helicobacter pylori infection]. PMID- 11938069 TI - [Solitary fibrous liver tumor: clinical, radiological and pathological characteristics]. AB - A solitary fibrous liver tumor is a rare disease which is difficult to diagnose. Radiological findings are not specific, and cannot confirm benignity or malignancy. Treatment most often involves a major hepatic resection. A precise diagnosis is made by pathological examination, mainly immunohistochemistry with positive results for anti CD34 and anti vimentin antibodies. Prognosis is directly correlated to the presence of malignant histoprognostic features, i.e. the number of mitoses. We report a case of a benign solitary fibrous liver tumor, which presented as a voluminous hepatic mass with few symptoms and was treated successfully by radical hepatic resection. PMID- 11938070 TI - [Colonic malacoplakia and ulcerative colitis: report of a case]. AB - Malacoplakia is a form of chronic granulomatous inflammatory reaction that rarely affects the digestive tract and has exceptionally been reported in association with ulcerative colitis. We report a new case in a 58-year-old woman suffering from ulcerative colitis. As colitis worsened, the patient received systemic steroid therapy but symptoms did not improve. As colonic perforation was suspected, a sub-total colectomy was performed. Histopathological study revealed a diffuse infiltration of the colonic mucosa by sheets of large macrophages with eosinophilic granular cytoplasm and characteristic cytoplasmic inclusions (Michaelis-Gutmann bodies) together with active and chronic lesions of ulcerative colitis. Malacoplakia gradually disappeared under antibiotics and did not recur whereas ulcerative colitis remained active. In our case, as in three similar published cases associated with ulcerative colitis or Crohn's disease, malacoplakia was probably triggered by steroid therapy and was not clinically suspected. This particular and potentially severe inflammation must be recognized and treated in order to prevent worsening of the associated bowel disease. PMID- 11938071 TI - [Severe acute hepatitis induced by alendronate]. PMID- 11938072 TI - [Severe hemorrhage from the epigastric after ascites paracentesis]. PMID- 11938073 TI - [Phytosoya(R)-induced cytolytic hepatitis]. PMID- 11938074 TI - [Barrett's esophagus can progress to adenocarcinoma following total duodenal diversion]. PMID- 11938075 TI - [Duodenal hematoma associated with thrombopenia in chronic alcoholic pancreatitis]. PMID- 11938076 TI - [Whipple's disease with exsudative ascites]. PMID- 11938078 TI - [The Journal changes to the Euro] PMID- 11938077 TI - [Corticosteroids in the treatment of immune thrombocytopenic purpurain a patient with chronic hepatitis C virus infection]. PMID- 11938079 TI - ? PMID- 11938081 TI - [EDITORIAL] PMID- 11938080 TI - [103rd French Congress of Surgery (Paris, October 4-6, 2001)] PMID- 11938083 TI - [MULTIPLE CHOICE QUESTIONS] PMID- 11938082 TI - [Foreword] PMID- 11938084 TI - [Pathological angles] PMID- 11938086 TI - [MULTIPLE CHOICE QUESTION] PMID- 11938085 TI - [Foreword] PMID- 11938087 TI - [Metastases of the iris] PMID- 11938111 TI - [To our readers]. PMID- 11938112 TI - [Ionizing radiation and health: data and objectives]. AB - Three dimensions must be considered to analyze the consequences of exposure to ionozing radiation on public health: assessment of exposure, measure of the public health risk, and monitoring the perception of risk by the population. We reviewed these three dimension for several situations of exposure to ionizing radiation: exposure related to the Chernobyl accident, occupational exposure, military-related exposure and medical exposure. For each situation, we discuss the pertinence of available data an monitoring systems used in France, and propose when necessary new systems. An extension of the cancer registries to the entire country would be a significant progress in terms of research and surveillance. This must be discussed in terms of potential utility in other domains of public health. PMID- 11938113 TI - [In the field of ionizing radiations, are the dosimetric data satisfactory?]. AB - The quantification of the dose is a key element for the surveillance of radiological exposures of the population. The objectives are the reduction as low as reasonably achievable of exposures, the assessment of the risk in complement to epidemiologic surveys and refining the dose/effect relationship. However, two difficulties should be highlighted: first, the dose delivered into organs or to the whole organism is not directly measurable and second, its translation into risk (cancer hazard, for instance) requires precautions. The questions associated with the development of a good dosimetry depend on metrology, modeling, investigations, management of databases, and give rise to debates with regard to the notion of realism and confidentiality of individual data. This activity experiences deep changes due, not only to the development of techniques and the increasing social demand, but also to changes to the regulations, in particular with the transposition of the European Directive for the protection of the patients (97/43, June 30, 1997) and of the Directive relating to the basic safety standards (96/29, May 13, 1996). Various types of situations are distinguished: professional exposure, public exposure (releases of facilities handling radioactive materials, natural sources, contaminated sites, nuclear test fallouts) and patients'exposure (radiology and radiotherapy). Recent investigations at European or domestic level are mentioned, the purpose of which is to evidence the benefits and lacks, if any, in the dosimetric monitoring for these various types of situations. As a conclusion, orientations are defined and some of them are already applied, following the above-mentioned investigations or works. As the two main sources of exposure for the population are medical (41%) and natural irradiations (58%), the effort of the reduction should first cover these two components. With regard to natural radioactivity, the main stake is to identify the sectors with a high radon potential and to implement the associated reduction means. With regard to patients'exposure, the emphasis should concern the development in radiodiagnostic, of the means required for systematically recording a dose indicator for every radiodiagnostic examination and any radiological operation. Finally, the exposure of workers, which only represents a component of the overall population exposure, should be subject to an on-going attention with regard to data centralization and accessibility. PMID- 11938114 TI - [Epidemiological data and radiation risk estimates]. AB - The results of several major epidemiology studies on populations with particular exposure to ionizing radiation should become available during the first years of the 21(st) century. These studies are expected to provide answers to a number of questions concerning public health and radiation protection. Most of the populations concerned were accidentally exposed to radiation in ex-USSR or elsewhere or in a nuclear industrial context. The results will complete and test information on risk coming from studies among survivors of the Hiroshima and Nagasaki atomic bombs, particularly studies on the effects of low dose exposure and prolonged low-dose exposure, of different types of radiation, and environmental and host-related factors which could modify the risk of radiation induced effects. These studies are thus important to assess the currently accepted scientific evidence on radiation protection for workers and the general population. In addition, supplementary information on radiation protection could be provided by formal comparisons and analyses combining data from populations with different types of exposure. Finally, in order to provide pertinent information for public health and radiation protection, future epidemiology studies should be targeted and designed to answer specific questions, concerning, for example, the risk for specific populations (children, patients, people with genetic predisposition). An integrated approach, combining epidemiology and studies on the mechanisms of radiation induction should provide particularly pertinent information. PMID- 11938115 TI - [Are new data collection systems needed to achieve public health and research goals for the prevention of suicide?]. AB - Evaluation of suicide prevention programs, both on the local and national level, is a methodological challenge. In September 2000, France initiated a nation program for suicide prevention. But, as was noted in the Premutan report and more recently the FNORS report, very few of the hundreds of preventive measures taken annually are evaluated. In addition to this assessment challenge, new data, particularly concerning the different phases of the suicide process, are needed to design new programs, define their objectives with precision, and assure their implementation. For example, if the objective is to improve prevention in men aged 25-54 years who suffer from a state of depression, alcohol abuse, impulsive personality disorders, and life events implicating loss, the necessary information is currently unavailable or sparse. The lack of data and research in this domain can be grouped into several categories: lack of proper definition and quantification of the objectives; insufficient study of the mechanisms by which prevention measures are effective: lack of monitoring tools providing information on the rate of implementation of planned actions. These arguments demonstrate the need for collecting new information to develop a dynamic prevention program. Such data collection should be synchronized, conducted by a wide range of actors, and concern multiple actions. Without a coherent information system, it is doubtful the current impetus for the prevention of suicide can be sustained. PMID- 11938116 TI - [Quality of suicide mortality data]. AB - Prevention of suicide is a public health priority in France. Indicators of suicide mortality have been widely used to describe epidemiological situations or to evaluate public health actions. It is therefore essential to examine the quality of suicide mortality data. The purpose of this work was to identify potential biases affecting the quality of such data and their comparability between different countries as well as to determine how they can affect conclusions. Potential biases were identified by studying the characteristics of the death certificate system and analyzing the international literature on data quality. The impact of biases was assessed by analyzing the causes of "concurrent" death with suicide in the official statistics (trauma and poisoning caused in an undetermined way concerning intention and unknown causes). The proportion of suicides listed as "concurrent" causes of death, estimated from specific surveys was extrapolated to official data. This method was also used to correct the international data. Practices concerning death certificates for violent deaths vary considerably from one country to another: type of certifying physician, frequency of medicolegal investigations, frequency of autopsies, suicide definition criteria, confidentiality regulations, religious and culture context. These practical differences lead to variability in undetermined and unknown causes. The corrections made on the mortality data after taking into account for these potential biases showed that the rate of suicide determined from official data is considerably underestimated, but that sociodemographic and geographic factors of suicide change little after correction. Likewise, the order by country was similar after taking into consideration concurrent causes. A reliable evaluation of the rate of suicide for a given country is of course important. However, it is possible to characterize populations at risk and analyze the determinants of suicidal behavior without necessarily recording all suicides, as long as the declaration bias is stable. Statistical analysis of death by suicide in France shows that, despite under-reporting, the principal sociodemographic and geographic features and trends over time can be considered as valid. A series of recommendations is proposed however to improve data quality and homogeneity for death certificate reporting. Designing operational criteria for deciding when to declare suicide as the cause of death would be helpful to guide physicians who report deaths. Classical autopsies could be completed by "psychological autopsies" with friends and family of the deceased. The death certificate form could be improved to include items for complementary information favoring or not suicide. PMID- 11938117 TI - [Suicide prevention strategy and recent data on treatment of depression: obstacles encountered]. AB - Until recently, national strategies have not demonstrated their effectiveness in preventing suicide. A recent series of studies have shown that more aggressive treatment of depression in the population is associated with a significant reduction in the rate of suicide. Proof that a population-based strategy for preventing suicide can be effective should be demonstrating that an etiological factor can be identified on the population level and on the individual level and that this causal factor can be corrected on the individual level. Pertinent data can be put forward and in general demonstrate that treatment of depression is an effective means of preventing suicide on the population level. PMID- 11938118 TI - [Food-borne infections: do we need further data collection for public health action and research]. AB - The data needed for controlling and preventing food-borne infectious diseases relied so far on surveillance of the main food-borne infections (Salmonella and listeria infections) to follow trends over time and detect outbreak. However, this systematic continuous data collection was not always sensitive enough to timely detect community outbreaks and did not take into account sporadic cases of food-borne infections which represent the majority of patients (such as Campylobacter infection for example). Therefore, the true burden of food-borne infections, including morbidity, mortality and the social and economic cost could not be assessed. As the issue of food-borne infections got more social attention, the need of further epidemiological data has increased including the implementation of large population studies (active surveillance and follow up studies) associated with analytical studies in the United States (Foodnet) and in some European countries (United Kingdom, The Netherlands, Denmark.). Meanwhile surveillance and detection of outbreaks have substantially improved through the use of new typing scheme, particularly those based on molecular techniques. Their routine use, now, allows an early identification of the clonal spread of food borne bacteria. The application of automatic detection algorithm to surveillance data base has also improved the performance of outbreak detection and changes of trends. Furthermore, food-borne infection surveillance has become European which allows, by pooling of national database, to identify emergence that did not get noticed in any country. To foster the prevention of sporadic cases, the use of analytical epidemiology has become more and more frequent and has improved the knowledge of food vehicles and risk factors. The risk assessment approach has also been applied to the microbial contamination of food to answer to the increasing needs of expertise, particularly at the international level. This approach, however, needs further methodological development and the collection of extra information on the food chain. Last, but not the least, a wider public health research approach must also be considered, particularly on the social perception of food-borne risks and the cost-effectiveness analysis of the prevention measures that are taken. PMID- 11938120 TI - Foodborne infectious risks: do we need a wide system of data collection and survey? The lessons learned from the study of infectious intestinal disease in England. AB - In 2000, the United Kingdom Government's Food Standards Agency published "A report of the study of infectious intestinal disease in England". This report was the result of over a decade's endeavour and cost well in excess of 2 million pound sterling (approximately 3.3 million euros). The study originated in 1989. In response to national epidemics of foodborne infection with Salmonella enteritidis phage type 4 and Listeria monocytogenes, the Government set up the Committee on the Microbiological Safety of Food (the Richmond Committee). This committee wished to know the actual level of clinical disease in the population giving rise to the laboratory reports of gastro-intestinal pathogens in national surveillance data and recommended studies to achieve this objective. In addition, successors to the to Richmond Committee decided that it would be of value to collect information from both cases and controls, including documenting exposure to biologically plausible risk factors so that differences between the ill and the well could be identified. Enquiries were also made about the clinical course of disease, so that long term sequelae and socio-economic costs could be estimated. A full description of this enormous study is beyond the scope of this paper, which defends the personal view that resources might have been better spent on a less ambitious enquiry. PMID- 11938119 TI - [Are data collected in France sufficient for a quantitative analysis of food related infections?]. AB - International marketing of animals, animal products and food stuffs and the emergence or re-emergence of food poisoning has raised the risk of international transmission of certain infectious agents. Risk analysis, with its three components, assessment, management, and information, constitutes a new approach to state health policies. To guarantee health safety of the human, animal and plant populations, states can prohibit importation or marketing of food products if their decisions are based on open scientific evidence. Risk assessment is a structured open process evaluating available data designed to modelize transmission of the infectious agent integrating the different elements of the food chain from production to consumption. The assessment process involves four phases, The results of risk assessment depend on the pertinence and the quality of the data used for each of these four phases. Data required to implement risk assessment are unfortunately widely dispersed. The food and agriculture industry, official regulatory agencies, consumer watch organizations, and the scientific community all have useful information for risk assessment. The lack of standard methods to collect, process and diffuse data further complicates the problem. PMID- 11938121 TI - [What epidemiologic surveillance after a radiation accident? Lessons from Chernobyl]. PMID- 11938319 TI - [Cellular aspect of neuroinflammation in Guillain-Barre syndrome: a key to a new therapeutic option?]. AB - Guillain-Barre syndrome (GBS) is an acute inflammatory demyelinating neuropathy associated with long-lasting morbidity and substantial risk of mortality. The two reference treatments (plasma exchange-PE and intravenous immunoglobulins-IVIGs) do not change the functional prognosis in patients with very severe disease. Pathogenesis of GBS associates recently characterized humoral and cellular immune dysfunctions. Antibodies against nerve antigens may participate in complement activation, antibody-dependent macrophage cytotoxicity, and reversible conduction failure. Cellular immune reaction is associated with an increase of proinflammatory cytokines (for exemple TNF-alpha), a decrease of anti inflammatory cytokines (such as TGF-B1), an increase of matrix metalloproteinases (MMP-9-gelatinase B), all abnomalities favoring adhesion to and transmigration across endothelium of immune cells, a key phenomenon in GBS. Recovery of GBS is characterized by the normalization of these abnormalities. Experimental allergic neuritis (EAN), the experimental model of GBS has strikingly similar immunological abnormalities. The treatments of GBS, PE and IVIGs, mainly target the humoral component of the immune response. IFN-B is a cellular immunomodulator that inhibits antigen presentation, TNF-alpha production and binding, modulates macrophages properties. IFN-B increases anti-inflammatory T cell functions and cytokines, such as TGF-B1. IFN-B has important properties on leukodiapedesis by modulating expression of cell adhesion molecules and the MMP-9 proteinase. IFN-B has been used with success in EAN, in some patients with acute exacerbation of chronic inflammatory demyelinating polyneuropathy, and in one patient with GBS. Pathophysiology of GBS, IFN-B properties, and experimental studies support the initiation of a trial of IFN-B in GBS. PMID- 11938320 TI - [Mild cognitive impairments in the elderly: a critical review]. AB - The aging of the population leads to increased awareness of problems associated with age-related degenerative dementias. Given that these dementias are progressive in onset, many clinicians and researchers have proposed criteria that allow for the identification of older subjects manifesting cognitive impairment, but not responding to the criteria for dementia. Our knowledge of subjects with mild cognitive impairment is limited; it is, however, established that they present a high risk of developing dementia in the future. Although it is essential to increase our comprehension of their cognitive and functional decline, the study of subjects presenting mild cognitive impairment is compromised due to the existence of numerous non-converging classifications. The goal of the present article is to conduct a critical review of the different classifications of mild cognitive impairment in the elderly in order to attempt to identify the optimal criteria, allowing for a distinction to be made between subjects with mild cognitive impairment, who remain in a stable state and those whose condition evolves to a dementia. These criteria may enable us to describe a homogenuous group of individuals presenting with different rates of dementia risk factors. We present the classifications most frequently used in clinical and research settings. After listing them according to categorial, clinical or dimensional approaches, we performed a critical analysis for each one. Depending on the diagnostic criteria applied, major variations are revealed for the prevalence of cognitive impairment and the incidence of dementia. They are explained by methodological and theoretical shortcomings that we point out and discuss (e.g., reference group, lack of diagnostic criteria or exclusion criteria, high level of subjectivity). Beyond these criticisms, we discuss the challenges to be met in order to reach the optimal identification criteria. Notably, the impact of mild cognitive impairment on daily living activities should be tested with the use of more specific questionnaires/tasks. The goal of the objective definition of cognitive impairment should be to minimize subjectivity in the diagnosis. It is also suggested that sensitive cognitive measures would be used on all aspects of cognition, while recognizing and taking into account all confounding factors (e.g., age, education level). Given the nature and consequences of mild cognitive impairment, an inter-disciplinary approach is suggested (e.g., neurobiological, psychiatric, and genetic cues). A consensus on optimal diagnostic criteria is essential to propose cognitive and pharmacological treatments for the effective prevention of ementia. PMID- 11938321 TI - [Validation of the French language version of the Parkinson's Disease Questionnaire - PDQ-39]. AB - After Alzheimer's disease, Parkinson's disease (PD) is the second most frequent degenerative disease of the central nervous system. The consequences of PD at the functional, social and emotional levels warrant a better understanding the patient's perceptions as measured using a specific instrument rather than restricting the medical approach to the clinical evaluation of the motor component. In 1996, we began implementation of a project to transculturally validate the single specific instrument that had been published and was available at that time: PDQ-39. The scale consists in a 39-item questionnaire enabling determination of an overall quality-of-life score and scores for 8 specific dimensions: mobility, activities of daily living, emotional well-being, stigma, social support, cognitions, communication and bodily discomfort. Eighty-nine patients taking part in an open-label study of the safety of a combination of pergolide and dopa therapy were included and followed up on D15 and after 8 weeks. The process of "Forward-Backward" translation, conducted in close liaison with the authors, enabled semantic and linguistic validation of the French language version. The content was validated by PD experts. At baseline, the patients presented quality-of-life scores that were particularly impaired for the dimensions exploring Mobility, Emotional well-being and Bodily discomfort. The main metric properties of the scale were confirmed. The PDQ-39 scores were closely correlated with the related concepts investigated by generic scale, SF 36. The PDQ-39 scores were correlated with the "Mental and Mood Status", "Everyday Activities" and "Motor Status" dimensions determined by the UPDRS. The reliability, expressed by Cronbach coefficients alpha, showed strong consistency of the instrument, very similar to the data for the original version. In contrast to what was observed with SF-36, the scale was particularly sensitive to clinical changes. The initial results make PDQ-39 a precious tool for the optimization of management of patients presenting with PD. PMID- 11938322 TI - [Endoluminal dilatations and stenosis of symptomatic vertebral arteries]. AB - Surgical treatment of symptomatic atherosclerotic stenosis of vertebral arteries has been proposed for many years but this technique remains quite confidential due to technical difficulties and relatively high risks. Transluminal angioplasty has been proposed and we developed a simplified technique using coronary stent placement. The aim of this study was to evaluate the feasibility and efficacy of transluminal angiography with primary stenting for proximal stenosis for vertebral arteries. Eleven patients with symptomatic atherosclerotic stenosis of vertebral arteries were treated by trans luminal angioplasty with primary stent placement. Two patients, one with stenosis of the proximal vertebral artery and one with distal stenosis of the vertebral artery where only treated by transluminal angioplasty. In all cases transluminal angioplasty and stenting were feasible with restitution ad integrum of the diameter of the artery in 98 cases and with residual moderate stenose (<20%) in 5 cases. All patients were followed for more than one year, only one patient had recurrence of symptoms, but he stopped spontaneously the anti platteless drugs. Vertebro basilar symptoms disappeared completely in 12/13 cases and were improved in 1/13 cases. No restenose of the artery was observed on control (echodoppler) excepted in one case, where a tight stenose of pre vertebral sub clavian artery developed. Transluminal angioplasty for symptomatic stenosis of vertebral artery appears as a very successful technique with a low complication rate. It should be proposed in many cases of vertebrobasilar insufficiency related with tight vertebral artery stenosis. PMID- 11938324 TI - [Treatment of progressive multiple sclerosis with cyclophosphamide]. AB - Intravenous (IV) cyclophosphamide is currently used in secondary progressive (SP) and Primary progressive (PP) Multiple Sclerosis (MS) but its efficacy remains uncertain. Furthermore, it is necessary to determine which MS should be successfully treated with IV cyclophosphamide. We retrospectively investigated 111 consecutive patients with progressive MS (21 PPMS and 90 SPMS) treated in an open label fashion with IV cyclophosphamide. We analysed clinical data (gender, age, duration of progression, primary versus secondary MS). The treatment response was assessed by EDSS change after 6 months and 1 year of treatment. The annual relapse average decreased from 1.92 before treatment to 0.39 during the treatment. Age and gender did not influence response to therapy. We did not find any difference of response between PPMS and SPMS. Duration of the progressive phase in SPMS was not a predictive factor of efficacy. A better response was noted in SPMS patients with surimposed relapses than in patients without relapses during the year before treatment (p<0.05). Furthermore, the better response in SPMS patients with relapses before treatment suggests that it is necessary to treat when MS is still in an inflammatory stage. PMID- 11938325 TI - [Paraplegia disclosing an intracranial arterovenous fistula with perimedullary drainage]. AB - This case report deals with a patient who presented with a rapidly progressive myelopathy due to an intracranial dural arteriovenous fistula with perimedullar drainage (type V). We shall discuss the main clinical and radiological features of this type of fistula and its treatment from a review of the published cases. PMID- 11938323 TI - [Isolated pontine infarction: vascular anatomy helps understand clinical signs]. AB - Clinical findings of pontine infarcts are more frequently described as classical crossed-syndrome. The aim of this study was to analyse clinical presentation of isolated pontine infarcts. From the Besancon Stroke Registry, we retrospectively studied 34 patients with isolated pontine infarcts documented on MRI with a strict centro-bicommissural plane as the reference. MRI were analyzed using an arterial territories mapping of brainstem. Among our 34 pts, none developed a classical crossed-syndrome. A thorough knowledge of vascular anatomy of brainstem is necessary to understand clinical findings. Moreover, this knowledge allow to explain the rare occurrence of pontine crossed-syndrome. PMID- 11938326 TI - [Late discovery of Lafora disease: a family study]. AB - Lafora disease is a progressive myoclonic epilepsy, Clinically defined by the association of myoclonic, epileptic fits and dementia. We report a case with an atypical Lafora disease, marked by delayed onset at 25 years of age, prolonged course, associated with secondary cognitive impairment and myoclonic features. PMID- 11938327 TI - [Meningo encephalitis revealing Q fever: two cases and a review of the literature]. AB - Q fever is a zoonosis caused by Coxiella burnetii. The acute Q fever is usually characterized by a self-limited flu-like syndrome, fever, pneumonia and hepatitis. Symptoms of the chronic Q fever (evolution>3 months) mainly consist of endocarditis with negative culture. Focal neurological symptoms are rarely observed. Neurological symptoms of acute Q fever consist of meningitis or meningo encephalitis. Neurological symptoms of chronic Q fever are cerebral embolisms from Coxiella burnetii infected heart valves. We herein report two patients with meningoencephalitis revealing acute Q fever. PMID- 11938328 TI - [Acute meningomyelitis and polyradiculoneuritis disclosing systemic lupus erythematosus]. AB - A 25-year-old male presented purulent meningitis associated with transverse myelitis. Spinal T2-weighted MRI showed a large spinal cord with an intramedullary high signal. Infection resolved with antibiotic therapy but spastic paraplegia persisted. Four months later, he developed a Guillain-Barre syndrome with clinical and biological signs of systemic lupus erythematosus. Final outcome was fatal despite corticosteroid and immunoglobulin treatment. PMID- 11938329 TI - [Neuritic leprosy disclosed by reversal reaction]. AB - We report a case of neuritic borderline tuberculoid leprosy with lingering insidious growth disclosed by a brutal reversal reaction. Inflammatory polyarthralgia and sensory and motor loss in the median and ulnar territories, without skin lesions, suggested vasculitis. A few weeks later, inflammatory skin lesions developed leading to the diagnosis of biopsy proven leprosy. Leprosy should be considered as a possible diagnosis in patients with multineuritis, especially when associated with rheumatic or cutaneous manifestations. Early skin biopsy in neuritic leprosy is discussed. PMID- 11938332 TI - [Digestive tract disorders revealing mitochondrial cytopathy: MNGIE syndrome]. PMID- 11938333 TI - [Acute disseminated encephalomyelitis]. PMID- 11938341 TI - Endovascular treatment of aortic aneurysms. PMID- 11938343 TI - The role of the vascular nurse in centers of excellence. PMID- 11938344 TI - Dollars and sense: the economics and outcomes of patients undergoing carotid endarterectomy at Royal Adelaide Hospital. AB - This study examined the costs and outcomes of patients undergoing carotid endarterectomy who returned directly to the vascular unit after surgery rather than to the intensive care unit (ICU)/high dependency unit (HDU). The ICU/HDU is for critically ill patients. HDU is the step-down area from the ICU. The nurse-to patient ratio for these patients is 1:2 and senior medical staff are available 24 hours a day. To prepare staff for the change in practice, an intense educational program was provided and protocols for patient management were developed and implemented. Outcomes were then monitored for the group of patients who returned directly to the vascular unit. Case notes audits, informal patient interviews, daily monitoring of patient outcomes, and an analysis of costing data from Australian Diagnostic Related Groups demonstrated that 50% of patients required transfer to the ICU/HDU after surgery for respiratory, hemodynamic, or neurologic management. The remaining 50% of patients returned directly to the vascular unit with no major complications and had better outcomes with less intensive nursing care overall. The length of hospital stay and the management costs were reduced significantly. PMID- 11938345 TI - Development and implementation of a critical pathway for abdominal aortic aneurysms in Japan. AB - Although the critical pathway (CP) has been implemented successfully for simple procedures and recovery processes, yielding improvement in length of stay (LOS) and patient satisfaction, there are few reports about its application to complicated procedures and recovery processes in Japan. A model was developed of the complicated postoperative abdominal aortic aneurysm (AAA) recovery process from 3-year patient data, including statistics such as days for achieving outcomes, comorbidities, complications, and patient characteristics. Its accuracy has been investigated by comparing patient outcomes. Eighty-three patients were divided into 2 groups (typical and atypical) on the basis of select criteria and analyzed by correlation coefficient, multiple regression analysis, and logistic analysis. No risk factors for delay of the recovery and the adverse outcomes were detected. The CP was the wound healing process in the short-term recovery process. The average LOS was 36.3 days in the typical group and 47.2 days in the atypical group. Stitch removal occurred 8 days after surgery, and patients were discharged a minimum of 11 days after the operation. The AAA-CP has been implemented since September 1998 in 45 experimental cases, and a significant reduction in days required to achieve short-term outcomes has been observed (25.6 days to 19.1 days in LOS after surgery) as well as a decrease in the prevalence of adverse outcomes. PMID- 11938346 TI - Controlled, randomized clinical trial of 2 hydrocolloid dressings in the management of venous insufficiency ulcers. AB - A prospective, randomized study was conducted to compare the performance of 2 hydrocolloid dressings, hydrocolloid A and hydrocolloid B, in the treatment of venous insufficiency ulcers. A total of 31 patients were enrolled at 2 clinical sites. Complete wound closure (100% epithelialization) was observed in 59% of the patients treated with hydrocolloid A, compared with complete wound closure in 15% of the patients in the hydrocolloid B group (P 5' exonuclease that catalyzes the deletion of nucleotides at coding joins. These findings suggest that the two TdT isoforms may act in concert to preserve the integrity of the variable region of antigen receptors while generating diversity. PMID- 11938352 TI - The mirrored methionine sulfoxide reductases of Neisseria gonorrhoeae pilB. AB - Methionine sulfoxide reductases (Msr) protect against oxidative damage that can contribute to cell death. The tandem Msr domains (MsrA and MsrB) of the pilB protein from Neisseria gonorrhoeae each reduce different epimeric forms of methionine sulfoxide. The overall fold of the MsrB domain revealed by the 1.85 A crystal structure shows no resemblance to the previously determined MsrA structures from other organisms. Despite the lack of homology, the active sites show approximate mirror symmetry. In each case, conserved amino acid motifs mediate the stereo-specific recognition and reduction of the substrate. Unlike the MsrA domain, the MsrB domain activates the cysteine or selenocysteine nucleophile through a unique Cys-Arg-Asp/Glu catalytic triad. The collapse of the reaction intermediate most likely results in the formation of a sulfenic or selenenic acid moiety. Regeneration of the active site occurs through a series of thiol-disulfide exchange steps involving another active site Cys residue and thioredoxin. These observations have broad implications for modular catalysis, antibiotic drug design and continuing longevity studies in mammals. PMID- 11938353 TI - A protein sequence that can encode native structure by disfavoring alternate conformations. AB - The linear sequence of amino acids contains all the necessary information for a protein to fold into its unique three-dimensional structure. Native protein sequences are known to accomplish this by promoting the formation of stable, kinetically accessible structures. Here we describe a Pro residue in the center of the third transmembrane helix of the cystic fibrosis transmembrane conductance regulator that promotes folding by a distinct mechanism: disfavoring the formation of a misfolded structure. The generality of this mechanism is supported by genome-wide transmembrane sequence analyses. Furthermore, the results provide an explanation for the increased frequency of Pro residues in transmembrane alpha helices. Incorporation by nature of such 'negative folding determinants', aimed at preventing the formation of off-pathway structures, represents an additional mechanism by which folding information is encoded within the evolved sequences of proteins. PMID- 11938354 TI - Origins and kinetic consequences of diversity in Sup35 yeast prion fibers. AB - A remarkable feature of prions is that infectious particles composed of the same prion protein can give rise to different phenotypes. This strain phenomenon suggests that a single prion protein can adopt multiple infectious conformations. Here we use a novel single fiber growth assay to examine the heterogeneity of amyloid fibers formed by the yeast Sup35 prion protein. Sup35 spontaneously forms multiple, distinct and faithfully propagating fiber types, which differ dramatically both in their degrees of polarity and overall growth rates. Both in terms of the number of distinct self-propagating fiber types, as well as the ability of these differences to dictate the rate of prion growth, this diversity is well suited to account for the range of prion strain phenotypes observed in vivo. PMID- 11938356 TI - Hepatitis C virus genotypes in Saudi Arabia. AB - Hepatitis C virus genotypes have been associated with specific geographical areas and in many cases with specific mode of transmission. In developed countries, genotype determination has formed a part of the management of patients with hepatitis C virus seropositivity and liver diseases due to hepatitis C virus. The epidemiology of hepatitis C virus has been shown to be changing rapidly in many countries due to population movement and different life-styles; hence the distribution of the genotypes is being monitored closely in many countries. In the Kingdom of Saudi Arabia, there are only a handful of publications recording the hepatitis C virus genotypes in various population groups. These studies have been carried out mainly in Riyadh (Central province) and Jeddah (Western province). There are no studies emanating from the Eastern or Northern provinces. According to these studies, the most prevalent genotype in the Western Province and probably in the whole Kingdom of Saudi Arabia was genotype 4, followed by genotypes 1a and 1b. Genotypes 1, 2a,/2b, 3 and 6 are very rare in the Kingdom of Saudi Arabia. Genotype 5 was identified exclusively in the Western province and nowhere else. Genotypes 1b and 4 were associated with different histological grades of liver disease. Mixed infections with more than one genotype were observed in some studies. More detailed epidemiological studies of hepatitis C virus infections are needed in the Kingdom of Saudi Arabia to gain more insight into a possible type/subtype-specific pathogenesis of hepatitis C virus in the different regions of the Kingdom of Saudi Arabia as well as the distribution of the genotypes in the various localities. PMID- 11938357 TI - Recombinant tissue plasminogen activator for acute ischemic stroke. AB - Stroke is a leading cause of serious and long-term disability and death worldwide, with approximately 750,000 strokes occurring annually in the United States of America. The risk of stroke doubles each decade for people over 55 years. Cerebral angiography conducted soon after the onset of stroke demonstrates arterial occlusion in 70%-80% of cases. Recanalization of an occluded cerebral artery may assist in the recovery of reversibly ischemic tissue and limit the neurological disability. In June 1996, the recombinant tissue plasminogen activator was approved as a safe and an effective intravenous treatment for acute ischemic stroke, especially if given within 3 hours of onset of symptoms. Since approval, less than 5% of all stroke patients are receiving recombinant tissue plasminogen activator. In this review we try to answer the question of whether recombinant tissue plasminogen activator therapy should be the first-line treatment for acute ischemic stroke. The result of major recombinant tissue plasminogen activator trials will be summarized and reviewed critically. PMID- 11938358 TI - Biomedical research in the Kingdom of Saudi Arabia (1982-2000). AB - OBJECTIVE: Because of the leading position of the Kingdom of Saudi Arabia in biomedical research among the Arab States, we aimed this study at providing a detailed analysis of the development of this domain in the Kingdom. METHODS: More than 1500 queries were mainly directed to the PubMed bibliographic database. Tags, boolean operators, and wild cards were utilized. RESULTS: Most of the Saudi biomedical citations originated from the capital city of Riyadh (70%), particularly from the King Saud University (29.5%) and King Faisal Specialist Hospital (21.5%). One of every 5 Saudi papers either describes a disease, a blood related analysis, or surgical observations. Only 0.5% of Saudi papers appear in the 200 highest impact-factor journals. CONCLUSION: A steady advancement in biomedical research is observed in the Kingdom mainly in years 1987-1996. The absence of uniformity in writing the addresses of authors in published articles from Saudi Arabia results in misleading data about the geographical distribution of biomedical research and sometimes improper rating of the scientific institutions in the Kingdom. The most dynamic biomedical fields in the Kingdom observed in the present analysis are Hematology, Surgery, Cancer, Pharmacology, and Microbiology. The main reasons behind the small number of Saudi papers appearing in high impact-factor journals are the bias and the economics of scientific publishing. We hope that the present study will lay the foundation for more detailed investigations on the directions of biomedical research in the Kingdom and will be of great importance to better found future strategies in this important realm. PMID- 11938359 TI - Long-term follow up of carbonic anhydrase II deficiency syndrome. AB - OBJECTIVE: To describe the long term clinical, biochemical and radiological features of 35 Saudi Arabian children with carbonic anhydrase II deficiency syndrome who have been followed at King Faisal Specialist Hospital and Research Center, Riyadh since 1979. METHODS: The records of these patients were retrospectively evaluated. The diagnosis was based on the clinical and the radiological evidence of the disease. Carbonic anhydrase II level was measured in 9 patients. RESULTS: Clinically, these patients had typical facial features, growth failure and varying degrees of psychomotor retardation. Biochemically, all children had renal tubular acidosis that was of distal type in the majority of them. Radiologically, this syndrome was characterized by metyphyseal osteopetrosis and intracranial calcification that was progressive in 2 patients. Five patients were blind secondary to optic nerve entrapment and 2 patients developed anemia and secondary erythropoesis due to bone marrow involvement. Nineteen patients had attained the final adult height; the mean adult height was 146 cm (-3 standard deviation) in 11 females and 152 cm (-4 standard deviation) in 8 males. Two patients were married and had clinically and radiologically normal children. CONCLUSION: The syndrome of carbonic anhydrase II deficiency is usually benign in nature and compatible with long term survival, however it can progress and involve the cranial nerves. Close clinical and neurological assessment of these patients is mandatory to early detect and manage potential serious complications. PMID- 11938360 TI - Right lower quadrant pain in females. Is it appendicitis or gynecological? AB - OBJECTIVE: To determine if a gynecological consultation is needed for patients who are labeled to have acute appendicitis. METHODS: A retrospective study carried out in Assir Central Hospital, Abha, Kingdom of Saudi Arabia. Fifteen female patients who were clinically diagnosed and operated upon for acute appendicitis were discovered intra-operatively to have unrelated gynecological lesions, are presented. RESULTS: Histopathological reports on the removed appendices revealed 80% "normal" and 20% "mildly inflamed". Seven (46.7%) of the patients had right ovarian cysts (one of them, bilateral); 4 (26.7%), corpus luteum cysts, and 4 had bilateral salpingitis, bilateral pyosalpinx, right ovarian cyst with bilateral salpingitis and ruptured right tubal pregnancy each. The majority (60%) of the women were in the 20-30-year-age bracket. CONCLUSION: The need for gynecological review of female patients of childbearing age presenting with lower abdominal pain is stressed. Ultrasonography is an important adjunct in improving diagnostic accuracy in such cases. PMID- 11938361 TI - Craniopharyngioma. Analysis of factors that affect the outcome. AB - OBJECTIVE: The aim of this work is to find out factors that affect the outcome of treatment of patients with craniopharyngioma treated at King Khalid University Hospital, Riyadh, Kingdom of Saudi Arabia. METHODS: This study was carried out retrospectively by reviewing the medical records of all patients with craniopharyngiomas treated at King Khalid University Hospital, Riyadh, Kingdom of Saudi Arabia in the last 10 years (January 1990 through to December 1999), and collecting clinical, radiological, surgical, and follow-up data and performing statistical analysis to find out factors that affect the outcome of such cases. RESULTS: There were 11 females (61%), and 7 males (39%), the median age was 24 years. The main presenting symptoms were reduced vision in 66.7%, symptoms of raised intracranial pressure in 50%, endocrinal problems in 33.3%, seizures in 16.7%, and hemiparesis in 5.6%. Radiologic studies showed 72% of tumors had mixed solid and cystic components, calcification in 83%, and ventricular dilatation in 50% of cases. The tumor extended to the posterior fossa in 11%, and to both middle and posterior fossae in 11% of cases. Eighty-nine percent of patients had tumor excision through pterional craniotomy and 11% through subfrontal approach. Gross total removal was achieved in 17% and subtotal resection in 83%. Tumor recurred in 9 patients (50%), 4 of them (44%) had postoperative radiotherapy. The outcome was good in 10 patients (56%), poor in 6 patients (33%), and 2 patients died (11%). The patient age, radiological appearance of tumors and their location were significantly correlated with the outcome (p 0.02, 0.02, 0.04). CONCLUSION: Ophthalmologists and Pediatricians should be aware of the clinical presentation and refer patients to specialized centers for treatment. Total resection of the tumor should be the goal of the Neurosurgeon as it offers the best chance of cure. PMID- 11938362 TI - Outcome of thymectomy in patients with Myasthenia Gravis. AB - OBJECTIVE: Myasthenia Gravis is a neuromuscular disorder characterized by weakness and fatigability of skeletal muscles. Thymectomy is now beneficial even for non-thymomatous patients. The aim of this paper is to evaluate the role of thymectomy as on the treatment options for myasthenia gravis. METHODS: Thirty patients underwent thymectomy in the period from April 1999 through to July 2000. They were collected from different teaching hospitals in Baghdad, Iraq. All these cases were studied conventionally including history, physical and neurological examination in addition to investigations with particular emphasis on connective tissue screen and thyroid function test. RESULTS: The age in this study ranged between 17-55 years. The preoperative duration ranged between 2-20 months and the postoperative follow up period ranged between 6 months-14 years. The most favorable results were obtained in patients without thymoma with short duration. CONCLUSION: Early thymectomy carries the best results in Myasthenia Gravis and every effort should be made to shorten the preoperative duration for both thymomatous and nonthymomatous patients. Patients with symptoms can be better controlled with anticholinesterase drugs than patients using steroid and cytotoxic drugs. Thymectomy is beneficial for most patients with Myasthenia Gravis and especially in those with benign folicular hyperplasia. PMID- 11938363 TI - Psychiatric admission in a general hospital. Patients profile and patterns of service utilization over a decade. AB - OBJECTIVE: To analyze the socio-demographic and clinical characteristics of admitted patients and the patterns of their service utilization over a decade from March 1988 to March 1998. METHODS: Prospective data compilation using a structured questionnaire, hospital records and follow-up observations at King Fahd Hospital of the University in Al-Khobar, Kingdom of Saudi Arabia. RESULTS: A total of 1366 patients (683 of each sex) had 2217 admissions in 10 years. By the International Classification of Diseases, 10th edition criteria, 19.5% had schizophrenia, 15.2% bipolar disorder, 9.9% depressive episodes, 8.6% acute and transient psychotic disorders, 7.7% adjustment disorders and 7.6% dissociative disorders. Males were more frequently admitted for schizophrenia and females for mood and anxiety disorders. Most non-Arab expatriates were diagnosed as acute and transient psychotic, stress-related or dissociative disorders. Re hospitalizations constituted 28% of all admissions. The mean length of stay was 25 days per admission and 41 days per patient. The overall bed occupancy rate was 84.9%. A subgroup of 16.9% of patients, mostly with schizophrenia or bipolar disorder, consumed 62.3% of the bed occupancy. CONCLUSION: Gender and immigration were the main determinants of variance in patient characteristics, nosological distribution and pattern of service use. More beds are needed. Psycho-educational programs should be intensified to reduce the social stigma and societal intolerance to mental patients. Active family involvement improves compliance and might reduce re-hospitalization rates. Heavy service consumers should be transferred to long-stay facilities. PMID- 11938364 TI - Impact of a mini-clinic on diabetic care at a primary health care center in southern Saudi Arabia. AB - OBJECTIVE: The aim of this study was to assess the impact of a mini-clinic on the quality of diabetic care at a Primary Health Care Center in Aseer region, Kingdom of Saudi Arabia. METHODS: All the files of diabetics in Wasat Abha Primary Health Care Center were reviewed at the end of 1997 for diabetic process based on a scoring system of 11 items. Diabetic outcomes were evaluated in accordance with Quality Assurance Protocol. Data of all the files was entered into and analyzed by Statistical Package for Social Sciences. Relevant statistical tests were used. RESULTS: Files of 198 patients were evaluated, 61.6% were male, 90.4% were married, and 50% were educated. The mean duration of diabetes was 7.1 years. All the 11 items of diabetic process improved significantly except for measuring blood pressure, weight and cholesterol. However, the mean of the total score increased significantly from 5.7 points to 8.2 points (P=0.00). The measured diabetic outcomes improved significantly for the provision of diabetic card, health education pamphlets, diabetic control and obesity. Ten percent of the diabetics were found to have at least one complication. Diabetic retinopathy (8.4%), impotence (8.2%), and cardiovascular (3.6) were the most prevalent recorded complications. CONCLUSION: Establishment of diabetic mini-clinic at Wasat Abha Primary Health Care Center improved the process and the outcomes of diabetic care. Further large and countrywide studies are suggested to evaluate the cost-effectiveness of such types of clinics on diabetic care. PMID- 11938365 TI - Detection of genital colonization of group B streptococci during late pregnancy. AB - OBJECTIVE: To detect group B streptococcal carrier state of Saudi females during 3rd trimester of pregnancy and to assess type of specimens and the techniques used for the organism detection. METHODS: A total of 867 consecutive vaginal and rectal swabs were obtained from 217 pregnant women at > 28 weeks of gestation and their follow up testing from King Khalid University Hospital, Riyadh, Kingdom of Saudi Arabia. Swab-specimens were cultured comparatively on Islam and Edwards blood agar plates, and into selective Lim broth. Enrichment Lim broth cultures (>12 hours) with and without positive modified coagglutination test were then subcultured on Islam and Edwards sheep blood agar plates. Presumptive colonies were then tested for group B streptococcus identity by convential biochemical reactions, serogrouping and serotyping. Collected neonatal swab-specimens (184) were also treated similarly. RESULTS: In comparison to Lim broth enrichment culture, the direct swab specimen culture on Edwards blood agar or Islam agar plates technique revealed 84% sensitivity and 100% specificity, whereas modified coagglutination test after selective Lim broth enrichment revealed 100% sensitivity and 96% specificity. Group B streptococcus was isolated in at least one of the specimens from the 217 patients in 66 cases. Of these 66 cases, group B streptococcus was isolated from both vaginal and rectal swabs in 33 (50%) cases and only from vaginal swabs in 22 (33%) and rectal swabs in 11 (17%) cases. Of the group B streptococcus positive cases, 10 (15%) cases had spontaneously lost their carriage, upon follow up testing, whereas out of the 151 negative cases, 4 (2.6%) cases became positive for group B streptococcus colonization upon follow up testing with an overall carriage rate of (60/217) 27.6%. Certain demographic factors were found to alter such rate of carriage. Additionally, 50% of group B streptococcal colonized mothers vertically transmitted the homologous serotypes of the organism to their newborns, but clinical infection was not recorded during the study period. CONCLUSION: Group B streptococci colonization rate among term Saudi pregnant women is relatively high (27.6%); and thereby constitutes a group of women whose infants are at great risk of early-onset invasive disease. The modified coagglutination test after growth amplification seems rapid and cost effective to detect lightly or heavily group B streptococcal colonized women. Vaginal and rectal swab specimens at late pregnancy appeared necessary to accurately identify group B streptococcus maternal colonization. PMID- 11938366 TI - Current concepts in gastric cancer surgery. AB - OBJECTIVE: Current problems in gastric cancer surgery concern the extent of gastric resection, the need for abdominal evisceration, the degree of lymphadenectomy, and an optimal preoperative tumor staging procedure. METHODS: A retrospective clinical trial of 284 patients who underwent surgery at Ernst Moritz-Arndt-University, Greifswald, Germany for gastric cancer between 1987 and 1996. Main outcome measures consist of epidemiological parameters, data on type of surgery, histopathology, postoperative complications, mortality and cancer survival. Statistical analysis between groups was performed using Chi square test (perioperative risk factors, tumor localization, and surgical treatment) and Mann Whitney U tests (Lauren classification). Survival was calculated according to the Kaplan Meier method. RESULTS: The results are in favor of subtotal gastrectomy performed for all T stages located in the distal or middle 3rd provided that a tumor-free margin of 5 cm in intestinal type and 10 cm in diffuse Lauren's type tumor can be achieved, since this operation carries the lowest postoperative risks and provides the best postoperative quality of life. Resection of adjacent organs are indicated only if they are invaded by the primary tumor (T4). They should not be resected as part of an extended lymphadenectomy procedure. The primary tumor site should guide the degree of lymph node removal. Multimodal therapeutic approaches and high postoperative morbidity and mortality after exploratory laparotomy justify the use of diagnostic laparoscopy in T3 and T4 stage tumors and if diagnostic scans suggest tumor spread. CONCLUSION: Even though surgery for gastric cancer is well standardized, a tailored surgical approach to different extents of gastric cancer appears justified. PMID- 11938367 TI - Utilization of emergency services in a community hospital. AB - OBJECTIVE: Over utilization of emergency room services by patients with non urgent complaints is a global problem. It results in a waste of resources, stress among the emergency room staff and an increase in waiting time for patients requiring attention. This study was carried out to establish the extent of inappropriate emergency room attendance in a Saudi community. METHODS: Data was collected from the emergency room register, regarding the age, sex, presenting complaints, time of presentation and disposal of patients, from January 1st 1999 to March 31st 1999 at Al-Kharj Military Hospital, Al-Kharj, Kingdom of Saudi Arabia. Patients were classified in different categories according to the triage criteria of the hospital. EPI-INFO statistical software was used for calculating x(2) and p vales. RESULTS: Among the 3928 patients, 2183 (55%) were males and 1745 (44.4%) were females, while 2335 (59.4%) of the patients had primary care or non-urgent problems. In both males and females categories, 21% of the patients had respiratory tract infection followed by miscellaneous complaints like mild conjunctivitis, allergic rash, represcription for medications, minor burns (500, 12.7%, x(2) = 97.49, p < 0.00001), gastrointestinal tract problems (434, 11%, x(2) = 146.55, p < 0.00001) and aches and pains (304, 7.7%, x(2) = 283.39, p < 0.00001). In male and female categories the 2nd most common complaints were trauma (487, 22.3%) and obstetrics and gynecological problems (325, 18.6%). The majority of the patients, 1806 or 46%, attended the emergency room during night shift (2300 hours-0700 hours). Referral rates for male and female patients were 211 (9.6%) and 331 (18.9%). CONCLUSION: Similar to the findings of other nations, inappropriate utilization of the emergency room is a big problem in the Saudi community. The majority of the patients come with minor self-limiting complaints. Maximum rush was seen at night time. There is a need for health education of such groups of patients as well as finding alternative solutions. PMID- 11938368 TI - A modified method of submental oroendotracheal intubation. AB - OBJECTIVE: The aim of this paper is to present a modification of submental intubation in the management of airway. METHODS: Following usual oroendotracheal intubation, the tube and cuff are exteriorized through a laterosubmental incision, and connected to the anesthetic machine. RESULTS: Ten patients comprising 8 panfacial fractures and 2 orthognathic surgical cases have successfully been managed with this approach. There were no complications, no morbidity, and the scar was inconspicuous and esthetically pleasing. CONCLUSION: The laterosubmental intubation technique is simple, efficacious in the management of severe maxillofacial injuries, orthognathic surgical cases with nasal obstruction and in plastic surgery for concomitant approach to rhytidectomy, lip correction and rhinoplasty. This approach deserves to be widely propagated among specialists working on the face. PMID- 11938369 TI - Caries prevalence in Saudi primary schoolchildren of Riyadh and their teachers' oral health knowledge, attitude and practices. AB - OBJECTIVE: The purpose of the present study was to determine the caries prevalence and severity in primary school children Riyadh, Kingdom of Saudi Arabia, and to assess the oral health knowledge, attitude and practices of their teachers. METHODS: A random sample of primary schoolchildren was examined for dental caries utilizing World Health Organization criteria for diagnosis of caries. The information regarding oral health knowledge, attitude and practices in schoolteachers was collected through an especially designed self-administered questionnaire. RESULTS: The prevalence of caries among the sample was 94.4%. The mean decayed, missing and filled score for primary teeth of the sample was 6.3 (+3.5), with decay component of 4.9 (+3.1), missing component of 1.1 (+1.7) and filled component of 0.3 (+1.0). The mean decayed, missing and filled score for permanent teeth of the sample was 1.6 (+1.5) with decay component of 1.5 (+1.4) as the major component. All the teachers (100%) thought that good dental health was important for general health and that routine check up dental visits help in maintaining good dental health. All the teachers (100%) thought that tooth cleaning using brush or miswak was important for good dental health. Regarding the caries risk factors, the majority of teachers understood the main causes of dental caries such as poor oral hygiene (97.4%) and high sugar intake (97.4%). Almost all the teachers (97.4%) thought that dental health lectures were of great importance for their students but less than half (41%) actually spent time on teaching proper oral health care. CONCLUSION: The caries prevalence in Riyadh primary schoolchildren is very high. The teachers' knowledge regarding oral health is satisfactory and their attitude towards oral health is very positive. PMID- 11938370 TI - Effect of dietary zinc deficiency on rat lipid concentrations. AB - OBJECTIVE: Evaluation of the lipid profile in serum, liver, and testis of rats fed marginal and severe zinc deficient diets. METHODS: Three groups of rats were treated for 8 weeks with normal diet, marginally zinc deficient diet and severely zinc deficient diet. Lipid concentrations were measured in serum, liver, and testis of these groups. RESULTS: The concentrations of serum lipids were not significantly altered between marginally zinc deficient diet treated and control rats. However, in rats treated with severely zinc deficient diet, the concentrations of serum total cholesterol, high density lipoprotein cholesterol and phospholipids were significantly increased (P < 0.01) and (P < 0.001), whereas the concentration of triacylglycerol was significantly decreased (P < 0.01). However, low-density lipoprotein cholesterol concentration was non significantly different from controls. The concentrations of liver total cholesterol, triacylglycerol and phospholipids were significantly decreased (P < 0.001) in rats treated with severely zinc deficient diet. The testicular concentration of total cholesterol was increased but this increase was non significantly different from controls, whereas the testicular concentrations of triacylglycerol and phospholipids were significantly decreased (P < 0.001) in rats treated with severely zinc deficient diet. CONCLUSION: These results suggest that a marginally zinc deficient diet does not play a significant role in altering rat lipid concentrations. However, the changes in serum lipid concentrations could be related to those changes in tissue lipid concentrations. PMID- 11938371 TI - Prevalence of systemic lupus erythematosus in central Saudi Arabia. AB - OBJECTIVE: Although systemic lupus erythematosus of varying severity has been seen in our clinics, there is no information regarding the prevalence of the disease among Saudi nationals. We conducted this study to determine the prevalence of systemic lupus erythematosus in Al-Qaseem region of the Kingdom of Saudi Arabia. METHODS: A community survey of towns and villages in the Al-Qaseem area of central Saudi Arabia was conducted in 3 phases to determine the prevalence of systemic lupus erythematosus in the region. RESULTS: Of the 10,372 studied, 2 cases of systemic lupus erythematosus were identified using the criteria set for the diagnosis of systemic lupus erythematosus by the American College of Rheumatology. Based on that, the prevalence of systemic lupus erythematosus was estimated to be 19.28 per 100,000 population in the region. CONCLUSION: The estimated prevalence of systemic lupus erythematosus in Al-Qaseem area is similar to that found in western countries. PMID- 11938372 TI - Histopathological and immuno-histochemical characteristics of primary renal primitive neuroectodermal tumor. AB - We report a 32-year-old male who presented with huge (17 x 10.5 x 5 cm) right kidney with metastasis in the liver and retroperitoneal lymph nodes. Histological, detailed immunohistochemical studies and electron microscopic examinations were performed. Microscopy revealed small to intermediate sized cells with hyperchromatic nuclei, scanty cytoplasm, abundant mitosis with no pseudorossete formation. Immunohistochemical study revealed positive staining of the tumor cells for S100, neurofilaments, neuron specific enolase, vimentin and myoglobin. Primitive neuroectodermal tumors are rare malignant round cell tumors of the kidney. A correct diagnosis can be made on light microscopic features, and by immunohistochemically positive staining for more than one neural marker. This neoplasm should be differentiated from other renal neoplasms composed of small round cells. PMID- 11938373 TI - Primary ovarian pregnancy. AB - Primary ovarian pregnancy is a rare type of ectopic pregnancy, which is difficult to diagnose clinically and even intra operatively. Here we present a case of a 20 year-old woman with ruptured right ovarian pregnancy and dermoid cyst of left ovary. PMID- 11938374 TI - Psoas abscess presenting with femoro-popliteal vein thrombosis. AB - Psoas abscess is an uncommon condition with vague clinical presentation. It generally has an insidious onset and before the advent of computed tomography, few cases were reported in the medical literature. We report the case of a middle aged diabetic woman who presented with left leg swelling. Doppler ultrasound revealed thrombosis of the popliteal vein and a collection in the left groin. Computed tomography confirmed the presence of a large left iliopsoas abscess extending to the anterior compartment of the thigh complicated with thrombosis of the superficial femoral and popliteal veins. We suggest that an iliopsoas abscess should be excluded when an immunocompromised patient presents with deep vein thrombosis. PMID- 11938375 TI - Combined brucella melitensis and streptococcal viridans endocarditis. AB - Infectious endocarditis is a potentially lethal inflammation of the hearts' inner lining invaded by microorganisms. The mortality from this illness increases as the number of infective organisms rises to 2, due to involvement of the left side of the heart. These microorganisms usually arise from the patient's own flora but can be acquired from the environment. Fever and heart murmurs are the principal clinical manifestations followed by a plethora of peripheral signs due to dissemination of microorganisms via the bloodstream. Echocardiographic imaging and sensitive culture techniques form the cornerstone of diagnosis. We report a patient with rheumatic heart disease who had combined brucella melitensis and streptococcus viridans endocarditis complicated by heart failure and an aortic root abscess. He was diagnosed on the basis of a history of prolonged fever and occupational risk as a shepherd, the presence of heart murmurs, positive blood cultures and echocardiographic evidence of aortic vegetations. He had an excellent response to intravenous antibiotic therapy combined with aortic valve replacement, which nowadays is regarded as the safest therapeutic approach for aortic valve endocarditis. PMID- 11938377 TI - Unstable angina in a young patient. PMID- 11938376 TI - Acute eosinophilic pneumonia simulating severe community acquired pneumonia. AB - Idiopathic acute eosinophilic pneumonia is a recently described cause of acute respiratory failure. It usually affects young healthy individuals. Usually non pulmonary organs are not involved. Bronchoalveolar lavage eosinophilia is required for diagnosis. This disease responds uniformly to a short course of corticosteroids and does not recur. We report a young man who presented with a 2 day history of acute respiratory failure simulating severe community acquired pneumonia and necessitating mechanical ventilation. The diagnosis was made based on the classical clinical presentation and bronchoalveolar lavage and peripheral blood eosinophilia. Acute eosinophilic pneumonia in our patient was associated with acute hepatitis and erythema multiforme. To our knowledge, this is the first time to report such an association. The case is reported with review of the literature. PMID- 11938378 TI - Patient satisfaction of the Obstetric and Gynecology Unit at King Khalid University Hospital, Riyadh, Kingdom of Saudi Arabia. PMID- 11938379 TI - Eating patterns and prevalence of obesity among Arabs living in Ahwaz, Iran. PMID- 11938380 TI - Successful pregnancy after round spermatid microinjection. PMID- 11938381 TI - Etiology of end-stage renal disease in Najran, Kingdom of Saudi Arabia. PMID- 11938382 TI - Going beyond the curriculum to promote medical education and practice. PMID- 11938383 TI - Insulin therapy for diabetic ketoacidosis. PMID- 11938384 TI - Urinary tract infection. PMID- 11938385 TI - Variations in the subclavian-axillary arterial system. AB - OBJECTIVE: The objective of this study is to report on the subclavian axillary arterial system variations observed in the dissecting room. METHODS: The morphological study of 106 formalin fixed upper limbs of adult human cadavers of both sexes was carried out at human anatomy laboratory of College of Medicine, King Saud University, Riyadh, Kingdom of Saudi Arabia over a 5 year period, to observe the distribution pattern of the subclavian axillary arterial system. RESULTS: The arterial variations in the upper extremities are quite common and may occur at the level of the thoracic outlet, axilla, arm, cubital fossa and the hand. In the present study, the anomalous topographic pattern of the subclavian axillary arterial system was revealed in 7.5% of the cadavers, whereas 92.5% exhibited classical pattern of the regional arterial anatomy. The study showed: 1. An aberrant right subclavian artery arising from the arch of aorta, distal to the left subclavian artery. 2. A bilateral common subscapular-circumflex humeral trunk (3.8%) emerging from the 3rd part of the axillary artery (branching into the circumflex humeral and thoracodorsal arteries. 3. A bilateral thoracohumeral trunk arising from the 2nd part of the axillary artery (1.9%) and branching into the lateral thoracic, circumflex humeral, subscapular and thoracodorsal arteries. These anomalies were accompanied by anomalous insertion of flexor digitorum superficialis muscle, a bilateral digastric muscle formed by the abductor pollicis longus and brevis, and an anomalous formation of median nerve by 3 roots. CONCLUSION: The aberrant right subclavian artery associated with right non recurrent laryngeal nerve and variant branching pattern of the axillary artery are of interest to anatomists, surgeons and radiologists and suggested that these anomalies must be evaluated pre-operatively. PMID- 11938386 TI - Tissue factor: biological function and clinical significance. AB - Tissue Factor is the principal cellular initiator of normal blood coagulation. It is frequently encrypted in the plasma membrane of cells in contact with blood, but under certain pathological conditions endothelial cells, monocytes or macrophages may express tissue factor; and hence trigger coagulation activation. Aberrant expression of tissue factor by these cells is thought to be responsible for the thrombophilia found in septic shock, atherosclerosis and cancer. Tissue factor is produced by tumor-associated macrophages where it is believed to play an important role in tumor growth and dissemination. It may also be involved in other cellular processes such as intracellular signalling, angiogenesis and embryonic blood-vessel development. Tissue factor can be found both as free (soluble tissue factor) and membrane bound forms. Several studies have shown that measurements of any of these forms may provide clinically significant information, particularly in patients with malignant and inflammatory diseases, and are cost-effective. PMID- 11938387 TI - Physical activity, fitness and fatness among Saudi children and adolescents: implications for cardiovascular health. AB - During recent years, the Kingdom of Saudi Arabia has witnessed a tremendous development at an astounding rate. The standard of living rises and mechanization has been apparent in all aspects of people's life. As industrialization and modernization progress, a number of changes in physical activity and eating habits are likely to occur. Indeed, physical inactivity and sedentary living with associated low level of physical fitness are increasingly becoming prevalent in the Saudi society. These lifestyle changes undoubtedly carry unfavorable consequences on health outcomes of the Saudi population. This paper reviews the status of physical activity among Saudi children and adolescents and discusses its implications to cardiovascular health and fitness. From the available evidences, it appears that most Saudi children and adolescents do not meet the minimal weekly requirement of moderate to vigorous physical activity necessary for effectively functioning cardiorespiratory system. Furthermore, active Saudi boys tend to have favorable levels of serum triglycerides and high density lipoproteins-cholesterol compared with inactive boys. Sixteen percent of Saudi schoolboys are considered obese (fat content is above 25% of body mass). Body fat percent of Saudi boys seems to have increased over the past decade. Body fatness correlated significantly with several coronary artery disease risk factors. Based on the available evidences, promotion of physical activity among Saudi children and adolescents appears warranted and national policy encouraging active living is also needed. PMID- 11938388 TI - Experience with the objective structured examination as a tool for students' assessment in the Department of Community Medicine and Primary Health Care in a university [corrected] in Western Saudi Arabia. AB - OBJECTIVE: The objectives of this study were to 1. assess the knowledge and attitude of the 5th year female medical students towards the objective structured examination in general and in Community Medicine in particular prior to the exam, 2. Detect any change in students' knowledge and attitude after passing the exam, 3. Identify the students' evaluation for future objective structured examination in Community Medicine and Primary Health Care and 4. Compare the students' performance in the objective structured examination in Community Medicine and Primary Health Care to the results of multiple choice questions evaluation in the department. METHODS: There were 2 data sets, the first was collected by self administered pre and post-objective structured examination questionnaires from all female medical students in the 5th year during the academic year 1996-1997 and the 2nd was copied from the students' results for objective structured examination and multiple choice questions in the department of Community Medicine and Primary Health Care during same academic year. RESULTS: All the 34 5th year female medical students were enrolled in the study. Most of them reported that they did not receive enough information prior to the objective structured examination and most of their knowledge came from side chats with friends or staff members which was confirmed by their weak knowledge and negative attitude towards the objective structured examination prior to the exam. After setting the objective structured examination, students' knowledge and attitude towards objective structured examination markedly improved and the majority displayed its advantage as an assessment tool and were convinced of its suitability for medical practice in general and community medicine in particular. Most students believed that objective structured examination in Community Medicine and Primary Health Care met their expectations and recommended it for the coming years. Also, students stated the negative and positive aspects that should be considered for future assessments. Moreover, the multiple choice questions were nearly similar and there was a significant correlation between both scores. CONCLUSION: The 5th year female medical students did not receive enough information and training on the objective structured examination which influenced their attitude prior to the exam. However, most of them showed a positive attitude after the experience and their marks in the objective structured examination were comparable to their marks in the multiple choice questions. The objective structured examination is a popular method of assessment in medical practice worldwide. The decision to discontinue its use after only one year did not allow space of time for evaluation of its appropriateness as an assessment tool. More research is recommended to study the main causes that lead to disregarding this type of medical assessment in the College of Medicine at King Abdulaziz University. PMID- 11938389 TI - Management of oral cancer in a tertiary care hospital. AB - OBJECTIVE: Oral cancer is the 3rd most common cause of malignancy in Saudi Arabia after lymphoma and leukemia. The high incidence of oral cancer in this region is attributed to frequent use of tobacco (Shamma). In this report we have reviewed the mode of treatment and subsequent outcome of 228 oral cancer patients at the Oral Maxillofacial Surgery Unit of the Armed Forces Hospital, Riyadh, Kingdom of Saudi Arabia. METHODS: In this retrospective study the case notes of 228 oral cancer patients treated at the Armed Forces Hospital, Riyadh were reviewed. Age, sex, presenting symptoms, location of cancer in oral cavity, habit of tobacco chewing, mode of treament and complications as a result of treatment were recorded. RESULTS: Out of 228 patients 149 were males and 79 were females with age ranging from 23 to 82 years. The majority of patients (58%) had tobacco chewing habit. Most of the patients received radiation therapy (98%), some times with adjuvant chemotherapy. Seventy-three percent of patients underwent surgery followed by radiotherapy, whereas 2% had surgery alone. Recurrence of the disease was observed in 56% of the patients over a period of 5 years following initial treatment. CONCLUSION: A close association between tobacco chewing and oral cancer was observed. It is concluded that post surgery radiotherapy is a highly successful method of treatment in patients with oral cancer. Cases with a history of shamma usage did not show deep invasion even when the tumor was large. They also showed good prognosis after excision but more resistant to radiotherapy comparing with those cases with no history of shamma usage. This point needs further investigation. PMID- 11938390 TI - Clinicopathological trends in colorectal cancer in a tertiary care hospital. AB - OBJECTIVE: This study aims to retrospectively analyze the pattern of patients with colorectal cancer seen in King Abdul Aziz Hospital and Oncology Center, Jeddah, Kingdom of Saudi Arabia from January 1992 through to December 1997, with a view to determine various epidemiological and clinopathological features of the disease. METHODS: All cases of colorectal cancer presented to King Abdul Aziz Hospital, Jeddah between January 1992 and December 1997 were retrospectively reviewed and the data was analysed to determine age, gender, ethnicity, subsite distribution, clinical presentation, histological type and staging of disease. RESULTS: A total of 160 patients were included in the study, with 78% of them being Saudi Nationals. The male to female ratio was 1:0.8. The mean age was 56.3 + 14.98 years. The peak age of onset was in the 6th and 7th decade. The duration of symptoms varied from one to 24 months with the majority of patients having symptoms for more than 6 months. Sixty-eight point two percent of these patients had primary disease originating from rectum or sigmoid colon, whereas 22.5% of patients had primary disease involving ascending and transverse colon, Anemia was present in 55% of patients. Among the 160 patients, 82.5% of them were histologicaly found to have adenocarcinoma. A total of 38.8% of patients were in stage B and 38.1% of patients were in stage C of Aster-Coller classification. No patients was found to be in stage A. CONCLUSION: In general, we share many epidemiological features of developing countries for colorectal carcinoma. These included left sided subsite distribution and delayed presentation of the disease in an advanced stage. We stress the significance of public health education and a national screening program regarding colorectal cancer to improve the outcome. PMID- 11938391 TI - The diagnosis of gastroesophageal reflux disease in children. AB - OBJECTIVE: Gastroesophageal reflux disease is a common disorder affecting children worldwide. The objective of this study is to report our experience on the accuracy of tests used for the diagnosis ofgastroesophageal reflux disease with emphasis on the advantages and disadvantages of each of them. METHODS: This study took place in the Pediatric Gastroenterology Division, Department of Pediatrics, College of Medicine and King Khalid University Hospital, Riyadh, Kingdom of Saudi Arabia, during the period of 1994 through to 1999. Results of barium meal, 24-hour esophageal pH monitoring, endoscopy, and gastrointestinal scintigraphy are analyzed and compared in children with and without gastroesophageal reflux disease. RESULTS: One hundred and forty-four children were investigated. The diagnosis was confirmed in 85 and excluded in 59 children, who will be considered as patients without gastroesophageal reflux disease. The results of barium meal, 24 hour pH monitoring, endoscopy, and gastrointestinal scintigraphy were positive in 80%, 78%, 92%, and 70% of the patients with gastroesophageal disease. The same studies were falsely positive in 29%, 9%, 19%, and 0% of those without gastroesophageal reflux disease. Esophageal pH was the most specific diagnostic study (91%), whereas endoscopy was the most sensitive (92%) and had the best positive predictive value (95%). CONCLUSION: The results of this study are similar to reports from other parts of the world. It is stressed that all procedures have important advantages and disadvantages indicating that the selection of procedures should be individualized and based on the clinical situation. PMID- 11938392 TI - Risk factors for duodenal ulcer disease. AB - OBJECTIVE: To evaluate the role played by some of the traditional risk factors on the prevalence of duodenal ulcer disease in Jordan, where the prevalence of Helicobacter pylori is high. METHODS: This study was carried out in the Endoscopy unit at the Health Center, Jordan University of Science and Technology, Irbid, Jordan, during the period January 1998 through to july 1999. The patients' group consisted of 290 subjects with endoscopically proven duodenal ulcers and 161 subjects who were endoscoped for various indications and found to have no ulcers were considered as controls. Validated questionnaires were filled out before the procedure, containing data related to age, sex, area of residence, family history of ulcer, blood grouping, cigarette smoking, coffee and tea ingestion, and regularity of meals. RESULTS: The following factors were associated with increased ulcer prevalence, male gender, family history of ulcer, blood group O, skipping breakfast or more than one meal, coffee ingestion and cigarette smoking. CONCLUSION: Many factors are important in the predisposition for ulcer disease in subjects with Helicobacter pylori. Modification of these factors would be essential to decrease the prevalence of ulcer disease and will result in large economic and medical savings. PMID- 11938393 TI - Precipitating factors for diabetic ketoacidosis. AB - OBJECTIVE: The aim of this study is to identify the precipitating factors from a medical and social point of view, in addition to discussing some clinical and laboratory aspects of diabetic ketoacidosis. METHODS: Sixty-eight patients were admitted to King Abdulaziz University Hospital, Jeddah, Kingdom of Saudi Arabia, over a 2 year period, (April 1999 through to April 2001). Diagnosis of diabetic ketoacidosis was based on: clinical features, serum sugar >12 mmol/L with ketonuria, bicarbonate and base deficit. RESULTS: The mean age was 22.5 years (0.5-87) years with a male to female ratio of 1.4:1. Poor compliance to continue the treatment and infection were the most common precipitating factors being responsible for 54.4% and 28% cases. A low mortality rate of 2.9% in our study compared favorably with other studies, which contributed to a high level of medical care in King Abdulaziz University Hospital. CONCLUSION: Diabetic ketoacidosis is a fatal complication among our diabetic patients. Implementing a patient education program to increase awareness of the disease is the most important step in the prevention of this complication. The authorities should ensure availability of insulin to all patients, either free or at lower prices. The role of cultured and socioeconomic factors in aggravating or precipitating diabetic ketoacidosis should always be considered and where possible, eliminated PMID- 11938394 TI - Audit of referral of diabetic patients. AB - OBJECTIVE: To determine the referral, the feedback rates and to find out the prevalence of diabetic retinopathy among diabetics who were referred from a primary health care center, to Abha General Hospital in Aseer Region. METHODS: This study was conducted at Wasat Abha Primary Health Care Center in Aseer region at the end of 1996 and 1997. All the diabetic files were reviewed and evaluated for referral to the eye clinic, their feedback and the prevalence of diabetic retinopathy among diabetics for whom fundoscopy was carried out. The related data was entered and analyzed by using the statistical package for social sciences program. RESULTS: A total of 203 diabetic files were reviewed. Forty percent were referred during 1996 while 68.5% were referred during 1997. The rates of feedback were 72% in 1996 and 71.2% in 1997. Twenty percent were referred twice during these 2 years. The prevalence of diabetic retinopathy was 11.3%. CONCLUSION: The referral rate and the feedback from the eye clinic was lower than the national target. A poor referral system was responsible for such a low rate. Either reorganization of the referral system or calling the ophthalmologists to attend Primary Health Care Centers monthly will increase the rate of eye examination among diabetics and will help in early detection of diabetic retinopathy and in prevention of blindness. PMID- 11938396 TI - Detection of extended-spectrum b-lactamases in members of the family enterobacteriaceae at a teaching hospital, Riyadh, Kingdom of Saudi Arabia. AB - OBJECTIVE: To determine the prevalence of extended spectrum b-lactamase at King Khalid University Hospital, Riyadh, Kingdom of Saudi Arabia and to compare the ability of the disc diffusion, double disc potentiation methods and the extended spectrum b-lactamase E test to detect exended-spectrum b-lactamase among enterobacteriaceae. METHODS: This study was undertaken during the time period period January 1 through to September 30 1999, at King Khalid University Hospital, Riyadh, Kingdom of Saudi Arabia. A total of 187 multiresistant isolates of enterobacteriacae from different clinical specimens were tested for the extended-spectrum b-lactamases. The performance of disc diffusion, double disc potentiation methods using cefotaxime, ceftazidime, ceftriaxone and aztreonamand the extended-spectrum b-lactamase E based on the reduction of the minimum inhibitory concentration of ceftazidime in the presence of clavulanic acid were compared for the detection of extended-spectrum b-lactamase production. RESULTS: Thirty six percent of our isolates produced extended-spectrum b-lactamases. Among these 42% were Klebsiella pneumoniae and 20% were Escherichia coli. Other species of Enterobacteriaceae produced extended-spectrum b-lactamase in low numbers. Disc diffusion method was not suitable for detecting extended-spectrum b-lactamases among the isolates. Double disc potentiation performed comparably well with the extended-spectrum b-lactamases E test. CONCLUSION: Extended-spectrum b-lactamases occurs in Klebsiella pneumoniae and Escherichia coli at a significant number. The use of double disc potentiation method for screening is practical and the extended-spectrum b-lactamase E test with ceftazidime is a useful confirmatory test for extended-spectrum b-lactamase production. PMID- 11938395 TI - Nutritional rickets and osteomalacia in school children and adolescents. AB - OBJECTIVE: To review experiences of nutritional rickets and osteomalacia in school children and adolescents at King Khalid University Hospital, Riyadh, Kingdom of Saudi Arabia. METHODS: Records of children and adolescents aged 6-18 years, seen at King Khalid University Hospital, Riyadh, Kingdom of Saudi Arabia, during the period January 1994 through to December 1999, who were diagnosed to have rickets or osteomalacia were reviewed. The diagnosis was based on clinical, biochemical and radiological data. Data extracted and analyzed included age, sex, presenting symptoms and signs, dietary history and sun exposure, blood count, bone profiles, renal and liver profile, and 25-hydroxy vitamin D3 and 1, 25 dihydroxy vitamin D3. Hand and wrist x-rays were carried out for all patients while bone density of lumbar spine and 3 femoral sites and bone scan were performed on the majority of patients. RESULTS: Forty-two children and adolescents (25 females and 17 males) were diagnosed. Their age ranged between 6 18 years with a mean of 13.5. Non specific symptoms, such as bone pain and fatigue were the most presenting symptoms, while skeletal deformities and fractures were the presenting symptoms in only 5 and 3 patients. Lack of direct sun exposure and poor calcium intake was evident. Bone profiles at the time of diagnosis revealed mean serum calcium of 2.1 mmol/L, range 1.5 2.3 (Normal=2.2 2.7), phosphorus 1.1 mmol/L, range 0.7 1.9 (Normal=1.4 2.1) and alkaline phosphatase activities of 1,480 U/L, range 834-2,590 (N=<600). Serum concentrations of 25-hydroxy Vitamin D were low (<10 mg/L) while that of 1, 25 Dihydroxy Vitamin D varied between low to normal (<10-45 ng/L). Bone density of the lumbar spine and 3 femoral sites were performed in 26 patients and showed markedly reduced values, while bone scan demonstrated a high uptake of tracer throughout the skeleton "super scan". Multiple stress fractures were evident in 8 children. CONCLUSION: Although a community-based study to assess the magnitude of the problem is needed, it seems that rickets and osteomalacia of nutritional origin are not that uncommon and deserves special attention from all pediatricians and practicing physicians. They also suggested that further studies are needed to help understand the pathophysiology, and identify the contributing factors for the development of the disorder. PMID- 11938397 TI - Bone marrow examination. Indications and diagnostic value. AB - OBJECTIVE: To identify the main indications for bone marrow examination in a University hospital setup and the most common diagnoses encountered. To also identify the extent of correlation, if any, between the preliminary diagnosis and the result of the final bone marrow diagnosis. METHODS: The requests and reports of all bone marrow biopsies and aspirations carried out during a 12-year period from January 1988 through to December 1999, in King Fahd Hospital of the University, Al Khobar, Kingdom of Saudi Arabia were retrospectively reviewed. The information extracted included the main indications for performing this procedure, age groups involved, and the most common diagnoses encountered. A specially designed form was used for this purpose and the data was analyzed using the statistical package for social sciences. Randomly selected slides of the most common diagnoses were reviewed to concur with the diagnosis. RESULTS: There was a total of 1813 bone marrow biopsies or aspirations, or both, performed. The main indications for bone marrow examination in a descending order of frequency were the following: The diagnosis and management of acute leukemia 403 (22.2%), staging for lymphoma 276 (15.2%), evaluation of pancytopenia 215 (11.9%), thrombocytopenia 173 (9.5%), investigation of anemia 151 (8.3%), fever (pyrexia of unknown origin) 130 (7.2%), lymphadenopathy 120 (6.6%), and hepatosplenomegaly 80 (4.4%). The most common diagnoses encountered were: acute lymphoblastic leukemia 242 (13.3%), immune thrombocytopenia 123 (6.8%), acute myeloblastic leukemia 80 (4.4%), hypersplenism 79 (4.4%), chronic granulocytic leukemia 73 (4.0%), megaloblastic anemia 66 (3.6%), bone marrow positive for lymphomatous infiltration 63 (3.5%), chronic lymphocytic leukemia 40 (2.2%), and multiple myeloma 32 (1.8%). CONCLUSION: This study confirms that bone marrow examination is a very important investigation for establishing the diagnosis in many conditions, especially hematological neoplasms. The most common indication for this procedure in our hospital was to confirm and manage acute leukaemia and the most common diagnosis was acute lymphoblastic leukemia. PMID- 11938398 TI - Features and complications of nephroptosis causing the loin pain and hematuria syndrome. A preliminary report. AB - OBJECTIVE: Loin pain with and without hematuria is a serious clinical problem that is most difficult to diagnose and treat. The underlying symptomatic nephroptosis was disparaged long ago, the loin pain hematuria syndrome is doubted and the link between the 2 conditions remains unknown. The lack of demonstrable pathology on all supine imaging has caused disbelieve. This article aims to demonstrate underlying symptomatic nephroptosis features and complications including loin pain hematuria syndrome and discuss the patho-etiology mechanism. METHODS: Observational study and thorough investigations of patients presenting with loin pain and hematuria showed that all supine standard and ancillary imaging was normal. Upright intravenous urography and isotope renography, however, showed features of symptomatic nephroptosis causing the initial intermittent renal pain. Retrograde pyelography demonstrated late organic complications of symptomatic nephroptosis causing pain and hematuria of loin pain hematuria syndrome. Observations detected the illusive overlooked anomalies demonstrable on photographs. RESULTS: All supine standard and ancillary imaging appeared deceptively normal in patients presenting with recurrent episodes of loin pain with or without hematuria. Upright imaging demonstrated gross nephroptosis with pelvi-ureteric junction kink, causing the initial intermittent and later organic obstruction pain. Renal pedicle stretch or twist was also demonstrable on upright intravenous urography and isotope renography as causes of ischemic renal pain. Retrograde pyelography demonstrated the organic renal damage of symptomatic nephroptosis when complicated into loin pain hematuria syndrome. Pyelocalyctaisis with eroded papillae, peritubular backflow and intrarenal extravasation of contrast medium with venous leakage, showed the renal site and cause of hematuria fulfilling the definition of loin pain hematuria syndrome. "Auto-nephropexy" and "sympathetic nephroplegia" were illusive neuro-ischemic findings that took years of follow up observation to affirm in cases of symptomtic nephroptosis complicated into loin pain hematuria syndrome. Other complications included segmental infarction and renal atrophy "auto-nephrectomy". CONCLUSION: The presented photographs demonstrate that loin pain and hematuria have real heterogeneous patho-etiology of ureteral kink obstruction and pedicle stretch or twist ischemia of symptomatic nephroptosis with intermittent and irreversible stages. The overlooked anomalies on all supine imaging are demonstrable on upright imaging and retrograde pyelography with pyelocalyctaisis that may affect both kidneys via sympathetic neuropathy. The reproducible evidence affirms that pain is genuine and symptomatic nephroptosis may be complicated into loin pain hematuria syndrome. PMID- 11938399 TI - Psoriasis in the eastern Saudi Arabia. AB - OBJECTIVE: To determine the percentage occurrence, demographic and clinical features of psoriasis in adult patients at King Fahd Hospital of the University, Al-Khobar, Kingdom of Saudi Arabia. METHODS: The medical records of all the adult patients with psoriasis were analyzed from January 1991 to December 1994 at the dermatology clinic, King Fahd Hospital of the University, Al-Khobar, Kingdom of Saudi Arabia. RESULTS: A total of 263 adult psoriatic patients were studied with the percentage occurrence of 5.3%. There was a male preponderance with sex ratio of 1.4:1. The mean age of onset in males was 26.9 years while in females it was 22.3 years. Fifty-three percent of psoriatic cases developed before the age of 30 years. Family history of psoriasis was recorded in 8.4% of the cases. Itching was the only symptom reported by patients in 43% of cases. The sites of involvement were as follows: lower extremity 44.9%, scalp 41.8%, nail 26.6% and palmoplanter 12.6%. Plaque psoriasis was the most common clinical type (87.1%), followed by erythrodermic (4.2%), pustular (3%), guttate (1.9%), flexural, (2.3%) and follicular (0.4%). CONCLUSION: The present study represents the apparent magnitude of psoriasis in the eastern province of Saudi Arabia, which may reflect that the disease is common in our area as elsewhere. The clinical features of psoriasis in our patients were similar to those reported from other parts of the world. PMID- 11938400 TI - Zinc in normal and pathological human prostate gland. AB - OBJECTIVE: This study is conducted to detect quantitatively zinc in the nuclei and cytoplasm of epithelial cells of the prostate from normal, acute prostatitis, benign prostatic hyperplasia and adenocarcinoma. METHODS: Prostatic tissues from normal, acute prostatitis, benign prostatic hyperplasia and adenocarcinoma were obtained from patients and processed for zinc detection using x-ray microanalysis technique. The samples were collected over a period of 2-3 years and were processed at Jordan University of Science and Technology, Irbid, Jordan. RESULTS: Zinc was increased and decreased both in the nuclei and the cytoplasm of the glandular epithelium of benign prostatic hyperplasia and adenocarcinoma of the prostate. Although zinc was increased in the nuclei and cytoplasm of epithelial cells of acute prostatitis, it was not significant. CONCLUSION: These findings might be caused by factors affecting the zinc metabolic pathway directly or through the zinc bound protein metallothionein. In addition, these findings could be used in diagnosing different prostatic pathological conditions and advancing prostatic tumors and those with similar histopathological profiles. PMID- 11938401 TI - Toxic effect of tannic and related compounds on human plasma proteins. AB - OBJECTIVE: To investigate the toxicity of tannic acid related compounds such as gallic acid and polyphenol on the activity of plasma proteins in vitro. Their electrophoretic results show extremely important information, albumin and globulin levels are remarkably changed and characterized by disorder in their fractions avere which occurred frequently. METHODS: All plasma proteins samples of sets A, B and C were treated in sequences with known different concentrations of gallic acid, gallotannin and polypholes, which were separated chromatographically from phenolic extract of fruit peel of punicaceae. These were then treated, A, B and C were subjected to electrophoresis techniques, for identification and quantitation. RESULTS: The electrophoretic patterns of treated plasma proteins samples, sets A, B and C are arised with remarkable changes in their fraction levels, compared to normal. The results in were also characterized by disorders in their electrophoretic pattern. In this way 5 fractions of treated plasma proteins could be distinguished after sustaining which are ablumin and alpha1, alpha2 and beta and gamma globulins. CONCLUSION: The biological activity of tannic acid related compounds on plasma proteins in vitro, is important in determining their toxicity, and this toxicity may be depend upon their metabolic processes in the liver. In addition, the electrophoretic techniques used for separation and identification of plasma protein is extremely important for future work in the area. PMID- 11938402 TI - Cerebellar abscess due to Listeria monocytogenes. AB - Brain abscess due to listeria monocytogenes mainly involves the cerebral hemispheres. Cerebellar abscess is an infrequent event, which could lead to rapid neurological deterioration if unrecognized. We present a case of multiple brain stem and cerebellar abscesses in a previously healthy individual exposed to unpasteurized milk. This is the 2nd case of cerebellar abscess due to listeria monocytogenes reported in the English literature. The diagnosis of listeria monocytogenes was made in the surgical specimen. Our case illustrates the difficulty of early diagnosis of cerebellar listeria monocytogenes abscesses and the importance of prompt neurosurgical intervention. PMID- 11938403 TI - Staphylococcal toxic shock syndrome in a small infant. AB - Non menstrual staphylococcal toxic shock syndrome is rare in small infants. This is a 4-month-old infant presented to us with a picture of bronchiolitis and few postuler skin lesions, treated with antistapylococcal antibiotics in addition to other supportive medications. On the 4th day of therapy the patient developed sunburn like erythroderma, hypotension, and high grade fever. The dose of antibiotics was increased to the maximum possible dose, in addition to other supportive medications. The patient improved and developed extensive desquamation in both hands and feet on the 14th day of hospitalization, which confirms the diagnosis. PMID- 11938404 TI - Myofibroblastoma of the breast in an adolescent. AB - A case of myofibroblastoma of the breast was recently diagnosed in an adolescent Omani boy. In this report, the clinical, histopathological and the radiological features of this rare disease are being discussed. We conclude that, myofibroblastoma of the breast, which was earlier described, predominantly, in elderlies, can occur in people from any age group. PMID- 11938405 TI - Difficulties in diagnosing neuroleptic malignant syndrome. AB - This report describes a recent case of neuroleptic malignant syndrome, a rare adverse effect of antipsychotic medications. Assessment of the neuroleptic malignant syndrome was carried out by using the Naranjo Algorithm Probability Scale which resulted in a highly probable reaction to haloperidol. The difficulties in establishing the diagnosis in this patient lead to late intervention resulting in a slow recovery and prolonged hospitalization. Our objective in this presentation is to highlight the importance of early recognition of this potentially fatal adverse drug reaction. PMID- 11938406 TI - Delayed presentation of bilateral Morgagni's hernia in a child with Down's Syndrome. AB - This case report describes the delayed presentation of bilateral Morgagni's hernia in a 13-month-old girl with Down's Syndrome. The report emphasizes the fact that a previously normal chest x-ray should not preclude the diagnosis of Morgagni's hernia even when bilateral. The various presentations and the association between Morgagni's hernia and Down's Syndrome are also discussed. PMID- 11938407 TI - Post tonsillectomy quinsy. PMID- 11938408 TI - Doctors response to computer virus. PMID- 11938409 TI - Infundibular buckling as an adjunctive sign of pituitary adenoma. PMID- 11938410 TI - Serum ferritin and other iron parameters in patients with pulmonary tuberculosis. PMID- 11938411 TI - High occurrence of diabetic ketoacidosis among type 2 African patients. PMID- 11938412 TI - Meningococcal disease. AB - Meningococcal disease occurs as both endemic and epidemic disease in most parts of the world with significant morbidity and mortality. Among the different serogroups of Neisseria meningitidis, serogroups A, B, C account for 90% of the disease. In the last few years there has been a change in the epidemiology of the disease with an increase in the prevalence of serogroup C in Europe and North America, serogroup Y in the United States of America and Sweden, and W135 in the Kingdom of Saudi Arabia. The emergence of Neisseria meningitidis serogroup W135 in the Kingdom of Saudi Arabia has lead to 2 major outbreaks mainly among Pilgrims during the Hajj season of 2000 and 2001. This has lead the health officials in the Kingdom of Saudi Arabia to change their vaccine requirements for the Umra and Hajj to include the quadrivalent meningococcal vaccine (A, C, Y, W135) instead of the bivalent one (A, C). Despite all the advances in prevention, diagnosis and treatment, the disease continues to have high mortality (5-10%). Prompt empirical treatment for suspected cases should include penicillin or a 3rd generation cephalosporin. A new conjugate vaccine against Neisseria meningitidis serogroup C has been recently licensed, while quadrivalent conjugate vaccine against serogroup A, C, Y and W135 is in early development. Meanwhile targeted vaccination with the available vaccines according to the epidemiology of the disease and rapid chemoprophylaxis for the close contacts of active cases are the most effective preventive strategies. PMID- 11938413 TI - Innovations in pancreas transplantation. AB - Pancreas transplantation is currently the curative treatment for type 1 diabetes mellitus. It aims at providing physiological insulin replacement therapy in type 1 diabetes mellitus. The goal is thereby also to prevent secondary complications of diabetes. Long term control of glucose metabolism has only been achieved by pancreas transplantation. As a result of improvements in the surgical techniques and the efficacy of immunosuppression, the patient and graft survival rates have improved dramatically over the last 2 decades. As a result, pancreas transplantation, as part of simultaneous pancreas and kidney transplantation, pancreas after kidney transplantation, and exceptionally pancreas transplantation alone, became the standard therapeutic option for patients with type 1 diabetes mellitus with end-stage renal disease. In this article we review the pancreas transplantation methods, indications, techniques, and the short as well as the long outcomes of treatment. PMID- 11938414 TI - Vascular anomalies - diagnosis and therapy. AB - OBJECTIVE: Vascular anomalies were once thought to be impossible to properly diagnose and treat. Hence, we aimed to evaluate the different diagnostic and therapeutic modalities in the management of vascular anomalies. METHODS: We carried out a retrospective review of our experience to evaluate different diagnostic and therapeutic modalities in the management of 25 patients with vascular anomalies over a 2-year-period at King Khalid University Hospital, Riyadh, Kingdom of Saudi Arabia and follow-up period ranging from 2 months to 2 years. RESULTS: Vascular anomalies were more common in male patients (N=19). Age range was 7 to 46 years. Vascular anomalies were categorized as hemangioma (N=2) or malformation (N=23). The vascular malformation were further subdivided into slow flow (N=5) and fast flow (N=18). Duplex (N=12) and radiographic studies; angiography (N=21), venography (N=7), computerized tomography (N=10) and magnetic resonnance angiography (N=8) were used to confirm diagnosis. The treatment of hemangiomas were surgical resection (N=1) and conservative treatment (N=1). Embolization was the main modality of treatment in vascular malformation (N=16), with surgical resection in 4 patients, sclerotherapy in one and conservative in the other 2. All cases had successful outcome with no complications. CONCLUSION: Control of large vascular malformations with acceptable results can be achieved nowadays. Intra-arterial embolization is the mainstay of treatment and long term follow-up with serial physical examination, duplex and arteriography is required. PMID- 11938415 TI - Hydroxyurea in sickle cell disease patients from Eastern Saudi Arabia. AB - OBJECTIVE: To assess the efficiency and safety of hydroxyurea in patients with sickle cell disease from the Eastern Province, Kingdom of Saudi Arabia. METHODS: The study was an open-label and uncontrolled trial. Patients older than 10 years of age with sickle cell disease who suffered 4 or more episodes of painful vaso occlusive crises requiring admissions per year were included, 36 patients (23 males and 13 females) were included between June 1994 and June 1998. Patients were started on hydroxyurea at a dose of 8-10mg/kg per day and the dose was escalated to a maximum tolerated dose or a dose of 35 mg/kg per day. Blood count, renal and liver functions, and hemoglobin F levels were monitored regulary. Clinical response was assessed by record of number of vaso-occlusive crises, requirement for hospital admission and self scoring at the end of each year of treatment. RESULTS: Thirty-six patients were enrolled in the study until the time of analysis of the data. The data of the first 27 patients (18 males and 9 females) who completed 12 months of therapy were analyzed and presented. There was significant reduction in leukocyte, platelet counts and rise in total hemoglobin and hemoglobin F. Hemoglobin F rose by 1.2-13 folds, from the baseline. Seventy-four percent of patients had at least 2 fold rise of maximum hemoglobin F. The mean maximum tolerated dose of hydroxyurea was 16.4 mg/kg. There was significant reduction in hospital admissions and hospital stay. No major side effects had occured. CONCLUSION: Hydroxyurea seems to be effective in decreasing the frequency of vasooclusive crises in patient with sickle cell disease from Eastern Saudi Arabia. In this preliminary analysis no major side effects were observed. Long term side effects need to be monitored. PMID- 11938416 TI - Lymphadenopathy in adults. A clinicopathological analysis. AB - OBJECTIVE: To describe the clinicopathological features of patients admitted to adult wards with a primary presenting feature of lymphadenopathy. METHODS: A retrospective study of all patients admitted to Riyadh Medical Complex, Riyadh, Kingdom of Saudi Arabia between April 1996 through to March 2000. RESULTS: The patients mean age was 35.2 years (standard deviation 15.7 years) and 59.5% were females while 56.2% were Saudis. Swelling was the primary presenting symptom in 39.3% of the patients, while fever, night sweats, and pain occurred in 17.3%, 10.3% and 8.5% patients. There was considerable overlap between benign and malignant causes of lymphadenopathy but in 56.6% of patients, lymphadenopathy was the only sign. The cervical group of lymph nodes was the most commonly affected site. Hepatomegaly, splenomegaly and both organ enlargement occurred in 59 (22.9%) patients. The presence of generalized lymphadenopathy and other physical signs (serositis and organomegaly) highly suggested a malignant process. Granulomatous lymphadenopathy due to tuberculosis was the most frequent pattern 98 (37.9%) followed by lymphoma 85 (32.9%). Tuberculosis was also more common in females than males (51% versus 19%). Among patients with Hodgkin's disease, nodular sclerosis was the most frequent (75%). Routine investigations did not discriminate between benign and malignant causes of lymphadenopathy. The overall mortality rate was 8.1%: with metastatic disease accounting for 52.4%. CONCLUSION: There were significant similarities and differences between these findings and other previous studies in the Kingdom of Saudi Arabia and elsewhere. PMID- 11938417 TI - Efficacy of erythropoietin in premature infants. AB - OBJECTIVE: To identify the effect of early parental recombinant human erythropoietin and iron administration on the blood transfusion requirement of premature infants. METHODS: In a controlled clinical trial conducted at the neonatal intensive care unit of Al-Hada Military Hospital, Taif, Kingdom of Saudi Arabia over a 16 months period, we assigned 20 very low birth weight infants with gestational age of (mean +/- standard error of mean) 28.4 +/- 0.5 weeks and birth weight of (mean +/- standard error of mean) 1031 +/- 42 gm, to receive either intravenous recombinant human erythropoietin 200 U/kg/day and iron 1mg/kg/day or conventional therapy over a 21 day study period. Blood transfusion administration undergoes a strict protocol in our nursery. RESULTS: During the 3 week study period, the hemoglobin and hematocrit remained similar in the 2 groups while the reticulocyte counts were greater in the recombinant human erythropoietin recipients on day 14. The number and volume of blood transfusions were similar in both groups. CONCLUSION: Very low birth weight infants receive fewer blood transfusions than the number previously reported. Strict phlebotomy and transfusion criteria could minimize the need for human recombinant erythropoietin. PMID- 11938418 TI - A new focus of cutaneous leishmaniasis caused by Leishmania tropica. AB - OBJECTIVE: To determine the epidemiological status of the cutaneous leishmaniasis outbreak, isolation and identification of the parasite and preparation of a control program in a recently identified focus located in the Province of Yazd, Central Iran. METHODS: This study was conducted in 124 classrooms of primary school children and among 139 households of an infected quarter in the city of Yazd during 1999-2000. All the students and members of the households were questioned and examined for the presence of ulcer(s) or scar(s). For each case a form was completed and necessary information were recorded. Smears were prepared from the edge of the ulcer and examined for the presence of amastigotes. Leishmania stocks from human lesions were isolated. Cultured promastigotes were identified using Random Amplified Polymorphic DNA-Polymerase Chain Reaction. Stray dogs were shot in the city and examined for the presence of any ulcer(s) or scar(s). RESULTS: An endemic focus of cutaneous leishmaniasis caused by Leishmania tropica was detected in Yazd City, Central Iran. Examination of 3176 students aged 6-16 years old in 124 primary schools showed a rate of 0.7% for scars and 0.2% for active lesions. There was no significant differences between the sexes in the prevalence of active lesions or scars. A study of prevalence among 139 households with a total population of 572 persons in Darvazeh Ghassabha quarter showed a prevalence of 13.5% for scars and 0.5% for ulcers. The most highly infected age group was 5-9 with a rate of 1.8%. Males and females were equally infected. Forty-six stray dogs were examined and none of them appeared to be infected. CONCLUSION: According to this study, cutaneous leishmaniasis due to Leishmania tropica is endemic in the city of Yazd, Central Iran. Humans are considered to be the main reservoir and the transmission is believed to be from human to human by Phlebotomus sergenti. This focus now can be added to the list of anthroponotic cutaneous leishmaniasis foci in the country. Any residual insecticide spraying is not recommended during the next year due to the low prevalence of the disease. PMID- 11938419 TI - Human parvovirus B19 infection among patients with chronic blood disorders. AB - OBJECTIVE: In a normal host, parvovirus infection can be asymptomatic or can result in erythema infectiosum or arthropathy. Patients with underlying hematologic and immunologic disorders who become infected with this virus are at risk for aplastic anemia. This small study attempts to confirm this relation between the human parvovirus B19 infection as one of the predisposing factor of aplastic crisis in patients with hemolytic disorders. METHODS: The laboratory records of 73 patients' serum samples, which were tested for detection of specific Immunoglobulin M and Immunoglobulin G antibody by means of the recurrently available indirect enzyme linked Immunoassay during the period from March 1998 to March 2001, were reviewed retrospectively at the Armed Forces Hospital, Riyadh, Kingdom of Saudi Arabia. RESULTS: For all patients there were 11 (15%) who were diagnosed as acute infections while 50 (68%) had serological evidence of previous exposure. Eight out of the 11 acute patients had chronic hemolytic disorder as the underlying disease while, the 3 other patients were organ transplant and connective tissue disease patients. CONCLUSION: Seventy eight percent of our infected patients were known to have an underlying blood disorder, while 22% had immunosuppressed disorders such as organ transplant and connective tissue disorder. Parvovirus B19 can be considered as one of the predisposing factors of hemolytic crisis in patients with chronic hemolytic disease. PMID- 11938420 TI - The prevalence of hepatitis B core antibody positivity in donors for liver transplantation in Saudi Arabia. AB - OBJECTIVE: The risk of transmitting hepatitis B virus from donors who are positive for hepatitis B core antibody but negative for hepatitis B surface antigen has been a major concern in liver transplantation. In the Kingdom of Saudi Arabia the overall prevalence of hepatitis B core antibody among the general population was reported to be very high indeed. The purpose of this study is to establish the prevalence of hepatitis B core antibody positivity among liver donors who are negative for hepatitis B surface antigen and offered for liver transplantation in the Kingdom of Saudi Arabia. METHODS: Hepatitis B serological markers were studied in 145 of 209 donors offered for organ transplantation over a 4-year period in King Fahad National Guard Hospital, Riyadh, Kingdom of Saudi Arabia. RESULTS: Out of 145 donors, 51 donors tested positive for hepatitis B core antibody but negative for hepatitis B surface antigen with an over all prevalence of 35.2%. The majority of donors were non Saudi (75.2%), and predominantly from the Indian subcontinent and Far East. The prevalence of hepatitis B core antibody positive donor was significantly higher in non-Saudi (41.3%) compared with Saudi nationals (16.7%). CONCLUSION: In the view of this high prevalence, we believe that all donors considered for liver transplantation in the Kingdom of Saudi Arabia should be tested for hepatitis B core antibody. Furthermore, an algorithmic approach should be developed to minimize the risk of transmitting hepatitis B virus from donors to liver recipients, at the same time not to affect the existing small pool of available donor organs in the Kingdom of Saudi Arabia. PMID- 11938421 TI - Treatment of patients with chronic hepatitis C with normal liver enzymes. AB - OBJECTIVE: This is a controlled prospective study to evaluate the efficacy of induction combination therapy using alpha-interferon and ribavirin in patients with chronic hepatitis C and persistently normal liver enzymes. METHODS: Forty six patients with compensated liver disease tested positive for hepatitis C virus antibody between October 1998 and August 2000 at King Abdulaziz University Hospital and Al-Badriyah Towers, Jeddah Clinics, Jeddah, Kingdom of Saudi Arabia. Twelve patients had persistently normal liver enzymes for 6 months and 34 patients with abnormal liver enzymes (control). Patients were treated using induction combination therapy. Viral load response was measured using branched deoxyribonucleic acid signal amplification test. RESULTS: Ten patients with normal liver enzymes (Group A) and 30 patients with abnormal liver enzymes (Group B) were included. End of treatment response in Group A was 90% as compared to 60% in group B; sustained virologic response in group A was 40% as compared to 43.3% in Group B. CONCLUSION: Response rate to induction combination therapy with alpha interferon and ribavirin in patients with normal liver enzymes was similar to that in those with increase liver enzymes. PMID- 11938422 TI - Risk factors for drug-resistant Mycobacterium tuberculosis in Saudi Arabia. AB - OBJECTIVE: To identify rates of primary and secondary drug-resistant Mycobacterium tuberculosis and their risk factors from a tertiary-care center in the Kingdom of Saudi Arabia. METHODS: Review of microbiological and clinical data of all patients with positive isolates of Mycobacterium tuberculosis between 1995 and 2000 at King Faisal Specialist Hospital and Research Centre, Riyadh, Kingdom of Saudi Arabia. RESULTS: Susceptibility to antituberculosis agents was tested in 320 isolates from 320 patients. The median age was 50 years. Pulmonary tuberculosis was diagnosed in 106 (33%) patients, extrapulmonary in 183 (57%), and both in 31 (10%) patients. Two hundred forty-six isolates were sensitive to all 5 first line agents. Resistance to at least one of the first line agents was documented in 36 (11.3%) isolates. For the year 2000, resistance rates increased to 17.6%. Monoresistance was noted in 20 isolates (6.3%) and polyresistance in 16 isolates (5.0%) including 9 multidrug-resistant Mycobacterium tuberculosis isolates (2.8%). Resistance rates for antituberculosis agents are: Isoniazid, 9.1%; Rifampin, 2.8%; Ethambutol, 1.6%; Streptomycin, 5%; Pyrazinamide, 3.6%. Seventy-eight percent of the resistant isolates are considered primary resistance. History of antituberculosis therapy was the only risk factor associated with drug resistant Mycobacterium tuberculosis, odds ratio 19.9 (P< 0.00001). The mean age of patients with resistant isolates was 42 years compared to 49 years in patients with susceptible isolates (P= 0.047). CONCLUSION: In a population of mostly Saudi patients, primary and secondary drug-resistant Mycobacterium tuberculosis is relatively low but has increased lately. Previous history of antituberculosis chemotherapy and young age are risk factors identified. PMID- 11938423 TI - Hearing impairment in low birth weight children. AB - OBJECTIVE: To investigate the prevalence of hearing impairment in normal and low birth weight children. METHODS: A survey of 9540 Saudi children below 15 years of age was carried out. The survey was performed in the provinces of the Kingdom of Saudi Arabia from September 1994 to May 1998. The subjects were classified into low birth weight (up to 2.5 kg) and normal birth weight (>2.5 kg) groups. Each child was carefully examined for hearing status. RESULTS: The results of this study showed significantly high prevalence of hearing impairment in low birth weight children (14.87%) as compared to normal birth weight children (9.78%). CONCLUSION: Further studies are warranted to identify the potential risk factors associated with the high prevalence of hearing impairment in low birth weight children. PMID- 11938424 TI - Standard penile size for normal full term newborns in the Saudi population. AB - OBJECTIVE: To establish norms for penile size for normal full term Saudi newborns. METHODS: The length between the pubic ramus and the tip of the glans was measured in 379 normal full term Saudi newborns. An unmarked spatula was used where its edge was placed against the pubic ramus and the shaft of the penis was stretched to the point of increased resistance. The penile circumference was measured using a non- stretchable plastic tube placed around the midshaft of the uncircumcised penis. The study was carried out in King Khalid University Hospital and King Fahad National Guard Hospital, Riyadh, Kingdom of Saudi Arabia for the period September 1998 to February 1999. RESULTS: The mean penile length was 3.55 cm (standard deviation 0.57) and the mean penile midshaft circumference was 3.96 cm (standard deviation 0.44 cm). CONCLUSION: The mean penile length of 3.55 cm in this study was similar to previously reported international data. The cut-off lower limit (-2.5 standard deviation) is calculated to be 2.13 cm. The cut-off upper limit (+2.5 standard deviation) is 4.98 cm. The mean penile circumference was 3.96 cm (standard deviation 0.44). PMID- 11938425 TI - Changing trends of histopathology in childhood nephrotic syndrome in western Saudi Arabia. AB - OBJECTIVE: It is widely accepted that minimal change nephrotic syndrome is the most common cause of nephrosis in children. Recent studies have demonstrated an increasing incidence of focal segmental glomerulosclerosis in adults and children. This study was conducted to analyze the trend of histopathologic subtypes in idiopathic nephrotic syndrome in the western area of the Kingdom of Saudi Arabia. METHODS: To determine the possible changes in the etiology of childhood nephrosis, the clinical charts of 46 pediatric patients diagnosed with idiopathic nephrotic syndrome between 1997 and 2001, group A, were reviewed. Results were compared with our initial published data of 132 patients with idiopathic nephrotic syndrome diagnosed between 1983 and 1992, group B. RESULTS: There was no difference in the percentage of biopsies between group A (43%) and group B (31.8%), P= 0.9. There was a decline in the incidence of minimal change nephrotic syndrome in recent years. Even if we assume that all patients without a histological diagnosis had minimal change nephrotic syndrome, presumptive minimal change nephrotic syndrome, the total incidence of minimal change nephrotic syndrome (biopsy proven + presumptive) in group A, was only 65% compared to 79.5% in group B, P=0.02. The incidence of focal segmental glomerulosclerosis was significantly greater in biopsies performed in the recent period, group A, 35% versus group B, 16.7%, P=0.05, and in the total number when we included the presumptive minimal change nephrotic syndrome, group A, 15.2% and group B, 5.3%, P<0.0001. Similarly we found an increased incidence of membranoprolifrative glomerulonephritis in total patients when we included the presumptive minimal change nephrotic syndrome, group A, 13.0% and group B, 5.3%, P=0.02. CONCLUSION: There is a shift toward an increasing prevalence of focal segmental glomerulosclerosis and membranoprolifrative glomerulonephritis over the years in the western area of the Kingdom of Saudi Arabia. These findings may have significant implications in the management of childhood nephrotic syndrome. PMID- 11938426 TI - Colorectal cancers in Saudi Arabia. AB - OBJECTIVE: The aim of this retrospective study is to delineate the histopathological profile of colorectal cancers seen at King AbdulAziz University Hospital, Jeddah, Kingdom of Saudi Arabia and to compare our findings with 12 other studies published in the literature from other areas of the Kingdom of Saudi Arabia. METHODS: Our study group consisted of 39 colorectal cancer patients, who were examined in the department of Histopathology at King AbdulAziz University Hospital, Jeddah, Kingdom of Saudi Arabia, over a period of 4 years from January 1996 to December 2000. The data was retrieved and analyzed. The histopathological characteristics of the tumor such as tumor differentiation, tumor Duke's staging and grade of all the tumors were studied and compared with age of the patients. RESULTS: We received 2552 gastrointestinal endoscopic specimens during a 4 year period; out of these 276 were colorectal specimens. We found 39 cases (21 males and 17 females) of colorectal cancer during this period, which constituted 1.5% of total endoscopic biopsies and 13.8% of colorectal biopsies. Among these cases there were 31 cases (81%, mean age 56) of colorectal adenocarcinoma, 3 cases (7.6%, mean age 35) of signet cell carcinoma, 2 cases (5%, mean age 55.5) of mucinous adenocarcinoma, one case (2.5%, age 68) of metastatic papillary carcinoma, one case of mixed mucin secreting signet cell carcinoma (2.5%, age 64) and one case (2.5%, age 55) of poorly differentiated squamous cell carcinoma. In our study 21% of patients presented in their 3rd decade of life, 18.4% in the 4th decade, 15.8% in the 5th decade, 26.3% in the 6th decade, 10.5% in the 7th decade and 7.9% in the 8th decade. CONCLUSION: Colorectal carcinoma showed frequent presentation in our population. Some malignant lesions showed early presentation as compared to United States of America, with 6.9% cases presenting at a young age and 23.6% of cases presented at middle age. In order to achieve early diagnosis, a comprehensive cancer education program should be planned and executed, and proper screening programs should be launched. PMID- 11938427 TI - Diversity in polyp pathology and distribution of Familial Juvenile Polyposis Syndrome. AB - OBJECTIVE: Juvenile polyposis syndrome is a rare autosomal dominant disorder with incomplete penetrance. The aim of this study was to review our experience with juvenile polyposis syndrome with emphasis on the diversity of polyp pathology and distribution and the recommended treatment. METHODS: Over the period January 1994 through February 2001, 10 family members were managed at Princess Basma Teaching Hospital, Irbid, Jordan. Two siblings with juvenile polyposis syndrome are discussed. RESULTS: The polyps were unusually concentrated in the rectum. In one patient the polyps were purely of the adenomatous type. The father suffered from non-polyposis colon cancer at the age of 35. CONCLUSION: Proctocolectomy and ileal pouch-anal anastomosis is recommended as the treatment of choice. Screening of juvenile polyposis syndrome patients and their relatives is emphasized for early detection of malignancy. PMID- 11938428 TI - Precocious puberty in a female with Proteus Syndrome. AB - Proteus syndrome is a complex disorder comprising malformations and overgrowth of multiple tissues and characterized by its polymorphism and mosaicism. The syndrome is rare and sporadic. Oliveira M da C et al reported the first case of Proteus syndrome associated with precocious puberty in a boy. We are reporting a case of a 7-month old girl with Proteus syndrome who developed a juvenile granulosa cell tumor in one ovary causing precocious puberty. At our knowledge this is the first case of Proteus syndrome with precocious puberty in a female. PMID- 11938429 TI - Congenital lobar emphysema. AB - Congenital lobar emphysema is a very rare congenital cystic malformation of the lung that can cause acute respiratory distress in early life. This paper reviews 6 cases of congenital lobar emphysema seen over a period of 10 years. The medical records of children with the diagnosis of congenital lobar emphysema were retrospectively reviewed for age at diagnosis, sex, presenting symptoms, investigations, treatment and outcome. There were 4 males and 2 females, and all of them presented before 6 months of age. Three presented with recurrent chest infection, while the other 3 had acute respiratory distress soon after birth. In all, the diagnosis was confirmed by chest x-ray, and the left upper lobe was affected in all of them. Although congenital lobar emphysema is rare, clinical awareness of this condition is important for early diagnosis and effective surgical treatment. PMID- 11938430 TI - An unusual presentation of metabolic cardiomyopathy due to Pompe's disease. AB - An Omani infant boy who presented in the neonatal period with cardiac failure secondary to hypertrophic cardiomyopathy is reported. He subsequently progressed to show features of a metabolic disorder with multisystem involvement and was diagnosed to have Type II glycogenosis (Pompe's disease). The differential diagnosis and management of metabolic cardiomyopathy are outlined. PMID- 11938431 TI - Non operative management of blunt bile duct injuries in children. AB - Bile duct injuries after blunt abdominal trauma are rare in children. This report describes 2 cases of blunt bile duct injuries in children, which were managed non operatively. One patient sustained partial tear in the right hepatic duct, which was managed successfully by passing a stent into the injured duct during endoscopic retrograde cholangio pancreatography. The 2nd patient had intrahepatic bile duct injury associated with liver trauma and was managed with a drain placed in the sub-hepatic area percutaneously. This non-operative management in our patients allowed healing of bile duct injuries without any stricture, at 2 years follow-up. Both children are symptom free with normal liver function tests and normal liver ultrasound. With the advancement in radiological and endoscopic techniques, some of the bile duct injuries can be managed conservatively. Conservative treatment must be individualized according to the site and extent of injury. PMID- 11938433 TI - Geriatric care - a challenge to internists in Saudi Arabia. PMID- 11938432 TI - Splenic abscess in childhood B-Thalassemia Major. AB - Splenic abscess is a rare complication of thalassemia major. In this paper we report a 10-year-old male thalassemic child with splenic abscess. He presented with high-grade fever, abdominal pain and tender splenomegaly. The diagnosis was confirmed by ultrasonography and computerized tomography scan of the abdomen. The patient underwent emergency splenectomy. PMID- 11938434 TI - Spontaneous rupture of an ovarian tumor during labor. PMID- 11938435 TI - Etiology of pleural effusion in Western Saudi Arabia. PMID- 11938436 TI - Acute appendicitis presenting as urinary retention: a note of caution. PMID- 11938437 TI - Merosin-deficient congenital muscular dystrophy, autosomal recessive (MDC1A, MIM#156225, LAMA2 gene coding for alpha2 chain of laminin). AB - Congenital muscular dystrophies (CMDs) are a highly heterogeneous group of neuromuscular disorders. A subgroup displays a specific deficiency in a protein of the extracellular matrix, the alpha2 chain of laminin-2 (merosin). A number of mutations in the gene encoding this protein have been identified in patients who present with a severe phenotype and white matter changes. PMID- 11938438 TI - Second family with hearing impairment linked to 19q13 and refined DFNA4 localisation. AB - Until now, over 30 loci have been identified by linkage analysis of affected families that segregate non-syndromic and dominantly inherited forms of hearing impairment (DFNA). A German family with a non-syndromic progressive hearing impairment transmitted in autosomal dominant mode was linked to 19q13.3-q13.4 by a genome-wide scan. Due to the low lod-score (1.85 at theta=0.05) for APOC2-locus we extended the fine mapping attempt with further markers in the same chromosomal region. This resulted in significant evidence for linkage to the markers D19S246 and D19S553 (two-point lod-score of 4.05 and 3.55 at theta=0.0) and a candidate critical region of 14 cM between markers D19S412 and D19S571. This region shows partial overlap with the previously reported DFNA4 critical region. The human gene BAX is orthologous to the rodent Bcl2-related apoptosis gene that is temporally expressed during the postnatal period in the developing inner ear of the mouse. BAX, mapping at a distance of no more than 0.73 cM distally to marker D19S553 appeared a likely candidate in our pedigree but genomic sequencing of coding regions and exon/intron boundaries excluded disease-related mutations. However, additional ESTs in the same region remain to be tested. PMID- 11938439 TI - Determination of the relative contribution of three genes-the cystic fibrosis transmembrane conductance regulator gene, the cationic trypsinogen gene, and the pancreatic secretory trypsin inhibitor gene-to the etiology of idiopathic chronic pancreatitis. AB - In the last 5 years, mutations in three genes, the cystic fibrosis transmembrane conductance regulator (CFTR) gene, the cationic trypsinogen (PRSS1) gene, and the pancreatic secretory trypsin inhibitor (PSTI) gene, have been found to be associated with chronic pancreatitis (CP). In this study, using established mutation screening methods, we systematically analysed the entire coding sequences and all exon/intron junctions of the three genes in 39 patients with idiopathic CP (ICP), with a view to evaluating the relative contribution of each gene to the aetiology of the disease. Our results demonstrate that, firstly, 'gain-of-function' mutations in the PRSS1 gene may occasionally be found in an obvious ICP subject. Secondly, presumably 'loss-of-function' mutations in the PSTI gene appear to be frequent, with a detection rate of at least 10% in ICP and, finally, abnormal CFTR alleles are common: at least 20% of patients carried one of the most common CFTR mutations, and about 10% of patients were compound heterozygotes, having at least one 'mild' allele. Thus, in total, about 30% of ICP patients carried at least one abnormal allele in one of the three genes, and this is the most conservative estimate. Moreover, a trans-heterozygous state with sequence variations in the PSTI/CFTR genes was found in three patients. However, an association between the 5T allele in intron 8 of the CFTR gene and ICP remains unproven. PMID- 11938440 TI - A human interstitial telomere associates in vivo with specific TRF2 and TIN2 proteins. AB - Mammalian telomeres are composed of long arrays of TTAGGG repeats that form a nucleoprotein complex which protects the chromosome ends. Human telomere function is known to require two TTAGGG repeat factors, TRF1 and TRF2, and several interacting proteins, but the mechanism by which the DNA/protein complex prevents end to end fusion in vivo has not been elucidated. In order to better understand the role of specific telomere-associated proteins in the organisation of chromosome ends, we have studied a patient with a rare chromosome rearrangement that has given rise to an interstitial telomere. Using specific antibodies and immuno-FISH on unfixed metaphase chromosomes, we show that the proteins TRF2 and TIN2 (TIN2 interacts with TRF1) co-localise with the interstitial TTAGGG repeats. Our results demonstrate, for the first time in humans, that TRF2 and TIN2 proteins associate with interstitial duplex TTAGGG repeats, in vivo. They confirm that double stranded-telomeric repeats, even when complexed with specific proteins, are not sufficient to create a functional telomere. Finally, they suggest a possible role for proteins in stabilising interstitial TTAGGG repeats. PMID- 11938441 TI - Increased frequency of combined methylenetetrahydrofolate reductase C677T and A1298C mutated alleles in spontaneously aborted embryos. AB - The pathogenesis of spontaneous abortion is complex, presumably involving the interaction of several genetic and environmental factors. The methylenetetrahydrofolate reductase (MTHFR) gene C677T and A1298C polymorphisms are commonly associated with defects in folate dependent homocysteine metabolism and have been implicated as risk factors for recurrent embryo loss in early pregnancy. In the present study we have determined the prevalence of combined MTHFR C677T and A1298C polymorphisms in DNA samples from spontaneously aborted embryos (foetal death between sixth and twentieth week after conception) and adult controls using solid-phase minisequencing technique. There was a significant odds ratio of 14.2 (95% CI 1.78-113) in spontaneously aborted embryos comparing the prevalence of one or more 677T and 1298C alleles vs the wild type combined genotype (677CC/1298AA), indicating that the MTHFR polymorphisms may have a major impact on foetal survival. Combined 677CT/1298CC, 677TT/1298AC or 677TT/1298CC genotypes, which contain three or four mutant alleles, were not detected in any of the groups, suggesting complete linkage disequilibrium between the two polymorphisms. The present finding of high prevalence of mutated MTHFR genotypes in spontaneously aborted embryos emphasises the potential protective role of periconceptional folic acid supplementation. PMID- 11938442 TI - A centenarian-only approach for assessing gene-gene interaction in human longevity. AB - In this study, we introduce a centenarian-only approach to the assessment of gene gene interaction that contributes to human longevity. This approach corresponds to the non-traditional case-only method in the genetic study of gene and disease associations. We first describe how the method can be implemented to screen for gene-gene interaction in human longevity. Then we apply the method to centenarian data collected from an Italian centenarian study in order to detect the interactions between the REN gene and the mitochondrial haplotypes. A significant interaction between REN gene allele 10 and the mitochondrial H haplotype, which may favour longevity, was found. Important features of the application in human longevity studies are highlighted and discussed. Since centenarians constitute a special population representing successful ageing, the centenarian-only approach will be an important tool in the search for major genes that contribute to human longevity. PMID- 11938443 TI - Optimal selection strategies for QTL mapping using pooled DNA samples. AB - The cost of large-scale association studies may be reduced substantially by analysis of pooled DNA from multiple individuals. Here we examine the optimal symmetric and asymmetric designs for pooling experiments for quantitative traits under a range of assumptions about the underlying genetic model and the sources of experimental errors in allele frequency estimation. The results indicate that, in the absence of experimental errors and for common alleles with additive effects, a symmetric pooling scheme comparing the top 27% with the bottom 27% of the trait distribution is optimal, extracting 80% the total information available. A symmetric design is not optimal for rare or recessive alleles, which require asymmetric (or other) pooling strategies. Allele frequency measurement errors reduce the optimal pooling fraction as well as the overall efficiency of the pooling design. In contrast, random variation in the amount of DNA contributed by individuals to a pool reduces only the overall efficiency of the pooling design. Our results emphasize the importance of minimising experimental errors and suggest a pooling fraction of around 20%. PMID- 11938444 TI - An evaluation of the variance components approach: type I error, power and size of the estimated effect. AB - In order to evaluate the statistical properties of the variance components method implemented in SOLAR and GENEHUNTER2, the type I error rate, power and estimated size of modelled effects were determined using computer simulation. Results suggested that the type I error rate was quite conservative when the variance was completely due to random effects. However, when either a polygenic effect or an unlinked single locus component effect was included in the generating models the type I error rate was closer to the nominal level. The size of the polygenic or single locus effect appeared to influence the type I error rate. Results suggested that the variance components method underestimated the variance attributed to a single locus effect and overestimated the variance attributed to a polygenic effect in the models considered regardless of the source of variation. PMID- 11938445 TI - Do reduced levels of steroid 21-hydroxylase confer a survival advantage in fetuses affected by sex chromosome aberrations? AB - We investigated whether molecular defects in the CYP21 gene were detectable in two common sex chromosome aberrations, the Turner and the Klinefelter syndromes. We found abnormal 17-hydroxyprogesterone levels after adrenal stimulation in 26/60 (43.3%) patients affected by these chromosome aberrations, as compared with only 11/68 (16.2%) normal controls (P=0.0014, odds ratio 4.0). Screening of the CYP21 gene identified a single Val281Leu missense mutation in exon 7 in 9/63 (14.3%) of the patients, all nine of whom were heterozygote carriers; the mutation frequency was significantly higher than in the general population (P=0.007, odds ratio=3.5). The hormonal and molecular data indicate that these common sex chromosome aberrations are associated with a remarkably high frequency of steroidogenic defects. It may be hypothesised that reduced levels of steroid 21-hydroxylase could confer a survival advantage, leading to a successful pregnancy. PMID- 11938446 TI - Leigh disease associated with a novel mitochondrial DNA ND5 mutation. AB - Leigh disease is a genetically heterogeneous, neurodegenerative disorder of childhood that is caused by defects of either the nuclear or mitochondrial genome. Here, we report the molecular genetic findings in a patient with neuropathological hallmarks of Leigh disease and complex I deficiency. Direct sequencing of the seven mitochondrial DNA (mtDNA)-encoded complex I (ND) genes revealed a novel missense mutation (T12706C) in the mitochondrial ND5 gene. The mutation is predicted to change an invariant amino acid in a highly conserved transmembrane helix of the mature polypeptide and was heteroplasmic in both skeletal muscle and cultured skin fibroblasts. The association of the T12706C ND5 mutation with a specific biochemical defect involving complex I is highly suggestive of a pathogenic role for this mutation. PMID- 11938447 TI - The differential contribution of MEFV mutant alleles to the clinical profile of familial Mediterranean fever. AB - Familial Mediterranean fever (FMF) is an autosomal recessive disorder characterised by recurring attacks of fever and serositis. Five sequence alterations (M694V, V726A, M680I, M694I and E148Q), in the MEFV gene, account for the majority of FMF chromosomes. The wide clinical variability of the disease has been related to MEFV allelic heterogeneity. M694V homozygotes have a severe form of the disease. Mutations E148Q and V726A have reduced penetrance. The clinical features, associated with the M680I and the complex V726A-E148Q allele, are not well defined. This study aims to further characterise the phenotypic profile associated with the major MEFV mutations. We investigated 220 FMF patients, in whom both FMF alleles have been identified, and found that different genotypes are characterised by a specific allelic related clinical profile and penetrance. Homozygotes for the M694V mutation and the complex V726A-E148Q allele are the most severely affected and often endure renal amyloidosis. Homozygotes for the M680I and V726A alleles and compound heterozygotes for either the M694V or the V726A-E148Q alleles in combination with either the E148Q, the V726A or the M680I alleles are significantly less severely affected. The morbididity associated with the complex V726A-E148Q allele by far outweighs that associated with the V726A allele, bearing evidence to the fact that the E148Q mutation is not a benign polymorphism. These findings increase our understanding of the role of allelic variability in disease expression. PMID- 11938448 TI - BRCA mutations in Italian breast/ovarian cancer families. PMID- 11938449 TI - 4th International Meeting on Single Nucleotide Polymorphism and Complex Genome Analysis. Various uses for DNA variations. AB - At the 4th International Meeting on Single Nucleotide Polymorphism and Complex Genome Analysis (Stockholm, Sweden, 10th-14th October 2001), approximately 100 scientists from more than 20 nations undertook a probing review of latest developments in the field. Despite impressive and still ongoing activities towards SNP discovery and validation, plus efforts towards haplotype exploitation, it was clear that supporting technologies for genotyping are way behind where they need to be. Innate complexity and large variances in aspects of genome function together pose immense challenges that are difficult to surmount in the human situation. In contrast, studies in simpler organisms and population/evolutionary genetics studies are yielding important new insights. Breakthroughs that are being made in understanding the genetic etiology of complex disease tend to involve genes of larger effect or extremely well merited candidates. Linkage studies and proximal phenotypes are being recommended, though the best way forward is still hotly debated. Consequently, many diverse and ambitious projects are underway, from which the data itself will eventually show what is and is not possible. PMID- 11938450 TI - Intrabody-mediated phenotypic knockout of major histocompatibility complex class I expression in human and monkey cell lines and in primary human keratinocytes. AB - Cultured keratinocyte allografts from unrelated donors can be readily grown as sheets in large-scale cell culture and have been used as an immediate skin cover for severely burned patients. Despite the absence of passenger leukocytes and the unlimited amount of material that can be obtained for permanent skin coverage, the allografts are susceptible to rejection. Since MHC class I (MHCI) antigens serve as targets for allograft rejection, we investigated whether 'phenotypic knockout' of human MHCI could be achieved through expression of an ER-directed anti-human MHCI single-chain intrabody (sFvhMHCI) that is directed against a monomorphic, conformational epitope, expressed across species lines, on the MHCI heavy chain. Co-immunoprecipitation of both MHCI heavy chain and beta2 microglobulin occurred in transfected monkey COS-1 cells, while Jurkat T cells stably expressing the ER-directed sFvhMHCI intrabody showed that complete phenotypic knockout of MHCI cell surface expression could be achieved. Infection of several human cell lines of divergent tissue sources and different HLA haplotypes resulted in marked down-regulation of MHCI expression, even under conditions where inflammatory cytokines (eg gamma-IFN) which up-regulate MHCI expression were used. Finally, when adenovirus encoding the anti-human MHCI intrabody was used to transduce primary human keratinocytes, a marked reduction of surface MHCI expression was observed. These in vitro studies set the groundwork for in vivo studies to determine if intrabody-mediated knockout of MHCI can impair alloantigen expression and prolong the survival of keratinocyte allografts. PMID- 11938451 TI - Inhibition of retinal neovascularisation by gene transfer of soluble VEGF receptor sFlt-1. AB - Retinal angiogenesis is a central feature of the leading causes of blindness. Current treatments for these conditions are of limited efficacy and cause significant adverse effects. In this study, we evaluated the angiostatic effect of gene transfer of the soluble VEGF receptor sFlt-1 in a mouse model of ischaemia-induced retinal neovascularisation using adenovirus and adeno associated virus (AAV) vectors. We induced proliferative retinopathy in mice by exposure to 75% oxygen from postnatal day 7 (p7) to p12 and injected intravitreally recombinant viral vectors expressing the reporter green fluorescent protein (GFP) or vectors expressing the VEGF inhibitor sFlt-1. Efficient adenovirus-mediated GFP expression was evident in cells of the corneal endothelium and iris pigment epithelium. AAV-mediated GFP expression was evident in ganglion cells and cells of the inner nuclear layer of the retina. Vector mediated sFlt-1 expression was confirmed by ELISA of pooled homogenised whole eyes. Injection of either vector expressing sFlt-1 resulted in a reduction in the number of neovascular endothelial cells by 56% and 52% for adenovirus and AAV vectors, respectively (P < 0.05). Local gene transfer of sFlt-1 consistently inhibits experimental retinal neovascularisation by approximately 50% and offers a powerful novel approach to the clinical management of retinal neovascular disorders. PMID- 11938452 TI - LCR-mediated, long-term tissue-specific gene expression within replicating episomal plasmid and cosmid vectors. AB - Locus control regions (LCRs) are transcriptional regulatory elements, which possess a dominant chromatin remodelling and transcriptional activating capability conferring full physiological levels of expression on a gene linked in cis, when integrated into the host cell genome. Using the human beta-globin LCR (betaLCR) as a model, we show that this class of control element can drive high levels of tissue-specific gene expression in stably transfected cultured cells from within an Epstein-Barr virus-based plasmid REV. Furthermore, a 38-kb betaLCR minilocus-REV cosmid vector was efficiently retained and maintained therapeutic levels of beta-globin transgene expression in the absence of drug selective pressure over a 2-month period of continuous culture equivalent to at least 60 generations. This demonstrates for the first time the feasibility of using REVs for gene therapy of the haemoglobinopathies. Importantly, our results demonstrate that as in the case of integrated transgenes, expression from within REVs is prone to silencing but that the inclusion of the betaLCR prevented this repression of gene function. Therefore, appropriate control elements to provide and maintain tissue-specific gene expression, as well as the episomal status of REVs is a crucial feature in vector design. Our data suggest that LCRs can contribute to this vital function. PMID- 11938453 TI - Improved co-expression of multiple genes in vectors containing internal ribosome entry sites (IRESes) from human genes. AB - Incorporation of an internal ribosome entry site (IRES) into the gene therapy vector represents a promising strategy to efficiently co-express several gene products from the same promoter. However, vector systems that utilize the encephalomyocarditis virus IRES express the downstream gene much less efficiently than the upstream gene. In this study, we compared four IRESes isolated from human genes against the EMCV IRES, using beta-galactosidase and chloramphenicol acetyl transferase genes as reporters, to evaluate their potential for providing better expression of the downstream gene. We found that an IRES from the eukaryotic initiation factor 4G gene mediates greater than 100-fold higher expression of the downstream gene compared with the EMCV IRES in four different cell lines tested. Other mammalian IRESes displayed more variable results and smaller enhancement of downstream gene expression in three different cell lines tested. Furthermore, while the efficiency of the IRES from the vascular endothelium growth factor gene was not significantly higher than the EMCV IRES under normoxic conditions, expression was significantly increased under hypoglycemic conditions, suggesting that the VEGF IRES could be exploited in cancer gene therapy to preferentially target expression of therapeutic genes at the relatively hypoglycemic cores of tumors. PMID- 11938454 TI - An effective immunization and cancer treatment with activated dendritic cells transduced with full-length wild-type p53. AB - P53-based immunization is an attractive approach to cancer immunotherapy due to the accumulation of p53 protein in tumor, but not in normal cells. However, it was not known whether immune response against self-protein (p53) could be generated in vivo. Mouse dendritic cells (DCs) were transduced with adenoviral construct containing murine full-length wild-type p53 (Ad-p53). Repeated immunizations with these cells protected 60% of mice against challenge with MethA sarcoma cells bearing point mutations in p53 gene. Activation of DCs via ligation of CD40 significantly improved the results of immunization: all mice were protected against MethA sarcoma. The treatment of MethA tumor-bearing mice with activated Ad-p53-transduced DCs showed complete tumor rejection in four out of six mice. The specificity of antitumor immune response was confirmed by CTL assay. The analysis of phenotype and function of DCs demonstrated that the effect of CD40 ligation on these cells was enhanced by their infection with Ad-p53. The level of neutralizing anti-adenovirus antibody was moderately elevated in these mice. No signs of autoimmune reaction were evident during detailed pathological evaluation of treated mice. These data demonstrate that activated Ad-p53-infected DCs are able to break tolerance to this protein and can be used in immunotherapy of cancer. PMID- 11938455 TI - Growth inhibition of established B16-F10 lung metastases by sequential aerosol delivery of p53 gene and 9-nitrocamptothecin. AB - Growth inhibition of established tumor metastases in the lungs poses a difficult challenge for most clinical settings in spite of extensive multi-modality approaches. Aerosol delivery of drugs and genes holds promise for the treatment of disseminated lung metastases, since aerosol delivery can target the lungs specifically and uniformly. We previously demonstrated that aerosol delivery of dilauroylphosphatidylcholine liposome formulation of 9-nitrocamptothecin (9NC DLPC) inhibits B16-F10 melanoma lung metastases. Aerosol delivery of polyethleneimine-p53 DNA (PEI-p53) complexes results in a similar anti-tumor effect in the B16-F10 model. In both these previous studies, the protocols were designed to inhibit development of lung metastases. In this study we demonstrate, using the B16-F10 melanoma lung metastasis model, that sequential aerosol delivery of PEI-p53 and 9NC-DLPC acts additively to inhibit growth of established B16-F10 tumor metastases in the lungs. Mice injected with B16-F10 cells and treated with a combination of 9NC-DLPC (twice weekly) and PEI-p53 (once weekly) aerosol complexes starting on day 11 after tumor inoculation, exhibited a highly significant (P < 0.01) reduction in the number of visible tumor foci as compared with untreated mice or mice treated with either single agent alone, or with a combination of 9NC and a control plasmid. There was a highly significant reduction in the tumor burden, as well as the lung weights for the 9NC and p53 combination group (P < 0.001 as compared with other groups). Moreover, the doses of p53 gene and 9NC in the combination group were reduced at least two-fold as compared with our previous single agent studies, but still achieved significant tumor inhibition. Furthermore, the sequential aerosol delivery of p53 and 9NC lead to a 30-40% increase in the mean survival time of these mice, as compared with animals in different control groups. The data suggest that the combination of 9NC and p53 gene delivered by aerosol is an attractive strategy for growth inhibition of established tumor metastases in the lungs. PMID- 11938456 TI - Transient suppression of transgene expression by means of antisense oligonucleotides: a method for the production of toxin-transducing recombinant viruses. AB - Some of the therapeutic genes to be delivered by means of recombinant adenoviruses code for toxic compounds. Expression of these sequences can be deleterious to the complementation cells used for vector production, making it often difficult to generate high-titer stocks of toxin-transducing recombinant adenoviruses. In this work, we present a novel strategy for the transient post transcriptional down-regulation of toxic transgene expression during the vector production phase, through the administration of phosphorothioate-modified antisense oligodeoxyribonucleotides. This method was successfully applied to the production of hybrid adenoviruses that contain the gene encoding the cytotoxic parvoviral protein NS1. The generation of recombinant adenoviruses in 293T cells was found to be fully suppressed as a result of adding of the NS gene to the vector genome. Yet, the production of NS-harboring hybrid adenoviruses could be rescued by treating the producer cells with antisense oligonucleotides specific for the translation initiation region of the NS transcript. This rescue correlated with a striking reduction of NS RNA and protein levels in the complementation cells. These data provide proof of principle of the suitability of the antisense oligonucleotides strategy for overcoming the interference of harmful transgenes with the production of adenoviral and other vectors. PMID- 11938457 TI - Inhibition of HIV-1 replication by novel lentiviral vectors expressing transdominant Rev and HIV-1 env antisense. AB - Retroviral vectors expressing transdominant negative mutants of Rev (TdRev) inhibit HIV-1 replication by preventing the nuclear export of unspliced viral transcripts, thus inhibiting the synthesis of Gag-Pol, Env and reducing the levels of genomic RNA available for packaging. Due to these effective mechanisms of inhibition, production of HIV-1-based lentiviral vectors expressing TdRev has been difficult. Here we describe HIV-based vectors in which expression of TdRev is negatively regulated by Rev expression. In these vectors, we maintained the wild-type HIV-1 Tat/Rev exons and intron configuration and its mode of splicing regulation. The second Rev exon was mutated to encode TdRev. Inhibition of TdRev expression by Rev during vector production yields high titer vector preparations. A second vector containing an additional anti-HIV gene (env-antisense) was constructed by flipping a 1.2-kb env fragment contained within the Tat/TdRev intron. SupT1 cells and primary CD4+ lymphocytes transduced with these vectors inhibit HIV-1 replication and show a preferential advantage for survival. Although these vectors are poorly mobilized to secondary target cells by wild type HIV-1, they reduce the infectivity of the wild-type virions escaping inhibition. PMID- 11938458 TI - Syngeneic central nervous system transplantation of genetically transduced mature, adult astrocytes. AB - Advances in the development of highly infectious, replication-deficient recombinant retroviruses provide an efficient means of stable transfer of gene expression. Coupled with ex vivo transduction, surrogate cell populations can be readily implanted into the brain, thus serving as vehicles for delivering selected gene products into the central nervous system (CNS). Here we report that rat astrocytes can be routinely and safely isolated from brain tissue of a living donor by use of short-term gelatin sponge implants. The mature, nontransformed astrocytes were easily expanded, maintained in long-term tissue cultures and were efficiently transduced with an amphotropic retrovirus harboring a heterologous, fused transgene. In vitro retroviral infection rendered the nontransformed cells essentially 100% viable after exposure. The level of efficiency of infection (30 50% effective genome integration of provirus and expression of transgene in target cell populations) and minimal cell toxicity obviated the need to harvest large numbers of target cells. Cultured transduced astrocytes were resilient and exhibited select peptide expression for up to 1 year. Subsequently, transduced astrocytes were used in a series of experiments in which cells were transplanted intracerebrally in syngeneic animals. Post-implantation, astrocytes seeded locally and either insinuated into the surrounding parenchyma in situ or exhibited a variable degree of migration, depending on the anatomic source of astrocytes and the targeted brain implantation site. Transduced astrocytes remained viable in excess of 8 months post-transplantation and exhibited sustained transgenic peptide expression of green fluorescent protein/neomycin phosphotransferase in vivo. The sequential isolation and culture of nontransformed, mature, adult astrocytes and recombinant retrovirus-mediated transduction in vitro followed by brain reimplantation represents a safe and effective means for transferring genetic expression to the CNS. This study lays the foundation for exploring the utility of using a human autologous transplantation system as a potential gene delivery approach to treat neurological disorders. Prepared and utilized in this manner, autologous astrocytes may serve as a vehicle to deliver gene therapy to the CNS. PMID- 11938459 TI - Ablation of microvessels in vivo upon dimerization of iCaspase-9. AB - Anti-angiogenic therapies based on targeted disruption of the tumor microvascular network have been proposed for cancer treatment. Inhibitors of the endothelial cell pro-survival pathway mediated by VEGF were shown to activate caspases and cause microvascular regression, but the efficacy of this strategy can be hindered by the engagement of redundant survival pathways. Alternatively, if direct activation of an apical pro-apoptotic caspase is sufficient to disrupt microvessels in vivo, such a strategy could potentially override upstream endothelial cell survival inputs and disrupt tumor neovascular networks. Here, we fused caspase-9 to a mutated FKBP12 domain to express an inducible caspase-9 molecule (iCaspase-9) that can be activated by a cell-permeable dimerizer drug, and transduced this construct into primary endothelial cells. We found that drug induced dimerization of iCaspase-9 is sufficient to activate endogenous caspase-3 and trigger apoptosis even when endothelial cells are treated with the pro survival factors VEGF or bFGF. A single intraperitoneal injection of the dimerizer drug induced apoptosis of endothelial cells expressing iCaspase-9 and elimination of human microvessels engineered in immunodeficient mice. These results demonstrate that the activation of iCaspase-9 disrupts microvessels in vivo, and suggest a novel anti-angiogenic strategy based on the expression and controlled activation of an inducible death gene in neovascular endothelial cells. PMID- 11938460 TI - Human cytidine deaminase as an ex vivo drug selectable marker in gene-modified primary bone marrow stromal cells. AB - Naturally occurring drug resistance genes of human origin can be exploited for selection of genetically engineered cells co-expressing a desired therapeutic transgene. Their non-immunogenicity in clinical applications would be a major asset. Human cytidine deaminase (hCD) is a chemoresistance gene that inactivates cytotoxic cytosine nucleoside analogs, such as cytosine arabinoside (Ara-C). The aim of this study was to establish if the hCD gene can serve as an ex vivo dominant selectable marker in engineered bone marrow stromal cells (MSCs). A bicistronic retrovector comprising the hCD cDNA and the green fluorescent protein (GFP) reporter gene was generated and used for transduction of A549 cells and primary murine MSCs. Analysis of transduced cells demonstrated stable integration of proviral DNA, more than 1000-fold increase in CD enzyme activity, and drug resistance to cytosine nucleoside analogs. In a mixture of transduced and untransduced MSCs, the percentage of retrovector-expressing cells could be increased to virtual purity (>99.5%) through in vitro drug selection with 1 microM Ara-C. Increased selective pressure with 2.5 microM Ara-C allowed for enrichment of a mixed population of MSCs expressing approximately six-fold higher levels of GFP and of CD activity when compared with unmanipulated engineered MSCs. Moreover, engraftment and endothelial differentiation of these in vitro selected and enriched gene-modified marrow stromal cells was demonstrated by Matrigel assay in vivo. In conclusion, these findings outline the potential of human CD as an ex vivo selection and enrichment marker of genetically engineered MSCs for transgenic cell therapy applications. PMID- 11938461 TI - Gene delivery to the lung using protein/polyethylenimine/plasmid complexes. AB - Delivery of genes to the lung has enormous potential in a wide variety of illnesses, from lung cancer to genetic deficiency diseases. Many delivery systems have been utilized, each with its own advantages and limitations. Polyethylenimine is a polycation capable of binding and compacting DNA, enabling intravascular plasmid delivery to normal tissues in such a way that the plasmid can be expressed in a proportion of the exposed cells. We have developed a novel intravenous method to deliver small amounts of plasmid to lung tissue, using nontoxic quantities of polyethylenimine in combination with albumin (or other soluble proteins). Injection of 1 microg or less of plasmid resulted in highly efficient gene expression in lung interstitial and endothelial tissues (0.5 to 1 ng luciferase per microg plasmid DNA), while larger quantities of plasmid reduced relative gene expression. Using luciferase as a reporter gene, single injections had maximal gene expression between 24 and 48 h, with a rapid decline thereafter. In contrast to some other delivery systems, however, no inhibition of gene expression occurred during multiple rounds of plasmid administration through 20 days. As a result, this method may have useful applications in diseases that could benefit from recurrent therapeutic gene delivery. PMID- 11938462 TI - Replication and packaging of helper-dependent adenoviral vectors. AB - A helper-dependent adenovirus vector (HD AdV) that lacks all the coding sequences of viral proteins from the vector backbone was developed to overcome the problem of host cellular immune responses against E1-deleted AdV. One of the limitations of the current HD AdV system is its relatively inefficient propagation compared with that of E1-deleted AdV, which deters application of the HD AdV, especially in large animal models. We hypothesized that the low titers might be due in part to defects in replication and packaging of the vector DNA. We propagated two HD AdVs with similar marker gene cassettes and stuffer sequences, using two different helper viruses, and determined the replication and packaging efficiencies of viral DNA. Our analysis indicated a difference in replication and packaging efficiencies between the two vectors, which resulted in different propagation efficiencies. Furthermore, dl309, which is similar to the wild-type virus, demonstrated superior helper function over that of the loxP-containing helper virus, AdLC8cluc. These findings may have significant implications for the design of improved production systems of HD AdVs. PMID- 11938463 TI - Lactic acid bacteria and yeasts in kefir grains and kefir made from them. AB - In an investigation of the changes in the microflora along the pathway: kefir grains (A)-->kefir made from kefir grains (B)-->kefir made from kefir as inoculum (C), the following species of lactic acid bacteria (83-90%) of the microbial count in the grains) were identified: Lactococcus lactis subsp. lactis, Streptococcus thermophilus, Lactobacillus delbrueckii subsp. bulgaricus, Lactobacillus helveticus, Lactobacillus casei subsp. pseudoplantarum and Lactobacillus brevis. Yeasts (10-17%) identified were Kluyveromyces marxianus var. lactis, Saccharomyces cerevisiae, Candida inconspicua and Candida maris. In the microbial population of kefir grains and kefir made from them the homofermentative lactic streptococci (52-65% and 79-86%, respectively) predominated. Within the group of lactobacilli, the homofermentative thermophilic species L. delbrueckii subsp. bulgaricus and L. helveticus (70-87% of the isolated bacilli) predominated. Along the pathway A-->B-->C, the streptococcal proportion in the total kefir microflora increased by 26-30% whereas the lactobacilli decreased by 13-23%. K. marxianus var. lactis was permanently present in kefir grains and kefirs, whereas the dominant lactose-negative yeast in the total yeast flora of the kefir grains dramatically decreased in kefir C. PMID- 11938464 TI - Ensiling whole-crop wheat and corn in large containers with Lactobacillus plantarum and Lactobacillus buchneri. AB - The effect of applying Lactobacillus buchneri, alone or in combination with Lactobacillus plantarum, at ensiling, on the aerobic stability of wheat and corn silages was studied in 50-l plastic containers. Treatments comprised control (no additives), L. plantarum, L. buchneri and a combination of L. plantarum+L. buchneri. After 3 months of storage, the wheat silages treated with L. buchneri had higher acetic acid contents than the control or L. plantarum-treated silages, and were free of mold, whereas the top layers of the control or L. plantarum treated silages were moldy. In an aerobic stability test the L. buchneri-treated silages were stable, whereas those treated with L. plantarum deteriorated. In the corn silages the effects of L. buchneri were not as clear and the top layer was moldy in all silages. However, L. buchneri also improved the aerobic stability of the corn silage, as indicated by lower yeast numbers, less CO(2) production and stable pH. It is concluded that L. buchneri has a potential as a silage additive that protects the silage upon aerobic exposure. The 50-l plastic containers can serve as an appropriate model to test silage additives before conducting full scale farm experiments. PMID- 11938465 TI - Production of a novel FK520 analog in Streptomyces hygroscopicus: improving titer while minimizing impurities. AB - FK520, also called ascomycin, is an immunosuppressive agent produced by Streptomyces hygroscopicus. Engineering the polyketide synthase genes of the parent strain generated novel FK520 analogs with the potential for improved in vivo stability. By replacing the acyl transferase (AT) domain in the polyketide synthase module 8 with an AT specific for methylmalonyl CoA (the rapamycin AT 3), the strain produced 13-desmethoxy-13-methyl-FK520 (13dmmFK520). Process development and scale-up studies of this recombinant S. hygroscopicus strain producing 13dmmFK520 are described here. Production kinetics and compound stability in fermentation broth were significantly different compared to the native FK520. Fermentation of the new strain resulted in the synthesis of a contaminating substance that co-purified with the 13dmmFK520. To optimize 13dmmFK520 production and to facilitate purification, growth parameters and media development were examined. Although a medium was identified that increased product titers by ca. 300%, the ratio of impurity to product was doubled. Lower dissolved oxygen (20% compared to 50% and 80%) increased titers by 20% with no appreciable effect on the concentration of impurity. Increasing the fermentation pH from 6.0 to 6.5 did not change the 13dmmFK520 titer, but reduced the impurity to-product ratio by approximately 450%. PMID- 11938467 TI - Halophiles. PMID- 11938466 TI - Modulation of epothilone analog production through media design. AB - Recently, the epothilone polyketide synthase (PKS) was successfully introduced into a heterologous production host for the large-scale production of epothilone D. We have found that at least three other epothilones can also be produced as the major fermentation product of this recombinant strain by supplementation of specific substrates to the production media. Addition of acetate or propionate to the media results in modulation of the epothilone D:C ratio, whereas addition of L-serine with either acetate or propionate yields epothilone H(1) or H(2) as the major product. This strategy permits production of at least four novel epothilones by culturing a single host with a genetically modified epothilone PKS in various media. PMID- 11938468 TI - Halocins and sulfolobicins: the emerging story of archaeal protein and peptide antibiotics. AB - Production of antibiotic peptides and proteins is a near-universal feature of living organisms regardless of phylogenetic classification. Bacteriocins (proteinaceous antimicrobials from the domain Bacteria) have been studied for over 75 years, and the eucaryocins (proteinaceous antimicrobials from the domain Eucarya) since the early 1960s. However, one domain of organisms, the Archaea, containing hyperthermophiles, extreme halophiles and the methanogens, is just beginning to be scrutinized for the production of peptide antibiotics. Production of archaeal proteinaceous antimicrobials (archaeocins) from extreme halophiles (halocins) is a nearly universal feature of the rod-shaped haloarchaea. Halocin activity is first detectable in culture supernatants at the beginning of the transition into stationary phase, concomitant with an induction of transcription of the structural gene. Halocins are diverse in size, consisting of proteins as large as 35 kDa and peptide "microhalocins" as small as 3.6 kDa. The 36 amino acids of microhalocin HalS8 are located in the interior of a 311-residue pro protein from which they are liberated by an unknown mechanism. Microhalocins are hydrophobic and robust, withstanding heat, desalting and exposure to organic solvents. Unlike the peptide bacteriocins and the eucaryocins, microhalocins possess a large number of neutral residues and are not cationic, leaving their mechanism(s) of action mostly a mystery. While microhalocins affect a variety of haloarchaeal genera (kingdom Euryarchaeota), they also exhibit cross-kingdom toxicity, inhibiting or killing Sulfolobus species (kingdom Crenarchaeota). Finally, archaeocins also are produced by the hyperthermophile "Sulfolobus islandicus". These 20-kDa protein antibiotics are not excreted into the environment, but are associated with small particles apparently derived from the cell's S-layer. PMID- 11938469 TI - The question of uniqueness of ancient bacteria. AB - Microorganisms are associated with a variety of ancient geological materials. However, conclusive proof that these organisms are as old as the geological material and not more recent introductions has generally been lacking. Over the years, numerous reports of the isolation of ancient bacteria from geological materials have appeared. Most of these have suffered from the fact that the protocol for the surface sterilization of the sample was either poorly defined, inadequate or rarely included data to validate the overall effectiveness of the sterilization protocol. With proper sterility validation and isolation protocol, a legitimate claim for the isolation of an ancient microbe can be made. Biochemical, physiological, or morphological data indicate that these ancient microbes are not significantly different from modern isolates. As the role (decomposition) of modern and ancient microbes has not changed over time, it is probably unreasonable to expect these organisms to be vastly different. A discussion on the reasons for the homogeneity of ancient and modern microbes is presented. PMID- 11938470 TI - Industrial microbiology of solar salt production. AB - Solar salterns can be modeled as giant outdoor chemostats, much like a series of dams on a slow-moving river. Microorganisms and their products play an essential, but sometimes uncharacterized, role in salt production in these ponds, from seawater salinity up through NaCl saturation. They may physically affect the evaporation process and their by-products may chemically modify or bind with dissolved ions. Many solar salt facilities engage microbiologists to establish monitoring programs for analyses of nutrients, standing crop and associated biological variables in the ponds. Other solar salt companies engage microbiologists only when there are "crises" in the ponds that interfere with salt production. PMID- 11938472 TI - Diversity of halophilic microorganisms: environments, phylogeny, physiology, and applications. AB - The phylogenetic diversity of microorganisms living at high salt concentrations is surprising. Halophiles are found in each of the three domains: Archaea, Bacteria, and Eucarya. The metabolic diversity of halophiles is great as well: they include oxygenic and anoxygenic phototrophs, aerobic heterotrophs, fermenters, denitrifiers, sulfate reducers, and methanogens. The diversity of metabolic types encountered decreases with salinity. The upper salinity limit at which each dissimilatory process takes place is correlated with the amount of energy generated and the energetic cost of osmotic adaptation. Our understanding of the biodiversity in salt-saturated environments has increased greatly in recent years. Using a combination of culture techniques, molecular biological methods, and chemotaxonomic studies, we have obtained information on the nature of the halophilic Archaea as well as the halophilic Bacteria that inhabit saltern crystallizer ponds. Several halophilic microorganisms are being exploited in biotechnology. In some cases, such as the production of ectoine, the product is directly related to the halophilic behavior of the producing microorganism. In other cases, such as the extraction of beta-carotene from Dunaliella or the potential use of Haloferax species for the production of poly-beta hydroxyalkanoate or extracellular polysaccharides, similar products can be obtained from non-halophiles, but halophilic microorganisms may present advantages over the use of non-halophilic counterparts. PMID- 11938471 TI - Microbial diversity and complexity in hypersaline environments: a preliminary assessment. AB - The microbial communities in solar salterns and a soda lake have been characterized using two techniques: BIOLOG, to estimate the metabolic potential, and amplicon length heterogeneity analysis, to estimate the molecular diversity of these communities. Both techniques demonstrated that the halophilic Bacteria and halophilic Archaea populations in the Eilat, Israel saltern are dynamic communities with extensive metabolic potentials and changing community structures. Halophilic Bacteria were detected in Mono Lake and the lower salinity ponds at the Shark Bay saltern in Western Australia, except when the crystallizer samples were stressed by exposure to Acid Green Dye #9899. At Shark Bay, halophilic Archaea were found only in the crystallizer samples. These data confirm both the metabolic diversity and the phylogenetic complexity of the microbial communities and assert the need to develop more versatile media for the cultivation of the diversity of bacteria in hypersaline environments. PMID- 11938473 TI - Workbook in Practical Neonatology. PMID- 11938476 TI - Cancer Research Foundation of America. PMID- 11938475 TI - Juliana Denekamp: An Appreciation. PMID- 11938478 TI - Children born after assisted reproductive technology. AB - Since the birth of Louise Brown in July 1978 and the birth of the first intracytoplasmic sperm injection (ICSI) child in January 1992 many couples with longstanding female-factor or male-factor infertility can be helped to overcome their infertility resulting in a delivery and birth of a child. The final and ultimate goal of all infertility treatments has been to give the large population of infertile couples a chance to fulfil their childwish and experience the happiness of having a healthy child. Major advances have been made in the different treatment protocols for infertility during the last 25 years. It is, however, surprising that only a limited number of studies have been carried out assessing the health of the children born after ART. In this review we shall comment on the limitations of follow-up studies on ART children and we shall review existing data on the outcome of in vitro fertilization (IVF) and ICSI pregnancies. The most important outcome data consist of information on minor and major congenital malformations obtained prenatally or after birth, as well as on the further development of the children. PMID- 11938479 TI - Unlicensed and off-label medication use in a neonatal intensive care unit: a prospective study. AB - The aim of this study is to determine the extent of use of medications that have not been specifically licensed for use in children (unlicensed), or medications whose use is not in accordance with the conditions of their license (off-label), in neonates in a neonatal intensive care unit (NICU). Medications given to 105 neonates were prospectively reviewed every 2 weeks during a 4-month period. The assessment as to whether every medication prescribed was unlicensed or off-label for use in children was based on a number of reference sources. Five hundred and twenty-five series of medications were used, of which 310 (59%) were off-label and 87 (16%) were unlicensed. Ninety-eight neonates (93%) received at least one off-label medication. The major reason for prescribing off-label medications was a deviation from the recommended dosage or age of the patient. The reason for giving unlicensed medications was changes in the formulation of the medication. The use of off-label and unlicensed medications is common in neonates. In view of the gravity and problematic nature of the issue, international consensus is evolving to conduct clinical trials in neonates and infants, with regard to medications already on the market, and new medications. PMID- 11938480 TI - Fetal echocardiographic screening of pregnancies of mothers with anti-Ro and/or anti-La antibodies. AB - The objectives of this study are to assess the incidence of congenital complete heart block (CCHB) in pregnant women who are anti-Ro and/or La positive. Between January, 1988 and July 1997, 118 pregnancies in 105 women were assessed by fetal echo at 18, 24, and 32 weeks' gestation. Of these 105, 96 had no history of a previous fetus with CCHB; 11(12 pregnancies) a history of a pregnancy with CCHB; and 4 a previous child with cutaneous neonatal lupus erythematosus (CNLE). The 102 pregnancies in 96 women with anti-Ro and/or anti-La antibodies and no history of a previous child with NLB, resulted in 100 live-births with no CCHB. There was 1 dilated cardiomyopathy at 6 months, 1 child with CCHB, and 1 with sclerosis of the endocardium. No case of CCHB was observed in 102 pregnancies without a previous history of CCHB. These results suggest that the risks of CCHB, without a prior history are low. PMID- 11938481 TI - Contribution of elective delivery to severe respiratory distress at term. AB - We sought to determine the contribution of elective delivery to severe respiratory distress syndrome (RDS) on a weekly basis from 37-40 weeks' gestation. Chart reviews confirmed gestational age, delivery reason, and primary diagnosis of all inborn neonates with RDS requiring mechanical ventilation delivered at 37 0/7-40 6/7 weeks' gestation from 1/1/90-12/31/99. Exclusion criteria were sepsis, pneumonia, meconium aspiration, asphyxia, pulmonary hemorrhage, hydrops, chromosomal abnormality, or congenital malformations affecting respiration. Thirty-five thousand and thirty-one deliveries occurred from 37 0/7-40 6/7 weeks; 18 (0.05%) had RDS requiring mechanical ventilation. Nine infants delivered at 37 0/7-37 6/7 weeks, (OR for RDS = 38.5; 95% CI = 8.3, 178.3), seven delivered at 38 0/7-38 6/7 weeks, (OR for RDS = 13.3; 95% CI = 2.8, 64.0), and two delivered at 39 0/7-40 6/7 weeks. Six of 18 infants were electively delivered without documented lung maturity. Infants born at 37 0/7-38 6/7 weeks are at significantly increased risk for severe RDS. One third of RDS cases were potentially avoidable. PMID- 11938482 TI - Detection of Helicobacter pylori seropositivity in hyperemesis gravidarum and correlation with symptoms. AB - The aim of this study was to investigate whether there was a relationship between H. pylori (Helicobacter pylori) infection positivity and characteristics (time and duration) of hyperemesis gravidarum (HG) symptoms. Forty-seven pregnant women with HG and 39 pregnant controls at the same gestational weeks without any gastrointestinal symptoms were included in this prospective study. H. pylori serum Immunoglobulin (Ig) G concentrations were determined by enzyme-linked immunoadsorbent assay (ELISA) in patients with HG and controls. Positive serology for H. pylori was correlated with the duration of symptoms in patients with HG. The prevalence of H. pylori infection was 85.1% (40 of 47) and 64.1% (25 of 39) in patients with HG and controls, respectively (p <0.05, chi (2) test). Mean values of H. pylori IgG (+/- standard deviation) were significantly higher in patients with HG than in controls (22.66 aIU/mL +/- 22.34 vs. 11.54 aIU/mL +/- 13.89, p <0.01, Student's t-test). In HG group, time (gestational weeks) and the duration (weeks) of HG symptoms for patients serologically positive and negative for H. pylori were 6.95 +/- 1.55 versus 6.58 +/- 1.78 weeks, (p >0.05, Student's t-test) and 8.35 +/- 5.28 versus 11.40 +/- 7.17 weeks (p >0.05, Mann-Whitney U test), respectively. There was no correlation between duration of HG symptoms and serum H. pylori IgG concentrations. Although a majority of pregnant women with HG were serologically positive for H. pylori infection, there was no correlation between positive serology and duration of symptoms, which is not suggestive of a direct causal relationship between H. pylori infection and HG. PMID- 11938483 TI - Hyperemesis gravidarum is associated with oxidative stress. AB - Hyperemesis gravidarum (HEG), associated with pregnancy, is a severe form of nausea and vomiting causing decrease in nutrient antioxidants. Hence, we hypothesize that oxidation injury may be involved in the pathogenesis of HEG. Plasma levels of the ubiquitous antioxidant, reduced glutathione (GSH) may serve as a sensitive measure for systemic oxidative stress. Women with pregnancies complicated by HEG (study group) were compared with pregnant women without HEG (pregnant control group) and with healthy nonpregnant women (nonpregnant control group). Plasma GSH levels were determined in the study group at the time of admission to hospital, and when the vomiting had ceased, it was compared with those of the two control groups. Plasma GSH levels were significantly higher in the pregnant control group than in nonpregnant controls (6.13 +/- 2.9 microM vs. 1.01 +/- 0.3 microM p <0.01). In contrast, values in the HEG women at the time of admission were significantly lower than the pregnant controls (3.12 +/- 1.6 microM, p <0.01). At the second sampling, when the women had ceased vomiting, plasma GSH values were higher than at the acute stage of the illness and were no longer significantly different from the pregnant control group (4.43 +/- 1.6 microM). Low values of plasma GSH in HEG patients suggest that oxidative stress is associated with this condition. PMID- 11938484 TI - Down syndrome: still a social stigma. AB - Down Syndrome (DS) is a commonly occurring chromosomal abnormality. The incidence increases with advancing maternal age over 35 years. Over the last three decades, tremendous progress has been made in the medical and surgical treatment of these infants. Nationally, a great deal of resources are allocated to DS infants to improve their growth and development. Yet, the perception remains that the DS infant is still not openly accepted by parents and society, as illustrated by the presented cases. This lack of acceptance creates many complex ethical challenges in treating such babies, starting with fetal diagnosis of the disorder in the womb and moving through early stages after the birth of the baby. We argue that health professionals have the responsibility to help make public attitudes more accepting of Down Syndrome. Professionals should encourage social and community involvement of these children. The National Association for Down Syndrome should be contacted periodically to promote activities to enhance public awareness. To the end of greater acceptance, we suggest a ritual at birth that might improve the acceptance of the DS child into the family and the community and hence help improve social attitudes toward Down Syndrome. PMID- 11938485 TI - Reference values for lactic dehydrogenase activity and isoenzyme distribution in cerebrospinal fluid in neonates with fever but no evidence of cerebral disease. AB - Levels of lactic dehydrogenase (LDH) in the cerebrospinal fluid (CSF) have been used to evaluate a variety of neurologic disorders. However, few studies have been performed on neonates. CSF samples were collected from 39 neonates, 19 aged 1-4 weeks and 20 aged 5-8 weeks with neonatal fever. Findings were compared to those in 15 infants mean age 10 +/- 6.4 months with suspected nervous system disease and normal CSF findings. Mean total LDH activity in the CSF was significantly higher in the first month of life than in the second month of life and compared with the control group (p <0.001). The LDH isoenzyme pattern was also different, with lower values of LDH-1 than LDH-2 and -3 in the study groups. Differences were significant among all three groups for LDH-1 (p <0.001) and between the whole study group and the controls for LDH-2 and -3 (p <0.001). We propose reference values for total LDH activity and LDH isoenzyme distribution in the CSF of normal neonates. PMID- 11938487 TI - Pattern-based approaches to phonological therapy. AB - Pattern-based approaches to phonological disorders emerged in the 1970s and 1980s after researchers realized that phonological "rules" used to describe speech patterns of adults could be used in the analysis of children's speech. In this article, a pattern-based approach to assessment and treatment of phonological disorders is described, based on identification and treatment of phonological processes. Means of identifying and quantifying phonological processes are provided and assessment tests are described. Principles underlying the use of phonological processes in intervention programs are presented and decisions regarding target selection are discussed. A case study of the treatment program for one child illustrates the application of phonological processes within a cycles approach. PMID- 11938488 TI - Are oral-motor exercises useful in the treatment of phonological/articulatory disorders? AB - The utility of oral-motor exercises in the remediation of children's speech acquisition delays continues to be a controversial issue. There are few empirical evaluations of the efficacy of these nonspeech activities in effecting speech changes, although much can be learned from investigations in related fields. The purpose of this article is to review the extant studies of the relation between oral-motor exercises and speech production in children as well as to examine the motor learning literature to gain a broader perspective on the issue. Results of this examination lead to questions about the procedures that are currently applied as well as to suggestions for future development of nonspeech activities in the treatment of children's phonological/articulatory disorders. PMID- 11938489 TI - From phonological therapy to phonological awareness. AB - Children with speech difficulties often have delayed phonological awareness development and associated literacy problems. Speech-language pathologists (S LPs) typically use phonological and articulatory approaches in their treatment of such children. However, it is unclear to what extent phonological awareness training, originally designed to promote literacy skills, might also improve children's speech output. This article adopts a psycholinguistic approach to examine the nature and development of phonological awareness and to explore the relationship between phonological awareness training and phonological therapy. The role of phonological awareness in predicting literacy development in children is discussed, and principles for analyzing the psycholinguistic properties of therapy tasks are presented. Phonological awareness cannot be dealt with independently as it is an integral part of articulation and phonological intervention. Further, phonological awareness is a necessary "on-line" skill in the dynamic communication process between therapist and child. Failure to take this into account will result in inappropriately targeted therapy and pragmatic breakdown between the child and S-LP. PMID- 11938490 TI - Phonotactic therapy. AB - Words derive their structure not only from the sounds they include but also from the organization of those sounds within the word. This organization is the phonotactic level of the word: roughly, its shape including the sequence of its elements. Often, children with immature or disordered phonologies demonstrate phonotactic as well as phonetic limitations. Sometimes, the child may produce an age-appropriate variety of consonants and vowels but be unable to use them in the configurations required by the language: final consonants, clusters, multisyllabic words, and so forth. In such cases, the most appropriate therapy goals may be phonotactic, rather than phonetic, ones. Studies have shown that clinical focus on a new word or syllable shape may generalize well beyond the specific sound or sounds targeted in that position. These ideas are explored in this article, along with specific therapy results and recommendations for various phonotactic limitations. PMID- 11938491 TI - Minimal pair approaches to phonological remediation. AB - This article considers linguistic approaches to phonological remediation that emphasize the role of the phoneme in language. We discuss the structure and function of the phoneme by outlining procedures for determining contrastive properties of sound systems through evaluation of minimal word pairs. We then illustrate how these may be applied to a case study of a child with phonological delay. The relative effectiveness of treatment approaches that facilitate phonemic acquisition by contrasting pairs of sounds in minimal pairs is described. A hierarchy of minimal pair treatment efficacy emerges, as based on the number of new sounds, the number of featural differences, and the type of featural differences being introduced. These variables are further applied to the case study, yielding a range of possible treatment recommendations that are predicted to vary in their effectiveness. PMID- 11938492 TI - Language-based intervention for phonological disorders. AB - Children with phonological disorders often display difficulty in other domains of language. Language-based approaches focus on all aspects of language; therefore, little attention may be drawn to sound errors and these may not be specific targets of intervention. These approaches involve a variety of naturalistic, conversationally based techniques such as focused stimulation in the form of expansions and recasts, scaffolding narratives, and elicited production devices such as forced choice questions, cloze tasks, and preparatory sets. Results from well-controlled group studies are inconclusive regarding the cross-domain effects of morphosyntax approaches on phonology. There are, however, individual children whose phonology improves with a language-based approach. Preliminary evidence suggests that such an approach may be an appropriate choice for children with both speech and language impairments whose phonological systems are highly inconsistent. One advantage of a language-based approach is that it may lead to simultaneous improvements in both speech and language for children with difficulty in both these domains. It is also a viable option when service delivery dictates classroom and collaborative settings. When a language-based approach is chosen for children with phonological disorders, it is imperative that the practitioner monitor phonological progress closely to ensure its effectiveness. PMID- 11938494 TI - The TRIM37 gene encodes a peroxisomal RING-B-box-coiled-coil protein: classification of mulibrey nanism as a new peroxisomal disorder. AB - Mulibrey nanism is a rare growth disorder of prenatal onset caused by mutations in the TRIM37 gene, which encodes a RING-B-box-coiled-coil protein. The pathogenetic mechanisms of mulibrey nanism are unknown. We have used transiently transfected cells and antibodies raised against the predicted TRIM37 protein to characterize the TRIM37 gene product and to determine its intracellular localization. We show that the human TRIM37 cDNA encodes a peroxisomal protein with an apparent molecular weight of 130 kD. Peroxisomal localization is compromised in mutant protein representing the major Finnish TRIM37 mutation but is retained in the protein representing the minor Finnish mutation. Colocalization of endogenous TRIM37 with peroxisomal markers was observed by double immunofluorescence staining in HepG2 and human intestinal smooth muscle cell lines. In human tissue sections, TRIM37 shows a granular cytoplasmic pattern. Endogenous TRIM37 is not imported into peroxisomes in peroxin 1 (PEX1(-/ )) and peroxin 5 (PEX5(-/-)) mutant fibroblasts but is imported normally in peroxin 7 (PEX7(-/-)) deficient fibroblasts, giving further evidence for a peroxisomal localization of TRIM37. Fibroblasts derived from patients with mulibrey nanism lack C-terminal TRIM37 immunoreactivity but stain normally for both peroxisomal matrix and membrane markers, suggesting apparently normal peroxisome biogenesis in patient fibroblasts. Taken together, this molecular evidence unequivocally indicates that TRIM37 is located in the peroxisomes, and Mulibrey nanism thus can be classified as a new peroxisomal disorder. PMID- 11938496 TI - Nipah virus encephalitis outbreak in Malaysia. AB - Emerging infectious diseases involving zoonosis have become important global health problems. The 1998 outbreak of severe febrile encephalitis among pig farmers in Malaysia caused by a newly emergent paramyxovirus, Nipah virus, is a good example. This disease has the potential to spread to other countries through infected animals and can cause considerable economic loss. The clinical presentation includes segmental myoclonus, areflexia, hypertension, and tachycardia, and histologic evidence includes endothelial damage and vasculitis of the brain and other major organs. Magnetic resonance imaging has demonstrated the presence of discrete high-signal-intensity lesions disseminated throughout the brain. Nipah virus causes syncytial formation in Vero cells and is antigenically related to Hendra virus. The Island flying fox (Pteropus hypomelanus; the fruit bat) is a likely reservoir of this virus. The outbreak in Malaysia was controlled through the culling of >1 million pigs. PMID- 11938495 TI - Reduced-median-network analysis of complete mitochondrial DNA coding-region sequences for the major African, Asian, and European haplogroups. AB - The evolution of the human mitochondrial genome is characterized by the emergence of ethnically distinct lineages or haplogroups. Nine European, seven Asian (including Native American), and three African mitochondrial DNA (mtDNA) haplogroups have been identified previously on the basis of the presence or absence of a relatively small number of restriction-enzyme recognition sites or on the basis of nucleotide sequences of the D-loop region. We have used reduced median-network approaches to analyze 560 complete European, Asian, and African mtDNA coding-region sequences from unrelated individuals to develop a more complete understanding of sequence diversity both within and between haplogroups. A total of 497 haplogroup-associated polymorphisms were identified, 323 (65%) of which were associated with one haplogroup and 174 (35%) of which were associated with two or more haplogroups. Approximately one-half of these polymorphisms are reported for the first time here. Our results confirm and substantially extend the phylogenetic relationships among mitochondrial genomes described elsewhere from the major human ethnic groups. Another important result is that there were numerous instances both of parallel mutations at the same site and of reversion (i.e., homoplasy). It is likely that homoplasy in the coding region will confound evolutionary analysis of small sequence sets. By a linkage-disequilibrium approach, additional evidence for the absence of human mtDNA recombination is presented here. PMID- 11938497 TI - The 1998 enterovirus 71 outbreak in Taiwan: pathogenesis and management. AB - The most recently discovered enterovirus, enterovirus 71 (EV71), is neurotropic and may cause severe disease and sudden death in children. In 1998, a large outbreak of enterovirus infection occurred in Taiwan that resulted in 405 severe cases in children and 78 deaths. Of the 78 children who died, 71 (91%) were <5 years old. EV71 was the primary agent in fatal cases of infection. Most of these patients died within 1-2 days of admission to the hospital. We hypothesize that EV71 directly attacks the central nervous system and causes neurogenic pulmonary edema and cardiac decompensation through the mechanism of sympathetic hyperactivity and inflammatory responses. Early recognition of risk factors and intensive care are crucial to successful treatment of this fulminant infection. After poliovirus is eradicated, EV71 will become the most important enterovirus that affects children, and development of a vaccine may be the only effective measure against it. PMID- 11938498 TI - Outbreak of avian influenza A(H5N1) virus infection in Hong Kong in 1997. AB - The first outbreak of avian influenza A(H5N1) virus in humans occurred in Hong Kong in 1997. Infection was confirmed in 18 individuals, 6 of whom died. Infections were acquired by humans directly from chickens, without the involvement of an intermediate host. The outbreak was halted by a territory-wide slaughter of more than 1.5 million chickens at the end of December 1997. The clinical spectrum of H5N1 infection ranges from asymptomatic infection to fatal pneumonitis and multiple organ failure. Reactive hemophagocytic syndrome was the most characteristic pathologic finding and might have contributed to the lymphopenia, liver dysfunction, and abnormal clotting profiles that were observed among patients with severe infection. Rapid diagnosis with the use of reverse transcription polymerase chain reaction and monoclonal antibody-based immunofluorescent assay were of great clinical value in the management of the outbreak. The experience of the H5N1 outbreak in Hong Kong underscores the importance of continuous surveillance of influenza virus strains in humans and in other animal species. PMID- 11938499 TI - Pediatric acquired immunodeficiency syndrome in Asia: mother-to-child transmission. AB - At least 6 million people in the Asia-Pacific region have human immunodeficiency virus (HIV) infection or acquired immunodeficiency syndrome (AIDS). The numbers of HIV-infected women and children are increasing at an alarming rate. Important components of infection control that need to be addressed if mother-to-child transmission of HIV is to be prevented successfully include voluntary and confidential counseling and testing, family planning, obstetric care, use of antiretroviral agents, and availability of alternatives to breast-feeding. Many services can aid in the prevention of mother-to-child transmission of HIV and the care of HIV-infected mothers and their children. Each country needs to take into account the features of HIV and AIDS epidemiology that are peculiar to it, its infrastructure, and the resources that are available. Providing services in a stepwise manner can help prevent mother-to-child transmission of HIV. PMID- 11938500 TI - Changing incidence of primary total hip arthroplasty and total knee arthroplasty for primary osteoarthritis. AB - This study reports on the incidence of primary total hip arthroplasty (THA) and total knee arthroplasty (TKA) for primary osteoarthritis in Australia. Age specific and gender-specific numbers for Australia, 1994 through 1998, and South Australia, 1988 through 1998, were obtained. Incidences were calculated per 100,000 population. In Australia, primary THA increased from 50.9/10(5) (1994) to 60.9/10(5) (1998). TKA increased from 56.4/10(5) to 76.8/10(5). Stratified by age and gender, changes in incidence for South Australia with respect to time were tested using regression analysis. South Australia showed a significant increase in the overall incidence of THA (P=.012) and TKA (P<.001), although this was not uniform across all age groups. No significant gender differences were found. The incidence of THA is increasing, and the incidence of TKA is increasing at a greater rate. PMID- 11938501 TI - Causes of death after total hip arthroplasty: a nationwide cohort study with 24,638 patients. AB - Based on the nationwide registration of the total hip arthroplasties (THAs) in Finland since 1980, a cohort of 24,638 patients with primary THA was gathered and followed for causes of death until December 31, 1996. The causes of death were divided into 20 main categories according to the classification of diseases ICD 10. The number of person-years was 153,410, and the mean length of follow-up of a person was 6.2 years. During the follow-up, 4,626 patients died; the expected number was 6,746. The standardized mortality ratio (SMR) was 0.69 (95% confidence interval; 0.67-0.70), without any difference between men and women. The total risk increased during the follow-up, with the highest being 0.84 (95% confidence interval, 0.81-0.87). Among the ICD categories, there were significantly low SMRs for cancers (0.54), accidents (0.74), cardiovascular diseases (0.70), and respiratory diseases (0.46). Among the diseases, there was a constant and significant decline of the SMR for dementia and Alzheimer's disease (0.50), diabetes (0.40), myocardial infarction (0.73), hypertension (0.68), other ischemic diseases (0.70), other heart diseases (0.57), and cerebrovascular diseases (0.70). The explanation for the decreased SMRs seems to be attributed to factors other than the THA per se, such as preoperative patient selection, more active lifestyle after THA, and possibly the use of anti-inflammatory drugs. PMID- 11938502 TI - Factors predisposing to dislocation after primary total hip arthroplasty: a multivariate analysis. AB - We conducted this study to determine the relative influence of various mechanical and patient-related factors on the incidence of dislocation after primary total hip asthroplasty (THA). Of 2,023 THAs, 21 patients who had at least 1 dislocation were compared with a control group of 21 patients without dislocation, matched for age, gender, pathology, and year of surgery. Implant positioning, seniority of the surgeon, American Society of Anesthesiologists (ASA) score, and diminished motor coordination were recorded. Data analysis included univariate and multivariate methods. The dislocation risk was 6.9 times higher if total anteversion was not between 40 degrees and 60 degrees and 10 times higher in patients with high ASA scores. Surgeons should pay attention to total anteversion (cup and stem) of THA. The ASA score should be part of the preoperative assessment of the dislocation risk. PMID- 11938503 TI - Independent contribution of elevated-rim acetabular liner and femoral head size to the stability of total hip implants. AB - The use of an elevated-rim acetabular liner in total hip arthroplasty is widely accepted. We sought to determine quantitatively the amount of additional stability provided by the elevated-rim liner compared with the standard nonelevated liner. The stability of the hip with a 32-mm femoral head was compared with the standard 28-mm head. Our results show that a 15 degrees elevated-rim acetabular liner placed in the posterior quadrant increased hip stability by an additional 8.9 degrees of internal rotation. Similarly the 32-mm head provided 8.1 degrees of additional internal rotation. The increases were statistically significant (P<.0001). We did not encounter increased anterior dislocation intraoperatively. The findings of this study indicate that the 32-mm head may contribute to hip stability in primary total hip arthroplasty, and in instances in which a posterior approach is used, an elevated-rim liner placed in the posterior quadrant may contribute independently to hip stability. PMID- 11938504 TI - Unicompartmental knee arthroplasty: 3- to 10-year results in a community hospital setting. AB - The results of 40 medial unicompartmental knee arthroplasties, performed by a single surgeon in a community hospital setting, with an average follow-up of 6 years (range, 3-10 years), were analyzed retrospectively. The Knee Society rating system was used, revealing an average score of 80.5 points and 79% good-to excellent results. One patient required revision to total knee arthroplasty secondary to late infection. Five other patients (5 knees) required further surgery consisting of limited synovectomy and exchange of polyethylene insert at an average of 5.5 years. The 10-year survival rate using revision to total knee arthroplasty or aseptic loosening as an endpoint is 97%. The survival using any revision surgery as an endpoint was 74% at 10 years. Statistical analysis revealed that the most important factor for good-to-excellent results was patient age >65 years. Likewise, postoperative alignment was crucial in decreasing the rate of revision surgery. No other selection criteria were statistically significant in this series. We report favorable intermediate-term results in 40 unicompartmental knee arthroplasties; special attention to patient selection and postoperative alignment is essential to attain good-to-excellent results. PMID- 11938505 TI - Perioperative blood salvage as an alternative to predonating blood for primary total knee and hip arthroplasty. AB - A total of 200 consecutive patients who underwent primary total knee or hip arthroplasty were reviewed to assess the efficacy of perioperative blood salvage and retransfusion. Five of 132 (3.8%) patients undergoing total knee arthroplasty and 3 of 68 (4.4%) patients undergoing total hip arthroplasty required allogeneic transfusion in addition to retransfusion of salvaged autologous blood. The risk of receiving allogeneic transfusion in addition to retransfusion of salvaged blood was 1.2% (2 of 173) in patients with a preoperative hematocrit of > or=37%. The risk of requiring allogeneic transfusion was 22% (6 of 27) in patients with a preoperative hematocrit of 37%. PMID- 11938506 TI - The effect of substrate roughness and hydroxyapatite coating thickness on implant shear strength. AB - This study examined the role of substrate preparation and hydroxyapatite (HA) coating thickness on bone ongrowth and shear strength in a bilateral bicortical sheep model. Plasma-sprayed and grit-blasted titanium implants with different thickness HA coatings were examined at 4, 8, 12, and 26 weeks after implantation. Shear strength increased with time for all implants. Plasma-sprayed implants were superior to grit-blasted implants at all time points. The 100-microm-thick HA layer used in the present study provided greater fixation and ongrowth and less resorption compared with the 50-microm-thick layer. We did not observe any advantage in using a thicker HA coating for the titanium substrates examined. PMID- 11938507 TI - The effect of the ankle brachial pressure index and the use of a tourniquet upon the outcome of total knee replacement. AB - Current teaching suggests that patients requiring a total knee arthroplasty who have absent pulses or an abnormal ankle-brachial pressure index (ABPI) should be referred for vascular investigation. The aim of this study was to determine whether total knee arthroplasty was safe in such patients. The ABPI was measured with a hand-held Doppler. Patients were excluded if they had active ulceration, rest pain, or an absent femoral pulse, but patients with claudication were included. A total of 73 patients were recruited. Hand-held Doppler detected signals from 98% of arteries. Postoperatively, there were no signs of limb ischemia, and the median ABPIs increased significantly. From our series, it seems safe to proceed with surgery in patients with impalpable foot pulses or claudication, as long as the femoral pulse is palpable and there is no active ulceration or rest pain. PMID- 11938508 TI - Does body weight influence outcome after total knee arthroplasty? A 1-year analysis. AB - Obesity has been considered an adverse influence on the outcome of total knee arthroplasty (TKA), and overweight patients often are advised against having a TKA. The role of body weight has not been shown conclusively using objective outcome measures. A group of 180 patients undergoing primary TKA performed by a single surgeon for osteoarthritis was studied prospectively. Their baseline health status and knee scores were recorded, and changes in these measures were studied at 3 and 12 months after surgery. The Nottingham Health Profile was used to assess health status, and the Knee Society score was used to assess clinical outcome. Body mass index was used as a measure of obesity. We found that body weight did not influence adversely the outcome of TKA in the short-term. PMID- 11938509 TI - The use of a carbon fiber plate for periprosthetic supracondylar femoral fractures. AB - Operative management of periprosthetic supracondylar femoral fractures in poor quality bone presents a challenging task for the orthopaedic surgeon. We report our experience with the use of a semirigid carbon fiber-reinforced plate in the treatment of 5 patients, all of whom were elderly women with severe osteoporosis and highly restricted mobility. All 5 of the fractures were sustained after low energy trauma at least 2 years after primary knee arthroplasty. Except for 1 patient who died of pulmonary embolism, all 4 surviving patients progressed uneventfully to fracture union with good mobility and no residual pain. The use of this method for the treatment of periprosthetic femoral fractures has not been reported previously, and we believe it provides a successful and technically undemanding solution to this difficult problem. PMID- 11938510 TI - Impacted corticocancellous allografts and cement for femoral revision of total hip arthroplasty using Lubinus and Charnley prostheses. AB - Femoral revision of total hip arthroplasty using impacted morcellized allografts and a cemented Lubinus SP-II prosthesis (Waldemar Link GmbH & Co, Hamburg, Germany) or Charnley standard prosthesis (De Puy, Leeds, UK) was performed in patients with loss of bone stock grade II-IV according to the Endoclinic classification from first to eighth revision. We report the results from 57 hips in 56 patients, with a median of 64 months' follow-up. Modified Merle d'Aubigne Postel postoperative scores increased significantly from preoperative to postoperative values for the Lubinus group and for the Charnley group. Few complications were seen in this follow-up period. Five stems had minor subsidence. Mechanical failures were 4% for both groups. Radiologic examinations provided evidence of frequent trabecular remodeling of the graft, especially for the Lubinus prosthesis. PMID- 11938511 TI - Exposure to particles stimulates superoxide production by human THP-1 macrophages and avian HD-11EM osteoclasts activated by tumor necrosis factor-alpha and PMA. AB - Wear of orthopaedic implants generates particles capable of inducing bone resorption and aseptic loosening of the implant. The present study shows the combined effect of particles and cell activation on macrophage (THP-1) and osteoclast (HD-11EM) release of reactive oxygen and nitrogen species, providing insight into mechanisms that can lead to osteolysis. In the absence of cell activation, exposure of either cell type to submicron zirconia or latex particles did not elicit an increase in reactive oxygen and nitrogen species production. Suboptimal stimulation with 4 beta-phorbol-12-myristate-13-acetate (PMA) plus particles resulted in a synergistic release of superoxide (O2-), however, and a low-level production of nitric oxide small middle dot by THP-1 macrophages. Similarly, particle stimulation of tumor necrosis factor-alpha-activated THP-1 cells increased O2- release. Our findings show the synergistic effect of cell activation and wear particles on O2- production by activated macrophages and osteoclasts, suggesting O2- involvement in mediating osteolysis. PMID- 11938512 TI - Does the lumbar spine need to be supported to stabilize the pelvis during total hip arthroplasty in the lateral position? AB - Total hip arthroplasty is done commonly with the patient in the lateral position. It has been postulated that the lumbar spine may sag in this position because it forms a mobile link between the rigidly held thoracic spine and the sacrum and may cause the pelvis to tilt in the coronal plane, leading to malpositioning of the acetabular cup. To confirm this hypothesis, 10 consecutive patients seen in the preoperative assessment clinic had 2 anteroposterior radiographs taken in the lateral position. The area of exposure was from the lower thoracic spine to the sacrum. In the first radiograph, the lumbar spine was unsupported, and in the second, it was supported. The radiographs showed that the support did not make any significant difference to the position of the lumbar spine or to the position of the pelvis. PMID- 11938513 TI - Impact biomechanics and pelvic deformation during insertion of press-fit acetabular cups. AB - Five fresh cadaver pelves were cleaned of soft tissue and instrumented with strain gauges. The acetabula were reamed, and a cementless cup, oversized at the periphery, was inserted. The applied force and cup acceleration were measured during insertion and used to calculate an effective mass of the cup, insertion device, and pelvis during each impact. Periacetabular strains increased variably during cup seating. After the cups were seated, strains continued to increase with postseating impacts. The effective mass remained constant throughout the test, indicating that cup seating is not associated with a change in acceleration. This finding implies that an accurate assessment of cup seating cannot be inferred by surgeon proprioception during impaction, and use of an apical hole in the cup is necessary to determine when the cup has seated. PMID- 11938514 TI - The clinical significance of femoral head culture results in donors after hip arthroplasty: a preliminary report. AB - The aim of this study is to determine whether a link exists between clinical wound problems and positive bacterial culture of the femoral head after primary hip arthroplasty. Clinical data were retrieved for 24 culture-positive and 26 culture-negative patients. Coagulase-negative Staphylococcus was the commonest pathogen, cultured in 19 of the 24 culture-positive femoral heads donated to a regional bone bank. Wound problems occurred in 2 of the patients from the culture positive group and 4 of the patients from the culture-negative group. The relative risk of wound problems was 0.54 in the culture-positive group. Based on the available evidence in this preliminary short-term retrospective review, positive microbiology from donated femoral heads has no clinical implication in the donors. The practice of reporting culture-positive femoral heads should be continued. PMID- 11938515 TI - Comparison of a mechanical acetabular alignment guide with computer placement of the socket. AB - We hypothesized that use of mechanical acetabular guides for intraoperative alignment leads to variations between the actual and desired implant orientation. Acetabular implant orientation using only the mechanical guide was studied in 78 patients (82 hips) undergoing primary total hip arthroplasty. A computer-assisted navigation system was used to measure alignment and to monitor the orientation of the pelvis during surgery. When using the mechanical guide, there was significant variation in cup alignment from the desired goal of 45 degrees of abduction and 20 degrees of flexion, and this would have resulted in unacceptable acetabular alignment in 78% of hips. With the support system used, there was significant variability in pelvic orientation during surgery. The mean anteversion of the pelvis was an average of 18 degrees from the optimal orientation. These results show a clear need to develop more reliable tools than were used or anatomically based alignment strategies to provide reproducible and accurate acetabular alignment. PMID- 11938516 TI - Supracondylar fractures of the femur above total knee arthroplasties with cortical allograft struts. AB - Ten supracondylar fractures of the femur above a total knee arthroplasty were treated with open reduction and internal fixation using a metal plate and cortical allograft struts. Failure with nonunion of previous operative treatments was seen in 5 patients. The other 5 patients had either severe osteopenia or comminution of the fracture above the knee prosthesis. At an average of 38 months' follow-up, all the fractures healed with an average union time of 17.6 weeks. The postoperative knee alignment averaged 5.1 degrees valgus. Infection occurred in 1 patient, which resulted in a poor outcome. The remaining 9 patients were functioning well at the most recent follow-up examination. The use of cortical allograft struts combined with a compression plate provided satisfactory results for supracondylar fractures above knee prostheses with severe osteopenia or loss of bone stock after initial failure of internal fixation. PMID- 11938517 TI - Disseminated osteosarcoma arising in the pelvis after total hip arthroplasty. AB - The development locally of a malignant neoplasm after total hip arthroplasty is rare but increasingly recognized. Its significance is unclear. The association could be fortuitous. If not, the rarity of the complication argues that constitutional factors are likely to be important in pathogenesis. We describe a case of disseminated osteosarcoma of the hip developing 6.5 years after total hip arthroplasty and review the related literature. PMID- 11938518 TI - Delayed transient sciatic nerve palsy after total hip arthroplasty. AB - Sciatic nerve palsy after total hip arthroplasty is a well-known complication, but delayed sciatic nerve palsy is rare. We report such a case with profound clinical manifestations and well-documented electrophysiologic changes. We found no helpful guidance to managing delayed palsy in the literature. We also are unaware of any previous cases reported in which nearly full recovery has occurred. PMID- 11938519 TI - Migration of intrapelvic cement after total hip arthroplasty. AB - Intrapelvic extrusion of cement during total hip arthroplasty is a frequent occurrence. We report a case in which the intrapelvic cement mass broke free 3 years after the primary procedure and migrated proximally to lie against the posterior abdominal wall, resulting in intractable groin pain. The patient was relieved of pain after removal of the cement mass. PMID- 11938520 TI - Brucellosis as a cause of septic loosening of total hip arthroplasty. AB - Infection after a total hip arthroplasty is a severe complication and is associated with a high incidence of morbidity. We describe a case of late infection, 5 years after the implantation of a cemented hip prosthesis. The infection was caused by gram-negative Brucella melitensis and occurred in a 63 year-old man who owned cattle. As far as we know, such a complication has never been published before. PMID- 11938521 TI - [Recommendations for clinical practice. Perioperative treatment in endonasal surgery]. PMID- 11938523 TI - [Mucous malignant melanoma of the neck and face. Experience at the Salah Azaiz Institute (Tunis)]. AB - The mucosal melanoma of the head and neck is rare and of late diagnosis. This retrospective study concerns 17 cases brought together in 30 years. Sex-ratio was of 1.1, the average age of 58 years. Tumoral seats were the following ones: nasal cavity and paranasal sinus (n = 10), nasopharyngeal cavity (n = 2), gingival seat (n = 2), palate (n = 1), laryngeal seat (n = 1), middle ear (n = 1). Tumoral extension was classified an follows: stage I: 52.9%, stage II: 17.6%, stage III: 29.4%. Melanomas were achromic in 23.5% of cases. On the therapeutic plan, 47.1% of the patients were treated in a purpose palliative because of the importance of the tumoral extension. Seven patients (41.2%) were treated surgically; three among them received an additional radiotherapy for an insufficient tumoral excision or adenopathy in break capsular. Two patients (11.8%) were treated by exclusive radiotherapy. The rates of survival were 17.6% in 2 years, and 5.9% in 5 years. The average duration of survival was of 18.1 months. The causes of failure were essentially local and metastatic. PMID- 11938522 TI - [French multicenter prospective epidemiologic study (ORLI Group) of allergic and lung diseases associated with nasal polyposis]. AB - 224 patients presenting with nasal polyposis (NP) were included in a french prospective multicenter study. NP was evaluated by nasal endoscopy and computed tomography. Allergic status was documented using skin prick-tests and/or specific IgE. Pneumologic assessment included spirometry with carbamyl-choline hyper reactivity test or beta 2 mimetic broncho-dilation test. Minimal follow up period was 1 year. 45% of the whole population were considered as asthmatic. Asthma onset occurred before and after the NP onset in respectively 45.7%, 22.3% and 32% of cases; these two conditions started simultaneously in 32% of patients. Skin prick-tests and/or specific IgE were positive in 32.5% of cases. In most of the cases (80%), patients were polysensitized to house dust mite and/or pollens and/or animal danders and/or fungi. 31% of the population had idiosyncrasy, caused by drugs in general and especially aspirin in 44% of cases. The global population could be divided in two groups according to the occurrence of previous polypectomy or not. The group "polypectomy" and the group "no polypectomy" were similar regarding the frequency, the age of onset, the course and the severity of associated asthma. Familial history (parents, children, brothers and sisters) was of great interest: 58.7% of the patients had one (or more) relative suffering from NP, 43.6% of the patients had one (or more) relative suffering from asthma and 12.2% of the patients had one (or more) relative suffering from idiosyncrasy. These results support a genetic etiology for NP. PMID- 11938524 TI - [Loss of visual acuity caused by ethmoidal mucocele cured after endonasal surgery: report of a case]. AB - A posterior and/or ethmoidal mucocele remains a rare occurrence. The condition usually follows a past history of sinonasal surgery. Extension of the lesion may have ophtalmological implications, with the possibility of causing visual impairment. The predilection for a site of origin in the posterior sinuses explains the frequent absence of nasal symptoms, and the usual pattern of discovery by the ophtalmologist. Modern imaging techniques, especially scanning, allows the diagnosis to be made immediately after the first orbital symptoms. By the same token, recent techniques of endoscopic endonasal surgery have radically changed the treatment. Early complete surgical removal holds out hope for remission of the symptoms that led originally to discovery of the mucocele. We report here a case of posterior ethmoidal mucocele causing a visual field defect which reversed completely after endoscopic treatment. PMID- 11938525 TI - [Unusual benign tumors of the larynx: report of 17 cases]. AB - Rare benign tumours of the larynx (RBTL) form a heterogeneous group. The clinical picture is non-specific. Based on a series of 17 cases of RBTL treated in our department, we wish to draw attention to the diagnosis, treatment and clinical course of these tumours. The mean age of the patients was 37 years, with the male sex predominating. The presenting symptom was most commonly progressive dysphonia, while 5 patients were admitted in respiratory distress. Examination and special investigations revealed 8 cases of laryngocoele, 6 cases of angioma, 2 cases of schwannoma, and one of chondroma. Treatment was surgical, by an external approach in 10 cases and endoscopically in 7. The postoperative course was satisfactory. No long-term recurrences were encountered. Conservative surgery by an external or endoscopic approach has revolutionised the functional outcome in these benign tumours. PMID- 11938526 TI - Malignant mixed tumor of the larynx: a case report. AB - Malignant mixed tumour (MMT), which is a salivary gland tumour, is the malignant form of pleomorphic adenoma. Although the tumour can also originate from the minor salivary glands throughout the submucosa in the head and neck region, laryngeal involvement is quite rare. An additional case of laryngeal MMT presented in a forty-five year old man, and the diagnostic, immunohistochemical (S-100, actin, vimentin, epithelial membrane antigen) and therapeutic procedures were presented. PMID- 11938527 TI - [Value of positron-emission tomography in the post-treatment follow-up of epidermoid carcinoma of the head and neck]. AB - This study aimed at pointing out the supply of the positron emission tomography (PET) in the posttherapeutic follow-up of the head and neck squamous cell carcinomas and to determine the best period to perform this test. PATIENTS AND METHODS: Twenty patients have been included in this series, 16 men and 4 women. The PET was performed between 3 and 6 months after the end of all therapy. It systematically included radiation therapy. The results of the PET have been compared with those obtained by histology. The average distance of the follow-up of the patients after the achievement of the test was 11 months. RESULTS: They divided up according to the presence or not of an abnormal fixation on the PET imaging. Negative PETs: eight cases. Among those, a patient showed a metastatic cervical adenopathy at five months. Positive PETs: twelve cases which can be divided into three groups according to the area of the fixation. Primary site: 8 cases, 4 of which false-positive. Cervical lymph nodes: one case. Other sites: three cases. In our series PET had a sensitivity of 87% and a specificity of 67%. CONCLUSION: The PET is an original imaging as it allows a corporal metabolic study at one go. It seems to be very useful in the follow-up of patients who show a head and neck squamous cell carcinoma. The best period to perform it is the third or fourth posttherapeutic month. The high sensitivity is interesting within the context of an early detection of a residual tumour, for it allows to think of a suitable therapy quicker. PMID- 11938528 TI - Extramuscular soft tissue myxoma of the lateral neck. AB - Myxomas are rare benign connective tissue tumours of unclear histogenesis. In this case a lateral neck tumour initially behaved, both clinically and radiographically as a lipoma, and was therefore managed conservatively. Subsequently the lesion caused pressure symptoms and therefore a surgical approach was taken with subsequent histology confirming a myxoma. Only five cases of soft tissue myxoma of the lateral neck have been reported in the English literature of which two were extramuscular. This is the largest reported myxoma of the head and neck region and was treated with simple enucleation. There has been no evidence of recurrence five years after surgery. PMID- 11938529 TI - A histopathologic evaluation of mucosa obtained from mastoidectomy cavity by mastoidoscopy. AB - Despite the routine application of intact canal wall mastoidectomy for chronic suppurative otitis media, the outcome of mucosa within the mastoidectomy cavity is not wellknown. The purpose of this was to evaluate the mastoidectomy cavity and obtain mucosal study specimens from the mastoidectomy cavities in order to reveal any residual pathology. Mastoidoscopy under local anaesthesia was performed to evaluate the mastoidectomy cavity and obtain mucosal specimens from the mastoid antrum. The procedure was carried out in 12 patients. Four (33%) patients had wide cavities, 6 (50%) had their cavities narrowed down to the antrum, and 2 (17%) patients had obliterated cavities. Mucosal specimens examined with transmission electron microscopy revealed findings like swelling of some of the mitochondria, decrease in the number of organelles, intracellular oedema, changes in the nuclei, decrease in cilia and fibrosis. In conclusion, even if there may be some pathology at a microscopic level, this does not necessarily lead to any clinical problem. PMID- 11938530 TI - [Ambulatory surgery of otosclerosis: retrospective study of 102 cases]. AB - A retrospective study of 102 selected patients operated for otosclerosis (34 outpatient surgery, 68 hospitalised), having all of them the inclusion criteria of the ambulatory surgery, treated as outpatient in a traditional health sector or hospitalised, depending on their own choice, has been lead. We analysed the results of the pure tone audiometric tests two to six months after surgery. No significative difference was found between the two groups on audiometric results as for the postoperative complications. On the other side, it seems that young patients are more interested by the one-day surgery. The failure of the ambulatory surgery could be explained by the vertigo or dizziness per- or postoperatively. Finally, the evaluation of the cost-benefit shows that the ambulatory surgery in a traditional health sector could lead a budgetary saving policy. A saving way that will grow in a specialized sector devoted to the ambulatory practice. PMID- 11938531 TI - [The child as bone marrow donor]. PMID- 11938532 TI - [Heterozygote state mutations of the growth hormone receptor gene: what is its significance?]. PMID- 11938533 TI - [HIV exposure accidents in children. Experience at the emergency admission services of the CHU of Nice]. AB - Accidental exposure to human immunodeficiency virus (HIV), either by injury with a discarded syringe or after sexual abuse, increases among the children population. PATIENTS AND METHODS: The entire period of the study was seven-year. Twenty-eight children (14 males, 14 females) underwent three visits after an accidental exposure at Day 0, Day 30th and Day 90th. During each visit, a physical examination, liver enzymes, Ag P24, PCR-RNA HIV, B, C hepatitis serologies were performed. An antiretroviral prophylaxis was proposed when a high risk of HIV transmission was recognized. Anti hepatitis B globulins were administrated when no previous immunization had been made. RESULTS: The median age was 9.3 years (range: 1.5 to 16.7 years). 93% of the children consulted within 48 hours after exposure. Five of them consulted after a sexual aggression with penetration, two after a mucosus exposition and two after deep needle injuries. The source of the contamination was known in five cases. Nine patients were given an antiretroviral treatment for four weeks (seven bitherapy, two tritherapy), with no major side effects. Eighteen and 11 children were seen at the second and third visits, respectively. No case of HIV or HCV infection was observed. CONCLUSION: No case of infection after injury with a discarded syringe was found in our study. This situation differs from what is observed in health care workers, where the higher risk factor is observed. Even if no contamination was found after sexual aggression in our population, the risk of HIV is actual. This is why we propose an antiretroviral prophylaxis against HIV in only one indication, corresponding to sexual penetration. PMID- 11938534 TI - [Use of a polyethylene bag: a way to improve the thermal environment of the premature newborn at the delivery room]. AB - BACKGROUND: Early interventions, such as occlusive wrapping of very low birth weight infants at delivery reduce postnatal temperature fall. This new intervention was implemented in our hospital on January 2000. The aim of this study was to investigate retrospectively the effect of polyethylene wrap, applied immediately at birth, on thermoregulation. PATIENTS AND METHODS: Matched pair analysis was conducted for 60 infants delivered inborn at less than 33 weeks' gestation and 60 premature infants who were born during the second half of 1999 fulfilling the same criteria. The only difference in the management (medical and environmental) was wrapping with a polyethylene bag in the delivery room. Rectal temperature and other vital parameters were taken, after removal of wraps, on admission to NICU. RESULTS: The perinatal characteristics of both groups were comparable. Use of wrapping resulted in a significantly higher admission rectal temperature (difference in means = 0.8 degree C, p < 0.0001), this difference was also significant in infants < 30 weeks. The incidence of hypothermia (< 35.5 degrees C) was less frequent in infants enclosed in plastic bags (8.3% vs 55%). No side effects (skin burns, infection or hyperthermia) were attributable to the intervention. The heart rate was higher in the wrapping group (163 +/- 16 vs 150 +/- 17 b/min, p < 0.01), as well as the capillary glycemia (62 +/- 26 vs 45 +/- 30 mg/dl, p < 0.01). There was no significant difference on arterial pressure. CONCLUSION: Occlusive wrapping with a polyethylene bag at birth prevented low rectal temperature in premature infants in the immediate postnatal period. This method is easy, practical and effective, and does not interfere with current practice for resuscitation. PMID- 11938535 TI - [Detection of exercise-induced bronchospasm in sixth-grade students: impact of health education]. AB - Underdiagnosis of asthma is frequent in children and may be improved by the development of school-based health programs. MATERIAL AND METHODS: We developed an educational program in 11-year-old schoolchildren who participated in a screening test for exercise-induced asthma (EIA). All children were given an asthma questionnaire before and after two educational sessions. RESULTS: Mean score for asthma knowledge quiz increased from 63 to 85% (p < 0.001). Thirty-six children (3.7%) were initially considered as asthmatic and showed better responses than non-asthmatic children. Peak expiratory flow (PEF) was measured before and after an outside running exercise. A fall in PEF of at least 15% was considered an abnormal result. Seventy-height children (8%) had a decrease in PEF, including 65 children initially not recognized as asthmatic. For these latter, a medical evaluation was recommended to parents. Only 28 of these children gave informations on follow-up: 19 had lung function tests, two received a treatment without preliminary function test, and seven had neither lung function test nor treatment. Among the children who realized lung function tests, five had criteria for airway obstruction at baseline, and ten had significant bronchial hyperreactivity. CONCLUSIONS: However, a new screening test revealed that only a small minority of children initially not recognized as asthmatic but having decreased their PEF, were consequently considered as asthmatic by their practitioner, even in case of positive lung function test. PMID- 11938536 TI - [Hospitalization for acute community-acquired rotavirus gastroenteritis: a 4-year survey]. AB - OBJECTIVE: To evaluate the number of hospitalizations due to community-acquired rotavirus acute gastroenteritis in a general pediatric unit during a four-year survey. RESULTS: From January 1997 to December 2000, 725 patients were admitted for acute gastro-enteritis to the general paediatric unit of a Parisian children hospital (nosocomial diarrhoea excluded) and 706 (97.5%) of these patients had had a stool microbiologic examination. Diarrhoea was caused by rotavirus in 359 patients (50.89%) and Salmonella sp in 61 (8.6%). Children and infants hospitalized for rotavirus acute gastroenteritis were younger (26% had three months or less, and 50.03% had six months or less) than in other European studies. CONCLUSION: This study is the first in France reporting a systematic survey of hospitalized gastroenteritis during four years. More than half of hospitalized community-acquired gastroenteritis were due to rotavirus in this Parisian area. The young age of patients should be investigated in other French areas, searching for risk factors and rotavirus strains. PMID- 11938537 TI - [Varicella complicated with necrotizing fasciitis caused by group A hemolytic Streptococcus]. AB - Chickenpox has a high risk of invasive group A streptococcal disease and necroziting fasciitis. CASE REPORT: A five-year-old girl, during chickenpox treated with ibuprofen, developed sepsis and edematous and necrotic lesions of the pelvis and the abdominal wall. The child improved with surgical treatment and adjunction of clindamycin to the antibiotic therapy. CONCLUSION: We review the optimal medical and surgical treatment of necrotizing fasciitis and discuss the role of chickenpox and non steroidal antiinflammatory agents in this disease. PMID- 11938538 TI - [Central alveolar hypoventilation syndrome and cerebral venous thrombosis: fortuitous association?]. AB - Central hypoventilation syndrome is defined as the failure of automatic control of breathing. Secondary central hypoventilation syndrome should distinguish from congenital central hypoventilation syndrome by brainstem abnormalities, place of respiratory control. CASE REPORTS: We report two clinical cases characterized by late onset central hypoventilation syndrome (three years--six months, and five years old): in the first case the diagnosis was made after general anesthesia and the second one presented with acute nocturnal comatose state. Neuroradiologic investigations showed bilateral cerebral sinus veinous thrombosis without any brainstem lesions. Moreover these children had severe behavior disorders: psychomotor instability, alterations of social relations, language dysfunction, and neurocognitive deficit. This symptomatology seems independent from central hypoventilation syndrome and cerebral venous thrombosis. CONCLUSION: Late onset central hypoventilation syndrome may be associated with cerebral venous thrombosis. Ischemia of central chemoreceptors or integration of their informations could be one of mechanism. PMID- 11938539 TI - [Extracorporeal oxygenation in newborn infant born to diabetic mother with severe hypertrophic myocardiopathy]. AB - Hypertrophic cardiomyopathy is a common complication in infants born to diabetic mothers but it is rarely life threatening. CASE REPORT: A neonate born to a diabetic mother with a severe hypertrophic cardiomyopathy with life threatening respiratory and haemodynamic distress was successfully supported by extracorporeal membrane oxygenation. CONCLUSION: Extracorporeal membrane oxygenation can be an efficient transitory therapy for very severe hypertrophic cardiomyopathy in infants of diabetic mothers. PMID- 11938540 TI - [Adrenal hematoma associated with rheumatoid purpura]. AB - Schonlein-Henoch purpura is a systemic vasculitis involving primarily the skin, musculoskeletal system, gastro-intestinal tract, and kidneys. The purpose of this paper was to report the first described pediatric case, to our knowledge, of adrenal hemorrhage occurring in Schonlein-Henoch purpura. CASE REPORT: Our patient was a 12 year-old girl who presented a right adrenal hematoma diagnosed following an episode of acute right flank pain, ten days prior to the appearance of the clinical symptoms of Schonlein-Henoch purpura. CONCLUSION: We think that Schonlein-Henoch purpura should be included in the diagnosis of adrenal hematomas. PMID- 11938541 TI - [Community-acquired pneumonia in children: etiology and treatment]. AB - Due to their very different etiological agents, community-acquired pneumoniae in children frequently require empiric antibiotic therapy in emergency. Streptococcus pneumoniae represents between 15 to 30% of the etiologies and has unspecific diagnostic procedures; as a matter of fact radiological lobar consolidation is seen in less than half of cases, and laboratory data, except for high procalcitonin level, are poorly reliable. Pneumonia due to Mycoplasma pneumoniae is frequent after 2 years of age, reaching 40 to 60% of causes in ambulatory teenagers; it must be treated with macrolides as sequellae are possible. The exact number of viral pneumonia is difficult to establish because of the lack of reliable diagnostic methods. If bacterial superinfections are probably overestimated during acute phase, viral infections may lead to bacterial pneumonia 2 to 4 weeks after the initial episode. Empiric antibiotic treatment must take into account pneumococci and their penicillin-resistant strains. Amoxicillin is the antibiotic of choice, having a higher efficacy on resistant pneumococci than oral cephalosporins. In case of clinical failure of amoxicillin, mycoplasma infection must be considered and patient must receive macrolides. Future epidemiology will be affected by anti-pneumococcal immunisation but difficulties in diagnosis and empiric antibiotic treatment will probably remain. Studies in immunised children are needed to evaluate the importance of pneumococcal infections due to serotypes not included in the vaccine. PMID- 11938542 TI - [Contribution of antineoplastic biotherapy in the treatment of leukemia in children]. AB - Improvements in the chemotherapeutic and transplant regimens have had a significant impact in improving survival rates for pediatric leukemia. However, there are still major problems to address including what options are available for patients with chemoresistant disease and what strategies are available to avoid toxicity associated with highly cytotoxic treatment regimens. Gene and immunotherapy protocols hold great promise. Using gene transfer of a marker gene, a number of biologic issues in the therapy of leukemia have been addressed. For example, by gene marking autologous bone marrow grafts it has been possible to demonstrate that infused marrow contributes to relapse in acute and chronic myeloid leukemias. In the allogeneic transplant setting, genetically modified T cells have proven valuable for the prophylaxis and treatment of viral diseases and may have an important role in preventing or treating disease relapse. Gene transfer is also being used to modify tumor function, enhance immunogenicity, and confer drug-resistance to normal hematopoietic stem cells. With the continued scientific advancements in this field, gene therapy will almost certainly have a major impact on the treatment of pediatric leukemia in the future. PMID- 11938543 TI - [Radiologic case of the month]. PMID- 11938544 TI - [Ways to reduce fever: are luke-warm water baths still indicated?]. AB - Tepid sponge bathing to reduce temperature may be an aggression against the febrile child organism leading to reactions that will limit the cooling. During the bath, the decrease of the skin temperature induces not only peripheral vasoconstriction, shivering, and metabolic heat production, but also a discomfort. This discomfort occurs when there is a difference between the thermal informations coming from the body and the central thermoregulator set point. It triggers crying when the child has no way to escape. Considering the little effectiveness (0.3 degree C on average) of the association bath cooling and antipyretic treatment versus antipyretic treatment, and the preservation of the child comfort, bathing in tepid water is not useful in current care of children fever. PMID- 11938545 TI - [Risk medications in case of glucose-6-phosphate dehydrogenase deficiency]. AB - The best known morbid effect of glucosephosphate dehydrogenase (G-6PD) deficiency is hemolysis induced by oxidative drugs. When prescribing drugs for G-6PD deficient subjects, two points should be kept in mind: different genetic variants of G-6PD deficiency entail different susceptibility to the hemolytic risk from drugs; thus a drug found to be safe in some G-6PD deficient subjects may not be equally safe in others; the risk and severity of hemolysis is almost always dose related. The purpose of this paper is to underline the main drugs that cannot be safely administrated to G-6PD deficient subjects. They can be separated in drugs that must be avoided by G-6PD deficient subjects (such as sulphonamides, quinolones, nitrofurantoin), and drugs that do not systematically precipitate hemolysis but must nevertheless be prescribed with caution. PMID- 11938546 TI - [Breastfeeding and culture]. AB - Pediatricians consider that breast-feeding is the most appropriate means of nourishing infants. However, in cultural terms, lactation far outweighs this purely dietary function. The act of breast-feeding is very often regarded as being of prime importance and is accordingly strictly controlled. In human societies the choice, duration, and mode of lactation, are governed by various prohibitions or beliefs. Infant feeding habits are transformed by major social changes, such as women working outside the home, female emancipation or the emergence of a consumer society, although this is not always noticed by physicians and health systems. Rather than being organic in origin, most cases of inadequate milk production are due to social causes. Hence, when promoting breast feeding, in addition to argue on the basis of scientific knowledge, physicians must take into account the changes in the social context. PMID- 11938548 TI - [Treatment of chronic hepatitis C with interferon in children]. PMID- 11938549 TI - [Functional neuroanatomy of the autobiographical memory]. AB - Autobiographical memory refers to events and information about personal life and the self. Within autobiographical memory, many authors make a difference between episodic and semantic components. Study of retrograde amnesia gives information about memory consolidation. According to the "standard model" of consolidation, the medial temporal lobe plays a time-limited role in retrieval memory. Functional neuroanatomy studies of autobiographical memory are very few and many are recent. These studies concern which brain regions are involved in the autobiographical retrieval, episodic or semantic autobiographical memory and consolidation process. Results show that autobiographical retrieval depends on specific brain regions like frontal cortex. Concerning memory consolidation, findings are most consistent with the idea that hippocampal complex is involved in both recent and remote memories. PMID- 11938550 TI - [Neuronal networks and memory: role of the hippocampus]. AB - Learning and memory are related both to cognitive processes and to neurobiological mechanisms. The human pathology focused on the role of the hippocampus and animal experiments have analyzed its implications. The most usually admitted hypothesis is that memories are underlied by distributed specific neural networks defined through the strengthening of certain synapses, under the action of the flow of information during learning. The best candidate for this strengthening of the synapses is a change in synaptic plasticity similar to the artificial phenomenon of long-term potentiation. During memory processes, the hippocampus would play a particular role in information processing (analyzing novelty and significance of the information) and would allow the specification of the neural network, mainly in the cortical territories. We report data in olfactory learning in rats comforting these hypotheses. Considering neurochemistry of memory processes, specific synaptic changes and neuromodulatory processes must be distinguished. We report data about vasopressin illustrating both kinds of mechanisms in the hippocampus. PMID- 11938551 TI - [Neural basis for spatial memory in animals: what do hippocampal neurons tell us?]. AB - Recent studies relying on the recording of neuronal unit activity in freely moving rats show the existence of two populations of neurons signalling the animal's location or head direction: place cells found primarily in the hippocampus and head direction cells found in brain areas anatomically and functionally related to the hippocampus. The properties of these two neuronal populations suggest that their activity strongly depends upon information cues stemming from the spatial environment, and also suggest their involvement in spatial memory. Place cells and head direction cells would jointly participate in a neural network allowing the animal to orient in space and to store spatial locations in memory. This network would also be operating in humans, in particular for encoding specific events in episodic memory. PMID- 11938552 TI - [Cellular and molecular mechanisms of memory]. AB - There has been nearly a century of interest in the idea that information is encoded in the brain as specific spatio-temporal patterns of activity in distributed networks and stored as changes in the efficacy of synaptic connections on neurons that are activated during learning. The discovery and detailed report of the phenomenon generally known as long-term potentiation opened a new chapter in the study of synaptic plasticity in the vertebrate brain, and this form of synaptic plasticity has now become the dominant model in the search for the cellular bases of learning and memory. To date, the key events in the cellular and molecular mechanisms underlying synaptic plasticity are starting to be identified. They require the activation of specific receptors and of several molecular cascades to convert extracellular signals into persistent functional changes in neuronal connectivity. Accumulating evidence suggests that the rapid activation of the genetic machinery is a key mechanism underlying the enduring modification of neural networks required for the laying down of memory. The recent developments in the search for the cellular and molecular mechanisms of memory storage are reviewed. PMID- 11938553 TI - [Oxidative stress and gene expression]. AB - Oxidative stress elicits cellular toxicity implicating various targets such as DNA, RNA, proteins, lipids.... Similarly to other stress conditions, it triggers cellular adaptation and the induction of defense mechanisms. Adaptation to this stress is ubiquitous but it displays cell specificity. Adaptation to stress requires the regulation of a large number of genes. Those encoding anti-oxidant and repair genes are induced while others are repressed. The activity of several transcription factors is altered by oxidative stress. Many transcription factors are inhibited because of the oxidation of cysteine residues in their DNA binding domains. In the case of NFI, a cysteine in the transactivating domain is oxidized which leads to inactivation. The mechanisms of transcription factor activation are often complex, as in the case of cJun. This factor is activated by a kinase signalling cascade. Paradoxically, it is also inhibited by stress since one of its cysteines is oxidized; however, this oxidation is repaired in the nucleus by the ref1-thioredoxine system. A similar mechanism is observed for NF kappa B. Alteration of the redox status is observed in several physiological pathological and toxic conditions. It is also provoked by other cellular stress, in particular endoplasmic reticulum stress, hypoxia, shear stress, osmotic and heat shocks. The actual role of oxidative stress during other stress conditions remains to be elucidated. This widespread implication of oxidative stress could account for its role in a variety of disease such as neurodegenerative diseases, cancer, atherosclerosis, and diabetes. The efficacy of anti-oxidant therapy depends on the actual contribution of oxidative stress to these diseases. PMID- 11938554 TI - [Glucose and redox sensitive transcription factors: consequences on the insulin receptor]. AB - During diabetes an oxidizing stress can be observed, whose origins are multiple. This stress can be regarded as a particular messenger, certainly modifying the transduction of the insulinic signal but also the activity of certain transcription factors. Indeed NF-kappa B activity is increased, while Sp1 activity drops in this context. NF-kappa B activation takes certainly part in vascular remodeling and plays a role in the genesis of the oxidative stress. Hence, NF-kappa B activity precociously increases in the states of insulin resistance before hyperglycemia sets in. Sp1 undergoes a significant fall in activity in the presence of glucose. This activity can be restored by antioxydants in the cell culture medium, Sp1 being necessary to insulin receptor expression, the question arises whether this drop in activity modifies significantly the insulin expression. PMID- 11938555 TI - [Advanced glycation end products (AGEs), free radicals and diabetes]. AB - Nonenzymatic glycation, i.e. binding of monosaccharides to amino groups of proteins, gives rise to complex components called "advanced glycation end products" (AGEs), which alter protein structure and functions, and participate in diabetic long-term complications. Glycation and oxidative stress are closely linked, and are often referred to as "glycoxidation" processes. Experimental data support these interactions. a) All glycation steps generate oxygen free radicals, some of these steps being common with these of lipid peroxidation. b) AGEs bind to membrane receptors such as RAGE, and induce an oxidative stress and a pro inflammatory status. c) Glycated proteins modulate cellular oxidative functions: glycated collagens induce an inappropriate oxidative response in PMNs. d) Products of lipid peroxidation (MDA) bind to proteins and amplify glycoxidation induced damages. Glycoxydation intensity increases in diabetes mellitus, ageing, renal failure and other pathological states with oxidative stress. Therapies aiming at limiting glycoxidation take into account its oxidative part. PMID- 11938557 TI - [Oxygen and the superoxide anion. Modulation of NADPH oxidase?]. AB - Oxidative stress which results from an imbalance between oxidant production and antioxidant defense mechanisms can promote modifications of lipids, proteins and nucleic acids. This review focuses on the different pathways leading to Reactive Oxygen Species (ROS) production in particular on NADPH oxidase activation. This enzyme is localized in numerous cells including phagocytes and vascular cells and composed of membrane and cytosolic sub-units. The activation of the NADPH oxidase is largely involved in inflammation associated diseases such as asthma, Systemic Inflammatory Response Syndrome and aging associated diseases such as atherosclerosis and neurodeneratives diseases. The modulation of NADPH oxidase could be a way to limit or prevent the development of these diseases. PMID- 11938556 TI - [Antioxidant and anti-AGE therapeutics: evaluation and perspectives]. AB - Diabetic patients exhibit an oxidative stress status, that is an imbalance between reactive oxygen species and antioxidant defences, in favour of the first ones. This oxidative stress, together with formation of advanced glycation endproducts (AGEs), is involved in diabetic complications. It could thus be of great interest to propose antioxidant and/or anti-AGE therapeutics as complementary treatment in these patients. Antioxidants can be classical molecules such as vitamin E, lipoic acid or N-acetylcysteine. Thus, vitamin E supplementation can improve insulin efficiency and glycemic equilibrium, as shown by the decrease of glycaemia, glycated haemoglobin and fructosamine values. In addition, this kind of supplementation lowers plasma lipid peroxidation and oxidizability of low density lipoproteins, which is involved in the atherogenesis process. Moreover, it allows to fight against complications such as retinopathy. A second category is represented by molecules able to fight against the effects of glycation end-products (AGEs). They can act: either by preventing cellular action of AGEs; this is obtained with soluble receptors of advanced glycation endproducts (sRAGE); or by inhibiting AGE formation (scavenging of reactive carbonyl intermediates). Nucleophilic compounds such as pyridoxamine, tenilsetam, 2,3-diaminophenazone, OPB-9195 or aminoguanidine can act in this way. Aminoguanidine is able to limit the development of the main diabetes-associated complications in animals. A double-blind clinical assay has been conducted in type 2 diabetic patients in the United States and the Canada, in order to determine if aminoguanidine is able to slow down the progression of diabetes induced nephropathy. We will discuss about another guanidic molecule, i.e. metformin, which is also able to scavenge AGEs, in the last part of this review. A third category of molecules is constituted by oral antidiabetic molecules exhibiting antioxidant properties. They are thiazolidinediones (troglitazone) and sulfonylureas (gliclazide). Troglitazone and gliclazide can thus decrease LDL oxidizability and monocyte adhesion to endothelial cells, which is an early step in the atherogenesis process and which is stimulated by oxidised LDLs. Finally, a prospective way is devoted to oral antidiabetic drugs exhibiting both antioxidant and anti-AGE properties. A very used antidiabetic drug of interest is metformin (dimethylbiguanide), since it can prevent diabetes complications not only by lowering glycaemia, but also by inhibiting AGE formation and by stimulating antioxidant defences. The latter therapeutic approach constitutes a future way in the diabetes area, in order both to obtain a better glycemic control and a least development of diabetic complications. PMID- 11938558 TI - [The contribution of invertebrate study to the biology of nitric oxide]. AB - After the identification of nitric oxide (NO) with the endothelium derived relaxing factor, many signaling mechanisms involving NO were identified through experiments on Mammals. NO activates soluble guanylyl cyclase leading to the formation of cGMP, stimulates the ADP-ribosylation of GAPDH, altering the cell energy production and combines with superoxide, generating cytotoxic peroxynitrite. NO was then progressively established as a major messenger molecule in Mammals. It is implied in the regulation of blood vessel dilatation, immune function, development and neurotransmission in brain and peripheral nervous system. Later, parallel findings were observed in Invertebrates and then, NO appeared as a signaling molecule widely spread throughout the animal kingdom and whose functions were highly conserved during evolution. The purpose of this short review is to highlight the contribution of Invertebrate studies to the knowledge of NO biology. PMID- 11938559 TI - [Local exercise and oxidative stress in chronic obstructive broncho pneumopathies: preliminary results]. AB - The role of altered peripheral muscle function in exercise intolerance of chronic obstructive pulmonary disease (COPD) is now well established. However, the mechanisms underlying this phenomen, have not been determined. One hypothesis is that the oxidative stress, that leads to tissue injury may be involved. A recent study has shown that general exercise caused systemic oxidative stress in COPD patients. However, the origin of this stress was not absolutely clear: airways, muscle, both, or other? The aim of this study was first to determine with a systemic approach, whether systemic oxidative stress occur in patients who perform local exercise and then with a muscular needle biopsy approach, to confirm the muscular origin of this oxidative stress. METHODS: In each approach, 7 COPD patients moderate to severe and 7 age-matched subjects performed an endurance test consisting of dynamic strength of the quadriceps against 40% (systemic approach) or 30% (biopsy approach) of maximal voluntary strength at an imposed regular pace until exhaustion. RESULTS: The results showed in each approach, that endurance test duration was significantly decreased in the COPD patients (p < 0.05). In systemic approach, the results showed that blood vitamin E at rest was significantly decreased in the COPD (p < 0.001), with a significant increase in superoxide anion release by stimulated phagocytes (p < 0.001). Local exercise induced, only in COPD, a significant increase in serum MDA (p < 0.05), which is an index of oxidative stress. In the biopsy approach, the results showed that local exercise induced in COPD an increase in muscular levels of MDA. A significant increase in muscular peroxidase glutathion activity (antioxidant) occurred after exercise only in normal subjects (p < 0.05). In conclusion, this study in COPD, confirms the altered peripheral muscle function, reveals a deficit in blood vitamin E and suggest that local muscular exercise causes a muscular oxidative stress in these patients. Further studies are needed to confirm these results and evaluate the implication of this oxidative stress in the myopathy of COPD. PMID- 11938560 TI - [Mechanical behavior of fibroblasts included in collagen lattices]. AB - Striae distensae are characterized by linear, smooth bands of atrophic-appearing skin. Excessive steroid activity, genetic and mechanical factors and inherited defects of connective tissues are the most frequent causes of this disease. Fibroblasts derived from women presenting striae distensae lesions were included into collagen gels to study their mechanical behavior: capacity to contract free floating lattices and to produce isometric force in tense lattices. To measure the retracted lattice diameter, the culture dishes were placed on a transparent metric scale. An isometric force system was used to study quantitatively the forces developed during lattice contraction. alpha 2 beta 1 integrins expression (transmembrane receptors) was evaluated by flux cytometry. Striae distensae fibroblasts contract collagen gels slower than normal human fibroblasts but the final contraction is similar. They produce a greater isometric force which is associated with enhanced alpha 2 beta 1 integrins expression. By their mechanical properties, striae distensae fibroblasts appear as a different population from normal fibroblasts. PMID- 11938561 TI - [Diminution of the in vitro and in vivo alloreactive response by association of mature dendritic cells and anti-CD4 monoclonal antibodies in the rat]. AB - Using a fully mismatched model recipient Lewis-donor Wistar-Furth rats, we showed that the association donor mature (spleen) dendritic cells with a non depleting anti-CD4 monoclonal antibody can induce a more pronounced decrease of alloreactivity in mixed splenocyte reaction than anti-CD4 alone. These results are observed in vitro and 30 days after injection to the Lewis rats. Such data suggest that anti-CD4 monoclonal antibody can guide mature dendritic cells, usually involved in acute rejection, towards tolerance. The mechanisms are still to be resolved. PMID- 11938563 TI - [Vibration after effect during human walking]. AB - Muscular vibration effects could usually persist after the end of the stimulation. The extinction of this after-effect was studied during human walk. Six subjects performed a locomotor task (six successive steps) under control, and after bilateral Achilles' tendon vibration (70 Hz). The step frequency was enhanced (diminution of the stance phase duration) whereas the stride length tended to decrease. The relative duration of the stance phase reached back normal values with repetitions. These results demonstrated that vibration-induced effects persevere after the end of the stimulation. In addition, owing to the dynamic of the re-adaptation process, they suggested that the muscular proprioceptive input could participate in both maintaining and regulating the locomotor rhythm. PMID- 11938562 TI - [Multimeric receptors, new targets for the pharmaceutical industry]. AB - Cytokines and growth factors are reported to play a pivotal role in many pathologic conditions. Some recombinant proteins have demonstrated powerful therapeutic activities for example in arthritis for anti-TNF (antibody and soluble receptor) or in cancer for interleukin2 and are now available for clinical use. However limitation in production, treatment condition for use and final high costs have encouraged pharmaceutical industry to develop research of small synthetic compounds which can replace or inhibit natural ligands. During the last 5 years a series of publication have demonstrated that small peptides (up to 20 amino-acids) and non-peptide synthetic compounds may replace in vitro as well as in vivo some cytokine and cell growth factors. Selection of these compounds have used new technique of screening including phage display and gene reporter assays. Currently selected compounds mainly act as agonist and stimulate transduction signals in the target cell as do natural ligands. These results have demonstrated that at least for some cytokine and cell growth factor protein biological action may be mimicked by small molecular weight synthetic compound. Next steps will deal with selection of drug candidates able to be studied at the clinical level. PMID- 11938564 TI - [BSE and scrapie diagnosis in Germany]. AB - The detection of pathological prion protein is considered as pathognomonic for the diagnosis of transmissible spongiform encephalopathies. According to the EU regulations cattle older than 30 months of age (Germany 24 months) and slaughtered for human consumption must be tested by using BSE rapid tests. Likewise must be fallen stock and clinically affected animals. This article gives an overview over the diagnostic hierarchy and organization of the diagnostic system for BSE and scrapie in Germany. All suspect cases found by rapid testing are reinvestigated and clarified by the National reference laboratory for these diseases which is part of the recently founded Institute of Novel and Emerging Infectious Diseases at the Federal Research Centre for Virus Diseases of Animals located on the isle of Riems. Until the end of 2001 130 BSE cases were confirmed out of 230 submissions. PMID- 11938566 TI - Studies on contamination of beef with tissues of the central nervous system (CNS) as pertaining to slaughtering technology and human BSE-exposure risk. AB - Contamination of beef by tissues of the central nervous system (CNS) due to slaughter technology causes some concern considering the potential health hazard by food borne exposure to the infectious agent of BSE. The present study was designed to quantify the extent of CNS contamination as pertaining to stunning and splitting technology. Of the 726 animals 48 contained a total of 58 emboli like particles in lungs and/or right ventricles. The incidence of emboli-like particles was found to be slightly higher in animals slaughtered without pithing (5.9%) than in the animals slaughtered with pithing (4.1%). Of the 58 emboli-like particles only two were positive in the anti-NSE western immunoblotting (0.3% of the 726 animals). The immuno reaction of these NSE-positive particles was several orders of magnitude lower as obtained by pure brain material. The microscopical analysis of the two NSE-positive emboli-like particles for presence of CNS-like tissues was negative. Following splitting of carcasses by sawing with and without prior removing the spinal cord we found NSE-positive reactions in 32% and 17% of the samples, respectively. The immuno reaction, however, was predominantly comparable to standard material containing less than 0.5% CNS. Overall the results show that CNS contamination of bovine carcasses cannot be excluded by current slaughter technology. However, the additional human BSE-exposure risk can be judged to be at least minor when considering extent of contamination, dilution effects and BSE-testing. PMID- 11938565 TI - Detection of CNS and PrPSc in meat products. AB - Several methods for the detection of tissues of the central nervous system (CNS) in meat products have been developed and partly validated for use in official food control as pertaining to human BSE-exposure risk. So far, however, methods for the detection of abnormal prion protein (PrPSc) were not evaluated for their potential applicability to the matrix of heat treated meat products. We developed a micro technological procedure for the preparation of meat products suitable for high security laboratories as masses were 6 to 8 orders of magnitude lower than in conventional meat technology. Thus it was possible to produce standard micro sausages containing defined amounts of bovine BSE-positive brain. This material showed all characteristics of normal meat products and a homogeneous distribution of brain as indicated by NSE and GFAP western immunoblotting and GFAP immunometric analyses. Using a commercially available and certified immunometric assay for detection of PrPSc in untreated brain it was possible to detect BSE positive CNS down to a content of 0.25% in heat treated meat products. We found a high correlation between PrPSc OD-values and CNS content and linearity up to 10% CNS. In 30 samples of retail meat products no sample transgressed the official cut off value for untreated bovine brain. Further studies are needed to show whether an increase of sensitivity in PrPSc detection from the meat product matrix is possible, in particular by optimisation of the extraction procedure. PMID- 11938567 TI - [Cellular nucleic acids in serum and plasma as new diagnostic tools]. AB - Currently, the diagnosis of bovine spongiform encephalopathy is only possible in the brain stem of dead animals. Protease resistant prions are detected in the obex region of the brain stem. However, from a veterinary medical and agricultural point of view the development of an in vivo detection assay is of utmost importance. Because infectious prions are detectable relatively late in the central nervous system during an infection, efforts are made searching for surrogate markers. Besides neuronal proteins that are released into the liquor and blood during neurodegenerative processes or other neuronal diseases, cellular nucleic acids circulating in the plasma or serum are an absolutely new approach for the detection of infectious diseases. PMID- 11938568 TI - [Measures preventing BSE-contamination during the slaughter of cattle]. AB - SRM-regulations and the prohibition of pithing have removed major risks of spreading BSE-infection. Traditional slaughter technology, especially captive bolt stunning, head handling and carcass splitting nevertheless still provide non negligible risks for contamination with the BSE-agent if present in cattle, and should therefore be replaced by safer techniques. However, alternative methods like electrical stunning or removal of the spinal cord prior to splitting the carcass cannot yet be considered a reliable and practical option. Surface contamination could be prevented altogether by abandoning the practice of carcass splitting and by removing the vertebral column while still connected to the head, although this would result in disadvantages for post mortem inspection. PMID- 11938569 TI - Detection of tissues of the central nervous system (CNS) as specified risk material (SRM) in meat products by means of gas chromatography-mass spectrometry (GC-MS). AB - Determination of specified risk material (SRM) in processed meat products was performed by quantification of brain specific fatty acids using gas chromatography-mass spectrometry (GC-MS). Results from SMP (internal standardised meat products) based analyses showed that absolute concentrations of CNS are correlated (r2 = > 0.97) with the contents of the CNS typical fatty acids docosahexaenoic acid (C 22:6), nervonic acid (C 24:1), lignoceric acid (C 24) and cerebronic acid (C 24oh). GC-MS detection limits were measured at 0.01% CNS. The cut off value was calculated at 0.39% (w/w) CNS in SMP. In a controlled blindfold experiment we were able to identify correctly all positive and negative SMP samples, respectively. Our results indicate that GC-MS based SRM detection may serve as a reference method for immunochemical and immunohistochemical determination of SRM in processed meat products. PMID- 11938570 TI - [Protective measures taken with regard to BSE under veterinary law]. AB - BSE was established for the first time in 1986 as a separate disease complex. Since 1989 measures to protect human and animal health have been adopted at Community level and under German law. The article describes the most important provisions governing the prevention, control and eradication of TSE. It addresses in detail the ban on feeding, active monitoring of BSE, active monitoring of small ruminants, measures taken after the detection of BSE, the removal and destruction of specified risk material and briefly addresses trade bans and restrictions. PMID- 11938571 TI - [Transmissible spongiform encephalopathies (TSE): old diseases with new explosive force]. AB - In view of the first 64 BSE cases (date: 11.5.01) in German cattle herds an overview on TSE and their similarities and differences regarding clinic, pathogenesis and pathology is given. The mechanism of the unconventional agent, an infectious protein (prion), is explained based on the prion model of Stanley Prusiner. The knowledge on transmission, incubation time, host specificity as well as resistance and immunity drawn from experimentally infected animals is discussed. Thus, after oral infection prions are transported by lymphocytes from the stomach-intestinal tract to the spleen. The way to the CNS is still unknown. The presumption for crossing the species barrier is twofold: first the prions of different species have to be biochemically homologous and a genetical disposition has to exist. This is the case for BSE and the new variant of Creutzfeldt-Jakob Disease (vCJD). There is evidence that in Great Britain so far 97 (date: 30.3.01) young people acquired vCJD due to consumption of food that contained bovine risk material. Regarding the infectious prion dosis brain, spinal cord and lymphoid tissues are regarded to be most dangerous. The principle of the BSE-test, its evidence as well as steps for prevention and control of BSE are presented. PMID- 11938572 TI - [The prion hypothesis and the human prion diseases]. AB - Our understanding of the pathogenesis of the transmissible spongiform encephalopathies (TSE) has made terrific headway over the past 40 years and some scientists are even of the opinion that this group of diseases belongs to the neurodegenerative syndromes best understood. On the other hand, the investigation of TSE has led to a multitude of unexpected and surprising results and consequently has initiated impassioned discussions among scientists. Although the human forms of TSE are very rare, the wildfire-like spread of the bovine spongiform encephalopathy (BSE) raises the pressing question as to whether BSE is communicable to humans. This overview summarizes some current hypotheses about the nature of the infectious agent and about the pathogenesis of the damage of the central nervous system. PMID- 11938573 TI - [The clinical diagnosis of BSE]. AB - Diagnosis of Bovine spongiform encephalopathy (BSE) is confirmed by specified laboratory methods on brain material. On the other hand clinical signs of manifest BSE are quite obvious. The first part of this paper describes case histories, clinical signs, laboratory findings and the most common differential diagnoses. On the basis of the data of actual prevalence in Germany, the role of clinical examination in eradication of BSE is dealt in the second part. Clinical diagnosis is a very sensitive and specific method when there is a high prevalence. According to the data from December 2000 to November 2001 prevalence in Germany was beyond 1 BSE case per 100,000 cattle or 3 cases per 100,000 cows. This very low prevalence decreases rapidly sensitivity and specificity of the diagnosis made by clinical examination. Therefore the main focus of field diagnostics has to be laid on specified laboratory diagnostic methods. On the other hand prevalence of BSE-positive cattle is distinctly higher in the group of animals slaughtered in cases of illness or emergency than in cattle slaughtered on the regulatory bases. Nevertheless every veterinary practitioner should be aware of the clinical picture of BSE, clinical examination-routine and differential diagnosis, because occurrence of BSE is still possible in any dairy herd. At the moment it is not possible to make any statement if eradication of BSE can be reached in future. PMID- 11938574 TI - [An inherited hemostatic disorder as the cause of menorrhagia]. AB - Two women aged 39 and 30 years were investigated for possible coagulation disorders because of menorrhagia, anaemia and postoperative haemorrhages. These investigations revealed that they had type 1 Von Willebrand's disease. During the treatment with desmopressin, factor VIII and Von Willebrand factor (VWF) activity normalised. About one third of the patients referred to a gynaecologist for menorrhagia have an inherited bleeding disorder, of which type 1 Von Willebrand's disease is the most prevalent. Once a gynaecological cause of the menorrhagia has been excluded, or in the case of an increased risk on the basis of the medical history, a limited haemostatic investigation can establish or exclude an inherited coagulation disorder. For women with a coagulation disorder the menorrhagia can be treated. Surgical interventions can be carried out safely following treatment with desmopressin or factor VIII/VWF concentrates. PMID- 11938575 TI - [The advances of HIV in the Netherlands: indications but still no definite answers]. AB - Investigations in the province of Limburg demonstrate an increasing HIV prevalence and a high prevalence of injecting and sexual risk behaviour as well as a substantial risk of HIV transmission to the general population. It is recommended that HIV surveillance be directed towards 'bridge populations' such as prostitutes, their customers and immigrants from HIV endemic countries, instead of just drug users. In view of the apparent increase in sexual risk behaviour and sexually transmitted diseases in homosexual men (possibly due to a reduced fear of the disease because of the availability of anti-HIV medication) surveillance should be intensified in that area as well. Only then will it be possible to ascertain whether or not the incidence of HIV in the Netherlands is increasing. PMID- 11938576 TI - [Class substitution of medicines--between cost reduction and the position of professionals in health care]. AB - Class substitution (with cost reduction as the purpose) is only possible if there are equivalent drugs in terms of the pharmacological mode of action and safety both at the individual and population level, as well as per indication. Whether two drugs are found to be equivalent, is in part the consequence of the chosen clinically relevant margin in which equivalency can reasonably be assumed and the accuracy with which the outcome measure could be measured. Good data for carrying out class substitution will in many cases be lacking. It is for the health professional to decide which preparation is indicated for a certain patient whereby the pharmacist provides pharmacotherapeutic information and checks the prescribed treatment. The provision of education in medical and pharmaceutical curricula is the most effective manner for limiting drug costs. PMID- 11938577 TI - [Class substitution as an instrument of efficiency in the supply of medicines: possibilities and limitations]. AB - Class substitution is the substitution of a drug by another cheaper drug from the same pharmacological drug class to save costs. This principle is applied in drug formularies and reference-based drug-pricing systems. Class substitution should not only consider similarities in pharmacological mechanisms of action but should also assess whether the different drugs within the class are equivalent at a population level with respect to efficacy, effectiveness, applicability, and safety. The implementation of class substitution requires protocols as well as a monitoring system to evaluate compliance with the protocols and their effects on drug costs. In addition, specific studies are needed to establish whether undesirable effects occur at the patient level. PMID- 11938578 TI - [Non-scrotal testes; first line of management]. AB - For non-scrotal testes a distinction can be made between retractile testes (completely descended and normally developed but sometimes situated subcutaneously in the groin area), retained testes (testes cannot be brought into the scrotum or this can only be achieved using light manual pressure) and ectopic testes (lying outside of the descent trajectory). It is estimated that 0.7-0.8% of all boys have as yet undescended testes. The first few days after the birth are the most suitable for testing and registration, as then the cremaster reflex is absent. Registration should take place in both the youth healthcare file and in the 'growth book' for the parents. Retractile testes do not require treatment. There is no consensus concerning the treatment of (possible) acquired nonscrotal testes. For undescended testes the management depends on previous testes localisations. For ectopic testes and testes that have never been scrotal, a referral for surgical treatment should be made prior to the second birthday. Orchidopexy (a better description is orchidofuniculolysis followed by orchidopexy) is only justified in the case of testes which have never descended. In the case of a clear indication, the general practitioner should make a prompt referral (before the second birthday) and in other cases assurance should be provided and an expectant policy adopted until puberty. PMID- 11938579 TI - [Diagnostic images (81). A man suffering from intermittent stomach pain and weight loss]. AB - A 63-year-old male was referred because of colicky pain due to an intussusception of a Meckel's diverticulum and of the ileum into the cecum. PMID- 11938580 TI - [Previous testicular position in boys who underwent orchidopexy due to undescended testis at the Alkmaar Medical Center, the Netherlands, during the period 1986-1999]. AB - OBJECTIVE: To determine the previous testicular position in boys, in whom orchidopexy was performed for undescended testis. DESIGN: Retrospective, descriptive. METHOD: All boys, aged 0-18 years, who underwent orchidopexy in the Alkmaar Medical Centre, the Netherlands, during the period 1986-1999, were studied. The following information was obtained from the hospital medical records: indication for operation, date of the operation, laterality (unilateral or bilateral), the surgical findings and whether previous testicular position played a role in the decision to perform orchidopexy. For each boy who underwent orchidopexy for undescended testis, previous testicular positions up until the date of the operation were obtained from the appropriate youth health care institutions. RESULTS: Hospital records were available for 851 boys who had undergone orchidopexy. The operation for undescended testis was performed in 717 boys and previous testicular positions were obtained for 565 boys. On a per testicle basis, 707 operations were carried out (142 bilaterally, 205 left-sided, 218 right-sided). From these 707 testes, a previous intrascrotal position was found at least once in 572 (80.9%), at least twice in 493 (69.7%) and at least three times in 419 (59.3%); 135 (19.1%) testes had never been intrascrotal. The majority of previously undescended testes were operated on at 3 years of age; most operations on previously descended testes were performed at 10.5 years of age. For 344 (48.7%) out of 707 testes, previous testis localisation was known in the hospital's medical records, for 96 (13.6%) testes registration was unclear and in 267 (37.8%) testes it was not reported. In 8 (1.4%) boys, testis registration after the birth was used on referral to document previous testicular position. CONCLUSION: In total 80.9% of all orchidopexy operations were performed on testes that had previously been diagnosed as having descended normally. These probably included retractile testes as well as acquired forms. In 51.3% of the cases, previous testicular position was not known in the hospital's medical records at the time of operation. PMID- 11938581 TI - [HIV prevalence among injecting drug users in South Limburg, 1994-1998/1999: increasing trend in Heerlen, not in Maastricht]. AB - OBJECTIVE: To determine the prevalence of HIV infection and gain insights into the risk behaviour among injecting drug users (IDU) in Heerlen and Maastricht, the Netherlands. DESIGN: Descriptive inventory. METHOD: In 1994, 1996 and 1998/1999 a questionnaire on risk behaviour was obtained from 866 IDUs in Heerlen and Maastricht, and 858 saliva and serum samples were also obtained for testing on HIV antibodies. RESULTS: In the total region of Heerlen and Maastricht, no significant change in HIV prevalence was found (1994: 10%, 1996: 12%, 1998/1999: 14%). However, in Heerlen the HIV prevalence was demonstrated to have increased significantly (1994: 11%, 1996: 16%, 1998/1999: 22%), whereas in Maastricht HIV prevalence was constant (1998/1999: 5%). The percentage of participants that injected drugs in the six months preceding the survey decreased from 80% in 1994 to 63% in 1998/1999. Injection-related risk behaviour slightly decreased: in 1998/1999 14% of the participants had borrowed used syringes or needles in the last 6 months. Additionally, inconsistent condom use with steady and casual sexual partners was reported. Of the participants, 86-89% did not always use condoms with steady partners and 49% with casual partners. DISCUSSION: In view of the increasing HIV prevalence among IDUs in Heerlen, the persistent injecting and sexual risk behaviour and the large number of IDUs who have sexual contact with non-users in Heerlen and Maastricht, there is a substantial risk of HIV transmission both within the IDU community and to the general population. PMID- 11938582 TI - [Psychosis during and after disulfiram use]. AB - A 47-year-old man was registered for treatment of his alcohol abuse problem. Disulfiram was administered as part of the treatment, the intake of which was supervised by his wife. During the treatment with disulfiram and two weeks after the withdrawal of this treatment he developed a psychosis. He stated that he had not taken alcohol. During the second psychotic period he was successfully treated with antipsychotic drugs. Afterwards it was discovered that his family history was positive for schizophrenia; it is therefore possible that the patient was more vulnerable for a psychosis due to disulfiram use. Although a psychotic reaction during and after disulfiram therapy is not common, it should nonetheless be taken into consideration when prescribing the drug. PMID- 11938583 TI - [Hallucinations caused by paroxetine taken together with a levodopa-carbidopa preparation]. AB - A 79-year-old woman suffering from Parkinson's disease, for which she was taking a levodopa-carbidopa preparation, was prescribed the selective serotonin reuptake inhibitor paroxetine 20 mg once daily. After taking the first tablet she started to suffer from visual hallucinations. Once she stopped taking the paroxetine, the hallucinations ceased immediately. A link between the paroxetine and the hallucinations seemed likely, with a possible interaction between the paroxetine and the levodopa-carbidopa combination. PMID- 11938584 TI - [Karl Landsteiner (1868-1943) and the specificity of serological reactions, one hundred years ago and now]. AB - One hundred years after his discovery of the AB0 blood groups, the monumental work of Austrian physician Karl Landsteiner (1868-1943, Nobel Prize 1930) on the specificity of serological reactions is widely recognised. His work on isoantigen reactions included the discovery, together with Levine and Wiener, of the Rhesus factor. Landsteiner did much to develop our understanding of autoantibody reactions and also contributed insights into the aetiology of infectious diseases that were rampant in his time. His work in immunochemistry included the discovery of hapten antigenicity. However, most of all he wanted to be remembered for his theory on the specificity of serological reactions, which he had strongly substantiated with his own work. Landsteiner suffered from the shortages in Vienna at the end of World War I. He and his family moved to the Netherlands in 1919. There he worked as pathologist at the Red Cross Hospital in The Hague until his move to the Rockefeller Institute in New York in 1923. Nowadays, his work is celebrated for its numerous excellent contributions to the fields of transfusion and transplantation medicine, population genetics, immunology, and infectiology. PMID- 11938585 TI - [Developments in the area of hearing aids]. PMID- 11938586 TI - [Arguments for providing serum screening for Down syndrome for all pregnant women]. PMID- 11938587 TI - BS-SEM evaluation of the tissular interactions between cortical bone and calcium phosphate covered titanium implants. AB - The improvement of the reliability of the contact between the osseous tissues and the implant materials has been tested by recovering the metallic implants with ceramic materials, usually calcium phosphates. In our study, the calcium phosphate recovering layers were deposited by means of a pulsed-laser deposition technique. Our aim was to to evaluate the tissue interactions established between cortical bone and titanium implants covered by five different layers, ranging from amorphous calcium phosphate to crystalline hydroxyapatite, obtained by altering the parameters of the laser ablation process. The surgical protocol of the study consisted in the simultaneous implantation of the five types of implants in both the tibial dyaphisis of three Beagle dogs, sacrificed respectively one, two and three months after the last surgical procedures. After the sacrifice, the samples were submitted to a scheduled procedure of embedding in plastic polymers without prior decalcification, in order to perform the ultrastructural studies: scanning microscopy with secondary and backscattered electrons (BS-SEM). Our observations show that both in terms of the calcified tissues appearing as a response to the presence of the different coatings and of time of recovering, the implants coated with crystalline calcium phosphate layers by laser ablation present a better result than the amorphous-calcium-phosphate coated implants. Moreover, the constant presence of chondroid tissue, related with the mechanical induction by forces applied on the recovering area, strongly suggests that the mechanisms implied in osteointegration are related to endomembranous, rather than endochondral ossification processes. PMID- 11938588 TI - Comparative biomorphologic analysis about three dentinal adhesives of last generations. AB - The aim of this work consists in a comparative biomorphological analysis of the properties of infiltration and of adhesion to dental tissues of three among the more used enamel dentinal adhesives of the last generation known with the commercial name of Syntac, Excite and Prompt. The results have given evidence that Syntac has got short adhesion, Excite has got good capacity of infiltration and moderate adhesion, Prompt seems to possess a capacity of infiltration equal to Excite's one, but a better adhesion besides an easier modality of use. PMID- 11938589 TI - Klammt's Elastic Open Activator: Ricketts' cephalometrics results. AB - The purpose of this study is to investigate the orthodontic and orthopaedic real effects of the Klammt's Elastic Open Activator (EOA) in 25 Class II Division 1 patients in growing period. We wanted to determine statistically the cephalometrics changes produced in the patients, comparing the lateral cranium teleradiographies we took for the diagnosis with the ones we took at the end of treatment. At the end of this study we confirm that by using the EOA we obtained the desired effects, especially reducing the molar relation 2.53 mm and the overjet 2.56 mm. The EOA corrected the inclination and protrusion of incisors, although we cannot avoid the use of fixed appliances to round off. The reduction of 2.48 mm of facial convexity stands out as the most important skeletal effect; the facial depth angle increases 0.8 degree, and the maxillary depth decreases 1.16 degrees. The length of the mandibular corpus also increases 6.7 mm, although this change is mainly due to the growth of the patient. The changes in the aesthetic profile do not stand out. PMID- 11938590 TI - Effects of Nd: YAG laser irradiation on the root canal wall dentin of human teeth: a SEM study. AB - The purpose of this study was to observe the morphological and histological changes on the root canal walls after Nd:YAG laser application. Twenty vital, recently extracted single-rooted human teeth were used for this study. Root canals were cleaned and shaped by a conventional step-back technique--by means of k files up to a 20 k-file type at working length--and subsequently shaped by Ni Ti root-canal rotary instrumentation up to 30/06 and irrigated with 2.5% hypochlorite solution. Ten teeth (control group) were left unlased, while the other ten teeth were irradiated with Nd:YAG laser by means of a 320 microns fibre inserted in the root canal at 1 mm from the apex with a power of 1.5 Watt and a frequency of 15 pps for five seconds in retraction with rotating movements. The control specimen showed debris and smear layer on the root canal surface obscuring the dentin tubules. The root canal walls irradiated with Nd:YAG laser showed a clear glazed surface, some open dentinal tubules and some surface craters with cracks. Such results confirm that smear layer and debris are removable with Nd:YAG laser, however clearing all root canal walls is still difficult and, if the energy level and duration of application are inadequate, a certain degree of thermal damage and morphological changes in dentin structure are observable. PMID- 11938591 TI - [The first appearance of Meckel's cartilage in the fetus]. AB - Meckel's cartilage plays an important role in the topographical organisation and in the differentiation of the facial structure during the embryonal and even much later during the foetal period. Our observations on serial sections carried out in two human foetuses aged 12 and 16 weeks indicate that the two dorsal (tympanic) and ventral (mandibular) branches of Meckel's cartilage are perfectly defined at 16 weeks. In the dorsal branch, the primordia of the incus and of head of the malleus are still composed on non-ossified cartilage. In the ventral branch, it is also possible to describe at 16 weeks three posterior, medial and anterior parts which are composed of cartilage. The initiating role played by the ventral part of Meckel's cartilage on the ossification of the mandible leads during the embryonal period to the formation of the mandibular primary growth center, which is therefore clearly defined in our first stage at 12 weeks. The partial fibrous evolution and the regression of the major part of the ventral branch of Meckel's cartilage only start after 16 weeks of intrauterine life. PMID- 11938592 TI - Clinical and electromyographic examinations of patients with midbrain and cerebellar tremor. AB - The aim of this study was to re-evaluate the controversial clinical and electromyographic findings of cerebellar and midbrain tremors. Neurological examination and surface electromyographic tremor recordings were performed on 46 patients (24 with midbrain and 22 with cerebellar tremor). Our results revealed that the midbrain tremor has involved the hand, sometimes the ipsilateral leg and regions outside the limbs. Usually it was combined rest, postural, kinetic and intention tremor, the amplitude of each next tremor progressively increased. The tremor had high amplitude, low frequency and typical long burst duration. The cerebellar tremor was quite different, as it was predominantly postural and kinetic and involved the upper limbs. The tremor had synchronous electromyographic pattern, high frequency, low amplitude and short burst duration. In conclusion although the cerebellar and midbrain tremors are caused by involvement of very close anatomical regions and sometimes one type of tremor may become into another, they have quite different clinical and electromyographic presentations. PMID- 11938593 TI - Reduced sensory nerve conduction velocity of the distal part of the radial nerve among patients with vibration syndrome. AB - The present study was aimed at clarifying the effects of vibration syndrome (VS) on the distal part of the radial nerve, which is rarely affected by chronic entrapment. Thirty-one patients with VS due to previous exposure to vibration and 18 age-matched controls were examined for sensory nerve conduction velocities in the distal part of the radial nerve (SCV-DR) in the dorsal side of the hand. The patient group was divided into two subgroups, one of those with vibration-induced white finger (VWF) in the thumb or index finger where sensory nerves are innervated by the radial nerve (N = 21) and the other without VWF (N = 10). SCV DR among the patients was significantly slower than in the controls (t = 3.55, p = 0.00138). The ANOVA of SCV-DR for the three groups showed F2, 46 = 5.25 (p = 0.0088). A significant difference was found between the controls and the VWF (+) group (p = 0.0233), but not between the controls and the VWF (-) group (p = 0.478) by multiple comparison using Scheffe's method. These findings suggest that VS affects the radial nerve with a direct vibration effect and/or a co-operative effect with direct circulatory disturbance manifested by VWF. PMID- 11938594 TI - Surface compound action potentials recorded from different locations on the anterior tibial muscle. AB - Supramaximal CMAPs to peroneal nerve stimulation at the knee were recorded from 5 locations on the anterior tibial muscle in 24 patients (30 muscles). The active recording locations were: midpoint of the muscle belly, at 4 cm distal and proximal to it, and at 2 cm lateral and medial to it. Reference electrode was at the medial malleolus. CMAP duration, amplitude, and area were measured, and ratios of their corresponding minimum to maximum values were computed. Thirteen patients (15 muscles) had normal nerve conduction and needle EMG studies. Mean ratios of minimum to maximum values were duration = 0.89, amplitude = 0.67, and area = 0.75. Eleven patients (15 muscles) had abnormal studies, and the mean ratios of minimum to maximum values were duration = 0.87, amplitude = 0.66, and area = 0.71. CMAP duration, unlike amplitude and area, appears least likely to be influenced by the recording electrode location, and is a more stable and reproducible measure during nerve conduction studies. The maximum (or minimum) values in the CMAP duration, amplitude, and area frequently do not coincide to one electrode recording location. A majority of the CMAPs recorded from different muscle locations had an initial negative phase, suggesting that the endplate zones of the anterior tibial muscle are dispersed rather than concentrated in a small region. PMID- 11938595 TI - Comparison of the antidromic sensory nerve action potentials obtained from all fingers on the same hand. AB - Supramaximal compound sensory nerve action potentials (CSNAPs) were recorded antidromically from five fingers of the same hand to electrical stimulation of the median, ulnar, and radial nerves in 17 normal subjects. The mean amplitudes of the median CSNAPs from the thumb, index, and middle fingers were similar in values. There was minimal radial sensory contribution to the thumb, with the mean radial CSNAP amplitude about one-fourth the size of the median CSNAPs. The mean median and ulnar CSNAP amplitudes from the ring finger were approximately half, and the mean ulnar CSNAP amplitude from the little finger was approximately 85%, of the median CSNAP amplitudes from the other fingers. The median, ulnar, and radial sensory nerve conduction velocities were close in values. There appears to be less variability in the population normative data when the size of the CSNAPs is expressed in terms of amplitude ratio rather than in absolute amplitude value. PMID- 11938596 TI - Characteristic appearances of the H-reflex and F-wave with increased stimulus intensity in patients with cerebrovascular disease. AB - We experienced H-reflex may be evoked with supramaximal stimulation in patients with cerebrovascular disease (CVD). In this study, we investigated the relationship between the characteristic appearances of H-reflex and F-wave with increased stimulus intensity and neurological signs. We examined the H-reflex and F-wave of the affected arm with increased stimulus intensity during muscle relaxation in 31 patients (17 males and 14 females) with hemiparesis caused by CVD. Mean patient age was 56.0 (range 30-82) years. 30 healthy subjects, mean age of 56.2 (range 28-80) were investigated using the same method as controlled group. H-reflex and F-wave with increased stimulus intensity after the median nerve stimulation at the wrist were recorded from the opponence pollicis muscle on the affected side in patients with CVD or right arm in the healthy subjects. Appearance patterns of the H-reflex and F-wave with increased stimulus intensity was separated into four types. Type 1: F-wave appeared with increased stimulus intensity, but there was no H-reflex. Type 2: H-reflex and F-wave both appeared with increased stimulus intensity, but the F-wave followed disappearance of the H reflex with increased stimulus intensity. Type 3: H-reflex and F-wave both appeared with increased stimulus intensity, but the F-wave appeared during the H reflex with increased stimulus intensity. Type 4: Only the H-reflex appeared with increased stimulus intensity, but there was no F-wave. Neurological findings including muscle tone and tendon reflex were also evaluated. Findings on muscle tone and tendon reflex were classified into increased (markedly, moderately and slightly), normal and decreased. Results were analyzed as follows; 1) The characteristic appearances of H-reflex and F-wave in the healthy subjects and 2) The relationship between characteristic appearances of waves with increased stimulus intensity and neurological signs in patients with CVD. 1) Pattern of the H-reflex and F-wave with increased stimulus intensity in all healthy subjects were type 1. 2) Patterns of the H-reflex and F-wave with increased stimulus intensity in patients with markedly increased muscle tone and tendon reflex were almost always type 4. Those with moderately increased signs in this parameter demonstrated type 2 or 3. Those with slightly increased signs in this parameter demonstrated type 1 or 2. Those with normal or decreased signs were type 1. These findings suggest that the characteristic appearances of the H-reflex and F-wave was influenced by grade of neurological signs. PMID- 11938597 TI - Electromyographic study of the masseter muscle in individuals with Class II malocclusion. AB - The objective of this work was to study the action of the masseter muscle through electromyography and to analyze the difference of action potential generated by several pre-established jaw movements in individuals with Class II malocclusion. It was studied nine young individuals from both sexes, with ages ranging from 17 to 35, with no history of orthodontic treatment and/or having some kind of alteration in the temporomandibular joint (T.M.J.). In order to collect the electromyographic signs we have used a TECA TE-4 Electromygraph and surface electrodes type Beckman. The data were submitted to nonparametric statistical analysis by Friedman test. The response variables left occlusive contact, free protraction, protraction with occlusive contact, incisor mastication, right molar mastication and forced central occlusion movements showed significant outcomes. Thus we have concluded that as the occlusive balance suffers alterations, the malocclusions become predominant and the masseter muscle undergoes functional and structural modifications. PMID- 11938598 TI - Electromyographic validation of the trapezius and serratus anterior muscles in the rowing and frontal-lateral cross, dumbbells exercises. AB - The trapezius and serratus anterior muscles were studied in four modalities of rowing exercises executed with two grips, middle and closed, in comparison with the four different modalities of frontal-lateral cross, dumbbells exercise. It was used 24 male volunteers, 18 to 25 years old using a 2-channel TECA TE 4 electromyograph and Hewlett Packard surface electrodes. The results showed that TS acted in a higher significant way in all the modalities of rowing than in the supine lateral raise, inclined supine lateral raise and reverse supine lateral raise, dumbbells exercices, and demonstrated no standing frontal-lateral cross, dumbbells. The SI acted more significantly difference among all the execution modalities of rowing and the inclined supine lateral raise, dumbbells exercises than in all the rowing exercises; even though the activity levels do not make us suggest them as an excellent group of exercises for the development of this muscle. PMID- 11938599 TI - Stroke recovery induced by coordination dynamic therapy and quantified by the coordination dynamic recording method. AB - Based on measurements of relative phase and frequency coordination of time-space distributed firing of single neurons and neuron assemblies in the human nervous system, the concept of coordination dynamics to describe the integrative functions of the human central nervous system (CNS) and the finding of neurogenesis in the human CNS, the coordination dynamic therapy has been developed with which it is possible to repair the lesioned or malfunctioning human CNS. With the developed coordination dynamic recording method the organization (the coordination dynamics) of the CNS can be measured on-line non invasively. By measuring the present coordination dynamics at different times with ongoing coordination dynamic therapy when exercising on a special coordination dynamic therapy device, the progress in re-organization and repair of the lesioned CNS can be quantified. In this paper it is reported that in 8 patients following stroke the lesioned CNS could partly be repaired by coordination dynamic therapy. The repair was quantified by the improvement of walking, hand function and the direct measurement of CNS organization (the coordination dynamics). In 4 patients the improvement of CNS organization (coordinations dynamics) was 70 +/- 14% within 3 months of therapy. In a patient where the intensive coordination dynamic therapy (at least 4 hours therapy with more than 15,000 coordinated movements per day) lasted longer than one year, also the difficult repairable hand functions could substantially be improved. It is shown that in the healthy CNS of the author the coordination dynamics improved in the short-term memory by 20% within 10 min and in a stroke patient by 70% within 30 min, when exercising on the special coordination dynamic therapy device. As in other learning therapies (school) it is believed, that by repeated learning of movements, vegetative and higher mental functions in the short-term memory, the learned CNS functions will also go into the long-term memory, which means the CNS has been reorganized. Improvements in the short-term memory are used to motivate the patient to perform a therapy. PMID- 11938600 TI - Consumer health care finances and education: matters of values. PMID- 11938601 TI - Testing your diagnostic skills (59). Case 1. Pyogenic granuloma. PMID- 11938602 TI - Testing your diagnostic skills (59). Case 2. Central giant cell granuloma. PMID- 11938603 TI - Nuclear terrorism to cause ultimate health crisis. PMID- 11938604 TI - Functional consequences of adult malnutrition in developing countries: a review. AB - Credible concepts of malnutrition depend critically on evidence that functional impairment differentiates variation in size from unacceptable abnormality. This paper reviews functional studies on capacity for work, mortality, reproductive outcomes, competitive success, and cultural preferences. Future research priorities should include the comparative significance of simple physical dimensions in distinct ethnic groups for long-term biological and socioeconomic outcomes, and the relationships between the biological and cultural significance of variation in physique. PMID- 11938605 TI - Physical activity and health-related physical fitness in Taiwanese adolescents. AB - The relationship between physical activity and health-related physical fitness was evaluated in 282 Taiwanese adolescents 12-14 years of age. The subjects were randomly selected from the 7th, 8th and 9th grades in two junior high schools in Taiwan. Physical activity was estimated as total daily energy expenditure and energy expenditure in moderate-to-vigorous physical activity from 24-hour activity records for three days, two week days and one weekend day. Health related fitness was assessed as the one-mile run (cardiorespiratory endurance), timed sit-ups (abdominal strength and endurance), sit-and-reach (lower back flexibility), and subcutaneous fatness (sum of the triceps, subscapular, suprailiac, and medial calf skinfolds). Physical activity is significantly and positively correlated with one-mile run performance and the sit-and-reach, but not with sit-ups and subcutaneous fatness. Overall, the strength of the relationships between estimated energy expenditure and specific fitness items in the total sample vary from low to moderate, with only 1% to 12% of the variance in fitness variables being explained by estimated energy expenditure. Comparisons of active versus inactive, and fit versus unfit adolescents provide additional insights. The more active (highest quartile) are also more fit in cardiorespiratory endurance and in the sit-and-reach than the less active (lowest quartile), and the more fit in the one-mile run (better time, lowest quartile) and the sit-and-reach (highest quartile) are more active than the less fit in each item, respectively. PMID- 11938606 TI - Body mass index, physical activity, dietary intake, serum lipids and blood pressure of middle-aged Japanese women living in a community in the Goto archipelago. AB - Mortality in the Goto archipelago region of the Nagasaki prefecture in Japan is higher than the Japanese average. In this study, we investigated dietary intake, habitual physical activities, systolic and diastolic blood pressure, serum total cholesterol and high-density lipoprotein cholesterol of middle-aged women in the Narao community in the Goto archipelago. We compared these parameters with a Japanese sample, and analyzed the results according to body mass index (BMI) and age. The mean BMI of Narao women was higher than that of the Japanese sample. Serum cholesterol and blood pressure of Narao women correlated with BMI. However, dietary intake and physical activities and not differ between normal- and over weight Narao women. The higher serum total cholesterol and diastolic blood pressure of Narao women, relative to the Japanese sample, could be explained by the presence of over-weight women in the Narao community. However, dietary and behavioral factors associated with higher BMIs could not be clarified in this cross-sectional study. PMID- 11938607 TI - Mechanomyogram and electromyogram responses of upper limb during sustained isometric fatigue with varying shoulder and elbow postures. AB - To investigate the behavior of mechanomyogram (MMG) and electromyogram (EMG) signals in the time and frequency domains during sustained isometric contraction, MMG and surface EMG were obtained simultaneously from four muscles: upper trapezius (TP), anterior deltoid (DL), biceps brachii (BB), and brachioradialis (BR) of 10 healthy male subjects. Experimental conditions consisted of 27 combinations of 9 postures [3 shoulder angles (SA): 0 degree, 30 degrees, 60 degrees and 3 elbow angles (EA): 120 degrees, 90 degrees, 60 degrees] and 3 contraction levels: 20%, 40%, and 60% of maximum voluntary contraction (MVC). Subjective evaluations of fatigue were also assessed using the Borg scale at intervals of 60, 30, and 10 sec at 20%, 40%, and 60% MVC tests, respectively. The mean power frequency (MPF) and root mean square (RMS) of both signals were calculated. The current study found clear and significant relationships among physiological and psychological parameters on the one hand and SA and EA on the other. EA's effect on MVC was found to be significant. SA had a highly significant effect on both endurance time and Borg scale. In all experimental conditions, significant correlations were found between the changes in MPF and RMS of EMG in BB with SA and EA (or muscle length). In all four muscles, MMG frequency content was two or three times lower than EMG frequency content. During sustained isometric contraction, the EMG signal showed the well-known shift to lower frequencies (a continuous decrease from onset to completion of the contraction). In contrast, the MMG spectra did not show any shift, although its form changed (generally remaining about constant). Throughout the contraction, increased RMS of EMG was found for all tests, whereas in the MMG signal, a significant progressive increase in RMS was observed only at 20% MVC in all four muscles. This supports the hypothesis that the RMS amplitude of the MMG signal produced during contraction is highly correlated with force production. Possible explanations for this behavioral difference between the MMG and EMG signals are discussed. PMID- 11938608 TI - Effect of postprandial posture on digestion and absorption of dietary carbohydrate. AB - The effect of postprandial body posture on digestion and absorption of dietary carbohydrate were examined through breath hydrogen test on 6 female subjects. During the experiment, the participants either sat on a chair or lay on their backs for the first 4 hr (from 08:00 to 12:00) after eating the test breakfast meal. They then remained sedentary on a sofa for 6 hr (12:00 to 18:00). Participants' end alveolar breath samples were collected for 10 hr (every 15 min from 08:00 to 12:30, and then every 30 min until 18:00). The experiment was conducted on two consecutive days using a randomized, crossover study design. The results demonstrated that in the supine position orocecal transit time of the test meal was significantly slower than in the sitting position (260 +/- 21 min and 238 +/- 20 min, respectively, p < 0.01). In addition, afternoon breath hydrogen excretion due to a partial malabsorption of dietary carbohydrate and its fermentation in the colon was significantly larger in the sitting position (144.0 +/- 24.1 ppm.hr) than in the supine position (110.0 +/- 26.1 ppm.hr, p < 0.05). These results support the hypothesis that there was a marked effect of postprandial body posture on the function of the digestive system. The present findings suggest that the postprandial supine position is preferable to the sitting position for the digestion and absorption of dietary carbohydrate. PMID- 11938609 TI - A cysteine protease inhibitor prevents suspension-induced declines in bone weight and strength in rats. AB - In this study, we examined the effects of a potent cysteine protease inhibitor, N (L-3-trans-carboxyoxirane-2-cabonyl)-L-leucine-4-aminobutylamide (E-64a), on bone weight and strength in tail-suspended rats. We first administered a vehicle or 4 or 8 mg/rat of E-64a to rats fed with a low calcium diet for 7 wks to determine effective doses of E-64a on bone resorption in vivo. Femoral cathepsin K-like activity and serum hydroxyproline level in rats fed with a low calcium diet were significantly higher than those in rats fed with a standard diet. The intraperitoneal injection of 8 mg/rat of E-64a to rats decreased their serum calcium and hydroxyproline concentrations after 3 to 6 hrs in parallel with changes in femoral cathepsin K-like activity, while 4 mg/rat of E-64a had weaker effects on these parameters. Based on these results, we injected 8 mg/rat of E 64a to tail-suspended rats twice a day for 2 wks and compared the results with those of treatment with 1 mg/rat of etidronate, a bisphosphonate, twice a week. In tail-suspended rats, femoral weight and strength, assessed by three-point bending test, significantly decreased from Day 5 to 21, while femoral cathepsin K like activity and serum calcium and hydroxyproline concentrations did not change. E-64a inhibited femoral cathepsin K-like activity in tail-suspended rats, but etidronate did not. E-64a as well as etidronate significantly prevented the suspension-induced declines in bone weight and strength. However, more frequent injection and higher doses were required for E-64a to exhibit significant efficacy of antiresorption, compared with those of etidronate. Our results suggest that a cysteine protease inhibitor could improve suspension-induced osteopenia by inhibiting cathepsin K-like activity in bone; however, it needs several improvements in the effect as a clinical drug. PMID- 11938610 TI - Observations on normal body temperatures in Vietnamese and Japanese in Vietnam. AB - The observations described in this paper were made during a study of the effects of tropical climate upon Vietnamese and Japanese. We measured rectal and skin temperatures every 10 min for 26 hrs in 6 Vietnamese and 6 Japanese. The experiments have been conducted for 2 hot months, June and July 1999 and 2000 in Hanoi, Vietnam. The ambient temperatures ranged from 33 to 36 degrees C. The results obtained are summarized as follows: 1) Skin temperatures at thigh, forearm and hand during the daytime were significantly higher in the Vietnamese subjects than in the Japanese ones. It would be physiological reactions to warm ambient temperatures, which is advantageous for dissipation of body heat. 2) The average rectal temperature during the daytime is significantly higher in the Vietnamese than in the Japanese, while significantly lower at night. 3) Average range of oscillation of rectal temperature was 1.26 degrees C in the Vietnamese, which was clearly greater than in the Japanese. The higher core temperature, which was actively regulated under warm temperature, seemed of adaptive significance, resulting in the reduction of water consumption like camels in the desert. A greater range of oscillation in tropical Vietnamese people might have ecological significance for efficient acclimatization in the warm environment, suggesting that the setpoint of core temperature could show a greater range of oscillation. PMID- 11938611 TI - Effects of clothing pressure caused by different types of brassieres on autonomic nervous system activity evaluated by heart rate variability power spectral analysis. AB - The present study was designed to investigate the effects of clothing skin pressures exerted by two different types of brassieres (a conventional higher skin-pressured brassiere and a newly devised low skin-pressured brassiere) on the autonomic nervous system (ANS) activity. Six healthy young women (22.8 +/- 1.4 yrs.) with regular menstrual cycles participated in this study. The ANS activities were assessed by means of heart rate variability power spectral analysis. The skin pressures exerted by the brassieres were measured with an air pack type contact surface pressure sensor at five different points. The total amount of clothing pressure, and the pressures at the center and the side regions of the brassieres were significantly greater in the high than in the low skin pressured brassiere (Total 9816.1 +/- 269.0 vs. 6436.8 +/- 252.4 Pa, P < 0.01; Center 2212.1 +/- 336.3 vs. 353.8 +/- 85.8 Pa, P < 0.01; Side 2556.8 +/- 316.1 vs. 1747.2 +/- 199.2 Pa, P < 0.05). Concerning the ANS activity, the Total power, and the very low frequency (VLF) and the high frequency (HF) components were significantly decreased in the high skin-pressured brassiere than those in the low skin-pressured brassiere (Total 531.6 +/- 57.3 vs. 770.5 +/- 54.2 ms2, P < 0.01; VLF 60.7 +/- 14.6 vs. 179.2 +/- 38.1 ms2, P < 0.05; HF 209.5 +/- 33.2 vs. 283.2 +/- 61.5 ms2, P < 0.01). Our data indicate that the higher clothing pressures exerted by a conventional brassiere have a significant negative impact on the ANS activity, which is predominantly attributable to the significant decrease in the parasympathetic as well as the thermoregulatory sympathetic nerve activities. Since the ANS activity plays an important role in modulating the internal environment in the human body, excess clothing pressures caused by constricting types of foundation garments on the body would consequently undermine women's health. PMID- 11938612 TI - A cooling vest for working comfortably in a moderately hot environment. AB - To alleviate worker's thermal discomfort in a moderately hot environment, a new cooling vest was designed and proposed in this paper. To investigate the effect of the cooling vest and to collect the knowledge for the design of comfortable cooling vest, subjective experiments were conducted. Two kinds of cooling vests, the new one and the commercially available one, were used for comparison. The new cooling vest had more insulation and its surface temperature was higher than the commercially available one. Experiments were performed in the climatic chamber where operative temperature was controlled at 30.2 degrees C and relative humidity was at 37% under still air. In addition, experiment without cooling vest was carried out as a control condition. The results obtained in these experiments were as follow: 1) By wearing both types of cooling vest, the whole body thermal sensation was closer to the neutral conditions than those without cooling vest. This effect was estimated to be equal to the 5.7 degrees C decrement of operative temperature. The subjects felt more comfortable with the cooling vest than without it. They felt more thermally acceptable than that without cooling vest. Wearing the cooling vest was useful to decrease the sweating sensation. 2) The local discomfort was observed when the local thermal sensation was "cool" approximately "cold" with the cooling vest. 3) The new cooling vest kept the skin temperature at chest at about 32.6 degrees C. On the other hand, by wearing the commercially available one, it lowered to about 31.1 degrees C. By wearing the new cooling vest, there was a tendency that local thermal sensation vote was higher and local comfort sensation vote was more comfortable than those of the condition wearing the commercially available one. It is important for the design of a comfortable cooling garment to prevent over-cool down from the body. PMID- 11938613 TI - Leadership challenges in clinical and information technology services. PMID- 11938614 TI - A quick view ... neonatal vs. adult ventilation. PMID- 11938615 TI - The fundamentals of .... ultrasound equipment. PMID- 11938616 TI - A review of.... Unshielded twisted pair and Cat 5. Part 1. PMID- 11938617 TI - Tips on preparing for an exam. PMID- 11938618 TI - 21st century brings new focus and novel methods to health care training. PMID- 11938619 TI - Technology and patient safety: a two-edged sword. AB - The Agency for Healthcare Research and Quality (AHRQ) is now the world's largest funder of patient safety research. Part of AHRQ's research focus is to examine evidence to help determine which technologies can be used to effectively minimize harm and improve patient safety. The report of the Institute of Medicine (IOM). To Err is Human stressed the importance of automating repetitive, time-consuming, and error-prone tasks through the use of technology. While automation holds substantial promise for improved safety, error experts caution that all technology introduces the potential for new and different errors. It is critical that any new automated system be tested in actual operational settings to determine what, if any, unanticipated failures exist. Field-based research is essential in the emerging field of patient safety to create the evidence as to which technologies actually improve patient safety and those that may well increase the potential for harm. PMID- 11938620 TI - Engineering and governmental challenge: 7-day/24-hour chronobiologic blood pressure and heart rate screening: Part I. AB - This review provides evidence that the bioengineering community needs to develop cost-effective, fully unobtrusive, truly ambulatory instrumentation for the surveillance of blood pressure and heart rate. With available instrumentation, we document a disease risk syndrome, circadian blood pressure overswinging (CHAT, short for circadian hyper-amplitude-tension). Circadian hyper-amplitude-tension is defined as a week-long overall increase in the circadian amplitude or otherwise-measured circadian variability of blood pressure above a mapped threshold, corresponding to the upper 95% prediction limit of clinically healthy peers of the corresponding gender and age. A consistently reduced heart rate variability, gauged by a circadian standard deviation below the lower 5% prediction limit of peers of the corresponding gender and age, is an index of a separate yet additive major risk, a deficient heart rate variability (DHRV). The circadian amplitude, a measure of the extent of reproducible variability within a day, is obtained by linear curve-fitting, which yields added parameters: a midline-estimating statistic of rhythm, the MESOR (a time structure or chronome adjusted mean), the circadian acrophase, a measure of timing of overall high values recurring in each cycle, and the amplitudes and acrophases of the 12-hour (and higher order) harmonic(s) of the circadian variation that, with the characteristics of the fundamental 24-hour component, describe the circadian waveform. The MESOR is a more precise and more accurate estimate of location than the arithmetic mean. The major risks associated with CHAT and/or DHRV have been documented by measurements of blood pressure and heart rate at 1-hour or shorter intervals for 48 hours on populations of several hundred people, but these risks are to be assessed in a 7-day/24-hour record in individuals before a physical examination, for the following reasons. (1) The average derived from an around the-clock series of blood pressure measurements, computed as its MESOR, the proven etiopathogenetic factor of catastrophic vascular disease, can be above chronobiologic as well as World Health Organization limits for 5 days or longer and can be satisfactory for months thereafter, as validated by continued automatic monitoring. The MESOR can be interpreted in light of clock-hour-, gender-, and age-specified reference limits and thus can be more reliably estimated with a systematic account of major sources of variability than by casual time-unspecified spot checks (that conventionally are interpreted by a fixed and, thus, rhythm, gender-, and age-ignoring limit). With spot checks, in a diagnostically critical range of "borderline" blood pressures, an inference can depend on the clock-hour of the measurement, usually providing a diagnosis of normotension in the morning and of hypertension in the afternoon (for the same diurnally active, nocturnally resting patient!). Long-term treatment must not be based upon the possibility of an afternoon vs a morning appointment. Moreover, the conventional approach will necessarily miss cases of CHAT that are not accompanied by MESOR hypertension. (2) Circadian hyper-amplitude-tension indicates a greater risk for stroke than does an increase in the around-the-clock average blood pressure (above 130/80 mm Hg) or old age, whereas (3) CHAT can be asymptomatic, as can MESOR hyptertension. (4) Deficient heart rate variability, the fall below a threshold of the circadian standard deviation of heart rate, an entity in its own right, is also a chronome alteration of heart rate variability (CAHRV). Deficient heart rate variability can be present together with CHAT, doubling the relative risk of morbid events. In each case--either combined with CHAT or as an isolated CAHRV--a DHRV constitutes an independent diagnostic assessment provided as a dividend by current blood pressure monitors that should be kept in future instrumentation designs. CHAT and DHRV can be screened by systematic focus on variability, preferably by the use of automatic instrumentation and analyses, which are both available (affordably) for research in actual practice, in conjunction with the Halberg Chronobiology Center at the University of Minnesota. PMID- 11938621 TI - Use of laboratory studies in rheumatologic diseases. PMID- 11938622 TI - Is it gout? Tap the joint! PMID- 11938623 TI - Osteoarthritis--a review. PMID- 11938624 TI - Rheumatoid arthritis. PMID- 11938626 TI - American College of Rheumatology. PMID- 11938625 TI - Antiphospholipid syndrome: diagnosis, pathogenesis, and management. PMID- 11938627 TI - OIG approves plan involving hospital investment in physician-owned ambulatory surgical center. PMID- 11938628 TI - Measuring physician performance. PMID- 11938629 TI - A perspective on arthritis. PMID- 11938630 TI - Theodosia Burr Alston: medical history and mysterious death. PMID- 11938631 TI - Sir William Osler as an athlete: an under-examined facet of his life. PMID- 11938632 TI - Impact of cortical perforations of contiguous donor bone in a guided bone augmentation procedure: an experimental study in the rabbit skull. AB - BACKGROUND: It has been shown that bone can be augmented beyond the original skeletal envelope by using space-making barriers. Further, it has been suggested that perforation of the contiguous donor bone enhances bone formation in guided bone augmentation procedures. PURPOSE: The goal of the present investigation was to evaluate whether perforations into the donor bone marrow through the cortical plate, located contiguous to an extracalvarial experimental space, influence bone generation into this space with regard to augmented bone tissue volume and bone density 3 months postoperatively. MATERIALS AND METHODS: Two titanium cylinders, each with a titanium lid, were subcutaneously placed with their open ends facing the parietal bones of eight rabbits and secured with miniscrews. The cortical bone plate on the test side was perforated with seven evenly distributed holes, each with a diameter of 1.2 mm, using carbon-steel burs. Together, these perforations corresponded to about one-third of the total experimental bone area. The bone on the control side was left intact, and no bleeding occurred during the placement of the titanium lid. The perforation procedure (test side) resulted in various degrees of blood fill. After 3 months, the animals were sacrificed to obtain ground sections for histology and histomorphometry. RESULTS: The cylinders were found to be partly filled with tissue containing slender bone trabeculae and marrow spaces in abundance. The bone consistently reached a higher level at the inner wall compared with the central part of the cylinders (p = .001). Hollow connections between the experimental space and the skull bone marrow were found in the contiguous outer cortical plate in four of the seven control sites. No statistically significant differences could be demonstrated between the perforated test sites and the control sites regarding augmented tissue volume (64.4 +/- 18.9% vs. 64.9 +/- 22.2%) or bone density, although there was a tendency toward denser bone in the test sites (21.5 +/- 11.1%) versus control sites (14.7 +/- 5.4%). There was no statistical difference regarding relative bone-to-titanium wall contact (27.4 +/- 14.7% for test; 38.6 +/- 25.9% for control). Thickness (height) and density of the skull bone vault were measured in the area beneath and lateral to the cylinders. No significant differences could be observed regarding these parameters between the test and control side. There were no correlations between thickness (height) or density of donor bone versus amount or density of augmented bone. The degree of immediate blood fill could not be shown to correlate with augmented tissue volume or augmented bone density. CONCLUSIONS: In the present model, as observed 3 months postoperatively, cortical perforations of contiguous donor bone or degree of immediate blood fill of an extracalvarial experimental space were not found to enhance augmented tissue volume beyond the skeletal envelope. Although there was a much higher mean value for bone density of augmented bone in the test sites, the large variations failed to show significant intergroup differences. PMID- 11938633 TI - Subjective need for implant treatment among middle-aged people in Sweden and Denmark. AB - BACKGROUND: The use of oral implants in prosthodontics has become widespread and regarded as a predictable treatment modality. However, there is a lack of knowledge among the general population about the prevalence and need for implant treatments. PURPOSE: This study was undertaken to register and compare the prevalence of dental implants and the subjective need for implant treatment among people in Sweden and Denmark. MATERIALS AND METHODS: Random samples taken from the national population registers in Sweden and Denmark comprised 1001 Swedish subjects aged 55 to 79 years and 1175 Danish subjects aged 45 to 69 years. Subjects were requested to fill out questionnaires regarding dental conditions, subjective need for implant treatment, whether they had received treatment with dental implants during the previous 10 years, and so on. RESULTS: Of the Swedes, 4.8% reported that they had dental implants, compared with 2.5% of the Danes. In the Swedish sample, age was significantly associated with subjective need for implant treatment. In the Danish sample, women showed a significantly higher subjective need for implant treatment than did men. CONCLUSIONS: Compared with the Swedish sample, the subjective need for treatment with dental implants was higher in the Danish sample, although the patient fees were substantially higher in Denmark. PMID- 11938634 TI - Clinical and radiographic evaluation of the 5-mm diameter regular-platform Branemark fixture: 2- to 5-year follow-up. AB - PURPOSE: The purpose of this study was to report the clinical results obtained with 5-mm diameter regular-platform Branemark implants after 2 to 5 years of follow-up. A secondary aim was to compare the clinical outcome to that of adjacently placed standard 3.75-mm fixtures in the same patients under the same operative condition and in the same prosthetic construction. MATERIALS AND METHODS: Sixty patients, with a mean age of 54 years, received a total of 97.5-mm diameter regular-platform fixtures. The majority of the implants were placed at molar sites. In 41 of the patients, 53.5-mm diameter implants were placed adjacent to 62 standard 3.75-mm diameter fixtures in the same prosthetic reconstruction. All implants were submerged for an average period of 4 to 6 months. Abutment connection was done according to standard protocol. The prosthetic treatment consisted of freestanding fixed bridges. RESULTS: The cumulative survival rate of the 5-mm diameter implants loaded for a period of 2 to 5 years was 96.9%. Only three implants failed. They were placed in type 4 bone in the posterior maxilla. Bone loss over the first year was 0.70 mm and over a 3 year period 0.81 mm. Implants placed in type 4 bone showed significantly higher bone loss. No difference in the resorption rate could be found between the maxillary and the mandibular implants or between the various implant lengths. There was no significant difference between the bone loss around the 5-mm diameter fixtures and the adjacent 3.75-mm diameter standard fixtures. CONCLUSION: The present study demonstrated a high predictability of 5-mm diameter regular-platform implants when placed in the posterior maxilla and mandible. PMID- 11938635 TI - Recent outcomes and perspectives of the application of bone morphogenetic proteins in implant dentistry. AB - BACKGROUND: Since the discovery of bone morphogenetic proteins (BMPs), the number of related studies has increased substantially, and more recent outcomes have cast encouraging perspectives on their use in reconstructive surgery. PURPOSE: The aim of the present review was to summarize the present knowledge about the use of BMPs in conjunction with dental implants based on the literature. MATERIALS AND METHODS: Scientific articles dealing with the use of growth factors and bone healing with or without dental implants were searched for on MEDLINE and critically scrutinized. RESULTS: Thirty-nine scientific reports formed the base for the present review. Whereas the osteoinductive capability of BMPs is well documented, studies on their effects in implant dentistry are still incipient. Preclinical and clinical studies did not show outstandingly good outcomes of the application of BMPs compared with conventional treatments or controls. CONCLUSIONS: The number of studies in the field of dental implantology in which BMPs have been used is still too small for establishing clinical protocols of their use in order to improve a recipient bone bed prior to implant placement or to enhance the integration process of an implant. PMID- 11938636 TI - Early loading of unsplinted implants supporting mandibular overdentures using a one-stage operative procedure with two different implant systems: a 2-year report. AB - BACKGROUND: Step-wise reduction in loading protocols is necessary to evaluate early loading of implants with mandibular overdentures. PURPOSE: To compare the success rates of two different dental implant systems following conventional or early loading protocols in patients being rehabilitated with mandibular overdentures. MATERIALS AND METHODS: Forty-eight edentulous participants were randomly allocated to two different implant systems: one with a machined titanium implant surface (Sterioss, Nobel Biocare, Yorba Linda, California, USA) and the other with a roughened titanium surface (Southern Implants, Irene, South Africa). For each system, the participants were further divided into control groups, in whom mandibular implant overdentures and their respective matrices were inserted following a standard 12-week healing period, and test groups, in whom a 6-week healing period was followed prior to identical loading. Two unsplinted implants to support implant overdentures were placed in the anterior mandible of all participants, using a standardized one-stage surgical procedure. Mobility tests and marginal bone levels, as well as peri-implant parameters, were evaluated at each baseline and 52 and 104 weeks after surgery. RESULTS: There was no statistically significant difference in the success rates of the two systems in either control or test groups. At the 2-year evaluation, a success rate was found of 87.5% and 70.8% for the control and test Sterioss groups, respectively, and 83.3% and 100% for the control and test Southern Implants groups were observed. For the Sterioss groups, eight implants were lost at an early stage: seven in the test group and one in the control group. For the Southern Implants control and test groups, no failures were seen at any time interval. There were no significant differences in marginal bone loss, Periotest values, and peri-implant parameters between implant systems or between any of the control or test groups. CONCLUSIONS: Early loading, with step-wise reductions in loading protocols, of unsplinted machined Sterioss and roughened Southern Implants fixtures with mandibular overdentures is possible for up to 2 years. PMID- 11938637 TI - One-year follow-up of an implant with early radiographic signs of loss of osseointegration: case report. AB - BACKGROUND: Failures in dental implant treatment are classified as early and late and depend on certain complications. Proposed preventions and treatment modalities for late complications have been comprehensively reviewed in the literature. Since early implant failures are characterized by lack of osseointegration, generally, removal of the implant is suggested to prevent extensive bone loss that would further complicate implant placements in future. PURPOSE: This case report presents the controversy between clinical findings and radiographic examinations in evaluating the nature of contact between an implant surface and the bone during the healing period. MATERIALS AND METHODS: In one patient scheduled for implant treatment in both jaws, four maxillary and two mandibular implants showed excellent healing, whereas one mandibular implant showed an extensive radiolucent area indicating early failure after 6 weeks of healing. The latter implant was clinically stable and was not removed. RESULTS: Radiographic evaluation 1 year later showed no sign of peri-implant radiolucency of the mandibular implant as seen after primary healing. CONCLUSION: When the bone-implant interface appears to be compromised radiographically but not clinically, such implants may have a favorable prognosis through a currently unknown biologic host response. PMID- 11938638 TI - Clinical and histologic findings for microimplants placed in one stage and loaded for three months: a case report. AB - BACKGROUND: Implants are placed in either one or two stages. There is an absence of histologic human evidence relating to implant integration after loading. PURPOSE: The purpose of this case report was to present clinical and histologic findings for smooth-surfaced titanium turned microimplants placed in one stage and loaded after healing. MATERIALS AND METHODS: Five one-piece microimplants were placed in a fully edentulous mandible. Three microimplants (tests) were placed in one stage and extended through the keratinized mucosa for 3 mm. Two additional microimplants (controls) were placed even with the mucosa. After 3 months of healing, three test implants were loaded for an additional 3 months. At this time, three loaded implants and one control were removed en bloc. RESULTS: Histologic and histometric evaluations were made. For all specimens, there was excellent bone-to-implant contact. The loaded implants had from two to four exposed threads. Using marginal bone levels as the reference, the highest percentage of bone-to-implant contact was noted with the unloaded control implant (92.2%). One nonaxially loaded implant had 66.9% bone-to-implant contact, whereas the axially loaded implants (n = 2) had 77.8% bone-to-implant contact. CONCLUSIONS: Within the limits of this case report, smooth-surfaced, titanium threaded microimplants placed in one stage and loaded for 3 months demonstrated excellent osseointegration, with varying bone-to-implant contact. The amount of bone-to-implant contact may be related to axial implant loading. PMID- 11938639 TI - Enhanced periodontal response and esthetics of implant-supported bridge by the use of galvanoforming technique: case report. AB - BACKGROUND: Galvanoforming restorations have been placed over the past 15 years successfully. They offer several advantages over alloy restorations, including enhanced response to the periodontal tissues, biocompatibility, and superior esthetics. PURPOSE: The purpose of this report is to show the use of the galvanoforming process in dental implant restorations to transfer the benefits of this technique. MATERIALS AND METHODS: Two standard Branemark fixtures were placed submerged in the lower mandible for the restoration of a three-unit bridge. The impression was taken at fixture level, and two cast individual telescope abutments were inserted. The galvanoforming restoration was seated conventionally without any screw retention. RESULTS: An implant-supported galvanoforming bridge is functioning successfully. The use of biocompatible materials does not compromise the stability of the restoration; instead, the effect on the periodontal tissues is excellent, resulting in less plaque accumulation and bleeding on probing. Microgaps were avoided by conventional seating on the individual telescope gold abutments, revealing superior occlusal esthetics. CONCLUSIONS: This case report demonstrates the practicability of the biocompatible galvanoforming procedure for implant-supported restorations enhancing periodontal response and esthetics. PMID- 11938640 TI - Economy class syndrome and other lessons from postmodern society. PMID- 11938641 TI - Transforming nurses' anger. Interview by Marie Manthey. PMID- 11938642 TI - Healing after a nursing strike. Hearing the voices of nurses. PMID- 11938643 TI - [Architecture of physiological functions: the same basis but new aspects]. AB - The principles of physiological functions formulated by J. Barckroft (constancy of the internal medium, reserves, any adaptation as an integration, principle of antagonism, doubling of mechanisms) are compared with principles of modern physiology. The place and role of physiology in the life sciences are discussed. The necessity of taking into consideration 4 level of regulation of functions (the nervous system, hormones, autacoids, physicochemical factors of the extracellular fluid) is substantiated, as well as the necessity of identification of 4 levels of organization of physiological systems. The main role of the water salt homeostasis in maintaining the cell volume is suggested. Significance of various types of receptors and second messengers in regulation and modulation of functions is shown. PMID- 11938644 TI - [Possible mechanism of cannabinoid-mediated modulation of signal transduction through the basal ganglia]. AB - A possible mechanism of cannabinoid-mediated akinesia is suggested. This effect is proposed to be the consequence of a decrease in LTD/LTP in cortical inputs to striatopallidal/striatonigral cells in the matrix due to CB1 receptor activation. In addition, cannabinoids can attenuate locomotor activity due to a reducing of glutamate/GABA release from axon terminals of subthalamic nucleus/striatonigral cells of matrix and subsequent decrease/increase in the activity of neurons of globus pallidus/substantia nigra pars reticulata. Cannabinoid-mediated rise of dopamine release might be a result of a decrease of dopamine neuron inhibition by striatonigral cells of striosomes. It follows from the suggested mechanism that an inactivation (activation) of CB1 receptors leading to rise (lowering) of the motor activity can be useful for treatment of Parkinson (Huntington) disease. PMID- 11938645 TI - [Recovery of the dog myocardial contractile function in the diastolic period]. AB - Isolated canine heart has an expressed ability for autoregulation of mechanical restitution irrespective of the influence of neurohumoral factors and Frank Starling law on the work of the heart. Mechanical restitution of canine heart in diastolic period starts after the end of mechanical refractory period of the heart and develops exponentially. The higher the heart rhythm the faster the speed of mechanical restitution. The higher the heart rhythm the shorter the mechanical refractory period. Mechanical refractory period of the heart is longer than bioelectrical refractory period. PMID- 11938647 TI - [Modification of redox sites of N-methyl-D-aspartate receptors affects changes in bioelectrical activity of olfactory cortex slices induced by anoxia in rats]. AB - The effects of the thiol oxidative agent 5,5'-dithiobis (2-nitrobenzoic acid) (DTNB) inhibiting NMDA receptors activity on changes of the evoked focal potentials generation (the NMDA and non-NMDA components EPSP) in response to long term (LA) and short-term (SA) anoxic exposure which induced functional disturbances or promoted increasing of neurons resistance to LA, were studied on the rat olfactory cortex slices (Wistar-Kyoto). It was revealed that DTNB (200 mkM) effectively protected the depression of the EPSP generation induced by LA in the most tested neurons. In addition, DTNB eliminates the protective effect of the SA on focal EPSP generation evoked by LA. Nevertheless this dependence concerns the NMDA component and, to a lesser extent, the non-NMDA component EPSP. A possible role of changes of the NMDA receptor modulatory redox sites in mechanisms of functional disturbances and increasing neuronal resistance induced by hypoxic influences, is discussed. PMID- 11938646 TI - [Changes in autoregulation of coronary blood flow in rats with various sensitivity to stress]. AB - The purpose of work: study of a role of endothelial nitric oxide in development of stress-induced changes in autoregulation of coronary blood flow in rats with various types of behaviour. The experiments were performed on isolated hearts of female-rats, in the "open field" test, depending on the type of impelling and searching activity of animals subdivided into two groups: "active" and "passive". After a 6-hour immobilization stress only in "passive" rats an increase of volumetric velocity of coronary flow; a decrease of an autoregulation index, coronary reserve against the background of intravascular pressure reduction, were found out. The blockade of nitric oxide synthesis in this group completely eliminated the stress-induced decrease of coronary vascular tone and essentially limited the caused by stress dissociation of coronary flow and the contractility function of the myocardium. In blood plasma of "passive" animals the nitrite/nitrate contents was by 55% more than of the "active" rats. After the transferred stress, in "passive" animals the nitrite/nitrate concentration in blood plasma increased by 29% and in "active" rats--by 136%; the absolute values, however, did not differ between the groups. Thus the autoregulation of coronary flow seems to be subject to action of stress in the rats showing a "passive" type of behaviour in the test "open field", and practically does not change in "active" animals; secondly, in spite of the fact that the stress-induced amplification of NO-producing function andothelium of coronary blood vessels is stereotyped in different rats, in "passive" rats, apparently, sensitivity of coronary vessels to nitric oxide is higher than at "active" those. PMID- 11938648 TI - [Effect of antenatal and postnatal hypoxia on the central nervous system and its correction with peptide hormones]. AB - Ante- and postnatal acute hypoxia significantly aggravated the postnatal development. The posthypoxic behaviour patterns included hyperactivity and training ability inhibition typical for the attention deficit syndrome. A preventive injection of peptide constellation significantly improved the posthypoxic postnatal development and abolished the most of the negative modifications of behavioural patterns. PMID- 11938649 TI - [Mediator secretion in the nerve-muscle synapse in the frog following long-term effect of calcium-free solutions]. AB - The changes of spontaneous and evoked transmitter release in condition of long time (1-4 hours) incubation in Ca-free solution with EGTA adding, were investigated with extracellular recordings in experiments on the nerve-muscular junction of the frog cutaneous-pectoris muscle. Using the method of three extracellular microelectrodes recordings of the monoquantal postsynaptic signals, it was shown that during action of Ca-free solutions the topography of transmitter release changed, the specific spatial organization of points of transmitter release was disrupted. These changes remained after returning to the initial solution. The obtained data suggest that the Ca2+ free solution leads to disruption of active zones of nerve ending. In condition of low initial extracellular Ca2+ concentrations (0.15-0.4 mmol/l), the active zones disorganization led to decreasing of average amplitude of the end-plate currents (EPC) by decreasing their quantal content, increasing their time-course and decreasing the frequency of the miniature end-plate currents (MEPC). The sharp displacement of dependence of quantal contents of EPC in extracellular Ca2+ concentration to a higher Ca2+ concentration without significant changes of slope was revealed. In condition of high (1.8 mmol/l) concentration of Ca2+, the long action of Ca-free solutions leads to decreasing of amplitude of EPC too, but it was less obvious than in condition of initial low Ca2+ concentration. It is supposed that intra- and extracellular Ca concentration provides the support of the typical morpho-functional organization of the mechanisms of transmitter release at the nerve ending of the frog. The disorganization of active zones leads to separation of the elements, which take part at the transmitter release process and reduces the efficiency of secretion. PMID- 11938650 TI - [Electrophysiological study of 5,6-dihydroxytryptamine effect on defensive reflex conditioning in the snail]. AB - The role of serotonin in expression of membrane properties of identified neurons was studied during defensive reflex conditioning using the neurotoxic analogue of serotonin 5,6-dihydroxytryptamine (5,6-DHT). The defensive reflex conditioning in snails was destroyed on the second day after second injection of 5,6-DHT. Through the 1st weeks after second injection of 5,6-DHT the snails were learned but worse than snails after injection of saline solution. This result shows the recovery of snail's learning ability within 2 weeks after the second injection of 5,6-DHT. It was found that injection of 5,6-DHT prevented the decrease of membrane and threshold potentials of command neurons during defensive reflex conditioning as compared with the snails injected with 5,6-DHT without learning. PMID- 11938651 TI - [Dopaminergic processes in the striatum mediating corticoliberin effect on behavior of active and passive rats]. AB - The action of intranasal corticotropin-releasing hormone (CRH) administration on open field behavior and striatal and hypothalamic levels of dopamine, noradrenaline and their metabolites has been studied in rats with different behavior strategies (KHA and KLA strains). In KLA rats, CRH administration resulted in increased locomotor and exploratory activity, while KHA rats demonstrated decreased that. The analysis of catecholamine levels did not detect any strain differences in hypothalamus, but in striatum the dopamine levels have been twice higher, while the metabolite levels (DOPAC and HVA) were significantly lower in KLA rats as compared to KHA rats. The CRH administration led to increased dopamine and noradrenaline levels in hypothalamus and decreased those in striatum in rats of both strains, but in KLA the decrease was more evident. It is probably a result of intensified mediator turnover induced by the neurohormone in KLA rats, as supported by a fact of increased dopamine metabolite levels in this structure. PMID- 11938652 TI - [Electromyographic analysis of the role of the otolith apparatus in regulation of postural disorders in humans with extrapyramidal pathology]. AB - In 30 patients with cervical dystonia in the posture of orthostasis, an obvious change of the electromyographic pattern was shown both at rest and in different functional tests. The analysis has shown an enhanced excitability of motoneurons and a disorder of the cortical control, as well as enhancement of the irradiation process between paired structures controlling activity of opposite muscular groups. After treatment with intratympanal administration of an ototoxical preparation on the opposite side in respect to head turn direction, normalising of the cortical control and diminishing of the excitation irradiation. The data obtained suggest a significant role of the vestibular otolithic apparatus in regulation of postural disorders in extrapyramidal pathology. PMID- 11938653 TI - [The nature of very low frequency component of the heart rate variability and the role of sympathetic-parasympathetic interactions]. AB - A method of separate monitoring "instant" changes of the VLF, LF and HF power spectral components of heart rate variability, has been developed. The power of the LF and HF spectral components were proved to be continuously changing. The period of these power fluctuations could stay within 15 to 150 sec. Comparison of the heart rate variability spectrum with LF and HF spectral components power fluctuations' spectrums has shown that the frequencies of the LF and HF spectral components power fluctuations stay within the VLF range. The co-operative spectrum form of these fluctuations repeats the form of the VLF peak. In cases when the LF and HF spectral components power fluctuations' periods do not coincide, VLF has two peaks. The frequency of one VLF peak coincides with frequency of the HF power fluctuations, and the frequency of another--with the frequency of LF power fluctuations. PMID- 11938654 TI - [Spectral characteristics of electrical activity of the respiratory center in brain processes in fetus and newborn rats in vitro]. AB - Power spectral analysis of inspiratory discharges of C3-C5 ventral roots in brainstem-spinal cord preparation from foetal (18 and 20 gestation days) and newborn (0-1 and 2-3 postnatal days) rats was performed. The respiratory centre perinatal development manifests itself by decreasing of respiratory rhythm variability and increasing of inspiratory burst duration. In foetal inspiratory bursts, low-frequency oscillations (1-10 Hz) dominate. In early postnatal stage, the relative power of low-frequency oscillations begin to decrease, and medium frequency oscillations (10-50 Hz) start to dominate over the inspiratory discharge. The data obtained suggests, that perinatal maturation of respiratory centre is characterised by stabilisation of the respiratory rhythm generation and developmental alteration of inspiratory activity's spectral and temporary parameters. PMID- 11938655 TI - [Human respiratory sensory system, its physiological role]. AB - Modern data on genesis and functional role of sensations associated with respiration, were analysed. Regularities of human perception of the respiratory cycle's parameters have been cited. Respiratory discomfort occurring in an increased loading of the respiratory system, as well as an imperative respiration stimulus developing in a prolonged apnoe or obvious hypoventilation, are regarded as defensive responses to a threat to normal ventilation of the lungs. The central link in mechanisms of the respiratory sensory system seems to involve pulses from receptors of the respiratory muscles. PMID- 11938657 TI - [Oxygen consumption by isolated chorioallantois during later stages of the chick embryo development]. PMID- 11938656 TI - [Changes in the hippocampus excitability following disruption of the amygdala hypothalamus connections]. PMID- 11938658 TI - Effect of carotene on the photovoltaic and optical properties of Langmuir films of chlorophyll. PMID- 11938659 TI - Phage antibodies neutralize vaccinia virus. PMID- 11938660 TI - Comparative analysis of the superoxide dismutase activity in tissues of higher vertebrates. PMID- 11938661 TI - An immunosensor based on Langmuir-Blodgett films and infrared fluorescence detection. PMID- 11938662 TI - Catalytic properties of cholinesterases from the brain and blood serum of mink (Mustela vison Bris.). PMID- 11938663 TI - Fragment E of fibrin as a fibrinogen cofactor in the reaction of enzymatic cross linking. PMID- 11938664 TI - The kinetic mechanism of formate dehydrogenase from bakery yeast. PMID- 11938666 TI - The immunostimulant effect of ubiquinone 50 on the development of viral resistance in plants of the family Solanaceae juss. PMID- 11938665 TI - The role of extracellular DNA in the stability and variability of cell genomes. PMID- 11938667 TI - Computer simulation of the three-dimensional structure of the glutamate site of the NR2B subunit of the NMDA receptor. PMID- 11938669 TI - Estimation of toxicity of monosubstituted phenols. PMID- 11938668 TI - High-temperature stress-related changes in the harmonics F0, Fm, and Fv of pulse amplitude modulated fluorescence signals: locating thermal damage in reaction centers of photosystem II. PMID- 11938670 TI - Energy transfer from tryptophane amino acid residues to retinal in a bacteriorhodopsin molecule within a femtosecond timescale. PMID- 11938671 TI - A new scheme of lignin biosynthesis and the mechanism of its regulation of functional properties. PMID- 11938672 TI - Thermostable DNA polymerase from the archaeon Archaeoglobus fulgidus. PMID- 11938674 TI - Effect of chloride and bicarbonate on the plasma membrane Mg(2+)-ATPase of bream (Abramis brama L.) brain, which is sensitive to the inhibitor receptor ligands. PMID- 11938673 TI - Inhibition of HIV-1 reverse transcriptase by aryl-substituted naphto- and anthraquinones. PMID- 11938675 TI - Some spectral characteristics of pigment-protein complexes and their interaction in membranes of Thiorhodospira sibirica. PMID- 11938676 TI - A spatial model of the glycine site of the NR1 subunit of NMDA-receptor and ligand docking. PMID- 11938677 TI - [Nosocomial infections in pediatric and neonatal intensive care: an epidemiological update]. AB - Hospital-acquired infection (nosocomial infection) is a world-wide problem. The peculiar susceptibility of newborns and children, the widespread use of antibiotics, advances in hospital practice, have resulted in an increased incidence and recognition of neonatal and pediatric bacterial nosocomial infections. The aim of this review is to present an update on epidemiology of hospital-acquired infections, quoting recent and relevant papers on this topic. Main risk factors for the neonatal and the pediatric intensive care infections are presented, respectively. Emerging antibiotic-resistant gram-positive and gram negative bacteria in intensive care units are also considered. Infection control programmes are necessary to monitor and to prevent nosocomial infections. PMID- 11938678 TI - [Congenital and acquired chylothorax]. AB - BACKGROUND/PURPOSE: Chylothorax in paediatric age is a life-threatening clinical entity that cause serious respiratory, nutritional and immunologic complications. Chylothorax in the absence of trauma or tumour is uncommon and lymphangiomatosis of the bone, although extremely rare, has been associated with these condition. The authors describe the case of a two-year-old girl who presented with a massive chylothorax associated with hip and paravertebral lymphangioma and spread lymphangiomatosis of the spine. The authors also review the literature and their experience of congenital and postoperative chylothorax in order to establish guidelines for the diagnosis and management of both primary and postoperative chylothorax in paediatric age. METHODS: From 1990 and 1999, 14 children had chylothorax. 9 patients had pleural effusion after surgical procedure, 5 patients had congenital chylothorax (both in prenatal and neonatal time), one of whom with bone lymphangiomatosis associated. RESULTS: Postoperative chylothorax has been successfully treated by conservative approach (starvation, total parenteral nutrition and chest tube) in 6 out of 7 cases (two patients died because of complex cardiac malformation). Conservative approach is useful in case of congenital chylothorax, but not with bone lymphangiomatosis associated. CONCLUSIONS: Postoperative and congenital chylothorax is well managed with conservative treatment. Chylothorax with bone lymphangiomatosis associated needs early and aggressive surgical approach. PMID- 11938679 TI - [Characteristics of 86 infants' cluster with A.L.T.E. (Apparent Life-Threatening Event): similarities and differences versus main risk factors for SIDS]. AB - In this study we have gathered 86 ALTE infants' characteristics about physiological, familiar, remote and near pathological case history, modality of apnoea episode's presentation, environmental and behavioural conditions of babies and we have compared these ones with main risk factors for SIDS to find possible similarities or differences. Many of the already known differences or similarities between ALTE and SIDS have been confirmed by our data and new ones have been observed. So we can say that the association between ALTE and SIDS remains contrasting because beside undiscussed factors of association there are other ones which differentiate the two Syndromes. PMID- 11938680 TI - The nonpalpable testis: an experience of 132 consecutive videolaparoscopic explorations in 6 years. AB - From may 1995 to may 2001, 114 children with nonpalpable testis (NPT) were evaluated at our institution (18 babies had bilateral cryptorchidism). The age range was 1-11 years. When ultrasonography and nuclear magnetic resonance cannot show the position of the NPT along the normal pathway, video laparoscopy is essential for diagnostic accuracy. 20 cases were observed to have blind-ending was deferens and testicular vessels; therefore, no other procedure was done. 4 had residual nonfunctional intra-abdominal tissue; in 52 cases, an intra abdominal testis was found, and 17 microvascular and 26 traditional orchidopexies were performed. The remaining 50 patients (six with bilateral cryptorchidism) had normal vas and spermatic vessels entering the inguinal canal. In 15 cases a normal testis was present, and it was positioned into the scrotum with the standard technique; in 41 cases an atrophic testis was found and was removed through an inguinal approach. Diagnostic laparoscopy permits not only localisation of the testis but also planning for a better therapeutic program with a minimally invasive procedure, thus avoiding the knife in 18% of cases (in our experience 15% of blind-ending and 3% of abdominal vanishing testis). PMID- 11938681 TI - The role of endoscopic surgery in paediatric oncological diseases. AB - Recent improvements in video imaging and instrumentation have encouraged a wider use of endoscopic surgery as a modality for diagnostic and operative procedures. To asses the utility and diagnostic accuracy of endoscopic surgery in children with oncological diseases, we reviewed our experience about thirty-eight patients affected by oncological diseases, referred to our Department since 1995. We performed laparoscopy in 22 cases, thoracoscopy in 14, 1 combinated procedure (laparoscopy + thoracoscopy) and retroperitoneoscopy in 1 case. Endoscopic surgery is indicated in cases of paediatric oncological diseases both for diagnosis and treatment. PMID- 11938682 TI - [Alimentary strategies in the neonatal period in the prevention of allergies]. AB - The Authors report an update relative to the dietetic prevention strategies in the high allergic risk subject, as proposed from more recent literature. The babies with a familiar history of atopia are defined as population with allergic risk. The Authors examine the role of early exposure to cow's milk formulas and maternal diet during breast-feeding as risk factors for allergic symptoms in such babies. Moreover, they examine the indications for hydrolisated milk (partial and extensive) formulas and soy milk formulas use, as reported in published Meta analysis and official statements of several Scientific Associations. They conclude that beyond the undoubted preventive role of exclusive breast-feeding in the first 4-6 month after birth, and of the extensively hydrolisated formulas, there are many concerns about the role for partially hydrolisated formulas and soy formulas. The Authors claim for multicentric methodologically correct trials in order to clear the controversies. PMID- 11938683 TI - [Prophylactic total thyroidectomy in children and adolescents with genetic mutations in the RET-protooncogene]. AB - Medullary thyroid cancer (C.M.T.) can be a sporadic form generally in adults or a heredofamilial form where the first symptom appears in pediatric and adolescent age. The hereditary form can be isolated or associated with others endocrine neoplasias of type 2: MEN2a (with or without cutaneous lichen amyloidosis) and MEN2b. The responsible gene of the transmission has been identified in proto oncogene RET localized on chromosome 10. Point form mutations of this proto oncogene have been found on exons 10 and 11 in MEN2a and on 16 in MEN2b. In our study on 64 subjects, who belong 11 familiar groups, affected by MEN2a, MEN2b and familiar C.M.T., underwent a genetic research to look for point form mutations of proto-oncogene RET with PCR followed by the analysis of restriction. A genetic mutation has been revealed in 25 subjects: 18 were already known affected by MEN2 and so surgical treated and 7 seemed healthy (mean age 17.4 years, range 10-25). These 7 patients has been undergone clinical research and surgical treatment: a total thyroidectomy associated a lymphectomy of the central compartment. In all cases the histological exam showed C.M.T. moreover a patient had metastasis in lymph nodes of the central compartment. Another had hyperparathyroidism and pheochromocytoma treated with total thyroidectomy, parathyroidectomy and bilateral laparoscopic adrenalectomy. The identification in a very early age of carrier subjects of hill's gene inside an affected family, permits the execution of a prophylactic total thyroidectomy to prevent the C.M.T.. The penetrance of this neoplasia in hereditary form is 100%. PMID- 11938684 TI - Laparoscopic decapsulation of a large epidermoid splenic cyst in a child using the UltraCision LaparoSonic Coagulating Shears. AB - Splenic cysts are rare in pediatric surgery. Congenital epidermoid cysts are exceptional representing only 2.5% of all splenic cysts in childhood. Nowadays, considering the short- and long-term complications of splenectomy in children, the management of epidermoid cyst consists of partial splenectomy or decapsulation of the cystic wall. To our knowledge, the case reported in this article describes the first successful laparoscopic decapsulation of an epidermoid splenic cyst in an 10-year-old child using the UltraCision LaparoSonic Coagulating Shears (LCS). Follow-up at six months confirms no recurrence. Laparoscopic splenic decapsulation provides minimal access and small surgical trauma for treating the cyst while preserving splenic function. The use of UltraCision LCS makes the laparoscopy safely, expeditiously, with minimal blood loss and short hospital stay. PMID- 11938685 TI - Congenital true pancreatic cyst in pediatric age: case report. AB - Congenital pancreatic cysts are rare lesions in pediatric age. We present a case of congenital true pancreatic cyst in otherwise asymptomatic boy aged 4 years. Clinical history, laboratory, imaging, immunohistological findings and surgical treatment are also described. PMID- 11938686 TI - [Pica and iron-deficiency anemia in a 2-year old Indian child]. AB - Pica is the compulsive ingestion of nonnutritive substances. The causes are not known, but the symptom is often associated either with iron-deficiency or with pregnancy. The authors report on a case of pica in a 2-year old Indian child, affected by a serious ferropenic anaemia. PMID- 11938687 TI - Dr. Spock: reversing the trend. PMID- 11938688 TI - [Isolated ovarian torsion in a 4-year-old girl: diagnostic setting and laparoscopic treatment]. AB - Authors report a 4 years old girl with isolated torsion of the right ovary and describe diagnostic management (US, CT and MR imaging) and laparoscopic approach. PMID- 11938689 TI - [Catch-up growth and final height in celiac disease]. AB - BACKGROUND: Growth retardation resulting from celiac disease diagnosed in infancy or in early childhood is remediable after introduction of a gluten-free diet. The aim of this study was to evaluate long-term growth, catch-up growth and final height in a group of treated patients with celiac disease. METHODS: The study population consisted of 35 patients (13 M, 22 F) with typical celiac disease. Age at diagnosis was 1.17 (+/- 0.69) years. For each patient, the standard deviation score (SDS) and centile of height at diagnosis, target height and final height were elaborated, and linear regression and correlations between SDS of final height and age at diagnosis, SDS of height at diagnosis and SDS of target height, respectively, were calculated. RESULTS: At diagnosis patients had a general tendency to short stature: mean height was 81.8 cm +/- 22.9 cm (SDS -0.75 +/- 1.61), while target height was 164.3 cm +/- 13.5 cm (SDS -0.14 +/- 1.04) and final height 169.2 cm +/- 7.7 cm (SDS 0.41 +/- 1.04). Linear regression and correlations between SDS of final height and age at diagnosis, SDS of height at diagnosis and SDS of target height, respectively, proved nonsignificant. CONCLUSIONS: Irrespective of age and height at diagnosis, catch-up growth was complete in this group of patients, with satisfactory achievement of final height. Only one patient (age at diagnosis 13 years) failed to reach the target height. PMID- 11938690 TI - Two new species of the genus Syringophilopsis Kethley, 1970 (Acari: Syringophilidae) parasitizing quills of true shrikes (Aves: Laniidae). AB - Two new species of syringophilid mites (Acari: Syringophilidae) are described from quills of true shrikes (Passeriformes: Laniidae): Syringophilopsis kristini sp. n. from Lesser Grey Shrike, Lanius minor, from Slovakia, and Syringophiopsis yosefi sp. n. from Lanius sp. from Cameroun. PMID- 11938692 TI - Trichostrongylina (Nematoda) from Malagasy Muridae. I--Description of two new species of Heligmonellidae in Nesomys spp. AB - Two new species of Heligmonellidae, Heligmonina wrightae n.sp. (Nippostrongylinae) and Nesomystrongylus fissicauda n.gen., n. sp. (Brevistriatinae) are described from Madagascar in Nesomys rufus and N. audeberti (Muridae). In Nesomys audeberti, the species are coparasites. Heligmonina wrightae is differentiated from all the other species of the genus, except H. malacomysi Sakka & Durette-Desset, 1988, by the ratio of the length of the spicules on the length of the body (25-27.8% versus 9.5-7%). It differs from H. malacomysi by the pattern of the caudal bursa and by the angle of the axis of orientation of the cuticular ridges on the sagittal axis. Nesomystrongylus fissicauda is related to the genus Fissicauda Durette-Desset & Krishnasamy, 1976, by the absence of the carene, by the ridges discontinuous on all the sides of the body and by the deeply divided dorsal ray. It differs from this genus by a different structure of the ridges, by the pattern of the caudal bursa, (very tiny rays 2 and strongly developed rays 3, rays 8 arising from common trunk of rays 2 to 6) and by the presence of a caudal tip in the female. PMID- 11938691 TI - Hepatozoon kisrae n. sp. infecting the lizard Agama stellio is transmitted by the tick Hyalomma cf. aegyptium. AB - Hepatozoon kisrae n. sp. was found infecting a starred lizard at a site in southeastern Samaria, Palestine. These lizards were also hosts to the ixodid tick Hyalomma cf. aegyptium, which was demonstrated to be the vector of this hemogregarine. Hepatozoon and tick infections occurred in lizards within a very restricted locality; at a second site, nearby, ticks occurred without Hepatozoon infection. Micro- and macromeronts occurred mainly in the lungs, while cyst-like merogonic stages, mainly dizoic, occurred in the liver. Mature intraerythrocytic gametocytes were stout and encapsulated. Development from oocysts to sporocysts took place in the tick hemocoel, and was examined by transmission electron microscopy. Lizards were successfully infected when fed on sporocyst-infected ticks or viscera of infected lizards. Ticks become infected when fed on infected lizards; sporogony was complete when the ticks reached adult stage, over 40 days after initial attachment. PMID- 11938693 TI - [The plague in Vietnam: history and inventory of collected fleas (insecta, Siphonaptera) in the inhabited zones]. AB - Ten flea species are reported in anthropic zones of Vietnam. Xenopsylla vexabilis is also here included because of it has been involved in others plague's countries. Lentistivalius klossi is the only selvatic flea known as parasite of synanthropic rats. L. klossi bispiniformis (Li and Wang, 1958), first describe from Chinese specimens, is here synonymized (syn. nov.) with the nominal subspecies. PMID- 11938694 TI - Phylogenetic relationships between the six superoxide dismutase proteins (FeSOD) of Trichomonas vaginalis and FeSOD6 genetic diversity. AB - The parasitic protozoan Trichomonas vaginalis is known to contain several types of Fe-containing superoxide dismutase proteins (FeSOD). Using three different methods of phylogenetic analysis, maximum parsimony (MP), neighbor joining (NJ), and maximum likelihood (ML) methods, we examined the phylogenetic relationships among the six FeSOD (FeSOD1-FeSOD6) based on their amino acid sequences. All the analyses consistently suggested that the six proteins formed a monophyletic group implying that they probably be originated from an ancestral protein form through repeated duplication events. Although MP tree was totally unresolved, the NJ and ML trees revealed that FeSOD6 placed the most basal position and thus emerged earlier than the other five gene types during the evolution of T. vaginalis. Phylogenetic relationships among the five remaining proteins were (FeSOD2, FeSOD3), (FeSOD4, (FeSOD1, FeSOD5)) although weakly supported in terms of bootstrapping values. In addition to this, we newly designed two PCR primer specifically amplifying full-length FeSOD6 gene and examined its genetic diversity among 12 T. vaginalis isolates from five countries and three continents. They had the same nucleotide sequences except those of three Korean isolates which showed one to three different nucleotides. PMID- 11938695 TI - An outbreak of cutaneous leishmaniasis among soldiers in Belem, Para State, Brazil, caused by Leishmania (Viannia) lindenbergi n. sp. A new leishmanial parasite of man in the Amazon region. AB - Eight cases of cutaneous leishmaniasis are recorded among soldiers of the Brazilian Forest Infantry stationed in Belem, State of Para, north of Brazil. The infections, all acquired during manoeuvres in nearby degraded primary forest, are attributed to a new member of the subgenus Viannia, Leishmania (V.) lindenbergi n. sp. A further infection by this parasite was encountered in a woman, who lived very close to the same piece of forest. The new parasite has been characterised and differentiated from other known species of the subgenus Viannia following the combined use of enzyme electrophoresis and monoclonal antibodies techniques. The eco-epidemiology of L. (V.) lindenbergi is discussed: by far the most abundant anthropophilic sandfly in the type locality was identified as Lutzomyia (Nyssomyia) antunesi (Coutinho), and this remains high on the list of possible vectors. PMID- 11938696 TI - Changes in schizogony and drug response in two lines of rodent Plasmodium, P. berghei NK 65 and P. berghei ANKA. AB - White mice were infected with two strains, ANKA and NK 65, of Plasmodium berghei. The parasites were subjected to chloroquine pressure (60 mg/kg at each passage) during 20 passages. We then compared the behaviour of the strains as they acquired chemoresistance. The drug resistance was estimated by the 2% delay time test (D2%), and the schizogonic rhythm by the synchronicity index (SI). Before drug pressure, the ANKA strain had a D2% of 4.34, and a SI of 0.2. This strain became highly drug resistant, but synchronicity increased: the D2% was 2.93, and the SI was 0.36 at the 20th passage. The NK 65 strain had an initial D2% of 4.12, and an SI of 0.2. The chemoresistance acquired during 20 passages was very irregular for this strain: after drug pressure, the D2% was 2.03 and the SI was 0.28. Drug pressure was then removed (for both strains), for 10 passages (no chloroquine). Resistance and synchronicity returned to their initial values. The two strains behaved very differently, in terms of their affinity for reticulocytes, and with chloroquine activity which favours an increase in SI because only merozoites are preserved. PMID- 11938697 TI - Effect of a single dose (600 mg) of albendazole on Loa loa microfilaraemia. AB - The problem of Loa-encephalopathy, which may occur after ivermectin treatment of patients harbouring high Loa microfilarial loads, might be solved if one could find a treatment regimen bringing about a significant but progressive decrease in the Loa microfilaraemia. A trial was performed in Central Cameroon, whose aim was to follow up for 10 months, and to compare the changes in the Loa microfilarial loads in two groups of patients, one treated with a single dose (600 mg) of albendazole (Alben, SmithKline Beecham) given with fatty food, and the other treated with mebendazole (100 mg, twice a day, generic tablets) at a fasting state. The microfilarial loads remained stable in the mebendazole group, whereas a significant decrease in microfilaraemia was recorded in the albendazole group (initial median load: 230 microfilariae per 50 microliters; median load ten months after: 84 microfilariae per 50 microliters). This should encourage further trials to evaluate the effects and the safety of two- or three-day albendazole regimens in patients infected with Loa loa. PMID- 11938698 TI - PCR-RFLP analysis of Giardia intestinalis using a Giardia-specific gene, GLORF C4. AB - A cDNA clone encoding GLORF-C4 was isolated from the WB strain, an assemblage A Giardia intestinalis. Interestingly, GLORF-C4 has been previously reported as an assemblage B-specific gene. Using two primers based on GLORF-C4 of the GS strain, a prototype assemblage B, GLORF-C4 gene was amplified from all the groups of G. intestinalis, and applied to detect the presence of cysts of G. intestinalis from faecal samples of cyst-passers. RFLP analysis of this PCR product successfully classified G. intestinalis into two distinct groups, assemblages A and B. PMID- 11938699 TI - Scanning electron microscopy of Spinochordodes tellinii (Camerano, 1888), (Gordiacea, Nematomorpha). AB - There are many species of Nematomorpha which are deficiently described and therefore pose doubts about their actual taxonomic position. This is the case with Spinochordodes tellinii (Comerano, 1888), which was transferred to four different genera and has been recently considered as species incertae sedis. A female of Spinochordodes tellinii is redescribed in this work under light microscopy and Scanning Electron Microscopy. Cuticle details, shapes and areolar distribution and the features as well as the location of spiniform structures are analysed. The systematic position is discussed. PMID- 11938700 TI - Host age influence on the intensity of experimental Trichuris suis infection in pigs. AB - The impact of age-related resistance on the regulation of population dynamics of adult Trichuris suis was investigated in an experimental pig model. Helminth naive pigs varying in age from five weeks to four years were infected with T. suis to determine susceptibility to infection. Sows had a significantly lower establishment of adult T. suis worms compared with weaner pigs. Adult worm populations were highly overdispersed in both sows and grower pigs contrasted by a more even distribution among weaner pigs. Sows had significantly lower worm fecundities compared to weaner and grower pigs; T. suis from grower pigs, in turn, had reduced fecundity compared to worms in weaner pigs. In conclusion, we provide the first controlled experimental evidence that age-related resistance to T. suis occurs in pigs. PMID- 11938701 TI - Occurrence and maturation of Rhabdochona gnedini (Nematoda: Rhabdochonidae) in the barbels of the Sousa River, Portugal. AB - Data on the seasonal occurrence and maturation of Rhabdochona gnedini Skrjabin, 1946 in its definitive host, the barbel Barbus bocagei, from the Sousa River, in northern Portugal, are presented. Monthly samples were taken from November, 1998 to October, 1999. A total of 178 fish specimens were inspected. R. gnedini occurred with high prevalences during whole year, attaining the highest values of intensity in summer (overall prevalence and mean intensity 85% and 31, respectively). A clear annual cycle of maturation was observed; females with mature eggs occurred in spring and summer. Unidentified Rhabdochona larvae were recorded from the trichopteran larvae Hydropsyche sp. (prevalence and intensity 29% and one to 34, respectively). These larvae are presumptive intermediate host for R. gnedini. PMID- 11938702 TI - Is the Octomacridae the sister family of the Diplozodae? AB - Diplozoidae and Octomacridae are usually considered as sister families. Essentially this is because they are the only polyopisthocotyleans parasitising primary freshwater teleosts. Because of the lack of phylogenetically informative morphological characters to explore the pattern of colonisation of the primary continental freshwater teleosts and in order to understand the appearance of the "natural parabiosis" of Diplozoidae, a molecular phylogeny was inferred by comparing newly obtained partial 28S and 18S rDNA gene sequences of Eudiplozoon nipponicum and Diplozoon homoion with other already available sequences. The phylogenetic analysis seems to show that Diplozoidae and Octomacridae are not sister groups. Thus, the colonisation of primary freshwater teleosts by these two families could be independent. PMID- 11938703 TI - [Human bothriocephaliasis is always present in France (Survey conducted in Haute Savoie between 1993 and 2000)]. PMID- 11938704 TI - [Semont maneuver vs. particle repositioning maneuver: comparative study]. AB - In the present study 80 consecutive cases of PPV of the posterior semicircular canal are presented, with the purpose to evaluate the effectiveness of the treatment and to analyze the secondary Ny and re-test as indicators of the effectiveness of the therapeutic maneuver. The patients were treated in 40 cases with Particle Repositioning Maneuver (PRM) and in the other 40 with the Semont Maneuver, in an attempt to determine the secondary Nystagmus induced during treatment and any residual Paroxysmal Positional Nystagmus (PPNy) upon repetition of the Hallpike positioning after 15 minutes (re-test). Recovery was achieved in 80% of cases after the first session and in 95% of cases with the third session. PRM seemed to be slightly more effective. Secondary liberating Ny was frequently evoked (80%) and was reliable (75%) with the Semont Maneuver, while PRM yielded the liberating Ny in only 25% of cases, with lower reliability (70%). Reliability of the re-test with the Hallpike maneuver was high (80%) and the difference between the two techniques was negligible (78% in PRM and 85% in Semont). In conclusion, in terms of indicators of effectiveness, the Semont liberating Maneuver appears better in that it combines high therapeutic effectiveness (75% recovery in a single session) with a like percentage of liberating Ny (80%): this makes it possible to reserve the re-test and repetition of the therapeutic maneuver, performed during the same session, only in those cases that do not present liberating Ny (20% of cases). On the other hand, when the PRM is used, it appears better to rely on the high likelihood of effectiveness (85% in a single session) and then review the case directly at the next check-up rather than perform the Hallpike re-test in all cases that did not present the liberating secondary Ny (75%). In this way it is possible to select a small number of patients (22%) that still show PPNy (positive re-test) and for whom the therapeutic maneuver must be repeated. PMID- 11938705 TI - [Sign-report: torsional nystagmus from head-shaking in patients with labyrinth lithiasis of the vertical canal]. AB - The clinical picture of Positional Paroxysmal Vertigo (PPV) induced by vertical canal labyrintholithiasis has been clearly described, eliminating previous interpretations of pathogenesis of this disorder. The diagnosis of PPV is based on the well-known picture of positional paroxysmal Nystagmus. The Authors report a clinical sign which has not previously been reported in the literature: torsional Nystagmus induced by the Head-Shaking Test (HST). The Authors encoutered this sign in 30% of the cases of vertical canalolithiasis and in 50% of the cases diagnosed as vertical cupulolithiasis. This sign was also found in patients with a history of prior positional vertigo and in patients who, after treatment with release maneuver, no longer show clinical signs of positional vertigo. Such Nystagmus was not, however, found in the control group (normal subjects and patients suffering from other vestibular pathologies). In the present study the possible pathogenesis of this sign is discussed and some practical implications are considered. PMID- 11938706 TI - [Surgical treatment of neck lymph nodes in squamous cell carcinoma of the pyriform sinus]. AB - The present study reports the results of 66 patients surgically treated for squamous cell carcinoma of the pyriform sinus between 1984 and 1996. Twenty eight patients underwent mono!ateral neck dissection and bilateral neck dissection was performed in 38 cases, for a total of 104 radical neck dissections. Of these, 73 (71%) were modified type III dissections, 17 (16%) were classical, and 14 (13%) were modified type I and II dissections. The primary lesion was strictly lateralized in 47 cases (71%), while median structures were involved in 19 patients. The primary tumor was staged pT1 in 2 patients, pT2 in 29, pT3 in 19, and pT4 in 16. The overall incidence of lymph node metastases was 79% (9 pN1, 3 pN2a, 33 pN2b, 7 pN2c) which was not correlated with T stage (50% pT1, 72% pT2, 89% pT3, 81% pT4). Occult nodal metastases were present in 42% of cases (8/19) with an incidence that increased from 11% (1/9) for pT1-2 to 70% for pT3-4 (7/10). The bilateral metastases (11%) were uniformly distributed between strictly lateral neoplasms and those tumors involving the midline. The incidence of bilateral metastases reached 19% only in patients with T4 cancers. Occult controlateral metastases were found in 12% of patients not having clinical evidence of metastases on the contro-lateral side of neck dissection (4/33). Nodal metastases never involved the I and V levels. Our data did not permit an assessment of the incidence of retropharyngeal lymph node metastases. In view of these results and considering current knowledge of the anatomy of lymphatic drainage, a selective II-IV dissection extending to the level VI on the side of the tumor appears justified in cases clinically staged as NO. In our view, when the lesion involves the posterior wall of the pharynx, neck dissection should be extended to the lateral retropharyngeal lymph nodes. Selective dissection of the controlateral side of the neck should be performed in patients having either locally advanced primary lesions or with lesions approaching the midline. In the presence of metastases which are either clinically or intraoperatively evident, neck dissection should be extended to additional lymph node levels. PMID- 11938707 TI - [Role of gastroesophageal reflux in precancerous conditions and in squamous cell carcinoma of the larynx: our experience]. AB - The major risk factors for the onset of precancerous lesions and squamous cell carcinoma of the larynx are, above all, tobacco smoke, alcohol abuse and exposure to viral and toxic agents. In recent years, however, gastro-esophageal reflux (GER) has also aroused significant interest not only as carcinogen but also as co carcinogen in association with smoking and alcohol consumption. The purpose of the present work is to provide an objective evaluation of the presence of distal and proximal esophageal reflux using multi-electrode pH monitoring in patients with precancerous lesions of the larynx and laryngeal and pharyngolaryngeal neoplasms. A total of 24 patients consecutively hospitalized during 2000 were evaluated: 20 with squamous cell carcinoma of the larynx and/or pharynx-larynx and 4 with precancerous vocal cord lesions. All the patients provided a case history using a validated questionnaire and underwent electronic videolaryngoscopy and 24-hour pH monitoring. Data analysis showed that in 83.3% of the cases (20/24) pH monitoring was indicative of pathological GER and 63.7% of these patients had no complaints related to reflux. Moreover, 7/24 patients have had a previous gastrectomy (Billroth II) strictly related to the presence of bile reflux. The Authors conclude that, according with recent literature, pH monitoring data further indicate that patients with precancerous or neoplastic pharyngeal-laryngeal lesions present often a positive history of GER, even though no direct cause-effect relationship can yet be established because of the co existence of other important risk factors such as smoking and alcohol consumption. Additionally, the achlorhydria found in 7/24 patients with an history of gastrectomy could suggest a possible, as yet unproven, role of other reflux components--in particular, of bile (alkaline) and chlorhydro-peptic (acid) components--as a risk factor or co-factor for tonset of precancerous and/or neoplastic lesions of the larynx. PMID- 11938708 TI - [Bilobed flap in the reconstruction of cheek defects]. AB - The bilobed flap is a reliable means for the reconstruction of facial defects in one stage. This reconstruction technique has been applied on a 69-years-old man affected by carcinoma of the cheek, involving overhanging skin and underlying oral mucosa. Stensen's duct reconstruction was also performed with a vein graft and, 8 months later, deviation of the rima oris was corrected by inserting a goretex strip at the level of the upper lip elevator muscle. The present work considers the bilobed flap as a means for cheek defects reconstruction compared with other reconstruction techniques. PMID- 11938709 TI - [Malignant fibrous histiocytoma of the larynx. Presentation of a clinical case and review of the literature]. AB - Malignant Fibrous Histiocytoma (MFH) is a tumor of mesenchymal origin which rarely occurs in head and neck. At the laryngeal level this neoplasm manifests itself as a sessile or peduncolate mass with ulcerations. Histological diagnosis of small biopsy samples is difficult and is, therefore, performed on the full surgical piece. The treatment of choice is complete exeresis of the mass within a safety margins. The Authors present a case report, the 37th laryngeal MFH reported in the literature and, in particular, the 5th case presented in a supraglottic site. Surgical treatment performed in this patient was exeresis of the neoplasm through pharyngothyrotomy conduced with safety margins and reconstruction by thyrohyoidpexy. PMID- 11938710 TI - [Primary tuberculosis of the middle ear: description of 2 cases and review of the literature]. AB - Recently, a recrudescence of tuberculosis (TBC) as been found, even in Western world and in Italy, most likely in relation to the increase in immigration from developing countries and to the pathologies that cause immunodepression. The present paper reports two cases of primitive TBC of the middle ear, both coming under observation for facial paralysis. These cases highlight the clinical therapeutic features of this disease, the difficulty in diagnosis and the need to include TBC in the differential diagnosis of phlogistic processes of the middle ear. The clinical-diagnostic features of presented cases focus attention on the consideration of TBC in the differential diagnosis of chronic otitis, particularly in presence of unusual symptoms, such as facial paralysis, and in case of otitis with rapid onset, unresponsive to conventional treatment. In one case, first examination of the ear showed normal tympanic membrane (absence of any tympanic perforation) and normal timpanometry; these facts steered diagnosis away from a middle ear phlogosis pathology. Both cases showed how radiological semeiology, CT and NMR are not enough to diagnose the nature of the disorder, particularly in case of cholesteatoma. Moreover, both patients were in good general conditions at the time of the examination, they didn't show history of previous immunodepression or other risk factors. The cases reported call attention to the aspecific characteristics of the onset of primitive tuberculous otitis media and difficulty in making a diagnosis. They underline the need for post-operative anti-tuberculous chemotherapy which provided a rapid and definitive resolution of infectious process. PMID- 11938712 TI - Coronal dentinal tubules of non-erupted deciduous incisors. AB - The diameter and density of coronal dentinal tubules were assessed in non-erupted human deciduous incisors. Using liquid nitrogen, frozen samples were fractured in longitudinal and oblique directions and observed by means of a Jeol JSM-6100 scanning electron microscope. It was observed that the diameter of dentinal tubules ranged from 0.50 to 1.94 micrometers and increased significantly from the outer to the inner layers. However, comparing the incisal, middle and cervical thirds, no statistically significant differences were observed. The number of tubules per square millimeter varied from 3,281 to 32,313 and also increased from the external dentinal surface to the dentinal surface of the pulp chamber. PMID- 11938713 TI - [Oral manifestations, CD4+ T-lymphocytes count and viral load in Brazilian and North-American HIV-infected children]. AB - Oral manifestations related to HIV infection were evaluated in 184 children of both genders, aging 0-13 years. The oral exams were carried out at the Ambulatory of Pediatric AIDS, Sao Lucas Hospital and at the Clinics Hospital, both in Porto Alegre, RS, Brazil, and at the University Hospital and School of Dentistry, State University of New York, Stony Brook, NY, USA, from January 1999 to May 2000. A high prevalence of oral manifestations was observed both in Brazilian (72.73%) and American (53.66%) children. The most frequent manifestations were cervical lymphadenopathy, swelling of the parotids, erythematous candidiasis, petechia, gingivitis, xerostomia and angular cheilitis. Children with oral manifestations had almost normal counts of CD4+ T lymphocytes and high viral load. PMID- 11938714 TI - Biometric study of the width, length and depth of the root trunk groove of human lower second molars. AB - One hundred human lower second molars, 51 from the left side and 49 from the right side, extracted due to their poor clinical and radiographic conditions were utilized in this study. Using a Contracer apparatus, the profiles of the buccal and lingual root surfaces of these teeth were traced on a millimeter-scaled paper. The profiles were registered from the cementoenamel junction (CEJ), millimeter by millimeter, up to the entrance of the furcation. The width, length and depth of the root trunk groove, as well as the length of the root trunk, were studied. After statistical analysis (p < 0.05) it was possible to conclude that: a) the mean width of the root trunk groove on the buccal surface was 3.6 mm and, on the lingual surface, 3.3 mm; b) the mean depth of the root trunk groove on the buccal surface was 0.88 mm and, on the lingual surface, 0.77 mm; c) the mean length of the root trunk groove on the buccal surface was 2.93 mm and, on the lingual surface, 3.61 mm. The mean length of the root trunk on the buccal surface was 3.09 mm and, on the lingual surface, 3.91 mm (p < 0.025). There was a coincidence between the length of the root trunk and that of its groove in 90.2% of the buccal surfaces of the samples from the left side, and in 77.5% of the samples from the right side; on the lingual surface, the coincidence occurred in 77.5% of the teeth from the left side, and on 88.3% of the teeth from the right side. This work revealed that there is a concavity on the root trunk region of the lower second molar, whose depth and width were greater on the buccal surface and whose length was greater on the lingual surface. The depth of the root trunk groove increased in the apical direction, with maximum depth in the last millimeter of the root trunk. The root trunk was longer on the lingual surface than it was on the buccal surface. PMID- 11938715 TI - In vitro effect of intracanal medicaments on strict anaerobes by means of the broth dilution method. AB - The determination of bacterial susceptibility to intracanal medicaments is a necessity. Nevertheless, few studies utilize the proper methodology to carry out that evaluation with anaerobes. In this study, the steps of a broth dilution method, carried out in microplates (microdilution) and tubes (macrodilution), to test the effect of traditional intracanal medicaments on anaerobic bacteria are described. The results are presented as values of minimal inhibitory and bactericidal concentrations (MIC and MBC). Standardized inocula of the anaerobic bacteria Prevotella nigrescens (ATCC 33563), Fusobacterium nucleatum (ATCC 25586) and Clostridium perfringens (ATCC 13124), in reinforced Clostridium medium (RCM) and supplemented Brucella broth, were submitted to different concentrations of calcium hydroxide, chlorhexidine digluconate, camphorated paramonochlorophenol and formocresol solutions. The drugs were diluted in the same culture broths, in microplates and tubes, and were then incubated in anaerobiosis jars at 37 degrees C for 48 or 96 hours. The determination of MICs was carried out through visual and spectrophotometric readings, and the determination of MBCs, through the plating of aliquots on RCM-blood agar. For that kind of study, the macromethod with spectrophotometric reading should be the natural choice. MICs and MBCs obtained with the macromethod were compatible with the known clinical performance of the studied medications, and the values varied according to the bacteria and culture media employed. RCM was the most effective medium and C. perfringens, the most resistant microorganism. PMID- 11938716 TI - Tensile bond strength of adhesive systems--effects of primer and thermocycling. AB - The objective of this study was to evaluate the effects of primer and thermocycling on the bond strength of multi-purpose adhesive systems applied to enamel, under tensile stress. The following bonding systems were applied, according to the manufacturers' instructions, on unground enamel buccal surfaces of 96 premolars, with or without the application of primer: Scotchbond MP, OptiBond FL, Amalgambond Plus and OptiBond (dual-cure). A composite resin (Z100, 3M) was applied and light-cured in a cast metal hollow cone, which was previously fixed to the enamel surfaces. Half of the sample was subjected to 3,000 thermocycles (5-37 degrees C; 37-55 degrees C, dwell time of 60 s), and the other half was stored in water at 37 degrees C for the same period. The data were treated by means of ANOVA and no significant effects were detected, which indicates that tensile bond strength was not affected by the adhesive systems, application of primer or thermocycling. PMID- 11938717 TI - [Influence of post-fixation time on shear bond strength of brackets fixed with different materials]. AB - The purpose of this study was to evaluate the shear bond strength of four bonding materials for brackets, 10 minutes and 24 hours after their fixation, as well as the kinds of fracture observed. The buccal surfaces of 32 premolars were etched for 30 seconds with 35% phosphoric acid, and brackets were bonded using Orthodontic Concise (3M) and Z100 (3M). In other 32 premolars, brackets were bonded with Fuji I (GC) and Fuji Ortho LC (GC) on the buccal surfaces, without acid etching. After the bonding procedures, 32 samples were stored in distilled water at 37 degrees C for 10 minutes, and 32 samples were stored at the same temperature for 24 hours. The samples were then submitted to shear bond strength testing in an Instron testing machine, at a crosshead speed of 0.5 mm/min. The results were submitted to ANOVA and Tukey's test (p < 0.05). The results indicated that, for 10 minutes and 24 hours, Orthodontic Concise showed shear bond strength values (6.22 and 7.73 MPa, respectively) higher than Fuji Ortho LC (3.32 and 5.10 MPa), Z100 (2.72 and 4.51 MPa) and Fuji I (2.52 and 4.54 MPa). No statistical difference was verified between Fuji Ortho LC, Z100 and Fuji I (p < 0.05). In conclusion, Orthodontic Concise showed better shear bond strength values than the other three bonding materials, for both 10-minute and 24-hour storage periods; the shear bond strength averages were higher for the storage time of 24 hours, for all tested materials, and a great number of adhesive fractures were observed in the specimens which received Fuji I, Orthodontic Concise and Z100. PMID- 11938718 TI - [Prevalence of root caries and periodontal conditions in an elderly institutionalized population from Piracicaba-SP]. AB - Preventive and invasive dental procedures have improved in the last two decades, leading to a greater longevity of teeth. As a result, teeth are more susceptible to periodontal diseases and, consequently, to root caries. The purpose of this study was to verify the prevalence of both periodontal diseases and root caries in an institutionalized population from Piracicaba-SP, Brazil. In this study, 151 subjects, aging 54 to 93 years, were examined. The sample was divided into two groups: a group of subjects aging 50 to 75 years and a group of subjects over 75 years old. The sextants were examined and classified as to periodontal health, periodontal treatment needs, prevalence of gingival recession and root lesions. In conclusion, a high prevalence of root caries was verified. That indicates an increase in the susceptibility to these lesions, which is associated with the aging process. PMID- 11938719 TI - [Self-efficacy as a mediator for improvement in oral health clinical indices]. AB - The concept of self-efficacy (SE) was developed by BANDURA, in 1977. SE has been widely utilized in health care and has shown to be a powerful predictor in various domains of behavior such as in smoking cessation, weight control and participation in programs of disease prevention. This study investigated if a psychosocial intervention fosters improvements in clinical indices in patients who are about to receive dental treatment. It was predicted that the experimental group (EG) would show a greater progress as to plaque and bleeding clinical indices than the control group (CG), and SE would mediate that improvement. Besides the standardized dental procedures, the 30 participants from the EG received a psychosocial intervention based on the PROCHASKA; DiCLEMENTE'S model (1983). The 30 participants in the CG received only standardized dental procedures. The results revealed that the prevalence of scores 2 and 3 of the plaque and bleeding indices underwent a significant decrease in the EG, in relation to the CG (U = 97.00, p = 0.0001 and U = 179.00, p = 0.0001, respectively). Only the subjects from the EG presented a relevant increase of SE, considering the pre- and post-treatment periods (Z = -3.58, p < 0.0001). Yet, there was no difference between both groups as to the increase of SE between the pre- and post-testing periods. In short, the results showed that psychosocial intervention was effective and suggest that other factors besides SE may mediate a relevant improvement of clinical indices in the EG. PMID- 11938720 TI - [Analysis of the point-counting and planimetric methods in the quantification of denture biofilm--a methodological validation study]. AB - Two methods of quantification of the biofilm (point-counting and planimetric) were tested and compared with the paper-weighing method and with the Prosthesis Hygiene Index. The internal surfaces of 62 complete dentures were stained and photographed. The slides were projected on a paper sheet. The total area and the area covered with biofilm were contoured using a black pencil. The point-counting method (experimental 1) was carried out on a mesh of equidistant points. For the planimetric method (experimental 2), the areas of interest were measured by means of a digital planimeter. In the paper-weighing method (control 1) the areas of interest were cut and weighted on a precision scale. In the determination of the Prosthesis Hygiene Index (control 2), the accumulation of biofilm was estimated by means of a scoring method. The results revealed an agreement rate of 82% between the paper-weighing method and the point-counting method, and an agreement rate of 95% between the former and the planimetric method, which was confirmed by high coefficients of correlation (0.98 and 0.99, respectively). The comparison with the Prosthesis Hygiene Index resulted in 55% of agreement with the point counting method and in 37% of agreement with the planimetric method. The experimental methods can be useful in clinical studies involving the evaluation of the performance of denture cleansers. PMID- 11938721 TI - [Osteosarcoma and chondrosarcoma--radiographic differentiation with computerized tomography]. AB - The purpose of this paper was to associate and compare the radiographic patterns of osteosarcoma and those of chondrosarcoma by means of computed tomography (CT). Computed tomographs of five cases of osteosarcoma and five cases of chondrosarcoma of the head and neck region were analyzed. The sensitivity of CT was calculated based on the results of histopathological examinations. We concluded that CT can facilitate the establishment of the final diagnosis since it is possible to distinguish the individual characteristics of osteosarcomas and chondrosarcomas in that exam. PMID- 11938722 TI - Oral manifestations in children with AIDS and in controls. AB - Thirty children with AIDS, aging 2 to 6 years, of both genders, treated as outpatients at the hospital Santa Casa de Misericordia, Sao Paulo, were evaluated for oral manifestations and compared to a control group of age- and sex-matched healthy subjects. The correlation between oral lesions and the degree of immunosuppression was analyzed. The most prevalent oral manifestations- lymphadenopathy, followed by gingival alterations--were observed in the children with the highest levels of immunosuppression. Other manifestations observed in the AIDS group were: pseudomembranous and erythematous candidosis, enlargement of the parotids and ulcers. The results did not indicate a higher frequency of lesions. However, the observed oral manifestations had an early occurrence, which indicates that the early diagnosis is an important component in the management of those patients. PMID- 11938723 TI - [Study of the aluminum-zinc alloy as an alternative method of filtration in dental X-ray devices]. AB - The aim of this study was to evaluate the effects of alternative filtration with the aluminum-zinc alloy in dental X-ray devices, regarding the energy spectrum, dose, contrast and image quality. This study was carried out comparing the conventional aluminum filter with aluminum-zinc alloy filters of various thicknesses and with different percentages of zinc in the alloy (2, 3, 4 and 5%). A conventional X-ray device operating with 60 and 70 kVp as well as D- and E speed films were utilized in order to obtain images of a phantom and an aluminum step wedge. The utilization of Al-Zn filters resulted in a spectrum without hardening of beams. The rate of kerma in the air decreased 18.53%, without any alteration in contrast and image quality. The results revealed that the 2% Al-Zn alloy filter with 1.99 mm of thickness, utilized with the tension of 60 kVp and D speed film, had the best performance. It reduced the rate of kerma without varying the indices of contrast and maintained image quality at acceptable levels, without increasing the time of exposure. PMID- 11938724 TI - [Comparative study of microleakage in class V restorations]. AB - The aim of this study was to analyze the laboratorial performance of dentin bonding agents, glass ionomer cements and compomers, regarding the sealing of the gingival margins of class V cavities, prepared on the cementoenamel junction. Standardized class V cavities (4 x 3 x 2 mm) were prepared on the buccal and lingual/palatal surfaces of 20 extracted human third molars. Forty cavities were restored with different combinations of materials, as follows: group 1 (G1)- Vitremer (3M); group 2 (G2)--Vitremer (3M) and Syntac Sprint/Tetric Ceram (Vivadent); Group 3 (G3)--Syntac Sprint/Tetric Ceram (Vivadent); Group 4 (G4)- Prime & Bond 2.1/Variglass (Dentsply). After thermocycling in methylene blue dye, the specimens were longitudinally sectioned and analyzed with a stereoscopic lens in order to evaluate the leakage on the gingival margin. The obtained data were analyzed with the Mann-Whitney test, and the results revealed significantly better sealing (p = 0.05) in G1, when compared with G4. No significant differences were seen between the other groups. Under the given conditions, Vitremer offered better sealing of the gingival margin than the system Prime & Bond 2.1/Variglass. PMID- 11938725 TI - [Comparison of 2 techniques for enamel remineralization]. AB - Six patients enrolled at the Hospital of Rehabilitation of Craniofacial Anomalies, University of Sao Paulo, aging 16 to 20 years, were selected. All patients presented with white spots in both sides of the superior arch, which totalized 41 spots. The left side of the arch was submitted to remineralization by means of the conventional technique with Reminer, a commercially available product. In the right side, a modified technique was employed, with the utilization of fluoridated dentifrices every night, during 8 weeks. It was observed that successful remineralization occurred in 62% of the spots when the conventional technique was employed, whereas, when fluoridated dentifrices were utilized, there was partial remineralization in 10% of the spots. The utilization of Reminer was more effective than the utilization of fluoridated dentifrices. PMID- 11938726 TI - [Construction of three transgenic vectors carrying the latent membrane protein gene of Epstein-Barr virus]. AB - In order to produce transgenic mice carrying the latent membrane protein(LMP) gene of Epstein-Barr virus(EBV) and to study the oncogenic role of LMP gene in vivo, three different transgenic vectors carrying the LMP gene were constructed. In pBR322-MT-LMP vector, the promoter of LMP gene is the regulation region of mouse metallothionein-I gene. In pMci3-LMP shuttle vector, the promoter of LMP gene is the replication origin(oriP) of EBV. In pMV-LMP-c-myc retroviral vector, the long terminal repeat (LTR) of mouse sarcomavirus serves as the promoter of LMP gene. The characterization and usefulness of these three transgenic vectors are also discussed. PMID- 11938728 TI - [Determination of vitamin E in juices by high performance liquid chromatography (HPLC)]. AB - A modified method for the determination of tocopherol(vitamin E) in juices by high performance liquid chromatography(HPLC) has been proposed in this paper. The juice samples were deproteinized with ethanol and extracted with hexane. Organic layer was evaporated to dryness with nitrogen after it was separated. Then the residue was redissolved with methanol and injected into a reversed-phase HPLC equipped with a C18 column. The tocopherols were eluted with methanol mobile phase at a flow rate of 1.0 ml.min-1 and detected by fluorescence(lambda exc = 295 nm, lambda em = 330 nm). A good linearity was shown between the concentration of tocopherol and the peak height at the concentration range of 0.2-2.2 micrograms.ml-1(r = 0.9995, n = 5). The recovery rate was more than 96.8%, and impurities in the samples were separated completely. This method is sensitive, simple and stable. PMID- 11938727 TI - [Effects and applications of ascorbic acid on the proliferation of Chlamydia trachomatis]. AB - Two hundred and eighty-eight samples of patients with nongonococcal urethritis (NGU) and prostatitis were detected by cell culture and immunofluorescence assay for Chlamydia trachomatis (CT) and the effects of ascorbic acid(vitC) on the formation rate of inclusion of CT in positive samples were also studied. The results showed that the formation rate of inclusion of CT was 29.5% when the concentration of vitC was 5 micrograms.ml-1. The difference between test group and control group which contained cycloheximide in the media was insignificant (P > 0.05). The results suggested that vitC was a kind of nutrient needed for CT. The proliferation of CT in the cell can be promoted by vitC. We can replace cycloheximide by vitC in McCoy cell culture to detect clinic samples with CT. PMID- 11938729 TI - [Molecular typing of staphylococcus aureus with randomly amplified polymorphic DNA]. AB - The randomly amplified polymorphic DNA(RAPD) was used to type the 26 clinically isolated strains of staphylococcus aureus. The results were that all three primers of this method, ERIC-2, Ap1, and Ap7 on molecular typing of staphylococcus aureus and identified 9, 6, and 10 genotypes, respectively; the primer Ap7 produced better results than the other primers. The present study suggests that RAPD is a simple and rapid method and has a practical value in the molecular typing of staphylococcus aureus. PMID- 11938730 TI - [Relationship between smoking and drinking and herniation of intervertebral disc]. AB - To study whether there is relationship between smoking and drinking and herniation of intervertebral disc, we investigated 214 patients who had been proved with herniation of intervertebral disk during 1986-1996. The results were that in the younger (< or = 35 year old), no matter whether they drank or not, the number of smoker in the patients with herniation of intervertebral disc was more than that in the control group; no matter whether they smoked or not, the number of drinker in the patients with herniation of intervertebral disc was more than that in the control group. PMID- 11938731 TI - [Experimental study on induction of apoptosis of leukemic cells by Boswellia carterii Birdw extractive]. AB - The purpose of the study was to investigate the apoptosis of leukemic cells induced by Boswellia Carterii Birdw(BCB). The target leukemia cell line HL60 and bone marrow leukemic cells from 30 acute non-lymphocytic leukemic(ANLL) patients (3 M1 11 M2a 10 M3 1 M4a 5 M5b) were studied. Apoptosis was detected by morphological observation, DNA electrophoresis, percentage of DNA fragmentation test and flow cytometric cell cycle analysis. It is concluded that BCB can induce apoptosis in ANLL cells and HL60 cells. PMID- 11938732 TI - [Affection of aprotinin on tissue factor pathway inhibitor in peri-operation of cardiac direct vision operation]. AB - Twenty patients underwent cardiac operation with cardiopulmonary bypass(CPB). We collected their blood samples during peri-operation and measured tissue factor pathway inhibitor(TFPI) and active coagulation time(ACT). The results were that TFPI and ACT were significantly increased after heparin injection and during CPB. The changes of TFPI and ACT were similar and pathophysiological; it was evident that heparin stimulated TFPI release and ACT prolonged during cardiac operation with CPB. Aprotinin did not affect the change of TFPI. The results suggest that TFPI has the effect on anticoagulation during CPB. PMID- 11938733 TI - [Affect of insulin-like growth factor I and estradiol on the growth of uterine leiomyoma]. AB - To study roles of insulun-like growth factor 1(IGF-I) and estradiol(E2) in the growth of leiomyoma. Protein expressions of IGF-I receptor(IGF-I-R), estrogen receptor(ER), and cell proliferation associated nuclear antigen(ki-67) were analysed by streptavidin-peroxide method in 40 cases. Serum level and local concentration of IGF-I were determined by immunoradiometric assay(IRMA) in 20 cases. Serum level of estradiol was determined by radioimmunoassay in 20 cases. Twenty normal fertile women were studied as control. Protein expressions of IGF-I R and ER in uterine leiomyoma were significantly higher than in myometrium(P < 0.05); and local concentration of IGF-I positively correlated with ki-67 (r = 0.6513, P < 0.05); and expression of estrogen receptor. The serum levels of IGF-I and E2 had no significant difference between leiomyoma patients and normal women. IGF-I and E2 promote the growth of leiomyoma and IGF-I may act as a mediator of estrogen. PMID- 11938734 TI - [Studies on ACE gene insertion/deletion polymorphism, serum ACE activity, and diabetic retinopathy in type II diabetic patients]. AB - To clarify the relationship among angiotensin 1-converting enzyme(ACE) gene insertion/deletion(I/D) polymorphism, serum ACE activity, and diabetic retinopathy in Type II diabetic patients, we examined 36 healthy controls, 40 Type II diabetic patients without diabetic retinopathy, and 68 Type II diabetic patients with diabetic retinopathy. All of patients suffered from Type II diabetes beyond 5 years and matched in age and body mass index. An insertion/deletion polymorphism of ACE gene was identified by polymerase chain reaction(PCR). Serum ACE activity was determined using spectrophotometry. The distribution of DD, ID, and II genotypes of the ACE gene did not differ among the three groups (16.7%, 33.3%, and 50.0% in the healthy controls; 22.5%, 35.0%, and 42.5% in Type II diabetic patients patients without diabetic retinopathy; and 20.6%, 30.9%, and 48.5% in Type II diabetic patients with diabetic retinopathy; respectively). The frequency of ACE I/D genotypes was not significantly different between non-proliferative retinopathy and proliferative retinopathy of Type II diabetic patients with diabetic retinopathy (P > 0.05). Serum ACE activity of three groups was similar (P > 0.05). These results do not support the hypothesis that the DD genotype of the ACE gene would be a clinically useful genetic marker for predicting the development of diabetic retinopathy in Type II diabetic patients. There is no association between ACE gene I/D polymorphism and the prognosis of diabetic retinopathy in Type II diabetic patients. ACE may not involve in forming diabetic retinopathy in Type II diabetic patients. PMID- 11938735 TI - [Alteration of blood glucose, insulin and lipid in the offsprings of patients with essential hypertension]. AB - We prospectively investigated the serum insulin, glucose and serum lipoproteins in 40 normotensive offsprings(test group) of parents with essential hypertension compared with 35 normotensive offsprings (control group) of the normotensive families. Compared with control group, the test group was significantly elevated in fasting serum insulin levels(6.82 +/- 2 vs 4.02 +/- 1.7 microU.ml-1) (P < 0.05), fasting serum glucose(4.32 +/- 0.4 vs 3.84 +/- 0.5 mmol.L-1), serum total triglycerides(1.97 +/- 0.8 vs 1.32 +/- 0.5 mmol.L-1), total cholesterol(Tch) (5.13 +/- 1.0 vs 4.24 +/- 0.6 mmol.L-1), low density lipoprotein cholesterol (2.82 +/- 1.0 vs 2.24 +/- 0.7 mmol.L-1), very low density lipoprotein cholesterol (1.02 +/- 0.4 vs 0.66 +/- 0.27 mmol.L-1), high density lipoprotein cholesterol/Tch ratio (0.27 +/- 0.08 vs 0.33 +/- 0.07 mmol.L-1). While post glucose load the serum insulin and glucose level after glucose load were much higher (P < 0.01) in the test group than that in controls. The findings demonstrated that young normotensive offsprings of parents with hypertension had excellent health, they had an impairment of insulin-mediated glucose disposal, hyperinsulinemia, and dyslipidemia. These findings supported that a common hereditary defect exits in familial trait for essential hypertension and disturbance of carbohydrate and lipoprotein metabolism may be detected before or at least at very early stage of the development of essential hypertension. PMID- 11938736 TI - [Changes of serum cardiac troponin T in patients undergoing cardiac surgery]. AB - We detected the changes of serum cardiac troponin T(cTn T) content and serum cardiac enzyme activities in 15 patients with cardiopulmonary bypass during cardiac operation. The central venous blood samples were collected to measure the serum concentration of cTn T by enzyme immunoassay methods and activities of lactate dehydrogenase(LDH), creatine kinase(CK), and creatine kinase isoenzyme(CK MB) by biochemical methods. The content of cTn T and activities of LDH, CK, and CK-MB increased significantly after cardiac surgery. There was positive correlation between cTn T and aortic cross clamping time (r = 0.6014, P < 0.05), but there was no correlation between CK-MB and aortic cross clamping time (r = 0.4712, P > 0.05). We conclude that serum cTn T and CK-MB can monitor cardiac ischemic reperfusion injury and can evaluate myocardial protection methods during cardiac operation, but cTn T is better than CK-MB. PMID- 11938737 TI - [Significance of detection of HBV-DNA and CMV-DNA by polymerase chain reaction in screening mothers' milk]. AB - Hepatitis B virus(HBV) DNA and cytomegalovirus DNA in the colostrum of lying-in women were detected by polymerase chain reaction for the safety of mothers' milk in the milk bank. The results showed that: 1. HBV-DNA was not found in the milk of women with negative serological markers of the virus, while in the milk of women with positive marker of HBsAg, HBeAg and HBcAb, the positive rate of HBV DNA was 81.25%, in the milk of mothers with positive marker of HBsAg, HBcAb and HBeAb, the positive rate was 45.24%, in the milk of women only with HBcAb and HBeAb positive, the rate was 23.08%. 2. The specific amplified products of CMV DNA in the milk were obtained in 42.80%-70.00% women whose CMV IgM antibody was positive. These findings suggest that the excretion of viruses would increase in mothers with active virus replication, and the viruses would be transmitted to newborns by milk. The authors think that only the milk from the mothers with both negative serological markers and negative PCR of the viruses can be collected and stored in the milk bank. PCR is a specific and sensitive technique which can be used to monitor the safety of mothers' milk. In this paper, the authors have presented some suggestions about how to give breast-feeding and protect newborns whose mothers are various status of virus secretion in the milk. PMID- 11938739 TI - [Effect of hydrogen peroxide on expression of interleukin-2 gene in mouse splenocytes]. AB - The effect of hydrogen peroxide on interleukin-2(IL-2) production and IL-2 mRNA expression in mouse splenocytes was detected by cellular immunological and molecular biological methods. The results were that the level of IL-2 activity was significantly elevated in 10(-4) mol.L-1 of H2O2 as compared with the control group (P < 0.01), was obviously inhibited in 10(-3)-10(-2) mol.L-1 of H2O2(P < 0.01), and had no effect in 10(-8)-10(-5) mol.L-1 of H2O2. The results of dot blotting were that 10(-4) mol.L-1 of H2O2 enhanced IL-2 mRNA expression, while 10(-3) mol.L-1 inhibited it, and 10(-5) mol.L-1 had no effect in mouse splenocytes. These results suggest that the effect of H2O2 on IL-2 production may be mediated by regulating IL-2 mRNA expression in mouse splenocytes. PMID- 11938738 TI - [Efficacy of TACE in treatment of intraportal tumor thrombi with double helical CT]. AB - The present study observed alterations of the intraportal tumor thrombi before and after transcatheter hepatic arterial chemo-embolization(TACE) in 36 patients with massive hepatocellular carcinoma(HCC). Elscint double helical CT was used to scan liver in the portal phase before and after TACE. The results showed that the scan technique could be used to evaluate the distribution of intraportal tumor thrombi. After treating with TACE, retention of iodized oil was found in the intraportal tumor thrombi(14/36), and the intraportal tumor thrombi were reduced in 28 out of 36 cases. The survival rate was more than 1 year in 20 patients. These data indicate that the blood supply of the intraportal tumor thrombi comes from hepatic arterial system. TACE seems to be valuable for the treatment of intraportal tumor thrombi with massive HCC. PMID- 11938740 TI - [Use of color Doppler esophageal ultrasonography for assessing hemodynamics of the lower esophageal veins before and after endoscopic esophageal variceal ligation]. AB - Using color Doppler esophageal ultrasonography(CDEUS) with water balloon, we examined the lower esophageal veins of 24 patients with portal hypertension and esophageal varices. The results showed that CDEUS with water balloon could clearly display the lower esophageal veins and could effectively evaluate the changes of hemodynamics of the lower esophageal veins before and after endoscopic esophageal variceal ligation. The maximum blood velocity of peri-esophageal vein in the active bleeding group was higher than that in the non-active bleeding group. PMID- 11938741 TI - [Effect of diazepam on fetus after intravenous administration during the active phase in labour]. AB - To investigate the effects of diazepam on fetal heart rate and Apgar scoring administered intravenously during the active phase in labour, sixty-two normal term parturients during the active phase of labour with the cervical dilation to 3-5 cm were randomly divided into two groups, diazepam group (n = 32) and control group (n = 30). In diazepam group, 10 mg diazepam was administered intravenously. The concentrations of diazepam in maternal and cord serum were measured by high performance liquid chromatography after birth and the fetal heart rates were monitored by electronic monitor before or after administration of diazepam and Apgar scores were determined immediately after birth. The results showed that the mean level of diazepam in umbilical cord serum was markedly higher than that of maternal serum. Fetal heart base line mutation in diazepam group was weaker than that of the control group. Apgar scores in the neonates of diazepam group was lower than those of the control group. The results indicate that the use of diazepam in pregnant women during active phase of labour may cause depression of fetus. PMID- 11938742 TI - [The role of the leukocytes in pathogenesis of secondary brain injury]. AB - The levels of leukocyte count, neutrophil number, leukocyte adhesiveness, leukocyte releasing oxygen-free radical (O2.-) after phagocytosing bacteria and serum lipid peroxide MDA were determined in the peripheral blood of 40 patients with acute head injury within 48 hours and 22 normal adults. These levels of patients were significantly higher than those of the controls (P < 0.05), and increased with the severity of craniocerebral injury elevation. The leukocyte count and neutrophil number of those patients who died were significantly higher than these of those survived patients (P < 0.05). It suggested that leukocytes aggravated the secondary brain injury after craniocerebral injury by increasing of leukocyte adhesiveness, and generating and releasing oxygen-free radical, and measurement of leukocyte count and neutrophil number in the peripheral blood could evaluate the prognosis of the patient with craniocerebral injury. PMID- 11938743 TI - [Preservation of hearing and facial nerve function after Gamma knife therapy for acoustic neurinomas]. AB - Thirty-five patients with acoustic neurinomas were treated by Gamma knife from Sept. 1995 to Oct. 1997. Neuroimaging studies performed in 21 patients with more than 12 months showed that tumor size decreased in 9(43%) cases, unchanged in 11(51%) cases, and increased in 1(15%) case. The tumor growth control and regression rates were 95% and 43%, respectively. Some hearing was preserved in four patients (36%) who had hearing prior to Gamma knife treatment. There was new transient facial weakness in five patients(23%), and there was new trigeminal neuropathy after treatment in one patient (4.8%). These findings indicate that Gamma knife therapy is a safe and effective management for acoustic neurinomas, especially in preventing facial function and hearing. PMID- 11938744 TI - [The relationship between anticardiolipin antibodies and fetal intrauterine growth retardation]. AB - To investigate the relationship between anticardiolipin antibodies(ACA) and fetal intrauterine growth retardation(IUGR), serum ACA was determined in 60 pregnant women by enzyme linked immunosorbent assay(ELISA). Binding index(BI) of ACA-IgM or ACA-IgG in cases with IUGR(n = 30) was significantly higher than that in normal pregnant women(n = 30) (P < 0.05). There was no obvious relationship between the weight of neonate and BI of ACA-IgM(r = 0.1904), ACA-IgA(r = 0.1829) or ACA-IgG(r = 0.2013) in IUGR group. Increased ACA-IgM had a negative association with C3 (r = -0.4108, P < 0.05) and C4(r = -0.4092, P < 0.05), respectively. After routine therapy for IUGR during 27-35 weeks' gestation, the weights of neonates were 2436.87 +/- 437.29 gram in ACA-IgG positive group(n = 15) and 2905.33 +/- 195.95 gram in ACA-IgG negative group(n = 15). There was a statistical significance between these two groups(P < 0.01). It is concluded that ACA may play an important role in the pathogenesis of IUGR. The determination of ACA is also useful for early diagnosis and treatment of IUGR. PMID- 11938745 TI - [Relationship of age, sex, body weight, smoking, blood lipids and fibrinolytic activities in healthy subjects]. AB - Relationship of plasma activities of tissue-type plasminogen activator (tPA) and plasminogen activator inhibitor-1(PAI-1) with age, sex, body weight, smoking, blood lipids were studied in 131 healthy subjects. The results demonstrated that plasma PAI-1 activity increased with age and body weight. PAI-1/t-PA ratio was higher in smoking than in non-smoking subjects. With multivariate analyses, plasma PAI-1 activity was positively correlated with age, body weight index(BMI) in males, and positively correlated with low-density-lipoprotein-cholesterol(LDL) in females, and negatively correlated with plasma tPA activity. Plasma triglyceride, high-density-lipoprotein-cholesterol did not show a correlation to PAI-1 or tPA activity. It is indicated that age, BMI, blood lipids may contribute to moderate fibrinolytic activities and that smoking may affect fibrinolytic function. PMID- 11938746 TI - [Changes of erythrocyte membrane ATPase activities and plasma lipids in patients with coronary heart disease]. AB - Fasting plasma triglyceride (TG), total cholesterol(TC), low density lipoprotein cholesterol(LDL-C), high density lipoprotein-cholesterol(HDL-C), activities of erythrocyte membrane Na(+)-K(+)-ATPase, Ca(2+)-ATPase, Mg(2+)-ATPase, and intraerythrocytic calcium concentration were examined in 21 patients with coronary heart disease(CHD) and 21 normal controls. It was found that the plasma contents of TG, TC, LDL-C and intraerythrocytic [Ca2+] in patients with CHD were much higher while plasma contents of HDL-C, activities of erythrocyte membrane Na(+)-K(+)- and Ca(2+)-ATPase were much lower than those of normal controls. Mg(2+)-ATPase activity showed no significant difference between these two groups. The activities of erythrocyte membrane Na(+)-K(+)- and Ca(2+)-ATPase were negatively correlated with plasma TG, TC, LDL-C levels, respectively, and positively correlated with plasma HDL-C. The possible pathogenic mechanism is discussed on the basis of our results. PMID- 11938747 TI - [Intermittent-pulse treatment of onychomycosis with orally administered itraconazole]. AB - Fifty-four patients received an intermittent regimen of itraconazole (26 of them were finger onychomycosis and 28 toe onychomycosis) and were followed-up for nine months. The result showed that the clinical cure rate of fingernails was 88.5% and the mycologic cure rate was 96.4%, while the clinical cure rate of toes was 82.1% and the mycologic cure rate was 96.4%. Few adverse events were observed by 7.4%, just like nausea, gastric discomfort etc. It suggests that the pulse administration system is highly effective and has a few side-effects. PMID- 11938748 TI - [Technique of enzyme digestion adding brushing for isolating bronchial epithelial cells]. AB - This study improved the previous techniques of harvesting rabbit bronchial epithelial cells. With 0.05% of trypsin on the epithelial side of trachea and bronchus, a mild digestion was used before a brushing protocol. The cells were identified with immunocytochemistry and electron microscope, both the cell viability and the membrane integrity of cells were evaluated. A comparison analysis of this method with the simple mechanic brushing and the simple digestion method were done. The results showed that, with this method, cells in a high purity and high viability could be obtained and the recovered cell number could be enough for experiments. It can be concluded that the method is useful and effective for isolating bronchial epithelial cells especially for small animals. PMID- 11938749 TI - [The model of piggyback orthotopic liver transplantation in pigs]. AB - To establish the pig model of piggyback orthotopic liver transplantation(PBOLT), twelve cases of pig PBOLT were performed. The result showed that ten pigs were survived after operation, and the operative successful rate was 83%. The mean operation time and mean anhepatic-phase time were 7 hours and 34 minutes, respectively. The mean bleeding amount was 1200 ml. The appearance and tissue structure of liver in every stage under optic microscope were normal. The results indicate that the pig model of PBOLT is feasible and that it is important to decrease the anhepatic-phase time for a successful liver transplantation. PMID- 11938750 TI - [Study on etiologic agents of vaginitis]. AB - In order to provide scientific basis for preventing and curing vaginitis, etiologic agents were examined in 354 patients with vaginitis and 115 healthy women using rapid and reliable laboratory methods. The inducements of vaginitis were analysed. The results showed that candida was only detected for the normal control group. Bacteria, candida and trichomonas were the commonest causes of vaginitis. With the increasing ages of patients with vaginitis, the incidence rates of all kinds of vaginitis were lowered. Vaginitis is associated with pregnancy, using antibiotics and contraceptive, or sexual activity. PMID- 11938751 TI - [Treatment of traumatic carotid-cavernous fistula using detachable balloon embolization]. PMID- 11938752 TI - [Pathogenesis and re-operation for residual defect after repair of ventricular septal defect]. PMID- 11938753 TI - [An approach to treat child intermittent exotropia with extraordinary recession of the external rectus]. PMID- 11938754 TI - [Effects of high-intensity periodic training on activity of serum phosphoglucose isomerase and hemorheology]. PMID- 11938755 TI - [Modulation of endothelin-1 on pulmonary surfactant synthesis of cultured alveolar type II cells]. AB - The effects of endothelin-1(ET-1) on pulmonary surfactant(PS) synthesis of cultured AT II cells were observed. The results showed that: 1. ET-1(10(-11)-10( 8) mol.L-1) enhanced PS synthesis of cultured AT II cells in a dose-dependent manner. 2. The minimum effective concentration of ET-1 enhanced PS synthesis of cultured AT II cells was higher than that of lung explants. 3. ETA antagonist BQ123 decreased the effect of ET-1 on PS synthesis, but ETB antagonist BQ788 did not change the effect. 4. ET-1 (10(-12) and 10(-10) mol.L-1) had no effect on the proliferation of AT II cells. These results confirmed that ET-1 can enhance directly and indirectly on the PS synthesis of AT II cells mediated via ETA receptor. The effect of ET-1 on PS synthesis was not induced by changing the number of AT II cells. PMID- 11938756 TI - [Two cases of clavicular tumor]. PMID- 11938757 TI - [A case of metastatic pituitary adenocarcinoma in cervical vertebra]. PMID- 11938758 TI - [A case of eutopic and ectopic pregnancy]. PMID- 11938759 TI - [A case of Budda-chiari syndrome diagnosed by ultrasonography]. PMID- 11938760 TI - [Differentially expressed cDNA sequences homologous with known genes in human nasopharyngeal carcinoma]. AB - In order to search the tumor suppressor genes correlated with pathogenesis of human nasopharyngeal carcinoma(NPC), we applied the PCR-based subtractive hybridization technique of representational difference analysis (RDA) to the primary culture cells of normal human nasopharyngeal epithelial and HNE1, a poorly differentiated NPC cell line. Following four successive subtractions of HNE1 complementary DNA from normal human nasopharyngeal epithelial cells complementary DNA, difference products were cloned into pGEM-T easy vector and nucleotide sequences determined. Comparison of cDNA sequences against the databases identified 9 known genes. Known genes included TRIP1(TGF beta receptor interacting protein), TAF, ezrin, MHC II, actinin, Histone H1 zero, Cytokeratin 13, Squalene Synthetase and RNA Synthetase-like. Some of them have an effective suppressive ability on the tumor. In this study, we have demonstrated that cDNA RDA is an effective strategy for systematically isolating differences in gene expression between two related but functionally distinct cells. Our results also indicated that the NPC includes interaction of multigenes and this experiment offers a new route for NPC research. PMID- 11938762 TI - [Effect of nitric oxide on HSV-1 infection of HeLa cells and Vero-E6 cells]. AB - The effects of nitric oxide(NO) on HSV-1 infection of HeLa cell line and Vero-E6 cell line were investigated in this study. The titers of HSV-1 in supernatants from infected culture of Vero-E6 cells were decreased by a NO donor, sodium nitroprusside(SNP), in a dose-dependent manner, while the control compound potassium ferricyanide (KFC) had no effect. However, SNP had no effect on that of HeLa cells. The results reaffirm the antiviral effect of NO and suggest that the susceptibility of different cell lines to the effect of NO might be varied. PMID- 11938761 TI - [Significance of human telomerase RNA expression in situ in malignant melanoma]. AB - By using the method of hybridization in situ, human telomerase RNA(hTR) are detected in 36 cases of malignant melanoma tissue, 16 cases of nevocellular nevus and 12 cases of normal skin. Results showed that the positive expression percentage of hTR in malignant melanoma, nevocellular nevus and normal skin was 86.3%, 6.26% and 12.7%, respectively. The positive expression percentage of hTR in malignant melanoma was significantly higher than that of nevocellular nevus and normal skin(P < 0.01). Meanwhile, the positive expression intensity of hTR in metastatic malignant melanoma is frequently stronger than that of original malignant melanoma(P < 0.05). The results suggest that hTR might indirectly reflect the telomerase activity, and be used as a new marker for diagnosis of malignant melanoma and metastasis prediction. PMID- 11938763 TI - [Influence of triiodothyronine and insulin on liver adenine nucleotide levels in portal triad-clamping rats]. AB - OBJECTIVE: To assess the influence of triiodothyronine(T3) and insulin on liver adenine nucleotide levels in portal triad-clamping rats. METHOD: A rat model of normothermic portal triad clamping was established to observe the influence of T3 and insulin on the contents of liver adenine nucleotides and survival rates. RESULT: The ATP levels and the 72 h-survival rate of T3 group were significantly higher than those of insulin group and saline group(P < 0.05). The blood glucose levels were obviously decreased and all the rats died within 6 hours postoperation in insulin group. CONCLUSIONS: T3 can improve hepatic energy metabolism after portal triad clamping and increase survival rate, whereas such effects were not found in insulin group. PMID- 11938764 TI - [Follow-up study of human herpesvirus 6 and human cytomegalovirus infection in organ transplantation recipients]. AB - OBJECTIVE: To trace the study of the prevalence and adverse factors of human herpesvirus 6(HHV-6) and human cytomegalovirus(HCMV) infection in organ transplantation recipients. METHOD: Blood and urine specimens were detected by virus isolation, polymerase chain reaction(PCR), and indirect immunofluorescent assay. RESULT: HCMV and HHV-6 isolation rate were 25.8% and 32.3%, respectively. Anti-HCMV IgM and anti-HHV-6 IgM positive rates before operation were 0% and 3.2%, their positive rates after operation were 19.4% and 25.8%, respectively. HCMV-DNA and HHV-6-DNA positive rates before operation were 35.5% and 45.2%; their positive rates after operation were 45.2% and 61.3%, and kept the high positive rates for a long period. The patients who died within 3 months were all complicated by HCMV and/or HHV-6 productive infection. The productive infection rates of HCMV and HHV-6 were 25.8% and 32.2%, respectively. CONCLUSION: The majority of HCMV and HHV-6 infection are reactivation. Productive infection attack at 2 weeks after operation, reaches the peak at 3-4 weeks after operation, and attacks at anytime within 3 months. HCMV and/or HHV-6 productive infection are fatal to recipients. PMID- 11938765 TI - [Methodological study on the assay of glycosylphosphatidylinositol-specific phospholipase D activity in serum]. AB - A method for the assay of glycosylphosphatidylinositol-specific phospholipase D(GPI-PLD) activity in human serum was established by using glycosylphosphatidylinositol(GPI) anchored placental alkaline phosphatase(PLAP) as a substrate. Serum GPI-PLD activity was determined both qualitatively and quantitatively by PEG/dextran and triton-X-114 partitioning respectively, and some kinetic properties of its enzymatic reaction were also studied. This method revealed a satisfactory reproductivity(CV = 3.6%) and a high sensitivity(2 microliters serum was sufficient). In addition, it is practical, economical, and feasible to be widely applied. The activities of GPI-PLD measured by this method in 100 normal subjects ranged from 30% to 50% (in terms of the percentage of converting GPI anchored PLAP substrate to the anchor-free product). PMID- 11938766 TI - [Platelet stereological study in patients with HFRS]. AB - To investigate the effects of stereological alteration of platelets on hemorrhage in hemorrhagic fever with renal syndrome(HFRS), platelet mean cross area(A), area density of alpha-granules(Na), volume density(Vv), the mean open tubule(OCS) and pseudopodia mean(M) were performed stereometric analysis in 12 patients with HFRS. The platelet A was observed, and no significant difference was found compared with the control(P > 0.05), while the number of pseudopodia M was higher, and the open tubule number was lower than those of the control(p < 0.01). The results suggest that in addition to the total platelet count decrement, the quality deterioration and functional activity abnormalities of platelets play an important role on the disordered hemostasis and coagulation in HFRS. PMID- 11938767 TI - [Experimental study on ultrashort wave therapy on the healing of fracture]. AB - The models of fracture in the rabbit radii were made. The rabbits in the experimental group were treated--with ultrashort wave at the fracture sites. The changes in the x-ray, histology, and biomechanics were observed dynamically. The results were that the speed and quality of the fracture healing in the experimental group were much better than in the control group. PMID- 11938768 TI - [Study of levels of antibody against coxsackievirus B in different adults in Changsha area]. AB - In order to verify the antibody levels against coxsackievirus B(CVB) in different adults, specific anti-CVB-IgM and anti-CVB-IgG have been detected in 46 viral myocarditis, 121 viral-hepatitis patients and 108 healthy donors by using immuno enzyme histochemical method, and the antibody titres were also tested. The results showed that the positive rate of anti-CVB in viral myocarditis group, viral hepatitis group and donor group were 80.4% (37/46), 25.6% (31/121) and 40.7% (44/108), respectively. The GMT of anti-CVB-IgM and anti-CVB-IgG in viral myocarditis group, hepatitis group and donor group were 1:84.84, 1:9.07, 1:64.00 and 1:179.27, 1:9.59, 1:21.42, respectively. The positive rate and GMT of anti CVB-IgM and anti-CVB-IgG in viral myocarditis group were significantly higher than those in the other two groups. The positive rate had no statistical difference between men and women. The results indicate that the majority of the healthy population in Changsha area is susceptible for CVB and most cases of the viral myocarditis many be related to CVB infection. It is considered that CVB can cause many diseases. Furthermore, there are no effective vaccine against CVB so far. One should think of the probability of CVB infection when encountering a patient. PMID- 11938769 TI - [Research on quality of life of stroke patients and linear model of its impact factors]. AB - The present study reviewed the investigation of 278 stroke patients of Changsha city population in 1994. The result showed that patients of stroke might have different quality of life during the process of their illness. Multiple regression model indicated that major factors on the chronic stroke patients were type of stroke, age, home nursing time, labor-losing days, occupations. The study provides reference source for prevention of stroke and improvement of quality of life. PMID- 11938770 TI - [A study of scavenging action of purslane aquenous extracts on oxygen free radical]. AB - Auto-oxidation in alkalesent solution with the help of pyrogallol can generate O2.-, and the system of 1, 10-phenathroline-Fe(II)/H2O2 can produce .OH. Spectropholometry (wave length 322 nm and 536 nm) was used to observe the effect of fresh purslane aqueous extracts and the purslane aquenous extracts processed by boiling water upon O2.- and .OH. The results showed that fresh purslane aqueous extracts could obviously help generate O2.- and reduce .OH, and the aqueous extracts of purslane processed by boiling water had a scavenging action on O2.- and .OH greatly. PMID- 11938771 TI - [PCR for the detection of 16S rRNA gene bacteria]. AB - According to the high conservative region of 16S rRNA gene in bacteria, PCR primers of the broad-range bacteria, gram-positive bacteria and gram-negative bacteria were synthesized to detect 13 bacterium species and (40) clinical specimens. All the tested bacterium species were positive. The lowest concentration of Escherichia Coli detected by serial dilution was 4 CFU.ml-1. The positive rate of PCR (27/40) was higher than that of bacterium culture(18/40). The results indicate that these PCR primers possess high specificity and sensitivity in identifying 16S rRNA gene of bacteria. PMID- 11938772 TI - [Long-term follow-up study on Hunan aqueous drainage implantation for refractory glaucoma]. AB - One hundred and three cases(106 eyes) of refractory glaucoma underwent Hunan aqueous drainage (HAD) implantation from July 1993 to January 1998. The mean period of postoperative follow-up was 32.8 months. The intraocular pressure(IOP) was decreased from preoperative 6.6 +/- 1.8 kPa to postoperative 2.2 +/- 1.4 kPa at the 1st year after surgery. The success rates were 83.8% at the 1st year, 80.4% at the 2nd year, 77.1% at the 3rd year, 71.0% at the 4th year, and 64.3% at the 5th year using Kaplan-Meier lifetable analysis. The neovascular glaucoma had lower success rate. The success rate in 38 patients who were used adjunctively with mitomycin C in the operation was 89.5% at the 1st year. The common complications included early continuous flat anterior chamber, local choroidal detachment, increased IOP, and so on. This study suggests that HAD implantation is an effective method in the treatment of refractory glaucoma in spite of its complications. PMID- 11938773 TI - [Enhanced cytotoxicity of VM-26 in human malignant glioma cells by calcium channel blocker nicardipine in vitro]. AB - Using the monolay tumor cell culture system and the method or MTT colorimetric analysis, the cytotoxicity of the anticancer drug VM-26 and Nicardipine(NCDP) in human malignant glioma cell line U251 was studied. The results showed that the cytotoxicity of VM-26 in U251 was enhanced by NCDP at a low dose which did not show direct cytotoxicity. The authors considered that VM-26 combined with NCDP can heighten the intracellular anticancer drug concentration, reverse the drug resistance and heterogeneity in human malignant glioma cells. The mechanism of the enhanced effect was also discussed. These findings will provide an alternative chemotherapeutic clue to treat the postoperative malignant gliomas. PMID- 11938774 TI - [Study on anticardiolipin antibody in the patients with primary nephrotic syndrome]. AB - Combination of the anticardiolipin antibody with the phospholipids on the surfaces of the endothelia and thrombocytes can damage the basic functions of cells. In this study, the anticardiolipin antibody was detected in 68 cases with primary nephrotic syndrome and 30 healthy persons. The positive rates were 32.35% and 3.33%, respectively. The hypercoagulability and glomerular impairment in the positive cases were more evident than those in the negative cases. In addition, the sensitivity to treatment in the positive cases was markedly decreased. These findings indicate that the anticardiolipin antibody may play an important role in pathophysiological processes of primary nephrotic syndrome. PMID- 11938775 TI - [Intestinal bacterial translocation in patients with biliary tract diseases]. AB - To determine whether intestinal bacterial translocation occurs in man, the authors cultured the tissues of mesenteric lymph nodes(MLNs), bile, portal and peripheral venous blood obtained prior to and during operation in 35 patients with biliary diseases. No positive peritoneal swab, portal blood and peripheral blood culture were shown. Thirteen of 20 patients with biliary obstruction (65%) had bacteria in their MLNs, while no positive MLN culture was found in 12 patients without biliary obstruction. The most common bacteria recovered from the MLNs were Gram-negative enteric bacilli. Thus, it is considered that extrahepatic biliary obstruction may induce intestinal bacterial translocation in man. PMID- 11938777 TI - [Electroencephalography and transcranial Doppler analysis in children with migraine]. AB - Electroencephalogram(EEG) and transcranial Doppler(TCD) recordings were carried out in 25 children with migraine during ictal and interictal period. The results were that the abnormal rate of EEG and TCD was 85% and 72%, respectively; the positive rate of EEG provoked test was 60%; there was the single untypical epileptiform wave in the occipital in 3 patients by EEG. We conclude that EEG combined TCD examination in migraine can increase the correct rate of diagnosis and help to differ epilepsy from migraine. PMID- 11938776 TI - [Efficacy of native carvedilol in patients with essential hypertension]. AB - To compare the efficacy of native carvedilol(Cv) with native atenolol(At), we treated 80 cases of essential hypertension(stage I-II) for 4 weeks, using both open-test and double blind randomized imitative methods. The daily dose of carvedilol was 10 mg to 40 mg, and that of atenolol was 50 mg to 100 mg. Both drugs were given once daily. Out of 30 cases of Cv group, carvedilol showed significant efficiency in 20 cases, and was effective in 6 cases. The total therapeutic efficiency was 86.2% (26/30). In the double blind groups(25 patients in each group), those treated with carvedilol(Group A) had 21 effective cases(84.0%) while those treated with atenolol(Group B) had 18 effective cases(72.0%). A lower incidence of adverse effects was observed in carvedilol group. Dizziness occurred similarly in both drugs, but it lasted only a short time and could be tolerated. The recommended dose of 10-30 mg once daily would be appropriate. After treatment there was a significant reduction in blood pressure in both groups(P < 0.01). It suggests that native carvedilol is a safe and effective anti-hypertensive agent. PMID- 11938778 TI - [No arresting aorta undergoing open heart surgery with CPB through a right anterolateral thoracotomy]. AB - We performed operation of CPB without arresting aorta on 57 cardiac patients through a right anterolateral thoracotomy. Results showed: except that one case of ASD required a 2nd time thoracotomy to hemostasis for a lot of drainage, another case of residual shunt postoperation of TOF was not satisfactory in repair, the others are perfect. There were no operative or hospitalized mortality, no incision infection. It has also solve the psychological distress of female patients provoking by midline showy scar of median sternotomy, so it has acquire a good surgical cosmetic effect. PMID- 11938779 TI - [Changes on positive rate and distribution of Helicobacter pylori during progression of gastric cancer]. AB - We observed changes on the positive rate and the distribution of helicobacter pylori (HP) in 7 case during early stage of gastric cancer and in 42 cases during middle-late stage of gastric cancer. The results showed that 1. the positive rate during early stage of gastric cancer was 57.1%, the positive rate during middle rate stage of gastric cancer was 23.8%, 2. HP was not found within the gastric cancer lesions, 3. for HP positive cases, HP distribution during early stage of gastric cancer was more sparse than during middle-late stage of gastric cancer. These results suggested that HP can hardly live within the gastric cancer tissue, so the progression of gastric cancer is probably independent of HP. It may be a question that HP eradication can prevent later development of gastric cancer. PMID- 11938780 TI - [Determination of TNF-alpha and endotoxemia in patients with chronic liver diseases]. AB - The incidence of endotoxemia in 153 patients with different liver diseases was determined by Limulus Lysaite Test(LLT). Comparative studies were carried out on the levels of tumour necrosis factor-alpha(TNF-alpha) between positive and negative endotoxemia in patients suffering from chronic liver diseases. At the same time, 58 patients of positive endotoxemia were divided into two groups- bifidobiogen group (38 cases) and control group(20 cases). The patients in bifidobiogen group were treated mainly with daily administration of bifidobiogen for 1 month besides general liver--protection therapy and the bifidobiogen was not given to the control group. The results showed that: 1. Incidence of endotoxemia in 153 patients suffering from various types of liver diseases was 65.36%; the highest rate was found in the patients with severe hepatitis; 2. The TNF-alpha level was obviously increased in the patients of positive endotoxemia; 3. The total efficacy in bifidobiogen group was significantly higher than that in control group. The data suggest that chronic liver diseases are generally complicated with endotoxemia, and TNF-alpha plays an important role in the pathogenesis of endotoxemia. The serum endotoxin level may be reduced after the treatment with bifidobiogen. PMID- 11938781 TI - [Serum lipids, lipoprotein and stroke]. AB - The serum levels of lipids and lipoproteins were measured in 180 consecutive patients with acute stroke(90 hemorrhagic stroke, 90 ischemic stroke) and 107 age and sex-matched controls. As compared with the control group, hemorrhagic group had lower levels of total cholesterol(TC) (P < 0.05), low density lipoprotein cholesterol(LDL-C) (P < 0.01) and Lp(a) (P < 0.05) while the ischemic group had higher serum TC(P < 0.05), LDL-C(P < 0.01), Lp(a) (P < 0.01) and lower high density lipoprotein-cholesterol(HDL-C) (P < 0.05) and ApoAI(P < 0.01). Multiple factor logistic regression showed that cigarette smoking, hypertension and family history of stroke were the common risk factors for both hemorrhagic and ischemic stroke, and that lowered TC and elevated LP(a) levels were independent risk factors for hemorrhagic stroke and ischemic stroke respectively. PMID- 11938782 TI - [Measurement of soluble Fas in patients with hematological malignancy]. AB - We measured serum levels of soluble Fas(sFas) in 68 patients with hematological malignancy by using sandwich ELISA and detected bone marrow mononuclear cell membrane Fas(mFas) positive cells in 46 patients with acute leukemias by immunohistochemical technic. It was found that sFas levels were significantly higher in acute lymphoblastic leukemia(ALL) patients(median 10.48 +/- 5.89 ng.ml 1) and non-Hodgkin's lymphoma(NHL) patients(median 18.26 +/- 16.47 ng.ml-1) than those in healthy donors (median 0.92 +/- 0.88 ng.ml-1) (P < 0.05). Elevated sFas levels were correlated with the disease stage and progress, but not with other clinical parameters. According to the follow-up of five ALL patients, their serum sFas levels decreased after complete remissions. We conclude that sFas has prognostic value and is an effective clinical parameter with ALL or NHL patients. In addition, sFas levels in the sera of acute non-lymphoblastic leukemia patients were also found to be unchanged compared with those in the normal control. PMID- 11938783 TI - [The relationship between surgical staging, pathologic grading, operative type and postoperative recurrence in giant cell tumor of bone]. AB - Sixty-eight surgical procedures of 58 patients with Giant Cell Tumor of Bone(GCT) were followed up over two years. The recurrence rate of GCT was Stage 1: 0 in 5, Stage 2: 15.4% in 39 and Stage 3: 37.5% in 24. It was significantly higher in Stage 3 than in Stage 2 (P < 0.05). The recurrence rate of GCT in Stage 2 was 30.8% in 13 intralesional curettage, and 0 in 17 wide or radical resection(P < 0.05). We conclude that the surgical staging of GCT has important value in predicting postoperative recurrence, in accurately evaluating the operative efforts and in guiding treatment, and that the operative type is a significant factor effecting on the recurrence of GCT. PMID- 11938784 TI - [Investigation of the bone mineral density in hemodialysis patients for different terms]. AB - The investigate the change of bone mineral density(BMD) in hemodialysis patients for different terms of hemodialysis, 50 patients were divided into three groups according to the duration of hemodialysis. Group I was < 5 years. Group II was 5 10 years, and Group III > 10 years. The BMD of the patients were assayed by using dual energy X-ray bone absorptiometer. Serum calcium, phosphorus, aluminium, and intact parathyroid hormone(iPTH) were detected at the same time. The mean BMD and serum iPTH were 0.624 +/- 0.180 g.cm-2 and 9.67 +/- 9.46 ng.cm-1 in Group I, 0.573 +/- 0.160 g.ml-2 and 10.16 +/- 28.05 ng.ml-1 in Group II, and 0.513 +/- 0.117 g.cm-2 and 10.75 +/- 24.94 ng.ml-1 in Group III, respectively. There was no statistical significance in BMD (P > 0.05) and iPTH(P > 0.05) among the above groups. The BMD of hemodialysis patients is not affected by different terms of hemodialysis. PMID- 11938785 TI - [Expression of bcl-2, bax and p53 protein in the basal cell carcinoma]. AB - The expression of bcl-2, bax and p53 protein was studied in the basal cell carcinoma with 32 patients by immunohistochemical method. The results showed that bcl-2 protein was positive in 26 patients (81.2%), bax protein was positive in 9 patients(28.1%), and p53 protein was positive in 13 patients (40.6%); there is a negative correlation between the expression of bcl-2 and bax protein(P < 0.05). These findings suggest that apoptosis is suppressed which might cause the production of basal cell carcinoma. PMID- 11938786 TI - [Analysis of complications and deaths in aged patients with obstructive jaundice]. AB - The reasons of complications and deaths in 98 aged patients with obstructive jaundice were analysed in the paper. Among them, 52 cases were diagnosed as benign obstruction, 46 cases as malignant obstruction; 92 patients were treated by surgical management(15 died), 6 patients were treated without operation(4 died). The results showed that effective management on time and intensive perioperative care are important to minimize the mortality rate. Malignant obstructive jaundice was more harmful to the patients' renal function no the mortality rate would be increased. The operations, internal drainage and pressure reduction of biliary duct, were performed on time, thus the survival rate was improve. PMID- 11938787 TI - [Urotoxic effect to intestinal bacteria in patients with chronic renal failure]. AB - To analyse the differences of intestinal bacterial quality and quantity, the feces taken from chronic renal failure(CRF) and healthy subject were cultured aerobicly. Then the isolated E. coli from CRF patients and healthy subjects was inoculated respectively into cultural medium with CRF serum and incubated whose concentration of UN, Cr and UA in medium before and after incubation was measured to observe the bacteria degrade of the above-mentioned urotoxin. RESULT: The intestinal E. coli can apparently degrade UN, Cr and UA in CRF patients which suggests that urotoxin may induce E. coli variation. PMID- 11938788 TI - [Influence factors on correct diagnosis of non-epileptic paroxysmal disorders in childhood]. AB - To investigate the influence factors on correct diagnosis of non-epileptic seizures(NES) in childhood, we analyzed 24 h ambulatory electroencephalography(AEEG) and clinical features of the attacks in 159 children with NES. The results showed that 28% of all patients were identified as NES plus epileptic seizures. Epileptiform EEG discharges were observed in 10% of pure NES children. Abnormal findings on neurodiagnostic evaluation were obtained in 32% of NES plus epileptic seizure groups, in 11% of pure NES groups. Unnecessary anti epileptic drugs were received in 18% of pure NES children and the kind or doses of the medicines were increased in 50% of NES plus epileptic seizure children who had been mistaken for refractory epilepsy. The influence factors on correct diagnosis of NES are mainly; variability of non-epileptic seizure expression, interictal epileptiform EEG discharges, neurologic abnormal findings, and anti convulsant abuse. PMID- 11938789 TI - [The phasic and regional variations of lipid peroxidation after diffuse brain injury in rats]. AB - OBJECTIVE: Using a diffuse brain injury animal model, we studied the time course of lipid peroxidation in different regions of injured rat brains. METHODS: 53 male SD rats were randomly divided into sham, post-traumatic 1, 2, 4, 7, 24 h groups (n = 8) to measure the MDA (malondialdehyde) at the bilateral frontal, temporal, parietal, striatum and brain stem, another 5 rats were researched for histological study. RESULTS: At the frontal, parietal and brain stem, the MDA levels were 36.7%, 41.8% and 35.1%, respectively, higher than sham at one hour after injury (P < 0.01). The MDA levels in these regions continued to increase and peaked at 4 hours after the injury. The levels decreased slowly, and by 24 hours, they were still significantly higher than the sham control's. The elevation of MDA levels was less in the striatum and the temporal compared with above regions at one hour. Their rising rates were 16.9% and 13.3%, respectively(P < 0.01). The MDA levels in there two regions continued to increase even after 4 hours of injury, but the degree of elevation never exceeded 35%. CONCLUSION: There is an immediate, post-traumatic burst of MDA production, suggesting the formation of free radicals after diffuse head injury. Even though all the regions sampled show the same effect, certain regions are less affected in this diffuse brain injury animal model. PMID- 11938790 TI - [Study of BAEP, TCD and neck rotation test in vertebrobasilar-artery insufficiency patients]. AB - Forty patients with vertebrobasilar-artery insufficiency(VBI) were checked by BAEP and BAEP together with neck-rotation test. Their positive rates were compared with those of TCD. The results showed that: 1. the positive rate of BAEP was 50%, lower than that of TCD; 2. after neck rotation test, the positive rate of BAEP increased from 50% to 86.3%. The results suggest that BAEP together with neck rotation test can obviously increase the positive rate of BAEP. The method is simple and effective in clinical application. PMID- 11938791 TI - [Clinical observation of misoprostol on induction in late pregnancy]. AB - Eighty late pregnancy women were randomized into two groups. Misoprostol group and oxytocin group. We observed the different effects of misoprostol and oxytocin on induction, the different effects of misoprostol on induction which was given through rectum or through vagina. The results indicate that the rate of successful induction in misoprostol group is higher than that of oxytocin group, the rate of caesarean section in misoprostol group is lower than that of oxytocin group. There was no significant difference in the durations of misoprostol which was given through rectum or through vagina on induction. It is suggested that misoprostol is more effective on induction than that of oxytocin. We can choose misoprostol given through rectum to prompt delivery in conditions such as premature rupture of membranes and vaginitis. PMID- 11938792 TI - [Examination of human herpesvirus-6 antibody in blood patients serum]. AB - To know the infection state of human herpesvirus-6(HHV-6) in leukemia, lymphoma, myeloproliferative disorder syndrome, and multiple myeloma, indirect fluoroimmunoassay was taken to examine the rate of HHV-6's antibody. The results were that: the patient's rate of HHV-6 antibody was 75.6% (149/197), the blood donor's rate was 45.6% (115/252), there was a significant difference between them (P < 0.01). The results show that the examined patients are infected by HHV-6, which has a good relationship with leukemia, lymphoma, myeloproliferative disorder syndrome, and multiple myeloma. PMID- 11938793 TI - [Application of RF-II pedicle screw system in lumbar spondylolisthesis]. AB - Thirty-eight patients with isthmic(31 cases), degenerative(6 cases), and post traumatic(1 case) spondylolisthesis were treated with RF-II pedicle screw system. Eighteen patients were classified as Degree I lesion, 14 as Degree II lesion, and 6 as Degree III lesion. Postoperatively, 34 cases(90%) obtained an anatomical improvement obviously. The percentage of the patients who got excellent and good recovery was above 90%. We conclude that RF-II pedicle screw fixation system has the strongest three-dimensional rigidity and, attains complete and satisfactory decompression. The entire intervertebral disk in the level of the lesion is suggested to be removed. The system is ingeniously and originally designed, and is easy to obtain a simple and an effective reduction of the spondylolisthesis. Additionally, the application of the system can obtain a good interbody fusion by three steps: broadening the interbody space; stuffing the space with broken bones; and compressing the involved vertebral body. The authors considered that the RF-II screw system is one of the most ideal internal fixations for the treatment of lumbar spondylolisthesis. PMID- 11938794 TI - [Hip ambulatory abduction in the treatment of Legg-Calve-Perthes disease]. AB - Forty-eight cases of Legg-Calve-Perthes disease were treated with hip ambulatory abduction brace. The children were able to walk, sit, climb, or squat with the brace. Thirty-nine cases were followed up more than three years. The total excellent rate was 82%. The result of Catterall Grade I-II cases was all excellent. The excellent rate was 80% below five-year-old cases of Catterall III IV during the start of the treatment and 66.6% above five. With the active hip abduction and internal rotation, the femoral head was held well. The results show that the method is simple and with no operation and with satisfactory effect of the treatment. PMID- 11938795 TI - [Determination of annaka injection by an improved double wavelength spectrophotometry]. PMID- 11938797 TI - [Analysis of 56 cases of generalized juvenile rheumatoid arthritis]. PMID- 11938796 TI - [Relation between Chlamydia pneumoniae infection and acute attack of asthma]. PMID- 11938798 TI - [Changes of plasma endotoxin and nitric oxide in patients with liver cirrhosis]. PMID- 11938799 TI - [Influence of chemotherapeutants and cytokines on growth and transgene expression of bone marrow cells from MT/p210(bcr-abl) transgenic mice]. AB - The bone marrow cells of CML(chronic myelogenous leukemia) transgenic mice carrying metallothionein(MT) promoter/enhancer, bcr-abl(p210) cDNA and SV40 splicing/poly(A) signal sequences were cultured in liquid and soft agar with hydroxyurea (Hu), 5-fluorouracil(5-Fu), mouse stem cell factor (mSCF) and mouse interleukin-3(mIL-3) independently or combinatively. The cell or colony counting and transgene RT-PCR analysis showed that the proliferation, colony formation and transgene expression of the bone marrow cells were advanced in the combinative culture of mSCF with mIL-3, but they were significantly inhibited in the culture without any growth factors, or with mSCF, mIL-3 and Hu or 5-Fu. These findings suggest that the combinative utilization of chemotherapeutants and cytokines is a potentially effective strategy of the clinical treatment for CML. PMID- 11938800 TI - [Affection of epidermal growth factor on VIP secretion and VIPR expression in airway epithelial cells]. AB - In order to confirm the essential role of vasoactive intestinal peptide(VIP) on protection of local airway epithelium and investigate the modulation of the protective effect of VIP, secretion of VIP and expression of VIP receptor(VIPR) were studied in bronchial epithelial cells(BEC), and alveolar macrophages (AM). The radioimmunoassay was used for determining VIP secretion and the radio-ligand affinity analysis for VIPR expression. The effect of epidermal growth factor(EGF) was observed. The results showed that: 1. Both BEC and AM secreted VIP basically without remarkable stimulus, and some vesicles with electron-dense core which usually exist specifically in neuropeptide secreting cells were found in plasma of BEC; 2. In a dose-dependent manner, EGF promoted BEC to secrete VIP(r = 0.84, P < 0.05), and it was found that VIP secretion from AM was also elevated in EGF pretreated group(P < 0.05); 3. EGF exhibited a dose-dependent up-regulatory effect on the expression of VIPR in BEC(r = 0.95, P < 0.01). These results indicate that VIP may play an essential role in the protection at local airway by autocrine or paracrine, and the protective effect can be modulated by growth factors. PMID- 11938801 TI - [Methods for making human sectional specimens]. PMID- 11938803 TI - [Chinese drugs for invigorating spleen to remove dampness, activating blood circulation to eliminate turbid in retarding progression of chronic renal failure]. AB - OBJECTIVE: To investigate the effect of Chinese drugs (CD) for invigorating Spleen to remove dampness, activating blood circulation to eliminate Turbid in retarding progression of chronic renal failure (CRF). METHODS: Thirty-nine patients with CRF were divided into two groups randomly, 18 patients in the control group were treated with low protein diet and controlling blood pressure and 21 patients in the CD group were treated similarly with that of the control group but with additional CD. Levels of serum creatinine (SCr), blood urea nitrogen (BUN), blood albumin (Alb), cholesterol (CH), triglyceride (TG), high density lipoprotein (HDL) and hemoglobin (Hb) were checked every 2 months, and the rate of progression of CRF was estimated by slope of the creatinine reciprocal (1/SCr) with time (month). RESULTS: Levels of SCr and BUN in the CD group were significantly lower and HDL markedly higher than those in the control group, P < 0.05. Mean slopes of the creatinine reciprocal with time in the two groups was significantly different, P < 0.01. CONCLUSION: Additional CD treatment based upon the protein diet and controlling blood pressure could retard the progression process of CRF evidently. PMID- 11938802 TI - [Assessment on application of Chinese herbs in process of corticosterone reduction in nephrotic syndrome]. AB - OBJECTIVE: To evaluate the effect of Chinese herbs in the process of corticosterone (CS) reduction in nephrotic syndrome. METHODS: One hundred and twenty patients of idiopathic nephrotic syndrome sensitive to prednisone treatment were divided into two groups randomly. The dosage of prednisone used was reduced by routine in both groups but Chinese herbal treatment was added to the treated group in the period of prednisone reduction. The effect of Chinese herbs was estimated by observing the recurrence rate of disease and side-effects of prednisone. RESULTS: The recurrence rate of disease and the occurrence rate of side-effects of prednisone in the treated group were lesser than that in the control group significantly (P < 0.01). CONCLUSION: Additional Chinese herbs in the process of CS reduction in the treatment of CS sensitive nephrotic syndrome could extenuate markedly the recurrence rate of disease and the side-effect of CS reduction. PMID- 11938804 TI - [Clinical study on Shenning Mixture in treating IgA nephropathy]. AB - OBJECTIVE: To study the clinical therapeutic effect of Shenning Mixture (SNM) in treating IgA nephropathy. METHODS: Patients were treated separately with SNM or prednisone by randomized grouping. The criteria, including clinical symptoms, signs, hematuria, albuminuria and immune globulin were observed. RESULTS: The total effective rate and the complete remission rate in the SNM group were 97.1% and 45.7% respectively, while in the control group, they were 37.1% and 8.6% respectively. Comparison of the two groups showed that the therapeutic effect in the former was better than that in the latter significantly, P < 0.01. The hematuria and albuminuria extenuated and serum IgA lowered more significantly in the SNM group than those in the control group (P < 0.05, P < 0.01). CONCLUSION: SNM has obvious therapeutic effect on the hematuria and albuminuria in treating IgA nephropathy. PMID- 11938805 TI - [Clinical observation on operation of cerebral speech area under combined acupuncture-drug anesthesia]. AB - OBJECTIVE: To study the effect of combined acupuncture-drug anesthesia (CADA) in neurosurgical operation of cerebral speech area. METHODS: Twenty-five patients with tumor close to the speech area underwent neurosurgical operation under CADA or general anesthesia were observed, and the post-operational language function protecting effect was analyzed. RESULTS: Among the 15 patients accepted CADA, total tumor resection was achieved in 9 cases, and subtotal or partial resection achieved in 6 due to their tumor too close to the speech area. No post operational complication such as dysphasia or aggravation of functional disorder was found in the patients. In the 10 cases underwent general anesthesia, 7 had tumor total resection, 3 had partial resection, and 3 cases suffered from post operational dysphasia. CONCLUSION: CADA is helpful in speech area judgement in neurosurgical operation so as to avoid the operational injury on functional area. It is an important method of anesthesia in surgical operation on cerebral speech area. PMID- 11938806 TI - [Clinical study on treatment of nasopharyngeal carcinoma by radio- and chemotherapy with supplementary moxibustion on Shenque point]. AB - OBJECTIVE: To evaluate the effect of supplementary moxibustion in treating III, IV a stage nasopharyngeal carcinoma (NPC) with radio- and chemotherapy. METHODS: Fifty-six cases of NPC were randomly divided into two groups, 28 in each group. They were treated with radiotherapy in routine or chemotherapy adopting AD protocol. Salt-separated moxibustion on Shenque (Ren 8) point was given to the treated group from beginning of radio- and chemotherapy for 30 times as one therapeutic course. RESULTS: The remission rate in the two groups after radio- and chemotherapy was not different significantly. The toxic and side-effect occurrence was less in the treated group than that in the control group (P < 0.05). The 5-year local control rates of NPC and cervical lymphnode were 85.7% and 85.0% in the treated group, which were higher than those in the control group (78.6% and 78.9%). The 5-year survival rate in the two groups were 50.0% and 35.7% respectively. After radio- and chemotherapy, the blood content of malonyldialdehyde (MDA), middle molecular substance and sulfhydryl reduced the SOD activity ascended in the treated group, the difference was significant as compared with those in the control group (P < 0.05, P < 0.01). CONCLUSION: The supplementary moxibustion on Shenque point could obviously reduce the toxic side effect of advanced NPC patients treated with radio- and chemotherapy. PMID- 11938807 TI - [Effect of qi replenishing and blood circulation activating drugs in treatment of middle-advanced pancreatic cancer with radio- and chemotherapy]. AB - OBJECTIVE: To study the effect of Qi replenishing and blood circulation activating drugs (QRBCAD) in treatment of middle-advanced pancreatic cancer with radio- and chemotherapy. METHODS: Patients were divided randomly into two groups, 28 patients treated with radiotherapy and intervention treatment as Group A and 30 patients treated with the same therapy but with additional QRBCAD medication as the Group B, and the effects in the two groups were compared. RESULTS: The short-term effective rate in Group A and B was 53.6% and 67.6% respectively (P < 0.05). The remission rates of jaundice and abdominal pain in Group A were 57.1% and 50.0% respectively, while in Group B, 83.3% and 76.7% respectively, there was significant difference between the two groups, P < 0.05. The gastroenteric reaction occurred in Group B was significantly lesser than that in Group A, P < 0.05. The 1- and 2-year survival rate in Group A were 50% and 21.4%, and in Group B, 80.0% and 46.6% respectively, significant difference (P < 0.05) showed in comparison of the two groups. CONCLUSION: QRBCAD could alleviate the gastroenteric reaction caused by radio- and chemotherapy, raise the clinical symptomatic effective rate of treatment, and prolong the 1- and 2-year survival rate of patients. PMID- 11938808 TI - [Clinical and experimental study on treatment of bronchial asthma with pingchuan oral liquid]. AB - OBJECTIVE: To observe the clinical effect of Pingchuan oral liquid, including Hanchuan oral liquid (HCOL) and Rechuan oral liquid (RCOL), and to explore its mechanism. METHODS: HCOL and RCOL were prepared based on the therapeutic principles of eliminate phlegm, remove blood stasis, lower reversed Qi flow and relieve asthma. HCOL was used to treat 50 patients of bronchial asthma in Group A and RCOL for 55 patients in Group B, and they were compared with 52 patients treated with Guilong Kechuanning capsule in the control group. The relevant animal experiment was also conducted. RESULTS: The total effective rate of HCOL was 88.0%, that of RCOL was 90.9%, they were significantly higher than that of the control (73.1%, P < 0.05). Moreover, the two new preparations showed the effects in easing main symptoms and signs of asthma, decreasing peripheral eosinophilic granulocyte count and immunoglobulin level, and improving pulmonary function superior to those of Guilong Kechuanning capsule (P < 0.05, P < 0.01). Experimental study showed that they could lower blood thromboxane B2 and endothelin, raise blood nitric oxide and 6-keto-prostaglandin F1 alpha. CONCLUSION: Pingchuan oral liquid is obviously effective in treating bronchial asthma, the mechanism possibly lays on adjusting immune function, anti-allergy and antagonizing inflammatory media. PMID- 11938809 TI - [Clinical and experimental study on treatment of acute catarrhal otitis media with eryanling oral liquid]. AB - OBJECTIVE: To study the clinical effect and mechanism of Eryanling (EYL) oral liquid in treating acute catarrhal otitis media (ACOM). METHODS: Sixty-eight cases (89 ears) of ACOM in the treated group were treated with EYL and compared with 34 cases (44 ears) in the control group treated with cephalexinum. Experimental study of effect of EYL on immune function in mice and non suppurative otitis guinea pig was also conducted. RESULTS: The total effective rate of the treated group and the control group was 91.0% and 84.1% respectively, and their rate of curing 80.9% and 70.5% respectively, though the effect in the former was better, statistic analysis showed no significance between them. The effect initiated obviously earlier in the treated group than that in the control group. Results of experimental study suggested that EYL could strengthen the nonspecific immune function, cellular immune function and humoral immune function in mice, and reduce the degree of inflammatory exudation and mucosa swollen in guinea pig. CONCLUSION: EYL has good therapeutic effect in treating ACOM. PMID- 11938810 TI - [Observation on curative effect of acute ischemic cerebrovascular disease treated with different dosage of ligustrazine]. AB - OBJECTIVE: To compare clinical curative effect of acute ischemic cerebrovascular diseases (ICVD) treated with different dosage of ligustrazine and to observe the effect of ligustrazine on hemorrheology. METHODS: Sixty-nine patients with ICVD were randomized into three groups, the Group A, B and C treated with 120 mg, 240 mg and 480 mg of ligustrazine respectively, the clinical curative effect, the changes in nerve function deficit score and hemorrheologic parameters before and after treatment were observed. RESULTS: The clinical curative effect and improvement of hemorrheologic parameters in Group C were better than those in Group A and B significantly (P < 0.05, P < 0.01 respectively). Ligustrazine also showed obvious effect in lowering blood level of fibrinogen in Group C. CONCLUSION: Treatment of acute ICVD with large dose of ligustrazine is good in improving clinical effect and hemorrheology, and without side effect basically. PMID- 11938811 TI - [Effect of yimai jiangya extract on plasma neuropeptide Y level in patients of senile hypertension with qi-deficiency and blood stasis syndrome]. AB - OBJECTIVE: To observe the effect of Yimai Jiangya extract (YMJYE) on plasma neuropeptide Y (NPY) level in patients of senile hypertension at stage II with Qi Deficiency and blood stasis Syndrome (QDBSS). METHODS: Sixty-eight patients were randomly divided into two groups, 36 patients in the treated group treated with YMJYE and 32 patients in the control group treated with captopine. Radioimmunoassay was used to examine the level of plasma NPY before and after treatment in the two groups. RESULTS: Before treatment, plasma NPY level was significantly higher in both groups than that in the old healthy persons (P < 0.01), it lowered significantly after treatment and the decrement was more obvious in the treated group in comparing with that in the control group (P < 0.01). Blood pressure was significantly lowered in both groups after treatment (P < 0.01) but with no significant difference between them. CONCLUSION: Plasma NPY level was increased in patients of senile hypertension with QDBSS. YMJYE had good effect in lowering plasma NPY level and reducing blood pressure. It is presumed that the NPY lowering effect of YMJYE might be one of its mechanisms in lowering blood pressure. PMID- 11938812 TI - [Study on the regulatory effect of Chinese herbal medicine on peritoneal lymphatic stomata and in enhancing drainage of ascites in mice with liver fibrosis]. AB - OBJECTIVE: To observe the regulatory effect of Chinese herbal medicine on peritoneal lymphatic stomata in carbon tetrachloride induced mouse model of liver fibrosis and its significance in treating ascites. METHODS: (1) To form the mouse model of liver fibrosis by intragastric administration of carbon tetrachloride. (2) Two kinds of Chinese herbal compound prescriptions were given separately by gastrogavage to observe their effect on prevention and treatment of liver fibrosis. (3) Hematoxylin and eosin and van Gieson staining were used in the observation of pathology and histology. (4) Scanning electronmicroscope and computer image processing were used. (5) urinary volume and sodium ion concentration were measured. RESULTS: The fibrotic change of the mouse models was alleviated with using both the prevention or treatment groups of the two compound prescriptions. The diameter of lymphatic stomata enlarged with increased number of opening and density of distribution. Urinary volume and sodium ion excretion increased after treatment. The effects displayed more significant in the group treated by compound prescription. CONCLUSION: Both kinds of Chinese herbal compound prescriptions have marked effect in alleviating liver fibrosis, regulating peritoneal lymphatic stomata, improving the drainage from the peritoneal cavity, causing increase of urinary volume and sodium ion excretion to reduce the water and sodium retention, and thus have favorable therapeutic effect in treating ascites. PMID- 11938813 TI - [Effect of xiongshao capsule on proliferation and apoptosis of vascular smooth muscle cells in rabbits observed by serum pharmacological method]. AB - OBJECTIVE: To observe the effect of Xiongshao Capsule (XSC) on proliferation and apoptosis of aortic smooth muscle cells (SMC) of rabbits by serum pharmacological method. METHODS: SMC were isolated from thoracic aorta of the normal rabbits and passaged. The SMC proliferation model was established by endothelin (ET). The medicated serum was prepared after given the normal rabbits suspension of XSC through gastrogavage for 10 days. The proliferation and apoptosis of SMC were observed by methyl thiazolyl tetrazolium (MTT) colorimetry, flow cytometry and agarose gel electrophoresis. RESULTS: ET obviously promoted proliferation of SMC, and the serum medicated with XSC could lower the proportion of SMC in S + G2 phase and markedly inhibited the proliferation of SMC caused by ET in dose dependent manner. It also had certain effect on inducing apoptosis of SMC. CONCLUSION: XSC could markedly inhibit the proliferation of SMC caused by ET and induce apoptosis of SMC, which might be the mechanism of preventing coronary restenosis after interventional treatment. PMID- 11938814 TI - [Experimental study on treatment of chronic renal failure with yishen qudu capsule]. AB - OBJECTIVE: To study the mechanism of action of Yishen Qudu capsule (YSQDC) in experimental rat model of chronic renal failure (CRF). METHODS: Membranous nephropathy model was induced by cationized bovine serum albumin intravenous administration in rats for 7 weeks and developed to CRF. The model was then treated with YSQDC of large or small dose by gastrogavage for 5 weeks. Oxidated starch was used in the control group. RESULTS: YSQDC could reduce proteinuria, serum creatinine, blood urea nitrogen, total cholesterol, triglyceride and serum circulating immune complex (P < 0.01 or P < 0.05), it also reduced the kidney coefficient, glomerular cells and tubular protein cast significantly (all P < 0.01). The strength of immune fluorescence weakened remarkably (P < 0.01). The pathological lesions improved significantly after treatment under light and electron-microscopic examination. CONCLUSION: YSQDC could improve renal function significantly and inhibit the progression of CRF. PMID- 11938815 TI - [Regulatory effect of Erbao granules on brain-gut peptide in juvenile animal model of anorexia]. AB - OBJECTIVE: To study the regulatory effect of Erbao granules (EBG) on central and peripheral brain-gut peptide in juvenile animal model of anorexia. METHODS: Juvenile rat model of anorexia was established by imitating the major cause of infantile anorexia and treated with EBG. The cholocystokinin-octapeptide (CCK-8) and beta-endorphin (beta-EP) concentration in hypothalamus, antrum pyloricum and peripheral blood were examined by radioimmunoassay. RESULTS: CCK-8 concentration in hypothalamus and plasma in the model rats increased (P < 0.05), while blood beta-EP concentration decreased (P < 0.05). After EBG treatment, the CCK-8 concentration normalized and beta-EP increased significantly. CONCLUSION: EBG could reduce the central and peripheral CCK-8 and increase beta-EP secretion significantly in the juvenile anorexia model. PMID- 11938816 TI - [Experimental study on effect of bushen qingli huoxue treatment on glomerular extracellular matrix of remnant kidney in rats]. AB - OBJECTIVE: To study the effect of Bushen Qingli Huoxue (BQH) treatment on glomerular extracellular matrix (ECM) and in retarding glomerular sclerosis in rats and to explore the mechanism and key of regulation. METHODS: Animal model was established by 5/6 nephrectomy and the study was carried out by using immune histochemical and pathological methods. Treatment of BQH started 1 week after operation. RESULTS: After being treated with BQH, the serum creatinine (SCr), blood urea nitrogen (BUN) and urinary protein excretion reduced markedly. Pathological examination showed accumulation of ingredients of glomerular ECM such as type I, IV collagen and laminin decreased significantly, the number of glomerular cells and proliferation cell nuclear antigen positive cells also decreased obviously, and showed that the glomerular sclerosis was retarded. The effect of BQH was similar to that of Enapril, but superior to that of Enapril in reducing BUN and SCr. CONCLUSION: BQH has the effects of reducing ECM accumulation, improving renal function and retarding glomerular sclerosis. PMID- 11938817 TI - [Ethological examination of Alzheimer's disease model rats treated with bushen yizhi decoction]. AB - OBJECTIVE: To explore the ethological improving effect of Bushen Yizhi Decoction (BSYZD) in treating model rat of Alzheimer's disease (AD). METHODS: Ethologic examination was carried out with Morris water maze on young, aged, AD model, Huperzine A treated and BSYZD treated groups of rats. RESULTS: In the BSYZD treated group, the escape latency (EL) curve placed between that of the young, aged and AD model, Huperzine A treated group; the percentage of middle circle swim path, the 40 cm circle swimming time, and the platform-crossing times were not different from those in the normal aged group, but were different significantly as compared with the AD model group, P < 0.05. CONCLUSION: BSYZD could ameliorate the behavior deficits of AD model. PMID- 11938818 TI - [Integrated traditional Chinese and Western medical research on neurosurgery]. PMID- 11938819 TI - [Mechanism of tongue figure formation]. PMID- 11938820 TI - [Mechanism of Rhubarb in preventing the occurrence of gastrointestinal function failure]. PMID- 11938821 TI - [Progress in the treatment of depression with new electroacupuncture]. PMID- 11938822 TI - [21st century--the era of integrated traditional Chinese and Western medicine marching to post-genome]. PMID- 11938823 TI - [Clinical study on tangweikang capsule in treating diabetic nephropathy]. AB - OBJECTIVE: To observe the effect of Tangweikang capsule (TWKC) in treating diabetic nephropathy (DN). METHODS: The 119 patients enrolled were divided into 2 groups, 78 patients in the TWKC group treated with TWKC and 41 patients in the control group treated with Captopril. The changes of symptom score, urinary microprotein series (urinary albumin excretion rate, Tamm-horsfall protein and beta 2-microglobulin), blood glucose, kidney function, blood lipid, angiotensin I (A I) and II (A II), atrial natriuretic polypeptide (ANP), thromboxane B2(TXB2), 6-keto-prostaglandin F1 alpha, endothelin 1 (ET-1) and collagen IV in patients after treatment were observed. RESULTS: The total effective rate in the TWKC group was 84.62%, which was superior to that in the control group (70.73%, P < 0.05). TWKC also showed better effects in improving clinical symptoms, lowering blood glucose, urinary microprotein series, blood lipid, A I, A II, ANP, ET-1 and collagen IV, ameliorating kidney function, and adjusting dynamic equilibrium of thromboxane-prostacyclin system, as compared with the control group (P < 0.05 or P < 0.01). CONCLUSION: TWKC could lower the levels of blood glucose and lipid, improve the glucose and lipid metabolism, regulate the microcirculation, ameliorate the degree of kidney damage, therefore, it showed a better effect in treating diabetic nephropathy. PMID- 11938825 TI - [Clinical study on effect of combined treatment with shenlong oral liquid and radiotherapy in treating nasopharyngeal carcinoma]. AB - OBJECTIVE: To observe the effect of Shenlong Oral Liquid (SLOL) combined with radiotherapy in treating nasopharyngeal carcinoma (NPC). METHODS: Effects of the combination therapy, including clinical effect, changes of cellular immunity and side effects, in treating 60 NPC patients (treated group) were observed and compared with those of the other 60 patients treated with radiotherapy alone (control group). RESULTS: (1) The side effect of radiation in the treated group was lower than that in the control group significantly (P < 0.05); (2) The immediate regression rate of nasopharyngeal and neck tumor in the two groups was not different significantly (P > 0.05); (3) The doses for complete remission of nasopharyngeal and neck tumor in the treated group was lower than that in the control group (P < 0.01); (4) No change of T-lymphocyte subsets was found in the treated group after treatment, but in the control group, OKT3, OKT4 and OKT4/OKT8 ratio were markedly decreased (P < 0.05); (5) The survival rate in the treated group was higher than that in the control group, but without statistical significance (P = 0.0518). CONCLUSION: The combination therapy of NPC by SLOL and radiotherapy is able to reduce side effect of radiotherapy, improve the cellular immunity, reduce the dose of radiation for tumor remission and enhance the therapeutic effect of radiotherapy, it might raise the survival rate of NPC patients. PMID- 11938824 TI - [Effect of different injections of Chinese herbal medicine on stress hormones and immune cell factors in patients of type 2 diabetes mellitus complicated with acute cerebral infarction]. AB - OBJECTIVE: To investigate the effect and clinical significance of different injections of Chinese herbal medicine on stress hormones and immune cell factors in treating patients of type 2 diabetes mellitus complicated with acute cerebral infarction. METHODS: Patients were divided into three groups, treated with Defibrase injection (DI, n = 32), Acanthopanax injection (AI, n = 20) and Ginaton injection (GI, n = 12) respectively. Parameters, including corticotropin releasing hormone (CRH), adrenocoticortropic hormone (ACTH), cortisol (CS), tumor necrosis factor alpha (TNF alpha) and interleukin-6 (IL-6) were determined using RIA in patients before and after treatment. The changes of parameters were analyzed and compared with those of healthy subjects for control. RESULTS: (1) Levels of all the above-mentioned parameters in all the three treated groups were higher than those in the healthy control group (P < 0.01); (2) All parameters were reduced after treatment in the three treated groups and the optimal effect was shown in the DI group (P < 0.01). CONCLUSION: The beneficial action of the three injections is closely related with the levels of stress hormones and immune cell factors, therefore, to monitor dynamically the changes of CRH, ACTH, CS, TNF alpha and IL-6 is of important significance in evaluation of therapeutic effect and elucidation of the pharmacology of Chinese herbal medicine. PMID- 11938826 TI - [Clinical observation on effect of yixin yidu decoction in treating acute viral myocarditis]. AB - OBJECTIVE: To observe the effect of Yixin Yidu Decoction (YXYDD, self-formulated according to the TCM therapeutic principle of supplementing Qi and Yin, clearing away evil Heat and toxic substances) in treating acute viral myocarditis (AVM). METHODS: Seventy-six patients of AVM were randomly divided into 2 groups, the 40 patients in the treated group were treated with YXYDD and the 36 patients in the control group treated with Qidong Yixin Oral Liquid (QDYX), the therapeutic course for both groups was 4 weeks. RESULTS: YXYDD was more effective than QDYX in improving clinical symptoms, adjusting the abnormal ECG, lowering the serum level of myocardial enzymes, improving the cardiac function of left ventricle and regulating the T-cell immunity (elevation of CD3, CD4 and CD4/CD8 ratio). CONCLUSION: YXYDD is an effective medicine in treating AVM. PMID- 11938827 TI - [Dynamic changes of serum pro-inflammatory cytokines and anti-inflammatory cytokines in patients with acute infection and the effect of Chinese herbal medicine intervention]. AB - OBJECTIVE: To study the dynamic changes of serum pro-inflammatory cytokines and anti-inflammatory cytokines in patients with acute infection and to explore the effect of 912 compound, a Chinese herbal preparation for intervention. METHODS: Forty patients with acute infection were randomly divided into 2 groups, 20 in the 912 group and 20 in the control group, and the levels of plasma endotoxin (lipopolysaccharide, LPS), serum pro-inflammatory cytokines, including tumor necrosis factor alpha (TNF alpha) and interleukin-6 (IL-6) and anti-inflammatory cytokines, including soluble TNF-RI, IL-4 and IL-10 were measured before and after treatment by using limulus reagent stroma azo-colored quantitative method and enzyme-linked immunosorbent assay (ELISA). RESULTS: The level of LPS increased at the early stage of acute infection, the levels of pro- and anti inflammatory cytokines were all elevated in various degrees, particularly obvious in IL-6, sTNF-RI, IL-4 and IL-10 and the elevations of the two were basically parallel. The TNF alpha, IL-6 and sTNF-RI were positively related with APACHE III scoring. The levels of LPS and inflammatory cytokines reduced to a different extent in accord with the improvement of condition after treatment, the levels of LPS, TNF alpha, IL-6 and IL-10 lowered more significantly in the 912 group than those in the control group (P < 0.05). CONCLUSIONS: LPS, pro-inflammatory and anti-inflammatory cytokines play important roles in the acute infection process, they were important indexes for evaluating severity of infectious diseases. The Chinese herbal preparation 912 compound could reduce the blood levels of LPS, TNF alpha, IL-6, IL-10 and prevent the decrease of sTNF-RI so as to promote the recovery of patients. PMID- 11938828 TI - [Clinical study on treatment of atherosclerosis obliterans by integrated traditional Chinese and Western medicine]. AB - OBJECTIVE: To investigate the mechanism in treating atherosclerosis obliterans (ASO) with integrated traditional Chinese and western medicine (TCM-WM). METHODS: The changes in level of vasoactive factors, such as endothelin (ET), nitric oxide (NO), apoprotein and the iconographic changes in angiography before and after treatment were observed in 40 patients of ASO. RESULTS: All patients had high levels of plasma ET and low level of NO, abnormal metabolism apoproteins, and showed the signs of trunk artery obstruction and marked decrease of collateral circulation in affected limb. After treatment, the plasma ET, NO and apoprotein levels recovered to normal range in 30 markedly effective cases, as compared with those before treatment, the difference was significant (P < 0.01) and their collateral circulation in the affected limb significantly increased, showing obvious improvement of peripheral circulation. CONCLUSION: The therapeutic mechanism of TCM-WM in treating ASO might be related with its effects on vascular endothelium, lipid metabolism adjustment, collateral circulation establishment and microcirculation improvement. PMID- 11938829 TI - [Clinical observation on treatment of pediatric intractable nephropathy with modified taohong siwu decoction]. AB - OBJECTIVE: To observe the clinical effect of Modified Taohong Siwu Decoction (MTSD) in treating pediatric intractable nephropathy (PIN). METHODS: Ninety-five cases of PIN were divided into 2 groups at random. The 60 cases in the treated group were administered with MTSD and the 35 patients in the control group were treated with heparin. The clinical therapeutic effect and levels of thromboxane B2(TXB2) and 6-keto-prostaglandin F1 alpha before and after treatment were observed. RESULTS: The total effective rate of the treated group was 81.7%, which was similar to that in the control group (80.0%, P > 0.05). The levels of TXB2 and TXB2/6-ketoprostaglandin F1 alpha ratio were higher in both groups of patients as compared with those of the healthy control (P < 0.01), After treatment, the two criteria were significantly improved in the two groups, as compared with those before treatment, the difference was significant (P < 0.01), while in comparson between the treated and the control group, no significant difference was found (P > 0.05). CONCLUSION: MTSD has good effect in treating PIN, it could improve the metabolic unbalance of thromboxane and prostacyclin. PMID- 11938830 TI - [Study on effect of spine surgery on gastric function and its efficacy of relevant treatments]. AB - OBJECTIVE: To study the effect of spine surgery (SpS) on gastric function and the efficacy of relevant treatments. METHODS: Sixty patients in the spine surgery group, 20 patients in the extremity surgery (ES) group and 20 healthy subjects in the control group were observed. Electrogastrography (EGG) was used to observe gastroelectric activity before and after operation. Twenty patients among the SpS group were examined with barium meal under actinoscopy to observe the gastric peristaltic waves before and after operation. The SpS group was randomly subdivided into 3 groups, and treated by Xiangsha Yangwei pill (XSYW), moxibustion and motilium respectively. At the same time, the EGG of various groups was observed and the change of preoperative and postoperative EGG were compared. RESULTS: The gastroelectric rhythm of SpS group was remarkably abnormal, both frequency and amplitude of EGG were significantly different from the other two groups (P < 0.05, P < 0.01). The gastric peristaltic waves were reduced, and the emptying time was obviously prolonged. EGG was significantly improved after either of the three treatments statistically (P < 0.05), the effect of XSYW was the best, but in comparing with the other two, the difference was not significant (P > 0.05). CONCLUSION: SpS could change the gastroelectric rhythm to cause the gastric functional disorder and induce gastroparesis, EGG can get satisfactory results in the diagnosis of these illnesses. XSYW, moxibustion and motilium all have significant effects on the gastric function after SpS. PMID- 11938831 TI - [Effect of kidney tonifying and circulation promoting compound on transcription of T-cell apoptosis related Fas and FasL genes in aged rats]. AB - OBJECTIVE: To probe the molecular mechanism for down-regulation of T-cell apoptosis by Kidney tonifying compound (KTC) in aged rats. METHODS: GAPDH was used as an internal control gene, and semi-quantitative analysis of the transcription of Fas and FasL genes was performed by using RNase protection assay. RESULTS: The Fas/GAPDH and FasL/GAPDH optical density (OD) ratios was 0.81 +/- 0.17 and 0.84 +/- 0.12 in the aged control group, and 0.36 +/- 0.16 and 0.26 +/- 0.12 in the young control group, the difference between the two groups was significant. In the circulation promoting group, the parameters were 0.99 +/- 0.36 and 0.94 +/- 0.33 which were not significantly different as compared with those in the aged control group (P > 0.05). In the two Kidney tonifying groups, the Fas/GAPDH OD ratio was 0.78 +/- 0.11 and 0.78 +/- 0.20, respectively, and no significant difference was observed as compared with those in the aged control group. However the FasL/GAPDH OD ratios in the two kidney tonifying group were 0.71 +/- 0.21 and 0.68 +/- 0.15, which were lower than those in the aged control group (P < 0.01). CONCLUSION: The negative regulation of FasL gene expression may be one of the molecular mechanisms of KTC in down-regulating excessive apoptosis of T-cell in aged rats. PMID- 11938832 TI - [Experimental study of protective effect of qingkailing on glutamate induced neurotoxic damage of brain]. AB - OBJECTIVE: To observe the effect of Qingkailing (QKL) on glutamate induced damage of brain, in order to seek for effective drugs for antagonizing neurotoxicity of glutamate. METHODS: Number and morphology metrology of neuron in cerebral cortex and hippocampus were detected by using MIAS-300 image analysing instrument, electron microscope and immunohistochemical methods. RESULTS: QKL could allevate the glutamate induced accumulation of water and sodium in the brain tissue, relief the damage of cerebral cells and reduce the percentage of c-fos positive cell in brain. CONCLUSION: QKL could protect brain damage induced by glutamate, which might be related to inhibition of QKL on glutamate caused enhancement of c fos gene expression. PMID- 11938833 TI - [Experimental study of daicong solution on brain M receptor in dementia model of aged rats]. AB - OBJECTIVE: To explore the effect of Daicong Solution (DCS) on M receptor in brain of aged rat model of dementia. METHODS: The effects of DCS on M receptor in brain of dementia model of learning and memory function hindrance were observed, the basal nuclei were ruined by destroying bilateral electrolytic method in 22-month old rats. RESULTS: Significant difference was shown in comparing the M receptor in the two groups treated by different dosages of DCS with that in the model control group (P < 0.01), significant difference also displayed in comparison of the DCS treated groups and Naofukang treated group (P < 0.01). CONCLUSION: DCS has marked effects in elevating the number and affinity of M receptor in brain of aged rat dementia model and could improve its learning and memory function, the effect was superior to that of Naofukang. PMID- 11938834 TI - [Effect of rhubarb on oxygen radicals leakage from mitochondria of intestinal mucosa in burned rats]. AB - OBJECTIVE: To investigate the effects of rhubarb on oxygen radicals leakage from the mitochondria of intestinal mucosa in burned rats. METHODS: The activity of cytochrome oxidase and superoxide dismutase (SOD) in mitochondria isolated from intestinal mucosa of burn model of rats was used to evaluate the oxygen radicals leakage from the mitochondria. And the SOD and malondialdehyde (MDA) levels in plasma, liver, intestinal mucosa and its mitochondria were also determined. RESULTS: After burn injury, the activity of cytochrome oxidase in intestinal mucosa decreased as the time elapses, rhubarb could improve it obviously. Furthermore, rhubarb could reduce the massive consumption of SOD and production of MDA obviously increased in plasma, liver, intestinal mucosa and its mitochondria after burning. CONCLUSION: Rhubarb could enhance the activity of cytochrome oxidase and SOD in small intestine mucosal epithelia in burned rats, thus the oxygen radicals leakage from the mitochondria to be reduced. PMID- 11938835 TI - [Experimental study on inhibitory effect of xiaochaihu decoction on duck hepatitis B virus]. AB - OBJECTIVE: To verify the therapeutic effect of Xiaochaihu decoction (XCHD) on chronic hepatitis B, and to prove the rationality of the TCM principle "strengthening the body resistance to eliminate pathogenic factors". METHODS: The inhibitory effects on DHBV of various doses of XCHD and its ingredients (grouped into whole recipe, partial recipe and single Bupleurum) were determined and compared, as well as compared with that of the blank control and of antiviral drug, acyclovir (ACV). RESULTS: Inhibitory effect was shown in all the treated groups. The treatment by whole recipe of XCHD with the dose 20 times that of clinical use showed the optimal effect, the difference in comparing with other treated groups was significant, P < 0.05. The effect of whole recipe was better than that of the partial recipe or single Bupleurum, P < 0.05. Moreover, the effect was rather persistent, no rebound phenomena was observed after withdrawal of medication, while in the ACV treated group, though a better effect conld be obeained, however, the DHBV returned to the level of before treatment after cessation of ACV treatment. CONCLUSION: The therapeutic effect of XCHD might be to supplment body resistance and remove the evil pathogen, strengthen or regular immune function, so XCHD is efficient in treating chronic hepatitis B. PMID- 11938836 TI - [Experiences of Doctor Ye Shaoli in the treatment of children's diseases]. PMID- 11938837 TI - [Traditional Chinese medicine therapy of secondly hypoxia pulmonary arterial hypertension caused by chronic obstructive pulmonary diseases]. PMID- 11938838 TI - [Summary of the national meeting on speech pathology and speech rehabilitation]. PMID- 11938840 TI - [A probe into the relationship between spasmodic dysphonia and laryngeal paralysis]. AB - OBJECTIVE: To investigate into the relationship between spasmodic dysphonia and laryngeal paralysis. METHODS: The intrinsic laryngeal muscle potential was recorded with electromyography. Vocal cord movements were observed with a videostroboscope. Laryngeal paralysis was divided into mild, moderate, and severe degrees based on the potentials of intrinsic laryngeal muscles and on the status of vocal cord movements. RESULTS: In the past 12 years (from 1983 to 1994) 1300 cases were diagnosed as having mild, moderate, and severe laryngeal paralysis. Among them, there were 5 cases with laryngospasm including 3 mild, 1 moderate, and 1 severe case. CONCLUSION: The findings obtained from careful observation on these 5 cases of spasmodic dysphonia demonstrated that there existed certain relationships between laryngeal paralysis and spasmodic dysphonia. During the course of exacerbation or restoration of paralysis, spasmodic dysphonia might occur. PMID- 11938839 TI - [Visceral voice training for laryngectomy after hypopharyngectomy and visceral transplant]. AB - OBJECTIVE: To establish a way to visceral voice after stomach or colon transplantation for patients with hypopharyngeal or cervical esophageal cancer. METHODS: Esophageal voice training was used and modified according to the compliance of the patients. Twenty laryngectomies with replacement of the hypopharynx and esophagus by stomach(13 cases), colon(6 cases) and jejunum (1 case) were trained for voice rehabilitation at the Department of Head and Neck Surgery. RESULTS: The patients with an artificial esophagus from vicera were easy to gain an influx of certain volume of gas into their artificial esophagus (stomach or colon) and to learn to speak. But on the whole the quality of voice was not so satisfactory. In this series nineteen out of 20 patients (95%) could express their idea by speech after a training course of three weeks. CONCLUSION: The literature has emphasized role of cricopharyngeus muscle in the training of esophageal voice. Owing to the fact that this series of patients who had had their cricopharyngeus removed, could easily get their voice rehabilitated, it seems this muscle played no major role in the voice rehabilitation. PMID- 11938841 TI - [Experimental study on human collagen and autologous fat injected into canine vocal cords]. AB - OBJECTIVE: To explore the histological fate of human collagen and autologous fat injected into vocal cords in treating glottic insufficiency. METHODS: Human collagen and autologous fat were injected respectively into two dogs(group A) to correct glottic incompetence caused by left vocal cord paralysis. Autologous fat and human collagen 0.3 ml were injected respectively into left and right vocal cords of 9 dogs (B, C, D groups, 3 dogs in each group). These animals were killed after 1, 3 and 5 months. The histology, volume and curative effect of both substances were evaluated. RESULTS: Autologous fat grafts survived in the canine vocal cords after 5 months but the absorption was rapid. The average speed of reduction in volume was 38.51% from 1 to 5 months and the effect of treatment couldn't last long. Human collagen was easily injected and well-tolerated. The injected collagen was colonized by host fibroblasts and capillaries. At the same time, new host collagen deposited. There was a trend to allow the eventual replacement of the implant by new host tissue. The average speed of reduction in volume was 0.65% and the curative effect was constantly stable from 1 to 5 months. CONCLUSION: The results suggested that autologous fat has only a potential use for short-term(< 5 months) augmentation of vocal cord. Human collagen can be used as long-term augmentative material to manage glottic insufficiency caused by various etiologies. PMID- 11938842 TI - [The influence of changes in vocal cords' cover-body on their vibratory pattern]. AB - OBJECTIVE: To discuss the influence of changes in vocal cords' cover-body on their vibratory pattern. METHODS: Fifty adult patients with vocal polyps were observed by telescopic videolaryngostroboscopy and detected by electroglottography (EGG). We examined the vocal cords' vibratory pattern according to laryngostroboscopic image and discussed the change of vocal cords' vibratory parameters and EGG curves. RESULTS: The vocal cords' vibratory parameters and EGG curves changed with different vocal polyps. All vocal polyps affect the regularity(REG), amplitude (AMP), glottal closure(GLO) and mucosal wave(MUC) of laryngostroboscopic image; the symmetry(SYM), REG and GLO bore linear correlation to the notches in EGG curves, the REG and MUC bore linear correlation to the steep changes; micro-polyps with pedicles reduced the vibratory parameters CQ and F0, the main abnormalities were notches in EGG curves; the changes in macro-polyps with pedicles were the same as the micropolyps except increased CQ. All vocal polyps with wide bases either single or bilateral, had increased CQ and F0, the main abnormalities were steep in contact opening phase and flat in wave peak. CONCLUSIONS: Studies with laryngostroboscopy and EGG revealed how the vocal polyps affected vocal cords' cover-body, changed the vibratory pattern of vocal cords, and factors regulating vocal cords' vibratory pattern. PMID- 11938843 TI - [Changes in endoneurial fluid pressure in the crushed facial nerve]. AB - OBJECTIVE: To investigate the changes in endoneurial fluid pressure (EFP) and diameter of the crushed facial nerves in guinea pigs in order to determine whether there is necessity and the opportunity of the facial nerves decompression in different stage after injury. METHODS: EFP were measured using a servo-nulling micropipette system in the normal and crushed facial nerves. RESULTS: The results showed that the EFP in the facial nerve increased significantly after compression. The highest pressure of EFP 12.94 +/- 1.69 mmHg occurred at the end of the first week after injury, which was 4 to 5 times as great as that of normal ones. This change lasted about two weeks, then it returned to normal at the third or the fourth week. The largest diameter of the facial nerve 1.01 +/- 0.08 mm was seen at the end of the first week after crush injury. CONCLUSION: It suggested that the servonulling micropiette system was useful in evaluating the pathogenesis of facial palsy. It could provide the laboratory basis for facial nerve decompression. PMID- 11938844 TI - [Histopathologic study on intratemporal minimal facial nerve neurilemmomas]. AB - OBJECTIVE: To explore the original location and pathologic behavior of facial nerve neurilemmomas. METHODS: Human temporal bone slides from 815(1,526 ears) were histopathologically observed under light microscope. RESULTS: Seventeen(1.84%) undiagnosed intratemporal facial nerve neurilemmoma with morphologic indications of Antoni type A were found. According to pathologic features, the intratemporal facial nerve neurilemmomas were classified as ingrowing facial nerve neurilemmomas (4 ears) which originated from the center of facial nerve, and outgrowing facial nerve neurilemmomas (13 ears) which originated from the perineurium at dehiscence of the facial nerve canal. Of the 17 tumors, two originated from the labyrinthine segment, twelve from the tympanic segment, one from second turn and two from vertical segment. CONCLUSION: It is of important pathologic and clinical significance to classify the intratemporal facial nerve neurilemmomas into outgrowing and ingrowing types. PMID- 11938846 TI - [The normal value and clinical applications of the blink reflex]. AB - OBJECTIVE: To study the normal value and clinical applications of blink reflex. METHODS: Using the clinical electrodiagnostic techniques, the orbital nerve was stimulated to elicit the blink reflex. RESULTS: In 20(40 sides) normal subjects, two components (R1 and R2) had been documented. The latencies were about 10.1 ms for R1, 30.4 ms and 30.7 ms separately for R2 and R2'. In 30 patients with trigeminal and facial nerve diseases and acoustic neuromas, the latencies on the affected sides were apparently longer or the reflex disappeared (P < 0.01). CONCLUSION: This study indicates that the simple, harmless electrodiagnostic measurements are of value in the initial investigation of trigeminal and facial nerve diseases and acoustic neuroma. PMID- 11938845 TI - [Microvascular decompression of pons for idiopathic hemifacial spasm and exploration of its mechanism]. AB - OBJECTIVE: To further explore the cause, mechanism and surgical treatment of idiopathic hemifacial spasm (IHFS). METHODS: Routine sagittal and coronal cryoslices of 8 (16 sides) pons were made. The sections were stained with Luxol fast blue and Nissl stains. The location of the facial nucleus and pathway of facial neurofibers in the pons were observed. Twenty-five patients with IHFS whose facial nerve root exit zone(REZ) was not compressed by vessels underwent microvascular decompression of pons. RESULTS: The distances from the facial nucleus to the ventral surface of pons, the median line, and the ventroexterior surface were 12.0-12.5 mm, 6.0 mm, and 7.5-9.0 mm respectively. The distance from the facial neurofibers in the pons to the surface of the pons was 1.0 to 2.0 mm, with an average of 1.65 mm. The vascular compression was at the surface of pons, within 4 mm anterosuperior to the REZ in 25 IHFS patients. The compressing vessels were the anterior inferior cerebellar arteries and their branches. The results of decompression in 25 patients with IHFS were as follows: spasm immediately disappeared in 8 patients, and within one week in 17 patients. No recurrence was found during a follow-up period from 6 months to 5 years. CONCLUSION: The main cause of IHFS was vascular compression, but the position of compression is not necessarily always at the REZ. The "short-circuiting" theory can only explain the cases with vascular compression at the REZ. But in cases who do not have vascular compression at the REZ, there might be dysfunction of the facial nucleus. In the present cases, vascular compression was at the surface of the brain stem anterosuperior to the REZ, pons microvascular decompression is an effective treatment. PMID- 11938847 TI - [Expression of intercellular adhesion molecules in nasal polyps tissue and its significance]. AB - OBJECTIVE: To study the expression of intercellular adhesion molecule 1/lymphocyte function-associated antigen-1 in nasal polyps tissue and its significance. METHODS: Frozen sections of 9 normal nasal mucosa and 19 nasal polyp samples were studied with immunohistochemical technique and immunohistochemical and MGG dual staining method with ICAM-1 and LFA-1 monoclone antibodies. RESULTS: Expression of ICAM-1 and LFA-1 were stronger in nasal polyps tissue than in the normal nasal mucosa, and there is positive relationship between the expression of ICAM-1 and the expression of LFA-1 on eosinophils. CONCLUSION: High expression of ICAM-1 and LFA-1 in nasal polyps tissue showed that interaction of ICAM-1/LFA-1 may participate in the inflammatory reaction and prompt adhesion and migration of eosinophils. High expression of LFA-1+ on eosinophils may play an important role in the pathogens of nasal polyps. PMID- 11938848 TI - [The clinical application of functional endoscopic sinus surgery and conventional technique in sinus surgery]. AB - OBJECTIVE: To properly understand the basic theory and the clinical application of the functional endoscopic sinus surgery and the traditional technique. METHODS: The indications, extend of excision, operative approach, complication and curative effect of 1,740 cases(3,140 sides) done in the China-Japan Friendship Hospital and Jining Jiaotong Hospital were analysed. Among them, 522 cases(1,032 sides, 30%) received the traditional technique for sinusitis; 1,218 cases(2,108 sides, 70%) received functional endoscopic sinus surgery. RESULTS: Three hundred and ninety-eight cases of traditional ethmoid sinus operation and 352 of functional sinus surgeries were followed-up and their cure rates were 80% and 93% respectively. Both groups had no serious complications. CONCLUSIONS: The main indications of functional sinus surgery are infectious nasal sinus diseases or meatus nasi lesion that may interfere with the drainage. Such kinds of diseases have high morbidity than the diffused polyposis and deserves attention. Traditional operations for frontal and maxillary sinusitis should be controlled appropriately, but the traditional sphenoid sinus operation is still an effective treatment for the diffused polyposis. PMID- 11938849 TI - [Application of maxillary bone eversion reduction on benign tumor in nasal cavity and paranasal sinus invading the skull base]. AB - OBJECTIVE: To investigate the operative methods for benign tumors in nasal cavity and paranasal sinus invading the skull base. METHODS: Six cases of benign tumors in nasal cavity and paranasal sinus invading the skull base were reported. Of them, 3 were ossifying fibromas, 1 osteoma, 1 osteochondroma and 1 giant osteoid osteoma. The tumors were removed by combined approaches through unilateral maxillary bone eversion reduction. RESULTS: No complication was found. Follow-up was made 2-3 years after operation. Five cases were uneventful, 1 had a relapse of tumor but still alive. CONCLUSIONS: The method is appropriate for treating benign tumors in nasal cavity and paranasal sinus invading the skull base. PMID- 11938850 TI - [Endoscopic nasal surgery in treatment of nasopharyngeal angiofibroma]. AB - OBJECTIVE: To avoid complications of the routine surgeries for nasopharyngeal angiofibroma. METHODS: Endoscopic nasal surgery and other new techniques including controlled-hypotension anesthesia and preoperative feeding artery embolism had been used. From April 1996 to September 1997 eight young male patients with diagnosis of nasopharyngeal angiofibroma were treated by endoscopic sinus surgery. The average age was 16.6 years old. Six accepted preoperative embolism, seven accepted hypotension anesthesia. RESULTS: No complications had occurred. Average intraoperative blood loss was 900 ml. All cases were followed for 1 to 20 months(mean 8 months). Recurrence was found in only one patient three months after operation. The recurrent rate was 12.5%. CONCLUSION: The lesions limited to nasal and nasopharyngeal cavities and with sphnoid and ethmoid invasions can be removed by endoscopic nasal surgery. Endoscopic surgery has advantages in reducing complications and obtaining better post operative functions. Transnasal endoscopic examination is also necessary in the follow-up period. Other techniques such as CT, digital subtract angiography, preoperative feeding artery embolism and controlled-hypotension technique during anesthesia are beneficial. PMID- 11938851 TI - [DNA content, expression of proliferating cell nuclear antigen and surgical margins in locally advanced laryngeal cancer]. AB - OBJECTIVE: To probe into the relationship between DNA content, PCNA and prognosis. METHODS: Surgical margins in 34 patients with locally advanced laryngeal cancers were observed by means of antibody to PCNA and DNA content measurements. RESULTS: The results demonstrated that at neoplasm margins of 1.0, 0.5, and 0 cm, PCNA indices were 8.62%, 17.76% and 50.32% and DNA indices were 0.98, 1.082 and 1.436 respectively. There is a significant difference between 1.0, 0.5 and 0 cm. CONCLUSION: Moderate and severe atypical hyperplasia of epithelium (AHE) was different from mild AHE and simple hyperplasia of epithelium in the biologic behavior of fission, proliferation and so on and surgical margins should be kept outside the area of moderate AHE. PMID- 11938852 TI - [Preoperative radiation plus surgery vs. operation alone for laryngeal carcinoma]. AB - OBJECTIVE: To study the validity of preoperative radiation in improving the survival rate of laryngocarcinoma. METHODS: A prospective randomized clinical trial was conducted with preoperative radiation plus surgery (RS) as one group and surgery alone (SA) as the other group. The ages of the patients were less than 75 years, and the operative types were decided after clinical examination. The patients were divided into SA or RS group by random. Doses of preoperative irradiation was 40 Gy. Three hundred and seventy patients with laryngocarcinoma were treated, 215 in the SA group; 155 in the RS group. All patients were followed up over three years. RESULTS: In SA group, 3 years survival rate was 83.3%, 5-year survival rate 82.6%, 10-year survival rate 80.3%. In RS group, 3 year survival rate was 78.9%, 5-year survival rate 76.4%, 10-year survival rate 68.6%. There is no statistically significant difference in survival rates between the groups (P = 0.1); but, the 10-year survival rate in SA group was better than that in RS group for the stage III and IV supraglottic carcinomas, the former was 73.6% and the latter 63.5% (P = 0.003). The 10-year survival rate for the patients with T3 and T4 supraglottic carcinomas who underwent total laryngectomy in SA group was better than that in RS group. The former was 68% and the latter 50% (P = 0.0001). CONCLUSION: Preoperative radiotherapy with 40 Gy didn't increase the survival rate of laryngeal carcinoma. The 10-year survival of stages III and IV supraglottic carcinoma in combined treatment group was lower than that in the surgery alone group. PMID- 11938853 TI - [A preliminary study on the relationship between nutritional status and postoperative complications in patients with head and neck malignancies]. AB - OBJECTIVES: To study the role of nutritional factor in the development of postoperative complications in patients with head and neck malignancies. METHODS: Postoperative complications of 96 patients with head and neck malignancies who were operated on were surveyed. Twenty-seven developed postoperative complications; the remaining 69 went uneventful. Comparisons of nutritional status and clinical variables between these two groups were made. RESULTS: 1. The prevalence of poor preoperative nutritional status in the complication group (56%) was higher as compared with that in the noncomplication group (20%) (P < 0.001); 2. values of body weight, triceps skin fold thickness, arm circumference, arm muscle circumference, and creatinine-height index in the complication group decreased much more significantly than in the noncomplication group (P < 0.001); 3. the nitrogen and calorie intake within the first postoperative week was lower in the complication group than in the non-complication group (P < 0.001). CONCLUSIONS: Nutritional factor plays a very important role in the development of postoperative complication and the perioperative nutritional support for head and neck surgical patients should not be ignored. PMID- 11938854 TI - [Mandibular swing procedure for resection of pharyngeal and skull base tumors]. AB - OBJECTIVE: To seek for a better approach for the resection of pharyngeal and skull base tumors. METHODS: The overall tumor distribution for the entire group was nasopharyngeal 2 cases, oropharyngeal 4, parapharyngeal space 5 and parapharyngeal infratemporal area 2 cases. All 13 cases of pharyngeal and skull base tumors had their tumors thoroughly resected via mandibular swing approaches. Five cases had retropharyngeal node dissections, 2 cases had unilateral modified radical neck dissections, 5 cases with defect oropharynx were reconstructed with the pectoralis major myocutaneous flap, 10 cases with malignant tumors had received adjutant radiotherapy after surgical procedures. RESULTS: Of the 13 cases, 10 were malignant, 3 were benign. The incision in 12 cases healed primarily, one case with malignant fibrocystic tumor got infective necrosis of pectoralis major myocutaneous flap, this case healed completely after more than two months. One case developed dysphagia postoperatively and was recovered by swallow training. One case had minimal occlusion disorder. All patients were followed up from 15 months to 3 years. Three cases with benign tumors achieved good clinical results and are living well. In the malignant group, one died of recurrence at 6 months postoperatively, one died of lung metastasis, 2 cases survived for 3 years, 2 for two years, 4 for one year. CONCLUSION: This procedure provides good exposure of the base of the skull, the pharynx and the parapharyngeal space as well as the clivus and upper cervical vertebrae; it allows dissection along the internal carotid artery and facilitates resection of the tumor en bloc. It provides operative safety and minimal morbidity. PMID- 11938855 TI - [Diagnosis and treatment of systemic lupus erythematosus with integrated traditional Chinese and Western medicine]. PMID- 11938856 TI - [Study on improvement of liver cirrhosis and liver function in hepatolenticular degeneration patients treated with integrated Traditional and Western medicine]. AB - OBJECTIVE: To observe the effects of sodium dimercaptosulphonate (DMPS) plus Gandou tablet, DMPS and calcium disodium ethylene diaminotetraacetate (EDTA) on improving liver cirrhosis and liver function of hepatolenticular degeneration (HLD) patients. METHODS: One hundred and forty-six HLD patients were divided into A, B, C three groups, and treated with DMPS plus Gandou tablet, DMPS and EDTA respectively, the therapeutic course was 8 weeks for three groups. The ultrasonography of liver, electrophoresis of serum protein and excretion of urinary copper were observed. RESULTS: The ultrasonography of liver was improved in all groups, the rate of improvement of group A was 54.0%, B was 44.0% and C was 39.1%. The amounts of serum albumin in A and B groups increased (P < 0.01, P < 0.05), and gamma-globulin decreased in all groups (P < 0.05). The excretion of urinary copper increased obviously in all groups (P < 0.01), and A, B groups increased more than that of C group (P < 0.05). CONCLUSIONS: The de-copper therapy could improve liver cirrhosis and liver function. The effect of DMPS plus Gandou tablet was better than that of DMPS and EDTA. PMID- 11938857 TI - [Clinical study on treatment of infantile rotaviral enteritis with Psidium guajava L]. AB - OBJECTIVE: To observe the clinical effect of olium Psidium guajava (PG) in treating infantile rotaviral enteritis. METHODS: Sixty-two patients of rotaviral enteritis were randomly divided into the treated group treated with PG and the control group treated with Gegen Qinlian decoction. The time for ceasing diarrhea, the content of Na+ in blood, the content of Na+ and glucose in stool, and the rate of negative conversion of human rotavirus (HRV) antigen were observed. RESULTS: The rate of recovery in 3 days of the treated group was 87.1%, significantly higher than that of the control group (58.1%, P < 0.05). The time of ceasing diarrhea of the treated group (25.1 +/- 9.5 hr) was significantly shorter than that of the control group (38.7 +/- 15.2 hr, P < 0.01). The content of Na+ and glucose in stool was reduced obviously in the treated group, while the reduction in the control group was insignificant, the treated group was superior to the control group significantly (P < 0.05). The rate of negative conversion of HRV in feces of the treated group was 87.1%, significantly better than that of the control (58.1%, P < 0.05). CONCLUSION: The treatment with PG has good curative effect on infantile rotaviral enteritis. PMID- 11938858 TI - [Effects of Salvia miltiorrhiza compound injection on serum endothelin, prostaglandin I2/thromboxane A2 ratio alteration following myocardial ischemia reperfusion in patients undergoing intracardiac surgery]. AB - OBJECTIVE: To investigate the effects of Salvia miltiorrhiza compound injection (SMCI) on serum endothelin (ET), prostaglandin I2(PGI2), thromboxane A2(TXA2), and PGI2/TXA2 ratio following myocardial ischemia-reperfusion in patients undergoing intracardiac surgery. METHODS: Twenty patients, scheduled for selective surgery, were randomly divided into the SMCI group (group A, 10 cases) and the control group (group B, 10 cases). SMCI 200 mg/kg was given intravenously in group A before starting the operation and at the time of rewarming respectively, and equivalent volumes of normal saline were administered to group B. The central venous blood samples were collected to measure the serum concentration of ET, PGI2, TXA2, and PGI2/TXA2 ratio. RESULTS: ET significantly reduced, while PGI2 and TXA2 obviously raised in both groups at the beginning (T1) of extracorporeal cardiopulmonary bypass (CPB) (P < 0.05). After cardiac ischemia-reperfusion, ET in group B increased rapidly and significantly (P < 0.05) and evidently higher than the corresponding value in group A 30 min after reperfusion (T3) till 24 hrs after reperfusion (T5). During reperfusion, PGI2 and TXA2 in group A decreased more rapidly than that of group B, while group A maintained higher PGI2/TXA2 ratio than that of group B. At T5, PGI2 and TXA2 in group A were significantly lower than those in group B, while PGI2/TXA2 ratio was higher than that in group B (P < 0.05). The serum ET level was obviously negatively correlated to PGI2/TXA2 ratio. The postoperative cardiac function recovered much better in group A than in that group B. CONCLUSION: SMCI can significantly reduce serum ET level, raise PGI2/TXA2 ratio, thus facilitate the postoperative cardiac function recovery following intracardiac surgery under CPB. PMID- 11938859 TI - [Study on lumbricus in promoting ligation hemorrhoidectomy postoperative wound healing]. AB - OBJECTIVE: To investigate the effect of Lumbricus in promoting wound healing of ligation hemorrhoidectomy. METHODS: Spray the artificial grown fresh Lumbricus solution on the wound surface of the mixed hemorrhoid patients (treated group) after hemorrhoidectomy and also on wound of the experimental animals, and Lithospermum erythrorhizon medicated gauze was taken as control to observe the effect of treatment on wound healing and histologic change. RESULTS: The mean wound healing time of the treated group was 16.5 +/- 1.8 days, as compared to the control group (21.2 +/- 2.8 days), it was obviously shortened (P < 0.01), 3 days after medication, the growth of epidermis of the treated group was obviously more rapid than that of the control group, no wound infection and granulation hyperplasia were found. The experimental study showed that Lumbricus preparation could inhibit inflammation, bacteria such as Staphylococcus aureus, coli and proteus bacillus was inhibited. In comparison with the control group, the experimental group shortened the healing period for 4 days. On the 4th and 7th day, sacrificed rabbits' capillary count, vessel lumen endothelial cell count, and mesenchymal fibroblast count of the experimental group grower obviously than those of the control group, on the 4th day, the mesenchymal cell mitosis of the experimental group was higher than that of the control group, while on the 7th day, the cell nucleus mitotic index in the experimental group also was higher than that in the control group, from the 3rd day on, wound healing and granulation filling speed of the experimental group obviously more rapid than that of the control group. CONCLUSION: The Lumbricus preparation is cheap in price, easy to preserve, can be used in promoting wound healing, without any toxic and side-effects. PMID- 11938860 TI - [Effect of yinchen dandao decoction on plasma fibronectin, biliary thromboxane A2 and prostaglandin I2 in human and animal suffering from acute cholangitis]. AB - OBJECTIVE: To study the therapeutic mechanism of Yinchen Dandao decoction (YCDDD) in treating acute cholangitis. METHODS: Twenty-four patients with acute cholangitis and 36 cholangiolithiasis patients were randomly classified into the treated group and the control group, and all of these patients had undergone surgical operation. The treated group were given orally YCDDD 3 days after operation for 1 week, and plasma fibronectin (FN), biliary thromboxane A2(TXA2) and prostaglandin I2(PGI2) were measured and compared before and after oral administration of the drug. The models of cholangitis were established in rabbits, which were also randomly grouped into the treated group and the control group, and the same parameters (FN, TXA2, PGI2) were observed as in rabbits. RESULTS: The level of FN was significantly lower in cholangitis patients than that in the cholangiolithiasis patients before operation (P < 0.01), and compared to the control group, it obviously raised after YCDDD treatment (P < 0.01), the same happened in the cholangiolithiasis patients. The value of TXA2 and PGI2 in the bile was obviously higher (P < 0.01, P < 0.05) in cholangitis patients than those in chlangiolithiasis, and obviously reduced after YCDDD administration (P < 0.01). The level of FN in the treated group and the level of biliary TXA2 and PGI2 in rabbits with cholangitis were significantly higher than those in the control group (P < 0.01). CONCLUSION: YCDDD caused the increase of FN and decrease of biliary TXA2 and PGI2, which might be one of the mechanisms of the drug carried out in cholangitis patients. PMID- 11938861 TI - [Study on relationship between estrogen receptor gene polymorphism and syndrome differentiation typing of female postmenopausal osteoporosis in Traditional Chinese medicine]. AB - OBJECTIVE: To study the relationship between estrogen gene polymorphism and TCM Syndrome Differentiation of female postmenopausal osteoporosis in China. METHODS: Two hundred and forty-six Chinese postmenopausal women, age 44-80 years, mean 65.8 years, using molecular biological method to analyze the endonuclease Pvu II, Xba I restriction fragment length polymorphisms (RFLPs), with dual X-ray bone mineral density absorption meter to determine the bone mineral densities of lumbar vertebra (L1-4) and femur (intertrochanter, femur neck, Ward's region) separately. The subjects were divided into Kidney Yin deficiency type, Kidney Yang deficiency type and both Kidney Yin-Yang deficiency type, to observe the relationship between TCM and bone density as well as estrogen receptor gene polymorphism, Pp(Pvu II) and Xx(Xba I) were used to express RFLPs, the capital P and X to express the deficit of restricting sites. RESULTS: Bone mineral density of PPxx gene type (n = 21) was obviously lower than that of other gene types (n = 225), lumbar (-0.71 +/- 0.46) g/cm2, intertrochanter (-0.31 +/- 0.58) g/cm2, femur neck (-0.84 +/- 0.66) g/cm2, Ward's region (-0.96 +/- 0.85) g/cm2, the TCM Syndrome Differentiation typing of this gene type belonged to both Kidney Yin Yang deficiency type. CONCLUSION: Estrogen receptor gene RFLPs is related to TCM Syndrome Differentiation typing. PMID- 11938862 TI - [Exploration on parameters of TCM syndrome in acute cerebral infarction through investigating active factors of vascular endothelium cells]. AB - OBJECTIVE: To explore the multiple parameters of TCM Syndrome-types and the acute cerebral infarction (ACI) with blood stasis type. METHODS: Sixty-six acute cerebral infarction patients with blood stasis Syndrome, various vascular active factors such as tissue plasminogen activator (t-PA), the activity of plasminogen activator inhibitor (PAI), the concentration of prostaglandin F1 alpha (6-keto PGF1 alpha) etc. were determined. RESULTS: (1) In Incidental Syndrome, those "Phlegm" and "stasis" predominant, mainly manifested as Wind-Phlegm-Blood stasis (WPBS), Qi deficiency-blood stasis (QDBS) and Phlegm-Heat-bowel excess (PHBE) Syndrome all showed t-PA activity lowered, among them, QDBS Syndrome lowered more obviously (P < 0.01); and in fundamental deficiency predominant Syndrome such as Yin-deficiency and Wind-move (YDWM) Syndrome, the active t-PA content increased (P < 0.05); in Liver Yang ascending (LYA) Syndrome and YDWM Syndrome, the 6-keto PGF1 alpha lowered very significantly. (2) Through regression analysis, although influencing the severity of acute blood stasis was related with 3 factors (t-PA activity, nervous system defect score and age growth), but single factor linear relationship analysis indicated that did not existed positive-negative relationship. (3) Through statistical analysis, the factor influencing nervous system defect scores was positively related with blood stasis score (r = 0.70, P < 0.01). CONCLUSION: (1) The basis of WPBS, QDBS and PHBE Syndrome mainly was fibrinolytic system activity lowering, and YDWM and LYA Syndrome prostaglandin system activity lowering. Comprehensive analysis of multiple parameters would be helpful to differentiate the ACI blood stasis stage. (2) Single parameter would not help to differentiate the types of ACI blood stasis stage, its change merely denoted the existence of blood stasis, its type should be differentiated after comprehensive analysis. (3) Those influencing nervous system scoring, mainly was blood stasis score, which suggested that the importance of activating blood circulation to remove stasis in preventing and treating ACI. (4) Put forward ACI blood stasis, and the quantification for new standard of Syndrome for discussion. PMID- 11938863 TI - [Treatment of hypertension by jiangya oral liquid]. PMID- 11938864 TI - [Experimental study on suppressive effect of ginkgo extract on adhesion of vascular endothelial cell to monocyte induced by minimally modified low density lipoprotein]. AB - OBJECTIVE: To observe the effect of Ginkgo extract on the interaction between minimally modified low density lipoprotein (MM-LDL) induced human umbilical vein endothelial cell (HUVEC) and U937 monocyte-like cell line. METHODS: The adhesive percentage between HUVEC and U937 by counting and the expression of VCAM-1, ICAM 1, P-selectin by ELISA. RESULTS: Treatment of HUVEC with MM-LDL (75 micrograms/ml) for 4 hrs could significantly increase the adhesion of U937 to HUVEC (P < 0.01) and Ginkgo extract could suppress the adhesion in dose-dependent manner. The surface expression of VCAM-1, ICAM-1, P-selectin was not induced by MM-LDL, while recombination tumor necrosis factor-alpha 5.0 ng/ml, as a positive control, could do. CONCLUSION: Ginkgo extract afforded protection against HUVEC damage induced by MM-LDL and the adhesion of monocytes to endothelial cells induced by MM-LDL is not mediated by VCAM-1, ICAM-1 and P-selectin. PMID- 11938865 TI - [Effects of yanggan xifeng recipe on changes of neurological behavior of rat of Parkinson's model]. AB - OBJECTIVE: To investigate the effect of Yanggan Xifeng Recipe (YGXFR) for nourishing Liver to calm down endogenous Wind on neurological behavior of Parkinson's disease (PD) rat model. METHODS: PD rat model was established by destroying the substantia nigra condense area (SNc) and ventral tegmental area (VTA) unilaterally with 6-hydroxy dopamine (6-OHDA). The rats for control were injected in the same site with the same dose of normal saline. The animals were randomly divided into 3 groups (7 in each group), the control group, the model group and the TCM treated group. Staircase test and rotation test induced by apomorphine were performed in all groups and compared. RESULTS: The staircase test demonstrated that, after being treated with YGXFR, the food pellets intake were increased compared to model group (P < 0.05), and their rotation induced by apomorphine was lessened (P < 0.01). CONCLUSION: YGXFR could recover the neurological behavior deficits of PD rat models. PMID- 11938866 TI - [Study on mechanism of Shenle and Fragmin in preventing and treating glomerulosclerosis in rats]. AB - OBJECTIVE: To study the effects of Shenle and Fragmin on the degradation of extracellular matrix metalloproteinase-9 (MMP-9) and tissue inhibitors of metalloproteinase-1 (TIMP-1) in the remnant kidneys of 5/6 nephrectomized rats treated with Shenle and Fragmin. METHODS: The model of focal and segmental glomerulosclerosis was made by 5/6 nephrectomy in Wistar rats. After being treated with Shenle (4 g.kg-1.d-1) or Fragmin (1000 IU.kg-1.d-1) for 20 weeks, the blood pressure, proteinuria, serum creatinine, lipids, renal pathological change of remnant kidney of the model animals were examined, and the mRNA expression of MMP-9, TIMP-1 and type IV collagen in the remnant kidney were also detected. RESULTS: Shenle and Fragmin could significantly reduce the blood pressure, proteinuria, serum creatinine and lipids, alleviate the pathologic change of kidney tissue, decrease the type IV collagen, MMP-9 deposition. Shenle and Fragmin down-regulated the mRNA expression of MMP-9 by 30.0% and 28.5% and TIMP-1 expression by 59.3% and 55.0% in the remnant kidney of the 5/6 nephrectomized rats respectively. CONCLUSION: Shenle and Fragmin might ameliorate glomerulosclerosis through modulating MMP-9/TIMP-1 gene expression, alleviate the abnormal metabolism and accumulation, promote extracellular matrix degradation in glomeruli remodeling, it might be the important mechanism of Shenle and Fragmin in treating glomerulosclerosis. PMID- 11938867 TI - [Apoptosis of human gastric cancer cell line MGC-803 induced by glycyrrhiza uralensis extract]. AB - OBJECTIVE: To study the apoptosis in human gastric cancer cell MGC-803 induced by the extract of glycyrrhiza uralensis Fisch. METHODS: Apoptosis was detected by laser scanning confocal microscope, agarose gel electrophoresis and flow cytometry. The p53 gene expression was analyzed by double immunolabelling and immunohistochemistry technique. RESULTS: Chromatin condensation of different phases of apoptotic cells were observed by laser scanning confocal microscope, the percentage of apoptosis determined by flow cytometry was time- and dose dependent, agarose gel electrophoresis of DNA exhibited obvious "ladder" in the cells treated with higher concentration of the extract of glycyrrhiza uralensis. The p53 gene expression was unchanged after treatment. CONCLUSION: Glycyrrhiza can induce human gastric cancer cell line MGC-803 to apoptosis by p53-independent pathway and can be used as a potential, natural apoptosis-inducing agent for gastric cancer therapy. PMID- 11938869 TI - [Primarily study on clinical quality analysis and therapeutic economics]. PMID- 11938868 TI - [Effect of shenkang injection on transforming growth factor-beta messenger ribonucleic acid of LLC-PK1 renal tubular epithelial cells]. AB - OBJECTIVE: To study the effect of Shenkang injection (SKI) on the transforming growth factor (TGF)-beta mRNA of LLC-PK1 renal tubular epithelial cells. METHODS: Using reverse transcriptase-polymerase chain reaction (RT-PCR) and Northern Blot to observe the effect of SKI on the TGF-beta mRNA of LLC-PK1 renal tubular epithelial cells, with control group treated with the injection of Rheum officinale extraction. RESULTS: SKI showed the inhibition of TGF-beta mRNA expression of renal tubular epithelial cells, which is dose-dependent and was more obvious than that of the Rheum extractive injection with equal dose of drug. CONCLUSION: The mechanism of delaying chronic renal failure with SKI is achieved by down regulating TGF-beta mRNA expression of renal tubular cells. PMID- 11938870 TI - [Current status and prospect of the treatment of systemic lupus erythematosus]. PMID- 11938871 TI - [Myoglobin and its significance in forensic medicine]. PMID- 11938872 TI - [The development and main achievements of forensic entomology]. AB - The history and main achievements of forensic entomology are introduced. The future of forensic entomology is discussed. PMID- 11938873 TI - [Scanning electronic microscopic observation of vascular corrosion casts of closed diffuse brain injuries in rats]. AB - The brain vascular corrosion casts of closed diffuse brain injuries were studied by SEM in rats. The contraction and spasm of perforating arteriole and sphincter muscle of capillaries in brain stems were observed. The capillaries are about 2-5 microns in diameter, while the normal 10-15 microns. The mechanism of the contraction and its effects were discussed. PMID- 11938874 TI - [Pathological changes of viscera after brain death: an experimental study]. AB - In order to study the viscera changes after brain death while the function of the heart and lung was sustained artificially, 34 cats were used and divided into three groups. The results showed that pathological changes of viscera (heart, lung, liver and kidney) developed, including 1. ischemic cardiomyopathy, endocardiac hemorrhage, focal necrosis, mitochondria and microfilament destruction. 2. pulmonary congestion, edema and parenchymal inflammation. 3. ischemic lesions in the central band of hepatic lobular and ischemic lesions in renal proximal convoluted tubule. PMID- 11938875 TI - [Cardiac pathological study of sudden death due to myocarditis: analysis of 17 cases]. AB - 17 cases of sudden death due to myocarditis were presented. The problem of the classification and mechanism of sudden death is discussed. It is concluded that heart function failure is the main reason for the sudden death of myocarditis. PMID- 11938876 TI - [An immunohistochemical study of basic fibroblast growth factor in early acute myocardial ischemia]. AB - With the use of immunohistochemistry, the expression of basic fibroblast growth factor (bFGF) in early acute myocardial ischemia of rats was studied. It was observed that there was an initial (30 min) rise in bFGF expression in limited ischemic myocardium areas and perimyocardial ischemia areas. The grade of expression of bFGF increased with the continuation of ischemia. The results of image analysis and statistical process suggested that the expression of bFGF reach a high level in ischemic myocardial areas after three hour's of ischemia, significantly higher than in control group and in group B1, but no significant difference in pericardial ischemia areas and normal areas of the same group. In summary, early acute myocardial ischemia is accompanied by rapid and prolonged increase in expression of bFGF with characteristic spatial and temporal kinetic. This finding suggests that the immunohistochemical staining of bFGF provide objective morphologic evidence for the diagnosis of early myocardial ischemia. PMID- 11938877 TI - [A morphological study of human hair tips by scanning electronic microscope (SEM)]. AB - Morphological characteristics of human hair tip was studied by SEM. It was concluded that human hair tips could be classified into eight types. The tip formation, distribution and changes after being trimmed were explored. This study can be used in the mechanism research of human hair growth and the forensic hair examination. PMID- 11938878 TI - [The history of heroin abuse by assaying 6-monoacetylmorphine and morphine in human hair]. AB - The purpose of this study is to conclude the history of heroin abuse by assaying 6-monoacetylmorphine (MAM) and morphine (MOR) in human hair. The hair of heroin abuse was labeled and segmented, then washed and cut into fragments. After hydrolyses and extraction, 6-MAM and MOR in human hair were determined by GC/MS SIM with selected ion monitoring. Results of the segmented hair were analyzed. It provided useful information about the history of heroin abuse (hair growth rate 1 1.5 cm/mon). PMID- 11938879 TI - [The application of solid phase extraction for systematic toxicological analysis of abuse drugs]. AB - A new column was developed in this research. It contains a proprietary bonded silica sorbent that exhibits mixed extraction mechanism. A single-column solid phase extraction procedure was also developed for the screening of acidic, neutral and basic abuse drugs. The recovery of all 7 tested drugs exceeded 60%. The extraction mechanism for different abuse drugs on the new column was explored and was compared with on other columns. It is suggested that this column be effective in systematic toxicological analysis and better than other columns. PMID- 11938880 TI - [Traumatic retina detachment and forensic medical identification]. AB - It is reported the result of the examination and identification of 6 cases with retina detachment. The reasons, characteristics of the traumatic retina detachment and the problems about forensic medical identification were discussed. PMID- 11938881 TI - [Extraction and amplification of DNA from formalin-fixed human tissues]. AB - A method was developed for the extraction of DNA from formalin-fixed human tissues prepared for routine histopathological examination. The extracted DNA was not intact. By increasing the formalin fixation time the necessary amount of intact DNA decreased. The procedure reported here made the final product suitable for quantitative and qualitative analysis and for DNA-typing by PCR technique. PMID- 11938882 TI - [Abuse of "pheticol" and its interference to the analysis of methamphetamine]. AB - The study was to eliminate interference from ephedrine in the analysis of methamphetamine. The extraction procedure for methamphetamine was modified to include an oxidation step (2 ml urine specimen was treated with 0.5 ml of 1 mol/L phosphate buffer (pH 6.8) and 0.5 ml 0.3 mol/L sodium periodate). Results showed that ephedrine could be oxidized in the presence of periodate ions into smaller fragments while leaving methamphetamine intact. It is recommended that specimens be treated with sodium periodate prior to extraction in order to eliminate any interference caused by ephedrine. PMID- 11938883 TI - [The diagnostic significance of "contraction bands" of striated muscle in forensic pathology]. AB - The contraction bands of cardiac and skeletal muscle probes were studied. They were taken from autopsy cases with different death causes. The morphologic characteristics of the contraction bands that were closely related to diagnosis were carefully observed. It was found that the morphological patterns of the contraction bands were various and some were regularly seen for the death with certain causes. The forensic significance of the contraction bands was then discussed and it was suggested that the morphology of contraction bands of the striated muscle correlate to the type and structure of tissues as well as to the property of injuries. PMID- 11938884 TI - [Immunohistochemical research on brain stem injury: expression of heat shock protein 70 in rat brain after traumatic brain stem injury]. AB - The aim of this study was to determine the expression of heat shock protein 70 (HSP70) as molecular chaperones, following traumatic brain stem injury(TBSI). The expression of HSP70 was detected in respective brain regions of rats 1 h, 3 h, 6 h, 12 h, 24 h after brain stem stabbing wound. Numerous HSP70 immunoreactivity positive blood vessel endothelioid and glia cells were found in cerebral, cerebellar cortex and hippocampus after 1 h, with H.E stain unchanged. In the area near the focal of brain stem injury, HSP70 immunoreactivity positive neurons increased significantly after 1 h of injury, reaching the highest level after 3 h, much higher than in other areas. HSP70 immunoreactivity positive cells still existed after 24 h of injury. It was suggested that TBSI cause the defectiveness of protein structure in brain, and induce HSP70 proteolysis of denature protein including misfolding or aggregate. TBSI can be diagnosed regionally by the increasing of HSP70 immunoreactivity positive neurons in the focal of brain stems. PMID- 11938885 TI - [PCR-based detection of HLA-DQ alpha and polymarker loci and their application in forensic science]. AB - Studies were conducted to evaluate the forensic applicability of multiplex amplification of HLA-DQ alpha and PM loci. These loci could be simultaneously typed by reverse dot blot approach where allele specific oligonucleotide probes were immobilized on a nylon membrane strip. Allele and genotype frequencies for HLA-DQ alpha and PM loci were determined in Chinese Han population. The frequency data can be used in individual identification and paternity test. PMID- 11938886 TI - [A program of HLA system for parentage testing and its application]. AB - A software of the HLA blood group system for parentage testing was developed with FoxPro 2.5 system. Chinese prompts helped to follow the process during program running. It is easy to use. 63 cases of parentage testing in our department were calculated, and the results were identical with those of manual calculation. It was believed that this program is useful in the parentage testing. PMID- 11938887 TI - [Immunohistochemical study on ANP in the atria of electrocution]. AB - By the immunohistochemical staining for ANP with LSAB method and the analysis of areas and grays of ANP with computer image analysis, it was found that the ANP granulus in right atria of electrocution were obviously depleted, compared with that in the control groups. The results showed that depletion of ANP granules in right atria could be used as a reference for the forensic diagnosis of electrocution and might benefit for the clinical therapy to the patients suffering from electric injuries. PMID- 11938888 TI - [Discrimination of black inks by FT-Raman spectrometry]. AB - Six brands of black inks on paper were examined by FT-Raman spectrometry. The results showed that the black inks could be discriminated by FT-Raman spectrometry. As the method is easy, quick and undestructive to samples, it is useful in black ink examination. PMID- 11938889 TI - [A survey of the social and psychological factors of the heroin dependence]. AB - The survey studied 100 dependence about their age, marriage, occupation, the way of heroin taking, history of heroin taking, and the reason of the first heroin taking. The data showed that most dependence were young or middle aged men, or people unable to graduate from junior middle school, self-employed laborers and unemployed. Dependence of cadres and people with better education were increasing. The heroin injection is becoming popular. PMID- 11938890 TI - [Analysis of 233 cases of brain injuries in traffic accidents]. AB - 233 cases of brain injuries in traffic accidents were studied. Sex, age and degree of injuries of victims were analyzed. The paper also discussed the relation between the sequel of brain injuries and the level of disability, the time of evaluation, and the determination of nervous obstruction after brain injuries. PMID- 11938891 TI - [Capillary electrophoresis and its uses in the analysis of poisonous substances]. PMID- 11938892 TI - [Progress in the study of astrocyte and its marker glial fibrillary acidic protein in time inference of brain injuries]. PMID- 11938893 TI - [Traumatic cerebral lacunar infarction]. PMID- 11938894 TI - [PCR-based analysis of the D1S80 locus and its application in paternity test]. AB - Allelic data for the D1S80 locus was obtained by using the PCR and subsequent analysis with high-resolution horizontal PAGE technique. In a sample of 184 unrelated Chinese, the D1S80 locus had 75 phenotypes and 20 allelic. The distribution of genotype was in agreement with expected values according to the Hardy-Weinberg equilibrium. Analysis of the D1S80 locus by the PCR has been successfully applied in 150 cases of questioned paternity. PMID- 11938895 TI - [DNA extraction and sex determination of human skeletal remains]. AB - DNA was extracted from human bones remained for 3 to 15 years by using the method developed in our laboratory. A pair of X-Y homologous primer were used to amplify the specific DNA fragments of Amelogenin gene. All the 35 samples yielded correct results. Our method is rapid, sensitive and reliable for the sex determination of skeletal remains. PMID- 11938896 TI - [Distribution of F13A01, FESFPS and vWA loci in Jingpo ethnic group in Yunnan provice]. AB - F13A01 FESFPS and vWA were extracted and amplified using denaturing polyacrylamide gel electrophoresis and silver stain. The distribution of allete frequencies of F13A01, FESFPS and vWA loci in Jingpo ethnic group living in southwestern Yunnan province were investigated. It was observed that there were 8 alleles and 13 genotypes in F13A01 locus; 7 alleles and 18 genotypes in FESFPPS locus; 7 alleles and 21 genotypes in vWA lous. PMID- 11938897 TI - [Determination of chloroquine in human urine]. AB - A GC method was established for qualitative and quantitative determination of chloroquine in human urine. The chloroquine is extracted from 2 ml urine with 2 x 2 ml cyclohexame: chloroform (8:2). The retention time of chlorquine is 9.44 min. The linear of quantitation is 0-50 micrograms/ml. The recovery of the procedure in 25 micrograms g/ml is 87.0%, CV = 7. 9% (n = 5). The limit of the determination is 200 ng/ml. The method can also be used in blood. A case is reported here. In that, we detected chloroquine in urine and blood. The quantitation of chloroquine in urine and blood is 0.475 mg/ml and 3.68 micrograms/ml, respectively. A metabolite of chloroquine-desethylchloroquine is also found in the urine. PMID- 11938898 TI - [Determination of triazolam in human urine by GC/ECD]. AB - A GC/ECD method was established to determine triazolam in human urine qualitatively and quantitatively. The triazolam was extracted from 2 ml urine with 2 x 2 ml chloroform. The retention time of triazolam was 10.74 min. The limit of the determination was 0.5 ng/ml. The recovery of the procedure in 5 ng/ml was 95.98%, CV = 7.85% (n = 5). The limit of quantitation is 2-50 ng/ml. PMID- 11938899 TI - [The polymorphic study of HLA-DRB1 gene in Chinese Liaoning Han with PCR-sequence specific primers]. AB - 8 allele and group specificity at HLA-DRB1 locus were genotyped in 159 unrelated individuals of Liaoning Han, with polymerase chain reaction-sequence specific primers. The range of gene frequencies was from 0.02201-0.23899. Heterozagosity was 83%. 33 out of 36 theoretical genotypes were identified and the result conformed to Hardy-Weinberg equilibrium law. The present method can be used not only in paternity test, individual identification, but also in clinic graft matching between donor and recipient as well as the study of HLA. PMID- 11938900 TI - [Ultrastructure changes of liver autolysis]. AB - Ultra-structure changes of autolysis of hepatocyte were studied using transmission electron microscope(TEM). It was found that autolysis started 0.5 hour after death, with the heterochromatin of nuclei shrinking abnormally and smooth endoplasmic reticulum expanding. The changes developed to distinction after 1 h, and progressed continuously after 2 h, 3 h, 4 h and 5 h. It was found that (1) the mitochondria were swelling, its matrix density decreasing and flocculent dense bodies (FDB) emerging; (2) the rough endoplasmic reticulum and perinuclear cistern were expanding. It was suggested that ultra-structure changes of the autolysis be correlated to the postmortem time. PMID- 11938901 TI - [Forensic clinical identification in medical disputes]. AB - 46 cases of medical disputes were analyzed. The causes of medical disputes were analyzed and the features of forensic identification and medical compensation were discussed. Results showed that 37 cases had causality (cause and effect relation) between medical treatment and negative results. It is concluded that forensic medical identification can play an important role in the civil compensation of medical disputes. PMID- 11938903 TI - [An autopsy analysis on 5 cases of poisoning death with tetramethylenedisulfotertramine]. AB - Pathological changes due to intoxication of tetramethylenedisulphotetramine (TETS) were studied. The results showed that the signs of asphyxia were obvious, so were the congestion and edema of the brain. The spotty and focal hemorrhages in brain stems, multiple myolysis of papillary muscles and contractionbands necrosis of myocardium were observed occasionally. The main clinical symptom of intoxication was frequent clinic--tonic convulsions, which were similar to the grandma epilepsy. The poisoned died mainly in 3 hours. The general poisoning symptoms, LD50, toxicological mechanism and medicolegal expertise were also reviewed. PMID- 11938902 TI - [Pathological studies of 23 cases of thrombus formation in cardiac cavity]. AB - 23 cases of thrombus formation in cardiac cavity were studied. 9 were cased by extensive trauma in soft tissue, 13 by various cardiac diseases. There was no significant difference between trauma and heart disease (p > 0.05). The white thrombus formation was mainly caused by trauma and was often found in the right cardiac cavity (P < 0.001). The mixed thrombus was mainly caused by heart disease and no difference was found between in fight and left cardiac cavity (p > 0.05). The types, positions and causes of thrombus were significantly related (p < 0.001). The causes, pathologic mechanism and forensic identification of thrombus formation in cardiac cavity were discussed. PMID- 11938904 TI - [Analysis of 226 cases of psychosis involved in lawsuit]. AB - 226 cases of psychosis involved in lawsuit were investigated. The problem of forensic identification was discussed. Some suggestions were given to raise the quality of the forensic identification in psychosis. PMID- 11938906 TI - [Studies on voice identification]. AB - The spectrographic method of voice identification was investigated and its application in forensic sciences was discussed. PMID- 11938907 TI - [Current status of the establishment of animal models of mechanical closed craniocerebral injuries]. PMID- 11938908 TI - [Traumatic subarachnoid hemorrhage]. PMID- 11938909 TI - [The property of protein A affinity membrane cartridge and purification of IgG from human plasma]. AB - A new protein A affinity membrane based on composite matrix formed by covalent cross-linking of the glycidyl methylacrylate to softwood cellulose fiber was developed. The study focused on the physical properties and adsorption capacity of the affinity membrane cartridge. The results showed that the relationship between flow-rate and backpressure was linear. When flow-rate was 3 mL/min, backpressure attained 160 kPa. The effects of IgG mass concentration and sampling flow-rate on the binding capacity by protein A cartridge were studied for the optimum purification of IgG. A 21.7 mg/g dry matrix of maximum dynamic binding capacity for IgG was obtained by dynamic adsorption. The cartridge was loaded with human plasma to test its ability to fractionate IgG. The IgG purified from human plasma was analysed by capillary zone electrophoresis (CZE). The results showed that the protein A affinity membrane cartridge had high selectivity for IgG from human plasma. The purity of IgG from human plasma purified by the immobilized protein A membrane cartridge was over 98%. The fast-flow property of the membrane cartridge made from such a matrix and its simplicity in operation provide effective means for purifying immunoglobulins on a relatively large scale. PMID- 11938910 TI - [A study on the binding of cyclodextrins with terbutaline enantiomers by chiral capilary zone electrophoresis]. AB - The recognition mechanism of enantioselectivity in cyclodextrin-modified capillary zone electrophoresis (CD-CZE) on a racemic basic drug terbutaline was discussed. The equilibrium constants of host-guest complexation of terbutaline enantiomers with alpha-cyclodextrin, beta-cyclodextrin, 2,6-dimethyl-beta cycoldextrin, 2,3,6-trimethyl-beta-cyclodextrin and 2-hydroxy-propyl-beta cyclodextrin, and the thermodynamic parameters for chiral recognition procedure were determined respectively. It was found that the matching properties between cavity of cyclodextrins and enantiomers were interrelated with the equilibrium constants of binding complexes and selectivities(a) calculated from the ratio of binding constants of two enantiomers were in the same order of maximum electrophoretic mobility difference between two enantiomers responding with cyclodextrin additive. The experimental value of optimal concentration of cyclodextrin agreed with that calculated from the equation Copt = 1/(K1K2)1/2. From the thermodynamic parameter determination it was shown that the hydrogen bonding interactions between terbutaline enantiomers and 2-hydroxy-propyl-beta cyclodextrin and beta-cyclodextrin should be positive factors. PMID- 11938911 TI - [Optimization of non-gel sieving capillary electrophoretic separation of DNA fragments of hundreds of base pairs]. AB - Non-gel sieving capillary electrophoresis has been employed in the biological sciences for the size-based separation of macromolecules such as nucleic acids. In this paper, four factors i.e. electric field strength, capillary length, capillary diameter and hydroxy-propylmethylcellulose (HPMC) concentration were integratively evaluated to select the optimal condition of separating DNA fragments of a few hundreds of base pairs through orthogonal analysis. Conclusion was made through comprehensive analysis: better separation could be achieved in longer capillary, smaller inside diameter of capillary and less field strength. In practical application, effective separation in short time is important. We preferred to employ 8 g/L HPMC in coated capillary (37 cm x 75 microns i.d.) and electric field strength of 324 V/cm in separating DNA fragments of hundreds of base pairs. From sampling to getting results only a few more than ten minutes were needed in capillary electrophoresis. It needs less amount of sample (a few nanoliters) and shows higher sensitivity (0.1 pmol in UV detection) than polyacrylamide gel electrophoresis (PAGE). pBR322/Msp I fragments were completely resolved in both capillary electrophoresis and PAGE, and four-bases resolution was obtained. When the concentration of the PCR product is too low (< 20 mg/L), PCR reaction system is used as a negative control to make sure of the peak generated by the unpurified PCR products, avoiding interference made by PCR buffer and polymerase. PMID- 11938913 TI - [Advances in chromatography and related techniques in China--status report in 1996-1997]. AB - An overview of the use of chromatography and related techniques in China is given in 1996-1997. This review includes five parts, the statistical results of the scientific papers published in 15 domestic and a few foreign periodicals which are closely related to chromatography and presented on 1997 Beijing Conference and Exhibition on Instrumental Analysis and the 11th National Symposium of Chromatography, about chromatographic theory, GC, LC and CE. One hundred thirty seven references are cited. PMID- 11938912 TI - [Influence of mannitol additive on DNA separation by capillary non-gel sieving electrophoresis]. AB - Polymer solutions with high concentration were employed as sieving buffer in order to obtain satisfactory separation of smaller DNA fragments with length of less than 1,000 base pairs. The concentrated polymer solution had very high viscosity, which was difficult to be pumped into or out of the capillary. Mannitol additive can enhance the sieving ability of hydroxy propylmethylcellulose (HPMC) solution in a Tris-Borate-EDTA background electrolyte. With the existence of mannitol, good separation of PGEM-3Zf(+)/Hae III was able to be obtained in a less concentrated HPMC solution with relatively low viscosity. The optimum mass concentration of mannitol was investigated and it was found that 60 g/L mannitol could give the best results. Mannitol chain could be formed through hydrogen bond among mannitol, HPMC and borate. This kind of mannitol chain shaped the network and decreased the pore size, so the separation was greatly enhanced. PMID- 11938914 TI - [Supercritical fluid extraction and gas chromatographic analysis of four organochlorine pesticides in vegetable]. AB - Supercritical fluid extraction(SFE) of pesticide residues of chlorothalonil, aldrin, dieldrin, and endrin from chives was evaluated. A mixture of vegetable sample: anhydrous magnesium sulfate(2:3) was used for extraction. The extract was analyzed by GC/ECD. The optimum SFE conditions for the pesticides in spiked samples were studied by orthogonal experiment. The chosen conditions were 30.40 MPa, 40 degrees C, 1 min static time, dynamic extraction with 15 mL of CO2, and collection in 3 mL of ethyl acetate. All the pesticides were extracted satisfactorily from their matrix. PMID- 11938915 TI - [Determination of the active flavonoids in silymarine]. AB - The active flavonoids in silymarine extract prominently consist of silychristin, silydianin, silybin (A and B) and isosilybin (A and B). Among these active flavonoids, silybin is the most important one. It is often used to treat liver disorders. An HPLC method for determining thg active flavonoids is described in this paper. The components in the extract can be separated by using a reversed phase system with a C18 column, eluted with methanol and phosphate buffer under gradient conditions, and detected at 280 nm. In comparing with the method of derivatization with 2,4-dinitrophenylhydrazine and detection with UV spectrometry which can only determine the total active flavonoids. The HPLC method not only can separate taxifolin from the flavolignans but also permit individual estimations of silychristin, silydianin, silybin and isosilybin. It has better repeatability. The relative standard deviation is below 2%(n = 5). It can be used for quality control of the crude extracts and the pharmaceutical preparations. PMID- 11938916 TI - [Determination of azathiopurine and 6-mercaptopurine in serum by reversed-phase high performance liquid chromatography]. AB - A method for simultaneous determination of azathiopurine(AZP) and its metabolite 6-mercaptopurine (6-MP) concentration in serum has been developed. Reversed-phase high performance liquid chromatography with gradient elution and dual wavelength detection was employed. After protein precipitation with acetonitrile, the serum sample was evaporated to dryness with a gentle N2 stream at 37 degrees C and 100 microL of the mobile phase were added to dissolve the residue. After centrifugation, 20 microL of the supernatant was injected directly into the HPLC. The detection wavelength was set at 278 nm (AZP) and 325 nm (6-MP). The internal standard was metronidazole. The minimum detectable mass concentration for both AZP and 6-MP in serum was 5 mg/L. The average recoveries were (100.6 +/- 4.2)% for AZP and (102.4 +/- 4.5)% for 6-MP. PMID- 11938917 TI - [Determination of chemical constituents of the volatile oil from Curcuma longa by gas chromatography-mass spectrometry]. AB - The study of chemical constituents of the volatile oil from Curcuma longa is reported. The volatile oil was extracted by steam distillation. Fifteen components in the oil were separated and identified by gas chromatography-mass spectrometry(GC/MS). The results were elucidated based on the NBS standard mass spectral data. The total ion current chromatogram showed their mass fraction by normalization method. alpha-curcumene, alpha-zingiberene, 1,8-cineole and zerumbone, which had been reported before were found in Curcuma longa volatile oil, but 1-(3-cyclopentylpropyl)-2,4-dimethylbenzene, beta-sesquiphellandrene, germacrene etc identified simultaneously in the oil had never been reported. The major chemical constituent of the volatile oil from Curcuma longa is alpha curcumene. PMID- 11938918 TI - [Analysis of phenolic compounds in mineral water by high performance liquid chromatography (HPLC)]. AB - A high performance liquid chromatographic method for the simultaneous analysis of phenolic compounds, such as, phenol, m-cresol, bisphenol A, bisphenol F, 4-tert butylphenol, bisphenol F diglycidyl ether (BFDGE) and bisphenol A diglycidyl ether (BADGE) in mineral water with fluorometric detector is described in this paper. After pretreatment through solid-liquid extraction, the sample was analyzed by HPLC. The operating conditions were Turner C18, 5 microns, column, 150 mm x 4.6 mm i.d., mobile phase with V(ACN): V(water) = 40:60 for 10 min, then linear gradient elution for 25 min up to 80:20 with a flow rate of 1.3 mL/min. Excitation wavelength was set at 275 nm and emmission at 300 nm. The recoveries were 81%-105% and the RSDs were 1.12%-13.21%. Detection limit reached 0.1-0.2 microgram/L. PMID- 11938919 TI - [Determination of resorcinol and salicylic acid in piyanning tincture by high performance liquid chromatography]. AB - A method for the simultaneous determination of resorcinol and salicylic acid in Piyanning tincture by HPLC has been proposed. Operating conditions were Hyppersil ODS column, 4.6 mm x 200 mm, V (methanol): V(water): V(acetic acid) = 50:50:0.9 mobile phase and UV detection at 285 nm. The linear ranges of the method were 0.05-0.25 g/L(r = 1.000) for resorcinol and 0.025-0.127 g/L(r = 1.000) for salicylic acid. The limits of detection were both 0.2 mg/L at a signal-to-noise of 3. The assay method was capable to resolve resorcinol and salicylic acid from their impurities. PMID- 11938920 TI - [Determination of lactitol in mice serum by high performance liquid chromatography]. AB - An HPLC method for the determination of lactitol in mice serum was developed. A liquid chromatograph equipped with a refractive index detector and a Waters Sugar Pakl column was used. The column temperature was maintained at 90 degrees C and the mobile phase was redistilled water. One milliliter of the serum sample was treated with 3.0 mL methanol. The precipitate was centrifuged for 15 min at 10,000 r/min. Two milliliters of the supernatant was dried on a water bath at 100 degrees C. The residue was dissolved with 1.0 mL water and 10 microL of which was injected. The average recovery was 98.6%. The detection limit was 0.1 microgram (S/N = 3.75). The linear range was 0-600 mg/L (r = 0.9992). Both the RSD's within day and between days were below 1.0% (n = 5). PMID- 11938921 TI - [Determination of trace 4,4'-diaminobiphenyl, 4-nitrophenol and phenol in environmental water by high performance liquid chromatography]. AB - A method for simultaneous determination of trace 4,4'-diaminobiphenyl, 4 nitrophenol and phenol in waste water by high performance liquid chromatography with spectrophotometric detection is reported. These components were separated on a Phenomenex Spherex C18 column with V(acetonitrile): V(ether): V(50 mmol.L-1 acetate) = 12:10:78 buffer (pH 6.0) as mobile phase and UV detection at 275 nm. The detection limits were 0.14, 0.19 and 0.08 ng for 4,4'-diaminobiphenyl, 4 nitrophenol and phenol, respectively, when the ratio of signal to noise was 2. The relative standard deviation was < 2.2 (n = 6). This method is sensitive and has been applied to the analysis of environmental water samples with satisfactory results. PMID- 11938922 TI - [Simultaneous determination of aspartame and amino acids in fermented milk beverages by HPLC]. AB - An RP-HPLC method for the simultaneous determination of aspartame and amino acids in fermented milk beverages were established. Samples were prepared by mixing with methanol at a ratio of 1:1 and centrifuged at 4,000 r/min for 15 min. Ten microL supernatant was moved into a sample tube, dried and derivatized according to PICO-TAG procedure developed by Waters. A Novapak C18 column (3.9 mm x 150 mm) was used instead of PICO TAG column and the gradient elution program was modified correspondingly. Column temperature was maintained at 38 degrees C and the components were detected at 254 nm. Linearity for aspartame at the range in 1-100 mg/L is A = 333 C + 20 with r = 0.9996 where A is the integrated area of chromatographic peak of aspartame and C is the corresponding mass concentration. Repeatability for 8 injections was tested and the RSD of 3.2% was obtained. The recovery of aspartame for 5 samples with the method ranged from 94.2% to 98.7%. PMID- 11938924 TI - [Determination of Cd, Cu, Pb, Hg by reversed-phase high performance liquid chromatography]. AB - The chromatographic behaviors of Me(n+)-Dz (dithizone) have been studied with RP HPLC. A method for the determination of Cd, Cu, Pb and Hg has been established. The chromatographic conditions were: column: Shim-pack CLC-ODS, 150 mm x 6.0 mm i.d.; mobile phase: V(methanol): V(water): V (chloroform) (containing volume fraction 1% triethylamine) = 80:12:8; flow rate: 1 mL/min; column temperature: 35 degrees C; detection wavelength: 254 nm. The linear ranges were from 0.01 mg/mL to 2.0 mg/mL with correlation coefficients of 0.9993-0.9998. The detection limits of Cd, Cu, Pb, Hg were from 2.4 micrograms/L to 5.0 micrograms/L. The RSDs were in the range from 1.8% to 9.7%, and the recoveries ranged from 94% to 103% (except Hg). The method has been applied to the analysis of hair. PMID- 11938923 TI - [Study on the analytical method of s-triazine herbicide residues in soil and maize]. AB - The sample was extracted with a mixture of V(acetonitrile): V(methanol) = 1:1. After filtration, the extract was defatted with petroleum ether and then it was passed through a short C18 column for pretreatment and analysed by Nova-Pak C18 column, 150 mm x 3.9 mm i.d., with V(MeOH): V(H2O) = 55:45 as mobile phase. The recoveries were 82.4%-99.8% for cyanazine, 85.6%-102.3% for atrazine and 89.1% 108.4% for simetryne. PMID- 11938925 TI - [Quantitative determination of fenfluramine hydrochloride and its tablets by high performance liquid chromatography]. AB - Fenfluramine hydrochloride and its tablets were determined with high performance liquid chromatography by using an analytical column (25 cm x 0.46 cm i.d.) packed with Hypersil BDS C18 (5 microns), a UV detector at 264 nm and V(acetonitrile): V(0.4 mol/L ammonium acetate): V(triethylamine) = 30:70:2 as eluent. The quantitative determination was performed with caffeine as the internal standard. The internal standard method showed good linearity in the mass concentration range of 0.1-0.5 g/L with r = 0.9999. The average recovery of fenfluramine tablets was 99.98%. This method is simple, rapid, sensitive and reproducible. PMID- 11938926 TI - [Studies on the antifertility effect of bis-trichloromethyl sulfone and its reversibility in male rats]. AB - Bis-trichloromethyl sulfone(BTS) given p.o. to male Sprague-Dawley rats at a dose of 10 mg.kg-1.d-1, 6 days a week for 8 weeks(or 30 mg.kg-1.d-1 for 4 weeks), was found to lead to infertility. The density and, in particular, the motility of the spermatozoa in the cauda epididymidis were significantly lower than those of the controls (P < 0.001). The body weight, the sex behavior, the blood testosterone level and the histology of important organs were not significantly different from those of the control rats. Fertility started to recover 4 weeks after cessation of medication and was entirely recovered 2 more weeks later in rats having received BTS for 8 weeks at the dose of 30 mg.kg-1.d-1. In vitro study showed that BTS at a concentration of 0.31 mg.ml-1 would make rat spermatozoa immobile within 20 seconds. These results indicate that BTS has both antifertility effect in male rats and direct inhibitory effect on rat spermatozoa. We conclude that nontoxic doses of BTS can induce reversible infertility in male rats. PMID- 11938927 TI - [Effects of methotrexate on early pregnancy in mice]. AB - Single s.c. injection of MTX at dosages of 8, 24 and 72 mg.kg-1 on day 7 of gestation was found to have a feticide effect in mice. The mortality of fetuses on day 13 of pregnancy was 18.0%, 55.5% and 60.3%, respectively. Treatment with MTX caused a decrease of serum progesterone level on day 13 of pregnancy. Treatment with MTX at a dosage of 24 mg.kg-1 on day 7-9 of gestation terminated total early pregnancy. MTX combined with hCG 10 U/mouse or progesterone 0.2 mg/mouse was shown to have antagonistic effect to terminate early pregnancy. Synergism of RU486 combined with MTX on termination of early pregnancy in mice was observed. PMID- 11938928 TI - [Protective effect of hyperin against cerebral ischemia-reperfusion injury]. AB - The protective effect of hyperin (Hyp) against cerebral ischemia-reperfusion injury was studied. On the cerebral ischemia-reperfusion model in mice, Hyp (50, 100 mg.kg-1, i.p.) was shown to markedly and dose-dependently inhibit the decrease of lactate dehydrogenase (LDH) in cerebrum and improve the learning and memory impairment on the step down test. On the four-vessel occlusion model in rats, Hyp(50 and 100 mg.kg-1, i.p.) significantly reduced the decreases of glutathione peroxidase(GSH-Px), superoxide dismutase(SOD) and LDH activities in the cerebrum. Hyp was also shown to inhibit the increase of nitric oxide (NO) and malondialdehyde (MDA) contents in the cerebrum and promote the recovery of EEG activities. These results suggest that Hyp has protective effect against cerebral ischemia-reperfusion injury via attenuating free radical and NO. PMID- 11938930 TI - [Synthesis of baicalin derivatives and evaluation of their anti-human immunodeficiency virus (HIV-1) activity]. AB - The aglycone baicalien(1) and the key intermediate 5,6-O-dibenzyl-baicalien(3) were prepared from baicalin in order to evaluate the influence of different glycosyloxies linked to baicalien on anti-HIV activity. Four new flavone glycosides namely 5-hydroxyl-6-O-benzyl-flavone-7-beta-D-glucoside(12), 5 hydroxyl-6-O-benzyl-flavone-7-beta-D-galactoside(13), 5-hydroxyl-6-O-benzyl flavone-7-alpha-D-mannoside(14) and 5-hydroxyl-6-O-benzyl-flavone-7-alpha-D arabinoside (15) were synthesized by condensation of the corresponding protected glycosyl bromides with (3). Biological activity test showed that (a) both baicalin and baicalien inhibited HIV-1 RT; (b) the 6-hydroxyl substitution of baicalin and baicalien is necessary for their inhibitory activity on HIV-1 RT; (c) the HIV-1 inhibitory activity and cytotoxicity of baicalien were higher than those of baicalin, the two compounds were found to have almost identical therapeutic index. PMID- 11938929 TI - [Anti-invasion activity of several plant-originated anticancer drugs with different mechanism of action]. AB - The antitumor and anti-invasion activities of taxol, harringtonine, homoharringtonine and camptothecin on highly metastatic melanoma B16-BL6 and human fibrosarcoma HT-1080 cells were reported in this paper. Results demonstrated that taxol, harringtonine, homoharringtonine and camptothecin exhibited significant inhibition of cell growth of B16-BL6 and HT-1080 cells. Taxol, harringtonine and homoharringtonine were also found to be effective for the inhibition of cell invasion and migration of B16-BL6 cells, but camptothecin showed basically no effect at the indicated concentration. PMID- 11938931 TI - [Synthesis and biological activities of 1-alkyl-1-benzyl-4-(3-chloro-2 hydroxy)propyl piperazinium halides]. AB - Seven 1-alkyl-1-benzyl-4-(3-chloro-2-hydroxy) propyl piperazinium halide hydrochloride were synthesized. Preliminary biological activity test showed that the presence of electro-attracting group in the benzyl distinctly increased the analgesic and anti-inflammatory activities of the compounds. For example, the analgesic activity of Ie and the anti-inflammatory activities of Id and Ie were all higher than those of Ia and Ib. The different anion of quaternary ammonium have some effect on the biological activity. However, this effect is not regular. PMID- 11938932 TI - [A limonoid from the Chinese drug dong-feng-jie (Atalantia buxifolia)]. AB - A new limonoid was isolated from the ethanol extract of dried roots and stems of Atalantia buxifolia (Poir) Oliv.. Based on chemical properties and spectral analysis, including 1H- and 13CNMR, DEPT, MS, IR and UV spectra, its structure have been identified as: 6-deoxy-6 alpha-acetoxyatalantin acetate. This compound was isolated from this plant for the first time. PMID- 11938933 TI - [A new saikosaponin from Bupleurum Chinese DC]. AB - Bupleurum chinese DC. is a well-known and very important traditional Chinese drug. It is often used to treat common cold with fever, alternating chill and fever, the feeling of fullness and oppression in the chest. A new saikosaponin was isolated from Bupleurum chinese DC. and its structure was identified as 3 beta, 16 beta, 28-trihydroxy-11-alpha-methoxy-olean-12-ene-3-O-beta-D-glucosyl-(1 ->3)-beta D-fucoside on the basis of chemical and spectral evidence. In addition, six known saikosaponins were isolated and characterized as saikosaponin a, saikosaponin d, saikosaponin c, saikosaponin f, saikosaponin b3 and saikosaponin b2. All these compounds were isolated from Bupleurum chinese DC. for the first time. PMID- 11938934 TI - [RP-HPLC method for determination of isorhynchophylline in plasma and its pharmacokinetics in rabbits]. AB - A reversed phase HPLC method for determination of isorhnchophylline (IRHY) in rabbit plasma was developed. A Shimadzu LC-10A liquid chromatograph equipped with a Shimadzu SPD-10A multiwavelength detector and a Chromatopac LC-10A data processor were used. The analytical column was a 4.6 mm x 150 mm CLC-ODS. The mobile phase was a mixture of methanol-water (95:5), the flow rate was 1 ml.min 1. Detection was performed at UV 254 nm. The calibration curve was linear in the range from 0.016 to 16 micrograms.ml-1 with gamma = 0.9990. The lowest detection limit was 0.016 microgram.ml-1. The precision and accuracy of within-day and day to-day ranged from 3.8% to 6.9%. The recoveries were 80.5% to 85.1%. The drug concentrtion-time curves of IRHY in rabbits after i.v. of 2 mg.kg-1 and 5 mg.kg-1 were shown to fit a two-compartment open model with half-times of 1.32 +/- 0.6 h and 1.25 +/- 0.09 h respectively. The absorption was fast after intraduodenal administration of IRHY (2 mg.kg-1 and 5 mg.kg-1) to rabbits with half-times of 1.75 +/- 0.18 h and 1.26 +/- 0.20 h, respectively. The bioavailability of IRHY was about 42.4% to 69.4%. The results showed that the method was simple and fast. The absorption and elimination were fast atfer intraduodenal administration or i.v. of IRHY to rabbits. PMID- 11938935 TI - [Effect of heparinoid on the proliferation of rabbit vascular smooth muscle cells in vitro]. AB - Proliferation of vascular smooth muscle cells(VSMCs) represents an important event in vascular lesion formation. In the present study, we investigated whether heparinoid abstracted from porcine duodenum inhibits the proliferation of rabbit aortic VSMCs in vitro, using the method of colorimetric MTT (tetrazolium). Our results showed that heparinoid at 1.6-0.05 mg.ml-1 significantly inhibits VSMCs proliferation induced by fetal calf serum(FCS, 10%), basic fibroblast growth factor(bFGF, 50 ng.ml-1) or interleukin-1(IL-1, 50 u.ml-1). In inducing with bFGF, the inhibitive effect of heparinoid was more potent than that of heparin, while in inducing with FCS or IL-1, the inhibitive effect of heparin was more potent than that of heparinoid. Preincubation with heparinoid at 0.2-0.05 mg.ml 1, the vascular smooth muscle cell proliferation induced by FCS was also inhibited. Our results imply that heparinoid may be useful to protect the atherosclerosis and angioplasty restenosis. PMID- 11938936 TI - [Separation of human bladder cancer cells from bone marrow with a kind of immunomagnetic microspheres]. AB - By binding a monoclonal antibody BDI-1 to polystyrene magnetic micospheres using physical and chemical methods together, a kind of immunomagnetic microspheres (IMMS), which can specifically bind target cells and give it magnetic response, was sucessfully constituted. The results of indirect immunofluorescence and cell binding suggest that IMMS can effectively bind target cells while do not have non specific binding activity. The influence of antibody added and the ratio between IMMS and cells were also studied. After purging cancer cells from bone marrow of mice with IMMS, the cancer cells were effectively removed while most of the bone marrow cells were recovered. PMID- 11938937 TI - [Study on the rifampicin polylactic acid microspheres for lung targeting]. AB - In this paper, the effects of different variables on the preparation of polylactic acid microspheres (PLA-MS) were studied. The optimized preparation conditions of rifampicin polylactic acid microspheres (RFP-PLA-MS) were aquired through orthogonal test. The paddle method was used to study the drug release properties of RFP-PLA-MS. Stability of RFP-PLA-MS at different temperatures was also studied. Pharmacokinetic and tissue distribution of RFP-PLA-MS after intravenous administration were carried out in rabbits. The experiments revealed that the RFP-PLA-MS was regular in its morphology with a mean diameter of 9.00 +/ 4.08 microns. The drug loading was 16.0% and encapsulation efficiency was 31.9%. The release properties could be expressed by the following equation: Q = 20.77 + 10.12 T 1/2 (gamma = 0.9892). The RFP-PLA-MS was stable after stored at 4 degrees C and room temperature under desiccated condition for three months. RFP-PLA-MS showed a combination of lung targeting and sustained drug release in experiments on rabbits. PMID- 11938938 TI - [Effects of acetylsalvianolic acid A on arachidonic acid metabolism in platelets]. AB - Acetylsalvianolic acid A (ASAA) has been shown to be an antiplatelet compound. Our studies with RIA showed that: at the dosage of inhibiting platelet aggregation, ASAA was found to inhibit the formation of cyclooxygenase pathway metabolites TXB2 in platelets. In addition, ASAA was also shown to promote the formation of 6-keto-PGF1 alpha in rabbit aortic rings in vitro. The above results suggest that ASAA may inhibit the proaggregator TXA2 formation, promote mildly the antiaggregator PGI2 generation and probably, whereby, exert its antiplatelet activity. PMID- 11938939 TI - [Absolute configuration of the diastereoisomer of 2-amino-3-(1,2 dicarboxyethylthio) propanoic acid from Amanita pantherina]. AB - Four stereoisomers of 2-amino-3-(1,2-dicarboxyethylthio) propanoic acid were prepared by reaction of L- and D-cysteine with fumaric acid. The absolute configuration of the diastereoisomer of 2-amino-3-(1,2-dicarboxyethylthio) propanoic acid from Amanita pantherina were assigned as (2R, 1'R) and (2R, 1'S) by analysis of the optical properties. Pharmacological tests showed that all of the four stereoisomers inhibited the depolarization of NMDA on spinal motorneurones in newborn rats, The inhibition intensity of L-A, D-A and D-B were higher than that of L-B. PMID- 11938940 TI - [Studies on synthesis and anti-HIV RT activity of hypericin and ethylhypericin]. AB - Condensed polycyclic anthraquinone hypericin and its analogues showed activity against several retroviruses, including human immunodeficiency virus (HIV). Another condensed polycyclic anthraquinone ethylhypericin was synthesized from butanone in order to compare its biological activity with hypericin. Biological activity showed that ethylhypericin was slightly more effective than hypericin in HIV-RT inhibitory test. PMID- 11938941 TI - [Synthesis and biological activity of 2,3-benzopyrone analogs]. AB - Several 2, 3-benzopyrone analogs were synthesized for evaluating their K(+) channel and anticancer activities. The Friedel-Crafts reaction was taken place with some replacement phenyl acetic acid or its methyl ester and vanillin as reactants in the condition of Ac2O/ZnCl2. All products were confirmed by means of MS and 1H, 13CNMR spectra. Compounds 2 and 3 showed obvious activities on the K(+)-channel and anticancer screening, while the activity of compound 5 was weak. Compound 4 was not yet tested. The primary result indicate that the potential activity of such compounds deserves further study. PMID- 11938942 TI - [Gene regulation of apoptosis and study of anti-neuronal apoptotic agents]. PMID- 11938943 TI - [Effects of some mexiletine derivatives on alpha 1-adrenoceptors]. AB - Using [3H]-WB 4101 binding assay in rat cerebral cortex membranes, effects of 18 mexiletine derivatives on alpha 1-adrenoceptors were studied in order to find new antihypertensive alpha 1 receptor blocking agents. The results showed that 6 of them showed significant affinities to alpha 1-adrenoceptors in rat cerebral cortex membranes. Some structure-activity relationship were found, among them only the compounds with chiral carbon showed high affinity to alpha 1 adrenoceptor. The affinity of compound M-85001 (pKi = 6.51) was shown to be higher than that of tolazoline. In the rat anococcygeal muscle, compound M-85001 competitively antagonized phenylephrine-induced contraction with pA2 value of 6.86 which is similar to its pKi value in the binding assay. These findings may be of significance in the search for novel class of alpha 1 receptor antagonists. PMID- 11938944 TI - [Effects of dopexamine on cardiac hemodynamics during myocardial ischemia in rats and in comparison with fenoldopam and procaterol]. AB - The effect of dopexamine hydrochloride(DPX) on cardiac hemodynamics during acute myocardial ischemia (AMI) was studied in rats and compared with those of fenoldopam hydrochloride(FODA) and procaterol hydrochloride (PCT). The results showed that i.v. DPX remarkably attenuated the increases of total peripheral resistance(TPR) and left ventricular end-diastolic pressure and the decreases of + dp/dt max caused by ischemia, while DPX showed no significant effect on mean arterial pressure(MAP). Although FODA was more potent than DPX in attenuating the increase of TPR and the decrease of myocardial contractility caused by ischemia, it significantly decreased MAP. The effects of PCT on all these variables were the smallest among the 3 agents. The results suggest that DPX has a better prospect in the treatment of ischemic heart disease in view of its actions of decreasing TPR, mild increase of myocardial contractility, without MAP decrease and myocardial oxygen consumption increase. PMID- 11938945 TI - [The synthesis and immunosuppressive effects of steroid-peptide linkers]. AB - Hydrocortisone was coupled with urotoxin tripeptide UTP-A, UTP-B and UTP-C respectively yielding 4 linkers. Their bioactivities such as prolongation of heterotopic transplanted cardiac tissue survival, inhibitory effects on phagocytosis of mouse peritoneal macrophages and the influence on Con A induced proliferation of spleen lymphocytes of mouse were observed. Compared with UTP-A, UTP-B, UTP-C or hydrocortisone the linkers were more potent immunosuppressants. The results suggest that the linker of steroid-peptide may simulate the permissive action. PMID- 11938946 TI - [A new method for study of the effect of drugs on cancer cells--31PNMR with perfused cell system]. AB - 31P NMR was performed for the studies on the metabolic changes of endogenous phosphorus-containing molecules in KB and HCT-8 cells exposed to taxol at a concentration of 10(-6) mol.L-1. Using the perfusion method, the cells can be detected for a longer time by NMR, so as to give continuous spectra of the two cell-lines during the perfusion with and without the drug. The spectra showed that the levels of ATP peaks for both cells enhanced during the perfusion with the drug, but the change of the level is more prominent in KB cells than in HCT-8 cells. This shows that KB cells are more sensitive to taxol than HCT-8 cells. This is in coincident with the result in the cytotoxicity studies. However, the experiment using vincristine at the same concentrations demonstrated that the level of the ATP peak was not changed significantly. PMID- 11938947 TI - [Synthesis and bioaction of 2-alkyl-4(1H)-quinolone]. AB - Five quinolone alkaloids (IVa, Va, c, d, g) including two new compounds (IVa, Vc) from Evodia rutaecarpa and nine analogs are synthesized in good yield by using acid-catalyzed condensation of a series of 3-oxoalkkanoic acid esters with aniline and further methylized with methyl iodide. The other analogue Vh is prepared through the reaction of lithium enolate methyl ketone with N methylisatoic anhydride. Eight compounds (IVa, d, f; Vb, c, d, f, h) were synthesized for the first time. Pharmacological studies showed that these compounds have vasoconstriction inhibiting and antiulcer effects. Compound IVb also has cytotoxic effect. PMID- 11938948 TI - [Studies on diterpenoids from the roots of Euphorbia ebracteolata]. AB - Euphorbia ebractolata has been used as insecticide and tuberculocide. Four crystalline compounds were isolated from the root of this plant. They were characterized by their physico-chemical properties and spectral data as: 3-acety alpha-amyrin (I), Jolkinolide B (II), ebracteolatanolide A (III) and ebracteolatanolide B (IV). I and II were isolated for the first time from E. ebracteolata Hayata. III and IV are new compounds. PMID- 11938949 TI - [Effects of phenylalanine, sucrose and mannitol on the growth and production of taxol, baccatin III and 10-deacetylbaccatin III in suspension cells of Taxus media]. AB - The effects of phenylalanine, sucrose and mannitol on the cell growth and the production of taxol, baccatin III and 10-deacetylbaccatin III in the suspension cells of Taxus media were studied. The results showed that phenylalanine 1.0 mmol.L-1 or 2.0 mmol.L-1 initially added into the medium, and sucrose 73.0 mmol.L 1 and mannitol 173.3 mmol.L-1 added into the medium at the 28th d of culture strongly promoted the cell growth and the formation of the three taxanes in the suspension cells. Compared with those of the control, the cell biomass of the treatments supplemented with phenylalanine and added with sucrose and mannitol at the 28th d of culture increased by 0.6-0.8-fold, taxol yield by 9-10-fold, baccatin III yield by 2.5-3.0-fold, and 10-deacetylbaccatin III yield by 7-fold. Addition of sucrose 73.0 mmol.L-1 at the 28th d of culture significantly promoted the cell growth, but showed little effect on the contents of the three taxanes in the suspension cultures. PMID- 11938950 TI - [A reversed phase HPLC method with pre-column derivatization to determine propafenone enantiomers in human plasma]. AB - A reversed phase HPLC method to stereoselectively determine enantiomers of propafenone in human plasma has been developed. After extraction of 1 ml plasma with 3 ml of n-C6H14:CH2Cl2:2-C3H7OH (100:50:5, v/v) and dried under N2, the enantiomers of propafenone were allowed to react with homochiral S(+)-1-(1 naphthyl) ethyl isocyanate (5.66 micrograms in 60 microliters CH2Cl2) at RT for 30 min, to give the diastereomeric derivatives. Their separation was achieved using HPLC with a C18-column and UV-detection (220 nm) and the enantiomeric ratios were measured. The concentration of each enantiomer were then calculated using the enantiomeric ratios and the racemic propafenone concentrations previously measured. This method after validation procedures has been applied to the multisample analyses of a human pharmacokinetic study with 10 healthy volunteers after an oral dose of 300 mg propafenone hydrochloride. The method was shown to be sensitive (7.5 ng.ml-1), convenient and reproducible. PMID- 11938951 TI - [Determination of 6 isoflavonoids in the hairy root cultures of Astragalus membranaceus by HPLC]. AB - An accurate RP-HPLC method for the determination of 10-hydroxy-3, 9 dimethoxypeterocarpan, (3R) 8,2'-dihydroxy-7,4'-dimethoxyisoflavan, formonenetin (7-hydroxy-4'-methoxyisoflavone), 8, 3'-dihydroxy-7, 4'-dimethoxy-isoflavone, 2' hydroxy-3', 4'-dimethoxyisoflavan-7-O-glucopyranoside and alycosin (7,3' dihydroxy-4'-methoxyisoflavone) in the hairy root cultures of Astragalus membranaceus was developed. Among them, 10-hydroxy-3, 9-dimethoxypeterocarpan, (3R) 8,2'-dihydroxy-7,4'-dimethoxyisoflavan and 8,3'-dihydroxy-7,4' dimethoxyisoflavone were new compounds in Astragalus membranaceus. The analytical column was Nucleosil C18. The mobile phases were methanol-water = 3:2 and 1:1 (v/v). The detection wavelengths were 254 nm and 280 nm. Calibration graphs were rectilinear between 2.5 micrograms and 12.5 micrograms. The recoveries were between 96.47% and 103.33%. The relative standard deviations (RSDs) of measurement precision test were 2.57%-6.52%. The flavonoid contents in the hairy root cultures were between 0.0005% and 0.0065% (dry weight). PMID- 11938952 TI - [In vitro release kinetics and hypoglycemic effect on diabetic rats after oral administration of insulin loaded nanoparticles]. AB - The insulin-loaded polyalkylcyanoacrylate nanoparticles (INS-NP) were made with Dextran 70 as the stabilizer. The mean diameter of INS-NP was 252.4 nm with a poly dispersity of 0.005. The associating ratio of insulin to the nanoparticles reached 70.1% +/- 2.3%, while the loading capacity was 0.14 u.mg-1. Studies on in vitro release kinetics showed that release profiles can be well modelled using a biexponential function. The burst effect was obvious, and a faster release was observed in acidic media. After various doses of INS-NP were intragastrically given to diabetic rats, significant decrease of glucose level was achieved in the 10 and 20 u.kg-1 groups, with no significant difference between these two doses. The relative bioavailability after p.o. administration of INS-NP 10 u.kg-1 over s.c. administration of insulin solution 1 u.kg-1 was 7.58% calculated by the area over the curve of glucose level (%) versus time profiles. The correlation was obvious between the % of insulin released at pH 7.0 and the % of glucose decreased within the first 7 hours. Hence, an oral insulin preparation with rather high bioavailability was provided in this study, and its shorter effective time will make it more convenient for the control of the glucose level in clinics. PMID- 11938953 TI - [Recent development in quinolone compounds antitumor quinolones]. PMID- 11938954 TI - [Protective effects of schisanhenol against oxygen free radical induced injury of rat cerebral mitochondria and synaptosomes]. AB - The cerebral tissue injury induced by ischemia and reperfusion or trauma has been considered to be due to over production of oxygen free radicals (OFRs). The aim of this study was to evaluate the effects of schisanhenol (Sal) on Fe(2+) cysteine (Cys) induced injury of rat cerebral mitochondria and synaptosomes in vitro. Incubation of cerebral mitochondria or synaptosomes with Fe(2+)-Cys at 37 degrees C resulted in an increase of malondialdehyde (MDA) formation and decrease of ATPase activity. Sal(10(-4) mol.L-1) completely inhibited Fe(2+)-Cys induced increase of MDA formation of mitochondria and synaptosomes as well as the loss of ATPase activity of mitochondria. The swelling of mitochondria and reduction of membrane fluidity of mitochondria and synaptosomes induced by Fe(2+)-Cys were also prevented by Sal. Sal(10(-5) mol.L-1) was shown to significantly inhibit the decrease of synaptosomal GSH content induced by H2O2. The electron micrographs also showed that Sal markedly reduced the pathological damage of mitochondria and synaptosomes induced by Fe(2+)-Cys. The results suggest that Sal has protective action against Fe(2+)-Cys induced injury of rat cerebral mitochondria and synaptosomes. PMID- 11938955 TI - [Antagonism of the novel cholinolytic tricyclopinate on nicotinic and muscarinic cholinergic receptors]. AB - Tricyclopinate hydrochloride(TCPN.HCl) and methiodide(TCPN.CH3I) have been identified as new chemical entities. The effects of these two compounds on central and peripheral nicotinic and muscarinic cholinergic receptor activities were investigated. Excitation of the central nicotinic receptors by nicotine produced convulsions in mice. The dose-response curves of nicotine for producing convulsions were shifted rightward by TCPN.CH3I in a paralled manner. Excitation of the central muscarinic receptors by arecoline produced tremors in mice. TCPN.HCl was shown to prevent arecoline-induced tremors. In isolated guinea-pig ileum preparations. TCPN.HCl was found to antagonize nicotine-induced contractions due to excitation of ganglionic nicotinic receptors. In xenopus laevis embryo neuron-muscle co-cultured cells, TCPN.HCl blocked spontaneous miniature endplate currents, and showed preference to blocking the nicotinic receptor ion channels, which had a long open time, and high current amplitude. The anticholinergic effects of TCPN.CH3I were weaker than those of TCPN.HCl. In conclusion, TCPN.HCl has potent effects against nicotinic and muscarinic receptors in the central and periphery nervous systems. PMID- 11938956 TI - [Evaluation of penicilloyl proteins of allergic impurity in gene engineering drugs]. AB - It is well known that penicillins are necessary for prevention of contamination during cell culture or fermentation in the production of gene engineering drugs. As penicillins are easily combined with proteins forming allergic impurities, the benzyl penicilloyl proteins in aqueous solution, an evaluation system should be established for control of the gene engineering drugs. An enzyme linked immunosorbent assay (ELISA) with high specificity and high sensitivity in vitro, by which the conjugated penicillins of 0.3 ppm in a sample could be detected and a passive cutaneous anaphylaxis (PCA) test, a classical method for evaluation of allergic reaction in vivo are integral parts of this system. Some domestic gene engineering drugs such as erythropoietin (EPO), G-CSF, GM-CSF and 125SerIL-2 are evaluated with this system. No matter whether the expression products was produced in secretion manner or in inclusion body, it was possible that some residues of allergic impurities may remain in the finished products if an unsuitable process of isolation and purification was used. The maximum absolute content of the conjugated penicillins in a sample was up to about one per thousand. PMID- 11938957 TI - [Effect of neurotropin on brain edema induced by permanent focal cerebral ischemia in rats and collateral ventricular injection of carrageenan in mice]. AB - Neurotropin, an inhibitor of the kallikrein-kinin system, has been used as an analgesic and anti-allergic drug in the clinic. The aims of this study are to evaluate the effect of neurotropin on brain edema induced by permanent focal cerebral ischemia in rats and collateral ventricular injection of carrageenan in mice and its mechanism. Drugs were given i.v. 15 minutes after middle cerebral artery occlusion in rats and immediately after carrageenan injection in mice. Brain water content was determined by wet/dry weight ratio. Results showed that neurotropin could reduce the brain edema induced by permanent focal cerebral ischemia in rats and collateral ventricular injection of carrageenan in mice at doses of 3.0, 6.0, 30.0 and 30.0 NU.kg-1 body weight. It also suggests that neurotropin reduced ischemic brain edema via inhibiting the kallikrein-kinin system. PMID- 11938958 TI - [Effects of beta-endorphin on glutamate neurotoxicity]. AB - The effects of beta-endorphin(beta-End) on monosodium glutamate(MSG) neurotoxicity were studied via morphological observation and image analysis of neuronal areas, together with the determination of intracellular free calcium concentration ([Ca2+]i) in single neuron and radioimmunoassay for beta-End contents. beta-End(1.0 mg.kg-1, s.c.) was found to obviously aggravate the neuronal injury in the arcuate nucleus of hypothalamus induced by MSG(0.5 g.kg-1, s.c.). Just as MSG increased [Ca2+]i significantly, beta-End(2.0 g.L-1) itself also increased it, though the extent of elevation was smaller than that of MSG(17.0 mg.L-1). The obvious changes of [Ca2+]i induced by both MSG and beta-End were partially reversed after pretreatment with verapamil. On the other hand, the content of beta-End in different areas of the brain were augmented following the addition of MSG and further elevated by morphine treatment. These findings suggest that the mechanisms of the enhancing effect of beta-End on glutamate neurotoxicity were linked to the aggravation on the disruption of intracellular Ca2+ homeostasis induced by MSG. Moreover, the fact that opioids promote beta-End release induced by MSG may be involved in the mechanisms as well. PMID- 11938959 TI - [Effects of daurisoline and its three optical isomers on ischemic injury in cultured pheochromocytoma (PC12) cells]. AB - In this study, the protective effects of (-)-R.R-daurisoline and its three optical isomers on ischemic injury in cultured PC12 cells induced by treating cells with NaCN in glucose-free medium were investigated. Cell viability was measured using MTT assay. The results indicated that these compounds, especially (-)-S.R and (+)-R.S isomers were found substantially to attenuate ischemic injury in PC12 cells in a dose-dependent manner. The IC50 values of (-)-R.R, (-)-S.R, (+)-R.S and (+)-S.S isomers were shown to be 18.6 x 10(-6), 2.4 x 10(-6), 5.9 x 10(-6) and 90 x 10(-6) mol.L-1, respectively. Intracellular free Ca2+ concentration in PC12 cells was measured using AR-CM-MIC cation measurement system with Fura-2/AM as Ca2+ fluorescent indicator. (-)-R.R-daurisoline and its three optical isomers: (-)-S.R, (+)-R.S and (-)-S.S were found to markedly inhibit the increase of cytosolic free Ca2+ concentration induced by NaCN (20 mmol.L-1) in a dose-dependent manner. Their IC50 were found to be 3.55 x 10(-6), 0.59 x 10(-6), 1.29 x 10(-6) and 24.3 x 10(-6) mol.L-1 respectively. It is suggested that the cytoprotective effects of daurisoline and its isomers were mediated by blocking Ca2+ influx into cells. PMID- 11938961 TI - [The A1- and non A1-effects of N6-(5-hydroxy-2-pyridyl)-methyl-adenosine on rat vas deferens]. AB - N6-(5-hydroxy-2-pyridyl)-methyl-adenosine (HPMA) is a novel N6-substituted adenosine analogue recently obtained from Armillaria mellea (an edible fungus on which depends the growth of the famous Chinese traditional drug Gastrodia elata). It has been shown to have some characters of A1 receptor agonists of purinergic nerve. In this study, we compared the effects of HPMA with that of N6 Cyclohexyladenosine(CHA), an A1 selective agonist, on rat vas deferens in vitro, and found remarkable differences between them. In our study, HPMA dose dependently decreased the contraction responses to exogenous Phenylephrine (PE), Norepinephrine (NE) and Acetylcholine(ACh) on rat vas deferens, while CHA showed no effect on these responses. 8-Cyclopentyl-1,3-dipropylxanthine(DPCPX), an A1 selective antagonist, did not show any influence on these effects of HPMA, indicating that the depression effect of HPMA may be through a non-A1 mechanism. Using HPMA to decrease about 50% of the twitch responses evoked by field stimulation on rat vas deferens, the responsiveness to exogenous ACh seemed to be similar to that without HPMA pretreatment. These indicate that HPMA at this dosage (IC50 dosage) preferentially acted on presynapse (may be the A1 receptor) to attenuate the release of neurotransmitters. At a high dosage (10(-5) mol.L-1), HPMA abolished the neurogenic twitch responses evoked by electrical field stimulation, while the responsiveness of rat vas deferens to exogenous ACh was decreased showing both pre-synapse and post-synapse depression. PMID- 11938960 TI - [Protective effects of (-)-S.R-daurisoline on neuronal injury in rat primary cortical cultures]. AB - The neuroprotective effects of (-)-S.R-daurisoline on glutamate-induced neurotoxicity were studied in rat primary cortical cultures. The inhibitory effects of (-)-S.R-daurisoline on glutamate-elicited free intracellular Ca2+ increase were also studied in freshly dissociated single brain cells isolated from new born rat using AR-CM-MIC Cation Measurement System. Our experimental results demonstrated that (-)-S.R-daurisoline could obviously inhibit the neurotoxicity induced by glutamate and significantly increased cell viability in dose-dependent manner. In inhibiting glutamate-induced neurotoxicity, the IC50 value of (-)-S.R-daurisoline was found to be 3.4 mumol.L-1. (-)-S.R-daurisoline was also shown to markedly inhibit glutamate-elicited increase of cytosolic free Ca2+ concentration in dose-dependent manner with IC50 value of 2.0 mumol.L-1. Our results showed that (-)-S.R-daurisoline has an obvious protective effect on glutamate-induced neurotoxicity in primary cortical cultures. The protective mechanism of (-)-S.R-daurisoline may be relevant to inhibit Ca2+ influx into cells via glutamate-mediated ligand-gated ion channels. PMID- 11938962 TI - [The protective effects of nerve growth factor on peripheral nerve injury]. AB - The protective effects of nerve growth factor (NGF) on peripheral nerve injury and sensory neuron growth were studied in three models: peripheral sympathetic nerve injury induced by intraperitoneal injection of 6-hydrodopamine (6-OHDA), mechanically injured ulnar nerve and cultured dorsal root ganglion (DRG) of newborn rats in vitro. The results indicate that NGF dose-dependently increased norepinephrine content in mouse submandibular gland and cell numbers of C7 (seventh cervical vertebra), T1 (first thoracic vertebra) dorsal root ganglion in rabbits, and also dramatically promoted the growth of neuronal projections in cultured DRG. These results suggest that NGF has significant neuroprotective action in peripheral nerve injury and a strong neuronutrition on sensory neuron. PMID- 11938963 TI - [Effect of ginsenoside Rb1 on repeated stress-induced sexual deficiencies in male mice]. AB - The effect of ginsenoside Rb1 has been studied on sexual deficiencies induced by repeated hanging stress. Male mice were stressed by hanging once daily(9:00 am 2:00 pm) for 10 days(1-3 day hung for 2 h, 4-6 day hung for 3 h, 7-9 day hung for 4 h, 10-11 day hung for 5 h). On day 10, they were exposed to female mice treated with estradiol and progesterone and their sexual behaviors (licking, mounting, mating) were assessed at 7:00-9:00 pm. The repeated hanging stress was found to reduce sexual behaviors and decrease plasma testosterone level in mice. Treatments with ginsenoside Rb1(2.5, 5, 10 mg.kg-1, i.p.) 30 min before each stress prevented the repeated stress-induced sexual deficiencies and raised plasma testosterone level. The mechanism of the protective action of ginsenoside Rb1 may be attributed to its action in maintaining normal plasma testosterone level. PMID- 11938964 TI - [Comparative molecular field analysis(CoMFA) of allylamine and benzylamine antimycotics]. AB - "Active analog approach" has been employed to search the pharmacophoric conformation of the allylamine and benzylamine antimycotics. A local minimum energy conformation, which is very similar to the crystallographically determined coordinate of naftifine or terbinafine, has been applied to build all compounds. Comparative molecular field analysis (CoMFA) has been used to examine the correlations between the activities against 6 common human pathogenic fungi and the physicochemical properties of 62 allylamine and benzylamine compounds which had been aligned by rms fit rule. The predictive abilities of the resulting 3D QSAR models have been tested by 8 new synthetic compounds and 5 reported compounds. Finally, we propose an interaction pattern between allylamine, benzylamine antimycotics and the active site of pseudoreceptor based on the results of 2D-QSAR and 3D-QSAR. PMID- 11938965 TI - [Synthesis of primaquine analogues and their antimalarial activity in mice]. AB - Ten 4-methyl-5-substituted-phenoxy-6-methoxy-8-[(1-ethyl-4-amino) butylamino] quinolines (10a-j) have been synthesized and evaluated preliminarily for both suppressive and causal prophylactic antimalarial activities. The results of preliminary screening test showed that three of these compounds exhibited significant activity against Plasmodium yoelii in mice, among which 10c was 4-8 times as effective as primaquine. Moreover, 10c was superior to primaquine in suppressive test against Plasmodium berghei K173 strain in mice. On the basis of ED50 and ED90, 10c was about 2 times as active as primaquine. Most of other compounds showed similar activity and the minority of them exhibited lower activity compared with primaquine. PMID- 11938966 TI - [The structures elucidation of isodopharicin D and F]. AB - Two new compounds were isolated from Isodon pharicus (Prain) Murata. Their structures were determined to be 3 alpha, 11 beta, 13 alpha-trihydroxy-entkaur-16 en-15-one (1), named isodopharicin D, and 11 beta, 13 alpha, 15 alpha-trihydroxy entkaur-16-en-3 alpha-beta-D-glucoside (2), named isodopharicin F by chemical and spectral evidence. PMID- 11938967 TI - [Studies on urinary metabolites of HH07A, a derivative of hainanensine, in rats]. AB - The metabolism of HH07A in the rat has been investigated by GC-MS. After oral administration of HH07A, the rat urine was passed through a macroporous XAD-2 resin column and eluted with methanol. The methanol extract was hydrolyzed with glucuronidase, extracted with dichloromethane and concentrated for TMS derivatization with GC-MS. The mass spectra of HH07A and its four metabolites as well as their derivatives were presented. The structures of the metabolites were proposed and the in vivo metabolic pathway of HH07A was given in Figure 6. PMID- 11938969 TI - [Molecular genetic marker identification of traditional Chinese drug hippocampus]. AB - Ancient DNA technique was used to extract DNA from 5 species of Chinese traditional drugs Hippocampus. The 12S rRNA gene fragment and cytochrome b gene fragment were amplified from DNA extract using Polymerase Chain Reaction (PCR) method. Restriction Fragment Length Polymorphism (RFLP) analysis and DNA sequence analysis were performed. The RFLP analysis method can identify 2 species of Hippocampus. The molecular genetic markers produced by DNA sequence method can identify all 5 species of Hippocampus. This method will be valuable for the identification of other animal drugs. PMID- 11938968 TI - [The uptake characteristics of 5-fluorouracil in the Caco-2 model system]. AB - The uptake characteristics of 5-fluorouracil in the Caco-2 model system were studied. The uptake of 5-fluorouracil was determined at different pH and concentrations, and in the presence of various inhibitors. The results indicated that the uptake of 5-fluorouracil was the best at pH 6.0. The rate of uptake was saturable with a Km of 24 mmol.L-1, and a Vmax of 20.9 nmol.min-1.mg-1 protein. The uptake was inhibited by noncompetitive inhibitors such as NaCN, ouabain, and dipyridamole. The uptake was also inhibited competitively by analogous compounds such as uracil, thymine, and uridine (but not by hypoxanthine). In conclusion, the evidence suggests that 5-fluorouracil was transported by uracil carrier in Caco-2 cells. PMID- 11938970 TI - [Progress in studies on the artemisinin (qinghaosu)-type antimalarials]. PMID- 11938971 TI - [Effect of acrylonitrile on the activity of Ca(2+)-ATPase and phosphorylase A of liver in rats]. AB - The activity of Ca(2+)-ATPase and phosphorylase A(P-A) of liver in rats was determined to study the effect of acrylonitrile (ACN) on calcium homeostasis and to clarify the toxicological mechanism of ACN. Adult male Sprague-Dawley rats were administered ACN daily per os for 42 days, at the dosage of 0, 10, 30 and 50 mg.kg-1. The activities of Ca(2+)-ATPase and phosphorylase A was determined at 14, 28, 42 days after ACN administration. The results indicated that ACN could significantly inhibit the activity of Ca(2+)-ATPase and increase the activity of P-A (P < 0.01), especially in the high dose group and exposed for 42 days, and there was significantly dose- and time-respond relationship (P < 0.01). ACN contamination could result in the disharmony of calcium homeostasis of liver in rats. PMID- 11938972 TI - [Inhibition of glutamate uptake in rat brain synaptosome by methylmercury]. AB - The effect of methylmercury on the glutamate uptake in rat brain synaptosomes were studied in vitro by using a high affinity uptake test. The result showed that methylmercury(1 x 10(-9) -1 x 10(-4) mol/L) could decrease 3H-L-glutamate uptake in rat cerebral cortex, hippocampus, striatum and cerebellum synaptosome by 0.7%-74.4%, 7.5%-91.1%, 1.4%-76.6% and 4.4%-77.0%, respectively, in a dose dependent manner, and their IC50 were 7, 9, 3.6, 2.2 and 4.3 x 10(-6) mol/L, respectively. It suggested that methylmercury could obviously inhibit glutamate uptake in the synaptosomes of rat brain. PMID- 11938973 TI - [Changes of neurobehavioral functions in workers exposed to manganese]. AB - The aim of this study was to assess the neurotoxic effects associated with the level of airborne manganese in the work place. A cumulative exposure index(CEI) was calculated for each subject. The neurobehavioral test battery, in terms of choosing reaction time, finger tapping, digit symbol, stylus in hole, Benton visual retention and Santa Ana dexterity was applied in the study. The manganese exposed workers had slower motor and response speed, poorer visuomotor coordination and steadiness. There was a dose-response relationship tendency between the cumulative exposure index and the behavioral function. Hence the neurobehavioral test may be a sensitive method in detecting early changes in motor and cognitive function among manganese-exposed workers. The study also demonstrates that the cumulative exposure of manganese about 1 mg/m3 per year may induce subclinical signs of intoxication. PMID- 11938974 TI - [Lipid peroxide damage in retinal ganglion cells induced by microwave]. AB - The determination of lipid peroxide damage in the primary cultured pig retinal ganglion cells induced by microwave can provide some experience on the effect of microwave and the protection from its damage. Retinal ganglion cells were cultured in vitro and exposed to different intensities or time of microwave, and cultured for another 48 hours. The content of superoxide dismutase (SOD) and malondialdehyde (MDA) was detected. Results showed that the content of MDA was increased and SOD decreased after radiation. After 48 hours in culture, the content of MDA recovered, and SOD increased. It is concluded that microwave induced the lipid peroxide damage in primary cultured retinal ganglion cells. These damages were recoverable in certain range of intensity or time. PMID- 11938975 TI - [Effect of cadmium on the beta-adrenoreceptor of splenic cell of rats and its immunotoxicity]. AB - Rats were given cadmium(Cd) alone or Cd with beta-antagonist in order to study the effect of Cd on beta-adrenoreceptor density of rat splenic cells. The subsets of T-lymphocytes (CD4+, CD8+, CD4+/CD8+) and the proliferation of T lymphocytes were also detected. Cadmium alone stimulated and increased beta-adrenoreceptor density of rat splenic cell membrane. The immunotoxicity of Cd showed that the proliferation of T-lymphocytes was inhibited and the subsets of T cells were changed. But in the animals with beta-antagonist, the immunotoxicity of Cd on T cells was decreased. It is concluded that beta-adrenoreceptor may play an important role in the mechanism of cadmium on cell immunotoxicity. PMID- 11938976 TI - [Effects of alcohol and metabolite acetaldehyde on the proliferation of astroglial cells of fetal brain]. AB - To explore the mechanism of the retardation and abnormality of nerve system development induced by drinking alcohol during pregnancy, 3H-TdR incorporation was used to study the proliferation of astroglial cells of fetal brain induced by alcohol and its metabolite acetaldehyde in vitro. The results showed that alcohol and acetaldehyde obviously restrain the proliferation of astrocytes. The effect of restrain induced by acetaldehyde is stronger than that by alcohol. Nerve system development retardation and abnormality resulted from alcohol is likely to be the restrain of proliferation induced by alcohol and its metabolite acetaldehyde. PMID- 11938977 TI - [Risk state analysis on lung cancer of male in Xuanwei, China]. AB - The analysis of lung cancer risk factors among male population was based on the data of a retrospective cohort study conducted during 1976-1996 in Xuanwei, China. The results verified that the indoor air pollution from burning smoky coal was the main cause of high lung cancer mortality, and found that cigarette smoking was also significantly related with the lung cancer mortality. Using risk state analysis theory, fuel types(including household stove improvement), smoking and the history of chronic bronchitis were selected as the covariance for the analysis. A risk state analysis model for male lung cancer in Xuanwei was established. The model can be used to quantitatively estimate the risk of lung cancer and to determine high risk population and individuals. The model is helpful for the prevention of lung cancer in Xuanwei. PMID- 11938978 TI - [Effect of fluoride exposure on synaptic structure of brain areas related to learning-memory in mice]. AB - The learning-memory behavior was tested in mice on a Y-maze after drinking different concentration of sodium fluoride. The impairment on the structure of Gray 1 synaptic interface in the CA3 area of mice hippocampus were quantitatively analyzed by electron microscopy and computer image processing appliance. The main results are as follows: the learning ability of mice drinking high concentration of fluoride presented remarkable deterioration, the thickness of post-synaptic density (PSD) was decreased, and the width of synaptic cleft was remarkably increased. The results suggested that the impairment on the learning capability induced by fluorosis may be closely related with the pathological changes of synaptic structure in the brain of mice. PMID- 11938979 TI - [Effect of manganese on the brain extrapyramidal development of rat offspring]. AB - The effect of manganese (Mn) exposure on the behavior and extrapyramidal system of offspring were studied. Pregnant rats and their offspring in the experimental group were exposed to manganese from drinking water containing 2 g/L (low Mn exposed group) or 10 g/L (high Mn-exposed group) of MnCl2 respectively. The latent period for running from the center field to any arm of Elevated Plus Arm Maze of both low and high Mn-exposed group was significantly reduced. But the latent period for reaching the platform above the water in Morris Water Maze of the Mn-exposed group was not different from that of control group. There was a significant increase in the areas, glial fibrillary acid protein(GFAP) immunoreactivity and the average proportional densities of GFAP-positive elements in nucleus caudate and accumbens in the high Mn-exposed group, and a significant reduction in the tyrosine hydroxylase immunoreactivity (TH) and average proportional densities of TH-positive elements in substantia nigra, ventral tegmentum area of midbrain and nucleus caudate in the high Mn-exposed group. PMID- 11938980 TI - [Animal model for Alzheimer's disease based on the blockade of neuronal nicotinic acetylcholine receptors]. AB - The intrahippocampus injection of Kappa-bungarotoxin (K-BGT) caused a significant impairment in learning and memory abilities of mice, in order to create an animal model for Alzheimer's disease. The spatial learning and memory abilities in the Morris water maze in the model group (K-BGT impaired mice) decreased significantly compared with control group. K-BGT impaired mice also showed learning and memory reduction with shorter latency and more mistakes in step through test. PMID- 11938981 TI - [Time-effect relationship between the positional microinjection of HCY-2 gene and the neural tube teratogenesis of chick embryos]. AB - The time-effect relationship between positional transferred novel gene(HCY-2) and neural tube teratogenesis, and their possible mechanisms were studied. An eukaryotic expressing vector which containing whole-length HCY-2 cDNA was microinjected into chick embryos in culture at days 0, 1 and 2 (approximately Hamburger-Hamilton stages 1, 6 and 12) mediated by lipofect AMINE reagent. The techniques of RT-PCR, immunohistochemical staining, scanning electron microscope(SEM) and transmission electron microscope (TEM) were used to investigate the expression and distribution of HCY-2 mRNA with its coding product and dysmorphogenesis of the neural tube at 96 h (approximately stage 22). Neural tube defects (NTDs) were discovered in every transferred gene group, only the day 1 embryos which positional site injected was the area pellucida of head, however, the rate of NTDs was the highest (35.3%). There was an obvious time-effect relationship. The phenotypes of NTD were encephalocele, anencephaly, spina bifida and microcephaly. The embryos with transferred gene could express HCY-2 mRNA and its coding product, and the HCY-2 protein mainly distributed in embryonic brain cells as compared to controls. It was found that HCY-2 gene could result in abnormal ultrastructure at the surface and inside of cells under SEM and TEM. It has been observed apoptosis at the sites with NTDs. It is concluded that HCY-2 gene may be a new genotoxic factor, which plays an important role in the mechanisms of neural tube teratogenesis during the early developing stage of embryos. PMID- 11938982 TI - [Vegetables rich in carotenoids on the vitamin A status of children]. AB - Vitamin A (VA) deficiency is still an important nutritional problem in our country. In order to determine whether plant carotenoids ingestion can improve VA nutrition, a study by using yellow and dark green vegetables was conducted in two classes of a kindergarten for ten weeks in September through December in 1996. The VA nutritional status was marginal to adequate among 41 children (5.3-6.6 year-old), and serum retinols in 39% of these children were below 0.30 mg/L. Each child in class A was provided with about 238 g/d of green and yellow vegetables (spinach, Chinese chive, carrots, and red yams) and 34 g/d of light colored vegetables (cabbage, Chinese cabbage, potato, cucumber, turnip and winter melon). Each child in class B consumed the usual diet with only 56 g/d of green and yellow vegetables and 193 g/d of light colored vegetables. Serum retinol concentration collected before and after the intervention were used to assess VA nutritional status. The results showed that vitamin A nutrition was improved by increasing the intake of green and yellow vegetables. Serum retinol was sustained in the group fed green and yellow vegetables and decreased in the group fed light colored vegetables. Thus, dietary green and yellow vegetables could provide adequate VA nutrition in these children. PMID- 11938983 TI - [Effect of physiological dosage of estrogen on the prevention of bone loss in rats induced by ovariectomy]. AB - The effect of small dosage of 17 beta-estradiol (E2) on bone metabolism, bone mineral density (BMD) and biomechanical properties of bone in ovariectomized rats were observed. Twenty-one rats were divided into three groups. They were sham operated group (A), ovariectomized group (B), and ovariectomized and treated with 17 beta-estradiol group(C). Rats were sacrificed after 9 weeks of treatment. Serum alkaline phosphatase (ALP) and osteocalcin(OC) contents were significantly increased; bone mineral content (BMC) of right femur and the third lumbar body(L3) and the biomechanical properties of right femur were decreased in group B. Treatment with E2 can reverse the changes of BMD and BMC in ovariectomized rats. There was no significant improvement on biomechanical parameters in group C rats. It is concluded that physiological dosage of E2 may inhibit bone resorption and bone formation and increase bone mass, but it may not improve the strength of bone. PMID- 11938984 TI - [Effect of calcium and cristata L extracts on bone mineral density and urinary mineral of rats]. AB - In order to study the effect of cristata L for preventing fluoride toxicity, forty male SD rats were randomly divided into five groups. The control group(NC) was fed with staple feed stuff and tap water, the other four groups were fed with staple feed-stuff and high fluorine water. Except the high fluorine group(F), the other groups were fed with cristata L(FC), high calcium(FCa) and cristata L with high calcium(FCCa) respectively. The results showed that the serum calcium, urinary sodium, calcium, hydroxyproline(HOP) and fluorine contents of high fluorine groups were higher than that of control group; and the urinary calcium content of high calcium group was higher than that of control group. The body weight, BMC and BMD of high fluorine groups was lower than that of control group. The weight of groups FC, FCa and FCCa were significantly higher than that of group F. The BMC and BMD of group FCCa were significantly higher than that of group F. This implies that the food high in fluorine can affect the bone mineral density and urinary minerals of rats, the food supplemented with calcium can reduce the effect of high fluorine, and the food supplemented with both calcium and cristata L is better. PMID- 11938985 TI - [Sequencing and analysis of 16S rDNA sequences for P. cocovenenans subsp farinofermentans]. AB - In order to fix the phylogenetic taxonomic position of P. cocovenenans subsp. farinofermentans and describe its relationships with other closed species, seven primers were designed to amplify and sequence the 16S rDNA of 4 strains of it by using a clonal sequencing or direct sequencing method. The 16S rDNA sequence of the 4 strains were aligned with the 16S rDNA sequences of other species in genus Burkholderia, and the polygenetic tree was produced by using a Clustal V and treeview software. The results showed that P. cocovenenans subsp. farinofermentans was in a high homology with Burkholderia gladioli and Burkholderia cocovenenans, and they formed an independent phylogenetic clustal of genus Burkholderia. PMID- 11938986 TI - [Effects of vitamin A deficiency on the development and growth of rat embryos]. AB - Effects of vitamin A deficiency on the development and growth of rat embryos were studied. Serious vitamin A deficiency markedly reduce body weight, body length and tail length, and induce incomplete development of skeleton and induce pathological change of brain and kidney of rat fetus. It was concluded that the development and growth of the embryos was affected and the brain and the kidney of fetus were damaged in serious vitamin A deficient rats. PMID- 11938987 TI - [Studies on immunoassay for detecting fumonisin B1]. AB - Direct competitive inhibition enzyme-linked immunosorbent assay for determination of fumonisin B1 (FB1) in corns were developed by using anti-FB1 monoclonal antibody. The minimum detectable concentration of FB1 was 10 ng/ml, and the standard curve was linear between 10 ng/ml and 5 micrograms/ml. The FB1 concentration of some corn samples from Xinjiang, Anhui, Hilongjiang provinces and Haerbin city were detected. The result imply that 34 out of 156 samples were positive for FB1. The concentrations were in the range of 0.18-31.32 micrograms/g. The average concentration was 12.04 micrograms/g. PMID- 11938988 TI - [Studies on the preparation and characters of anti-AFB1 antibody from the yolk of laying hens]. AB - AFB1O-BSA conjugates were injected into four laying hens (A, B, C and D). The yolk anti-AFB1 antibodies A,C and D produced were obviously increased after 90 days of the first injection, and reached the peak after 135 days, and started dropping after 165 days. The indirect ELISA titer of yolk antibodies A,C and D were 1:8000, 1:6000 and 1:6000 respectively. The specificity of the antibody A was good, but a cross-reaction with AFB2, AFG1, AFG2, with an exception of AFM1, was found. The sensitivities of AFB1, AFB2, AFG1 and AFG2 were 6, 25, 125 and 2495 micrograms/L respectively. PMID- 11938989 TI - [Pathogenic gene analysis of Listeria isolated from foods]. AB - In 1996-1997, Listeria in foods from 12 provinces of China were detected by using the national standard examination method GB 4789.30-94 (routine traditional methods). Twenty four strains Listeria monocytogenes and 141 strains Listeria were identified by biochemical procedures and animal toxicity experiment. Polymerase Chain Reaction(PCR) was used to amplify two specific DNA fragments(719 bp and 446 bp) of listeriolysin O and internalin genes with two pairs of primer- Hly1-2 and Inl1-2, respectively. The experiment results showed that 57 strains contained only internalin, 27 strains contained both internalin and listeriolysin O from the pathogenic genes of isolated strains. PMID- 11938991 TI - [Ecological risk assessment and its application]. AB - Ecological risk assessment, closely linked to environmental toxicology, is becoming one of the most widely expanding disciplines in environmental science. A number of frameworks for ecological risk assessment have been developed by the USEPA, Environment Canada and European Commission in recent years. This review described the common elements, the process and steps of ecological risk assessment. In addition to drawing upon the recent published literature, we have developed some examples of applications based on our collective experiences in conducting ecological risk assessments. PMID- 11938990 TI - [Passive personal sampler for exposure to low concentration of volatile organic compounds]. AB - A passive personal sampler has been developed for monitoring volatile organic compounds(VOCs) in ambient air and nonoccupational exposure. The sampler collecting target VOCs is based upon the mechanism of molecular diffusion and absorption onto a single layer of active carbon fibre (ACF) absorbent. After exposure, the ACF is transferred from the sampler into stainless steel tubes for thermal desorption. The desorbed target VOCs is analyzed by capillary GC. Under the condition of facing velocity of 10-260 cm/s, relative humidity of 30%-80%, and temperature of 10-30 C, the sampling rate for benzene, toluene, tetrachloroethylene, p-xylene and styrene is 25.74, 25.16, 25.85, 8.16 and 6.47 ml/min respectively. The analysis on the new passive sampler demonstrated that its relative overall uncertainty for target VOCs was in the range of +/- 25% compared with the active ACF tube sampling method. PMID- 11938992 TI - [Development of air microbe sampler with micropore filter membrane]. AB - Dual head air microbe sampler with a large flow capacity of 100 liters per minute was manufactured. The sampler possesses simple structures, light weight, convenient operation and higher sampling efficiency, equal to 77%-96% of the grade six Anderson sampler. Sampling time is a main factor affecting sampling efficiency. This device is suitable for the determination of degree of biological clean air in clean environment and the detection of the pathogenic microbes in ward and public places and also microbes in ordinary environment. PMID- 11938993 TI - [Study on the contamination of algae and microcystins in water sources of Zhengzhou City]. AB - Planktonic algae and microcystins (MCYST) were detected constantly in water sources of Zhengzhou City from 1996 to 1997. There is significant seasonal variation of the density of algae. The peak of algae occurring in February and December is about 1,068,000/L. The contents of MCYST in July, 1995 and in January, 1996 were over 200 ng/L. According to the Carlson's trophic state index(TSI), and indexes of total nitrogen, phosphorus, COD value and algae density, an eutrophication of water source having emerged in Zhengzhou is considered. PMID- 11938995 TI - [Investigation on the contamination of Listeria monocytogenes in seven kinds of foods]. AB - An examination on 3746 samples of 7 kinds of foods from 12 provinces of China for the detection of Listeria monocytogenes (LM) was carried out in 1996-1997. The detected rates of LM in commercial cold drinks, raw meat, sterilized milk products, cooked meat products, raw milk and aquatic products were 1.39%, 1.53%, 0.52%, 0.47%, 0.72% and 0.19% respectively. The result showed that LM present in the contaminated foods in a potential hazardous factor of foodborne disease. PMID- 11938994 TI - [Effect of deoxynivalenol and aflatoxin G1 on apoptosis of human blood lymphocytes in vitro]. AB - Effect of deoxynivalenol (DON) and aflatoxin G1 (AFG1) on human blood lymphocytes in culture were studied with flow cytometric DNA content analyses and DNA agarose gel electrophoresis. DON and AFG1 are two of the predominant mycotoxins contaminating foodstuffs or residents in a high incidence of esophageal cancer in Cixian County, Hebei province. A typical sub-diploid apoptosis peak was demonstrated in lymphocytes treated with DON and AFG1. A significant dose-effect response and time-effect correlation could be found between apoptosis rates and mycotoxin concentrations (DON: 50-2000 micrograms/L and AFG1: 3.12-2000 micrograms/L) and the treated time (DON: 2-72 hours and AFG1: 2-24 hours). The lymphocytes treated with DON and AFG1 showed characteristic 'ladder' pattern in agarose gel electrophoresis. All results confirmed that DON and AFG1 could induce and accelerate apoptosis in human peripheral blood lymphocytes. PMID- 11938996 TI - [The antioxidant activity of wild jujubi, crataegus and grape in vitro]. AB - Two oxidative damaging systems, Fe(2+)-Cys and tert-butyl hydroperoxide, were used to induce the production of malondialdehyde (MDA) in rat liver microsome and erythrocytes. The inhibition rate of wild jujubi, crataegus and grape juice at the concentration of 2.5-3.3 mg/ml to Fe(2+)-Cys system on the production of MDA were 46.2%, 98.3% and 99.1% respectively. The inhibition rate of jujubi (8.3 mg/ml), crataegus (13.9 mg/ml) and grape (55.6 mg/ml) to tert-butyl hydroperoxide were 38.7%, 38.7% and 58.5% respectively. The capability of scavenging O2.- and .OH was measured by ESR technique. It was found that the capability of these juices to eliminate O2.- and .OH generated by the xanthine/xanthine oxidase system and H2O2-FeSO4 system was strong. The elimination rate of jujubi (50 mg/ml), crataegus (50 mg/ml) and grape (134 mg/ml) to O2.- were 84.0%, 85.8% and 74.7% respectively and the rate to .OH were 96.7%, 97.8% and 86.3% respectively. The results indicated that these three fruits bear antioxidant activity. PMID- 11938997 TI - [Research on the mechanism of the fatty acid composition on the tumorigenic danger induced by chemical tumorigenic material]. AB - Rats were given 5 kinds of diets which contained 15% (w/w) fat and different fatty acids composition. The rats were given methyl-nitrosurea (MNU) to induce colonic tumor. Proliferation cell nuclear antigen (PCNA), cell kinetics, membrane fluidity, the activity of alkaline phosphatase (ALP) and the content of prostaglandin E2(PGE2) in colonic mucosa were determined in order to assess the fatty acids composition on the colonic cell tumorigenesis. The results indicated that the cells of PCNA, cells of PI labeled in S period and the activity of ALP were the highest in the 3rd group which contained lowest saturated fatty acids (SFA) and monounsaturated fatty acids (MUFA) and highest n-6 polyunsaturated fatty acids. Whereas, these indexes and PGE2 were lowest and membrane fluidity was the best in the 4th group which contained the highest 1-3 PUFA. It is considered that the inhibition tumorigenesis of n-3 PUFA may be related to its effects of decreaing PCNA, PGE2, cells in S period and increasing membrane fluidity. PMID- 11938998 TI - [The protective effects of total flavonoids from Lycium Barbarum L. on lipid peroxidation of liver mitochondria and red blood cell in rats]. AB - The protective effects of total flavonoids from Lycium Barbarum L. (TFL) on lipid peroxidation in mitochondria and red blood cells (RBC) induced by oxygen radicals produced by Fe2+ cysteine system were investigated. The mitochondria lipid peroxidation (measured as malondialdehyde, MDA) was significantly inhibited by TFL with a dose-response relation between the concentrations of 0.025 and 2.0 mg/ml, and the fluidity of mitochondria membrane was also protected effectively. It was observed by scan electron microscope, that the shape of RBC in the Fe2+ system was damaged significantly. The shape of RBC was remained with the addition of TFL. PMID- 11938999 TI - [Study on standards for safe and health-protective zone in firework plant]. AB - A retrospective investigation on technology and situation in the production of fireworks, the cause and hazard consequences of accidents in blossom firework enterprises was carried out. The risk factors and their origins, the potential effects on surrounding environments and residents, the manufacture processes producing special potential energy in these enterprises were summarized and assessed. In addition, the consequences of explosive fire accidents were assessed retrospectively by the principle of explosion mechanics and Hopkinson Scaling Law. The safe and health-protective zone of the blossom firework plant was suggested. PMID- 11939000 TI - [Research on the protective effect of wine against cardiovascular diseases]. AB - Recent studies reveal that wine is rich in phytoalexin and polyphenolic compounds, which can inhibit atherosclerosis by modulating metabolism of plasma lipid and lipoprotein, preventing platelet from aggregation and relaxing cardiovascular smooth muscle, etc.. A long-term moderately intake of wine can reduce the morbidity and mortality of ischemic heart diseases. PMID- 11939001 TI - [The use of immunological methods for the detection of phycotoxin and shellfish toxin]. AB - Harmful algae can do great harm to organisms even to human through food chain by accumulating in shellfish and fish or by themselves. The immunological examination technique is fast, easy and accurate even in trace amount of phycotoxin and shellfish toxin. The theory of immunological diagnosis as well as the prospect and defect of these methods and their application in phycotoxin and shellfish toxin determination are introduced. The method deserves spreading and using in routine monitoring program. PMID- 11939002 TI - [Healthy life years(HeaLY): a new indicator for measuring the burden of diseases]. AB - Healthy life years(HeaLY) is a new indicator for measuring the burden of diseases. The data of Beijing MONICA were analyzed and reevaluated by using this indicator, and were compared with another indicator, disability adjusted life years (DALY), for the burden of diseases provided by WHO and World Bank several years ago. PMID- 11939003 TI - [Effect of lead on the expression of immediate early genes in different regions of rat brain]. AB - In order to study the role of immediate early genes (IEG) in the neurotoxic mechanism of lead, expression of c-fos and c-jun genes in different regions of rat brain treated with lead (13 and 130 mg/kg) were observed by using immuno histochemical method. The observation and image analysis showed that after rats treated by lead for 2 hours, the c-fos and c-jun expression in cortex, CA3 area of hippocampus and cerebellum of rats were higher than those of control rats significantly (P < 0.05). The results indicated that IEG, as a transcription regular factor, may participate in the neurological toxicity damaging the learning and memory ability induced by lead. The study provided experimental basis for revealing the molecular neurotoxic mechanism of lead. PMID- 11939004 TI - [Study on the correlation of silicosis with antioxidant and antioxidase]. AB - Plasma vitamin C(P-VC), vitamin E(P-VE) and beta-carotene(P-beta-CAR) contents and the activities of superoxide dismutase(E-SOD), catalase(E-CAT) and glutathione peroxidase (E-GSH-Px) in erythrocyte in 73 silicosis patients and 60 healthy control subjects were measured. The average levels of P-VC, P-VE, P-beta CAR, E-SOD, E-CAT and E-GSH-Px of patients were significantly lower than those of the controls (P < 0.001). All indexes were correlated to the course, condition and pulmonary function of silicosis patients. The results analyzed by stepwise regression showed that the correlation between course, condition and pulmonary function of patients and P-VE and E-SOD was close. The balance between oxidation and the antioxidation in silicosis patients may be disturbed, and oxygen free radical reaction may be pathologically exacerbated. PMID- 11939006 TI - [The toxicity of combination of selenium, fluoride and arsenic on rat embryos]. AB - Whole embryo rotated culture technique was used to investigate the toxicity of combination of selenium, fluoride and arsenic on rat embryos at day 9.5 of gestation. The result of factorial analysis (3 x 3 x 3) showed that the main effect of combination of selenium, fluoride and arsenic on the developmental toxicity was synergistic. The mixtures with different level of these three chemicals in combination could result in different developmental toxicity. The low level combinations mainly caused teratogenic effect, and the high level combinations(selenium 2.0 micrograms + fluoride 10 micrograms + arsenic 1.0 microgram/ml culture media) caused lethal effect. The results suggested that the disorders of yolk-sac placenta in structure and function were one of teratogenic mechanisms for the combination of selenium, fluoride and arsenic. PMID- 11939005 TI - [Effect of dimethoate on the expression of c-fos gene in skeletal muscle]. AB - To elucidate the effect of organophosphate pesticides on the expression of immediate early gene in skeletal muscle, the concentrations of c-fos mRNA and protein were measured by RT-PCR and western dot blot techniques. The result showed that c-fos mRNA and protein were significantly increased in skeletal muscle of rat after dosing with dimethoate. These results indicated that c-fos might act as transcription factor and regulate other gene expression during the early period of organophosphate intoxication. PMID- 11939007 TI - [Apolipoprotein E polymorphism and its association with serum lipid level in Uygur nationalities from Xinjiang]. AB - To study the relative frequency of apoE allele and the influence of polymorphism on serum lipid and lipoprotein concentration in Uygur nationalities, apoE phenotype was analysed by polymerase chain reaction and restriction fragment length polymorphism (PCR-RFLP) in 163 Uygurs and 150 Hans. Serum lipid levels were measured in 101 Uygurs and 140 Hans of them. The results showed that the frequency of E4 allele was higher in the Uygurs than in the Hans. Total cholesterol and low-density lipoprotein cholesterol were higher in the two nationalities who bear E 4 allele. It is concluded that the distribution of apoE genotypes and allele-frequencies was significantly different, but the trent for the relationship between apoE phenotype and lipid and lipoproteins was similar in the two nationalities. PMID- 11939009 TI - [Preheating decrease the sensitivity of K562 cell to chemotherapeutic drugs]. AB - Hyperthermia is a powerful tool for enhancing the effect of many chemical drugs on killing cells, but too many times of hyperthermia could decrease the effect and increase the ability of tumor cell against chemical drugs. Heat stress protein 70(HSP70) mRNA of K562 cell was analyzed by RT-PCR, and the chemosensitivity of K562 cell was tested by MTT method before and after heat exposure. The results showed that HSP70 mRNA increased greatly after K562 cells exposed to 40 degrees C for 30 min and then it went to the top after 120 min heat exposure. The sensitivity of K562 cell to adriamycin (ADM) and cyclophosphamide (CTX) in a usual dosage (1/250 of one dosage in clinical use) was in middle degree before exposed to heat, and the sensitivity dramatically went down after K562 cell being heat-stressed. The sensitivity of heat-stressed tumor cells to chemotherapeutic drugs was decreased, which may come from the increased expression of stress proteins in cells. PMID- 11939008 TI - [Study on the risk factors influencing the epidemicity of epidemic cerebrospinal meningitis]. AB - Based on a 8-year surveillance data, a factor analysis theory was used to analyze the 12 risk factors influencing the epidemicity of epidemic cerebrospinal meningitis. The results showed that the first sixth risk factors were the rate of carrying meningococci A and B in the population, relative air humidity in January, morbidity rate in January, vaccination rate before epidemic, and the titre of antibody. Although the incidences of epidemic cerebrospinal meningitis are scattered, the risk factors influencing the epidemicity did not disappear. It is important to use meningococci A vaccine continuously and widely to strengthen the surveillance among migrated population and population at high risk. PMID- 11939010 TI - [Detection of apoptotic cells by comet assay]. AB - Comet assay is a method measuring DNA strand beaks on single cell level, which has been developed for about 10 years. Theoretically, apoptotic cells with extensive DNA internucleosomal fragments can be detected by this method. On the basis that apoptosis in immature mice thymocytes could be induced by dexamethasone(DEX), the detection of apoptotic cells by comet assay was studied and compared with agarose gel electrophoresis, morphological observations with Giemsa staining and TUNEL assay. Apoptotic cells showed a comet feature with a very small head, and a large and wide tail. The optimal conditions for electrophoresis in comet assay was 20 volt in 15 minutes. Comet assay is sensitive, valid, rapid and simple and is highly correlated with Giemsa staining and TUNEL assay in detecting apoptotic cells. PMID- 11939011 TI - [Effect of low dose gamma-irradiation and cigarette smoking on growth and reflex behavior in mice]. AB - Pregnant mice were exposed to different level of gamma-irradiation(60Co 0, 0.25, 0.5 and 1.0 Gy) and water solutions containing different contents of substance from cigarette smoke(0, 0.5, 1.0, 1.5 and 2.0 cigarette/mouse). Results showed that the effect of low dose gamma-irradiation and high concentration cigarette smoke substance could delay the appearance of four physiological markers (eye opening, pinna detachment, hair growth and tooth growth) and the acquisition of four reflexes(surface righting, air righting, virual placing and auditory orienting). A significant combined effect was also observed. PMID- 11939012 TI - [Distribution of lead in fetal rat tissue at different days of pregnancy]. AB - In order to reveal the mechanism of the transplacental transfer of lead from mothers to fetus, tissues lead content of fetal rat at different days of pregnancy were determined and compared. Lead was administered to four groups of 4 week-old female Wistar rats for 50 days. The doses of lead were 0, 10, 50 and 200 mg/L in drinking water respectively. Rats were mated with non-lead-treated male rats. The pregnant rats in each group were then divided into four sub-groups by random. The pregnant rats in four sub-group of each group were sacrificed in turn at 10, 14, 17 and 20 days of gestation (dg). Blood lead of pregnant rats, lead burden of whole fetus, lead concentration of fetal brain, fetal liver and placenta were determined. Results indicated that lead concentration in fetal brain kept unchanged during the period of gestation, but lead content in whole fetus and fetal brain were increased with the days of gestation. But the lead concentration in fetus at 10 dg was higher than that at 14 dg. It seemed that higher lead absorption by the fetus was related to the newly emerged lead bonding site in fetus, especially during the calcification of bones. PMID- 11939013 TI - [Study on the hepatic toxicity of cerous nitrate in rats]. AB - Rats were injected with a single dose of cerous nitrate Ce (NO3)3 (150 mg/kg) intra-peritoneally and killed at 3, 6, 12, 24 and 48 hours later. The results showed that the concentrations of protein and malondialdehyde (MDA) in liver increased, but the concentration of glutathione (GSH) and the activity of superoxide dismutase (SOD), catalase (CAT), glutathione peroxidase (GSH-Px) and glutathione sulfatransferase (GSH-ST) decreased after Ce3+ administration. The results suggest that lipid peroxidation in liver may be an early consequence of Ce3+ exposure and the decrease of GSH might be considered as the cause of lipid peroxidation. PMID- 11939014 TI - [Study on the relationship between the structure of chlorobenzenes and their toxicity to guppy]. AB - The quantum chemical indexes of 12 chlorobenzenes were calculated by CNDO/2 method. Quantitative structure-activity relationship between the quantum chemical indexes of chlorobenzenes and their toxicity to Guppy was calculated by step regression analysis. The linear correlation equation is as follows: DLC50 = -0.15 + 6.865 sigma QR, n = 12, R = 0.9867 s = 0.15, F = 367.64. Results showed that the toxicity of chlorobenzenes to the aquatic Guppy are increased as the increase of their ability to accept electrons. PMID- 11939015 TI - [Malignant transformation of HPV-18 immortalized human bronchial epithelial cells induced by tobacco-specific nitrosamine NNK]. AB - The malignant transformation of HPV-18 immortalized human bronchial epithelial cell(BEP2D) induced by tobacco-specific nitrosamine NNK and its biological characteristics were investigated. BEP2D cells treated with NNK at the concentration of 500 micrograms/ml for 24 hours could be subcultured continuously in vitro. The 15th passage cells in soft agar (0.038%) increased 11 fold compared with that of control cells (0.0033%). The 25th passage cells could grow into tumor in nude mice. The tumor was a squamous cell carcinoma in morphology confirmed by histopathological examination. The results indicated that the malignant transformation induced by NNK was a sequential process. This culture system provides a potential tool for the study of cell and molecular mechanism in the multistage carcinogenesis of human bronchial epithelial cells. PMID- 11939016 TI - [Effect of antioxidant vitamins on lipid peroxide injury of aortic endothelial cells induced by oxidatively modified low-density lipoprotein in vitro]. AB - The effect of antioxidant vitamins including vitamin E, vitamin C and beta carotene on lipid peroxidation injury of aortic endothelial cells induced by oxidatively modified low density lipoprotein (oxLDL) were observed. Bovine aortic endothelial cells were incubated in culture media with antioxidant vitamins for 12 hours, and then with oxLDL (0.1 g/L) for 24 hours respectively. The results showed that the malondialdehyde (MDA) content in culture media of the groups with oxLDL was significantly higher than those of groups with vitamin E, vitamin C and beta-carotene alone. The number of HL60 monocyte cells adhered to bovine aortic endothelial cells was obviously higher in oxLDL groups than those in groups with only antioxidant vitamins. This study indicated that antioxidant vitamins are protective on vascular from injury induced by oxLDL, including decreasing lipid peroxidation and reducing adherence of monocyte cells on endothelia and seemed to be promising in the prevention of atherosclerosis. PMID- 11939017 TI - [Heat strain for personnel wearing three levels of protective ensembles at moderate and heavy workloads]. AB - The effects of individual protective ensembles and workloads on personnel's physiological heat strain were experienced by personnel in a hot environment (35 degrees C, 55% relative humidity). Six young males performed treadmill tests at both moderate (ML: 1.11 m/s, 5% grade) and heavy (HL: 1.34 m/s, 5% grade) workloads. They wore three levels of protective ensembles: protective mask only (PM), protective clothing only (PC), and full protective ensembles (FP). Tests lasted 60 min and 45 min, respectively, for ML and HL. Physiological measurements included heart rate (HR), rectal (Tre) and skin (Tsk) temperature, sweat rate (SR), and sweat evaporative rate (ER). Differences between the ensembles and the workloads were significant for HR responses. The increase rate in Tre and Tsk, and SR, ER were significantly reduced from PM and PC to FP condition during ML and Hl tests. The sweat evaporation restriction of ensembles and the increase of heat production from exercise are the main factors of enhancing the heat strain. PMID- 11939018 TI - [Assessment for effect of low level lead-exposure on neurobehavior in workers of printing house]. AB - WHO Neurobehavioral Core Test Battery (NCTB) was conducted among 28 lead-exposed workers (mean age 24.84, SD2.85) in printing house and 46 controls (mean age 22.78, SD1.45), in order to assess whether low level lead exposure may be related to neurobehavioral dysfunction. The items of test were: 1. Profile of mood state(POMS), (2) Simple reaction time, (3) Digit span, (4) Santa Anna manual dexterity, (5) Digit simbol, (6) Benton visual retention; and Prusuit aiming test. In all the NCTB test values, there was no significant difference between two groups. Multiple stepwise regression analysis shows that exposure duration is related to neurobehavior scores. Mild lead exposure may affect neurobehavior in some degree but not significant. PMID- 11939019 TI - [Effect of impactor sampling on the ratio of respirable dust concentration to total dust concentration]. AB - The aim of this study was to verify the change of the ratio of Respirable Dust Concentration (RDC) to Total Dust Concentration (TDC) taken by two-stage impactor sampling. The results showed that there was a correlation between the ratio of RDC/TDC and TDC (correlation coefficient was 0.8576, 0.8689, 0.8736 and 0.9674, respectively) under the condition of identical sampling volume, and the ratio increases along with the elevated TDC. PMID- 11939020 TI - [Homocysteine-induced neural tube defects in chick embryos and protection of folic acid]. AB - To reveal whether homocysteine(HCY) will be able to induce neural tube defects (NTDs) and possible teratogenic mechanisms, and identify the interventional effects of folic acid and vitamin B12 on HCY-induced NTDs, 1274 chick embryos treated with L. D-HCY (0-16 mumol/embryo) were detected by the methods of teratogenic trial, scanning electron microscopy(SEM), Nile blue sulfate vital staining, TdT-mediated dUTP nick end labeling(TUNEL) methylgreen-pyronine special staining for showing nucleic acid, and interventional assay of folic acid and VB12 in different developmental stage. Results showed that HCY could induce teratogenesis in chick neurulation and organogenetic period, in dose-response relationship (P < 0.0001). The main forms of NTDs were exencephaly, cranioschisis and spina bifida. It has been first found that HCY could target excessive apoptosis of cells of embryonic nervous system, which consists with the sites of embryos developing NTDs. HCY could inhibit chick yolk sac vessel formation and blood circulation, and damage amniotic membrane. Under SEM it was observed that HCY resulted in abnormal ultrastructures, for example cavernous damages, atrophied microvilli and so on. After the injection of 5 micrograms/embryo folic acid, the teratogenicity of HCY (8 mumol/embryo) was significantly antagonized, the occurrence rate of NTDs was down from 43.5% to 0(P < 0.05). However, vitamin B12 (1 microgram/embryo) did not obviously attenuate the teratogenicity of HCY in day 6 chick embryos. These data prove that HCY per se causes dymorphogenesis of the neural tube. Apoptosis may play an important role in the etiopathology of NTDs. Folic acid can effectively prevent from the homocysteine-induced NTDS. Causes of homocysteine-induced NTDs may result from interaction or combination of different mechanisms. PMID- 11939021 TI - [Study on toxicity of P-Dichlorobenzene]. AB - The authors studied the toxicity of P-Dichlorobenzene(P-DCB). The results showed that the toxicity of P-DCB was low. It has no accumulation (K > 5), no eye irritation and no skin sensitization effects. But light skin irritation effect was observed. In sub-acute inhalate exposure, damage on the function of liver and kidney was not observed. Genotoxic tests showed that tests in s. typhimurium strains TA98 and TA100 did not show any mutagenic potential. In Chinese hamster lung fibroblast CHL cell test system, it did not cause chromosomal aberrations. PMID- 11939022 TI - [Study on induced mutagenesis interaction of the high temperature and/or cigarette smoke]. AB - The study aim is to research the genotoxicity and interaction of the High temperature and/or cigarette smoke. The micronucleus of human lymphocytes and bone marrow in mice were observed following heat stress and/or watersoluble cigarette smoke substances. The results indicated that the high temperature (40 degrees C, 41 degrees C and 42 degrees C for 5 min, respectively) significantly induced micronuclei in human lymphocytes in vitro (P < 0.01). A significant synergistic effect of the high temperature and cigarette was found. When high temperature combining with high concentration cigarette smoke (1.0-2.0 cigarette/20 g/d, for 10 d) was used, a significant induction and interaction (P < 0.01) on micronucleus of the bone marrow were also observed. The results indicated that high temperature substanance aberrations would be enhanced when the high temperature combines with cigarette smoke. PMID- 11939023 TI - [Study on the changes of the levels of SOD and MDA in lung and MN in bone marrow exposed to cooking fume in rats]. AB - In order to investigate the mechanism of the activities of superoxide dismutase (SOD) and the concentration of malondialdehyde (MDA) in lung and rate of micro nucleus (MN) in bone marrow by which rape oil fume condensate(RC) exert their effect in rat lung, pouring into traches SD rat was performed with RC. Results showed that the SOD activities in RC and B(a)P groups were significantly higher than those in DMSO and control groups(P < 0.05). The concentrations of MDA were significantly lower than that in DMSO and control groups (P < 0.05). In experimental groups SOD/MDA decreased. MN rate significantly increased. It is concluded that lung tissue may be a target of RC. PMID- 11939025 TI - [Determination of methanol in hair spray by GC method]. AB - This report improved the chromatographic conditions of present standard method for testing methanol in hair spray GB7917.4-87. To avoid error detection on peak overlapping between methanol and dimethyl ether of original standard method, this method used column GDX-102 packed with 25% polyglycol 1540. The recoveries were in the range of 90.4%-107.0%, and the relative standard deviation was between 1.8% and 5.2%. PMID- 11939024 TI - [Effects of smoking on interleukin-2 and its receptor system in human peripheral blood]. AB - 42 healthy male smokers and 42 healthy male nonsmokers matched in age and life style were strictly chosen into the study from Beijing urban and rural areas. Peripheral blood mononuclear cells(PBMCs) were separated and then stimulated by PHA under 37 degrees C, 5% CO2 condition. Immunochemical methods were applied in testing the synthesis of interleukin 2(IL-2) and the expression of its membrane receptor (mIL-2R) and the level of soluble receptor (sIL-2R) in serum. The results showed that when compared with nonsmokers, stimulated PBMC from smokers had significant higher percentage of mIL-2R positive cells (P < 0.05) in vitro. Levels of sIL-2R in serum of smokers were also significantly higher than those of nonsmokers (P < 0.05). The percentage of IL-2 secrete cells of smokers had higher tendency (P = 0.08). These data indicated that the immune function of smokers and nonsmokers are different concerning the IL-2 and its receptor system. PMID- 11939026 TI - [Emission model of volatile organic compounds from materials used indoors]. AB - Various materials, such as wall-paper, floor-wax, paint, multicolor wall-coat, air freshener and mothball were experimented in a simulated test chamber under constant selected temperature, humidity and air exchange rate. The relation between the total VOCs concentration and time was regressed by four emission models and the surface emission rate was calculated. The regressed results indicated the similarity among four emission models for the liquid materials with volatile-solvent such as paint and multicolor wall-coat. But for low volatile solid materials, such as wall-paper, floor-wax, mothball, the sink model and the empirical model were better than the dilution model and vapor pressure model. Only for air freshener, it was improper to the total VOCs concentration as a parameter. PMID- 11939027 TI - [Effects of glutamic acid on heat tolerance and 70 kD proteins of Drosophila melanogaster]. AB - From supplement of dietary glutamic acid, the heat tolerance ability of Drosophila and the change of Hsp70(heat shock proteins) were studied. The results showed that the supplement of dietary glutamic acid could increase the survival time and the contents Hsp70 of Drosophila, in the 37 degrees C heat environment; but could not get the Hsp70 rise up when Drosophila were in 25 degrees C. Based on the findings, we could make the conclusion that supplement dietary glutamic acid could benefit the heat tolerance ability of Drosophila, and it might be related to the increase of Hsp70. PMID- 11939029 TI - [Development of an in vitro model of culture using mouse fetal long bone]. AB - An in vitro model of culture using mouse fetal long bone was firstly established using self-made turning-equipment. The bone formation, mineralization and resorption were observed simultaneously in cultured bone. It provided a useful model for investigating the relationship between bone, development and the effect of nutrients, toxicants and other factors. PMID- 11939028 TI - [Kinetic effect of testosterone or estradiol on iodine absorption in castrating rat intestine]. AB - OBJECTIVE: To observe the effect of testosterone or estradiol on iodine absorption in rat intestine. METHOD: 50 male adult Wistar rats were divided into 5 groups randomly. 50 females were divided into another 5 groups. Among them, 4 groups were bilaterally testectomized or ovariectomized, 1 group was sham operated. 7 days after operation, the castrated rats received testosterone (male rats) or estradiol (female rats) at different dosages by intramuscular injection for three days. Then the kinetics of iodine absorption in jejunum and ileum were observed by perfusion in situ. When finished, serum were obtained for detecting TSH, T4 and testosterone or estradiol. RESULTS: In castrated male rats, the value of K12 reduced, K21 increased, K02 reduced, and SP1/2 (the half time of the slow phase) prolonged, implying that the ability of iodine absorption reduced. It reflected that testosterone could promote iodine absorption in intestine in physiological condition. In castrated female rats, the situation was different from that in male rats, the value of K12 increased, K21 reduced, K02 increased, and SP1/2 shortened in jejunum, implying that the ability of iodine absorption increased. It reflected that estradiol could inhibit iodine absorption in intestine in physiological condition. The levels of serum TSH and T4 were not changed significantly in this experiment. CONCLUSION: In physiological condition, testosterone can promote iodine absorption, while estradiol has the inhibiting effect. The results indicate that gonadol hormone maybe one factor which can influence iodine absorption in intestine. It may explain the phenomenon that the incidence of goiter is different between males and females partly. PMID- 11939030 TI - [Preventive and therapeutic effects of calcium lactate on osteoporosis in aged ovariectomized rats]. AB - Twelve-month-old rats were ovariectomized (OVX), remained untreated for one month. Calcium lactate was given daily by gavage at the dose of 125, 250 and 750 mg/kg(Ca125, Ca250, Ca750) for 3 months. The results showed that compared with OVX rats, Ca250 could markedly increase the bone mineral density (BMD) at right femur. Both Ca250 and Ca750 could markedly enhance the ash weight, ratio of ash weight to dry weight of left femur and ratio of 24-hour urinary Ca/Cr. Ca125 could markedly decrease serum ALP concentration. Both Ca125 and Ca250 could decrease the ratio of 24-hour urinary Hop/Cr. All doses of calcium lactate groups could elevate serum Ca, and were no influence on the volume, wet and dry weight of the left femur and serum OC levels. In conclusion, the administration of calcium lactate to aged OVX rats prevents bone resorption, and its mechanism may be relating to the supplement of calcium properly. PMID- 11939032 TI - [Investigation on iodine contents in salt on market and in household]. AB - In order to supervise the iodine contents in salt on market and in household, a nationwide survey supported by ILSI was conducted. 91 salt samples on market and 531 samples in household were respectively collected from 35 manufactures in 11 selected provinces and from 165 districts in 25 provinces. The analytical method of determining the iodine in salt was based on Ce-As-I catalytic reaction. It was found that the iodine contents of the examined samples varied considerably and ranged from 3.0 to 100.3 mg/kg salt for market samples and 0.0 to 90.0 mg/kg for households samples. 71.4% and 73.4% samples reached the national standards respectively. 9.9% of market samples and 13.4% of the total 531 samples from households were lower than the national standard, while 18.7% and 13.2% of samples was higher. The percentage of coarse salt with low/no iodine content was 57.1%. PMID- 11939031 TI - [Effect of selenium status on the morbidity of interstitial pneumonia in rats infected with mycoplasma]. AB - To investigate the effect of selenium(Se) status on morbidity and process of interstitial pneumonia, we used the Wistar rats as the animal model infected with Mycoplasma Pneumonia (MP). The rats were maintained on the based diets with different doses of Se, infected with MP and exposed to sodium selenite(1.5-2.0 ml/day, containing Se 2 micrograms/ml) or 0.9% NaCl (1.5-2.0 ml/d) by gavage for different time. The experimental rats were divided into five groups, group A, feeding with normal control diet and without infected MP, group B with adequate Se diet and infected MP, group C with Se deficiency diet infected MP and added Se at the same time, group D with Se deficiency diet and added Se after suffered with MP, and group E with Se deficiency diet and infected MP. The results showed that the MP morbidity of the rats fed with high Se diet was significantly lower than that of the rats fed with low Se diet. The pathological change was significant in the low Se group and relatively mild in the high Se group. After infected with MP, the supplementation of sodium selenite to the low Se rats might decrease the MP morbidity and shorten the disease course compared with the group without supplementing Se. The incidence of myocarditis in the infected group with normal Se level diet or in the infected group with Se supplement was lower than that in the group without Se supplementation. The concentrations of Se in plasma and glutathione peroxidase in plasma and white blood cells decreased to a certain level after infection. The present study indicated a positive influence of Se supplement on the morbidity, disease course, and state of interstitial pneumonia in rats. PMID- 11939033 TI - [Effects of cooking methods on iodine content in iodized salt]. AB - Effects of cooking methods and variety of foods on the retention of iodine content in food with iodized salt were studied. Vegetables from market and usual cooking methods were selected, including procedures of various cooking methods. The samples were fixed by potassium carbonate, ashed with zinc sulfate at 550 degrees C, then determined by colorimetric method with Ce-As-I catalytical reaction. The different cooking methods had different effects on the retention of iodine, in general, the retention of iodine by stewing of steaming was higher than by stir-frying. The effect of various vegetables on the retention of iodine was also different. The retention of iodine after stir-frying was 84.2%, 56.9%, 44.5%, 36.6% for fruit-bearing vegetables, leafy vegetables, roots and beans. The retention of iodine after stewing was 66.1%, 53.4%, 47.0%, and 43.2% for fruit bearing vegetables, roots, beans and meat. The stability of organic iodine in food is higher than that of inorganic iodine. PMID- 11939034 TI - [Effect of allicin on ethanol-induced hepatotoxicity in mice]. AB - To study the effect of allicin, an effective component of garlic, on ethanol induced hepatotoxicity in mice. The results showed that allicin (10 mg/kg ig, qd x 10) could reverse the higher activities of serum ALT and glutathione s transferase (GST) in ethanol-treated mice. Furthermore, allicin could significantly enhance the content of hepatic reduced glutathione (GSH), and the activities of hepatic glutathione peroxidase (GSH-Px), glutathione reductase (GSH Re) and GST in ethanol-induced hepatotoxicity mice. There were no remarkable changes in the hepatic catalase (Cat) and superoxide dismutase(SOD) activities. These results suggested that allicin have the effective hepato-protection on ethanol-induced hepatotoxicity, which is related to its selective effect on the glutathione-related enzyme system. PMID- 11939035 TI - [Determination of toxic peptides in Amanita virosa and Amanita verna]. AB - The toxic peptides in Amanita virosa and Amanita verna from various region were analyzed by a reversed-phase high-performance liquid chromatography. It is showed that there were great differences in the concentrations of toxic peptides between species and strains of A. virosa and A. verna. PMID- 11939037 TI - [Study on the analytical method for organochlorine pesticide residue using gas chromatography-mass spectrometry]. AB - A gas chromatographic-electron ionization--mass spectrometric method (GC-EIMS) was developed for determining organochlorine pesticide residues. Ten organochlorine pesticides including alpha, beta, gamma and delta isomers of hexachlorocyclohexane(HCH), pp'-DDE, pp'-DDT,op'-DDT,pp' DDT,pentachloronitrobenzene (Quintozene, PCNB) and hexachlorobenzene(HCB) were determined respectively. Targeted pesticide analyses were separated on DB-5 capillary column. The optimum chromatographic conditions and MS parameter were determined. In test condition, each constituent in the standard mixture was separated efficiently. The retention time of each compound and its characteristic ion were determined in fullscan mode from 50 to 450 amu. The values of their mass spectrum on the total ion current(TIC) with fullscan mode agree with those of NIST and pesticide libraries. The characteristic ions were selected to quantify at m/z 181 for the isomers of HCH, m/z 284 for HCB, m/z 295 for PCNB, m/z 246 for pp'-DDE, m/z 235 for pp'-DDD, op'-DDT and pp'-DDT. The accuracy and reliability were proved respectively by linear test, precise test and recovery test. The linear ranges of PCNB and pp'-DDT are 0.0025-0.50 microgram/ml. The ranges of the isomers of HCH, pp'-DDE, pp'-DDD and op'-DDT are 0.0020-0.40 microgram/ml. The limits of detection are 0.0025 ng for PCNB and pp'-DDE, but 0.0020 ng for the isomers of HCH, HCB, pp'-DDE, pp'-DDD and op'-DDT. PMID- 11939036 TI - [Effect of Ciwujia (Radix Acanthopanacis senticosus) preparation on exercise performance under constant endurance load for elderly]. AB - The effect of Ciwujia (Radix Acanthopanacis senticosus) preparation on human exercise performance was investigated by using 13 healthy volunteers aged 50-57. Under constant endurance load with 450 kg.m/min (75 W), the respiratory quotient after taking Ciwujia preparation was reduced to 0.88 from 0.96, which implied that the utilization of lipid increased with 27.2% as energy fuel during exercise. Meanwhile, the heart rate was reduced with 8.7%, and O2 uptake per heart beat increased with 16.18%. All results indicated that the Ciwujia preparation could increase O2 uptake, spare the glycogen in muscle and improve the exercise endurance and work performance in human subjects. PMID- 11939038 TI - [Effect of zinc on the cellular immunity and NK cell activity in acute heat exposed rats]. AB - To study the effects of zinc on the cellular immunity and NK cell activity in acute heat-exposed rats. Rats were fed with dietary containing high, moderate and low zinc for 15 days. The concentrations of zinc in high, moderate and low zinc dietary are 92.20 mg/kg, 45.61 mg/kg and 21.70 mg/kg respectively. After exposed to acute heat (41.5 degrees C) for 5 min, 30 min and 60 min, the rats were killed and the cellular immunity and NK cell activity were measured. The proliferation of lymphocyte and the NK cell activity were higher in high zinc group than in moderate and low zinc groups no matter the rats were exposed to heat or not. Zinc can improve the cellular immunity and NK cell activity in acute heat exposed rats. PMID- 11939039 TI - [Studies on the Ras p21 levels in workers exposed to coal tar pitch and coke oven volatiles]. AB - To study the effects of serum p21 levels on the health of coal tar pitch(CTP) and coke oven volatiles (COV) workers recognized as the high risk population of pulmonary carcinoma, serum p21 levels among 24 cases of CTP workers, 25 cases of COV workers and 12 cases of normal control were detected with the method of Western Blotting. The results showed that the mean serum p21 level in exposed group was significantly higher than that in the normal control (366.8 vs 1270.7, P < 0.001). In the COV workers, serum p21 level in coke furnace workers was remarkably higher than that in the workers with other types of work. There were no differences for p21 among workers with different work years and between workers with or without smoking history. It indicates that CTP and COV can cause the p21 protein level increase in the occupational population. PMID- 11939040 TI - [Study on the joint effects of acrylonitrile and alcohol in rats]. AB - In order to learn the joint effects of acrylonitrile and alcohol, acrylonitrile(40 micrograms/g), alcohol (1000 micrograms/g) and the mixed liquid of the both (40, 1000 micrograms/g) were given to rats by gavage for 8 weeks. The results revealed the joint effects of acrylonitrile and alcohol. Hypofunction and structural lesion of hepatocyte were found in the rats. The numbers of white blood cells decreased remarkably in these two groups. PMID- 11939041 TI - [Effects of cadmium on 3H-leucine incorporation and cell growth of cultured myocardiocytes]. AB - The effects of cadmium on 3H-leucine incorporation and cell growth of cultured cardiac myocytes were investigated by measuring the incorporation quantity of radioisotope labelled amino-acid and the cell volume on cultured myocardiocytes in neonatal Wistar rats. The results showed that 3H-leucine incorporation was decreased significantly in 1 x 10(-4) mmol/L CdCl2 for 24 h, and was gradually reduced with prolongation. The cell volume was decreased after exposed to Cd for 48 h. The volume was from (2053 +/- 189) microns 3 to (1847 +/- 176) microns 3. There was no significant difference in cell number between the treated group and the control group. These data indicated that cadmium can inhibit the synthesis of protein and the growth on cultured myocardiocytes. PMID- 11939042 TI - [Reference value of urinary arsenic for nonoccupational exposed subjects in Henan]. AB - Urinary arsenic levels of 2065 nonoccupational exposed subjects in Henan were determined by spectrophotometric analysis. The results showed: (1) The difference was not significant among Zhengzhou, Luoyang, Anyang, Xinyang and Shangqiu (0.01 < P < 0.025). (2) The difference was not significant among different ages(0.01 < P < 0.025) and between sexes(P > 0.05). (3) The distribution was positive skewness. The median of urinary arsenic was 17.8 micrograms/L. The arithmetic mean was 20.7 micrograms/L. The standard deviation was 15.2 micrograms/L. The geometric mean was 17.3 micrograms/L. (4) It is suggested that the reference upper limit value of urinary arsenic for nonoccupational exposed subjects in Henan in 56 micrograms/L. PMID- 11939044 TI - [Study on the genotoxicity of methyl tertiary-butyl ether]. AB - Methyl tertiary-butyl ether(MTBE), a new gasoline additive, is added to gasoline as an octane enhancer and used to reduce automotive emissions. Using Ames test, unscheduled DNA synthesis (UDS) assay and cell micronucleus test, the authors evaluated the genotoxicity of domestic produced MTBE. MTBE did not show any mutagenic potential in Ames assay(TA98, TA100 strains) with or without rat liver derived metabolic activation system (S-9). Rat primary hepatocyte UDS assay showed that MTBE could damage DNA. In NIH3T3 cell micronucleus test, MTBE showed negative results. It could be concluded that MTBE has some genotoxicity at DNA level. PMID- 11939043 TI - [Studies on molecular teratogenic mechanism of methylmercury in early developing rat embryos]. AB - The present study further revealed the teratogenic mechanism of methylmercury chloride (MMC) during rat neurulation by means of nonradioactive in situ hybridization (ISH), semiquantitative analysis of immunohistochemical staining and in vivo teratogenic test. The main aim was to teat the hypothesis that iNOS, HSP70, NT, TGF-beta and Bcl-2 genes contribute to MMC-induced day 9.5 embryonic damages. Results showed there were no obvious poisoning signs and death of pregnant female rats injected intraperitoncally with 0, 0.2, 0.4, 0.8, 1.6 and 3.2 mg/kg MMC. While the doses increased, the total morphological scores decreased gradually, and the rates of embryo deformity and development delay increased step by step to 34% and 76% respectively. The levels of iNOS mRNA and protein and HSP70 mRNA increased, and NT mRNA with its protein became down, all these changes were concentration dependent. In addition, MMC could inhibit the level of TGF-beta mRNA, but no obvious influence on the levels of Bcl-2 mRNA and Bcl-2 protein. On the basis of parallel findings from embryos, genes and proteins, abnormal expression of genes in transcriptional level might be related to MMC-induced teratogenic insult. PMID- 11939045 TI - [Joint effects of arsenic and lead on lipid peroxidation of human and rat erythrocytes]. AB - The authors observed the joint effects of arsenic and lead on lipid peroxidation(LPO) in vitro and in vivo. In vitro, the authors used the extracted erythrocyte membrane of health adult men and divided them into three exposed groups. The first group was exposed to As2O3 (0, 0.4, 2.0 and 10.0 mmol/L). The second group was exposed to PbAc2 (0, 0.08, 0.4 and 2.0 mmol/L). The third group was exposed to As(10 mmol/L) + Pb (0.08 mmol/L). The results show that there is interaction between As and Pb on LPO of erythrocyte membrane. In vivo, the authors measured LPO levels of rat's plasma, blood glutathione(GSH) and superoxide dismutase(SOD) activity. By using 2 x 2 factorial design, Wistar rats were gavaged with As2O3 2.4 mg/(kg.d) and PbAc2 30 mg/(kg.d) as well as both arsenic and lead in water for 10 days. The results show that in the As-Pb treated rats, the levels of LPO did not change significantly, the GSH levels and SOD activity decreased significantly. The results of factorial analysis revealed a synergic effect of As-Pb on SOD activity. PMID- 11939046 TI - [Comparison of plasma fibrinogen level in members of Uygur family of longevity and non-longevity in Xinjiang]. AB - OBJECTIVE: The associations of plasma fibrinogen level with age in members from longevity families and non-longevity families were compared and the influences of relationship in family members to plasma fibrinogen level were analysed to study the relationship between plasma fibrinogen level and life-span run in family. MEANS: The authors measured fibrinogen in 156 Uygur subjects from 7 longevity families and 9 non-longevity families. RESULTS: Fibrinogen level was relatively stable among different age groups in longevity families. It was an association positively with age in non-longevity. Fibrinogen level of children was closely correlative with parent's. CONCLUSIONS: Genetic factor probably contributes to plasma fibrinogen level. Plasma fibrinogen level correlates with life-span run in family. PMID- 11939047 TI - [Epidemiological study of airway responsiveness on the patients with asthma]. AB - The analysis of date on 641 nuclear families with asthma declared that the sensity and specify of the airway responsiveness with Methecholone(MTCH) Challenge Test for the clinical diagnosis of asthma is 72 percent and 70 percent. The population with airway hyperresponsiveness was 6.01 times higher risk to asthma than one of negative airway responsiveness. The OR of male and female for asthma was 4.97, 5.06, respectively, and was statistically significant (P < 0.01). Moreover, the risk of asthma was been increasing with MTCH concentration was increased (i.e. degree of airway responsiveness decreased) step by step, as well showed the lineage trend between the asthma and airway responsiveness significantly (P < 0.01). The Measurement of airway responsiveness play an important role in clinic and prevent medicine. PMID- 11939048 TI - [Effects of different zinc level diets and heat exposure on the contents of Zn, Cu and Fe in brains of rats]. AB - Forty-eight SD male rats were randomly divided into group A, B and C. Rats in each group were also divided into 2 parts. One part was used for heat exposure and the other was used for room temperature control. On the basis of AIN-93 m diet composition, group A, B and C zinc diets were prepared by adding or reducing zinc carbonate. Zinc contents of group A, B and C zinc diet per kilogram were 92.2 mg, 45.61 mg and 21.70 mg respectively. 14 days later the rats were exposed to high temperature (Tdb 40 degrees C, Twb 30.8 degrees C) for 3 hours. The Contents of Zn, Cu and Fe in hippocampus, cerebral cortex and cerebellum of rats were determined by atomic absorption spectrophotometer. The results revealed that 1. hyperthermal exposure could increase Cu content in rat hippocampus significantly, 2. after heat exposure, Cu content in hippocampus was higher than in cerebral cortex and cerebellum. Cu contents in cerebellum of room temperature control rat were higher than cerebral cortex and hippocampus, 3. intakes of different zinc level diets chiefly influenced content of Cu rat cerebellum and slightly affected iron content in it, 4. high, middle level zinc diet intake could reduce Cu content in heat-stressed rat cerebellum. PMID- 11939049 TI - [Induction of NAD(P)H: quinone reductase by anticarcinogenic ingredients of tea]. AB - By assaying the activity of NAD(P)H: quinone reductase (QR) in Hep G2 cells exposed to inducing agents, a variety of ingredients in tea, we compared their abilities on inducing QR and preventing cancer. The results showed that tea polyphenols, tea pigments and mixed tea were all able to induce the activity of QR significantly. The single-component ingredients of tea polyphenols and tea pigments, including thearubigens, EGCG and ECG, also enhanced the activity of QR. But EGC, EC, theaflavins, tea polysaccharide and tea caffeine, showed no apparent induction of QR. We found that among those tea ingredients studied, the multi component ingredients were more effective than the single-component ones. So we thought that the abilities of antioxidation and cancer prevention of tea depended on the combined effects of several kinds of active ingredients, which mainly include tea polyphenols and tea pigments. PMID- 11939051 TI - [The food habit and its affecting factors of preschool children in Guangzhou]. AB - A survey of nutritional knowledge-attitude-practice was conducted among 1300 preschool children, their parents, 203 teachers and nurses in Kindergarten in 1997. Results showed that the food habit of preschool children was unsuitable. Most preschool children rejected eating animal liver, peanut product and animal blood, but often ate snacks. From statistic analysis among affecting factors, the mother's and teachers' nutritional knowledge and the food habit of parents had significant effect on food habit of preschool children. The family income did not significantly affect food habit of preschool children. Moreover, there was little nutrition education involved in the preschool children courses in 4 kindergartens. The result suggested that the nutrition education plan should be done. PMID- 11939052 TI - [Nutritional status of the residents in past 10 years in Ningxia Hui autonomous region]. PMID- 11939050 TI - [The impact of maternal education on the dietary and nutritional status of preschool children--a case study in 8 provinces of China]. AB - This analysis is based on the data from China Health and Nutrition Survey in collaboration with University of North Carolina at Chapel Hill undertaken in 8 provinces of China from 1993 to 1996. Totally, 1098 mother-child pairs under age 6 were investigated. Descriptive analysis and stratified analysis are applied to investigate the impact of maternal education on the dietary and nutritional status of children. Both the crude analysis and after controlling for confounding variables, maternal education in the population remains statistically relevant to children's dietary and nutritional status. The proportion of low body weight and stunting children is the highest in the group with maternal education level being below preliminary school, the lowest in the group with education level being above high middle school. Moreover, in these three groups, the higher the level of maternal education, the greater the percentage of nutrients intake of China RDA, especially those from animal food. These results fully illustrate the importance of maternal education for the prevention of malnutrition in children. PMID- 11939053 TI - [Solid-phase microextraction of dichlorvos from white spirits]. AB - DDVP was isolated from white spirits using Solid-Phase Microextraction (SPME) technique, and determined by wide-bore capillary gas chromatography (GC) coupling a flame ionization detector. This method had an excellent linearity among 0.5-12 mg/L. Correlation coefficient and minimum detectable concentration were 0.9983 and 0.05 mg/L respectively. This method enables the extraction, the enrichment and the injection to integrate into a single step, and it was free-solvent in whole process. SPME is an advanced technique for the extraction and analysis of the samples. PMID- 11939054 TI - [Effects of taurine and extraction of cristata L on serum Zn, Cu and Ca in rats]. AB - Forty rats fed with basic diets were randomly divided into 4 groups. NG group were fed with basic diets. The other three groups were fed with high fat diets. The rats in TA group and EC group were given TA 100 mg/kg or EC 100 mg/kg each day respectively in addition to high fat diet for 8 weeks. The results showed that taurine and extraction of cristata L not only increased the level of red cell SOD and the content of serum Zn (P < 0.05), but also decreased the contents of TC, MDA in the wall of artery and decreased the level of serum LDH significantly (P < 0.05 or P < 0.01). TA and EC increased significantly the content of serum Cu and decreased the ratio of serum Cu to Zn in the high fat diet rats (P < 0.01), and decreased the contents of serum Ca also. The results indicate that TA and EC may play some role in lipid metabolism and inhibit atherosclerosis by regulating the levels of Zn, Cu and Ca in rats. PMID- 11939055 TI - [Effect of selenium on children suffered from Mycoplasma pneumonia]. AB - In order to evaluate the therapeutic effects of Selenium (Se) on Mycoplasma pneumonia, a clinical study was conducted in hospitalized patients using a double blind controlled method. The patients were randomly divided into two groups, the control patients received the routine treatment, and the supplemented group was given 1 mg sodium selenite one time only in addition to the routine treatment. The present study showed that concomitant of Se and routine treatment was superior to routine treatment alone in terms of improvement of clinical signs, time needed for relieving symptoms and signs (P < 0.05). The levels of Se and glutathione peroxidase in plasma and white blood cell were increased in Se supplemented group (P < 0.05). Higher proportion of patients in the treated group showed normalization of T cell subset parameters as compared with the control group. It can be concluded that supplementation of Se would be conducive to the improvement of the nutritional status and the increase of the ability of resisting-infection. PMID- 11939056 TI - [Study on fugu medicated therapeutic prescription]. AB - The Fugu medicated therapeutic prescription (FGMTP) composed of the medicinal herbs with food usage was investigated on the nutrition components and its anti tumor activity. The results showed that the ratios of thermogenesis of three calorifacient nutrients were protein (12.34%), fat(5.62%) and carbohydrates (82.0%), respectively. The IC50 values of the water extract from this prescription against the 7901 or K562 cell line colony formations were 3.59 g/L and 25.74 mg/L, and the maximal inhibitory rates against both cell line colony formations were 53.2% and 58.9%; respectively. The maximal inhibitory rates against HepA and S180 transplantation tumors were 35.3% and 35.0%, respectively. It is concluded that the FGMTP possessed obvious antitumor effects. PMID- 11939057 TI - [Preparation of morchella liquid by submersed fermentation and and analysis of its nutrient contents]. AB - Natural morchella esculenta fruits were collected and its spores were isolated to get pure mycelium. Morchella liquid was prepared by submersed fermentation technology. The quantitative analysis showed that the liquid is relatively high in nutrient contents and several kinds of other essential components. The nutrient contents of the liquid are as follows (g/L): carbohydrates 63.1, reduced saccharide 25.6, protein 8.3, dry material 104.7, minerals 1.5. There are also several high active micromaterials as follows: morchella lentinan 1.45 g/L, 10 kinds of essential and semiessential amino acid, 12 kinds of mineral (Fe, Zn Se, Cr, etc.). The amount of soluble proteins are high in the liquid(one wider zone and four narrow zones on electrophoresis with molecular weights of 31.0 kd, 97.4 kd, 66.2 kd and 52.9 kd respectively). The morchella liquid can be used as a new kind of base material for health foods and a flavoring agent. PMID- 11939058 TI - [Identify influential points]. AB - Influential points are the points with excessive influence on result and detected by influential function theta-theta(i). theta-theta(i) is a multidimensional vector. For convenience, people always consider some quantitative function g (theta-theta(i)) to measure the vector theta-theta(i), but how to define g is a difficult problem. This paper proposes a new quantitative diagnostic statistics for identifying influential points for all parametrical statistical analysis. The method is more appropriate than cook's method for regression analysis. In this study an example is used to illustrate the procedure in COX proportional hazard model. PMID- 11939059 TI - [Deiodinase]. AB - The members of deiodinase family including type I, type II and type III deiodinase are seleno-proteins. They play an important role in the regulation of thyroid hormone metabolism. They are different in substrate specificity and sensitivity to inhibitor and kinetics mechanism. Their cDNAs exhibit limited regions of homology, but they have common conserved sequence, TGA codon of selenocysteine and some structures and sequences which are essential for selenocysteine insertion. PMID- 11939060 TI - [The anti-ischemic effect of O, O-acetyldaurisoline]. AB - O, O-acetyldaurisoline(Adau) is a derivative of daurisoline (Dau). In cultured PC12 cells, Adau was found to inhibit K+, Bay K8644 and norepinephrine induced intracellular free calcium concentration increase. Adau was also shown to protect PC12 cells from hypoxia-reoxygenation injury with EC50 of 64.3 +/- 12.5 mumol.L-1 by MTT assay. Adau (2.5, 5 and 10 mg.kg-1, i.v.) administrated 30 min before ischemia in bilateral carotid artery occlusion and four-vessel occlusion rats, attenuated the increase of lipid peroxide content and the decrease of SOD activity. These results show that Adau have significant protective effects against ischemic injury. PMID- 11939061 TI - [Protective effects of dl-3-n-butylphthalide on changes of regional cerebral blood flow and blood-brain barrier after subarachnoid hemorrhage]. AB - Effects of dl-3-n-butylphthalide(dl-NBP) on changes of regional cerebal blood flow (rCBF) and blood-brain barrier of rats subjected to subarachnoid hemorrhage (SAH) were investigated. Regional CBF was determined by hydrogen clearance method at 15 min before, 15 and 30 min thereafter once every 30 min up to 180 min after the beginning of SAH. The results showed that SAH produced a significant decrease in rCBF in caudate nucleus and blood-brain barrier damage which displayed increased extravasation of Evans blue to brain tissue. Both dl-NBP (5-20 mg.kg-1 given intraperitoneally 5 min after the onset of SAH) and nimodipine (0.25 mg.kg 1) were shown to increase rCBF in caudate nucleus through 3 h of SAH. Moreover, dl-NBP 10 mg.kg-1 and nimodipine 0.25 mg.kg-1, given 5 min and 3 h, respectively, after SAH, significantly reduced the brain extravasation of Evans blue 6 h after SAH, indicating that both dl-NBP and nimodipine has protective effects on blood brain barrier. These results suggest that dl-NBP has therapeutic effects on SAH. PMID- 11939063 TI - [Difference between the effects of (-) clausenamide and (+) clausenamide on the synaptic transmission in the dentate gyrus of anesthetized rats]. AB - With the extracellular recording technique, the basal synaptic responses (population spike, PS) evoked by low frequency test stimulation in the dentate gyrus of anesthetized rats was recorded and the effects of (-) and (+) clausenamide(icv) on the PS and long-term potentiation (LTP) induced by tetanus (50 pulses at 200 Hz) were observed. The results showed that: (1) at a lower dose (1 nmol), (+) clausenamide showed no effect on either the basal PS or LTP, (-) clausenamide did not affect the basal PS but enhanced the magnitude of LTP. (2) At a higher dose (4 nmol), (-) clausenamide not only potentiated the basal PS but also dose-dependently augmented the magnitude of LTP. Meanwhile, (+) clausenamide did not affect the basal PS but attenuated the magnitude of LTP. These results suggest that the effects of clausenamide on the synaptic transmission in the dentate gyrus depend on its chirality. The potentiating effects of (-) clausenamide on synaptic transmission in hippocampus strongly support our previous behavioral and neurobiochemical studies on the nootropic action of (-) clausenamide. PMID- 11939065 TI - [Sulfation of polysaccharides isolated from Indocalamus tesselatus and their anticytopathic effect on human immunodeficiency virus type I]. AB - For preparation of sulfated polysaccharides isolated from bamboo leaves (Indocalamus tesselatus) with chlorosulfonic acid-pyridine, the experimental conditions as the molar ratio of reactants and temperature and time of reaction were discussed. A homogeneous sulfation method with good yield of more than 80% was obtained and the sufur content was found to be 21.9%. Conformation change in sulfated polysaccharide with changes of intrinsic viscosity, polarimetery and CD spectra were found to be related to the negative charge. The polysaccharide sulfates were shown to have better activity to inhibit cytopathic effect of human immunodeficiency virus(HIV) infection than native polysaccharides. The effective concentration was 10 micrograms.ml-1 while the cytotoxic concentration was 2.5 mg.ml-1. PMID- 11939066 TI - [Studies on synthesis and bioactivity of 2-alkenyl-4(1H)-quinolone]. AB - Three 2-alkenyl-4(1H)-quinolone compounds(I-III) were synthesized by two methods. III is a new compound from Evodia rutaecarpa and was synthesized through the reaction of the aldehyde intermediate(5) with Wittig reagent. The other two compounds were synthesized for the first time. These compounds showed vasodilating and antibacteria effects in pharmacological tests. PMID- 11939064 TI - [Effects of (-), (+)clausenamide on anisodine-induced acetylcholine decrease and associated memory deficits in the mouse brain]. AB - The effect of a new cognition enhancer clausenamide on regional acetylcholine (ACh) levels and anisodine-induced ACh decrease were examined in mice of memory deficits. In the mean time, the effect of (-)clausenamide on ACh was compared with that of (+)clausenamide. Animal amnesia model was induced by i.p. anisodine, brain acetylcholine content was measured by high performance liquid chromatography with electrochemical detection. It was found that single administration of (-)clausenamide or (+)clausenamide(10, 20, 50 mg.kg-1, ig) showed no effect on the ACh level in the frontal cortex, hippocampus and striatum in normal condition. However, pretreatment with (-)clausenamide (10, 20, 50 mg.kg 1, ig) significantly ameliorated the reduction of ACh in these regions induced by anisodine (10 mg.kg-1, i.p.) in a dose-dependent manner. In the meantime, (-) clausenamide ameliorate anisodine-induced memory deficits in step-through test in mice. In contrast, (+)clausenamide showed no effect on these sides. The results indicate that there is significant difference between the actions of ( )clausenamide and (+)clausenamide; The protective action of (-)clausenamide against anisodine-induced amnesia is due to its ability to reverse ACh reduction. PMID- 11939067 TI - [Study on the polyoxygenated cyclohexenes from Uvaria boniana]. AB - Seven new polyoxygenated cyclohexenes, named uvaribonol A-G (1-7) have been isolated from the ethanol extract of the stems of Uvaria boniana Finet. (Annonaceae), and their structures, including the absolute configuration, were determined by spectral and chemical methods. In vitro cytotoxicity test against several human tumor cell lines indicated that all of the new natural compounds are inactive, but some of the derivatives showed obvious activities. Compound 2a is the most active, exhibiting significant cytotoxicities against KB and Bel7402 cells with IC50 < 1 microgram.ml-1, and against HCT-8 cell with IC50 < 0.1 microgram.ml-1. PMID- 11939069 TI - [Determination of diflunisal in plasma by RP-HPLC after solid-liquid extraction]. AB - A RP-HPLC method was developed with solid-liquid extraction technique. Plasma sample was extracted on a macroreticular resin cartridge with methanol--glacial acetic acid (99:1) as elution solvent. After extraction, the assay was carried out on a Spherisorb C18 column with p-phenylphenol as internal standard. The mobile phase is a mixture of methanol--water--glacial acetic acid (66:30:4). UV detection was performed at 250 nm. The flow rate was 1.0 ml.min-1. A good linearity was found at the concentration range from 0.5 to 100 micrograms.ml-1, with the lowest detection limit 0.02 microgram.ml-1 (S/N = 2). The extraction and method recoveries were 91.65% and 97.25% respectively, while the RSD for the within-day and between-day precision were all less than 10%. The above method was applied to determine the plasma concentration of difunisal in three human volunteers after a single oral dosage of 300 mg. Two hours after administration, the plasma concentration of diflunisal reached maximum level. A two compartment method was used to study the pharmacokinetic parameters. The T1/2 alpha and T1/2 beta were 1.40 h and 17.85 h, respectively. PMID- 11939068 TI - [A new saikosaponin from Bupleurum chinense DC]. AB - Bupleurum chinense DC. is a well-known and very important traditional Chinese drug. It is often used to treat common cold with fever, alternating chill and fever, feeling of fullness and oppression in the chest. A new saikosaponin was isolated from Bupleurum chinense DC. and its structure was identified as 3-O [beta-D-glucopyranosyl (1-->3)-beta-D-fucopyranosyl]-3 beta, 16 alpha, 23, 28 tetrahydroxy-olean-11, 13 (18)-dien-30-oic acid 30-O-[xylityl(1-->1)]-beta-D glucopyranosyl-6-ester on the basis of chemical and spectral evidence. In addition, three known saikosaponins were isolated and characterized as saikosaponin 1, 2"-O-acetyl-saikosaponin b2 and 2"-O-acetyl-saikosaponin a. All the compounds were isolated from Bupleurum chinense DC. for the first time. PMID- 11939071 TI - [Metabolic transformation of (-)-clausenamide in rat liver microsomes]. AB - The metabolic transformation of (-)-clausenamide, isolated from the leaves of Clausena lansium (Lour.) Skeels, was studied in vitro with phenobarbital-induced rat liver microsomal incubate containing the NADPH-generating system. The constitution of the incubation system was optimized and a RP-HPLC-DAD method was developed for the on-line analysis of (-)-clausenamide and its metabolites. The major metabolites were isolated and purified by silica gel column chromatography, preparative TLC and HPLC and their structures were determined as 6-OH clausenamide (CM1) and 5-OH clausenamide (CM2) by 1HNMR and MS. PMID- 11939072 TI - [Determination of flavonoids and quality evaluation of Chinese traditional drug "puhuang"]. AB - This paper deals with the determination of flavonoids and quality evaluation of Chinese Typha. Four species, viz. Typha angustata, T. angustifolia, T. davidiana and T. latifolia and eight commercial samples of Chinese traditional drug "Puhuang" were collected. Five flavonoids in pollen, filaments, female flowers and leaves were determined by HPLC using mu-Bondapak C18 column, water- isopropanol--tetrahydrofuran = 83.5:14:2.5, as mokile phase and detected at 287 nm). The qualities of different Typha species and commercial "Puhuang" samples have been compared, and the criteria for their quality control were also discussed. PMID- 11939073 TI - [Molecular identification of the Chinese drug turtle shells]. AB - This paper reports a new method to identify the Chinese drug turtle shells using PCR product direct sequencing method. DNA was extracted from tissues of the Chinese three-keeled pond turtle Chinemys reevesii and 20 other species of turtles occurring in China and Southeast Asian countries. One hundred and ten base pairs of mitochondrial 12S rRNA gene fragment were amplified from the extract using PCR technique and obtained sequences. These sequences were used to construct 12S rRNA gene fragment sequence database for the 21 turtle species. Comparison of these sequences indicated that the sequence from the Chinese three keeled pond turtle is different from that of all the other 20 turtle species. The sequence divergence is 3.7-15.7%. DNA was extracted from 0.1-0.5 g of shell from 19 turtle shells provided by the Jiangsu Institute for Drug Control and 12S rRNA gene fragment was amplified and sequenced. Comparison of the sequences from the 19 turtle shells and 12S rRNA gene fragment sequence database indicated that only 3 samples are shells of the Chinese three-keeled pond turtle specified in the Phamacopoeia of the People's Republic of China and the others are substitutes. The technique used in the present paper was found to be effective and reliable for the identification of turtle shells. PMID- 11939074 TI - [Chemical reactivities and the core modifications of paclitaxel and its analogs]. PMID- 11939075 TI - Use of miniscrews for intermaxillary fixation of lingual-orthodontic surgical patients. PMID- 11939076 TI - A rapid molar intruder for 'non-compliance' treatment. PMID- 11939077 TI - Are your accounts receivable healthy and balanced? PMID- 11939079 TI - Promoting the auto-referral practice: your long-term success insurance. PMID- 11939078 TI - Molar control. Part 3. PMID- 11939080 TI - Using metal mesh for a temporary bonded prosthesis. PMID- 11939083 TI - From age-ing to sage-ing. PMID- 11939082 TI - Stigma in psychiatric nursing. AB - TOPIC: Stigmatizing attitudes in psychiatric nursing. PURPOSE: To develop a greater awareness of the existence of stigma associated with psychiatric nursing. SOURCES: Published literature. CONCLUSIONS: Professional self-understanding and knowledge are methods by which psychiatric nursing can elevate its professional image and value. PMID- 11939081 TI - Utilizing Ericksonian hypnosis in psychiatric-mental health nursing practice. AB - TOPIC: Ericksonian hypnosis conceptual framework. PURPOSE: To acquaint psychiatric-mental health nurses with hypnotic principles and how these can be integrated into their practice. SOURCES: Published literature and author's clinical experience. CONCLUSIONS: Ericksonian hypnosis offers an array of potential interventions for psychiatric-mental health nurses to integrate into their practices in a framework familiar to nurses: holism, honoring and respecting individuality, and capitalizing on an individual's strengths. PMID- 11939085 TI - Enhancing therapeutic presence. PMID- 11939084 TI - Psychotropic drug interactions with grapefruit juice. PMID- 11939086 TI - Nurses' grief reactions to a patient's suicide. AB - TOPIC: A patient's suicide may threaten the nurse's health and work performance until grief and mourning are transformed. PURPOSE: To examine the literature, bereavement theories, and recommendations for supporting nurses' bereavement. SOURCES: Bereavement literature on Medline, CINAHL, and PsychInfo from 1965-2001, and clinicians' and nurses' responses to a patient's death by suicide. CONCLUSIONS: Nurses need a support system to help them cope with grief after a patient's suicide. Having knowledge of bereavement and using therapeutic support can help prevent burnout or stress and can encourage constructive coping strategies that transform grief. Grieving is facilitated when nurses recognize their own mortality and take time to process their grief. PMID- 11939087 TI - Is there an association between SEN virus and liver disease? Reviewing the evidence. PMID- 11939088 TI - [Parasplenial arteriovenous malformations. Report of 15 surgically treated cases]. AB - INTRODUCTION: Yasargil called paraesplenial those AVMs located at the confluence of the hippocampus, the isthmus of the cingulate girus and the girus occipitotemporalis medialis. Large AVMs at this location are among the most difficult to delineate and to treat. OBJECTIVE: Analyze the clinical presentation, the findings in the imaging studies, the surgical management and the final outcome in 15 patients with paraesplenial AVMs treated with embolization (the last 4 cases), and microsurgical removal. RESULTS: Nine patients (60%) were female and 6 males of ages between 15 and 39 years (mean = 24 yrs). Eleven (75%) presented with hemorrhage, (intraventricular in most cases) and the remaining with epilepsy. The Spetzler-Martin grade was II in one Case, III in 5 cases, IV in 8 cases and V in one case. Preoperative embolization clearly improved surgical management. All the patients had complete resection of the lesion, 13 in a single stage and 2 in two stages. The final outcome was good but four patients developed defects of the visual field not seen preoperatively. CONCLUSIONS: The authors comment the peculiarities of paraesplenial AVMs which can be safely and completely removed with microsurgery and the aid of preoperative embolization. PMID- 11939089 TI - [Epidemiologic studies of cerebellopontine angle tumors surgically treated in Maracaibo, Venezuela, in 1985-1999]. AB - OBJECTIVE: To analyze the epidemiological, clinical and neuropathological data of cases of cerebellopontine angle (CPA) tumors. MATERIAL AND METHODS: The clinical records, neuroimaging and neuropathological studies of 50 patients with diagnosis of CPA tumor operated in different hospitals of Maracaibo, Venezuela, during the lapse January 1st, 1985-December 31, 1999 were reviewed. The variables age, gender, side of the lesion and neuropathological diagnosis were analyzed. RESULTS: A 2:1 female to male ratio was observed. Median age was 48 +/- 12.7 years. Acoustic neuromas (AN) represented 48% of the cases, whereas nonacoustic neuroma tumors (NANT) made up for the rest (52%). Meningiomas were the second more commonly diagnosed lesions, they constituted 32% of the cases. Meningiomas and AN were more frequent in women, their ratios being 7:1 and 1.6:1, respectively. In 60% of the cases the signs and symptoms became eloquent in patients of the fourth and fifth decades of life. CONCLUSIONS: The difference between our results and the ones previously reported in the medical literature are due in part to the predominance of female patients in our series. Endocrinologic, genetic and biochemical factors could also be responsible; nevertheless, this does not constitute the objective of the present study. PMID- 11939090 TI - The role of provocative discography in minimally invasive selective endoscopic discectomy. AB - A discogenic etiology is being widely accepted as a primary source of chronic low back pain. Even though increasing information is available on the pathophysiology of the degenerated disc, the present MR techniques are still not capable of pin pointing the source of this pain. In other words, with a non-invasive MR1 study we can still not define which disc is painful and what are the characteristics of discogenic pain from an imaging perspective. Discography remains therefore the only functional test that can delimit both morphologically and by provocation which discs are involved in the patents clinical picture. In combination with endoscopic procedures it helps tailor treatments. We consider that this diagnostic tool should be used routinely and performed by the surgeon himself as part of a screening-therapy protocol in spine surgery. We present our considerations regarding this diagnostic-screening method based on our clinical and surgical experience in patients who have been evaluated and have undergone selective endoscopic disscetomy after the screening. PMID- 11939091 TI - Surgical treatment of temporal lobe epilepsy. Personal experience. AB - The authors present their results with epilepsy surgery of the temporal lobe performed during a 5 years period at the Hospital of the University of Jordan, in 12 patients. All patients underwent comprehensive presurgical evaluation, including neurological history, examination, video-Electroencefalogram, Electroencephalogram for intercritical and habitual seizures and optimum magnetic resonance. After appropriate selection, we performed epilepsy surgery in 12 patients (9 males and 3 females). The mean age at operation was 21.4 years (range 4-44 years). All patients were receiving high therapeutic doses of antiepileptic therapy. Abnormal findings in Magnetic Resonance were observed in all cases. Pre and post resection electrocorticogram were recorded and graded according to Jay Scale. Pathological findings showed, neoplasic lesion in 2 cases (16%); one ganglioglioma and a low grade astrocytoma) and 10 patients (84%) showed non neoplasic lesions. Three patients (25%) had cortical dysplasia, another 3 (25%) had mesotemporal sclerosis, three more (25%) gliosis and one (8%) an arachnoid cyst. Post resection ECoG showed (84%) type A, (8%) type B, and (8%) type C. The present study has shown that intraoperative ECoG has an important role in the prediction of medically intractable epilepsy. Outcome of temporal lobe epilepsy surgery in our series was comparable to results reported by other authors despite of the absence of sophisticated functional tests. PMID- 11939092 TI - [Cerebellopontine angle lipoma: clinical case]. AB - Lipomas of the cerebellopontine angle are extremely rare. These tumors are probably maldevelopment lesions which can cause slowly progressive neurological symptoms. Including the present case, 90 lipomas in this localization have been described in the literature. The authors report a case of cerebellopontine angle lipoma in a 44-year-old male patient who suffered right hearing loss and tinnitus during seven months. The literature concerning this rare cerebellopontine angle tumor is review. The symptoms, radiological features and surgical management are discussed. PMID- 11939093 TI - [The entry of NEUROCIRUGIA in the Index Medicus/MEDLINE database]. PMID- 11939094 TI - [Primitive neuroectodermal tumor of the cauda equina. Report of a case]. AB - Primitive neuroectodermal tumors are malignant neoplasms whose classification has been controversial. Spinal primitive neuroectodermal tumors are rare and at the cauda equina are even less common. We report the case of a 40 year-old man with a history of progressive back pain and gait difficulty. After the diagnosis of a primitive neuroectodermal tumor of the cauda equina, the patient presented seeding to the intracranial subarachnoid space followed by a poor outcome. We review the literature of primary spinal primitive neuroectodermal tumors. PMID- 11939095 TI - [Primary low-grade non-Hodgkin's B lymphoma mimicking meningioma]. AB - Primary dural lymphomas are rare. Usually they present themselves as diffuse lesions and are infrequently well-delimited. We present a 44 year old male, who was admitted into our Centre after a generalised seizure. Neuroimage studies (CT scan, MRI and angiography) were performed, demonstrating a right hemispheric well delimited extra-axial mass, being diagnosed initially as meningioma. The patient underwent surgery with radical scission of the mass and dural margins. Histopathological diagnosis was Non-Hodgkin B-Cell lymphoma (marginal B-Cell lymphoma), which nowadays is considered as MALT lymphoma (Mucosa Associated Lymphoid Tissue). Studies directed to establish tumour extension didn't demonstrate the existence of another lesions in the organism. PMID- 11939096 TI - [Intradural cervical lipoma without neurologic involvement; report of a case]. AB - Lipomas are masses composed of mature adipose tissue, usually affecting lumbosacral levels, often associated with neural tube defects. Spinal lipomas at more rostral levels are usually unassociated with congenital abnormalities, but may produce a more severe neural compromise. Rare reports of cervical lipoma without neurological deficit have been described. We present the case of a 10 years-old girl who complained of cervical pain and stiff neck related to a C7-T1 intradural lipoma, without neurological affectation. The lipoma was partially resected without postoperative deterioration. PMID- 11939097 TI - [Comment on the article "Research training in neurosurgery residency programs. Analysis of the situation in other developed countries and proposals to complement the current program in Spain"]. PMID- 11939098 TI - [Intracranial purulent collections. Review of 34 surgically treated cases in 12 years (1989-2000)]. AB - We made a revision of 34 cases of intracerebral supurations (31 brain abscesses and 3 subdural empyemas) treated at our department of neurosurgery of "Mutua de Terrassa" during the period of 1989-2000. Treatment used was tapping of the abscess and aspiration in 28 cases and craneotomy and resection in only two cases. Three subdural empyemas were treated with burr-holes and aspiration. Results were evaluated using the Glasgow Outcome Scale at 6 months after discharge, resulting in 7 deaths (20.6%), 17 fully recovered patients (50%) and 10 with minor deficits (29.4%). Glasgow Coma Score at admission and the age were the unique variables significantly correlated with the final outcome. After analyzing the cost of treatment in our patients we suggest using antibiotic ambulatory treatment when the disease shows an adequate clinical and radiological response, thus permitting to shorten the usually long period of hospitalization of patients with intracerebral supurative diseases. PMID- 11939099 TI - [Award of the Spanish Society of Neurosurgery "Sixto Obrador, 2001"]. PMID- 11939100 TI - Computer-assisted generation of all-ceramic crowns and fixed partial dentures. AB - The successful application of the concept of computer-assisted manufacturing in restorative dentistry requires that computer-assisted design (CAD) and computer assisted machining (CAM) not only meet but actually exceed currently accepted standards for the material and clinical quality of dental restorations. In addition, the continued development of systems for polyvalent processing of disparate materials and objects must be assured. With these critical requirements in mind, the Precident system is a clinically proven and competitive system. The resolving power of the CCD chip is much improved compared to conventional cameras or charge-coupled devices. The scanner is able to scan entire casts in a fully automated process. In principle, this facilitates the production of frameworks for fixed prosthetic devices (FPD) of any size. It is also possible to create at least partial frameworks for removable prosthetic devices (RPD). A factor of great clinical and economic importance is the polyvalence of the process in materials processing: the numeric control (NC) machine can be programmed for metal alloys and ceramic materials as well as fiber-reinforced resins. At Aeskulap Klinik, the Precident System is routinely used for producing all single crowns and FPDs with up to four units. CAD/CAM all-ceramic crowns and FPDs currently cost about the same as metallo-ceramic or conventional all-ceramic restorations. PMID- 11939101 TI - Single-appointment, all-ceramic anterior restorations. PMID- 11939102 TI - Image compression in storing and communicating digital radiographs. PMID- 11939103 TI - Bar code, good for industry and trade--how does it benefit the dentist? AB - Every dentist who attentively follows the change in product labelling can easily see that the HIBC bar code is on the increase. In fact, according to information from FIDE/VDDI and ADE/BVD, the dental industry and trade are firmly resolved to apply the HIBC bar code to all products used internationally in dental practices. Why? Indeed, at first it looks like extra expense to additionally print a bar code on the packages. Good reasons can only lie in advantages which manufacturers and the trade expect from the HIBC bar code, Indications in dental technician circles are that the HIBC bar code is coming. If there are advantages, what are these, and can the dentist also profit from them? What does HIBC bar code mean and what items of interest does it include? What does bar code cost and does only one code exist? This is explained briefly, concentrating on the benefits bar code can bring for different users. PMID- 11939104 TI - Proximate and mineral compositions of cooked and uncooked Solanum melongena. AB - The proximate and mineral compositions of four different varieties of cooked and uncooked Solanum melongena were determined. Except for the percentage moisture, the levels of crude protein, crude fat, total ash and carbohydrate content were generally higher in uncooked samples than in cooked samples. Solanum macrocarpum (uncooked) has the highest moisture content with an average value of 73.9% and 84.5% for coat and flesh respectively, while the coat of Solanum sessiliflorum and flesh of Solanum aethiopicum have the least moisture content with average values of 60.0% and 69.5% respectively. The mineral components in the uncooked samples were generally found to be higher than those observed in the cooked samples. Solanum sessiliflorum has the highest value for nearly all the minerals for both the cooked and uncooked samples, while Solanum aethiopicum has the lowest value. The mineral contents were found to be higher in the coat than the flesh in most of the samples. Sodium and potassium were found to be the most abundant cations in both the coat and the flesh of cooked and uncooked fruits. PMID- 11939105 TI - Nutrient and sensory qualities of kunun zaki from different saccharification agents. AB - Kunun zaki--a cereal-based non-alcoholic, non-carbonated beverage--was studied. The ratio of blends of major ingredients, nutrient, amino acid content and sensory qualities of kunun zaki generated with different saccharifying agents were investigated. The main ingredients of the formulations were malted rice, sweet potato, soybeans and Cadaba farinosa (Dangarafa or Legel in Hausa), each used separately with sorghum to produce a kunun zaki type. The weight ratios of the major ingredients were 8:91 for malted rice-sorghum, 7:92 for sweet potato sorghum, 9:90 for soybean-sorghum and 4:95 for Cadaba farinosa-sorghum blends with ginger contributing 1% in each case as a spice. The nutrient composition of kunun zaki samples from different saccharifying agents ranged from 87 to 91% for moisture, 3.19 to 7.86% for crude protein, 0.37 to 0.75% for fat, 0.93 to 1.20% for ash and 2.69 to 5.84% for carbohydrate. Glutamic acid (4.49-11.66 g) was the most abundant amino acid in the samples while cysteine was the least abundant (0.34-1.45 g) in all the samples. The lowest concentration of all the essential amino acids except for tryptophan occurred when malted rice was used (0.44-1.40 g). Among the essential amino acids, cysteine, valine, isoleucine and methionine occurred in extremely low quantities compared with FAO/WHO reference protein values. The dual role (saccharification and enrichment) of soybean in kunun zaki processing is a desirable attribute and offers an advantage over the other agents. The different saccharifying agents had no significant effect (P > 0.05) on colour and flavour of kunun zaki but did influence sweetness, mouthfeel and overall acceptability. The beverage made with malted rice was most liked overall. PMID- 11939106 TI - Mineral content of culinary and medicinal plants cultivated by Hmong refugees living in Sacramento, California. AB - Since the end of the American-Vietnamese War in 1975, more than 1.5 million refugees from Southeast Asia have resettled in the United States. Included among these displaced persons were the Hmong from Laos, a subsistence-based, shifting cultivation, agricultural society. Hmong who resettled in urban areas have viewed vacant lots adjacent to urban dwellings as potential garden sites for production of familiar herbs and vegetables. In the present study exotic culinary and medicinal herbs grown by Hmong refugees in Sacramento, California were identified and analyzed for mineral composition. The herbs grown in these urban gardens were significant ingredients of Hmong recipes, and herb leaves, or infusions of steamed herb leaves were widely consumed as a component of pregnancy and post partum diets. Six common species, Acorus gramineus, aff. Angelica, Dendranthema indicum, Eupatorium lindleyana, Sedum aff. sarmentosum, and Sedum aff. spectabile, were used in combination to season chicken. Polygonum odoratum, also widely cultivated, was used to season fish. Exotic culinary-medicinal species with highest mineral profiles included: Basella alba (Ca, Mg, Mn, Zn); Houttuynia cordata (Fe, Mg, Mn); Justica gendarussa (Ca, Mg, Zn); and Polygonum odoratum (Ca, Mg, Mn). While vacant lots sometimes are heavy metal contamination sites, we found no detectable levels of arsenic, cadmium, chromium, or lead in the samples analyzed. PMID- 11939107 TI - Development and validation of a food frequency questionnaire for Korean Americans. AB - A food frequency questionnaire for Korean Americans (KFFQ) was developed by modifying Block's Health Habits and History Questionnaire (HHHQ). The final KFFQ contained 118 food items taking into account culturally specific dietary patterns. Validation of the KFFQ was conducted to compare the nutrient intakes assessed by the KFFQ against those measured by seven-day dietary records in 73 Korean Americans. The mean nutrient values from the KFFQ differed at most by 25% from those of the dietary records with the exception of vitamin A, and 70% of the nutrients (16 out of 23 nutrients) were within 15%. Adjusted Spearman rank-order correlation coefficients for nutrients ranged between 0.45 and 0.84. The overall degree of agreement between the KFFQ and the dietary records was 68% with a range 56% to 79%. This validation study indicates that the KFFQ gives reasonably accurate estimates of the usual dietary intakes among the study population. The results also verify that it is possible to use a FFQ that is both culturally specific and comprehensive with the benefit of being easy to self-administer. This high degree of correlation between the KFFQ and the dietary records demonstrates that the KFFQ provides an easier yet accurate method in large-scale epidemiological studies of relationships between nutrition intakes and diseases in Korean Americans. PMID- 11939108 TI - The nutritional quality of an infant food from quinoa and its effect on the plasma level of insulin-like growth factor-1 (IGF-1) in undernourished children. AB - An infant food product was manufactured by drum drying a pre-cooked slurry of quinoa (Chenopodium quinoa, Willd) flour. The chemical composition shows that the product is a potential source of valuable nutrients, like protein (16%), vitamin E (19 mg/kg), thiamine (0.7 mg/100 g), iron (70 mg/kg), zinc (48 mg/kg) and magnesium (1.8 g/kg), all the values expressed on dry basis, to pre-school children (of 5 years of age). The animal feeding experiments with rats showed a net protein utilisation (NPU) of 68, digestibility (TD) 95 and biological value (BV) 71. The level of insulin-like growth factor-1 (IGF-1) in the plasma of the children who consumed a supplementary portion of 2 x 100 g of the above infant food product showed an increase after a period of 15 days, while the plasma level of IGF-1 in the children of the control group as well as the reference group did not show any significant increase. PMID- 11939109 TI - Physical properties and composition of femurs of rat fed with diets based on corn tortillas made from different processes. AB - In this study the effect of calcium absorption on some physical properties and composition of rat femurs was evaluated, comparing rats fed with raw whole corn (RC), tortillas made from extruded masa with 0.25% lime content (TEWL) and without lime (TE), and nixtamal tortillas (NT). The diets were formulated to contain the same amount of protein, oil, fiber, vitamins and minerals other than calcium. In all diets 0.20% calcium was added. At the end of the trials, the femurs were extracted, weighed and measured for ash, calcium and phosphorus content, some physical dimensions, and the crystallinity percentage. The femurs of rats fed with TEWL and NT were heavier, thicker, longer and had higher calcium content. On the other hand, the force required to break the femur of rats fed on ETWL and NT was 1.25 kg greater than that required to break the femurs of rats fed with RC. Higher crystallinity percentage values were observed in the femurs of the rats fed with NT (37.66%) and TEWL (36.98%) as compared to a 30.31% value obtained with the RC. PMID- 11939110 TI - An evaluation of two Mexican food guides. AB - In Mexico, the food guide developed by The National Institute of Nutrition for the Mexican population is 'The Pyramid of Health'. In the northwestern Mexican state of Baja California, the Nutrition Institute of Baja California (INUBAC) developed and validated another food guide. This guide, named 'The Apple of Health', is based on current nutrition research and the typical food patterns. The purpose of this article was to evaluate and compare (1) the graphical impact, (2) the understanding of the overall message, and (3) the ability to apply the information from both food guides, using a diet design score and focus group interviews, among women from two different education levels living in Baja California. Results showed that mean diet design percentage scores obtained by the participants using The Pyramid of Health were 64.1 and using The Apple of Health 76.7. No statistical difference was found between the two education levels. The majority of the focus group participants expressed that the most appealing food guide was The Apple of Health because of its color and graphic art. However, the Pyramid was more familiar to the participants. The study subjects strongly recommended The Apple of Health food guide as an educational tool for the whole family. PMID- 11939111 TI - Availability of vitamin B6 from different food sources. AB - The present study was conducted to investigate the precaecal digestibility of vitamin B6 from selected food sources including eggs, bananas, white cabbage, corn, milk powder, fish, barley, soybeans, brown rice, wheat bran, brewer's yeast, rye and soybean meal. These foods were chosen for their relevance in human and animal nutrition and prepared as they are usually eaten by humans. As confirmed by further investigations the precaecal digestibility is a valuable measure for determining the availability of native B-vitamins. Therefore, pigs were fitted with an end-to-end ileo-rectal anastomosis for digesta passing straight from ileum to rectum, thus avoiding endogenous vitamin synthesis by the colon. Three weeks after surgery the digestibility experiments were carried out during which the animals were fed various experimental diets for a period of 12 days and digesta were collected quantitatively twice a day during the final 5 days of this period. The concentration of vitamin B6 in foods and chyme was determined by high pressure liquid chromatography. Precaecal digestibility of vitamin B6 from all tested food sources ranged from 51 to 91% in the following order: cabbage diet > banana diet > fish diet > milk powder diet > brewer's yeast diet > soybeans diet > soybean meal diet > egg diet/corn diet > barley diet > wheat bran diet > rye diet. Only boiled brown rice had a very low vitamin B6 availability of 16%. The digestibility of vitamin B6 from plant products (excluding the rice) was on average 10% lower when compared with animal products (71 versus 79%). PMID- 11939112 TI - Simple physical treatment as an effective tool to improve the functional properties or rapeseed (Brassica campestris var. toria) and sesame seed (Sesamum indicum) meals. AB - Dry and 24 h imbibed rapeseeds and sesame seeds were defatted with hexane and the resulting freeze-dried powder was analysed for the functional properties of the meals. Water absorption capacity (WAC) of imbibed rapeseed meal and fat absorption capacity (FAC) of both the imbeded meals were higher than those for dry meals. Protein solubility of rapeseed meals was improved by imbibition and both the imbibed meals exhibited maximum protein solubility at pH 12. Rapeseed meal possessed better foaming properties and viscosity than sesame seed meal. Imbibition considerably enhanced the foaming properties of rapeseed meal while the emulsification properties and viscosity did not change appreciably. Emulsification properties of sesame meal were higher than rapeseed meal. PMID- 11939113 TI - [Cultural adaptation of instruments utilized in occupational health]. AB - Numerous standardized questionnaires and other instruments for evaluating health status and treatment effectiveness have been developed. However, the majority of these instruments have been constructed and validated in English. The scientific literature generally recommends the use of scales and standardized questionnaires whose results can be compared internationally. However, because of cultural differences, it is not sufficient to just do a simple translation of these instruments. Drawing on specialized research in these area, this article presents a summary of systematized standards for the cultural adaptation of instruments used in occupational health. PMID- 11939114 TI - [Fatal deception: the tobacco industry's "new" world standards for tobacco marketing]. AB - Being increasingly threatened by the worldwide antismoking struggle, the major tobacco companies are eager to improve their public image. This leads the companies to adopt inconsequential "measures" such as the tobacco industry's "new" standards for tobacco marketing that were "voluntarily" issued in September 2001 by the British American Tobacco company. These measures are clearly attempts to reduce the disapproval generated by the companies' promotion and advertising campaigns, which indirectly target young people. With these standards the tobacco companies supposedly commit themselves, among other things, to not using advertising directed at youth and to not selling or distributing tobacco products in places frequented by young people. This document explains why these measures are completely ineffective, are not anything new, and are a subtle effort to feign a conscientious, responsible attitude, which is far from genuine. As long as there are marketing activities directed at adults, young people will be exposed to the influence of those activities. Many countries have completely prohibited the marketing of tobacco products, given that the "new" marketing standards do not represent progress in any way whatsoever. PMID- 11939115 TI - Improving public health by more effectively regulating tobacco products. PMID- 11939116 TI - [A possible world]. PMID- 11939117 TI - [Three fundamental measures to revitalize the control of smoking in the Americas]. PMID- 11939119 TI - [Design and implementation of an epidemiological surveillance system for mental health]. AB - OBJECTIVE: To design and test a surveillance system to detect and rank the most significant mental health problems in a community and to establish their association with the living conditions in that community. METHODS: The system was designed and tested at a primary health care center in the community of La Florida, which is in the metropolitan region of Santiago, the capital of Chile. The health problems that were under surveillance, with monthly monitoring, were: addiction (to alcohol, tobacco, and benzodiazepines or other drugs), violence (domestic violence and sexual abuse, real or suspected), and disorders in children (attention deficit and behavioral disorders). By means of a computerized geographical information system (GIS), those surveillance data were combined with other data. These other data included sociodemographic information on the persons who went to the health center for care as well as data on local risk factors and protective factors for health problems, some of which are indicators of the local residents' living conditions. RESULTS: The proposed epidemiological surveillance system, which takes into account local living conditions and environmental variables, is a good tool for health action. The proposed system also complements the incomplete perspective of monitoring systems, which do not include variables describing the local context and that focus only on health problems. The information synthesized in the GIS maps makes it possible to simultaneously display different layers of information on factors related to the health problems studied, linked to a specific area and all its environmental variables, in an easy-to-read, self-explanatory format. CONCLUSIONS: These results and the concurring opinions of the participating physicians show that the experimental system effectively met the basic requirements of an epidemiological surveillance system of this kind. PMID- 11939118 TI - Behavioral problems and tobacco use among adolescents in Central America and the Dominican Republic. AB - OBJECTIVES: The purpose of this study was to examine the association between behavioral problems and tobacco use among adolescent students in six countries of Central America and in the Dominican Republic. METHODS: Data were drawn from a multinational collaborative study that included questionnaire surveys of between 451 and 1,170 school-attending adolescents in each of the seven countries studied. Assessments were based on an adapted, Spanish-language version of the Drug Use Screening Inventory (DUSI). The conditional form of logistic regression was employed for analysis, matching students on type of school and area, with further statistical adjustments for sex, age, and selected risk factors. RESULTS: Occurrence of tobacco use was observed to vary dramatically from country to country. Nonetheless, for the combined group of countries, the estimated odds of tobacco use in youths at the highest levels of behavioral problems was more than five times that for youths at the lowest levels, after controlling for sex, age, lack of participation in recreational activities, level of irritability, and levels of problems with school, family, and mental health. Country-specific analyses show that youths at the highest levels of behavioral problems have a consistently greater occurrence of tobacco use as compared to youths at the lowest levels of behavioral problems. CONCLUSIONS: These findings are concordant with prior studies on tobacco use among adolescents with behavioral problems. Although the magnitude of observed associations varied according to the country of residence, the strength of these associations and their significance by conventional standards were observed in nearly all the countries sampled. This is the first study in these seven countries on potentially causal relationships such as these. More research is needed to augment our knowledge regarding the observed cross-country differences and ultimately to develop, implement, and evaluate effective tobacco preventive intervention programs. PMID- 11939120 TI - [Enterobacteria isolated from cockroaches (Periplaneta americana) captured in a Brazilian hospital]. AB - OBJECTIVE: To isolate and identify microorganisms from cockroaches that were captured in a public hospital and to test the antimicrobial susceptibility of these microorganisms. METHODS: Cockroaches were captured in the morning and at night. They were placed in flasks rinsed with 70% alcohol, transferred to sterilized flasks, and then taken to the laboratory. Only cockroaches captured whole and live were utilized for the study. After being immobilized at 0 degree C, each cockroach was placed in a test tube with sterile saline solution (0.8%) and then homogenized. The resulting solution was then placed in the following five culture media: MacConkey agar, nutrient broth, brain-heart infusion agar, Sabouraud agar, and mannitol. The cultures were examined using a stereomicroscope, and colony-forming units were counted. The disk diffusion test was used to determine antimicrobial susceptibility. RESULTS: We found a 56% prevalence of enterobacteria and an 18% prevalence of coagulase-negative staphylococci. Fifteen species of enterobacteria were identified. The most frequent were Klebsiella pneumoniae (17%), Enterobacter aerogenes (14%), Serratia marcescens (13%), Hafnia alvei (12%), Enterobacter gergoviae and Enterobacter cloacae (each 9%), and Serratia spp. (6%). Both the enterobacteria and the coagulase-negative staphylococci showed significant resistance to antimicrobials, including oxacillin. CONCLUSIONS: The prevalence of enteropathogenic bacteria and coagulase-negative staphylococci isolated from Periplaneta americana cockroaches in the studied hospital reflects the weakness of the measures adopted both for vector control and for antimicrobial use. The results show the need to implement effective health-institution programs focusing on hygiene and the rational use of antimicrobials. PMID- 11939121 TI - Peruvian mothers' knowledge and recognition of pneumonia in children under 5 years of age. AB - OBJECTIVE: To assess Peruvian mothers' knowledge and recognition of pneumonia in children under 5 years of age, the mothers' attitude toward seeking medical help if they had a child with signs of pneumonia, and their perception of a Government of Peru pneumonia campaign. METHODS: In this cross-sectional study, 501 mothers were selected randomly from 20 low-income communities of the metropolitan area of Lima, Peru, and were interviewed between June and August 2000. Using SPSS software, descriptive statistics were applied to summarize the demographic data and the data regarding the mothers' knowledge of pneumonia and recognition of signs of the disease. Cross-tabulations and chi-squares were done to assess relationships between variables and to make comparisons. RESULTS: About 84% of the mothers said that they knew what pneumonia is. Most believed that pneumonia is dangerous. A majority (58.7%) indicated that pneumonia is caused by lack of parental care. Only 28.9% believed that a virus causes the disease. More than 80% correctly picked rapid breathing and/or chest retraction from a list of possible signs and symptoms of pneumonia, and 94.6% said they were ready to take their child to the closest health center if they thought their child had pneumonia. Although 57.1% said they had heard about the Government of Peru pneumonia campaign, 69.3% of these mothers said they could not recall the motto of the campaign. Mothers who reported having heard of the campaign through TV were more likely than other mothers to correctly recognize the two major signs of pneumonia presented in the campaign. CONCLUSIONS: Although the percentage of mothers believing they can recognize pneumonia through rapid breathing and chest retraction seems to have increased in recent years, there is still a sizable percentage of mothers who remain uninformed about pneumonia and its possible fatal consequences. Efforts need to continue to educate Peruvian mothers about the causes, recognition of the signs, and treatment of pneumonia. The results suggest that the Government of Peru pneumonia campaign should use television much more, as well as the health centers, where most of the mothers receive medical attention and health information. PMID- 11939122 TI - [Patients in treatment for malnutrition in primary care, study of 500 real patients]. AB - OBJECTIVE: The worsening of the nutritional status of certain segments of the population has led to frequent situations of chronic undernourishment even in the healthy population. There are very few data available on the prevalence and causes of malnutrition in Primary Health Care. The present study attempts to provide measurable information, obtained at random from the doctors involved in the country's Primary Health Care, on the characteristics of the undernourished patients, the cause of the undernourishment, the diagnostic means used, the treatment applied and the progress of the patients regularly treated in Primary Health Care facilities. MATERIALS AND METHOD: A sample of 1,819 doctors in Primary Health Care were surveyed to know their opinions on the nutritional status of their patients. They were asked to complete a "Patient Record" for the first patient to enter their office suffering from undernourishment. A total of 505 Patient Records were received from the different Primary Health Care doctors taking part in the study throughout Spain. RESULTS: Of the patients included, 10% were aged less than 10, while 46.7% were between 16 and 65 years of age and 44.2% were over 65. The main diagnosis in these patients was varied, with cancer patients (22.6%) and anorexics, including anorexia nerviosa and other non oncological causes, (16.4%) the most common. As for the tests used for diagnosing undernourishment, those most frequently applied were physical examination (61%) and biochemical tests (56.4%). The risk factor most commonly found in these patients was old age/senility (21%). Nutritional support (55.8%) and dietary recommendations (45.3%) were the therapies most often applied. Only 47% of patients correctly implemented their treatment according to the doctors in Primary Health Care and the prognosis was as follows: 31% were expected to improve, 20% to worsen and 44% of cases would remain stable. CONCLUSIONS: From this study, it is concluded that most undernourished patients in Primary Health Care are there due to a severe pathology or because of age; that anthropometric and biochemical methods are used for diagnosis purposes although the clinical interview is of basic importance; that a large proportion of patients require some type of nutritional supplements; and finally that, according to the doctors, the expectations of improvement in the nutritional status of these patients are not good. PMID- 11939123 TI - [Long-term follow-up of home parenteral nutrition at a general hospital: complications and quality of life]. AB - OBJECTIVE: The present study assesses the septic, thrombotic and hepatic complications and also the quality of life of patients with home parenteral nutrition (NPD in its Spanish acronym). METHODS: The case histories of patients treated with NPD at our hospital were collected for the period January, 1986, to January, 2001. We had 20 patients, mean age 48 +/- 14 years (mean +/- SD), with a total catheterization time of 16,292 days. The main causes of intestinal failure were mesenteric ischaemia and radical enteritis. Quality of life was assessed by means of the Sickness Impact Profile (SIP) and the Karnofsky Index (KI). RESULTS: We found a catheter infection rate of 0.42 per 100 days of catheter, mostly produced by coagulase-negative staphylococci (63%). In 30% of the cases, it was necessary to remove the catheter, including in all cases produced by Candida. Cather occlusion rate was 0.05 per 100 days. Five patients presented an increase in transaminases and alkaline phosphatase, and one female patient developed cholestasis. Two patients died of NPD-related causes. The results of the quality of life analysis showed it was moderately affected, with SIP scores of 14.6 +/- 3.5 and KI results of 70 +/- 20 (mean +/- SD). CONCLUSIONS: Home parenteral nutrition allows a greater survival in cases of intestinal failure. The most frequent complication is an infection of the catheter. The results of the quality of life analysis in our series reveal it was moderately affected in most cases. PMID- 11939124 TI - [Glycine: a cell-protecting anti-oxidant nutrient]. AB - For many researchers it is still difficult to accept that beneficial effects can be obtained in several disease states with the simplest amino acid, glycine. However, evidence is mounting in favour of this idea. It is now clear that dietary glycine protects against shock caused either by blood loss or endotoxin, reduces alcohol levels in the stomach and improves recovery from alcoholic hepatitis, diminishes liver injury caused by hepatotoxic drugs and blocks programmed cell death and reduces the nephrotoxicity caused by the drug cyclosporin A in the kidney, preventing hypoxia and free radical formation. It could be also useful in other inflammatory diseases since it diminishes cytokines production. We review some of the beneficial effects of glycine and their responsible mechanism, which could led to advice its use in the therapy of different diseases. PMID- 11939125 TI - [Detection of malnutrition risk in hospitalized elderly patients]. AB - OBJECTIVE AND SCOPE: To validate a protocol for the detection of malnutrition risk in a population of elderly patients admitted to a general hospital for non surgical reasons. MATERIALS AND METHOD: The study involved 95 patients (34 of them male) over the age of 65 who were assessed on the third and fifth day after admission by means of a simple screening protocol (PC in its Spanish acronym) that considered recent changes in weight, serum albumin, lymphocyte concentration, food intake and diagnosis on admission, together with a more complex diagnostic protocol (PD in its Spanish acronym) including anthropometric, biochemical and immunological parameters. The PC was applied by personnel not expert in nutrition, while the PD was carried out by persons trained in nutritional assessment. The results of the PC and PD were compared, with statistical significance being considered at levels of p < 0.05. RESULTS: According to the PD, 75 patients (78.9%) suffered protein-energy malnutrition (39 slight, 31 moderate, 5 severe). The score obtained in the PC had a significant relationship with the severity of the malnutrition diagnosed using PD (p < 0.001). Furthermore, the absence or presence of nutritional risk assessed using the PC had a significant correlation with the diagnosis of malnutrition and its degree according to the PD (p < 0.01), thus indicating the validity of the PC as a method for screening of malnutrition. CONCLUSIONS: 1. In our population of geriatric patients admitted for reasons other than surgery, there was a high prevalence of protein-energy malnutrition. 2. The risk of malnutrition in an elderly population in hospital settings is appropriately assessed by means of a simple screening test effected by non-specialized personnel. 3. The score obtained in the PC is linked with the presence of malnutrition and with its severity. 4. The use of simple screening techniques in populations with a high prevalence of malnutrition may contribute to the detection and correction of this problem. PMID- 11939127 TI - [Evaluation of Hospital Nutrition III. Delay, subjects and citation]. AB - We continue with the bibliometric analysis of Nutricion Hospitalaria, in this case providing the data on lag times for the publication of articles, analysis of subject matter, references and citations, including also the calculations of the impact factor, an essential factor for the inclusion of any publication in Current Contents. PMID- 11939126 TI - [Home enteral nutrition. Annual report 1999. NADYA-SENPE Group]. AB - GOAL: During 1999, as in previous years, the NADYA-SENPE Group has maintained an annual register of patients with Artificial Nutrition at Home in order to keep up to date our available knowledge of this therapy. The present paper analyzes the results of the sixth National Register of patients under treatment with Enteral Nutrition at Home corresponding to 1999, produced with the co-operation of twenty three centres in the Spanish national health network. MATERIAL AND METHODS: The data were collected through a closed questionnaire included on our web site (www.nadya-senpe.com). Apart from epidemiological information, the form includes the indication that led to the prescription of nutrition, nutritional treatment, access path, complications and admissions to hospital, follow-up of the treatment, patients' quality of life and progress. All of the data are processed by the co-ordinating team. The Nutrition Unit at La Paz Teaching Hospital in Madrid has acted as the group co-ordinator. RESULTS: During 1999, a total of 2,262 patients at the twenty-three collaborating centres followed treatment with Home Enteral Nutrition (NED in its Spanish acronym). The mean age was 63.6 (19.67 years (males: 57.6%; females: 42.3%). The mean time with nutritional treatment is 5.89 (4.25 months. The neurological alterations (37.5%) and neoplasias (36.8%) were the most frequent indications for NED. Most patients used oral administration (50.7%), the use of artificial routes is less frequent, with 5NG being used on 27.9% and PEG on 12.7%. The polymeric formulas are the ones most commonly used (87.7%). The number of complications recorded amounted to 1,403 episodes, representing 0.62 complications per patient per year, of which 40.8% were gastro-intestinal (0.26 complications per patient per year) and 18.7% were mechanical complications, with only 9 recorded cases of bronchoaspiration. It was necessary to admit patients to hospital on 836 occasions (0.38 admissions per patient), albeit generally for causes not associated with nutrition (86.1%). At year-end, 51.4% of patients continued to receive nutritional support. CONCLUSIONS: Enteral nutrition at home is a safe technique whose use is growing in Spain. It requires the existence of appropriately equipped nutrition teams as well as the collaboration of the Primary Health Care services and the suitable training of patients and relatives. It is essential to progress more deeply into the development and application of clinical routes that provide a clear and concise methodology covering all nutrition-related aspects. In itself, nutrition is a therapeutic alternative regardless of whether access is obtained through a natural or an artificial route. It is necessary for the persons responsible for health-care administration to realize that the appropriate nutrition of patients leads to an improvement in their general health, a shorter course for their illness and, therefore, a reduction in costs. PMID- 11939128 TI - [Nutritional support in chylothorax secondary to lymphoma]. AB - Chylothorax is a well-known, albeit infrequent, complication in certain neoplasias including lymphomas. The continuing loss of chyle leads to a profound deterioration in patients' nutritional and immunological status, prevented only by appropriate early nutritional support. There is currently some dispute over its handling, particularly with regard to the most appropriate type of nutritional support and the suitability of conservative treatment versus surgery. The present paper describes the case of a 41-year-old patient diagnosed as having secondary chylothorax following B-cell lymphoma where mixed nutritional support (fat-free enteral nutrition by mouth and total parenteral nutrition) was given, followed by talc pleurodesis, with optimal results. There is a review of the literature and a discussion of the most controversial aspects of its management. PMID- 11939130 TI - [Knowledge and attitude on deviation in medical clinical practice]. PMID- 11939129 TI - [Sinus bradycardia in anorexia nervosa]. AB - Anorexia nervosa is a frequent condition of malnutrition in industrialized countries. The most frequent causes of death in patients with anorexia nervosa are suicide and sudden death generally in relationship with cardiovascular abnormalities. We report a case of a 42-year-old woman with anorexia nervosa who was admitted to our hospital because of severe weight loss. Her electrocardiogram showed a severe sinus bradycardia of 27 b/min. PMID- 11939131 TI - [Letter to the editor from the authors Dr. Pilar Cervera, Dr. Jaume Clapes and Dr. Rita Rigolfas]. PMID- 11939132 TI - [Francis Daniels Moore in 1975 (1913-2001)]. PMID- 11939133 TI - Pharmacogenetics research network and knowledge base: 1st annual scientific meeting. AB - The National Institute of General Medical Sciences of the National Institutes of Health recently established a collaborative group of scientists, called the Pharmacogenetics Research Network. Central to the network is a shared, state-of the-art data repository, the Pharmacogenetics Knowledge Base (PharmGKB), which is housed at Stanford University. Network investigators deposit pharmacogenetic data into PharmGKB, after all individually identifying information has been removed. Contents of PharmGKB will be freely accessible to the scientific community, with the goal of forging new links between gene variation and drug response. An open scientific meeting was held recently to introduce the research community to the network and to invite academic and industry-based researchers to deposit data into PharmGKB. Featured at the meeting were summaries of research progress to date, as well as discussions of issues intimately related to pharmacogenetics research, namely ethics and relations with the biotechnology and pharmaceutical industries. PMID- 11939134 TI - 4th Annual Pharmacogenomics and Medicine Lectures. AB - In the future, pharmacogenomics will play an important role in the treatment of patients by making it possible to predict drug response based on an individual's genetic make-up. Similarly, pharmacogenomics may be used to reduce the probability that adverse effects will occur. The use of a patient's genetic information will lead to greater predictability in clinical outcomes and personalisation of medical care. Pharmacogenomic information can also aid in drug development by helping to select individuals that are likely to respond to a medication for participation in clinical trials. Integration of pharmacogenomics into the healthcare system has a number of potential economic benefits, including reduced costs of healthcare and drug discovery. The FDA has no specific plans to regulate therapy-guiding pharmacogenomic tests, which are different from diagnostic genetic tests. There are a number of ethical issues related to pharmacogenomics, including the credibility of the system for protecting the rights and welfare of human research subjects, general concerns about genetic research, privacy issues and equitable distribution of the technology. To ensure integration of pharmacogenomics into the healthcare system it will be important to obtain public support through education about the benefits and risks of this technology. PMID- 11939135 TI - Cambridge Healthtech Institute's 5th Annual Conference: impact of genomics on medicine. AB - The recent publications in Nature and Science by the Human Genome Consortium and Celera Genomics, respectively, while being landmark achievements in themselves, have also given pause for thought. A definitive catalogue of human genes is still not available but the broad picture of how humans compare with lower organisms at the genomic level is becoming clearer. The full impact of these findings on the practice of medicine is hard to predict, but research being conducted now, in the early years of the 21st century, will form the basis of future advances in the diagnosis and treatment of disease. Exactly what this will entail is the subject of intense debate, but there are some common starting points that were discussed at this meeting in Munich. The main theme to emerge was the need to move beyond the human genome sequence towards an understanding of proteins and their interactions in complex biological pathways, thereby increasing opportunities for drug discovery through the identification of new targets. The majority of the talks were therefore devoted to the description of technological advances in the analysis of gene and protein expression (and interaction) and in the use of various methods of gene deletion in order to validate individual proteins as drug targets. Perhaps it will still be a few years before it will be possible to report on the application of genomic analyses to routine medical practice at the first point of care for patients but when that happens, the research efforts described here will have been worthwhile. PMID- 11939137 TI - Proceedings of the 4th World Conference on Early Pregnancy Continuum Between Implantation and Perinatal Events. Pecs, Hungary, June 1-3, 2000. PMID- 11939136 TI - Cambridge Healthtech Institute's 2nd Annual Conference on Pharmacogenomics Europe: presaging profits. AB - Pharmacogenomics promises to offer distinct strategic advantages to pharmaceutical companies, physicians, providers and patients. Cambridge Healthtech Institute's 2nd Annual Conference on 'Pharmacogenomics Europe: Presaging Profits' covered all aspects of pharmacogenomics and gave scientists from both academia and from pharmaceutical and biotech companies a great opportunity to discuss the latest progress in pharmacogenomic research. The meeting considered technologies for single nucleotide polymorphism (SNP) screening and expression profiling, bioinformatic tools for data evaluation and gave an overview on the state of affairs and novel approaches to implement pharmacogenomics and pharmacogenetics into drug development and medical treatment. The major strength of the meeting was the merging of scientists from many different disciplines, such as clinicians, pharmacologists, molecular biologists, engineers and bioinformatics experts, into one meeting. PMID- 11939138 TI - The first description of the eye muscle pulleys by Philibert C. Sappey (1888). PMID- 11939139 TI - The motor muscles of the eyeball [translation from the French]. 1888. PMID- 11939140 TI - Fourth International Symposium on the Role of Soy in Preventing and Treating Chronic Disease. San Diego, California, USA. November 4-7, 2001. Proceedings and abstracts. PMID- 11939141 TI - The pepsinogen releasing effect of Helicobacter pylori lipopolysaccharide. AB - BACKGROUND: Helicobacter pylori lipopolysaccharide (LPS) affects pepsinogen release by a nontoxic mechanism. We hypothesized that this effect was characteristic of the organism and related to the clinical status of the strain. MATERIALS AND METHODS: LPS was isolated from 11 H. pylori strains whose pathogenic profile was known and four other nongastric bacteria. The effects of luminal LPS on guinea pig gastric mucosal pepsinogen release was evaluated using the Ussing chamber technique. CCK-8 (10(-9)M) was used as a positive control. RESULTS: H. pylori LPS dose-dependently stimulated pepsinogen release with a maximal stimulation at 250 microg/ml (approximately 4500; p < .001 vs. control). LPS from other Helicobacter or Campylobacter species had no effect on pepsinogen release. ANOVA demonstrated significant differences in the efficacies of pepsinogen release between the 11 clinical H. pylori strains (p < .0001) despite the fact that they were all cagA+ and 90 had the cytotoxic vacA subtype s1. Physical and chemical disruption of the LPS suggested that both the structure and the carbohydrate composition of this molecule may play a critical role in pepsinogen release. Polymyxin B partly (p < .03) inhibited and dephosphorylation completely inhibited (p = .0002) LPS-stimulated pepsinogen release. CONCLUSION: Pepsinogen release is an innate property of all cagA+H. pylori LPS. The structure of the molecule and composition of side-chains are important in this response which appears to be partially lipid A driven. PMID- 11939142 TI - [Ecos Internacionales. 16-18 August 2001, Mexico City, Mexico]. PMID- 11939143 TI - Canadian Dermatology Association membership directory 2002. PMID- 11939144 TI - Abstracts of the 2002 annual convention of the American Alliance for Health, Physical Education, Recreation and Dance. San Diego, California, USA. April 9-13, 2002. PMID- 11939145 TI - Abstracts of the 33rd Annual Meeting of the Environmental Mutagen Society. April 27-May 2, 2002. Anchorage, Alaska. PMID- 11939146 TI - Abstracts of the 34th Annual Meeting of the American Burn Association. April 24 27, 2002. Chicago, Illinois, USA. PMID- 11939147 TI - Abstracts of the 7th Meeting of the European Society of Gene Therapy (ESGT). Munich, Germany, 26-28 November 1999. PMID- 11939148 TI - Abstracts of the 6th Meeting of the European Working Group on Human Gene Transfer and Therapy (EWGT). Jerusalem, Israel, 21-24 November 1998. PMID- 11939149 TI - Abstracts of the 71st Annual Meeting of the American Association of Physical Anthropologists. Buffalo, New York, USA. April 10-13, 2002. PMID- 11939150 TI - [Bochum 2002. 43rd Congress of the German Society of Pneumology and 22nd Meeting of the German Society of Allergy and Clinical Immunology. 13-16 March 2002. Abstracts]. PMID- 11939151 TI - European Congress of Radiology. Vienna, Austria, March 1-5, 2002. Abstracts. PMID- 11939152 TI - Orthopaedic Proceedings 2000, 2001. PMID- 11939153 TI - EBMT 2002. 28th Meeting of Physicians, 18th Meeting of the Nurses Group, 1st Meeting of Data Management Group. Montreux, Switzerland, March 24-27, 2002. Abstracts. PMID- 11939154 TI - Abstracts from the Society of Cardiovascular Anesthesiologists 24th annual meeting. New York, New York, USA. April 20-24, 2002. PMID- 11939155 TI - The regulation of the transition from research to clinical practice in human assisted conception. PMID- 11939156 TI - Optimizing gonadotrophin-releasing hormone antagonist protocols. AB - The introduction of the gonadotrophin-releasing hormone(GnRH) antagonists offers new potential for in vitro fertilization (IVF) clinics. Compared with GnRH agonists, the use of the GnRH antagonists significantly reduces the dose of gonadotrophin and duration of treatment required, and also reduces unwanted side effects. Patients also tend to prefer treatment with GnRH antagonists compared with agonists. The GnRH antagonists are useful in both good and poor responders, and there is some flexibility in treatment protocols. A single dose of GnRH antagonist may be used in patients who do not want or require more aggressive stimulation. Promising data indicate advantages of GnRH antagonists in terms of reduced incidence of ovarian hyperstimulation syndrome and reduced impairment of luteal function. It is anticipated that, as a result of further development of treatment protocols, pregnancy rates with the GnRH antagonists will become at least equivalent to those achieved with GnRH agonist protocols. PMID- 11939157 TI - Monitoring during gonadotrophin-releasing hormone antagonist protocols. AB - The results of the four large industry sponsored multicentre clinical trials comparing gonadotrophin-releasing hormone (GnRH) antagonists and agonists in controlled cycles of assisted reproduction are reviewed from the perspective of implications for monitoring ovarian responses. The simplicity and consistency of antagonist-controlled cycles in normal women indicate that patients in assisted reproduction programmes should undergo fewer assessments and will receive shorter treatment cycles. However, the two-stage pattern of follicular recruitment in antagonist-controlled cycles leads to follicles of different size profiles at the end of the follicular phase compared with those obtained in agonist-controlled cycles (one recruitment process only). Increasing the dose of follicle stimulating hormone (FSH) may increase oocyte yield, but does not obviate the difference between the two types of cycle. This difference has implications for the criteria used to determine timing of administration of the luteinization signal before oocyte retrieval. PMID- 11939158 TI - Patient scheduling for gonadotrophin-releasing hormone antagonist protocols. AB - Gonadotrophin-releasing hormone (GnRH) antagonists offer an alternative approach to the management of superovulation in assisted conception. Although several large multicentre randomized trials have provided information concerning the safety and efficacy of GnRH antagonists, their introduction into clinical practice has identified several new problems and opportunities. In vitro fertilization (IVF) practitioners with many years of experience of the 'long protocol' of pituitary downregulation with GnRH agonists have had to manage unfamiliar problems associated with patient scheduling and IVF cycle management when introducing GnRH antagonists into practice. Antagonist cycles require greater flexibility on the part of the IVF unit, with a need for 6 (or even 7) day working patterns and altered monitoring schedules. This article addresses the possibility of using oral contraceptives as part of GnRH antagonist cycle programming, and examines the definition of a 'good' cycle when using the antagonist. PMID- 11939160 TI - The gonadotrophin-releasing hormone antagonist ganirelix--history and introductory data. AB - The gonadotrophin-releasing hormone antagonist ganirelix has recently become available to clinicians. Its indication, prevention of premature luteinizing hormone surges in assisted reproduction programmes, has been investigated extensively in numerous studies. This article summarizes the major results from pharmacokinetics studies, a double-blind dose-finding trial and three large-scale phase III randomized clinical trials, comparing ganirelix and the most commonly used gonadotrophin-releasing hormone agonists, buserelin,leuprolide and triptorelin, in a long protocol. It is concluded that controlled ovarian hyperstimulation with ganirelix offers significant advantages in terms of convenience of treatment as reflected in a considerably reduced treatment period.Safety and tolerance as well as overall clinical outcome are good. PMID- 11939159 TI - Use of gonadotrophin-releasing hormone agonists to trigger ovulation. AB - The introduction of gonadotrophin-releasing hormone (GnRH) agonists combined with gonadotrophins is considered to be one of the most significant advances in the development of in vitro fertilization (IVF) treatment. However, ovarian hyperstimulation syndrome (OHSS) remains a significant complication of controlled ovarian hyperstimulation. One possible strategy to reduce the risk of this complication would be the use of GnRH agonists instead of human chorionic gonadotrophin (hCG) to trigger the final stages of oocyte maturation. GnRH agonists are able to induce an endogenous surge of luteinizing hormone (LH) and follicle-stimulating hormone (FSH) and the effect may be more physiological than that of exogenous hCG. Several uncontrolled and controlled clinical studies have confirmed the efficacy of GnRH agonists for triggering ovulation, and pregnancy rates are comparable to those achieved with hCG. The incidence of OHSS appears to be decreased, but larger controlled studies are required to confirm this observation. The recent introduction of GnRH antagonists has led to renewed interest in the use of GnRH agonists to induce final oocyte maturation. An international multicentre randomized controlled trial has been completed recently comparing the efficacy of GnRH agonist with hCG for triggering ovulation in women undergoing controlled ovarian hyperstimulation using the GnRH antagonist ganirelix for pituitary suppression. The aim of the study was to determine the efficacy of the novel protocol for ovarian stimulation before IVF, in terms of pregnancy outcomes and the prevention of OHSS. PMID- 11939161 TI - British Society of Gastroenterology annual meeting. 17-20 March 2002. Abstracts. PMID- 11939162 TI - Abstracts from the XIV International Symposium on Drugs Affecting Lipid Metabolism. September 9-12, 2001, New York, USA. PMID- 11939163 TI - Fines in Mount Sinai death. Union officials blame layoffs for ongoing problems. PMID- 11939164 TI - Arbitration is optional. HMOs rankled by AAA's policy change. PMID- 11939165 TI - Cardiac-care hotbed. Louisiana gets four new facilities. PMID- 11939166 TI - Another heart war. Columbia, S.C., is competition battlefield. PMID- 11939167 TI - 25 largest master's programs in healthcare administration. PMID- 11939168 TI - [Outpatient diagnosis and management of generalized anxiety disorders in adults. Recommendations for clinical practice]. PMID- 11939170 TI - Cumulative author and subject index for volumes 184-194. PMID- 11939169 TI - Can't lung cancer patients detoxify procarcinogens? AB - BACKGROUND: Glutathione S-transferase mu (GST mu) enzyme detoxifies carcinogens in tobacco smoke. We assessed the clinical usefulness of serum assay of GSTm in determining the risk for lung cancer. MATERIALS AND METHODS: Fifty-nine patients with primary lung cancer and 32 control cases were enrolled. GSTm detection was performed by the method ELISA. RESULTS: GSTm enzyme positivity rate of the patient group (39%) was significantly lower than the control group (59.4%) (p < 0.05). The GSTm positivity rates were 28.6% for the non-smoker patients with a cancer history of relatives, 31.6% for the smoker patients with the cancer history of relatives, 14.6% for the non-smoker patients with the lung cancer history of relatives and 16.7% for the smoker patients with the lung cancer history of relatives. CONCLUSIONS: We concluded that if the people lacking GSTm are smokers and have a cancer and/or lung cancer history among their relatives, they would challenge a greater risk of lung cancer than the individuals having GST mu isoenzyme. PMID- 11939171 TI - A new cytotoxic stigmastane steroid from Pistia stratiotes. AB - A new cytotoxic stigmastane, 7beta-hydroxy-4,22-stigmastadien-3-one (3), together with two known cytotoxic stigmastanes (1, 2) and a norisoprenoid (4), have been isolated from the aquatic plant pistia stratiotes. The structure determination was accomplished by spectroscopic methods. PMID- 11939172 TI - Pediatric Academic Societies' annual meeting. May 4-7, 2002, Baltimore, Maryland, USA. Abstracts. PMID- 11939173 TI - High clinical inflammatory activity prior to the development of secondary progression: a prospective 5-year follow-up study. AB - OBJECTIVE: To study if there are different patterns of clinical activity- measured by the annual exacerbation rate (AER)--among relapsing-remitting multiple sderosis (RRMS), "early" secondary multiple sclerosis (SPMS) and "late" SPMS. METHODS: A prospective 5-year follow-up study in 80 MS patients has been carried out, calculating the AER and the mean expanded disability status scale (EDSS) change rate (MCR). RESULTS: A significant difference on the AER, among RRMS, early SPMS and late SPMS, has been found. CONCLUSIONS: The SPMS has a high clinical inflammatory activity before and during its transformation from a RRMS. PMID- 11939174 TI - Oral lichen planus and hepatitis C virus infection. AB - OBJECTIVE: This investigation was conducted to determine the possible association between oral lichen planus (OLP) and hepatitis C virus infection (HCV) in the population of Sao Paulo (Brazil). MATERIALS AND METHODS: Three groups of patients were studied: group 1 was composed of 68 patients with OLP lesions; group 2 had 126 patients with HCV infection; and the control group consisted of 898 individuals seeking dental treatment at our school, used to determine the prevalence of lichen planus in the general population. The prevalence of HCV in group 1 was determined and compared with that of the population of Sao Paulo (Focaccia et al (1998) Brazilian J Infect Dis 2: 269), while the prevalence of OLP in group 2 was determined and compared with that of the control group. RESULTS: The results showed that the frequency of HCV in OLP patients was 8.8%, significantly higher than in the general population of Sao Paulo, which is 1.4% (P = 0.002), and the frequency of OLP in HCV patients (4.7%) was also significantly higher (P = 0.0003) than that of the control group (0.6%). CONCLUSION: These data suggest that, at least in Sao Paulo, there is an association between OLP and HCV infection. PMID- 11939175 TI - Abstracts of the 25th German Cancer Congress. 10-14 March 2002, Berlin, Germany. PMID- 11939176 TI - Preliminary evaluation of a standard reference material for chiral stationary phases used in liquid and supercritical fluid chromatography. AB - The applicability of a new Standard Reference Material (SRM) for the evaluation of chiral stationary phase (CSP) performance was demonstrated by utilizing the SRM to characterize the chromatographic behavior of eight commercially available CSPs in liquid and supercritical fluid chromatography. The SRM consists of five ethanolic solutions, each containing one chiral compound. These test mixtures can be used to assess changes in column performance over time and to evaluate lot-to lot variability in column manufacturing. The SRM was also used to probe the effect of various parameters on column performance. PMID- 11939177 TI - An atypical interlaboratory assay: looking for an updated hallmark (-jewelry) method. AB - A comparison between a laser-induced breakdown spectrometry-partial least squares (LIBS-PLS) method and methods based on some well-known techniques, such as induced-coupled plasma-atomic emission spectrometry (ICP-AES), flame atomic absorption spectrometry (FAAS), and atomic scanning microscopy (ASM) is presented in order to both validate the content of gold and silver in alloys to be used as solid standards and develop an alternative to the established methods for the hallmark of gold and silver in jewelry pieces. 17 alloys with gold concentrations ranging from 100 to 50% and 8 alloys with silver concentrations between 100 and 80% and variable concentrations of other metals usually present in jewels were used as solid standards in LIBS in order to develop a method as general as possible. The results obtained in the analysis of some alloys (9 for gold and 7 for silver) show that the proposed method is comparable with the official one. PMID- 11939178 TI - Wavelet transform applications in analytical chemistry. AB - The wavelet transform has been established with the Fourier transform as a data processing method in analytical chemistry. The main fields of application in analytical chemistry are related to denoising, compression, variable reduction, and signal suppression. Analytical applications were selected showing prospects and limitations of the wavelet transform. An important aspect consists in showing the advantage of wavelet transform over Fourier transform with respect to dual localization of a signal in both the original and the transformed domain enabling principal new application fields in comparison with Fourier transform. PMID- 11939179 TI - Miniaturization. PMID- 11939180 TI - Evaporation rates of alkanes and alkanols from acoustically levitated drops. AB - Evaporation constants of acoustically levitated drops from the homologue series of n-alkanes and 1-alkanols in ambient air have been evaluated by size and temperature measurements. The size of the pure liquid drops, within a diameter range of 0.1 to 2.5 mm, was monitored using a CCD camera, while temperature measurements were carried out by IR thermography. During drop evaporation, water from a humid environment with a relative humidity between 5 and 80% was condensed on the drop surface and in the case of n-pentane, the condensed water froze as a result of the evaporative cooling. PMID- 11939181 TI - Mixed-gas inductively coupled plasma atomic emission spectrometry using a direct injection high efficiency nebulizer. AB - Experimental studies and computer simulations were conducted to identify plasma operating conditions and to explore and contrast the excitation conditions of Ar, Ar-O2, and Ar-He inductively coupled plasmas (ICPs) for the introduction of microliter volumes of sample solutions with a direct injection high efficiency nebulizer (DIHEN). The best MgII 280.270 nm/MgI 285.213 nm ratio (6.6) measured with Ar ICP atomic emission spectrometry for the DIHEN (RF power = 1500 W; nebulizer gas flow rate = 0.12 L min(-1)) was less than the ratio (8.2) acquired on the same instrument for conventional nebulization (1500 W and 0.6 L min(-1)). Addition of small amounts of O2 or He (5%) to the outer gas flow improved excitation conditions in the ICP, that is, a more robust condition (a MgII/MgI ratio of up to 8.9) could be obtained by using the DIHEN with Ar-O2 and Ar-He mixed-gas plasmas, thereby minimizing some potential spectroscopic and matrix interferences, in comparison to Ar ICPAES. PMID- 11939182 TI - Analysis of submicroliter samples using micro thermospray flame furnace atomic absorption spectrometry. AB - A recently described thermospray flame furnace atomic absorption spectrometric (TS-FF-AAS) system has been modified in order to extend the applicability of the method for the determination of elemental traces in very small sample volumes (microliter or submicroliter). As an easily available, effective thermospray vaporizer, a fused silica capillary was used and the liquid sample was transported by 1 MPa (10 bar) gas pressure delivered by a standard gas cylinder. A 0.3 microL sample volume can be analyzed with a higher power of detection than using 3 orders of magnitude larger sample volumes with conventional flame atomic absorption spectrometry. The relative standard deviations (N=12) for 0.3 microL volumes and 5 microg/mL Pb samples amount to 3.1% and 3.8% in signal height and signal area, respectively. The detection limit was found to be 69 ng/mL. Initial experiments with other elements (Cd, Hg, Tl, Zn) led to similar results. PMID- 11939183 TI - Genomics. PMID- 11939184 TI - Development of an assay for label-free high-throughput screening of thrombin inhibitors by use of reflectometric interference spectroscopy. AB - We describe the development of, and analytical conditions used for, parallel affinity assay for thrombin inhibitors adapted to the first label-free optical screening HTS detection set-up fully integrable into a screening platform. To achieve compatibility with pharmaceutical libraries, an HTS-transducer was realized by gluing the bottomless scaffolds of 96- and 384-well plastic microplates on to transducer slides. The transducer are coated with a dextran, to ensure biocompatibility and functionality, and a known thrombin inhibitor was attached covalently to it. By adapting reflectometric interference spectroscopy for simultaneous reading of the whole transducer plate we were able to detect the binding of thrombin in all the wells of the microplates on-line, in parallel, and time resolved. By using an inhibition assay, the screening of 384 substances for thrombin activity can be performed within an assay time of less than 15 min. We also show that the data quality is high enough for parallel quantification of the IC 50 values of the library substances. PMID- 11939185 TI - Insulin receptor binding to erythrocytes during normal pregnancy: an update of the method. AB - Normal values of the parameters of insulin receptors in erythrocytes were provided to make a control group throughout gestation, 24 h postpartum and six weeks after delivery with the aim of comparing them with other parameters with insulin receptor-related pathologies. Thus, one of the purposes of this study was the update of a method to calculate the parameters of insulin receptors in erythrocytes, carrying out several modifications that improved the assay: during binding studies incubation was in continuous rotation shaking, and increasing maximal insulin binding. Other modifications were the increase in the concentration of insulin 125I to 1 ng/mL, and the maximal concentration of unlabeled insulin, 100 ng/mL. Erythrocyte age was considered by the intracellular creatine content providing control values and allowing the normalization of the parameters of insulin receptor during gestation. Data obtained in this study indicated that changes at receptor binding level may be also considered to explain insulin resistance: week 28 showed maximal insulin secretion (16.70+/ 1.44 microU/mL) whereas plasma glucose concentrations remained almost constant (91.14+/-2.37 mg/100 mL) with respect to the 1st and 2nd trimester of pregnancy (89.73+/-1.38 and 91.71+/-2.10 mg/100 mL respectively); insulin reached the point of maximal resistance, which is explained by a decrease of maximum specific insulin binding, %Bo (6.32+/-0.51) at minimal values due to a decrease of high affinity receptor number per erythrocyte, N-AA (16+/-2) at minimal values. Moreover, the negative correlation between progesterone (31.2+/-0.2 ng/mL) and Ka AA (r=-0.71) could possibly be related to this maximal resistance. PMID- 11939186 TI - Characterization and quantification of metallothionein isoforms by capillary electrophoresis-inductively coupled plasma-isotope-dilution mass spectrometry. AB - In a new approach to the characterization and quantification of metallothionein isoforms an on-line isotope-dilution method in combination with the coupling of capillary electrophoresis (CE) to an inductively coupled plasma-sector field mass spectrometer (ICP-SFMS) is reported. Metallothionein (MT) isoforms are separated by CE and the elements Cu, Zn, Cd, and S are detected simultaneously by use of ICP-SFMS in the medium resolution mode. On-line isotope dilution is performed by continuous introduction of an isotopically enriched, species-unspecific spike solution after the separation step. MT from rabbit liver and a further purified MT-1 isoform were quantified by determination of sulfur, and the stoichiometric compositions of the metalloprotein complexes are characterized by determination of their sulfur-to-metal ratios. PMID- 11939187 TI - Sensor arrays and array sensors. PMID- 11939188 TI - Miniaturized fiber-in-tube solid-phase extraction as the sample preconcentration method for microcolumn liquid-phase separations. AB - Miniaturized fiber-in-tube solid-phase extraction (fiber-in-tube SPE) has been developed as a solventless sample preconcentration technique for microcolumn liquid-phase separation methods. Short capillaries packed with polymer filaments were employed as the extraction tube and the preconcentration power for phthalates in aqueous solutions was studied. On the basis of the successful on line coupling of this preconcentration method with liquid chromatography (LC), a more miniaturized extraction cartridge, which is installed in the rotor of the micro-injector, has been developed. With a modified commercially available valve, on-line coupling of this sample preconcentration method to capillary electrochromatography (CEC) was also investigated. PMID- 11939189 TI - Evaluation of the possibility of detecting benzenic pollutants by direct spectrophotometry on PDMS solid absorbent. AB - Solid-phase extraction has become one of the most commonly used techniques for preconcentration of analytes from environmental samples. In the standard use of solid sorbent phases the extracted pollutants are subsequently eluted with a suitable organic solvent before chromatographic analysis. An alternative to this procedure is analysis of the adsorbed and concentrated pollutants by direct application of a spectroscopic method (fluorimetry or absorptiometry) to the phase. Although this method cannot be expected to give results as precise as those given by chromatographic methods, it might have valuable applications, particularly for "on site" pollution monitoring. This paper reports an evaluation of the capability of the method for the spectrophotometric detection of BTX (benzene, toluene, xylenes) in aqueous media and in contaminated atmospheres, with polydimethylsiloxane (PDMS) as sorbent. The tests performed, with the estimated detection limits, indicate that the method is relatively simple and easy to operate and sensitive enough for application to the monitoring of pollution both in water and in air in an industrial ambient atmosphere. PMID- 11939191 TI - Matrix-assisted laser desorption ionization-mass spectrometry (MALDI-MS). PMID- 11939190 TI - A fluorescence-based sensing system for the environmental monitoring of nickel using the nickel binding protein from Escherichia coli. AB - A sensing system for nickel based on the nickel binding protein (NBP) from Escherichia coli is shown to be feasible. The versatility of NBP was demonstrated by its use in three different assay formats. When the NBP binds nickel, it undergoes a conformational change that can be used as the basis for an optical sensing system for nickel. The NBP gene was overexpressed in E. coli and the protein purified in a single step using perfusion anion-exchange chromatography. A unique cysteine residue at position 15 in the NBP was labeled with the fluorophore, N-[2-(1-maleimidyl)ethyl]-7-(diethylamino)coumarin-3-carboxamide (MDCC). In a spectrofluorimetric assay, there was a maximum of 65% quenching of the fluorescence signal produced by NBP-MDCC in the presence of nickel. A response curve for nickel using NBP-MDCC revealed a detection limit of 8 x 10(-8) mol L(-1). NBP-MDCC was also used to develop assays in microtiter plate and fiber optic bundle formats. Detection limits for nickel using these formats were also in the submicromolar range. Selectivity studies conducted with other divalent metals, including copper, cobalt, iron, cadmium, and manganese, showed that fluorescence quenching for cobalt was similar in magnitude but with a detection limit more than 10-fold higher than for nickel. The quenching responses were lower for the other metals, with detection limits at least 10 to 100 times higher than for nickel. These results suggest that fluorescently labeled NBP is potentially useful in the development of a sensing system for nickel. PMID- 11939192 TI - Capillary electrophoresis using high ionic strength background electrolytes containing zwitterionic and non-ionic surfactants and its application to direct determination of bromide and nitrate in seawater. AB - In capillary electrophoresis, it is commonly considered that even a moderately high ionic concentration in the background electrolyte (BGE) leads to high currents, resulting in Joule heating and serious peak distortion. As a new approach to overcome this problem, zwitterionic (Zwittergent-3-14) and/or non ionic (Tween 20) surfactants have been added to BGEs containing high salt concentrations (e.g. 0.3 M NaCl) and have been shown to result in acceptable separation currents (<200 microA). In turn, these BGEs could be applied to the separation of samples containing high salt concentrations (such as undiluted seawater) without the occurrence of any significant peak broadening due to electrodispersion of the sample. For example, a BGE comprising 10 mM Zwittergent 3-14, 50 mM Tween 20, 0.3 M NaCl and 5 mM phosphate (ph 7) could be used for the determination of UV-absorbing anions in seawater, giving good peak shapes and detection limits of 0.8 microM and 0.6 microM for nitrate and bromide, respectively. The beneficial effects of the non-ionic surfactant on the separation were attributed largely to suppression of the electro-osmotic flow. On the other hand, the zwitterionic surfactant was found to be capable of the incorporation of some anions in accordance with the behaviour of these same surfactants in electrostatic ion chromatography. This incorporation resulted in a decreased conductivity of the BGE and also a change in the separation selectivity of the system. PMID- 11939193 TI - Graphite furnace atomic absorption spectrometry used for determination of total, EDTA and acetic acid extractable chromium and cobalt in soils. AB - Methods for determination of both the total and extractable content of Cr and Co in soil samples were investigated. For the total content of metal, ultrasonic slurry sampling graphite furnace atomic absorption spectrometry was used and compared with conventional analyses after microwave digestion. The influence of grinding, leaching and homogeneity for slurry sampling was also examined. The concentration of the elements in the analyzed materials were in the range: 50 microg g(-1)-0.4% for Cr and 8-14 microg g(-1) for Co. Relative standard deviations (slurry sampling) were in the range 3%-12% for Cr and 0.3%-6% for Co determinations. The detection limit and characteristic mass (peak-area measurements) for Co were 0.14 microg g(-1) and 12.6 pg, respectively. For Cr less sensitive wavelengths were used. Excellent agreement with certified reference material was found for total Cr and Co using slurry sampling. EDTA and acetic acid extractions were performed, using protocols given by the Measurement and Testing Programme of the European Commission. The percentages extracted for the different soil samples were 0.3-1.0 for Cr and 2.5-24 for Co. To validate the accuracy of the extractable Cr, CRM 483 Sewage sludge amended soil was analyzed. The values found were 37% and 32% higher than the certified value for EDTA and acetic acid extractable Cr, respectively. The precision for extractable concentration of Cr and Co was about 6% or less. External calibration with aqueous standards, matched to contain the same reagents as the samples, was employed. PMID- 11939194 TI - Multi-residue analysis of polycyclic aromatic hydrocarbons, polychlorobiphenyls, and organochlorine pesticides in marine sediments. AB - A multi-residue analysis procedure using microwave-assisted extraction and pre purification has been developed for the combined analysis of polycyclic aromatic hydrocarbons (PAH), polychlorobiphenyls (PCB), and organochlorine pesticides (OCP) in marine sediments. This procedure has been validated with certified marine sediment. Several surrogate standards have been employed and the use of octachloronaphthalene (OCN) as a surrogate standard for organochlorine determination in this matrix is discussed. The recoveries of all compounds were high (>70%) and the relative standard deviations are of the same order as the certified values. Different analytical problems are discussed, including DDT degradation in gas chromatography and laboratory PCB background levels. Quantification problems encountered for two pesticides (cis-chlordane and trans nonachlor) were attributed to PAH interference in the GC-ECD chromatogram. PMID- 11939195 TI - ICP-MS. Alternative means for the elimination of interferences. PMID- 11939196 TI - A laser-induced fluorescence dual-fiber optic array detector applied to the rapid HPLC separation of polycyclic aromatic hydrocarbons. AB - A multi-channel detection system utilizing fiber optics has been developed for the laser-induced fluorescence (LIF) analysis of chromatographic eluents. It has been applied to the detection of polycyclic aromatic hydrocarbons (PAH) in a chromatographically overlapped standard mixture and to a complex soil sample extract obtained during fieldwork. The instrument utilizes dual-fiber optic arrays, one to deliver multiple excitation wavelengths (258-342 nm) generated by a Raman shifter, and the other to collect fluorescence generated by the sample at each excitation wavelength; the collected fluorescence is dispersed and detected with a spectrograph/CCD combination. The resulting data were arranged into excitation emission matrices (EEM) for visualization and data analysis. Rapid characterization of PAH mixtures was achieved under isocratic chromatographic conditions (1.5 mL min(-1) and 80% acetonitrile in water), with mid microg L(-1) detection limits, in less than 4 minutes. The ability of the instrument to identify co-eluting compounds was demonstrated by identifying and quantifying analytes in the rapid analysis of a 17 component laboratory-prepared PAH mixture and a soil extracted sample. Identification and quantification were accomplished using rank annihilation factor analysis (RAFA) using pure component standards and the EEMs of mixtures measured during the rapid high-performance liquid chromatography (HPLC) method as the unknowns. The percentage errors of the retention times (RTs) determined using RAFA compared to the known RTs measured with a standard absorbance detector were between 0 and 11%. For the standard PAH mixture, all 17 components were identified correctly and for the soil extracted sample, all 8 analytes present were correctly identified with only one false positive. Overall, the system achieved excellent qualitative performance with semi-quantitative results in the concentration predictions of both the standard mixture and the real-world sample. Electronic supplementary material to this paper can be obtained by using the Springer LINK server located at http://dx.doi.org/10.1007/s00216-001-1125-6. PMID- 11939198 TI - Chiral separations. PMID- 11939197 TI - Quantification of monohydroxy-PAH metabolites in urine by solid-phase extraction with isotope dilution-GC-MS. AB - For measurement of biomarkers from polycyclic aromatic hydrocarbon (PAH) exposure, an analytical method is described quantifying hydroxylated PAH (OH-PAH) in urine samples. This method determined monohydroxy metabolites of naphthalene, fluorene, phenanthrene, fluoranthene, pyrene, chrysene, benzo[c]phenanthrene, and benz[a]anthracene. The sample preparation consisted of enzymatic hydrolysis, solid-phase extraction and derivatization with a silylating reagent. Five carbon 13 labeled standards were used for isotope dilution. Analytes were separated by gas chromatography (GC) and quantified with high-resolution mass spectrometry (HRMS). This method produced good recoveries (41-70%), linearity, and specificity. Data were corrected for blank levels from the naphthalene, fluorene, and phenanthrene metabolites. Method detection limits ranged from 2 ng L(-1) for 1-hydroxypyrene to 43.5 ng L(-1) for 1-hydroxynaphthalene. Using quality control charts from two urine pools, the method can be readily applied to biomonitoring PAH exposure. PMID- 11939199 TI - Colourants and opacifiers in seventh and eighth century glass investigated by spectroscopic techniques. AB - Glass fragments dating from the seventh and eighth century AD were excavated in the Crypta Balbi in Rome. They were studied to detect agents involved in colour development and opacification. Reflectance spectra recorded on powdered samples revealed the contribution of Fe(II), Fe(III), Mn(III), Cu(II), and Co(II) ions in determining colour hues. The effect of the Mn/Fe atomic ratio on glass colour is discussed. It is apparent that medieval glassmakers in Italy could obtain a wide range of colours by exploiting the presence of iron and manganese as contaminants of sand and flux and controlling the amount of oxygen let into the furnace. X-ray diffraction and scanning electron microscopy coupled with energy-dispersive X-ray analysis were used to study opaque fragments. The presence of calcium antimonate was detected in white, blue, and blue-green fragments, and elemental copper was detected in a red glass. PMID- 11939200 TI - Spectrochemical analysis with lasers. PMID- 11939201 TI - The role of modern biochemical methods in the determination of hygienically relevant microorganisms in foodstuffs and (drinking) water. PMID- 11939203 TI - Detection techniques for liquid chromatography. PMID- 11939202 TI - Hyphenated techniques. PMID- 11939204 TI - Molecular electrochemistry. PMID- 11939205 TI - Laser-ablation inductively-coupled plasma mass spectrometry. PMID- 11939206 TI - Elemental speciation. PMID- 11939207 TI - Microscale liquid phase separations. PMID- 11939208 TI - Molecularly imprinted polymers. PMID- 11939209 TI - Extraction techniques. PMID- 11939210 TI - DNA chip technology. PMID- 11939211 TI - Detection trends in high throughput screening. PMID- 11939212 TI - Aptasensors--the future of biosensing? AB - Aptamers are artificial nucleic acid ligands that can be generated against amino acids, drugs, proteins and other molecules. They are isolated from combinatorial libraries of synthetic nucleic acid by an iterative process of adsorption, recovery and reamplification. Aptamers, first reported in 1990, are attracting interest in the areas of therapeutics and diagnostics and offer themselves as ideal candidates for use as biocomponents in biosensors (aptasensors), possessing many advantages over state of the art affinity sensors. The properties of aptamers, their applicability to biosensor technology, current research and future prospects are addressed in this short review. PMID- 11939213 TI - MEMS-based sample preparation for molecular diagnostics. AB - Completion of the Human Genome Project is driving the rapid development of molecular diagnostics in the laboratory. To accelerate the penetration of genetic tests and other nucleic acid-based tests into clinical markets, simple, compact, automatic sample-preparation systems for molecular diagnostics must be developed. Microelectromechanical systems (MEMS) is a promising approach for the development of automated sample preparation for the clinical laboratory or point-of-care setting. This review discusses MEMS-based components that could be applied to the different stages of the sample-preparation process such as cell separation, nucleic acid purification, and nucleic acid amplification. Examples of functional component integration are given. Issues discussed include partitioning of functions between the instrument and disposable unit, methods of propulsion of fluids and particles, vapor and liquid barriers, and sample size. Although further evaluation and development are needed to provide practical solutions to some of these issues, we conclude that MEMS-based components might contribute to some components in a sample-preparation system consisting of modular instruments and disposable units, but will not provide a generic or a totally integrated solution. PMID- 11939214 TI - Developments in ion mobility spectrometry-mass spectrometry. AB - Ion mobility spectrometry (IMS) has been used for over 30 years as a sensitive detector of organic compounds. The following is a brief review of IMS and its principles with an emphasis on its usage when coupled to mass spectrometry. Since its inception, IMS has been interfaced with quadrupole, time-of-flight, and Fourier-transform ion cyclotron resonance mass spectrometry. These hybrid instruments have been employed for the analysis of a variety of target analytes, including biomolecules, explosives, chemical warfare degradation products, and illicit drugs. PMID- 11939215 TI - Speciation of mercury, tin, and lead compounds by gas chromatography with microwave-induced plasma and atomic-emission detection (GC-MIP-AED). AB - Because of their high toxicity and widespread distribution, the reliable selective quantification of alkyl and aryl species containing mercury, tin, or lead has been one of the goals of speciation analysis in recent years. Since becoming commercially available, GC-MIP-AED has been one of the most-used tools in this work. In this paper, the value and limitations of GC-MIP-AED for the speciation of Hg, Sn, and Pb compounds in environmental samples are reviewed and compared with the analytical characteristics of other hyphenated GC-based techniques. Because quantification of Hg, Sn, and Pb species by GC techniques normally requires complex sample preparation involving several steps, the effect of sample-preparation methods on the accuracy and precision of the results is discussed. Finally, we describe the current status of a rapid, low-cost GC-MIP PED system specifically designed for routine quantification of Hg, Sn, and Pb species in environmental control laboratories. PMID- 11939216 TI - Bioentrepreneurship 2002: at home in your lab. PMID- 11939217 TI - Comparison of reflectometric interference spectroscopy with other instruments for label-free optical detection. AB - On the basis of kinetic measurements of biomolecular interactions, a reflectometric interference spectroscopy (RIfS) setup is compared with two commercial instruments. These instruments are based on evanescent wave techniques, surface plasmon resonance (SPR) (represented by BIAcore 2000) and resonant mirror (RM) technique (using IAsys plus). All methods allow a label-free and time-resolved optical detection of biomolecular interaction. These methods are mainly used in biomolecular interaction analysis (BIA). They provide practical techniques for quantifying equilibrium constants and rate constants over several orders of magnitude. The general parameters of the three detectors, namely baseline noise and drift as well as overall sensitivity and limits of detection were compared. The fluid handling and the related implications on the measurements have also been considered. The interaction between thrombine and thrombine inhibitor (TI) was investigated as a test system with the three different methods and the kinetic rate constants were determined and compared. For this TI was immobilized on the surface and binding of thrombine was monitored time-resolved. Determination of the kinetic rate constants could prove that the RIfS set-up is comparable with SPR using BIAcore 2000 and RM technique represented by IAsys plus. PMID- 11939218 TI - [Imaging of lipoma and liposarcoma]. AB - The aim of this paper is to describe the two most frequent lipomatous soft tissue tumors, lipoma and liposarcoma, and to highlight the radiologic features allowing their differentiation. PMID- 11939219 TI - [Work-up of soft tissue masses]. AB - Imaging plays a major role in the diagnosis and follow-up of soft tissue masses. Echography including color-coded and Doppler studies is performed as a first step procedure. MRI contributes essential topographic and semeiologic information but may have a poor sensitivity and specificity for the characterization of malignant lesions showing features atypical for benignity. Early recognition of malignancy enables appropriate diagnostic work-up, including biopsy, and optimal management. PMID- 11939220 TI - [Atypical ganglion cysts]. AB - Intra- or para-articular ganglion cysts are frequent and can develop at some distance from the joints. The authors describe examples of typical and atypical ganglion cysts, namely meniscal cyst, intra-articular cyst, intraosseous cyst, intraneural cyst of tibial nerve, adventitial cyst of popliteal artery, and para articular cyst of the hip, filled with gas of presumed articular origin. The diagnosis of a ganglion cyst can be difficult when there is no evident articular communication. If demonstration of communication is necessary (in case of uncertain diagnosis or for preoperative assessment), the best procedure is arthrography followed by a CT scan 1-2 hours after the injection. PMID- 11939222 TI - Sylvester O'Halloran Surgical Scientific Meeting. Limerick, Ireland, 8-9 March 2002. Abstracts. PMID- 11939221 TI - [Uterine fibroids embolization: a review]. AB - Uterine artery embolization for symptomatic leiomyomas is a new attractive treatment in patients who do not desire pregnancy and for whom conventional therapy has failed. Uterine fibroid embolization can also be considered for patients who desire pregnancy when myomectomy is technically difficult or/and in case of recurrence after myomectomy. 90% improvements are commonly reported in abnormal bleeding, pelvic pain, and in bulk-related symptoms. This technique allows reduction of the hospital stay, the convalescence period, the morbidity and the mortality rate compared to conventional surgical treatment. PMID- 11939223 TI - Recent developments in health insurance, life insurance, and disability insurance case law. PMID- 11939224 TI - Recent developments in medicine and law. AB - This article addresses important developments in the field of medicine and law during the period of September 1999 through August 2000. Although it does not discuss every reported case or piece of legislation, it broadly surveys some of the more significant developments in the medicine and law arena. PMID- 11939225 TI - Doctors, HMOs, ERISA, and the public interest after Pegram v. Hendrich. PMID- 11939226 TI - Evolving tension between HMO liability precedent and legislation. PMID- 11939227 TI - Changing conceptions of measurement validity: an update on the new standards. AB - This article serves as a follow up to a 1997 article in the Journal of Nursing Education, in which the author presented a historical overview of the ways in which views of measurement validity had changed during the past half century. The new American Educational Research Association, American Psychological Association, and National Council on Measurement in Education Standards for Educational and Psychological Testing, released in 1999, includes a revised conceptualization of validity. The key changes, including the elimination of the old "trinity" view of validity and the operationalization of validity as five types of evidence, are described in this article, and specific ways to obtain evidence of each type are provided. The article concludes with a brief discussion of some of the major continuing issues and challenges in validity theory and practice. PMID- 11939228 TI - Applying the theory of planned behavior: nursing students' intention to seek clinical experiences using the essential clinical behavior database. AB - This study examined the antecedents and determinants predictive of whether nursing students (N = 92) intend to ask for assignments to perform nursing behaviors after using a database to record essential clinical behaviors. The results of applying the theory of planned behavior (TPB) to behavioral intention using multivariant path analysis suggested that the endogenous variables, attitude and subjective norms, had a significant effect on the intention to ask for assignments to perform nursing behaviors. In addition, it was primarily through attitudes and subjective norms that the respective antecedents or exogenous variables, behavioral beliefs and normative beliefs, affected the intention to ask for assignments to perform nursing behaviors. The lack of direct influence of perceived behavioral control on intention and the direct negative impact of control belief on intention were contrary to expectations, given the tenets of the TPB. PMID- 11939229 TI - Using educational technology to teach cultural assessment. PMID- 11939230 TI - Research-based practice: applying the standard in nursing education. PMID- 11939231 TI - Implementing a community assessment course targeting an agricultural aggregate: farm families as an at-risk group. PMID- 11939232 TI - Implementation of the essential elements of service learning in three nursing courses. AB - The students' creativity, enthusiasm, energy, and excellent planning skills made a tremendously positive impression on the community agencies that participated in this project. The students applied principles of: Epidemiology, including collecting meaningful descriptive data. Community and family intervention. Community partnerships. Leadership. Cultural competence. As students visited the homes of families of different ethnic and socioeconomic status than themselves, perhaps the most poignant outcome was the opportunity for students to experience cultural sensitivity, relativity, and accommodation-Implementing the essential elements of service learning provided the foundation for a successful service learning experience. It took the time, energy, and commitment of all involved, but the end result was an educational experience that benefited students, faculty, and the community. Service learning presents an effective, unique, and valuable means for nursing students to master the objectives of community health, leadership and management, and pediatrics courses, and to prepare to function in a health care environment that demands partnering with the community to use resources more effectively to promote public well-being. PMID- 11939233 TI - Skills practice in the emergency department for ADN students. PMID- 11939234 TI - Marketing principles for a learning-service community partnership model. PMID- 11939235 TI - Learning to teach: an introduction to "Teacher Talk: New Pedagogies for Nursing". PMID- 11939236 TI - "Pitching a lecture" and "reading the faces of students": learning lecturing and the embodied practices of teaching. PMID- 11939237 TI - QALYs, standard gambles, and the expected budget constraint. AB - This paper shows that when quality adjustment factors for use in constructing QALY indices are established through answers to standard-gamble questions or similar methods, the assumptions that are made by respondents about the financial consequences of changing probabilities of illness and death are critically important for the use of QALYs in cost-utility analysis (CUA). It qualifies Meltzer's [Journal of Health Economics 16 (1997) 33] result that the cost per quality-adjusted life-year (QALY) for life-saving medical interventions should include the future consumption of those who would otherwise not have survived, by showing that its validity depends on how the QALY index has been established. The paper also shows that, contrary to a widely held notion, allocation of a fixed health care budget through CUA does not generally result in a second-best efficient allocation. Another finding is that failure to specify carefully what respondents to standard-gamble questions are supposed to assume about the financial consequences of ill health may result in a bias against providing care to older individuals. PMID- 11939238 TI - Health insurer monopsony power: the all-or-none model. AB - The textbook model of monopsony that is typically employed in analyzing health insurer monopsony power does not adequately describe the supply decision of physicians confronting a dominant health insurer. Rather than restricting purchases in order to extract a lower price for provider services, monopsonistic insurers are able to obtain discounts from providers without experiencing a decrease in quantity when physicians are forced to operate along their "all-or none" supply curve. The welfare effects of the all-or-none model of monopsony are compared to those implied by the traditional model. PMID- 11939239 TI - The economics of family planning and underage conceptions. AB - This paper examines whether improved access to family planning services for under 16 is likely to help in achieving the aim of reducing underage conceptions. A simple model of rational choice is introduced which suggests that family planning increases rates of underage sexual activity and has an ambiguous impact on underage conception and abortion rates. The model is tested on panel data on regions within the UK using two approaches. The first test is whether the 1984 Gillick ruling had a differential impact on two groups: under 16 for whom access to family planning was restricted by the ruling and older teenagers who were not affected. Secondly, attendance by under sixteens at family planning clinics, suitably instrumented, is used as a proxy for access to family planning. With both approaches, no evidence is found that the provision of family planning reduces either underage conception or abortion rates. Socio-economic variables such as children in care rates and participation rates in post-compulsory education are found to be significant predictors of underage pregnancies. PMID- 11939240 TI - Waiting times for hospital admissions: the impact of GP fundholding. AB - Waiting times for hospital care are a significant issue in the UK National Health Service (NHS). The reforms of the health service in 1990 gave a subset of family doctors (GP fundholders) both the ability to choose the hospital where their patients were treated and the means to pay for some services. One of the key factors influencing family doctors' choice of hospital was patient waiting time. However, without cash inducements, hospitals would get no direct reward from giving shorter waiting times to a subset of patients. Using a unique dataset, we investigate whether GP fundholders were able to secure shorter waiting times for their patients, whether they were able to do so in cases where they had no financial rewards to offer hospitals, and whether the impact of fundholding spilled over into shorter waiting times for all patients. PMID- 11939241 TI - Genetic testing when there is a mix of compulsory and voluntary health insurance. AB - When the insurer has access to information about test status, genetic insurance can handle the negative effects of genetic testing on insurance coverage and income distribution. Hence, efficient testing is promoted. When information about prevention and test status is private, two types of social inefficiencies may occur; genetic testing may not be done when it is socially efficient and genetic testing may be done although it is socially inefficient. The first type of inefficiency is shown to be likely for consumers with compulsory insurance only, while the second type of inefficiency is more likely for those who have supplemented the compulsory insurance with substantial voluntary insurance. This second type of inefficiency is more important the less effective prevention is. It is therefore a puzzle that many countries have imposed strict regulation on the genetic information insurers have access to. A reason may be that genetic insurance is not yet a political issue, and the advantage of shared genetic information is therefore not transparent. PMID- 11939242 TI - The estimation of a preference-based measure of health from the SF-36. AB - This paper reports on the findings of a study to derive a preference-based measure of health from the SF-36 for use in economic evaluation. The SF-36 was revised into a six-dimensional health state classification called the SF-6D. A sample of 249 states defined by the SF-6D have been valued by a representative sample of 611 members of the UK general population, using standard gamble. Models are estimated for predicting health state valuations for all 18,000 states defined by the SF-6D. The econometric modelling had to cope with the hierarchical nature of the data and its skewed distribution. The recommended models have produced significant coefficients for levels of the SF-6D, which are robust across model specification. However, there are concerns with some inconsistent estimates and over prediction of the value of the poorest health states. These problems must be weighed against the rich descriptive ability of the SF-6D, and the potential application of these models to existing and future SF-36 data set. PMID- 11939243 TI - Asymmetric information, ownership and quality of care: an empirical analysis of nursing homes. AB - Theoretically, when asymmetric information exists, nonprofit organizations, due to the attenuation of the property right, provide better quality of service than do the for-profits. Despite extensive theoretical examination of the behavior of nonprofits, there has been very little empirical testing of the plausibility of these theories. This article addresses the effect of ownership type on the quality of service in the nursing home industry, an environment particularly conducive to identifying the existence of asymmetric information. The study shows that the differences between for-profit and nonprofit homes do become manifest when asymmetric information is present. PMID- 11939244 TI - Medicaid expansions and welfare contractions: offsetting effects on prenatal care and infant health? AB - Most states have adopted administrative measures to encourage the use of prenatal care among medicaid-eligible women. At the same time, declining welfare caseloads have caused many women to lose medicaid. We examine the effects of changes in income eligibility, administrative procedures, and welfare caseloads using data from all birth certificates for 1990-1996. Higher income cutoffs increased use of prenatal care, while decreases in welfare caseloads reduced it. Changes in income cutoffs also reduced fetal deaths. These results suggest that the administrative reforms have not broken the close link between welfare participation and access to medicaid. PMID- 11939245 TI - How do doctors behave when some (but not all) of their patients are in managed care? AB - Most physicians today treat a variety of patients within their practices where a variety of insurance arrangements co-exist. In this paper, we propose several theoretical explanations for physician treatment patterns when the patient population is heterogeneous at the practice level. Data from the 1993-1996 National Ambulatory Medical Care Survey (NAMCS) are used to test how practice level managed care penetration affects treatment intensity. Practice composition has strong effects on treatment. Visit duration appears to be constant across patients within a practice, while medications prescribed appear to be converging as managed care penetration increases. PMID- 11939246 TI - Where do interventions come from? PMID- 11939247 TI - Chronic illness experience: insights from a metastudy. AB - Concurrent with the recent enthusiasm for qualitative research in the health fields, an energetic call for methods by which to synthesize the knowledge has been generated on various substantive topics. Although there is an emerging literature on meta-analysis and metasynthesis, many authors overestimate the simplicity of such approaches and erroneously assume that useful knowledge can be synthesized from limited collections of study reports without a thorough analysis of their theoretical, methodological, and contextual foundations and features. In this article, the authors report some of the insights obtained from an extensive and exhaustive metastudy of qualitative studies of chronic illness experience. Their findings reveal the complexities inherent not only in any phenomenon of interest to health researchers but also in the study of how we have come to know what we think we know about it. PMID- 11939248 TI - Postpartum depression: a metasynthesis. AB - Postpartum depression has been described as a dangerous thief that robs mothers of the love and happiness they expected to feel toward their newborn babies. Even though the number of qualitative studies on postpartum depression is increasing, knowledge development will be impeded unless the rich understandings gleaned from these studies are synthesized. Using Noblit and Hare's 1988 approach, the author conducted a metasynthesis of 18 qualitative studies on postpartum depression. Four overarching themes emerged that reflected four perspectives involved in postpartum depression: (a) incongruity between expectations and the reality of motherhood, (b) spiraling downward, (c) pervasive loss, and (d) making gains. Implications for clinical practice and theory development are addressed. PMID- 11939249 TI - Trust and transparency in human encounters in tuberculosis control: lessons learned from Vietnam. AB - The purpose of this study was to elucidate understanding of mechanisms of the tuberculosis sickness trajectory in Vietnam. In-depth interviews with 24 TB patients and 16 health care providers involved in TB care were concurrently conducted, coded, and analyzed according to the grounded theory method. A need for quality interaction based on transparency and trust among patient, family, society, and provider was observed. Providers lacked an awareness of differences between male and female patients with regard to social situation and support. In particular, female patients needed social support, as they received less support from their families than men did. A need for active intervention to decrease social isolation and increase social support was more pronounced in families with a female TB patient. PMID- 11939250 TI - Active consideration: conceptualizing patient-provided support for spouse caregivers in the context of prostate cancer. AB - In this study, the authors examine the under-investigated topic of patient provided support for spouse caregivers. Thirty-four men with prostate cancer and their female partners were interviewed separately three times: before the man's radical prostatectomy, 8 to 10 weeks postsurgery, and 1 year postsurgery. The core category of active consideration encompassed 4 dimensions: easing spousal burden, keeping us up, maintaining connection, and considering spouse. Patient provided support entails two overlapping tasks: minimizing the practical and emotional impact of the illness and tending to the caregiver's social-emotional needs. A theory expounding on the double bind of being both a patient and an agent in light of masculine socialization practices is articulated and brought to bear on the phenomenon of patient-provided support. PMID- 11939251 TI - A metasynthesis: mothering other-than-normal children. AB - The author used Noblit and Hare's 1988 comparative method of synthesizing qualitative studies to address the need for collective knowledge development related to mothering other-than-normal children. Twelve studies were included in a metasynthesis for a total sample of 79. The nature of the child's disability, demographics, and methodology used varied widely. Initially, 13 common themes were extracted using reciprocal translation. Further analysis revealed 4 steps common to the mothering experience under which themes were categorized. Suggestions for application to practice include keeping in mind qualities of a supportive health professional from the mothers' perspective, encouraging mothers to challenge societal definitions of normalcy, and recognizing the significance of hope in fueling maternal caregiving. PMID- 11939252 TI - "Outing" the researcher: the provenance, process, and practice of reflexivity. AB - To increase the integrity and trustworthiness of qualitative research, researchers need to evaluate how intersubjective elements influence data collection and analysis. Reflexivity--where researchers engage in explicit, self aware analysis of their own role--offers one tool for such evaluation. The process of engaging in reflexive analysis, however, is difficult, and its subjective, ambiguous nature is contested. In the face of challenges, researchers might retreat from engaging in the process. In this article, the author seeks to "out" the researcher's presence by exploring the theory and practice of reflexivity. Examples from research illustrate its problematic potential. PMID- 11939253 TI - Doing a doctorate using a participatory action research framework in the context of community health. AB - The use of participatory action research (PAR) for doctoral research in a health setting is still a fairly rare phenomenon. The author explores what is meant by PAR and examines some of the obstacles to undertaking PAR for doctoral research so that these constraints can be overcome. The barriers are seen in terms of methodological conflicting interests, timing, and the information generated. There are also institutional obstacles. The author expresses the viewpoint that it is essential for academic institutions to consider how to minimize such obstacles for students to be able to engage in PAR in a meaningful way. PMID- 11939254 TI - Conducting research with visually impaired older adults. AB - Due to the frequency of eye disorders among older adults, qualitative researchers who involve older individuals in their work must be sensitive to the multiple ways in which visual deficits can influence the research process. The author addresses some of the difficulties encountered, insights gained, and strategies developed while conducting a phenomenological study in which all the participants were severely visually impaired. The author's insights, drawn from personal experiences, reflections, and log entries kept throughout the study, are shared to help other researchers design and implement studies in which the voices of individuals with severe visual impairments can be skillfully tapped. PMID- 11939255 TI - Seeking Emirati women's voices: the use of focus groups with an Arab population. AB - Focus groups have gained increasing acceptability as a data collection technique in qualitative research in recent years. Although used extensively with Western populations, they have been used only in a limited way in cross-cultural research. The authors describe a series of focus groups with women of childbearing age in the United Arab Emirates. The purpose was to identify Emirati women's health needs as a prelude to planning additional services and programs. Recommendations are outlined, with the appropriate cultural modifications for using this data collection tool. Major considerations that might be relevant to other parts of the Arab world include timing, location, topic, group composition, culture, religion, and selection of a facilitator and translator. PMID- 11939256 TI - Relationship between aplastic anemia and paroxysmal nocturnal hemoglobinuria. AB - Since aplastic anemia-paroxysmal nocturnal hemoglobinuria syndrome was reported in 1967, the overlap of idiopathic aplastic anemia (AA) and paroxysmal nocturnal hemoglobinuria (PNH) has been well known. The link between the 2 diseases became even more evident when immunosuppressive therapy improved survival of patients with severe AA. More than 10% of patients with AA develop clinically evident PNH. Moreover, flow cytometric analysis demonstrates that the majority of patients with AA have a subclinical percentage of granulocytes with PNH phenotype. Some of them have clearly recognizable PNH clones. Granulocytes with a PNH phenotype are also often found in normal individuals, though at much smaller percentages of cells. This finding suggests that a PNH clone is expanded in AA. consistent with a hypothesis that blood cells from patients with PNH are more resistant to an autoimmune environment. Survival of PNH clones in pathologic bone marrow may account for limited expansion of PNH clones; however, additional genetic change(s) that confers cells with growth phenotype may be required for the full development of PNH. PMID- 11939257 TI - Molecular pathogenesis of fanconi anemia. AB - Fanconi anemia (FA) is a rare autosomal recessive chromosomal breakage disorder characterized by the childhood onset of aplastic anemia, developmental defects, cancer susceptibility, and cellular hypersensitivity to DNA-cross-linking agents. FA patients can be divided into at least 8 complementation groups (FA-A, FA-B, FA C, FA-D1, FA-D2, FA-E, FA-F, and FA-G). FA proteins encoded by 6 cloned FA genes (FANCA, FANCC, FANCD2, FANCE, FANCF, and FANCG) cooperate in a common pathway, culminating in the monoubiquitination of FANCD2 protein and colocalization of FANCD2 and BRCA1 proteins in nuclear foci. These BRCA1 foci have been implicated in the process of homologous recombination-mediated DNA repair. In this review, we will summarize the current progress in the field of FA research and highlight some of the potential functions of the FA pathway in DNA-damage response. PMID- 11939259 TI - Stem cell transplantation for aplastic anemia. AB - Survival of patients with aplastic anemia treated with transplantation of bone marrow has improved significantly over the past several decades. Allogeneic bone marrow transplantation (BMT) for patients with HLA-identical siblings is now the first-line therapy, and long-term survival of approximately 90% can be expected with cyclophosphamide/antithymocyte globulin conditioning and postgrafting methotrexate/cyclosporine immunosuppression. The outcome of unrelated donor BMT has also improved significantly with the identification of a preparative regimen with less toxicity combined with the development of high-resolution DNA-based HLA typing to identify the optimal unrelated marrow donor. Patients with fully HLA matched unrelated donors should be considered candidates for transplantation prior to exposure to repeat courses of immunosuppression. Future progress in hematopoietic stem cell transplantation for aplastic anemia will be directed toward further decreasing the acute toxicity and decreasing the delayed effects of the conditioning regimens while maintaining highly reliable rates of sustained engraftment with prevention of acute and chronic graft-versus-host disease. PMID- 11939260 TI - Allele frequencies and molecular genotyping of the ABO blood group system in a Kuwaiti population. AB - The phenotypic distributions of observed numbers of ABO blood groups in a Kuwaiti sample population of 18,558 subjects are 4962 (26.7%) with A, 4,462 (24.1%) with B, 858 (4.6%) with AB, and 8,276 (44.6%) with 0. The calculated gene frequencies are 0.6678 for ABO*O, 0.1768 for ABO*A, and 0.1554 for ABO*B. Molecular genotyping of the ABO blood group system in a Kuwaiti sample population was determined using polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) analysis. The positions of nucleotides 258 and 700 of cDNA from A transferase were amplified by PCR. The amplified DNA was subjected to RFLP analysis to distinguish A, B, and O alleles. Blood samples of known ABO phenotype from 101 healthy unrelated Kuwaiti individuals (A, 29; B, 23; AB, 14; O,35) were used. Two DNA fragments of the ABO locus were designed to be amplified by 2 pairs of primers. To identify the 258th nucleotide, a 199- or 200-bp DNA fragment was amplified by PCR and digested with KpnI. For the 700th nucleotide, a 128-bp DNA fragment was amplified by PCR and digested with AluI. By analyzing the electrophoresis patterns,ABO genotypes were conclusively determined by examining the DNA fragments. The ABO genotypes of the known 101 samples were as follows: AA, 4.30%; AO, 24.41%; BB, 4.16%; BO, 24.2%; AB, 8.46%; and 00, 34.65%. These results were confirmed statistically using the calculated frequencies of IA, IB, and IO alleles. PMID- 11939258 TI - Immunosuppressive treatment of acquired aplastic anemia and immune-mediated bone marrow failure syndromes. AB - Modern therapeutic strategies for the treatment of acquired aplastic anemia are based on the current understanding of its pathophysiology as well as empiric observations. Most cases of aplastic anemia appear to be the result of immune mediated destruction of hematopoietic cells, which can be approached by stem cell transplantation in younger patients with appropriate histocompatible donors or by immunosuppression to reduce T-cell activity. Popular treatment regimens combine antithymocyte globulin with cyclosporine. Although a majority of patients respond with improved blood counts and achieve transfusion-independence, late clonal complications of myelodysplasia and cytogenetic abnormalities occur in a substantial minority of cases. Additionally, there is no clear algorithm for the treatment of refractory disease. Newer methods of treatment, including high-dose cyclophosphamide and the development of potentially tolerizing combinations of drugs. are under study. Effective therapies for aplastic anemia might also be applied to other T-cell mediated, organ-specific human diseases. PMID- 11939261 TI - Comparison of Pgp- and MRP-mediated multidrug resistance in leukemia cell lines. AB - Drug resistance is a major cause of the failure of anticancer chemotherapy. Multidrug resistance is often caused by overexpression of the P-glycoprotein (Pgp) or the multidrug resistance-related protein (MRP). In the present study, we compared daunorubicin (DNR) accumulation, subcellular distribution, and the effect of modulators on drug accumulation and subcellular distribution in the Pgp expressing K562 cell line and the MRP-expressing HL60 cell line using reverse transcriptase polymerase chain reaction, MTT (3-[4, 5-dimethylthiazol-z-yl]-2,5 diphenyltetrazolium bromide) drug cytotoxicity assay, fluorocytometry, and confocal laser scanning microscopy. The 2 resistant cell lines exhibit similar levels of resistance to DNR and decreased drug accumulation. Altered drug subcellular distribution in the resistant cell lines compared to that in the sensitive cell lines was shown and, moreover, differences in drug distributions between the 2 resistant cell lines were found. DNR fluorescence in the resistant HL60 cell line was distributed into punctate regions in the cytoplasm; the nucleus and other cytoplasm were almost negative. In contrast, the resistant K562 cells showed a bright fluorescent signal located in the peripheral cytoplasm and perinuclear region; the nucleus and other cytoplasmic regions showed no signal. Use of the modulator verapamil increased drug accumulation and restored the altered subcellular distribution of the drug in the 2 resistant cell lines. The Golgi apparatus inhibitor brefeldin A had similar action in the resistant HL60 line but had little effect in the resistant K562 line. Therefore, our study suggested that there were differences between the 2 resistant cell lines in the compartments sequestering DNR. PMID- 11939262 TI - Expression of the antiapoptosis gene survivin in human leukemia. AB - Loss of the inhibition of apoptosis is important in leukemogenesis and may influence the prognosis. Survivin is an inhibitor of apoptosis that shows selective expression during fetal development and in human malignancies. Survivin expression was examined in human leukemias using the reverse transcriptase polymerase chain reaction. Survivin gene expression was detected in 17 of 31 patients with acute myelocytic leukemia and 11 of 16 patients with acute lymphocytic leukemia but was not identified in normal bone marrow cells. Survivin expression was lower in patients with M3 acute myelocytic leukemia than in patients with other types of acute leukemia. Survivin was not detected in the chronic phase of chronic myelocytic leukemia but was observed in 5 of 7 patients with chronic myelocytic leukemia in blastic crisis. These findings suggest a relationship between survivin gene expression and hematopoietic cell differentiation. In fact, survivin gene expression was down-regulated during the differentiation of HL-60 cells after treatment with dimethyl sulfoxide or all trans-retinoic acid. Moreover, the disease-free survival rates of patients with survivin expression were lower than in patients without survivin expression. Accordingly, survivin may have a role in leukemogenesis as well as in other malignancies. Detecting survivin may also provide prognostic information. PMID- 11939263 TI - Quantitative assessment of minimal residual disease in childhood lymphoid malignancies using an allele-specific oligonucleotide real-time quantitative polymerase chain reaction. AB - We developed an assay using a real-time quantitative polymerase chain reaction (RQ-PCR) for the quantitative assessment of minimal residual disease (MRD) in childhood lymphoid malignancies by using a consensus V-region probe combining a allele-specific oligonucleotide (ASO) reverse primer. Our strategy employs a set consisting of a consensus V-region probe, an ASO reverse primer, and a patient specific forward primer for clonal antigen-receptor (IgH, immunoglobulin heavy chain; TCR, T-cell receptor) gene rearrangements (IgH-ASO and TCR-ASO RQ-PCR assays). The limit of detection in both assays was 5 copies of the target/10(5) cell equivalents. We tested the assays in 17 childhood malignancies (14 cases of acute lymphoblastic leukemia and 3 of non-Hodgkin's lymphoma). High correlation coefficients of the standard curves (>0.980) and PCR efficiency (>0.95) were achieved with all primer/probe sets. In 2 (12%) of the 17 patients, ASO primers could not be designed because there was no junctional N-sequence. The quantitative data suggest that the copy number of clonal antigen receptors markedly decreased after induction therapy in 15 of 17 patients and that 1 patient relapsed and died of the disease. Consensus probes make it possible to examine a large number of patients with only a limited number of probes. The strategy used for IgH-ASO and TCR-ASO RQ-PCR assays is accurate and reliable in the clinical prospective study of MRD in childhood lymphoid malignancies. PMID- 11939264 TI - Risk of etoposide-related acute myeloid leukemia in the treatment of Epstein-Barr virus-associated hemophagocytic lymphohistiocytosis. AB - We studied the impact of etoposide on the prognosis of 81 patients (77 of whom were children <15 years old) with Epstein-Barr virus-associated hemophagocytic lymphohistiocytosis (EBV-HLH). The study group received a median cumulative dose of 1,500 mg/m2 etoposide (range, 0-14,550 mg/m2), with a median follow-up period of 44 months (range, 20-88 months) from the diagnosis. Only 1 patient, who received 3150 mg/m2 etoposide, developed therapy-related acute myeloid leukemia (t-AML), at 31 months after diagnosis. Excluding 9 patients who underwent hemopoietic stem cell transplantation during the course of treatment, the prognosis was poorer for those patients who received less than a 1,000 mg/m2 cumulative dose of etoposide. Our results indicate that the risk of etoposide related t-AML is low. An appropriate dosage of etoposide for the treatment of EBV HLH would be in the range of 1,000 to 3,000 mg/m2. However, even at these doses, care must be taken to prevent the rare risk of t-AML. PMID- 11939265 TI - MLL gene rearrangement in acute myelogenous leukemia after exposure to tegafur/uracil. AB - We report a case of acute myelogenous leukemia (AML) with MLL (myeloid-lymphoid leukemia or mixed-lineage leukemia) gene rearrangement after exposure to tegafur/uracil. Cytogenetic and clinical findings in this patient: t(11;17) (q23;q25), AML-M4 morphology, development of AML within a short latent period after first exposure to tegafur/uracil, and good response to remission induction chemotherapy but short remission duration, have been considered typical features of therapy-related acute myelogenous leukemia (t-AML) after exposure to topoisomerase II-targeting agents. This case report suggests that t-AML may develop after exposure to tegafur/uracil and that MLL gene rearrangement may not necessarily be specific to t-AML after exposure to topoisomerase II-targeting agents. PMID- 11939267 TI - Acute myelogenous leukemia concurrent with untreated chronic lymphocytic leukemia. AB - We report a case of acute myelogenous leukemia (AML) concurrent with untreated chronic lymphocytic leukemia (CLL). An 84-year-old Japanese man was admitted to the Chihaya Hospital with persistent high-grade fever. Morphologic observation of peripheral blood and bone marrow smears revealed a proliferation of blasts and lymphocytosis with small and mature phenotypes. Immunophenotyping of the blast cells revealed CD13+, CD33+, CD34+, and HLA-DR+, and that of the lymphocytes revealed CD5+, CD19+, CD20+, and lambda+ on the cell surface. The peripheral lymphocytes revealed an IgH gene rearrangement. Chromosome analysis of 20 metaphase cells from bone marrow showed numerous abnormalities, containing +8,+11,+21. The patient's disease was diagnosed as AML with trilineage dysplasia concurrent with CLL. The simultaneous occurrence of AML and CLL is extremely rare but should not be overlooked as a possible underlying cause of lymphocyte abnormalities in AML patients. PMID- 11939266 TI - Two independent clones in myelodysplastic syndrome following treatment of acute myeloid leukemia. AB - We describe a 55-year-old Japanese woman with therapy-related myelodysplastic syndrome (t-MDS) with 2 independent clones, t(1;2)(p36;p21) and t(11;12)(pl5;ql3). She was diagnosed with acute myeloid leukemia (AML) with cytological features of the bone marrow and peripheral blood. Cytogenetic evaluation revealed a 46,XX karyotype. She received chemotherapy and achieved complete remission (CR). Despite maintenance chemotherapy, she suffered a relapse. Chromosomal analysis showed t(1;2)(p36;p21) in 2 of 20 metaphases. At second CR, this clone transiently disappeared. Nine months later, t(1;2) (p36;p21) was detected again in 3 of 20 metaphases while the patient remained in CR. Six months later, bone marrow examination disclosed trilineage dysplasia without an excess of blasts, suggesting MDS. t(1;2)(p36;p21) was observed in 16 of 20 metaphases. The clinical course and serial cytogenetic findings were diagnostic of t-MDS. The duration of t-MDS was 6 years. During this period, persistent t(1;2)(p36;p21) and transient t(11;12)(p15;q13) were found. When t-MDS evolved toAML, cytogenetic evaluation revealed 46,XX,t(1;2)(p36;p21),del(7)(q22),add(19)(p13). PMID- 11939268 TI - Bone marrow cytogenetic complete remission achieved by interferon-alpha plus cytarabine ocfosfate therapy in a patient with chronic myelogenous leukemia during extramedullary blast crisis. AB - We report a case of chronic myelogenous leukemia (CML) in which the bone marrow achieved cytogenetic complete remission (CCR) achieved by treatment with interferon-alpha and oral cytarabine ocfosfate after extramedullary blast crisis. A 51-year-old Japanese man diagnosed with CML was treated with interferon-alpha. Two months later; lymph node swellings developed in his neck and inguinal regions. Lymph node biopsy revealed the infiltration of blast cells showing bcr abl fusion signal by fluorescence in situ hybridization analysis. Bone marrow aspiration and cytogenetic analysis demonstrated that his bone marrow was still in the chronic phase, with minor cytogenetic response. Continuing interferon alpha for 6 months in combination with oral cytarabine ocfosfate resulted in the disappearance of the neck lymph node swellings followed by CCR in the bone marrow. However, rapid reenlargement of the neck and inguinal lymph nodes was noted 2 months after CCR despite maintaining medullary remission with major cytogenetic response. Finally, medullary crisis was noted 13 months from the initial development of the extramedullary crisis. This case suggests that interferon-alpha plus cytarabine ocfosfate therapy may be of benefit in the treatment of extramedullary blast crisis of CML. PMID- 11939269 TI - Successful treatment of nasal T-cell lymphoma with a combination of local irradiation and high-dose chemotherapy. AB - Nasal natural killer (NK)/T-cell lymphoma is characterized by an aggressive clinical course and poor prognosis. The term "NK/T-cell" lymphoma includes both the NK-cell type and the T-cell type, which are classified by immunophenotyping and according to T-cell receptor (TCR) rearrangement. In addition, CD56+ T-cell lymphoma is defined as NK-like T-cell lymphoma. This report concerns a 54-year old woman with nasal T-cell lymphoma. Its phenotype showed pure T-cell type with CD3+, CD56-, and TCR+ accompanied by Epstein-Barr virus infection. Although the lesions were localized in the nasal mucosa and facial skin (stage IE), local irradiation could not achieve complete remission (CR). We then administered 5 courses of CHOP (cyclophosphamide, doxorubicin, vincristine, and prednisolone) regimen followed by high-dose chemotherapy with an autologous peripheral blood stem cell transplantation. This therapy resulted in CR. Our results suggest that this lymphoma subtype may be cured by means of intensive treatment soon after diagnosis. PMID- 11939270 TI - Treatment of a child with myeloid/NK cell precursor acute leukemia with L asparaginase and unrelated cord blood transplantation. AB - A 2-year-old Japanese boy who presented with multiple cervical, axillary, and inguinal lymphadenopathy was diagnosed by immunocytochemical analysis as having myeloid/natural killer (NK) cell precursor acute leukemia. Leukemic blasts in the bone marrow were positive for CD56 (NK marker), CD7 (T-cell marker), CD33 (myeloid marker), CD34, and HLA-DR. Tumor cells in a lymph node were also positive for CD2, cytoplasmic CD3 (T-cell marker), CD7, CD33, CD34, and CD56, but negative for peroxidase staining and other T-cell, NK, and myeloid markers. Southern blot analysis showed no rearrangement bands for T-cell receptor delta and immunoglobulin heavy chain. Chromosomal analysis revealed 46,XY,inv(7)(p21q21). Neither chemotherapy for acute lymphoblastic leukemia nor that for acute myeloid leukemia induced remission in this patient. However, complete remission was achieved by the administration of L-asparaginase (6,000 U/m2 for 5 days). Because the disease was considered refractory to standard chemotherapy, cord blood transplantation was performed from an HLA 1-locus mis matched unrelated donor. The conditioning regimen consisted of total body irradiation, cytarabine, and cyclophosphamide, and cyclosporine and short-term methotrexate were employed for graft-versus-host disease (GVHD) prophylaxis. Hematological reconstitution was rapid, and only grade I acute GVHD was observed. The patient has been in remission for more than 24 months after transplantation. Our findings indicate that combination therapy with L-asparaginase and allogeneic stem cell transplantation may be useful for the treatment of myeloid/NK cell precursor acute leukemia. PMID- 11939271 TI - A perspective on the oxidative burst in the phagosome of the leukocyte and its neoplastic transformation. PMID- 11939272 TI - Sixth International Symposium on Myelodysplastic Syndromes, Stockholm, June 14 17, 2001. PMID- 11939273 TI - The Fourth Nagoya International Blood and Marrow Transplantation Symposium: new horizons in allogeneic hematopoietic cell transplantation--2001 revolution. AB - In this symposium, we saw new horizons in allogeneic transplantation. Are these truly revolutionary? We do not yet know the answer. However, there is no question about the importance of allogeneic T cells. T cells are much more powerful than any pharmacological drug man has ever generated. The question is, how do we take the most advantage of their potential. Every participant was encouraged to search for good answers to this question until the next meeting. PMID- 11939274 TI - A seriously playful man: Ernest "Jack" Hilgard's exploration of the unusual. PMID- 11939275 TI - How practitioners (and others) can make scientifically viable contributions to clinical-outcome research using the single-case time-series design. AB - Although clinicians typically possess considerable interest in research, especially about which interventions do and do not work, all too often they dismiss the notion that they themselves can make viable scientific contributions to the outcome literature. This derives from an unfortunate assumption that the only true experiment is a between-groups experiment. There is another form of true experiment that is perfectly compatible with real-world clinical practice: the single-case time-series design. Intensive and systematic tracking of one or a few patients over time can yield viable inferences about efficacy, effectiveness, and, under some circumstances, mechanism of change. This paper describes how clinicians working with hypnosis can carry out such research. The rationale and essential features of time-series studies are outlined; each design is illustrated with actual studies from the hypnosis literature; and new methods of statistical analysis, well within the statistical competence of practitioners, are described. PMID- 11939276 TI - The efficacy of hypnosis in the treatment of pruritus in people with HIV/AIDS: a time-series analysis. AB - Pruritus, or generalized itch, is a source of serious discomfort and distress in a significant minority of people living with AIDS. Anecdotal reports suggest hypnosis might be a useful treatment, leading to reductions in distress and improvements in the condition. But empirical examination of the question is notably lacking. This time-series study reports results of a 6-session self hypnosis treatment (relaxation, deepening, imagery, and home practice) for 3 HIV positive men suffering from pruritus, related to disease progression and/or HIV medications. Posttreatment, all 3 patients reported significant reductions in daily itch severity and extent of sleep disturbance due to itch. One patient also evidenced significantly less itch distress. Another also experienced significantly less time bothered by itch. For the 2 patients on which 4-month follow-up data were available, treatment benefit across variables was stable or further improved. PMID- 11939277 TI - Hypnosis for the control of HIV/AIDS-related pain. AB - This intensive case study used an A-B time-series analysis design to examine whether 5 adult patients with various AIDS-related pain symptoms benefited from a hypnosis-based pain management approach. The 3 dependent variables in this study were: (a) self-ratings of the severity of pain, (b) self-ratings of the percentage of time spent in pain, and (c) amount of p.r.n. pain medication taken. Data were collected over a period of 12 weeks, including a 1-week baseline period and an 11-week treatment period. Autoregressive integrated moving-average (ARIMA) models were used to determine the effects of the hypnotic intervention over and above autoregressive components in the data. All 5 patients showed significant improvement on at least 1 of the 3 dependent variables as a result of the hypnotic intervention. Four of the 5 patients reported using significantly less pain medication during the treatment phase. PMID- 11939278 TI - Case study examining the efficacy of a multi-modal psychotherapeutic intervention for hypertension. AB - This study examines the effectiveness of a multimodal psychotherapeutic approach using hypnosis in the treatment of a single case of hypertension. A systematic eclectic conceptualization and treatment approach was implemented using psychodynamic, behavioral, and cognitive-behavioral elements. Hypnosis was used to support each of the treatment modalities. Time-series analysis procedures indicate that the psychological interventions were associated with significantly reduced diastolic blood pressure. Additionally, the effect of the psychological interventions was significant over and above traditional pharmacological interventions. However, psychotherapeutic interventions had no substantial impact on systolic pressure. The flexibility of hypnosis as a therapeutic tool is discussed in terms of potential advantages in treatment. PMID- 11939279 TI - Salient findings in the hypnosis literature: April 2002. AB - Four important investigations were reported during the latter part of 2001. All address the biological impact of hypnotic interventions. Three of these studies focus specifically on if and how hypnotic interventions affect immune functions. A range of immune assays is employed, from allergic response to blood-based assays of immune functioning during nonlaboratory periods of stress. In all 3 cases, measurable shifts in immune functioning are associated with hypnotic interventions. A 4th compares the pattern of event-related brain potentials (ERPs) associated with hypnotic analgesia interventions and standard distraction protocols during exposure to pain. PMID- 11939280 TI - In memoriam: Ernest Ropiequet Hilgard, 1904-2001. PMID- 11939281 TI - Atmospheric dynamics and ozone cycles related to nitrogen deposition in the western Mediterranean. AB - Recent results from EU-sponsored projects have shed new light on the workings of the atmosphere in the Mediterranean Basin, where a large airmass becomes trapped and nearly stagnant over the sea during the summer. This work reviews the developments which have led to our present understanding of air pollution dynamics and associated meteorological processes in this region. In summer, the sea breezes combine with upslope winds to create recirculations along the coasts and within the western Mediterranean basin, with residence times in the order of days. Under strong insolation these recirculations become "large natural photo chemical reactors" where most of the NOx emissions and other precursors are transformed into oxidants, acidic compounds, aerosols and ozone, which can exceed European Union directives for several months of the year. The objective of this paper is to evaluate available experimental evidence and complementary modelling results to help in the interpretation of observed ozone cycles and to anticipate possible scenarios for anthropogenic nitrogen deposition in the region. PMID- 11939282 TI - Summer-time distribution of air pollutants in Sequoia National Park, California. AB - Concentrations of air pollutants were monitored during the May November 1999 period on a network of forested sites in Sequoia National Park, California. Measurements were conducted with: (1) active monitors for nitric oxide (NO), nitrogen dioxide (NO2) and ozone (O3); (2) honeycomb denuder/filter pack systems for nitric acid vapor (HNO3), nitrous acid vapor (HNO2), ammonia (NH3), sulfur dioxide (SO2), particulate nitrate (NO3-), ammonium (NH4+), and sulfate (SO4(2 )); and (3) passive samplers for O3, HNO3 and NO2. Elevated concentrations of O3 (seasonal means 41-71 ppb), HNO3 (seasonal means 0.4-2.9 microg/m3), NH3 (seasonal means 1.6-4.5 microg/m3), NO3 (1.1-2.0 microg/m3) and NH4+ (1.0-1.9 microg/m3) were determined. Concentrations of other pollutants were low. With increasing elevation and distance from the pollution source area of O3, NH3 and HNO3 concentrations decreased. Ammonia and NH4+ were dominant N pollutants indicating strong influence of agricultural emissions on forests and other ecosystems of the Sequoia National Park. PMID- 11939283 TI - Nitrogen deposition in Mediterranean forests. AB - Atmospheric deposition of inorganic nitrogen was studied at two forested sites in the Montseny mountains (northeast Spain), peripheral to the Barcelona conurbation, and at a nearby lowland town, using bulk deposition, wet-only deposition, throughfall, and dry deposition inferred from branch-washes and surrogate surfaces (metacrylate plates). Bulk deposition inputs of ammonium and nitrate did not show significant temporal trends over a 16-year period. Bulk inputs of inorganic N were moderate, ranging from 6 to 10 kg N ha(-1) year(-1) depending on the time period considered and the degree of site exposure to polluted air masses from the Barcelona conurbation. Large dry-sedimented particles played a minor role, since wet-only inputs were virtually identical to bulk inputs. On the contrary, branch- and plate-washes indicated substantial dry inputs of N gases and small particles. Total atmospheric deposition was estimated at 15-22 kg N ha(-1) year(-1), most of it being retained within the studied broadleaved evergreen forests. Ecosystem N availability is thus likely to be increasing in these forests. PMID- 11939284 TI - Nitrate leaching in important crops of the Valencian Community region (Spain). AB - In the Valencian Community there are many areas in which the nitrate content of groundwater is higher than the limit of 50 mg l(-1) established by the European Union. A recent survey of drinking water quality showed that around 8% of the Valencian Community population had water supplies with nitrate content above that limit. The Valencian Community has an intensive agricultural base that includes citrus trees, fruit trees, and vegetables. A summary of the available nitrate leaching results in vegetable fields and citrus orchards is presented. It is demonstrated in the main vegetables grown, that N inputs were much higher than the values recommended by some researchers, and that nitrate leaching values were in most cases within the range of 150-300 kg N ha(-1). Artichoke, early potato, and onion were the three crops with higher leaching rates than other crops, based on simulation studies made for several of the main vegetable growing areas of the Valencian Community. The mineral N content in the 0-60 cm layer at planting time in the vegetable fields is, in many cases, greater than 200 kg ha(-1); fertilizer application could be greatly reduced if some recommendation system similar to the N(min) system used in other European countries was adopted. A simplified N balance in onion and early potato crops showed that the lower limit for mineralization in the 0-60 cm soil layer during the crop period varied from 65 to 130 kg N ha(-1). In these two crops nitrate leaching varied from 240 to 340 kg N ha(-1), depending on the N fertilizer treatment, and representing about 66-70% of total N input in the onion crop, and from 38 to 65% in the potato crop. Nitrate leaching losses in citrus orchards were, in general, lower than 100 kg N ha(-1) year(-1), representing about 33% of the total N input. A comparison of these results with those obtained in other studies is made. Finally, prospects for improvements in the fertilizer practice of vegetable crops and citrus orchards in the Valencian Community are discussed. PMID- 11939285 TI - Limits to nitrogen use on grassland. AB - Data from nitrogen (N) response experiments on grassland in Belgium and the Netherlands were analysed with the help of a descriptive crop N response model, to identify permissible doses below which no accumulation occurs of residual mineral soil N in autumn. N(min). Using different years as separate sets, a total of 29 data sets were obtained from eight locations on various soil types. A large variation was found in N(min) base levels (unfertilised) between locations and between years at a given location. For doses low enough not to affect crop N recovery, every 100 kg N applied was associated with 3-4 kg residual N(min) in autumn. This is considered very low compared to N(min) base levels, but values differed significantly from zero. After normalising N-doses from different sources (mineral fertiliser and cattle slurry) with the help of a coefficient expressing effectiveness based on crop N uptake, no difference was found between fertiliser and slurry in terms of their effect on residual Nmin. The above also holds for nitrate leaching as measured. The sources do differ, however, with respect to long-term effects and these are quantified with a first-order approximation. It it shown that, also after incorporation of long-term effects, much higher N-doses on grassland are justified than the 170 kg N per ha per year in animal manures currently proposed by the European Commission. On normal productive cut grassland as in the analysed experiments, total N doses in cattle slurry up to 400 kg per ha per year have very little effect on residual N(min), if not accompanied by high fertiliser doses. Introducing limits to the use of animal manures on grassland without limiting the input of mineral fertiliser-N lacks any scientific ground. PMID- 11939286 TI - Nitrogen sources to watersheds and estuaries: role of land cover mosaics and losses within watersheds. AB - Across most of the World's coastal zone there has been a geographic transition from naturally vegetated to human-altered land covers, both agricultural and urban. This transition has increased the nitrogen loads to coastal watersheds, and from watersheds to receiving estuaries. We modeled the nitrogen entering the watershed of Waquoit Bay, Massachusetts, and found that as the transition took place, nitrogen loads to watersheds increased from 1938 to 1990. The relative magnitude of the contribution by wastewater, fertilizers, and atmospheric deposition depends on the land cover mosaics of a watershed. Atmospheric deposition was the major input to the watershed surface during this period, but because of different rates of loss within the watershed. wastewater became the major source of nitrogen flowing from the watershed to the receiving estuaries. Atmospheric deposition prevails in watersheds dominated by natural vegetation such as forests, but wastewater may become a dominant source in watersheds where urbanization increases. Increased nitrogen loads resulting from conversion of natural to human-altered watershed surfaces create eutrophication of receiving waters, with attendant changes in water quality, and marked shifts in the flora and food webs of the affected estuaries. Management efforts for restoration of eutrophied estuaries require maintenance of forested land, and control of wastewater and fertilizer inputs, the major terms in most affected places subject to local management. Wastewater and fertilizer nitrogen derive from within the watershed, which means local measures may effectively be used to control eutrophication of receiving waters. PMID- 11939287 TI - Linkage of a geographical information system with the gleams model to assess nitrate leaching in agricultural areas. AB - The GLEAMS (Groundwater Loading Effects of Agricultural Management Systems, Version 2.10) model was linked with a Geographical Information System (GIS) to evaluate the risk of nitrate pollution in an area of 230 km2 near Valencia (Spain). Under Mediterranean conditions, GLEAMS was calibrated and validated using results from field experiments on citrus orchards and vegetables grown in that area. A graphical user interface (GUI) was implemented in the GIS-model system to allow a non-specialist to run the model, and to represent simulation results as a thematic map. In order to execute the GLEAMS model, data must be grouped in five basic layers: four layers correspond to the base maps (soils, climate, land use, NO3 content in irrigation water), and the fifth layer corresponds to agricultural management practices, introduced in the system interacting to the GUI. To illustrate system capabilities. two rotations with crops in the vegetable area (potato/lettuce/onion/cauliflower, and artichoke/artichoke in 2 years rotation each), and orange trees in the citrus area, were simulated to determine the N leached in the study area. Pollution risk maps show that vegetable crops and areas irrigated with groundwater have the highest potential risk of groundwater nitrate pollution. Further analysis identified potato and artichoke (in the first year) to be the crops with the higher risk. PMID- 11939288 TI - Atmospheric nitrogen deposition on the east coast of Spain: relevance of dry deposition in semi-arid Mediterranean regions. AB - Bulk deposition composition and pine branch washing were measured from April 1999 to March 2000 on the east coast of Spain. The main objective was to characterise N deposition patterns with special emphasis on dry deposition. Bulk deposition in the region is dominated by neutralisation processes by Ca2+ and HCO3-, ClNa of marine origin and a high correlation between NO3- and SO4(2-). SO4(2-) concentrations show a decrease with respect to previous studies in the region in agreement with generalized sulfur emission decreases while the remaining ions, including NO3-, are higher due to their general increase as well as to the inclusion of dry deposition in bulk collectors in the present study. An enrichment in NO3- has been observed in dry deposition composition branch washing) with respect to bulk deposition, while an impoverishment has been observed in the case of NH4+. Annual bulk deposition varies between 7.22-3.1 and 3.5-1.8 Kg ha(-1) year(-1) for S- SO4(2-) and N- NO3-, respectively. N total deposition goes from 9.78 to 6.8 Kg ha(-1) year(-1) at most stations, with the lowest deposition at the control station and Alcoi. The relative dry deposition with respect to the total was over 40% at most stations, going up to 75% at the southern station. N-deposition is expected to be higher considering that N-NH4+ deposition has been underestimated in this study. PMID- 11939289 TI - An integrative analysis of the role of atmospheric deposition and land management practices on nitrogen in the US agricultural sector. AB - Additions of anthropogenic nitrogen (N) compounds constitute one of the major classes of air pollutants of significance to human health and the environment. Reliance on wet deposition measurements alone can lead to considerable underestimates (by 40-60%) of the total (wet + dry) atmospheric N deposition. In addition, wet deposition of N are about 20% of the levels that are lost due to volatilization (primarily ammonia). Nevertheless, in the agricultural sectors of the Mississippi River basins, farm management practices, and recycling of N within cropping systems clearly outweigh the contributions of atmospheric deposition. As opposed to native vegetation and forests, there are no records of the negative effects of atmospheric N deposition on crop yield. Similarly, field studies on the interactions of atmospheric N compounds with the incidence and spread of pathogens does not permit any generalizations. Nitrogen applied as fertilizer affects disease probably more by its effect on the plant growth than by its effects on pathogens. In contrast, atmospheric nitrogen dioxide appears to be a stimulant of aphid performance. Under conditions of heavy weed infestation, N fertilization stimulates weed growth and competitiveness, rather than crop yield. PMID- 11939290 TI - Metal accumulation in mussels of the Kuril Islands, north-west Pacific Ocean. AB - Concentrations of cadmium, copper, iron, lead, manganese, nickel and zinc were investigated in two species of mussel from the Kuril Islands in the north-western Pacific Ocean: the short-lived mussel Mytilus trossulus and the much larger and longer-lived Crenomytilus grayanus. The concentrations of most elements were low, and these reported levels are believed to reflect background values for pristine locations. However, both iron and zinc concentrations were elevated in some samples taken from areas of hydrothermal vent activity, and these are believed to reflect the enrichment of the two metals in solution at such sites. Furthermore, the concentrations of cadmium were unusually elevated in both species of mussel, especially in the longer-lived C. grayanus. The high accumulation of cadmium in mussels from the Kuril Islands is believed to reflect regional upwelling, but it is also postulated that filtration rates of the mussels are high due to low levels of suspended matter, and this leads to a high assimilation efficiency for cadmium. The distribution of cadmium amongst the tissues of C. grayanus is also reported, and the need for further ecotoxicological studies in the area is proposed. PMID- 11939291 TI - Cadmium detoxification processes in the digestive gland of cephalopods in relation to accumulated cadmium concentrations. AB - The high concentrations of cadmium recorded in the digestive gland of cephalopods from various temperate and subpolar waters suggest that these molluscs have developed efficient cadmium detoxification mechanisms. The subcellular distribution of cadmium in the digestive gland cells was investigated in seven cephalopod species from the Bay of Biscay (France) and the Faroe Islands. In most species, cadmium was mainly found in the cytosolic fraction of the digestive gland cells, reaching up to 86% of the total cadmium for the squid Loligo vulgaris from the Bay of Biscay. But species with the highest total level of cadmium showed a higher percentage of cadmium associated to insoluble compounds. The quantification of metallothioneins (MTs) by the polarographic method was performed in order to evaluate the involvement of these proteins in the detoxification of the high amounts of bioaccumulated cadmium. Metallothionein levels in cephalopods ranged form 742 +/- 270 to 3478 +/- 1572 microg/g wet weight. No relationship could be established between total cadmium, cytosolic cadmium and MT levels suggesting the occurrence of other Cd-binding ligands. Although these proteins have not been characterised, as cadmium in the digestive gland of cephalopods is mainly associated with soluble ligands, a high potential transfer to predators can be predicted. PMID- 11939292 TI - DNA damage and apoptosis in the mussel Mytilus galloprovincialis. AB - The effects of known genotoxic substances (4-nitroquinoline-N-oxide, benzo[a]pyrene, teniposide, etoposide, cycloheximide, tributyltin) on human cells (FLC, HL-60) and on mussels were investigated. The correlations between formation of DNA strand breaks and DNA fragmentation characteristic for the process of apoptosis were estimated. Strand breaks induced by 4-nitroquinoline-N-oxide and benzo[a]pyrene did not correlate with DNA fragmentation detected in the process of apoptosis. Induction of internucleosomal DNA fragmentation in HL-60 cells was initiated by teniposide, etoposide and tributyltin, while in the gills of mussels this was detected only with tributyltin. Levels of DNA strand breaks in natural mussel populations, living at locations under the influence of urban and industrial wastes, do not mirror the apoptotic processes. PMID- 11939293 TI - Metallothionein in liver of eels Anguilla anguilla from the Thames Estuary: an indicator of environmental quality? AB - The purpose of this study was to examine the role of the cysteine-rich protein metallothionein (MT) in the detoxification and metabolism of metals in livers of eels Anguilla anguilla from the Thames Estuary, and to assess the value of MT measurements in environmental monitoring. Hepatic MT levels and associated metal concentrations were determined in A. anguilla collected on three occasions during 1998 at sites along the tidal Thames (from freshwater to the sea) and also from a reference site (Tamar Estuary) in southwest England. MT was present at basal levels of 2 mg g(-1) dry weight (dw), predominantly in association with the essential metals Cu and Zn. MT concentrations were variable within each eel 'population' but were generally highest (up to 11 mg g(-1) in individual eels) at the more contaminated upper- and mid-estuarine sites in the Thames (e.g. Brentford and Kew). Lowest MT levels were those in eels from Blythe Sands at the mouth of the estuary. Season, sex, reproductive status and salinity may have influenced MT levels to some extent, although metals were the most significant factor. Hepatic MT concentrations were highly correlated with associated metal burdens (Zn, Cu, Ag and Cd),--notably reflecting Cu and Ag enrichment in the upper- and mid-estuary. With the exception of Zn, metals in the supernatant fraction of eel livers were primarily associated with MT. Proportions of Cu, Ag and Cd bound to MT increased as a function of accumulated metal burdens, with no indication of saturation. Thus, despite causing induction of MT, excess bioavailable Cu, Ag and Cd appear to be successfully detoxified in eels over the range of environmental contamination encountered along the Thames Estuary. Paradoxically, it may be argued from conventional doctrine that the presence of raised levels of MT, at upstream sites, constitutes a response to contamination and, by definition, signals that the fish are affected by metals; however, obvious signs of deleterious effects were not detected. Superficially, from liver MT data, it would seem that eel populations have become biochemically adapted to metal contamination. The approach tested here has proved viable in helping to interpret trends in bioavailability and the toxicological significance of accumulated metal burdens in Thames eels. A. anguilla is a common inhabitant of estuarine and fresh waters throughout Europe, and related species occur world wide. Therefore, determination of hepatic MT (and associated metals) in eels could be useful for monitoring responses to metal exposure and environmental quality on a much broader basis. Ideally these determinations should be performed in tandem with other sublethal-effects measurements to maximise information. PMID- 11939294 TI - Alteration of the biochemical properties of female gonads and vitellins in the clam Mya arenaria at contaminated sites in the Saguenay Fjord. AB - Vitellins (Vn) are the major egg yolk proteins that constitute an important energy reserve for mollusc embryos. The purpose of the present study was to examine whether the relative levels of sugars, lipids, phosphates, and labile IIb metals and calcium normally associated with Vn would differ in clam populations living at contaminated sites. Softshell clams (Mya arenaria) were collected at three sites in the area of the Saguenay Fjord: a marina, a municipal sewer outfall zone, and a reference site. The condition factor (weight:length ratio), metallothionein-like proteins, cytochrome P450 1A1 activity and DNA damage were all determined in the clam's digestive gland. Levels of total sugars, lipids, alkali-labile phosphates, proteins, and labile zinc and calcium were determined in female gonad homogenates and in purified Vn. The results show that clam gonads at the contaminated site by a marina displayed a lower index of gonad activity than the reference site. In addition, the condition factor was significantly lower at the marina site as compared to the reference site, with a concomitant increase in DNA damage and metallothionein (MT) induction. In fact, the condition factor was significantly correlated with DNA damage (R = -0.413, P = 0.045) and MT levels (R = -0.622, P = 0.03). Homogenates of female gonads were found to contain higher levels of labile IIb metals and calcium, with lower lipid content at the marina site compared to the reference site, and much higher levels of alkali-labile phosphates (ALP) and calcium at the municipal outfall site. Vn from the marina site were significantly higher in labile IIb metals but lipid content appeared to be somewhat lower than at the reference site. Vn from the municipal site were found to be highly phosphorylated, with higher levels of lipids and calcium. These results suggest that the chemical composition of the gonads and Vn are altered in contaminated sites. PMID- 11939295 TI - Hydrocarbon effects on fouling assemblages: the importance of taxonomic differences, seasonal, and tidal variation. AB - Effects of hydrocarbon-contaminated substrata on recruitment of three species of fouling organisms were studied along the Louisiana gulf coast. Clay tiles (232 cm2) were exposed to crude oil, 10% water soluble fraction of crude oil, or 25 g/kg artificial seawater, and placed out at two locations, in two seasons, and at two tidal levels in an estuary near Port Fourchon, Louisiana. Bryozoan (Membranipora savartii) recruitment was significantly reduced in all experiments on crude oil-exposed tiles. However, oysters (Crassostrea virginica) and barnacles (Balanus eburneus) exhibited recruitment facilitation, and oysters grew to larger size, on crude oil-exposed tiles in 1-4 of the five experiments. When oyster larvae were exposed to the same treatments in the laboratory, settlement was, however, significantly depressed on crude oil-exposed tiles as compared with controls, although oyster size was larger on crude-oil exposed tiles. Recruitment on tiles exposed to the water soluble fraction of crude oil was similar to control tiles in nearly all experiments for all taxa. We suggest naturally occurring biofilms (which hydrocarbons facilitate) may promote or inhibit recruitment, depending on the taxon, because hydrocarbons facilitated recruitment only in field experiments, not in lab experiments without biofilms. However, stronger currents in the field experiments may have more rapidly diluted hydrocarbons, and hydrocarbon effects were not large in comparison with natural seasonal and tidal variation in recruitment. PMID- 11939296 TI - Prevalence of filarial antigenaemia in Papua Nnew Guinea: results of surveys by the School of Public Health and Tropical Medicine, James Cook University, Townsville, Australia. AB - During the period from 1991 to 1997 the School of Public Health and Tropical Medicine, James Cook University carried out filariasis surveys in several parts of Papua New Guinea using the newly introduced Onchocerca gibsoni monoclonal (Og4C3) and immunochromatographic test (ICT) antibody-based assays for filarial antigen and, in some cases, a Knott's test for microfilariae. The average prevalence of filarial antigenaemia and microfilaraemia was 56% and 35% respectively confirming earlier survey results that filariasis is hyperendemic in many parts of the country. The antigen tests detected 25% more cases than the Knott's test and the simplicity of the ICT and its capacity to produce almost instant results make it an ideal tool for surveys. PMID- 11939297 TI - Wuchereria bancrofti infection and disease in a rural area of Papua New Guinea. AB - The relation between infection and disease in lymphatic filariasis is still imperfectly understood. This paper presents baseline data on microfilaraemia, oedema and acute episodes from a drug trial against bancroftian filariasis in Papua New Guinea. Among 2,187 people with data on these manifestations, 1,144 were microfilaraemic, 78 had leg oedema, 356 had acute episodes and 919 were negative for all three. All possible combinations of the three manifestations were observed. The relationships between microfilaraemia and disease are found to be time-dependent. Microfilaraemia is a positive risk factor for leg oedema at lower ages, but a negative one at higher ages. Acute disease over a whole year is found to be positively associated with a point estimate of microfilaraemia. However, when considering incidence within a month of the time of the blood sample, a curvilinear relationship is found and people with low but positive microfilarial counts have the lowest incidence. Possible explanations for these findings are discussed. PMID- 11939298 TI - Towards eliminating lymphatic filariasis in Papua New Guinea: impact of annual single-dose mass treatment on transmission of Wuchereria bancrofti in East Sepik Province. AB - The impact of annual single-dose community-wide treatment on the transmission of Wuchereria bancrofti was investigated in 5 villages in the East Sepik Province where pretreatment prevalence of microfilaraemia ranged from 34% to 73%. Anopheles punctulatus and An. koliensis were the only carriers of the parasite. 3 villages received diethylcarbamazine citrate (DEC) in combination with ivermectin (IVR) and 2 received DEC alone. The rate and intensity of microfilaraemia were both reduced in all 5 villages. Reduction in prevalence was between 43% and 67% in the DEC+IVR study villages and between 24% and 27% in the DEC alone villages. Density was reduced by between 81% and 95% in the DEC+IVR villages and between 69% and 74% in the DEC alone villages. Breaks in perennial transmission (failure to detect infective mosquitoes in four or more consecutive monthly collections) occurred in all 3 communities treated with DEC+IVR. Transmission was almost completely interrupted in 2 villages, where infective mosquitoes were not detected during 11 of the 12 months following treatment. We concluded that repeated annual single-dose community-wide treatment with DEC+IVR could lead to complete interruption of transmission and ultimately elimination of lymphatic filariasis. PMID- 11939299 TI - Community empowerment in the control of lymphatic filariasis in Misima, Milne Bay Province using diethylcarbamazine in combination with albendazole. AB - We report the successful implementation of a community-based lymphatic filariasis control program using annual single-dose treatment with diethylcarbamazine (DEC) in combination with albendazole. The target population included over 28,000 people in the Samarai Murua District, Milne Bay Province, Papua New Guinea. A community-based delivery model was as effective as the standard health services delivery model. The number of people tested in 1998 before mass drug administration (MDA) and in 1999, one year after treatment, were 1644 and 942 respectively; the number who received mass treatment was 29,883 in 1998 and 28,965 in 1999. The prevalence of antigenaemia decreased significantly from 19% to 12%. The cost of running the program also decreased by 50%. The total number of trained health staff required to conduct the MDA program declined from 62 in 1998 to 12 in 1999, a reduction of 81%, with a cost saving in salary and allowances. A salient organizational initiative that surfaced was the use of local expertise in the private sector as a catalyst for obtaining funds from external sources to manage and facilitate the program which was conducted with locally available resources. PMID- 11939300 TI - The epidemiology and control of lymphatic filariasis on Lihir Island, New Ireland Province. AB - Clinical, parasitological and entomological surveys performed in 9 villages on Lihir Island, Papua New Guinea, before mass treatment with diethylcarbamazine (DEC), showed that lymphatic filariasis, caused by nocturnally periodic Wuchereria bancrofti, was endemic in 8 of them. Blood samples from 593 people revealed an overall microfilarial carrier rate of 24%. Amongst endemic villages, microfilarial carrier rates ranged from 5% to 43% and there was no significant difference in parasite prevalence between males and females. Obstructive filarial disease, defined as lymphoedema of the limbs or hydrocele, was observed in only 2% of 262 males examined. None of the 265 females examined had clinical symptoms. Entomological surveys yielded a total of 4095 mosquitoes including 3,692 anophelines and 241 culicines but only Anopheles farauti was found to harbour infective larvae of W. bancrofti. Pretreatment infection and infective rates of An. farauti were 7% and 1% respectively and up to 12 infective larvae were found in a single specimen. The microfilarial carrier rate in a cohort of people who received two DEC treatments dropped from 59% to 32% but the difference was not statistically significant. However, density of microfilaraemia decreased significantly from 170 to 10 mf/ml. Biannual mass treatment with DEC significantly reduced vector infection rates and transmission intensity on Lihir. PMID- 11939301 TI - Control of lymphatic filariasis in a hunter-gatherer group in Madang Province. AB - Diethylcarbamazine (DEC) has been successfully administered to millions of people in established villages and towns, but little or no information exists on the use of this drug to control lymphatic filariasis in isolated seminomadic groups. We have studied the impact of biannual single-dose mass treatment to control filariasis in the Hagahai, an isolated hunter-gatherer, shifting horticulturist group in the fringe highlands of Papua New Guinea. Despite low treatment coverage, 6 mass treatment rounds significantly reduced the overall prevalence of infection with Wuchereria bancrofti, by antigen detection assay, from 55% before treatment to 34% after treatment. Obstructive filarial disease in the form of elephantiasis or hydrocele was not observed among the indigenous population. Anopheles species accounted for 91% of human-biting mosquitoes collected in the area. A total of 1126 mosquitoes were caught and dissected individually but none was infected with third-stage larvae (L3). Our findings support the phenomenon of facilitation, which predicts that Anopheles-transmitted lymphatic filariasis can be interrupted by mass chemotherapy alone in areas of low vector density and low transmission intensity as observed in the Hagahai. PMID- 11939302 TI - Human immune responses to lymphatic filariasis in Papua New Guinea. AB - The following review highlights our current knowledge about the host immune response to lymphatic filariasis. Our understanding of how the host immune response influences the risk of developing disease has changed dramatically over the past decade. Previously the spectrum of disease associated with lymphatic filariasis was largely attributed to the nature of the host immune response. Now we appreciate that the duration and intensity of infection and possibly the direct influence of parasite-derived molecules also determine the risk of disease. The review highlights recent studies examining the influence of transmission intensity on the host immune response, and how this might be mediated by defects in antigen-presenting cell function, and also the role of basophils and mast cells. PMID- 11939303 TI - Efficacy of mass single-dose diethylcarbamazine and DEC-fortified salt against bancroftian filariasis in Papua New Guinea six months after treatment. AB - The efficacy of two diethylcarbamazine (DEC) treatment strategies to control bancroftian filariasis, diethylcarbamazine-fortified salt (DEC-FS) and a single DEC dose on mass administration, was evaluated in two communities in Papua New Guinea with pretreatment antigen prevalence of 55% and 71%. In the first community 0.2% w/w diethylcarbamazine-fortified salt was distributed monthly to accepting households at no cost for 12 months. In the second community a single DEC dose based on body size but designed to give about 6 mg/kg was administered to eligible acceptors. Despite wide variation in antigen prevalence among study villages there were marked reductions in prevalences under both treatment strategies. Among individuals antigenaemic on day 0, DEC-FS and a single DEC dose gave filaria antigen clearance rates of 43% and 13%, respectively. In the salt treated community the incidence of antigenaemia after 6 months in acceptors from households that received 5 kg or more of DEC-FS was 14% whereas in those receiving less than 5 kg salt was 4%. The incidence rates in the second community in those that received < 2.5 and > or = 2.5 tablets were 16% and 8%, respectively. The two treatment strategies were simple to manage and appropriate for developing countries and were widely accepted. DEC-FS was more efficacious than single-dose DEC tablets but a single administration of DEC tablets is easier to administer. PMID- 11939304 TI - Global influenza laboratory on the horizon. PMID- 11939305 TI - Serum lactoferrin concentrations in colostrum-fed calves. AB - OBJECTIVE: To determine serum lactoferrin concentrations (SLFC) in neonatal calves before and after ingestion of colostrum and to develop models that predict SLFC as a function of colostral lactoferrin concentrations (CLFC) in calves. ANIMALS: 13 Holstein calves. PROCEDURE: Calves were fed 4 L of colostrum via oroesophageal feeder within 3 hours after birth. Serum samples were collected before ingestion of colostrum (day 0) and 2, 4, 6, and 7 days after birth. Colostrum and serum IgG concentrations were measured by use of radial immunodiffusion. The CLFC and SLFC were determined by use of an ELISA. RESULTS: Mean +/- SD SLFC on days 0, 2, 4, 6, and 7 were 2.5+/-1.6 (range 0.47 to 71), 6.0+/-3.0 (range 2.0 to 16.6), 12.0+/-12.4 (range 0.0 to 43.5), 171+/-13.6 (range 2.2 to 39.4), and 13.6+/-16.4 (range 0.0 to 43.8) mg/ml, respectively. The SLFC on days 6 and 7 differed significantly from SLFC on day 0. The model that best estimated SLFC on day 6 predicted that (SLFC)2 was a function of the logarithm of relative efficiency of passive transfer (REPT) and ([CLFC]2 x [REPT]2), where R2 = 0.4. The model for SLFC on day 7 predicted that (SLFC)2 was a function of log(REPT), where R2 = 0.44. CONCLUSIONS AND CLINICAL RELEVANCE: Definitive evidence for passive transfer of lactoferrin via colostrum is lacking, because SLFC on day 2 or 4 were not significantly different than day 0. Relative efficiency of lactoferrin absorption was directly related to SLFC on day 6 but inversely related to SLFC on day 7. PMID- 11939306 TI - Frequency of the mutant MDR1 allele associated with ivermectin sensitivity in a sample population of collies from the northwestern United States. AB - OBJECTIVE: To determine the frequency of the MDR1 gene mutation (polymorphism) associated with ivermectin sensitivity in a sample population of Collies in Washington and Idaho. ANIMALS: 40 healthy client-owned Collies. PROCEDURE: A blood sample (8 ml) was collected from each dog and used for RNA extraction. Reverse transcriptase was used to generate MDR1 cDNA. Polymerase chain reaction (PCR) primers were designed to amplify a 1,061-base pair region of the MDR1 gene. The PCR products were sequenced to determine whether the Collies had 0, 1, or 2 mutant alleles. Pedigrees of some dogs were available for analysis to determine relatedness of affected dogs. RESULTS: Of the 40 Collies, 9 (22%) were homozygous for the normal allele (normal), 17 (42%) were heterozygous (carrier), and 14 (35%) were homozygous for the mutant allele (affected). Pedigree analysis revealed that some, but not all, affected dogs were related to each other within the 4 most recent generations. CONCLUSIONS AND CLINICAL RELEVANCE: A high percentage of a sample population of Collies in Washington and Idaho are affected or carriers of the mutant MDR1 allele associated with ivermectin sensitivity. A similar frequency of this mutation may be detected in dogs from other geographic areas. Pharmacologic treatment with ivermectin, loperamide, vincristine, and other drugs that are substrates of P-glycoprotein, the MDR1 gene product, may result in neurologic toxicosis in a high percentage of Collies. PMID- 11939307 TI - Calculation of variables describing plasma nonvolatile weak acids for use in the strong ion approach to acid-base balance in cattle. AB - OBJECTIVE: To calculate values for the total concentration of nonvolatile weak acids (Atot) and the effective dissociation constant for nonvolatile weak acids (Ka) of bovine plasma and to determine the best method for quantifying the unmeasured strong anion concentration in bovine plasma. SAMPLE POPULATION: Data sets from published and experimental studies. PROCEDURE: The simplified strong ion model was applied to published and experimentally determined values for pH, PCO2, and strong ion difference (SID+). Nonlinear regression was used to solve simultaneously for Atot and Ka. Four methods for quantifying the unmeasured strong anion concentration in plasma (anion gap, the Fencl base excess method [BEua], the Figge unmeasured anion method [XA], and the strong ion gap [SIG]) were compared in 35 cattle with abomasal volvulus. RESULTS: For bovine plasma at 37 C, Atot was 25 m M/L, equivalent to 76 times the albumin concentration or 3.6 times the total protein concentration; Ka was 0.87 x 10(-7), equivalent to pKa of 706. The Atot and Ka values were validated, using data sets from in vivo and in vitro studies. Plasma unmeasured strong anion concentration was most accurately predicted in critically ill cattle by calculating SIG from serum albumin (R2, 0.66) or total protein concentration (R2, 0.60), compared with BEua (R2, 0.56), [XA] (R2, 0.50), and the anion gap (R2, 0.41). CONCLUSIONS AND CLINICAL RELEVANCE: Calculated values for Atot, Ka, and the SIG equation should facilitate application of the strong ion approach to acid-base disturbances in cattle. PMID- 11939308 TI - Influence of long-term treatment with tetracycline and niacinamide on antibody production in dogs with discoid lupus erythematosus. AB - OBJECTIVE: To evaluate the effect of long-term treatment with tetracycline and niacinamide on antibody production in dogs by measuring postvaccinal serum concentrations of antibodies against canine parvovirus and canine distemper virus. ANIMALS: 10 dogs receiving long-term treatment with tetracycline and niacinamide (treatment group) and 10 healthy dogs (control group). PROCEDURE: The treatment group included 9 dogs with discoid lupus erythematosus and 1 dog with pemphigus foliaceus on long-term treatment (> 12 months) with tetracycline and niacinamide. The control group included 10 healthy dogs with no clinical signs of disease and no administered medications for the past 3 months. Blood samples were obtained from all dogs by jugular venipuncture. Serum antibody titers against canine parvovirus and canine distemper virus antigens were measured, using hemaglutination inhibition and serum neutralization, respectively, and compared between groups. RESULTS: A significant difference in antibody titers between treatment- and control-group dogs was not found. All dogs had protective antibody titers against canine distemper virus, and 8 of 10 dogs from each group had protective titers against canine parvovirus infection. CONCLUSION AND CLINICAL RELEVANCE: These results provide evidence that long-term treatment with tetracycline and niacinamide does not interfere with routine vaccinations and thus does not seem to influence antibody production in dogs. PMID- 11939309 TI - Finasteride-induced prostatic involution by apoptosis in dogs with benign prostatic hypertrophy. AB - OBJECTIVE: To determine the effect of finasteride on programmed cell death (apoptosis) of prostatic cells during prostatic involution in dogs with benign prostatic hypertrophy (BPH). ANIMALS: 9 dogs with BPH. PROCEDURE: Dogs were randomly assigned to treatment or control groups. Treatment dogs (n = 5) were administered finasteride (0.1 to 0.5 mg/kg, PO, q 24 h) for 16 weeks, whereas the 4 control dogs were administered an inert compound. Prostatic cells from the prostatic fluid portion of the ejaculate of treatment and control dogs were obtained before and 1, 2, 3, 4, 8, and 16 weeks after initiation of treatment. Cells were concentrated by use of centrifugation. Prostatic cells were examined for indications of apoptosis by use of a terminal deoxyribonucleotidyl transferase-mediated deoxyuracil triphosphate nick-end labeling technique. After receiving the inert compound for 16 weeks, the 4 control dogs were administered finasteride for 16 weeks, and evaluations were repeated. RESULTS: Percentage of apoptotic prostatic cells in ejaculated prostatic fluid of treatment dogs increased significantly (from 9% before treatment to 33, 31, 26, and 27% after 1, 2, 3, and 8 weeks of treatment, respectively). There was no significant change in percentage of apoptotic prostatic cells in the ejaculated prostatic fluid of control dogs. CONCLUSIONS AND CLINICAL RELEVANCE: Finasteride-induced prostatic involution appears to be via apoptosis in dogs with BPH. Finasteride treatment of dogs with BPH causes prostatic involution by apoptosis rather than necrosis. PMID- 11939310 TI - Serologic evaluation of five unvaccinated heifers to detect herds that have cattle persistently infected with bovine viral diarrhea virus. AB - OBJECTIVE: To determine whether serologic evaluation of 5 unvaccinated 6- to 12 month-old heifers is a valid method for identifying herds that contain cattle persistently infected (PI) with bovine viral diarrhea virus (BVDV). ANIMALS: 14 dairy herds with a history of BVDV infection, with health problems consistent with BVDV infection, or at risk for contracting BVDV infection. PROCEDURE: 5 unvaccinated 6- to 12-month-old heifers were randomly selected from each herd. Neutralizing antibody titers for type-I and -II BVDV were determined. A herd was classified as likely to contain PI cattle when at least 3/5 heifers had antibody titers > or = 128. Virus isolation was performed on all cattle to identify PI cattle. Genotype of isolated viruses was determined by nested multiplex polymerase chain reaction. RESULTS: 6 of 14 herds contained PI cattle. Sensitivity and specificity of serologic evaluation of 5 heifers for identifying these herds were 66 and 100%, respectively. In herds that contained PI cattle, the predominant BVDV titer in the tested heifers corresponded to the genotype of the isolated virus. CONCLUSIONS AND CLINICAL RELEVANCE: Serologic evaluation of unvaccinated 6- to 12- month-old heifers is an accurate method for identifying herds containing PI cattle. Both type-I and -II BVDV antibody titers should be determined to prevent herd misclassification. The genotype of BVDV found in PI cattle can be predicted by the predominant neutralizing antibody titers found in tested heifers. Serologic evaluation of 5 unvaccinated heifers can be used to determine whether a herd is likely to contain PI cattle. PMID- 11939311 TI - Results of clinical examinations, laboratory tests, and ultrasonography in dogs with pituitary-dependent hyperadrenocorticism treated with trilostane. AB - OBJECTIVE: To determine the efficacy of trilostane, a 3beta-hydroxysteroid dehydrogenase inhibitor, in dogs with pituitary-dependent hyperadrenocorticism (PDH). ANIMALS: 11 dogs with PDH. PROCEDURE: The initial dose of trilostane was 30 mg, PO, q 24 h for dogs that weighed < 5 kg and 60 mg, PO, q 24 h for dogs that weighed > or = 5 kg. A CBC count, serum biochemical analyses, urinalysis, ACTH stimulation test, and ultrasonographic evaluation of the adrenal glands were performed in each dog 1, 3 to 4, 6 to 7, 12 to 16, and 24 to 28 weeks after initiation of treatment. RESULTS: All dogs responded well to treatment. All had reductions in polyuria-polydipsia and panting and an increase in activity. Polyphagia decreased in 9 of 10 dogs, and 9 of 11 dogs had improvement of coat quality and skin condition. Concentration of cortisol after ACTH stimulation significantly decreased by 1 week after initiation of treatment. After treatment for 6 months, clinical signs resolved in 9 dogs. In the other 2 dogs, marked clinical improvement was reported for 1 dog, and moderate improvement was reported in the other dog. Ultrasonographically, there was a considerable change in the parenchyma and an increase in size of the adrenal glands. Adverse effects consisted of 1 dog with transient lethargy and 1 dog with anorexia. CONCLUSIONS AND CLINICAL RELEVANCE: Trilostane is an efficacious and safe medication for treatment of dogs with PDH. Additional studies in a larger group of dogs and characterization of progressive changes in adrenal glands are needed. PMID- 11939313 TI - Instrumented treadmill for measuring vertical ground reaction forces in horses. AB - OBJECTIVE: To develop and validate a novel instrumented treadmill capable of determining vertical ground reaction forces of all 4 limbs simultaneously in horses. SAMPLE POPULATION: Data obtained while a horse was walking and trotting on the treadmill. PROCEDURE: 18 piezo-electric force transducers were mounted between the treadmill frame and supporting steel platform to measure the actual forces at the corresponding bearing points. Each of the 18 sensor forces is equal to the sum of the unknown hoof forces weighted with the transfer coefficients of the corresponding force application points. The 4 force traces were calculated, solving at each time point the resulting equation system, using the Gaussian least-squares method. System validation comprised the following tests: determination of the survey accuracy of the positioning system, determination of the natural frequencies of the system, linearity test of the force transfer to the individual sensors, determination of superimposed forces with the treadmill integrated force measuring system (TiF) in a static configuration, and comparison of vertical ground reaction forces determined simultaneously by use of TiF and force shoes mounted on the forelimbs of a horse. RESULTS: Comparison between static test loads and TiF-calculated forces revealed deviations of < 1.4%. Force traces of TiF-calculated values and those recorded by use of the force shoes were highly correlated (r > or = 0.998). CONCLUSIONS AND CLINICAL RELEVANCE: This instrumented treadmill allows a reliable assessment of load distribution and interlimb coordination in a short period and, therefore, is suitable for use in experimental and clinical investigations. PMID- 11939312 TI - Measurement of the activation of equine platelets by use of fluorescent-labeled annexin V, anti-human fibrinogen antibody, and anti-human thrombospondin antibody. AB - OBJECTIVE: To investigate the potential use of fluorescent-labeled annexin V, anti-human fibrinogen antibody, and anti-human thrombospondin antibody for detection of the activation of equine platelets by use of flow cytometry. SAMPLE POPULATION: Platelets obtained from 6 Thoroughbreds. PROCEDURE: Flow cytometry was used to assess platelet activation as indicated by detection of binding of fluorescent-labeled annexin V, anti-human fibrinogen antibody, and anti thrombospondin antibody to unactivated and ADP-, collagen-, platelet activating factor (PAF)-, and A23187-activated equine platelets. Human platelets were used as control samples. Determination of 14C-serotonin uptake and release was used to assess the extent of platelet secretion. RESULTS: Anti-human thrombospondin antibody failed to bind to equine platelets. Annexin V bound to platelets activated with PAF or A23187 when platelets had undergone secretion. Anti-human fibrinogen antibody bound to ADP-, PAF-, and A23817-activated platelets, but binding was not dependent on platelet secretion. The extent of binding of anti fibrinogen antibody was less in equine platelets, compared with that for human platelets, despite maximal stimulation. CONCLUSIONS AND CLINICAL RELEVANCE: Activation of equine platelets can be detected by use of fluorescent-labeled annexin V and anti-human fibrinogen antibody but not by use of anti-human thrombospondin antibody. These flow cytometric techniques have the potential for detection of in vivo platelet activation in horses at risk of developing thrombotic disorders. PMID- 11939314 TI - Effects of racing on lymphocyte proliferation in horses. AB - OBJECTIVE: To measure the lymphocyte proliferation response in horses 12 to 16 hours after completion of a race. ANIMALS: 8 Thoroughbreds that competed in 14 races and 3 control Thoroughbreds that did not race. PROCEDURE: Horses participated in races during the late afternoon or evening. Venous blood samples were collected on a morning before a race (1 or 2 days before the race or on the day of the race), on the afternoon of a race (40 to 60 minutes after the race), and on the morning of the day after a race (12 to 16 hours after the race). Lymphocyte proliferation responses and WBC count were measured in samples obtained in the mornings. Plasma cortisol was measured in all samples. RESULTS: Lymphocyte proliferation responses were significantly reduced and WBC counts significantly increased 12 to 16 hours after a race. Plasma cortisol concentrations were significantly increased 40 to 60 minutes after a race. In samples from the control horses, lymphocyte proliferation responses, WBC counts, or plasma cortisol concentrations did not differ significantly among time periods. CONCLUSIONS AND CLINICAL RELEVANCE: A decrease in proliferative responses of circulating lymphocytes can be found as late as 12 to 16 hours after a horse participates in a race. Although the clinical consequences of these exercise-related alterations of the immune response are not yet known, managers of horses should take into account that the immune system of a horse may be affected by racing. PMID- 11939315 TI - Age-related changes in lymphocyte subsets of quarter horse foals. AB - OBJECTIVE: To characterize changes in lymphocyte subsets over time in foals from birth to 18 weeks of age, accounting for differences among individuals, and to determine the effect of overnight storage of blood samples on foal lymphocyte subset concentrations. ANIMALS: 8 healthy Quarter Horse foals from birth to 18 weeks of age. PROCEDURE: Blood samples were collected longitudinally from birth to 18 weeks of age and a CBC performed on each sample. The samples were stained for lymphocyte markers, either immediately or after overnight storage and analyzed by flow cytometry. RESULTS: Total leukocytes, total lymphocytes, and the absolute concentrations of all lymphocyte subsets increased significantly with age. The proportions of B29A+, CD21+, and-equine major histocompatability complex class-II molecule+ lymphocytes increased significantly with age. The proportion of equine (Eq) CD5+, EqCD8+, and EqWC4+ lymphocytes decreased significantly with age. Significant differences among foals were found with respect to initial concentrations with respect to initial concentrations, but not with respect to the rate of increase of the various subsets tested. Significant differences were not found in subset values when comparing blood samples stained on the day of collection or after overnight storage at room temperature (approx 21 C) or under refrigeration. CONCLUSIONS AND CLINICAL RELEVANCE: These results are consistent with an increase in subset numbers and proportions over time, but with individual differences among foals. The observation of individual differences in subsets among foals suggests that there may be individual differences in susceptibility to infectious disease during the perinatal period. The absence of an effect of overnight storage makes field studies of lymphocyte subset concentrations more feasible. PMID- 11939316 TI - Cardiovascular changes associated with intravenous administration of fumonisin B1 in horses. AB - OBJECTIVE: To determine whether cardiovascular dysfunction is evident in horses with leukoencephalomalacia experimentally induced by administration of fumonisin B1. ANIMALS: 11 healthy horses of various breeds (body weight, 252 to 367 kg). PROCEDURE: Horses were randomly assigned to 3 groups and administered fumonisin B1 daily. Horses received IV injections of 0 (control horses; n = 4), 0.01 (3), or 0.20 mg (4) of fumonisin B1/kg for 7 to 28 days. Horses were examined daily for evidence of neurologic disease. When neurologic signs consistent with leukoencephalomalacia were evident, horses were anesthetized, and catheters were inserted for evaluation of the cardiovascular system. After recovery from anesthesia, hemodynamic measurements were obtained. RESULTS: Fumonisin-treated horses with clinical signs of neurologic disease had evidence of cardiovascular dysfunction manifested as decreases in heart rate, cardiac output, right ventricular contractility (assessed by measuring the maximal rate of change of right ventricular pressure), coccygeal artery pulse pressure, and pH and base excess in venous blood as well as increases in systemic vascular resistance, compared with values for control horses. Fumonisin-treated horses with and without clinical signs of neurologic disease also had higher serum and right ventricular sphinganine and sphingosine concentrations than control horses. CONCLUSIONS AND CLINICAL RELEVANCE: An association was detected among fumonisin induced neurologic disease, increased serum and myocardial sphinganine and sphingosine concentrations, and decreased cardiovascular function in horses. Fumonisin-induced decreases in cardiovascular function may contribute to the pathophysiologic development of leukoencephalomalacia in horses. PMID- 11939317 TI - Elemental composition of teeth with and without odontoclastic resorption lesions in cats. AB - OBJECTIVE: To determine elemental composition of teeth with and without odontoclastic resorption lesions (ORL) in cats. SAMPLE POPULATION: Normal teeth from 22 cadaver cats and ORLaffected teeth from 21 cats admitted to the veterinary hospital for dental treatment. PROCEDURE: An electron microprobe was used to analyze weight percentages of calcium, phosphorus, fluorine, sodium, magnesium, sulfur, potassium, and iron in enamel, dentin, and cementum. RESULTS: Calcium and phosphorus were the most abundant elements. Fluorine, sodium, and magnesium combined were < 5% and sulfur, potassium, and iron combined were < 0.1% of total elemental composition. In enamel of normal teeth, a significant sex-by jaw location interaction was seen in mean (+/- SD) phosphorus content, which was higher in mandibular teeth of females (1764+/-0.41%) but lower in mandibular teeth of males (16.71+/-0.83%). Mean iron content in dentin of normal teeth was significantly lower in mandibular teeth than maxillary teeth (0.014+/-0.005% vs 0.023+/-0.019%). Mean enamel sodium content was significantly higher (0.77+/ 0.046% vs 0.74+/-0.025) and mean enamel iron content was significantly lower (0.017+/-0.008% vs 0.021+/-0.005%) in ORLaffected teeth, compared with normal teeth. In cementum, mean fluorine content was significantly lower (2.98%+/-0.27 vs 2.99+/-0.20%) and mean magnesium content was significantly lower (0.54+/-0.13% vs 0.60+/-0.13%) in ORL affected teeth, compared with normal teeth. CONCLUSIONS AND CLINICAL RELEVANCE: Results of our study establish baseline mineral content of enamel, dentin, and cementum for normal teeth in cats. Minimal differences in mineral content of enamel and cementum of normal and ORL-affected teeth were detected. PMID- 11939319 TI - Use of chromametry and digital photography for objective measurement of skin color in clinically normal dogs. AB - OBJECTIVE: To evaluate whether skin erythema in clinically normal dogs can be quantified by use of chromametry and image analysis of digital photographs. ANIMALS: 9 German Shepherd Dogs and 10 mixed-breed dogs. PROCEDURE: Hair was clipped at 7 sites on the body. Skin erythema was evaluated at the axillary region, right and left lateral aspect of thorax, right and left loin area (ie, part of the back between the thorax and pelvis), right and left groin area (ie, the junctional region between the abdomen and thigh), metatarsal digital pad, and on the nose. Replicate measurements were done by use of chromametry and image analysis of digital photographs, using erythema values in accordance with the Committee International d'Eclairage (CIE)-Lab color system. RESULTS: Repeatability was high for both techniques. Within-dog variation was lower than between-dog variation. Between-dog variation was high for both groups of dogs. Interregional variation was significant in German Shepherd Dogs and mixed-breed dogs. Erythema values revealed symmetry between the right and left lateral aspects of the thorax and loin and groin areas. CONCLUSIONS AND CLINICAL RELEVANCE: Precise objective methods are available for skin erythema quantification. Chromametric and photographic erythema values had a high within dog reproducibility. Between-dog variability was high for German Shepherd Dogs and mixed-breed dogs as was regional variation, indicating differences in color among dogs. PMID- 11939318 TI - Effects of equine recombinant interleukin-1alpha and interleukin-1beta on proteoglycan metabolism and prostaglandin E2 synthesis in equine articular cartilage explants. AB - OBJECTIVES: To evaluate the effects of equine recombinant interleukin-1alpha (rEqIL-1alpha) and recombinant interleukin-1beta (rEqIL-1beta) on proteoglycan metabolism and prostaglandin E2 (PGE2) synthesis by equine articular chondrocytes in explant culture. SAMPLE POPULATION: Near full-thickness articular cartilage explants (approx 50 mg) harvested from stifle joints of a 3-year-old and a 5-year old horse. PROCEDURE: Expression constructs containing cDNA sequences encoding EqIL-1alpha and EqIL-1beta were generated, prokaryotically expressed, and the recombinant protein purified. Near full-thickness articular cartilage explants (approx 50 mg) harvested from stifle joints of a 3-year-old and a 5-year-old horse were separately randomized to receive rEqIL-1alpha or rEqIL-1beta treatments 10 to 500 ng/ml). Proteoglycan release was evaluated by 1,9 dimethylmethylene blue spectrophotometric analysis of explant media glycosaminoglycan (GAG) concentration and release of 35S-sulfate-labeled GAG to explant media. Proteoglycan synthesis was assessed by quantification of 35S sulfate incorporation into proteoglycan. Explant media PGE2 concentrations were evaluated using a PGE2-specific enzyme-linked immunoassay. Data were collected at 48-hour intervals and normalized by DNA content. RESULTS: Proteoglycan release was induced by rEqIL-1alpha and rEqIL-1beta at concentrations > or =0.1 ng/ml, with 38 to 76% and 88 to 98% of total GAG released by 4 and 6 days, respectively. Inhibition of proteoglycan synthesis (42 to 64%) was observed at IL-1 concentrations > or = 0.1 ng/ml at 2 and 4 days. Increased PGE2 concentrations were observed at IL-1 concentrations > or = 0.1 ng/ml at 2 and 4 days. CONCLUSIONS AND CLINICAL RELEVANCE: The rEqIL-1 induced potent concentration dependent derangement of equine chondrocyte metabolism in vitro. These findings suggest this model may be suitable for the in vitro study of the pathogenesis and treatment of joint disease in horses. PMID- 11939320 TI - Evaluation of skin erythema by use of chromametry and image analysis of digital photographs after intradermal administration of histamine in dogs. AB - OBJECTIVE: To investigate whether the degree of erythema during an induced erythematous reaction, the histamine skin test reaction, can be assessed objectively by use of chromametry and image analysis of digital photographs. ANIMALS: 9 pet dogs (6 Golden Retrievers and 3 yellow Labrador Retrievers). PROCEDURE: Histamine phosphate was injected intradermally, and erythema of the wheal reaction was evaluated during the hour that followed. This was done by use of clinical scores, chromametry, and image analysis of digital photographs. Method reproducibility was tested for visual evaluation of printouts of digital photographs and for image analysis of the same photographs. RESULTS: The coefficient of variation of the technically derived erythema values was < 10%. The reproducibility of image analysis was high and the range of agreement between observers narrow. Using chromametry, it was not possible to differentiate between various degrees of erythema intensity as visually perceived. In contrast, use of image analysis of digital photographs enabled discrimination of slight erythema from moderate and marked erythema. The dynamics of reaction measured by chromametry followed the clinical observation. CONCLUSIONS AND CLINICAL RELEVANCE: Chromametric values are comparable to those obtained by visual inspection. As the result of standardized conditions, chromametry is preferred over digital photography. PMID- 11939321 TI - Analysis of proglycogen and macroglycogen content in muscle biopsy specimens obtained from horses. AB - OBJECTIVE: To determine proglycogen (PG) and macroglycogen (MG) content in equine skeletal muscle and to compare 2 analytical methods (acid hydrolysis [AC] and PG plus MG determination) for measurement of total muscle glycogen content (Gly(tot)) in biopsy specimens. SAMPLE POPULATION: Muscle biopsy specimens obtained from 41 clinically normal horses. PROCEDURE: Forty-five muscle biopsy specimens obtained from the middle gluteal (n = 31) or triceps (14) muscle were analyzed, using AC and MG plus PG determination for Gly(tot). Variability within muscle biopsy specimens for each method was calculated from duplicate analyses of muscle specimens. In a second experiment, variation in MG and PG content between muscle biopsy specimens and the effect of sample collection depth on the concentration of MG and PG in the middle gluteal muscle was evaluated. RESULTS: There was a strong correlation (r = 0.99) between Gly(tot) values obtained by use of AC and MG plus PG determination. Coefficients of variation for within- and between-specimen variability of Gly(tot) were approximately 4% for each method. The PG fraction was always in excess of the MG fraction. Biopsy specimens obtained from the superficial part of the middle gluteal muscle contained significantly more Gly(tot) and PG than specimens obtained from deeper parts. CONCLUSIONS AND CLINICAL RELEVANCE: This study confirms that MG and PG exist in equine skeletal muscle and can be measured reliably in biopsy samples. This technique could be applied in future studies to investigate glycogen metabolism in exercising horses and horses with glycogen-storage diseases. PMID- 11939322 TI - Analysis of synovial fluid from clinically normal alpacas and llamas. AB - OBJECTIVE: To establish reference range values for synovial fluid from clinically normal New World camelids. ANIMALS: 15 llamas and 15 alpacas. PROCEDURE: Llamas and alpacas were anesthetized with an IM injection of a xylazine hydrochloride, butorphanol tartrate, and ketamine hydrochloride combination. Synovial fluid (1 to 2 ml) was obtained by aseptic arthrocentesis from the radiocarpal and tarsocrural joints. Synovial fluid evaluation included determination of total nucleated cell count (NCC), absolute number and percentage of polymorphonuclear (PMN) and mononuclear leukocytes, total protein, and specific gravity. RESULTS: Synovial fluid evaluation revealed a total NCC of 100 to 1,400 cells/microl (mean +/- SD, 394.8+/-356.2 cells/microl; 95% confidence interval [CI], 295.2 to 494.6 cells/microl). Mononuclear leukocytes were the predominant cell type with lymphocytes, composing 50 to 90% (mean, 75.6+/-172%; 95% CI, 70.8 to 80.4%) of the mononuclear leukocytes. Approximately 0 to 12% (mean, 1.3+/-2.9%; 95% CI, 0.49 to 2.11%) of the cells were PMN leukocytes. Total protein concentrations ranged from 2.0 to 3.8 g/dl (mean, 2.54+/-0.29 g/dl; 95% CI, 2.46 to 2.62 g/dl); the specific gravity ranged between 1.010 and 1.026 (mean, 1.017+/-0.003; 95% CI, 1.016 to 1.018). CONCLUSION AND CLINICAL RELEVANCE: In llamas and alpacas, significant differences do not exist between species or between limbs (left vs right) or joints (radiocarpal vs tarsocrural) for synovial fluid values. Total NCC and absolute number and percentage of PMN and mononuclear leukocyte are similar to those of other ruminants and horses. However, synovial fluid total protein concentrations in New World camelids are high, compared with other domestic species. PMID- 11939323 TI - Comparison of effects of dexamethasone and the leukotriene D4 receptor antagonist L-708,738 on lung function and airway cytologic findings in horses with recurrent airway obstruction. AB - OBJECTIVE: To evaluate whether the leukotriene (LT) D4 receptor antagonist L 708,738 is therapeutically beneficial in treating horses with recurrent airway obstruction (heaves). ANIMALS: 12 adult horses with heaves and healthy lung lobes from 20 slaughtered horses. PROCEDURE: Lung lobes were used for smooth muscle tension and radioligand binding studies. Horses with heaves were given a placebo for 14 days and administered L-708,738 (n = 6; 2.5 mg/kg PO, q 12 h) or dexamethasone (6; 0.04 mg/kg, IV, q 24 h) from days 14 to 28. Pulmonary function was measured weekly for 36 days, and bronchoalveolar cells were collected on days 0,14, and 29 for cytologic examination. RESULTS: Nanomolar concentrations of L 708,738 were effective at antagonizing LTD4-induced bronchoconstriction and LTD4 receptor binding in lung lobes. Mean peak and trough L708,738 plasma concentrations during the treatment period were 1.54 and 0.28 microM, respectively. On days 21 and 29, lung mechanics were significantly improved in the dexamethasone-treated horses but not in the L-708,738-treated horses. Neither dexamethasone nor L-708,738 had a significant effect on cytologic findings. CONCLUSIONS AND CLINICAL RELEVANCE: L-708,738 was bioavailable after oral administration and sustained concentrations in plasma during the dosing period that exceeded in vitro efficacy values. However, airway function did not improve, suggesting that either drug concentrations in the lungs were subtherapeutic or that cysteinyl LT may not be important mediators of airway inflammation in heaves. Results provide the first evidence of cysteinyl LT1 receptors in airways of horses. PMID- 11939324 TI - Development and evaluation of an enzyme-linked immunosorbent assay for quantification of the humoral response of cattle vaccinated against Campylobacter fetus. AB - OBJECTIVE: To develop a reliable ELISA by use of a unique antigen preparation for serum IgG quantification after vaccination against Campylobacter fetus in cattle. ANIMALS: Twenty-six 24-month-old virgin Hereford heifers and a naturally infected Hereford bull. PROCEDURES: 5 antigens were prepared from a cell suspension of C fetus. Antigen preparations were the same as those reported in the literature, with the exception of antigens that were obtained by detergent solubilization of a C fetus cell suspension. For each antigen preparation, the optimal ELISA conditions for its immobilization were determined. Biotinylated antibodies against bovine immunoglobulins were obtained and used in the ELISA. Two groups of heifers were inoculated with commercial vaccines according to manufacturers' instructions. A control group was included. The immune response of vaccinated heifers and controls was followed for 6 months. RESULTS: Detergent solubilized C fetus antigens resulted in better ELISA performance than other antigen preparations. Antigens were optimally immobilized at neutral pH and low ionic strength. All antigen preparations saturated the well with the same amount of protein. The vaccination schedule that advised a booster resulted in higher antibody titers, which were sustained over a longer period than the other schedule. CONCLUSIONS AND CLINICAL RELEVANCE: In the vaccination of cattle against C fetus, the ELISA we have developed may be used to evaluate serum antibody concentrations in response to various vaccines and vaccination schedules. Our results indicate that it is advisable to include a booster in the immunization protocol. PMID- 11939325 TI - Time-dependent changes in plasma concentrations of 3-methylindole and blood concentrations of 3-methyleneindolenine-adduct in feedlot cattle. AB - OBJECTIVE: To describe time-dependent changes in plasma concentrations of 3 methylindole (3MI) and blood concentrations of 3-methyleneindolenine (3MEIN) adduct in feedlot cattle. ANIMALS: 64 yearling steers. PROCEDURES: Steers were assigned to 2 groups (32 steers/group). During the first 8 weeks, blood samples were collected from group 1 before the morning ration was fed, whereas samples from group 2 were collected 2 to 3 hours after the ration was fed. Blood samples were collected from all steers approximately 4 times/wk for 3 weeks and 3 times/wk for the subsequent 5 weeks. Samples were collected at the same time for all steers for an additional 10 weeks. Plasma samples were analyzed for 3MI concentrations. Blood samples collected from cattle in group 2 during the first 8 weeks were analyzed for 3MEIN-adduct concentrations. RESULTS: Mean blood concentration of 3MEIN-adduct increased to a maximum value on day 33 (0.80 U/microg of protein) and then decreased to a minimum on day 54 0.40 U/microg of protein). Plasma 3MI concentrations initially decreased and remained low until after day 54. Group-1 cattle had lower plasma 3MI concentrations, compared with concentrations for group-2 cattle. Blood 3MEIN-adduct concentrations and plasma 3MI concentrations were not associated with deleterious effects on weight gains. CONCLUSIONS AND CLINICAL RELEVANCE: Blood 3MEIN-adduct concentrations peaked during the period of greatest risk for development of bovine respiratory disease complex. Conversely, plasma 3MI concentrations decreased during the same period. Animal-to-animal variation in metabolic capacity to convert 3MI to 3MEIN may be of more importance than differences in plasma 3MI concentration. PMID- 11939326 TI - Concentrations of cartilage oligomeric matrix protein in dogs with naturally developing and experimentally induced arthropathy. AB - OBJECTIVE: To assay concentrations of cartilage oligomeric matrix protein (COMP) in canine sera and synovial fluid (SF), to compare COMP concentrations in clinically normal dogs and dogs with joint disease, and to analyze changes in COMP concentrations in dogs with experimentally induced acute synovitis. ANIMALS: 69 control dogs without joint disease, 23 dogs with naturally occurring aseptic arthropathy, and 6 dogs with experimentally induced synovitis. PROCEDURE: Serum (n = 69) and SF (36) were obtained from control dogs. Samples of serum (n = 23) and SF (13) were obtained from dogs with naturally occurring aseptic arthropathy with or without radiographic features of osteoarthritis (OA). Serum and SF were obtained before and 1, 2, 3, and 7 days after induction of synovitis. The COMP concentrations were determined by use of an inhibition ELISA that had canine cartilage COMP and monoclonal antibody against human COMP. RESULTS: Concentrations of COMP in serum and SF of control dogs were 31.3+/-15.3 and 298.7+/-124.7 microg/ml, respectively. In naturally occurring OA, COMP concentrations in serum (44.9+/-177 microg/ml) and SF (401.7+/-74.3 microg/ml) were significantly higher than corresponding concentrations in control dogs. The COMP concentration in SF peaked 24 and 48 hours after induction of synovitis, whereas concentration in serum peaked on day 3. CONCLUSIONS AND CLINICAL RELEVANCE: These results supported the hypothesis that COMP concentration in serum and SF of dogs may be altered after cartilage degradation or synovitis. Measurement of COMP concentrations can be useful when differentiating arthropathies in dogs. PMID- 11939327 TI - Isolation, identification, and characterization of compounds from acer rubrum capable of oxidizing equine erythrocytes. AB - OBJECTIVE: To identify compounds in Acer rubrum that cause hemolysis or oxidation of equine erythrocytes and determine whether these toxins are found in other Acer spp. SAMPLE POPULATION: Equine erythrocytes. PROCEDURE: Washed erythrocytes were incubated with extracts and fractions of Acer spp that were separated by thin layer chromatography. Methemoglobin and hemolysis were measured spectrophotometrically. Compounds within Acer spp fractions associated with cell oxidation or hemolysis were identified by gas chromatography-mass spectrometry. RESULTS: Erythrocytes incubated separately with either A. rubrum, A. saccharum, or A. saccharinum extracts had increased methemoglobin formation, compared with extract-free control samples. Two Acer spp fractions had toxic effects on erythrocytes in vitro. A major component of the Acer fraction that caused a significant amount of methemoglobin formation was identified as gallic acid. An amount of gallic acid equivalent to that found in A. rubrum extract significantly increased methemoglobin, compared with extract-free control erythrocytes, but caused less methemoglobin formation than A. rubrum extracts did. A potential co oxidant, 2,3-dihydro-3,5-dihydroxy-6-methoxy-4H-pyran-4-one, was found in the A. rubrum extract and may have been responsible for increasing methemoglobin formation. A second A. rubrum fraction caused methemoglobin formation and significant hemolysis. A. saccharum and A. saccharinum extracts caused hemolysis but less than the A. rubrum extracts did. CONCLUSION AND CLINICAL RELEVANCE: Oxidants in A. rubrum are also found in A. saccharum and A. saccharinum, and the ingestion of A. saccharum and A. saccharinum poses a potential threat to horses. PMID- 11939328 TI - Effect of vaccination on serum concentrations of total and antigen-specific immunoglobulin E in dogs. AB - OBJECTIVE: To determine the effect of vaccination on serum concentrations of total and antigen-specific IgE in dogs. ANIMALS: 20 female Beagles. PROCEDURE: Groups of 5 dogs each were vaccinated repeatedly between 8 weeks and 4 years of age with a multivalent and rabies vaccine, a multivalent vaccine only, or a rabies vaccine only. A fourth group of 5 dogs served as unvaccinated controls. Serum concentrations of total immunoglobulins and antigen-specific IgE were determined following vaccination. RESULTS: -The multivalent vaccine had little effect on serum total IgE concentrations. The concentration of IgE increased slightly following vaccination for rabies at 16 weeks and 1 year of age and increased greatly after vaccination at 2 and 3 years of age in most dogs, with a distinct variation between individual dogs. Vaccination had no effect on serum concentrations of IgA, IgG, and IgM as measured at 2 and 3 years of age. The rabies vaccine contained aluminum adjuvant in contrast to the multivalent vaccine. An increase of IgE that was reactive with vaccine antigens, including bovine serum albumin and bovine fibronectin, was detected in some of the dogs vaccinated for rabies. There was no significant correlation between serum concentrations of total IgE and antigen-specific IgE following vaccination. Serum total IgE concentration rapidly returned to preimmunization concentrations in most dogs, but high concentrations of antigen-specific IgE persisted. CONCLUSIONS AND CLINICAL RELEVANCE: Vaccination of dogs for rabies increases serum concentrations of total IgE and induces IgE specific for vaccine antigens, including tissue culture residues. Vaccination history should be considered in the interpretation of serum total IgE concentrations. PMID- 11939329 TI - Relationships between fecal consistency and colonic microstructure and absorptive function in dogs with and without nonspecific dietary sensitivity. AB - OBJECTIVE: To determine relationships between fecal consistency and colonic microstructure and absorptive function in dogs with and without nonspecific dietary sensitivity. ANIMALS: 12 dogs with nonspecific dietary sensitivity (affected) and 9 healthy dogs (controls). PROCEDURE: Affected dogs were fed 4 test diets and control dogs, 3 diets for 4 weeks each in a crossover design. Fecal consistency was assessed daily. At the end of each feeding period, electrolyte and water transport were assessed, and colonic biopsy specimens were obtained for histologic examination and measurement of crypt water uptake by use of confocal microscopy. RESULTS: Feces were consistently looser in affected dogs. In control dogs, we detected net colonic absorption of sodium and chloride and secretion of potassium and bicarbonate. Absorption of sodium and chloride was less in affected dogs, compared with controls, indicating that electrolyte transport was disrupted in affected dogs. This disruption was accentuated during feeding of diets associated with significantly poorer fecal consistency (ie, loose feces). Fecal consistency was inversely correlated with crypt water absorption, which was reduced in affected dogs. Colonic crypts were shorter and less dense in affected dogs fed diets associated with poor fecal consistency, compared with affected dogs fed other diets or with control dogs. CONCLUSIONS AND CLINICAL RELEVANCE: Colonic transport function is a major determinant of fecal consistency in dogs. Dogs with nonspecific dietary sensitivity are particularly susceptible to diet-induced changes in absorptive function. Such changes are associated with damage to colonic microstructure, disrupted electrolyte transport, and failure to dehydrate luminal contents. PMID- 11939330 TI - Evaluation of a porcine circovirus type 2-specific antigen-capture enzyme-linked immunosorbent assay for the diagnosis of postweaning multisystemic wasting syndrome in pigs: comparison with virus isolation, immunohistochemistry, and the polymerase chain reaction. AB - Quantitative virus isolation, immunohistochemistry, polymerase chain reaction (PCR) assay, and a porcine circovirus 2 (PCV2)-specific antigen-capture enzyme linked immunosorbent assay (ELISA) were used for differentiation between clinical and subclinical PCV2 infections of swine. Tissue samples from pigs experimentally infected with PCV2 and field cases of postweaning multisystemic wasting syndrome and PCV2-associated reproductive disorders were used in this evaluation. In initial studies on 6 PCV2 pools using 3 previously published PCR protocols for PCV2 detection, quantitative virus isolation, and antigen-capture ELISA, substantial differences in sensitivity were identified among these procedures. Examination of tissue samples from diseased and clinically normal pigs indicated that immunohistochemistry, quantitative virus isolation, and antigen-capture ELISA could be used to differentiate between clinical and subclinical PCV2 infections, but the PCR assay could not. Because subclinical infections of pigs with PCV2 are common, the use of nonquantitative PCR as a diagnostic tool for PCV2-related diseases should be discouraged and the PCV2-specific antigen-capture ELISA evaluated further. PMID- 11939331 TI - Confirmation of vertical transmission of bovine immunodeficiency virus in naturally infected dairy cattle using the polymerase chain reaction. AB - The purpose of this study was to determine whether bovine immunodeficiency virus (BIV) is vertically transmitted in naturally infected dairy cattle. Twenty-two dam/calf pairs from a Mississippi Agriculture and Forestry Experiment Station dairy were the study group. Blood samples were collected following delivery of calves, the peripheral blood leukocytes were purified from these samples, and the leukocyte DNA was used in polymerase chain reactions targeting the pol gene region of the BIV provirus. Southern blotting and hybridization were used to confirm the BIV specificity of the amplified fragments. BIV provirus was detected in 14 of 22 calves (64%), demonstrating vertical transmission. Eight of the calves were disqualified from the final interpretation of transplacental transfer because they may have nursed their mothers prior to blood collection, allowing the possibility of lactogenic transfer of the virus. Transplacental transmission of BIV was identified in 6 of 22 calves (27%). PMID- 11939332 TI - Detection of bovine viral diarrhea virus by TaqMan reverse transcription polymerase chain reaction. AB - Detection and elimination of calves and cows persistently infected with bovine viral diarrhea virus (BVDV) is important for the control of this pathogen. Historically, BVDV detection involved cell culture isolation followed by virus detection through immunofluorescence or immunoperoxidase monolayer assay (IPMA) methods. More recently, immunohistochemistry (IHC) has been added as a routine test for BVDV detection. The detection of BVDV by gel-based reverse transcription polymerase chain reaction (RT-PCR) is more sensitive and rapid than by cell culture isolation, but test results can be compromised by sample contamination during nucleic acid amplification. This study was designed to develop a closed tube format of BVDV nucleic acid amplification and detection, TaqMan RT-PCR. The results of this new technique were compared with those obtained with virus isolation, IPMA, and IHC. With TaqMan RT-PCR, BVDV was detected in many samples negative by IPMA, IHC, and virus isolation with the exception of 1 sample that was positive by IHC. TaqMan RT-PCR in a closed-tube format offers a rapid, economical, high volume, and sensitive method for BVDV detection without the concerns of amplified cDNA product contamination associated with open-tube gel based PCR tests. PMID- 11939334 TI - Effects of different variables on whole blood cholinesterase analysis in dogs. AB - The influence of several variables such as sample and reagent storage, anticoagulants, reaction temperature, pH, and substrate concentration on whole blood cholinesterase determination was studied. Storage of nondiluted whole blood samples at room temperature or under refrigeration (4 C) was adequate for short term storage (3 days to 2 weeks). However, freezing would be more appropriate for long-term storage (> or = 1 month), and successive thawing and freezing did not produce any loss of cholinesterase activity. All reagents (2,2'-dithiodipyridine as chromophore and acetylthiocholine and butyrylthiocholine as substrates) were stable for 3 months when frozen. Heparin and ethylenediaminetetraacetic acid were the most suitable anticoagulants for whole blood acetylcholinesterase and butyrylcholinesterase determination, because citrate yielded lower acetylcholinesterase values and fluoride inhibited butyrylcholinesterase. Increases in reaction temperature and pH yielded higher cholinesterase values but also increased nonenzymatic substrate hydrolysis. Higher cholinesterase and nonenzymatic substrate hydrolysis values were obtained as higher substrate concentrations were used. PMID- 11939333 TI - Development and application of quantitative polymerase chain reaction assay based on the ABI 7700 system (TaqMan) for detection and quantification of Mycobacterium avium subsp. paratuberculosis. AB - Numerous reports have described diagnostic methods based on the polymerase chain reaction (PCR) used to detect Mycobacterium avium subsp. paratuberculosis, the causative agent of Johne's disease. The result of conventional PCR tests has been only qualitative, either positive or negative; it does not present any quantitative information about the number of the agents in the specimen. A quantitative PCR method (IS900 TaqMan) was developed to measure the number of M. a. paratuberculosis organisms present in field and clinical samples. The sensitivity of IS900 TaqMan was 1 colony-forming unit (CFU) for M. a. paratuberculosis ATCC 19698. The specificity of the method was determined by testing 14 mycobacterial species (M. abscessus, M. asiaticum, M. avium subsp. avium, M. bovis, M. fortuitum subsp. fortuitum, M. intracellulare, M. kansasii, M. marinum, M. phlei, M. scrofulaceum, M. simiae, M. smegmatis, M. terrae, and M. ulcerans) and 9 nonmycobacterial species (Borrelia burgdorferi, Chlamydia psittaci, Ehrlichia canis, E. equi, E. risticii, Escherichia coli, E. coli O157:H7, Streptococcus equi, and S. zooepidemicus). Even at high cell numbers (10(5) CFU/reaction), most of the organisms tested negative for the IS900 insertion element except M. marinum and M. scrofulaceum. This finding for M. scrofulaceum was consistent with previous reports that several M. scrofulaceum like isolates were positive for IS900. Those isolates had 71-79% homology with M. a. paratuberculosis in the region of IS900. When used in conjunction with the new liquid medium-based ESP culture system II for bovine clinical fecal samples, IS900 TaqMan confirmed that the ESP II-positive samples contained 10(5)-10(6) CFU/ml of M. a. paratuberculosis. All of the 222 ESP II-positive and acid-fast bacilli-positive samples tested in this study were positive by IS900 TaqMan. IS900 TaqMan was also useful in the study of growth characteristics of 3 groups of M. a. paratuberculosis strains in bovine fecal samples from 3 shedding levels (heavy, medium, and low) based on cell numbers measured by Herrold egg yolk (HEY) agar culture. When cultured in ESP medium, M. a. paratuberculosis reached 10(5) 10(6) CFU/ml within 2 weeks for heavy shedders, 3-4 weeks for medium, and 6-8 weeks for low shedders. No significant growth was observed after up to 5 weeks of incubation for some of low shedders. No or extremely slow growth characteristic of low shedders might be a possible explanation for frequent false-negative results by HEY. The detection time was dependent on the inoculum size and the growth rate of M. a. paratuberculosis. Generation times were inversely proportional to the shedding level: 1-2 days for medium and heavy shedders and >4 days for low shedders. IS900 TaqMan could be a useful tool for determining viable cell counts by measuring changes in cell numbers over the incubation period. PMID- 11939335 TI - A familial degenerative neuromuscular disease of Gelbvieh cattle. AB - A degenerative skeletal muscle disease with vascular, neurologic, and renal lesions and a probable familial distribution was identified in 4-20-month-old purebred Gelbvieh cattle. Thirteen affected animals were confirmed from 6 separate beef herds, with a mortality rate of 100%. Clinical signs in affected animals consisted of ataxia, weakness, and terminal recumbency. Gross and histologic muscle lesions were indicative of nutritional myopathy of ruminants, with a lack of myocardial lesions in most cases and only rare myocardial changes in a few animals. Acute to chronic lesions in most large skeletal muscle groups consisted of degeneration, necrosis, regeneration, fibrosis, and atrophy. Fibrinoid necrosis of arterioles was a common feature in multiple tissues. Lesions in the spinal cord white matter and peripheral nerves consisted of degeneration of the dorsal columns and axons, respectively. Changes in the kidneys consisted of chronic interstitial nephritis with fibrosis, hyaline droplet change and tubular epithelial vacuolar change and were most severe in the older calves. Intracytoplasmic myoglobin and iron were demonstrated within the hyaline droplets in degenerate renal cortical tubular epithelial cells. Vitamin E levels were deficient in most (6/7) of the animals tested. Investigation of the pedigree of affected animals revealed a common ancestry for all but 1 of the animals whose parentage could be traced. This investigation suggests that a hereditary metabolic defect, possibly involving antioxidant metabolism, could be responsible for this condition. Renal disease, possibly secondary to myoglobinuria, may be unique to this bovine condition. PMID- 11939336 TI - Primary epitheliotropic intestinal T-cell lymphoma in a horse. AB - A 17-year-old Quarterhorse gelding with a clinical diagnosis of protein-losing enteropathy was submitted for necropsy following a 4-5-month duration of weight loss, decreased appetite, and hypoproteinemia. Gross findings included multiple 1 2-cm diameter ulcers on the luminal surfaces of the duodenum and ileum. Histologic examination revealed individual large, round cells infiltrating much of the mucosal epithelium of the duodenum, jejunum, ileum, and colon in addition to multifocal areas of ulceration. Similar round cells infiltrated Brunner's glands and expanded the submucosa beneath the foci of ulceration. Immunohistochemical staining indicated the round cell population was of T lymphocyte origin. Several features of this equine neoplasm bear similarities to enteropathy-associated T-cell lymphoma in humans. PMID- 11939337 TI - Antimicrobial susceptibility of Actinobacillus pleuropneumoniae, Escherichia coli, and Salmonella choleraesuis recovered from Taiwanese swine. AB - Minimum inhibition concentrations (MICs) were determined for ampicillin, ceftiofur, cephalothin, chloramphenicol, enrofloxacin, gentamicin, lincomycin, lincospectin (lincomycin/spectinomycin), neomycin, premafloxacin, spectinomycin, sulfamethoxazole/trimethoprim, and tetracycline against a total of 180 isolates of Actinobacillus pleuropneumoniae, Escherichia coli, and Salmonella choleraesuis (60 each) clinically isolated from pigs on farms in Taiwan from 1994 to 1996. No more than 3 isolates per farm were used. Ceftiofur had the highest activity in vitro against isolates of A. pleuropneumoniae, E. coli, and S. choleraesuis, with MIC90 values of 0.03, 2, and 1 microg/ml, respectively. Premafloxacin was highly active against isolates of A. pleuropneumoniae, E. coli, and S. choleraesuis, with MIC90 values of 2, 8, and 0.5 microg/ml, respectively, which were lower than those with enrofloxacin (MIC90 8, 32, and 2 microg/ml, respectively). Neomycin was moderately active against A. pleuropneumoniae and E. coli, with MIC90 values of 8 and 64 microg/ml, respectively, but was inactive with S. choleraesuis. Gentamicin showed high activity against A. pleuropneumoniae (MIC90 of 2 microg/ml) but was only moderately active with E. coli and S. choleraesuis (MIC90 of 64 and 32 microg/ml). Cephalothin was highly active against isolates of A. pleuropneumoniae (MIC90 of 1 microg/ml) but was inactive with E. coli (MIC90 of 128 microg/ml). Lincomycin had moderate activity (MIC90 of 32 microg/ml) against A. pleuropneumoniae. Chloramphenicol, lincomycin, and tetracycline were inactive with E. coli and S. choleraesuis (MIC90 > 128 microg/ml). In conclusion, ceftiofur and premafloxacin were highly active against isolates of A. pleuropneumoniae, E. coli, and S. choleraesuis, enrofloxacin and gentamicin were highly to moderately active; cephalothin was highly active against A. pleuropneumoniae and moderately active against S. cholearesuis; chloramphenicol, lincomycin, and tetracycline were active only with A. pleuropneumoniae; neomycin was moderately active against A. pleuropneumoniae and E. coli. The other antimicrobials tested were inactive. PMID- 11939338 TI - Placentitis, fetal pneumonia, and abortion due to Rhodococcus equi infection in a Thoroughbred. AB - Rhodococcus equi is a rare cause of equine abortion. This report describes pyogranulomatous placentitis and fetal pneumonia in a case of abortion from a Thoroughbred mare. Numerous Gram-positive coccobacilli were noted histologically within macrophages in placental and pulmonary lesions. Rhodococcus equi was isolated in pure culture from the placenta, lung, liver, kidney, and stomach content. This is the first description of placentitis due to Rhodococcus equi infection in a horse. PMID- 11939340 TI - Immunohistochemical demonstration of Francisella tularensis in lesions of cats with tularemia. AB - An immunohistochemical test was developed and validated for detection of Francisella tularensis antigen in tissues of cats with fatal tularemia. Ten cases of naturally occurring tularemia in cats were positive both by isolation of F. tularensis and immunohistochemical identification of F. tularensis antigen. Nine additional cases with lesions typical of tularemia were positive for F. tularensis antigen, although bacterial cultures were not performed. Immunohistochemical identification of F. tularensis in formalin-fixed tissue is valuable for establishing a rapid etiologic diagnosis under circumstances where fresh tissues may not be available for isolation and identification of the organism. PMID- 11939339 TI - Isolation of Arcanobacterium (Actinomyces) pyogenes from cases of feline otitis externa and canine cystitis. AB - Arcanobacterium pyogenes is a normal inhabitant of the mucous membranes of domestic animals, such as cattle, sheep, swine, and goats. It is also an opportunistic pathogen in these animals, where it causes a variety of purulent infections involving the skin, joints, and visceral organs. Two recent cases of isolation of A. pyogenes from companion animals are reported. In the first case, a cat presented with a chronic otitis externa, from which A. pyogenes was isolated in pure culture. The second case involved a dog with a urinary tract infection, where A. pyogenes was isolated from urine as the predominant bacterial species. In both cases, the A. pyogenes isolates were presumptively identified by macrobiochemical tests, and then their identities were confirmed by polymerase chain reaction analysis and 16S rRNA gene sequencing. PMID- 11939341 TI - Preliminary study on differentiation of Leptospira grippotyphosa and Leptospira sejroe from other common pathogenic leptospiral serovars in canine urine by polymerase chain reaction assay. AB - A multiplex polymerase chain reaction (PCR) method using primer sets of G1/G2 and B64-I/B64-II was validated to detect pathogenic leptospira serovars from canine urine samples. The PCR method was found to be specific and sensitive with a detection limit of 100 cells of Leptospira icterohaemorrhagiae per milliliter of urine. The primer set previously designated and erroneously transcribed B64-I/B64 II amplified a DNA fragment of 352 base pairs from Leptospira grippotyphosa and Leptospira sejroe but not from Leptospira autumnalis, Leptospira bratislava, Leptospira canicola, Leptospira hardjo, Leptospira icterohaemorrhagiae, and Leptospira pomona. From 100 diagnostic canine urine samples, 5 were found positive for Leptospira grippotyphosalsejroe with a PCR product of 352 base pairs and 6 were positive for other pathogenic leptospira serovars with a PCR product of 285 base pairs. One 285-base pair product was sequenced and found to be 99.3% homologous to the G1/G2 PCR fragment sequence reported previously. All 352-base pair PCR products of clinical samples and pure cultures of L. grippotyphosa and L. sejroe were sequenced. The 352-base pair fragment sequences of L. grippotyphosa and L sejroe were identical. Only 2 base pairs were found different between the sequences from pure cultures and those from clinical samples. Serum samples from 3 positive cases that generated a PCR product of 352 base pairs were tested by the microscopic agglutination test, and 2 were found to be positive for L. grippotyphosa (1:10,240 and 1:5,120), 1 was positive for L. grippotyphosa (1:320) or L. icterohaemorrhagiae (1:320). The results of this study suggest that the multiplex PCR with the primer set G1/G2 and the erroneously transcribed B64 I/B64-II may be able to differentiate L. grippotyphosa or L. sejroe from other pathogenic leptospira serovars commonly tested for in Canadian diagnostic laboratories. PMID- 11939342 TI - An immunoperoxidase monolayer assay for the detection of antibodies against swine influenza virus. AB - An immunoperoxidase monolayer assay (IPMA) has been developed to detect antibodies against swine influenza A virus (SIV) in pig sera. The test was evaluated by using sequential sera from pigs experimentally infected with H1N1 subtype of SIV. Two hundred field serum samples that had been examined by the hemagglutination inhibition (HI) test were also tested. Antibodies specific to SIV were detected as early as 3 days postinoculation (dpi) in the IPMA test as compared with 7 dpi by the HI test. Unlike HI, no serum treatment was required in the IPMA test. Regardless of the virus used in the test, IPMA detected antibodies to both H1N1 and H3N2 subtypes of SIV whereas HI detects antibodies against either H1N1 or H3N2, depending upon the virus used in the test. Results of this study indicate that IPMA is a useful test for screening of pig sera for SIV antibodies. PMID- 11939343 TI - Rapid detection of avian pneumovirus in tissue culture by microindirect immunofluorescence test. AB - An indirect immunofluorescence (IFA) test with a 96-well, flat-bottomed microplate was developed to detect avian pneumovirus (APV) antigen in Vero cell cultures. Samples of nasal turbinates and swabs from infraorbital sinuses and trachea were collected from 4-week-old poults experimentally inoculated with APV. The APV titers by tissue culture IFA staining were compared with that of visual reading of cytopathic effect (CPE). The ability of IFA staining to detect APV antigen correlated well with visualizing CPE. The use of IFA staining of Vero cell cultures allowed detection of APV in substantially less time than the use of visualizing CPE. In addition, the use of IFA allowed specific identification of the virus in cell culture. PMID- 11939344 TI - Neuroaxonal dystrophy in raccoons (Procyon lotor) from Iowa. AB - During a 12-month period (1998-1999), microscopic evidence of neuroaxonal dystrophy (NAD) in medullae oblongata of raccoons (Procyon lotor) was observed in 17/39 (47% prevalence in adults) from Iowa, USA. Three of the animals were kits (<3 months), 26 were between 1 and 2 years, and 10 were over 7 years. Lesions were not seen in the medullae of the 3 kits. In young adults, the lesions were mild and were seen in 7 animals. More severe lesions were present in the 10 older raccoons. Grossly, the brains were unremarkable. Microscopically, NAD was confined to the dorsal caudal medulla, where certain nuclei (predominantly gracilis and cuneate) were bilaterally affected. Severely affected animals had vacuolar degeneration of neurons or neuronal loss and extensive areas of spongiosis. Tests for the presence of PrP(res) in the brain were negative. Spongiotic areas often contained axonal spheroids. Degenerate neurons and axons occasionally contained amphophilic periodic acid-Schiff-positive granular material. There was a paucity of inflammatory cells in the affected areas. Since lesions were not present in kits, were either absent or mild in young adults, and were severe in older raccoons, the findings may be related to advancing age. Neuroaxonal dystrophy has not been previously reported in raccoons. Retrospective examination of raccoon brains from the eastern and northwestern areas of the country revealed very low prevalence of NAD. Because of the apparently high prevalence of this condition at this geographic location, factors other than age (genetic, nutritional, and/or environmental) may influence this degenerative process in the brains of raccoons in Iowa. PMID- 11939345 TI - The age of competence: an update on the international laboratory accreditation scene for veterinary testing laboratories. AB - Many changes have recently taken place in the world of laboratory accreditation. These changes include the increased use of voluntary standards in lieu of regulations, a move towards harmonization (equivalent results using voluntary standards) over standardization (using the same procedures or regulations), and an increased focus on competence, which includes the competence of laboratories to conduct testing, the competence of accreditation bodies to operate accreditation programs, the competence of bodies such as the National Cooperation for Laboratory Accreditation (NACLA) to recognize accreditation bodies as meeting the requirements of relevant standards, and the competence of organizations providing services to the accreditation process, such as the operation of proficiency testing programs. To describe these changes, a brief and general description of the International Laboratory Accreditation Cooperation accreditation scheme is provided, including an update on relevant decisions and activities in the United States and a description of the organization and activities of the newly formed NACLA. Following this discussion, with emphasis on veterinary testing, is an overview of several national and international organizations, including accreditation bodies, that promote harmonization, standardization, and analytical excellence. Also outlined are relevant activities of these organizations, an overview of some of the standards and guidelines they produce, and a description of how such organizations interact with each other and with laboratories seeking recognition for competence. Next is a brief discussion of recent developments and trends in laboratory accreditation, the impact of these developments, and the costs and benefits of accreditation to laboratories. Suggestions to veterinary laboratories for formulating strategy for keeping current with developments in accreditation and for determining quality goals are included. PMID- 11939346 TI - Field validation of a commercial blocking ELISA to differentiate antibody to transmissible gastroenteritis virus (TGEV) and porcine respiratory coronavirus and to identify TGEV-infected swine herds. AB - A commercially available blocking ELISA was analyzed for its ability to identify antibodies to porcine coronaviruses (transmissible gastroenteritis virus [TGEV] or porcine respiratory coronavirus [PRCV]), to differentiate antibodies to TGEV and PRCV, and to identify TGEV-infected herds. Nine sera from uninfected pigs, 34 sera from 16 pigs experimentally infected with TGEV, and sera from 10 pigs experimentally infected with PRCV were evaluated using both the TGEV/PRCV blocking ELISA and a virus neutralization (VN) assay. The ELISA was not consistently effective in identifying pigs experimentally infected with TGEV until 21 days postinfection. Sera from 100 commercial swine herds (1,783 sera; median 15 per herd) were similarly evaluated using both tests. Thirty of these commercial herds had a clinical history of TGEV infection and a positive TGEV fluorescent antibody test recorded at necropsy within the last 35 months, while 70 herds had no history of clinical TGEV infection. The blocking ELISA and the VN showed good agreement (kappa 0.84) for the detection of porcine coronavirus antibody (TGEV or PRCV). The sensitivity (0.933) of the ELISA to identify TGEV infected herds was good when considered on a herd basis. The ELISA was also highly specific (0.943) for the detection of TGEV-infected herds when the test results were evaluated on a herd basis. When sera from specific age groups were compared, the ELISA identified a greater proportion (0.83) of pigs in herds with TGEV antibody when suckling piglets were used. In repeatability experiments, the ELISA gave consistent results when the same sera were evaluated on different days (kappa 0.889) and when sera were evaluated before and after heating (kappa 0.888). The blocking ELISA was determined to be useful for herd monitoring programs and could be used alone without parallel use of the VN assay for the assessment of large swine populations for the detection of TGEV-infected herds. PMID- 11939347 TI - Recent progress on the molecular organization of myelinated axons. AB - The structure of myelinated axons was well described 100 years ago by Ramon y Cajal, and now their molecular organization is being revealed. The basal lamina of myelinating Schwann cells contains laminin-2, and their abaxonal/outer membrane contains two laminin-2 receptors, alpha6beta4 integrin and dystroglycan. Dystroglycan binds utrophin, a short dystrophin isoform (Dp116), and dystroglycan related protein 2 (DRP2), all of which are part of a macromolecular complex. Utrophin is linked to the actin cytoskeleton, and DRP2 binds to periaxin, a PDZ domain protein associated with the cell membrane. Non-compact myelin--found at incisures and paranodes--contains adherens junctions, tight junctions, and gap junctions. Nodal microvilli contain F-actin, ERM proteins, and cell adhesion molecules that may govern the clustering of voltage-gated Na+ channels in the nodal axolemma. Na(v)1.6 is the predominant voltage-gated Na+ channel in mature nerves, and is linked to the spectrin cytoskeleton by ankyrinG. The paranodal glial loops contain neurofascin 155, which likely interacts with heterodimers composed of contactin and Caspr/paranodin to form septate-like junctions. The juxtaparanodal axonal membrane contains the potassium channels Kv1.1 and Kv1.2, their associated beta2 subunit, as well as Caspr2. Kv1.1, Kv1.2, and Caspr2 all have PDZ binding sites and likely interact with the same PDZ binding protein. Like Caspr, Caspr2 has a band 4.1 binding domain, and both Caspr and Caspr2 probably bind to the band 4.1 B isoform that is specifically found associated with the paranodal and juxtaparanodal axolemma. When the paranode is disrupted by mutations (in cgt-, contactin-, and Caspr-null mice), the localization of these paranodal and juxtaparanodal proteins is altered: Kv1.1, Kv1.2, and Caspr2 are juxtaposed to the nodal axolemma, and this reorganization is associated with altered conduction of myelinated fibers. Understanding how axon-Schwann interactions create the molecular architecture of myelinated axons is fundamental and almost certainly involved in the pathogenesis of peripheral neuropathies. PMID- 11939348 TI - Degeneration and regeneration of the peripheral nervous system: from Augustus Waller's observations to neuroinflammation. AB - This review article on the degeneration and regeneration of peripheral nerve fibers was presented as a Plenary Lecture at the 2001 meeting of the Peripheral Nerve Society. It is accompanied by a reprint of Augustus Waller's 1850 article, which gave rise to the pathologic process termed Wallerian degeneration. This review is focused on the role of neuroinflammation in Wallerian degeneration and how immune mediators contribute to both axonal degeneration and regeneration. Similarities and differences between the PNS and CNS in terms of inflammation and microglial activation after nerve injury are discussed, and point towards progress in understanding the failure of nerve fiber regeneration in the CNS. PMID- 11939349 TI - Endoneurial remodeling by TNFalph- and TNFalpha-releasing proteases. A spatial and temporal co-localization study in painful neuropathy. AB - Peripheral nerve injury causing Wallerian degeneration results in endoneurial remodeling initiated by an increase in tumor necrosis factor-alpha (TNF), which is activated from its precursor by extracellular proteases of the matrix metalloproteinase (MMP) family. We used immunohistochemistry to analyze the distribution of TNF, TNF-releasing MMPs, including gelatinases (MMP-2 and MMP-9), and TNF-alpha converting enzyme (TACE) in painful neuropathy caused by chronic constriction injury of rat sciatic nerve. Tissue was analyzed at the injury site and in the corresponding L4 and L5 dorsal root ganglia (DRG) throughout the time course of the neuropathy. Using confocal laser scanning microscopy, we co localized TNF with each MMP, and observed spatial and temporal distinction in their distribution. TNF co-localized in vessel endothelium with MMP-2 and in macrophages with MMP-9 and TACE at the period of active immune cell migration. TNF co-localized with myelin degrading MMP-9 within Schwann cells during demyelination, and intraaxonally during remyelination. These studies were performed to explore the role of basal-lamina degrading gelatinases and other TNF releasing MMPs in TNF-mediated Wallerian degeneration. The data provided in this study may be useful in designing selective therapy for painful neuropathy using synthetic hydroxamate MMP inhibitors. PMID- 11939350 TI - Aggravated reperfusion injury in STZ-diabetic nerve. AB - The streptozocin (STZ)-diabetic nerve manifests increased morphological susceptibility to a superimposed acute ischemic injury, and reperfusion injury exaggerates ischemic nerve pathology. To determine whether STZ-diabetic nerves are susceptible to reperfusion, we evaluated the pathological consequences after 2.5 hours of ischemia followed by 3 and 24 hours of reperfusion in a 20-week STZ diabetic rat sciatic nerve. After 3 hours of reperfusion, endoneurial edema developed in diabetic nerves, whereas non-diabetic controls showed mild or no edema. Morphometric analysis of endoneurial edema, quantified by the total transverse fascicular area and the point-count score of endoneurial structureless space, confirmed significantly more reperfusion-induced edema at thigh and knee levels in diabetic nerves than in controls. Reperfusion caused a significant increase in the number of endoneurial mast cells at the thigh level in diabetic nerves. After 24 hours of reperfusion, there were striking morphological anomalies of myelinated nerve fibers in diabetic nerves, without any observable changes in control nerves. In conclusion, we have demonstrated that STZ-diabetes exacerbates the morphological change to reperfusion. Diabetes therefore renders the microvasculature more vulnerable to the deleterious effects of ischemia/reperfusion. PMID- 11939351 TI - Expression and localization of insulin receptor in rat dorsal root ganglion and spinal cord. AB - The expression and localization of the insulin receptor (IR) was examined in rat dorsal root ganglia (DRG) and spinal cord using Western blotting, in situ hybridization and immunocytochemistry. Western blotting showed that the molecular weight of the IR beta subunit was higher in PNS than that found in CNS. Both IR mRNA and protein expressions were highest in small-sized sensory DRG neurons and myelinated sensory root fibers expressed higher levels of IR protein than myelinated anterior root fibers. In the spinal cord, IR immunoreactive neurons were present in lateral lamina V and in lamina X, suggesting the presence of IR in nociceptive pathways. Electronmicroscopy of DRGs revealed a polarized localization of the IR in abaxonal Schwann cell membranes, outer mesaxons in close vicinity to tight junctions of both myelinating and non-myelinating Schwann cells and to plasma membranes of sensory neurons. From these findings, we speculate that insulin may play a role in sensory fibers involved in nociceptive function often perturbed in diabetic neuropathy. The high expression of IR localizing to tight junctions of dorsal root mesaxons of DRGs may suggest a regulatory role on barrier functions compensating for the lack of a blood-nerve barrier in dorsal root ganglia. This is consistent with the colocalization of IR with tight junctions of the paranodal barrier and endoneurial endothelial cells in peripheral nerve. PMID- 11939352 TI - Anti-LM1 antibodies in the sera of patients with Guillain-Barre syndrome, Miller Fisher syndrome, and motor neuron disease. AB - This study is designed to establish whether sialosylneolactotetraosylceramide (LM1), a major component of human peripheral nerve ganglioside, is a potential target antigen for the development of peripheral autoimmune neuropathies such as Guillain-Barre syndrome (GBS) and Miller Fisher syndrome (MFS). Serum antibodies against LM1 in 116 patients with GBS, 56 patients with MFS, 88 patients with motor neuron disease (MND) and 60 normal control subjects were quantified using enzyme-linked immunosorbent assay (ELISA). The presence of anti-LM1 antibodies were confirmed using an immunostaining method on high-performance thin-layer chromatographic plates (HPTLC). Anti-LM1 IgG antibodies were detected in 22% (25/116) of patients with GBS. The ratio of the demyelination type to the axonal type of GBS was approximately 3:1. Among the 25 anti-LM1-positive GBS patients, additional anti-GM1 IgG antibodies were detected in 7 patients, 4 of whom possessed the axonal form of GBS. Anti-LM1 antibodies were also detected in a significant portion of patients with MFS (20%, 11/56). In contrast, anti-LM1 antibodies were detected in only 2% (2/88) of patients with MND, and 7% (4/60) of normal control subjects. The results of this study suggest that serum antibodies against LM1 may have a pathogenic role in the development of GBS and MFS. PMID- 11939353 TI - Cryoglobulinemia is a frequent cause of peripheral neuropathy in undiagnosed referral patients. AB - Cryoglobulinemia represents an emerging cause of peripheral neuropathy, especially in Southern Europe, in view of its relationship with hepatitis C virus infection. In a series of 100 consecutive referral patients with uncharacterized peripheral neuropathies, we systematically investigated cryoglobulinemia to assess its diagnostic yield. The most frequent diagnosis was hereditary neuropathy (33%), 29% were acquired neuropathies of different types, and no cause could be identified in 27%. Cryoglobulinemic neuropathy was diagnosed in 11 patients (7 women and 4 men), aged 54-77 (mean = 63.5 years), most presenting with sensory polyneuropathy, often asymmetrical. Cryoglobulin was also detected in 2 additional patients in whom a final diagnosis of non-Hodgkin lymphoma was made. Purpura was absent in 4 patients (and in 2 with lymphoma), or restricted to discrete manifestations in the remaining patients, which did not provide a clue to the diagnosis. Thus, search for cryoglobulin proves useful in a substantial number of undiagnosed peripheral neuropathies (11% to 13% in our series), even in the absence of typical skin lesions, and it is recommended as a first-line investigation in patients with unexplained neuropathy presenting in middle to older age. PMID- 11939354 TI - Vehicle action: effective policy for controlling drunk and other high-risk drivers? AB - License suspension effectively reduces recidivism and crash involvement of those convicted of driving while impaired (DWI). The impact of this sanction, however, is being reduced by the large number of offenders (up to 75%) who drive even though suspended. To deal with this problem. several states have enacted laws providing for vehicle impoundment, immobilization, or forfeiture for repeat DWI offenders and for driving while suspended (DWS) offenders. Although a 1992 review of vehicle sanctions for DWI and DWS offenders showed 32 states with such laws, they were infrequently applied. Further, none of those laws had been adequately evaluated. This paper reviews the studies of vehicle action programs in California, Minnesota, New York, Ohio, Oregon, and Washington that have been applied broadly enough to permit evaluation. Although none of the studies has applied random assignment to ensure equal groups. several have applied sufficient statistical controls to provide reasonably credible results. All the programs reviewed showed positive effects. including some relatively large recidivism reductions, from denying offenders the use of their vehicles for 1-6 months. Highlighted in this review are several issues that appear to be important to the effectiveness of vehicle sanctioning programs. PMID- 11939355 TI - An exploratory study of the relationship between road rage and crash experience in a representative sample of US drivers. AB - The phenomenon of road rage has been frequently discussed but infrequently examined. Using a representative sample of 1382 US adult drivers, who were interviewed in a 1998 telephone survey, exploratory analyses examined the relationship between self-reported measures of road rage, generally hazardous driving behaviors, and crash experience. Regarding specific road rage behaviors, most respondents reported having engaged in verbal expressions of annoyance; however only 2.45% reported ever having been involved in direct confrontation with another car or driver. After controlling for gender, age. driving frequency, annual miles driven and verbal expression, an angry/threatening driving subscale of road rage was significantly associated with hazardous driving behaviors that included frequency of driving over the legal blood alcohol limit, receipt of tickets in the past year. and habitually exceeding the speed limit as well as crash experience. However, the verbal/frustration expression subscale was not associated with crash experience or hazardous driving indicators, except for number of tickets, after controlling for other crash-related factors such as gender and age. Direct confrontation by deliberately hitting another car or leaving the car to argue with and/or injure another driver was rarely reported. Results suggest that angry/threatening driving is related to crash involvement; however, after controlling for exposure and angry/threatening and hazardous driving the relationship of milder expressions of frustration while driving and crash involvement was not significant. PMID- 11939356 TI - Characteristics of traffic crashes in Maryland (1996-1998): differences among the youngest drivers. AB - Motor vehicle crashes (MVCs) are the leading cause of death among teenagers in the US. The present study examines how crash rates and crash characteristics differed among drivers aged 16-21 in the state of Maryland from 1996 to 1998. The results show that, based on police reports. the youngest drivers have the highest rate of MVCs per licensed driver and per annual miles driven. Furthermore, crash characteristics suggest that inexperience rather than risky driving may account for the differing rates. Drivers closer to the age of 16 had their crashes under the safest conditions: during the day in clear weather while drinking less. PMID- 11939357 TI - A comparison of safety belt use between commercial and noncommercial light vehicle occupants. AB - The purpose of this study was to conduct an observational survey of safety belt use to determine the use rate of commercial versus noncommercial light-vehicle occupants. Observations were conducted on front-outboard vehicle occupants in eligible commercial and noncommercial vehicles in Michigan (i.e.. passenger cars, vans/minivans, sport-utility vehicles, and pickup trucks). Commercial vehicles that did not fit into one of the four vehicle type categories, such as tractor trailers, buses, or heavy trucks, were not included in the survey. The study found that the restraint use rate for commercial light-vehicle occupants was 55.8% statewide. The statewide safety belt use rate for commercial light-vehicles was significantly lower than the rate of 71.2% for noncommercial light-vehicles. The safety belt use rate for commercial vehicles was also significantly different as a function of region, vehicle type, seating position, age group, and road type. The results provide important preliminary data about safety belt use in commercial versus noncommercial light-vehicles and indicate that further effort is needed to promote safety belt use in the commercial light-vehicle occupant population. The study also suggests that additional research is required in order to develop effective programs that address low safety belt use in the commercial light-vehicle occupant population. PMID- 11939358 TI - Toll road crashes of commercial and passenger motor vehicles. AB - Revenue-collection data from toll roads allow for accurate estimates of miles driven by vehicle type and, when combined with crash data, valid estimates of crash involvements per mile driven. Data on vehicle-miles traveled and collisions were obtained from toll road authorities in Florida. Kansas, and New York. In addition, state crash files and published vehicle-miles of travel were obtained for toll roads in Illinois. Indiana, Ohio, and Pennsylvania. Large commercial motor vehicles were significantly underinvolved in single-vehicle crashes on all state toll roads. In five states, commercial motor vehicles were significantly overinvolved in multiple-vehicle crashes relative to passenger vehicles; the exceptions were Kansas, where they had significantly lower multiple-vehicle involvement rates, and Indiana. where there were no significant differences in multiple-vehicle involvements by vehicle type. The risk of commercial motor vehicle involvement in multiple-vehicle crashes resulting in deaths or serious injuries was double that of passenger vehicles in the two states (Ohio and Pennsylvania) that identified serious injuries. Whether crash rates, on toll roads of commercial motor vehicles are higher or lower than those of passenger vehicles appears to depend on the type of crash, specific toll road. and traffic density. PMID- 11939359 TI - Sobriety tests for low blood alcohol concentrations. AB - The legal limits of blood alcohol concentration (BAC) for operators of commercial vehicles throughout the United States, and underage drivers in most states, are approximately half the 0.08-0.10% imposed on other drivers. Easily administered measures involving performance and appearance are needed to establish probable cause for requesting breath tests. Several measures showing relationships to blood alcohol were examined for their ability to distinguish BACs under and over 0.04% within the 0.00-0.08% range. Measures of heat loss. pupillary response, balance, complex tasks, and accuracy under speed were studied in controlled experiments with alcohol-dosed subjects. The only reliable index of blood alcohol was horizontal gaze nystagmus (HGN) as administered in the standardized field sobriety test (SFST), using alternative scoring criteria. Although other measures evidenced intra-individual change at low BACs, the magnitude of change was greatly exceeded by inter-individual differences, thwarting their use in detecting drivers with BACs lower than 0.08%. HGN, however, proved as valid in detecting BACs in the 0.04-0.08% range as at the higher levels of the SFST. It is also as valid when administered to a seated subject as one standing, making it of particular benefit in enforcement, where the behavior of low BAC drivers rarely provides probable cause to request that they leave a vehicle to be tested in a standing position. PMID- 11939360 TI - Driver injury severity: an application of ordered probit models. AB - This paper describes the use of ordered probit models to examine the risk of different injury levels sustained under all crash types, two-vehicle crashes, and single-vehicle crashes. The results suggest that pickups and sport utility vehicles are less safe than passenger cars under single-vehicle crash conditions. In two-vehicle crashes, however, these vehicle types are associated with less severe injuries for their drivers and more severe injuries for occupants of their collision partners. Other conclusions also are presented; for example. the results indicate that males and younger drivers in newer vehicles at lower speeds sustain less severe injuries. PMID- 11939361 TI - International comparison of costs of a fatal casualty of road accidents in 1990 and 1999. AB - The purpose of this study is to assemble information of costs per fatal casualty in traffic accidents, adopted by authorities in different countries. analyse and compare these figures as well as the methods used for estimating these values. A questionnaire was sent to 19 countries from which 11 gave information on cost per fatality and methods of valuation. The costs per fatality, usually defined as direct and indirect costs plus a value of safety per se, are compared both between countries and over time, 1990 and 1999, for each country. The average cost per fatality has increased between 1990 and 1999 (fixed prices) due to both changes in the methodology and changes of valuations. Great Britain, New Zealand, Sweden and the US conduct own willingness-to-pay (WTP) surveys, while the Netherlands and Norway make reviews of these studies. In Finland, the cost per fatality is a combination of the value of lost productivity and the cost of care for an institutionalised disabled person. In Australia, Austria, Germany and Switzerland, the cost per fatality is estimated as a value of lost productivity and an addition of a human cost based on compensation payments or insurance payments. Estimates from recently conducted WTP surveys or meta-analyses are considered in Austria, Finland and Sweden, however, not yet adopted as official values for use in road traffic planning. PMID- 11939362 TI - The antilock braking system anomaly: a drinking driver problem? AB - Antilock braking systems (ABS) have held promise for reducing the incidence of accidents because they reduce stopping times on slippery surfaces and allow drivers to maintain steering control during emergency braking. Farmer et al. (Accident Anal. Prevent. 29 (1997) 745) provide evidence that antilock brakes are beneficial to nonoccupants: a set of 1992 model General Motors vehicles equipped with antilock brakes were involved in significantly fewer fatal crashes in which occupants of other vehicles, pedestrians, or bicyclists were killed. But, perversely, the risk of death for occupants of vehicles equipped with antilock brakes increased significantly after adoption. Farmer (Accident Anal. Prevent. 33 (2001) 361) updates the analysis for 1996- 1998 and finds a significant attenuation in the ABS anomaly. Researchers have put forward two hypotheses to explain this antilock brake anomaly: risk compensation and improper operation of antilock brake-equipped vehicles. We provide strong evidence for the improper operation hypothesis by showing that the antilock brake anomaly is confined largely to drinking drivers. Further, we show that the attenuation phenomenon occurs consistently after the first three to four years of vehicle service. PMID- 11939363 TI - Is the claim that 'variance kills' an ecological fallacy? AB - Over the past 15 years, a number of studies have reported positive correlations between estimated traffic crash rate and the dispersion of vehicle speeds. These correlations have on occasion been interpreted as supporting the view that slower and/or faster drivers have higher crash risks, or that speed variance itself is a causal factor for individual crash risk. This paper points out first that such positive correlations can be expected in situations where individual crash risk is either an increasing, or a decreasing, or a U-shaped function of speed, and so the correlations in themselves provide no evidence concerning the relation between speed and crash risk for individuals. Second, since such correlations can be expected in circumstances where individual risk is independent of speed variance, observation of these correlations provides no support for the hypothesis that increases in speed variance increase individual risk. PMID- 11939364 TI - The effectiveness of antilock braking systems in reducing accidents in Great Britain. AB - Antilock braking systems (ABS) are fitted to many new cars with the aims of improving their ability to steer while braking heavily and of reducing stopping distances on some road surfaces. This paper presents the findings of a project that assessed the effectiveness of ABS in reducing accidents in Great Britain. A large postal survey was carried out of the owners of modern cars. asking for details of any accidents in which they had been involved during the previous year as well as factors that might influence their likelihood of being involved in an accident. Questions were also asked to test respondents' knowledge of ABS. ABS cars ditfered in several respects from non-ABS cars, as did the two groups of drivers. Consequently, a sophisticated statistical analysis was required to provide unbiased estimates of the effectiveness of ABS. Various results did not achieve statistical significance, but the overall stability of the results indicates that real effects have been measured. They confirm that ABS does have the potential to reduce the number of accidents, but show that this has not been fully achieved. One reason may be that many drivers have little or no knowledge of ABS. PMID- 11939365 TI - Effects of road geometry and traffic volumes on rural roadway accident rates. AB - This paper revisits the question of the relationship between rural road geometric characteristics, accident rates and their prediction, using a rigorous non parametric statistical methodology known as hierarchical tree-based regression. The goal of this paper is twofold: first, it develops a methodology that quantitatively assesses the effects of various highway geometric characteristics on accident rates and, second, it provides a straightforward, yet fundamentally and mathematically sound way of predicting accident rates on rural roads. The results show that although the importance of isolated variables differs between two-lane and multilane roads, 'geometric design' variables and pavement condition' variables are the two most important factors affecting accident rates. Further, the methodology used in this paper allows for the explicit prediction of accident rates for given highway sections, as soon as the profile of a road section is given. PMID- 11939366 TI - Multidisciplinary in-depth investigations of head-on and left-turn road collisions. AB - To enhance traffic safety, a multidisciplinary Road Accident Investigation Team was established in Denmark for a 2-year trial period. The objective was to conduct in-depth investigations of specific types of accidents, and to identify effective preventive measures. The team consisted of a road engineer, a vehicle inspector, a police superintendent, a psychologist and a physician. Seventeen serious head-on collisions as well as 17 left-turn collisions were analysed. In collecting data, police reports were supplemented by the team's investigation of accident sites and vehicles involved, and interviews were carried out with the involved road users and witnesses. The drivers, to whom the accident factors were primarily related in the head-on collisions, were characterised by their conscious risk-taking behaviour. They were all males; several of them were under age 40 and had earlier traffic and/or drug convictions. The main accident factors were excessive speed, drunk driving and driving under the influence of illegal drugs. In the left-turn accidents, the most common accident factors were attention errors, and it was also noted that elderly drivers ( > 74) were over represented. The synergy effect of working as a multidisciplinary team proved fruitful. It resulted in a more precise knowledge of the road accident circumstances and of contributing factors leading up to the accidents. Due to the great demand on resources, only a limited number of accidents could be analysed, but the results provide a basis for further and more targeted research. PMID- 11939367 TI - Self-reports of aberrant behaviour on the roads: errors and violations in a sample of Greek drivers. AB - Recent studies have shown that unsafe driver acts can be classified into two distinct categories (i.e.. errors and violations) entailing different measures for reducing road traffic accidents. A survey of over 1,400 drivers in Greece is reported in which a variety of aberrant driving behaviours have been identified. The present study has confirmed the results of earlier studies in the driving populations of Britain. Australia and Sweden that errors and violations are the major determinants in the factor structure of aberrant behaviours. Three types of violations were identified including, highway code-, aggressive- and parking violations (or situational violations). Mistakes and lapses were two major forms of errors. The factor analysis identified another two classes of behaviour that could not be accounted for very well by the error-violation distinction. One class of behaviours referred to a state of low preparedness and negligence while the other class referred to communication errors and 'social disregard' for the other road users. Each class of behaviour was found to have different demographic correlates. Finally, accident liability was predicted by self-reported tendency to commit highway-code violations, but not by tendency to make errors due to failures of judgement (i.e.. mistakes) or lapses. Aggressive violations were significantly related to involvement in speeding convictions and law-breaking whilst highway-code violations were related to speeding convictions only. PMID- 11939368 TI - A note on the variance of paired comparisons estimates. AB - The method of paired comparisons to estimate treatment effectiveness was introduced by Evans (Evans L. Double pair comparison--a new method to determine how occupant characteristics affect fatality risk in traffic crashes. Accident Analysis and Prevention 1985;12:217-27). It is similar in form to other effectiveness estimates based on odds ratios using independent groups. Therefore, it has been generally assumed that the variance is computed in the same way. In this note, it is demonstrated. using a simple binomial model and linear approximation, that the variance is lower for paired comparisons estimates than for odds ratios estimates based on independent groups. In order to use odds ratios, there must be a treated group and an untreated group. Within each group there are occurrences of an event against which treatment effectiveness is being estimated and, also, occurrences of different event that is considered unamenable to the treatment. The treatment effectiveness, e, is estimated by 1-R where R is the ratio of amenable type events to unamenable ones in the treated group, divided by the same ratio for the untreated group. A distinction is made between 'real' paired comparisons and odds ratios based on independent data. An example of the independent case is. x is the number of fatalities in frontal crashes without air bags; y the number of fatalities in non-frontal crashes without air bags; s the number of fatalities in frontal crashes with air bags, and t the number of fatalities in non-frontal crashes with air bags. While fatalities in non-frontal crashes serve as denominators in R, a particular frontal crash is not paired with one particular non-frontal crash. In this case, in which all the data are independent, the variance of e is approximately R2(1/x + 1/y + 1/s + 1/t), a result which is consistent with well known results about the log odds ratio. For an example of real paired comparisons, we consider fatalities in cars that have a driver and exactly one unbelted right front seat passenger. Suppose there were x driver fatalities and y passenger fatalities in the cars in which the driver was also unbelted and s driver fatalities and t passenger fatalities in the cars in which the driver was belted. Since the fate of the passenger would not be amenable to 'treating' the driver, the same estimate of belt effectiveness based on these data. 1 - (s/t)/(x/y), is reasonable. In this case, x and y, and s and t are not independent. This is due to the fact that while the overall probability of fatality in a crash is very low, the conditional probability of fatality given that someone else in the car died is greater than the unconditional probability of fatality. Under these circumstances, the variance of the paired comparisons estimate is reduced. Published by Elsevier Science Ltd. PMID- 11939369 TI - Temperature-induced formation of a non-native intermediate state of the all beta sheet protein CD2. AB - Domain 1 of the cell adhesion protein CD2 (CD2-1) has an all beta-structure typical of proteins belonging to the immunoglobin superfamily. It has a remarkable ability to fold as a native monomer or a metastable intertwined dimer. To understand the origin of structural rearrangements of CD2-1, we have studied equilibrium unfolding of the protein using various biophysical spectroscopic techniques. At temperatures above approx 68 degrees C, a partially folded state of CD2-1 (H state) with a distinct secondary structure, involving largely exposed aromatic and hydrophobic residues and a substantially perturbed tertiary structure, is observed. In contrast, an unfolded state (D state) of CD2-1 with random-coil-like secondary and tertiary structures is observed in 6 M GuHCl. This partially folded high-temperature state has increased negative molar ellipticity at 222 nm in far-ultraviolet CD spectra, implying formation of a non-native helical conformation. The existence of this non-native high-temperature intermediate is consistent with relatively high intrinsic helical propensities in the primary sequence of CD2-1. This conformational flexibility may be important in the observed domain swapping of CD2-1. PMID- 11939370 TI - A pH dependence study on the unfolding and refolding of apoazurin: comparison with Zn(II) azurin. AB - Azurin, a small blue copper protein from the bacterial species Pseudomonas aeruginosa, is mostly a beta-sheet protein arranged into a single domain. Previous folding studies have shown that the equilibrium denaturation of the holoprotein follows a two-state process; however, upon removal of the copper, the denaturation had been reported to follow a three-state process. The two unfolding transitions measured for apoazurin had been thought to arise from two different folding domains. However, in the present work, we found that the denaturation of apoazurin occurs over a single transition and we determined the folding free energy to be -27.8 +/- 2.4 kJ mol(-1). From this measurement along with measurements previously reported for the unfolding of the holoazurin, we were able to determine that Cu(II) and Cu(I) stabilize the native structure by 25.1 +/ 6.9 kJ/mol and 12.9 +/- 8.1 kJ/mol, respectively. It is our contention that the second transition displayed in the denaturation curves previously reported for apoazurin arise from protein heterogeneity-in particular, from the presence of Zn(II) azurin. We extended our investigation into the denaturation of Zn(II) azurin at pH 6.0 and 7.5. The equilibrium denaturation studies show that the zinc ion significantly stabilizes the native-state structure at pH 7.5 and very little at the lower pH. We attribute the decrease in the stabilizing effect of the zinc ion with decreasing pH to the protonation of two histidinyl side chains. When protonated the ligands, His 46 and His 117, are incapable of binding a metal ion. Further, comparing the denaturation curves of Zn(II) azurin measured by circular dichroism with those measured by fluorescence indicates that the denaturation of Zn(II) azurin is far less simple than the denaturation of apoazurin. PMID- 11939371 TI - Regulation of ion channels by integrins. AB - Ion channels are regulated by protein phosphorylation and dephosphorylation of serine, threonine, and tyrosine residues. Evidence for regulation of channels by tyrosine phosphorylation comes primarily from investigations of the effects of growth factors, which act through receptor tyrosine kinases. The purpose of the present work is to summarize evidence for the regulation of ion channels by integrins, through their downstream, nonreceptor tyrosine kinases. We review both direct and indirect evidence for this regulation, with particular emphasis on Ca2+-activated K+ and voltage-gated Ca2+ channels. We then discuss the critical roles that cytoskeletal, focal-adhesion, and channel-associated scaffolding proteins may play in localizing nonreceptor tyrosine kinases to the vicinity of ion channels. We conclude by speculating on the physiological significance of these regulatory pathways. PMID- 11939372 TI - Review of nonlinear analysis of proteins through recurrence quantification. AB - This review considers the use of a nonlinear signal analysis tool, recurrence quantification analysis, as a method to study sequence/structure relationships of proteins. Four broad categories are discussed: (1) a point of view involving information contained in deterministic aspects of hydrophobicity; (2) the analysis of protein hydrophobicity "singularities"; (3) time-series analysis of protein dynamics simulations; and (4) prediction of protein secondary structure. PMID- 11939374 TI - Leading and managing public health networks. PMID- 11939375 TI - Philosophy of public health: lessons from its history in England. AB - In a modern industrialized society it was essential for humankind to be protected by a strong medical presence, to establish a uniform system for public health everywhere in that society, to make public health independent from the welfare system, and to harmonize the relationship between personal hygienic rules and local authority rules. The society in Japan has developed on the basis of those four philosophical principles of public health, and has enabled the people to have the longest life expectancy in the world. However, the public health system in this society is now in a critical situation resulting from the long life expectancy. How can we rebuild the role of public health in this country? A society with a long life expectancy has to face a wide variety of health conditions. However, the health insurance system in Japan does not generally provide any service for a patient without specific symptoms. Consequently, to help people become aware of their own health condition, comprehensive public services such as general health check-ups, screening for cancers, and health counselling have been established locally by all 3250 municipal authorities. In promoting those services, people must learn how to draw the boundary-line between 'public' and 'private' in affairs of health to understand the importance of 'hygienic rules based on counsels for personal self-government', as Simon suggested 100 years ago, and health professionals must develop a public health system which depends not only on regulatory laws but also on scientific findings, as Rumsey advocated 150 years ago. PMID- 11939373 TI - Pore-forming protein structure analysis in membranes using multiple independent fluorescence techniques. AB - A large number of transmembrane proteins form aqueous pores or channels in the phospholipid bilayer, but the structural bases of pore formation and assembly have been determined experimentally for only a few of the proteins and protein complexes. The polypeptide segments that form the transmembrane pore and the secondary structure that creates the aqueous-lipid interface can be identified using multiple independent fluorescence techniques (MIFT). The information obtained from several different, but complementary, fluorescence analyses, including measurements of emission intensity, fluorescence lifetime, accessibility to aqueous and to lipophilic quenching agents, and fluorescence resonance energy transfer (FRET) can be combined to characterize the nature of the protein-membrane interaction directly and unambiguously. The assembly pathway can also be determined by measuring the kinetics of the spectral changes that occur upon pore formation. The MIFT approach therefore allows one to obtain structural information that cannot be obtained easily using alternative techniques such as crystallography. This review briefly outlines how MIFT can reveal the identity, location, conformation, and topography of the polypeptide sequences that interact with the membrane. PMID- 11939376 TI - Public health practitioners in NHS hospital trusts: the impact of 'medical care epidemiologists'. AB - BACKGROUND: In the late 1990s, one NHS region created posts of 'medical care epidemiologists' (MCEs) to employ public health skills in NHS Trusts in pursuit of the 'evidence-based medicine' agenda. METHODS: A qualitative interview study and documentary analysis of MCEs' activities were carried out, followed by case studies of selected activities with plausibility of claims assessed by a 'counterfactual panel'. RESULTS: Most MCEs were centrally involved in the following: Trust committee structures related to clinical effectiveness; service review and evaluation; development of information systems or of access to existing systems to support clinical effectiveness; training in critical appraisal or similar skills; local development of clinical guidelines. A minority adopted narrower roles; only one had concentrated on identifying usable clinical outcome measures, an original objective of the MCE initiative. Case studies of selected activities showed the following: the government's clinical governance agenda created a receptive context in which Trusts connected their MCE's appointment with an external policy agenda; the activities undertaken were nevertheless chosen and shaped by the MCEs themselves, leading to a close fit between the activity and their individual skills and interests; the main impact of MCE activity resulted from deployment of these in ways that transmitted messages about changing organizational culture, rather than from the formal output of their activities. CONCLUSIONS: It is unrealistic to expect individuals to change the culture of whole organizations. In the right policy context, a real contribution can be made by individuals with appropriate skills and strong personal agendas consonant with, but not determined by organizational agendas. PMID- 11939377 TI - NICE and LICE: lessons from a health district. National Institute for Clinical Excellence. Local institutes of clinical excellence. AB - This paper describes experiences of implementing guidance from the National Institute for Clinical Excellence (NICE) in the East Riding and Hull Health Authority area. Attempts have been made to integrate NICE guidance with the decision-making, service planning and monitoring processes but overall progress has been limited. Contributory reasons are explored, along with suggestions on how to ensure that NICE can help ensure effective and equitable services. PMID- 11939379 TI - Injury morbidity in 18-64-year-olds: impact and risk factors. AB - BACKGROUND: Non-fatal injury is an important public health problem but is thought to be difficult to quantify. This study aimed to estimate the extent of disability attributable to injury in the working age population, and its impact on quality of life, as well as identifying factors associated with an increased risk of disabling and non-disabling injury. METHODS: Secondary analysis was carried out of data obtained from a postal questionnaire survey of 8889 18-64 year-olds randomly selected from computerized general practitioner records in Oxfordshire, Buckinghamshire, Berkshire and Northamptonshire. RESULTS: Sixteen per cent of survey respondents reported an injury requiring medical attention in the previous 12 months, 5 per cent reported an injury that had disabled them for more than 1 month, and a further 5 per cent a longstanding disability as a result of injury. The point prevalence of disability as a result of injury was estimated to be 6.4 per cent. SF-36 scores suggest that the quality of life of people reporting injury-related disability was markedly reduced. Social class is associated less with injury morbidity than with injury mortality. Sport was the commonest cause of all injuries, and of disabling injuries. There was a dose response relationship between vigorous exercise and injury. CONCLUSIONS: Injury is a significant cause of disability in the working age population. It is potentially feasible to monitor injury-related disability in postal questionnaire surveys. Estimates of health gain to be achieved from participation in sport should take account of injury-related disability. PMID- 11939378 TI - Variation in coronary artery bypass grafting, angioplasty, cataract surgery, and hip replacement rates among primary care groups in London: association with population and practice characteristics. AB - BACKGROUND: Operation rates for common surgical procedures are known to vary widely both between areas within the same country and between countries. However, no studies in the United Kingdom have yet examined the variation in the use of important groups of surgical procedures among primary care groups. METHODS: Data were obtained from the NHS Executive and the Department of Health on each of the 66 primary care groups in London that existed on 1 April 1999. These were used to estimate indicators for six main groups of variables: population estimates, mortality rates, Census data, benefits data, practice characteristics, and hospital admissions. The data were used to generate a number of population and practice variables describing the characteristics of each primary care group in London. We also obtained information at ward level by age and sex on the number of patients admitted for coronary artery bypass grafting (CABG) and angioplasty (PTCA); cataract surgery; and hip replacement in patients aged 65 years and over during the period 1997-1998. These data were used with population estimates to estimate directly age-standardized annual operation rates for each primary care group for the study period. RESULTS: Operation rates for the three groups of procedures examined in this study varied widely between the 66 primary care groups. The directly standardized combined CABG and PtCA rate varied from 28 to 193 per 100,000 (mean 80); cataract surgery rates from 167 to 618 per 100,000 (mean 291); and hip replacement in people aged 65 years and over from 74 to 363 per 100,000 (mean 202). Correlations between operation rates and population and practice variables were relatively weak. Multifactorial models explained 57 per cent of the variation in CABG and PTCA rates, 34 per cent of the variation in cataract surgery rates, and 27 per cent of the variation in hip replacement rates. CONCLUSIONS: There are wide variations in the operation rates for important groups of surgical procedures among primary care groups. These variations are poorly explained by differences in the population and practice characteristics of primary care groups. The underlying reasons for variations in operation rates need to be better understood if the ambitious targets for access to elective surgery outlined in the NHS plan are to be met. PMID- 11939380 TI - An epidemic pattern of murder. AB - The dates and places of 105 suspected or proven murders committed by Dr H. Shipman of Hyde in Greater Manchester between 1984 and 1998 were subjected to epidemiological analysis. These interim data were assembled through a variety of media sources and were based upon court proceedings and police investigations. The analysis revealed a significant excess of case pairs separated jointly by distances <0.6 km and by times <7 days. These sequences were also associated with purely geographical groupings within spatial diameters <0.2 km, but with longer time intervals. This was confirmed by an extended analysis of case triplets occurring within short times and distances. They showed a significant excess. These clusters were probably generated by a combination of psychopathic obsession, of local opportunity, and of caution; and in similar circumstances could occur again. The findings demand the regular display of mortality data in a format permitting the intuitive recognition of similar phenomena elsewhere. PMID- 11939381 TI - Randomized controlled trial to assess the effectiveness of a primary health care liaison worker in promoting shared care for opiate users. AB - BACKGROUND: Recent national guidelines emphasize the requirement for all general practitioners to manage drug users within a shared care scheme and suggest that a primary health care liaison worker (PHCLW) may facilitate these arrangements. We undertook a group-randomized, randomized controlled trial to determine the effectiveness of a PHCLW in promoting shared care. METHODS: Primary health care teams in Stockport Health Authority, North West England, were randomly allocated to either an intervention arm, who were offered the services of a PHCLW, or to a control arm, who were offered standard support from the community drug team (CDT). The proportion of CDT clients with a history of regular opiate misuse who were in shared care 12 months after randomization was compared across study arms. RESULTS: Eighteen (24.0 per cent) of the 75 CDT clients in the intervention arm but none of the 80 CDT clients in the control arm were in shared care at 12 months (chi2 = 9.37, df = 1, p < 0.01; 95 per cent confidence interval 8.6-39.4 per cent). CONCLUSION: A PHCLW can significantly increase the number of CDT clients in shared care arrangements. PMID- 11939382 TI - Primary care teams work harder in deprived areas. AB - BACKGROUND: The NHS Plan promises an equitable distribution of resources within primary care. To inform the debate on the extent to which resources should be redistributed we examined the association between primary care activity and deprivation. We used the natural experiment of the organization of primary care in Mansfield, Nottinghamshire, where town centre general practices have patients from electoral wards with a range of socio-economic characteristics who are subject to the same degree of supplier-induced demand and variations in data quality. METHODS: We used one year's prospective data for two practices with 20,106 patients from 15 electoral wards. We performed linear regression analysis of directly age-standardized rates for different types of primary care activity and primary care morbidity-specific contacts against Townsend and Index of Multiple Deprivation 2000 scores. RESULTS: There were 44 per cent more out-of hours contacts in more deprived areas (95 per cent confidence interval (CI) 17-70 per cent), 18 per cent more surgery consultations (95 per cent CI 8-27 per cent), and 28 per cent more same-day consultations (95 per cent CI 12-44 per cent). Routine visits by doctors and contacts by district and practice nurses did not have substantial associations with deprivation. Morbidity-specific contacts for psychological problems and respiratory problems were associated with deprivation but there was no significant association for contacts for low back pain, asthma or menopausal problems. CONCLUSIONS: Different types of primary care activity and contacts for different morbidities had different associations with deprivation. This makes it difficult to recommend a simple list size adjustment; however, increased activity in deprived wards needs to be recognized in resource allocation, service configuration and performance management in primary care. PMID- 11939384 TI - Special status consultation for frequent attenders. Who are the candidates? AB - BACKGROUND: The need to optimize frequent attender (FA) care and to economize on scarce resources in primary care has given rise to calls for intervention. However, precise knowledge of the specifics of this need is scarce, as is knowledge of how best to orchestrate intervention vis-a-vis particular FA groups. The aim of the study was to analyse the need for a special FA status consultation and to analyse if such need springs from physical, mental or social factors among FAs. METHOD: Half of the 270 general practices in the County of Aarhus, Denmark, were randomly allocated to the study group (in total 132 practices; 220 general practitioners (GPs)). From these practices age- and gender-stratified samples of FAs were drawn. FAs were defined as the top 10 per cent attenders in 12 months. Questionnaires were used to elicit FA and GP attitudes to the introduction of a status consultation and to obtain GPs' evaluation of their FAs' physical, mental and social status. The special FA consultation was introduced as a reimbursed half-hour consultation where the FA and the GP could discuss the patient's current and future care status. RESULTS: Over three-quarters of the FAs would accept the offer to have the status consultation, and the GPs would recommend it for one-fourth of their FAs. The expressed need was highly associated with an expected positive effect on health (prevalence ratio (PR) for GPs: 11.7 (8.9 15.5), PR for FAs: 1.6 (1.5-1.8)) and a decrease in attendance (PR for FAs: 1.4 (1.3-1.5)). The GPs quoted physical, mental and social indications for the consultation. The FAs who had high levels of functional symptoms were most likely to accept the consultation. CONCLUSIONS: There was an expressed need for an FA status consultation, which may lead to a positive effect on health and attendance. The GPs pointed to physical, mental and social indicators for a special FA consultation. PMID- 11939383 TI - Management of tuberculosis in a British inner-city population. AB - BACKGROUND: The aim of the study was to investigate the management of patients with tuberculosis (TB) in terms of their utilization of health service resources. METHODS: An analysis of patient records was carried out in an NHS Trust in East London, United Kingdom, serving a socioeconomically deprived population. The subjects were all residents of Tower Hamlets treated for drug-sensitive TB in the in-patient and out-patient departments of the Trust in 1998. RESULTS: Of the 62 patients with TB studied, 38 (61 per cent) had an in-patient stay at some stage of their management. Twenty-six of these 38 were admitted acutely ill via the Accident and Emergency Department, 16 having self-presented and 10 after urgent referral by their general practitioner. Only four of the total 62 patients were admitted with previously diagnosed disease, and all four had significant complications necessitating admission. Eight patients were admitted electively for investigation, typically being brief admissions for surgical biopsy. Median in-patient stay was 14 days (range 1-144 days), and in six cases we identified potentially remediable delays in diagnosis and initiation of therapy. CONCLUSIONS: UK and US guidelines for TB imply out-patient management as the norm. Our study shows a very high rate of in-patient care, largely a consequence of the emergency admission of acutely ill, previously undiagnosed cases. There are public health implications in terms of spread of infection from individuals with advanced disease. The high utilization of expensive in-patient resources has significant implications for purchasers and providers of care for TB in socio economically deprived areas. Further, the cost-effectiveness of public health interventions aimed at limiting the spread of TB should be assessed by reference to this true, high cost of managing TB, not a low cost based on false assumptions about rates of out-patient versus in-patient care. PMID- 11939385 TI - An analysis of predictors of success in Part II MFPHM. Membership of the Faculty of Public Health Medicine. AB - BACKGROUND: The examination for Part II of the Membership of the Faculty of Public Health Medicine (MFPHM) has been held since 1974. Candidates and examiners have hypothesized a number of factors as likely to increase the chances of success in the exam. This study formally investigates factors which may predict a candidate's success at their first attempt. METHODS: Routinely available data on candidates sitting the exam from March 1996 to June 1999 were examined initially by univariate analysis. A logistic regression model was then constructed entering all variables identified as statistically significant as well as the following variables initially thought to be relevant (a priori hypotheses): age, sex, university versus NHS training location; splitting competencies evenly across two reports; and training within North Thames deanery. RESULTS: Four factors were identified as statistically significant within the logistic regression model. These were: possession of either MRCP (odds ratio (OR) of failure = 0.33, p = 0.003) or MRCGP (OR = 0.45, p = 0.016), success at UK Part I at first attempt (OR = 0.42, p = 0.014), and North Thames training (OR = 0.44, p = 0.048). CONCLUSION: Possession of MRCP or MRCGP, or passing UK Part I MFPHM are factors not normally open to change by candidates once they begin working towards Part II MFPHM. Training in North Thames was also identified as an independent predictor of success. It would seem advisable to investigate in further detail whether candidate selection (not adjusted for in the model) or aspects of the North Thames training programme are responsible for this observed phenomenon. PMID- 11939386 TI - Public health medicine: the constant dilemma. AB - There is a well-known quotation by the nineteenth-century sociologist Virchow (quoted in Ref. 1) that aptly captures the dilemma that has confronted public health medicine since the specialty was created as a discrete entity in 1848. Virchow said: 'Medicine is politics and social medicine is politics writ large!' What does this mean in relation to effective public health medicine practice and how is it likely to affect its future? There is increasingly limited freedom of expression within the current context of political correctness, central control and a rapidly burgeoning litigious climate. The purpose of this paper is to explore these issues and to propose a means of maintaining public health medicine integrity within a working environment where action is becoming rapidly constrained by political rigidity. An additional factor to be included in the dialogue is the current context within which public health physicians work. Because the majority of public health doctors are employed within the National Health Service (NHS), they are finding themselves being expected to take on tasks and responsibilities marginal to their essential purpose and function. For example, public health physicians spend a great deal of time involved in detailed deliberations about health service provision. Although there is a great deal of evidence to show that good quality health care provision positively affects the health of the individual, there is no evidence to show that this activity has any effect on the population's health status. The essence of public health medicine practice is the prevention of ill-health and the promotion of the health of the population and, consequently, attention needs to be focused on the root causes of disease. However, as these are outside the aegis of the NHS, public health medicine involvement in such issues as education, nutrition, housing, transport and poverty is regarded as marginal to the NHS corporate agenda. PMID- 11939387 TI - Quarterly communicable disease review. July to September 2001. PMID- 11939388 TI - NHS direct--a new paradigm of medical care. PMID- 11939389 TI - Burden of respiratory syncytial viral infections on paediatric hospitals: a two year prospective epidemiological study. AB - This study was designed to prospectively investigate the hospitalisation patterns for respiratory syncytial virus (RSV) disease in children up to 2 years of age, both healthy full-term children and high-risk children. The aim was to evaluate children hospitalised for a respiratory tract infection in four regions and seven paediatric hospital departments or clinics in Greece and to estimate the burden of RSV disease on paediatric hospital beds, as well as to determine the target high-risk age group over two consecutive respiratory infection seasons. Among a total of 1,710 children, the overall prevalence of RSV disease was 33.1%, but prevalence differed between seasons (27% vs. 37.7%, P<0.01). The peak prevalence of RSV disease occurred in February, with the onset in November and the end in May. The monthly distribution was similar in all regions. RSV positivity was strongly associated with young age (4.7 vs. 6.6 months, P<0.001); classification within a high-risk group (43% vs. 32.6%, P=0.04); lower respiratory tract infection (93.8% vs. 86%, P=0.001), particularly bronchiolitis (80.9% vs. 63.9%, P=0.001); more severe disease (P<0.001); longer stay in hospital (P<0.001); and poorer outcome (P<0.001). RSV infections accounted for 38% of the total hospitalisation days for all respiratory infections, and 77% of the hospitalisation days for RSV infection were for infants less than 6 months of age. PMID- 11939390 TI - Investigation of an outbreak due to Alcaligenes xylosoxydans subspecies xylosoxydans by random amplified polymorphic DNA analysis. AB - In 1999, over a 3-week period, Alcaligenes xylosoxydans subsp. xylosoxydans was isolated from five blood cultures and one cerebrospinal fluid specimen from five children hospitalized in a pediatric hematology ward as well as from two respiratory therapy devices of two children hospitalized in an intensive care unit. The infection control unit of the hospital conducted an epidemiological investigation and identified a detergent-disinfectant solution as the source of contamination. Conventional biochemical tests, antimicrobial susceptibility tests and random amplified polymorphic DNA (RAPD) fingerprinting were used to compare clinical and environmental isolates. RAPD analysis proved to be more discriminant than biotyping or antibiotyping in this context and identified the common source of the outbreak. PMID- 11939391 TI - Two-year population-based molecular epidemiological study of tuberculosis transmission in the metropolitan area of Milan, Italy. AB - A 2-year, population-based, molecular epidemiological study was conducted in Milan, Italy, to determine the proportion of tuberculosis (TB) cases attributable to recent transmission. All strains were typed by restriction fragment length polymorphism (RFLP) analysis; clustering was considered indicative of recent transmission. Of the 581 cases, 239 (41.1%) belonged to clusters that consisted of 2 to 11 patients; 28.1% were attributable to recent transmission (number of clustered patients minus 1). Clustering was associated with multidrug-resistant Mycobacterium tuberculosis strains (74.2% of cases), AIDS (60.2%), and a history of incarceration (67.4%). The frequency of multidrug-resistant Mycobacterium tuberculosis was 5.3% overall (15.4% among AIDS patients). Among AIDS patients, infection with a resistant strain was independently associated with clustering (odds ratio, 1.32; 95% confidence interval, 1.07-1.163), while among non-AIDS patients, three factors were associated with clustering: history of incarceration (odds ratio, 2.03; 95% confidence interval, 1.41-2.92), age <30 years (odds ratio, 1.43; 95% confidence interval, 1.05-1.94), and native-born Italian nationality (odds ratio, 1.44; 95% confidence interval, 1.08-1.92). Of the 118 patients who belonged to either the smallest or the largest cluster, 19 (16.1%) reported an epidemiological link with another study patient. The results of this study highlight the need for control programs that focus on selected high-risk groups consisting primarily of HIV-infected individuals and persons with social and lifestyle risks for TB. These programs should be aimed at reducing the probability of transmission of drug-resistant TB through early identification of cases and provision of effective treatment until the individual is cured. PMID- 11939392 TI - Strain variation in the MRSA population over a 10-year period in one Dublin hospital. AB - Between 1989 and 1998, the number of patients carrying methicillin-resistant Staphylococcus aureus in one Dublin hospital increased fourfold, and the antibiogram-resistogram (AR) type distribution changed. In 1989, the predominant AR types were AR01 and AR02; in 1993, AR14 predominated; and in 1994, AR14 and AR13 were predominant. By 1998, the prevalence of AR13 and AR14 had declined and AR06 and AR07 were observed more frequently. In 1989, 65% of isolates were nontypeable using the International Basic Set of Typing Phages. This percentage increased to 78% in 1998. Total cellular DNA macrorestriction analysis reflected the changing AR type distribution. No vancomycin-intermediate isolates were recovered, but possible heteroresistance was observed in 2.7% of isolates. High level mupirocin resistance occurred in 4% of isolates, and 32% exhibited low level resistance. PMID- 11939393 TI - Emergence of cotrimoxazole- and quinolone-resistant Campylobacter infections in bone marrow transplant recipients. AB - Clinical and microbiological data were collected prospectively from 704 patients who underwent bone marrow transplantation (BMT) during an 11-year period (1991 2001), and the first two cases of Campylobacter infection occurring in BMT recipients in the pre-engraftment period were identified. The two cases occurred on days 2 and 3 post-BMT, respectively. Both patients had Campylobacter jejuni enteritis, and one case was complicated by bacteraemia. In both cases the presenting symptoms were indistinguishable from hospital-acquired pre-engraftment diarrhoea, which is commonly caused by Clostridium difficile. Both of the Campylobacter jejuni isolates were resistant to cotrimoxazole and ciprofloxacin. Both patients responded to intravenous meropenem and subsequently had uneventful marrow engraftment. PMID- 11939394 TI - False-positive results caused by cotton swabs in commercial Aspergillus antigen latex agglutination test. AB - While testing for Aspergillus spp. using a commercially available latex agglutination test on a brain biopsy wrapped in a cotton swab, false-positive results were obtained. Subsequent application of the test on lavage samples obtained using cotton and synthetic swabs resulted positive with the cotton swabs only. This suggests that epitopes cross-reactive with Aspergillus galactomannan may be present in cotton. PMID- 11939395 TI - Outbreak of African tick-bite fever in six Italian tourists returning from South Africa. AB - In May 1999, a cluster of cases of African tick-bite fever was detected in six Italian tourists who had returned from South Africa. All of the patients had moderate fever and cutaneous eschars. Regional lymphangitis was observed in three of the patients and skin rash in two. By comparing the number of eschars with the number of detectable bite sites it was suggested that at least two-thirds of the biting vectors were capable of transmitting Rickettsia africae. The clinical course of disease was mild in all cases, and all but one of the patients recovered spontaneously before antibiotic treatment was initiated. The diagnosis of African tick-bite fever was confirmed serologically using both microimmunofluorescence and Western blot tests. PMID- 11939396 TI - Impact of highly active antiretroviral therapy on fever of unknown origin in HIV infected patients. AB - In order to assess the impact of highly active antiretroviral therapy (HAART) on the frequency and etiology of fever of unknown origin in HIV-infected patients, a retrospective study was performed on cases of fever of unknown origin observed from January 1997 to December 1999 in seven hospitals in five cities in Andalusia, Spain. During the period specified, a total of 4,858 HIV-infected patients receiving HAART and 2,787 HIV-infected individuals not receiving HAART were studied. The frequency of fever of unknown origin was 0.6% in patients receiving HAART and 3% in non-HAART patients (P=0.001). Human immunodeficiency virus infection was the only cause of fever found more frequently in the non HAART patient group (P=0.07). The study findings suggest that the use of HAART has reduced the frequency of fever of unknown origin in HIV-infected patients, but the etiology of the condition remains mostly unchanged. PMID- 11939397 TI - Streptococcus agalactiae (Lancefield group B) bacteraemia in nonpregnant adults. PMID- 11939398 TI - Detection of rifabutin resistance and association of rpoB mutations with resistance to four rifamycin derivatives in Helicobacter pylori. PMID- 11939399 TI - Relationship between differences in 16-, 14-, and 15-membered ring macrolides and Streptococcus pyogenes resistance. PMID- 11939400 TI - Cross reaction between a pan-Candida genus probe and Fusarium spp. in a fatal case of Fusarium oxysporum pneumonia. PMID- 11939401 TI - Drug toxicity or syndrome of immune restoration causing fulminant cirrhosis after HAART-induced immune recovery. PMID- 11939402 TI - A case of multiple brain abscesses due to Streptococcus constellatus. PMID- 11939403 TI - Molecular virulence attributes and occurrence of pYV-bearing strains among human clinical isolates of Yersinia enterocolitica in Poland. PMID- 11939404 TI - Worldwide variation in the incidence of Haemophilus influenzae type b meningitis and its association with ampicillin resistance. AB - Some areas of the world are known to have a low incidence of Haemophilus influenzae type b meningitis, although the reasons for this are unknown. Furthermore, no complete evaluation of the worldwide variation in the incidence of Haemophilus influenzae type b meningitis has been published. In the current study, the published medical literature was reviewed to identify all studies conducted in the absence of routine childhood Haemophilus influenzae type b conjugate vaccination that reported an incidence of Haemophilus influenzae type b meningitis among children less than 5 years of age. To test the hypothesis that antibiotic use may have influenced the incidence of meningitis, incidence rates were correlated with antibiotic resistance. Seventy-one articles reported an incidence of childhood Haemophilus influenzae type b meningitis (median, 21 cases per 100,000 population per year; range, 1-95 cases per 100,000 population per year), with Asia and central/southern Europe reporting lower incidences than other areas (median, 5 and 11 cases per 100,000 per year, respectively). Within these regions of low incidence, the proportion of cerebrospinal fluid specimens that had a leukocyte count or glucose or protein level suggestive of bacterial meningitis but from which no organism was identified was low, indicating that there was no large reservoir of Haemophilus influenzae type b meningitis that went undetected by the laboratory. Study-specific incidence rates of meningitis correlated with the proportion of isolates resistant to ampicillin (or producing beta-lactamase) (R2=0.35. P=0.0014). The incidence of Haemophilus influenzae type b meningitis is substantially lower in some areas of the world than others, but this difference is unlikely to be related primarily to laboratory methodology. In contrast, antibiotic use may contribute to the observed differences in incidence. PMID- 11939405 TI - Risk factors for nasal carriage of Staphylococcus aureus in infectious disease patients, including patients infected with HIV, and molecular typing of colonizing strains. AB - Nasal carriage is an important risk factor for Staphylococcus aureus infection, particularly in HIV-infected individuals. In this analytical cross-sectional study, a variety of probable risk factors associated with nasal carriage of Staphylococcus aureus were investigated. HIV-infected patients were examined within a larger cohort of infectious diseases patients. Staphylococcus aureus strains from HIV-infected and non-HIV-infected carriers were identified by molecular biological analysis. One hundred seventy infectious disease patients, 47 of them infected with HIV, were included. All patients were admitted to the University Hospital of Vienna, Austria, between January and July 1999. Independent significant effects on Staphylococcus aureus nasal carriage were found to be HIV status (OR 2.5, 95% CI 1.1-5.6; P=0.0303), history of operation or severe wound within 3 months prior to admission (OR 4.0, 95% CI 1.3-13.0; P=0.0208), presence of an intravenous device within 2 weeks prior to admission (OR 10.8, 95% CI 2.0-59.4; P=0.0065), and intake of antibiotics within 2 weeks prior to hospitalisation (OR 0.2, 95% CI 0.09-0.6; P=0.0016). Molecular analysis of the Staphylococcus aureus strains revealed that the strains in both groups resembled those of healthy nonhospitalized carriers in the community. PMID- 11939406 TI - Relationship of Ureaplasma urealyticum biovar 2 to nongonococcal urethritis. AB - The aim of this study was to determine the frequency of Ureaplasma urealyticum biovars 1 and 2 among 340 men with or without nongonococcal urethritis (NGU) attending a venereal disease clinic in Sweden. NGU was defined by the presence of at least four polymorphonuclear leukocytes per microscopic field (x1,000 magnification) on a smear in which Neisseria gonorrhoeae could not be detected. Ureaplasma urealyticum was detected by polymerase chain reaction, and biovar determination was performed directly on the amplicons by liquid hybridization with biovar-specific probes. Patients with NGU were younger, had had more sexual partners, and exhibited symptoms of urethritis more often than patients without NGU. Ureaplasma urealyticum was detected with the same frequency among patients with and among those without NGU. Among patients with NGU, Ureaplasma urealyticum positive men were more frequently symptomatic than Ureaplasma urealyticum negative men. Among patients without NGU, Ureaplasma urealyticum-positive men had had more sexual partners than Ureaplasma urealyticum-negative men. Ureaplasma urealyticum biovar 2 was detected more often among patients with NGU than among those without (P=0.012). Logistic regression analysis was performed using detection of biovar 2 as the response variable and the following four variables as explanatory variables: presence or absence of NGU, symptoms of urethritis, number of partners, and age < or = 24 years. The only association found was that between Ureaplasma urealyticum biovar 2 and age < or = 24 years. More studies should be conducted to determine the possible pathogenic impact of Ureaplasma urealyticum biovar 2. PMID- 11939407 TI - Wilms' tumor: a paradigm for the new genetics. AB - Carcinogenesis can be triggered by a diverse range of molecular lesions, a variety of which can be illustrated by Wilm's tumor (WT), a pediatric kidney cancer. Molecular defects observed in WTs include several independent targets and mechanisms best exemplified by changes on the short arm of chromosome 11. This article will review the molecular pathology of WT and emphasize the broader ramifications for cancer genetics. Consideration will be given to carcinogenic pathways, novel cellular molecules, and technologies that will assist in the rapid interpretation and assimilation of DNA sequence data arising from the sequencing of the human genome. PMID- 11939408 TI - Novel mdm2 splice variants identified in pediatric rhabdomyosarcoma tumors and cell lines. AB - Mdm2 is an oncogene that binds to and inactivates the tumor suppressor p53. However, the presence of oncogenic splice variants of mdm2 in human tumors that lack the p53 binding site has suggested a p53-independent transforming function for this protein. This report describes expression of 11 different mdm2 splice variants in pediatric rhabdomyosarcoma (RMS) cell lines and tumors at a frequency of 75% and 82%, respectively. Five of these isoforms have previously been described in other tumor histiotypes but six are novel and may be unique to RMS. There was no association between expression of splice variants and mdm2 gene amplification or p53 status. In addition, the frequency of splice variants was much higher than the incidence of mdm2 amplification or p53 mutations. These variants may be important to consider with respect to RMS tumor progression and therapeutic response. PMID- 11939409 TI - Mutation and functional analysis of IL-13 receptors in human malignant glioma cells. AB - We have previously demonstrated that human brain tumor cells, in particular glioblastoma multiforme (GBM), express abundant receptors for interleukin-13 on the cell surface. These receptors are composed of IL-13 receptor (IL-13R)alpha1, IL-13Ralpha2, and IL-4Ralpha chains. The significance of overexpression of IL-13R on tumor cells is not known. Because expression of IL-13R on glioma cells is an unexpected phenomenon, we examined whether these receptors are polymorphic. Therefore, we analyzed cDNA for IL-13Ralpha1 and IL-13Ralpha2 chain genes by PCR based single-strand conformation polymorphism and direct sequencing techniques for a possible polymorphism in 19 GBM, one normal human astrocyte, and two fibroblast cell lines. All analyzed samples except normal astrocytes overexpressed IL-13Ralpha2; however, none of these cell lines showed a mutation in cDNA for IL-13Ralpha2 chain. In contrast, all GBM samples, normal astrocytes, and fibroblasts expressed mRNA for IL-13Ralpha1 with apparent single nucleotide polymorphism in the transmembrane domain. To study the function of IL-13R on brain tumor cells, we investigated the regulation of adhesion molecules by IL-13 as assessed by flow cytometric analysis. A172 cell line expressed a low level of vascular cell adhesion molecule-1 (VCAM-1), while U251 and LA1-5g cell lines expressed intercellular adhesion molecule-1 (ICAM-1). On the other hand E selectin was not expressed in any cell lines. Interestingly, IL-13 increased the expression level of VCAM-1 in A172 cell line in a dose- and time-dependent manner. However, IL-13 did not modulate any other adhesion molecules. These results suggest that IL-13R on GBM cells are not rearranged but appear to be functional. PMID- 11939410 TI - Involvement of claudin-1 in the beta-catenin/Tcf signaling pathway and its frequent upregulation in human colorectal cancers. AB - Accumulation of beta-catenin in cytoplasm and nuclei is frequently observed in a wide variety of tumors arising, for example, in the colon, liver, uterus, or brain. In association with Tcf/LEF transcription factors, beta-catenin regulates expression of genes involved in the Wnt/wingless signaling pathway, but the precise mechanisms are unclear. Here we report evidence that the claudin-1 (CLDNI) gene is one of the genes regulated by beta-catenin. Not only did expression of CLDN1 decrease significantly in response to reduction of intracellular beta-catenin by adenovirus-mediated transfer of wild-type APC into the APC-deficient colon cancer cells, but also two putative Tcf4 binding elements in the 5' flanking region of CLDN1 were confirmed to be responsible for activating its transcription. We documented increased expression of CLDN1 in all 16 primary colorectal cancers we examined, compared with adjacent noncancerous mucosae. Furthermore, immunohistochemical staining demonstrated that claudin-1 was weakly stained at apical boarder of lateral membrane of noncancerous epithelial cells and that it was strongly stained at all cell-cell boundaries and in the cytoplasms of cancer cells. Our results imply that claudin-1 is involved in the beta-catenin-Tcf/LEF signaling pathway, and that increased expression of claudin-1 may have some role in colorectal tumorigenesis. PMID- 11939411 TI - Antitumor activity of ER-51785, a new peptidomimetic inhibitor of farnesyl transferase: synergistic effect in combination with paclitaxel. AB - Inhibitors of ras farnesylation have been extensively studied in the preclinical stage, and some of them are being developed in the clinic. Herein, we describe the antitumor activity of a new farnesyl transferase inhibitor, ER-51785. In vitro, ER-51785 selectively inhibited farnesyl transferase activity (IC50 = 77 nM) compared with geranylgeranyl transferase I activity (IC50 = 4200 nM). In cells, ER-51785 inhibited posttranslational processing of H-ras with IC50 = 28 nM, but not that of rap 1A at concentrations up to 50 microM. This compound also strongly inhibited colony formation of H-ras-transformed NIH 3T3 fibroblasts and EJ-1 bladder carcinoma cells. In vivo, ER-51785 showed potent tumor regression activity against EJ-1 xenografts but only modest activity against MIA PaCa-2 xenografts. Treatment of ER-51785 in combination with paclitaxel exhibited synergistic effects against colony formation and tumor growth of MIA PaCa-2 cells. The results presented herein support the idea that farnesyl transferase inhibitors alone and in combination with other chemotherapeutic agents have the potential to be developed as therapies for tumors expressing H-ras or K-ras oncogenes. PMID- 11939412 TI - Antisense of ATP synthase subunit e inhibits the growth of human hepatocellular carcinoma cells. AB - Differentially expressed genes between normal and hepatocellular carcinoma tissues were investigated using differential display. We identified a cDNA fragment that was overexpressed in cancer tissue. Homology analysis showed that the sequence was identical to human ATP synthase subunit e (hAS-e). Moreover, Northern blot analysis demonstrated that hAS-e was overexpressed in 10 of 11 (91%) specimens of hepatocellular carcinoma compared with corresponding normal tissues. We introduced antisense hAS-e into a human hepatocellular carcinoma BEL 7404 cell and found that downregulation of the hAS-e led to cell growth inhibition. It was also found that the antisense transfection could decrease the serum-stimulated activation of mitogen-activated protein kinase (MAP kinase). Together, the results suggest that antisense of hAS-e can inhibit cell proliferation through the MAP kinase pathway. Our data indicate that hAS-e may become a new target in gene therapy. PMID- 11939413 TI - Molecular characterization of the DICE1 (DDX26) tumor suppressor gene in lung carcinoma cells. AB - We have determined the genomic structure of the candidate tumor suppressor gene DICE1 (DDX26). The DICE1 gene colocalizes with microsatellite marker D13S284 telomeric to the RB1 gene in chromosomal region 13q14.3. The DICE1 gene encodes 18 exons that are preceded by a GC-rich promoter region. CpG sites flanking a predicted TATA box were found to be hypermethylated in tumor cells that exhibited decreased DICE1 expression. This suggests tumor-specific transcriptional silencing of the DICE1 gene may occur. Aberrantly spliced products were detected in two of three DICE1 expressing cell lines. The predicted DICE1 amino acid sequence is evolutionarily conserved in mouse, fruit fly (D. melanogaster), and nematode (C. elegans). A DEAD box characteristic of ATP-dependent helicases is the predominant motif found in DICE1 and its mouse and fruit fly homologues. Motifs other than the DEAD box are reminiscent of members of the helicase superfamily II but there is considerable variation from the typical DEAD box helicases. Expression of DICE1 green fluorescent fusion protein showed a preferential localization of DICE1 in the nucleus. This suggests that DICE1 is involved in nuclear processes such as DNA repair, transcription, or RNA splicing. PMID- 11939414 TI - Antitumor effect of VNP20009, an attenuated Salmonella, in murine tumor models. AB - VNP20009, a genetically modified strain of Salmonella typhimurium with deletions in the msbB and purI loci, exhibited antitumor activities when given systemically to tumor-bearing mice. VNP20009 inhibited the growth of subcutaneously implanted B16F10 murine melanoma, and the human tumor xenografts Lox, DLD-1, A549, WiDr, HTB177, and MDA-MB-231. A single intravenous injection of VNP20009, at doses ranging from 1 x 10(4) to 3 x 10(6) cfu/mouse, produced tumor growth inhibitions of 57-95%. Tumor volume doubling time, another indicator for tumor growth inhibition, also significantly increased in mice treated with VNP20009. Using mice with immune system deficiencies, we also demonstrated that the antitumor effects of VNP20009 did not depend on the presence of T and B cells. In addition, VNP20009, given intravenously, inhibited the growth of lung metastases in mice. Only live bacteria showed the antitumor effect. PMID- 11939416 TI - Adolescent perceptions of relevant social problems. AB - PROBLEM: Social problems can be a significant stressor for adolescents. Disturbed youth often experience negative outcomes to social problems, solve problems poorly, and display distortions in reasoning about social problems. METHODS: A series of focus groups was conducted with adolescents (N = 36), and the data was analyzed inductively. Activities to increase the rigor of the study data (establishing credibility, trustworthiness, and transparency) were incorporated into the phenomenological research design. FINDINGS: Five social problem themes with critical attributes were perceived to be relevant to teenagers in today's society. CONCLUSIONS: Adolescents' social problems shift in response to a changing society. Findings from this study can be used to help target assessments and treatment plans when working with disturbed youth. PMID- 11939415 TI - Connectedness of pregnant adolescents who smoke. AB - PURPOSE: To examine the relationship of the constructs from the family and school environments and protective individual characteristics to health risk behaviors among pregnant adolescents who smoke. METHODS: Inner-city adolescents (N = 53), aged 14-19 years, who were participating in a smoking cessation intervention program, completed self-report demographic, social, and behavioral questionnaires. FINDINGS: Independent variables included family connectedness, school connectedness, and individual characteristics of religiosity, self-esteem, and grade-point average. Three areas of health risk behavior were dependent variables: delinquent behavior, substance, and precocious sexual behavior. CONCLUSIONS: Positive self-esteem served as a protectivefactorfor alcohol use. In contrast to past studies, school connectedness was a risk, not a protective, factor for substance abuse. Neither school nor family connectedness were protective against delinquent or precocious sexual behavior. PMID- 11939417 TI - The aftermath of September 11, 2001. PMID- 11939418 TI - Is there a culture of youth violence? PMID- 11939419 TI - Mental health parity--just an illusion? PMID- 11939420 TI - The effects of a stress-management program on self-concept, locus of control, and the acquisition of coping skills in school-age children diagnosed with attention deficit hyperactivity disorder. AB - PROBLEM: What is the effectiveness of a stress-management program on self concept, locus of control, and acquisition of appropriate coping strategies in children with attention deficit hyperactivity disorder (ADHD)? METHODS: Forty-two children with ADHD (age range 9-12) were randomly assigned to a control group, a therapist-led group, or a group in which the stress-management techniques were taught by the parents. The subjects were pre- and posttested on self-concept, locus of control, and acquisition of coping skills. FINDINGS: Although there were no significant changes in any of the three groups for acquisition of coping skills, the children in the therapist-led group reported more appropriate coping strategies. CONCLUSIONS: Involving parents in a cognitive-behavioral approach could improve parent-child relationships and further increase the opportunities for successful academic and social functioning. PMID- 11939421 TI - Understanding the trusted doctor and constructing a theory of bioethics. AB - This paper offers a constructivist account of bioethics as an alternative to previous discussions that explained the ethics of medicine by an extrapolation of principles or virtues from ordinary morality. Taking medicine as a higher and special calling, I argue that the practice of medicine would be impossible without the trust of patients. Because trust is a necessary condition for medical practice, the ethics of the profession must provide the principles for guiding physician behavior and the profession toward promoting trust and being trustworthy. In a phrase, that principle is "seek trust and deserve it." I sketch out how the concept of trust provides a different justification for common sense principles of bioethics and explain how the concept of trust provides reasonable guidance for resolving moral conflicts within medicine. The trust-seeking approach provides a new and unexpected ordering of some traditional medical values, it reveals the weightiness of previously undervalued bioethical precepts, and illuminates the centrality of some largely ignored obligations of medicine. It also has the power to guide clinical practice and to inform the profession about standards for medical institutions. PMID- 11939422 TI - A meta-ethical critique of care ethics. AB - A meta-ethical analysis demonstrates that care ethics is a grounded in a distinct mode of moral reasoning. This is comprised primarily of the rejection of principles such as impartiality, and the endorsement of emotional or moral virtues such as compassion, as well as the notion that the preservation of relations may override the interests of the individuals involved in them. The main conclusion of such a meta-ethical analysis is that such meta-ethical foundations of care ethics are not sound. Reasonable alternatives for care ethics may be its formulation as an additional principle within an established principlist framework, or the move to a dialogical ethics, where the good to be acted upon is not decided in advance but rather critically discussed and established within the encounter of the parties involved. PMID- 11939423 TI - Cultural diversity and patients with reduced capacity: the use of ethics consultation to advocate for mentally handicapped persons in living organ donation. AB - Living organ donation will soon become the source of the majority of organs donations for transplant. Should mentally handicapped people be allowed to donate, or should they be considered a vulnerable group in need of protection? I discuss three cases of possible living organ donors who are developmentally disabled, from three different cultures, the United States, Germany, and India. I offer a brief discussion of three issues raised by the cases: (1) cultural diversity and cultural relativism; (2) autonomy, rationality, and self-interest; and (3) the proper use and role for clinical ethics consults. PMID- 11939425 TI - Self-healing forces and concepts of health and disease. A historical discourse. AB - The phenomenon of self-healing forces has again and again challenged doctors in the different historical periods of medical science. They relied on effects of self-healing forces in diagnosis and therapy. They also tried to explain these effects based on the current model of organism. The understanding of this phenomenon has always influenced the understanding of therapy and played a role in defining the concept of health and disease. In the 17th and 18th century the idea of self-healing force was interpreted as a phenomenon related to the organic forces, whereas in the 19th century the explanation was reduced to a materialistic mechanism. Nowadays the knowledge of heath-shock-proteins open the way of a new understanding of the organic defense mechanisms. PMID- 11939424 TI - Shaping individuality: human inheritable germ line gene modification. AB - In this paper I deal with ethical factors surrounding germline gene therapy. Such implications include intergenerational responsibility, human dignity, moral status of embryos and so on. I will explore the relevance of the above mentioned issues to discuss the ethical implication of human germline gene therapy (HGLT). We will see that most of arguments claimed by bioethicists do not provide valid reason to oppose HGLT. I will propose an alternative view, based on personal identity issues, to discuss the ethics of human inheritable gene modification. PMID- 11939426 TI - The ethics of alpha: reflections on statistics, evidence and values in medicine. AB - As health care embraces the tenets of evidence-based medicine it is important to ask questions about how evidence is produced and interpreted. This essay explores normative dimensions of evidence production, particularly around issues of setting the tolerable level of uncertainty of results. Four specific aspects are explored: what health care providers know about statistics, why alpha levels have been set at 0.05, the role of randomization in the generation of sufficient grounds of belief, and the role of observational studies. The essay concludes with recommendations to acknowledge the value permeation of outcome measures and suggests that attention to reasoning and argument analysis can augment traditional evidence-based approaches in providing a robust critical approach to medical knowledge. PMID- 11939428 TI - Vasopressin and shock. PMID- 11939429 TI - Management of difficult spinal anaesthesia in a patient with adult lumbar scoliosis. PMID- 11939430 TI - Emergency percutaneous dilatational cricothyroidotomy after failed intubation. PMID- 11939431 TI - An unusual case of transient paraplegia. PMID- 11939432 TI - Non-bronchoscopic bronchoalveolar lavage in the microbiological diagnosis of pneumonia in mechanically ventilated patients. AB - A prospective study comparing standardized non-bronchoscopic bronchoalveolar lavage (sNB-BAL) and non-specific endotracheal aspirate (NsETA) in the microbiological diagnosis of pneumonia in mechanically ventilated patients is described. One hundred episodes in 82 mechanically ventilated patients with or without radiological and clinical diagnostic criteria of pneumonia were studied. NsETA and sNB-BAL was performed on the day of study. Fifty-one patients had pneumonia (21 ventilator-associated, 12 hospital-acquired, 18 community-acquired) and 49 had no pneumonia as defined by widely accepted clinico-radiological criteria. The sNB-BAL was found to be significantly more specific (0. 73) compared to NsETA (0.35) for the microbiological diagnosis of pneumonia. Colonization rates with NsETA were significantly higher compared to sNB-BAL (P value <0.0001). No patient had complications attributable to the sNB-BAL procedure. We conlude that sNB-BAL is a safe, effective, sensitive, specific and inexpensive procedure for the serial evaluation of pneumonia in mechanically ventilated patients. PMID- 11939434 TI - Accuracy of portable infusers under hyperbaric oxygenation conditions. AB - Three types of portable infusers with different infusion mechanisms were evaluated with regard to their accuracy during a hyperbaric oxygenation protocoL The power driving the pump is provided by either a balloon, a spring or a vacuum mechanism. Performance during hyperbaric oxygenation (HBO) varied between the devices, probably due to the difference in driving mechanism. Flow delivery by the vacuum type infuser is substantially affected by HBO. Doubling the ambient pressure approximately doubled flow delivery from the vacuum type device. We suggest that other devices are more suitable for use in this clinical situation. We conclude that it is desirable to check the performance of any infuser intended for use during hyperbaric oxygenation and to be mindful of potential differences among such devices. PMID- 11939433 TI - The peripherally inserted central catheter (PICC): a prospective study of its natural history after cubital fossa insertion. AB - A prospective cohort study was undertaken to describe the natural history of the cubital fossa peripherally inserted central catheter (PICC), determine which factors influenced the hazard of complication and develop a standard methodology for evaluation of a PICC service. A total of 4349 patient days of PICC observation were analysed using survival analysis techniques. The median time to PICC removal for a complication was 60 days. The most common complications were phlebitis, malposition and tip migration. Complications usually occurred during the first week. There was only one episode of line-related sepsis. Size 3 French gauge catheters had a complication rate of 7.3 per 1,000 line days compared to 14.2 for 4 French catheters (hazard rate 1.26 90% CI 1.02 to 1.55). PICCs requiring two or more attempts at insertion were more likely to develop complications than those inserted at the first attempt: 20 per 1,000 line days vs 10.5 but the confidence intervals were wide (hazard rate 1.91, 90% CI 0.90 to 4.05). Operator (amongst the four experienced operators who inserted all PICCs), arm of placement, or medial or lateral placement in the cubitalfossa did not influence PICC survivaL PMID- 11939435 TI - Comparison of isoprenaline with adrenaline as components of epidural test dose solutions for halothane anaesthetized children. AB - In this randomized controlled trial we studied the haemodynamic effects of intravenous injection of lignocaine 1 mg/kg with either no additive, isoprenaline 0. 05 microg or 0.075 microg/kg, or adrenaline 0.5 microg/kg in children anaesthetized with halothane. An increase of > or = 10 bpm in heart rate or > or = 20% rise in blood pressure was taken as a positive test response. Adrenaline was confirmed to be an unreliable marker with a positive test in only 67% of cases. For both doses of isoprenaline, the heart rate changes produced a positive test in 100% of cases. There was no statistically significant difference between the two doses of isoprenaline in regard to mean maximum increase in heart rate, mean time of beginning, ending or duration of positive test response and mean time to reach peak heart rate. The positive test response began between 10 and 50 seconds and lastedfor an average of 69 +/- 54 seconds with the lower dose of isoprenaline 0.05 microg/kg No advantage could be seen with the higher dose of isoprenaline. Change in blood pressure was not a useful criterion to detect intravascular injection. Recommendations for the use of isoprenaline as a marker of intravascular injection during regional blockade must be deferred until the safety of this agent's effect on neurological tissue is confirmed. PMID- 11939436 TI - Intravenous induction agents: benzodiazepines. PMID- 11939437 TI - Bispectral index in assessment of adequacy of general anaesthesia for lower segment caesarean section. AB - Awareness among parturients during general anaesthesia for caesarean section, though now uncommon, remains a concern for obstetric anaesthetists. We examined the adequacy of our general anaesthetic technique for avoiding explicit awareness by determining the depth of anaesthesia using Bispectral Index (BIS) monitoring. Twenty ASA1 parturients having general anaesthesia for lower segment caesarean section were studied. The drugs and doses used for each anaesthetic were similar Intraoperative Bispectral Index, haemodynamic parameters, end-tidal isoflurane concentration and inspired nitrous oxide fraction were measured and the postoperative incidence of explicit awareness was assessed. All anaesthetists were blinded to the Bispectral Index value throughout the operation. The depth of anaesthesia at various stages of the operation was evaluated by recording the Bispectral Index. Patients were interviewed for any intraoperative recall or awareness at the end of operation. A median BIS of 70 or below was recorded on most occasions during surgery. The range was 52 to 70, with values reaching 60 and below at intubation, uterine incision and delivery. Haemodynamic stability was satisfactory and there was no case of uterine atony, fetal compromise or postpartum haemorrhage. No patient experienced intraoperative dreams, recall or awareness. Our current general anaesthetic technique appeared inadequate to reliably produce BIS values of less than 60 that are associated with a low risk of awareness. However, no patients experienced explicit awareness. PMID- 11939438 TI - Internal jugular vein patency following carotid endarterectomy. AB - We performed ultrasound examinations on forty-five patients with a history of carotid endarterectomy to assess the patency and position of the internal jugular vein on the side of the endarterectomy. The study was performed in a tertiary referral centre. In the 55 internal jugular veins examined (ten bilateral procedures), the veins were patent and in a normal relationship to the carotid artery. We concluded that previous carotid surgery should not be a contraindication to cannulation of the internal jugular vein. PMID- 11939439 TI - An evaluation of non-invasive blood pressure (NIBP) monitoring on the wrist: comparison with upper arm NIBP measurement. AB - The arm is the traditional site for application of an oscillometric non-invasive blood pressure (NIBP) cuff This study, which compares upper arm NIBP to wrist NIBP, involved 510 same arm sequential paired blood pressure (BP) measurements in 85 volunteers. Wrist NIBP consistently overestimated mean arterial, systolic and diastolic pressure by approximately 10 mmHg. Ninety per cent of the mean arterial pressure differences at the wrist were within a range of +/- 9 mmHg around a mean difference of 10.6 mmHg. The systolic pressure difference was 11.2 mmHg with 90% of differences between +/- 12 mmHg. The diastolic pressure difference was 10.2 mmHg with 90% of differences between +/- 9 mmHg. With the device used and within the normal blood pressure range, compensation can be performed by subtracting 10 mmHg from the measured values or simply by elevating the wrist about 15 cm and taking the BP at face value. Wrist NIBP may be a viable clinical alternative in situations where difficulty occurs with upper arm NIBP measurement. PMID- 11939440 TI - The modified Cormack-Lehane score for the grading of direct laryngoscopy: evaluation in the Asian population. AB - The use of a modified Cormack-Lehane scoring system (MCLS) of laryngoscopic views, as previously introduced in the Western population, was investigated during direct laryngoscopy in the Asian population. We studied the distribution of the different grades of MCLS, the predictive factors and rate of difficult laryngoscopy, and the association with difficult intubation. Six hundred and five patients requiring tracheal intubation during general anaesthesia were prospectively studied. The optimal views during direct laryngoscopy were scored using the 5-grade MCLS system. The distribution of the laryngoscopy scores was 73.9% Grade 1 (full view of the vocal cords), 21.0% Grade 2A (partial view of the vocal cords), 3.3% Grade 2B (only the arytenoids and epiglottis seen), 1.6% Grade 3 (only epiglottis visible) and 0.2% Grade 4 (neither the epiglottis nor glottis seen). External laryngeal pressure was necessary in 45.3% of cases to optimize laryngoscopic views. Grade 2B was associated with significantly higher incidence of difficult intubation compared with Grade 2A (65% vs 13.4%). The rates of difficult laryngoscopy and intubation were 5.1% and 6.9% respectively. The Mallampati classification and thyromental distance were associated with low predictive value for difficult laryngoscopy. The MCLS better delineates the difficulty experienced during laryngoscopy than the original Cormack-Lehane grading PMID- 11939441 TI - Detection and correction of accidental oesophageal intubation during flexible lightwand-guided intubation via the intubating laryngeal mask. AB - In the following two-part study, we determined the efficacy of observation of the light glow to detect correct placement of the tracheal tube after lightwand guided tracheal intubation via the intubating laryngeal mask (ILM). We also determined the efficacy of a protocol to correct oesophageal intubation in this situation. In study 1, 80 ASA 1-3, anaesthetized, paralysed patients were randomly assigned to have a tracheal tube, preloaded with a flexible lightwand, placed into either the trachea (n=40) or oesophagus (n=40) under laryngoscope guidance. A blinded observer experienced with the lightwand technique determined whether oesophageal or tracheal intubation had occurred by observation of the light glow. In study 2, 1,000 patients, 400 of whom were included in a previous study, underwent flexible lightwand-guided intubation via the ILM. Placement of the tracheal tube in oesophagus or trachea was determined by observation of the glow and verified by capnography. Oesophageal intubation was corrected by adjusting the ILM position using a protocol comprising two adjusting manoeuvres. Observation, verification and correction were by experienced anaesthetists conducting the case. In study 1, oesophageal and tracheal intubation was correctly detected in 38/40 (95%) and 37/40 (92.5%) patients respectively. In study 2, oesophageal and tracheal intubation was correctly detected in 55/55 (100%) and 945/945 (100%) patients respectively. In 40/55 (73%) patients, tracheal intubation was successful at the second attempt and in 13/55 (24%) at the third or fourth attempt. In 2/55 (4%) patients, persistent oesophageal intubation occurred. The sensitivity, specificity and positive predictive value were 95%, 92.5% and 92.7% respectively in study 1, and were all 100% in study 2. PMID- 11939442 TI - Diclofenac or paracetamol for analgesia in paediatric myringotomy outpatients. AB - This prospective, randomized, double-blind study compared the analgesic efficacy of oral diclofenac resinate 0.5 mg.kg(-1) with paracetamol 15 mg/kg(-1) for control of postoperative pain in paediatric patients for outpatient bilateral myringotomy and tube insertion. Paracetamol, the most commonly used oral analgesic for paediatric patients, was compared with a new palatable syrup formulation of diclofenac. Sixty-three ASA 1 orA SA 2 children aged one year and above were randomly assigned to receive diclofenac (Group A) or paracetamol (Group B). The study drug was given 30 to 60 minutes before induction of anaesthesia. Anaesthesia was induced with either inhalational sevoflurane or intravenous thiopentone. All subjects received intravenous fentanyl 1 microg/kg( 1) intraoperatively. Postoperative pain was assessed by a blinded observer using the CHEOPS score on eye-opening, and then at 10, 30 and 60 minutes. Children with a CHEOPS score > 7 received further fentanyl 1 microg x kg(-1). The number of cases requiring this "rescue" analgesia was recorded. Both groups were comparable in demographics, induction technique, duration of anaesthesia and time between premedication and induction of anaesthesia. Overall, CHEOPS scores were low for both groups at all times and did not differ between the groups at any time. Twenty per cent of the diclofenac group and 27% of the paracetamol group required rescue analgesia (not statistically significant). The efficacy of diclofenac 0.5 mg x kg(-1) and paracetamol 15 mg x kg(-1) as oral analgesic premedication for BMT was comparable in children receiving an anaesthetic which included intraoperative administration of fentanyl 1 microg x kg(-1). PMID- 11939443 TI - Incidents relating to arterial cannulation as identified in 7,525 reports submitted to the Australian incident monitoring study (AIMS-ICU). AB - Arterial cannulation is frequently used in the critical care environment. Literaturefocuses on insertion techniques and complications. This report utilized data from the Australian Incident Monitoring Study (AIMS-ICU) national database to identify common problems and contributing factors associated with the use and maintenance of arterial lines. A review of narratives, keywords and contributing factors yielded 251 reports outlining 376 incidents. Of these, 15% describing line insertion problems, 66% line use and maintenance problems and 19% patient injuries. Inadequate line securing, accidental line dislodgement, incorrect set up, distal ischaemia and infection featured prominently. As a result of the incident, 49% of patients involved suffered no ill effect, 28% minor physiological complications and 15% suffered major adverse effects. Multiple contributing factors were selected for each report, with lack of knowledge, rule based errors, high unit activity, and lack of support staff or supervision selected most frequently. This study highlights the need to employ meticulous insertion technique, line set-up, securing, frequent line assessment and the early removal of lines no longer essential to patient care. Support and education of staff as well as the development of protocols are important for the safe use of arterial lines. PMID- 11939444 TI - Anaesthesia for endoluminal repair of abdominal aortic aneurysms. AB - An audit of 100 patients undergoing elective abdominal aortic surgery either by open aortic repair (OAR group 50 patients) or endovascular aortic repair (EAR group 50 patients) was undertaken to document changes in anaesthetic technique and perioperative outcome. The data for the OAR group was collected retrospectively and thatfor the EAR group prospectively. Combined general anaesthesia and thoracic epidural anaesthesia was used in 44 of the OAR group whereas lumbar central neural blockade alone was used in 47 of the EAR group. The major differences between the two groups were that intraoperative blood loss was significantly less in the EAR group (OAR 1,674 +/- 1,008 ml, EAR 459 +/- 350 ml, P<0.001) and that no patient in the EAR group required admission to the Intensive Care Unit (ICU), whereas ICU time for the OAR patients was 29 +/- 22 hours. Hospital stay was also significantly different between the two groups (OAR 13 +/- 6 days, EAR 5 +/- 3 days, P<0.001). Major complications occurred in 20patients in the OAR group but only 4patients in the EAR group (P<0.001). EAR reduces blood loss, the requirement for ICU admission, and hospital stay. Central neural blockade is a satisfactory anaesthetic technique for EAR. PMID- 11939445 TI - Systemic reperfusion injury during arm replantation requiring intraoperative amputation. AB - This case describes the attempted replantation of an arm following its traumatic amputation. After a five-and-a-half hour ischaemic time, perfusion of the arm was re-established. However over the ensuing hours, the patient developed hyperkalaemia, acidaemia and severe hypotension requiting high dose inotropic support. Ultimately re-amputation was necessary to treat the systemic effects of the ischaemia reperfusion syndrome. PMID- 11939446 TI - Phaeochromocytoma and acute myocardial infarction. AB - Phaeochromocytoma is uncommonly associated with myocardial infarction. We present a patient who, despite established alpha adrenoceptor blockade, sustained an acute myocardial infarction and was found to have coronary artery disease. Indications for coronary revascularization were not met, and adrenalectomy was successfully performed four weeks later. Factors contributing to the myocardial infarction, the role of beta adrenoceptor blockade, the timing of adrenalectomy and the place of coronary revascularization are discussed. PMID- 11939447 TI - Vibrio vulnificus septicaemia. AB - Vibrio vulnificus is an opportunistic pathogen capable of causing a fulminant septicaemia in susceptible patients. Underlying chronic diseases such as liver impairment and immunosuppression are important factors contributing to the severity of the infection and outcome. Early suspicion and diagnosis with appropriate antibiotic therapy is important as delay can adversely affect outcome. For those who develop tissue necrotizing fasciitis, early surgical debridement is recommended to allow better penetration of antibiotics and also to reduce the severity of the septicaemia. Mortality is quoted as between 50% and 90%. Current antibiotic recommendations are intravenous ceftazidime 2 g tds and doxycycline 100 mg od. PMID- 11939448 TI - Multi-organ failure secondary to catastrophic anti-phospholipid syndrome. AB - We describe the case of a young woman with a known history of thrombocytopenia, who developed respiratory and circulatory failure thought to be caused by sepsis. She subsequently was shown to have high titres of anticardiolipin antibodies as well as lupus anticoagulant. Serological tests for various connective tissue diseases were negative. Multiple cerebral ischaemic lesions led to a poor outcome. This case highlights the potential difficulties in differentiating catastrophic anti-phospholipid syndrome from disseminated intravascular coagulation. PMID- 11939449 TI - Cardiovascular collapse caused by carbon dioxide insufflation during one-lung anaesthesia for thoracoscopic dorsal sympathectomy. AB - Carbon dioxide insufflation into the pleural space during one-lung anaesthesia for thoracoscopic surgery is used in some centres to improve surgical access, even though this practice has been associated with well-described cardiovascular compromise. The present report is of a 35-year-old woman undergoing thoracoscopic left dorsal sympathectomy for hyperhidrosis. During one-lung anaesthesia the insufflation of carbon dioxide into the non-ventilated hemithorax for approximately 60 seconds, using a pressure-limited gas inflow, was accompanied by profound bradycardia and hypotension that resolved promptly with the release of the gas. Possible mechanisms for the cardiovascular collapse are discussed, and the role of carbon dioxide insufflation as a means of expediting lung collapse for procedures performed using single-lung ventilation is questioned. PMID- 11939450 TI - Acute lithium intoxication. AB - We present a case of acute lithium intoxication in a 51-year-old woman on chronic lithium therapy. Her serum lithium level was 10.6 mmol/l 13 hours after ingestion and 5.8 mmol/l at 24 hours. Dialysis therapy was not employed and she recovered well after fluid resuscitation. Serum lithium levels in chronic intoxication are more indicative of intracellular lithium concentration and therefore of clinical toxicity, as opposed to serum lithium levels in acute intoxication. Clinical features of toxicity are more important than a spot lithium level. A combination of clinical toxicity, the duration of exposure and a serial profile of serum lithium levels should guide dialytic therapy for removal of lithium. PMID- 11939451 TI - Awake fibreoptic intubation under remifentanil and propofol target-controlled infusion. AB - We present the first report of the use of remifentanil and propofol target controlled infusion to sedate a patient with a difficult airway undergoing awake fibreoptic intubation. This regimen was rapidly titratable, aided suppression of airway reflexes, maintained patient comfort and cooperation and did not compromise spontaneous respiration. The literature regarding infusion rates and potential complications of this technique is reviewed. PMID- 11939452 TI - Combined spinal epidural anaesthesia for vesico-vaginal fistula repair in an achondroplastic dwarf. AB - A 33-year-old achondroplastic female was scheduled to undergo vesico-vaginal fistula repair by the abdominoperineal route. Preoperative examination suggested a difficult airway so a combined spinal epidural technique was used. Subarachnoid block (sensory loss to T6) was established using 0.5% hyperbaric bupivacaine 1 ml. Anaesthesia was prolonged with incremental doses of epidural bupivacaine 0.5% (total 10 ml) and postoperative analgesia was provided with epidural morphine boluses. PMID- 11939453 TI - Fatal epidural infusion-call for a system-wide change. PMID- 11939454 TI - Release of propranolol and diclofenac from low Mw DL-poly(lactic acid). AB - The controlled release of two drugs, i.e. the sodium salt of diclofenac and propranolol was studied, by using low molecular weight D,L-Poly(lactic acid) as a matrix. Tablets of the above polymer containing those drugs were immersed into buffers with various pH values and delivery was recorded as a function of time, via UV-spectroscopy. The results showed that the polymer is appropriate for such biomedical applications, as generally, it ensures complete drug delivery within 45-60 days, which is acceptable for most cases. On the other hand, the release rate depends on many parameters including the interactions among drug, matrix and the surrounding liquid, which adds complexity to the process and requires careful investigation for proper design of a controlled release system. PMID- 11939455 TI - In vitro interactions of biomedical polyurethanes with macrophages and bacterial cells. AB - Three commercial medical-grade polyurethanes (PUs), a poly-ether-urethane (Pellethane), and two poly-carbonate-urethanes, the one aromatic (Bionate) and the other aliphatic (Chronoflex), were tested for macrophages and bacterial cells adhesion, in the presence or absence of adhesive plasma proteins. All the experiments were carried out on PUs films obtained by solvent casting. The wettability of these films was analysed by measuring static contact angles against water. The ability of the selected PUs to adsorb human fibronectin (Fn) and fibrinogen (Fbg) was checked by ELISA with biotin-labelled proteins. All PUs were able to adsorb Fn and Fbg (Fn > Fbg). Fn adsorption was in the order: Pellethane > Chronoflex > Bionate, the highest Fbg adsorption being detected onto Bionate (Bionate > Chronoflex > Pellethane). The human macrophagic line J111, and the two main bacterial strains responsible for infection in humans (Staphylococcus aureus Newman and Staphylococcus epidermidis 14852) were incubated in turn with the three PUs, uncoated or coated with plasma proteins. No macrophage or bacterial adhesion was observed onto uncoated PUs. PUs coated with plasma, Fn or Fbg promoted bacterial adhesion (S. aureus > S. epidermidis), whereas macrophage adhered more onto PUs coated with Fn or plasma. The coating with Fbg did not promote cell adhesion. Pellethane showed the highest macrophage activation (i.e. spreading), followed, in the order, by Bionate and Chronoflex. PMID- 11939456 TI - Performance modification of chitosan membranes induced by gamma irradiation. AB - Trauma of the nervous system often results in permanent functional loss because the spontaneous regeneration of nerves is very difficult. Thus, various methods have been developed to facilitate the restoration of damaged nerve. The biodegradable nerve conduit is one of the most promising methods for nerve regeneration. Chitosan, a natural polysaccharide that has excellent biocompatibility and biodegradability, can be used as conduit material. But, nerves regenerated by nerve conduits made from chitosan have some problems, for example, with their mechanical properties. This article shows that the mechanical properties of chitosan film were markedly improved by selected doses of gamma radiation and cell culturing experiments on the surface of the irradiated chitosan film indicated that the film still has excellent biocompatibility. PMID- 11939457 TI - Gelatin microspheres and sponges for delivery of macromolecules. AB - Gelatin microspheres and gelatin sponges were prepared by coacervation and freeze drying techniques, respectively. Both systems were crosslinked with glutaraldehyde. The mean diameter of the microspheres were in the range of 40-80 microm and the mean pore size of the sponges was 130-220 microm depending on the preparation conditions. Bovine serum albumin (BSA) was added into the preparation solutions and entrapped in the microspheres and sponges. BSA addition to sponges was also achieved by addition of BSA-containing microspheres into the sponges. The release kinetics of BSA from the prepared systems were examined. Studies demonstrated that release is dependent on the amount of BSA present in the system and crosslinking densities of microspheres. It was concluded that gelatin microspheres and gelatin sponges are promising carrier matrices for macromolecules. PMID- 11939458 TI - Quality of life for individuals with SCI: let's keep up the good work. PMID- 11939459 TI - New avenues: exploring approaches for gene therapy in the injured spinal cord. PMID- 11939460 TI - Objective and subjective handicap following spinal cord injury: interrelationships and predictors. AB - OBJECTIVE: To investigate the relationship between objective and subjective indicators of handicap or community participation among people with spinal cord injury (SCI) 1 year postinjury. DESIGN: Longitudinal correlational study of quality of life indicators linked to Northern New Jersey SCI System database. SETTING AND PARTICIPANTS: A total of 126 (80% male) participants was assessed at 1 year post-SCI. Age ranged from 14-83 years (median age = 34 years); 47% had tetraplegia, and 53% had paraplegia. PRIMARY MEASURES: Objective (or normative) handicap was measured using the Craig Handicap Assessment and Reporting Technique and the Community Integration Questionnaire. Subjective feelings about each area of handicap or community participation were assessed using the Andrews Delighted Terrible scale. RESULTS: Significant correlations were found between objective and subjective indicators of handicap in work life, social life, mobility, and economic status. Subjective handicap also correlated modestly with severity of impairment and length of hospitalization. These correlations were, however, weak or inconsistent across individuals. Subjective quality of life was not related to preinjury economic or social handicap. Some participants spontaneously reported dissatisfaction with items outside of the standard outcomes scales used (eg, sexuality and personal relationships). CONCLUSIONS: The weakness and inconsistency of relationships between objective and subjective appraisals of areas of community participation is a challenge to outcomes measurement and has implications for the targeting of interventions. More research is needed to understand relationships between objective indicators of community participation and subjective appraisals of these areas. PMID- 11939461 TI - Gene therapy for spinal cord injury and disease. AB - An incomplete understanding of the pathological processes involved in neurodegeneration and dysfunction of spinal cord injuries and diseases makes these disorders difficult to treat. Repair of damaged or genetically impaired spinal cord also has been limited by the complexity, cellular heterogeneity, and relative inaccessibility of the tissue. Thus, therapeutic options for the treatment of either chronic spinal cord diseases such as amyotrophic lateral sclerosis or acute spinal cord injuries have been rather limited. Potential new therapeutic targets are being identified as our understanding of the molecular pathology involved in neural injury and regeneration increases. Recent advances in gene transfer techniques have made gene therapy a more realistic and viable strategy for the treatment of a broad range of spinal cord disorders. This review summarizes the current state of knowledge regarding the limitations and recent advances in gene therapy and potential application of this technology toward spinal cord injury and disease. PMID- 11939462 TI - Relationship of respiratory symptoms with smoking status and pulmonary function in chronic spinal cord injury. AB - OBJECTIVE: The relationship of respiratory symptoms to pulmonary function parameters and smoking status was assessed in subjects with chronic (>1 year) spinal cord injury (SCI). METHODS AND PARTICIPANTS: As part of their annual physical examination, subjects were queried regarding respiratory symptoms and underwent pulmonary function studies. The 180 patients who successfully completed pulmonary function testing were evaluated, including 79 subjects with tetraplegia (56 nonsmokers and 23 smokers) and 101 subjects with paraplegia (78 nonsmokers and 23 smokers). FINDINGS: Logistic-regression analysis revealed the following independent predictors of breathlessness: level of injury (tetraplegia, paraplegia, odds ratio = 3.5, P < 0.0015), cough combined with phlegm and/or wheeze (CPWZ, odds ratio = 3.1, P < 0.015), total lung capacity percentage predicted (TLC <60%, odds ratio = 3.9, P < 0.02), and expiratory reserve volume (ERV < 0.6 L, odds ratio = 2.5, P < 0.05). Independent predictors of CPWZ were current smoking (odds ratio = 3.3, P < 0.004), breathlessness (odds ratio = 2.9, P < 0.03), and forced expiratory volume in 1 second (FEV1 <60%, odds ratio = 3.2, P < 0.01). CONCLUSION: Altered respiratory mechanics associated with tetraplegia contribute to breathlessness, restrictive ventilatory impairment (low TLC%), and reduced expiratory muscle strength (low ERV). These factors apparently overshadow adverse effects caused by smoking. Conversely, smoking and reduction of airflow (low FEV1%) were predictive of CPWZ, symptoms commonly associated with cigarette use. PMID- 11939463 TI - Spinal stenosis vs traumatic spinal cord injury: a rehabilitation outcome comparison. AB - OBJECTIVE: Nontraumatic spinal cord injury (NT/SCI), which can occur secondary to spinal stenosis, has been shown to represent a significant proportion of individuals admitted for SCI rehabilitation. The objective of this study was to compare demographics and outcomes of patients with spinal stenosis-induced SCI (SS/SCI) with those with traumatic spinal cord injury (T/SCI) following inpatient rehabilitation. DESIGN: This 7-year prospective review compared 81 patients with SS/SCI and 102 patients with T/SCI admitted to an SCI rehabilitation unit with similar levels and completeness of injury. Main outcome measures included rehabilitation hospital length of stay (LOS), Functional Independence Measure (FIM) scores, FIM change, FIM efficiency, rehabilitation charges, and discharge rates to home. RESULTS: Results indicate that, when compared with the T/SCI, patients with SS/SCI had a significantly (P < .05) higher mean age (64.1 years vs 44.4 years), were more often female (39% vs 20%), and tended to present with paraplegia vs tetraplegia (69% vs 46%) and with motor incomplete SCI vs incomplete SCI (100% vs 49%). When comparing etiologies of SCI within tetraplegic and paraplegic groups, results showed that individuals with tetraplegic SS/SCI had a significantly (P < .05) shorter rehabilitation LOS (25.7 vs 35.9 days), and lower FIM change (24.5 vs 32.5) and FIM efficiency (1.0 vs 1.3); however, no statistical differences were noted for discharge FIM scores and discharge to home rates. Individuals with paraplegic SS/SCI also had significantly lower FIM change (20.2 vs 28.7); however no significant differences were noted for rehabilitation LOS, charges, FIM efficiency, or discharge-to-home rates. CONCLUSIONS: The findings indicate that patients with SS/SCI present with less severe clinical impairments (motor incomplete and paraplegia) in comparison with patients with T/SCI. Clinically similar SS/SCI groups were noted to achieve rates of functional gain and community discharge comparable with T/SCI patients. Although patients with T/SCI achieved greater overall functional improvement, patients with SS/SCI had shorter rehabilitation LOS and lower rehabilitation charges. These findings have important implications for the interdisciplinary rehabilitation process in the overall management and outcome of individuals with NT/SCI. PMID- 11939464 TI - Early repolarization in patients with spinal cord injury: prevalence and clinical significance. AB - OBJECTIVES: The objective was to examine the prevalence of early repolarization in a spinal cord injury (SCI) clinic and the relationship of level of injury to this electrocardiogram (ECG) finding. BACKGROUND: ST elevation on the resting ECG can be either a normal variant or a sign of acute ischemia, evolving myocardial infarction, or pericarditis. It is frequently seen as a normal variant (early repolarization) in healthy individuals, but has also been reported in individuals with SCI. While the etiology of benign ST elevation (early repolarization) has not been clearly defined, current opinion is that this finding is seen in individuals with high vagal tone. METHODS: Retrospective analysis was made of 31 5 individuals with SCI at T5 or above (140 with complete injuries), and 1 98 with SCI at T6 or below, and who had ECGs in the computerized database at the Palo Alto VA Medical Center. A comparison cohort of 32,841 able-bodied male controls also was identified in the same ECG database. Patient demographics and computerized ST measurements were analyzed. RESULTS: The prevalence of ST elevation was significantly higher in both the total high-level injury group (19%) and the complete high-injury group (24.5%) than in either the low-injury (6.5%) or control groups (13%), with P < 0.001 for comparisons between both high- and low-injury groups and high injury vs control. The magnitude of ST elevation was also higher in the high-injury groups vs the low-injury and control groups. CONCLUSION: There is a higher prevalence of early repolarization in individuals with SCI at levels of injury that can disrupt central sympathetic command of the heart. It appears that either enhanced vagal tone or loss of sympathetic tone is responsible for ST elevation. PMID- 11939465 TI - Autonomic dysreflexia in multiple sclerosis. AB - BACKGROUND: Although autonomic dysreflexia (AD) is well documented in the spinal cord injury (SCI) population, its occurrence in persons with multiple sclerosis (MS) is not. A dense multiple sclerotic lesion in the spinal cord at or above the sixth thoracic level can cause interruption of descending inhibitory impulses and thus result in AD. A patient with MS presented to our facility with classic signs and symptoms of AD. We believe that lack of knowledge about the risks for this condition in MS led to a delay in diagnosis. METHODS: Case report illustrates AD in a person with MS. A convenience survey was conducted among clinicians who provide care to people with MS. The survey looked at both awareness of, and experience with, AD in MS. RESULTS: Forty-five percent of the respondents indicated they were not aware of the potential risk for AD among MS patients. Only 10% indicated they were aware of MS patients in their practice who had experienced AD. CONCLUSION: Although AD is probably less common in MS than in SCI, this case does not appear to be unique. Knowledge of this potential life threatening complication of MS seems to be limited. PMID- 11939466 TI - Gastric phytobezoar associated with impaired gastric motility in a patient with spinal cord injury. AB - Impaired gastrointestinal motility as a result of interruption of sympathetic outflow is a common occurrence in the spinal cord injury (SCI) population. In addition, frequent use of medications with anticholinergic properties in this population results in further impairment of peristalsis resulting in gastrointestinal stasis. Since SCI patients often lack sensation below the level of injury, they may present with vague symptoms, which complicates the diagnosis of intestinal obstruction. We report the first case of gastric phytobezoar in a patient with T4 ASIA A paraplegia who presented with vague upper abdominal discomfort, anorexia, weight loss, and vomiting. Because mortality rates can be as high as 30% if phytobezoars remain untreated, gastrointestinal phytobezoars should be considered in the differential diagnosis of abdominal discomfort in SCI patients. Etiologic factors for phytobezoars are discussed for the general population and in particular, for patients with SCI. PMID- 11939467 TI - Heterotopic ossification complicating prolonged intubation: case report and review of the literature. AB - BACKGROUND: Within the past decade several reports have been published concerning heterotopic ossification (HO) in adult respiratory distress syndrome patients subjected to prolonged mechanical ventilation. The knee has been the most common site of involvement, which tends to differentiate this entity of HO from those previously described. METHOD: Case report and literature review. FINDINGS: HO associated with prolonged intubation differs in clinical presentation from HO seen in spinal cord injury (SCI) and other trauma. Use of neuromuscular blockade does not appear to explain this risk. An unidentified humoral response mechanism may underlie the development of HO in these cases. Certain individuals may be genetically predisposed to develop HO. CONCLUSION: Increased awareness of this relatively new entity may assist early diagnosis, medical treatment, and eventually direct rehabilitation. Investigation of the pathogenesis of different types of HO may provide clues to the prevention and treatment of HO in individuals with SCI and other central nervous system trauma. PMID- 11939480 TI - Bovine spongiform encephalopathy: are the cows mad or full of carbohydrates? AB - The non-forage feeding of dairy cows rich in fast absorption carbohydrates, the low value of their euglycemic-hyperinsulinemic clamp suggest a dysregulation of carbohydrate metabolism able to produce neurodegenerative disorders. Comparisons between Alzheimer's disease developed in diabetes mellitus and bovine spongiform encephalopathy (BSE) direct the discussion of the origin of BSE not only towards a contamination by prion proteins. PMID- 11939481 TI - Plasma total antioxidant capacity in hemodialyzed patients and its relationships to other biomarkers of oxidative stress and lipid peroxidation. AB - Patients undergoing long-term hemodialysis (HD) exhibit increased levels of oxidative stress, likely contributing to the increased rate of cardiovascular disease. The present study represents a critical evaluation of some of the most widely used oxidative indicators, as applied to the monitoring of hemodialysis associated oxidative stress. Total plasma antioxidant capacity was determined by two independent procedures, the total antioxidant status (TAS) and the ferric reducing ability of plasma (FRAP) methods. Plasma lipid peroxidation was assessed by determining the peroxidation products malonaldehyde and 4-hydroxynonenal (MDA 4HNE) as well as lipid hydroperoxides ("Fox-2" and "d-ROMs" methods). Total plasma thiols and plasma alpha-tocopherol were also determined. MDA-4HNE levels were higher in HD patients and decreased following HD, possibly due to passive diffusion across dialysis filters. d-ROMs were also higher in HD patients but exhibited a further increase following the dialysis procedure. Serum alpha tocopherol did not show any significant differences. Plasma thiols were lower in HD patients and were restored following HD. Plasma total antioxidant capacity determined with either method was unexpectedly higher in HD patients compared to controls, and decreased following HD. These data indicate that, of the biomarkers studied, d-ROMs level is the one more accurately reflecting the oxidative alterations taking place in HD patients, while determination of MDA-4HNE fails to detect oxidation occurring during the HD sessions. In addition, our findings point out that the determination of total antioxidant capacity in HD patients is severely affected by the concomitant fluctuations in plasma urate levels and therefore needs careful interpretation. PMID- 11939482 TI - Effects of propofol and taurine on intracellular free amino acid profiles and immune function markers in neutrophils in vitro. AB - We have examined the effects of propofol, taurine, and the combination of propofol and taurine on amino acid profiles and the immune function markers superoxide anion (O2-), hydrogen peroxide (H2O2), and released myeloperoxidase (MPO) activity in neutrophils (PMN). Propofol led to significant changes in the dynamic PMN-free amino acid pool. Exogenous taurine significantly reduced PMN neutral amino acid and alpha-aminobutyrate (alpha-aba) as intracellular taurine increased. Incubation with propofol plus taurine resulted in lower intracellular taurine levels and elevated alpha-aba and neutral amino acid concentrations compared to propofol alone. Concerning PMN immune function markers, propofol significantly decreased O2- and H2O2 formation and released MPO. Taurine led to an increased release of MPO and concomitant significantly reduced O2- and H2O2 levels. When propofol and taurine were applied together they appeared to act additively with regard to superoxide and hydrogen peroxide formation. In the case of MPO, taurine neutralized propofol's effects, supporting the idea that MPO activity may be regulated by taurine. We believe therefore that taurine is important for strengthening PMN host defense capability, although the mechanisms are not yet clear. Moreover, taurine appears to act primarily by altering the PMN osmotic balance, while propofol seems to affect PMN amino acid metabolism and/or uptake and release. PMID- 11939483 TI - Genotyping of hemochromatosis-associated mutations in the HFE gene by PCR-RFLP and a novel reverse hybridization method. AB - Comparative analysis of the hemochromatosis-associated mutations C282Y, H63D and S65C in the HFE gene in 51 patients using three different methods is reported. One PCR-RFLP method was based on general primers, whereas another employed mutation-specific mismatched primers. The third method was a new PCR-based reverse hybridisation line probe assay (LiPA), comprising DNA amplification by general primers followed by a single step reverse hybridization to specific probes, immobilized on a nitrocellulose strip. Forty-eight (94%) of the 51 samples yielded identical results by all three methods. Three discrepant results were obtained, caused by polymorphisms in the primer binding region, resulting in no amplification at all or selective amplification, leading to misinterpretation of the HFE genotype by PCR-RFLP. The design of the assay and the stringency of the reaction conditions used are crucial to obtain a correct HFE genotype. PCR LiPA offers an easy and reliable alternative to currently used conventional methods. PMID- 11939484 TI - Comparison of different CD71 monoclonal antibodies for enrichment of fetal cells from maternal blood. AB - Different approaches have been proposed for the enrichment of fetal nucleated red blood cells (NRBC) from maternal blood as an alternative way to obtain fetal tissue for non-invasive prenatal diagnosis. The main purpose of this study was to compare two of our monoclonal antibodies (2E11.3 and 2B7.4 mAbs) with the most widely used commercial anti-CD71 mAb, in terms of their ability to isolate NRBC from maternal blood by magnetic activated cell sorting (MACS). Peripheral blood samples were obtained from 60 pregnant women at a mean gestational age of 16 weeks (range: 10-19 weeks). The number of NRBC isolated by our antibodies (median: 68, range: 0-2102) was significantly higher than that obtained by the commercial antibody (median: 38, range: 0-2165) in the same samples. However, in the final preparations, contamination by maternal nucleated blood cells was lower when the commercial antibody was used. Since fetal NRBC are rare in maternal blood, the improved NRBC recovery achieved by our non-commercial antibodies should facilitate the non-invasive detection of fetal aneuploidies in maternal blood. PMID- 11939485 TI - The effects of marathon swimming on serum leptin and plasma neuropeptide Y levels. AB - It seems likely that the neuropeptide Y (NPY)-leptin axis is involved in the regulation of energy expenditure in man. The purpose of this study was to observe the effect of a model of intense prolonged exercise-mediated energy expenditure (25 km swim race in 6.9-10.5 hours) on leptin and NPY concentrations in male long distance swimmers. Sixteen long-distance swimmers (mean age 25, range 18-45 years) who took part in a 25 km sea swimming competition (Toroneos golf, Chalkidiki, Greece) participated in the study. Mean competition time was 8.5 hours (range 6.5-10.5). The participants were allowed food and beverage intake ad libitum before and throughout the 25 km race. Venous blood samples were taken prior and immediately after the race for the measurement of serum leptin and plasma NPY. Non-esterified free fatty acids (NEFFA) and glycerol levels were determined as indicators of adipose tissue lipids mobilization. Results showed that leptin levels after marathon swimming were significantly reduced (p<0.001) in all athletes. There was a statistically significant negative correlation (r= 0.812, p<0.01) between the values of leptin and glycerol just after the termination of swimming. Blood serum glycerol and free fatty acid levels were significantly increased (p<0.001) in all swimmers. Plasma NPY levels were also increased (p<0.01) in 81.2% of the swimmers. Linear regression analysis revealed a significant negative correlation between the values of leptin and NPY (r= 0.789, p<0.01). In conclusion, these data support our initial hypothesis that appropriate changes in leptin and NPY take place during marathon swimming to compensate for the negative energy balance produced due to this prolonged effort. This indicates the NPY-leptin axis involvement in the regulation of energy expenditure in man. PMID- 11939486 TI - Renal function exerts only a minor influence on high plasma homocysteine concentrations in patients with acute coronary syndromes. AB - It has been suggested that hyperhomocysteinemia observed in patients with occlusive vascular disease is caused by reduced renal function secondary to renovascular disease. We have therefore used serum cystatin C, a new sensitive marker for glomerular filtration, in 59 patients with acute coronary syndromes and high plasma homocysteine (tHcy) concentration to measure renal function. Samples were also obtained from 34 patients with low-normal plasma tHcy and 50 control subjects. The patients with low-normal plasma tHcy concentration showed decreased concentrations of serum cystatin C and serum creatinine and increased concentrations of blood folate and serum cobalamin compared to the controls and to the patients with high plasma tHcy. There was a large overlap in cystatin C concentrations between patients with high and low-normal plasma tHcy. None of the parameters investigated except plasma tHcy were significantly different in the group of patients with high plasma tHcy concentration compared to the control group. In order to further demonstrate the importance of renal impairment, a subgroup of the patients with high plasma tHcy was supplemented daily with folic acid 5 mg, pyridoxine 40 mg and cyancobalamin 1 mg for 3 months. Vitamin therapy reduced plasma tHcy from 18.3+/-4.6 pmol/l to 9.6+/-2.2 pmol/l (p<0.0001). However, vitamin treatment did not strengthen the correlation between cystatin C and plasma tHcy concentrations. These findings do not support the hypothesis that subtle renal dysfunction is an important cause of high plasma tHcy concentration in patients with acute coronary syndromes. PMID- 11939487 TI - Changes of biochemical bone markers during the menopausal transition. AB - We present data on the changes of the bone formation markers osteocalcin (OC), bone-specific alkaline phosphatase (bone ALP) and bone sialoprotein (BSP), as well as the resorption markers pyridinoline (PYD), deoxypyridinoline (DPD), C- and N-terminal telopeptide cross-linked collagen type I (CTX, NTX), and tartrate resistant acid phosphatase type 5b (TRACP) at five time points during the course of two years in healthy premenopausal, perimenopausal and early postmenopausal women. The prospective study showed that CTX (p<0.001), NTX (p=0.001) and TRACP (p=0.001), as well as bone ALP (p=0.009) and OC (p=0.052), were significantly increased already in the transition period from peri- to postmenopause. The pyridinium crosslinks indicated an increased collagen degradation rate already in the perimenopause (PYD, p=0.017; DPD, p=0.054). Significant inverse correlations with the two years changes of the bone mineral density were found for bone ALP, CTX, OC and DPD in the perimenopausal group. The measurement of a comprehensive panel of biochemical bone markers clearly shows that metabolic changes in bone metabolism appear pronounced in the perimenopause, a period still presenting satisfactory estrogen supply. Thus, the perimenopause is an important phase for a contingent development of osteoporosis. PMID- 11939488 TI - Blood sampling for biochemical analysis from central venous catheters: minimizing the volume of discarded blood. AB - The purpose of the work was to establish a reliable procedure for blood sampling from central venous catheters with a minimal loss of blood. Based on previous studies of potassium measurements, the routine procedure had been to discard 12 ml of blood. In order to clear the catheter from substances from the infusion including potassium, the catheters were first flushed with 5 ml of saline. Repeated samples in fractions of 3 ml for hematological analyses (ethylenediaminetetraacetic acid plasma; seven patients), coagulation tests (citrate plasma; seven patients) and for serum measurements (eight patients) showed that a discarded volume of 3 ml corresponding to at least six times the dead-space of the catheters was sufficient after initial flushing with saline. PMID- 11939489 TI - Experience with external quality assessment of drugs of abuse testing in the Lombardy Region in Italy. AB - Following up previous experience with External Quality Assessment (EQA) and Proficiency Testing Programs (PTP) on drugs of abuse (DoA) testing in Italy and in other European countries, the government of the Lombardy Region, first among Italian regions, established in 1995 a compulsory EQA scheme for laboratories authorized to perform these tests. The purpose of the present work is the description of the program and the overall evaluation of the results obtained in the first three annual cycles (1995-1998). During each annual cycle laboratories received 22 urine samples; some samples were collected from patients ("real samples") and some were "spiked" urine samples. Both types of samples could contain the following substances/classes of substances: opiates, cocaine, cannabinoids, methadone, buprenorphine, benzodiazepines, barbiturates. Type A laboratories used an immunological screening method; they expressed the results as concentrations. Type B laboratories, authorized to perform screening methods followed by confirmatory techniques, searched for and identified single substances and provided an interpretation on possibly taken drugs. During the study period the laboratories produced about 21,000 analytical results. Among them, false-negative results were 0.9% of true positives, and false-positive results were 0.7% of true negatives. Performance using the spiked samples was better than using real samples, and performance of type B laboratories better than that of type A. The results obtained during the program are consistent with those of other quality control programs. This program, in addition, has provided information on the status of DoA testing in the Lombardy region's laboratories, in particular on their analytical performance, on the quality of interpretation of results and on a degree of improvement achieved during the program. PMID- 11939490 TI - Reference intervals for thyroid hormones on the architect analyser. AB - The objective of this study was to establish reference intervals for thyroid stimulating hormone (TSH), free thyroxine (FT4), free triiodothyronine (FT3), total thyronine (TT4) and total triiodothyronine (TT3) on the Architect i2000 analyser (Abbott). Serum samples were obtained from apparently healthy adults (n=217, age 18-90 years) excluding individuals taking oral contraceptives or under hormone replacement therapy. The second group were ambulatory euthyroid patients (n=323) excluding those with a history of thyroid disorders. We also investigated thyroid hormones in sera from euthyroid hospitalised patients (n=490) excluding those with severe non-thyroidal illness. The reference intervals for the healthy adults were for TSH 0.17-4.23 mIU/l, for FT4 11.24 26.86 pmol/l, for FT3 2.56-6.36 pmol/l, for TT4 55.8-155.1 nmol/l and for TT3 0.90-2.54 nmol/l. TSH and TT3 concentrations were similar in males and females. However, FT4, FT3 and TT4 levels exhibited significant differences between females and males. No significant differences were observed between the concentrations of TSH, FT3, TT3, FT4 and TT4 in healthy subjects and in euthyroid ambulatory patients aged 18-90 years. TSH levels in healthy subjects were the same in younger and older individuals. In contrast, in outpatients and in hospitalised patients TSH concentrations were significantly lower (20%) in subjects older than 50 years compared to those younger than 50 years. For FT3 and TT3 we consistently observed in all three study groups 6-7% and 8-12% higher concentrations in the younger (< 50 years) compared to the older (> 50 years) subjects. For FT4 and TT4 no consistent pattern of correlation with age was detectable when the three study groups were analysed independently. The reference intervals for thyroid hormones determined in this study differ considerably from values found in other European and non-European countries. This underlines the need for population-specific reference ranges. PMID- 11939491 TI - Intermethod variation in serum carcinoembryonic antigen (CEA) measurement. fresh serum pools and control materials compared. AB - This study was undertaken to evaluate the feasibility of using commercial control materials in a regional external quality assessment scheme (EQAS) for serum carcinoembryonic antigen (CEA) measurement. We have assessed the commutability of 12 commercial control materials using five automated immunochemical systems. We compared the intermethod behavior of the materials with that of 12-14 patient serum pools. In a total of 48 comparisons (12 materials x 4 pairs of analytical systems), seven instances of non-commutability were apparent, as shown by normalized residuals falling outside the +/-3 interval. The use of non-commutable materials generates two negative effects. In EQAS, the interlaboratory variation recorded is not representative of the variation expected in the assay of patient sera; in interlaboratory harmonization programs, recalibration with non commutable materials increases, instead of decreasing, the interlaboratory variation. Both these effects were shown to occur in CEA measurement with the tested materials and systems. The materials planned to be used in our EQAS turned out to be commutable: this gave us the guarantee of measuring realistic interlaboratory variation values, although the check for commutability should be extended to all the analytical systems used by the participants in the scheme. PMID- 11939492 TI - Turn-around time for chemical and endocrinology analyzers studied using simulation. AB - Simulation can be a means for the laboratory management to investigate the effects of proposed changes in the laboratory. Also, when specifications of analyzers are available one can investigate which analyzer should be purchased to fulfill existing needs. The potential of simulation is shown here by simulating the turn-around time of batched samples under several conditions. Three routine analyzers (DAX, AXON and Immuno-1) were modeled. The temporal pattern of the arrival of the samples at the different analyzers was determined. The impact of these patterns and the two batching methods on the turn-around time was investigated. Simulation proved to be a useful method to study the effects of batching and arrival patterns on the turn-around time. PMID- 11939493 TI - A two-center evaluation of the blood gas immediate response mobile analyzer (IRMA). AB - The Immediate Response Mobile Analyzer (IRMA) is a selective and portable point of-care testing (POCT) blood gas, electrolyte and hematocrit (Hct) analyzer. The overall analytical performance was evaluated in a two-center study involving two Italian hospital laboratories, following the guidelines suggested by the manufacturer (based on the NCCLS protocol), after a preliminary evaluation of their formal validity. The IRMA was compared to the analyzers used in the routine laboratory as reference. The considered parameters were pH, pO2, pCO2, Na+, K+, ionized calcium and Hct. When using the aqueous quality control material provided by the manufacturer most of the parameters showed good precision, with the exception of pCO2 and pO2 that showed high CVs on two of the three levels of the aqueous control. We could demonstrate that this imprecision was material-related and was reduced when using a different material (blood equilibrated by tonometry). With tonometred blood for pO2 and pCO2 and the aqueous material for the remaining parameters the CVs were all below 5%, ranging from 0.08% to 2.8%. The IRMA results correlated adequately with the comparison instruments, with the exception of sodium and ionized calcium where contradictory results were obtained in the two centers. PMID- 11939494 TI - Painting, poetry and optics: Johannes Vermeer. AB - The painter Johannes Vermeer (1632-75) worked in the town of Delft in the Dutch Republic. This article focuses on associations between his painting method and the expertise in optics available in Delft at the time. His art represents an extraordinary combination of technical prowess and poetic expression. PMID- 11939495 TI - The practice of clinical chemistry in the European Union. AB - The European Communities Confederation of Clinical Chemistry has been actively engaged in raising the level of clinical chemistry in the European Union. Closer contacts between the national societies for clinical chemistry have resulted in more comparable programs for postgraduate training of clinical chemists, closer similarity of contents and practice of the profession in the different countries, and the official registration of professionals. This article reviews some of the characteristics of professional organisation, practice, and regulation in the fifteen European Union countries. Many similarities appear. In half of the countries microbiology, blood-banking and transfusion medicine fall within the domain of clinical chemistry. The minimum number of years for training (university and postgraduate) is eight, but in practice this will extend to 10 or more years. Official regulation of the profession by law exists in a minority of countries. Continuing education and re-registration have not been officially instituted yet in any country, but these issues will be the next steps forward. In those countries that prepare themselves for entering the European Union, training and practice of clinical chemistry are moving towards the common standards of the European Communities Confederation of Clinical Chemistry. PMID- 11939496 TI - Recommendations for the routine use of pancreatic amylase measurement instead of total amylase for the diagnosis and monitoring of pancreatic pathology. AB - This document reviews the scientific evidence expected to persuade clinical laboratories to substitute pancreatic amylase measurement for total amylase in cases of suspected pancreatic pathology. A substantial evidence is now available to support such change. The measurement of pancreatic amylase in serum is: 1. more sensitive and specific for the detection of pancreatic tissue damage than that of the total enzyme activity, 2. easy and quick to perform in emergency conditions, 3. analytically precise in relation to its clinical application, 4. suitable for easy transfer and comparison of results from different care delivery units, and 5. characterized by well-defined decision limits for the diagnosis of acute pancreatitis. PMID- 11939497 TI - Hypothermia on admission in patients with severe brain injury. AB - Data from the "National Acute Brain Injury Study: Hypothermia" were examined to identify the impact of hypothermia on admission. In all patients, temperature was measured at randomization using bladder catheters with thermistors. Patients assigned to hypothermia were cooled using fluid-circulating pads. Outcome was assessed at 6 months using the dichotomized Glasgow Outcome Scale (good outcome = good recovery/moderate disability; poor outcome = severe disability/vegetative/dead). One-hundred and two patients (hypothermia, 62; normothermia, 40) were hypothermic on admission (< or =35.0 degrees C). Hypothermia-on-admission patients assigned to normothermia (n = 40) had a 78% poor outcome, and normothermia-on-admission patients assigned to normothermia had a 52% poor outcome (p < 0.004). Hypothermia-on-admission patients assigned to hypothermia had a lower percentage of poor outcomes than those assigned to normothermia (hypothermia, 61%; normothermia, 78%; p = 0.09). Patients over 45 years of age had an adverse effect of hypothermia regardless of admission temperature due to medical complications. Patients who were hypothermic on admission, age < or = 45 years (n = 81), and assigned to hypothermia had a significantly lower percentage of poor outcomes than those assigned to normothermia (hypothermia, 52%; normothermia, 76%; p = 0.02). Factors associated with hypothermia on admission were increased age, prehospital hypotension, smaller size, positive blood alcohol, larger volume of pre-hospital fluids, slightly higher injury severity, and winter enrollment The treatment effect was found in all of the four centers, which randomized the majority (80%) of the patients. It is unclear whether the improved outcome when hypothermia is maintained is a beneficial effect of very early hypothermia induction or an adverse effect of permitting the patients to rewarm passively. PMID- 11939498 TI - Craniectomy position affects morris water maze performance and hippocampal cell loss after parasagittal fluid percussion. AB - Valid and reliable animal models are essential for mechanistic and therapeutic studies of traumatic brain injury (TBI). Therefore, model characterization is a continual and reciprocal process between the experimental laboratory and the clinic. Several excellent experimental models of TBI, including the lateral fluid percussion rat model, are currently in wide use in many neurotrauma laboratories. However, small differences in the position of lateral fluid percussion craniectomy are reported between labs. Additionally, differences in hippocampal cell death have also been reported. Therefore, we hypothesized that small changes in craniectomy position could affect commonly used outcome measures such as vestibulomotor function, Morris water maze (MWM) performance, hippocampal cell loss, and glial fibrillary acidic protein (GFAP) immunoreactivity. Four placements were systematically manipulated: rostral, caudal, medial, and lateral. The medial and caudal placements produced significantly greater impairments in the MWM acquisition task over the lateral and rostral placements. The rostral placement produced diffuse cortical damage but little hippocampal cell loss. In contrast, the medial, lateral, and caudal placements produced more mid-dorsally localized cortical damage and significant cell loss in the CA2/CA3 and hilus ipsilateral to the injury site. Furthermore, reactive astrocytosis was more pronounced in the medial, lateral, and caudal placements than in the rostral placement. All craniectomy position groups had similar durations of traumatic unconsciousness and similar impairment on motor tasks. We conclude that small alterations in craniectomy position produce differences in cognitive performance, hippocampal cell loss, and reactive astrocytosis but not in motor performance nor transient unconsciousness. PMID- 11939499 TI - Acute ethanol intake attenuates inflammatory cytokines after brain injury in rats: a possible role for corticosterone. AB - It has been reported that acute ethanol intoxication exerts dose-dependent effects, both beneficial and detrimental, on the outcome of traumatic brain injury (TBI), although the mechanism(s) has not been determined. Given that pro inflammatory cytokines are either neuroprotective or neurotoxic, depending on their tissue levels, ethanol-induced alterations in brain cytokine production may be involved in determining the recovery after TBI. The present study was undertaken to examine the effect of acute ethanol pretreatments (producing blood alcohol concentrations of 100+/-16 mg/dL, and 220+/-10 mg/dL, considered low and intoxicating doses, respectively) on interleukin-1beta (IL-1beta) and tumor necrosis factor-alpha (TNF-alpha) levels in discrete brain regions. In addition, serum corticosterone levels were also examined because the hormone is a modulator of cytokine production, its secretion is stimulated by ethanol, and it has been associated with the severity of post-injury neurologic dysfunction. The data presented in this report demonstrate that moderate cortical impact brain injury elicits a marked increase in IL-1beta and TNF-alpha in the injured cortex as well as in the hippocampus ipsilateral to the injury. Ethanol pretreatment lowered cytokine levels in the cortex, hippocampus and hypothalamus in a dose-dependent manner after TBI compared to the untreated injured rats. Serum corticosterone levels were markedly increased in the injured rats, and were further augmented in the ethanol-pretreated injured animals in a dose-dependent manner. Our findings suggest that ethanol-induced decrease in pro-inflammatory cytokine production may be linked to increased circulating corticosterone, both of which may contribute to the outcome of brain injury. PMID- 11939500 TI - The effects of L-arginine on cerebral hemodynamics after controlled cortical impact injury in the mouse. AB - Traumatic brain injury (TBI) induces vascular changes that may influence neurological outcome by causing the brain to be more susceptible to secondary ischemic insults. In rat models of TBI, L-arginine administration has been shown to restore cerebral blood flow and improve neurological outcome. The purpose of this study was to determine if hypoperfusion occurs in a mouse model of TBI and if L-arginine administration has the same beneficial effects after injury in the mouse. C57BL6 mice were anesthetized with isoflurane, intubated and mechanically ventilated, and underwent a 3-m/sec, 1.5-mm deformation cortical impact injury. Five minutes after injury, L-arginine, 300 mg/kg, or saline were administered. Arterial blood pressure, intracranial pressure, and laser Doppler flow at the impact site were monitored for 3 h after the injury. The cerebral hemodynamic effects of the TBI induced by cortical impact injury were similar to that previously observed in rats. Intracranial hypertension, with ICP peaking at 46+/ 2 mm Hg, and systemic hypotension both contributed to a reduction in CPP. In addition, LDF decreased significantly at the impact site. L-Arginine administration restored LDF to near baseline levels without increasing ICP. These studies demonstrate that cerebral hemodynamics can be measured in mouse models of TBI. The changes in cerebral hemodynamics are relatively simlar to those see in the rat model of cortical impact injury and suggest an important role for nitric oxide metabolism in the maintenance of cerebral blood flow following TBI. PMID- 11939501 TI - Role of RyRs and IP3 receptors after traumatic injury to spinal cord white matter. AB - Calcium influx and elevation of intracellular free calcium (Ca2+i), with subsequent activation of degenerative enzymes is hypothesized to cause cell injury and death after trauma. We examined the effects of traumatic compressive injury on (Ca2+)i dynamics in spinal cord white matter. We conducted electrophysiological studies with ryanodine and inositol (1,4,5)-triphosphate (IP3) receptor agonists and antagonists in an in vitro model of spinal cord injury (SCI). A 25-30-mm length of dorsal column was isolated from the spinal cord of adult rats, pinned in an in vitro recording chamber (37 degrees C) and injured with a modified clip (2-g closing force) for 15 sec. The functional integrity of the dorsal column was monitored electrophysiologically by quantitatively measuring the compound action potential (CAP) with glass microelectrodes. The CAP decreased to 55.2+/-6.8% of control (p < 0.05) after spinal cord injury (SCI). Chelation of Ca2+i with BAPTA-AM (a high-affinity calcium chelator) promoted significantly greater recovery of CAP amplitude (83.2+/-4.2% of control; p < 0.05) after injury. Infusion of caffeine (1 and 10 mM) exacerbated CAP amplitude decline (45.1+/-5.9% of control; p < 0.05; 44.6+/ 3.1% of control; p < 0.05) postinjury. Blockade of Ca2+i release through ryanodine-sensitive receptors (RyRs) with dantrolene (10 microM) and ryanodine (50 microM), conferred significant (p < 0.05) improvement in CAP amplitude after injury. On the other hand, blockade of Ca2+i with inositol (1,4,5)-triphosphate receptor (IP3Rs) blocker 2APB (10 microM) also conferred significant improvement in CAP amplitude after injury (82.9+/-7.9%; p < 0.05). In conclusion, the injurious effects of Ca2+i in traumatic central nervous system (CNS) white matter injury appear to be mediated both by RyRs and through IP3Rs calcium-induced calcium release receptors (CICRs). PMID- 11939502 TI - Cellular basis for progesterone neuroprotection in the injured spinal cord. AB - Progesterone (PROG) exerts beneficial and neuroprotective effects in the injured central and peripheral nervous system. In the present work, we examine PROG effects on three measures of neuronal function under negative regulation (choline acetyltransferase [ChAT] and Na,K-ATPase) or stimulated (growth-associated protein [GAP-43]) after acute spinal cord transection injury in rats. As expected, spinal cord injury reduced ChAT immunostaining intensity of ventral horn neurons. A 3-day course of intensive PROG treatment of transected rats restored ChAT immunoreactivity, as assessed by frequency histograms that recorded shifts from predominantly light neuronal staining to medium, dark or intense staining typical of control rats. Transection also reduced the expression of the mRNA for the alpha3 catalytic and beta1 regulatory subunits of neuronal Na,K ATPase, whereas PROG treatment restored both subunit mRNA to normal levels. Additionally, the upregulation observed for GAP-43 mRNA in ventral horn neurons in spinal cord-transected rats, was further enhanced by PROG administration. In no case did PROG modify ChAT immunoreactivity, Na,K-ATPase subunit mRNA or GAP-43 mRNA in control, sham-operated rats. Further, the PROG-mediated effects on these three markers were observed in large, presumably Lamina IX motoneurons, as well as in smaller neurons measuring approximately <500 micro2. Overall, the stimulatory effects of PROG on ChAT appears to replenish acetylcholine, with its stimulatory effects on Na,K-ATPase seems capable of restoring membrane potential, ion transport and nutrient uptake. PROG effects on GAP-43 also appear to accelerate reparative responses to injury. As the cellular basis for PROG neuroprotection becomes better understood it may prove of therapeutic benefit to spinal cord injury patients. PMID- 11939503 TI - AFGF promotes axonal growth in rat spinal cord organotypic slice co-cultures. AB - This study developed a slice culture model system to study axonal regeneration after spinal cord injury. This model was tested in studies of the roles of acidic fibroblast growth factor (aFGF) and peripheral nerve segments in axonal growth between pieces of spinal cord. Transverse sections of P15-P18 Sprague-Dawley rat spinal cord were collected for organotypic slice cultures. Group I consisted of two slices of spinal cord in contact with each other during the culture period. Group II consisted of two slices that were separated by 3 mm and connected by two segments of intercostal nerves. Group III consisted of single slices for studies of neuron survival. Some cultures from each group included aFGF in the culture medium. Bromodeoxyuridine (BrdU) was included in the medium for some cultures. The results showed three principal findings. First, counts of neurofilament positive cells demonstrated that treatment with aFGF significantly increased the number of surviving neurons in culture. Second, neurofilament immunostaining and DiI tracing demonstrated axons crossing the junction between the two pieces of spinal cord or growing through the intercostal nerve segments, and these axons were seen only in cultures with aFGF treatment. Third, few cells were double stained for neurofilament and BrdU, and these were found only with aFGF treatment. These results demonstrate that (1) organotypic slice cultures present a useful model to study regeneration from spinal cord injury, (2) aFGF rescues neurons and promotes axonal growth in these cultures, and (3) segments of intercostal nerves promote axon growth between slices of spinal cord. PMID- 11939504 TI - Enhanced survival and regeneration of axotomized retinal ganglion cells by a mixture of herbal extracts. AB - The aim of this study is to investigate the effects of Panax quinquefolius L. extract (PQE), Ginkgo biloba extract (GBE), and Hypericum perforatum extract (HPE), in combination or alone, on the survival and regeneration of axotomized retinal ganglion cells (RGCs) in an optic nerve transection model in adult hamsters. Unilateral transection of the optic nerve was performed to evaluate the effects of herbal extracts on the survival of axotomized RGCs. Effects of the herbal extracts on axonal regeneration of axotomized RGCs, on the other hand, were studied by attaching a peripheral nerve graft onto the transected ocular stump to induce regeneration. Operated animals received daily oral administration of vehicle or herbal extracts (PQE, GBE, and HPE), alone or in combination, for 7 and 21 days, respectively, in the survival and regeneration experiments. Surviving and regenerating RGCs were retrogradely labeled with Fluoro-Gold. The eyes were then enucleated and the retinas were flat-mounted for the counting of the labeled RGCs. Treatment with PQE, GBE and HPE alone failed to offer neuroprotection to injured RGCs. However, treatment with Menta-FX, a mixture of PQE, GBE, and HPE, significantly augmented RGC survival 7 days postaxotomy. Treatment with Menta-FX also induced a significant (87%) increase in the number of regenerating RGCs 21 days after optic nerve transection. This study demonstrates that herbs can act as a potential neuroprotective agent for damaged RGCs. It also suggests that the therapeutic value of herbal remedies can be maximized by the use of mixtures of appropriate herbs. PMID- 11939505 TI - Expression of c-Fos and c-Myc and deposition of beta-APP in neurons in the adult rat brain as a result of exposure to short-lasting impulse noise. AB - There is increasing evidence that impulse noise causes brain damage, but little is known about the mechanisms and extent of the response. Here, rat brains were investigated immunohistochemically for the expression of c-Fos, c-Myc, and beta APP during the first 3 weeks postexposure to impulse noise of 198 or 202 dB. The expression of c-Fos and c-Myc increased at 2 h after exposure in neurons of the cerebral cortex, thalamus, and hippocampus, and this c-Fos immunoreactivity remained elevated for the entire observation period. The c-Myc immunoreactivity peaked at 18 h in both neurons and astrocytes but returned to control levels at 7 days. Abnormal deposition of beta-APP was evident within 6 h in the same brain regions. The beta-APP immunoreactivity was most prominent at 18 h and remained increased over the 21-day period assessed. The observed effects were similar to those described in humans following traumatic brain injury and in Alzheimer's disease. We conclude that impulse noise influences the brain in a fashion similar to that in cases with progressive CNS degeneration. PMID- 11939506 TI - Hb A2-Monreale [delta146(HC3)His-->Arg], a novel delta chain variant detected in west Sicily. AB - We report a novel mutation, Hb A2-Monreale [delta146(HC3)His-->Arg], detected by cation exchange high performance liquid chromatography in a family from West Sicily. The mutation is due to a CAT-->CGT substitution at codon 146 of the delta globin gene. The two carriers had reduced levels of normal Hb A2 (1.1%), but comparable levels (0.9%) of the Hb A2 variant. Most likely the new variant has the same characteristics as Hb Cochin-Port Royal [beta146(HC3)His-->Arg], that is stable but has a 75% reduction of the Bohr effect. The finding of the new variant increases the genotype heterogeneity of the delta-globin gene in the Mediterranean area, and is relevant to the study and prevention of Cooley's Anemia. PMID- 11939507 TI - Prevalence of -alpha3.7-thalassemia in Argentina. PMID- 11939508 TI - Hb O-Tibesti [beta121(GH4)Glu-->Lys; beta11(A8)Val-->Ile], a hemoglobin variant carrying in the same beta chain the substitutions of Hb O-Arab and Hb Hamilton, found in combination with Hb S [beta6(A3)Glu-->Val]. AB - Hb O-Tibesti, carries in the same chain the substitution of Hb O-Arab [beta121(GH4)Glu-->Lys] and that of Hb Hamilton [beta11(A8)Val-->Ile]. Hb O Tibesti may be distinguished from Hb O-Arab by polyacrylamide gel electrophoresis in the presence of urea and Triton-X100, and by reversed phase high performance liquid chromatography. It was found in a compound heterozygous condition with Hb S [beta6(A3)Glu-->Val] in a child of Chad-Sudanese descent, suffering from a sickle cell syndrome. Compared to the classical description of the Hb S/Hb O-Arab association, the additional Hb Hamilton mutation does not seem to modify the clinical presentation. PMID- 11939509 TI - Hb Vila Real [beta36(C2)Pro-->His] in Italy: characterization of the amino acid substitution and the DNA mutation. AB - A rare high oxygen affinity hemoglobin variant was identified in a 22-year-old male patient from Napoli (Naples, Italy) affected by erythrocytosis. A detailed structural characterization of the variant hemoglobin was carried out, both at the protein and DNA levels essentially by mass spectrometric procedures and allele-specific amplification techniques. The amino acid substitution was determined by liquid chromatography tandem mass spectrometric analysis of the tryptic digest as beta36(C2)Pro-->His; the corresponding DNA mutation was identified as C-->A at the second position of codon 36 of the beta chain (CCT- >CAT). These variations identified the presence of Hb Vila Real, described only once before in a Portuguese woman. Haplotype analysis of DNA polymorphisms showed that the beta-globin gene of Hb Vila Real was associated with haplotype I. PMID- 11939510 TI - The beta+-IVS-I-6 (T-->C) mutation accounts for half of the thalassemia chromosomes in the Palestinian populations of the mountain regions. AB - A study of the spectrum of beta-thalassemia mutations in the southern part of the West Bank of the Palestinian Authority revealed the presence of 10 different beta globin mutations. The study included 41 patients and 54 carriers of beta thalassemia and sickle cell anemia. The spectrum of mutations observed was typically Mediterranean. However, their relative frequencies was unique. The predominant allele was IVS-I-6 (T-->C), with an exceptionally high frequency of 48.5% for this mutation. The homozygous IVS-I-6 patients had widely variable clinical presentations, from typical transfusion-dependent thalassemia major to non-transfusion-dependent thalassemia intermedia phenotype. Since it is so widespread in these West Bank populations, the IVS-I-6 mutation may date back to ancient times. The nonsense mutation at codon 37 (G-->A) was found at a relatively high frequency of 11.3%, supporting the hypothesis that it originated in this region. The other mutations, at decreasing frequencies ranging from 9.5 1.5%, were: IVS-I-110 (G-->A), frameshift codon 5 (- CT), IVS-I-1 (G-->A), IVS-II 1 (G-->A), Hb S [beta6(A3)Glu-->Val], frameshift codons 8/9 (+G), codon 39 (C- >T), and -30 (T-->A). Our findings will improve health care for the Palestinian population, and also has implications for the study of the origin and spread of thalassemia in the Middle East. PMID- 11939511 TI - A novel beta0-thalassemia mutation at codon 55 (-A) and a rare 17 bp deletion at codons 126-131 in the Indian population. AB - A new mutation at codon 55 (-A) and a rare mutation, a 17 bp deletion at codons 126-131, that gives rise to beta0-thalassemia, were found in the Indian population by means of direct sequencing of two polymerase chain reaction products generated from a 2.3 kb DNA fragment containing the whole beta-globin gene. Each polymerase chain reaction product was sequenced on both strands in a mutation-loading format, showing all nucleotide substitutions or deletions/insertions, including mutations and polymorphisms, in the product. The entire protocol requires four sequencing reactions/gel loadings after two successive polymerase chain reactions, which simplifies the mutation search process and reduces the reading error rate. PMID- 11939512 TI - A novel beta-thalassemia mutation in an Asian Indian. AB - A novel 7 bp deletion in exon 2 of the beta-globin gene in a 9-year-old boy originating from the eastern part of India is described. This deletion causes a shift in the reading frame of the beta-globin coding sequences, and consequently, a premature translation termination due to the creation of a stop codon at position 86. A slipped strand mispairing during DNA replication repair is proposed as the potential mechanism in generating this small deletion. PMID- 11939513 TI - Hb G-San Jose variant levels correlate with alpha-thalassemia genotypes. AB - Hb G-San Jose or beta7(A4)Glu-->Gly has been reported in Southern Italian or Mexican families. We have studied four families from Sicily and Campania, Southern Italy. In six carriers, the hemoglobin variant level ranged from 32 to 38%. In four double heterozygotes for Hb G-San Jose and alpha-thalassemia the variant level showed a strong correlation with the alpha-thalassemia genotype. In fact, the variant level was 15% when interacting with the - (alpha)20.5/alphaalpha, 19.6% with the alphaalpha/alphaPoly Aalpha, and 24.8% with alphaalpha/alpha(-5) ntalpha genotypes. In two double heterozygotes for Hb G San Jose and beta+ -IVS-I-6 (T-->C) the hemoglobin variant level was 67%. These data show that the reduced synthesis of alpha chains causes drastic reduction of probability to form Hb G-San Jose in favor of the formation of Hb A. Moreover, this reduction, (i) correlates with the type of alpha-thalassemia genotype and with the degree of the alpha chain deficiency, and (ii) is, most probably, more marked than the degree of alpha chain reduction. The minor affinity of the beta chain variant for the alpha chains associated with the reduced synthesis of the alpha chains is probably the principal cause of the variant hemoglobin reduction. Moreover, the rapid removal of the abnormal chains by proteolytic enzymes must have an essential role in order to reduce the chain variant pool. These conclusions are in agreement with the results obtained in reticulocyte and in vitro recombination experiments. PMID- 11939514 TI - Hb Canterbury [beta112(G14)Cys-->Phe]: a new, mildly unstable variant. PMID- 11939515 TI - Hb Molfetta [beta126(H4)Val-->Leu, GTG-->CTG]: a new, silent, neutral beta chain variant found in an Italian woman. AB - We have identified a new neutral hemoglobin variant in a pregnant Italian woman, that resulted from a GTG-->CTG replacement at codon 126 of the beta chain, corresponding to a Val-->Leu amino acid change at position beta126(H4). Thermal and isopropanol stability tests were normal and there were no abnormal clinical features. Routine electrophoretic and ion exchange chromatographic methods for hemoglobin separation failed to show this variant, but reversed phase high performance liquid chromatography revealed an abnormal peak eluting near the normal beta chain. No abnormal tryptic peptide was revealed on the high performance liquid chromatographic elution pattern of the total globin digest. The mutation was determined at the DNA level by amplification of the three beta exons by polymerase chain reaction and direct sequencing of one exon that showed an abnormal migration on single strand conformational polymorphism analysis. PMID- 11939516 TI - Identification of Hb Kenya (Agamma81Leu-beta86Ala) by electrospray mass spectrometry. PMID- 11939517 TI - Hb Siam [alpha15(A13)Gly-->Arg (alpha1) (GGT-->CGT)] is a typical alpha chain hemoglobinopathy without an alpha-thalassemic effect. PMID- 11939518 TI - Dominant beta-thalassemia due to a newly identified frameshift mutation in exon 3 (codon 113, GTG-->Tg). PMID- 11939519 TI - Identification of two new beta-thalassemia splice mutations: IVS-I-1 (G-->C) and IVS-I (-2) (A-->C). PMID- 11939521 TI - Hb G-Chinese: a G-->C substitution at codon 30 of the alpha2-globin gene creates a PstI cutting site. PMID- 11939520 TI - Unstable Hb Perth in a Taiwanese subject: a T-->C substitution at codon 32 of the beta-globin gene creates an MspI site. PMID- 11939522 TI - Hb Ube-2 in a Taiwanese subject: an A-->G substitution at codon 68 of the alpha2 globin gene. PMID- 11939523 TI - Second International Conference on Trace Element Speciation in Biomedical, Nutritional and Environmental Sciences. PMID- 11939524 TI - Overview of the methodology of nuclear analytical techniques for speciation studies of trace elements in the biological and environmental sciences. AB - Recent achievements in speciation studies of trace elements in the biological and environmental sciences by nuclear analytical techniques, mainly molecular activation analysis, position-sensitive spectrometry with a variety of exciting sources, and synchronous radiation-based analytical techniques (although radioisotope or enriched stable isotope-based speciation techniques are also used), particularly in our laboratory, are outlined. In this paper the merits and drawbacks of the nuclear analytical techniques are discussed, as are reagent blanks, contamination, and artifacts. PMID- 11939525 TI - Speciation analysis of trace elements in the brains of individuals with Alzheimer's disease with special emphasis on metallothioneins. AB - A speciation analysis of protein-bound elements in the cytosol of human brain was achieved by size exclusion chromatographical separation of the biomolecules and on-line detection of the metal profiles in the eluate by hyphenated inductively coupled plasma-mass spectrometry. Post-mortem samples from Alzheimer's disease brains and from brains of a control group were investigated to elucidate changes in the trace element distribution during the pathological process. Special attention was paid to the metallothioneins (MT) - cysteine-rich, metal-binding proteins of low molecular weight, existing in several isoforms. The isoform MT-3 is found especially in the brain and has a growth inhibition function on neurons. The MT peaks were identified in the element profiles. For this purpose, the metal binding capability and the heat stability of MT were taken into consideration. For verification, a comparison with pure MT-3 was carried out and further biochemical and analytical methods were applied to the fractions of the chromatographical run. A comparison between Alzheimer's disease and control brains showed a significant difference concerning the MT-1/-2 and MT-3 metal levels, leading to the assumption that there were oxidative processes having taken place in the Alzheimer's brain samples. PMID- 11939526 TI - Chemical speciation of plutonium in seawater. AB - Concentrations of particulate and dissolved plutonium in seawater were determined together with strong organic ligands (SOL) in particulate matter (PM). The concentration of particulate 239,240Pu in surface waters of the Western North Pacific ranged from 0.03 to 0.3 mBq m(-3) during the period from 1987 to 1997. The percentage of particulate Pu to total Pu in seawater was less than 10% in surface waters, whereas its portions were less than 1% in deep waters. In order to characterize particulate Pu in seawater, the relationships between particulate Pu, dissolved Pu and SOL concentrations in surface PM were examined. The result reveals that particulate Pu was linearly related to the SOL concentration independent of dissolved Pu concentration. Mass balance analysis suggests that a dominant chemical form of Pu in surface PM, which may exist as Pu(IV), is a complex with strong organic ligands in PM. PMID- 11939527 TI - Interaction of chemical species with biological regulation of the metabolism of essential trace elements. AB - Variations in the chemical speciation of dietary trace elements can result in the provision of different amounts of these micronutrients to the organism and might thus induce interactions with trace-element metabolism. The chemical species of Zn, Fe, Cu, and Mn can interact with other components of the diet even before reaching the site of absorption, e.g. by formation of poorly soluble complexes with phytic acid. This might considerably modify the amount of metabolically available trace elements; differences between absorptive capacity per se toward dietary species seems to be less important. Homeostasis usually limits the quantities of Zn, Fe, Cu, and Mn transported from the gut into the organism, and differences between dietary species are largely eliminated at this step. There is no homeostatic control of absorption of Se and I, and organisms seem to be passively exposed to influx of these micronutrients irrespective of dietary speciation. Inside the organism the trace elements are usually converted into a metabolically recognizable form, channeled into their biological functions, or submitted to homeostatically controlled excretion. Some dietary species can, however, be absorbed as intact compounds. As long as the respective quantities of trace elements are not released from their carriers, they are not recognized properly by trace element metabolism and might induce tissue accumulation, irrespective of homeostatic control. PMID- 11939529 TI - Trace cobalt speciation in bacteria and at enzymic active sites using emission Mossbauer spectroscopy. AB - 57Co emission Mossbauer spectroscopy (EMS) allows the chemical state of cobalt, as influenced by its coordination environment, to be monitored in biological samples at its physiological (trace) concentrations. To draw attention to EMS as a valuable tool for speciation of cobalt in biocomplexes, the process of cobalt(II) metabolism in cells of the plant growth-promoting rhizobacterium Azospirillum brasilense Sp245 was investigated using EMS of 57CoII-doped bacterial cells. EMS measurements also showed 57CoII-activated glutamine synthetase (GS, a key enzyme of nitrogen metabolism, isolated from this bacterium) to have two different cobalt(II) forms at its active sites, in agreement with data available on other bacterial GSs. Chemical after-effects following electron capture by the nucleus of the parent 57CoII during the 57Co- >57Fe transition, which contribute to the formation of a stabilised daughter 57FeIII component along with the nucleogenic 57FeII forms, are also briefly considered. PMID- 11939528 TI - Speciation and subcellular location of se-containing proteins in human liver studied by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and hydride generation-atomic fluorescence spectrometric detection. AB - Speciation of Se-containing proteins in the subcellular fractions of human liver was studied by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS PAGE) followed by hydride generation-atomic fluorescence spectrometric (HG-AFS) detection. It was found that about 24 kinds of Se-containing proteins existed in subcellular fractions of normal human liver. The molecular weights (MW) of the subunits were mostly in the range 20-30 kDa and 50-80 kDa. Major Se-containing protein fractions at 61 kDa and 21 kDa are probably selenoprotein P and glutathione peroxidase, respectively. The 54 kDa protein is probably a thioredoxin reductase, which is presented in nuclei, mitochondria, lysosome, microsome and cytosol. We noticed that the Se-containing protein with the lowest MW of 9.3 kDa only existed in lysosome. Most of the proteins have not been identified and would require further investigation to characterize them. The specific subcellular distributions of different Se-containing proteins suggest that they could play important biological roles in each organelle. PMID- 11939530 TI - Trace metal and mineral speciation of remediated wastes using electron microscopy. AB - Electron microscopic techniques, including scanning electron microscopy (SEM), transmission electron microscopy (TEM), and electron probe microanalyses (EPMA), were used to evaluate metal species and mineralogical phases associated with metal-bearing contaminated soil and industrial wastes that have been solidified and stabilized with Portland cement. Metals present in the wastes included arsenic, barium, cadmium, chromium, copper, lead, nickel, and zinc. In addition, mineral alterations and weathering features that affect the durability and containment of metals in aged remediated wastes were analyzed microscopically. Physical and chemical alteration processes identified included: freeze-thaw cracking; cracking caused by the formation of expansive minerals, such as ettringite and thaumasite; carbonation; and the movement of metals from waste aggregates into the surrounding cement matrix. Preliminary results show that although the extent of degradation after 6 years is considered slight to moderate, evaluations of durability and permanence of metals containment cannot be based on leaching and bulk chemistry analyses alone. The use of electron microscopic analyses is vital in studies that evaluate trace metal and mineral species and that attempt to predict the long-term performance of metal containment in solidified and stabilized wastes. PMID- 11939531 TI - Effect of different extraction procedures on the yield and pattern of Se-species in bacterial samples. AB - Investigations are described to extract Se-species from a bacterial sample. The five extraction methods investigated were: hot water, protease, lysozyme, lysozyme-protease, and HCl hydrolysis. The extraction efficiency was determined by comparing the total amounts of selenium in the sample after pressure digestion with the amounts extracted by the different methods described. Efficiencies were found to be only 1% (hot water), ca. 8% (protease, HCl hydrolysis) or ca. 12% (lysozyme, lysozyme-protease). The Se-peak patterns were compared after investigating the extracts with strong anion exchange chromatography-inductively coupled plasma mass spectrometry (SAX-ICP-MS). Most promising were the lysozyme assisted procedures, which showed the highest diversity of species. Here, in the protease-lysozyme approach, the protease seemed to break down species that had been extracted by lysozyme from the bacterial wall (murein sacculus). The other approaches seemed not to extract many species. Hot water extraction was completely unsuitable, extracting only low amounts of a single, unknown species. PMID- 11939532 TI - Speciation of heavy metals in environmental water by ion chromatography coupled to ICP-MS. AB - Biogenic (e.g. phytochelatins, porphyrins, DOM) as well as anthropogenic (e.g. NTA, EDTA, phosphonates) chelators affect the mobility and cycling of heavy metals in environmental waters. Since such chelators can form strongly bound anionic heavy metal complexes that are stable and highly mobile, anion-exchange chromatography coupled to ICP-MS was investigated. A narrow bore HPLC system was connected to a micro concentric nebuliser for in-line sample introduction. A new chromatographic procedure based on a synthetic hydrophilic quaternary ammonium anion exchanger in combination with nitrate as a strong eluent anion, and gradient elution, provided high separation selectivity and a large analytical window. Low detection limits (nmol L(-1)) were achieved by on-column matrix removal and sample preconcentration. This allowed the method to be successfully applied to different environmental research areas. In ecotoxicological studies of heavy metal effects on algae low concentrations of metal EDTA complexes were determined in nutrient solutions without interference from high (buffer) salt concentrations. In groundwater, infiltrated by a polluted river, mobile metal EDTA species were observed. In river water of different pollution levels beside CuEDTA other anionic Cu-complexes were found in nmol L(-1) concentrations. PMID- 11939533 TI - Investigation of the response of wood-rotting fungi to copper stress by size exclusion chromatography and capillary zone electrophoresis with ICP MS detection. AB - A method based on the coupling of size-exclusion chromatography (SEC) with inductively coupled plasma mass spectrometry (ICP MS) was developed for screening the changes in the bioligand composition of wood-rotting fungi as a function of their exposure to copper stress. Strains of four different species of wood rotting fungi: Phanerochaete chrysosporium, Schizophyllum commune, Daedalea quercina and Pleurotus ostreatus were examined. Only one, namely Ph. chrysosporium, showed any significant difference in terms of the fingerprint of Cu-binding ligands between control and exposed cells which suggest trapping of Cu(II) by cell walls as the only resistance mechanisms. In the case of Ph. chrysosporium the bioinduction of a new Cu-binding ligand was demonstrated. The presence of a new compound in the SE chromatographic fraction of interest was confirmed by CZE-ICP MS. Attempts to identify the new compound by electrospray MS/MS failed because of insufficient sensitivity. PMID- 11939534 TI - Multidimensional liquid chromatography with parallel ICP MS and electrospray MS/MS detection as a tool for the characterization of arsenic species in algae. AB - An analytical strategy was developed for the characterization of arsenic species in a Laminaria algae. The approach was based on multidimensional liquid chromatography (LC) including sample extract cleanup by size-exclusion LC, separation of arsenic species by anion-exchange LC, verification of the chromatographic purity of arsenic-containing fractions, and their further purification, if necessary, by reversed-phase (RP) HPLC. The complementarity of ICP MS, used as the chromatographic detector, and ES MS/MS, employed for the identification of the peaks observed, was demonstrated. The species found were: arsenosugar A 11.7+/-0.5 microg g(-1), AsV 10.9+/-2.1 microg g(-1), arsenosugar B 2.22+/-0.07 microg g(-1), arsenosugar D 1.5+/-1.2 microg g(-1), a newly detected arsenosugar 1.13+/-0.07 microg g(-1), arsenosugar C 0.61+/-0.04 microg g(-1), DMA 0.42+/-0.02 microg g(-1) and these accounted for >99% of the arsenic present. The identities of all the species, except the newly detected compound, were doubly checked by matching the retention times of chromatographically pure (after the 3rd LC dimension) species with standards and by ES MS/MS. PMID- 11939535 TI - Development and validation of capillary electrophoresis for the determination of selected metal ions in airborne particulate matter after sequential extraction. AB - A capillary electrophoresis (CE) method has been developed and validated for the determination of the distribution of metals in particulate matter after three stage sequential extraction. Fe(II), Zn(II), Cu(II), Mn(II), and Cd(II) ions were separated using a background electrolyte consisting of 200 mmol L(-1) ammonium acetate (pH 5.5), 0.5 mmol L(-1) 1,10-phenanthroline, 10 mmol L(-1) hydroxylamine hydrochloride, and 20% acetone. The method was validated with respect to specificity, linearity, detection limits, precision, and accuracy. Detection limits are at sub-microg L(-1) levels (tens of microg g(-1) of particulate matter) using pressure injection. The analytical procedure was checked by analyzing a standard reference material, NIST SRM 1648 Urban Particulate Matter. PMID- 11939536 TI - Improving selenium extraction by sequential enzymatic processes for Se-speciation of selenium-enriched Agaricus bisporus. AB - Sample preparation methods based on the use of proteolytic and cell wall digesting enzymes for the speciation analysis of selenized mushroom were investigated. The sample (Agaricus bisporus; 160 microg total Se per g sample) was grown on compost supplemented with selenized yeast. Experiments were carried out to elucidate the possible role of the cell wall digesting enzymes--Lysing enzyme and Driselase--in the improvement of extraction efficiency with and without inhibiting proteolysis during cell wall digestion. A 3-step procedure applying Lysing enzyme and pronase gave the highest extraction efficiency (89%); however, the best species recovery was achieved by a one-step proteolytic procedure. All the procedures of selenium speciation were controlled by independent ICP-AES analysis measuring the total amount of selenium. PMID- 11939537 TI - Iodine species and the endocrine system: thyroid hormone levels in adult Danio rerio and developing Xenopus laevis. AB - Recently a new approach for the analysis of iodinated organic species in human serum has been developed using liquid chromatography-inductively coupled plasma mass spectrometry (LC-ICP-MS). This method enables quantification of iodide, T4 and T3, as well as reverse T3 (rT3) and the synthetic precursors of TH, monoiodotyrosine (MIT), and diiodotyrosine (DIT) in a single injection. In this work, the LC-ICP-MS approach was used to analyze whole-body homogenates of adult male and female zebrafish (Danio rerio) and tadpoles of the African clawed frog (Xenopus laevis) at two different developmental stages (NF58 and 61) according to Nieuwkoop and Faber. The data demonstrate that the LC-ICP-MS method was successful at measuring I-, MIT, DIT, T4, T3, and rT3 in these two species. Furthermore, the method also detected five additional iodinated compounds which are currently unidentified. PMID- 11939538 TI - Selenium speciation by HPLC with tandem mass spectrometric detection. AB - An HPLC/MS-MS method was developed for the analysis of selenium species. Tandem mass spectrometry (MS-MS) was chosen as a detector to provide structural and molecular information allowing the identification of species, which are not commercially available as standards. A new separation method for selenium species was developed, based on porous graphitic carbon (PGC) as the stationary phase. The applicability of the optimized HPLC/MS-MS system was demonstrated by the analysis of a mixture containing Se-methyl-selenocysteine, selenomethionine, selenocystine, selenoethionine and selenocystamine. All peaks were baseline resolved and eluted within 16 min. Positive ionization led to higher intensities than negative ionization. Signal suppression tests showed that electrospray ionization (ESI) is a more effective ionization method than atmospheric pressure chemical ionization (APCI) for selenium species in a matrix containing pentafluoropropionic acid, heptafluorobutyric acid or ammonium formate. Comparative experiments with a triple quadrupole mass spectrometer (Quattro LC) and a time-of-flight instrument (Q-Tof-2) showed a 20 fold higher mass resolution of the latter mass spectrometer (m/Am= 5000) and significantly lower intensities for analyte signals as well as background noise compared to the triple quadrupole instrument. MS-MS spectra of the investigated selenium species including characteristic fragmentation patterns are presented. PMID- 11939539 TI - Sampling and processing of biopsy samples for speciation studies of cytosolic metalloproteins. AB - Studies to specify various metalloproteins in cell cytosol of tissues using chromatographic separation methods and plasma mass spectrometry for element detection require a careful step by step sample preparation. These steps involve (i) bioptic sample removal from tissue, (ii) extraction and preparation of the tissue supernatant, (iii) the chromatographic separation of the proteins, and finally (iv) the on-line transfer of the column eluate into the plasma mass spectrometer. Each of the analytical steps has to be carefully monitored to avoid undesired changes in the sample composition which could be caused by enzymatic and/or oxidative processes as well as by external element contamination. This presentation introduces a concept to ensure that environmental element contamination does not occur to bias the analytical results. PMID- 11939540 TI - Monitoring of total antimony and its species by ICP-MS and on-line ion chromatography in biological samples from patients treated for leishmaniasis. AB - Results from a study are reported in which patients with leishmaniasis were monitored by whole blood, blood plasma, urine, and hair analysis, before, during, and after intramuscular administration of N-methyl meglumine antimoniate. Quadrupole ICP-MS was used for the detection of antimony and on-line ion chromatography for the separation of its species. After typically 30 consecutive daily injections of 5 mg antimony per kg of body weight, Sb concentrations of up to 250 microg L(-1) in whole blood and plasma, and 60 mg of Sb per gram of creatinine in urine, were measured 24 h after drug administration. Antimony in hair samples of these patients showed concentrations of up to 24 microg g(-1). Speciation studies of Sb5+ and Sb3+ in drug, urine, and plasma samples were performed by ion chromatography using a Hamilton PRP-100X anion exchange column and EDTA (2 or 20 mM, pH 4.7) as the mobile phases. Repeatability of elution time and peak area measurements for a 0.125 ng spike were <1.2% and <3.5%, respectively. Method detection limits for both species, using a 1:10 diluted urine or plasma sample, were typically 1.6 microg L(-1). The procedure was capable of separating the very intense drug peak from its inorganic species, thus permitting the first studies on the bio-transformation of N-methyl meglumine antimoniate to Sb5+ and Sb3+ in the human body. PMID- 11939541 TI - Sulfur speciation in hard coal by means of a thermal decomposition method. AB - A new method for the determination of organic and pyritic sulfur in hard coal is presented. The method is based on controlled thermal decomposition of coal sample in oxygen-free and oxygen atmospheres. The results for sulfur liberated in an argon atmosphere at temperatures up to 773 K were close to organic sulfur contents (Sorg), although owing to the definition of 'organic sulfur' the values were not directly comparable. Sorg contents are calculated from the difference between total sulfur content in coal and contents of this element in the form of sulfides, sulfates and pyrites. Sulfur contents, found in the second stage of analysis, were close to pyritic sulfur contents. The difference between total sulfur content and the sum of sulfur values obtained in stages I and II corresponded to sulfur contents in those samples which were neither decomposed nor oxidized at temperatures up to 1173 K. Although not comparable with such conventional concepts for industrial purposes these data are attractive due to the ease and rapidity of the new method for the control of sulfur streams in industrial processes. PMID- 11939542 TI - A new macrocyclic polystyrene-based sensor for chromium (III) ions. AB - A new tetradentate dihydrogen perchlorate macrocyclic ligand (2,4,9,11 tetraphenyl-1,5,8,12-tetraazacyclotetradeca-1,4,8,11-tetraene dihydrogen perchlorate) was prepared and characterised. The macrocycle behaves as a selective chelating ion-exchanger for some metal ions. The polystyrene-based membrane electrode is found to exhibit quite promising selectivity for Cr3+ ions. It can be used to estimate chromium concentrations in the range 3.16x 10(-6) 1.00x10(-1) M with a near-Nernstian slope of 17.5 mV per decade of concentration between pH 3.0 to 6.5. The electrode is found to possess a fast response time of 15 s and was used over a period of three months with good reproducibility (s = +/ 0.3 mV). The selectivity coefficient values for mono-, di- and trivalent cations indicate excellent selectivity for Cr3+ ions over a large number of other cations. Anions such as Cl- and SO4(2-) do not interfere and the electrode also works satisfactorily in a mixed organic-water solution. The sensor has been used as an indicator electrode for the potentiometric titration of Cr3+ with EDTA. The practical utility of the membrane sensor has also been demonstrated in solutions contaminated with detergents (CTAB and SDS). Above all, the membrane sensor has been very successfully used to determine Cr3+ in some foods. PMID- 11939543 TI - On-line post-capillary affinity detection of immunoglobulin G for capillary zone electrophoresis. AB - To address the quality issues of antibody manufacturing, post-capillary affinity detection of immunoglobulin G (IgG) is developed for capillary zone electrophoresis. In analogy to a two-dimensional separation system, capillary zone electrophoresis (CZE), as the first dimension, resolves IgG variants based on their differences in molecular structure. IgG variants separated by CZE are discriminated against other serum and cellular proteins by affinity complex formation with protein A binding fragment in a post-capillary reactor. The analytical power of post-capillary affinity detection is demonstrated for rapid and selective heterogeneity analysis of human IgG subclasses and monoclonal antibodies in complex sample matrices. By comparing with pre-capillary formation of affinity complexes between IgG and protein A, post-capillary affinity detection clearly exhibit greater resolving power for examining IgG microheterogeneity. Affinity complex formation prior to CZE analysis, however, has the advantage of lower detection limits. Detection limits suffer with post capillary affinity detection because of the high fluorescence background contributed by the fluorescently labeled protein A in the post-capillary reactor, and the need to determine a small change in the background level upon complex formation. PMID- 11939544 TI - Estimation of drug-protein binding parameters on assuming the validity of thermodynamic equilibrium. AB - This contribution focuses the reader's attention on the pitfalls usually emerging during the phase of evaluation of experimental data of drug-protein binding studies. To overcome the occurrence of problem(s) apparently defying solution, the concept of "affinity spectra" is recommended to be implemented for data evaluation. A (general) "binding study protocol" is also suggested, which can prevent the formation of inadequate conclusions and the generation of unrealistic drug-protein binding parameters. PMID- 11939545 TI - Solute complexation degree with human serum albumin: biochromatographic approach. AB - A mathematical model was developed for the study of the D,L-dansylamino acid retention mechanism in reversed-phase liquid chromatography using a C18 column as a stationary phase and human serum albumin (HSA) as an eluent modifier. The solute retention factor is dependent on the HSA concentration in the eluent as well as the binding constant of the guest-HSA complex. A determination of the degree of complexation n(c) (the percent of the complexed guest) could be carried out. Different Van 't Hoff plot shapes of the degree of complexation were observed with different eluent pH, confirming a change in the solute complexation mechanism for physiological pH (between 7-7.5). Enthalpy-entropy compensation was also analysed in relation to this mathematical model to confirm the solute complexation behavior with HSA. These results finally confirmed that at physiological pH and temperature (approximately 35 degrees C) values the HSA was in a favorable structural conformation for its binding with a great majority of drugs. PMID- 11939546 TI - Role of the magnesium cation on antihypertensive molecule-human serum albumin binding: affinity chromatography approach. AB - The role of the Mg2+ cation on antihypertensive molecule binding on human serum albumin (HSA) was studied by affinity chromatography. The thermodynamic data corresponding to this binding were determined for a wide range of Mg2+ concentrations (c). For the nifedipine molecule, an increase in the Mg2+ concentration produced a decrease in binding due to a decrease in the electrostatic interactions. For verapamil and diltiazem, which have the highest solvent accessible surface area, the solute binding on HSA was divided into two Mg2+ concentration regions. For a low c value below c(c) (approximately 1.6 mmol/l), the binding dependence with c was similar to that of nifedipine. For c above c(c) the hydrophobic effect created in the bulk solvent associated with a decrease in the van der Waals interactions between the solute molecule and the HSA implied a decrease in its binding. These results showed that for patients with hypertension, an Mg2+ supplementation during treatment with these antihypertensive molecules can increase the active pharmacological molecule concentration. PMID- 11939547 TI - Stereoselective binding of 2-(4-biphenylyl)-3-substituted-3-hydroxypropionic acids on an immobilised human serum albumin chiral stationary phase. AB - A series of 2-(4-biphenylyl)-3,3'-hydroxy-substituted phenyl propionic acid, with anti-inflammatory properties, bearing two chiral centres, were studied by HPLC upon HSA-CSP (human serum albumin-based chiral stationary phase). The compounds were analysed in their stereoisomeric erythro and threo forms. The study involved the enantioselective analysis on HSA-CSP, the determination of the racemate lipophilicity (log k'(w)), a QSRR (quantitative structure-retention relationship) analysis and CD study for the assessment of the absolute configuration of the most retained enantiomer. Lipophilicity was found to be an important factor affecting the affinity of the compounds for the HSA stationary phase, but electronic properties seemed to play a role. The position of the substituent of the phenyl group on carbon 3 was found important to modulate stereoselective interaction, the highest value of enantioselectivities being found for the erythro ortho-substituted phenyl derivatives. The previously proposed two steps mechanism of enantiodiscrimination for cyclohexylphenyl substituted derivatives was confirmed for this series of derivatives bearing the biphenylyl moiety. PMID- 11939548 TI - Use of an immobilised human serum albumin HPLC column as a probe of drug-protein interactions: the reversible binding of valproate. AB - The reversible binding of valproate to human serum albumin determines a decrease of the binding of ligands that selectively bind to site I, site II, and bilirubin binding site. The binding inhibition was followed by displacement chromatography methodology using increasing concentrations of the competitor, i.e. valproate, in the mobile phase. Significant binding inhibition was observed for drugs binding at site I and site II. The greater displacement was observed for the more retained enantiomer of benzodiazepines and profens. A reduction of the affinity was observed also in the case of phenol red, this compound being selected as representative of bilirubin binding site. Difference circular dichroism spectroscopy was also used to characterise the binding of valproate to human serum albumin. This antiepilectic drug was proved to affect the binding at site I, II, and bilirubin binding site. The data have physiological relevance because significant inhibition of the binding resulted at clinic concentrations of valproate. PMID- 11939549 TI - Binding of human serum albumin to silica particles by means of polymers: a liquid chromatographic study of the selectivity of resulting chiral stationary phases. AB - Chiral stationary phases obtained by immobilization of human serum albumin (HSA) on various polymer-coated silicas were tested to resolve DL-tryptophan, DL-NBP, RS-oxazepam and RS-warfarin racemic mixtures. HSA immobilized on anion exchangers [quaternized poly(vinylimidazole)-coated silica] was highly selective. Stable and selective chiral stationary phases were also prepared by covalent binding of HSA to silica particles via reactive-polymers. Poly(acryloyl chloride), poly(methacryloyl chloride) and poly(vinyl chloroformate) derivatives were compared. Parameters that govern the selectivity of resulting chiral supports were evaluated, especially the orientation of HSA after immobilization, the mobility of polymer chains and the number of covalent linkages between the protein and the polymer. PMID- 11939550 TI - Use of the hummel and dreyer method for the study of nucleotide binding on chloroplast ATPase CF1. AB - The study of the binding of the nucleotides ADP and ATP on the exchangeable sites of chloroplast ATPase CF1 has been carried out by the Hummel and Dreyer method applied to HPLC. It has been shown that this method was well fitted to the problem: rapidity of exchange, absence of noticeable modification after binding, presence of a constant concentration of ligand during the chromatography, which stabilizes these low affinity complexes. The dissociation constants of binding of ADP, ATP and of their magnesium salt complexes have been determined. In order to measure the simultaneous binding of ADP and ATP when present in mixture, we have modified the method by using an anion-exchange column in place of the gel filtration column: the two nucleotides were easily separated, while the binding on the protein was unchanged. The extension of this method to the reversed-phase chromatography could also be considered for the binding of hydrophobic ligands. PMID- 11939551 TI - Plasma protein binding study of oxybutynin by high-performance frontal analysis. AB - Plasma protein binding of oxybutynin (OXY) was investigated quantitatively and enantioselectively using high-performance frontal analysis (HPFA). An on-line HPLC system which consists of HPFA column, extraction column and analytical column was developed to determine the unbound concentrations of OXY enantiomers in human plasma, in human serum albumin (HSA) solutions, and in human alpha1-acid glycoprotein (AGP) solutions. OXY is bound in human plasma strongly and enantioselectively. The bound drug fraction in human plasma containing 2-10 microM (R)- or (S)-OXY was higher than 99%, and the unbound fraction of (R)-OXY was 1.56 times higher than that of (S)-isomer. AGP plays the dominant role in this strong and enantioselective plasma protein binding. The total binding affinities (nK) of (R)- and (S)-OXY to AGP were 6.86 x 10(6) and 1.53 x 10(7) M( 1), respectively, while the nK values of (R)- and (S)-OXY to HSA were 2.64 x 10(4) and 2.19 x 10(-4) M(-1), respectively. The binding affinity of OXY to AGP is much higher than that to HSA, and shows high enantioselectivity (SIR ratio of nK values is 2.2). It was found that both enantiomers are bound competitively at the same binding site on an AGP molecule. The binding property between OXY and low density lipoprotein (LDL) was investigated by using the frontal analysis method incorporated in high-performance capillary electrophoresis (HPCE/FA). It was found the binding is non-saturable and non-enantioselective. PMID- 11939552 TI - Binding study of desethyloxybutynin using high-performance frontal analysis method. AB - Plasma protein binding of N-desethyloxybytynin (DEOXY), a major active metabolite of oxybutynin (OXY), was investigated quantitatively and enantioselectively using high-performance frontal analysis (HPFA). An on-line HPLC system which consists of HPFA column, extraction column and analytical column was developed to determine the unbound concentrations of DEOXY enantiomers in human plasma, in human serum albumin (HSA) solutions, and in human alpha1-acid glycoprotein (AGP) solutions. DEOXY is bound in human plasma strongly and enantioselectively. The unbound drug fraction in human plasma samples containing 5 microM (R)- or (S) DEOXY was 1.19 +/- 0.001 and 2.33 +/- 0.044%, respectively. AGP plays the dominant role in this strong and enantioselective plasma protein binding of DEOXY. The total binding affinity (nK) of (R)-DEOXY and (S)-DEOXY to AGP was 2.97 x 10(7) and 1.31 x 10(7) M(-1), respectively, while the nK values of (R)-DEOXY and (S)-DEOXY to HSA were 7.77 x 10(3) and 8.44 x 10(3) M(-1), respectively. While the nK value of (S)-DEOXY is weaker than that of (S)-OXY (1.53 x 10(7) M( 1)), the nK value of (R)-DEOXY is 4.33 times stronger than that of (R)-OXY (6.86 x I0(6) M(-1)). This suggests that the elimination of an ethyl group weakens the binding affinity of the (S)-isomer because of the decrease in hydrophobicity, while the binding affinity of the (R)-isomer is enhanced by the decrease in steric hindrance. The total binding affinity of DEOXY to HSA is much lower than that of DEOXY-AGP binding as well as OXY-HSA binding (2.64 x 10(4) and 2.19 x 10(4) M(-1) for (R)-OXY and (S)-OXY, respectively). The study on competitive binding between OXY and DEOXY indicated that DEOXY enantiomers and OXY enantiomers are all bound competitively at the same binding site of AGP molecule. PMID- 11939553 TI - Determination of the affinity constants of recombinant human galectin-1 and -3 for simple saccharides by capillary affinophoresis. AB - The affinity constants of recombinant human galectin-1 and galectin-3 for sugars were determined by capillary affinophoresis. The monoliganded affinophore contains p-aminophenyl-beta-lactoside as an affinity ligand in the matrix of succinylglutathione and has three negative charges. An analysis of the mobility change of the lectins caused by the affinophore and its inhibition by neutral sugars allowed, for the first time, a determination of the affinity constants between the binding sites of the lectins and sugars. The relative magnitude of the affinity constants for each of the sugars in terms of dissociation constants found to be consistent with previously reported data on the concentrations of sugars that caused a 50% inhibition (I50) in the binding assay of the lectin to oligosaccharide-immobilized agarose beads but the absolute values of the dissociation constants were considerably smaller than the I50 values. Capillary affinophoresis indicated microheterogeneity of the lectin preparations and enabled the separate analysis of the affinity of each component simultaneously showing the advantage in using a separation method for analysis of bioaffinity. PMID- 11939554 TI - Study on the multiple sites binding of human serum albumin and porphyrin by affinity capillary electrophoresis. AB - Equations to describe the two sites binding between proteins and ligands were deduced. According to these equations, not only the binding constants, but also the mole fraction of proteins in different forms could be obtained. Using the published data on the interaction between human serum albumin (HSA) and three kinds of porphyrin (coproporphyrin (CP), uroporphyrin I (UP) and protoporphyrin (PP)), a further study on their binding was carried out. It was concluded that there may exist two binding sites with the binding constants at the first site, proved to be the preferential one, being 6.50 x l0(5), 1.94 x 10(6) and 8.94 x 10(5), respectively. In addition, it was also demonstrated that the two binding sites of HSA with CP and UP might be of different kinds, though those of HSA and PP were of the same kind but at different positions. PMID- 11939555 TI - High-performance affinity chromatography: a powerful tool for studying serum protein binding. AB - High-performance affinity chromatography (HPAC) is a method in which a biologically-related ligand is used as a stationary phase in an HPLC system. This approach is a powerful means for selectively isolating or quantitating agents in complex samples, but it can also be employed to study the interactions of biological systems. In recent years there have been numerous reports in which HPAC has been used to examine the interactions of drugs, hormones and other substances with serum proteins. This review discusses how HPAC has been used in such work. Particular attention is given to the techniques of zonal elution and frontal analysis. Various applications are provided for these techniques, along with a list of factors that need to be considered in their optimization and use. New approaches based on band-broadening studies and rapid immunoextraction are also discussed. PMID- 11939556 TI - Immobilized-biomembrane affinity chromatography for binding studies of membrane proteins. AB - Analyses of specific interactions between solutes and a membrane protein can serve to characterize the protein. Frontal affinity chromatography of an interactant on a column containing the membrane protein immobilized in a lipid environment is a simple and robust approach for series of experiments with particular protein molecules. Regression analysis of the retention volumes at a series of interactant concentrations shows the affinity of the protein for the interactant and the amount of active binding sites. The higher the affinity, the fewer sites are required to give sufficient retention. Competition experiments provide the affinities of even weakly binding solutes and the non-specific retention of the primary interactant. Hummel and Dreyer size-exclusion chromatography allows complementary analyses of non-immobilized membrane materials. Analyses of the human facilitative glucose transporter GLUT1 by use of the inhibitor cytochalasin B (radioactively labeled) and the competitive substrate D-glucose (non-labeled) showed that GLUT1 interconverted between two states, exhibiting one or two cytochalasin B-binding sites per two GLUTI monomers, dependent on the membrane composition and environment. Similar analyses of a nucleoside transporter, a photosynthetic reaction center, nicotinic acetylcholine receptors and a P-glycoprotein, alternative techniques, and immobilized-liposome chromatographic approaches are presented briefly. PMID- 11939557 TI - Immobilized receptor- and transporter-based liquid chromatographic phases for on line pharmacological and biochemical studies: a mini-review. AB - This review addresses the synthesis and characterization of two different types of receptor-based liquid chromatographic supports, one based upon a trans membrane ligand gated ion channel receptor (the nicotinic acetylcholine receptor) and the other a soluble nuclear receptor (the estrogen receptor). In addition, studies with the P-glycoprotein transporter are also reported. The nicotinic receptor was immobilized via hydrophobic insertion into the interstitial spaces of an immobilized artificial membrane (IAM) stationary phase. the estrogen receptor was tethered to a hydrophilic stationary phase and the membranes containing the Pgp transporter were coated on the surface of the IAM stationary phase. The stationary phases were characterized using known ligands and substrates for the respective non-immobilized proteins. The results from zonal and frontal chromatographic experiments demonstrated that the stationary phases could be used to determine binding affinities (expressed as dissociation constants, Kd,'s) and to resolve mixtures of ligands according to their relative affinities. In addition. competitive ligand binding studies on the P-glycoprotein based stationary phase have established that this phase can be used to identify and characterize competitive displacement and allosteric interactions. These studies demonstrate that immobilized-receptor phases can be used for on-line pharmacological studies and as rapid screens for the isolation and identification of lead drug candidates from complex biological or chemical mixtures. PMID- 11939558 TI - Quantitative structure-retention relationships in affinity high-performance liquid chromatography. AB - In this report the affinity high-performance liquid chromatography data, which were determined on silica-based human serum albumin, alpha1-acid glycoprotein, keratin, collagen, melanin, amylose tris(3,5-dimethylphenylcarbamate), and basic fatty acid binding protein columns, are discussed. Using a quantitative structure retention relationship (QSRR) approach the affinity data were interpreted in terms of structural requirements of specific binding sites on biomacromolecules. The unique chromatographic properties of immobilized artificial membrane and cholesterol stationary phases were also analyzed from the point of view of mimicking biological processes. It has been demonstrated that chemometric processing of appropriately designed sets of chromatographic data derived in systems comprising biomolecules provides information of relevance for molecular pharmacology and rational drug design. PMID- 11939559 TI - Applications of affinity chromatography to the study of drug-melanin binding interactions. AB - This short review reports on progresses in the study of drug-melanin interactions using the technique of affinity chromatography. Melanins are natural or synthetic pigments derived from the oxidation and polymerization of various precursors including L-dopa, tyrosine and cystein. Accumulation of toxic compounds, drugs, and metal ions in pigmented tissues through reversible binding to melanin has been linked to chronic toxicity. Affinity chromatography using chromatographic stationary phases based on physically adsorbed or chemically bonded melanin provides a useful tool for studying the interactions of small molecules and metal ions with melanin PMID- 11939560 TI - Antibody-antigen binding study using size-exclusion liquid chromatography. PMID- 11939561 TI - Estimation of binding constants by capillary electrophoresis. AB - Capillary electrophoresis (CE) has become a useful technique for measuring binding constants. This review is focused on recent trends in the estimation of binding constants by affinity CE. First, we introduce several mathematical equations in which it is assumed that the stoichiometry of the binding between drug and protein is 1:1 as a simple model. In order to calculate accurate binding constants by affinity CE, several experimental considerations are described in this review. In addition, some recent methodologies, such as partial filling technique and multiple-step ligand injection method, are introduced. Among research publications within 3 years, recent applications for determining binding constants are reviewed. PMID- 11939562 TI - Antigen-antibody interactions in capillary electrophoresis. AB - Immunoreactions in combination with separations by capillary electrophoresis (CE) are increasingly being used to quantitate specific analytes in biological fluids. Both competitive and non-competitive approaches have been used for the purpose and, in selected cases, now compare favorably with conventional quantitative immunoassays with respect to concentration limits of detection. CE is also a useful method to evaluate antigen-antibody binding on-line and offers unique possibilities for binding constant estimates, also for weakly binding antibodies and antibody fragments. In this review we cover recent developments in the use of antigen-antibody interactions in conjunction with CE and conclude that continued development of miniaturization, on-line preconcentration and more sensitive detection schemes will contribute to the further dissemination of CE-based immunoassays building on already established affinity CE approaches. PMID- 11939563 TI - Distribution of repetitive sequences on the leading and lagging strands of the Escherichia coli genome: comparative study of Long Direct Repeat (LDR) sequences. AB - In the present study, we developed a method for detecting sequences whose similarity to a target sequence is statistically significant and we examined the distribution of these sequences in the E. coli K-12 genome. Target sequences examined are as follows: (i) short repeat: Crossover hot-spot instigator (Chi) sequence, replication termination (Ter) sequence, and DnaA binding sequence (DnaA box); (ii) potential stem-loop structure repeats: palindromic unit (PU), boxC sequences, and intergenic repeat unit (IRU); (iii) potential RNA coding repeats: rRNAs, PAIR, TRIP, and QUAD; and (iv) potential protein coding repeats: insertion elements (ISs) and Long Direct Repeats (LDRs). We also examined the distribution of these sequences on leading and lagging strands. We obtained another four statistically significant LDR sequences with more than 187 bp matched to LDR-A near the LDR loci, suggesting that these regions might be used as high recombination hot spots for LDR. Adaptation of individual LDRs to E. coli genome is also discussed on the basis of codon usage. PMID- 11939564 TI - Positional cloning of rice semidwarfing gene, sd-1: rice "green revolution gene" encodes a mutant enzyme involved in gibberellin synthesis. AB - A rice semidwarfing gene, sd-1, known as the "green revolution gene," was isolated by positional cloning and revealed to encode gibberellin 20-oxidase, the key enzyme in the gibberellin biosynthesis pathway. Analysis of 3477 segregants using several PCR-based marker technologies, including cleaved amplified polymorphic sequence, derived-CAPS, and single nucleotide polymorphisms revealed 1 ORF in a 6-kb candidate interval. Normal-type rice cultivars have an identical sequence in this region, consisting of 3 exons (558, 318, and 291 bp) and 2 introns (105 and 1471 bp). Dee-Geo-Woo-Gen-type sd-1 mutants have a 383-bp deletion from the genome (278-bp deletion from the expressed sequence), from the middle of exon 1 to upstream of exon 2, including a 105-bp intron, resulting in a frame-shift that produces a termination codon after the deletion site. The radiation-induced sd-1 mutant Calrose 76 has a 1-bp substitution in exon 2, causing an amino acid substitution (Leu [CTC] to Phe [TTC]). Expression analysis suggests the existence of at least one more locus of gibberellin 20-oxidase which may prevent severe dwarfism from developing in sd-1 mutants. PMID- 11939565 TI - Conservation of translation initiation sites based on dinucleotide frequency and codon usage in Escherichia coli K-12 (W3110): non-random distribution of A/T-rich sequences immediately upstream of the translation initiation codon. AB - Dinucleotide frequencies are useful for characterizing consensus elements as a minimum unit of nucleotide sequence because the neighborhood relations of nucleotide sequences are reflected in dinucleotides. Using a consensus score based on dinucleotide frequencies and intra-species codon usage heterogeneity, denoted by the Z1 parameter, we report the relationship between nucleotide conservation at the translation initiation sites of genes in the Escherichia coli K-12 genome (W3110) and codon usage in its downstream genes. Significant positive correlations were obtained in three regions centered at -13, -4, and +7, which correspond to the Shine-Dalgarno element, the A + T element immediately upstream of the translation initiation site, and the downstream box, respectively. PMID- 11939566 TI - Binding of 5-bromouracil-containing S/MAR DNA to the nuclear matrix. AB - Substitution of thymine with 5-bromouracil in DNA is known to change interaction between DNA and proteins, thereby inducing various biological phenomena. We hypothesize that A/T-rich scaffold/nuclear matrix attachment region (S/MAR) sequences are involved in the effects of 5-bromodeoxyuridine. We examined an interaction between DNA containing an intronic S/MAR sequence of the immunoglobulin heavy chain gene and nuclear halos prepared from HeLa cells. Upon substitution with 5-bromouracil, the S/MAR DNA bound more tightly to the nuclear halos. The multi-functional nuclear matrix protein YY1 was also found to bind more strongly to 5-bromouracil-substituted DNA containing its recognition motif. These results are consistent with the above hypothesis. PMID- 11939567 TI - Kaposi sarcoma update: clinical-molecular correlations. PMID- 11939568 TI - Hyaluronan metabolism in rat tail skin following blockage of the lymphatic circulation. AB - This study was undertaken to explore the effects of lymphatic blockage on the metabolism of hyaluronan in the skin. In initial experiments, [3H] hyaluronan was injected subcutaneously into the tail skin of rats that either had no surgical intervention (control) or into those that had their lymphatic drainage blocked two hours earlier (acute lymphedema) or after the lymphatics had been blocked for three months (chronic lymphedema). The removal of tritiated hyaluronan from the injection sites was determined by the appearance of [3H] in the plasma. The results showed that the clearance of injected hyaluronan was delayed in rats with lymphatic blockage. The half- life of injected hyaluronan in the controls was approximately 70-75 hr, compared with approximately 105-110 hr in the lymph blocking rats. The levels of radioactivity in the plasma from rats with both acute and chronically blocked lymphatics were lower than that of control rats during the entire follow up period. In addition, biochemical analysis revealed that there was a significant increased amount of hyaluronan in the tail skin three months after lymphatic blocking. These results suggest that lymph absorption is an important factor in the transport of hyaluronan from the interstitium. Blockage of regional draining lymphatics likely impairs the catabolism of hyaluronan, which stagnates in skin tissue. PMID- 11939569 TI - Limb volume reduction after physical treatment by compression and/or massage in a rodent model of peripheral lymphedema. AB - Lack of a standardized experimental counterpart of peripheral lymphedema (LE) in a small animal has hampered research into treatment of this debilitating condition. We recently refined a rodent model consisting of radical unilateral lymphatic/nodal groin excision in conjunction with a circumferential integumental gap, followed by regional irradiation of the groin to reproduce stable unilateral hindlimb LE (1). In the current study, Wistar-Fuzzy rats with established right hindlimb LE, were subdivided into five groups and subjected to one of the following daily physical regimens over a 5-day period: pneumatic compression pumping at 30 torr (PCP); low-stretch multi-layered compressive bandaging using Coban (CB); manual lymphedema drainage (MLD) or a light massage consisting of stationary circular motions using the fingertips; combined physiotherapy (CPT consisting of MLD + CB); and a no treatment or control group (CTRL). Hindlimb and LE volumes were serially measured before and after treatment. Whereas CTRL showed progressive worsening of hindlimb swelling, PCP, CB, CPT and MLD each produced similar and substantial edema reduction over the 5 day interval, PCP, CB and CPT induced vacillating edema reduction which, however, exceeded rebound swelling on a daily basis. MLD, on the other hand, showed a steady gradual daily decline in LE volume. PMID- 11939570 TI - Trends in the evaluation of lymphedema. AB - This study reviews the criteria and the various measurements available for evaluation of a patient with peripheral lymphedema based on a systematic literature search using Medline. Each clinician needs to be more aware of the measuring technique chosen to determine limb swelling with the inherent limitations of each if errors in calculations or misinterpretations are to be avoided. Determining the patient's symptoms (subjective) and limb changes (objective) are both importantfor evaluating response to treatment. PMID- 11939571 TI - Absence of HHV-8 infection in 37 patients with peripheral lymphedema. PMID- 11939572 TI - Traumatic injury of the thoracic duct. AB - Injuries to the thoracic duct are infrequent but may become life-threatening when chylous leakage persists. This report describes 6 patients with such injuries in whom the leakage resolved spontaneously in one, was corrected using microsurgical lymphatic repair or lymphatic-venous anastomosis in two, successfully treated either by ligation of the thoracic duct or insertion of a peritoneovenous shunt in two, and was eventually controlled after bilateral pleurodesis and thoracic duct ligation by insertion of a peritoneo-venous shunt in one. Conventional lymphography is superior to lymphoscintigraphy and is usually required to document disruption of the thoracic duct. PMID- 11939573 TI - Axillary dissection for breast cancer. PMID- 11939574 TI - Suppressed proliferative response of spleen T cells from metallothionein null mice. AB - To investigate the role of metal-binding protein, metallothionein (MT), in lymphocyte activation, the mitogen-induced proliferation of freshly isolated spleen cells was compared among MT-I, II null, and control 129/Sv mice. Spleen cells from MT null mice exhibited a markedly reduced proliferation compared with control cells when stimulated by concanavalin A or anti-CD3(epsilon) mAb, but not by lipopolysaccharide, indicating that only the response of T cells to mitogens was suppressed in MT null mice. Flow cytometric analysis of unstimulated spleen cells demonstrated no significant difference in the relative percentages of either B220+ and CD3+ cells or CD4+ and CD8+ cells between the two strains of mice. The production of interleukin (IL)-2 by MT null spleen cells after the stimulation by anti-CD3(epsilon) mAb was lower than that of control spleen cells, especially within 24 hr after the stimulation. The addition of IL-2 recovered the proliferation of MT null spleen cells to the control level. The reduced proliferative response to mitogenic stimulation of MT null T cells was confirmed by using purified splenic T cells. These results suggest that the MT expressed at basal level in the splenocytes plays an important role in T cell mitogen-induced proliferative response, probably by positively regulating the production of IL-2. PMID- 11939575 TI - Association of a regulatory gene, slyA with a mouse virulence of Salmonella serovar Choleraesuis. AB - The influence of slyA gene, originally found in Salmonella serovar Typhimurium as a regulatory gene for the expression of virulence genes, on a mouse virulence of S. serovar Choleraesuis was investigated by using an slyA-defective mutant. The defective mutant was constructed by the insertion of a kanamycin-resistance gene (aph) into the cloned slyA gene, and the homologous recombination with the intact slyA gene on the chromosome. The mutant strain showed the LD50 value for BALB/c mouse approximately 10(5) higher than that of the parent strain. The increase of the LD50 value was the same order as that shown by the mutation of the slyA gene of S. serovar Typhimurium, although LD50 of the wild-type strain of S. serovar Choleraesuis was 40-fold higher than that of S. serovar Typhimurium. The time course of infection observed in the mice organs also proved the clear difference of the virulence between the parent and the mutant strains. These results suggested that the slyA gene product functions as a virulence-associated regulator also in S. serovar Choleraesuis. PMID- 11939576 TI - Effect of anticoagulants in colorimetric assay for basic carboxypeptidases. AB - Carboxypeptidases (CP) in plasma and sera serve as regulators of anaphylatoxins such as C3a and C5a. The activity of CP can be measured by determining hippuric acid after cleavage of the small synthetic substrate hippuryl-L-arginine. Although a colorimetric assay is convenient for determining hippuric acid generated by CP, we noticed that some anticoagulants, such as citrate, interfere with the color development of the reagents used. EDTA and heparin provide an appropriate value. EGTA used as anticoagulant also provides an appropriate value. Therefore, concentration of citrate in samples should be controlled to be constant for background subtraction. PMID- 11939577 TI - Engineering of a Sagiyama alphavirus RNA-based transient expression vector. AB - Sagiyama virus (SAGV), a strain of Getah virus in the genus Alphavirus in the family Togaviridae, has a broad host range in vertebrates and invertebrates but is not pathogenic for humans. We engineered the SAGV genome as an efficient transient expression vector using the full-length infectious cDNA clone pSAG2 as the background. A green fluorescent protein (GFP) gene was used as a reporter gene and expressed from a subgenomic mRNA. When the GFP gene was placed downstream of the intact capsid protein gene or an internally deleted capsid protein gene encoding the N-terminal 9 amino acids and C-terminal 149 amino acids, autoproteolysis occurred efficiently at the boundary site to release GFP from the N-terminally-fused capsid-protease domain. GFP was also expressed efficiently without the 5'-terminal region of the capsid protein gene, suggesting that SAGV capsid protein gene does not have a translation enhancer sequence. To provide structural proteins for pseudovirion formation, a nonviable mutant construct, pSAG2.3L, which contains a Gly-to-Leu substitution at the - 2 position of the nsP3/4 cleavage site, was used as a helper. GFP was expressed up to 50 pg from 1 X 10(6) BHK21 cells after inoculation of pseudovirions. The C6/36 mosquito cell was also a suitable host for a large scale expression of GFP using pseudovirions. In addition to high-level transient expression, safeness of SAGV should give an advantage over other alphavirus expression vectors. PMID- 11939578 TI - Inactivation of C3a and C5a octapeptides by carboxypeptidase R and carboxypeptidase N. AB - Pro-carboxypeptidase R (proCPR), also known as thrombin-activatable fibrinolysis inhibitor (TAFI), precursor of carboxypeptidase U and plasma carboxypeptidase B is present in plasma and following activation by thrombin/thrombomodulin and/or plasmin can remove arginine from the carboxyterminal of C3a and C5a. We have shown that this enzyme can remove terminal arginine from the C5a octapeptide much more efficiently than the classical anaphylatoxin inactivator, carboxypeptidase N (CPN). Since we have previously demonstrated that proCPR is significantly upregulated in the inflammatory state, this enzyme would appear to significantly contribute to the inactivation of C5a, the most potent of the complement derived anaphylatoxins. PMID- 11939579 TI - Molecular ecology of toxigenic Vibrio cholerae. AB - Toxigenic Vibrio cholerae is the etiological agent of cholera, an acute dehydrating diarrhea that occurs in epidemic form in many developing countries. Although V. cholerae is a human pathogen, aquatic ecosystems are major habitats of Vibrio species, which includes both pathogenic and nonpathogenic strains that vary in their virulence gene content. V. cholerae belonging to the 01 and 0139 serogroups is commonly known to carry a set of virulence genes necessary for pathogenesis in humans. Recent studies have indicated that virulence genes or their homologues are also dispersed among environmental strains of V. cholerae belonging to diverse serogroups, which appear to constitute an environmental reservoir of virulence genes. Although the definitive roles of the virulence associated factors in the environment, and the environmental selection pressures for V. cholerae-carrying virulence genes or their homologues is not clear, the potential for origination of new epidemic strains from environmental progenitors seems real. It is likely that the aquatic environment harbors different virulence associated genes scattered among environmental vibrios, which possess a lower virulence potential than the epidemic strains. The ecosystem comprising the aquatic environment, V. cholerae, genetic elements mediating gene transfer, and the mammalian host appears to support the clustering of critical virulence genes in a proper combination leading to the origination of new V. cholerae strains with epidemic potential. PMID- 11939580 TI - Immunohistochemical distribution of epithelioid cell, myofibroblast, and transforming growth factor-beta1 in the granuloma caused by Mycobacterium avium intracellulare complex pulmonary infection. AB - The present study was designed to evaluate the distribution of epithelioid cells, myofibroblasts, and TGF-beta1 in the formation of granuloma caused by Mycobacterium avium intracellulare complex (MAC) lung infection. A retrospective study was performed for 9 cases of positive MAC culture in which lung resections were performed between January 1989 and August 1999. Resected lung specimens were evaluated histologically and immunohistochemically for CD68 (stain for monocytes and macrophages, and epithelioid cells) and alpha-smooth muscle actin as well as vimentin (stain for myofibroblasts), and TGF-beta1 was performed. When granuloma was initially formed, no myofibroblasts were found, but as caseous necrosis appeared, the thin epithelioid cell layer was detected and the outer myofibroblast layer gradually became thick. In the cavitary wall, the layer of epithelioid cells and multinucleated giant cells surrounded necrosis, and was associated with the outer layer of myofibroblasts. In addition, the anti-TGF beta1 antibody stained the cytoplasm of epithelioid cells and multinucleated giant cells, preceding the advent of myofibroblasts. In summary, our present study evaluated distributions of epithelioid cells, myofibroblasts, and TGF-beta along with the morphogenesis of granuloma, and clearly demonstrated the immunohistochemical difference between granuloma with caseous necrosis and granulomas without caseous necrosis. PMID- 11939581 TI - Antibacterial properties of antimicrobial-finished textile products. AB - The antibacterial properties of five kinds of antimicrobial-finished textile products (AFTPs) were examined against Staphylococcus aureus, including methicillin-resistant (MRSA) strains and Pseudomonas aeruginosa, under wet and dry conditions. Textile products containing Ag. Zn. ammonium Zeolite and chitosan were found to be effective against methicillin-sensitive S. aureus (MSSA) for up to 6 hr of incubation under wet and dry conditions, and effective against MRSA for up to 24 hr of incubation only under wet conditions. Under dry conditions, however, all AFTPs were ineffective against one MRSA strain. When organic matter was added to the incubation mixture, textile products containing Ag. Zn. ammonium Zeolite and chitosan still showed antibacterial activities, but not as strongly. The results of this study suggested the following: (1) There are differences in antibacterial properties among commercially available AFTPs; (2) Determining effectiveness requires several hours of incubation; (3) Water content as an environmental factor can affect effectiveness; and (4) Some bacterial species and strains are not affected by AFTPs. The antibacterial properties of AFTPs in the clinical setting may be of limited value. PMID- 11939582 TI - Development of a new medium useful for the recovery of dermatophytes from clinical specimens by minimizing the carryover effect of antifungal agents. AB - Two surface-active compounds, egg lecithin and polysorbate 80, usually used as the deactivators of various preservatives were tested whether they also counteract either or all of the three major topical antifungal drugs, bifonazole (BFZ), lanoconazole (LCZ) and terbinafine (TBF). Both egg lecithin and polysorbate 80, when added to culture media up to final concentrations of 1.0 and 0.7%, respectively, antagonized the anti-dermatophytic activity of the three drugs in a concentration-dependent manner. A greater extent of antagonistic action was exerted when the two deactivators combined at their maximal levels tested were added; MIC's of BFZ were increased more than 30-fold and those of LCZ and TBF more than 200-fold compared with the values obtained in the absence of the deactivators. Using the agar medium supplemented with the combined deactivators, culture studies were carried out with skin tissues specimens taken from guinea pigs whose feet were infected with dermatophytes and subsequently treated with 1% topical preparations of the three antifungal drugs. The experimental data from this animal study demonstrated that the combined deactivators-supplemented medium yielded increased numbers of fungi compared with the basal medium. It looks, therefore, likely that the fungal recovery on the former medium more correctly reflects to actual fungal burden in the infected lesions than the latter. All these results suggest that the combined deactivators supplemented medium is more useful for mycological evaluation of therapeutic efficacy of imidazole and allylamine drugs against dermatophytoses in both preclinical and clinical studies. PMID- 11939583 TI - Antifungal pharmacodynamic characteristics of amphotericin B against Trichosporon asahii, using time-kill methodology. AB - We determined the MIC of amphotericin B against 45 Trichosporon asahii isolates from various clinical and environmental sources, and used in vitro time-kill methods to characterize the relationship between amphotericin B concentrations and MIC for four representative T. asahii isolates. Amphotericin B had concentration-dependent antifungal activity. MICs ranged from 0.5 to 16 microg/ml, and most T. asahii isolates (76%, 34/45) were inhibited at safely achievable amphotericin B serum concentrations (< or = 2 microg/ml). However, 40% (18/45) of isolates were not killed at these concentrations (MFCs from 1.0 to 32 microg/ml). At concentrations > or = 2 x MIC, amphotericin B exhibited fungicidal activity (< 99.9% reduction in CFU) over a 12-hr time-period; the maximal effect was achieved at > or =4 x MIC. Susceptibility testing confirmed the resistance of T. asahii to amphotericin B, and in vitro pharmacodynamic results also suggest that amphotericin B is not suitable therapy for T. asahii infection. PMID- 11939584 TI - Effects of Yersinia enterocolitica O:3 derivatives on B lymphocyte activation in vivo. AB - The potential sequelae of intestinal infection with Yersinia enterocolitica include reactive arthritis, erythema nodosum, Reiter's syndrome and other autoimmune diseases. The role of the immune response in the pathogenesis of these diseases has not been fully defined, but autoimmune manifestations may be a consequence of the increase in autoantibodies as a result of polyclonal B-cell activation induced by Yersinia. We investigated the effects of Y. enterocolitica O:3 derivatives on B lymphocyte activation in vivo. Groups of five specific pathogen free (SPF) Swiss mice were inoculated with bacterial cell extract, Yersinia outermembrane proteins (Yops) or lipopolysaccharide (LPS) obtained from Y. enterocolitica O:3 and their immunoglobulin-secreting spleen cells were detected by isotype-specific protein A plaque assay. The presence of specific anti-Yersinia antibodies and autoantibodies was determined in mouse sera by ELISA. In all experiments a marked increase in the number of secretory cells of different isotypes was observed as early as the third day after inoculation. IgG and IgM anti-Yersinia antibodies were detected in the sera of all inoculated mice, and autoantibodies against myosin in the sera of those inoculated with bacterial cell extract. The sera from animals stimulated with LPS reacted with myelin, actin and laminin, while the sera from mice inoculated with Yops reacted with myelin, thyroglobulin and cardiolipin. These results suggest that SPF Swiss mice inoculated with any one of the Y. enterocolitica derivatives tested exhibited polyclonal activation of B lymphocytes as a result of stimulation by various bacterial components and not only LPS stimulation. PMID- 11939585 TI - Cells and signaling in oligodendrocyte development. PMID- 11939586 TI - Localization of the neuronal class III beta-tubulin in oligodendrogliomas: comparison with Ki-67 proliferative index and 1p/19q status. AB - The class III beta-tubulin isotype (betaIII) is widely regarded as a neuronal marker in development and neoplasia. Whereas the expression of betaIII in neuronal/neuroblastic tumors is differentiation-dependent, the aberrant expression of this cytoskeletal protein in astrocytomas is associated with an ascending gradient of malignancy. To test the generality of this observation we have compared the immunoreactivity (IR) profiles of the betaIII isotype with the Ki-67 nuclear antigen proliferative index in 41 archival, surgically excised oligodendrogliomas (32 classical [WHO grade II] and 9 anaplastic [WHO grade III]). Seventeen of 41 tumors were examined by quantitative microsatellite analysis for loss of 1p and/or 19q. Minimal deletion regions were defined on 1p (D1S468, D1S214) and 19q (D19S408, D19S867). Three of 10 classical oligodendrogliomas had combined 1p/19q loss, while 2 exhibited loss of either 1p or 19q. Three of 7 anaplastic tumors had combined 1p/19q loss. BetaIII IR was present in all tumors, but was significantly greater in the anaplastic (median labeling index [MLI] 61%, interquartile range [IQR] 55%-64%) as compared with the classical variants (MLI, 19%, IQR, 11-36%) (p < 0.0001). A highly significant relationship was found to exist between betaIII and Ki-67 LIs (betaIII, p < 0.0001 and Ki-67, p < 0.0001. r = 0.809). BetaIII localization delineated hitherto understated unipolar or bipolar tumor phenotypes with growth cones and leading cell processes resembling migrating oligodendrocyte progenitor cells. Codistribution of betaIII and GFAP IR was present in "gliofibrillary" tumor areas. Synaptophysin IR was detected in rare tumor cells (mean LI, 0.7%), and only in 4/41 samples (10%), denoting a lack of relationship between betaIII and synaptophysin expression. No significant differences in betaIII LIs were observed in tumors with 1p and/or 19q loss as compared to those with 1p/19q intact status. Increased betaIII IR in oligodendrogliomas is associated with an ascending degree of malignancy and thus is a potentially useful tumor marker. However, the significance of high betaIII LIs in low-grade oligodendrogliomas with respect to prognostic and predictive value requires further evaluation. Class III beta tubulin expression in oligodendrogliomas should not be construed as a priori evidence of divergent neuronal differentiation. PMID- 11939587 TI - Impact of genotype and morphology on the prognosis of glioblastoma. AB - The recognition of molecular subsets among glioblastomas has raised the question whether distinct mutations in glioblastoma-associated genes may serve as prognostic markers. The present study on glioblastomas (GBM) from 97 consecutively sampled adult patients is based on a clinical, histopathological, immunohistochemical, and molecular genetic analysis. Parameters assessed were age at diagnosis, survival, cell type, proliferation, necrosis, microvascular proliferation, sarcomatous growth, lymphocytic infiltration, thromboses, calcifications, GFAP expression, MIB-1 index, loss of heterozygosity (LOH) of the chromosomal arms 1p, 10p, 10q, 17p, 19q and structural alterations in the TP53, EGFR and PTEN genes. As in previous studies, younger age was significantly associated with better survival. Among the molecular parameters, TP53 mutations and LOH10q emerged as favorable and poor prognostic factors, respectively. TP53 mutations were a favorable prognostic factor independent of whether glioblastomas were primary or secondary. LOH1p or 19q, lesions suspected to be over-represented in long term survivors with malignant glioma, were not associated with better survival. However, the combination of LOH1p and LOH19q defined GBM patients with a significantly better survival. Notably, these patients did not exhibit morphological features reminiscent of oligodendroglioma. These findings indicate that genotyping of glioblastoma may provide clinical information of prognostic importance. PMID- 11939588 TI - Molecular markers that identify human astrocytomas and oligodendrogliomas. AB - The classification of human gliomas is currently based solely on neuropathological criteria. Prognostic and therapeutic parameters are dependent upon whether the tumors are deemed to be of astrocytic or oligodendroglial in origin. We sought to identify molecular reagents that might provide a more objective parameter to assist in the classification of these tumors. In order to identify mRNA transcripts for genes normally transcribed exclusively by oligodendrocytes. Northern blot analysis was carried out on RNA samples from 138 human gliomas. Transcripts encoding the myelin basic protein (MBP) were found in an equally high percentage of tumors that by neuropathological criteria were either astrocytic or oligodendroglial. In contrast, proteolipid protein (PLP) and cyclic nucleotide phosphodiesterase (CNP) mRNA molecules were found significantly more often in oligodendrogliomas than in astrocytomas. The strongest association with histological typing was found with the transcript for the myelin galactolipid biosynthetic enzyme UDP-galactose: ceramide galactosytransferase (CGT), which was about twice as frequently detected in tumors of oligodendroglial type. Results of glycolipid analyses were previously reported on a subset of the tumors studied herein. Statistical analyses of both molecular and biochemical data on this subset of astrocytomas, oligoastrocytomas, and oligodendrogliomas were performed to determine if a panel of markers could be used to separate astrocytic and oligodendroglial tumors. The presence of asialo GM1 (GA1) and the absence of paragloboside occurred most frequently in oligodendrogliomas. Ceramide monohexoside (CMH) levels correlated highly with the expression of mRNA for 4 myelin proteins: CGT, MBP, CNP, and PLP. The best combination of 2 markers of oligodendroglial tumors was CGT and GA1; the best combination of 3 markers was the presence of CGT, GA1, and the absence of paragloboside. We conclude that this combination of markers could be useful in distinguishing between astrocytic and oligodendroglial tumors. PMID- 11939589 TI - Cell type-specific expression of neuropilins in an MCA-occlusion model in mice suggests a potential role in post-ischemic brain remodeling. AB - Neuropilin-1 and -2 (NP-1/NP-2) are transmembrane receptors that play a role in axonal guidance by binding of class III semaphorins, and in angiogenesis by binding of the vascular endothelial growth factor isoform VEGF165 and placenta growth factor (PLGF). We investigated the expression pattern of NP-1/NP-2, their co-receptors, vascular endothelial growth factor receptor-1 and -2 (VEGFR-1, VEGFR-2), and their ligands, class III semaphorins, VEGF and PLGF, following experimental cerebral ischemia in mice. By means of in situ hybridization and immunohistochemistry we observed loss of expression of class III semaphorins in neurons in the infarct/peri-infarct area. In contrast, we observed high expression of NP-1 in vessels, neurons, and astrocytes surrounding the infarct. VEGF and PLGF were upregulated in different cell types following stroke. Our results suggest a shift in the balance between semaphorins and VEGF/PLGF, which compete for NP-binding. Possibly, the loss of semaphorins facilitates binding of the competing ligands (VEGF/PLGF), thus inducing angiogenesis. In addition, the observed expression patterns further suggest a neurotrophic/neuroprotective role of VEGF/PLGF. PMID- 11939590 TI - Globus pallidus glial pigment and its changes with age and chronic illness in childhood. AB - Glial lipopigment appears in the globus pallidus without accumulating in neurons (except for late adolescence) in multiple chronic childhood diseases. In this observational study, we compared the age-related development of glial pigmentation in children with the chronic illness (cystic fibrosis) and children dying acutely. A secondary goal was to search for pallidal neuronal lipopigment in childhood. We recorded pigmentation in the brains of 37 consecutive cystic fibrosis children ranging in age from 0-23 yr and in 17 controls ranging in age from 0-18 yr. We characterized the lipofuscin histochemically and used several regression models to describe the mode of deposition. We observed that in the controls, intraglial pallidal pigment accumulated in 2 forms (relatively large globules and, separately, as clusters of fine granules) at a slow rate during childhood. In cystic fibrosis, both forms of pallidal glial pigment started accumulating at a younger age and were deposited far more rapidly. There was a further increase in the rate of accumulation between 8 and 10 yr of age. We did not encounter pallidal neuronal lipofuscin at any age. These observations are consistent with 2 propositions: 1) that globus pallidus glial cells are unique in their ability to accumulate lipofuscin before it accumulates in nearby neurons; and 2) that they are particularly susceptible to some systemic effect of this chronic illness. PMID- 11939591 TI - Altered distribution of cell cycle transcriptional regulators during Alzheimer disease. AB - A number of mechanisms have been proposed to contribute to the selective neuronal cell loss observed during Alzheimer disease (AD). These include the formation and accumulation of amyloid-beta (Abeta)-containing plaques, neurofibrillary tangles (NFTs), and inflammatory processes mediated by astrocytes and microglia. Neuronal responses to such insults in AD brain include increased protein levels and immunoreactivity for kinases known to regulate cell cycle progression. One down stream target of these cell cycle regulatory proteins, the Retinoblastoma susceptibility gene product (pRb), has been shown to exhibit altered expression patterns in AD. Furthermore, in vitro studies have implicated pRb and one of the transcription factors it regulates, E2F1, in Abeta-induced cell death. To further explore the role of these proteins in AD, we examined the distribution of the E2F1 transcription factor and the hyperphosphorylated form of pRb (ppRb), which is unable to bind and regulate E2F activity, in the cortex of patients with AD and in non-demented controls. We observed increased ppRb and E2FI immunoreactivity in AD brain, with ppRb predominately located in the nucleus and E2F1 in the cytoplasm. Although neither of these proteins significantly co localized with NFTs, both ppRb and E2F1 were found in cells surrounding a subset of Abeta-containing plaques. These results support a role for G1 to S phase cell cycle regulators in AD. PMID- 11939593 TI - Ab initio conformational studies on diols and binary diol-water systems using DFT methods. Intramolecular hydrogen bonding and 1:1 complex formation with water. AB - Studies on the conformational equilibrium for the following diols, ethane-1,2 diol (12EG, CAS 107-21-1), 2R-D-(-)-propane-1,2-diol (12PG, CAS 4254-14-2), (2S,3S)-L-(+)-butane-2,3-diol (L23BD, CAS 19132-06-0), and (2S,3R)-meso-butane 2,3-diol (m23BD, CAS 5341-95-7), are described using Gaussian ab initio calculations involving density functional theory (DFT) methods. We also report in this article results on the stability and conformation for the 1:1 water-diol complex formed by ethane-1,2-diol, propane-1,2-diol, and L- and meso-butane-2,3 diol. The relative stability of the intramolecular (internal) hydrogen bond in a range of diols (n = 2 to 6), based on ab initio geometry optimization and determination of the -O...H- distance, dOH, and -O-H...O- angle, theta, increases through the sequence 1,2 approximately equals 2,3 < 1,3 < 1,4 approximately equals 1,5 approximately equals 1,6, as judged from the bond linearity and -O...H separation. Quantum mechanical and topological analysis of possible intramolecular hydrogen bonding in this complete series of diols provides convincing evidence for this in diols in which the hydroxyl groups are separated by three or more carbon atoms, that is, in (n, n+m) diols for m > or = 2, but not for ethane-1,2-diol or other vicinal diols, which do not satisfy Popelier's topological and electron density criteria based on the AIM theory of Bader. Based on these criteria it is unlikely that vicinal diols are in fact capable of forming an intramolecular hydrogen bond, in spite of geometric and spectroscopic data in the literature suggesting otherwise. PMID- 11939592 TI - The neuronal migration defect in mice with Zellweger syndrome (Pex5 knockout) is not caused by the inactivity of peroxisomal beta-oxidation. AB - The purpose of this study was to investigate whether deficient peroxisomal beta oxidation is causally involved in the neuronal migration defect observed in Pex5 knockout mice. These mice are models for Zellweger syndrome, a peroxisome biogenesis disorder. Neocortical development was evaluated in mice carrying a partial or complete defect of peroxisomal beta-oxidation at the level of the second enzyme of the pathway, namely, the hydratase-dehydrogenase multifunctional/bifunctional enzymes MFP1/L-PBE and MFP2/D-PBE. In contrast to patients with multifunctional protein 2 deficiency who present with neocortical dysgenesis, impairment of neuronal migration was not observed in the single MFP2 or in the double MFP1/MFP2 knockout mice. At birth, the double knockout pups displayed variable growth retardation and about one half of them were severely hypotonic, whereas the single MFP2 knockout animals were all normal in the perinatal period. These results indicate that in the mouse, defective peroxisomal beta-oxidation does not cause neuronal migration defects by itself. This does not exclude that the inactivity of this metabolic pathway contributes to the brain pathology in mice and patients with complete absence of functional peroxisomes. PMID- 11939594 TI - Novel computer program for fast exact calculation of accessible and molecular surface areas and average surface curvature. AB - New computer programs, SurfRace and FastSurf, perform fast calculations of the solvent accessible and molecular (solvent excluded) surface areas of macromolecules. Program SurfRace also calculates the areas of cavities inaccessible from the outside. We introduce the definition of average curvature of molecular surface and calculate average molecular surface curvatures for each atom in a structure. All surface area and curvature calculations are analytic and therefore yield exact values of these quantities. High calculation speed of this software is achieved primarily by avoiding computationally expensive mathematical procedures wherever possible and by efficient handling of surface data structures. The programs are written initially in the language C for PCs running Windows 2000/98/NT, but their code is portable to other platforms with only minor changes in input-output procedures. The algorithm is robust and does not ignore either multiplicity or degeneracy of atomic overlaps. Fast, memory-efficient and robust execution make this software attractive for applications both in computationally expensive energy minimization algorithms, such as docking or molecular dynamics simulations, and in stand-alone surface area and curvature calculations. PMID- 11939595 TI - Specific force field parameters determination for the hybrid ab initio QM/MM LSCF method. AB - The pure quantum mechanics method, called Local Self-Consistent Field (LSCF), that allows to optimize a wave function within the constraint that some predefined spinorbitals are kept frozen, is discussed. These spinorbitals can be of any shape, and their occupation numbers can be 0 or 1. Any post-Hartree-Fock method, based on the restricted or unrestricted Hartree-Fock Slater determinant, and Kohn-Sham-based DFT method are available. The LSCF method is easily applied to hybrid quantum mechanics/molecular mechanics (QM/MM) procedure where the quantum and the classical parts are covalently bonded. The complete methodology of our hybrid QM/MM scheme is detailed for studies of macromolecular systems. Not only the energy but also the gradients are derived; thus, the full geometry optimization of the whole system is feasible. We show that only specific force field parameters are needed for a correct description of the molecule, they are given for some general chemical bonds. A careful analysis of the errors induced by the use of molecular mechanics in hybrid computation show that a general procedure can be derived to obtain accurate results at low computation effort. The methodology is applied to the structure determination of the crambin protein and to Menshutkin reactions between primary amines and chloromethane. PMID- 11939596 TI - Theoretical study on the mechanism of the 1CHCl + NO reaction. AB - The complex doublet potential energy surface of the CHClNO system, including 31 minimum isomers and 84 transition states, is investigated at the QCISD(T)/6 311G(d, p)//B3LYP/6-31G(d, p) level in order to explore the possible reaction mechanism of the singlet CHCl with NO. Various possible isomerization and dissociation channels are probed. The initial association between 1CHCl and NO at the terminal N-site can almost barrierlessly lead to the chainlike adducts HClCNO a (a1, a2) followed by the direct Cl-extrusion to product P9 Cl + HCNO, which is the most feasible channel. Much less competitively, a (a1, a2) undergoes a ring closure leading to the cyclic isomer c-C(HCl)NO d followed by a concerted Cl shift and N-O cleavage of d to form the branched isomers ClNC(H)O f (f1, f2). Eventually, f (f1, f2) may take a direct H-extrusion to produce P7 H + ClNCO or a concerted 1,2-H-shift and Cl-extrusion to form P1 Cl + HNCO. The low-lying products P2 HCl + NCO, P3 Cl + HOCN, P14 HCO + 3NCl, P6 ClO + HCN, and P13 ClNC + OH may have the lowest yields observed. Our calculations show that the product distributions of the title reaction are quite different from those of the analogous 1CHF + NO reaction, yet are similar to those of another analogous 3CH2 + NO reaction. The similarities and discrepancies among the three reactions are discussed in terms of the substitution effect. The present article may assist in future experimental identification of the product distributions for the title reaction and may be helpful for understanding the halogenated carbene chemistry. PMID- 11939597 TI - Conformational analysis of the tyrosine dipeptide analogue in the gas phase and in aqueous solution by a density functional/continuum solvent model. AB - The conformational behavior of a dipeptide analogue of tyrosine (TDA) has been investigated by density functional methods using the polarizable continuum model (PCM) for the description of solvent effects. Our study points out the interplay of backbone and side chain contributions in determining the relative stabilities of energy minima. In particular, stabilizing interactions between the NH bond and the aromatic ring have a significant effect. The topology of the potential energy surface is significantly modified in aqueous solution due to a general widening of low energy regions and to a stabilization of helical structures. PMID- 11939598 TI - Density functional theory for efficient ab initio molecular dynamics simulations in solution. AB - We present a density functional for first-principles molecular dynamics simulations that includes the electrostatic effects of a continuous dielectric medium. It allows for numerical simulations of molecules in solution in a model polar solvent. We propose a smooth dielectric model function to model solvation into water and demonstrate its good numerical properties for total energy calculations and constant energy molecular dynamics. PMID- 11939599 TI - Computation of pressure components due to Class II force fields. AB - It has been shown that purely angle dependent terms in a force field do not contribute to the total pressure in a molecular simulation. However, this is not the case for the individual components of the pressure tensor, and is also untrue for crossterms in the force field including bond stretch. In this article, we show that virial contributions to the pressure tensor are easily computed in terms of bond distance vectors and atom forces for the bond topologies present in a Class II force field. Results from a simulation of a phospholipid biomembrane using the cff97 force field show that angle and torsional crossterms make a significant contribution to the pressure tensor. PMID- 11939600 TI - Multiple grid methods for classical molecular dynamics. AB - Presented in the context of classical molecular mechanics and dynamics are multilevel summation methods for the fast calculation of energies/forces for pairwise interactions, which are based on the hierarchical interpolation of interaction potentials on multiple grids. The concepts and details underlying multigrid interpolation are described. For integration of molecular dynamics the use of different time steps for different interactions allows longer time steps for many of the interactions, and this can be combined with multiple grids in space. Comparison is made to the fast multipole method, and evidence is presented suggesting that for molecular simulations multigrid methods may be superior to the fast multipole method and other tree methods. PMID- 11939601 TI - Intravenous immunoglobulin: appropriate indications and uses in hematopoietic stem cell transplantation. AB - Intravenous immune globulin (IVIG) therapy has been prescribed in many different disease states. Hyperimmune products are also available. Recently, routine use for many indications has come under scrutiny secondary to high cost, limited supply, and unclear benefit. IVIG is U.S. Food and Drug Administration-approved for application in hematopoietic stem cell transplantation (HSCT), a very common indication for its use. In an attempt to clarify the most appropriate indications and doses in HSCT recipients, we conducted a MEDLINE search in which we reviewed all relevant articles from 1966 to the present. Search terms included bone marrow transplantation, intravenous immune globulin, hyperimmune globulin, GVHD, and cytomegalovirus (CMV). Also, the references of all pertinent studies and review articles were scanned for studies missed via MEDLINE. CMV prophylaxis/treatment and GVHD prophylaxis are the 2 indications with the most significant clinical support, but there are very few prospective, randomized, controlled trials reported. Furthermore, sample size usually was small, included heterogeneous patient populations, and employed different primary end points. Several reports support IVIG therapy in combination with ganciclovir for prevention and treatment of CMV infection, whereas others have shown ganciclovir monotherapy to be effective, blurring the benefit of IVIG administration. CMV IgG data are also imprecise and difficult to interpret. The role of IVIG therapy in prevention and treatment of GVHD also is vague. Only 1 randomized investigation showed a benefit in the prevention of acute GVHD, and no studies showed efficacy in chronic GVHD prophylaxis and therapy. Reports examining the utility of IVIG or CMV IgG in HSCT are hampered by marked variation in trial design and dosing and diverse patient characteristics. Although IVIG may be useful as a component of preemptive therapy and treatment of CMV disease, its contribution to the prevention of reactivation of CMV infection is dubious. Extended IVIG therapy during GVHD prevention may impair recovery of humoral immunity, and its role in prophylaxis and therapy of GVHD has not been clearly defined. Hospital monitoring programs may be a valuable way to detect areas of high use and allow for streamlining of prescribing. PMID- 11939602 TI - Posttransplantation cyclophosphamide facilitates engraftment of major histocompatibility complex-identical allogeneic marrow in mice conditioned with low-dose total body irradiation. AB - Cyclophosphamide (Cy) has been studied extensively for its immunosuppressive properties and is frequently combined with total body irradiation (TBI) as conditioning prior to HLA-identical allogeneic blood or marrow transplantation (alloBMT) in humans. Because Cy is most effective at suppressing host-versus graft reactions when the drug is given after the transplantation (Mayumi H et al. Transplant Proc. 1986;18:363-369), we investigated whether posttransplantation Cy could prevent rejection of allogeneic marrow in mice conditioned with low-dose TBI. In a mouse model, posttransplantation Cy reduced the dose of TBI required from 500 cGy to < or = 200 cGy for the engraftment of 10 million major histocompatibility complex (MHC)-identical marrow cells in 100% of recipients. In animals conditioned with low-dose TBI and posttransplantation Cy, donor chimerism was proportional to the dose of TBI, was present in multiple hematopoietic lineages, and was associated with the indefinite survival of donor-strain skin grafts. In contrast, animals conditioned with either TBI alone or posttransplantation Cy alone failed to achieve engraftment after alloBMT and contained antidonor cytotoxic T-cells. Although <5% donor chimerism could be induced without TBI by transplanting > or = 50 million MHC-identical cells and administering posttransplantation Cy, the addition of low-dose TBI reduced the dose of donor cells required for alloengraftment and increased long-term donor chimerism to >50%. These data demonstrate that low-dose TBI and posttransplantation Cy cooperate to prevent graft rejection following the transplantation of standard doses of MHC-identical marrow cells. PMID- 11939603 TI - Effect of total body irradiation dose escalation on outcome following T-cell depleted allogeneic bone marrow transplantation. AB - Prior studies of non-T-cell-depleted (TCD) transplantation have demonstrated a reduction in relapse in patients receiving escalated doses of TBI; however, overall survival in these studies was not significantly improved due to increased treatment-related toxicity seen at the higher doses of irradiation. Toxicity was in part related to an increased incidence of GVHD. Because T-cell depletion of donor bone marrow reduces the incidence of GVHD and other treatment-related complications after allogeneic bone marrow transplantation, it was postulated that TBI dose may be safely escalated in this setting and may decrease the risk of relapse following TCD BMT. Herein, we report the results of a trial assessing the safety and impact of escalated doses of TBI after TCD BMT. Two hundred adults with hematologic malignancies were treated in consecutive cohorts defined by increasing doses of TBI (1400, 1480, and 1560 cGy) in combination with cyclophosphamide. In vitro T-cell depletion using anti-CD6 monoclonal antibody was used for GVHD prophylaxis. The incidence of grade II or greater acute GVHD in patients receiving 1560 cGy (36%) was significantly higher than in patients receiving 1400 cGy (18%) (P = .04) or 1480 cGy (13%) (P = .01). Two-year treatment-related mortality was significantly higher in patients who received 1560 cGy of TBI (33%) than in those who received 1400 cGy (20%) (P = .04) or 1480 cGy (19%) (P = .05). The increased dose of TBI did not reduce the rates of relapse, with the estimated 2-year risk of relapse being 24% (1400 cGy), 24% (1480 cGy), and 31% (1560 cGy) for the 3 cohorts of patients. Overall survival at 2 years was inferior for patients receiving 1560 cGy of TBI (36%) compared with those who received 1400 cGy (55%) or 1480 cGy (58%) (P = .01). We conclude that dose escalation of TBI is associated with increased GVHD and inferior survival following TCD BMT. Future efforts to reduce the risk of relapse after TCD BMT should focus on immunologic methods to induce the graft-versus-leukemia effect after BMT rather than intensification of the ablative regimen by escalation of irradiation dose. PMID- 11939604 TI - Conditioning therapy with intravenous busulfan and cyclophosphamide (IV BuCy2) for hematologic malignancies prior to allogeneic stem cell transplantation: a phase II study. AB - Busulfan (Bu) is commonly used as a component of conditioning regimens for hematopoietic stem cell transplantation. Precise delivery of the oral formulation is compromised by erratic gastrointestinal absorption. An IV Bu formulation was developed to provide dose assurance and complete bioavailability. In a phase I study, the plasma bioequivalence of IV Bu was established at approximately 80% of the oral dose. We now report the findings of the first phase II study, in which 61 adults with hematologic cancers were treated with a Bu-cyclophosphamide (BuCy) regimen consisting of IV Bu (0.8 mg/kg every 6 hours x 16) followed by Cy (60 mg/kg qd x 2) and transplantation of stem cells from an HLA-matched sibling donor. The median age of study participants was 37 years; 75% of patients had active disease; 48% were heavily pretreated, and 13% had undergone a prior transplantation. Median follow-up was 2.3 years; median time to engraftment (absolute neutrophil count, >0.5 x 10(9)/L) was 13 days; 100% of patients with cytogenetic and/or molecular markers had documented chimerism; and there were no engraftment failures. Two-year overall and disease-free survival were 67% and 42%, respectively. There were no unexpected toxic reactions. Fatal veno-occlusive disease occurred in 2 patients, 1 of whom had undergone a prior transplantation. Treatment-related mortality at 100 days was 9.8% (6/61). Bu pharmacokinetics after IV drug administration demonstrated high inter- and intrapatient consistency; 86% of patients maintained an area under the curve between 800 and 1500 microMol-min. In conclusion, the IV Bu in this regimen was very well tolerated and demonstrated excellent antitumor efficacy, most likely because of dose assurance with predictable pharmacokinetics. PMID- 11939605 TI - Poor outcome in steroid-refractory graft-versus-host disease with antithymocyte globulin treatment. AB - Treatment of acute graft-versus-host disease (aGVHD) has relied on high-dose steroids, but less than 50% of patients show durable remission. Antithymocyte globulin (ATG) has become a standard salvage therapy. We now report our experience with ATG for the treatment of steroid-refractory GVHD in 69 patients treated from January 1, 1980, to May 1, 1999. Patients with GVHD were given an overall grade using standard criteria. Overall responses were similar to those in previously published literature. However, long-term survival for this group of patients was dismal. Of the 69 patients treated with ATG for steroid-refractory GVHD, only 3 (5%) are currently alive. The median survival of these patients by GVHD grade was 4.1 months for grade 2, 3.6 months for grade 3, and 2.7 months for grade 4. The age range of the surviving patients was 3 to 25 years. Only 5% of the deaths were due to relapse, with the remaining deaths due to GVHD, infection, and/or organ failure. In conclusion, ATG treatment can produce objective responses in patients with aGVHD, but these responses do not result in long-term survival. Given the poor survival rates of patients treated with ATG for steroid refractory GVHD, treatment with ATG as standard therapy should be reconsidered. Patients with steroid-refractory GVHD should be enrolled in clinical study until there are data to support a standard salvage therapy. PMID- 11939606 TI - Hematopoietic stem cell transplantation for advanced myelodysplastic syndrome after conditioning with busulfan and fractionated total body irradiation is associated with low relapse rate but considerable nonrelapse mortality. AB - The objectives of this study were to develop transplantation regimens for patients with advanced myelodysplastic syndrome (MDS) that would be associated with low transplantation-related mortality and improved relapse-free survival. Sixty patients with advanced MDS or acute myeloid leukemia evolving from MDS (sAML), 12 to 62 years old (median, 40 years), were conditioned with busulfan (7 mg/kg) and TBI (6 x 200 cGy) (BU/TBI) and received transplants from related (n = 20) or unrelated donors (n = 40). By French-American-British (FAB) criteria, 21 patients had refractory anemia with excess blasts (RAEB), 16 had RAEB in transformation (RAEB-T), 15 had sAML, and 8 had chronic myelomonocytic leukemia (CMML). By International Prognostic Scoring System (IPSS) criteria, 1 patient had low, 10 had intermediate-1, 13 had intermediate-2, and 31 had high-risk MDS (5 patients had proliferative CMML). All evaluable patients achieved sustained engraftment. The cumulative incidence (CI) of acute GVHD grades II to IV was 83% with unrelated donors and 85% with related donors. The CI of relapse was 25% at 3 years. The incidence of nonrelapse mortality (NRM) at 100 days was 38%. The Kaplan-Meier estimate of survival was 26% at 3 years. Major causes of death were relapse, organ failure, GVHD, and infection. In multivariate analysis, improved relapse-free survival was associated with good cytogenetic risk (P = .002) and shorter disease duration (P = .004). NRM was increased with longer disease duration (P = .0002), positive cytomegalovirus serology (P = .02), and male sex (P = .02). Relapse was associated with poor cytogenetic risk (P = .0004). Thus, BU/TBI conditioning as used in this trial was associated with relapse rates comparable to those observed with a previously used more intensive regimen combining BU/TBI with cyclophosphamide. However, despite the omission of cyclophosphamide, transplantation-related morbidity and mortality were considerable, particularly with transplants from unrelated donors. Future trials should explore the efficacy and tolerability of reduced-intensity conditioning regimens. PMID- 11939607 TI - Re: measurement of chimerism after hematopoietic stem cell transplantation. PMID- 11939608 TI - Increased levels of transforming growth factor-beta1 in essential hypertension. AB - BACKGROUND: Transforming growth factor-beta (TGF-beta) is a multifunctional cytokine that has been linked to vascular remodeling processes, myocardial hypertrophy, and renal fibrosis. Recently a correlation between serum levels of TGF-beta1 and blood pressure (BP) levels in patients with end-stage renal disease was shown. In addition, it is not clear whether TGF-beta1 is a causative factor in the pathogenesis of essential hypertension and associated with hypertensive target organ damage (TOD). METHODS: Using a TGF-beta1-specific sandwich ELISA, we compared plasma levels of active and total TGF-beta1 of 30 normotensive persons and 85 patients with essential hypertension with and without TOD, as measured by microalbuminuria or left ventricular hypertrophy. RESULTS: Active and total TGF beta1 levels were significantly higher in plasma of patients with essential hypertension than in normotensive controls (P < .05 and P < .01, respectively). However, neither active nor total TGF-beta1 correlated with systolic or diastolic BP (R2 < 0.14 for all parameters). Levels of active and total TGF-beta1 were significantly higher in hypertensive patients with than without TOD (P < .05). CONCLUSIONS: Active and latent TGF-beta1 levels are markedly increased in plasma of hypertensive patients. We assume that TGF-beta1 contributes substantially to the development of TOD in essential hypertension, independent of BP levels. PMID- 11939609 TI - Renal medullary blood flow and salt load in Lyon hypertensive rats. AB - The present work studied renal medullary blood flow (MBF) and its response to salt load in Lyon hypertensive (LH) rats to understand the mechanisms underlying the abnormal renal sodium excretion exhibited by LH rats. Experiments were conducted in uninephrectomized, anesthetized, and volume-expanded 15-week-old male LH and their normotensive (LL) controls. Under standard diet, LH rats exhibited a blunted pressure diuresis and natriuresis associated with an absence of pressure-induced increase in MBF compared to LL rats. One week of salt load (2% NaCl as drinking water) induced a significant increase in blood pressure (BP) in LH (+11 mm Hg) than in LL (+6 mm Hg) rats associated with a decrease in MBF in LH rats only (from 182 +/- 25 to 122 +/- 20 perfusion units, P < .001). Finally, despite the salt load-induced increase in pressure natriuresis, it remained significantly lower in LH than in LL rats. The results show an alteration in MBF regulation in LH rats and suggest that this abnormality may be involved in their blunted pressure natriuresis and their enhanced salt sensitivity. PMID- 11939610 TI - Sexual activity and plasma testosterone levels in hypertensive males. AB - The aim of this study was to compare sexual activity and plasma testosterone levels of hypertensive men with those of healthy normotensive controls. We investigated 110 newly diagnosed, never treated hypertensive (blood pressure [BP] > or = 140/95 mm Hg) men and 110 healthy normotensive (diastolic BP <90 mm Hg) men. All of them were aged 40 to 49 years, married, without any previous sexual dysfunction, nondiabetic, nonobese (body mass index <28 kg/m2), nonsmoking, and not taking any drug. All subjects were evaluated in the morning after an overnight fast. Clinical evaluation included BP, body weight, and height measurements, determination of testosterone, and an interview about sexual activity, assessed as number of sexual intercourse episodes per month. Hypertensive men presented a 25% reduction in sexual activity as compared to normotensive men (5.9 +/- 2.6 v 7.9 +/- 2.5 sexual intercourse episodes per month, respectively, P < .01) and a 12% reduction in testosterone levels (510.6 +/- 151.9 ng/dL v 578.6 +/- 146.8 ng/dL, P < .01). In both normotensive and hypertensive men Pearson's correlation analysis showed a significant positive correlation between testosterone levels and sexual activity and a significant negative correlation between testosterone and age and between testosterone and BP values. Multiple regression analysis confirmed a significant inverse relationship between testosterone and age in normotensive men, whereas only a nonsignificant trend was found in the hypertensive ones. In addition, a significant inverse correlation between testosterone and BP levels was confirmed in hypertensive men limited to systolic BP, whereas a nonsignificant trend was observed in the normotensive controls. In conclusion, these findings suggest a relationship between essential hypertension and impaired testosterone levels in men. The elucidation of the nature of such a relationship and its physiologic and clinical significance needs further investigation. PMID- 11939611 TI - Dietary sodium and target organ damage in essential hypertension. AB - In addition to its widely contested influence on arterial pressure, dietary sodium may exert some nonpressure-related effects on left ventricular mass in humans. In the present study, we hypothesized that sodium intake (estimated by two consecutive measurements of 24-h urinary sodium excretion) may amplify the effect of arterial pressure on target organ damage (ie, left ventricular mass and microalbuminuria) in a large group of normotensive subjects and patients with never-treated uncomplicated essential hypertension. Left ventricular mass (M-mode echocardiography) and urinary albumin excretion were assessed in 839 subjects (471 men and 368 women) aged 15 to 70 years, with elevated (60%) or normal arterial pressure. In the entire population, multivariate analysis indicated that the relationship between urinary sodium excretion and left ventricular mass index (beta = 0.02, P < .01) as well as urinary albumin excretion (beta = 0.001, P < .0001) was independent from sex, age, body mass index, and systolic arterial pressure. When subjects were divided into quintiles according to urinary sodium excretion, left ventricular mass index and urinary albumin excretion increased significantly from the lowest to the highest quintile in both genders, despite similar values of systolic arterial pressure. The slope of the regression line linking systolic arterial pressure to left ventricular mass index (in men) and urinary albumin excretion (in the entire population) obtained within each quintile of urinary sodium excretion, progressively and linearly increased from the lowest to the highest quintile. These results suggest that sodium intake may amplify the effect of arterial pressure on both the left ventricle and the kidney, and thus suggest that dietary sodium may be an independent factor of cardiovascular risk. PMID- 11939613 TI - Large and small artery compliance changes during hemodialysis. AB - BACKGROUND: Cardiovascular disease is the major cause of mortality and morbidity in patients undergoing hemodialysis. Reduced arterial compliance is an independent predictor of cardiovascular mortality in end stage renal disease and can be measured noninvasively using pulse wave analysis technology. METHODS: Ten chronic hemodialysis patients were evaluated using pulse wave analysis to determine large and small vessel compliance before hemodialysis, midway through the treatment, and at the end of the treatment. Serum calcium was measured before and after hemodialysis. RESULTS: No significant changes in systolic blood pressure (BP), diastolic BP, pulse pressure, or heart rate occurred during hemodialysis. A significant decrease in small vessel compliance (C2) occurred during the treatment, with the mean C2 decreasing from 5.6 +/- 2.7 mL/mm Hg x 10 predialysis to 3.3 +/- 1.5 mL/mm Hg x 10 midway through the treatment (P = .04) and to 3.9 +/- 4.3 mL/mm Hg x 10 at the end of the dialysis treatment. There was no significant change in large vessel compliance (C1). Serum calcium increased significantly during the dialysis treatment from 8.0 +/- 1.2 mg/dL to 9.8 +/- 0.9 mg/dL (P = .003). CONCLUSIONS: Significant decreases in small artery compliance occur without systolic or diastolic BP changes during routine hemodialysis. A significant increase in serum calcium also occurred during hemodialysis. Measurements of arterial compliance during hemodialysis may be an important tool to identify patients with vascular responses, which may place them at greater risk for cardiovascular disease. PMID- 11939612 TI - Inhibition of inducible nitric oxide synthase during high dietary salt intake. AB - BACKGROUND: Previous studies indicate that nitric oxide (NO) is involved in the regulation of blood pressure (BP) and natriuresis in response to high sodium intake. We investigated the role of inducible NO synthase (iNOS) in response to an increased salt intake. METHODS: Conscious, chronically catheterized rats were exposed to a high-salt (6%) diet for 14 days while receiving vehicle or aminoguanidine ([AG]; 250 mg/kg/24 h), which selectively inhibits iNOS. A group of rats on normal salt intake + AG were also studied. RESULTS: Aminoguanidine had no impact on BP (120 +/- 2 v 116 +/- 1 mm Hg, control v day 14) or 24-h urinary nitrite and nitrate excretion (UNOxV), in rats on normal salt but prevented lipopolysaccharide-induced hypotension. High salt alone had no impact on BP (120 +/- 1 v 121 +/- 1 mm Hg), whereas UNaV (1.3 +/- 0.2 v 3.5 +/- 0.6 microeq/min, P < .001) and UNOxV increased with high salt intake. The natriuretic response persisted (1.5 +/- 0.2 v 4.3 +/- 0.8 microeq/min, P < .005), but the increase in UNOXV was prevented with chronic AG although BP fell slightly (121 +/- 1 v 115 +/ 1 mm Hg, P < .05). There was no change in plasma volume with high salt, and 24-h UNaV increased appropriately in the presence of AG. The in vitro NOS activity was not increased in kidney homogenates by high salt diet, nor was it affected by chronic AG treatment. CONCLUSION: We conclude that NO from an iNOS source is not essential for the regulation of sodium excretion and BP in the presence of a high salt diet in a normal rat. PMID- 11939614 TI - Divergent effect of acute and chronic alcohol on arterial stiffness. AB - To study the effects of alcohol on large artery function we measured arterial wave reflection in the aorta as augmentation index (AI%) by applanation tonometry in 324 subjects (18 to 86 years, 223 male). In eight subjects, when ingested acutely, red wine containing alcohol (0.8 g/kg), but not dealcoholized wine, reduced (P < .01) blood pressure (BP), pulse wave velocity, and AI%. Men with an excessive alcohol consumption (>21 units/week) had a higher AI% (12 +/- 2 v 5 +/- 2, P < .05) and BP, particularly aortic systolic, than did those with a lesser intake. This study suggests that alcohol when ingested acutely may reduce arterial stiffness, although alcohol when ingested chronically, in excess, increases it. PMID- 11939615 TI - Pulse pressure and diurnal blood pressure variation: association with micro- and macrovascular complications in type 2 diabetes. AB - BACKGROUND: In nondiabetic subjects pulse pressure (PP) is an independent predictor of cardiovascular disease and microalbuminuria. Reduced circadian blood pressure (BP) variation is a potential risk factor for the development of diabetic complications. We investigated the association between retinopathy, nephropathy, macrovascular disease, PP, and diurnal BP variation in a group of type 2 diabetic patients. METHODS: In 80 type 2 diabetic patients we performed 24 h ambulatory BP (AMBP) and fundus photographs. Urinary albumin excretion was evaluated by urinary albumin/creatinine ratio. Presence or absence of macrovascular disease was assessed by an independent physician. RESULTS: Forty nine patients had no detectable retinal changes (grade 1), 13 had grade 2 retinopathy, and 18 had more advanced retinopathy (grades 3-6). Compared to patients without retinopathy (grade 1), patients with grades 2 and 3-6 had higher PP and blunted diurnal BP variation: night PP 55 +/- 10 mm Hg, 64 +/- 10 mm Hg, 61 +/- 15 mm Hg, P < .05 and systolic night/day ratio 89.3% +/- 7%, 94.6% +/- 8%, and 92.0% +/- 6%, P < .05 (grade 1, 2, and 3-6, respectively). Comparing nephropathy groups (45 normo-, 19 micro-, and 15 macroalbuminuric patients) results were similar: night PP 54 +/- 9 mm Hg, 57 +/- 10 mm Hg, and 70 +/- 15 mm Hg, P < .001 and systolic night/day ratio 88.9% +/- 7%, 92.0% +/- 7%, and 94.9% +/- 7%, P < .02. Likewise, compared to patients without macrovascular disease (n = 55), patients with this complication (n = 25) had higher AMBP values: night PP 57 +/- 12 mm Hg v 63 +/- 11 mm Hg, P < .05 and systolic night/day ratio 89.2% +/- 6% v 94.1% +/- 9%, P < .01. CONCLUSIONS: Increased PP and blunted diurnal BP variation are hemodynamic abnormalities associated with micro- and macrovascular complications in type 2 diabetes. PMID- 11939616 TI - Effect of continuous positive airway pressure therapy on 24-hour blood pressure in patients with obstructive sleep apnea syndrome. AB - BACKGROUND: Patients with obstructive sleep apnea syndrome (OSAS) are subject to an increased cardiovascular morbidity including systemic hypertension. Little is known about the effects of treatment with nasal continuous positive airway pressure (CPAP) on systemic hypertension. METHODS: Automated ambulatory 24-h blood pressure (BP) monitoring was performed in 88 consecutive patients who were referred for evaluation of snoring or suspected OSAS. In addition, the long-term effects of CPAP therapy on 24-h BP were assessed. RESULTS: A total of 62 patients had OSAS and 26 habitual snoring. Patients with OSAS had significantly higher mean arterial BP values than snorers (102.7 +/- 10.7 v 94.0 +/- 10.2 mm Hg; P < .01). Multiple stepwise linear regression analysis disclosed that the degree of systemic hypertension was independently associated with the severity of OSAS as determined by the apnea/hypopnea index (R = 0.43; P < .001), but not with age, body mass index, or smoking habits. Of the 62 patients with OSAS, 52 were treated with CPAP and reevaluated after 9 months. The CPAP resulted in a significant decrease in mean arterial BP (from 103.7 +/- 10.4 to 99.1 +/- 10.8 mm Hg; P < .05). For those patients with systemic hypertension whose BP improved with CPAP therapy, 24-h mean pulse pressure at baseline (r = -0.36; P < .05) as well as average heart rate during the day (r = -0.35; P < .05) turned out as predictors. CONCLUSIONS: Obstructive sleep apnea syndrome contributes, at least in part, to the development of systemic hypertension, and CPAP may improve BP values in treated OSAS patients. Predictors of a beneficial CPAP effect on BP are a high heart rate and a high pulse pressure before treatment. PMID- 11939617 TI - A "gender blind" relationship of lean body mass and blood pressure in the Tecumseh study. AB - BACKGROUND: Body size correlates positively with blood pressure (BP) but there is controversy about the roles of obesity versus muscularity in this relationship. METHODS: We examined the BP relationship with overweight, lean body mass (LBM), and muscle performance in 231 adolescents (17.25 +/- 3.07 years, 123 males). The skinfold thickness (SKINT) was used to measure overweight, as this was a growing population. RESULTS: Maximal foot torque, a measure of muscle strength, correlated strongly (r = 0.51, P < .001) to LBM attesting to the validity of the calculated LBM. Anthropometric measurements were available also in 944 adults (29.9 +/- 5.5 years, 461 men). Correlations of LBM to systolic (adolescents r = 0.52, adults r = 0.19, both P < .001) and diastolic (adolescents r = 0.47, adults r = 0.20, both P < .001) BP were highly significant. SKINT also correlated significantly to systolic and diastolic BP in adolescents and in adults, respectively. In both genders and populations an increasing SKINT was associated with a similar increase in BP, but this effect was superimposed on an average 10 mm Hg between-gender BP difference. The LBM in both groups and genders related to the BP in an identical fashion; the men were on the high and the women on the low end of the same BP/LBM correlation line. Thus, the amount of LBM erased categoric BP differences between the genders. CONCLUSIONS: The gender-related BP differences appear to reflect the inherent gender differences in muscle bulk. PMID- 11939618 TI - The forgotten Korotkoff phases: how often are phases II and III Present, and how do they relate to the other Korotkoff phases? AB - BACKGROUND: There are no data on the pattern of the Korotkoff phases in the normal population. This study was designed to describe the pattern of Korotkoff phase distribution in adults and children; to measure the duration of each of the phases; and to describe the differences between adults and children. METHODS: A total of 57 children (7 to 8 years old) and 59 adults (median age 47 years, range 30 to 62 years) were studied. The pressure in the arm cuff was deflated using a device to provide a consistent deflation rate. The Korotkoff sounds were recorded to MiniDisc from the bell of a stethoscope and each sound described as phase I, II, III, or IV. RESULTS: The most common pattern of Korotkoff phase distribution was for all five phases to be present (children [23/57; 40%], adults [24/59; 41%]). Phases I and IV were more common in children than in adults (56/57 [98%] v 47/59 [80%]; P = .002 for phase I; 52/57 [91%] v 44/59 [75%]; P = .018 for phase IV). Phases II and III were less common in children than in adults (32/57 [56%] v 50/59 [85%], P = .001 for phase II; 27/57 [47%] v 45/59 [76%], P = .001 for phase III). Phases I and IV were longer in children (median 3.9 [interquartile range, IQR 2.1 to 6.7] and 6.7 [IQR 3.2 to 9.8] sec, respectively) compared with adults (1.3 [IQR 0.7 to 2.7] and 1.7 [IQR 0.3 to 2.6), P < .001). CONCLUSIONS: There are clear differences in the Korotkoff phases between adults and children. The length of phases II and III increase with age with concomitant decrease in phases I and IV. These differences between adults and children remain unexplained. PMID- 11939619 TI - Plasma tissue inhibitor of metalloproteinase-1 levels are elevated in essential hypertension and related to left ventricular hypertrophy. AB - BACKGROUND: Essential hypertensive patients have an increased heart and arterial collagen concentration. Increased collagen synthesis can be assessed using procollagen III N peptide (PIIINP) and reduced collagen degradation measured using tissue inhibitor of metalloproteinase-1 (TIMP-1). METHODS: Plasma TIMP-1 and PIIINP levels were measured in 31 patients with essential hypertension and in 17 normotensive control subjects. The hypertensive patients were either treatment naive (n = 18) or had been without treatment for 1 month (n = 13). Both groups of patients were screened to exclude other fibrotic diseases. RESULTS: In the hypertensive patients, TIMP-1 levels were significantly (P < .0002) elevated (median 380 ng/ mL, range 160 to 1,560 ng/mL) compared with those of the normotensive control subjects (median 178 ng/mL, range 99 to 330 ng/mL). In hypertensive subjects who had never received antihypertensive therapy there were significant correlations between TIMP-1 and left ventricular posterior wall thickness in diastole (LVPWd) (r = 0.58) (P < .02) and left ventricular mass index (r = 0.58) (P < .02). There was no difference in PIIINP levels (mean +/- 2 SD) between the hypertensive (0.56 U/mL +/- 0.3) and normotensive groups (0.52 U/mL +/- 0.2). CONCLUSIONS: The increased tissue collagen III levels found in the heart and vessels of hypertensive patients is due to a reduction in collagen degradation because of high TIMP-1 levels, rather than an increase in synthesis of collagen type III. The tissue source of this TIMP-1 is unclear. PMID- 11939620 TI - Insulin increases NADH/NAD+ redox state, which stimulates guanylate cyclase in vascular smooth muscle. AB - BACKGROUND: Insulin inhibits contraction and migration of primary confluent, cultured canine vascular smooth muscle cells (VSMCs) with inducible nitric oxide synthase (iNOS) by stimulating cyclic GMP (cGMP) production. The present study was performed to determine how insulin stimulates guanylate cyclase activity in these cells. METHODS: Primary cultured VSMC were obtained from canine femoral arteries. Lactate and pyruvate levels were measured by enzymatic assays, cGMP production by radioimmunoassay, iNOS activity by conversion of arginine to citrulline, and cell contraction by photomicroscopy. RESULTS: Insulin (1 nmol/L) increased cGMP production fivefold in VSMC with iNOS while raising the lactate-to pyruvate ratio (LPR) from 3.1 +/- 0.5 to 10.0 +/- 1.6 (P < .05), indicating a rise in the ratio of reduced/oxidized nicotinamide adenine dinucleotide (NADH/NAD+) redox state of the cell. Insulin's stimulation of cGMP production was blocked by 0.1 mmol/L NG-monomethyl-L-arginine (L-NMMA) indicating dependence on iNOS activity, but insulin did not affect iNOS activity. Blocking insulin's increase in LPR by pyruvate (0.5 mmol/L) or oxaloacetate (0.5 mmol/L) completely inhibited the insulin-stimulated component of cGMP production. Pyruvate also blocked insulin's inhibition of serotonin-induced contraction in nonproliferated cells. In the absence of insulin, 5 mmol/L lactate or isocitrate increased the LPR by 420% +/- 47% and 167% +/- 20%, respectively (both P < .05), and stimulated cGMP production by 1,045% +/- 272% and 278% +/- 33%, respectively (both P < .05) by an L-NMMA-inhibitable mechanism. Although cGMP production in cells with iNOS was increased by insulin, the stimulation of cGMP production in cells without iNOS by 3-(5'-hydroxymethyl-2'furyl)-1-benzyl indazole (YC-1) was not affected by insulin, suggesting that insulin does not stimulate guanylate cyclase activity directly. CONCLUSION: We conclude that insulin increases cGMP production in VSMC with iNOS by raising the cell NADH/NAD+ redox state, which may increase the availability of iNOS-derived NO. PMID- 11939621 TI - Tissue activity of circulating prorenin. AB - Both renin and its biosynthetic precursor, prorenin, are secreted into the circulation of mammals. Although the circulating levels of prorenin can exceed those of renin by as much as 100-fold in certain conditions, there is no evidence that prorenin contributes to the synthesis of circulating angiotensin peptide synthesis or increased blood pressure (BP). In the current study, we have generated a transgenic mouse line that overexpresses human prorenin in the liver and have mated these mice to a second mouse line expressing human angiotensinogen in the heart. Double-transgenic progeny display a selective increase in angiotensin I content in the heart (but not the plasma) as compared to the single transgenic mice. These results are consistent with a model in which circulating prorenin is taken up by tissues where it can contribute to the local synthesis of angiotensin peptides. This finding may explain some of the pathologies associated with high circulating prorenin levels. PMID- 11939623 TI - The state of the art for chemometrics in analytical chemistry. PMID- 11939624 TI - Simple PVC-PPy electrode for pH measurement and titrations. AB - Cobaltabis(dicarbollide) [3,3'-Co(1,2-C2B9H11)](-)-doped polypyrrole (PPy) films have been prepared galvanostatically on glassy carbon electrodes in acetonitrile solution. The potential response behavior of the film of this new material has been investigated in some common pH buffers and in acid-base titrations. The potentiometric characteristics of the resulting films are indicative of a quasi Nernstian response (approximately 50 mV/pH unit), a linearity range from pH 12 to 3 and correlation coefficients (r2) of approximately 0.98. The electrode is suitable for pH measurements and for monoprotic titrations of strong alkalis with strong acids, and weak bases with strong acids, but the long response time hinders the use of this electrode for multiprotic titrations. The time response has been dramatically improved by reducing the film thickness by using the template effect of a non-conducting polymer (PVC) cast over the graphite surface before PPy deposition. PPy polymerization occurs in the free channels of PVC leading to the formation of PPy wires. The morphological change of PPy does not affect the slope or linearity range. The response of the PVC-PPy electrochemical sensor is rapid and the sensor is easy to prepare, at low cost, and its performance is comparable with that of commercial glass electrodes. PMID- 11939622 TI - Comparative effects of candesartan and enalapril on augmented vasoconstrictive responses to endothelin-1 in coronary vessels of spontaneously hypertensive rats. AB - BACKGROUND: The aim of this study was to compare the effect of angiotensin type-1 receptor blockade (ARB) on augmented vasoconstrictive response to endothelin-1 (ET-1) in coronary vessels of hypertensive hearts with angiotensin converting enzyme (ACE) inhibitor, candesartan cilexetil (CAN) or enalapril was administered for 3 weeks in spontaneously hypertensive rats (SHR). METHODS: We used SHR (9 to 12 weeks old, n = 18) and Wistar-Kyoto (WKY) rats (n = 6). Systolic blood pressure was measured once a week. Spontaneously hypertensive rats were divided into three groups. Enalapril malate (10 mg/day) or CAN (10 mg/day) was administered orally in each of six SHR in each group receiving treatment for 3 weeks. The control group (n = 6) received no treatment. At the end of this experiment, the hearts were isolated. Isolated hearts mounted on a Langendorff apparatus after weighing were then perfused with modified Krebs-Henseleit buffer at constant pressure (75 mm Hg). The coronary perfusion pressure and coronary flow were measured during perfusion of isolated hearts. Coronary vascular resistance (CVR; mm Hg/mL/min/100 g) was calculated. RESULTS: The ET-1 elicited increases in CVR dose-dependently in both normotensive and hypertensive rat hearts. However, the responses were significantly greater in SHR than in WKY rat. Chronic treatment with enalapril or candesartan inhibited the development of hypertension and cardiac hypertrophy equally in SHR. Augmented vasoconstrictive responses to ET-1 were significantly reduced in treated SHR. There was no difference in these effects between enalapril and candesartan. CONCLUSION: These findings suggest that both ACE inhibitors and ARB can equally inhibit augmented coronary vascular response to ET-1 in hypertensive hearts. PMID- 11939625 TI - Covalently immobilized aminonaphthalimide as fluorescent carrier for the preparation of optical sensors. AB - By replacing the hydrogen of the 4-amino group of a 4-amino-1,8-naphthalimide derivative with an N-acryloxyethyl group, the fluorophore has been covalently immobilized on an optical sensor surface by UV photopolymerization. The optical sensor obtained can be used for the determination of picric acid. The linear range and detection limit of the sensor are 9.80x10(-7)-1.96x10(-4) mol L(-1) and 7.1x10(-7) mol L(-1), respectively. Leaching of the fluorophore from the membrane is effectively prevented by covalent immobilization, resulting in a sensor with a relatively long lifetime. The response time of the sensor is short, and the reproducibility and reversibility are good. The sensor has been used for the indirect determination of the chloroquine content of pharmaceutical tablets. PMID- 11939626 TI - Enhanced chemiluminescence for the oxidation of luminol with m chloroperoxybenzoic acid catalyzed by microperoxidase 8. AB - The use of m-chloroperoxybenzoic acid (mCPBA) in stead of hydrogen peroxide causes an increase in chemiluminescence (CL) of luminol oxidation catalyzed by microperoxidase 8 (MP8) by an order of magnitude. The accelerated formation of an intermediate plays a major role in the CL enhancement, which also leads to a significant reduction in CL duration. The presence of guanidine hydrochloride, sodium carbonate, or sodium chloride further increases the CL emission drastically. The CL emission enhancement is strongly pH dependent. The enormous enhancement of the CL signal is due to an accelerated CL cycle and an improved CL efficiency in the presence of the enhancer. The CL signal covers several orders of magnitude over a wide range of concentrations of luminol and mCPBA. The intense CL of MP8-luminol-mCPBA in the presence of the enhancer will have great potential for extremely sensitive CL assays. PMID- 11939627 TI - Determination of 41Ca in biological-shield concrete by low-energy X-ray spectrometry. AB - An X-ray spectrometric method has been developed for the determination of 41Ca in the biological-shield concrete of nuclear reactors. The concrete sample was first decomposed with nitric, hydrofluoric, and perchloric acids. Calcium was then separated from other radionuclides by ion-exchange chromatography and recovered as an oxalate precipitate. X-rays at 3.3 keV from 41Ca in the calcium oxalate pellet were measured. The detection efficiency of the X-ray measurement at 3.3 keV was calculated from those obtained by measuring 55Fe standard pellets at 5.9 keV using mass-absorption coefficients of the calcium oxalate pellet at each X ray energy. A lower limit of determination of 8 Bq g(-1) was obtained for a sample weight of 1 g. PMID- 11939628 TI - Determination of REEs distribution in monazite and xenotime minerals by ion chromatography and ICP-AES. AB - Ion chromatographic techniques were investigated for the separation and the quantitative determination of some rare earth elements (REEs) in monazite and xenotime minerals. The influences of selected eluents containing complexing acids including oxalic and alpha-hydroxy isobutyric acid (alpha-HIBA) on the retention and hence the separation efficiency of REEs was studied. Different variables affecting the separation of different REEs such as pH, type, and concentration of the mobile phase were investigated. Gradient elution, using an advanced gradient pump, was controlled automatically by the Dionex AI-450 computer software. Separation of REEs was carried out using an Ion Pac CS5A column followed by a post column derivatization reaction with 4-(2-pyridylazo)resorcinol (PAR) and UV VIS spectrophotometric detection. Mineral dissolution was carried out using sulfuric acid. A comparative evaluation of REE distribution in monazite and xenotime minerals using both ion chromatography (IC) and inductively coupled plasma atomic emission spectrometric (ICP-AES) techniques was carried out. PMID- 11939629 TI - Determination of trace phosphorus in high purity tantalum materials by inductively coupled plasma mass spectrometry subsequent to matrix separation with on-line anion exchange/coprecipitation. AB - An on-line matrix separation/inductively coupled plasma mass spectrometry (ICP MS) method is proposed for the determination of trace amounts of phosphorus in high purity tantalum metal, tantalum (V) oxide, and tantalum pentaethoxide. In the present method, the matrix tantalum in the sample solution was adsorbed on the anion exchange resin, and phosphorus (phosphate ion) was eluted with the carrier solution of HF and HNO3 mixture. Then, the effluent solution was subsequently mixed with bismuth solution and aqueous ammonia solution to coprecipitate phosphate together with bismuth hydroxide. The precipitate formed was collected on the in-line membrane filter to wash out nitric acid with pure water, and then dissolved with hydrochloric acid. The obtained phosphorus sample solution was introduced directly into the nebulizer of ICP-MS for the determination of phosphorus. Phosphorus was determined at the molecular ion signal of 31P16O+ (m/z 47). The detection limit (3sigma) of phosphorus in the present method was 1.3 ng mL(-1) as the sample solution basis, and the relative standard deviation for 30 ng mL(-1) of phosphorus in the standard solution was 4.3% in the replicate measurements (n=11). The present method was applied to the analysis of high purity tantalum materials. The concentrations of phosphorus in tantalum samples were in fairly good agreement with those obtained by glow discharge mass spectrometry (GDMS). PMID- 11939630 TI - Determination of Cr(VI) in welding fumes by anion-exchange fast protein liquid chromatography with electrothermal atomic absorption spectrometric detection. AB - The applicability of an anion-exchange fast protein liquid chromatographic electrothermal atomic absorption spectrometric procedure (FPLC-ETAAS) was investigated for the determination of Cr(VI) in welding fumes after alkaline extraction of aerosols loaded on filters. Gas tungsten arc welding (GTAW) of stainless steel was applied. Samples of welding fumes were collected during regular welding on polycarbonate membrane filters of 8 microm and 0.4 microm pore size (inhalable and respirable aerosols). Alkaline extraction (2% NaOH-3% Na2CO3) of filters in a heated ultrasonic bath was applied to leach Cr from the airborne particulate matter. 0.5 cm3 of sample extract was then injected onto an anion exchange FPLC column. Tris-HCl buffer (0.005 mol dm(-3), pH 8.0) and the same buffer with NaCl (0.5 mol dm(-3)) were employed in gradient elution (15 min, flow rate 1 cm3 min(-1)). The separated Cr species were determined "off line" by ETAAS in 0.5 cm3 fractions. Cr(VI) was reproducibly and quantitatively eluted from 12.0 to 13.0 min with a maximum peak at 12.5 min. Good repeatability of measurement (+/-3.0%) of alkaline extracts was obtained for Cr(VI). The LOD (3s) was found to be 0.035 microg m(-3) Cr(VI), when 2 m3 of aerosols were collected on the filter. Validation of the procedure was performed by spiking alkaline extracts and by the analysis of standard reference material CRM 545, Cr(VI) in welding dust loaded on a filter. The technique was successfully applied for the determination of Cr(VI) in welding fumes. PMID- 11939631 TI - Determination of alkylphenols and alkylphenol polyethoxylates by reversed-phase high-performance liquid chromatography and solid-phase extraction. AB - A simple, accurate and reproducible reversed-phase high-performance liquid chromatography (HPLC) method was developed for the separation and characterisation of alkylphenols (APs) and alkylphenol polyethoxylates (APEOs), using a C18 octadecyl silica (ODS) column. APs and each APEO oligomer were separated successfully within a reasonable time without gradient elution. An excellent resolution was obtained, even for mixtures of APs and low EO number APEOs, which are otherwise difficult to separate using conventional normal-phase HPLC methods. This method, combined with solid-phase extraction, was highly applicable for the simultaneous determination of alkylphenols and alkylphenol ethoxylates in real samples. PMID- 11939632 TI - Comparison of different extraction techniques for the determination of polychlorinated organic compounds in sediment. AB - The performance of various enhanced extraction techniques, such as accelerated solvent extraction, microwave-assisted extraction, Soxhlet extraction, ultrasonic extraction and fluidized-bed extraction for the determination of polychlorinated organic compounds in dry sediment was investigated in two laboratories. The results of the two laboratories were in good agreement. The extraction yields from the batch extraction methods were lower than those from the dynamic techniques. Accelerated solvent extraction, especially, exhibited higher extraction efficiency than the standard procedure, Soxhlet extraction, whereas the results of fluidized-bed extraction were comparable. PMID- 11939633 TI - Metabolites from the biodegradation of pharmaceutical residues of ibuprofen in biofilm reactors and batch experiments. AB - The three metabolites hydroxyibuprofen (OH-Ibu), carboxyibuprofen (CA-Ibu), and carboxyhydratropic acid (CA-HA), also known from human metabolism of ibuprofen, could be identified in biodegradation experiments. Identification was based on EI mass spectra and comparison with literature data. Detection was performed by selective MS-MS measurements by GC-ion-trap MS and online methylation. Ibuprofen (Ibu), OH-Ibu, and CA-Ibu could be detected with a signal-to-noise ratio of 10:1 at a concentration of 2 nmol L(-1), CA-HA at 0.5 nmol L(-1). Degradation experiments in both biofilm reactors (BFR) and batch experiments with activated sludge (BAS) reveal OH-Ibu as the major metabolite under oxic conditions, and CA HA under anoxic conditions. CA-Ibu was found under oxic and anoxic conditions almost only in the BAS. The metabolites together do not account for more than 10% of the initial concentration of Ibu. PMID- 11939634 TI - Determination of selenium in blood serum by inductively coupled plasma atomic emission spectrometry with pneumatic nebulization. AB - The possibility of determining selenium in blood serum using inductively coupled plasma emission spectrometry with conventional pneumatic nebulization was studied. A high-resolution spectrometer (SBW=6 pm) with laterally viewed ICP was employed. Analysis with conventional pneumatic nebulization could overcome laborious and demanding digestion, which is necessary for hydride generation. A pressure digestion with nitric acid at 160 degrees C was sufficient to decrease the carbon content in the serum sample to 5%-10% of its original value. Spectral interference of the CN band was observed and mathematically corrected. It was found that the carbon-induced selenium line emission enhancement occurred even under ICP optimized conditions. A method of determination was developed and applied to the analysis of blood serum. True limit of detection in real samples is 0.01-0.02 mg/L and the limit of quantification (RSD 10%) is 0.03-0.07 mg/L using Se I 196.090 nm line at an integration time of 10-2 s. The method was tested by analysis of porcine blood serum and the serum reference material Seronorm MI 0181. PMID- 11939635 TI - Electrochemical study of fluvastatin sodium--analytical application to pharmaceutical dosage forms, human serum, and simulated gastric juice. AB - The oxidation of fluvastatin sodium on a glassy carbon electrode has been studied by use of a variety of voltammetric techniques. Different conditions were investigated to optimize the determination of fluvastatin sodium. The dependence of the intensities of currents and potentials on pH, concentration, scan rate, and nature of the buffer was investigated. Oxidation of fluvastatin sodium was found to be diffusion-controlled and irreversible. The best results for the determination of fluvastatin sodium were obtained by using differential pulse and square-wave voltammetric techniques in Britton-Robinson buffer at pH 10.04. Differential pulse and square-wave voltammetry at a glassy carbon electrode resulted in linear calibration in the range 8x10(-6) to 6x10(-4) mol L(-1) and detection limits of 1.07x10(-6) and 7.99x10(-7) mol L(-1), respectively. The proposed methods were successfully applied to the determination of the drug in capsules and biological fluids. Excipients did not interfere with the determination. Statistical validation revealed that the methods were free from significant systematic errors. PMID- 11939636 TI - A manifold for the automatic dilution of concentrated solutions in flame atomic absorption spectrometry. AB - A fully computer-controlled manifold is described which enables the automatic dilution and measurement of solutions which are too concentrated to be measured by direct aspiration in FAAS. The sample solution is propelled by a peristaltic pump equipped with two pump tubes of different diameters, the difference between the nebulizer uptake rate and the flow rate supplied by both tubes being compensated by solvent provided through a T-piece. The absorbance is averaged and the pump turning speed automatically increased or reduced until an absorbance value within the linear response of the spectrometer is obtained. To furnish very high dilution, the system automatically reverses the direction of the flow moved through one of the pump tubes, so that the net flow delivered by the pump is the difference between the flows propelled through the two pump tubes. In these circumstances, in addition to absorbance, the module of the Fourier transform obtained from the absorbance-time profile can be used as the analytical signal. Calibration is performed by use of a single standard solution. In this way, it is possible to determine copper in the 1-10,000 microg mL(-1) range with an RSD of 1.3-3%. Copper, zinc, calcium, and magnesium were determined in different samples to check the reliability of the procedure. PMID- 11939637 TI - Hazards of errors in the palladium storage in determination by GFAAS. AB - Losses of palladium in weakly acidic solutions during storage process, which occur by its hydrolysis and/or adsorption onto the surface of storage vessels, are studied and ways of overcoming them are proposed. A fast and effective procedure for cleaning of laboratory ware with 0.2 mol L(-1) thiourea in 0.1 mol L(-1) HCl is described. Preparing palladium solutions in quartz vessels as well as using autosampler cups made from quartz glass, enabled to diminish the detection limit of palladium in synthetic solutions by GFAAS from 0.94 ng/mL to 0.27 ng/mL. PMID- 11939638 TI - ICP-AES determination of traces of noble metal ions pre-concentrated and separated on a new polyacrylacylaminothiourea chelating fiber. AB - A new polyacrylacylaminothiourea chelating fiber was synthesized simply and rapidly from nitrilon (an acrylonitrile-based synthetic fiber) and aminothiourea. This fiber was used for the pre-concentration and separation of traces of AuIII, PtIV, PdIV, and IrIV ions from aqueous samples. Factors effecting and parameters for the adsorption of these ions onto the resin, such as acidity, rate, reuse, capacity, and interference of other ions, were investigated using inductively coupled plasma atomic emission spectrometry (ICP-AES). Conditions of desorption of these ions from the resin were similarly investigated. The enrichment factor was 100. The relative standard deviation for the determination of AuIII, PtIV, PdIV, and IrIV were in the range of 0.7-3.0% for 20.0 ng/mL of each. The recoveries of a standard from real solution samples were between 96% and 100%. The concentrations of analyzed ions in a digested metal powder sample processed with the method proposed were in good agreement with the certified values. PMID- 11939639 TI - A novel chemiluminescent method for the determination of salicylic acid in bactericidal solutions. AB - A new flow-injection procedure has been developed for the determination of salicylic acid based on the enhancement of the chemiluminescence from the cerium(IV)-Tween 20 reaction by salicylic acid in acidic medium. The method is simple, selective and sensitive with a detection limit of 2.5x10(-9) g mL(-1). It is applicable to the determination of salicylic acid in the concentration range of 4.0x10(-9)-1.1x10(-6) g mL(-1). The relative standard deviation (RSD) is 0.85% for 4.0x10(-7) g mL(-1) salicylic acid (n=11). The method has been successfully applied to the determination of salicylic acid in bactericidal solutions. Furthermore, it is suggested that light emission from cerium(IV)-Tween 20 reaction is probably because of the formation of singlet oxygen 1O2* and the emitter is excited oxygen molecular pairs O2(1delta(g))O2(1sigma(g)-). PMID- 11939640 TI - Inhaled nitric oxide in infants and children. AB - NO has been used successfully to treat PPHN, reducing the need for ECMO. NO has also been used in the cardiac catheterization laboratory to determine if pulmonary hypertension will decrease with NO. Patients who do not respond to NO are at higher risk after open-heart surgery, because their pulmonary hypertension will be difficult to treat. Postoperatively, NO can be used to determine if pulmonary hypertension is caused by vasoconstriction or by an obstruction. Inhaled Nitric Oxide at a Glance: ACTION: Selective pulmonary vasodilation without systemic vasodilation. USE: Treatment of pulmonary hypertension. CONCENTRATION AND ROUTE: Lowest concentration that will produce pulmonary vasodilation and improved oxygenation. Concentration should be kept < 80 ppm. CONTRAINDICATION: Neonate that is ductal-dependent. TOXIC EFFECTS: Keep methemoglobin level < 5%. Keep nitric dioxide, which can cause lung damage, < 7 ppm. Risk of bleeding. MONITOR: Levels of NO/NO2. Platelets. Arterial blood gas (ABG). Methemoglobin. WEANING: Decrease NO by 20%, monitoring ABG at 3- to 4-hour intervals. If there is a decrease in oxygenation, increase NO. Increase FIO2 20% when NO is discontinued. Unsuccessful treatment with NO--keep on NO until ECMO is available. PMID- 11939641 TI - Medications and the elderly in the critical care setting. AB - As the elderly population in our country continues to increase, it is imperative that the bedside practitioner be keenly aware of the factors that affect pharmacokinetics and pharmacodynamics of medication administration. Refining the admission interview process to obtain accurate medication information--including alternative and OTC drugs--will help to decrease the incidence of drug interactions and ADR in the acute care setting. Appropriate monitoring of pharmacologic effects and organ function will allow the practitioner to better treat the elderly client with as few medications as possible. PMID- 11939642 TI - Treatment of gram-positive infections: past, present, and future. AB - The need for strategic planning for antimicrobial use has reached a critical point. The rise in resistant nosocomial and community gram-positive bacteria mandates appropriate antibiotic selection and dosing. The development of new compounds is not the answer, because many are based off existing structures to which bacteria have already developed resistance. New antimicrobial agents are falling to the resistant mechanisms developed by the bacteria, after only limited clinical exposure. Judicious use of antimicrobial agents and applying pharmacokinetic principles when dosing can help slow the rate of resistance. PMID- 11939643 TI - Pharmacology advances in the neuroscience intensive care unit. AB - New drugs are continuously being developed to enhance care in the neuro ICU. Newer drugs with special implications for the neurologically impaired population include fosphenytoin, tissue plasminogen activator, and propofol. Nurses are challenged to use these drugs in a safe and effective manner. PMID- 11939644 TI - Pharmacologic prophylaxis and treatment of stress ulcers in critically ill patients. AB - Most clinicians agree that critically ill patients are at significant risk of developing stress-related ulcers and may have already developed mucosal lesions even if they are asymptomatic. Many options, including new pharmacologic advances, are available for the treatment and prophylaxis of stress-related ulcers; therefore, all critically ill patients should receive prophylaxis, even if they do not require treatment. Nutrition may play a significant role in the future in preventing stress-related ulcers. By improving stores of critical elements such as antioxidants, vitamins, and minerals before surgery, patients may lower the risk of developing stress ulcers. Critical care clinicians are critical in preventing and treating stress-related ulcers and should be vigilant in their patient care. PMID- 11939645 TI - Antidysrhythmic agents at the turn of the twenty-first century: a current review. AB - The use of class IA agents is gradually on the decline, primarily as a result of its unfavorable risk-to-benefit ratio. Lidocaine, a class IB agent, has been widely used in the acute treatment of VT. However, alternate drugs are being considered increasingly as first-line agents in the acute treatment of VT. Class IC drugs are contraindicated in patients with structural cardiac abnormalities and have a limited usefulness in the management of dysrhythmias. Beta-blockers continue to increase their role in cardiology, and subsequently their use in managing dysrhythmias. Class III agents, including amiodarone, sotalol, ibutilide, and dofetilide, are among the most widely used antidysrhythmics. Class IV calcium channel blockers have a limited usefulness in tachydysrhythmias. Digoxin and adenosine have unique antidysrhythmic properties and will likely retain their roles as antidysrhythmic agents. In the wake of the effectiveness of amiodarone, the drug that crosses all classes, some now question the benefit of pure agents that block a single, specific ion channel in the heart. After CAST8 demonstrated that antidysrhythmics can increase mortality while seemingly suppressing dysrhythmias, new drugs will continue to undergo intense scrutiny with regard to their efficacy, safety, and usefulness in treating dysrhythmias. PMID- 11939646 TI - Drugs used to limit blood surface interactions. AB - Since the days of the triumvirate of aspirin, heparin, and warfarin, health care practitioners have attempted to blunt the response of the coagulation system. Use of CPB has enhanced our awareness of the effects that a foreign surface has on blood in general. Success in correcting or preventing life-threatening conditions caused by thromboembolic disease depends on having the ability to correct the condition as well as the ability to limit the body's response to the intervention. Knowing how the body responds to foreign surfaces is crucial, as is awareness of current methods to limit these responses. This approach will help ensure procedural success with fewer side effects and better overall patient outcomes. PMID- 11939647 TI - Critical care hemodynamic parameters and pharmacologic interventions. AB - Hemodynamics are a critical component in treatment of serious illness in the ICU setting. Understanding hemodynamic parameters and their implications for care is a cornerstone of critical care nursing. Appropriate interventions for hemodynamic instability and a thorough understanding of the rationale for the intervention make the difference for the critically ill patient. PMID- 11939648 TI - Drug use in renal failure. AB - A number of medications used in the presence of renal disease require dose adjustments be made. The health care professional should know a drug's route of elimination, the percentage of protein binding, and the percentage of a normal dose removed by dialysis. This information is a guideline and should be used to start therapy. The clinician must understand that individual adjustments are the rule rather than the exception. PMID- 11939649 TI - Pharmacologic treatment of the critically ill patient with diabetes. AB - The pharmacologic approach to the management of DKA and HHNS involves the use of insulin, fluids, and electrolytes. Essential to the successful treatment of DKA and HHNS is a carefully planned post-intensive-care transition period. This period is based on the understanding that insulin therapy is never interrupted in patients with type 1 diabetes and the transition to insulin and/or oral hypoglycemic therapy in type 2 patients is strategically planned. The use of acute adjuvant insulin-glucose therapy in acute coronary syndromes and cardiac surgeries can result in decrease in overall morbidity and mortality in patients with diabetes. Ultimately, however, the major approaches to the treatment of diabetes is primary prevention of diabetes itself and the secondary prevention of complications through vigorous glucose control. PMID- 11939650 TI - The next generation of medications for kidney transplant patients. AB - Transplant pharmacotherapy evolves as new agents are investigated and approved for use. Clinical immunosuppression has been plagued with maintaining a balance between rejection of the transplanted organ and complications of over immunosuppression, including infection and malignancy. Clinicians must understand current immunosuppressive regimens and their associated effects when caring for transplant patients. While all transplant patients receive some form of immunosuppressive therapy, the combinations and choices increase as new drugs are developed. In the critical and acute care settings, newly transplanted patients will likely receive induction therapy. The goal of induction therapy is to increase long-term patient and allograft survival while preventing or reducing rejection episodes. Several agents are available for induction therapy, and each transplant center designs its own protocol. The foundation for maintenance therapy rests on the combining immunosuppressives to prevent rejection through a variety of pathways. An understanding of the mechanism of action and additive effects of a drug allows practitioners to optimize therapy while decreasing adverse effects. Immunosuppressive therapy offers potential for reducing detrimental patient outcomes and improving allograft survival. It is well established that repeated rejection episodes correlate with poor long-term graft survival. Challenges facing researchers and clinicians focus on improved patient outcomes and options to address financial constraints of transplantation. PMID- 11939651 TI - Surgery in the elderly. AB - The elderly (those 75 years of age or older) are a heterogeneous group. They present with both elective and urgent surgical problems, and risk assessment, decision-making and perioperative care are typically more challenging than in younger patients. An appreciation for this heterogeneity and an understanding of how physiologic changes of aging affect surgical care are essential if the best outcomes are to be achieved. PMID- 11939652 TI - Users' guide to the surgical literature: how to use an article reporting population-based volume-outcome relationships in surgery. PMID- 11939653 TI - Functional outcome of total knee arthroplasty after high tibial osteotomy. AB - OBJECTIVE: To evaluate the functional outcome for patients who undergo total knee arthroplasty (TKA) after high tibial osteotomy (HTO). DESIGN: Retrospective matched cohort study. SETTING: University of Toronto affiliated hospital. PATIENTS: Twenty patients who underwent TKA after HTO and 20 matched patients who received a primary TKA. INTERVENTION: TKA. OUTCOME MEASURES: The Medical Outcomes Study Short Form (SF-36) health survey score and the Western Ontario and McMaster Universities (WOMAC) Osteoarthritis Index. Univariate analyses were used to compare the case and control groups with respect to baseline variables using the t-test, chi2 test or Fisher's exact test. Functional outcomes were assessed by multivariate analyses. RESULTS: Operative problems were more frequently encountered in the study group, which had longer operative times (p < 0.0001), more difficulties with patellar eversion (p = 0.021) and an increased number of lateral releases performed (p = 0.0089). There were trends toward a significant difference in the pain (p = 0.07), function (p = 0.18) and stiffness (p = 0.14) categories of the WOMAC Osteoarthritis Index between the 2 groups, suggesting poorer functional outcomes of TKA after HTO, but the results did not reach statistical significance. A previous HTO does not affect the general health of patients after TKA, as there was no difference between the 2 groups in SF-36 scores. CONCLUSIONS: TKA after HTO is a technically more challenging procedure than primary TKA. The functional outcomes at a mean follow-up of 5 years after TKA in patients with a previous HTO tended to be inferior but the differences were not significant (p > or = 0.05). PMID- 11939654 TI - Presidential address, 2001. Advice to young surgeons. AB - In his 2001 presidential address to the Canadian Association of General Surgeons, the author offers advice to young surgeons, based on his lifetime experience as a surgical educator, researcher and practitioner. He offers the following samples of wisdom for young surgeons: they should be prepared for a lifetime of learning and be willing and able to adapt to new advances; they should listen to their patients as they describe their presenting complaints and not be tempted to interrupt; they should take time in an emergency situation and remember that split-second decisions can affect the patient for a lifetime; they should be willing to take advice from fellow professionals; they should take time to maintain a quality family life and take adequate time away from the workplace; they should be active be a role model in their community; and, finally, they should get involved and adopt an advocacy role in their profession. PMID- 11939655 TI - Presidential address, 2001. Roots and relevance. AB - The infrastructure, focus and modus operandi of the Canadian Orthopaedic Association (COA) has been changed to effectively address the needs of the Association in this new century, as well as the possibility of a diminished interest in orthopedic surgery as a career choice. These issues are dealt with in this COA presidential address given at the COA annual general meeting in June 2001. PMID- 11939656 TI - Multiple trauma in children: predicting outcome and long-term results. AB - OBJECTIVE: To analyze the management of pediatric trauma and the efficacy of the Pediatric Trauma Score (PTS) in classifying injury severity and predicting prognosis. DESIGN: A retrospective case series. SETTING: The Children's Hospital of Eastern Ontario, a major pediatric trauma centre. PATIENTS: One hundred and forty-nine traumatized children with 2 or more injuries to 1 body system or a single injury to 2 or more body systems. INTERVENTIONS: Use of the PTS and Glasgow Coma Scale score in trauma management. MAIN OUTCOME MEASURES: Types of injuries sustained, complications, missed injuries, psychosocial effects and residual deficiencies. RESULTS: The average PTS was 8.5 (range from -3 to 11). The total number of injuries sustained was 494, most commonly closed head injury (86). Forty-two percent of children with an average trauma score of 8.5 were treated surgically. There were 13 missed injuries, and complications were encountered in 57 children, the most common being secondary to fractures. Forty eight (32%) children had residual long-term deficiency, most commonly neurologic deficiency secondary to head injury. CONCLUSIONS: Fractures should be stabilized early to decrease long-term complications. A deficiency of the PTS is the weighting of open fractures of a minor bone. For example, metacarpal fracture is given the same weight as an open fracture of the femur. Neuropsychologic difficulties secondary to trauma are a major sequela of trauma in children. PMID- 11939658 TI - Cellular schwannoma: a rare presacral tumour. PMID- 11939657 TI - Factors influencing the adoption of blood alternatives to minimize allogeneic transfusion: the perspective of eight Ontario hospitals. AB - OBJECTIVE: To identify and describe the factors influencing the use and nonuse of blood-sparing methods such as preoperative autologous donation, acute normovolemic hemodilution, and the use of cell salvage devices, hemostatic agents and erythropoietin. DESIGN: An interview survey. SETTING: Eight Ontario hospitals. METHOD: Interviews were conducted with chiefs of surgery, orthopedics, cardiac surgery and anesthesia, and with heads of transfusion medicine and pharmacy. Hospitals were selected using the qualitative sampling strategy of maximum variation based on their use of the methods (as reported in a previous mail survey). RESULTS: Use of blood-sparing methods was influenced by diverse factors often operating simultaneously. These included the following: characteristics of the method (e.g., evidence of its effectiveness, ease of use, cost); perceptions and experiences of the potential adopters (experience with the method, perception of the current safety of allogeneic blood, perceived convenience or inconvenience of using the method); aspects of the practice setting (inability to move resources between hospital departments, presence of a local clinical champion); and the external environment (patient and public expectations, funding of the blood system, blood shortages). INTERPRETATION: More rational and evidence-based use of blood-sparing methods could be promoted by the adoption of an interdisciplinary, comprehensive, coordinated approach tailored to each patient's needs. PMID- 11939659 TI - Intrathoracic hibernoma. PMID- 11939660 TI - An early 19th-century Canadian surgical practice: the casebook of John Mackieson of Charlottetown, 1795-1885. AB - A casebook written by Dr. John Mackieson (1795-1885), of Charlottetown, contains the records of 49 surgical cases he managed between 1826 and 1857. In view of the rarity of first-hand accounts of surgical practice in Canada in the mid-19th century, Mackieson's case records are a significant source of information. These cases are discussed in order to delineate Mackieson's approach to the surgical problems he faced in his general practice. His case records also illustrate some of the general problems that beset surgeons in that era. PMID- 11939661 TI - Authorship. PMID- 11939662 TI - The future of rural surgical care in Canada: a time for action. PMID- 11939663 TI - Soft-tissue images. Pseudomyxoma peritonei and villous adenoma of the appendix. PMID- 11939664 TI - Musculoskeletal case 21. Insufficiency fracture of the sacrum. PMID- 11939665 TI - Soft-tissue case 44. Recurrent epiploic appendagitis. PMID- 11939666 TI - SESAP questions. Category 6, items 43-44. Treatment priorities in combined thoracic aortic injury and intra abdominal injury. PMID- 11939667 TI - An expanding role for cardiopulmonary bypass in trauma. AB - OBJECTIVES: To analyze experience at the McGill University Health Centre with cardiopulmonary bypass (CPB) in trauma, complemented by a review of the literature to define its role globally and outline indications for its expanded use in trauma management. DATA SOURCES: All available published English-language articles from peer reviewed journals, located using the MEDLINE database. Chapters from relevant, current textbooks were also utilized. STUDY SELECTION: Nine relevant case reports, original articles or reviews pertaining to the use of CPB in trauma. DATA EXTRACTION: Original data as well as authors' opinions pertinent to the application of CPB to trauma were extracted, incorporated and appropriately referenced in our review. DATA SYNTHESIS: Overall mortality in the selected series of CPB used in the trauma setting was 44.4%. Four of 5 survivors had CPB instituted early (first procedure in operative management) whereas 3 of 4 deaths involved late institution of CPB. CONCLUSIONS: Although CPB has traditionally been used in the setting of cardiac trauma alone, a better understanding of its potential benefit in noncardiac injuries will likely make for improved outcomes in the increasingly diverse number of severely injured patients seen in trauma centres today. Further studies by other trauma centres will allow for standardized indications for the use of CPB in trauma. PMID- 11939668 TI - Antihypertensive effect of stevioside in different strains of hypertensive rats. AB - BACKGROUND: Stevioside is a natural sweet-tasting glycoside isolated from the herb Stevia rebaudiana, composed of stevia, a diterpenic carboxylic alcohol with three glucose molecules, mainly used commercially as sugar substitute. Previous study has shown that it can lower blood pressure in anesthetized spontaneously hypertensive rats (SHR). This study was undertaken to evaluate the antihypertensive effect of stevioside in different strains of hypertensive rats and to observe whether there is difference in blood pressure lowering effect. METHODS: Noninvasive tail-cuff method was employed to measure blood pressure. Stevioside at the concentrations of 50, 100 and 200 mg/kg were administered intraperitoneally (ip) to normotensive Wistar-Kyoto rats (NTR), SHR, deoxycorticosterone acetate-salt (DOCA-NaCl) sensitive hypertensive rats (DHR) and renal hypertensive rats (RHR). RESULTS: Significant hypotensive effect of stevioside administered ip was noted in different strains of rats at the dose of 50 mg/kg. When stevioside was increased to the concentrations of 100 and 200 mg/kg, ip, it also caused slow and persistent lowering of blood pressure in SHR and NTR. Data also showed that stevioside given at the concentrations of 100, 200 and 400 mg/kg ip resulted in lowering of blood pressure in SHR dose-dependently. Blood pressure returned to previous levels after the drug was discontinued for 2 3 days. Drinking of 0.1% stevioside solution in mature SHR could have antihypertensive effect and also prevented hypertension in immature SHR. CONCLUSIONS: This study reconfirmed stevioside has hypotensive effect and the effect is more prominent in hypertensive rats. PMID- 11939669 TI - Clinical pathway: impact on the future work of all physicians? PMID- 11939670 TI - Pulmonary resection for metastases from colorectal adenocarcinomas. AB - BACKGROUND: Colorectal cancer is one of the most common cancers in Taiwan, as in the other part of the world. Surgical intervention is the best treatment of choice, yet some patients still developed distant metastasis after primary lesions were controlled. Metastasectomy is reported to improve the survival. This retrospective study was carried out to evaluate the relationship between the prognostic factors of lung metastasectomy and patient survival. METHODS: From 1981 to 2000, 68 patients undergoing complete lung metastasectomy in our section were studied. The prognostic factors influencing survival were analyzed, and the survival analysis was made with Kaplan-Meier method and compared by log-rank test. RESULTS: There was no surgical mortality in our series. The 5-year survival was 36.1%. None of parameters such as age, sex, stage of primary colorectal cancer, surgical method, size, number of metastatic deposits and disease free interval showed to relate with the survival. CONCLUSIONS: We concluded that lung metastasectomy for colorectal cancer is safe and effective to improve the survival. Since there were no significant factors influencing the survival, there should be no absolute contraindication against resectable pulmonary metastasis after primary lesions were adequately controlled. PMID- 11939671 TI - Effects of period-free hormone replacement therapy in postmenopausal women in Taiwan. AB - BACKGROUND: To compare the effects of continuous combined hormone replacement therapy (Kliogest) and placebo on climacteric symptoms, bone turnover markers, serum lipid profiles and the safety of Kliogest. METHODS: This is a 4-month, single-centre, double-blind, placebo-controlled clinical study. Fifty-six healthy volunteer women with intact uterus, between six months and three years after a natural menopause, were recruited and randomised into two groups to have blood and urine laboratory test for assessment of bone resorption marker of deoxypyridinoline, bone formation marker of bone specific alkaline phosphatase, lipid profile including total cholesterol, high-density lipoprotein, low-density lipoprotein and triglyceride, and menopausal symptoms. Information was taken at initial visit and 1, 2 and final follow-up after taking medication. RESULTS: Continuous Kliogest treatment resulted in significant reduction in menopausal symptoms, as scored by Integrated Greene Climacteric Scale and in the psychological, somatic and vasomotor subscales scores (p = 0.009). Kliogest reversed the negative trends in lipoprotein profiles by lowering total cholesterol (p < 0.001), low-density lipoprotein cholesterol (p < 0.001), and triglyceride (p = 0.004), but decreasing high-density lipoprotein cholesterol (p = 0.002). A significant reduction in bone resorption marker, deoxypyridinoline, was also observed in the Kliogest group (p < 0.001). There was no report of serious adverse events, endometrial hyperplasia or cancer in the Kliogest treated patients. CONCLUSIONS: Four-month treatment with Kliogest resulted in an improvement in menopausal symptoms and lipid profiles as well as reduction in bone loss. PMID- 11939672 TI - Is combined abdominal aortic aneurysm repair and coronary artery bypass grafting feasible? AB - BACKGROUND: Significant coronary artery disease requiring coronary artery bypass grafting (CABG) may co-exist with large abdominal aortic aneurysm (AAA) in some patients. We reviewed our experience in either staged or simultaneous operation. METHODS: The records of all patients receiving both CABG and AAA repairs in recent 7 years were retrospectively reviewed. The patient demographics, severity of coronary disease, AAA size, duration of staged procedures, perioperative morbidity and mortality rates as well as the hospital cost were analyzed. RESULTS: From June 1993 to Sept 2000, totally 14 patients received both CABG and AAA repair, including 6 patients for simultaneous operation (group A, 42.8%) and 8 for staged operation (group B, 57.2%) with CABG first. Patients in the group A were younger and with larger AAA. There was neither operative mortality in both group nor interprocedure AAA rupture in group B. Total postoperative hospital stay and hospital cost were significantly decreased in group A than in group B rehospitalized patients. CONCLUSIONS: Simultaneous CABG and AAA repair is feasible in surgical technique. In those younger patients with larger AAA, combined surgery could be performed as safely as staged procedures. PMID- 11939673 TI - Management of laryngeal amyloidosis--the experience of Taipei Veterans General Hospital. AB - Laryngeal amyloidosis is an uncommon disease accounting for 0.68% of benign tumors of the larynx. It can present as a nodular tumor or diffuse subepithelial deposition forming a bulging mass and causing hoarseness, dysphagia or even airway obstruction. We present our experience in the management of laryngeal amyloidosis, and discuss the clinical features and histopathological characteristics of the disease. Reports on benign laryngeal tumor from June 1988 to June 1998 at Taipei VGH were searched to identify and collect amyloid cases for chart review. Five cases of laryngeal amyloidosis were found in the records. The sites of involvement were false cord, ventricle, post-cricoid, and arytenoid epiglottic fold. No evidence of systemic amyloidosis involvement was observed for any of the five cases. All cases were treated with CO2 laser excision. Recurrence occurred in two patients within one year. More extensive CO2 laser excision was subsequently performed, with no more recurrences noted as of the last follow-ups. Laryngeal amyloidosis is a benign disease; however, surgical management is required when it is symptomatic. Recurrences or large-sized tumors may be a problem; however, advances in laser technology and the cumulative experience of treating benign lesions of the larynx have made CO2 laser endoscopic excision an effective treatment option. PMID- 11939674 TI - Laryngeal amyloidosis: long term follow-up is needed due to its potential for recurrence. PMID- 11939675 TI - Multifocal retinoblastomas treated with transpupillary thermotherapy. PMID- 11939676 TI - Boerhaave syndrome with atypical clinical presentations diagnosed by computed tomography. AB - Spontaneous rupture of esophagus (Boerhaave syndrome) is a life-threatening emergency requiring immediate surgical management. The diagnosis is often delayed and results in high mortality and morbidity. Typical clinical presentations are postprandial vomiting and subsequent chest or back pain, which alert the clinicians to take chest film and esophagogram for diagnosis. We presented a case of Boerhaave syndrome with atypical presentation mimicking dissecting aortic aneurysm, which was diagnosed by computed tomography. PMID- 11939677 TI - Effect of a clinical pathway on selected clinical outcomes of pulmonary lobectomy. AB - BACKGROUND: North American hospitals use clinical care pathways to reduce length of stay, readmissions, and resource utilization and also to increase patient satisfaction. This study examined the effects of clinical pathways after pulmonary lobectomy in Taiwanese patients. METHODS: During 1997, a multidisciplinary team developed a lobectomy clinical pathway. The Program Evaluation Review Technique was used to analyze variances from the clinical pathway. Based on these findings, a standardized clinical pathway was implemented in late 1997. Forty patients participated. The variables of the study are length of hospital stay, readmission rates, spirometry usage, patient education and costs benefited from clinical care pathway use. RESULTS: Fourteen lobectomy patients following the clinical pathway had a mean length of stay of 17.9+/-4.18 days (p < 0.001) and 0% readmission (p < 0.001). Without a pathway, 26 lobectomy patients had a mean length of stay of 37.5+/-6.18 days and 18% were readmitted. Factors affecting clinical pathway success were preoperative days(p < 0.001), postoperative days (p = 0.033), spirometry usage (p = 0.043) and patient education (p = 0.02). Clinical pathway use reduced mean hospital costs by 16% for lobectomy. CONCLUSIONS: Length of hospital stay, readmission rates, spirometry usage, patient education and costs benefitted from clinical care pathway use. Factors critical to success appear to be multidisciplinary teamwork and communication. PMID- 11939678 TI - Clinical monitoring of direct thrombin inhibitors using the ecarin clotting time. PMID- 11939679 TI - National Pharmacy Cardiovascular Council treatment guidelines for the management of type 2 diabetes mellitus: toward better patient outcomes and new roles for pharmacists. PMID- 11939680 TI - Changes in pharmacy practice faculty 1995-2001: implications for junior faculty development. AB - OBJECTIVE: To compare changes in United States pharmacy practice faculty demographics from 1995-2001 and to discuss the implications for junior faculty development. METHODS: Demographic data were extracted from the American Association of Colleges of Pharmacy institutional research system for academic years 1995-1996 and 2000-2001. RESULTS: In 2000-2001, pharmacy practice was the largest faculty discipline, 3.8 times larger than the next three disciplines. Junior pharmacy practice faculty occupied 65% of all junior full-time pharmacy faculty positions. Tenure track assistant professors decreased 4% from 283 to 271, and nontenure track assistant professors increased 58% from 427 to 677 (chi2 = 20.0, p<0.05). In 2000-2001, 72% of all pharmacy practice assistant professors were nontenure track, up from 59% in 1995-1996. Women assistant professors in pharmacy practice outnumbered men by 2:1. Challenges faced by new faculty include balancing teaching, practice, and research demands; selecting a nontenure or tenure track and understanding its expectations; limiting teaching preparation time; developing productive writing habits; setting performance goals; managing time; and handling the mental and physical stress of academic life. Senior faculty must actively help new members appreciate the many positive aspects of academic life by sharing their strategies and success stories. CONCLUSION: Schools and colleges of pharmacy relied heavily on increasing the number of nontenure track junior pharmacy practice faculty to meet increased clinical education demands. PMID- 11939681 TI - Evaluation of survival and pharmacodynamic relationships for five fluoroquinolones in a neutropenic murine model of pneumococcal lung infection. AB - STUDY OBJECTIVE: To compare the antistreptococcal activity of five fluoroquinolone antibiotics, using a neutropenic murine model of pneumococcal pulmonary infection. DESIGN: Animal experiment. SETTING: University-affiliated research center. ANIMALS: Neutropenic and control mice weighing 24-29 g. INTERVENTION: After induction of neutropenia, renal failure, and infection with Streptococcus pneumoniae, the mice received one of five fluoroquinolones twice/day for 72 hours beginning 12 hours after infection. Dosages were selected to approximate 0.1 x AUC0-24 (area under the concentration-time curve from 0-24 hours) and AUC0-24 achieved in humans. Control mice received normal saline. Survival was assessed at regular intervals for up to 10 days. At least 10 mice were included in each cohort (range 10-34). MEASUREMENTS AND MAIN RESULTS: Ciprofloxacin, clinafloxacin, grepafloxacin, levofloxacin, and moxifloxacin were studied at subtherapeutic and therapeutic dosages against three quinolone susceptible isolates of S. pneumoniae that lacked mutations in parC, parE, and gyrA. Pharmacokinetic profile of each agent and dosing regimen was determined. A composite survival curve for all fluoroquinolones and isolates was constructed. Relationships between survival rate at 72 hours and AUC:MIC (minimum inhibitory concentration), peak:MIC, time above the MIC (percentage of dosing interval) for total and free drug concentrations were fit by using a sigmoid maximal effect (Emax) model. Survival was significantly better in the higher dosage group than in the lower dosage group. Time above MIC did not display a correlation with outcome. The AUC:MIC showed a greater correlation with outcome (R2 = 0.56 total, 0.54 free) than did peak:MIC (R2 = 0.52 total, 0.51 free). With use of composite data, total AUC:MIC ratios associated with 50%, 90%, and 99% of Emax were 34:1, 56:1, and 95:1, respectively CONCLUSIONS: In this model, efficacy was achieved with the fluoroquinolone antibiotics at dosages yielding AUC0-24 comparable to those obtained in humans. One pharmacodynamic parameter (i.e., AUC:MIC) may be applied to various fluoroquinolones and isolates of S. pneumoniae. The AUC:MIC was more predictive of outcome than was time above the MIC or peak:MIC. PMID- 11939682 TI - Clinical efficacy and pharmacoeconomics of a continuous-infusion piperacillin tazobactam program in a large community teaching hospital. AB - STUDY OBJECTIVE: To compare continuous versus intermittent administration of piperacillin-tazobactam with regard to clinical, microbiologic, and economic outcomes. DESIGN: Prospective, open-label controlled study SETTING: Community teaching hospital. PATIENTS: Ninety-eight hospitalized patients prescribed piperacillin-tazobactam. INTERVENTION: Substitutions were implemented so that 47 patients initially prescribed intermittent infusion of piperacillin-tazobactam were switched to continuous infusion of this drug combination. Dosages varied in accordance with the type of infection and each patient's renal function. Fifty one other patients with similar demographics and types of infection received intermittent infusion with piperacillin-tazobactam. MEASUREMENTS AND MAIN RESULTS: Clinical success rates were 94% for the continuous-infusion group and 82% for the intermittent-infusion group (p=0.081). Microbiologic success rates were 89% for the continuous-infusion group and 73% for the intermittent-infusion group (p=0.092). Days to normalization of fever were significantly lower (p=0.012) in the continuous-infusion group (1.2 +/- 0.8 days) than in the intermittent-infusion group (2.4 +/- 1.5 days). Level 1 and level 2 costs/patient were both reduced by continuous infusion, although the difference was statistically significant only for level 2 costs ($399.38 +/- 407.22 for continuous infusion vs $523.49 +/- 526.85 for intermittent infusion, p=0.028). CONCLUSION: Continuous infusion of piperacillin-tazobactam provided clinical and microbiologic outcomes equivalent to those for intermittent infusion. Compared with intermittent infusion, continuous infusion significantly shortened the time to temperature normalization, while also offering a significant reduction in level 2 expenditures. PMID- 11939683 TI - Treatment of sickle cell pain. AB - Sickle cell disease affects 70,000 Americans who experience an average of 0.8 painful episodes each year. The pathophysiology of sickle cell pain is not completely understood. The disease is characterized by both acute and chronic pain syndromes. Patients with sickle cell pain often encounter barriers to receiving appropriate care, including lack of continuity of care and perceived opiate addiction. Studies describing pharmacotherapy for sickle cell pain have been primarily retrospective and uncontrolled. In analyzing the available literature regarding pathophysiology, assessment, and treatment of sickle cell pain, we found a need for increased practitioner education and intervention to improve the level of care provided to patients with this disease. PMID- 11939684 TI - Removal of lepirudin, a recombinant hirudin, by hemodialysis, hemofiltration, or plasmapheresis. AB - Lepirudin (recombinant hirudin), a direct thrombin inhibitor, is an effective alternative method of anticoagulation in patients with heparin-induced thrombocytopenia. However, because it is eliminated by the kidneys, the half-life of lepirudin may be substantially prolonged in patients with renal failure. Patients undergoing hemodialysis must be closely monitored, and therapy must be individualized based on each patient's ability to clear the drug. Current literature on the removal of lepirudin by dialysis or plasmapheresis is limited, but available data suggest that lepirudin can be removed with these methods. The ability of filtration systems to remove lepirudin from the blood is highly dependent on the membrane material used in the system. Understanding the effects of hemodialysis, hemofiltration, and plasmapheresis on lepirudin levels is important, especially since no antidote is available to treat elevated serum lepirudin concentrations. PMID- 11939685 TI - Patient perceptions of a proton pump inhibitor therapeutic interchange program across the continuum of care. AB - OBJECTIVE: To determine if participation in a hospital-based proton pump inhibitor (PPI) therapeutic interchange program resulted in differences in patient perceptions related to clinical and humanistic outcomes. METHODS: A quasiexperimental repeated-measures study compared patients' perceptions in two groups (111 patients) at hospital admission, discharge, 3-4 days after discharge, and 2-3 weeks after discharge to detect differences across the continuum of care. Patient awareness of the hospital-based interchange also was examined. Clinical and quality-of-life outcomes were measured by using a condition-specific instrument modified for use with patients. Satisfaction and expectations were measured by using extent-of-agreement measures. Group 1 (60 patients) was prescribed omeprazole before admission, switched to lansoprazole during hospitalization, and discharged taking omeprazole. Group 2 (51 patients) was prescribed lansoprazole before admission and continued taking lansoprazole throughout hospitalization and after discharge. Patients who were unable to communicate or who had a substantial change in severity of illness (not attributable to gastrointestinal disorders) during the study were excluded. RESULTS: No significant differences were found between groups 1 and 2 for clinical outcomes, quality of life, extent to which expectations were met, or satisfaction at the four time periods. Nineteen (36%) of 53 surveyed patients in group 1 were not aware that their therapy had been interchanged while in the hospital. CONCLUSION: Patient-perceived outcomes and expectations of therapy across the continuum of care were not affected by a hospital-based PPI therapeutic interchange program. PMID- 11939686 TI - Rabbit syndrome secondary to risperidone. AB - After taking risperidone for 4 months, a 38-year-old man with a major depressive disorder experienced rabbit syndrome. This uncommon extrapyramidal side effect is characterized by rhythmic movements of the mouth and may be caused by typical antipsychotics and risperidone secondary to blockade of dopamine D2 receptors. The syndrome, whose exact mechanism is unknown, responded to anticholinergic treatment in this patient. A literature review indicated that most recent cases have occurred as reactions to the atypical antipsychotic risperidone. Changing treatment to an atypical antipsychotic other than risperidone, such as olanzapine 10 mg/day, may lead to suppression or elimination of the syndrome. PMID- 11939687 TI - Selective serotonin reuptake inhibitor-induced rash: case report and review of the literature. AB - Selective serotonin reuptake inhibitors (SSRIs) are one of the most frequently prescribed classes of drugs. Rashes induced by SSRIs seldom have been reported in the literature. Computerized MEDLINE and Current Contents searches yielded a report of two cases of rash induced specifically by paroxetine. We describe a patient who developed a rash that appeared on day 3 of treatment with oral paroxetine 20 mg/day. Her rash was morbilliform, pruritic, and generalized over the trunk and limbs, with some facial involvement. There was no palm or sole involvement. The patient reported that she had experienced a similar reaction to fluoxetine, which may suggest a drug class effect. The rash resolved within 2 days of drug discontinuation and treatment with oral diphenhydramine and topical hydrocortisone cream. PMID- 11939688 TI - Rhinocerebral mucormycosis in the era of lipid-based amphotericin B: case report and literature review. AB - Rhinocerebral mucormycosis (RCM) is an invasive fungal infection that necessitates, in most cases, aggressive surgical debridement and high cumulative, often nephrotoxic doses of amphotericin B. A 50-year-old woman with RCM was treated successfully with amphotericin B lipid complex as primary therapy. The patient previously had displayed progressive intracranial involvement and rising serum creatinine levels while receiving the conventional (nonlipid) form of amphotericin B. A literature review identified only a few cases where systemic antifungal therapy was administered, with minimal or no surgery. Our case further supports that amphotericin B lipid complex can be used as primary therapy in selected patients with RCM, without the need for surgical exenteration. PMID- 11939689 TI - Nontraditional dosing of ampicillin-sulbactam for multidrug-resistant Acinetobacter baumannii meningitis. AB - A 52-year-old man was admitted to a local hospital with headache, nausea, vomiting, dizziness, photophobia, and confusion after a sudden fall. Progressive changes in neurologic function were noted despite neurosurgical intervention and broad-spectrum antimicrobial coverage. Cerebral spinal fluid (CSF) culture identified Acinetobacter baumannii that was resistant to traditionally recommended therapies of amikacin and imipenem-cilastatin. The organism demonstrated minimum inhibitory concentrations of greater than 32 microg/ml and 8 microg/ml, respectively, for these two agents. Ampicillin 2 g-sulbactam 1 g every 3 hours was administered based on history of therapeutic failure of traditional dosing in our thermal injury population. Repeat CSF cultures after 12 days of ampicillin-sulbactam therapy were negative. After 35 days, the patient's A. baumannii infection was completely resolved. The patient experienced no adverse drug events or toxicity with this high-dosage regimen. PMID- 11939690 TI - Cerivastatin-induced rhabdomyolysis: 11 case reports. AB - 3-Hydroxy-3-methylglutaryl coenzyme A (HMG-CoA) reductase inhibitors are a frequently prescribed class of drugs that are generally well tolerated by most patients. A rare, yet serious side effect associated with these drugs is rhabdomyolysis. Although all HMG-CoA reductase inhibitors can cause this adverse effect, prevalence may differ among specific agents. Over a 16-month period, in three hospitals, 11 patients experienced cerivastatin-induced rhabdomyolysis, but no cases of rhabdomyolysis associated with any other HMG-CoA reductase inhibitors were reported. PMID- 11939691 TI - Citalopram-induced priapism. AB - Citalopram is a relatively new selective serotonin reuptake inhibitor (SSRI) that is becoming widely administered for the treatment of depression. Selective serotonin reuptake inhibitors generally are associated with mild adverse sexual side effects; however, more serious reactions may occur. A 58-year-old man experienced priapism several hours after inadvertently taking three tablets of citalopram 20 mg, which he had mistaken for aspirin, in addition to his usual dosage of 20 mg twice/day. Three days later, he was hospitalized and treated with intracavernous phenylephrine. He ultimately required surgical intervention. Although the citalopram overdose appears to be largely responsible for the patient developing priapism, he also was taking tamsulosin 0.4 mg/day at bedtime for benign prostatic hyperplasia. As alpha1-blockers have been associated with priapism on rare occasions, tamsulosin may have been a contributing factor. The patient also had a history of priapism associated with trazodone. Health care professionals should vigilantly monitor patients who take citalopram in high dosages or in combination with other drugs associated with priapism. Patients who have a history of priapism with other drugs may be more susceptible to citalopram associated priapism. PMID- 11939692 TI - Superiority of enoxaparin over unfractionated heparin for the treatment of acute coronary syndromes. PMID- 11939694 TI - Topography of somatosensory processing: cerebral lateralization and focused attention. AB - Healthy dextrals underwent fMRI during a task of graphesthesia requiring detection of any number written consecutively from an otherwise random number sequence. Test conditions included (1) focus on unilateral right hand stimuli, (2) focus on unilateral left hand stimuli, (3) focus on right hand only during bilateral hand stimulation, (4) focus on left hand only during bilateral hand stimulation, and (5) rest. Attention to unilateral hand stimulation produced bihemispheric activation with minimal or no activation of ipsilateral primary sensorimotor region. Attention to unilateral left hand stimuli resulted in more activation than attention to unilateral right hand stimuli. Stimulation of the nonattended hand activated the contralateral somatosensory area, but to a lesser spatial extent than attended stimuli. Comparing focused attention to the left versus right side during identical sensory inputs (i.e., bilateral hand stimulation), focused attention to the right hand increased activation in the left somatosensory region, but focused attention to the left hand increased activation in both cerebral hemispheres. Thus, focused attention to unilateral somatosensory stimuli produces bilateral cerebral activation, but the increase in blood flow is greater in the contralateral hemisphere. Unattended stimuli activate the contralateral primary somatosensory area. Left/right asymmetries were demonstrated consistent with cerebral lateralization. PMID- 11939693 TI - Reading level attenuates differences in neuropsychological test performance between African American and White elders. AB - The current study sought to determine if discrepancies in quality of education could explain differences in cognitive test scores between African American and White elders matched on years of education. A comprehensive neuropsychological battery was administered to a sample of African American and non-Hispanic White participants in an epidemiological study of normal aging and dementia in the Northern Manhattan community. All participants were diagnosed as nondemented by a neurologist, and had no history of Parkinson's disease, stroke, mental illness, or head injury. The Reading Recognition subtest from the Wide Range Achievement Test-Version 3 was used as an estimate of quality of education. A MANOVA revealed that African American elders obtained significantly lower scores than Whites on measures of word list learning and memory, figure memory, abstract reasoning, fluency, and visuospatial skill even though the groups were matched on years of education. However, after adjusting the scores for WRAT-3 reading score, the overall effect of race was greatly reduced and racial differences on all tests (except category fluency and a drawing measure) became nonsignificant. These findings suggest that years of education is an inadequate measure of the educational experience among multicultural elders, and that adjusting for quality of education may improve the specificity of certain neuropsychological measures. PMID- 11939695 TI - Increased variability accompanies frontal lobe damage in dementia. AB - Performance variability on neuropsychological measures is not a unitary phenomenon, and different measures (consistency, dispersion, diversity) evaluate separate elements of variability. It has been suggested that increased variability may be a specific attribute of frontal lobe pathology. This hypothesis was tested in 2 matched groups of demented subjects, 8 with dementia of the Alzheimer type (DAT), 5 with frontal lobe dementia (FLD), compared with 10 elderly normal controls (ENC). A Stroop test and Reaction Time measures were administered weekly for 5 weeks to all subjects. Both measures contained three subtests varying in degree of complexity. The results from the Stroop task indicated that the FLD group showed significantly greater variability on measures of consistency (fluctuations over time) and diversity (between participant variability) regardless of the complexity of the subtest. For the Reaction Time subtests, measures of consistency and diversity showed significantly greater variability in FLD, but were affected in a different pattern. Greater variability in terms of consistency of performance was manifested only in the more attentionally demanding of the Reaction Time subtests (Choice Reaction Time, CRT). On the measure of diversity, variable performance was found to be greater on the Simple Reaction Time (SRT) subtest than on the more effortful CRT. Dispersion (within participant variability) was only assessed on the reaction time subtests. The results indicate no significant evidence for an increase in dispersion for the FLD patients. The hypothesis that variability will be increased in frontal lobe dementia is thus confirmed, and the independence of the three forms of variability measurement is demonstrated in dementia subjects. PMID- 11939696 TI - Apathy and executive function in Alzheimer's disease. AB - Apathy is a common behavioral disturbance in patients with Alzheimer's disease (AD). Recent studies have linked the presence of apathy to alterations in frontal lobe functions, but few studies have explored the relationship using standard neuropsychological measures in patients with AD. We administered a comprehensive battery of neuropsychological tests and a behavior rating scale to 80 patients with AD. We explored the relationship of apathy to executive dysfunction. AD patients with apathy performed significantly worse on tests of executive function (WAIS-R Digit Symbol, Trail-Making, Stroop Color Interference Test) than AD patients without apathy. The presence of dysphoria did not modify these results and no significant relationships were found between tests of executive functions and dysphoria. Performance on executive measures as a group were effective in correctly classifying patients as apathetic or nonapathetic with 75% accuracy. Neuropsychological measures not dependent on executive functions were unrelated to apathy. Apathy is associated with executive dysfunction and not with other neuropsychological deficits. Apathy is distinct from dysphoria. PMID- 11939697 TI - Search pattern in a verbally reported visual scanning test in patients showing spatial neglect. AB - The present study of right hemisphere stroke patients showed that presence of visuospatial neglect in conventional neglect tests at the postacute stage was strongly associated with an aberrant search pattern in a verbally reported visuo perceptual scanning test. Compared with normal controls, patients with visuospatial neglect showed a greater proportion of repeated readings of the same target, shorter search sequences, more shifts between horizontal, vertical, and diagonal search, and lower proportion of horizontal search. The relation between spatial neglect and a deficient search pattern was strongly influenced by the asymmetric allocation of attention in the scanning test, with the exception for the proportion of repeated reading which was not influenced by this asymmetry. At follow-up, a significant recovery was noted in the neglect group for the proportion of repeated readings and for the asymmetry in the allocation of attention. However, a high number of omitted targets in the search test was still a common finding in the neglect group and it was suggested that a non-lateralized attentional deficit may have played an important role behind the ineffective search at this point of time. PMID- 11939698 TI - Memory in multiple sclerosis: contextual encoding deficits. AB - Long-term memory (LTM) is one of the diverse cognitive functions adversely affected by multiple sclerosis (MS). The LTM deficits have often been attributed to failure of retrieval, whereas encoding processes are presumed intact. However, support for this view comes primarily from studies in which encoding and retrieval operations were not investigated systematically. In the current study, we used an encoding specificity paradigm to examine the robustness of encoding in MS and to specifically evaluate the impact of the disease on contextual memory. We hypothesized that persons with MS would exhibit a selective impairment in retrieving items from LTM when required to generate new cue-target associations at encoding, but not when cues held a strong preexisting relationship to the targets. The findings supported the hypotheses. We conclude that the mnemonic deficits associated with MS affect both encoding and retrieval. Specifically, problems with binding of contextual information at encoding impair effective retrieval of memories. Nonetheless, access to these memories can be gained through preexisting associations organized in the semantic network. PMID- 11939699 TI - A meta-analysis of the neuropsychological sequelae of HIV infection. AB - This meta-analysis summarizes the broad spectrum of neuropsychological research on HIV disease across a sample of 41 primary studies and an aggregate of 8,616 participants for 10 major neuropsychological ability areas. Analyses of the course of cognitive decline within and across Centers for Disease Control classifications reveals statistically significant cognitive deficits from asymptomatic HIV to AIDS. Effect sizes (Cohen, 1988) were calculated to reflect between-group (asymptomatic, symptomatic, AIDS) differences in each neuropsychological domain. Relatively small effect sizes were obtained for the asymptomatic (0.05-0.21) patients, and generally small to moderate effect sizes were obtained for symptomatic (0.18-0.65) HIV+ patients, with motor functioning exhibiting the greatest effects in this later disease stage. The most notable deficits in cognitive functioning were found in the AIDS group with moderate (attention and concentration) to large (motor functioning) effect sizes with values ranging from 0.42-0.82. Comparison of cognitive functioning as a function of disease progression revealed that motor functioning, executive skills, and information processing speed were among the cognitive domains showing the greatest decline from early to later stages of HIV. These findings indicate that cognitive deficits in the early stages of HIV are small and increase in the later phases of the illness, and that specific patterns of cognitive deficits can be detected with disease progression. These results and their clinical utility are further discussed. PMID- 11939700 TI - New semantic and serial clustering indices for the California Verbal Learning Test-Second Edition: background, rationale, and formulae. AB - The original California Verbal Learning Test (CVLT) employed a semantic clustering index that used the words recalled during a given trial as the baseline for calculating expected values of chance clustering (recall-based expectancy). Although commonly used in cognitive psychology, clustering indices that use recall-based calculations of expectancy are implied by the assumption that organizational processes do not occur until after words are retrieved from memory. This assumption contradicts the generally held assumptions among neuropsychologists that (1) organization is an antecedent to recall, and (2) increases in the use of organizational strategies will result in better recall performance. After reviewing a brief history of clustering metrics, we used Monte Carlo simulations, informative examples, and patient data to examine clustering indices that use the word list as a baseline for calculating expectancy and propose these list-based expectancy measures as a refinement of the clustering indices used on the original CVLT. These indices are used on the recently published CVLT-II. PMID- 11939701 TI - Social support, neuropsychological performance, and depression in HIV infection. AB - The present study was designed to examine the impact of neuropsychological performance on the relationships between stress, social support, and depression in 217 HIV-infected men. Using path analysis, the contributions of four domains of cognitive functioning (memory, attention, executive function, and psychomotor speed), IQ, and relevant psychosocial variables to depression were evaluated. In the model which best fit the data, cognitive domains did not contribute directly to depression, but contributed significantly to psychosocial variables which affected levels of depression. Attention and executive function contributed to reduced illness-related disability; while higher IQ was associated with fewer stressful life events. Number of stressful life events and level of illness related disability were associated with depressive symptoms. Higher IQ led to greater numbers of social contacts, which was associated with fewer reported symptoms of depression. These findings suggest that better neuropsychological performance may lead to reduced stress and perceived disability, and more available social contacts. By these multiple paths, different domains of cognitive ability contribute indirectly to ameliorating depression in HIV infected men. PMID- 11939702 TI - What is cognitive reserve? Theory and research application of the reserve concept. AB - The idea of reserve against brain damage stems from the repeated observation that there does not appear to be a direct relationship between the degree of brain pathology or brain damage and the clinical manifestation of that damage. This paper attempts to develop a coherent theoretical account of reserve. One convenient subdivision of reserve models revolves around whether they envision reserve as a passive process, such as in brain reserve or threshold, or see the brain as actively attempting to cope with or compensate for pathology, as in cognitive reserve. Cognitive reserve may be based on more efficient utilization of brain networks or of enhanced ability to recruit alternate brain networks as needed. A distinction is suggested between reserve, the ability to optimize or maximize normal performance, and compensation, an attempt to maximize performance in the face of brain damage by using brain structures or networks not engaged when the brain is not damaged. Epidemiologic and imaging data that help to develop and support the concept of reserve are presented. PMID- 11939704 TI - Progress toward global eradication of poliomyelitis, 2001. AB - From the initiation of the global poliomyelitis eradication initiative in 1988 through 2001, the number of countries where polio is endemic decreased from 125 to 10, and the number of reported polio cases decreased by >99% from an estimated 350,000 to <1,000. Wild type 2 poliovirus has not been detected worldwide since October 1999. The American and Western Pacific Regions of the World Health Organization (WHO) have been certified free of indigenous wild poliovirus. Current challenges to global polio eradication efforts include ongoing intense transmission in northern India, continued importations of wild poliovirus into polio-free areas, and the detection of circulating vaccine-derived poliovirus (cVDPV). This report summarizes global progress in polio eradication during 2001 and the current status of the initiative. PMID- 11939703 TI - Visual perception without awareness in a patient with posterior cortical atrophy: impaired explicit but not implicit processing of global information. AB - A patient with progressive posterior cortical atrophy (PCA) was examined on several tests of visual cognition. The patient displayed multiple visual cognitive deficits, which included problems identifying degraded stimuli, attending to two or more stimuli simultaneously, recognizing faces, tracing simple visual stimuli, matching simple shapes, and copying objects. The patient was also impaired in identifying visual targets contained at the global level within global-local stimuli (i.e., smaller letters that compose a larger letter). Although the patient denied any conscious awareness of the global form, he nevertheless displayed a normal pattern of global interference when asked to identify local level targets. Thus, the patient processed the global information despite not being consciously aware of such information. These results suggest that global-local processing can take place in the absence of awareness. Possible neurocognitive mechanisms explaining this dissociation are discussed. PMID- 11939705 TI - Reporting of laboratory-confirmed chlamydial infection and gonorrhea by providers affiliated with three large managed care organizations--United States, 1995-1 999. AB - Surveillance for sexually transmitted diseases (STDs) depends on health departments receiving reports of positive STD test results from laboratories or of STD cases by clinicians. The completeness and timeliness of reporting can affect prompt sex partner notification and outbreak detection. In 1998, approximately 70% of chlamydia cases and 55% of gonorrhea cases were reported by private clinicians, including many affiliated with managed care organizations (MCOs). However, little is known about the completeness and timeliness of MCOs' STD case reporting practices. Three MCOs, three state health departments, and CDC evaluated reporting practices for chlamydial infection and gonorrhea by three large staff or group model MCOs that used different reporting procedures. The findings indicate that state health departments were notified of 78%-98% of chlamydia cases and of 64%-80% of gonorrhea cases identified in these MCOs; the median interval between specimen collection and state health department receipt of a case report was < or = 19 days. To improve surveillance quality, other MCOs, including network model MCOs, which provide most STD care in the United States, should evaluate surveillance quality and identify interventions for improvement. PMID- 11939706 TI - Oral health survey of third grade students--New Hampshire, 2001. AB - Dental caries is one of the most common chronic disease of childhood. Despite improvements in oral health in the United States, by late adolescence approximately 80% of children have a history of caries. To establish a baseline for monitoring oral disease trends in New Hampshire, the New Hampshire Department of Health and Human Services in cooperation with the New Hampshire Department of Education conducted a statewide assessment of the oral health status of third grade students attending public schools during the 2000-2001 academic year. This report summarizes the findings of the survey, which indicate that approximately one half of third grade students in New Hampshire have experienced dental decay. Increasing access to fluoridated water and dental sealants is needed to prevent childhood caries. PMID- 11939707 TI - Cancer risk associated with household exposure to chloroform. AB - Chloroform (CHCl3) the trihalomethane most prevalent in drinking water, is a proven animal carcinogen and a suspected human carcinogen. Consequently, standards have been issued by health authorities to limit its concentration in drinking water. These limits are based solely on ingestion, without taking into account inhalation and skin contact. Exposure to CHCl3 was assessed for 18 men (age: mean 38 years; range 23-51) following a 10-min shower in their respective residences located in the Quebec City region (Canada). CHCl3 concentration was measured in alveolar air samples collected before, immediately after, and 15 min and 30 min following the shower. Indoor air and water concentrations were determined concomitantly. Mean CHCl3 concentrations in the air of the shower stall and in water were respectively 147 microg/m3 (SD = 56.2 microg/m3) and 20.1 microg/L (SD = 9.0 microg/L). Water concentrations were comparable to those documented in a large proportion of distribution networks in Canada. The mean increase in alveolar air CHCl3 concentration (deltaCHCIALV) at the end of the shower was 33 microg/m3 (SD = 14.7 microg/m3). A multiple-regression analysis revealed that deltaCHCl3ALV values were only associated with chloroform concentration in air of the shower stall. DeltaCHCl3ALV were described using a physiologically based pharmacokinetic (PBPK) model. This model was then used to estimate concentrations of CHCl3 metabolites bound to liver and kidney macromolecules following a shower, and also according to exposure scenarios that integrate drinking-water ingestion and air inhalation. The concentration predicted in the liver following a worst-case exposure scenario was 0.41 microg CHCl3 equivalents/kg of tissue, some 6,000 times lower than the lowest concentration that did not increase the incidence of hepatic tumors in laboratory animals. Data indicate that for this range of exposure the safety margin appears therefore considerable with respect to the potential carcinogenic effect of household exposure to CHCl3. PMID- 11939708 TI - Preliminary assessment of atmospheric methylcyclopentadienyl manganese tricarbonyl and particulate manganese in selected urban sites. AB - Methylcyclopentadienyl manganese tricarbonyl (MMT: C9H7MnO3) is an organometallic additive that has been used since 1976 as an octane enhancer in Canadian unleaded gasoline. Very few studies have determined its atmospheric concentrations and only one study offers recent data on its ambient level. This preliminary study aims to assess atmospheric concentrations of MMT and respirable and total Mn (Mn(R) and Mn(T) in selected sites, at two underground car parks and one gasoline station, related to high levels of automobile traffic. It is also an investigation of the applicability of the current analytical method. In total, 34 air samples were collected using a Gil-Air portable pump during 4 consecutive days and then were analyzed by neutron activation analysis. The concentrations vary between 40 and 104 ng/m3 for Mn(R), 146 and 204 ng/m3 for Mn(T) and 6 and 128 ng/m3 for MMT (including ultrafine particulates, Mn(UF). Of the 12 Mn(R) results, 7 showed concentrations greater than the U.S. EPA reference concentration (RfC = 50 ng/m3). The ratios of Mn(R) to Mn(T) varied from 0.20 to 0.65 with a mean of 0.38. The results for MMT and Mn(UF) raise serious doubts about the specificity of the sampling and chemical analysis methodology proposed by the Occupational Safety and Health Administration (OSHA) for MMT in air. PMID- 11939709 TI - Alterations of serum hormone levels in male workers occupationally exposed to cadmium. AB - In order to investigate the influence of occupational exposure to cadmium (Cd) on the levels of serum hormone, a cross-sectional study was undertaken in male workers. Creatinine-adjusted urinary cadmium (UCd) was used as the indicator of cadmium body burden in workers. The levels of serum testosterone (T), follicle stimulating hormone (FSH), and luteinizing hormone (LH) were determined by radioimmunoassay and enzyme immunoassay, respectively. The results showed that with cadmium exposure, the levels of serum testosterone and LH were significantly increased after controlling for confounding factors such as age, smoking habits, and alcohol consumption. However, no significant cadmium-associated changes in the levels of serum FSH were found. At the same time, a significant negative correlation of serum testosterone levels with age was detected. In contrast, the levels of serum FSH and LH were significantly elevated with increasing age. When taking the marital status into consideration, only significantly elevated FSH levels were noted in married workers after controlling for confounding factors. These results indicate that occupational cadmium exposure and other factors such as age and marital status may independently contribute to alterations in the levels of serum sex hormones in males. PMID- 11939710 TI - Effect of methacrylonitrile on cytochrome P-450 2E1 (CYP2E1) expression in male F344 rats. AB - Tissue-specific induction of cytochrome P-450s (CYP) followed by increased in situ bioactivation may contribute to chemical-induced site-specific toxicity. In rats, methacrylonitrile (MAN) is metabolized by cytochrome P-450 2E1 (CYP2E1) to acetone, which is eliminated along with parent MAN in breath. Gavage administration of MAN to rats causes olfactory epithelial damage and liver enlargement. It was hypothesized that treatment of rats with MAN may result in differential expression of CYP2E1 in tissues leading to tissue-specific toxicity via increased in situ formation of cytotoxic MAN metabolites. In this study, male F344 rats received 60 mg MAN/kg and were sacrificed 6, 12, or 24 h after a single dose, or 24 h after 7 consecutive daily doses. Liver, lung, and nasal tissues were collected. Reverse-transcription polymerase chain reaction (RT-PCR), Western blotting, and immunohistochemical staining were used to assess CYP2E1 expression and localization, and chlorzoxazone hydroxylation was used as a measure of CYP2E1 catalytic activity. Present results showed that CYP2E1 mRNA was increased in lung and nasal tissues with minimal effects in liver of MAN-treated rats. Induction of CYP2E1 protein expression was detected in lung. CYP2E1 activity was higher in liver and lung microsomes from MAN-treated rats when compared to control animals. To compare the effects of MAN and acetone, male F344 rats received a single acetone dose (5 ml/kg) by gavage. After 12 h, acetone treatment resulted in a significant increase in the levels of CYP2E1 mRNA and protein in lung and nasal tissues, with no obvious change noted in the liver. Overall, these data suggest that administration of MAN to rats causes increased expression of CYP2E1 in lung, liver, and nasal tissues. These results also show that acetone induces the expression of CYP2E1 at both the mRNA and protein levels in rat nasal and lung tissues. In conclusion, MAN increased the expression of CYP2E1, and this effect varied as a function of time, length of exposure, and tissue examined. While the damage in the olfactory mucosa due to MAN treatment may not be explained by the observed induction of CYP2E1, it is possible that other CYPs may play a role in the in situ bioactivation of MAN. PMID- 11939711 TI - Effect of three n-acetylamino acids on N-(3,5-dichlorophenyl)succinimide (NDPS) and ndps metabolite nephrotoxicity in Fischer 344 rats. AB - The agricultural fungicide N-(3,5-dichlorophenyl)succinimide (NDPS) induces nephrotoxicity in mammals characterized as polyuric renal failure and proximal tubular necrosis. Recent studies have suggested that NDPS-induced nephrotoxicity may be mediated by metabolites arising from the nephrotoxic NDPS metabolites N (3,5-dichlorophenyl)-2-hydroxysuccinimide (NDHS) and/or N-(3,5-dichlorophenyl)-2 succinamic acid (2-NDHSA). The purpose of this study was to examine the effects of N-acetylcysteine (NAC), a nucleophilic agent, and two nonnucleophilic N acetylamino acids, N-acetylserine (NAS) and N-acetylalanine (NAA), on NDPS and NDPS metabolite-induced nephrotoxicity. Male Fischer 344 rats (4-8/group) were administered intraperitoneally (ip) an N-acetylamino acid (1 mmol/kg) 2 h before an ip injection of NDPS (0.4 mmol/kg), NDHS (0.1 mmol/kg), 2-NDHSA (0.1 mmol/kg), or vehicle. Renal function was then monitored at 24 and 48 h. NAC pretreatment markedly attenuated NDPS-, NDHS-, and 2-NDHSA-mediated nephrotoxicity. The nonnucleophilic N-acetylamino acids (NAS, NAA) only partly reduced NDPS and NDHS nephrotoxicity, and they had little effect on 2-NDHSA nephrotoxicity. These results suggest that reactive NDPS metabolites may be formed from NDHS and 2 NDHSA and that nucleophilic substrates (e.g., NAC) may offer protection from NDPS induced nephrotoxicity. However, mechanisms other than chemical neutralization of reactive NDPS metabolites may also be contributing to the attenuation of NDPS nephrotoxicity, since nonnucleophilic N-acetylamino acids (e.g., NAA) also provided some protection against NDPS and NDHS nephrotoxicity. PMID- 11939712 TI - Concentration dependency in lead-inhibited steroidogenesis in MA-10 mouse Leydig tumor cells. AB - Lead acetate (Pb) decreases the expression of steroidogenic acute regulatory (StAR) protein and the enzymatic activities of cytochrome P-450 side-chain cleavage (P450scc) and 3beta-hydroxysteroid dehydrogenase (3beta-HSD) in a concentration-dependent manner in Leydig cells at 2 h, the duration of submaximal inhibition. This study was undertaken at 3 h of Pb incubation to compare the effects at maximal metal inhibition of steroidogenesis. Quantitatively a 3-h Pb incubation with MA-10 cells resulted in higher decreases in human chorionic gonadotropin (hCG)-stimulated progesterone production, expression of StAR protein, and the activity of 3beta-HSD compared to 2 h. In contrast, lead inhibited dibutyryl cAMP (dbcAMP)-stimulated progesterone production but lacked this effect at 2 h. Surprisingly, Pb at 3 h of incubation did not affect P450scc enzyme activity, yet this enzymatic activity was inhibited at 2 h. Data indicate that incubation time is a factor in Pb-induced alterations in MA- 10 cell steroidogenesis. PMID- 11939713 TI - Effect of repeated treatment with mirtazapine on the central alpha1-adrenergic receptors. AB - Mirtazapine (MIR) is an antidepressant which enhances noradrenergic and serotonergic 5-HT1A neurotransmission via antagomism of central alpha2-adrenergic autoreceptors and heteroreceptors. The drugs does not inhibit noradrenaline and serotonin reuptake but blocks the 5-HT, and 5-HT3 receptors and has high affinity only for central and peripheral histamine H1 receptors. The present study was aimed at determining whether repeated MIR treatment induced adaptive changes in the alpha1-adrenergic receptors, similar to those reported by us early for tricyclic antidepressants, The experiments were carried out on male mice and rats. MIR was administered at a dose of 10 mg/kg once or repeatedly (twice daily for 14 days). The obtained results showed that MIR administrated repeatedly potentiated the methoxamine- induced exploratory hyperactivity in rats and clonidine-induced aggressiveness in mice, those effects being mediated by alpha1 adrenergic receptors. MIR given repeatedly (but not acutely) increased the binding (Bmax ) of [3H]prazosin to alpha1-adrenergic receptors in cerebral cortex, however, the ability of the alpha1-adrenoceptor agonist phenylephrine to compete for the these sites was not significantly changed. The above results indicate that repeated MIR administration increases the responsiveness of alpha1 adrenergic system (behavioural and biochemical changes), as tricyclics do. However, the question whether the increased functional responsiveness found in the present study is important for the clinical antidepressant efficacy, remains open. PMID- 11939714 TI - Effect of tianeptine and fluoxetine on the levels of Met-enkephalin and mRNA encoding proenkephalin in the rat. AB - The purpose of the present study was to evaluate the effects of acute and repeated treatment with two antidepressant drugs (ADs) of opposite pharmacological profile, i.e. tianpetine (TIA, serotonin reuptake enhancer) and fluoxetine (FLU, serotonin reuptake inhibitor) on the levels of Met-Enkephalin, (Met-Enk, a member ofopioid peptide family, which has been suggested to play a role in the mechanism of action ADs) as well as on mRNA coding for proenkephalin (mRNA PENK) in various regions of the rat brain, pituitary, adrenal glands and plasma. Male Wistar rats were treated acutely or repeatedly (10 mg/kg p.o., twice daily for 14 days) with TIA or FLU. Tissue for biochemical experiments was taken 2 h after last dose of appropriate drug. The levels of Met-Enk were estimated by radioimmunoassay, mRNA PENK was measured using in situ hybridization. From the results obtained in the present study it may be concluded that repeated administration of TIA or FLU induced similar changes in the levels of Met-Enk in the rat hippocampus, striatum, hypothalamus and neurointermediate lobe of pituitary. Such an effect is interesting, especially if one takes into account the differences in pharmacological profile between these two antidepressant drugs. It may be suggested that serotonin level might not be crucial for inducing the alterations in the content of Met-Enk. Since we did not observe any changes in the levels of PENK mRNA in the studied rat brain regions after repeated administration of TIA or FLU, it seems that the observed changes in the levels of Met-Enk do not result from effects of these antidepressants on biosynthesis of PENK, but rather from alterations in the peptide release. Another interesting finding of the present study was that in the anterior lobe of pituitary, adrenal glands and plasma, repeated administration of TIA induced alterations in the contents of Met-Enk, while repeated administration of FLU remained without any effect. It is tempting to speculate that such a differentiation between the effects of these two antidepressants might be linked to the well known feature of TIA (but not FLU) which has been shown to reduce both basal and stress-evoked activity of the hypothalamic-pituitary-adrenal (HPA) axis. PMID- 11939715 TI - Eosinophil-epithelial cell interaction augments cysteinyl leukotrienes synthesis. AB - Eosinophils accumulation in the airways and sustained eosinophil-derived cysteinyl leukotrienes production represent key elements of the inflammatory response seen in asthma. However, it is not known whether activated epithelial cells influence cysteinyl leukotrienes production by eosinophils from healthy valunteers. The aim of the present study was therefore to analyse the effects of interactions between non-atopic eosinophils and epithelial cells on cysteinyl leukotrienes production in vitro. We measured cysteinyl leukotrienes released by phorbol 12-myristate 13-acetate (PMA) -activated human eosinophils or epithelial cells (human bronchial epithelial cell line -BEAS-2B) cultured alone or together. While activated BEAS-2B cells barely formed leukotrienes (1.39 pg/ml +/- 0.2) (n=32), activated eosinophils produced considerable amount of them (62.25 pg/ml +/- 10.29) (n=32). Interestingly, when activated eosinophils and epithelial cells were co-incubated, production of cysteinyl leukotrienes increased substantially (571.1 pg/ml +/- 80.9) (n=32). Thus, eosinophil-epithelial cell interactions, when occur, are associated with increased biosythesis of cysteinyl leukotrienes. PMID- 11939716 TI - Screening and modulation of extracellular signals by mucous barrier. Serum glycosylphosphatidylinositol phospholipase D (GPI-PLD) releases protective mucous barrier from oral mucosa. AB - BACKGROUND: Performance of mucosal epithelial barrier is modified by numerous agents that exert effects on mucin- Mucin Binding Protein (MBP) complex. The aim ofthe studies described was to determine the nature of the damage or modification of oral mucous barrier by the short-term exposure to ethanol. METHODS: Culture of rat buccal mucosa in the presence of ethanol and [3H]-labeled proline and palmitate revealed substantial decrease in MBP synthesis and the release of MBPto the medium. The radioscanning of the samples prepared from the culture medium and the apical epithelial membranes subjected to SDS-PAGE and western blotting disclosed that the released, water soluble 97kDa MBP glycopeptide was labeled with proline and palmitate. When the experiments were conducted in the presence of 5mM EDTA, the GPI-PLD inhibitor, the majority of radiolabeled MBP remained in the membrane-bound form and was extractable with Triton X- 114. The results on the purified GPI-linked MBP degradation by serum enzyme, by the saliva containing serum transudate, and the suppression of the process by inclusion of GPI-PLD specific inhibitor support our contention that membrane MBP is released to medium by GPI-PLD-like activity. RESULTS: The release of MBP from apical epithelial surfaces was induced by depletion of mucin and the presence of serum-derived GPI PLD in the tissue homogenate. Strong likelihood exists that under in situ conditions ethanol-induced transudation of serum to saliva provides the vehicle for the transfer of GPI-PLD activity to salivary contents. Defacement of the oral surfaces from mucous barrier signals prospect of lumenal agent influence on the unprotected epithelial exterior, and allows ingression of microbes and untoward acting substances into the organism. PMID- 11939717 TI - PEPCK mRNA localization in proximal tubule and gene regulation during metabolic acidosis. AB - To identify the nephron segments expressing PEPCK in control and acidotic conditions, PEPCK mRNA was localized in rat kidney using the technique of reverse transcription and polymerase chain reaction (RT-PCR) in individual microdissected S1 S2, and S3 segments of the rat proximal tubule. In controls, the number of tubules expressing PEPCK mRNA was greatest in the S3 segment, moderate in the S2 segment, and least in the S1 segment of the proximal tubule. After NH4Cl feeding, strong signals for PEPCK mRNA were detected in all three proximal tubule segments. In situ hybridization demonstrated expression of PEPCK mRNA only in the medullary rays in controls. After NH4Cl, PEPCK mRNA was expressed throughout the cortex, confirming the RT-PCR results. These data demonstrate the ability of the rat kidney cortex to modulate the expression of PEPCK mRNA during metabolic acidosis by recruitment of additional cells in the proximal nephrons. Studies with cultured LLC-PK1-F+ cells indicated that increased PEPCK gene transcription at acid pH required a cis-acting element (enhancer) in the more distal 5' flanking region of the promoter. PMID- 11939718 TI - Effects of reactive oxygen species action on gastric mucosa in various models of mucosal injury. AB - BACKGROUND: The exposure of gastric mucosa to damaging factors, such as ethanol, water restraint stress, or ischemia followed by reperfusion, produces pathological changes: inflammatory process, hemorrhagic erosions, even acute ulcers. The base of these changes is a disturbance of protective mechanisms and disrupture of gastric mucosal barrier. Previous studies pointed out the role of disturbances of gastric blood flow, mucus secretion and involvement of prostaglandins and nitric oxide formation in the pathomechanism of gastric mucosa lesions. The role of reactive oxygen species (ROS) in these processes has been little studied. AIM: The purpose of our present investigations is to explain the participation of ROS in acute gastric mucosal damage by various irritants. MATERIAL AND METHODS: Experiments were carrying out on 80 male Wistar rats. To assess gastric blood flow (GBF) laser Doppler flowmeter was used. The area of gastric lesions was established by planimetry. The levels of proinflammatory cytokines were measured by ELISA technique. The colorimetric assays were used to determine of malondialdehyde (MDA) and 4-hydroxynonenal (4-HNE) as well as superoxide dismutase (SOD) activity. RESULTS: We demonstrated that 3.5 h of water immersion and restraint stress (WRS), 30 min of gastric ischemia followed by 60 min of reperfusion or intragastric administration of 100% ethanol, all resulted in appearance of acute gastric mucosal lesions accompanied by a significant decrease of gastric blood flow. These lesions are also accompanied by the significant increase of proinflammatory cytokines including interleukin-1 beta (IL-1beta) and tumor necrosis factor alpha (TNFalpha) plasma level. Biological effects of ROS were estimated by measuring tissue level of MDA and 4-HNE, the products of lipid peroxydation by ROS, as well as the activity of SOD, the scavanger of ROS. It was established that 3.5 h of WRS, ischemia-reperfusion and 100% ethanol lead to significant increase of MDA and 4-HNE mucosal level, accompanied by a decrease of SOD activity (significant in WRS and ethanol application). CONCLUSIONS: The pathogenesis of experimental mucosal damage in rat stomach includes the generation of ROS that seem to play an important role, namely due to generation of lipid peroxides, accompanied by impairment of antioxidative enzyme activity of cells. PMID- 11939720 TI - Central histamine-induced reversal of critical haemorrhagic hypotension in rats- haemodynamic studies. AB - Volume-controlled irreversible haemorrhagic shock in rats produced by blood withdrawal until stabilisation of critical mean arterial pressure (MAP) 20-25 mmHg is associated with an extreme decrease in cardiac index (CI) and an increase in total peripheral resistance index (TPRI), with reductions in renal (RBF), hindquarters (HBF) and mesenteric blood flow (MBF), and leads to the death of all control animals within 30 min. Histamine (100 nmol) injected intracerebroventricularly (i.c.v.) in the early phase of critical hypotension produces a prompt and long-lasting increase in MAP and heart rate, with a 100% survival for 2 h after treatment. The effects are associated with the rise in the circulating blood volume and CI, and the decrease in TPRI, with the increase in RBF and HBF, and persistently lowered MBF. Both splenectomy and ligation of the suprahepatic veins inhibit histamine-induced increase in circulating blood volume as well as cardiac and regional haemodynamic effects. It can be concluded that histamine administered icv activates central endogenous compensatory mechanisms, which leads to the reversal of haemorrhagic shock conditions due to the mobilisation of blood from venous reservoirs, the increase in circulating blood volume and its redistribution. Moreover, histamine evokes the rises in Cl and perfusion of the renal and skeletal muscle vascular regions. PMID- 11939719 TI - Sensory nerves in central and peripheral control of pancreatic integrity by leptin and melatonin. AB - Central nervous system affects pancreatic secretion of enzymes however, the neural modulation of acute pancreatitis has not been investigated. Leptin and melatonin have been recently reported to affect the inflammatory response of various tissues. The identification of specific receptors for both peptides in the pancreas suggests that leptin and melatonin could contribute to the pancreatic protection against inflammation. The aim of this study was: 1/ to compare the effect of intracerebroventricular (i.c.v.) or intraperitoneal (i.p.) administration of leptin or melatonin on the course of caerulein-induced pancreatitis (CIP) in the rat, 2/ to examine the involvement of sensory nerves (SN) and calcitonin gene-related peptide (CGRP) in pancreatic protection afforded by leptin or melatonin, 3/ to assess the effect of tested peptides on lipid peroxidation products (MDA + 4-HNE) in the pancreas of CIP rats, 4/ to investigate the influence of leptin or melatonin on nitric oxide (NO) release from isolated pancreatic acini and 5/ to determine the effects of caerulein and leptin on leptin receptor gene expression in these acini by RT-PCR. CIP was induced by subcutaneous (s.c.) infusion of caerulein (25 microg/kg) to the conscious rats, confirmed by the significant increases of pancreatic weight and plasma amylase and by histological examination. This was accompanied in marked reduction of pancreatic blood flow and significant rise of MDA + 4-HNE in the pancreas. Leptin or melatonin were administered i.p. or i.c.v. 30 min prior to the start of CIP. Deactivation of SN was produced by s.c. capsaicin (100 mg/kg). An antagonist of CGRP, CGRP 8-37 (100 microg/kg i.p.), was given together with leptin or melatonin to the CIP rats. MDA + 4-HNE was measured using LPO commercial kit. NO was determined using the Griess reaction. Pretreatment of CIP rats with i.p. leptin (2 or 10 microg/kg) or melatonin (10 or 50 mg/kg) significantly attenuated the severity of CIP. Similar protective effects were observed following i.c.v. application of leptin (0.4 or 2 microg/rat) but not melatonin (10 or 40 microg/rat) to the CIP rats. Capsaicin deactivation of SN oradministration of CGRP 8-37 abolished above beneficial effects of leptin on CIP, whereas melatonin-induced protection of pancreas was unaffected. Pretreatment with i.p. melatonin (10 or 50 mg/kg), but not leptin, significantly reduced MDA + 4-HNE in the pancreas of CIP rats. Leptin (10(-10) - 10(-6) M) but not melatonin (10(-8) - 10(-5) M) significantly stimulated NO release from isolated pancreatic acini. Leptin receptor gene expression in these acini was significantly increased by caerulein and leptin. We conclude that 1/ central or peripheral pretreatment with leptin protects the pancreas against its damage induced by CIP, whereas melatonin exerts its protective effect only when given i.p., but not following its i.c.v. adminstration, 2/ activation of leptin receptor in the pancreatic acini appears to be involved in the beneficial effects of leptin on acute pancreatitis, 3/ the protective effects of leptin involve sensory nerves, CGRP and increased generation of NO whereas melatonin-induced protection of the pancreas depends mainly on the antioxidant local effect of this indole, and scavenging of the radical oxygen species in the pancreatic tissue. PMID- 11939721 TI - Influence of the corticotropin releasing hormone (CRH) on the brain-blood barrier permeability in cerebral ischemia in rats. AB - The increase in the blood-brain barrier (BBB) permeability and a developing cerebral oedema due to the ischemic infarction appear a few hours, and intensify during a few days, after closing the carotid arteries. It fails to be clear, however, what causes the increase in the microvessels damage, and whether the damage is a secondary result of the vasoactive substances released by the neurones and glia cells damaged by the ischemia. CRH, which plays an essential role in integrative the nervous, endocrine, and immunological systems, has a positive effect on the decrease in the permeability of the BBB damaged by various physical and chemical factors. Therefore, the examination of the CRH role in the cerebral ischemia may prove useful for explaining the processes taking place in the foci of the cerebral infarction and their environment. The experiment was carried out on rats which, 20 minutes before closing of both internal carotid arteries, was administered 10 microg CRH to cerebrospinal fluid via cisterna magna of the brain. The BBB permeability was measured 30 minutes, 3 hours, 3 days, and 7 days after closing the arteries. The experiment has shown the CRH protective effect on the BBB and its consequent effect on the decrease in the BBB permeability which appears in the 3 hours after closing the arteries (p<0.05), and is high significant during the chronic phase of the cerebral ischemia (p<0.03). It can be thus concluded that CRH, by affecting directly the endothelium of the cerebral vessels, decreases the endothelial damage in the acute phase of the ischemia. The decrease is noted to be more significant in the chronic phase of the ischemia; such an effect can be attributed to CRH stimulating the hypothalamic-adrenal axis, and to the secondary activation of the mechanisms decreasing the BBB permeability. PMID- 11939722 TI - Cytokines regulate plasminogen activation system in astrocytoma cells. AB - We report here that human astrocytoma cell line U373-MG is able to express genes of the following components of plasminogen activation system: PA1-1, PN-1, u-PA and t-PA. Treatment of these cells with IL-1beta results in accumulation of PA1 1, PN-1 and u-PA mRNAs, whereas t-PA mRNA remains unaffected. IFNy preferentially enhances PN-1 and PA1-1, EGF enhances PA1-1, u-PA and t-PA expression. Simultaneous addition of anti-inflammatory cytokines IL-4, IL-13 and IL-10 has little effect on the tested components, except induction of u-PA mRNA wich was further enhanced by IL-4. We have confirmed interesting time-dependent regulation of plasminogen activation system by EGF/IFNgamma. Cells stimulated with EGF/IFNgamma show at first increased proteolytic activity but after 24 h inhibition of proteolysis with PA1-1 would prevail. To understand the cooperative effect of EGF and IFNgamma in PA1-1 induction the kinetics of activation of STAT1 was studied. It was found that although EGF alone does not activate STAT1, the STAT1 binding activity in the cells treated with the mixture of EGF/IFNgamma was considerably prolonged. Our results indicate the importance of inflammatory cytokines and EGF in gene regulation of plasminogen activation system in astrocytoma cells. PMID- 11939723 TI - Advocating for patient preference in cervical cytology screening. PMID- 11939724 TI - Toward objective quality assurance: the eyes don't have it. PMID- 11939725 TI - Willingness to pay for new Papanicolaou test technologies. AB - The amount of money a woman is willing to pay for liquid-based cytology technology has not been measured. In the present study, 175 women answered a questionnaire asking how much they would pay to decrease their risk of dying of cervical cancer if a new (liquid-based) Papanicolaou (Pap) test was used in place of the conventional smear. When women assumed that the new Pap test reduced the risk of dying of cervical cancer from 1 in 37,000 to 1 in 50,000, the mean amount they were willing to pay was $237. If women had more than 2 children, they were willing to pay more for the new Pap test than women with 2 or fewer children. These data indicate that liquid-based and conventional Pap tests are undervalued and that cost-effectiveness studies generally have not taken into account the preference of women for new Pap test technologies. PMID- 11939726 TI - Papanicolaou tests diagnosed as atypical by a cytotechnologist and downgraded to benign by a pathologist: a measure of laboratory quality. AB - Follow-up of Papanicolaou (Pap) tests diagnosed as atypical squamous cells of undetermined significance (ASCUS) or atypical glandular cells of undetermined significance (AGUS) by a cytotechnologist and downgraded to benign by a pathologist has not been measured. Squamous intraepithelial lesion (SIL) follow up rates were obtained for Pap tests diagnosed as ASCUS (288) or AGUS (94) and downgraded to benign and for Pap tests diagnosed as repair (231). Statistically significant associations were seen between 7 cytotechnologists and between 7 pathologists and ASCUS, AGUS, downgraded ASCUS, and downgraded AGUS rates. The percentage of downgraded ASCUS cases compared with all ASCUS cases per pathologist ranged from 4.8% to 43.7%. Statistically significant associations between pathologists and SIL follow-up rates for downgraded ASCUS diagnoses were seen. The SIL follow-up rate for repair (7.9%) was similar to that for a downgraded ASCUS (11.0%) or AGUS (7.3%). The parameters of downgraded ASCU and AGUS Pap test interpretations are good quality indicators of individual performance and overall laboratory quality. PMID- 11939727 TI - WT1 immunoreactivity in uterine papillary serous carcinomas is different from ovarian serous carcinomas. AB - WT1 diffusely stains most ovarian serous carcinomas; reactivity of uterine papillary serous carcinomas has not been evaluated. We studied WT1 expression in 13 International Federation of Gynecology and Obstetrics stage 1 and 5 stage 3 or 4 uterine papillary serous carcinomas without ovarian metastases and compared their reactivity with the WT1 staining of 30 ovarian serous carcinomas. WT1 reactivity was evaluated with the C19 and 6F-H2 antibody clones. All 18 uterine papillary serous carcinomas were nonreactive for WT1. The nonovarian metastases of the 5 high-stage uterine papillary serous carcinomas also were nonreactive for WT1. In contrast, 29 (97%) of 30 ovarian serous carcinomas were reactive for WT1. WT1 reactivity in an unknown primary serous carcinoma would suggest it is from a nonuterine site. The mechanisms underlying these findings are unknown. They raise the possibility of genetic differences between the 2 morphologically similar neoplasms. PMID- 11939728 TI - Up-regulation of cyclooxygenase-2 expression in lymphocytic thyroiditis and thyroid tumors: significant correlation with inducible nitric oxide synthase. AB - To cast light on relations of cyclooxygenase-2 (COX-2) expression to lymphocytic thyroiditis and thyroid tumorigenesis, protein levels were immunohistochemically assessed and compared with inducible nitric oxide synthase (iNOS) in a total of 181 cases: follicular adenoma, 23; well-differentiated papillary carcinoma, 85; poorly differentiated papillary carcinoma, 25; anaplastic carcinoma, 7; and follicular carcinoma, 41. In addition, 72 specimens of normal follicular epithelia and 36 of lymphocytic thyroiditis were used as control samples. Immunohistochemical results were confirmed in 2 cases each of normal thyroid, lymphocytic thyroiditis, and well-differentiated and poorly differentiated papillary carcinoma, by Western blotting assay. Stepwise increments in overexpression of COX-2 and iNOS were revealed in epithelial cells of lymphocytic thyroiditis, follicular adenoma, and papillary carcinoma; normal thyroid epithelium showed little expression. A significant positive correlation between the 2 enzymes was found with all cases. Enhanced expression of both COX-2 and iNOS suggests important roles in the inflammatory processes underlying lymphocytic thyroiditis and thyroid tumorigenesis. PMID- 11939729 TI - The morphologic spectrum of metastatic prostatic adenocarcinoma to the lung: special emphasis on histologic features overlapping with other pulmonary neoplasms. AB - We undertook a detailed histologic study to identify specific morphologic features that may aid in distinguishing prostatic adenocarcinoma with lung metastases (PALM) from other pulmonary tumors with similar histologic features. In 16 cases, we found 3 predominant architectural patterns: microacinar (n = 10), tubulopapillary (ductal; n = 4), and carcinoid-like (n = 2). Characteristic features of PALM included small acinar and/or cribriform growth, frequent lymphangitic permeation, lack of stromal response, uniform round nuclei with prominent nucleoli, intraluminal blue mucin, and prominent cell borders. By immunohistochemical staining, prostate-specific antigen and prostate-specific acid phosphatase were present in 13 of 14 and 13 of 13 cases, respectively. Metastatic prostatic duct adenocarcinoma exhibited morphologic features similar to metastatic colonic adenocarcinoma. Two cases had a carcinoid-like appearance with nested or solid architecture, parachromatin clearing, and prominent nucleoli, but lacked the finely stippled chromatin pattern of carcinoid tumors. Several features that may result in misinterpretation or lack of association of the neoplasm in the lung with a prostatic primary include lung metastasis preceding the detection of a prostatic primary tumor, solitary pulmonary nodule, tubulopapillary (ductal) or carcinoid-like pattern, scant material in which histologic features of metastatic prostate carcinoma are not fully appreciated, and frequent necrosis. Attention to specific discriminating histologic features, supported by immunohistochemical staining, may be useful in the differential diagnosis, which is therapeutically and prognostically critical. PMID- 11939730 TI - Intratumoral genetic heterogeneity in Barrett adenocarcinoma. AB - In 10 cases of Barrett adenocarcinoma, samples from 8 tumor areas (including superficial and deep from peripheral and central areas) and a regional lymph node metastasis were studied for amplification of c-myc, c-erbB-2, and EGFR. We analyzed loss of heterozygosity (LOH) at 3 loci (APC, MCC, and RB) and 2 anonymous microsatellite markers (D4S1652 and D18S474). We detected c-myc in variable fractions of tissue samples from 3 of 9 tumors; EGFR was amplified in 2 specimens from 1 tumor. One tumor demonstrated amplification of c-erbB-2 in all areas. LOH at the D4S1652, MCC, RB, APC, and D18S474 loci was found in 75% (3/4), 57% (4/7), 50% (4/8), 11% (1/9), and 0% (0/10) of informative cases, respectively. LOH generally was restricted to variable subpopulations of tumor cells within individual tumors. There was no obvious association of certain genetic alterations with topographically distinct tumor regions; however, superficial areas showed more frequent genetic alterations than areas from the deeply invading front. More aberrations were detected in the periphery than in the center. Barrett adenocarcinoma is characterized by marked intratumoral genetic heterogeneity, which must be considered when evaluating genetic alterations as indicators of response to therapy and prognosis. PMID- 11939731 TI - Cytohistologic discrepancies: a means to improve pathology practice and patient outcomes. AB - The use of cytohistologic discrepancies to investigate and reduce error seldom is studied. All gynecologic discrepancies (n = 283; 0.87% and 7.37% of all cytologic and histologic cases, respectively) and nongynecologic discrepancies (n = 146; 2.26% and 0.44% of all cytologic and histologic cases, respectively) for 26 months were classified as sampling or interpretive. Specimen type and pathologist discrepancy percentages, effect of discrepancies on patient outcome, and interobserver agreement of discrepancies were evaluated. Discrepancies were interpretive in 67% and 34% of gynecologic and nongynecologic cases, respectively. Statistically significant associations were seen between individual pathologist and discrepancy percentages. Breast (1.2%) and bronchial (0.8%) cytologic diagnoses had the highest discrepancy percentages. The kappa scores ranged from 0.02 to 0.45 for pairwise agreement of discrepant cases. Of nongynecologic interpretive discrepancies available for review, 63% (27/43) and 14% (6/43) were of no or minor clinical significance, respectively. Cytohistologic correlation is a useful tool to monitor performance and to identify specimen types prone to error. PMID- 11939732 TI - Primary cutaneous diffuse large B-cell lymphoma: a clinicopathologic study of 15 cases. AB - Primary cutaneous diffuse large B-cell lymphoma (DLBCL) is an uncommon lymphoma. Some authors have suggested that large B-cell lymphoma can be segregated based on anatomic site, with tumors of the lower extremity being unique. We report 15 cases of primary cutaneous DLBCL. Each case was analyzed immunohistochemically using antibodies specific for CD3, CD5, CD10, CD20, bcl-2, bcl-6, and p53. Polymerase chain reaction analysis for t(14;18)(q32;q21) also was performed. There were 13 men and 2 women (median age, 64 years). Thirteen tumors were composed predominantly of centroblasts, and 2 were immunoblastic. There was a median follow-up of 72 months. Of the 4 patients with primary cutaneous DLBCL of the lower extremity (thigh, knee, leg), 2 (50%) experienced a recurrence and 1 patient died of disease. In the non-lower extremity cases, 18% (2/11) recurred and no patients died of disease. We conclude that primary cutaneous DLBCL usually occurs in elderly patients with a male predominance. Recurrences are common, but death of disease is rare. PMID- 11939733 TI - The molecular characterization of fatal infectious mononucleosis. AB - We describe a retrospective study of 4 cases of sporadic fatal infectious mononucleosis (IM), 1 case of fatal IM, and 1 case of sporadic severe IM. Patients were 26 months to 17 years old; 3 were male. Five died of complications of IM. All 5 of these patients had the Epstein-Barr virus (EBV) present in examined tissue specimens; EBV was monoclonal in 3 patients and biclonal in 1. EBV clonality studies were not performed in the remaining patient. All 5 patients also had monoclonal gene rearrangements. The sixth patient survived despite a life-threatening clinical course; EBV was oligoclonal, and gene rearrangements were not detected. EBV clonality and gene rearrangement studies may be usefulfor predicting which patients with clinically aggressive IM are at highest risk for fatal outcome. Patients in whom IM has a fatal outcome are more likely to have monoclonal or biclonal EBV and immunoglobulin heavy chain or T-cell receptor gene rearrangements. In contrast, patients with nonfatal IM may lack monoclonal EBV and monoclonal rearrangements of the aforementioned genes. The reasons EBV induces a monoclonal proliferation only in some patients remain to be elucidated. PMID- 11939734 TI - A multiplexed fluorescent microsphere immunoassay for antibodies to pneumococcal capsular polysaccharides. AB - We developed a multiplexed indirect immunofluorescent assay for antibodies to pneumococcal polysaccharides (PnPs) based on the Luminex multiple analyte profiling system (Luminex, Austin, TX). The assay simultaneously determines serum IgG concentrations to 14 PnPs serotypes: 1, 3, 4, 5, 6B, 7F, 8, 9N, 9V, 12F; 14, 18C, 19F, and 23F. To assess the specificity of the multiplexed assay for each individual serotype, inhibition-of-binding studies were conducted using adult serum samples obtained after pneumococcal vaccination. Except for the closely related serotypes 9V and 9N, we demonstrated inhibition by homologous serotypes of more than 95% and inhibition by heterologous serotypes of less than 15% for all 14 PnPs serotypes. There was, however, high heterologous inhibition of 50% or greater with some serotypes. These cross-reacting antibodies could not be removed by preabsorption with pneumococcal C-polysaccharide but were removed by additional preabsorption with serotype 22F polysaccharide. The multiplexed Luminex assay showed good overall agreement with a well-established enzyme-linked immunosorbent assay that is currently recommended for evaluation of pneumococcal vaccine immunogenicity. PMID- 11939735 TI - Comparison of staining methods and a nested PCR assay to detect Histoplasma capsulatum in tissue sections. AB - To optimize diagnosis of histoplasmosis in tissue sections, 30 spleen specimens from mice, experimentally infected with Histoplasma capsulatum, were examined by H&E, Grocott stain, anti-bacille Calmette-Guerin antibody immunostain, Fungiqual A fluorochrome stain (Drs Reinehr and Rembold, Kandern, Germany), and a nested polymerase chain reaction (PCR) assay. Results were compared with the tissue burden determined by quantitative culture. By applying logistic regression, the nested PCR assay was the most sensitive method, but not significantly more sensitive than the Grocott stain. The 50% quantile to achieve a positive result was determined to be 3 colony-forming units per milligram of spleen tissue for the PCR assay, 11 for the Grocott stain, 27 for the fluorochrome stain, 190 for immunostaining, and 533 for the H&E stain. The Grocott and fluorochrome stains did not differ significantly in detecting fungal elements. The PCR assay unambiguously identified H. capsulatum in tissue sections. PMID- 11939736 TI - Laboratory identification of cryoglobulinemia from automated blood cell counts, fresh blood samples, and blood films. AB - Four cases showing different means to detect cryoglobulins are reported: effects on blood cell counts performed on 2 technologically different automated hematology instruments and microscopic features in fresh blood samples and on May Grunwald-Giemsa-stained bloodfilms. These cases were chosen for their instructive value in depicting all artifacts associated with cryoglobulins. Laboratory recognition of the cryoglobulins is important to correct factitious results with automated blood cell counters, mainly pseudoleukocytosis and pseudothrombocytosis. Moreover cryoglobulin-induced laboratory artifacts may be the first factor prompting the assessment for cryoglobulinemia and the diagnosis of the underlying cause. PMID- 11939737 TI - Immunophenotypic variability of B-cell non-Hodgkin lymphoma: a retrospective study of cases analyzed by flow cytometry. AB - Flow cytometric analysis is important in the diagnosis, classification, and follow-up of non-Hodgkin lymphoma. It is assumed that the lymphoma phenotype for each patient remains unchanged over time and is consistent from one specimen to another. To determine the variability in expression of lymphoid antigens, we reviewed 211 flow cytometry specimens of malignant lymphoma from 81 patients. Some antigens showed a stable pattern of expression such as CD5, CD10, CD19, CD20, and HLA-DR. In contrast, CD21, CD22, CD23, and CD25 showed more variability from one specimen to another. We believe several factors affect the stability of antigen expression. True differences in expression most probably are related to the biology and function of the different antigens. For instance, CD19 and CD20 are essential in cell maturation and function and, therefore, are present on the majority of cells. In contrast, CD22 has a role during B-cell activation and, therefore, is more variable. Lack of standardization inflow cytometry procedures also is responsible for some variability. Instrument settings for adequate compensation and the criteria used to determine when an antigen is reported as positive are important considerations when evaluating flow cytometry histograms. PMID- 11939738 TI - Erythrocyte sedimentation rate: use of fresh blood for quality control. AB - The erythrocyte sedimentation rate (ESR) remains the most widely used laboratory test for monitoring the course of infections, inflammatory diseases, and some types of cancer. Several test methods have been developed recently, and as a result the safety and reliability of ESR testing procedures have improved. The method recommended by the International Council for Standardization in Haematology and the National Committee for Clinical Laboratory Standards for ESR measurement is based on the traditional Westergren method, using EDTA anticoagulated samples without dilution. In clinical laboratories, reliable methods for calibration and the use of appropriate control materials are requiredfor monitoring the accuracy and precision of the routine method. We describe and evaluate a procedure for achieving the daily quality control of ESR and for establishing the limits of agreement between working and reference methods. Data from routine patient samples were used to calculate the daily cumulative mean and to monitor its reproducibility over time. Finally, to monitor analytic performance, a comparison was made between results from the measurement of ESR in specimens stored at 4 degrees C for 24 hours and results obtained in fresh samples. PMID- 11939739 TI - Spontaneous regression of Kikuchi lymphadenopathy with oligoclonal T-cell populations favors a benign immune reaction over a T-cell lymphoma. AB - To aid in the initial diagnosis of Kikuchi lymphadenitis and to assess whether the composition of the T cells might shed light on the pathogenesis, we used nested polymerase chain reaction tests followed by high-resolution gel electrophoresis to determine the pattern of T-cell antigen receptor rearrangement in 56 consecutive cases. Except for 1 unusual case with recurrent lymphadenopathy, none had a monoclonal beta or gamma rearrangement. Eight cases had a polyclonal pattern at both beta and gamma loci, 20 cases had a mixed polyclonal beta and oligoclonal gamma pattern, and 27 cases had an oligoclonal pattern at both loci. The high frequency of oligoclonality did not indicate an early-stage T-cell lymphoma in evolution, as confirmed by spontaneous resolution of the lymphadenopathy in all cases within 6 months. Rather, it is consistent with reports of oligoclonal T cells in a variety of immune reactions. We conclude that, in the vast majority of cases, absence of a monoclonal T-cell receptor rearrangement excludes the possibility of T-cell lymphoma, and the presence of an oligoclonal pattern implies a benign immune reaction. PMID- 11939740 TI - The usefulness of immunohistochemistry in the diagnosis of follicular lymphoma in bone marrow biopsy specimens. AB - We used a panel of paraffin antibodies to determine whether neoplastic and nonneoplastic lymphoid aggregates in the bone marrow can be distinguished reliably. Formalin-fixed, paraffin-embedded bone marrow core biopsy specimens with lymphoid aggregates were stained using primary antibodies directed against bcl-2, bcl-6, CD5, CD10, CD20, and CD23. We studied 61 cases (26 follicular lymphoma and 35 benign or atypical aggregates). We found that no single stain is sufficient for identification of neoplastic lymphoid aggregates. However, this distinction was made possible by using a panel of antibodies. Under the conditions we tested, the most useful antibodies were CD10, bcl-2, CD5, and CD20. Most benign or atypical aggregates do not express CD10 and CD23. In addition, nonneoplastic aggregates had a large population of T cells. bcl-2 was useful in an architectural context for distinguishing neoplastic aggregates. bcl-6 often was expressed in both neoplastic and nonneoplastic aggregates and, thus, poorly discriminated between these processes. We studied the expression of CD10 and bcl 6 in selected lymph nodes in some cases. PMID- 11939741 TI - Discrepancies in the immunophenotype of lymphoma cells in samples obtained simultaneously from different anatomic sites. AB - Few studies have compared the immunophenotypic profiles of non-Hodgkin lymphoma (NHL) cells obtained simultaneously from different anatomic sites. In the present study, we compared flow cytometry immunophenotypic results in 64 consecutive NHL cases in which aspiration or biopsy of 2 sites was performed within 30 days to assess the potential discrepancy rate. In 14 cases (22%), discordant antigen expression was identified, including 4 (36%) of 11 cases with discordant morphologic features and 10 (19%) of 53 cases with concordant morphologic features in the 2 samples. Discrepancies involved 1 antigen in 10 patients and 2 antigens in 4 patients. Antigens most frequently discrepant included CD5 (n = 4), FMC7 (n = 3), and CD20 (n = 3). We conclude that the immunophenotype of NHL cells is generally stable, yet discrepancies can occur in a subset of patients. Differences in immunophenotype may relate to mechanisms of disease dissemination, influence of the microenvironment, or differential response to therapy. PMID- 11939742 TI - Leukemias resembling acute promyelocytic leukemia, microgranular variant. AB - Acute promyelocytic leukemia (APL) should be distinguished from other subtypes of acute myeloid leukemia (AML) because of the increased risk of disseminated intravascular coagulation (DIC) and its response to arsenic compounds and retinoids. Some cases of AML seem morphologically similar to the microgranular variant of APL (French-American-British [FAB] AML-M3v) but lack the t(15;17). We evaluated 8 cases of APL-like leukemias for subtle morphologic, cytochemical, immunophenotypic, and cytogenetic differences compared with 5 cases of promyelocytic leukemia/retinoic receptor alpha (PML/RARalpha)-positive APL (FAB AML-M3v). We also evaluated both groups for the presence of DIC. No differences among the groups were noted in blast size, chromatin pattern, nuclear morphologic features, intensity of myeloperoxidase staining, or presence of Auer rods. Immunophenotypes were similar; both types of cases lacked CD34 and HLA-DR and were CD13+ and CD33+. Two cases of APL-like leukemias also were CD56+. DIC was present in 2 patients with M3v. Our study shows that there are no definitive morphologic, cytochemical, or immunophenotypic findings that can distinguish these cases from PML/RARalpha-positive APL. PMID- 11939743 TI - CD10 expression and survival. PMID- 11939744 TI - Intravascular lymphomatosis in T-hepatosplenic lymphoma. PMID- 11939745 TI - Grading systems for DCIS. PMID- 11939746 TI - Implementing best practice models by using a consensus-building process. AB - The Family Education Project is an example of how the consensus-building model can be used to gain acceptance of evidence-based practice, and adapt and implement it on the local level. The authors describe the consensus-building process and the lessons learned from implementing family education in one community in order to show how this process may be used to adapt best practice models to meet unique needs within local communities. PMID- 11939747 TI - Correlates of family burden under medicaid managed mental health care. AB - This study examined predictors of family burden (assistance in daily living, supervision, and subjective concern) for family members of Medicaid recipients with severe mental illness in two regions of Virginia. In the Richmond area, mental health services were provided on a no-risk fee-for-service basis, while in Tidewater these services were provided through a risk-based capitated contract with a managed care organization. No differences in family burden were attributable to the risk-based payment system. Predictors of increased family burden were (a) more reported client symptoms and disruptive behaviors, (b) status as a parent, and (c) living with the client. PMID- 11939748 TI - The impact of realignment on utilization and cost of community-based mental health services in California. AB - Decentralization of California's public mental health system under program realignment has changed the utilization and cost of community-based mental health services. This study examined a sample of 75,951 users, representing 1.5 million adults who visited California's public mental health services during a 6-year period (FY 1988-1990 and FY 1992-1994). Regression analysis was performed to examine cost and utilization reduction over time, across regions, and across psychiatric diagnoses. Overall utilization and cost of community-based mental health services dropped significantly after the implementation of realignment. They were significantly lower for (a) 24-hour services in the urban industrialized Southern Region and (b) outpatient services in the agricultural Central Region of the state. Users diagnosed with mood disorders took a greater portion, but were associated with significantly less treatment and cost than other users in the post-realignment period. When local communities bear the financial risks and rewards, they find more efficient methods of delivering community-based mental health services. PMID- 11939749 TI - Consumer satisfaction and incarceration after treatment. AB - Consumer satisfaction is increasingly used to measure community mental health program performance. Understanding the relationship between consumer satisfaction and treatment outcomes is only beginning. This article adds to this understanding by reporting on an assessment of the relationship between consumer evaluation of community mental health services and incarceration after treatment in a statewide system of care. Results indicate that satisfaction with services is related to incarceration after treatment, with satisfied consumers having lower incarceration rates. These results support the use of self-reported consumer satisfaction as a measure of mental health program performance. PMID- 11939750 TI - Deficiency citations for mental health care in nursing homes. AB - The Centers for Medicare and Medicaid Services (CMS) certifies approximately 17,000 nursing homes per year. When a facility does not meet a standard, a deficiency citation is issued. Using 1998 citations, we examined the structure and process characteristics of nursing homes that were associated with mental health deficiencies. Our findings showed that (a) some nursing homes have a high number of deficiencies in the area of mental health care; (b) these deficiencies do not appear to be associated with specific processes used in nursing homes; and (c) some structural factors such as staffing, private-pay occupancy, and Medicare occupancy appear to have an important influence on the number of mental health care deficiencies. PMID- 11939751 TI - Community vs. private-practice psychiatrists: lifestyle and role as related to career track. PMID- 11939752 TI - Managed mental health care in medicaid: does the solution match the problem? PMID- 11939753 TI - Evaluation of innovative skin-marking technique performed before thyroid ultrasound-guided fine-needle aspiration biopsies. AB - OBJECTIVE: To introduce an innovative skin-marking technique performed before ultrasound-guided fine-needle aspiration biopsy (US-FNAB) of the thyroid. METHODS: We studied 248 patients with thyroid nodules, who were classified on the basis of physical examination into two groups-those with palpable nodules (N = 127) and those with nonpalpable or difficult-to-palpate nodules (N = 121). Each group was further subdivided according to the size of the thyroid nodule (< or = 15 mm versus >15 mm). Before US-FNAB, we performed the skin-marking technique with the aid of a catheter, a permanent marker, and ultrasound guidance. An established point for needle entry was indicated on the skin. The chi-square test was used to compare results between the groups of patients. RESULTS: The proportions of adequate and insufficient biopsy material in the overall group of patients were 88.7% and 11.3%, respectively. When the patients were stratified by palpable and nonpalpable thyroid nodules, biopsy specimens were adequate in 89.8% and insufficient in 10.2% of those with palpable nodules, whereas the corresponding proportions for those with nonpalpable nodules were 87.6% and 12.4%, respectively. No statistically significant differences were noted between the two groups. Comparisons between patients with thyroid nodules >15 mm in their largest diameter versus those with nodules < or =15 mm also showed no statistically significant differences in terms of insufficient biopsy material. CONCLUSION: Our technique creates a marking on the skin that leads directly to the thyroid nodule and facilitates the acquisition of adequate cytologic material. This is particularly relevant when small transducers are not available or when the physician needs reassurance about the accuracy of the thyroid biopsy site. PMID- 11939754 TI - Description and evaluation of a glycemic management protocol for patients with diabetes undergoing heart surgery. AB - OBJECTIVE: To evaluate a protocol for glycemic management in the treatment of postoperative heart patients with diabetes in the setting of a community hospital. METHODS: The protocol included a standardized guideline in tabular form for nurse-implemented insulin infusion ("drip") therapy for postoperative glycemic control. At the time of discontinuation of the insulin drip, the glycemic status of patients with diabetes was managed by the endocrine department. Overall, 29 patients were assessed without and 29 patients with use of the protocol in a community hospital. RESULTS: From postoperative days 0 through 4, use of the protocol resulted in a greater number of blood glucose determinations, a trend toward greater utilization of insulin drip therapy without a significant increase in the number of patients treated with insulin drip, and no change in the frequency of hypoglycemic episodes. During the same time interval, the percentages of postoperative days during which at least one blood glucose value equaled or exceeded 250 mg/dL were 27.5% without the protocol and 16.8% with use of the protocol (P = 0.0318). The principal finding of the study was reduction in the percentage of postoperative days during which mean blood glucose values equaled or exceeded 200 mg/dL to less than half the previously observed frequency-from 38.4% without the protocol to 16.8% with the protocol (P = 0.0001). The effectiveness of the insulin drip component of the protocol is suggested by a trend, shown on postoperative days 2 through 4, of 70 patient days with mean blood glucose levels <200 mg/dL (58 of these days without insulin drip therapy) and 15 patient days with mean blood glucose values > or =200 mg/dL (none of these days associated with same-day insulin drip therapy). CONCLUSION: A standardized approach to insulin drip therapy, in combination with subspecialty consultation for follow-up glycemic management with use of subcutaneous administration of insulin, can improve glycemic control in postoperative heart patients without continuation of insulin drip therapy outside the critical-care unit. The trends observed on postoperative days 2 through 4, that most patients maintained glycemic control without insulin drip therapy and that all failures of glycemic control occurred among patients who no longer received insulin drip therapy, suggest the possibility of developing criteria for selection of patients for continuation of insulin infusion outside the critical care unit. PMID- 11939755 TI - Early onset of medullary thyroid carcinoma in a kindred with multiple endocrine neoplasia type iia associated with cutaneous lichen amyloidosis. AB - OBJECTIVE: To describe the molecular characterization of a kindred affected by the rare variant of multiple endocrine neoplasia type IIA (MEN IIA) associated with cutaneous lichen amyloidosis and to discuss the clinical implications in the management of this syndrome. METHODS: A kindred with four affected family members was identified, and DNA analysis was performed by sequencing exon 11 of the RET proto-oncogene. Presymptomatic genetic screening was offered to all first-degree relatives. RESULTS: Sequencing analysis of the RET proto-oncogene revealed a Cys634Trp (TGC->TGG) mutation in all clinically affected family members and in an asymptomatic 5-year-old child who, after thyroidectomy, was found to have multicentric medullary thyroid carcinoma and C-cell hyperplasia. A Gly691Ser (GGT >AGT) polymorphism was also detected in this family but did not segregate with the disease. CONCLUSION: To our knowledge, this is the earliest detection of medullary thyroid carcinoma reported thus far in a kindred with MEN IIA associated with cutaneous lichen amyloidosis, and this finding suggests that prophylactic thyroidectomy, in kindreds with this variant, should be performed before the age of 5 years. PMID- 11939756 TI - Thyroid size and iodine intake in iodine-repleted pregnant women in Isfahan, Iran. AB - OBJECTIVE: To evaluate the goiter and iodine intake status of pregnant women in Isfahan, after 8 years of iodized salt distribution in Iran. METHODS: Thyroid staging was assessed by clinical examination, thyroid volume was determined by sonography, and urinary iodine (UI) excretion was assessed by the digestion method in 90 healthy pregnant women (30 in each trimester) and 90 age-matched nonpregnant women selected by random sampling in prenatal and primary health-care clinics. The data were reported as mean +/- standard deviation; P values <0.05 were considered statistically significant. RESULTS: The mean age of the pregnant and the nonpregnant women was 25.3 and 27.5 years, respectively-no significant difference (P = NS). The clinical goiter prevalence in the pregnant and the nonpregnant groups was 37% and 32%, respectively (P = NS). The mean thyroid volume in the pregnant and nonpregnant women was 7.8 +/- 3.2 and 7.8 +/- 2.8 mL, respectively (P = NS). Urinary iodine (UI) excretion was 20.7 +/- 6.9 mg/dL in pregnant women and 23.7 +/- 7.6 mg/dL in nonpregnant women (P = NS). The prevalence of goiter assessed by sonography was 29% in pregnant women and 21% in nonpregnant women (P = NS). The mean thyroid size in 26 of 90 pregnant women with goiter (thyroid volume >9.2 mL) was 11.8 +/- 2.73 mL and in 19 of 90 nonpregnant women with goiter was 12.36 +/- 1.6 mL (P = NS). The mean thyroid volume was 6.0 +/- 1.7, 9.9 +/- 1.7, 11.8 +/- 2.2, and 18.9 +/- 2.4 mL in the pregnant women with or without goiter at thyroid stages 0, Ia, Ib, and II, respectively. A strong correlation between goiter staging assessed by clinical examination and thyroid volume determined by sonography was found in pregnant (r = 0.77) and nonpregnant (r = 0.78) women (both P<0.000001). Mean UI excretion was 20.9 +/- 7.0, 19.9 +/- 6.8, 20.6 +/- 7.5, and 25.9 +/- 2.3 mg/dL in the pregnant women at thyroid stages 0, Ia, Ib, and II, respectively. In the pregnant and the nonpregnant women, no correlation was found between thyroid stage and UI excretion or between thyroid volume and UI excretion. CONCLUSION: No iodine deficiency was found in Isfahani pregnant women. Thus, as in most iodine sufficient areas, thyroid size did not increase during pregnancy. Despite sufficient iodine intake, a moderate prevalence of goiter was noted in pregnant and nonpregnant women. This study also revealed that careful physical examination of the thyroid had diagnostic accuracy similar to sonography. PMID- 11939757 TI - Differentiated thyroid carcinoma with functional autonomy. AB - OBJECTIVE: To present a case of papillary carcinoma in an autonomously hyperfunctioning thyroid nodule. METHODS: We chronicle the clinical and laboratory findings in a patient with a painless neck mass, with a particular focus on the pathologic findings after surgical removal of the right thyroid lobe. RESULTS: A 39-year-old woman had an enlarging nodule of the right thyroid lobe. Results of thyroid function tests suggested subclinical hyperthyroidism. Two months later, the patient complained of increasing swelling in the neck (but still had no symptoms suggestive of hyperthyroidism). Thus, resection of the right thyroid lobe was performed. Pathologic analysis disclosed low-grade papillary thyroid carcinoma within the nodule, with a small rim of compressed inactive-appearing thyroid tissue surrounding the nodule. Subsequently, she underwent total thyroidectomy and follow-up care for thyroid carcinoma. CONCLUSION: Although solitary hyperfunctioning nodules of the thyroid gland are usually considered benign, the current case suggests that the diagnosis of autonomous thyroid nodules does not preclude thyroid carcinoma in a functioning nodule. PMID- 11939759 TI - A fatal case of gastric mucormycosis and diabetic ketoacidosis. AB - OBJECTIVE: To describe a case of gastric mucormycosis in conjunction with severe bleeding in a young woman, which occurred after an episode of ketoacidosis and had a fatal outcome. METHODS: We present a case report, including detailed clinical and pathologic findings in a woman with gastric mucormycosis associated with severe bleeding. RESULTS: A 17-year-old woman sought medical assistance for diabetic ketoacidosis and severe epigastric pain. Chest radiography showed normal findings, and blood and urine cultures were negative for bacterial growth. Endoscopy disclosed an extensive ulcerated lesion involving the greater curvature and posterior wall of the stomach. Biopsy specimens demonstrated the presence of invasive mucormycosis. Treatment with amphotericin B was initiated, but severe persistent gastrointestinal bleeding resulted in the patient's demise. CONCLUSION: In conjunction with diabetic ketoacidosis, severe infection by Mucor has been reported mainly in the rhino-orbital area. Although uncommon, the gastrointestinal tract can also be involved. In the case described, severe hemorrhage ensued and caused the patient's death. PMID- 11939758 TI - Minimal change nephropathy and graves' disease: report of a case and review of the literature. AB - OBJECTIVE: To describe a possible association between Graves' disease and nephrotic syndrome attributable to minimal change nephropathy and to review the literature related to renal diseases in patients with Graves' disease. METHODS: The clinical, laboratory, and renal biopsy findings in a patient with Graves' disease and minimal change nephropathy are discussed. In addition, the pertinent English-language literature published from 1966 to 2001, determined by means of a MEDLINE search, is reviewed. RESULTS: A 29-year-old woman underwent assessment by her primary-care physician because of palpitations, sweating, and a 4.5-kg weight loss. Physical examination revealed a diffuse goiter and tremors of the extremities but no ophthalmologic signs. Laboratory tests confirmed a diagnosis of thyrotoxicosis. Treatment was initiated with propylthiouracil and propranolol. Four weeks later, she presented to the University of Louisville Hospital with increasing swelling of her legs and periorbital puffiness. Examination revealed generalized edema, ascites, and pleural effusion. She continued to have features of thyrotoxicosis. Laboratory tests showed undetectable thyroid-stimulating hormone (<0.03 mIU/mL) and homogeneously increased 123I thyroid uptake and scan. A 24-hour urine collection revealed urinary protein excretion of 6.75 g. Antinuclear antibodies, serum complement levels, hepatitis, and human immunodeficiency virus (HIV) screen were normal. A kidney biopsy specimen revealed features consistent with minimal change disease on light, immunofluorescence, and electron microscopy. The patient had an excellent clinical and laboratory response to treatment with radioactive iodine and corticosteroids, and she was asymptomatic at 6-month follow-up. CONCLUSION: To the best of our knowledge, this is the first report of the concomitant occurrence of Graves' disease and minimal change disease in the absence of any other immunologic disorder known to be associated with minimal change nephropathy. PMID- 11939760 TI - Chronic urticaria: an unusual initial manifestation of primary hyperparathyroidism. AB - OBJECTIVE: To discuss the rare association between primary hyperparathyroidism and chronic urticaria. METHODS: A case report with clinical and laboratory details is presented, and the pertinent literature is reviewed. RESULTS: A 29 year-old patient with chronic urticaria for more than 2 years, which was partially responsive to medical treatment, was found to have primary hyperparathyroidism. After parathyroidectomy, the urticaria resolved, and it has not recurred during a 1-year follow-up period. CONCLUSION: Primary hyperparathyroidism can be a rare cause of chronic urticaria. PMID- 11939761 TI - Metastatic cervical carcinoma with ectopic calcitonin production presenting as a thyroid mass. AB - OBJECTIVE: To describe a case of cervical carcinoma metastatic to the thyroid in association with ectopic production of calcitonin. METHODS: We present the medical history, clinical findings, and laboratory results in a 38-year-old woman with a neck mass, and we discuss the frequency of metastatic involvement of the thyroid. RESULTS: A 38-year-old woman was referred for evaluation of an enlarging neck mass. She had undergone radical hysterectomy and irradiation for invasive cervical carcinoma 1 year earlier. Laboratory investigation showed only mild anemia, high plasma bicarbonate concentration, increased alkaline phosphatase, and mild hypoalbuminemia. Computed tomography of the abdomen and pelvis disclosed multiple lesions in the liver. Fine-needle aspiration biopsy of the thyroid mass revealed a poorly differentiated carcinoma, which stained negative for thyroglobulin but positive for several neuroendocrine tumor markers. Measurement of serum calcitonin showed a dramatically increased level (5,000 pg/mL). The same histologic and immunochemical profile was found in a liver biopsy specimen and in the original cervical tumor. Metastatic neuroendocrine cervical carcinoma was diagnosed, and the patient died 6 months later despite four cycles of chemotherapy. CONCLUSION: To the best of our knowledge, this is the first case report of a neuroendocrine cervical carcinoma manifesting as a palpable thyroid mass, associated with ectopic production of calcitonin. PMID- 11939762 TI - Acromegaly and pheochromocytoma: report of a rare coexistence. AB - OBJECTIVE: To describe a patient with the rare coexistence of acromegaly and pheochromocytoma. METHODS: We report a case of a 57-year-old woman, who was initially examined because of polyarthritis, she was also diagnosed with type 2 diabetes mellitus and hypertension at age 56 years. Her history, clinical findings, laboratory results, and management are summarized, and etiologic hypotheses are discussed. RESULTS: The patient had recurrent headaches and reported an increasing size of her shoes and gloves during the previous 4 years. Enlargement of her hands and feet and a bilateral temporal field defect were noted on examination. Laboratory studies revealed high levels of insulin-like growth factor I (IGF-I) and growth hormone (GH). Magnetic resonance imaging (MRI) showed a 3-cm sellar mass with impingement on the optic chiasm. The plasma level of growth hormone-releasing hormone (GHRH) was normal. She underwent transsphenoidal adenomectomy. Histologic examination confirmed a pituitary adenoma, immunoreactive for GH. Postoperatively, her headaches and arthritic pain diminished, and her levels of IGF-I and GH normalized; however, labile hypertension persisted. The urinary metanephrines and plasma catecholamines were increased. A 3-cm left adrenal mass, seen on abdominal MRI, was removed laparoscopically, after which urinary metanephrines normalized and both the diabetes and the hypertension resolved. Histopathologic analysis confirmed the diagnosis of pheochromocytoma. Immunohistochemical staining was negative for GHRH. CONCLUSION: The finding of a pheochromocytoma and acromegaly could be a fortuitous coexistence of two separate endocrine tumors; however, the probability of such an event is extremely low. A cause-and-effect relationship has been suggested because of previous reports of GHRH production by pheochromocytomas. Some investigators have also suggested that this coexistence might be a multiple endocrine neoplasia variant. Our patient had no evidence of GHRH production, nor did we document any familial autosomal dominant transmission pattern. PMID- 11939764 TI - A new paradigm for control of diabetes. PMID- 11939763 TI - United Kingdom prospective diabetes study: a different perspective. PMID- 11939765 TI - Klinefelter's syndrome manifesting as an acute pulmonary embolus in a 52-year-old man. PMID- 11939766 TI - Ultrasound-guided fine-needle aspiration of thyroid nodules. PMID- 11939767 TI - Insulin infusions for cardiac surgery patients with diabetes: a call to reason. PMID- 11939768 TI - When "The 7-Year Itch" is indicative of an endocrine malignant condition. PMID- 11939769 TI - Mechanism of glutamate receptor-channel function in rat hippocampal neurons investigated using the laser-pulse photolysis (LaPP) technique. AB - Ionotropic glutamate receptors are members of a large family of plasma membrane proteins expressed by cells of the nervous system. Upon binding glutamate, the receptors transiently open transmembrane channels that allow the entry of sodium ions. The resulting changes in the transmembrane potential of the cell initiates a process that is involved in signal transmission to another cell. The binding of glutamic acid triggers the channel opening in the microsecond time domain and the reversible inactivation (desensitization) of the receptors in the millisecond time region. The channel-opening mechanism of glutamate receptors was investigated in rat hippocampal neurons voltage-clamped to -60 mV at room temperature and pH 7.4. Two rapid chemical reaction techniques were used: (1) a cell-flow method with a 4-10 ms time resolution to apply L-glutamate and (2) a laser-pulse photolysis technique to release glutamate from gamma-O-(alpha-carboxy 2-nitrobenzyl)glutamate (alphaCNB-caged L-glutamate) with a time constant of 30 micros. The rate and equilibrium constants for channel opening were determined. The results are consistent with the receptor binding two molecules of glutamic acid before the channel opens, with an apparent dissociation constant of 600 microM. Channel opening and closing rate constants, k(op) and k(cl), were determined to be (9.5 +/- 1) x 10(3) s(-1) and (1.1 +/- 0.1) x 10(3) s(-1), respectively. The value of the channel-opening equilibrium constant, Phi (=k(op)/k(cl)), was 8.6 when determined by laser-pulse photolysis and 6.6 in cell flow experiments. The results suggest that there are at least two forms of glutamate receptors in rat hippocampal neurons that desensitize with different rates. At a concentration of 500 microM glutamate, 80% of the receptors desensitized with a rate of approximately 200 s(-1) and 20% with a rate of approximately 50 s(-1). PMID- 11939770 TI - Identification of a novel archaebacterial thioredoxin: determination of function through structure. AB - As part of a high-throughput, structural proteomic project we have used NMR spectroscopy to determine the solution structure and ascertain the function of a previously unknown, conserved protein (MtH895) from the thermophilic archeon Methanobacterium thermoautotrophicum. Our findings indicate that MtH895 contains a central four-stranded beta-sheet core surrounded by two helices on one side and a third on the other. It has an overall fold superficially similar to that of a glutaredoxin. However, detailed analysis of its three-dimensional structure along with molecular docking simulations of its interaction with T7 DNA polymerase (a thioredoxin-specific substrate) and comparisons with other known members of the thioredoxin/glutaredoxin family of proteins strongly suggest that MtH895 is more akin to a thioredoxin. Furthermore, measurement of the pK(a) values of its active site thiols along with direct measurements of the thioredoxin/glutaredoxin activity has confirmed that MtH895 is, indeed, a thioredoxin and exhibits no glutaredoxin activity. We have also identified a group of previously unknown proteins from several other archaebacteria that have significant (34-44%) sequence identity with MtH895. These proteins have unusual active site -CXXC- motifs not found in any known thioredoxin or glutaredoxin. On the basis of the results presented here, we predict that these small proteins are all members of a new class of truncated thioredoxins. PMID- 11939771 TI - Manganese substantially alters the dynamics of translesion DNA synthesis. AB - The effect of metal ion substitution on the dynamics of translesion DNA synthesis catalyzed by the bacteriophage T4 DNA polymerase was quantitatively evaluated through steady-state and transient kinetic techniques. Substitution of Mn(2+) for Mg(2+) enhances the steady-state rate of dNMP misinsertion opposite an abasic site by 11-34-fold. At the molecular level, the enhancement in translesion DNA synthesis reflects a substantial increase in the rate of the conformational change preceding phosphoryl transfer for all dNTPs that were tested. This is best illustrated by the biphasic pre-steady-state time course of dAMP insertion opposite an abasic site which indicates that a step after chemistry is rate limiting for steady-state enzyme turnover. Furthermore, the k(pol) value of 40 s( 1) measured under single-turnover reaction conditions is 20-fold greater than the k(cat) value of 2 s(-1) measured for steady-state enzyme turnover. Finally, the low elemental effect ( approximately 2.4-fold reduction in k(pol)) measured by substituting the alpha-thiotriphosphate analogue for dATP further argues that chemistry is not rate-limiting. In contrast to the biphasic insertion of dAMP, pre-steady-state time courses for the insertion of dCMP, dGMP, or dTMP opposite an abasic site were linear. Nearly identical k(pol) values ( approximately 1 s( 1)) were measured for the insertion of dCMP, dGMP, and dTMP opposite the abasic site using single-turnover conditions. However, the large elemental effects of 27 and 70 measured by substituting the alpha-thiotriphosphate analogues for dCTP and dGTP, respectively, suggest that phosphoryl transfer may be the rate-limiting step for their insertion opposite the abasic site. Various models are discussed in an attempt to explain the effect of metal ion substitution on the dynamics of translesion DNA replication. PMID- 11939772 TI - Importance of lysine 125 for heparin binding and activation of antithrombin. AB - The anticoagulant sulfated polysaccharide, heparin, binds to the plasma coagulation proteinase inhibitor, antithrombin, and activates it by a conformational change that results in a greatly increased rate of inhibition of target proteinases. Lys125 of antithrombin has previously been implicated in this binding by chemical modification and site-directed mutagenesis and by the crystal structure of a complex between antithrombin and a pentasaccharide constituting the antithrombin-binding region of heparin. Replacement of Lys125 with Met or Gln in this work reduced the affinity of antithrombin for full-length heparin or the pentasaccharide by 150-600-fold at I = 0.15, corresponding to a loss of 25-33% of the total binding energy. The affinity decrease was due both to disruption of approximately three ionic interactions, indicating that Lys125 and two other basic residues of antithrombin act cooperatively in binding to heparin, and to weakened nonionic interactions. The mutations caused a 10-17-fold decrease in the affinity of the initial, weak binding step of the two-step mechanism of heparin binding to antithrombin. They also increased the reverse rate constant of the second, conformational change step by 10-50-fold. Lys125 is thus a major heparin binding residue of antithrombin, contributing an amount of binding energy comparable to that of Arg129, but less energy than Lys114. It is the first residue identified so far that has a critical role in the initial recognition of heparin by antithrombin, but also appreciably stabilizes the heparin-induced activated state of the inhibitor. These effects are exerted by interactions of Lys125 with the nonreducing end of the heparin pentasaccharide. PMID- 11939773 TI - Identification of the active site of gelatinase B as the structural element sufficient for converting a protein to a metalloprotease. AB - Gelatinase B is a member of the matrix metalloproteinase family that efficiently cleaves gelatin, elastin, and types V and X collagen. To understand the contribution of the active site of the enzyme (amino acid residues 373-456) in these activities, we studied catalytic properties of a fusion protein consisting of maltose binding protein and the active site region of gelatinase B. We found that addition of the active site of gelatinase B, which corresponds to 12% of the total protein molecule, to maltose binding protein is sufficient to endow the protein with the ability to cleave the peptide substrates Mca-PLGL(Dpa)AR-NH(2) and DNP-PLGLWA-(D)-R-NH(2). The fusion protein hydrolyzed the Mca-PLGL(Dpa)AR NH(2) peptide with the same efficiency as that of the stromelysin, k(cat)/K(m) approximately 1.07 x 10(6) M(-)(1) h(-)(1). The fusion protein, however, was not able to degrade the large substrate, gelatin. Inhibition of the activity of the protein by EDTA suggested that its activity was metal dependent. ESR analyses indicated that the fusion protein bound one molecule of Zn(2+). In addition, Z Pro-Leu-Gly-hydroxamate and TIMP-1 inhibited the activity of the protein, suggesting that the structure of the active site of the fusion protein is similar to that of the other metalloproteinases. These data provide fundamental information about the structural elements required for transforming a protein to a metalloprotease. PMID- 11939774 TI - Three-dimensional structure of the L-threonine-O-3-phosphate decarboxylase (CobD) enzyme from Salmonella enterica. AB - The three-dimensional structure of the pyridoxal 5'-phosphate (PLP)-dependent L threonine-O-3-phosphate decarboxylase (CobD) from Salmonella enterica is described here. This enzyme is responsible for synthesizing (R)-1-amino-2 propanol phosphate which is the precursor for the linkage between the nucleotide loop and the corrin ring in cobalamin. The molecule is a molecular dimer where each subunit consists of a large and small domain. Overall the protein is very similar to the members of the family of aspartate aminotransferases. Indeed, the arrangement of the ligands surrounding the cofactor and putative substrate binding site are remarkably close to that observed in histidinol phosphate aminotransferase, which suggests that this latter enzyme might have been its progenitor. The only significant differences in structure occur at the N terminus, which is approximately 12 residues shorter in CobD and does not form the same type of interdomain interaction common to other aminotransferases. CobD is unusual since within the aspartate aminotransferase subfamily of PLP-dependent enzymes the chemical transformations are substantially conserved, where the only exceptions are 1-aminocyclopropane-1-carboxylate synthase and CobD. Although there are a large number of PLP-dependent amino acid decarboxylases, these are generally larger and structurally distinct from the members of the aspartate aminotransferase subfamily of enzymes. The structure of CobD suggests that the chemical fate of the external aldimine can be redirected by modifications at the N-terminus of the protein. This study provides insight into the evolutionary history of the cobalamin biosynthetic pathway and raises the question of why most PLP-dependent decarboxylases are considerably larger enzymes. PMID- 11939775 TI - Unraveling the substrate-metal binding site of ferrochelatase: an X-ray absorption spectroscopic study. AB - Ferrochelatase (EC 4.99.1.1), the terminal enzyme of the heme biosynthetic pathway, catalyzes the insertion of ferrous iron into the protoporphyrin IX ring. Ferrochelatases can be arbitrarily divided into two broad categories: those with and those without a [2Fe-2S] center. In this work we have used X-ray absorption spectroscopy to investigate the metal ion binding sites of murine and Saccharomyces cerevisiae (yeast) ferrochelatases, which are representatives of the former and latter categories, respectively. Co(2+) and Zn(2+) complexes of both enzymes were studied, but the Fe(2+) complex was only studied for yeast ferrochelatase because the [2Fe-2S] center of the murine enzyme interferes with the analysis. Co(2+) and Zn(2+) binding to site-directed mutants of the murine enzyme were also studied, in which the highly conserved and potentially metal coordinating residues H207 and Y220 were substituted by residues that should not coordinate metal (i.e., H207N, H207A, and Y220F). Our experiments indicate four coordinate zinc with Zn(N/O)(3)(S/Cl)(1) coordination for the yeast and Zn(N/O)(2)(S/Cl)(2) coordination for the wild-type murine enzyme. In contrast to zinc, a six-coordinate site for Co(2+) coordinated with oxygen or nitrogen was present in both the yeast and murine (wild-type and mutated) enzymes, with evidence of two histidine ligands in both. Like Co(2+), Fe(2+) bound to yeast ferrochelatase was coordinated by approximately six oxygen or nitrogen ligands, again with evidence of two histidine ligands. For the murine enzyme, mutation of both H207 and Y220 significantly changed the spectra, indicating a likely role for these residues in metal ion substrate binding. This is in marked disagreement with the conclusions from X-ray crystallographic studies of the human enzyme, and possible reasons for this are discussed. PMID- 11939776 TI - A comparative resonance Raman analysis of heme-binding PAS domains: heme iron coordination structures of the BjFixL, AxPDEA1, EcDos, and MtDos proteins. AB - The heme-PAS is a specialized domain with which a broad class of signal transducing heme proteins detect physiological heme ligands. Such domains exhibit a wide range of ligand binding parameters, yet they are all expected to feature an alpha-beta heme binding fold and a predominantly hydrophobic heme distal pocket without a distal histidine. We have compared, for the first time, the resonance Raman spectra of several heme-PASs: the heme-binding domains of Bradyrhizobium japonicum FixL, Escherichia coli Dos, Acetobacter xylinum PDEA1, and Methanobacterium thermoautotrophicum Dos. In all cases, the nu(Fe)-(CO) and nu(C-O) values of the carbonmonoxy forms were consistent with coordination of the heme iron to histidine on the proximal side and binding of the CO without electrostatic interaction with the heme distal pocket. EcDos was unusual in having predominantly hexacoordinate heme iron in the deoxy and met forms. Despite an evident lack of CO interaction with the EcDos heme pocket, relatively low Fe O(2) (562 cm(-1)) and N-O (1576 cm(-1)) stretching frequencies indicated that strong polar interactions with that heme distal pocket are possible for highly bent ligands such as O(2) or NO. None of the newly studied NO adducts exhibited evidence of the Fe-His rupture and pentacoordination previously noted for Sinorhizobium meliloti FixL. A low Fe-His stretching frequency, formerly interpreted as a strained Fe-His bond, and the slow association of O(2) with S. meliloti FixL failed to correlate with the newly studied proteins having low association rate or low equilibrium association constants for binding of O(2). We conclude that although heme-PASs share some features, they represent distinct signal transduction mechanisms. PMID- 11939777 TI - The 1.25 A resolution structure of the diheme NapB subunit of soluble nitrate reductase reveals a novel cytochrome c fold with a stacked heme arrangement. AB - The diheme cytochrome NapB constitutes the small subunit of a periplasmic nitrate reductase found in a wide variety of bacterial species, including pathogens. The NapB protein is essential in transferring electrons to the large catalytic subunit NapA, which subsequently reduces nitrate to nitrite. Here we present the crystal structure of a proteolyzed form of recombinant NapB from Haemophilus influenzae, which was determined by the multiple-wavelength anomalous dispersion (MAD) method at 1.25 A resolution. This structure shows an unprecedented fold, confirming that NapB proteins belong to a new class of cytochromes. The two heme groups have nearly parallel heme planes and are stacked at van der Waals distances with an iron-to-iron distance of only 9.9 A, two structural features that are also present in the split-Soret diheme cytochrome c from Desulfovibrio desulfuricans ATCC 27774, which is otherwise unrelated in the peptide chain folding pattern. The two propionate side chains on both heme groups are hydrogen bonded to each other, a structural characteristic that to date also has not been reported in any other heme protein. The propionates of one of the heme groups are pulled toward the interior of the molecule due to a salt bridge and a number of hydrogen bonds between the propionates and conserved residues. We propose a hypothetical but plausible model of the NapAB complex in which the four redox centers are positioned in a virtually linear configuration which spans a distance of nearly 40 A, suggesting an efficient pathway for the transfer of electrons from NapC, the physiological electron donor of NapB, to a nitrate molecule at the catalytic site of NapA. PMID- 11939778 TI - Structural dynamics of distal histidine replaced mutants of myoglobin accompanied with the photodissociation reaction of the ligand. AB - Protein dynamics observed by the transient grating (TG) method are studied for some site-directed mutants at the distal histidine of myoglobin (H64L, H64Q, H64V). The time profiles of the TG signals are very sensitive to the amino acid residue of the 64 position. It was found that the sensitivity is mostly caused by the different rates of the ligand escape from the protein to solvent and the magnitude of the molecular volume change. Several molecular origins of the volume difference between MbCO and Mb, such as the electrostatic interaction in the distal pocket, movement of helices, and distal water, are proposed. Interestingly, the volume difference between the CO-trapped Mb inside the protein interior and Mb is similar to that of the partial molar volume of CO in organic solvent. The effect of mutation on the nature of the CO trapped site is discussed. PMID- 11939780 TI - Altering the RNase H primer grip of human immunodeficiency virus reverse transcriptase modifies cleavage specificity. AB - Recent crystallographic data suggest that conserved residues in the connection subdomain and C-terminal ribonuclease H (RNase H) domain of human immunodeficiency virus type 1 reverse transcriptase (HIV-1 RT) contact the nascent DNA primer and modulate the trajectory of the template relative to the RNase H catalytic center. Within the RNase H domain, these residues include Thr473, Glu475, Lys476, Tyr501, and Ile505, while His539 and Asn474 interact with the scissile phosphate of the RNA template. Amino acid substitutions at several of these positions were evaluated in the context of hydrolysis of nonspecific RNA DNA hybrids and substrates mimicking specific RNase H-mediated events. With the exception of mutant I505G, which exhibited a dimerization defect, substituting alanine at positions 473-476 and 501 had minimal consequences for DNA synthesis on duplex and hybrid DNA and RNA substrates. In contrast, the efficiency with which most mutants catalyzed polymerization-independent RNase H cleavage was sharply reduced. This deficiency was more pronounced when mutant enzymes were challenged to process the (+) strand polypurine tract (PPT) primer from either (+) RNA or a PPT/(+) DNA RNA/DNA chimera. Reduced polymerization-independent RNase H activity also significantly influenced the rate of DNA strand transfer, suggesting the donor template must be reduced in size below 13 nt before this event proceeds. PMID- 11939779 TI - Exploring the structure and activity of haloalkane dehalogenase from Sphingomonas paucimobilis UT26: evidence for product- and water-mediated inhibition. AB - The hydrolysis of haloalkanes to their corresponding alcohols and inorganic halides is catalyzed by alpha/beta-hydrolases called haloalkane dehalogenases. The study of haloalkane dehalogenases is vital for the development of these enzymes if they are to be utilized for bioremediation of organohalide contaminated industrial waste. We report the kinetic and structural analysis of the haloalkane dehalogenase from Sphingomonas paucimobilis UT26 (LinB) in complex with each of 1,2-dichloroethane and 1,2-dichloropropane and the reaction product of 1-chlorobutane turnover. Activity studies showed very weak but detectable activity of LinB with 1,2-dichloroethane [0.012 nmol s(-1) (mg of enzyme)(-1)] and 1,2-dichloropropane [0.027 nmol s(-1) (mg of enzyme)(-1)]. These activities are much weaker compared, for example, to the activity of LinB with 1 chlorobutane [68.2 nmol s(-1) (mg of enzyme)(-1)]. Inhibition analysis reveals that both 1,2-dichloroethane and 1,2-dichloropropane act as simple competitive inhibitors of the substrate 1-chlorobutane and that 1,2-dichloroethane binds to LinB with lower affinity than 1,2-dichloropropane. Docking calculations on the enzyme in the absence of active site water molecules and halide ions confirm that these compounds could bind productively. However, when these moieties were included in the calculations, they bound in a manner similar to that observed in the crystal structure. These data provide an explanation for the low activity of LinB with small, chlorinated alkanes and show the importance of active site water molecules and reaction products in molecular docking. PMID- 11939781 TI - Contribution of buried lysine residues to the oligomerization specificity and stability of the fos coiled coil. AB - Coiled coils comprise two or more helices characterized by a heptad repeat of amino acids denoted a through g. The buried a and d positions are usually occupied by hydrophobic residues. Fos dimerizes via a coiled coil (leucine zipper) with Jun family members to form the transcription factor AP-1. Fos homodimers are relatively unstable due to unfavorable interhelical electrostatic interactions within the Fos two-stranded coiled coil. The Fos coiled coil contains two polar position a Lys residues (Lys 16 and Lys 30 of Fos-p1, a peptide corresponding to the coiled-coil domain of v-Fos). Lys 16 and Lys 30 of Fos-p1 were replaced individually and together with the unnatural amino acid norleucine (2-aminohexanoic acid), which corresponds to a deletion of the Lys epsilon-amino group. The midpoint of thermal denaturation (T(m)) of Fos-p1 (10 microM) is 30 degrees C at pH 7. The Lys 16 --> Nle variant forms predominantly homodimers that are relatively unstable (T(m) = 46 degrees C). The Lys 30 --> Nle variant forms a stable homotetramer (T(m) = 60 degrees C). The Lys 16/Lys 30 --> Nle variant forms a very stable homotetramer (T(m) = 80 degrees C). The results show that (i) the effects of buried position a Lys residues on coiled-coil oligomerization are context dependent and (ii) electrostatic destabilization of the Fos homodimer can be mitigated by an oligomerization switch moderated by a single buried Lys residue. PMID- 11939782 TI - Molecular architecture of the thylakoid membrane: lipid diffusion space for plastoquinone. AB - We have determined the stoichiometric composition of membrane components (lipids and proteins) in spinach thylakoids and have derived the molecular area occupied by these components. From this analysis, the lipid phase diffusion space, the fraction of lipids located in the first protein solvation shell (boundary lipids), and the plastoquinone (PQ) concentration are derived. On the basis of these stoichiometric data, we have analyzed the motion of PQ between photosystem (PS) II and cytochrome (cyt.) bf complexes in this highly protein obstructed membrane (protein area about 70%) using percolation theory. This analysis reveals an inefficient diffusion process. We propose that distinct structural features of the thylakoid membrane (grana formation, microdomains) could help to minimize these inefficiencies and ensure a non-rate limiting PQ diffusion process. A large amount of published evidence supports the idea that higher protein associations exist, especially in grana thylakoids. From the quantification of the boundary lipid fraction (about 60%), we conclude that protein complexes involved in these associations should be spaced by lipids. Lipid-spaced protein aggregations in thylakoids are qualitatively different to previously characterized associations (multisubunit complexes, supercomplexes). We derive a hierarchy of protein and lipid interactions in the thylakoid membrane. PMID- 11939783 TI - Photoaffinity labeling of human retinoid X receptor beta (RXRbeta) with 9-cis retinoic acid: identification of phytanic acid, docosahexaenoic acid, and lithocholic acid as ligands for RXRbeta. AB - We utilized [20-methyl-(3)H]-9-cis-retinoic acid ([(3)H]9-cis-RA) as a direct photoaffinity probe for the characterization of human recombinant retinoid X receptor beta protein (RXRbeta). The photoaffinity labeling was light- and concentration-dependent, saturable, and protected by unlabeled 9-cis-RA in a concentration-dependent manner, indicating that binding occurred in the RXR retinoid binding site. all-trans-Retinoic acid (atRA) did not affect labeling with the 9-cis derivative, confirming that atRA does not compete for the 9-cis-RA binding site. Several retinoid, fatty acid, and bile acid ligands were evaluated for their ability to recognize the 9-cis-RA binding site. Retinol, atRA glucuronide, 13-cis-RA, dolichol, 5,6-epoxy-RA, and vitamin D(3) did not compete for the 9-cis-RA binding site. However, the saturated diterpenoid phytanic acid (PA) and docosahexaenoic acid, which have been recently shown to activate the nuclear receptor, RXR, competed with 9-cis-RA labeling, showing high affinity for the 9-cis-RA binding site. Oleic acid, arachidonic acid, and butyric acid did not interact. However, the bile acid lithocholic acid competed efficiently with 9-cis RA for the binding site. These data validated the photoaffinity assay as an excellent system for the identification and evaluation of ligands for RXR. PMID- 11939784 TI - The p58 subunit of human DNA primase is important for primer initiation, elongation, and counting. AB - The p58 subunit of human DNA primase contains a region, M288-K344, that is homologous to part of the 8 kDa domain of DNA polymerase beta. Since regions of a protein that are highly conserved evolutionarily often play important catalytic functions, we examined the effects of mutating this region of the p58 subunit on primase activity. Deleting M288-L313 of the p58 subunit results in a protein that binds to the primase p49 subunit but cannot support primer synthesis on any template when assays only contain Mg(2+) as the divalent metal. Including Mn(2+), a metal that stimulates initiation of primer synthesis, in the assays now allows the enzyme to synthesize primers at a rate only moderately lower than that of the wild-type enzyme on templates consisting solely of deoxycytidylates. While the enzyme is active under these conditions, it has lost the ability to synthesize primers of defined length (i.e., count). Alanine scanning mutagenesis of charged residues in this region revealed three amino acids, R302, R306, and K314, that play important roles in both primer initiation and translocation. Conversion of these residues to alanine interfered with initiation and significantly decreased the processivity of primase. Together, these studies indicate that this "pol beta like" region of p58 is important for three distinct aspects of primer synthesis:; initiation, translocation, and counting. The implications of these results with respect to the biological role of the p58 subunit and the mechanism of primer synthesis are discussed. PMID- 11939785 TI - Genetic remodeling and transcriptional remodeling of subtelomeric heterochromatin are different. AB - The structure, the extension, and the regulatory functions of telomeric and subtelomeric heterochromatin are not completely understood partly due to the difficulty of separating structural from functional features. We have previously observed that genetic alterations of telomeric heterochromatin components relieve transcriptional silencing. We have developed an analytical system allowing the separate determination of the effects of transcription and of genetic alterations on the subtelomeric structures. The uncoupled analysis, performed on the left extremity of chromosome III of Saccharomyces cerevisiae, consists of genetic dissections, induction of transcription of a resident gene, and chromatin analysis. The results allow (i) the determination of the precise localization and of the extension of heterochromatin (here from 0.9 to 2.6 kb from the innermost extremity of the C(1-3)A tract) and (ii) the definition of the transcription and of the genetically induced chromatin remodelings and of their marked differences, thus allowing (iii) specific analyses of the structural effects of the genetic modification of the heterochromatin components. PMID- 11939786 TI - Role of group IIa and group V secretory phospholipases A(2) in the metabolism of lipoproteins. Substrate specificities of the enzymes and the regulation of their activities by sphingomyelin. AB - Although many isoforms of secretory phospholipases A(2) (sPLA(2)) are known to be secreted by various inflammatory cells, and are present in plasma, their role in lipoprotein metabolism is unknown. We studied the in vitro hydrolysis of lipoprotein phospholipids by group IIa and group V sPLA(2), two structurally related enzymes with differing phospholipid specificities. The group V sPLA(2) was about 30 times more efficient than the group IIa enzyme in the hydrolysis of lipoprotein phosphatidylcholine (PC), and both enzymes were more active on high density liporotein (HDL) than on low density lipoprotein (LDL). The lower activity on LDL appears to be due to the higher sphingomyelin (SPH) concentration in this lipoprotein. PC hydrolysis in lipoproteins was stimulated significantly by enzymatic depletion of their SPH. The hydrolysis of PC in liposomes was inhibited by the incorporation of SPH, and this inhibition was reversed by treatment with sphingomyelinase. The incorporation of ceramide, on the other hand, stimulated the sPLA(2) activity significantly. Unlike most sPLA(2), which show no fatty acid preference, group V sPLA(2) released disproportionately more linoleate, and less arachidonate from lipoproteins. These studies show that group V sPLA(2) is physiologically more important than group IIa enzyme in lipoprotein metabolism, that the sPLA(2) activities are regulated by sphingomyelin and ceramide, and that the pathological effects of sPLA(2) may not be mediated through stimulation of eicosanoid synthesis. PMID- 11939787 TI - Receptor-associated protein facilitates proper folding and maturation of the low density lipoprotein receptor and its class 2 mutants. AB - Familial hypercholesterolemia is the consequence of various mutations in the low density lipoprotein receptor (LDLR). In the current study, we show that a specialized molecular chaperone, the receptor-associated protein (RAP), promotes proper folding and subsequent exocytic trafficking of the wild-type LDLR and several of its class 2 mutants. Co-immunoprecipitation with anti-RAP antibody demonstrates that RAP interacts with the LDLR. Kinetic analyses of LDLR posttranslational folding and maturation in the absence or presence of RAP coexpression show that RAP prevents aggregation and promotes the maturation of the LDLR. Additionally, depletion of Ca(2+) in intact cells impairs LDLR folding, and coexpression of RAP partially corrects this misfolding. Finally, we show that the increased mature cell surface LDLR in the presence of RAP coexpression is functional in its ability to endocytose and degrade (125)I-LDL. Taken together, our results show that the folding, trafficking, and maturation of the LDLR and its class 2 mutants are promoted by RAP. PMID- 11939788 TI - Sterol efflux to apolipoprotein A-I originates from caveolin-rich microdomains and potentiates PDGF-dependent protein kinase activity. AB - The kinetics of sterol efflux from human aortic smooth muscle cells equilibrated with a [(3)H]benzophenone-modified photoactivable free cholesterol analogue ((3)H FCBP) did not differ significantly from those labeled with free cholesterol ((3)H FC). Trypsin digestion of caveolin cross-linked by photoactivation of FCBP led to association of radiolabel in a single low molecular weight fraction, indicating relative structural homogeneity of caveolin-bound sterol. These findings were used to investigate the organization of sterols in caveolae and the ability of these domains to transfer sterols to apolipoprotein A-I (apo A-I), the major protein of human plasma high-density lipoproteins (HDL). During long-term (4-5 h) incubation with apo A-I, caveolin-associated (3)H-FC and (3)H-FCBP decreased, in parallel with an increase in apo A-I-associated sterol. Assay of caveolin associated labeled sterols indicated that caveolae were a major source of sterol lost from the cells during HDL formation. Short-term changes of sterol distribution in caveolae were assayed using platelet-derived growth factor (PDGF). PDGF was without effect on FC efflux in the absence of apo A-I, but when apo A-I was present, PDGF increased FC efflux approximately 3-fold beyond the efflux rate catalyzed by apo A-I alone. At the same time, caveolin-associated FC decreased, and PDGF-dependent protein kinase activity was stimulated. Parallel results were obtained with (3)H-FCBP-equilibrated cells, in which apo A-I potentiated a PDGF-mediated reduction of radiolabel cross-linked to caveolin following photoactivation. These results suggest that sterols within caveolae are mobile and selectively transferred to apo A-I. They also suggest a novel role for sterol efflux in amplifying PDGF-mediated signal transduction. PMID- 11939789 TI - Inactivation of the PKR protein kinase and stimulation of mRNA translation by the cellular co-chaperone P58(IPK) does not require J domain function. AB - P58(IPK) was discovered as an inhibitor of the interferon-induced, protein kinase, PKR. Upon virus infection, PKR can, as part of the host defense system, inhibit mRNA translation by phosphorylating the alpha subunit of protein synthesis eukaryotic initiation factor 2 (eIF-2alpha). We previously found that influenza virus recruits the cellular P58(IPK) co-chaperone to inhibit PKR activity and thus facilitate viral protein synthesis. P58(IPK) contains nine tetratricopeptide repeat (TPR) motifs in addition to the highly conserved J domain found in all DnaJ chaperone family members. To define the role of molecular chaperones in regulating cell growth in addition to PKR regulation, we performed a detailed analysis of the P58(IPK) J domain. Using growth rescue assays, we found that the P58(IPK) J domain substituted for the J domains of other DnaJ proteins, including DnaJ in Escherichia coli and Ydj1 in Saccharomyces cerevisiae. This is the first time a cellular J domain from a mammalian DnaJ family member was shown to be functional in both prokaryotic DnaJ and eukaryotic Ydj1 constructs. Furthermore, point mutations within the conserved HPD residue cluster of the P58(IPK) J domain disrupted P58(IPK) J function including stimulation of ATPase activity of Hsp70. However, the P58(IPK) HPD mutants still inhibited PKR activity and thus supported cell growth in a yeast rescue assay. Overexpression of the HPD mutants of P58(IPK), similar to their wild-type counterpart, also stimulated mRNA translation in a mammalian cell system. Taken together, our data necessitate a model of P58(IPK) inhibition of PKR kinase activity and stimulation of mRNA translation, which does not require classical J domain function found in the DnaJ molecular chaperone family. PMID- 11939790 TI - Importance of hydrophobic matching for spontaneous insertion of a single-spanning membrane protein. AB - In this study, we have investigated the effect of hydrophobic mismatch between the thickness of the membrane and a transmembrane segment of a protein that directly inserts into the membrane bilayer. For this purpose we used mutants of the single-spanning Pf3 coat protein that can spontaneously insert into Escherichia coli membrane vesicles and large unilamellar vesicles (LUVs). The thickness of the liposomal bilayer could be altered by using lipids with different acyl chain lengths or by incorporation of cholesterol. The insertion efficiency of the protein clearly depended on the bilayer thickness, with most efficient insertion under hydrophobic matching conditions. To discriminate between effects of length and hydrophobicity, mutants with different synthetic transmembrane segments were constructed. These mutants inserted into LUVs in a mismatch-dependent manner. However, in particular for longer and less hydrophobic mutants, most efficient insertion was generally observed in thinner bilayers than expected on the basis of hydrophobic matching. PMID- 11939791 TI - The p11 subunit of annexin II heterotetramer is regulated by basic carboxypeptidase. AB - The Ca(2+)-dependent phospholipid-binding protein annexin II heterotetramer (AIIt) is composed of two copies of annexin II and a p11 dimer. The interaction of the carboxyl-terminal lysine residues of the p11 subunit of AIIt with the lysine-binding kringle domains of plasminogen is believed to play a key role in plasminogen binding and stimulation of the tPA-catalyzed cleavage of plasminogen to plasmin. In the current report, we show that AIIt-stimulated plasminogen activation is regulated by basic carboxypeptidases, in vitro. The incubation of AIIt with a 1/400 molar ratio of carboxypeptidase B for periods as short as 2 min resulted in a significant loss in AIIt-stimulated plasminogen activation. Carboxypeptidase B (CpB) as well as thrombin-activated fibrinolysis inhibitor (TAFIa) and carboxypeptidase N (CpN) rapidly reduced AIIt-stimulated plasminogen activation by 80%. The molar ratio of carboxypeptidase/AIIt for half-maximal inhibition of AIIt was 1/4700, 1/700, and 1/500 for CpB, TAFIa, and CpN, respectively. Treatment of AIIt with carboxypeptidase resulted in loss of both carboxyl-terminal lysine residues from the p11 subunit, which correlated with a decrease in the k(cat) and an increase in the K(m) for plasminogen activation. The data reveal a novel mechanism for the regulation of AIIt-stimulated plasminogen activation. PMID- 11939792 TI - Mucopolysaccharidosis type VI (Maroteaux-Lamy syndrome): a Y210C mutation causes either altered protein handling or altered protein function of N acetylgalactosamine 4-sulfatase at multiple points in the vacuolar network. AB - The lysosomal hydrolase N-acetylgalactosamine 4-sulfatase (4-sulfatase) is required for the degradation of the glycosaminoglycan substrates dermatan and chondroitin sulfate. A 4-sulfatase deficiency results in the accumulation of undegraded substrate and causes the severe lysosomal storage disorder mucopolysaccharidosis type VI (MPS VI) or Maroteaux-Lamy syndrome. A wide variation in clinical severity is observed between MPS VI patients and reflects the number of different 4-sulfatase mutations that can cause the disorder. The most common 4-sulfatase mutation, Y210C, was detected in approximately 10% of MPS VI patients and has been associated with an attenuated clinical phenotype when compared to the archetypical form of MPS VI. To define the molecular defect caused by this mutation, Y210C 4-sulfatase was expressed in Chinese hamster ovary (CHO-K1) cells for protein and cell biological analysis. Biosynthetic studies revealed that Y210C 4-sulfatase was synthesized at a comparable molecular size and amount to wild-type 4-sulfatase, but there was evidence of delayed processing, traffic, and stability of the mutant protein. Thirty-three percent of the intracellular Y210C 4-sulfatase remained as a precursor form, for at least 8 h post labeling and was not processed to the mature lysosomal form. However, unlike other 4-sulfatase mutations causing MPS VI, a significant amount of Y210C 4-sulfatase escaped the endoplasmic reticulum and was either secreted from the expression cells or underwent delayed intracellular traffic. Sixty-seven percent of the intracellular Y210C 4-sulfatase was processed to the mature form (43, 8, and 7 kDa molecular mass forms) by a proteolytic processing step known to occur in endosomes-lysosomes. Treatment of Y210C CHO-K1 cells with the protein stabilizer glycerol resulted in increased amounts of Y210C 4-sulfatase in endosomes, which was eventually trafficked to the lysosome after a long, 24 h chase time. This demonstrated delayed traffic of Y210C 4-sulfatase to the lysosomal compartment. The endosomal Y210C 4-sulfatase had a low specific activity, suggesting that the mutant protein also had problems with stability. Treatment of Y210C CHO-K1 cells with the protease inhibitor ALLM resulted in an increased amount of mature Y210C 4-sulfatase localized in lysosomes, but this protein had a very low level of activity. This indicated that the mutant protein was being inactivated and degraded at an enhanced rate in the lysosomal compartment. Biochemical analysis of Y210C 4-sulfatase revealed a normal pH optimum for the mutant protein but demonstrated a reduced enzyme activity with time, also consistent with a protein stability problem. This study indicated that multiple subcellular and biochemical processes can contribute to the biogenesis of mutant protein and may in turn influence the clinical phenotype of a patient. In MPS VI patients with a Y210C allele, the composite effect of different stages of intracellular processing/handling and environment has been shown to cause a reduced level of Y210C 4-sulfatase protein and activity, resulting in an attenuated clinical phenotype. PMID- 11939793 TI - Structure-activity relationship of hydroxamate-based inhibitors on the secretases that cleave the amyloid precursor protein, angiotensin converting enzyme, CD23, and pro-tumor necrosis factor-alpha. AB - Multiple proteins are proteolytically shed from the membrane, including the amyloid precursor protein (APP) involved in Alzheimer's disease, the blood pressure regulating angiotensin converting enzyme (ACE), the low affinity IgE receptor CD23, and the inflammatory cytokine tumor necrosis factor-alpha (TNF alpha). The inhibitory effect of a range of hydroxamic acid-based compounds on the secretases involved in cleaving and releasing these four proteins has been examined to build up a structure-activity relationship. Compounds have been identified that can discriminate between TNF-alpha convertase and the other three secretases (compound 15), between the shedding of CD23 and the shedding of APP and ACE (compound 21), and between the secretases and matrix metalloproteinase-1 (compound 22). The structure-activity relationship for the APP alpha-secretase and the ACE secretase were remarkably similar, and both secretases were activated in whole cell systems by the serine proteinase inhibitor 3,4-dichloroisocoumarin. The basal and carbachol-stimulated shedding of APP and ACE from human SH-SY5Y neuroblastoma cells could not be differentiated by any of the hydroxamate compounds, implying that the same or very similar activities are involved in the constitutive and regulated shedding of these two proteins. By utilizing a key discriminatory compound (compound 15) that potently inhibits TNF-alpha convertase but not alpha-secretase, we show that TNF-alpha convertase is not involved in the regulated shedding of APP from human neuronal cells. The compounds reported here will be useful in future studies aimed at identifying and validating candidate secretases. PMID- 11939794 TI - Lysophosphatidic acid and receptor-mediated activation of endothelial nitric oxide synthase. AB - Both lysophosphatidic acid (LPA) and sphingosine-1-phosphate (S1P) are platelet derived phospholipids that elicit diverse biological responses. In endothelial cells, S1P stimulates the EDG-1 receptor-mediated activation of the endothelial isoform of nitric oxide synthase (eNOS), but the role of LPA in eNOS regulation is less well understood. We now report that LPA treatment of bovine aortic endothelial cells (BAEC) activates eNOS enzyme activity in a pathway that involves phosphorylation of eNOS on serine 1179 by protein kinase Akt. In contrast to the cellular responses elicited by S1P in COS-7 cells, LPA can stimulate the activation of eNOS and Akt independently of EDG-1 receptor transfection. LPA-stimulated enzyme activation was significantly attenuated in an eNOS mutant lacking the site that is phosphorylated by kinase Akt (eNOS S1179A). In BAEC, activation of eNOS by LPA is completely blocked by pertussis toxin, by the intracellular calcium chelator BAPTA (1,2-bis(aminophenoxy) ethane-N,N,N',N' tetraacetic acid), and by the phosphoinositide 3-kinase (PI3-K) inhibitor wortmannin, but is unaffected by U0126, an inhibitor of mitogen-activated protein (MAP) kinase pathways. Analysis of the LPA dose response for eNOS activation reveals an EC(50) of approximately 40 nM, a concentration well below the potency of LPA at the EDG-1 receptor. Taken together, these results indicate that LPA potently activates eNOS in BAEC in a pathway distinct from the EDG-1 receptor, but mediated by a similar receptor-mediated pathway dependent on pertussis toxin sensitive G proteins and involving activation of the PI3-K/Akt pathway. These studies have identified a role for the phospholipid LPA in eNOS activation, and point out the complementary role of distinct platelet-derived lipids in endothelial signaling pathways. PMID- 11939795 TI - Contribution of regions distal to glycine-160 to the anticoagulant activity of tissue factor pathway inhibitor. AB - The functions of the first two Kunitz domains of tissue factor pathway inhibitor (TFPI) are well defined as active site-directed inhibitors of factor VIIa and factor Xa. The anticoagulant properties of the third Kunitz domain and C-terminal region were probed using altered forms of TFPI. TFPI-160 contains the first two Kunitz domains. K1K2C contains the first two Kunitz domains and the basic C terminus. Neither TFPI-160 nor K1K2C contains the third Kunitz domain. In amidolytic assays containing calcium, TFPI-160 is a less potent inhibitor of factor Xa than TFPI. However, addition of the C-terminus in K1K2C nearly restores inhibitory activity to that of TFPI, indicating that the third Kunitz domain is not required for direct inhibition of factor Xa. When compared in assays containing phospholipids and factor Va, K1K2C and TFPI-160 are poor inhibitors compared to TFPI, demonstrating that the third Kunitz domain is required for the full anticoagulant activity of TFPI. TFPI was further characterized in amidolytic assays performed with Gla-domainless factor Xa and in prothrombin activation assays using submicellar concentrations of short-chain phospholipids (C6PS). TFPI and K1K2C are worse inhibitors of Gla-domainless factor Xa, compared to wild-type factor Xa, while TFPI-160 inhibits both forms of factor Xa equally, suggesting a C-terminus/Gla domain interaction. TFPI is a potent inhibitor of thrombin generation by prothrombinase assembled with C6PS, while TFPI-160 and K1K2C are not. Conversely, TFPI does not inhibit prothrombin activation by prothrombinase assembled on a two-dimensional lipid bilayer. Together, the data indicate that the region between Gly-160 and the end of the third Kunitz domain contributes to TFPI function by orienting the second Kunitz domain so that it can bind the active site of phospholipid-associated factor Xa prior to prothrombinase assembly and/or by slowing formation of the prothrombinase complex. PMID- 11939796 TI - Evidence that serpin architecture intrinsically supports papain-like cysteine protease inhibition: engineering alpha(1)-antitrypsin to inhibit cathepsin proteases. AB - The closely related serpins squamous cell carcinoma antigen-1 and -2 (SCCA-1 and 2, respectively) are capable of inhibiting cysteine proteases of the papain superfamily. To ascertain whether the ability to inhibit cysteine proteases is an intrinsic property of serpins in general, the reactive center loop (RCL) of the archetypal serine protease inhibitor alpha(1)-antitrypsin was replaced with that of SCCA-1. It was found that this simple substitution could convert alpha(1) antitrypsin into a cysteine protease inhibitor, albeit an inefficient one. The RCL of SCCA-1 is three residues longer than that of alpha(1)-antitrypsin, and therefore, the effect of loop length on the cysteine protease inhibitory activity was investigated. Mutants in which the RCL was shortened by one, two, or three residues were effective inhibitors with second-order rate constants of 10(5) 10(7) M(-)(1) s(-)(1). In addition to loop length, the identity of the cysteine protease was of considerable importance, since the chimeric molecules inhibited cathepsins L, V, and K efficiently, but not papain or cathepsin B. By testing complexes between an RCL-mimicking peptide and the mutants, it was found that the formation of a stable serpin-cysteine protease complex and the inhibition of a cysteine protease were both critically dependent on RCL insertion. The results strongly indicate that the serpin body is intrinsically capable of supporting cysteine protease inhibition, and that the complex with a papain-like cysteine protease would be expected to be analogous to that seen with serine proteases. PMID- 11939797 TI - Kinetic and microcalorimetric analysis of substrate and cofactor interactions in epoxyalkane:CoM transferase, a zinc-dependent epoxidase. AB - Epoxyalkane:CoM transferase (EaCoMT) is a key enzyme of bacterial propylene metabolism, catalyzing the nucleophilic attack of coenzyme M (CoM, 2 mercaptoethanesulfonic acid) on epoxypropane to form the thioether conjugate 2 hydroxypropyl-CoM. The biochemical and molecular properties of EaCoMT suggest that the enzyme belongs to the family of alkyltransferase enzymes for which Zn plays a key role in activating an organic thiol substrate for nucleophilic attack on an alkyl-donating substrate. In the present work, the role of Zn in the EaCoMT catalyzed reactions is established by removing Zn from EaCoMT, resulting in loss of catalytic activity that was restored upon addition of Zn back to the enzyme, and by expressing an inactive and Zn-deficient form of the enzyme that was activated by addition of ZnCl(2) or CoCl(2). Site-directed mutagenesis of one of the predicted Zn ligands (C220A) resulted in the formation of a largely catalytically inactive protein (0.06% of wild-type activity) that, when purified, contained a substoichiometric complement of Zn. EaCoMT was kinetically characterized and found to follow a random sequential mechanism with kinetic parameters K(m,epoxypropane) = 1.8 microM, K(m,CoM) = 34 microM, and k(cat) = 6.5 s(-1). The CoM analogues 2-mercaptopropionate, 2-mercaptoethanol, and cysteine substituted poorly for CoM as the thiol substrate, with specific rates of epoxyalkane conjugation that were at best 0.6% of the CoM-dependent rate, while ethanethiol, propanethiol, glutathione, homocysteine, and lipoic acid provided no activity. 2-Mercaptoethanol was a weak competitive inhibitor vs CoM with a K(I) of 192 mM. Isothermal titration calorimetry was used to investigate the thermodynamic binding determinants for the interaction of CoM and analogues with holo, Zn-deficient, and C220A EaCoMT variants. The stoichiometry of CoM binding correlated directly with the Zn content rather than monomer content of protein samples, reinforcing the importance of Zn in CoM binding. The binding of CoM to EaCoMT occurred with DeltaG = -7.5 kcal/mol (K(d) = 3.8 microM) and was driven by a large release of enthalpy. The thermodynamic contributors (K(a), DeltaG, DeltaH, DeltaS) to the individual binding of CoM, ethanesulfonate, and ethanethiol were determined and used to assess the contributions of the thiol, alkyl, and sulfonate moieties to total binding energy in the E x CoM binary complex. PMID- 11939798 TI - The temperature dependence of P680(+) reduction in oxygen-evolving photosystem II. AB - The temperature dependence for the reduction of the oxidized primary electron donor P680(+) by the redox active tyrosine Y(Z) has been studied in oxygen evolving photosystem II preparations from spinach. The observed temperature dependence is found to vary markedly with the S-state of the manganese cluster. In the higher oxidation states, S(2) and S(3), sub-microsecond P680(+) reduction exhibits activation energies of about 260 meV. In contrast, there is only a small temperature dependence for the sub-microsecond reaction in the S(0) and S(1) states (an activation energy of approximately 50 meV). Slower microsecond components of P680(+) reduction show an activation energy of about 250 meV which, within experimental error, is independent of the oxidation state of the Mn cluster. By combining these values with measurements of DeltaG for electron transfer, the reorganization energies for each component of P680(+) reduction have been calculated. High activation and reorganization energies are found for sub-microsecond P680(+) reduction in S(2) and S(3), demonstrating that these electron transfers are coupled to significant reorganization events which do not occur in the presence of the lower S-states. One interpretation of these results is that there is an increase in the net charge on the manganese cluster on the S(1) to S(2) transition which acts as a barrier to electron transfer in the higher S-states. This argues against the electroneutrality requirement for some models of the function of the manganese cluster and hence against a role for Y(Z) as a hydrogen abstractor on all S-state transitions. An alternative or additional possibility is that there are proton (or other ion) motions in the sub microsecond phases in S(2) and S(3) which contribute to the large reorganization energies observed, these motions being absent in the S(0) and S(1) states. Indeed charge accumulation may directly cause the increased reorganization energy. PMID- 11939799 TI - Characterization of an iron-sulfur cluster assembly protein (ISU1) from Schizosaccharomyces pombe. AB - Genetic studies of bacteria and eukaryotes have led to identification of several gene products that are involved in the biosynthesis of protein-bound iron-sulfur clusters. One of these proteins, ISU, is homologous to the N-terminus of bacterial NifU. The mature forms of His-tagged wild-type and D37A Schizosaccharomyces pombe ISU1 were cloned and overexpressed as inclusion bodies in Escherichia coli. The recombinant D37A protein was purified under denaturing conditions and subsequently reconstituted in vitro. By use of a 5-fold excess of iron and sulfide the reconstituted product was found to be red-brown in color, forming a homodimer of 17 kDa per subunit with approximately two iron atoms per monomer determined by protein and iron quantitation. UV-vis absorption and Mossbauer spectroscopies (delta = 0.29 +/- 0.05 mm/s; DeltaE(Q) = 0.59 +/- 0.05 mm/s) were used to characterize D37A ISU1 and show the presence of [2Fe-2S](2+) clusters in each subunit. Formation of the holo form of wild-type ISU1 was significantly less efficient using the same reconstitution conditions and is consistent with prior observations that the D37A substitution can stabilize protein-bound clusters. Relative to the human homologue, the yeast ISU is significantly less soluble at ambient temperatures. In both cases the native ISU1 is more sensitive to proton-mediated degradation relative to the D37A derivative. The lability of this family of proteins relative to [2Fe-2S] bearing ferredoxins most likely is of functional relevance for cluster transfer chemistry. Mossbauer parameters obtained for wild-type ISU1 (delta = 0.31 +/- 0.05 mm/s; DeltaE(Q) = 0.64 +/- 0.05 mm/s) were similar to those obtained for the D37A derivative. Cluster transfer from ISU1 to apo Fd is demonstrated: the first example of transfer from an ISU-type protein. A lower limit for k(2) of 80 M(-1) min(-1) was established for WT cluster transfer and a value of 18 M(-1) min(-1) for the D37A derivative. Finally, we have demonstrated through cross-linking studies that ferredoxin, an electron-transport protein, forms a complex with ISU1 in both apo and holo states. Cross-linking of holo ISU1 with holo Fd is consistent with a role for redox chemistry in cluster assembly and may mimic the intramolecular complex already defined in NifU. PMID- 11939800 TI - Effects of DNA structure on oxopropenylation by the endogenous mutagens malondialdehyde and base propenal. AB - Malondialdehyde (MDA) and nucleobase propenals can transfer oxopropenyl groups to guanine residues of DNA to yield pyrimodopurinone (M(1)G) adducts. The DNA structural requirements for reaction with alpha,beta-unsaturated aldehydes were explored. We found that single-stranded DNA is more sensitive to oxopropenylation than double-stranded DNA, and supercoiled plasmid DNA is more sensitive than linearized plasmid DNA. Increasing ionic strength inhibits oxopropenylation, especially by adenine propenal. The intercalating agents ethidium bromide and 9 aminoacridine enhanced oxopropenylation by severalfold. In contrast, actinomycin D, which both intercalates and binds in the minor groove, inhibited oxopropenylation. The anthracycline drugs daunorubicin and doxorubicin enhanced oxopropenylation by MDA up to 3-fold and by adenine propenal up to 7-fold in a concentration-dependent manner. The minor groove binders netropsin and distamycin inhibited oxopropenylation, but methyl green, a major groove binder, had little effect. These data suggest that steric access to the target nucleophile located in the minor groove of DNA is critical for adduct formation by the endogenous mutagens MDA and base propenals. PMID- 11939801 TI - Actinomycin D binds to d(TGTCATG) with 2:1 drug to duplex stoichiometry. AB - Despite the apparent single-stranded conformation and the absence of a GpC site, d(TGTCATTG) has been found to bind strongly to actinomycin D (ACTD) with 1:1 drug to strand binding stoichiometry. A hairpin binding model was speculated in which the planar phenoxazone chromophore inserts at the GTC site by pushing out the T base while the terminal G folds back to form a G small middle dotC base pair so that the 3'-sides of both G-bases stack on the opposite faces of the phenoxazone plane. However, it was also suggested that a slipped duplex binding with similar binding principle could also be operative at higher DNA concentrations. To support such a contention, ACTD binding studies were made with d(TGTCATG) and related oligomers. This heptamer differs from the parent octamer d(TGTCATTG) by a mere removal of a T-base which should result in an enhancement of dimeric duplex formation and a concomitant reduction in monomeric hairpin contribution. It was found that ACTD binds well to d(TGTCATG) with 1 drug to 1 strand (or 2 drugs to 1 duplex) binding stoichiometry. These results are consistent with a slipped duplex binding model in which a dimeric duplex is formed at the self-complementary CATG tetranucleotide sequence with extruding TGT ends. Two drug molecules are bound at both ends of the duplex by pushing out the T-bases of GTC's so that the opposite faces of each phenoxazone are stacked by the 3'-sides of the two G-bases on opposite strands. Such a model provides a ready explanation for the observed enhancement in ACTD binding to d(TGTCATGTC) and d(TGTCATGTCA), where additional base pairs at the ends will stabilize GTC/GTC binding sites, and to d(TGTCAATTG) in which two additional base pairs facilitate the slipped-duplex formation. The observed ACTD affinity reductions for oligomers containing GTTC instead of GTC are also consistent with the T-base displacement model. These findings greatly expand the repertoire of ACTD binding to DNA and may have important implications on understanding the transcription inhibitory activities of this drug. PMID- 11939802 TI - Requirement for GroEL/GroES-dependent protein folding under nonpermissive conditions of macromolecular crowding. AB - Macromolecular crowding is a critical parameter affecting the efficiency of cellular protein folding. Here we show that the proteins dihydrofolate reductase, enolase, and green fluorescent protein, which can fold spontaneously in diluted buffer, lose this ability in a crowded environment. Instead, they accumulate as soluble, protease-sensitive non-native species. Their folding becomes dependent on the complete GroEL/GroES chaperonin system and is not affected by trap-GroEL, indicating that folding has to occur in the chaperonin cavity with release of nativelike proteins into the bulk solution. In addition, we demonstrate that efficient folding in the chaperonin cavity requires ATP hydrolysis, as formation of ternary GroEL/GroES complexes with substrate proteins in the presence of ADP results only in very inefficient reactivation. However, protein refolding reactions using ADP-fluoroaluminate complexes, or single-ring GroEL and GroES under conditions where only a single round of ATP hydrolysis occurs, yield large amounts of refolded enzymes. Thus, the mode of initial ternary complex formation appears to be critical for subsequent productive release of substrate into the cavity under certain crowding conditions, and is only efficient when triggered by ATP hydrolysis. Our data indicate that stringent conditions of crowding can impart a stronger dependence of folding proteins on the assistance by chaperonins. PMID- 11939803 TI - Quantitative analysis of the relative contributions of donor acyl carrier proteins, acceptor ketosynthases, and linker regions to intermodular transfer of intermediates in hybrid polyketide synthases. AB - 6-Deoxyerythronolide B synthase (DEBS) is the modular polyketide synthase (PKS) responsible for the biosynthesis of 6-dEB, the aglycon core of the antibiotic erythromycin. The biosynthesis of 6-dEB proceeds in an assembly-line fashion through the six modules of DEBS, each of which catalyzes a dedicated set of reactions, such that the structure of the final product is determined by the arrangement of modules along the assembly line. This transparent relationship between protein sequence and enzyme function is common to all modular PKSs and makes these enzymes an attractive scaffold for protein engineering through module swapping. One of the fundamental issues relating to module swapping that still needs to be addressed is the mechanism by which intermediates are channeled from one module to the next. While it has been previously shown that short linker regions at the N- and C-termini of adjacent polypeptides play an important role in mediating intermodular transfer, the contributions of other protein-protein interactions have not yet been probed. Here, we investigate the roles of the linker interactions as well as the interactions between the donor acyl carrier protein (ACP) domain and the downstream ketosynthase (KS) domain in various contexts. Linker interactions and ACP-KS interactions make relatively equal contributions at the module 2-module 3 and the module 4-module 5 interfaces in DEBS. In contrast, modules 2 and 6 are more tolerant toward substrates presented by nonnatural ACP domains. This tolerance was exploited for engineering hybrid PKS-PKS and PKS-NRPS (nonribosomal peptide synthetase) junctions and suggests fundamental ground rules for engineering novel chimeric PKSs in the future. PMID- 11939804 TI - Axillary Fox-Fordyce disease treated with liposuction-assisted curettage. PMID- 11939805 TI - Diffuse dermal angiomatosis: a variant of reactive angioendotheliomatosis associated with peripheral vascular atherosclerosis. PMID- 11939807 TI - Patients' attitudes regarding physical characteristics of medical care providers in dermatologic practices. AB - OBJECTIVE: To assess the present attitudes of patients toward physicians' physical attributes. DESIGN: Written survey offered to all patients seen during a 1-week period. SETTING: Two outpatient dermatologic clinical practices (a county hospital and a private practice). PARTICIPANTS: Of 315 patients offered the survey, 275 agreed to complete it. MAIN OUTCOME MEASURES: Opinions regarding physicians' demographic characteristics and opinions regarding desirability of 19 and 18 appearance-related characteristics in male and female physicians, respectively. RESULTS: Analysis of the responses revealed 25 characteristics that were significantly desirable or undesirable (defined as being selected desirable or undesirable by at least 25% of respondents). Further analysis revealed that patients in a private practice setting typically had more polar opinions about providers' appearances than did patients from a large county hospital. Most patients had no preference with regard to the sex, age, or race of their medical care providers. Age and sex of the patient did not independently contribute significantly to patient preferences, as determined by cross-tabulation analysis. Clinic site (private practice vs county hospital clinic) alone was the sole or most important predictor of preferences in 13 of the 25 significant characteristics. CONCLUSIONS: Several characteristics of providers' dress and grooming were important to patients. There seemed to be little attitudinal change from similar studies performed 2 decades ago. Cognizance of these preferences may facilitate better interactions between medical care providers and patients. PMID- 11939808 TI - Interferon alfa-2a in the treatment of Behcet disease: a randomized placebo controlled and double-blind study. AB - OBJECTIVE: To determine the therapeutic efficacy of interferon alfa-2a in the treatment of Behcet disease. DESIGN: A randomized placebo-controlled and double blind study. SETTING: University referral center. PATIENTS: Fifty patients with Behcet disease were involved in the study. INTERVENTION: The patients were given interferon alfa-2a, 6 x 10(6) IU, subcutaneously 3 times per week or placebo for 3 months, and examined clinically at weekly intervals. MAIN OUTCOME MEASURES: For each mucocutaneous lesion and articular symptom, the mean frequency and duration were evaluated during the 3-month pretreatment, treatment, and follow-up periods. Pain for oral and genital ulcers was scored on a scale of 0 to 3. The ocular inflammatory score, the frequency of attacks, and changes in visual acuities for patients with ocular involvement were assessed before the study, at the end of treatment, and during the follow-up periods. In addition, overall responses at the end of the treatment period were graded as follows: complete remission, disappearance of all clinical signs and symptoms during treatment; partial remission, greater than a 50% decrease in the frequency, duration, and severity of pain for oral and genital ulcers and/or a decrease in the severity and frequency of ocular attacks; stable disease, less than a 50% change in the clinical signs and symptoms; and no effect or deterioration, ineffectiveness or worsening of clinical signs and symptoms. RESULTS: Twenty-three interferon alfa 2a- and 21 placebo-treated patients, ranging in age from 16 to 55 years (mean +/- SD age, 32.38 +/- 7.94 years), were evaluable for efficacy. Interferon alfa-2a treatment significantly decreased the duration (P=.02) and pain (P=.01) of oral ulcers and the frequency of genital ulcers (P=.03) and papulopustular lesions (P=.01). The mean frequency and duration of erythema nodosum-like lesions (P=.77 and.27, respectively), thrombophlebitis (P=.29 and.61, respectively), and articular symptoms (P=.92 and.74, respectively) also decreased. But there were no statistically significant differences. An improvement in the severity and the frequency of ocular attacks occurred in 5 of 6 patients in the interferon alfa-2a treated group and in 1 of 3 patients in the placebo-treated group. Of the 23 patients in the interferon alfa-2a-treated group, 15 responded to treatment (2 complete and 13 partial responses); and of the 21 patients in the placebo group, 3 responded to treatment (3 partial responses) (P<.005). CONCLUSION: Interferon alfa-2a is an effective alternative treatment for Behcet disease, particularly for the management of the mucocutaneous lesions of the disease. PMID- 11939809 TI - Release and activation of matrix metalloproteinase-9 during in vitro mechanical compression in hypertrophic scars. AB - OBJECTIVE: To investigate induction of matrix metalloproteinases (MMPs) during mechanical compression of hypertrophic scars. Mechanical pressure blocks hypertrophy inducted on extracellular matrix in scars by a mechanism that involves MMP-2 (gelatinase A) and MMP-9 (gelatinase B). DESIGN: We assayed conditioned media obtained from normotrophic and hypertrophic scars during 24 hours of in vitro mechanical compression using gelatin zymography. SETTING: Scars from various areas of the bodies of hospitalized patients. PATIENTS: We obtained 3 normotrophic and 7 hypertrophic biopsy specimens from 10 patients (5 men and 5 women). INTERVENTION: In vitro compression at a pressure of 35 mm Hg/cm(2) for 24 hours. MAIN OUTCOME MEASURES: Vitality of scars was analyzed by means of lactic dehydrogenase test; medium samples were collected for zymographic analysis of MMP activity. RESULTS: We found MMP-2 in basal (uncompressed) samples from normotrophic and hypertrophic scars. Mechanical compression induced MMP-9 release and activation (range, 86.7%-78.7%) in hypertrophic scars after 4 hours. CONCLUSION: Production, release, and activation of MMP-9 in hypertrophic scars could be an effector mechanism responsible for hypertrophy regression induced by mechanical compression. PMID- 11939810 TI - Successful treatment of acne vulgaris using a new method: results of a randomized vehicle-controlled trial of short-contact therapy with 0.1% tazarotene gel. AB - CONTEXT: Short-contact application of 0.1% tazarotene gel for acne was devised to minimize local adverse effects. Its efficacy and safety are unknown. OBJECTIVES: To assess acne improvement and tolerability during 12 weeks of short-contact treatment with 0.1% tazarotene gel vs a nonmedicated gel control. DESIGN: A randomized, masked, vehicle-controlled trial. SETTING: Outpatient facilities at an urban medical school and an affiliated suburban office practice. PARTICIPANTS: Ninety-nine volunteers with facial acne were enrolled; 81 completed the study. INTERVENTION: Thirty-three patients were randomly assigned to each of 3 groups: T + T applied 0.1% tazarotene gel twice daily, T + V applied 0.1% tazarotene gel once daily and vehicle gel once daily, and V + V applied vehicle gel twice daily. Patients adjusted the contact period as tolerated, between 30 seconds and 5 minutes per application. MAIN OUTCOME MEASURES: Acne efficacy by reduction in acne lesions, treatment success (50%-100% improvement in global response to treatment) and improvement in overall disease severity. Local adverse effects, scored from none to severe. RESULTS: By week 12, T + T and T + V achieved significantly greater improvement in acne than V + V based on mean percentage reduction in noninflammatory lesions (46% and 41% vs 2%; P =.002) and inflammatory lesions (38% and 34% vs 9%; P =.01), percentage of treatment successes (64% and 61% vs 15%; P<.001), and reduction in overall disease severity (30% and 29% vs 3%; P<.001). Local adverse effects did not differ significantly among the 3 groups after week 4. CONCLUSION: Short-contact 0.1% tazarotene gel therapy is a safe and effective new method of acne treatment. PMID- 11939811 TI - Improved differentiation of benign and malignant lymphadenopathy in patients with cutaneous melanoma by contrast-enhanced color Doppler sonography. AB - OBJECTIVE: To evaluate whether administration of a D-galactose-based signal enhancer is useful in color Doppler sonography (CDS) for better detection of vascularity patterns, which may help to differentiate malignant from benign lymph nodes in patients with cutaneous melanomas. DESIGN: Comparison of B-mode sonography, native CDS, and signal-enhanced CDS. SETTING: Department of Dermatology and Allergology, Ludwig-Maximilians-University, Munich, Germany. PATIENTS: Twenty examinations in 19 patients (median age, 60 years; 10 men) who presented with echo-poor structures suggestive of lymphadenopathy in B-mode sonography during follow-up for cutaneous melanomas. INTERVENTIONS: Histopathologic and follow-up examinations; documentation by color prints. MAIN OUTCOME MEASURES: Frequency of detection and description of different lymph node vascularity patterns in signal-enhanced CDS. RESULTS: Signal-enhanced CDS revealed additional information about vascularization of lymph node metastases, reactive lymph nodes, hematomas, and seromas, which was helpful for the differential diagnosis in 15 of 20 examinations. For lymph node metastases, signal enhancement facilitated the detection of accessory peripheral vessels in most investigations. Concerning reactive lymph nodes, hilar vessels in part with branching to the lymph node periphery could be identified only after application of the contrast enhancer in most patients. Quantitative variables could not be measured in all cases and did not help to differentiate between malignant and reactive lymph nodes. CONCLUSIONS: Administration of a D-galactose-based signal enhancer for CDS in patients with cutaneous melanomas can help to differentiate malignant from reactive lymph nodes, hematomas, or seromas. However, these promising results require confirmation in a prospective multicenter study. PMID- 11939812 TI - Is the loose anagen hair syndrome a keratin disorder? A clinical and molecular study. AB - OBJECTIVES: To report the clinical features of the loose anagen hair syndrome and to test the hypothesis that the typical gap between the hair and the inner root sheath may result from hereditary defects in the inner root sheath or the apposed companion layer. DESIGN: Case series. SETTING: A pediatric dermatology unit (referral center). PATIENTS: A consecutive sample of 17 children (13 girls). For 9 of them and their first-degree relatives, molecular analyses were performed in the K6HF gene with 50 appropriate controls. INTERVENTION: Minoxidil therapy (5% lotion) in 11 patients for 1 to 12 months. MAIN OUTCOME MEASURES: Clinical and follow-up features and determination of mutations in the K6HF gene. RESULTS: Most patients had easily pluckable hair with no sign of scalp inflammation or scarring. Ten patients seldom cut their hair, and 4 had unmanageable hair. One patient had hypodontia. Two patients had an additional clinical phenotype of diffuse partial woolly hair. The family history was positive for loose anagen hair syndrome in 5 patients. Marked improvement was noted after treatment with 5% minoxidil lotion in 7 of the 11 patients treated. Polymerase chain reaction analysis of the gene segments encoding the alpha-helical 1A and 2B subdomains of K6hf, the type II cytokeratin exclusively expressed in the companion layer, was performed in 9 families. In 3 of these 9 families, a heterozygous glutamic acid and lysine mutation, E337K, was identified in the L2 linker region of K6HF. CONCLUSIONS: Diffuse partial woolly hair can be associated with loose anagen hair syndrome. A keratin mutation, E337K in K6HF, was possibly causative in 3 of the 9 families studied. Another keratin, and possibly the type I partner of K6hf, could be responsible for loose anagen hair syndrome in other patients, or the gene involved may be a minor gene. PMID- 11939813 TI - Pain associated with injection of botulinum A exotoxin reconstituted using isotonic sodium chloride with and without preservative: a double-blind, randomized controlled trial. AB - CONTEXT: Botulinum A exotoxin is used for various indications, including the treatment of dynamic forehead lines. OBJECTIVE: To determine whether injection with botulinum A exotoxin reconstituted with preservative-containing normal saline (isotonic sodium chloride) is less painful than injection with exotoxin that has been reconstituted with preservative-free saline. DESIGN: Two arms: (1) retrospective study; (2) double-blind, randomized controlled trial. SETTING: A multiple-physician dermatology practice. PATIENTS: (1) Retrospective study-20 consecutive adult patients presenting for treatment of upper-face dynamic lines; (2) prospective study-15 consecutive adult patients presenting for treatment of upper-face dynamic lines. INTERVENTION: In prospective study only, one side (left or right) of the face was treated with exotoxin reconstituted with preservative containing saline, and the other side, with exotoxin reconstituted with preservative-free saline. MAIN OUTCOME MEASURES: (1) Retrospective study discomfort at current treatment (with preservative-containing saline) compared with discomfort with most recent prior treatment (with preservative-free saline); (2) prospective study-discomfort on the side treated with preservative-containing saline compared with discomfort on the side treated with preservative-free saline. RESULTS: (1) Retrospective study-18 (90%) of 20 patients reported that treatment with exotoxin reconstituted with preserved saline was less painful than prior treatment with exotoxin reconstituted with preservative-free saline; (2) prospective study-15 (100%) of 15 patients reported less pain in the side of their face treated with exotoxin reconstituted with preservative-containing saline (P<.001). Pain on the preservative-containing side was 54% less. No difference in treatment efficacy between the sides was observed by investigators or patients. CONCLUSION: Use of preservative-containing saline to reconstitute botulinum A exotoxin can significantly decrease patient discomfort on injection. PMID- 11939814 TI - Physician appearance is a matter of trust and responsibility. PMID- 11939815 TI - Loose anagen hair syndrome and loose anagen hair. PMID- 11939816 TI - Groin eruption in an HIV-positive man: epidermodysplasia verruciformis (EDV). PMID- 11939817 TI - Chronic erythematous desquamative plaques of the eyelids: discoid lupus erythematosis (DLE). PMID- 11939818 TI - Cutaneous eruption limited to skin covered by a swimming suit: mycosis fungoides. PMID- 11939819 TI - Alopecia, hypohidrosis, and ulcerations in a man: mycosis fungoides. PMID- 11939820 TI - Direct medical cost for surgical and medical treatment of condylomata acuminata. PMID- 11939822 TI - Imiquimod and genital herpes. PMID- 11939824 TI - Laser treatment of nevomelanocytic nevi: can results from an Asian study be applicable to the white population? PMID- 11939825 TI - Onset of non-AIDS Kaposi sarcoma during therapy with interferon alfa-2a in an 82 year-old man with concomitant cutaneous T-cell lymphoma. PMID- 11939826 TI - Metastatic silicone granuloma: lupus miliaris disseminatus faciei-like facial nodules and sicca complex in a silicone breast implant recipient. PMID- 11939827 TI - Delusional hyperhidrosis as a risk for medical overtreatment: a case of botulinophilia. PMID- 11939828 TI - Definition of axillary hyperhidrosis by gravimetric assessment. PMID- 11939829 TI - Endocrinologic changes in male patients with melanoma during interferon alfa-2b therapy. PMID- 11939830 TI - Topical 0.1% alitretinoin gel for classic Kaposi sarcoma. PMID- 11939831 TI - Unilateral gynecomastia induced by treatment with 1 mg of oral finasteride. PMID- 11939832 TI - American board of dermatology certification examination: preparation and perceptions. PMID- 11939833 TI - Sarcoidosis during combined interferon alfa and ribavirin therapy in 2 patients with chronic hepatitis C. PMID- 11939834 TI - Treatment of medium-brown solar lentigines using an alexandrite laser designed for hair reduction. PMID- 11939838 TI - A piece of my mind. Cord. PMID- 11939840 TI - Decade of work shows depression is physical. PMID- 11939841 TI - Future bright for light as dermatologic tool. PMID- 11939842 TI - As Americans age, geriatricians go missing. PMID- 11939847 TI - Rofecoxib vs celecoxib vs acetaminophen for treatment of osteoarthritis. PMID- 11939848 TI - Rofecoxib vs celecoxib vs acetaminophen for treatment of osteoarthritis. PMID- 11939850 TI - Acupuncture for the treatment of cocaine addiction. PMID- 11939851 TI - Acupuncture for the treatment of cocaine addiction. PMID- 11939852 TI - Acupuncture for the treatment of cocaine addiction. PMID- 11939853 TI - Acupuncture for the treatment of cocaine addiction. PMID- 11939854 TI - Acupuncture for the treatment of cocaine addiction. PMID- 11939856 TI - Current resident work hours: too many or not enough? PMID- 11939857 TI - Current resident work hours: too many or not enough? PMID- 11939859 TI - Changes in the treatment of depression in the United States: 1987-1997. PMID- 11939860 TI - Changes in the treatment of depression in the United States: 1987-1997. PMID- 11939862 TI - Trends in medication use in the United States. PMID- 11939864 TI - Long-term trends in mortality in the intensive care unit. PMID- 11939866 TI - Effect of Hypericum perforatum (St John's wort) in major depressive disorder: a randomized controlled trial. AB - CONTEXT: Extracts of Hypericum perforatum (St John's wort) are widely used for the treatment of depression of varying severity. Their efficacy in major depressive disorder, however, has not been conclusively demonstrated. OBJECTIVE: To test the efficacy and safety of a well-characterized H perforatum extract (LI 160) in major depressive disorder. DESIGN AND SETTING: Double-blind, randomized, placebo-controlled trial conducted in 12 academic and community psychiatric research clinics in the United States. PARTICIPANTS: Adult outpatients (n = 340) recruited between December 1998 and June 2000 with major depression and a baseline total score on the Hamilton Depression Scale (HAM-D) of at least 20. INTERVENTIONS: Patients were randomly assigned to receive H perforatum, placebo, or sertraline (as an active comparator) for 8 weeks. Based on clinical response, the daily dose of H perforatum could range from 900 to 1500 mg and that of sertraline from 50 to 100 mg. Responders at week 8 could continue blinded treatment for another 18 weeks. MAIN OUTCOME MEASURES: Change in the HAM-D total score from baseline to 8 weeks; rates of full response, determined by the HAM-D and Clinical Global Impressions (CGI) scores. RESULTS: On the 2 primary outcome measures, neither sertraline nor H perforatum was significantly different from placebo. The random regression parameter estimate for mean (SE) change in HAM-D total score from baseline to week 8 (with a greater decline indicating more improvement) was -9.20 (0.67) (95% confidence interval [CI], -10.51 to -7.89) for placebo vs -8.68 (0.68) (95% CI, -10.01 to -7.35) for H perforatum (P =.59) and 10.53 (0.72) (95% CI, -11.94 to -9.12) for sertraline (P =.18). Full response occurred in 31.9% of the placebo-treated patients vs 23.9% of the H perforatum treated patients (P =.21) and 24.8% of sertraline-treated patients (P =.26). Sertraline was better than placebo on the CGI improvement scale (P =.02), which was a secondary measure in this study. Adverse-effect profiles for H perforatum and sertraline differed relative to placebo. CONCLUSION: This study fails to support the efficacy of H perforatum in moderately severe major depression. The result may be due to low assay sensitivity of the trial, but the complete absence of trends suggestive of efficacy for H perforatum is noteworthy. PMID- 11939867 TI - Fish and omega-3 fatty acid intake and risk of coronary heart disease in women. AB - CONTEXT: Higher consumption of fish and omega-3 fatty acids has been associated with a lower risk of coronary heart disease (CHD) in men, but limited data are available regarding women. OBJECTIVE: To examine the association between fish and long-chain omega-3 fatty acid consumption and risk of CHD in women. DESIGN, SETTING, AND PARTICIPANTS: Dietary consumption and follow-up data from 84 688 female nurses enrolled in the Nurses' Health Study, aged 34 to 59 years and free from cardiovascular disease and cancer at baseline in 1980, were compared from validated questionnaires completed in 1980, 1984, 1986, 1990, and 1994. MAIN OUTCOME MEASURES: Incident nonfatal myocardial infarction and CHD deaths. RESULTS: During 16 years of follow-up, there were 1513 incident cases of CHD (484 CHD deaths and 1029 nonfatal myocardial infarctions). Compared with women who rarely ate fish (<1 per month), those with a higher intake of fish had a lower risk of CHD. After adjustment for age, smoking, and other cardiovascular risk factors, the multivariable relative risks (RRs) of CHD were 0.79 (95% confidence interval [CI], 0.64-0.97) for fish consumption 1 to 3 times per month, 0.71 (95% CI, 0.58-0.87) for once per week, 0.69 (95% CI, 0.55-0.88) for 2 to 4 times per week, and 0.66 (95% CI, 0.50-0.89) for 5 or more times per week (P for trend =.001). Similarly, women with a higher intake of omega-3 fatty acids had a lower risk of CHD, with multivariable RRs of 1.0, 0.93, 0.78, 0.68, and 0.67 (P<.001 for trend) across quintiles of intake. For fish intake and omega-3 fatty acids, the inverse association appeared to be stronger for CHD deaths (multivariate RR for fish consumption 5 times per week, 0.55 [95% CI, 0.33-0.90] for CHD deaths vs 0.73 [0.51-1.04]) than for nonfatal myocardial infarction. CONCLUSION: Among women, higher consumption of fish and omega-3 fatty acids is associated with a lower risk of CHD, particularly CHD deaths. PMID- 11939868 TI - Outcomes at 3 months after planned cesarean vs planned vaginal delivery for breech presentation at term: the international randomized Term Breech Trial. AB - CONTEXT: The Term Breech Trial found a significant reduction in adverse perinatal outcomes without an increased risk of immediate maternal morbidity with planned cesarean delivery compared with planned vaginal birth. No randomized controlled trial of planned cesarean delivery has measured benefits and risks of postpartum outcomes months after the birth. OBJECTIVE: To compare maternal outcomes of planned cesarean delivery and planned vaginal birth at 3 months post partum. DESIGN: Follow-up study to the Term Breech Trial, a randomized controlled trial conducted between January 9, 1997, and April 21, 2000. SETTING AND PARTICIPANTS: A total of 1596 of 1940 women from 110 centers worldwide who had a singleton fetus in breech presentation at term responded to a follow-up questionnaire at 3 months post partum. MAIN OUTCOME MEASURES: Breastfeeding; infant health; ease of caring for infant and adjusting to being a new mother; sexual relations and relationship with husband/partner; pain; urinary, flatal, and fecal incontinence; depression; and views regarding childbirth experience and study participation. RESULTS: Baseline information was similar for both the cesarean and vaginal delivery groups. Women in the planned cesarean delivery group were less likely to report urinary incontinence than those in the planned vaginal birth group (36/798 [4.5%] vs 58/797 [7.3%]; relative risk, 0.62; 95% confidence interval, 0.41 0.93). Incontinence of flatus was not different between groups but was less of a problem in the planned cesarean delivery group when it occurred (P =.006). There were no differences between groups in other outcomes. CONCLUSIONS: Planned cesarean delivery for pregnancies with breech presentation at term may result in a lower risk of incontinence and is not associated with an increased risk of other problems for women at 3 months post partum, although the effect on longer term outcomes is uncertain. PMID- 11939869 TI - Childhood cancer survivors' knowledge about their past diagnosis and treatment: Childhood Cancer Survivor Study. AB - CONTEXT: Adult survivors of childhood cancer are at risk for adverse effects later in life but may have limited access to information about their diagnosis and treatment. This knowledge is necessary to motivate them to seek medical follow-up and to report essential history to health care professionals. OBJECTIVE: To assess knowledge of adult survivors of childhood cancer about their primary cancer diagnosis and associated therapies. DESIGN, SETTING, AND PARTICIPANTS: Cross-sectional survey of 635 consecutive survivors (approximately 5%) drawn from 12 156 participants 18 years or older participating in the Childhood Cancer Survivor Study (a multiinstitutional cohort of individuals diagnosed between January 1, 1970, and December 31,1986, at an age <21 years, who had survived 5 years from diagnosis). The survey assessed knowledge of their cancer diagnosis and associated therapies in a 3- to 5-minute telephone questionnaire. MAIN OUTCOME MEASURES: Responses were compared with medical record data for accuracy, sensitivity, specificity, and positive and negative predictive value. RESULTS: Overall, 72% accurately reported their diagnosis with precision and 19% were accurate but not precise. Individuals with central nervous system (CNS) cancer (odds ratio, 5.1; 95% confidence interval, 2.6-9.9) and neuroblastoma (OR, 4.2; 95% CI, 1.8-9.6) were more likely not to know their cancer diagnosis. Participants' accuracy rates for reporting their treatment history was 94% for chemotherapy, 89% for radiation, and 93% for splenectomy. Among those who received anthracyclines, only 30% recalled receiving daunorubicin therapy and 52% recalled receiving doxorubicin therapy, even after prompting with the drugs' names. Among those who received radiotherapy, 70% recalled the site of radiotherapy. History of receiving a written medical summary, attending a long term follow-up clinic, and anxiety about late effects were not associated with greater knowledge. CONCLUSIONS: Important knowledge deficits exist among adult survivors of childhood cancer regarding basic aspects of their diagnosis and treatment. Such deficits could impair survivors' ability to seek and receive appropriate long-term follow-up care. PMID- 11939870 TI - Placebo response in studies of major depression: variable, substantial, and growing. AB - CONTEXT: Intense debate persists about the need for placebo-controlled groups in clinical trials of medications for major depressive disorder (MDD). There is continuing interest in the development of new medications, but because effective antidepressants are already available, ethical concerns have been raised about the need for placebo groups in new trials. OBJECTIVE: To determine whether the characteristics of placebo control groups in antidepressant trials have changed over time. DATA SOURCES AND STUDY SELECTION: We searched MEDLINE and PsychLit for all controlled trials published in English between January 1981 and December 2000 in which adult outpatients with MDD were randomly assigned to receive medication or placebo. Seventy-five trials met our criteria for inclusion. DATA EXTRACTION: Data were extracted from the articles by 2 of the authors and discrepancies were resolved via discussion and additional review by a third author. DATA SYNTHESIS: The mean (SD) proportion of patients in the placebo group who responded was 29.7% (8.3%) (range, 12.5%-51.8%). Most studies examined more than a single active medication, and, in the active medication group with the greatest response, the mean (SD) proportion of patients responding was 50.1% (9.0%) (range, 31.6% 70.4%). Both the proportion of patients responding to placebo and the proportion responding to medication were significantly positively correlated with the year of publication (for placebo: n = 75; r = 0.45; 95% confidence interval [CI], 0.25 0.61; P<.001; for medication: n = 75; r = 0.26; 95% CI, 0.03-0.46; P =.02). The association between year of publication and response rate was more statistically robust for placebo than medication. CONCLUSIONS: The response to placebo in published trials of antidepressant medication for MDD is highly variable and often substantial and has increased significantly in recent years, as has the response to medication. These observations support the view that the inclusion of a placebo group has major scientific importance in trials of new antidepressant medications and indicate that efforts should continue to minimize the risks of such studies so that they may be conducted in an ethically acceptable manner. PMID- 11939871 TI - Device therapy for cardiac arrhythmias. PMID- 11939873 TI - JAMA patient page. Cancer and children. PMID- 11939872 TI - Placebo in clinical trials for depression: complexity and necessity. PMID- 11939874 TI - Psychiatry and stigmatization. PMID- 11939875 TI - Overcoming pessimism about treatment of addiction. PMID- 11939876 TI - Misperceptions behind mental health policy. PMID- 11939877 TI - Declining student interest in psychiatry. PMID- 11939878 TI - Chlorpromazine and the psychopharmacologic revolution. PMID- 11939884 TI - Aging in the 21st century: a call for papers. PMID- 11939885 TI - Chronic idiopathic axonal polyneuropathy and successful aging of the peripheral nervous system. PMID- 11939886 TI - Cerebrotendinous xanthomatosis: a rare disease with diverse manifestations. AB - This mini-review deals with a new appraisal of cerebrotendinous xanthomatosis. In addition to neurologic symptoms, patients with cerebrotendinous xanthomatosis develop cataracts, diarrhea, Achilles tendon xanthoma, atherosclerotic vascular disease, and many other abnormalities. Although the pathophysiology of the disease is not completely understood, excess production and consequent accumulation of cholestanol in tissues may play a crucial role. Chenodeoxycholic acid is the most effective therapy. The causative role and detrimental effects (at a low plasma level) of cholestanol merit further investigation. PMID- 11939887 TI - Chronic idiopathic axonal polyneuropathy and successful aging of the peripheral nervous system in elderly people. AB - BACKGROUND: Chronic idiopathic axonal polyneuropathy (CIAP) is a frequent neurologic disorder in elderly persons. In view of the aging population, it is important to know the long-term prognosis of CIAP. OBJECTIVES: To determine if CIAP is influenced by the superposition of the effects of aging and to evaluate the severity of CIAP according to the disease duration. DESIGN: Controlled cohort study. SETTING: Outpatient clinic for neuromuscular diseases at the University Medical Center Utrecht, Utrecht, the Netherlands. PARTICIPANTS AND METHODS: One hundred twenty-seven patients with CIAP and 108 age-matched control subjects were included. We defined CIAP on the basis of symmetrical distal sensory or sensorimotor symptoms and signs with evolution over at least 6 months, exclusion of causes by history taking, results of clinical and laboratory investigations, and electrophysiologic findings that agreed with the diagnosis of axonal polyneuropathy. RESULTS: No important neurologic or electrophysiologic differences were found between patients with early-onset (before the age of 65 years) and late-onset (at or after the age of 65 years) CIAP, but patients with early-onset CIAP who had a short disease duration (<10 years) experienced more disability than patients with late-onset CIAP who had a similar disease duration. Old controls (age of 65 years or older) more often had symptoms, sensory signs in the legs, absent ankle jerks, and lower mean distal amplitudes of compound muscle action potentials and sensory nerve action potentials than young controls (aged <65 years). Absence of the sural nerve sensory nerve action potentials or presence of spontaneous muscle fiber activity in the anterior tibial muscle was common in patients with CIAP (51% and 60%, respectively), but exceptional (both 2%) in controls. CONCLUSIONS: Neither aging of the peripheral nervous system nor disease duration affects CIAP to a considerable degree, but CIAP has a greater influence on the daily life of nonretired patients with early-onset CIAP. The diagnosis of axonal polyneuropathy is probably supported best by either the absence of the sural nerve sensory nerve action potentials or the presence of spontaneous muscle fiber activity in the anterior tibial muscle. PMID- 11939888 TI - Incidence of acute femoral neuropathy following renal transplantation. AB - BACKGROUND: Case reports exist of femoral neuropathy following renal transplantation (RTSP) with possible pathophysiology, including direct compression and nerve ischemia. However, the occurrence of acute femoral neuropathy (AFN) following RTSP has not been studied prospectively. OBJECTIVE: To determine the occurrence of AFN following RTSP. METHODS: We prospectively studied the occurrence of AFN following RTSP from June 1, 1998, to October 31, 1999. A total of 184 RTSPs were performed during this period. All the patients had end stage renal failure and had effective hemodialysis before RTSP. All patients with AFN underwent neurologic examination, nerve conduction and electromyographic studies (5 to 7 days after the onset of symptoms), and magnetic resonance imaging or computed tomography of pelvis and lumbosacral spine within 24 hours of onset of symptoms. RESULTS: Four (2.2%) of 184 patients developed AFN (ipsilateral to the RTSP surgery) postoperatively between 24 (3 patients) and 48 hours. All the patients achieved good renal function after RTSP. All the patients had excellent recovery of motor function in 4 to 9 months. CONCLUSION: We believe that AFN following RTSP is an uncommon (2.2%) complication from which patients have an excellent chance of recovery. PMID- 11939889 TI - Thrombolysis in patients with acute stroke caused by cervical artery dissection: analysis of 9 patients and review of the literature. AB - BACKGROUND: Results of recently published studies suggest that intravenous thrombolysis (IVT) and local intra-arterial thrombolysis (LIT) are feasible procedures in acute stroke after cervical artery dissection (CAD). OBJECTIVES: To describe 9 patients with acute stroke caused by CAD who were treated by LIT (n = 7) or IVT (n = 2) and to review the literature. METHODS: Retrospective analysis of clinical and neuroradiological findings; literature review from 1980 to present. MAIN OUTCOME MEASURE: Modified Rankin Scale (mRS) score. RESULTS: Of 7 patients treated with LIT, 3 had good outcomes (mRS score of 0-2) and 4 had bad outcomes (mRS score of 3-6) at 3 months. The 2 patients who had received IVT recovered to mRS scores of 0 and 3. Twenty-one patients were identified in the literature. Overall (N = 30), in the IVT group (n = 19), the outcome was good in 8 patients (42%) and bad in 11 (58%); in the LIT group (n = 11), 6 patients (55%) had a good outcome and 5 (45%) had a bad outcome. Overall, 47% (14/30) of the patients (IVT and LIT groups) had a good outcome. Total mortality was 13% (4/30). There were no secondary complications due to extension of wall hematoma or angiography. One symptomatic hemorrhage occurred. CONCLUSIONS: Thrombolysis is feasible in acute stroke caused by CAD. Local complications from extension of wall hematoma did not occur. Further prospective studies are needed to determine the safety and efficacy of thrombolysis in the special circumstance of acute stroke caused by CAD. PMID- 11939890 TI - Occlusion due to carotid artery dissection: a more severe disease than previously suggested. AB - BACKGROUND: Stroke due to internal carotid artery dissection is considered to have a good prognosis. OBJECTIVE: To determine whether the prognosis of internal carotid artery dissection is worse than classically reported by comparing the characteristics of patients who had an acute ischemic stroke admitted to a population-based primary care center with internal carotid artery occlusion due to either dissection (DO) or atherothrombosis (AO). PATIENTS AND METHODS: Among 3502 patients admitted to our population-based primary care center, DO (n = 73) was diagnosed by angiography or magnetic resonance imaging, while AO (n = 81) was diagnosed by angiography. The characteristics of patients with DO or AO were compared using univariate and multivariate analysis. RESULTS: Patients with DO were younger (mean [SD] age, 44.6 [10] vs. 60.1 [10] years, P<.001), had fewer vascular risk factors, and presented more frequently with global middle cerebral artery territory involvement (42% vs. 17%, P<.05) and less frequently with watershed infarcts (3% vs. 19%, P<.05) than patients with AO. Unexpectedly, patients with DO were noted to have more severe clinical impairment, with an increased frequency of decreased consciousness, and a poorer outcome at 1 month. Multivariate analysis showed that the independent factors associated with DO were age younger than 55 years, nonsmoker, no history of hypertension, headache at presentation, and global aphasia. CONCLUSIONS: Patients with DO are younger and are initially seen with fewer risk factors than patients with AO, but their clinical features and prognosis are worse. Large infarcts involving the whole middle cerebral artery territory that may be due to the lack of collateral circulation are responsible for the bad prognosis of patients with DO. PMID- 11939891 TI - Stroke or transient ischemic attacks with basilar artery stenosis or occlusion: clinical patterns and outcome. AB - BACKGROUND: Basilar artery occlusion (BAO) is associated with a high mortality rate, although cases with spontaneous favorable outcomes have recently been reported, and basilar artery stenosis (BAS) has received little consideration until now. OBJECTIVE: To study the prognostic clinical factors by testing numerous combinations of admission status characteristics of patients with brain ischemia caused by BAO or BAS. METHODS: We conducted a retrospective review from the Lausanne Stroke Registry (group 1) of patients with stroke or transient ischemic attack caused by BAS less than 50% or BAO as diagnosed by magnetic resonance angiography who were not treated by thrombolysis. Neurologic findings on admission were correlated with outcomes. We compared clinical patterns associated with poor outcomes in group 1 with those in patients with stroke who died from BAO or BAS (confirmed at autopsy) (group 2). RESULTS: Eighty-eight patients were studied. The outcomes of patients with stroke in group 1 (35/43) was poor (severe disability or death) in 54% of cases. A statistical analysis revealed that 4 factors-dysarthria, pupillary disorders, lower cranial nerve involvement, and consciousness disorders on admission-were strongly (P<.001) associated with poor outcomes. The multivariate analysis showed that the outcome was poor in 100% of cases in which consciousness disorders or the combination of the remaining 3 factors were present, whereas in the absence of these factors, a poor outcome was reported in only 11%. In 87% of the 45 patients with stroke in group 2, the same clinical patterns were present on admission. CONCLUSIONS: The prognosis of BAS greater than 50% or BAO is diverse and certain clinical characteristics seem to predict a lower risk of poor outcome. Their presence may help to decide the most suitable therapy. PMID- 11939892 TI - Dopaminergic function and dopamine transporter binding assessed with positron emission tomography in Parkinson disease. AB - BACKGROUND: Measuring progression of Parkinson disease (PD) using positron emission tomography may help demonstrate the efficacy of neuroprotective treatments. To date, (18)F-dopa has been the gold standard to measure presynaptic dopaminergic function in PD, but this tracer might overestimate the rate of neuronal death in PD because its uptake also depends on dopamine turnover rather than exclusively on the density of dopaminergic terminals in the striatum. The latter might be assessed using newly developed ligands of the membrane dopamine transporter. OBJECTIVE: To compare the striatal uptakes of (18)F-dopa and (76)Br FE-CBT, a dopamine transporter ligand, in patients with PD. PATIENTS AND METHODS: The striatal uptakes of (76)Br-FE-CBT and (18)F-dopa were compared using positron emission tomography in 10 patients with early PD and 8 with advanced PD. Correlation of uptakes with motor performance was investigated. RESULTS: The reduction in (76)Br-FE-CBT binding to 43% of control values was more severe than the reduction in (18)F-dopa uptake (63% of control values) in the putamen of patients with early PD. No significant difference was found between either tracer's uptake in the putamen of patients with advanced PD. Motor performance was highly correlated to (18)F-dopa uptake, whereas correlation to (76)Br-FE-CBT binding was weak. CONCLUSIONS: Uptake of (18)F-dopa may be up-regulated in early PD, suggesting a compensatory increase of dopamine synthesis in surviving dopaminergic terminals. Positron emission tomography dopamine transporter ligands and (18)F-dopa give complementary information on the presynaptic status of the nigrostriatal dopaminergic system and might be associated to investigate the efficacy of neuroprotective treatments in PD. PMID- 11939893 TI - Effect of extrapyramidal signs and Lewy bodies on survival in patients with Alzheimer disease. AB - BACKGROUND: Patients with Alzheimer disease (AD) who have psychiatric and parkinsonian symptoms experience faster cognitive deterioration and shorter survival vs those without such disease features. Extrapyramidal signs (EPSs) in particular have been associated with the presence of Lewy bodies (LBs) on autopsy and with poorer survival in patients with AD. Lewy bodies found at autopsy are not always correlated with EPSs during late life. OBJECTIVE: To determine whether the association between LBs and age at death is modified by the presence of EPSs, hallucinations, or delusions. DESIGN: An autopsy series of patients with clinically diagnosed AD. SETTINGS: Three AD clinics (San Diego and Sacramento, Calif, and Portland, Ore). PATIENTS: Data on 379 patients with a clinical diagnosis of AD who were initially assessed between May 1, 1984, and August 1, 1996, and who were autopsied between January 1, 1990, and April 1, 1998, were pooled from 3 AD centers. MAIN OUTCOME MEASURES: Presence of LBs on autopsy and differences in age at death in those with EPSs, LBs, or both. RESULTS: Individuals with EPSs at initial assessment were more than 3 times as likely to have LBs at autopsy than were those without EPSs. Age at death was younger in those with LBs and EPSs than in those with LBs only and those without EPSs or LBs. CONCLUSIONS: The presence of EPSs in patients with AD indicates worse prognosis and may be related to underlying LBs. The presence of EPSs is a strong predictor of LBs. PMID- 11939894 TI - Association between apolipoprotein E genotype and Alzheimer disease in African American subjects. AB - BACKGROUND: The association between Alzheimer disease (AD) and genotypes at the apolipoprotein E (APOE) locus has been confirmed in numerous populations worldwide, but appears to be inconsistent in African American subjects. OBJECTIVE: To investigate the association between APOE genotypes and AD in elderly African American subjects. DESIGN: Clinic-based, multicenter case-control study and a family study. PARTICIPANTS: A total of 338 African American probands meeting criteria for probable or definite AD, 301 cognitively healthy, elderly unrelated control subjects (spouses and community volunteers), and 108 siblings of 88 AD probands. MAIN OUTCOME MEASURES: Odds of AD according to APOE genotype. RESULTS: Compared with individuals with the APOEepsilon3/epsilon3, the odds of having AD were significantly increased among those with 1 or more copies of the epsilon4 allele; the odds ratio (OR) for the epsilon3/epsilon4 genotype was 2.6 (95% confidence interval [CI], 1.8-3.7), and the OR for the epsilon4/epsilon4 genotype was 10.5 (95% CI, 5.1-21.8). These risks decreased substantially after 68 years of age. The risk for AD was lower among individuals with the epsilon2/epsilon3 genotype (OR, 0.41; 95% CI, 0.22-0.79). The patterns of association were similar in men and women. These results obtained from comparisons of unrelated AD patients and controls were bolstered by results of analysis of family data that showed preferential transmission of the epsilon4 allele to demented siblings (P<<.001) and of the epsilon2 allele to nondemented siblings (P=.005). CONCLUSIONS: The presence of 1 or 2 epsilon4 alleles is a determinant of AD risk in African American subjects. The age-related risk for decline associated with the epsilon4 allele and the apparent protective effect of the epsilon2 allele are similar to patterns observed in white subjects. PMID- 11939895 TI - Independent predictors of cognitive decline in healthy elderly persons. AB - BACKGROUND: Several studies have shown that individually memory, hippocampal volume, and motor measures presage the onset of dementia. It is unclear if these independently contribute to the prediction of mild cognitive impairment. OBJECTIVE: To determine the ability of memory, hippocampal volume, and a gait speed to independently predict cognitive decline in healthy elderly persons. DESIGN: A prospective, longitudinal, observational cohort study with a mean follow-up of 6 years. PARTICIPANTS: One hundred eight optimally healthy elderly cognitively intact subjects. MAIN OUTCOME MEASURES: Any cognitive impairment noted on the Clinical Dementia Rating Scale (score = 0.5) or persistent or progressive cognitive impairment. Cox modeling determined if time to onset of cognitive impairment was associated with baseline logical memory II test score (a measure of delayed recall), hippocampal volume (magnetic resonance imaging), or gait speed (time to walk 30 ft [9 m]) independent of age, sex, depression, or the allele producing the epsilon4 type of apolipoprotein E (APOE epsilon4). RESULTS: Questionable dementia occurred in 48 participants in a mean (SD) of 3.7 (2.4) years. This progressed to persistent cognitive impairment in 38 of these participants in a mean (SD) of 4.4 (2.4) years. Logical memory II test performance and hippocampal volume each predicted onset of questionable dementia, independent of age and sex. Time to walk 30 ft additionally contributed independently to the prediction of time to onset of persistent cognitive impairment. Possessing the APOE epsilon4 allele and depression did not enter either model significantly. CONCLUSIONS: Models combining multiple risk factors should refine the prediction of questionable dementia and persistent cognitive impairment, harbingers of dementia. Individuals at risk for cognitive impairment may represent a high-risk group for intervention. PMID- 11939896 TI - Differences in tau and apolipoprotein E polymorphism frequencies in sporadic frontotemporal lobar degeneration syndromes. AB - BACKGROUND: Frontotemporal lobar degeneration (FTLD) has different clinical phenotypes and is associated with several pathologic findings, most commonly dementia lacking distinctive histology or Pick disease. We know that the tau H1 haplotype is associated with some clinical and histologic phenotypes, for example, progressive supranuclear palsy and corticobasal degeneration. Furthermore, the apolipoprotein epsilon4 allele (APOE epsilon4) may be associated with Pick disease. OBJECTIVE: To determine if different clinical phenotypes of FTLD are associated with different tau haplotype and APOE allele frequencies. PATIENTS AND METHODS: All patients with FTLD with available DNA specimens (n = 63) seen at the Mayo Clinic, Jacksonville, Fla, were retrospectively classified according to the following clinical phenotypes: frontal dementia (FD); progressive, nonfluent aphasia (PA); or fluent, anomic aphasia (AA). DNA specimens were genotyped for APOEallele and tau haplotype frequencies and were compared with cognitively normal patients (n = 338) and patients with Alzheimer disease (AD) (n = 193). RESULTS: Patients with AA had increased APOE epsilon4 frequency (30.4%) compared with patients with FD (14.8%, P=.04) and cognitively normal patients (11.1%, P<.001). Patients with AA also had increased tau H2 haplotype (37.0%) frequency compared with patients with FD (11.1%,P=.002), patients with AD (21.8%, P=.02), and cognitively normal patients (19.8%, P=.004). The increase in tau H2 haplotype frequency (50.0%) is especially pronounced in patients with AA who are APOE epsilon4 positive compared with patients with FD (18.8%, P=.04), patients with AD (24.8%, P=.005), and cognitively normal patients (15.3%, P<.001).APOE epsilon4 and tau H2 haplotype frequencies are not significantly different in patients with FD and PA compared with healthy patients. CONCLUSIONS: Clinical subtypes of FTLD have different tau and APOE genotype frequencies, suggesting these genes may influence the clinical presentation. Further studies should be performed to confirm this finding and to see if the pathologic phenotypes are also associated with different tau and APOE genotype frequencies. PMID- 11939897 TI - Significance of planum temporale and planum parietale morphologic features in neurofibromatosis type 1. AB - BACKGROUND: Neurofibromatosis type 1 (NF-1) is associated with learning disabilities and cognitive impairment in childhood and adolescence. Individuals with NF-1 have a propensity for brain hyperintensities on T2-weighted magnetic resonance images, macrocephaly, and optic gliomas. Few clear relationships between these central nervous system abnormalities and cognitive function, however, have been found in this population. OBJECTIVES: To determine whether planum temporale (PT) and planum parietale (PP) morphologic features are associated with learning disabilities in NF-1. PATIENTS AND METHODS: We measured and compared the surface area, gray matter volume, and asymmetry of the PT and PP on T1-weighted MRIs from 24 children and adolescents with NF-1 and an equal number of controls. Relationships between these measurements and cognitive and academic achievement scores were examined. RESULTS: The left PT in boys with NF-1 was significantly smaller in both surface area and gray matter volume compared with girls with NF-1 and controls. Boys with NF-1 also showed greater symmetry between the left and right hemispheres in this region compared with girls with NF 1 and controls, who showed a pattern of left greater than right asymmetry of the PT. Intelligence-based discrepancy scores of reading and math achievement, which are commonly used to define learning disabilities, were significantly related to PT asymmetry in the NF-1 group as a whole. Less leftward asymmetry of the PT was associated with poorer reading and math achievement in relation to intellectual test scores. CONCLUSIONS: The high susceptibility of individuals with NF-1 to develop reading and other learning disabilities seems to be related to the development of the sylvian fissure. These results provide further support for the hypothesized association between sylvian fissure morphologic features and learning disabilities. PMID- 11939898 TI - Trinucleotide repeats in 202 families with ataxia: a small expanded (CAG)n allele at the SCA17 locus. AB - BACKGROUND: Ten neurodegenerative disorders characterized by spinocerebellar ataxia (SCA) are known to be caused by trinucleotide repeat (TNR) expansions. However, in some instances the molecular diagnosis is considered indeterminate because of the overlap between normal and affected allele ranges. In addition, the mechanism that generates expanded alleles is not completely understood. OBJECTIVE: To examine the clinical and molecular characteristics of a large group of Portuguese and Brazilian families with ataxia to improve knowledge of the molecular diagnosis of SCA. PATIENTS AND METHODS: We have (1) assessed repeat sizes at all known TNR loci implicated in SCA; (2) determined frequency distributions of normal alleles and expansions; and (3) looked at genotype phenotype correlations in 202 unrelated Portuguese and Brazilian patients with SCA. Molecular analysis of TNR expansions was performed using polymerase chain reaction amplification. RESULTS: Patients from 110 unrelated families with SCA showed TNR expansions at 1 of the loci studied. Dominantly transmitted cases had (CAG)(n) expansions at the Machado-Joseph disease gene (MJD1) (63%), at SCA2 (3%), the gene for dentatorubropallidoluysian atrophy (DRPLA) (2%), SCA6 (1%), or SCA7 (1%) loci, or (CTG)(n) expansions at the SCA8 (2%) gene, whereas (GAA)(n) expansions in the Freidreich ataxia gene (FRDA) were found in 64% of families with recessive ataxia. Isolated patients also had TNR expansions at the MJD1 (6%), SCA8 (6%), or FRDA (8%) genes; in addition, an expanded allele at the TATA binding protein gene (TBP), with 43 CAGs, was present in a patient with ataxia and mental deterioration. Associations between frequencies of SCA2 and SCA6 and a frequency of large normal alleles were found in Portuguese and Brazilian individuals, respectively. Interestingly, no association between the frequencies of DRPLA and large normal alleles was found in the Portuguese group. CONCLUSIONS: Our results show that (1) a significant number of isolated cases of ataxia are due to TNR expansions; (2) expanded DRPLA alleles in Portuguese families may have evolved from an ancestral haplotype; and (3) small (CAG)(n) expansions at the TBP gene may cause SCA17. PMID- 11939899 TI - Progressive dementia and hypersomnolence with dream-enacting behavior: oneiric dementia. AB - BACKGROUND: Sleep disorders are associated with several types of degenerative dementias, including Alzheimer and prion diseases. Animal models have demonstrated abolition of rapid eye movement atonia, resulting in dream-enacting complex movements termed oneiric behavior, and patients with fatal familial insomnia may have vivid dreams that intrude on wakefulness. OBJECTIVE: To describe a new form of progressive dementia with hypersomnia and oneiric behavior. METHODS: Neuropsychological and polysomnographic studies of a middle aged woman with a progressive dementia, excessive daytime sleepiness, and a vertical gaze palsy. RESULTS: Neuropsychological testing revealed decreased verbal fluency, impaired attention and working memory, amnesia, poor recall, and bradyphrenia with hypersomnia. Polysomnography revealed a rapid eye movement behavioral disorder with complete absence of slow wave sleep. Prion protein analysis did not reveal the mutation associated with fatal familial insomnia, and other diagnostic test findings were unrevealing. CONCLUSION: Our patient had a previously unreported syndrome of progressive dementia associated with rapid eye movement behavioral disorder and the absence of slow wave sleep. PMID- 11939900 TI - Orbital myositis posing as cluster headache. AB - OBJECTIVE: To describe the case of a patient with recurrent orbital myositis who was thought to have cluster headaches for 6 years. DESIGN AND SETTING: Case report in an outpatient neuro-ophthalmology clinic. PATIENT: A 24-year-old man developed unilateral supraorbital pain, lacrimation, conjunctival hyperemia, nasal congestion, proptosis, and painful eye movements. The pain intensity varied over the course of each day and disappeared after 1 month. He had multiple attacks responsive to prednisone that were separated by months over the ensuing 6 years. Neuroimaging revealed an enlarged extraocular muscle. CONCLUSIONS: Overlap in symptoms between recurrent orbital myositis and cluster headache delayed the diagnosis in this patient. Orbital myositis should be considered in patients with atypical cluster headache characterized by proptosis, painful eye movements, and pain that does not completely resolve after 3 hours. PMID- 11939902 TI - Intravascular lymphomatosis presenting as posterior leukoencephalopathy. PMID- 11939901 TI - Abnormal diffusion-weighted magnetic resonance imaging in Creutzfeldt-Jakob disease following corneal transplantations. AB - BACKGROUND: The value of magnetic resonance imaging of the brain in the diagnosis of iatrogenic cases of Creutzfeldt-Jakob disease has been questioned. OBJECTIVE: To illustrate the value of magnetic resonance imaging of the brain in the diagnosis of iatrogenic Creutzfeldt-Jakob disease. METHODS: Case report. RESULTS: A patient with a history of 3 corneal transplantations exhibited the alien hand sign on initial examination. Diffusion-weighted magnetic resonance imaging of the brain revealed prominent cortical diffusion abnormalities. During the following months, the patient developed rapidly progressive dementia. The diagnosis of Creutzfeldt-Jakob disease was proven by brain biopsy. CONCLUSION: Brain magnetic resonance imaging, particularly diffusion-weighted magnetic resonance imaging, can be very helpful in the diagnosis of Creutzfeldt-Jakob disease, even in suspected iatrogenic cases. PMID- 11939903 TI - Harvey Cushing and medulloblastoma. PMID- 11939904 TI - Neurological deficits in patients with celiac disease. PMID- 11939905 TI - Mechanism of negative regulation of rat glutathione S-transferase A2 by the cytokine interleukin 6. AB - A decrease in concentration of some liver proteins, including the detoxification enzyme glutathione S-transferase A2 (rGSTA2), occurs during the acute-phase response. Interleukin 6 (IL-6) with dexamethasone (DEX) decreases transcription of rGSTA2 in rat hepatocytes. The promoter region that mediates suppression of rGSTA2 was localized to 150 bp. These 150 bp were divided and used for electrophoretic mobility-shift assays. Induction of a protein that specifically bound to an oligonucleotide from this region required new protein synthesis and IL-6 with DEX in the culture media. The protein bound to part of the hepatocyte nuclear factor 1 (HNF1) site but was different from and did not displace HNF1. A core sequence, TGATT, was required for binding. The protein also bound to an HNF1 site in the albumin promoter. We hypothesize that IL-6 along with DEX induced a novel protein that decreased transcription of rGSTA2 and possibly albumin by interfering with the transactivating function of HNF1. The protein may be an important negative regulator of transcription during the acute-phase response. PMID- 11939906 TI - Differential metabolism of dihomo-gamma-linolenic acid and arachidonic acid by cyclo-oxygenase-1 and cyclo-oxygenase-2: implications for cellular synthesis of prostaglandin E1 and prostaglandin E2. AB - Prostaglandin (PG) E(1) has been shown to possess anti-inflammatory properties and to modulate vascular reactivity. These activities are sometimes distinct from those of PGE(2), suggesting that endogenously produced PGE(1) may have some beneficial therapeutic effects compared with PGE(2). Increasing the endogenous formation of PGE(1) requires optimization of two separate processes, namely, enrichment of cellular lipids with dihomo-gamma-linolenic acid (20:3 n-6; DGLA) and effective cyclo-oxygenase-dependent oxygenation of substrate DGLA relative to arachidonic acid (AA; 20:4 n-6). DGLA and AA had similar affinities (K(m) values) and maximal reaction rates (V(max)) for cyclo-oxygenase-2 (COX-2), whereas AA was metabolized preferentially by cyclo-oxygenase-1 (COX-1). To overcome the kinetic preference of COX-1 for AA, CP-24879, a mixed Delta(5)/Delta(6) desaturase inhibitor, was used to enhance preferential accumulation of DGLA over AA in cells cultured in the presence of precursor gamma-linolenic acid (18:3 n-6). This protocol was tested in two cell lines and both yielded a DGLA/AA ratio of approx. 2.8 in the total cellular lipids. From the enzyme kinetic data, it was calculated that this ratio should offset the preference of COX-1 for AA over DGLA. PGE(1) synthesis in the DGLA-enriched cells was increased concurrent with a decline in PGE(2) formation. Nevertheless, PGE(1) synthesis was still substantially lower than that of PGE(2). It appears that employing a dietary or a combined dietary/pharmacological paradigm to augment the cellular ratio of DGLA/AA is not an effective route to enhance endogenous synthesis of PGE(1) over PGE(2), at least in cells/tissues where COX-1 predominates over COX-2. PMID- 11939907 TI - Kinetics of inter-domain electron transfer in flavocytochrome cellobiose dehydrogenase from the white-rot fungus Phanerochaete chrysosporium. AB - The pre-steady-state kinetics of inter-domain electron transfer in the extracellular flavocytochrome cellobiose dehydrogenase from Phanerochaete chrysosporium was studied using various values of pH and substrate concentration. Monitoring at the isosbestic point of each prosthetic group indicated that the reductive half-reactions of flavin and haem were biphasic and monophasic respectively. When the observed rates of the flavin and haem reactions were plotted against substrate concentration, the behaviour of the second phase of the flavin reduction was almost identical with that of haem reduction at all substrate concentrations and pH values tested, suggesting that the formation of flavin semiquinone and haem reduction involve the same electron transfer reaction. Although flavin reduction by cellobiose was observed in the range of pH 3.0-7.0, the velocity of the next electron transfer step decreased with increase of pH and was almost zero above pH 6.0. The second phase of flavin reduction and the haem reduction were inhibited similarly by high concentrations of the substrate, whereas the first phase of flavin reduction showed a hyperbolic relation to the cellobiose concentration. Increase in pH enhanced the substrate inhibition of haem reduction but not the initial flavin reduction. Moreover, the dissociation constant K(d) of flavin reduction and the substrate inhibition constant K(i) of haem reduction decreased similarly with an increase of pH. From these results, it is evident that binding of cellobiose to the active site inhibits electron transfer from flavin to haem. PMID- 11939908 TI - Multiple regions within the promoter of the murine Ifnar-2 gene confer basal and inducible expression. AB - The (murine) type I interferon (IFN) receptor, muIfnar-2, is expressed ubiquitously, and exists as both transmembrane and soluble forms. In the present study we show that the gene encoding muIfnar-2 spans approx. 33 kb on mouse chromosome 16, and consists of nine exons and eight introns. The three mRNA splice variants resulting in one transmembrane (muIfnar-2c) and two soluble (muIfnar-2a/2a') mRNA isoforms are generated by alternative RNA processing of the muIfnar-2 gene. Treatment of a range of murine cell lines with a combination of type I and II IFN showed that the muIfnar-2a and -2c mRNA isoforms were up regulated independently of each other in L929 fibroblasts and hepa-1c1c7 hepatoma cells, but not in M1 myeloid leukaemia cells. Analysis of the 5' flanking region of muIfnar-2 using promoter-luciferase reporter constructs defined three regulatory regions: a region proximal to exon 1, conferring high basal expression, a distal region conferring inducible expression, and a negative regulatory region between the two. These data represent the first promoter analysis of a type I IFN receptor and, taken together with our previous data demonstrating high expression levels and dual biological functions for muIfnar-2a protein, suggests that the regulation of muIfnar-2 isoform expression may be an important way of modulating type I IFN responses. PMID- 11939910 TI - Victims of awareness. AB - BACKGROUND: Intraoperative awareness with explicit recall may be followed by long lasting mental symptoms. However, the average risk for developing mental sequelae after awareness, and the average severity and the duration of symptoms has not previously been illustrated in a consecutive series of awareness cases. METHODS: Nine patients among 18 consecutive, prospectively identified cases of intraoperative awareness with recall could be located after approximately 2 years and agreed to an interview about possible persisting problems. RESULTS: Four of the nine interviewed patients were still severely disabled due to psychiatric/psychological sequelae. All of these patients had experienced anxiety during the period of awareness, but only one had complained about pain. Another three patients had less severe, transient mental symptoms, although they could cope with these in daily life. Two patients denied any sequelae from their awareness episode. CONCLUSIONS: Up to 3 weeks after their unsuccessful anesthetic, repeated information and discussions had been offered. Despite the fact that all patients at that time claimed to be satisfied with this management, and eventually considered no further contacts necessary, this was obviously inaccurate. Therefore, professional psychiatric assessment, treatment and long term follow-up should constitute standard practice for all patients who have experienced intraoperative awareness. PMID- 11939909 TI - Association of brain gamma-tubulins with alpha beta-tubulin dimers. AB - gamma-Tubulin is necessary for nucleation and polar orientation of microtubules in vivo. The molecular mechanism of microtubule nucleation by gamma-tubulin and the regulation of this process are not fully understood. Here we show that there are two gamma-tubulin forms in the brain that are present in complexes of various sizes. Large complexes tend to dissociate in the presence of a high salt concentration. Both gamma-tubulins co-polymerized with tubulin dimers, and multiple gamma-tubulin bands were identified in microtubule protein preparations under conditions of non-denaturing electrophoresis. Immunoprecipitation experiments with monoclonal antibodies against gamma-tubulin and alpha-tubulin revealed interactions of both gamma-tubulin forms with tubulin dimers, irrespective of the size of complexes. We suggest that, besides small and large gamma-tubulin complexes, other molecular gamma-tubulin form(s) exist in brain extracts. Two-dimensional electrophoresis revealed multiple charge variants of gamma-tubulin in both brain extracts and microtubule protein preparations. Post translational modification(s) of gamma-tubulins might therefore have an important role in the regulation of microtubule nucleation in neuronal cells. PMID- 11939911 TI - Superior prolonged antiemetic prophylaxis with a four-drug multimodal regimen - comparison with propofol or placebo. AB - BACKGROUND: The purpose of this study was to compare the effects of a low-dose propofol infusion with a four-drug multimodal regimen for prophylaxis of postoperative nausea and vomiting (PONV). METHODS: : PONV was studied in two patient groups with a known high incidence. Through a stratified randomization, 60 patients undergoing breast surgery and 120 patients undergoing abdominal surgery were randomized to three groups of equal size: the propofol group (P), the multidrug group (M) and the control group (C). All patients received general anesthesia, induction with propofol and maintenance with sevoflurane. After induction, patients in the P group received a continuous infusion of propofol 1 mg/kg/h during the operation and the first 4 postoperative h. Patients in the M group received dexamethasone 4 mg and three antiemetics, ondansetron 4 mg, droperidol 1.25 mg and metoclopramide 10 mg i.v. In the control group no prophylaxis was given. Nausea and pain were evaluated by incidence and a visual analog scale (0-10 cm). All emetic episodes were noted by the staff during the first 4 h and by the patients during the next 20 h. RESULTS: The overall incidence of PONV during the first 24 h postoperatively was significantly lower in the M group (24%) than in the P group (49%) (P<0.01) or the C group (70%) (P<0.001). The incidence of PONV increased significantly both in patients undergoing breast surgery and abdominal surgery after termination of propofol. The number of patients who vomited was significantly lower in the M group, both in breast surgery patients (5%) and abdominal surgery patients (3%) compared to patients in the propofol groups (breast 16% NS; abdominal 29%, P<0.05) and in the control groups (breast 37%, P<0.01; abdominal 29%, P<0.01). CONCLUSION: The incidence of PONV is very high in patients undergoing breast and abdominal surgery. In the present study antiemetic prophylaxis with a combination of droperidol, ondansetron, metoclopramide and dexamethasone was more effective in preventing PONV, especially vomiting, than a postoperative low-dose infusion of propofol, which had a short lasting effect. PMID- 11939913 TI - Comparison of auditory evoked potentials and the A-line ARX Index for monitoring the hypnotic level during sevoflurane and propofol induction. AB - BACKGROUND: Extraction of the middle latency auditory evoked potentials (AEP) by an auto regressive model with exogenous input (ARX) enables extraction of the AEP within 1.7 s. In this way, the depth of hypnosis can be monitored at almost real time. However, the identification and the interpretation of the appropriate signals of the AEP could be difficult to perform during the anesthesia procedure. This problem was addressed by defining an index which reflected the peak amplitudes and latencies of the AEP, developed to improve the clinical interpretation of the AEP. This index was defined as the A-line Arx Index (AAI). METHODS: The AEP and AAI were compared with the Modified Observers Assessment of Alertness and Sedation Scale (MOAAS) in 24 patients scheduled for cardiac surgery, anesthetized with propofol or sevoflurane. RESULTS: When comparing the AEP peak latencies and amplitudes and the AAI, measured at MOAAS level 5 and level 1, significant differences were achieved. (mean(SD) Nb latency: MOAAS 5 51.1 (7.3) ms vs. MOAAS 1: 68.6 (8.1) ms; AAI: MOAAS 5 74.9 (13.3) vs. MOAAS 1 20.7 (4.7)). Among the recorded parameters, the AAI was the best predictor of the awake/anesthetized states. CONCLUSION: We conclude that both the AAI values and the AEP peak latencies and amplitudes correlated well with the MOAAS levels 5 (awake) and 1 (anesthetized). PMID- 11939912 TI - Dimenhydrinate for prophylaxis of postoperative nausea and vomiting: a meta analysis of randomized controlled trials. AB - BACKGROUND: Diphenhydramine and its theoclate salt dimenhydrinate are traditional antiemetics still in use. However, so far the quantitative effect of dimenhydrinate in the prophylaxis of postoperative nausea and vomiting (PONV) has not been evaluated systematically. METHODS: Results from randomized controlled trials investigating the efficacy of dimenhydrinate vs. a control to prevent PONV were included in a meta-analysis. Studies were systematically searched through MEDLINE, EMBASE, the Cochrane-Library, manually screening of reference lists of matching review articles and current issues of locally available peer-reviewed anesthesia journals, up to June 2001. The numbers of patients with complete absence of PONV within 6 h and within 48 h after surgery were extracted as the main end point. Pooled relative benefits (RB) and numbers-needed-to-treat (NNT) with their corresponding 95% confidence intervals (CI) were calculated using a random effects model. This quantitative systematic review was performed following the recommendations of the QUORUM statement. In all, 18 trials with 3045 patients were included in the analysis: 1658 patients received a placebo (control) and 1387 patients received dimenhydrinate. RESULTS: The RB to stay completely free of PONV was 1.2 (95% CI: 1.1-1.4) for the early period (NNT = 8; 95% CI: 5-25) and 1.5 (1.3-1.8) for the overall investigated period (NNT = 5; 95% CI: 3-9). CONCLUSION: Dimenhydrinate is a traditional and inexpensive antiemetic with an efficacy that might be considered as clinically relevant. Although in use for a long time, the dose-response, precise estimation of side-effects, optimal time of administration and the benefit of repetitive doses still remain unclear. PMID- 11939914 TI - Effect of sevoflurane on the mid-latency auditory evoked potentials measured by a new fast extracting monitor. AB - BACKGROUND: Mid-latency auditory evoked potentials (MLAEP) are widely suppressed during general anesthesia and may therefore be useful for assessment of the depth of anesthesia. However, interpretation of amplitudes and latencies in the AEP signal is time consuming. A new monitor (A-line) that quantifies the MLAEP into an index has therefore been developed. The present study aimed to assess the precision of a prototype of the new monitor and to test the hypothesis that the depth of anesthesia index shows a graded response with changing steady-state end expiratory concentrations of sevoflurane. METHODS: We studied 10 ASA physical status I or II patients undergoing elective hysterectomy under combined epidural and general anesthesia by sevoflurane. Baseline auditory evoked potentials were recorded in the conscious patient immediately before induction of general anesthesia. Depth of anesthesia indices were recorded before anesthesia and at decreasing end-expiratory steady-state sevoflurane concentrations of 2.0%, 1.5%, 1.0% and 0.5%. All indices were recorded in duplicate 6 s apart. By use of an autoregressive model with exogenous input (ARX-model), the monitor extracted the AEP within 6 s. The depth of anesthesia AEP index calculated in this way was defined as the A-line ARX index (AAI). RESULTS: Approximately 95% of the differences between repeated recordings were 5 AAI-units or less. A wide interindividual variation was observed at each observation point. AAI at 1%, 1.5% and 2% end-expiratory concentration was significantly less than the baseline AAI obtained before induction of anesthesia (P < 0.001). AAI did not change significantly in the 1-2% concentration range. CONCLUSION: The new monitor was precise. Attenuation of the A-line ARX-index (AAI) for mid-latency auditory evoked potentials (MLAEP) during general anesthesia was profound. However, the monitor did not show a graded response with changing end-expiratory steady-state concentrations of sevoflurane. PMID- 11939915 TI - Two propofol formulations are equivalent in small children aged 1 month to 3 years. AB - BACKGROUND: Propofol has been widely used in general anesthesia. Although it is also often used in pediatric anesthesia, there has been only limited scientific evidence on the use of propofol in children up to 3 years. METHODS: A prospective, randomized, double-blind, therapeutic equivalence study comparing two propofol 1% emulsions (Propofol 1% Fresenius vs. Diprivan 1%) was performed in 60 patients scheduled for routine surgery or for diagnostic laparoscopic procedures requiring anesthesia. To guarantee comparability of age distribution between the two groups, a stratified randomization with patients younger than 12 months of age in a low age group and with patients aged 12 months to 3 years in a high age group was used. The average propofol induction dose and the average propofol infusion dose were analyzed to prove equivalence. The side-effects profile was analyzed to compare the safety profiles of the two propofol formulations in this study. RESULTS: There were no differences in baseline characteristics between the two treatment groups of high and low age. Medications used for induction and maintenance of anesthesia, and side-effects profiles were comparable, as were the average propofol dose for induction of anesthesia (range of the mean dose 4.0-4.2 mg/kg) and for maintenance of anesthesia (range of the mean dose in the first hour 8.74-9.42 mg x kg(-1) x h(-1)). CONCLUSIONS: The two 1% propofol formulations were equally effective in our patient population of infants and young children between 1 month and 3 years of age. No differences between the two propofol formulations were found with regard to the circulatory reaction, lipid metabolism, dosages, and recovery profile in the studied age groups. PMID- 11939916 TI - Treatment of intracranial hypertension and aspects on lumbar dural puncture in severe bacterial meningitis. AB - BACKGROUND: Brain stem herniation due to raised intracranial pressure (ICP) is a common cause of mortality in severe bacterial meningitis, but continuous measurements of ICP and the effects of ICP-reducing therapy in these patients have, to our knowledge, not been described. METHODS: During a four-year period, an ICP-monitoring device was implanted in patients admitted to our hospital with severe bacterial meningitis and suspected intracranial hypertension. ICP above 20 mmHg was treated using the Lund Concept, which includes antihypertensive therapy (beta1-antagonist,alpha2-agonist), normalization of the plasma colloid osmotic pressure and the blood volume, and antistress therapy. RESULTS: ICP above 20 mmHg was found in all 12 patients studied. It was effectively reduced in all but two patients, who died. Both patients had a low cerebral perfusion pressure (<10 mmHg), dilated pupils at start of therapy and were beyond recovery. Radiological signs of brain swelling were present in only five patients. Seven patients recovered fully, while mild audiological impairment was observed in two and minor neurological sequelae in one patient. Eight patients showed signs suggesting imminent brain stem herniation before start of ICP-reducing treatment, seven of whom had been subjected to diagnostic lumbar dural puncture shortly before development of the brain stem symptoms. These symptoms gradually regressed after initiation of therapy, and in one patient reversal of brain stem herniation was documented by MRI. CONCLUSIONS: Severe bacterial meningitis can be associated with increased ICP, which can be reduced using the Lund Concept. The high survival rate, the low frequency of sequelae and the reversal of signs of imminent brain stem herniation in these high-risk patients indicated beneficial effects of the intervention. The study confirms earlier observations that lumbar dural puncture is potentially hazardous in patients with intracranial hypertension, because it may trigger brain stem herniation. A normal CT brain scan does not rule out intracranial hypertension. PMID- 11939917 TI - Myocardial ischemia induces coronary t-PA release in the pig. AB - BACKGROUND: Tissue-type plasminogen activator (t-PA) is the key factor in initiating endogenous fibrinolysis in the vascular compartment. Regulated release of t-PA from endothelial stores is rapidly induced by several humoral factors as well as coagulation activation products. The aim of the present study was to test the hypothesis that regional myocardial ischemia induces regulated release of t PA in the coronary vasculature in vivo. METHODS: Healthy anesthetized (pentobarbital) pigs (n=8) were studied before and after a 10-min left anterior descending region coronary artery occlusion (LAD). Coronary fluxes of lactate, total t-PA antigen (ELISA, detecting both complex bound and free fraction) and active t-PA (functional assay detecting biological free fraction) were determined at 1, 3, 5 and 10 min of reflow. RESULTS: Coronary occlusion induced myocardial lactate production in all animals. Net coronary release of total t-PA, which was 21 ng/min during control, increased rapidly during reflow with a peak after only 1 min (136 ng/min), and returned to baseline within 3 min. Net release of active t-PA mirrored the overall net release response, but fell short of statistical significance. CONCLUSION: Data indicate a local myocardial profibrinolytic response following regional ischemia, which may serve as a prompt defence against coronary thromboembolic events. PMID- 11939918 TI - Detecting postoperative urinary retention with an ultrasound scanner. AB - BACKGROUND: Retention of urine is a common postoperative problem associated with risk of overdistention and permanent detrusor damage. Prevention of urinary retention by insertion of indwelling catheter may increase the risk of urinary infection. We have performed a reliability test of an ultrasound scanner, implemented in the postoperative monitoring equipment. METHODS: Patients were monitored after different types of surgery under spinal anesthesia with an ultrasound scanner in the postanesthesia care unit (PACU). PATIENTS: Patients who according to current guidelines required a urinary bladder catheter, were scanned before a catheter was inserted and urine volume was measured. These two urine volumes were compared and analyzed for agreement. RESULTS: Nineteen female and 17 male patients were included. The mean difference between ultrasound estimates and catheter urine volume measurements was - 21.5 mL, and limits of agreement, calculated as a 95% confidence interval, were - 147 and + 104 mL. This means that the urine volume estimated by ultrasound was on average 21.5 mL smaller than the urine volume when the bladder was emptied. CONCLUSION: This study confirms a good agreement between the ultrasound scanner estimates of urinary bladder volume and urine volume measured after emptying the bladder. Nurses in the PACU could operate the ultrasound scanner after a brief instruction and training period. Considering the potentially serious long-term consequences of undiagnosed postoperative urinary retention, introducing this equipment for routine monitoring of urinary bladder volume should be considered. PMID- 11939919 TI - Effect of rewarming speed during hypothermic cardiopulmonary bypass on cerebral pressure-flow relation. AB - BACKGROUND: Cerebral blood flow is less dependent on arterial blood pressure during hypothermic cardiopulmonary bypass (CPB) compared to warm CPB. Fast rewarming has a more pronounced effect on cognitive performance in the elderly and causes an increased arterio-jugular oxygen content difference. We studied the effect of rewarming and rewarming speed on cerebral pressure-flow relation in adult patients undergoing elective coronary artery bypass surgery with mild hypothermic CPB. METHODS: Fifty patients were randomly assigned to either a slow rewarming strategy (0.24 degrees C/min) or a fast rewarming strategy (0.5 degrees C/min). Cerebral pressure-flow relation was assessed by a transcranial Doppler derived index for cerebral pressure-flow relation (Pressure-flow Index, PFI). The effect of rewarming speed on cerebral pressure-flow relation was assessed by comparing the absolute PFI value after rewarming between the two treatment groups. RESULTS: The mean PFI decreased significantly from 0.73 (standard deviation: 0.28) before rewarming to 0.54 (0.35) after rewarming in the slow rewarming group and from 0.63 (0.29) to 0.48 (0.30) in the fast rewarming group. Absolute PFI after rewarming was not significantly different (mean PFI difference = 0.06; 95% CI = - 0.13; 0.26) between both rewarming strategies. CONCLUSION: Rewarming from mild hypothermic CPB might result in pressure-dependent cerebral blood flow velocity but rewarming speed did not aggravate the effect of rewarming on pressure-flow dependency. PMID- 11939920 TI - Computer-aided ventilator resetting is feasible on the basis of a physiological profile. AB - BACKGROUND: Ventilator resetting is frequently needed to adjust tidal volume, pressure and gas exchange. The system comprising lungs and ventilator is so complex that a trial and error strategy is often applied. Comprehensive characterization of lung physiology is feasible by monitoring. The hypothesis that the effect of ventilator resetting could be predicted by computer simulation based on a physiological profile was tested in healthy pigs. METHODS: Flow, pressure and CO2 signals were recorded in 7 ventilated pigs. Elastic recoil pressure was measured at postinspiratory and post-expiratory pauses. Inspiratory and expiratory resistance as a function of volume and compliance were calculated. CO2 elimination per breath was expressed as a function of tidal volume. Calculating pressure and flow moment by moment simulated the effect of ventilator action, when respiratory rate was varied between 10 and 30 min(-1) and minute volume was changed so as to maintain PaCO2. Predicted values of peak airway pressure, plateau pressure, and CO2 elimination were compared to values measured after resetting. RESULTS: With 95% confidence, predicted pressures and CO2 elimination deviated from measured values with < 1 cm H2O and < 6%, respectively. CONCLUSION: It is feasible to predict effects of ventilator resetting on the basis of a physiological profile at least in health. PMID- 11939921 TI - Spasmolytic effects of colforsin daropate on serotonin-induced pulmonary hypertension and bronchoconstriction in dogs. AB - BACKGROUND: We have previously found that agents increasing intracellular cAMP levels of smooth muscles, such as PDE3 inhibitors, aminophylline and prostaglandin E1, produce both bronchodilation and pulmonary vasodilation in serotonin-induced pulmonary hypertension and bronchoconstriction models. In the present study we have simultaneously evaluated the spasmolytic effects of colforsin daropate, a novel forskolin derivative, on serotonin-induced pulmonary hypertension and bronchoconstriction. METHODS: Ten mongrel dogs were anesthetized with pentobarbital. The pulmonary hypertension and bronchoconstriction were elicited with serotonin (10 microg/kg + 1 mg x kg(-1) x h(-1)) and assessed as percentage changes in pulmonary vascular resistance (PVR) and bronchial cross sectional area (BCA) (basal = 100%). Initially, the relaxant effects of colforsin daropate (0-300 microg/kg) were determined. The PVR and BCA were assessed before and 30 min after serotonin infusion began and 5 min after each dose of colforsin daropate. To determine whether colforsin daropate-induced relaxation is independent of plasma catecholamine, propranolol 0.4 mg/kg was given following colforsin daropate 300 microg/kg i.v. RESULTS: Colforsin daropate reversed both pulmonary hypertension and bronchoconstriction dose-dependently: -logED50 (95% confidence intervals, mean ED50) for pulmonary hypertension and bronchoconstriction 5.44 (5.08-5.80, 3.6 microg/kg) and 4.90 (4.06-5.20, 12.5 microg/kg), respectively. However, colforsin daropate (>or= 30 microg/kg) produced a more pronounced systemic than pulmonary vasodilation. Although colforsin daropate (>or= 30 microg/kg) significantly increased plasma catecholamines, propranolol did not reverse the relaxant effects. CONCLUSIONS: Colforsin daropate may attenuate bronchoconstriction and pulmonary hypertension. In addition, as beta-blockade did not change the attenuation, the relaxant effects may be independent of plasma catecholamines. PMID- 11939922 TI - Changes in intrathoracic blood volume associated with pneumoperitoneum and positioning. AB - BACKGROUND: It is still controversial whether elevated cardiac filling pressures after the onset of pneumoperitoneum are the consequence of increased intrathoracic pressure or of increased venous return. The aim of this study was to assess the effects of pneumoperitoneum and body positioning on intrathoracic blood volume (ITBV). METHODS: Thirty anesthetized patients were randomly assigned to have CO2-pneumoperitoneum (13 mmHg) either in a supine, in a 15 degrees head up tilt or in a 15 degrees head-down tilt position. Measurements of ITBV and hemodynamics by the double indicator method were recorded after induction of anesthesia and application of a fluid bolus (Lactated Ringer's solution 10 ml/kg), after positioning and after induction of pneumoperitoneum. RESULTS: Intrathoracic blood volume index (ITBVI) increased significantly after induction of pneumoperitoneum in all body positions (supine: from 18.5 +/- 3.3 -20.2 +/- 5.2 ml/kg (+6%) head-up from 16.7 +/- 3.8 - 17.4 +/- 3.7 ml/kg (+16%) and head down: from 19.8 +/- 5.6 - 20.5 +/- 5.9 ml/kg (+14%)). Heart rate did not change significantly in any of the groups. Cardiac index showed a statistically significant change in the head-down position with pneumoperitoneum (-11%). A good correlation was found for stroke volume (SV) with ITBV (r = 0.79), but not with central venous pressure (r = 0.26). Systemic vascular resistance index increased significantly in all three groups (supine +6%, head-up +16%, head-down position +14%). CONCLUSION: The present study indicates that the onset of pneumoperitoneum, even with moderate intra-abdominal pressures, is associated with an increased intrathoracic blood volume in ASA I/II patients. PMID- 11939923 TI - Remifentanil sedation compared with propofol during regional anaesthesia. AB - BACKGROUND: The short onset and offset of remifentanil may allow for accurate dosing of sedative effect with few side-effects and rapid recovery. In this study remifentanil is compared with propofol for sedation during successful regional anaesthetic blocks. METHODS: After informed consent was given, 125 patients undergoing surgery under spinal or brachial plexus anaesthesia were randomized to receive, either propofol: bolus 500 microg/kg plus initial infusion 50 microgkg/min or remifentanil: bolus 0.5 microg/kg plus initial infusion 0.1 microgkg/min. Study drug infusion rate was titrated throughout the procedure according to level of sedation and side-effects. Pain, discomfort, sedation level and side-effects were recorded at regular intervals until discharge from the post operative care unit (PACU). RESULTS: Two patients in the remifentanil group versus ten in the propofol group were treated for discomfort or pain during surgery (P<0.02). Due to a significantly higher rate of respiratory depression (46% vs. 19% with propofol, P<0.01) the mean remifentanil infusion rate was decreased to 0.078 +/- 0.028 microgkg/min, whereas it was kept stable with propofol. Propofol patients had significantly higher (P<0.05) sedation levels and experienced more frequent amnesia of the procedure. Eleven propofol patients experienced pain at injection site, versus two remifentanil patients (P<0.02). Nausea and vomiting were more frequent in the remifentanil patients during infusion (27% vs. 2% in the propofol group, P<0.001) but similar postoperatively. Time to discharge from PACU was similar in the two groups. CONCLUSION: Propofol results in less respiratory depression and nausea when sedation is needed during a case with a successful regional block. Remifentanil may be considered as an alternative if pain during the procedure is a major concern or if amnesia is contraindicated. PMID- 11939924 TI - Ropivacaine 2 mg/mL vs. bupivacaine 1.25 mg/mL with sufentanil using patient controlled epidural analgesia in labour. AB - BACKGROUND: In recent studies, minimum local analgesic concentrations have been defined as 0.93 mg/mL for bupivacaine and 1.56 mg/mL for ropivacaine for epidural analgesia for the first stage of labour, resulting in an analgesic potency ratio of 1 : 0.6. In the current study we compared ropivacaine and bupivacaine in a PCEA system (combined with sufentanil) taking this potency ratio into account but administering drug doses providing sufficient analgesia for all stages of labour. METHODS: In a prospective, double-blinded study 114 parturients were randomised to receive either ropivacaine 2 mg/mL with sufentanil 0.75 microg/mL or bupivacaine 1.25 mg/with sufentanil 0.75 microg/mL. After epidural catheter placement, PCEA was available with boluses of 4 mL, a lock-out time of 20 min and no basal infusion rate. We evaluated pain intensity during contractions, sensory and motor function, duration of labour, mode of delivery and neonatal outcome. Consumption of local anaesthetic and opioid drugs and PCEA system variables were recorded. RESULTS: Mean total consumption as well as mean hourly drug consumption was significantly increased in the ropivacaine-sufentanil group. No differences in analgesic quality, sensory or motor blocking potencies or neonatal outcome variables between groups were detected. Frequency of instrumental deliveries was significantly increased in the ropivacaine-sufentanil group. CONCLUSIONS: The results support the findings of previously published studies postulating ropivacaine to be 40-50% less potent for labour epidural analgesia compared to bupivacaine. However, we observed an increased frequency of instrumental deliveries with ropivacaine. To evaluate the clinical relevance of these findings, further investigations are warranted. PMID- 11939925 TI - Comparison of the opioid-sparing efficacy of diclofenac and ketoprofen for 3 days after knee arthroplasty. AB - BACKGROUND: Comparative postoperative non-steroidal anti-inflammatory drug (NSAID) studies in orthopedic patients have usually been restricted in time to the first postoperative day. The opioid-sparing effect of NSAIDs may be beneficial postoperatively as long as pain otherwise restricts ambulation and rehabilitation. We therefore compared the analgesic efficacy of the maximum recommended doses of diclofenac and ketoprofen for 3 days after knee arthroplasty. METHODS: We studied 64 knee arthroplasty patients, operated on under spinal anesthesia. In a randomized, double-blind and placebo-controlled fashion, the patients received either i.v. diclofenac 75 mg (n = 24), ketoprofen 100 mg (n = 24) or saline (n = 16) in the recovery room, followed by oral diclofenac 150 mg/day, ketoprofen 300 mg/day or placebo, respectively, for 3 days, supplemented by patient-controlled analgesia (PCA) with i.v. oxycodone. RESULTS: The mean consumption of oxycodone during the first, second and third study days was 45.3, 22.3 and 15.2 mg in the diclofenac group, 43.5, 37.5 and 21.8 mg in the ketoprofen group, and 61.2, 45.9 and 36.1 mg, respectively, in the placebo group. Oxycodone consumption was significantly lower (P < 0.05) in the ketoprofen group than in the placebo group in the postoperative period 13-24 h and 61-72 h. Diclofenac was superior to placebo in the postoperative period 25-48 h (P < 0.01), 49-60 h (P < 0.05) and to ketoprofen at 49-60 h (P < 0.05). During administration of diclofenac on days 1-3 and ketoprofen on day 2, the mean pain scores (VAS) were lower than in the placebo group (P < 0.05). Six patients had difficulties in operating the PCA device. There were no differences in blood loss. CONCLUSION: We conclude that in the first day after knee arthroplasty (13 24 h), ketoprofen exerted an opioid-sparing effect. After day 1 (25-60 h), with the doses used, diclofenac proved to be better than placebo, whereas ketoprofen was not. PMID- 11939926 TI - Continuous spinal analgesia for labor and delivery in a parturient with hypertrophic obstructive cardiomyopathy. AB - Induction of labor under analgesia was planned for a 30-year-old-primiparous patient with hypertrophic obstructive cardiomyopathy (HOCM), as her fetal evaluation revealed intrauterine growth restriction at 38 weeks' gestation. However, regional analgesia during labor may present a potential risk for hemodynamic instability in patients with HOCM due to the possibility of a sympathetic block, as a result of vasodilation associated with the administration of local anesthesia. This case report demonstrates the successful management of the patient with analgesia provided by a continuous spinal catheter dosed with a continuous infusion of fentanyl and supplemental meperidine. Fetal surveillance monitoring included fetal pulse oximetry in addition to conventional cardiotocography, on the basis of which cesarean section was avoided. PMID- 11939927 TI - One-lung ventilation of a preterm newborn during esophageal atresia and tracheoesophageal fistula repair. AB - In this paper, we assessed the anesthesia management of a male, a 34-week gestation age newborn, weighing 1500 g, who has esophageal atresia and tracheoesophageal fistula localized just above the carina. Endotracheal intubation and intermittent positive pressure ventilation caused air leakage through the fistula into the stomach, causing abdominal distention. One-lung ventilation by left main bronchus intubation eliminated this problem. PMID- 11939928 TI - Fatal amniotic fluid embolism diagnosed histologically. AB - Although the mortality of amniotic fluid embolism remains very high, there are more and more reports of patients surviving after prompt and aggressive therapy. However its pathogenesis is controversial, and, in some cases, an anaphylactic reaction to the amniotic fluid rather than the hemodynamic changes following mechanical obstruction by an embolus has been proposed as the underlying pathophysiological mechanism. In this paper we describe a case of amniotic fluid embolism with florid clinical features and a difficult and delayed diagnosis. On autopsy, clear-cut histological changes of amniotic fluid embolism were found. PMID- 11939929 TI - Closed suctioning systems. PMID- 11939930 TI - Thoracic epidural analgesia increases vital capacity after cardiac surgery. PMID- 11939931 TI - Combitude and difficult intubation. PMID- 11939933 TI - 'Tricks of the trade' with the Esophageal-Tracheal Combitube. PMID- 11939934 TI - New insertion technique of the Esophageal-Tracheal Combitube. PMID- 11939935 TI - Learning capnography on the World Wide Web: an educational resource for the new millennium. PMID- 11939936 TI - Efficacy and safety of entacapone in Parkinson's disease patients with suboptimal levodopa response: a 6-month randomized placebo-controlled double-blind study in Germany and Austria (Celomen study). AB - OBJECTIVES: To determine the efficacy and safety of the catechol-O methyltransferase (COMT) inhibitor entacapone, used as an adjunct to levodopa, in Parkinson's disease (PD) patients. PATIENTS AND METHODS: In this parallel group, randomized, double-blind study, 301 PD patients, the majority with motor fluctuations, received entacapone (200 mg) or placebo with each daily dose of standard or controlled-release (CR) levodopa. The 24-week treatment period was followed by 2 weeks of entacapone withdrawal. Efficacy was determined by home diaries ('on' and 'off' times), Unified Parkinson's Disease Rating Scale (UPDRS) and changes in levodopa dosage, and safety by adverse-event inquiry, vital signs, electro cardiography (ECG) and laboratory tests. RESULTS: In the total population, the UPDRS activities of daily living and motor scores were significantly improved (P < 0.05) by entacapone vs placebo. In fluctuating patients, 'on' time increased (1.7 h) and 'off' time decreased (1.5 h) significantly more with entacapone than with placebo (0.5 and 0.6 h, respectively; P < 0.05), and the daily levodopa dose was reduced by 54 mg with entacapone and increased by 27 mg with placebo (P < 0.05). Entacapone benefit was lost on withdrawal. Entacapone efficacy was comparable between patients using CR and standard levodopa preparations. Increased dyskinesias (entacapone 34%, placebo 26%) and nausea (10 and 5%, respectively), mostly occurring shortly after treatment initiation, were generally managed by reducing the levodopa dose. Diarrhoea (entacapone 8%, placebo 4%) was seldom severe. There were no differences in vital signs, ECG or laboratory results. CONCLUSION: Entacapone is an effective and safe levodopa extender and enhancer, improving the symptomatic efficacy of levodopa in PD and adding to the patients' benefit. PMID- 11939937 TI - Progress of visual dysfunction in Parkinson's disease. AB - Studies on progression of Parkinson's disease (PD) mainly focus on the nigrostriatal dopaminergic decline, but not on the visual system. We determined progression of (i) disturbed color vision, assessed with the Farnsworth-Munsell 100 Hue test (FMT) and (ii) intensity of PD in 18 patients. Significant differences occurred between (i) initial FMT error scores and follow-up results 3 years later (P=0.002) and analogously (ii) scored intensity of PD (P=0.002). A relation between computed differences of FMT error scores and rated activities of daily living appeared. Deterioration of color vision progresses in PD. PMID- 11939938 TI - Diagnostic patterns of regional atrophy on MRI and regional cerebral blood flow change on SPECT in young onset patients with Alzheimer's disease, frontotemporal dementia and vascular dementia. AB - OBJECTIVES: Alzheimer's disease (AD), frontotemporal dementia (FTD) and vascular dementia (VaD) are the three most common causes of young onset dementias. Most neuroimaging studies of these disorders have involved comparisons with normal controls. The aims of this study were to examine the clinical diagnostic value of magnetic resonance imaging (MRI) and single photon emission computed tomography (SPECT) (in combination and in isolation) in the differentiation of one form of dementia from another from amongst a group of AD, FTD and VaD. METHODS: T1 weighted MRI images and 99mTc-HMPAO SPECT images were obtained from consecutive patients with FTD (n=21), AD (n=23) and VaD (n=20) and rated visually by experienced neuroradiologists and nuclear medicine physicians. RESULTS: Asymmetrical atrophy was seen only in FTD. Frontotemporal dementia patients were the most atrophic whereas severe atrophy was rarely observed in VaD. Severe frontal atrophy (unilaterally or bilaterally) and/or asymmetrical atrophy on MRI is highly diagnostic (sensitivity 0.71, specificity 0.93, LR 10.24) of FTD from within a group of FTD and non-FTD (AD, VaD) patients. Mild or severe parietal atrophy with severe reduction in parietal regional cerebral blood flow on SPECT is diagnostic (sensitivity 0.71, specificity 0.76, LR 3.02) of AD from within a group of AD and non-AD (VaD, FTD) patients. CONCLUSION: Anatomical (MRI) and functional (SPECT) imaging provide different information and a combination of these modalities improves diagnostic specificity. PMID- 11939939 TI - Chronic alcoholism and the frontal lobe: which executive functions are imparied? AB - OBJECTIVE: Over the last decade, various hypotheses have been advanced concerning the cognitive functions affected by chronic alcoholism. The aim of this study was to identify the pattern of executive function impairment in chronic alcoholism, shedding light on possible differences between specific functions related to the frontal lobe. METHODS: Twenty-two male alcoholics and 22 controls, matched for age, educational level and IQ, were enrolled in the study. MMPI and a battery of neuropsychological tests [i.e. digit symbol, trail making test, Stroop test, digit cancellation test, Wisconsin card sorting test (WCST), simple and choice reaction times] for assessing frontal lobe functioning were administered. RESULTS: The alcoholics were found to be impaired in a wide range of executive domains, with the exception of the Stroop test, which nevertheless showed a trend towards statistically significant differences between patients and controls. CONCLUSION: With the exception of aggression - our subjects did not have high aggression scale scores - the 'frontal lobe hypothesis', according to which alcoholic patients are impaired on function tests related to the frontal lobe, was therefore confirmed in our sample. PMID- 11939940 TI - Site of bleeding and early outcome in primary intracerebral hemorrhage. AB - OBJECTIVES: To describe the influence of the site of the bleeding on clinical spectrum and early outcome of patients with acute spontaneous intracerebral hemorrhage (ICH). MATERIALS AND METHODS: ICH was diagnosed in 229 (11%) of 2000 consecutive stroke patients included in a prospective stroke registry over a 10 year period. Frequency of demographic variables, risk factors, clinical events, neuroimaging data, and early outcome (until hospital discharge) according to different sites of bleeding was assessed. Each topography of the bleeding (independent variable) was compared with the remaining ICH cases by means of logistic regression analysis. RESULTS: In the multivariate analysis, sensory deficit was significantly associated with ICH in the thalamus; lacunar syndrome and hypertension with ICH in the internal capsule-basal ganglia; seizures, non sudden stroke onset, and hypertension with lobar ICH; ataxia and sensory deficit with ICH in the cerebellum; cranial nerve palsy with ICH in the brainstem; and limb weakness, diabetes, and altered consciousness with multiple topographic involvement. The overall in-hospital mortality rate was 31%, but this varied from 65% for multiple topographic involvement, 44% for intraventricular ICH, and 40% for ICH in the brainstem to 16% for ICH in the internal capsule-basal ganglia. CONCLUSION: These data show the heterogeneous clinical profile of ICH, but they also suggest a difference in the clinical spectrum and in-hospital mortality according to the site of bleeding. PMID- 11939941 TI - Surgical treatment of patients with low-grade astrocytomas and medically intractable seizures. AB - OBJECTIVE: Low-grade astrocytomas can present with seizures that respond poorly to antiseizure medications, with a consequent reduction in the quality of life, because of both seizures and the medication's side-effects. We report our experience with operative treatment of such patients. SUBJECTS AND METHODS: Five patients (two children and three adults) with supratentorial low-grade astrocytomas associated with severe seizures were operated on. We followed the effects of the operation on post-operative neurological deficit, seizure frequency and the quality of life. RESULTS: No serious neurological complications followed the operations. The patients were followed for a median period of 12 months (6-46). Post-operatively, three of the patients became seizure-free, one experiences only auras, and one had a great reduction in seizure frequency and severity. All patients reported great improvement in their quality of life. CONCLUSION: Resections of low-grade astrocytomas in patients with medically intractable seizures are safe procedures that effectively control seizures in the majority of patients, resulting in significant improvement in the patients' quality of life. PMID- 11939942 TI - Parental responses to first and recurrent febrile convulsions. AB - OBJECTIVE: To compare parental knowledge, attitudes, concerns, and first-aid practices for children experiencing febrile convulsions (FC). SUBJECTS AND METHODS: A questionnaire was mailed to 326 FC parents from 11 emergency departments in southern Taiwan. RESULTS: A total of 109 first- and 107 recurrent FC parents responded 1-3 months after the FC. Most incorrectly believed an electroencephalogram (EEG) or computed tomography (CT) was necessary; immunization should be postponed, and overestimated the risk of subsequent epilepsy. Parents were concerned about further attacks in the night, fever episodes, and frequently measured the child's body temperature. During the first episode, objects were inserted into the child's mouth and they were rushed to a hospital. One third lowered the child's body temperature, and 15% positioned the children on their side. For subsequent seizures, 80% anticipated rushing the child to a hospital, and 44% would put objects into the child's mouth. In comparison, although the recurrent-FC parents had higher scores in knowledge and attitudes than the first-FC parents, low knowledge scores (40% correct) were seen in both groups. No significant differences were found on parental concerns, performed/anticipated first aid for FC. CONCLUSION: Most FC parents had inadequate knowledge, high concerns, and improper first-aid practices. This suggests that parents with either first- or recurrent-FC children need information, emotional support, and first-aid demonstrations. PMID- 11939943 TI - Serum cytokine levels do not correlate with disease activity and severity assessed by brain MRI in multiple sclerosis. AB - OBJECTIVE: Chronic and acute dysregulation of the cytokine network has been described in multiple sclerosis (MS). Inflammatory lesions in the central nervous system of MS patients can be assessed by brain magnetic resonance imaging (MRI). This study has been performed to investigate whether changes of cytokines correlate with morphological changes as determined by MRI. MATERIALS AND METHODS: We included 46 patients with relapsing-remitting MS in the study. The serum concentrations of tumor necrosis factor-beta (TNF-beta), TNF receptor-1 (TNFR-1; 55 kDa) and TNFR-2 (75 kDa), interleukin-4 (IL-4), interleukin-10 (IL-10) and interferon-gamma (IFN-gamma) were measured by enzyme linked immunosorbent assay in all patients. Each parameter was correlated with clinical findings and brain MRI parameters. We measured both the number (lesion load) and cumulated area (disease burden) of all lesions on brain MRI. In addition, the number and cumulated area of those lesions showing signs of activity [Gadolinium (Gd) enhancement, perifocal edema] were determined. RESULTS: A non-significant trend (P < 0.05) was found only for the correlation of serum IFN-gamma levels and the number of active MRI lesions showing both Gd-enhancement and perifocal edema in the subgroup of patients (n=21) with active lesions. When corrected for multiple comparisons, this correlation was not significant anymore, as it was above the corrected P-value of 0.001. We could not observe any further correlation of cytokine levels and MRI parameters. However, TNF-beta serum levels were significantly (P < 0.05) elevated in the patient subgroups with higher number of lesions and disease burden, respectively. CONCLUSION: Our data show that the determination of serum levels of the investigated cytokines and cytokine receptors is not useful as a tool to determine subclinical disease activity and severity as assessed by brain MRI. PMID- 11939944 TI - Matrix metalloproteinases in inflammatory myopathies: enhanced immunoreactivity near atrophic myofibers. AB - OBJECTIVES: To further examine the role of proteolytic enzyme expression of matrix metalloproteinases (MMP) and T-cell markers in inflammatory myopathies and controls. MATERIAL AND METHODS: We studied the expression of MMP-2, MMP-7, and MMP-9 in 19 cases of inflammatory myopathies and controls using immunocytochemistry. RESULTS: Inflammatory myopathies showed distinct patterns of up-regulation of MMP. MMP-9 was strongly expressed in atrophic myofibers in all inflammatory myopathies. MMP-2 immunoreactivity was similar in its distribution, however, to a weaker intensity. In dermatomyositis the perifascicular atrophy showed pronounced MMP-9 immunoreactivity, probably reflecting denervated patterns of myofibers. Moreover, MMP-7 strongly immunolabeled invaded myofibers in polymyositis cases only. CONCLUSION: These patterns confirm, that MMP-7 up regulation is prominent in PM, while MMP-2 immunoreactivity is only slightly elevated in inflamed muscle. In general, MMP-9 up-regulation appears to be an important additional molecular event in the multistep process of all inflammatory myopathies. PMID- 11939945 TI - Fibrillation potentials in paraspinal muscles in chronic lumbosacral radiculopathy. AB - To assess the rate of fibrillation and/or positive sharp waves (FPSW) in the paraspinal muscles (PM) of patients with chronic lumbosacral radiculopathy, PM and leg muscles of 179 patients, 111 men and 68 women, aged 24-68 years (mean 49.0 +/- 9.3), were tested. Illness duration ranged from 8 months to 40 years (9.4 +/- 7.5 years). FPSW were detected in 38 patients (21.2%). In 28 of them (15.6%), FPSW were present in the limb muscles only, in eight (4.5%) in both PM and limb muscles, and in two (1.1%) solely in the PM. FPSW were accompanied in all the above patients by chronic neurogenic changes of motor unit potentials. Logistic regression analysis did not show any significant difference between: (i) the patients with and without fibrillations in leg and/or PM muscles, and (ii) between those with FPSW in both the PM and leg muscles and those with FPSW in the leg muscles only, when the effect of age and duration of illness were considered. These suggest that the paucity of FPSW in the PM is rather genuine and does not depend on the timing of the examination. In consequence, PM electromyography seems to be not as valuable in chronic radiculopathy as it is in the acute stage. PMID- 11939946 TI - A case of Kleine-Levin syndrome examined with SPECT and neuropsychological testing. AB - A case of Kleine-Levin syndrome with typical periodic hypersomnia and bulimia was diagnosed. On examination with single photo emission tomography (SPECT) (CERETEC) during a relapse period and 2 weeks later there was marked cortical hypoperfusion of the frontal and temporal lobes, especially on the left side as well as in the right parietal lobe. Neuropsychological testing performed 1 week after a relapse showed a reduction in encoding to memory function of verbal learning indicating neocortical damage of the left fronto-temporal region. A follow-up 2 months later after the patient had spontaneously recovered showed only a slight left fronto temporal disturbance. CT and MRI of the brain were normal although the MRI showed a large and asymmetric mamillary body. Neuropsychological testing 6 years after recovery showed pronounced reduction in short-time verbal and visual memory. Seven years after recovery SPECT demonstrated a normalized frontal perfusion but still a slight hypoperfusion in the left temporal lobe. Our results correlate to autopsy findings in two cases described previously. PMID- 11939947 TI - A case of idiopathic spinal epidural lipomatosis presented with radicular pain caused by compression with enlarged veins surrounding nerve roots. AB - OBJECTIVE: To report a case of idiopathic spinal epidural lipomatosis (SEDL) presented with unique radicular pain most likely caused by enlarged veins surrounding nerve roots. PATIENT: A 26-year-old male presented with radicular pain of the right T6-T7 area. He also showed Becker's nevus in the corresponding area. CT myelography and magnetic resonance imaging revealed epidural lipomatosis posterior to T4-T8 of the spinal cord. Surgical removal of adipose tissue and a hemilaminectomy of T4-T7 were performed and resulted in relief of the radicular pain. CONCLUSIONS: Lipomatosis was histologically confirmed and surrounded by enlarged veins. These abnormally enlarged veins compressed the nerve roots and were thought to cause radicular pain. Also, Becker's nevus of this case seems to have some relationship with SEDL. PMID- 11939948 TI - Spinal accessory mononeuropathy following posterior fossa decompression surgery. AB - Isolated injury of the spinal accessory nerve is a well-recognized complication of surgeries involving the posterior triangle of the neck. The procedures most commonly implicated are lymph node biopsy and carotid endarterectomy. We present a patient with isolated injury to the spinal accessory nerve, localized proximal to the innervation of the sternocleidomastoid muscle, which was noted following suboccipital decompression for an Arnold-Chiari malformation. To our knowledge, this association has not been previously reported. PMID- 11939949 TI - Dissection of the intracranial internal carotid artery producing isolated oculomotor nerve palsy with sparing of pupil. AB - Dissection of the internal carotid artery usually occurs in the cervical segment, but rarely may involve the artery in the intracranial course (1). The clinical course of intracranial dissection is often catastrophic, with rapid onset of profound neurological deficit, as a result of middle and/or anterior cerebral artery involvement. When this occurs the mortality rate is generally considered high. We describe a case of intracranial internal carotid artery dissection following trivial trauma presented with an isolated painful pupillary sparing oculomotor nerve palsy. PMID- 11939950 TI - Acute disseminated encephalomyelitis following nephropathia epidemica. AB - Acute disseminated encephalomyelitis (ADEM) is an acute monophasic inflammatory and demyelinating disease of the central nervous system (CNS) occurring days to weeks after a virus infection or vaccination. Nephropathia epidemica (NE) is a haemorrhagic fever with renal syndrome caused by Puumala virus, with endemic regions in Europe, especially Scandinavia and Western Russia. We describe a case of severe nephropathia epidemica requiring dialysis, followed by severe CNS symptoms caused by ADEM. To our best knowledge this is the first case in the literature in which NE caused ADEM. PMID- 11939951 TI - Cruciate paralysis or man-in-the-barrel syndrome? Report of a case of brachial diplegia. AB - A patient who developed isolated brachial diplegia following cardiac surgery is described. The underlying cerebral lesion could not be localized using magnetic resonance imaging (MRI). Evoked potentials disclosed normal findings, while pathological latencies were seen on cortical magnetic stimulation. Their marked improvement over the following year was accompanied by almost complete clinical recovery. The preserved arm reflexes, together with the observed slow firing motor units in electromyography argued against bilateral lesions of the brachial plexus. We attribute the observed diplegia to a medullary lesion at the level of the pyramidal decussation, presumably caused by an intraoperative embolic occlusion of the anterior spinal artery. Cruciate paralysis and man-in-barrel syndrome (MIBS) both are terms used to describe brachial diplegia; cruciate paralysis when caused by medullary lesions, MIBS when caused either by supratentorial or by medullary lesions. Exclusive use of the term MIBS for bilateral frontal lobe lesions, as in the original description, would provide more clarity in terminology. PMID- 11939952 TI - Syringomyelia with syringobulbia presenting only with paralysis of 9th and 10th cranial nerves. AB - We report a case of syringomyelia with syringobulbia, in a patient who presented only with involvement of the 9th and 10th cranial nerves. PMID- 11939953 TI - Post-epidural analgesia spinal cord infarction: MRI correlation. AB - Permanent neurological deficits after epidural analgesia are rare, but have long been believed to be caused by cord ischaemia when no obvious cause is demonstrable. The mechanisms of this injury are uncertain, but a literature review suggests important risk factors. We report a first case of extensive spinal cord infarction confirmed by magnetic resonance imaging (MRI) following post-thoracotomy epidural analgesia and review the literature to explain the mechanism underlying this devastating complication. PMID- 11939954 TI - "Restless legs" in "The Wedding Proposal" by Anton Chekhov. PMID- 11939955 TI - Paraneoplastic subacute encephalitis caused by adenocarcinoma of prostate: a clinico-pathological case report. PMID- 11939956 TI - Serum cholesterol concentration, depression, and anxiety. PMID- 11939957 TI - Social phobia: overview of community surveys. AB - OBJECTIVE: Social phobia has in recent years been recognized as a considerable public health concern. This paper presents an overview of community studies that have estimated the prevalence of social phobia, its subtypes, and specific social fears in the adult general population. Sociodemographic findings are also summarized. METHOD: Forty-three epidemiological studies from 1980 to the present were located in electronic databases and by complementary searches of citations in journal articles and book chapters. RESULTS: Recent surveys estimate the lifetime prevalence of social phobia at about 7-13% in Western countries. However, prevalence rates vary considerably and are prone to several influencing methodological variables such as the diagnostic criteria, diagnostic threshold, assessment method, number of situational probes, and prevalence period selected by the researcher. CONCLUSION: Although there are difficulties in delineating social phobia from subsyndromal social anxiety or shyness, social phobia is even when narrowly defined remarkably common in the general population. PMID- 11939958 TI - Personality dimensions in social phobics with or without depression. AB - OBJECTIVE: This study investigated the personality traits of social phobics using the Temperament and Character Inventory (TCI). METHOD: A sample of 178 social phobics was assessed with the TCI, and compared with controls. The patients were classified into two groups, according to the absence (SP group) or to the presence (SP+D group) of depression. RESULTS: We found significant elevated scores for harm avoidance (HA) in social phobics when compared with controls (16.2 +/- 2.7), in both the SP (26.2 +/- 3.5), and the SP+D (28.9 +/- 4.7), groups. Lower self-directedness scores were found in the SP and in the SP+D groups when compared with the controls. Patients with the generalized type of social phobia had higher HA scores as compared with other social phobics. CONCLUSION: The personality profile obtained in these social phobics, whatever their depressive symptomatology, reflects a dramatically anxious and avoidant temperament associated to an immature character. PMID- 11939960 TI - Risk factors in the early family life of patients suffering from dissociative disorders. AB - OBJECTIVE: To test the hypothesis that not only intrafamiliar childhood abuses inflicted to the patient, but also major losses or other severe life events suffered by the mother within 2 years of patient's birth, are risk factors for the development of dissociative disorders (DDs). METHOD: A multicentric case control study of 52 cases and 146 controls. RESULTS: We estimated a crude odds ratio (OR) of 2.6 (adjusted 1.9) for mothers' losses or other severe life events experienced within 2 years of patient's birth, and a crude OR of 7.6 (adjusted 7.2) for patient's early traumatic experiences. CONCLUSION: Mother's losses or other severe life events within 2 years of patient's birth and patient's traumatic experiences during childhood are risk factors for the development of dissociative disorders. A possible explanation of these findings is that disorganized or insecure attachment may increase susceptibility to traumatic experiences and propensity to dissociation in adult life. PMID- 11939959 TI - Acute polymorphic psychotic disorder: diagnostic stability over 3 years. AB - OBJECTIVE: Acute polymorphic psychotic disorder without symptoms of schizophrenia (APPD) in ICD-10 is unique in many characteristics. This study aimed at investigating the diagnostic stability of APPD over 3 years. METHOD: Forty-five patients with first episode APPD were followed up and assessed at regular intervals over a period of 3 years. RESULTS: Thirty-two patients were females. Average age of the sample was 26.9 years. Thirty-three cases retained their index diagnosis of APPD, while 12 cases required diagnostic revision: 10 to bipolar affective disorder and the rest to unspecified non-organic psychosis. Shorter duration of illness (<1 month) and abrupt onset (<48 h) predicted a stable diagnosis of APPD. CONCLUSION: APPD is a relatively stable diagnosis and argues for a distinct diagnostic entity. PMID- 11939961 TI - Referrals to a liaison psychiatry out-patient clinic in a UK general hospital: a report on 900 cases. AB - OBJECTIVE: To describe the demographic and clinical characteristics of consecutive referrals to an out-patient liaison psychiatry clinic in a large university hospital in the UK. METHOD: We studied 900 patients using a standardized proforma. Assessments of functional capacity were made using the Global Assessment of Functioning (GAF) scale, and each patient was assigned a psychiatric diagnosis using ICD-10 criteria. RESULTS: Four of five of all referrals presented with somatic complaints, and 41% had a concurrent physical illness. The most common psychiatric diagnoses were somatoform disorders (55%) and neurasthenia (11%). Although more than half (59%) had no previous psychiatric history, a surprisingly high number (35%) had significant functional impairment (scores of < 50 on the GAF scale). The lack of adequate psychological treatment services often provided a barrier to optimal management of some of the more disabled patients. CONCLUSION: The general hospital liaison psychiatry clinic provides an acceptable setting in which to assess and manage patients referred from non-psychiatric colleagues. PMID- 11939962 TI - Bias in computerized neuropsychological assessment of depressive disorders caused by computer attitude. AB - OBJECTIVE: Psychiatric patients are increasingly confronted to computerized psychological and psychopathological assessment. Patients' attitude to computers was reported to affect acceptance of computerized assessment. METHOD: In 78 psychiatric in-patients neuropsychological impairment was examined following admission on an open ward by conventional as well as computerized memory and attention tasks. Besides psychopathological assessment, self ratings of computer attitude and acceptance of the computerized assessment were completed. RESULTS: A more negative attitude to computers was found to be significantly correlated to higher nervousness in patients' self report (R=0.38, P=0.0005) as well as to poorer results of computerized attention tasks (R=0.39, P=0.0007). Particularly in patients with depressive disorders computer attitude could be shown to explain 39% of the variance of attention performance. CONCLUSION: Results indicate a significant effect of negative computer attitude on acceptance and thus reliability of computerized examination, resulting in a bias in computerized attention-related assessment in patients with depressive disorders. PMID- 11939963 TI - Determinants of subjective quality of life in patients attending community-based mental health services. The South-Verona Outcome Project 5. AB - OBJECTIVE: Investigate the relationship of various domains of quality of life (QoL) with socio-demographics, clinical and social characteristics, service use and satisfaction in a representative sample of patients in contact with the South Verona community mental health service. METHOD: Measures included: Lancashire Quality of Life Profile (LQOLP), demographics, diagnosis and service utilization data from the Case Register, Brief Psychiatric Rating Scale (BPRS), Disability Assessment Schedule (DAS), Global Assessment of Functioning (GAF) and Verona Service Satisfaction Scale (VSSS). Analyses were conducted using a block stratified multiple regression model. RESULTS: Demographics, diagnosis, psychopathology, disability, functioning and service use together explained different amounts of variance in each LQOLP domain, but always less than 14%. VSSS explained more variance than any other indicator in health (13.1%), social relations (12.1%), leisure/participation (9.1%), and general wellbeing (9.0%). CONCLUSION: Different domains of QoL are predicted by different indicators. In some important domains, self-perceived satisfactory and effective care might have an impact on the QoL of patients. PMID- 11939964 TI - Serum cholesterol and psychological distress in hospitalized depressed patients. AB - OBJECTIVE: To assess the relationship between total serum cholesterol and various psychosocial variables in depressed in-patients. METHOD: One hundred and eighty six patients had their total fasting serum cholesterol assessed following admission; psychiatric diagnoses were obtained with the structured clinical interview for DSM-IV (SCID) interview. Psychopathology was measured with a clinician rated scale [Montgomery and Asberg Depressive Rating Scale (MADRS)] and a self-rating scale [Symptom checklist-90 (SCL-90)]. RESULTS: Univariate analyses showed lower total serum cholesterol levels being correlated with higher scores in several psychopathological areas. Multivariate analyses indicated that male gender, lower age and higher MADRS scores were the most predictive variables for lower cholesterol levels. CONCLUSION: The data suggest, in this depressed population, an association between serum cholesterol and depressive symptoms. What is the cause and what is its effect is not possible to say from this cross sectional study. PMID- 11939968 TI - Non-adherence in psychiatric disorders: misbehaviour or clinical feature? PMID- 11939969 TI - Treatment non-adherence in affective disorders. AB - OBJECTIVE: The aim of this paper is to review the prevalence, predictors and methods for improving medication adherence in unipolar and bipolar affective disorders. METHOD: Studies were identified through Medline and PsycLit searches of English language publications between 1976 and 2001. This was supplemented by a hand search and the inclusion of selected descriptive articles on good clinical practice. RESULTS: Estimates of medication non-adherence for unipolar and bipolar disorders range from 10 to 60% (median 40%). This prevalence has not changed significantly with the introduction of new medications. There is evidence that attitudes and beliefs are at least as important as side-effects in predicting adherence. The limited number of empirical studies of how to reduce non-adherence offer encouraging evidence that, if recognized, the problem can be overcome. CONCLUSION: Only 1-2% of all publications on the treatment of affective disorders explore factors associated with medication non-adherence. This is disappointing as research and clinical data highlight the importance of extended courses of medication in improving the long-term prognosis of affective disorders. PMID- 11939970 TI - Cannabis-induced psychosis: a cross-sectional comparison with acute schizophrenia. AB - OBJECTIVE: The existence of cannabis-induced psychosis (CP) remains controversial, partly because of methodological problems. We hypothesize that acute schizophrenia (AS) and CP can have distinct demographic, premorbid and clinical features. METHOD: We compared 26 patients with CP to 35 with AS, after their cannabis-consumption status was confirmed by repeated urine screens. Patients with CP were assessed after at least 1 week but not more than 1 month of abstinence. Symptoms were evaluated with the Present State Examination (PSE). RESULTS: In group CP, male gender, expansive mood and ideation, derealization/depersonalization, visual hallucinations, and disturbances of sensorium were more frequent than in group AS. Premorbid schizoid personality traits were more frequently associated to AS and antisocial personality traits to CP. CONCLUSION: The continuous heavy use of cannabis can induce a psychotic disorder distinct from AS. These two clinical entities share some features but they differ in others. PMID- 11939971 TI - Obstetrical complications, parenting practices and risk of criminal behaviour among persons who develop major mental disorders. AB - OBJECTIVE: The present study examined whether socio-economic status (SES) of the family of origin, inadequate parenting, and pre and perinatal factors are antecedents of criminality among men and women who develop a major mental disorder. METHOD: The sample included the 82 men and 79 women from the 1953 Stockholm birth cohort who developed a major mental disorder by age 30. Information was extracted from obstetric files, health, social and work records and official criminal records. RESULTS: Among males, neonatal complications increased the risk of offending two-fold, the risk of violent offending 2.5 times, and the risk of early start offending 3 times. Neither pregnancy or birth complications, inadequate parenting, or SES of the family of origin increased the risk for offending. Among females, none of the factors that were examined were associated with offending. CONCLUSION: Complications in the neonatal period are associated with offending among men who develop major mental disorders. PMID- 11939972 TI - Clinical issues related to depression in schizophrenia: an international survey of psychiatrists. AB - OBJECTIVE: Depressive symptoms in schizophrenia are associated with a substantial morbidity and mortality burden. The 'International Survey of Depression in Schizophrenia' was designed to evaluate current awareness and clinical approaches in this area. METHOD: A 48-item questionnaire was distributed to approximately 80 000 consultant psychiatrists world-wide. Responses were analysed using descriptive statistics. RESULTS: Respondents demonstrated considerable awareness of the prevalence and consequences of depression in schizophrenia. Although there was widespread adjunctive use of antidepressants, one-third of respondents indicated that they rarely or never prescribe these agents in combination with antipsychotic medication. There were considerable variations in opinions about the best approach to the treatment of depressive symptoms associated with schizophrenia. CONCLUSION: The considerable clinical burden of depressive symptomatology in schizophrenia was acknowledged by the majority of respondents to this survey. There was, however, little agreement on the best management strategy. PMID- 11939973 TI - The influence of neurocognitive deficits and symptoms on disability in schizophrenia. AB - OBJECTIVE: The purpose of the present study was to examine the relationship of disability with neurocognitive deficits and symptoms in schizophrenia. METHOD: Sixty patients with schizophrenia and 30 healthy controls matched for age, sex and level of education were included in the study. Neurocognitive tests measuring attention, visual memory and executive functions were given. Severity of symptomatology was assessed by the Positive and Negative Syndrome Scale (PANSS). Disability level of the subjects was assessed by World Health Organisation Disability Assessment Schedule 2 (WHO-DAS-2). RESULTS: PANSS total score and the subscores were all correlated with DAS scores at a significant level. Neurocognitive test scores were not significantly associated with disability level. Regression analysis furthermore showed that symptom severity was predictive of the disability level. CONCLUSION: These results suggest that, rather than neurocognitive deficits, symptoms appear to have direct impact on the functioning of patients with schizophrenia in many domains of life. PMID- 11939974 TI - Could the hypofrontality pattern in schizophrenia be modified through neuropsychological rehabilitation? AB - OBJECTIVE: The effects of neuropsychological treatment on cognitive hypofrontality were examined in schizophrenic patients through the score activation. METHOD: Eight subjects (six men and two women) with persistent negative symptoms and cognitive impairments were evaluated with single photon emission computed tomography (SPECT) procedures and neuropsychological battery before and after a neuropsychological treatment group. RESULTS: After treatment an enhancement in neuropsychological performance was found, especially in executive functions. The activation score showed an increase over baseline levels and no cognitive-dependent hypofrontality after treatment was found. Although the prefrontal blood flow changes were small and non-specific, they suggest a reduction of the cognitive hypofrontality after neuropsychological treatment. CONCLUSION: Cognitive improvements after neuropsychological treatment would possibly be related with the diminution of the functional hypoactivity in the prefrontal areas. PMID- 11939975 TI - Demographic, family, and occupational characteristics associated with major depression: the Harvard study of moods and cycles. AB - OBJECTIVE: This study assesses the extent to which women with and without major depression differ by demographic, familial, and occupational characteristics. METHOD: From a community-based sample, the authors identified 332 women with and 644 women without current or past major depression based on Structured Clinical Interviews for DSM-IV. Demographic and background interviews were conducted in person. RESULTS: Depressed women were more likely to have gained >or =35 lbs between age 18 and study enrollment (OR=1.6, 95% CI 1.1-2.5), experienced divorce (OR=2.0, 95% CI 1.4-2.8), or changed occupations (OR=1.5, 95% CI 1.1-2.1) compared with non-depressed women. Compared with women with no brothers, those with > or =1 brothers were less likely to have a history of depression (OR=0.8, 95% CI 0.6-1.1), whereas compared with women with no sisters, those with > or =1 sisters were more likely to have current or past depression (OR=1.4, 95% CI 1.0 1.9). These findings were not influenced by family sibship size. CONCLUSION: These results illustrate demographic differences between women with and without major depression and that sibship gender rather than size may also influence risk. PMID- 11939976 TI - The prospectively observed course of illness among depressed patients who commit suicide. AB - OBJECTIVE: These analyses were conducted to describe the course of illness among patients with major affective disorders who commit suicide. METHOD: Twenty-nine patients who entered a long-term, high-intensity follow-up study of major affective disorders and who later committed suicide within 1 year of their last follow-up interview were individually matched to other patients by age, sex, the presence or absence of lifetime drug or alcohol abuse, time to last interview and polarity. Those who suicided were compared with their controls by depressive and substance abuse morbidity during follow-up, treatment resistance, treatment compliance, suicidal behavior and psychosocial adjustment. RESULTS: Among the various measures used to characterize the course of illness during a mean follow up of 4.3 years, only those pertaining to suicidal behavior robustly separated the suicide group from their controls. Suicidal behavior in the remote past seemed as predictively important as suicidal behavior during follow-up. CONCLUSION: Of the various features monitored over time in patients with major affective disorder, suicidal behavior itself was the clearest correlate of risk for completed suicide. PMID- 11939977 TI - Psychiatric symptoms in neurologically asymptomatic Huntington's disease gene carriers: a comparison with gene negative at risk subjects. AB - OBJECTIVE: Psychiatric profiles of two at-risk groups [Huntington's disease (HD) gene carriers and non-carriers] were compared by means of a computerized battery and a structured interview. METHOD: To avoid confounding, only subjects who were free from neurological and cognitive deficits (neurologically asymptomatic) were included in the study. To avoid evaluation biases, all subjects were seen before the genetic testing was undertaken. RESULTS: Gene carriers had significantly worse recognition memory and scored higher in measures of irritability than controls. The groups also differed in terms of the factor structure of their psychiatric symptoms. None of the subjects qualified for a psychiatric diagnosis at the time of assessment. CONCLUSION: The groups differed with respect to their profile of psychiatric symptoms. It is hypothesized that these differences are the expression of different mechanisms, i.e. that cognitive deficits relate more to genetic factors and neurotic complaints more to being brought up in a disturbed family background. Issues concerning instrument sensitivity, selection bias and the advantage of seriatim assessments are discussed. PMID- 11939978 TI - Improvement in activities of daily living in elderly following treatment for post bereavement depression. AB - OBJECTIVE: To determine if elderly bereaved depressed subjects display difficulty with activities of daily living (ADL) and if their ADL difficulty improves with psychopharmacologic intervention. METHOD: The Assessment of Motor and Process Skills (AMPS), an ADL assessment measuring ADL motor and ADL process skills, was administered to a pilot sample of elderly persons with post-bereavement depression prior to psychopharmacologic intervention and subsequently during treatment response. RESULTS: In the pretreatment phase, subjects displayed difficulty with ADL motor and ADL process skills that significantly improved during the treatment response phase. CONCLUSION: Results suggest that elderly bereaved depressed individuals demonstrated ADL difficulty that responds positively to psychopharmacologic intervention. PMID- 11939979 TI - Olanzapine-induced destabilization of diabetes in the absence of weight gain. AB - OBJECTIVE: To provide evidence that olanzapine can cause glucose dysregulation by a mechanism other than weight gain. METHOD: I report a case of a diabetic patient who developed glucose dysregulation soon after initiation of olanzapine treatment, occurring in the absence of weight gain. I compare this case to previous case reports. RESULTS: Our patient developed persistent hyperglycaemia within 3 weeks of initiating treatment with olanzapine. Weight recorded just prior to commencement and soon after discontinuation of olanzapine were not significantly different. CONCLUSION: Controlled studies are necessary to elucidate the mechanism by which olanzapine can cause dysregulation of glucose homeostasis, and to develop guidelines for the use of olanzapine in patients with known diabetes as well as in patients with risk factors for diabetes. PMID- 11939987 TI - Up, up and away: watching the Caesarean section rate rise. PMID- 11939988 TI - Osteocalcin and the hormonal, inflammatory and metabolic response to major orthopaedic surgery. AB - Plasma osteocalcin, a marker of osteoblastic activity, decreases after major abdominal and gynaecological surgery. Increased cortisol secretion and other hormonal and inflammatory components of the peri-operative stress response may play a role in mediating this response. We assessed the effects of three different anaesthetic techniques on peri-operative osteocalcin concentrations. Thirty-six female patients undergoing elective total hip replacement were randomly assigned to receive propofol, propofol plus 'three-in-one' block or etomidate as part of a general anaesthetic technique. We measured plasma osteocalcin and serum cortisol, bone specific alkaline phosphatase, interleukin 6, plasma epinephrine, norepinephrine, plasma glucose and cystatin C concentrations for up to 3 days after surgery. Etomidate successfully inhibited the cortisol response to surgery but plasma osteocalcin declined in all patients. This was accompanied by increased plasma catecholamines, interleukin-6 and glucose concentrations, and decreased cystatin C-values. Inhibition of the cortisol response to surgery failed to prevent a decrease in plasma osteocalcin concentrations after surgery, suggesting that other factors such as cytokines or catecholamines may play a significant role. PMID- 11939989 TI - A novel method of measuring cricoid force. AB - The application of cricoid pressure is an effective means of preventing regurgitation of gastric contents when correctly applied. A force of 30 N (3 kg) is recommended for an unconscious patient. This study assesses the validity of using floor scales to measure cricoid force applied by anaesthetic assistants and compares their use to an established training technique. Forty subjects applied pressure to a cricoid model in a blinded manner, on three test occasions. For each test, cricoid pressure was maintained for 1 min and the highest and lowest forces recorded on the model were noted. The first test was before any instruction. The second test followed a period of practice on the cricoid model. For the final test, subjects stood upon a set of floor scales and noted their weight. Force was applied to the cricoid model until the weight on the floor scales reduced by 3 kg. Performance improved both following practice on the model and using the floor scales. Applying cricoid force while standing on floor scales and using the change in weight as a guide resulted in a predictable force on the cricoid model. The use of floor scales is a useful method of demonstrating the forces needed for effective cricoid pressure. PMID- 11939990 TI - The cardiovascular response to insertion of the intubating laryngeal mask airway. AB - Sixty-one patients received a standardised anaesthetic and were randomly assigned to three groups: tracheal intubation via direct laryngoscopy, tracheal intubation via an intubating laryngeal mask airway with immediate removal of the device, and tracheal intubation via an intubating laryngeal mask airway with delayed removal. The cardiovascular response to intubation was of a similar magnitude in all groups, although delayed removal of the intubating laryngeal mask airway was associated with a second pressor response. Norepinephrine changed significantly over time following direct laryngoscopy and following immediate removal of the intubating laryngeal mask airway, but not after delayed removal. The findings of this study do not support using the intubating laryngeal mask instead of direct laryngoscopy purely to decrease the response to intubation. PMID- 11939991 TI - A retrospective observational study of pre-operative sickle cell screening. AB - The aim of this study was to determine the ethnic mix of those patients being pre operatively screened for sickle cell disease in a London teaching hospital and to determine the rate of carriage of sickle haemoglobin amongst those tested. We retrospectively studied 1879 patients undergoing surgery over a 2-month period. Two hundred and thirteen (11%) were screened for sickle cell disease and of these, 12 (5%) tested positive for sickle cell trait (HbAS). There were no patients homozygous for sickle cell disease (HbSS) or with haemoglobin SC disease (HbSC). Screening rates varied widely in different ethnic groups from 0% of the Chinese population to 85.2% of the Afro-Caribbean population. We conclude that at present there is no coherent pre-operative screening policy for sickle cell disease in our institution. Sickle cell disease poses unique anaesthetic risks and with a rapidly expanding 'mixed race' population high-risk patients are difficult to identify phenotypically. We propose a universal screening policy be implemented in high-risk areas. PMID- 11939992 TI - Isoflurane and coronary heart disease. AB - Early studies indicated that isoflurane caused coronary steal and should therefore be avoided in patients with coronary heart disease. Subsequently, more detailed trials have disputed this and have shown that as long as coronary perfusion pressure is maintained, isoflurane does not cause coronary steal or myocardial ischaemia. There is now growing evidence, initially in animal work but more recently in human studies, that isoflurane has myocardial protective properties, limiting infarct size and improving functional recovery from myocardial ischaemia. The mechanism for this protection mimics ischaemic preconditioning and involves the opening of adenosine triphosphate-dependent potassium channels. The few studies comparing the myocardial protection offered by individual anaesthetic agents indicate that isoflurane represents the anaesthetic agent of choice for patients with coronary heart disease. PMID- 11939993 TI - Acid-base physiology: the 'traditional' and the 'modern' approaches. AB - The interpretation and understanding of acid-base dysfunction has recently been revisited. The 'traditional' approach developed from the pioneering work of Henderson and Hasselbalch and is still the most widely used in clinical practice. There are a number of problems identified with this approach, however. The 'modern' approach derives from Stewart's work in physical chemistry. In this review we describe the origins of the traditional approach and discusses related concepts. We then describe Stewart's approach, including how it is derived and how it may be used to classify acid-base derangements. The applications of Stewart's approach to clinical scenarios in intensive care is then discussed briefly before we examine some published clinical studies based on his work. PMID- 11939994 TI - Sir Humphry Davy; his researches in respiratory physiology and his debt to Antoine Lavoisier. AB - This article shows how the original works of the French scientist Antoine Lavoisier were developed by Humphry Davy, a trainee surgeon from Cornwall, while he was working as a physiologist. Antoine Lavoisier had worked out how oxidation involved the consumption of oxygen and the release of energy. Davy's book, Researches Chemical and Philosophical, Chiefly Concerning Nitrous Oxide, published in 1799, describes the measurement of his own lung volumes, including the first recorded measurement of the residual volume. He measured his own rates of oxygen consumption and carbon dioxide production. He is famous for his investigations into nitrous oxide, but he also investigated the effects of breathing nitric oxide and carbon monoxide. He made these observations with a gasometer and analysis of his expired air, and his work anticipates the invention of blood gas analysis. PMID- 11939995 TI - A survey of the use of portable ultrasound for central vein cannulation on critical care units in the UK. AB - A questionnaire was sent to 288 critical care units in the UK to assess the use of portable ultrasound machines to assist central vein cannulation. There was a 58% response rate. Ultrasound guidance was used by 36 (21.6%) units for central vein cannulation. Of these, only four (11.1%) used it routinely and 25 (69.4%) used it when faced with a difficult vein cannulation. Half of the units with ultrasound facilities may be using it suboptimally. Of those units not using ultrasound for central vein cannulation, 70 (53%) said it was because of lack of equipment and 51 (38.9%) did not think that it was necessary. Overall, over half of the units did not audit complications of central vein cannulation. PMID- 11939996 TI - Pump activated by a foot switch pedal for controlled administration of local anaesthetic drugs. AB - We describe a newly designed syringe pump which is electrically controlled by a dual foot switch pedal. The device enables the scrubbed anaesthetist performing the regional block to aspirate, as well as to inject, the local anaesthetic without the need for any additional personnel. PMID- 11939997 TI - Pressure in the cuffs of tracheal tubes at altitude. AB - Pressures in the cuffs of three commonly used tracheal tubes (Portex Profile Softseal, Mallinckrodt Lo-Contour and Mallinckrodt Hi-Contour, size 8.0 mm and 9.0 mm internal diameter), inflated with air, were measured during simulated ascents in an altitude chamber to 10 000 ft. There was no detectable difference in performance between sizes for each type of tracheal tube. When averaged over the two sizes for each type of tube, cuff pressure reached the critical perfusion pressure 50 cmH2O (37 mmHg) for tracheal mucosa at a higher altitude in the Portex Profile Softseal (2837 ft, 95% CI 2488-3186 ft) than in the Mallinckrodt Lo-Contour (2128 ft, 95% CI 1779-2476 ft; p = 0.02) and Mallinckrodt Hi-Contour (1820 ft; 95% CI 1471-2168 ft; p = 0.002) tracheal tubes. When the cuffs of the 9.0-mm tracheal tubes were inflated with saline, much smaller increases in pressure were measured with increasing altitude, although inflation of the cuffs with saline was technically difficult. Commonly used tracheal tubes with air inflated cuffs can be used for aeromedical retrieval, but air should be evacuated from the cuffs after increases in altitude of as little as 2000-3000 ft. PMID- 11939998 TI - Survey of the use of the gum elastic bougie in clinical practice. AB - Data were collected prospectively on the use of the gum elastic bougie in 200 patients. The bougie was successfully inserted into the trachea and tracheal intubation was accomplished in 199 cases. The bougie was inserted into the trachea at the first attempt in 178 cases. In nine cases (4.5%) a second, more experienced, clinician was required. In 173 cases, the grades of view were recorded before and after the application of laryngeal pressure; pressure improved the view in 80 cases (46%), had no effect in 89 (51%) and worsened the view in four cases (2%). Various recommendations for optimal external laryngeal pressure and use of the bougie were not followed on 15-64% of occasions. There is a need for better education in these techniques. PMID- 11939999 TI - A combined technique utilising regional anaesthesia and target-controlled sedation in a patient with myotonic dystrophy. AB - Myotonic dystrophy presents several problems to the anaesthetist. We describe what we believe to be the first report of target-controlled sedation combined with regional anaesthesia in a patient with myotonic dystrophy. Precise control of propofol levels and titration to patient satisfaction avoided the problem of delayed recovery which has been described with propofol anaesthesia. PMID- 11940000 TI - The acute pain service, a model for outreach critical care. PMID- 11940001 TI - The intubating laryngeal mask airway as a guide to fibreoscopy. PMID- 11940002 TI - Sniff the morning air or drink a pint of beer? PMID- 11940003 TI - Switching on to ultrasound. PMID- 11940004 TI - Nasal insertion of the oesophageal Doppler probe. PMID- 11940005 TI - Reflection; end of an era and a new beginning! PMID- 11940006 TI - Etomidate, adrenal dysfunction and critical care. PMID- 11940007 TI - Alpha-agonists use in cardiac arrest. PMID- 11940008 TI - Use of thiopental for rapid sequence induction. PMID- 11940010 TI - Midazolam--an anti-emetic? PMID- 11940015 TI - Totally implantable total artificial heart for clinical application. PMID- 11940016 TI - Denervated muscles in humans: limitations and problems of currently used functional electrical stimulation training protocols. AB - Prior clinical work showed that electrical stimulation therapy with exponential current is able to slow down atrophy and maintain the muscle during nonpermanent flaccid paralysis. However, exponential currents are not sufficient for long-term therapy of denervated degenerated muscles (DDMs). We initiated a European research project investigating the rehabilitation strategies in humans, but also studying the underlying basic scientific knowledge of muscle regeneration from satellite cells or myoblast activity in animal experiments. In our prior study, we were able to show that high-intensity stimulation of DDMs is possible. At the beginning of training, only single muscle twitches can be elicited by biphasic pulses with durations of 120-150 ms. Later, tetanic contraction of the muscle with special stimulation parameters (pulse duration of 30-50 ms, stimulation frequency of 16-25 Hz, pulse amplitudes of up to 250 mA) can improve the structural and metabolic state of the DDMs. Because there are no nerve endings for conduction of stimuli, large-size, anatomically shaped electrodes are used. This ensures an even contraction of the whole muscle. Contrary to the current clinical knowledge, we were able to stimulate and train denervated muscle 15-20 years after denervation. The estimated amount of muscle fibers that have to be restored is about 2-4 million fibers in each m. quadriceps. To rebuild such a large number of muscle fibers takes up to 3-4 years. Despite constant stimulation parameters and training protocols, there is a high variation in the developed contraction force and fatigue resistance of the muscle during the first years of functional electrical stimulation. PMID- 11940017 TI - Transcutaneous functional electrical stimulator "Compex Motion". AB - Research groups in the field of functional electrical stimulation (FES) are often confronted with the fact that existing and commercially available FES stimulators do not provide sufficient flexibility and cannot be used to perform different FES tasks. The lack of flexibility of the commercial systems until now forced various FES research teams to develop their own stimulators. This paper presents a newly developed firmware and graphical programming software for the commercial Compex 2 stimulator which enhances the versatility and capabilities of the stimulator from a medical and therapeutic device to a neuroprosthesis and research tool. The new stimulator, called Compex Motion, can now be used to develop various custom-made neuroprostheses, neurological assessment devices, muscle exercise systems, and experimental setups for physiological studies. It can be programmed to generate any arbitrary stimulation sequence that can be controlled or regulated by various external sensors, sensory systems, or laboratory equipment. By interconnecting two or more Compex Motion stimulators, the number of stimulation channels can be increased to multiples of four channels, 8, 12, 16, 20, and so forth. The stimulation sequences and the control strategies are programmed and stored on exchangeable credit card-sized memory chip cards. The stimulator has four biphasic current-regulated stimulation channels and two general purpose analog input channels that can be configured to measure the output voltage of a variety of sensors such as goniometers, inclinometers, gyroscopes, or electromyographic (EMG) sensors. For real-time EMG control of the stimulation patterns, an EMG processing algorithm with software stimulation artifact blanking was implemented. The Compex Motion stimulator is manufactured by the Swiss company Compex SA and is currently undergoing clinical trials. PMID- 11940018 TI - The Vienna functional electrical stimulation system for restoration of walking functions in spastic paraplegia. AB - An eight-channel stimulation system, currently intended for stimulation of lower extremities, was developed and is introduced. The major development goals were easy handling, modularity to make the system easily adaptable for other functional electrical stimulation (FES) applications, and a wide stimulation parameter range for application-specific parameter optimization. For paraplegic stepping, the system worn by the patient consists of 2 four-channel stimulation modules, a central unit holding the battery and circuitry for power management and communication control, a wireless remote control unit, and a palmtop computer as the main control and input device. A software package for Microsoft Windows supports the design and optimization of stimulation sequences in the rehabilitation center. First tests with patients familiar with FES showed smoother movements during stepping and acceptable good handling. In combination with the PC software, the required stimulation sequences could be created in a very short time. PMID- 11940019 TI - Second-generation microstimulator. AB - The first-generation injectable microstimulator was glass encased with an external tantalum capacitor electrode. This second-generation device uses a hermetically sealed ceramic case with platinum electrodes. Zener diodes protect the electronics from defibrillation shocks and from electrostatic discharge. The capacitor is sealed inside the case so that it cannot be inadvertently damaged by surgical instruments. This microstimulator, referred to as BION, is the main component of a 255-channel wireless stimulating system. BION devices have been implanted in rats for periods of up to 5 months. Results show benign tissue reactions resulting in identical encapsulation around BION and controls. Stimulation threshold levels did not change significantly over time and ranged between 0.81 to 1.35 mA for all the animals at a 60 micros pulse width. All of the tests performed to date indicate that the BION is safe and effective for long term human implant. We have elected to develop BION applications by seeking collaboration with the research community through our BION Technology Partnership. PMID- 11940020 TI - The microsystems based visual prosthesis for optic nerve stimulation. AB - The microsystems based visual prosthesis (MiViP) visual prosthesis generates visual perceptions well below safety and stimulator saturation limits. These perceptions, called phosphenes, are of reasonably small size and are broadly distributed in the visual field. They can thus be used to convey useful visual information. Psychophysical evaluations are being performed in order to assess the implantee's benefits in the use of the MiViP optic nerve visual prosthesis. In a pattern-recognition task, the performance improved regularly with practice with an increasing score and a decreasing delay to recognition. These observations open the way toward an evaluation of general mobility improvement with the portable system. In conclusion, the results obtained so far still support the potential usefulness of the optic nerve visual prosthesis. A low resolution artificial vision can be expected from the prosthesis after extensive training. PMID- 11940021 TI - The hemodynamic function of intrathoracic skeletal muscle ventricles after recovery from surgery in pigs. AB - The shortage of donor organs for heart transplantation highlights the need for new approaches to end-stage heart failure. A promising experimental technique is the use of pumping chambers formed from the latissimus dorsi muscle. We formed such skeletal muscle ventricles (SMVs) and connected them to the descending thoracic aorta in a single surgical procedure in pigs. Activation of conditioned SMVs from the end of systole for 80% of diastole increased mean aortic diastolic blood pressure by 11.2 +/- 1.6% in 1 animal and by 15.8 +/- 0.3% in another. The left-ventricular stroke work in the postassisted beat was decreased by 8.7 +/- 5.8% and 10.1 +/- 2.2% and the overall stroke work by 7.4 +/- 1.2% and 9.4 +/- 0.8%. The key to forming and connecting the SMV in a single procedure was the use of a composite homograft lining. In future clinical practice, this component could be replaced by a synthetic composite or by a tissue lining produced in vitro. PMID- 11940022 TI - Development of muscular blood pumps performed in a one-step operation. AB - Up to now, the employment of skeletal muscle ventricles (SMVs) has required a two step operation, that is, first, the construction followed by a vascular delay and electrical conditioning, and second, the integration into circulation by a second operation. As shown previously, clenbuterol increased the power of electrically conditioned SMVs wrapped around a mock system. These clenbuterol-supported SMVs pumped successfully from construction to several months against a pressure of 60 70 mm Hg. Due to these successful former experiments, a muscular blood pump has been employed via a one-step procedure, trained within the circulation under the support of clenbuterol. It appeared to be hemodynamically relevant and is expected to become clinically practicable for the treatment of end-stage heart failure. PMID- 11940023 TI - Stimulation of nerves innervating the dog's pancreas. AB - Our aim was to modulate secretion of insulin and glucagon into the blood of a diabetic but otherwise healthy dog with stimulation of nerves innervating the pancreas. The 33 electrode spiral cuffs were implanted in an adult beagle canine. The first cuff was installed on the vagus nerve, the second one on the splanchnic nerve, and the last one on the pancreatic nerve. To cause insulin-dependent Type I diabetes, partial dysfunction of the pancreas was induced. Nerves were stimulated using biphasic, rectangular current pulses (10 mA, 200 micros, 20 Hz). Samples from the femoral artery were drawn before the experiment, after 5 min, and 5 min after the stimulation stopped. The results of radioimmunological assay of blood samples showed that in the intact pancreas, stimulation of the vagus nerve caused a considerable increase in insulin secretion, no significant change in glucagon secretion, and a decrease in C-peptide secretion. Splanchnic nerve stimulation did not cause considerable change in insulin and C-peptide secretion while considerable increase in glucagon secretion was noticed. Pancreatic nerve stimulation did not considerably change the secretion of any of the three hormones. In the dysfunctioned pancreas, vagal nerve stimulation caused an increase in insulin and glucagon secretion and a minor increase in C-peptide secretion. Splanchnic nerve stimulation caused a minor decrease in insulin secretion, a considerable increase in glucagon secretion, and a small increase in C-peptide secretion. Pancreatic nerve stimulation did not cause a considerable change in insulin secretion while a minor increase in glucagon and C-peptide secretion was observed. PMID- 11940024 TI - Functional electrical stimulation-induced surface muscle stiffness captured by computer-controlled tonometry. AB - A new tonometric test system to assess surface stiffness over relaxed and activated calf muscles was developed. The mechanical arrangement consists of a skin indentor driven by a torque motor (galvo-drive) that is rigidly connected to an ankle dynamometer. The indentation depth is measured by a displacement transducer. Software routines for cyclic indentation (recording of stiffness curves), static indentation (sensing of twitch responses), and vibration (skin resonance) were implemented. A visual interface is used to capture surface stiffness during target contractions and during controlled relaxation. For functional electrical stimulation (FES) applications, the software includes a pulse train synthesizer to generate arbitrary stimulation test patterns. The system's performance was tested in FES and voluntary contraction procedures. PMID- 11940025 TI - Devices for noninvasive transcranial electrostimulation of the brain endorphinergic system: application for improvement of human psycho-physiological status. AB - It is well known that deficit of endorphins plays an important role in disturbances of human psycho-physiological status. Previously, we revealed that brain endorphinergic structures have quasiresonance characteristics. On the basis of these data, a method of activation of the brain endorphinergic structures by means of noninvasive and rather selective transcranial electrostimulation (TES) as a kind of functional electrical stimulation (FES) was elaborated. New models of TES devices (TRANSAIR) were developed for indoor and outdoor usage. To increase the efficacy of TES, the frequency modulation according to normal distribution in the limits of the quasiresonance characteristics was put into operation. The blind and placebo-controlled (passive and active placebo) study was produced to estimate the TES effects on stress events and accompanied psycho physiological and autonomic disturbances of different intensities on volunteers and patients in the following groups: everyday stress and fatigue; stress in regular military service and in field conditions; stress in the relatives of those lost in mass disaster; posttraumatic stress (thermal burns); and affective disorders in a postabstinence period. Some subjective verbal and nonverbal tests and objective tests (including heart rate variability) were used for estimation of the initial level of psycho-physiological status, which changes after TES sessions. It was demonstrated that fatigue, stress, and other accompanied psycho physiological disturbances were significantly improved or abolished after 2-5 TES sessions. The TES effects were more pronounced in cases of heavier disturbances. In conclusion, activation of the brain endorphinergic structures by TES is an effective homeostatic method of FES that sufficiently improves quality of life. PMID- 11940026 TI - Simulation of the three-dimensional electrical field in the course of functional electrical stimulation. AB - Optimization of stimulation parameters as well as shape and positioning of electrodes are important questions in functional electrical stimulation (FES) of paraplegic patients. For that reason a MATLAB tool, called FES-FIELD, modeling the three-dimensional electric field in the human body, has been developed to calculate the electric field in a region of interest. The simulation tool provides a graphic user interface. In case of denervation of the lower extremities, an important target muscle is the m. quadriceps femoris. The electrical potential distribution along its fibers is representative for its functional activation. For this special application, the human thigh stimulated by skin electrodes was modeled. The simulation process was done in 5 steps: reading the geometric information of the thigh from 50 computed tomographic slices, segmentation in tissue types by pixel value and definition of each conductivity, selection of electrode geometry and positioning, calculating the electric field iteratively by solving the system of linear equations, and visualization of the solution by equipotential lines in either cross or length sections of the thigh. PMID- 11940027 TI - Clinical elements for the neuromuscular stimulation and functional electrical stimulation protocols in the practice of neurorehabilitation. AB - The physicians and their multidisciplinary teams involved in the clinical practice of neurological rehabilitation have more and more opportunities to apply neuromuscular stimulation (NMS) and functional electrical stimulation (FES) of peripheral nerves as a part of their daily practice. In this article, we outline clinical protocols of NMS and FES in the following clinical conditions of upper motor neuron dysfunction: to prevent consequences of disuse of the neuromuscular system of the upper motor neuron, to facilitate recovery processes of impaired upper motor neuron functions due to acute and/or subacute neurological conditions, to maintain or enhance the trophic state of the muscle, to modify altered control of muscle tone, to modify altered patterns of automatic and volitional functional movements, to enhance functional movement of the single joint muscle group within intact functional multijoint movement, and to modify altered neurocontrol of posture, locomotion, and skillful movements. We emphasize the importance of understanding the motor control alteration while developing clinical protocols and defining the goals. It is very important to be aware that similar clinical findings and due to the same cause can have different features of residual motor control, and therefore potentials for recovery or modification can be very different. PMID- 11940028 TI - Dermatome electrical stimulation as a therapeutic ambulatory aid for incomplete spinal cord injured patients. AB - Electrical stimulation of the L-3,4 dermatome during treadmill walking is proposed as a gait training modality in incomplete spinal cord injured patients. The dermatome stimulation proved to be efficient in diminishing the extensor tone occurring after loading of the paralyzed limb during the stance phase of walking and resulting in improved flexion of the leg during the swing phase. PMID- 11940029 TI - A pilot study to investigate the combined use of botulinum neurotoxin type a and functional electrical stimulation, with physiotherapy, in the treatment of spastic dropped foot in subacute stroke. AB - The objective was to inform sample size calculations for a full randomized controlled trial (RCT). The design included an RCT pilot trial with a 16 week study period, including a 4 week baseline phase. The subjects were adults within 1 year of first stroke, ambulant with a spastic dropped foot. Twenty-one participants were recruited from the stroke services of 4 centers. For intervention all participants received physiotherapy; the treatment group also received botulinum neurotoxin Type A (BoNTA) intramuscular injections to triceps surae (800 U Dysport) and functional electrical stimulation (FES) of the common peroneal nerve to assist walking. The main outcome measure was walking speed. The result was a significant upward trend in median walking speed for both the control (p = 0.02) and treatment groups (nonstimulated p = 0.004, stimulated p = 0.042). Trend lines were different in location (p = 0.04 and p = 0.009, respectively). In conclusion, there is evidence of an additional, beneficial effect of BoNTA and FES. Sufficient information has been gained on the variability of the primary outcome measure to inform sample size calculations for a full RCT to quantify the treatment effect with precision. PMID- 11940030 TI - An implantable two channel drop foot stimulator: initial clinical results. AB - This article reports preliminary results of pilot studies of a new implantable two channel drop foot stimulator. The stimulator consists of an externally worn transmitter inductively coupled to an implanted receiver unit located in the lower leg, lateral and distal to the knee. The receiver is connected to electrodes located under the epineurium of the deep and the superficial peroneal nerves. Stimulation is triggered by detection of heel lift and terminated at heel strike in a manner similar to surface mounted systems. The location of the electrodes allows for a degree of selectivity over the resultant moment about the ankle joint that is not possible with surface stimulation of the common peroneal nerve. The two subjects used the stimulator on a regular basis and showed increases in walking speed of between 10% and 44% when compared to their baseline measurements. Isometric tests have demonstrated that the stimulator allows selective and repeatable stimulation of ankle joint muscles. PMID- 11940031 TI - Restitution of reaching and grasping promoted by functional electrical therapy. AB - Functional electrical therapy (FET) is a new term describing a combination of functional electrical stimulation that generates life-like movement and intensive exercise in humans with central nervous system lesions. We hypothesized that FET can promote a significant recovery of functioning if applied in subacute stroke subjects. The study included 16 stroke subjects divided into a low functioning group (LFG) and a high functioning group (HFG) based on their ability to control wrist and fingers and randomly associated into FET and controls. The FET consisted of 30 min daily sessions during 3 weeks. The exercise comprised functional use of daily necessary activities (e.g., writing, using a telephone receiver, and drinking from a can). The outcome presented in this article is the upper-extremity function test performed before and after the therapy. The change in performance of the HFG group was significant. The number of successful repetitive movements in 2 min was doubled and 1.6 times increased for controls, and the time to perform the movement was decreased by 71% percent and by 36% in controls. In the LFG FET group, the difference in performance was the following. First, the number of tasks was increased from 0 to 6 (total of 11 tasks). Second, the averaged number of successful repetitive movements was increased from 0 to 3. The functional improvement in the FET LFG is probably not sufficient to make the more affected arm/hand effective for daily necessities; thus, the FET effects could deteriorate over a longer time. The subjects from the control LFG made only a marginal improvement. The follow-up for each subject will continue for 12 months after the beginning of the treatment. PMID- 11940032 TI - A stimulator for functional activation of denervated muscles. AB - In recent years various studies proved that electrical stimulation can improve contractile capability and restore muscle function in long-term denervated degenerated muscles. The low excitability of the muscle cells at the initial stage of training and surrounding connective tissue, acting as an electrical shunt, require special stimulation parameters. Until now, no appropriate devices (stimulators) are commercially available. Therefore, we were forced to design our own stimulators. The control unit of the stimulators is based on a microprocessor for maximum flexibility regarding the generation of the parameters such as pulse amplitudes, pulse width, frequency, stimulation times, ramps, and so on. In addition, the microprocessor design allows recording of compliance data such as stimulation date, time, duration, and used programs. The constant voltage output stage of the stimulator is able to generate biphasic charge balanced stimulation impulses with a pulse width of 1 to 300 ms, voltage amplitudes up to +/-80 V (160 VPP), and stimulation currents up to 250 mA. To prevent direct current due to inexact charge compensation, the electrode outputs are decoupled capacitively. Simultaneous 2 channel stimulation with independent intensity levels is possible. The stimulators are programmed using a notebook or a personal digital assistant via infrared serial interface. This concept guarantees the application of correct stimulation parameters because the patient has only access to parameters that are preprogrammed for him in the outpatient clinic. For the home based training, access is limited to variation of intensity within preprogrammed limits. For safety reasons, the portable unit is powered by an internal rechargeable battery. High efficiency switched voltage regulators are used to provide the different required voltage levels while ensuring an acceptable operating time of the stimulator. PMID- 11940033 TI - Twenty-eight years of clinical experience with implantable neuroprostheses for various applications. AB - Since 1973, the author has been implanting neural stimulators and later drug pumps to restore or improve motor function and modulate pain, spasticity, and seizures in patients with spinal cord and brain injury, cerebral palsy, stroke, and multiple sclerosis. During these 28 years, many physicians, biomedical engineers, and manufactures have realized worthwhile successes. Many lessons have been learned to improve operative techniques to ensure safety, low infection, and improved results for implant patients. The relationships between manufacturers and physicians have varied. Problems arise with patents, royalties, confidentiality, publishing, and liability insurance. There has been a need to patent ideas and intellectual properties; however, some of the patented concepts have been published previously but missed by the patent author and patent office. This has led to vigorous legal battles, consuming money with time delays, or resulting in surrendering worthwhile projects. There is a need for a responsible, independent appeals board to review these disputed patent claims. Then their findings should be admissible at the Patent Office and if necessary in court. PMID- 11940036 TI - Letter from the Editorial Team. PMID- 11940034 TI - A personal user's view of functional electrical stimulation cycling. AB - Two years of functional electrical stimulation cycling (FESC) as a researcher and subject have given me an insight into the direction that future FESC should take as well as providing me with significant health benefits and an enjoyable and functional ability to cycle. If FESC is to benefit spinal cord injured persons (SCIPs), researchers must turn their attention to making the activity convenient and enjoyable. What follows is a personal view and will be less scientifically rigorous than other presentations but hopefully still of value. It calls upon my experience as a general medical practitioner with a special interest in the value of exercise, a human powered vehicle enthusiast, an amateur FES researcher, but most importantly, an SCIP and FES cyclist. PMID- 11940037 TI - Towards improving the utility of fine-needle aspiration biopsy for the diagnosis of thyroid tumours. PMID- 11940038 TI - Oestrogen, bones and men: when testosterone just isn't enough. PMID- 11940039 TI - Height prognosis in girls with central precocious puberty treated with GnRH analogues. PMID- 11940040 TI - Factors predicting adult height in girls with idiopathic central precocious puberty: implications for treatment. AB - OBJECTIVES: To optimize the indications for treating girls with idiopathic central precocious puberty with GnRH analogues, since outcomes may vary. DESIGN: Comparison of adult heights with the data at initial evaluation and the target heights. PATIENTS: Group 1 patients (n = 43) were treated with GnRH analogue from 7.9 +/- 0.2 years to 10.8 +/- 0.1 years (bone age: 10.3 +/- 0.2 years to 12.2 +/- 0.1 years) and group 2 patients (n = 29) were monitored without treatment because their predicted adult heights were > 155 cm. The criteria for treatment were a predicted height < 155 cm and/or a LH/FSH peaks ratio of > 0.6. RESULTS: At initial evaluation, group 1 patients had greater breast development (P = 0.001) and bone age advances (2.0 +/- 0.2 years) than those of group 2 (1.3 +/- 0.2 years, P < 0.3), and higher plasma oestradiol concentrations (139 +/- 11 pmol/l vs. 62 +/- 7 pmol/l, P = 0.0001), LH peak (12.2 +/- 1.8 IU/l vs. 5.8 +/- 2.2 IU/l, P = 0.0001) and LH/FSH peaks ratio (1.3 +/- 0.2 vs. 0.5 +/- 0.1, P = 0.0001). The predicted height for group 1 at the onset of treatment (156 +/- 1.2 cm) was lower than the adult height (159.5 +/- 0.8 cm, P = 0.002), but the two were similar (164.1 +/- 1.2 cm vs. 162.7 +/- 0.9 cm) for group 2. In group 1, the difference between these heights (mean 3.4 cm) was positively correlated with the bone age advance (r = 0.51, P = 0.001), but not with chronological or bone ages, oestradiol, LH peak, LH/FSH peaks ratio before treatment or its duration. It was 5.3 +/- 1.2 cm in the 28 patients with a bone age advance of > 2 years and 0 +/- 1.3 cm in the other 15 (P < 0.02). It was 6.1 +/- 1.3 cm in the 24 patients with predicted height < 155 cm, and -0.1 +/- 1.1 cm in the other 18 (P = 0.002). The 72 patients included nine who attained an adult height over 5 cm shorter than the target height (seven treated and two untreated). The seven treated subjects included two who had retarded intrauterine growth. CONCLUSIONS: Treatment of girls with idiopathic central precocious puberty with GnRH analogues produced a mean height increase of 3.4 cm between the predicted and adult heights. The increase was greater for girls with a bone age advance of > 2 years and a predicted height < 155 cm. Adult height is spontaneously preserved in the slowly progressing form. The classical and slowly progressing forms can be distinguished by the degrees of breast development and bone age advance. PMID- 11940041 TI - Serum lipoprotein changes following IGF-I normalization using a growth hormone receptor antagonist in acromegaly. AB - OBJECTIVE: Active acromegaly is associated with altered lipid metabolism. The purpose of this study was to investigate the effect of serum IGF-I normalization on serum lipoproteins and insulin, in patients with acromegaly receiving the GH receptor antagonist pegvisomant. PATIENTS: Twenty patients (9 male, mean age 58.7 years, range 28-79) with active acromegaly (baseline serum IGF-I > 130% the age related upper limit of normal) received pegvisomant and achieved a normal serum IGF-I [585.2 +/- 54.3 (mean +/- SEM) to 169.2 +/- 13.9 ng/ml, P < 0.0001]. MEASUREMENTS: Total cholesterol (TC), high-density lipoprotein (HDL) cholesterol, triglyceride (TG), apolipoprotein B (apo B), apolipoprotein A1 (apo A1), lipoprotein a [Lp(a)] and insulin were measured in a single batch analysis on samples obtained at baseline and the first occasion of serum IGF-I normalization. Low-density lipoprotein (LDL) was calculated using the Friedewald formula. Paired analysis was performed using Student's paired t-test and the Wilcoxon signed rank test. RESULTS: Normalization of serum IGF-I resulted in an increase in TC (5.0 +/ 0.3 to 5.7 +/- 0.4 mmol/l, P = 0.0068), an increase in LDL (3.0 +/- 0.25 to 3.7 +/- 0.31 mmol/l, P = 0.0093) and an increase in apo B (110.6 +/- 7.76 to 127.1 +/ 8.86 mg/l, P = 0.014). TC and LDL increased in all but four patients. Despite a significant fall in fasting insulin levels (9.9 to 8.3 mU/l, range 8.85-19.8 to 6.33-11.6, P < 0.001) and insulin resistance (2.7 to 1.9, range 1.2-10.4 to 1 6.2, P < 0.001), mean serum TG and HDL levels were unaffected by IGF-I normalization. The protein component of HDL, apo A1, increased (153 +/- 4 to 166.4 +/- 5.43 mg/l, P = 0.026) and Lp(a) declined (median 342 to 235 mg/l, range 60-1013 to 74-671), P = 0.0035). Baseline serum TC and LDL were below the age- and sex-matched mean population value but after normalization of serum IGF-I the distribution of serum TC and LDL values was similar to that of the general population. CONCLUSIONS: Active acromegaly is associated with lowered mean serum TC and LDL. Successful management using pegvisomant increases lowered baseline serum TC and LDL levels, restoring the distribution of values to that of the general population, and improves insulin resistance. These findings are consistent with the reported lipoprotein changes following GH administration to normal and GH-deficient individuals. PMID- 11940042 TI - Postoperative persistent thyrotrophin releasing hormone-induced growth hormone release predicts recurrence in patients with acromegaly. AB - OBJECTIVE: To assess the predictive value of postoperative thyrotrophin releasing hormone (TRH)-induced GH responsiveness in relation to (late) postoperative outcome in patients in remission after surgery for acromegaly. PATIENTS: and methods One hundred and twenty-nine patients underwent surgery for acromegaly in our institution between 1977 and 1996. TRH tests and oral glucose tolerance tests (GTT) were performed and serum IGF-I concentrations were measured pre- and postoperatively and during follow-up. Criteria for postoperative remission were a mean serum GH concentration < 5 mU/l and/or serum GH after an oral glucose tolerance test < 1 mU/l immunofluorometric assay (IFMA) or < 2.5 mU/l (radioimmunosassay), together with a normal serum IGF-I concentration. RESULTS: Preoperatively, the TRH-induced GH response was highly variable, with gradual overlap between 'nonresponders' and 'responders'. Arbitrarily defined as a doubling of serum GH concentration, 45.6% of patients were 'responders' to TRH. GH response after TRH injection was significantly correlated to the TRH-induced prolactin response but not to preoperative GH concentration or adenoma size. After surgery, remission was achieved in 83 of the 129 patients. Postoperative remission was significantly correlated to mean preoperative serum GH concentration and preoperative glucose-suppressed serum GH but not to tumour class. Seventy-one patients with early postoperative remission were followed without adjuvant treatment for a mean of 9.4 +/- 0.7 years (range 0-23 years). Forty-one of these patients were TRH responsive as defined by at least doubling of the serum GH concentration preoperatively. Of the 71 patients, 12 developed recurrence of disease, as defined by insufficient GH suppression during oral GTT, and elevated IGF-I and mean serum GH concentration. Irrespective of the preoperative response to TRH, the initial postoperative TRH test was predictive of developing disease recurrence with a sensitivity of 75% and a specificity of 100% when an absolute GH increase of 3.75 mU/l was chosen to define paradoxical responsiveness. A stimulated GH 1.6 times basal was predictive of recurrence with a sensitivity of 83% and a specificity of 73%, and 2.1 times basal was predictive with a sensitivity of 75% and a specificity of 80%. None of the 32 patients with postoperative normalization of the preoperatively present TRH-induced GH response, defined as a postoperative GH increase < 3.75 mU/l, developed recurrence of disease, while all nine patients with a GH increase above this level developed recurrence of acromegaly. CONCLUSION: To our knowledge this is the first report which addresses the value of early postoperative TRH-induced GH responsiveness in predicting late surgical outcome using receiver-operating characteristic (ROC) curves to redefine a postoperative paradoxical response instead of arbitrarily chosen criteria. An absolute postoperative GH response of more than 3.75 mU/l was associated with recurrence in all nine patients, while all 32 patients with normalization of previously paradoxical response are still in remission. Our findings from this study demonstrate that the TRH test is a valuable tool in the early identification of patients at risk of developing postoperative recurrence of acromegaly. PMID- 11940043 TI - Relationships of urinary phyto-oestrogen excretion to BMD in postmenopausal women. AB - OBJECTIVE: Phyto-oestrogens are plant compounds with both oestrogenic and anti oestrogenic properties. However, it is not known whether natural phyto-oestrogens are beneficial or harmful in human osteoporosis. This study was performed to investigate the relationships between urinary phyto-oestrogens and bone mineral density (BMD) in Korean postmenopausal women. DESIGN: The subjects were classified into osteoporotic, osteopenic and normal groups according to their BMD as defined by WHO criteria. We compared the urinary phyto-oestrogens of each group and studied whether urinary phyto-oestrogens correlate with BMD. PATIENTS: The subjects were 75 Korean postmenopausal women with ages ranging from 52 to 65 years (mean 58 +/- 1.1 years). Mean number of years after menopause was 7.3 +/- 1.3. MEASUREMENTS: Twenty-four-hour urinary phyto-oestrogens were measured by gas chromatography-mass spectrometry (GCMS) and BMD by dual-energy X-ray absorptiometry (DXA, Lunar Expert-XL, Lunar Co., WI, USA). RESULTS: In Korean postmenopausal women, urinary enterolactone (1.46 +/- 1.11 micromol/day) was lower and daidzein (2.59 +/- 3.25 micromol/day) was higher than in western women, and both levels were comparable to those in Japanese women. Daily urinary excretion of genistein and apigenin were 1.09 +/- 0.912 and 0.48 +/- 0.40 micromol/day, respectively. In subjects with osteoporosis, urinary enterolactone was lower (P < 0.05) but apigenin was significantly higher (P < 0.05) than in the controls. BMD of L2-L4 correlated positively with urinary enterolactone (r = 0.388, P < 0.01), and BMD of the femoral neck and Ward's triangle correlated positively with urinary enterolactone (r = 0.271, P < 0.05 and r = 0.322, P < 0.05) but negatively with apigenin (r = -0.412, P < 0.01 and r = -0.395, P < 0.01). By multiple stepwise regression, the variables associated with spinal BMD were age, the amount of urinary apigenin and body mass index (BMI). The variables associated with femoral neck BMD were age and urinary apigenin. CONCLUSIONS: From these results, we conclude that urinary phyto-oestrogens, especially enterolactone and apigenin, are related to BMD in Korean postmenopausal women. Our results also suggest the possibility that phyto-oestrogens have differential effects on bone density. Further studies are needed to clarify the exact biological roles of phyto-oestrogenic components on bone metabolism. PMID- 11940044 TI - Growth hormone release after glucagon as a reliable test of growth hormone assessment in adults. AB - OBJECTIVE: To investigate the GH response to glucagon in adult patients with GH deficiency and in controls compared with the GH response to the insulin tolerance test (ITT) in patients with GH deficiency and to determine whether the use of glucagon results in a diagnostic utility test. PATIENTS AND DESIGN: Seventy-three patients with adult GH deficiency and organic hypothalamic-pituitary disease were recruited, along with 46 controls. The patients were divided into five groups according to the number of associated hormone deficiencies present. MEASUREMENTS: Hypopituitary subjects underwent assessment of GH secretory status by the ITT, the glucagon test and measurement of serum IGF-I concentration. Controls underwent the glucagon test. After the ITT, glucose and GH levels were measured at baseline, 30, 60 and 90 minutes, and after glucagon at baseline, 90, 120, 150, 180, 210 and 240 minutes. RESULTS: The highest GH value after the ITT in the patient group was 3 microg/l (0.76 +/- 0.82 microg/l), and after the glucagon test the highest GH peak value was 2.9 microg/l (0.64 +/- 0.79 microg/l). A correlation was found between the GH peak and the progressive number of hormone deficiencies. After the glucagon test, the GH peak obtained in the controls at 180 minutes was 9.8 +/- 4.6 microg/l and, on an individual basis, none of the 46 controls failed to achieve peak GH levels higher than 3 microg/l. In the controls, a negative correlation was observed between the GH response to glucagon and age (r = -0.389, P = 0.0075) and body mass index (r = -0.329, P = 0.0254). The accuracy of the glucagon test for differentiating patients from controls, estimated by receiver operating characteristics (ROC) curve methodology, showed that the cut-off of 3 microg/l for the GH peak provides 100% sensitivity and 100% specificity and is a reliable decision threshold. CONCLUSIONS: The glucagon GH test is reliable and provides a clear separation between GH-deficient and normal adults. A single glucagon test with a cut-off of 3 microg/l for the GH peak is diagnostic of GH deficiency in adults and could be considered and studied as an alternative to the ITT. PMID- 11940045 TI - Elevation of soluble E-selectin levels following gestational diabetes is restricted to women with persistent abnormalities of glucose regulation. AB - OBJECTIVE: Increased levels of the soluble adhesion molecule sE-selectin have been reported in normoglycaemic women with previous gestational diabetes but not in other groups at increased risk of future type 2 diabetes. To explore the basis for these discrepant findings, we studied the relationship between sE-selectin and glucose regulation in a large group of women with previous gestational diabetes. DESIGN: Comparison of sE-selectin levels between a study cohort ascertained on the basis of recent gestational diabetes and suitable control subjects. PATIENTS: One hundred and forty women with recent gestational diabetes (104 European, 20 South Asian and 16 Afro-Caribbean) and 125 normoglycaemic control women (90 European, 19 South Asian and 16 Afro-Caribbean). MEASUREMENTS: sE-selectin, fasting lipids, insulin and current glucose regulation status. RESULTS: There was no overall difference in sE-selectin levels between women with a history of gestational diabetes and control women among the 3 ethnic groups. European post-GDM women with abnormal glucose regulation postpartum (n = 30) had higher sE-selectin than control women (67 (54-91) ng/ml vs. 57 (43-75) ng/ml, P = 0.049). There was no difference in sE-selectin between normoglycaemic European women with a history of gestational diabetes (n = 74) and control women, even though the former displayed metabolic abnormalities predictive of diabetes. In those European post-GDM women with normal glucose regulation postpartum, sE selectin levels were negatively correlated with time since delivery (r = -0.25, P = 0.04), suggesting that the previously described elevation of sE-selectin following GDM pregnancies slowly resolves postpartum. There was no correlation between sE-selectin levels and features of insulin resistance. CONCLUSIONS: In contrast to previous findings, this larger study does not support a role for sE selectin as a marker, independent of prevailing glucose levels, of early metabolic abnormalities or future diabetes risk in women with previous gestational diabetes. PMID- 11940046 TI - Mechanisms of aneuploidy in thyroid cancer cell lines and tissues: evidence for mitotic checkpoint dysfunction without mutations in BUB1 and BUBR1. AB - OBJECTIVE: Thyroid follicular adenomas (FA) and carcinomas (FC) have a high prevalence of aneuploidy. We examined the contribution of mitotic checkpoint dysfunction and mutations of BUB1 or BUBR1, components of the spindle assembly checkpoint pathway, to chromosomal instability in thyroid cancer. DESIGN: The integrity of the mitotic checkpoint was studied in 8 aneuploid thyroid tumour cell lines. All cell lines as well as 9 FA, 9 FC, and 1 aneuploid papillary carcinoma were screened for mutations of BUB1 by SSCP and direct sequencing. Cell lines were also examined for mutations of BUBR1. RESULTS: Neither FRO, NPA nor WRO cells arrested in mitosis after treatment with nocodazole, whereas other aneuploid cell lines paused appropriately following microtubule disruption. One FC had a 2-bp somatic deletion (G2480/A2481) of BUB1 leading to a frameshift, and one FC had a silent polymorphism at nucleotide 1049 (TGT-TGC). There was a silent polymorphism of BUBR1 (G1271A) in one sample. CONCLUSION: Some, but not all thyroid cancer cell lines with aneuploidy have an abnormal mitotic checkpoint, indicating that chromosomal instability may arise through alternative cell cycle defects. Moreover, mutations of BUB1 or BUBR1 are infrequent in follicular neoplasms, and do not account for aneuploidy in thyroid cancer. PMID- 11940047 TI - Ventilation threshold as a measure of impaired physical performance in adults with growth hormone excess. AB - OBJECTIVE: Fatigue is a prominent symptom among patients with GH excess and acromegaly. Identifying the physiological basis of such complaints and obtaining objective measures to quantify their severity remains an ongoing challenge. We investigated whether submaximal measures of aerobic performance can be used to assess GH excess-associated fatigue objectively. DESIGN AND PATIENTS: To investigate this possibility we examined the relation between physical function and physical capacity in 12 patients with active acromegaly and persistent fatigue before and after 3 and 6 months of treatment with the long-acting somatostatin analogue octreotide (LAR(R)). MEASUREMENTS: Heart rate (HR) and rating of perceived exertion (RPE using Borg's 10-point scale) were measured during a 160-metre self-paced walk test (SPW). Maximum oxygen uptake (VO2max) and ventilation threshold (VeT: a measure of work rate when breathlessness develops) were measured during a progressive treadmill test to fatigue or symptom-limited maximum. The Profile Of Mood States questionnaire (POMS) was used to quantify subjective feelings of fatigue and vigour. Morning fasting levels of GH and IGF-I were measured using immunoassay of serum samples. RESULTS: SPW speed at a fast pace of 1.69 +/- 0.18 m/s was achieved with higher than normal HR (112 +/- 15/min; normal = 102) and RPE (2.4 +/- 1.2). Similar to GH-deficient adults, VO2max (22.6 +/- 6.4 ml.kg-1.min-1; normal approximately 30 ml.kg-1.min-1) and VeT (13.1 +/- 2.9 ml.kg-1.min-1; predicted normal approximately 16 ml.kg-1(min-1) were low. However, VeT occurred at a normal fraction of VO2max (VeT/VO2max = 0.58). VeT was significantly increased and plasma IGF-I levels reduced following 3 and 6 months of octreotide LAR(R) treatment. Reduction in circulating IGF-I levels was correlated with improvement in reported vigour (r = 0.85) and VeT (r = 0.65) (P < 0.05). CONCLUSIONS: Our findings demonstrate impairment in physical function and physical capacity consistent with the perception of increased fatigue among acromegalic patients. These objective measures of compromised physical function are similar to the changes that we have reported previously in adults with GH deficiency. Taken together, these data suggest that a narrow window for GH/IGF-I levels is required to maintain optimal physical function. PMID- 11940048 TI - Poor uterine development in Turner syndrome with oral oestrogen therapy. AB - OBJECTIVE: To evaluate uterine development in Turner syndrome (TS) patients in relation to treatment with oral ethinyl oestradiol (E2 ) for pubertal induction. DESIGN AND PATIENTS: Pelvic ultrasound data for 96 TS patients scanned since 1989 were analysed. Patients were classified into three groups: (1) untreated (n = 48); (2) complete spontaneous puberty (n = 10); and (3) treated with ethinyl oestradiol (n = 38). Uterine development was described in the three groups and compared with the normal data. MEASUREMENTS: Uterine length, fundal-cervical ratio (FCR) and shape were recorded, and presence or absence of ovaries noted. In the treated group, cross-sectional and longitudinal data were combined to compare uterine development with Tanner breast stage. RESULTS: In untreated girls up to age 10 years there was a variable distribution of uterine length and FCR about the mean. Thereafter, the uterus failed to grow and mature normally. Girls with complete spontaneous puberty had morphologically normal ovaries and uteri, but of 7 girls who attained menarche, 3 subsequently developed secondary oligomenorrhoea or amenorrhoea. In the treated group, in general, breast development and uterine length progressed with increasing E2 dose. However, only 50% of girls with complete secondary sexual development had a mature heart-shaped uterine configuration. CONCLUSIONS: Our current E2 treatment regimen for TS girls gives rise to satisfactory pubertal induction and maintenance, but failed to induce a fully mature uterus in half the cohort. In view of the high risk of miscarriage in TS in both spontaneous and assisted pregnancies, the effect of more physiological methods of E2 replacement on uterine development should be investigated. PMID- 11940050 TI - Long-term follow-up of patients with bone metastases from differentiated thyroid carcinoma -- surgery or conventional therapy? AB - OBJECTIVE: Surgery of bone metastases from differentiated thyroid carcinoma seems indicated in individual patients. This study was performed (1) to analyse retrospectively patients with bone metastases from differentiated thyroid carcinoma and (2) to evaluate the impact of surgery of bone metastases on survival. PATIENTS AND DESIGN: We analysed 41 consecutive patients with bone metastases from differentiated thyroid carcinoma who had undergone thyroid surgery at Vienna University Hospital since 1966. The median follow-up time was 12 years. There were 24 females and 17 males with a mean age of 60 +/- 12 years. Primary tumour histology was follicular in 35 and papillary in six patients. Radioiodine treatment was performed in 32 with a mean administered activity of 27 +/- 24 GBq 131I. Metastases restricted to the skeleton were found in 22 whereas in 19 individuals additional extraskeletal distant metastases were seen. Twenty seven patients had multiple bone metastases. In 21 individuals, up to five bone metastases were surgically removed with the intention of cure. RESULTS: Univariate analysis identified total thyroidectomy (P = 0.003), lymph node surgery (P = 0.001), radioiodine therapy (P = 0.036), and the absence of extraskeletal distant metastases (P = 0.017) as significant predictors of survival. Multivariate analysis failed to identify significant prognostic factors. In the subgroup of patients with distant metastases limited to the bones, univariate analysis identified, in addition to thyroid and lymph node surgery, the surgical extirpation of the bone metastases as a significant prognostic factor associated with improved survival (P = 0.025). CONCLUSIONS: These findings indicate that in patients without additional extraskeletal distant metastases, the radical surgical extirpation of bone metastases from differentiated thyroid carcinoma might be associated with improved survival. PMID- 11940049 TI - Mutations linked to familial hypokalaemic periodic paralysis in the calcium channel alpha1 subunit gene (Cav1.1) are not associated with thyrotoxic hypokalaemic periodic paralysis. AB - OBJECTIVE: To investigate whether patients with thyrotoxic hypokalaemic periodic paralysis (THPP) have the same molecular defect in the calcium channel gene described in familial hypokalaemic periodic paralysis (FHPP), as the symptoms of both diseases are comparable, we analysed, in patients with THPP, the presence of mutations R528H, R1239H and R1239G on the S4 voltage-sensing transmembrane segment of the alpha1 subunit of the calcium channel gene (Cav1.1). DESIGN AND PATIENTS: Genomic DNA was extracted from peripheral blood from 14 patients with THPP, 13 sporadic cases and one with a family history. An FHPP family was selected as a positive control. The exons bearing the described mutations were amplified by PCR, screened by single-strand conformation polymorphism (SSCP), and further sequenced. MEASUREMENTS: THPP was diagnosed both clinically and through laboratory tests, all patients having elevated levels of thyroid hormones (T4, T3 or free T4), suppressed TSH and plasma potassium below 3 small middle dot5 mmol/l. RESULTS: No evidence of the described mutations was found in patients with THPP. Furthermore, we did not detect any mutations in any of the four full S4 voltage-sensing transmembrane segments of Cav1 small middle dot1 (DIS4, DIIS4, DIIIS4 and DIVS4) by direct sequencing. However, close to the R528H mutation, we identified two single nucleotide polymorphisms at nucleotides 1551 and 1564 in both familial and sporadic cases with THPP. In addition, we were able to detect the R528H mutation in the DIIS4 transmembrane segment in all members of the FHPP family. CONCLUSION: Mutations linked to familial hypokalaemic periodic paralysis in the calcium channel alpha1 subunit gene (Cav1.1) are not associated with thyrotoxic hypokalaemic periodic paralysis. However, polymorphisms in nucleotides 1551 and 1564 in the exon 11 were found in patients with familial hypokalaemic periodic paralysis and thyrotoxic hypokalaemic periodic paralysis in higher frequency than in controls. The polymorphisms identified within the Cav1.1 gene are associated with thyrotoxic hypokalaemic periodic paralysis and represent a novel finding. PMID- 11940051 TI - The relationship between insulin secretion, the insulin-like growth factor axis and growth in children with cystic fibrosis. AB - OBJECTIVE: Cystic fibrosis-related diabetes mellitus (CFRD) is an increasingly common complication of cystic fibrosis. CFRD is preceded by a progressive decline in insulin secretion but there is no accepted definition of the prediabetic state in CFRD. This prediabetic state appears to have adverse effects on clinical status, nutrition and lung function, but there is no direct evidence that the impaired glucose homeostasis is the cause of these deteriorations. This study examined the prevalence of glucose intolerance and impaired insulin secretion in a population of children with CF without CFRD. Severe CF lung disease is often associated with poor weight gain and slower growth but the mechanism for this is still unclear. The relationships between the current state of glucose homeostasis, insulin secretion and the insulin-like growth factor axis, height velocity, nutrition status and lung function were therefore studied. DESIGN AND PATIENTS: Eighteen children with cystic fibrosis aged 9.5-15 years had oral glucose tolerance tests and 14 of these also had intravenous glucose tolerance tests (four refused). Blood samples were collected for insulin, C-peptide, glucose, HbA1c, insulin-like growth factor (IGF)-I, IGF-II, IGF-binding protein (IGFBP)-1 and IGFBP-3. Data on height, weight, puberty status, clinical score (Shwachman score) and lung function were recorded. Height velocity, height and weight standard deviation scores (SDS) were calculated using WHO/CDC data. RESULTS: The mean height SDS (-0.52 +/- 0.17) was less than the normal population (P = 0.007) and the mean height velocity was 4.6 +/- 0.5 cm/year, 39% with a height velocity less than the third percentile for age. The weight SDS and body mass index (BMI) were similar to the normal population. Four children had impaired glucose tolerance. The first-phase insulin response (FPIR) was below the first percentile of normal population values in nine (65%). Impaired FPIR or impaired glucose tolerance did not correlate with the Shwachman score, nutritional status or pulmonary function. There was a significant positive correlation between insulin secretion (area under the curve) and height velocity (P = 0.001) and serum IGFBP-3 levels (P = 0.001). CONCLUSIONS: Impaired glucose tolerance was present in 20% of children with cystic fibrosis. Impaired insulin secretion was common (65%) even in children with normal glucose tolerance. The mean height SDS for the group was low and the height velocity was abnormally slow in 39%, yet nutritional status as measured by BMI was appropriate for age. Relative insulin deficiency rather than nutritional deprivation or poor clinical status thus appears to be implicated in the poor linear growth of these children with relatively stable lung disease. This was a small study and firm conclusions on this chronic suppurative disease as to the cause of poor growth are not possible. The causes of poor growth are likely to be complex; nevertheless, the apparent decrease in insulin secretion combined with the expected increased demands on insulin production during pubertal growth raises the question as to whether insulin therapy should be considered in children with cystic fibrosis before the onset of cystic fibrosis-related diabetes mellitus. PMID- 11940052 TI - Indirect evidence for decreased hypothalamic somatostatinergic tone in anorexia nervosa. AB - OBJECTIVE: In animals, somatostatin (SRIH) and growth hormone (GH)-releasing hormone (GHRH) increase feeding via a common neural mechanism. Furthermore, SRIH counteracts the suppressive action of corticotrophin-releasing hormone (CRH) on food intake. Hypothetically, SRIH could be involved in the central feeding mechanism in anorexia nervosa (AN). Peripheral administration of pyridostigmine (PD) minimizes the release of hypothalamic SRIH. DESIGN: To study the influence of hypothalamic somatostatinergic inhibition on the exaggerated somatotroph responsiveness to GHRH in patients with severe AN, two GHRH stimulation tests were performed in random order following pretreatment with placebo or PD 2 mg/kg body weight in 13 patients and in 10 age-matched healthy controls. The test procedure was repeated in the patients after weight gain. RESULTS: In controls, PD potentiated the GHRH-stimulated GH rise but this effect was absent in AN patients. The relative potentiating effect of PD was inversely correlated to cortisol excretion levels and positively correlated to leptin serum levels. After weight gain the relative PD effect increased twofold. CONCLUSION: The pyridostigmine-GHRH responsive pattern points indirectly to greater SRIH withdrawal and greater GHRH release in anorexia nervosa. Moreover, hypothalamic SRIH activity seems to be inversely related to cortisol levels, indirectly supporting the hypothesis that SRIH and CRH neuronal activity are inversely related in anorexia nervosa. Leptin, which is believed to act on hypothalamic feeding mechanisms, seems to be positively related to SRIH activity. Finally, the present data demonstrate that the potentiating effect of pyridostigmine in anorexia nervosa is related to body mass index and increases upon weight gain, suggesting that the low somatostatinergic tone is not primary but is related to the weight loss. PMID- 11940053 TI - Galanin in human pituitary adenomas: frequency and clinical significance. AB - OBJECTIVES: Galanin (GAL) is a neuropeptide widely expressed in the central and peripheral nervous system and in neuroendocrine tissue, including the adenohypophysis where, in humans, it is expressed in corticotrophs and in ACTH producing adenomas. Previous analyses of human tissue have used antiserum against porcine GAL for detection of GAL immunoreactivity (GAL-IR) and no pathophysiological correlates have been reported. Given significant differences between the sequence of porcine and human GAL peptides, the aim of this study was to use antiserum raised against synthetic human GAL to investigate GAL-IR in non tumorous pituitaries and in pituitary adenomas, and to correlate GAL-IR with the clinical and hormonal characteristics of patients with Cushing's disease. PATIENTS: Six nontumorous pituitaries were obtained from autopsy and 151 pituitary adenomas, comprising 62 functioning (16 corticotroph, 26 somatotroph, 19 lactotroph and one thyrotroph) and 89 nonfunctioning adenomas, were obtained by surgery. RESULTS: All non tumorous pituitary glands showed GAL-IR in corticotrophs, in basophil cells within the neurohypophysis and in nerve fibres of the neurohypophysis. GAL-IR was found in a subset (10 of 16) of patients with ACTH-secreting tumours causing Cushing's syndrome. GAL-IR was rarely expressed in somatotroph adenomas and prolactinomas, but was expressed in approximately one third of nonfunctioning tumours. GAL-IR was found in almost 90% of nonfunctioning tumours that were positive for ACTH. There were no significant differences in sex ratio, age at presentation or 24-h urinary free cortisol secretion in the subset of patients with Cushing's disease positive (n = 10) or negative (n = 6) for GAL IR. However, Cushing's patients positive for GAL-IR tended to have smaller tumours and achieved a higher cure rate than those without (100 vs. 50%, P = 0.017). CONCLUSIONS: Galanin is present in normal and tumorous human pituitaries. In addition, GAL colocalizes exclusively in corticotrophs of normal pituitaries and is coexpressed almost exclusively in corticotrophs from functioning and nonfunctioning tumours. The finding that corticotroph adenomas can function irrespective of the presence of GAL suggests that GAL may not play a pathophysiological role in Cushing's disease. However, the better surgical outcome observed in patients with Cushing's disease who had tumours positive for GAL-IR suggests that the expression of GAL confers a less aggressive tumour phenotype. PMID- 11940054 TI - Demonstration of anti-TSH antibody in TSH binding inhibitory immunoglobulin positive sera of patients with Graves' disease. AB - OBJECTIVES: The presence of anti-TSH antibodies in Graves' patients with unusually low TSH binding inhibitory immunoglobulin (TBII) has been reported. Recently, we found the first case of an anti-TSH antibody in TBII-positive sera of patients with Graves' disease. The prevalence and immunological specificity of this anti-TSH antibody were examined. DESIGN: The presence of 125I-bovine(b) TSH binding antibody in TBII positive serum was examined by prolonged incubation of more than 1 day because only weak binding occurred after 1 h incubation at 37 degrees C. The clinical course of these patients and binding characteristics of anti-bTSH antibody were examined. RESULTS: The corrected method-TBII activity (%)[1 - (a - b)/(c - d)] x 100 and the standard method-TBII activity (%) [1 - (a d)/(c - d)] x 100 [a, 125I-bTSH binding with TSH receptor (R) in the presence of test serum; b, 125I-bTSH binding with test serum; c, 125I-bTSH binding with TSH R in the presence of normal serum; d, 125I-bTSH binding with normal serum] were calculated. The corrected method-TBII activity was always higher than the standard method-TBII activity in anti-bTSH antibody-positive serum. Anti-bTSH antibody-positive cases in TBII-positive Graves' disease were found in approximately 1% of Graves' patients. Anti-bTSH antibodies were confirmed as IgG from the increase of precipitated radioactivity by adding rabbit antihuman IgG antibody after the incubation of 125I-bTSH with test serum. These antibodies bind with not only bTSH, bTSH(alpha) and bLH, but also porcine (p)TSH, pTSH(alpha) and pFSH. However, these antibodies did not bind with human TSH. Binding of 125I-bTSH with patient's serum was neither inhibited by other Graves' thyroid stimulating antibody (TSAb), nor thyroid blocking antibody (TBAb) in primary hypothyroidism. CONCLUSIONS: The presence of anti-bTSH antibody in TBII-positive serum of high titre means that TBII-positive sera cannot rule out the absence of anti-bTSH. Thus, determination of 125I-bTSH binding with test serum in TSH receptor assays is necessary to determine the precise TBII activity and to detect anti-bTSH antibody. PMID- 11940055 TI - Severe hypomagnesaemia-induced hypocalcaemia in a patient with Gitelman's syndrome. AB - Gitelman's syndrome (GS) is characterized by hyperreninaemic hyperaldosteronism, hypokalaemia, metabolic alkalosis, hypomagnesaemia and hypocalciuria and is due to a defect of the Na-Cl cotransporter at the distal tubule, which may appear in a sporadic or in a familial form. It is an autosomal recessive disorder associated with normal or reduced blood pressure. We report a case of severe hypomagnesaemia-induced hypocalcaemia in a 39-year-old Caucasian woman with GS. The patient had impaired parathormone (PTH) responsiveness to peripheral stimuli, as proved by the marked PTH increase and normalization of plasma calcium levels after acute and chronic administration of magnesium salts. Secondary normotensive hyperreninaemic hyperaldosteronism with hypokalaemia and metabolic alkalosis was also present. Normal plasma renin activity (PRA) and aldosterone levels were restored by administration of an inhibitor of prostaglandin synthesis. The electrolyte imbalance was successfully corrected with chronic treatment with magnesium and potassium salts. Genetic analysis identified a compound heterozygous mutation in the Na-Cl cotransporter gene (NCCT), confirming the diagnosis of GS. The striking feature of this case of GS was impaired PTH responsiveness to peripheral stimuli determined by hypomagnesaemia and the resulting severe hypocalcaemia, which had not previously been described in this syndrome. PMID- 11940056 TI - PTHrP during petrosal sinus sampling. PMID- 11940057 TI - Studies of cord blood mononuclear cell responses and allergy: still in their infancy? PMID- 11940058 TI - Relevance of early or current pet ownership to the prevalence of allergic disease. PMID- 11940059 TI - The role of COX-1 and COX-2 in asthma pathogenesis and its significance in the use of selective inhibitors. PMID- 11940060 TI - Regulation of the adaptive immune response to inhaled allergens. PMID- 11940061 TI - Glucocorticoids and immunoglobulin E: no new in vivo paradigms. PMID- 11940062 TI - New treatments for atopic dermatitis. PMID- 11940063 TI - Cord blood mononuclear cell cytokine responses in relation to maternal house dust mite allergen exposure. AB - BACKGROUND: Cord blood mononuclear cells have demonstrated specific immune responses to environmental allergens. OBJECTIVE: To establish whether the nature of this response is related to the level of maternal antenatal exposure to house dust mite (HDM) allergen and, hence, whether antenatal allergen avoidance may have a role in the prevention of allergic sensitization in children. METHODS: Children with a family history of asthma were recruited antenatally as subjects in a randomised controlled trial: the Childhood Asthma Prevention Study. HDM allergen (Der p 1) concentrations were measured in dust collected from the maternal bed at 36 weeks gestation. Cord blood mononuclear cells were stimulated in culture, separately, with phytohaemaglutinin (PHA) and HDM extract. Cytokine IL-4, IL-5, IL-10 and IFN-gamma concentrations in supernatant were measured by ELISA. mRNA signals for these cytokines were measured using RT-PCR. RESULTS: The median concentration of HDM allergen was 18.4 microg/g (interquartile range 7.3 35.3 microg/g). Median concentrations of IL-4, IL-5, IL-10 and IFN-gamma, after PHA stimulation were 4, 19, 401 and 1781 pg/mL, respectively. After HDM allergen stimulation the median concentrations were 0, 0, 20 and 14 pg/mL, respectively. The distribution of mRNA cytokine signals was similar. Neither cytokine protein concentrations nor cytokine mRNA signal levels were correlated with the concentration of HDM allergen in the mothers' beds at 36 weeks gestation. CONCLUSION: These findings do not support the view that the prevention of allergic disease in children requires the institution of HDM avoidance interventions during pregnancy. PMID- 11940064 TI - Early, current and past pet ownership: associations with sensitization, bronchial responsiveness and allergic symptoms in school children. AB - BACKGROUND: Studies have suggested that early contact with pets may prevent the development of allergy and asthma. OBJECTIVE: To study the association between early, current and past pet ownership and sensitization, bronchial responsiveness and allergic symptoms in school children. METHODS: A population of almost 3000 primary school children was investigated using protocols of the International Study on Asthma and Allergies in Childhood (ISAAC). Allergic symptoms were measured using the parent-completed ISAAC questionnaire. Sensitization to common allergens was measured using skin prick tests (SPT)s and/or serum immunoglobulin (Ig)E determinations. Bronchial responsiveness was tested using a hypertonic saline challenge. Pet ownership was investigated by questionnaire. Current, past and early exposure to pets was documented separately for cats, dogs, rodents and birds. The data on current, past and early pet exposure were then related to allergic symptoms, sensitization and bronchial responsiveness. RESULTS: Among children currently exposed to pets, there was significantly less sensitization to cat (odds ratio (OR) = 0.69) and dog (OR = 0.63) allergens, indoor allergens in general (OR = 0.64), and outdoor allergens (OR = 0.60) compared to children who never had pets in the home. There was also less hayfever (OR = 0.66) and rhinitis (OR = 0.76). In contrast, wheeze, asthma and bronchial responsiveness were not associated with current pet ownership. Odds ratios associated with past pet ownership were generally above unity, and significant for asthma in the adjusted analysis (OR = 1.85), suggesting selective avoidance in families with sensitized and/or symptomatic children. Pet ownership in the first two years of life only showed an inverse association with sensitization to pollen: OR = 0.71 for having had furry or feathery pets in general in the first two years of life, and OR = 0.73 for having had cats and/or dogs in the first two years of life, compared to not having had pets in the first two years of life. CONCLUSION: These results suggest that the inverse association between current pet ownership and sensitization and hayfever symptoms was partly due to the removal of pets in families with sensitized and/or symptomatic children. Pet ownership in the first two years of life only seemed to offer some protection against sensitization to pollen. PMID- 11940065 TI - Atopic sensitization among children in an arctic environment. AB - BACKGROUND: Asthma has been reported to be rare among Inuits, but so far total and specific IgE levels have never been determined in arctic populations. OBJECTIVE: To determine the prevalence of atopy in children living in an arctic environment, and to examine whether atopy and total IgE levels were associated with parental place of birth, as a measure of ethnicity, and travel history. MATERIAL AND METHODS: All schoolchildren in Sisimiut, a community on the West coast of Greenland, were screened for atopy. Blood samples were analysed for total IgE and for specific IgE against inhalant and food allergens. Information on place of birth of children and their parents was obtained from national registries. Information on travel history was obtained from self-administered questionnaires. RESULTS: A total of 1031 schoolchildren aged 5 to 18 years had a blood sample drawn (85% of available children for the study). Of these, 151 (14.6%) children were sensitized to at least one inhalant allergen and 42 (4.1%) to at least one food allergen. Sensitization to grass was most common, whereas sensitization to mugwort, birch, animal-dander and house-dust mite was infrequent. Children whose parents were both born abroad had a higher risk of sensitization to inhalant allergens compared with children born of Greenlandic parents (OR = 8.6, 95% CI 2.8-27.1). Furthermore, children who had been abroad had a higher risk of sensitization towards pollen (OR = 1.6, 95% CI 1.0-2.5) and animal-dander (OR = 2.1, 95% CI 1.0-4.6) after adjustment for confounders. Both atopic and non-atopic children demonstrated high levels of total IgE (medians of 251 and 58 kU/L). CONCLUSIONS: Compared with European findings Greenlandic children have high levels of total IgE but a low prevalence of allergic sensitization towards inhalant allergens. This may be due to a low genetic susceptibility to atopy and less allergen exposure, as well as to living conditions in an arctic environment. PMID- 11940066 TI - Increase of allergen-specific immunoglobulin E antibodies from 1973 to 1994 in a Finnish population and a possible relationship to Helicobacter pylori infections. AB - BACKGROUND: The prevalence of atopic diseases--hayfever, asthma and eczema--has increased over the past decades. The increase may be associated with decreased rates of infections such as measles, hepatitis A, tuberculosis, toxoplasmosis, and, as recently suggested, Helicobacter pylori gastritis. OBJECTIVE: Since the increase of atopy has been mainly based on clinical studies, we wanted to study the prevalence of allergen-specific Immunoglobulin (Ig)E antibodies in two cross sectional, adult population-based serum samples two decades apart. Since the sera had been tested for H. pylori antibodies, we also had a chance to look for a possible relationship between these two findings. METHODS: We determined the prevalence rate of allergen-specific serum IgE antibodies against birch and timothy pollen, and cat and dog epithelium allergens by the radioallergosorbent test in a 15-54-years-old Finnish population using 326 sera collected in 1973 and 319 sera collected in 1994 from randomly selected subjects. RESULTS: From 1973 to 1994 allergen-specific IgE prevalence rates and IgE antibody levels rose. In 1994, the prevalence rate of positive findings in 15-24-year-old population had increased from 11 to 38% (3.5-fold increase, P = 0.0001, OR 5.12, CI 95% 2.32 11.3). In older 10-year age groups similar trends did not reach significance, but the overall change was significant with all three cut-off levels of allergen specific IgE analysed. The percentage of IgE-positive persons rose mainly in the subgroup with no H. pylori antibodies. In 1994 21% of the H. pylori-negative subjects had IgE antibodies compared with 5% of the H. pylori-positive subjects (in 1973 11% in both subgroups). CONCLUSIONS: IgE-based evidence for an increase in IgE-mediated allergy was uncovered. The increase occurred mainly in the subgroup with no antibodies to H. pylori, which support the hypothesis that H. pylori could be one of the microbes counteracting atopy. PMID- 11940067 TI - The protective role of country living on skin prick tests, immunoglobulin E and asthma in adults from the Epidemiological study on the Genetics and Environment of Asthma, bronchial hyper-responsiveness and atopy. AB - BACKGROUND: Farming environment and traditional lifestyle seem to protect from childhood allergy. OBJECTIVE: The aim is to analyse the relationships of living in the country to asthma, positive skin prick tests and IgE among adults considering various windows of exposure over the life-span. METHODS: The study concerns 805 adults drawn from the Epidemiological study on the Genetics and Environment of Asthma, bronchial hyper-responsiveness and atopy (EGEA) (asthmatic cases, non-asthmatic controls, and parents of cases with and without asthma). Ever living in the country concerned 55% of the subjects. Early (beginning < 1 years), childhood (beginning < or = 16 years), prolonged (duration > or = 10 years) and current life in the country were studied. RESULTS: The results based on the case control and family components of the study show that IgE levels were significantly lower in those who ever lived in the country and in particular in those who lived for > or = 10 years. Positive skin prick tests (SPT) were significantly less prevalent in those who ever lived in the country and in particular in those with childhood (< or = 16 years) exposure. These associations remained independent of age, sex, smoking or asthma with IgE levels of 64 vs. 88 IU/mL; P = 0.004 for those ever living in the country vs. others and odds ratio for SPT positivity of 0.72 (95% CI [0.53-0.98]). In the more specific group with traditional mode of heating in childhood (use of wood) associations were stronger. The association with asthma, studied in parents of asthmatic probands showed that fathers, but not mothers, of asthmatics were significantly less often asthmatic themselves in relation to country living. CONCLUSION: Country life protects from asthma and adulthood allergy. The protective effect is not restricted to exposure in early childhood. PMID- 11940068 TI - Lack of association between a polymorphism in the interleukin-13 gene and total serum immunoglobulin E level among nuclear families in Costa Rica. AB - BACKGROUND: IL-13 has been implicated in the pathogenesis of asthma and in the regulation of IgE synthesis in humans. Single nucleotide polymorphisms (SNPs) in the IL-13 gene have been associated with asthma and total serum IgE level in Caucasian populations. OBJECTIVE: To test for genetic association between an SNP in exon 4 of the IL-13 gene (IL-13 + 2044 or Arg130Gln) and total serum IgE level and asthma-related phenotypes in a population with high prevalence of asthma living in Costa Rica. METHODS: Family-based association study. RESULTS: Among 83 Costa Rican school children with asthma and their parents (249 individuals), there was no evidence of linkage disequilibrium between the IL-13 + 2044 SNP and any of the outcomes of interest (total serum IgE level on a logarithmic scale, number of positive skin tests to aeroallergens, and asthma). These results were not significantly changed after adjustment for age and gender. CONCLUSIONS: No significant evidence of linkage disequilibrium between an SNP in exon 4 of the IL 13 gene and total serum IgE level, sensitization to allergens or asthma was found in a family-based association study in Costa Rica. PMID- 11940069 TI - Effect of ozone exposure on allergic sensitization and airway inflammation induced by dendritic cells. AB - BACKGROUND: Epidemiological studies suggest that ozone exposure is related to increased asthma symptoms. Dendritic cells (DCs) are the principal antigen presenting cells in the airways. OBJECTIVE: We have examined whether ambient doses of ozone (100 ppb for 2 h) enhance allergic sensitization and/or airway inflammation in a mouse model. METHODS: C57BL/6 mice were sensitized to inhaled ovalbumin (OVA) by intratracheal instillation of OVA-pulsed DCs on day 0. Daily exposure to OVA aerosol on days 14-20 resulted in an eosinophilic airway inflammation, as reflected in bronchoalveolar lavage fluid and lung histology. In a first experiment, mice were exposed to ozone or room air immediately prior to and following sensitization. Subsequently, we tested the effect of ozone exposure during antigen challenge in DC-sensitized mice. RESULTS: Exposure to ozone during sensitization did not influence airway inflammation after subsequent allergen challenge. In contrast, in sensitized mice, challenge with OVA together with ozone (days 14-20) resulted in enhanced airway eosinophilia and lymphocytosis, as compared with mice exposed to OVA and room air (1.91 x 106 +/- 0.46 x 106 vs. 0.16 x 106 +/- 0.06 x 106 eosinophils/mL lavage fluid; P = 0.015; 0.49 x 106 +/- 0.11 x 106 vs. 0.08 x 106 +/- 0.03 x 106 lymphocytes/mL lavage fluid; P = 0.004). Ozone exposure without subsequent OVA exposure did not cause airway inflammation. CONCLUSION: Ozone exposure does not increase allergic sensitization but enhances antigen-induced airway inflammation in mice that are sensitized via the airways. PMID- 11940070 TI - Safety of rofecoxib in subjects with a history of adverse cutaneous reactions to aspirin and/or non-steroidal anti-inflammatory drugs. AB - BACKGROUND: Adverse reactions to non-steroidal anti-inflammatory drugs (NSAID)s are frequent, and the need to identify a safe alternative drug is a common problem in clinical practice. OBJECTIVE: To assess the tolerability of rofecoxib, a drug that specifically inhibits COX-2, in a group of NSAID-sensitive patients. METHODS: One-hundred and four subjects (29 males and 75 females, mean age 35.6 +/ 14.1) were enrolled. All subjects had experienced one or more episode characterized by cutaneous symptoms (erythema, and/or urticaria angioedema) following the assumption of NSAIDs; 92 subjects experienced reactions to only one NSAID (single intolerance: SI) and 12 subjects had reactions to multiple NSAIDs (multiple intolerance: MI). Rofecoxib was challenged at the following dosages: 1/4 of 25 mg (6.25 mg), 1/4 of 25 mg, and 1/2 of 25 mg (12.5 mg), at intervals of 1 h if no symptoms had developed with the previous administration, in order to reach a cumulative dose of 25 mg. All subjects underwent two double-blind, placebo-controlled challenges in two consecutive days. RESULTS: No reactions against placebo were observed. Similarly, no reactions were observed in all subjects both after the first and after the second challenge to rofecoxib. CONCLUSIONS: The present study demonstrated that rofecoxib does not have cross reactivity to NSAIDs. Rofecoxib is a safe alternative in subjects with previous adverse cutaneous reaction to NSAIDs. PMID- 11940071 TI - In vivo kinetics of the immunoglobulin E response to allergen: bystander effect of coimmunization and relationship with anaphylaxis. AB - BACKGROUND: Murine models of hypersensitivity to allergens are useful tools for the evaluation of preclinical strategies to down-regulate the IgE response. OBJECTIVE: To monitor the long-term kinetics of T and B cell responses to allergen as a function of allergen dosage and to investigate the effect of parallel immunization with a second antigen; to correlate B cell response with anaphylaxis. METHODS: CBA/J mice were sensitized every other week by subcutaneous injections of phospholipase A2 (PLA2) and/or ovalbumin (OVA) adsorbed to alum. Specific antibody isotype responses, T cell proliferation, T cell cytokine production and anaphylaxis were assessed throughout the sensitization phase. RESULTS: Low-dose immunization with PLA2 (0.1 microg) favoured a long-term, specific T helper (Th)2 response with high IgE and IL-4 production in contrast to high-dose PLA2 (10 microg) immunization, which biased the immune response towards a Th1 response with high IgG2a and low IL-4 production. Parallel immunization with an unrelated antigen (ovalbumin) had a significant bystander effect on the immunization with PLA2, which was also dose-dependent. Finally, although anaphylaxis as measured by rectal temperature drop was allergen-specific, it could be induced in the high- and low-dose immunization groups, and was not solely dependent on IgE levels. CONCLUSION: Though low-dose allergen immunization appears to induce an efficient IgE response, the intensity and quality of this response may be modulated by bystander effects of parallel immunization and does not correlate strictly with anaphylaxis. This observation has relevance to the design of clinical immunotherapy protocols using murine model-based data. PMID- 11940073 TI - Cytokine pattern in allergic and non-allergic chronic rhinosinusitis in asthmatic children. AB - BACKGROUND: Rhinosinusitis represents one of the most common chronic diseases. The association of rhinosinusitis with asthma has been frequently reported. Eosinophils and Th2 cells play a pathogenic mechanism in asthma. OBJECTIVE: The aims of the study were to evaluate the cytokine pattern in chronic rhinosinusitis in asthmatic children and to compare the findings in allergic vs. non-allergic asthmatics. METHODS: Thirty-five asthmatic children were evaluated, 19 males and 16 females, with an average age of 8.7 years. All children were asthmatic and suffered from chronic rhinosinusitis. Twenty were allergic and 15 were non allergic. Ten healthy children were studied as normal controls. Evaluated parameters were the levels of the following cytokines: IL-1beta, IL-4, IL-6, IL 8, IL-12, IFN-gamma and TNF-alpha. Cytokines were recovered from rhinosinusal lavage and measured by immunoassays. Nasal cytology was also performed in all subjects and inflammatory cells were counted by conventional staining. RESULTS: Allergic subjects showed a significant increase of IL-4 (P < 0.01) and TNF-alpha (P < 0.05) and a significant decrease of IL-12 (P < 0.05) and of IFN-gamma (P < 0.0001), whereas IL-1beta, IL-6 and IL-8 were not significantly increased. Non allergic children showed a significant increase of IL-4 (P < 0.05) and a significant decrease of IFN-gamma (P < 0.0001), IL-12 was not significantly decreased, and IL-1beta, IL-6 and IL-8 were not significantly increased. A significant inflammatory infiltrate was present in all asthmatic children. Significant correlations were demonstrated between IL-4 and IL-12 (P < 0.001), IL 12 and IFN-gamma (P < 0.001), IL-8 and neutrophils (P < 0.01), and TNF-alpha and monocytes/macrophages (P < 0.05), in allergic asthmatics. IL-4 and IL-12 were significantly correlated (P < 0.05) as well as IL-8 and neutrophils (P < 0.01) in non-allergic asthmatics. CONCLUSION: This study shows that allergic asthmatic children with chronic rhinosinusitis have a typical Th2 cytokine pattern, but also non-allergic asthmatic children share a similar pattern. These findings would suggest the existence of a common pathophysiological mechanism shared by upper and lower airways and are consistent with the concept of united airways disease. PMID- 11940072 TI - Inhalation of a harmless antigen (ovalbumin) elicits immune activation but divergent immunoglobulin and cytokine activities in mice. AB - BACKGROUND: Exposure to aerosolized harmless antigen such as ovalbumin (OVA) has previously been shown to induce inhalation tolerance, a state characterized by inhibition of IgE synthesis and airway inflammation, upon secondary immunogenic antigen encounter. Immune events associated with this phenomenon are still poorly understood. OBJECTIVE: The aim of this study was to investigate cellular and molecular mechanisms underlying this state of 'unresponsiveness'. METHODS: After initial repeated OVA exposure, mice were subjected to a protocol of antigen induced airway inflammation, encompassing two intraperitoneal injections of OVA adsorbed to aluminium hydroxide followed by airway challenge. We assessed immune events in the draining lymph nodes after sensitization, and in the lungs after challenge. RESULTS: In animals initially exposed to OVA, we observed, at the time of sensitization, considerable expansion of T cells, many of which expressed the activation markers CD69 and CD25, as well as increased numbers of antigen presenting cells, particularly B cells. While these animals produced low levels of IgE, the observed elevated levels of IgG1 signified isotype switching. Splenocytes and lymph node cells from OVA-exposed mice produced low levels of IL 4, IL-5, IL-13 and IFN-gamma, indicating aborted effector function of both T helper (Th)2- and Th1-associated cytokines. Real time quantitative polymerase chain reaction (PCR) (TaqMan) analysis of costimulatory molecules in the lungs after in vivo challenge showed that B7.1, B7.2, CD28 and CTLA-4 mRNA expression was low in animals initially exposed to OVA. Ultimately, these events were associated with abrogated airway inflammation and attenuated airway hyper responsiveness. The decreased inflammation was antigen-specific and independent of IL-10 or IFN-gamma. CONCLUSION: Initial exposure to OVA establishes a programme that prevents the generation of intact, fully functional inflammatory responses upon secondary antigen encounter. The absence of inflammation, however, is not associated with categorical immune unresponsiveness. PMID- 11940074 TI - Peripheral blood CD4+ and CD8+ T cell type 1 and type 2 cytokine production in atopic asthmatic and normal subjects. AB - BACKGROUND: Increased production of IL-4 and IL-5 and decreased production of IFN gamma by CD4+ T cells has been implicated in asthma pathogenesis. However, CD8+ T cells also produce type 1 and type 2 cytokines and the relative roles of CD4+ and CD8+ T cell cytokine production in asthma have not been previously studied. OBJECTIVE: To determine the production of the type 1 and type 2 cytokines by CD4+ and CD8+ T cell subsets in asthmatic and normal subjects. METHODS: Intracellular cytokine staining for IL-4, -5, -10, -13 and IFN-gamma was analysed in peripheral blood CD4+ and CD8+ T cells from 24 atopic asthmatic and 20 normal subjects. RESULTS: Both subsets of T cells produced all cytokines studied and there were no significant differences between CD4+ and CD8+ T cells in their capacity to produce either type 1 or type 2 cytokines. There were significantly increased frequencies of IFN-gamma-positive CD4+ (13.1 +/- 2.4%, vs. 7.3 +/- 1.4%) and CD8+ (20.0 +/- 2.9%, vs. 9.6 +/- 2.1%) T cells in asthmatic subjects compared with normal subjects (P < 0.05), but not in frequencies of CD4+ or CD8+ T cells staining positively for IL-4, -5, -10 or -13. CONCLUSION: The frequencies of peripheral blood CD8+ T cells producing type 1 and type 2 cytokines are comparable with the frequencies of CD4+ T cells. There was an increased frequency of IFN-gamma producing CD4+ and CD8+ T cells in asthmatic compared with normal subjects. Further studies investigating T cells derived from the airways and investigating various stages within the disease process are required to further elucidate the importance of type 2 and type 1 T cell cytokine production in the pathogenesis of human allergic disease. PMID- 11940075 TI - Prevalence of occupational allergy to bell pepper pollen in greenhouses in the Netherlands. AB - BACKGROUND: An increasing number of allergic complaints appear to have occurred among bell pepper greenhouse employees. OBJECTIVE: The aim of this study was to estimate the prevalence of work-related allergic symptoms and the prevalence of sensitization to specific occupational allergens and its determinants. METHODS: We studied 472 employees who were invited to answer an extensive questionnaire and to be tested on location with inhalant allergens and home-made extracts of the bell pepper plant. In addition, peak expiratory flow monitoring and RASTs were performed. RESULTS: Work-related symptoms were reported in 53.8% of all cases. Sensitization to the bell pepper plant was found in 35.4%. Positive reactions to leaf, stem and/or juice, however, were associated in nearly 90% with sensitization to pollen, which appeared to be most important allergen of the plant. Sensitization to the bell pepper plant and inhalant atopy were considered the most important risk factors for the occurrence of work-related symptoms of the upper airways (PRR 2.63, CI 2.11-3.25 and PRR 2.25, CI 1.82-2.79) as well as of the lower airways (PRR 4.08, CI 2.38-7.00 and PRR 3.16, CI 1.87-5.33). CONCLUSION: There is a surprisingly high prevalence of work-related respiratory symptoms (53.8%) in bell pepper horticulture. In two-thirds of the employees, symptoms at work were associated with an IgE-mediated allergy due to the high and chronic exposure to bell pepper pollen. Complaints at work without specific sensitization to bell pepper pollen can be caused by non-specific hyper reactivity or atopy to other occupational allergens. The extent of this occupational allergy has important consequences for the health care of this large, still growing occupational group. PMID- 11940076 TI - Type I allergy to natural rubber latex and type IV allergy to rubber chemicals in health care workers with glove-related skin symptoms. AB - BACKGROUND: It has been established that there are type I and type IV allergens in latex gloves. OBJECTIVE: The purpose of the study was to establish the prevalence of rubber glove-induced skin symptoms among health care workers in one Italian hospital. METHODS: Health care workers (n = 1584) were evaluated using a written questionnaire and 295 respondents with glove-induced skin symptoms were tested. We performed: skin prick test with latex glove extract and commercial latex, and environmental and food allergens; glove use test; patch tests with a rubber additive series; and RASTs. RESULTS: Hospital employees who used or had used latex gloves at work were 1294. Three hundred and sixteen (24.4%) reported glove-induced symptoms, namely, cutaneous symptoms in all the cases and non cutaneous symptoms in 105 subjects (8.1%). Twenty-seven of the 295 symptomatic employees tested (9.1%) were latex sensitive. Thirty-one patients (10.5%) exhibited positive patch test to rubber-related allergens. The most positive readings were obtained from the Thiuram mix and the Carba mix, with 12 and 9 positivities, respectively. The risk factors for latex skin sensitization were: a previous history of atopy and asthma; history of surgery; pre-existing hand dermatitis; work-related symptoms; and positive skin tests to common inhalant and certain foods (P < 0.05). Subjects who exhibited positive patch test were significantly more likely to have a prior hand dermatitis (P < 0.001). Of the 295 cases, 275 (93.2%) were contact dermatitis (CD), 13 (4.4%) contact urticaria (CU) (including protein CD) and 7 (2.4%) CD associated with CU. CONCLUSIONS: Our results show a high prevalence of rubber glove-induced dermatoses among the employees in one Italian hospital. The majority of skin complaints of latex gloves are related to skin irritation rather than to allergy. The immediate allergy to latex and the delayed allergy to rubber chemicals suggest that all the health care workers with glove-related dermatitis should undergo both skin prick test and glove use test to detect type I hypersensitivity to latex, and patch test to detect type IV hypersensitivity to rubber chemicals. PMID- 11940077 TI - What is the role of the hevein-like domain of fruit class I chitinases in their allergenic capacity? AB - BACKGROUND: Class I chitinases are the major panallergens in fruits associated with the latex-fruit syndrome. These enzymes contain an N-terminal hevein-like domain homologous to latex hevein, and a larger catalytic domain. The role of these domains in their allergenic capacity is still controversial. OBJECTIVE: We sought to evaluate the role of both domains of class I chitinases in their IgE binding properties, using Cas s 5, the major allergen from chestnut, as a model. METHODS: Recombinant Cas s 5 and its deleted form, lacking the hevein-like domain, designated rCat, were expressed in Pichia pastoris using the pPIC 9 vector. Both recombinant products were purified from the supernatants of transformed yeast cultures by gel-filtration and cation-exchange chromatography. The isolated proteins were characterized by N-terminal sequencing, enzymatic activity and N-glycosylation tests, anti-chitinase and specific IgE immunodetection. Immunoblot, RAST and CAP inhibition assays were also performed. RESULTS: Both purified rCas s 5 and rCat showed the expected N-terminal amino acid sequences and an enzymatic activity similar to that of their natural counterparts isolated from chestnut seeds, and were strongly recognized by anti chitinase antibodies. In contrast, only rCas s 5, but not rCat, bound specific IgE from sera of patients suffering from the latex-fruit syndrome, and fully inhibited IgE-binding to natural Cas s 5 in immunoblot inhibition assays. Latex hevein also exerted a strong immunoblot inhibition of IgE-binding to chestnut Cas s 5. RAST and CAP inhibition using whole chestnut extract on the solid phase, rendered inhibition levels around 70-90% for rCas s 5 and 60% for rCat, in contrast to the immunoblotting results. CONCLUSIONS: Recombinant Cas s 5 behaves like natural Cas s 5 in IgE-binding assays in vitro. The hevein-like domain of allergenic class I chitinases seems to include all their main IgE-binding epitopes when tested by immunodetection and immunoblot inhibition experiments. RAST and CAP inhibition assays, on the contrary, suggest that relevant epitopes are also harboured in the catalytic domain of these allergens. PMID- 11940078 TI - Cloning and molecular characterization of the Hevea brasiliensis allergen Hev b 11, a class I chitinase. AB - BACKGROUND: In the last 10 years type-I allergy against proteins from Hevea brasiliensis latex has become an acknowledged medical issue. Fruit-allergic patients represent one risk group for developing latex allergy. Class I chitinases have been identified from chestnut, avocado and banana as relevant allergens. The chitin binding (hevein) domain from these class I chitinases has been postulated to bear the important IgE binding epitopes. OBJECTIVE: To clone the cDNA of an allergenic latex class I chitinase, to express the recombinant protein and to determine its IgE cross-reactivity with hevein (Hev b 6.02). METHODS: A full-length cDNA coding for a class I chitinase has been isolated from Hevea latex RNA by reverse transcription followed by PCR. The chitinase encoding sequence has been subcloned into the pMAL expression vector and expressed in E. coli as a fusion protein to maltose binding protein. The highly enriched recombinant protein fraction has been tested for its IgE binding capacity in immunoblots and ELISA. Furthermore, the pathogenesis-related function of the recombinant protein was tested in a fungal growth inhibition assay. RESULTS: The Hevea brasiliensis latex chitinase, designated Hev b 11, displays 70% identity to the endochitinase from avocado and its hevein-domain 58% to hevein (Hev b 6.02). The recombinant Hev b 11-maltose binding protein is recognized by latex- and fruit-allergic patients with IgE binding in both, ELISA and immunoblots. Pre incubation of sera with rHev b 11-maltose binding protein showed an overall 16% inhibition of subsequent binding to rHev b 6.02-maltose binding protein on solid phase. The growth of F. oxysporum was inhibited in a dose dependent manner by addition of rHev b 11-maltose binding protein to the culture. CONCLUSIONS: Hev b 11, a class I chitinase, is another allergen from Hevea latex with a chitin binding domain and displays a different IgE binding capacity compared with hevein. PMID- 11940079 TI - Influence of respiratory syncytial virus infection on cytokine and inflammatory responses in allergic mice. AB - BACKGROUND: Th2 lymphocyte responses are associated with inflammation and disease during allergic responses. Exposure to particular environmental factors during the expression of allergy could result in more pronounced Th2-like immune responses and more severe disease. One factor might be a respiratory virus infection. OBJECTIVE: The aim of our study was to investigate the influence of respiratory syncytial virus (RSV) infection on the expression of ovalbumin (OVA) induced allergy in BALB/c mice. METHODS: We determined OVA-specific IgE in serum, cytokine profiles and histopathological lesions in lungs of OVA-allergic mice after RSV infection. RESULTS: OVA sensitization and challenge induced OVA specific IgE in serum, Th2 cytokine mRNA expression, and mononuclear and eosinophilic inflammation in the lungs. RSV inoculation during the challenge period enhanced OVA-induced IL-4 and IL-5 mRNA expression in lung tissue. RSV further enhanced the OVA-induced hypertrophy of mucous cells and eosinophilic infiltration in lung tissue. Surprisingly, RSV infection decreased Th2 cytokine secretion and eosinophilic influx in bronchoalveolar lavage of OVA-allergic mice. Because inactivated RSV did not influence these responses, replication of RSV appeared essential for the modification of OVA-induced Th2 cytokine expression. RSV did not change OVA-specific IgE levels in serum. Furthermore, the RSV-induced IL-12 mRNA expression in lung tissue of OVA-allergic mice was diminished, but IFN gamma mRNA expression was not affected. CONCLUSION: RSV infection enhanced particular OVA-induced Th2 cytokine mRNA responses and pulmonary lesions in allergic mice and thus aggravated allergic respiratory disease. PMID- 11940080 TI - The involvement of sensory neuropeptides in airway hyper-responsiveness in rabbits sensitized and challenged to Parietaria judaica. AB - BACKGROUND: C-fibres have received considerable attention in the context of airway hyper-responsiveness (AHR), in fact several lines of evidence suggest that tachykinins might be involved in the pathogenesis of AHR. OBJECTIVE: The aim of this study was to investigate the role of capsaicin-sensitive sensory C-fibres and tachykinins in rabbits sensitized to the major allergen of Parietaria judaica pollen (Par j1). METHODS: Airway responsiveness was determined by exposing sensitized rabbits to cumulative concentrations of aerosolized histamine before and after an allergic challenge and after a pre-treatment with either vehicle or capsaicin or tachykinin receptor antagonists. Bronchoalveolar lavage was performed following histamine challenge and total and differential cell counts were performed. RESULTS: In sensitized rabbits, an AHR to inhaled histamine was observed 24 h after a Par j1 challenge. Capsaicin pre-treatment inhibited the AHR achieved 24 h following antigen exposure (P < 0.01). Pre-treatment with the tachykinin NK2 receptor antagonist, SR 48968, significantly reduced the antigen induced AHR (P < 0.05), while pre-treatment with tachykinin NK1 (SR 140333) and NK3 (SR 142801) receptor antagonists did not significantly modify it. Bronchoalveolar lavage fluid obtained from vehicle and capsaicin-treated rabbits challenged with Par j1 exhibited no significant differences in total and differential cell counts. CONCLUSIONS: Parietaria judaica-induced AHR in immunized rabbits was shown to be inhibited by pre-treatment with capsaicin, an effect that is not related to an action on the associated pulmonary infiltration of eosinophils. The involvement of NK2 receptor stimulation in this phenomenon also suggests that NK2 receptor antagonists may be useful for investigating mechanisms of bronchopulmonary alterations in asthmatic patients. PMID- 11940081 TI - The complexities of ocular genetics. AB - The cliche 'a picture is worth a thousand words' is a testament to the power of the visual system in helping us deal with our physical environment. Rarely do perturbations to the visual system, even minor ones, go unnoticed. Major defects in eye development may occur in the absence of systemic problems which threaten health. Ocular anomalies offer a window into many developmental events which would otherwise be difficult to study. PMID- 11940082 TI - Genetic control of caudal development. AB - Several lines of evidence suggest that caudal development involves a distinct programme. This is illustrated by the fact that a specific pattern of malformations affects the caudal end of the human embryo. In addition, neurulation, the process leading to the formation of the neural tube, proceeds through different morphogenetic movements caudally. In mammals, as in birds, the caudal neural tube arises from cavitation and not from folding of the neural plate as in more rostral levels. However, recent fate mapping studies have suggested that the two modes of neurulation represent a continuous programme, possibly involving similar cellular or molecular mechanisms. Finally, analyses of mutant mice have shown that T-box transcription factors and components of the Wnt signalling pathway control cellular migration and the promotion of mesoderm formation in the caudal embryo. In humans, mutation in the HLXB9 transcription factor causes an autosomal dominant form of sacral agenesis. Thus, the combination of classical embryological and molecular genetics approaches has provided critical reference points for the delineation of the developmental programme of the caudal embryo. PMID- 11940084 TI - Acanthocytosis in a patient with homozygous familial hypobetalipoproteinemia due to a novel APOB splice site mutation. AB - We report on acanthocytosis in a 31-year-old woman with homozygous familial hypobetalipoproteinemia due to a mutation affecting the splicing of the APOB gene encoding apolipoprotein B. Treatment with fat-soluble vitamins was associated with arrest of the usually progressive neurological complications of this condition. However, the acanthocytosis - literally 'thorny' erythrocytes that result from abnormal membrane fluidity - persists despite treatment. PMID- 11940085 TI - Participation rates of Ashkenazi Jews in a colon cancer community-based screening/prevention study. AB - In a recent colon cancer risk study, genetic assessment and colonoscopy were offered to virtually all of the adult Ashkenazi Jews in an urban community. The present study was designed to examine factors influencing participation and response in the initial study and to suggest strategies for improving participation in future health promotion programs. The study comprised a random sample of three groups of individuals who had been targeted for participation in the previous study: those who had (a) agreed to participate (n = 234); (b) declined participation (n = 179); and (c) failed to respond to a mailed recruitment package (n = 128). All participants completed a brief telephone survey. Key multivariate predictors of both response and participation were individuals' perceptions of the drawbacks of participating in colon cancer screening research and the degree of decisional conflict they experienced. Response was further predicted by the influence of spouses, family history of colon cancer, past knowledge of genetic testing for colon cancer, and education level. Participation was predicted by awareness that the study was supported by the Ashkenazi Jewish community, past experience with genetic testing, individuals' perceptions of the benefits of participating, and whether or not they had children. The degree to which individuals understand the purpose and nature of genetic screening research, along with their levels of decisional conflict and other psychosocial factors, may influence the likelihood of their participation in such research. Results of this study suggest a number of possible strategies for improving participation and response rates in disease prevention and detection studies. PMID- 11940087 TI - The effect of six polymorphisms in the Apolipoprotein B gene on parameters of lipid metabolism in a Danish population. AB - Lipoproteins are vehicles for the distribution of plasma lipids and polymorphisms in the genes for apolipoproteins could influence the amount of lipid in plasma. We examined the effect of six single nucleotide polymorphisms in codons 71, 591, 2488, 2712, 3611, and 4154 of the apolipoprotein B gene on fasting levels of triglyceride, VLDL-, LDL-, HDL- and total cholesterol and on body mass index (BMI) in a cohort of 2656 Danes aged 40-70 years using a linear model correcting for the effects of gender, age, BMI, smoking, alcohol consumption and physical activity. The codon 2488 polymorphism was the most influential of the tested polymorphisms, significantly influencing triglyceride (P = 0.002), LDL cholesterol (P < or = 0.0004), VLDL-cholesterol (P = 0.006) and total cholesterol (P = 0.0001). The codon 2712 polymorphism had an impact on triglyceride (P = 0.007) and VLDL-cholesterol (P = 0.001), while the codon 71 polymorphism influenced LDL- and total cholesterol (P = 0.04 and P = 0.02, respectively). An interaction between smoking and codon 591 (P = 0.03) and smoking and codon 3611 (P = 0.02) on BMI was observed, as well as modest interactions between codon 3611 and codons 2488 and 2712 on lipid parameters. All polymorphisms were in close linkage disequilibrium. The population was not in Hardy-Weinberg equilibrium in four of the six polymorphisms but the lack of equilibrium was restricted mainly to the 60-year olds. PMID- 11940088 TI - A CGH study of 27 patients with CHARGE association. AB - CHARGE association is a non-random occurrence of congenital malformations including coloboma, heart disease, choanal atresia, retarded growth and/or retarded development, genital hypoplasia, ear anomalies and/or deafness. The cause of this association remains unknown. Various genetic mechanisms have been proposed, including a contiguous gene syndrome but, so far, no recurrent locus has been identified. To address this question, we decided to perform a comparative genomic hybridization (CGH) study on a cohort of 27 patients with CHARGE association and a normal standard karyotype. We found two chromosomal anomalies: a der(9)t(9;13) derived from a paternal translocation and a der(6)t(4;6) of unknown origin. This suggests that chromosome imbalances may well mimic CHARGE association. Therefore patients with CHARGE association must be carefully tested with classical and molecular cytogenetic techniques to detect a potential chromosome imbalance. It is expected that more stringent diagnostic criteria of CHARGE association could define a more homogeneous group of patients where a single genetic cause might be identified. PMID- 11940086 TI - ABCA1 regulatory variants influence coronary artery disease independent of effects on plasma lipid levels. AB - The authors have previously shown that individuals heterozygous for ABCA1 mutations have decreased high density lipoprotein cholesterol, increased triglycerides and an increased frequency of coronary artery disease (CAD), and that single nucleotide polymorphisms (SNPs) in the coding region of the ABCA1 gene significantly impact plasma lipid levels and the severity of CAD in the general population. They have now identified several SNPs in non-coding regions of ABCA1 which may be important for the appropriate regulation of ABCA1 expression (i.e. in the promoter, intron 1 and the 5' untranslated region), and have examined the phenotypic effects of these SNPs in the REGRESS population. Out of 12 SNPs, four were associated with a clinical outcome. A threefold increase in coronary events with an increased family history of CAD was evident for the G 191C variant. Similarly, the C69T SNP was associated with a twofold increase in events. In contrast, the C-17G was associated with a decrease in coronary events and the InsG319 was associated with less atherosclerosis. For all these SNPs, the changes in atherosclerosis and CAD occurred without detectable changes in plasma lipid levels. These data suggest that common variation in non-coding regions of ABCA1 may significantly alter the severity of atherosclerosis, without necessarily influencing plasma lipid levels. PMID- 11940089 TI - Non-syndromic X-linked mental retardation associated with a missense mutation (P312L) in the FGD1 gene. AB - Three brothers with non-syndromal X-linked mental retardation were found to have a novel missense mutation in FGD1, the gene associated with the Aarskog syndrome. Although the brothers have short stature and small feet, they lack distinct craniofacial, skeletal or genital findings suggestive of Aarskog syndrome. Their mother, the only obligate carrier available for testing, has the FGD1 mutation. The mutation, a C934T base change in exon 4, results in the proline at position 312 to be substituted with a leucine. This missense mutation is predicted to eliminate a beta-turn, creating an extra-long stretch of coiled sequence which may affect the orientations of an SH3 (Src homology 3) binding domain and the first structural conserved region. A new molecular defect associated with non syndromal X-linked mental retardation affords an opportunity to seek specific diagnosis in males with previously unexplained developmental delays and this opens further predictive tests in families at risk. PMID- 11940090 TI - RMRP gene sequence analysis confirms a cartilage-hair hypoplasia variant with only skeletal manifestations and reveals a high density of single-nucleotide polymorphisms. AB - Mutations in the RMRP gene that codes for an RNA subunit of the MRP RNAse complex are the cause of cartilage-hair hypoplasia (CHH; MIM 250250). We tested the hypothesis that recessive metaphyseal dysplasia without hypotrichosis (M1M 250460), a disorder presenting with short stature and metaphyseal dysplasia similar to CHH, but lacking hair anomalies, immunodeficiency and other extra skeletal features, might be allelic to CHH. We identified four mutation-carrying alleles segregating with the skeletal phenotype in two unrelated boys and their parents. One allele carried the common Finnish mutation +70A--> G; the remaining three carried +195C--> T, +238C--> T, and dupAAGCTGAGGACG at -2. Sequencing 120 alleles from a control group revealed an unusually high density of single nucleotide polymorphisms in and around the RMRP gene: the biological significance of this finding is unclear. We conclude that recessive metaphyseal dysplasia without hypotrichosis is a variant of CHH, manifesting only as short stature and metaphyseal dysplasia. Precise diagnosis of this form of metaphyseal dysplasia is not without importance because of recessive inheritance with corresponding recurrence risk, as well as because of potential complications such as anaemia, susceptibility to infections and the increased likelihood of developing cancer. The short stature and metaphyseal changes associated with cone-shaped epiphyses of the hands should raise the diagnostic possibility of a CHH-related disorder that can then be confirmed by mutation analysis. PMID- 11940091 TI - A patient with treatment-resistant schizophrenia and cytochrome P4502D6 gene duplication. AB - We describe a patient with treatment-resistant schizophrenia who had a duplication in the cytochrome P450IID6 (CYP2D6) gene. This severely ill 71-year old-woman had responded poorly to several neuroleptics. Molecular genetic study revealed CYP2D6 gene duplication, which results in excessive activity of CYP2D6 that metabolizes various commonly used neuroleptics. The mutation may have contributed to treatment resistance in this case. PMID- 11940092 TI - A low prevalence of the C677T mutation in the methylenetetrahydrofolate reductase gene in Asian Indians. AB - The prevalence of the C677T mutation in the methylenetetrahydrofolate reductase (MTHFR) gene in Asian Indians from India was determined and the association of the mutant allele with coronary artery disease (CAD) was evaluated in a case control study. The case group consisted of 251 patients with CAD; 195 male and 56 female aged from 29 to 82 years (mean age +/- SD, 57.5 +/- 10.6 years). The control group consisted of 216 apparently healthy individuals without evidence of CAD; 161 male and 55 female aged from 30 to 83 years (mean age +/- SD, 54.9 +/- 10.4 years). All the patients were assessed by coronary angiography. While 33 patients had normal coronaries, 23, 25 and 39 patients had single-vessel, two vessel and triple-vessel disease, respectively. Eighty-three patients (33%) had suffered myocardial infarction less than a year to five years earlier. The C677T polymorphism in the MTHFR gene was assessed. While 31% of the controls and 38% of the patients had the heterozygous genotype, 2% of the control group and none of the patients had the mutant homozygous genotype. The overall 'T' allelic frequencies were comparable in control and patient groups (0.18 and 0.19, respectively), but the association of the sum of heterozygous and homozygous genotypes with CAD (1, 2 or 3-vessel disease) was statistically significant for females only [Odds ratio (95% confidence intervals), 2.8 (1.1-6.9), p = 0.023]. No association was found between genotype distribution and previous myocardial infarction or severity of atherosclerosis. PMID- 11940093 TI - Exclusion of candidate gene loci for adult onset primary open-angle glaucoma in a genetically isolated population. PMID- 11940094 TI - Molecular cytogenetic analysis of a Xp21.3-pter deletion in a family with normal and short stature. PMID- 11940096 TI - Mutations of stonewall disrupt the maintenance of female germline stem cells in Drosophila melanogaster. AB - Germline stem cells located at the anterior tip of the adult Drosophila melanogaster ovary are critical to the continuous production of mature eggs. Following germline stem cell division, one daughter cell remains a stem cell, while the other becomes a cystoblast committed to differentiation. In this study it was shown that mutations in the putative transcription factor stonewall (stwl) disrupted the maintenance of female germline stem cells. The stwl mutations resulted in a loss of germline stem cells, causing a rapid decrease in egg chamber production. The egg chambers developed only to a limited extent before degenerating. The four mitotic cystocyte divisions were frequently inhibited by stwl mutations. Furthermore, some stwl germaria from newly emerged females completely lacked both stem cells and developing cysts and had a strong reduction in size. The argument is presented here that stwl is involved in the continuation of cell division during female germline development. PMID- 11940097 TI - Synthesis and release of activin and noggin by cultured human amniotic epithelial cells. AB - Recent studies suggest that extra-embryonic tissues may be essential sources of early organizing signals for the mouse embryo. In vitro studies of human amniotic epithelial cells (HAEC) have shown that the amnion can produce various biologically active substances. In this study, the synthesis and release of activin A and noggin, and the activin signaling pathway, was investigated in HAEC. Conditioned medium from cultured HAEC contained activin A which was functionally active in Xenopus laevis animal cap assays. Immunohistochemistry, western blotting and reverse transcription-polymerase chain reaction confirmed that HAEC also synthesize and release noggin. Noggin transcripts were induced by the addition of recombinant activin A, and activin A was inhibited by activin antibody except in the presence of cycloheximide (CHX). These data demonstrate that noggin mRNA expression is induced directly by activin A without new protein synthesis, indicating that noggin is a primary response gene. The results suggest that there is an activin signaling pathway in HAEC, and that the human amnion might therefore be involved in neural formation during early development. PMID- 11940098 TI - Process of pigment cell specification in the sand dollar, Scaphechinus mirabilis. AB - The process of pigment cell specification in the sand dollar Scaphechinus mirabilis was examined by manipulative methods. In half embryos, which were formed by dissociating embryos at the 2-cell stage, the number of pigment cells was significantly greater than half the number of pigment cells observed in control embryos. This relative increase might have been brought about by the change in the arrangement of blastomeres surrounding the micromere progeny. To examine whether such an increase could be induced at a later stage, embryos were bisected with a glass needle. When embryos were bisected before 7 h postfertilization, the sum of pigment cells observed in a pair of embryo fragments was greater than that in control embryos. This relative increase was not seen when embryos were bisected after 7 h postfertilization. From the size of blastomeres, it became clear that the 9th cleavage was completed by 7 h postfertilization. Aphidicolin treatment revealed that 10-15 pigment founder cells were formed. The results obtained suggest that the pigment founder cells were specified through direct cell contact with micromere progeny after the 9th cleavage, and that most of the founder cells had divided three times before they differentiated into pigment cells. PMID- 11940099 TI - Study of Cynops pyrrhogaster notochord differentiation using a novel monoclonal antibody. AB - Two monoclonal antibodies which reacted specifically with the notochord of the early Cynops pyrrhogaster embryo were screened. The antigen molecules were detected within and around the notochord. They were first found mostly between the neural plate and the dorsal part of the notochord in the early neurula (stage 15). They were subsequently detected between the notochord and the somite in the advanced embryo, and they were last detected between the notochord and the underlying endoderm. Whole-mount labeling indicated that the antigen molecules were first detected in the anterior half of the notochord in the early neurula (stage 15). The signals gradually spread along the anterior-posterior axis, especially towards the posterior region. This fact suggests that notochord differentiation progresses from the anterior region which first receives the dorsal mesoderm-inducing signals released horizontally from the lower dorsal marginal zone during early gastrulation. The present study suggested that: (i) notochord differentiation proceeds from the anterior region; and (ii) secretion of the antigen molecules results in the drawing of a boundary between the adjacent tissues. PMID- 11940100 TI - Dissecting planarian central nervous system regeneration by the expression of neural-specific genes. AB - The planarian central nervous system (CNS) can be used as a model for studying neural regeneration in higher organisms. Despite its simple structure, recent studies have shown that the planarian CNS can be divided into several molecular and functional domains defined by the expression of different neural genes. Remarkably, a whole animal, including the molecularly complex CNS, can regenerate from a small piece of the planarian body. In this study, a collection of neural markers has been used to characterize at the molecular level how the planarian CNS is rebuilt. Planarian CNS is composed of an anterior brain and a pair of ventral nerve cords that are distinct and overlapping structures in the head region. During regeneration, 12 neural markers have been classified as early, mid regeneration and late expression genes depending on when they are upregulated in the regenerative blastema. Interestingly, the results from this study show that the comparison of the expression patterns of different neural genes supports the view that at day one of regeneration, the new brain appears within the blastema, whereas the pre-existing ventral nerve cords remain in the old tissues. Three stages in planarian CNS regeneration are suggested. PMID- 11940101 TI - Antisense inhibition of Xbrachyury impairs mesoderm formation in Xenopus embryos. AB - Expression of the Xbrachyury (Xbra) gene was inhibited by antisense RNA synthesized in situ from an expression vector read by RNA polymerase III, injected into the fertilized egg or the 2-cell stage embryo of Xenopus laevis. Antisense-treated embryos had markedly reduced levels of Xbra mRNA and protein, and showed deficiencies in mesodermal derivatives and axis formation. In particular, organization of the posterior axis was affected, but often the anterior axis was also reduced. Some embryos failed to form mesoderm altogether and remained amorphous. The antisense effect is dose-dependent and may be "rescued" by overexpression of Xbra. In Xbra-deficient embryos, expression of several mesodermal genes (Xvent, pintallavis, Xlim, Xwnt-8 and noggin) was reduced to varying degrees, whereas goosecoid levels remained normal. The modified expression levels were partly normalized when Xbra deficiency was rescued. The observation that antisense inhibition yields slightly different phenotypes from dominant-negative inhibition suggests the recommendation of using several surrogate genetic approaches to determine the functional role of a gene in Xenopus development. PMID- 11940102 TI - In vitro induction of the pronephric duct in Xenopus explants. AB - The earliest form of embryonic kidney, the pronephros, consists of three components: glomus, tubule and duct. Treatment of the undifferentiated animal pole ectoderm of Xenopus laevis with activin A and retinoic acid (RA) induces formation of the pronephric tubule and glomus. In this study, the rate of induction of the pronephric duct, the third component of the pronephros, was investigated in animal caps treated with activin A and RA. Immunohistochemistry using pronephric duct-specific antibody 4A6 revealed that a high proportion of the treated explants contained 4A6-positive tubular structures. Electron microscopy showed that the tubules in the explants were similar to the pronephric ducts of normal larvae, and they also expressed Gremlin and c-ret, molecular markers for pronephric ducts. These results suggest that the treatment of Xenopus ectoderm with activin A and RA induces a high rate of differentiation of pronephric ducts, in addition to the differentiation of the pronephric tubule and glomus, and that this in vitro system can serve as a simple and effective model for analysis of the mechanism of pronephros differentiation. PMID- 11940103 TI - Post-nodal mesoblast caudalizes the host axis and inhibits cell population growth, and induces new primitive streaks in chick embryos. AB - One to eight post-nodal fragments (PN) or Hensen's nodes (HN) from full primitive streak stage chick embryos were transplanted onto the area pellucida or area opaca of stage 4 embryos and cultured for 20 h. Thirteen morphological and numerical parameters were affected in the host embryos and analyzed by multiple logistic regression for parametric hierarchy. In the area pellucida, both PN and HN transplants inhibited cell population growth while only PN caudalized the host axis and induced supernumerary primitive streaks expressing the mesoderm-specific gene Brachyury. In the area opaca, neither grafts influenced host axis morphogenesis, but PN inhibited the cell population growth significantly. Tracking [(3)H]TdR labeled grafts showed that PN cells migrated towards the host axis and participated in the formation of supernumerary somites and hearts. When placed near the host axis, PN caudalized it and inhibited cell population growth. PMID- 11940104 TI - Blood pressure control in patients with type 2 diabetes and hypertension. PMID- 11940105 TI - Vasopeptidase inhibitors and their potential role in diabetes. PMID- 11940106 TI - Identification of two novel and potent competitive inhibitors of the glucose-6 phosphatase catalytic protein. AB - AIM: In this study, we show that inhibitors of the glucose-6-phosphatase (G-6 Pase) catalytic protein could be an alternative approach to the recent G-6-Pase T1-translocase inhibitors to target this enzyme for the treatment of type 2 diabetes. METHOD: The active enantiomers of 4-methoxyphenyl-[4-(4-methoxyphenyl) 4,5,6,7-tetrahydrothieno[3,2-c]pyridin-5-yl]methanone (Compound A-1) and 4 methoxyphenyl-[4-(4-trifluoromethoxyphenyl)-4,5,6,7-tetrahydrothieno[3,2 c]pyridin-5-yl]methanone (Compound B-1) were characterized as inhibitors of the G 6-Pase catalytic protein using pig and rat liver microsomes and cultured rat hepatocytes. RESULTS: Both compounds were found to be potent competitive inhibitors of the G-6-Pase catalytic protein obtained from pig and rat liver microsomes. The K(i) values (microM) were calculated to be 0.61 +/- 0.02 and 0.63 +/- 0.08 for compound A-1 and B-1 on intact pig microsomes, and 0.27 +/- 0.02 and 0.29 +/- 0.06 on disrupted pig microsomes. The corresponding values for rat liver microsomes were found to be 3.3 +/- 0.6 and 4.0 +/- 1.2 for compound A-1 and B-1 on intact microsomes, and 1.54 +/- 0.1 and 1.21 +/- 0.1 on disrupted microsomes. Compound A-1 was also able to inhibit pyrophosphatase activities from both intact and disrupted microsomes with equal potency (IC50; 0.43-0.55 microm). Using cultured rat hepatocytes and glycerol as the substrate, these compounds were able to prevent glucose production up to 60% with a concomitant increase in the G-6-P content (2.3-fold) using compound A-1. No increase in glycogen levels was seen. CONCLUSION: These data demonstrated that these compounds were more potent inhibitors on G-6-Pase obtained from pig microsomes and were able to penetrate the microsomal membrane. The hepatocyte data further support the kinetic data, and are also consistent with the evoked mechanism of action. PMID- 11940107 TI - Does insulin lispro preserve the physiological defences to hypoglycaemia during intensive insulin therapy with a conventional basal bolus regimen? AB - AIM: Insulin lispro used in an intensive basal/bolus regimen produces equivalent glycaemic control to human-soluble insulin but reduces rates of hypoglycaemia. We tested the hypothesis that the use of rapid-acting analogues might prevent the development of defective hypoglycaemic counterregulation during intensive insulin therapy. METHODS: Ten patients with type 1 diabetes (four female, mean age 33 +/- 3 years, diabetes duration 12 +/- 2 years) participated in an open, randomized cross-over study, with 2 months run-in and 4-month treatment periods using either lispro or human-soluble insulin before meals and human NPH insulin (NPH) at night. The total of reported hypoglycaemic episodes (lispro vs. soluble, 123 vs. 128) and HbA(1c) (6.1 +/- 0.2 vs. 6.6 +/- 0.2%) were similar during both treatments. At the end of each period, we measured symptomatic, counterregulatory and cognitive responses, and glycaemic thresholds during hypoglycaemia, induced with a hyperinsulinaemic clamp (blood glucose of 5, 4.5, 3.5 and 2.5 mmol/l). RESULTS: We found similar overall responses of adrenaline, cortisol, growth hormone and total symptom score. Glycaemic thresholds for rises in adrenaline (3.1 +/- 0.2 vs. 3.1 +/- 0.2 mmol/l, p = 0.76), cortisol (2.2 +/- 0.1 vs. 2.2 +/- 0.1 mmol/l, p = 0.16), growth hormone (3.3 +/- 0.15 vs. 2.9 +/- 0.2 mmol/l, p = 0.13), symptoms (3.2 +/- 0.2 vs. 3.3 +/- 0.1 mmol/l, p = 0.051) and impaired cognitive function (3.0 +/- 0.2 vs. 3.0 +/-0.2 mmol/l, p = 0.20) were also similar. CONCLUSION: Four months of intensive treatment, with insulin lispro used pre-prandially and isophane at night, produced relatively preserved but equivalent physiological responses to hypoglycaemia as those on soluble insulin. Longer periods of treatment or alternative regimens may be necessary to demonstrate beneficial effects on hypoglycaemic physiological responses. PMID- 11940108 TI - Obesity is not a confounding factor for performing autonomic function tests in individuals with diabetes mellitus. AB - AIM: Cardiovascular autonomic neuropathy is a serious complication of diabetes mellitus. Previous studies have revealed conflicting results with regard to the role of obesity and its effect on the performance of tests (e.g. RR-variation during deep breathing) for the determination of the presence of cardiovascular autonomic dysfunction. The objective of this study was to determine if obesity affects the performance and the reproducibility of autonomic function tests. METHODS: This cross-sectional study included 159 diabetic individuals. Autonomic function tests included: RR-variation during deep breathing and the Valsalva ratio. These tests were assessed using the ANS2000 ECG Monitor and Respiration Pacer. RR-variation was measured by vector analysis (i.e. mean circular resultant, MCR). Reproducibility of the autonomic function tests was assessed by determining the coefficient of variation (CV) on repeat testing. RESULTS: Using cut-off points to describe normal weight (body mass index (b.m.i.) < or = 25 kg/m(2)), overweight (b.m.i. 25.01-30 kg/m(2)), obese (b.m.i. 30.01-40 kg/m(2)), and morbidly obese (b.m.i. > or = 40.1 kg/m(2)), no difference was found for the MCR, Valsalva ratio, CV of the MCR, or CV of the Valsalva ratio among the various weight levels for individuals with type 1 or type 2 diabetes. CONCLUSIONS: The results of this study indicate that obesity is not a confounding factor in the performance of autonomic function tests. Likewise, the reproducibility of autonomic function testing is not affected by obesity. Assessment of autonomic function is important for obese and non-obese individuals given that reduced RR variation is associated with exercise intolerance, intraoperative cardiovascular lability and increased risk of mortality. PMID- 11940109 TI - Serum homocysteine concentration is related to diabetes mellitus, but not to coronary heart disease, in Saudi Arabians. AB - INTRODUCTION: Plasma homocysteine (HCYS) concentration is believed to be an independent risk factor for atherosclerosis. METHODS: HCYS was measured in a cohort of 584 Saudi Arabians participating in a national screening study of coronary heart disease (CHD) risk factors. A total of 173 subjects (114 men and 59 women) had clinical CHD, of whom 82 (47.4%) had type 2 diabetes mellitus (56 men and 26 women). A further 127 subjects (60 men and 67 women) also had type 2 diabetes mellitus but no CHD. A total of 284 individuals (120 men and 164 women) were recruited as healthy controls, and had no previous history of CHD or diabetes. Serum HCYS was measured by high-performance liquid chromatography (HPLC) with electrochemical detection. RESULTS: Univariate analysis showed HCYS concentrations were significantly lower in those with diabetes mellitus (DM) than in controls, for both men [8.7 (4.2-18.6) vs. 10.5 (4.5-20.5) mmol/l, median (5th 95th percentiles, p = 0.009] and women [6.3 (3.3-24.0) vs. 8.1 (4.0-17.9) mmol/l, p = 0.049]. Stepwise multivariate regression analysis indicated a relationship between HCYS concentration and age, sex and the presence of DM, but not with CHD. CONCLUSIONS: In the Saudi Arabian population, serum HCYS is not a risk factor for CHD, but is lower in patients with DM. PMID- 11940110 TI - Effects of arachidonic acid plus zinc on glucose disposal in genetically diabetic (ob/ob) mice. AB - AIM: The present study is designed to determine whether arachidonic acid (AA) plus zinc improves clinical signs of diabetes in genetically diabetic ob/ob mice. METHODS: In the first study, effects of acute administration of AA plus zinc on glucose disposal were determined in ob/ob and lean mice (n = 6 each). In the second study, ob/ob and lean mice were treated with increasing doses of AA plus zinc for 2 weeks (n = 5 each). Postprandial and fasting blood glucose concentrations, three-hour-area-average above fasting glucose concentration (TAFGC), water and food intake, body weight and plasma insulin concentrations were measured. RESULTS: Acute administration of AA plus zinc significantly increased glucose disposal in ob/ob mice. In the second study, postprandial and fasting blood glucose concentrations, TAFGC, and water and food intake in ob/ob mice treated with AA plus zinc for 2 weeks were significantly decreased compared with those in mice given no AA. Plasma insulin concentrations in both lean and ob/ob mice were not changed by AA treatment in drinking water. CONCLUSIONS: AA plus zinc in drinking water is effective in decreasing blood glucose levels in obese mice. These results indicate that use of these compounds should be considered as a dietary supplement to control hyperglycaemia in patients with type II diabetes. PMID- 11940111 TI - Glucose screening during active labour for the detection of undiagnosed hyperglycaemia and newborn malformations in a population without prenatal care. AB - AIM: This prospective study was undertaken to look at the frequency of hyperglycaemia in pregnant women who were without health benefits and did not receive prenatal specialist care, and to examine the relationship between fetal complications and the prenatal care received. METHODS: The subjects consisted of 304 pregnant women, divided into two groups. Group I included 176 women who had inadequate prenatal care (73 women with irregular care, and 103 women without any specialist care.) with a mean of 1.1 visits to a specialist. Group II included 128 women who had regular prenatal care with a mean of 6.5 consultations per patient. RESULTS: There were no statistical differences between the two groups in relation to maternal age, obstetrical history and gestational weeks. The familial antecedent of diabetes was found in 30.1% of women in group I and 19.5% of women in group II (p < 0.05). Hyperglycaemia was found in 55 (31.2%) patients in group I and 4 (3.1%) patients in group II (p < 0.001) with mean blood glucose levels of 163 and 130 mg/dL respectively. Clinical characteristics of newborns show statistically significant increased abnormalities in 15.9% of babies whose mothers had inadequate prenatal care vs. 3.9% of babies whose mothers had regular prenatal care. The abnormalities seen were myelomeningocele, cleft lip and palate, oesophageal atresia, polydactyly, phocomelia and encephalocele. CONCLUSIONS: The frequency of hyperglycaemia in pregnant women with inadequate prenatal care was higher than in pregnant women with regular prenatal care. Additionally newborn abnormalities and complications occurred more frequently in babies of mothers with inadequate prenatal care. The benefits of prenatal care are evident and it is important to improve and start instituting new programmes and ways of making health information available to women in primary care clinics to educate the general population and stress the importance of regular visits to a prenatal care specialist. PMID- 11940112 TI - Using nuclear targeting signals to enhance non-viral gene transfer. AB - Summary Gene therapy involves the introduction of DNA-encoding therapeutic gene products into appropriate cells of an affected individual. The limitations of the approach relate largely to the poor efficiency of the delivery of the therapeutic DNA to the nucleus. This review examines recent work in the area of non-viral gene transfer, building on developments in the field of nuclear protein import and their application in the field of non-viral gene transfer. In particular, advances in the area of enhancing DNA targeting to the nucleus are discussed, including the use of modular nuclear targeting signals recognised by the cellular nuclear import machinery and DNA condensing agents to facilitate passage through the nuclear pore. Optimising nuclear DNA delivery through these and other strategies should assist greatly in rendering gene therapy a viable and realistic possibility for treating disease. PMID- 11940113 TI - Activation-induced cell death: the controversial role of Fas and Fas ligand in immune privilege and tumour counterattack. AB - Activation-induced cell death (AICD) is the process by which cells undergo apoptosis in a controlled manner through the interaction of a death factor and its receptor. Programmed cell death can be induced by a number of physiological and pathological factors including Fas (CD95)-Fas ligand (FasL/CD95L) interaction, tumour necrosis factor (TNF), ceramide, and reactive oxygen species (ROS). Fas is a 48-kDa type I transmembrane protein that belongs to the TNF/nerve growth factor receptor superfamily. FasL is a 40-kDa type II transmembrane protein that belongs to the TNF superfamily. The interaction of Fas with FasL results in a series of signal transductions which initiate apoptosis. The induction of apoptosis in this manner is termed AICD. Activation-induced cell death and Fas-FasL interactions have been shown to play significant roles in immune system homeostasis. In this review the involvement of Fas and Fas ligand in cell death, with particular reference to the T cell, and the mechanism(s) by which they induce cell death is described. The role of AICD in immune system homeostasis and the controversy surrounding the role of FasL in immune privilege, inflammation, and so-called tumour counterattack is also discussed. PMID- 11940114 TI - Interleukin-dependent modulation of HLA-DR expression on CD4and CD8 activated T cells. AB - Summary Interleukins (IL) regulate different T-cell surface Ag known as activation markers that have distinct functional roles. In this paper, while studying the influence of some cytokines(IL-12, IL-2 and IL-4) on the expression of several markers [CD69,CD25, CD26, CD3, human leukocyte antigen (HLA-DR), CD45R0] in in vitro activated human T lymphocytes, we observed two groups of donors responding to phytohaemagglutinin (PHA) activation with high or low HLA DRAg expression. We also found that CD4 and CD8 populations had different HLA-DR densities under PHA activation (particularly the high HLA-DR-expressing group). Interleukins, in a dose-dependent manner (IL-2 partially),upregulated these HLA DR levels. In 5 day cultures, IL-12 and IL-2 enhanced the CD8/CD4 ratio of activated T cells,which was responsible, in part, for the IL-dependent HLA-DR upregulation.IL-12 and IL-2 also upregulated the HLA-DR expression at the molecular level on CD8, and IL-12 downregulated it on CD4 cells. It seems that IL 4 upregulated HLA-DR by shortening the mitogen-dependent regulation kinetics. We hypothesize that the different effect of each IL on HLA-DR expression might be related to the regulation of the dose of antigenic peptide presentation and, thus, also influence TH1/TH2 dominance. PMID- 11940115 TI - Temporal changes in the distribution of thoracic duct lymphoblasts to synovium and other tissues of rats with adjuvant-induced arthritis. AB - The distribution of lymphoblasts(lymphocytes in cell cycle) obtained from the central lymph of donor rats and transferred adoptively to syngeneic recipients has been shown previously to be influenced by the presence of arthritis in either donor or recipient rats. The intent of the present study was to examine patterns of distribution of lymphoblasts in the early period after transfer, when extravasation of donor lymphoblasts was expected to occur. Thoracic duct lymphoblasts labelled in vitro with [125I]-iododeoxyuridine were detected in recipient rats by external radiometry and autoradiography. Irrespective of donor status, fewer donor lymphoblasts accumulated in the feet of normal recipients when compared to arthritic recipients at 15 min, 2 h and 24 h after cell transfer.When recipients of similar disease status were compared, the percentages of injected lymphoblasts from normal and arthritic donors recovered in the feet were similar at 15 min and 2 h after transfer. The proportions of lymphoblasts recovered in the feetat 24 h after injection declined in normal recipients and arthritic recipients of cells from normal donor rats. Importantly,this decline did not occur when both the donor and the recipient were arthritic. In the hindpaws, donor lymphoblasts were located predominantly in the bone marrow, except in transfers between arthriticrats, when at 24 h they were predominantly in the synovium. At 15 min, lymphoblasts were detected within the lumen of vessels within synovium, whereas by 2 h extravasation of these cells was evident. In conclusion, lymphoblasts accumulate more readily in hindfeet that are inflamed. In the early hours after injection, lymphoblasts from normal and arthritic donors are recruited equally, but these early levels are only maintained for 24 hin the combination of arthritic donor and arthritic recipient. Adramatic change in the proportion of lymphoblasts located in synoviumat this later time suggests that a dynamic process of relocation,retention and/or local cell division maintains the numbers of arthritic donor cells in the latter combination. PMID- 11940116 TI - Characterization of two TCR transgenic mouse lines specific for herpes simplex virus. AB - To better understand the T cell-mediated processes involved in the immune response to herpes simplex virus type 1 (HSV-1)infection, two HSV-specific T cell receptor (TCR) transgenic mouse lines were produced. These mice (gBT-I.1 and gBT I.3) are MHC class I-restricted and specific for the immunodominant peptide from HSV glycoprotein B (gB), gB498-505. Although derived from the same clone, the mice differ in the chromosomal location of the TCR transgenes and show marked differences in TCR alpha/beta expression on both CD4+ and CD8+ cells in the thymus. Despite this, peripheral CD8+ Tcells from both mice express equally high levels of the transgenic TCR and bind the KbgB498-505 tetramer to the same degree. In concordance with this, both were shown to respond equally well in vitro upon stimulation with the gB498-505 peptide or HSV-infected cells. These data show that selection of broadly equivalent peripheral T-cell subsets can occur in the presence of distinctly different thymic T-cell subsets. PMID- 11940117 TI - Nuclear RelB+ cells are found in normal lymphoid organs and in peripheral tissue in the context of inflammation, but not under normal resting conditions. AB - Differentiated dendritic cells (DC) have been identified by the presence of nuclear RelB (nRelB) and HLA-DR, and the absence of CD20 or high levels of CD68, in lymph nodes and active rheumatoid arthritis synovial tissue. The current studies aimed to identify conditions in which nRelB is expressed in human tissues, by single and double immunohistochemistry of formalin-fixed peripheral and lymphoid tissue. Normal peripheral tissue did not contain nRelB+ cells. nRelB+ DC were located only in T- or B-cell areas of lymphoid tissue associated with normal organs or peripheral tissues, including tonsil, colon, spleen and thymus, or in association with T cells in inflamed peripheral tissue. Inflamed sites included skin delayed-type hypersensitivity reaction, and a wide range of tissues affected by autoimmune disease. Nuclear RelB+-HLA-DR- follicular DC were located in B-cell follicles in lymphoid organs and in lymphoid-like follicles of some tissues affected by autoimmune disease. Lymphoid tissue T-cell areas also contained nRelB(-)-HLA-DR+ cells,some of which expressed CD123 and/or CD68. Nuclear RelB+ cells are found in normal lymphoid organs and in peripheral tissue in the context of inflammation, but not under normal resting conditions. PMID- 11940118 TI - Mast cells induce upregulation of P-selectin and intercellular adhesion molecule 1 on carotid endothelial cells in a new in vitro model of mast cell to endothelial cell communication. AB - It is suggested that mast cells contribute to cell recruitment in inflammation through the upregulation of endothelial adhesion molecules. P-selectin and intercellular adhesion molecule(ICAM)-1 are two key adhesion molecules that have been associated indirectly with mast cell activity. The canine C2 mastocytoma cell line and primary cultures of canine carotid endothelial cells were used to establish a new in vitro model to help study the interaction between mast cells and endothelial cells. Carotid endothelial cells were incubated with mast cell mediators to uncover their effect on endothelial ICAM-1 and P-selectin expression. To assess the relative contributions of tumour necrosis factor (TNF) alpha and histamine to such effect, an H1 antihistamine and a TNF-alpha blocking antibody were used. Prior to activation by mast cell mediators, P-selectin was expressed only within the cytoplasm, and ICAM-1 was constitutively expressed on the surface of the canine carotid endothelial cells. Both adhesion molecules were enhanced significantly and strongly upon mast cell activation at various time points. Unstored TNF-alpha was fully responsible for ICAM-1 upregulation. P selectin was up-regulated by both preformed and newly synthesized mast cell mediators, but neither histamine nor TNF-alpha accounted for such an effect. Therefore,a new model is proposed in which the pro-inflammatory effect of mast cells on endothelial cells can be studied in vitro. In this model, it has been demonstrated that only TNF-alpha accounts for the overexpression of ICAM-1 induced by mast cells, and that mast cells up-regulate P-selectin expression through a histamine-independent mechanism. PMID- 11940120 TI - Macrophages and nitric oxide as the possible cellular and molecular basis for strain and gender differences in susceptibility to autoimmune central nervous system inflammation. AB - PVG rats are resistant to actively induced experimental autoimmune encephalomyelitis (EAE) and this appears to be directly related to high and sustained systemic levels of reactive nitrogen intermediates(RNI) following sensitization for EAE when compared to the highly susceptible Lewis rat. An apparent cellular basis for the different EAE susceptibility between the two rat strains is described. Spleens of PVG rats have increased monocyte/macrophage numbers(NO producing cells) and lower erythrocyte (NO scavengers) to nucleated spleen cell ratios compared with Lewis rats. Splenectomy demonstrated the pivotal role of the spleen in resistance to EAE as splenectomized PVG rats were rendered completely susceptible to disease induction. It was further demonstrated that EAE resistance in PVG rats is limited only to females and that only female PVG rats have increased splenic macrophage and an enhanced NO production following immunization. The males are fully susceptible to EAE and their spleen cell populations are similar to those of Lewis rats of either gender. Despite being resistant to active disease induction, immunized female PVG rats can generate EAE effector cells that are capable of passively transferring disease.Furthermore, female PVG rats are fully susceptible to passively transferred EAE. Thus, there appears to be no defect in the female PVG target tissue or in the processing or presentation of antigen,but a block at the level of effector cell expansion and/or recirculation and transmigration into the target tissue in actively induced EAE. PMID- 11940119 TI - Enhancing the immunogenicity and modulating the fine epitope recognition of antisera to a helical group A streptococcal peptide vaccine candidate from the M protein using lipid-core peptide technology. AB - A conserved helical peptide vaccine candidate from the M protein of group A streptococci, p145, has been described. Minimal epitopes within p145 have been defined and an epitope recognized by protective antibodies, but not by autoreactive T cells, has been identified. When administered to mice, p145 has low immunogenicity. Many boosts of peptide are required to achieve a high antibody titre (> 12 800). To attempt to overcome this low immunogenicity, lipid core peptide technology was employed. Lipid-core peptides (LCP) consist of an oligomeric polylysine core, with multiple copies of the peptide of choice, conjugated to a series of lipoamino acids, which acts as an anchor for the antigen. Seven different LCP constructs based on the p145 peptide sequence were synthesized (LCP1-->LCP7) and the immunogenicity of the compounds examined. The most immunogenic constructs contained the longest alkyl side-chains. The number of lipoamino acids in the constructs affected the immunogenicity and spacing between the alkyl side-chains increased immunogenicity. An increase in immunogenicity (enzyme-linked immunosorbent assay (ELISA) titres) of up to 100 fold was demonstrated using this technology and some constructs without adjuvant were more immunogenic than p145 administered with complete Freund's adjuvant (CFA). The fine specificity of the induced antibody response differed for the different constructs but one construct, LCP4, induced antibodies of identical fine specificity to those found in endemic human serum. Opsonic activity of LCP4 antisera was more than double that of p145 antisera. These data show the potential for LCP technology to both enhance immunogenicity of complex peptides and to focus the immune response towards or away from critical epitopes. PMID- 11940121 TI - Intrinsic in vitro abnormalities in dendritic cell generation caused by non-MHC non-obese diabetic genes. AB - Genes outside the MHC create a general susceptibility to autoimmunity in non obese diabetic (NOD) mice. In this study, we describe marked differences in dendritic cell generation, in vitro, caused by non-MHC NOD genes. Bone marrow cells from NOD.H-2k mice cultured in vitro with GM-CSF and IL-4 generated a reduced yield of dendritic cells when compared to bone marrow cells from B10.H-2k mice. This was due to failure to pass through successive rounds of cell division and elevated levels of apoptosis in NOD.H-2k precursor cells. This aberrant response to GM-CSF and IL-4 was unique to the NOD.H-2k background when compared to bone marrow cells from other H-2k congenic strains, and coculture experiments showed that it was cell-autonomous. Overall, the results described in this study demonstrate a striking effect of non-MHC NOD genes on dendritic cell generation from myeloid precursors derived from the NOD.H-2k strain. These results identify a useful genetic model to explore the regulation of dendritic cell formation. Conceivably, the dysregulation of the dendritic cell system described here may contribute to the generalized defects in self-tolerance in the NOD strain. PMID- 11940122 TI - Nursing is essential to improving patient safety. PMID- 11940124 TI - Analysis of the relationship between nurse influences over flexible working and commitment to nursing. AB - BACKGROUND: The paper explores the theoretical and practical bases of both commitment and control within the context of temporal aspects of flexible working in nursing. AIM: The aims of the paper are to examine the relationships between nurses' shift patterns, influence over shift pattern and realization of shift preference and commitment to nursing. METHODS: Data were collected through a postal questionnaire completed by 2987 British nurses employed in hospitals, care homes and hospices. Principal components analysis was used to identify common factors among responses to a series of 33 statements about working life. Data were analysed using ANOVA and multiple regression techniques. RESULTS: Permanent night shift nurses reported lower levels of commitment to nursing. As predicted, influence over shift patterns and realization of preferred shift pattern were positively associated with commitment to nursing, although the relationship was weak. Positive perceptions of career development opportunities were a stronger predictor of commitment to nursing. Results are discussed in light of previous ethnographic research on nurses' shift patterns. CONCLUSION: The opportunity to explore quantitatively the effects of shift-related decisions on commitment using a large sample is useful. While influence and shift type were predictors of commitment, the positive impact they were expected to have was smaller than expected. Similarly, the negative effects of not having influence or of working permanent night shifts were smaller than expected and the statistical significance of such small effects relies heavily on the large sample obtained. Other variables, particularly career development prospects, outweigh the influence of shift-related variables on commitment. The research gives a clear message to human resource managers involved with nurse management: so long as nurses have a strong perception of career development potential the otherwise negative influences of shift impact can be minimized. PMID- 11940125 TI - Online learning: enhancing nurse education? AB - BACKGROUND: The need to integrate information technology into nursing education has been recognized and well documented. In spite of this, information technology remains a neglected subject in many nursing programmes. Strategies have been considered for increasing the integration of information technology in nursing education. One of the key issues identified is the need for research into the factors that contribute to optimal learning with information technology, specifically the need to explore issues that contribute to student frustration and satisfaction with learning. Within Australia, the incorporation of information technology as a core subject in nursing education is still relatively new. This article describes how one university used 'online' learning to expose students to conceptual and experiential opportunities that enabled them to develop skills in the management of information technology. METHODS: Twenty-one students participated in this qualitative study. Individual interviews were used to develop insights into student perceptions. Thematic analysis enabled refined themes to emerge. These themes formed the basis of focus group discussions. Focus groups were used to enhance and validate the information from one-to-one interviews by using group dynamics to add experiential richness to the data. FINDINGS: Four major themes emerged: computer confidence, flexibility, active learning and practicalities of teaching. CONCLUSIONS: The integration of information technology into nursing education requires a dramatic change in thinking. The 'learning curve' is steep for both student and educator and there are many issues that need to be considered. This research does not aim to provide solutions to the issues highlighted but rather offers recommendations for enhancing the teaching and learning experience. PMID- 11940126 TI - Unravelling the unknowns of reflection in classroom teaching. AB - BACKGROUND: The use of reflection in education has emerged as an effective means of connecting theory with practice. However, the literature reveals limited empirical work on the conceptualization of reflection. AIM OF THE STUDY: The purpose of this qualitative research study was to understand more fully the meaning and use of reflection in teaching, and how reflection contributes to the development of teaching expertise in the classroom. Ethical approval. The study received ethical approval from the Faculty of Nursing Ethical Review Committee, University of Manitoba. Issues related to confidentiality of information, and power relationships between the investigators were addressed. Students were reassured that no data were collected about them during participant observations in the classrooms. METHODS: The investigators were the sources of data. Data were collected using five different methods over the course of two academic years, including written autobiographies, critical incident journals, classroom observations, debriefing following classroom observations and research team meetings. FINDINGS: The data were analysed using content analysis, and four themes were identified (i) making connections, (ii) developmental aspects, (iii) influence of context on reflection, and (iv) influence of emotions on reflection. Study limitations. The interpretation of the findings of this study should be used with caution given the qualitative design and small number of participants. CONCLUSION: Participation in the study increased the awareness of the investigators' personal use of reflection. The process of studying our own use of reflection allowed us to step outside the performance treadmill to better understand, accept and reshape what we do over and over in the classroom. This study supports an examination of one's experiences as a means of understanding reflection and its use in the classroom. PMID- 11940127 TI - Rituals and nursing: a critical commentary. AB - AIM OF THE PAPER: This critical commentary explores the concept of ritual in the nursing and social science literature in terms of investigating different understandings of its meaning and purpose. RATIONALE: The term 'ritual' is often used in a pejorative sense in nursing literature to refer to unthinking, routinized action by nurses, which lacks any empirical foundation. The purpose of this paper is to explore alternative understandings of the meaning of ritual, suggesting that the negative usage described above misses out on the wider symbolic meaning of the word. METHOD: The literature search used a combination of electronic databases (CINAHL, ASSIA, MEDLINE and RCN) from 1960 to 2000 and serendipitous references in texts. FINDINGS: The commentary is presented under the following two broad emerging themes: first, the complex issues surrounding the definition of ritual, including the relationship between rational and rational action and second, the purposes served by ritual. CONCLUSION: The paper concludes with the recommendation that the rituals nurses use in the performance of their care are worthy of research as a rich source of insight into the meaning and purpose of nursing actions. PMID- 11940128 TI - Impact of a nurse-led counselling service on quality of life in patients with inflammatory bowel disease. AB - AIM: Health related quality of life is impaired in patients suffering from inflammatory bowel disease. Although counselling directed towards physical and psychological morbidity is assumed to improve health related quality of life, this has never been demonstrated. METHODS: Physical and psychological well-being were assessed using questionnaires administered to 100 out-patients in the United Kingdom suffering from inflammatory bowel disease, 50 subjects not suffering from inflammatory bowel disease and a disease control group comprising 28 patients with psoriatic arthritis. A specific nurse led counselling package was given to half the inflammatory bowel disease group and health related quality of life was assessed at baseline, 6 and 12 months. RESULTS: Inflammatory bowel disease and psoriatic arthritic patients had a range of physical disease activity, although none were severely ill during the course of the study. Medical therapy was similar in both groups throughout the duration of the trial. The mean Short Form 36 (SF-36) scores for mental health were low in inflammatory bowel disease patients; 62.9 +/- 9.1 (SD) in ulcerative colitis, 60 +/- 9.8 (SD) in Crohn's disease, compared with 72.4 +/- 7.2 (SD) in healthy controls (P < 0.05). Mean SF 36 scores for social function were also reduced in Crohn's disease patients; 68.4 +/- 10.1 (SD) in Crohn's disease, compared with 87 +/- 10.1 (SD) in healthy controls (P < 0.05). As expected, the mean SF-36 scores in psoriatic arthritic patients were significantly low 61.9 +/- 1.5 (SD) compared with 82.4 +/- 14 (SD) in healthy controls (P < 0.05). Crohn's disease patients were significantly more anxious than the other groups, mean HAD score was 10 +/- 3.7 (SD) in Crohn's disease patients and 6.86 +/- 3.5 (SD) in healthy volunteers (P < 0.05), although mean HAD scores for depression were similar in all groups. Maladaptive coping mechanisms were present in a significant proportion of Crohn's disease patients. At follow-up all aspects of psychological morbidity returned to the normal range in the Crohn's disease patients without significant change in the mean physical disease index. CONCLUSION: Health related quality of life can be improved over 6 months by provision of a nurse led counselling service but the effects are not sustained for 12 months. PMID- 11940129 TI - Clinical innovation for promoting family care in paediatric intensive care: demonstration, role modelling and reflective practice. AB - AIM: To explore family caregiving problems in paediatric crisis care and methods that could be applied to move the abstraction of family care to development of specific family interventions. BACKGROUND AND RATIONAL: Family centred care has been accepted as the ideal philosophy for holistic health care of children, but methods for its implementation are not well established. In paediatric health crises, family care requires special sensitivity to family needs and a type of complex nursing care for which many practitioners are not sufficiently prepared. Developing family sensitive models of intervention and finding a strategy for transfer of this knowledge to clinical practice is an important challenge facing family nursing today. Social learning theory provides a rich background to explore these issues. CONCLUSIONS: Specific techniques of role modelling and reflective practice are suggested as effective approaches to teach family sensitive care in clinical settings where families are part of the care environment. PMID- 11940130 TI - Research and organizational issues for the implementation of family work in community psychiatric services. AB - BACKGROUND: The need for evidence-based practice (EBP) to guide and develop mental health services remains fundamental for modern services. Aim. To discuss issues that impact upon implementation of EBP and practice development using family work (FW) as an example. METHODS: A selection of the FW literature was reviewed drawing on sources including the Cochrane Library, Cinahl and Medline. Keywords used were FW, community mental health team and research design. FINDINGS: Centralized policy initiatives and guidelines that are themselves guided by evidence of randomized controlled trials predominantly risk alienating practitioners and clients/carers. Family work has some demonstrable clinical benefits although models differ and the active therapeutic agent remains unclear. Its adoption into routine care is also hindered by a productivity management outlook that seeks to maximize stretched resources and whose values are likely to be internalized by practitioners. The dichotomous position of previous research and practice development make implementation of EBP difficult and highlights the need for strategic planning that embraces both factors. CONCLUSION: The current drive to increase EBP requires a bi-directional process of influence that allows individual practitioners and clients/carers to become producers of evidence and not simply recipients. The authors support wider adoption of case study research designs to reflect the unpredictable nature of mental health care. Adoption of assertive community treatment models within community services is most likely to promote the excellence management model and accommodate EBP such as FW. PMID- 11940131 TI - Cuff width influence on blood pressure measurement during the pregnant-puerperal cycle. AB - BACKGROUND: The correct evaluation of blood pressure (BP) during pregnancy is a crucial factor in the prevention of eclampsia. Following American Heart Association (AHA) guidelines to employ a correct cuff width (CCW), 20% larger than arm diameter, we demonstrate that the standard cuff width (SCW), 12 cm wide, is too large for lean women causing underestimation of BP. AIMS: To identify the arm circumference (AC) in pregnant women and the corresponding cuff width; to compare BP records from CCW vs. SCW; and to identify under- and overestimation of BP in SCW reading. DESIGN: A follow up study of BP was performed in 104 pregnant women using two cuffs widths (CCW vs. standard one). The investigation was carried out during all antenatal appointments and postpartum stage in two maternity hospitals. In every appointment BP was registered three times with each type of cuff; and the means of those three readings were compared. METHODS: The CCW for each woman was selected according to AHA recommendation for cuff width size (20% larger than arm diameter), which was based on the classical European and North-American studies. Results. Arm circumference varied from 20 to 38 cm requiring a cuff width from 8 to 14 cm. The CCW most employed was 10 cm wide. The SCW (12 cm) was employed as CCW in only 13.4% of the subjects. Statistical difference was found on BP means when comparing both cuffs (P < 0.05), reaching 23 mmHg in systolic values and 20 in diastolic ones. Such differences showed a serious underestimation when SCW was employed in 80.8% of the subjects and overestimation in 5.8% of obese subjects. CONCLUSION: Our findings showed that the SCW underestimates BP of pregnant women. Our hypothesis is that such underestimation may lead to the misdiagnosis of pre-eclampsia, particularly in lean pregnant women. PMID- 11940132 TI - A risk assessment scale for the prediction of pressure sore development: reliability and validity. AB - BACKGROUND: The ability to assess the risk of a patient developing pressure sores is a major issue in pressure sore prevention. Risk assessment scales should be valid, reliable and easy to use in clinical practice. AIM: To develop further a risk assessment scale, for predicting pressure sore development and, in addition, to present the validity and reliability of this scale. METHODS: The risk assessment pressure sore (RAPS) scale, includes 12 variables, five from the re modified Norton scale, three from the Braden scale and three from other research results. Five hundred and thirty patients without pressure sores on admission were included in the study and assessed over a maximum period of 12 weeks. Internal consistency was examined by item analysis and equivalence by interrater reliability. To estimate equivalence, 10 pairs of nurses assessed a total of 116 patients. The underlying dimensions of the scale were examined by factor analysis. The predictive validity was examined by determination of sensitivity, specificity and predictive value. RESULTS: Two variables were excluded as a result of low item-item and item-total correlations. The average percentage of agreement and the intraclass correlation between raters were 70% and 0.83, respectively. The factor analysis gave three factors, with a total variance explained of 65.1%. Sensitivity, specificity and predictive value were high among patients at medical and infection wards. CONCLUSION: The RAPS scale is a reliable scale for predicting pressure sore development. The validity is especially good for patients undergoing treatment in medical wards and wards for infectious diseases. This indicates that the RAPS scale may be useful in clinical practice for these groups of patients. For patients undergoing surgical treatment, further analysis will be performed. PMID- 11940133 TI - Using the Zelen design in randomized controlled trials: debates and controversies. AB - BACKGROUND: The use of the randomized consent design (commonly known as the Zelen design) is a controversial issue in randomized controlled trials. In the Zelen design, participants are randomly allocated prior to seeking consent. Those participants allocated to the intervention group are then approached and offered the intervention, which they can decline or accept. Zelen first proposed the design in 1979. It has been used infrequently since this time, although there are some notable exceptions in nursing, midwifery and some medical specialities. AIM: This paper describes the Zelen design, including the two forms used (the single and double consent versions) and discusses the advantages and disadvantages of using such a design. METHODS: An explanation of the differences between the Zelen design and a conventional randomized controlled trial is presented. In a conventional design, detailed knowledge of the alternative interventions is given to the prospective participant. The participant gives consent and is allocated to one of the groups. In a Zelen design, participants are randomly allocated and then approached and offered the group to which they were allocated. The Zelen design is used firstly, to reduce disappointment bias in the conventional consent randomization process, and secondly, to remove subjective bias in the recruitment process. There are concerns relating to the use of the Zelen design, including ethical concerns relating to the timing of random allocation and consent and the collection of clinical data. CONCLUSION: It is hoped that by presenting issues pertaining to the Zelen design, other nursing and midwifery researchers may be prompted to consider its use when designing clinical research. The Zelen design is controversial, and debate about its merits and shortcomings is useful. This paper contributes to the ongoing debate. PMID- 11940135 TI - The antioxidant capacity of saliva. AB - BACKGROUND/AIMS: Saliva, a heterogeneous fluid comprising proteins, glycoproteins, electrolytes, small organic molecules and compounds transported from the blood, constantly bathes the teeth and oral mucosa. It acts as a cleansing solution, an ion reservoir, a lubricant and a buffer. In addition to its other host-protective properties, saliva could constitute a first line of defence against free radical-mediated oxidative stress, since the process of mastication and digestion of ingested foods promotes a variety of reactions, including lipid peroxidation. Moreover, during gingival inflammation, gingival crevicular fluid flow increases the change of saliva composition with products from the inflammatory response; this, in turn, could have some role in controlling and/or modulating oxidative damages in the oral cavity. This is the reason why the antioxidant capacity of saliva has led to increasing interest, and the development of techniques suitable for saliva antioxidant evaluation. MATERIALS AND METHODS: Here, we review the current peer-reviewed literature concerning the nature and characteristics of free radicals, reactive oxygen species, oxidants, pro-oxidants and antioxidants in saliva, especially pro oxidant and antioxidant features, as well as current methods for assessing the antioxidant capacity of saliva. RESULTS AND CONCLUSIONS: In the last decade, several methods have been developed for assaying the antioxidant activity of saliva, indicating an increasing interest of researchers and clinicians. Unfortunately, systematic studies of saliva are still lacking, even in healthy populations. PMID- 11940136 TI - Comparison of the bleeding on marginal probing index and the Eastman interdental bleeding index as indicators of gingivitis. AB - AIM: The purpose of the present study was to compare 2 indices, i.e., the Eastman interdental bleeding (EIB) index and the bleeding on marginal probing (BOMP) index. The comparison was made (a) in terms of the degree of bleeding provoked and the relationship with plaque in natural gingivitis and (b) for the ability of these 2 methods to detect differences between the development of experimental gingivitis in a control group and a group in which the development of gingival inflammation was suppressed by treatment. For the present studies, subjects were selected without interdental recession of the gingival tissues. METHODS EXPERIMENT 1: In this experiment, 43 subjects having established moderate gingivitis were assessed using a random splitmouth design (1st and 3rd/2nd and 4th quadrant). Plaque was scored on all approximal sites after which the BOMP index was assessed in one half of the mouth and the EIB index in the other. RESULTS EXPERIMENT 1: The BOMP index showed a bleeding score of 84% and the EIB index of 87%. The significant correlation between plaque and gingival bleeding for the BOMP index (0.55) was higher than for the EIB index (0.44). METHODS EXPERIMENT 2: For this experiment, 25 subjects participated in an experimental gingivitis trial of the lower jaw. At baseline, first the BOMP index and immediately thereafter the EIB index were assessed at all approximal sites. Experimental gingivitis (EG) was carried out in one randomly assigned quadrant and as a treatment modality only floss was used in the other (FL). RESULTS EXPERIMENT 2: In the EG quadrant, the BOMP index increased to 69% and the EIB index to 73%. Both indices showed a significant correlation with plaque; 0.60 and 0.64 respectively. In the FL quadrant, the BOMP index increased to 38% and the EIB index to 30%. No significant correlation between both gingivitis indices and the amount of plaque was present in the FL quadrant. CONCLUSION: The ability of the BOMP index and the EIB index to assess the level gingival inflammation appears to be comparable. PMID- 11940137 TI - White blood cell count in generalized aggressive periodontitis after non-surgical therapy. AB - BACKGROUND: Periodontal bacteria are known to invade the systemic circulation. Chronic low-level bacteremia and a systemic inflammatory response have been suggested as a pathogenetic link between periodontal disease and atherosclerosis. The purpose of this study was to examine the systemic effect of a non-surgical therapy on white blood cell count (WBC count) and differential blood count in smoking and non-smoking generalized aggressive periodontitis (GAP) patients. METHODS: 27 adult periodontitis patients (13 smokers and 14 non-smokers) with previously untreated GAP were subjected to 3 sessions of oral hygiene procedure. Afterwards, the patients were treated by scaling and root planing under local anaesthesia. Periodontal examinations were performed after supragingival pretreatment and three months after subgingival therapy. Pocket probing depth (PPD) and relative attachment level (RAL) were measured with Florida probe and disc probe. Accompanying clinical evaluation venous blood samples were taken to analyse the WBC counts and differential blood counts. For statistical analysis non-parametric tests were utilized. RESULTS: No clinical or demographic differences were found between smokers (n=13) and non-smokers (n=14). PPD, bleeding on probing (BoP) and suppuration improved significantly after therapy both in smokers and non-smokers. Following periodontal treatment WBC counts, neutrophil and platelet counts decreased significantly in non-smokers (p< or =0.004), while in smokers only platelet counts were significantly reduced (p=0.006). Non-smokers showed a significantly higher reduction of WBC counts (p=0.005) and neutrophils (p=0.001) compared to smokers. CONCLUSION: The results indicate that a therapeutical intervention may have a systemic effect on the blood count in GAP patients. This effect seems to differ between smokers and non smokers. PMID- 11940138 TI - Anti-plaque effect of tempered 0.2% chlorhexidine rinse: an in vivo study. AB - OBJECTIVES: The aim of this in vivo study was to compare the anti-plaque effect of warm and cold chlorhexidine gluconate irrigation on matured human plaque. METHODS: In a split-mouth design, the antibacterial effect of 47 degrees C "warm" 0.2% chlorhexidine solution was compared with that of 18 degrees C "cold" rinse at the same concentration on newly-formed supragingival plaque that had been left undisturbed for 72 h. Before and 1 h after a 1-min rinse procedure, plaque was sampled from 10 test persons and vitality determined using vital fluorescence technique. RESULTS: Cold and warm 0.2% chlorhexidine solution reduced plaque vitality significantly from 99.63% to 77.81% (p=0.014) and from 98.98% to 51.77% (p<0.001), respectively. Rinsing with warm chlorhexidine solution reduced plaque vitality to a significantly greater degree (p=0.003) than did cold chlorhexidine. CONCLUSION: In this study, warm 0.2% chlorhexidine rinse showed a significantly more intensive anti-plaque effect than cold chlorhexidine solution at the same concentration. PMID- 11940139 TI - Desensitizing effects of an Er:YAG laser on hypersensitive dentine. AB - AIM: The aim of the present study was to evaluate and compare the desensitizing effects of an Er:YAG laser (KEY II(R), KaVo, Germany) and Dentin Protector (Vivadent, Germany) on cervically exposed hypersensitive dentine. METHOD: A group of 30 patients showing a total of 104 contralateral pairs of hypersensitive and caries-free teeth was selected and randomly allocated in a split-mouth design to either (1) Er:YAG laser (80 mJ/pulse, 3 Hz), or (2) the application of Dentin Protector (polyurethane-isocyanate 22.5%; methylenechloride 77.5%) whereat one pair served as an untreated control in each patient. The degree of sensitivity to a thermal stimulus was determined qualitatively with an evaporative stimulus defined as a 3-s air blast at a distance of 2 mm from each site to be tested. A qualitative registration of the degree of discomfort was determined according to an arbitrary pain scale in 4 degrees. Recordings were assessed before treatment, immediately after, 1 week, 2 and 6 months after treatment by 1 blinded examiner. RESULTS: Both treatment forms resulted in significant improvements of discomfort immediately after and 1 week post treatment. After 2 months, the discomfort in the Dentin Protector(R) group increased up to 65% of the baseline score and even up to 90% after 6 months, whereas the effect of the laser remained at the same level that was achieved immediately after treatment. The differences immediately after, 1 week, 2 and 6 months post treatment between both groups were statistically high significant (p< or =0.001; respectively). Compared to the untreated control group, both treatment forms resulted in a significant reduction of discomfort at each follow-up examination. CONCLUSION: It was concluded that desensitizing of hypersensitive dentine with an Er:YAG laser is effective and the maintenance of the positive result was more prolonged than with Dentin Protector. PMID- 11940140 TI - In-dental-office screening for diabetes mellitus using gingival crevicular blood. AB - BACKGROUND: Diabetes mellitus (DM) is undiagnosed in approximately 1/2 of the patients actually suffering from the disease. In addition, the prevalence of DM is more than 2x as high in patients with periodontitis when compared to periodontally healthy subjects. Thus, a high number of patients with periodontitis may have undiagnosed DM. AIM: The purpose of this pilot study was to evaluate, whether blood oozing from gingival tissues during routine periodontal examination can be used for determining glucose levels. 32 non diabetic and 13 diabetic patients with moderate to severe periodontitis were enrolled and subjected to routine clinical periodontal examination. Periodontal pocket probing was performed using a standard force. Blood oozing from gingival tissues of anterior teeth following periodontal pocket probing was collected with the stick of a glucose self-monitoring device (Elite(R) 2000, Bayer Diagnostics GmbH, Munich). As control, fingerstick capillary blood was taken. Statistical analysis was performed by Pearson's correlation coefficient. RESULTS: The patient blood glucose levels ranged from 3.57 mmol/l to 18.01 mmol/l and the values of blood samples taken from gingiva or finger tip showed a very high intrapatient correlation (r=0.98; p<0.0001). CONCLUSION: The results suggested that blood oozing during routine periodontal examination may be used for diabetes mellitus screening in a dental office setting. PMID- 11940141 TI - How do data from deepest pocket per quadrant relate to full-mouth scores? Progression of untreated periodontal disease in young Indonesians. AB - BACKGROUND: When the subgingival presence of periodontal pathogens is studied in groups of patients or populations, mostly a number of the deepest sites is sampled. The mean clinical parameters of these deep sites are also frequently used as a the descriptor of the clinical situation of these subjects. It can be questioned, whether these 4 deep sites are capable of predicting a full-mouth situation. AIM: The purpose of the present retrospective study was to investigate to what extent the experienced progression of periodontitis as measured in the deepest approximal pocket in each quadrant reflects the disease progression at the approximal sites on a full-mouth level. METHODS: A data set of a 7-year longitudinal study of 158 young subjects (69 male, 89 female, 15-25 years of age at baseline) was used. Clinical assessments included plaque index (PI), pocket depth (PD) and attachment loss (AL) at baseline (1987) and follow-up (1994). Measurements were made at the approximal surfaces of all teeth. The deepest pocket in each quadrant was determined at follow-up. Changes of the clinical parameters between baseline and follow-up were calculated both as full-mouth mean scores as well as for these 4 deepest sites. A regression analysis was used to evaluate the relationship between full-mouth score and the 4 test sites. RESULTS: For disease progression between baseline and follow-up, significant correlation coefficients were observed between the 4-site and full-mouth mean changes (PD: 0.80, AL: 0.70, PI: 0.77). Regression coefficients were 0.51 for PD, 0.35 for AL and 0.55 for PI. The precision of the estimate for the full-mouth mean, as predicted by the 4-site mean, is determined by the residual standard deviation. This was for PD 0.31 mm, for AL 0.31 mm and for PI 0.29. Compared to the between patient standard deviation of the full-mouth means, the residual standard deviations were high. CONCLUSION: In the present population, a reasonable to good correlation between full-mouth and 4-sites data was observed. However, the high residual standard deviation in the regression analysis illustrates the inaccuracy for the 4-sites data when used as a descriptive for changes in the periodontal condition on a full-mouth level. Data evaluating progression of periodontitis based on a limited number of diseased sites should be interpreted cautiously. PMID- 11940142 TI - Collagenase-2 (MMP-8) and collagenase-3 (MMP-13) in adult periodontitis: molecular forms and levels in gingival crevicular fluid and immunolocalisation in gingival tissue. AB - AIM: To determine the cellular and molecular forms of MMP-8 (collagenase-2) and MMP-13 (collagenase-3) associated with chronic adult periodontitis by examining the species present in gingival crevicular fluid (GCF) and enzyme distribution in gingival tissue. METHODS: 30-s GCF samples were collected directly from the periodontal pockets of 12 untreated patients using filter paper strips. After elution into buffer, the samples were examined by Western immunoblotting with polyclonal antibodies for MMP-8 and MMP-13 and quantification by scanning image analysis. Individual band intensities were expressed as a percentage of total sample absorbance and mean patient values were calculated. Gingival tissue from 6 patients was fixed in formalin and embedded in paraffin wax. MMP-8 and MMP-13 were localised using the same antibodies and an avidin-biotin-peroxidase detecting system. Double staining was performed with a contrasting substrate reaction. RESULTS: The majority of MMP-8 staining in pre-treatment GCF was present in 80, 75 and 60 kD bands corresponding to prepro-, pro- and active forms of PMN-type enzyme. 43 and 38 kD bands evidently represented active, fibroblast type MMP-8. Immunoreactivities at >100 kD and < or =30 kD were probably enzyme inhibitor complex and degraded fragments, respectively. MMP-13 was seen mainly as 60 kD proenzyme with some 40 kD active enzyme and a small proportion of >100 kD complex. The percentages of MMP-8 PMN-type enzyme and MMP-13 proenzyme bands correlated significantly with gingival and bleeding indices (p<0.05). Immunohistochemistry demonstrated MMP-8 in PMNs, sulcular epithelial and also plasma cells in inflamed gingival connective tissue. MMP-13 immunoreactivity was detected in the sulcular epithelium and in macrophage-like cells. CONCLUSION: Multiple species and elevated levels of both MMP-8 and MMP-13 from many rather than single cellular sources in the diseased periodontium are identified in untreated periodontitis GCF and active forms contribute to GCF collagenase activity. PMID- 11940143 TI - Clinical, genetic and microbiological findings in a Brazilian family with aggressive periodontitis. AB - BACKGROUND/AIM: Aggressive periodontitis comprises a group of rapidly progressive forms of periodontitis. Besides bacteria, a high level of subject susceptibility must be involved in the expression of disease. In the present study, we report the clinical, microbiological and genetic profile of a 14-individual family with aggressive periodontitis. METHOD: PCR was utilized to detect pathogenic bacteria of affected sites. DNA was obtained from epithelial cells through a mouthwash with 3% glucose and scrapping of the oral mucosa. RFLP-PCR was used to analyze cytokine genetic polymorphisms. RESULTS: Localized aggressive periodontitis was diagnosed for an 18-year-old systemically healthy non-smoking proband, with siblings displaying aggressive periodontitis. Bacteroides forsythus and Treponema denticola were the most frequent pathogens. The proband presented Actinobacillus actinomycetemcomitans and detectable levels of Porphyromonas gingivalis, Bacteroides forsythus and Treponema denticola. Allele 2 of IL-1alpha (-889) polymorphism was found in all individuals as well as allele 1 of the IL-1beta (+3953) gene. Alleles 1 and 2 (50 % each) of IL-1beta (-511), allele 1 of TNF alpha (-308) and allele 2 (in homo or heterozygosity) of IL-RN (intron 2) gene were present. CONCLUSION: The results show that the present microbiological and genetic parameters were not relevant for the prediction of periodontitis susceptibility in this family. PMID- 11940144 TI - Quantification of gingival edema using a new 3-D laser scanning method. AB - OBJECTIVES: The aim of the study was to investigate the suitability of measuring volume differences in the gingival tissue for monitoring changes in the inflammatory status of the gingiva. MATERIALS AND METHODS: Data for this investigation were obtained from a mouthrinse evaluation which was performed as a 4-week, double-blind, placebo-controlled, cross-over study in which localised experimental gingivitis was induced. 24 volunteers were enrolled in the study. Only the data from the placebo period of each subject were used in the current investigation. During the plaque accumulation periods, plaque guards were worn during routine performance of oral hygiene measures to prevent any plaque removal from the experimental area (1st and 2nd premolars and molars in one upper quadrant). Clinical examinations with assessment of plaque and gingivitis were performed on days 0, 4, 7, 14, 21, 28 and 42. Volume differences in the gingival papillae were determined between day 0 and days 21, 28, and 42, and between days 28 and 42 by taking measurements from replicas of the respective clinical situations using a 3-D laser scanner and reference-free automated 3-D superimposition software. Data were analysed with the Wilcoxon signed ranks test. RESULTS: Plaque accumulation in the experimental area resulted in a highly significant increase (p<0.001) of inflammation of the gingival papillae. The mean (standard deviation) papillary GI at baseline was 0.23 (0.34) as compared to 1.22 (0.27) and 1.2 (0.31) on days 21 and 28, respectively. The mean increase in volume of all papillae as compared to baseline was 25,478 micrometer3 after 21 days and 24,210 micrometer3 after 28 days. After resuming a normal oral hygiene regimen, mean volume of the papillae decreased between days 28 and 42 by 19,250 micrometer3. CONCLUSION: With this novel method, gingival papillary edema can be quantified in vivo from replicas of the clinical situation. PMID- 11940145 TI - The relationship of some negative events and psychological factors to periodontal disease in an adult Swedish population 50 to 80 years of age. AB - BACKGROUND: Clinical observations and epidemiological studies suggest that experiences of negative life events, especially those manifested as depression, may contribute to an increased susceptibility to periodontal disease. OBJECTIVE: In the present study, the prevalence of some negative life events and psychological factors and their relation to periodontal disease were investigated. The sample consisted of individuals 50-80 years of age from an extensive cross-sectional epidemiological study performed in 1993 in the city of Jonkoping, Sweden. METHOD: 298 dentate individuals from the Jonkoping study were randomly selected. Clinical and radiographic examinations included registration of the number of existing teeth, plaque index, gingival index, pocket depth, and alveolar bone loss. In addition, a questionnaire about socioeconomic status, life events, and psychological and stress-related factors was used. RESULTS: The results revealed that, in addition to the well-documented periodontal disease risk factors such as increased age, oral hygiene status, and smoking, the loss of a spouse (being a widow or widower) and the personality trait of exercising extreme external control were also associated with severe periodontal disease. CONCLUSION: The findings support recent studies suggesting that traumatic life events such as the loss of a spouse may increase the risk for periodontal disease. Above all, the present results indicate that an individual's ability to cope with stressful stimuli (coping behavior), as measured by the beliefs of locus of control of reinforcements may play a role in the progression of periodontal disease. PMID- 11940146 TI - Oral health and cardiovascular disease in Sweden. AB - OBJECTIVE: The purpose of the present study was to assess the relation between dental health and cardiovascular disease in an adult Swedish population. MATERIAL AND METHOD: A questionnaire was sent to 4811 randomly selected Swedes. It contained 52 questions about dental care habits, oral health, cardiovascular disease and their socio-economic situation. Odd ratios for all cardiovascular diseases (CVD) and the subgroup myocardial infarction, stroke and high blood pressure were calculated with a logistic regression model adjusted for age, gender, smoking, income, civil status and education. These ratios were calculated for subjects > or =41 years since few people suffer from CVD before this. RESULTS: The national questionnaire was answered by 2839 (59%) people between 20 84 years of age and, of them, 1577 were 41 years of age or more. We found a significant association between self-reported bleeding gums (odds ratio 1.60, p=0.0017), presence of dentures (odds ratio 1.57, p=0.0076) and known CVD, which has also been reported in international studies. However, no association between loose teeth, deep pockets and known CVD was detected. CONCLUSION: The results indicate that oral health and, especially gingival inflammation is associated with CVD. PMID- 11940147 TI - Subgingival microbial profiles in refractory periodontal disease. AB - BACKGROUND/AIM: The purpose of the present investigation was to examine subgingival microbial profiles associated with refractory periodontitis and to seek such profiles in periodontally healthy, periodontally well-maintained elder and untreated periodontitis subjects. METHODS: 36 subjects were defined as refractory on the basis of further attachment loss after scaling and root planing, surgery and systemically administered antibiotics. A total of 890 subgingival plaque samples (mean/subject=24.7) were taken from the mesial aspect of each tooth in each subject at baseline and individually processed for their content of 40 subgingival taxa using checkerboard DNA-DNA hybridization. Cluster analysis was performed on mean within subject species counts using the chord coefficient and an average unweighted linkage sort. Significant differences among clusters for individual and complexes of species were sought using the Kruskal Wallis test. The microbial profiles of the refractory subjects were compared with those of 27 periodontally healthy subjects (n plaque samples=708), 35 periodontally well-maintained elder subjects (n plaque samples=801) and 115 untreated adult periodontitis subjects (n plaque samples=2871). RESULTS: 28 of 36 refractory subjects fell into 4 clusters with >29% similarity. 10 of 40 species and 4 of 7 complexes differed significantly among clusters. Profile (Cluster) I (n=4) was characterized by high proportions of "yellow" and "green" complex species, profile II (n=3) by low total counts and high proportions of "orange" and "purple" complex species, profile III (n=9) by high total counts and counts of Actinomyces and "purple" complex species, profile IV (n=12) by high proportions of "red" and "orange" complex species. The mean profiles of each cluster were subjected to cluster analysis with microbial data from 4380 (mean 24.7) baseline subgingival plaque samples from 27 periodontally healthy, 35 treated, well-maintained elders and 115 untreated adult periodontitis subjects. 12 clusters were formed with >41% similarity. 3 of the refractory profiles were detected in 3 cluster groups. Profile II in a cluster of 1 healthy, 1 elder and 4 untreated periodontitis subjects; profile III in a cluster of 1 healthy, 2 elder and 12 periodontitis subjects; Profile IV, with 1 healthy and 5 untreated periodontitis subjects. The profile not detected in non refractory subjects was dominated by Streptococcus species. 9 clusters did not harbor refractory profiles. 11.1% of healthy, 8.6% of elder and 18.3% of periodontitis subjects were in clusters exhibiting refractory microbial profiles. CONCLUSIONS: 4 subgingival microbial profiles were detected among refractory subjects. "Refractory microbial profiles" could be detected in subjects who had not yet exhibited refractory disease. PMID- 11940148 TI - Metastatic breast carcinoma mimicking periodontal disease on radiographs. AB - BACKGROUND: Tumors can metastasize to the jaws, oral soft tissues, or salivary glands. Metastasis from distant organs and tissues to the jaws or oral cavity is considered rare. RESULTS: The mandible is the most common site of metastasis of the oral and maxillofacial region. Radiographically, there is usually an area of radiolucency with hazy outline. Such lesions are manifested as radiopaque or sclerotic areas. CONCLUSIONS: Radiographic view of a tumor in the present case mimicked a periodontal lesion with an osteolytic area around the roots. PMID- 11940150 TI - The M. tuberculosis antigen 85 complex and mycolyltransferase activity. AB - AIMS: The antigen 85 complex (Ag85) from Mycobacterium tuberculosis consists of three abundantly secreted proteins (FbpA, FbpB and FbpC2) which play a key role in the pathogenesis of tuberculosis and also exhibit cell wall mycolyltransferase activity. A related protein with similarity to the Ag85 complex was recently annotated in the M. tuberculosis genome as FbpC1. An investigation was carried out to determine whether FbpC1 may also possess mycolyltransferase activity, a characteristic feature of the Ag85 complex. METHODS AND RESULTS: Heterologous expression of FbpA, FbpC1 and FbpC2 was performed in Escherichia coli. Recombinant proteins were purified under non-denaturating conditions and used in an in vitro mycolyltransferase assay. CONCLUSIONS: In contrast to FbpA and FbpC2, recombinant FbpC1 did not possess in vitro mycolyltransferase activity and was not recognized by two monoclonal antibodies to the native Ag85. SIGNIFICANCE AND IMPACT OF THE STUDY: Mycolyltransferase activity is restricted to FbpA, FbpbB and FbpC2 only; the actual function of FbpC1 remains to be established. PMID- 11940151 TI - Ribotyping of Clostridium perfringens from industrially produced ground meat. AB - AIMS: Clostridium (Cl.) perfringens is a common cause of food poisoning outbreaks. Ribosomal DNA analysis (ribotyping), a method which analyses restriction fragment length polymorphisms in the chromosomal genes that encode rRNA, has been shown to be useful for microbial species identification and subtyping. METHODS AND RESULTS: The current study has used ribotyping to examine 111 Cl. perfringens isolates from industrially produced ground meat in order to collect a basis for a contamination survey. Among the 111 isolates 107 distinctly different ribopatterns were detected. In only four cases two Cl. perfringens isolates showed an identical ribopattern. The isolates gave identical ribotype patterns in three different runs, carried out 3-4 months apart from each other. CONCLUSIONS, SIGNIFICANCE AND IMPACT OF THE STUDY: The discriminatory index for EcoRI ribotyping of the Cl. perfringens isolates was 0 x 99. Results showed that ribotyping is suitable for subtyping Cl. perfringens isolates from raw meat. Ribotyping appeared to be a useful tool for profound epidemiologic studies of Cl. perfringens-contamination in food production and processing. PMID- 11940152 TI - Expression of the mel gene from Pseudomonas maltophilia in Bacillus thuringiensis. AB - AIMS: The objective of this work was to express a novel mel gene, responsible for melanin formation, in Bacillus thuringiensis. METHODS AND RESULTS: A novel mel gene from Pseudomonas maltophilia was sub-cloned into B. thuringiensis using a shuttle vector plasmid and electroporation. Results revealed that the mel gene was expressed under the control of the CryIIIA promoter in B. thuringiensis and conferred u.v. protection on the recipient strain. CONCLUSIONS: The novel mel gene from Ps. maltophilia expressed in B. thuringiensis conferred u.v. protection on the recipient strain. SIGNIFICANCE AND IMPACT OF THE STUDY: Products containing B. thuringiensis for pest control are sensitive to u.v. degradation. As melanin has the ability to act as a u.v. absorber, a recombinant B. thuringiensis strain producing melanin provides a new stability for B. thuringiensis preparations. PMID- 11940153 TI - The ability of the plasmid-encoded restriction and modification system LlaBIII to protect Lactococcus lactis against bacteriophages. AB - AIMS: To investigate the potential of the plasmid-encoded restriction and modification (R/M) system LlaBIII to protect Lactococcua lactis against bacteriophages during milk fermentations. METHODS AND RESULTS: The R/M system LlaBIII on plasmid pJW566 was cloned with a chloramphenicol cassette, resulting in plasmid pJK1. When introduced into L. lactis strains, pJK1 conferred increased phage resistance against the three most common lactococcal phage species 936, c2, and P335 and three unclassified industrial phages. The growth of the strains in RSM was not affected by the presence of plasmid pJK1. CONCLUSIONS: The plasmid encoded R/M system LlaBIII has great ability to protect L. lactis strains against bacteriophages in milk fermentations. SIGNIFICANCE AND IMPACT OF THE STUDY: This study evaluates the ability of the LlaBIII R/M system to function as a phage defence mechanism which is an essential step prior to considering utilizing it for improving starter cultures. PMID- 11940154 TI - Specific layers in aerobically grown microbial granules. AB - AIMS: To determine the optimal size of aerobically grown granules for wastewater treatment by measuring specific layers within the granules. METHODS AND RESULTS: A variety of biological layers were detected by oligonucleotide probes, specific fluorochromes, and fluorescent microspheres. The channels in the granule matrix penetrated to depths of 900 microm. A layer of obligate anaerobic bacteria was detected at a depth of 800 microm below the granule surface. Dead cells were also observed in the granule interior. CONCLUSIONS: Aerobically grown granules contained layers of aerobic and anaerobic micro-organisms. SIGNIFICANCE AND IMPACT OF THE STUDY: The optimal diameter of the aerobic granule is less than 1600 microm. This is twice the distance from the granule surface to the anaerobic layer. This approach can be used to optimize the thickness of other microbial aggregates such as flocs, colonies and biofilms. PMID- 11940155 TI - Arsenic (III) oxidizing Microbacterium lacticum and its use in the treatment of arsenic contaminated groundwater. AB - AIMS: To develop a microbially-assisted process for the removal of arsenic from contaminated groundwater. METHODS AND RESULTS: A culture of Microbacterium lacticum oxidizing up to 50 mmol l(-1) arsenic (III) was isolated from municipal sewage by an enrichment culture technique. Using culture immobilized on brick pieces and packed in a glass column, complete oxidation of As (III) from groundwater could be quickly achieved at neutral pH and ambient temperature with methanol as substrate. The oxidized As species were removed from groundwater using three different methods: zero valent iron, activated charcoal and ferric chloride. CONCLUSIONS: The oxidation of groundwater As (III) by a M. lacticum immobilized column, followed by its removal using activated carbon, could be an efficient method for the treatment of As (III)-contaminated groundwater. SIGNIFICANCE AND IMPACT OF THE STUDY: The study will be useful in developing a combined microbiological-chemical process for treating arsenic-contaminated groundwater. PMID- 11940156 TI - Development of a simple and sensitive fluorimetric method for isolation of coumaphos-hydrolysing bacteria. AB - AIMS: To develop a simple, rapid and sensitive fluorimetric assay to detect, isolate and characterize a soil bacterium capable of degrading the organophosphorus pesticide, coumaphos. METHODS AND RESULTS: A high throughput microtitre plate-based method was used to quantify coumaphos hydrolysis by the bacterium. The fluorescent hydrolysis product of coumaphos, chlorferon, was detected at levels as low as 10 nmol l(-1). Incorporation of coumaphos into agar plates allowed the rapid detection of coumaphos-hydrolysing bacteria when exposed to an excitation wavelength of approximately 340 nm. The coumaphos-hydrolysing enzyme could be visualized when bacterial cell extracts were separated on SDS PAGE, incubated with coumaphos and exposed to an excitation source as above. CONCLUSIONS: This method is 100-fold more sensitive than the currently used spectrophotometric method for coumaphos. SIGNIFICANCE AND IMPACT OF THE STUDY: This is a unique and versatile tool to screen for bacteria possessing phosphotriesterase activity. PMID- 11940157 TI - The correlation method for rapid monitoring of Escherichia coli in foods. AB - AIMS: A new rapid method was developed to rapidly monitor Escherichia coli counts in foods. MATERIALS AND RESULTS: One ml of modified selective broth with 4 methylumbelliferyl beta-D-glucuronide and 1 ml of food sample were mixed in a sterile test tube and incubated at 37 degrees C. The positive reaction (fluorescence under u.v. light) was monitored at regular 30 min intervals. The positive reaction times in test tubes were compared with actual E. coli numbers from tested samples. The growth of E. coli in test tubes (broth) was much faster than growth on agar. The first experiment was performed to evaluate the rapid correlation method using pure E. coli cultures. The correlation between E. coli counts by the conventional plating method and positive reaction (fluorescence production) times in test tubes was highly agreeable (r(2) = 0 x 95). In the case of low E. coli numbers, such as 2 x 0 log10 cfu ml(-1), the rapid correlation method detected their presence after 10 h incubation. When highly contaminated samples were assayed (8 log10 cfu ml(-1)), the rapid correlation method detected the presence of E. coli after 4 h incubation. In the ground beef experiment, the correlation between fluorescence production time and actual E. coli numbers was also strongly agreeable (r(2) = 0 x 92). CONCLUSIONS: From these results, it is obvious that the new rapid method can rapidly monitor E. coli counts in foods. SIGNIFICANCE AND IMPACT OF THE STUDY: The results indicated that the new method saved about 10-14 h incubation time compared to conventional plating methods. The rapid correlation method required much shorter incubation times compared to conventional plating methods for monitoring E. coli. PMID- 11940158 TI - Comparison of LST + MUG broth technique and conventional method for the enumeration of Escherichia coli in foods. AB - AIMS: To reduce the analysis time needed for the enumeration of Escherichia coli, a rapid fluorogenic method (MUG) which takes only 48 h was compared with the standard most probable number (MPN) method which takes 6 days as described in the International Standards Organization (ISO). This study provides reliability data for the fluorogenic method applied to certain foods. METHODS AND RESULTS: Both methods were applied to 500 food samples which were analysed for E. coli enumeration. Agreement between the two methods was found in 409 (81 x 8%) samples; 81 (16 x 2%) samples gave higher values by the fluorogenic method, and only 10 (2 x 0%) samples were more effectively assayed by the ISO method. According to statistical analysis, the reliability between the methods was r = 0 x 9706, r(2) = 0 x 9421 and Cronbach's alpha = 0 x 9851. While all three values showed a high degree of correlation (P < 0 x 0001) between the two methods, McNemar's test demonstrated a significant difference between them, indicating that the MUG method was more reliable than the ISO method. CONCLUSIONS: The data suggest that the fluorogenic method is more reliable and shorter to perform than the standard ISO method. SIGNIFICANCE AND IMPACT OF THE STUDY: Comparison of the two methods may provide a rapid and more reliable alternative for the enumeration of E. coli in food samples. PMID- 11940159 TI - Microfloral contamination and hydrolytic enzyme differences between monsooned and non-monsooned coffees. AB - AIMS: To analyse and compare, for the first time, fungal and bacterial populations and hydrolytic enzyme production in four monsooned and non-monsooned Arabica and Robusta coffee types. METHODS AND RESULTS: Overall, using serial dilution, the populations of bacteria (approximately 10(8) cfu g(-1)) were highest in monsooned coffees of both varieties. Fungal populations were lower (10(5) cfu g(-1)) and found predominantly on monsooned coffee beans. The major fungal species were Aspergillus terreus, A. restrictus and A. ochraceus. Of 19 semi-quantitative enzymes analysed, significantly higher concentrations of 2 naphthyl-butyrate, caprylate and 2-naphthyl-D-galactopyranosidase were present in Arabica monsooned coffee. CONCLUSIONS: The results suggest that speciality monsooned coffee has markedly different microbial and physiological characteristics from normally produced green coffees. SIGNIFICANCE AND IMPACT OF THE STUDY: Monsooned coffee may have a higher contamination with spoilage moulds, especially mycotoxigenic species. PMID- 11940160 TI - Does calculation of the 95th percentile of microbiological results offer any advantage over percentage exceedence in determining compliance with bathing water quality standards? AB - AIMIS: Draft WHO guidance is likely to suggest a calculated 95th percentile method, rather than percentage exceedence, in assessing compliance of bathing waters with microbiological standards. This study set out to determine whether this was an appropriate development. METHODS AND RESULTS: A series of Monte Carlo studies compared five non-parametric methods for calculating the 95th percentile with the parametric method and compares results with percentage exceedence. It is shown that the Hazen method gives the closest proximity to the parametric method for calculating 95th percentile values. However, the difference between 95th percentile results and percentage exceedence, as currently used, is trivial compared to uncertainty due to sample variation. CONCLUSIONS: It is concluded that a calculated 95th percentile for beach classification offers little advantage compared to percentage exceedence, other than offering a false sense of certainty. Furthermore, the additional calculation needed in determining 95th percentile values will demand electronic calculation, increase the chance of calculation errors and make the results less understandable to beach managers and the general public. SIGNIFICANCE AND IMPACT OF THE STUDY: This study should encourage the European Commission to keep percent compliance rather than calculation of 95th percentiles in any new bathing water directive. PMID- 11940161 TI - Cloning and sequencing of 16S rDNA and 16S-23S rDNA internal spacer region (ISR) from urease-positive thermophilic Campylobacter (UPTC). AB - AIMS: To clone and sequence the 16S rDNA and 16S-23S rDNA internal spacer region (ISR) from urease-positive thermophilic Campylobacter (UPTC). METHODS AND RESULTS: The primer sets for 16S rDNA and 16S-23S rDNA ISR amplified almost the full length of 16S rDNA and 16S-23S rDNA ISR. About 1500 bp for 16S rDNA and about 720 bp for 16S-23S rDNA ISR of the rrn operon of four strains of UPTC were identified after molecular cloning and sequencing. CONCLUSIONS: The four strains and CCUG18267 of UPTC showed approximately 99% sequence homology of 16S rDNA to each other, 96-97% to Camp. coli, 97-98% to Camp. jejuni and 97-98% to Camp. lari. SIGNIFICANCE AND IMPACT OF THE STUDY: For the first time, the nucleotide sequence of 16S-23S rDNA ISR of UPTC has been analysed. The sequence of ISR was almost identical among the four strains of UPTC. It is interesting that the UPTC intercistronic tRNAs demonstrated an order of tRNA of 5'-16S-tRNAAla-tRNAIle-23S 3' in the organisms. PMID- 11940162 TI - Improvement in the resting-cell bioconversion of penicillin G to deacetoxycephalosporin G by addition of catalase. AB - AIMS: To improve the resting cell bioconversion of penicillin G to deacetoxycephalosporin G (DAOG) by elimination of an oxidizing intermediate which inactivates the enzyme during the reaction. METHODS AND RESULTS: Resting cells of Streptomyces clavuligerus strain NP1 were incubated with penicillin G, required co-factors and decane in the presence of catalase or superoxide dismutase, and production of DAOG was measured. Catalase stimulated the bioconversion but superoxide dismutase did not. CONCLUSIONS: Production of hydrogen peroxide during the ring expansion reaction is at least partially responsible for enzyme inactivation. SIGNIFICANCE AND IMPACT OF THE STUDY: Catalase addition improves the bioconversion and will contribute to the eventual replacement of the current multi-step, expensive and environmentally-unfriendly chemical ring expansion by a biological route. PMID- 11940163 TI - Inhibition of biofilm formation and swarming of Bacillus subtilis by (5Z)-4-bromo 5-(bromomethylene)-3-butyl-2(5H)-furanone. AB - AIMS: (5Z)-4-Bromo-5-(bromomethylene)-3-butyl-2(5H)-furanone(furanone) of the marine alga Delisea pulchra was synthesized, and its inhibition of swarming motility and biofilm formation of Bacillus subtilis was investigated. METHODS AND RESULTS: Furanone was found to inhibit both the growth of B. subtilis and its swarming motility in a concentration-dependent way. In addition, as shown by confocal scanning laser microscopy, furanone inhibited the biofilm formation of B. subtilis. At 40 microg ml(-1), furanone decreased the biofilm thickness by 25%, decreased the number of water channels, and reduced the percentage of live cells by 63%. CONCLUSIONS, SIGNIFICANCE AND IMPACT OF THE STUDY: Natural furanone has potential for controlling the multicellular behaviour of Gram-positive bacteria. PMID- 11940164 TI - A liquid synchronized-growth culture assay for the identification of true positive and negative yeast three-hybrid transformants. AB - AIMS: To develop a simple and easy-to-use assay for detection of truely positive transformants from a yeast three-hybrid assay. METHODS AND RESULTS: The yeast three-hybrid system is a new powerful system for studying RNA-protein interactions in vivo. There are, however, many reports from investigators about the difficulty in distinguishing a positive from a negative result due to the hard-to-detect differences between truely positive transformants and the negative ones. A liquid synchronous-growth culture approach has been described for all positive and negative transformants and their growth densities have been compared to each other at fixed intervals of incubation times. We have designed a simple yet effective procedure to assay for positive and negative RNA-protein interactions based on liquid culture analysis of synchronously growing yeast cells. Results obtained from this new procedure clearly show differences in positive and negative transformants after a 24-h incubation of synchronously growing transformants in liquid culture. CONCLUSIONS: The procedure mentioned in this report shows clearly the differences between positive and negative results from a three-hybrid system. SIGNIFICANCE AND IMPACT OF THE STUDY: The method proposed here is a clear advantage over existing methods based on measuring growth on restrictive growth medium plates by the naked eye. This method will have substantial usage with investigators using the yeast three-hybrid studies for RNA-protein interactions. PMID- 11940167 TI - External examining--a challenge for new curricula? PMID- 11940165 TI - Rapid and specific identification of Shiga toxin-producing Escherichia coli in faeces by multiplex PCR. AB - AIMS: The object of this study was to develop a multiplex PCR system for rapid and specific identification of Shiga toxin-producing Escherichia coli (STEC) in faeces. METHODS AND RESULTS: A multiplex PCR (mPCR) protocol was developed using a primer pair specific for genes that are involved in the biosynthesis of the O157 E. coli antigen, and primers that identify the sequences of Shiga toxin 1 and 2 (stx 1 and stx1) and the intimin protein (eaeA). The mPCR assay was used for amplification of STEC genes in bacteria directly (after enrichment) in faeces. The test was very sensitive and could detect between 9 and 1 bacterial cells per gram of faeces. The mPCR was used for the examination of 69 bovine faecal samples derived from healthy cattle. The results indicated that 62 x 3% of the samples were positive, generating at least one PCR amplicon of the expected size. CONCLUSIONS: The method can be applied for rapid and specific identification of STEC bacteria in faecal samples, and for differentiation of their main virulence marker genes. SIGNIFICANCE AND IMPACT OF THE STUDY: The ability to sensitively detect Shiga toxin-producing E. coli directly in faeces within a short time represents a considerable advancement over more time consuming and less sensitive methods for identification and characterization of STEC bacteria. PMID- 11940168 TI - Assessment practices undermine self-confidence. PMID- 11940169 TI - Professional development in undergraduate medical curricula from an assessment point of view. PMID- 11940170 TI - Teamwork during education: the whole is not always greater than the sum of the parts. PMID- 11940171 TI - Chiropractors' attitudes to, and perceptions of, the impact of continuing professional education on clinical practice. AB - OBJECTIVES: As with other health care professions, there is a need for chiropractors to maintain their clinical competencies through continuing professional education (CPE) and development (CPD). This study set out to evaluate chiropractors' attitudes to, and opinions of, current CPE provision, and their perceived needs for the future. In particular, their perceptions of the impact of CPE on clinical practice were assessed. DESIGN: A survey in which primary quantitative and secondary qualitative data were collected using a self administered questionnaire. The questionnaire concentrated on four areas: (i) attitudes to CPE; (ii) impact of current CPE on practice; (iii) modes of delivery most likely to change practice; and (iv) future learning needs from continuing education programmes. RESPONDENTS: A random sample of chiropractors registered with the British Chiropractic Association. RESULTS: Overall, chiropractors were positive to CPE and cognisant of the need to keep up to date. However, in spite of several benefits, chiropractors did not perceive current CPE to be instrumental in changing practice. As might be expected, chiropractors perceived clinical updates and hands-on practical workshops to be most effective in changing clinical practice. DISCUSSION: There is currently considerable debate surrounding the ability of CPE and CPD to actually make a difference to the way clinicians practice. This study describes a method of evaluating the impact of CPE/D in changing clinical practice from the viewpoint of the practitioners themselves. PMID- 11940172 TI - Helping tomorrow's doctors to gain a population health perspective - good news for community stakeholders. AB - INTRODUCTION: The modernising agenda of the NHS and recommendations of professional bodies demand that all doctors achieve a basic understanding of a population health perspective. The principle of integrated learning and the logistics of provision make it inappropriate for such learning to be delivered solely by public health specialists, and community-based learning has been promoted as the best setting in which to assist this objective. However, there is little evidence from practice as to whether non-specialist staff are willing or able to play a role in orienting tomorrow's doctors to the needs of communities. METHODS: Semi-structured interviews, questionnaires and focus groups iterated the opinion of key stakeholders on their preferred contributions to community oriented undergraduate medical education. Framework analysis was used to elicit key outcomes and process factors. RESULTS: There was consensus that community based learning should be a core element of the medical curriculum as it can demonstrate the socioenvironmental context of care and the doctor's role in interagency working and preventive care. Effective academic/NHS partnerships were called for, with higher education leading the agenda on aims and objectives, creating collaborative structures, and reallocating resources to support new learning. Community-based stakeholders would offer the translation of theory into practice by demonstrating clinical and social diversity, models of teamworking, and a context for the application of prior learning. They also promoted recurrent contact with communities, and suggested an enhanced role in mentoring students through longer term relationships. CONCLUSION: Community-based personnel and NHS users are an under-used resource for medical education. Their proposed contributions strongly accord with the agenda for a population health perspective in basic training. Curriculum planners need to make long-term partnerships with community-based agencies, rather than using them as an intermittent provider of limited learning sessions with narrowly defined objectives. PMID- 11940173 TI - Learning in groups: Teamshaping in the teaching of medical psychology. AB - OBJECTIVES: Although it is of growing importance in modern medicine, the ability to work in teams usually plays no explicit role in medical education. We compared the method of Teamshaping with 'traditional' teaching. DESIGN: Ninety-six students in 6 groups pursued a course in medical psychology with traditional teaching methods, while 46 students in 4 groups participated in Teamshaping. To evaluate the team approach, we used the Group Climate Questionnaire, Short Version (GCQ-S). SETTING: University of Vienna, Medical School. SUBJECTS: Third year medical students. RESULTS: In terms of group climate both groups benefited from teaching, regardless of the method. Teamshaping, however, initially imposed a greater strain on the students than traditional teaching. This changed towards the end of the seminar. Teamshaping produced a stronger sense of commitment and a reduction of perceived conflicts. CONCLUSION: The model is recommended for teaching medical students because it not only extends traditional teaching towards small group learning but adds a competency in teamwork. It introduces students to a better working style and gives them an additional perspective for use in their future profession. PMID- 11940174 TI - 'It teaches you what to expect in future . . . ': interprofessional learning on a training ward for medical, nursing, occupational therapy and physiotherapy students. AB - AIM: This paper presents findings from a multimethod evaluation of an interprofessional training ward placement for medical, nursing, occupational therapy and physiotherapy students. CONTEXT: Unique in the UK, and following the pioneering work at Linkoping, the training ward allowed senior pre-qualification students, under the supervision of practitioners, to plan and deliver interprofessional care for a group of orthopaedic and rheumatology patients. This responsibility enabled students to develop profession-specific skills and competencies in dealing with patients. It also allowed them to enhance their teamworking skills in an interprofessional environment. Student teams were supported by facilitators who ensured medical care was optimal, led reflective sessions and facilitated students' problem solving. METHODS: Data were collected from all groups of participants involved in the ward: students, facilitators and patients. Methods included questionnaires, interviews and observations. RESULTS AND DISCUSSION: Findings are presented from each participating group, with a particular emphasis placed on the perspective of medicine. The study found that students valued highly the experiential learning they received on the ward and felt the ward prepared them more effectively for future practice. However, many encountered difficulties adopting an autonomous learning style during their placement. Despite enjoying their work on the ward, facilitators were concerned that the demands of their role could result in 'burn-out'. Patients enjoyed their ward experience and scored higher on a range of satisfaction indicators than a comparative group of patients. CONCLUSIONS: Participants were generally positive about the training ward. All considered that it was a worthwhile experience and felt the ward should recommence in the near future. PMID- 11940175 TI - Translating learning principles into practice: a new strategy for learning clinical skills. AB - AIM: There are data to suggest that medical school may not adequately prepare doctors for practice and that there are deficiencies in undergraduate teaching of skills in history taking, physical examination, diagnosis and management (clinical skills). There is a need to re-evaluate methods by which we can teach clinical skills effectively. This aim of this review was to describe the literature concerning the important principles underpinning effective clinical learning. Subsequently a structured learning tool and teaching process was developed in order to support these principles. METHOD: The principles of effective clinical learning were derived after a search of the medical education and relevant behavioural science literature. Consequently, a structured learning tool and teaching process was developed in order to potentiate the translation of these principles into practice for medical school training in clinical skills. RESULTS: Ten principles were derived from the 68 articles referred to in this review. These were: making active decisions, an individual focus to learning, gaining experience, feedback to the learner, reciprocal learning, holistic care, relevant learning, feasibility, cost efficiency and mentoring. A process for history taking, physical examination and management plan was developed for medical students which incorporated these principles. CONCLUSION: Relevant literature can provide the foundations for teaching and learning methods in medical education. We plan to trial this method and evaluate the impact on student learning outcomes. PMID- 11940176 TI - Professional development in undergraduate medical curricula--the key to the door of a new culture? AB - CONTEXT: One of the most pressing requirements for contemporary medical education is to develop a framework for theory and practice of professional development which results in the attainment of professional competencies suitably robust for a lifetime's practice. The proposed content of a professional development curriculum may be reasonably straightforward to establish from policy documents and public expectations, but the process of achieving the desired outcomes is more complex, because professional development is largely based on attitudinal learning. Attitudes are at the interface between the personal and public psyche, relying more on individual experience and the accumulated impact of social and cultural interpretations than on propositional knowledge, and are therefore less amenable to factual or didactic teaching. AIMS: The purpose of this discussion paper is to develop thinking on the conceptual frameworks which need to underpin curriculum decision-making for professional development, especially in undergraduate medical training where models of good practice are less well established. It brings together work from educational, sociological and psychological perspectives to elucidate the key principles which are most likely to result in acquisition of desirable professional attributes. IMPLICATIONS: The literature suggests that successful professional development needs to be based on explicit values, which are repeatedly demonstrated in the learning environment, and modelled by senior colleagues and tutors; that the curriculum should incorporate a clear model of emotional as well as cognitive development; should be a major component of summative assessment; and needs to include formative mentorship at all stages of training. PMID- 11940177 TI - Professional development for general practitioners through fellowship by assessment. AB - BACKGROUND AND STUDY AIMS: The Royal College of General Practitioners has designed its Fellowship by Assessment (FBA) programme with the twin goals of promoting higher standards of care and offering GPs a professional challenge. By November 2000, 223 eligible doctors (2%) had opted to take FBA. This number is increasing annually. There is, however, little research to account for why GPs undertake it, or what the barriers might be. The aim of our study was to investigate GPs' experience and thinking on this issue. METHODS: A total of 13 GPs who had attained Fellowship by Assessment in the Tamar and Severn Faculties were invited to participate in a qualitative study. They were identified from the list of RCGP Fellows in the Institute of General Practice, University of Exeter, and purposefully selected to include trainers and non-trainers, men and women, rural and urban practitioners, and single-handed and large practices. Analysis of in-depth interviews was informed by grounded theory. FINDINGS: FBA served to acknowledge high standards of care. There was some variation in views about FBA, however, ranging from a perspective which prized the professional award, to the view that endorsed it as a pragmatic and useful way of structuring and monitoring improvements. Fellowship by Assessment improves the care of patients and empowers the doctors by improving their confidence. It is, however, hugely time-consuming and while our findings suggest it should be encouraged, there is a need for ways to be found whereby doctors may be supported in this form of professional development. PMID- 11940179 TI - Context-bound communication skills training: development of a new method. AB - OBJECTIVES: To examine how communication skills training might be integrated into everyday clinical practice in a manner that is acceptable to clinicians. DESIGN: General practitioners from 3 group practices agreed to take part, in turn, in a study of how to manage difficult consultations about antibiotic prescribing for acute respiratory infections. This provided the opportunity to conduct communication skills training in which lessons learned from one practice were taken into the next. SETTING: United Kingdom general practices. SUBJECTS: Three groups of general practitioners. FINDINGS: Difficulties with the acceptability of a traditional off-site workshop approach, using role play as the main teaching method, led to the development of a new training method (context-bound training), which proved to be practical and acceptable to experienced clinicians. The main features of the method were the delivery of training in the clinicians' place of work, and the transformation of their reported difficult cases into scenarios which they then encountered with a standardized simulated patient before and after brief seminars. Everyday clinical experience was kept in the foreground and 'communication skills' in the background. CONCLUSIONS: The method is acceptable to clinicians and adaptable to a range of clinical situations. It offers potential for improving the communication skills of clinicians both in hospital and primary care settings. PMID- 11940178 TI - Assessing communication skills of GP registrars: a comparison of patient and GP examiner ratings. AB - CONTEXT: General practice. OBJECTIVES: To compare ratings of GP registrars' communication skills by patients and GP examiners. DESIGN: A comparative study where the communication skills of GP registrars were assessed both by patients, using a validated tool called the Doctors' Interpersonal Skills Questionnaire (DISQ), and by GP examiners as part of the Fellowship examination of the Royal Australian College of General Practitioners (RACGP). PARTICIPANTS: These included 138 GP registrars, 6075 patients, and more than 70 GP examiners. RESULTS: Spearman rank correlations were used to test the strength of the relationship between Fellowship examination and DISQ scores. Findings showed that there were several communication skills areas with mild (but significant) correlations between patient and GP examiner ratings. These areas included warmth of greeting, listening skills, respect, and concern for the patient as a person. No significant correlations were detected for explanation skills. Interestingly, the correlations between GP examiner and patient ratings were stronger for female GP registrars. CONCLUSION: There is some evidence that patients' ratings of GP registrars' communication skills is aligned to ratings made by GP examiners as part of the summative RACGP Fellowship examination. However, further work is required to assess the strength of this alignment, given that patient-doctor communication is assessed more widely through new components of the examination. PMID- 11940180 TI - Medical students' note-taking in a medical biochemistry course: an initial exploration. AB - Beginning medical students spend numerous hours every week attending basic science lectures and taking notes. Medical faculty often wonder whether they should give students pre-printed instructors' notes before lectures. Proponents of this strategy argue that provided notes enhance learning by facilitating the accurate transmission of information, while opponents counter that provided notes inhibit students' cognitive processing or even discourage students from attending lectures. Little if any research has directly addressed medical students' note taking or the value of providing instructors' notes. The educational literature does suggest that taking lecture notes enhances university students' learning. University students perform best on post-lecture testing if they review a combination of provided notes and their own personal notes, particularly if the provided notes follow a 'skeletal' format that encourages active note-taking. PMID- 11940182 TI - Standard setting in an objective structured clinical examination: use of global ratings of borderline performance to determine the passing score. PMID- 11940185 TI - Values of attending physicians: an evidence-based analysis of the current challenges. PMID- 11940187 TI - Using electronic mail as a method of surveying medical students' opinions and attitudes. PMID- 11940188 TI - Information sheets and consent forms in medical education research. PMID- 11940189 TI - A brief review of the pre-clinical curriculum of the BP Koirala Institute of Health Sciences, Dharan, Nepal. PMID- 11940190 TI - Ethics approval for research in medical education. PMID- 11940191 TI - Evidence-based admissions may limit ambitions to widen access. PMID- 11940197 TI - Molecular Pathology of the Prions. PMID- 11940192 TI - The hidden cost of Membership examinations. PMID- 11940199 TI - Professor P. O. Yates (1921-2001). PMID- 11940200 TI - The World Health Organization classification of malignant lymphoma: incidence and clinical prognosis in HTLV-1-endemic area of Fukuoka. AB - New insights into the pathogenesis of lymphoid malignancies have been gained through novel genetic, molecular and immunological techniques. A new classification system for lymphoid malignancies, known as the new World Health Organization (WHO) classification, has been proposed recently based on these findings. The relative incidence of the subtypes of malignant lymphoma is known to differ according to geographic location. Adult T-cell leukemia/lymphoma (ATLL) is a human malignancy associated with human T-cell leukemia virus type 1 (HTLV 1), and the Kyushu islands are an HTLV-1 endemic area. To clarify the relationship between the histological classification and prognosis of lymphoid malignancies, we reclassified previous cases in our department and summarized our previous reports using the WHO classification. Of 933 cases of lymphoid malignancies, 471 (50%) were B-cell lymphoma, 396 (42%) T/natural killer (NK) cell lymphoma and 41 (4%) Hodgkin lymphoma (HL). Analysis of clinical outcome showed favorable prognosis for HL, intermediate for B-cell lymphoma and poor prognosis for T-cell lymphoma. Among B-cell lymphomas, the commonest type was diffuse large B-cell lymphoma (n = 281; 60%). Marginal zone lymphoma of mucosa associated lymphoid tissue (MALT) was diagnosed in 82 cases (17%), follicular lymphoma in 52 (11%) and mantle cell lymphoma in 24 (5%). Other less common lymphomas were Burkitt lymphoma (n = 9; 2%) and lymphoblastic lymphoma (n = 5; 1%). Using overall survival rates, the various B-cell lymphoma types could be divided into three broad groups for prognostic purposes: (i) low-risk group comprising follicular lymphoma and MALT; (ii) intermediate-risk group comprising diffuse large B-cell lymphoma and Burkitt lymphoma; and (iii) high-risk group comprising mantle cell lymphoma and lymphoblastic lymphoma. Among the T/NK-cell lymphomas, the commonest type was ATLL (n = 191; 48%), followed by peripheral T cell lymphoma, unspecified (n = 83; 21%), angioimmunoblastic lymphadenopathy with dysproteinemia (AILD) (n = 38; 10%), anaplastic large cell lymphoma (ALCL) (n = 22; 6%). Less common types were lymphoblastic lymphoma (n = 17; 4%), nasal and nasal-type NK/T-cell lymphoma (n = 17; 4%), mycosis fungoides (MF) (n = 9; 2%) and other rare types. With respect to clinical prognosis, T/NK-cell lymphomas fell into three groups: (i) relative low-risk group comprising ALCL, AILD, MF and lymphoblastic lymphoma; (ii) relative intermediate-risk group comprising NK/T cell lymphoma and unspecified lymphoma; and (iii) extremely high-risk group comprising ATLL. Among the lymphoblastic lymphomas, B-cell type and T-cell type lymphomas exhibited different clinical outcomes. We conclude that the histological, phenotypic and genotypic classification of the new WHO system should be beneficial for the clinical approach to these tumors. PMID- 11940201 TI - E-cadherin expression in follicular carcinoma of the thyroid. AB - Follicular carcinoma (FC) of the thyroid is distinguished from follicular adenoma (FA) by confirmation of invasion or metastasis. However, it is still unknown how FC acquires the potential for invasion or metastasis. Twenty early FC (pT1 and pT2) were analyzed for immunohistochemical E-cadherin (E-CD) expression. Four of four (100%) widely invasive FC showed reduced E-CD expression, whereas only two of 16 minimally invasive FC showed reduced expression. In most of the minimally invasive FC, both E-CD and beta-catenin were expressed on cell-cell boundaries, even in areas where capsular invasion occurred. These results suggest that the mechanism of invasion may be different between widely invasive FC and minimally invasive FC, and that E-CD is probably responsible for the invasion of widely invasive FC. Analysis of methylation of the E-CD gene promoter using the methylation-specific polymerase chain reaction (MSP) method revealed no methylated alleles in the DNA from FC. PMID- 11940202 TI - Overexpression of osteopontin in hepatocellular carcinoma. AB - Human hepatocellular carcinoma (HCC) is one of the most common and aggressive malignancies. In order to identify genes involved in HCC progression, we conducted a differential display analysis and found osteopontin (OPN) to be overexpressed in HCC. OPN is known to be a secreted adhesive glycoprotein, associated with tumorigenesis and metastasis in several cancers. Quantitative polymerase chain reaction analysis of 30 HCC cases revealed the average ratio of OPN to glyceraldehyde-3-phosphate dehydrogenase in tumors to be significantly higher than that in the surrounding non-cancerous liver (4.7 +/- 1.6 vs 0.18 +/- 0.04, P = 0.0072). Immunohistochemistry confirmed the OPN protein was expressed mainly on cancer cells, and was positive in 12 of 30 HCC, most of which showed transcript overexpression. Both OPN transcript and OPN protein were significantly overexpressed in HCC with capsular infiltration, compared with HCC without capsular infiltration. Moreover, OPN-positive cancer cells were often dispersed in the periphery of cancer nodules and were adjacent to stromal cells. Although OPN overexpression was not related to vascular invasion or intrahepatic metastasis, OPN was suggested to play a role in HCC, especially in cancer-stromal interactions. PMID- 11940203 TI - CD34-positive stromal cells and alpha-smooth muscle actin-positive stromal cells in the tumor capsule of skin sweat gland neoplasms. AB - To elucidate the roles of CD34-positive stromal cells and alpha-smooth muscle actin-positive stromal cells at the tumor border of skin sweat gland neoplasms, we examined expression of stromal cell markers in the tumor capsule of 19 skin sweat gland neoplasms (16 mixed tumors of the skin and three nodular hidradenomas) using monoclonal antibodies to CD34, CD31, cytokeratin 14 (CK14), alpha-smooth muscle actin (ASMA) and high molecular weight caldesmon (HCD). We regarded CD34-positive, CD31-, CK14-, ASMA- and HCD-negative stromal cells to be CD34-positive stromal cells, and ASMA-positive, HCD-, CK14-, CD34- and CD31 negative stromal cells to be ASMA-positive stromal cells. CD34-positive stromal cells were detected in the tumor capsule of all 19 of the tumors examined. In nine of the 16 mixed tumors (56%) and all of the three nodular hidradenomas, ASMA positive stromal cells were detected at the immediate inner side of the CD34 positive stromal cell layers. These results indicate that cellular components in the tumor capsules of mixed tumors of the skin and nodular hidradenomas are CD34 positive stromal cells and ASMA-positive stromal cells, and suggest that stromal cells of these two cell types are associated with tumor capsule formation of skin sweat gland neoplasms. PMID- 11940204 TI - Herpesvirus (HSV-1, EBV and CMV) infections in atherosclerotic compared with non atherosclerotic aortic tissue. AB - The viral nucleic acid of herpes simplex virus type 1 (HSV-1), Epstein-Barr virus (EBV) and cytomegalovirus (CMV) was studied by polymerase chain reaction (PCR), Southern blotting and in situ hybridization (ISH) in aortic tissues from 33 autopsies. In 23 cases involving persons who ranged from 23 weeks to 75 years of age at the time of death, the tissue was histologically non-atherosclerotic. Of these 23, aortic tissues tested positive for HSV-1 in 13%, for EBV in 13% and for CMV in 4%. In the other 10 cases involving persons who were 53-75 years old at death, atherosclerotic aortic tissue tested positive for HSV-1 in 80%, for EBV in 80% and for CMV in 40%. Neither double nor triple infections occurred in the non atherosclerotic group, whereas six of 10 were positive for two viruses, and two of 10 were positive for three viruses in the atherosclerotic group. By in situ hybridization, the viruses were localized in cells morphologically consistent with endothelial cells and smooth muscle cells. We detected HSV-1, EBV and CMV DNA in cells in the upper portion of the non-atherosclerotic aortic wall, whereas viral DNA was detected more extensively in atherosclerotic lesions than in non atherosclerotic tissue. We also are the first to show the existence of EBV DNA in the human aortic wall. In conclusion, we suggest that the high incidence and kinds of herpesviruses are related to the high incidence of atherosclerosis. PMID- 11940205 TI - Acceleration of murine AA amyloidosis by oral administration of amyloid fibrils extracted from different species. AB - We herein report that experimental murine amyloid A (AA) deposition is accelerated by oral administration of semipurified amyloid fibrils extracted from different species. Three groups of mice were treated with semipurified murine AA amyloid fibrils, semipurified bovine AA amyloid fibrils or semipurified human light chain-derived (A(lambda)) amyloid fibrils for 10 days. After 3 weeks, each mouse was subjected to inflammatory stimulation by subcutaneous injection with a mixture of complete Freund's adjuvant supplemented with Mycobacterium butyricum. The mice were killed on the third day after the inflammatory stimulation, and the spleen, liver, kidney and gastrointestinal tract were examined for amyloid deposits. Amyloid deposits were detected in 14 out of 15 mice treated with murine AA amyloid fibrils, 12 out of 15 mice treated with bovine AA amyloid fibrils and 11 out of 15 mice treated with human A(lambda) amyloid fibrils. No amyloid deposits were detected in control mice receiving the inflammatory stimulant alone or in amyloid fibril-treated mice without inflammatory stimulation. Our results suggest that AA amyloid deposition is accelerated by oral administration of semipurified amyloid fibrils when there is a concurrent inflammatory stimulation. PMID- 11940206 TI - Alpha-fetoprotein-producing lung carcinoma: report of three cases. AB - Three cases of alpha-fetoprotein (AFP)-producing lung carcinoma were studied histologically and immunohistochemically. Samples were obtained from two men and one woman who ranged in age from 64 to 71 years. Serum AFP levels for the three samples were 9826, 74.4 and 24.3 ng/mL. One case was classified as stage IIIA and two as stage IIIB. Two cases were diagnosed as large cell neuroendocrine carcinoma, and AFP expression was detected immunohistochemically. One of these samples showed differentiation to a hepatoid carcinoma, while the other was combined with a squamous cell carcinoma. The remaining case was a squamous cell carcinoma, and AFP was detected in only some of the tumor cells. All patients died within 2 years. The Ki-67 labeling indices of the AFP-producing pulmonary carcinomas (30.2 +/- 4.6%) were significantly higher than those of AFP-negative pulmonary carcinomas (P < 0.05). The high proliferative activity, advanced stage at presentation, vascular endothelial growth factor expression and vascular invasion observed in these tumors may explain the poor prognosis of AFP-producing lung carcinomas. PMID- 11940207 TI - Papillary carcinoma of the thyroid gland forming a myofibroblastic nodular tumor: report of two cases and review of the literature. AB - We encountered two unique cases of papillary carcinoma of the thyroid gland forming fibroma-like nodular tumors. The two cases showed well-defined nodules composed largely of fibromatosis-like stroma with small foci of papillary carcinoma. This type of tumor is relatively rare and has been reported with the name of 'papillary carcinoma of the thyroid with fibromatosis-like stroma or nodular fasciitis-like stroma'. The stromal cells have had myofibroblastic features in all reported cases, including the present cases. The pathogenesis of this tumor is still unknown. We must be careful to differentiate this type of thyroid tumor from fibrosarcomatous anaplastic carcinoma, particularly in the intraoperative consultation. We speculate that this unique tumor is an autonomous proliferation of stromal cells, based on the immunohistochemical analyses of early stromal changes in usual-type papillary carcinoma of the thyroid. The present names of this disease may not represent the proper features of this tumor because all the reported cases formed fibroma-like nodules. We propose the name 'papillary carcinoma forming myofibroblastic nodular tumors'. PMID- 11940208 TI - Chordoma of cervical vertebra protruding into the oral cavity. AB - A 70-year-old man was admitted to hospital because of oral and cervical masses. Computed tomographic scanning revealed a lobulated mass lesion in the retropharyngeal region, with a protruding extension in the oral cavity and with destruction of the second cervical vertebra. A biopsy was performed under the diagnosis of a retropharyngeal tumor. Histologically, this lesion was composed of vacuolated tumor cells in a solid or cord-like arrangement, with an abundant myxoid matrix. Immunohistochemically, the tumor cells were positive for pancytokeratin, epithelial membrane antigen and S-100 protein. The tumor was diagnosed as chordoma. Chordoma presenting as an intra-oral mass lesion is very rare. PMID- 11940209 TI - Expression of parathyroid hormone-related protein (PTHrP) in multiple myeloma. AB - Multiple myeloma is a plasma cell neoplasia often associated with multiple skeletal lesions and hypercalcemia. Several cytokines, including interleukin (IL) 1, IL-6 and tumor necrosis factor-beta (TNF-beta), derived from myeloma cells are thought to accelerate osteoclastic bone resorption and cause hypercalcemia through a paracrine mechanism. We report on a case of a 69-year-old man with multiple myeloma associated with hypercalcemia and advanced osteolytic lesions. After bisphosphonate treatment and MP (melphalan and prednisolone) therapy, the patient's serum calcium level was successfully but transiently recovered to the normal range. Biochemical analysis showed a remarkable increase in serum parathyroid hormone-related protein (PTHrP; 3.7 pmol/L) and IL-6 (22.0 pg/mL). On the other hand, parathyroid hormone and 1alpha,25(OH)2 vitamin D3 were suppressed. By immunohistochemistry and in situ hybridization on aspiration biopsied bone marrow clot sections, PTHrP mRNA and protein were detected in the cytoplasm of myeloma cells. The rate of PTHrP-positive myeloma cells was estimated to be at least one-third. Since PTHrP can, as an endocrine factor, systemically act on bone and kidney, hypercalcemia in this case might have been caused through both local osteolytic hypercalcemia and humoral hypercalcemia of malignancy mechanisms. PMID- 11940210 TI - Perivascular myoma: case report with immunohistochemical and ultrastructural studies. AB - A subcutaneous myopericytoma-type perivascular myoma arising in the elbow of a 61 year-old woman is described. The tumor was well demarcated and consisted microscopically of small ovoid and spindle cells arranged in a concentric fashion, surrounding small to medium-sized vessels and imparting a superficial resemblance to hemangiopericytoma. In some areas, the cellular whorls were separated by myxoid stroma. Cells located between cellular whorls appeared immature with scant cytoplasm but did not show distinct nuclear anaplasia, increased mitoses or foci of necrosis. Immunohistochemistry showed that constituent cells were positive for alpha-smooth muscle actin and desmin. Electron microscopy disclosed that the immature-looking cells, as well as the ovoid and spindle-shaped cells, possessed focal densities along with thin filaments, subplasmalemmal densities, pinocytotic vesicles and an external lamina. These ultrastructural and immunohistochemical features indicate a myoid nature of pericytic cells and justify this type of neoplasm being categorized as perivascular myoid tumor. PMID- 11940211 TI - Epithelioid trophoblastic tumor of the uterus: cytological and immunohistochemical observation of a case. AB - Epithelioid trophoblastic tumor (ETT) is a new entity of trophoblastic tumor and 14 such cases were reported by Shih and Kurman in 1998. However, only three subsequent cases supporting ETT have been reported. Recently, we experienced a case of ETT in a 37-year-old woman whose preoperative endometrial brushings showed atypical mononucleate giant cells and who underwent hysterectomy with the diagnosis of a uterine fibroid. The specimens revealed a 2.5 x 3.0 cm yellow-tan intramural nodule located in the lower uterine segment, which was composed of a neoplastic proliferation of intermediate trophoblasts in epithelioid arrangements. Immunohistochemically, the tumor cells were diffusely positive for cytokeratin and inhibin-alpha, and focally positive for human chorionic gonadotropin and human placental lactogen. She presented an uneventful clinical course as of September 2001. PMID- 11940212 TI - CD97, CD95 and Fas-L clearly discriminate between chronic pancreatitis and pancreatic ductal adenocarcinoma in perioperative evaluation of cryocut sections. AB - It is a major problem to distinguish between pancreatitis and pancreatic adenocarcinoma when it comes to the perioperative evaluation of pancreatic cryocut sections. In this respect, pathologists are showing a steadily growing interest in the potential application of apoptotic and dedifferentiation factors as diagnostic and prognostic markers. This study investigated the mRNA and protein expression of CD97, CD95 and Fas-L in snap-frozen material obtained from human pancreatic ductal adenocarcinomas (PDC; n = 50), tissues from pancreatitis (PT; n = 40) and normal pancreatic tissues (PN; n = 36). Reverse transcription polymerase chain reaction (RT-PCR) analysis revealed that CD97, CD95 and Fas-L mRNA were expressed on a similarly high level in all tissues. In contrast, short time immunohistochemical evaluation showed that the CD95 protein was strongly expressed in PT and PN, but not in PDC. Fas-L protein was expressed strongly in PDC, whereas only weak or no expression was noted in PT or PN. CD97 protein expression was detected only in PT and in poorly differentiated PDC. Our data demonstrate that CD97, CD95 and Fas-L can be used as additional markers to distinguish between pancreatitis and pancreatic duct cell carcinoma in cryocut sections. PMID- 11940213 TI - Emerging multifunctional aspects of cellular serine proteinase inhibitors in tumor progression and tissue regeneration. PMID- 11940214 TI - Aberrant expression of pRb, p16, p14ARF, MDM2, p21 and p53 in stage I adenocarcinomas of the lung. AB - Cancers are always associated with cell cycle abnormalities. To clarify the cell cycle abnormalities present in lung adenocarcinomas, various cell cycle regulatory proteins of both the pRb and p53 pathways were studied immunohistochemically in 50 cases of stage I adenocarcinoma of the lung. In regard to the pRb pathway, most adenocarcinomas showed frequent expression of both p16 and pRb proteins, and aberrant expression in the pRb pathway was observed in about one-quarter of stage I adenocarcinomas. In regard to the p53 pathway, the frequency of immunohistochemical positivity was 8% for p14ARF, 64% for MDM2, 20% for p53 and 24% for p21. In this pathway, the immunohistochemical profile of p14ARF-negative/MDM2-positive/p53-negative is characteristic of stage I adenocarcinoma of the lung. An inverse relationship was found between MDM2 and p53 protein and was associated with the differentiation status of stage I adenocarcinoma of the lung. Our results suggest that the disruption of the pRb and p53 pathways is frequently observed in the early stages of lung adenocarcinoma and might play an important role in the growth and differentiation of adenocarcinoma of the lung. PMID- 11940215 TI - Analysis of cell damage in Helicobacter pylori-associated gastritis. AB - Helicobacter pylori infection is currently considered to be a major cause of acute and chronic gastritis, and of gastric and duodenal ulcers. Superoxide dismutase (SOD) is well known for scavenging superoxide radicals such as reactive oxygen species (ROS), subsequently protecting cells from oxidative injury, and for maintaining tissue homeostasis. In this study, we therefore evaluated the level of SOD activity and protein expression, as well as various factors associated with oxidative injury, in H. pylori-positive (n = 46) and -negative (n = 28) gastric mucosa obtained from endoscopy, in order to elucidate the possible biological significance of SOD in these mucosa. Overall SOD activity was significantly higher in H. pylori-positive mucosa (15.5 +/- 7.0 U/mg protein) than in negative mucosa (9.2 +/- 10.6 U/mg protein), and decreased markedly following H. pylori eradication (8.2 +/- 4.2 U/mg protein). Enzyme-linked immunosorbent assay (ELISA) analysis of SOD revealed that the manganese SOD (Mn SOD) level in H. pylori-positive mucosa (1166.7 +/- 435.2 ng/mg protein) was significantly higher than in control tissues (446.3 +/- 435.3 ng/mg protein) and in mucosa obtained following eradication therapy (431.9 +/- 189.9 ng/mg protein). The level of Mn-SOD protein showed a significant correlation with degree of inflammation in the gastric mucosa. Moreover, Mn-SOD immunolocalization patterns were well correlated with the activity and protein levels evaluated by ELISA. Factors presumably associated with oxidative injury in human gastric mucosa, including terminal deoxynucleotidyl transferase-mediated dUTP-biotin nick-end labeling, Ki-67, 8-hydroxydeoxyguanosine and single-stranded DNA, were all significantly higher in H. pylori-positive gastric mucosa than in control tissue and in tissue following eradication. These results all suggest that Mn-SOD, but not cytoplasmic copper-zinc SOD, plays an important role as an anti-oxidant against ROS generated in H. pylori-infected gastric mucosa and, subsequently, in the maintenance of cell turnover in gastric mucosa. PMID- 11940216 TI - Histogenesis and morphogenesis of depressed-type early colorectal carcinoma. AB - The aim of this study was to elucidate the histogenesis and morphogenesis of depressed-type early colorectal carcinomas (DECas). Eighty-seven DECas were selected for examination. The 87 DECas included 23 (26.4%) absolutely depressed (ABS) types (depressed mucosa being thinner than the normal mucosa; histologically true depression), 40 (46.0%) relatively depressed (REL) types (the thickness of the depressed mucosa being the same or greater than that of normal mucosa, with depression being relative compared to marginal elevation) and 24 (27.6%) ulcerated (UL) types (no remnant intramucosal component in the depressed area). The rates of residual adenomatous components were 0/23 (0%) for ABS type and 12/40 (30%) for REL type, indicating that the ABS type was thought to have arisen de novo and the REL type was thought to have arisen de novo and via an adenoma-carcinoma sequence. In 13 of the 23 (56.5%) ABS types, carcinoma was located within the depressed area alone, suggesting that many ABS types show a depression from their initial stage. While 33 of the 40 (82.5%) REL types accompanied by marginal elevation consisted of neoplastic mucosa, in 19 of the 33 (57.6%) cases, the depressed area showed a higher grade of atypia than the area of marginal elevation. This suggested that the depression in the REL type was generated secondarily at the center of a pre-existing, superficially elevated tumor due to progression of its histological grade of atypia. ABS type was speculated to be the precursor of UL type since the two types showed similarity (no statistical significant difference) in percent depression (69.6 vs 64.3%), grade of atypia of carcinoma (52.2 and 79.2% consisted of carcinoma with high grade atypia alone), and frequency of being accompanied by non-neoplastic reactive marginal elevation (56.5 vs 70.8%). PMID- 11940217 TI - Evaluation of interobserver agreement in scoring immunohistochemical results of HER-2/neu (c-erbB-2) expression detected by HercepTest, Nichirei polyclonal antibody, CB11 and TAB250 in breast carcinoma. AB - To examine interobserver reproducibility in the judgments of HER-2/neu (c-erbB-2) oncoprotein expression, HER-2/neu expression was scored immunohistochemically. A total of 106 breast carcinomas were independently scored by five or six pathologists, using four commercially available anti-bodies (HercepTest, Nichirei polyclonal antibody, CB11 and TAB250). Two systems were used for the evaluation. In the Dako system, the intensity of membrane staining was primarily evaluated whereas in the Tohoku system, the area of the membrane-staining-positive cells was primarily evaluated. All four antibodies gave excellent concordance rates among the pathologists' judgments of the scores. Using the HercepTest and Nichirei polyclonal antibody, dividing the cases into two groups designated score 3+ and score 0/1+/2+ achieved a higher percentage of concordant cases than division into groups designated score 2+/3+ and score 0/1+. Using the HercepTest, the Dako system tended to achieve higher concordance than the Tohoku system. Conversely, CB11 and TAB250 showed similar concordance rates under both the Dako and the Tohoku systems, regardless of how the cases were divided. High interobserver agreement levels were achieved only when HER-2/neu overexpression was defined as a score of 3+, using any of the antibodies; these were similar between the Dako system and the Tohoku system. Positivity by TAB250 was lower than that by CB11. Using the HercepTest and Nichirei polyclonal antibody, cases with a score of 3+ almost always achieved complete interobserver agreement and showed HER-2/neu gene amplification. Conversely, cases with a score of 2+ achieved a low percentage of interobserver agreement and showed no gene amplification. Because judgment of a case as score 2+ was not very reproducible, and a score of 3+ was of biological significance and had high interobserver reproducibility, we suggest that only a score of 3+ be considered as overexpression. When cases of score 2+ are entered into a protocol, evaluation should be given by multiple observers and/or by using other tests such as fluorescence in situ hybridization. PMID- 11940219 TI - Hepatoid adenocarcinoma in Barrett's esophagus associated with achalasia: first case report. AB - We report an unusual hepatoid adenocarcinoma in Barrett's esophagus with achalasia, which developed in a 44-year-old Japanese woman. The patient received an esophago-gastrectomy after diagnosis of the tumor and achalasia at the lower esophagus, 4 months before her death due to multiple metastatic tumors of the liver. The main granular tumor removed surgically was a hepatoid adenocarcinoma, mainly composed of clear cancer cells (alpha-1 antitrypsin, albumin and alpha fetoprotein positive), with elements of choriocarcinoma and tubular adenocarcinoma. Non-neoplastic specialized columnar epithelium was present extensively near the oral side of the tumor edge in the esophagus, indicating Barrett's esophagus. This unusual tumor was therefore considered to have originated in Barrett's esophagus. The gastroesophageal reflux was presumed to have occurred for a long period, as there was a well-preserved fundic gland in the stomach and a history of frequent vomiting from the patient's youth, accounting for the appearance of achalasia. PMID- 11940218 TI - Overexpression of MDM2 in a sclerosing epithelioid fibrosarcoma: genetic, immunohistochemical and ultrastructural study of a case. AB - Sclerosing epithelioid fibrosarcoma (SEF) is an extremely rare soft-tissue neoplasm. Here, we describe the molecular genetic alterations and histological, immunohistochemical and ultrastructural features of a primary SEF arising in the retroperitoneum. The tumor consisted of uniform small round to ovoid epithelioid cells, arranged in nests and cords and surrounded by a prominent hyalinized collagenous matrix. The tumor cells expressed only vimentin. Ultrastructurally, the tumor cells showed features of fibroblasts, with an abundant rough endoplasmic reticulum in the cytoplasm. Neither p53 gene mutations nor p53 protein overexpression were detected, but more than 70% of all tumor cells showed strong immunoreactivity with murine double minute 2 (MDM2). Our results suggest that MDM2 overexpression is likely to play a role in tumorigenesis in this lesion in p53-dependent or p53-independent pathways. To our knowledge, the present study is the first molecular genetic study of this rare lesion. Further studies will be necessary to clarify the molecular basis of tumorigenesis of this rare lesion. PMID- 11940220 TI - Diffuse pagetoid squamous cell carcinoma of the esophagus combined with choriocarcinoma and mucoepidermoid carcinoma: an autopsy case report. AB - Esophageal squamous cell carcinoma in situ (SCCIS) with diffuse pagetoid features is a recently recognized rare variant of squamous cell carcinoma. A histopathological study of a specimen from a 70-year-old male Japanese patient is reported. The patient died of respiratory failure due to rapidly progressing metastatic pulmonary tumors of unknown origin 73 days after the onset of hemosputum. Autopsy disclosed widespread metastasis of choriocarcinoma in the absence of tumors of the testes or other common sites of germ cell tumors. Elevation of human chorionic gonadotropin (hCG-beta) levels was later detected in the stored serum. Serial histological evaluation of the entire esophagus revealed a small primary site of choriocarcinoma in a background of diffuse SCCIS, mainly of pagetoid type, accompanied by several small foci of submucosally invasive squamous cell carcinoma and primary mucoepidermoid carcinoma. These stimulated nodal metastasis independently of the choriocarcinoma. The SCCIS did not alter the gross mucosal appearance. This is the first reported case of diffuse pagetoid SCCIS combined with choriocarcinoma. Morphological findings and previous studies suggest that the extensive SCCIS of the esophagus resulted from pagetoid spread of tumor cells. The invasive squamous cell carcinoma, mucoepidermoid carcinoma and choriocarcinoma are suggested to have originated from the overlying SCCIS. PMID- 11940221 TI - Mixed neuronal-glial tumor of the digestive tract: distinctive entity from gastrointestinal stromal tumor? AB - A 53-year-old-woman presenting with pelvic discomfort was found to have a 9.5 cm tumor located in the wall of the ileon. Light microscopy showed that the tumor was made of fascicles of plump spindle cells and bizarre epithelioid cells. A cuff of lymphoid cells was also present at the tumor margin. The tumor cells strongly expressed tau protein, neuron-specific enolase, protein green product 9.5 and glial fibrillary acid protein (GFAP), but did not show positive immunostaining for S-100 protein, CD34 or CD117. The tumor showed unequivocal ultrastructural evidence of neural differentiation. Skeinoid fibers were scattered throughout the tumor. This is the first mixed neuronal-glial tumor of the digestive tract to be described in the literature. Such histological and immunohistochemical features could be misinterpreted as features of digestive schwannoma. We suggest that this tumor is distinct from gastrointestinal stromal tumors in lacking CD34 and CD117 expression. PMID- 11940222 TI - Primary osteosarcoma of the uterine corpus: case report and review of the literature. AB - A rare case of rapidly growing osteosarcoma that developed in the uterine corpus of a 62-year-old woman is presented. The tumor occupied almost the entire pelvic cavity and extended into the abdominal cavity, with marked involvement of the intestines. Histopathologically, the tumor was composed of an osteoblastic component, accompanied by conspicuous bone formation, and a fibroblastic component. The tumor cells were positive for vimentin and osteocalcin, as well as desmin, alpha-smooth muscle actin and muscle-specific actin, but negative for h caldesmon. The results indicated myofibroblastic differentiation in a part of the tumor. A review of 14 reported cases and our case of uterine osteosarcoma revealed that this tumor has a biologically aggressive nature, although its histopathological and immunohistochemical features are similar to those of osteosarcomas in soft tissue and bone. As the prognosis of patients with this tumor is poor, it is of importance to differentiate this tumor from other types of tumors arising from the uterine corpus. PMID- 11940223 TI - Uterine tumors resembling ovarian sex-cord tumors producing parathyroid hormone related protein of the uterine cervix. AB - A case of uterine tumors resembling ovarian sex-cord tumors (UTROSCT) producing parathyroid hormone-related protein (PTH-rP) of the uterine cervix is reported. A 66-year-old woman underwent total hysterectomy with bilateral salpingo oophorectomy due to the possibility of a malignant uterine tumor. A fairly well circumscribed tumor, measuring 8 x 5 x 7 cm, was present in the myometrium of the cervix and extended into the endocervical mucosa. Histologically, the tumor showed predominantly sex-cord-like differentiation and the features of conventional endometrial low-grade stromal sarcoma were observed in part. Immunohistochemically, the tumor cells were negative for CD10. From these findings, we diagnosed the present case as Clement and Scully's group II UTROSCT arising from the uterine cervix. To our knowledge, this is the first report of the cervical occurrence of UTROSCT. Furthermore, in this tumor, production of PTH rP was demonstrated by normalization of serum PTH-rP after the tumorectomy and immunoreactivity for PTH-rP in the tumor cells. PMID- 11940224 TI - Apocrine adenocarcinoma of the eyelid with aggressive biological behavior: report of a case. AB - A case of apocrine adenocarcinoma of the eyelid that showed unusually aggressive biological behavior is reported. The patient was a 57-year-old man who complained of discomfort and excessive lacrimation of the left eye. A subcutaneous tumor measuring 2.5 cm was found at the medial canthus of the upper eyelid, and a plica like subconjunctival spread was noted in the lacrimal caruncle. Invasion into the extraocular muscles and metastasis to the cervical lymph nodes and bone were already present at the time of initial presentation. Histopathologically, the tumor showed features of poorly differentiated adenocarcinoma, and polygonal tumor cells had large, hyperchromatic nuclei with prominent nucleoli and abundant eosinophilic cytoplasm. The formation of ductal structures was found occasionally. The differentiation of the tumor cells towards the apocrine gland was corroborated by immunohistochemistry using monoclonal antibodies GCDFP-15 and B72.3. The histogenesis and pathological differential diagnosis are discussed briefly, and the tumor was considered to have originated in the Moll's gland in the eyelid. This case emphasizes that apocrine adenocarcinomas of the ocular region have the potential for aggressive biological behavior, including distant metastasis. PMID- 11940226 TI - Novartis Prizes for Immunology 2001, Stockholm. PMID- 11940227 TI - NK cells, MHC class I molecules and the missing self. AB - This article is based on a lecture presented at the Novartis Prize ceremony at the International Congress of Immunology in July 2001. It gives a personal and historical perspective on the research performed by the author and his colleagues during the development and pursuit of the model of 'missing-self recognition' for natural killer (NK) cells. This model is based on the idea that one important function of NK cells is to detect and eliminate cells because they fail to express normal self markers. Further mechanistic models predicted the existence of inhibitory major histocompatibility complex (MHC) class I specific receptors, later identified by the fellow Novartis laureates contributing in this issue. The article covers the first decade (1980-1990) of research on this concept. It discusses factors contributing to the formulation of a hypothesis, the use of predictions and experimental test models, the importance of international collaborations and reagent exchange, and several other aspects that allowed the progression of this research project. Finally, the perspective of today's knowledge is used to discuss some surprising findings where the missing-self hypothesis made the wrong predictions, or at least failed to make the correct ones. PMID- 11940228 TI - Natural killer cells: a mystery no more. AB - Recent years have witnessed remarkable progress in our understanding of the molecular mechanism regulating natural killer (NK) cell function. NK cells can sense whether cells have lost the surface expression of major histocompatibility complex (MHC)-class I molecules. The discovery of MHC-class I-specific inhibitory receptors clarified the basis of this discrimination and elucidated the nature of the 'off' signal. However, the receptors responsible for the 'on' signal in the process of natural cytotoxicity remained mysterious until recently. Here, we describe the identification and characterization of such receptors and discuss the emerging implications of these findings in different diseases. PMID- 11940229 TI - The search for the missing 'missing-self' receptor on natural killer cells. AB - In this article, I outline the studies that led to the discovery of Ly49A as the first 'missing-self ' receptor on natural killer cells. It is a more detailed and formal version of my acceptance speech at the Novartis Award ceremony at the International Congress of Immunology in Stockholm. Yet, it is impossible to mention my gratitude to all of my previous mentors and colleagues or to credit all the scientists and immunologists who preceded us by making contributions, large and small, to knowledge upon which our findings were based. It is therefore humbling to accept the 2001 Novartis Award for Basic Immunology along with Professors Klas Karre and Lorenzo Moretta. PMID- 11940230 TI - Primary immunodeficiency diseases: natural mutant models for the study of the immune system. AB - Primary immunodeficiency diseases represent many natural mutant models for the analysis of many aspects of immunology. Examples dealing with T- and B-cell receptors are herein discussed. They also represent privileged models in medicine, because well characterized monogenic inherited diseases can in some instances be viewed as good targets for gene therapy as shown here for severe combined immunodeficiency (SCID). PMID- 11940231 TI - Recognition of IgG-derived peptides by a human IgM with an unusual combining site. AB - A monoclonal immunoglobulin (Ig)M cryoglobulin (Mez) with interesting binding behaviour was isolated from a Waldenstrom's macroglobulinemia (WM) patient. It demonstrated very strong binding to peptides derived from the sequences of human IgG. However, when tested for binding to intact IgG, this antibody (Ab) did not show any rheumatoid factor (RF) activity. We propose several nonexclusive structural interpretations of the Mez-binding propensities, based on the orientations and solvent accessibilities of ligand residues and the nature of the Ab-binding site. To further characterize the structural features of Mez-peptide binding, IgG-derived octapeptides were docked into the Mez fragment variable (Fv) binding site, revealing additional reasons for Mez-binding selectivity based on the interactions of the docked peptides with the Mez Fv. The problem was also approached from an immunological perspective. Comparisons of Mez variable region of the light chain (VL)/variable region of the heavy chain (VH) sequences with those of human germlines and known IgM RFs allowed us to provide a possible outline tracing the structural and functional origins of the Mez IgM. Coupled with examinations of interactions in docked complexes, this analysis led us to propose that the potential for RF activity, demonstrated through Mez binding to IgG-derived peptides, was owing to the inherent sequence and structure of the Mez IgM, rather than to somatic mutations. Thus, Mez IgM may occupy an intermediate niche between IgMs with and without RF activity. PMID- 11940232 TI - The humoral response in TCR alpha-/- mice. Can gammadelta-T cells support the humoral immune response? AB - An optimal humoral response requires T-cell help; however, it has been questioned if this help comes exclusively from alphabeta-T cells or whether gammadelta-T cells also contribute. We have attempted to answer this question by studying the humoral response in T-cell receptor alpha-chain knockout (alpha-/-) mice, which lack the alphabetaT cell subset. Two model antigens were used to characterize the response: the thymus-independent (TI) antigen native dextran B512 (Dx), and the thymus-dependent (TD) antigen heat shock protein (HSP65) from Mycobacterium tuberculosis. When challenged with Dx, the alpha-/- mice elicited a strong antibody response and formed rudimentary germinal centres (GCs), a T-cell dependent reaction. In contrast, the humoral response to HSP65 was poor. However, alpha-/- mice became primed when challenged with HSP65, because when supplemented with wild-type thymocytes, the antigen-primed animals were able to mount a stronger response than the nonprimed ones when challenged with HSP65. A crucial step seems to be the collaboration between gammadeltaT cells and antigen presenting cells (APCs), as splenocytes from alpha-/- mice were able to respond to HSP65 in an environment containing primed-APCs. Based on these results, we propose a model for B-cell activation in the alpha-/- mice. PMID- 11940234 TI - Association of antigen specificity and migratory capacity of memory T cells in rheumatoid arthritis. AB - Among the T cell pool of multiple specificities in the rheumatoid synovial tissues (ST) we have previously shown a lack of proliferative response of T cells to Acanthamoeba polyphaga [1]. In contrast, peripheral blood (PB) derived T cells proliferate to the antigen. The aim of the present study was to establish whether there is a preferential migration of some T cell specificities to the joint in rheumatoid arthritis (RA) patients dependent on the chemokine system, and to identify which chemokine receptors are involved in the migratory process. For this purpose, PB-derived T cell lines and clones from RA patients specific for A. polyphaga, herpes simplex virus (HSV) and Campylobacter jejuni were developed. Their migratory capacities towards ST-derived chemokine supernatants were analysed. Expression of CCR1, CCR2, CCR5, CCR6, CCR7, CXCR3 and CXCR4 were analysed by FACS, and attracting chemokines were identified by blocking studies. We found that the migratory capacities of T cells specific for C. jejuni and HSV were markedly higher against synovial chemokines than T cells specific for A. polyphaga. CCR5 and CXCR3 were expressed by all high-migrating T cell lines and clones. CCR2 was expressed at higher levels on the high-migrating T cell lines compared with the low-migrating A. polyphaga lines. Neutralization of RANTES (regulated upon activation normal T cell expressed and secreted) in the ST cell derived supernatant reduced T cell migration of all T cell lines and clones by 60 90%, while neutralization of MCP-1 reduced the migratory capacity of CCR2 expressing T cells by 45-80%. In conclusion, the ability of T cells to migrate towards chemokines produced by ST cells is associated with the T cell specificity. Blocking of single chemokines substantially reduced the migratory capacity of memory T cells to ST cell-derived supernatant indicating unique roles for each chemokine receptor in the process of T cell migration. PMID- 11940233 TI - Differential expression of neurotrophic factors and inflammatory cytokines by myelin basic protein-specific and other recruited T cells infiltrating the central nervous system during experimental autoimmune encephalomyelitis. AB - Recent evidence suggests that autoimmune reactions in the central nervous system (CNS) not only have detrimental consequences but can also be neuroprotective, and that this effect is mediated by the expression of neuronal growth factors by infiltrating leucocytes. Here we dissect these two phenomena in guinea pig myelin basic protein peptide (gpMBP 63-88)-induced experimental autoimmune encephalomyelitis (EAE) in the Lewis rat. Real-time TaqMan polymerase chain reaction (PCR) was used to measure mRNA for the nerve growth factors, brain derived neurotrophic factor (BDNF) and neurotrophin (NT)-3. As reference, the well-known proinflammatory mediator molecules interferon (IFN)-gamma and tumour necrosis factor (TNF)-alpha were quantified. In whole lumbar cord tissue, both the nerve growth factors and the proinflammatory cytokines, IFN-gamma and TNF alpha, displayed similar expression patterns, peaking at the height of the disease. Among the infiltrating inflammatory cells isolated and sorted from the CNS, alphabeta+/T-cell receptor (TCR)BV8S2+, but not alphabeta+/TCRBV8S2-, recognized the encephalitogenic MBP peptide. Interestingly, these two populations displayed contrasting expression patterns of nerve growth factors and proinflammatory cytokines with higher inflammatory cytokine mRNA levels in alphabeta+/TCRBV8S2+ cells at all time intervals, whereas the levels of BDNF and NT3 were higher in alphabeta+/TCRBV8S2- cells. We conclude that a potentially important neuroprotective facet of CNS inflammation dominantly prevails within other non-MBP peptide-specific lymphoid cells and that there are independent regulatory mechanisms for neurotrophin and inflammatory cytokine expression during EAE. PMID- 11940235 TI - A partially humanized monoclonal antibody to human IFN-gamma inhibits cytokine effects both in vitro and in vivo. AB - The mouse monoclonal antibody (MoAb) IGMB17 (muIGMB17) is a high-affinity antibody- neutralizing human interferon (IFN)-gamma and, accordingly, is a potential therapeutic agent for patients suffering from various diseases in which the cytokine is abnormally expressed. The clinical usefulness of mouse antibodies is limited, however, owing to their immunogenicity in humans. MuIGMB17 antibody was partially humanized by engrafting a small portion of mouse light chain (LC) in a human framework and by engineering its heavy chain (HC) in a chimeric version. The engineered IGMB17 (huIGMB17) was able to replicate a range of functional properties of the original muIGMB17, namely, specific binding to IFN gamma, inhibition of histocompatibility complex (HLA-DR) expression in response to IFN-gamma induction, reversion of IFN-gamma antiproliferative activity on sensitive cell lines. We have hypothesized that as huIGMB17 was able to block IFN gamma binding to its receptor as well as its murine counterpart, huIGMB17 could neutralize all cytokine activity, also in vivo. Indeed huIGMB17 was capable of interfering with delayed-type hypersensitivity reaction in humans, thus demonstrating its effectiveness in neutralizing IFN-gamma-mediated reactions in vivo. PMID- 11940236 TI - Development of mixed Th1/Th2 type immune response and protection against Mycobacterium tuberculosis after rectal or subcutaneous immunization of newborn and adult mice with Mycobacterium bovis BCG. AB - Cell-mediated immunity plays a key role in containing the growth of Mycobacterium tuberculosis in the host. The induction of an antibody response or a mixed cell mediated and humoral response is frequently associated with tuberculosis disease or a decrease in the ability to control M. tuberculosis load. We recently reported the induction of similar immune responses and protection by rectal, subcutaneous (SC) or intradermal administration of Mycobacterium bovis BCG in adult mice, guinea pigs and macaques. The rectal immunization, which did not induce the side-effects associated with parenteral routes (axillary adenitis) and which could be used to reduce the risks of viral transmission associated with unsafe injections in the developing world, was analysed and compared in newborn and adult BALB/c mice. The rectal and SC immunization induced, in mice immunized as newborns or as adults, a mixed T helper 1/T helper 2 (Th1/Th2) immune response; however, particularly in adult mice, after SC administration of BCG, the level of Th2 immune response is significantly higher than it is by the rectal route. Six months after immunization with BCG, rectal and SC delivery induced similar levels of protective immunity against a virulent challenge with M. tuberculosis strain (H37Rv) in mice immunized as adults, but the rectal BCG delivery triggered stronger protection than the SC delivery if mice were immunized as newborns. PMID- 11940237 TI - A novel lacrimal gland autoantigen in the NOD mouse model of Sjogren's syndrome. AB - Recent research has demonstrated a crucial role for autoantibodies in the pathogenesis of Sjogren's syndrome (SS)-like disease in the non-obese diabetic (NOD) mouse, but it remains to be determined which antibody species among all those present are directly related to the various aspects of pathology. To identify autoantigens in the NOD mouse system, we have taken the approach of using immunoglobulin (Ig)G purified from sera of NOD mice exhibiting SS-like symptoms to screen cDNA expression libraries derived from exocrine gland mRNA. Here we report the identification of a novel autoantigen, designated LGP10, expressed in lacrimal and submandibular glands. Autoantibodies to this protein are prominent in the sera of NOD mice starting at 11-12 weeks of age, but not in control nonautoimmune mice. LGP10 has no known function, but bears similarities to various other proteins produced by epithelia. Interestingly, all of these similar proteins have been linked to immunosuppression and/or steroid binding, both processes that could have a significant impact on pathological features of SS. PMID- 11940238 TI - Diminished Th1 and Th2 cytokine production in healthy adult nonresponders to recombinant hepatitis B vaccine. AB - Vaccination of healthy adults with recombinant hepatitis B (rHB) vaccine fails to induce a protective antibody response in a proportion of individuals. Imbalanced T-helper (Th)1/Th2 response has been attributed to the lack of specific antibody response to rHB vaccine. In this study, in vitro production of interleukin (IL) 2, interferon (IFN)-gamma and IL-10 was investigated in Iranian healthy adults vaccinated with rHB vaccine. Peripheral blood mononuclear cells (PBMC) were isolated from 18 high responders and eight nonresponders and stimulated with rHB antigen or phytohaemaglutinin (PHA) mitogen. The cytokines were quantitated in culture supernatants by sandwich enzyme-linked immunosorbent assay (ELISA). Our results demonstrated a significant decrease in the production of IL-2, IFN-gamma and IL-10 (P < 0.005) in response to rHB antigen. The levels of all cytokines induced by PHA were similarly represented in both groups of vaccinees. These findings suggest that unresponsiveness to rHB vaccine may be owing to inadequate Th1- and Th2-like cytokine production. PMID- 11940241 TI - Uveitis and pupillary block glaucoma in an aphakic dog. AB - Unilateral uveitis with pupillary occlusion and secondary glaucoma was treated with neodymium:YAG laser iridotomy and iridencleisis in an aphakic 2-year-old male Miniature Schnuauzer. The dog presented 4 months after bilateral phacoemulsification with a complaint of blepharospasm of the left eye. Examination revealed anterior uveitis with pupillary occlusion and iris bombe. A slit-lamp mounted Q-switched Nd:YAG laser was used to create a central iridotomy to deepen the anterior chamber and an iridencleisis was performed to recreate a pupil. The eye remained visual and normotensive over a 6-month follow-up period. PMID- 11940242 TI - Advances in topical glaucoma therapy. AB - Significant advances have recently been achieved in the development of topical glaucoma medications. The primary advantage of a topical preparation is the reduced incidence of adverse systemic effects attributable to a given drug compared to its systemically administered counterpart. However, the strong protective barrier of the eye forces topical ophthalmic preparations to be highly concentrated and in some cases, they have the potential to produce unwanted systemic effects, particularly in smaller animals. Oral carbonic anhydrase inhibitors are commonly associated with adverse effects in both humans and animals. Two recently developed topical carbonic anhydrase inhibitors, dorzolamide and brinzolamide, have shown promise in reducing intraocular pressure in animals and systemic side effects are apparently limited with their use. The topical alpha2-agonist apraclonidine, on the other hand, effectively reduces intraocular pressure in cats and dogs, but in its currently available form is likely to induce unwanted systemic effects. Latanoprost is a topical prostaglandin F2alpha analog that has proven effective in reducing intraocular pressure in dogs and horses, but while systemic side effects have not yet been reported, this topical preparation may exacerbate pre-existing or concurrent ocular inflammatory disease. PMID- 11940243 TI - SLO angiography: arterio-venous filling times in monkey and minipig. AB - Confocal scanning laser ophthalmoscope (cSLO) is a new technique which enables ocular fundus image recording and dynamic retinal angiography to be performed. The ocular fundus image is acquired sequentially, point by point, and is reconstructed on a video monitor at the rate of 25 images per second. The aim of this paper is to evaluate the feasibility of measuring retinal arterio-venous filling times (AVFT) with a I + Tech cSLO. Three young adult cynomolgus monkeys and three young adult Gottingen minipigs were used as experimental models. All animals were anesthetized using a zolazepam + tiletamine mixture injected intramuscularly; heart rate and rectal temperature were monitored and corneal irrigation was regularly performed. For all subjects, prior to examination, hematocrit and globe axial length were measured. The images were recorded, stabilized and analyzed. The retinal examination consisted of retinal images with 40 degrees field cSLO, retinal fluorescein angiography and arterio-venous 50% filling time measurements. For each subject all images were easily recorded while keeping the animals in a normally lighted room without having to use any additional optical device. AVFT using an I + Tech cSLO is easily performed in monkeys and minipigs. AVFT measurements in minipigs and monkeys are similar. These results suggest that minipigs can replace monkeys as an experimental species for AVFT investigations. PMID- 11940244 TI - Partial orbital rim resection, mesh skin expansion, and second intention healing combined with enucleation or exenteration for extensive periocular tumors in horses. AB - Ocular and periorbital sarcoids and squamous cell carcinoma are common in equine practice. Extensive involvement of periorbital tissues often necessitates removal of the globe if the function of the eyelids can not be maintained with tumor removal alone. This report describes a modification of the standard enucleation or exenteration technique for cases in which there is insufficient skin to achieve primary closure following complete surgical excision. The caudal portion of the dorsal orbital rim is protuberant; partial excision with an osteotome facilitates skin closure by decreasing the size of the wound. Mesh expansion of skin via multiple rows or parallel stab incisions can also be used as an adjunct to facilitate closure. Four horses underwent enucleation or exenteration using the orbital rim resection and mesh skin expansion techniques for extensive periocular tumors that were unresponsive to prior treatments. Follow-up intervals ranged from 6 to 42 months and no horses had tumor regrowth. PMID- 11940245 TI - Treatment of granulomatous meningoencephalomyelitis in a dog. AB - The clinical work-up, diagnosis and follow-up of an 8-year-old, female-spayed Shih Tzu with diffuse, granulomatous meningoencephalomyelitis (GME)-causing visual deficits is reported. The use of cytosine arabinoside as an alternative treatment for GME is discussed. PMID- 11940246 TI - Amiodarone-induced keratopathy in healthy dogs. AB - Amiodarone has a broad spectrum as an antiarrhythmic agent and is indicated for patients with atrial and ventricular arrhythmias. Amiodarone-induced corneal deposits are the most common reversible side effect (70-100%) in humans. Additional ocular effects in humans include deposits in the lens, retina and optic nerve. This study was conducted to determine ocular effects of chronic oral amiodarone in healthy dogs. Six chronically amiodarone-treated dogs and four controls were used for this study. Ophthalmic examination was performed using biomicroscopy and indirect ophthalmoscopy at the end of 4th, 7th and 11th weeks when dogs received amiodarone. Corneal microdeposits were observed at the end of the 7th week in one eye and at end of the 11th week in the other eye of one dog. Immediately following euthanasia, corneas and optic nerves were harvested for light and electron microscopic analysis. Light microscopic analysis showed corneal deposits in the basal epithelial cells of the cornea of the clinically affected dog. In addition, a significant increase in basal cell turnover as indicated by mitotic index was observed in the affected dog compared to both nondeposit amiodarone and control groups. All remaining animals were normal. One out of six dogs treated with amiodarone demonstrated corneal deposits (16%). This prevalence is low compared to humans. Explanations for this may include species variations particularly in volume of lacrimal secretion, or the need for longer administration. In addition, sunlight is believed to exacerbate the corneal deposits in humans and all dogs in this study were housed indoors. PMID- 11940247 TI - Normal structure and age-related changes of the equine retina. AB - Investigations of the pathophysiology of ocular diseases require a detailed knowledge of the microanatomy of the eye. The available information is still inadequate for the equine retina despite the importance of eye diseases in equine medicine. Here we provide a comprehensive analysis of the histologic features of the horse eye as a reference for future studies. Thirty normal eyes of 15 healthy horses were examined immediately after slaughter. The retina of the horse differs considerably in the degree and quantity of neurons and glial elements as well as in vascular patterns compared to the retina of other domestic animals. Morphometric analysis revealed that the thickness of the retina varies between 80 microm at the ora serrata and 250 microm medial to the optic disc. Approximately 90% of the equine retina is comparatively thin (< 130 microm). This is a physiologic response to the distance that oxygen can diffuse in avascular retina. Ganglion cells form a single layer in all parts of the retina. The majority of ganglion cells are very large Nissl-positive cells. Small Nissl-negative ganglion cells are less abundant. A high ganglion cell density is found only in the central area. Vascularization is virtually absent from the retina with the exception of a narrow strip around the disc of the optic nerve, as revealed by lectin histochemistry. Light microscopy of the eyes of older horses repeatedly revealed cystoid degenerations in the retina adjacent to the pars plana of the ciliary body, as well as a destruction of the regular layering of the peripheral region of the retina. PMID- 11940248 TI - Pectinate ligament dysplasia and narrowing of the iridocorneal angle associated with glaucoma in the English Springer Spaniel. AB - OBJECTIVE: The aim of the study was to evaluate if tonometry and gonioscopy could serve as predictors of later glaucoma development in the English Springer Spaniel, a breed prone to developing primary glaucoma. ANIMALS STUDIED: Gonioscopy was performed on 279 English Springer Spaniel dogs, 119 males and 160 females, with clinically normal eyes. In addition, 14 dogs, five males and nine females were examined for glaucoma. RESULTS: A positive relation between pectinate ligament dysplasia (PLD) and narrowing of the relative width of the ciliary cleft (RWOCC) and also between PLD, RWOCC and age was demonstrated. The prevalence of PLD was 25.5%, which is higher than in other breeds reported. A positive association was demonstrated between PLD and glaucoma, as well as between narrowing of the RWOCC and glaucoma. Glaucoma was not observed in dogs with normal appearance of the iridocorneal angle. English Springer Spaniels related to dogs with glaucoma show more narrowing of the RWOCC and more PLD than unrelated dogs. CONCLUSION: This study demonstrates the positive association between PLD and glaucoma, between narrowing of the iridocorneal angle and glaucoma and the effect of age on the iridocorneal angle. Mating of dogs with normal iridocorneal angles appears to reduce the presence and degree of abnormal appearance of the iridocorneal angle in the offspring. However, breeding only dogs with normal iridocorneal angles without consideration of their relationship to dogs with glaucoma is not a guarantee for preventing glaucoma in the offspring. PMID- 11940250 TI - Congenital cataracts and persistent hyaloid vasculature in a llama (Lama glama). AB - A 9-month-old llama was evaluated for apparent blindness. Bilateral cataracts were diagnosed and cataract surgery was performed on the right eye. At the time of surgery persistent hyperplastic primary vitreous, persistent hyperplastic tunica vasculosa lentis, and a persistent hyaloid artery were observed. Prior to surgery Power Doppler ultrasound revealed a patent tunica vasculosa lentis OS. Despite reports of a poor success rate for llama cataract surgery, through use of careful tissue handling, phacoemulsification, viscoelastic endothelial protectants, anti-inflammatories, and BSS-Plus irrigating solution, vision was successfully restored in both eyes of the llama. PMID- 11940249 TI - Cyanoacrylate tissue adhesive for treatment of refractory corneal ulceration. AB - Isobutyl cyanoacrylate tissue adhesive (BCTA) was used in the treatment of refractory superficial corneal ulcers in 17 dogs, one cat, and one rabbit, present 2 weeks to 7 months (mean 6.8 weeks +/- 6.1) prior to referral. Little to no sedation was required in the majority of cases, with only topical anesthetic applied prior to debridement and BCTA application. The presence of the tissue adhesive caused mild discomfort for several days after application, as reported by the owners. The ulcers healed, and the tissue adhesive sloughed in approximately 3 weeks (+/- 1 week). Mild neovascularization of the cornea resolved with topical corticosteroids. The use of BCTA offers a simple, safe and noninvasive treatment for refractory corneal ulcers. PMID- 11940251 TI - Combined corneal lipid and calcium degeneration in a dog with hyperadrenocorticism: a case report. AB - Corneal degeneration may occur with a deposition of lipids or calcium, or both. Calcareous and lipid degeneration may be either primary or secondary, associated with systemic diseases such as primary hyperlipidemia, hyperlipidemia associated with hyperadrenocorticism, and hypothyroidism. The authors report a case of bilateral corneal lipid and calcium degeneration in a 7-year-old female Poodle with hyperadrenocorticism. The condition worsened with Lysodren therapy but responded to surgical excision. PMID- 11940252 TI - Different effects of rat interferon alpha, beta and gamma on rat hepatic stellate cell proliferation and activation. AB - BACKGROUND: Liver fibrosis is the common sequel of chronic liver diseases. Recent studies have identified hepatic stellate cells as the primary cell type mediating hepatic fibrogenesis. It has been demonstrated that hepatic stellate cells undergo a process of activation during the development of liver fibrosis. During the activation process, hepatic stellate cells acquire myofibroblast-like phenotype featuring the expression of smooth muscle alpha actin. Interferons have been employed for the treatment of viral hepatitis. However, it is unclear what is the effect of interferons on the prevention and treatment of liver fibrosis. Moreover, it is not clear whether there are any differences among interferon alpha, interferon beta, and interferon gamma in the treatment of liver fibrosis. Therefore, our objective in current study is to investigate the effects of rat interferon-alpha, interferon-beta, and interferon-gamma on the proliferation and activation of rat hepatic stellate cells. RESULTS: Rat interferon-beta and interferon-gamma significantly inhibited rat hepatic stellate cell proliferation while rat interferon-alpha did not affect the cell proliferation under the same culture condition. Inhibition of cell proliferation was confirmed by both WST-1 cell proliferation assay and 5-bromo-2'-deoxy-uridine incorporation assay. Similar results were observed regarding interferons regulation of hepatic stellate cell activation. Both rat interferon-beta and interferon-gamma reduced smooth muscle alpha-actin abundance after 6 days treatment, but rat interferon alpha did not alter smooth muscle alpha-actin level. CONCLUSIONS: Our results indicate that rat interferon-alpha and interferon-beta have different biological effects on rat hepatic stellate cells and suggest that there are different signaling events between interferon-alpha and interferon-beta in hepatic stellate cells. PMID- 11940253 TI - Activation of the peroxisome proliferator-activated receptor alpha protects against myocardial ischaemic injury and improves endothelial vasodilatation. AB - BACKGROUND: The peroxisome proliferator-activated receptor alpha (PPARalpha) plays an important role in the metabolism of lipoproteins and fatty acids, and seems to protect against the development of atherosclerosis. To evaluate the possible protective role of PPARalpha on cardiovascular function, the effect of the PPARalpha agonist, fenofibrate was assessed with respect to ischaemia/reperfusion injury and endothelial function in mice. RESULTS: Fenofibrate treatment reduces myocardial infarction size and improves post ischaemic contractile dysfunction. Hearts from PPARalpha null mice exhibit increased susceptibility to ischaemic damages and were refractory to protection by fenofibrate treatment suggesting that the beneficial effects of fenofibrate were mediated via PPARalpha. Furthermore, fenofibrate improves endothelium- and nitric oxide-mediated vasodilatation in aorta and mesenteric vascular bed. A decreased inhibitory effect of reactive oxygen species in the vessel wall accounts for enhanced endothelial vasodilatation. However, the latter cannot be explained by an increase in nitric oxide synthase expression nor by an increase sensitivity of the arteries to nitric oxide. CONCLUSIONS: Altogether the present data suggest that fenofibrate exerts cardioprotective effect against ischaemia and improves nitric oxide-mediated response probably by enhancing antioxidant capacity of the vessel wall. These data underscore new therapeutic perspectives for PPARalpha agonists in ischaemic myocardial injury and in cardiovascular diseases associated with endothelial dysfunction. PMID- 11940254 TI - Ethics and critical care in the new millennium. PMID- 11940256 TI - From bench to bedside: bacterial growth and cytokines. PMID- 11940257 TI - Early resuscitation in the emergency room: dramatic effects that we should not ignore. PMID- 11940258 TI - Rotating antibiotics in the intensive care unit: feasible, apparently beneficial, but questions remain. PMID- 11940259 TI - Translating research evidence into clinical practice: new challenges for critical care. PMID- 11940261 TI - Pro/con clinical debate: is high-volume hemofiltration beneficial in the treatment of septic shock? PMID- 11940260 TI - Pro/con clinical debate: the use of prone positioning in the management of patients with acute respiratory distress syndrome. PMID- 11940262 TI - ICUs worldwide: an overview of critical care medicine in South Africa. PMID- 11940263 TI - Clinical review: a paradigm shift: the bidirectional effect of inflammation on bacterial growth. Clinical implications for patients with acute respiratory distress syndrome. AB - Clinical studies have shown positive associations among sustained and intense inflammatory responses and the incidence of bacterial infections. We hypothesized that cytokines secreted by the host during acute respiratory distress syndrome may indeed favor the growth of bacteria and explain the association between exaggerated and protracted systemic inflammation and the frequent development of nosocomial infections. To test this hypothesis, we conducted in vitro studies evaluating the extracellular and intracellular growth response of three clinically relevant bacteria in response to graded concentrations of pro inflammatory cytokines tumor necrosis factor-alpha, IL-1beta, and IL-6. In these studies, we identified a U-shaped response of bacterial growth to pro inflammatory cytokines. When the bacteria were exposed in vitro to a lower concentration of cytokines, extracellular and intracellular bacterial growth was not promoted and human monocytic cells were efficient in killing the ingested bacteria. Conversely, when bacteria were exposed to higher concentrations of pro inflammatory cytokines, intracellular and extracellular bacterial growth was enhanced in a dose-dependent manner. The bidirectional effects of proinflammatory cytokines on bacterial growth may help to explain the frequent occurrence of nosocomial infections in patients with unresolving acute respiratory distress syndrome. PMID- 11940264 TI - Clinical review: severe asthma. AB - Severe asthma, although difficult to define, includes all cases of difficult/therapy-resistant disease of all age groups and bears the largest part of morbidity and mortality from asthma. Acute, severe asthma, status asthmaticus, is the more or less rapid but severe asthmatic exacerbation that may not respond to the usual medical treatment. The narrowing of airways causes ventilation perfusion imbalance, lung hyperinflation, and increased work of breathing that may lead to ventilatory muscle fatigue and life-threatening respiratory failure. Treatment for acute, severe asthma includes the administration of oxygen, beta2 agonists (by continuous or repetitive nebulisation), and systemic corticosteroids. Subcutaneous administration of epinephrine or terbutaline should be considered in patients not responding adequately to continuous nebulisation, in those unable to cooperate, and in intubated patients not responding to inhaled therapy. The exact time to intubate a patient in status asthmaticus is based mainly on clinical judgment, but intubation should not be delayed once it is deemed necessary. Mechanical ventilation in status asthmaticus supports gas exchange and unloads ventilatory muscles until aggressive medical treatment improves the functional status of the patient. Patients intubated and mechanically ventilated should be appropriately sedated, but paralytic agents should be avoided. Permissive hypercapnia, increase in expiratory time, and promotion of patient-ventilator synchronism are the mainstay in mechanical ventilation of status asthmaticus. Close monitoring of the patient's condition is necessary to obviate complications and to identify the appropriate time for weaning. Finally, after successful treatment and prior to discharge, a careful strategy for prevention of subsequent asthma attacks is imperative. PMID- 11940265 TI - Clinical review: non-antibiotic strategies for preventing ventilator-associated pneumonia. AB - Prevention of nosocomial pneumonia (NP) is the most important step towards reducing hospitalisation costs. The non-antibiotic prevention strategies include measures related to the correct care of the artificial airway, strategies related directly to the maintenance of the mechanical ventilator and the equipment, strategies focused in the gastrointestinal tract, and strategies related to the position of the intubated patients. While simple methods should be part of routine practice, the use of more invasive and expensive preventive measures should be used only in patients who are at high risk of NP. The appropriate use of these techniques can reduce the incidence of NP in intensive care unit patients. PMID- 11940266 TI - Clinical review: hemodynamic monitoring in the intensive care unit. AB - Since the beginning of modern anesthesia, in 1846, the anesthetist has relied on his natural senses to monitor the patient, aided more recently by simple technical devices such as the stethoscope. There has been a tremendous increase in the availability of monitoring devices in the past 30 years. Modern technology has provided a large number of sophisticated monitors and therapeutic instruments, particularly in the past decade. Most of these techniques have enhanced our understanding of the mechanism of the patients' decompensation and have helped to guide appropriate therapeutic interventions. As surgery and critical care medicine have developed rapidly, patient monitoring capability has become increasingly complex. The most important aspect in monitoring the critically ill patient is the detection of life-threatening derangements of vital functions. Aggressive marketing strategies have been promoted to monitor almost every aspect of the patient's status. However, these strategies are only telling us what is possible; they do not tell us whether they enhance patient safety, improve our therapy, or even improve patient outcome. PMID- 11940268 TI - Statistics review 1: presenting and summarising data. AB - The present review is the first in an ongoing guide to medical statistics, using specific examples from intensive care. The first step in any analysis is to describe and summarize the data. As well as becoming familiar with the data, this is also an opportunity to look for unusually high or low values (outliers), to check the assumptions required for statistical tests, and to decide the best way to categorize the data if this is necessary. In addition to tables and graphs, summary values are a convenient way to summarize large amounts of information. This review introduces some of these measures. It describes and gives examples of qualitative data (unordered and ordered) and quantitative data (discrete and continuous); how these types of data can be represented figuratively; the two important features of a quantitative dataset (location and variability); the measures of location (mean, median and mode); the measures of variability (range, interquartile range, standard deviation and variance); common distributions of clinical data; and simple transformations of positively skewed data. PMID- 11940267 TI - Bench-to-bedside review: genetic influences on meningococcal disease. AB - This review discusses the possible involvement of a variety of genetic polymorphisms on the course of meningococcal disease. It has been shown that several common genetic polymorphisms can either influence the susceptibility to meningococcal disease or can account for a higher mortality rate in patients. Gene polymorphisms concerning antibody receptors, lipopolysaccharide (LPS) binding receptors or proteins, innate complement proteins as well as cytokines and hemostatic proteins are described. The study of genetic polymorphisms might provide important insights in the pathogenesis of meningococcal disease and could make it possible to identify individuals who are at risk of either contracting or dying from meningococcal disease. PMID- 11940269 TI - Use of a rapid arterial blood gas analyzer to estimate blood hemoglobin concentration among critically ill adults. AB - OBJECTIVE: To evaluate whether measurement of the hemoglobin (Hb) concentration with a blood gas analyzer approximates that determined by a conventional coulter counter in critically ill adults. DESIGN: Prospective patient series. SETTING: A 32-bed cardiovascular, neurosurgical, trauma and medical-surgical intensive care unit in a single Canadian center. PATIENTS: We consecutively recruited 202 critically ill adults, the majority of whom had recent cardiac or vascular surgery, neurosurgery or trauma. MEASUREMENTS: The nurse obtained a single arterial blood sample within a few hours of the patient's admission to the intensive care unit. The Hb concentration was determined from each blood sample in a masked fashion, using both a blood gas analyzer and a conventional laboratory coulter counter. MAIN RESULTS: A total of 202 consecutive paired analyses were conducted. There was a highly significant correlation between the coulter counter and blood gas analyzer methods of Hb measurement (r2 = 0.98, 95% confidence interval [CI] = 0.97-0.99; P < 0.0001). Using the method of Bland and Altman, the overall mean difference in Hb concentration between the coulter counter and the blood gas analyzer was -4.3 g/l (95% CI = -11.0 to 2.4). Of the 11 (5.4%) Hb measurements that extended beyond the upper and lower 95% CI, 10 (5.0%) were within +/- 3 g/l of these confidence limits. CONCLUSIONS: An arterial blood gas analyzer may provide a valid alternative method to the traditional coulter counter for the rapid assessment of Hb concentration among critically ill adults. Since issues related to its safety, quality control, data entry and cost savings have yet to be addressed, however, use of such point of care testing should be viewed as a supplement to conventional laboratory testing. PMID- 11940270 TI - Assessment of tissue oxygen tension: comparison of dynamic fluorescence quenching and polarographic electrode technique. AB - INTRODUCTION AND METHODS: Dynamic fluorescence quenching is a technique that may overcome some of the limitations associated with measurement of tissue partial oxygen tension (PO2). We compared this technique with a polarographic Eppendorf needle electrode method using a saline tonometer in which the PO2 could be controlled. We also tested the fluorescence quenching system in a rodent model of skeletal muscle ischemiahypoxia. RESULTS: Both systems measured PO2 accurately in the tonometer, and there was excellent correlation between them (r(2) = 0.99). The polarographic system exhibited proportional bias that was not evident with the fluorescence method. In vivo, the fluorescence quenching technique provided a readily recordable signal that varied as expected. DISCUSSION: Measurement of tissue PO2 using fluorescence quenching is at least as accurate as measurement using the Eppendorf needle electrode in vitro, and may prove useful in vivo for assessment of tissue oxygenation. PMID- 11940272 TI - Concerns about the effectiveness of critical incident stress debriefing in ameliorating stress reactions. PMID- 11940271 TI - Effects of intravenous furosemide on mucociliary transport and rheological properties of patients under mechanical ventilation. AB - The use of intravenous (IV) furosemide is common practice in patients under mechanical ventilation (MV), but its effects on respiratory mucus are largely unknown. Furosemide can affect respiratory mucus either directly through inhibition of the NaK(Cl)2 co-transporter on the basolateral surface of airway epithelium or indirectly through increased diuresis and dehydration. We investigated the physical properties and transportability of respiratory mucus obtained from 26 patients under MV distributed in two groups, furosemide (n = 12) and control (n = 14). Mucus collection was done at 0, 1, 2, 3 and 4 hours. The rheological properties of mucus were studied with a microrheometer, and in vitro mucociliary transport (MCT) (frog palate), contact angle (CA) and cough clearance (CC) (simulated cough machine) were measured. After the administration of furosemide, MCT decreased by 17 +/- 19%, 24 +/- 11%, 18 +/- 16% and 18 +/- 13% at 1, 2, 3 and 4 hours respectively, P < 0.001 compared with control. In contrast, no significant changes were observed in the control group. The remaining parameters did not change significantly in either group. Our results support the hypothesis that IV furosemide might acutely impair MCT in patients under MV. PMID- 11940273 TI - Eighth World Congress of Intensive and Critical Care Medicine, 28 October-1 November 2001, Sydney, Australia: Harm minimization and effective risk management. AB - The 8th World Congress saw the presentation of several late-breaking findings, such as the role of insulin in reducing mortality, and technologies such as vital microscopy. There were heated debates for and against the role of gastric tonometry, enteral nutrition, extracorporeal membrane oxygenation, the question of 'closed' or 'open' intensive care units, and several others. The overall message was the need to study outcomes and practise intensive care in a sensitive and humane fashion. PMID- 11940275 TI - A gender-specific HIV/STD risk reduction intervention for women in a health care setting: short- and long-term results of a randomized clinical trial. AB - This study assessed the short- and long-term effect of a gender-specific group intervention for women on unsafe sexual encounters and strategies for protection against HIV/STD infection. Family planning clients (N = 360) from a high HIV seroprevalence area in New York City were randomized to an eight-session, a four session or a control condition and followed at one, six and 12 months post intervention. Using an intention-to-treat analysis, women who were assigned to the eight-session group had about twice the odds of reporting decreased or no unprotected vaginal and anal intercourse compared to controls at one month (OR = 1.93, 95% confidence interval [CI] = 1.07, 3.48, p = 0.03) and at 12-month follow up (OR = 1.65, 95% CI = 0.94, 2.90, p = 0.08). Relative to controls, women assigned to the eight-session condition reported during the previous month approximately three-and-a-half (p = 0.09) and five (p < 0.01) fewer unprotected sex occasions at one- and 12-month follow-up, respectively. Women in the eight session group also reduced the number of sex occasions at both follow-ups, and had a greater odds of first time use of an alternative protective strategy (refusal, outercourse, mutual testing) at one-month follow-up. Results for the four-session group were in the expected direction but overall were inconclusive. Thus, gender-specific interventions of sufficient intensity can promote short- and long-term sexual risk reduction among women in a family planning setting. PMID- 11940276 TI - Meta-analysis of the effectiveness of HIV prevention interventions for women. AB - The present study is a meta-analysis of the effectiveness of HIV prevention interventions for women in the USA. Twenty-four articles from 1989-1997 were included. We evaluated five ethnic groupings (All Ethnicities Combined, African American, White, Hispanic and a Mixed Ethnicity group) over four time periods (post-test, less than two months after the intervention, 2-3 months after the intervention and 6-24 months after the intervention) on three HIV-related sexuality outcome variables (HIV/AIDS knowledge, self-efficacy and sexual risk reduction behaviour). The HIV interventions appear effective at improving knowledge about HIV/AIDS and increasing sexual risk reduction behaviours for all ethnicities examined at all follow-up periods, with one exception. The findings for self-efficacy are less consistent. The interventions were less consistently effective for African-American women, for whom significant improvements in feelings of self-efficacy were only seen six months or longer after the intervention. The present analysis elucidates ethnic differences which may have previously been obscured while demonstrating convincingly that HIV interventions are generally effective for women of many different ethnicities. PMID- 11940277 TI - Factors influencing intent to get pregnant in HIV-infected women living in the southern USA. AB - This descriptive study sought to identify factors that influence HIV-infected women's intent to get pregnant. Interviews were conducted with a convenience sample of n = 322 HIV-infected women at risk for pregnancy. Participants were predominantly African-American (84.4%), single (57.9%), and ranged in age from 17 to 48 years. Forty per cent (n = 128) of the women had been pregnant since becoming HIV-positive. Potential factors influencing intent to get pregnant that were examined included demographic characteristics, HIV-related factors and personal beliefs and attitudes. In simple logistic regression models, younger age, increased motivation for child bearing, decreased perceived threat of HIV, decreased HIV symptomatology, higher traditional gender role orientation, and greater avoidance coping were all associated with greater intent to get pregnant. Following a model selection procedure, motivation for child bearing (OR = 16.05, 95% CI 7.95, 30.41) and traditional sex roles (OR = 4.49, 95% CI 1.44, 13.55) were significantly associated with greater intent to get pregnant. Traditional gender role orientation and motivation for childbearing are significant factors in predicting intent to get pregnant among HIV-infected women. These factors, as well as other non HIV-related factors, need to be routinely assessed by health care providers in developing plans of care for HIV-infected women. PMID- 11940278 TI - Women in contact with Sydney's gay and lesbian community: sexual identity, practice and HIV risks. AB - Total rates are low, but sex with a man remains the main risk for HIV transmission to women in Australia. In February 1998, 774 women in contact with gay, bisexual and lesbian communities completed a two-page questionnaire. Almost all respondents had some social contact with gay men. Two-thirds (503) thought of themselves as lesbian/dyke/homosexual/gay, 13% (100) as bisexual and 17% (133) as heterosexual/straight. In total, 212 women (27%) said they had ever had sex with a gay or bisexual man; 51 women (7%) had done so in the past six months. About 2% of the lesbians said they had recently had sex with a gay/bi man, as had 8% of the heterosexuals and 25% of the bisexuals; 25 women had recently had unprotected vaginal or anal intercourse with a male gay/bi partner. Forty-eight women (6%) had done sex work. Asked about drug use, 61 women (8%) said they had injected in the past six months. Twenty-three women had shared injecting equipment with someone, and five had shared with a gay/bi man. Compared with other women, these women have high rates of injecting drug use. If they have sex with men, these men are more likely to be gay or bisexual than are the male sexual partners of women moving in largely heterosexual milieux. For a case of so-called 'heterosexual' transmission of HIV to occur, neither person need be heterosexual. PMID- 11940279 TI - Adherence discourse among African-American women taking HAART. AB - Low adherence is the single most important challenge to controlling HIV through the use of high acting anti-retrovirals (HAART). Non-adherence poses an immediate threat to individuals who develop resistant forms of the virus as well as a public health threat if those individuals pass on treatment-resistant forms of the virus. To understand the concerns and perceptions that promote or deter adherence to antiretroviral medication by HIV-positive African-American women, we conducted in-depth interviews with 15 African-American women taking HAART. We focused on the discourse and narratives women use in talking about their adherence practice. Discourse analysis was utilized to identify and explore the sources of influence used by these women in describing their adherence practice. Roughly a third of the sample fell into each of the three self-assessed adherence categories: always adherent, mostly adherent and somewhat adherent. Among the 'always adherent', 80% of the sources of influence cited supported adherence, while only 48% and 47% of the authoritative sources cited by women in the 'mostly' and 'somewhat' categories supported adherence. Each self-assessed adherence group was characterized by its own distinctive discourse style. Findings suggest that adherence to HAART among African-American HIV-positive women would be improved by identifying those influences undermining adherence. Focused study of the 'always adherent' types is recommended. PMID- 11940280 TI - A study on female sex workers in southern China (Shenzhen): HIV-related knowledge, condom use and STD history. AB - The present study aims to investigate the perspective relations between HIV/AIDS and condom-related knowledge, condom use, history of sexually transmitted diseases (STDs) and predictive factors of condom use by female sex workers (FSW) who were sent to the Women Re-education Center (WRC) in Shenzhen, People's Republic of China. Seven hundred and one FSW were interviewed. Whereas respondents had attained certain accurate knowledge about the HIV transmission routes, misconceptions were still commonly reported. Level of condom related knowledge was not high. The prevalence of using condoms with clients was relatively low-15%- 22% reported not using condoms consistently. One of the major obstacles was 'Refusal by clients'. Significant predictive factors associated with consistency of condom use with clients were age, educational background, average number of client intakes per day, capacity in determining the use of condoms and past STD history. The respondents may have developed a false sense of safety by subjectively assessing whether their clients had a STD but they were unaware that HIV carriers may show no obvious symptoms at all. The data has confirmed the worries regarding the potential 'bridging effect' of HIV transmission as resulted by cross-border commercial sex-networking activities. PMID- 11940281 TI - Assessing quality of life in women living with HIV infection. AB - The purpose of this study was to examine the impact of demographic, clinical and psychosocial variables on health-related quality of life in women living with human immunodeficiency virus infection. Subjects included 82 women 18 years of age or older who were currently seeking treatment at an outpatient clinic. The Medical Outcomes Study-Short Form 20 (SF-20) assessment consisted of patient self ratings on 20 items measuring six distinct health dimensions. Additional data were collected on age, ethnicity, education, employment status, length of time since HIV diagnosis, CDC stage of disease, total CD4 count, number of symptoms, Karnofsky Performance Scores, social support and discrimination. Multiple regression analyses were performed using each of the six health dimension scores as dependent variables and a combination of ten demographic, clinical and psychosocial variables as independent variables. All of the six multiple regression equations were statistically significant, with R(2) values ranging from 0.35 to 0.44. Age, Karnofsky Performance Score and social support were statistically significantly related to several scale scores (p < 0.05). These results provide baseline data on clinical, demographic and psychosocial determinants of quality of life in a female patient population. Quality of life information can assist programme planners in developing social and medical service delivery for women in the community thus facilitating the development of treatments, programmes and interventions aimed at increasing quality of life for this population. PMID- 11940283 TI - The 'family' context of HIV: a need for comprehensive health and social policies. AB - This paper reports on the findings from a multi-site psychosocial study of Canadian families with HIV-positive mothers. A total of 110 adults, representing 91 families across Canada participated in interviews. Qualitative analysis revealed a number of themes including: a complex web of personal, health and family concerns; the needs of children; family finances; disclosure dilemmas; and social experiences and challenges. These themes reflect an intricate and dynamic picture of parental and family life for adults and children living with HIV infection. Nowhere in the literature do we see HIV framed as a 'family infection'. Surveillance reporting reflects information on infected adults and children but not family groupings. Yet with HIV several family members and multiple generations as well as single or both parents may be infected, highlighting the importance of 'family HIV' as a framework for health policy and programme development. At issue is the problem that medical and other institutions view issues of surveillance, treatment and care through the lens of the infected individual, rather than being family focused. Often it is only in the context of identifying support, or barriers to support, for the medically diagnosed individual that biological or socially created families become a focus of concern. The failure to situate both chronic and life-threatening illnesses within the family setting has serious quality of life and planning consequences for parents and children living with HIV infection as well as other illnesses. PMID- 11940282 TI - Redefining treatment: how biological mothers manage their children's treatment for perinatally acquired HIV. AB - Current medical thinking supports early initiation of aggressive approaches to the management of perinatally acquired HIV infection. Biological mothers, however, may not endorse this way of managing their child's condition. For this study, grounded theory methods were used to conduct secondary analysis of eight semi-structured in-depth interviews with parents of children with perinatally acquired HIV. Biological mothers' perspectives on their children's treatment for perinatally acquired HIV infection, the strategies they used to manage their children's treatment and sociocultural influences on mothers' beliefs and actions are explored and described. Creation of a framework that may be useful for future research and recommendations for clinical practice are offered. PMID- 11940284 TI - Psychosocial intervention in the process of empowering families with children living with HIV/AIDS-a descriptive study. AB - The notion of 'family empowerment' is of crucial importance in any kind of psychosocial intervention directed to families with children living with HIV/AIDS. The goal is to restore self-efficacy and self-esteem, encourage positive coping and help family members make their own choices. Two settings for psychosocial intervention are described: an outpatient clinic within a larger medical institution and a therapeutic camp for families with children. Different psychotherapeutic and counselling strategies and approaches are briefly presented. Preliminary results based on work with families with children infected and/or affected by HIV/AIDS are described. The conclusion is that interventions that involve different settings seem to be most effective, especially for the most vulnerable families. The important issue is also collaboration between different service providers and NGOs. PMID- 11940285 TI - Tell them you love them because you never know when things could change: voices of adolescents living with HIV-positive mothers. AB - This study analyzed the qualitative and quantitative responses of 60 minority teens affected by HIV. Relationships between adolescents' reports of: (1) feeling different, (2) having secrets, (3) worrying, and (4) caretaking were revealed. Six themes emerged from the content analysis of qualitative data that reflect how these adolescents have been affected by HIV: (1) core assumptions about life, (2) fearful anticipation of the death, (3) stigma and isolation, (4) current and future losses, (5) family role reassignment, and (6) lack of resources. Group treatment is presented as an effective method of intervention for adolescents living with an HIV-positive mother. PMID- 11940288 TI - [Multicenter clinical trial on the efficacy of thrombolytic therapy with urokinase and/or anticoagulant with low molecular weight heparin in acute pulmonary embolism]. AB - OBJECTIVE: To evaluate the efficacy of thrombolytic therapy with urokinase and/or anticoagulant with low molecular weight heparin in patients with acute pulmonary embolism. METHODS: 127 patients with documented acute pulmonary embolism were treated with different regimens: thrombolysis combined with anticoagulant therapy in 70 cases, thrombolytic therapy alone in 31 cases and pure anticoagulant therapy in 26 cases. RESULTS: Dyspnea was the most common symptom (91.3%), then cough (74.0%), chest pain (63.0%) and syncope (21.3%). As compared with pre treatment in each group, there was significant improvement in respiratory rate, heart rate, partial pressure of oxygen in artery and partial pressure of carbon dioxide in artery. The effective rate of thrombolysis combined with anticoagulant therapy, thrombolytic therapy and anticoagulant therapy were 90.0%, 77.4% and 61.5%, respectively, the total effective rate of thrombolytic therapy was 86.1%. The best efficacy was demonstrated in patients within one week after onset of symptoms. The therapy might be effective for cases with duration longer than two weeks after the onset of symptoms. CONCLUSION: Thrombolytic therapy with urokinase combined with low molecular weight heparin is effective and safe for acute pulmonary embolism. PMID- 11940289 TI - [Microsomal epoxide hydrolase gene polymorphism and susceptibility to chronic obstructive pulmonary disease in Han nationality of North China]. AB - OBJECTIVE: We investigated whether polymorphism in gene for microsomal epoxide hydrolase (mEH) has any bearing on individual susceptibility to the development of chronic obstructive pulmonary disease. METHOD: The genotypes of 55 patients with COPD and 52 healthy smoking control subjects were tested with polymerase chain reaction followed by restriction fragment length polymorphism for mEH gene. RESULT: The frequency of polymorphic genotypes of mEH showed no difference between the COPD group and the control group. In COPD group mEH exon 3 homozygous wild-type, heterozygote and homozygous mutant was 27.3%, 27.3% and 45.5% respectively and exon 4 homozygous wild-type, heterozygote and homozygous mutant was 72.7%, 18.2% and 9.1% respectively. CONCLUSION: Genetic polymorphism in mEH is not associated with development of COPD in Han nationality of North China. PMID- 11940290 TI - [The optimal pressure of sustained inflation for alveolar recruitment in acute respiratory distress syndrome rabbit model]. AB - OBJECTIVE: To determine the optimal pressure of sustained inflation (SI) in treatment of acute respiratory distress syndrome (ARDS). METHODS: SI was applied at pressures of 1 approximately 6 times of mean airway pressure (P(m)) for 20 s on saline alveoli-lavaged anesthetized rabbits. Hemodynamics, pulmonary mechanics and gas exchange were observed before, during, and 2 min, 5 min after applying SI. Lung histology was observed after the experiment. RESULTS: When the pressure of SI was higher than 3P(m), partial pressure of arterial oxygen (PaO(2)) and arterial oxygen saturation were improved. The difference of PaO(2) before and during SI were (65 +/- 40) mm Hg and (52 +/- 25) mm Hg respectively in the 5P(m) and 6P(m) group, being significantly higher than that in the 1P(m) group [(-5 +/- 4) mm Hg, P < 0.05]. After SI, dynamic pulmonary compliance (C(dyn)) in the 5P(m) group [(3.06 +/- 0.24) ml/cm H(2)O] was increased markedly, as compared with C(dyn) before SI [(1.09 +/- 0.18) ml/cm H(2)O, P < 0.05]. SI with 5P(m) resulted in immediate significant increase in lung volume [(44.0 +/- 3.1) ml/kg in the 5P(m) group, (8.3 +/- 0.7) ml/kg in the 1P(m) group]. Histologically, Smith lung injury score was 4.40 +/- 1.66 in the 5P(m) group, which was less than that in the group of ARDS model 6.10 +/- 0.77. SI with 6P(m) led to alveolar overdistention. With the increasing of SI pressure, mean arterial pressure decreased markedly. CONCLUSION: 5P(m) (25 approximately 35 cm H(2)O) may be the optimal pressure of SI in rabbits with severe ARDS. PMID- 11940291 TI - [Comparison of the clinical efficacy of recombinant tissue-type plasminogen activator thrombolysis with primary coronary stenting in acute myocardial infarction]. AB - OBJECTIVE: To compare the efficacy of low dose recombinant tissue-type plasminogen activator (rt-PA) thrombolysis with primary coronary stenting after acute myocardial infarction (AMI). METHODS: Of 261 patients with first AMI, 131 patients were performed with low dose rt-PA intravenously, and 130 patients with primary coronary stenting. The clinical efficacy of these two groups was compared. RESULTS: The age, time from onset of chest pain to hospital presentation and infarct location between these two groups were comparable. The patency rate of infarct-related artery (IRA) in patients with thrombolysis group was significantly lower than that of patients with primary stenting group (P < 0.00001). The recurrent myocardial infarction, selective coronary stenting of patients with thrombolytic therapy were higher than that of patients with primary stenting group (7.6% vs 1.5%, P < 0.05; 20.6% vs 0, P < 0.001, respectively). The LVEF in patients with thrombolysis group was lower than that of stenting group [(55.6 +/- 13.4)% vs (65.8 +/- 9.2)%, P < 0.0001]. Total hospitalization time of thrombolysis group was longer than that of stenting group [(16 +/- 7) days vs (11 +/- 4) days, P < 0.0001]. The mortality of thrombolysis group was higher than that of stenting group, but this difference was not significant (6.1% vs 3.1%, P > 0.05). CONCLUSIONS: Compared with low dose rt-PA thrombolytic therapy after AMI, primary coronary stenting has a higher patency rate of the IRA, better cardiac function and shorter hospitalization time. PMID- 11940292 TI - [High incidence of renal artery stenosis in patients undergoing coronary angiography]. AB - OBJECTIVE: It is important to identify patients at risk for atherosclerotic renal artery stenosis because renal artery stenosis is a progressive disease and a potentially correctable problem. This study is to evaluate the prevalence of angiographically-determined renal artery stenosis in a patient population undergoing routine coronary angiography, and to identify the risk factors for renal artery stenosis. METHODS: Before the procedure, demographic data, medical history, physical findings and laboratory data were obtained. After coronary angiography, abdominal aortography was performed to screen for the presence of renal artery stenosis over a 8-month period. In 370 patients (237 males, 133 females), mean age was 60.6 (29 to 81) years. And an univariate and multivariate Logistic regression analysis was made to explore the association of the clinical variables with renal stenosis. RESULTS: Renal artery narrowing was identified in 106 (28.6%) patients. Significant renal artery stenosis, defined as a narrowing of the diameter by more than 50%, was found in 63 patients (17.0%) and insignificant (< 50% diameter narrowing) stenosis was found in 43 patients (11.6%). Significant unilateral stenosis was present in 41 (11.1%) patients and bilateral stenosis was present in 22 (5.9%) patients. By univariate and multivariate logistic regression analysis, the association of clinical variables with renal artery stenosis was assessed. Multivariable predictors included age (OR = 1.06, 95% CI: 1.03 approximately 1.10, P < 0.0001), severity of coronary artery disease (OR = 1.65, 95% CI: 1.34 approximately 2.03, P < 0.0001) and peripheral vascular disease (OR = 2.64, 95% CI: 1.27 approximately 5.47, P = 0.0095). The variables such as sex, body mass index, hypertension, smoking history, hyperlipidemia, myocardial infarction, congestive heart failure, renal insufficiency and diabetes mellitus were not associated. CONCLUSION: Renal artery stenosis is a frequent finding in patients undergoing coronary angiography, especially in older people with generalized atherosclerosis. PMID- 11940293 TI - [The clinical features and tumor cells characteristics of splenic marginal zone lymphoma]. AB - OBJECTIVE: To deepen the understanding of splenic marginal zone lymphoma (SMZL) and improve the level of diagnosis and therapy. METHODS: A typical case of SMZL, a 61 year old female with lymphocytosis and splenomegaly found fortuitously, was reported. The pathologic, immunologic and genetic features of tumor cells in peripheral blood, bone marrow and spleen were studied with light microscopy, phase contrast microscopy, scanning electron microscopy, immunohistochemical method, flow cytometry, G chromosome banding technique and PCR for studying the pattern of IgH gene rearrangement. RESULTS: The spleen was large with uniform parenchyma and smooth surface. There were multiple small gray-white nodules on sections. Histologically, the neoplastic cells replaced the marginal and mantle zones with complete replacement of germinal centers in the white pulp. The neoplastic cells were predominantly of small to medium size with oval or slightly irregular nuclei. Lymph nodes in the splenic hilum were infiltrated by tumor cells. Immunophenotypic analysis demonstrated that the lymphocytes in the bone marrow expressed CD(20), HLA-DR, CD(45) RA and bcl-2. The monoclonal pattern of IgH gene rearrangement in peripheral blood and bone marrow was found to be the same as that in spleen. After splenectomy, COP chemotherapy and IFNalpha-2a were given and the abnormally increased lymphocytes decreased to normal level. Seven months later the monoclonal rearranged immunoglobulin heavy chain gene pattern changed to polyclonal pattern. CONCLUSION: Splenomegaly, lymphocytosis in peripheral blood and bone marrow without lymph node enlargement and leukocytosis are clinical characters of SMZL. Presence of monoclonal rearranged IgH gene is in favor of the diagnosis. Splenectomy should be done earlier in suspicious patients to avoid malignant transformation. PMID- 11940295 TI - [The impact of plasma homocysteine level on development of retinopathy in type 2 diabetes mellitus]. AB - OBJECTIVE: An increased plasma homocysteine level is an important risk factor for vascular disease in the general population. However, the role of hyperhomocysteinemia in the development of type 2 diabetic retinopathy (DR) is still unknown. Therefore, our aim was to determine the relationship between the fasting plasma homocysteine (Hcy) levels and the presence of diabetic retinopathy in patients with type 2 diabetes. METHODS: The study group consisted of 55 cases of type 2 diabetic patients (DM); 19 healthy persons served as control (CON, 12 men and 7 women). The DM group were subdivided into a group without diabetic complications (NDC, 39 cases, 17 men and 22 women) and a group with DR (16 cases, 8 men and 8 women). The renal function and the ratio of albumin/creatinine (Alb/Cr) were within normal range in all the subjects. The presence and the grade of retinopathy were determined by an ophthalmologist. Plasma total Hcy (tHcy) was measured by automated high-performance liquid chromatography (HPLC) with reverse phase separation and fluorescence detection. RESULTS: Plasma tHcy levels were significantly different among DR, NDC and CON groups (F = 2.405, P = 0.031). Plasma total Hcy was significantly higher in DR group [(14.7 +/- 5.28) micromol/L] than in NDC [(11.3 +/- 4.94) micromol/L] and CON group [(9.65 +/- 2.66) micromol/L]. There was no difference between NDC group and CON group. In DR group, plasma tHcy was significantly higher in proliferative diabetic retinopathy (PDR) group than that in background diabetic retinopathy (BDR) group (t = 2.405, P = 0.031). Hyperhomocysteinemia was defined as tHcy level > 14.97 micromol/L and was seen in 4 patients in PDR group, the rate being significantly higher than that in BDR group. CONCLUSION: It is suggested that hyperhomocysteinemia may be a risk factor for the development and progression of type 2 diabetic retinopathy. PMID- 11940294 TI - [The change in sensitivity of methicillin-resistant Staphylococcus aureus to vancomycin]. AB - OBJECTIVE: To determine the shift of susceptibility of methicillin-resistant Staphylococcus aureus (MRSA) isolates to vancomycin in recent years. METHODS: Antimicrobial susceptibility was determined by disk diffusion test (K-B method). The t-test was used to compare the mean diameters of zone of inhibition. RESULTS: The clinical isolates of MRSA from 1994 to 2000 were 218, 187, 292, 237, 351, 886 and 652 in number respectively. The mean diameters (mm) of the inhibition zone of MRSA isolates each year were 19.9, 19.1, 18.6, 17.4, 18.4, 18.2 and 17.8. Comparison of the diameters of the inhibition zone of MRSA isolates (K-B method) showed that the mean diameter of the inhibition zone from the 652 MRSA isolates in 2000 was much less than that of the 218 MRSA isolates in 1994 and the difference was significant (P < 0.001). The resistant rates to chloramphenicol, ciprofloxacin, erythromycin, gentamicin, tetracycline and trimethoprim/sulfamethoxazole were 44.0%, 73.7%, 89.1%, 65.7%, 61.0% and 67.0%. CONCLUSION: Our observation demonstrated that the susceptibility of MRSA to vancomycin gradually decreased from 1994 to 2000. It is important to monitor the antimicrobial susceptibility of Staphylococcus aureus to vancomycin. PMID- 11940296 TI - [The expression and implication of proliferating cell nuclear antigen and p53 protein in reflux esophagitis lesions]. AB - OBJECTIVE: To characterize expression of proliferating cell nuclear antigen (PCNA) and p53 protein in esophageal mucosa of reflux esophagitis (RE) and which returning to normal endoscopically after intervening therapy. METHODS: Esophageal biopsies were taken from 213 case of RE and 26 case returning to normal endoscopically after intervening therapy, which were examined by pathology and avidin-biotin-peroxidase complex (ABC) method. RESULTS: The number of PCNA positive cells in esophageal mucosa increased from normal to RE and to dysplasia, with the values being (192 +/- 120)/mm(2), (326 +/- 200)/mm(2), (596 +/- 208)/mm(2) respectively. There was statistically significant difference between intergroups (P < 0.01). The positive rates of p53 protein in esophageal mucosa increased from normal to RE and to dysplasia and they were 5.9%, 34.4%, 57.6%. Marked difference was found between intergroups (P < 0.01). The number of PCNA positive cells in esophageal mucosa of RE patients which appear to be normal under endoscope after intervening therapy [(306 +/- 214)/mm(2)] and the positive rates of p53 protein (26.9%) were significantly lower than those in RE patients before intervening therapy (P < 0.01), but were significantly higher than those in normal controls (P < 0.01). CONCLUSIONS: The expression of PCNA and p53 protein increased with the severity of esophageal reflux lesions. The case of reflux esophagitis who returned to normal endoscopically after intervening therapy need keeping on therapy and being followed by endoscopy, because their esophageal mucosa didn't become normal at the molecular level. PMID- 11940297 TI - [Heparin blocks the binding of advanced glycation end products to its receptors on human monocytes]. AB - OBJECTIVE: Previous studies have demonstrated the accumulation of advanced glycation end products (AGE) in patients with chronic renal failure (CRF), but the mechanism is not completely understood. This study was performed to elucidate the effect of heparin on binding of AGE to its receptors on human monocytes. METHOD: Human serum albumin (HSA) modified with AGE was prepared in vitro and was radioiodinated with carrier-free [(125)I]. Human peripheral blood monocytes obtained from dialysis patients and normal volunteers were isolated by Ficoll hypaque centrifugation technique. Specific binding (125)I-AGE-HSA to the AGE receptor on human monocytes was measured by radioactive ligand-receptor binding assay. RESULT: The binding of (125)I-AGE-HSA to its receptors in dialysis patients was inhibited by 27% 15 minutes after starting of hemodialysis using heparin as anti-coagulant. These effects continued 6 hours and resume to the levels of predialysis after 24 hours. There was no differences in binding of (125)I-AGE-HSA to monocytes between pre-and post-dialysis session when low molecule weight heparin (LMWH) was used as the anti-coagulant. In vitro study further demonstrated that exposure of normal monocytes to heparin-containing media inhibited the binding of (125)I-AGE-HSA to its receptor with a dose dependent manner. LMWH did not inhibit such binding. CONCLUSION: Heparin blocks the binding of AGE to its receptors on monocytes and may therefore interrupt the clearance and degradation of AGE. This may be one of the mechanisms by which AGE accumulation occurs in patients with hemodialysis. PMID- 11940298 TI - [Expression of matrix metalloproteinase-9 and tissue inhibitor of metalloproteinase-1 in IgA nephropathy]. AB - OBJECTIVE: To explore the expression of matrix metalloproteinase-9 (MMP-9) and tissue inhibitor of metalloproteinase-1 (TIMP-1) in the renal tissue of patients with IgA nephropathy. METHODS: Immunohistochemistry and in situ hybridization assays were applied to detect the expression of mRNA and protein of TIMP-1, as well as the protein expression of MMP-9 in renal tissue of 38 patients with IgA nephropathy. RESULTS: MMP-9 was normally expressed, at a low level, in the glomerular endothelial cells and visceral epithelial cells, as well as in tubular epithelial cells and walls of vessels. The expression of MMP-9 was significantly increased in mesangial proliferative glomeruli and interstitial vascular walls of IgA nephropathy patients (P < 0.001), but markedly decreased in sclerotic glomeruli and not apparently altered in the tubule. No expression of TIMP-1 could be detected in normal renal tissue. TIMP-1 was only slightly expressed in the mesangioproliferative glomeruli of IgA nephropathy patients. TIMP-1 expression was increased notably in partially sclerotic glomeruli, and most prominently expressed in tubulointerstitium (P < 0.01), mainly in tubular epithelial cells, interstitial cells, and vascular endothelial cells. The expression of TIMP-1 was distinctly associated with the level of serum creatinine (P < 0.05), tubulointerstitial fibrosis (P < 0.01), and tubulointerstitial infiltration of leukocytes (P < 0.01). MMP-9 expression did not correlate with proteinuria, but negatively correlated with serum creatinine levels (P < 0.05). CONCLUSION: The abnormal expressions of MMP-9 and TIMP-1 may contribute to the progression of IgA nephropathy. PMID- 11940299 TI - [The effect of exercise during hemodialysis on adequacy of dialysis]. AB - OBJECTIVE: To investigate the effect of exercise on dialysis adequacy during hemodialysis (HD). METHODS: 20 studies were performed in 20 maintenance hemodialysis patients. Each study consisted of a pair of HD treatment; the blood and dialysate flow rates, dialyser and treating time were the same. An adapted exercise cycle was attached to the dialysis bed. The patients of exercise group pedaled the exercise cycle throughout the dialysis, and had a 5 - 10 minutes rest if they needed. while the patients of a control group had no exercise. BUN, Scr and UA were measured just before and after dialysis immediately, and 60 minutes after dialysis. Furthermore, KT/V, SRI, AUR and nPCR were detected by on-line dialysis monitor. RESULTS: In the exercise group, after HD, both BUN and Scr decrease rates were significantly higher, while the rebound of BUN, Scr and UA were significantly lower than those in the control group (P < 0.05 approximately 0.01). KT/V, AUR and SRI were significant higher in the exercise group than those in the control group (P < 0.05 approximately 0.01 respectively), while nPCR showed no significant difference between the two groups. CONCLUSION: Exercise during HD is a practicable new way to improve the efficiency of dialysis. PMID- 11940300 TI - [Preliminary results of stenting revascularization as treatment for renal artery stenosis]. AB - OBJECTIVES: To evaluate the safety and short-term efficacy of stent revascularization as treatment for renal artery stenosis. METHODS: Percutaneous transluminal renal angioplasty with stent (PTRAS) was performed on 30 consecutive patients with severe renal artery stenosis for poorly controlled hypertension or preservation of renal function. They subsequently underwent 3-month clinical follow-up for the effect of the procedure on renal function, blood pressure control and the number of antihypertensive medications used. RESULTS: Angiographic success was obtained in 29 (96.7%) of the 30 patients after PTRAS. 3 months after the procedure systolic and diastolic blood pressures significantly decreased (from 173.5 to 135.8 mm Hg and from 95.8 to 75.6 mm Hg, respectively; P < 0.001) and less antihypertensive medications were taken (from 2.5 to 1.5). Blood pressure in 5 (16.7%) of the 30 patients became normal without taking any antihypertensive medication and blood pressure control was more facile in 22 (73.3%) of the patients. However, there was no improvement in 3 (10.0%) of the patients. Creatinine slightly decreased in 2 (6.7%) of the 30 patients and remained stable in 28 (93.3%) of the 30 patients. There was no statistical significance in this respect. CONCLUSIONS: Renal artery stent revascularization has a short-term beneficial effect on blood pressure control and a nondeleterious effect on renal function. The long-term efficacy should be investigated. The procedure is safe in usual. PMID- 11940301 TI - [Tumor necrosis factor-alpha enhances type I inositol 1, 4, 5-triphosphate receptor expression in rat glomerular afferent arterioles smooth muscle cells]. AB - OBJECTIVE: To study the effect of tumor necrosis factor-alpha (TNF-alpha) on type I inositol 1, 4, 5-triphosphate receptor (IP(3) R) expression in rat glomerular afferent arterioles smooth muscle cells (RASMC). METHODS: Isolation and culture of RASMC and detection of type I IP(3) R protein and type I IP(3) R mRNA in RASMC after TNF-alpha treatment were carried out with Western blot and Northern blot assay. RESULTS: TNF-alpha enhanced the expression of type I IP(3) R protein and type I IP(3) R mRNA in RASMC; TNF-alpha did not influence the half life of type I IP(3) R mRNA in RASMC treated with TNF-alpha. CONCLUSION: TNF-alpha plays a role in the development of renal dysfunction. The mechanisms may be that TNF-alpha takes part in the change of signal transduction in RASMC. TNF-alpha may act on the promoter of type I IP(3) R mRNA in RASMC and results in the expression of type I IP(3) R protein that stimulates release of intracellular Ca(2+) in RASMC and induces contraction of RASMC. The renal blood flow diminution leads to the development of renal dysfunction. PMID- 11940302 TI - [Intestinal infection and irritable bowel syndrome]. AB - OBJECTIVE: To determine whether intestinal infection plays a role on the pathogenesis of irritable bowel syndrome (IBS). METHODS: 295 patients who had no previous history of functional bowel disorder had received treatment for dysentery (n = 235) or for acute bowel infection at the hospital between April October, 1998, were followed up for 1 - 2 years and evaluated for their subsequent bowel habits. A cohort study of 243 subjects using their siblings, husbands or wives who did not have dysentery or acute bowel infection at the same period was taken as control. Furthermore, the contents of mRNAs of IL-1alpha, IL 1beta and IL-1ra in the mucosa at the terminal ileum and recto-sigmoid junctions were determined and compared using RT-PCR method in 30 IBS patients and 12 controls. RESULTS: (1) Sixty-six (22.4%) patients were reported to have functional bowel disturbance, and 24 (8.1% total and 10.2% among cases of dysentery) developed IBS in the study group, whereas, only 7.4% had altered bowel habit and 0.8% had IBS in the control group (P < 0.01). (2) The risk of having functional bowel disturbance was higher in patients who suffered from a longer duration (> 8 d, OR = 3.5) of dysentery. (3) The IL-1beta mRNA level in the mucosa of terminal ileum and recto-sigmoid junction of IBS patients with dysentery was higher than that of controls and IBS patients without dysentery (P < 0.01). CONCLUSION: Intestinal infection plays a role on the pathogenesis of IBS through some immunological factors. PMID- 11940303 TI - [Pancreatic encephalopathy and Wernicke encephalopathy]. AB - OBJECTIVE AND METHOD: 3 cases of acute severe pancreatitis with symptoms of central nervous system were presented and 30 Chinese articles of acute pancreatitis (AP) with encephalopathy found in the literature were reviewed in CNKI. RESULTS: There were 185 cases of AP with encephalopathy altogether, 84% of them being severe cases, with male slightly outnumbering female and biliary pancreatitis as the predominating type. Encephalopathy appeared in the early course of AP was a part of multiple organ failure which results from acute systemic inflammatory reaction. This is, in fact, pancreatic encephalopathy (PE). There were altogether 107 cases of PE and 61 of them died (57%). Encephalopathy appeared in the later course of AP (> 2 weeks or in recovery period) was caused by deficiency of Vit B(1). This is Wernicke encephalopathy (WE). There were 78 cases of WE and 26 died (33%). The difference between the two groups was significant (P < 0.01). Supplement of thiamine in time was the reason for lower mortality in WE. Of the 78 cases with WE, 17 having supplement of Vit B(1) all survived. Among those who did not receive Vit B(1), the mortality was 43%; this was not so much different with the mortality of PE (P > 0.05). CONCLUSIONS: This article discussed in detail the pathogenesis, diagnosis and management of PE and WE. Treatment of PE includes early diagnosis and comprehensive intensive therapeutic measures in intensive care unit. Patients who have been in fasting for a long time (more than 10 days) should be given thiamine intramuscularly in case WE occurs. As soon as WE is diagnosed, a large doses of Vit B(1) should be given immediately, is the key to reduce the mortality of encephalopathy complicating AP. PMID- 11940304 TI - [Clinical and pathological features of 5 adult patients with Langerhans cell histiocytosis]. AB - OBJECTIVE: To improve the identification of Langerhans cell histiocytosis (LCH). METHODS: 5 adult cases of LCH treated in our hospital since 1990 were studied, the clinical data and pathological findings were analyzed. RESULTS: The sex ratio of male to female was 3 to 2, the mean age in this series of 5 patients was 36.8 years. All patients presented with multifocal or disseminated form of the disease. The involved organs included lung, skeleton, central nervous system, skin, liver, spleen and lymph node. The radiographic presentation of lung involvement includes diffuse micronodular densities (1 case), interstitial fibrosis with cyst formation (2 cases), limited infiltration in unilateral lung field (1 case) and isolated mass (1 case). Sharply demarcated bony rarefaction in radiogram was found in 3 cases. The presence of specific Langerhans cell in the focal lesion, which was the pathological basis for the definitive diagnosis of LCH, was detected in all patients. Systemic chemotherapy with corticosteroid and anthracycline based regimen was given to 5 patients and it caused retraction of the enlarged liver and spleen in 1 case, amelioration of lymph nodes and skin involvement in 2 cases and diminution of enlarged thyroid gland in 1 case, while it had less benefit in lung and bone involvement or to diabetes insipidus. CONCLUSIONS: LCH can be observed in young adults or middle-aged subjects. It should be suspected in patients who presented with diabetes insipidus, characteristic bony lesion and pulmonary involvement. The definitive diagnosis should be made pathologically in time. The proper treatment with low-dose radiotherapy and/or systemic chemotherapy should be taken based on the number and the function of systems involved. PMID- 11940305 TI - [The clinical significance of plasma endothelin measurement in acute organophosphorus pesticide poisoning]. AB - OBJECTIVE: To investigate the clinical significance of plasma endothelin (ET) contents in patients with acute organophosphorus pesticide poisoning (AOPP). METHODS: 69 cases of AOPP were divided into 3 groups: mild poisoning, moderate poisoning and severe poisoning groups. 25 healthy subjects served as controls. The contents of plasma ET of the patients and healthy subjects were determined. RESULTS: No significant difference was noted between the contents of plasma ET of the control and the mild group (50.85 +/- 8.93) ng/L vs (56.18 +/- 12.80) ng/L, P > 0.05. The contents of plasma ET of mild cases were only slightly increased. The contents of plasma ET in the moderate (72.61 +/- 14.21 ng/L) and serve group (110.32 +/- 17.38 ng/L) were significantly higher than those of the control group (P < 0.01, P < 0.01). The contents of plasma ET in the moderate were significantly higher than those of the mild group (P < 0.01). The contents of plasma ET in the serve group were significantly higher than those of the moderate (P < 0.01). CONCLUSION: With the increase of AOPP severity, the contents of plasma ET elevated gradually. Plasma ET may serve as an important index for the clinical course, therapeutic efficacy and prognosis of AOPP. Early regression of the contents of plasma ET may play an active role in preventing and curing multiple system organ failure resulted from AOPP and in improving the prognosis. PMID- 11940306 TI - [The effects of glucocorticoids and insulin on serum leptin level and its diurnal rhythmicity]. AB - OBJECTIVE: To investigate the effects of glucocorticoids and insulin on serum leptin level and its diurnal rhythmicity in patients with Cushing's syndrome and obesity as well as in normal subjects. METHODS: Fifteen patients with Cushing's syndrome and 10 with obesity as well as 9 normal subjects were investigated, with their serum leptin, cortisol and insulin levels measured 6 times a day (8:00, 12:00, 16:00, 20:00, 24:00, 3:00). RESULTS: The diurnal rhythmicity of serum leptin was still preserved in all the three groups, with peak appearing at 24:00 approximately 3:00 and nadir at 8:00 approximately 12:00. However, the relative diurnal excursion was negatively correlated with body mass index (BMI). The peak positive effects of insulin on leptin secretion delayed by 4 to 8 hours. CONCLUSIONS: Glucocorticoids can increase serum leptin levels, but the changes of the diurnal rhythmicity of cortisol do not affect the diurnal rhythmicity of leptin. The relative diurnal excursion of leptin is negatively correlated with body fat, i.e, the fatter subjects usually have the blunter leptin diurnal rhythmicity. Insulin exerts an effect on leptin secretion in a delayed manner, indicating its role in maintaining the diurnal rhythmicity of serum leptin. PMID- 11940307 TI - [A clinical trial using zidovudine and lamivudine plus indinavir triple therapy in Chinese individuals with human immunodeficiency virus infection]. AB - OBJECTIVE: Evaluation of high active antiretroviral therapy (HAART) in human immunodeficiency virus and acquired immunodeficiency syndrome (HIV/AIDS) patients receiving combined antiretroviral therapy in China. METHODS: We initiated the first efficacy trial of zidovudine (AZT) 600 mg/d and lamivudine (3TC) 300 mg/d (brand name: Combivir) plus indinavir (2 400 mg/d) in 15 Chinese chronically infected with HIV in May 1999 at Beijing. RESULTS: There was a rapid reduction of 2.7 log in the plasma viremia levels in 15 cases 3 months, after treatment, from a mean baseline of 90 743 copies/ml. The mean CD(4) cell counts increased by 67 cells/microliter from a baseline mean value of 471 cells/microliter and the mean CD(8) cell counts reduced by 192/microliter after 12 months of therapy. The ratio of CD(4)/CD(8) increased from 0.35 to 0.56. The average naive CD(4) cell (CD(45)RA + CD(62)L +) count and naive CD(8) cell (CD(45)RA + CD(62)L +) count increased 42 and 19/microliter respectively after one-year treatment. This drug regimen was well tolerated, with mild nausea in all, transient elevated serum bilirubin in three and kidney stone in two patients. CONCLUSION: It is effective for the virus with different genotype. The results will form a scientific foundation for the development of therapeutic strategies for HIV infection in China. PMID- 11940308 TI - [Epidemiological study of hypertension in Heilongjiang province]. AB - OBJECTIVE: To find out the epidemiological features of hypertension in Heilongjiang Province, so as to provide guidance and scientific ebidence for hypertensive prophylaxis and treatment. METHODS: >From the year of 1959 to 1999, 289 157 people in total with age >/= 15 years old (male 154 091, female 135 066) were investigated with stratified chunk method in rural and urban areas in Heilonging Province. RESULTS: Hypertensive incidence in Heilongjiang is much higher than the average level in China (P < 0.01). Hypertensive incidence has become higher and higher during the 40 years it increased 105%. It is in the recent ten years that hypertensive incidence increased most fast. Hypertensive incidence increased more obviously in male than in female (P < 0.01). Hypertensive incidence also increased with aging. Hypertensive incidence has increased both in urban and rural areas, with the urban incidence higher than rural (P < 0.01). Blood lipids (TC, TG, LDL-C, VLDL-C) of the subjects receiving this analysis in the year of 1999 was much higher than those in 1985 (P < 0.01). Alcohol drinking is related to hypertension (OR = 2.18). Smoking is also related to hypertension (OR = 1.29). People with body mass index (BMI) >/= 20 had higher hypertensive incidence (17.56%) than those with BMI < 20 (7.69%). 39 possible related factors of hypertension were analyzed; it is shown that profession of cadre, high BMI, alcohol drinking, male gender, mother's history of hypertension high salt diet and old age are the main risk factors of hypertension. CONCLUSION: The programs of prophylaxis and treatment of hypertension should be enhanced in this province. PMID- 11940309 TI - [The plasma levels of proinsulin and true insulin and the effects of ramipril in hypertension]. AB - OBJECTIVE: To explore the dynamic changes of plasma proinsulin (PI) and true insulin (TI) and their relation with essential hypertension (EH) as well as to evaluate the therapeutic effect of ramipril. METHODS: PI, TI and immunoreactive insulin (IRI) were measured in 44 cases of EH, 24 normal subjects and 21 cases of EH before and after treatment with ramipril. RESULTS: The mean plasma level of PI and IRI in EH was significantly higher than that in the control group (P < 0.01); there was no significant difference in the level of TI (P > 0.05). PI was positively related to TI, There was no significant relation between PI with SBP and DBP. Plasma concentration of IRI and PI (P < 0.05), decreased remarkably, but the level of TI remained unchanged after therapy with ramipril (n = 21). CONCLUSION: The plasma concentration of PI is elevated in patients with EH due probably to impaired function of beta-cells of the pancreatic islets. Ramipril can decrease the level of PI and improve pancreatic beta cell function. PMID- 11940311 TI - [The effects of C2 gene on growth of gastric cancer cells]. AB - OBJECTIVE: To investigate the effect of a newly cloned full length gene, C2 gene, which encodes human eukaryotic translation initiation factor, on growth of gastric cancer cells. METHODS: A constructed eukaryotic vector carrying the full length of C2 gene was amplified, purified and transfected into gastric cancer cell line-SGC 7901. Expression of C2 protein was examined with fluorescence activated cell sorting (FACS) and immunohistochemical staining. Growth curve of the C2 gene transfected gastric cancer cells was drawn with the results of MTT assay and cell count. RESULTS: FACS and immunohistochemical staining showed that C2 protein was expressed highly and stably in the C2 gene transfected cells. The growth curve of the C2 gene transfected cells by both MTT and cell count showed that the C2 gene transfected cells had less proliferation than the control cells. The growth cycle of transfected cells was 11 days, while the control was 7 days. CONCLUSION: C2 gene transfection may inhibit the growth of human gastric cancer cells. It might become a potential inhibitor of gastric cancer in the level of protein translation initiation. PMID- 11940310 TI - [The application of Gallyas-Braak stainings in pathologic diagnosis of neurodegenerative diseases]. AB - OBJECTIVE: To evaluate the role of Gallyas silver staining in the diagnosis of neurodegenerative diseases. METHOD: Modified Gallyas-Braak staining method was used to investigate samples of the brain and spinal cord of 22 cases with neurodegenerative disease including Alzheimer's disease (AD), Parkinson's diseas (PD), Pick's disease, diffuse Lewy body disease (DLBD), progressive supranuclear palsy (PSP), diagnosed by clinical and routine pathologic method. 10 cases without clinical symptoms and pathologic abnormalities of the nervous system served as control. RESULT: As compared with Bodian staining, Gallyas-Braak staining demonstrated clearly neurofibrillary tangles in the hippocampus and the cortex of frontal and temperal lobe in all the cases with Alzheimer's disease, 6 cases with dementia of other causes and 3 normal aged. However, global neurofibrillary tangles in the midbrain and the basal ganglia were found only with Gallyas-Braak staining in 4 cases with both dementia and extrapyramidal features. In addition, tuft-shaped astrocytes were shown with this method in the motor cortex, basal ganglia, midbrain of the above 4 cases and astrocytic plaques in the same area in 2 cases of the 4 cases. In this connexion, pathologic findings in 2 of the 4 cases corresponded to PSP and those of the other two cases fufiled the diagnostic criteria of corticobasal degeneration (CBD) Oligodendroglial cytoplasmic inclusions in the white matter of the brain and the spinal cord were founded in 3 of the 4 cases with multiple system atrophy (MSA). This silver staining demonstrated as well a lot of argyrophilic grains in the neuropil of the temporal lobe and the hippocampus in one case with AD. CONCLUSION: Gallyas silver staining could better reveal not only Alzheimer-like neurofibrillary tangles but also different glial inclusions in other neurodegenerative diseases such as PSP, CBD and MSA. Consequently, it is of great value in the pathologic diagnosis and study of such degenerative diseases. PMID- 11940312 TI - [The effects of butyrates and non-steroidal anti-inflammatory agents on HT-29 colonic carcinoma cells]. AB - OBJECTIVE: Adenocarcinoma cells HT-29 were used to investigate whether butyrates and non-steroidal anti-inflammatory agents (NSAIDs, aspirin and NS-398) would be effective adjuvants for inducing apoptosis and to probe its possible mechanisms. METHODS: Prostaglandin E(2) (PGE(2)) isolated from HT-29 cell culture supernants was investigated with ELISA. Flow cytometry was used for estimating apoptosis. The morphology of apoptotic cells was investigated by means of electron microscopy. RESULTS: Butyrates could stimulate the secretion of PGE(2). In contrast, NSAIDs inhibited it to a level below 30 ng/L. Both butyrates and NSAIDs could induce apoptosis. The effects showed the obvious tendency of time- and concentration-dependency (P < 0.05). Combination with two drugs could enhance the above effects. CONCLUSIONS: Butyrates and NSAIDs could induce apoptosis individually. Combination with the two drugs could enhance the above effects by down-regulating cyclooxygenase-2 expression. PMID- 11940313 TI - [Cognitive therapy for patients with refractory irritable bowel syndrome]. AB - OBJECTIVE: To investigate the procedure and tactics used in the cognitive therapy for patients with irritable bowel syndrome (IBS), and to evaluate the efficacy of cognitive therapy in the treatment of refractory IBS. METHODS: A self-control study on the cognitive therapy for 22 patients with refractory IBS symptoms (according to Rome II criteria) was performed. The procedure of cognitive therapy included five steps, namely health education, patients' questioning, relaxing training, dissensitization training, and patients' homework for enforcing the effect of former four steps. The effects of cognitive therapy for IBS were evaluated by improvement of symptom-related-anxiety, index of symptom, quality of life specific for IBS and coping. RESULTS: All 22 cases completed cognitive therapy and first follow-up unit (FFU), at the end of FFU, clinical symptoms in all patients improved (P < 0.05), of them, 81.8% improved significantly (P < 0.001); at 12-months-follow-up, complete remission of clinical symptoms occurred in 72.7% (8/11) patients. Comparison of the scores of symptom-related-anxiety, index of symptom, quality of life specific IBS and coping at the end of 1st follow-up unit with that at basal level, the scores of symptom-anxiety, indexes of severity and frequency of symptoms decreased significantly (P < 0.001, respectively); the scores of depression and anxiety in SCL-90 also decreased significantly (P < 0.001). The scores of active coping rose significantly (P = 0.000). IBS-QOL improved significantly (P < 0.05), of them, dysphoria, body image, food avoidence improved very significantly (P < 0.001, respectively). CONCLUSIONS: Cognitive therapy for patients with refractory IBS is rational and effective. During cognitive therapy, we should follow the therapeutic procedure and the principle of individuation. PMID- 11940314 TI - [The value of gamma-glutamyl transpeptidase mRNA typing in monitoring carcinogensis of heptocytes]. AB - OBJECTIVE: To explore the relationship between the alteration in gamma-glutamyl transpeptidase (GGT) mRNA expression and the development of hepatocellular carcinoma (HCC). METHODS: Three GGT mRNA types (A, B, C) in normal liver tissue, diseased liver tissue without HCC, cancerous and noncancerous tissues from livers with HCC, noncancerous tissues from livers with metastatic tumor and peripheral blood of all the cases were tested by using RT-PCR. RESULTS: In normal livers, the main type of GGT mRNA was type A. In cases with liver diseases but not HCC, the proportion of the types of GGT mRNA was nearly the same as that in normal livers. The proportion of type B was significantly higher in both the cancerous and noncancerous tissues of livers with HCC than in livers without HCC (P < 0.05). The expression of type A in cancerous tissue was significantly lower than that in the tissue of livers without HCC (P < 0.05). Among the 26 cases with HCC, GGT mRNA-B was found in the peripheral blood of 12. In 10 HCC cases with negative AFP, GGT mRNA-B was found in the peripheral blood of 5. CONCLUSIONS: GGT mRNA expression in the human liver might shift from type A to type B during the development of HCC. Fragment analysis of GGT genes might be a sensitive assay to monitor the cancerous change of hepatic cell. PMID- 11940315 TI - [Clinical characteristics of primary biliary cirrhosis: a report of 45 cases]. AB - OBJECTIVE: To study the clinical features of primary biliary cirrhosis (PBC) in order to facilitate cognition of the disease. METHODS: General status, clinical manifestations and laboratory findings of 45 patients with PBC were reviewed. RESULTS: Among the 45 patients, 42 were females and the mean age at diagnosis was (50.8 +/- 8.1) years. The mean time interval between the first visit to physicians to the time of correct diagnosis was about 2 years. The most frequent symptoms were fatigue (66.7%, 30/45), then jaundice (55.6%, 25/45) and pruritus (40%, 18/45). Nine patients (20%) were asymptomatic. Nine patients (20%) were associated with other auto-immune diseases (Sjogren's syndrome and/or rheumatoid arthritis). Serum alkaline phosphatase (ALP) and glutamyl transpeptidase (gamma GT) levels were markedly elevated in all the patients [(498.5 +/- 369.8) IU/L and (595.2 +/- 518.4) IU/L, respectively], whereas ALT and AST levels were mildly to moderately elevated [(83.9 +/- 58.8) IU/L and (100.8 +/- 48.8) IU/L, respectively]. 25 patients (55.6%) had a total bilirubin level >/= 34.2 micromol/L. 40 patients (88.9%) had elevated serum IgM and 43 patients (95.6%) were anti-mitochondrial antibody (AMA)/AMA-M2 positive. CONCLUSION: In China, PBC is probably not so rare as it was thought before. It is mostly found in middle aged women. The most frequent symptom is fatigue and some patients are asymptomatic at early stage. Elevated serum ALP and gamma-GT levels together with positive AMA/AMA-M2 can help to diagnose PBC. Liver biopsy is useful to confirm the diagnosis and differentiate histopathological stages. PMID- 11940316 TI - [A community-based prospective control study of comprehensive intervention in hypertensive patients]. AB - OBJECTIVE: To explore the effect of comprehensive intervention and the prognosis of hypertensive patients in a community. METHODS: To monitor and follow up the hypertencive patients in a community and their families, to intervene comprehensively and prospectively and to make comparison with a control group. RESULTS: The blood pressure of the study group declined from (157.7 +/- 9.0) mm Hg (SBP), (95.0 +/- 16.5) mm Hg (DBP) before entry to (130.6 +/- 11.0) mm Hg (SBP), (80.0 +/- 12.0) mm Hg (DBP), at the end of the study, with significant difference as compared with that of a control group (P < 0.001). The decrement was significant (P < 0.005). Patients' life style was changed and life quality improved. The rate of complications, disability and mortality declined in the study group five times than those in the control group. The total score of SCL-90 was significantly declined in the study group than control group. CONCLUSIONS: The beneficial effect of community and family intervention and health knowledge education was confirmed in this study. There is still a long way to go for popularizing public health education, training professional health personnel and enhancing the compliance of hypertensive patients to treatment. PMID- 11940317 TI - [The application of transcranial Doppler in detecting diabetic cerebral macroangiopathy and microangiopathy]. AB - OBJECTIVE: To evaluate the application of transcranial Doppler (TCD) in the detection of cerebral hemodynamic changes in diabetic macroangiopathy and microangiopathy. METHODS: 81 of diabetic patients and 62 age-, gender- matched healthy controls were monitored with TCD in this study. The pulsatility index (PI) and mean flow velocities (Vm) between the patients and healthy controls, between the patients with microangiopathic complications and patients without complication, were analyzed respectively. RESULTS: The PIs of arteries and Vms of the extracranial internal carotid artery (ICA), middle cerebral artery (MCA) and basilar artery (BA) were significantly higher in diabetic patients than those in healthy controls (P < 0.001), while PIs of ICA and MCA but not BA were significantly increased in patients with complications than those in noncomplicated patients (P < 0.001). CONCLUSIONS: It indicated that PI and Vm might be two useful markers for the detection of diabetic cerebrovascular changes. Marked increase of PIs of ICA and MCA might reflect the microangiopathic changes of cerebral vessels in diabetic patients. PMID- 11940318 TI - [Inhibition of telomerase activity and induction of apoptosis in lung cancer cell by human telomerase reverse transcriptase gene antisense oligodeoxynucleotide]. AB - OBJECTIVE: To evaluate the effect of human telomerase reverse transcriptase (hTERT) gene antisense oligodeoxynucleotide (ASODN) on telomerase activity and apoptosis in lung cancer cell A549 line. METHOD: Cultured cells were divided into three groups: ASODN (5'-GCGCGGCAGCGCGGGGGTGGCCG-3'), sense oligodeoxynucleotide (5'-CGGCCACCCCCGCGATGCCGCGC-3', SODN) and control. The concentration of oligodeoxynucleotide and lipsome was 10 micromol/L and 16 mg/L respectively. The changes of telomerase activity, expression of hTERT mRNA and protein and apoptosis were observed in different groups. RESULTS: hTERT mRNA and protein levels, as well as telomerase activity began to be down-regulated or inhibited when A549 cells were treated with ASODN for 72 h, but apoptosis was induced until cells were treated for 21 days with ASODN (rate of apoptosis was 0.88%, 2.77% and 12.00% for the control, SODN and ASODN group respectively). CONCLUSION: It is suggested that hTERT ASODN might specially inhibit telomerase activity at translation level in lung cancer cells and it is further proved that the hTERT gene have significant correlation with telomerase activity. Further evidence is needed to prove whether hTERT ASODN might be a potential tool for the treatment of lung cancer. PMID- 11940320 TI - [The relationship between expression of lung resistance-related protein gene or multidrug resistance-associated protein gene and prognosis in newly diagnosed acute leukemia]. AB - OBJECTIVE: To evaluate the relationship between the expression of lung resistance related protein (lrp) gene or multidrug resistance-associated protein (mrp) gene and prognosis in untreated acute leukemia (AL) patients. METHODS: Reverse transcriptase-polymerase chain reaction (RT-PCR) was used to examine the expression of lrp and mrp gene in 58 newly diagnosed AL patients. RESULTS: The positive rate of lrp and mrp gene expression in newly diagnosed acute lymphocytic leukemia (ALL) group was 15.0% and 40.0% and it was 15.8% and 42.1% in newly diagnosed acute nonlymphocytic leukemia (ANLL) group. In both the ALL and ANLL groups, the difference of the first complete remission rate in lrp negative and in lrp positive patients was not significant (P > 0.05), the same results were found with mrp gene. The difference of the first complete remission rate between lrp(+)/mrp(+) and lrp(-)/mrp(-) patients was significant (P < 0.05). There was no relationship between lrp and mrp gene. CONCLUSION: Neither lrp nor mrp gene as a single indicator to forecast original multidrug resistance is sensitive, but lrp combined with mrp as one indicator will be sensitive. PMID- 11940319 TI - [The effects of paraoxonase-1 and oxidized low density lipoprotein on nephropathy in type-2 diabetes mellitus]. AB - OBJECTIVE: To explore the effect of paraoxonase-1 (PON-1) and oxidized low density lipoprotein (ox-LDL) in diabetic nephropathy (DN). METHODS: Serum PON-1 activity was measured by using phenyl acetate as a substrate and the concentration of plasma ox-LDL was determined by ELISA in 91 cases of type 2 diabetes mellitus (DM). Simultaneously, serum NO, VWF and GMP 140 were also determined. The patients of DM were divided into three groups according to urine albumine excretion rate (UAER) (DM I: UAER < 30 mg/24 h; DM II: UAER 30 approximately 300 mg/24 h; DM III: UAER > 300 mg/24 h) and matched with normal controls (NC). RESULTS: In the three groups of DM, PON-1 were markedly decreased and ox-LDL were obviously increased (P < 0.01). Both of their levels were different in the three groups of DM (P < 0.05). PON-1 were negatively correlated with ox-LDL (r = -0.467, P < 0.01). There were negative correlation between PON-1 and urine albumin (r = -0.697, P < 0.01), but ox-LDL was positively correlated with urine albumin (r = 0.747, P < 0.01). Serum nitrites (NO) level in DM I was significantly higher than that in NC, but the NO level in DM II and DM III was lower than that in DM I and NC. GMP 140 and VWF levels in all the 3 DM groups were higher than that in NC. PON-1 was positively correlated with NO (r(s) = 0.326, P < 0.01), and negatively correlated with VWF (r(s) = -0.365, P < 0.01). ox-LDL was negatively correlated with NO (r(s) = -0.196, P < 0.05), but positively correlated with VWF and GMP 140 (r(s) = 0.294, P < 0.05; r(s) = 0.669, P < 0.01 respectively). Logistic regression showed that ox-LDL was a dangerous factor for DN. CONCLUSION: PON-1, ox-LDL, NO, endothelial and platelet function are connected with DN; they affect one another and influence the occurrence and development of DN together. PMID- 11940321 TI - [Antidotal effects of vitamin B(6) and sodium dimercaptopropane sulfonate on acute poisoning with tetramethylene disulphotetramine in animals]. AB - OBJECTIVE: The study was designed to examine the antidotal effects of vitamin B(6) (Vit B(6)) with sodium dimercaptopropane sulfonate (Na-DMPS) on acute poisoning with tetramethylene disulphotetramine (TETS) in animals. METHODS: Through the acute toxicity experiments, the antidotal effects of Vit B(6) with Na DMPS on mice immediately and 10 minutes after poisoning were observed. RESULTS: Used of Vit B(6) with Na-DMPS in early stage showed excellent inhibition of tetanic convulsion in acute poisoning with TETS in mice and can reduce very significantly the death rate (P < 0.001). Vit B(6) with Na-DMPS used 10 minutes after poisoning also significantly elongate the death time (68.8 +/- 15.2) and reduce significantly the death rate (60%) then control group's death time (31.6 +/- 7.8) and the death rate (100%) on acute poisoning in mice (P < 0.01). CONCLUSION: Antidotal effects of Vit B(6) with Na-DMPS were significant on acute TETS poisoning in animals. Vit B(6) with Na-DMPS is an excellent antidote for acute TETS poisoning. It is suggested that Vit B(6) with Na-DMPS may be clinically used to rescue patients poisoned by TETS. PMID- 11940322 TI - Primary intracoronary stenting in comparison with intravenous rt-PA thrombolysis plus rescue intracoronary intervention in patients with acute myocardial infarction. AB - OBJECTIVES: To compare primary stenting in the infarct-related coronary artery with intravenous rt-PA therapy plus rescue intracoronary stenting. METHODS: Ninety-eight patients with a first acute myocardial infarction (AMI) were randomly treated with primary intracoronary stenting (primary stenting group) or with intravenous rt-PA therapy plus rescue intracoronary stenting (thrombolysis plus stenting group). Thrombolysis in myocardial infarction (TIMI) flow grade was assessed by angiography in emergency, and cardiac function (left ventricular ejection fraction, LVEF) was calculated by echocardiography before discharge between the two groups. RESULTS: There were 47 patients (97.91%) in primary stenting group and 50 patients (100%) in thrombolysis plus stenting group had achieved TIMI grade 2 - 3 flow after the procedure. But the former had more cases (93.8%) of TIMI 3 flow than that of latter (60.0%, P = 0.0001). There was no difference between the two groups in cardiac events during hospitalization. But the patients in primary stenting group had better cardiac function (LVEF 0.62 +/- 0.14 vs. 0.50 +/- 0.12, respectively, P = 0.0001) between the two groups. CONCLUSIONS: Primary intracoronary stenting may improve myocardial reperfusion in emergency and inhibit the decline of cardiac function after AMI in comparison with intravenous rt-PA thrombolysis plus rescue intracoronary stenting. PMID- 11940323 TI - Cutting balloon angioplasty for treatment of coronary in-stent restenosis: immediate results and 6-month outcomes. AB - OBJECTIVE: To determine the mid-term effects of cutting balloon angioplasty (CBA) on in-stent restenosis. METHODS: A total of 69 patients with in-stent restenosis were divided into 2 groups randomly: cutting balloon angioplasty and plain old balloon angioplasty. The mechanisms of restenosis and dilation results were determined by quantitative coronary angiography and intravascular ultrasound. Follow-up was performed. RESULTS: The procedural success rate was 100% without death and acute closure. One patient experienced dissection at the distal end of the stent and needed another stent. The mean follow-up period was 6.7 +/- 2.3 months. The final re-restenosis rate was 15% and 18% at 3 months and 6 months respectively, markedly lower than after plain old balloon angioplasty (38% and 43%). Acute gain by intravascular ultrasound (IVUS) was 1.72 +/- 0.52 mm after cutting balloon angioplasty, higher than 1.15 +/- 0.54 mm after plain old balloon angioplasty. The lumen diameter late loss in the cutting balloon group was 0.26 +/- 0.05 mm and 0.38 +/- 0.06 mm at 3 months and 6 months respectively, significantly lower than for those in conventional balloon group (0.78 +/- 0.19 mm and 0.89 +/- 0.16 mm, respectively, P < 0.001). As shown by IVUS, the main mechanism of cutting balloon angioplasty was marked reduction of plaque area without significant increase of vessel area (less vessel trauma). CONCLUSION: Cutting balloon angioplasty is feasible and effective for the treatment of in stent restenosis with less vessel trauma. PMID- 11940324 TI - Immunization with beta(1)-adrenoreceptor peptide induces cardiomyopathy-like changes in rabbit hearts. AB - OBJECTIVE: To investigate the importance of autoimmunity against beta(1) adrenoreceptor in the pathogenesis of dilated cardiomyopathy (DCM). METHODS: Fourteen rabbits were divided equally into two groups. Rabbits in the immunized group (n = 7) were immunized monthly for one year with synthetic peptide corresponding to the second extracellular loop of the beta(1)-adrenoreceptor and adjuvant. Control rabbits received the mixture with the same procedure as described except with a substitution of saline for the corresponding peptide. During the study period, all rabbits were bled to assay the titers of antipeptide antibody and left ventricular ejection fractions (LVEFs) were measured by emission computed tomography. At the end of experiment, invasive cardiac function was measured and morphologic examinations were done. RESULTS: High titers of antipeptide antibody were found in the sera from immunized rabbits throughout the study period in contrast to those from control rabbits. LVEFs were significantly higher in immunized rabbits than those of the control group at the 4th and 6th month. At the end of the experiment, the maximal rates of rise and decline of ventricular pressure of the immunized group were significantly lower than those of the control group. Morphological changes were found in immunized rabbits such as the enlargement of ventricles, myofibrillar lysis and necrosis, mitochondria swelling and condensation. No obvious alterations were noted in hearts of control rabbits. CONCLUSION: Autoimmunity against the beta(1)-adrenoreceptor may be involved in the pathogenesis of dilated cardiomyopathy and beta(1)-adrenoreceptor antibody may play a role in the process. PMID- 11940325 TI - CD59 prevents human complement-mediated injuries in isolated guinea pig hearts. AB - OBJECTIVE: To assess complement-mediated myocardial injury on isolated guinea pig working hearts and cardioprotective effects of CD59. METHODS: Using a modified Langendorff apparatus, isolated guinea-pig working hearts were perfused with a modified Krebs Henseleit buffer containing 3% heat-inactivated human plasma and zymosan (IPZ) (control) (n = 10), 3% normal human plasma and zymosan (NPZ) (n = 10), or 3% normal human plasma and zymosan and 1.5 microg/ml CD59 (NPZC) (n = 10), respectively. Epicardial electrocardiogram (ECG), cardiac output (CO), coronary arterial flow (CF), maximum left ventricular developed pressure (LVP(max)), maximum left ventricular developed pressure increase rate (+ dp/dt(max)), maximum left ventricular developed pressure decrease rate (- dp/dt(max)) and heart rate (HR) were recorded at 0, 15, 30, 45 and 60 min of treatment. After the experiment, immunohistochemical examination was performed to detect the presence of C3a or C5b-9 in the myocardium of the isolated hearts. RESULTS: Compared the IPZ group, hearts treated with NPZ showed a slight depression on ST segments of epicardial ECG at 15 min, a significant elevation between 30 min to 60 min, a decrease in CF, CO, LVP(max), + dp/dt(max) and - dp/dt(max), and an increase in HR at 15 min. The observed alterations in CF, CO, LVP(max), + dp/dt(max) and - dp/dt(max) remained decreased, while the HR remained increased until the end of the protocol. The all above parameters of hearts treated with NPZC were similar to the control group (IPZ) at any given time. Immunohistochemical examination showed positive signals of C3a and C5b-9 in the myocardium of hearts treated with NPZ. C3a was positive in NPZC, and C3a and C5b 9 were negative in IPZ. CONCLUSIONS: Activated human complements directly damage isolated guinea pig working hearts, and CD59 offers a significant protection against the injuries. PMID- 11940326 TI - Proportional assist ventilation: methodology and therapeutics on COPD patients compared with pressure support ventilation. AB - OBJECTIVE: To investigate the impact of proportional assist ventilation (PAV) on tolerance and breathlessness in ventilated chronic obstructive pulmonary disease (COPD), and to describe the patient-ventilator interaction, hemodynamic state, breathing pattern and work of breath during PAV and pressure support ventilation (PSV). METHODS: Ten intubated COPD patients on weaning from mechanical ventilation were studied. Elastance and resistance were measured by both the inspiratory-hold technique during a brief period of volume control ventilation and runaway technique during PAV. Each assistance level of PAV (80%, 60% and 40%) and PSV was selected randomly. Patients' response, hemodynamics, blood gas and lung mechanics were monitored. RESULTS: Tidal volume and respiratory rate didn't change in a consistent manner as the level of assist was decreased (P > 0.05). With the level of assist increasing, peak inspiratory pressure was increasing significantly (P < 0.05), while patients' work of breath had the tendency to decrease (P < 0.05). A significant difference in the Borg Category Scale was observed between PAV and PSV (0.50 [1.50] vs. 0.75 [2.00], P < 0.05) at the same degree of respiratory muscle unloading. PaCO(2) was significantly higher on PAV (54 [23] mm Hg) than on PSV (48 [23] mm Hg) (P < 0.05). Peak inspiratory pressure on PAV was significantly lower than on PSV (16 +/- 4 cm H(2)O vs. 21 +/- 3 cm H(2)O, respectively, P < 0.05). Hemodynamics and oxygenation remained unchanged. CONCLUSIONS: PAV is a feasible method for supporting ventilator-dependent patients and was well tolerated. It can improve the breathing pattern and reduce inspiratory effort. At the same degree of respiratory muscle unloading, PAV can be implemented at much lower peak inspiratory pressure than PSV. It can also apply proportional pressure support according to the patients' ventilatory demand. PMID- 11940327 TI - Assessment of polymerase chain reaction and serology for detection of chlamydia pneumoniae in patients with acute respiratory tract infection. AB - OBJECTIVE: To study Chlamydia pneumoniae (C. pneumoniae) infection in 110 patients with respiratory tract infection admitted to our hospital from January to December 1995 in Nanjing. METHODS: Sputum and throat swab specimens were taken and C. pneumoniae DNA was detected by using polymerase chain reaction (PCR) with the HM-1-HR-1 primer pair. At the same time, serum samples were taken and immunoglobulin G and M (IgG and IgM) fractions of antibodies to C. pneumoniae were studied by microimmunofluorescence test. RESULTS: Prevalence of specific IgG was 70% in patients with respiratory tract infection. Seventeen patients (15.5%) were serologically diagnosed as having recent C. pneumoniae infections and 12 patients (10.9%) had positive PCR in sputum and/or swab specimens. The total positive rate was 22.7% (25/110) detected by PCR combined with serological tests. Acute infection of C. pneumoniae was common in patients with asthma (57.1%), pneumonia (35.0%), COPD (25.9%) and bronchitis (25.0%). Clinical features between C. pneumoniae infection and non-C. pneumonia infection showed no significant differences. CONCLUSIONS: Chlamydia pneumoniae is an important pathogen that causes infection of the human respiratory tract and attention should be drawn to this special illness. PMID- 11940328 TI - Distribution of ventilation and hemodynamic effects of different ventilatory patterns. AB - OBJECTIVE: To compare the influence of cardiac-pulmonary function on clinical acute respiratory failure patients using Proportional assist ventilation (PAV), Pressure support ventilation (PSV) and intermittent positive pressure ventilation (IPPV). Here, we also describe some our experience with the clinical use of PAV. METHODS: Using the IPPV mode in ten acute respiratory failure patients, calculate Elastance (Ers) and Resistance (Rrs), then change to PSV, set inspiratory positive airway pressure (IPAP) according to IPPV, so that tidal volume (V(T)) is the same as that of IPPV. We then changed the mode into PAV and set the assist ratio according to PSV, so that V(T) and Ppeak were the same as that of PSV. Then we observed the changes of respiratory mechanics, blood gas levels and hemodynamics during ventilation. RESULTS: Compared with PSV and IPPV, peak pressure (Ppeak) of PAV was markedly lower while V(T) was similar; work of breathing of patient (WOBp), and work of breathing of ventilation (WOBv) were also lower; center vein pressure (CVP) and pulmonary capillary wedge pressure (PCWP) of PAV were markedly lower than that of IPPV while V(T) were similar. Compared with PSV, V(T), mean blood pressure (mBP) and cardiac output (CO) of PAV were higher. Mean pulmonary artery pressure (mPAP) and WOBp of PAV were lower while Ppeak was similar; the differences in WOBp were notable. CONCLUSIONS: For clinical acute respiratory failure patients, compared with PSV and IPPV, PAV has lower airway pressure, less WOBp and less influence on hemodynamics. PMID- 11940329 TI - Genetic variation of mannose-binding protein associated with glomerular immune deposition in IgA nephropathy. AB - OBJECTIVE: To investigate the relationship between codon 54 gene polymorphism of the host defense molecule, mannose-binding protein (MBP), and the patterns of glomerular immune deposition in IgA nephropathy (IgAN). METHODS: IgAN patients with different patterns of glomerular immune deposition were selected and divided into two groups. Group A consisted of 77 patients with glomerular IgA and C3 deposits, and Group AGM consisted of 70 patients with glomerular IgA, IgG, IgM, C3 and Clq deposits. Clinical features and laboratory relevant data of all patients were collected. One-hundred and forty healthy adults were recruited as normal controls. The MBP gene codon 54 GGC/GAC polymorphism was investigated by using polymerase chain reaction and restriction fragment length polymorphism. RESULTS: The genotype frequency of GGC/GAC heterozygotes was significantly higher in Group AGM as compared with that of Group A (41.4% vs 19.5%, P < 0.01) or normal subjects (41.4% vs. 26.4%, P < 0.05), while no difference was found in the distribution of MBP genotypes between Group A and normal subjects. GAC allele frequency was also higher in Group AGM than that in Group A (0.24 vs. 0.14, P < 0.05) or normal subjects (0.24 vs. 0.15, P < 0.05). The variant allele (GAC) was markedly associated with Group AGM (OR = 1.95, 95% CI: 1.06 - 3.58). In both Group A and Group AGM, more patients carrying the variant allele had episodes of upper respiratory or gastrointestinal infections prior to the onset of IgAN than those with wild homozygotes (GGC/GGC). CONCLUSIONS: Genetic variation of the host defense molecule, MBP, may be involved in the formation of the diverse patterns of glomerular immune deposition in IgAN. The variant allele of the MBP gene may partially account for abundant immune deposits in some IgAN patients. PMID- 11940330 TI - The effect of L-arginine on the progression of chronic renal scarring in remnant kidney. AB - OBJECTIVE: To investigate the effect of L-arginine (L-arg) on early compensatory renal growth (CRG), tubulointerstitial accumulation of extracellular matrix (ECM), long term survival rate and renal scarring in rats with 5/6 nephrectomy (SNx). METHODS: The experiment included four groups of rats (n = 5 each group): (1) Sham group, (2) SNx group, (3) SNx + L-arg group, and (4) Sham + L-arg group (L-arg 1% in drinking water). Parameters related with CRG and early tubulointerstitial expression of ECM and alpha smooth muscle actin (alphaSMA) were evaluated by immunohistochemistry at day 30. The survival rate and the extent of renal scarring in the rats were observed at day 120. RESULTS: L-arg significantly increased the early CRG of SNx rats as determined by the wet kidney weight (P < 0.05), total protein (P < 0.01), and DNA content (P < 0.001) of the remnant kidney. The cell proliferation in the tubulointerstitium as determined by immunostaining for proliferative cell nuclear antigen (PCNA) also markedly increased (P < 0.05). Furthermore, glomerular volume (Vg) in the SNx rats treated by L-arg increased 16% compared with that in the SNx group (P = 0.06). More specifically, L-arg significantly increased the interstitial immunostaining for collagen III, IV, and fibronectin (P < 0.05, 0.01, respectively), which were positively correlated with the increased expression of alpha-SMA (P < 0.001, respectively). In the long term experiment, L-arg significantly reduced the survival rate (P < 0.05) and increased the severity of renal scarring in the SNx rats (P < 0.05). CONCLUSION: This preliminary study suggested that early supplementation with L-arg might exacerbate the progression of renal scarring in rat remnant kidney. PMID- 11940331 TI - Recombinant human Flt3 ligand exerts both direct and indirect effects on hematopoiesis. AB - OBJECTIVE: To investigate the direct effects of the Flt3 ligand (FL) on hematopoiesis, such as the stimulation of the formation of hematopoietic colonies and the proliferation of dendritic cells, as well as the indirect stimulation of hematopoiesis, especially via the proliferation of endothelial cells. METHODS: Mononuclear cells from human cord blood were plated in methylcellulose medium containing different cytokines to induce hematopoietic colony formation. Dendritic cells (DCs) were induced from the mononuclear cells with a cytokine cocktail with or without recombinant human soluble FL (rhFL; 100 ng/ml). The Flt3 receptors on the surface of a human microvascular endothelial cell line (ECV) were analyzed by flow cytometry. The proliferation of ECV stimulated by rhFL was measured with the microculture tetrazolium assay. The levels of FL, IL-6, IL-8, G CSF and GM-CSF in the supernatant of ECV cultures were measured by enzyme linked immunoabsorbent assay (ELISA). RESULTS: rhFL stimulates colony formation from cord blood when used as a sole stimulant. FL in combination with other cytokines increased colony formation significantly. The number of DCs was approximately 2.5 times higher when rhFL was used. rhFL stimulates the proliferation of ECV on which Flt3 receptors are expressed. Furthermore, ECV secretes FL, IL-6, IL-8, G CSF and GM-CSF, which were augmented by tumor necrosis factor-alpha and rhFL. CONCLUSIONS: rhFL enhances hematopoietic colony formation and DC proliferation from human cord blood cells. FL not only stimulates the proliferation of ECV, but is also secreted by ECV. FL may exert direct and indirect effects on hematopoiesis. PMID- 11940332 TI - Phenotypic analysis of affected peripheral erythroid for CD59 in paroxysmal nocturnal hemoglobinuria. AB - OBJECTIVE: To determine whether affected reticulocytes could be a reliable marker for the diagnosis of paroxysmal nocturnal hemoglobinuria (PNH), we analyzed CD59 antigen expression on the membranes of reticulocytes and erythrocytes. METHODS: We studied 10 PNH patients and 5 healthy volunteers by two-color flow cytometry with a membrane permeable fluorescent dye, thiazole orange (TO), and anti-CD59 monoclonal antibodies (MoAb). TO was introduced to gate reticulocytes and anti CD59 MoAb were used to identify glycosylphosphatidylinositol (GPI)-deficient cells. RESULTS: Cells from healthy individuals were only CD59 positive. However, in all PNH patients, CD59-antigen expression could be divided into 3 types: type I cells (CD59 normally positive), type II cells (CD59 partly positive) and type III cells (CD59 negative). The majority of reticulocytes belonged to type III cells, GPI-deficient cells (61.0%). In addition, the percentage of affected reticulocytes was higher than that of erythrocytes. CONCLUSIONS: Analyzing PNH reticulocytes was important, because most patients had elevated numbers of reticulocytes, which represent more closely the recent erythroid output of BM. However, circulating mature erythrocytes were subject to complement-mediated intravascular lysis. Therefore, the percentage of abnormal erythrocytes may not accurately reflect the proliferation rate of normal and abnormal erythroid progenitor cells. Thus, affected reticulocytes could be a more reliable indicator for the diagnosis of PNH than mature erythrocytes. PMID- 11940333 TI - Polymorphisms and functions of the aldose reductase gene 5' regulatory region in Chinese patients with type 2 diabetes mellitus. AB - OBJECTIVE: To screen the 5' regulatory region of the aldose reductase (AR) gene for genetic variabilities causing changes in protein expression and affecting the promoter function. METHODS: The screenings were carried out by polymerase chain reaction-single strand conformation polymorphism (PCR-SSCP). All SSCP variants were submitted for DNA sequencing and inserted into the plasmid chloromycetin acetyl transferase (CAT) enhancer vector. The constructs were used to transfect Hela cells, and CAT assays were performed to assess promoter activity. Gel mobility shift and footprinting assays were also performed to determine the interaction between the DNA and nuclear proteins. RESULTS: Two polymorphisms, C ( 106) T and C (-12) G, were identified in the regulatory region in 123 Chinese control subjects and 145 patients with type 2 diabetes mellitus. The frequencies of genotypes WT/WT, WT/C (-12) G and WT/C (-106) T were not significantly different between the subjects and patients. In the patients with and without retinopathy, frequencies of WT/C (-106) T were 31.5% and 17.5% (P < 0.05) respectively, and the frequencies of WT/C (-12) G were 10.5% and 2.5% (P > 0.05) respectively. The total frequency of WT/C (-12) G and WT/C (-106) T in patients with retinopathy was 41.8%, significantly higher than that (20.0%) in patients without retinopathy (P < 0.025). The relative transcription activities of the wild-type, the C (-12) G and the C (-106) T were 15.7%, 31.0% and 32.2%, respectively. The results of DNA-protein interaction assays showed that these variations did not change the binding site of DNA with trans-acting factors. CONCLUSION: The polymorphisms C (-12) G and C (-106) T strongly associated with diabetic retinopathy in the Chinese population have been identified in the regulatory region of the aldose reductase gene. PMID- 11940334 TI - beta-fibrinogen gene -455A/G polymorphism and plasma fibrinogen level in Chinese stroke patients. AB - OBJECTIVES: To investigate the relationship between the beta-fibrinogen gene 455A/G polymorphism and plasma fibrinogen level and to determine the influence of the mutation on ischemic stroke. METHODS: Ninety-one patients (63.5 +/- 10.1 years) with ischemic stroke and 74 elderly control subjects (60.6 +/- 10.8 years) without any thromboembolic events and 98 healthy blood donators as young control (37.5 +/- 13.3 years) were enrolled in this trial. The beta-fibrinogen gene 455A/G polymorphism was analyzed for all subjects by PCR-RFLP with the restrictive enzyme Hae III, while plasma fibrinogen levels were obtained from the prothrombin time (PT) assay. For statistical analysis, the parameters were compared between any two different groups by the unpaired Student's t test and the Chi-square test. Before analysis, log transformations for concentrations of fibrinogen were carried out. RESULTS: H2 allele frequency was higher in male ischemic stroke patients than in the elderly control (22.7% vs 7.1%, chi(2) = 5.56, P < 0.02). There was no significant difference between the female groups. In those patients without any thromboembolic events (both elderly and young control groups), the frequency of H2 decreased with age (< or = 40, 21.3%; 41 - 59, 15.4%; and > or = 60, 10.2%). In the male elderly and young control groups, the level of plasma fibrinogen was lower in the H1H1 genotype (287 +/- 96 mg/dl and 234 +/- 58 mg/dl) than in H1H2 and H2H2 (331 +/- 44 mg/dl and 307 +/- 55 mg/dl; t = 2.53 and 9.67, P < 0.05). In the female elderly groups, this tendency was not found. CONCLUSION: Plasma fibrinogen expression is affected by the beta fibrinogen gene -455A/G polymorphism, and the H2 allele may be a risk factor for ischemic stroke in Chinese males. PMID- 11940335 TI - Screening for tetrahydrobiopterin deficiency among hyperphenylalaninemia patients in Southern China. AB - OBJECTIVES: To assess the incidence of tetrahydrobiopterin (BH4) deficiency among patients with hyperphenylalaninemia (HPA) in southern Chinese and evaluate clinical outcome and gene mutations in tetrahydrobiopterin deficient patients. METHODS: Urinary neopterin (N) and biopterin (B) was analyzed in 87 patients with hyperphenylalaninemia by high-performance liquid chromatography. Further combined loading tests with phenylalanine (Phe) (100 mg/kg) and tetrahydrobiopterin (BH4) (7.5 mg/kg) were performed in suspected patients with abnormal urinary pterin profiles. Gene mutation analysis was performed for patients with BH4 deficiency and their parents. BH4 deficient patients were treated with BH4 and neurotransmitter precursors after diagnosis. Blood phenylalanine levels, clinical symptoms and mental development were followed up. RESULTS: Eleven patients were diagnosed as having BH4 deficiency caused by 6-pyruvoyl tetrahydropterin synthase (PTPS) deficiency. The incidence of tetrahydrobiopterin (BH4) deficiency among patients with hyperphenylalaninemia (HPA) in southern Chinese was 10%. Combined loading tests with phenylalanine and oral BH4 were done in 4 of 11 patients and their phenylalanine levels were decreased to normal 4 - 6h after BH4 administration. Four different mutations (P87S, N52S, D96N and G144R) in the PTPS gene were detected in 5 families. Five PTPS-deficient patients were treated with synthetic BH4, neurotransmitter precursors (L-dopa plus carbidopa, and 5 hydroxytryptophan). They had satisfactory physical and mental development after treatment. One patient with partial PTPS deficiency had normal growth and mental development without treatment. CONCLUSIONS: Our results emphasize that screening for BH4 deficiency should be carried out in all patients with hyperphenylalaninemia in order to minimize the misdiagnosis. Patients with BH4 deficiency should be treated early with BH4 and a combination of neurotransmitter precursors. PMID- 11940336 TI - Human breast carcinoma xenografts in nude mice. AB - OBJECTIVE: To investigate spontaneous metastasis, micrometastasis and genetic stability in human breast carcinoma xenografts in nude mice. METHODS: Intact tissue from surgical specimens from breast carcinoma patients was xenografted into nude mice and transplanted from generation to generation. Cells from the xenografts were cultured in vitro and retransplanted into nude mice. Microsatellite DNA in the genome of human breast carcinomas, xenotransplanted tumors and metastases in nude mice were analyzed at three microsatellite loci. RESULTS: The tumorigenicity of orthotopic xenotransplantation was 88.6% (31/35), with a metastatic rate of 41.9% (13/31). Cells from xenotransplants were successfully cultured in vitro. The taking rate of retransplantation into nude mice and the spontaneous lung metastasis rate were both 100% (10/10). Microsatellite DNA sequences in the genome of xenotransplanted tumors and metastases in nude mice were identical with that of the original human breast carcinoma at three microsatellite loci. CONCLUSIONS: Tumorigenicity and metastatic potential can be improved in human breast carcinoma xenografts using intact fresh tumor tissue and orthotopic grafts. Xenotransplanted tumors and tumors after serial passage maintained the genetic stability. The detection of microsatellite DNA may identify micrometastases in a nude mouse model. PMID- 11940337 TI - In situ transduction of cytosine deaminase gene followed by systemic use of 5 fluorocytosine inhibits tumor growth and metastasis in orthotopic prostate cancer mouse models. AB - OBJECTIVE: To investigate the antitumor and anti-metastatic effect of in situ transduction of adenovirus encoding cytosine deaminase (AdCD) followed by the systemic use of 5-fluorocytosine (5-FC) in the orthotopic (o.t.) prostate cancer mouse model. METHODS: The o.t. prostate cancer model of C57BL/6 mouse was developed by o.t. inoculation of RM-1 cells to the subcapsular area of the prostate gland. In situ transduction of the CD gene, followed by systemic use of 5-FC at a daily dosage of 300 mg/kg for 14 days, was performed two days later. RESULTS: Compared with mice treated with Adbeta-gal/5-FC, 5-FC and PBS, mice of the o.t. model receiving in situ treatment of AdCD/5-FC had significant prolongation of survival and suppression of local tumor growth. More importantly, pathological observations showed that metastatic activity occurred in all mice of the PBS, 5-FC and Adbeta-gal groups including metastasis to the iliac lymph node (10/10, 10/10, 10/10) and the lung (8/10, 7/10, 7/10). However, only two out of ten had iliac lymphatic metastasis in the AdCD/5-FC group with no systemic or preaotic lymphatic metastasis, suggesting a strong metastatic inhibitory effect. CONCLUSIONS: In situ transduction of AdCD followed by systemic use of 5-FC leads to the inhibitory effect on tumor growth and metastatic activity in the o.t. mouse model of prostate cancer. Clinically, it may be possible to treat metastatic or recurrent prostate cancer with a novel gene therapy using in situ injection techniques in future. PMID- 11940338 TI - Predictors of atrial fibrillation after coronary artery bypass graft surgery. AB - OBJECTIVE: To identify the clinical predictors of atrial fibrillation (AF) after coronary artery bypass grafting (CABG). METHODS: 322 consecutive patients who had undergone isolated CABG were reviewed. Preoperative, intraoperative and postoperative data were collected. Patients were grouped according to whether AF appeared postoperatively. RESULTS: AF occurred in 75 patients (23.3%). Most cases of AF (85.6%) appeared on or before the third postoperative day. The mean age for patients with AF was 62.5 years compared with 56.7 years for patients without AF (P < 0.05). The mean aortic crossclamp time for patients with AF was 67 min compared with 60.3 min for patients without AF (P < 0.05). The mean duration of cardiopulmonary bypass for patients with AF was 109.6 min compared with 97.3 min for patients without AF (P < 0.05). The mean duration of mechanical ventilation for patients with AF was 19.1 h compared with 15.7 h for patients without AF (P < 0.05). Multivariate logistic regression analysis was used to identify the following independent predictors of postoperative AF (P < 0.05): age > or = 65 years (OR 2.7; 95% CI 1.5 to 5.1), lesions in the right coronary artery (OR 2.5; 95% CI 1.4 to 4.5), and early postoperative withdrawal of beta blocker (OR 3.9; 95% CI 2.1 to 7.7). CONCLUSIONS: AF remains the most common complication after CABG. Age and lesions in the right coronary artery can influence the incidence of AF, and beta blocker and magnesium may be the most economical and effective prevention for AF early after CABG. PMID- 11940339 TI - Long-term prognostic analysis of thymectomized patients with myasthenia gravis. AB - OBJECTIVE: To study the factors affecting the long-term prognosis of patients with myasthenia gravis (MG) after thymectomy. METHODS: 170 MG patients who had undergone thymectomies were studied retrospectively. Among them, 124 patients received long-term follow-up for more than 40 months postoperatively. The COX regression analysis model was used to analyze the factors that may influence the long-term prognosis. These factors included thymus pathology, patient gender, age, duration of disease at the time of surgery, preoperative Osserman classification and medication. RESULTS: The research showed that thymus pathology was the single independent factor that affected the postoperative long-term prognosis. The long-term survival rates differed significantly with thymus pathological types: hyperplasia > benign thymoma > atrophy > malignant thymoma (P < 0.05). CONCLUSION: The different pathological types of the thymus were the important factor affecting long-term survival in MG patients after thymectomy. PMID- 11940340 TI - Establishment of multidrug-resistance cell line C(6)/adr and reversal of drug resistance. AB - OBJECTIVE: To investigate the mechanism of multidrug resistance (MDR) in a human glioma cell and methods for overcoming multi-drug resistance. METHODS: MDR cell line C(6)/adr was established. The expression of the mdr-1 gene and its P glycoprotein (P-gp) in the C(6)/adr cell line was observed by RT-PCR and flow cytometry. The reversal of MDR by verapamil, erythromycin, dihydropyridine, P-gp monoclonal antibody and Salvia miltiorrhiza (SM) was studied by microtiter tetrazolium (MTT) assay or by high performance liquid chromatographic assay. RESULTS: The mdr-1 gene of the C(6)/adr cell line was positive, over-expressing P gp. The drug-resistance of the C(6)/adr cell lines could be partly reversed by 2 6 microg/ml of verapamil, 50 - 100 microg/ml of erythromycin, or 5 microg/ml of dihydropyridine. As concentration increased, they had a better effect. Among these drugs, 100 microg/ml of erythromycin had the best result of reversal. Dihydropyridine 1 microg/ml, P-gp monoclonal antibody and SM had no effect. CONCLUSION: The mdr-1 gene and its expression might be associated with the MDR of glioma cells. Verapamil, erythromycin and dihydropyridine could reverse the MDR of glioma cells. PMID- 11940341 TI - Complication of cesarean section: pregnancy on the cicatrix of a previous cesarean section. AB - OBJECTIVE: To probe into the clinical manifestation, diagnosis, as well as treatment of pregnancy on the cicatrix of a previous cesarean section at the uterine isthmus in the first trimester. METHODS: Analysis of 14 patients with pregnancy on the cicatrix of a previous cesarean section at the uterine isthmus in the first trimester was made after conservative treatment by drugs from January 1996 to December 1999. RESULTS: The 14 patients with a pregnancy on the cicatrix of a previous cesarean section at the uterine isthmus in the first trimester were painless, had slight vaginal bleeding, and concurrently had increased serum beta-subunit human chorionic gonadotropin (beta-HCG). Doppler ultrasonic examination revealed an obvious enlargement of the previous cesarean section cicatrix in the uterine isthmus, and found a gestational sac or mixed mass attached to the cicatrice, with a very thin myometrium between the gestational sac and bladder walls. Among the 14 patients, 12 patients had crystalline trichosanthes injected into the cervix, mifepristone taken orally, or methotrexate in the form of intramuscular injection. Following this procedure, their serum beta-HCG dropped to normal. The other 2 patients had a total hysterectomy. CONCLUSIONS: Pregnancy on the cicatrix of a previous cesarean section at the uterine isthmus in the first trimester is a complication of cesarean section. Early diagnosis and effective conservative treatment by drugs are instrumental in decreasing the potential occurrence of uterine rupture, which is also conducive to preserving the patient's future fertility. PMID- 11940342 TI - Correlation of MR imaging and histopathology after partial resection of normal rabbit brain. AB - OBJECTIVES: To investigate the findings of magnetic resonance (MR) imaging and histopathology in early postoperative normal brain, and to define the correlation between MR images and histopathology. METHODS: Thirty-six New Zealand rabbits weighing 2.0 to 3.0 kg were divided into 10 groups according to different postoperative days: 1 to 10 days. A partial resection of the parietooccipital region was performed under usual aseptic conditions after the animals were anesthetized intravenously with 3% pentobarbital (30 mg/kg). MR imaging procedures consisted of pre- and postcontrast scanning and were carried out on postoperative days 1 to 10. Brain tissue samples were prepared for examination immediately after MR scanning. Histopathological examination was done under light both and electron microscopes. The findings of MR imaging were compared with histopathologic findings. RESULTS: Surgical margin contrast enhancement on MR images could be seen 24 hours after surgery. The degree of contrast enhancement increased gradually up to 5 days postoperation, and no remarkable changes were present from days 5 to 10. Disruption of the blood brain barrier (BBB) was the main cause of contrast enhancement during the first 3 postoperative days. After that period, the mechanism responsible for contrast enhancement was the formation of neovascularity and a broken BBB. An increase in the amount of neovascularity played a predominant role in contrast enhancement in normal postoperative brain tissue. CONCLUSIONS: The features of enhanced MR images present at the surgical margin followed a typical time course during the early postoperative period. The role of neovascularity and BBB disruption in the formation of contrast enhancement at the surgical margin varies with time. Knowledge of the features of contrast enhancement in postoperative MR images of normal brain can help in differentiating benign changes from residual malignant glioma. PMID- 11940343 TI - Bone status assessment in Japanese subjects using speed of sound along the tibia. AB - OBJECTIVES: To explore the potential use of ultrasound measurement (SoundScan 2000 Compact, Myriad Ultrasound Systems Ltd., Rehovot, Israel), and to assess both quantitative and qualitative properties of bone. METHODS: The speed of sound waves (SOS; m/s) propagating along the cortical bone was determined at the tibial shaft. The performance, reliability, validity, and clinical application of this system were evaluated in Japanese subjects. RESULTS: In phantom experiments, validity of this system was 0.04 (%), coefficient variation (CV: %) in same-day tests was 0.12, in five-separate-day tests was 0.13, and in independent operators was 0.11, respectively. In experiments with Japanese subjects, CV in same-day tests was 0.38, in five-separate-day tests was 0.99, and in independent operators was 0.90, respectively. In addition, cortical bone status of 1176 Japanese subjects (372 males, 804 females) was clinically evaluated by SoundScan 2000 Compact. SOS was negatively correlated with body fat percentage (%), body fat (kg) and waist hip ratio in females and subjects with higher weight bearing index (leg strength/body weight; WBI) have higher SOS in females under the age of forty. CONCLUSIONS: These findings indicate that the SoundScan 2000 Compact is a highly reliable and valid method for determining cortical bone status and body composition and WBI are closely related to cortical bone status in Japanese females. PMID- 11940344 TI - Deletions are easy detectable in cochlear mitochondrial DNA of Cu/Zn superoxide dismutase gene knockout mice. AB - OBJECTIVES: To investigate the tissue specificity of reactive oxygen species (ROS) damage to mitochondrial DNA (mtDNA) and to determine whether cochlear mtDNA is a sensitive target for ROS damage. METHODS: 10 Cu/ZnSOD gene (Cu/Zn superoxide dismutase gene, Sod1) knockout mice and 16 wild-type mice were analyzed by nested polymerase chain reaction (PCR). RESULTS: Three deletions were detected in various tissues of Sod1 knockout mice. MtDNA3867bp and mtDNA3726bp deletions were the most visible, and mtDNA4236bp deletion was barely detected in these tissues. There were obvious differences in the ratio of deleted mtDNA/total mtDNA in different tissue. Deleted mtDNA was most abundant in the liver and kidney and less in cochlea, heart and brain. The lowest was in spleen and skin. The ratio in various tissues was 3 - 20 times in Sod1 knockout mice over wild-type mice. In cochlea, the ratio was about 15. CONCLUSIONS: Without the protection of Sod1, ROS can lead to mtDNA deletions in various tissues with significant tissue specificity. Cochlear mtDNA is a sensitive target for ROS damage. PMID- 11940345 TI - Possible function of outward potassium currents in isolated Deiters' cells of guinea pig cochlea. AB - OBJECTIVE: To study potassium currents in isolated Deiters' cells of guinea pig cochlea and explore possible function of potassium current in Deiters' cell. METHODS: The whole cell patch clamp recording technique was used to study potassium currents of Deiters' cells in normal external solution and solutions with different K(+) concentrations. We also studied the effects on reversal potentials and outward potassium currents. RESULTS: Isolated Deiters' cells possess voltage dependent, outwardly rectifying ion channels, which are K(+) selective. 50 mmol/L K(+) and 150 mmol/L K(+) in external solution reduced I(K max) from (10.06 +/- 2.2) nA (n = 13) in normal external solution to (6.43 +/- 1.67) nA (n = 6, P < 0.05) and (5.49 +/- 1.33) nA (n = 6, P < 0.05), respectively. While the amplitude of tailcurrents decreased from (468.76 +/- 61.76) pA in 5 mmol K(+) external solution to (224.74 +/- 35.89) pA (P < 0.05) in 50 mmol/L K(+) and to (-911.59 +/- 78.17) pA (P < 0.01) in 150 mmol/L K(+) external solution. CONCLUSIONS: Outwardly rectifying potassium in Deiters' cells could buffer extracellular K(+) in the small space between Deiters' cells and outer hair cells or neural fibers and participate in the diffusion of K(+) from endolymph to perilymph. PMID- 11940346 TI - Expression and effect of basic fibroblast growth factor on human cataract lens epithelial cells. AB - OBJECTIVE: To detect the expression of basic fibroblast growth factor (bFGF) in human ocular tissues, and to assess the effect of bFGF on the proliferation of human cataract lens epithelial cells (LECs) and its correlation with age. METHODS: Enucleated eyes were subjected to immunostaining for bFGF protein. Human cataract LECs were cultured in vitro, and treated with bFGF for 48 hr. Proliferation was estimated by the positive area ratio of proliferating cell nuclear antigen (PCNA) in immunohistochemistry. RESULTS: bFGF protein was found in various human ocular tissues. bFGF stimulated human cataract LEC proliferation, and there was an age-related decrease in responsiveness of human cataract LECs to bFGF (P < 0.05). CONCLUSION: bFGF might play an important role in the proliferation of residual human cataract LECs after cataract surgery. PMID- 11940347 TI - Natural foci of tsutsugamushi disease in the Nan Peng Lie Islands in China. AB - OBJECTIVE: To investigate natural foci of tsutsugamushi disease whose incidence has increased in the Nan Peng Lie Islands in China, an area where this disease has not been previously recorded. METHODS: We recorded the natural foci and isolated Orientia tsutsugamushi (O. tsutsugamushi) organism. We also studied prevention measures. RESULTS: These islands had the natural foci of a south subtropical zone. The main host and vector were Rattus norvegicus and Leptotrombidium deliens (L. deliens), respectively. The seasonal quantity trends of Rattus norvegicus and Leptotrombidium deliens were consistent with the incidence of human infection. Thirty-five strains of O. tsutsugamushi were isolated from Rattus norvegicus and L. deliense. The identification of 7 strains showed that most strains were Karp. Seroepidemiology showed a high prevalence of antibody against O. tsatsugamushi among local people. After prevention measures were used, the incidence was decreased. CONCLUSION: This was the first successful confirmation that the Nan Peng Lie Islands were natural foci of tsutsugamushi disease. PMID- 11940348 TI - Cholecystokinin octapeptide inhibits the in vitro expression of CD14 in rat pulmonary interstitial macrophages induced by lipopolysaccharide. AB - OBJECTIVE: To study the effect of cholecystokinin octapeptide (CCK-8) on lipopolysaccharide (LPS)-stimulated pulmonary interstitial macrophages (PIM) in vitro. METHODS: PIM were isolated and cultured in the presence or absence of LPS, CCK-8, proglumide (the antagonist of CCK receptors) and vehicle. The expression of membrane CD14 (mCD14) protein was assayed by flow cytometry and soluble CD14 (sCD14) in the supernatant was analyzed semi-quantitatively by Western blot. TNF alpha in the supernatant was detected with ELISA. RESULTS: CCK-8, at concentrations of 10(-7) mol/L and 10(-6) mol/L, significantly inhibited the expression of mCD14. Release of sCD14 and TNF-alpha in the supernatant was up regulated by LPS (1 microg/ml) but reduced by CCK-8. The effect of CCK-8 was inhibited by proglumide. CONCLUSION: CCK-8 negatively modulated several functions of LPS-stimulated PIM through CCK receptors. This may be one of the mechanisms for CCK-8 to alleviate inflammation in lung tissue during endotoxemia. PMID- 11940349 TI - Arsenic trioxide (As(2)O(3)) induced apoptosis and its mechanisms in a human esophageal squamous carcinoma cell line. AB - OBJECTIVE: To study whether As(2)O(3) has an apoptotic effect on human solid tumor cells, and the possible cellular and molecular mechanisms of this treatment using human esophageal squamous carcinoma cells (EC8712) as a model. METHODS: DNA microarray, biochemical and cytological analyses were used. RESULTS: The growth and survival of EC8712 cells were markedly inhibited by As(2)O(3) treatment at a concentration of 1, 2 and 4 micromol/L. EC8712 cells were obviously arrested at G2/M phase with As(2)O(3) treatment and apoptosis induced at micromolar As(2)O(3) concentrations, as shown by morphology, histogram related nuclear DNA contents, and DNA gel electrophoresis. As(2)O(3) activated caspase-3, which might be involved in the process of As(2)O(3), induced apoptosis in EC8712 cells. CONCLUSIONS: As(2)O(3) changes the expression of many genes at transcription level. The regulation of expression of many genes might be involved in the process of As(2)O(3) inducing apoptosis. These results suggest that As(2)O(3) can be clinically useful for solid tumor treatment. PMID- 11940350 TI - Lipopolysaccharide upregulates the expression of Toll-like receptor 4 in human vascular endothelial cells. AB - OBJECTIVE: To investigate the expression of Toll-like receptor 2 (TLR2) and Toll like receptor 4 (TLR4) and the effect of lipopolysaccharide (LPS) on their expression in cultured endothelial cells. METHODS: Total RNA was extracted from ECV304 cells and isolated human umbilical vein endothelial cells (HUVECs) exposed to LPS, respectively. The quantification of TLR2 and TLR4 mRNA in HUVECs and EVC304 cells was carried out by reverse transcriptase-polymerase chain reaction (RT-PCR). RESULTS: ECV304 cells and HUVECs were able to express TLR2 and TLR4 mRNA, but the expression levels of TLR4 appeared to be stronger than those of TLR2. LPS could upregulate the expression levels of TLR4 obviously, whereas it had no effect on the expression level of TLR2. CONCLUSIONS: Our data indicate that TLR4 may be the LPS signal transducer in endothelial cells and plays important roles in the cell activation of LPS. The ECV304 cell line is a better experimental model than isolated HUVECs in the research of endothelial cells. PMID- 11940351 TI - Expression of telomerase activity, telomerase RNA component and telomerase catalytic subunit gene in lung cancer. AB - OBJECTIVE: To investigate whether telomerase activity, human telomerase RNA (HTR) and human telomerase reverse transcriptase (HTERT) expression were associated with tumor development in lung cancer and whether telomerase is regulated at gene level or transcriptional level. METHODS: Expression of HTR and HTERT was detected by reverse transcription-polymerase chain reaction (RT-PCR) in 68 human lung cancer and in 68 adjacent-neoplatic lung tissues. And telomerase activity was examined by a quantitative telomeric repeat amplification protocol (TRAP). RESULTS: In 68 lung cancer tissues, telomerase activity, HTR and HTERT were expressed in 79%, 98.5% and 91.2% respectively, whereas all adjacent non neoplastic lung tissues were telomerase negative. Most normal lung tissues expressed HTR (91.2%) and HTERT was detected in only 7 of 68 non-neoplastic tissues. CONCLUSIONS: The relatively high frequency of telomerase activity in lung cancer whereas the detection of no telomerase activity in normal lung tissues suggested that telomerase may play an important role in tumorigenesis of lung cancer. Compared to HTR, HTERT expression was better associated with telomerase activity with a concordance of 88.9%. Telomerase activity may be regulated at transcription level or translation level. PMID- 11940352 TI - Effects of shenmai injection on expression of TNF-alpha mRNA in peritoneal macrophages of scald mice. AB - OBJECTIVE: To explore the effect of shenmai injection (SI) on expression of TNF alpha mRNA in peritoneal macrophages (pMPhis) of scald mice. METHODS: BALB/c mice were inflicted with 11% of body surface area III degree scald and injected intraperitoneally (ip) with SI daily for 5 days, and expression of TNF-alpha mRNA in pMPhis was determined by semi-quantitative RT-PCR. RESULTS: In scald mice, the expression of TNF-alpha mRNA in pMPhis increased significantly, but it was reduced obviously (P < 0.01) after SI administration, while the livability was increased markedly (P < 0.05). CONCLUSIONS: For scald mice, the cause of death at early stage might be related to the high expression of TNF-alpha mRNA in pMPhis and the use of SI can decrease the death rate. PMID- 11940353 TI - Studies on the DNA content of breast carcinoma cells with neuroendocrine differentiation. AB - OBJECTIVE: To make quantitative analysis of DNA content of breast cancer with neuroendocrine (NE) cells and its significance. METHODS: Using MIPS-III image analyzer, DNA content and 9 parameter measurements of the tumor nuclei were made in both NE positive (17) and negative (64) breast carcinomas. RESULTS: Out of 81 breast carcinomas, 17 cases were NE positive while 64 cases were NE negative. In the NE (+) cases, the integral optic density, mean optic density, DNA index, DNA stemlines peak, > 5c aneuploidy cells and the rate of aneuploidy cells were all lower than those in the NE negative breast carcinoma cases (P < 0.01). The positive rates of NE cells were 32.5% and 7.9% in grade I - II breast carcinomas and in grade III breast carcinomas respectively with significant difference between the two groups (P < 0.01). CONCLUSION: Our study shows that NE (+) breast carcinomas have lower DNA parameters than NE (-) breast carcinomas, suggesting that NE (+) breast carcinomas have lower malignancy. PMID- 11940354 TI - A new method for internal cardioversion in patients with persistent atrial fibrillation. PMID- 11940355 TI - Effect of necrotic tissue on progressive injury in deep partial thickness burn wounds. AB - OBJECTIVE: To evaluate the influence of necrotic tissue on progressive injury in deep partial thickness burn wounds. METHODS: Tissue specimens were cultured both for estimation of IL-8, EGF, bFGF, PDGF-AB and histopathological examination, from the pre-operation, post-operation, and non-operation wounds from seven patients with deep partial thickness burn. RESULTS: In seven specimens from the non-operation group, IL-8 release increased compared with those in the post operation group (P < 0.001), while the levels of EGF, bFGF, PDGF-AB release were lower than those in the post-operation group. Histopathological examination revealed that in the non-operation group, the degree of neutrophil infiltration was enhanced, the extent of tissue necrosis enlarged, and residual skin appendages disappeared. In contrast, in the post-operation group, the degree of inflammatory response was decreased, with the formation of fresh granulation tissue and epithelialization. CONCLUSION: This study suggests that the presence of necrotic tissue could be the inhibitory factor in the wound healing process, as it might cause tissue progressive injury leading to the delay of wound healing. To promote wound healing, active tangential excision is recommended to remove necrotic tissue. PMID- 11940356 TI - Expression of positive and negative regulators of cell cycle during wound healing. AB - OBJECTIVE: To detect the expression of cell cycle positive regulators cyclin D(1), cyclin E, CDK(2), CDK(4) and negative regulators p21(cip1), p27(kip1), p16(ink4a) and p15(ink4b) during wound healing in rats. METHODS: Open wounds of full-thickness skin, diameter 1.8 cm, on rat backs were used as the wound model. Wound tissues were harvested on postwounding days 3, 5, 7, 9, 11, 14, 21 and 30. Ki67 expression in granulation tissue was detected by immunohistochemical assay. The patterns of the expression of cyclin D(1), cyclin E, CDK(2), CDK(4) and p21(cip1), p27(kip1), p16(ink4a), p15(ink4b) were detected by Western blot. RESULTS: Cell proliferation in granulation tissue took place predominantly within the first week after injury, with the proliferation peak occurring at postwounding day 5. There were no dramatic variations in the expression of cyclin D(1), CDK(2) and CDK(4) during wound healing. Up-regulated cyclin E was maintained from day 3 to 11 after injury, and then was down-regulated. No expression of p16(ink4a) and p15(ink4b) was found. p21(cip1) was expressed only from day 7 to 14, with peak expression observed on day 9. Constitutive p27(kip1) was expressed throughout wound healing with low levels in the proliferating period of day 3 to 5 and with increased levels in the post-mitotic and remodeling stage. The expression of p21(cip1) and p27(kip1) showed an inverse gradient to that of Ki67. CONCLUSION: p21(cip1) and p27(kip1) play a supervising role in preventing the hyperproliferative tendency in tissue repair. PMID- 11940357 TI - Healing of chronic cutaneous wounds by topical treatment with basic fibroblast growth factor. AB - OBJECTIVE: To evaluate the safety and efficacy of topical application of recombinant bovine basic fibroblast growth factor (rbFGF) on the healing of chronic cutaneous wounds. METHODS: Twenty-eight patients with thirty-three chronic cutaneous wounds resulting from trauma, diabetes mellitus, pressure sore and radiation injuries were enrolled in this prospective, open-label crossover trial. Prior to treatment with rbFGF, all wounds failed to heal with conventional therapies within 4 weeks. All wounds were locally treated with rbFGF at a dose of 150 AU/cm(2). Healing time and the quality of wounds were used to evaluate the efficacy of the treatment. RESULTS: Healing of all chronic wounds was expedited. During the study, eighteen wounds completely healed within 2 weeks, four healed within 3 weeks, and another eight completely healed within 4 weeks. Only three wounds failed to heal within 4 weeks, but healed at 30, 40 and 42 days after treatment with rbFGF. Thus, compared with conventional therapies, the effective rate of rbFGF treatment within 4 weeks was 90.9%. Histological assessment showed more abundant capillary sprouts or tubes and that fibroblasts were differentiated in wounds treated with rbFGF. No adverse side effects related to basic fibroblast growth factor were observed. CONCLUSIONS: Our results indicate that rbFGF could be used to accelerate healing in chronic wounds. It is our belief that this may be a more effective method of chronic wound management. PMID- 11940358 TI - Mechanism of thoracolumbar burst fractures: a biomechanical study. AB - OBJECTIVE: To investigate the mechanism associated with thoracolumbar burst fractures. METHODS: Stress distribution of the spine in the upright, flexion and extension positions was analyzed using a three-dimensional finite element model of the spinal motion segment. RESULTS: Stress concentration was noted at the pedicle and posterosuperior part of the vertebral body near the pedicle. CONCLUSION: Stress concentration of the spine may be implicated in the biomechanical mechanism underlying thoracolumbar burst fractures. PMID- 11940359 TI - Nestin expression and proliferation of ependymal cells in adult rat spinal cord after injury. AB - OBJECTIVE: To determine cell proliferation and nestin expression in the ependyma of adult rat spinal cord after injury. METHODS: Rat spinal cord injury models were established by aneurysm clip compression, and nestin expression and proliferation of ependymal cells at different times were shown with pathological and immuno-histochemical staining. RESULTS: Ependymal cells adjacent to the injured site demonstrated a dramatic increase in nestin expression 24 hours after compression. Proliferating cell nuclear antigen was positive, and significant proliferation was observed after 7 days. Nestin expression was down regulated as time went by. CONCLUSION: Normally quiescent mature ependymal cells appear to revert to an embryonic state in response to spinal cord injury. PMID- 11940360 TI - Management of cerebral ischemia due to Takayasu's arteritis. AB - OBJECTIVE: To explore the management of cerebral ischemia caused by Takayasu's arteritis. METHODS: Ninety-three cases treated from June 1984 to September 1999 at the General Post & Telecom Hospital, the Sir Run Run Shaw Hospital, the First Affiliated Hospital of Zhejiang University, the Second Medical College of Beijing University, Beijing An Zhen Hospital, and the Beijing Union Medical College Hospital, including 10 men and 83 women, were reviewed. Of the 93 cases, bypasses from the ascending aorta to the axillary or subclavian artery and from graft to the carotid artery were performed in 47 cases. Subclavian to carotid bypass was performed in six cases. Percutaneous transluminal angioplasty (PTA) was used in five cases and stenting in one. RESULTS: Marked improvement was achieved in 30.3%, fair in 34.9%, improvement in 21.2%, unchanged in 4.6%, and death in 9.0% before discharge; 30.6%, 38.8%, 16.3%, 4.1%, and 2.0% respectively during a mean follow-up of 48 months, and recurrence requiring revision in 8.2%. CONCLUSION: Patients with occlusive lesions of all four cervical arteries always have severe cerebral ischemia and their distal runoff is always unvisualised by angiography. However, we found by exploration that the internal carotid artery is patent in all but one patient. Therefore, an ascending aorta to carotid bypass is feasible in most instances, and this can and should be done when the cerebral perfusion is jeopardized at a time when the patient is in a stable or relatively stable condition. Unfortunately, the cerebral re-perfusion syndrome is still a serious and not completely solved problem. PMID- 11940361 TI - Comparison of three doses of enalapril in preventing left ventricular remodeling after acute myocardial infarction in the rat. AB - OBJECTIVE: To compare the effects of high, middle and low doses of enalapril in preventing left ventricular remodeling (LVRM) after acute myocardial infarction (AMI) in rats, especially evaluating the efficacy of low dose enalapril. METHODS: AMI was induced by ligating the left coronary artery in 149 female SD rats. 48 hours after the procedure, the 97 surviving rats were randomized to one of the following four groups: (1) AMI controls (n = 24), (2) high-dose (10 mg x kg(-1) x d(-1), n = 25), (3) middle-dose (1 mg x kg(-1) x d(-1), n = 23), and (4) low-dose (0.1 mg x kg(-1) x d(-1), n = 25) enalapril groups. In addition, sham-operated (n = 13) and normal rats (n = 10) were randomly selected to serve as non-infarction controls. Enalapril was delivered by direct gastric gavage. After 4 weeks of therapy, hemodynamic studies were performed, then the rat hearts were fixed with 10% formalin and pathology analysis was performed. Exclusive of the dead rats and those with MI size < 35% or > 55%, complete experimental data were obtained from 67 rats, which were comprised of (1) AMI controls (n = 13), (2) high-dose enalapril (n = 13), (3) middle-dose enalapril (n = 12), (4) low-dose enalapril (n = 12), (5) sham-operated (n = 8) and (6) normal (n = 9) groups. RESULTS: There were no significant differences among the four AMI groups in infarction size (all P > 0.05). Compared with the sham-operated group, the left ventricular (LV) end diastolic pressure (LVEDP), volume (LVV), absolute and relative weight (LVAW, LVRW) in AMI group were all significantly increased (all P < 0.001), while maximum LV pressure rising and dropping rates (+/- dp/dt) and their corrected values by LV systolic pressure (+/- dp/dt/LVSP) were all significantly reduced in the AMI control group (P < 0.01 - 0.001), indicating LVRM occurred and LV systolic and diastolic functions were impaired. Compared with the AMI group, LVEDP, LVV, LVAW and LVRW were all significantly decreased in the three enalapril groups (control P < 0.001), with the reduction of LVEDP, LVV and LVAW being more significant in high-dose than in low-dose enalapril groups (all P < 0.05), and the +/- dp/dt/LVSP were significantly increased only in the high and middle-dose enalapril groups (P < 0.01). CONCLUSIONS: High, middle and low doses of enalapril were all effective in preventing LVRM after AMI in the rat, with low dose enalapril being effective and high dose superior. As for LV functional improvement, only high and middle-dose enalapril were effective. PMID- 11940362 TI - Dynamic changes in type I collagen, MMP-1 and TIMP-1 after angioplasty. AB - OBJECTIVE: To investigate the dynamic changes of type I collagen, and the activity of metalloproteinases-1 (MMP-1) and tissue inhibitor of metalloproteinases-1 (TIMP-1) after angioplasty. METHODS: The restenotic model of iliac arteries of domestic microswine was established with hypercholesterol feed plus two angioplasties. Angioplastied vessels were harvested at the end of 1, 2, 3 and 6 months after the second angioplasty. Immunohistochemistry, transmission electronic microscopy and image quantitative analysis techniques were employed to study neointimal proliferation, the phenotype of vascular smooth muscle cells (VSMC) and the expression of type I collagen, MMP-1 and TIMP-1. RESULTS: The peak of vascular neointimal proliferation was at 3 months after angioplasty. The expression of type I collagen gradually increased from 1 to 6 months after angioplasty. For MMP-1, expression was lower in the early stage after angioplasty but increase to normal levels of control vessels at 6 months after angioplasty. Expression of TIMP-1 rapidly increased in the early phase after angioplasty, reached peak at 3 months and maintained the high level till 6 months after angioplasty. Meanwhile, the VSMC was predominantly the synthetic phenotype at the early stage and was transformed to the contractive phenotype at the late stage after angioplasty. The ratio of TIMP-1 and MMP-1 was positively related to the area of the neointima and the expression of type I collagen respectively (P < 0.01). CONCLUSION: Type I collagen increased gradually after angioplasty, which might be determined by the ratio of TIMP-1/MMP-1 and also related to the phenotype of VSMC. PMID- 11940363 TI - Stent supported coronary angioplasty in patients with severe ventricular dysfunction. AB - OBJECTIVE: To assess the immediate- and long-term outcomes of stent supported coronary angioplasty in patients with severe left ventricular dysfunction. METHODS: Seventy-four consecutive patients with angiographic left ventricular ejection fractions < or = 40% underwent percutaneous coronary angioplasty, including 61 (81%) with stent back-up. Clinical follow-up (29.1 +/- 22.9 months) data were obtained in all clinically successful patients. RESULTS: Angiographic success was achieved in 120 (89.5%) of 134 lesions. Clinical success was obtained in 66 (89.2%) patients with 1.4% procedure related mortality, 2.7% myocardial infarction and zero need of emergent bypass surgery. NYHA class reduction > or = 1 was seen in 29 (73%) of 40 successfully treated congestive heart failure patients at 6 months after the procedure. During long-term follow-up, 58 (87.9%) of 66 patients with clinical success were alive, including 44 (68.6%) free from cardiac events. CONCLUSIONS: Patients with severe left ventricular dysfunction treated with stent supported PTCA experience a high rate of success, low procedure related mortality and satisfactory long-term survival. PMID- 11940364 TI - Pump models assessed by transesophageal echocardiography during cardiopulmonary resuscitation. AB - OBJECTIVE: Transesophageal echocardiography was performed during closed-chest cardiopulmonary resuscitation (CPR) in in-hospital cardiac arrest to further explore the hemodynamic mechanism of CPR. METHODS: CPR attempts were performed according to advanced cardiovascular life support guidelines in 6 cases of in hospital cardiac arrest. Multi-plane transesophageal echocardiography was carried out within 15 min of initiation of CPR. Throughout CPR, the motion of the mitral, tricuspid and aortic valves, the changes in the left ventricular cavity size and the thoracic aortic diameter were observed. Trans-mitral and trans-aortic Doppler files of blood flow were also documented. RESULTS: A closure of the mitral and tricuspid valves with simultaneous opening of the aortic valve occurred exclusively during chest compression, resulting in forward blood flow in the pulmonary and systemic circulation. Peak forward aortic flow at a velocity of 58.8 +/- 11.6 cm/s was recorded during the compression phase. Whereas, a closure of the aortic valve and rapid opening of the atrioventricular valves associated with ventricular filling during relaxation of chest compression was noted in all 6 patients. Peak forward mitral flow at a velocity of 60.6 +/- 20.0 cm/s was recorded during the release phase. Mitral regurgitation during the chest compression period was detected in 5 patients, reflecting a positive ventricular to-atrial pressure gradient. A reduction in the left ventricular chamber and an increase in the thoracic aortic diameter during the compression phase was found in all patients, indicating that direct cardiac compression contributed to forward blood flow. CONCLUSION: These observations favor the cardiac pump theory as the predominant hemodynamic mechanism of forward blood flow during CPR in human beings. PMID- 11940365 TI - Influence of Valsartan on myocardial apoptosis in spontaneously hypertensive rats. AB - OBJECTIVE: To explore the pathogenic changes of myocardial apoptosis in heart hypertrophy during hypertension and evaluate the anti-apoptosis effect of Valsartan. METHODS: Thirty spontaneously hypertensive rats (SHRs) were divided into two groups: 15 treated with Valsartan (20 mg x kg(-1) x d(-1)) (SHR + Valsartan group), the others with placebo (SHR + placebo group), with 15 normal Wistar rats as control. Systolic blood pressure was measured by the tail-cuff method. The observation period was from 8 to 16 weeks of age. Cardiac apoptosis was evaluated by a Terminal Deoxynucleotidyl Transferase-Mediated dUTP-biotin Nick End Labeling (TUNEL) assay. RESULTS: Mean blood pressure values were 127 +/- 2 mm Hg in controls, 163 +/- 6 mm Hg in the SHR + Valsartan group and 193 +/- 7 mm Hg in the SHR + placebo group at 16 weeks of age, whereas the blood pressure in 8-week-old SHR and Wistar rats were 175 +/- 3 mm Hg and 125 +/- 5 mm Hg, respectively. The ratio of the heart weight over body weight declined in Wistar (3.07 +/- 0.03 mg/g) and SHR + Valsartan groups (3.22 +/- 0.19 mg/g) compared with the SHR + placebo group (4.02 +/- 0.31 mg/g) (P < 0.05). The density of TUNEL-positive cells in Wistar and SHR +/- Valsartan groups was 23.3 +/- 3.3 nuclei/HPF and 35.0 +/- 1.3 nuclei/HPF, both of which were significantly less than that of the SHR + placebo group (116.7 +/- 11.3 nuclei/HPF). CONCLUSIONS: In response to chronic pressure overload, cardiomyocyte-specific apoptosis contributes to the transition from compensatory hypertrophy to decompensation. Apoptosis may be effectively inhibited by Valsartan in the early stage of hypertension. PMID- 11940366 TI - Association of the phosphatidylinositol signal pathway with prolonged myocardial ischemia. AB - OBJECTIVE: To study the changes in activity of phosphatidylinositol 4 kinase (PI 4 kinase), phosphatidylinositol 4 phosphate 5 kinase (PIP 5 kinase) and protein kinase C (PKC) during myocardial ischemia and elucidate the relationship between phosphatidylinositol signal pathways and prolonged myocardial ischemia. METHODS: In vivo an ischemic rat model was used. Activity of PI 4 kinase, PIP 5 kinase and PKC were measured at different times in postischemic heart cells using isotope analysis. RESULTS: The activity of PI kinase, PIP kinase and PKC in the myocardium increased to peak at 1 hour postischemia, with activities 6.1, 3.0 and 4.0 fold over control levels, respectively. Their activities declined to normal levels with time. CONCLUSION: The phosphatidylinositol signal pathway is involved in prolonged myocardial ischemia, but its mechanism needs further study. PMID- 11940367 TI - Cholesteryl ester transfer protein levels and gene deficiency in Chinese patients with cardio-cerebrovascular diseases. AB - OBJECTIVE: To detect cholesteryl ester transfer protein (CETP) levels, frequencies of CETP D442G and I 14A mutations and characteristics of abnormal lipids in patients with cardio-cerebro vascular diseases. METHODS: Ninety-four myocardial infarction (MI) patients, 110 stroke patients and 335 healthy controls were selected. The CETP concentration was determined using ELISA. The CETP activity was measured using a substrate of (14)C-radiolabeled discoidal bilayer particles. The CETP gene mutations were detected by PCR-RFLP. RESULTS: The CETP concentrations in the MI and stroke group, were higher than those in the controls. The gene mutation frequencies of D442G in the MI, stroke and control group were 3.5%, 3.6% and 5%, respectively, and the frequencies of I 14A were 1.05%, 0.91% and 1%, respectively. One case of D442G homozygote was detected in the healthy group. The frequency of two CETP gene mutations showed no significant difference among the patients and controls. The CETP concentration and activity in subjects with CETP mutations were one-third of those in the control group. The level of HDL-C, apo-A1 increased in the mutation subjects, while the TG level decreased. CONCLUSIONS: The CETP level increased significantly in patients with cardio-cerebrovascular diseases. The carriers of CETP deficiency had CETP and lipid abnormalities. PMID- 11940368 TI - Ventricular remodeling by Scutellarein treatment in spontaneously hypertensive rats. AB - OBJECTIVE: To observe reversal of ventricular remodeling by the protein kinase C inhibitor Scutellarein in spontaneously hypertensive rats (SHRs). METHODS: Twelve SHRs were randomly divided into two groups. Scutellarein and saline (10 mg x kg( 1) x d(-1)) were given by intraperitoneal injection to two groups of rats separately. Systolic blood pressure (SBP) and ventricular weight index (LVW/BW, RVW/BW) were measured. A polarization microscope and an image analyzer system (IAS) were used to observe changes in cardiovascular structure and to count the content of cardiac muscle interstitial collagen. RESULTS: The pathologic changes in the left ventricle in the Scutellarein group rats (SHR(D)) improved to varying degrees, including hypertrophy of the cardiac muscle and collagen volume fraction. CONCLUSION: Scutellarein can reverse ventricular remodeling, improve myocardial stiffness and protect heart cardiac muscle. PMID- 11940369 TI - Effect of resveratrol on platelet aggregation in vivo and in vitro. AB - OBJECTIVE: Low or moderate consumption of red wine has a greater benefit than the consumption of other beverages in the prevention of atherosclerosis and coronary heart disease and this is increasingly attributed to the polyphenol compounds in red wine, such as resveratrol. In the present study, we investigated the effect of resveratrol on platelet aggregation in vitro and in vivo. METHODS: Platelet aggregation in rabbits and normal subjects was measured using Born's method. RESULTS: Resveratrol, at 10 - 1000 micromol/L, significantly inhibited platelet aggregation in vitro induced by collagen, thrombin, and ADP in healthy subjects. The inhibitory effect was concentration-dependent. Hypercholesterolemia induced by high-cholesterol diet enhanced ADP-induced platelet aggregation. Resveratrol 4 mg x kg(-1) x d(-1) inhibited ADP-induced platelet aggregation in vivo despite no changes in serum lipid levels. CONCLUSIONS: Resveratrol inhibits platelet aggregation both in vitro and in vivo. This may be one of the mechanisms by which resveratrol prevents atherosclerosis. PMID- 11940370 TI - Helicobacter pylori antigen and its IgG, IgA -type specific immunocomplexes in sera from patients with Helicobacter pylori infection. AB - OBJECTIVE: To explore the characteristics of Helicobacter pylori (H. pylori) antigen in serum and to evaluate its clinical diagnostic value. METHODS: Enzyme linked immunosorbant assay (ELISA) was developed to detect the soluble H. pylori antigen (S-Hp) and circulatory specific H. pylori antigen immunocomplexes (Hp-IC) in serum. RESULTS: The positive rate of S-Hp was 90.91% from 66 patients with H. pylori infection, which was much greater than 0% found in 28 controls (P < 0.001). Moreover, its concentration closely reflected the number of H. pylori in the gastric mucosa layer. We also found that Hp-IC existed bound with IgG and/or IgA in patients with positive S-Hp. However, there is no evidence to show the concentration of S-Hp reduced significantly in followed-up subjects after effective therapy. CONCLUSIONS: These methods as newly and noninvasive complementary tools can be used for clinical diagnosis of H. pylori infection. In addition, S-Hp and Hp-IC may be of importance in H. pylori pathogenesis. PMID- 11940371 TI - Role of the area postrema of medulla oblongata in the regulation of canine interdigestive migrating motor complex. AB - OBJECTIVE: To determine the effect of the area postrema (AP) of the medulla oblongata on gastrointestinal interdigestive migrating motor complex (MMC) and the plasma motilin level. METHODS: Interdigestive MMC activities of the antrum and duodenum were recorded by strain gauge implanted on the serosa of 8 conscious dogs. A cannula was intubated in femoral vein for motilin injection. The plasma motilin concentration was measured by RIA. We observed: (1) normal interdigestive MMC activity and fluctuations in plasma motilin concentration; (2) the effects of electrically damaging the AP on MMC activity and plasma motilin level; (3) whether intravenous injection of motilin could induce phase III contractions after the AP was destroyed. RESULTS: (1) Typical interdigestive MMC with phase I, II, III, and IV was recorded in normal dogs. Phase III was concurrent with the peak of plasma motilin level. (2) In damaged AP dogs, antroduodenal interdigestive MMC contractions were suppressed; cyclic, phasic and migratory pattern of MMC was disrupted. Plasma motilin concentration was decreased. Intravenous injection of motilin could not induce phase III contractions. CONCLUSIONS: The area postrema might play an important role, which is mediated by motilin, on the regulation of interdigestive MMC. PMID- 11940372 TI - Urinary type IV collagen: a specific indicator of incipient diabetic nephropathy. AB - OBJECTIVE: To determine whether urinary type IV collagen can serve as an indicator specific for diabetic nephropathy. METHODS: Using a novel sandwich ABC ELISA to measure type IV collagen directly, the 24-hour urinary type IV collagen excretion rate was determined in 120 diabetic patients and some groups of controls. Urinary albumin determinations were made with a RIA kit at the same time. A total of 13 diabetic patients with microalbuminuria underwent percutaneous renal biopsy for definitive diagnosis of diabetic nephropathy. Type IV collagen and TGF-beta 1 immunoreactivities were detected with ABC methods in renal biopsies. RESULTS: Urinary type IV collagen excretion was significantly increased in diabetic patients with microalbuminuria, especially those with albumin excretion above 200 mg/24 h. By comparison, collagen excretion was equivalent to that in healthy controls when measured in diabetics with normalbuminuria and in patients with primary glomerular disease, primary hypertension, or coronary heart disease. Urinary type IV collagen excretion in diabetics was negatively correlated with creatinine clearance. In renal biopsies from subjects with elevated collagen excretion, the glomeruli showed pathological changes typical of diabetic nephropathy. Also, excessive type IV collagen and TGF beta 1 immunoreactivity were detected in the glomeruli, Bowman's capsule and interstitium. CONCLUSIONS: Excretion of type IV collagen, possibly reflecting increased production or decreased degradation of this protein, may be a clinically useful indicator of incipient diabetic nephropathy. PMID- 11940373 TI - Deep cerebral venous thrombosis in adults. AB - OBJECTIVE: To investigate the pathogenesis, clinical features, radiographic findings and therapeutic outcomes of non-acute intracranial deep venous thrombosis in adults. METHODS: Five patients who presented with increased intracranial pressure were examined with computed tomography, magnetic resonance and angiography, diagnosed as having non-acute intracranial deep venous thrombosis, and treated with thrombolytic therapy. They were reviewed retrospectively. RESULTS: There were 3 men and 2 women, aged from 22 to 49 years. Symptom duration ranged from 1 month to 7 months, and 4 of the 5 patients were associated with venous sinus thrombosis. Two patients developed cold and fever before the onset of disease, and 3 patients had no evident predisposing factors. After the infusion of thrombolytic and systemic anti-coagulant therapy, the neurological symptoms and signs of the patients were alleviated. CONCLUSIONS: Digital subtraction angiography (DSA) is more sensitive and accurate than MRI on diagnosing intracranial deep venous thrombosis. It may play an important role in the assessment of the treatment of intracranial deep venous thrombosis. Thrombolysis and anticoagulation of intracranial deep venous thrombosis appears to be a safe and efficacious treatment not only in the acute stage but also in the non-acute stage. PMID- 11940374 TI - Clinical analysis of 78 cases of paroxysmal nocturnal hemoglobinuria diagnosed in the past ten years. AB - OBJECTIVE: To learn more about the clinical and laboratory features of patients with paroxysmal nocturnal hemoglobinuria (PNH) diagnosed in the past ten years. METHODS: Clinical and laboratory data for 78 cases of PNH diagnosed from January 1990 to November 1999 in our hospital were analyzed retrospectively. RESULTS: In comparison with PNH cases reported in the 1980s, the newly diagnosed PNH cases showed the following features: (1) older age of disease onset (from 27 to 34 years); more female cases (from 18.5% to 38.5%); more cases without hemoglobinuria (from 24.2% to 38.5%). (2) No positive family hereditary history. (3) Bone marrow dysplasia, abnormal karyotype and negative sister chromatid differentiation were found in 19.2%, 12.2% and 8.9% of the PNH patients, respectively. 12.3% of the patients had bone marrow hypoplasia, and most of them had no hemoglobinuria. Ham's tests were negative in about 34.2% of the cases. CD55 and CD59 on peripheral blood cells were deficient in 100.0% of the cases, suggesting that CD55 and CD59 tests can improve the diagnosis of PNH. (4) Adrenocortical hormone was effective in 83.8% of the patients, 54.2% of whom relapsed within one year. Eight refractory and relapsed patients were treated with low dose chemotherapy (MP therapy: Melphalan 2 - 6 mg x d(-1); Prednisone 0.5 mg x kg(-1) x d(-1)). Five (62.5%) of them showed positive responses. Bone marrow failure and other side effects were not serious in this group of patients. CONCLUSIONS: PNH, an acquired blood disease seen more often among adult males, can be diagnosed more sensitively by hemocyte member CD55 and CD59 tests and treated more effectively with adrenocortical hormone or low dose chemotherapy. PMID- 11940376 TI - Early and late outcomes in patients with severe extracranial internal carotid stenosis undergoing carotid endarterectomy. AB - OBJECTIVE: To determine the perioperative and late outcomes for carotid endarterectomy (CEA) in treatment of patients with high-grade stenotic lesions of the extracranial internal carotid artery. METHODS: Twenty patients underwent 21 CEAs at the Zhongshan Hospital between May 1993 and June 2000. They were 19 men and 1 woman, with a mean age of 64 +/- 9 years. Seven patients were performed for stroke, 11 for transient ischaemic attacks (TIAs) and 2 for asymptomatic disease. Duplex scan was the primary tool of evaluation prior to surgery. Perioperative digital subtraction angiography and magnetic resonance angiography were done for 19 and 18 patients, respectively. The percentage of stenosis was calculated using NASCET criteria. Of the 21 lesions operated, 19 had a stenosis of 70% or greater, 2 had ulcerative lesions with a stenosis ranging from 60% to 69%. All CEAs were performed under cervical block anaesthesia with selective intraoperative shunting and patch angioplasty. The patients were followed up regularly with duplex scan surveillance. RESULTS: There was no mortality or stroke during 30 days postoperatively. A TIA occurred in one patient and cranial nerve injury in 2 patients perioperatively. All patients were followed up for a mean interval of 31 +/- 20 months (range: 1 - 63 months). The 2-year survival rate and risk of stroke were 92.3% and 0%, respectively, and the 5-year survival rate and risk of stroke were 79.1% and 12.5%, respectively. Two asymptomatic recurrent stenoses ranging from 50% to 60% were detected on follow-up duplex scan. CONCLUSIONS: For the patients in this study, CEA is associated with an acceptable perioperative outcome as well as a satisfactory long-term beneficial effect in stroke prevention. PMID- 11940375 TI - Treatment of osteoporosis in men using dehydroepiandrosterone sulfate. AB - OBJECTIVE: To study the effect of dehydroepiandrosterone sulfate (DHEAS) treatment of osteoporosis in men with T(BMD) > or = 2.5SD. METHODS: Eighty-six patients were randomly divided into two groups: treatment group (n = 44) and control group (n = 42). DHEAS (100 mg q.d.) was given to the treatment group for 6 months. Bone mineral density, (BMD), biochemical markers of bone absorption and formation and other serum biochemical markers were measured before and after DHEAS treatment. Drug side effects were also evaluated. RESULTS: After oral administration of DHEAS (100 mg q.d.) for 6 months, the serum concentrations of DHEAS and IGF-I in the treatment group were 93.75% +/- 16.1% and 17.71% +/- 4.2% higher respectively than those in the control group (P < 0.01). The BMD of L2, L3, L4, L2 - 4 and Neck sections increased in the treatment group by 2.65% +/- 0.65%, 2.70% +/- 0.48%, 3.10% +/- 0.41%, 2.82% +/- 0.37% and 2.32% +/- 0.31%, respectively, as compared with that the control group (P < 0.05 or 0.01). No significant changes were observed in serum FT, E(2) and PSA concentrations in the treatment group as compared with the control group. CONCLUSION: The treatment of osteoporosis in men with DHEAS is safe and effective. PMID- 11940377 TI - Adenovirus-mediated TGF-beta(1) gene transfer to human degenerative lumbar intervertebral disc cells. AB - OBJECTIVE: To test the possibility of modification of human degenerative lumbar disc cells by the exogenous growth factor gene, transforming growth factor beta(1) (TGF-beta(1)) cDNA, and the expression of the encoded protein. METHODS: Nucleus pulposus samples were surgically obtained from 8 patients with degenerative lumbar disc disease. The cells were cultured and directly infected by two adenoviral constructs, Ad/CMV-EGFP containing the enhanced green fluorecence protein (EGFP) gene (marker gene) and Ad/CMV-TGF-beta(1) containing the potentially therapeutic TGF-beta(1) gene. Transgene expression was analyzed by fluorescence production and immunohistochemical staining (Ad/CMV-TGF-beta(1)). RESULTS: Culture cells transduced by Ad/CMV-EGFP showed specific green fluorescence under the fluoroscope and expression sustained for at least 4 weeks. When infected by Ad/CMV-TGF-beta(1), approximately 30% of cultured cells were stained brown (+) with TGF-beta(1) staining. CONCLUSION: This study established the strategy of delivering a potentially therapeutic gene, TGF-beta(1), by using an adenoviral vector to human degenerative lumbar intervertebral disc cells. PMID- 11940378 TI - Quantitative evaluation of the anal sphincter by the tendency presented in the manometric asymmetry variations. AB - OBJECTIVES: To assess the tendency of manometric asymmetry, and to quantitatively evaluate the function of the anal sphincter. METHODS: Using the PC polygram HR at the states of relax and squeeze, we compared the values of pressure asymmetry in the anorectum of 27 normal children with that of 12 children with fecal incontinence. RESULTS: In the two states, the anal sphincter in normal children showed not only the gradient of pressure, but also a gradient of pressure asymmetry. Children with fecal incontinence did not show any gradients. CONCLUSION: Gradients of pressure and pressure asymmetry can quantitatively evaluate the function of the anal sphincter. PMID- 11940379 TI - Overexpression of type I growth factor receptors in pterygium. AB - OBJECTIVE: To evaluate the expression of the type I growth factor receptor family [epidermal growth factor receptor (EGFR), ErbB2 and ErbB3] by the epithelial cells in pterygium. METHODS: Immunoflourescent staining and Western blotting were performed to detect the expression pattern and quantity of EGFR, ErbB2 and ErbB3 proteins in the epithelia of 15 patients with primary pterygium and 12 subjects with normal conjunctiva. RESULTS: In immunofluorescent staining, the EGFR protein was present in the basal cells while the ErbB2 and ErbB3 were expressed by the superficial cells in normal conjunctival epithelium. Of the pterygium cases 15, 11 were stained by EGFR, ErbB2 and ErbB3 in the full thickness of the epithelium and showed stronger staining compared with the control group. Four of them showed a similar staining pattern to the normal conjunctiva group. The density of protein bands detected by Western blotting for all three growth factor receptors was consistent with the immunofluorescent staining. Compared with normal conjunctiva, stronger protein bands of these three receptors were found in all of the pterygium specimens, in which EGFR, ErbB2 and ErbB3 were expressed in the full thickness, as shown by immunofluorescent staining. CONCLUSIONS: The increased expression of EGFR, ErbB2 and ErbB3 proteins was present in pterygium, which indicated that pterygium is a disorder with abnormal proliferation. The abnormal expression of EGFR, ErbB2 and ErbB3 by the epithelium and the communication with cytokines in the stroma in pterygium may be a key pathogenic factor in this disorder. PMID- 11940380 TI - T helper cells in patients with chronic hepatitis B virus infection. AB - OBJECTIVE: To investigate the compositions of Th1/Th2/Th3 cells in chronic hepatitis B virus (HBV)-infected individuals by determining the expression of interleukin-4 (IL-4), inetrferon-gamma (IFN-gamma), and transform growth factor beta (TGF-beta) in single CD4(+) T cells isolated from peripheral blood mononuclear cells (PBMCs) and the role of polarized Th cell populations in chronic HBV-infection was discussed. METHODS: PBMCs from chronically infected HBV individuals were isolated, stimulated by PMA/Ionomycin/Monensin, and IL-4, IFN gamma and TGF-beta production by CD4(+) T cells was determined by using fluorescence activated cell sorter (FACS) analysis. RESULTS: The percentage of IFN-gamma-producing T cells, IL-4-producing T cells and TGF-beta-producing T cells ranged from 2.3% - 18.6%, 1.1% - 8.7% and 0.7% - 7.1% respectively in CD4(+) T cells from non-infected individuals. Most of CD4(+) T cells from PBMCs in chronically infected HBV individuals were Th0 cells. The proportion of Th1 cells increased significantly with hepatic inflammatory activity, and in the active period of chronic hepatitis B infection were higher than those in the non active period (P < 0.05). Th2 cell percentage in CD4(+) T cells from HBV-infected individuals did not differ significantly (P > 0.05), but were higher than that from controls (P < 0.05). Th3 cell percentage in CD4(+) T cells from asymptomatic carrier (AsC) group was higher than that in the chronic hepatitis B (CHB) and control groups (P < 0.05). CONCLUSIONS: Th1 phenotype cytokines were positively correlated with hepatic inflammatory activity in chronic hepatitis B and Th2 cells may be associated with the persistence of HBV infection. Th3 cells cooperating with Th2 cells can negatively regulate immune responses and may be associated with the immune tolerant state of chronic HBV infection. PMID- 11940381 TI - Effects of antisense glutamic acid decarboxylase oligodeoxynucleotide on epileptic rats induced by pentylenetetrazol. AB - OBJECTIVE: To investigate the effects of antisense glutamic acid decarboxylase (GAD(67)) oligodeoxynucleo-tide (ODN) on behavior, seizure threshold and EEG of hippocampus in the epileptic rats induced by pentylenetetrazol (PTZ). METHODS: A model of chronic epilepsy in rats was established by PTZ. The inhibition of GAD(67) mRNA expression in hippocampus was selectively induced by antisense oligodeoxynucleotide of GAD(67). The effect of antisense GAD(67) ODN on behavior, seizure threshold and EEG recording of kindled rats was examined. RESULTS: Antisense GAD(67) ODN could inhibit the expression of GAD(67) mRNA and the concentration of GABA. It also could significantly shorten the latencies of seizure and increase the level of seizure and the frequency of epileptiform discharges. CONCLUSION: The gene of GAD(67) may be an anti-seizure gene, which might inhibit epileptiform discharge. The treatment of epilepsy by GAD(67) gene will have a bright future. PMID- 11940382 TI - Genetic instability in cancer tissues analyzed by random amplified polymorphic DNA PCR. AB - OBJECTIVE: To detect DNA and chromosomes instabilities during the progression of tumors and screen new molecular markers coupled to putative or unknown oncogenes and/or tumor suppressor genes. METHODS: Five kinds of tumors, in a total of 128 specimens, were analyzed by random amplified polymorphic DNA (RAPD) PCR. Bands representing instabilities were recovered, purified, and cloned, labeled as probes for Southern and Northern blot analysis and DNA sequencing. RESULTS: Sample 5 and 3 of the gastric cancer tissues showed the highest genomic changes and the average detectability in five cancers was up to at least 40% (42.2% - 49.4%). There were significant differences in the ability of each primer to detect genomic instability, which ranged from 27% (primer 8) to 68% (primer 2). Band B is a single copy fragment, and was found to be an allelic loss in gastric and colon cancers. It is a novel sequence and was registered in GenBank with Accession Number AF151005. Further analysis revealed that it might be part of a cis-regulatory element of a new tumor suppressor gene, containing a promoter of cis-action "CACA" box, an enhancer of "GATA" family and a start codon. CONCLUSIONS: It was impossible or difficult to get great achievements for cancer treatments with the procedure of gene therapy only to one oncogene or one tumor suppressor gene because the extensive DNA variations occurred during the progression of tumor. RAPD assay connected with other techniques was a good tool for the detection of genomic instabilities and direct screening of some new molecular markers related to tumor suppressor genes or oncogenes. PMID- 11940383 TI - Effect of NF-kappaB on the induction of PDGF-B transcription by angiotensin II in the ECV304 cell line. AB - OBJECTIVE: To examine the effect of angiotensin II (Ang II) on nuclear factor kappa B (NF-kappaB) activation in human endothelial cell line ECV304 and the molecular mechanism by which Ang II activates NF-kappaB. METHODS: ECV304 cells were transiently cotransfected with an NF-kappaB/luciferase reporter gene and inactive NF-kappaB-inducing kinase (NIK), IkappaB kinase alpha (IKK(alpha)), IkappaB kinase beta (IKK(beta)) mutants or vectors, respectively. The effect on NF-kappaB was detected by using an electrophoretic mobility shift assay (EMSA) and overexpression of the mutants enabled blocking of reporter gene activation induced by Ang II. With immunofluorescence and immuno-electronic microscope techniques, including confocal microscopy and gold particle labeled electronic microscopy, definite cytoplasmic-to-nuclear translocations of NF-kappaB activation were detected using subunits p50 and p65 induced by Ang II. RESULTS: The translocation of p50 in nuclei was highly remarkable 2 hours after Ang II stimulation, and the activity was somewhat reduced 6 hours after stimulation to the 18th hour. Northern blot also showed PDGF-B mRNA increased by stimulation of Ang II for 18 hours. CONCLUSION: Ang II is effective in stimulating NF-kappaB activation through a pathway dependent on NIK, IKK(alpha) and IKK(beta), and induces PDGF-B transcription in the endothelial cell line, ECV304.v PMID- 11940384 TI - Multichannel piezoelectric genesensor for the detection of human papilloma virus. AB - OBJECTIVE: To establish a method for rapid detection and sub-typing of human papilloma virus (HPV). METHODS: We utilized the piezoelectric genosensor (PG) technique, which is a combination of the piezoelectric biosensor and gene chips for HPV identification in 22 recurrent biopsy specimens and 22 corresponding original biopsy specimens. The control samples came from normal tissue of healthy persons. A combined reaction took place on the sensor surface between the target genes and probes. The frequency of the piezoelectric sensor will decrease when such reactions occur, and the frequency decrease depends on the concentration of the target gene. Specimens were also analyzed with conventional PCR and dot blot. RESULTS: Of the 22 recurrent specimens, 15 contained HPV6 DNA, 2 HPV11 DNA, and 4 HPV16 DNA. Only one specimen was negative. All the 22 original specimens were positive: 17 harbored HPV6 DNA, 3 sequence homologous HPV11 DNA, and 2 HPV16 DNA. No HPV18 DNA was detected in any specimen. When compared with PCR and dot blot analysis, the results were essentially the same except for one specimen, which was shown to contain other sub-types of HPV. CONCLUSION: Our results show that the piezoelectric genosensor technique is a rapid and specific method to analyze HPV. PMID- 11940386 TI - Review of current progress in the structure and function of Smad proteins. AB - PURPOSE: To review the recent developments in the structure and function of Smad proteins. DATA SOURCES: Both Chinese- and English-language literatures were searched using MEDLINE/CD-ROM (1997 - 2000) and the Index of Chinese-Language Literature (1997 - 2000). STUDY SELECTION: Data from published articles about TGF beta signal transduction in recent domestic and foreign literature were selected. DATA EXTRACTION: Data were mainly extracted from 22 articles which are listed in the reference section of this review. RESULTS: Smad proteins mediate signal transduction induced by the TGF-beta superfamily. Based on their structural and functional properties, Smad proteins are divided into three groups. The first group, receptor-regulated Smads (R-Smads), are phosphorylated by activated type I receptors and form heteromeric complexes with the second group of Smads, common mediator Smads (Co-Smads). These Smad complexes translocate into the nucleus to influence gene transcription. Inhibitory Smads (I-Smads) are the third group and these antagonize the activity of R-Smads. In the nucleus, Smads can directly contact Smad-binding elements (SBE) in target gene promoters. Through interaction with different transcription factors, transcriptional co-activators or co repressors, Smads elicit different effects in various cell types. The aberrance of Smad proteins has been noted in several human disorders such as fibrosis, hypertrophic scarring and cancer. CONCLUSION: The structure of Smads determines their function as transcriptional factors which translocate signals from the cell surface to the nucleus where Smads regulate TGF-beta superfamily-dependent gene expression. PMID- 11940385 TI - Anti-endotoxic shock effects of cyproheptadine in rats. AB - OBJECTIVE: To investigate the antagonistic effect and mechanism of the effect of cyproheptadine (Cyp) on endotoxic shock in rats. METHODS: Endotoxic shock was produced in rats by i.v. injection of lipopolysaccharides (LPS) (5 mg/kg). Tumor necrosis factor (TNF(alpha)) mRNA expression was assessed by Northern blot. Plasma TNF(alpha) content was measured by radioimmunoassay. Plasma superoxide dismutase (SOD) activity and malondialdehyde (MDA) content were measured. The intracellular free calcium concentration ([Ca(2+)](i)) in single endothelial cells was determined by laser scanning confocal microscopy (LSCM). RESULTS: Cyp 5 mg/kg injected immediately after i.v. LPS raised the mean arterial blood pressure (MABP) of shocked rats and improved their 24 h survival rate. Meanwhile, Cyp markedly decreased TNF(alpha) mRNA levels in rat liver (18 +/- 10 vs. LPS + saline 38 +/- 10, P < 0.01) as well as plasma TNF(alpha) content [(7.8 +/- 2.4) microg/L vs. LPS + saline (21.5 +/- 3.2) microg/L, P < 0.01)]. It enhanced plasma SOD activity [(1037.2 +/- 112.8) NU/L vs LPS + saline (615.4 +/- 92.6) NU/L, P < 0.01], reduced the MDA content [(5.2 +/- 1.1) micromol/L vs. LPS + saline (9.8 +/ 1.5) micromol/L, P < 0.01], and inhibited TNF(alpha)-induced [Ca(2+)](i) elevation. CONCLUSION: Cyp exerts an anti-endotoxic shock effect by inhibiting TNF(alpha) gene expression, enhancing SOD activity, reducing lipid peroxidation, and preventing [Ca(2+)](i) overload. PMID- 11940388 TI - Development of a rat C6 brain tumor model. AB - OBJECTIVE: To establish rat C6 brain-tumor models and find a simple reliable index to judge tumor volume. METHODS: C6 cell suspension (10 microl) containing 10 g/L agarose and 1 x 10(6) cells was injected into the right caudate nucleus of the rat brain by a stereotaxic method. After implantation, rats were observed and given MRI scans. Rats were perfused with paraformaldehyde trans-aorta at the 10th, 15th, 20th and 25th day and before natural death. All brains, lungs, spinal cords and tumors were sectioned and inspected. Tumor-containing samples were prepared histologically by hematoxylin and eosin (HE) stains. RESULTS: Fifty implanted rats had 100% yield of intracerebral growth, with distant metastasis of 0% to 4%. CONCLUSION: A rat C6 brain-tumor model was successfully established. Rat survival time is correlated with tumor volume and may be useful as an index of tumor size. PMID- 11940387 TI - Epidermal growth factor concentrations in human milk, cow's milk and cow's milk based infant formulas. AB - OBJECTIVE: Because maternal epidermal growth factor (EGF) may be an adaptive response to accelerate growth and maturation in premature infants, we compared the EGF content in fresh cow's milk and cow's milk-based infant formulas with full and preterm mother's milk. METHODS: EGF content of 57 human colostrum from mothers delivering prematurely and at term, 4 different fresh cow's milk and 8 different cow's milk-based infant formulas was determined by radioimmunoassay (RIA). RESULTS: Human milk from mothers of premature infants had a higher EGF content compared to that from mothers of term infants (28.2 +/- 10.3 nmol/L vs. 17.3 +/- 9.6 nmol/L). EGF content in human milk negatively correlated with gestational age and birth weight of neonates. EGF content in fresh cow's milk (13.8 - 18.2 nmol/L) was similar to that in human term milk. EGF levels in non hydrolyzed protein formulas were much lower (5.6 - 8.6 nmol/L), and were undetectable in hydrolyzed protein formulas. CONCLUSION: The high EGF content in premature milk may represent a maternal compensatory mechanism to accelerate the growth and development of immature infants. Feeding infants with breast milk from their own mother should be advocated since there is lack of EGF in cow's milk based infant formulas. PMID- 11940389 TI - Brugada syndrome masquerading as acute myocardial infarction in a patient presenting with ventricular fibrillation. PMID- 11940390 TI - Amplatzer PDA occluder used in adult patent ductus arteriosus with serious pulmonary hypertension after testing balloon occlusion. PMID- 11940391 TI - [Effect of selective digestive decontamination on the nosocomial infection and multiresistant microorganisms incidence in critically ill patients]. AB - BACKGROUND: Ventilator-associated pneumonia (VAP) is a common complication in mechanically ventilated patients. Other infections related to the use of invasive devices, such urinary tract infections (UTI) and central venous catheter-related bloodstream infections are found in patients admitted in intensive care units (ICU). Selective digestive decontamination (SDD) is a set of manoeuvres (hygiene, topic digestive antimicrobials and short course of systemic antibiotics) aimed at decreasing the incidence of nosocomial infections, mainly VAP, in ICU. The objective of this study was to describe the impact of SDD in the rates of nosocomial infections in an 18-bed medical-surgical ICU. PATIENTS AND METHOD: All patients admitted in the ICU from October 1998 to September 1999 with expected mechanical ventilation for at least 72 h or with an illness where SDD has proven to be efficacious. The infection rates during this period were compared with the infection rates of the period without SDD (October 1997 to September 1998). RESULTS: VAP rates decreased from 12.38 to 3.64 per 1000 days of mechanical ventilation (RR 0.3; CI 95%: 0.16 to 0.53). Urinary tract infection rates decreased form 7.70 to 4.51 per 1000 bladder-catheter days (RR 0.6. CI 95%: 0.37 to 0.93). Central venous catheter related bloodstream infections decreased from 5.92 to 2.73 per 1000 catheter days (RR: 0.5. CI 95%: 0.24 to 0.90). There was no emergence of resistant microorganims when SDD was used. CONCLUSIONS: SDD reduces infection rates in ICU without any significant adverse events. PMID- 11940392 TI - [Prevention of vertical transmission of HIV-1 in Mallorca, Spain. Impact of antiretroviral therapy from 1995 to 2000]. AB - BACKGROUND: Our objective was to evaluate the impact of antiretroviral therapy (ART) in the prevention of maternal-fetal HIV transmission in a population of HIV infected pregnant women. PATIENTS AND METHOD: We studied prospectively all HIV infected pregnant women attended in our hospital from January 1995 to December 2000. We offered treatment with zidovudine (ZDV) alone or in combination according to women's requirements. RESULTS: There were 98 mother-infant pairs and we studied 93 of them. The rate of vertical transmission was 1.4% when ART was started in pregnancy. Risk of HIV transmission was greater in mothers not being treated with ART during pregnancy (relative risk [RR]: 18; 95% confidence interval [CI]: 2.2-145.4), in mothers who only received ZDV at delivery and child vs those who received ART during pregnancy (RR: 16.4; 95% CI: 1.8-145.6) and in mothers who were active intravenous drug users (RR: 9.3; 95% CI: 2.2-38.5), with significant differences between vaginal delivery and caesarean section. CONCLUSIONS: We observed a substantial benefit from ART, especially in the group of HIV-infected pregnant women who started treatment during pregnancy. Preventive interventions are needed. PMID- 11940393 TI - [Therapeutic compliance in patients with cardiovascular diseases]. AB - BACKGROUND: Our purpose was to establish, by means of a survival analysis, the duration of therapeutic compliance and the probability of abandonment or prescription drugs in cardiovascular patients, as well as the prognostic factors that determine it. PATIENTS AND METHOD: Longitudinal observational study (1996 1998). By means of a consecutive sampling, 493 patients who initiated a cardiovascular treatment were selected. Through interviews, we obtained information on cardiovascular problems and treatment, concomitant diseases, consumption of other drugs and social and demographic variables. The consumption of prescribed drugs was established across 6 periodic observations. RESULTS: During the observational period, 39.4% of drugs prescribed by the general practitioner (GP) were abandoned, as compared to 22.4% of those prescribed by specialists (p < 0.05). The degree of abandonment was significantly higher among consumers of vasodilators and vasoprotective agents. Cardiac glycosydes and angiotensin converting enzyme inhibitors were among the therapeutic subgroups in which a longer survival time was observed (average: 19.8 and 16.5 months, respectively). By a Cox regression analysis, we noticed that the risk of abandonment was higher in patients who took two or more doses of the drug per day (OR = 2.8; 95% IC, 21-37), in consumers of medicines with a daily cost lower than ptas. 100 (OR = 1.4); 95% CI, 1.0-1.8) and in subjects younger than 65 years (OR = 1.3; 95% CI, 1.0-1.8). CONCLUSIONS: A higher degree of abandonment of cardiovascular medication occurs when it is administered in primary health-care (i.e., drugs prescribed by the GP), mainly in relation to a greater prescription of agents with a low therapeutic effectiveness. Abandonment is influenced by patients' social and demographic factors and also by the specific characteristics of the treatment. PMID- 11940395 TI - [Adherence to antiretroviral therapy]. PMID- 11940394 TI - [Imported tuberculosis: an emerging disease in industrialised countries]. AB - BACKGROUND: The aim of this study was to describe the characteristics of imported tuberculosis (TB) in Barcelona during 1999 and 2000. MATERIAL AND METHOD: Epidemiological surveillance questionnaire. RESULTS: During 1999 and 2000, a 7.9% decrease in TB cases was observed among the native population in Barcelona, whereas cases among immigrants grew up to 47.2%. In 2000, 449 TB cases were detected among the native population (incidence, 29.5/100,000) and 121 among immigrants (incidence, 555.9/100,000). Three outbreaks were identified, involving one Indian community (11 cases), two Dominican families (4 cases) and one city school (2 cases) whose index case was a cooperant. Isolated strains of Mycobacterium tuberculosis at the first, second and third outbreak were multisensitive, multidrug-resistant and isoniazid-resistant, respectively. CONCLUSIONS: The emergence of imported TB cases in Barcelona over 1999 and 2000 suggests that current preventive guidelines must be reviewed. PMID- 11940396 TI - [Confidence intervals: something more than a statistical significance test]. PMID- 11940397 TI - [European Resuscitation Council Guidelines 2000 for adult basic life support]. PMID- 11940398 TI - [Abdominal pain and weight-loss in a 18-year-old male]. PMID- 11940399 TI - [Gas gangrene of the thigh secondary to Crohns disease of the sigmoid colon]. PMID- 11940400 TI - [Multiple sclerosis and ulcerative colitis: 'incidental concurrence or related diseases?]. PMID- 11940401 TI - [Community-acquired pneumonia in patients older than 60 years]. PMID- 11940403 TI - [Morganella morgagnii septic arthritis]. PMID- 11940404 TI - [Units of multiple sclerosis in Spanish public hospitals: are they feasible and necessary?]. PMID- 11940405 TI - [Descriptive study of first visits in a multiple sclerosis specialised unit]. AB - BACKGROUND: The need for optimal care has led to implementation of multiple sclerosis care units, specialised in diagnosis and follow-up of multiple sclerosis patients. Since information on multiple sclerosis prognosis is crucial, accurate diagnosis is one of their major roles. METHOD: Prospective analysis of first visits during a year (origin and diagnosis) at the Unidad de Neuroinmunologia Clinica of Hospital Vall d'Hebron. RESULTS: A total of 437 first visits were analysed; a moderate agreement (kappa = 0.468) between referral diagnoses and diagnosis at follow-up was obtained; agreement was very good (kappa = 0.844) between diagnosis at first visit and diagnosis at follow-up. In the subgroup of 200 patients with a demyelinating disease on follow-up after one year, 37.6% of referrals came from other non-specialised centers and 40.6% from our own center. CONCLUSION: Specialised care units enhance diagnostic precision; direct influence zone and non-specialised centers referrals are their more important sources of patients. PMID- 11940406 TI - [Discrepancy between imaging and neurophysiology in deep brain stimulation of medial pallidum and subthalamic nucleus in Parkinson's disease]. AB - OBJECTIVE: The objective of this work is to assess the discrepancy in distance between the target chosen by magnetic resonance imaging (MRI) and the final electrode placement after intraoperative microrecording in patients submitted to deep brain stimulation (DBS) for alleviating the Parkinson's disease (PD). METHODS: Thirty patients with PD and motor complications were operated with stereotactic surgery by MRI and microrecording. In 19 patients, the target chosen was the subthalamic nucleus (STN) and in 11 others the target was globus pallidus internus (GPi). In this work it is considered that the electrode has a current field below usual parameters of 1.5 mm radius. Consequently, when the distance error between the final physiological target and the MRI target, is between 1.5 and 3 mm was considered as partial discrepancy and distances of 3 mm or more were considered as total discrepancy. RESULTS: Partial discrepancy for STN and GPi were in 25 and 33% of the cases respectively and total discrepancy was 57 and 42% for each nucleus. The average distance error between both targets, final and image, for X stereotactic coordinate (mediolateral distance) was 1.54 mm for STN and 0.8 mm for GPi. The average distance for Y coordinate (anteroposterior distance) was 2.3 mm for STN and 2.2 mm for GPi. CONCLUSION: There is a significant discrepancy between the final physiological target after microrecording and the target chosen by MRI during surgery for alleviating PD that may induce variations or absence of clinical efficacy in parkinsonian patients submitted to the DBS surgery. Authors suggest the necessity of the microelectrode recording in order to reach the surgical target with the best clinical condition. PMID- 11940407 TI - Adaptation and standardization of the geriatric evaluation of relative's rating instrument (GERRI) for Spain. AB - BACKGROUND: The Geriatric Evaluation of Relative's Rating Instrument (GERRI) is a scale that evaluates the frequencies of alterations in behavior and functional capacity over a two-week period prior to exploration. The scale depends on the observations done by a relative o first caregiver of the studied subject. AIM: To adapt and standardize the GERRI for the use in the Spanish population as a part of a general project to standardized cognitive and functional tests. METHOD: The scale was administered to 444 subjects: 249 controls, 85 mild memory-cognitive disorders without dementia subjects (DWD) and 110 patients with Alzheimer-type dementia (ATD). An across-sectional statistical study was conducted in our samples stratified by age, gender and education. We evaluated the reliability of repeatability of the test, the internal reliability and the age, sex and education effects on the score of the different subscales. We also took into account the diagnostical validity in the Alzheimer disease and finally we correlated this test results with Mini mental test. RESULTS: The demographic variables age and schooling were found to affect the GERRI subscales differently. Gender did not reach significance. Internal consistency for the GERRI-Social, Mood and -Cognitive were 0.8620, 0.7647 and 0.9259, respectively. CONCLUSION: The Spanish version of the GERRI may be applied to Spanish clinical series because of its reliable internal consistency and reproducibility. PMID- 11940408 TI - [Immunophilins: neuroprotective agents and promoters of neural regeneration]. AB - Immunophilins are a family of proteins mainly known because they act as receptors of the immunosuppressant drugs cyclosporin A (CsA) and FK506. Immunophilins serve several general functions, including regulation of mitochondrial permeability, modulation of ion channels stability and acting as chaperones for a variety of proteins. However, immunophilins are also present at high density in the nervous system. CsA, FK506 and other derivatives inhibit the function of immunophilins and, through bloking or activating several intracellular pathways, it has been shown that they exert neuroprotective effects in different experimental models of ischemia, Parkinson's disease and excitotoxic insults. Moreover, FK506 also has neuroregenerative effects, by enhancing the axonal regeneration rate after lesions of the peripheral nervous system. The development of new agents that selectively bind to immunophilins opens new interesting perspectives for the therapy of degenerative diseases and injuries of the nervous system. PMID- 11940409 TI - [Short history of L-Dopa]. AB - The introduction of L-Dopa has been a landmark in Neuroscience. Before the use of L-Dopa, Parkinson's disease (PD) was considered a disabling disease with no effective treatment. The development of L-Dopa followed a rationale approach: first, the discovery of dopamine deficiency in the striatum of patients with PD; and then the use of precursor of dopamine. Overall, the whole process took less than 12 years; this is a remarkable short time taking into account that the development of a similar crucial drug such as insulin took 50 years. In addition, the success of L-Dopa stimulated the neurochemistry research for all neurodegenerative diseases. PMID- 11940410 TI - [Wallerian degeneration of the pyramidal bundle and amyotrophic lateral sclerosis in MRI]. PMID- 11940411 TI - [Prolonged clinical pattern of brain death in patients under barbiturate sedation: usefulness of transcranial Doppler]. AB - INTRODUCTION: Throughout the world, is fully accepted that a person is dead when brain death exists. In most situations, neurological criteria permit the diagnosis of brain death, but in some instances, as when high-dose barbiturate therapy has been used, confirmatory testing are required by law. CLINICAL CASE: We report the case of a 17 year-old women who suffered high-dose barbiturate therapy due to post traumatic intracranial hypertension. During the period of the barbiturate infusion and until six days after the suppression of this therapy, neurological exploration and EEG findings seem to confirm brain death, while transcranial Doppler (TCD) study remained normal. CONCLUSIONS: TCD is a fast, simple and accurate confirmatory testing in the determination of brain death and its findings are not affected by high-dose barbiturate therapy. We think that TCD must be present in all hospitals where mechanical ventilation and support of patients are carried out. PMID- 11940412 TI - Asymmetric myoclonic parietal syndrome in a patient with Alzheimer's disease mimicking corticobasal degeneration. AB - We describe a patient who presented a progressive asymmetrical parietal syndrome including ideomotor apraxia, hemiinattention, unilateral limb dystonia and myoclonus. The clinical picture of this patient supported the clinical diagnosis of corticobasal degeneration (CBD). However, the neuropathologic examination revealed abundant cortical betaA4-amyloid deposits, and phosphorylated tau accumulation in neuritic plaques, neurofibrillary tangles and neuropil threads corresponding to Alzheimer's disease (AD) stage V of Braak and Braak. This case supports the clinical heterogeneity in AD and the existence of a clinical overlap between AD and CBD. PMID- 11940413 TI - [Postsurgical sacral pseudomeningocele]. PMID- 11940414 TI - [Punding in Parkinsons disease]. PMID- 11940415 TI - [Pilot study with the monoclonal antibody IOR-C5 as a potential agent of radioimmunoscintigraphy in colorectal cancer]. AB - IOR C-5 is a G1 immunoglobulin type intact murine monoclonal antibody (MAb) that was developed in the Center of Molecular Immunology in Havana City, Cuba. In immunohistochemical studies, this demonstrated a significant affinity for the epithelial tissues so that it was used in a pilot clinical study to perform a radioimmunoscintigraphy of the colorectal primary tumors and their locoregional recurrences. It was labeled with 99mTc using the Schwarz method, with a > 95% performance. Planar images of the chest, abdomen and pelvis were performed at 10 minutes, 4-6 hours and 18-24 hours post-injection in the anterior and posterior projections and the SPECT was performed 4-6 hours and 18-24 hours post-injection of 1.85 GBq 99mTC. This study has aimed to verify in vivo the capacity of ior-C5 MAb to accumulate in the malignant colorectal lesions. ior-C5 accumulated in 5 out of the 7 patients who were studied and who were suffering from colorectal cancer or in whom there was suspicion of recurrence. There was a negative case of primary tumors, which was an adenocarcinoma in situ in a tubular-papillary adenoma. The second case with a negative radioimmunoscintigraphy was a true negative case. CONCLUSIONS: It can be concluded that even though the number of patients is quite low, ior-C5 fulfilled the expectations of recognizing the epitope expressed in colorectal tumors in an in vivo human environment. PMID- 11940416 TI - [Contribution of radioguided detection and selective biopsy of the sentinel lymph node to staging in 100 patients with cutaneous melanoma]. AB - OBJECTIVE: A selective sentinel node (SN) biopsy appears to be an alternative to conventional lymph node dissection for staging patients with cutaneous melanoma. This study has aimed to analyze our experience in the localization of the sentinel node with a probe detector and lymphoscintigraphy and its utility in the staging of this disease. MATERIAL AND METHODS: 100 patients, 56 female and 44 male, 51.5 31.5 mean age, diagnosed of cutaneous melanoma were studied. The lymphoscintigraphy was performed in all cases to detect the area of lymphatic drainage after peritumoral intradermal administration of 600 uCi of 99mTc colloidal sulfur, and planar images were acquired every 10 minutes until activity was detected in these areas. Afterwards, the intraoperative localization of the sentinel node with a probe detector was performed and the selective biopsy of the node was sent to the Pathology Department for its histologic study. RESULTS: The lymphoscintigraphy study was positive in 99 of the 100 cases and it was identified intraoperatively in 98 cases. The histologic analysis was negative in 78.9% and positive in 21.1% of the nodes. CONCLUSIONS: Radioisotopic lymphography and intraoperative probe detection are two very useful techniques for locating the sentinel node and staging patients with cutaneous melanoma. Many patients are benefiting from the selective biopsy of the sentinel node because of the decreased post-surgical morbidity and better staging of the cutaneous melanoma. PMID- 11940417 TI - [Breast cancer with metastatic invasion of the sentinel lymph node in an internal mammary chain and with axillary chains free of tumor lesionS]. AB - The lymphoscintigraphy is a widely accepted procedure to detect the sentinel node in breast cancer. This is a case report of a 45 year old woman diagnosed of breast cancer who was referred to the nuclear medicine facility to perform the sentinel node detection procedure. The lymphoscintigraphy prior to surgery shows a double pathway in the tracer migration: one going to the axillary area and the second to the internal mammary chain and shows hot spots that represent the sentinel nodes. After surgical removal of those nodes, the pathological analysis report indicated tumor infiltration in the internal mammary chain sentinel node with all axillary nodes free of malignant cells. The aim of this report is to emphasize the importance of performing a pathological analysis of all the sentinel nodes detected in lymphoscintigraphy. PMID- 11940419 TI - Nuclear cardiology in the new millennium. PMID- 11940418 TI - [Bone scintigraphy in two cases of primary vertebral osteosarcoma in adults]. AB - The primary vertebral osteosarcoma in adults is a rare tumor which represents less than 2% of all osteosarcomas. We present the cases of two men (40 and 33 years old) who began with pain and neurological compression symptoms. The imaging methods used to study the tumors were X-rays, CT, MRI and bone scintigraphy with 99mTc-HMDP. After the pathological diagnosis, the tumors were removed surgically and the treatment was completed with chemotherapy and radiotherapy. In this report the authors review the published cases of vertebral osteosarcoma, its epidemiology, clinical presentation and characteristics in the different imaging techniques. PMID- 11940420 TI - [Metastasis in leptomeninges discovered with PET-FDG]. PMID- 11940421 TI - Complete scintigraphic lesion regression after single 153Sm-EDTMP therapy in prostate cancer. PMID- 11940422 TI - [Evidence based medicine. Generalizations on the application to nuclear medicine. Part I]. PMID- 11940423 TI - [Cytosolic concentrations of cathepsin D in 88 infiltrating ductal breast carcinomas having negative estrogen and progesterone receptors. Correlation with other clinical and biological parameters]. PMID- 11940424 TI - [Physicians and justice (IV)]. PMID- 11940425 TI - [Comparative analysis of Light's criteria and other biochemical parameters to distinguish exudates from transudates]. AB - Light's criteria have classically been used to differentiate exudates from transudates. Nevertheless, a number of studies have attempted to identify more efficient parameters. The objective of our study was to determine the usefulness of biochemical parameters to differentiate transudates from exudates, and to compare them with the so far best studied criteria: the Light's criteria. We prospectively analysed 850 non selected cases of pleural effusion, with closed final diagnosis after its confirmation, therapeutic response and follow-up, collected consecutively at the Pleura Unit of our hospital. The parameters evaluated as potentially discriminatory between transudates and exudates included: glucose, proteins, albumin, lactate-dehydrogenase (LDH), cholesterol, triglycerides, bilirubin, alkaline phosphatase and adenosin-deaminase (ADA), both separately and in combination to obtain the highest yield. The highest diagnostic yield was observed with the combination of pleural cholesterol, pleural LDH, and the pleural fluid/serum protein ratio, but without significant differences between combinations of pleural cholesterol and LDH, pleaural LDH and pleural proteins, Light's criteria or modified Light's criteria. We recommend the use of pleural cholesterol higher than 47 mg/dl and pleural LDH higher than 222 IU/l to offer the same yield as the combination of three parameters, due to its lower cost and because the necessity of serum determinations is avoided. PMID- 11940426 TI - [Binswanger's disease or multi-infarct dementia? Diagnostic keys in vascular dementia]. AB - OBJECTIVE: To develop a model that assists in the classification of patients with vascular dementia in two of its most relevant subtypes: multi-infarct dementia (cortical involvement) and Binswanger's disease (subcortical involvement). PATIENTS AND METHODS: Retrospective analysis of 50 consecutive patients with Binswanger's disease and 50 patients with multi-infarct dementia studied throughout a 12-year period (1986-1998) in a tertiary university hospital. The statistical model was obtained by multivariant analysis (logit model) and its accuracy was estimated by means of ROC curve analysis. RESULTS: The variables included in the regression model were: arterial hypertension (p < 0.03), cardioembolic disease (p < 0.05), and presence of cortical (p < 0.01) and lacunar (p < 0.01) infarctions in CAT. The specificity of the model was 90%, the sensitivity near 60%, and the positive likelihood ratio 5.8. The area under the ROC curve was 0.80 (0.73-0.87). CONCLUSIONS: Despite the availability of consensus diagnostic criteria for vascular dementia, their use is very commonly restricted to the research field. We have developed a classification model for patients with vascular dementia in two relevant subgroups: multi-infarct dementia and Binswanger's disease. This model can contribute to make identification of these patients in the daily clinical practice easier. PMID- 11940427 TI - [The problem of arterial hypertension in Spain]. PMID- 11940428 TI - [The microbiology laboratory and tuberculosis]. PMID- 11940429 TI - [Diagnostic criteria and differential diagnosis of Behcets disease]. PMID- 11940430 TI - [Advances in the treatment of osteoporosis at the beginning of the new millennium]. PMID- 11940431 TI - [Lipodystrophy syndrome associated with the human immunodeficiency syndrome virus (I). General characteristics and etiopathogenesis]. PMID- 11940432 TI - [The physicians and justice (VI): should I hand the clinical record over or is a report enough?]. PMID- 11940433 TI - [Progressive anemia and early postpandrial fullness]. PMID- 11940434 TI - [Ileitis with recurrent ascites]. PMID- 11940435 TI - [Abdominal pain and secondary polyglobulia]. PMID- 11940436 TI - [Abdominal mass in a young patient]. PMID- 11940437 TI - [Night sweat without fever. Why not gastroesphageal reflux?]. PMID- 11940438 TI - [Hypophyseal resistance to thyroid hormones]. PMID- 11940439 TI - [Refractory idiopathic thrombocytopenic purpura and selective IgA deficiency]. PMID- 11940440 TI - [Arsenic poisoning: a diagnostic problem]. PMID- 11940442 TI - Safety of carrageenan in foods. PMID- 11940443 TI - Cancer. Plants provide prevention. PMID- 11940444 TI - The Hudson: a river runs through an environmental controversy. PMID- 11940445 TI - e-Junk explosion. PMID- 11940446 TI - Who pays for e-Junk? PMID- 11940447 TI - Leading the charge for better batteries. PMID- 11940448 TI - The association of blood lead level and cancer mortality among whites in the United States. AB - Lead is classified as a possible carcinogen in humans. We studied the relationship of blood lead level and all cancer mortality in the general population of the United States using data from the National Health and Nutrition Examination Survey II (NHANES II) Mortality Study, 1992, consisting of a total of 203 cancer deaths (117 men and 86 women) among 3,592 whites (1,702 men and 1,890 women) with average of 13.3 years of follow-up. We used Cox proportional hazard regression models to estimate the dose-response relationship between blood lead and all cancer mortality. Log-transformed blood lead was either categorized into quartiles or treated as a continuous variable in a cubic regression spline. Relative risks (RRs) were estimated for site-specific cancers by categorizing lead above and below the median. Among men and women combined, dose-response relationship between quartile of blood lead and all cancer mortality was not significant (ptrend = 0.16), with RRs of 1.24 [95% percent confidence interval (CI), 0.66-2.33], 1.33 (95% CI, 0.57-3.09), and 1.50 (95% CI, 0.75-3.01) for the second, third, and fourth quartiles, respectively, compared with the first quartile. Spline analyses found no dose response (p = 0.29), and none of the site specific cancer RRs were significant. Among men, no significant dose-response relationships were found for quartile or spline analyses (p trend = 0.57 and p = 0.38, respectively). Among women, no dose-response relationship was found for quartile analysis (ptrend = 0.22). However, the spline dose-response results were significant (p = 0.001), showing a threshold effect at the 94th percentile of blood lead or a lead concentration of 24 microg/dL, with an RR of 2.4 (95% CI, 1.1-5.2) compared with the risk at 12.5 percentile. Because the dose-response relationship found in women was not found in men, occurred at only the highest levels of lead, and has no clear biologic explanation, further replication of this relationship is needed before it can be considered believable. In conclusion, individuals with blood lead levels in the range of NHANES II do not appear to have increased risk of cancer mortality. PMID- 11940449 TI - Evidence for induction of oxidative stress caused by chronic exposure of Chinese residents to arsenic contained in drinking water. AB - Exposure of experimental animals or cultured cells to arsenic induces oxidative stress, but, to date, no examination of this phenomenon in humans has been reported. In this study we conducted a cross-sectional study in Wuyuan, Inner Mongolia, China, to explore the relationship between chronic arsenic exposure from drinking water and oxidative stress in humans. Thirty-three inhabitants who had been drinking tube-well water with high concentrations of inorganic arsenic (mean value = 0.41 mg/L) for about 18 years constituted the high-exposure group, and 10 residents who lived nearby but were exposed to much lower concentrations of arsenic in their drinking water (mean value = 0.02 mg/L) were selected as the low-exposure comparison group. Results of the present study indicated that although the activity for superoxide dismutase (SOD) in blood did not differ significantly between the two groups, the mean serum level of lipid peroxides (LPO) was significantly higher among the high-exposed compared with the low exposed group. Elevated serum LPO concentrations were correlated with blood levels of inorganic arsenic and its methylated metabolites. In addition, they showed an inverse correlation with nonprotein sulfhydryl (NPSH) levels in whole blood. The subjects in the high-arsenic-exposure group had mean blood NPSH levels 57.6% lower than those in the low-exposure group. Blood NPSH levels were inversely correlated with the concentrations of inorganic arsenic and its methylated metabolites in blood and with the ratio of monomethylarsenic to inorganic arsenic. These results provide evidence that chronic exposure to arsenic from drinking water in humans results in induction of oxidative stress, as indicated by the reduction in NPSH and the increase in LPO. Some possible mechanisms for the arsenic-induced oxidative stress are discussed. PMID- 11940450 TI - Renal effects of uranium in drinking water. AB - Animal studies and small studies in humans have shown that uranium is nephrotoxic. However, more information about its renal effects in humans following chronic exposure through drinking water is required. We measured uranium concentrations in drinking water and urine in 325 persons who had used drilled wells for drinking water. We measured urine and serum concentrations of calcium, phosphate, glucose, albumin, creatinine, and beta-2-microglobulin to evaluate possible renal effects. The median uranium concentration in drinking water was 28 microg/L (interquartile range 6-135, max. 1,920 microg/L) and in urine 13 ng/mmol creatinine (2-75), resulting in the median daily uranium intake of 39 microg (7-224). Uranium concentration in urine was statistically significantly associated with increased fractional excretion of calcium and phosphate. Increase of uranium in urine by 1 microg/mmol creatinine increased fractional excretion of calcium by 1.5% [95% confidence interval (CI), 0.6-2.3], phosphate by 13% (1.4-25), and glucose excretion by 0.7 micromol/min (-0.4-1.8). Uranium concentrations in drinking water and daily intake of uranium were statistically significantly associated with calcium fractional excretion, but not with phosphate or glucose excretion. Uranium exposure was not associated with creatinine clearance or urinary albumin, which reflect glomerular function. In conclusion, uranium exposure is weakly associated with altered proximal tubulus function without a clear threshold, which suggests that even low uranium concentrations in drinking water can cause nephrotoxic effects. Despite chronic intake of water with high uranium concentration, we observed no effect on glomerular function. The clinical and public health relevance of the findings are not easily established, but our results suggest that the safe concentration of uranium in drinking water may be within the range of the proposed guideline values of 2-30 microg/L. PMID- 11940451 TI - Hydroxylated polychlorinated biphenyls as inhibitors of the sulfation and glucuronidation of 3-hydroxy-benzo[a]pyrene. AB - Polychlorinated biphenyls (PCBs) can be metabolized by cytochromes P450 to hydroxylated biotransformation products. In mammalian studies, some of the hydroxylated products have been shown to be strong inhibitors of steroid sulfotransferases. As a part of ongoing research into the bioavailability of environmental pollutants in catfish intestine, we investigated the effects of a series of hydroxylated PCBs (OH-PCBs) on two conjugating enzymes, phenol-type sulfotransferase and glucuronosyltransferase. We incubated cytosolic and microsomal samples prepared from intestinal mucosa with 3-hydroxy-benzo[a]pyrene and appropriate cosubstrates and measured the effect of OH-PCBs on the formation of BaP-3-glucuronide and BaP-3-sulfate. We used PCBs with 4, 5, and 6 chlorine substitutions and the phenolic group in the ortho, meta, and para positions. OH PCBs with the phenolic group in the ortho position were weak inhibitors of sulfotransferase; the median inhibitory concentration (IC50) ranged from 330 to 526 microM. When the phenol group was in the meta or para position, the IC50 was much lower (17.8-44.3 microM). The OH-PCBs were more potent inhibitors of glucuronosyltransferase, with IC50s ranging from 1.2 to 36.4 microM. The position of the phenolic group was not related to the inhibitory potency: the two weakest inhibitors of sulfotransferase, with the phenolic group in the ortho position, were 100 times more potent as inhibitors of glucuronosyltransferase. Inhibition of glucuronosyltransferase by low concentrations of OH-PCBs has not been reported before and may have important consequences for the bioavailability, bioaccumulation, and toxicity of other phenolic environmental contaminants. PMID- 11940452 TI - Alteration of intracellular cysteine and glutathione levels in alveolar macrophages and lymphocytes by diesel exhaust particle exposure. AB - The purpose of this study was to characterize the effects of diesel exhaust particles (DEP) on thiol regulation in alveolar macrophages (AM) and lymphocytes. We obtained AM and lymph node (thymic and tracheal) cells (LNC) (at different time points) from rats exposed intratracheally to DEP (5 mg/kg) or saline, and measured inflammatory markers, thiol levels, and glutathione reductase (GSH-R) activity. DEP exposure produced significant increases in neutrophils, lactate dehydrogenase, total protein, and albumin content in the lavage fluid. AM from DEP-exposed rats showed a time-dependent increase in intracellular cysteine (CYSH) and GSH. In LNC the intracellular GSH reached peak level by 24 hr, declining toward control levels by 72 hr after exposure. LNC-CYSH and AM-CYSH and GSH were increased at both 24 and 72 hr. Both Sprague-Dawley and Brown Norway rats showed similar trends of responses to DEP exposure as per measurement of the inflammatory markers and thiol changes. AM and, to a lesser degree, LNC were both active in cystine uptake. The DEP exposure stimulated GSH-R activity and increased the conversion of cystine to CYSH in both cell types. The intracellular level of GSH in DEP-exposed AM was moderately increased compared with the saline control, and was further augmented when cells were incubated with cystine. In contrast, the intracellular level of GSH in DEP-exposed LNC was significantly reduced despite the increased CYSH level and GSH-R activity when these cells were cultured for 16 hr. DEP absorbed 23-31% of CYSH, cystine, and GSH, and only 8% of glutathione disulfide when incubated in cell free media. These results indicate that DEP exposure caused lung inflammation and affected thiol levels in both AM and LNC. PMID- 11940453 TI - Polychlorinated dibenzo-p-dioxins, dibenzofurans, and biphenyls in fishermen in Finland. AB - We measured plasma concentrations of polychlorinated dibenzo-p-dioxins and dibenzofurans (PCDD/Fs), and polychlorinated biphenyls (PCBs) in fishermen from the Finnish Baltic Sea area and fishermen fishing in inland lakes. The concentrations clearly correlated with the frequency of fish meals and consumption of Baltic fatty fish. The body burden of PCDD/Fs reached the median level of 170 pg/g toxic equivalents (I-TEq) in fat for Baltic Sea fishermen, with the maximum being 420 pg/g. Results for 2,3,7,8-tetrachlorodibenzo-p-dioxin (range = 4.9-110 pg/g fat) showed that lifetime exposure in a population consuming much Baltic fatty fish can reach the levels of exposures seen in Seveso, Italy, in 1976. After we summed the PCB-TEqs, the total median exposure of Baltic Sea fishermen increased to 290 pg/g TEq in fat, and the highest concentration was 880 pg/g. There was a noted individual variation in fishermen's PCDD/F congener patterns, and it was possible to associate this variation with congener patterns of PCDD/Fs in the fish species that the fisherman reported they had consumed. Linear regression models for ln WHO(PCDD/F)-TEq, ln WHO(PCB)-TEq, and ln total WHO-TEq, from the World Health Organization, explained 48%, 60%, and 53% of the variability, respectively. Age was the only significant predictor of ln WHO(PCDD/F)-TEq, whereas age, amount of fish eaten, and place of residence were significant predictors of ln WHO(PCB)-TEq, and ln total WHO-TEq. PMID- 11940454 TI - Prediction of rodent nongenotoxic carcinogenesis: evaluation of biochemical and tissue changes in rodents following exposure to nine nongenotoxic NTP carcinogens. AB - We studied nine presumed nongenotoxic rodent carcinogens, as defined by the U.S. National Toxicology Program (NTP), to determine their ability to induce acute or subacute biochemical and tissue changes that may act as useful predictors of nongenotoxic rodent carcinogenesis. The chemicals selected included six liver carcinogens (two of which are peroxisome proliferators), three thyroid gland carcinogens, and four kidney carcinogens. We administered the chemicals (diethylhexyl phthalate, cinnamyl anthranilate, chlorendic acid, 1,4 dichlorobenzene, monuron, ethylene thiourea, diethyl thiourea, trimethyl thiourea, and d-limonene to the same strains of mice and rats used in the original NTP bioassays (nine chemicals to rats and seven to mice). Selected tissues (liver, thyroid gland, and kidney) were collected from groups of animals at 7, 28, and 90 days for evaluation. Tissue changes selected for study were monitored for all of the test groups, irrespective of the specificity of the carcinogenic responses observed in those tissues. This allowed us to assess both the carcinogen specificity and the carcinogen sensitivity of the events being monitored. We studied relative weight, cell labeling indices, and pathologic changes such as hypertrophy in all tissues; a range of cytochrome P450 enzymes and palmitoyl coenzyme A oxidase in the liver; changes in the levels of plasma total triiodothyronine, total thyroxine, and thyroid-stimulating hormone (TSH) as markers of thyroid gland function; and hyaline droplet formation, tubular basophilia, and the formation of granular casts in the kidney. There were no single measurements that alerted specifically to the carcinogenicity of the agents to the rodent liver, thyroid gland, or kidney. However, in the majority of cases, the chemical induction of cancer in a tissue was preceded by a range of biochemical/morphologic changes, most of which were moderately specific for a carcinogenic outcome, and some of which were highly specific for it (e.g., increases in TSH in the thyroid gland and increases in relative liver weight in the mouse). The only measurements that failed to correlate usefully with carcinogenicity were the induction of liver enzymes (with the exception of the enzymes associated with peroxisome proliferation). Most of the useful markers were evident at the early times studied (7 days and 28 days), but no overall best time for the measurement of all markers was identified. The judicious choice of markers and evaluation times can aid the detection of potential nongenotoxic rodent carcinogens. PMID- 11940456 TI - The potential impact of flooding on confined animal feeding operations in eastern North Carolina. AB - Thousands of confined animal feeding operations (CAFOs) have been constructed in eastern North Carolina. The fecal waste pit and spray field waste management systems used by these operations are susceptible to flooding in this low-lying region. To investigate the potential that flood events can lead to environmental dispersion of animal wastes containing numerous biologic and chemical hazards, we compared the geographic coordinates of 2,287 CAFOs permitted by the North Carolina Division of Water Quality (DWQ) with estimates of flooding derived from digital satellite images of eastern North Carolina taken approximately 1 week after Hurricane Floyd dropped as much as 15-20 inches of rain in September 1999. Three cattle, one poultry, and 237 swine operations had geographic coordinates within the satellite-based flooded area. DWQ confirmed 46 operations with breached or flooded fecal waste pits in the same area. Only 20 of these 46 CAFOs were within the satellite-based estimate of the inundated area. CAFOs within the satellite-based flood area were located in 132 census block groups with a population of 171,498 persons in the 2000 census. African Americans were more likely than whites to live in areas with flooded CAFOs according to satellite estimates, but not according to DWQ reports. These areas have high poverty rates and dependence on wells for drinking water. Our analysis suggests that flood events have a significant potential to degrade environmental health because of dispersion of wastes from industrial animal operations in areas with vulnerable populations. PMID- 11940455 TI - A new invertebrate member of the p53 gene family is developmentally expressed and responds to polychlorinated biphenyls. AB - The cell-cycle checkpoint protein p53 both directs terminal differentiation and protects embryos from DNA damage. To study invertebrate p53 during early development, we identified three differentially expressed p53 family members (p53, p97, p120) in the surf clam, Spisula solidissima. In these mollusks, p53 and p97 occur in both embryonic and adult tissue, whereas p120 is exclusively embryonic. We sequenced, cloned, and characterized p120 cDNA. The predicted protein, p120, resembles p53 across all evolutionarily conserved regions and contains a C-terminal extension with a sterile alpha motif (SAM) as in p63 and p73. These vertebrate forms of p53 are required for normal inflammatory, epithelial, and neuronal development. Unlike clam p53 and p97, p120 mRNA and protein levels are temporally expressed in embryos, with mRNA levels decreasing with increasing p120 protein (R(2) = 0.97). Highest surf clam p120 mRNA levels coincide with the onset of neuronal growth. In earlier work we have shown that neuronal development is altered by exposure to polychlorinated biphenyls (PCBs), a neurotoxic environmental contaminant. In this study we show that PCBs differentially affect expression of the three surf clam p53 family members. p120 mRNA and protein are reduced the most and earliest in development, p97 protein shows a smaller and later reduction, and p53 protein levels do not change. For the first time we report that unlike p53 and p97, p120 is specifically embryonic and expressed in a time-dependent manner. Furthermore, p120 responds to PCBs by 48 hr when PCB-induced suppression of the serotonergic nervous system occurs. PMID- 11940457 TI - A benchmark dose analysis of prenatal exposure to polychlorinated biphenyls. AB - Benchmark dose (BMD) analysis is used to determine levels of exposure to environmental contaminants associated with increased public health risk. In this study we used a benchmark approach to evaluate the risks associated with prenatal exposure to polychlorinated biphenyls (PCBs). We evaluated for intellectual impairment a cohort of children whose prenatal PCB exposure had been assessed from biologic specimens. We calculated BMDs and lower-bound confidence limits (BMDLs) for four end points using four sets of risk criteria. BMDLs were estimated using three different statistical methodologies. The BMDs and BMDLs were remarkably consistent across the four end points for each set of risk criteria, but differed substantially for the different risk criteria. The proportion of the sample considered at risk ranged from 9.8% for the least protective criteria to 74.1% for the most protective. Two methodologies, likelihood ratio and bootstrapping, generated generally similar BMDLs. BMD analysis provides a straightforward, reliable method for evaluating levels of exposure associated with increased public health risk. In the analyses performed in this study, the number of individuals considered at risk depended more on the risk criterion selected than on the outcome assessed. PMID- 11940458 TI - Sister chromatid exchanges and micronuclei in peripheral lymphocytes of shoe factory workers exposed to solvents. AB - We examined sister chromatid exchanges (SCEs) and micronuclei (MN; cytokinesis block method) in cultured peripheral lymphocytes from 52 female workers of two shoe factories and from 36 unexposed age- and sex-matched referents. The factory workers showed an elevated level of urinary hippuric acid, a biomarker of toluene exposure, and workplace air contained high concentrations of various organic solvents such as toluene, gasoline, acetone, and (in one of the plants only) ethylacetate and methylenediphenyl diisocyanate. The shoe factory workers showed a statistically significant higher frequency of micronucleated binucleate lymphocytes in comparison with the referents. This finding agreed with three preliminary MN determinations (each comprising 27-32 shoe workers and 16-20 controls) performed in one of the plants 2-5 years earlier. The shoe factory workers also had a lower average level of blood hemoglobin than the referents. In contrast, no difference was found between the groups in SCE analysis. Smokers showed significantly higher mean frequencies of SCEs per cell and high frequency cells (HFC) than nonsmokers. Aging was associated with increased MN rates and reduced cell proliferation. Polymorphism of the glutathione S-transferase M1 gene (GSTM1) did not affect the individual level of SCEs; but in smoking shoe workers an effect of the occupational exposure on the frequency of micronucleated cells could be seen only in GSTM1 null subjects. The low prevalence of the glutathione S-transferase T1 (GSTT1) null genotype precluded the evaluation of the influence of GSTT1 polymorphism. Our results show that the shoe factory workers have experienced genotoxic exposure, which is manifest as an increase in the frequency of MN, but not of SCEs, in peripheral lymphocytes. The exposures responsible for the MN induction could not be identified with certainty, but exposure to benzene in gasoline and methylenediphenyl diisocyanate may explain some of the findings. PMID- 11940459 TI - The association between biomarker-based exposure estimates for phthalates and demographic factors in a human reference population. AB - Population-based estimates of environmental exposures using biomarkers can be difficult to obtain for a variety of reasons, including problems with limits of detection, undersampling of key strata, time between exposure and sampling, variation across individuals, variation within individuals, and the ability to find and interpret a given biomarker. In this article, we apply statistical likelihoods, weighted sampling, and regression methods for censored data to the analysis of biomarker data. Urinary metabolites for seven phthalates, reported by Blount et al., are analyzed using these methods. In the case of the phthalates data, we assumed the underlying model to be a log-normal distribution with the mean of the distribution defined as a function of a number of demographic variables that might affect phthalate levels in individuals. Included as demographic variables were age, sex, ethnicity, residency, family income, and education level. We conducted two analyses: an unweighted analysis where phthalate distributions were estimated with changes in the means of these distributions as a function of demographic variables, and a weighted prediction for the general population in which weights were assigned for a subset of the population depending on the frequency of their demographic variables in the general U.S. population. We used statistical tests to determine whether any of the demographic variables affected mean phthalate levels. Individuals with only a high school education had higher levels of di-n-butyl phthalate than individuals with education beyond high school. Subjects who had family income less than $1,500 in the month before sampling and/or only high school education had higher levels of n-butyl benzyl phthalate levels than other groupings. Di(2-ethylhexyl) phthalate was higher in males and/or in urban populations and/or in people who had family income less than $1,500 per month. Our findings suggest that there may be significant demographic variations in exposure and/or metabolism of phthalates and that health-risk assessments for phthalate exposure in humans should consider different potential risk groups. PMID- 11940460 TI - Pentachlorophenol and hydroxylated polychlorinated biphenyl metabolites in umbilical cord plasma of neonates from coastal populations in Quebec. AB - Concentrations of polychlorinated biphenyls (PCBs), hydroxylated metabolites of PCBs (HO-PCBs) and octachlorostyrene (4-HO-HpCS), and pentachlorophenol (PCP) were determined in umbilical cord plasma samples from three different regions of Quebec. The regions studied included two coastal areas where exposure to PCBs is high because of marine-food-based diets--Nunavik (Inuit people) and the Lower North Shore of the Gulf of St. Lawrence (subsistence fishermen)--and a southern Quebec urban center where PCB exposure is at background levels (Quebec City). The main chlorinated phenolic compound in all regions was PCP. Concentrations of PCP were not significantly different among regions (geometric mean concentration 1,670 pg/g, range 628-7,680 pg/g wet weight in plasma). The ratio of PCP to polychlorinated biphenyl congener number 153 (CB153) concentration ranged from 0.72 to 42.3. Sum HO-PCB (sigma HO-PCBs) concentrations were different among regions, with geometric mean concentrations of 553 (range 238-1,750), 286 (103 788), and 234 (147-464) pg/g wet weight plasma for the Lower North Shore, Nunavik, and the southern Quebec groups, respectively. Lower North Shore samples also had the highest geometric mean concentration of sum PCBs (sum of 49 congeners; sigma PCBs), 2,710 (525-7,720) pg/g wet weight plasma. sigma PCB concentrations for Nunavik samples and southern samples were 1,510 (309-6,230) and 843 (290-1,650) pg/g wet weight plasma. Concentrations (log transformed) of sigma HO-PCBs and sigma PCBs were significantly correlated (r = 0.62, p < 0.001), as were concentrations of all major individual HO-PCB congeners and individual PCB congeners. In Nunavik and Lower North Shore samples, free thyroxine (T4) concentrations (log transformed) were negatively correlated with the sum of quantitated chlorinated phenolic compounds (sum PCP and sigma HO-PCBs; r = -0.47, p = 0.01, n = 20) and were not correlated with any PCB congeners or sigma PCBs. This suggests that PCP and HO-PCBs are possibly altering thyroid hormone status in newborns, which could lead to neurodevelopmental effects in infants. Further studies are needed to examine the effects of chlorinated phenolic compounds on thyroid hormone status in newborns. PMID- 11940462 TI - Summary of the National Toxicology Program's report of the endocrine disruptors low-dose peer review. AB - At the request of the U.S. Environmental Protection Agency (U.S. EPA), the National Toxicology Program organized an independent and open peer review to evaluate the scientific evidence on low-dose effects and nonmonotonic dose response relationships for endocrine-disrupting chemicals in mammalian species. For this peer review, "low-dose effects" referred to biologic changes that occur in the range of human exposures or at doses lower than those typically used in the standard testing paradigm of the U.S. EPA for evaluating reproductive and developmental toxicity. The demonstration that an effect is adverse was not required because in many cases the long-term health consequences of altered endocrine function during development have not been fully characterized. A unique aspect of this peer review was the willing submission of individual animal data by principal investigators of primary research groups active in this field and the independent statistical reanalyses of selected parameters prior to the peer review meeting by a subpanel of statisticians. The expert peer-review panel (the panel) also considered mechanistic data that might influence the plausibility of low-dose effects and identified study design issues or other biologic factors that might account for differences in reported outcomes among studies. The panel found that low-dose effects, as defined for this review, have been demonstrated in laboratory animals exposed to certain endocrine-active agents. In some cases where low-dose effects have been reported, the findings have not been replicated. The shape of the dose-response curves for reported effects varied with the end point and dosing regimen and were low-dose linear, threshold-appearing, or nonmonotonic. The findings of the panel indicate that the current testing paradigm used for assessments of reproductive and developmental toxicity should be revisited to see whether changes are needed regarding dose selection, animal model selection, age when animals are evaluated, and the end points being measured following exposure to endocrine-active agents. PMID- 11940463 TI - The pitfalls of hair analysis for toxicants in clinical practice: three case reports. AB - Hair analysis is used to assess exposure to heavy metals in patients presenting with nonspecific symptoms and is a commonly used procedure in patients referred to our clinic. We are frequently called on to evaluate patients who have health related concerns as a result of hair analysis. Three patients first presented to outside physicians with nonspecific, multisystemic symptoms. A panel of analytes was measured in hair, and one or more values were interpreted as elevated. As a result of the hair analysis and other unconventional diagnostic tests, the patients presented to us believing they suffered from metal toxicity. In this paper we review the clinical efficacy of this procedure within the context of a patient population with somatic disorders and no clear risk factors for metal intoxication. We also review limitations of hair analysis in this setting; these limitations include patient factors such as low pretest probability of disease and test factors such as the lack of validation of analytic techniques, the inability to discern between exogenous contaminants and endogenous toxicants in hair, the variability of analytic procedures, low interlaboratory reliability, and the increased likelihood of false positive test results in the measurement of panels of analytes. PMID- 11940461 TI - Dust mite, cockroach, cat, and dog allergen concentrations in homes of asthmatic children in the northeastern United States: impact of socioeconomic factors and population density. AB - Home exposures to aeroallergens are an important environmental factor in allergic sensitization and in the development and exacerbation of asthma. We assessed variations in home concentrations of dust mite, cockroach, cat, and dog allergens in dust collected in the main living areas of asthmatics' homes by family income, mother's education, dwelling type, population density, household population density, and ethnicity in Connecticut and south-central Massachusetts. Dust samples were collected at the time of home interview in 999 homes as part of an ongoing longitudinal birth cohort study of 1,002 infants and their asthmatic siblings. The analysis employed lower and upper cut points for group 1 dust mite (> or = 2.0 microg/g and > or = 10 microg/g), cockroach (> or = 1.0 U/g and > or = 4.0 U/g), cat (> or = 1.0 microg/g and > or = 8.0 ug/g), and dog (> or = 2.0 microg/g and > or = 10.0 microg/g) allergens. Subject residences were geocoded to assess population density from the U.S. Census, and multiple logistic regression was used to control for confounding. The portion of homes at the lower cut point for dust mite, cockroach, cat, and dog allergens were 46.9%, 24.9%, 42.2%, and 35.6%, respectively; the upper cut point for each of the allergens was reached in 22.4%, 13.4%, 21.0%, and 22.9% of the homes, respectively. In all, 86.0% of the homes had at least one allergen at the lower cut point, and 58.0% had at least one allergen at the upper cut point. Forty-nine percent of the homes had two or more allergens at the lower cut point, and 19.7% had two or more allergens at the upper cut point. Higher education of the mother, higher household income, living in a single-family home in a less densely populated area with fewer people per room, and being a white household were associated with elevated dust mite, cat, and dog allergens and low cockroach allergen. In contrast, low income, living in a multifamily home in a high population density area with a higher occupancy rate per room, and being a Hispanic or black household were associated with elevated cockroach allergens and low concentrations of dust mite, cat, and dog allergens. Although the presence of an individual allergen is more likely associated with one or more socioeconomic or ethnic factors, most homes typically have multiple allergen burdens in excess of concentrations thought to be associated with sensitization and exacerbation of asthma. Mite and cockroach allergens have distinct and opposite associations with socioeconomic factors and population density. PMID- 11940465 TI - [Protein kinase CK2 and cancer: further clues are accumulating]. AB - Protein kinase CK2 appears to interact with different signaling pathways and therefore represents the prototype of a multifunctional protein kinase. The enzyme is highly expressed in most cancers and this higher expression has been tentatively correlated with the involvement of CK2 in the promotion of specific phases of the cell cycle. This review summarizes our current knowledge on how CK2 may also contribute to tumorigenesis through its direct interaction with the cell survival circuitry. PMID- 11940464 TI - [The hypoxia-inducible factor HIF as a new target in cancer research]. AB - The vascular endothelial growth factors (VEGF) plays major roles during tumor development. VEGF expression is regulated by the transcription factor HIF (hypoxia inducible factor) which is very sensitive to oxygene variations. We will show here that HIF1alpha is today a good prognostic and diagnostic marker for an increasing number of cancers from various origins. Furthermore, the reduction of the expression or the activity of HIF1alpha impairs tumour angiogenesis and subsequently growth. Altogether, these results suggest that HIF1alpha could be an interesting target in cancer therapy. PMID- 11940466 TI - [Quality of life measurement in cancer survivors]. AB - The quality of life (QOL) of cancer patients can be measured by semi structured interview or by validated QOL instruments. Late effect toxicity scales can be used but do not account for the different domains of QOL measured by the patient. Health related quality of life can be measured by health general questionnaire or by specific cancer questionnaires. These instruments add to a core questionnaire, modules according for the organ or the toxicity of the treatments developed to evaluate cancer treatments. Some questions can be added to evaluate special domain as medical expenses, personal insurance. Some specific questionnaires for long-term survivors are available, which explore patient's satisfaction according to this status for different domains of QOL or quote different aspects in comparison with a normal population and are transformed into an utility index. There is no gold standard to measure long-term survivors' quality of life. The use of proxy is limited by discrepancies observed in the evaluation of pain, psychological symptoms and general quality of life. QOL can be evaluated in a cohort of patients in a longitudinally study or better in a case control study. Examples of QOL evaluation of different cancers sites are given. PMID- 11940467 TI - [Weekly administration of paclitaxel in the treatment of metastatic breast cancer: from rational to practice]. AB - Clinical studies showed that paclitaxel is active as single agent and in combination chemotherapy for the management of metastatic breast cancer. A variety of doses and administration schedules have been evaluated in phase II and III trials. Weekly administration of paclitaxel which is generally well tolerated increases the therapeutic index (efficacy/safety) resulting from anti-angiogenic and pro-apoptotic effects. PMID- 11940468 TI - [News in occupational cancers]. AB - Despite the knowledge of the link between many sites of cancer occurence and previous occupational exposure, occupational cancers are generally underestimated. These cancers can be prevented through specific plans. In France, the number of cases requesting and receiving compensation for occupational cancer is increasing. The frequency of occupational exposure to carcinogens was recently evaluated. Legislation was reinforced in order to obtain a better control of exposure to carcinogens in the workplace. Lung cancer is the most frequent of occupational cancers. Epidemiological studies contribute to a better knowledge of etiologies and occupations responsible for the outcome of these cancers and allow quantification of the risk of cancer linked to different situations of exposure. Beside classical epidemiological studies, molecular epidemiology aims at identifying molecular targets of occupational agents. This approach may allow a better knowledge of the part played by occupational agents in these multifactorial diseases. PMID- 11940469 TI - [Food and cancer: state of the art about the protective effect of fruits and vegetables]. AB - Epidemiological studies performed during the last 20 years support an inverse relationship between the individual intake of fruits and vegetables and the risk of cancer. In taking into account some recent conflicting data, a working group of the Nacre network, the French Network for Food and Cancer Research, has conduced a critical analysis of epidemiological and experimental studies, including the preliminary data from the Epic cohort, the European Prospective Investigation into Cancer and Nutrition, to clarify the role of fruits and vegetables to prevent cancer. To date, a high intake of fruits and vegetables (at least, 400 g per day) is appropriate to lower the risk of cancer. Fruits and vegetables provide numerous phytochemicals which, in part, may explain their beneficial effect. Thus, studies in animal models and in cell-culture systems have furnished a lot of information about the potential mechanism by which a diet high in fruits and vegetables may reduce the risk of cancer in humans. However, more investigation in the identification of the biologically active constituents, in the knowledge of their availability and the mechanism by which they contribute to lower the risk of cancer, will increase the scientific support of a public health policy. PMID- 11940470 TI - [Impact of [18F]-FDG-PET on medical decision making in oncology: evaluation by the referring physicians during the opening year]. AB - The first study evaluating directly by the referring physician the clinical impact of [18F]-FDG-PET on modification of patient's management was performed only recently in California by means of a questionnaire. We have used the same methodology to evaluate this clinical impact during the opening year of our PET centre in France. A questionnaire was sent to the referring physician of each of the 476 patients who had at least one routine FDG-PET examination during the year 2000. Of 348 responses (response rate = 73%), the disease was upstaged in 26% of the cases and down-staged in 9%. Intermodality management changes (change from a scheduled therapeutic modality for a different one) were reported in 37% of the cases and intramodality changes in 9%. Those modification rates were respectively 38% and 7% in recurrence of colorectal cancer (153 patients), 47% and 7% in lung cancer (118 patients), 16% and 23% in lymphoma (43 patients), 25% and 6% in the staging of head and neck cancers (32 patients). When comparing with the corresponding studies performed in California, there were no significant differences between the rates of intermodality management changes. In contrast, intramodality management changes were less frequent in our survey, except for lymphoma. Globally, the clinical impact of FDG PET was similar with a higher response rate in our study (73% versus 35%); it was above the mean rate derived from a recent meta-analysis in more than 5,000 patients. PMID- 11940471 TI - [Breast cancer: factors influencing the therapeutic itinerary of patients in a medical oncology unit in Bamako (Mali)]. AB - Early therapy is a determining factor to the recovery in patients with breast cancer. The situation in Mali is characterized by the delayed diagnosis of this cancer which raises the hypothesis that medical itinerary of patients received in specialized oncology unit is particular. In order to verify this hypothesis, 44 patients including 43 women and one man aged 25 to 80 years (mean age 46.0 19.6 years), seen in medical oncology unit in Point G, were subjects of an interview about the motivation of their therapeutic itinerary. 22.7% was initially seen by a traditional physician and 77.3% by a health care professional. The request of care was influenced by the patient's representation of the disease and by their neighboring. The therapeutic itinerary: "from traditional medicine to conventional medicine" was the more frequently observed in our patients with a long delay between the first consultation and the specialized one. Very few patients have received information about their illness before their specialized consultation. We conclude that the medical itinerary of our patients is particular, that this itinerary is influenced by the patient's representation of the cancer and by difficulty in the relationship between patients and health professional. This raise questions about the quality of both the communication and the provided health care. So, health care for patients with breast cancer in Mali might widely consider the anthropological dimension of the disease. PMID- 11940474 TI - The role of apoptosis in myelodysplastic syndrome. PMID- 11940475 TI - Human herpesvirus 8-associated primary effusion lymphoma in HIV--patients: a clinicopidemiologic variant resembling classic Kaposi's sarcoma. PMID- 11940476 TI - Is thalidomide a true anti-angiogenic molecule in multiple myeloma? PMID- 11940477 TI - Inside Haematologica: caution in the use of thalidomide for treatment of hematologic disorders. PMID- 11940478 TI - Characterization of CD34+, CD13+, CD33- cells, a rare subset of immature human hematopoietic cells. AB - BACKGROUND AND OBJECTIVES: Hematopoietic progenitor cells that express CD34 are heterogeneous in their lineage affiliation and degree of maturation. Expression of CD13 and CD33 antigens indicates myeloid lineage association, but the precise sequence of expression of these two markers during differentiation is unclear. We noted the presence of CD34+ cells expressing CD13 but lacking CD33, a subset of cells not yet well characterized. In this report we describe the prevalence and the immunophenotype of this cell subset. DESIGN AND METHODS: We studied the immunophenotype of immature myeloid cells in human bone marrow samples from 11 healthy transplantation donors and in 4 cord blood samples. We used four-color flow cytometry and a large panel of monoclonal antibodies directed against lineage and differentiation-associated antigens. Three additional bone marrow samples were analyzed after immunomagnetic sorting of CD34+ cells. We focused our analysis on the subset of cells defined by the expression of CD34 and CD13 and the lack of CD33. RESULTS: We found CD34+, CD13+, CD33- cells in all 11 bone marrow and 4 cord blood samples studied. These cells represented 0.5 0.5% (mean SD) and 0.8 1.2% of mononucleated cells, respectively. CD34+, CD13+, CD33- cells appeared to be more immature than those expressing CD33 because of their light scatter characteristics (smaller size and lower granularity), the expression of markers associated with early hematopoietic cells (CD90, CD133 and CD117), and the absence of lineage-associated markers. INTERPRETATION AND CONCLUSIONS: These findings suggest that the expression of CD13 precedes that of CD33 during myeloid differentiation, and that CD34+, CD13+, CD33- cells are at an early stage of human myeloid cell differentiation. PMID- 11940479 TI - Translocation t(2;7)(p12;q21-22) with dysregulation of the CDK6 gene mapping to 7q21-22 in a non-Hodgkin's lymphoma with leukemia. AB - BACKGROUND AND OBJECTIVES: A female patient presented with splenomegaly and lymphocytosis with atypical lymphoid cell morphology. We identified t(2;7)(p12;q21) prompting studies of the translocation breakpoint and its consequences on protein expression to confirm or otherwise the recently reported involvement of CDK6 and IG k genes in the t(2;7) leading to over-expression of CDK6 protein. DESIGN AND METHODS: A variety of clinical and laboratory techniques including cell marker, cytogenetic and histologic studies were applied in order to establish the diagnosis. Fluorescence in situ hybridization (FISH) and Southern blotting were used for mapping the translocation breakpoint and Western blotting for assessing protein expression. RESULTS: Immunophenotyping showed the presence of a B-cell population with strong expression of FMC7, CD22, CD79b, CD5 and k restricted surface immunoglobulins. Based on morphology and immunophenotypic markers the diagnosis of B-cell non-Hodgkin's lymphoma was made. Karyotyping revealed a clone with t(2;7)(p12;q21-22). Evidence for clonal evolution with additional abnormalities including a deletion of the TP53 was present. We established by FISH and Southern blotting that the breakpoint on 7q21 22 fell in a region 66kb telomeric to the previously reported breakpoint for the t(2;7) and was the same as that observed in a t(7;21). CDK6 protein was over expressed. The patient received alkylating agents and splenectomy and is alive but the lymphocytosis persists with evidence of disease progression. INTERPRETATIONS AND CONCLUSIONS: We have demonstrated that CDK6 expression is dysregulated even when the breakpoint on 7q21-22 is located 66kb upstream from the coding region. Interestingly, the precise assignment of the lymphoma type in our case was not possible even when the splenic histology was analyzed. PMID- 11940480 TI - Identification of chromosomal translocations in leukemias by hybridization with oligonucleotide microarrays. AB - BACKGROUND AND OBJECTIVES: Identification of chromosomal rearrangements is important for a precise risk-stratified diagnosis of hematologic malignancies. As the number of known translocations, specific for different types of leukemia increases, it takes ever more time and increasing amounts of patient's material to screen a single patient with individual polymerase chain reactions (PCR). The aim of this study was to develop a new approach combining specificity with high throughput sufficient for rapid screening of clinical samples for the presence of numerous translocations. DESIGN AND METHODS: We designed an oligonucleotide microarray and used hybridization with microarrays in combination with multiplex reverse transcription-polymerase chain reaction (RT-PCR) assay for accurate and rapid identification of some major leukemias. The following translocations were used as prototypic: t(9;22) p210 and p190, t(4;11), t(12;21), and t(15;17). This approach was tested on five different cell cultures carrying translocations and on 22 clinical samples from leukemic patients. RESULTS: Distinctive hybridization signals were obtained for all types of chimeric transcripts from cell lines with translocations. Both the type of translocation and the splice variant were determined. The data demonstrated high specificity and reproducibility of the method. Analysis of the 22 clinical samples using the microarray-based approach showed complete agreement with standard PCR analysis. INTERPRETATION AND CONCLUSIONS: Our data suggest that oligonucleotide microarrays can be used as an efficient, alternative approach to the traditional post-PCR Southern blot analysis. The oligonucleotide microarray approach appears suitable for clinical screening of major risk-stratifying translocations in patients with leukemia. PMID- 11940481 TI - Interleukin-11 induces proliferation of human T-cells and its activity is associated with downregulation of p27(kip1). AB - BACKGROUND AND OBJECTIVES: We have recently shown that interleukin (IL-)11 induces polarization of human T-cells by inhibiting macrophage production of IL 12 and by exerting a direct effect on CD4+ T-cells. In this study, we investigated the effects of IL-11 on the kinetic activation and apoptosis of T cell subsets stimulated with anti-CD3/CD28 antibodies, anti-CD3 and IL-2 or dendritic cells. DESIGN AND METHODS: Apoptosis and cell cycle analysis of T-cells were assessed by double staining with propidium iodide and intracellular Ki-67 and by acridine orange staining. The expression of the negative regulator of the cell cycle p27Kip1 (p27) was also determined by flow cytometry. RESULTS: Our results show that 18 hours of incubation with IL-11 resulted in a significantly higher number of cycling CD4+ cells, CD4+CD45RA+ naive T-cells and CD4+CD45RO+ memory T-cells, but not of CD8+ cells. The kinetic activity of IL-11 was observed up to 72 hours, when the peak value of S-phase cells occurred. IL-11 also significantly enhanced CD4+ and CD4+CD45RA+ cell proliferation when T-cells were co-incubated with allogeneic dendritic cells. Conversely, IL-11 did not protect any of the T-cell subsets from apoptosis. At the functional level, a type-2 cytokine pattern of cultured T-lymphocytes was observed after 5 days of incubation with IL-11. Proliferation and functional activation of T-cells were preceeded by downregulation of p27, which occurred as early as 12 hours after incubation with IL-11. INTERPRETATION AND CONCLUSIONS: IL-11 induces Th-2 polarization and cell-cycle entry of human CD4+, CD4+CD45RA+ and CD4+CD45RO+cells and their activation is associated with the downregulation of p27. PMID- 11940482 TI - What does apoptosis have to do with clinical features in myelodysplastic syndrome? AB - BACKGROUND AND OBJECTIVES: Apoptosis is thought to play a key pathogenic role in myelodysplastic syndrome. The aim of our study was to determine whether apoptotic index, p53 and bcl-2 levels correlate with the clinical consequences of ineffective hematopoiesis; namely, the development of cytopenia and the shortened survival of patients with myelodysplastic syndrome. DESIGN AND METHODS: The apoptotic index, determined by TdT-mediated dUTP nick-end labeling, as well as bcl-2 and p53 protein levels, evaluated by immunohistochemistry, were studied in bone marrow trephine biopsy archival samples from 25 healthy subjects aged over fifty (control group), 61 consecutive patients with myelodysplastic syndrome and 17 patients with de novo acute myeloid leukemia according to FAB criteria. The correlation between relevant clinical parameters was analyzed with Spearman's correlation test. Factors influencing survival were studied by Cox proportional hazards regression analysis. RESULTS: The apoptotic index did not correlate with blood counts at diagnosis and had no prognostic influence on the overall survival of patients with myelodysplastic syndrome. The p53 score correlated with both the leukocyte count (r=-0.274, p=0.034, Spearman's) and hemoglobin concentration (r= 0.316; p=0.014) in the myelodysplastic syndrome patients and showed an independent and significant prognostic influence on their overall survival (p=0.045, Cox's) while the bcl-2 score was not correlated with their blood counts or prognosis. INTERPRETATION AND CONCLUSIONS: Apoptotic index and bcl-2 do not correlate with key clinical data in patients with myelodysplastic syndrome, while p53 levels show a good correlation with these data and may be a useful parameter to add to current prognostic schemes in this entity. PMID- 11940483 TI - Essential thrombocythemia with ringed sideroblasts: a heterogeneous spectrum of diseases, but not a distinct entity. AB - BACKGROUND AND OBJECTIVES: According to the recently published WHO-classification essential thrombocythemia with ringed sideroblasts (ET/RS) remains an ambiguous category which may be considered as myelodysplastic/myeloproliferative disease, unclassifiable. Because until now only case reports or very small series of patients have been described, a more systematically performed study is warranted. DESIGN AND METHODS: A retrospective evaluation was carried out on 38 patients with the diagnosis of ET/RS and more than 15 % ringed sideroblasts on smears. Simultaneously performed bone marrow biopsies, follow-up examinations and survival data were also available. RESULTS: Based on cytological features and particular bone marrow findings including immunohistochemistry three patterns could be determined. These were associated with different clinical features and in particular prognosis. Group I included six patients whose diagnosis was consistent with ET, group II comprised 21 patients revealing prefibrotic and early fibrotic chronic idiopathic myelofibrosis (CIMF) and finally 11 patients (group III) displayed myelodysplastic syndromes (MDS). Follow-up studies revealed that no patient with ET showed a fiber increase but eight CIMF patients developed overt myelofibrosis and four patients of the MDS group developed secondary acute myeloid leukemia. In comparison with a control group of 39 patients with true ET, prognosis was significantly different because our cohort showed a median survival of 100 months that contrasted significantly with the 170 months in the patients with true ET. INTERPRETATION AND CONCLUSIONS: Ringed sideroblasts are not a pathognomonic feature of MDS, but may indicate a dysplasia probably associated with a primary or secondary disturbance of iron metabolism in a variety of disorders. For this reason, a more accurate classification of so-called ET/RS patients is warranted by evaluation of smears and in particular bone marrow biopsy specimens. According to our findings these patients should be classified as having either ET, CIMF or MDS and show a significantly different survival pattern. PMID- 11940484 TI - Anti-idiotypic vaccination in the treatment of low-grade B-cell lymphoma. AB - BACKGROUND AND OBJECTIVES: Patients with B-cell lymphoma can be induced to mount a specific immune response against the individual idiotypic determinants expressed in their tumor cells. This form of active immunotherapy is now under evaluation in the clinical setting. We evaluated the feasibility and effectiveness of this kind of immunotherapy in a group of patients with low-grade lymphoma, which included two cases of bi/triclonal lymphoma. DESIGN AND METHODS: Nine patients with a histopathologic diagnosis of follicular non-Hodgkin's (NHL) low-grade B-cell lymphoma were initially selected for this disease-free survival study. Idiotypic proteins were recovered by somatic fusion of the tumor cells and their identity with the tumor idiotype determined by molecular methods. The patients received the vaccine consisting of their tumor Ig protein coupled to keyhole limpet hemocyanine and were observed for toxicity, anti-idiotypic immune response, clinical outcome and circulating t(14;18)+ tumor cells. RESULTS: The median duration of follow-up was 40 (10-64) months from the initiation of immunotherapy. Tumor regression was detected in two patients. No tumor progression was observed in the other patients. Eight patients generated specific anti-idiotypic antibodies and 3 out of five were cleared of circulating t(14;18)+ cells. INTERPRETATION AND CONCLUSIONS: Induction of tumor-specific anti-idiotypic immune responses may be of benefit to patients affected by low-grade B-cell NHL. Our results are in line with those previously reported and call attention to the issue of tumor clonality in this kind of treatment. PMID- 11940486 TI - Thromboplastin-thrombomodulin-mediated time: a new global test sensitive to protein S deficiency and increased levels of factors II, V, VII and X. AB - BACKGROUND AND OBJECTIVES: A new test for screening the procoagulant capacity of plasma is described and evaluated. This test is based on the coagulation of plasma initiated by thromboplastin (Tp) in the presence of thrombomodulin (TM). In a previous paper we reported that this test had a significant phenotypic and genetic correlation with thrombosis susceptibility. The present report describes the characteristics of the test and its sensitivity to the concentration of some hemostasis factors. DESIGN AND METHODS: Plasma from normal subjects, from individuals with various disorders of hemostasis and plasma with different concentrations of factors II, V, VII, VIII, X, fibrinogen, protein C and protein S were studied. The thromboplastin-thrombomodulin-mediated time (Tp-TMT) is measured after mixing 100 mL of plasma diluted 1/10 at 37 C with 100 mL of a solution composed of 2 parts of thromboplastin, 1 part of thrombomodulin at 30 U/mL and 1 part of Owren's buffer. The results are expressed as the ratio of the patient's clotting time to that of the control. Values were compared with Student's t test and the Mann-Whitney test. Differences were considered statistically significant when p<0.05. RESULTS: In the control group women showed significantly lower values than men. Raised levels of factors II, V, VII and X reduced the coagulation time obtained with Tp-TM. Elevated concentrations of fibrinogen and factor VIII did not influence the test. The Tp-TMT was sensitive to protein S deficiencies, but not to protein C deficiencies. INTERPRETATION AND CONCLUSIONS: These results indicate that the effect of protein S on the test is through its anti-prothrombinase activity. IN CONCLUSION: Tp-TMT, which is correlated with thrombosis susceptibility, is sensitive to raised levels of factors II, V, VII and X, as well as to low levels of protein S, and may be an indicator of thrombosis risk. PMID- 11940485 TI - Salvage therapy with thalidomide in patients with advanced relapsed/refractory multiple myeloma. AB - BACKGROUND AND OBJECTIVES: Few therapeutic options are presently available for patients with multiple myeloma (MM) who relapse after autologous or allogeneic stem cell transplantation, or for patients who are refractory to conventional chemotherapy and not eligible for salvage high-dose therapy. Thalidomide, a glutamic acid derivative with anti-angiogenic properties, has been recently proposed as an effective therapy for patients with advanced refractory disease. The aim of this study was to evaluate the activity of thalidomide in a large series of MM patients. DESIGN AND METHODS: From October 1999 to January 2001, 65 patients (46 males/19 females) from 8 Italian institutions were treated with thalidomide. Twenty-six patients had relapsed after autologous stem cell transplantation, either single (n = 12) or double (n= 12); 38 patients had shown disease progression after >= 2 lines of conventional chemotherapy, 2 patients had relapsed after allotransplant, one single patient had not received previous treatment. Sixty-one (93.8%) patients were in stage III, median b2 microglobulin was 2.9 mg/L, and median bone marrow plasma cell infiltration was 50%. Thalidomide was initially administered at a dose of 100 mg/day; if well tolerated, the dose was to be increased serially by 200mg every other week to a maximum of 800 mg/day. RESULTS: The median administered dose of thalidomide was 400 mg/day. WHO grade > II toxic effects were constipation (52%), lethargy (34%), skin rash (11%), peripheral neuropathy (14%) and leukopenia (3%). Sixty patients are presently evaluable for response; of these, 17 (28.3%) showed > or = 50% reduction in serum or urinary M protein concentration and 11 (18.3%) showed > or = 25% tumor reduction, for a total response rate averaging 46.6%. After a median of 8 months' follow-up, 15/28 patients are alive and progression-free (at 2 to 16 months), 12 patients have relapsed, and 1 patient died of pulmonary edema while still in partial remission. Among pre-treatment variables that were analyzed for their potential relationship with tumor response, only the concentration of vascular endothelial growth factor (VEGF) in the conditioned media obtained upon culture of bone marrow plasma cells was statistically significant. Plasma cells from patients who responded favorably to thalidomide secreted a significantly lower amount of VEGF than plasma cells from resistant patients (126.45 165 pg/mL vs 227.11 70 pg/mL, p=0.04). INTERPRETATION AND CONCLUSIONS: These data confirm that thalidomide is active in patients with advanced relapsed/refractory MM and represent the basis for ongoing clinical trials aimed at testing the role of this drug as front line therapy for newly diagnosed disease. PMID- 11940487 TI - Vitamin E potentiates the antiplatelet activity of aspirin in collagen-stimulated platelets. AB - BACKGROUND AND OBJECTIVES: The combination of vitamin E with aspirin is becoming an attractive therapeutic approach to prevent thrombotic vascular accidents. In this study we investigated the capacity of vitamin E (50 and 100 M) to enhance the antiplatelet effect of aspirin. DESIGN AND METHODS: The dose-response curves of platelet aggregation, dense body secretion, phospholipase C activation and calcium mobilization were measured in aspirin-treated platelets with and without added vitamin E (50 and 100 M). The role of vitamin E in reducing platelet adhesion to collagen was also studied. RESULTS: We demonstrated that, in platelets incubated with 100 M vitamin E, collagen-concentration ( g/mL) able to induce 50% of the maximal platelet aggregation and of the calcium mobilization was higher than in controls (11.6 versus 3.8 and 21.3 versus 9.8, respectively). Furthermore, 50 M vitamin E reduced platelet adhesion to collagen by about 80%. INTERPRETATION AND CONCLUSIONS: These data demonstrate that vitamin E can potentiate the antiplatelet activity of aspirin by inhibiting the early events of platelet activation pathways induced by collagen. This finding provides a rationale for combining aspirin and vitamin E to prevent thrombotic complications in atherosclerotic patients. PMID- 11940488 TI - Do patient-related blood donors represent a threat to the safety of the blood supply? AB - BACKGROUND AND OBJECTIVES: Patient-related blood donors contribute to a significant proportion of the blood units collected in hospital-based blood banks. However, there is some concern on the safety of this kind of donation because of the possible existence of incentives for the donor to conceal deferrable risk factors, thus increasing the risk of donation within the window period of transfusion-transmitted infections. We tested the hypothesis that if patient-related blood donors are less safe than community ones, the former would display both a higher prevalence of viral markers and a predominance of undisclosed risk-factors with low social acceptability. DESIGN AND METHODS: Comparison of virus reactivity rates against hepatitis C virus (HCV), hepatitis B virus (HBV) and human immunodeficiency virus (HIV), and the associated risk factors, between patient-related and community donors who donated whole blood in our center during a five-year period. RESULTS: During the period under study 72,226 donors gave 149,944 whole blood units, of which 22,888 (15.3%) were provided by patient-related donors. There were 273 confirmed virus-reactive donations (15 anti-HIV, 148 anti-HCV and 110 HBsAg). The adjusted prevalence of virus reactivity was 19 (95% CI: 11-35) times higher in first-time donors than in repeat donors, 3.5 (95% CI: 2.3-4.1) times higher in donors > or = 30 years old than in younger ones, and 2.5 (95% CI: 1.9-3.2) times higher in patient-related donors than in community donors. With regard to deferrable risk-factors not disclosed at the time of donation, there was no significant difference between patient-related and community donors in the frequency of people who denied any risk-factor or who admitted intravenous drug use or high-risk sex. Past household contact with individuals having liver disease was significantly more frequent in patient-related donors than in community ones. INTERPRETATION AND CONCLUSIONS: Our results do not support the hypothesis that patient-related donors represent an increased risk of window-period donation because they conceal deferrable risk factors more frequently than community donors. PMID- 11940489 TI - The interleukin-12 and interleukin-12 receptor system in normal and transformed human B lymphocytes. AB - BACKGROUND AND OBJECTIVES: Interleukin-12 (IL-12) is a heterodimeric cytokine that induces interferon-g (IFN-g) production by natural killer and T-lymphocytes. IL-12 also activates human B-cells through the IL-12 receptor (IL-12R) complex. Here we review the expression and function of IL-12 and IL-12R in human B-cells and in their malignant counterparts. EVIDENCE AND INFORMATION SOURCES: The information provided derives from results both published and unpublished obtained in the laboratories of the Authors, and from a comprehensive review of all the pertinent articles published so far in Medline. STATE OF ART: The two components of the IL-12R, i.e. the b 1 and b 2 chains, were found to be constitutively expressed in human naive, germinal center and memory tonsil B-cells; however, only naive B-cells were activated following interaction with IL-2. Here we show that the IL-12Rb2 gene is not expressed in EBV-transformed normal B-lymphocytes and in Burkitt's lymphoma B-cell lines. IL-12 p35 and p40 transcripts were detected in all tonsil B-cell subsets, but only naive and memory B-cells produced IL-12. In this study, biosynthesis of IL-12 was investigated in tonsil B-cells, showing that the molecular weight of the mature heterodimeric IL-12 was similar to that of monocyte-derived IL-12, with minor differences possibly related to glycosylation. Finally, malignant B-cells from follicular and marginal zone lymphomas expressed IL-12 p35 and p40 transcripts, whereas only p35 mRNA was detected in mantle cell lymphoma. PERSPECTIVES: Taken together, the studies herein reviewed indicate that human B-cells, at variance with their murine counterparts, can produce IL-12 following CD40 ligation. IL-12 p35 and p40 transcripts are found in B-cells from different lymphoproliferative disorders, but the evidence that the cytokine is produced at the protein level is poor. IL 12R is expressed in the main human B-cell subsets, but it is functional only in naive B-cells. Finally, the failure of transformed B-cell lines to express IL 12Rb2 mRNA opens up new perspectives in the investigation of B-cell malignant transformation. PMID- 11940490 TI - The (--(SEA)) alpha-thalassemia (SEA) deletion ameliorates the clinical phenotype of beta(0)/beta(+) but not necessarily beta(0)/beta(0) thalassemia. PMID- 11940491 TI - Clinical characteristics of and risk factors for herpes zoster after hematopoietic stem cell transplantation. PMID- 11940492 TI - Polymerase chain reaction-based "pre-emptive" therapy with cidofovir for cytomegalovirus reactivation in allogeneic hematopoietic stem cells transplantation recipients: a prospective study. PMID- 11940493 TI - Acute myeloid leukemia in the elderly: evaluation of overall survival in 69 consecutive patients. PMID- 11940494 TI - Triosephosphate isomerase deficiency. genetic, enzymatic and metabolic characterization of a new case from Spain. PMID- 11940495 TI - Leukaemia cutis: clinical features and treatment strategies. PMID- 11940496 TI - Treatment of a secondary myelodysplastic syndrome after allogenic bone marrow transplantation using Mylotarg. PMID- 11940497 TI - Erythroblast island. PMID- 11940498 TI - Simultaneous occurrence of chronic lymphocytic leukaemia and acute lymphoblastic leukaemia. PMID- 11940499 TI - Subcapsular splenic rupture in a patient with chronic lymphocytic leukemia. PMID- 11940501 TI - Erythrophagocytosis by peripheral monocytes in Plasmodium falciparum malaria. PMID- 11940500 TI - Gigantic cutaneous lymphoma following Di Bella's therapy. PMID- 11940502 TI - Gut perforation in MALT lymphoma of colon. PMID- 11940503 TI - Isolated massive meningeal progression in aggressive non Hodgkin's lymphoma. PMID- 11940504 TI - Late relapse of childhood acute lymphoblastic leukemia and pcr-monitoring of minimal residual disease: how much time can elapse between "molecular" and clinical relapse? PMID- 11940505 TI - Autoimmune hypothyroidism and retinoic acid. PMID- 11940506 TI - TATA-Box polymorphism in the uridine diphosphate glucuronosyl transferase gene in Portuguese patients with clinical diagnosis of Gilbert Syndrome. PMID- 11940507 TI - Combined systemic and intrathecal chemotherapy plus radiotherapy in testicular lymphoma: a report of two cases. PMID- 11940508 TI - Mechanisms of normal and tumor-derived angiogenesis. AB - Often those diseases most evasive to therapeutic intervention usurp the human body's own cellular machinery or deregulate normal physiological processes for propagation. Tumor-induced angiogenesis is a pathological condition that results from aberrant deployment of normal angiogenesis, an essential process in which the vascular tree is remodeled by the growth of new capillaries from preexisting vessels. Normal angiogenesis ensures that developing or healing tissues receive an adequate supply of nutrients. Within the confines of a tumor, the availability of nutrients is limited by competition among actively proliferating cells, and diffusion of metabolites is impeded by high interstitial pressure (Jain RK. Cancer Res 47: 3039-3051, 1987). As a result, tumor cells induce the formation of a new blood supply from the preexisting vasculature, and this affords tumor cells the ability to survive and propagate in a hostile environment. Because both normal and tumor-induced neovascularization fulfill the essential role of satisfying the metabolic demands of a tissue, the mechanisms by which cancer cells stimulate pathological neovascularization mimic those utilized by normal cells to foster physiological angiogenesis. This review investigates mechanisms of tumor-induced angiogenesis. The strategies used by cancer cells to develop their own blood supply are discussed in relation to those employed by normal cells during physiological angiogenesis. With an understanding of blood vessel growth in both normal and abnormal settings, we are better suited to design effective therapeutics for cancer. PMID- 11940509 TI - From genetics to cellular physiology. Focus on "Regulation of transferrin-induced endocytosis by wild-type and C282Y-mutant HFE in transfected HeLa cells". PMID- 11940510 TI - Regulation of transferrin-induced endocytosis by wild-type and C282Y-mutant HFE in transfected HeLa cells. AB - The hereditary hemochromatosis protein HFE is known to complex with the transferrin receptor; however, its function regarding endocytosis of transferrin is unclear. We performed patch-clamp capacitance measurements in transfected HeLa cells carrying wild-type or C282Y-mutant HFE cDNA under the control of a tetracycline-sensitive promoter. Whole cell experiments in cells with suppressed expression of wild-type HFE revealed a decrease in membrane capacitance, reflecting predominance of endocytosis in the presence of transferrin. Cells overexpressing C282Y-mutant HFE displayed less intense capacitance decreases, whereas no significant decrease was observed in cells overexpressing wild-type HFE. The formation of single endocytic vesicles in cells with suppressed expression of wild-type HFE was greatly increased in the presence of transferrin as revealed by cell-attached recordings. According to their calculated diameters, many of these vesicles corresponded to clathrin-coated vesicles. These results suggest that wild-type HFE negatively modulates the endocytic uptake of transferrin. This inhibitory effect is attenuated in cells expressing C282Y mutant HFE. Time-resolved measurements of cell membrane capacitance provide a powerful tool to study transferrin-induced endocytosis in single cells. PMID- 11940512 TI - Focus on "Cholangiocytes exhibit dynamic, actin-dependent apical membrane turnover". PMID- 11940511 TI - Beta-adrenergic receptors and Ca(2+). Focus on "Beta-adrenergic potentiation of endoplasmic reticulum Ca(2+) release in brown fat cells". PMID- 11940513 TI - Calcineurin differentially regulates maintenance and growth of phenotypically distinct muscles. AB - Adequate muscle mass is critical for human health. The molecular pathways regulating maintenance and growth of adult skeletal muscle are little understood. Calcineurin (CN) is implicated as a key signaling molecule in hypertrophy. Whether CN is involved in all forms of muscle growth or in different muscles is unknown. Here, we examine the role of CN in regulating maintenance of muscle size and growth of atrophied muscle in the soleus (slow) and plantaris (fast). The CN inhibitor cyclosporin A (CsA) differentially affects muscle growth and maintenance depending on muscle phenotype. The plantaris is more severely affected by CsA than the soleus in both growth conditions. One-week vs. 2-wk CsA treatment suggests that both CN-dependent and CN-independent growth occur in the atrophied soleus, whereas plantaris growth appears to be totally CN dependent. Our results suggest that CN regulates multiple types of muscle growth, depending both on muscle phenotype and stage of myofiber growth. Differential expression of components of the CN pathway occurs and may contribute to the differences between muscles. PMID- 11940514 TI - Myostatin is an inhibitor of myogenic differentiation. AB - Myostatin (MSTN), a transforming growth factor (TGF)-beta superfamily member, has been shown to negatively regulate muscle growth by inhibiting muscle precursor cell proliferation. Here, we stably transfected C(2)C(12) cells with mouse MSTN cDNA to investigate its possible role in myoblast differentiation. We found that MSTN cDNA overexpression reversibly inhibits the myogenic process by downregulating mRNA levels of the muscle regulatory factors myoD and myogenin, as well as the activity of their downstream target creatine kinase. Taking into consideration that MSTN expression during development is restricted to muscle, our results suggest that MSTN probably regulates myogenic differentiation by an autocrine mechanism. PMID- 11940515 TI - Regulation of intracellular calcium in N1E-115 neuroblastoma cells: the role of Na(+)/Ca(2+) exchange. AB - In fura 2-loaded N1E-115 cells, regulation of intracellular Ca(2+) concentration ([Ca(2+)](i)) following a Ca(2+) load induced by 1 microM thapsigargin and 10 microM carbonylcyanide p-trifluoromethyoxyphenylhydrazone (FCCP) was Na(+) dependent and inhibited by 5 mM Ni(2+). In cells with normal intracellular Na(+) concentration ([Na(+)](i)), removal of bath Na(+), which should result in reversal of Na(+)/Ca(2+) exchange, did not increase [Ca(2+)](i) unless cell Ca(2+) buffer capacity was reduced. When N1E-115 cells were Na(+) loaded using 100 microM veratridine and 4 microg/ml scorpion venom, the rate of the reverse mode of the Na(+)/Ca(2+) exchanger was apparently enhanced, since an approximately 4- to 6-fold increase in [Ca(2+)](i) occurred despite normal cell Ca(2+) buffering. In SBFI-loaded cells, we were able to demonstrate forward operation of the Na(+)/Ca(2+) exchanger (net efflux of Ca(2+)) by observing increases (approximately 6 mM) in [Na(+)](i). These Ni(2+) (5 mM)-inhibited increases in [Na(+)](i) could only be observed when a continuous ionomycin induced influx of Ca(2+) occurred. The voltage-sensitive dye bis-(1,3 diethylthiobarbituric acid) trimethine oxonol was used to measure changes in membrane potential. Ionomycin (1 microM) depolarized N1E-115 cells (approximately 25 mV). This depolarization was Na(+) dependent and blocked by 5 mM Ni(2+) and 250-500 microM benzamil. These data provide evidence for the presence of an electrogenic Na(+)/Ca(2+) exchanger that is capable of regulating [Ca(2+)](i) after release of Ca(2+) from cell stores. PMID- 11940517 TI - Beta-adrenergic potentiation of endoplasmic reticulum Ca(2+) release in brown fat cells. AB - We find that the adrenergic agonist isoproterenol increases intracellular Ca(2+) concentration ([Ca(2+)](i)) in cultured rat brown adipocytes. At the concentration used (10 microM), isoproterenol-induced Ca(2+) responses were sensitive to block by either alpha(1)- or beta-adrenergic antagonists, suggesting an interaction between these receptor subtypes. Despite reliance on beta adrenoceptor activation, the Ca(2+) response was not due solely to increases in cAMP because, administered alone, the selective beta(3)-adrenergic agonist BRL 37344 or forskolin did not increase [Ca(2+)](i). However, increased cAMP elicited vigorous [Ca(2+)](i) increases in the presence of barely active concentrations of the alpha-adrenergic agonist phenylephrine or the P2Y receptor agonist UTP. Consistent with isoproterenol recruiting only inositol 1,4,5-trisphosphate (IP(3))-sensitive Ca(2+) stores, endoplasmic reticulum store depletion by thapsigargin blocked isoproterenol-induced Ca(2+) increases, but removal of external Ca(2+) did not. These results argue that increases in cAMP sensitize the IP(3)-mediated Ca(2+) release system in brown adipocytes. PMID- 11940516 TI - Mechanism of constrictive vascular remodeling by homocysteine: role of PPAR. AB - To test the hypothesis that homocysteine induces constrictive vascular remodeling by inactivating peroxisome proliferator-activated receptor (PPAR), aortic endothelial cells (ECs) and smooth muscle cells (SMCs) were isolated. Collagen gels were prepared, and ECs or SMCs (10(5)) or SMCs + ECs (10(4)) were incorporated into the gels. To characterize PPAR, agonists of PPAR-alpha [ciprofibrate (CF)] and PPAR-gamma [15-deoxy-12,14-prostaglandin J(2) (PGJ(2))] were used. To determine the role of disintegrin metalloproteinase (DMP), cardiac inhibitor of metalloproteinase (CIMP) was used in collagen gels. Gel diameter at 0 h was 14.1 +/- 0.2 mm and was unchanged up to 24 h as measured by a digital micrometer. SMCs reduce gel diameter to 10.5 +/- 0.4 mm at 24 h. Addition of homocysteine to SMCs reduces further the gel diameter to 8.0 +/- 0.2 mm, suggesting that SMCs induce contraction and that the contraction is further enhanced by homocysteine. Addition of ECs and SMCs reduces gel diameter to 12.0 +/- 0.3 mm, suggesting that ECs play a role in collagen contraction. Only PGJ(2), not CF, inhibits SMC contraction. However, both PGJ(2) and CF inhibit contraction of ECs and SMCs + ECs. Addition of anti-DMP blocks SMC- as well as homocysteine mediated contraction. However, CIMP inhibits only homocysteine-mediated contraction. The results suggest that homocysteine may enhance vascular constrictive remodeling by inactivating PPAR-alpha and -gamma in ECs and PPAR gamma in SMCs. PMID- 11940518 TI - Time-dependent changes in expression of troponin subunit isoforms in unloaded rat soleus muscle. AB - This study focuses on the effects of mechanical unloading of rat soleus muscle on the isoform patterns of the three troponin (Tn) subunits: troponin T (TnT), troponin I (TnI), and troponin C (TnC). Mechanical unloading was achieved by hindlimb unloading (HU) for time periods of 7, 15, and 28 days. Relative concentrations of slow and fast TnT, TnI, and TnC isoforms were assessed by electrophoretic and immunoblot analyses. HU induced profound slow-to-fast isoform transitions of all Tn subunits, although to different extents and with different time courses. The effectiveness of the isoform transitions was higher for TnT than for TnI and TnC. Indeed, TnI and TnC encompassed minor partial exchanges of slow isoforms with their fast counterparts, whereas the expression pattern of fast TnT isoforms (TnTf) was largely increased after HU. Moreover, slow and fast isoforms of the different Tn were not affected in the same manner by HU. This suggests that the slow and fast counterparts of the Tn subunit isoforms are regulated independently in response to HU. The changes in TnTf composition occurred in parallel with previously demonstrated transitions within the pattern of the fast myosin heavy chains in the same muscles. PMID- 11940519 TI - Human Na(+)/H(+) exchanger isoform 6 is found in recycling endosomes of cells, not in mitochondria. AB - Since the discovery of the first intracellular Na(+)/H(+) exchanger in yeast, Nhx1, multiple homologs have been cloned and characterized in plants. Together, studies in these organisms demonstrate that Nhx1 is located in the prevacuolar/vacuolar compartment of cells where it sequesters Na(+) into the vacuole, regulates intravesicular pH, and contributes to vacuolar biogenesis. In contrast, the human homolog of Nhx1, Na(+)/H(+) exchanger isoform 6 (NHE6), has been reported to localize to mitochondria when transiently expressed as a fusion with green fluorescent protein. This result warrants reevaluation because it conflicts with predictions from phylogenetic analyses. Here we demonstrate that when epitope-tagged NHE6 is transiently expressed in cultured mammalian cells, it does not colocalize with mitochondrial markers. It also does not colocalize with markers of the lysosome, late endosome, trans-Golgi network, or Golgi cisternae. Rather, NHE6 is distributed in recycling compartments and transiently appears on the plasma membrane. These results suggest that, like its homologs in yeast and plants, NHE6 is an endosomal Na(+)/H(+) exchanger that may regulate intravesicular pH and volume and contribute to lysosomal biogenesis. PMID- 11940520 TI - Cholangiocytes exhibit dynamic, actin-dependent apical membrane turnover. AB - The present studies of cholangiocytes used complementary histological, biochemical, and electrophysiological methods to identify a dense population of subapical vesicles, quantify the rates of vesicular trafficking, and assess the contribution of the actin cytoskeleton to membrane trafficking. FM 1-43 fluorescence measured significant basal rates of total exocytosis (1.33 +/- 0.16% plasma membrane/min) in isolated cholangiocytes and apical exocytosis in cholangiocyte monolayers. Cell surface area remained unchanged, indicating that there was a concurrent, equivalent rate of endocytosis. FM 1-43 washout studies showed that 36% of the endocytosed membrane was recycled to the plasma membrane. 8-(4-Chlorophenylthio)adenosine 3',5'-cyclic monophosphate (CPT-cAMP; cAMP analog) increased exocytosis by 71 +/- 31%, whereas the Rp diastereomer of adenosine 3',5'-cyclic monophosphothioate (Rp-cAMPS; protein kinase A inhibitor) diminished basal exocytosis by 53 +/- 11%. A dense population of 140-nm subapical vesicles arose, in part, from apical membrane endocytosis. Phalloidin staining showed that a subpopulation of the endocytosed vesicles was encapsulated by F actin. Furthermore, membrane trafficking was inhibited by disrupting the actin cytoskeleton with cytochalasin D (51 +/- 13% of control) or jasplakinolide (58 +/ 9% of control). These studies indicate that there is a high rate of vesicular trafficking at the apical membrane of cholangiocytes and suggest that both cAMP and the actin cytoskeleton contribute importantly to these events. PMID- 11940522 TI - H(+)-linked transport of salicylic acid, an NSAID, in the human trophoblast cell line BeWo. AB - We investigated the transport of salicylic acid and L-lactic acid across the placenta using the human trophoblast cell line BeWo. We performed uptake experiments and measured the change in intracellular pH (pH(i)). The uptakes of [(14)C]salicylic acid and L-[(14)C]lactic acid were temperature- and extracellular pH-dependent and saturable at higher concentrations. Both uptakes were also reduced by FCCP, nigericin, and NaN(3). Various nonsteroidal anti inflammatory drugs (NSAIDs) strongly inhibited the uptake of L-[(14)C]lactic acid. Salicylic acid and ibuprofen noncompetitively inhibited the uptake of L [(14)C]lactic acid. alpha-Cyano-4-hydroxycinnamate (CHC), a monocarboxylate transporter inhibitor, suppressed the uptake of L-[(14)C]lactic acid but not that of [(14)C]salicylic acid. CHC also suppressed the decrease of pH(i) induced by L lactic acid but had little effect on that induced by salicylic acid or diclofenac. These results suggest that NSAIDs are potent inhibitors of lactate transporters, although they are transported mainly by a transport system distinct from that for L-lactic acid. PMID- 11940521 TI - VEGF-induced mobilization of caveolae and increase in permeability of endothelial cells. AB - Glomerular epithelial cells (GEC) are a known site of vascular endothelial growth factor (VEGF) production. We established immortalized rat GEC, which retained the ability to produce VEGF. The isoforms expressed by GEC were defined as VEGF-205, 188, -120, and -164. The electrical resistance of endothelial cells cultured on GEC-conditioned matrix, an indicator of the permeability of monolayers to solutes, was significantly increased by the treatment with the neutralizing polyclonal antibodies to VEGF and decreased by VEGF-165. Transfection of endothelial cells with green fluorescence protein-caveolin construct and intravital confocal microscopy showed that VEGF results in a rapid appearance of transcellular elongated structures decorated with caveolin. Transmission electron microscopy of endothelial cells showed that caveolae undergo rapid internalization and fusion 30 min after application of VEGF-165. Later (36 h), endothelial cells pretreated with VEGF developed fenestrae and showed a decrease in electrical resistance. Immunoelectron microscopy of glomeruli confirmed VEGF localization to podocytes and in the basement membrane. In summary, immortalized GEC retain the ability to synthesize VEGF. Matrix-deposited and soluble VEGF leads to the enhancement of caveolae expression, their fission and fusion, formation of elongated caveolin-decorated structures, and eventual formation of fenestrae, both responsible for the increase in endothelial permeability. PMID- 11940523 TI - Excess plasma membrane and effects of ionic amphipaths on mechanics of outer hair cell lateral wall. AB - The interaction between the outer hair cell (OHC) lateral wall plasma membrane and the underlying cortical lattice was examined by a morphometric analysis of cell images during cell deformation. Vesiculation of the plasma membrane was produced by micropipette aspiration in control cells and cells exposed to ionic amphipaths that alter membrane mechanics. An increase of total cell and vesicle surface area suggests that the plasma membrane possesses a membrane reservoir. Chlorpromazine (CPZ) decreased the pressure required for vesiculation, whereas salicylate (Sal) had no effect. The time required for vesiculation was decreased by CPZ, indicating that CPZ decreases the energy barrier required for vesiculation. An increase in total volume is observed during micropipette aspiration. A deformation-induced increase in hydraulic conductivity is also seen in response to micropipette-applied fluid jet deformation of the lateral wall. Application of CPZ and/or Sal decreased this strain-induced hydraulic conductivity. The impact of ionic amphipaths on OHC plasma membrane and lateral wall mechanics may contribute to their effects on OHC electromotility and hearing. PMID- 11940524 TI - Estrogen pretreatment protects males against hypoxia-induced immune depression. AB - Hypoxemia depresses cell-mediated immune functions in males, whereas proestrous females do not show such a depression. We hypothesized that elevated systemic estradiol levels in proestrous females prevent hypoxemia-induced immune depression. To study this hypothesis, male C3H/HeN mice were pretreated with 17 beta-estradiol (E(2), 40 microg/kg body wt sc) or vehicle for 3 days before induction of hypoxemia and again immediately before induction of hypoxia. The mice were subjected to hypoxemia (95% N(2)-5% O(2)) or sham hypoxemia (room air) for 60 min, and plasma and spleen cells were collected 2 h later. In vehicle treated mice, splenocyte proliferation and interleukin-2 and interleukin-3 production were depressed after hypoxemia. E(2)-pretreated animals, however, displayed no such depression in splenic T cell parameters after hypoxemia. Splenic macrophage cytokine production was also depressed in vehicle-treated mice subjected to hypoxia, whereas it was normal in E(2)-pretreated mice. In summary, these findings indicate that administration of E(2) before hypoxemia prevented the depression of cell-mediated immune functions. Thus administration of 17 beta estradiol in high-risk patients before major surgery might decrease hypoxemia induced immune depression under those conditions. PMID- 11940525 TI - Shortening velocity and myosin heavy- and light-chain isoform mRNA in rabbit arterial smooth muscle cells. AB - In smooth muscle cells (SMCs) isolated from rabbit carotid, femoral, and saphenous arteries, relative myosin isoform mRNA levels were measured in RT-PCR to test for correlations between myosin isoform expression and unloaded shortening velocity. Unloaded shortening velocity and percent smooth muscle myosin heavy chain 2 (SM2) and myosin light chain 17b (MLC(17b)) mRNA levels were not significantly different in single SMCs isolated from the luminal and adluminal regions of the carotid media. Saphenous artery SMCs shortened significantly faster (P < 0.05) than femoral SMCs and had more SM2 mRNA (P < 0.05) than carotid SMCs and less MLC(17b) mRNA (P < 0.001) and higher tissue levels of SMB mRNA (P < 0.05) than carotid and femoral SMCs. No correlations were found between percent SM2 and percent MLC(17b) mRNA levels and unloaded shortening velocity in SMCs from these arteries. We have previously shown that myosin heavy chain (MHC) SM1/SM2 and SMA/SMB and MLC(17a)/MLC(17b) isoform mRNA levels correlate with protein expression for these isoforms in rabbit smooth muscle tissues. Thus we interpret these results to suggest that 1) SMC myosin isoform expression and unloaded shortening velocity do not vary with distance from the lumen of the carotid artery but do vary in arteries located longitudinally within the arterial tree, 2) MHC SM1/SM2 and/or MLC(17a)/MLC(17b) isoform expression does not correlate with unloaded shortening velocity, and 3) intracellular expression of the MHC SM1/SM2 and MLC(17a)/MLC(17b) isoforms is not coregulated. PMID- 11940526 TI - CLIC1 inserts from the aqueous phase into phospholipid membranes, where it functions as an anion channel. AB - CLIC1 is a member of the CLIC family of proteins, which has been shown to demonstrate chloride channel activity when reconstituted in phospholipid vesicles. CLIC1 exists in cells as an integral membrane protein and as a soluble cytoplasmic protein, implying that CLIC1 might cycle between membrane-inserted and soluble forms. CLIC1 was purified and detergent was removed, yielding an aqueous solution of essentially pure protein. Pure CLIC1 was mixed with vesicles, and chloride permeability was assessed with a chloride efflux assay and with planar lipid bilayer techniques. Soluble CLIC1 confers anion channel activity to preformed membranes that is indistinguishable from the previously reported activity resulting from reconstitution of CLIC1 into membranes by detergent dialysis. The activity is dependent on the amount of CLIC1 added, appears rapidly on mixing of protein and lipid, is inhibited by indanyloxyacetic acid-94, N ethylmaleimide, and glutathione, is inactivated by heat, and shows sensitivity to pH and to membrane lipid composition. We conclude that CLIC1 in the absence of detergent spontaneously inserts into preformed membranes, where it can function as an anion-selective channel. PMID- 11940527 TI - Beneficial effect of taurine depletion on osmotic sodium and calcium loading during chemical hypoxia. AB - Cellular sodium excess is cytotoxic because it increases both the intracellular osmotic load and intracellular calcium concentration ([Ca(2+)](i)). Because sodium levels rise during hypoxia, it is thought to contribute to hypoxic injury. Thus the present study tested the hypothesis that taurine-linked reductions in [Na(+)](i) reduce hypoxia-induced cell injury. Taurine depletion was achieved by exposing isolated neonatal cardiomyocytes to medium containing the taurine analog beta-Alanine. As predicted, the beta-Alanine-treated cell exhibited less hypoxia induced necrosis and apoptosis than the control, as evidenced by less swelling, shrinkage, TdT-mediated dUTP nick end labeling staining, and accumulation of trypan blue. After 1 h of chemical hypoxia, [Na(+)](i) was 3.5-fold greater in the control than the taurine-deficient cell. Although more taurine was lost from the control cell than from the beta-Alanine-treated cell during hypoxia, the combined taurine and sodium osmotic load was lower in the beta-Alanine-treated cell. Taurine deficiency also reduced the degree of hypoxia-induced calcium overload. Thus the observed resistance against hypoxia-induced necrosis and apoptosis is probably related to an improvement in sodium and calcium handling. PMID- 11940528 TI - Nonselective cation conductance activated by muscarinic and purinergic receptors in rat spiral ganglion neurons. AB - The present study characterizes the ionic conductances activated by acetylcholine (ACh) and ATP, two candidate neuromodulators, in isolated spiral ganglion neurons (SGNs). Brief application (1 s) of ACh evoked in a dose-dependent manner (EC(50) = 4.1 microM) a reversible inward current with a long latency (average 1.3 s), at holding potential (V(h)) = -50 mV. This current was reversibly blocked by atropine and mimicked by muscarine. Application of ATP also evoked a reversible inward current at V(h) = -50 mV, but the current showed two components. A fast component with a short latency was largely reduced when N-methyl-D-glucamine (NMDG) replaced extracellular sodium, implying a P2X-like ionotropic conductance. The second component had a longer latency (average 1.1 s) and was presumably activated by metabotropic P2Y-like receptors. The second component of ATP-evoked current shared similar characteristics with the responses evoked by ACh: the current reversed near 0 mV, displayed inward rectification, could be carried by NMDG, and was insensitive to extracellular and intracellular calcium. This ACh /ATP-evoked conductance was reversibly inhibited by preapplication of ionomycin. These results suggest that muscarinic receptors and purinergic metabotropic receptors activate a similar large nonselective cation conductance via a common intracellular pathway in SGNs, a candidate mechanism to regulate neuronal excitability of SGNs. PMID- 11940529 TI - Contribution of Na(+)-K(+)-Cl(-) cotransporter to high-[K(+)](o)- induced swelling and EAA release in astrocytes. AB - We hypothesized that high extracellular K(+) concentration ([K(+)](o))-mediated stimulation of Na(+)-K(+)-Cl(-) cotransporter isoform 1 (NKCC1) may result in a net gain of K(+) and Cl(-) and thus lead to high-[K(+)](o)-induced swelling and glutamate release. In the current study, relative cell volume changes were determined in astrocytes. Under 75 mM [K(+)](o,) astrocytes swelled by 20.2 +/- 4.9%. This high-[K(+)](o)-mediated swelling was abolished by the NKCC1 inhibitor bumetanide (10 microM, 1.0 +/- 3.1%; P < 0.05). Intracellular (36)Cl(-) accumulation was increased from a control value of 0.39 +/- 0.06 to 0.68 +/- 0.05 micromol/mg protein in response to 75 mM [K(+)](o). This increase was significantly reduced by bumetanide (P < 0.05). Basal intracellular Na(+) concentration ([Na(+)](i)) was reduced from 19.1 +/- 0.8 to 16.8 +/- 1.9 mM by bumetanide (P < 0.05). [Na(+)](i) decreased to 8.4 +/- 1.0 mM under 75 mM [K(+)](o) and was further reduced to 5.2 +/- 1.7 mM by bumetanide. In addition, the recovery rate of [Na(+)](i) on return to 5.8 mM [K(+)](o) was decreased by 40% in the presence of bumetanide (P < 0.05). Bumetanide inhibited high-[K(+)](o) induced (14)C-labeled D-aspartate release by ~50% (P < 0.05). These results suggest that NKCC1 contributes to high-[K(+)](o)-induced astrocyte swelling and glutamate release. PMID- 11940531 TI - Rapid and dynamic regulation of TGF-beta receptors on blood vessels and fibroblasts during ischemia-reperfusion injury. AB - The pathophysiological mechanisms involved in ischemia-reperfusion injury are poorly understood. Although transforming growth factor (TGF)-beta has been shown to provide protection against ischemia-reperfusion injury in different organ systems, little is known about the regulation of TGF-beta action during this process. Here we analyzed the effect of ischemia and reperfusion on the expression of TGF-beta and its receptors in vivo with a pig skin flap model. Analysis of unoperated skin, nonischemic control flap, ischemic flap, and reperfused flap by immunohistochemistry indicates that ischemia and reperfusion result in rapid and dynamic regulation of type I, II, and III TGF-beta receptors and TGF-beta1 in a cell type-specific manner. Furthermore, hypoxia upregulates type II TGF-beta receptor mRNA in skin fibroblasts in culture. Together, our results reveal that TGF-beta receptors and TGF-beta1 are markedly increased under acute ischemic conditions in the blood vessels and fibroblasts of the skin. We conclude that TGF-beta action is enhanced under ischemic conditions and that it may represent an adaptive response to ischemic injury. The augmented TGF-beta responsiveness may be a critical determinant of the protective effect of TGF-beta during ischemia-reperfusion injury. PMID- 11940530 TI - Astrocytes from Na(+)-K(+)-Cl(-) cotransporter-null mice exhibit absence of swelling and decrease in EAA release. AB - We reported previously that inhibition of Na(+)-K(+)-Cl(-) cotransporter isoform 1 (NKCC1) by bumetanide abolishes high extracellular K(+) concentration ([K(+)](o))-induced swelling and intracellular Cl(-) accumulation in rat cortical astrocytes. In this report, we extended our study by using cortical astrocytes from NKCC1-deficient (NKCC1(-/-)) mice. NKCC1 protein and activity were absent in NKCC1(-/-) astrocytes. [K(+)](o) of 75 mM increased NKCC1 activity approximately fourfold in NKCC1(+/+) cells (P < 0.05) but had no effect in NKCC1(-/-) astrocytes. Intracellular Cl(-) was increased by 70% in NKCC1(+/+) astrocytes under 75 mM [K(+)](o) (P < 0.05) but remained unchanged in NKCC1(-/-) astrocytes. Baseline intracellular Na(+) concentration ([Na(+)](i)) in NKCC1(+/+) astrocytes was 19.0 +/- 0.5 mM, compared with 16.9 +/- 0.3 mM [Na(+)](i) in NKCC1(-/-) astrocytes (P < 0.05). Relative cell volume of NKCC1(+/+) astrocytes increased by 13 +/- 2% in 75 mM [K(+)](o), compared with a value of 1.0 +/- 0.5% in NKCC1(-/-) astrocytes (P < 0.05). Regulatory volume increase after hypertonic shrinkage was completely impaired in NKCC1(-/-) astrocytes. High-[K(+)](o)-induced (14)C labeled D-aspartate release was reduced by approximately 30% in NKCC1(-/-) astrocytes. Our study suggests that stimulation of NKCC1 is required for high [K(+)](o)-induced swelling, which contributes to glutamate release from astrocytes under high [K(+)](o). PMID- 11940532 TI - Domain-domain associations in cystic fibrosis transmembrane conductance regulator. AB - Cystic fibrosis is caused by mutations in the cystic fibrosis transmembrane conductance regulator (CFTR) gene. CFTR is a chloride channel whose activity requires protein kinase A-dependent phosphorylation of an intracellular regulatory domain (R-domain) and ATP hydrolysis at the nucleotide-binding domains (NBDs). To identify potential sites of domain-domain interaction within CFTR, we expressed, purified, and refolded histidine (His)- and glutathione-S-transferase (GST)-tagged cytoplasmic domains of CFTR. ATP-binding to his-NBD1 and his-NBD2 was demonstrated by measuring tryptophan fluorescence quenching. Tryptic digestion of in vitro phosphorylated his-NBD1-R and in situ phosphorylated CFTR generated the same phosphopeptides. An interaction between NBD1-R and NBD2 was assayed by tryptophan fluorescence quenching. Binding among all pairwise combinations of R-domain, NBD1, and NBD2 was demonstrated with an overlay assay. To identify specific sites of interaction between domains of CFTR, an overlay assay was used to probe an overlapping peptide library spanning all intracellular regions of CFTR with his-NBD1, his-NBD2, and GST-R-domain. By mapping peptides from NBD1 and NBD2 that bound to other intracellular domains onto crystal structures for HisP, MalK, and Rad50, probable sites of interaction between NBD1 and NBD2 were identified. Our data support a model where NBDs form dimers with the ATP-binding sites at the domain-domain interface. PMID- 11940533 TI - Involvement of human PECAM-1 in angiogenesis and in vitro endothelial cell migration. AB - Platelet endothelial cell adhesion molecule (PECAM)-1 has been implicated in angiogenesis, but a number of issues remain unsettled, including the independent involvement of human PECAM-1 (huPECAM-1) in tumor angiogenesis and the mechanisms of its participation in vessel formation. We report for tumors grown in human skin transplanted on severe combined immunodeficiency mice that antibodies against huPECAM-1 (without simultaneous treatment with anti-VE-cadherin antibody) decreased the density of human, but not murine, vessels associated with the tumors. Anti-huPECAM-1 antibody also inhibited tube formation by human umbilical vein endothelial cells (HUVEC) and the migration of HUVEC through Matrigel-coated filters or during the repair of wounded cell monolayers. The involvement of huPECAM-1 in these processes was confirmed by the finding that expression of huPECAM-1 in cellular transfectants induced tube formation and enhanced cell motility. These data provide evidence of a role for PECAM-1 in human tumor angiogenesis (independent of VE-cadherin) and suggest that during angiogenesis PECAM-1 participates in adhesive and/or signaling phenomena required for the motility of endothelial cells and/or their subsequent organization into vascular tubes. PMID- 11940535 TI - Why are some individuals resistant to the cardioprotective effects of aspirin? Could it be thromboxane A2? PMID- 11940536 TI - Accelerated reendothelialization with suppressed thrombogenic property and neointimal hyperplasia of rabbit jugular vein grafts by adenovirus-mediated gene transfer of C-type natriuretic peptide. AB - BACKGROUND: Vein graft disease limits the late results of coronary revascularization. C-type natriuretic peptide (CNP) inhibits the growth of vascular smooth muscle cells. Given the effects of CNP on cGMP cascade, we hypothesized that transfected CNP genes modulate endothelial repair and thrombogenicity in the vein graft. METHODS AND RESULTS: Autologous rabbit jugular vein grafts were incubated ex vivo in a solution of adenovirus vectors containing CNP gene (Ad.CNP) or Escherichia coli lac Z gene (Ad.LacZ) and then interposed in the carotid artery. Reendothelialization, mural thrombi formation, and intima/media ratio were evaluated on the 14th and 28th postoperative days. More reendothelialization was seen in Ad.CNP-infected grafts than in Ad.LacZ-infected grafts both at 14 days (0.81+/-0.05 versus 0.30+/-0.14, P<0.01) and at 28 days (0.96+/-0.01 versus 0.45+/-0.08, P<0.001). The mural thrombus area was smaller in Ad.CNP-infected grafts than in Ad.LacZ-infected grafts. Neointimal thickening was significantly suppressed in the Ad.CNP group. The in vitro wound assay with human coronary artery endothelial cells revealed significant potentiation of the wound repair process by CNP and atrial natriuretic peptide administration. CONCLUSIONS: Infected Ad.CNP accelerated reendothelialization and suppressed thrombosis and neointimal hyperplasia. The method may potentially prevent vein graft disease in patients undergoing coronary artery revascularization. PMID- 11940537 TI - Identification of atherosclerotic plaque components with intravascular ultrasound elastography in vivo: a Yucatan pig study. AB - BACKGROUND: Intravascular ultrasound elastography assesses the local strain of the atherosclerotic vessel wall. In the present study, the potential to identify different plaque components in vivo was investigated. METHODS AND RESULTS: Atherosclerotic external iliac and femoral arteries (n=24) of 6 Yucatan pigs were investigated. Before termination, elastographic data were acquired with a 20-MHz Visions catheter. Two frames acquired at end-diastole with a pressure differential of approximately 4 mm Hg were acquired to obtain the elastograms. Before dissection, x-ray was used to identify the arterial segments that had been investigated by ultrasound. Specimens were stained for collagen, fat, and macrophages. Plaques were classified as absent, early fibrous lesion, early fatty lesion, or advanced fibrous plaque. The average strains in the plaque-free arterial wall (0.21%) and the early (0.24%) and advanced fibrous plaques (0.22%) were similar. Higher average strain values were observed in fatty lesions (0.46%) compared with fibrous plaques (P=0.007). After correction for confounding by lipid content, no additional differences in average strain were found between plaques with and without macrophages (P=0.966). Receiver operating characteristic analysis revealed a sensitivity and a specificity of 100% and 80%, respectively, to identify fatty plaques. The presence of a high-strain spot (strain >1%) has 92% sensitivity and 92% specificity to identify macrophages. CONCLUSIONS: To the best of our knowledge, this is the first time that intravascular ultrasound elastography has been validated in vivo. Fatty plaques have an increased mean strain value. High-strain spots are associated with the presence of macrophages. PMID- 11940538 TI - Optical imaging of cardiac reporter gene expression in living rats. AB - BACKGROUND: Studies of cardiac gene transfer rely on postmortem analysis using histologic staining or enzyme assays. Noninvasive imaging of the temporal and spatial characteristics of cardiac gene expression in the same subject offers significant advantages. METHODS AND RESULTS: Rats underwent direct myocardial injection via left thoracotomy with adenovirus-expressing firefly luciferase (Ad CMV-Fluc; n=30). The reporter substrate D-luciferin was injected intraperitoneally. Serial images were acquired by use of a cooled charged couple detector (CCD) camera. Results are expressed as relative light unit per minute (RLU/min). Rats transduced with 1x10(9) plaque-forming units show decremental cardiac luciferase activity over time: 152 070+/-21 170 (day 2), 195 806+/-62 630 (day 5), 7250+/-2941 (day 8), and 2040+/-971 RLU/min (day 14). To assess the detection sensitivity, serially diluted titers of Ad-CMV-Fluc were injected: 1x10(9) (195 393+/-14 896), 1x10(8) (33 777+/-18 179), 1x10(7) (417+/-91), 1x10(6) (185+/-64), 1x10(5) (53+/-1), and control (54+/-1) (P<0.05 for 1x10(9), 1x10(8), and 1x10(7) plaque-forming units versus control adenovirus-expressing mutant thymidine kinase [Ad-CMV-HSV1-sr39tk]; n=3). Finally, rats were euthanized, and in vitro luciferase activity correlated with in vivo CCD signals (r2=0.92). CONCLUSIONS: This study demonstrates for the first time the feasibility of imaging the location, magnitude, and time course of cardiac reporter gene expression in living rats. Cardiac gene therapy studies could be aided with wider application of this approach. PMID- 11940539 TI - Cardiovascular aging is associated with vitamin E increase. AB - BACKGROUND: Aging is an independent risk factor for the development of cardiovascular disease. Therefore, therapies to delay vascular aging may have enormous medical consequences. In this context, vitamin E is of particular interest, mainly because of its antioxidative properties. METHODS AND RESULTS: In 3-year-old rats, which are not susceptible to atherosclerosis, vitamin E levels, as measured by reversed-phase high-performance liquid chromatography, were markedly increased both in plasma and in major organs (P<0.01 to P<0.0001). The highest increase (at least 70-fold) was found in the aortic wall. CONCLUSIONS: This unexpected accumulation of vitamin E appears to be a compensatory mechanism that attempts to counterbalance age-associated oxidative stress and that may represent a self-regulatory protective adaptation. PMID- 11940540 TI - Direct evidence for tumor necrosis factor-alpha signaling in arteriogenesis. AB - BACKGROUND: Arteriogenesis serves as an efficient mechanism for flow restoration after arterial occlusion. This process is associated with inflammatory mediators such as tumor necrosis factor-alpha (TNF-alpha), although their role in arteriogenesis remains unclear. We hypothesized that arteriogenesis is reduced in mice lacking functional TNF-alpha or p55 receptor. To test this hypothesis, we developed a novel microsphere-based murine model of hindlimb perfusion measurement. METHODS AND RESULTS: Unilateral femoral arteries of nude (n=9), TNF alpha(-/-) (n=9), TNF-alpha receptor p55(-/-) (n=8), and p75(-/-) (n=8) mice as well as their appropriate genetic background controls were occluded. The nude mice underwent laser Doppler hindlimb flux measurements preoperatively, postoperatively, and after 7 days. Seven days after ligation, all animals underwent tissue perfusion determinations using fluorescent microspheres. Laser Doppler findings confirmed acute decrease in flux with falsely normal values after 1 week. Microsphere results from control mice showed perfusion restoration to values approximately 50% of normal within 7 days. TNF-alpha(-/-) mice demonstrated a significant reduction (45.1%) in collateral artery perfusion compared with controls (TNF-alpha(-/-) 22.4+/-5.1% versus B6x129 49.7+/-9.3%; P<0.01). p55(-/-) mice exhibited an almost identical 45.8% reduction in collateral artery formation (p55(-/-) 28.3+/-4.3% versus C57BL/6J 61.8+/-9.1%; P<0.01), whereas p75(-/-) mice were equivalent to controls (p75(-/-) 54.5+/-5.5%; P=0.13). CONCLUSIONS: Microsphere techniques in mice offer a tool for the molecular dissection of arteriogenesis mechanisms. These results suggest that TNF alpha positively modulates arteriogenesis probably via signaling through its p55 receptor. PMID- 11940541 TI - Enoxaparin as adjunctive antithrombin therapy for ST-elevation myocardial infarction: results of the ENTIRE-Thrombolysis in Myocardial Infarction (TIMI) 23 Trial. AB - BACKGROUND: ENTIRE-TIMI 23 evaluated enoxaparin with full-dose tenecteplase (TNK) and half-dose TNK plus abciximab. METHODS AND RESULTS: Patients (n=483) with ST elevation MI presenting <6 hours from symptom onset were randomized to full-dose TNK and either unfractionated heparin (UFH) (bolus 60 U/kg; infusion 12 U/kg per hour) or enoxaparin (1.0 mg/kg subcutaneously every 12 hours+/-initial 30 mg intravenous bolus), or half-dose TNK plus abciximab and either UFH (bolus 40 U/kg; infusion 7 U/kg per hour) or enoxaparin (0.3 to 0.75 mg/kg subcutaneously every 12 hours+/-initial intravenous bolus of 30 mg). With full-dose TNK and UFH, the rate of TIMI 3 flow at 60 minutes was 52% and was 48% to 51% with enoxaparin. Using combination therapy, the rate of TIMI 3 flow was 48% with UFH and 47% to 58% with enoxaparin. The rate of TIMI 3 flow among all UFH patients was 50% and was 51% among enoxaparin patients. Through 30 days, death/recurrent MI occurred in the full-dose TNK group in 15.9% of patients with UFH and 4.4% with enoxaparin (P=0.005). In the combination therapy group, the rates were 6.5% with UFH and 5.5% with enoxaparin. The rate of major hemorrhage with full-dose TNK was 2.4% with UFH and 1.9% with enoxaparin; with combination therapy, it was 5.2% using UFH and 8.5% with enoxaparin. CONCLUSIONS: Enoxaparin is associated with similar TIMI 3 flow rates as UFH at an early time point while exhibiting advantages over UFH with respect to ischemic events through 30 days. These findings with enoxaparin are achieved with a similar risk of major hemorrhage. PMID- 11940542 TI - Aspirin-resistant thromboxane biosynthesis and the risk of myocardial infarction, stroke, or cardiovascular death in patients at high risk for cardiovascular events. AB - BACKGROUND: We studied whether aspirin resistance, defined as failure of suppression of thromboxane generation, increases the risk of cardiovascular events in a high-risk population. METHODS AND RESULTS: Baseline urine samples were obtained from 5529 Canadian patients enrolled in the Heart Outcomes Prevention Evaluation (HOPE) Study. Using a nested case-control design, we measured urinary 11-dehydro thromboxane B2 levels, a marker of in vivo thromboxane generation, in 488 cases treated with aspirin who had myocardial infarction, stroke, or cardiovascular death during 5 years of follow-up and in 488 sex- and age-matched control subjects also receiving aspirin who did not have an event. After adjustment for baseline differences, the odds for the composite outcome of myocardial infarction, stroke, or cardiovascular death increased with each increasing quartile of 11-dehydro thromboxane B2, with patients in the upper quartile having a 1.8-times-higher risk than those in the lower quartile (OR, 1.8; 95% CI, 1.2 to 2.7; P=0.009). Those in the upper quartile had a 2-times higher risk of myocardial infarction (OR, 2.0; 95% CI, 1.2 to 3.4; P=0.006) and a 3.5-times-higher risk of cardiovascular death (OR, 3.5; 95% CI, 1.7 to 7.4; P<0.001) than those in the lower quartile. CONCLUSIONS: In aspirin-treated patients, urinary concentrations of 11-dehydro thromboxane B2 predict the future risk of myocardial infarction or cardiovascular death. These findings raise the possibility that elevated urinary 11-dehydro thromboxane B2 levels identify patients who are relatively resistant to aspirin and who may benefit from additional antiplatelet therapies or treatments that more effectively block in vivo thromboxane production or activity. PMID- 11940543 TI - Mechanisms of increased vascular superoxide production in human diabetes mellitus: role of NAD(P)H oxidase and endothelial nitric oxide synthase. AB - BACKGROUND: Increased superoxide production contributes to reduced vascular nitric oxide (NO) bioactivity and endothelial dysfunction in experimental models of diabetes. We characterized the sources and mechanisms underlying vascular superoxide production in human blood vessels from diabetic patients with coronary artery disease compared with nondiabetic patients. METHODS AND RESULTS: Vascular superoxide production was quantified in both saphenous veins and internal mammary arteries from 45 diabetic and 45 matched nondiabetic patients undergoing coronary artery bypass surgery. NAD(P)H-dependent oxidases were important sources of vascular superoxide in both diabetic and nondiabetic patients, but both the activity of this enzyme system and the levels of NAD(P)H oxidase protein subunits (p22phox, p67phox, and p47phox) were significantly increased in diabetic veins and arteries. In nondiabetic vessels, endothelial NO synthase produced NO that scavenged superoxide. However, in diabetic vessels, the endothelium was an additional net source of superoxide production because of dysfunctional endothelial NO synthase that was corrected by intracellular tetrahydrobiopterin supplementation. Furthermore, increased superoxide production in diabetes was abrogated by the protein kinase C inhibitor chelerythrine. CONCLUSIONS: These observations suggest important roles for NAD(P)H oxidases, endothelial NO synthase uncoupling, and protein kinase C signaling in mediating increased vascular superoxide production and endothelial dysfunction in human diabetes mellitus. PMID- 11940544 TI - Pulmonary hypertension and risk of death in cardiomyopathy: patients with myocarditis are at higher risk. AB - BACKGROUND: Pulmonary hypertension is a clinically useful predictor of death in patients with heart failure. Whether pulmonary hypertension has the same prognostic value among specific underlying causes of cardiomyopathy is unknown. Using a diverse cohort of cardiomyopathy patients, we tested the hypotheses that (1) elevated mean pulmonary arterial pressure is the most important hemodynamic predictor of death and (2) the prognostic value of mean pulmonary pressure varies among different cardiomyopathies. METHODS AND RESULTS: Patients (n=1134) with new cardiomyopathy were prospectively assigned a specific diagnosis on the basis of clinical evaluation and endomyocardial biopsy. All patients underwent right heart catheterization at baseline and were followed for an average of 4.4 years. In multivariate Cox models that allowed for nonlinear relations between hemodynamics and death, mean systemic pressure (mSP) and mean pulmonary arterial pressure (mPA) emerged as the most important hemodynamic predictors of death. Moreover, there was a statistically significant positive interaction between mPA and the diagnosis of myocarditis. For each 5-mm Hg increase in baseline mSP, mortality rates decreased with relative hazard (RH) of 0.89 (0.86 to 0.92). For a 5-mm Hg increase in baseline mPA, mortality rates increased in patients who did not carry the diagnosis of myocarditis with RH 1.23 (1.17 to 1.29); among patients with myocarditis, mortality rates increased substantially with RH of 1.85 (1.50 to 2.29; P<0.001 for interaction). CONCLUSIONS: Baseline mPA is particularly important for stratifying risk in myocarditis. These findings suggest that secondary pulmonary hypertension may have different biological features in myocarditis and that patients with pulmonary hypertension and myocarditis should be targeted for aggressive medical therapy. PMID- 11940545 TI - Paradoxical pressor effects of beta-blockers in standing elderly patients with mild hypertension: a beneficial side effect. AB - BACKGROUND: Baroreflex sensitivity declines with age, creating a fall in systolic blood pressure and pulse pressure when standing. If, in addition, blood pressure is reduced as a result of antihypertensive medication, compensatory mechanisms may be inadequate and orthostatic problems may occur. This may be less true in patients on beta-blockers. beta-blockers cause pressor effects in standing patients with autonomic neuropathy, but their effects on standing pulse pressures in elderly subjects with mild hypertension have not been systematically studied. METHODS AND RESULTS: We studied 3741 patients with mild hypertension for 6 months who were being treated with the beta-blocker nebivolol 5 mg daily. Blood pressures were measured after 10 minutes in the supine position and after 1 minute in the standing position. Overall, systolic and diastolic blood pressures rose slightly while standing, whereas pulse pressures remained unchanged. When previously untreated patients (n=2085) >60 and <60 years of age were assessed separately, supine pulse pressures were consistently higher in the elderly group compared with those of the younger subjects by 6 to 11 mm Hg (P<0.001 to 0.0001). However, while standing, pulse pressures rose in the younger subjects, whereas they tended to fall in the elderly group. After 6 months of beta-blockade, this pattern was unchanged in the younger subjects but reversed into significant rise of pulse pressures in the elderly group by 4 (SD 1) mm Hg (P<0.001). In the patients previously treated with other classes of antihypertensive drugs (n=712), the effects were essentially the same. CONCLUSIONS: In elderly patients with mild hypertension, a depressor trend of pulse pressure while standing can be turned into a significant pressor response by treatment with a beta-blocker. PMID- 11940546 TI - Primary pulmonary hypertension is associated with reduced pulmonary vascular expression of type II bone morphogenetic protein receptor. AB - BACKGROUND: Mutations in the type II receptor for bone morphogenetic protein (BMPR-II), a receptor member of the transforming growth factor-beta (TGF-beta) superfamily, underlie many familial and sporadic cases of primary pulmonary hypertension (PPH). METHODS AND RESULTS: Because the sites of expression of BMPR II in the normal and hypertensive lung are unknown, we studied the cellular localization of BMPR-II and the related type I and II receptors for TGF-beta by immunohistochemistry in lung sections from patients undergoing heart-lung transplantation for PPH (n=11, including 3 familial cases) or secondary pulmonary hypertension (n=6) and from unused donor lungs (n=4). In situ hybridization was performed for BMPR-II mRNA. Patients were screened for the presence of mutations in BMPR2. In normal lungs, BMPR-II expression was prominent on vascular endothelium, with minimal expression in airway and arterial smooth muscle. In pulmonary hypertension cases, the intensity of BMPR-II immunostaining varied between lesions but involved endothelial and myofibroblast components. Image analysis confirmed that expression of BMPR-II was markedly reduced in the peripheral lung of PPH patients, especially in those harboring heterozygous BMPR2 mutations. A less marked reduction was also observed in patients with secondary pulmonary hypertension. In contrast, there was no difference in level of staining for TGF-betaRII or the endothelial marker CD31. CONCLUSIONS: The cellular localization of BMPR-II is consistent with a role in the formation of pulmonary vascular lesions in PPH, and reduced BMPR-II expression may contribute to the process of vascular obliteration in severe pulmonary hypertension. PMID- 11940547 TI - Markers of increased risk of intracerebral hemorrhage after intravenous recombinant tissue plasminogen activator therapy for acute ischemic stroke in clinical practice: the Multicenter rt-PA Stroke Survey. AB - BACKGROUND: Intravenous recombinant tissue plasminogen activator (rtPA) is an effective therapy for acute ischemic stroke, but it is associated with risk of intracerebral hemorrhage (ICH). Our aim was to identify, in a large cohort of patients, readily available baseline factors that are associated with thrombolysis-related ICH. METHODS AND RESULTS: In a multicenter retrospective and prospective investigation of individual data from 1205 patients treated in routine clinical practice with intravenous rtPA within 3 hours of stroke symptom onset, 72 patients (6%) developed symptomatic ICH and 86 additional patients (7%) had asymptomatic ICH identified on a routine follow-up CT. In analyses based on clinical variables alone, the main attributes associated with ICH were a history of diabetes mellitus and cardiac disease, increasing stroke severity, advancing age, use of antiplatelet agents other than aspirin before stroke onset, and elevated pretreatment mean blood pressure. In additional analyses that incorporated baseline CT and laboratory findings (in a subset of patients), the main associations were early ischemic CT changes, in particular if exceeding one third of middle cerebral artery territory; increasing stroke severity; diabetes mellitus or elevated serum glucose; and lower platelet counts. Final independent attributes associated with parenchymatous hematoma, defined by purely radiologically based criteria, were similar to those of symptomatic ICH. CONCLUSIONS: Readily available factors can identify acute ischemic stroke patients at high and low risk for rtPA-related ICH. These factors require confirmation in a prospective cohort before clinical implementation. PMID- 11940548 TI - Endothelial healing in vein grafts: proliferative burst unimpaired by genetic therapy of neointimal disease. AB - BACKGROUND: Although inhibition of neointimal hyperplasia by cell cycle gene blockade therapy results in improved endothelial cell function in experimental vein grafts, little is known either about endothelial healing immediately after vein grafting or about the effect of this therapy on the healing process. METHODS AND RESULTS: Scanning electron microscopy demonstrated an immediate decrease in vein graft endothelial cell density associated with vein graft wall stretch, followed by a return to baseline by postoperative day 3. En face detection of bromodeoxyuridine incorporation confirmed a rapid endothelial proliferation by 48 hours. Despite inhibition of underlying vascular smooth muscle cell proliferation, E2F decoy oligonucleotide did not inhibit either endothelial bromodeoxyuridine incorporation or the return to baseline cell density. This differential response to E2F decoy was also observed in human umbilical vein endothelial cell culture, which resisted the E2F decoy inhibition of cell growth that was observed in human umbilical artery smooth muscle cells, despite evidence for nuclear localized delivery of the oligonucleotide into both cell types. Furthermore, the reduction of E2F binding activity seen in a nuclear gel shift assay of cultured smooth muscle cells was not observed in endothelial cells. CONCLUSIONS: These results suggest a burst of graft endothelial cell proliferation that allows a rapid restoration of cell density in the monolayer. Additionally, there is a selective effect of E2F decoy gene therapy on target smooth muscle cells with sparing of this endothelial healing. PMID- 11940549 TI - Validation of myocardial acceleration during isovolumic contraction as a novel noninvasive index of right ventricular contractility: comparison with ventricular pressure-volume relations in an animal model. AB - BACKGROUND: We have demonstrated that myocardial acceleration during isovolumic contraction (IVA) is a sensitive index of left ventricular contractile function. In this study, we assessed the utility of IVA to measure right ventricular (RV) contractile function. METHODS AND RESULTS: We examined 8 pigs by using tissue Doppler imaging of the RV free wall and simultaneous measurements of intraventricular pressure, volume, maximal elastance (e(max)), preload recruitable stroke work, and dP/dt(max) by conductance catheterization. Animals were paced in the right atrium at a rate of 130 beats per minute (bpm). IVA was compared with elastance during contractility modulation by esmolol and dobutamine and during preload reduction and afterload increase by transient balloon occlusion of the inferior vena cava and pulmonary artery, respectively. Data were also obtained during incremental atrial pacing from 110 to 210 bpm. Esmolol led to a decrease in IVA and dP/dt(max). During dobutamine infusion, IVA, dP/dt(max), preload recruitable stroke work, and e(max) all increased significantly. During preload reduction and afterload increase, IVA remained constant up to a reduction of RV volume by 54% and an RV systolic pressure increase of 58%. Pacing up to a rate of 190 bpm led to a stepwise increase in IVA and dP/dt(max), with a subsequent fall at a pacing rate of 210 bpm. CONCLUSIONS: IVA is a measurement of RV contractile function that is unaffected by preload and afterload changes in a physiological range and is able to measure the force-frequency relation. This novel index may be ideally suited to the assessment of acute changes of RV function in clinical studies. PMID- 11940550 TI - Cardiovascular influences of alpha1b-adrenergic receptor defect in mice. AB - BACKGROUND: The alpha1-adrenergic receptors (alpha1-ARs) play a key role in cardiovascular homeostasis. However, the functional role of alpha1-AR subtypes in vivo is still unclear. The aim of this study was to evaluate the cardiovascular influences of alpha1b-AR. METHODS AND RESULTS: In transgenic mice lacking alpha1 AR (KO) and their wild-type controls (WT), we evaluated blood pressure profile and cardiovascular remodeling induced by the chronic administration (18 days via osmotic pumps) of norepinephrine, angiotensin II, and subpressor doses of phenylephrine. Our results indicate that norepinephrine induced an increase in blood pressure levels only in WT mice. In contrast, the hypertensive state induced by angiotensin II was comparable between WT and KO mice. Phenylephrine did not modify blood pressure levels in either WT or KO mice. The cardiac hypertrophy and eutrophic vascular remodeling evoked by norepinephrine was observed only in WT mice, and this effect was independent of the hypertensive state because it was similar to that observed during subpressor phenylephrine infusion. Finally, the cardiac hypertrophy induced by thoracic aortic constriction was comparable between WT and KO mice. CONCLUSIONS: Our data demonstrate that the lack of alpha1b-AR protects from the chronic increase of arterial blood pressure induced by norepinephrine and concomitantly prevents cardiovascular remodeling evoked by adrenergic activation independently of blood pressure levels. PMID- 11940551 TI - Vessel imaging by interferometric phase-contrast X-ray technique. AB - BACKGROUND: Phase-contrast x-ray imaging using an x-ray interferometer has great potential to reveal the structures inside soft tissues, because the sensitivity of this method to hydrogen, carbon, nitrogen, and oxygen is approximately 1000 times higher than that of the absorption-contrast x-ray method. Imaging of vessels is very important to understand the vascular distribution of organs and tumors, so the possibility of selective angiography based on phase contrast is examined with a physiological material composed of low-atomic-number elements. METHODS AND RESULTS: Phase-contrast x-ray imaging was performed with a synchrotron x-ray source. Differences in refractive index, ddelta, of physiological saline, lactated Ringer's solution, 5% glucose, artificial blood such as pyridoxylated hemoglobin-polyoxyethylene conjugate, and perfluorotributylamine were measured. Because the ddelta of physiological saline has highest contrast, it was used for the phase-contrast x-ray imaging of vessel, and this was compared with absorption-contrast x-ray images. Vessels >0.03 mm in diameter of excised liver from rats and a rabbit were revealed clearly in phase contrast x-ray imaging, whereas the vessel could not be revealed at all by the absorption-contrast x-ray image. Absorption-contrast x-ray images with iodine microspheres depicted only portal veins >0.1 mm in diameter with nearly the same x-ray dose as the present phase-contrast x-ray imaging. CONCLUSIONS: Phase contrast x-ray imaging explored clear depiction of the vessels using physiological saline with small doses of x-rays. PMID- 11940552 TI - Imaging of oxygen transfer among microvessels of rat cremaster muscle. AB - BACKGROUND: The proximity of capillaries, arterioles, and venules provides complex spatial relationships that lead to oxygen transfer among microvessels. Although a conceptual image of complex oxygen transfer among microvessels has been hypothesized, in vivo mapping of oxygen saturation (SO2) levels in microvessels had never been performed. METHODS AND RESULTS: The oxygen profile of the arterioles and venules of the rat cremaster muscle during normoxia and hypoxia was visualized by preparing pseudo-color images of SO2 levels based on microspectrophotometry data obtained by using 3 different optical filters and a cooled CCD camera. The SO2 images showed lower SO2 levels in arterioles close to their walls, and the SO2 levels in the paired venules showed higher SO2 levels close to the arterioles. There were capillaries that crossed the microvessels whose SO2 levels changed as they crossed the microvessels. The SO2 levels were lower close to the vessel wall than in the centerline level of the microvessels, and the highest SO2 levels in venules paralleling arterioles were skewed toward the arterial side. The SO2 images showed that the SO2 level in arterioles decreased after crossing venules, whereas the SO2 level in venules increased after crossing arterioles. CONCLUSIONS: Visualization of intravascular SO2 levels suggested that oxygen is transferred between paired microvessels and between crossing microvessels in rat cremaster muscle. The possibility that oxygen is transported from some arterioles to venules and tissue through adjacent capillaries is proposed. PMID- 11940553 TI - Rebuilding a damaged heart: long-term survival of transplanted neonatal rat cardiomyocytes after myocardial infarction and effect on cardiac function. AB - BACKGROUND: The long-term effects of cardiac cell transplantation on cardiac function are unknown. Therefore, we tested the survival and functional impact of rat neonatal cardiac myocytes up to 6 months after transplantation into infarcted hearts. METHODS AND RESULTS: Cardiomyocytes from male neonatal Fischer 344 rats (1 to 2 days, 3 to 5x10(6)) or medium was injected into the infarcts of adult syngeneic female animals 1 week after left coronary artery ligation. Six months later, implanted cardiomyocytes were still present by quantitative TaqMan polymerase chain reaction and histology. In all treated hearts, discrete lumps of cells were present within the infarct scar, which was not observed in media injected hearts typified by a transmural infarct scar. Infarct thickness was greater in treated animals versus control animals (909+/-97 versus 619+/-43 microm, P<0.02), whereas infarct size and left ventricular volumes were similar. By biplane angiography, left ventricular ejection fractions at 6 months were greater (0.36+/-0.03 versus 0.25+/-0.02, P<0.01) and significantly less infarct zone dyskinesis was seen (0.30+/-0.08 versus 0.55+/-0.07, P=0.035, lateral projection) in treated animals versus control animals. CONCLUSIONS: Grafted neonatal cardiomyocytes were present in infarcts 6 months after transplantation; they thickened the wall of the left ventricle and were associated with enhanced ejection fraction and reduced paradoxical systolic bulging of the infarct. Therefore, neonatal cardiac cell transplants exhibit long-term survival in a myocardial infarct model and contribute to long-term improved cardiac function. These results suggest that a damaged heart can be rebuilt. PMID- 11940554 TI - Adaptation and maladaptation of the heart in diabetes: Part I: general concepts. PMID- 11940555 TI - Images in cardiovascular medicine. False left ventricular aneurysm documented by magnetic resonance imaging. PMID- 11940557 TI - Images in cardiovascular medicine. Persistent left superior vena cava demonstrated with multislice spiral computed tomography. PMID- 11940556 TI - Secondary prevention of coronary heart disease in the elderly (with emphasis on patients > or =75 years of age): an American Heart Association scientific statement from the Council on Clinical Cardiology Subcommittee on Exercise, Cardiac Rehabilitation, and Prevention. PMID- 11940558 TI - Demonstration of the exact anatomic tachycardia circuit in the fast-slow form of atrioventricular nodal reentrant tachycardia. PMID- 11940559 TI - A pill for every ill. PMID- 11940560 TI - Chronic immune stimulation may link ischemic heart disease with depression. PMID- 11940561 TI - Use of selective serotonin reuptake inhibitors and myocardial infarction. PMID- 11940562 TI - Safety and efficacy of unprotected left main coronary artery stenting. PMID- 11940564 TI - Identifying the stroke victim at risk for hemorrhage during thrombolysis. PMID- 11940565 TI - Role of calcium and membrane organization on phospholipase D localization and activity. Competition between a soluble and insoluble substrate. AB - The phospholipase D (PLD) from Streptomyces chromofuscus is a soluble enzyme known to be activated by the phosphatidic acid-calcium complexes. PLD-catalyzed hydrolysis of phospholipids in aqueous medium leads to the formation of phosphatidic acid (PA). Previous studies concluded on an allosteric activation of PLD by the PA-calcium complexes. In this work, the role of PA and calcium was investigated in terms of membrane structure and dynamics. The role of calcium in PLD partitioning between the soluble phase and the water-lipid interface was tested. The monomolecular film technique was used to measure both membrane dynamics and PLD activity. These experiments provided information on PLD activity at a water-lipid interface. Moreover, the ability of PA to enhance PLD activity toward phosphatidylcholine was correlated to the physical properties of PA itself, affecting the rheology of the membrane. The effect of calcium was investigated on PLD binding to lipids and on the catalytic process by competition experiments between a soluble and a vesicular substrate. These experiments confirmed the absolute PLD requirement for calcium and pointed out the importance of calcium for PLD catalytic process and for the enzyme location at the water lipid interface. PMID- 11940566 TI - The role of topoisomerase II in the excision of DNA loop domains during apoptosis. AB - Disintegration of nuclear DNA into high molecular weight (HMW) and oligonucleosomal DNA fragments represents two major periodicities of DNA fragmentation during apoptosis. These are thought to originate from the excision of DNA loop domains and from the cleavage of nuclear DNA at the internucleosomal positions, respectively. In this report, we demonstrate that different apoptotic insults induced apoptosis in NB-2a neuroblastoma cells that was invariably accompanied by the formation of HMW DNA fragments of about 50-100 kb but proceeded either with or without oligonucleosomal DNA cleavage, depending on the type of apoptotic inducer. We demonstrate that differences in the pattern of DNA fragmentation were reproducible in a cell-free apoptotic system and develop conditions that allow in vitro separation of the HMW and oligonucleosomal DNA fragmentation activities. In contrast to apoptosis associated with oligonucleosomal DNA fragmentation, the HMW DNA cleavage in apoptotic cells was accompanied by down-regulation of caspase-activated DNase (CAD) and was not affected by z-VAD-fmk, suggesting that the caspase/CAD pathway is not involved in the excision of DNA loop domains. We further demonstrate that nonapoptotic NB-2a cells contain a constitutively present nuclease activity located in the nuclear matrix fraction that possessed the properties of topoisomerase (topo) II and was capable of reproducing the pattern of HMW DNA cleavage that occurred in apoptotic cells. We demonstrate that the early stages of apoptosis induced by different stimuli were accompanied by activation of topo II-mediated HMW DNA cleavage that was reversible after removal of apoptotic inducers, and we present evidence of the involvement of topo II in the formation of HMW DNA fragments at the advanced stages of apoptosis. The results suggest that topo II is involved in caspase independent excision of DNA loop domains during apoptosis, and this represents an alternative pathway of apoptotic DNA disintegration from CAD-driven caspase dependent oligonucleosomal DNA cleavage. PMID- 11940567 TI - Phosphorylation of STAT-3 in response to basic fibroblast growth factor occurs through a mechanism involving platelet-activating factor, JAK-2, and Src in human umbilical vein endothelial cells. Evidence for a dual kinase mechanism. AB - Platelet-activating factor (PAF) is a potent proinflammatory phospholipid with multiple pathological and physiological effects. We have shown that basic fibroblast growth factor (bFGF) supplementation induces rapid proliferation of human umbilical vein endothelial cells (HUVEC), which is reduced upon removal of bFGF or by bFGF immunoneutralization. The PAF receptor antagonist LAU-8080 inhibited bFGF-stimulated HUVEC proliferation, indicating the involvement of PAF in the bFGF-mediated signaling of HUVEC. Although FGF receptor phosphorylation was not affected by LAU-8080, the bFGF-mediated prolonged phosphorylation, and activation of Erk-1 and -2 were attenuated. Phosphorylation of STAT-3 was observed in the presence of PAF or bFGF, which was attenuated by PAFR antagonists. PAF-induced STAT-3 phosphorylation observed in HUVEC pretreated with either Src inhibitor PP1 or JAK-2 inhibitor AG-490 indicated (i) immediate (1 min) phosphorylation of STAT-3 is dependent on Src, (ii) JAK-2-dependent STAT-3 phosphorylation occurs after the delayed (30 min) PAF exposure, and (iii) prolonged (60 min) STAT-3 phosphorylation may be either through Src and/or JAK-2. Attenuation of the STAT-3 phosphorylation by the PAFR antagonists indicated signaling through the PAF receptor. Taken together, these findings suggest the production of PAF is important for bFGF-mediated signaling and that a dual kinase mechanism is involved in the PAF-mediated signal transduction cascade. PMID- 11940568 TI - Post-transcriptional regulation of luteinizing hormone receptor mRNA in the ovary by a novel mRNA-binding protein. AB - Luteinizing hormone (LH) receptor mRNA is post-transcriptionally regulated. An ovarian cytosolic LH receptor mRNA-binding protein (LRBP) identified in our laboratory binds to a polypyrimidine-rich bipartite sequence in the coding region of LH receptor mRNA. The present studies show a role for LRBP in the regulation of LH receptor mRNA. We demonstrated that increased LH receptor mRNA degradation occurs during hormone-induced LH receptor down-regulation. Furthermore, increased degradation of LH receptor mRNA was seen when partially purified LRBP was included in an in vitro mRNA decay reaction. The LH receptor mRNA binding activity of LRBP measured by RNA electrophoretic mobility shift analysis showed an inverse relationship to LH receptor mRNA levels during different physiological states. These results suggest that LRBP is a physiological regulator of LHR mRNA expression in the ovary and provides a novel mechanism for the regulation of LH receptor expression in the ovary. PMID- 11940569 TI - Purification, reconstitution, and steady-state kinetics of the trans-membrane 17 beta-hydroxysteroid dehydrogenase 2. AB - Human membrane 17 beta-hydroxysteroid dehydrogenase 2 is an enzyme essential in the conversion of the highly active 17beta-hydroxysteroids into their inactive keto forms in a variety of tissues. 17 beta-hydroxysteroid dehydrogenase 2 with 6 consecutive histidines at its N terminus was expressed in Sf9 insect cells. This recombinant protein retained its biological activity and facilitated the enzyme purification and provided the most suitable form in our studies. Dodecyl-beta-D maltoside was found to be the best detergent for the solubilization, purification, and reconstitution of this enzyme. The overexpressed integral membrane protein was purified with a high catalytic activity and a purity of more than 90% by nickel-chelated chromatography. For reconstitution, the purified protein was incorporated into dodecyl-beta-D-maltoside-destabilized liposomes prepared from l-alpha-phosphatidylcholine. The detergent was removed by adsorption onto polystyrene beads. The reconstituted enzyme had much higher stability and catalytic activity (2.6 micromol/min/mg of enzyme protein with estradiol) than the detergent-solubilized and purified protein (0.9 micromol/min/mg of enzyme protein with estradiol). The purified and reconstituted protein (with a 2-kDa His tag) was proved to be a homodimer, and its functional molecular mass was calculated to be 90.4 +/- 1.2 kDa based on glycerol gradient analytical ultracentrifugation and chemical cross-linking study. The kinetic studies demonstrated that 17 beta-hydroxysteroid dehydrogenase 2 was an NAD preferring dehydrogenase with the K(m) of NAD being 110 +/- 10 microM and that of NADP 9600 +/- 100 microM using estradiol as substrate. The kinetic constants using estradiol, testosterone, dihydrotestosterone, and 20 alpha dihydroprogesterone as substrates were also determined. PMID- 11940570 TI - Lipopolysaccharide-mediated reactive oxygen species and signal transduction in the regulation of interleukin-1 gene expression. AB - Lipopolysaccharide (LPS) stimulates macrophages to release inflammatory cytokines, interleukin-1 beta (IL-1), and tumor necrosis factor (TNF). LPS induced TNF suppresses scavenger receptor functions in macrophages (van Lenten, B. J., and Fogelman, A. M. (1992) J. Immunol. 148, 112-116), which is regulated by TNF-mediated protein kinases (Hsu, H. Y., and Twu, Y. C. (2000) J. Biol. Chem. 275, 41035-41048). To examine the molecular mechanism for LPS induction of IL-1 in macrophages, we demonstrated that LPS quickly stimulated reactive oxygen species (ROS), and 3 h later induced prointerleukin-1 beta (pro-IL-1, precursor of IL-1) production and IL-1 secretion. LPS stimulated pro-IL-1 message/protein between 3 and 10 h; however, there was a 40% reduction of pro-IL-1 in preincubation of the antioxidant, N-acetylcysteine (NAC). Moreover, NAC moderated LPS-induced IL-1 secretion partially via interleukin 1-converting enzyme. The maximal activity of LPS-induced ERK, JNK, and p38 was 12- (30 min), 5- (30 min), and 16-fold (15 min), respectively. In contrast, NAC reduced ERK activity to 60% and decreased p38 activity to the basal level, but JNK activity was induced 2 fold. Furthermore, the pharmacological antagonists LY294002, SB203580, curcumin, calphostin C, and PD98059 revealed the diverse roles of LPS-mediated protein kinases in pro-IL-1. On the other hand, NAC and diphenyleneiodonium chloride partially inhibited LPS-induced Rac activity and protein-tyrosine kinase (PTK), indicating that LPS-mediated ROS and NADPH oxidase correspond to Rac activation and IL-1 expression. Our findings establish for the first time that LPS-mediated PTK/phosphatidylinositol 3-kinase/Rac/p38 pathways play a more important role than pathways of PTK/PKC/MEK/ERK and of PTK/phosphatidylinositol 3-kinase/Rac/JNK in the regulation of pro-IL-1/IL-1. The findings also further elucidate the critical role of LPS-mediated ROS in signal transduction pathways. Our results suggest that understanding LPS-transduced signals in IL-1 induction upon the antibacterial action of macrophages should provide a therapeutic strategy for aberrant inflammatory responses leading to severe cellular injury or concurrent multiorgan septic damage. PMID- 11940571 TI - The role of zinc ions in reverse transport mediated by monoamine transporters. AB - The human dopamine transporter (hDAT) contains an endogenous high affinity Zn2+ binding site with three coordinating residues on its extracellular face (His193, His375, and Glu396). Upon binding to this site, Zn2+ causes inhibition of [3H]1 methyl-4-phenylpyridinium ([3H]MPP+) uptake. We investigated the effect of Zn2+ on outward transport by superfusing hDAT-expressing HEK-293 cells preloaded with [3H]MPP+. Although Zn2+ inhibited uptake, Zn2+ facilitated [3H]MPP+ release induced by amphetamine, MPP+, or K+-induced depolarization specifically at hDAT but not at the human serotonin and the norepinephrine transporter (hNET). Mutation of the Zn2+ coordinating residue His(193) to Lys (the corresponding residue in hNET) eliminated the effect of Zn2+ on efflux. Conversely, the reciprocal mutation (K189H) conferred Zn2+ sensitivity to hNET. The intracellular [3H]MPP+ concentration was varied to generate saturation isotherms; these showed that Zn2+ increased V(max) for efflux (rather than K(M-Efflux-intracellular)). Thus, blockage of inward transport by Zn2+ is not due to a simple inhibition of the transporter turnover rate. The observations provide evidence against the model of facilitated exchange-diffusion and support the concept that inward and outward transport represent discrete operational modes of the transporter. In addition, they indicate a physiological role of Zn2+, because Zn2+ also facilitated transport reversal of DAT in rat striatal slices. PMID- 11940572 TI - ErbB-2 activates Stat3 alpha in a Src- and JAK2-dependent manner. AB - Stats (signal transducer and activator of transcription) are latent transcription factors that translocate from the cytoplasm to the nucleus. Constitutive activation of Stat3 alpha by upstream oncoproteins and receptor tyrosine kinases has been found in many human tumors and tumor-derived cell lines. Constitutively activated Stat3 alpha is often correlated with the activation of ErbB-2, a member of the EGFR family. To explore the involvement of ErbB-2 in the activation of Stat3 and the mechanism underlying this event, an ErbB-2 point mutant was used as a model of constitutively activated receptor. Phenylalanine mutations (Y-->F) were made in the autophosphorylation sites of the receptor, and their ability to activate Stat3 alpha was evaluated. Our results suggest that Stat3 alpha and JAK2 associates with ErbB-2 prior to phosphorylation of the receptor and that full activation of Stat3 alpha by ErbB-2 requires the participation of other non receptor tyrosine kinases. Both Src and Jak2 kinases contribute to the activation of Stat3 alpha but Src binds to ErbB-2 only when the receptor is phosphorylated. Our results also suggest that tyrosine 1139 may be important for Src homology 2 domain association because a mutant lacking this tyrosine reduces the ability of the Src homology 2 domain to bind to ErbB-2 and significantly decreases its ability to activate Stat3 alpha. PMID- 11940573 TI - Interactions between protein kinase CK2 and Pin1. Evidence for phosphorylation dependent interactions. AB - The peptidyl-prolyl isomerase Pin1 interacts in a phosphorylation-dependent manner with several proteins involved in cell cycle events. In this study, we demonstrate that Pin1 interacts with protein kinase CK2, an enzyme that generally exists in tetrameric complexes composed of two catalytic CK2 alpha and/or CK2 alpha' subunits together with two regulatory CK2 beta subunits. Our results indicate that Pin1 can interact with CK2 complexes that contain CK2 alpha. Furthermore, Pin1 can interact directly with the C-terminal domain of CK2 alpha that contains residues that are phosphorylated in vitro by p34(Cdc2) and in mitotic cells. Substitution of the phosphorylation sites of CK2 alpha with alanines resulted in decreased interactions between Pin1 and CK2. The other catalytic isoform of CK2, designated CK2 alpha', is not phosphorylated in mitotic cells and does not interact with Pin1, but a chimeric protein consisting of CK2 alpha' with the C terminus of CK2 alpha was phosphorylated in mitotic cells and interacts with Pin1, further implicating the phosphorylation sites in the interaction. In vitro, Pin1 inhibits the phosphorylation of Thr-1342 on human topoisomerase II alpha by CK2. Topoisomerase II alpha also interacts with Pin1 suggesting that the effect of Pin1 on the phosphorylation of Thr-1342 could result from its interactions with CK2 and/or topoisomerase II alpha. As compared with wild-type Pin1, isomerase-deficient and WW domain-deficient mutants of Pin1 are impaired in their ability to interact with CK2 and to inhibit the CK2 catalyzed phosphorylation of topoisomerase II alpha. Collectively, these results indicate that Pin1 and CK2 alpha interact and suggest a possible role for Pin1 in the regulation of topoisomerase II alpha. Furthermore, these results provide new insights into the functional role of the mitotic phosphorylation of CK2 and provide a new mechanism for selectively regulating the ability of CK2 to phosphorylate one of its mitotic targets. PMID- 11940574 TI - Activation of AXIN2 expression by beta-catenin-T cell factor. A feedback repressor pathway regulating Wnt signaling. AB - The Wnt pathway regulates cell fate, proliferation, and apoptosis, and defects in the pathway play a key role in many cancers. Although Wnts act to stabilize beta catenin levels in the cytosol and nucleus, a multiprotein complex containing adenomatous polyposis coli, glycogen synthase kinase 3beta, and Axin1 or its homolog Axin2/Axil/conductin promotes beta-catenin phosphorylation and subsequent proteasomal degradation. We found that the rat Axil gene was strongly induced upon neoplastic transformation of RK3E cells by mutant beta-catenin or gamma catenin or after ligand-induced activation of a beta-catenin-estrogen receptor fusion protein. Expression of Wnt1 in murine breast epithelial cells activated the conductin gene, and human cancers with defective beta-catenin regulation had elevated AXIN2 gene and protein expression. Expression of AXIN2/Axil was strongly repressed in cancer cells by restoration of wild type adenomatous polyposis coli function or expression of a dominant negative form of T cell factor (TCF)-4. TCF binding sites in the AXIN2 promoter played a key role in the ability of beta catenin to activate AXIN2 transcription. In contrast to AXIN2/Axil, expression of human or rat Axin1 homologs was nominally affected by beta-catenin-TCF. Because Axin2 can inhibit beta-catenin abundance and function, the data implicate AXIN2 in a negative feedback pathway regulating Wnt signaling. Additionally, although Axin1 and Axin2 have been thought to have comparable functions, the observation that Wnt pathway activation elevates AXIN2 but not AXIN1 expression suggests that there may be potentially significant functional differences between the two proteins. PMID- 11940575 TI - Direct involvement of CREB-binding protein/p300 in sequence-specific DNA binding of virus-activated interferon regulatory factor-3 holocomplex. AB - Infections of bacteria and viruses induce host defense reactions known as innate responses including the activation of interferon regulatory factor-3 (IRF-3), critical for the activation of type I interferon system. Upon immediate early signals triggered by the infection, IRF-3 is phosphorylated and a homodimer results. The homodimer complexes with the coactivator CREB-binding protein (CBP)/p300 in the nucleus; thus, holocomplex of IRF-3 competent in DNA binding is generated. We showed CBP/p300 to be indispensable for the DNA binding activity of the holocomplex and to aid the binding through direct interaction with the DNA. We demonstrated that p300 binds with the IRF-3 homodimer via a Q-rich domain and that an intact histone acetyltransferase (HAT) domain is indispensable for the DNA binding of the holocomplex along with a CH3 domain, which connects the HAT and Q-rich domains. These results highlight a novel function of CBP/p300: direct involvement in sequence-specific DNA binding. Furthermore, the critical function of these domains in virus-induced gene activation was demonstrated in vivo by using p300 mutants. PMID- 11940576 TI - Type I shorthorn sculpin antifreeze protein: recombinant synthesis, solution conformation, and ice growth inhibition studies. AB - A number of structurally diverse classes of "antifreeze" proteins that allow fish to survive in sub-zero ice-laden waters have been isolated from the blood plasma of cold water teleosts. However, despite receiving a great deal of attention, the one or more mechanisms through which these proteins act are not fully understood. In this report we have synthesized a type I antifreeze polypeptide (AFP) from the shorthorn sculpin Myoxocephalus scorpius using recombinant methods. Construction of a synthetic gene with optimized codon usage and expression as a glutathione S transferase fusion protein followed by purification yielded milligram amounts of polypeptide with two extra residues appended to the N terminus. Circular dichroism and NMR experiments, including residual dipolar coupling measurements on a 15N-labeled recombinant polypeptide, show that the polypeptides are alpha helical with the first four residues being more flexible than the remainder of the sequence. Both the recombinant and synthetic polypeptides modify ice growth, forming facetted crystals just below the freezing point, but display negligible thermal hysteresis. Acetylation of Lys-10, Lys-20, and Lys-21 as well as the N terminus of the recombinant polypeptide gave a derivative that displays both thermal hysteresis (0.4 degrees C at 15 mg/ml) and ice crystal faceting. These results confirm that the N terminus of wild-type polypeptide is functionally important and support our previously proposed mechanism for all type I proteins, in which the hydrophobic face is oriented toward the ice at the ice/water interface. PMID- 11940577 TI - Tumor necrosis factor-alpha increases airway smooth muscle oxidants production through a NADPH oxidase-like system to enhance myosin light chain phosphorylation and contractility. AB - Tumor necrosis factor plays a critical role in airway smooth muscle hyperresponsiveness observed in asthma. However, the mechanisms underlying this phenomenon are poorly understood. We investigated if tumor necrosis factor stimulated airway smooth muscle produced reactive oxygen species, leading to muscular hyperresponsiveness. Tumor necrosis factor increased intracellular and extracellular oxidants production in guinea pig airway smooth muscle cells and tissue homogenates. This production was abolished by inhibitors of NADPH oxidase (diphenylene iodinium or apocynin) and was enhanced by NADPH, whereas inhibitors of mitochondrial respiratory chain, nitric-oxide synthase, cyclooxygenase, and xanthine oxidase had no effect. NADPH oxidase subunits p22(phox) and p47(phox) were detected in smooth muscle cells and tissue homogenates by Western blot, immunohistochemistry, and spectral analysis. Furthermore, oxidants production was significantly reduced by transient transfection of smooth muscle cells with p22(phox) antisense oligonucleotides. Intracellular antioxidants and diphenylene iodinium abolished tumor necrosis factor-induced muscular hyperresponsiveness and increased in phosphorylation of the myosin light chain. Finally, NADPH oxidase subunits p22(phox) and p47(phox) were also detected in human airway smooth muscle. Collectively, these results demonstrate that tumor necrosis factor stimulated airway smooth muscle produces oxidants through a NADPH oxidase-like system, which plays a pivotal role in muscle hyperresponsiveness and myosin light chain phosphorylation. PMID- 11940578 TI - c-Raf/MEK/ERK pathway controls protein kinase C-mediated p70S6K activation in adult cardiac muscle cells. AB - p70S6 kinase (S6K1) plays a pivotal role in hypertrophic cardiac growth via ribosomal biogenesis. In pressure-overloaded myocardium, we show S6K1 activation accompanied by activation of protein kinase C (PKC), c-Raf, and mitogen-activated protein kinases (MAPKs). To explore the importance of the c-Raf/MAPK kinase (MEK)/MAPK pathway, we stimulated adult feline cardiomyocytes with 12-O tetradecanoylphorbol-13-acetate (TPA), insulin, or forskolin to activate PKC, phosphatidylinositol-3-OH kinase, or protein kinase A (PKA), respectively. These treatments resulted in S6K1 activation with Thr-389 phosphorylation as well as mammalian target of rapamycin (mTOR) and S6 protein phosphorylation. Thr-421/Ser 424 phosphorylation of S6K1 was observed predominantly in TPA-treated cells. Dominant negative c-Raf expression or a MEK1/2 inhibitor (U0126) treatment showed a profound blocking effect only on the TPA-stimulated phosphorylation of S6K1 and mTOR. Whereas p38 MAPK inhibitors exhibited only partial effect, MAPK-phosphatase 3 expression significantly blocked the TPA-stimulated S6K1 and mTOR phosphorylation. Inhibition of mTOR with rapamycin blocked the Thr-389 but not the Thr-421/Ser-424 phosphorylation of S6K1. Therefore, during PKC activation, the c-Raf/MEK/extracellular signal-regulated kinase-1/2 (ERK1/2) pathway mediates both the Thr-421/Ser-424 and the Thr-389 phosphorylation in an mTOR-independent and -dependent manner, respectively. Together, our in vivo and in vitro studies indicate that the PKC/c-Raf/MEK/ERK pathway plays a major role in the S6K1 activation in hypertrophic cardiac growth. PMID- 11940579 TI - Insulin-like growth factor-binding protein-3 activates a phosphotyrosine phosphatase. Effects on the insulin-like growth factor signaling pathway. AB - The proliferative action of insulin-like growth factors (IGF-I and -II) is mediated via the type I IGF receptor (IGF-IR) and is modulated by their association with high affinity binding proteins, IGFBP-1 to -6. We recently found that, in addition to its ability to bind IGFs, IGFBP-3 also inhibits IGF-IR activation independently of IGF binding and without interacting directly with IGF IR. Here, we show that IGFBP-3 is capable of blocking the signal triggered by IGFs. Breast carcinoma-derived cells (MCF-7) were stimulated by des(1-3)IGF-I or [Gln(3),Ala(4),Tyr(15),Leu(16)]IGF-I, two IGF analogues with intact affinity for IGF-IR, but with weak or virtually no affinity for IGFBPs, then incubated with IGFBP-3. The activated IGF-IR was desensitized through reversal of its autophosphorylation, following which both phosphatidylinositol 3-kinase and p42(MAPK) activities were depressed. Direct measurement of phosphotyrosine phosphatase activity and reconstitution experiments using tyrosine-phosphorylated insulin receptor substrate-1 (IRS-1) indicated that IGFBP-3 activated a phosphotyrosine phosphatase (PTPase). This action appeared to be peculiar to IGFBP-3 among the IGFBPs, since neither IGFBP-1 nor IGFBP-5 (structurally the closest to IGFBP-3), had any such effect. Several cell lines derived from normal or tumor cells responsive to IGF-I were used to show that IGFBP-3-stimulated PTPase is cell type-specific. Although the precise nature of the phosphatase remains to be determined, the results of this study demonstrate that IGFBP-3 stimulates a phosphotyrosine phosphatase activity that down-regulates the IGF-I signaling pathway, suggesting a major role for IGFBP-3 in regulating cell proliferation. PMID- 11940580 TI - HIV-1 gp41 envelope residues 650-685 exposed on native virus act as a lectin to bind epithelial cell galactosyl ceramide. AB - The initial step in the interaction between human immunodeficiency virus (HIV-1) and epithelial cells is the binding of HIV-1 envelope glycoproteins to the epithelial cell galactosyl ceramide (GalCer). Here we show that HIV-1 envelope gp41 residues 650-685 bind GalCer in a galactose-specific manner. The gp41 residues that display this lectin activity are highly conserved among HIV-1 isolates and constitute three regions: residues 650-661, which encompass a charged helix; residues 662-667, referred to as the conserved epitope ELDKWA, the epitope recognized by antibodies that neutralize HIV-1 entry in epithelial and CD4(+)-mononucleated cells; and residues 668-685, a hydrophobic Trp-rich sequence that stabilizes the structure of the galactose binding site. Similar to other galactose-specific lectins, the gp41 lectin site is active only as an oligomer. Finally the orientation of the galactose toward the gp41 lectin site appears to be controlled by the lipid microenvironment of the epithelial membrane. From the experimental data we construct a theoretical model of the interaction between gp41 and GalCer based on thermodynamic considerations. This model integrates the dynamics and the spatial organization of the viral envelope glycoproteins, GalCer organized in raft microdomains in the apical region of the epithelial cell membrane and the interfacial water. Characterization of the minimal sequence and structure of gp41 in direct interaction with GalCer may help unravel the still unknown immunogenic determinant able to elicit antibodies against ELDKWA and target of one of the rare neutralizing antibodies against gp41. PMID- 11940581 TI - Protein kinase C-alpha and protein kinase C-epsilon are required for Grb2 associated binder-1 tyrosine phosphorylation in response to platelet-derived growth factor. AB - Grb2-associated binder-1 (Gab1) is an adapter protein related to the insulin receptor substrate family. It is a substrate for the insulin receptor as well as the epidermal growth factor (EGF) receptor and other receptor-tyrosine kinases. To investigate the role of Gab1 in signaling pathways downstream of growth factor receptors, we stimulated rat aortic vascular smooth muscle cells (VSMC) with EGF and platelet-derived growth factor (PDGF). Gab1 was tyrosine-phosphorylated by EGF and PDGF within 1 min. AG1478 (an EGF receptor kinase-specific inhibitor) failed to block PDGF-induced Gab1 tyrosine phosphorylation, suggesting that transactivated EGF receptor is not responsible for this signaling event. Because Gab1 associates with phospholipase Cgamma (PLCgamma), we studied the role of the PLCgamma pathway in Gab1 tyrosine phosphorylation. Gab1 tyrosine phosphorylation by PDGF was impaired in Chinese hamster ovary cells expressing mutant PDGFbeta receptor (Y977F/Y989F: lacking the binding site for PLCgamma). Pretreatment of VSMC with (a specific PLCgamma inhibitor) inhibited Gab1 tyrosine phosphorylation as well, indicating the importance of the PLCgamma pathway. Gab1 was tyrosine phosphorylated by phorbol ester to the same extent as PDGF stimulation. Studies using antisense protein kinase C (PKC) oligonucleotides and specific inhibitors showed that PKCalpha and PKCepsilon are required for Gab1 tyrosine phosphorylation. Binding of Gab1 to the protein-tyrosine phosphatase SHP2 and phosphatidylinositol 3-kinase was significantly decreased by PLCgamma and/or PKC inhibition, suggesting the importance of the PLCgamma/PKC-dependent Gab1 tyrosine phosphorylation for the interaction with other signaling molecules. Because PDGF mediated ERK activation is enhanced in Chinese hamster ovary cells that overexpress Gab1, Gab1 serves as an important link between PKC and ERK activation by PDGFbeta receptors in VSMC. PMID- 11940582 TI - The kinetic mechanism of Myo1e (human myosin-IC). AB - Myo1e is the widely expressed subclass-1 member of the myosin-I family. We performed a kinetic analysis of a truncated myo1e that consists of the motor and the single IQ motif with a bound calmodulin. We determined the rates and equilibrium constants for the key steps in the ATPase cycle. The maximum actin activated ATPase rate (V(max)) and the actin concentration at half-maximum of V(max) (K(ATPase)) of myo1e are similar to those of the native protein. The K(ATPase) is low (approximately 1 microm), however the affinity of myo1e for actin in the presence of ATP is very weak. A weak actin affinity and a rapid rate of phosphate release result in a pathway under in vitro assay conditions in which phosphate is released while myo1e is dissociated from actin. Actin activation of the ATPase activity and the low K(ATPase) are the result of actin activation of ADP release. We propose that myo1e is tuned to function in regions of high concentrations of cross-linked actin filaments. Additionally, we found that ADP release from actomyo1e is > 10-fold faster than other vertebrate myosin-I isoforms. We propose that subclass-1 myosin-Is are tuned for rapid sliding, whereas subclass-2 isoforms are tuned for tension maintenance or stress sensing. PMID- 11940583 TI - Monitoring of ligand-independent dimerization and ligand-induced conformational changes of melatonin receptors in living cells by bioluminescence resonance energy transfer. AB - Several G protein-coupled receptors have been shown to exist as homo-and hetero oligomeric complexes in living cells. However, the link between ligand-induced receptor activation and its oligomerization state as well as the proportion of the total receptor population that can engage in oligomeric complexes remain open questions. Here, the closely related human MT1 and MT2 melatonin receptors (MT1R, MT2R) were used to address these issues. Bioluminescence resonance energy transfer (BRET) experiments in living HEK 293 cells revealed that these receptors form homo- and hetero-oligomers. Constitutive energy transfer was observed for all receptor combinations at physiological expression levels and could be detected in single cell BRET experiments. Inhibition of the energy transfer by dilution of the BRET partners identified MT1R and MT2R dimers as the predominant receptor species, and this oligomerization state did not change upon agonist and antagonist binding. Agonists, neutral antagonists, and inverse agonists all promoted increases in BRET values for MT2R but not for MT1R homodimers in living cells and isolated plasma membranes. This indicates that no correlation could be inferred between the receptor activation state and the dimerization state of the receptor. This also suggests that ligand-promoted BRET increases represent specific ligand-induced conformational changes of pre-existing dimers rather then increased dimerization. The observation that ligands favored the energy transfer within the hetero-oligomer from MT1R to MT2R but not in the reverse orientation, from MT2R to MT1R, supports this view. PMID- 11940584 TI - Crystal structure of a dual activity IMPase/FBPase (AF2372) from Archaeoglobus fulgidus. The story of a mobile loop. AB - Several hyperthermophilic organisms contain an unusual phosphatase that has dual activity toward inositol monophosphates and fructose 1,6-bisphosphate. The structure of the second member of this family, an FBPase/IMPase from Archaeoglobus fulgidus (AF2372), has been solved. This enzyme shares many kinetic and structural similarities with that of a previously solved enzyme from Methanococcus jannaschii (MJ0109). It also shows some kinetic differences in divalent metal ion binding as well as structural variations at the dimer interface that correlate with decreased thermal stability. The availability of different crystal forms allowed us to investigate the effect of the presence of ligands on the conformation of a mobile catalytic loop independently of the crystal packing. This conformational variability in AF2372 is compared with that observed in other members of this structural family that are sensitive or insensitive to submillimolar concentrations of Li(+). This analysis provides support for the previously proposed mechanism of catalysis involving three metal ions. A direct correlation of the loop conformation with strength of Li(+) inhibition provides a useful system of classification for this extended family of enzymes. PMID- 11940585 TI - Solution structure of the orphan PABC domain from Saccharomyces cerevisiae poly(A)-binding protein. AB - We have determined the solution structure of the PABC domain from Saccharomyces cerevisiae Pab1p and mapped its peptide-binding site. PABC domains are peptide binding domains found in poly(A)-binding proteins (PABP) and are a subset of HECT family E3 ubiquitin ligases (also known as hyperplastic discs proteins (HYDs)). In mammals, the PABC domain of PABP functions to recruit several different translation factors to the mRNA poly(A) tail. PABC domains are highly conserved, with high specificity for peptide sequences of roughly 12 residues with conserved alanine, phenylalanine, and proline residues at positions 7, 10, and 12. Compared with human PABP, the yeast PABC domain is missing the first alpha helix, contains two extra amino acids between helices 2 and 3, and has a strongly bent C-terminal helix. These give rise to unique peptide binding specificity wherein yeast PABC binds peptides from Paip2 and RF3 but not Paip1. Mapping of the peptide-binding site reveals that the bend in the C-terminal helix disrupts binding interactions with the N terminus of peptide ligands and leads to greatly reduced binding affinity for the peptides tested. No high affinity or natural binding partners from S. cerevisiae could be identified by sequence analysis of known PABC ligands. Comparison of the three known PABC structures shows that the features responsible for peptide binding are highly conserved and responsible for the distinct but overlapping binding specificities. PMID- 11940586 TI - Ligand specificity and conformational dependence of the hepatic nuclear factor 4alpha (HNF-4alpha ). AB - Hepatic nuclear factor-4alpha (HNF-4alpha) controls the expression of genes encoding proteins involved in lipid and carbohydrate metabolism. Fatty acyl-CoA thioesters have recently been proposed to be naturally occurring ligands of HNF 4alpha and to regulate its transcriptional activity as function of their chain length and degree of unsaturation (Hertz, R., Magenheim, J., Berman, I., and Bar Tana, J. (1998) Nature 392, 512-516). However, the apparent low affinities (microm K(d) values) obtained with a radiolabeled fatty acyl-CoA ligand binding assay raised questions regarding the physiological significance of this finding. Furthermore, it is not known whether interaction with fatty acyl-CoA alters the structure of HNF-4alpha. These issues were examined using rat recombinant HNF 4alpha ligand-binding domain (HNF-4alphaLBD) in conjunction with photon counting fluorescence and circular dichroism. First, fluorescence resonance energy transfer between HNF-4alphaLBD tryptophan (Trp) and cis-parinaroyl-CoA yielded an intermolecular distance of 0.95) and neck length (r (2) > 0.75). Increasing body weight affected the peak cement strains far more than did increasing neck length. During simulated stair climbing, calcar-collar contact reduced peak strains in the proximal cement by a factor of 1.5 to two. Peak principal tensile strains in the proximal cement often exceeded 1000 me when the smaller stems were used. CONCLUSIONS: In this stair climbing test model, the peak proximal cement strains were increased more by changes in body weight than they were by changes in neck length. Even during stair-climbing, calcar-collar contact reduced peak cement strains. PMID- 11940618 TI - Anterior knee pain after intramedullary nailing of fractures of the tibial shaft. A prospective, randomized study comparing two different nail-insertion techniques. AB - BACKGROUND: Anterior knee pain is the most common complication after intramedullary nailing of the tibia. Dissection of the patellar tendon and its sheath during nailing is thought to be a contributing cause of chronic anterior knee pain. The purpose of this prospective, randomized study was to assess whether the prevalence or the intensity of anterior knee pain following intramedullary nailing of a tibial shaft fracture is reduced by the use of a paratendinous incision for the nail entry portal. METHODS: Fifty patients with a tibial shaft fracture requiring intramedullary nailing were randomized equally to treatment with paratendinous or transtendinous nailing. Twenty-one patients from both study groups were followed for an average of three years after nailing. After fracture union, all but two patients had elective nail removal through the same surgical approach as was used for the nailing. At the follow-up evaluation, the patients used visual analog scales to report their level of anterior knee pain and the impairment caused by that pain. The scales described by Lysholm and Gillquist and by Tegner et al., the Iowa knee scoring system, and simple functional tests were used to quantitate the functional results. Isokinetic thigh muscle strength was also measured. RESULTS: Fourteen (67%) of the twenty-one patients treated with transtendinous nailing reported anterior knee pain at the final evaluation. Of these fourteen patients, thirteen were mildly to severely impaired by the pain. Fifteen (71%) of the twenty-one patients treated with paratendinous nailing reported anterior knee pain, and ten of the fifteen were impaired by the pain. The Lysholm, Tegner, and Iowa knee scoring systems; muscle strength measurements; and functional tests showed no significant differences between the two groups. CONCLUSION: Compared with a transpatellar tendon approach, a paratendinous approach for nail insertion does not reduce the prevalence of chronic anterior knee pain or functional impairment by a clinically relevant amount after intramedullary nailing of a tibial shaft fracture. PMID- 11940619 TI - Comparison of clinical outcomes in total hip arthroplasty using rough and polished cemented stems with essentially the same geometry. AB - BACKGROUND: Aseptic loosening of the cemented stem is the most common cause of revision of total hip arthroplasties. The loosening is often associated with substantial lysis of the surrounding bone. The surface finish of femoral components is suspected as a contributing factor to this bone lysis. The purpose of this study was to compare the results associated with a rough surface and those associated with a polished surface in a consecutive series of cemented stems with essentially the same geometry. METHODS: The study included 244 consecutive total hip arthroplasties with a cemented femoral component performed by one surgeon. There was no difference in patient selection criteria or surgical techniques between the group treated with a polished stem and that treated with a grit-blasted stem. All arthroplasties were hybrid, with an uncemented acetabular component. Generally, patients were over the age of sixty years (mean age, 70.6 years). The stems inserted in the initial 122 hips had a grit-blasted surface with a roughness of 2.1 microm. The stem surface in the second 122 hips was polished (roughness, 0.1 microm). The results of clinical and radiographic assessments performed immediately after surgery were compared with those performed at the most recent visit. The average duration of clinical follow-up for the patients treated with the grit-blasted and polished stems was 5.98 years and 5.32 years, respectively. RESULTS: Four hips treated with the grit-blasted stem had aseptic loosening with substantial surrounding lysis and required revision. An additional two hips in this group had radiographic evidence of substantial lysis and were judged to have an impending need for revision. In contrast, no hip treated with the polished stem required revision, and only one had minimal lysis. This difference regarding failures and impending failures was significant (p = 0.05). The clinical results were comparable, with an Iowa hip rating of 98 points at the time of follow-up in both groups. CONCLUSIONS: There was a significant difference between grit-blasted and polished stems with respect to the prevalence of revisions and impending revisions, all of which were identified in a relatively short follow-up period. The results in this series favor the use of a polished stem when cement is employed for fixation of the femoral component. PMID- 11940620 TI - Long-standing nonunion of fractures of the lateral humeral condyle. AB - BACKGROUND: Patients with nonunion of a fracture of the lateral humeral condyle often have pain, instability, or progressive cubitus valgus deformity with tardy ulnar nerve palsy. However, some patients have minimal or no symptoms or disabilities. We evaluated patients with long-standing established nonunion of the lateral humeral condyle to correlate the clinical long-term outcome of this condition with the original fracture type. METHODS: Nineteen elbows in eighteen patients who were at least twenty years of age were evaluated. Fourteen patients were male, and four were female. The average age at presentation was 42.5 years. The average interval from the injury to the presentation of the symptoms of the nonunion was thirty-seven years. Patients were divided into two groups on the basis of the size of the fragment and the location of the fracture line. Group 1 included nine elbows with nonunion resulting from a Milch Type-I injury, and Group 2 included ten elbows with a nonunion resulting from a Milch Type-II injury. Evaluations were performed with use of radiographic examination, clinical assessment, and calculation of the Broberg and Morrey score. RESULTS: Symptoms were seen more frequently in Group 1 than in Group 2. The range of flexion in Group 1 (range, 60 degrees to 145 degrees; average, 99 degrees) was more restricted than that in Group 2 (range, 100 degrees to 150 degrees; average, 129 degrees) (p = 0.0078). The functional score in Group 2 was significantly higher than that in Group 1 (p = 0.03). CONCLUSION: Disabling symptoms only rarely developed in Group-2 patients. Occasionally, however, these patients do present with clinically detectable dysfunction of the ulnar nerve. In contrast, pain, instability, and loss of range of motion as well as ulnar nerve dysfunction developed in Group 1. For this reason we think that a nonunion of a Milch Type-I fracture should be treated as soon as possible after injury, preferably before the patient reaches skeletal maturity. PMID- 11940621 TI - Total knee arthroplasty for osteonecrosis. AB - BACKGROUND: A patient with collapse of a femoral condyle caused by osteonecrosis has few treatment options other than total knee arthroplasty. The purpose of this study was to report the clinical and radiographic outcome of total knee arthroplasty for osteonecrosis. METHODS: Between 1987 and 1996, thirty-two total knee arthroplasties were performed with cement in thirty patients with osteonecrosis of the femoral condyle and/or tibial plateau. The study group included twenty-forty women and five men with a mean age of fifty-four years (range, thirty-one to seventy-seven years) at the time of the arthroplasty. Twenty-two patients had atraumatic osteonecrosis associated with corticosteroid use, and eight had spontaneous osteonecrosis. All patients had a complete clinical and radiographic evaluation at a mean of 108 months (range, forty-eight to 144 months) postoperatively. RESULTS: Overall, thirty-one (97%) of the thirty two knees had a successful clinical outcome. The mean Knee Society score improved from 54 points preoperatively to 95 points at the time of the latest follow-up. No evidence of progressive radiolucency was found around any prosthetic component. CONCLUSIONS: Previous studies have demonstrated less-than-optimal results following total knee arthroplasty in patients with osteonecrosis. The excellent results found in the present study may have been secondary to the use of cemented implants in all cases and ancillary stems when appropriate. PMID- 11940622 TI - Educational deficiencies in musculoskeletal medicine. AB - BACKGROUND: We previously reported the results of a study in which a basic competency examination in musculoskeletal medicine was administered to a group of recent medical school graduates. This examination was validated by 124 orthopaedic program directors, and a passing grade of 73.1% was established. According to that criterion, 82% of the examinees failed to demonstrate basic competency in musculoskeletal medicine. It was suggested that perhaps a different passing grade would have been set by program directors of internal medicine departments. To test that hypothesis, and to determine whether the importance of the individual questions would be rated similarly, the validation process was repeated with program directors of internal medicine residency departments as subjects. METHODS: Our basic competency examination was sent to all 417 program directors of internal medicine departments in the United States. Each recipient was mailed a letter of introduction explaining the purpose of the study, a copy of the examination, and our answer key and scoring guide. There was no mention of the results of the first study. The subjects were requested to rate the importance of each question on the same visual analog scale, ranging from "not important" to "very important," as had been used by the orthopaedic program directors. These ratings were converted into numerical scores. The program directors were also asked to suggest a passing score for the examination, and this score was used to assess the examinees' performance on the examination. The results on the basis of the internal medicine program directors' responses and those according to the orthopaedic program directors' responses were compared. RESULTS: Two hundred and forty (58%) of the 417 program directors of internal medicine residency departments responded. They suggested a mean passing score (and standard deviation) of 70.0% +/- 9.9%. As reported previously, the mean test score of the eighty-five examinees was 59.6%. Sixty-six (78%) of them failed to demonstrate basic competency on the examination according to the criterion set by the internal medicine program directors. The internal medicine program directors assigned a mean importance score of 7.4 (of 10) to the questions on the examination compared with a mean score of 7.0 assigned by the orthopaedic program directors. The internal medicine program directors gave twenty-four of the twenty five questions an importance score of at least 5 and seventeen of the twenty-five questions an importance score of at least 6.6. CONCLUSIONS: According to the standard suggested by the program directors of internal medicine residency departments, a large majority of the examinees once again failed to demonstrate basic competency in musculoskeletal medicine on the examination. It is therefore reasonable to conclude that medical school preparation in musculoskeletal medicine is inadequate. PMID- 11940623 TI - Use of helical computed tomography for the assessment of acetabular osteolysis after total hip arthroplasty. AB - BACKGROUND: Acetabular osteolysis is a major problem affecting long-term survival of total hip prostheses. Since lytic lesions may be asymptomatic until extensive bone loss has occurred, early detection of lytic lesions is important. The purposes of this study were to determine the efficacy and potential role of high resolution helical (or spiral) computed tomography with metal-artifact minimization in the early detection of osteolysis of the pelvis and to use the method to determine if there was a relationship between the extent of osteolysis and the amount of polyethylene wear. METHODS: Forty patients (fifty hips) who had undergone primary cementless total hip arthroplasty between 1988 and 1994 were evaluated as part of an ongoing prospective study. These patients had a history of high-level activity that was believed to place them at increased risk for accelerated polyethylene wear. The most recent follow-up radiographs were compared with the three-month postoperative radiographs. Helical computed tomography scans with metal-artifact minimization were made, and evidence of osteolytic lesions on these scans was compared with that on the radiographs. Two dimensional wear analysis was performed with use of digitized radiographs, and the results were compared with loss of bone volume as calculated from the computed tomography scans. RESULTS: Acetabular lysis was identified on the radiographs of sixteen hips and on the computed tomography scans of twenty-six hips. Radiographs underestimated the extent of the lysis in thirteen of the sixteen hips. There was no correlation (r = 0.036) between linear wear and the measured volume of bone loss, with the numbers available. On the basis of the amount of lysis seen on the computed tomography scans, one patient underwent a revision procedure. CONCLUSIONS: Helical computed tomography with metal-artifact minimization is more sensitive for identifying and quantifying osteolysis after total hip arthroplasty than is plain radiography. Since computed tomography scans show both the extent and the location of lytic lesions, they are useful to guide treatment decisions as well as to assist in planning for surgical intervention, when needed, in patients with suspected osteolysis. PMID- 11940624 TI - An observational study of orthopaedic abstracts and subsequent full-text publications. AB - BACKGROUND: Research abstracts are frequently referenced in orthopaedic textbooks and influence orthopaedic care. However, little is known about the quality of information provided in the abstracts, the frequency of publication of complete papers after presentation of abstracts, or any discrepancies between abstracts and published papers. The objective of this study was to determine the quality of information provided in orthopaedic abstracts, rates of publication of full-text articles after presentation of abstracts, predictors of publication of full-text articles, and consistency between abstracts and full-text articles. METHODS: We retrieved all abstracts from the 1996 scientific program of the sixty-third Annual Meeting of the American Academy of Orthopaedic Surgeons. For each abstract, we recorded the completeness of reporting and key features of the study design, conduct, analysis, and interpretation. A computerized Medline and PubMed search established whether the abstract had been followed by publication of a full-text article. Finally, we evaluated the consistency of reporting between abstracts and final publications. RESULTS: The program included 465 abstracts, 66% of which were on prognostic studies. All abstracts described the study design, and 70.7% of the designs were observational. Key methodological issues were reported in less than half of the abstracts, and information on data analysis was reported in <15%. One hundred and fifty-nine (34%) of the 465 abstracts were followed by publication of a full-text article. The mean time to publication (and standard deviation) was 17.6 +/- 12 months (range, one to fifty six months). Inconsistencies between the abstract and the full-text article included the primary outcome measure, which differed 14% of the time, and the results, which differed 19% of the time. CONCLUSIONS: Two-thirds of the orthopaedic abstracts in this sample were not followed by publication of a full text paper. The overall quality of reporting in abstracts proved inadequate, and inconsistencies between the final published paper and the original abstract occurred frequently. The routine use of abstracts as a guide to orthopaedic practice needs to be reconsidered. PMID- 11940625 TI - Subluxation of the sternoclavicular joint secondary to pseudarthrosis of the first and second ribs. A case report. PMID- 11940626 TI - Zoonotic onchocerciasis of the shoulder. A case report. PMID- 11940627 TI - Membranous lipodystrophy. A case report. PMID- 11940628 TI - Jaffe-Campanacci syndrome. A case report and review of the literature. PMID- 11940629 TI - Catastrophic fat embolism following augmentation of pedicle screws with bone cement : a case report. PMID- 11940632 TI - ICOE welcomes JBJS and OREF as partners. PMID- 11940633 TI - National Consensus Conference on Improving the Continuum of Care for Patients with Hip Fracture. PMID- 11940634 TI - Electromagnetic surgical devices in orthopaedics. Lasers and radiofrequency. PMID- 11940635 TI - JSBS classics. Prevention of infection in the treatment of one thousand and twenty-five open fractures of long bones. Retrospective and prospective analyses. PMID- 11940636 TI - Measuring cognitive and clinical competency in orthopaedics. PMID- 11940637 TI - Osteoporosis awareness: implications for improving the care of patients with hip fracture. PMID- 11940638 TI - Should the term "required" be banned? PMID- 11940639 TI - Cervical spine growth in children. PMID- 11940640 TI - Stress radiographs are important in diagnosing valgus instability of the elbow. PMID- 11940641 TI - Questions about intramedullary nailing. PMID- 11940643 TI - Discerning the merit of "significant results". PMID- 11940644 TI - Cement leakage and the need for prophylactic fenestration of the spinal canal during vertebroplasty. PMID- 11940645 TI - Assessing a patient's capacity to refuse treatment. PMID- 11940646 TI - What's new in musculoskeletal oncology. PMID- 11940647 TI - Enhanced expression of the transcription factor Nrf2 by cancer chemopreventive agents: role of antioxidant response element-like sequences in the nrf2 promoter. AB - Induction of phase 2 enzymes, which neutralize reactive electrophiles and act as indirect antioxidants, is an important mechanism for protection against carcinogenesis. The transcription factor Nrf2, which binds to the antioxidant response element (ARE) found in the upstream regulatory region of many phase 2 genes, is essential for the induction of these enzymes. We have investigated the effect of the potent enzyme inducer and anticarcinogen 3H-1,2-dithiole-3-thione (D3T) on the fate of Nrf2 in murine keratinocytes. Both total and nuclear Nrf2 levels increased rapidly and persistently after treatment with D3T but could be blocked by cotreatment with cycloheximide. Nrf2 mRNA levels increased approximately 2-fold 6 h after D3T treatment. To examine the transcriptional activation of Nrf2 by D3T, the proximal region (1 kb) of the nrf2 promoter was isolated. Deletion and mutagenesis analyses demonstrated that nrf2 promoter luciferase reporter activity was enhanced by treatment with D3T and that ARE-like sequences were required for this activation. Gel shift assays with nuclear extracts from PE cells indicated that common factors bind to typical AREs and the ARE-like sequences of the nrf2 promoter. Direct binding of Nrf2 to its own promoter was demonstrated by chromatin immunoprecipitation assay. Overexpression of Nrf2 increased the activity of the nrf2 promoter-luciferase reporter, while expression of mutant Nrf2 protein repressed activity. Thus, Nrf2 appears to autoregulate its own expression through an ARE-like element located in the proximal region of its promoter, leading to persistent nuclear accumulation of Nrf2 and protracted induction of phase 2 genes in response to chemopreventive agents. PMID- 11940648 TI - The MyoD-inducible p204 protein overcomes the inhibition of myoblast differentiation by Id proteins. AB - The murine p204 protein level is highest in heart and skeletal muscle. During the fusion of cultured myoblasts to myotubes, the p204 level increases due to transcription dependent on the muscle-specific MyoD protein, and p204 is phosphorylated and translocated from the nucleus to the cytoplasm. p204 overexpression accelerates myoblast fusion in differentiation medium and triggers this process even in growth medium. Here we report that p204 is required for the differentiation of C2C12 myoblasts. We propose that it enables the differentiation, at least in part, by overcoming the inhibition of the activities of the MyoD and E47 proteins by the Id proteins: Id1, Id2, and Id3. These are known to inhibit skeletal muscle differentiation by binding and blocking the activity of MyoD, E12/E47, and other myogenic basic helix-loop-helix (bHLH) proteins. Our hypothesis is based on the following findings. (i) A decrease in the p204 level in C2C12 myoblasts by antisense RNA (a) increased the level of the Id2; (b) inhibited the MyoD-, E12/E47-, and other bHLH protein-dependent accumulation of the muscle-specific myosin heavy-chain protein; and (c) inhibited the fusion of myoblasts to myotubes in differentiation medium. (ii) p204 bound to the Id proteins in vitro and in vivo. (iii) In the binding of p204 to Id2, the b segment of p204 and the HLH segment of Id2 were involved. (iv) Addition of p204 overcame the inhibition by the Id proteins of the binding of MyoD and E47 to DNA in vitro. (v) Overexpression of p204 in myoblasts (a) decreased the level of the Id proteins, even in a culture in growth medium, and (b) overcame the inhibition by the Id proteins of MyoD- and E47 dependent transcription and also overcame the inhibition by Id2 of the fusion of myoblasts to myotubes. PMID- 11940650 TI - Evidence that negative elongation factor represses transcription elongation through binding to a DRB sensitivity-inducing factor/RNA polymerase II complex and RNA. AB - Negative elongation factor (NELF) is a human transcription factor complex that cooperates with DRB sensitivity-inducing factor (DSIF)/hSpt4-hSpt5 to repress elongation by RNA polymerase II (RNAPII). NELF activity is associated with five polypeptides, including NELF-A, a candidate gene product for Wolf-Hirschhorn syndrome, and NELF-E, a putative RNA-binding protein with arginine-aspartic acid (RD) dipeptide repeats. Here we report several important findings regarding the DSIF/NELF-dependent elongation control. First, we have established an effective method for purifying the active NELF complex using an epitope-tagging technique. Second, the five polypeptides each are important and together are sufficient for its function in vitro. Third, NELF does not bind to either DSIF or RNAPII alone but does bind to the preformed DSIF/RNAPII complex. Fourth, NELF-E has a functional RNA-binding domain, whose mutations impair transcription repression without affecting known protein-protein interactions. Taken together, we propose that NELF causes RNAPII pausing through binding to the DSIF/RNAPII complex and to nascent transcripts. These results also have implications for how DSIF and NELF are regulated in a gene-specific manner in vivo. PMID- 11940649 TI - DNA methyltransferase deficiency modifies cancer susceptibility in mice lacking DNA mismatch repair. AB - We have introduced DNA methyltransferase 1 (Dnmt1) mutations into a mouse strain deficient for the Mlh1 protein to study the interaction between DNA mismatch repair deficiency and DNA methylation. Mice harboring hypomorphic Dnmt1 mutations showed diminished RNA expression and DNA hypomethylation but developed normally and were tumor free. When crossed to Mlh1(-/-) homozygosity, they were less likely to develop the intestinal cancers that normally arise in this tumor predisposed, mismatch repair-deficient background. However, these same mice developed invasive T- and B-cell lymphomas earlier and at a much higher frequency than their Dnmt1 wild-type littermates. Thus, the reduction of Dnmt1 activity has significant but opposing outcomes in the development of two different tumor types. DNA hypomethylation and mismatch repair deficiency interact to exacerbate lymphomagenesis, while hypomethylation protects against intestinal tumors. The increased lymphomagenesis in Dnmt1 hypomorphic, Mlh1(-/-) mice may be due to a combination of several mechanisms, including elevated mutation rates, increased expression of proviral sequences or proto-oncogenes, and/or enhanced genomic instability. We show that CpG island hypermethylation occurs in the normal intestinal mucosa, is increased in intestinal tumors in Mlh1(-/-) mice, and is reduced in the normal mucosa and tumors of Dnmt1 mutant mice, consistent with a role for Dnmt1-mediated CpG island hypermethylation in intestinal tumorigenesis. PMID- 11940651 TI - P53 is necessary for the apoptotic response mediated by a transient increase of Ras activity. AB - The tumor suppressor p53 eliminates cancer-prone cells via multiple mechanisms, including apoptosis. Ras elicits apoptosis in cells after protein kinase C (PKC) downregulation. However, the role of p53 in Ras-mediated apoptosis has not been fully investigated. Here, we demonstrate that mouse fibroblasts that express wild type p53 are more susceptible to apoptosis elicited by PKC inhibition if Ras is transiently expressed or upregulated as opposed to stably expressed. In the latter case, p53 is frequently mutated. Transiently increased Ras activity induces Bax, and PKC inhibition augments this induction. Overexpression of E6 inactivates p53 and thereby suppresses both Bax induction and apoptosis. In contrast, Bax is not induced in stable ras transfectants, regardless of PKC inhibition. The data suggest that short- and long-term activation of Ras use a different mechanism(s) to initiate apoptosis. The status of p53 may contribute to such differences. PMID- 11940652 TI - Cdc42 regulation of kinase activity and signaling by the yeast p21-activated kinase Ste20. AB - The Saccharomyces cerevisiae kinase Ste20 is a member of the p21-activated kinase (PAK) family with several functions, including pheromone-responsive signal transduction. While PAKs are usually activated by small G proteins and Ste20 binds Cdc42, the role of Cdc42-Ste20 binding has been controversial, largely because Ste20 lacking its entire Cdc42-binding (CRIB) domain retains kinase activity and pheromone response. Here we show that, unlike CRIB deletion, point mutations in the Ste20 CRIB domain that disrupt Cdc42 binding also disrupt pheromone signaling. We also found that Ste20 kinase activity is stimulated by GTP-bound Cdc42 in vivo and this effect is blocked by the CRIB point mutations. Moreover, the Ste20 CRIB and kinase domains bind each other, and mutations that disrupt this interaction cause hyperactive kinase activity and bypass the requirement for Cdc42 binding. These observations demonstrate that the Ste20 CRIB domain is autoinhibitory and that this negative effect is antagonized by Cdc42 to promote Ste20 kinase activity and signaling. Parallel results were observed for filamentation pathway signaling, suggesting that the requirement for Cdc42-Ste20 interaction is not qualitatively different between the mating and filamentation pathways. While necessary for pheromone signaling, the role of the Cdc42-Ste20 interaction does not require regulation by pheromone or the pheromone-activated G beta gamma complex, because the CRIB point mutations also disrupt signaling by activated forms of the kinase cascade scaffold protein Ste5. In total, our observations indicate that Cdc42 converts Ste20 to an active form, while pathway stimuli regulate the ability of this active Ste20 to trigger signaling through a particular pathway. PMID- 11940653 TI - Socius is a novel Rnd GTPase-interacting protein involved in disassembly of actin stress fibers. AB - Rho family small GTPases are key regulators of the actin cytoskeleton in various cell types. The Rnd proteins, Rnd1, Rnd2, and Rnd3/RhoE, have been recently identified as new members of the Rho family of GTPases, and expression of Rnd1 or Rnd3 in fibroblasts causes the disassembly of actin stress fibers and the retraction of the cell body to produce extensively branching cellular processes. Here we have performed a yeast two-hybrid screening by using Rnd1 as bait and identified a novel protein that specifically binds to Rnd GTPases. We named this protein Socius. Socius directly binds to Rnd GTPases through its COOH-terminal region. When transfected into COS-7 cells, Socius is translocated to the cell periphery in response to Rnd1 and Rnd3 and colocalized with the GTPases. While expression of wild-type Socius in Swiss 3T3 fibroblasts has little effect on the actin cytoskeleton, the expression of a membrane-targeted form of Socius, containing a COOH-terminal farnesylation motif (Socius-CAAX), induces a dramatic loss of stress fibers. The inhibitory effect of Socius-CAAX on stress fiber formation is enhanced by truncation of its NH(2) terminus. On the other hand, the expression of Socius-CAAX or its NH(2) terminus-truncated form suppresses the Rnd induced retraction of the cell body and the production of extensively branching cellular processes, although the disassembly of stress fibers is observed. We propose that Socius participates in the Rnd GTPase-induced signal transduction pathways, leading to reorganization of the actin cytoskeleton. PMID- 11940654 TI - Counterregulation of chromatin deacetylation and histone deacetylase occupancy at the integrated promoter of human immunodeficiency virus type 1 (HIV-1) by the HIV 1 repressor YY1 and HIV-1 activator Tat. AB - Repression of human immunodeficiency virus type 1 (HIV-1) transcription may contribute to the establishment or maintenance of proviral quiescence in infected CD4(+) cells. The host factors YY1 and LSF cooperatively recruit histone deacetylase 1 (HDAC1) to the HIV-1 long terminal repeat (LTR) and inhibit transcription. We demonstrate here regulation of occupancy of HDAC1 at a positioned nucleosome (nuc 1) near the transcription start site of integrated LTR. We find that expression of YY1 increases occupancy by HDAC1, decreases acetylation at nuc 1, and downregulates LTR expression. HDAC1 recruitment and histone hypoacetylation were also seen when Tat activation was inhibited by the overexpression of YY1. A YY1 mutant without an HDAC1 interaction domain and incompetent to inhibit LTR activation fails to recruit HDAC1 to LTR or decrease nuc 1 acetylation. Further, expression of a dominant-negative mutant of LSF (dnLSF), which inhibits LSF occupancy and LTR repression, results in acetylation and decreased HDAC1 occupancy at nuc 1. Conversely, exposure of cells to the histone deacetylase inhibitor trichostatin A or activation of LTR expression by HIV-1 Tat results in the displacement of HDAC1 from nuc 1, in association with increased acetylation of histone H4. Recruitment of HDAC1 to the LTR nuc 1 can counteract Tat activation and repress LTR expression. Significantly, when repression is overcome, LTR activation is associated with decreased HDAC1 occupancy. Since the persistence of integrated HIV-1 genomes despite potent suppression of viral replication is a major obstacle for current antiretroviral therapy, strategies to selectively disrupt the quiescence of chromosomal provirus may play a role in the future treatment of AIDS. PMID- 11940655 TI - Dual roles of p300 in chromatin assembly and transcriptional activation in cooperation with nucleosome assembly protein 1 in vitro. AB - In a yeast two-hybrid screen to identify proteins that bind to the KIX domain of the coactivator p300, we obtained cDNAs encoding nucleosome assembly protein 1 (NAP-1), a 60-kDa histone H2A-H2B shuttling protein that promotes histone deposition. p300 associates preferentially with the H2A-H2B-bound form of NAP-1 rather than with the unbound form of NAP-1. Formation of NAP-1-p300 complexes was found to increase during S phase, suggesting a potential role for p300 in chromatin assembly. In micrococcal nuclease and supercoiling assays, addition of p300 promoted efficient chromatin assembly in vitro in conjunction with NAP-1 and ATP-utilizing chromatin assembly and remodeling factor; this effect was dependent in part on the intrinsic histone acetyltransferase activity of p300. Surprisingly, NAP-1 potently inhibited acetylation of core histones by p300, suggesting that efficient assembly requires acetylation of either NAP-1 or p300 itself. As p300 acted cooperatively with NAP-1 in stimulating transcription from a chromatin template in vitro, our results suggest a dual role of NAP-1-p300 complexes in promoting chromatin assembly and transcriptional activation. PMID- 11940656 TI - Carboxyl-terminal transactivation activity of hypoxia-inducible factor 1 alpha is governed by a von Hippel-Lindau protein-independent, hydroxylation-regulated association with p300/CBP. AB - Hypoxia-inducible factor 1 complex (HIF-1) plays a pivotal role in oxygen homeostasis and adaptation to hypoxia. Its function is controlled by both the protein stability and the transactivation activity of its alpha subunit, HIF-1 alpha. Hydroxylation of at least two prolyl residues in the oxygen-dependent degradation domain of HIF-1 alpha regulates its interaction with the von Hippel Lindau protein (VHL) that targets HIF-1 alpha for ubiquitination and proteasomal degradation. Several prolyl hydroxylases have been found to specifically hydroxylate HIF-1 alpha. In this report, we investigated possible roles of VHL and hydroxylases in the regulation of the transactivation activity of the C terminal activating domain (CAD) of HIF-1 alpha. We demonstrate that regulation of the transactivation activity of HIF-1 alpha CAD also involves hydroxylase activity but does not require functional VHL. In addition, stimulation of the CAD activity by a hydroxylase inhibitor, hypoxia, and desferrioxamine was severely blocked by the adenoviral oncoprotein E1A but not by an E1A mutant defective in targeting p300/CBP. We further demonstrate that a hydroxylase inhibitor, hypoxia, and desferrioxamine promote the functional and physical interaction between HIF-1 alpha CAD and p300/CBP in vivo. Taken together, our data provide evidence that hypoxia-regulated stabilization and transcriptional stimulation of HIF-1 alpha function are regulated through partially overlapping but distinguishable pathways. PMID- 11940657 TI - Altered p27(Kip1) phosphorylation, localization, and function in human epithelial cells resistant to transforming growth factor beta-mediated G(1) arrest. AB - p27(Kip1) is an important effector of G(1) arrest by transforming growth factor beta (TGF-beta). Investigations in a human mammary epithelial cell (HMEC) model, including cells that are sensitive (184(S)) and resistant (184A1L5(R)) to G(1) arrest by TGF-beta, revealed aberrant p27 regulation in the resistant cells. Cyclin E1-cyclin-dependent kinase 2 (cdk2) and cyclin A-cdk2 activities were increased, and p27-associated kinase activity was detected in 184A1L5(R) cells. p27 from 184A1L5(R) cells was localized to both nucleus and cytoplasm, showed an altered profile of phosphoisoforms, and had a reduced ability to bind and inhibit cyclin E1-cdk2 in vitro when compared to p27 from the sensitive 184(S) cells. In proliferating 184A1L5(R) cells, more p27 was associated with cyclin D1-cdk4 complexes than in 184(S). While TGF-beta inhibited the formation of cyclin D1 cdk4-p27 complexes in 184(S) cells, it did not inhibit the assembly of cyclin D1 cdk4-p27 complexes in the resistant 184A1L5(R) cells. p27 phosphorylation changed during cell cycle progression, with cyclin E1-bound p27 in G(0) showing a different phosphorylation pattern from that of cyclin D1-bound p27 in mid-G(1). These data suggest a model in which TGF-beta modulates p27 phosphorylation from its cyclin D1-bound assembly phosphoform to an alternate form that binds tightly to inhibit cyclin E1-cdk2. Altered phosphorylation of p27 in the resistant 184A1L5(R) cells may favor the binding of p27 to cyclin D1-cdk4 and prevent its accumulation in cyclin E1-cdk2 in response to TGF-beta. PMID- 11940658 TI - Calpain-mediated Bid cleavage and calpain-independent Bak modulation: two separate pathways in cisplatin-induced apoptosis. AB - Calpain is a ubiquitous protease with potential involvement in apoptosis. We report that in human melanoma cells, cisplatin-induced calpain activation occurs early in apoptosis. Calpain activation and subsequent apoptosis were inhibited by calpeptin and PD150606, two calpain inhibitors with different modes of action. Furthermore, cisplatin induced cleavage of the BH3-only protein Bid, yielding a 14-kDa fragment similar to proapoptotic, caspase-cleaved Bid. However, Bid cleavage was inhibited by inhibitors of calpain, but not by inhibitors of caspases or of cathepsin L. Recombinant Bid was cleaved in vitro by both recombinant calpain and by lysates of cisplatin-treated cells. Cleavage was calpeptin sensitive, and the cleavage site was mapped between Gly70 and Arg71. Calpain-cleaved Bid induced cytochrome c release from isolated mitochondria. While calpeptin did not affect cisplatin-induced modulation of Bak to its proapoptotic conformation, a dominant-negative mutant of MEKK1 (dnMEKK) inhibited Bak modulation. dnMEKK did not, however, block Bid cleavage. The combination of dnMEKK and calpeptin had an additive inhibitory effect on apoptosis. In summary, calpain-mediated Bid cleavage is important in drug-induced apoptosis, and cisplatin induces at least two separate apoptotic signaling pathways resulting in Bid cleavage and Bak modulation, respectively. PMID- 11940659 TI - MAD1 and p27(KIP1) cooperate to promote terminal differentiation of granulocytes and to inhibit Myc expression and cyclin E-CDK2 activity. AB - To understand how cellular differentiation is coupled to withdrawal from the cell cycle, we have focused on two negative regulators of the cell cycle, the MYC antagonist MAD1 and the cyclin-dependent kinase inhibitor p27(KIP1). Generation of Mad1/p27(KIP1) double-null mice revealed a number of synthetic effects between the null alleles of Mad1 and p27(KIP1), including embryonic lethality, increased proliferation, and impaired differentiation of granulocyte precursors. Furthermore, with granulocyte cell lines derived from the Mad1/p27(KIP1) double null mice, we observed constitutive Myc expression and cyclin E-CDK2 kinase activity as well as impaired differentiation following treatment with an inducer of differentiation. By contrast, similar treatment of granulocytes from Mad1 or p27(KIP1) single-null mice resulted in differentiation accompanied by downregulation of both Myc expression and cyclin E-CDK2 kinase activity. In the double-null granulocytic cells, addition of a CDK2 inhibitor in the presence of differentiation inducer was sufficient to restore differentiation and reduce Myc levels. We conclude that Mad1 and p27(KIP1) operate, at least in part, by distinct mechanisms to downregulate CDK2 activity and Myc expression in order to promote cell cycle exit during differentiation. PMID- 11940660 TI - Functional requirement for histone deacetylase 1 in Caenorhabditis elegans gonadogenesis. AB - Histone acetylation and deacetylation have been implicated in the regulation of gene expression. Molecular studies have shown that histone deacetylases (HDACs) function as transcriptional repressors. However, very little is known about their roles during development in multicellular organisms. We previously demonstrated that inhibition of maternal and zygotic expression of histone deacetylase 1 (HDA 1) causes embryonic lethality in Caenorhabditis elegans. Here, we report the identification of an hda-1 genetic mutant which has also been called a gon-10 mutant (for gonadogenesis defective 10) and show that loss of HDA-1 zygotic expression results in specific postembryonic defects in gonadogenesis and vulval development. We provide evidence that the lag-2 gene, which plays a role in gonadogenesis and vulval development and encodes a Notch ligand, is derepressed in gon-10 animals, suggesting that lag-2 may be a target of HDA-1. Our findings reveal a novel and specific function for the ubiquitously expressed HDA-1 in C. elegans gonadogenesis and place hda-1 in the Notch signaling pathway. PMID- 11940662 TI - Asthenozoospermia in mice with targeted deletion of the sperm mitochondrion associated cysteine-rich protein (Smcp) gene. AB - The sperm mitochondria-associated cysteine-rich protein (SMCP) is a cysteine- and proline-rich structural protein that is closely associated with the keratinous capsules of sperm mitochondria in the mitochondrial sheath surrounding the outer dense fibers and axoneme. To investigate the function of SMCP, we generated mice with a targeted disruption of the gene Smcp by homologous recombination. Homozygous mutant males on a mixed genetic background (C57BL/6J x 129/Sv) are fully fertile, while they are infertile on the 129/Sv background, although spermatogenesis and mating are normal. Homozygous Smcp(-/-) female mice are fertile on both genetic backgrounds. Electron microscopical examination demonstrated normal structures of sperm head, mitochondria, and tail. In vivo experiments with sperm of Smcp(-/-) 129/Sv mice revealed that the migration of spermatozoa from the uterus into the oviduct is reduced. This result is supported by the observation that sperm motility as determined by the computer-assisted semen analysis system (CASA) is significantly affected as compared to wild-type spermatozoa. In vitro fertilization assays showed that Smcp-deficient spermatozoa are able to bind to the oocyte but that the number of fertilized eggs is reduced by more than threefold relative to the wild-type control. However, removal of the zona pellucida resulted in an unaffected sperm-egg fusion which was monitored by the presence of pronuclei and generation of blastocyts. These results indicate that the infertility of the male Smcp(-/-) mice on the 129/Sv background is due to reduced motility of the spermatozoa and decreased capability of the spermatozoa to penetrate oocytes. PMID- 11940661 TI - Kinase suppressor of Ras (KSR) is a scaffold which facilitates mitogen-activated protein kinase activation in vivo. AB - While scaffold proteins are thought to be key components of signaling pathways, their exact function is unknown. By preassembling multiple components of signaling cascades, scaffolds are predicted to influence the efficiency and/or specificity of signaling events. Here we analyze a potential scaffold of the Ras/mitogen-activated protein kinase (MAPK) pathway, kinase suppressor of Ras (KSR), by generating KSR-deficient mice. KSR-deficient mice were grossly normal even though ERK kinase activation was attenuated to a degree sufficient to block T-cell activation and inhibit tumor development. Consistent with its role as a scaffold, high-molecular-weight complexes containing KSR, MEK, and ERK were lost in the absence of KSR. This demonstrates that KSR is a bona fide scaffold that is not required for but enhances signaling via the Ras/MAPK signaling pathway. PMID- 11940663 TI - Initiation sites are distributed at frequent intervals in the Chinese hamster dihydrofolate reductase origin of replication but are used with very different efficiencies. AB - Previous radiolabeling and two-dimensional (2-D) gel studies of the dihydrofolate reductase (DHFR) domain of Chinese hamster cells have suggested that replication can initiate at any one of a very large number of inefficient sites scattered throughout the 55-kb intergenic spacer region, with two broad subregions (ori beta and ori-gamma) preferred. However, high-resolution analysis by a PCR-based nascent strand abundance assay of the 12-kb subregion encompassing ori-beta has suggested the presence of a relatively small number of fixed, highly efficient initiation sites distributed at infrequent intervals that correspond to genetic replicators. To attempt to reconcile these observations, two different approaches were taken in the present study. In the first, neutral-neutral 2-D gel analysis was used to examine replication intermediates in 31 adjacent and overlapping restriction fragments in the spacer, ranging in size from 1.0 to 18 kb. Thirty of 31 fragments displayed the complete bubble arcs characteristic of centered origins. Taking into account overlapping fragments, these data suggest a minimum of 14 individual start sites in the spacer. In the second approach, a quantitative early labeled fragment hybridization assay was performed in which radioactive origin-containing DNA 300 to 1,000 nucleotides in length was synthesized in the first few minutes of the S period and used to probe 15 clones distributed throughout the intergenic spacer but separated on average by more than 1,000 bp. This small nascent DNA fraction hybridized to 14 of the 15 clones, ranging from just above background to a maximum at the ori-beta locus. The only silent region detected was a small fragment lying just upstream from a centered matrix attachment region--the same region that was also negative for initiation by 2-D gel analysis. Results of both approaches suggest a minimum of approximately 20 initiation sites in the spacer (two of them being ori-beta and ori-gamma), with ori-beta accounting for a maximum of approximately 20% of initiations occurring in the spacer. We believe that the results of all experimental approaches applied to this locus so far can be fitted to a model in which the DHFR origin consists of a 55-kb intergenic zone of potential sites that are used with very different efficiencies and which are separated in many cases by a few kilobases or less. PMID- 11940664 TI - SH2-B is required for both male and female reproduction. AB - Many growth factors and hormones modulate the reproductive status in mammals. Among these, insulin and insulin-like growth factor I (IGF-I) regulate the development of gonadal tissues. SH2-B has been shown to interact with insulin and IGF-I receptors, although the role of SH2-B in these signals has not been clarified. To investigate the role of SH2-B, we generated mice with a targeted disruption of the SH2-B gene. Both male and female SH2-B(-/-) mice showed slight retardation in growth and impaired fertility. Female knockout mice possess small, anovulatory ovaries with reduced numbers of follicles and male SH2-B(-/-) mice have small testes with a reduced number of sperm. SH2-B(-/-) cumulus cells do not respond to either follicle-stimulating hormone or IGF-I. These data suggest that SH2-B plays a critical role in the IGF-I-mediated reproductive pathway in mice. PMID- 11940665 TI - The Mnd1 protein forms a complex with hop2 to promote homologous chromosome pairing and meiotic double-strand break repair. AB - The hop2 mutant of Saccharomyces cerevisiae arrests in meiosis with extensive synaptonemal complex (SC) formation between nonhomologous chromosomes. A screen for multicopy suppressors of a hop2-ts allele identified the MND1 gene. The mnd1 null mutant arrests in meiotic prophase, with most double-strand breaks (DSBs) unrepaired. A low level of mature recombinants is produced, and the Rad51 protein accumulates at numerous foci along chromosomes. SC formation is incomplete, and homolog pairing is severely reduced. The Mnd1 protein localizes to chromatin throughout meiotic prophase, and this localization requires Hop2. Unlike recombination enzymes such as Rad51, Mnd1 localizes to chromosomes even in mutants that fail to initiate meiotic recombination. The Hop2 and Mnd1 proteins coimmunoprecipitate from meiotic cell extracts. These results suggest that Hop2 and Mnd1 work as a complex to promote meiotic chromosome pairing and DSB repair. The identification of Hop2 and Mnd1 homologs in other organisms suggests that the function of this complex is conserved among eukaryotes. PMID- 11940666 TI - LIS1, CLIP-170's key to the dynein/dynactin pathway. AB - CLIP-170 is a plus-end tracking protein which may act as an anticatastrophe factor. It has been proposed to mediate the association of dynein/dynactin to microtubule (MT) plus ends, and it also binds to kinetochores in a dynein/dynactin-dependent fashion, both via its C-terminal domain. This domain contains two zinc finger motifs (proximal and distal), which are hypothesized to mediate protein-protein interactions. LIS1, a protein implicated in brain development, acts in several processes mediated by the dynein/dynactin pathway by interacting with dynein and other proteins. Here we demonstrate colocalization and direct interaction between CLIP-170 and LIS1. In mammalian cells, LIS1 recruitment to kinetochores is dynein/dynactin dependent, and recruitment there of CLIP-170 is dependent on its site of binding to LIS1, located in the distal zinc finger motif. Overexpression of CLIP-170 results in a zinc finger-dependent localization of a phospho-LIS1 isoform and dynactin to MT bundles, raising the possibility that CLIP-170 and LIS1 regulate dynein/dynactin binding to MTs. This work suggests that LIS1 is a regulated adapter between CLIP-170 and cytoplasmic dynein at sites involved in cargo-MT loading, and/or in the control of MT dynamics. PMID- 11940667 TI - Tumor suppression by a severely truncated species of retinoblastoma protein. AB - Rb(+/+):Rb(-/-) chimeric mice are healthy until early in adulthood when they develop lethal pituitary tumors composed solely of Rb(-/-) cells. In an effort to delineate the minimal structures of the retinoblastoma protein necessary for RB tumor suppression function, chimeric animals derived from stably transfected RB( /-) embryonic stem (ES) cells were generated. One such ES cell transfectant expressed a human RB allele encoding a stable, truncated nuclear derivative lacking residues 1 to 378 (Delta 1-378). Others encoded either wild-type human RB or an internally deleted derivative of the Delta 1-378 mutant. All gave rise to viable chimeric animals with comparable degrees of chimerism. However, unlike control mice derived, in part, from naive Rb(-/-) ES cells or from ES cells transformed by the double RB mutant, Delta 1-378/Delta exon22, animals derived from either wild-type RB- or Delta 1-378 RB-producing ES cells failed to develop pituitary tumors. Thus, in this setting, a substantial fraction of the RB sequence is unnecessary for RB-mediated tumor suppression. PMID- 11940668 TI - Conserved furin cleavage site not essential for secretion and integration of ZP3 into the extracellular egg coat of transgenic mice. AB - The extracellular zona pellucida surrounding mammalian eggs is formed by interactions of the ZP1, ZP2, and ZP3 glycoproteins. Female mice lacking ZP2 or ZP3 do not form a stable zona matrix and are sterile. The three zona proteins are synthesized in growing oocytes and secreted prior to incorporation into the zona pellucida. A well-conserved furin site upstream of a transmembrane domain near the carboxyl terminus of each has been implicated in the release of the zona ectodomains from oocytes. However, mutation of the furin site (RNRR --> ANAA) does not affect the intracellular trafficking or secretion of an enhanced green fluorescent protein (EGFP)-ZP3 fusion protein in heterologous somatic cells. After transient expression in growing oocytes, normal EGFP-ZP3 and mutant EGFP ZP3 associate with the inner aspect of the zona pellucida, which is distinct from the plasma membrane. These in vitro results are confirmed in transgenic mice expressing EGFP-ZP3 with or without the mutant furin site. In each case, EGFP-ZP3 is incorporated throughout the width of the zona pellucida and the transgenic mice are fertile. These results indicate that the zona matrix accrues from the inside out and, unexpectedly, suggest that cleavage at the furin site is not required for formation of the extracellular zona pellucida surrounding mouse eggs. PMID- 11940669 TI - Interaction between FOG-1 and the corepressor C-terminal binding protein is dispensable for normal erythropoiesis in vivo. AB - The hematopoietic, zinc-finger protein FOG-1 is essential for the development of the erythroid and megakaryocytic lineages. FOG-1's function in hematopoiesis is dependent on its ability to interact with the transcription factor GATA-1. FOG-1 has also been observed to interact with the corepressor molecule C-terminal binding protein (CtBP) through a peptide motif shared by all FOG family members. In this study, we confirmed that FOG-1 and CtBP interact by coimmunoprecipitation. We further demonstrate that a FOG-1 mutant unable to interact with CtBP has increased erythropoietic (but not megakaryocytic) rescue (relative to the wild type) of a FOG-1(-/-) cell line. To analyze further the physiological role of the FOG-1-CtBP interaction, we generated knock-in mice that express a FOG-1 variant unable to bind CtBP. Surprisingly, these mice are normal and fertile. Furthermore, erythropoiesis at all stages of development is normal in these mice. Erythrocyte production is similar in mutant and wild-type mice even under conditions of erythropoietic stress stimulated by either exogenously added erythropoietin or phenylhydrazine-induced anemia. Thus, despite conservation of the FOG-CtBP interaction site, the in vivo function of FOG-1 in erythroid development is not affected by its inability to interact with the corepressor CtBP. PMID- 11940670 TI - Notch signaling induces rapid degradation of achaete-scute homolog 1. AB - In neural development, Notch signaling plays a key role in restricting neuronal differentiation, promoting the maintenance of progenitor cells. Classically, Notch signaling causes transactivation of Hairy-enhancer of Split (HES) genes which leads to transcriptional repression of neural determination and differentiation genes. We now report that in addition to its known transcriptional mechanism, Notch signaling also leads to rapid degradation of the basic helix-loop-helix (bHLH) transcription factor human achaete-scute homolog 1 (hASH1). Using recombinant adenoviruses expressing active Notch1 in small-cell lung cancer cells, we showed that the initial appearance of Notch1 coincided with the loss of hASH1 protein, preceding the full decay of hASH1 mRNA. Overexpression of HES1 alone was capable of down-regulating hASH1 mRNA but could not replicate the acute reduction of hASH1 protein induced by Notch1. When adenoviral hASH1 was coinfected with Notch1, we still observed a dramatic and abrupt loss of the exogenous hASH1 protein, despite high levels of ongoing hASH1 RNA expression. Notch1 treatment decreased the apparent half-life of the adenoviral hASH1 protein and increased the fraction of hASH1 which was polyubiquitinylated. The proteasome inhibitor MG132 reversed the Notch1-induced degradation. The Notch RAM domain was dispensable but a lack of the OPA and PEST domains inactivated this Notch1 action. Overexpression of the hASH1-dimerizing partner E12 could protect hASH1 from degradation. This novel function of activated Notch to rapidly degrade a class II bHLH protein may prove to be important in many contexts in development and in cancer. PMID- 11940671 TI - Defective dendrite elongation but normal fertility in mice lacking the Rho-like GTPase activator Dbl. AB - Dbl is the prototype of a large family of GDP-GTP exchange factors for small GTPases of the Rho family. In vitro, Dbl is known to activate Rho and Cdc42 and to induce a transformed phenotype. Dbl is specifically expressed in brain and gonads, but its in vivo functions are largely unknown. To assess its role in neurogenesis and gametogenesis, targeted deletion of the murine Dbl gene was accomplished in embryonic stem cells. Dbl-null mice are viable and did not show either decreased reproductive performances or obvious neurological defects. Histological analysis of mutant testis showed normal morphology and unaltered proliferation and survival of spermatogonia. Dbl-null brains indicated a correct disposition of the major neural structures. Analysis of cortical stratification indicated that Dbl is not crucial for neuronal migration. However, in distinct populations of Dbl-null cortical pyramidal neurons, the length of dendrites was significantly reduced, suggesting a role for Dbl in dendrite elongation. PMID- 11940672 TI - Transcription factor gene AP-2 gamma essential for early murine development. AB - Transcription factor gene AP-2 gamma belongs to a family of four closely related genes. AP-2 gamma had been implicated in multiple functions during proliferation and differentiation based on its expression pattern in trophoblast, neural crest, and ectoderm cells in murine embryos. In order to address the question of the role of AP-2 gamma during mammalian development, we generated mice harboring a disrupted AP-2 gamma allele. AP-2 gamma heterozygous mice are viable and display reduced body sizes at birth but are fertile. Mice deficient for AP-2 gamma, however, are growth retarded and die at days 7 to 9 of embryonic development. Immunohistochemical analysis revealed that the trophectodermal cells that are found to express AP-2 gamma fail to proliferate, leading to failure of labyrinth layer formation. As a consequence, the developing embryo suffers from malnutrition and dies. Analysis of embryo cultures suggests that AP-2 gamma is also implicated in the regulation of the adenosine deaminase (ADA) gene, a gene involved in purine metabolism found expressed at the maternal-fetal interface. Therefore, AP-2 gamma seems to be required in early embryonic development because it regulates the genetic programs controlling proliferation and differentiation of extraembryonic trophectodermal cells. PMID- 11940673 TI - Molecular mechanisms of gene silencing mediated by DNA methylation. AB - DNA methylation and chromatin modification operate along a common pathway to repress transcription; accordingly, several experiments demonstrate that the effects of DNA methylation can spread in cis and do not require promoter modification. In order to investigate the molecular details of the inhibitory effect of methylation, we microinjected into Xenopus oocytes a series of constructs containing a human CpG-rich sequence which has been differentially methylated and cloned at different positions relative to a specific promoter. The parameters influencing the diffusion of gene silencing and the importance of histone deacetylation in the spreading effect were analyzed. We demonstrate that a few methylated cytosines can inhibit a flanking promoter but a threshold of modified sites is required to organize a stable, diffusible chromatin structure. Histone deacetylation is the main cause of gene repression only when methylation does not reach levels sufficient to establish this particular structure. Moreover, contrary to the common thought, promoter modification does not lead to the greater repressive effect; the existence of a competition between transactivators and methyl-binding proteins for the establishment of an open conformation justifies the results obtained. PMID- 11940674 TI - RAG-mediated V(D)J recombination is not essential for tumorigenesis in Atm deficient mice. AB - Atm-deficient mice die of malignant thymic lymphomas characterized by translocations within the Tcr alpha/delta locus, suggesting that tumorigenesis is secondary to aberrant responses to double-stranded DNA (dsDNA) breaks that occur during RAG-dependent V(D)J recombination. We recently demonstrated that development of thymic lymphoma in Atm(-/-) mice was not prevented by loss of RAG 2. Thymic lymphomas that developed in Rag2(-/-) Atm(-/-) mice contained multiple chromosomal abnormalities, but none of these involved the Tcr alpha/delta locus. These findings indicated that tumorigenesis in Atm(-/-) mice is mediated by chromosomal translocations secondary to aberrant responses to dsDNA breaks and that V(D)J recombination is an important, but not essential, event in susceptibility. In contrast to these findings, it was recently reported that Rag1(-/-) Atm(-/-) mice do not develop thymic lymphomas, a finding that was interpreted as demonstrating a requirement for RAG-dependent recombination in the susceptibility to tumors in Atm-deficient mice. To test the possibility that RAG 1 and RAG-2 differ in their roles in tumorigenesis, we studied Rag1(-/-) Atm(-/-) mice in parallel to our previous Rag2(-/-) Atm(-/-) study. We found that thymic lymphomas occur at high frequency in Rag1(-/-) Atm(-/-) mice and resemble those that occur in Rag2(-/-) Atm(-/-) mice. These results indicate that both RAG-1 and RAG-2 are necessary for tumorigenesis involving translocation in the Tcr alpha/delta locus but that Atm deficiency leads to tumors through a broader RAG independent predisposition to translocation, related to a generalized defect in dsDNA break repair. PMID- 11940675 TI - Distinct mutations in yeast TAF(II)25 differentially affect the composition of TFIID and SAGA complexes as well as global gene expression patterns. AB - The RNA polymerase II transcription factor TFIID, composed of the TATA-binding protein (TBP) and TBP-associated factors (TAF(II)s), nucleates preinitiation complex formation at protein-coding gene promoters. SAGA, a second TAF(II) containing multiprotein complex, is involved in transcription regulation in Saccharomyces cerevisiae. One of the essential protein components common to SAGA and TFIID is yTAF(II)25. We define a minimal evolutionarily conserved 91-amino acid region of TAF(II)25 containing a histone fold domain that is necessary and sufficient for growth in vivo. Different temperature-sensitive mutations of yTAF(II)25 or chimeras with the human homologue TAF(II)30 arrested cell growth at either the G(1) or G(2)/M cell cycle phase and displayed distinct phenotypic changes and gene expression patterns. Immunoprecipitation studies revealed that TAF(II)25 mutation-dependent gene expression and phenotypic changes correlated at least partially with the integrity of SAGA and TFIID. Genome-wide expression analysis revealed that the five TAF(II)25 temperature-sensitive mutant alleles individually affect the expression of between 18 and 33% of genes, whereas taken together they affect 64% of all class II genes. Thus, different yTAF(II)25 mutations induce distinct phenotypes and affect the regulation of different subsets of genes, demonstrating that no individual TAF(II) mutant allele reflects the full range of its normal functions. PMID- 11940678 TI - Dementia, butterfly ballots, and voter competence. PMID- 11940676 TI - Assembly and function of the RNA editing complex in Trypanosoma brucei requires band III protein. AB - Trypanosome RNA editing, the posttranscriptional insertion and deletion of U residues in mitochondrial transcripts, is catalyzed by a protein complex containing seven distinct proteins. In this study, we cloned the gene for band III, a 555-amino-acid protein with two separate zinc finger motifs. We prepared antibodies that showed band III protein cofractionates with the previously characterized band IV protein throughout the purification of the editing complex and is not found free or in other protein associations; therefore, it is a true constituent of the editing complex. Double-stranded RNA interference efficiently depleted band III protein and demonstrated that band III expression is essential for growth of procyclic trypanosomes and for RNA editing. These depleted cell extracts were deficient specifically in guide RNA-directed endonuclease cleavage at both U deletion and U insertion sites and in the activity of the band IV ligase, but they retained the 3'-U-exonuclease and terminal-U-transferase activities as well as band V ligase of the editing complex. Loss of band III protein also resulted in almost complete loss of the band IV ligase protein and altered sedimentation of the band V ligase. These data indicate that band III is either the RNA editing endonuclease or a factor critical for cleavage activity in the editing complex. They also demonstrate that band III is required for proper assembly of the editing complex. PMID- 11940679 TI - Can imaging distinguish PSP from other neurodegenerative disorders? PMID- 11940681 TI - Lawrence D. Jacobs, MD (1938-2001). PMID- 11940677 TI - Heterochromatin protein 1 is involved in control of telomere elongation in Drosophila melanogaster. AB - Telomeres of Drosophila melanogaster contain arrays of the retrotransposon-like elements HeT-A and TART. Their transposition to broken chromosome ends has been implicated in chromosome healing and telomere elongation. We have developed a genetic system which enables the determination of the frequency of telomere elongation events and their mechanism. The frequency differs among lines with different genotypes, suggesting that several genes are in control. Here we show that the Su(var)2-5 gene encoding heterochromatin protein 1 (HP1) is involved in regulation of telomere length. Different Su(var)2-5 mutations in the heterozygous state increase the frequency of HeT-A and TART attachment to the broken chromosome end by more than a hundred times. The attachment occurs through either HeT-A/TART transposition or recombination with other telomeres. Terminal DNA elongation by gene conversion is greatly enhanced by Su(var)2-5 mutations only if the template for DNA synthesis is on the same chromosome but not on the homologous chromosome. The Drosophila lines bearing the Su(var)2-5 mutations maintain extremely long telomeres consisting of HeT-A and TART for many generations. Thus, HP1 plays an important role in the control of telomere elongation in D. melanogaster. PMID- 11940682 TI - Familial transthyretin-type amyloid polyneuropathy in Japan: clinical and genetic heterogeneity. AB - Familial amyloid polyneuropathy (FAP) was once considered a disease peculiar to endemic areas, but it is now recognized not to be a rare disease among hereditary neuropathic disorders in Japan. FAP in Japan, the majority of which is caused by transthyretin (TTR)-related amyloid deposition, shows a wide spectrum of clinical pictures. This variability can be explained on the basis of the many causative gene mutations of TTR, but even in the same TTR type of FAP, the clinical phenotypes seem to vary in different kindreds or individuals. Especially in the case of the Val30Met TTR type, the sex ratio and the age at onset are considerably different between patients in endemic foci and those in nonendemic areas. It is also noteworthy that serious cardiac amyloidosis is commonly seen in patients with FAP of the non-Val30Met TTR type. In addition to TTR gene mutation, unknown factors may play an important role in the development of FAP. At present, liver transplantation is the only life-saving treatment, but this therapy is always associated with great stress for the patient and the donor, particularly in living-related transplantation. Less invasive treatments for this disease are required. PMID- 11940683 TI - Morbidity and mortality following pallidotomy in Parkinson's disease: a systematic review. AB - OBJECTIVE: To study the frequency of morbidity and mortality associated with pallidotomy. METHOD: The authors searched the MEDLINE electronic database for pallidotomy articles reported between January 1992 and December 2000. They selected studies by the following criteria: original clinical data, unequivocal description of morbidity and mortality, and unselected consecutive cases. The authors extracted the following data: number of patients, unilateral or bilateral procedures, age, localization technique, follow-up time, number of patients with adverse effects, number of patients with permanent adverse effects (>3 months), types of adverse effects, and mortality. RESULTS: For unilateral pallidotomy, 12 prospective studies included 334 patients. Of these patients, 30.2% (95% CI, 25.3 to 35.2) had adverse effects, and 13.8% (95% CI, 10.1 to 17.5) had permanent adverse effects. A symptomatic infarction or hemorrhage occurred in 3.9% (95% CI, 2.1 to 6.6). The mortality rate was 1.2% (95% CI, 0.3 to 3.0). In the series with microelectrode recording, the frequency of adverse effects was 14.4% (95% CI, 4.7 to 24.1) higher and the frequency of stroke was 4.9% (95% CI, 1.4 to 8.4) higher. The most frequent adverse effects were problems with speech (11.1%) and facial paresis (8.4%). For bilateral pallidotomy, five historical cohort studies including 20 patients were available for review. Fourteen patients had an adverse effect, and the most frequent adverse effects were impairments of speech and cognition. CONCLUSIONS: The risk of permanent adverse effects associated with unilateral pallidotomy was 13.8%. A symptomatic infarction or hemorrhage occurred in 3.9% of patients, and the associated mortality rate was 1.2%. PMID- 11940684 TI - Cholinergic vesicular transporters in progressive supranuclear palsy. AB - OBJECTIVE: To determine the status of cholinergic and monoaminergic vesicular transporter binding sites in progressive supranuclear palsy (PSP). METHODS: The authors determined autoradiographically the regional expression of acetylcholine vesicular transporter (VAChT) and monoamine vesicular transporter type 2 (VMAT2) binding sites in postmortem basal ganglia samples from subjects with PSP. Comparison neurochemical measures included choline acetyltransferase (ChAT) enzyme activity and benzodiazepine (BZ) binding sites. RESULTS: VAChT expressions and ChAT activities in caudate nucleus and putamen were markedly decreased in PSP, whereas BZ binding was unaffected, consistent with selective losses of striatal cholinergic interneurons. VMAT2 density was reduced significantly in the caudate nucleus, putamen, and substantia nigra pars compacta, consistent with degeneration of dopaminergic nigrostriatal projection neurons in PSP. In the globus pallidus, BZ receptor binding sites were reduced, whereas VMAT2 and VAChT binding sites were unchanged, indicating losses of intrinsic pallidal neurons and synapses. CONCLUSIONS: These results confirm selective and marked degenerations of basal ganglia cholinergic and dopaminergic terminals in PSP. Striatal VAChT reduction may provide a unique neurochemical imaging marker for distinction of PSP from other types of basal ganglia neurodegeneration. PMID- 11940685 TI - Parkinson's disease and sleepiness: an integral part of PD. AB - OBJECTIVE: To investigate the potential causes of excessive daytime sleepiness in patients with PD-poor sleep quality, abnormal sleep-wakefulness control, and treatment with dopaminergic agents. METHODS: The authors performed night-time polysomnography and daytime multiple sleep latency tests in 54 consecutive levodopa-treated patients with PD referred for sleepiness, 27 of whom were also receiving dopaminergic agonists. RESULTS: Sleep latency was 6.3 +/- 0.6 minutes (normal >8 minutes), and the Epworth Sleepiness score was 14.3 +/- 4.1 (normal <10). A narcolepsy-like phenotype (> or = 2 sleep-onset REM periods) was found in 39% of the patients, who were sleepier (4.6 +/- 0.9 minutes) than the other 61% of patients (7.4 +/- 0.7 minutes). Periodic leg movement syndromes were rare (15%, range 16 to 43/h), but obstructive sleep apnea-hypopnea syndromes were frequent (20% of patients had an apnea-hypopnea index >15/h; range 15.1 to 50.0). Severity of sleepiness was weakly correlated with Epworth Sleepiness score (r = 0.34) and daily dose of levodopa (r = 0.30) but not with dopamine-agonist treatment, age, disease duration, parkinsonian motor disability, total sleep time, periodic leg movement, apnea-hypopnea, or arousal indices. CONCLUSIONS: In patients with PD preselected for sleepiness, severity of sleepiness was not dependent on nocturnal sleep abnormalities, motor and cognitive impairment, or antiparkinsonian treatment. The results suggest that sleepiness-sudden onset of sleep-does not result from pharmacotherapy but is related to the pathology of PD. PMID- 11940687 TI - Japanese family with parkinsonism, depression, weight loss, and central hypoventilation. AB - BACKGROUND: The authors describe the clinical and pathologic characteristics of the Fukuoka 1 family, the first Japanese family recognized to have hereditary parkinsonism associated with depression, weight loss, and central alveolar hypoventilation. METHODS: The pedigree contains 14 family members spanning four generations, with five affected individuals. All available medical records were collected for affected members, including autopsy results. RESULTS: The inheritance pattern was autosomal dominant. The average age at onset of symptoms was 41 years. All patients had parkinsonism characterized by rigidity, bradykinesia, and resting and postural tremor. Bradykinesia and depression developed in the proband at age 43 years. He responded to levodopa in the initial stage only. A year later, he had weight loss and central hypoventilation leading to respiratory failure. Symptoms developed in his cousin at age 38 years. The proband's father developed a resting tremor and depression at age 43 years. The tremor was initially responsive to levodopa therapy, but the disease was relentlessly progressive, leading to severe bradykinesia, rigidity, weight loss, and respiratory distress. He died of respiratory failure at age 49 years. Autopsy showed marked neuronal loss and gliosis in the substantia nigra and locus ceruleus. Lewy bodies, neurofibrillary tangles, senile plaques, and other abnormal structures were not seen in the cortical and subcortical regions. CONCLUSIONS: The Fukuoka 1 family shares many clinical and pathologic features with five previously reported kindreds from North America and Europe, suggesting that this syndrome has a worldwide distribution and can occur in different ethnic populations. PMID- 11940688 TI - Autosomal dominant chorea-acanthocytosis with polyglutamine-containing neuronal inclusions. AB - BACKGROUND: The term chorea-acanthocytosis describes a heterogeneous group of neurodegenerative disorders with variable clinical features and modes of inheritance. The characteristic acanthocytic appearance of red blood cells is attributed to abnormalities of a membrane protein, band 3, although the relationship between this and the neurodegenerative process has yet to be determined. OBJECTIVE: To describe features of phenotype, inheritance, and neuropathological findings in a family with this disorder. METHODS: Clinical and hematologic evaluations were performed on all available family members and neuropathological examination was performed on one case. RESULTS: Autosomal dominant inheritance was evident, with variable clinical features of chorea or parkinsonism, marked cognitive changes, but no seizures or peripheral neurologic abnormalities. Abnormalities of band 3 were demonstrated on gel electrophoresis of red blood cell membranes. Neuropathological examination revealed severe neuronal loss of the caudate-putamen and intranuclear inclusion bodies in many areas of the cerebral cortex. These inclusion bodies were immunoreactive for ubiquitin, expanded polyglutamine repeats, and torsinA. CONCLUSIONS: This family extends the genetic spectrum of chorea-acanthocytosis to include autosomal dominant inheritance, possibly due to expanded trinucleotide repeats. Intraneuronal inclusion bodies have recently been associated with a wide range of inherited neurodegenerative disorders and may provide a clue to etiopathogenesis, in addition to potentially indicating a function of torsinA. PMID- 11940689 TI - Apolipoprotein E epsilon 4 and short-term recovery from predominantly mild brain injury. AB - OBJECTIVE: To determine whether APOE genotype explained variability in short-term recovery from predominantly mild traumatic brain injury (TBI). METHODS: A total of 87 adult patients presenting with mild or moderate TBI to a shock trauma center were enrolled prospectively. A battery of 13 neuropsychological tests was administered twice, at approximately 3 and 6 weeks after injury. Eighty of 87 patients were successfully genotyped for APOE using a buccal swab technique. RESULTS: Ninety percent of study patients had mild TBI (Glasgow Coma Scale score of 13 to 15); 18 (22.5%) had one APOE epsilon 4 and none had two epsilon 4 alleles. After adjusting for potential confounders, patients positive for the APOE epsilon 4 allele had lower mean scores on 12 of 13 neuropsychological outcomes at visit 1 compared with APOE epsilon 4-negative patients. Two of the differences were significant (grooved pegboard test, p = 0.005; paced auditory serial addition task 2.8-second trial, p = 0.004). At visit 2, APOE epsilon 4 positive patients had lower adjusted mean scores on 11 of the 13 neuropsychological outcomes. None of the differences was significant. CONCLUSIONS: APOE genotype may influence the severity of the acute injury. However, with no consistent pattern to the recovery curves, it is not clear if APOE genotype influences the rate of recovery. PMID- 11940690 TI - APOE genotype and survival in men and women with Alzheimer's disease. AB - BACKGROUND: The epsilon 4 allele of the APOE gene (APOE) is more frequent in patients with AD than in the general population, but studies are inconclusive as to whether it affects rate of progression or survival. Because survival in AD is generally longer in women than in men, the authors investigated whether APOE affects 10-year survival equally in men and women. METHODS: APOE testing was performed on 125 patients with probable AD enrolled in the Johns Hopkins AD Research Center between November 1984 and March 1987. The 39 men and 86 women were followed at 6-month intervals until censoring (by death or withdrawal from the study) or March 1997. Patients were dichotomized into those with and those without at least one epsilon 4 allele. For each sex, a Cox proportional hazards regression, allowing for delayed entry and covarying for age at onset, was used to examine the effect of epsilon 4 on survival. RESULTS: All patients who died during the study period and had autopsy (n = 92) were found to have definite AD. Average survival from disease onset did not differ by sex (12.1 years in men; 12.3 years in women). In neither sex were differences found between epsilon 4 positive and epsilon 4-negative subgroups in education, duration of AD at entry, or severity of dementia. However, in both sexes the epsilon 4-positive subgroup was approximately 3 years older at onset of AD and at entry to the study than the epsilon 4-negative subgroup. Adjusting for age at onset, the presence of an epsilon 4 allele significantly increased the relative risk of death only for men (RR = 2.69; 95% CI = 1.23 to 5.87). CONCLUSIONS: In this sample of mostly white, well-educated research participants with AD, the APOE epsilon 4 allele was associated with shorter survival in men but not in women. PMID- 11940691 TI - Randomized pilot study of nimesulide treatment in Alzheimer's disease. AB - BACKGROUND: Nonsteroidal anti-inflammatory drugs (NSAID) may be useful in the treatment of AD. Clinical and laboratory experience with nimesulide, an NSAID with preferential cyclooxygenase-2 inhibition, suggests that it may be a good candidate for AD therapy. METHODS: This pilot study investigated the clinical feasibility of nimesulide treatment in AD. Forty persons with probable AD, most of whom were taking cholinesterase inhibitors, were enrolled in a randomized, controlled, parallel-group trial designed to assess tolerability and short-term cognitive/behavioral effects of nimesulide. In the initial 12-week double-blind phase, participants were treated with nimesulide 100 mg by mouth twice daily or matching placebo; during the second 12-week phase all participants received active drug. Participants who tolerated the drug well and perceived benefit were invited to continue open-label nimesulide treatment. RESULTS: Short-term therapy with nimesulide, compared with placebo, had no significant effect on total assessment scores of measures of cognition, clinical status, activities of daily living, affect, and behavior. Long-term therapy was well tolerated for periods exceeding 2 years. CONCLUSION: These findings support the feasibility of nimesulide therapy in AD; assessment of efficacy will require a larger, long-term treatment study. PMID- 11940692 TI - Serum thyroxine level and cognitive decline in euthyroid older women. AB - BACKGROUND: Clinical and subclinical hypothyroidism is associated with cognitive impairment. OBJECTIVE: This study investigated the association between thyroxine (T(4)) and thyroid-stimulating hormone (TSH) level and change over time in cognitive performance in a sample of older women with normal thyroid gland function. METHODS: T(4) and TSH were measured at baseline in 628 women (> or = 65 years) enrolled in the Women's Health and Aging Study, a community-based study of physically impaired women. Cognitive function was assessed at baseline and after 1, 2, and 3 years, using the Mini-Mental State Examination (MMSE). Incident cognitive decline was defined as a decrease of more than one point/year in MMSE score between baseline and the end of the follow-up. The analysis included 464 subjects with normal thyroid gland function with a baseline and at least one follow-up MMSE. RESULTS: At baseline there was no association between T(4) and TSH level and cognitive function. In longitudinal analysis, adjusting for age, race, level of education, and other covariates, compared with women in the highest T(4) tertile (8.1 to 12.5 microg/dL), those in the lowest tertile (4.5 to 6.5 microg/dL) had a greater decline in MMSE score (-0.25 point/year vs -0.12 point/year; p = 0.04). A total of 95 women (20.5%) had cognitive decline during the study period (mean MMSE decline, 5.5 points). Compared with women in the highest T(4) tertile, those in the lowest tertile had a twofold risk of cognitive decline (adjusted relative risk, 1.97; 95% CI, 1.10 to 3.50). The results were not modified by baseline cognitive and physical function. There was no association between baseline TSH level and change in cognitive function. CONCLUSIONS: In older women, low T(4) levels, within the normal range, were associated with a greater risk of cognitive decline over a 3-year period. Thyroid hormone levels may contribute to cognitive impairment in physically impaired women. PMID- 11940693 TI - Quality of life in chronic daily headache: a study in a general population. AB - OBJECTIVE: To analyze the quality of life (QoL) of subjects with chronic daily headache (CDH) in the general population. METHODS: QoL was studied, using the generic instrument Short Form-36 (SF-36), in 89 unselected subjects from an epidemiologic study in the general population who fulfilled CDH criteria. SF-36 scores were adjusted for comorbid conditions. A total of 89 healthy matched subjects were recruited as a control group. An additional matched group of 89 otherwise healthy subjects with episodic migraine was recruited as controls only to those with transformed migraine (TM). RESULTS: CDH subjects showed a significant decrease in each health-related concept of the SF-36 as compared with healthy subjects. The highest decreases were seen for role physical, bodily pain, vitality, and social functioning. There was no significant difference in SF-36 scores in subjects with chronic tension-type headache as compared with TM subjects. TM individuals showed lower values in each health-related concept when compared with patients with episodic migraine, these decreases being significant for general health, vitality, and mental health. Finally, CDH subjects without analgesic overuse showed higher values in each concept of the SF-36 than those with analgesic overuse. Despite the low proportion of abusers in this study, differences were significant for physical functioning and bodily pain. CONCLUSIONS: In the general population, CDH reduced all QoL aspects studied with the SF-36. This reduction in QoL was most marked in subjects with analgesic overuse. QoL was affected more by the chronicity than by the intensity of pain. PMID- 11940694 TI - Transcranial magnetic stimulation of visual area V5 in migraine. AB - OBJECTIVE: To examine visual cortical excitability in persons with migraine using transcranial magnetic stimulation (TMS) over an extrastriate area of the brain, area V5. BACKGROUND: Previous studies found that persons with migraine have a lower phosphene threshold than healthy control subjects with TMS delivered over the primary visual cortical area V1. The result suggests that the occipital cortex in migraineurs between migraine attacks is hyperexcitable. However, it is not known whether interictal cortical hyperexcitability is also present in areas of the association visual cortex. METHOD: To investigate this, single-pulse TMS was delivered over visual area V5, the motion cortex, to 16 persons with migraine and visual aura, nine migraineurs without visual aura, and 16 healthy control subjects. TMS was delivered at intensities ranging from 30 to 100% of maximum stimulator output or until the participant reported seeing phosphenes (visual illusions characterized by flashes of light). Thresholds to phosphenes were obtained for each participant using a staircase procedure. RESULT: Significantly lower phosphene thresholds for TMS delivered over V5 were found in migraineurs as compared with control subjects. Qualitatively, the migraineurs' experience of phosphenes were more vivid, florid, and sustained compared with that of control subjects. CONCLUSION: Results of this study indicate that hyperexcitability of the visual cortex in migraine goes beyond visual area V1 and demonstrates for the first time a significant difference in threshold for excitability of visual area V5 in persons with migraine. PMID- 11940695 TI - Incidence and mortality of generalized convulsive status epilepticus in California. AB - BACKGROUND: Few population-based studies of status epilepticus have been performed in the United States. OBJECTIVE: To determine the incidence, case fatality, and demographics of generalized convulsive status epilepticus (GCSE) in the state of California. METHODS: Using a state-wide hospital discharge database, the authors identified all hospitalizations from 1991 through 1998 with a discharge diagnosis of convulsive status epilepticus. They identified the first admission for each individual to estimate the incidence of GCSE. In-hospital case fatality rates were calculated, and multivariate analysis was performed to determine predictors of death during hospitalization. Secondary diagnoses were analyzed by retrieving all discharge diagnoses accompanying the diagnosis of GCSE. RESULTS: The incidence rate of GCSE was 6.2/100,000 population and fell by 42% between the years 1991 and 1998 from 8.5 to 4.9/100,000. The rate of GCSE was highest among children under the age of 5 (7.5/100,000) and among the elderly (22.3/100,000). Blacks also demonstrated a relatively high incidence of GCSE (13.4/100,000). The case fatality for incident admissions was 10.7%, with increasing age being the only significant predictor in multivariate analysis. Case fatality was highest in patients who also carried a diagnosis of anoxia, CNS infection, or stroke. CONCLUSIONS: The incidence of GCSE requiring hospitalization has fallen over the last decade and is lower than that reported in previous studies. The case fatality is also lower than that reported previously. Further studies are needed to determine the cause of this decline in incidence and mortality of GCSE. PMID- 11940696 TI - Seasonal variation in immune measurements and MRI markers of disease activity in MS. AB - BACKGROUND: The exact mechanisms by which T cells contribute to MS progression are not known. Recently, the results of cross-sectional studies suggested seasonal variation of both interferon (IFN)-gamma production and the number of active MRI lesions in MS. OBJECTIVE: To investigate whether seasonal fluctuations of IFN-gamma and active MRI lesions could be confirmed and whether any correlations could be detected. METHODS: Data were analyzed from a group of 28 MS patients in whom detailed longitudinal monitoring of both immune function and MRI measurements had taken place. RESULTS: Significant seasonal variation was observed in T-cell activation as measured by the ability of T cells to secrete the pro-inflammatory cytokines tumor necrosis factor-alpha and IFN-gamma. Maximum values were found in samples obtained during autumn. Even though clear fluctuations were observed, no significant seasonal variation could be detected in the number of active MRI lesions. Fluctuations of in vitro IFN-gamma secretion correlated weakly with changes in active MRI lesions. CONCLUSIONS: The finding of seasonal variation of immune function in serially MRI-monitored MS patients suggests an environmental role in T-cell activation. PMID- 11940697 TI - A pilot randomized trial of oxandrolone in inclusion body myositis. AB - BACKGROUND: Inclusion body myositis (IBM) remains without effective therapy. As anabolic steroids have myotrophic properties, the authors studied whether a synthetic androgen, oxandrolone, would have efficacy in IBM. METHODS: A double blind, placebo-controlled, crossover design was used. Patients received oxandrolone or placebo for 12 weeks followed by a minimum 2-month washout period, followed by 12 weeks of the alternative treatment. Maximal voluntary isometric contraction testing (MVICT), manual muscle testing (MMT), and functional performance testing were obtained before and after each treatment period, with the whole-body MVICT score as the primary outcome measure. RESULTS: Of 19 patients enrolled, 16 (14 men, 2 women; median age 68.5 years) had complete data for at least the first treatment period, with 13 completing the entire study. Whole-body MVICT improved by a median of 15.5 kg with drug and 4.1 kg with placebo (p = 0.06), whereas MMT demonstrated a median increase of 2.0 Medical Research Council points with drug and 0.9 point with placebo (p = 0.33). Upper extremity MVICT demonstrated a significant treatment effect, with strength increasing a median 6.3 kg with drug vs 2.5 kg with placebo (p = 0.006). Stair climbing also increased a median of 1 step on average with drug versus no change with placebo (p < 0.001). Minimal adverse effects occurred. CONCLUSIONS: Oxandrolone had a borderline significant effect in improving whole-body strength and a significant effect in improving upper-extremity strength as measured by MVICT. Given these findings, further study of this drug, possibly in combination with an immunomodulating agent, is warranted. PMID- 11940698 TI - Resting oxygen consumption and in vivo ADP are increased in myopathy due to complex I deficiency. AB - BACKGROUND: Patients with isolated complex I deficiency (CID) in skeletal muscle mitochondria often present with exercise intolerance as their major clinical symptom. OBJECTIVE: To study the in vivo bioenergetics in patients with complex I deficiency in skeletal muscle mitochondria. METHODS: In vivo bioenergetics were studied in three of these patients by measuring oxygen uptake at rest and during maximal exercise, together with forearm ADP concentrations ([ADP]) at rest. Whole body oxygen consumption at rest (VO(2)) was measured with respiratory calorimetry. Maximal oxygen uptake (VO(2)max) was measured during maximal exercise on a cycle ergometer. Resting [ADP] was estimated from in vivo (31)P MRS measurements of inorganic phosphate, phosphocreatine, and ATP content of forearm muscle. RESULTS: Resting VO(2) was significantly increased in all three patients: 128 +/- 14% (SD) of values in healthy control subjects. VO(2)max in patients was on average 2.8 times their VO(2) at rest and was only 28% of VO(2)max in control subjects. Resting [ADP] in forearm muscle was significantly increased compared with healthy control subjects (patients 26 +/- 2 microM, healthy controls 9 +/- 2 microM). CONCLUSION: In patients with CID, the increased whole-body oxygen consumption rate at rest reflects increased electron transport through the respiratory chain, driven by a decreased phosphorylation potential. The increased electron transport rate may compensate for the decreased efficiency of oxidative phosphorylation (phosphorylation potential). PMID- 11940699 TI - Robert Barany and the controversy surrounding his discovery of the caloric reaction. AB - It was not until the latter half of the 19th century that the vestibular part of the inner ear was recognized to have a function separate from hearing. Robert Barany was one of the first clinicians to assess vestibular function systematically in patients. He developed standard caloric, rotational, and pointing tests and was the first to describe benign paroxysmal positional nystagmus. Barany began his career working in the otology clinic of Adam Politzer at the University of Vienna. He received the Nobel Prize in Medicine in 1916 for his work on the mechanism of the caloric reaction. However, in the aftermath of the First World War and the controversy surrounding his selection for the Nobel Prize, Barany left Vienna for Uppsala, Sweden, where he spent the last 20 years of his life in relative isolation. PMID- 11940700 TI - Internal carotid artery "spring sign". PMID- 11940702 TI - Sequence analysis of the COL5A2 gene in patients with spontaneous cervical artery dissections. AB - The authors searched for mutations in the gene that codes for the alpha2 chain of type V procollagen in 10 patients with spontaneous cervical artery dissections (sCAD). Two patients carried a missense mutation affecting the predicted C propeptide (T1227S; D1429V). A third patient carried two mutations (V509A and P830L) in the same alpha2(V) chain. The T1227S mutation and the V509A/P830L haplotype also were detected among 50 healthy subjects. The D1429V substitution was detected neither in a series of 150 healthy control subjects nor among 50 additional patients with sCAD. PMID- 11940701 TI - Do persons with dementia vote? AB - To examine whether patients with dementia voted in the 2000 US Presidential Election, the authors surveyed 75 caregivers of patients with dementia. A substantial portion of patients with mild to moderate dementia voted on their own at a voting booth. Patients cared for by spouses were more likely to vote than patients cared for by adult children. Further research is needed to understand how persons with dementia and their caregivers decide what activities the person can and cannot continue and how well these decisions correspond to measures of competency. PMID- 11940703 TI - Inability to control anger or aggression after stroke. AB - Using the 10-item Spielberger Trait Anger Scale, the authors interviewed 145 patients with stroke regarding inability to control anger or aggression (ICAA). Poststroke depression and emotional incontinence were also assessed. ICAA was present in 47 patients (32%) and was closely related to motor dysfunction, dysarthria, emotional incontinence, and lesions affecting frontal lenticulocapsular-pontine base areas. ICAA seems to be one of the major behavioral symptoms in patients with stroke. PMID- 11940704 TI - Long-term survival after gene therapy for a recurrent glioblastoma. AB - A patient presenting with a recurrent glioblastoma (GBM) survived 3 years after suicide gene therapy and finally died of a disseminated breast cancer with no indication of tumor recurrence on MRI. Postmortem analysis showed no evidence of recurrence of the GBM, neither near the initial tumor localization nor in any other area of the brain. Such an evolution is unusual in the course of this disease and may suggest in this particular case a cure of the GBM. PMID- 11940705 TI - APOE: a potential marker of disease progression in ALS. AB - Although documented in AD, the role of APOE remains unclear in ALS. APOE phenotype and plasma levels were measured in 403 patients with ALS and were correlated with clinical parameters and survival time. No correlations were observed between the APOE phenotype and these variables. In contrast, APOE plasma levels were correlated with both rate of deterioration and survival time and appeared to be an important risk factor for decreased survival time with a relative risk of 0.647 (95% CI: 0.465 to 0.901; p = 0.01). PMID- 11940706 TI - APOE genotype influences acquisition and recall following traumatic brain injury. AB - APOE has been demonstrated to influence traumatic brain injury (TBI) outcome. The relationship between APOE genotype and memory following TBI was examined in 110 participants in the Defense and Veterans' Head Injury Program. Memory performance was worse in those who had an APOE epsilon 4 allele (n = 30) than those who did not (n = 80), whereas genotype groups did not differ on demographic or injury variables or on measures of executive functioning. These data support a specific role for the APOE protein in memory outcome following TBI, and suggest an APOE isoform-specific effect on neuronal repair processes. PMID- 11940708 TI - Frequent mutations of SCN1A in severe myoclonic epilepsy in infancy. AB - Mutations in the neuronal voltage-gated sodium channel alpha-subunit type I gene (SCN1A) were found responsible for severe myoclonic epilepsy in infancy (SMEI). The authors describe novel mutations of SCN1A in Japanese patients with SMEI. They screened 12 unrelated patients and a pair of monozygotic twins and detected 10 mutations that lead to truncation of the protein. PMID- 11940707 TI - Porphyria presenting with bilateral radial motor neuropathy: evidence of a novel gene mutation. AB - The authors identified a novel mutation of the porphobilinogen deaminase (PBG-D) gene in a patient with acute intermittent porphyria presenting with severe and bilateral axonal radial motor neuropathy. Electrophysiologic studies revealed prominent involvement of distal radial nerves in the setting of mild polyneuropathy. Analysis of the PBG-D gene revealed a single base-pair insertion (887insA) in exon 14. PMID- 11940709 TI - Interleukin-2 in the pathogenesis of perinatal white matter damage. AB - Proinflammatory cytokines were reported to be implicated in the pathogenesis of perinatal white matter lesions. The authors document for the first time the in situ detection of interleukin-2 and interleukin-2 receptor (IL-2R) in these human white matter lesions. These results suggest that interleukin-2, reported to be toxic to oligodendrocytes and myelin, could play a role in the molecular cascade leading to white matter damage in periventricular leukomalacia. PMID- 11940710 TI - Brown-Sequard syndrome after esophageal sclerotherapy and crack cocaine abuse. PMID- 11940711 TI - Triparesis: MRI documentation of bipyramidal medullary infarction. PMID- 11940712 TI - Opsoclonus-myoclonus syndrome following Epstein-Barr virus infection. PMID- 11940713 TI - Transdermal nicotine in PD: a randomized, double-blind, placebo-controlled study. PMID- 11940714 TI - Visual loss in cysticercosis: analysis of 23 patients. PMID- 11940715 TI - Fatigue is not associated with raised inflammatory markers in multiple sclerosis. PMID- 11940716 TI - Prospective study of local cutaneous reactions in patients receiving IV phenytoin. PMID- 11940717 TI - Cholesterol and Alzheimer's disease: is there a link? PMID- 11940718 TI - Prolonged cortical silent period after transcranial magnetic stimulation in generalized epilepsy. PMID- 11940719 TI - Very late onset Friedreich's presenting as spastic tetraparesis without ataxia or neuropathy. PMID- 11940723 TI - A national random survey of bone mineral density reporting in the United States. AB - The rapidly evolving technology of bone mineral density (BMD) testing has revolutionized the clinical care of osteoporosis; however, at present, there are no guidelines for BMD reporting. A survey was mailed to a random sample of bone densitometry centers in the United States registered in the National Osteoporosis Foundation database in order to evaluate the practice of BMD reporting in the United States. Of the 1200 questionnaires mailed, 22.5% were completed and returned. Spine and hip BMD were routinely measured at 71% of the centers and were expressed as T-scores by 90% of centers. The World Health Organization working group definition of osteoporosis was included in the report by 64% of the survey responders and was used as the sole criterion to make treatment recommendations by 34%. Fracture risk was reported by 70% of the centers and only the minority (<15%) applied appropriate age and gender restrictions. There were geographic and specialty variations in the practices of bone density reporting. Despite the established value of clinical densitometry in the care of patients at risk for osteoporosis, our survey revealed that clinical information, including fracture risk, was missing from many reports. A re-examination of the practice of clinical densitometry reporting is warranted. PMID- 11940724 TI - Relationship between phalangeal bone density and risk of vertebral fracture. AB - The aims of our study were to determine the relationship between bone mineral density (BMD) measurements of the phalanges obtained with the accuDEXA and recent vertebral fractures. To determine whether osteoarthritis of the hands affects phalangeal BMD measurements, and to illustrate the conversion of phalangeal BMD measurements to absolute fracture risk estimates for clinical application. The prospective Hawaii Osteoporosis Study began in 1981, and incident vertebral fractures were identified from serial radiographs obtained at approx 2-yr intervals. Vertebral fractures occurring between 1993 and 1994 and 1997 and 1998 were compared to phalangeal BMD measurements obtained in 1997-1998. A total of 199 women participated in this case-control study. The association of the phalangeal BMD measurements with vertebral fractures was examined in age adjusted, logistic regression models. Results are expressed as odds ratios (ORs) per SD difference in the phalangeal BMD measurements. Osteoarthritis of the hands was graded according to the Kellgren-Lawrence scale. There were 34 incident fractures since the eighth examination in 1993-1994. For vertebral fractures, the OR per SD of phalangeal BMD was 1.5 (1.0-2.1). Phalangeal BMD was not influenced significantly by established osteoarthritis (p = 0.68). Phalangeal BMD measurements obtained with the accuDEXA device relate to recent vertebral fractures and can be used to identify women at high risk of fractures. The phalangeal BMD measurements obtained with this device are not significantly influenced by the presence of osteoarthritis of the hands. PMID- 11940725 TI - Lumbar spine bone measurements in infants: whole-body vs lumbar spine dual X-ray absorptiometry scans. AB - Availability of software delineation of the lumbar spine region from infant whole body (IWB) dual energy X-ray absorptiometry scan offers an opportunity to gain additional information at the lumbar spine without a separate scan, although the validity of this technique has never been tested. Lumbar spine measurements derived from IWB scans using software-delineated first to fourth lumbar vertebrae, and from specific infant spine (IS) scans, were determined in 111 infants using two pencil-beam densitometers. Intraoperator repeatability determined by reanalysis of 10 pairs of IWB and IS scans from each densitometer. Lumbar spine area, bone mineral content, and bone mineral density from IWB and IS scans were significantly correlated r = 0.68-0.95, p < 0.001 for all comparisons) but show poor agreement (Bland-Altman method) with one SD of the differences equal to 26-55% of the mean. Intraoperator reanalysis shows good agreement with one SD of the differences from IWB scans at <7% of the mean, and <2.9% from IS scans. Findings were the same for both densitometers. We conclude that lumbar spine bone measurements from IWB or IS scans are highly reproducible and significantly correlated but data from IWB scans cannot substitute for data from IS scans. PMID- 11940726 TI - Effect of two forms of alendronate administration upon bone mass after two years of treatment. AB - The efficacy of alendronate in slowing the loss of bone mass, or even in increasing it, in osteoporotic patients and thus reducing the risk of new fractures has been described. Nevertheless, the way of taking this drug, together with its side effects, sometimes produces withdrawals. In this study, we analyzed if an alternative way of taking the alendronate improves the follow-up of the treatment and if it had the same effect on bone mineral metabolism than the traditional way of prescription. An open, intention-to-treat study, with follow up of 2 yr was conducted. Eighty women suffering from postmenopausal osteoporosis were included in the study. They were classified in a random manner into two groups, each one of them received 10 mg/d alendronate, together with 1.2 g of calcium and 800 IU of Vitamin D3. Group I received the drug fasting, before breakfast, as usually prescribed and group II received the alendronate fasting, at noon, before lunch. Biochemical markers of bone remodeling were determined. Total alkaline phosphatase, osteocalcin, tartrate-resistant acid phosphatase, urine calcium/creatinine ratio, crosslinked N-telopeptides of type I collagen/creatinine ratio, serum calcium, and parathyroid hormone were also determined, and a lateral dorsolumbar radiography of the spine was performed. Bone mineral density was determined in the lumbar spine by dual-energy X-ray absorptiometry and quantitative computed tomography and by dual-energy X-ray absorptiometry in the proximal femur. Both groups showed an increase in bone mineral density in the lumbar spine and in the proximal femur, which was statistically significant after 1 yr of treatment in the range between 1.5% and 4.3%, depending on the anatomical localization where bone mineral density was measured. There was also an important decrease in the biochemical markers of bone remodeling, between 5.6% and 42.5%, depending on the biochemical marker; the decrease of amino-terminal telopetide during the first year was more important. The group that received alendronate in the morning reported a significantly higher number of withdrawals than the group that received the drug at noon. The alternative administration of 10 mg alendronate at noon had the same effect on bone mineral metabolism than its traditional administration in the morning, but the rate of withdrawals was significantly lower. PMID- 11940727 TI - Reproducibility of DXA in obese women. AB - Dual-energy X-ray absorptiometry (DXA) measurements were analyzed using two versions of software (Hologic V8.1a and V8.21) to compare the short- and long term precisions of the measurements. Software V8.21 was designed by the manufacturer to better address magnification effects on estimations of soft tissue lean mass. Twenty weight-stable, obese postmenopausal Caucasian women aged 40-70 yr participated in the study. Total and regional body composition measurements were obtained at baseline and after 3 mo, using a fan beam Hologic QDR 4500A absorptiometer. For the estimation of precision, duplicate scans obtained on the same day for nine women were analyzed using both versions of the software. The correlations between duplicate scans ranged from 0.886 to 0.998 and were similar between software versions. The CVs for fat and lean weights and bone mineral content (BMC) were 1.2%, 1.1%, and 1.7%, respectively, for software V8.21 compared to 1.3%, 1.3%, and 2.1%, respectively, for V8.1a. Systematic differences were found between software versions with higher values for fat and lean weights for software version V8.21. The 3-mo, long-term reproducibility of body composition estimates from DXA was only slightly less than short-term reproducibility for both software versions (coefficient of variation [CV] range from 1.3% for BMC weight to 11.0% for arm fat). Software V8.21 yielded smaller percentage mean differences between scale and DXA-estimated weights (-2.4% and 7.2% at baseline and -2.9% and -7.6% at 3 mo, respectively) and higher fat and lean weights (49.12 and 47.1 kg and 49.6 and 44.6 kg, respectively) than V8.1a. Reproducibility of all variables was comparable between software versions. PMID- 11940728 TI - Determination of bone size of hip, spine, and wrist in human pedigrees by genetic and lifestyle factors. AB - Osteoporosis is a major public health problem defined as a loss of bone strength, of which bone size is an important determinant. Compared with extensive studies on bone mass, studies on the importance of factors determining variation in bone size are relatively few. In particular, the significance of genetic factors is largely unknown. In 49 pedigrees with 703 subjects bone sizes of the hip, spine, and wrist were measured by dual X-ray absorptiometry. We evaluated the contribution of genetic factors in determining variation in bone size of the hip, spine, and wrist while studying age, sex, weight, height, exercise, smoking, alcohol consumption, and the interaction among these factors as covariates for their effects on bone size. We found that, on average, males have larger bone sizes. Male bone sizes at the spine and hip increased with age; however, the effect of age in our female subjects was nonsignificant. Height invariably affected bone size at all the sites studied. Alcohol consumption and exercise generally had significant effects in increasing bone size at the spine and/or hip in both males and females. After adjusting for sex, age, weight, height, lifestyle factors, and the significant interactions among these factors, heritabilities (+/-SE) were, respectively, 0.48 (0.09), 0.64 (0.08), and 0.60 (0.09) for bone size at the hip, spine, and wrist. PMID- 11940729 TI - Bone mineral density assessment: comparison of dual-energy X-ray absorptiometry measurements at the calcaneus, spine, and hip. AB - It is widely accepted that bone mineral density (BMD) measurements obtained by dual-energy X-ray absorptiometry (DXA) at the spine, hip, and calcaneus predict fracture risk. Few published studies to date have examined the relationship between pDXA measurements at the calcaneus to those at the hip and spine. It has been demonstrated that T-score-based criteria cannot be universally applied to all skeletal sites and measurement technologies. Our goal was to define the calcaneal T-score threshold equivalent to low bone mass at the hip or spine. A total of 119 female patients between the ages of 33 and 76 yr of age were recruited at Boston University Medical Center for bone densitometry screening. Bone density measurements were obtained at the calcaneus using the portable Norland Apollo Densitometer (Norland Medical Systems, Fort Atkinson, WI) and at the hip and spine using the Norland Eclipse densitometer. By defining a pDXA T score < or =-1 as a positive test and DXA scores < or =-1 as the presence of low bone mass, we obtained a specificity of 100% and a sensitivity of 73% (positive predictive value 100% and negative predictive value 80%) in detecting low bone mass at the femoral neck in women over age 65 yr. In women between 40 and 65 yr of age, we obtained a sensitivity of 50% and a specificity of 93% (positive predictive value 93% and negative predictive value 50%) in detecting low bone mass at the femoral neck. In women less than 40 yr of age, we obtained a sensitivity of 13% and a specificity of 100% (positive predictive value 100% and negative predictive value 75%) in detecting low bone mass at the femoral neck. From receiver operating characteristic curves, a calcaneal T-score < or =0.0 detects those with a T-score < or =-1 at the femoral neck and lumbar spine with 100% and 85% sensitivity, respectively. Peripheral DXA of the calcaneus is a sensitive and specific test to diagnose low bone mass in women over 65 yr of age. In women under 65 yr of age, this modality, though not as sensitive, is specific in detecting low bone mass. We conclude that a pDXA calcaneal T-score < or =0 is highly sensitive in predicting osteopenia and osteoporosis at the femoral neck and lumbar spine. PMID- 11940730 TI - North American male reference population for speed of sound in bone at multiple skeletal sites. AB - Alternatives to dual-energy X-ray absorptiometry (DXA) have been sought to increase access to low-cost osteoporosis risk assessment. Early quantitative ultrasound (QUS) systems measured speed of sound (SOS) and broadband ultrasound attenuation (BUA) at the calcaneus, and these were demonstrated to be good predictors of hip fracture risk. Recent studies have demonstrated the usefulness of other peripheral sites to assess bone status. The Sunlight Omnisense (Sunlight Medical, Rehovot, Israel) is a portable, inexpensive QUS device capable of multiple-site SOS measurement. To provide a robust male reference database, 588 healthy Caucasian males aged 20-90 yr were recruited from 6 centers across North America. SOS measurements were taken at the distal 1/3 radius, proximal third phalanx, midshaft tibia, and fifth metatarsal. A female reference database has previously been collected at North American sites. The results indicate that SOS in males exhibits an age-related decline beginning in the fifth decade at the radius, phalanx, and metatarsal, whereas the tibial SOS remains nearly constant until the ninth decade. Although females reach a higher-peak SOS than males at most sites, SOS is higher in males at all sites after the sixth decade, as a result of a more gradual decline in SOS. Longitudinal monitoring of healthy men should be performed to confirm these cross-sectional results. PMID- 11940731 TI - Densitometric study of human developing dry bones: a review. AB - In this article we describe the use of dual-energy X-ray absorptiometry (DXA) technique in the study of fetal and infant bone development. In particular, we review the results of densitometric studies on human developing dry bones such as spine, femur, and mandible; these data are compared with the results obtained with other methods used in the past. In particular DXA technique applied in dry is able to detect the first appearances of the ossification centers; show individual variability of bone mineral density yet during the prenatal life; and determine the correlation between bone mineral density and the variation of mechanical loading. PMID- 11940732 TI - Dual X-ray absorptiometry of hip, heel ultrasound, and densitometry of fingers can discriminate male patients with hip fracture from control subjects: a comparison of four different methods. AB - Few studies have examined different bone densitometry techniques to determine male hip fracture risk. We conducted a case-control study of 31 noninstitutionalized men, mean age 77 yr, with a first hip fracture and compared the results with 68 randomly selected age-matched control subjects. The methods used were dual X-ray absorptiometry (DXA) of the proximal femur, quantitative ultrasound (QUS) of the heel and fingers, and radiographic absorptiometry of the fingers. Case patients had significantly lower values (4-17%; p < 0.01) for all methods. The odds ratios for every SD reduction in bone values were 4.8 (95% confidence interval [CI]: 2.3-9.9) for DXA of the femoral neck, 2.2 (95% CI: 1.2 3.9) for QUS of the heel, 2.0 (95% CI: 1.2-3.3) for QUS of the phalanges, and 3.1 (95% CI: 1.5-6.6) for radiographic absorptiometry of the phalanges. The results indicate a strong capability of DXA of the femoral neck to distinguish between men with a first hip fracture and control subjects. Furthermore, ultrasound of the heel and fingers as well as radiographic absorptiometry proved capable of discriminating men with hip fractures from control subjects. PMID- 11940733 TI - Implications in the use of T-scores for the diagnosis of osteoporosis in men. AB - Osteoporosis is recognized as a disorder of both men and women. However, the World Health Organization's (WHO) definition of osteoporosis (a bone mineral density [BMD] T-score of -2.5 or less) was formulated for use with postmenopausal women only. In the absence of a BMD-based definition for male osteoporosis, the WHO definition is often applied to men as well. Several important questions exist when considering the use of T-scores in men. First, is the WHO definition appropriate for men? What is the impact of using a -2.5 criteria, in terms of the number of men that would be identified as osteoporotic? When calculating T-scores in men, should male or female young normal values be used? Can the same T-score criteria be used for all skeletal sites and technologies? To address these questions, osteoporosis prevalence estimates for men aged 50 yr and over were generated using WHO methods and manufacturer normative data from dual-energy X ray absorptiometry (DXA), quantitative computed tomography (QCT), and ultrasound. Estimates were determined for several skeletal sites and technologies using both male and female young normal values. Prevalence estimates were compared to published fracture risk estimates. Mean T-scores declined with age at all measurement sites. Discrepancies were found between the different skeletal sites and techniques, similar to the previously reported differences in women. A -2.5 criterion (based on young normal males or females) appeared to underestimate the prevalence of osteoporosis, except for QCT, which seemed to overestimate risk. Depending on the technique used, 0 to 12.5 million US men 50 yr of age and older would be classified as osteoporotic using the WHO definition. T-Scores based on male norms were less discordant across skeletal sites than female-based T-scores. Male-based T-scores between -1.8 and -2.3 using DXA and ultrasound and -3.1 for QCT provided osteoporosis prevalence estimates that approximated the likelihood of common fractures in men 50 and over. We conclude that the use of single T score-based criterion for the diagnosis of osteoporosis in men has many potential difficulties. BMD measurement techniques provide discrepant estimates of prevalence and may underestimate the size of the male population at risk for fracture. Based on available normative data, a -2.5 criterion underestimates osteoporosis prevalence in men, whether based on male or female norms. Prospective studies are needed to further refinement to the BMD definition of osteoporosis in men. PMID- 11940734 TI - Discrepancy in results between spine and hip scans of a woman with end stage renal disease. AB - Conditions and artifacts such as aortic calcifications, osteophytes, hip prostheses, and metallic objects can affect the results of dual X-ray absorptiometry scans of the spine and hip. Abdominal surgery often entails the use of metal sutures causing subtle artifacts near or over the lumbar spine resulting in inaccurate bone mineral density (BMD) measurements. We herein report a case of a woman whose spine BMD appeared normal while her hip BMD was > -3.5 SDs. Although the abdominal artifacts create some uncertainty in the diagnosis, renal osteodystrophy is suspect owing to the patient's renal history. This case demonstrates the importance of acquiring scans at two or more sites, closely evaluating scans for artifacts, and obtaining the patient's medical history. PMID- 11940736 TI - Invisible excellence: the Presidential Address from the 31st Congress of the Society of Critical Care Medicine. PMID- 11940737 TI - Continuous monitoring of cerebrovascular pressure reactivity allows determination of optimal cerebral perfusion pressure in patients with traumatic brain injury. AB - OBJECTIVES: To define optimal cerebral perfusion pressure (CPPOPT) in individual head-injured patients using continuous monitoring of cerebrovascular pressure reactivity. To test the hypothesis that patients with poor outcome were managed at a cerebral perfusion pressure (CPP) differing more from their CPPOPT than were patients with good outcome. DESIGN: Retrospective analysis of prospectively collected data. SETTING: Neurosciences critical care unit of a university hospital. PATIENTS: A total of 114 head-injured patients admitted between January 1997 and August 2000 with continuous monitoring of mean arterial blood pressure (MAP) and intracranial pressure (ICP). MEASUREMENTS AND MAIN RESULTS: MAP, ICP, and CPP were continuously recorded and a pressure reactivity index (PRx) was calculated online. PRx is the moving correlation coefficient recorded over 4-min periods between averaged values (6-sec periods) of MAP and ICP representing cerebrovascular pressure reactivity. When cerebrovascular reactivity is intact, PRx has negative or zero values, otherwise PRx is positive. Outcome was assessed at 6 months using the Glasgow Outcome Scale. A total of 13,633 hrs of data were recorded. CPPOPT was defined as the CPP where PRx reaches its minimum value when plotted against CPP. Identification of CPPOPT was possible in 68 patients (60%). In 22 patients (27%), CPPOPT was not found because it presumably lay outside the studied range of CPP. Patients' outcome correlated with the difference between CPP and CPPOPT for patients who were managed on average below CPPOPT (r =.53, p <.001) and for patients whose mean CPP was above CPPOPT (r = -.40, p <.05). CONCLUSIONS: CPPOPT could be identified in a majority of patients. Patients with a mean CPP close to CPPOPT were more likely to have a favorable outcome than those whose mean CPP was more different from CPPOPT. We propose use of the criterion of minimal achievable PRx to guide future trials of CPP oriented treatment in head injured patients. PMID- 11940738 TI - Fluid thresholds and outcome from severe brain injury. AB - OBJECTIVE: To determine, by retrospective analysis, critical thresholds for intracranial pressure, mean arterial pressure, cerebral perfusion pressure, and fluid balance associated with poor outcome in patients with severe brain injury. DESIGN: Retrospective review of patient data from the prospective, randomized, multicenter National Acute Brain Injury Study: Hypothermia, comparing outcome results at 6 months after injury with intracranial pressure, mean arterial pressure, cerebral perfusion pressure, and fluid balance measurements recorded during the 96-hr period after randomization. SETTING: Emergency departments and intensive care units in 11 metropolitan tertiary care university hospitals. PATIENTS: A total of 392 patients, aged 16-65 yrs, with severe, nonpenetrating brain injuries and a Glasgow Coma Scale score of 3-8 after resuscitation, who were enrolled in a study designed to determine the treatment effect of moderate hypothermia in patients with severe brain injury. INTERVENTION: Standard brain injury treatment for 193 randomly assigned patients and standard treatment plus hypothermia for 48 hrs for 199 patients. MEASUREMENTS AND MAIN RESULTS: Intracranial pressure levels of 20, 25, and 30 mm Hg, mean arterial pressure levels of 70 and 80 mm Hg, cerebral perfusion pressure levels of 50, 60, and 70 mm Hg, and fluid balance levels in quartiles were examined for their effect on outcome as measured by the Glasgow Outcome Scale at 6 months after injury. When considered separately, any of the following-intracranial pressure >25 mm Hg, mean arterial pressure <70 mm Hg, or cerebral perfusion pressure <60 mm Hg and fluid balance lower than -594 mL-was associated with an increased percentage of patients with poor outcome. When the variables were combined into a stepwise logistic regression model, Glasgow Coma Scale score at admission, age, mean arterial pressure <70 mm Hg, fluid balance lower than -594 mL, and intracranial pressure > 25 mm Hg, in that order, were the most powerful variables in determining outcome. CONCLUSIONS: Exceeding thresholds of intracranial pressure, mean arterial pressure, cerebral perfusion pressure, and fluid volume may be detrimental to severe brain injury outcome. Fluid balance lower than -594 mL was associated with an adverse effect on outcome, independent of its relationship to intracranial pressure, mean arterial pressure, or cerebral perfusion pressure. PMID- 11940739 TI - Patients' recollections of stressful experiences while receiving prolonged mechanical ventilation in an intensive care unit. AB - OBJECTIVE: To describe stressful experiences of adult patients who received mechanical ventilation for > or =48 hrs in an intensive care unit. DESIGN: Prospective cohort study. SETTING: Four intensive care units within an East Coast tertiary-care university medical center. PATIENTS: Patients were 150 adult intensive care unit patients receiving mechanical ventilation for > or =48 hrs. INTERVENTION: None. MEASUREMENTS AND MAIN RESULTS: As part of a study of the long term outcomes of adult patients requiring prolonged mechanical ventilation, we used a 32-item questionnaire to collect data on patients' stressful experiences, both psychological (e.g., fearfulness, anxiety) and physical (e.g., pain, difficulty breathing), associated with the mechanical ventilation endotracheal tube and with being in an intensive care unit. Of 554 patients who met study criteria and survived prolonged mechanical ventilation, 150 consented and were oriented to person, place, and situation. Two thirds of these patients remembered the endotracheal tube and/or being in an intensive care unit. The median numbers of endotracheal tube and intensive care unit experiences remembered were 3 (of 7) and 9 (of 22), respectively. If a patient remembered an experience in the questionnaire, it was likely to be moderately to extremely bothersome. Some of the items that many patients found to be moderately to extremely bothersome were pain, fear, anxiety, lack of sleep, feeling tense, inability to speak/communicate, lack of control, nightmares, and loneliness. Stressful experiences associated with the endotracheal tube were strongly associated with subjects' experiencing spells of terror, feeling nervous when left alone, and poor sleeping patterns. CONCLUSIONS: Subjects were more likely to remember experiences that were moderately to extremely bothersome. This might be because the more bothersome experiences were easier to recall or because most of these experiences are common and significant stressors to many of these patients. In either case, these data indicate that these patients are subject to numerous stressful experiences, which many find quite bothersome. This suggests the potential for improved symptom management, which could contribute to a less stressful intensive care unit stay and improved patient outcomes. PMID- 11940740 TI - Time course of platelet counts in critically ill patients. AB - BACKGROUND: Although thrombocytopenia in the intensive care unit (ICU) is associated with a poorer outcome, the precise relationship between the time course of platelet counts and the mortality rate has not been well defined. OBJECTIVE: To describe the time course of the platelet count in relation to the mortality rate in critically ill patients. DESIGN: Substudy of a prospective, multicenter, observational cohort analysis. SETTING: Forty ICUs in 16 countries from Europe, America, and Australia. PATIENTS: Data were collected from all ICU admissions in a 1-month period, excluding patients younger than 12 yrs old and those who stayed in the ICU for <48 hrs after uncomplicated surgery. A total of 1,449 critically ill patients were enrolled, including 257 who stayed in the ICU for >2 wks. INTERVENTIONS: None. MEASUREMENTS: Platelet counts were collected daily throughout the ICU stay, together with other measures of organ dysfunction. Thrombocytopenia was defined as a platelet count of <150 x 103/mm3. A relative increase in platelet count was defined as a 25% increase above the admission value, together with an absolute platelet count of > or =150 x 103/mm3. MAIN RESULTS: For the entire population, the platelet count was lower in the 313 nonsurvivors than in the 1,131 survivors throughout the ICU course. Of the 257 patients who stayed in the ICU for >2 wks, 187 (64%) survived. The platelet count decreased significantly in the first days after admission to reach a nadir on day 4 in both survivors and nonsurvivors. In the survivors, the platelet count returned to the admission value by the end of the first week and continued to rise to become significantly greater than the admission value by day 9. In the nonsurvivors, the platelet count also returned to the admission value after 1 wk, but there was no subsequent increase in platelet count. A total of 138 (54%) patients had thrombocytopenia on day 4, and these patients had a greater mortality rate than the other patients (33% vs. 16%; p <.05). On day 14, 51 (20%) patients had thrombocytopenia, and these patients had a greater mortality rate than the other patients (66% vs. 16%; p <.05). Thrombocytopenia was less common on day 14 than on day 4 (20% vs. 54%; p <.05), but the mortality rate was greater in the thrombocytopenic patients on day 14 than those who were thrombocytopenic on day 4 (66% vs. 33%; p <.05). The ICU mortality rate of nonthrombocytopenic patients on day 14 was also significantly lower in patients with, than without, a relative increase in platelet count on day 14 (11% vs. 30%; p <.05). CONCLUSION: Platelet count changes in the critically ill have a biphasic pattern that is different in survivors and nonsurvivors. Late thrombocytopenia is more predictive of death than early thrombocytopenia. A relative increase in platelet count after thrombocytopenia was present in survivors but not in nonsurvivors. Although a single measured platelet count is of little value for predicting outcome, changes in platelet count over time are related to patient outcome. PMID- 11940741 TI - Plasma procalcitonin and C-reactive protein in acute septic shock: clinical and biological correlates. AB - OBJECTIVE: To determine the relationship between plasma procalcitonin (PCT) levels, C-reactive protein (CRP), white blood cell count (WBC), ionized calcium (Ca2+), and patient outcome; and to compare the diagnostic and prognostic information provided by PCT and by CRP. DESIGN: Prospective, observational study. SETTING: Intensive care unit. PATIENTS: Fifty-three patients with septic shock, consecutively diagnosed according to consensus guidelines. INTERVENTIONS: None. MEASUREMENTS AND MAIN RESULTS: Blood was sampled at diagnosis and 24 and 48 hrs later and in a subgroup (n = 23) after 120 hrs. PCT was measured with LUMItest and CRP with Vitros slides. Ca2+ was calculated according to McLean-Hastings from calcium and protein levels on Vitros. In all 53 patients, PCT and CRP were elevated (>0.5 ng/mL and >10 mg/L, respectively) within 24 hrs after diagnosis. Nonsurvivors (n = 25) were older (p <.001) and had higher Acute Physiology and Chronic Health Evaluation (APACHE) II scores (p =.02) at diagnosis but did not differ in sepsis etiology, medical history, sex ratio, levels of PCT, CRP, and Ca2+, or WBC count at any time point. Using logistic regression, initial PCT levels were correlated with CRP values (p =.001) and APACHE II score (p <.05), but not with age, gender, Ca2+ levels, survival, or type of pathogen. Within 48 hrs, however, PCT levels decreased more frequently from baseline in survivors than in nonsurvivors (80% vs. 41%, p <.05). Likewise, CRP levels decreased more often in survivors (100% vs. 64%, p <.05) but only at 120 hrs. CONCLUSIONS: PCT levels were correlated with the severity of disease at onset (APACHE II) and inflammation (CRP) but not with Ca2+ levels. Inaugural PCT or CRP levels per se poorly predicted outcome but decreasing levels were associated with a higher probability of survival. In this respect, PCT was found to be an earlier marker than CRP. PMID- 11940742 TI - Frequent development of lupus anticoagulants in critically ill patients treated under intensive care conditions. AB - OBJECTIVE: To investigate how often a prolongation of the activated partial thromboplastin time in critically ill patients is caused by lupus anticoagulants and to identify possible triggering events. DESIGN: Prospective study. SETTING: Internal medicine intensive care unit (University Hospital of Vienna, Vienna, Austria). PATIENTS: Fifty-one critically ill patients without severe coagulopathy, hepatopathy, or anticoagulant treatment (35 male, 16 female, median age 60 yrs, range: 22-85 yrs). INTERVENTIONS: All patients were screened daily for lupus anticoagulants with the activated partial thromboplastin time STA assay. MEASUREMENTS AND MAIN RESULTS: Diluted Russell's viper venom time, plasma mixing studies, and confirmation assays were used to identify lupus anticoagulants at the time of an unexplained prolongation of the activated partial thromboplastin time. The influence of heparin was excluded by determination of thrombin clotting time and anti-Xa activity. In 27 of 51 patients (52.9 %) lupus anticoagulants were found after a median stay of 13 days. None of the patients had concomitant immune thrombocytopenia, hypoprothrombinemia, bleeding, or thromboembolic complications. Sepsis (p =.006) and/or catecholamine treatment (p =.002) were significantly associated with the development of lupus anticoagulants. Extracorporeal circulation, transfusion of blood products, or surgery did not increase this risk. Lupus anticoagulants resolved spontaneously in 63% of the patients after a median stay of 17 days. CONCLUSIONS: Lupus anticoagulants are frequent in critically ill patients and associated with sepsis syndrome and/or catecholamine treatment. The prolonged activated partial thromboplastin time does not warrant the administration of coagulation factors or the cessation of anticoagulant therapy or prophylaxis, inasmuch as this phenomenon is not associated with bleeding or thromboembolic complications. PMID- 11940743 TI - Deep vein thrombosis during prolonged mechanical ventilation despite prophylaxis. AB - OBJECTIVE: To determine the prevalence of deep vein thrombosis (DVT) among patients requiring prolonged mechanical ventilation in the intensive care unit. DESIGN: Prospective cohort study. SETTING: Medical intensive care unit of a university-affiliated urban teaching hospital. PATIENTS: Patients requiring mechanical ventilation for >7 days. INTERVENTIONS: All patients admitted to the medical intensive care unit requiring prolonged mechanical ventilation underwent duplex ultrasonography of their lower extremities and upper extremities every 7 days. The main outcome identified was the presence of DVT. Secondary outcomes included hospital mortality, hospital and intensive care unit lengths of stay, and the occurrence of pulmonary embolism. MEASUREMENTS AND MAIN RESULTS: A total of 110 patients requiring mechanical ventilation for >7 days were enrolled. Prophylaxis against DVT was employed in 110 of the patients (100%). A total of 26 patients (23.6%) developed DVT. Patients with DVT were statistically more likely to have underlying malignancy (30.8% vs. 8.3%; p =.004) and longer durations of central venous catheterization (26.9 +/- 22.2 days vs. 14.5 +/- 12.1 days; p =.024) compared with patients without DVT. There were no statistically significant differences in hospital mortality or lengths of stay in the hospital and intensive care unit for patients with and without DVT. Patients documented to have DVT by using duplex ultrasonography had a statistically greater frequency of subsequent pulmonary embolism during their hospitalization (11.5% vs. 0.0%; p =.012). CONCLUSION: The occurrence of DVT is common among patients requiring prolonged mechanical ventilation in the intensive care unit setting despite the use of prophylaxis measures. These data suggest that alternative strategies for the prevention of DVT should be evaluated. Additionally, early detection methods should be considered to reduce the potential morbidity associated with untreated DVT in this high-risk population. PMID- 11940744 TI - Three patients who asked that life support be withheld or withdrawn in the surgical intensive care unit. AB - OBJECTIVE: To illustrate some of the difficulties encountered in the transition from restorative to palliative care and in the pursuit of patient autonomy in the intensive care unit. DESIGN: A review of the cases of three patients who asked that life support be withheld or withdrawn, and a discussion of the legal and ethical implications of these cases. SETTING: A surgical intensive care unit in an academically affiliated public hospital serving as a Level I trauma center. PATIENTS: Three adults with severe injuries who lacked legal surrogates and lost the ability to make medical decisions. INTERVENTIONS: Care from the attending surgery and consultative critical care services, in addition to consultations from the psychiatry service and the hospital ethics committee. MAIN RESULTS: All three patients died after life support was withheld or withdrawn, but not without conflicts among the various services. In two cases, decisions that the patients may or may not have agreed with were made for them by their physicians. In the third case, the patient was not consulted after his clinical situation changed. CONCLUSIONS: Patients' wishes change and are difficult to know, and even when they are known, physicians may have difficulty honoring them. Nevertheless, patient autonomy should be respected whenever possible in the intensive care unit. PMID- 11940745 TI - Deterioration of previous acute lung injury during neutropenia recovery. AB - DESIGN: Although neutropenia recovery is associated with a high risk of deterioration of respiratory condition, no studies designed to identify risk factors for acute respiratory distress syndrome (ARDS) in this situation have been published. SETTING: Medical ICU in a French teaching hospital. SUBJECTS: We conducted a study to describe critically ill cancer patients with ARDS during neutropenia recovery (defined as the 7-day period centered on the day the neutrophil count rose above 1000/mm3 [day 0]) and to compare them with critically ill cancer patients without ARDS during neutropenia recovery. INTERVENTIONS: None. MEASUREMENTS AND MAIN RESULTS: During a 10-yr period, 62 critically ill cancer patients recovered from neutropenia, of whom 21 experienced ARDS during neutropenia recovery, with a median time of -1 days (-2.5-1) between day 0 and ARDS. In-ICU mortality in these 21 patients was 61.9%. As compared with non-ARDS patients, ARDS patients were less likely to have myeloma and more likely to have leukemia/lymphoma treated with adriamycin, a history of pneumonia before neutropenia, and a neutropenia duration >10 days; they had a shorter time since malignancy diagnosis and a longer time from chemotherapy to neutropenia. Neither the leukocyte counts on day 0 nor those during the 6-day neutropenia recovery period were predictive of ARDS. CONCLUSIONS: Patients with acute respiratory failure after prolonged neutropenia complicated by pneumonia are at increased risk for ARDS. PMID- 11940746 TI - Very early extubation in children after cardiac surgery. AB - OBJECTIVE: Very early extubation of children after cardiac surgery has been suggested as a safe alternative to prolonged postoperative intubation but is still not common practice. Studies of early extubation in children may not have described reasons for failure to extubate, or have included nonbypass or only low risk repairs. We present our experience with very early extubation in an inclusive group of children after cardiac surgery. DESIGN: Retrospective chart review. SETTING: University hospital operating room and pediatric intensive care unit (ICU). PATIENTS: A total of 102 consecutive children (age <18 yrs) undergoing cardiac surgery requiring cardiopulmonary bypass. MAIN RESULTS: Forty eight patients were extubated early (88% in the operating room, 12% on arrival in ICU). Patients extubated late were younger (13.8 +/- 26.2 vs. 47.6 +/- 44.5 months), smaller (8.1 +/- 10.7 vs.17.5 +/- 14.2 kg), and had higher ASA scores than patients extubated early (p <.001 for all). The youngest patient extubated early was 2 months old (range, 2-192 months). Paco2 on ICU arrival was higher in the early extubation group (52.4 +/- 6.9 vs. 41.2 +/- 14.7 mm Hg [7.0 +/- 0.9 vs. 5.5 +/- 2.0 kPa], p <.001), and pH was lower (7.27 +/- 0.04 vs. 7.37 +/- 0.16, p <.001). Use of subarachnoid morphine did not affect ability to extubate early. No patients in the early extubation group required special airway support, reintubation, or increased inotropic support after ICU admission. CONCLUSIONS: Successful early extubation of even young children is possible and easily accomplished in most children undergoing cardiopulmonary bypass, even with complex procedures, but advantages of extubation in the operating room vs. immediate ICU extubation remain unclear. Transient mild-to-moderate mixed acidosis is common and requires no treatment. Full implementation requires acceptance by surgical and ICU staffs. PMID- 11940747 TI - Hyponatremia in a pediatric stroke patient: syndrome of inappropriate antidiuretic hormone secretion or cerebral salt wasting? AB - OBJECTIVE: To determine the potential role of atrial natriuretic peptide (ANP) and brain natriuretic peptide (BNP) in the pathogenesis of cerebral salt wasting. DESIGN: Clinical case report. SETTING: Regional pediatric intensive care unit. PATIENT: A 3-yr-old boy with a cerebral infarct secondary to traumatic carotid artery dissection who developed hyponatremia associated with weight loss and excessive renal sodium excretion on the sixth day after hospitalization. MEASUREMENTS AND MAIN RESULTS: Plasma concentrations of ANP, BNP, antidiuretic hormone, and renin were determined serially and compared with concentrations measured in a group of eight healthy children undergoing elective surgical procedures. Compared with controls, ANP and BNP plasma concentrations on the eighth day after hospitalization were increased 1.9-fold and 7.7-fold, respectively. Thereafter, the course of ANP and BNP paralleled that of sodium and H2O excretion and remained elevated until the 14th (BNP) and 16th (ANP) days after hospitalization. Serum antidiuretic hormone and renin concentrations were within normal ranges during the entire observation period. CONCLUSION: Cerebral salt wasting is associated with elevated plasma concentrations of ANP and BNP. Natriuretic peptides may play a role in the pathogenesis of this syndrome. PMID- 11940748 TI - Multicenter, prospective, randomized, single-blind study comparing the efficacy and gastrointestinal complications of early jejunal feeding with early gastric feeding in critically ill patients. AB - OBJECTIVE: To compare the incidence of enteral nutrition-related gastrointestinal complications, the efficacy of diet administration, and the incidence of nosocomial pneumonia in patients fed in the stomach or in the jejunum. DESIGN: Prospective, randomized multicenter study. SETTING: Intensive care units (ICUs) in 11 teaching hospitals. PATIENTS: Critically ill patients who could receive early enteral nutrition more than 5 days. INTERVENTIONS: Enteral nutrition was started in the first 36 hrs after admission. One group was fed with a nasogastric tube (GEN group) and the other in the jejunum through a dual-lumen nasogastrojejunal tube (JEN group). MEASUREMENTS AND MAIN RESULTS: Gastrointestinal complications were previously defined. The efficacy of diet administration was calculated using the volume ratio (expressed as the ratio between administered and prescribed volumes). Nosocomial pneumonia was defined according the Centers for Disease Control and Prevention's definitions. One hundred ten patients were included (GEN: 51, JEN: 50). Both groups were comparable in age, gender, Acute Physiology and Chronic Health Evaluation II, and Multiple Organ Dysfunction Score. There were no differences in feeding duration, ICU length of stay, or mortality (43% vs. 38%). The JEN group had lesser gastrointestinal complications (57% vs. 24%, p <.001), mainly because of a lesser incidence of increased gastric residuals (49% vs. 2%, p <.001). Volume ratio was similar in both groups. A post hoc analysis showed that the JEN group had a higher volume ratio at day 7 than the GEN group (68% vs. 82%, p <.03) in patients from ICUs with previous experience in jejunal feeding. Both groups had a similar incidence of nosocomial pneumonia (40% vs. 32%). CONCLUSIONS: Gastrointestinal complications are less frequent in ICU patients fed in the jejunum. Nevertheless, it seems to be a necessary learning curve to achieve better results with a postpyloric access. Early enteral nutrition using a nasojejunal route seems not to be an efficacious measure to decrease nosocomial pneumonia in critically ill patients. PMID- 11940749 TI - Patient-ventilator interactions during partial ventilatory support: a preliminary study comparing the effects of adaptive support ventilation with synchronized intermittent mandatory ventilation plus inspiratory pressure support. AB - OBJECTIVE: To compare the effects of adaptive support ventilation (ASV) and synchronized intermittent mandatory ventilation plus pressure support (SIMV-PS) on patient-ventilator interactions in patients undergoing partial ventilatory support. DESIGN: Prospective, crossover interventional study. SETTING: Medical intensive care unit, university tertiary care center. PATIENTS: Ten patients, intubated and mechanically ventilated for acute respiratory failure of diverse causes, in the early weaning period, ventilated with SIMV-PS and clinically detectable sternocleidomastoid activity suggesting increased inspiratory load and patient-ventilator dyssynchrony. INTERVENTIONS: Measurement of respiratory mechanics, P0.1, sternocleidomastoid electromyographic activity, arterial blood gases, and systemic hemodynamics in three conditions: 1) after 45 mins with SIMV PS (SIMV-PS 1); 2) after 45 mins with ASV, set to deliver the same minute ventilation as during SIMV-PS; 3) 45 mins after return to SIMV-PS (SIMV-PS 2), with settings identical to those of the first SIMV-PS period. MAIN RESULTS: The same minute ventilation was observed during ASV (11.4 +/- 3.1 l/min [mean +/- sd]) as during SIMV-PS 1 (11.6 +/- 3.5 L/min) and SIMV-PS 2 (10.8 +/- 3.4 L/min). No parameter was significantly different between SIMV-PS 1 and 2, hence subsequent results refer to ASV vs. SIMV-PS 1. During ASV, tidal volume increased (538 +/- 91 vs. 671 +/- 100 mL, p <.05) and total respiratory rate decreased (22 +/- 7 vs. 17 +/- 3 breaths/min, p <.05) vs. SIMV-PS. However, spontaneous respiratory rate increased in six patients, decreased in four, and remained unchanged in one. P0.1 decreased during ASV in all patients except three in whom no change was noted (1.8 +/- 0.9 vs. 1.1 +/- 1 cm H2O, p <.05). During ASV, sternocleidomastoid electromyogram activity was markedly reduced (electromyogram index, where SIMV-PS 1 = 100, ASV 34 +/- 41, SIMV-PS 2 89 +/- 36, p <.02) as was palpable muscle activity. No changes were noted in arterial blood gases, pH, or mean systemic pressure during the trial. CONCLUSION: In patients undergoing partial ventilatory support, with clinical and electromyographic signs of increased respiratory muscle loading, ASV provided levels of minute ventilation comparable to those of SIMV-PS. However, with ASV, central respiratory drive and sternocleidomastoid activity were markedly reduced, suggesting decreased inspiratory load and improved patient-ventilator interactions. These preliminary results warrant further testing of ASV for partial ventilatory support. PMID- 11940750 TI - Systemic endothelial activation is greater in septic than in traumatic hemorrhagic shock but does not correlate with endothelial activation in skin biopsies. AB - OBJECTIVE: Sepsis and severe trauma result in endothelial activation and damage. The activated endothelium expresses adhesion receptors that control leukocyte trafficking. After activation, some adhesion molecules are also released into plasma as soluble forms. The present study was designed to compare the expression of soluble cell adhesion molecules (sCAMs) in three groups of patients: those with septic shock, severe sepsis, and traumatic-hemorrhagic shock. In addition, the endothelial expression of these adhesive molecules was examined in skin biopsies. DESIGN: Prospective observational study SETTING: Intensive care unit at a university hospital PATIENTS: The study included 15 patients with septic shock (by Bone's definition), 11 patients with severe sepsis (by Bone's definition), and 13 patients with traumatic-hemorrhagic shock. Fifteen healthy blood donors served as controls. MEASUREMENTS AND MAIN RESULTS: Measurements of sCAMs were performed on days 1, 2, and 3 of the disease. On day 1, when compared with controls, sE-selectin, sP-selectin, soluble vascular cell adhesion molecule (sVCAM)-1, and soluble intercellular adhesion molecule (sICAM)-1 were markedly elevated in septic shock patients, whereas these sCAMs, except for sP-selectin, were within normal ranges in traumatic-hemorrhagic shock patients. In patients with severe sepsis, an earlier stage than septic shock in the sepsis continuum, intermediate values of sCAMs were found. In skin biopsies of septic shock patients, the endothelial cells expressed a bright staining of constitutive endothelial molecules (CD146, CD144, CD131). Inducible molecules (ICAM-1, VCAM-1, and E-selectin) were positively expressed with bright staining. The biopsies from traumatic-hemorrhagic shock patients showed a similar positive expression of endothelial molecules. CONCLUSION: The patterns of sCAMs indicate that the systemic activation of the endothelium is different in the three clinical entities, maximum in septic shock, intermediate in severe sepsis, and not different from controls in traumatic-hemorrhagic shock. Comparable endothelial activation as evidenced by skin biopsies suggests that caution is required in the interpretation of CAMs in plasma, which does not necessarily reflect the in situ activation state of endothelium. PMID- 11940751 TI - Percutaneous tracheostomy: prospective comparison of the translaryngeal technique versus the forceps-dilational technique in 100 critically ill adults. AB - OBJECTIVE: To compare two different techniques of percutaneous tracheostomy: Griggs' forceps-dilational technique and Fantoni's translaryngeal technique, both performed with the manufacturer's basic kit and with bronchoscopic guidance. DESIGN: A prospective, randomized trial was designed to compare the two tracheostomy techniques. Critically ill patients requiring elective tracheostomy for long-term ventilation were randomized for translaryngeal tracheostomy or forceps-dilational tracheostomy. SETTING: Intensive care unit of a military teaching hospital. PATIENTS: A total of 100 adult patients in the intensive care unit who were mechanically ventilated. PROCEDURES: All tracheostomy procedures were performed at the bedside by using a commercially available set. The procedures were performed by two surgeons, one for bronchoscopic guidance and management of the airway and one for the tracheostomy. MEASUREMENTS AND MAIN RESULTS: The measurements were divided into procedure-related variables (duration, technical difficulties, oxygenation): major and minor complications. The procedure was longer in the translaryngeal technique group (12.9 vs. 6.9 mins, p =.0018). Technical difficulties occurred in 11 patients in the translaryngeal technique group. Uneventful forceps dilational tracheostomy was performed instead. There has been no mortality associated with either technique. Serious complications occurred in one patient in the forceps-dilational technique group (one posterior tracheal wall injury) and in four patients in the translaryngeal technique group (one with a posterior tracheal wall injury and three with severe hypoxia). Significant hypercarbia and acidosis occurred in both the translaryngeal technique group and the forceps-dilational technique group. A significant decrease in Pao2 was observed in the translaryngeal technique group (311 to 261, p =.0069). No bleeding requiring intervention occurred. CONCLUSIONS: Serious complications related to percutaneous tracheostomy occurred in 8.5% and 1.8% of the cases in the translaryngeal technique and the forceps-dilational technique group, respectively (p <.001). Technical difficulties were not rare when using the translaryngeal technique (23%). On the basis of our results, we concluded that the forceps-dilation technique is superior to the translaryngeal technique, with fewer technical difficulties and fewer complications for critically ill patients. PMID- 11940752 TI - Intravenous colforsin daropate, a water-soluble forskolin derivative, prevents thiamylal-fentanyl-induced bronchoconstriction in humans. AB - OBJECTIVE: Forskolin, a direct activator of adenylate cyclase, can relax airway smooth muscle, similar to other agents that increase intracellular cyclic adenine monophosphate. However, the potential usefulness of forskolin in treating bronchospasm is limited by its poor water solubility. Colforsin daropate is a novel and potent water-soluble forskolin derivative. No clinical data have been published on the bronchorelaxant effects of this drug. The aim of this study was to investigate whether intravenous colforsin daropate prevents thiamylal-fentanyl induced bronchoconstriction. DESIGN: Double-blind, prospective, placebo controlled randomized study. SETTING: University teaching hospital. PATIENTS: Thirty-six patients were allocated randomly to two groups: the control group (n = 18) and colforsin daropate group (n = 18). INTERVENTIONS: Intravenous administration of colforsin daropate or placebo (normal saline). MEASUREMENTS AND MAIN RESULTS: Anesthesia was induced with thiamylal 5 mg/kg and vecuronium 0.3 mg/kg. A 15 mg x kg(-1) x hr(-1) continuous infusion of thiamylal followed anesthetic induction. Controlled ventilation was maintained, delivering 50% nitrous oxide in oxygen. Twenty minutes after the induction of anesthesia, the control group patients started to receive 7.5 mL/hr continuous infusion of normal saline, and the colforsin daropate group patients started to receive 0.75 microg x kg(-1) x min(-1) (7.5 mL/hr) continuous infusion of colforsin daropate for 60 min. After that, both groups received fentanyl 5 microg/kg. Systolic and diastolic arterial pressure, heart rate, mean airway resistance (Rawm), expiratory airway resistance (Rawe), and dynamic lung compliance (Cdyn) were measured at the baseline, just before the administration of fentanyl (T30), at three consecutive 6-min intervals after fentanyl injection (T36, T42, and T48) and 30 min after fentanyl injection (T60). At baseline, both groups had comparable Rawm, Rawe, and Cdyn values. In the control group, Rawm increased significantly at T36-60 compared with the baseline, Rawe increased significantly at T36-48 compared with the baseline, and Cdyn decreased significantly at T36-60 compared with the baseline. In the colforsin daropate group, there were no changes in Rawm, Rawe or Cdyn at T36-60. CONCLUSIONS: These observations suggest that intravenous colforsin daropate has a bronchodilator effect in humans. PMID- 11940753 TI - Myocardial inflammatory activation in children with congenital heart disease. AB - OBJECTIVE: In several cardiac-related diseases, there is a strong association between systemic endotoxemia, myocardial cytokine production, and cardiac failure. Because pre- and postoperative endotoxemia recently was reported in children with congenital heart disease, we sought direct evidence of myocardial inflammatory activation in a cohort of children undergoing congenital heart surgery on cardiopulmonary bypass. Inflammatory activation was prospectively defined as the presence of nuclear factor-kappaB nuclear translocation in myocardial tissue samples. DESIGN: Prospective observational study. SETTING: Tertiary care pediatric intensive care unit. PATIENTS: Fifteen children with congenital heart disease undergoing operative repair on cardiopulmonary bypass. INTERVENTIONS: All patients underwent operative repair of congenital heart disease on cardiopulmonary bypass and had plasma samples obtained for endotoxin and tumor necrosis factor-alpha, both pre- and postoperatively. Myocardial tissue samples were obtained intraoperatively, both before and during cardiopulmonary bypass. MEASUREMENTS AND MAIN RESULTS: Elevated plasma endotoxin concentrations were documented in all 15 patients during the study period. In 12 patients, plasma endotoxin was elevated before cardiopulmonary bypass. The median preoperative tumor necrosis factor-alpha concentration was 16.4 pg/mL, which is higher than concentrations reported in adults with New York Heart Association class III congestive heart failure. Examination of myocardial tissue samples revealed nuclear factor-kappaB nuclear translocation (predominantly p50/p65 heterodimers) in nine of 15 patients (60%). Four of these nine patients had nuclear factor-kappaB nuclear translocation before initiation of cardiopulmonary bypass, with p50/p50 homodimers present in two of the four. CONCLUSIONS: These data provide the first evidence of nuclear factor-kappaB activation in children with congenital heart disease and the first evidence of myocardial nuclear factor kappaB translocation in human hearts before explant for transplantation. Furthermore, these data suggest that, similar to adults with advanced congestive heart failure, the myocardial inflammatory cascade may contribute to the pathophysiology of congenital heart disease in infants and children. PMID- 11940754 TI - Unexpected high risk of contamination with staphylococci species attributable to standard preparation of syringes for continuous intravenous drug administration in a simulation model in intensive care units. AB - OBJECTIVE: To determine the risk of bacterial contamination of the infusate in a simulation model of syringes prepared for continuous intravenous drug administration by intensive care unit nurses. Widely accepted standard procedures in the intensive care unit were compared with syringes prepared by pharmaceutical technicians working under standard aseptic conditions according to national guidelines. DESIGN: Prospective study. SETTING: Intensive care units of four general teaching hospitals and two university hospitals. SUBJECTS: We examined 650 syringes prepared from 10-mL ampules and 100 syringes prepared from rubber compound-capped 50-mL vials by intensive care unit nurses of six hospitals. Also, 100 syringes from 10-mL ampules and 100 syringes from rubber-compound-capped 50 mL vials were prepared by pharmaceutical technicians. INTERVENTIONS: Turbidity of syringes after culturing for 7 days at 37 degrees C was used as the criterion for possible bacterial contamination, which was proved with subsequent Gram staining. MEASUREMENTS AND MAIN RESULTS: A median contamination rate of 22% (range, 7% to 44%) was observed for the syringes prepared from 10-mL ampules by intensive care unit nurses, compared with only 1% for the syringes prepared from ampules by technicians (p <.001). In >75% of all contaminated syringes, Gram-positive cocci were identified. At least 12% of all prepared syringes proved to be contaminated with staphylococci species. The contamination rate of syringes prepared from vials was much lower: 2% in the intensive care unit and 0% at the department of clinical pharmacy. CONCLUSIONS: In the intensive care unit, standard procedures for preparing syringes for intravenous administration of drugs lack vigorous aseptic precautions, leading to a high contamination rate of the infusate. This risk is increased when ampules instead of 50 mL-vials are used to prepare the syringes. PMID- 11940755 TI - Increased bone resorption in the critically ill: association with sepsis and increased nitric oxide production. AB - OBJECTIVE: Cytokines stimulate nitric oxide production in bone, and high concentrations of cytokine-induced nitric oxide inhibit bone resorption in vitro. This has led to the suggestion that nitric oxide may protect against bone loss in inflammatory and infectious diseases. In this study, we sought to determine whether nitric oxide generated as the result of sepsis was associated with suppression of bone resorption in vivo. DESIGN: Observational study. SETTING: Adult intensive care unit of a university hospital. PATIENTS: We studied 20 consecutive patients who had been admitted to the intensive care unit because of sepsis and three who had been admitted because of trauma. Controls were 29 patients with noninflammatory musculoskeletal conditions. INTERVENTIONS: Standard clinical care. MEASUREMENTS AND MAIN RESULTS: Bone resorption was assessed by measurement of urinary pyridinoline and deoxypyridinoline as a ratio to urinary creatinine. Nitric oxide production was assessed by measuring the ratio of the nitric oxide breakdown products nitrate and nitrite to urinary creatinine. Urinary nitrate and nitrite/creatinine values were significantly higher in intensive care patients with sepsis (mean +/- sem, 0.164 +/- 0.053 micromol/mmol) than in intensive care patients with trauma (0.066 +/- 0.008) and controls (0.079 +/- 0.007; p =.007 between groups). Urinary pyridinoline/creatinine values were increased in intensive care patients with sepsis (553.8 +/- 193 nmol/mmol) and trauma (238 +/- 32) compared with controls (44.7 +/- 2.6; p =.001 between groups), and similar differences between the groups were observed for deoxypyridinoline/creatinine values: intensive care patients with sepsis, 86.4 +/ 24.0; intensive care patients with trauma, 46 +/- 4.2; and controls, 10.3 +/- 0.7 (p =.001). CONCLUSIONS: Critically ill patients with sepsis have increased nitric oxide production and increased bone resorption, whereas trauma patients have increased bone resorption in the presence of normal nitric oxide production. High concentrations of nitric oxide generated during the course of infection do not afford significant protection against accelerated bone resorption. PMID- 11940757 TI - Frequency and determinants of drug administration errors in the intensive care unit. AB - OBJECTIVE: The study aimed to identify both the frequency and the determinants of drug administration errors in the intensive care unit. DESIGN: Administration errors were detected by using the disguised-observation technique (observation of medication administrations by nurses, without revealing the aim of this observation to the nurses). SETTING: Two Dutch hospitals. PATIENTS: The drug administrations to patients in the intensive care units of two Dutch hospitals were observed during five consecutive days. INTERVENTIONS: None. MEASUREMENTS AND MAIN RESULTS: A total of 233 medications for 24 patients were observed to be administered (whether ordered or not) or were observed to be omitted. When wrong time errors were included, 104 administrations with at least one error were observed (frequency, 44.6%), and when they were excluded, 77 administrations with at least one error were observed (frequency, 33.0%). When we included wrong time errors, day of the week (Monday, odds ratio [OR] 2.69, confidence interval [CI] 1.42-5.10), time of day (6-10 pm, OR 0.28, CI 0.10-0.78), and drug class (gastrointestinal, OR 2.94, CI 1.48-5.85; blood, OR 0.12, CI 0.03-0.54; and cardiovascular, OR 0.38, CI,0.16-0.90) were associated with the occurrence of errors. When we excluded wrong time errors, day of the week (Monday, OR 3.14, CI 1.66-5.94), drug class (gastrointestinal, OR 3.47, CI 1.76-6.82; blood, OR 0.21, CI 0.05-0.91; and respiratory, OR 0.22, CI 0.08-0.60), and route of administration (oral by gastric tube, OR 5.60, CI 1.70-18.49) were associated with the occurrence of errors. In the hospital without full-time specialized intensive care physicians (which also lacks pharmacy-provided protocols for the preparation of parenteral drugs), more administration errors occurred, both when we included (OR 5.45, CI 3.04-9.78) and excluded wrong time errors (OR 4.22, CI 2.36-7.54). CONCLUSIONS: Efforts to reduce drug administration errors in the intensive care unit should be aimed at the risk factors we identified in this study. Especially, focusing on system differences between the two intensive care units (e.g., presence or absence of full-time specialized intensive care physicians, presence or absence of protocols for the preparation of all parenteral drugs) may help reduce suboptimal drug administration. PMID- 11940756 TI - Inhibition of p38 mitogen-activated protein kinase: dose-dependent suppression of leukocyte and endothelial response after endotoxin challenge in humans. AB - OBJECTIVE: We studied the activity of a single oral dose of RWJ-67657, a synthetic p38 mitogen-activated protein kinase inhibitor, in preventing dual leukocyte/endothelial activation after endotoxin infusion in healthy volunteers. DESIGN: Prospective placebo-controlled study. SETTING: Intensive care unit at a university medical center. SUBJECTS: Twenty-one healthy male volunteers. INTERVENTIONS: Endotoxin (4 ng/kg) as a 1-min infusion. According to randomization, the volunteers received placebo (n = 6) or 1400 mg (n = 4), 700 mg (n = 6), or 350 mg (n = 5) of RWJ-67657. MEASUREMENTS AND MAIN RESULTS: Neutrophil activation was investigated by analyzing the extent of membrane expression of adhesion markers by calibrated flow cytometry. Circulating intercellular adhesion molecule-1 and E-selectin were measured by enzyme-linked immunosorbent assays. The endotoxin-induced shedding of L-selectin was diminished in a dose-dependent manner (p <.0001). High-dose RWJ-67657 prevented up regulation of the integrins CD11b (p <.01) and CD 66b (p <.01) on neutrophils. The endotoxin-induced increase in circulating intercellular adhesion molecule-1 and circulation E-selectin was almost completely prevented by high-dose RWJ 67657. CONCLUSION: A single oral dose of RWJ-67657 prevented neutrophil and endothelial activation after endotoxin infusion. PMID- 11940758 TI - Early albumin infusion improves global and local hemodynamics and reduces inflammatory response in hemorrhagic shock. AB - OBJECTIVE: To evaluate the effects of an early, short-term albumin infusion on mesenteric microcirculation and global hemodynamics in hemorrhagic shock. DESIGN: A prospective, randomized study. SETTING: Animal laboratory at a university medical clinic. SUBJECTS: Seventeen Sprague-Dawley rats weighing 250-400 g. INTERVENTIONS: The rats underwent median laparotomy and exteriorization of an ileal loop for intravital microscopy of the mesenteric microcirculation. Volume controlled hemorrhagic shock was provoked by arterial blood withdrawal (2.5 mL/100 g body weight for 60 mins), followed by a 4-hr reperfusion period. Albumin (20%) or 0.9% NaCl was administered intravenously as a continuous infusion for 30 mins at the beginning of reperfusion. Reperfusion time mimicked a "prehospital" phase of 30 mins followed by a quasi "in-hospital" phase of 3.5 hrs. The "in hospital" phase in both groups was initiated by substitution of blood followed by reperfusion with normal saline. MEASUREMENTS AND MAIN RESULTS: Central hemodynamics, mesenteric microcirculation, and arterial blood gas parameters were monitored before, during, and 60 mins after hemorrhagic shock, and for a 240-min follow-up period after initiation of reperfusion. Application of albumin markedly reduced rolling and adherent leukocytes, maximum velocity, and shear rate in the mesenteric microcirculation. Later, after improvement of mesenteric microcirculation, an intermittent increase of central venous pressure and abdominal blood flow and decrease of hematocrit was observed. CONCLUSIONS: Albumin treatment of hemorrhagic shock improves microcirculation and global hemodynamics and attenuates the inflammatory response to reperfusion. It may provide clinical benefit when applied at an early stage of reperfusion during hemorrhagic shock. PMID- 11940759 TI - Effects of manganese superoxide dismutase, when given after inhalation injury has been established. AB - OBJECTIVES: To determine whether treatment with manganese superoxide dismutase (MnSOD), given intravenously after inhalation injury has been established, improves oxygenation and lung fluid balance. DESIGN: Randomized, controlled intervention trial. SETTING: University research laboratory. SUBJECTS: Twenty four chronically instrumented awake ewes with lung lymph fistulas. INTERVENTIONS: After smoke inhalation with 48 breaths of cotton smoke, the animals were assigned randomly to a control group (n = 6) or a treatment group, receiving 1000 units of MnSOD/kg (n = 6), 3000 units of MnSOD/kg (n = 6), or 9000 units of MnSOD/kg (n = 6) intravenously 1 hr after smoke inhalation. MEASUREMENTS AND MAIN RESULTS: Different from the other three groups, in the group that received 3000 units of MnSOD, cardiac output and Pao2/Fio2 ratio did not significantly decrease throughout the experimental period. Apart from higher oxygen consumption in the group receiving 3000 units of MnSOD 24 hrs after smoke inhalation (263 +/- 44 mL/min vs. 182 +/- 36 mL/min; p < 0.05), no significant differences between treatment groups and control group were observed. CONCLUSIONS: Treatment with MnSOD given after smoke inhalation seems to be less effective then pretreatment with MnSOD, which was reported in previous studies to reduce the degree of inhalation injury. PMID- 11940760 TI - Reduced rate of bacterial translocation and improved variables of natural killer cell and T-cell activity in rats surviving controlled hemorrhagic shock and treated with hypertonic saline. AB - OBJECTIVE: To examine the effect of hypertonic saline on bacterial translocation and the number and function of natural killer and T cells in controlled hemorrhagic shock. DESIGN: Randomized controlled study. Duration of follow-up was 24 hrs. SETTING: University research laboratory. SUBJECTS: Adult male Sprague Dawley rats, weighing 310-390 g. INTERVENTIONS: Controlled hemorrhagic shock was induced by blood withdrawal (mean arterial pressure, 30-40 mm Hg) and maintained for 30 mins. The animals were randomly divided into three groups: group 1 (n = 10) was sham-operated, group 2 (n = 10) was untreated, and group 3 was treated with 5 mL/kg hypertonic saline (n = 10). The rats were killed after 24 hrs. MEASUREMENTS AND MAIN RESULTS: Infusion of hypertonic saline in group 3 was followed by reduced bacterial translocation rate (5.0 +/- 2.2% vs. 18.3 +/- 5.3%, p <.033). The total mass of bacteria isolated from hypertonic saline-treated animals with bacterial translocation was 7.8- to 10.4-fold less than that from untreated rats. Controlled hemorrhagic shock resulted in a low percentage of CD4+ cells in blood (35.2 +/- 3.9%, p <.05) and lymph nodes (44.4 +/- 4.5%, p <.05) and depressed CD4 expression on blood (82 +/- 13 arbitrary units, p <.005) and lymph node (168 +/- 24 arbitrary units, p <.03) cells. A compensatory mobilization of NKR-P1+ cells from lymph nodes (8.6 +/- 2.3%, p <.05) to blood (21.2 +/- 5.2%, p <.01) with down-regulated NKR-P1 expression on blood cells (59 +/- 10 arbitrary units, p <.005) was observed. Natural killer cell-mediated cytotoxicity was decreased (67.9 +/- 9.7%, p <.05). Hypertonic saline treatment greatly stimulated CD4 expression on blood (419 +/- 113 arbitrary units, p <.005) and lymph node (553 +/- 115 arbitrary units, p <.03) cells. Also, normalization of NKR-P1 expression (160 +/- 19 arbitrary units, p <.005) and restoration of natural killer cell-mediated cytotoxicity to near normal values (88.6 +/- 7.4%, p <.05) were demonstrated. CONCLUSION: Controlled hemorrhagic shock was accompanied by CD4+ cells suppression and excessive recruitment of natural killer cells with abnormally low NKR-P1 expression and suppressed cytolytic activity into circulation. Infusion of hypertonic saline reversed these changes and reduced bacterial translocation. PMID- 11940761 TI - Effects of inhaled nitric oxide in a mouse model of sepsis-induced acute lung injury. AB - OBJECTIVE: Although inhaled nitric oxide transiently improves oxygenation in patients with acute lung injury, it has not affected clinical outcomes. As well, the effects of inhaled nitric oxide on the pathophysiologic features of acute lung injury have not been well defined. Therefore, we assessed the effects of inhaled nitric oxide on the degree of pulmonary inflammation and injury in a mouse model of sepsis-induced acute lung injury. DESIGN: Randomized, controlled animal study. SETTING: Research laboratory of an academic institution. SUBJECTS: Male C57Bl/6 mice. INTERVENTIONS: Sepsis was induced by cecal ligation and perforation. At the time of surgery, septic and naive mice were randomized to exposure to either 40 ppm inhaled nitric oxide or room air for 24 hrs before they were killed. MEASUREMENTS AND MAIN RESULTS: Sepsis-induced acute lung injury was characterized by increased pulmonary myeloperoxidase (68 +/- 13 vs. 13 +/- 3 mU/mg protein in naive mice, p <.01), pulmonary 8-isoprostane content (627 +/- 51 vs. 88 +/- 20 pg/mg protein in naive mice, p <.01), and protein in bronchoalveolar lavage fluid (p <.05). Inhaled nitric oxide exposure in septic mice completely abrogated the septic increases in myeloperoxidase activity (p <.05) and pulmonary 8-isoprostane content (p <.05) but had no effect on bronchoalveolar lavage protein. The induction of sepsis also was associated with an increase in pulmonary inducible NO synthase activity (2.8 +/- 0.5 vs. 0.4 +/- 0.1 pmol small middle dotmin-1 small middle dotmg-1 protein in naive mice, p <.05), and inhaled nitric oxide attenuated this increase in pulmonary inducible NO synthase activity (p <.05). CONCLUSIONS: Exposure to inhaled nitric oxide early in the course of sepsis-induced acute lung injury is associated with reduced pulmonary leukocyte infiltration and less oxidative injury. Decreased lung inflammation and injury with inhaled nitric oxide is associated with decreased pulmonary inducible NO synthase activity. Therefore, inhaled NO may have greater clinical benefit if administered earlier in the natural history of acute lung injury in patients. PMID- 11940762 TI - Importance of hypoxic vasoconstriction in maintaining oxygenation during acute lung injury. AB - OBJECTIVE: To investigate the role of hypoxic pulmonary vasoconstriction in the intrapulmonary blood flow redistribution and gas exchange protection during oleic acid acute lung injury. DESIGN: Prospective, controlled animal study. SETTING: Research laboratory of an academic institution. SUBJECTS: Three groups of five mongrel dogs. INTERVENTIONS: Induction of acute lung injury by 0.08 mL/kg oleic acid intravenously. Hypoxic pulmonary vasoconstriction inhibition by Escherichia coli endotoxin microdose (15 microg/kg) pretreatment or by metabolic alkalosis (pH 7.60). MEASUREMENTS AND MAIN RESULTS: Pulmonary arterial and venous resistances were determined by flow-pressure curves and by capillary pressure estimation. Regional lung water and pulmonary blood flow were assessed by positron emission tomography. Oleic acid alone increased the arterial and venous resistances, redistributed blood flow away from edematous areas, and decreased the Pao2 from 507 +/- 16 to 373 +/- 60 torr. on Fio2 1.0 and positive end expiratory pressure 5 cm H2O. Endotoxin pretreatment inhibited the increase in arterial resistance, suppressed the redistribution, and decreased the Pao2 to 105 +/- 22 torr. Alkalosis inhibited the increase in arterial and venous resistances, suppressed the redistribution, and decreased the Pao2 to 63 +/- 12 torr. Reversal of the alkalosis increased the arterial and venous resistances, restored the perfusion redistribution, and improved the Pao2 to 372 +/- 63 torr. Changes in blood gases conformed to predictions of a computer lung model in which hypoxic pulmonary vasoconstriction was suppressed by endotoxin and alkalosis. CONCLUSIONS: We conclude that in oleic acid-induced lung injury, a) pulmonary hypertension results from increases in both arterial and venous resistances; b) the increase in arterial resistance is the primary mechanism responsible for the perfusion redistribution and the gas exchange protection; and c) the increase in arterial resistance is most consistent with hypoxic pulmonary vasoconstriction. PMID- 11940763 TI - Dopexamine but not dopamine increases gastric mucosal oxygenation during mechanical ventilation in dogs. AB - OBJECTIVE: To compare the effects of dopamine and dopexamine on gastric mucosal oxygenation during mechanical ventilation without and with positive end expiratory airway pressure (PEEP) and after compensation of the PEEP-induced hemodynamic suppression. DESIGN: Randomized controlled animal study. SETTING: University research department of experimental anesthesiology. SUBJECTS: Ten anesthetized dogs with chronically implanted ultrasound flow probes around the pulmonary artery for continuous measurement of cardiac output. INTERVENTIONS: On different days, the dogs randomly received dopamine (2.5 and 5.0 microg x kg(-1) x min(-1), n = 10), dopexamine (0.5 and 1.0 microg x kg(-1) x min(-1)) without (n = 8) or with pretreatment with a selective beta2-adrenoceptor antagonist (ICI 118,551, n = 7), or saline (control, n = 7). To simulate common clinical situations, these interventions were performed during different ventilation modes: during mechanical ventilation without and with high levels of PEEP, and after compensation of the PEEP-induced systemic hemodynamic suppression by titrated volume resuscitation with hydroxyethyl starch. MEASUREMENTS AND MAIN RESULTS: We continuously measured microvascular hemoglobin saturation (mu-Hbo2) by light-guide spectrophotometry in the gastric mucosa. Dopexamine, but not dopamine, significantly increased gastric mucosal mu-Hbo2 by about 20%, regardless of the dose and the ventilation mode. Both catecholamines dose dependently increased cardiac output and oxygen delivery by up to 75% without effects on systemic oxygen saturation. The effects of dopexamine on mu-Hbo2 as well as on cardiac output and oxygen delivery were prevented by selective beta2 adrenoceptor-blockade. CONCLUSIONS: Dopexamine but not dopamine improved gastric mucosal oxygenation in dogs. This effect was independent of the dosage and the ventilation mode. Thus, dopexamine may reverse a decrease in splanchnic oxygenation induced by ventilation with PEEP. The dopexamine-induced increase in gastric mucosal oxygenation was mediated by beta2-adrenoceptors, which explains the superior effects of dopexamine to dopamine on mu-Hbo2. The regional effects of both catecholamines were not mirrored by systemic hemodynamics. PMID- 11940764 TI - Preservation of splenic immune functions by female sex hormones after trauma hemorrhage. AB - OBJECTIVES: Immune functions are markedly depressed in males but not in proestrus females after trauma-hemorrhage. Nonetheless, it is unclear what role sex steroids play in the maintenance of immune function in females after trauma hemorrhage. DESIGN: Prospective, controlled animal study. SETTING: University research laboratory. SUBJECTS: Eight-week-old female CBA/J mice. INTERVENTIONS: Mice underwent sham-ovariectomy or ovariectomy. Two weeks thereafter, ovariectomized and proestrus sham-ovariectomized mice were subjected to laparotomy (i.e., soft tissue trauma) and hemorrhagic shock (35 +/- 5 mm Hg for 90 mins, resuscitated) or sham operation. Splenocyte proliferation and interleukin-2, interleukin-3, and interferon-gamma release were determined at 2 hrs after trauma-hemorrhage. MEASUREMENTS AND MAIN RESULTS: These immune functional capacities were maintained in proestrus sham-ovariectomized mice after trauma-hemorrhage, whereas they were suppressed in ovariectomized mice subjected to trauma-hemorrhage. 17beta-Estradiol in vitro had no effect on splenocyte functions in proestrus sham-ovariectomized females; however, addition of 17beta estradiol to splenocytes from ovariectomized females subjected to trauma hemorrhage normalized immune functional capacities. CONCLUSIONS: These findings suggest that elevated circulating 17beta-estradiol in proestrus females plays a direct role in the maintenance of immunocompetence after trauma-hemorrhage. PMID- 11940765 TI - Modification of alpha1 -adrenoceptors by peroxynitrite as a possible mechanism of systemic hypotension in sepsis. AB - OBJECTIVE: It is well known that nitric oxide synthase is induced by endotoxin or inflammatory cytokines, and consequently large amounts of nitric oxide cause vascular hyporeactivity to vasoconstrictor agents and myocardial dysfunction, hence hypotension. However, there is considerable controversy as to whether these pathologic cardiovascular features are mediated directly by nitric oxide or also through the formation of secondary reaction products such as peroxynitrite (ONOO 1). Our objective was to investigate inhibitory effects of ONOO-1 on alpha1 adrenoceptors. DESIGN: Prospective, controlled, in vitro, laboratory study. SETTING: Laboratory of a health sciences university. SUBJECTS: Chinese hamster ovary cells that expressed the human recombinant alpha1a-, alpha1b-, or alpha1d adrenoceptors, rat aorta strips. INTERVENTIONS: Binding experiments of [3H]prazosin were done in the Chinese hamster ovary cell membranes pretreated with 100 microM to 3 mM ONOO-1. Displacement experiments with noradrenaline or 3 nitro-l-tyrosine also were conducted. Mobilization of intracellular Ca2+ evoked by 1 nM to 10 microM noradrenaline was monitored in a fluorescence spectrophotometer with dual excitation at 340 nm/380 nm and emission at 500 nm in fura-2/AM-loaded Chinese hamster ovary cells. Contractile force produced by noradrenaline was monitored in rat aorta strips that have alpha1a- and alpha1d adrenoceptors, pretreated with 1 mM ONOO-1. Either 0.3 N NaOH or the decomposed ONOO-1 was used as the control. MEASUREMENTS AND MAIN RESULTS: The specific binding of [3H]prazosin to alpha1a- and alpha1d-adrenoceptor was inhibited by ONOO-1 in a concentration-dependent manner. We found that 3 mM ONOO-1 decreased maximum binding sites by 40% to 50% in alpha1a- and alpha1d-adrenoceptors. Binding affinities for prazosin and noradrenaline were not affected by 1 mM ONOO 1 in all subtypes. We found that 3-nitro-l-tyrosine did not affect the prazosin binding to three adrenoceptor subtypes. Noradrenaline increased intracellular Ca2+ concentration ([Ca2+]i) concentration-dependently, which was inhibited by ONOO-1 in alpha1a- and alpha1d-adrenoceptors. ONOO-1 had no effect on alpha1b adrenoceptor. Contractile force produced by noradrenaline decreased significantly in aorta strips pretreated with ONOO-1. CONCLUSION: ONOO-1 reduces the binding capacity of alpha1a- and alpha1d- but not alpha1b-adrenoceptors without changing the affinities. Treatment with ONOO-1 attenuates noradrenaline-stimulated increase in [Ca2+]i in alpha1a- and alpha1d-adrenoceptors but not in alpha1b adrenoceptor. ONOO-1 also weakens noradrenaline-induced contractions in rat aorta that has alpha1a- and alpha1d-adrenoceptors. Cardiovascular hyporeactivity to catecholamines in septic shock may be caused in part by the inactivation of alpha adrenoceptors by ONOO-1. PMID- 11940766 TI - Citrate anticoagulation in a piglet model of pediatric continuous renal replacement therapy. AB - OBJECTIVE: To develop pediatric guidelines for the use of citrate as a regional anticoagulant for continuous renal replacement therapy (CRRT) using a neonatal piglet model. DESIGN: Prospective observational study. SETTING: Animal laboratory in the research center of a tertiary-level children's hospital. SUBJECTS: Ten neonatal piglets. INTERVENTIONS AND MEASUREMENTS: Using a venovenous CRRT circuit and filter, we randomly altered the filter blood flow rate, replacement flow rate, and citrate flow rate over conventional pediatric ranges. Measured end points were prefilter serum ionized calcium and citrate levels. MAIN RESULTS: A prefilter serum citrate concentration of 6 mmol/L is required to maintain the prefilter ionized calcium < or =0.4 mmol/L. Using multiple regression analysis on collected data, we derived a formula to predict prefilter serum citrate for combinations of replacement flow rate, blood flow rate, and citrate flow rate. CONCLUSIONS: The available literature and our past experience indicate that a prefilter ionized calcium < or =0.4 mmol/L is required to anticoagulate a CRRT circuit; a prefilter serum citrate concentration of 6 mmol/L is required to achieve this. Our multiple regression analysis can be expressed graphically to allow easy calculation of the required citrate flow rate, given the knowledge of the replacement flow rate and blood flow rate. Our results provide the first guidelines for the use of citrate as a regional anticoagulant in a pediatric-size model of CRRT. PMID- 11940767 TI - Effects of posttreatment with propofol on mortality and cytokine responses to endotoxin-induced shock in rats. AB - OBJECTIVE: To clarify the effects of posttreatment with propofol administration on mortality rate and cytokine responses to endotoxin-induced shock in rats. DESIGN: Randomized prospective laboratory study. SETTING: University laboratory. SUBJECTS: Thirty-three male rats. INTERVENTIONS: Animals were randomly assigned to one of three groups (n = 11 per group): a) endotoxemic group, receiving intravenous Escherichia coli endotoxin (20 mg/kg over 2 mins); b) early posttreatment group, treated identically to the endotoxemic group with the additional administration of propofol (10 mg/kg bolus, followed by infusion at 10 mg x kg(-1) x hr(-1)) 1 hr after the injection of endotoxin; and c) late posttreatment group, treated identically to the endotoxemic group with the additional administration of propofol (10 mg/kg bolus, followed by infusion at 10 mg x kg(-1) x hr(-1)) 2 hrs after the injection of endotoxin. MEASUREMENTS AND MAIN RESULTS: Hemodynamics and arterial blood gases were recorded, and mortality rate and plasma cytokine concentrations were calculated for the 5-hr observation. The mortality rate 5 hrs after endotoxin injection was 73% for the endotoxic, 9% for the early posttreatment, and 36% for the late posttreatment groups. The mortality rate for the early posttreatment group was significantly lower than that for the other groups. The increases in plasma cytokine (tumor necrosis factor-alpha and interleukin-6 and -10) concentrations were less for the early posttreatment group than the other two groups. CONCLUSIONS: The early posttreatment of propofol after endotoxin injection drastically reduced the mortality rate of rats and attenuated their cytokine responses. Moreover, propofol attenuated the production of tumor necrosis factor-alpha. These findings suggest that propofol administration may be beneficial during sepsis. PMID- 11940768 TI - Fibrin sheath enhances central venous catheter infection. AB - OBJECTIVE: To determine whether fibrin-coated central venous catheters have a higher infection rate, and spawn more septic emboli, than uncoated catheters after exposure to bacteremia. DESIGN: Animal study comparing catheter infection and blood cultures of fibrin-coated and uncoated catheters exposed to bacteremia. SETTING: Animal laboratory. SUBJECTS: Adult male Sprague-Dawley rats. INTERVENTIONS: A total of 210 rats had catheters placed with the proximal end buried subcutaneously. Rats were divided into three groups: tail vein bacterial injection on day 0 (no fibrin group) or on day 10 (fibrin group), or no injection/saline injection (control, n = 40). Bacterial injections were 1 x 108 colony forming units of either Staphylococcus epidermidis (n = 100) or Enterobacter cloacae (n = 60). Animals were killed 3 days after injection. Blood cultures were obtained via cardiac puncture, and catheters were removed via the chest. Half of the catheter was rolled onto agar and the other half was placed in trypticase soy broth. Plates and broth were incubated at 37 degrees C for 48 hrs. The presence of >15 colonies on roll plates, or growth in broth, was accepted as a positive sign of infection. Microscopy was performed on day 20-10 catheters. Thirty animals without catheters had bacterial injections and underwent blood culture 3 days after injection. MEASUREMENTS AND MAIN RESULTS: Catheter infection with S. epidermidis occurred in 32% of roll plates and 80% of broth from the fibrin group vs. 4% and 20% from the no fibrin group (p <.01 for each). Catheter infection with E. cloacae occurred in 50% of roll plates and 80% of broth from the fibrin group vs. 0% and 12% from the no fibrin group (p <.01 for each). Positive blood cultures occurred in 47 of 68 animals from the fibrin group vs. 8 of 68 from the no fibrin group (p <.01). Microscopy showed a fibrin sheath on 20 of 20 catheters. Without catheters, 30 of 30 blood cultures were negative. CONCLUSION: The fibrin sheath significantly enhanced catheter-related infection and persistent bacteremia. PMID- 11940769 TI - Effect of dexamethasone on the expression of brain-derived neurotrophic factor and neurotrophin-3 messenger ribonucleic acids after forebrain ischemia in the rat. AB - OBJECTIVE: To determine whether a large dose of dexamethasone affected brain damage induced by concurrent cerebral ischemia, we used in situ hybridization to examine the expression of brain-derived neurotrophic factor and neurotrophin-3 messenger ribonucleic acids (mRNAs) in rats with and without dexamethasone administration after transient forebrain ischemia. DESIGN: Prospective experimental study in rats. SETTING: Experimental laboratory in a teaching hospital and university. SUBJECTS: Eighty adult rats. INTERVENTIONS: Twenty minutes of transient forebrain ischemia was induced by occlusion of four vessels in lightly anesthetized rats. Thirty-six animals received dexamethasone (15 mg/kg, intraperitoneally) after initial reperfusion. Thirty-six dexamethasone control rats were injected with saline, and the remaining animals underwent sham surgery but no ischemia or dexamethasone. MEASUREMENTS AND MAIN RESULTS: Using in situ hybridization, we determined hippocampal brain-derived neurotrophic factor and neurotrophin-3 mRNA expression 2, 4, 6, 12, and 24 hrs and 2, 3, 4, and 7 days after brain ischemia. Additionally, hippocampal CA1 region cell death was measured with Nissl stains. Both brain-derived neurotrophic factor and neurotrophin-3 mRNA exhibited a biphasic response after ischemia. Brain-derived neurotrophic factor mRNA showed two peaks of 4.07-fold and 2.84-fold increases relative to sham operation at 6 hrs and 2 days, respectively. Neurotrophin-3 mRNA initially decreased to 59% of sham levels at 4 hrs and then increased to 146% at 3 days before it returned to basal levels. When the ischemic rats were treated with dexamethasone, the elevation of brain-derived neurotrophic factor mRNA and the reduction of neurotrophin-3 mRNA level were prevented within the first 24 hrs, and hippocampal CA1 neurons were protected from ischemia-induced cell loss 7 days after brain ischemia. The protein levels of both brain-derived neurotrophic factor and neurotrophin-3 in general correspond to the mRNA levels in the hippocampal region. CONCLUSIONS: Dexamethasone modulates the intriguing temporal and spatial expression of brain-derived neurotrophic factor and neurotrophin-3 that predominantly supports neuronal innervation at different times after brain ischemia and also may provide specific trophic support for various neurons in the central nervous system. PMID- 11940770 TI - Impact of a pediatric clinical pharmacist in the pediatric intensive care unit. AB - OBJECTIVE: To study the impact of a clinical pharmacist in a pediatric intensive care unit. The goals of the study were to determine the type and quantity of patient care interventions recommended by a clinical pharmacist and to specifically examine cost savings (or loss) that resulted from clinical pharmacist recommendations. DESIGN: A prospective case series. SETTING: Ten-bed pediatric intensive care unit in a university-affiliated children's hospital. PATIENTS: All patients admitted to the pediatric intensive care unit during the study period. INTERVENTIONS: None. MEASUREMENTS AND MAIN RESULTS: During the 24 wk study period, the pediatric clinical pharmacist documented all interventions that occurred during her shift. She rounded with the pediatric intensive care unit team approximately two times a week and reviewed medication lists daily. Drug acquisition costs were used to calculate drug cost savings. Demographic information was collected on all the patients in the pediatric intensive care unit during the study period. There were 35 recommendations per 100 patient days. The most common interventions were dosage changes (28%), drug information (26%), and miscellaneous information (22%). The average time spent per day by the clinical pharmacist in the pediatric intensive care unit was 0.73 hrs or 0.02 full-time equivalent. The total cost direct savings for the study period was $1,977. Extrapolated to direct cost savings per year, the total amount saved was $9,135/year or 0.15 full-time equivalent. Indirect savings from educational activities, avoidance of medication errors, and optimization of medical therapies represent an additional nonquantifiable amount. CONCLUSION: We conclude that a clinical pharmacist is an important and cost-effective member of the pediatric intensive care unit team. PMID- 11940771 TI - Benefits of early tracheostomy in severely burned children. AB - OBJECTIVE: The role of tracheostomy in burn patients is controversial. Previous studies, primarily in adults, suggested that severely burned patients with tracheostomies have a higher incidence of tracheostomy site infections, mortality, and pneumonia. The purpose of this study is to determine the safety and efficacy of early tracheostomy in severely burned children. DESIGN: Case series study analyzing mechanical ventilation and sedation requirements before and 24 hrs after tracheostomy. SETTING: Regional pediatric burn center. PATIENTS: All children admitted to a regional pediatric burn center requiring tracheostomy from March 1, 1998, to October 1, 2001. METHODS: Data were recorded on patients' demographics, extent of burn, presence of inhalation injury, and mortality. Mechanical ventilation variables measured pretracheostomy (pre) and posttracheostomy (post) and included mode of ventilation, ventilator settings, peak inspiratory pressures, and arterial blood gases (Pao2, Paco2, pH, and oxygen saturation). Calculated variables included compliance, Pao2:Fio2 ratio, and minute ventilation. Tracheostomy-related variables recorded included the interval to tracheostomy insertion, the duration of tracheostomy, and tracheostomy complications. MAIN RESULTS: A total of 38 patients (with a mean age of 4.7 +/- 0.6 yrs and a mean total body surface area involvement of 54% +/- 4%, 63% with inhalation injury) underwent tracheostomy a mean of 3.9 +/- 0.7 days after admission. Overall mortality was 21%. There were no tracheostomy site infections, tracheostomy-related deaths, or tracheal stenoses in survivors. Peak inspiratory pressures were lower after tracheostomy (30.4 +/- 1.4 [pre] vs. 27.6 +/- 1.5 cm H2O [post]; p <.05), ventilatory volumes were higher (190 +/- 22 mL [pre] vs. 225.5 +/- 25 [post]; p <.05), compliance improved (10.5 +/- 1.4 [pre] vs. 15.1 +/ 2.3 mL/cm H2O [post]; p <.05), and the Pao2:Fio2 ratio improved (300.6 +/- 20 [pre] vs. 348.6 +/- 16 [post]). There was no difference in oxygenation, ventilation, minute ventilation, or pH after tracheostomy. CONCLUSIONS: Early tracheostomy in severely burned children is safe and effective. It provides a secure airway and may result in improvement in ventilator management for these children. PMID- 11940772 TI - Corticosteroids in the treatment of severe mycoplasma encephalitis in children. AB - OBJECTIVE: To report an unusual case of mycoplasma-associated encephalitis that responded to corticosteroid therapy and relapsed after cessation. DESIGN: Clinical case report. SETTING: Tertiary care pediatric intensive care unit. PATIENT: A single patient admitted to the pediatric intensive care unit. INTERVENTIONS: Intravenous corticosteroids and antibiotics. MEASUREMENTS AND MAIN RESULTS: Resolution of neurologic symptoms with initial steroid therapy, relapse after withdrawal of steroids, and resolution again with re-institution of steroid therapy. CONCLUSIONS: Encephalitis is a well-recognized and potentially severe complication of mycoplasma infections. Treatment with corticosteroids has been proposed for this illness, but experience with this therapy is limited to case reports and small series; neither efficacy nor appropriate courses of treatment are well established. The relapsing course of this patient, along with a review of prior reported cases, suggests that corticosteroids may be beneficial for mycoplasma encephalitis, that moderate doses may be sufficient, and that consideration should be given to a prolonged tapering course when these medications are used for this illness. PMID- 11940773 TI - Percutaneous repair of punctured subclavian artery under fluoroscopy. PMID- 11940774 TI - Resolution of severe Donath-Landsteiner autoimmune hemolytic anemia temporally associated with institution of plasmapheresis. AB - OBJECTIVES: To report a case of severe postinfectious autoimmune hemolytic anemia (AIHA) owing to the Donath-Landsteiner (DL) antibody resolving with plasmapheresis, and to review the pathophysiology of this underrecognized cause of pediatric AIHA and its potential susceptibility to plasmapheresis therapy. DESIGN: Descriptive case report. SETTING: A pediatric intensive care unit in a university children's hospital. PATIENT: A 5-yr-old Hispanic female had gastroenteritis followed by progressive intravascular hemolysis, initially attributed to acute postinfectious cold hemagglutinin (immunoglobulin M) disease. INTERVENTION: With no slowing in the rate of hemolysis, a continued need for frequent transfusions, and a lack of response to corticosteroid and intravenous immunoglobulin therapy, a 3-day course of plasmapheresis was administered. MEASUREMENTS AND MAIN RESULTS: The patient presented to an emergency department with an initial hematocrit of 22%, which fell to 12% by hospital admission. She received nine transfusions over 7 days, with her hematocrit reaching a nadir of 11% on the 5th day of hospitalization. Once plasmapheresis was initiated, she required no further transfusion. Analysis of serum from initial presentation demonstrated biphasic hemolysis, confirming the presence of the DL antibody. CONCLUSIONS: In AIHA, in which the direct antiglobulin test detects primarily C3 rather than immunoglobulin G, especially in children, the DL antibody must be considered. Confirming the diagnosis rapidly may be critical, especially in cases of severe hemolysis, because this may help direct therapy. A low titer of DL antibody can mediate severe intravascular hemolysis given its propensity to sensitize, detach, and rebind erythrocytes with changes in temperature in the microcirculation. However, given the transient and relatively brief production of the DL antibody in postviral illness, early clearance of the offending antibody may be possible with plasmapheresis, without the expectation for significant rebound antibody production, potentially decreasing the length of hospital stay and the need for transfusions. PMID- 11940775 TI - Therapeutic trials for traumatic brain injury--a journey in progress. PMID- 11940776 TI - Symptom management in the intensive care unit: toward a more holistic approach. PMID- 11940777 TI - Assault or homicide: treating and letting die without consent. PMID- 11940778 TI - Neutropenia recovery: good news or siren signal? PMID- 11940779 TI - Early extubation after pediatric heart surgery: the future? PMID- 11940780 TI - Inflammatory mediators in congenital heart disease. PMID- 11940781 TI - Ghosts of studies past, present, and future: a cautionary tale. PMID- 11940782 TI - Medication errors: problems identified, but what is the solution? PMID- 11940783 TI - Inhaled nitric oxide: is systemic host defense at risk? PMID- 11940784 TI - The many faces of sepsis-induced vascular failure. PMID- 11940785 TI - Therapeutic sedation: has its time come? PMID- 11940786 TI - Should time from cardiac arrest until call to emergency medical services (EMS) be collected in EMS research? AB - Because of the difficulty and cost of collecting the time from collapse to placing the 911 call in instances of out-of-hospital cardiac arrest and because of the potential noise and bias that might be inherent in such data, a simulation study was conducted to quantitate the impact that such data might have on estimates of the relationship between time from collapse to defibrillation and probability of survival. In the absence of bias, an underestimate of the slope on the order of 20% to 30% might be expected. However, in the presence of bias, the impact on the slope estimate is unpredictable. The most likely bias would tend to cause an overestimate of the slope. It is suggested that unless the time from collapse to placing the 911 call can be obtained accurately and without bias, it is probably not worthwhile to do so. PMID- 11940787 TI - Sex-related differences in the presentation and outcome of out-of-hospital cardiopulmonary arrest: a multiyear, prospective, population-based study. AB - OBJECTIVE: To examine whether previously observed sex-related differences in coronary artery disease syndromes also apply to patients with out-of-hospital sudden cardiac arrest, a probable subset of patients with coronary artery disease who are easy to recognize and are treated in a standardized fashion. DESIGN: Prospective, population-based study conducted over a 6-yr period. SETTING: A large urban municipality (population, 1.1 million) served by a single emergency medical services system with centralized medical direction and standardized protocols. PATIENTS: All patients with out-of-hospital, nontraumatic, primary cardiac arrest. INTERVENTIONS: Standardized advanced cardiac life support protocols. MEASUREMENTS AND MAIN RESULTS: During the 6 yrs of the study, 4147 consecutive patients were studied, 42% of whom were women (p <.001). Although women were significantly older than men (mean age, 68.7 +/- 18 vs. 61.7 +/- 17 yrs; p =.001), there were no significant differences for the percentages of witnessed and unwitnessed arrests, response intervals, and the length and type of treatment provided. Although men were more likely to have ventricular fibrillation/ventricular tachycardia on presentation (41% vs. 30%), women had more asystole (8.8% vs. 7%) and (organized) pulseless electrical activity than men (24% vs. 18%; p <.001). Nevertheless, more women were resuscitated (13.5% vs. 10.7%; p =.005), particularly women with non-ventricular fibrillation/ventricular tachycardia presentation (12.6% vs. 9.6%; p <.02). These differences were more pronounced when controlling for age (95% confidence interval, 1.44 [1.25-1.74]). CONCLUSIONS: In cases of out-of-hospital sudden cardiac arrest, women have significantly better resuscitation rates than men, especially when controlling for age, particularly among women with non-ventricular fibrillation/ventricular tachycardia presentations. Additional studies are required to validate these observations, not only for long-term survival and external validity, but also for other potential genetic factors and potential discrepancies with other studies. PMID- 11940788 TI - Reconsideration of proximate Utstein-style end points. AB - The Utstein-style template defines core and supplementary data for reporting out of-hospital cardiac arrest information. The primary outcome statistic of the Utstein template is survival to hospital discharge (SHD). The SHD statistic is dependent on Utstein-defined out-of-hospital variables and multiple in-hospital variables that are undefined and uncontrolled. An example of one of these undefined in-hospital variables is the decision to place a patient on do-not resuscitate status. At our municipal teaching hospital, 418 patients who had out of-hospital cardiac arrest presented over a 4-yr period; 79 (19%; 95% confidence interval [CI], 15% to 23%) survived to hospital admission, with 54 (68%; 95% CI, 57% to 78%) subsequently being placed on do-not-resuscitate status. When patients on do-not-resuscitate status were included in the SHD calculation, the SHD rate was 5.3% (95% CI, 3.3% to 7.8%), and when patients on do-not-resuscitate status were excluded from the SHD calculation, the SHD rate was 6.1% (95% CI, 3.8% to 9.0%). These data show a relative 15% change in SHD resulting from a single in hospital variable. Cardiac arrest survivors represent a small proportion of a total population; therefore, large numbers of study subjects are required for a statistically significant interpretation of the SHD statistic. This requirement for large study populations has resulted in recent studies that report results by using end points proximate to SHD when assessing the effect of individual interventions. It is logical that success of a specific intervention should be determined by the ability of the intervention to accomplish its purpose rather than the ability to improve SHD that is dependent on multiple variables. Furthermore, because in-hospital care is not standardized and uncontrolled variables exist, the primary Utstein end point of SHD should be reconsidered when evaluating cardiac arrest interventions. PMID- 11940789 TI - Cerebral resuscitation potentials for cardiac arrest. AB - Permanent brain damage after cardiac arrest and resuscitation is determined by many factors, predominantly arrest (no-flow) time, cardiopulmonary resuscitation (low-flow) time, and temperature. Research since around 1970 into cardiopulmonary cerebral resuscitation has attempted to mitigate the postischemic-anoxic encephalopathy. These efforts' results have recently shown outcome benefits as documented in clinically relevant outcome models in dogs and in clinical trials. Pharmacologic strategies have so far yielded relatively disappointing results. In a recent exploration of 14 drugs in dogs, only the antioxidant tempol administered at the start of prolonged cardiac arrest improved functional outcome in dogs. Cerebral blood flow promotion by hypertensive reperfusion and hemodilution has resulted in improved outcome in dogs, and brief hypertension after restoration of spontaneous circulation is associated with improved outcome in patients. Postarrest hypercoagulability of blood seems to yield to therapeutic thrombolysis, which is associated with improved cerebral outcome in animals and patients. In a clinically relevant dog outcome model, mild postarrest cerebral hypothermia (34 degrees C), initiated with reperfusion and continued for 12 hrs, combined with cerebral blood-flow promotion increased from 5 to >10 mins the previously longest normothermic no-flow time that could be reversed to complete cerebral recovery. Mild hypothermia by surface cooling after prolonged cardiac arrest in patients has been found effective in recent clinical studies in Australia and Europe. Preliminary data on the recent randomized study in Europe have been reported. For presently unresuscitable cardiac arrests, research since the 1980s in dog outcome models of prolonged exsanguination cardiac arrest has culminated in brain and organism preservation during cardiac arrest (no-flow) durations of up to 90 mins, perhaps 120 mins, at a tympanic temperature of 10 degrees C and complete recovery of function and normal histology. This "suspended animation for delayed resuscitation" strategy includes use of an aortic flush of cold saline (or preservation solution) within the first 5 mins of no flow. This strategy should also be explored for the larger number of patients with unresuscitable out-of-hospital cardiac arrests. Suspended animation for prolonged preservation of viability could buy time for transport and repair during hypothermic no flow followed by resuscitation, or it could serve as a bridge to prolonged cardiopulmonary bypass. PMID- 11940794 TI - Augmentation of ventricular preload during treatment of cardiovascular collapse and cardiac arrest. AB - Despite aggressive measures for the treatment of cardiovascular collapse and cardiac arrest, the hypotension associated with these malignant processes usually leads to profound vital-organ ischemia and death. A fundamental therapeutic challenge of such life-threatening processes is the restoration of adequate blood flow to the heart and the brain. However, to maintain adequate forward blood flow out of the heart, venous blood return must be drawn back into the heart. With the exception of administration of exogenous fluid replacement, there are limited ways to enhance blood flow back to the heart during prolonged hypotension. This article describes the potential value of a new impedance threshold valve for the treatment of cardiac arrest and hypotension. The valve was designed to create a vacuum within the thorax during the decompression phase of cardiopulmonary resuscitation or during inhalation. By transiently blocking inspiratory gas exchange during the decompression phase of cardiopulmonary resuscitation, after phrenic nerve-stimulated gasping, or during spontaneous ventilation, the impedance-valve concept may have clinical value in the treatment of patients in cardiac arrest, hemorrhagic shock, and cardiovascular collapse secondary to a number of life-threatening clinical processes. PMID- 11940795 TI - Myocardial protection during ventricular fibrillation by inhibition of the sodium hydrogen exchanger isoform-1. AB - Activation of the sarcolemmal sodium-hydrogen exchanger isoform-1 (NHE-1) in response to the intense intracellular acidosis that develops during ischemia has been identified as an important mechanism of myocardial cell injury. NHE-1 inhibition in the quiescent (nonfibrillating) heart ameliorates functional manifestation of ischemia and reperfusion injury. We investigated in isolated heart and intact rat models of ventricular fibrillation whether NHE-1 inhibition, by using the selective inhibitor cariporide, could ameliorate myocardial abnormalities that develop during ventricular fibrillation and limit resuscitability and survival. In the isolated rat heart, cariporide significantly reduced the magnitude of ischemic contracture during ventricular fibrillation and the accompanying increases in coronary vascular resistance. Hearts that had received cariporide during ventricular fibrillation had no diastolic dysfunction after resuscitation and recovered their systolic function earlier. In intact rats, cariporide given immediately before starting chest compression allowed generation of a coronary perfusion pressure and end-tidal Pco2 comparable with control rats but with significantly less depth of compression. Cariporide had an unprecedented effect in this rat model, prompting spontaneous defibrillation after approximately 8 mins of chest compression. After resuscitation, rats treated with cariporide had significantly less ventricular ectopic activity, better hemodynamic function, and higher survival rates (22 of 24 [94%] vs. 15 of 24 [63%] in control rats, p <.05). We conclude that NHE-1 inhibition may represent a novel and highly effective form of treatment for resuscitation from ventricular fibrillation. PMID- 11940798 TI - Convulsive Therapy. PMID- 11940799 TI - A Prospective Follow-Up Study of Cognitive Functions After Electroconvulsive Therapy. AB - The authors studied primarily nonmemory cognitive functioning in a sample of 13 melancholic patients tested prior to electroconvulsive therapy (ECT) and at various intervals during a 1-2 year post-ECT follow-up period. Compared with 13 age-matched normal controls, the patients performed significantly worse at baseline and immediately after the sixth ECT, but were not significantly different at the 30-day, 6-month, and 1-2 year assessments. At 1-2 years post ECT, cognitive impairment among patients was substantially and significantly less than observed pre-ECT. PMID- 11940796 TI - Preconditioning and postresuscitation injury. AB - A major challenge in improving cardiac arrest survival is organ injury that occurs after the return of spontaneous circulation. This postresuscitation injury may result in as many as 90% of such patients not surviving to hospital discharge. Preconditioning, an adaptive physiologic response found in multiple organs and species, may help protect against such injury of ischemic tissue when reperfused at the return of spontaneous circulation. A better understanding of how preconditioning may alter postresuscitation injury is important for two major reasons. First, it is one of the most protective adaptations currently known in nature that attenuates ischemia-reperfusion injury. Pharmacologic and nonpharmacologic means to quickly trigger and perhaps augment this response have the potential to greatly improve survival from the global ischemia of cardiac arrest. Second, potential targets of preconditioning-such as the adenosine triphosphate-sensitive potassium channel and NAD(P)H oxidases-likely play important roles in the postresuscitation phase of cardiac arrest, and their modification may be important components of future treatment for patients with return of spontaneous circulation. The evidence for postresuscitation injury at the cellular level and its modification by preconditioning are discussed. PMID- 11940800 TI - Outcome Following Electroconvulsive Therapy: A Comparison of Primary and Secondary Depression. AB - In a consecutive series of 117 depressed patients referred for electroconvulsive therapy, the 10 with secondary depression had significantly poorer outcomes according to three independently assessed measures-Hamilton rating scale score at discharge, global rating at discharge, and mean depressive symptom score during a 6-month follow-up. PMID- 11940801 TI - Electroconvulsive Therapy-Induced Cortisol Release: Changes with Depressive State. AB - With administration of 2 mg of dexamethasone 9 h before, serum cortisol levels were determined for the times just before and 30 min after the first and last electroconvulsive therapies (ECTs) of a series of at least six sessions. Twelve drug-free patients with melancholic or psychotic depression completed the study. Response to treatment was taken as a fall in the value of the Carroll rating scale. The average cortisol elevation over baseline was by 575% with the first ECT, 10 times larger than other studies in the literature that were done without dexamethasone. With the final ECT the elevation was lower (p < 0.01), averaging 181%. Post-ECT cortisol levels (and elevations over baseline) were lower after the final ECT than after the initial ECT, with 10 of the 11 patients who responded to this treatment (p < 0.005 vs. 50%). Consistent with this, the lone nonresponder showed a large increase in serum cortisol level and elevation over baseline from the first to the last treatment. The one responder who went against this trend had an unusually high baseline cortisol level, 24.1 &mgr;g/dl, which can be considered to disqualify his inclusion. The near-unanimous fall of post ECT cortisol levels with remission suggests that defects in cortisol regulation are much more prevalent (p < 0.01) than those detected by the dexamethasone suppression test (36% in this sample). PMID- 11940802 TI - Total Energy and Rate of Application as Measures of the Electroconvulsive Therapy Stimulus. AB - This study evaluated the utility of two measures of the electroconvulsive therapy (ECT) stimulus: Total energy and rate of application of energy. Data for more than 2,000 trials drawn from six previous studies were divided into success and failure groups based on the production of a satisfactory clinical convulsion. The distributions and descriptive statistics for both measures and all studies were presented. The range for energy was 4-80+ J, with both successes and failures proportionately distributed across the entire range. Rate varied from 1 to 16+ J/s, also with proportional distribution. The presence of a different interpulse interval in some of the studies allowed a demonstration showing that the rate measure was susceptible to considerable influence by this stimulus characteristic. As clinical considerations reduce the number of observations made at the higher energy levels, it was concluded that the influence of both energy and rate of application was underestimated substantially. Both energy and rate supply useful information about the ECT stimulus, but are much too limited to serve as adequate descriptors. Clinical illustrations are provided. PMID- 11940803 TI - Detection of Prolonged Seizures During Electroconvulsive Therapy: A Comparison of Electroencephalogram and Cuff Monitoring. AB - In a retrospective study of 1,223 electroconvulsive therapy seizures, the incidence of prolonged seizures (those with durations greater than 120 s) was 4.25%. Prolonged seizures are rarely reported. The author speculates that the higher incidence reported here may be secondary to the higher succinylcholine dosage (0.75-1.0 mg/kg) employed at this center. Electroencephalogram (EEG) and blood pressure cuff monitoring techniques are compared. EEG monitoring is found superior to cuff monitoring for the detection of prolonged seizures. PMID- 11940804 TI - Electroconvulsive Therapy and Prolactin Release: Relation to Treatment Response in Melancholia. AB - Serum prolactin levels 15 min after each of four consecutive electroconvulsive treatments (ECTs) in 16 male melancholics fell from the first to the fourth treatment. A larger mean prolactin response to ECT correlated with a slower treatment response, as measured by global outcome ratings after six ECTs and the total number of ECTs administered, both blind to prolactin levels. These findings were independent of baseline prolactin levels or treatment electrode placement; they are consistent with the hypothesis that ECT increases postsynaptic dopamine receptor sensitivity. Although intriguing, they await confirmation in a larger sample. PMID- 11940805 TI - Historical Article: Autobiography of L. J. Meduna. PMID- 11940806 TI - Potentiation of Seizure Length and Clinical Response to Electroconvulsive Therapy by Caffeine Pretreatment: A Case Report. AB - A patient with a history of nonresponse to electroconvulsive therapy (ECT) was treated with ECT modified by i.v. caffeine before electrical stimulation for some of the seizures. Seizures preceded by i.v. caffeine were longer, and led to marked clinical improvement. Caffeine pretreatment may be a method to enhance seizure length and efficacy in resistant patients. PMID- 11940808 TI - Clinical "Response" to "Convulsive" Therapy. PMID- 11940807 TI - Asthma/Emphysema Complication of Electroconvulsive Therapy: A Case Study. AB - The authors describe an elderly depressed patient with asthma/emphysema who developed severe postictal bronchospasm immediately after one electroconvulsive treatment. Possible problems and considerations in treating such patients are discussed. PMID- 11940809 TI - Computed Tomography and ECT. PMID- 11940810 TI - Reducing Memory Loss in Electroconvulsive Therapy. PMID- 11940811 TI - Time Course of Post-Electroconvulsive Therapy Prolactin Levels. AB - Serum samples before and after electroconvulsive therapy (ECT) from eight women patients revealed that prolactin blood levels reached a peak about 11 min after the end of the seizure. On average, the peak elevation over baseline was 385%; this elevation correlated strongly (r = 0.6, p < 0.001) with baseline blood levels. Prolactin peaks after bilateral ECT were significantly (p < 10(-5)) higher than those after unilateral ECT, although seizure duration was not longer. Application of a mathematical model gave an estimate of 17.3 min for the half life of prolactin elimination from the blood, close to published metabolic measurements. The rate of prolactin release into the blood was greater than the rate of its elimination from the blood. PMID- 11940812 TI - Electroconvulsive Therapy: Attitudes and Experience-A Survey of Indiana Psychiatrists. AB - The American Psychiatric Association survey concerning electroconvulsive therapy (ECT) was repeated with Indiana psychiatrists. The results indicated that these psychiatrists currently were more accepting of ECT than the earlier, national sample. However, there was agreement between the responses of the two groups on many of the issues. There were important differences between those who used ECT and those who did not. In Indiana, responders administering ECT were significantly more likely to be board certified than those who did not employ the technique. As expected, the users were, in general, more enthusiastic and favorable in their attitude toward ECT. The present survey suggests that there may be a greater awareness of adverse side effects and that improvements suggested in the literature are being adopted by practitioners. PMID- 11940813 TI - Electroconvulsive Therapy in Patients at High Risk for Physical Complications. AB - Nineteen patients had unrelieved incapacitating or life-threatening conditions likely remediable with electroconvulsive therapy (ECT). They also had coexisting medical conditions originally judged to preclude ECT because it risked unacceptable complications. ECT was administered with attempts to prevent complications, although some procedures were canceled for medical reasons. Fourteen of 19 patients completed their course of ECT; one patient died. Sixteen of 19 patients returned to baseline functioning and were discharged. Judicious attempts to treat such high-risk patients with ECT yield therapeutic gains. PMID- 11940815 TI - Anesthesia for ECT in General Hospitals. PMID- 11940816 TI - Multiple-Monitored ECT. PMID- 11940814 TI - Electroconvulsive Therapy and Prolactin Release: Effects of Stimulus Parameters. AB - We report a study of electroconvulsive therapy (ECT) induced prolactin release in nine patients as a function of treatment electrode placement, stimulus energy, and seizure length. Only electrode placement differentially affected prolactin release, with bilateral ECT yielding larger increments over baseline than unilateral ECT. PMID- 11940818 TI - Anesthesia in Electroconvulsive Therapy. PMID- 11940817 TI - Letters from the Orient. PMID- 11940819 TI - Electroconvulsive Shock and Brain Cholinergic Function: Role of Striatal Muscarinic Receptors. AB - Repeated administration of electroconvulsive shock (ECS) has been reported to reduce (3)H-quinuclidinyl benzilate ((3)H-QNB) binding to muscarinic cholinergic receptors in rat brain. ECS-induced muscarinic receptor subsensitivity may play a role in the therapeutic or adverse mechanisms of electroconvulsive therapy (ECT). In the present report, cataleptic responses to the muscarinic agonist, pilocarpine, were studied as a functional measure of brain muscarinic receptor sensitivity after ECS. Both single and repeated ECS administration attenuated pilocarpine-induced catalepsy, suggesting functional subsensitivity of the muscarinic receptors mediating this response. However, parallel changes were not observed in striatal muscarinic receptor binding. While repeated ECS induced a small but significant reduction in (3)H-ONB binding, single ECS marginally increased binding. The effect of pretreatment with a cholinergic antagonist (scopolamine) and with a cholinesterase inhibitor, diisopropyl fluorophosphate (DFP), on cataleptic responses was also studied. The findings demonstrated functional cholinergic subsensitivity after DFP administration and supersensitivity after scopolamine pretreatment. A clear relationship between functional alterations in muscarinic receptor sensitivity and radioligand binding parameters was not, however, established. Possible reasons for this discrepancy are discussed. PMID- 11940820 TI - Delusional Depression and Electroconvulsive Therapy: One Year Later. AB - This study reports the outcome of a 1-year follow-up study of 37 patients with a delusional depression treated with electroconvulsive therapy (ECT). Thirty-two (86%) of the patients initially responded to ECT. However, 16 (50%) of the 32 responders relapsed within 1 year. The authors suggest that further research is indicated to decrease this relapse rate. PMID- 11940821 TI - Relative Efficacy Rates for Therapies of Delusional Depression. AB - The treatment responses of 597 patients with delusional depression from 17 studies were compared. Patients were treated with either tricyclic anti depressants (TCAs), antipsychotics (APs), the combination of TCA/AP, or electroconvulsive therapy (ECT). The overall responses were 34% in the group treated with TCA, 51% in the AP group, 77% in the TCA/AP group, and 82% in the ECT group. Prospective studies to define the risk/benefit ratios of the more effective therapies, TCA/AP and ECT, are needed. PMID- 11940822 TI - Early Change in Cognitive Performance Accompanying Bilateral ECT. AB - In order to characterize the onset and evolution of early cognitive changes that accompany bilateral electroconvulsive therapy (ECT), the performance of 13 ECT treated and 16 psychiatric control patients on a battery of neuropsychological tests administered daily was compared. These tests included: Digit Symbol, spokes, letter cancellation, and short-term memory tests. While both groups improved on all tests, performance of ECT patients vs. controls on letter cancellation tests diverged significantly (p < 0.005) after three ECTs. The authors speculate that this finding reflects an alteration in attentional processes caused by bilateral ECT. PMID- 11940823 TI - Can Patients Safely Undergo Electroconvulsive Therapy While Receiving Monoamine Oxidase Inhibitors? AB - A review of the experience of monoamine oxidase inhibitors (MAOI) and anesthesia finds few well-documented complications, although theoretic risks of cardiovascular reactions are discussed. Laboratory studies and actual case experiences of patients treated with electroconvulsive therapy (ECT) and MAOI suggest that the risks have been exaggerated. It is recommended that the anesthesia usually used for ECT-methohexital, succinylcholine, and optionally atropine or glycopyrrolate-may be safely given with careful blood pressure monitoring. If cardiovascular support is required, a direct-acting sympathomimetic should be administered. PMID- 11940824 TI - Psychiatric Malpractice: The Electroconvulsive Therapy Experience. AB - The malpractice experience of the American Psychiatric Association professional insurance program from 1972 to 1983 is reviewed. Seventeen cases among 711 "closed" cases related to electroconvulsive therapy (ECT). Nine were settled in favor of the psychiatrist; one case was tried and the jury found for the psychiatrist. Patients prevailed in seven cases by settlement. The author concludes that malpractice losses are not a significant factor in ECT practice or in its negative public image. PMID- 11940825 TI - Convulsive Therapy: Fifty Years of Progress. AB - The origins of convulsive therapy, its early developments, present practice, central elements of the treatment, and neurohumoral and neuroendocrine theories of its antidepressant action are reviewed on the 50th anniversary of its introduction. PMID- 11940827 TI - National Institutes of Health Consensus Conference. PMID- 11940826 TI - Electroconvulsive Therapy with Fetal Monitoring in a Bipolar Pregnant Patient. AB - The management of depression in a pregnant patient is a therapeutic challenge because of the potential teratogenicity of psychotropic medications. The present case report details the safe and effective use of electroconvulsive therapy in a pregnant patient with bipolar affective disorder. Values for maternal blood gases, fetal heart rate monitoring with Doppler ultrasound, and the use of real time ultrasonography indicate that the treatment has no adverse effect on the fetus. This is believed to be the second case report in the literature in which continuous fetal monitoring and maternal blood gases have been used during electroconvulsive therapy. PMID- 11940828 TI - Electroconvulsive Therapy and Electric Acupuncture Convulsive Therapy in China. AB - This essay presents briefly the historical development and the present condition of electroconvulsive therapy (ECT) and Electric Acupuncture Convulsive Therapy (EACT) in China. Written on the basis of internal published documents and materials concerning ECT and EACT since 1950 and the clinical experience and experimental research of the authors, it puts forward the authors' views of problems that remain in applying ECT. PMID- 11940829 TI - Electric Acupuncture Convulsive Therapy. AB - In 150 schizophrenic patients, a comparative investigation between electric acupuncture convulsive therapy (EACT) and electroconvulsive therapy (ECT) showed that the current used for eliciting a convulsion in EACT was only 3.6% of that for ECT when the electrodes were placed at acupoints Baihui and Renzhong. EACT is a modification of ECT in which stimulating currents are passed through acupuncture needle electrodes inserted in midline positions. In this study, the efficacy of EACT was better, the somatic and visceral reactions milder, and the incidence of spine fracture and changes in EEG and in memory were less than in ECT. The clinical efficacy of electroconvulsive therapy is seen to depend on changes in midline brain structures. PMID- 11940830 TI - Characterization of the Total Amount of Prolactin Released by Electroconvulsive Therapy. AB - Many factors can obscure determination of the total hormone release induced by a course of electroconvulsive therapy (ECT). Not the least of these factors are the rates of hormone release and their elimination. A method of estimating the total amount of prolactin released during an ECT-induced seizure has been constructed by mathematical modeling. Using prolactin levels measured from patients undergoing ECT, this simulation indicates that the optimal times for sampling plasma prolactin are 5, 10, 20, and 45 min following the seizure, besides the baseline measurement. PMID- 11940831 TI - Electroconvulsive Therapy During Anticoagulant Therapy. AB - The safety of ECT in patients receiving anticoagulants is reviewed. In patients who failed to respond to drug therapy and in some whose cardiovascular conditions precluded the use of antidepressant drugs, the authors successfully carried out five courses of ECT in depressed patients maintained on anticoagulants. In three cases, heparin was used, blood coagulation values were closely monitored, and the dose of succinylcholine employed was increased to provide almost complete muscular paralysis during the seizure. In one patient, acenocoumarol was sustained. The authors conclude that ECT may be safely administered in the presence of anticoagulant drug therapy. PMID- 11940832 TI - Efficacy of Electroconvulsive Therapy in Schizophrenia, Mania, and Other Disorders. I. Schizophrenia. AB - Clinical studies are reviewed comparing the efficacy of ECT and sham ECT, ECT and neuroleptic drugs, and the effects of electrode placement in the therapy of patients with schizophrenia. The efficacy of ECT is greater in acutely ill patients, and combined with neuroleptics it elicits a better short-term outcome than drug treatment alone. No differential effects of electrode placement are noted. The need for more sophisticated evaluations is emphasized. PMID- 11940833 TI - Efficacy of Electroconvulsive Therapy in Schizophrenia, Mania, and Other Disorders. II. Mania and Other Disorders. AB - Clinical studies of the efficacy of ECT in mania are reviewed. ECT is effective in the short-term treatment of mania, but its use may require concurrent neuroleptic drugs early in treatment as the effects of both ECT and lithium are delayed. Indications for ECT in delirium, post-partum illnesses, psychoneuroses, and other conditions are also reviewed. PMID- 11940834 TI - Nursing Students' Perception of Electroconvulsive Therapy: Impact of Instruction with an Electroconvulsive Therapy Videotape. AB - There has been little study of attitudes toward and knowledge of electroconvulsive therapy (ECT). Likewise, there have been no studies of how accurate information about ECT may be communicated. The authors studied the attitudes and opinions toward ECT of thirty-five nursing students before they had received any formal instruction on its use. The same subjects then viewed a didactic videotape explaining and demonstrating ECT. There were significant improvements in overall knowledge and positive opinions of ECT after this educational exposure. PMID- 11940835 TI - ECT Risks in Glaucoma. PMID- 11940836 TI - Multiple-Monitored ECT. PMID- 11940837 TI - ECT in Italy. PMID- 11940838 TI - Convulsive Therapy in China: What are the Implications? PMID- 11940839 TI - Does Electroconvulsive Therapy Prevent Suicide? AB - To examine the issue of whether or not electroconvulsive therapy (ECT) protects against suicidal death, we followed a complete population of 1,494 adult hospitalized psychiatric patients for 5-7 years. During that time there were 76 deaths, of which 16 or 21% were by suicide. Cause of death was not significantly related to age, gender, or research diagnosis. Patients who committed suicide were more apt to have received ECT than those who died from other causes, but this difference was not significant. A control group of living patients matched for age, sex, and diagnosis had very similar exposures to ECT, which further indicates that ECT does not influence long-term survival. These findings combined with a close examination of the literature do not support the commonly held belief that ECT exerts long-range protective effects against suicide. PMID- 11940840 TI - Meta-Analytical Approach to Reconciling Discrepancies in Efficacy between Bilateral and Unilateral Electroconvulsive Therapy. AB - A review of the literature assessing the efficacy of bilateral compared to nondominant unilateral ECT was conducted using a quantitative review procedure meta-analysis. The results supported conventional reviews that concluded that there is no difference in outcome between bilateral and unilateral ECT. Meta analysis also allowed an evaluation of the effects of 11 variables that span: (a) research methodology, (b) technique, (c) patient demographics, and (d) operational dimensions that have been proposed to mediate results of studies that found an advantage for bilateral over unilateral ECT. Of these variables, shorter interelectrode distance and the assessment of outcome after a fixed number of treatments (e.g., five or six ECT) rather than after completion of the full ECT course were significantly related to studies reporting a clinical advantage for bilateral ECT, and together, both accounted for 45% of the variance across studies. Although weaker effects of unilateral ECT due to shorter interelectrode distances may be related to seizure threshold, assessment after a fixed number of treatments may be related to failure to monitor seizure length. PMID- 11940841 TI - Effect of Training on Administration of Electroconvulsive Therapy by Junior Doctors. AB - An instrument was designed to rate the administration of electroconvulsive therapy. The performance of junior doctors was rated before and after videotape training in technique. The group showed a marked increase in ratings following training. The limitations and importance of the study are discussed. PMID- 11940842 TI - Informing Patients About Electroconvulsive Therapy: Effects of a Videotape Presentation. AB - Issues of information given for consent purposes are particularly problematic with electroconvulsive therapy (ECT). The authors investigated the efficacy of an informative videotape recording in this process. The videotape contained factual information about ECT, an interview with a patient before and after treatment, and the presentation of an actual treatment. A patient group assigned to watch the videotape, in addition to the usual consent process involving written and verbal information from the treating resident psychiatrist, showed only one postconsent difference from a group that had the usual consent process without the videotape. Those who saw the videotape were less sure that they had adequate information to decide whether to have ECT than those who did not see it (p < 0.05). Although patients likely to benefit from such an audiovisual approach may be identifiable, it seems that an informative videotape offers no universal advantage over more conventional information giving in the consent process. PMID- 11940843 TI - Attitude Studies on Electroconvulsive Therapy. AB - Between 1971 and 1984, 11 studies have examined patient and/or lay attitudes to electroconvulsive therapy. These investigations are reviewed here. They vary in sample size, response rates, and in the way attitudes were assessed. Although none have used properly validated measures, the uniformity of their findings provides a considerable degree of validation. Studies have been conducted in the United States, United Kingdom, Eire, Australia, and Uganda. In general, patients felt positively about the treatment, and for most subjects the benefit of the treatment outweighed the cost, in terms of apprehension, side effects, and stigma. For a small, but important, minority, this was not so, and such individuals report quite negative attitudes to ECT. All the studies reviewed have faults, and suggestions for further research are outlined. PMID- 11940844 TI - Initial Impression of Two New Brief-Pulse Electroconvulsive Therapy Machines. AB - Until last year only two U.S. manufacturers were producing ECT machines for purchase. These machines differed in waveforms: a sine-wave device (Medcraft Corp.) and a brief-pulse bidirectional square-wave machine (MECTA Corp.). Early in 1985 a new brief-pulse bidirectional square-wave machine called the Thymatron was marketed (Somatics Inc.). For the first time this created a choice in brief pulse equipment available in the U.S. This paper summarizes our initial impressions to this new brief-pulse device and one of MECTA's most recent brief pulse models (SR-1) in terms of features, safety, practicality, and cost. PMID- 11940845 TI - Double Electroconvulsive Therapy for Resistant Depression. AB - A case is described of psychotic depression that worsened during a course of three bilateral ECTs per week, but responded to administration of two bilateral ECTs three times per week. For some patients, ECTs may need to be given more than three times per week to obtain remission. PMID- 11940846 TI - ECT and Suicide. PMID- 11940847 TI - Electroconvulsive Therapy in Treating Neuroleptic Malignant Syndrome. PMID- 11940849 TI - A New Evaluation of Convulsive Therapy: The Ontario Report. PMID- 11940848 TI - Electroconvulsive Therapy in Great Britain, 1981 Follow-Up. PMID- 11940850 TI - Quantitative Neuropathology in Electrically Induced Generalized Convulsions. AB - A critical review of the neuropathology associated with seizures is presented, with special reference to the damage to the brain claimed to result from electroconvulsive treatment (ECT). From the point of view of neuron loss, there is no evidence that single seizures (convulsions), even if repeated, result in this type of damage to the brain, which is the only definitely irreversible kind. Pitfalls of the methods of investigation available in neuropathology are described to support this statement. Basic differences between epileptic seizures and induced seizures are emphasized in relation to neuron loss in epilepsy. PMID- 11940851 TI - Electroconvulsive Therapy Effects on Cerebrospinal Fluid Monoamine Metabolites and Platelet Serotonin Uptake In Melancholia. AB - The effects of ECT on 5-hydroxyindoleacetic acid, homovanillic acid (HVA), and 4 hydroxy-3-methoxyphenylglycol in cerebrospinal fluid and on kinetic parameters of platelet serotonin uptake were studied in 12 patients with melancholia. There were no significant changes in the monoamine metabolites 3 weeks after initiation of ECT in 12 patients; however, there was a tendency for HVA to increase. The V(max) of serotonin uptake (measured in seven patients) remained unchanged after ECT, but there was a significant increase in K(m), indicating a decreased affinity for serotonin in the carrier. PMID- 11940852 TI - Electrode Placement and Prolactin Response to Electroconvulsive Therapy. AB - Study of serum prolactin during electroconvulsive therapy (ECT) in depressive patients revealed a greater prolactin increase after bilateral than after unilateral ECT. A linear correlation between the two types of prolactin response was found for a group of 10 patients, a finding that suggests a quantitative rather than a qualitative difference between bilateral and unilateral ECT with regard to their prolactin-releasing properties. The magnitude of prolactin response did not differ between right and left unilateral ECT, nor in a systematically studied case of postictal dysphoric excitement that occurred after right, but not after left, unilateral ECT. In this case, maximal prolactin response occurred earlier with right than with left unilateral ECT. Prolactin increase after ECT was not correlated with such factors as severity of depression nor seizure duration. PMID- 11940854 TI - The Electroconvulsive Therapy Review Committee in Ontario, Canada. PMID- 11940855 TI - An Auditory Representation of ECT-Induced Seizures. AB - Seizure monitoring using an auditory representation of the electroencephalogram (EEG) was tested in 82 consecutive electroconvulsive treatments (ECTs). It was found to have high interrater and intrarater reliability and high validity against the paper EEG standard. PMID- 11940853 TI - ECT: Clinical Variables, Seizure Duration, and Outcome. AB - The ability to predict treatment response in patients administered electroconvulsive therapy remains limited. Variables such as the duration of the individual seizure or the cumulative seizure duration have been reported to relate to outcome in a "threshold" or "therapeutic window" model. In this study, total seizure time, as measured by bifrontal electroencephalogram (EEG) electrodes, did correlate with outcome (r(s) = 0.43, p < 0.01). In addition, 88% of patients with a cumulative seizure time exceeding 300 s responded favorably (chi(2) = 6.80, p < 0.01). This provides support for the concept of a minimum threshold of accumulated seizure time necessary for clinical response. However, no evidence of an upper limit of benefit suggesting a "therapeutic window" was found. PMID- 11940856 TI - Electrotherapy Certification. PMID- 11940857 TI - ECT and Glaucoma. PMID- 11940858 TI - Differential Ability of ECT Devices to Produce Seizures. PMID- 11940859 TI - Optimum Frequency of Electroconvulsive Therapy: Implications for Practice and Research. PMID- 11940860 TI - EEG-Monitored ECT: A Comparison of Seizure Duration under Anesthesia with Etomidate and Thiopentone. AB - Electroencephalogram-monitored electroconvulsive therapy (ECT) was carried out in 20 depressed inpatients. Before treatment, patients were randomly allocated to treatment using etomidate (Hypnomidat) (n = 10) or thiopentone (n = 10) for anesthesia. The groups were matched for sex, age, weight, and type and severity of depression. The seizure duration (seconds) was measured by electroencephalography (EEG), and the electrical energy (Joules, J) was determined for each treatment. A ratio of seizure duration:electrical energy (s/J) was computed. Both seizure duration and seizure duration:electrical energy were greater in the etomidate group than in the thiopentone group, whereas electrical energy did not differ significantly. The number of treatments in the etomidate group did not differ from that in the thiopentone group, as may be expected, perhaps because of the small size. PMID- 11940861 TI - Dexamethasone Suppression Test as a Predictor of Response to Electroconvulsive Therapy. I. Inpatient Treatment. AB - Thirty-eight psychiatric inpatients who fulfilled DSM-III criteria for major depressive disorder with melancholia received weekly dexamethasone suppression tests (DSTs) while undergoing a clinical course of ECT. Blind ratings of clinical improvement were obtained weekly from patients on the Beck Depression Inventory and from physicians and nurses on the Hamilton Depression Scale. The 26 patients with abnormal DSTs and the 12 patients with normal DSTs demonstrated an equivalent rate of clinical remission (75 vs. 77%) as defined by a composite rating of the nurse, physician, and patient. Other individual rater analyses also confirmed the finding that initial DST status is not predictive of response to a course of ECT. PMID- 11940863 TI - Electroconvulsive Therapy in a Sample British Population in 1982. AB - This study into the practice of electroconvulsive therapy (ECT) within a sample British population of three million found that although the use of ECT had declined by half in the past decade, it was still prescribed frequently. Closer scrutiny of a subsample of 390,000 found it to be applied mainly to the treatment of major depressive illness in elderly women. The referring clinicians demonstrated a preference for bilateral electrode placement, with treatment being administered thrice weekly. It was common practice to prescribe concurrent psychoactive medication. The clinicians predicted favourable initial outcomes with 90% accuracy whatever the presenting diagnosis or symptoms. High overall mortality and morbidity of the depressed patients was noted. Almost all (93%) of the treatment courses were administered to informal patients. PMID- 11940862 TI - Dexamethasone Suppression Test as a Predictor of Response to Electroconvulsive Therapy. II. Six-Month Follow-Up. AB - In an earlier study, the dexamethasone suppression test, obtained 3-6 days after hospital admission, did not reliably predict the proportion of clinical remissions or the degree of improvement experienced by severely-depressed inpatients undergoing a course of ECT. In this regard, about 75% of initial suppressors and nonsuppressors showed remission, and those nonsuppressors who subsequently normalized did not show a better clinical response than persistent nonsuppressors. In keeping with the findings of Coryell and Zimmerman (1983) we also found that persistent nonsuppressors had a better clinical outcome 6 months after discharge than did initial nonsuppressors who normalized by discharge. PMID- 11940864 TI - The Experience of Electroconvulsive Therapy in the 1980s: A Prospective Study of the Knowledge, Opinions, and Experience of California Electroconvulsive Therapy Patients in the Berkeley Years. AB - In 1982, the city of Berkeley, California voted to make the use of electroconvulsive therapy (ECT) a crime. Though later overturned, this ordinance generated much publicity and underscored the public's general negative view of ECT. In this climate, the authors surveyed first-time ECT recipients (n = 35), and a group with prior ECT (n = 20) to examine patient knowledge, experience, and opinions of ECT in California in the 1980s. Patients in both groups believed ECT should be available and did not think it was used for punishment or control. First-time ECT patients showed good understanding of the ECT process, thought that they had adequate information with which to decide about ECT, and were optimistic about the outcome. In comparison, those with prior ECT were less knowledgeable, less sure about the ECT process, less optimistic about the outcome, and more frightened of the procedure. After treatment, first-time ECT patients believed they had made a good decision to have ECT and that they were helped. Complaints of memory dysfunction were common. Patients who received ECT in the past may have had a more negative experience with ECT than those undergoing the procedure in the 1980s. These more negative experiences may have determined present media presentations and, therefore, had an influence on present day public attitudes against ECT. Accurate portrayals of ECT, as it is done today, may improve public attitudes toward ECT and forestall adverse legislation. PMID- 11940865 TI - Differences Between First and Second Electroconvulsive Treatments Given in the Same Session. AB - For the first and second of two bilateral ECTs given under the same anesthesia, judgments were made of the durations of four seizure-induced phenomena: tonic clonic motor activity, paroxysmal EEG activity, EEG spike waves, and tachycardia. A total of 36 pairs of treatments were given to 14 patients. Pearson correlation coefficients between each of six possible pairs of these four measures were higher for the first ECTs than for the second ECTs (p < 0.05 to p < 0.001). These results suggest that the seizure associated with the second ECT under the same anesthesia is physiologically different from that of the first ECT and is less well generalized through the brain. PMID- 11940866 TI - Electroconvulsive Therapy and Cerebral Venous Angioma. AB - A 24-year-old man with a left parietal cerebral venous angioma underwent a course of electroconvulsive therapy (ECT) for the treatment of bipolar affective disorder. We were able to treat this patient safely and successfully with ECT. There was no evidence of a complication related to his vascular malformation, as was demonstrated by follow-up MRI examination. Our experience suggests that ECT may be safely instituted in patients with venous angiomas, and changes in their lesions can be safely examined by MRI. PMID- 11940867 TI - Prolonged T-Wave Inversion Associated with Electroconvulsive Therapy. AB - A patient is presented who developed diffuse T-wave inversion during a course of ECT. Review of the literature suggests that prolonged T-wave inversion associated with ECT is an uncommon and probably benign finding not warranting an interruption of a course of electrotherapy. PMID- 11940868 TI - Electroconvulsive Therapy in Osteogenesis Imperfecta: Issues of Muscular Relaxation. AB - The authors describe a depressed patient with osteogenesis imperfecta, osteoporosis, and vertebral compression fractures, who was successfully treated with electroconvulsive therapy (ECT). The management during ECT of patients at increased risk of fractures is discussed. PMID- 11940869 TI - Training in Convulsive Therapy. PMID- 11940870 TI - ECT in Unipolar and Bipolar Disorders: A Naturalistic Evaluation of 460 Patients. AB - We compared the efficacy of electroconvulsive therapy (ECT) and numbers of ECTs received by unipolar depressive (n = 368), bipolar depressive (n = 55), and manic patients (n = 37) in a review of records of patients treated in a 12-year period in a university hospital medical center. Both unipolar and bipolar depressive patients received nine treatments, one half more on average than the number of treatments received by manic patients, but the difference was not significant. ECT was equally effective in unipolar and bipolar depression (69.8 and 69.1%, respectively, rated as "markedly improved"), whereas 78.4% of patients with mania had "marked improvement." Both unilateral and bilateral ECTs were equally effective for the three groups. Bipolar depressive and manic patients receiving mixed courses (some unilateral, some bilateral) received more treatments than did those receiving unilateral or bilateral treatments exclusively and tended not to respond as well. We conclude that ECT is an effective treatment for mania, unipolar, and bipolar depression, that unilateral and bilateral treatments are equally effective, and that no significant difference exists in the number of ECTs used to treat these disorders. PMID- 11940871 TI - Practice Patterns of Electroconvulsive Therapy: A California Perspective (1984). AB - The sixty-five California hospitals administering electroconvulsive therapy (ECT) during 1984 were surveyed regarding ECT practices to develop data on the community standard of care. Usable data were obtained from 58 hospitals. In each facility two psychiatrists usually administered ECT, including one psychiatrist who received credentials in or after 1977. Thirty-four brief-pulse and 43 sine wave machines were in use. Treatment was performed on the psychiatric unit in 41.4% of the hospitals, in the operating room or the recovery room in 50% of the hospitals, and in other locations in 8.6% of the hospitals. The psychiatrist administered the anesthesia in 12 (21%) of the hospitals. Unilateral electrode placement was used in 42% of patients. The use of anesthetic agents, ancillary equipment, and staffing patterns is reviewed. PMID- 11940872 TI - A Comment on the Efficacy of Unilateral Versus Bilateral ECT. AB - Ten studies comparing the efficacy of unilateral versus bilateral electroconvulsive therapy (ECT) were reexamined. Three different methods of meta analysis applied to the combined results revealed statistical significance in favor of bilateral ECT for the relief of depression. The recognition that bilateral ECT has some advantage over unilateral is in sharp contrast to an overly strong conclusion to the contrary previously reached by Janicak et al. (1985), in reviewing the same studies. Alternative statistical methods for evaluating the significance of combined results from several independent studies are illustrated with reference to previously published ECT research. PMID- 11940873 TI - A Hypothesis to Explain Divergent Findings Among Studies Comparing the Efficacy of Unilateral and Bilateral ECT in Depression. AB - This article explains divergent reports on the relative efficacy of unilateral and bilateral electroconvulsive therapy (ECT) by hypothesizing a therapeutic effect of both seizure and electrical stimulus, with the seizure's effect based on its intensity and generalization. The effect of the stimulus is normally obscured by the seizure but emerges with reduced electrical dosage or elevated threshold. These circumstances favor a therapeutic advantage for bilateral ECT, which introduces more electrical charge into a larger and differently distributed volume of brain and induces greater seizure generalization, more diencephalic stimulation, and a relative decrease in left relative to right hemisphere electroencephalographic (EEG) frequencies. PMID- 11940874 TI - Commentary on a Hypothesis to Explain Divergent Findings Among Studies Comparing the Efficacy of Unilateral and Bilateral ECT in Depression by Richard Abrams, M.D. PMID- 11940875 TI - Minimizing Therapeutic Differences Between Bilateral and Unilateral Nondominant ECT. AB - Conflicting results continue to be reported for studies contrasting the therapeutic efficacy of bilateral and unilateral nondominant electroconvulsive therapy (ECT). At least in part, the therapeutic advantage for bilateral ECT observed by some investigators may be related to the use of nonoptimum unilateral ECT technique. Consideration of technique-related factors, such as stimulus electrode location, contact at the electrode-scalp interface, stimulus dosing, and seizure monitoring, will allow unilateral ECT to be carried out with maximal therapeutic potency. PMID- 11940876 TI - Neurophysiological Variability in the Effects of the ECT Stimulus. AB - There is evidence that, in addition to seizure elicitation, critical factors in therapeutic response to ECT may include aspects of stimulus parameters and electrode placement. Dr. Abrams suggests that these factors contribute to efficacy by influencing the degree of seizure generalization, diencephalic stimulation, and relative asymmetry in the slowing of EEG frequencies. An alternative view is presented here, which hypothesizes that these factors converge in influencing the regional distribution and strength of the endogenous inhibitory process that terminates the seizure. PMID- 11940877 TI - Positive Response to Bilateral Sinusoidal ECT in Unilateral and Bilateral Brief pulse "ECT-Resistant" Depressive Illness. AB - Six patients who failed to respond to what would ordinarily be considered therapeutically effective courses of sequential unilateral nondominant (mean number 10) and bilateral brief-pulse electroconvulsive therapy (ECT) (mean number 10) with electroencephalographic (EEG) monitoring of seizure duration (all seizures >30 s) subsequently received courses of bilateral sinusoidal ECT (mean number 6) and responded well. Some theoretical implications and clinical considerations raised by these cases are discussed. PMID- 11940878 TI - Electroconvulsive Therapy and Severe Osteoporosis: Use of a Nerve Stimulator to Assess Paralysis. AB - An elderly patient at high risk for fractures due to severe osteoporosis and other spinal pathology was treated with electroconvulsive therapy (ECT). Suppression of twitch response to peripheral nerve stimulation over the ulnar nerve was a more accurate monitor of degree of paralysis than other clinical indices. Complete paralysis was consistently achieved using this method. Successive doses of succinylcholine were not required with multiple treatments per session when the nerve stimulator was used to verify continued paralysis. PMID- 11940880 TI - A Survey of Electroconvulsive Therapy: Use and Training in University Hospitals in 1984. PMID- 11940879 TI - ECT-Induced Myoclonus. AB - We report a case of electroconvulsive therapy (ECT)-induced myoclonus which occurred postictally following every treatment and was distinguished from continuing seizure activity by electroencephalogram (EEG) recordings. Subsequent treatment with amitriptyline resulted in an isolated episode of myoclonus that did not recur when the dosage of medication was increased. Besides the clinical implications, this sequence of events raises questions about the serotonin hypothesis of antidepressant-induced myoclonus. PMID- 11940881 TI - Pheochromocytoma, Depression, and Electroconvulsive Therapy. PMID- 11940882 TI - Transference Reactions to an ECT Apparatus. PMID- 11940883 TI - Uni- Versus Bilateral ECT in the Treatment of Mania. AB - Recent reports have challenged the assumption that unilateral and bilateral electroconvulsive therapy (ECT) are equally effective in all cases. In a prospective study, hospitalized manic patients were randomly assigned to treatment with either lithium or ECT. The first six patients received nondominant unilateral treatment but failed to respond. However, they rapidly remitted when switched to bilateral ECT. Accordingly, the next 11 manic patients to receive ECT were assigned to bilateral treatment. They were compared with the first six patients on weekly ratings of mania and other psychopathology. Both ECT groups improved but therapeutic responses were delayed until after switchover to bilateral treatment in the patients who began with unilateral ECT. We conclude that patients in the manic state should receive bilateral rather than unilateral ECT. PMID- 11940884 TI - Atropine Sulfate Premedication and Cardiac Arrhythmia in Electroconvulsive Therapy (ECT). AB - Prophylactic effects of atropine sulfate as a premedication for cardiac arrhythmia were studied in 44 patients receiving electroconvulsive therapy in a randomized, cross-over, double-blind design. Cardiac rhythms were recorded with Holter monitors during atropine sulfate and placebo treatments. Vagal arrhythmia seemed to be reduced with atropine sulfate premedication, but the frequency of ventricular arrhythmia was not affected. PMID- 11940885 TI - Reliability and Validity of Measures of Seizure Duration. AB - Two raters independently assessed seizure duration in single-channel electroencephalogram (EEG) records obtained in 479 electroconvulsive treatments of 52 patients. Ratings of seizure duration were highly reliable. The measures of EEG seizure duration were substantially associated with the duration of motor manifestations, supporting the validity of the EEG determinations. Confidence ratings of the clarity of the endpoint of EEG paroxysmal activity were unrelated to the duration of seizures in EEG and motor measures. The extent to which the EEG seizure activity extended beyond the motor manifestations was strongly related to the total duration of the EEG seizure manifestations and not to the duration of motor manifestations. PMID- 11940886 TI - Increased Incidence of Missed Seizures During ECT in the Elderly Male. AB - Monitoring seizure length is an important addition to the clinical practice of ECT. Without monitoring, undetected missed seizures may occur. In a retrospective study of ECT treatments for a sample of 58 patients with a DSM-III diagnosis of major depression, significantly more missed seizures (p < 0.05) occurred in the elderly males. PMID- 11940887 TI - Electroconvulsive Therapy in Nigeria. AB - An overview of the use of electroconvulsive therapy (ECT) in Nigeria is presented. Unmodified ECT with bilaterally placed electrodes remains the main method of administration and is widely used for all forms of functional psychoses. ECT is used as an adjunct to chemotherapy both for inpatients and outpatients. Complications associated with the therapy are relatively few. Despite the use of unmodified ECT since 1952, there has been no report of tardive (spontaneous) seizures in patients treated by this method. In view of the widespread usage, a continuing assessment of the therapy and guidelines for its use in Nigeria seem desirable. PMID- 11940888 TI - Combined Lithium and Electroconvulsive Therapy: Pharmacokinetic and Pharmacodynamic Interactions. AB - The combined use of lithium and electroconvulsive therapy (ECT) is currently contraindicated, due to reports of associated neurotoxicity and poor outcome. Only limited data are available to explain any pharmacological basis for an adverse interaction between the two treatments. ECT does not alter lithium distribution or kinetics. Possible drug-drug interactions between lithium and ECT premedication have been subjected to little systematic study. Despite a body of preclinical work implicating the cholinergic system in the mechanism of action of both lithium and ECT, no interaction between lithium and pre-ECT anticholinergic medication has been reported. Potentiation of barbiturate anesthetics and neuromuscular blocking agents, including succinylcholine, by lithium has been noted in the anesthesiology literature, but reports of such interaction are lacking in the context of ECT administration. Thus, no demonstrable pharmacokinetic or drug-drug interaction factors preclude combined prescription of lithium and ECT. PMID- 11940889 TI - Electroconvulsive Therapy in Two Former Neurosurgical Patients: Skull Prosthesis and Ventricular Shunt. AB - Electroconvulsive therapy (ECT) was successfully used to treat patients with prior neurosurgical history. The first patient had undergone surgery for a meningioma with subsequent insertion of a plate in the cranium. The second patient had a history of normal pressure hydrocephalus which was treated with a ventriculoperitoneal shunt procedure. Both patients received ECT and in both cases the presenting symptoms of depression abated. Treatment was considered both uneventful and successful. PMID- 11940890 TI - Atracurium as a Muscle Relaxant for Electroconvulsive Therapy in a Burned Patient. AB - We present the case of a patient with 80% total body surface area burned who presented for anesthesia and electroconvulsive therapy (ECT) 75 days after the injury. He was presumed to be sensitive to the K(+)-releasing effects of succinylcholine and therefore was given a nondepolarizing neuromuscular blocking agent, atracurium, to which he was shown to be resistant. Anesthesia for ECT on 13 occasions was without incident. Sensitivity and K(+) release secondary to succinylcholine is discussed as well as resistance to non-depolarizing neuromuscular blocking agents. PMID- 11940891 TI - ECT Modified by Atracurium. AB - Present administration of electroconvulsive therapy (ECT) is safe for most patients. Succinylcholine is used and tolerated as one of several pharmacologic agents in the routine protocol. Relatively rare medical conditions, including pseudocholinesterase deficiency, cholinesterase inhibition, and severe neuromuscular disease, may complicate the use of succinylcholine. This article presents three cases in which the standard protocol was altered, using atracurium in place of succinylcholine to provide brief and well-controlled muscle relaxation. Atracurium is an alternative medication for ECT in those cases in which succinylcholine use is problematic. PMID- 11940892 TI - Use of ECT to Treat Bipolar Depression in a Mental Retardate with Cerebral Palsy. AB - We report the use of electroconvulsive therapy (ECT) for treatment of a bipolar episode in a mildly retarded young man with cerebral palsy. Difficulties in the diagnosis and treatment of affective disorders in the retarded are discussed. PMID- 11940894 TI - Meta-Analysis of Effects of Electrode Placements. PMID- 11940893 TI - Postictal Delirium (and Recovery From Melancholia) After Left-Unilateral ECT. AB - Postictal delirium was observed after left-unilateral ECT in a fully dextral man whose melancholia entirely remitted with seven treatments. These findings suggest that it is premature to attribute either the syndrome of postictal delirium or the beneficial effects of ECT in melancholia primarily to right hemisphere mechanisms. PMID- 11940895 TI - A Reply to Mathisen and Pettinati. PMID- 11940896 TI - Multiple-Monitored ECT. PMID- 11940897 TI - A Reply to Maletzky. PMID- 11940898 TI - Optimum Frequency of Electroconvulsive Therapy. PMID- 11940900 TI - Neuropathology of ECT. PMID- 11940899 TI - A Reply to Stromgren. PMID- 11940902 TI - Treatment Options in Severe Psychotic Depression. AB - In our sample of 15 hospitalized patients with severe psychotic depression, six responded to therapy with tricyclic antidepressants combined with neuroleptics. Of the nine nonresponders, eight showed an excellent clinical response to electroconvulsive therapy (ECT). The clinical outcome after 6 months of treatment was similar in both groups. We recommend that protracted, complicated trials of pharmacotherapy be reevaluated in psychotic depression. A prospective comparative study of ECT and pharmacotherapy is needed to define the optimal treatment for psychotic depression. PMID- 11940901 TI - Neuropsychiatry and Behavioral Neurology. PMID- 11940903 TI - Subjective Side Effects Acutely Following ECT: Associations with Treatment Modality and Clinical Response. AB - Patient reports of systemic side effects and cognitive impairment were obtained the afternoon of each treatment during the electroconvulsive therapy (ECT) course. Side effects were grouped a priori as reflecting physical complaints, perceived cognitive impairment, and as mood-related. Patients randomly assigned to bilateral ECT reported more cognitive impairment than patients who received right unilateral ECT. There were indications that the unilateral ECT group had greater physical/somatic complaints. Patients who responded to ECT differed from nonresponders only on the more explicitly mood-related items. Generally, the findings supported the desirability of extending studies of patient reports of ECT adverse effects to include subjective systemic side effects in addition to subjective cognitive impairment. PMID- 11940904 TI - Serial Dexamethasone Suppression Tests and Clinical Outcome in ECT. AB - Serial dexamethasone suppression tests (DST), obtained during a course of electroconvulsive therapy in 43 severely depressed patients, did not exhibit relationships between the initial DST, final DST, or the change in DST with clinical outcome measures at the time of discharge. In 37 patients reviewed six months after discharge, no relationship with the continuation of improvement, rehospitalization, or suicide was found. We are unable to confirm a clinical application for the DST in the management of patients during a course of convulsive therapy. PMID- 11940905 TI - ECT and Neurological Disorders. AB - Clinicians become concerned when ECT is contemplated in an individual with a neurological disorder. In this review, the authors summarize the reports on the use of ECT in the presence of neurological disease. Because blood pressure, cerebral blood flow, and intracranial pressure rise with ECT, space-occupying lesions with increased intracranial pressure, cerebral aneurysm, recent head trauma, or active CNS infection pose special concerns for ECT treatment. In this review, we conclude that epilepsy and states with lowered seizure threshold may predispose to prolonged seizures. A history of head injury or stroke probably does not increase risk. Toxic/metabolic disorders are not contraindications to ECT, although correction of the underlying imbalance is a first priority. Extrapyramidal, demyelinating, and neuromuscular disorders pose little increased risk. Indeed, in Parkinson's Disease, ECT may be beneficial for the motor symptoms. As a general rule, it seems advisable to treat the underlying disorder prior to beginning ECT. PMID- 11940906 TI - Electroconvulsive Therapy in Multiple Sclerosis: A Magnetic Resonance Imaging Study of the Brain. AB - A depressed woman with multiple sclerosis was successfully treated with electroconvulsive therapy without any adverse effects to her neurologic status. Magnetic resonance imaging of the brain was performed before and after the course of therapy and revealed no change in the white matter lesions visualized on spin echo images. PMID- 11940907 TI - ECT in the Treatment of Organic Psychosis in Huntington's Disease. AB - Huntington's disease (HD) is a progressive neurological disorder characterized by choreiform movements, dementia, and psychiatric symptoms. The psychiatric symptoms often lead to long-term psychiatric hospitalization. We report the case of a 49-year-old woman with HD of 1 year's duration who presented with an organic psychotic syndrome with voices telling her to kill herself or her children. The psychosis was refractory to neuroleptic treatment (up to 1,250 mg chlorpromazine equivalents per day). Depressive symptoms emerged but were neither of a magnitude nor of a duration to suggest a major affective disorder. Nonetheless, we treated the patient empirically with antidepressants, which had to be discontinued because of symptomatic orthostatic hypotension. Based on the persistent psychosis with suicidal and homicidal ideation and possible long-term state psychiatric hospitalization, we recommended electroconvulsive therapy (ECT). ECT was successful in treating the psychosis as well as the depressive symptoms. There was no worsening of the patient's dementia and only a slight progression in her choreiform movements. The authors question whether ECT may be of benefit in treating the psychosis of HD in the presence or absence of depressive symptoms and call for further research to determine if the use of ECT in HD patients with psychosis could prevent, or at least delay, placement in a long-term psychiatric hospital. PMID- 11940909 TI - A Reply to Brumback. PMID- 11940908 TI - EEG Monitoring. PMID- 11940910 TI - Use of a Peripheral Nerve Stimulator with ECT. PMID- 11940911 TI - Training in Convulsive Therapy in Denmark. PMID- 11940912 TI - ECT in Schizophrenia. PMID- 11940913 TI - Is ECT Usage Decreasing? PMID- 11940914 TI - Memory Disturbance After ECT in Low-Pressure Narcosis: A Study of Anterograde and Retrograde Amnesia. AB - In order to examine the significance of blood pressure elevation during electroconvulsive therapy (ECT) for posttreatment cognitive impairment, routine treatments were compared with treatments where the blood pressure elevation was attenuated by administration of trimethaphan (Arfonad). As a reliable, valid, and sensitive measure of cognitive impairment, the forgetting score in four memory tests was used. Both retrograde and anterograde amnesic effects were examined. In spite of a substantial attenuation of the blood pressure response, there was no decrease of the amnesic effects. It is concluded that low-pressure narcosis is not successful in alleviating memory disturbance after ECT. There is no support to the concept that the blood pressure elevation is relevant to the memory disturbance. Since there was a tendency toward decreased seizure duration, low pressure narcosis may be inappropriate because it counteracts a physiological compensatory mechanism to meet the increased metabolic demands during the seizure. PMID- 11940915 TI - Electroconvulsive Therapy and Neuroleptic Medication in Therapy-Resistant Positive-Symptom Psychosis. AB - Eight patients with persistent psychosis and positive symptoms, who had not improved with at least two courses of neuroleptic medications, were treated with electroconvulsive therapy (ECT) combined with an antipsychotic medication. Seven of the eight patients improved; the improvement was sustained in five patients after 6 and 12 months. ECT and neuroleptic drugs appear to be synergistic in antipsychotic activity. This enhancement may result from greater brain levels of neuroleptic drugs secondary to increased permeability of the blood-brain barrier accompanying seizures. Our experience is consistent with other recent clinical reports, and encourages trials of ECT combined with neuroleptic medications in therapy-resistant psychotic patients, including those classified as having schizophrenic disorders. PMID- 11940916 TI - Monoamine Oxidase Inhibitors in General Anesthesia: A Reevaluation. AB - A controlled study on the cardiovascular safety of monoamine oxidase inhibitors (MAOI) during general anesthesia for electroconvulsive therapy (ECT) was completed. No serious changes in blood pressure, heart rate, or ectopic beats were noted in either the MAOI or control groups during ECT. Based on pharmacological data, a review of the literature, and these preliminary results, the recommended 2-week discontinuation of MAOI before ECT appears unwarranted. Clinical guidelines for the concomitant use of MAOI and general anesthesia are offered. PMID- 11940917 TI - Electroconvulsive Therapy: A German Perspective. AB - A survey was undertaken in 1986 to provide reliable information on the present status of electroconvulsive therapy (ECT) in the Federal Republic of Germany (FRG). ECT was used in 26% of the state mental hospitals, 40% of the psychiatric departments of general hospitals, and in 78% of university hospitals. The number of treatment courses applied was low, with about 500 patients receiving ECT in 1985. The indications for ECT and patterns of administration appeared similar to those of other countries, although unilateral electrode placement was prescribed infrequently. The low use of ECT in the FRG (0.08/10,000 population) cannot be related to the attitudes of senior psychiatrists toward ECT, because the attitudes appear to be rather favorable. The majority maintained that indications for ECT exist and that every psychiatric hospital with comprehensive treatment facilities should offer this treatment. ECT usage seems to be partly determined by nonmedical factors, in particular, the recent attacks against psychiatry and ECT in the media. To correct the consequences of such attacks, the merits of ECT should become better known. PMID- 11940918 TI - Short-term Course and Outcome of Patients Treated with Electroconvulsive Therapy: Irish and British Surveys Compared. AB - Comparison of two independently conducted electroconvulsive therapy (ECT) surveys in Great Britain and Ireland shows remarkable uniformity of opinion and practice in the selection of patients for treatment, treatment technique, and clinical outcome 14 and 28 days after completion of a course of treatment. Short-term outcome reported by a self-selected group of treating psychiatrists in each survey was remarkably similar. Between 75% and 80% of patients were satisfied with the results of ECT as were their treating psychiatrists. The data provide information hitherto unavailable from controlled efficacy studies and indicate the short-term course and outcome of patients treated with ECT in contemporary clinical practice. PMID- 11940919 TI - ECT for Post-stroke Depression: Beta Blockade to Modify Rise in Blood Pressure. AB - A 60-year-old normotensive woman, who 3 years previously had had a cerebral haemorrhage, developed a severe depressive illness that failed to respond to tricyclic antidepressants and lithium, alone and combined. Electroconvulsive therapy (ECT) was administered using intravenous beta-blockade with practolol and atenolol prior to induction of anaesthesia to prevent the surge in blood pressure normally seen following the convulsion. The blood pressure did not rise and her depression responded to a series of nine ECT treatments. There were no associated complications. PMID- 11940920 TI - ECT and Anticoagulation. AB - Electroconvulsive therapy (ECT) is an effective and safe treatment for depression. Anticoagulation therapy is used to reduce morbidity or mortality from stroke or recurrent pulmonary embolus in various cardiac and vascular conditions. We report in detail the successful ECT treatment of a patient suffering from major depression with psychotic features who continued warfarin (Coumadin) anticoagulant therapy. PMID- 11940921 TI - A Response to Overall and Rhoades Regarding Their Comment on the Efficacy of Unilateral vs. Bilateral ECT. PMID- 11940922 TI - Clarification of Issues Related to Meta-Analysis for Studies of Convulsive Therapy. PMID- 11940923 TI - ECT and DST. PMID- 11940924 TI - Anticoagulants and ECT. PMID- 11940925 TI - Maintenance ECT and Affective Disorders. PMID- 11940926 TI - Continuation Therapy After ECT for Delusional Depression: A Naturalistic Study of Prophylactic Treatments and Relapse. AB - The study examines the naturalistic clinical course of 21 patients with delusional depression following their acute response to ECT. Their course is contrasted with that of 31 patients with delusional depression who acutely responded to medication treatment. They were treated for a minimum of 6 months (mean 3.5 years) with various continuation medications. Delusional depression is associated with a high rate of relapse, and some form of continuation therapy is clearly required. Akin to the acute treatment studies of delusional depression, prophylaxis with antidepressants alone is inadequate in preventing relapse in delusional depression patients. PMID- 11940927 TI - Maintenance ECT: A Survey of Practice (1986). AB - A questionnaire on the use of maintenance or continuation ECT was mailed to 173 practitioners of ECT. Of 86 respondents, 51 reported they used maintenance therapy, and 35 did not. Users came from 24 states, with five states accounting for more than half. Of practitioners giving maintenance ECT, each treated a median of three patients annually between 1981 through 1984, with a range from zero to "thousands." Frequency of treatments is determined empirically, with most treatments given monthly. Memory impairment is the most commonly cited risk. Depression is the diagnosis in 90% of the patients, with schizoaffective, schizophrenic, and manic patients equally comprising the remainder of the patients. The longest series reported for any one patient is 2,400 treatments. PMID- 11940928 TI - Isoflurane Anesthesia Therapy: A Replacement for ECT in Depressive Disorders? AB - Isoflurane anesthesia therapy (IAT) has been suggested to have persistent antidepressant activity. In a replication trial, IAT was performed in six patients who had a recurrence of a severe depressive episode, previously treated with ECT. In 21 of 26 IAT sessions, an isoelectric electroencephalogram lasting between 5 and 12 min was recorded. Clinical antidepressant activity was not observed. It is concluded that IAT is not an effective replacement for ECT in the treatment of major depressive disorders. PMID- 11940929 TI - Electroconvulsive Therapy in the Neuroleptic Malignant Syndrome. AB - A case of neuroleptic malignant syndrome (NMS) successfully treated with electroconvulsive therapy (ECT) is presented and the literature on this subject is summarized. The possibility of an etiologic link between catatonia related to a psychiatric disorder and NMS is explored. Possible relationships between NMS and lethal catatonia are explored. ECT may be a useful treatment in selected cases of NMS, especially those with evidence of a preexisting ECT responsive disorder. PMID- 11940930 TI - Modified ECT Using Succinylcholine After Remission of Neuroleptic Malignant Syndrome. AB - The authors report two patients with a history of neuroleptic malignant syndrome (NMS) who were treated with ECT using succinylcholine as muscle relaxant, without ensuing complications. Most, though not all, of the evidence suggests that NMS is primarily central in origin in contrast to malignant hyperthermia, which is due to a genetic defect in skeletal muscle. Regardless of the theoretical debate over the origin of the high temperatures noted in NMS, reports to date suggest that modified ECT using succinylcholine can be safely used after remission of an episode of NMS. PMID- 11940931 TI - Electroconvulsive Therapy in Acute Life-Threatening Catatonia with Associated Cardiac and Respiratory Decompensation. AB - A 61-year-old white female patient with known endogenous depression developed acute life-threatening catatonia. Associated cardiac and respiratory insufficiency prevented drug therapy. The patient was successfully treated with ECT. PMID- 11940932 TI - LSD-Like Flashbacks Associated with ECT. AB - We report the occurrence of spontaneous LSD (lysergic acid diethylamide)-like flashbacks after seven nondominant unilateral electroconvulsive therapy (ECT) treatments in a depressed patient with a history of hallucinogen abuse and flashbacks in the distant past. The similarity of the patient's reported experiences following ECT to the characteristic descriptions of LSD flashbacks is discussed. The differential diagnosis of LSD flashbacks in this case, and speculations regarding their origin, are also considered. PMID- 11940933 TI - Electroconvulsive Therapy in a Depressed Patient with a Functioning Ventriculoatrial Shunt. AB - We report a depressed man, with a ventriculoatrial shunt for normal pressure hydrocephalus, who was successfully treated with electroconvulsive therapy (ECT). The management of patients who have brain shunts and who are referred for ECT is discussed. PMID- 11940934 TI - Wave-Form and Clinical Efficacy. PMID- 11940935 TI - Lithium and ECT Interaction. PMID- 11940937 TI - Malpractice Insurance and Consent Regulations: Wisconsin Experience. PMID- 11940936 TI - A Reply to El-Mallakh. PMID- 11940938 TI - Fifty Years of Electroconvulsive Therapy. PMID- 11940939 TI - The Origins of Electroconvulsive Therapy (ECT). AB - Prior to the 1930s, the prime mode of treatment for psychiatric outpatients was psychoanalysis. Little could be done for inpatients, other than provide sedation and social support. In the 1930s, four major somatotherapies, all interventionist in technique, were developed: insulin coma therapy, Metrazol convulsive therapy, lobotomy (psychosurgery), and electroconvulsive therapy (ECT), the only one of these therapies still in use today. This paper focuses on the development of ECT by Ugo Cerletti and Lucio Bini at the Clinic for Nervous and Mental Disorders in Rome in 1938. The first electroshock treatment with humans is discussed in detail and the export of ECT to North America is described. Fifty years after the first treatment, ECT remains a controversial method of psychiatric treatment. PMID- 11940940 TI - Interview with Lothar Kalinowsky, M.D., October 8, 1987. PMID- 11940941 TI - An Eyewitness Account of the Discovery of Electroshock. AB - The author was one of the pupils of U. Cerletti and his assistant for almost 20 years. He worked with his master in the discovery of electroshock and in its first application in man in 1938. The author has thought it useful to publish his own testimony for the purpose of making the truth about the events known-the same truth that has often been questioned. PMID- 11940942 TI - The First ECT Devices. AB - The earliest electroconvulsive therapy (ECT) equipment was surprisingly advanced in its characteristics. Designed by some of the leading clinical scientists of the day in conjunction with creative engineering entrepreneurs, many devices contained features as advanced as are now in contemporary devices. A review of the development of ECT devices is of historical interest and it offers a fresh glimpse into technical issues that occupy recent interest. PMID- 11940943 TI - Effect of ECT on Cerebral Blood Flow in Melancholia Assessed with SPECT. AB - Ten endogenously depressed patients were studied before, during, and after electroconvulsive therapy. The nature and severity of depression was measured using the Newcastle Rating and Hamilton Depression Scales. The mean number of treatments administered was 12, and all patients recovered. Regional cerebral blood flow was studied using single photon emission computerized tomography (SPECT) with inhalation of (133)Xe. The mean values dropped by 8% after session three and a further 13% after the last session. The flow values of untreated patients was significantly higher than those found in 10 healthy volunteers. The values in patients following the last treatment session did not differ from those in the control group. There were no changes in the regional distribution of blood flow between controls and among patients in the three situations studied. No correlation was found between the physiological data and single items from the rating scales used. Increased cerebral blood flow values found in the treatment situation are due to the presence of depressive illness as such and not to agitation or anxiety. The parallel between flow data and clinical data points to the normalization taking place after, and not during, electroconvulsive therapy. PMID- 11940944 TI - Unilateral ECT in the Treatment of Manic Episodes. AB - It has been suggested that unilateral electroconvulsive therapy (ECT) is less effective than bilateral ECT in manic patients. In a pilot study, six medication resistant manic patients received either left or right unilateral ECT. In an ongoing study, 14 similar patients were randomly assigned to receive left unilateral, right unilateral, or bilateral ECT. Across both studies, a therapeutic equivalence for unilateral and bilateral ECT was found. Possible reasons for disparate findings with respect to the efficacy of unilateral ECT in mania are discussed. PMID- 11940945 TI - Mania as a Side Effect of Electroconvulsive Therapy. AB - Thirty-two endogenous depressed patients (RDC) were treated with electroconvulsive therapy. Four patients (12.5%) developed a transient manic reaction; in two cases, this reaction occurred in mid-depression. Mania is not a generally recognized side effect of ECT; we detail the clinical characteristics, but fail to define clinical predictors of vulnerability. PMID- 11940946 TI - Monitoring Electroconvulsive Therapy by Galvanic Skin Response. AB - Galvanic skin response (GSR) was assessed during 10 electroconvulsive therapy (ECT) sessions with five patients to evaluate its clinical utility in monitoring ECT-induced seizures. While GSR change was consistently observed in all instances, its duration persisted far longer than did seizure activity as measured by observed tonic-clonic movements. Accordingly, and in contrast to a prior report, GSR cannot be recommended as an accurate and reliable method of monitoring ECT. PMID- 11940947 TI - Reporting of Technical Parameters in ECT Publications: Recommendations for Authors. PMID- 11940949 TI - Might "Isoflurane Narcotherapy" Replace ECT? PMID- 11940948 TI - Electroconvulsive Therapy in a Patient with Mania, Parkinsonism, and Tardive Dyskinesia. AB - The authors describe a patient in whom ECT was associated with recovery from a manic episode, improvement in coexistent parkinsonism, and a marked change in the anatomic distribution of dyskinetic movements. PMID- 11940950 TI - Schizophrenia and ECT. PMID- 11940951 TI - Convulsive Therapy in the Treatment of Mania: McLean Hospital 1973-1986. AB - The authors completed a retrospective chart review of the records of all patients identified with diagnoses of mania and schizoaffective disorder, manic type, who underwent electroconvulsive therapy between the years 1973 and 1986 at McLean Hospital. Ten of 18 manic patients (56%) and 3 of 9 schizoaffective patients (33%) experienced meaningful clinical benefit. The authors report the correlation of treatment and patient factors with outcome and review the literature on the convulsive therapy of mania. PMID- 11940952 TI - Convulsive Therapy and the Course of Bipolar Illness, 1940-1949. AB - A total of 93 bipolar patients admitted in the 1940s were divided into those who received convulsive therapy and those who did not. Clinically, the two groups were similar. Those who received convulsive therapy were discharged back home more frequently, indicating the efficacy of the treatment. Those who received convulsive therapy showed more episodes, both manias and depressions, in the course of their illnesses. The reason for this difference in course of illness is not known, but deserves further exploration. The explanation may involve either biological factors or social circumstances related to a successful treatment outcome. PMID- 11940953 TI - Disparate Biochemical Actions of Electroconvulsive Therapy and Antidepressant Drugs. AB - Electroconvulsive therapy (ECT) effects on monoamine transmitter metabolites in the cerebrospinal fluid (CSF) were evaluated in three patients after completion of a course of bilateral or unilateral ECT. Each patient had earlier undergone an unsuccessful trial with an antidepressant drug. Despite the disparate nature of the basic pharmacology of the antidepressant drugs used, common chronic effects were observed in the CSF, with reductions in 3-methoxy-4-hydroxyphenylglycol (MHPG) and 5-hydroxyindoleacetic acid (5-HIAA) concentrations in all patients despite lack of therapeutic response. Homovanillic acid (HVA) changes were inconsistent. After ECT, however, no CSF changes were observed in the one nonresponder to that treatment. The two ECT responders showed marked increases in CSF 5-HIAA and HVA over their respective baselines, with an elevation in MHPG in one patient only. Further study of the mechanisms of action of ECT should focus on the serotonin and dopamine systems and on the differences between responders and nonresponders. PMID- 11940954 TI - Serotonin and Electroconvulsive Shock-Induced Prolactin Release. AB - Baseline and post-electroconvulsive shock (ECS) prolactin levels were determined in serum samples from 13 rabbits, with and without pretreatment with the serotonin-2 (5-HT(2)) receptor blocker ketanserin. Without ketanserin, ECS did not raise serum prolactin levels over baseline. After ketanserin, baseline serum prolactin levels were 50% lower and post-ECS prolactin levels were twice this new baseline level and close to post-ECS prolactin levels without ketanserin. These observations suggest that serotonergic neurotransmission potentiates stress induced prolactin secretion, but contributes little to ECS-induced prolactin release. PMID- 11940955 TI - Patients' Views on Electroconvulsive Therapy on Completion of a Course of Treatment. AB - Patients' attitudes towards electroconvulsive therapy (ECT) on their completion of a course of treatment were studied using a semi-structured questionnaire. Twenty-six patients were seen before the junior doctors were trained in ECT administration and 28 patients afterwards. Most patients' attitudes were favourable, although very few fully understood the treatment. Training the junior doctors had no effect on patients' attitudes, but was associated with a shorter mean course length. ECT was acceptable to most patients who received it. PMID- 11940956 TI - Relative Efficacy of Unilateral and Bilateral Electroconvulsive Therapy in Melancholia. AB - We compared the efficacy of unilateral nondominant (n = 15) and bilateral (n = 31) electroconvulsive therapy (ECT) in melancholia in 46 consecutive, nonrandomly assigned medication-free patients with endogenous depression. Seizure duration was recorded and, if a seizure lasted <25 s, the stimulus was immediately readministered. There were more missed or brief seizures that required restimulation with unilateral ECT than with bilateral ECT. After five treatments, blind assessments on the Hamilton Rating Scale for Depression showed a 57% improvement in the bilateral group as compared with a 19% improvement in the unilateral group. The unilateral group received more total treatments (mean = 9.9) than did the bilateral group (mean = 7.7). At the conclusion of all ECT treatments, 72% of patients receiving bilateral ECT were substantially improved and 14% partially improved, compared with 32% of patients receiving unilateral ECT with substantial improvement and 30% partial improvement. This difference between the two groups, however, had disappeared by the time of discharge. Although unilateral ECT may induce fewer side effects, we find bilateral treatments to be more efficacious, at least on a short-term basis. PMID- 11940957 TI - Factors Influencing Number of Treatments and Seizure Duration in ECT: Drug Treatment, Social Class. AB - This investigation studies number of treatments and seizure duration during a course of unilateral electroconvulsive therapy (ECT) in 82 inpatients with major depressive disorders. Therapy with antidepressants or benzodiazepines preceding ECT was neither related to number of treatments nor to seizure duration. An average dose of methohexital higher than 1.2 mg/kg body weight was associated both with an increased number of treatments and a shorter average seizure duration. Concurrent therapy with neuroleptics was related to a reduced number of treatments. Patients in social class I, more often than others, received fewer treatments. PMID- 11940958 TI - Predicting the Outcome of Endogenous Depression Following Electroconvulsive Therapy. AB - Endogenous depression is known to be associated with good outcome following electroconvulsive therapy (ECT). In a double-blind, prospective study, we applied three clinical predictive indices and one diagnostic index to a cohort of 29 endogenous depressed patients, to obtain better predictors of outcome following ECT. The Newcastle Prognostic Index identified ECT responders with high specificity but low sensitivity; other indices, such as those described by Hobson (1953) and by Mendels (1967), were neither sensitive nor specific in predictive standards. If ECT-treated depressed patients are pre-selected for endogenous symptomatology, fresh clinical predictive indices need to be developed. PMID- 11940959 TI - Neuroleptic Malignant Syndrome, Malignant Hyperthermia, and ECT. PMID- 11940961 TI - Lithium-ECT Interaction. PMID- 11940960 TI - A Reply to Pearlman. PMID- 11940962 TI - A Reply to Pearlman. PMID- 11940963 TI - ECT for Parkinson's Disease? PMID- 11940964 TI - Medication Outcome in ECT-Resistant Depression. AB - The outcome of antidepressant treatment in 12 cases of electroconvulsive therapy (ECT)-resistant depression is presented. Eight patients had been refractory to a clinically adequate course of ECT (Hamilton Depression Scale improvement <20%) and four were partial responders (improvement 20-49%). All remitted completely on antidepressant medication within 2.2 +/- 1.1 (mean +/- SD) months of the ECT course. Remission was associated with clomipramine treatment (139 +/- 49.7 mg/day) in seven cases and maprotiline (125 mg/day) in one case. Four patients who did not respond to a tricyclic antidepressant alone remitted following supplementation (of clomipramine in 2 cases, clomipramine + haloperidol in 1 case, and imipramine in 1 case) with lithium carbonate. Although a delayed therapeutic response to ECT cannot be excluded, the results suggest that ECT may alter the sensitivity of refractory patients to antidepressant medication. PMID- 11940965 TI - Effects of Electroconvulsive Therapy on Brain Glucose Metabolism: A Preliminary Study. AB - We measured (15)O water uptake and [(18)F]fluorodeoxyglucose (FDG) uptake in four depressed individuals before electroconvulsive therapy (ECT) and 24 h after completion of a series of six to 11 bilateral ECT treatments. Studies of radioactive uptake were done using positron emission tomography, with FDG as a measure for glucose metabolism and with oxygen-15-labeled water as a measure for cerebral blood flow. The four patients showed decreased uptake of FDG in the frontal cortex after ECT treatment. Two of the three patients in whom (15)O water was measured showed decreased uptake in the frontal cortex after ECT. PMID- 11940966 TI - Monitoring Seizure Duration During Electroconvulsive Therapy. AB - Three alternative monitoring methods for assessing the duration of seizures during electroconvulsive therapy were studied. The mean integrated amplitude of the electroencephalogram, facial muscle electromyogram, and "cuff method" were compared with the reference single-channel unprocessed electroencephalogram in 78 sessions with 17 patients. The measures of seizure duration differed significantly (p < 0.001). The mean integrated electroence phalographic amplitude differences were small and an artifact of the sampling procedure. Larger, but clinically unimportant, discrepancies were obtained with the facial electromyogram amplitude. In contrast, there were marked differences between the electroencephalogram and the "cuff method," which suggest that the latter technique may be of limited usefulness. PMID- 11940967 TI - Reevaluation of ECT in Schizophrenia: Right Temporoparietal versus Bitemporal Electrode Placement. AB - Two groups of schizophrenic patients received either bitemporal or right temporoparietal electroconvulsive therapy (ECT), both without anesthesia. Each patient received six treatments, the first three at 2-day intervals and the next three at 4-day intervals. Psychiatric evaluations and cognitive tests done at baseline, and 7 and 20 days after therapy, found similar degrees of improvement and similar effects on memory in both groups, with improvement in 40% and impairment in 30% of the patients. Subjective forgetfulness is more often experienced by the patients who receive bilateral ECT. PMID- 11940968 TI - ECT in Post-Stroke Major Depression. AB - A depressed male patient, who had his first affective disturbance as a complication of a right parietotemporal cerebrovascular accident, was treated with unilateral, nondominant electroconvulsive therapy, administered ipsilateral to the lesion. This procedure produced adequate seizure duration with excellent clinical results and without new neurological or neuropsychological deficits. PMID- 11940969 TI - Seizure Elicitation Blocked by Pretreatment with Lidocaine. AB - A patient is reported in whom pretreatment with intravenous lidocaine resulted in failure to elicit a seizure even at stimulus intensity markedly greater than that used successfully at prior and subsequent treatments. Together with one other report, this experience suggests that lidocaine may prevent a seizure by raising the seizure threshold. An alternative explanation is proposed for the classic findings of Ottosson that lidocaine-modified seizures are reduced in therapeutic efficacy. PMID- 11940970 TI - Treatment of Phencyclidine-Associated Psychosis with ECT. AB - Phencyclidine (PCP) abuse usually causes a short, self-limited period of intoxication but may lead to a prolonged psychosis, poorly responsive to antipsychotic medications. Prior reports indicate that electroconvulsive therapy (ECT) is a safe, rapidly effective treatment in such cases, but longer-term outcome has not been studied. We report three cases of PCP-associated psychosis that did not respond to at least 2 weeks of antipsychotic treatment but subsequently responded rapidly to ECT. Follow-up at 14 to 21 months after treatment show that recovery was maintained except in one subject who relapsed after continuing to abuse PCP. ECT should be considered as a treatment early in the course of drug-associated psychoses in which an adequate trial of antipsychotic medication has not been effective. PMID- 11940971 TI - ECT in a Patient with Conversion Disorder. AB - A 61-year-old man with a severe conversion disorder with paralysis of his hand and consequent disuse atrophy was successfully treated with electroconvulsive therapy (ECT) when he was threatened with the loss of his hand. The experience of ECT in patients with conversion disorder is reviewed. PMID- 11940972 TI - Petit Mal-Grand Mal ECT: A Method to Induce Seizures Without Barbiturate Anesthesia. AB - Petit mal-grand mal (PM-GM) electroconvulsive therapy (ECT) is a technique developed by Impastato to elicit unconsciousness with a subconvulsive electrical stimulus, rather than with barbiturate anesthesia. Muscle relaxation is produced with succinylcholine chloride before stimulus is applied. The cases reported here illustrate applications of the technique to depressed patients with severe cardiac and pulmonary disease, and the use of PM-GM ECT in a patient in whom seizures could not be elicited by the usual ECT technique is described. PMID- 11940973 TI - Mania as a Side-Effect of ECT. PMID- 11940975 TI - Continuation Therapy with Antidepressants After Electroconvulsive Therapy. PMID- 11940974 TI - "Cuff" Monitoring, Osteoporosis, and Fracture. PMID- 11940976 TI - Effects of ECT on Polysomnographic Sleep: A Prospective Investigation. AB - The authors describe a pilot investigation using ambulatory polysomnography (PSG) to assess rapid eye movement (REM) latency in 11 depressed inpatients before and after a course of electroconvulsive therapy (ECT). Prior to beginning ECT, all subjects had REM latencies of 56 minutes or less (mean 22 min). A course of ECT was associated with clinical improvement in every patient and a statistically significant (p < 0.0002) increase of 67% in mean REM latency (37 min). However, individual patients showed marked variability in REM latency both during and after the course of ECT, and 7 of the 11 responders continued to exhibit shortened REM latencies of 34 min or less. Technical considerations related to PSG studies during ECT are reviewed and the clinical and theoretical implications of our findings are discussed. PMID- 11940977 TI - Clinical, Neuropsychological, and EEG Evidence for Mechanisms of Action of ECT. AB - Clinical, neuropsychological, and electroencephalogram (EEG) data from 150 ECT treated patients with affective disorders were analyzed to bear upon current hypotheses about neurophysiological mechanisms of action of electroconvulsive therapy (ECT). Comparisons of responders and nonresponders revealed little or no evidence to support kindling, sensitization, diencephalic stimulation, or anticonvulsant theories. Neuropsychological test data supported right hemispheric dysfunctions in affective illness which resolved more in ECT responders than in nonresponders. There were more impairments in dominant and bilateral hemispheric functions after successful than after unsuccessful ECT. The amount of EEG slowing and paroxysmal activity was not related to therapeutic response. Transient left sided EEG slowing was associated with positive ECT response whereas right-sided EEG changes were not. The data support the hypothesis that in patients with affective disorders, ECT restores hemispheric equilibrium with enhancement of right brain functions and dampening of the left. PMID- 11940978 TI - Electroconvulsive Shock Does Not Increase Plasma Insulin in Rats. AB - This study examines the time course of plasma insulin and glucose concentrations following a single electroconvulsive shock (ECS) and the last of 10 once-daily ECS in rats. In contrast to a report in humans, neither single nor repeated ECS leads to an increase of plasma insulin. There was only a slight increase in blood glucose concentrations following either treatment. PMID- 11940979 TI - A Double-Blind Comparison of Sinusoidal Wave and Brief-Pulse Electroconvulsive Therapy in Endogenous Depression. AB - In a double-blind prospective study, 29 endogenously depressed patients (RDC) were randomized into sinusoidal wave (SW) and brief-pulse (BP) electroconvulsive therapy (ECT) groups. Bilateral modified treatments were administered on alternate days, three per week, and the treatment variables of current dosage and seizure duration were monitored for each treatment. Significantly more patients responded to SW than to BP ECT, but a comparable number of treatments was required to produce this response in the two groups. There was no difference in clinical or treatment variables between the SW and the BP groups, nor between ECT responders and nonresponders, with the exception that the SW-treated patients received larger doses of current per treatment than did the BP patients. For endogenous depression treated with ECT, we conclude that cumulative seizure duration may not be a parameter of significance, that overall rate of recovery in ECT responders is independent of stimulus waveform, and that some depressives may respond to SW but not to BP ECT. We suggest that the antidepressant effect of the ECT seizure may be characterized by a therapeutic window in current requirements; alternatively, a putative response threshold (again in terms of current requirement) may exist, which is higher in some patients than in others. PMID- 11940980 TI - Electroconvulsive Therapy in Aarhus, Denmark, in 1984: Its Application in Nondepressive Disorders. AB - The indications for electroconvulsive therapy (ECT) are focused on one or a few treated subpopulations. We retrospectively reviewed all ECT treatment series in 1984 in Aarhus Psychiatric Hospital, Risskov, Denmark. Patients with diagnoses other than endogenous depression were successfully treated with ECT. The diagnoses included mania, manic-depressive mixed states, acute exacerbations in schizophrenia, reactive psychosis and delirium, most of whom were in a very severe condition. This experience is described. PMID- 11940981 TI - Electroconvulsive Therapy in Obsessive-Compulsive Disorder. AB - An open trial of electroconvulsive therapy was conducted in nine subjects who met DSM-III criteria for obsessive-compulsive disorder. There was an initial reduction in symptomatology that lasted from 1 to 4 months. Subjects who had less obsessive-compulsive (anankastic) personality traits responded better. There was also the post hoc observation that subjects who were agitated did better. We observed correlations between depression and interference scores, but not with symptom scores, suggesting that ECT has anti-obsessional activity. PMID- 11940982 TI - A Mechanism for the Efficacy of ECT in Parkinson's Disease. PMID- 11940983 TI - Tardive Dyskinesia and Electroconvulsive Therapy. AB - An elderly female patient with major depression and persistent tardive dyskinesia was treated with electroconvulsive therapy (ECT). In addition to complete recovery from depression, a significant improvement was also observed in her symptoms of tardive dyskinesia. Further study of the effects of ECT on tardive dyskinesia is warranted. PMID- 11940984 TI - Hemodialysis, Chronic Renal Failure, and ECT. PMID- 11940985 TI - Beta Blockade During ECT: A Cautionary Note. PMID- 11940986 TI - ECT Hypothesis: A Correction. PMID- 11940987 TI - ECT in the High-Risk Patient. PMID- 11940988 TI - Effect of a Single Nighttime Dose of Oxazepam on Seizure Duration in Electroconvulsive Therapy. AB - In a double-blind crossover study, 12 depressed inpatients receiving bilateral electroconvulsive therapy (ECT) were given 22.5 mg oxazepam or placebo (identical tablets, randomized order) the night before treatment. Seizure duration was measured using the cuff method and a total of 20 pairs of measurements were made. Mean duration after oxazepam administration was 0.9 s longer than after placebo; this difference was not statistically significant. The relevant lower one-sided 95% confidence limit was -4.1 s. Single administration of oxazepam, 22.5 mg, has little average effect on seizure duration in ECT. PMID- 11940989 TI - Electroconvulsive Therapy in the Medically Ill Elderly. AB - Treatment with electroconvulsive therapy (ECT) of medically ill geriatric patients with affective disorders is described. Of 135 patients to whom ECT was administered in 6 years, 55% were over 60 years of age. One-third (45) exhibited cardiovascular (62%), central nervous system (15%), or other medical conditions (22%) increasing the risks of the treatment. Treatment strategies are described. PMID- 11940990 TI - Age-Related Changes in Cardiovascular Function Associated with Electroconvulsive Therapy. AB - The relationship between hemodynamic changes induced by electroconvulsive therapy (ECT) and age was studied in 13 patients. Their average age was 43 years (range 20-64 years). Heart rate, arterial blood pressure, and rate-pressure product (RPP) all increased significantly following ECT. There was an inverse relationship between age and the magnitude of the chronotropic response, the percentage increase in diastolic and mean arterial blood pressure, and the RPP. Duration of the electroencephalographic and motor seizures was also inversely related to age. PMID- 11940991 TI - Brain Magnetic Resonance Imaging in Elderly Depressed Patients Receiving Electroconvulsive Therapy. AB - The authors review their experience with brain magnetic resonance (MR) imaging in elderly patients with depression referred for electroconvulsive therapy (ECT). A variety of brain changes were identified on the pre-ECT MR scans of these patients, including cortical atrophy, subcortical encephalomalacia, lateral ventricular enlargement, and lesions of the pons. The authors examine the effects of these preexisting brain changes on the therapeutic outcome and side effects from ECT. Pilot data suggest that these brain findings do not change during a course of ECT. The potential relationship of these brain changes to the development of affective disorder in the elderly is discussed and areas for future research are suggested. PMID- 11940992 TI - Cardiovascular Effects of ECT. PMID- 11940993 TI - Methohexital Dosage in Electroconvulsive Therapy. AB - A comparison of methohexital at 0.5 mg/kg versus 1 mg/kg iv as anesthesia for unilateral brief pulse electroconvulsive therapy showed no difference in systolic or diastolic blood pressure before, immediately after, or 5 or 10 min after the seizure. Mean seizure duration was not significantly shorter using the higher methohexital dosage. These results show that methohexital can be safely used in a broad dosage range without undue effects on blood pressure or seizure duration. PMID- 11940994 TI - Atropine in Electroconvulsive Therapy. AB - Nineteen patients with major depression were alternately given intravenous atropine or saline immediately prior to anesthesia for electroconvulsive therapy (ECT). Atropine increased the heart rate, reduced the number of dropped beats, and reduced the number of premature atrial beats. These features may be advantageous in patients with cardiac hypodynamic states presenting for ECT, that is, with bradycardia, bradyarrhythmia, or hypotension. However, as atropine also increased the cardiac work, we recommend that it not be given to patients with hypertension, tachycardia, or who are at risk for cardiac ischemia. PMID- 11940995 TI - Comparison of Atropine with Glycopyrrolate for Use in ECT. AB - The occurrence of undesirable events during the postictal electroconvulsive therapy (ECT) period was retrospectively compared between 29 patients who had received glycopyrrolate premedication 0.002 mg/lb i.v. and 25 patients who had received atropine 1.0 mg i.m. A separate comparison was made for 14 patients who had received ECTs with each of these premedications. A total of 952 ECTs were reviewed. In both comparisons supraventricular tachycardia was more common with atropine (p <.0001 with each), after approximately 13% of ECTs. Otherwise, postictal events with the two agents were indistinguishable; specifically, bradycardia and nausea were not more frequent with glycopyrrolate. PMID- 11940996 TI - Safety and Efficacy of ECT in Depressed Patients with Dementia: A Review of Clinical Experience. AB - The authors review the results of electroconvulsive therpay (ECT) in 135 cases of depression occurring in conjunction with organic dementia, subcortical leukoencephalopathy without dementia, and depressive dementia (22 cases). Overall, 86% had a positive therapeutic response to ECT, whereas 21% experienced significant cognitive or memory side effects, virtually all of which were transient and reversible. Forty-nine percent of the patients with organic or depressive dementias experienced improvement in cognitive or memory function consequent to ECT. PMID- 11940997 TI - ECT in the High-Risk Geriatric Patient. AB - Electroconvulsive therapy (ECT) is an important therapeutic modality in geriatric patients because its safety, high efficacy and rapid onset of action are suited to the special needs of this population. It appears that ECT can be effective in the acute treatment of depression, mania, paranoid disorders and various psychiatric syndromes occurring in the context of brain disease. Certain cardiovascular and neurological disorders or use of drugs increase the risk of ECT in geriatric patients. Clinical strategies for evaluating and reducing the ECT risk in such cases are presented and directions for future clinical research are discussed. PMID- 11940998 TI - Anesthesia for Electroconvulsive Therapy in Patients with Major Cardiovascular Risk Factors. PMID- 11940999 TI - Electroconvulsive Therapy in Geriatric Patients Undergoing Anticoagulation Therapy. AB - Antidepressant treatment of the depressed geriatric patient can be complicated by concomitant medical illness. We report the successful administration of electroconvulsive therapy (ECT) in three depressed geriatric patients who received the oral anticoagulant warfarin (Coumadin) for cardiovascular disease. The physiologic changes associated with modified ECT and risk factors for intracerebral hemorrhage in patients receiving anticoagulants are discussed. PMID- 11941000 TI - Nifedipene in the Treatment of Blood Pressure Rise After ECT. AB - Extreme rise of blood pressure may increase the risk associated with electroconvulsive therapy (ECT) in patients with cardiovascular disease, cerebrovascular disease, and aortic and intracranial aneurysms. A case is presented in which sublingual nifedipene was used in the treatment of a high-risk patient with severe rise of blood pressure. The therapeutic alternatives of managing severe ECT-related blood pressure increases are presented. PMID- 11941002 TI - Psychiatric Malpractice and ECT: A Review of National Loss Experience. PMID- 11941001 TI - Reversible and Irreversible Dementia. PMID- 11941003 TI - Reliability of the Application of a Kinetic Model of Hormone Release: Prolactin and Oestrogen-Stimulated Neurophysin After Electroconvulsive Therapy. AB - Pituitary hormone release after electroconvulsive therapy (ECT) may provide important information about certain of the effects of ECT on neurotransmitter function. Previously we quantified pituitary hormone release by the increase in the area under the hormone concentration/time curve. Swartz has rightly pointed out that this method ignores the rate of elimination of hormone and cannot be a measure of total hormone released. Swartz has proposed a kinetic model of hormone release, and in this paper we assess the reliability of parameters derived from it when applied to our own data on the effects of ECT on the release of prolactin and oestrogen-stimulated neurophysin. The deviations between observed and predicted values were small when only five post-ECT samples were used. Contrary to expectations, the goodness of fit did not improve when the number of sampling points was increased to nine after ECT. It was possible to fit a unique curve to the observed data in only 28 of 54 data sets. The reasons for these findings are discussed. PMID- 11941004 TI - Clinical Features and Predictors of Postictal Excitement. AB - In a retrospective case control study, clinical features of postictal excitement following electroconvulsive therapy (ECT) were examined in 24 patients and 24 matched controls. The two groups did not differ in clinical outcome, degree of pretreatment agitation, anesthetic or succinylcholine dose, number of ECTs received, mean seizure duration, number of titrated treatments with subconvulsive stimulation, or type of EEG ending. Patients who became excited early in the treatment course rarely manifested this phenomenon at all subsequent treatments. Within the "excited" group, a minority of patients became excited following the first ECT, and only 34.3% of their treatments resulted in postictal excitement. It may not be necessary to routinely use postseizure benzodiazepines in patients who have once exhibited a postictal excitement. PMID- 11941005 TI - Prolactin Response to Pentylenetetrazol (Cardiazol) Convulsive Therapy. AB - Epileptic grand mal seizures as well as electroconvulsive therapy (ECT) induce a transient robust prolactin hypersecretion. Similar prolactin response has been demonstrated following pentylenetetrazol (Cardiazol)-induced seizures in two schizophrenic female patients. A slight increase in cortisol secretion but no change in thyroid stimulating hormone and growth hormone levels suggest that the prolactin response is a specific hormonal change during convulsive treatments. PMID- 11941006 TI - Effect of Single and Repeated Electroconvulsive Shock on Body Temperature in Mice. AB - The time course of rectal temperature in mice was measured following a single and the last of 10 once-daily electroconvulsive shocks (ECS). In addition, temperature measurements were carried out following a single ECS 1 and 2 weeks after a course of 10 once-daily ECS treatments. After a single ECS, the animals showed a decrease of 2.3 +/- 0.3 degrees C, lasting for at least 90 min. Following the last of 10 ECS, the maximum response was markedly attenuated to 0.7 +/- 0.1 degrees C, and this attenuated response was present at least 1 week later. This effect may be mediated by alterations in postsynaptic alpha(2) adrenoceptor or presynaptic 5-HT(1A) receptor sensitivity, or may reflect a more fundamental resetting of temperature mechanisms. PMID- 11941007 TI - Dopaminergic Effects of Repeated Electroconvulsive Shocks. AB - Dopamine (DA) autoreceptor and postsynaptic receptor changes following repeated electroconvulsive shocks (ECS) were investigated in rats using the indices of low and high-dose apomorphine-induced motility responses. Repeated ECS produced no changes in the DA autoreceptors; however, enhanced postsynaptic receptor-mediated responses were observed, suggesting increased sensitivity of the DA postsynaptic receptors. PMID- 11941008 TI - Maintenance ECT in Coexisting Affective and Neurologic Disorders. AB - To the numerous reports of the safety and efficacy of electroconvulsive therapy (ECT) in acute depressive episodes in patients with coexisting neurologic illness, the authors report their experience with two patients treated with maintenance ECT for more than 2 years. The authors find maintenance ECT useful in ameliorating the depressive illness without worsening the neurologic illness. PMID- 11941009 TI - Electroconvulsive Therapy and Asymptomatic Meningioma. AB - An elderly depressed female patient, who tolerated antidepressant drugs poorly, was found to have a small and asymptomatic meningioma before she was given electroconvulsive therapy (ECT). She received seven bilateral ECTs without neurological deterioration and had a good clinical result. PMID- 11941010 TI - Safety and ECT Stimulus Electrodes: I. Heat Liberation at the Electrode-Skin Interface. AB - Calculations reveal that the electroconvulsive therapy (ECT) stimulus cannot cause burn injury to brain tissue, but poor electrode-skin contact is a risk for skin burn. This article reviews factors that influence skin temperature, including electrical charge, skin impedance, topographic conformity between electrode and skin, electrode surface area, and thermal diffusivity of electrode material. PMID- 11941011 TI - Safety and ECT Stimulus Electrodes: II. Clinical Procedures. AB - The accidental risks and problems associated with the stimulus in electroconvulsive therapy (ECT) include skin burn, inadvertent exposure to the stimulus, short-circuit of the stimulus with ineffectual delivery, and accidental mislocation. These problems and aspects of their prevention are reviewed. PMID- 11941013 TI - A Reply to Sackeim et al. PMID- 11941012 TI - Should Tricyclic Antidepressants or Lithium Be Standard Continuation Treatment After ECT: An Alternative View. PMID- 11941014 TI - Comparison of Brief-Pulse and Sine Wave ECT Stimuli. PMID- 11941016 TI - Low-Dose Caffeine in ECT. PMID- 11941015 TI - Anomalous Patterns of ECT-Induced Hormone Release: Commentary on the Article by McGuire et al. PMID- 11941017 TI - When Is an ECT Responder, an ECT Responder? PMID- 11941018 TI - A Reply to Andrade and Gangadhar. PMID- 11941019 TI - Cardiologist as Alternative to Anesthesiologist for ECT? PMID- 11941020 TI - The Unique Contributions of ECT to Understanding the Pathophysiology and Treatment of Affective Disorders. PMID- 11941021 TI - Implications of Clinical Spectrum for Mechanisms of Action: ECT and Antidepressants Reconsidered. AB - The similarities and differences between electroconvulsive therapy (ECT) and antidepressant medications are reviewed with respect to their clinical indications. The implications of the overlap and divergence in their spectra of clinical efficacy are discussed in reference to mechanisms of action. The hypothesis is offered that ECT has multiple mechanisms of action, which differ depending on the clinical syndrome being treated. From this perspective, ECT may share similar mechanisms of action with other agents that are also effective in treating the specific syndrome. Thus, although ECT may result in a plethora of neurobiological effects, depending on the clinical syndrome, specific and differing subsets of neurobiological changes are relevant to therapeutic action. PMID- 11941022 TI - Mechanisms of the Antimanic Effect of Electroconvulsive Therapy. AB - The available evidence indicates that electroconvulsive therapy (ECT) has a marked and specific antimanic effect. Data on change in seizure threshold during ECT and acute effects of ECT on cerebral blood flow are presented in support of the view that the therapeutic effects of ECT are related to its anticonvulsant properties in manic patients. Specifically, an antimanic effect of ECT was associated with a significantly greater increase in seizure threshold during bilateral ECT and, across treatment conditions, with a significantly greater reduction in cerebral blood flow acutely following ECT. A low initial seizure threshold during bilateral ECT was found to be associated with a poor therapeutic response. This, along with earlier findings in depressed patients, suggests that heterogeneity of the disease processes underlying mood disorders may be critical for an understanding of the mechanisms of action of ECT. PMID- 11941023 TI - Lateralized Hemispheric Mechanisms and the Antidepressant Effects of Right and Left Unilateral ECT. AB - This article critically reviews the six controlled studies of the antidepressant effects of right unilateral versus left unilateral ECT (R-ECT, L-ECT), contrasting them with results recently obtained in double-blind, random assignment comparison of the two methods, using a square-wave stimulus, widely spaced temporoparietal electrode placement, and markedly supra-threshold stimulus charge. The author challenges the prevalent belief in a therapeutic advantage of R-ECT versus L-ECT, proposing instead the contrary case for a therapeutic advantage of L-ECT, based on the interaction of a hypothesized interhemispheric seizure threshold difference and a maximally efficient method of stimulus delivery. PMID- 11941024 TI - Neuropharmacological Aspects of ECT: In Search of the Primary Mechanism of Action. AB - Since its first use in humans 50 years ago, electroconvulsive therapy has been a controversial topic. Nevertheless, a great many experimental and clinical data have been collected. Much of it strengthens the notion that ECT is a more specific therapy than it appears at first sight. The pharmacological effects of ECT seem to be directed toward changes in signal transduction chain processes in the synapse and on the postsynaptic cell membrane. Some aspects of this have been well investigated, especially the effects of ECT on membrane receptors, whereas others have only recently evolved. This article discusses some general principles and conclusions and focuses on some very recent data acquired with new laboratory techniques, such as in vivo dialysis and molecular biology. An attempt is made to interpret these new data and to integrate them in previous results. PMID- 11941025 TI - Modulation of Endogenous Opioid Systems by Electroconvulsive Shock. AB - Since the discovery of opioid peptides, the brain opioid system has been implicated in the pathophysiology of a spectrum of mental disorders, including depression and epilepsy. For example, a growing body of evidence demonstrates that these neuropeptides are activated by seizures. Specifically, using electroconvulsive shock, it has been possible to describe an array of antinociceptive, autonomic, behavioral, biochemical, and electroencephalographic responses that appear to be mediated by endogenously activated opioids. A primary role for opioid peptides as neuromodulators of postictal seizure arrest and refractoriness is now recognized, and the existence in the central nervous system (CNS) of an endogenous anticonvulsant substance activated by electroconvulsive shock (ECS) has been determined. This review focuses on the more recent developments regarding ECS-induced modulation of brain opioid systems. The ability of ECS to alter opioid receptors, to influence the release and biosynthesis of the various opioid peptides, and to activate endogenous anticonvulsant mechanisms will be addressed. PMID- 11941026 TI - Perspectives on the Mechanism of Action of Electroconvulsive Therapy: Anticonvulsant, Peptidergic, c-fos Proto-oncogene Effects. AB - Although the mechanism of action of electroconvulsive therapy (ECT) in affective illness has remained elusive, it is hoped that the consideration of mechanisms underlying the anticonvulsant efficacy of ECT will provide new insights into its biochemical and neuroanatomical substrates. In the amygdala-kindling model, electroconvulsive seizures (ECS) inhibit both the development and completed phases of kindled seizure evolution, and therefore, ECS is a more potent anticonvulsant modality than carbamazepine, which inhibits only completed kindled seizures. Carbamazepine is increasingly recognized for its acute and prophylactic efficacy in bipolar affective illness. Thus, comparing and contrasting effects of ECS and carbamazepine may provide insights into overlapping mechanisms of anticonvulsant and psychotropic action. Anticonvulsant effects of ECS have been most closely linked to endogenous opiate substances, perhaps acting on delta opiate receptors, but a wide variety of other neurotransmitter and peptidergic effects are also potential candidates. Electroconvulsive seizures in mice activate the proto-oncogene c-fos in many discrete areas of brain, including a variety of limbic sites, the ventromedial nucleus of the hypothalamus, and the cerebellum. As such, c-fos induction may provide both an anatomical map of areas potentially activated by ECS and a potential mechanism for initiating a sequence of events that may be important to the mechanism of action of ECT. Although the anticonvulsant effects of ECT may ultimately prove to be separable from those mediating its therapeutic effects in affective illness, seizures and anticonvulsant effects provide easily measurable endpoints for preclinical and clinical studies. Given this clarity of effect, potential anticonvulsant mechanisms can rapidly be identified, enabling direct testing of whether or not these same mechanisms are also critical to the therapeutic effects of ECT in affective illness. PMID- 11941027 TI - A Neuroendocrine View of ECT. AB - A neuroendocrine hypothesis to explain the clinical activity of convulsive therapy is described. The hypothesis is based on the diabetes/insulin model and suggests that hypothalamic dysfunction with an insufficiency of a mood maintaining peptide is the basis for affective disorders. Repeated seizures enhance the production and release of the hypothalamic peptide antidepressin-the active principle that relieves both neuroendocrine and behavioral abnormalities. PMID- 11941028 TI - An Adequate Treatment? PMID- 11941029 TI - Efficacy of ECT in Psychotic and Nonpsychotic Depression. AB - The authors examined reports addressing the question of whether electroconvulsive therapy (ECT) is more effective in psychotic or nonpsychotic depressive disorders. Most ECT studies did not consider the issue directly, and those that did had varying methodologies and results. The authors then analyzed the data of 33 depressed patients to clarify the differential response to ECT in 13 psychotic and 20 nonpsychotic patients. The subjects were also examined as to type of electrode placement. ECT produced a greater reduction in Hamilton Depression Rating Scale scores in the psychotic depressives as compared to the nonpsychotics after an average of 9.2 treatments. The two patients who were nonresponders to ECT were both nonpsychotic. PMID- 11941030 TI - Efficacy and Safety of Combined ECT and Tricyclic Antidepressant Drugs in the Treatment of Depressed Geriatric Patients. AB - Clinical outcome of 84 elderly unipolar depressed patients who had received electroconvulsive therapy (ECT), with or without concurrent tricyclic antidepressant medication, was reviewed retrospectively to discover if predictions based on preclinical studies of increased efficacy with combined treatment would be supported. Global outcome at discharge was improved for those patients who had received tricyclic antidepressant drugs concurrent with ECT. They also required fewer ECT treatments to achieve that improvement. Total seizure times were markedly reduced for the combined therapy group, which was an unexpected finding. PMID- 11941031 TI - Continuation Therapy for Depression Using Outpatient Electroconvulsive Therapy. AB - This naturalistic study followed the course of 27 patients who received continuation therapy with outpatient electroconvulsive therapy (ECT) after responding well to an index course of ECT for major depression. The continuation outpatient ECT protocol consisted of weekly treatments for the 1st month after remission, followed by a gradual tapering to monthly treatments. All patients were free of psychotropic medications during their continuation course of ECT. Patients who completed the continuation ECT protocol were significantly (p = 0.04) less likely to experience a relapse of depressive symptoms necessitating hospitalization during a 5-month follow-up period than were those who failed to follow the protocol. Indications for use of continuation ECT are discussed and areas for future research are identified. PMID- 11941032 TI - Illness Duration and Acute Response in Major Depression. AB - The authors looked at the association of illness duration and acute treatment response in 1,087 patients with unipolar major depression. Three treatment groups were compared, including a group that received electroconvulsive therapy (ECT), a group that received antidepressants, and a group receiving neither ECT nor antidepressants. Up to an illness duration of 2 years, treatment was significantly associated with recovery; ECT produced significantly better results than other treatments. However, when the illness had lasted >2 years, the ECT treated group showed a significant decline in recovery, and ECT was no longer superior to other treatments. We conclude that duration of illness at index hospitalization has an important influence on treatment response rates and the likelihood of recovery by hospital discharge. PMID- 11941033 TI - Does Fluoxetine Prolong Electrically Induced Seizures? AB - The product literature for the novel antidepressant fluoxetine states that prolonged seizures may occur in patients receiving electroconvulsive therapy (ECT). We compared the seizure durations and incidence of prolonged seizures in 12 consecutive patients receiving ECT plus fluoxetine (20-140 mg/day) with 25 controls receiving ECT in the absence of fluoxetine. No differences were found between the groups. Indeed, within the fluoxetine group, higher doses were associated with slightly shorter average seizure durations, and greater stimulus energy was required to elicit a seizure. We find no evidence that fluoxetine prolongs electrically induced seizures. PMID- 11941034 TI - Protective Effects of Intramuscular Glycopyrrolate on Cardiac Conduction During ECT. AB - The immediate effects of electroconvulsive therapy (ECT) on cardiac conduction, with or without anticholinergic (glycopyrrolate) premedication, were systematically assessed in 19 patients. The resumption of cardiac rhythm after ECT was significantly delayed in treatments without glycopyrrolate. There was no apparent clinical impact of this phenomenon, even though some patients showed asystole of up to 6 s during nonglycopyrrolate treatments. PMID- 11941036 TI - ECT: Twice or Thrice a Week? PMID- 11941035 TI - Electroconvulsive Therapy in the Treatment of Episodic Aggressive Dyscontrol in Psychotic Patients. AB - Five patients with chronic psychosis and episodic aggressive dyscontrol were treated with electroconvulsive therapy (ECT). Four patients also demonstrated clinical evidence of seizure disorder. ECT resulted in marked reduction of both episodic aggressive dyscontrol and clinical seizures, with modest improvement of psychosis. No patient developed clinical signs of organic brain syndrome during ECT. Albeit in a small number of patients, our findings indicate that ECT may have short-term therapeutic effects on episodic aggressive dyscontrol in patients with chronic psychoses. PMID- 11941037 TI - In Reply. PMID- 11941038 TI - Malpractice Litigation and ECT. PMID- 11941039 TI - Fluoxetine and ECT. PMID- 11941040 TI - The Fall and Rise of ECT Within A State Hospital. PMID- 11941041 TI - Is Catatonia a Primary Indication for ECT? PMID- 11941042 TI - Effects of Electroconvulsive Therapy on the Electrocardiogram in Geriatric Patients with Stable Cardiovascular Diseases. AB - Eight elderly patients (mean age +/- SD, 73.2 +/- 9.5 years) were studied with Holter electrocardiographic monitoring for 24 h before and 24 h after an electroconvulsive therapy (ECT) treatment. Patients with a history of cardiac disease but who were not experiencing any active cardiac symptoms were chosen for study. They were not receiving either cardioactive medications or psychotropic agents that can effect cardiac rhythm or conduction. Each patient served as his own control, with direct comparison of the periods before and after the treatment. There were no differences before and after ECT in the electrocardiogram. One minute after the seizure, patients were hypertensive and tachycardic. ECT does not appear to pose additional risk in stable, elderly patients with a history of cardiac disease. PMID- 11941043 TI - Dexamethasone Suppression Tests in the Context of a Double-Blind Trial of Electroconvulsive Therapy and Simulated ECT. AB - Dexamethasone suppression tests (DST) were performed before and after trial treatment in the Leicestershire trial of electroconvulsive therapy (ECT) in practice. Twenty-six depressed patients were studied before treatment, and in 18, tests were repeated immediately after the completion of trial treatments. No consistent pattern of relationship was observed between DST results and response to real or simulated ECT, although the pretreatment relationship between DST results and clinical picture was in accord with the expectations of the literature. PMID- 11941044 TI - Postictal Hyperactive Delirium in ECT: Management with Midazolam. AB - Twenty-one patients who developed one or more episodes of postictal hyperactive delirium (PHD) during the course of electroconvulsive therapy were given a new benzodiazepine drug, midazolam (MDZ), for sedation in an open, uncontrolled clinical trial. MDZ was found to be safe in all patients and was effective as treatment or prophylaxis for PHD in at least 20 patients. Rapid onset of action, brief duration of clinical effects, rapid biotransformation and elimination, wide choice of administration route, and a favorable toxicity profile make MDZ an attractive agent in the management of PHD. The relevant clinical pharmacology of MDZ is reviewed and additional observations regarding PHD are discussed. PMID- 11941045 TI - A Survey of ECT Use in Asia. AB - A questionnaire was mailed to 100 hospitals involved with medical education in 13 Asian countries. Replies were received from 36. The authors compare electroconvulsive therapy (ECT) usage in Asian countries with that in the United States. Marked similarities occur in response rate, number of treatments, and treatment techniques. Differences in anesthesia are associated with a seemingly higher incidence of injury. Bilateral electrode placement and maintenance ECT are widely used. PMID- 11941046 TI - Medical Students and Electroconvulsive Therapy: Their Knowledge and Attitudes. AB - Two consecutive cohorts of medical students who received their fourth year psychiatry training at Manchester University completed knowledge and attitude questionnaires on ECT at the start and conclusion of their clerkships. Their understanding and knowledge of ECT was shown to be greater at the end of the clerkship and their attitudes towards treatment were more positive. The second cohort received an extra teaching session on ECT but this did not confer any additional benefit. It is concluded that increased contact with and knowledge about the treatment has a positive effect on medical students' attitudes. There is still room to improve teaching about ECT. PMID- 11941047 TI - Electroconvulsive Therapy and Chronic Subdural Hematoma. AB - An elderly female patient with major depression was found to have an asymptomatic chronic subdural hematoma. Electroconvulsive therapy (ECT) resulted in full remission of her depression without neurological deterioration. The use of ECT in patients with cerebral lesions is discussed. PMID- 11941048 TI - Clinical Review of Medcraft Corporation's New Brief-Pulse ECT Device. AB - Recently, Medcraft Corporation, a well-known U.S. manufacturer of electroconvulsive therapy (ECT) equipment, introduced a new bidirectional brief pulse model. This machine offers some features that are not currently available on most other ECT devices. In particular, the clinician determines how much stimulus energy the patient receives by setting joules, leading the duration of the electrical stimulation to vary up to 6 s, depending on the patient's dynamic impedance, and the set energy and current levels. The machine is battery powered, completely isolating it from wall current. We summarize our experience with this device, reviewing its features, safety, practicality, and cost. PMID- 11941049 TI - Termination of ECT-Induced Prolonged Seizures. PMID- 11941050 TI - Abdominal Aortic Aneurysm and ECT Administration. PMID- 11941051 TI - Insurance Surcharges for ECT. PMID- 11941052 TI - The 1978 and 1990 APA Task Force Reports. PMID- 11941053 TI - A Retrospective Study of Maintenance Electroconvulsive Therapy in a University Based Psychiatric Practice. AB - We reviewed the log book of our university-based electroconvulsive therapy (ECT) service for the years 1981 to 1987. We identified 10 patients treated with monthly maintenance electroconvulsive therapy (ECT-M). These 10 patients received 3% of the ECT treatments given in this 7 year period. The review of their charts suggests that ECT-M is generally reserved for patients who are older (i.e., over 60 years of age), suffering from delusional depression and/or depressive pseudodementia, and have a history of poor response or tolerance to medications but good response to ECT. The patients had fewer hospitalizations in the 18 months after initiating ECT-M than during the 18 months preceding ECT-M (mean of 3.1 vs. 0.3, respectively, p < 0.001). However, this apparent efficacy of ECT-M may be confounded by the concurrent use of medication. A review of the literature reveals only descriptive studies on ECT-M and shows that our data are congruent with these published studies. The relative value of maintenance ECT and its specificity remain unknown. Its apparent impact on hospitalization rates and safety warrant controlled trials. PMID- 11941054 TI - Full and Abbreviated Courses of Maintenance Electroconvulsive Therapy. AB - The authors describe their experience with maintenance electroconvulsive therapy administered to 10 patients, using an abbreviated or a full maintenance schedule. Recommendation for either form of treatment was made on clinical grounds. Patients with major depressive episodes with delusional features appear to respond best to maintenance ECT. PMID- 11941055 TI - Concurrent and Close Temporal Administration of Lithium and ECT. AB - The clinical effects of concurrent and close temporal administration of electroconvulsive therapy (ECT) and lithium (Li) were investigated by chart review. Three groups of patients were compared on the basis of ECT-related complications, total length of hospital stay (LOS), and post-ECT hospital stay. The groups consisted of patients administered concurrent ECT and Li (n = 27); patients administered Li within 24 h prior to ECT or within 48 h post-ECT (n = 49); and patients administered ECT without concurrent or close temporal administration of Li (n = 100). Prolonged or severe post-ECT confusion was significantly associated with close timing of administration of Li relative to the ECT course. The groups did not differ in the number of complications or the total LOS, although the post-ECT LOS was significantly longer in the group administered Li in close temporal association with ECT. These data suggest that caution should be exercised in the combined use of these treatment modalities. PMID- 11941056 TI - Modification by Chronic Antidepressant Treatment of the Effect of Electroconvulsive Shock on Body Temperature in Mice. AB - The time course of rectal temperature following a single electroconvulsive shock (ECS) was measured in mice 24 h after a single i.p. administration (10 mg/kg) of desipramine, fluoxetine, amitriptyline, pargyline, or saline, as well as 24 h, 1 week, and 2 weeks after the last of 14 once-daily administrations of these antidepressants. There was a decrease of >2 degrees C following a single ECS lasting for at least 90 min, which was not altered by single-dose antidepressant treatment. Following chronic antidepressant treatment, there was a marked attenuation 24 h after the last dose of all drugs used. There was no attenuation 1 and 2 weeks following chronic treatment. Taken together with earlier studies, ECS affects body temperature similarly to 8-OH-DPAT [8-hydroxy-2-(di-n propylamino)-tetralin] and clonidine. However, the mechanism is not yet clear. PMID- 11941057 TI - Effect of Monoamine Oxidase Inhibition, Thiopental Anesthesia, and Electroconvulsive Shock (ECS). AB - A series of experiments were conducted to investigate the effect of the monoamine oxidase inhibitor (MAOI), phenelzine, on thiopental anesthesia, electroshock, and blood pressure. After demonstrating that phenelzine (1 mg/kg/day orally) produced the same decrease in pig platelet monoamine oxidase as seen in humans, we compared MAOI-treated vs. non-MAOI-treated groups of pigs following anesthesia and electroconvulsive shock (ECS). In the majority of pigs, the increase in blood pressure associated with general anesthesia and ECS is no greater after MAOI treatment than after no MAOI treatment. One phenelzine-treated pig demonstrated an excessive catecholamine surge and elevated blood pressure. PMID- 11941058 TI - Hemodynamic Responses to ECT After Bilateral Adrenalectomy. AB - The case of an elderly woman who underwent successful electroconvulsive therapy (ECT) following bilateral adrenalectomy is presented. In contrast to previous animal and human studies, sharp rises in heart rate and blood pressure were observed immediately post seizure, despite pretreatment with nifedipine. These findings indicate that the hemodynamic changes typical of ECT are not necessarily dependent upon increases in circulating epinephrine, and they confirm previous assertions that the therapeutic effect of ECT is not dependent upon intact adrenal function. PMID- 11941059 TI - ECT in an Elderly Patient with Skull Defects and Shrapnel. AB - Electroconvulsive therapy (ECT) was successfully used to treat a patient with large neurosurgical defects of the cranium and small pieces of shrapnel in the zygoma and neck. Other patients with a variety of skull defects have also been treated successfully. Theoretical considerations of current pathway in ECT would suggest an alteration through the brain when the skull is deformed, but this does not appear to affect clinical outcome. PMID- 11941060 TI - ECT for Depression in the Presence of Myasthenia Gravis. AB - A 68-year-old woman with myasthenia gravis of 47 years duration was treated with ECT for an episode of depression. No complications with anesthesia or the ECT procedure were encountered. The depressive symptoms resolved as expected. The safety of ECT in myasthenia gravis is emphasized. PMID- 11941061 TI - ECT in Secondary Mania, Pregnancy, and Sickle Cell Anemia. AB - While ECT has been established as a safe treatment during pregnancy, this case report finds it useful in a patient with sickle cell anemia and mania. Treatment was uneventful and relieved mania occurring secondary to a sickle cell crisis. PMID- 11941062 TI - Repeated ECT Stimuli and the Seizure Threshold. PMID- 11941063 TI - A Reply to Swartz: Abortive Seizures and Subconvulsive Stimuli Are Apples and Oranges. PMID- 11941065 TI - Measurement of Seizure Activity in ECT Research. PMID- 11941064 TI - A Reply to Sackeim and Devanand: Edible Conflicts. PMID- 11941066 TI - ECT in Modern Literature. PMID- 11941067 TI - The Effect of Electroconvulsive Therapy on Tardive Dyskinesia: A Prospective Study. AB - In a prospective study of the effect of electroconvulsive therapy (ECT) on tardive dyskinesia, one patient (of nine) showed dramatic improvement following her fourth treatment. While ECT was effective in only one patient, further studies are warranted to elucidate the mechanism of action of ECT on this movement disorder. PMID- 11941068 TI - Repeated Electroconvulsive Shock Selectively Alters gamma-Aminobutyric Acid Levels in the Rat Brain: Effect of Electrode Placement. AB - The neurochemical mechanisms of electroconvulsive therapy (ECT) are not fully elucidated. We examined the effects of electroconvulsive shock (ECS) on brain gamma-aminobutyric acid (GABA) systems. Male Wistar-Furth rats were given ECS via auricular (ear-clip) or corneal electrodes once per day (120 V, 0.5 s) for 10 consecutive days. Two groups of sham ECS rats, one for auricular placement and one for corneal placement, served as controls. Current was measured and seizures were scored during each ECS trial. Rats receiving ECS via corneal electrodes were subjected to more electrical current compared to rats treated with auricular electrodes. Although both groups exhibited behavioral seizures of similar duration, electrode placement had a differential influence on the expression of tonic hindlimb extension and clonic hindlimb activity over the 10-day regimen. GABA levels were increased in all brain regions examined in rats treated with auricular electrodes except the hippocampus and nucleus accumbens; rats treated with corneal electrodes exhibited GABA increases in the hippocampus, frontal cortex, hypothalamus, and olfactory bulbs; a significant decrease in nucleus accumbens; and no change in the substantia nigra and striatum. The mode of ECS delivery selectively alters the pattern of regional alterations of brain GABA level induced by ECS. This effect may be a function of current intensity or localization. PMID- 11941069 TI - Intraocular Pressure Changes in Nonglaucomatous Patients Undergoing Electroconvulsive Therapy. AB - Before, during, and after electroconvulsive therapy (ECT), intraocular pressure (IOP) was evaluated in ten patients with a negative history for ocular disease. IOP values were obtained using an applanation tonometer. IOP increased dramatically, returning to baseline within 90 s after completion of seizure activity. We conclude that the magnitudes of the IOP elevation, although not hazardous in nonglaucomatous eyes, could possibly compromise ocular functioning in patients with severe glaucoma despite the transient nature of this relative increase in IOP. PMID- 11941071 TI - New Data on the Methohexital-Thiopental-Arrhythmia Issue. AB - Previous reports of a much greater incidence of arrhythmias with thiopental anesthesia for electroconvulsive therapy (ECT) compared to methohexital were reexamined and the possible role of hypercapnia reconsidered. Review of 50 treatments with each agent in 13 patients who had received each at our facility revealed only one brief episode of arrhythmia. The importance of postictal hyperventilation in preventing hypercapnia and associated arrhythmias was suggested. PMID- 11941070 TI - Thyrotropin and Prolactin Secretion During ECT: Implications for the Mechanism of ECT Action. AB - The effect of electroconvulsive therapy (ECT) on serum thyrotropin (TSH) and prolactin (PRL) was systematically investigated in two studies: a simulated ECT (SECT) versus real ECT controlled design, and one in which a more sensitive TSH assay was used. In addition to the expected PRL response, a small but consistent TSH increase was demonstrated, especially when the new TSH assay was used. The implications of these findings concerning the underlying ECT mechanisms mediating these effects are discussed and the involvement of thyrotropin-releasing hormone (TRH) and serotonin is stressed. PMID- 11941072 TI - Depression, Eye Blink Rate, Psychomotor Retardation, and Electroconvulsive Therapy-Enhanced Dopamine Receptor Sensitivity. AB - The effect of electroconvulsive therapy (ECT) and antidepressant drugs on eye blink rates and on a measure of psychomotor retardation was compared in two samples of neuroleptic-free subjects suffering from DSM III major depression and matched on admission by Hamilton and psychomotor retardation scores. Both treatments caused a significant change in eye blink and psychomotor retardation measures. There was no significant difference between treatment groups, although the ECT-treated subjects included those with the largest eye-blink increases. There was no correlation between Hamilton scores and eye blink rates or psychomotor retardation scores for any of the groups or stages of treatment. PMID- 11941073 TI - Outpatient Electroconvulsive Therapy: Efficacy and Safety. AB - Thirty-two outpatients with a mean age of 68 years were observed in a naturalistic study of outpatient electroconvulsive therapy (ECT) for the management of recurrent mood disorder. Sixty-nine percent of the patients responded favorably to ECT and were either discharged well from therapy or continue in treatment. A rehospitalization rate of 9% supports the efficacy of this treatment. We find outpatient ECT safe for the long-term management of recurrent depressive illness in elderly patients. The low dropout rate demonstrates that patient acceptance is equal to that of other outpatient treatments. PMID- 11941074 TI - Electroconvulsive Therapy of the Lethal Catatonia Syndrome. AB - Lethal catatonia (LC) is a life-threatening syndrome associated with diverse neuropsychiatric or systemic disorders. Neuroleptic agents appear inadequate in treating LC. We report a case of LC successfully treated by electroconvulsive therapy (ECT) that adds to the experience of ECT as a safe and effective treatment for LC occurring in the context of the major psychoses. Anecdotal evidence suggests that ECT is dramatically effective in LC regardless of etiology. The use of ECT in the treatment of neuroleptic malignant syndrome (NMS), viewed here as a subtype of LC, is considered and compared with that of specific drug therapies for NMS. PMID- 11941075 TI - Detection of Prolonged Seizure by Audible EEG. PMID- 11941076 TI - Electroconvulsive Therapy and Fluoxetine. PMID- 11941077 TI - Neuroleptic Malignant Syndrome and Electroconvulsive Therapy. PMID- 11941078 TI - Electrode Placement: A Clinician's Guide. PMID- 11941079 TI - Biogenic Amine Metabolites During Electroconvulsive Therapy of Melancholic Patients. AB - We assayed the urinary neurotransmitter metabolites 3-methoxy-4 hydroxyphenylglycol (MHPG), homovanillic acid (HVA), and 5-hydroxy-indoleacetic acid (5-HIAA) in unipolar depressed patients before and after a simulated electroconvulsive therapy ECT (SECT), and during course of 10 ECT sessions. A repeated measures analysis of variance (ANOVA) showed no significant changes in the three-metabolite excretion during the course of ECT. Planned comparisons performed after ANOVA revealed a trend for HVA and 5-HIAA levels to increase after SECT and a significantly higher MHPG excretion after the 10th ECT session. Seven depressed patients who responded favorably to ECT (reduction in Hamilton Rating Scale for Depression score of 50% or more) but not the seven nonresponders had significantly higher MHPG excretion after the final ECT compared to baseline levels. A significant relationship was found between low pretreatment MHPG excretion and therapeutic response. PMID- 11941080 TI - Rat Brain Concentration of Fluphenazine During a Course of Electroconvulsive Shock. AB - The brain concentration of fluphenazine was measured in two groups of rats, one treated with fluphenazine and electroconvulsive shock, and the other with fluphenazine only. No difference was found in brain fluphenazine concentration between the two groups. The experiment was replicated with a larger number of rats and the same result was obtained. Electroconvulsive shock does not increase the concentration of fluphenazine in brain tissue. PMID- 11941081 TI - Maintenance Electroconvulsive Therapy in Clinical Practice. AB - Five cases of maintenance electroconvulsive therapy (ECT) are reviewed to illustrate the indications for this treatment. The patients' conditions described are consistent with the recommendations of the 1990 American Psychiatric Association Task Force on ECT. Guidelines and examples for selecting patients for maintenance ECT are presented, but when to preferentially treat with continuation pharmacotherapy versus ECT requires further study. Optimal dosing, interval between treatments, and duration of treatment remain questions. The logistics of the treatment are briefly presented and should be easily incorporated into most ECT programs. PMID- 11941082 TI - The Use of Metrazol in the Treatment of Patients with Mental Diseases. PMID- 11941083 TI - Prospective Electroconvulsive Therapy in the Presence of Intracranial Tumor. AB - Three cases of patients known to have cerebral meningiomas at the time of electroconvulsive therapy (ECT) are reported. They represent the seventh to ninth reported cases of patients with intracranial neoplasms treated prospectively with ECT. No patient suffered adverse effects referable to the presence of the tumors, and each obtained a good therapeutic response. The English language literature concerning the use of ECT in patients with intracranial neoplasms is reviewed, and it is concluded that ECT may be safely administered prospectively to patients with intracranial meningiomas after the risk of neurological complications has been evaluated with the appropriate diagnostic studies. PMID- 11941084 TI - Caffeine-Associated Cardiac Dysrhythmia During ECT: Report of Three Cases. AB - Caffeine is increasingly being used by ECT practitioners to augment seizure duration in selected patients. We note one report of adverse cardiovascular effects with this procedure. We report three cases of severe cardiac dysrhythmia, all appearing in the present of caffeine. The three patients were elderly, with a prior history of cardiac illness. The authors recommend the use of nonpharmacologic alternatives for the treatment of brief seizures, and careful review of each patient's cardiac history before utilizing caffeine. They further urge continued monitoring in those patients for whom caffeine is introduced. Further prospective studies are needed to assess carefully the risk of caffeine pretreatment in patients with cardiac disease. PMID- 11941085 TI - Ease of Use of Disposable Mouthguards During ECT. AB - Disposable, molding-type soft plastic mouthguards often used in contact sports are recommended for use in electroconvulsive therapy (ECT). They are easy to use, encourage good hygiene, and prevent soft tissue and tooth injuries during ECT. PMID- 11941086 TI - Frequency of ECT Treatments. PMID- 11941087 TI - Further Characterization of Mania as a Side Effect of ECT. PMID- 11941088 TI - Choice of Stimulus Electrode Placement: Clarification by the APA Task Force on ECT. PMID- 11941089 TI - The FDA Proposal to Reclassify ECT Devices. PMID- 11941090 TI - Effects of Labetalol on Hemodynamics and Seizure Duration During ECT. AB - Beta blockers are commonly used to attenuate the transient increase in heart rate (HR) and blood pressure (BP) that accompany electroconvulsive therapy (ECT). Recent reports have suggested that several beta blockers have anticonvulsant properties and hence could be countertherapeutic with ECT. In a randomized, double-blind controlled study of the dose response effects of intravenous labetalol on ECT hemodynamics and seizure duration, labetalol exhibited dose dependent reduction in HR and rate pressure product (RPP) at the 5 and 10 mg doses compared to placebo. However, no significant differences between labetalol 5 mg, 10 mg, or placebo were found on BP or seizure duration. PMID- 11941091 TI - Electroconvulsive Therapy and Brain Glucose Metabolism. AB - The local cerebral metabolic rate for glucose (LCMRGlc) was evaluated in four patients before undergoing a course of electroconvulsive therapy (ECT) and after its completion. There was no reduction in LCMRGlc when studied 1 day after the last course of ECT. There was, however, the suggestion that metablic rates may increase in the middle frontal gyrus after significant time has elapsed since the last course of ECT. PMID- 11941092 TI - Do Opioid, alpha(2)-Adrenoreceptors, or Benzodiazepine Receptors Mediate ECS Induced Hypothermia in Mice? AB - A single electroconvulsive shock in TO mice produced a drop in body temperature of 1.5-2 degrees C, which was maintained for up to 60 min. This hypothermic response was not attenuated by the opioid antagonist i.p. naltrexone (0.3, 1 mg/kg); the alpha(2)-adrenoreceptor antagonist i.p. RX811059 (0.05, 0.5 mg/kg), or the benzodiazepine receptor antagonist i.p. Ro 15-1788 (10, 20 mg/kg). The doses of antagonist used in this study had no effects on body temperature by themselves but significantly attenuated the hypothermia induced by their respective agonists, confirming their receptor-blocking activity. These findings suggest that electroconvulsive shock-induced hypothermia is not mediated by the release of either noradrenaline or by an endogenous ligand acting at opioid or benzodiazepine receptors. PMID- 11941093 TI - Hypertension and Tachycardia During Electroconvulsive Therapy: To Treat or Not to Treat? AB - ECT is associated with hypertension and tachycardia for a brief period of time. In some patients, as those with severe cardiovascular disease, reduced intracranial compliance, and vascular pathology that may precipitate hemorrhage, it is reasonable to modify the cardiovascular response. Various techniques are reviewed and a method of control in use in our practice is recommended for its efficacy and safety. PMID- 11941094 TI - Clozapine, ECT, and Schizoaffective Disorder, Bipolar Type. AB - Opinions differ regarding the risks and benefits of the concurrent use of antipsychotic medication and ECT. A case example is presented of the safe and effective concurrent use of ECT and the newly available neuroleptic clozapine in a young patient with schizoaffective disorder, bipolar type. The patient's pulse did climb as high as 170-180 beats/min during a few of the seizures. More experience is needed with the combined use of clozapine and ECT. PMID- 11941095 TI - Post-ECT Dyskinesia. AB - Three depressed individuals who developed a transient dyskinesia after right unilateral, nondominant electroconvulsive therapy (ECT) are described. The patients responded well to ECT in terms of their affective disorders, and were discharged free of permanent ECT-induced movement disorder symptoms. Risk factors and possible neurochemical changes underlying this phenomenon are discussed. PMID- 11941096 TI - Electroconvulsive Therapy and Friedreich's Ataxia. AB - A woman with Friedreich's ataxia, a hereditary neurodegenerative disorder affecting primarily the spinal cord and cerebellum, had intractable depression in the context of bipolar disorder. Electroconvulsive therapy resulted in substantial improvement of depressive symptoms while not adversely affecting the patient's neurologic status. PMID- 11941097 TI - Electroconvulsive Therapy in Paget's Disease and Hydrocephalus. AB - Electroconvulsive therapy (ECT) was successfully used to treat depressive illness in a patient with Paget's disease of the skull and hydrocephalus with a functioning shunt. PMID- 11941098 TI - ECT in the Presence of Severe Cervical Spine Disease. AB - We present a case of the safe administration of electroconvulsive therapy (ECT) to a depressed patient with severe, unstable cervical spine disease. Using adequate muscle relaxation, a course of ECT treatments was given with excellent results. Although fears about the use of ECT in the presence of spine disease persist, attention to modern ECT technique allows a safe treatment course. PMID- 11941099 TI - Bladder Rupture During ECT. AB - We report a case of intraperitoneal bladder rupture during electroconvulsivee therapy. We consider that the likely cause of this complication was powerful abdominal muscle contraction, during an unmodified seizure, on a bladder distended by failure to void urine before treatment and the anticholinergic effects of medication. The literature is reviewed, a similar case is discussed, and we make recommendations for the prevention of this complication. PMID- 11941100 TI - Propranolol in ECT. PMID- 11941101 TI - Electrode Placement in Maintenance Electroconvulsive Therapy. PMID- 11941102 TI - Quantity of Prolactin Released by ECT Seizure. PMID- 11941103 TI - Who Guards the Mouthguards? PMID- 11941104 TI - More on Mouthguards. PMID- 11941105 TI - Pharmacotherapy and ECT. PMID- 11941106 TI - Prediction of ECT Response in Melancholia. AB - None of 15 individual Hamilton depression scale items obtained at baseline, or five depression factors derived therefrom, significantly predicted an adjusted posttreatment Hamilton depression score after six unilateral or bilateral ECTs in a sample of 47 melancholic men. Excluding patients with nonmelancholic syndromes from ECT samples attenuates the previously reported predictive value of individual or grouped clinical psychopathological features of depression. PMID- 11941107 TI - Electroconvulsive Therapy-Induced Prolactin Release as an Epiphenomenon. AB - Electroconvulsive therapy (ECT)-induced prolactin release has been described by many investigators with the expectation that it reflects brain chemistry and thereby the neurochemical pathology of depression or the therapeutic impact of the ECT seizure, analogous to a blood drug level. Neuroendocrine evidence is reviewed to construct plausible mechanisms for ECT-induced prolactin release dependent on three sequential steps outside the brain: posterior pituitary change, transport to anterior pituitary, and prolactin release from anterior pituitary. These mechanisms also explain the prolactin elevations induced by suckling and by estradiol exposure. One mechanism posits seizure-induced interruption of the dopamine flux from posterior to anterior pituitary and consequent decrease of tonic inhibition on prolactin release. The other mechanism depends on a short-chain peptide prolactin-releasing factor. The substantial likelihood of strong posterior pituitary control of ECT-induced prolactin release suggests that it reflects brain chemistry only dimly. PMID- 11941108 TI - Atropine and Cognitive Performance After Electroconvulsive Therapy. AB - Two groups of patients receiving bilateral, moderately suprathreshold electroconvulsive therapy (ECT) were compared in their cognitive functions after receiving either 0.5 mg atropine i.v. or no atropine before ECT. The patients were tested for cognitive functions after four treatments using various measures, including orientation; retrograde (verbal and visuospatial) memory for information acquired about 15 minutes pretreatment and tested for about 1 hour posttreatment; anterograde (verbal and visuospatial) memory for information acquired about 1 hour, 30 minutes posttreatment and tested for both immediately after learning and 20 minutes later; and retrieval from semantic memory as assessed by word fluency. We found no effect of 0.5 mg atropine on cognitive performance, even though anticholinergic drugs that cross the blood brain barrier might be expected to affect cognition after ECT. PMID- 11941110 TI - Electroconvulsive Therapy in the Treatment of the Neuroleptic Malignant Syndrome. AB - In a review of 734 published cases of neuroleptic malignant syndrome (NMS), we found 665 with sufficient data to allow statistical analyses. Forty-eight received ECT either during or shortly after an episode of NMS and were compared to control cases who received no specific treatment for their episode. We also compared the mortality rates in these two groups to data previously analyzed for specific drug treatments (i.e., amantadine, bromocriptine, L-dopa, and dantrolene). We find a substantially better outcome in the specific drug-treated (mortality rate of 9.7%) or ECT-treated (mortality rate of 10.3%) groups compared to the group receiving no specific treatment (mortality rate of 21%). We conclude that ECT is safe to use shortly after an episode of acute NMS has resolved and, furthermore, that ECT is probably safe in the actual treatment of NMS provided concomitant neuroleptics are discontinued. PMID- 11941109 TI - Disorientation and Bilateral Moderately Suprathreshold Titrated ECT. AB - Thirty-seven inpatients with major depression were assessed for postictal and interictal disorientation after they received 8 of 12 ECTs. In 20 patients, four of the eight assessments were after simulated ECT only. Only real, but not simulated, ECT produced postictal disorientation. Postictal disorientation was greatest after the first treatment, less after the second, and did not change in later assessments. It was shortest for person, longer for place, and longest for time, and showed a temporal time gradient. Interictal disorientation increased with the number of treatments. Two electrical stimulus variables (seizure duration and electrical stimulus intensity) correlated with the length of postictal disorientation. The influence of seizure duration and stimulus variables were independent of each other. The influence of the electrical stimulus variables was independent of the influence of demographic variables. These, however, did affect the length of postictal disorientation. PMID- 11941111 TI - Severe Medical and Neurologic Complications Associated with Near-Lethal Catatonia Treated with Electroconvulsive Therapy. AB - In a patient with near-lethal catatonia, complications progressed from mild fever and tachycardia to severe hyperthermia, malignant hypertension, and cortical and subcortical dysfunction with increased urine cortisol and catecholamine concentrations, bilateral extensor toe reflexes, decerebrate posturing, and Cheyne-Stokes respirations. Electroconvulsive treatment reversed these changes and prevented almost certain death. PMID- 11941112 TI - Normalization of Augmented TRH Stimulation Test in Response to ECT. AB - A 78-year-old woman with a major depression and subclinical hypothyroidism was successfully treated with ECT. Her abnormally augmented pretreatment thyrotropin releasing hormone (TRH) stimulation test normalized. We discuss the interaction of electroconvulsive therapy (ECT) and the thyroid axis. PMID- 11941113 TI - Atypical Cholinesterase and Prolonged Apnea During Electroconvulsive Therapy. AB - A 62-year-old man developed prolonged apnea during ECT with succinylcholine. He was found to have atypical cholinesterase. Subsequent ECTs were given using atracurium as the muscle relaxant. A review of the finding of atypical cholinesterase is presented along with a discussion of the use of atracurium. PMID- 11941114 TI - Electroconvulsive Therapy in a Depressed Patient with an Intracranial Arachnoid Cyst: A Brain Magnetic Resonance Imaging Study. AB - We describe a depressed man with an asymptomatic arachnoid cyst of the left sylvian fissure who was successfully treated with ECT. Magnetic resonance imaging of the brain before and after the course of therapy revealed no change in the cyst nor any injury to the surrounding brain parenchyma. PMID- 11941115 TI - Atrial Fibrillation, Cardioversion, and Electroconvulsive Therapy. AB - An elderly female patient with major depression and incidental atrial fibrillation received electroconvulsive therapy (ECT). Minutes after two different treatments on two separate occasions, she converted to a normal sinus rhythm. Although atrial fibrillation itself is associated with an increased risk of thromboembolic stroke, cardioversion without anticoagulation in individuals with nonacute atrial fibrillation is associated with a 1-2% incidence of embolization. Cardioversion during ECT may be infrequent but its possibility should alert clinicians to the consideration of prophylactic anticoagulation for some patients undergoing ECT. PMID- 11941117 TI - A Scientific Society for ECT. PMID- 11941116 TI - Fluoxetine and Prolonged Seizure. PMID- 11941118 TI - ECT Treatment of Demented Elderly Patients with Major Depression: A Retrospective Study of Efficacy and Safety. AB - Retrospective analysis of a 4-year sample of demented patients who had major depression and who were treated with electroconvulsive therapy (ECT) showed that they responded as well to ECT as did a previously studied group of elderly depressed subjects from our institution. Cardiac side effects were no greater in this group, but some patients did have transitory increased confusion. ECT is a safe and effective therapy for the demented patient who has concomitant major depression and who has proven to be resistant to tricyclic antidepressant drug therapy, is intolerant of such medicines, or requires emergency treatment. PMID- 11941119 TI - The Reliability of Computer-Processed EEG in the Determination of ECT Seizure Duration. AB - The reliability of single-channel analog EEG and two-channel, computer-processed EEG (cEEG) in determining seizure duration during electroconvulsive therapy (ECT) was studied in 144 consecutive individual treatment sessions of 14 hospitalized patients. Seizure durations determined by post hoc, blind readings of data generated by each method were compared. These estimates of seizure duration were also compared to those determined by the "cuff" method. We found that under our study conditions, the cEEG method was more reliable than the EEG method in two tests of reliability: the cEEG method had fewer readings with a discrepancy of 10 s or more between readers, and had a higher degree of correlation between readers. There was no difference, however, between the cEEG and EEG methods when comparing mean differences and the mean absolute difference between readers within the methods. We also found that the EEG and cEEG methods detected seizure durations that were substantially longer than those detected by the "cuff" method. We conclude that under our study conditions, cEEG was more reliable than analog EEG in certain measures of reliability. PMID- 11941120 TI - Patient and Family Perspectives of Electroconvulsive Therapy: Correlation with Outcome. AB - We surveyed 25 patients about to undergo electroconvulsive therapy (ECT) and one relative of each patient for their attitudes, knowledge, and opinions about ECT; and then resurveyed them after the course of ECT. Patients were rated with the Hamilton Depression Rating Scale or Young Mania Rating Scale, based on diagnosis, before and after ECT. Patients and family members had highly positive attitudes toward ECT both before and after treatment, and after the course felt strongly that the ECT was beneficial. Baseline depression severity gave a high correlation with positive post-ECT attitudes. PMID- 11941121 TI - Change in Attitude Toward Electroconvulsive Therapy: Effects of Treatment, Time Since Treatment, and Severity of Depression. AB - Attitudes toward electroconvulsive therapy (ECT) of patients with major depressive episodes who are treated with ECT were evaluated before the beginning of treatment, 1 to 2 days after completion of the 12th treatment, and 6 months after the termination of the series using a questionnaire (adapted from Freeman and Kendall, 1980). Attitudes toward ECT become more positive after treatment, and remain so at the 6-month follow-up. Attitude changes correlate with changes in depressive symptoms and with subjective side effects during treatment. Patients who had a prior course of ECT had more knowledge of ECT but not a more positive attitude. PMID- 11941122 TI - Is Unilateral Nondominant ECT as Efficient as Bilateral ECT? A New Look at the Evidence. AB - A survey of 24 comparisons of the antidepressive efficiency of bilateral and unilateral nondominant electroconvulsive therapy (ECT), fulfilling strict methodological criteria, shows a preponderance of studies in favor of bilateral ECT. However, the inferiority of unilateral nondominant ECT can be explained by submaximal seizure activity due to short interelectrode distance, barely suprathreshold stimulation, and combination with benzodiazepine drugs. With optimal technique, unilateral nondominant ECT may be as efficient as bilateral ECT. Furthermore, the cognitive dysfunction is less and of shorter duration. There is no reason to retain or return to bilateral ECT, but instead we should pay attention to technical improvements of unilateral nondominant ECT. PMID- 11941124 TI - Seizure Generalization and Unilateral Electroconvulsive Therapy. AB - The author responds to Ottosson's exposition of the variables that determine the relative efficacies of unilateral and bilateral ECT. To Ottosson's prescription for maximizing the therapeutic impact of unilateral ECT, he adds the administration of a markedly suprathreshold brief pulse stimulus train of.5 to 1.5 ms pulse width and >/=2 s duration. PMID- 11941123 TI - Optimizing Unilateral Electroconvulsive Therapy. AB - Review of the unilateral-bilateral literature cannot identify the factors that optimize the efficacy of right unilateral electroconvulsive therapy (ECT) (RUL), and we should not confuse hunches with facts. With the exception of stimulus dosing, there is little experimental support that the conditions suggested by Dr. J.-O. Ottosson affect RUL efficacy. The theoretical position that eliciting a full, maximally generalized seizure provides the basis for optimal efficacy also lacks experimental support and is imprecise. Further, the conditions proposed to optimize RUL efficacy also augment its side effect profile. We do not know whether an optimal form of RUL will retain cognitive advantages if compared to optimal bilateral ECT. We also lack a clinically useful marker that an optimal form of RUL has been delivered. Under these conditions, it seems appropriate that the choice of electrode placement be made on an informed case-by-case basis. PMID- 11941125 TI - Combined Use of Clozapine and Electroconvulsive Therapy. AB - We used electroconvulsive therapy (ECT) together with clozapine in two patients to treat delusional depression. Both patients had responded minimally to ECT. There were no difficulties in the course of treatment. The combination may be useful in patients with ECT-resistant psychosis. PMID- 11941126 TI - ECT for Refractory Parkinson's Disease. PMID- 11941127 TI - Are ECT Devices Underpowered? PMID- 11941128 TI - Cerebral Edema and Vascular Permeability to Serum Proteins Following Electroconvulsive Shock in Rats. AB - The regional brain specific gravity and the cerebrovascular permeability to serum proteins were investigated in rats subjected to electroconvulsive shock (ECS) with different stimulus intensity and different stimulus periods. The following experimental situations were studied: one ECS daily for 9 days (50 mA in 0.3 s), one ECS daily for 9 days (50 mA in 0.9 s), and one ECS three times weekly for 4 weeks (50 mA in 0.3 s). Age-matched animals receiving sham ECS served as controls. In the group having stronger stimulus intensity and in the one treated for 4 weeks, there was an increase in tissue water content in the hypothalamus and in both hypothalamus and hippocampus, respectively. In none of the experimental groups could cerebral edema be demonstrated in parietal cortex or in white matter. The findings point to an increased blood-to-brain transfer of water with increasing stimulus intensity and with the length of the ECS series. The study showed no increased cerebrovascular permeability to serum proteins and no signs of neuronal damage in any of the experimental groups. PMID- 11941129 TI - Factors Influencing Response to Bilateral Electroconvulsive Therapy in Major Depression. AB - The records of 52 patients with major depression who were treated with bilateral electroconvulsive therapy (ECT) were reviewed. Responders and nonresponders were compared on demographic, clinical, and treatment parameters. ECT nonresponders had a longer duration of current depressive episode as well as a greater initial severity of depression. The groups did not differ in age, sex, polarity, presence of psychosis, pre-ECT pharmacotherapy, and treatment parameters other than total electrical charge administered. Patients with long episode duration and greater severity of illness may represent a subgroup of major depressives relatively refractory to ECT and warranting novel therapeutic approaches. PMID- 11941130 TI - Psychiatric Malpractice and ECT: A Review of 1,700 Claims. AB - A total of 1,700 psychiatric malpractice claims filed from 1984 to 1990 were reviewed. Twenty-two cases involved ECT. Only four cases had complaints that referred to side effects, complications, or appropriateness of ECT. In the remainder, ECT was given in the past and/or the complaint was unrelated to the use of ECT. These data show a low frequency of lawsuits related to the use of ECT. The cost of resolving ECT cases was less than the average of all psychiatric claims. Patient satisfaction with the effectiveness of ECT may explain these findings. PMID- 11941131 TI - Survey of Anesthetic Coverage of Electroconvulsive Therapy in the State of Pennsylvania, 1988. AB - To ascertain the current status of anesthesia care for electroconvulsive therapy (ECT) on a statewide basis, 54 facilities identified as providing ECT in the State of Pennsylvania were sent a specially designed 127-item questionnaire. Of 27 (50%) respondents, 25 (93%) included anesthesia personnel (anesthesiologists, nurse anesthetists, and anesthesia residents) on the treatment team and 2 (7%) did not. The questionnaire included items concerning pretreatment evaluation, preparation of patients, anesthesia administration, postanesthetic considerations, treatment locale, and patient profile. The items were derived from the American Psychiatric Association (APA) Task Force Report on Electroconvulsive Therapy, the Guidelines for Patient Care in Anesthesiology of the American Society of Anesthesiologists (ASA), the Standards for Nurse Anesthesia Practice of the American Association of Nurse Anesthetists (AANA), and items that reflected guidelines of the ASA and AANA. The responses of each profession were compared to its own organizational guidelines and to the other guidelines. The professions did not adhere to their own guidelines; the lowest compliance was for psychiatrists to items derived from the APA. Items derived from the APA generally received lower responses from all professions in the study. The study documents the present-day anesthetic coverage of ECT in the State of Pennsylvania, and reflects an almost complete change over a 10-year period (1978-1988) to a two-member team for ECT: a psychiatrist for electroconvulsive treatment and an anesthetist or anesthesiologist for the anesthesia management. PMID- 11941132 TI - Eye Coordination and Vestibular Testing in Electroconvulsive Therapy. AB - Nine depressive patients were examined before bilateral electroconvulsive therapy (ECT), immediately after ECT, and 2 days later with saccadic, pursuit tracking, and Hallpike caloric vestibular tests. Before ECT, all tests showed normal results. Immediately after ECT, the saccadic test displayed dysmetria of the fast eye movements in six of nine patients. The pursuit tracking test showed decreased gain of the slow eye movements with compensatory saccades in seven of nine patients. These findings were for both directions in the horizontal plane. In the caloric vestibular test, the changes were not significant. Two days after ECT, the eye coordination and vestibular testing showed normal function. We conclude that the eye coordination and vestibular function disturbances induced by ECT are transient. PMID- 11941133 TI - Remission of Tardive Dystonia with ECT. AB - A 30-year-old patient with tardive dystonia, who had failed to respond to cessation of neuroleptics, placebo, diazepam, biperiden, propranolol, and clonidine, had an impressive response to courses of electroconvulsive therapy (ECT) on three successive trials. PMID- 11941134 TI - Cyanosis and Dyspnea in Alpha(1)-Antitrypsin Deficiency with ECT. AB - A 40-year-old depressed patient with alpha(1)-antitrypsin deficiency and severe emphysema developed bronchospasm shortly after an initial electroconvulsive therapy (ECT), resulting in discontinuation of treatment. The use of ECT in patients with emphysema and prevention of respiratory complications are discussed. PMID- 11941135 TI - Problems in Maintenance ECT in Bipolar Disorder: Replacement by Lithium and Anticonvulsants. AB - The authors describe three elderly patients whose electroconvulsive therapy (ECT) was characterized by protracted and unsatisfactory courses and frequent relapse. Sustained improvement resulted from the discontinuation of maintenance ECT and continuation of pharmacotherapy with combinations of lithium and carbamazepine or valproic acid. The patients are compared to a more typical patient with bipolar disorder who was successfully treated with long-term maintenance ECT. Difficulties in identifying and treating patients with atypical bipolar disorder are discussed. PMID- 11941136 TI - ECT in Modern Literature (Continued). PMID- 11941137 TI - A Behavioral Approach In Post-ECT Delirium. PMID- 11941138 TI - Labetalol and ECT. PMID- 11941139 TI - Double Stimulation To Elicit an Adequate Treatment. PMID- 11941140 TI - In Reply: Double Stimulation Can Bypass Safety Standards. PMID- 11941141 TI - Follow-Up on Clozapine and ECT. PMID- 11941142 TI - Qualification for ECT. PMID- 11941143 TI - Unmodified Electroconvulsive Therapy of Acute Mania: A Retrospective Naturalistic Study. AB - In this retrospective naturalistic study, clinical records were reviewed to examine the effects of unmodified electroconvulsive therapy (ECT) in 30 manic patients at a psychiatric institute in India. Twenty of the patients were experiencing their first episode of a major mood disorder. ECT was associated with complete remission of mania in 26 (87%) of the patients. The remaining four patients were discharged home with residual hypomanic symptoms. On average, the patients received 5.4 treatments, and the duration from initiation of ECT to discharge from hospital was 12.9 days. The number of treatments was not related to age, age at onset, duration of index episode prior to treatment, or presence of psychosis. No patient developed an organic brain syndrome or sustained a bone fracture during unmodified ECT. One month follow-up data were available in 27 of the 30 patients. Recurrence of manic symptoms following discharge occurred in three (11%) patients. For those in whom 3 month follow-up data were available, all 15 were in complete clinical remission. The induced convulsion, and not repeated administration of general anesthetics, is integral to the antimanic effect of ECT. PMID- 11941144 TI - ECT in a State Hospital Setting. AB - The use of electroconvulsive therapy (ECT) in U.S. state hospitals has markedly decreased in the past 20 years. During this time ECT technique has reached a high level of sophistication. We examine whether ECT still has a place and can be effectively used in a modern state hospital. We conclude that approximately 1% of state hospital patients are candidates for ECT and that patients with major depression, mania, schizoaffective disorder, and schizophrenia may benefit from such treatment. The relative youth and good physical health of state hospital patients referred for ECT reduces the risk of the procedure in this setting. Most of our patients were on civil commitment but retained the capacity to consent to treatment. We discuss the use of inpatient continuation ECT and the treatment of protracted psychotic excitement as issues relevant to state hospitals. PMID- 11941145 TI - Dental Pathology in ECT Patients Prior to Treatment. AB - Dental complications during electroconvulsive therapy (ECT) are uncommon. Nevertheless, dental complications are often cited in ECT-related malpractice litigation. Choice of dental protection during ECT should be based on dental pathology prior to ECT. We describe the results of formal dental examinations of 82 patients prior to ECT and 82 matched psychiatric controls. Dental pathology was frequent in both groups without intergroup differences. The decision to pursue a formal dental consultation prior to ECT should be based upon questions raised on inspection of the oral cavity by the treating psychiatrist. The type of dental protection during ECT should be chosen according to the patient's pre-ECT dental status. PMID- 11941146 TI - ECT in Austria: Is It Still a Valid Treatment? Appraisal of Present Day Standards in a Psychiatric Hospital. AB - We report on the current experience of electroconvulsive therapy (ECT) in a modern regional psychiatric hospital in Austria. The article describes ECT in 120 psychiatric inpatients during 1980-1987. All patients were hospitalized, the main indications for ECT being "neuroleptic treatment resistance" and "catatonia/stupor." Between 1980-1987, an average of 0.7% of admissions (0.6 2.75%) were treated with a course of 10 (62%) or 15 ECTs (25%), with 4% of patients requiring 15 ECTs or more. In more than one-half of the patients, bilateral electrode placement was used. All ECT was done under anesthesia and muscular relaxation. We find ECT to be a valuable treatment-technique for a subgroup of psychotic inpatients. PMID- 11941147 TI - Manic Psychosis and Catatonia Stemming from Systemic Lupus Erythematosus: Response to ECT. AB - A 27-year-old woman with severe psychosis and mania associated with systemic lupus erythematosus was successfully treated with electroconvulsive therapy. Cyclophosphamide was given as an adjuvant. The use of electroconvulsive therapy in patients with lupus and psychiatric disorders is discussed. PMID- 11941148 TI - Acute Administration of Fluoxetine and the Duration of Electrically Induced Seizures. PMID- 11941149 TI - ECT and Grand Mal Epilepsy. PMID- 11941151 TI - Stimulus Dosing Strategies and Unilateral ECT. PMID- 11941150 TI - Recurrent Post-ECT Dyskinesia. PMID- 11941152 TI - In Reply: Stimulus Dosing Strategies and the Efficacy of Unilateral ECT. PMID- 11941153 TI - In Reply to Sackeim et al. PMID- 11941154 TI - ECT and Public Mental Health Services. PMID- 11941155 TI - Safety and Efficacy of ECT in Depressed Patients with Organic Brain Disease: Review of a Clinical Experience. AB - We performed a retrospective review of 5 years of experience with electroconvulsive therapy (ECT) in patients with organic brain lesions. On 26 of 27 occasions, patients with organic brain disease and a concurrent depressive disorder obtained a good affective response to a course of ECT. Patients with diffuse or multifocal brain disease were vulnerable to ECT-induced delirium; however, this was severe enough to compel discontinuation of ECT on only one occasion. We conclude that ECT is an effective treatment for depression in patients with concurrent neurological disease, although patients with degenerative brain disease or diffuse encephalopathy may be especially prone to ECT-induced delirium. PMID- 11941156 TI - Psychiatry Resident and Medical Student Perspectives on ECT: Influence of Exposure and Education. AB - We surveyed medical students doing the required psychiatry clerkship at our hospital and first-year psychiatry residents in our residency training program for their attitudes, knowledge, and opinions about electroconvulsive therapy (ECT) before and after their inpatient rotations. The students' knowledge was lower and their attitudes more negative toward ECT than those of residents at baseline. Upon completion of the rotation, students and residents had more knowledge and highly positive attitudes about ECT and stated that they would undergo ECT if it were recommended. Willingness to undergo ECT if it were recommended correlated with the knowledge measures of ECT. PMID- 11941157 TI - A Comparison Between Etomidate and Methohexital for Anesthesia in ECT. AB - Twenty patients were enrolled in a prospective, randomized, open vial, within patient, crossover design study of methohexital and etomidate anesthesia for electroconvulsive therapy (ECT). Methohexital (1.0 mg/kg) and etomidate (0.3 mg/kg) were each given for two ECTs. While there were no differences in hemodynamics between the etomidate and methohexital groups, etomidate had a 24% longer mean wakeup time than methohexital. There was no difference between etomidate and methohexital in recovery room stay, induction time, or seizure duration. There was no difference in cardiac rhythms. More patients had pain on injection with etomidate than with methohexital. Increased incidence of pain on injection and a longer initial wakeup time are drawbacks to etomidate, but etomidate compared favorably to methohexital in hemodynamics. Etomidate is an acceptable alternative to methohexital, especially when barbiturates may be contraindicated. PMID- 11941159 TI - ECT and Twin Pregnancy. AB - The combination of pregnancy and affective psychosis produces a complex treatment problem. The mother and fetus can experience significant risks from the disorganized behavior associated with mental illness as well as from the medical treatments for the illness. ECT has been recommended as a safe alternative. This article reports on ECT used for a patient pregnant with twins. Multiple gestation compounds the problems encountered with single pregnancies. The report details techniques to monitor fetal well-being. The case illustrates the safe use of ECT when the fetuses and the pregnancy itself are at high risk. PMID- 11941158 TI - The Role of Dopamine in Seizure-Induced Prolactin Release in Humans. AB - Prolactin (PRL) plasma levels rise severalfold following spontaneous or electrically induced seizures. To assess the role of dopamine in this neuroendocrine response, we studied the effect of electroconvulsive therapy (ECT) on plasma PRL after maximal dopamine receptor blockade induced by intravenous metoclopramide. Six patients undergoing ECT received metoclopramide or placebo, in counterbalanced order, 30 min prior to application of the electrical stimulus. Blood samples were obtained at - 15 min, immediately before, and at 15 min intervals for 1 h after the treatment. Under placebo conditions, PRL plasma levels increased nearly sevenfold in response to ECT. The administration of metoclopramide produced a large increase in PRL plasma levels, with no further increase in PRL concentrations after ECT. The data suggest that the PRL plasma rise after ECT involves dopamine as it is not seen when the dopaminergic inhibitory tone, exerted by the hypothalamus on the pituitary lactotroph, has been removed. PMID- 11941161 TI - Combining Clozapine with ECT. AB - We describe the safe and effective use of the combination of clozapine and ECT in a patient with schizophrenia who had lost responsiveness to clozapine alone. We suggest further investigation to define the role of combined clozapine-ECT treatment in the management of treatment-resistant schizophrenia. PMID- 11941160 TI - ECT in a Patient with Harrington Rods. AB - A 40-year old man with severe psychosis and unresponsive to pharmacotherapy was treated with ECT despite the presence of Harrington Rods implanted after a suicide attempt. Complete relaxation was obtained with succinylcholine chloride 2.3 mg/kg and he received nine ECT without any musculoskeletal complications, nor did recovery from the treatments appear to be delayed. PMID- 11941162 TI - Double Stimulation to Achieve Adequate Treatment. PMID- 11941163 TI - Safe Administration of ECT in a Patient Taking Selegiline. PMID- 11941164 TI - Dental Consultation in ECT. PMID- 11941165 TI - Catatonia and DSM-IV. PMID- 11941166 TI - Catatonia in a University Inpatient Service (1985-1990). AB - Catatonia is defined only as a subtype of schizophrenia in the Diagnostic and Statistical Manual of Mental Disorders, 3rd edition (DSM-III) and DSM-III-revised (DSM-III-R), but catatonic symptoms are prominent in patients with psychoses, affective disorders, systemic illnesses, and neurotoxic states. We reviewed the case records of a 30-bed University Hospital adult inpatient psychiatric unit between 1985 and 1990, and identified 43 cases with admission or discharge diagnoses of schizophrenia, catatonic type (295.2). Of these, the records were adequate for detailed review in 19 cases. On discharge, seven were classified as schizophrenia, seven as affective disorder, and five as organic affective disorder. Eleven patients underwent electroconvulsive therapy (ECT), with excellent results in eight; 34 psychotropic medication trials were administered, with successful results in only two patients. Catatonia should be defined as a separate class in DSM-IV, thus encouraging the proper diagnosis and management of a treatable condition. PMID- 11941167 TI - The Response to an Amobarbital Interview as a Predictor of Therapeutic Outcome in Patients with Catatonic Mutism. AB - Intravenous amobarbital temporarily relieves catatonic mutism. The older psychiatric literature reported that response to amobarbital is associated with eventual good therapeutic outcome, whereas nonresponse may indicate poor outcome. This study investigated the short-term outcome of 20 patients with catatonic mutism after an amobarbital interview. A blind review of the charts was conducted at the patients' discharge, and patients were rated as either having good or poor outcome. The patients who did not respond to amobarbital improved during hospitalization as often as those patients who did respond to amobarbital. We conclude that a negative amobarbital interview is an unreliable prognosticator of poor response in these patients. PMID- 11941168 TI - ECT-Induced and Drug-Induced Hypomania. AB - In a retrospective chart study of 1,057 hospital admissions of endogenous depressives between 1920 and 1981, 139 patients (13%) had received electroconvulsive therapy (ECT), and 12% of them switched to hypomania. Within a subgroup of 524 psychotic unipolar depressives, 79 received ECT and eight (10%) switched to hypomania, whereas among those not treated with ECT only 16 of 445 patients (3.6%) switched to hypomania (p < 0.01). In psychotic bipolar patients the switch rates with and without ECT did not differ significantly (30% vs. 32%). Among untreated unipolar depressives hospitalized between 1920 and 1943, before the introduction of ECT or antidepressants, 3.9% switched to hypomania. Among unipolar patients admitted after 1957 and treated by antidepressants 4% switched to hypomania; among bipolar patients, 31% switched to hypomania. We find no evidence for hypomania being induced by standard antidepressants. Without classifying depressive patients into manic and nonmanic based on the previous history, studies of drug-induced hypomania cannot be conclusive. These observations strongly support the hypothesis of an ECT-induced switch from depression to hypomania. PMID- 11941169 TI - ECT During Pregnancy: Physiologic and Pharmacologic Considerations. AB - ECT is an accepted treatment for depression, mania, catatonia, and schizophrenia, but is usually used after other traditional drug treatments have failed. This also holds true for pregnant women who require psychiatric treatment. ECT is normally deferred until the fetus is at risk from the unstable psychiatric condition of the mother. This review examines the physiological effects of ECT, case reports of pregnant women receiving ECT, and possible complications of the medications administered before the procedure. All published case reports of ECT during all trimesters of pregnancy and the effects of ECT on the child are reviewed. The medications given before ECT (i.e., anticholinergics, muscle relaxants, and anesthetics) are examined for their teratogenic potential when administered multiple times during pregnancy. PMID- 11941170 TI - Dysarthria During ECT Given for Parkinson's Disease and Depression. AB - A patient with recurrent major depression, parkinsonism, and pseudobulbar symptoms underwent a course of electroconvulsive therapy. Although his depression and parkinsonian symptoms markedly improved, his dysarthria worsened, resulting in cessation of further treatments. Possible mechanisms are explored. PMID- 11941171 TI - Electroconvulsive Therapy in Parkinson's Disease: A Report on Five Cases. AB - Five patients with Parkinson's disease (PD) and depression underwent electroconvulsive therapy (ECT). One patient had a myocardial infarction several hours after the last planned session and died. Of the remaining four patients, all achieved marked improvement in mood and three had markedly improved motor function as well. Improvement in motor function lasted as long as 8 weeks. ECT is helpful in treating both depression and parkinsonism but maintenance ECT is required for continued motor improvement. PMID- 11941173 TI - Ethics of Electroconvulsive Therapy. PMID- 11941172 TI - Lateral Patient Position for Unilateral ECT. PMID- 11941174 TI - Weekly ECT in Geriatric Depression. AB - Fifteen elderly depressed psychiatric inpatients were randomly assigned to receive either standard three-times-weekly electroconvulsive therapy (ECT) or once-weekly ECT. Outcome measures included cognitive assessment and antidepressant response. Although both groups improved with treatment, the three times-weekly group improved substantially more quickly. There was no difference in cognitive effect between the two groups. We conclude that the traditional three-times-weekly schedule of ECT may optimally balance speed of antidepressant response and cognitive impairment. PMID- 11941175 TI - Tissue Injury-Inducing Potential of Unmodified ECT: Serial Measurement of Acute Phase Reactants. AB - Eight subjects with acute functional psychoses receiving unmodified electroconvulsive therapy (ECT) at Ibadan were investigated for occult or subclinical internal tissue damage by serial measurements of eight acute phase reactants. Samples of venous blood were collected from each patient at pretreatment, two within treatment, and one at posttreatment. The acute phase proteins assayed were C-reactive protein (CRP), alpha-2-macroglobulin, ceruloplasmin, factor B, C-4 protein, C-3 protein, transferin, and alpha-1 antitrypsin. Except for CRP, the values of the proteins did not change during treatment. CRP values decreased posttreatment and were not detectable in the last sample in five subjects in whom values had been present pretreatment. Our data do not support fears of occult internal tissue damage during unmodified ECT. The consistent decrease in CRP levels posttreatment when patients no longer exhibited psychotic symptoms could not be explained by type of psychosis, intramuscular injections, or changes in drugs and diet; its significance is not known. PMID- 11941176 TI - Electroencephalographic Seizure Duration in Electroconvulsive Therapy: A Clinical Study. AB - Electroencephalographic estimates of electroconvulsive therapy (ECT)-induced seizure duration were obtained in 141 patients with major depression (n = 41) or schizophrenia (n = 100). A slight reduction in seizure duration across a course of ECT failed to reach statistical significance and was unrelated to clinical improvement. PMID- 11941177 TI - Propofol Anesthesia in ECT. AB - Multiple independent investigators have reported that propofol anesthesia during electroconvulsive therapy (ECT) is associated with substantially shorter ECT seizure duration than barbiturate anesthesia, which evidences seizure obstruction by propofol. Seizure length after propofol narcosis is often below the 20-30 s range accepted as minimally adequate. Seizure-induced elevations of prolactin, adrenocorticotropic hormone, and cortisol are similarly lower after propofol. Together with a recent report that average course length with propofol exceeds 16 sessions, this report indicates that propofol is undesirable for routine ECT anesthesia, and also illustrates how aspects of ECT anesthesia that affect the psychiatric or behavioral outcome require the collaboration of a psychiatrist. PMID- 11941178 TI - Treatment Choice in Neuroleptic Malignant Syndrome. AB - A literature review of patients with symptomatic neuroleptic malignant syndrome (NMS) treated with electroconvulsive therapy (ECT) yielded 26 cases, to which we add five cases. ECT was associated with a positive outcome in 26 of 31 cases with one unclear outcome and a poor outcome in four cases, including two deaths. ECT appeared to be effective in eight of nine patients previously treated with dantrolene and/or bromocriptine; no difference in time to apparent response was seen between those treated with medication first and those undergoing ECT first. The mean time to clinical response after the first ECT was 1.46 +/- 2.38 days, with 19 of 20 having a clinical response by 72 h. The possible relationship of ECT to the two deaths is discussed. Given this experience, the suggested treatment sequence is medication (dantrolene or bromocriptine) for 48 h; if no clinical response is seen, ECT should be initiated. ECT may be used earlier in response to specific clinical situations. PMID- 11941179 TI - Heterozygous Atypical Cholinesterase: Management During ECT. AB - A 70-year old woman developed prolonged apnea during an initial administration of electroconvulsive therapy (ECT) with succinylcholine. Because of the duration of apnea, she was suspected of having a heterozygous type of atypical cholinesterase. Subsequent therapies were administered with a reduced dose of succinylcholine. Prolonged apnea did not recur. PMID- 11941180 TI - Post-ECT Myoclonic Jerks in a Depressed Patient with Bulimia. AB - We describe an individual with psychotic depression and bulimia who demonstrated spontaneous post-electroconvulsive therapy (ECT) myoclonic jerks after her fifth unilateral treatment. After switching to bilateral ECT, the patient developed generalized tonic-clonic seizures, beginning about 2 min after each treatment and lasting for about 4 min. These were confirmed by electroencephalographic (EEG) recording, and could be terminated and also effectively prevented by intravenous midazolam. Differential diagnosis, risk factors, and management are discussed. PMID- 11941181 TI - Electroconvulsive Therapy in a Depressed Heart Transplant Patient. AB - Medication-resistant major depression was diagnosed in a 67-year-old man 1 1/2 years after heart transplantation. Electroconvulsive therapy was administered, resulting in remarkable improvement in depressive symptoms without complications related to the cardiac condition. PMID- 11941182 TI - Electrode Placement and Antidepressant Efficacy. PMID- 11941184 TI - Post-Electroconvulsive Therapy Shaking. PMID- 11941183 TI - Electroconvulsive Therapy-Induced Hypomania is Uncommon. PMID- 11941185 TI - Beyond Seizure Duration as a Measure of Treatment Quality. PMID- 11941186 TI - Maintenance Electroconvulsive Therapy and Seizure Duration. AB - A retrospective study was conducted of 14 affectively ill patients who were treated with long-term unilateral maintenance electroconvulsive therapy (MECT). The patients had received an inpatient course of ECT before being referred for MECT. The average age was 57 +/- 16 years (range 30-91). The average interval between inpatient ECT was 2.4 +/- 0.9 days, in contrast to the interval between MECT, which was 12.1 +/- 11.3 days. The average time to start MECT after inpatient ECT was 16.2 +/- 16.2 days. The average duration of MECT was 81 +/- 104 days (maximum 571). Patients' affective symptoms continued to improve during the course of MECT based on Carroll Depression Ratings. Adjustment of the electrical dose and caffeine augmentation were used to keep the seizure durations >30 s by electroencephalograph (EEG) monitoring. Over time, most treatments were administered using the maximal charge provided by the Mecta SR-1. Despite considerable time intervals between MECT treatments, seizure durations did not increase. Additionally, high stimulus charge and frequently administered caffeine were used to maintain seizure length. The apparent anticonvulsant effect of ECT was not lost over the time span of MECT. This has clinical implications if the anticonvulsant effects of ECT contribute to determining the clinical response. PMID- 11941187 TI - Blood Pressure, Memory, and Electroconvulsive Therapy. AB - Blood pressure changes recorded during electroconvulsive therapy (ECT) in 23 psychiatric in-patients with major depressive disorders correlated with and predicted the degree of anterograde memory changes measured 48-72 h after ECT. The Randt memory test was the principal measure of memory change. A subgroup of older patients with cardiovascular illness received trimethaphan, a ganglionic blocker that impedes a hypertensive surge during the treatment. They did not differ in memory function from a younger subgroup that did not receive trimethaphan. Control of the hypertensive response in the older age group counterbalanced the additional memory dysfunction that was anticipated as a result of advanced age and cardiovascular pathology. PMID- 11941188 TI - Effect of Stimulus Energy on Electroconvulsive Therapy-Induced Prolactin Release. AB - This study was undertaken to examine the effect of stimulus energy on electroconvulsive therapy (ECT)-induced prolactin (PRL) release. Patients undergoing a course of right unilateral or bilateral ECT were studied during two consecutive treatments. The order of high- and low-energy ECT was counterbalanced between the two treatments. A multivariate repeated-measures analysis of variance with seizure duration within each condition (high/low energy ECT) used as a covariate showed that seizure duration was not significantly different between conditions, but was significantly related to PRL release. However, PRL increase in response to high-energy ECT was significantly higher compared with low-energy ECT. These results indicate that in addition to the previously demonstrated relationship between electrode placement and ECT-induced PRL response, PRL release also varies as a function of stimulus energy. The implications of the hypothesis that this neuroendocrine response could serve as an indication that an effective stimulus has been delivered are discussed. PMID- 11941189 TI - The Randt Memory Test in Electroconvulsive Therapy: Relation to Illness and Treatment Parameters. AB - We used the Randt Memory Test (RMT) to examine the effects of electroconvulsive therapy (ECT) on memory in 23 depressed patients. Immediate recall was unaffected by ECT, but delayed recall was reduced 48-72 h following reorientation after ECT. These changes were not correlated with the total seizure time or with the dose of electrical energy. Most patients rated their memory as improved after ECT on the Squire Subjective Memory Questionnaire. Improvement in mood, as reflected in behavior scales, was correlated with the subjective perception of memory performance, but not with the objective performance on the RMT. Improvement in depressive mood is independent of changes in performance on tests of memory. The RMT is a convenient and useful instrument to monitor the memory effects of ECT. PMID- 11941190 TI - Age-Dependent Effects of Electroconvulsive Therapy on Memory. AB - We examined the relation between age and recovery of memory functions after electroconvulsive therapy (ECT). In a group of patients 20-65 years of age, older depressed patients treated with ECT experienced more severe and longer lasting memory deficits than did younger patients. Testing conducted 24-72 h after a course of ECT showed more severe deficits in older patients for verbal and visuospatial anterograde memory, and for retrograde memory. The difference between younger and older subjects was marginal at 1 month follow-up, seen only in differences in verbal anterograde memory. At 6 months follow-up, no difference in memory test scores between older and younger patients was observed. Older patients are more vulnerable to cognitive effects of ECT, and these effects last longer. PMID- 11941191 TI - Malignant Hyperthermia and Electroconvulsive Therapy. AB - A 30-year-old man with a personal and family history of malignant hyperthermia and a 7-year history of psychiatric illness unresponsive to various psychotropic medications benefitted from electroconvulsive therapy given in combination with clozapine. Volatile inhalation anesthetics and a depolarizing muscle relaxant (succinylcholine) were assiduously avoided. Dantrolene was administered intravenously before the first treatment but was not used for the remainder of the treatments. Anesthesia was induced with methohexital and atracurium. The treatment course was uneventful. PMID- 11941192 TI - Electroconvulsive Therapy in Mania: Successful Outcome Despite Short Duration of Convulsions. AB - A duration of 25-30 s of convulsions in electroconvulsive therapy (ECT) is thought to be the minimum necessary for an adequate treatment. We describe three manic attacks in a 48-year-old patient resistant to psychotropic drugs who responded to successive courses of ECT, each with unusually short durations of seizures. PMID- 11941193 TI - Electroconvulsive Therapy and Lithium: Safe and Effective Treatment. AB - It is often recommended that lithium not be prescribed during electroconvulsive therapy (ECT). The combined use of these treatments has been considered a risk factor for inducing encephalopathy; however, this precaution is controversial. We report the case of a woman who received over 70 outpatient maintenance ECT concurrently with lithium for years without signs of confusion. Similar observations are reported by others, too. The relative merits of all therapeutic interventions should be individualized to each patient. PMID- 11941195 TI - Prolonged Seizures. PMID- 11941194 TI - Electroconvulsive Therapy in Creutzfeldt-Jakob Disease. AB - Creutzfeldt-Jakob disease is a degenerative disorder of the central nervous system that frequently presents with behavioral symptoms including depression. We report here a patient with Creutzfeldt-Jakob disease in whom electroconvulsive therapy (ECT) was highly effective in relieving depressive symptoms. Additionally, ECT had little or no effect on cognition in this patient, even transiently improving her cognitive function. Given its rapid onset of action, ECT may be considered safe and effective in depressive disorders associated with Creutzfeldt-Jakob disease. PMID- 11941197 TI - Caffeine Pretreatment Enhances Clinical Efficacy and Reduces Cognitive Effects of Electroconvulsive Therapy. AB - In an open clinical trial, depressed patients received age-dosed, brief-pulse electroconvulsive therapy (ECT) either with or without 500 mg i.v. caffeine sodium benzoate before each treatment. Caffeine-pretreated patients required fewer ECT treatments, and after three to four treatments, their Hamilton Depression Scale (HDS) scores were significantly lower. At the end of the ECT course, both groups reached the same reduction in HDS scores. Of five memory tests, one showed better performance at the end of the ECT course for the caffeine-pretreated compared with the non-caffeine-pretreated patients. The results argue that caffeine-modified ECT differs from unmodified ECT in speed of response and the effects on cognitive tests. PMID- 11941196 TI - Electroconvulsive Therapy Stimulus Dosing: A Survey of Contemporary Practices. AB - There is debate in the psychiatric community regarding how to optimize electroconvulsive therapy (ECT). American Psychiatric Association recommendations suggest a moderately suprathreshhold stimulus intensity, yet little information is available regarding how practitioners select initial ECT stimulus doses. We report the results of a nationwide study of common ECT practices. Respondents provided their most commonly used electrode placement, method of determining stimulus intensity, and type of ECT device. Our results showed a high degree of variability in ECT methods, and those practitioners "specializing" in ECT were no more likely than nonspecialists to titrate stimulus intensity at the first treatment. Indeed, a fixed high-dose stimulus, stimulus titration, and stimulus intensity calculated by formula were each widely used by both ECT specialists and nonspecialists. PMID- 11941198 TI - Electroconvulsive Therapy-Related Psychiatric Knowledge Among British Anesthetists. AB - Opinion of electroconvulsive therapy (ECT) and ECT-related psychiatric knowledge among anesthetists working in the North Western Health Region of England were investigated; 261 anesthetists returned usable questionnaires. The majority were in favor of ECT for patients as recommended by psychiatrists, and possessed a reasonable knowledge base about related psychiatric issues. A minority were opposed to the use of ECT, lacked rudimentary psychiatric knowledge and held negative misconceptions. Although this does not compromise anesthetic ability, it may have adverse consequences. In particular anesthetists may not appreciate the importance of their role in ECT and may be reluctant to become involved in ECT clinics and research. Attitude and knowledge did not differ significantly when respondents with extensive and recent ECT experience were compared with the remainder. Two thirds of respondents favored anesthetic training including more information about psychiatric aspects of ECT. We recommend improved liasion between anesthetists and psychiatrists during training and in clinical practice. PMID- 11941199 TI - Cardiovascular Response and Anesthetic Recovery in Electroconvulsive Therapy with Propofol or Thiopental. AB - Propofol provokes a slight hypotensive effect that could mitigate the cardiovascular response to electroconvulsive therapy (ECT). In this study we compared the effects of propofol and thiopental for ECT anesthesia in seven women (22-67 years of age). Anesthesia was induced with either thiopental or propofol, and with atropine and suxamethonium for each treatment. The first anesthesia was assigned to thiopental or propofol at random; the next anesthesia was induced with the other drug, and alternated thereafter. Systolic blood pressure, diastolic blood pressure (DBP), and heart rate (HR) were recorded before anesthesia, after anesthetic induction, and 1 and 5 min after ECT. ECT-induced increases in DBP and HR were less marked with propofol than with thiopental. Seizure durations were decreased with propofol compared with thiopental. PMID- 11941201 TI - Aortic Aneurysm and Electroconvulsive Therapy. AB - We report two patients with confirmed aortic aneurysm who successfully received ECT. A review of the literature on ECT finds 13 prior case descriptions. Based on the reported cases and literature review, we conclude that ECT in patients with aortic aneurysm is a safe and effective procedure. Although patients may benefit from additional medication for blood pressure control, the evidence suggests that they may be safely treated without invasive monitoring. PMID- 11941200 TI - A Multivariate Analysis of the Experience of 423 Depressed Inpatients Treated with Electroconvulsive Therapy. AB - Stepwise multiple logistic regression was used in an attempt to identify variables associated with "recovery" in a group of 423 depressed inpatients who received electroconvulsive therapy (ECT). Patients who had recovered were older at index admission than the nonrecovered, and had received fewer ECT sessions. They were less likely to have a secondary depression, to have Winokur's depression spectrum disease, to receive lithium, or to have a chronic course. The significance of the findings is discussed. PMID- 11941202 TI - Nonconvulsive Status Epilepticus after the Ninth Electroconvulsive Therapy. PMID- 11941203 TI - Seizure Duration and Seizure Threshold. PMID- 11941205 TI - Ethics and Consent. PMID- 11941204 TI - Authors' Reply. PMID- 11941206 TI - Author's Reply. PMID- 11941207 TI - Electroconvulsive Therapy in Children and Adolescents. PMID- 11941208 TI - Electroconvulsive Therapy in Adolescents. AB - Because of the limited number of case reports on the use of electroconvulsive therapy (ECT) in adolescent psychiatric patients, we retrospectively reviewed the medical records of 20 young patients who underwent ECT. They constituted all the patients /=20%) in the Brief Psychiatric Rating Scale total scores by the end of the course of ECT. Improvement was greater in positive than in negative symptoms. A sixth patient improved 3-6 weeks after the completion of the acute course of ECT. Five patients who responded to ECT received maintenance ambulatory ECT; two patients remained improved for at least 6-9 months, whereas the other three relapsed within 4 months. Further study of the indications for and the effectiveness of ECT plus neuroleptic drugs in treatment-resistant schizophrenia is indicated. PMID- 11941210 TI - Combined Clozapine and Electroconvulsive Therapy. AB - We reviewed consecutive patients (n = 12) at McLean Hospital from 1990 through 1991 treated with the combination of the atypical antipsychotic agent clozapine and electroconvulsive therapy (ECT). There were no adverse effects. Three patients had a marked clinical improvement, one a moderate response, four a minimal response, two minimal to no response, and two no response. Using daily doses of up to 550 mg clozapine, this combination appears to be safe, and may be useful in some patients with treatment-refractory psychosis. PMID- 11941211 TI - Age Effect on Electroconvulsive Therapy Seizure Resistance. AB - To specifically examine the expectation that seizure induction is more difficult in older patients, we studied a consecutive series of 21 men receiving a course of brief-pulse, square-wave, bilateral electroconvulsive therapy (ECT). Each subject received a 144-mC stimulus over 1 s (800-mA, 1.5-ms pulses at 60 Hz) at the second, third, or fourth ECT session, randomly selected. Nine subjects showed no convulsion. Subjects who showed seizure (age 37.7 +/- 13.2 years) were younger (t = 4.178, df = 19, p = 0.00025, one-tailed) than those who did not (age 57.6 +/ 6.1 years). These observations indicate that seizure resistance to bilateral brief-pulse ECT increases strongly with age in men. PMID- 11941212 TI - Sustained Downregulation of Cortical Adrenergic Receptor Density with Maintenance Electroconvulsive Stimulation. AB - Central beta-adrenergic receptor downregulation is a robust and reversible neurochemical finding in animals after a course of electroconvulsive stimulation (ECS) or antidepressant drug administration. In addition, beta-receptors are essential components of monoamine-based models of affective disorder pathogenesis and treatment. We tested whether the downregulation in rat cortical beta-receptor density observed after an initial series of ECS is maintained over 2 months of maintenance ECS. Our findings confirm prior reports, in the absence of maintenance ECS, that receptor density is decreased after a course of 10 daily ECS, with recovery to baseline 16 days after the last ECS. In contrast, when maintenance ECS was given for 2 months after an initial series of 10 ECS, a schedule of maintenance ECS at two per week was sufficient to sustain the receptor downregulation. The results are discussed in terms of the neurochemical study of ECS in animals and implications for maintenance ECS. PMID- 11941213 TI - Neuroleptic Malignant Syndrome: Neuroleptic Rechallenge After Electroconvulsive Therapy. AB - The necessity of neuroleptic rechallenge in patients who have experienced neuroleptic malignant syndrome (NMS) has been discussed in recent articles. Electroconvulsive therapy (ECT) has been proposed as an alternative treatment to neuroleptics. In cases where ECT fails, neuroleptic rechallenge remains the only alternative. A case of NMS is reported with successful reintroduction of neuroleptics after an ECT course. Four prior rechallenges with neuroleptics before ECT had failed, and NMS recurred. ECT may change the reaction to neuroleptics. PMID- 11941214 TI - Electroconvulsive Therapy in Patients with Cerebral Palsy. AB - Recent reviews attest to the safety and efficacy of electroconvulsive therapy (ECT) in psychiatrically ill patients with concomitant neurologic disease. Four patients with mood disorders and cerebral palsy are described who achieved a therapeutic response with ECT without neurologic deterioration. ECT is safe in the treatment of mood disorders in patients with cerebral palsy. PMID- 11941215 TI - Reversible Dementia and Affective Disorder: The Rip Van Winkle Syndrome. AB - The diagnostic separation of the reversible dementia of an affective disorder from the dementia secondary to structural brain pathology remains a clinical challenge. A 58-year-old woman had been diagnosed as having Alzheimer's dementia for 9 years before antidepressant treatment with electroconvulsive therapy (ECT) resolved the dementia syndrome. The patient has functioned well for 8 years on maintenance treatment with lithium, with ECT given every 7-8 weeks. By the summer of 1993, she had undergone 132 ECT. Until specific and reliable pre-morbid tests for the diagnosis of irreversible dementias of the Alzheimer's and multiinfarct types are developed, antidepressant treatment trials are encouraged in elderly patients with a dementia syndrome. Extensive maintenance ECT schedules are safe. PMID- 11941216 TI - Electroconvulsive Therapy for Depression After Temporal Lobectomy for Epilepsy. AB - Electroconvulsive therapy (ECT) was administered to a 25-year-old woman with major depression with psychotic features who had undergone a left temporal lobectomy for medically intractable partial epilepsy 1 year earlier. Her depressive illness responded to ECT without ill effects to her seizure disorder. PMID- 11941217 TI - Low-Frequency Ictal EEG Activity and ECT Therapeutic Impact. PMID- 11941218 TI - Author's Reply. PMID- 11941219 TI - ECT and Drugs: Concurrent Administration. PMID- 11941220 TI - Combining ECT and Antipsychotic Agents: Benefits and Risks. AB - A comprehensive review describes the benefits and risks of the combination of electroconvulsive therapy (ECT) and antipsychotic agents for patients with psychotic disorders. For some patients, combined treatment may lead to greater rates of response or more rapid improvement than with either therapy alone. Except for reserpine, combining neuroleptics with ECT is safe, but some precautions are suggested. The mechanism of interaction, effect on seizure threshold, the use of ECT for neuroleptic associated motor effects, and additional anesthesia considerations are discussed. PMID- 11941221 TI - Combined ECT and Antidepressant Drug Therapy. AB - The current literature on the interaction between antidepressant drugs and electroconvulsive therapy (ECT) is reviewed. Tricyclic antidepressants, atypical antidepressants, and monoamine oxidase inhibitors are discussed. The efficacy of combinations and possible adverse effects (mainly cardiovascular and seizure threshold effects) are considered. Many previous studies on the efficacy of combinations used inadequate methods, and the safety data consist largely of anecdotes and small case series. Additional studies are needed to assess the safety and efficacy of the combined use of antidepressants and ECT. PMID- 11941222 TI - Benzodiazepines and Electroconvulsive Therapy. AB - Benzodiazepines (BZD) possess anticonvulsant properties that may potentially elevate seizure threshold, inhibit seizure propagation, and alter some of the neurobehavioral effects of electroconvulsive shock (ECS) in animal models. Nevertheless, considerable controversy exists regarding the clinical impact of oral BZD use during electroconvulsive therapy (ECT). The existing literature is contradictory, and all studies attempting to address this topic suffer from important design flaws. Most studies are retrospective and some address only seizure duration. Also, studies examining treatment outcome are difficult to compare because of differing types and dosages of BZD, varied electrode placement and stimulus energy, and lack of information about the relationship of the stimulus energy to the patients' seizure thresholds. While firm conclusions must await further studies, limited data suggest that BZD have the potential to shorten seizure duration and decrease treatment efficacy, particularly with unilateral ECT. PMID- 11941223 TI - Combined ECT and Lithium Therapy. AB - Current guidelines for clinical practice advise against the concurrent use of lithium during a course of electroconvulsive therapy (ECT) owing to an increased risk of morbidity involving the central nervous system. A reappraisal of the evidence indicates that the hazards of this combination treatment may have been overestimated. Although the potential therapeutic advantages of the combination treatment over ECT or lithium alone remain to be clarified, in special clinical situations, and with proper precautions, ECT-lithium combination may be used safely and effectively as an alternative treatment strategy where conventional therapeutic approaches have failed. PMID- 11941224 TI - Combined Use of Thyroid Hormone and ECT. AB - Neurocognitive deficits, including acute confusion and memory impairment, remain important effects of electroconvulsive therapy (ECT). Laboratory and clinical research demonstrates interactions among neurocognitive functioning, the hypothalmic-pituitary-thyroid axis, depressive mood, and ECT. Preclinical studies have demonstrated that exogenous triiodothyronine (T(3)) administered to animals receiving electroconvulsive shock (ECS) protects against ECS-related amnesia and accelerates the "antidepressant effects" of ECS, possibly due to alterations in catecholamine and/or indoleamine neurotransmission. Clinical data suggest that combined treatment with T(3) and ECT results in increased clinical efficacy of ECT and diminished neurocognitive side effects. PMID- 11941225 TI - Anticholinergics and ECT. AB - Anticholinergic medication is commonly used prior to anesthetizing patients for electroconvulsive therapy (ECT). Its routine use is the subject of ongoing controversy. This article reviews how anticholinergics came to be used with ECT and the work supporting or refuting their routine use. The route of administration and their use with beta blockers are reviewed. Atropine, glycopyrrolate, and scopolamine are compared. While acknowledging the pau-city of carefully controlled trials with large numbers of patients, the routine use of intravenous atropine prior to administering ECT is recommended unless specifically contraindicated. PMID- 11941226 TI - Anesthesia for ECT. AB - This article reviews the use of sedatives and narcosis agents, muscle relaxants, and ventilation that are specific to ECT anesthesia rather than surgical anesthesia. Particular focus is given to interference with a seizure by pretreatment sedation and anesthetic narcosis; dosage of succinylcholine and alternatives to succinylcholine; emergence agitation; and the effects of ventilation on seizure quality. PMID- 11941227 TI - Antihypertensive Medications and ECT. AB - Prominent changes in hemodynamics occur during electroconvulsive therapy (ECT) despite general anesthesia and muscle relaxation and regardless of choice of stimulating electrode placement. The most common pattern is hypertension and tachycardia during the seizure, followed by relative bradycardia and a gradual return of blood pressure to normal within minutes of the end of the seizure. The mortal and morbid risk of these changes is generally small; nevertheless, it is common practice to administer antihypertensive medications to blunt this response. Several agents have proven able to reduce ECT-related hypertension and tachycardia; however, it is unknown whether these agents reduce mortality or cardiovascular morbidity or alter efficacy or cognitive effects of ECT. A few case reports suggest intravenous antihypertensives may be positively dangerous on occasion. Based upon these findings, antihypertensives need not be given routinely during ECT, and when given, conservative doses should be used. The short-acting beta blockers may be particularly suited to this task. To prevent needless use of these agents and the possibility of an adverse event, the patient's intravascular volume, blood pressure and pulse should be optimized prior to the first ECT. PMID- 11941228 TI - Post-ECT Delirium. AB - A confusional state accompanied by restlessness is a frequent occurrence at the end of an induced seizure in electroconvulsive therapy (ECT). Delirium, which usually lasts <1 h, occurs in up to 12% of patients. It is most frequent during the initial treatments of a series. The occurrence of delirium in one treatment is not a harbinger of its reappearance in later treatments. Usually gentle reassurance is sufficient to help the patient, but some instances warrant the administration of diazepam, midazolam, a barbiturate, or droperidol. PMID- 11941229 TI - Augmentation Strategies in Electroconvulsive Therapy: A Synthesis. AB - Over the past four decades, a variety of pharmacological augmentation strategies have been used in electroconvulsive therapy (ECT). Their goal has been to enhance therapeutic effects and seizure manifestations, or to minimize cognitive effects. Although they are not intended to affect outcome or other clinical parameters, medications taken during ECT may also impact on safety and efficacy. There is a large literature in epileptology on how pharmacologic agents affect seizure physiology. We review the clinical literature on augmentation strategies in ECT, with a synopsis of relevant preclinical studies, and seek to relate the pharmacological effects on seizure parameters to critical clinical variables in ECT. PMID- 11941230 TI - [Vitamin E: biochemical, clinical and epidemiological aspects] PMID- 11941231 TI - [Clinical evaluation of the ocular safety of Amukine 0.06% solution for local application versus povidone iodine (Betadine) 5% solution for ocular irrigation) in preoperative antisepsis]. AB - INTRODUCTION: The aim of this monocentric, randomized, comparative, open study was to evaluate the safety of Amukine 0.06% solution (an isotonic hypochloride sodium solution) versus a 5% povidone iodine solution (Betadine 5% solution for ocular irrigation) in antisepsis before cataract surgery. MATERIAL AND METHODS: One hundred and thirteen patients, aged between 49 and 90 years, were included and split, after randomization, into two groups: one testing Amukine 0.06%, the other testing a 5% povidone iodine solution. For each group, after local anesthesia, the antiseptic procedure consisted of an antisepsis of periocular teguments followed by an eye antisepsis through the instillation of two drops of the tested product in the conjunctival fornices. The safety evaluation was performed by grading superficial punctate keratitis 24 hours after surgery (slit lamp examination after fluorescein instillation). Conjunctival hyperemia scores by examination of the bulbar conjunctiva before the first instillation, immediately before surgery, and 24 hours after surgery were also compared. RESULTS: One hundred and seven reports were analyzed. Concerning the presence of corneal superficial punctate keratitis 24 hours after surgery, mean scores were not different (0.27 for the Amukine 0.06% group and 0.38 for the povidone iodine group; p=0.27 Mann Whitney test). The observations did not show a different progression of conjunctival hyperemia depending on the treatment group (p=0.65 (immediately after application) and p=0.52 (after 24 hours); Mann Whitney test). CONCLUSION: In this study, the ocular safety of Amukine 0.06% solution was not different from a 5% povidone iodine solution. Therefore, Amukine 0.06% is a new interesting approach to surgical antisepsis in ophthalmology and an alternative in case of iodine allergy. PMID- 11941232 TI - [Posttraumatic glaucoma]. AB - INTRODUCTION: Ocular trauma is an important cause of blindness among young people. Injuries of the iridocorneal angle cause ocular hypertonia and postcontusive glaucoma. The purpose of this work is to study clinical and therapeutic particularities of postcontusive ocular hypertonia. METHODS: This is a retrospective study on 13 patients (12 men and 1 woman) with ocular hypertonia as a result of contusive trauma of the iridocorneal angle, with no hyphema or lens dislocation. Patients were 16 to 38 years old. RESULTS: Ocular hypertonia appeared between 1 week and 6 months after the trauma. The lesions involved were cyclodialysis (2 cases), angle recession (11 cases) and iridodialysis (2 cases). Ocular hypertonia was complicated in 6 cases by contusive glaucoma (as shown by optic disk examination and visual field exploration). The balancing of intraocular pressure was obtained by local treatment in only 9 cases and by trabeculectomy in 5 cases. DISCUSSION: The authors discuss the physiopathology, therapeutic particularities as well as the prognostics of contusive ocular hypertonia by iridocorneal angle damage. CONCLUSION: Screening for ocular hypertonia must be regular and systematic after ocular trauma involving lesion of the iridocorneal angle. PMID- 11941233 TI - [Evaluation of single-injection caruncular sub-Tenon's anesthesia]. AB - INTRODUCTION: Because of its complications, particularly globe perforation, retrobulbar injection has been progressively replaced by peribulbar anesthesia. However, with peribulbar anesthesia, the excessive rate of imperfect blockade requires supplemental injection. We have been performing local anesthesia using caruncular sub-Tenon single injection for many years. This technique is evaluated. PATIENTS AND METHODS: A prospective study has been carried out on 183 eyes. Regional anesthesia was given by a caruncular sub-Tenon single injection. For each case, we studied akinesia, analgesia, pain before, during, and after surgery, the number of supplemental injections, eyeball orientation, and surgical conditions. RESULTS: A total motor blockade was obtained in 157 cases (85.8%) and total lid akinesia in 176 patients (96.2%). Eight patients (4.4%) needed a second injection. During surgery, 10 patients complained of pain (5.5%). We found 27 eyes (14.7%) were divergent and 12 (6.5%) were convergent. Surgical conditions were good or very good in 90% of cases. No complications due to the injection (perforation or lesion of the eyeball or the optic nerve) were noted. CONCLUSION: Single-injection caruncular sub-Tenon anesthesia is an alternative technique to classical peribulbar anesthesia. This technique is efficient, simple, easy to learn, reproducible, and seems to have a low rate of complications. PMID- 11941234 TI - [First line endonasal dacryocystorhinostomy Technique and results. Comparison between diode laser and electrocautery instrument. Study based on 422 procedures]. AB - PURPOSE OF THE STUDY: To assess the results of the first procedures of endoscopic endonasal dacryocystorhinostomy (ENL DCR). To study the efficiency of diode laser in this operation versus electrocautery instruments (ECI). DESIGN: Prospective study; comparative study. METHODS: Intraoperative comparison between ECI and diode laser is based on 422 ENL DCR achieved between June 1997 until June 2000. Three hundreds and sixty three procedures were diode laser assisted and 59 procedures were done with the use of an ECI. The diode laser was only used to vaporize nasal mucosa laying on the osteotomy site, to realize partial turbinectomy and to vaporize polyps and synechiae. Intraoperative comparison was based on the followings: frequency and extent of bleeding, quality of visibility, perception of pain during the use of the instrument and duration of the operation. Postoperative comparison is based on 318 procedures. Comparison was based on: frequency of granuloma formation in the nasal mucosa at the site of the osteotomy, frequency of acquired nasal synechiae, frequency of important crusting reaction of the nasal mucosa; success rates after a minimum follow up of 6 months. RESULTS: Intraoperative haemorrhages were fewer and smaller in the diode laser assisted procedures than in ECI assisted procedures; visibility was better with the diode laser; the use of the diode laser was painless versus 24% of patients complaining of pain during the use of electro-cautery. The duration of the procedure was shorter with the diode laser (29'30 vs 37'). In the postoperative follow-up frequency of granuloma formation was equal with the two instruments, synechiae were fewer with the diode laser (11% vs 22%) just like crusting reaction of the nasal mucosa (7% vs 36%). Success rates were similar (diode laser: 91.94% vs EC: 86.66%). CONCLUSION: Because of the effeciency of the vaporization and coagulation, the diode laser, in comparison with the ECI, allows to shorten mean operative time, to improve the visibility and to decrase bleeding. Contrary to EC it's use is painless. Fewer important crusting reactions and fewer synechiae after diode laser assisted procedures tend to prove than postoperative inflammation and surgical trauma are smaller than with ECI. There is no statiscally signifant difference between the success rates of ENL laser diode assisted procedures and ENL ECI assisted procedures. PMID- 11941235 TI - [Ocular involvement during primary central nervous system lymphoma]. AB - PURPOSE: The aim of our study was to assess and to characterize the ocular involvement in patients with primary central nervous system lymphoma. POPULATION AND METHODS: We conducted a prospective study between August 1995 and December 1998 on a cohort of consecutive patients affected by primary central nervous system lymphoma and who underwent a systematic ophthalmological examination before treatment. RESULTS: The study population comprised 24 patients (mean age, 57 years; 16 women and 8 men). Among these patients, 6 had ocular involvement (2 patients with ocular signs revealing the cerebral process, 2 evidenced at the time of the initial ocular examination, and 1 as the disease evolved). Ocular involvement was vitritis in all cases, anterior uveitis in 1 patient, obliterant vasculitis in 1 patient and yellowish subretinal lesions in 1 patient. In 1 case, a vitrectomy led to adequate diagnosis of the disease. CONCLUSION: Our study shows a 25% incidence of ocular involvement in patients with primary central nervous system lymphoma, with posterior uveitis as the most common manifestation. Because of the relatively high incidence of ocular involvement, a systematic ophthalmological examination of this subset of patients should be mandatory. PMID- 11941236 TI - [Vitreous luxated PC-IOLs: complications]. AB - PURPOSE: To determine pre-, intra- and postoperative complications of posterior chamber intraocular lenses (PC-IOL) posteriorly luxated in the vitreous. METHODS: We retrospectively reviewed reports of all patients with luxated PC-IOLs at our institution (1989-1999) with a minimum follow-up of 6 months. We identified 41 eyes; 4 were excluded because of short follow-up. Thirty-seven eyes were finally considered. Twenty-one eyes had "in situ" repositioning with vitrectomy, in 9 of them perfluorocarbon liquids (Perfluoro-DK-line) (PFCL) were used to refloat the luxated lenses. Sixteen eyes had IOL exchange, in 7 of them PFCL was used to refloat the IOL. RESULTS: The average age of the patients was 67.5 years. Preoperatively, 15 eyes (40%) had ocular hypertension, 9 eyes (24%) showed epithelial corneal edema (CE), 6 eyes (16%) had light vitreous hemorrhage (VH) and 4 eyes (11%) retinal detachment (RD). Intraoperatively, 9 eyes (24%) had VH related to sulcus fixation. Postoperatively, visual acuity improved in 32 (86%) eyes. In 3 cases (8%) we found a postsurgical RD that could be iatrogenic; one eye developed glaucoma and 1 developed, epiretinal membrane. Two patients were PFCL drops. CONCLUSION: Vitrectomy normalizes IOP and CE, PFCL simplifies the surgical technique for RD, and sulcus fixated IOL allows visual restoration. The combined technique generally offers good results. PMID- 11941237 TI - [Comparison between Greenbaum's parabulbar anesthesia and Ripart's subtenon anesthesia in the anterior segment surgery]. AB - PURPOSE: To compare two methods of sub-tenon anesthesia in 80 surgical procedures (phakoemulsification, glaucoma and combined surgery) in a prospective, single surgeon study. MATERIALS AND METHODS: Forty patients requiring anterior segment surgery in each eye were randomised to receive subtenon anaesthesia by either Greenbaum's method (using a flexible plastic cannula) or Ripart's method (using a 23G hypodermic needle). Randomization dictated the mode of anaesthesia for the first eye, the other technique being used for the second eye. Anaesthesia consisted of 1.5ml lidocaine 2% and 1.5ml bupivacaine 0.5% in all cases, and was performed by the same surgeon (EDLM) immediately before surgery. Type of surgical procedure, duration, complications, presence of sub-conjunctival haemorrhage, were assessed by the surgeon, who also graded chemosis (0-3), nuclear hardness (1 4), and ocular akinesia (0-2) for each patient. Pain was scored subjectively by each patient (0-10) during the injection, peroperatively and postoperatively. RESULTS: Chemosis was significatively higher with Greenbaum's method than Ripart's method (p<0.01) and was sometimes undesirable for the surgeon. There was no difference in the pain score during the injection, preoperatively or postoperatively. Pain was usually very light and did not correlate with the duration of surgery. There was no akinesia in the majority of cases with either method, but the surgeon was sometimes limited by the akinesia of the medial rectus muscle and often by that of the inferior rectus muscle with elevation of the globe. CONCLUSION: Greenbaum's method and Ripart's method are two subtenon anesthesic techniques characterized by an immediate, intense and prolonged analgesia (sometimes 60mn). Complete akinesia is rare and this is sometimes limiting. Chemosis was more often associated with Greenbaum's method, but Ripart's method carries the potential for needle-related complications. PMID- 11941238 TI - [Surgical indications for diabetic maculopathy]. AB - INTRODUCTION: The place of surgery in the treatment of diabetic macular edema is still not clearly defined even though the functional benefits of photocoagulation are less than satisfactory. Key words: Diabetic maculopathy, vitrectomy, internal limitant layer, serous detachment, hard exsudates. EQUIPMENT AND METHODS: We conducted a retrospective study on 40 consecutive cases of diabetic patients, each suffering from serious diabetic maculopathy and for whom photocoagulation would be either impossible to carry out or ineffective. Eighty percent of this study sample exhibited solid vitreomacular adhesions at vitrectomy. The internal limitant layer was dissected systematically. In all cases, plugging by perfluorocarbon liquids during the operation helped posterior focal endophotocoagulation. A gas tamponade was used in all cases. In 18 cases, surgical extraction of large intra- and/or subretinal clumps of hard exudates was necessary to replace the posterior pole. RESULTS: The anatomical results were satisfactory in 97.5% of cases. The functional results were good but their interpretation is more difficult: the visual gain varied as a function of the clinical preoperative condition and how recent the condition was. The best results were obtained in edema with tractional predominance. The smallest visual gains were observed in cases of massive macular hard exudates. The most serious complication was a retinal detachment secondary to a parapapillary nasal retinal break occurring at a distance from the operation during a postoperative photocoagulation complement. DISCUSSION AND CONCLUSION: Vitrectomy released both tangential and axial tractional forces found in diabetic macular edema pathogenesis. The extraction of large clumps of exudates allowed us to replace serous retinal detachments and the photocoagulation of capillary anomalies. In addition, removing premacular vitreous body and gaseous plugging seemed to osmotically resorb the posterior pole edema. These surgical results have made us considerably reduce the use of photocoagulation for diffuse diabetic macular edema in the past four years. PMID- 11941239 TI - [Clinically isolated Morning glory syndrome]. AB - The Morning glory syndrome is an uncommon congenital optic disk anomaly. It is characterized by a funnel-shaped and enlarged dysplasic optic disk with white tissue, and retinal vessels arising from the periphery of the disk and running an abnormally straight course over the peripapillary retina. We report here the case of a 7-year-old girl with a delayed diagnosis of Morning glory syndrome discovered at a regular examination that showed amblyopia. The optic disk anomaly was isolated. The search for other associated morphological and/or functional abnormalities is mandatory (retinal and/or cerebral malformation, frontonasal dysplasia, endocrine irregularities, neurofibromatosis type 2). Periodic ophthalmological examinations are recommended due to the frequency of retinal detachment. PMID- 11941240 TI - [Corneal lesions caused by caterpillar hairs: four case studies]. AB - Caterpillar hairs disseminated by the wind can cause serious ocular problems in humans. We present 4 cases of corneal lesions caused by caterpillar hairs. Intense functional signs observed in exposed subjects should bring this diagnosis to mind. The severity of this affection is due to the possibility of intraocular migration, even many years after the first accident. Treatment consists of removing the caterpillar hair associated with an antibiotic and local anti inflammatory treatment. PMID- 11941241 TI - [New concepts in intraocular tumor therapeutics]. PMID- 11941242 TI - [New strategies in the management of retinoblastoma]. AB - It was rare that a child survived retinoblastoma at the beginning of the twentieth century. Today the survival rate is in the order of 95% in reference centers, with new strategies improving prognosis step by step. Systematic enucleation used to be the starting point of any true and structured management, until the advent of radiotherapy made it possible not only to save lives but also to retain some useful vision. Early diagnosis has enabled focal therapies such as photocoagulation, cryocoagulation, and radioactive applicators to open up a new era of targeted tumor treatment. However, the onset of nonocular tumors secondary to radiotherapy, the resistance of certain tumors to irradiation, and unsightly cosmetic consequences all justify research into alternative therapeutic strategies. New types of chemotherapy have shown spectacular results and are currently under study: chemoreduction to make large tumors more manageable and enable less aggressive treatment of tumors located in delicate sites, thermochemotherapy using the effect of heat on plasma membrane permeability to antimitotics, and chemotherapy associated with cyclosporine to reduce the multidrug resistance of certain tumors. The aim is to avoid primary enucleation and external beam radiation as far as possible. The future may lie in local chemotherapy, hyperthermia, and dynamic phototherapy, accelerated proton beam radiotherapy also has promising prospects. PMID- 11941244 TI - [Choroidal melanoma: current therapeutic approaches]. AB - The management of posterior uveal melanoma has evolved tremendously over the past decades and more recently there has been a trend toward more focal conservative treatment. Transpupillary thermotherapy (TTT) with infrared diode laser (810nm) is the newest modality used as primary treatment or as a complement to radiotherapy or surgical resection in selected cases of choroidal melanoma. Plaque radiotherapy or charged-particle irradiation is particularly recommended for medium- or small-sized uveal melanoma not suitable to TTT or resection. Special custom-designed plaque radiotherapy (iodine-125) can be used for the iris and ciliary body, or in juxtapaillary choroidal melanoma. The tumor control rate after plaque or charged-particle radiotherapy appears to be similar but charged particle irradiation may produce worse anterior-segment complications than plaque radiotherapy. Stereotactic radiation therapy for choroidal melanoma may be effective in controlling tumor growth, but the number of patients treated with this approach is too small to draw solid conclusions. Local tumor resection using trans-scleral resection is mainly suitable for selected iris, ciliary body, or anterior choroidal melanoma, particularly with smaller basal dimensions and greater thickness. Endoresection may preserve central vision or temporal field when radiotherapy would be expected to cause optic neuropathy. Longer follow-up is necessary to establish the efficacy of tumor control. Combined therapies (radiotherapy plus TTT or tumor resection plus TTT) appear to be more effective in decreasing the incidence of intraocular tumor recurrence. Enucleation is still performed for large uveal melanoma when there is no hope of regaining useful vision. Based on the published ophthalmology literature, it seems that enucleation carries the same survival prognosis as each of the conservative treatment modalities. PMID- 11941243 TI - [Ocular manifestations of cancer]. AB - Cancer may affect the eye and orbit as a direct result of metastatic neoplastic infiltration, compression, or circulating antibodies involving paraneoplastic retinal degeneration. A metastatic tumor to the uvea is the most common form of an intraocular metastatic process. The choroid is the most common site for uveal metastasis; metastases to the ciliary body, iris, retina, optic disk, and vitreous are rare. Approximately one-third of patients have no history of primary cancer at the time of ocular diagnosis. Breast and lung carcinomas for women and lung and gastrointestinal carcinomas for men most commonly metastasize to the eye and orbit. The short-term prognosis for vision is usually good after an individualized therapeutic approach (chemotherapy, hormonal therapy, external beam radiotherapy, or plaque radiotherapy), but the systemic prognosis is poor. The visual paraneoplastic syndromes encompass several distinct clinical and pathological entities including carcinoma-associated retinopathy (CAR), melanoma associated retinopathy (MAR), and bilateral diffuse melanocytic uveal proliferation (BDUMP). The CAR syndrome affects photoreceptors, MAR is thought to affect bipolar cell function, and BDUMP targets the uveal tract. Identification of circulating antibodies against retinal proteins (recovering, 23-kDa retinal protein; 46-kDa and 60-kDa retinal proteins) serves to recognize the paraneoplastic nature of the patient's symptoms, which frequently develop before the cancer is diagnosed. Anecdotal therapeutic responses are described after systemic steroids, immunoglobulin injection, and plasmapheresis. Recognition of their visual symptoms and ocular findings should alert the ophthalmologist to the possibility of cancer and systemic evaluation should be pursued. PMID- 11941245 TI - [Advantages of digital indocyanine green angiography for diagnosing choroidal tumors]. AB - Indocyanine green angiography (ICG) is a well-known ancillary test for evaluation of choroidal tumors. The authors review the different images seen in choroidal nevus, melanoma, metastasis, hemangioma, osteoma, and vortex vein varix. ICG appears to be useful for the diagnosis of non pigmented choroidal tumors. In addition, ICG provides good information on the limits of pigmented tumors, particularly for the follow-up of a growing tumor and the choice of treatment. PMID- 11941247 TI - [Lisch nodules]. PMID- 11941248 TI - [Basal cell carcinoma associated with a squamous cell papilloma at the inner part of the eyelid]. AB - There are numerous variants of cutaneous tumors involving the eyelids. Tumors of a different nature may at times be observed simultaneously in the same area of the eyelid. A clinicopathologic case of a 36-year-old male patient with 2 different cutaneous tumors at the nasal part of the left eyelid is reported. One was a nodular tumor on the inner canthus with a pearly appearance; the other had a papillomatous pattern. After surgical removal, the histopathological study of the tumors disclosed a typical basal cell carcinoma and a squamous cell papilloma. Both tumors can be commonly observed on the eyelids and surgical excision cured the patient. PMID- 11941249 TI - [Vitrectomy without scleral buckle versus ab-externo approach for pseudophakic retinal detachment: comparative retrospective study]. AB - PURPOSE: Retinal detachment (RD) is a major complication of cataract surgery, which can be treated by either primary vitrectomy without indentation or the scleral buckling procedure. The aim of this study is to compare the results of these two techniques for the treatment of pseudophakic RD. PATIENTS AND METHODS: The charts of 40 patients (40 eyes) treated with scleral buckling for a primary pseudophakic RD were retrospectively studied and compared to the charts of 32 patients (32 eyes) treated with primary vitrectomy without scleral buckle during the same period by the same surgeons. To obtain comparable samples, patients with giant retinal tears, vitreous hemorrhage, and severe preoperative proliferative vitreoretinopathy (PVR) were not included. Minimal follow-up was 6 months. RESULTS: The primary success rate was 84% in the vitrectomy group and 82.5% in the ab-externo group. Final anatomical success was observed in 100% of cases in the vitrectomy group and in 95% of cases in the ab-externo group. Final visual acuity was 0.5 or better in 44% of cases in the vitrectomy group and 37.5% in the ab-externo group. The duration of the surgery was significantly lower in the ab externo group, whereas the hospital stay tended to be lower in the vitrectomy group. In the vitrectomy group, postoperative PVR developed in 3 eyes and new or undetected breaks were responsible for failure of the initial procedure in 2 eyes. CONCLUSION: Primary vitrectomy appears to be as effective as scleral buckling procedures for the treatment of pseudophakic RD. PMID- 11941250 TI - [Three-dimensionnal representation and descriptive geometry of the pure spherical and pure cylindrical profiles of Excimer photoablation]. AB - PURPOSE: To analyze the theoretical shape of the lenticules of corneal tissue ablated using Excimer laser for the correction of pure negative and positive spherical and cylindrical refractive errors, and to investigate the possible correlations between their spatial configuration and the clinical outcome for each procedure. METHODS: To obtain a theoretical three-dimensional representation of each ablated lenticule, we used software that performs boolean operations on three-dimensional virtual surfaces (Bryce 3D, Metacreation, Dublin, Ireland). The representation of the theoretical shape of the refractive lenticule etched by a given profile of ablation based on a paraxial model over a circular optical zone was obtained by performing appropriate Boolean operations between different preoperative and postoperative surfaces or elements. These operations were repeated to obtain the representation of the theoretical additional lenticule corresponding to the volume of tissue ablated to blend any abrupt optical zone edges with a constant slope. RESULTS: The lenticule corresponding to the negative spherical treatment had its maximum thickness in its center and no thickness at its edge, thus inducing a natural blend with the peripheral untreated cornea. The lenticule corresponding to the positive spherical treatment had its maximal thickness at the junction between the optical and transition zones, which are both circular. The lenticule corresponding to the negative cylindrical treatment had its maximal thickness along the flatter initial meridian and an elliptical transition zone contour. The lenticule corresponding to the positive cylindrical treatment had its maximal thickness at the edge of the optical zone along the initial flatter meridian, and no thickness at the perpendicular principal meridian. The transition zone is also elliptical. CONCLUSION: The basic characteristics of the lenticule corresponding to the pure negative spherical treatment could explain the lesser degree of postoperative refractive regression than that one occurring after pure positive spherical treatment and pure positive and negative cylindrical treatments. The characteristics of the lenticules corresponding to the negative and positive cylindrical treatments may also account for the substantial spherocylindrical coupling and regression, respectively commonly observed after such treatments. The three-dimensional representation of the lenticules ablated for the correction of pure spherical and cylindrical refractive errors may help to better understand the outcome of these procedures aimed at reshaping the anterior surface of the cornea. PMID- 11941251 TI - [Direct heterochromatic comparison for color perception evaluation]. AB - We analyzed the data obtained from the Malbrel chromatometer, a device that examines color vision. The principle of the chromatometer is based on the opponent chromatic receptive fields of the retina. It shows two adjacent luminous windows of small size that can be either red and green or yellow and blue. The patient's task is to adjust the brightness of the red window (or the yellow window) to a fixed green (or blue) window luminance. Several luminance levels were studied and data were recorded using specific software. The luminance and the chromatic coordinates of the windows have been determined. The aim of the statistical study was to compare the data with a simple distribution law and therefore to know if the chromatometre allows measurement of color vision. In this investigation, 95 eyes were studied. Each patient has to match 4 red/green brightness levels and 4 yellow/blue brightness levels. The main characteristics were evaluated: the means, medians and standard deviations. Then, from the qq plots we verified that the data distribution was near normal distribution. The percentile curves allowed 3 types of color vision to be determined. Around the median normal color vision can be defined. Between the 10(th) and the 20(th) percentiles and between the 80(th) and the 90(th) percentiles, a small anomaly was present. Below the 10(th) percentile or above the 90(th) percentile, a substantial anomaly was present. PMID- 11941252 TI - [Silent sinus syndrome: a rare case of enophthalmia]. AB - INTRODUCTION: The causes of spontaneous enophthalmos, without trauma, surgical factors, or systemic illness, which can be found are rare. A particular entity called silent sinus syndrome in the Anglo-Saxon literature, which concerns the long-term effect of hypoventilation of the maxillary sinus responsible for an attraction of the sinus walls, resulting in sagging of the orbital floor, itself responsible for the enophthalmos. CASE REPORT: We report the case about a 29-year old woman, presenting a left-side spontaneous enophthalmos, with sinus-type pains for 2 months. The clinical examination revealed a horizontal diplopia when looking leftwards. Computed tomography showed a maxillary homolateral sinusitis with characteristic attraction of the sinus walls. The draining of the sinus collection by inferior endoscopic enlargement of the maxillary ostium led to pain resolution. The enophthalmos and diplopia were stabilized. CONCLUSION: After eradicating the orbital causes of enophthalmos, the sinus causes were sought more precisely, the pauci-symptomatic causes such as the silent sinus syndrome. The recognition of this syndrome stopped the progression of enophthalmos by making the maxillary sinus permeable again. A surgical reconstruction of the orbital floor can be proposed in order to correct the motility of the eyes and esthetic considerations. PMID- 11941253 TI - [Full optical correction after cycloplegia in headache]. AB - PURPOSE: To study the advantages of cycloplegia in the accurate assessment of refractive errors and to evaluate the impact of full optical correction in the course of headache. MATERIAL: and methods: A prospective study was carried out in 164 eyes of 82 patients with headache. Corrected visual acuity was 10/10 in all cases. Refraction was measured before and under cycloplegia by cyclopentolate with a CANON R 50 infrared autorefractometer. Full optical cycloplegic correction based on the value of refraction under cycloplegia was prescribed in all cases. Mean follow-up was 10 months. Statistical analyses were used: Student's t test, Pearson coefficient correlation, and Chi2. RESULTS: Under cycloplegia, hypermetropia was significantly the most frequent ametropia (67.1%, p<10(- 9)). The difference between the spherical equivalent, before and during cycloplegia, was highly significant (p<10(- 5)) at 0.79. Optical correction eliminated headaches in 76.5% of the cases. CONCLUSION: The authors conclude on the role of accommodation in headache onset and the need to continue cycloplegia until the age of 45. The importance of optical correction in alleviating headaches is underlined. PMID- 11941254 TI - [Corneal harvesting from donors over 85 years of age: cornea outcome after banking and grafting]. AB - PURPOSE: To study the suitability of corneas from very old donors for graft after organ culture and their clinical and endothelial outcomes in recipients after perforating keratoplasty. METHODS: We stored 419 corneas at 31 degrees C for 13.1 +/- 4 days (mean +/- SD) and then divided them according to donor age: group 1, donors under 85 years of age (n=3 3 0, 79%, 16-84 years old), and group 2, donors over the age of 85 (n=8 9, 21%, 85-100 years old). Endothelial density at the time of harvest and before and after organ culture, rates of suitability for grafting, and clinical and endothelial outcomes of the 196 keratoplasty procedures were compared in a prospective longitudinal study of the 2 groups, with a mean follow-up of 25 months. The corneas were grafted with no pre established policy on matching with the age of the receiver. Statistical analysis was carried out on SPSS 10.0: Chi(2), Student t test, and Kaplan Meier survival curves. RESULTS: The average age of the donors was 72.1 +/- 16.7 years. The macroscopic aspect of the corneas was judged to be of slightly lower quality in group 2. No statistically significant difference was found in overall suitability for transplantation (group 1, 45% vs group 2, 54%, p=0.17) but elimination for low endothelial density was more frequent in group 2 (67% vs 39%, p=0.001). Cell density at the beginning of organ culture was lower in very old corneas than in younger corneas (respectively, 2116 +/- 368 vs 2 311 +/- 360 cell/mm(2), p=0.002) but no difference was apparent at the end of organ culture (respectively, 2 011 +/- 285 vs 2 090 +/- 296, p=0.12) because very old corneas lost fewer cells than younger ones (respectively, 5.6% vs 10.0%, p=0.001). There was no correlation between donor/receiver age (r=0. 337) but group 1 corneas were slightly more frequently allotted to receivers with normal endothelium (p=0.019). During surgery, the two groups did not differ in terms of the macroscopic aspect of the grafts. In the 196 grafted patients, and without age-matching, overall graft survival (86% vs 79%, p=0.275), visual acuity, and endothelial density (1 194 +/- 469 vs 1098 +/- 545 cells/mm(2), p=0.387) did not differ at the completion of the study. DISCUSSION: The corneas from very old donors were macroscopically of poorer quality and had a lower cellular endothelial density at harvesting, but these differences disappeared after organ culture because of greater cell loss in corneas from younger donors. Selection by organ culture ensures that functional, anatomical, and cellular results are not influenced by very old donor age. CONCLUSION: Considering the aging population in countries with a high standard of living, the techniques available for selecting corneas based on endothelial quality, and the increasing need for corneal grafts, the very old age should not be deemed off-limits for corneal harvesting. PMID- 11941255 TI - [Results of proton beam irradiation for treatment of choroidal melanoma]. AB - PURPOSE: To evaluate the results of proton beam irradiation of choroidal melanomas on a large series of patients. PATIENTS AND METHODS: Retrospective analysis of a series of patients treated with proton beam irradiation between 1991 and December 1998. The data were analyzed to evaluate the local tumor control as well as the general progression and metastatic rate of the patients. Statistical analysis served to isolate risk factors for relapse or metastasis. RESULTS: We treated 1062 patients during the study period, with a median follow up of 38 months. Local control was obtained for 97.1% of the patients. Tumors anterior to the equator were at risk for relapse. The survival rate was 92% at 2 years and 78% at 5 years. 73.1% of the 1062 patients died from metastasis, 6.1% of living patients presented with metastatic disease. The risk factors for death were the initial diameter, the age of the patient, and large tumor volume at diagnosis. Metastasis were essentially hepatic (94.6%). Risk factors for metastasis were: a large tumor volume, a lesion anterior or straddling the equator and the age of the patient. Ocular complications may induce a visual loss of 0.1 and less in 47% of the patients, due to optic nerve head and macular ischemia. 6% of the patients required secondary enucleation due to local complications (neovascular glaucoma). CONCLUSION: Proton beam irradiation of choroidal melanoma allows good tumor control and eye retention. The survival prognosis is associated with the initial volume of the tumor. The functional results may be improved and new therapeutics are needed to treat metastatic disease. PMID- 11941256 TI - [Update of orbital rhabdomyosarcoma therapy in children]. AB - INTRODUCTION: Rhabdomyosarcoma is the most frequent primitive orbital malignant tumor in children. If the treatment is started as soon as possible after discovery of the disease, the vital prognosis is considerably better than otherwise. The goal of this paper is to present the new therapeutic protocol and to report our experience in this field. MATERIAL AND METHOD: During the past 35 years, 102 cases of orbital tumors were collected in children under 15 years of age: 5 cases of rhabdomyosarcoma were cared for in our department. At the time of tumor diagnosis, the age of our patients ranged from 3 weeks to 13 years. After a biopsy or excision biopsy, all our cases were treated by chemotherapy with or without radiotherapy. Medication was mostly vincristine, ifosfamide and actinomycine D. When the result of the treatment was not satisfactory, carboplatine and epirubicine, vincristine as well as ifosfamide were given. Radiotherapy was performed only in particular cases or in recurrences. CONCLUSION: Rhabdomyosarcoma is a highly malignant tumor. Although rare, it is the most frequent of malignant tumors in children. It is important to keep it in mind in order to perform a biopsy enabling quick diagnosis and treatment following the modern protocol giving the highest chances of survival to these patients: about 98% in 3 years. PMID- 11941257 TI - [Peripheral retinal neovascularization and sarcoidosis: two case reports]. AB - Peripheral retinal neovascularization occurs in approximately 10% of cases of sarcoidosis. Its pathogenesis is unknown, but it probably results from retinal ischemia and/or inflammation. In cases of peripheral retinal neovascularization associated with sarcoidosis, sickle cell disease should be considered, even if sarcoidosis is histologically proved: new vessels seen in sickle cell disease and sarcoidosis may have a very similar pattern. We present two cases with histologically proven sarcoidosis who developed peripheral neovascularization: the first one had no associated disease and new vessels were likely to be related to sarcoidosis; in the second case, hemoglobin electrophoresis revealed hemoglobin SC, and provided diagnosis of sickle cell hemoglobinopathy. PMID- 11941258 TI - [Severe ocular burns by calcium carbide in a speleologist: a case report]. AB - A case of severe ocular burns in an amateur speleologist is reported. The explosion of his acetylene lamp caused the projection of calcium carbide particles, which induced burning of the cornea and conjunctiva in both eyes. He slowly recovered in several months. The pathophysiology of the burns, linked to the in situ production of lime, and their management are discussed. PMID- 11941259 TI - [Congenital megalocornea]. AB - Congenital megalocornea is a bilateral enlargement of the cornea's diameter that can be associated with trabecula and/or iris dysgenesis. This condition predisposes the patient to glaucoma. The physiopathology is poorly understood and may be related to lens induction during embryological development of the anterior of the eye. X-linked genetic transmission is found in 50% of cases, autosomal transmission in 40%, and sporadic transmission in the remaining 10%. Diagnosis is based on a thorough examination of the child's eye, often with general anesthesia. Different clinical aspects can be observed: simple megalocornea without ocular or systemic anomalies, megalocornea with ocular and/or systemic anomalies as well as multiple malformation syndromes, dermatological diseases, skeletal diseases, and genetic or chromosomic diseases. A systematic general pediatric examination is therefore necessary. Congenital megalocornea must be considered, first of all, as a differential diagnosis of primary congenital glaucoma and all of its etiologies. Treatment of associated ocular diseases, the search for an association with systemic diseases, and follow-up for the rest of the patient's life are indispensable. PMID- 11941260 TI - [Ophthalmic manifestations of nasal obstruction]. AB - Ophthalmic manifestations of nasal obstruction can involve palpebral, orbital and ophthalmic disturbances. They can be the consequence of -Direct contact with the nasal obstruction because the orbital cavity drains partly in to the nasal fossae: rings, lipoptosis, blephaochalasis, fat protrusions, aggravation of exophthalmy in Grave's disease. -Oral breathing: there are modifications in the shape of the palpebral fissure (round eye) by stretching of the facial mask and modifications of the orbital rims (sad eye) due to a lack of naso sinusal expansion, often associated with malocclusion. -A biomechanical correlation between the dental occlusion and the orbital area because of the presence of the maxillary sinus. Every occlusal disorder has an influence on projections of the supra-and infraorbital rims. PMID- 11941261 TI - [Eyelid neurofibroma affecting a young woman]. AB - A 43-year-old female patient had an amelanotic single tumor thickening the upper right eyelid and observed since early childhood. Clinically, it appeared as a firm mass, without sharp margins, giving a thickened appearance to the superior eyelid. The ophthalmic clinical examination was normal but some subcutaneous nodules attested to the diffusion of the disease recognized as a limited form of neurofibromatosis 1. Although well tolerated, it gave rise to a cosmetic disturbance and the patient was willing to undergo an excision. A complete surgical excision was performed on the lesions and the pathological study disclosed a typical neurofibroma. Regular observation of the different cutaneous nodules was recommended. PMID- 11941262 TI - [Eyelid precancerous keratosis]. AB - A 25-year-old-male patient had one keratinized tumor on the inferior right eyelid, first observed 5 months before. Its size had been increasing for the last three weeks. The visual function was normal. Clinical examination disclosed a papillomatous lesion on the cutaneous middle part of the eyelid. A surgical excision was performed under local anesthesia. The histopathological study found a papillomatous epidermal proliferation with both structural and cytological atypias, concluding with a precancerous keratosis of the eyelid. PMID- 11941263 TI - [What to do in case of acquired ptosis]. PMID- 11941264 TI - Ertapenem (Invanz)--a new parenteral carbapenem. PMID- 11941265 TI - Desloratadine (Clarinex). PMID- 11941266 TI - Echinacea for prevention and treatment of upper respiratory infections. PMID- 11941267 TI - Bosentan (Tracleer) for pulmonary arterial hypertension. PMID- 11941268 TI - Role of the endothelium and blood stasis in the appearance of varicose veins. AB - Today, chronic venous insufficiency affects millions of people but the investigation of veins and of venous diseases is still very poor. Additionally, the mechanism of the occurrence of varicose veins is not understood. Blood stasis is often associated with these pathological situations and we propose that resulting ischemic conditions can trigger the endothelium to release inflammatory mediators and growth factors. On one hand, the inflammatory mediators will recruit and activate neutrophils, which then infiltrate the venous wall and damage components of the extracellular matrix. On the other hand, growth factors induce smooth muscle cell migration, proliferation and de-differentiation into the synthetic phenotype, all together leading to the formation of neointima. These processes, being repeated over time, would eventually lead to alterations of the venous wall as observed in varicose veins. Venotropic drugs are used to treat chronic venous insufficiency. They are able to increase venous tone and to decrease vein and capillary permeability but they are also able to protect the endothelial cells against ischemia. Indeed, they target complexes of the mitochondrial respiratory chain and maintain ATP production during hypoxia. Hence, the cells are resistant to ischemia and do not release inflammatory mediators and growth factors. These drugs should thus be able to prevent the alterations of the venous wall induced by blood stasis. PMID- 11941269 TI - Exclusion and diagnosis of deep vein thrombosis in outpatients by sequential noninvasive tools. AB - Phlebography is the reference gold standard for the diagnosis of deep vein thrombosis (DVT), but due to its invasive nature and associated side effects it has been replaced by compression ultrasonography (CUS). Patients suspected of DVT are subjected to leg vein CUS that actually confirms DVT in only 16 to 28% of outpatients in large prospective management studies. CUS has a high positive predictive value of more than 98% for proximal DVT but usually misses calf vein thrombosis. Its negative predictive value for proximal DVT is about 97-98%, on the basis of which repeated scanning at day 7 after a negative first CUS (serial CUS) in outpatients with a first suspicion of DVT is advocated. Serial ultrasonography is costly and can be simplified and improved by the addition of clinical score and D-dimer testing. The safe exclusion of DVT by a rapid sensitive D-dimer test in combination with clinical score and/or CUS requires a negative predictive value of >99%. The negative predictive value for DVT is determined by the sensitivity of the rapid ELISA D-dimer test and the prevalence of DVT in subgroups of outpatients suspected of the condition. The prevalence of DVT in outpatients with a low, moderate and high clinical score varies widely from 3-10%, 15-30% and >70%, respectively. The combination of a low clinical score (prevalence DVT 3-5%) and a negative rapid ELISA D-dimer alone test will have a very high negative predictive value of >99.9% to exclude DVT without the need of CUS testing. The combination of a negative CUS and a negative rapid ELISA D-dimer test safely excludes DVT in patients with suspected DVT irrespective of the clinical score. The combination of a negative CUS, a low clinical score and a positive ELISA D-dimer but <1000 ng/ml excludes DVT with a negative predictive value of >99% without the need to repeat CUS. Patients with a negative CUS, scan but a positive ELISA D-dimer, and a moderate or high clinical score are still at risk with a probability of DVT of 3-5% and 20-30%, respectively and are thus candidates for repeated ultrasound scanning. The rapid ELISA D-dimer first followed by risk-based no, single or repeated CUS will be the most cost-effective strategy. PMID- 11941270 TI - Findings of the Naftidrofuryl in Quality of Life (NIQOL) European study program. AB - BACKGROUND: We report in this paper the findings of a pooled analysis of 3 previously published studies undertaken in Germany, France and Belgium to assess the effects of naftidrofuryl on the quality of life of patients with intermittent claudication. METHODS: A total of 754 patients were randomised in the 3 studies, 709 of whom (358 naftidrofuryl, 351 placebo) were available for the primary intention-to-treat analysis. The primary outcome variable was the change in the disease-related limitation of the quality of life as measured by the CLAU-S questionnaire. This instrument which has been validated in an international study, comprises 47 questions covering 5 dimensions: "daily living", "pain", "social life", "disease specific anxiety" and "mood". RESULTS: A multivariate analysis of covariance adjusted for baseline values, study effect and first order study treatment interaction, demonstrated the global superiority of naftidrofuryl over placebo (p<0.001). A separate covariance analysis for the 5 dimensions showed highly significant differences for "daily living", "pain", "social life" and "mood" (all p<0.01). CONCLUSIONS: In conclusion, this pooled analysis has shown that naftidrofuryl can significantly improve the quality of life of patients with intermittent claudication. These findings, taken together with evidence from previous studies that it improves treadmill walking distances, suggest that naftidrofuryl can play a useful role in the treatment of this condition. PMID- 11941271 TI - The ArtAssist Device in chronic lower limb ischemia. A pilot study. AB - BACKGROUND: Patients with chronic critical limb ischemia following a failed bypass graft or with non-reconstructable distal disease diagnosed angiographically, have a very poor prognosis. This is a prospective pilot study to assess the influence of the ArtAssist Device on pedal blood flow and amputation rate. METHODS: Thirty-three legs in 25 patients were evaluated. Ten legs presented with rest pain, and 23 legs with tissue loss. Nine legs had previously undergone bypass surgery. RESULTS: At a mean follow-up of 3 months, 14 (42%) legs were amputated, and 19 (58%) were saved. Eleven of the amputated legs were in patients with chronic renal failure, a known risk factor. The amputation rate, excluding this group, was 13.6% (3/22). Toe pressures measured initially and after 3 months on the pump showed a significant improvement (p=0.03). Forty percent of patients presenting with rest pain improved, while 26% of foot ulcers healed on the pump. Mortality rate was 12%. CONCLUSIONS: The results from this prospective study are encouraging but need to be validated in a larger prospective randomized study. PMID- 11941272 TI - Outpatient treatment of severe peripheral ischemia with intravenous intermittent low-dose iloprost. An open pilot study. AB - BACKGROUND: Iloprost given in a standard dose regimen (0.5-2 ng/kg/min for 6 hours daily over 21-28 days) has proven to be effective and safe in hospitalized patients with critical limb ischemia. Major drawbacks of the standard regimen are the high frequency of side effects, the long duration of the daily infusion, and a hospital stay of 3 to 4 weeks. Recently, the efficacy of low doses of iloprost (25 mg/day) was demonstrated. This open pilot study was undertaken to identify a more practical and cost-effective regimen with less side effects. The feasibility, efficacy and safety of an individually adapted, intermittently applied low-dose iloprost regimen in an outpatient setting were evaluated. METHODS: Twenty-seven patients with severe peripheral ischemia in the limbs or part of the limb due to various etiologies, who were eligible for outpatient treatment, were enrolled into the study. The infusion of iloprost (50 microg in 250 ml 0.9% saline) was started at 0.5 ng/kg BW/min and titrated to the individual optimum dose, which was defined as the maximum dose at which the patient felt entirely comfortable. The frequency of the iloprost infusions and the duration of the treatment were individually determined in each patient according to the severity of the clinical condition. Outcome endpoints were the response rates achieved by day 28, defined as substantial relief from rest pain and evidence of ulcer healing. The patients were followed up for a minimum of 6 months. RESULTS: A total of 27 patients (15 male, 12 female, mean age 65 years) were treated. Twenty-four patients received daily infusions with a break at weekends (5 times/week); 3 patients were treated every second day (3 times a week). The mean daily iloprost dose actually given was 20+/-5 microg, the mean duration of treatment was 3.6+/-0.8 weeks, i.e. a mean of 17+/-4 infusions were administered. Six patients with one-vessel run-off underwent percutaneous transluminal angioplasty (PTA) of their single calf vessel. Twenty-six patients showed clinical improvement by day 28; excluding those who had had PTA, the response rate to iloprost was 74% (20/27). No patient required admission to hospital while receiving outpatient treatment; no side effects occurred after adjustment to the optimum dose. At long-term follow-up (11+/-3 months), 76% of patients were alive and had a viable limb. CONCLUSIONS: In a limited number of patients with severe peripheral ischemia of various etiologies, long-term outpatient treatment with an individually adapted low-dose iloprost regimen was feasible and safe. Our data suggest that flexible treatment modalities might be as effective as rigid standard treatment regimens, the former being more advantageous in terms of greater practicability and cost-effectiveness due to outpatient management. Further studies are needed to confirm the efficacy of this individually adapted, low-dose outpatient iloprost treatment regimen in a larger number of patients. PMID- 11941273 TI - The effect of peripheral vascular disease on the serum levels of natural anti oxidants: bilirubin and albumin. AB - BACKGROUND: Bilirubin and albumin may act as antioxidants. Their circulating levels are lower in those patients with ischemic heart disease (IHD) and could be further reduced by more extensive atherosclerosis, i.e. peripheral vascular disease (PVD). METHODS: Serum bilirubin and albumin were measured in 456 patients classified into 3 groups: 1) no clinically evident cardiovascular disease (CVD), 2) IHD present and 3) PVD present. Smoking status and gender (which affect bilirubin and albumin levels in healthy individuals) were considered separately. RESULTS: Bilirubin was lower in smoking men without CVD or with PVD than in non smokers (p=0.02 and p=0.04, respectively) in the same groups. Non-smoking women without CVD had significantly (p=0.004) lower bilirubin levels than the corresponding group of men. Frequency analysis of male non-smokers revealed significantly (p=0.04) more patients with a lower bilirubin (<6.5 mmol/l) in the PVD compared with the no CVD group. Albumin levels showed the same trends. CONCLUSIONS: Prospective studies should consider smoking and gender when assessing the relevance of bilirubin and/or albumin levels in patients with vascular disease. Our findings support those of other studies that show that low serum bilirubin and albumin levels are associated with the presence of vascular disease. PMID- 11941274 TI - The V34L polymorphism of factor XIII and peripheral arterial disease. AB - BACKGROUND: Factor XIII catalyzes crosslinking of fibrin in the last steps of the coagulation process. A common polymorphism in the gene for factor XIII A subunit (F13A1 V34L) has been associated with a decreased risk for coronary artery disease, cerebrovascular disease, and deep venous thrombosis. METHODS: To analyze the role of this polymorphism in peripheral arterial disease (PAD) we performed a case-control study including 873 patients with documented PAD and a total of 523 controls without vascular disease. The F13A1 genotype was determined by an allele specific polymerase chain reaction. RESULTS: Genotype distribution and allele frequencies were not significantly different between patients (VV: 51.9%; VL: 40.7%; LL: 7.4%) and controls (VV: 54.7%; VL: 39.2%; LL: 6.1%). Mean age at onset of the disease was significantly higher in LL homozygous subjects than in VV homozygous subjects (67.3 versus 64.1 years, p=0.017). Heterozygous subjects had an intermediate age at onset (65.1 years), suggesting a gene-dose effect. The association of the L34 variant with onset of PAD remained significant after adjustment for other risk factors. The effect was stronger in men than in women. CONCLUSIONS: We conclude that the F13A1 V34L polymorphism was not associated with the presence of PAD in our study, but may be linked to a later onset of the disease. PMID- 11941275 TI - Accentuated oxidative stress following reperfusion injury in diabetic rats. AB - BACKGROUND: This study investigates whether diabetes mellitus accentuates hind limb ischemia-reperfusion injury. METHODS: Male Wistar rats rendered diabetic (n=40) following injection of streptozotocin were compared to non-diabetic control rats (n=30). Each group was divided into sham, 4 hrs of hind limb ischemia, 4 hrs of ischemia followed by 10, 30 or 60 min of reperfusion. Plasma concentrations of an end-product of lipid peroxidation [malondialdehyde (MDA)] and antioxidants (vitamins A and E) were measured together with the resting membrane potential (RMP) of the gastrocnemius muscle. RESULTS: Following reperfusion, the diabetic group showed greater and more persistent elevation of MDA and greater reduction of antioxidants. This was associated with reduction in the RMP only in the diabetic group. There was significant correlation between MDA level and the RMP in both groups of animals. CONCLUSIONS: These results indicate that oxidative stress following reperfusion injury is greater in the presence of diabetes mellitus. This may lead to a decrease in the RMP and increase in the vascular permeability, which may be associated with more complications. PMID- 11941276 TI - Atherosclerosis of carotid arteries and the ace insertion/deletion polymorphism in subjects with diabetes mellitus type 2. AB - BACKGROUND: The aim of the present study was to investigate the association of the angiotensin-converting enzyme (ACE) insertion/deletion (I/D) polymorphism with the ultrasonographically evaluated severity and characteristics of carotid artery atherosclerosis in subjects with diabetes mellitus type 2. METHODS: We assessed 184 subjects with diabetes mellitus type 2, 75 males and 109 females, mean age 61.4+/-7.7 years. All subjects were receiving oral antidiabetic drugs for glycemic control and were free of cardiovascular events. The ACE genotype was analyzed by the polymerase chain reaction (PCR) technique. The ultrasonographic examination of the carotid arteries was performed in both B-mode imaging and Doppler ultrasound. The common carotid artery intima-media thickness was assessed 15-20 mm proximal to the dilatation of the carotid bulb. The atheromatous lesions were classified according to their echogenic characteristics as predominantly echolucent, mixed and predominantly echogenic with under 30, 30-70 and over 70% of the total plaque area echogenicity, respectively. RESULTS: From the total cohort 29 (15.8%) subjects had the II, 86 (46.7%) the ID and 69 (37.5%) the DD ACE genotypes. The mean carotid artery diameter stenosis was 37+/-17%, 43+/-19% and 40+/-20% (p=NS) and the intima media thickness was 0.94+/-0.24 mm, 0.97+/ 0.20 mm and 0.98+/-0.20 mm (p=NS) in the II, ID and DD subgroups, respectively. When the echogenicity was analyzed according to the ACE I/D polymorphism, 12 subjects (41.4%), 13 (44.8%) and 4 (13.8%) with II genotype had predominantly echogenic, mixed and predominantly echolucent lesions, respectively. The ID genotype diabetics were found to have predominantly echogenic plaques in 41 cases (47.7%), mixed in 30 (34.9%) and predominantly echolucent in 15 cases (17.4%). From the 69 DD subjects 19 (27.5%) had predominantly echogenic plaques, 26 (37.7%) had mixed and 24 (34.8%) had predominantly echolucent lesions. Predominantly echolucent plaques were more frequently encountered among diabetics with the DD genotype (p<0.05), even after correction for demographic characteristics, the main risk factors of atherosclerosis and blood glucose control. CONCLUSIONS: The ACE genotype seems to be associated with the echogenicity of carotid artery atheromatosis but not with the common carotid artery intima media thickness or the degree of internal carotid artery stenosis in subjects with type 2 diabetes mellitus. The DD genotype may be implicated in the increased cardiovascular risk that characterizes echolucent plaques. PMID- 11941277 TI - Dependence of morphological changes of the carotid arteries on essential hypertension and accompanying risk factors. AB - BACKGROUND: AIMS: to evaluate morphological changes (intima-media thickness, IMT) of the carotid arteries in patients being treated for essential hypertension (EH), and to discover whether this abnormality can be detected in normotensive offspring of subjects with EH (familial trait, FT); and to investigate the interrelationship between IMT and accompanying risk factors. METHODS: EXPERIMENTAL DESIGN: cross-sectional study. SETTING: angiology department, university teaching hospital. SUBJECTS: the study encompassed 172 subjects, of whom 46 were treated hypertonics aged 40-55 (49) years, and 44 age matched, normotensive volunteers as controls. We also investigated 41 normotensives with FT for essential hypertension aged 20-30 (25) years and 41 age- and sex-matched controls without FT. INTERVENTIONS: the hypertensive subjects were being treated either with long-acting calcium-channel antagonists or ACE-inhibitors. MEASURES: using high resolution ultrasound, IMT of the carotid bifurcation and of the common carotid artery was measured. RESULTS: In the hypertensives, the mean IMT was greater than that in the controls (0.92 (0.10) mm vs 0.72 (0.07) mm; p<0.00005). The IMT was independently related to accompanying risk factors: a positive family history of hypertension, age of the patient, duration of EH and the level of systolic/diastolic blood pressure (BP), body mass index and total/LDL-cholesterol. In subjects with FT, IMT was also greater compared to the control group (0.60 (0.05) mm vs 0.55 (0.04) mm; p<0.00005). IMT was not related to BP values. CONCLUSIONS: In treated essential patients with the EH, the IMT was increased. Individuals with FT also had greater IMT in the absence of elevated BP. The IMT in hypertensives was related to accompanying risk factors, which could be pathogenetic determinants of EH and/or its complications. PMID- 11941278 TI - The use of low molecular weight heparins for the prevention of postoperative venous thromboembolism in general surgery. A survey of practice in the United States. AB - BACKGROUND: Even though low molecular weight heparins (LMWHs) have become the standard for venous thromboembolism (VTE) prophylaxis in most European countries and Canada, it was not until recently that LMWHs were approved for use in the United States. The main objective of this study was to assess the current preferences and attitudes of United States surgeons toward the prevention of VTE with particular reference to LMWH. METHODS: A survey with questions relative to VTE awareness, risk factors, and prevention practices was mailed to 10,000 Fellows of the American College of Surgeons. RESULTS: A total of 1,145 (11.45%) usable questionnaires were returned. The vast majority (96%) of respondents use prophylaxis against VTE. Although LMWHs were rated first regarding efficacy and second regarding simplicity of use, conventional unfractionated heparin at fixed doses remains the preferred pharmacological agent for VTE prevention (74%), followed by 2 LMWHs: enoxaparin (34%) and dalteparin (16%). Overall, 52% of surgeons preferred physical methods over pharmacological methods when used separately and 26% of surgeons utilize combined physical-pharmacological modalities. CONCLUSIONS: North American general surgeons have substantially modified their approach to VTE prevention in the last 4 years. Physical methods and unfractionated heparin remain the preferred prophylactic modalities, but LMWHs have gained rapid acceptance since their approval for use for VTE prevention in North America. Even though the results of this survey must be interpreted with caution because of the limited response rate and possible sampling bias, they still reflect the current preferences and attitudes of North American surgeons toward prophylaxis. PMID- 11941279 TI - Chronic venous insufficiency. A common health problem in general practice in Greece. AB - BACKGROUND: A study was carried out in Greece with the aim of assessing the prevalence of chronic venous insufficiency (CVI) in Greece and to discuss the role of general practice in the epidemiology, early diagnosis, and initial management of this disease. METHODS: DESIGN: a prevalence study was carried out in rural Greece between January 17 and December 30, 1997. SETTING: 13 GPs, 11 of those working in 13 rural primary health centers, 1 in the University Hospital of Heraklion and 1 private sector from different Greek districts were invited to participate in the study. PARTICIPANTS AND METHODS: patients with the following criteria were entered into the study: weight, pain, cramps, burning, itching, formication and swelling. Complete information about patients' history was collected by the general practitioners (GPs) who used a semi-structured questionnaire. The diagnosis of CVI was established with Doppler ultrasound. RESULTS: Data were based on 6,119 questionnaires and GPs observed 4,502 randomly selected patients. The diagnosis of CVI was established in 993 patients and its prevalence was found to be 11.9% in males and 39.8% in females. Varicose veins were the most frequent clinical finding in patients with vein reflux and diabetes mellitus was the most common co-existing disease in males and hypertension in females. CONCLUSIONS: The prevalence of venous insufficiency (VI) appears to be a frequent health problem in general practice in Greece. It requires further investigation in order to explore the observed differences between various groups. PMID- 11941280 TI - Acute superficial venous thrombophlebitis: does emergency surgery have a role? AB - BACKGROUND: Superficial venous thrombophlebitis (SVT) of the long saphenous vein (LSV) has been shown to be associated with thrombus propagation into the common femoral vein in up to 44% of cases. Conservative management can thus result in deep vein thrombosis (DVT), deep vein insufficiency or fatal pulmonary embolism (PE). To examine the effects of emergency division of the sapheno-femoral junction (SFJ) on the deep venous system in SVT of the LSV we used pre- and postoperative venous duplex ultrasound. METHODS: Emergency division of the SFJ was performed in 17 patients presenting with acute superficial venous thrombophlebitis. All patients had duplex ultrasound, which demonstrated thrombus of the above knee long saphenous vein together with a normal deep venous system. A follow-up duplex ultrasound scan was arranged on discharge and at 2 months. RESULTS: No patient had propagation of thrombus into the deep venous system or a PE. One patient developed a non-occlusive clot in the popliteal vein at 2 months follow-up. All patients were discharged at 48 hours. CONCLUSIONS: Using duplex ultrasound it has been shown that emergency division of the SFJ is a safe and effective way of preventing serious complications caused by thrombus in above knee LSV SVT. PMID- 11941281 TI - Vascular centers in Europe. Results of a panel discussion at the 14th Meeting of the European Chapter of the International Union of Angiology (Cologne, Germany, May 25, 2001). PMID- 11941282 TI - The importance of communication between physicians and patients. Special considerations in the era of endovascular therapy. AB - Most of the lawsuits in medical practice are the result of inadequate communication between doctors and patients and litigation has very little to do with errors. In the era of the technological development and the implementation of new techniques such as endovascular therapy, good communication between physicians and patients is essential. The purpose of this review is to elicit important considerations on challenging issues relating to the art of communication between physicians and patients and to provide information on what the vascular surgeon or interventionalist should bear in mind in obtaining an adequate informed consent from the patients. PMID- 11941283 TI - Left renal artery aneurysm with left accessory renal arteries. A case report. AB - A 57-year-old man was suspected of having a left renal artery aneurysm on enhanced computed tomography (CT) findings. Aortic angiography showed 3 left renal arteries with an aneurysm of the distal main renal artery. A 3 mm upper pole renal artery which crossed near the aneurysm was preserved. Dissection of the aneurysm was followed by arterial reconstruction with end-to-end anastomosis. PMID- 11941284 TI - Minimally invasive parathyroid surgery. AB - The standard bilateral neck exploration in primary hyperparathyroidism (HPTH) has been challenged in the recent years by the general trend toward less invasive surgery. The development of more reliable preoperative imaging techniques such as Sestamibi scanning and high definition ultrasonography coupled with improvements in intraoperative rapid assays of intact parathyroid hormone have allowed unilateral explorations in most patients with primary HPTH. This article reviews the currently available preoperative parathyroid localization studies as well as the currently used minimally invasive parathyroidectomy (MIP) techniques, such as open approaches, radioguided surgery and endoscopic procedures. While some techniques are more popular than others, careful selection of patients with primary HPTH has resulted in comparable cure rates to the standard bilateral parathyroid exploration. PMID- 11941285 TI - [Thoracoscopic esophagectomy for esophageal cancer. Personal experience]. AB - BACKGROUND: Esophageal surgery was recently modified by minimally-invasive approach. Personal experience with the thoracoscopic technique for esophagectomy in patients with early stage esophageal cancer is described. METHODS. From 1996 to 2000 at the Department of Thoracic Surgery of the University of Rome "La Sapienza", 10 patients, 7 male and 3 female, underwent video-thoracoscopic esophagectomy for esophageal cancer. Median age was 64 years (range 53-72). With the patient in left lateral decubitus 4 ports were positioned between the 4th and 8th intercostal space. The thoracic esophagus was mobilized in the entire length and circumference with the connective tissue and peri-esophageal nodal stations. A cervicotomy followed by a median laparotomy for tubulization of the stomach was performed. RESULTS: Nobody required conversion to thoracotomy. No complication or intraoperative death were observed. The median thoracic time was 110 minutes (range 55-165). No death within 30 days after discharge was recorded. One patient presented left vocal cord paralysis. In one case a recurrence in cervical anastomosis two months after the operation was observed. One patient died after 36 month for metastatic spread. Eight patients are alive with no evidence of disease, with median follow-up of 20 months. CONCLUSIONS: In our experience, the video-toracoscopic approach is a viable and safe option for the treatment of early stage esophageal cancer. Low incidence of complications and local recurrence should encourage a most frequent use of this procedure. PMID- 11941286 TI - [Surgical treatment of Hashimoto's thyroiditis. Personal experience]. AB - BACKGROUND: To report personal experience in the surgical treatment of Hashimoto's thyroiditis. METHODS. Eight patients (7 females, 1 male, mean age 48 years old) referred to our Institution with diagnosis of Hashimoto's thyroiditis have been studied. All patients were evaluated by determination of serum thyroid hormones and of anti-thyreoglobulin and anti-microsomal antibodies; ultrasound and scintigraphic scans of the gland were performed in all cases and a cytological examination of fine needle aspiration (FNAC) of the nodules was evaluated. Diagnosis of Hashimoto's thyroiditis was suspected on the basis of clinical and laboratory data and was confirmed by cytology and histology on surgical specimens. Preoperative FNAC showed a Hashimoto's thyroiditis with thyroid differentiated carcinoma in 3 cases (37.5%), Hashimoto's thyroiditis in 1 case and chronic thyroiditis in 4 cases (50%). Seven patients underwent surgery, while 1 patient received a medical treatment; we performed 6 total thyroidectomies and 1 hemithyroidectomy. Histology on surgical specimens confirmed the diagnosis of Hashimoto's thyroiditis in all cases; in 3 patients an associated papillary thyroid carcinoma was found. RESULTS: Postoperative mortality was absent; no major postoperative complications (laryngeal nerve paralysis or permanent hypocalcemia) were recorded. Only 2 mild transient hypocalcemias have been observed. CONCLUSIONS: Total thyroidectomy is the technique of choice in surgical treatment of Hashimoto's thyroiditis, a self immune pathology which involves the whole gland and has a high correlation with differentiated thyroid carcinoma (37.5%). Total thyroidectomy warrants a radical and definitive control of the disease, without risk of relapse, with a low incidence of major complications, in experienced hands, and anyway lower than the morbility due to reinterventions. PMID- 11941287 TI - Biliary leaks after laparoscopic cholecystectomy. Results of the endoscopic treatment. AB - BACKGROUND: Significant postoperative bile leaks occur in approximately 0.8 to 1.1% of patients. The goal of endoscopic therapy is to eliminate the transpapillary pressure gradient, thereby permitting preferential transpapillary bile flow rather than extravasation at the site of leak. METHODS. Sixty-four patients were retrospectively evaluated. Endoscopic treatment comprised endoscopic sphincterotomy followed by insertion of a naso-biliary drainage or a stent. Retained stones were extracted by standard procedures. RESULTS: The cystic duct remnant was the site of bile extravasation in 50 cases, ducts of Luschka were the source in 4 cases, common bile duct in 6 cases and common hepatic duct in 4 cases. Retained bile duct stones were detected in 21 cases and papillary stenosis in 4 cases. Endoscopic therapy involved sphincterotomy in 25 cases with stones extraction in 21 cases followed by nasobiliary drain insertion, and placement of stent in the remainder. Bile leaks resolved in 96.9% of patients, on average 3 days in cases of associated stones or papillary stenosis, and 6.5 days in the remainder. Two cases of mild pancreatitis were evidenced from endoscopic treatment. CONCLUSIONS: Endoscopic management is the treatment of choice for postcholecystectomy bile leaks. PMID- 11941288 TI - Axillary lymphectomy in breast cancer. AB - BACKGROUND: The aim of the study was to demonstrate the prognostic value of sentinel node biopsy compared to the sampling of clinically suspected nodes and lymphectomy of the 3 axillary levels. METHODS. From October 1996 to January 1999, 60 patients with breast cancer with a diameter of 4 cm or under using different procedures of axillary lymphadenectomy. Sentinel node biopsy was performed using Giuliano's technique, followed by lymph nodes larger than 5 cm (lymph node sampling) and lastly all axillary lymph nodes (axillary lymphectomy at 3 levels). RESULTS: Sentinel nodes were identified in all patients and a mean of 3 sentinel nodes (range 1-5) were removed during the procedure. Histological analysis showed metastatic sentinel nodes in 21 cases. Lymph node sampling was possible in 43 patients who presented enlarged nodes. The mean number of lymph nodes removed was 6 (range 3-10). Lymph node metastasis was found in 10 patients and of these 7 had a metastatic sentinel node, whereas 3 had presented negative results. Histological tests in all 60 cases of complete axillary lymphectomy showed positive results in 4 cases confirming metastasis present in sentinel nodes. CONCLUSIONS: The results show that the association of lymph node sampling can improve the efficacy of sentinel node dissection, highlighting the rare cases of false negatives. In our study, total axillary lymphectomy did not add any information to the N parameter and was resolutive in a small percentage of cases. PMID- 11941290 TI - [CT pulmonary density mapping: surgical utility]. AB - BACKGROUND: The present paper considers the technique of CT scan maps of pulmonary isodensity, examining lung density differences as a function of the type of disease and considering their significance for the purposes of refined, useful diagnosis in a surgical context. METHODS. The method is used to examine 3 groups of subjects selected on a clinical/anamnestic basis and a further group already admitted for surgery. For each patient we obtained 2 thoracic density scans during the phase of maximum inspiration and expiration. On each scan we constructed 50 isodensity maps, the equivalent of more than 2500 measurements: the preliminary standard was represented by 100 wide windows to produce total "illumination" of the pulmonary fields. The isodensity windows were then codified differently. Subsequently, the density scans were analysed with the technique of scalar decomposition. RESULTS: The CT scan maps of lung isodensity proved useful for certain lung diseases in which early diagnosis, topographic extent of the pathology and the refined definition of the pathological picture provide important solutions as regards the indication and planning of surgical treatment and for the evaluation of the operative risk and prognosis. CONCLUSIONS: We consider that the technique is rapidly performed, not complex and inexpensive and is able to supply detailed information on the lung parenchyma such as to be used not only as a routine technique but also in emergencies. PMID- 11941289 TI - Immediate and delayed two-stage post-mastectomy breast reconstruction with implants. Our experience of general surgeons. AB - BACKGROUND: Breast reconstruction after a mastectomy, today is considered the best choice to improve the quality of life for the patient. The aim of the procedure is to consider the aesthetic results obtained and the various involvements, comparing them with those already reported in literature. METHODS. There is no certain data of the number of breast reconstructions performed in Italy in the few years. In 1999, in Veneto, reconstruction surgery after total breast removal, represented only 16% of the operations. In our surgery Division, from 1996 to 2000, there were breast reconstructions performed, either immediately or delayed on 87 patients out of 118 after mastectomy. 73.7% none over the age of 60 (range 27-60). The breast removal technique used is the Madden, while, for the reconstruction we have put into effect the submuscle prosthetic implant; 63 patients were subject to follow-up treatment during the period of 10 to 36 months. RESULTS: Seventy-eight percent of the women have judged the final aesthetic results the same or better than expected. Based on reconstruction with prosthesis, is the preferred procedure, even with serious complications which do not always influence the final results. This preference is because of the reduced operation time and the psychological and physical benefits due to immediate restoral of the mammary volume and shape. CONCLUSIONS: Some problems (Garavaglia Law) have slowed down the diffusion in Italy of this type of treatment which is now receiving more and more request and approval by the women affected with breast cancer. PMID- 11941291 TI - Selection criteria and long-term results of surgery in symptomatic rectocele. AB - BACKGROUND: The aim of this study was to confirm some selection criteria for the transrectal repair of the anterior rectocele and to compare our surgical results with those reported in the literature. METHODS: From January 1992 to December 1999, 30 females (mean age 52.9 years, range 28-70 yrs) affected by anterior rectocele were prospectively evaluated with a standard questionnaire, clinical examination, proctosigmoidoscopy, colonic transit time, dynamic defecography, anal EMG, anal manometry. Then, they were submitted to transrectal repair of rectocele with anterior plication of the rectal muscular wall. Fourteen (46.6%) of them were also submitted to perineal levatorplasty. Patients were followed postoperatively (mean 25.7 months) with the same standard questionnaire, clinical examination, defecography, and manometry. Results were tested by Fisher's Exact text, Wilcoxon's test, and "t"-test. RESULTS: Rectal dyschezia, incomplete evacuation, digital help in defecating, mean stool frequency, and rectal bleeding significantly improved. After 3 months, 30% of patients had no complaints, 40% had only 1-2 episodes/month complaints, 13.3% had evacuation only using laxatives, and 16.6% were unchanged. Defecography showed a significant reduction of the rectocele in 70% of patients after 3 months. Manometric parameters were not significantly modified. Four (28.6%) out of 14 patients submitted to perineal levatorplasty complained of dyspareunia. CONCLUSIONS: Our surgical results were comparable with those reported in the literature, with more than 80% of successful outcome. Preoperative clinical data and defecography were confirmed to be basic parameters in selecting patients for surgery. Colonic transit time, anal EMG, and anorectal manometry demonstrated to be useful to recognize conditions as slow colonic transit time, peripheral denervation, and reduced voluntary contraction that could lead to a less satisfactory outcome after surgery, and might benefit with a postoperative perineal rehabilitation by biofeedback and anal electrostimulations. The perineal levatorplasty is not suitable in young females, due to the risk of dyspareunia. PMID- 11941292 TI - Radiofrequency ablation of hepatocellular carcinoma. AB - The majority of patients with primary or metastatic hepatic tumors are not candidates for resection because of tumor size, location near major intrahepatic blood vessels precluding a margin-negative resection, multifocality, or inadequate hepatic function related to coexistent cirrhosis. Radiofrequency ablation (RFA) is an evolving technology being used to treat patients with unresectable primary and metastatic hepatic cancers. RFA produces coagulative necrosis of tumor through local tissue heating. Liver tumors are treated percutaneously, laparoscopically, or during laparotomy using ultrasonography to identify tumors and to guide placement of the RFA needle electrode. For tumors smaller than 2.0 cm in diameter, one or two deployments of the monopolar multiple array needle electrode is sufficient to produce complete coagulative necrosis of the tumor. However, with increasing size of the tumor, there is a concomitant increase in the number of deployments of the needle electrode and the overall time necessary to produce complete coagulative necrosis of the tumor. In general, RFA is a safe, well-tolerated, effective treatment for unresectable hepatic malignancies less than 6.0 cm in diameter. Effective treatment of larger tumors awaits the development of more powerful, larger array monopolar and bipolar RFA technologies. PMID- 11941293 TI - [The evolution of viscerosynthesis and its methodological implications: from manual sutures to biocompatible materials]. AB - This study aims to provide an overview of the different suturing techniques used in surgery, paying special attention to cancer surgery which, compared to general surgery, has precise requirements correlated to the type of patient and form of exeresis used, which often tends to be extensive. In this panorama which manages to avoid any form of superficiality, the discussion ranges from mechanical automatic suturing devices to recent advances in so-called biological repair, without omitting an analysis of the gold standard of tissue repair, namely suture thread, which appears to have been overwhelmed by the enthusiasm for technological innovation, even in the literature. PMID- 11941294 TI - [Biliary tract carcinoma. Case report]. AB - Carcinoma of the main biliary way is a condition that often leads to a fatal outcome owing to the rapid growth and late onset of the symptoms. The importance of a correct, timely diagnosis is stressed, with reference to a clinical case in which even CT scanning did not confirm either the presence of the neoformation or the dilation of the intra and extrahepatic bile ways. The usefulness of humoral markers for the diagnosis of the disease in its preclinical phase in the interests of surgical treatment that is as radical as possible is underlined. PMID- 11941295 TI - [Malignant carcinoid of the last ileal ansa. Report on 2 consecutive clinical cases]. AB - Carcinoid tumors are very rare neoplasms, arising from enterochromaffin cells, classified in Apud system, exhibiting an intermediate malignancy, because of their long lasting clinical silence and low evolution to advanced stage. At the same time, these features unfortunately cause a high incidence of lymphatic and liver metastases, visible at first diagnostic approach, which are also determined by aspecific symptoms and signs, especially involving jejunal and ileal carcinoids, as the two cases described, and by very frequent absence, in current clinical practice, of pathognomonic carcinoid syndrome. Two carcinoids located into the distal ileal ansa, strictly adjacent to the ileocecal valve, are reported; the first tumor, accompanied by lymphonodal positivity, the second by a solitary hepatic metastasis, requiring segmentectomy of the liver, in addition to right hemicolectomy. PMID- 11941296 TI - Giant diverticulum of the sigmoid colon with perforation. Report of a case. AB - We present a case of perforated giant diverticulum of the sigmoid colon. This condition is extremely rare and only a few cases have so far been reported in the literature. Our case involved a 55-year old woman. Diagnosis was easy with barium enema and CT scan examination. Laparotomy revealed a giant diverticulum of the sigmoid colon compressing adjacent structures with signs of inflammation. An en bloc resection of the sigmoid colon, ovary and fallopian tube was performed with primary colon-rectal anastomosis. The post-operative course was uneventful. PMID- 11941297 TI - [Primary non-Hodgkin's lymphoma of the colon: a rare but possible location. Therapeutic approach. Description of a clinical case and review of the literature]. AB - On the basis of a case of large cell, highly malignant, non-Hodgkin's lymphoma of the colon, the authors describe the special features of this rare location; the general aspects of this rare pathology of the colon are then considered. The clinical case relates to a large cell, highly malignant, non-Hodgkin's colic lymphoma located in the caecum at two thirds proximally of the ascending colon of which it occupies half the lumen. The patient had come to our observation for a quite aspecific symptomatology characterised by digestive disturbances and a tendency to stipsis. In disaccord with certain literature reports, surgical treatment was considered by the authors the key to the therapeutic approach. Surgical exeresis should be as radical as possible to permit complementary therapies (chemotherapy and radio-therapy) to act with most effectiveness and thus offer the patient a better life. PMID- 11941298 TI - [Clinical case: post-traumatic thoracic aorta rupture. Considerations on emergency room treatment]. AB - A clinical case of post-traumatic rupture of the thoracic aorta at isthmus and descending aorta levels with favourable outcome is reported. Early suspicion of a possible aortic lesion related to the trauma modality which was characterized by sudden deceleration, immediate performance of a CT scan of the thorax with and without contrast medium and careful monitoring of arterial pressure which showed hypotension at controls during the stay in the emergency and radiological rooms, permitted us to opt for conservative treatment of the lesion with satisfactory outcome. PMID- 11941299 TI - [Laparoscopical marsupialization of symptomatic lymphoceles following kidney trans-plantation]. AB - Personal experience in the laparoscopic treatment of a lymphocele following kidney transplantation and a review of the literature are presented. We have treated a symptomatic lymphocele, which occurred a month after renal transplantation. It compressed the iliac vessels and obstructed the urine flow. Physical examinations revealed it consisted of two chambers; it measured 12 x 8 x 6 cm. A US-guided puncture was performed and a drainage tube out in place. A continuous flow of lymphatic liquid derived, and therefore we decided on surgical intervention, which was conducted in laparoscopy. A puncture of blue through the drainage tube was performed; under US-guidance, we made an opening in the peritoneal wall and in the wall of the lymphocele. A cauterization of the edges was conducted. An immediate improvement in subjective and objective symptoms was achieved. A four-month follow-up showed minimal residual effusion, slowly disappearing. Literature data and clinical evolution of the patient show that the laparoscopic approach is to be considered the "gold standard" for the treatment of symptomatic lymphocele following kidney transplantation. The authors recommend the use of US-guidance to single out anatomic structures, particularly for surgeons not trained in this procedure. PMID- 11941301 TI - A review of telemedicine in vascular surgery. AB - Telemedicine refers to the delivery of medical care through telecommunications and has been utilized by many medical specialists. In its basic form, telemedicine can involve the use of a telephone or fax. More advanced forms are the transmission of still images, often referred to as "store-and-forward" technology, or real-time two-way interactive video. The former is possible over existing phone lines or the Internet and has enjoyed success in visually oriented disciplines such as radiology and dermatology. The latter requires high bandwidth communication lines and is therefore considerably more expensive. This article reviews the use of telemedicine as applied specifically to vascular surgery. Initial studies indicate that store-and-forward technology can be used to adequately assess and treat wounds in vascular patients. A recent study reported the use of two-way interactive video for remote assessment of a wide variety of vascular patients. Diagnosis and treatment recommendations by the remote physician were found to be comparable to conventional on-site examinations. Patient satisfaction with the telemedicine examination was noted to be extremely high. In conclusion, telemedicine, although not commonly used in vascular surgery, has the potential of increasing patient access to specialty care, while decreasing patient or physician travel. The use of telemedicine in vascular surgery will likely continue to expand as technology improves and costs decrease. PMID- 11941300 TI - [External oblique muscle-cutaneous flap use of the in the reconstruction of large chest wall defects after mastectomy]. AB - The use of an external oblique muscle-cutaneous flap in the reconstruction of large chest wall defects after mastectomy is described. The flap is drown as a V Y advancement-rotation flap, laterally based, on the ipsilateral abdominal wall. The flap extends from the posterior axillary line to the linea alba, vascular supply is provided by the musculo-cutaneous perforating arteries of the intercostal vessels. Mobilization starts medially including the anterior rectus sheath, decollement continues between the external and the internal oblique muscles as far as the posterior axillary line. The abdominal wall, after flap mobilization, is reinforced by the plication of the internal oblique sheath. The flap was used in 13 patients with major anterior chest-wall defects after mastectomy. In one patient a marginal skin necrosis of about 2 cm was observed. The flap described differs from other external oblique flaps already described in several technical innovations that allow to obtain better functional and esthetic results. PMID- 11941303 TI - Primary immunodeficiency disorders: antibody deficiency. AB - As a group, antibody deficiencies represent the most common types of primary immune deficiencies in human subjects. Often symptoms do not appear until the latter part of the first year of life, as passively acquired IgG from the mother decreases to below protective levels. As with the T-cell immune deficiencies, the spectrum of antibody deficiencies is broad, ranging from the most severe type of antibody deficiency with totally absent B cells and serum Igs to patients who have a selective antibody deficiency with normal serum Ig. In addition to the increased susceptibility to infections, a number of other disease processes (eg, autoimmunity and malignancies) can be involved in the clinical presentation. Fortunately, the availability of intravenous immune serum globulin has made the management of these patients more complete. Recently, molecular immunology has led to identification of the gene or genes involved in many of these antibody deficiencies. As discussed in this review, this has led to a better elucidation of the B-cell development and differentiation pathways and a more complete understanding of the pathogenesis of many of these antibody deficiencies. PMID- 11941304 TI - Heterotrimeric G protein signaling: role in asthma and allergic inflammation. AB - Asthma and rhinitis are pathophysiologic conditions associated with a prototypical allergic response to inhaled allergens consisting of both neuromechanical and inflammatory components. Heptahelical receptors that bind guanosine triphosphate-binding proteins (G proteins), referred to as G protein coupled receptors (GPCRs), have been intimately linked with asthma and allergic inflammation for many years. G protein signaling mediates responses throughout the immune, nervous, and muscular systems that might contribute to the pathogenesis of allergic processes and asthma. For example, GPCR agonists or antagonists are used as therapies for asthma either by promoting airway smooth muscle relaxation (beta2 adrenergic receptor agonists) or by inhibiting inflammation in the nasal mucosa and airways (cysteinyl leukotriene receptor antagonists). The focus of this review is to explore how downstream signaling cascades elicited by GPCR activation contribute to the allergic phenotype and the mechanism by which pharmaceuticals alter signaling to generate a therapeutic effect. We also discuss physiologic modulators of G protein signaling, such as regulator of G protein signaling proteins and G protein receptor kinases, inasmuch as they represent potential new therapeutic targets in the treatment of atopy and other inflammatory conditions. PMID- 11941305 TI - Air sampling in occupational asthma. PMID- 11941306 TI - Sinusitis and asthma: entering the realm of evidence-based medicine. PMID- 11941307 TI - Tissue-specific glucocorticoid resistance-hypersensitivity syndromes: multifactorial states of clinical importance. PMID- 11941309 TI - Exhaled leukotrienes and prostaglandins in asthma. AB - BACKGROUND: Most of the studies investigating the role of leukotrienes (LTs) and prostaglandins (PGs) in asthma have used invasive (eg, bronchoalveolar lavage fluid) or semi-invasive (eg, sputum induction) techniques. Others have measured eicosanoids in plasma or urine, probably reflecting systemic rather than lung inflammation. Collection of exhaled breath condensate (EBC) is a noninvasive method to collect airway secretions. OBJECTIVE: We sought to investigate whether eicosanoids are measurable in EBC, to show possible differences in their concentrations in asthmatic patients and healthy subjects, and to investigate whether exhaled eicosanoids correlate with exhaled nitric oxide (NO), a marker of airway inflammation. METHODS: Twelve healthy nonsmokers and 15 steroid-naive patients with mild asthma were studied. Subjects attended on one occasion for pulmonary function tests, collection of EBC, and exhaled NO measurements. Exhaled LTB(4)-like immunoreactivity, LTE(4)-like immunoreactivity, PGE(2)-like immunoreactivity, PGD(2)-methoxime, PGF(2)(alpha)-like immunoreactivity, and thromboxane B(2)-like immunoreactivity were measured by means of enzyme immunoassays. RESULTS: LTE(4)-like immunoreactivity and LTB(4)-like immunoreactivity were detectable in EBC in healthy subjects, and their levels in asthmatic patients were increased about 3-fold (P <.0001) and 2-fold (P <.0005), respectively. Exhaled NO was increased in asthmatic patients compared with healthy subjects (P <.0001). There was a correlation between exhaled LTB(4) and exhaled NO (r = 0.56, P <.04) in patients with asthma. When measurable, prostanoid levels were similar in asthmatic patients and control subjects. CONCLUSIONS: Exhaled LTE(4) and LTB(4) are increased in steroid-naive patients with mild asthma. EBC may be proved to be a novel method to monitor airway inflammation in asthma. PMID- 11941310 TI - Chronic sinusitis in severe asthma is related to sputum eosinophilia. AB - BACKGROUND: Chronic rhinosinusitis and asthma are conditions that frequently coexist, particularly in severe asthma. The precise mechanism of the relationship between upper and lower airway inflammation is still a matter of debate. We hypothesized that the extent of inflammation in the nasal mucosa is related to lung function and inflammation in the bronchial mucosa in patients with severe asthma. OBJECTIVE: We sought to investigate the relationship between sinonasal inflammation as assessed on computed tomography (CT) scanning, lung function, sputum eosinophilia, and nitric oxide (NO) in exhaled air in patients with severe asthma. METHODS: Eighty-nine nonsmoking outpatients with severe asthma (29 men and 60 women; mean age 45 years; age range, 18-74 years) were included in this study. CT scans were scored (0-30) by a blinded investigator using a validated method. Lung function, NO in exhaled air, and sputum eosinophils were measured by using standard procedures. RESULTS: CT scans showed abnormalities in 84% of patients. Extensive sinus disease (score 12-30) was found in 24% of patients. There was a significant positive correlation between CT scores and eosinophils in peripheral blood (R(s) = 0.46) and induced sputum (R(s) = 0.40) and level of exhaled NO (R(s) = 0.45, P <.01). CT scores were also positively related to functional residual capacity and inversely related to diffusion capacity, particularly in patients with adult-onset asthma (R(s) = 0.47 and R(s) = -0.53, respectively). CONCLUSIONS: The results of this study show a direct relationship between sinonasal mucosa thickness and bronchial inflammation in severe asthma, particularly in patients with adult-onset disease. Whether sinus disease directly affects the intensity of bronchial inflammation is still an unanswered question. PMID- 11941311 TI - Harmful effect of immunotherapy in children with combined snail and mite allergy. AB - BACKGROUND: With respect to allergy, the possibility of cross-reactivity between snail and mite is well recognized, and anecdotal reports suggesting that allergen immunotherapy with mite extract can worsen snail-induced allergy exist. OBJECTIVE: We describe the effect of immunotherapy in 4 children with snail-mite allergy. METHODS: Four children (1 boy and 3 girls; 9-13 years of age) had consistent clinical histories (mild immediate respiratory symptoms after ingestion) and positive skin reactions for allergy to snail. They also had mite induced asthma and were therefore prescribed subcutaneous specific immunotherapy and subsequently followed. RESULTS: Several months (8-25) after starting immunotherapy, all children experienced life-threatening reactions, anaphylaxis, and respiratory failure after inadvertent ingestion of snail. Skin reactivity to the fresh food increased in all patients. CONCLUSIONS: This observation confirms that in patients with combined mite-snail allergy, immunotherapy should be avoided. PMID- 11941312 TI - Association of beta2-adrenoreceptor polymorphisms with nocturnal cough among atopic subjects but not with atopy and nonspecific bronchial hyperresponsiveness. AB - BACKGROUND: The reports from in vitro studies that beta(2)-adre-nergic receptor (B2AR) polymorphisms are associated with agonist-promoted downregulation have evoked considerable research interest for the roles of these polymorphisms to the pathogenesis of asthma. OBJECTIVE: We sought to evaluate the association between asthma phenotypes and B2AR polymorphisms at 2 sites (Arg16 --> Gly16 and Gln27 - > Glu27) in the general population. METHODS: Four hundred forty unrelated Korean adults were randomly selected, and asthma phenotypes were determined with a questionnaire, immunoassay, skin prick testing, and methacholine bronchial provocation testing. Genotypes of B2AR polymorphisms were determined with PCR based methods. RESULTS: No significant association was found between B2AR alleles and haplotypes and total IgE levels, skin test responses to aeroallergens, and bronchial responsiveness to methacholine. Among the atopic subjects, however, numbers of both Arg16 alleles and Arg16-Gln27 haplotypes were negatively associated with nocturnal cough, and in contrast, Gly16-Gln27 was positively associated with it. CONCLUSION: B2AR polymorphisms may play an important role in the expression of nocturnal cough in atopic subjects but not in the expression of atopy and bronchial hyperresponsiveness in a general population. PMID- 11941313 TI - Intranasal steroids and the risk of emergency department visits for asthma. AB - BACKGROUND: In patients with asthma, treatment for associated conditions, such as rhinitis, is recommended. It is unknown whether this treatment can reduce the risk for emergency department (ED) visits for asthma. OBJECTIVES: We sought to determine whether treatment with intranasal steroids or prescription antihistamines in persons with asthma is associated with a reduced risk for ED visits caused by asthma. METHODS: We performed a retrospective cohort study of members of a managed care organization aged greater than 5 years who were identified during the period of October 1991 to September 1994 as having a diagnosis of asthma by using a computerized medical record system. The main outcome measure was an ED visit for asthma. RESULTS: Of the 13,844 eligible persons, 1031 (7.4%) had an ED visit for asthma. The overall relative risk (RR) for an ED visit among those who received intranasal corticosteroids, adjusted for age, sex, frequency of orally inhaled corticosteroid and beta-agonist dispensing, amount and type of ambulatory care for asthma, and diagnosis of an upper airways condition (rhinitis, sinusitis, or otitis media), was 0.7 (95% confidence interval [CI], 0.59-0.94). For those receiving prescription antihistamines, the risk was indeterminate (RR, 0.9; 95% CI, 0.78-1.11). When different rates of dispensing for intranasal steroids were examined, a reduced risk was seen in ED visits in those with greater than 0 to 1 (RR, 0.7; 95% CI, 0.57-0.99) and greater than 3 (RR, 0.5; 95% CI, 0.23-1.05) dispensed prescriptions per year. CONCLUSIONS: Treatment of nasal conditions, particularly with intranasal steroids, confers significant protection against exacerbations of asthma leading to ED visits for asthma. These results support the use of intranasal steroids by individuals with asthma and upper airways conditions. PMID- 11941314 TI - Allergic diseases, skin prick test responses, and IgE levels in North Karelia, Finland, and the Republic of Karelia, Russia. AB - BACKGROUND: There is growing evidence to show that atopic diseases are more common in Western Europe than in the former socialist countries of Eastern Europe. OBJECTIVE: The aim of this study was to assess whether a similar difference exists between the most eastern province of Finland and a neighboring western district of Russia. METHODS: A random sample of 25- to 54-year-old subjects was taken from the population registers in the North Karelia Province in eastern Finland and from the Pitkaranta district across the border in the western part of Russia. Participants filled out a questionnaire on atopic and allergic symptoms and participated in a clinical study, which included skin prick tests with 11 airborne allergens and IgE measurements. RESULTS: Self-reported hay fever, allergic eye symptoms, atopic eczema, and asthma were much more common in Finland than in Russia. In Finland 34.2% and in Russia 21.8% had at least one positive skin prick test reaction. In Finland 21.5% but in Russia only 15.8% had at least one elevated allergen-specific IgE value of the 5 values measured. From 6% to 47% of the differences in self-reported symptoms between the countries were explained by atopy, as measured by means of skin prick testing or specific IgE values. CONCLUSIONS: A major difference in clinical allergic diseases and signs of symptoms was observed between the 2 geographically adjacent areas. This suggests that the difference in clinical allergy and atopic disposition is related to the differences in lifestyle and environmental factors. PMID- 11941315 TI - p38 Mitogen-activated protein kinase-induced glucocorticoid receptor phosphorylation reduces its activity: role in steroid-insensitive asthma. AB - BACKGROUND: Although glucocorticoids are the most effective treatment for chronic inflammatory diseases, such as asthma, some patients show a poor response. IL-2 combined with IL-4 can alter glucocorticoid receptor (GR) ligand-binding affinity and modulate glucocorticoid function. OBJECTIVE: We sought to confirm the altered ligand-binding affinity in a distinct group of steroid-dependent asthmatic subjects and examine the mechanism by which IL-2 and IL-4 modify the ligand binding affinity of the GR. METHODS: We examined PBMCs from healthy subjects, subjects with mild asthma, and steroid-dependent subjects with severe asthma using dexamethasone-binding assays and Western blot analysis of GR and phosphorylated activated transcription factor 2 expression. GR phosphorylation was measured after orthophosphate labeling and immunoprecipitation and cytokine production by means of ELISA. RESULTS: GR ligand-binding affinity was reduced in the nucleus but not in the cytoplasm of steroid-dependent asthmatic subjects compared with that seen in healthy subjects (dissociation constant, 39.8 +/- 4.6 vs. 6.79 +/- 0.8 nmol/L). This difference in ligand-binding affinity could be mimicked by IL-2 and IL-4 cotreatment and was blocked by the p38 mitogen activated kinase (MAPK) inhibitor SB203580. Activation of p38 MAPK by IL-2 and IL 4, as shown by means of phosphorylation of activated transcription factor 2, resulted in GR phosphorylation and reduced dexamethasone repression of LPS stimulated GM-CSF release. p38 MAPK phosphorylation of CD2(+) T cells occurred on serine residues. The ability of dexamethasone to modulate IL-10 release was also inhibited by IL-2 and IL-4 cotreatment. These effects were also inhibited by SB203580. CONCLUSION: These data show that p38 MAPK inhibitors may have potential in reversing glucocorticoid insensitivity and reestablishing the beneficial effects of glucocorticoids in patients with severe asthma. PMID- 11941316 TI - Pathways of anaphylaxis in the mouse. AB - BACKGROUND: Although anaphylaxis is classically mediated by IgE, FcepsilonRI, mast cells, and histamine, several rodent studies suggest that an alternative pathway involving IgG, FcgammaRIII, macrophages and platelets, and platelet activating factor (PAF) may be more important in the anaphylactic response to antigen challenge. OBJECTIVES: We sought to determine the relative roles of the classical and alternative pathways of anaphylaxis in a mouse model characterized by mastocytosis and a high level of antigen-specific IgE antibody. METHODS: Wild type, IgE-deficient, FcepsilonRI-deficient, and mast cell-deficient mice were immunized with goat anti-mouse IgD antibody, which induces mastocytosis and a large IgE and IgG anti-goat IgG response, and then challenged 14 days later with antigen (goat IgG) or rat anti-mouse IgE mAb. Specific vasoactive mediators, cell types, Ig isotypes, or Ig receptors were blocked or eliminated before challenge in some experiments. The severity of anaphylaxis was gauged by changes in body temperature, physical activity, and mortality. RESULTS: Equal doses of antigen or anti-IgE mAb induced similar anaphylactic responses. Anti-IgE mAb-induced anaphylaxis was FcepsilonRI and mast cell dependent and mediated predominantly by histamine. In contrast, neither mast cells nor platelets appeared important for antigen-induced anaphylaxis, which was FcgammaRIII and macrophage dependent and mediated predominantly by PAF. CONCLUSIONS: Antigen-induced anaphylaxis in the mouse proceeds primarily through the IgG, FcgammaRIII, macrophage, and PAF pathway, even in an experimental model that is characterized by strong mast cell and IgE responses. The presence of FcgammaRIII on human macrophages makes it possible that the IgG, FcgammaRIII, macrophage, and PAF pathway also contributes to human anaphylaxis. PMID- 11941317 TI - Predominant expression of 950delCAG of IL-18R alpha chain cDNA is associated with reduced IFN-gamma production and high serum IgE levels in atopic Japanese children. AB - BACKGROUND: We previously reported that serum IgE levels were negatively correlated with the amount of IFN-gamma produced by phytohemagglutinin-stimulated or IL-12-stimulated PBMCs and that one of the mechanisms of the pathogenesis of atopy was the reduced IFN-gamma production, which led to upregulated IgE production. OBJECTIVE: IL-18 is also known to be a strong inducer of IFN-gamma production. However, it has not yet been determined whether IL-18 is associated with atopic disease. METHODS: We investigated the response to IL-18 or IL-12 stimulation and the sequence of IL-18 receptor (IL-18R) alpha chain cDNA in 41 nonatopic controls and 39 atopic patients. RESULTS: Serum IgE level was negatively correlated with IFN-gamma production by PBMCs stimulated with IL-18. The IL-18R alpha chain cDNA of atopic patients was sequenced. We identified a 3 base deletion of the IL-18R alpha chain cDNA (950delCAG ), which was generated by alternative splicing, as determined on the basis of genomic sequence data for the IL-18R alpha chain gene. PBMCs with the predominant expression of 950delCAG significantly showed the reduced IFN-gamma production after IL-18 stimulation. There was a significant difference in the expression pattern of the IL-18R alpha chain transcript between the atopic patients and the nonatopic controls. CONCLUSION: According to these results, the dominant expression of the 950delCAG transcript of IL-18R alpha chain cDNA, which was associated with reduced IFN gamma production by IL-18 stimulation and high serum IgE levels, is predisposition to some atopic diseases. PMID- 11941318 TI - Leukotriene D4 upregulates eosinophil adhesion via the cysteinyl leukotriene 1 receptor. AB - BACKGROUND: Eosinophils (EOS) are one of the cellular sources of cysteinyl leukotrienes (cysLTs) in allergic inflammation. There is evidence that cysLT(1)receptor antagonists possess anti-inflammatory properties in vivo in asthmatic airways. Although the exact mechanism of action remains unknown, cysLTs might regulate the cellular responses involved in allergic inflammation. OBJECTIVE: The present study was undertaken to examine whether LTD(4)modifies the adhesive property of EOS. METHODS: EOS were isolated from the blood of healthy subjects. Their adhesion to tissue culture plates or recombinant human (rh) adhesion proteins was then examined in the presence or absence of LTD(4). RESULTS: LTD(4)significantly augmented EOS adhesion to tissue culture plates (adhesion: 5.0% +/- 0.5% by medium control vs 9.1% +/- 1.2% by 1 micromol/L; P <.01; n = 10). The enhanced adhesion induced by LTD(4) was blocked by pranlukast, a cysLT(1) receptor antagonist, or an anti-beta(2) integrin antibody. Flow cytometry analysis revealed that LTD(4) significantly enhanced the expression of CD11b and CD18 on the EOS surface. Finally, LTD(4) augmented EOS adhesion to rh intercellular cell adhesion molecule 1 but not to rh vascular cell adhesion molecule 1 or fibronectin. CONCLUSIONS: The results suggest that LTD(4) directly upregulates the adhesive property of EOS via the cysLT(1) receptor and beta(2) integrin. LTD(4) generated from EOS or cells of some other type might contribute to the development of phenotypic change in airway EOS. PMID- 11941319 TI - Evidence for a role of Langerhans cell-derived IL-16 in atopic dermatitis. AB - BACKGROUND: The factors controlling infiltration of inflammatory cells into atopic dermatitis (AD) lesions remain to be fully explored. Recently, epidermal cells in lesional AD were reported to contain increased messenger (m)RNA levels of IL-16, a cytokine that induces chemotactic responses in CD4(+)T cells, monocytes, and eosinophils. OBJECTIVES: We sought to determine the expression of IL-16 in epidermal cells in normal skin and skin from AD lesions and to investigate whether Langerhans cell (LC)-derived IL-16 may contribute to the initiation of atopic eczema. METHODS: The cutaneous expression of IL-16 was investigated by in situ hybridization and immunohistochemistry. Expression of IL 16 was also investigated in freshly isolated LCs and in keratinocytes by intracellular cytokine staining, quantitative real-time RT-PCR, and ELISA. RESULTS: Low levels of IL-16 mRNA, but no stored IL-16 protein, were detected in keratinocytes and LCs isolated from normal skin. Synthesis, storage, and secretion of IL-16 could be induced in LCs, but not keratinocytes, by activation with phorbol ester and ionomycin. In normal skin (n = 10) neither keratinocytes nor LCs expressed IL-16. In contrast, IL-16 was contained in approximately 40% of CD1a(+)LCs in patients with active AD (n = 16). IL-16 expression in LCs in patients with AD correlated with the number of infiltrating CD4(+)cells (r =.72, P =.0017) and was completely downregulated parallel to the clinical response of AD lesions to topical treatment with FK506. CONCLUSION: LC-derived IL-16 may participate in the recruitment and activation of inflammatory cells in AD. PMID- 11941320 TI - Milk-induced urticaria is associated with the expansion of T cells expressing cutaneous lymphocyte antigen. AB - BACKGROUND: Two forms of IgE-mediated skin reactions, atopic dermatitis (AD) and urticaria, have been associated with milk allergy. The reason for these distinct reactions is poorly understood. T cells expressing cutaneous lymphocyte antigen (CLA), a unique skin-homing receptor, are known to play an important role in AD. In contrast, the role of lymphocytes in patients with milk-induced urticaria is unclear. OBJECTIVE: The expression of the skin-specific homing receptor CLA after in vitro milk protein-specific stimulation was investigated to determine whether T-lymphocyte homing to the skin plays a role in food-induced urticaria. METHODS: Fourteen patients with milk-induced urticaria but no evidence of AD were included in the study and compared with 6 children with milk-induced AD, 6 children with milk-induced gastrointestinal diseases, and 6 nonatopic and 6 atopic individuals without milk allergy. PBMCs were cultured in the presence or absence of caseins or tetanus toxoid. T-cell proliferation was determined, and T-cell phenotyping was performed by means of flow cytometry with anti-CD4, anti-CD8, and anti-CLA mAbs. RESULTS: After in vitro stimulation with caseins, PBMCs from patients with milk-induced urticaria and AD had a significantly greater percentage of CD4(+) T cells expressing CLA than patients with milk-induced gastrointestinal symptoms and atopic or nonatopic control subjects. After tetanus stimulation in vitro, no significant difference between the groups was observed. T cells from both patients with milk-induced urticaria and control subjects proliferated well in response to caseins and tetanus. CONCLUSION: Lymphocytes expressing CLA are selectively activated in patients with milk-induced urticaria and may play an important role in the pathogenesis of this disease. Expression of CLA is not unique to milk-induced inflammation in the skin of patients with AD and milk allergy. PMID- 11941321 TI - TH1/TH2 cytokines and inflammatory cells in skin biopsy specimens from patients with chronic idiopathic urticaria: comparison with the allergen-induced late phase cutaneous reaction. AB - BACKGROUND: Previous studies have demonstrated infiltration of eosinophils, neutrophils, monocytes, and T cells in the skin of patients with chronic idiopathic urticaria (CIU), suggesting a possible T(H)2-type cytokine pathology analogous to the allergen-induced skin late-phase reaction (LPR). OBJECTIVE: We sought to compare skin biopsy specimens from patients with CIU and the allergen induced skin LPR for mRNA(+) cells for IL-4, IL-5, and IFN-gamma and inflammatory cell infiltration. METHODS: Skin biopsy specimens were obtained from 13 patients with CIU (6 had positive results for FcepsilonRI autoantibodies), 6 nonatopic control subjects, and 6 atopic subjects (before and after cutaneous allergen challenge). Cryostat sections were processed for immunohistochemistry and in situ hybridization by using the (35)S-riboprobes. RESULTS: There were significant increases in the numbers of intradermal CD3(+) (P =.007), CD4(+) (P =.004), CD8(+) (P =.012), and CD25(+) (P =.018) T cells, as well as eosinophils (P =.02), neutrophils (P =.01), basophils (P =.004), and macro-phages (P =.0014) in patients with CIU compared with numbers in nonatopic control subjects. There were lower numbers of tryptase-positive mast cells (P =.048). In the epidermis of patients with CIU, but not in that of normal subjects or in allergen-challenged biopsy specimens, there were increased numbers of CD3(+) T cells (P =.039). The profile of inflammatory cell infiltration in the allergen-induced skin LPR was similar to that in patients with CIU. In patients with CIU there was a T(H)0 cytokine profile, with significant increases in IL-4 (P =.0029), IL-5 (P =.0025), and IFN-gamma (P =.037) mRNA(+) cells. As expected, in the skin LPR there was an increase in IL-4 (P =.0082) and IL-5 (P =.0051), but not IFN-gamma, mRNA(+) cells. There were no significant differences in either the numbers of inflammatory cells or the cytokine pattern between patients with and without autoantibody. CONCLUSION: The molecular immunopathology of CIU is that of an eosinophil and basophil cell-mediated hypersensitivity reaction. Thus it is similar to the allergic skin LPR but has a T(H)0 rather than a T(H)2 cytokine profile. PMID- 11941322 TI - Inhaled latex allergen (Hev b 1). AB - BACKGROUND: IgE-mediated responses to natural rubber latex allergens have become a major health problem among recognized risk groups. OBJECTIVE: The purpose of this investigation was to measure the amounts of Hevea brasiliensis latex allergen (Hev b 1) inhaled and deposited on surfaces when latex or vinyl gloves were worn and compare the results with the conventional measures (breathing zone samplers) of occupational exposure. METHODS: Hev b 1 exposure was measured by nasal sampling and breathing zone sampling. Latex allergen exposure was generated by having each subject don a pair of powdered latex examination gloves and continuing his or her normal daily activity for 30 minutes. By means of adhesive tape, surface dust samples were collected from the surfaces of gloves, the subject's hands, and work areas. Sampling was performed with subjects wearing no gloves, subjects wearing powdered vinyl gloves, subjects wearing powdered latex gloves, and nearby colleagues wearing latex gloves. All samples were assayed through use of the HALOgen assay (Inhalix, Sydney, Australia) with a Hev b 1 specific mAb. Particles transporting latex allergen were identified by a surrounding immunostain halo, and these were quantified and reported as total numbers of particles inhaled, airborne, or found on surface areas evaluated. RESULTS: Study subjects inhaled 26 times more allergen when powdered latex gloves were worn than under the "no glove" and powdered vinyl glove conditions. During the same period, Hev b 1 particle levels measured in the ambient air through use of the breathing zone sampler increased by 24-fold. The median numbers of particles carrying Hev b 1 allergen per square centimeter on the surface of the hands after the wearing of latex and vinyl gloves were 1964 and 5, respectively. Latex allergen was physically associated both with cornstarch granules and with larger dust particles having a darker, more irregular appearance. CONCLUSION: In a laboratory where gloves are worn for protection, the use of latex gloves resulted in a 26-fold increase in inhaled latex allergen over background levels measured while vinyl gloves were worn as controls. Low levels of latex exposure also occurred when vinyl gloves or no gloves were worn; the reasons for this are under investigation. PMID- 11941323 TI - Human milk-specific mucosal lymphocytes of the gastrointestinal tract display a TH2 cytokine profile. AB - BACKGROUND: A number of gastrointestinal disorders, including allergic eosinophilic gastroenteritis and food protein-induced enteropathy, have been associated with milk hypersensitivity. The immunologic reactions appear to involve T cells that are activated by specific food proteins. OBJECTIVE: The present study was performed to examine the cytokine profiles of milk-specific lymphocytes from the duodenal lamina propria from children with milk-induced gastrointestinal diseases. METHODS: Duodenal biopsy specimens obtained from 10 patients with allergic eosinophilic gastroenteritis, food protein-induced enteropathy, or both and 12 control subjects were mechanically minced and cultured with either mitogens (i.e., polyclonal T-cell expansion) or milk proteins (i.e., milkspecific T-cell expansion). By using flow cytometry, expanded T cells were phenotyped with anti-CD4, anti-CD8, anti-IL-4, anti-IL-5, and anti IFN-gamma mAbs. The milk specificity of the lines was evaluated by means of the lymphocyte proliferation assay. In addition, the release of T(H)1, T(H)2, and T(H)3 cytokines was determined after restimulation. RESULTS: In patients and control subjects polyclonal expansion of mucosal lymphocytes resulted in predominantly T(H)1 cells. Milk-specific mucosal T-cell lines could be established in 60% of the patients but in none of the control subjects. In contrast to the polyclonal expansion of T cells, the milk-specific expansion of mucosal T cells showed a clear T(H)2 cytokine profile. On restimulation with milk protein, these cells showed a high proliferative response. They released T(H)2 cytokines, predominately IL-13, but failed to release T(H)3 cytokines important in the development of oral tolerance. CONCLUSION: The release of T(H)2 cytokines after stimulation of milk-specific mucosal T cells may play a pathogenic role in the inflammatory changes seen in milk-induced gastrointestinal disorders. PMID- 11941324 TI - Localized heat urticaria associated with mast cell and eosinophil degranulation. PMID- 11941325 TI - Anaphylactic reaction to apple juice containing acerola: cross-reactivity to latex due to prohevein. PMID- 11941326 TI - Allergen removal from hard floors: assessment of a range of sweeping devices. PMID- 11941328 TI - Intramuscular versus subcutaneous injection of epinephrine in the treatment of anaphylaxis. PMID- 11941327 TI - IgE autoantibodies monitored in a patient with atopic dermatitis under cyclosporin A treatment reflect tissue damage. PMID- 11941329 TI - Pioneers and milestones. Clemens von Pirquet, MD (1874-1929). PMID- 11941331 TI - Budesonide inhalation suspension: a nebulized corticosteroid for persistent asthma. AB - Guidelines for managing asthma in pediatric patients published by the American Academy of Allergy, Asthma, and Immunology and the American Academy of Pediatrics recommend the use of inhaled corticosteroids for the management of persistent asthma in infants and young children. When these guidelines were published, pressurized metered-dose inhalers and dry-powder inhalers were the only delivery devices available for inhaled corticosteroids in the United States. These devices can be difficult for young children to use correctly. Furthermore, no inhaled corticosteroid was approved in the United States for the treatment of children younger than 4 years. Budesonide inhalation suspension (Pulmicort Respules; AstraZeneca LP, Wilmington, Del) was developed to meet the medication delivery needs of infants and young children with persistent asthma. Pulmicort Respules is the first inhaled corticosteroid approved for administration by means of a nebulizer and the only inhaled corticosteroid approved in the United States for infants as young as 12 months. Budesonide has been studied extensively worldwide. In the United States the tolerability and efficacy of budesonide inhalation suspension were confirmed in 3 placebo-controlled multicenter trials. These studies demonstrated that both once- and twice-daily dosing of budesonide inhalation suspension (0.25-1 mg) improved pulmonary function and ameliorated asthma symptoms in infants and young children with persistent asthma. Budesonide inhalation suspension was well tolerated, and the incidences of reported adverse events were similar among patients in the budesonide, placebo, and conventional asthma therapy groups. This article reviews the results of these studies, as well as the pharmacokinetics, pharmacodynamics, and clinical use of budesonide inhalation suspension. PMID- 11941332 TI - Functional magnetic system. PMID- 11941333 TI - Indexes of malocclusion. PMID- 11941334 TI - Evidence-based orthodontics and the Twin-block appliance. PMID- 11941335 TI - Evidence-based orthodontics. PMID- 11941336 TI - The art of moving teeth in the 21st century. PMID- 11941337 TI - What price progress? PMID- 11941338 TI - Three-dimensional assessment of the effects of extraction and nonextraction orthodontic treatment on the face. AB - The purpose of this prospective study was to compare the 3-dimensional (3-D) effects on the face of extraction and nonextraction orthodontic treatment in patients with skeletal Class I patterns. The sample of 24 patients included 12 whose treatment included extractions and 12 who were treated without extractions. Pretreatment study casts were assessed to evaluate dental differences between the 2 groups. Pretreatment and posttreatment lateral cephalograms and optical surface scans were also compared. In the lateral cephalometric assessment, the only significant (P <.05) pretreatment difference was lower lip length. Posttreatment, the nasolabial angle and lower lip thickness were significantly different (P <.05). Registration of the average 3-D optical surface scans indicated that, before treatment, the nonextraction group had longer and broader faces by 5 to 7 mm, and the upper lip and labiomental fold were 3 to 5 mm farther forward than in the extraction group. Posttreatment, the nonextraction group still had larger faces, but the difference was smaller. Faces in the extraction group became relatively more protrusive with treatment. The surface shape analysis technique showed that the cheeks were flatter in the nonextraction group at the start of treatment, but this reversed with time. In the extraction group, the concavity of the labiomental fold increased, while the nonextraction group showed no change in this area. The study demonstrates that 3-D optical surface scans offer more data for analysis compared with lateral cephalograms alone. It also highlights the changes that can be detected with surface shape analysis. PMID- 11941339 TI - Numerical experiments on long-time orthodontic tooth movement. AB - In orthodontic treatment, teeth are moved by the use of specific force systems. The force system used depends on the patient's orthodontic situation characterized by the geometry of the tooth and the surrounding alveolar bone, which defines the position of the center of resistance. Therefore, the simulation of bone remodeling could be helpful for the treatment strategy. In this study, the optimal force system for bodily movement of a single-root tooth, with an orthodontic bracket attached, was determined. This was achieved by the use of the numerical finite element method, including a distinct mechanical bone-remodeling algorithm. This algorithm works with equilibrium iterations separated in 2 calculation steps. Furthermore, a parametric 3-dimensional finite element model, which allows modifications in the root length and its diameter, is described. For different geometries, the ideal moment-by-force ratios that induce a bodily movement were determined. The knowledge of root geometry is important in defining an optimal force system. PMID- 11941340 TI - Long-term stability of the leveling of the curve of Spee. AB - The aim of the study was to investigate whether the orthodontic leveling of the curve of Spee is a treatment procedure with a stable result on a long-term basis. Measurements were made on the plaster casts of 149 orthodontically treated patients (57 males and 92 females). The mean age before treatment was 12.8 years (range, 8-25 years). Study casts were taken before treatment (T1), at the completion of orthodontic therapy (T2), and 6.7 years (mean) posttreatment (T3). Inclusion criteria were no extractions, all Angle classifications except Class III malocclusions, and all permanent teeth fully erupted except second and third molars. The curve of Spee and the irregularity index were measured on standardized digital photographs of the casts. Overjet and overbite were assessed with a ruler. Changes in the curve of Spee were correlated with changes in irregularity index, overjet, and overbite from T1 to T3. The following results were noted: (1) leveling of the curve of Spee is a relatively stable treatment procedure compared with a return of incisor crowding and deepening of the bite; (2) neither the initial depth of the curve of Spee nor the initial irregularity index is an indicator for the amount of relapse; (3) the amount of leveling is not correlated with the relapse of the 4 tested parameters (curve of Spee, irregularity index, overjet, and overbite); and (4) there is a mild correlation between the relapse of the curve of Spee and the relapse of the irregularity index, overjet, and overbite. According to the results of the study, leveling the curve of Spee during orthodontic treatment seems to be very stable on a long-term basis; it was weakly correlated with the other variables tested. PMID- 11941341 TI - Therapeutic efficacy of an oral appliance in the treatment of obstructive sleep apnea: a 2-year follow-up. AB - The purpose of this study was to investigate the long-term efficacy of an oral appliance, the Karwetzky activator, on respiratory and sleep parameters in patients with obstructive sleep apnea (OSA). Those selected for this study were 26 patients polysomnographically diagnosed with mild-to-moderate OSA. They were initially treated successfully with this appliance, as documented by a second polysomnographic study after 6 to 12 weeks. Further polysomnographic registrations 6 to 12 months and 18 to 24 months later were performed for each patient wearing the appliance. For 21 patients (81%), therapeutic efficacy was maintained; 5 patients (19%) showed a deterioration in respiratory parameters. We corrected this by adjusting the device in 2 patients. The mean apnea-hypopnea index decreased significantly from 17.8 events per hour at the baseline registration to 4.2 events per hour (P <.001) after 6 to 12 weeks of treatment. After 6 to 12 months, the apnea-hypopnea index was 8.2 events per hour. The index remained at this level 18 to 24 months later, with 8.3 events per hour. Mean oxygen saturation was not improved with the activator, but the number of desaturations had decreased at the 6-to-12 week review. Again, the improvement declined with time, but the number of oxygen desaturations was still significantly decreased at 18 to 24 months (P <.01). Although the respiratory parameters remained statistically improved throughout the study (P <.01), sleep architecture did not change statistically. In most patients, therapeutic efficacy was maintained at the 2-year follow-up, although there was a tendency for effectiveness to fall over time. We concluded that the Karwetzky activator may be an effective treatment alternative for patients with mild-to-moderate OSA, but therapy requires diligent and regular polysomnographic follow-ups. Further long term studies are needed to assess the continued efficacy of this oral appliance in treating OSA. PMID- 11941342 TI - Transition of molar relationships in different skeletal growth patterns. AB - This study investigated maxillary and mandibular growth differences and their effect on the changes in molar relationships from early transitional dentition to adult permanent dentition. Various landmarks were identified and measurements made on the longitudinal cephalometric radiographs of 40 people. The subjects were classified into 3 groups based on the amount of sagittal skeletal growth difference between the jaws. In group A, the mandible grew more than did the maxilla; in Group B, growth was about the same; and in group C, the maxilla grew more than did the mandible. The results revealed that skeletal growth differences between the jaws significantly influenced the changes in molar relationship during the transitional dentition, not only by translating basal bones but also by altering the amount of physiologic mesial shift in the dentition. However, the skeletal growth difference during the permanent dentition did not influence the changes in molar relationship. The sagittal growth difference between the jaws was largely absorbed by a dentoalveolar compensation. Tooth movement showed different characteristic features depending on the amount of the skeletal growth difference: (1) If the mandible grew more than did the maxilla, the growth difference was mostly absorbed by mesial displacement of the maxillary first molars and counterclockwise rotation of the occlusal plane. Anterior occlusion was adjusted by mesial displacement and labial inclination of the maxillary incisors and lingual inclination of the mandibular incisors. (2) If the maxilla grew more than did the mandible, the growth difference was mainly absorbed by mesial displacement of the mandibular molars. Then the maxillary molars showed minimal mesial displacement. The occlusal plane also showed minimal rotational change. Anterior occlusion was adjusted by lingual tipping of the maxillary incisors and mesial displacement and labial tipping of the mandibular incisors. PMID- 11941343 TI - Remnant amount and cleanup for 3 adhesives after debracketing. AB - The cleanup of remnant bonding adhesive from the enamel surface after debonding is an important factor for clinicians. The purposes of this study were to compare the weight, the surface area, and the cleanup times of remnant adhesive for a composite resin, Transbond (TB); a resin-modified glass ionomer, Fuji ORTHO LC bonded to enamel both conditioned (FOC) and nonconditioned (FONC); and a fluoride releasing composite resin, Advance (ADV), bonded to nonetched enamel. In addition, 2 qualitative methods for scoring remnant adhesive were compared with the quantitative weight and area data. Forty extracted human incisors were weighed, bonded with brackets, debonded, weighed, and photographed. Area was measured from the photographs with a sonic digitizer. Mean adhesive remnant weights differed between groups (analysis of variance [ANOVA], P =.02): The remnants from ADV and FOC were equal and both significantly heavier than the remnants from FONC; the weights of the TB remnants were intermediate between the heavier ADV and FOC remnants and the lighter FONC remnants. Mean remnant areas differed between groups (ANOVA, P =.03): The remnants from ADV were significantly larger than the remnants from TB and FONC, which were equal; the areas of the FOC remnants were intermediate between the larger ADV remnants and the smaller remnants from TB and FONC. Mean cleanup times also differed between groups (ANOVA, P <.001): TB and FOC had equal times that were significantly longer than the times for ADV and FONC, which were equal. Adhesives bonded to acid-etched or conditioned enamel took about 1 and a half times longer to clean up than did those bonded to nonetched enamel. When bonded to conditioned enamel, the resin modified glass ionomer had mean remnant adhesive weights, areas, and cleanup times statistically equivalent to TB. ADV had the fastest mean cleanup time per amount of remnant (ANOVA, P <.002). The graphs of scores for 2 qualitative methods used for scoring remnant amount did not closely resemble the graphs based on weight and area. The graph of a newly introduced qualitative method better reflected the area data. Weighing may be a useful method for quantifying remnant adhesive. PMID- 11941344 TI - Effect of time on the shear bond strength of cyanoacrylate and composite orthodontic adhesives. AB - The purpose of this study was to compare the effects of time on the shear bond strength of a cyanoacrylate system and a composite adhesive system (1) within half an hour after bonding the bracket to the tooth, and (2) after 24 hours from the time of bonding when the adhesive had achieved most of its bond strength. Eighty freshly extracted molars were collected and stored in a solution of 0.1% (weight/volume) thymol. The teeth were cleansed, polished, and randomly separated into 4 groups: (I), cyanoacrylate adhesive debonded within one-half hour from initial bonding; (II), cyanoacrylate adhesive debonded after 24 hours immersion in deionized water at 37 degrees C; (III), composite adhesive debonded within one half hour from initial bonding; (IV), composite adhesive debonded after 24 hours immersion in deionized water at 37 degrees C. The results of the analysis of variance comparing the 4 experimental groups (F = 12.68) indicated significant differences between them (P =.0001). The composite adhesive at 24 hours had significantly greater shear bond strength than did the other 3 groups. In general, the shear bond strengths were greater in the 2 groups debonded after 24 hours. This was true for both the cyanoacrylate ((-)x 7.1 +/- 3.3 megaPascals [MPa]) and the composite ((-)x 10.4 +/- 2.8 MPa) adhesives. On the other hand, the shear bond strength was significantly lower in the 2 groups debonded within one-half hour of their initial bonding. The bond strength of the cyanoacrylate adhesive ((-)x 5.8 +/- 2.4 MPa) was not significantly different from that of the composite ((-)x 5.2 +/- 2.9 MPa) adhesive. Our findings indicated that the cyanoacrylate and the composite adhesives tested have clinically adequate shear bond strengths at half an hour and at 24 hours after initial bonding. The clinician needs to consider the properties of each adhesive: eg, the need to use a curing light and the ability to have more working time with the composite adhesive versus no light but only a 5-second working time before the cyanoacrylate adhesive starts to set. PMID- 11941345 TI - Comparison of fluoride release protocols for in-vitro testing of 3 orthodontic adhesives. AB - The objective of this study was to compare the fluoride release of 3 orthodontic adhesives using disks and bracketed teeth with different storage protocols. The adhesives used were a resin-modified glass ionomer (RMGI) (Fuji Ortho LC; GC America, Aslip, Ill), a polyacid-modified composite resin (PMCR)(Assure; Reliance Orthodontic Products, Itasca, Ill), and a composite control, Transbond XT (3M Unitek, Monrovia, Calif). Metal brackets were bonded to the buccal and lingual surfaces of 120 extracted human premolars. Five plastic containers holding 4 teeth (8 brackets) were used for each adhesive protocol. The samples were stored in containers holding 4 mL of deionized water at 37 degrees C for 28 and 84 days for the brackets and disks, respectively. The bracketed samples released larger initial amounts of fluoride compared with the disk samples during the first 5 to 6 days for both fluoride-releasing adhesives. The PMCR (Assure) released more fluoride (mg/cm(2)/day) than did the RMGI (Fuji Ortho LC) in all protocols with the exception of daily protocols when values diminished below the RMGI values near the 24th day and between the 56th and the 70th days for the bracketed and disk samples, respectively. Inconsistent values for fluoride release were noted in the bracket and disk samples when compared with daily versus cumulative water changes. Daily water changes revealed higher fluoride release levels (brackets), but this trend was not evident in the disk samples. Daily water changes may yield more clinically relevant data on fluoride release. PMID- 11941346 TI - Expression of secretory proteins in oral fluid after orthodontic tooth movement. AB - Orthodontic treatment alters the expression of secretory proteins at the local level in bone and the oral cavity, but its systemic effects are not well understood. Total secretory proteins and a cyclic adenosine monophosphate (AMP) dependent protein kinase subunit (RII) were measured in saliva and gingival crevicular fluid (GCF) after the placement of orthodontic separators to determine if mechanical force applied to teeth affects protein secretion. Whole saliva and GCF were collected before and 1 day after treatment. Electrophoresis and Western blotting were carried out to establish the banding patterns of total proteins and to measure the isotype and amount of RII that serves as an apparent stress indicator. Digitized image files were used for densitometric analyses of the relative concentrations of RII and total protein. Individual protein values showed no statistically significant changes in saliva or GCF. Western blots, however, showed a dramatic difference in RII after the placement of separators: the 50-to-55 kilodaltons (kd) band virtually disappeared and was replaced by a fragment in the 20-kd range. These results suggest that although the expression of total proteins is not altered by mechanical force applied to teeth, a systemic response via the cyclic AMP signaling pathway might have been activated. PMID- 11941347 TI - Transposed and impacted maxillary canine with ipsilateral congenitally missing lateral incisor. PMID- 11941348 TI - A duck is a duck. PMID- 11941349 TI - Stafne's bone cavity and its utilization in complete denture retention. AB - This clinical report describes the fabrication of a mandibular denture for an elderly woman who presented with bilateral Stafne's bone cavities. A review of this anatomical entity is presented together with the prosthodontic management of the patient. PMID- 11941350 TI - Influence of different convergence angles and tooth preparation heights on the internal adaptation of Cerec crowns. AB - STATEMENT OF PROBLEM: Because of an imagining principle called active triangulation in the Cerec system, a shadow is cast distal to the illuminated objects. This distal shadow may be enlarged when the occlusal-cervical height of the prepared tooth is increased. Depth data of the shadow are unreliable, so the internal fit of Cerec crowns has been questioned. PURPOSE: This study evaluated the influence of different convergence angles and tooth preparation heights on the internal adaptation of Cerec crowns. MATERIAL AND METHODS: Tooth preparations were made on typodont teeth with different combinations of convergence angles and occlusal-cervical heights: Group I = 20 degrees angle, 6 mm height; Group II = 20 degrees angle, 4 mm height; Group III = 12 degrees angle, 6 mm height; and Group IV = 12 degrees angle, 4 mm height. Ten Cerec crowns were fabricated for each type of tooth preparation. Measurements of the internal fit were performed with the cement space replica technique and an image analysis system. Three-way analysis of variance was used to analyze the differences in cement space with different tooth preparations and the number of times that milling tools were used to prepare the Cerec crowns (P<.05). Multiple comparisons were made to evaluate differences between groups (P<.0083). RESULTS: Cerec crowns with a 12 degrees convergence angle demonstrated the best internal fit (cement space in Groups III and IV = 121 +/- 41 microm and 115 +/- 42 microm, respectively). The difference between the 2 convergence types was within the range of the scanning error (25 microm) produced by the Cerec camera. The number of times that milling tools were used had no significant effect on internal fit (P=.78). Tooth preparation height equal to or shorter than 6 mm occlusal-cervically with both 12 degrees and 20 degrees convergence angles also had no significant effect on internal fit (P>.0083). Cement space at distal walls (185 +/- 28 microm) was the thickest among all axial walls (P=.0001) and was twice as thick as that at the facial (90 +/- 14 microm) and palatal walls (92 +/- 15 microm). CONCLUSION: Within the limitations of this study, there was little difference in the internal fit of Cerec crowns prepared with convergence angles of 12 degrees and 20 degrees. Distal shadows influenced the thickness of the cement spaces, particularly at the distal walls. However, tooth preparations with an occlusal-cervical height not greater than 6 mm did not exaggerate the effect of the distal shadows. PMID- 11941351 TI - Relationship between crown placement and the survival of endodontically treated teeth. AB - STATEMENT OF PROBLEM: Crowns have been considered the restoration of choice for endodontically treated teeth, but their selection has been based primarily on anecdotal evidence. PURPOSE: This study tested the hypothesis that crown placement (coronal coverage) is associated with improved survival of endodontically treated teeth when preaccess, endodontic, and restorative factors are controlled. MATERIAL AND METHODS: A University of Iowa College of Dentistry treatment database was used to identify permanent teeth that had undergone initial obturation between July 1, 1985, and December 31, 1987. Study patients were restricted to persons with at least 1 dental visit in each 2-year interval from 1985 to 1996; a simple random sample of 280 patients (n = 400 teeth) was selected. Dental charts, radiographs, and computerized databases were examined to ascertain variables of interest and to verify study inclusion criteria. Kaplan Meier survival estimates were generated for the 203 teeth that satisfied study inclusion criteria. Multivariate Cox proportional hazards regression models were developed, with standard errors adjusted to account for clustering of teeth within patients. RESULTS: When tooth type and radiographic evidence of caries at access were controlled, the final Cox model showed that endodontically treated teeth not crowned after obturation were lost at a 6.0 times greater rate than teeth crowned after obturation (95% confidence interval: 3.2 to 11.3). CONCLUSION: Within the limitations of this study, a strong association between crown placement and the survival of endodontically treated teeth was observed. These results may impact treatment planning if long-term tooth retention is the primary goal. PMID- 11941352 TI - The effect of water storage and light exposure on the color and translucency of a hybrid and a microfilled composite. AB - STATEMENT OF PROBLEM: Internal discoloration may occur with the intraoral use of resin-based restoration materials. Water storage and light exposure influence the color properties of microfilled and hybrid composites. PURPOSE: The purpose of this study was to determine color and translucency changes in a hybrid and a microfilled composite after light exposure with and without water storage. MATERIAL AND METHODS: A hybrid (Tetric) and a microfilled composite (Silux Plus) were subjected to artificial daylight with and without water storage. Tristimulus Yxy values were determined colorimetrically against a black or a white background. Differences from baseline were calculated as Delta E*(ab) for up to 1 month. After 1 month, Delta L*, Delta a*, Delta b*, and the contrast ratio Delta C were calculated. Data were analyzed with 2-way analysis of variance (P<.05). RESULTS: The Delta E(ab)* increased over time for wet and dry stored specimens of both materials. Wet storage for 1 month resulted in significantly higher Delta E(ab)* and Delta C but lower Delta b* than dry storage. The Delta a* was significantly higher for Tetric compared with Silux Plus. Both materials showed negative Delta L* values under both storage conditions without significant differences between materials or storage conditions. CONCLUSION: The results of this in vitro study suggest that resin-based restoration materials undergo measurable changes due to daylight exposure. Increased changes occurred under the influence of water storage. PMID- 11941353 TI - Tensile bond strengths of composites to a gold-palladium alloy after thermal cycling. AB - STATEMENT OF PROBLEM: Many different materials and methods have been used to fabricate or repair veneer facings with composites, but only a few of these have been studied. PURPOSE: This study compared the tensile bond strengths of composites to a gold-palladium alloy with the use of several surface treatment methods. MATERIAL AND METHODS: Forty alloy specimens were cast in Eclipse (52% gold and 37.5% palladium) in the form of truncated cones. These specimens were divided equally into 4 groups. In group I, the bonding surfaces of the metal cones were treated with Silicoater MD. Truncated cones of Dentacolor composite were bonded to the metal surfaces and light-polymerized. In group II, the bonding surfaces of the metal cones were air-particle abraded with 50 microm aluminum oxide and coated with C&B Metabond. Truncated cones of Epic-TMPT composite were bonded to the metal surfaces and light-polymerized. In group III, the bonding surfaces of the metal cones were air-particle abraded with CoJet-Sand. Truncated cones of Pertac-Hybrid composite were bonded to the metal surfaces and light polymerized. In group IV, the bonding surfaces of the metal cones were air particle abraded with CoJet-Sand. Truncated cones of Visio-Gem were bonded to the metal surfaces and light-polymerized. After 24 hours of water immersion at 37 degrees C and 1000 thermal cycles in water at 5 degrees C and 55 degrees C, tensile forces were applied to all specimens with a universal testing machine. Analysis of variance was applied to the data (P<.05), and differences among means were determined with a Tukey-Kramer interval of 5.4 MPa. RESULTS: Tensile bond strengths in MPa were as follows: Dentacolor, 14 +/- 5; Epic-TMPT, 12 +/- 4; Pertac-Hybrid, 13 +/- 5; and Visio-Gem, 18 +/- 4. The tensile bond strength of Visio-Gem was significantly higher than that of Epic-TMPT, but no differences were found among Dentacolor, Pertac-Hybrid, and Epic-TMPT (P<.05). CONCLUSION: Within the limitations of this study, all 4 bonding systems tested produced high bond strengths between composites and a gold-palladium alloy after thermal cycling. PMID- 11941354 TI - Casting titanium partial denture frameworks: a radiographic evaluation. AB - STATEMENT OF PROBLEM: Titanium is the most biocompatible metal available for dental castings. However, there is some concern about the castability of titanium used on a daily basis. PURPOSE: A radiographic evaluation of titanium partial denture frameworks was undertaken to ascertain whether these castings were technically acceptable for clinical use. MATERIAL AND METHODS: Three hundred Grade II titanium removable partial denture frameworks were cast in a Titec 205M casting machine. All materials were used as directed by the manufacturer, and all castings were made by an experienced titanium specialist. Each casting was evaluated by radiograph. Based on the number, location, and size of argon inclusions, the frameworks were rated as follows: technically acceptable for clinical use as cast; technically acceptable after laser welding modifications; or unacceptable as cast, with remake necessary. RESULTS: Of the 300 titanium frameworks cast, 97% were rated technically acceptable for clinical use in terms of castability. SUMMARY: Within the limitations of this radiographic evaluation, it was determined that the castability of titanium was such that technically acceptable castings could be made on a daily basis. PMID- 11941355 TI - A review of guidelines for implant rehabilitation of the edentulous maxilla. AB - This review suggests some guidelines and protocols for treatment of the edentulous maxilla with osseointegrated implants. Evidence-based treatment options are discussed with reference to risk assessment. Developing technologies applicable to diagnosis, clinical procedures, and laboratory techniques are also discussed. Osseointegrated implants are increasingly advocated as a treatment option for maxillary edentulism and in many situations are the treatment of choice. PMID- 11941356 TI - Multifactorial modeling of temporomandibular anatomic and orthopedic relationships in normal versus undifferentiated disk displacement joints. AB - STATEMENT OF PROBLEM: There is persistent dispute about the diagnostic value of hard tissue anatomic relationships in predicting temporomandibular joint disorders and normals. PURPOSE: The goal of this study was identification of multifactorial temporomandibular hard tissue relationships that differentiate asymptomatic normal joints. MATERIAL AND METHODS: Central section lateral tomograms of 162 female temporomandibular joints with pooled diagnoses of unilateral disk displacement with and without reduction were compared to 42 female asymptomatic normal joints using 14 linear and angular measurements and 8 ratios. A validated classification tree model was tested for accuracy with sensitivity, specificity, goodness of fit, and the amount of the log likelihood accounted for. The tree model was compared with a multiple logistic regression model and univariate testing. RESULTS: The classification tree model consisted of 3 asymptomatic and 4 disk displacement terminal nodes consisting of interactions of condyle position with measures of fossa size and shape, of which mainly average non-extreme measurements and more frequent concentric ranges typified the asymptomatic joints. The logistic regression and univariate models also incorporated condyle position and size, but the logistic regression accounted for less of the log likelihood than the tree (23.3% vs. 32.6% Rescaled Cox and Snell R(2)). The tree and the logistic regression models were moderately good predictors for distinguishing normals from disk displacement joints (sensitivity 67.9% and 72.2%, specificity 85.7% and 76.2%, respectively). Although the univariate analysis showed that the asymptomatic joints had smaller mean fossa width to fossa depth ratios (P<.0005), shorter mean eminence length (P<.007), and more concentric to anterior mean condyle position (P<.049), overlap in most of the ranges limited the predictive value. CONCLUSION: Within the limitations of this study, multifactorial analysis revealed that several subsets of asymptomatic temporomandibular joints could be distinguished from joints with disk displacement according to hard tissue measurements taken from central section tomograms. In general, asymptomatic normal joints were typified by interactions of less extreme ranges of fossa size, shape, and condyle position. PMID- 11941357 TI - Multifactorial comparison of disk displacement with and without reduction to normals according to temporomandibular joint hard tissue anatomic relationships. AB - STATEMENT OF PROBLEM: There is disagreement about the predictive value of temporomandibular joint tomographic anatomy in the diagnosis of internal derangements. PURPOSE: This study aimed to identify multifactorial temporomandibular hard tissue relationships that differentiate disk displacement with reduction and disk displacement without reduction from normals. MATERIAL AND METHODS: Temporomandibular joint tomograms from females diagnosed with unilateral disk displacement with (n=84) or without (n=78) reduction were compared to 42 asymptomatic normal joints with the use of 14 linear and angular measurements and 8 ratios. A validated classification tree model was tested for accuracy with sensitivity, specificity, goodness of fit, and the amount of log likelihood accounted for. The tree model was compared with a multiple logistic regression model and univariate testing. RESULTS: The disk displacement with reduction tree model consisted of 3 disease and 2 normal pathways with interactions between fossa width to depth ratio, condyle position, and linear posterior joint space. This class was characterized by either a much wider- and shallower-than-average fossa shape and/or by a moderately posterior condyle position when the fossa shape was average to deeper and/or narrower. The logistic regression and univariate models also suggested wider and/or shallower fossae, as well as longer eminence length. The disk displacement without reduction tree model consisted of 2 disease pathways and 1 normal pathway. Interactions characterized this class by either a posterior to very posterior condyle position or by a much deeper than average fossa depth when the condyle position was concentric to anterior. The logistic regression model emphasized greater fossa depth and width versus normals. The tree models conservatively predicted the disease classes: Rescaled Cox and Snell R(2) 37.0%, sensitivity 70.2%, and specificity 90.5% for disk displacement with reduction; R(2) 28.8%, sensitivity 66.7%, and specificity 85.7% for disk displacement without reduction. CONCLUSION: Within the limitations of this study, hard tissue relationships revealed by central tomogram sections were able to model notable differences between disk displacement with reduction and disk displacement without reduction versus asymptomatic normals when temporomandibular joints were examined as a multifactorial system typified by interactions of fossa width to depth proportions and condyle position. While substantial, the hard tissue predicted only part of the biology. The model could be broadened by additional factors and interactions. PMID- 11941358 TI - Effect of finishing and polishing procedures on the gap width between a denture base resin and two long-term, resilient denture liners. AB - STATEMENT OF PROBLEM: The junction between a long-term, resilient denture liner and the denture base is difficult to finish and polish due to differences in the hardness of the materials. Gaps tend to form during finishing and polishing procedures. PURPOSE: This study measured the junctional gap between 2 long-term, resilient denture liners and a denture base material after different finishing and polishing procedures were performed. The surface smoothness of the 2 liner materials also was evaluated. MATERIAL AND METHODS: Molloplast-B and an experimental, heat-polymerized methyl siloxane-resin-based denture liner were processed (according to the manufacturer's instructions) against Lucitone 199 acrylic resin. Control specimens (n = 20) were 2 mm thick and flat. Experimental specimens (n = 64) were fabricated with a raised center section: a 3- x 5- x 15 mm half-cylindrical ridge with a junction at the top of the cylinder between the liner material and acrylic resin. The specimens were finished with 1 of 6 types of burs and polished with different combinations of rubber-impregnated acrylic polishers, pumice, and tin oxide. The finished specimens were examined and photographed with a scanning electron microscope, and the largest gap along the liner/denture base junction on each specimen was measured. Three-way analysis of variance without replication (P<.05) and post-hoc t tests were used to analyze the data and compare groups. RESULTS; Averaged across finishing and polishing techniques, a larger gap was recorded for the experimental liner material (22 microm) than for Molloplast-B (14 microm) (P<.00005). Qualitative evaluation suggested that the experimental liner material polished better than Molloplast-B. The smoothest surfaces were obtained when specimens were finished with fine-tooth cross-cut carbide burs and polished with both pumice and tin oxide. A comparison between polished and unpolished specimens, averaged over materials and finishing techniques, revealed that polishing reduced gap size (P=.015). CONCLUSION: Within the limitations of this study, the 2 denture liners tested behaved similarly. The results suggest that the size of the gap at the liner/denture base junction may be affected by finishing and polishing techniques and vary among materials. PMID- 11941360 TI - Evaluation of a device for attenuation of electron release from dental restorations in a therapeutic radiation field. AB - STATEMENT OF PROBLEM: For some patients, radiation treatment is a part of tumor therapy in the head and neck area before and/or after surgery. The oral cavity and teeth are thereby frequently exposed to high doses of radiation. In this situation, electronic backscatter from dental materials may damage the surrounding soft tissue. PURPOSE: This study determined the degree of absorption and the backscatter effect of therapeutic radiation used in the presence of 4 different dental materials. The efficacy of a protective stent also was investigated. MATERIAL AND METHODS: The influence of 4 dental materials (a high gold alloy, pure titanium, amalgam, and a synthetic material) on radiation dose distribution was tested on 2 test models that simulated the presence of teeth. An alanine dosimeter was used to make measurements with and without the presence of a protective stent. To verify the results, one of the test models was compared to a computer simulation. RESULTS: Backscatter effects on the surface of dental materials caused an increase of up to 170% of the radiation dose measured without the materials. The rate of overdose increased with the atomic number of the dental material. The extent of the backscatter effect was a maximum of 4 mm. CONCLUSION: The considerable overdose of 170% found in this study suggests that soft tissue surrounding dental restorations should be protected from radiation. The backscatter results indicate that soft tissue could be effectively shielded with a 3-mm synthetic stent. PMID- 11941359 TI - Abrasion resistance of a resin-impregnated type IV gypsum in comparison to conventional products. AB - STATEMENT OF PROBLEM: Limited abrasion resistance is one of the disadvantages of gypsum materials. One means to improve these properties may be the addition of various polymers to reinforce surface and subsurface wear resistance. PURPOSE: The purpose of this study was to evaluate the abrasion resistance of 3 types of gypsum materials with a novel abrasion wear device. MATERIAL AND METHODS: A standard test specimen (50 x 50 x 8 mm) with repeating 1 mm, 45 degrees -angle vertical ridges was used as the test surface. Three types of gypsum materials were evaluated: a type III stone, a conventional type IV stone, and a resin impregnated type IV stone. Specimens of each material (n = 30) were poured in a vinyl polysiloxane impression of the standard test specimen with the use of an acrylic resin custom tray. Each gypsum product was mixed according to the manufacturer's instructions, poured, and separated 1 hour later. Specimens were trimmed and allowed to bench set for 24 hours (n = 15) or 7 days (n = 15). Abrasion was evaluated with the use of a reciprocal moving stage positioned below a weighted stylus. The stylus was loaded with 3 different weights (15, 50, and 75 g), and the individual gypsum specimens were moved reciprocally for 10 passes. Before and after each set of passes, the specimen was measured on an analytical balance to determine material loss. Three-way ANOVA and Scheffe's multiple comparisons test were used to analyze the data (P<.05). RESULTS: Type III gypsum at 24 hours exhibited significantly greater material loss than type IV stones at both 7 days and 24 hours (P<.001). Type III gypsum specimens exhibited significantly greater material loss at 24 hours than at 7 days (P<.001). There was no significant difference in the abrasion resistance of type IV specimens at 24 hours and 7 days. CONCLUSION: Within the limitations of this study, no significant differences in abrasion resistance were found between specimens fabricated with resin-impregnated and conventional type IV stones, although the former exhibited the least material loss. PMID- 11941361 TI - A procedure for making trial dentures. AB - A procedure is presented for making trial dentures by arranging denture teeth in soft wax rather than hard baseplate wax. The drawbacks of using hard wax are thereby avoided. PMID- 11941362 TI - Surgical oral radiographic guide with a removable component for implant placement. AB - Radiographic guides are essential for optimal implant positioning. Surgical guides permit implants to be placed in preselected positions and at proper angulations. This article describes a procedure for combining the functions of radiographic and surgical guides in a single device. The radiographic component is easily separated from the surgical component after the necessary radiographs have been made. This procedure results in less paraphernalia and reduces costs since only 1 basic guide with an add-on component is fabricated. PMID- 11941364 TI - Locking retentive attachment for an implant-retained auricular prosthesis. AB - This article describes the use of the Slant-Lock retention system with an implant retained auricular prosthesis. PMID- 11941363 TI - The use of healing abutments for the fabrication of cement-retained, implant supported provisional prostheses. AB - This article describes the intraoral preparation of healing abutments for use as prefabricated abutments for a cement-retained, implant-supported prosthesis. After the healing abutments are prepared, an impression is made with irreversible hydrocolloid, and the provisional restoration is fabricated indirectly. This technique is an easy and economical alternative for the fabrication of provisional fixed partial dentures or crowns but may be contraindicated for severely misaligned implants. PMID- 11941365 TI - Fabrication of a provisional restoration after fracture of an anterior crown. PMID- 11941366 TI - An indirect technique for fabrication of a post and core wax pattern. PMID- 11941367 TI - A modified implant impression technique. PMID- 11941368 TI - A reverse-angle dental mirror. PMID- 11941369 TI - Mutations in ALMS1 cause obesity, type 2 diabetes and neurosensory degeneration in Alstrom syndrome. AB - Alstrom syndrome is a homogeneous autosomal recessive disorder that is characterized by childhood obesity associated with hyperinsulinemia, chronic hyperglycemia and neurosensory deficits. The gene involved in Alstrom syndrome probably interacts with genetic modifiers, as subsets of affected individuals present with additional features such as dilated cardiomyopathy, hepatic dysfunction, hypothyroidism, male hypogonadism, short stature and mild to moderate developmental delay, and with secondary complications normally associated with type 2 diabetes, such as hyperlipidemia and atherosclerosis. Our detection of an uncharacterized transcript, KIAA0328, led us to identify the gene ALMS1, which contains sequence variations, including four frameshift mutations and two nonsense mutations, that segregate with Alstrom syndrome in six unrelated families. ALMS1 is ubiquitously expressed at low levels and does not share significant sequence homology with other genes reported so far. The identification of ALMS1 provides an entry point into a new pathway leading toward the understanding of both Alstrom syndrome and the common diseases that characterize it. PMID- 11941370 TI - Mutation of ALMS1, a large gene with a tandem repeat encoding 47 amino acids, causes Alstrom syndrome. AB - Alstrom syndrome (OMIM 203800) is an autosomal recessive disease, characterized by cone-rod retinal dystrophy, cardiomyopathy and type 2 diabetes mellitus, that has been mapped to chromosome 2p13 (refs 1-5). We have studied an individual with Alstrom syndrome carrying a familial balanced reciprocal chromosome translocation (46, XY,t(2;11)(p13;q21)mat) involving the previously implicated critical region. We postulated that this individual was a compound heterozygote, carrying one copy of a gene disrupted by the translocation and the other copy disrupted by an intragenic mutation. We mapped the 2p13 breakpoint on the maternal allele to a genomic fragment of 1.7 kb which contains exon 4 and the start of exon 5 of a newly discovered gene (ALMS1); we detected a frameshift mutation in the paternal copy of the gene. The 12.9-kb transcript of ALMS1 encodes a protein of 4,169 amino acids whose function is unknown. The protein contains a large tandem-repeat domain comprising 34 imperfect repetitions of 47 amino acids. We have detected six different mutations (two nonsense and four frameshift mutations causing premature stop codons) in seven families, confirming that ALMS1 is the gene underlying Alstrom syndrome. We believe that ALMS1 is the first human disease gene characterized by autosomal recessive inheritance to be identified as a result of a balanced reciprocal translocation. PMID- 11941371 TI - The TRPM7 channel is inactivated by PIP(2) hydrolysis. AB - TRPM7 (ChaK1, TRP-PLIK, LTRPC7) is a ubiquitous, calcium-permeant ion channel that is unique in being both an ion channel and a serine/threonine kinase. The kinase domain of TRPM7 directly associates with the C2 domain of phospholipase C (PLC). Here, we show that in native cardiac cells and heterologous expression systems, G alpha q-linked receptors or tyrosine kinase receptors that activate PLC potently inhibit channel activity. Numerous experimental approaches demonstrated that phosphatidylinositol 4,5-bisphosphate (PIP(2)), the substrate of PLC, is a key regulator of TRPM7. We conclude that receptor-mediated activation of PLC results in the hydrolysis of localized PIP(2), leading to inactivation of the TRPM7 channel. PMID- 11941372 TI - Frodo interacts with Dishevelled to transduce Wnt signals. AB - Dishevelled (Dsh) is required for the specification of cell fate and polarity by secreted Wnt proteins. Frodo, a novel conserved Dsh-binding protein, synergized with Xenopus Dsh (XDsh) in secondary axis induction in Xenopus laevis embryos. A dominant inhibitory construct and antisense oligonucleotide-mediated depletion of Frodo inhibited axial development in response to XDsh and XWnt8, and suppressed transcriptional activation of a reporter construct. At later embryonic stages, both dominant negative Frodo and antisense oligonucleotides interfered with the expression of regional neural markers and caused eye deficiencies, indicating that Frodo is required for normal eye and neural tissue development. Full-length Frodo RNA suppressed these loss-of-function phenotypes, attesting to their specificity. These findings establish a function for Frodo as an essential positive regulator of Wnt signalling. PMID- 11941373 TI - Perceiving patterns in random series: dynamic processing of sequence in prefrontal cortex. AB - We demonstrate that regions within human prefrontal cortex develop moment-to moment models for patterns of events occurring in the sensory environment. Subjects viewed a random binary sequence of images, each presented singly and each requiring a different button press response. Patterns occurred by chance within the presented series of images. Using functional magnetic resonance imaging (fMRI), we identified activity evoked by viewing a stimulus that interrupted a pattern. Prefrontal activation was evoked by violations of both repeating and alternating patterns, and the amplitude of this activation increased with increasing pattern length. Violations of repeating patterns, but not of alternating patterns, activated the basal ganglia. PMID- 11941374 TI - Immunization reverses memory deficits without reducing brain Abeta burden in Alzheimer's disease model. AB - We have previously shown that chronic treatment with the monoclonal antibody m266, which is specific for amyloid beta-peptide (Abeta), increases plasma concentrations of Abeta and reduces Abeta burden in the PDAPP transgenic mouse model of Alzheimer's disease (AD). We now report that administration of m266 to PDAPP mice can rapidly reverse memory deficits in both an object recognition task and a holeboard learning and memory task, but without altering brain Abeta burden. We also found that an Abeta/antibody complex was present in both the plasma and the cerebrospinal fluid of m266-treated mice. Our data indicate that passive immunization with this anti-Abeta monoclonal antibody can very rapidly reverse memory impairment in certain learning and memory tasks in the PDAPP mouse model of AD, owing perhaps to enhanced peripheral clearance and (or) sequestration of a soluble brain Abeta species. PMID- 11941375 TI - Analgesia and public health: meeting the global challenges. PMID- 11941376 TI - Meeting consumer demands for analgesics. AB - Responsible self-medication provides a positive health outcome for the individual and benefits for society as a whole. Nonprescription analgesics are among the most widely used drugs for self-medication, playing a key role in the management of a number of common disorders. While ensuring that the benefits of self medication are available to consumers and to society, it is important to acknowledge the potential risks inherent in the widespread and unsupervised use of medicines. Along with drugs that are well tolerated and effective in normal use, responsible self-medication depends on providing consumers with the information they need to use drugs appropriately in a proven usable form, and it can benefit from the advice and guidance offered by health care professionals. PMID- 11941377 TI - Evaluating analgesia: the challenges. AB - Decisions must be made when assigning analgesics, and such decisions involve comparing efficacy and adverse effects. The rules for assessing efficacy by systematic review and by large clinical trials are becoming clearer all the time. It is known, for instance, that trials that are not randomized, are not double blind, or are too small will exaggerate efficacy. These rules are relatively easy to apply to drug interventions, although there are glaring exceptions, such as in early intra-articular morphine studies. The rules may also be less easy to apply for analgesics available over-the-counter (OTC) if the drugs have not been studied in trials of high quality or if the OTC-recommended doses are below the doses studied in the trials. When there are at least 500 patients studied in trials of high quality, credible efficacy estimates for effective drugs can be derived; more patients are needed for less effective drugs. The number needed to treat and the number needed to harm can be used to compare the efficacy and safety of a treatment with placebo or with other treatments. A recent meta analysis comparing the number needed to treat for paracetamol 1000 mg and paracetamol 600-650 mg in moderate to severe postoperative pain indicated that paracetamol 1000 mg is likely to be more effective than lower doses. Large data sets allow investigation into dose response, gender differences, and whether particular drugs work better for particular painful conditions. Purists would argue that databases of patient experience should not be used for these efficacy analyses. The rules on how to collect and use evidence on adverse effects are less clearly formulated than they are for efficacy. For safety at or above therapeutic dose, the rules of evidence-gathering are necessarily different, and randomized trials are rarely an adequate or sufficient source. Databases of patient experience, with all the caveats, may be the most reliable data from which to work. PMID- 11941378 TI - Analgesia and the patient with osteoarthritis. AB - Osteoarthritis is the most common form of arthritis, its prevalence increasing with age: as much as 80% of the population over 75 years show radiologic signs of the condition. Symptoms include pain, stiffness, and functional impairment; however, not all patients are symptomatic. Management starts with nonpharmacologic interventions, followed by pharmacologic means, and ultimately by surgical intervention. The management is multidisciplinary and is tailored to the needs of the individual patient. It is, therefore, a good model of collaborative care: multidisciplinary management of a chronic condition for which the patients themselves coordinate the use of the management options, with information and guidance from health care professionals and written materials, as needed. Guidelines for the management of osteoarthritis have been developed and are applied in different continents. These guidelines are based on searches of the literature and evidence-based interpretation, in combination with expert opinion. Pharmacologic management guidelines state that based on its overall efficacy, toxicity profile, and cost, paracetamol-acetaminophen should be tried first and, if successful, should be used as the preferred long-term analgesic. In patients who do not experience adequate symptomatic relief with paracetamol acetaminophen, alternative or additional pharmacologic agents should be considered, especially nonsteroidal anti-inflammatory drugs (NSAIDs). When the combination of paracetamol-acetaminophen with NSAIDs fails, tramadol may be given. Most patients with osteoarthritis are able, after discussion with their physician, to manage their symptoms themselves. They make use of educational occupational, and physical advisers, and they use their medication on demand. The basis of this self-administered pharmacologic management is paracetamol acetaminophen, sometimes in combination with NSAIDs. A promising option for the future is the development of symptomatic slow-acting drugs for osteoarthritis that possess structure-modifying properties. PMID- 11941379 TI - Confounding by indication in epidemiologic studies of commonly used analgesics. AB - Confounding by indication is a bias frequently encountered in observational epidemiologic studies of drug effects. Because the allocation of treatment in observational studies is not randomized and the indication for treatment may be related to the risk of future health outcomes, the resulting imbalance in the underlying risk profile between treated and comparison groups can generate biased results. Confounding by indication is often present in studies of drugs that are not widely prescribed, because the indications for their use are narrow and not likely to be present in comparison groups; however, this bias is also observed in the study of widely used over-the-counter and prescription drugs, are exemplified by studies of analgesics. In this article we review examples from the published literature to demonstrate how confounding by indication can affect the findings of pharmacoepidemiologic studies relating analgesic use to various health outcomes. PMID- 11941380 TI - Analgesia and public health: what are the challenges? AB - Pain is a common problem, but unfortunately, it is one that is still notoriously neglected and poorly managed. Although it usually is not rated highly in public health statistics, it forms a substantial proportion of the everyday work of health care professionals, and thus remains a major public health burden. The first challenge in successful pain management is overcoming the ineffective learning processes most health care practitioners use to update their procedures and therapies in response to the latest research. The ready availability of over the-counter analgesics means that much of the pain in the community is now self medicated, and it is vital that they also have ready access to the latest evidence-based recommendations. Second, better methods are needed to tailor treatment to individual patients because differences in comorbidities, drug metabolism, or the nature and severity of disease processes lead to different responses from individual patients. Such tailoring should also account for differences in side-effect profiles of the various treatment options available. Finally, even if health practitioners are aware of the latest in clinical evidence and recommended practices, they may not be able to implement the most appropriate treatment because of legal or financial barriers. This article will to review these three challenges to the management of pain and discuss practical ways in which they may be handled to help reduce the burden of pain care in society. PMID- 11941381 TI - Next generation of everyday analgesics. AB - Pain is the leading symptom of most diseases. Humans have always tried to overcome pain using physical and chemical means, and it is believed that opioids and salicylates present in natural products have been used since prehistoric times. The development of the sciences, in particular chemistry and medicine, in the 19th century led to the discovery of the active ingredients of poppy and willow bark (morphine and salicylic acid). Shortly after, synthetic chemistry provided substitutes produced from coal tar (ie, acetaminophen, aspirin, phenazone, and pethidine). These represent the two main types of analgesics commonly used to treat mild and serious pain: the opioids (pethidine) and the antipyretic analgesics, which may be further divided into the aspirin-derived (acidic) nonsteroidal anti-inflammatory drugs (eg, ibuprofen) and the phenazone and acetaminophen-like (nonacidic) antipyretic analgesics (which have little anti inflammatory activity). Chemical modifications and broad-spectrum screening provided medicine with thousands of pharmacologic analogs that broadened the therapeutic spectrum but did not supplant the original compounds developed in the 19th and early 20th century. Recently, molecular biology and genomics have led to the development of new target-selective chemical entities for use in pain relief. These include selective cyclooxygenase (COX)-2 inhibitors, substance P, blockers or agonists of cannabinoid and vanilloid receptors, inhibitors of tetradotoxin resistant Na channels, and many more. Most of these selective compounds did not succeed in everyday pain treatment. Some look promising, including the COX-2 selective inhibitors, but doubts remain about the superiority of these new compounds in everyday use. This is particularly the case with the generation of selective COX-2 inhibitors currently in clinical use. PMID- 11941382 TI - Analgesics and glutathione. AB - A growing body of evidence indicates that glutathione (GSH) plays a vitally important role in cellular function. It detoxifies toxic metabolites of drugs and reactive oxygen species and regulates gene expression, apoptosis, and transmembrane transport of organic solutes. The maintenance of GSH homeostasis is essential for the organism to perform its many functions. The turnover of GSH is a dynamic process, and large quantities of GSH are synthesized per day from its precursor amino acids cysteine, glutamic acid, and glycine. Toxic doses of paracetamol deplete intracellular GSH and result in cell death by a combination of mechanisms, leading to necrosis and apoptosis, mainly in the liver. In clinical situations characterized by low GSH, the risk of toxicity from therapeutic doses of paracetamol may conceivably be increased. This toxicity has been reported in chronic alcoholics who have low intrahepatic GSH and who may have an induced enzyme system that generates the toxic metabolite of paracetamol. Considering the large number of alcoholics in our population and the widespread use of paracetamol, this must be a rare and essentially unpredictable occurrence. Except for anecdotal reports, there is no convincing evidence that other populations in which low GSH has been observed-such as patients with human immunodeficiency virus (HIV) infection or chronic hepatitis C, malnourished patients, and patients with cirrhosis-are at higher risk of experiencing adverse events from paracetamol. PMID- 11941383 TI - Analgesics and asthma. AB - The incidence of asthma is increasing throughout the world, which presents both public health and economic concerns. It is widely recognized that in some adult patients with asthma, aspirin and other nonsteroidal anti-inflammatory drugs (NSAIDs) that inhibit cyclooxygenase (COX)-1 exacerbate the condition. This is a distinct clinical syndrome called aspirin-induced asthma (AIA). The disease develops according to a characteristic pattern of symptoms. Persistent eosinophilic rhinosinusitis precedes development of nasal polyposis, aspirin hypersensitivity, and asthma. There is no in vitro test, and diagnosis can only be established by provocation tests with aspirin. At the biochemical level, AIA is characterized by a chronic overproduction of cysteinyl leukotrienes. The key enzyme, leukotriene C4 synthase, is overexpressed in bronchi, and its messenger RNA is upregulated in peripheral blood eosinophils. This can be partly related to the genetic polymorphism of the enzyme. The disease runs a protracted course, even if COX-1 inhibitors are avoided. The course of AIA is often severe, and at least half of the patients need systemic corticosteroids to control their asthma. To prevent life-threatening reactions, patients with AIA should avoid aspirin and other analgesics that inhibit COX-1. The incidence of cross-sensitivity to paracetamol in AIA patients is low and, when a reaction does occur, the symptoms experienced are shorter and milder than if the reactions were evoked by an NSAID. Rapidly growing evidence indicates that highly specific COX-2 inhibitors, known as coxibs, are well tolerated and can be safely used by AIA patients. PMID- 11941384 TI - Over-the-counter analgesics: a toxicology perspective. AB - The decision to use any analgesic is a balance of benefit and risk. In the case of analgesics, it is important to balance the therapeutic benefit against both the risk in therapeutic use and the risk (and ease of treatment) in overdose. Paracetamol in therapeutic dose carries little risk of adverse events. Less than 0.1% of the estimated 30 million paracetamol users in the United Kingdom attend hospital with a paracetamol overdose each year, and approximately 200 people die, most of whom presented late or did not receive antidote, N-acetylcysteine, within 12 hours. Nonsteriodal anti-inflammatory drugs (NSAIDs) have greater adverse effects in therapeutic use than paracetamol but also have a lower incidence of severe features or death in overdose. There is no antidote available for NSAID poisoning. Aspirin carries both significant adverse effects in therapeutic dose and a substantial risk in overdose, for which there is no antidote. Its risk benefit profile is probably the poorest of all analgesics currently available over-the-counter (OTC); this is reflected in current trends both in analgesic use and overdose figures. Although a number of options to reduce deaths from poisoning by OTC analgesics have been considered, few are practical, and all must take account of the public health benefits provided by these drugs. A perspective should be retained that the vast majority of the population in Australia, the United States, the United Kingdom, and Denmark derive therapeutic benefit from OTC analgesics and do not take them in overdose. The majority of those who do take overdoses come to little or no harm. Management of serious poisoning by paracetamol, aspirin, or NSAIDs remains a medical challenge. PMID- 11941386 TI - [Effect of erythropoietin 3-enhancer on proliferation of pulmonary artery smooth muscle cells induced by hypoxia]. AB - The effects of erythropoietin (EPO)3 -enhancer on endothelium-dependent and endothelium-independent proliferation caused by hypoxia in cultured porcine pulmonary artery smooth muscle cells (PASMCs) were investigated with MTT test, H(3)-TdR incorporation and flow-cytometry. The results showed that (1) PASMCs exposed to hypoxia for 24 h proliferated significantly, which was suppressed by pretransfecting wild type EPO3 -enhancer fragments into PASMCs, but not by protransfecting mutant fragments; and (2) the conditioned medium of pulmonary artery endothelial cells (PAECs) exposed to hypoxia for 24 h promoted the proliferation of PASMCs. This effect was abolished when wild type EPO3 -enhancer fragments were transfected, but it persisted when mutant fragments were transfected. The results suggest that (1) the conditioned medium of hypoxic PAECs induces proliferation of PASMCs. This may be because that not only PAECs are sensitive to hypoxia, but also PASMCs respond to hypoxia directly; and the hypoxic responses of both endothelial cells (ECs) and smooth muscle cells (SMCs) can be inhibited by exogenous EPO3 -enhancer fragments. (2) Since there is a HIF 1 binding site in EPO3 -enhancer, there may be a common pathway for HIF-1 hypoxia signal transduction in hypoxic responses of ECs and SMCs. PMID- 11941385 TI - Areas of emerging interest in analgesia: cardiovascular complications. AB - Many people obtain symptomatic relief from acute, chronic, or recurring pain conditions by using an over-the-counter analgesic. As with the use of any drug, this involves achieving the appropriate balance between potential benefit and risk of harm. The adverse effects of aspirin and nonsteroidal anti-inflammatory drugs (NSAIDs) in the gastrointestinal (GI) tract are widely appreciated. On the basis of their pharmacology, however, these drugs also have the potential for causing adverse effects in the cardiovascular system. This is particularly the case in certain overlapping populations (eg, the elderly or those with cardiac failure, hypertension, or renal impairment). And the size of the exposed populations and the fact they comprise people likely to require pain management because of concomitant illnesses make the cardiovascular implications of analgesic use potentially a more serious issue for public health than the more widely recognized GI complications of aspirin and NSAID use. This article discusses the impact on the cardiovascular system of different classes of analgesics (NSAIDs, the new cyclooxygenase-2-selective inhibitors [CSIs], and paracetamol) in terms of cardiac function, thrombotic and cardioprotective potential, and hypertension. It identifies patients at risk for analgesic-related cardiovascular adverse events, and considers their options for managing mild-to moderate pain. Unlike that of the NSAIDs and CSIs, the pharmacology of paracetamol provides no signal for risk of cardiovascular adverse events, and paracetamol should, therefore, be considered as first-line therapy in patients with cardiovascular disease. PMID- 11941387 TI - A study on the electrophysiological heterogeneity of rabbit ventricular myocytes the effect of ischemia on action potentials and potassium currents. AB - With the whole-cell variant patch-clamp technique, action potentials (AP) and outward potassium currents of rabbit ventricular myocytes isolated from subendocardium and subepicardium were recorded and their changes were observed under normal and ischemia conditions. The results showed that (1) under normal condition, there were differences in the AP figures between ventricular subendocardial and subepicardial myocytes. Action potentials recorded from subepicardial myocytes had shorter action potential duration (APD) and a notch between phases 1 and 2, compared with those of subendocardial myocytes. The resting potential had no significant difference between these two populations of the action myocytes; (2) under ischemia condition, the notch of action potentials of subepicardial myocytes disappeared and the APD was shortened even more, compared with that of subendocardial myocytes; (3) under normal condition, the density of steady-state outward potassium currents of subepicardial myocytes was significantly greater than that of subendocardial myocytes; (4) under ischemia condition, the increase of steady-state outward potassium currents of subepicardial myocytes was greater than that of subendocardial myocytes. Glybenclamide could partly reverse the above changes. It is suggested that the increase of steady-state outward potassium currents during ischemia is mainly due to the opening of I(K-ATP) channels as a result of the deficiency of intracellular ATP caused by ischemia. PMID- 11941388 TI - [MKP-1 regulates the cardiomyocyte hypertrophic responses induced by angiotensin II]. AB - The aim of this study was to determine the regulation by MKP-1 of MAPK activity and protein expression in cardiomyocyte hypertrophic response induced by Ang II. Neonatal rat cardiomyocyte hypertrophic response was assayed by cell surface area, protein synthesis rate and protein content. MAPK activity was determined by an in-gel kinase assay. Protein expression of MAPK and MKP-1 were detected by Western blotting. The results are as follows. (1) Ang II induced promotion of (3)H-leucine incorporation and increase in cell protein content and cell surface area in a dose-dependent manner. Pretreatment with a selective AT(1) receptor antagonist CV11974 or a specific MEK inhibitor PD098059, cardiomyocyte hypertrophic response induced by Ang II was inhibited by 85% and 32.5%, respectively. (2) After pretreatment with PD098059 or CV11974, AngII-induced increases in p44MAPK and p42MAPK protein expression and enzyme activity (expressed by gamma-(32)P-ATP incorporation) were all inhibited obviously. (3) With treatment of myocytes by Ang II for 5 min, MAPK activity determined by p44MAPK and p42MAPK protein expression began to increase, while MKP-1 protein expression was detected within 30 min and lasted more than 2 h following treatment with Ang II. (4) Pretreatment of cardiomyocytes with actinomycin D (3 microgram/ml) for 30 min inhibited MKP-1 protein expression, while p44MAPK and p42MAPK protein expression was still detected 120 min after Ang II treatment. The above results demonstrate that activation of MAPK plays an important role in Ang II-induced cardiomyocyte hypertrophic response in neonatal rat cardiomyocytes through MKP-1 mediated inactivation of p44MAPK and p42MAPK.cardiomyocyte hypertrophic response in neonatal rat cardiomyocytes through MKP-1 mediated inactivation of p44MAPK and p42MAPK. PMID- 11941389 TI - [Angiotensin II participates in stress-induced high blood pressure via stimulating hypothalamic vasopressin synthesis and release]. AB - Experiments were carried out in male Sprague-Dawley rats. The animals were randomly divided into three groups: control, stressed and stress + captopril. Stress stimulations were composed of repeated electric foot-shock combined with noise, twice one day (2 h each session) for 15 consecutive days. Animals in the stress+captopril group were administered with captopril (50 mg/kg.d) intraperitoneally. The results showed that at the end of the 15-day experiment the systolic pressure of the tail artery in stressed rats was significantly higher than that of the control rats, i.e., 19.75+/ C1.0 kPa (n=8, P<0.05) versus 16.32+/ C0.55 kPa (n=7); the vasopressin (AVP) mRNA level in the hypothalamus of the stressed rats also increased significantly compared with that of the control rats, i.e., 12990.33+/ C1533.58 (n=6, P<0.001) versus 7332.66+/ C522.65 (n=6). However, in the stress + captopril rats, both the tail artery systolic pressure and hypothalamic AVP mRNA level were significantly higher than those of the control rats, but lower than those of the stressed rats. In the control rats, no significant change in mean blood pressure (MBP) was observed after intracerebroventricular (icv) injection of 0.3 microgram of d(CH(2))(5)Tyr(Me)AVP, a selective AVP V(1) receptor antagonist; however, a decrease in MBP was observed in both stressed and stress+captopril rats (P<0.05), but the decrease in stress+captopril rats was more obvious than that of the stressed rats after icv a same dose of d(CH(2))(5)Tyr(Me)AVP. These results indicate that the endogenous renin-angiotensin system participates in the mechanism of the stress-induced high blood pressure in rats, and that the effect of Ang II is mediated mainly by stimulating hypothalamic AVP synthesis and release, which in turn result in an increase in blood pressure by acting on the central V (1) receptors. PMID- 11941390 TI - Intermittent hypoxia exposure prevents mtDNA deletion and mitochondrial structure damage produced by ischemia/reperfusion injury. AB - In the present study, polymerase chain reaction (PCR) was conducted to determine mtDNA(4834) deletion, and myocardial ultrastructure was visualized by electron microscope to see whether intermittent hypoxia (high altitude) adaptation exerts some action on mitochondria against ischemia/reperfusion injury. Myocardial ischemia/reperfusion in isolated perfused rat hearts induced severe damage to the ultrastructure of myocardial mitochondria and mtDNA4834 deletion down to 87.5% of normoxia rats. After the rats were exposed to intermittent hypoxia (5000 m; 6 h/d for 28 d), the myocardial structure was well reserved and mtDNA(4834) deletion dropped to 28.57%of control (P<0.05). It is suggested that intermittent hypoxia adaptation prevents mtDNA deletion, and preserves normal structure of mitochondria, which would be beneficial to the maintenance of normal mitochondrial function, and increases tolerance of myocardium against ischemia/reperfusion injury. PMID- 11941391 TI - [Modulation of GABA-activated currents by oxytocin in rat dorsal root ganglion neurons]. AB - Experiments were performed on freshly isolated dorsal root ganglion (DRG) neurons of rat. GABA(A)-activated currents were recorded using the whole-cell patch clamp technique. The majority of the neurons (48/52, 90.5%) were sensitive to GABA (10( 6)~10( 3) mol/L). Application of oxytocin (OT) induced outward membrane responses in 51.3% (20/39) of the neurons, no apparent responses in 43.6% (17/39) and inward responses in 5.1% (2/39). 10( 12), 10( 11), 10( 10) and 10( 9) mol/L OT increased 10( 4) mol/L GABA-activated currents to 24.1+/-7.6% (n=6), 33.4+/-6.9% (n=9), 40.2+/-6.5% (n=13) and 67.2+/-14.8% (n=5), respectively. After preapplication of OT, the Kd value for GABA(A)-activated currents decreased, while the response obtained at the maximum concentration increased. The results suggest that the enhancement of GABA-activated currents by OT may suppress primary sensory transmission by potentiating pre-synaptic inhibition of GABA. PMID- 11941392 TI - [Effect of scald on gene transcription and content of endothelin-1 in supraoptic nucleus of rat hypothalamus]. AB - Endothelin-1 (ET-1) gene transcription and endothelin-1-immunoreactivity (ET-1 ir) in the supraoptic nucleus (SON) of rat hypothalamus were respectively observed by in situ hybridization and immunohistochemistry after scald. Intensity of ET-1 mRNA and endothelin-1-immunoreactivity (ET-1-ir) was quantified by image analysis. Compared with the control (sham scald) group, no significant change in the intensity of ET-1 mRNA positive hybridization signals in SON was found 15 min post-scald, while there was a 35.1% increase in the positive hybridization signal intensity 60 min post-scald (P<0.05) and a 62.4% increase 180 min post-scald (P<0.01). The content of ET-1-ir in SON decreased significantly to 8.5% of the control 15 min post-scald (P<0.01), and gradually recovered to 31.5% and 52.4% of the control 60 min and 180 min post-scald respectively, though still significantly lower than the control (P<0.01). Pre- and post-scald ET-1 gene transcription in rat hypothalamus was also measured by Northern blot hybridization. No significant difference in the quantity of ET-1 mRNA was found between 15 min post-scald data and those of the control. The quantity increased to a significantly higher level 60 min post-scald (P<0.05) and further increased to 2.5 creased to 2.5 fold of the control 180 min post-scald (P<0.05). In addition, the Northern blot hybridization showed that the post-scald size of ET-1 mRNA remained unchanged despite of the increase in quantity. In view of the neuroendocrine role of SON, the changes in ET-1 mRNA and ET-1-ir in SON resulting from scald suggest that ET-1 may play an important role in neuroendocrine reactions following scald. PMID- 11941393 TI - [The role of theophylline in the improvement of scop-induced learning and memory impairment]. AB - Adenosine and ACh contents in hippocampus, striatum region around the nucleus of basal meynert (NBM) and the frontal and temporal parts of rat cortex were measured by a high performance liquid chromotography-electronic detector (HPLC ECD). Adenosine content in 18~20 month-old rats was significantly higher than that of 3~6 month-old rats, while ACh content in 18~20 month-old rats was lower than that of 3~6 month-old ones. Learning and memory impairment models were established by induction of SD rats with scopolamine (0.5 mg/kg, ip), which were injected with an adenosine receptor antagonist theophylline (1.0 mg/5 microliter, icv), and the step-down test was performed. The error frequency in the step-down test decreased significantly (P<<0.05), and the retention time was longer than that of control. Furthermore, ACh content increased in all the three different brain regions. The above results suggest that adenosine may play an important role in senile learning and memory impairment, and theophylline may improve scopolamine-induced learning and memory impairment by increasing ACh content. In consequence, theophylline could be a promising drug for ameliorating senile learning and memory impairment. PMID- 11941394 TI - [Relationship between the sensitivity to tetraethylammonium and firing patterns of injured dorsal root ganglion neurons]. AB - In order to study the relationship between the firing patterns of injured dorsal root ganglion (DRG) neurons of rat and their sensitivity to tetraethylammonium (TEA), spontaneous activities from single fibers of chronically compressed DRG neurons were recorded. Two patterns of spontaneous activity were found: periodic pattern and non-periodic pattern. These two types of neurons showed different sensitivity to TEA with 27.3% in periodic and 93.2% in non-periodic firing neurons responding to TEA (2 mmol/L) (P<0.01). The responses to TEA of different concentrations were greater in the non-periodic firing neurons than in the periodic firing ones (P<0.01). The responses to TEA became apparent in 5 neurons after their periodic firing pattern was transformed into non-periodic firing pattern through an increase in Ca(2+). The non-periodic interburst interval of the bursting discharge decreased significantly while the periodic burst interspike interval had no response to TEA. The unstable period orbits were found in bursting discharge, indicating deterministic chaos during bursting discharge. It is concluded that non-periodic firing neurons with chaos are more sensitive to TEA than periodic firing neurons. PMID- 11941395 TI - [Pituitary adenylate cyclase activating polypeptide ameliorates the damage and inhibits the increase of intracellular calcium concentration in cultured hippocampal neurons induced by glutamate]. AB - Hippocampal neurons cultured from 7 to 9 d in vitro were used to observe the effect of glutamate. Treatment of glutamate for 24 h greatly decreased neuronal survival and pretreatment with pituitary adenylate cyclase activating polypeptide (PACAP) significantly attenuated hippocampal neuron death induced by glutamate. Moreover, glutamate dose-dependently increased the intracellular calcium concentration in cultured hippocampal neurons, while PACAP inhibited the increase of intracellular calcium concentration induced by glutamate. PACAP 6-38, a specific PACAP type I receptor antagonist, completely inhibited the amelioration of glutamate induced death and the decrease of intracellular calcium concentration induced by PACAP in cultured hippocampal neurons. The data suggest that PACAP has a neuroprotective effect on the hippocampal neuronal damage induced by glutamate, which is related to an inhibition of glutamate-induced increase of intracellular calcium concentration and mediated by PACAP type I receptor. PMID- 11941396 TI - [Effect of sodium nitroprusside on the spontaneous and induced responses of rat spinal cord dorsal horn neurons]. AB - The present study was performed on anesthetized and paralyzed Wistar rats. Extracellular recordings were made from the lumbar enlargement of the spinal cord using carbon filament electrodes. NO donor sodium nitroprusside (SNP) was locally microdialyzed to the spinal cord, and effects of NO on spontaneous response and the responses induced by mechanical stimulation of the hind-foot were observed. After dialyzing for 10~20 min, SNP at a concentration of 1 micromol/L increased the induced responses to innocuous mechanical stimulation and decreased those to noxoius mechanical stimulation. The induced responses to both innocuous and noxious mechanical stimulations were all decreased after dialyzing the same dose of SNP for 20~30 min. The decrease was shown within 7~15 min dialysis when SNP was used at a concentration of 20 micromol/L. SNP at both concentrations of 1 and 20 micromol/L increased spontaneous spikes of the recorded spinal units. These effects were shown preferentially in deep units. The data suggest that in the deeper neurons of the spinal cord NO of different concentrations has different effects on transmission of nociceptive and nonnociceptive signals induced by mechanical stimulation, and No is involved in the antinociception in spinal neurons and facilitates their spontaneous activity. PMID- 11941397 TI - [Spatial frequency tuning characteristics of cat primary visual cortex at different topological locations by optical imaging]. AB - Using optical imaging based on intrinsic signals, we studied spatial frequency tuning characteristics of cat primary visual cortex at different visual topological locations. We found that the areas representing the peripheral visual field had null or very weak responses to high spatial frequency grating stimuli, whereas the areas representing the central visual field responded to the stimuli of a wide range of spatial frequencies with greater responses to high spatial frequencies. The more centered the corresponding visual field of the cortical area, the higher the cutoff spatial frequency. The spatial frequency tuning curves also tended to have a smooth shift along the cortical surface. The results above demonstrate that spatial frequency tuning characteristics of the primary visual cortex change according to different visual topological locations of the cortical areas, in addition to the existence of spatial frequency columns. It is suggested that the distribution of spatial frequency columns may be related to visual topology. PMID- 11941398 TI - [Effect of central administration of motilin on the activity of gastric-related neurons in brain stem and gastric motility of rats]. AB - To investigate the effect of microinjection of motilin into ventral medail hypothalamus (VMH) or intracerebroventricle (icv) on gastric motility, extracellular unit responses of dorsal vagal complex (DVC) in anesthetized rats and gastric motility of conscious rats were recorded. The results are as follows. (1) The firing rate of gastric distention related neurons in DVC was modified. (2) VMH and icv administration of motilin induced obvious enhancement in gastric motility. (3) The effect of motilin on gastric motility was abolished by vagotomy. These results imply that central motilin appears to work as an excitatory neuromodulator in the regulation of gastric motility through VMH-DVC vagal nerve axis. PMID- 11941399 TI - Volume-activated chloride current in pigmented ciliary epithelial cells. AB - To characterize the volume-activated Cl(-) current in bovine pigmented ciliary epithelial (PCE) cells, the whole-cell patch clamp technique was employed. Exposure to a hypotonic solution induced a volume-activated Cl(-) current. The activation of the current depended on intracellular ATP. The current showed a characteristic outward rectification. There was no, or negligible, time-dependent inactivation of the current. The current-voltage relationship showed that the reversal potential of the hypotonic-activated current ( 6.3 0.5 mV) was close to the calculated equilibrium potential for Cl(-) (ECl=0 mV). Extracellular ATP inhibited both outward and inward currents, but the inhibition of outward currents was larger than that of inward currents (92% vs 74%, P<0.01). The current was also blocked by extracellular application of tamoxifen, a Cl( ) channel inhibitor, the effects of which were almost equal for both outward and inward currents (85% vs 87%, P>0.05). The properties of this current are quite similar to those of the volume-activated Cl(-) current associated with P glycoprotein in other cell types, suggesting that P-glycoprotein may be involved in the activation of the volume-activated Cl(-) current in PCE cells. PMID- 11941400 TI - [Inhibition of transmembrane K(+) currents in mammalian ventricular myocytes by antiarrhythmic agent RP62719]. AB - The effect of RP62719 on the inward rectifier K(+) current (I(K1)),transient outward K(+) current (I(to)) and delayed outward K(+) current (I(K)) in isolated cardiac myocytes was determined using the whole cell patch clamp technique in guinea pig and dog. RP62719 decreased I(K1) with an inhibitory concentration 50 (IC(50) ) of 5.0+/-1.0 micromol/L at -100 mV in guinea pig ventricular cells. In dog ventricular myocytes, RP62719 inhibited Ito by 84+/-4.4% with an IC(50) of 1.2+/-0.51 micromol/L at +40 mV. In guinea pig ventricular cells, RP62719 decreased I(K): I(Kstep) by 50.0+/-8.3%%and I(Ktail) by 56.0+/-4.9% at +40 mV, respectively. RP62719 inhibited I(Kstep) with an IC(50) of 4.2+/-0.8 micromol/L and I(Ktail) with an IC(50) of 3.3+/-0.75 micromol/L. Thus it is suggested that the ionic mechanism of antiarrhymic effect by RP62719 may be due to its inhibition of I(K1),I(to) and I(K). PMID- 11941401 TI - [The oblique effect revealed by optical imaging in primary visual cortex of cats]. AB - The oblique effect is a ubiquitous visual psychological effect. To explore its underlying neural basis, we quantitatively analyzed the proportion and response amplitude of the cardinal preferred areas and the oblique preferred areas in a fairly large region of the primary visual cortex of cats, using optical imaging based on intrinsic signals. The results show that cardinal preferred areas were larger than oblique preferred areas, with a mean difference of 4.7%. Overall, the responses evoked by cardinal stimuli were generally greater than those by oblique stimuli. The present work provides an explanation for the differences in electrophysiological results reported for this issue, and gives a new insight into the neural basis of the oblique effect. PMID- 11941402 TI - Changes in heart rate, blood pressure and renal sympathetic nerve activity induced by microinjection of capsaicin into area postrema in rats. AB - The effects of capsaicin microinjection into area postrema (AP) on mean arterial pressure (MAP), heart rate (HR) and renal sympathetic nerve activity (RSNA) were investigated in 36 anesthetized Sprague-Dawley rats. The results obtained are as follows. (1) Following microinjection of capsaicin (10 micromol/L, 50 nl) into the AP, MAP, HR and RSNA were significantly increased from 12.34+/-0.53 kPa, 328.52+/-7.54 bpm and 100+/-0% to 15.17+/-0.25 kPa (P<0.001), 354.81+/-8.54 bpm (P<0.001) and 156.95+/-7.57% (P<0.001), respectively. (2) Ruthenium red (RR, 100 mmol/L, 0.2 ml, iv), a capsaicin receptor antagonist, significantly inhibited these effects of capsaicin. (3) Pretreatment with a NMDA receptor antagonist MK 801 (500 microgram/kg, 0.2 ml, iv) also reduced these effects of capsaicin. The above results indicate that microinjection of capsaicin into AP induces excitatory effects on MAP, HR and RSNA, which are mediated by capsaicin receptors with glutamate involvement. PMID- 11941403 TI - [Analysis of P/N leak subtraction of pClamp acquisition software]. AB - Using blind whole cell recording technique on rat hippocampal slices, the function and mechanisms of P/N leak subtraction of Clampex program in pClamp acquisition software were studied. The ways of picking up sub-pulse current and properly selecting parameters, including time course, polarization, sub-holding potential, numbers and location of P/N sub-pulse were analyzed in detail. Our results indicate that voltage-gated ion channel currents recorded by Clampex will be more accurate and reproducible by properly using P/N leak subtraction. PMID- 11941404 TI - [17beta -estradiol inhibits carotid sinus baroreflex in male rats]. AB - By perfusing isolated carotid sinus, the effect of 17beta-estradiol (E(2)) on carotid sinus baroreflex was observed in anesthetized male rats. The results obtained are as follows. (1) By perfusing with E(2) (10 micromol/L), the functional curve of baroreflex was shifted to the right and upward, with a peak slope (PS) decrease from 0.49+/-0.03 to 0.25+/-0.01 (P<0.01) and a reflex decrease in mean arterial pressure (reflex decrease, RD) from 7.37+/-0.42 kPa to 3.49+/-0.20 kPa (P<0.001), while the threshold pressure (TP) and saturation pressure (SP) were significantly increased from 9.52+/-0.68 kPa to 13.3+/-0.11 kPa (P<0.001) and 24.53+/-0.48 kPa to 27.52+/-0.20 kPa (P<0.01) respectively. Among the functional parameters of carotid baroreflex, the changes of RD, PS, TP and SP were dose-dependent. (2) Pretreatment with different doses of tamoxifen (1, 5, 10, 30 micromol/L), an inhibitor of estrogen receptor, did not block the effect of E(2) on carotid baroreflex. (3) Preperfusion with an inhibitor of NO synthase L-NAME (100 micromol/L) could completely abolish the effect of E(2) on carotid baroreflex. It is concluded that the inhibitory effect of E(2)on carotid sinus baroreflex may be mediated by NO release from endothelial cells, but not by a genomic mechanism. PMID- 11941405 TI - Nitric oxide inhibits the expression of proto-oncogene c-fos induced by angiotensin II and endothelin-1 in cardiomyocytes. AB - The effect of nitric oxide (NO) on the hypertrophic response and the proto oncogene c-fos expression induced by angiotensin II (AII) or endothelin-1 (ET-1) was investigated in the primary culture of neonatal rat cardiomyocytes. Total protein content of cardiomyocytes (used as the index of cardiac myocyte hypertrophy) was determined by the Bradford method. The proto-oncogene c-fos expression was assessed using reverse transcription-polymerase chain reaction (RT PCR) standardized with glyceraldehyde-3-phosphate dehydrogenase (GAPDH). RT-PCR was performed in a single tube using gene-specific primers and the SuperScript One-Step RT-PCR System. Total protein content of cardiomyocytes increased significantly on day 5 after A II treatment or on day 3 after ET-1 treatment and the increased protein content was inhibited by SNP (NO donor). A II, ET-1 and PMA (protein kinase C activator) induced the c-fos gene expression of cardiomyocytes, while L-arginine inhibited it. The L-arginine effect was blocked by L-NAME (NOS inhibitor). SNP inhibited the c-fos gene expression of cardiomyocytes induced by A II,ET-1 or PMA as well. These results suggest that NO can inhibit the hypertrophic response and the proto-oncogene c-fos expression of cardiomyocytes induced by A II or ET-1 and the cross-link may be located at the site of protein kinase C. PMID- 11941406 TI - [Content and activity of the focal adhesion kinase in cultured vascular smooth muscle cells enhanced by urotensin II]. AB - Urotensin II (U II) is the most potent vasoconstrictor identified in vivo, which plays an important role in the smooth muscle cell proliferation in atherosclerosis. All available information suggests that focal adhesion kinase (FAK) is at the crossroads of multiple signaling pathways and is essential for cell proliferation. But the effect of U II on the FAK mediated signal transduction pathway is unclear. In this study, FAK content and tyrosine phosphorylation were assessed by Western blot and immunoprecipitation in cultured rat vascular smooth muscle cells. Increased protein tyrosine phosphorylation was observed within 5 min of U II 10(-7) (mol/L) addition and was maximal by 30 min, while FAK protein content showed no change during the first 30 min but it increased at 2 h reaching a plateau by 4 h, and decreased after 6 h. In addition, the elevated phosphorylation of FAK was detected upon U II stimulation at 10(-8) mol/L, being maximal at 10(-7) mol/L, but decreased at 10(-6) mol/L. Treatment of the cells with cytochalasin B (50 micromol/L), which disrupted the organization of cytoskeleton, had no influence on the increased FAK tyrosine phosphorylation in response to U sti II mulation. In order to study the relationship between FAK and mitogen-activated protein kinase, calmodulin and protein kinase C, selective inhibitors PD98059 (50 micromol/L), W7 (50 micromol/L) and H7 (50 micromol/L) were added following U II treatment. Neither PD98059 nor W7 influenced the increased FAK tyrosine phosphorylation, but H7 further increased it. These findings indicate that FAK activation is independent of the integrity of cytoskeleton and closely related to protein kinase C, but had no relation with mitogen activated protein kinase and calmodulin. PMID- 11941407 TI - [Alteration of the expression of rat cardiac Galphaq/11 and Gialpha2 proteins during endothelin-1 pre-treatment]. AB - The present study was undertaken to explore the mechanism of G protein-mediated signal transduction pathway during endothelin-1 (ET-1) pre-treatment and ischemic preconditioning (IP). Rats were divided into four groups: ET-1, IP, ischaemia reperfusion (IR) and control groups. ET-1 pre-treatment model was prepared by administrating 0.5 nmol/(L.kg) ET-1 into rat left ventricle, whereas IP model was prepared by ligating the left coronary artery for 5 min followed by 30 min reperfusion. All the animals were subjected to 60 min regional ischaemia and 30 min reperfusion alternately and then parameters of ventricular arrhythmia and expression of cardiac Galphaq/11 and Gialpha2 were measured. The results showed that the scores of ventricular arrhythmia decreased significantly in both ET-1 and IP treated groups as compared with IR group. In comparison with control group, Galphaq/11 increased by 77.8% (P<0.05) and 110.6% (P<0.01) in IP and ET-1 group respectively. Gialpha2 showed no significant difference in IP group, while it decreased by 31.0% (P<0.01) in ET-1 group. In conclusion, activation of G alphaq/11 may be related to the protecting mechanism of ET-1 pre-treatment and IP, whereas Gialpha2 may only play a role in ET-1 pre-treatment. PMID- 11941408 TI - [Role of calcium in the mechanism underlying the inhibitory effect of streptomycin on carotid sinus baroreflex in rats]. AB - The effect of streptomycin (SM) on carotid baroreflex was examined in 23 anesthetized rats with isolated carotid sinus perfusion. The results obtained are as follows. (1) In response to perfusion with SM (200 micromol/L), the functional curve of carotid baroreflex was shifted to the right and upward with a decrease of peak slope (PS) and a reflex decrease in mean arterial pressure (RD)(P<0.01), indicating an inhibitory effect of SM on carotid sinus baroreflex. (2) By perfusing the isolated carotid sinus with high Ca(2+) solution (4 mmol/L), the inhibitory effect of SM on carotid baroreflex was partially eliminated. The functional curve of SM was shifted to the left and downward with PS increasing from 0.27+/-0.04 kPa to 0.37+/-0.02 kPa (P<0.01) and RD was enhanced from 4.32+/ 0.14 kPa to 6.18+/-0.17 kPa (P<0.01). On the other hand, the threshold pressure (TP) and saturation pressure (SP) were significantly decreased from 10.29+/-0.29 kPa to 9.98+/-0.33 kPa (P<0.05) and from 27.26+/-0.42 kPa to 25.22+/-0.38 kPa (P<0.05), respectively. (3) By pretreatment with Bay K 8644 (500 nmol/L), an agonist of calcium channels, the effect of SM on carotid baroreflex was completely abolished. (4) Exposure of the carotid sinus to SM following pretreatment with charybdotoxin (ChTX,100 nmol/L), a blocker of the Ca(2+) activated K(+) channel (KCa), still inhibited the baroreflex. These results suggest that the inhibitory action of SM on carotid baroreflex may be mediated by suppressing Ca(2+) influx. PMID- 11941409 TI - [Effect of acetylcholine on the discharge of presympathetic neurons of the rostral ventrolateral medulla of rats]. AB - Effect of acetylcholine (ACh) on presympathetic neurons of the rostral ventrolateral medulla (RVLM) was studied by extracellular recording technique and microiontophoresis in urethane-anaesthetized rats. Spontaneous discharges of 35 presympathetic neurons of RVLM were recorded. All the neurons showed increased discharge by microiontophoresis of ACh dose- dependently with the increase of iontophoretic current. In 22 units microiontophoresis of atropine decreased the baseline discharge rate and completely blocked the excitatory effect of ACh on these neurons, while in the remaining 7 and 6 neurons microiontophoresis of d tubocurarine (d-TC) and hexamethonium (C(6)) respectively had no effect and did not block the action of ACh. The above finding suggests that presympathetic neurons of RVLM receive a tonic excitatory cholinergic input mediated by M receptors. PMID- 11941410 TI - [Cardiovascular effects of lateral intracerebroventricular injection of L securinine]. AB - To investigate cardiovascular effects of changed GABAergic tonic activation in periventricular forebrain, arterial pressure (AP), heart rate (HR) and renal sympathetic nerve discharge (RSND) were recorded in anesthetized rats when L securinine (L-Sec) was administered into the lateral intracerebroventricle. L-Sec elicited dose-dependent increases in RSND, AP and HR, which were much weaker than those of bicuculline. L-Sec antagonized the sympatho-inhibitory and depressor effects evoked by both muscimol and baclofen. These results indicate that GABAergic inhibition originating from periventricular forebrain may suppress tonically sympathetic outflow to cardiovascular system which is disinhibited by L Sec, and L-Sec is likely an unselective GABA receptor antagonist. PMID- 11941411 TI - [17beta -estradiol induced nitric oxide release in vascular endothelial cells]. AB - Bovine aortic endothelial cells (BAECs) were used to study the effect of 17beta estradiol (E(2)) on nitric oxide (NO) release, nitric oxide synthase (eNOS) mRNA expression and intracellular free calcium con~cen~tration ([Ca(2+)](I)) and modulation of the effect of E(2) by estrogen receptor (ER) antagonist tamoxifen and NOS inhibitor L-NAME. E(2) (10(-12) 10(-8) mol/L) induced NO release of BAECs in a concentration-dependent manner and the abundant expression of eNOS mRNA in BAECs increased obviously after treatment with E(2) (10(-8)mol/L) for 48 h. These effects were evidently inhibited by tamoxifen (10(-7)mol/L) and L-NAME (10(-6) mol/L). Furthermore treatment with E(2) (10(-8) mol/L) for 48 h significantly increased the resting [Ca(2+)](I) and the rise of [Ca(2+)](I) induced by ATP in BAECs. These results suggest that E(2)-induced NO release and eNOS mRNA expression in BAECs may be mediated by ER and related to calcium mobilization. PMID- 11941412 TI - [Role of pp60c-src in mitogen-activated protein kinase activation of vascular smooth muscle cells]. AB - The effect of pp60c-src on mitogen-activated protein kinase (MAPK) activation induced by angiotensin II (Ang II) in vascular smooth muscle cells (VSMCs) was observed to explore the signal transduction process of VSMCs proliferation mediated by Ang II. Following transfection of the synthetic antisense c-src oligodeoxynucleotides (ODNs) wrapped in lipofectin into cultured rat VSMCs, pp60c src content markedly decreased, pp60c-src kinase activity was only 14.4% of control, while MAPK activity was not significantly changed (P>0.05). When these transfected VSMCs were further stimulated by Ang II, the increase in MAPK activity was only 1.6% of control. These data suggest that pp60c-src is needed for MAPK activation in VSMCs stimulated by Ang II, and pp60c-src may be an important signaling molecule in abnormal proliferation of VSMCs. PMID- 11941413 TI - [Effect of interleukin 2 on the expression of estrogen receptors in the anterior pituitary of rats]. AB - Using primary serum-free cell culture combined with immunocytochemistry and semi quantified RT-PCR methods, we observe the modulation of estrogen receptor alpha (ERalpha) and beta (ERbeta) by interleukin-2 (IL-2) in rat anterior pituitary. The results show that IL-2 up-regulates the level of ERalpha protein and the expression of ERalpha mRNA, but down-regulates those of ERbeta. The density per cell of ERalpha-immunoreactive (ir) cells increases from 48.740 4.567 to 81.188+/ 6.619, whereas that of ERbeta -ir cells decreases from 102.560+/-6.250 to 72.718+/-7.623 after rhIL-2 (10 microgram/L) incubation for 48 h. In parallel with these changes, the ratio of ERalpha/ beta -actin mRNA increases from 0.1511 to 0.4334, and ERbeta /beta -actin mRNA declines from 0.3822 to 0.1528. It is likely that IL-2 has direct regulatory effect on ER in anterior pituitary. PMID- 11941414 TI - [Effects of muscle spindle afferents on the regulation of respiration in rabbit]. AB - Effect of muscle spindle afferent invoked by intraarterial injection of succinylcholine (Sch) on respiration was assessed by changes in unit discharge of respiration related neurons and phrenic nerve of 63 anesthetized, bilaterally vagotomized and artificially ventilated rabbits. The results are as follows. (1) Intra~arterial injection of Sch prolonged inspiratory duration (Ti) or decreased expiratory duration (Te) and augmented respiratory frequency (RF). (2) 66% (37/56) of the medullary inspiratory neurons were excited and 10/17 of the expiratory neurons were inhibited by intraarterial injection of Sch, reaching maximum respectively at 1 2 and 2 3 min after injection, while non-respiration related neurons were not affected. (3) This effect of Sch was suppressed or abolished when muscle spindles were destroyed by intramuscular injection of bupivacaine. PMID- 11941415 TI - [mRNA and protein expression of skeletal DHPR(alpha1) and RyRs in diaphragm muscle of rabbits]. AB - To detect mRNA and protein expression of skeletal dihydropridine receptor isoform alpha1 subunit and ryanodine receptor 1 and 3 in diaphragm muscle of rabbits, the coupling mode and characteristics of Ca(2+) release were explored. Reverse transcription PCR, in situ hybridization and immunohistochemical methods were employed. A higher level of mRNA and protein expression of DHPR(alpha1) and RyR(1), and a lower level of mRNA expression of RyR(3) were found. It is suggested that the calcium release unit may consist of skeletal DHPR isoform, RyR(1) and RyR(3), and there may be two kinds of Ca(2+) release mode via conformational changes in linked proteins and Ca(2+)-induced Ca(2+) release (CICR) in diaphragm muscle of rabbits. PMID- 11941416 TI - [Cholecystokinin-octapeptide alleviates tumor necrosis factor-alpha induced changes in rabbit pulmonary arterial reactivity and injuries of endothelium in vitro]. AB - To explore the mechanism underlying cholecystokinin-octapeptide (CCK-8) induced attenuation of pulmonary arterial hypertension (PAH) in endotoxic shock, the effects of CCK-8 on the changes in rabbit pulmonary arterial reactivity induced by tumor necrosis factor-alpha (TNF-alpha) were observed with the isolated arterial ring technique, and the ultrastructure of pulmonary arterial endothelium was observed under a scanning electron microscope. The contractile response to adrenoceptor agonist phenylephrine (PE), the endothelium-dependent relaxation response to acetylcholine (ACh) and the endothelium-independent relaxation response to sodium nitroprusside (SNP) were not affected by TNF-alpha (4000 U/ml) after incubation for 2 h, while, if the incubation time was prolonged to 7 or 14 h, the relaxation response of pulmonary artery to ACh was depressed significantly, which, however, could be reversed by concomitant exposure to CCK-8 (0.5 microgram/ml). Incubation of pulmonary artery with CCK-8 (0.5 microgram/ml) alone did not bring out any contractile responses. Moreover, CCK-8 (0.5 microgram/ml) alleviated the ultrastructural lesions induced by TNF-alpha (4000 U/ml). These results suggest that CCK could protect pulmonary arterial endothelium against the detrimental effects by TNF-alpha. PMID- 11941417 TI - [Relationship between memory enhancement induced by nitric oxide donor and brain ADP-ribosyltransferase in rats]. AB - In order to explore the signal transduction pathway of nitric oxide (NO) related to learning and memory, the effects of intracerebral ventricular injection (icv) of NO donor sodium nitroprusside (SNP) alone or along with ADP-ribosyltransferase (ADPRT) inhibitor nicotinamide (NIC) on learning and memory behavior were investigated in rats. Brain ADPRT activity was analyzed using high performance liquid chromatography. The results showed that SNP (0.36 microgram, icv) facilitated short-term and long-term memory in passive avoidance response and elevated learning and memory ability in active avoidance response. NIC (1.5 mg, icv) blocked the effect of SNP. Training in active avoidance task raised ADPRT activity in hippocampus. Treatment with SNP (icv) before training caused significant elevation of ADPRT activity in hippocampus and cerebral cortex. The results above support the view that ADPRT may be one of the target molecules of NO action. PMID- 11941418 TI - [Evoked responses of visceral nociceptive neurons in cat anterior cingulate gyrus revealed by intracellular recording techniques]. AB - With intracellular recording techniques, we investigated evoked responses and electrophysiological properties of neurons in anterior cingulate gyrus (ACG) to stimulating greater splanchnic nerve (GSN) in cats. Among 461 neurons, 176 were stimulus-relative and 285 stimulus-irrelative. According to the characteristics of the evoked responses, stimulus-relative neurons were classified into specific visceral nociceptive neurons (SVNNs, 64.77%), non-specific visceral nociceptive neurons (NSVNNs, 19.32%) and non-visceral nociceptive neurons (NVNNs, 15.91%). Modes of the evoked responses could be excitatory, inhibitory, or mixed ones. The results suggest that ACG may be one of the representative areas of the GSN afferent pathway, and there exist two kinds of VNNs in ACG, which may be differently involved in pain modulation. PMID- 11941419 TI - [Modulatory effects of 17beta-estradiol on the electrical activity of subfornical organ neurons]. AB - The effects of 17beta-estradiol (E(2) ) on electrical activity of neurons in subfornical organ (SFO) slices were examined using extracelluar recording technique. The results are as follows. (1) In 15 SFO units, a low dose of E(2) (0.1 nmol/L) applied into superfusate induced an increase in discharge rate from 3.21+/-0.37 to 6.79+/-0.71 Hz (P<0.001), whereas a high dose of E(2) (100 nmol/L ) caused a decrease in discharge rate from 3.44+/-0.40 to 1.44+/-0.36 Hz (P<0.01); (2) glutamate NMDA receptor antagonist MK-801 (50 pmol/L) blocked the excitatory effects induced by low dose of 17beta-estradiol in 7 units; (3) L arginine (L-arg, 1 mmol/L), a physiological precursor of NO, abolished the excitatory effects induced by low dose of 17beta-estradiol in 7 units; (4) application of N-omega-nitro-L-arginine methyl ester (L-NAME, 10 mmol/L), an inhibitor of NOS, blocked the inhibitory effects induced by high dose of 17beta estradiol in 6 units. The above results suggest that the estrogen exerts dual action on SFO neuron. E(2) at low dosage increases the discharge rate of SFO neuron, an effect which may be related to the activation of NMDA receptors, whereas E(2) at high dosage decreases the discharge rate, an effect which may be attributed to the activation of NOS with resultant production of NO. PMID- 11941424 TI - Meet the Editors. PMID- 11941420 TI - [Fibronectin or RGD peptide promotes nitric oxide synthesis of rabbit bronchial epithelial cells]. AB - To find out whether the extracellular matrix component fibronectin plays a regulatory and protective role in bronchial epithelial cells, the present study was undertaken to detect the NO released from primary cultured rabbit bronchial epithelial cells (BEC) and the activity of NO synthesase (NOS) in cells. Stress with ozone was taken as the positive control, and the effects of fibronectin (Fn) or its specific sequence Arg-Gly-Asp (RGD peptide) was observed. The results showed: (1) Ozone stress enhanced NO release from BEC, and treatment with Fn also promoted the NO production. The effect of Fn could be blocked by calmodulin inhibitor W(7). (2) Both Fn and RGD showed a dose-dependent promotion on NO release. (3) The NOS activity was significantly elevated in Fn treated group and the effect of Fn was abolished by W(7). A dose-response relation in NOS activity was observed in either Fn treated or RGD treated group. It is concluded that the binding of fibronectin or its specific sequence RGD peptide with integrins of BEC plays a role in upregulating the NOS activity and the NO release, and calmodulin may take part in the pathway of signal transduction. PMID- 11941425 TI - A tribute to Isao Karube and Rolf D. Schmid. PMID- 11941426 TI - Aldehydes 2001--an unusual conference. PMID- 11941427 TI - Determination of acetone, 2-butanone, diethyl ketone and BTX using HSCC-UV-IMS. AB - A combination of a custom-designed ion mobility spectrometer (IMS) with a UV ionization source and a high speed capillary column (HSCC) has been developed as an analytical device for the sensitive detection of volatile organic compounds (VOCs), e.g. 2-propanone (acetone), 2-butanone and 3-pentanone (diethyl ketone) in the gas phase. A fast separation of the three selected substances and benzene, toluene and m-xylene (BTX) - all of which occur in human breath - has been achieved within less than four minutes at a carrier gas flow rate of 4.5 mL x min(-1). Multi-dimensional correlations presented support the interpretation of the acquired spectra of mixtures. Method detection limits were 2.7 microg x L(-1) for acetone and 2-butanone and 3.0 microg x L(-1) for diethyl ketone in nitrogen, respectively. The assay linear dynamic range is 4-320 microg x L(-1). PMID- 11941428 TI - Monitoring apple flavor by use of quartz microbalances. AB - "Electronic noses", i.e. arrays of differently coated quartz microbalances (QMB), have been used for selective detection of, and discrimination between, volatile organic compounds (VOC) formed during the post-harvest ripening of apples. The flavor components to be differentiated are chemically rather similar carbonyl compounds, chiefly aldehydes and esters. Because their relative ratios change during the post-harvest ripening period, appropriately selected sensor-active layers lead to characteristic patterns of the sensor responses which can be analyzed via pattern-recognition methods. This enables qualitative and quantitative identification of individual components whereby the post-harvest ripening of apples and other fruits can be monitored. Different kinds of apple differ in type and concentration of individual carbonyl compounds. PMID- 11941429 TI - Method optimization for the determination of carbonyl compounds in disinfected water by DNPH derivatization and LC-ESI-MS-MS. AB - A method has been developed for quantitative determination of carbonyl disinfection by-products (DBP) from aqueous samples by derivatization with 2,4 dinitrophenylhydrazine combined with high-performance liquid chromatography (HPLC) and electrospray ionization (ESI) tandem mass spectrometry (MS-MS). The effect of excess of derivatization reagent and derivatization time, the effect of buffer and dry-gas temperature in the ESI process, and the effect of focus potential and collision energy in MS measurement are shown. Major fragment ions for compound identification on the basis of collision-induced dissociation (CID) mass spectra (MS) are given, as are common fragments for screening analyses by MS experiments such as the use of precursor ion scans. Detection limits in the microg x L(-1) range could be achieved by selected ion monitoring measurements without sample preconcentration. Solid-phase extraction improved the sensitivity by a factor of 25 to 250. The applicability of the method is illustrated by DBP analyses of samples from outdoor swimming pools after chlorination. Several carbonyl compounds, e.g. aldehydes, ketones, hydroxybenzaldehyde, and dicarbonyl compounds were identified. PMID- 11941430 TI - Qualitative and quantitative analysis of carbonyl compounds in ambient air samples by use of an HPLC-MS(n) method. AB - The suitability of HPLC combined with ion-trap mass spectrometry was studied for the determination of carbonyl-2,4-dintrophenylhydrazones in ambient air. MS quantification was based on two internal standards and atmospheric pressure chemical ionization in the negative-ion mode. Limits of detection for air samples of 750 L in the full-scan mode varied between 1 and 15 ng x m(-3) expressed as carbonyl. Limits of quantification were approximately a factor of three higher. This is sufficient for background regions. For sample volumes of 750 L air the instrument response was linear from 10 ng x m(-3) to 800 microg x m(-3) for carbonyls and from 3 ng x m(-3) to 250 ng x m(-3) for dicarbonyls. Besides complete method validation, quantitative results for six air samples from four background sampling sites in North and Central Europe were compared with those obtained by use of HPLC-UV. Thirty-six carbonyl compounds could be identified and twenty-four were quantified. Values for major compounds, i.e. those present at levels well above the UV detection limits (9 to 18 ng x m(-3)), deviated by less than 20%. PMID- 11941431 TI - HPLC-MS determination of the oxidation products of the reaction between alpha- and beta-pinene and OH radicals. AB - Biogenic non-methane hydrocarbons such as isoprene, alpha-pinene, and beta pinene, are emitted by forests in very large quantities. To evaluate the role of alpha- and beta-pinene and their contribution to the global production of trace gases and especially aerosol precursors, a study of the oxidation mechanism of alpha- and beta-pinene with hydroxyl radicals must be conducted. The degradation products of both monoterpenes with hydroxyl radicals were identified and quantified in a fast-flow reactor. The products were collected on a liquid nitrogen trap coated with a 2,4-DNPH solution to which two internal standards (benzaldehyde-2,4-DNPH and tolualdehyde-2,4-DNPH) had been added. The collection method was based on the in situ conversion of aldehyde and/or ketone compounds to their 2,4-dinitrophenylhydrazone derivatives. The derivatives were analyzed by HPLC-MS using APCI(-). TIC chromatograms and mass spectral data for the various oxidation products are presented. For alpha-pinene, pinonaldehyde is the most important degradation product, with smaller amounts of acetone, formaldehyde, campholenealdehyde, and acetaldehyde. For beta-pinene, nopinone and formaldehyde are the most abundant products, of almost equal importance, whereas acetone and acetaldehyde are minor compounds. PMID- 11941432 TI - Air monitoring of aldehydes by use of hydrazine reagents with a triazine backbone. AB - Two hydrazine reagents, 4- N, N-dimethylamino-6-(4'-methoxy-1'-naphthyl)-1,3,5 triazine-2-hydrazine (DMNTH) and N-methyl-4- N', N'-dimethylamino-6-(4'-methoxy 1'-naphthyl)-1,3,5-triazine-2-hydrazine (MDMNTH) have been synthesized and used for the determination of aldehydes in air samples. Test tubes with the reagents coated on silica gel were prepared and used for monitoring of carbonyls in air. After elution with acetonitrile the hydrazones formed were separated by reversed phase liquid chromatography. Detection was performed by UV-visible and fluorescence spectroscopy. The results were validated by use of standard atmospheres of the carbonyls and of nitrogen dioxide and ozone, as potential interferents. In comparison with established hydrazine reagents, e.g. 2,4 dinitrophenylhydrazine (DNPH), the results from use of MDMNTH correlate well; lower recoveries were obtained by use of DMNTH. The limits of detection for the new reagents are superior to those for DNPH, because of the possibility of fluorescence detection. PMID- 11941433 TI - Determination of higher carbonyl compounds in used frying fats by HPLC of DNPH derivatives. AB - In current analytical practice, simple methods are required for assessing the quality of a deep-frying fat after some time of use. Therefore, a new procedure was developed for the fast and selective determination of higher carbonyl compounds, i.e., those triglycerides resulting from the oxidation and/or oxidative cleavage of double bonds in unsaturated fatty acids. For analysis, a fat sample is dissolved in a 1-butanol/toluene mixture containing 6-undecanone as an internal standard. Aldehydes and ketones present are allowed to react with 2,4 dinitrophenylhydrazine (DNPH) in acidic solution for at least 1 h at room temperature. The mixture is injected directly onto a reversed phase HPLC column that is eluted with a very steep gradient between methanol and tert-butylmethyl ether (TBME) and is fitted with a UV detector set at 370 nm. In this way, the DNPH derivatives of all higher carbonyl compounds are eluted as one single peak. The result is calculated as milliequivalents of carbonyls per kg of fat (meq kg( 1)). The method takes a minimum of time and reagents and only requires the usual equipment. PMID- 11941434 TI - Comparative studies of the static and dynamic headspace extraction of saturated short chain aldehydes from cellulose-based packaging materials. AB - Aldehydes in cellulose-based materials such as cardboard are derived from lipid degradation. Depending on the production- and storage conditions of the cardboard, the aldehyde content changes. Owing to their sensorial properties, accurate control of their content is obligatory. The cardboard usually exhibits strong and even varying matrix effects and considerable inhomogeneity. The comparability of results of analysis after static and dynamic headspace extraction of short chained saturated aldehydes from cellulose-based matrices was studied. In the case of the static extraction technique, special attention was given to the establishment of the headspace equilibrium, which could be reached by the addition of water as a displacer. For dynamic headspace extraction, the volatiles were purged from the matrix by an inert gas and enriched on an adsorbent trap. In theory, the extraction yield should be 100%. Since there are no certified reference materials for verification of the extraction efficiency available, confirmation was achieved by determining the total amount of analytes in the sample by means of multiple headspace extraction.In comparison to the static operation mode, the major drawbacks of the dynamic technique were found to be based on a more complex parameter string and on limitations to the extractable sample quantities, which may result in enhanced uncertainty of the measurements. Nevertheless, the results of analysis pointed out that both headspace extraction techniques are suitable for the determination of volatile aldehydes from cellulose-based materials. PMID- 11941435 TI - Intermethod comparisons of active sampling procedures and analysis of aldehydes at environmental levels. AB - Within the framework of a European interlaboratory exercise, the Vito facility for the generation of controlled atmospheres was used to test the suitability of four sampling techniques for priority aldehydes namely formaldehyde, acrolein, acetaldehyde, and glutaraldehyde at the ranges of 0.5 to 150 microg x m(-3). The samplers are DNPH-containing impingers, DNPH-impregnated cartridges and filters, and 2-HMP coated XAD-2 tubes. The three first DNPH samplers are to be analyzed by HPLC and the latter by GC-MS for the oxazolidine derivatives. The intermethod comparison comprises two to five sets of experiments depending on the compounds of interest. The aim of the exercise was also to assess the chemical interferences caused by ozone, nitrogen dioxide, and ammonia when using different techniques for sampling and analysis. The active DNPH method (with minor modifications such as shorter sampling time, immediate elution after sampling, and/or eventually wetting of samplers) delivered results within the 30% overall relative uncertainty for formaldehyde, acetaldehyde, and acrolein at the upper microg x m(-3) levels. However, the results suggest that the current DNPH methods for aldehydes do not comply with the 30% minimum performance criteria at the sub microg x m(-3) level. Sampling of aldehydes in the presence of ozone and NO2 interferences by using a "scrubber" cartridge appears to be beneficial to the quality of results. PMID- 11941436 TI - Results from interlaboratory comparisons of aldehyde-2,4 dinitrophenylhydrazone analysis. AB - Within the framework of the SMT "Aldehydes" project (SMT4-CT97-2910) three laboratory comparisons have been organized in which artificial aldehyde hydrazone samples have been distributed among participating laboratories. To these samples potentially interfering compounds have been added to simulate as closely as possible real air or air emission samples.In addition to the "Aldehydes" project partners external EU laboratories have participated. It may, therefore, be assumed that the results of these comparisons reflect the state-of-the-art in the analytical determination of aldehydes in air samples using the 2,4 dinitrophenylhydrazine (DNPH)-method. In this paper several aspects of the organization of these comparisons and the overall results obtained are presented. These demonstrate that results obtained using liquid chromatographic techniques are usually comparable between laboratories within +/-2 times the coefficient of variation (approximately 6-15%), indicating the robustness of the methodology. In addition, laboratory (liquid chromatography) mean results can be made to agree with preparation concentrations at the 95% confidence levels after optimization of separation and integration conditions; these are mainly related to the measurement of formaldehyde in the presence of interferents and the measurement of low concentrations of acrolein and acetone. Some problem areas still remain, however--namely the measurement of glutaraldehyde and the measurement of low levels of acetone. PMID- 11941437 TI - Fiber-optic biosensors--an overview. AB - This article reviews progress and developments during the past five years in the field of optical fiber biosensors. Because of the expense and time constraints associated with modern laboratory analysis, there is a growing need for real time, low-cost technology that can be used industrially, environmentally, and clinically, and to monitor food processing. Miniaturization, integrated systems, and multianalyte determination have become key aspects of sensor development and efforts in this direction will also be discussed, with some pointers to likely directions of future research in the area. The review will provide information about the analytical characteristics and applications of fiber-optic biosensors classified depending on the biorecognition element employed - enzymes, whole cells, antibodies, nucleic acids, and biomimetic polymers. PMID- 11941438 TI - Towards a rehabilitation of the generalized rank annihilation method (GRAM). AB - The trilinear PARAFAC model occupies a central place in multiway analysis, because the components of a data array can often be uniquely resolved. This paper compares the resolution for a large variety of methods, namely the generalized rank annihilation method (GRAM), alternating least squares (ALS), alternating trilinear decomposition (ATLD), alternating coupled vectors resolution (ACOVER), alternating slice-wise diagonalization (ASD), alternating coupled matrices resolution (ACOMAR), self-weighted alternating trilinear decomposition (SWATLD), and pseudo alternating least squares (PALS). The comparison was conducted using Monte Carlo simulations. It was shown that GRAM performs well for moderately and highly overlapped data. These results argue strongly against the previously claimed superiority of the alternatives listed above. PMID- 11941440 TI - Beam-injection flame-furnace atomic-absorption spectrometry (BIFF-AAS) with low pressure sample-jet generation. AB - Flame-furnace AAS (FF-AAS) comprises two individual, recently developed FAAS techniques remarkable for their high power of detection and the small amounts of sample required. One of these techniques is beam-injection flame-furnace AAS (BIFF-AAS), whereby the sample is introduced into a flame-heated atomization cell as a high-speed liquid jet. For the first time this sample-introduction beam has been generated at low pressure by use of a peristaltic pump and a special micro channel smooth jet nozzle developed for this application. Compared with standard FAAS a 7- to 17-fold improvement in the power of detection was obtained for six elements (Ag, Cd, Hg, Pb, Se, and Zn). The relative standard deviation (N=15) was between 0.7% and 3.4%. Sample volumes between 10 microL and 1 mL have been investigated. The applicability of the peristaltic pump for beam generation enables an optimum sampling mode for a variety of analytical tasks (continuous sample uptake, manual or automated, or different flow-injection modes). The method can also be regarded as a simple, effective interface between FIA techniques and flame AAS. PMID- 11941439 TI - Polarization immunoassays using reactive ruthenium metal-ligand complexes as luminescent labels. AB - The first competitive fluorescence polarization immunoassay using ruthenium metal ligand complexes (5-MC and 55-DC) as labels is described. These were newly synthesized and characterized in terms of their spectra, their covalent linkage to proteins, and their use in both homogeneous and competitive immunoassays. Linkage to proteins was achieved by the N-hydroxysuccinimide ester method, which was demonstrated for the systems HSA-anti-HSA and myoglobin-anti-myoglobin. The values of the fundamental polarization are 0.18 for 5-MC and 0.33 for 55-DC. Polarization immunoassays with labeled HSA and myoglobin were performed in the homogeneous format and resulted in an increase of the fluorescence polarization of up to 100%. In the competitive assay, a decrease in polarization of >90% was detectable. When the competitive HSA immunoassay was validated against an independent test both methods gave almost the same results. PMID- 11941441 TI - Optimisation of sample pre-treatment in the HG-AAS selenium analysis. AB - A fast and accurate method of pre-treatment for biological materials in the hydride generation-atomic absorption spectroscopy (HG-AAS) selenium determination is described. All procedures of sample pre-treatment were performed in closed vessels in a microwave oven. After sample decomposition using a HNO3/H2O2 mixture, the pre-reduction and a 'denitrification' step (reduction of interfering nitrogen intermediates) with amidosulfuric acid were combined. Hence, analysis time and the risk of element losses or the emergence of possible contamination are minimised. PMID- 11941442 TI - Effect of the porosity of PS-DVB-copolymers on ion chromatographic behavior in inverse size-exclusion and ion chromatography. AB - Small and highly pressure-stable PS-DVB copolymers of different porosity had been prepared by a two-step swelling procedure which enabled variation of diluent composition, an important characteristic affecting the porosity. The polymers were characterized by inverse size-exclusion chromatography and scanning electron microscopy. Subsequent chloromethylation and amination resulted in anion exchangers suitable for ion chromatography. The pore volume and the pore-size distribution is substantially affected by the fraction of the solvens component in the diluent. It was apparent from scanning electron microscopy that surface structure and the size of the polymer particles was not affected by diluent composition. The functionalization process led to a decrease in pore volume. The pore-size distribution remained unchanged during functionalization, which can be explained in terms of partial closing of all pore sizes. The chromatographic efficiency of the functionalized polymers in ion chromatography was highly dependent on diluent composition and the extent of functionalization was determined by the total pore volume. The composition of the diluent is an excellent tool for optimization of polymers used for the synthesis of surface functionalized anion exchangers. PMID- 11941443 TI - Sol-gel derived carbon ceramic electrode containing methylene blue-intercalated alpha-zirconium phosphate micro particles. AB - Methylene blue-intercalated alpha-zirconium phosphate (MBZrP) micro particles in deionized water were deposited onto the surface of graphite powder to prepare graphite powder-supported MBZrP, which was subsequently dispersed into methyltrimethoxysilane-derived gels to yield a conductive composite. The composite was used as electrode material to fabricate a surface-renewable, rigid, leak-free carbon ceramic composite electrode, bulk-modified with methylene blue (MB). In the configuration, alpha-zirconium phosphate was employed as a solid host for MB, which acted as a catalyst. Graphite powder ensured conductivity by percolation, the silicate provided a rigid porous backbone and the methyl groups endowed hydrophobicity and thus limited the wetting section of the modified electrode. Peak currents of the MBZrP-modified electrode were surface-confined at low scan rates but diffusion-controlled at high scan rates. Square-wave voltammetric study revealed that MBZrP immobilized in carbon ceramic matrix presented a two-electron, three-proton redox process in acidic aqueous solution with pH ranged from 0.44 to 2.94. In addition, the chemically modified electrode showed an electrocatalytic activity toward nitrite reduction at +0.15 V ( vs. Ag/AgCl) in acidic aqueous solution (pH=0.44). The linear range and detection limit are 1x10(-6)-4x10(-3) mol x L(-1) and 1.5x10(-7) mol x L(-1), respectively. PMID- 11941444 TI - Coated-wire copper(II)-selective electrode based on phenylglyoxal-alpha-monoxime ionophore. AB - A copper(II) ion-selective electrode based on a recently synthesized 2-quinolyl-2 phenylglyoxal-2-oxime (phenylglyoxal-alpha-monoxime) has been developed. The PVC based membrane containing phenylglyoxal-alpha-monoxime, dibutyl phthalate as plasticizer, and sodium tetraphenylborate as anion excluder and membrane modifier, was directly coated on the surface of a platinum-wire electrode. The response of the electrode was linear with a near-Nernstian slope of 28.2 mV decade(-1) within the Cu2+ ion concentration range 1x10(-6)-1x10(-1) mol x L(-1). The response time for the proposed electrode to achieve a 95% steady potential for Cu2+ concentrations ranging from 1x10(-1) to 1x10(-6) mol x L(-1) is between 10 and 50 s, and the electrode is suitable for use within the pH range of 3 to 6.5. The electrode has a detection limit of 5x10(-7) mol x L(-1) Cu2+ and its selectivity relative to several alkali, alkaline earth, transition, and heavy metal ions was good. The coated-wire electrode could be used for at least two months without a considerable alteration of its potential. Applications of the electrode for determination of copper in milk powder samples and as an indicator electrode for potentiometric titration of Cu2+ ion using EDTA are reported. PMID- 11941445 TI - Slurry sampling for the determination of arsenic, cadmium, and lead in mainstream cigarette smoke condensate by graphite furnace-atomic absorption spectrometry and inductively coupled plasma-mass spectrometry. AB - The slurry sampling technique has been applied for the determination of As, Cd, and Pb in mainstream cigarette smoke condensate (MS CSC) by graphite furnace atomic absorption spectrometry (GF-AAS) and inductively coupled plasma-mass spectrometry (ICP-MS). The MS CSC of the 1R4F Reference Cigarette was collected by electrostatic precipitation and was subsequently prepared as two slurry samples with and without the dispersing agent Triton X-100. Comparison of results determined by ICP-MS analyses of the 1R4F MS CSC slurry samples with those from the conventional microwave digestion method revealed good agreement. The precision of Triton X-100 slurry sampling and of microwave-assisted digestion was better than 10% RSD, and both were superior to slurry sampling without use of Triton X-100. The accuracy of the analytical results for the Triton X-100 slurry sample was further verified by graphite furnace-atomic absorption spectrometry (GF-AAS). For GF-AAS, the method limits of detection are 1.6, 0.04, and 0.5 microg x L(-1) for As, Cd, and Pb, respectively. For ICP-MS, the method limits of detection are 0.06, 0.01, and 0.38 microg x L(-1) for As, Cd, and Pb, respectively. The MS CSC of the 1R4F Reference Cigarette was collected in accordance with the Federal Trade Commission (FTC) smoking regime (35 mL puff volume of 2-s puff duration at an interval of 60 s) and the concentrations of As, Cd and Pb were 6.0+/-0.5, 69.3+/-2.8, and 42.0+/-2.1 ng/cigarette, respectively. PMID- 11941446 TI - Determination of pesticide residues in red wines with microporous membrane liquid liquid extraction and gas chromatography. AB - A sample pretreatment method based on microporous membrane liquid-liquid extraction (MMLLE) was developed for the subsequent gas chromatographic determination of pesticides in wine. MMLLE provided efficient and selective extraction with enrichment factors in the range 3-13. The gas chromatographic separation was carried out using on-column injection and flame ionization detection. The method was linear, repeatable and sensitive. The limits of quantification were better than 0.006 mg/L for all the analytes except for iprodione (0.37 mg/L). The method was applied to the determination of pesticides in several red wines of different origin. PMID- 11941447 TI - A BOD biosensor based on a microorganism immobilized on an Al2O3 sol-gel matrix. AB - A novel BOD biosensor has been fabricated from a microbial membrane on an oxygen electrode; a porous inorganic Al2O3 sol-gel matrix was used to immobilize the yeast. Use of the sensor is convenient and rapid compared with the official BOD5 method. The gelation time required to form the microorganism membrane was less than 15 min. There was linear relationship between the response (sensor current) and BOD values ranging from 10-50 mg x L(-1). The lifetime of the BOD biosensor was more than 30 days at ambient temperature. PMID- 11941448 TI - Simultaneous determination of gold(III), palladium(II), and platinum(IV) with N phenyl- N'-(sodium p-aminobenzenesulfonate)thiourea. AB - The preparation and characteristics of a new water-soluble reagent, N-phenyl- N' (sodium p-aminobenzenesulfonate)thiourea (PPT) are described. In the presence of cetyltrimethylammonium bromide (CTMAB) PPT reacts with Au(III), Pd(II), and Pt(IV) to form colored complexes with absorption maxima at 317 nm, 306.1 nm, 778.4 nm, respectively. Optimum conditions for color development were studied. The reagent was used for the simultaneous determination of Au(III), Pd(II), and Pt(IV); Amberlyst A-26 macropore anion-exchange resin was used as a means of rapid separation. The method was applied to the determination of Au(III), Pd(II), and Pt(IV) in catalyst materials and anode mud with satisfactory results. PMID- 11941449 TI - Highly selective and sensitive reaction of cyanide with 2,2-dihydroxy-1,3 indanedione. AB - A novel reaction of cyanide with 2,2-dihydroxy-1,3-indanedione in the presence of sodium carbonate is described. It is highly selective and sensitive, and suitable for the determination of hydrogen cyanide in the environment and free cyanide ions in water, blood, urine, serum, etc. As little as 1.25x10(-7) mol x L(-1) CN( ) (3.25x10(-9) g x mL(-1) cyanide) can be determined by use of this reaction. The color system obeys Beer's law in the range 10 ng x mL(-1) to 1.0 microg x mL(-1) at 510 nm. The molar absorptivity was 8.0x10(4) L x mol(-1) x cm(-1) for a solution of concentration 0.2 microg x mL(-1). All other important analytical properties of the reaction have been studied. It is proposed that the purple color produced under these reaction conditions is that of 2-cyano-1,2,3 trihydroxy-2 H indene. PMID- 11941450 TI - Monoclonal antibody to a cancer-specific and drug-responsive hydroquinone (NADH) oxidase from the sera of cancer patients. AB - Monoclonal antibodies were generated in mice to a 34-kDa circulating form of a drug-responsive hydroquinone (NADH) oxidase with a protein disulfide-thiol interchange activity specific to the surface of cancer cells and the sera of cancer patients. Screening used Western blots with purified 34-kDa tNOX from HeLa cells and the sera of cancer patients. Epitopes were sought that inhibited the drug-responsive oxidation of NADH with the sera of cancer patients, but which had no effect on NADH oxidation with the sera of healthy volunteers. Two such antisera were generated. One, designated monoclonal antibody (mAb) 12.1, was characterized extensively. The NADH oxidase activity inhibited by mAb 12.1 also was inhibited by the quinone site inhibitor capsaicin (8-methyl- N-vanillyl-6 noneamide). The inhibition was competitive for the drug-responsive protein disulfide-thiol interchange activity assayed either by restoration of activity to scrambled RNase or by cleavage of a dithiodipyridine substrate, and was uncompetitive for NADH oxidation. Both the mAb 12.1 and the postimmune antisera immunoprecipitated drug-responsive NOX activity and identified the same 34-kDa tNOX protein in the sera of cancer patients that was absent from sera of healthy volunteers, and was utilized as immunogen. Preimmune sera from the same mouse as the postimmune antisera was without effect. Both mouse ascites containing mAb 12.1 and postimmune sera (but not preimmune sera) slowed the growth of human cancer cell lines in culture, but did not affect the growth of non-cancerous cell lines. Immunocytochemical and histochemical findings showed that mAb 12.1 reacted with the surface membranes of human carcinoma cells and tissues. PMID- 11941451 TI - Beta1 integrin triggering affects leukemic cell line sensitivity to natural killer cells. AB - The role of beta1 (CD29) integrins in natural killer (NK) cell-target cell conjugation and cytotoxicity has not been clearly established. Ligation of beta1 integrins in NK cells can modulate the lytic capacity in both a positive and a negative manner; however, the contribution of the beta1 integrins present on target cells remains to be evaluated. Here, we analyzed the effect of beta1 integrins expressed by potential tumor target cells on conjugation and cytotoxicity. Using normalized flow cytometry binding assays, we demonstrated that the pretreatment of MOLT-4, K562, U-937 and HL-60 human leukemia target cell lines with selected anti-beta1 monoclonal antibodies (mAb) increased conjugation to human NK cell line NKL as well as to purified NK cells. Only mAb recognizing residues 207-218 of the beta1 subunit and functionally involved in the induction of homotypic adhesion (functional epitope A1) increased conjugation of all the target cells. Moreover, mAb to adhesion molecules different from beta1 but also inducers of homotypic adhesion of the target cells, i.e. CD43 and CD50 (ICAM-3), failed to increase conjugation to NKL cells. Cytotoxicity assays demonstrated that lysis of NK-sensitive target cells (MOLT-4) also increased after pretreatment with anti-beta1 epitope A1 mAb. Importantly, pretreatment of NK resistant target cells (U-937 and HL-60) with anti-beta1 mAb was not able to outweigh the cytotoxic inhibitory mechanisms controlled by HLA class I molecules. However, simultaneous masking of HLA class I molecules with mAb and pretreatment with anti-beta1 mAb rendered NK-resistant cells susceptible to lysis, as predicted by the missing self hypothesis. Triggering of tumor target cells through beta1 integrins may thus play a role in conjugation to NK cells as well as in co-stimulation of cell-mediated cytotoxicity. PMID- 11941452 TI - Mature dendritic cells are protected from Fas/CD95-mediated apoptosis by upregulation of Bcl-X(L). AB - The clinical use of dendritic cells (DC) as tumor vaccines is very much dependent on their survival potential. Members of the tumor necrosis factor (TNF) receptor superfamily and their ligands are involved in the regulation of cell death. Fas (CD95) is a representative protein that promotes apoptosis. The Bcl-2 family of proteins functions as an integrator of diverse pro- and anti-apoptotic signals. It has been found that DC maturation facilitates their survival, and thus has an anti-apoptotic function. However, little is known about the underlying mechanisms. We investigated the effects of TNF-alpha and lipopolysaccharide (LPS) on the expression of apoptotic molecules during differentiation and maturation of DC under serum-free conditions, and correlated this to the sensitivity to apoptosis by the Fas-mediated pathway. Indeed, DC activation effectively inhibited DC apoptosis, which was predominantly accompanied by the upregulation of Bcl-X(L) and to a lesser extent Bcl-2, while Bax and FLICE inhibitory protein (FLIP) remained unchanged. In contrast, in the presence of serum FLIP was also upregulated. We conclude that under serum-free conditions, Bcl-X(L) rather than FLIP plays the main role in protection against DC apoptosis. PMID- 11941453 TI - Inducing specific reactivity against B cells in mice by immunizing with an Fc fusion protein containing self-Igbeta. AB - A recombinant chimeric fusion protein, muIgbeta-hugamma4.Fc, composed of the extracellular domain of mouse Igbeta (CD79b) and the CH2-CH3 domains of human IgGgamma4.Fc (hugamma4.Fc), linked via an immunologically inert flexible peptide, was prepared. The fusion protein was evaluated for its ability to induce specific auto-reactive immune response against Igbeta and to modulate B cell activity in Balb/c mice. Upon immunization with muIgbeta-hugamma4.Fc, mice developed immunoglobulin (IgG) against self-Igbeta, which could bind to the cells of a mouse B cell line expressing Igbeta on the cell surface. The proportion of B cells in mononuclear cells in the peripheral blood (PBMC) of treated mice decreased as compared to that of mice immunized with hugamma4.Fc without the Igbeta component. Furthermore, mice immunized against muIgbeta-hugamma4.Fc displayed a reduced antibody response against an irrelevant antigen. The implications of employing the present approach in developing a therapeutic strategy for regulating B cell activity has been discussed. PMID- 11941454 TI - Folate targeting of haptens to cancer cell surfaces mediates immunotherapy of syngeneic murine tumors. AB - A variety of human cancers overexpress a cell surface receptor with high affinity for the vitamin, folic acid ( K(d) approximately 10(-10)M). Covalent attachment of therapeutic agents to folic acid has been shown to allow efficient targeting of the folate-drug conjugates to folate receptor-expressing cancer cells, with little or no uptake by normal tissues except the kidneys. We report here the use of folate's ability to deliver attached molecules specifically to cancer cells to convert poorly immunogenic tumors into highly immunogenic tissue targets. By linking folic acid to a model hapten, we have been able to decorate folate receptor-expressing cancer cell surfaces with >10(6) haptens/cell in vivo. Following marking of such cells with haptens, the cells are observed to become opsonized with autologous anti-hapten antibodies, which is presumed to mediate cell removal via antibody-dependent cellular cytotoxicity (ADCC). Supplemental administration of low levels of ADCC-activating cytokines [e.g. interleukin-2 (IL 2) and interferon-alpha (IFN-alpha)] has been shown to synergize with the folate targeted immunotherapy. Thus, using M109 syngeneic lung cancer cells injected intraperitoneally into Balb/c mice that were previously immunized against fluorescein, a significant extension of life span is observed following treatment with folate-fluorescein conjugates, and complete cures are observed upon supplementation with moderate levels of IL-2 and IFN-alpha. Because control tumor bearing mice treated with the same cytokines but with non-targeted fluorescein show no extension of life span, we conclude that tumor-specific opsonization is an essential step in this immunotherapy. Finally, because the anti-fluorescein antibodies are unable to access the folate receptors on the apical membranes of the kidney proximal tubules, no kidney or other normal tissue cytotoxicity is observed. These data suggest that retargeting of haptens to folate receptor expressing cancers might constitute a method for mobilizing the immune system specifically against poorly immunogenic tumors. PMID- 11941455 TI - Immunization with heat shock protein 70 from methylcholanthrene-induced sarcomas induces tumor protection correlating with in vitro T cell responses. AB - The cytosolic members of the heat shock protein 70 (hsp70) family have in recent years been shown to elicit protective immunity mediated via binding to antigenic peptides in tumor and viral animal models. In this study we have used the methylcholanthrene (MC)-induced sarcomas MC57S and MC57X, previously shown to express individually distinct MHC-I associated peptides recognized by tumor necrosis factor-alpha (TNF-alpha) producing CD8(+) T cells. With hsp70 purified from tumor or liver tissue, we were able to confirm that tumor-derived hsp70 elicited in vivo protection against a challenge with the same tumor as that used for hsp70 isolation. We also observed that immunizing with hsp70 isolated from tumor tissue resulted in a significantly better protection than immunizing with hsp70 isolated from the liver tissue of healthy mice. In two out of three experiments however, immunization with liver-derived hsp70 as well as tumor derived hsp70 resulted in a significantly delayed tumor outgrowth as compared to saline-injected controls. In vitro, T cell lines from mice immunized with tumor derived hsp70 could recognize tumor cells from the same MC57 tumor as that used for the hsp70 purification, resulting in TNF-alpha production. In sera from hsp70 immunized mice we observed undetectable or only very low levels of anti-hsp70 antibodies, suggesting that it is possible to repeatedly immunize the mice with no significant interference from neutralizing antibodies. We therefore conclude that hsp70 immunization can lead to protection against the tumor it was purified from, with a low risk of eliciting neutralizing antibodies that could affect further immunizations. PMID- 11941456 TI - Anti-renal cell carcinoma chimeric antibody G250: cytokine enhancement of in vitro antibody-dependent cellular cytotoxicity. AB - The chimeric monoclonal antibody cG250 targets the G250 antigen, a transmembrane protein which is expressed on renal carcinoma cells and is identical to the MN/CAIX antigen. In vitro studies have previously demonstrated that cG250 induces antibody-dependent cellular cytotoxicity (ADCC) of G250-positive targets. In order to investigate the upregulation of ADCC mediated by cG250, ADCC was examined using effector cells cultured in the presence or absence of the cytokines interferon-gamma (IFN-gamma), interferon-alpha isoforms IFN-alpha (2a) and IFN-alpha (2b) and interleukin-2 (IL-2), and the time course of effects over a 7-day period was determined. Renal cell carcinoma lines expressing high (SK-RC 52) and low (SK-RC-09) G250 antigen levels were used as target cells, and freshly isolated peripheral blood mononuclear cells (PBMC) from a healthy donor were used as the effector cells. PBMC were incubated with the respective cytokine at a range of concentrations or with a media alone control for a period of 7 days. The ADCC activity mediated by cG250 or control isotype matched huA33 with the different PBMC treatment groups was assessed in triplicate daily. Corresponding lymphokine activated killing (LAK) activity was measured concurrently for each treatment group. Chimeric G250 specifically recognised G250 antigen on high and low expressing cell lines SK-RC-52 and SK-RC-09, and mediated specific in vitro ADCC of both lines. In the absence of cytokine stimulation, the specific ADCC of cG250 declined rapidly within three days. IL-2 strongly enhanced and maintained cG250-mediated ADCC activity and K562 cytotoxicity when applied to PBMC in culture for seven days. IFN-gamma also enhanced the ADCC of cG250 throughout the study period, but was not as effective as the IL-2 treatment, and the SK-RC-09 line displayed lower specific cytotoxicity than the SK-RC-52 cell line. In contrast, IFN-alpha 2a and 2b increased cG250-mediated ADCC and K562 cytotoxicity for only three days of the study period. The potent and sustained immune effector activity observed with cG250 and cytokines in this in vitro study suggests that the combination immunotherapy of cG250 with cytokines such as IL-2 shows promise in the treatment of renal cell carcinoma (RCC). PMID- 11941457 TI - Evidence that hydroxyl radicals mediate auxin-induced extension growth. AB - Reactive oxygen intermediates, i.e. the superoxide radical (O*-)(2), hydrogen peroxide (H2O2) and the hydroxyl radical (*OH), are generally regarded as harmful products of oxygenic metabolism causing cell damage in plants, animals and microorganisms. However, oxygen radical chemistry may also play a useful role in polymer breakdown leading to wall loosening during extension growth of plant cells controlled by the phytohormone auxin. Backbone cleavage of cell wall polysaccharides can be accomplished in vitro by (*OH) produced from H2O2 in a Fenton reaction or in a reaction catalyzed by peroxidase supplied with O2 and NADH. Here, we show that coleoptile growth of maize seedlings is accompanied by the release of reactive oxygen intermediates in the cell wall. Auxin promotes release of (O*-)(2) and subsequent generation of (*OH)when inducing elongation growth. Experimental generation of (*OH) in the wall causes an increase in wall extensibility in vitro and replaces auxin in inducing growth. Auxin-induced growth can be inhibited by scavengers of (O*-)(2), H2O2 or (*OH), or inhibitors interfering with the formation of these molecules in the cell wall. These results provide the experimental background for a novel hypothesis on the mechanism of plant cell growth in which (*OH), produced from (O*-)(2) and H2O2 by cell wall peroxidase, acts as a wall-loosening agent. PMID- 11941458 TI - Somatic embryogenesis from Arabidopsis shoot apical meristem mutants. AB - Zygotic embryos of three Arabidopsis thaliana (L.) Heynh. mutants lacking an embryonic shoot apical meristem (SAM), shoot meristemless (stm), wuschel (wus) and zwille/pinhead (zll/pnh) were used as explants to establish embryogenic cell cultures. Somatic embryos of all three mutants showed the same mutant phenotypes as their zygotic equivalents. These results provide genetic evidence that the developmental program of somatic and zygotic embryos is indistinguishable. They also suggest that a functional SAM is not required for somatic embryogenic cell formation in Arabidopsis. PMID- 11941459 TI - A dual effect on protein synthesis and degradation modulates the tubulin level in rice cells treated with oryzalin. AB - The effect of the anti-microtubular drug oryzalin on growth and morphology of cultured rice (Oryza sativa L., cv. Roncarolo) cells was evaluated with specific reference to mechanisms that control intracellular tubulin levels. The addition of oryzalin caused a great reduction in the level of both alpha- and beta-tubulin polypeptides, as detected by Western blot analysis. However, no appreciable decrease was observed in the population of total or isotype-specific alpha- and beta-tubulin mRNAs. Only within the first 24 h of the oryzalin treatment, when the level of both alpha- and beta-tubulin polypeptides was still undiminished, was a consistent reduction in the amount of total beta-tubulin mRNA observed. Pulse-chase experiments performed on rice cells grown in the presence of 1 microM oryzalin revealed the presence of two distinct mechanisms that negatively control alpha- and beta-tubulin polypeptide levels. (i) There was an immediate effect on protein synthesis, which resulted in a reduction in the level of newly synthesized tubulin. (ii) There was a delayed response characterized by a substantial degradation of both alpha- and beta-tubulin monomers; this degradation occurred after 24 h of herbicide treatment. The possible involvement of Ca2+ in the degradation of the unincorporated tubulin monomers is also documented and discussed. PMID- 11941460 TI - Chitinase production in pine callus (Pinus sylvestris L.): a defense reaction against endophytes? AB - In shoot tip-derived tissue cultures of Scots pine (Pinus sylvestris L.), browning and subsequent degeneration of the culture is accompanied by lipid peroxidation and lignification of cells, which are characteristic features of a plant defense reaction. Since chitinases are enzymes acting primarily in plant defense, their expression was studied in pine callus in order to elucidate the defense reaction. Chitinases were present diversely in tissue cultures originating from shoot tips and embryos of P. sylvestris, in contrast to Pinus nigra embryogenic callus, where production of chitinases or browning was not detected. Because endophytic microbes had earlier been detected in buds of Scots pine, their subsequent presence in the tissue cultures was considered a potential cause of the defense reaction. Therefore, the presence of endophytes in the tissue cultures was examined by in situ hybridization. Endophytes were found to colonize heavily in 45% of the tissue cultures of P. sylvestris and to form biofilms, while the P. nigra callus was not found to contain any microbes. The endophytes seemed to propagate uncontrollably once a tissue culture of P. sylvestris was initiated. Regardless of the high level of chitinase production in the callus, the control of the endophytes presumably becomes inadequate during the tissue culture of P. sylvestris. PMID- 11941461 TI - Biochemical, molecular and structural analysis of multiple thaumatin-like proteins from the elderberry tree (Sambucus nigra L.). AB - Thaumatin-like proteins (TLPs) were isolated and characterized from fruits and leaves of elderberry (Sambucus nigra) and their corresponding genes cloned. In addition, the developmental regulation and induction of the different TLPs was followed in some detail. Ripening berries accumulated a fruit-specific TLP during the final stages of maturation. This fruit-specific TLP had no antifungal activity and was devoid of beta-glucanase activity. Leaves constitutively expressed a TLP that closely resembled the fruit-specific homologue. Treatment with jasmonate methyl ester induced two additional TLPs in leaves but did not induce or enhance the expression of TLPs in immature berries. In contrast to jasmonate methyl ester, both ethephon and garlic extract induced the expression of a TLP in unripe berries that normally do not express any TLP. Sequence analysis and molecular modeling indicated that all elderberry thaumatin-like proteins share a high sequence similarity with group-5 pathogenesis-related proteins. However, the proteins encoded by the different sequences differed from each other in isoelectric point and the distribution of the charges on the surface of the molecule. PMID- 11941462 TI - Photosynthetic control of the plasma membrane H+-ATPase in Vallisneria leaves. I. Regulation of activity during light-induced membrane hyperpolarization. AB - In mesophyll cells of the aquatic angiosperm Vallisneria gigantea Graebner, red, blue, or blue plus far-red light induced a typical membrane hyperpolarization, whereas far-red light alone had little effect. Both N,N' dicyclohexylcarbodiimide, a potent inhibitor of H+-ATPase, and carbonylcyanide m chlorophenylhydrazone, an uncoupler, produced a considerable membrane depolarization in the dark-adapted cells and a complete suppression of the light induced hyperpolarization. Although 3-(3',4'-dichlorophenyl)-1,1-dimethylurea (DCMU), an inhibitor of photosynthetic electron transport, did not affect the membrane potential in darkness, it completely inhibited the light-induced membrane hyperpolarization. In vivo illumination of the leaves with red light caused a substantial decrease in the Km for ATP, not only of the vanadate sensitive ATP-hydrolyzing activity in leaf homogenate, but also of the ATP dependent H+-transporting activity in plasma membrane (PM) vesicles isolated from the leaves by aqueous polymer two-phase partitioning methods. The effects of red light were negated by the presence of DCMU during illumination. In vivo illumination with far-red light had no effect on the Km for ATP of H+ transporting activity. These results strongly suggest that an electrogenic component in the membrane potential of the mesophyll cell is generated by the PM H+-ATPase, and that photosynthesis-dependent modulation of the enzymatic activity of the PM H+-ATPase is involved in the light-induced membrane hyperpolarization. PMID- 11941463 TI - Photosynthetic control of the plasma membrane H+-ATPase in Vallisneria leaves. II. Presence of putative isogenes and a protein equipped with a C-terminal autoinhibitory domain. AB - In vitro treatment with trypsin of plasma membrane (PM) vesicles isolated from the leaves of Vallisneria gigantea Graebner, an aquatic monocot, produced a marked decrease in the Km for ATP and an increase in the Vmax of H+-transporting activity. Concomitantly, the removal of 8 kDa of the C-terminal domain from the 94-kDa PM H+-ATPase was confirmed by immunoblotting using different kinds of polyclonal antibody. Three partial clones of putative PM H+-ATPase genes (Vga1, 2, and 3) were isolated from leaves by reverse transcription polymerase chain reaction. Northern blotting analysis revealed that the expression level of Vga3 was high and that of the other two genes was much lower. The H+-transporting activity of PM vesicles was substantially suppressed in the presence of inorganic phosphate (Pi), which has been supposed to be a noncompetitive inhibitor of the PM H+-ATPase, coincident with an increase in the Km for ATP and a decrease in the Vmax. After treatment of the isolated PM vesicles with trypsin, the inhibitory effect of Pi was no longer evident. This result indicates that Pi inhibited the activity through the C-terminal autoinhibitory domain of the PM H+-ATPase. Furthermore, Pi increased the Km for ATP of the H+-transporting activity in the PM vesicles isolated from both dark-adapted and red-light-irradiated leaves. The results suggest that regulation of the Km for ATP through the operation of photosynthesis is independent of regulation through the cytoplasmic level of Pi. PMID- 11941464 TI - Photorespiration-dependent increases in phospho enolpyruvate carboxylase, isocitrate dehydrogenase and glutamate dehydrogenase in transformed tobacco plants deficient in ferredoxin-dependent glutamine-alpha-ketoglutarate aminotransferase. AB - The metabolic cross-talk associated with re-assimilation of photorespiratory NH4+ was analysed in transformed tobacco (Nicotiana tabacum L.) plants with low activities of ferredoxin-dependent glutamine-alpha-ketoglutarate aminotransferase (Fd-GOGAT; EC 1.4.7.1). Amounts of ribulose-1,5-bisphosphate carboxylase oxygenase (Rubisco; EC 4.1.1.39) protein and Rubisco transcripts were similar in all lines whether photorespiration rates were low (4,000 microl l(-1) CO2) or high (air). Leaf sucrose, hexose and starch contents were similar in all lines. In contrast, there was evidence that anaplerotic carbon flow was stimulated in the transformed lines with less than 60% Fd-GOGAT, since phospho enolpyruvate carboxylase (PEPc) activity and (PEPc) protein were increased. A strong positive correlation between leaf PEPc activity and glutamine accumulation was observed, suggesting that the increase in PEPc was related to the accumulation of glutamine. A modest stimulation of total NADP-isocitrate dehydrogenase (ICDH; EC 1.1.1.42) activity was also observed in the transformed lines with less than 60% Fd-GOGAT. This was accompanied by increases in both the cytosolic ICDH and mitochondrial NAD-isocitrate dehydrogenases (IDH; EC 1.1.1.41). IDH protein was also increased in the transformed plants with low Fd-GOGAT, suggesting that both IDH and ICDH are involved in the production of carbon skeletons (and ultimately alpha-ketoglutarate) necessary for the re-assimilation of NH4+. In contrast, PEPc, ICDH and IDH transcripts were similar in all lines. The aminating (but not the de-aminating) activity of NAD(H)-glutamate dehydrogenase (NAD(H)-GDH; EC 1.4.1.2) was greatly increased in plants with less than 60% of Fd-GOGAT after transfer to air. The data confirm that NH4+ or glutamine are involved in signalling, leading to modified gene expression and enzyme activity required for enhanced production of the C skeletons, to accommodate increases in the assimilation of photorespiratory NH4+. In addition, we provide the first demonstration of a compensatory role for NAD(H)-GDH in stabilising the leaf glutamic acid pool when Fd-GOGAT becomes limiting. PMID- 11941465 TI - Sugar exudation by roots of kallar grass [Leptochloa fusca (L.) Kunth] is strongly affected by the nitrogen source. AB - Exudation of sugars (glucose, fructose and sucrose) and that of cations and anions from intact roots of kallar grass [Leptochloa fusca (L.) Kunth] grown hydroponically with ammonium or nitrate (3 mM) as N source was investigated. In different experiments, plants grown on ammonium had slightly higher sugar contents than nitrate-grown plants, but their total sugar exudation during a 2-h period was up to 79-fold higher than under nitrate nutrition. Relative root exudation of inorganic anions and cations and that of amino acids (as a percentage of the internal contents exuded per time) was either similar or slightly higher from ammonium-grown than from nitrate-grown plants. Analysis of root architectural parameters revealed that ammonium-grown plants had a higher number of root tips/side roots per gram root fresh weight than nitrate-grown plants, whereas other root parameters, viz. length, diameter, volume and surface area were similar under the two N sources. A majority of the fine roots having diameter up to 0.4 mm represented up to 86% of the total root length, 64% of the total root surface area, and 35% of the total root volume; the root length and surface area per root system of that major root population were similar in ammonium- and nitrate-grown plants. Apparently, root architecture was not responsible for the different exudation rates. Within 12-24 h after shifting ammonium-grown plants to nitrate nutrition, root sugar levels and visible root architecture remained unchanged, yet the sugar exudation rate was reduced 30 fold. Short-term uptake of [14C]glucose (10 microM) from the rooting medium was similar for ammonium- and nitrate-grown plants. Thus, the very different sugar exudation rates were neither related to internal root sugar concentration, nor to the different root architecture, nor to differential resorption of sugars by ammonium- versus nitrate-grown plants. Increased external Ca2+ did not alter sugar exudation, and decreased external pH (4.5) only slightly increased sugar exudation from roots of nitrate-grown plants kept at pH 6.5. It is suggested that the much higher sugar exudation in response to ammonium may facilitate the ecologically and economically important association of diazotrophs with kallar grass roots. PMID- 11941466 TI - Jasmonate and salicylate induce the expression of pathogenesis-related-protein genes and increase resistance to chilling injury in tomato fruit. AB - Treatment of tomato (Lycopersicon esculentum L. cv. Beefstake) fruit with low concentrations of (0.01 mM) methyl jasmonate (MeJA) or methyl salicylate (MeSA) substantially enhanced their resistance to chilling temperature and decreased the incidence of decay during low-temperature storage. While studying the expression of pathogenesis-related (PR) protein genes, different accumulation patterns of PR protein mRNAs in tomato fruit were observed. MeJA substantially increased the accumulation of PR-2b transcripts encoding intracellular beta-1,3-glucanase and enhanced the mRNA levels of PR-2a and PR-3b encoding extracellular beta-1,3 glucanase and intracellular chitinase, respectively. MeSA substantially increased accumulation of PR-2b and PR-3a mRNAs and slightly increased PR-3b mRNA accumulation. Chilling temperature did not appreciably enhance the accumulation of PR-protein mRNAs in untreated fruit. However, the accumulation of PR-3b mRNAs in MeSA-treated fruit was enhanced following low-temperature storage. Transcript abundance of catalase genes also was investigated in different pretreated tomatoes. The accumulation of cat1 mRNA was increased substantially by MeJA, while it was reduced by MeSA treatment. These results suggest that the pre treatment of tomato fruit with MeSA or MeJA induces the synthesis of some stress proteins, such as PR proteins, which leads to increased chilling tolerance and resistance to pathogens, thereby decreasing the incidence of decay. PMID- 11941467 TI - Over-expression of cinnamate 4-hydroxylase leads to increased accumulation of acetosyringone in elicited tobacco cell-suspension cultures. AB - Cell-suspension cultures were produced from transgenic tobacco (Nicotiana tabacum L.) plants harboring a constitutively expressed alfalfa cinnamate 4-hydroxylase (C4H) transgene. Increased levels of C4H enzyme activity in the transgenic cultures were observed only following exposure of the cells to yeast elicitor, although alfalfa C4H transcripts were expressed at a high level from the cauliflower mosaic virus 35S promoter in the absence of elicitation. Increased expression of C4H in elicited cell-suspension cultures had no appreciable effect on the HPLC profiles of soluble phenolic compounds. However, levels of one compound, subsequently identified as 3,5-dimethoxy-4-hydroxy acetophenone (acetosyringone), were strongly elevated in the wall-bound phenolic fraction. The results are discussed in relation to the correlation between C4H activity and the synthesis of 3,5-dimethylated hydroxycinnamic acid derivatives in tobacco. PMID- 11941468 TI - Evidence for the involvement of tetrahydrofolate in the demethylation of nicotine by Nicotiana plumbaginifolia cell-suspension cultures. AB - The conversion of nicotine to nornicotine by Nicotiana plumbaginifolia Viv. cells was investigated by analysing the redistribution of label during feeding experiments with (R,S)-[2H- methyl]nicotine, (R,S)-[13C- methyl]nicotine and (R,S)-[14C- methyl]nicotine, and the results show that the N-methyl group of nicotine can be recycled into primary metabolism. Nuclear magnetic resonance (NMR) analysis of ethanolic extracts of cells grown in the presence of (R,S)-[13C methyl]nicotine, using 1H-13C correlation spectroscopy (HMQC, HMBC), revealed the presence of [3-13C]serine and [13C- methyl]methionine. Label was also identified in a cysteinyl derivative and in several methoxylated compounds, but no evidence was obtained with either NMR or ion-trap mass spectrometry for the presence of any intermediate between nicotine and nornicotine. However, experiments with (R,S)-[14C- methyl]nicotine indicated that 70-75% of the metabolised label was released as carbon dioxide. These results are consistent with a pathway in which the oxidative hydrolysis of the nicotine methyl produces an unstable intermediate, N'-hydroxymethylnornicotine, that breaks down spontaneously to nornicotine and formaldehyde, with the formaldehyde being metabolised either directly to formate and carbon dioxide, or through the tetrahydrofolate-mediated pathways of one-carbon metabolism. However since the key intermediate, N-hydroxymethylnornicotine, could not be detected, the possibility of a direct methyl group transfer to tetrahydrofolate cannot be excluded. PMID- 11941470 TI - Tropisms in Phycomyces: sine law for gravitropism, exponential law for photogravitropic equilibrium. AB - Sporangiophores of Phycomyces blakesleeanus that are gravitropically stimulated by inclining them relative to the earth's gravitational vector obey the sine law for inclination angles between 0 degrees and 150 degrees. The quantitative relation between gravitropism and phototropism was analyzed for sporangiophores that were kept in balance between opposing gravitational and phototropic stimuli. The gravitropism of inclined sporangiophores was compensated with unilateral light impinging at right angles relative to the axis of the sporangiophore. The fluence rate of unilateral blue light (466 nm) that was required to counteract the negative gravitropism increased exponentially with the sine of the inclination angle of the sporangiophore. The establishment of photogravitropic equilibrium during continuous unilateral irradiation is thus determined by two different laws: the well-known sine law for gravitropism and a novel exponential law of phototropism described in this work. Furthermore, the specific form of the exponential relationship depends on the presence of statoliths (vacuolar protein crystals) and on wavelength. PMID- 11941469 TI - Daylength and spatial expression of a gibberellin 20-oxidase isolated from hybrid aspen (Populus tremula L. x P. tremuloides Michx.). AB - Physiologically active gibberellins (GAs) are key regulators of shoot growth in trees. To investigate this mechanism of GA-controlled growth in hybrid aspen, we cloned cDNAs encoding gibberellin 20-oxidase (GA 20-oxidase), a key, highly regulated enzyme in the biosynthesis of GAs. Clones were isolated from leaf and cambium cDNA libraries using probes generated by polymerase chain reaction, based on conserved domains of GA 20-oxidases. Upon expression in Escherichia coli, the GST-fusion protein was shown to oxidise GA12 as well as oxidising the 13 hydroxylated substrate GA53, successively to GA9 and GA20, respectively. The gene PttGA20ox1 was expressed in meristematic cells and growing tissues such as expanding internodes, leaves and roots. The expression was negatively regulated by both GA4 and overexpression of phytochrome A. RNA analysis also showed that the expression was down-regulated in late-expanding leaf tissue in response to short days (SDs). Actively growing tissues such as early elongating internodes, petioles and leaf blades had the highest levels of C19-GAs. Upon transfer to SDs an accumulation of GA19 was observed in early elongating internodes and leaf blades. The levels of C19-GAs were also to some extent changed upon transfer to SDs. The levels of GA20 were down-regulated in internodes, and those of GA1 were significantly reduced in early expanding leaf blades. In roots the metabolites GA19 and GA8 decreased upon shifts to SDs, while GA20 accumulated slightly. The down-regulation of GA 20-oxidase activity in response to SDs was further indicated by studies of [14C]GA12 metabolism in shoots, demonstrating that the substrate for GA 20-oxidase, [14C]GA53, accumulates in SDs. PMID- 11941471 TI - Expression of a vegetative-storage-protein gene from Arabidopsis is regulated by copper, senescence and ozone. AB - Emerging data suggest that the mechanisms regulating plant copper homeostasis could be implicated in stress and senescence signal transduction pathways. To gain insight into copper-modulated patterns of gene expression, copper-treated Arabidopsis thaliana (L.) Heynh. plants were analysed by mRNA differential display. The experimental conditions were selected using aggregation of ribulose 1,5-bisphosphate carboxylase-oxygenase (Rubisco) as a molecular sensor to monitor copper-induced oxidative stress. Two copper-induced messengers encoding a vegetative storage protein (VSP2) were isolated by this technique. Both clones differed in the length of their 3'-untranslated region according to the presence of two polyadenylation signals in this region. VSP2 expression was further studied under natural senescence and various conditions causing oxidative stress, such as ozone exposure, paraquat and H2O2 treatments. The expression of other messengers related to copper homeostasis and detoxification processes was followed in parallel to that of VSP2. Here, we describe specific gene-expression responses to copper treatment, and present arguments connecting copper homeostasis, senescence and antioxidative responses in plants. Our results are consistent with the role of VSPs as temporary nitrogen-storage proteins which accumulate if nutrients are abundant, either in developing organs or in cotyledons and mature leaves subjected to generalized protein mobilization, such as those conditions created under severe oxidative stress. PMID- 11941472 TI - The energy source for CO2 transport in the marine microalga Nannochloris atomus. AB - CO2 fluxes in the marine microalga Nannochloris atomus were studied by mass spectrometry using inhibitors and artificial acceptors of photosynthetic electron transport to investigate the energy source for CO2 uptake. This algal species is capable of taking up CO2 from the external medium by active transport but lacks active HCO(3)(-) transport and extracellular carbonic anhydrase. The capacity of cells to take up CO2 was a function of photosynthetic photon flux density. Dark respiration rates were also dependent upon the light intensity during the preceding illumination period, indicating the presence of light-enhanced dark respiration. Addition of 3-(3',4'-dichlorophenyl)-1,1-dimethylurea to illuminated cell suspensions that had been allowed to concentrate inorganic carbon internally during photosynthesis caused a rapid burst of CO2, demonstrating that active CO2 transport had been abolished. A similar response was obtained when cell suspensions were treated with 2,5-dibromo-6-isopropyl-3methyl-1,4-benzoqinone or hydroxylamine. When methyl viologen was used to drain electrons from ferredoxin, cells were still able to take up CO2 from the external medium, although C fixation decreased with time. These results demonstrate that active CO2 transport in N. atomus is supported by photosynthetic linear electron transport. PMID- 11941473 TI - Antisense-inhibition of ADP-glucose pyrophosphorylase in Vicia narbonensis seeds increases soluble sugars and leads to higher water and nitrogen uptake. AB - We previously reported on Vicia narbonensis seeds with largely decreased alpha- D glucose-1-phosphate adenyltransferase (AGP; EC 2.7.7.27) due to antisense inhibition [H. Weber et al. (2000) Plant J 24:33-43]. In an extended biochemical analysis we show here that in transgenic seeds both AGP activity and ADP-glucose levels were strongly decreased but starch was only moderately reduced and contained less amylose. The flux control coefficient of AGP to starch accumulation was as low as 0.08, i.e. AGP exerts low control on starch biosynthesis in Vicia seeds. Mature cotyledons of antisense seeds had increased contents of lipids, nitrogen and sulfur. The protein content was higher due, in particular, to increased sulfur-rich albumins. Globulin fractions of storage proteins had a lower ratio of legumin to vicilin. Isolated cotyledons partitioned less [14C]sucrose into starch and more into soluble sugars with no change in the protein fraction. Respiration of isolated cotyledons and activities of the major glycolytic and carbohydrate-metabolizing enzymes were not affected. Sucrose and the hexose-phosphate pool were increased but UDP-glucose, 3-phosphoglyceric acid, phospho enolpyruvate, pyruvate, ATP and ADP were unchanged or even lower, indicating that carbon partitioning changed from starch to sucrose without affecting the glycolytic and respiratory pathways. Soluble compounds were increased but osmolality remained unchanged, indicating compensatory water influx resulting in higher water contents. Developmental patterns of water and nitrogen accumulation suggest a coupled uptake of amino acids and water into cotyledons. We conclude that, due to higher water uptake, transgenic cotyledons take up more amino acids, which become available for protein biosynthesis leading to a higher protein content. Obviously, a substantial part of amino acid uptake into Vicia seeds occurs passively and is osmotically controlled and driven by water influx. PMID- 11941474 TI - Salinity-induced glutathione synthesis in Brassica napus. AB - The role of S-assimilation and the biosynthesis of cysteine and glutathione were studied during the response to salt stress of wild-type and salt-tolerant transgenic Brassica napus L. (canola) plants overexpressing a vacuolar Na+/H+ antiporter. A 3-fold increase in cysteine and glutathione content was observed in wild-type plants exposed to salt stress, but not in the transgenic plants. The induction of cysteine and glutathione synthesis during salt stress in the wild type plants suggests a possible protective mechanism against salt-induced oxidative damage. On the other hand, the salt-tolerant transgenic plants did not show significant changes in either cysteine or glutathione content, confirming the role of vacuolar Na+ accumulation and ion homeostasis in salt tolerance. PMID- 11941475 TI - In situ localisation of cytokinins in the shoot apical meristem of Sinapis alba at floral transition. AB - In plants of Sinapis alba L. induced to flower by one long day (LD), previous work showed that the phloem sap feeding the shoot apex is enriched in cytokinins of the isopentenyladenine (iP)-type between 9 and 25 h after start of the LD [P. Lejeune et al. (1994) Physiol Plant 90:522-528]. We have checked the hypothesis that the cytokinin content of the shoot apical meristem (SAM) should increase in response to floral induction by one LD using histoimmunolocalisation techniques and rabbit antiserum against isopentenyladenosine or zeatin riboside. The free bases iP and zeatin are present only in apical tissues containing dividing cells. At 30 h after the start of an inductive LD, a markedly increased iP immune reaction is observed in SAM tissues while the level of zeatin is not modified. Our results are in line with the data obtained by analysis of phloem sap. PMID- 11941477 TI - Mutations in PATCHED-1, the receptor for SONIC HEDGEHOG, are associated with holoprosencephaly. AB - Holoprosencephaly (HPE) is the most commonly occurring congenital structural forebrain anomaly in humans. HPE is associated with mental retardation and craniofacial malformations. The genetic causes of HPE have recently begun to be identified, and we have previously shown that HPE can be caused by haploinsufficiency for SONIC HEDGEHOG ( SHH). We hypothesize that mutations in genes encoding other components of the SHH signaling pathway could also be associated with HPE. PATCHED-1 (PTCH), the receptor for SHH, normally acts to repress SHH signaling. This repression is relieved when SHH binds to PTCH. We analyzed PTCH as a candidate gene for HPE. Four different mutations in PTCHwere detected in five unrelated affected individuals. We predict that by enhancing the repressive activity of PTCH on the SHH pathway, these mutations cause decreased SHH signaling, and HPE results. The mutations could affect the ability of PTCH to bind SHH or perturb the intracellular interactions of PTCH with other proteins involved in SHH signaling. These findings further demonstrate the genetic heterogeneity associated with HPE, as well as showing that mutations in different components of a single signaling pathway can result in the same clinical condition. PMID- 11941478 TI - Unstable transmission of the RS447 human megasatellite tandem repetitive sequence that contains the USP17 deubiquitinating enzyme gene. AB - The RS447 megasatellite DNA, which maps to human chromosome 4p16.1, is a highly polymorphic conserved tandem repetitive sequence containing a functional deubiquitinating enzyme gene, USP17. To characterize the hypervariability seen in RS447 fully, we have conducted a pedigree analysis of RS447 transmission by high resolution pulsed-field gel electrophoresis. We have identified 44 distinct alleles in 74 unrelated chromosomes containing 20-103 copies of the 4.7-kb RS447 unit. Five of 60 parent-to-offspring transmissions clearly show changes in copy number, indicating a high frequency (approximately 8.3%) of meiotic instability. Evidence for somatic mosaicism has also been observed. Searches of the database have revealed the presence of minor RS447 sequences mapping to chromosome 8p23, raising the possibility of a rearrangement or transposition of RS447 within the human genome. These results suggest that the unstable nature of RS447 megasatellite DNA gives rise to its hypervariability and may contribute to the structural dynamics of this repetitive DNA in the genome. PMID- 11941479 TI - Tandem duplication of the NF1 gene detected by high-resolution FISH in the 17q11.2 region. AB - The gene for neurofibromatosis type 1 (NF1), mapping to 17q11.2, has one of the highest observed mutation rates, partially because of its large size and gene conversion primed by NF1 pseudogenes. We have previously shown by means of high resolution fluorescence in situ hybridization (FISH) that a number of the loci flanking the NF1 gene are duplicated, in agreement with the reported presence of NF1 repetitive sequences (REPs). We report a direct tandem duplication of the NF1 gene identified in 17q11.2 by high-resolution FISH. FISH on stretched chromosomes with locus-specific probes revealed the duplication of the NF1 gene from the promoter to 3'UTR, but with at least the absence of exon 22. Fiber FISH with P1 artificial and bacterial artifical chromosomes, including the NF1 5'UTR and 3'UTR and flanking regions, visualized the direct tandem duplication with a similar, but not identical, genomic organization of the NF1 duplicon copies. Duplication was probably present in the human-chimpanzee-gorilla common ancestor, as demonstrated here by the finding of the duplicated NF1 gene at orthologous chromosome loci. The NF1 intrachromosomal duplication may contribute to the high whole-gene mutation rate by gene conversion, although the functional activity of the NF1 copy remains to be investigated. Detection of the NF1 duplicon by high resolution FISH may pave the way to filling the gaps in the human genomic sequence of the pericentromeric 17q11.2 region. PMID- 11941480 TI - Molecular characterization of a ring X chromosome in a male with short stature. AB - We report the molecular characterization of a ring X chromosome that was transmitted from a mother to a male who has short stature and minor dysmorphic features. This represents only the second reported ring X chromosome in a male. The ring is derived from breakage within the Xp pseudoautosomal region (PAR) and just proximal to the Xq PAR. The total amount of deleted material is 700-900 kb DNA and includes six known transcribed genes. Interestingly, SHOX, a gene implicated in short stature, is not deleted from the ring chromosome. Possible pathogenetic explanations for the patient's clinical features include insufficient dosage of deleted genes, a position effect on SHOX expression, and cell death during development because of ring chromosome nondisjunction. The findings are also relevant to observations made of "complete" ring chromosomes. PMID- 11941481 TI - Structure of the human argininosuccinate synthetase gene and an improved system for molecular diagnostics in patients with classical and mild citrullinemia. AB - Deficiency of argininosuccinate synthetase (ASS) causes citrullinemia, an autosomal recessive inherited defect of the urea cycle. Most patients described so far have presented with the classical form of the disease. There are also patients with a mild form of citrullinemia in whom the exact molecular basis and clinical relevance are uncertain. Mutations in the human ASS gene have not yet been described in mildly affected or asymptomatic patients with citrullinemia. The genomic sequence of the human ASS gene is not precisely known making mutation analysis difficult. Here, the entire genomic DNA sequence and mutations in the ASS gene of patients with the classical and mild form of the disease are described. The mutations c.1168G-->A (G390R) and IVS13+5 G-->A and the novel mutation c.323G-->T (R108L) have been found to be associated with classical citrullinemia, whereas the novel mutations c.535T-->G (W179R), and c.1085G-->T (G362V) have been detected on alleles of the mildly affected patients. Thus, mutations found in the human ASS gene of asymptomatic children with biochemical abnormalities and in some cases enzymatically proven citrullinemia have allowed us to classify these cases as ASS-deficient patients. The elucidation of the structure of the human ASS gene has made possible the use of intronic primers for molecular analysis of patients with mild disease and the classical form, and provides another option for prenatal diagnostics in affected families with the severe type. PMID- 11941482 TI - Vitiligo: complex segregation and linkage disequilibrium analyses with respect to microsatellite loci spanning the HLA. AB - Familial clustering and linkage disequilibrium studies suggest that genetic factors predispose to vitiligo, although a clear transmission pattern and cosegregation of vitiligo with specific mutations have not been demonstrated. We collected pedigree data on vitiligo from a set of 56 multigeneration families belonging to the Paisa community from Antioquia, Colombia, with the goal of applying the unified model of complex segregation and linkage disequilibrium analyses to test the hypotheses of the existence of a major gene predisposing to vitiligo and that allelic or haplotype polymorphisms of microsatellite loci at 6p21.3-21.4 spanning HLA (D6S276, D6S265, D6S273, and D6S291) are associated with this predisposition. Minimum sibship sample size to discriminate dominant and recessive inheritance models was largely accomplished. Between the 15 models of complex segregation used, the one that best fitted the data was that of a major dominant gene and the existence of strong environmental effects acting on the recessive genotype. The penetrance and risk estimations discriminated two sets of vitiligo patients: those with early onset of vitiligo cosegregating with a dominant mode of inheritance without environmental effects, and those with late onset of vitiligo cosegregating with the recessive genotype and being influenced by environmental effects. After establishing the normal distribution of allelic frequencies and performing multiple comparisons correction, the linkage disequilibrium analysis suggested that a major genetic factor could be located at 6p21.3-21.4, because we detected significant case-control differences for allele 122 at D6S265 ( Pc=0.0264) and significant linkage disequilibrium between loci D6S276 and D6S273 in the cases but not in the controls. We cannot explain these results as a consequence of evolutionary forces or as genetic stratification acting differentially on cases and controls, because there was neither deviation from the Hardy-Weinberg expectations nor genetic subdivision between cases and controls, as theta; (non-biased F(ST)) was not significantly different from 0. PMID- 11941483 TI - A locus for hereditary capillary malformations mapped on chromosome 5q. AB - Capillary malformations (port-wine stains) are the most common vascular malformations occurring in 0.3% of live births. Most capillary malformations occur sporadically and present as a solitary lesion. Capillary malformations can also occur as a component of well-described syndromes. Familial occurrence of multiple capillary malformations has been described in the literature, suggesting autosomal dominant inheritance with variable expression in this subgroup. A hereditary basis underlying the development of solitary capillary malformations has not been found, but may well be possible. We have mapped a locus for an autosomal dominant disorder in a three-generation family that manifested itself with multiple cutaneous capillary malformations to chromosome 5q13-22. This locus spans 48 cM between the markers D5S647 and D5S659 and harbours several candidate genes. By defining the gene(s) responsible for capillary malformations, we will gain more insight in the pathogenesis of this disorder. It is likely that genes implicated in these familial cases may be involved in the more sporadic cases. PMID- 11941484 TI - A novel locus for Usher syndrome type I, USH1G, maps to chromosome 17q24-25. AB - Usher syndrome (USH) is an autosomal recessive disorder associated with sensorineural hearing impairment and progressive visual loss attributable to retinitis pigmentosa. This syndrome is both clinically and genetically heterogeneous. Three clinical types have been described of which type I (USH1) is the most severe. Six USH1 loci have been identified. We report a Palestinian consanguineous family from Jordan with three affected children. In view of the combination of profound hearing loss, vestibular dysfunction, and retinitis pigmentosa in the patients, we classified the disease as USH1. Linkage analysis excluded the involvement of any of the known USH1 loci. A genome-wide screening allowed us to map this novel locus, USH1G, in a 23-cM interval on chromosome 17q24-25. The USH1G interval overlaps the intervals for two dominant forms of isolated hearing loss, namely DFNA20 and DFNA26. Since several examples have been reported of syndromic and isolated forms of deafness being allelic, USH1G, DFNA20, and DFNA26 might result from alterations of the same gene. Finally, a mouse mutant, jackson shaker ( js), with deafness and circling behavior has been mapped to the murine homologous region on chromosome 11. PMID- 11941485 TI - Contribution of arylsulfatase A mutations located on the same allele to enzyme activity reduction and metachromatic leukodystrophy severity. AB - The occurrence of different mutations on the same arylsulfatase A allele is not uncommon, due to the high frequency of several variants, among which the pseudodeficiency mutations are particularly important. We identified a late infantile metachromatic leukodystrophy patient carrying on one allele the new E253K mutation and the known T391S polymorphism, and on the other allele the common P426L mutation, usually associated with the adult or juvenile form of the disease, and the N350S and *96A>G pseudodeficiency mutations. To analyze the contribution of mutations located on the same allele to enzyme activity reduction, as well as the possible phenotype implications, we performed transient expression experiments using arylsulfatase A cDNAs carrying the identified mutations separately and in combination. Our results indicate that mutants containing multiple mutations cause a greater reduction of ARSA activity than do the corresponding single mutants, the total deficiency likely corresponding to the sum of the reductions attributed to each mutation. Consequently, each mutation may contribute to ARSA activity reduction, and, therefore, to the degree of disease severity. This is particularly important for the alleles containing a disease-causing mutation and the pseudodeficiency mutations: in these alleles pseudodeficiency could play a role in affecting the clinical phenotype. PMID- 11941487 TI - Homozygosity mapping of a Weill-Marchesani syndrome locus to chromosome 19p13.3 p13.2. AB - Weill-Marchesani syndrome (WMS) is a rare disease characterized by short stature, brachydactyly, joint stiffness, and characteristic eye abnormalities, including microspherophakia, ectopia lentis, and glaucoma. Both autosomal recessive and autosomal dominant modes of inheritance have been described in association with WMS. We have performed a genome-wide search in two large consanguineous families of Lebanese and Saudian origin consistent with an autosomal recessive mode of inheritance. Here, we report the linkage of the disease gene to chromosome 19p13.3-p13.2 (Zmax=5.99 at theta=0 at locus D19S906). A recombination event between loci D19S905 and D19S901 defines the distal boundary, and a second recombination event between loci D19S221 and D19S840 defines the proximal boundary of the genetic interval encompassing the WMS gene (12.4 cM). We hope that our ongoing studies will lead to the identification of the disease-causing gene. PMID- 11941486 TI - Genetic variation in alpha(1)-antichymotrypsin and its association with Alzheimer's disease. AB - Alzheimer's disease (AD) is a devastating neurodegenerative disorder characterized by extracellular neuritic plaques and intracellular neurofibrillary tangles in brain parenchyma. Alpha-1-antichymotrypsin (ACT) is a component of plaque cores, can bind to Abeta, and has been proposed as a possible candidate gene for AD susceptibility. The genetic association between the ACT codon -17*A allele of the signal peptide polymorphism and AD has been shown in some, but not in all studies. One hypothesis is that the ACT codon -17*A allele is in linkage disequilibrium with unknown functional mutation(s) in the ACT gene. This study was undertaken to identify new mutation(s) in the ACT gene by PCR-SSCP-sequencing and, in conjunction with known mutations, to assess their role in affecting the risk of AD. A total of seven new point mutations were observed: 5'UTR(A-->G), Asp128Asn(G-->A), Ser250Ser(C-->T), Leu301Pro(T-->C), Thr324Thr(A-->G), G-->A in intron 4, and 3'UTR C-->A. Of these, mutations at codon 250, codon 324, intron 4 and 3'UTR showed a frequency of 1% or more. Of the known mutations, Thr-17Ala(A- >G), Lys76Lys(A-->G) and Leu241Leu(G-->A) occur at a polymorphic level. The ACT codon -17*A allele was associated with increased risk of AD (OR for AA vs TT: 1.71; 95% CI: 1.16-2.53; P=0.007), especially in the presence of the APOE*4 allele (OR for AA vs TT: 2.35; 95% CI: 1.13-4.85; P=0.02). The codon 241*A allele and the codon 250*T allele were associated with protective effects against AD (OR: 0.36; 95% CI: 0.13-0.86; P=0.02) (OR:0.39; 95% CI: 0.18-0.85; P=0.02). irrespective of the APOE*4 status. The codon 324*G allele was associated with a marginal protective effect (OR:0.57; 95% CI: 0.26-1.26; P=0.17). While the codon 241*A allele was in linkage disequilibrium with the codon -17*A allele, the codon 250*T and codon 324*G alleles were non-randomly associated with the codon -17*T allele. In contrast, the codon 76*G (OR:1.34; 95% CI: 0.92-1.95; P=0.13), codon 227*G (OR:3.96; 95% CI: 0.83-18.8; P=0.08) and intron 4*G (OR:1.47; 95% CI: 0.88 2.29; P=0.15) alleles were associated with a modest risk of AD, and all were in linkage disequilibrium with the codon -17*A allele. EH-based haplotype analysis showed that certain haplotypes are associated with either higher or lower risk of AD. Our data indicate that the ACT gene harbors several potentially important variable sites, which are associated with either an increased or decreased risk of AD. The non-random combination of risk and protective alleles may explain, in part, why the association studies regarding the ACT codon -17*A have been inconsistent, especially if the frequency of other ACT mutations varies between populations. PMID- 11941488 TI - Haploinsufficiency of PAX9 is associated with autosomal dominant hypodontia. AB - We recently identified a frame-shift mutation in the PAX9 gene as the underlying cause for hypodontia involving permanent molar teeth segregating in an autosomal dominant pattern in a single large family (Stockton et al. 2000). Here we report a small nuclear family in which a father and his daughter are affected with severe hypodontia, involving agenesis of all primary and permanent molars, evidently caused by deletion of the entire PAX9 gene. Hemizygosity at the PAX9 locus in the two affected individuals was initially discovered when an informative single nucleotide polymorphism, identified while sequencing the gene for mutations, appeared to demonstrate non-Mendelian inheritance. Fluorescence in situ hybridization (FISH) analysis with a cosmid containing the PAX9 gene yielded a signal on only one chromosome 14 homologue and confirmed the presence of a deletion encompassing the PAX9 locus. Analysis of microsatellite loci in the vicinity of PAX9 delineated one breakpoint of the deletion. These data, in concert with FISH analysis with cosmids encompassing a 199 kb region, indicated that the deletion is between approximately 44 kb and 100 kb. PAX9 is one of two genes, and the only odontogenic gene within the deletion interval, thus supporting the model of haploinsufficiency for PAX9 as the underlying basis for hypodontia. PMID- 11941489 TI - The left-right determinant inversin has highly conserved ankyrin repeat and IQ domains and interacts with calmodulin. AB - All vertebrates have a left-right body axis with invariant asymmetries of the heart and the positions of the abdominal viscera. Major advances have recently been made in defining molecular components of the pathway specifying the vertebrate left-right axis, but our knowledge of the early determinants is extremely limited. In the invmouse the left-right axis is consistently reversed, unlike other vertebrate mutants where randomisation of situs is apparent. The gene disrupted in this mouse encodes a 1062-amino-acid protein, inversin. We previously reported 16 tandem ankyrin repeats, spanning amino acids 13-557, and two putative nuclear localisation sequences, but otherwise the sequence offered few clues to the possible function. In order to identify regions likely to be functionally important, we have identified and characterised orthologous sequences in several species, including chick, Xenopus and zebrafish. Sequence comparisons show strong conservation of the ankyrin repeat region and also a lysine-rich domain spanning amino acids 558-604. Further analysis identified a highly conserved IQ calmodulin-binding domain in the latter region and another such domain in an otherwise poorly conserved C-terminal region. A yeast two hybrid screen identified calmodulin in one third of the positive clones, and we confirmed this interaction by immunoprecipitation. PMID- 11941493 TI - Formation of a linear [3Fe-4S] cluster in a seven-iron ferredoxin triggered by polypeptide unfolding. AB - Cubic iron-sulfur ([Fe-S]) clusters are common inorganic cofactors in proteins. The presence of a linear [3Fe-4S] cluster in a protein was first observed in beef heart aconitase at high pH, where the protein structure was perturbed. Not long ago, the same linear cluster was discovered upon unfolding of a thermophilic di cluster seven-iron ferredoxin, suggesting a more general relevance for this type of linear clusters in Nature. Since structure-induced cluster rearrangements may be important regulatory, on-going processes in living systems, we decided to further characterize the formation of the linear iron-sulfur cluster observed upon ferredoxin unfolding. Here we present a kinetic investigation of parameters that affect the linear-cluster formation and disassembly in the Sulfolobus acidocaldarius seven-iron ferredoxin. We find the linear cluster to be an intermediate on the protein-mediated cluster-degradation pathway under a wide range of pH and denaturant conditions. The linear species forms in parallel with secondary-structure disappearance. In contrast, the disassembly rate constant for the linear cluster is independent of denaturant concentration but depends strongly on solution pH. At high pH, the disassembly rate is slower and the linear iron-sulfur species has a longer lifetime, than at low pH. PMID- 11941494 TI - Model investigations for vanadium-protein interactions: vanadium(III) compounds with dipeptides and their oxovanadium(IV) analogues. AB - The reaction of VCl(3) with 1,10-phenanthroline and a series of dipeptides (H(2)dip), having aliphatic as well as aromatic side chains, in methyl alcohol and in the presence of triethylamine affords vanadium(III) compounds of the general formula [V(III)(dip)(MeOH)(phen)]Cl. Aerial oxidation/hydrolysis of the vanadium(III) species gives their oxovanadium(IV) analogues of the general formula [V(IV)O(dip)(phen)]. X-ray crystallographic characterization of the [V(IV)O(dip)(phen)] compounds (where dip(2-)=Gly- L-Ala, Gly- L-Val and Gly- L Phe) revealed that the vanadium atom possesses a severely distorted octahedral coordination and is ligated to a tridentate dip(2-) ligand at the N(amine) atom, the deprotonated N(peptide) atom and one of the O(carboxylate) atoms, as well as an oxo group and two phenanthroline nitrogen atoms. Circular dichroism characterization of the V(III)/V(IV)O(2+)-dipeptide compounds revealed a strong signal for the V(IV)O(2+) species in the visible range of the spectrum, with a characteristic pattern which may be exploited to identify the N(am), N(pep) and O(car) ligation of a peptide or a protein to V(IV)O(2+) center, and a weak Cotton effect of opposite sign to their vanadium(III) analogues. The visible spectra of the V(III)-dipeptide compounds revealed two d-d bands with high intensity, thus indicating that the covalency of the metal-donor atoms is significant, i.e. the vanadium d orbitals are significantly mixed with the ligand orbitals, and this is confirmed by the low values of their Racah B parameters. The high-intensity band of the V(IV)O(2+)-dipeptide compounds at approximately 460 nm implies also a strong covalency of the metal with the equatorial donor atoms and this was supported by the EPR spectra of these compounds. Moreover, the V(III)/V(IV)O(2+) dipeptide complexes were characterized by EPR and IR spectroscopies as well as conductivity and magnetic susceptibility measurements. PMID- 11941495 TI - Kinetic studies on the oxidation of cytochrome b(5) Phe35 mutants with cytochrome c, plastocyanin and inorganic complexes. AB - To illustrate the functions of the aromatic residue Phe35 of cytochrome b(5) and to give further insight into the roles of the Phe35-containing hydrophobic patch and/or aromatic channel of cytochrome b(5), we studied electron transfer reactions of cytochrome b(5) and its Phe35Tyr and Phe35Leu variants with cytochrome c, with the wild-type and Tyr83Phe and Tyr83Leu variants of plastocyanin, and with the inorganic complexes [Fe(EDTA)](-), [Fe(CDTA)](-) and [Ru(NH(3))(6)](3+). The changes at Phe35 of cytochrome b(5) and Tyr83 of plastocyanin do not affect the second-order rate constants for the electron transfer reactions. These results show that the invariant aromatic residues and aromatic patch/channel are not essential for electron transfer in these systems. PMID- 11941496 TI - In vitro study of the insulin-mimetic behaviour of vanadium(IV, V) coordination compounds. AB - A representative set of vanadium(IV and V) compounds in varying coordination environments has been tested in the concentration range 1 to 10(-6) mM, using transformed mice fibroblasts (cell line SV 3T3), with respect to their short-term cell toxicity (up to 36 hours) and their ability to stimulate glucose uptake by cells. These insulin-mimetic tests have also been carried out with non transformed human fibroblasts (cell line F26). The compounds under investigation comprise established insulin-mimetic species such as vanadate ([H(2)VO(4)](-)), [VO(acetylacetonate)(2)], [VO(2)(dipicolinate)](-) and [VO(maltolate)(2)], and new systems and coordination compounds containing OO, ON, OS, NS and ONS donor atom sets. A vitality test assay, measuring the reduction equivalents released in the mitochondrial respiratory chain by intracellular glucose degradation, is introduced and the results are counter-checked with (3)H-labelled glucose. Most compounds are toxic at the 1 mM concentration level, and most compounds are essentially non-toxic and about as effective as or more potent than insulin at concentrations of 0.01 mM and below. V(V) compounds tend to be less toxic than V(IV)compounds, and complexes containing thio functional ligands are somewhat more toxic than others. Generally, ON ligation is superior in insulin-mimetic efficacy to OO or O/ NS coordination, irrespective of the vanadium oxidation state. There is, however, no striking correlation between the nature of the ligand systems and the insulin-mimetic potency in these cell culture tests, encompassing 41 vanadium compounds, the results on 22 of which are reported in detail here. The syntheses and characteristics of various new compounds are provided together with selected speciation results. The crystal and molecular structures of [[VO(naph-tris)](2)] [where naph-tris is the Schiff base formed between o-hydroxynaphthaldehyde and tris(hydroxymethyl)amine] are reported. Electronic supplementary material to this paper can be obtained by using the Springer Link server located at http://dx.doi.org/10.1007/s00775-001-0311-5. PMID- 11941497 TI - A comparison of DNA binding profiles of dinuclear platinum compounds with polyamine linkers and the trinuclear platinum phase II clinical agent BBR3464. AB - The DNA binding profiles of three bis Pt(II) polyamine-linked compounds, [[ trans PtCl(NH(3))(2)](2)[mu-spermine- N(1), N(12)]](4+), [[ trans-PtCl(NH(3))(2)](2)[mu spermidine- N(1), N(8)]](3+), and [[ trans-PtCl(NH(3))(2)](2)[mu-BOC spermidine]](2+), were compared with that of a novel trinuclear phase II clinical agent, [[ trans-PtCl(NH(3))(2)](2)[mu- trans Pt(NH(3))(2)(H(2)N(CH(2))(6)NH(2))(2)]](4+). All of the compounds bind preferentially in a bifunctional manner, according to unwinding of supercoiled DNA circles. The kinetics of binding of these compounds corresponds to their relative charge (2+ to 4+). The preference for the formation of interstrand crosslinks, however, does not follow a charge-based pattern. By studying the results of DNA polymerase extension products on a DNA template modified by the compounds, and by incorporating the complementary method of RNA transcription mapping, it was possible to determine the nucleotide bases that are preferred sites of binding. The stop sites due to platinum adducts were determined, and some preliminary observations concerning the range and type of crosslinks were established. It can be concluded that dinuclear Pt compounds are similar to their trinuclear counterpart, and that charge differences do not contribute solely to the variances between the compounds. PMID- 11941498 TI - Stabilities of lead(II) complexes formed in aqueous solution with methyl thiophosphate (MeOPS(2-)), uridine 5'- O-thiomonophosphate (UMPS(2-)) or adenosine 5'- O-thiomonophosphate (AMPS(2-)). AB - The acidity constants of twofold protonated methyl thiophosphate (MeOPS(2-)) and of monoprotonated uridine 5'- O-thiomonophosphate (UMPS(2-)) have been determined in aqueous solution (25 degrees C; I= 0.1 M, NaNO(3)) by potentiometric pH titration. The stability constants of their 1:1 complexes formed with Pb(2+), i.e. Pb(MeOPS) and Pb(UMPS), have also been measured. The results show that replacement of a phosphate oxygen by a sulfur atom increases the acidity by about 1.4 p K units. On the basis of recently established log versus plots ( = simple phosphate or phosphonate ligands where R is a non-coordinating residue), it is shown that the stability of the Pb(thiophosphate) complexes is by log Delta= 2.43+/-0.09 larger than expected for a Pb(2+)-phosphate interaction. The identity of the stability increase (log Delta) observed for Pb(MeOPS) and Pb(UMPS) shows that the nucleobase residue in the Pb(UMPS) complex has no influence on complex formation. To be able to carry out the mentioned comparisons, we have also determined the stability constant of the complex formed between Pb(2+) and methyl phosphate; the corresponding data for Pb(UMP) were already known from our earlier studies. The present results allow an evaluation of other Pb(2+) complexes formed with thiophosphate derivatives and they are applied now to the Pb(2+) complexes of adenosine 5'- O-thiomonophosphate (AMPS(2-)). The stability constants of the Pb(H;AMPS)(+) and Pb(AMPS) complexes were measured and it is shown that, within the error limits, the stability of the Pb(AMPS) complex is determined by the basicity of the thiophosphate group of AMPS(2-); in other words, no hint for macrochelate formation involving N7 was observed. More important, with the aid of micro-stability-constant considerations it is concluded that the structure of the dominating isomer of the Pb(H;AMPS)(+) species is the one where the proton is located at the N1 site of the adenine residue and Pb(2+) is coordinated to the deprotonated thiophosphate group. The insights gained from this study with regard to thiophosphate-altered single-stranded nucleic acids and their affinity towards Pb(2+) are discussed. PMID- 11941499 TI - Chemical shift-based constraints for solution structure determination of paramagnetic low-spin heme proteins with bis-His and His-CN axial ligands: the cases of oxidized cytochrome b(5) and Met80Ala cyano-cytochrome c. AB - The chemical shifts of the methyl protons of protoporphyrin IX, which are readily assigned, are related to the structural features of the axial histidine ligands in heme proteins with bis-His or His-CN axial coordination (Bertini I, Luchinat C, Parigi G, Walker FA (1999) JBIC 4:515-519). In the present paper, a module is developed which transforms the chemical shifts into a pseudo-potential energy that is a function of the dihedral angles defining the orientation of the axial ligand planes. Minimization of this pseudo-potential energy, together with the energetic contributions provided by the other constraints, yields structures consistent with the heme methyl chemical shifts. Oxidized cytochrome b(5) from rat and the cyanide derivative of the M80A mutant of yeast cytochrome c are used for test calculations. In the case of scarcity of NOEs for the axial ligands, owing to the presence of the paramagnetic center, the above structural constraints are shown to be quite precious. The newly refined structures are deposited in the PDB. PMID- 11941500 TI - Thermal stability of the [Fe(SCys)(4)] site in Clostridium pasteurianum rubredoxin: contributions of the local environment and Cys ligand protonation. AB - Thermal denaturation of the mesophilic rubredoxin from Clostridium pasteurianum occurs through a number of temperature-dependent steps, the last and irreversible one being release of iron from the [Fe(2+)(SCys)(4)] site. We show here that thermally induced [Fe(2+)(SCys)(4)] site destruction is largely determined by the local environment, and not directly connected to thermostability of the native polypeptide fold of rubredoxin. Hydrophobic residues on the protein surface, V8 and L41, that shield the [Fe(SCys)(4)] site from solvent and form N-H(.)S hydrogen bonds to the metal-coordinating sulfurs, were mutated to residues with both uncharged and charged side chains. On these mutated rubredoxins the temperature dependence was measured for: (1) global unfolding of the protein by NMR, (2) loss of Fe(2+)at various ionic strengths and pH values, (3) the rates of non-denaturing displacement of Fe(2+) by Cd(2+) or Zn(2+). For reversible temperature-dependent changes in the global protein folding that occur prior to loss of iron, no thermostability differences were found among the wild-type, V8A, V8D, L41R, and L41D rubredoxins. However, for irreversible loss of iron from the [Fe(2+)(SCys)(4)] site, relative to the wild-type protein, L41R was more thermostable, V8A was somewhat less thermostable, and the acidic mutants L41D, V8D and [V8D, L41D] showed dramatically lowered thermostability. Lower pH facilitated - both kinetically and thermodynamically - thermally induced iron release, likely through protonation of ligand cysteines' thiols. For all of the rubredoxins a direct correlation was found between the midpoint temperature for thermally induced Fe(2+) loss and the rate of non-denaturing Fe(2+) displacement by Cd(2+) or Zn(2+) at room temperature. A mechanism is proposed involving transient movement of residue-8 and -41 side chains, allowing, and, in the case of negatively charged side chains, also facilitating, attack of a ligand cysteine by the incoming positively charged species (H(+), Cd(2+), or Zn(2+)). Thus, localized charge density and solvent accessibility modulate the stability of Fe(2+) ligation in rubredoxin. However, the reduced [Fe(SCys)(4)] site does not control the thermostability of the native polypeptide fold of rubredoxin. PMID- 11941501 TI - A new urolith in four cats and a dog: composition and crystal structure. AB - A previously unreported urolith is presented. The concrement was found in the bladder of four Persian cats and one collie dog. It contained potassium, magnesium and phosphorus, all determined by chemical analysis. The infrared spectrum of the X-ray-dense urolith displayed absorption bands in the phosphate region. A single absorption maximum at 930 cm(-1) indicated the presence of inorganic pyrophosphate. A single crystal structure determination of the urolith has been performed. The results obtained are consistent with the formula [K(1.0)Mg(1.5)](4+)[P(2)O(7)](4-.)(H(2)O)(5), which corresponds to 11.5% K, 10.7% Mg and 18.2% P. The structure contains negatively charged slabs of composition MgP(2)O(7) small middle dot(H(2)O)(2)(2-)alternating with positively charged, hydrated Mg(2+) and K(+) ions. The observed P(2)O(7) is that of a distorted eclipsed syn conformation, which is the most common geometry in the solid state according to a statistical analysis of known structural data. A simple numerical descriptor has been derived, classifying observed X(2)O(7) conformations. It is suggested that some genetic enzymatic dysfunction of pyrophosphate-hydrolysing alkaline phosphatase might cause the formation of this unusual urolith. PMID- 11941502 TI - Magnetization studies of the active and fluoride-inhibited derivatives of the reduced catalase of Lactobacillus plantarum: toward a general picture of the anion-inhibited and active forms of the reduced dimanganese catalases. AB - The magnetic properties of the reduced catalase from Lactobacillus plantarum have been studied for the active enzyme and its fluoride complex through variable field/variable temperature magnetization measurements. The magnetic exchange interaction deduced from these experiments [fluoride complex: - J=1.3(1) cm(-1); active enzyme: - J=5.6(5) cm(-1); H=-2 J S(1) S(2)] are similar to those presently obtained in a re-analysis of the data for the corresponding forms of the Thermus thermophilus enzyme (previously published in 1997, Angew Chem Int Ed Engl 36:1626-1628): phosphate complex: - J=2.1(2) cm(-1); active enzyme - J=5.0(3) cm(-1). These results concur to a unified picture for the two enzymes, consistent with the presence of a hydroxide bridge in the reduced active catalases and its replacement by an aqua bridge in the anion-inhibited enzymes as the main mediators of the magnetic exchange. PMID- 11941503 TI - Stability and nickel binding properties of peptides designed as scaffolds for the stabilization of Ni(II)-Fe(4)S(4) bridged assemblies. AB - Helix-loop-helix peptides containing 63 residues (HC(4)H(2), HC(4)HC, HC(5)H), designated by their sequence and content of histidyl (H) and cysteinyl (C) residues, have been previously synthesized for the purpose of stabilizing certain bridged metal sites in proteins. These peptides bind one Fe(4)S(4) cluster by means of a ferredoxin tricysteinyl consensus sequence and an additional Cys residue, and one Ni(II) atom (HC(4)H(2), HC(5)H) in predesigned binding sites. In this investigation, the apopeptides and their Fe(4)S(4) derivatives are shown to be relatively stable to unfolding by guanidine hydrochloride, indicating stability of secondary structure. With this property demonstrated, Ni(II) binding equilibria have been evaluated in the terms of site-specific (Scatchard model) and stepwise (stoichiometric) binding constants. Two peptides were designed to have preformed CysHis(3) (HC(4)H(2)) and Cys(2)His(2) (HC(5)H) binding sites. The data indicate one strong binding site in each peptide with preferred binding constants k(1)=4.4x10(5) M(-1)(HC(4)H(2)) and 2.7x10(5) M(-1)(HC(5)H). Based on X ray absorption spectroscopic data, these binding steps are associated with the formation of the desired coordination units Ni(II)CysHis(3) and Ni(II)Cys(2)His(2). For peptide HC(4)HC, k(1)=2.5x10(5) M(-1), but the binding site could not be fully identified. Collective evidence from this and prior investigations supports the presence of the bridged assemblies Ni(II)-(mu(2)-S x Cys)-[Fe(4)S(4)], stabilized by a scaffolding effect in peptides HC(4)H(2) and HC(5)H. The assembly Ni(II)-X-[Fe(4)S(4)] is the minimal structure of the A Cluster of carbon monoxide dehydrogenase adduced from spectroscopic evidence; bridge X is currently unidentified. These results suggest that de novo designed peptides may serve as scaffolds for the construction of native bridged sites in proteins. Electronic supplementary material to this paper can be obtained by using the Springer Link server located at http://dx.doi.org/10.1007/s00775-001 0320-4. PMID- 11941504 TI - Solution structure of the unbound, oxidized Photosystem I subunit PsaC, containing [4Fe-4S] clusters F(A) and F(B): a conformational change occurs upon binding to photosystem I. AB - This work presents the three-dimensional NMR solution structure of recombinant, oxidized, unbound PsaC from Synechococcus sp. PCC 7002. Constraints are derived from homo- and heteronuclear one-, two- and three-dimensional (1)H and (15)N NMR data. Significant differences are outlined between the unbound PsaC structure presented here and the available X-ray structure of bound PsaC as an integral part of the whole cyanobacterial PS I complex. These differences mainly concern the arrangement of the N- and C-termini with respect to the [4Fe-4S] core domain. In the NMR solution structure of PsaC the C-terminal region assumes a disordered helical conformation, and is clearly different from the extended coil conformation, which is one of the structural elements required to anchor PsaC to the PS I core heterodimer. In solution the N-terminus of PsaC is in contact with the pre-C-terminal region but slides in between the latter and the iron-sulfur core region of the protein. Together, these features result in a concerted movement of the N-terminus and pre-C-terminal region away from the F(A) binding site, accompanied by a bending of the N-terminus. In comparison, the same terminal regions are positioned much closer to F(A) and take up an anti-parallel beta-sheet arrangement in PsaC bound to PS I. The conformational changes between bound and unbound PsaC correlate with the differences reported earlier for the EPR spectra of reduced F(A) and F(B) in bound versus unbound PsaC. The observed different structural features in solution are highly relevant for unraveling the stepwise assembly process of the stromal PsaC, PsaD and PsaE subunits to the PS I core heterodimer. Electronic supplementary material to this paper can be obtained by using the Springer Link server located at http://dx.doi.org/10.1007/s00775-001 0321-3. PMID- 11941505 TI - New potent agents binding to a poly(dT) sequence in double-stranded DNA: bis(Zn(2+)-cyclen) and tris(Zn(2+)-cyclen) complexes. AB - In an effort to search for mechanistically new and more potent agents than conventional drugs that target AT-rich sequences in double-stranded DNA, we have tested multi(Zn(2+)-cyclen) complexes. Indeed, they selectively bound to poly(dT) sequences to melt the A-T hydrogen bonds; only 2.5 microM or 4 microM of the p tris(Zn(2+)-cyclen) complex were required to completely melt a 50 microM nucleobase of double-stranded poly(dA) x poly(dT) or poly(dA-dT)(2) at 25 degrees C. The region with seven consecutive T's in native DNA (150 bp) was protected from micrococcal nuclease hydrolysis, as revealed by footprinting assays, with IC(50) values of 2 microM for p-bis(Zn(2+)-cyclen) and 0.5 microM for p tris(Zn(2+)-cyclen). The high affinity to AT-rich sequences of these Zn(2+) cyclen complexes matches or surpasses those of the conventional AT-binding drugs distamycin A (IC(50)=2 microM) and DAPI (5 microM). Moreover, the p-tris(Zn(2+) cyclen) complex selectively binds to the TATA box sequence of the SV40 early promoter to inhibit the binding of the TATA binding protein as effectively as distamycin A, with an IC(50) value of 0.4 microM. In vitro transcription of poly(dA) x poly(dT) using Escherichia coli RNA polymerase was effectively inhibited by p-tris(Zn(2+)-cyclen). The [(3)H]-ATP incorporation into RNA was more strongly blocked (IC(50)=0.8 microM) than the [(3)H]-UTP incorporation (IC(50)=40 microM), a fact indicating that the p-tris(Zn(2+)-cyclen) complex interacts only with the poly(dT) strand in the double-stranded DNA template. PMID- 11941506 TI - An archaeal b-type cytochrome containing a nonfunctional carbonic anhydrase-like domain. AB - A new type of cytochrome b was isolated from the cytoplasmatic fraction of the archaeon Acidianus ambivalens, which is the first soluble cytochrome found in this member of the thermoacidophilic order of the Sulfolobales. The protein is a monomeric and monohemic cytochrome b with a molecular mass of 22 kDa. Visible spectroscopy of the as-purified protein shows a Soret peak at 405 nm and a broad band at 625 nm, indicating the presence of a high-spin ferric heme. Upon reduction, the Soret band shifts to 422 nm and a broad band at 560 nm develops, again characteristic of high-spin ferrous heme. The reduced form can bind carbon monoxide, with visible absorption bands arising at 411 and 566 nm. EPR spectroscopy of the oxidized protein shows a spectrum typical of a high-spin heme, with major g values at 6.56 and 5.85. The reduction potential of the heme cofactor was determined to be -16+/-10 mV, at pH 6.5. Analysis of the protein amino acid sequence shows that it consists of a novel arrangement of domains. The first domain, at the N-terminus, has a remarkable similarity towards beta class carbonic anhydrases, whereas the second region comprises a putative cytochrome domain. The latter presumably consists of a novel fold, as it bears no sequence similarities towards other known cytochromes, or towards known domains. Strikingly, the first module contains the C-X (n)-H-X(2)-C motif that accounts for the binding of the catalytic zinc in carbonic anhydrases, but lacks several other critical residues required for substrate binding and proper active site geometry. In agreement with this finding, the isolated cytochrome contains one bound zinc atom, but has no carbonic anhydrase activity. Inspection of the sequences available from the genomic sequencing project of the close relative archaeon Sulfolobus solfataricus shows the presence of an identical protein, suggesting its dissemination among the Sulfolobales. The role of zinc as a key element for the intrinsic thermal stability of these proteins is discussed. PMID- 11941507 TI - Three high-resolution crystal structures of cadmium-substituted carboxypeptidase A provide insight into the enzymatic function. AB - Three high-resolution crystal structures of Cd(II)-substituted carboxypeptidase A (CPA) have been determined by X-ray diffraction from crystals prepared in three different buffer systems to assess the effect of pH and ionic strength on the Cd(II) coordination geometry. All crystallize in the space group P2(1) with identical cell dimensions. Cd-CPA(7.5): Cd(II)-substituted CPA prepared at pH 7.5 with [Cl(-)]=2 mM determined to 1.70 A resolution ( R=17.4% and R(free)=19.8%); Cd-CPA(5.5): Cd(II)-substituted CPA prepared at pH 5.5 with [Cl(-)]=2 mM to 2.00 A resolution ( R=16.1% and R(free)=18.6%); Cd-CPA(7.5)-Cl: Cd(II)-substituted CPA prepared at pH 7.5 with [Cl(-)]=250 mM to 1.76 A resolution ( R=16.7% and R(free)=17.8%). No noticeable structural changes were observed between the three structures. Two water molecules coordinate to Cd(II), in contrast to the single water molecule coordinating to Zn(II) in the Zn-CPA structure. No binding sites for anions could be identified, even in the structure with a high concentration of chloride ions. It is suggested that the anion inhibition is due to weak outer sphere association of Cl(-) ions at several binding sites, shielding the strong positive charge distribution at the surface of the protein near the active site. Based on structural data and a sequence alignment of 18 non-redundant carboxypeptidases, a more elaborate version of the earlier reaction model is proposed that also addresses the transport of water to and from the active site. Conserved residues whose function was not addressed previously delineate the proposed pathways used in the transport of water during catalysis. PMID- 11941508 TI - The nickel enzyme methyl-coenzyme M reductase from methanogenic archaea: In vitro induction of the nickel-based MCR-ox EPR signals from MCR-red2. AB - Methyl-coenzyme M reductase (MCR) is a nickel enzyme catalyzing the formation of methane from methyl-coenzyme M and coenzyme B in all methanogenic archaea. The active purified enzyme exhibits the axial EPR signal MCR-red1 and in the presence of coenzyme M and coenzyme B the rhombic signal MCR-red2, both derived from Ni(I). Two other EPR-detectable states of the enzyme have been observed in vivo and in vitro designated MCR-ox1 and MCR-ox2 which have quite different nickel EPR signals and which are inactive. Until now the MCR-ox1 and MCR-ox2 states could only be induced in vivo. We report here that in vitro the MCR-red2 state is converted into the MCR-ox1 state by the addition of polysulfide and into a light sensitive MCR-ox2 state by the addition of sulfite. In the presence of O(2) the MCR-red2 state was converted into a novel third state designated MCR-ox3 and exhibiting two EPR signals similar but not identical to MCR-ox1 and MCR-ox2. The formation of the MCR-ox states was dependent on the presence of coenzyme B. Investigations with the coenzyme B analogues S-methyl-coenzyme B and desulfa methyl-coenzyme B indicate that for the induction of the MCR-ox states the thiol group of coenzyme B is probably not of importance. The results were obtained with purified active methyl-coenzyme M reductase isoenzyme I from Methanothermobacter marburgensis. They are discussed with respect to the nickel oxidation states in MCR-ox1, MCR-ox2 and MCR-ox3 and to a possible presence of a second redox active group in the active site. Electronic supplementary material to this paper can be obtained by using the Springer LINK server located at http://dx.doi.org/10.1007/s00775-001-0325-z. PMID- 11941509 TI - Hybrid cluster proteins (HCPs) from Desulfovibrio desulfuricans ATCC 27774 and Desulfovibrio vulgaris (Hildenborough): X-ray structures at 1.25 A resolution using synchrotron radiation. AB - The structures of the hybrid cluster proteins (HCPs) from the sulfate-reducing bacteria Desulfovibrio desulfuricans (ATCC 27774) and Desulfovibrio vulgaris (Hildenborough) have been elucidated at a resolution of 1.25 A using X-ray synchrotron radiation techniques. In the case of the D. desulfuricans protein, protein isolation, purification, crystallization and X-ray data collection were carried out under strict anaerobic conditions, whereas for the D. vulgaris protein the conditions were aerobic. However, both structures are essentially the same, comprising three domains and two iron-sulfur centres. One of these centres situated near the exterior of the molecules in domain 1 is a cubane [4Fe-4S] cluster, whereas the other, located at the interface of the three domains, contains the unusual four-iron cluster initially found in the D. vulgaris protein. Details of the structures and the associated EPR spectroscopy of the D. desulfuricans protein are reported herein. These structures show that the nature of the hybrid cluster, containing both oxygen and sulfur bridges, is independent of the presence of oxygen in the isolation and crystallization procedure and also does not vary significantly with changes in the oxidation state. The structures and amino acid sequences of the HCP are compared with the recently elucidated structure of the catalytic subunit of a carbon monoxide dehydrogenase from Carboxydothermus hydrogenoformans and related dehydrogenases. Electronic supplementary material to this paper can be obtained by using the Springer Link server located at http://dx.doi.org/10.1007/s00775-001-0326-y. PMID- 11941510 TI - Iron-sulfur cluster biosynthesis: characterization of Schizosaccharomyces pombe Isa1. AB - Eukaryotic Isa1 is one of several mitochondrial proteins that have been implicated in Fe-S cluster assembly paths in vivo. We report the first biochemical characterization of an eukaryotic member of this family and discuss this in the context of results from in vivo studies and studies of bacterial homologues. Schizosaccharomyces pombe Isa1 is a multimeric protein carrying [2Fe 2S](2+) clusters that have been characterized by Mossbauer and optical spectroscopic studies. Complex formation with a redox-active ferredoxin has been identified through crosslinking experiments and the coordination chemistry and stability of the native clusters has been investigated through site-directed mutagenesis and spectroscopic analysis. Electronic supplementary material to this paper, containing Mossbauer and UV-visible spectra for mutant Isa1 proteins, can be obtained by using the Springer Link server located at http://dx.doi.org/10.1007/s00775-001-0330-2. PMID- 11941511 TI - Interaction of zinc ions with d(CGCAATTGCG) in a 2.9 A resolution X-ray structure. AB - We have synthesized and crystallized in the presence of Zn(2+) ions the peptidyl oligonucleotide adduct CH(3)CO-(Arg)(4)-NH-(CH(2))(6)-NH-p-d(CGCAATTGCG). This is the first structure obtained from a deoxyoligonucleotide crystallized in the presence of zinc ions. Zn ions are clearly visible in the 2.9 A resolution map. On the other hand, the peptide tail is not visible in the crystal structure as determined by X-ray diffraction. The terminal bases C1 and G10 are found in extra helical positions. Their phosphates are ligands of a Zn(2+) ion, located in a special position of the unit cell. This ion plays an important role in the packing arrangement, since it binds four different DNA molecules. Two other Zn(2+) ions are also important for DNA packing. They interact specifically with the N7 atoms of the terminal G2 and G10 bases, but not with the internal G8. This result supports the hypothesis that transition metals do not interact with the bases of duplex DNA in the B form. PMID- 11941512 TI - Solution structure of cyanoferricytochrome c: ligand-controlled conformational flexibility and electronic structure of the heme moiety. AB - The solution structure of cyanoferricytochrome c has been determined using NMR spectroscopy. As a result of including additional constraints derived from pseudocontact shifts, a high-resolution NMR structure was obtained with high accuracy. In order to study the conformational transition between the native protein and its ligand adducts, the present structure was compared with the solution structures of the wild-type cytochrome c and the imidazole-cytochrome c complex. Like the solution structure of imidazole-cytochrome c, the heme crevice is widened by the swinging out of residues 77-85 and a noticeable shift of the 50s helix. However, unlike imidazole, cyanide exerts less significant perturbation on the conformation of the heme cavity, which is revealed by a more compact residue package in the distal pocket. Furthermore, comparison of the solution structure of CN-iso-1Met80Ala cytochrome c with the structure of cyanoferricytochrome c indicated that the binding of cyanide has a different impact on the distal cavity conformation in the two proteins. In addition, the magnetic properties of the present system are discussed and a comprehensive study of the electronic structure of ligand-cytochrome c complexes and the native protein is also described. Electronic supplementary material to this paper can be obtained by using the Springer Link server located at http://dx.doi.org/10.1007/s00775-001-0334-y. PMID- 11941513 TI - The level of 8-oxo-7,8-dihydro-2'-deoxyguanosine is positively correlated with the size of the labile iron pool in human lymphocytes. AB - It appears that the labile iron pool (LIP, low molecular weight iron) presence in cells can result in the production of reactive oxygen species (ROS). ROS may be responsible for the formation of 8-oxo-7,8-dihydro-2'-deoxyguanosine (8-oxodGuo) in cellular DNA. In the present study we report on the relationship between LIP and the endogenous level of 8-oxodGuo in human lymphocytes. Good correlation has been determined between LIP and the oxidatively modified nucleoside. This in turn points out the possibility that under physiological condition there is the availability of LIP for catalyzing Fenton-type reactions in close proximity to cellular DNA. Electronic supplementary material to this paper can be obtained by using the Springer Link server located at http://dx.doi.org/10.1007/s00775-001 0335-x. PMID- 11941514 TI - Characterization of a metalloregulatory bismuth(III) site in Staphylococcus aureus pI258 CadC repressor. AB - Staphylococcus aureus pI258 CadC is a metal sensor protein that regulates the expression of the cad operon which encodes metal ion resistance proteins involved in the efficient efflux of Cd(II), Pb(II), Zn(II) and, according to one report, Bi(III) ions. In this paper, direct evidence is presented that Bi(III) binds to CadC and negatively regulates cad operator/promoter (O/P) binding. Optical absorption spectroscopy reveals that dimeric CadC binds approximately 0.8 mol equivalents of Bi(III) per CadC monomer to form a coordination complex characterized by three S(-)-->Bi(III) ligand-to-metal charge transfer transitions, with the longest wavelength absorption band centered at 415 nm (epsilon(415)=4000 M(Bi)(-1) cm(-1)). UV-Vis absorption spectra of wild-type and mutant Cys-->Gly (Ser) substitution CadC mutants compared to [Bi(DTT)(2)], [Bi(GSH)(3)] and [Bi(NAC)](3) model complexes reveal that Cys7, Cys11, Cys60 and Cys58 directly coordinate Bi(III) in a tetrathiolate coordination complex. The apparent affinity derived from a Bi(III)-displacement optical titration with Cd(II) is estimated to be K(Bi)< or =10(12) M(-1). Apo-CadC binds with high affinity [ K(a)=1.1(+/-0.3)x10(9) M(-1); 0.40 M NaCl, pH 7.0, 25 degrees C] to a 5'-fluorescein-labeled cad O/P oligonucleotide,while the binding of one molar equivalent of Bi(III) per CadC monomer (Bi(1)-CadC) reduces the affinity by approximately 170-fold. Strikingly, Bi(III)-responsive negative regulation of cad O/P binding is abrogated for Bi(1)-C60G CadC and severely disrupted in Bi(1)-C7G CadC, whose relative affinity is reduced only 10-fold. The mechanism of Bi(III) responsive metalloregulation is discussed, based on the findings presented here. Electronic supplementary material to this paper can be obtained by using the Springer Link server located at http://dx.doi.org/10.1007/s00775-001-0336-9. PMID- 11941518 TI - [Attachment and connections]. PMID- 11941519 TI - [The differential relevance of attachment classification for psychological disorders]. AB - In recent years, there is a growing acceptance of attachment theory in different fields of psychosocial medicine. This article reviews findings of clinical attachment research - mainly based upon the Adult Attachment Interview - related to three psychological disorders, i.e. Anxiety Disorders, Depression and Borderline Personality Disorder. Whilst there are some indications for a differentiation of de- and hyperactivating attachment strategies connected to depressive disorders, the findings related to anxiety and Borderline Personality Disorders clearly indicate a predominance of preoccupied attachment patterns combined unresolved traumatic experiences. The need for a further differentiation of attachment related features within clinical samples should be considered in the future. Specifically, a differentiation of "organized" vs. "disorganized" attachment representations seems to be promising. PMID- 11941520 TI - [Attachment characteristics and psychotherapy outcome following inpatient psychodynamic group treatment]. AB - Within a reanalysis of the "Kiel Group Psychotherapy Study", the videorecorded intake interviews of 34 patients were analyzed using the adult attachment prototype rating. Three subgroups, differentiated according to their attachment patterns (avoidant, ambivalent, mixed insecure) differed marginally with respect to their treatment success, with ambivalent patients showing the most positive outcome (escpecially with regard to symptom- and self-system-related outcome criteria). In addition, there were indications for a differential assessment of group related therapeutic factors by the subgroups. Patients with an avoidant attachment pattern evaluated group-related factors such as cohesion and altruism as least helpful. The study is discussed in relation to existing findings on the significance of attachment related variables for the indication and prognosis of psychotherapy. PMID- 11941521 TI - [The "Well-being paradox" in quality-of-life research]. AB - In quality-of-life research an empirical phenomenon has been observed independently in medicine and in the social sciences which is so contrary to what intuition tells us that it has been called paradoxical. In medicine it is called the "satisfaction paradox" and in the social sciences the "well-being paradox". What is meant in both cases is that objectively negative factors in one's life have relatively little effect on subjective quality of life. In the present paper examples are given of relevant research findings from both fields. An attempt is then made to explain the phenomenon, with the topic being examined not only from a methodological but also from a personality, cognition and social psychology perspective. Finally, the implications for research and clinical work are discussed. PMID- 11941522 TI - [Interpersonal psychotherapy (IPT) in depressions]. AB - Evidence-based medicine gains more importance within psychotherapy as well, although some problems occur in trying to follow the principles of evidence-based medicine in the field of psychotherapy very strictly. The Interpersonal Psychotherapy (IPT) for depression, which has been developed more than 30 years ago may be taken as an example for evidence-based-psychotherapy: the concept was built on empirical observations and manualised. Further on it was evaluated in many clinical trails that showed the good efficacy and later effectiveness. IPT reaches a high level of evidence and is to be seen as well-established psychotherapy. Psychotherapy research tries to point out the effectiveness for different subtypes of depressive disorders. Moreover it is investigated which connection exist between therapy outcome and neurobiological changes in the central nervous system. PMID- 11941523 TI - [Psycho-physiological aspects of gastroesophageal reflux disease]. AB - It is well known that there is an interaction between physiological and psychological aspects of gastrointestinal diseases, also in esophageal symptoms. Based on this bio-psycho-social interaction, several multidisciplinary concepts of interventions in gastrointestinal disorders have been evaluated. The role of psychological factors in gastroesophageal reflux disease (GERD) is really unknown. The present article reviews the basic pathophysiological factors of GERD including psycho-physiological aspects and presents potential concepts of multidisciplinary GERD treatment. PMID- 11941524 TI - [Autonomic neuropathy in somatization disorders]. AB - AIM: We conducted a study to investigate whether patients with somatization disorder show abnormal values in autonomic testing, especially in the central baroreceptor sensitivity. PATIENTS AND METHODS: Seventy-one patients were included. All had a diagnosis of somatization disorder (ICD-10, F45.0). Psychometric testing was performed by means of validated questionnaires (STAI, STAXI, FPI, GBB, ADS, SOMS, SCL-90-R). Autonomic regulation was analyzed by international standards using frequency spectral calculation by fast Fourier transformation. Thereby 3 different groups were detected: 12 patients with a baroreceptor sensitivity (BRS) of less than 3.0 ms/mm Hg, 20 patients with normal BRS (> 9.0 ms/mm Hg), and an in-between group (n = 39) with intermediate BRS. Controlling for age, a covariance analysis was calculated. RESULTS: The two extreme groups showed no difference in psychometric testing. However, significant differences were discernible in spectral values of mid-frequency-band (p < 0.05) in a covariance analysis with age as covariate. Equally the 24 h blood pressure determination showed significantly higher values for the group with BRS < 3.0 ms/mm Hg (p < 0.05 to 0.001). CONCLUSIONS: In a high percentage (17 %) of patients diagnosed to have somatization disorder autonomic dysregulation becomes apparent and is accompanied by increased blood pressure. Therefore it doesn't seem accurate to overlook concomitant organic lesions in somatization disorders despite patients lacking overtly clinical signs but suffering from various unspecific symptoms. PMID- 11941525 TI - [Importance of the therapeutic alliance experience for the differential psychotherapeutic indication]. AB - The influence of the therapeutic alliance experience on the recommendation for psychotherapy and prognosis was examined in a retrospective study. Three hundred seventy-four of 3270 patients who presented at the psychosomatic psychotherapeutic outpatient service at the Benjamin Franklin Hospital of the Free University of Berlin between July 1994 and December 1999 were selected based on the fact that they had been examined only by a psychoanalyst and recommended for individual psychoanalytical outpatient therapy, nondirective therapy (according to Rogers) or behavioral therapy. Results of the unifactorial variance analysis: The better the alliance is assessed, the sooner procedures for psychoanalytical therapy are recommended (F = 35.85; p < 0.000) and the better the assessment of the prognosis (r = 0.68; p < 0.000). The results confirm the importance of the therapeutic working relationship and correct the assumption that the indication and prognosis are exclusively diagnosis or patient-related, clearly showing that they are the result of an interactive process. PMID- 11941526 TI - [The relationship patterns questionnaire (RPQ): validation by using a representative sample]. AB - In this study the present form of the Relationship Patterns Questionnaire (RePat) will be presented as a successor of the Interpersonal Relationship Patterns Questionnaire (IRQ). Similar to the IRQ the RePat assesses the behavior of relationship concerning both one's own and the anticipated behavior of an important person on the basis of the SASB model and the CCRT method. In contrast to the IRQ, the RePat is much shorter and thus easier to use. The study describes the construction and the psychometric examination of the questionnaire by using a representative study (n = 2007, East = 1015, West = 992). The samples of East and West Germany show structural differences concerning its relationship patterns, which in our opinion are plausible for political and historical reasons. The clinical application and their implications for therapeutic interventions will be demonstrated by a single case using both the patient's and her partner's assessment of their relationship. PMID- 11941527 TI - [The German short version of the interpersonal guilt questionnaire--Validation in a population-based sample and clinical application]. AB - The present study describes the development of the German short version "Fragebogen zu interpersonellen Schuldgefuhlen" (FIS) of the "Interpersonal Guilt Questionnaire" (IGQ) developed by O'Connor et al. The questionnaire was used in a representative sample of 652 eastern and 1283 western Germans. Woman show higher scores on the subscales "Seperation Guilt" and "Omnipotent Responsibility Guilt" than man. Eastern Germans mark higher scores on the subscale "Omnipotent Responsibility Guilt" than western Germans. Correlations between interpersonal guilt and social anxiety and differences between a non-clinical and a clinical sample show that maladaptive aspects of guilt feelings can be assessed by the "Interpersonal Guilt Questionnaire". PMID- 11941530 TI - The history and principles of cryopreservation. AB - The ability of glycerol to protect cells from freezing injury was discovered accidentally. The subsequent development of cryopreservation techniques has had a huge impact in many fields, most notably in reproductive medicine. Freezing injury has been shown to have two components, direct damage from the ice crystals and secondary damage caused by the increase in concentration of solutes as progressively more ice is formed. Intracellular freezing is generally lethal but can be avoided by sufficiently slow cooling, and under usual conditions solute damage dominates. However, extracellular ice plays a major role in tissues. Cryoprotectants act primarily by reducing the amount of ice that is formed at any given subzero temperature. If sufficient cryoprotectant could be introduced, freezing would be avoided altogether and a glassy or vitreous state could be produced, but osmotic and toxic damage caused by the high concentrations of cryoprotectant that are required then become critical problems. The transport of cryoprotectants into and out of cells and tissues is sufficiently well understood to make optimization by calculation a practical possibility but direct experiment remains crucial to the development of other aspects of the cryopreservation process. PMID- 11941531 TI - Cryopreservation of spermatozoa in assisted reproduction. AB - Spermatozoa were the first cells to be cryopreserved over 50 years ago, following the serendipitous discovery of the cryoprotective compound glycerol. This pioneering work was followed by the introduction of a series of other cryoprotectant chemicals referred to collectively as cryoprotective agents. Glycerol has been widely used in the cryopreservation of bull and human spermatozoa, although results are still highly variable across species as well as among individuals within a species. Recently, significant information has been gained with regard to the fundamental cryobiology of several mammalian species' spermatozoa that can be used to reduce this variability and develop improved methods for cryopreservation. In this concise review, we will discuss the fundamental cryobiology of cells in general and of mammalian spermatozoa in particular. PMID- 11941532 TI - The use of cryopreserved mature and immature testicular spermatozoa for intracytoplasmic sperm injection: risks and limitations. AB - Treatment of severe male subfertility has become available since the intracytoplasmic injection of a single sperm into an oocyte was successfully applied for the first time in 1992. Moreover, with the use of fresh and cryopreserved epididymal and testicular spermatozoa for this procedure, fertilization and pregnancies could be accomplished. This review addresses the development and performance of these techniques and discusses achievements and problems as well as future aspects of the feasibility of early spermatid injection. Furthermore, limitations of these procedures and concerns with regard to genetic and epigenetic risks of using immature gametes are stressed. PMID- 11941533 TI - Cryopreservation, thawing, and transfer of human embryos. AB - Cryopreservation of human embryos is a routine procedure in most assisted reproductive technology laboratories. Embryos can be successfully frozen at the pronuclear, multicellular (four to eight blastomeres), and blastocyst stages. Each laboratory has to determine which freezing protocol is best suited for its patients. Freezing protocols have been developed that utilize cryoprotective agents such as glycerol, propanediol, dimethylsulfoxide, and sucrose. The purpose of these cryoprotectants is to help reduce the formation of intracellular ice when using a slow cooling protocol to freeze the embryos. The two most common ways to freeze embryos utilize either low concentrations of cryoprotectants and a slow stepwise freeze or high concentrations of cryoprotectants and a rapid freeze. Frozen-thawed embryos can be transferred to naturally cycling women or to women who have been primed with hormone replacement treatment with equal success. PMID- 11941534 TI - Cryopreservation of immature and mature human oocytes. AB - Cryopreservation of oocytes facilitates the long-term storage of oocytes for patients in danger of losing ovarian function. It also alleviates many of the ethical concerns associated with embryo cryopreservation. Problems associated with metaphase II oocyte cryopreservation include zona pellucida hardening and spindle damage. The cryopreservation of germinal vesicle-stage oocytes has been undertaken as a means of circumventing the problem of spindle damage in mature oocytes. One of the main disadvantages of immature oocyte cryopreservation is the fact that in vitro maturation is required post-thaw. The majority of live births from oocyte cryopreservation have involved the use of 1,2-propanediol and slow freezing protocols. Various methods have been used in an attempt to improve survival rates. These include vitrification and use of novel cryopreservatives. Future areas of concentration should include in vitro maturation, vitrification, and alternate cryopreservatives. PMID- 11941535 TI - Experimental models for ovarian tissue and immature follicles. AB - Primordial and growing follicles are abundant within the ovaries of healthy young female mammals. While our understanding of follicular dynamics is based mainly on studies of normal ovaries in intact animals, techniques such as ovarian grafting and in vitro culture, in particular when used in combination with cryopreservation, have provided both significant additional insights and a source of mature oocytes. Primordial follicles are small, quiescent, and most commonly located within the collagen-rich outer portion of the ovary. Providing that appropriate collection, handling, freezing, and thawing methods are used, they can tolerate cryopreservation very well irrespective of whether they are frozen within a whole ovary (small species), as ovarian pieces, or as individual, isolated, follicles. Animal studies show that grafts of such fresh or frozen materials can, providing that they contain viable follicles, form mature fertilizable oocytes, produce hormones, and support pregnancies to term. Grafts can be returned to the original donor (autograft), but grafting between histocompatible individuals of the same species and between species (xenografts) is also possible. Cryopreservation and grafting are therefore useful both as practical and as experimental tools. Clinically, ovarian tissue has started to be collected and frozen for patients who are at risk of ovarian failure. Very recent case reports show that such frozen ovarian tissue autografts can support the return of menses and antral follicle formation in patients, although as yet no pregnancies have been established. PMID- 11941536 TI - Preliminary experience with orthotopic and heterotopic transplantation of ovarian cortical strips. AB - The idea of fresh orthotopic autologous ovarian transplantation in humans is neither novel nor sophisticated; a New York surgeon reported on this technique as early as in 1906. It is the recent possibility of cryostorage of ovarian tissue and the development of new orthotopic and heterotopic techniques to autotransplant frozen-thawed ovarian cortical strips that brought a new dimension to the field. With the availability of more effective cryoprotectants, researchers were able to demonstrate that ovarian function and fertility could be restored after transplantation of cryopreserved ovarian tissue in large mammals. Experimental models of ovarian transplantation are discussed in the article by Shaw and Trounson; our focus will be on the clinical applications of ovarian transplantation and new developments in heterotopic transplantation. Patient selection and screening, details of the surgical techniques, and safety measures to avoid reseeding cancer cells via transplanted tissue will be discussed. Synchronization between the laboratory and the operating room will be detailed to provide guidance for clinicians who are contemplating ovarian transplantation with previously frozen ovarian tissue. PMID- 11941537 TI - Cryobiology ethics of human reproduction. AB - The article surveys the ethical and social issues that arise from recent developments in assisted reproductive technologies (ART) involving cryotechnology and argues that practitioners must understand ethical as well as technological issues. It discusses the risks associated with ART and the importance of the need to inform patients of them so that they can make considered decisions. It considers problematic areas in the use of ART, particularly the case of lesbian women who wish to conceive. It offers a philosophical definition of ethics and a framework for defining ethical, and unethical, clinical practice. It advocates a utilitarian approach to moral problems, one that attends to the moral consequences of actions. It examines the complex way that morality, law, and ethics interact and refers to controversial legal decisions in the United Kingdom concerning ART. This article considers the various rights (of donors, prospective parents, and offspring) that need to be taken into account by practitioners of ART. PMID- 11941538 TI - Structural and functional mutations of the perlecan gene cause Schwartz-Jampel syndrome, with myotonic myopathy and chondrodysplasia. AB - Perlecan, a large heparan sulfate proteoglycan, is a component of the basement membrane and other extracellular matrices and has been implicated in multiple biological functions. Mutations in the perlecan gene (HSPG2) cause two classes of skeletal disorders: the relatively mild Schwartz-Jampel syndrome (SJS) and severe neonatal lethal dyssegmental dysplasia, Silverman-Handmaker type (DDSH). SJS is an autosomal recessive skeletal dysplasia characterized by varying degrees of myotonia and chondrodysplasia, and patients with SJS survive. The molecular mechanism underlying the chondrodystrophic myotonia phenotype of SJS is unknown. In the present report, we identify five different mutations that resulted in various forms of perlecan in three unrelated patients with SJS. Heterozygous mutations in two patients with SJS either produced truncated perlecan that lacked domain V or significantly reduced levels of wild-type perlecan. The third patient had a homozygous 7-kb deletion that resulted in reduced amounts of nearly full length perlecan. Unlike DDSH, the SJS mutations result in different forms of perlecan in reduced levels that are secreted to the extracellular matrix and are likely partially functional. These findings suggest that perlecan has an important role in neuromuscular function and cartilage formation, and they define the molecular basis involved in the difference in the phenotypic severity between DDSH and SJS. PMID- 11941539 TI - Germline alterations of the RNASEL gene, a candidate HPC1 gene at 1q25, in patients and families with prostate cancer. AB - The RNASEL gene (2',5'-oligoisoadenylate-synthetase dependent) encodes a ribonuclease that mediates the antiviral and apoptotic activities of interferons. The RNASEL gene maps to the hereditary-prostate-cancer (HPC)-predisposition locus at 1q24-q25 (HPC1) and was recently shown to harbor truncating mutations in two families with linkage to HPC1. Here, we screened for RNASEL germline mutations in 66 Finnish patients with HPC, and we determined the frequency of the changes in the index patients from 116 families with HPC, in 492 patients with unselected prostate cancer (PRCA), in 223 patients with benign prostatic hyperplasia (BPH), and in 566 controls. A truncating mutation, E265X, was found in 5 (4.3%) of the 116 patients from families with HPC. This was significantly higher (odds ratio [OR] =4.56; P=.04) than the frequency of E265X in controls (1.8%). The highest mutation frequency (9.5%) was found in patients from families with four or more affected members. Possible segregation was detected only in a single family. However, the median age at disease onset for E265X carriers was 11 years less than that for noncarriers in the same families. In addition, of the four missense variants found, R462Q showed an association with HPC (OR=1.96; P=.07). None of the variants showed any differences between controls and either patients with BPH or patients with PRCA. We conclude that, although RNASEL mutations do not explain disease segregation in Finnish families with HPC, the variants are enriched in families with HPC that include more than two affected members and may also be associated with the age at disease onset. This suggests a possible modifying role in cancer predisposition. The impact that the RNASEL sequence variants have on PRCA burden at the population level seems small but deserves further study. PMID- 11941540 TI - Krit1 missense mutations lead to splicing errors in cerebral cavernous malformation. AB - At least 40% of families affected with cerebral cavernous malformation have a mutation in Krit1. We previously identified two point mutations in Krit1 leading to changes in amino acids (D137G and Q210E) in two different families. Further RNA analysis reveals that both point mutations actually activate cryptic splice donor sites, causing aberrant splicing and leading to a frameshift and protein truncation. To date, no simple missense mutations have been detected in Krit1. PMID- 11941541 TI - Empiric use of trimethoprim-sulfamethoxazole (TMP-SMX) in the treatment of women with uncomplicated urinary tract infections, in a geographical area with a high prevalence of TMP-SMX-resistant uropathogens. AB - This study evaluated whether trimethoprim-sulfamethoxazole (TMP-SMX) is effective for treatment of uncomplicated urinary tract infections (UTIs) due to TMP-SMX resistant (TMP-SMX-R) pathogens. Healthy nonpregnant premenopausal women with symptomatic lower UTI were assessed for the presence of pyuria and bacteriuria; if either was present, a urine sample was cultured and TMP-SMX was prescribed. Clinical and microbiologic cure was assessed at days 5-9 and 28-42 after cessation of therapy. For 71%, of patients, cultures grew TMP-SMX-susceptible (TMP-SMX-S) microorganisms, and for 29%, cultures grew TMP-SMX-R organisms. Escherichia coli remained the predominant bacteria in both groups of cultures. At visit 2, microbiological cure had been achieved in 86% of the patients in the TMP SMX-S group and 42% of those in the TMP-SMX-R group. Similar differences were found at visit 3 by clinical evaluation. Treatment with TMP-SMX of uncomplicated UTI caused by TMP-SMX-R microorganisms results in microbiologic and clinical failure. In high-resistance areas, TMP-SMX should not be the empiric drug of choice for uncomplicated UTI. PMID- 11941542 TI - Epidemiology of acute viral gastroenteritis in children hospitalized in Rouen, France. AB - This study assessed the epidemiologic characteristics of acute viral gastroenteritis in hospitalized children. A stool sample obtained from each child was analyzed for the presence of astrovirus, calicivirus, rotavirus, adenovirus, enterovirus, and digestive bacteria. Of the 438 stool samples obtained, 138 tested positive for > or =1 pathogen during the winters of 1997-1998 and 1998 1999 (P<.001). Virologic tests revealed rotavirus in 17.3% of samples, calicivirus in 7.3%, astrovirus in 6.8%, adenovirus in 0.7%, and > or =1 virus in 5.4%. Median age was higher for patients with rotavirus gastroenteritis than it was for those with astrovirus or calicivirus gastroenteritis (P=.014). Mean duration of hospitalization was statistically significantly lower for children with rotavirus gastroenteritis (P=.022), despite the more-frequent dehydration observed among children with rotavirus versus those with astrovirus or calicivirus gastroenteritis (P=.007). In contrast, enteral rehydration was more rapidly achieved in patients with gastroenteritis due to rotavirus. PMID- 11941543 TI - Venous thrombosis associated with peripherally inserted central catheters: a retrospective analysis of the Cleveland Clinic experience. AB - Peripherally inserted central catheters (PICCs) have become popular for long courses of intravenously administration of antibiotics. Although these devices are generally regarded as safe, thrombotic complications have been associated with their use. In a retrospective review, 51 (2.47%) of 2063 patients who had a PICC placed during 1994-1996 were found to have developed a total of 52 PICC associated venous thromboses (VTs). Two patients received the diagnosis of pulmonary embolism that was a complication of VT. Risk factors for VT identified by multiple logistic regression analysis were younger age, history of VT, discharge to a skilled-nursing facility, and therapy with amphotericin B. VT is a significant complication of PICC placement. It may occur more frequently than previously recognized and may be complicated by pulmonary embolism. Clinicians should maintain a high index of suspicion, especially for high-risk patients. PMID- 11941544 TI - Disease-specific diagnosis of coinfecting tickborne zoonoses: babesiosis, human granulocytic ehrlichiosis, and Lyme disease. AB - To determine whether a unique group of clinical and laboratory manifestations characterize certain major deer tick-transmitted human pathogens in North America, we compared the symptoms, short-term complications, and laboratory test results of New England residents who became ill due to > or =1 of these pathogens. Patients completed a uniformly structured questionnaire and submitted blood samples for serologic and polymerase chain reaction (PCR) testing after developing symptoms of Lyme disease, human babesiosis, or human granulocytic ehrlichiosis (HGE). Complete blood count with thin blood smear, PCR, and immunoglobulin M antibody tests helped differentiate the acute manifestations of these diseases. Physicians should consider use of tests designed to diagnose babesiosis and HGE in patients with Lyme disease who experience a prolonged flulike illness that fails to respond to appropriate antiborrelial therapy. PMID- 11941545 TI - Exchange transfusion as an adjunct therapy in severe Plasmodium falciparum malaria: a meta-analysis. AB - The efficacy of exchange transfusion as an adjunct treatment for severe falciparum malaria is controversial. No sufficiently powered, randomized, controlled study has been reported. We analyzed 8 studies that compared survival rates associated with adjunct exchange transfusion with those associated with antimalarial chemotherapy alone. Exchange transfusion was not associated with a higher survival rate than was antimalarial chemotherapy alone (odds ratio [OR], 1.2; 95% confidence interval [CI], 0.7-2.1). However, patients who received transfusions had higher levels of parasitemia and more-severe malaria. Sensitivity analysis found that survival rates were higher among patients with partial immunity to malaria (OR, 0.5; 95% CI, 0.2-1.2) than they were among patients with no immunity (OR, 2.1; 95% CI, 0.9-4.8; P=.007). Exchange transfusion does not appear to increase the survival rate; however, there were significant problems with the comparability of treatment groups in the studies reviewed, and a randomized controlled trial is necessary to determine whether exchange transfusion is beneficial. PMID- 11941546 TI - Parainfectious encephalomyeloradiculitis associated with herpes simplex virus 1 DNA in cerebrospinal fluid. AB - We describe a patient with acute encephalomyeloradiculitis associated with herpes simplex virus 1 (HSV-1) DNA in the cerebrospinal fluid (CSF), and we also review 4 similar cases previously reported from Japan. A 59-year-old man presented with acute encephalitis and urinary retention. Initially, coma and CSF pleocytosis improved with acyclovir treatment, but brain stem encephalitis, transverse myelitis, and lumbosacral polyradiculitis subsequently occurred. These conditions responded to corticosteroid therapy and immunoadsorption plasmapheresis. Polymerase chain reaction detected HSV-1 DNA in the CSF during acute encephalitis but not thereafter. Serial magnetic resonance imaging revealed transient lesions in the thalamus and basal ganglia on both sides of the brain and in the pons, spinal cord, and cauda equina. Acute encephalomyeloradiculitis is a unique neurological syndrome that may be caused by HSV-1 infection of the central nervous system. PMID- 11941547 TI - Gastrointestinal and hepatic manifestations of tickborne diseases in the United States. AB - Signs and symptoms related to the gastrointestinal tract and liver may provide important clues for the diagnosis of various tickborne diseases prevalent in different geographic areas of the United States. We review clinical and laboratory features that may be helpful in detecting a tickborne infection. Physicians evaluating patients who live in or travel to areas where tickborne diseases are endemic and who present with an acute febrile illness and gastrointestinal manifestations should maintain a high index of suspicion for one of these disease entities, particularly if the patient has received a tick bite. If detected early, many of these potentially serious illnesses can be easily and effectively treated, thereby avoiding serious morbidity and even death. PMID- 11941548 TI - Infectious complications of dental and periodontal diseases in the elderly population. AB - Retention of teeth into advanced age makes caries and periodontitis lifelong concerns. Dental caries occurs when acidic metabolites of oral streptococci dissolve enamel and dentin. Dissolution progresses to cavitation and, if untreated, to bacterial invasion of dental pulp, whereby oral bacteria access the bloodstream. Oral organisms have been linked to infections of the endocardium, meninges, mediastinum, vertebrae, hepatobiliary system, and prosthetic joints. Periodontitis is a pathogen-specific, lytic inflammatory reaction to dental plaque that degrades the tooth attachment. Periodontal disease is more severe and less readily controlled in people with diabetes; impaired glycemic control may exacerbate host response. Aspiration of oropharyngeal (including periodontal) pathogens is the dominant cause of nursing home-acquired pneumonia; factors reflecting poor oral health strongly correlate with increased risk of developing aspiration pneumonia. Bloodborne periodontopathic organisms may play a role in atherosclerosis. Daily oral hygiene practice and receipt of regular dental care are cost-effective means for minimizing morbidity of oral infections and their nonoral sequelae. PMID- 11941549 TI - Hantavirus pulmonary syndrome: the new American hemorrhagic fever. AB - The recognition of hantavirus pulmonary syndrome (HPS) after the investigation of a cluster of unexplained respiratory deaths in the southwestern United States during the spring of 1993 showcased our ability to recognize new and emerging diseases, given the correct juxtaposition of a new clinical entity with circumscribed epidemiologic features that are analyzed with novel diagnostic methods. In less than a decade, HPS has become established as a pan-American zoonosis due to numerous viruses maintained by sigmodontine rodents with rodent- and virus-specific epidemiologic profiles. The classical features of the syndrome acute febrile illness associated with prominent cardiorespiratory compromise after direct contact or inhalation of aerosolized rodent excreta-has been extended to include clinical variants, including disease with frank hemorrhage, that have confirmed that this syndrome is a viral hemorrhagic fever. Efforts are under way to refine prevention strategies, to understand the pathogenesis of the shock, and to identify therapeutic modalities. PMID- 11941550 TI - The promise of novel technology for the prevention of intravascular device related bloodstream infection. I. Pathogenesis and short-term devices. AB - Intravascular devices (IVDs) are widely used for vascular access but are associated with substantial risk of development of IVD-related bloodstream infection (BSI). The development of novel technologies, which are based on an understanding of pathogenesis, promises a quantum reduction in IVD-related infections in an era of growing nursing shortages. Infections of short-term IVDs (that is, those in place <10 days), including peripheral venous catheters, noncuffed and nontunneled central venous catheters (CVCs), and arterial catheters, derive mainly from microorganisms colonizing the skin around the insertion site, which most often gain access extraluminally. More-effective cutaneous antiseptics, such as chlorhexidine, a chlorhexidine-impregnated sponge dressing, CVCs with an anti-infective coating, anti-infective CVC hubs, and novel needleless connectors, have all been shown to reduce the risk of IVD-related BSI in prospective randomized trials. The challenge for the future will be to identify new preventative technologies and to begin to adapt more widely those technologies already shown to be efficacious and cost-effective. PMID- 11941552 TI - Morbidity and mortality in South African gold miners: impact of untreated disease due to human immunodeficiency virus. AB - A cohort of 1792 human immunodeficiency virus (HIV)-positive and 2970 HIV negative South African miners was observed for 12 months starting in February 1998. All-cause hospitalizations and deaths were significantly associated with HIV infection (respective unadjusted incidence rate ratios, 2.9 and 9.2; respective 95% confidence intervals, 2.5-3.4 and 5.5-16.0). Tuberculosis (TB), bacterial pneumonia, cryptococcosis, and trauma were the major causes of admission for HIV-positive patients, whereas Pneumocystis carinii pneumonia was an uncommon cause (respective admission rates, 8.5, 6.9, 2.2, 6.0, and 0.53 admissions per 100 person-years). Enteritis, bronchitis, urinary tract infections, and soft-tissue infections were also significantly associated with HIV infection. Cryptococcosis caused 44% of deaths among HIV-positive patients. Trauma was the main hazard for HIV-negative men, causing 42% of admissions and 60% of deaths. A broad range of infectious conditions is significantly associated with HIV infection in South African miners. Identification and implementation of effective prophylactic regimens are urgently needed. PMID- 11941551 TI - Randomized phase II trial of atovaquone with pyrimethamine or sulfadiazine for treatment of toxoplasmic encephalitis in patients with acquired immunodeficiency syndrome: ACTG 237/ANRS 039 Study. AIDS Clinical Trials Group 237/Agence Nationale de Recherche sur le SIDA, Essai 039. AB - In this international, noncomparative, randomized phase II trial, we evaluated the effectiveness and tolerance of atovaquone suspension (1500 mg orally twice daily) plus either pyrimethamine (75 mg per day after a 200-mg loading dose) or sulfadiazine (1500 mg 4 times daily) as treatment for acute disease (for 6 weeks) and as maintenance therapy (for 42 weeks) for toxoplasmic encephalitis (TE) in patients infected with human immunodeficiency virus. Twenty-one (75%) of 28 patients receiving pyrimethamine (95% lower confidence interval [CI], 58%) and 9 (82%) of 11 patients receiving sulfadiazine (95% lower CI, 53%) responded to treatment for acute disease. Of 20 patients in the maintenance phase, only 1 experienced relapse. Eleven (28%) of 40 eligible patients discontinued treatment as a result of adverse events, 9 because of nausea and vomiting or intolerance of the taste of the atovaquone suspension. Although gastrointestinal side effects were frequent, atovaquone-containing regimens are otherwise well tolerated and safe and may be useful for patients intolerant of standard regimens for toxoplasmic encephalitis. PMID- 11941553 TI - Hepatotoxicity associated with antiretroviral therapy containing dual versus single protease inhibitors in individuals coinfected with hepatitis C virus and human immunodeficiency virus. AB - To determine the rates of patients coinfected with human immunodeficiency virus (HIV) and hepatitis C virus (HCV) who discontinued therapy as a result of protease inhibitor (PI)-related hepatotoxicity, a retrospective review was conducted. Baseline CD4 counts, plasma HIV RNA levels, and duration of therapy were comparable between single- and dual-PI-treated subjects and between subjects receiving ritonavir-containing therapy and those receiving ritonavir-sparing therapy. The proportions of patients with elevations in alanine aminotransferase level to > or =5 times the upper limit of normal (19% versus 26%) and hyperbilirubinemia (30% versus 38%) were similar between the dual-PI (n=27) and single-PI treatment groups (n=39), respectively. No difference in these characteristics was observed between ritonavir-containing (n=34) and ritonavir sparing (n=32) treatment arms. Rates of treatment discontinuation due to hepatotoxicity were similar for single-PI and dual-PI therapy and for ritonavir containing and ritonavir-sparing regimens. Dual-PI therapy and inclusion of ritonavir do not seem to increase the rates of hepatotoxicity in PI-treated, HIV HCV coinfected subjects. PMID- 11941554 TI - Assessing eosinophil count as a marker of immune activation among human immunodeficiency virus-infected persons in sub-Saharan Africa. AB - In 611 human immunodeficiency virus-infected persons who had not yet begun to receive antiretroviral therapy, we evaluated the linear association between absolute eosinophil count (as a surrogate for immune response to helminthic infection) and CD4+ T cell count, and between absolute eosinophil count and log virus load. Overall, no significant correlations were observed between eosinophil count and CD4+ T cell count, or between eosinophil count and log virus load. PMID- 11941555 TI - Outbreak of enteroviral infection in a pediatric hematology-oncology unit. AB - We review the clinical courses and outcomes of an outbreak of enteroviral infection that occurred in 5 children with acute lymphoblastic leukemia during a 2-month period in a hematology-oncology unit. Three patients presented with encephalitis and 2 with parotitis. Three of the 5 patients recovered uneventfully and 2 died, 1 of chronic encephalitis and 1 of acute brain stem encephalitis. PMID- 11941556 TI - Subtype-dependent response of hepatitis B virus during the early phase of lamivudine treatment. AB - We conducted a 12-month longitudinal investigation of the subtype-dependent response of hepatitis B virus (HBV) to lamivudine treatment in 43 consecutive patients with chronic hepatitis B. HBV subtype ayw appears to respond better to lamivudine monotherapy than does HBV subtype adw (P=.005). This might be the reason for the lower incidence of lamivudine-resistant strains observed in persons infected with HBV subtype ayw during follow-up. PMID- 11941558 TI - Tolerability of antimalaria drugs. PMID- 11941559 TI - Uveitis due to Leishmania major as part of HAART-induced immune restitution syndrome in a patient with AIDS. PMID- 11941560 TI - Strongyloides stercoralis eggs in a urethral smear after bone marrow transplantation. PMID- 11941561 TI - Why prior fluconazole use is associated with an increased risk of invasive mold infections in immunosuppressed hosts: an alternative hypothesis. PMID- 11941562 TI - Safety of Lactobacillus strains used as probiotic agents. PMID- 11941565 TI - Meta-analysis of the impact of HIV on the infectiousness of tuberculosis: methodological concerns. PMID- 11941567 TI - Prevalence of the HIV protease mutation N88S causing hypersensitivity to amprenavir. PMID- 11941568 TI - Metabolic acidosis in HIV-infected patients. PMID- 11941569 TI - Group a streptococcal meningitis in adults: report of 41 cases and a review of the literature. AB - Chart review of 41 adult patients with group A streptococcal (GAS) meningitis in The Netherlands revealed that this is a community-acquired disease and occurs mainly in patients with predisposing factors: of the 41 patients in this case series, 24 (60%) of 40 evaluable patients had otitis or sinusitis. Fever and neck stiffness were the most common clinical manifestations of disease, but, in addition, high rates of seizures (12 [32%] of 38 patients), focal neurological findings (13 [36%] of 36 patients), and hyponatremia (20 [58%] of 35 patients) were found. In contrast with data from the literature that describes 27 adult cases, we found that GAS meningitis is a fulminant disease with a mortality rate of 27% (10 of 37 patients), and that neurological sequelae occur in 36% (12 of 38) of surviving patients. PMID- 11941570 TI - Unsuspected Toxoplasma gondii empyema in a bone marrow transplant recipient. AB - Toxoplasma gondii is an opportunistic parasite that can cause severe disease in immunosuppressed individuals. We report a case of unsuspected T. gondii empyema in a bone marrow transplant recipient that was diagnosed by the visualization of numerous intracellular and extracellular tachyzoites in Giemsa- and Gram-stained smears. The patient was treated with pyrimethamine, sulfadiazine, clindamycin, and atovaquone, and she survived 110 days after diagnosis, despite having a large parasite burden. PMID- 11941571 TI - Leptospirosis, water sports, and chemoprophylaxis. AB - Recreational activities, such as water sports and adventure travel, are emerging as an important risk factor for leptospirosis, a potentially fatal zoonosis. We report the clinical course of 2 patients who acquired leptospirosis through participation in water sports. Physicians caring for patients who participate in adventure travel involving water sports should be familiar with the risk factors for and diagnosis, prevention, and treatment of leptospirosis. PMID- 11941572 TI - The Medical Event Reporting System for Transfusion Medicine: will it help get the right blood to the right patient? AB - The Medical Event Reporting System for Transfusion Medicine (MERS-TM) collects, classifies, and analyzes events that potentially could compromise the safety of transfused blood to facilitate system improvement. This system is designed to collect data on near misses as well as actual events. Near-miss events are a valuable source of data because they occur more frequently than, but share many characteristics and causes of, actual events. Further, although most current reporting efforts describe only what has occurred with little attention to what caused the event, MERS-TM includes a standardized method of causal analysis. The standardization provided by MERS allows users to compare their experience with that of other organizations, which speeds learning across the entire transfusion medicine community. Important features of the MERS-TM system are that it is able to capture threats, hazards, near misses, injuries, and deaths; characterizes failures and recoveries systematically; identifies and provides causal codes for the entire range of system defects including technical, organizational, cultural, and human factors; raises staff awareness about error management; is easily integrated with existing quality assurance programs; has a consistent and straightforward classification method; enables compliance with mandatory Food and Drug Administration reporting and accreditation requirements; has features to deal with a high volume of reports; supplies Web-based training, data entry, and analysis; and provides comparative benchmarks from comparable institutions. PMID- 11941573 TI - Mutation databases and other online sites as a resource for transfusion medicine: history and attributes. AB - Recent advances in molecular biology and technology have provided evidence, at a molecular level, for long-known observations that the human genome is not unique but is characterized by individual sequence variation. At the present time, documentation of genetic variation occurring in a large number of genes is increasing exponentially. The characterization of alleles that encode a variety of blood group antigens has been particularly fruitful for transfusion medicine. Phenotypic variation, as identified by the serologic study of blood group variants, is required to identify the presence of a variant allele. Many of the other alleles currently recorded have been selected and identified on the basis of inherited disease traits. New approaches document single nucleotide polymorphisms that occur throughout the genome and best show how the DNA sequence varies in the human population. The primary data dealing with variant alleles or more general genomic variation are scattered throughout the scientific literature and only within the last few years has information begun to be organized into databases. This article provides guidance on how to access those databases online as a source of information about genetic variation for purposes of molecular, clinical, and diagnostic medicine, research, and teaching. The attributes of the sites are described. A more detailed view of the database dealing specifically with alleles of genes encoding the blood group antigens includes a brief preliminary analysis of the molecular basis for observed polymorphisms. Other online sites that may be particularly useful to the transfusion medicine readership as well as a brief historical account are also presented. PMID- 11941574 TI - Blood donors and factors impacting the blood donation decision. AB - The aging of the US population and the evidence that only about 5% of individuals in the United States donate blood each year raise concerns about the assurance of an adequate, safe supply of blood in the future. Blood donation decision making has been investigated worldwide for decades to understand the process better to increase donation efficiency, safety, retention, collection numbers, and diversity of the donor pool. This review focuses on the characteristics of allogeneic blood donors, the motivational sources in donor decision making, and the research concepts and techniques used to examine these factors. Some historic studies considered pivotal, as well as more recent surveys, may not be pertinent to or representative of the current national donor pool. Interpretation of data related to donor characteristics should examine whether demographics mirror the donor pool to assist in targeted recruitment or if targeted recruitment actually leads to the reported demographics. Few recent studies of donor motivation have been published. Modern sources of positive and negative motivation are worth exploring through scientifically sound investigations involving representative cohorts using multifactorial approaches. Strategies that focus on retaining return donors and transforming first-time donors into repeaters would be beneficial. Investigations are needed also to assess research questions and to develop well-designed interventions to test hypotheses and to produce generalizable findings applicable to future donor decision making. PMID- 11941575 TI - The impact of babesiosis on transfusion medicine. AB - Babesiosis is an emerging zoonotic disease that has begun to have a noticeable impact on transfusion medicine. This is reflected in the growing medical literature on the topic. There has been no review to summarize the various ways in which babesiosis influences transfusion medicine. Babesiosis is the most frequently reported tick-borne pathogen to be transmitted by blood transmission. Until recently, it has been an underrecognized complication of blood transfusion. However, the increased use of blood products for an ever-increasing elderly and immunodeficient patient population has heightened awareness about this disease. Fortunately, the risk of acquiring a symptomatic infection through a blood transfusion is surprisingly low. Nevertheless, babesiosis should be included in the differential diagnosis of a febrile hemolytic illness in recipients of blood transfusions. Infected individuals who become chronic carriers and donate blood during asymptomatic periods pose the greatest risk to the blood supply. The exact risk that this parasite poses to our blood supply remains to be accurately assessed. Reported cases of transmission, to date, have involved only the transfusion of red blood cells (RBCs) and, more rarely, platelets. Transmission of these piroplasms through plasma alone has not been documented. Much of our understanding about this organism evoked host responses and its requirements for in vitro survival has come from studies on non-human vertebrates. These studies have shown that antigenic variation may play a significant role in the development of prolonged parasitemia, that complement-induced changes to erythrocytes are pivotal in facilitating protozoan entry into host RBCs, and that autoimmunity contributes to disease. Severe infections may require lifesaving exchange transfusions and even plasmapheresis. Controlled studies to clearly define specific indications, benefits, and objectives of this therapy are still needed. Despite the development of novel and improved diagnostic tests, these tests are not readily available for the mass screening of blood donors. Improved strategies to assess and prevent transfusion-associated babesiosis are required. Current measures cannot be relied on to identify infected donors with a high degree of sensitivity or to protect susceptible recipients from this parasite. PMID- 11941576 TI - Possible mechanisms of allogeneic blood transfusion-associated postoperative infection. AB - Four possible mechanisms have been reported to underlie the apparent association of allogeneic blood transfusion (ABT) with postoperative bacterial infection. These are (1) a transfusion-related immunomodulatory (TRIM) effect of ABT mediated by immunologically active allogeneic white blood cells (WBCs) that downregulate the recipient's immune function, thereby predisposing to infection; (2) a TRIM effect of ABT mediated by soluble biologic response modifiers released in a time-dependent manner from WBC granules or membranes into the supernatant fluid of red blood cells (RBCs) during storage; (3) a TRIM effect of ABT mediated by soluble HLA peptides that circulate in allogeneic plasma; and (4) a related non-TRIM effect of ABT, whereby ABT causes postoperative organ dysfunction that predisposes to infection. The available clinical studies examining the association of ABT with postoperative infection were not designed to specifically test 1 or more of these 4 hypotheses. Thus, it is difficult to make inferences from the published data about the specific mechanism(s) that may underlie the purported association of ABT with infection. To permit such inferences, future studies of the association of ABT with postoperative infection should consider 2 outcomes (ie, postoperative organ dysfunction in addition to postoperative infection), as well as 3 possible exposures (ie, allogeneic WBCs, soluble biologic response modifiers originating in WBC granules or membranes, and/or soluble HLA molecules circulating in allogeneic plasma). PMID- 11941577 TI - Donor lymphocyte infusion therapy. AB - Donor lymphocyte infusion (DLI), which is therapy typically used after stem cell transplant, is a relatively new therapeutic intervention. It has been used in a number of hematopoietic malignancies, for some solid tumor malignancies, for its viral modulation effects, and as a prophylactic approach in the prevention of recurrent disease. The efficacy of this therapy is most evident in patients who have chronic myelogenous leukemia. However, studies of other diseases have been less forthcoming in showing clear benefit. Even in patient groups in which disease remission has been attributed to DLI, the risks of the treatment, namely graft-versus-host disease, may invalidate its use. This article summarizes a number of key studies addressing its usefulness in a number of disease states, risks of therapy, and strategies to ameliorate those risks while preserving benefit. It also examines tools that have been developed to more accurately describe survival benefits attributable to DLI. PMID- 11941579 TI - [An allelotype study of human glioblastoma]. AB - OBJECTIVE: To reveal the molecular genetic mechanisms for the pathogenesis of glioblastoma (GBM) and determine which chromosomes or chromosomal regions may play a role in the pathogenesis of GBM or may harbor tumor suppressor genes (TSGs) associated GBM. METHODS: An allelotype study of 21 cases of GBM was performed by polymerase chain reaction and loss of heterozygosity (LOH) analysis. Three hundred and eighty-two microsatellite markers covering all 22 autosomes were used. The mean genetic distance between two flanking markers is about 10 cM. Fluorescent dye-labeled primers and Perkin Elmer 377 DNA Sequencer were applied. RESULTS: LOH was observed on all chromosomal arms examined in this study. The LOH frequencies of 10q, 10p, 13q, 17p and 9p were the highest (>50%), on which high LOH frequencies were detected at the regions resided by the known TSGs including PTEN, DMBT1, p16, p15, p53 and Rb. The following commonly deleted regions were detected: 9p22-23, 10p12.2-14, 10q21.3, 13q12.1-14.1, 13q14.3-31, 17p11.2-12, 17p13, 3q24-27, 11p12-13, 14q31-32.3, 14q21-24.1, 22q13.2-13.3, 4q35, 4q31.1 31.2, 6qtel, 6q16.3. CONCLUSION: This study demonstrated that the pathogenesis of GBM is very complicated and associated with various molecular genetic abnormalities on lots of chromosomes. The chromosomal arms most closely relevant to the pathogenesis of GBM are 10q, 10p, 9p, 17p and 13q. Besides the well-known TSGs, such as PTEN, DMBT1, p16, p15, p53 and Rb, multiple unknown TSGs associated with GBM may be present on the commonly deleted regions observed for the first time in this study. PMID- 11941580 TI - [Molecular cloning of the human CX 58 gene]. AB - OBJECTIVE: To clone a novel human connexin gene and find out the relationship between this gene and hereditary deafness. METHODS: Through the basic local alignment search tool (BLAST) analysis against the database of expressed sequence tags (dbEST) of National Center for Biotechnology Information (NCBI) using the coding sequence of mouse Cx 57 gene, 9 novel ESTs were obtained and a contig was assembled. Nested polymerase chain reaction (PCR) and rapid amplification of cDNA ends (RACE) were performed using primers designed on the contig. A novel gene was obtained and was mapped by homologous analysis against human genome sequence. Mutation analysis was performed in 12 autosomal dominant hereditary deafness families. RESULTS: Nine ESTs were obtained by homologous analysis and a contig was assembled. Through nested PCR and RACE, a full length of cDNA was obtained from human liver, kidney Ready cDNA and placenta cDNA library, and was named CX 58. By comparison with human genome sequence, CX 58 was mapped at 1p32.3-p34.1. Mutation analysis of CX 58 was performed in 12 autosomal dominant hereditary deafness families, but no mutation was detected. CONCLUSION: A novel human connexin gene named CX 58 was cloned and mapped to 1p32.3-p34.1. The mutation of CX 58 may not result in autosomal dominant hereditary deafness. PMID- 11941582 TI - [Construction and expression of APP(SWE) transgenic mice model]. AB - OBJECTIVE: To prepare transgenic mice model of human Swedish mutation of amyloid precursor protein(APP(SWE)) gene. METHODS: By pronuclear microinjection, human APP (SWE) genes were microinjected into pronuclei of zygotes of C57 and Kunming white mice. Integral zygotes were implanted into the oviducts of pseudopregnant female mice. The integration and expression of exogeneous genes in offsprings were detected by polymerase chain reaction (PCR), fluorescence in situ hybridization (FISH) and reverse transcription-ploymerase chain reaction (RT-PCR) analysis. RESULTS: The success rate of microinjection and the birth rate were 76.62% and 10.38%, respectively. There were 6 founders and the integration rate was 35.29%. The exogeneous gene was transferred to the third generation. Expression of human APP(SWE) gene was detected in the brain, heart and muscle but not detected in the liver and kidney of transgenic mice. CONCLUSION: The transgenic mice model of human APP(SWE) gene has been prepared successfully. PMID- 11941581 TI - [Transmission disequilibrium test of DRD4 exon III 48bp variant number tandem repeat polymorphism and tic disorder]. AB - OBJECTIVE: To investigate whether DRD4 exon III48 bp variant number tandem repeat(VNTR) polymorphism is associated with tic disorder. METHODS: One hundred and twenty-two nucleus families were collected using Structured clinical interview for genetic study of Tourette syndrome and related disorders for family-based association analysis of tic disorder and DRD4 exon III 48bp VNTR polymorphism. One hundred and twenty-two trios were divided into two groups: tic disorder group (82 trios of Tourette syndrome or chronic tic disorder, TS&CT) and tic disorder accompanied with attention deficit and hyperactivity disorder (ADHD) group (40 trios of Tourette syndrome or chronic tic disorder accompanied with ADHD, TS&ADHD). Transmission disequilibrium test (TDT), in addition to polymerase chain reaction and VNTR technique were conducted in 122 trios. RESULTS: There exist 5 alleles at this polymorphic locus in this sample including DRD4 exon III 48bp 2-6 repeats. No transmission disequilibrium was found between DRD4 exon III 48 bp VNTR and tic disorder (chi square=7.44, P 0.12); however, when the sample was divided into two groups, transmission disequilibrium was noticed between the cases of TS&ADHD and this locus by overall allele-wise analysis (chi square=11.74, P 0.02), and there exists transmission disequilibrium exclusively between 5 or 6 repeats of 48bp VNTR(longer alleles) by allele-wise analysis (chi square=10.57, P 0.032, chi square=6.13, P 0.01). No transmission disequilibrium was seen between TS&CT and DRD4 exon III 48bp VNTR(chi square=3.38, P 0.50). CONCLUSION: The results of this study have revealed an association between the longer alleles of DRD4 exon III 48bp VNTR polymorphism and tic disorder accompanied with ADHD, thus suggesting a possible genetic risk factor of tic disorder accompanied with ADHD in Chinese. PMID- 11941583 TI - [Analyses of genetic model of psoriasis vulgaris]. AB - OBJECTIVE: To explore the possible genetic model of psoriasis vulgaris. METHODS: The complex segregation analysis and heritability calculation were performed with the aid of Penrose method, Falconer regression method and SAGE-REGTL program. RESULTS: It was found that in 1043 patients with psoriasis vulgaris, 305 patients (29.24%) have the family history of psoriasis, and 738 patients have not the family history. A ratio of s/q approached 1/(square root of q) with Penrose method, and the heritability values of psoriasis in the first-degree and second degree relatives were 67.04%, 46.6% respectively. By complex segregation analysis, Mendelian, non-major-gene model and environment model were rejected for psoriasis. CONCLUSION: The results suggest that psoriasis follows a pattern of polygenetic or multifactorial inheritance rather than single-gene inheritance. PMID- 11941584 TI - [Heritability of serum leptin levels: a twin study]. AB - OBJECTIVE: To study the contribution of genetic factors to the variance of serum leptin concentration in healthy, normotensive twins. METHODS: A total of 57 pairs of twins were investigated: 28 female and 19 male pairs of monozygotic(MZ) twins, and 6 female and 4 male pairs of dizygotic(DZ) twins. The zygosity of twins was determined by comparing the concordance of the genotype of nine fluorescence labeled microsatellite markers. The genetic analysis was performed using the variance-based method. Serum leptin levels were determined in duplicate by a radioimmunoassay Kit (Linco Research, Inc., St. Charles, Missouri) as previously described. RESULTS: The test of genetic variance revealed a significantly larger within-pair variance of serum leptin in the DZ twins, in comparison with the MZ twins. The corresponding heritability for serum leptin was 8%. Adjusted for BMI, gender, and uric acid (UA), the heritability for serum leptin was 0.18%. Log leptin correlated significantly with blood pressure (SBp r=0.355 P<0.001; DBp r=0.339 P<0.001). Stepwise multiple linear regression analysis revealed that only BMI, gender and UA were linked independently to serum leptin levels(R(2)=0.788, P<0.001). CONCLUSION: The above data indicate that environmental factors other than genetic factors are important determinants of leptinemia in normal subjects. PMID- 11941585 TI - [Variations of apolipoprotein A IV gene in Chinese endogenous hypertriglyceridemics]. AB - OBJECTIVE: The aim of this study was to investigate variations of apolipoprotein A IV (apo A IV) gene and its relation to endogenous hypertriglyceridemia(HTG) in Chinese population. METHODS: One hundred and six endogenous hypertriglyceridemics and 171 healthy subjects from a population of Chinese Han nationality in Chengdu area were studied using restriction fragment length polymorphisms (RFLPs) and sequencing of apoA IV gene amplified by polymerase chain reaction (PCR). The polymorphic sites of apo A IV gene studied included codon 9 (A to G, synonymous mutation), codon 347 (A to T, non-synonymous mutation), codon 360 (G to T, non synonymous mutation), and Msp I polymorphism (CC/TGG) within intron 2. RESULTS: The frequency of G allele at codon 9 in HTG group was higher than that in healthy controls(0.453 vs 0.366, P<0.05). The other polymorphic sites showed no significant differences of the allele frequencies between the two groups. The frequencies of rare alleles, such as G allele at codon 9, T allele at codon 347 and T allele at codon 360 polymorphic site were significantly different from those reported in European Caucasians (0.366 vs 0.032, P<0.001, 0.000 vs 0.160, P<0.001; 0.000 vs 0.070, P<0.001), but no differences were found when compared with those in Japanese, including Msp I site (P>0.05). In the healthy male control group, subjects with genotype G/G of codon 9 had a higher serum mean concentration of apoA I when compared with that of genotype A/A(P<0.01). In the HTG group, subjects with genotype C/T of Msp I site had a higher serum mean concentration of TG with compared with those with genotype C/C and T/T (P<0.05). This difference was only observed in male HTG group when male and female subgroups were further separated. CONCLUSION: These results suggest that Msp I and codon 9 polymorphism in apoA IV gene are associated with endogenous hypertriglyceridemia to some extent in Chinese population. PMID- 11941586 TI - [Study on the loss of heterozygosity and expression of transglutaminase 3 gene in laryngeal carcinoma]. AB - OBJECTIVE: To investigate the role of transglutaminase 3 (TGM3) gene in laryngeal carcinogenesis. METHODS: The authors detected the deletion indirectly through loss of heterozygosity (LOH) analysis at DNA level using 4 STR primers within and near TGM3 gene in 72 cases, and detected the differential expression of TGM3 gene in 8 cases of paired normal and cancerous tissue of laryngeal carcinoma by Northern blot. RESULTS: LOH was found existing in all of the microsatellite loci, and the LOH frequencies were 25.76%, 20.00%, 38.10% and 18.75% at D20S17, D20S607, D20S99 and D20S841 respectively; LOH concerning at least one polymorphism locus accounted for 61.11%. No correlation of clinical stage, lymph node metastasis and differentiation with the LOH of TGM3 gene was observed, P>0.05. TGM3 gene expressed significantly higher in normal tissues than in paired cancerous tissues. CONCLUSION: TGM3 gene might play an important role in laryngeal carcinogenesis and further researches will be needed to clarify the possible mechanisms. PMID- 11941587 TI - [Distribution of poly(ADP-ribose) polymerase pseudogene polymorphism and association with susceptibility to lung cancer in Chinese people]. AB - OBJECTIVE: To study the distribution of poly(ADP-ribose) polymerase(PARP) pseudogene polymorphism and the association with susceptibility to lung cancer in Chinese people. METHODS: The subjects of this study included 63 patients with lung cancer and 82 healthy controls matched in gender and age. Genome DNA was extracted from white blood cells. Products from PCR with a pair of specific primer were electrophoresized in agarose including EB. Under ultraviolet, observation and imaging were performed. RESULTS: There was no significant difference in genotype between the cases and controls. The frequencies of B allele in cases and controls were 0.095 and 0.116 respectively. Whether there was B allele or not, smoking was a risk factor of lung cancer (P<0.05). As the genotype was AA and AB or BB, smoking OR was 2.28 and 4.83 respectively. Among non-smokers, the risk at lung cancer did not increase in AB or BB genotypes(P=0.202). CONCLUSION: Frequency of B allele is relatively lower in Chinese people than in other races. In smokers, B allele may be a susceptible marker of lung cancer, and there is synergistic function between B allele and smoking. PMID- 11941588 TI - [Two single nucleotide polymorphisms of beta 2-adrenoceptor gene in elderly patients with hypertension]. AB - OBJECTIVE: To investigate the association of two single nucleotide polymorphisms (SNPs) of beta 2-adrenoceptor (beta 2-AR) gene with hypertension in elderly patients. METHODS: The study samples were collected from unrelated Chinese Han population of Dabie Mountain in Anhui province. Eighty-six elderly patients with hypertension and 43 controls were selected. Genotypes of +1053 and +1239 SNPs were typed by polymerase chain reaction-restriction fragment length polymorphism. RESULTS: The frequencies of the two SNPs complied well with the Hardy-Weinberg equilibrium in normal group. The distribution of genotypes AA, GA,GG of the SNP at locus +1239 in moderate and severe hypertension group was significantly different from that in normal group (chi square=8.67, P<0.05). There were evident differences in the frequencies of alleles of the two groups (chi square=4.02, P<0.05). No significant difference was observed in the distribution of genotypes of the SNP at locus +1053 between the two groups. CONCLUSION: These data indicate that the SNP at locus +1239 of beta 2-AR gene is associated with hypertension in elderly patients. PMID- 11941590 TI - [Study on genetic epidemiology of nasopharyngeal carcinoma in Guangdong, China]. AB - OBJECTIVE: To explore the characteristic of genetic epidemiology of nasopharyngeal carcinoma (NPC) in a high risk area Guangdong province, China. METHODS: Population investigation was made on the nuclear pedigrees of the first patient with NPC and his/her spouse, and then complex segregation analysis was performed using regressive Logistic model. RESULTS: The risk of suffering from NPC is 9.31 times higher in the first degree relatives of patient with NPC than in the first degree relatives of spouse. The separation ratio and heritability are 0.0588 (0.0182, 0.0994) and 68.08% respectively. The result of complex segregation analysis shows that model D is better than model A. CONCLUSION: The genetic trend and familial clustering of NPC are more significant and powerful in Guangdong. The risk of suffering from NPC is related with parent's state and senior sibling's state. Nasopharyngeal carcinoma is a multi-gene hereditary disease, but a single gene that decides the susceptibility to NPC may be present. PMID- 11941589 TI - [In vivo expression of human tissue-plasminogen activator transfected AGZY83-a cells implanted in mice]. AB - OBJECTIVE: To detect the expressing levels of human tissue-plasminogen activator(t-PA) in AGZY83-a cells transfected with pcDNA3.1(+) t-PA in vivo and the feasibility of using transplantation of cells for gene therapy of thrombotic diseases. METHODS: Expression vectors containing the t-PA cDNA gene were transfected into AGZY83-a cells. The transfected AGZY83-a cells were implanted into mice in different regions, and the plasma levels of human t-PA were assayed at intervals. RESULTS: The plasma levels of human t-PA were significantly increased in mice after implantation of transfected AGZY83-a cells and were significantly higher than those of control groups implanted with untransfected AGZY83-a cells. This significant increase lasted at least 105 days. The intraperitoneal implantation group expressed the highest level of human t-PA, a little higher than that of the subcutis implantation group, while both are much higher than that of the quadriceps femoris implantation group which expressed the lowest. CONCLUSION: The implanted transfected AGZY83-a cells are able to stably express high levels of human t-PA, and transplantation of cells transfected with pcDNA3.1(+) t-PA is a new promising method for gene therapy of thrombotic diseases. PMID- 11941591 TI - [Polymorphism of mitochondrial DNA region V in Bouyei people and Miao people living in Guizhou province of China]. AB - OBJECTIVE: To investigate the polymorphism of mitochondrial DNA region V in Bouyei people and Miao people living in Guizhou province of China. METHODS: Polymerase chain reaction and direct sequencing were used in the study. RESULTS: Only two kinds of polymorphism were found in Bouyei and Miao people. One was standard pattern, the other short pattern. And the frequencies of short pattern(9 bp deletion) in Bouyei and Miao people were 31.1% and 33.3% respectively. CONCLUSION: The polymorphisms of mitochondrial DNA region V in Bouyei people and Miao people living in Guizhou province of China are similar, but they are different from those in other people. PMID- 11941592 TI - [Analysis of three Y-STR loci polymorphism in isolated populations]. AB - OBJECTIVE: To figure out the polymorphism of three Y-STR loci in isolated populations and explore the consanguinity of the populations with the use of Y STR. METHODS: Male samples were selected from two isolated populations(80 and 60 males) in Zhejiang province and one open population (36 males), genescan was performed with males' DNA by genescan technology with ABI PRISM 377 sequencer at Y chromosome loci DYS388, DYS390 and DYS395. RESULTS: DYS388, DYS390, DYS395 allele counts in Yushan island population, Taohua island population and open population were 8, 9, 7, 5, 6, 7 and 6, 6, 5 respectively. Gene diversity was between 0.70-0.80 in the three populations. There was no difference in distribution of allele frequency and shared genotypes between the isolated populations and the open population by statistical test. Genetic distance is long between Taohua island population and open population, short between Yushan island population and open population, and moderate between Yushan island population and Taohua island population. CONCLUSION: The main allele is 129 at DYS388; 215 at DYS390; and 119 at DYS395. The distribution of allele frequency and gene diversity at DYS388, DYS390, DYS395 loci, and the shared genotypes between populations as well as the genetic distance are unable to explain the blood relationship between the isolated and open populations, suggesting the additional studies in large sample size will be necessary to use Y-STR for exploring the blood relationship between populations. PMID- 11941593 TI - [A simple and convenient method for construction of gene site-directed mutagenesis]. AB - OBJECTIVE: To introduce a new technique for rapid construction of gene site directed mutagenesis. METHODS: Three primers are synthesized. One is a primer with the needed mutation; the other two containing appropriate enzyme sites for construction of the PCR fragment into a suitable plasmid are located at the flanks of the mutation primer. After the amplification of the PCR fragment using the mutation primer and the reverse flanking primer, another PCR is performed using the previous PCR mutation segment as primer and the other flanking primer. The final PCR segment can be cloned into an appropriate plasmid by using the enzyme sites in the primers. RESULTS: Two site-directed mutagenesis have been successfully constructed in the Parkin gene by this method. CONCLUSION: The method is effective and simple for construction of gene site-directed mutagenesis. PMID- 11941594 TI - [Fluorescence probe two-hybridization technique and its application in medical genechips]. AB - OBJECTIVE: To establish a new nucleic acid hybridization detection technique which may be used in medical genechips. METHODS: The specific DNA fragment was detected by sequential two hybridization of fluorescence probe with template DNA and fixed DNA probe. RESULTS: Fluorescence probe two-hybridization (FPTH) was applied to genechips for the detection of sex-transmitted pathogens from culture strains, and the results showed that the values of fluorescence density of the positive groups decreased remarkably when compared with those of the negative group. Both the sensitivity and specificity for detecting clinical samples are higher than 90%. There is no need of any additional reagent in hybridization procedure, and the hybridization detection can be accomplished in 40 minutes. CONCLUSION: The FPTH technique is rapid, simple and reliable, it can also make the clinical detection process completely automatic and integrative. PMID- 11941595 TI - [Progress in molecular genetics of correlating genes of breast cancer]. AB - Good progress has been made in the researches on correlating genes of breast cancer in recent years. Quite a few kinds of genes such as susceptibility gene, oncogene and tumor suppressor genes have been found with implications for diagnosis, therapy and prognosis. Abnormality of breast cancer susceptibility gene (BRCA) is of great significance, especially in the development of breast cancer. PMID- 11941596 TI - [Pharmacogenomics genomics approaches to optimizing drug therapy]. AB - With the development of the research in human genomics, it is well known that genetic polymorphisms(mainly single nucleotide polymorphisms) of the genes encoding drug-metabolizing enzymes, transporters, receptors and other drug target proteins are relative to interindividual differences in the efficacy and toxicity of many medications. On the basis of functional genomics and molecular pharmacology, pharmacogenomics is elucidating the inherited nature of these differences in drug response mainly by means of analyzing the genetic variations of DNA and monitoring the gene expression pattern. It can not only improve disease diagnosis and predict the potential drug response but also speed up drug discovery and its development which will be instructive for clinical drug therapy ultimately. PMID- 11941597 TI - [Relationship between HFE gene and hereditary hemochromatosis]. AB - HFE gene is a major histocompatibility complex class I-like gene, which was identified as a candidate gene for hemochromatosis in 1996. The proposed role for HFE is its part in the regulation of the interaction of the transferrin receptor with transferrin. Hemochromatosis, the common autosomal recessive disease of iron overload, affects at least 1 in 300 Caucasians. The identification of the C282Y mutation in the HFE gene has led to population screening studies. Much of this work has also included the analysis of a second mutation, H63D, which appears to have a low penetrability. HFE protein was recently found to coprecipitate with the transferrin receptor and to affect the reaction between transferrin and the transferrin receptor. Functional data suggest that the mutation C282Y abolishes the association of the HFE protein with beta 2-microglobulin (beta 2M), making the complex unable to reach the cell surface. Clearly, if the mutation protein is unable to reach the cell surface, this regulatory feature is missing. The role of a second mutation in the HFE gene, H63D, is less clear. Current data suggest that this mutation protein can associate with beta 2-microglobulin and does reach the cell surface and that the defect lies in a failure to modify the affinity of the transferrin receptor for transferrin. This does not explain the low degree of penetrability associated with this mutation. PMID- 11941598 TI - [Surgical treatment of terminal heart failure]. AB - Terminal heart failure with no response to pharmacological treatment has been a challenge in medical practice: besides its deleterious effects on quality of life for all patients, it has a very high mortality rate. At present, surgical treatment of terminal heart failure consist basically in cardiac transplantation, which demands precise surgical indications, donor and receptor selection and adequate immunosuppression. Cardiac transplantation has contributed lower mortality rate and quality of life. Other surgical procedures that have been tried include cardiomyoplasty, partial left ventriculostomy (Batista's surgical procedure) and the use of several left ventricular mechanical devices. PMID- 11941599 TI - [Healing and destructive entanglements]. PMID- 11941600 TI - [Patient positioning before operation--care from A to Z]. PMID- 11941601 TI - [Dealing with elderly patients: multilevel aspects and fascinating]. PMID- 11941602 TI - [Political involvement of nurses. The Bern Hospital Movement]. PMID- 11941604 TI - [The good wishes of the queen]. PMID- 11941603 TI - [How learning can also be fun]. PMID- 11941605 TI - [Nursing care planning--a professional document needing revision]. PMID- 11941606 TI - [To each his own "normality"]. PMID- 11941607 TI - [Normal, normality and normative aspects: a reflection. "I am within the norm- you're not?"]. PMID- 11941608 TI - [Allowing a part of the ego to die as well]. PMID- 11941609 TI - [Nursing care on the floor. A simple method--surprising results]. PMID- 11941610 TI - [Geriatric rehabilitation. Indications for team communication]. PMID- 11941611 TI - [Psychiatry and social rupture. When the institution becomes a refuge]. PMID- 11941612 TI - [21 May 2000 vote. Bilateral agreement: consequences for patients]. PMID- 11941613 TI - [Supermom or super nurse?]. PMID- 11941618 TI - [Nursing care of AIDS patients. From fatal to chronic disease]. PMID- 11941619 TI - [Focus on pain. Into the depth of pain and beyond it]. PMID- 11941617 TI - [Empowerment for a profession]. PMID- 11941620 TI - [Those standing in the dark and living on the edge]. PMID- 11941621 TI - [Psychosocial health promotion. So that night becomes night time again]. PMID- 11941622 TI - [Never banal]. PMID- 11941623 TI - [Communicating in the hospital with the hearing impaired patient. "Madame B. doesn't understand"]. PMID- 11941625 TI - [Follow-up of patients with AIDS. The therapeutic pamphlet: a welcome aid]. PMID- 11941624 TI - [Care of persons with AIDS. After the crisis, chronicity]. PMID- 11941626 TI - ["Accountable for what we have initiated"]. PMID- 11941627 TI - ["Genetic manipulations--what effects on the development of man?"]. PMID- 11941630 TI - [Suffering]. PMID- 11941632 TI - [The opportunities of change]. PMID- 11941631 TI - [Aggression and violence in family caregiving]. PMID- 11941633 TI - [A course for refugee women. "Women give much in return"]. PMID- 11941634 TI - [A single smile]. PMID- 11941635 TI - [Encounter with Fiona's feet]. PMID- 11941639 TI - [The courage to respond to real questions]. PMID- 11941640 TI - [Accompanying and preparing children for hospitalization. Less fear, less drama, but smiles instead]. PMID- 11941641 TI - [The child, grief and school. Don't let silence take hold!]. PMID- 11941642 TI - [Strong women]. PMID- 11941643 TI - [Nursing care at the time of death. An intercultural approach]. PMID- 11941644 TI - [Alternation between theory and practice--results of a Fribourg survey. Study term, time of tension]. PMID- 11941645 TI - [3 approaches. Alternation: a concept, enlightenment]. PMID- 11941646 TI - [Caregiving.....due to sense of responsibility]. PMID- 11941647 TI - [Trehalose intolerance can be behind the abdominal symptoms caused by mushrooms]. PMID- 11941648 TI - [Colorectal cancer screening is profitable]. PMID- 11941649 TI - [An international epidemiologic comparison of allergies]. PMID- 11941650 TI - [Publication bias in the clinical drug research]. PMID- 11941652 TI - [The use of symptom--ECG registering in the evaluation of ambulatory arrhythmia patients]. PMID- 11941651 TI - [Congenital chloride diarrhea gene error in the anion transporter protein]. PMID- 11941653 TI - [Ureteropelvic rupture caused by a fall]. PMID- 11941654 TI - [Testis]. PMID- 11941655 TI - [Curse of the Byron family--is bipolar disease inheritable?]. PMID- 11941656 TI - [Bipolar disorder--a challenge for the adolescent psychiatry]. PMID- 11941657 TI - [Diagnosis and clinical course of bipolar disorder]. PMID- 11941658 TI - [Hypomanias and depressions--type II bipolar disorder]. PMID- 11941659 TI - [Has bipolar disorder been forgotten?]. PMID- 11941660 TI - [The changing drug therapy of bipolar disorder]. PMID- 11941661 TI - [Bipolar disorder and psychotherapies]. PMID- 11941662 TI - [Biological factors in bipolar disorder]. PMID- 11941663 TI - [Nursing data and abbreviation Chinese]. PMID- 11941664 TI - [Benefits for nurses and patients]. PMID- 11941665 TI - [Self control. Mrs. Katze's eyes begin to shine again]. PMID- 11941666 TI - [Anthroposophic medicine and nursing care. Holistic care at the center]. PMID- 11941668 TI - [Nursing care in movement]. PMID- 11941667 TI - [Between door and hook]. PMID- 11941669 TI - [Professionals at risk and stress. Dare to talk, cry and take time]. PMID- 11941671 TI - [New prospective]. PMID- 11941670 TI - [Unity of research and the development of patient care at CHUV. Missions touch all aspects of care]. PMID- 11941673 TI - [Better cards in hand with data]. PMID- 11941674 TI - [Healing poisoning arrows]. PMID- 11941675 TI - [Case analysis. Overcoming difficulties together]. PMID- 11941676 TI - [Crisis management. Attending to questions and anxiety of victims]. PMID- 11941677 TI - [Breaking away from role obscurity]. PMID- 11941678 TI - [Preventive home nursing visits. For independent living in old age]. PMID- 11941680 TI - [Women of the terrain]. PMID- 11941679 TI - [Innovative ideas for medicine and nursing care (interview by Urs Luthi)]. PMID- 11941681 TI - [Mediation. A means for appeasing the tempest]. PMID- 11941682 TI - [Autonomy of care. The end of the auxiliary status]. PMID- 11941683 TI - [Dental prevention in children of asylum seekers. Healthy teeth, thanks to the "smile" project]. PMID- 11941684 TI - [The role of writing in nursing care. The language of professionalism]. PMID- 11941685 TI - [Clinician specialist in diabetology in Nyon. "Crisis and chronicity--wanting to the answers" (interview by Francoise Taillens]. PMID- 11941688 TI - Educating future nurses. PMID- 11941689 TI - From grief to action: meeting the needs of bereaved families. PMID- 11941690 TI - STD epidemic reemerging? HPV most likely most common STD in adolescents. PMID- 11941691 TI - Success even in gridlock: exploring health policy victories in the 106th Congress. PMID- 11941692 TI - Return of the morning sickness pill. Could Bendectin be coming back to market? PMID- 11941693 TI - Breathe in breathe out. Controlling asthma during pregnancy. PMID- 11941694 TI - Preventing low birth weight. How smoking cessation counseling can help. PMID- 11941695 TI - Breastfeeding with multiple sclerosis. Helping women confront their fears. PMID- 11941696 TI - Breastfeeding promotion. Top tips for motivating women to breastfeed their infants. PMID- 11941698 TI - Counseling women with AIDS. Skills and tools every nurse needs. PMID- 11941699 TI - Curing & caring. PMID- 11941700 TI - Grief after abortion. PMID- 11941701 TI - Grief after abortion. PMID- 11941702 TI - Grief after abortion. PMID- 11941703 TI - [Illness-inducing myth]. PMID- 11941704 TI - [Postpartum exhaustion. When motherhood fatigues mothers]. PMID- 11941705 TI - [Nursing autonomy. No longer physician aids]. PMID- 11941706 TI - [Responsible for what we become familiar with]. PMID- 11941707 TI - [Violence toward women. What can health professionals do?]. PMID- 11941708 TI - [Exchange with Senegal instructors. Dynamics of formation and transformation]. PMID- 11941710 TI - [Intercultural nursing: encounter with the other]. PMID- 11941709 TI - [Diplomatic words no longer surfice]. PMID- 11941711 TI - [Incorporation of professionals. The formalities? Easy thanks to a memory aid]. PMID- 11941712 TI - [The smile of nurses under all circumstances]. PMID- 11941713 TI - [Communication in stress situations. Talking past each other can endanger health]. PMID- 11941714 TI - [Transactional analysis. Autonomy in communication]. PMID- 11941715 TI - [Autonomous profile for psychiatric nursing of the future]. PMID- 11941716 TI - [Ecology in the hospital. 5.4 kilogram waste per nursing care day]. PMID- 11941717 TI - [Assisted dying. Wandering on a "narrow edge"]. PMID- 11941718 TI - [Langnau regional hospital. Wraps and emollients on the nursing care plan]. PMID- 11941719 TI - [We are the artisans of our profession]. PMID- 11941720 TI - [Ethics of the subject. The body--at the heart of the nursing relationship]. PMID- 11941721 TI - [Negotiations with psychiatric patients. Of protocol by contract]. PMID- 11941723 TI - [Constructing one's future by standard curriculum]. PMID- 11941722 TI - [Preventing falls of patients in the hospital. Toward a comprehensive intervention program]. PMID- 11941724 TI - [Nurse of travel health. A specialty to promote]. PMID- 11941728 TI - [The nursing profession is our business]. PMID- 11941729 TI - [Nursing perception]. PMID- 11941730 TI - [Irchel Congress of SGGP. Building bridges]. PMID- 11941731 TI - [Older nurses. Actively engaged up to retirement]. PMID- 11941735 TI - [Unusual questions]. PMID- 11941736 TI - ["No blood": what should be done?]. PMID- 11941737 TI - [Appearances sometimes conceal a trauma]. PMID- 11941738 TI - [Clinical situation: when the life of a child is in danger. Ethical considerations in refusal of blood transfusion]. PMID- 11941739 TI - [Prevention, maintenance and promotion of help. Students plunge into "inside adventure"]]. PMID- 11941740 TI - [Transforming the dream of satisfactory professional writing into reality. Targeted transmissions]. PMID- 11941741 TI - [Conceptualizing nursing care. Use images]. PMID- 11941742 TI - [Needs in psychiatry. A relative notion]. PMID- 11941743 TI - [Humor in nursing care. Laughter frees life forces]. PMID- 11941744 TI - [6th Congress of the European Society of Palliative Nursing. Time stands still and races away]. PMID- 11941745 TI - [Death is a part of life]. PMID- 11941746 TI - [Kinesthetics. More than "pressure and pull"]. PMID- 11941748 TI - [Importance of home for man and the loss of it by entering a medico-social establishment. No more talk of terminal care]. PMID- 11941749 TI - [A kernel of hope]. PMID- 11941750 TI - [Adequate management of postoperative pain. References for the nursing team]. PMID- 11941751 TI - [Putting a face on violence. Finding words, choosing images]. PMID- 11941752 TI - [The Basel Humor Congress. What's so funny?]. PMID- 11941753 TI - [Political identity: luxury or necessity?]. PMID- 11941754 TI - [Effective, but not spectacular]. PMID- 11941755 TI - [Educational exchange. "Best experience of the entire educational process"]. PMID- 11941756 TI - [Great Britain. Massive money injection for nursing care]. PMID- 11941757 TI - [Personnel shortage in nursing care. Multilevel problems require multilevel solutions]. PMID- 11941758 TI - [Team work. Basics of a new organizational culture]. PMID- 11941759 TI - [The global budget. Hospitals develop into enterprises: advantages or disadvantages?]. PMID- 11941760 TI - [Imagine this]. PMID- 11941761 TI - [Creative nursing. The "difficult patient"]. PMID- 11941762 TI - [Era of the nursing team]. PMID- 11941763 TI - [Feeding patients in geriatric psychiatry. "You can manage a small spoonful, right?"]. PMID- 11941764 TI - [An innovative and structured concept. Management of episodes of care]. PMID- 11941765 TI - [Survey: Vaud nurses in team nursing care. The team member: an intensification without failing!]. PMID- 11941766 TI - [Wound consultation at the Loex Hospital. "Transmit knowledge above everything else"]. PMID- 11941769 TI - ["Dear Susan, nursing must change"]. PMID- 11941770 TI - [Quality assurance and efficiency are not contradictions (interview by Urs Luthi)]. PMID- 11941771 TI - [Needs assessment. 2 Spitex services compared]. PMID- 11941772 TI - [Interdisciplinary cooperation. Pain therapy at home lowers costs]. PMID- 11941773 TI - [The approach is important]. PMID- 11941774 TI - [Social aspects and nursing care. Communicating what we do]. PMID- 11941775 TI - [Social aspects and nursing care. We counsel ill human beings]. PMID- 11941776 TI - [Not an easy life]. PMID- 11941777 TI - [The proper balance]. PMID- 11941779 TI - [Human-animal relationship. A bond toward the end of life]. PMID- 11941778 TI - [Do you speak jackal or giraffe?]. PMID- 11941780 TI - [Learning to communicate in a non-violent manner. When conflict transforms into dialogue]. PMID- 11941781 TI - [Embracing change. Coaching in health care]. PMID- 11941782 TI - [International Year of the Culture of Peace. Against violence on all fronts]. PMID- 11941783 TI - [Health promotion. Toward a night is a true night]. PMID- 11941784 TI - [Modification of a nursing practice based on evidence. Hygienic washing and disinfection of hands]. PMID- 11941787 TI - [Spitex--focus of health care in the future]. PMID- 11941788 TI - [Active euthanasia: more transparency or rupture of the dike? (interview by Urs Luthi)]. PMID- 11941789 TI - [University study. Hospital directors distribute grades]. PMID- 11941790 TI - [Blood donation. New test for hepatitis C viruses]. PMID- 11941791 TI - [How much space does a person need?]. PMID- 11941794 TI - [Right to health]. PMID- 11941795 TI - [Why not a bit bolder?]. PMID- 11941796 TI - [Pedagogy and professional project]. PMID- 11941797 TI - [Cardiovascular rehabilitation and prevention. Specific training for nurses?]. PMID- 11941798 TI - [Is the vancomycin resistance of Staphylococcus aureus left unnoticed?]. PMID- 11941799 TI - [Day surgery is expanding--in its own unit or inside the operative department?]. PMID- 11941800 TI - [The present-day of ultrasonographic techniques]. PMID- 11941801 TI - [Protecting kidneys from radiocontrast toxicity--do fluids or drugs help?]. PMID- 11941802 TI - [Local treatment of pyoderma gangrenosum with cromoglycate solution]. PMID- 11941803 TI - [Ovary--a constantly changing organ]. PMID- 11941804 TI - [Treatment guidelines help to beat skin infections and respiratory infections]. PMID- 11941805 TI - [Treatment guidelines for primary bacterial skin infections]. PMID- 11941806 TI - [Recommended treatment of acute sinusitis]. PMID- 11941808 TI - [Recommended treatment of pharyngitis]. PMID- 11941807 TI - [Recommended treatment of acute otitis]. PMID- 11941809 TI - [Recommended treatment of acute bronchitis]. PMID- 11941810 TI - [Breast milk and allergies]. PMID- 11941811 TI - [Refractive surgery: the call of sirens or farewell to eyeglasses?]. PMID- 11941812 TI - [Mutation of the cytokine receptor behind periodical fever syndrome]. PMID- 11941813 TI - [Immunological aspects of human breast milk]. PMID- 11941814 TI - [Tumor necrosis factor modulators in the treatment of rheumatoid arthritis]. PMID- 11941815 TI - [Cerebrovascular disorders and the autonomic nervous system]. PMID- 11941816 TI - [Words, emotions and the health]. PMID- 11941817 TI - [Cleavage of corpus callosum in the treatment of severe epilepsy in children and adolescents]. PMID- 11941818 TI - [Difficult dysphagia in a young man--diabetes, alcohol or sarcoidosis?]. PMID- 11941819 TI - [Problem-based learning needed even in the further education]. PMID- 11941820 TI - [What else make you loose weight?]. PMID- 11941821 TI - [Is Bethesda system better than Pap classification?]. PMID- 11941822 TI - [Nursing is worth the study curriculum]. PMID- 11941823 TI - [So that the partner doesn't turn into a frog herself]. PMID- 11941824 TI - [Quality of life for patients]. PMID- 11941825 TI - [A question of attitude]. PMID- 11941828 TI - [An existence worthy of its name]. PMID- 11941829 TI - [Precariousness and costs of care. A mobile unit for the most deprived]. PMID- 11941830 TI - [Impoverishment of the multi-drug addicted in the Vaud canton. Adapted responses but insufficient]. PMID- 11941831 TI - [Education plays the key role]. PMID- 11941832 TI - [Care and security behind bars]. PMID- 11941833 TI - [A road between order and disorder]. PMID- 11941834 TI - [From nurse to public health expert]. PMID- 11941835 TI - [The soul also participates in the bath]. PMID- 11941836 TI - [Nursing care with many facets]. PMID- 11941837 TI - [ICN addresses requirements for the future]. PMID- 11941838 TI - [Hepatitis C. Alarming increase of drug dependent patients]. PMID- 11941841 TI - [The measure is full]. PMID- 11941842 TI - [The entire truth]. PMID- 11941843 TI - [Reinforcing the competence of nurses in Albania]. PMID- 11941844 TI - [Reflections on an experience in development. Interference or cooperation: the stakes of asymmetry]. PMID- 11941845 TI - [Nursing care quality is no longer guaranteed--SBK sounds the alarm]. PMID- 11941846 TI - [Prevention and health promotion in the community. "I rely on the links of a chain" (interview by Francoise Taillens]. PMID- 11941847 TI - [Study of 260 Swiss students. Personality and scholarly pursuits: profile of success]. PMID- 11941848 TI - [Patients and nurses. Trust--the "certain something"]. PMID- 11941849 TI - [50 years WE'G. "Achieving specialty identity is hard work"]. PMID- 11941850 TI - ["The graduate education market will promote us" (interview by Urs Luthi)]. PMID- 11941851 TI - [Stress and coping with stress. Feeling well, being healthy]. PMID- 11941852 TI - [22. Meeting of delegates. New leadership structure and contributing policies are on the table]. PMID- 11941853 TI - [What has helped?]. PMID- 11941854 TI - [Ethics in routine nursing practice. Students in an ethical conflict situation]. PMID- 11941855 TI - [The temptation of independence]. PMID- 11941856 TI - [Practicing as an independent nurse. "Take the chance, you will be recompensated"]. PMID- 11941857 TI - ["Demystifying the image image of mother of a family who keeps records by hand"]. PMID- 11941858 TI - [L'ADIIS. A young and dynamic association]. PMID- 11941859 TI - [European model of quality management. Independent nurses are also involved]. PMID- 11941860 TI - [Scarcity of nursing personnel. The problem is complex, but solutions exist]. PMID- 11941861 TI - [International Congress of La Source. Managing "with" instead of managing "for"]. PMID- 11941862 TI - [A question of trust]. PMID- 11941863 TI - [Ethics and progress]. PMID- 11941864 TI - [Concret AG nursing quality on the intensive care unit under evaluation]. PMID- 11941865 TI - [Family. Beside the dying they are often forgotten]. PMID- 11941866 TI - [Managing grief. Universally human and individual at the same time]. PMID- 11941867 TI - [Unexpected death. Why my child?]. PMID- 11941868 TI - [17th Research Congress of SBK. Evidence-based nursing care--a contribution to better quality of life]. PMID- 11941870 TI - [AIDS help in Switzerland. 3 new brochures about HIV/AIDS]. PMID- 11941869 TI - [Intuition]. PMID- 11941871 TI - [Chemotherapy--for the patient or to please the family?]. PMID- 11941872 TI - [And nevertheless, all changes...]. PMID- 11941873 TI - [Pilot study of nursing homes in the Fribourg canton. Maltreatment of acknowledgement]. PMID- 11941874 TI - [Creation of a personality disorders section. A new project]. PMID- 11941875 TI - [17th Research Meeting of ASI. Nursing practice founded on proof]. PMID- 11941876 TI - [Consultation for diabetics at Geneva University Hospital. End the separation by age!]. PMID- 11941877 TI - [Contribution of nursing care in society. "The human dimension is central" (interview by Margrit Bachl, Urs Luthi)]. PMID- 11941878 TI - [Anthroposophy in nursing care. Art of life, art of war]. PMID- 11941879 TI - [Nursing trials]. PMID- 11941880 TI - [Courage in the face of death]. PMID- 11941881 TI - [Basic principle determines the direction]. PMID- 11941882 TI - [Private conversations. Part of the process]. PMID- 11941884 TI - [Anesthesia nursing. Stress in emergency interventions]. PMID- 11941883 TI - [Nursing standard. Managing patients with chronic illnesses]. PMID- 11941885 TI - [Neglected]. PMID- 11941886 TI - [Career counseling. Thinking the taboo to a conclusion]. PMID- 11941887 TI - [Beyond the confusion of languages]. PMID- 11941888 TI - [Teaching diabetic refugees. Say it in Serbo-Croatian]. PMID- 11941889 TI - [Caring for immigrants. Understanding and comprehension, the fundamental right of the patient/client]. PMID- 11941890 TI - [The power to pull oneself out of the swamp by one's own hair]. PMID- 11941891 TI - [Prevention and control of infection. Training adapted to current realities]. PMID- 11941892 TI - [Planning nursing care. A work aspect which merits revisiting]. PMID- 11941894 TI - [SBK Congress 2000 in Montreux. Country-wide strike is not a taboo any more]. PMID- 11941895 TI - Dermatitis, glossitis, stomatitis, cheilitis, anemia and weight loss: a classic presentation of pancreatic glucagonoma. AB - Glucagonomas are rare tumors. They are predominantly located in the body or tail of the pancreas and display a constellation of signs and symptoms referred to as glucagonoma syndrome. The term necrolytic migratory erythema is used to characterize the distinctive rash associated with this syndrome. This report describes a classic presentation consisting of dermatitis, glossitis, stomatitis, angular cheilitis, anemia, and weight loss that was associated with the finding of a pancreatic mass and a markedly elevated plasma glucagon level. After pancreatic resection, the patient had complete resolution of the rash and normalization of plasma glucagon. PMID- 11941896 TI - Cephalic vein cutdown for inserting indwelling subclavian vein catheters in gynecologic oncology patients. AB - To decide on an alternate procedure for placement of central vein catheters other than percutaneous access of the subclavian vein, we conducted a study of 46 women who underwent placement of a central venous catheter utilizing the cephalic vein procedure at Ruby Memorial Hospital at West Virginia University in Morgantown. Of these patients, 96% had a successful placement of a catheter into the subclavian vein following dissection of the cephalic vein within the deltopectoral groove. There were no cases of pneumothorax or injury to the great vessels during the surgical procedure, which is a significant advantage for women who often have significant pleural effusions. PMID- 11941897 TI - Making the sale: communicating the importance of smoking cessation to pregnant patients. AB - The risks that smoking pose to the pregnant patient and her fetus have been well established. Some women quit smoking upon learning they are pregnant, but most do not. Although a direct, succinct intervention by the physician has proven to be a critical initial step in helping the patient to quit smoking, some physicians are not taking advantage of this opportunity. This article provides data obtained from focus groups conducted with pregnant West Virginia smokers. These women spoke very candidly about their experiences with their providers. Chances for succeeding with helping a patient to quit smoking are likely to increase if physicians discuss smoking with their patients at each visit and if it is done in a constructive manner. PMID- 11941899 TI - Genetic markers of hypertension: what will the future bring? PMID- 11941898 TI - Blood pressure and genetic and biological markers in a pediatric population. AB - BACKGROUND: It is universally accepted that atherosclerosis originates in childhood and that high LDL and low HDL serum cholesterol, hypertension and obesity are some of the main risk factors. In the past few years, increasing importance has been attributed to the deleterious effect of reactive oxygen species (ROS) on lipids, on the endothelial lining of arteries, and on the occurrence of cardiovascular disease. The relationship between blood pressure, active renin (AR) and the genetic polymorphism of haptoglobin in a population of adolescents was studied using pro-oxidant status markers, and also using some enzyme systems involved in the antioxidant defense mechanism of the body. METHODS: 51 healthy children, 9 to 12 years of age, were evaluated. The following examinations were performed: nutritional status (weight, height, skinfolds and body mass index) and blood pressure; note was made of any family history of cardiovascular disease. The biological parameters assessed by internationally recommended methods included haptoglobin phenotyping and determination of active renin (AR), the Na+/Li+ countertransport system (Na+/Li+ CTR), antioxidant enzyme systems, metahemoglobin reductase (MetHbRed) and transmembrane reductase (TMR) and indicators of oxidative status, namely activity of plasma epinephrine oxidase (EO) and erythrocytic acid phosphatase (LMW-PTP). RESULTS: The study of the behavior of AR, Na+/Li+ CTR, EO and erythrocyte enzymes (MetHbRed, TMR and LMW PTP) according to haptoglobin genetic phenotypes, only showed a significant difference for renin, with lower values for allele 1 carriers (homo- and heterozygous) (p < 0.05). There were no significant differences in blood pressure as regards the haptoglobin genetic phenotypes. The analysis of the behavior of the same biochemical parameters considering a family history of cardiovascular disease (CVD) showed higher values of TMR and lower values of EO in individuals with negative family history. Linear multiple regression showed some biological (Na+/Li+ CTR and MetHbRed) and genetic parameters (haptoglobin and family history of cardiovascular disease) to be stronger determinants of systolic blood pressure. CONCLUSIONS: It would appear that there is a tendency for some intermediate biochemical phenotypes to express early on in childhood in individuals with a family history of CVD, which would point to an evident genetic disposition towards this type of pathology. PMID- 11941900 TI - Perinatal arrhythmias -- diagnosis and treatment. AB - We did a retrospective analysis of the clinical files of 26 neonates with arrhythmia born during the period between January 1994 and February 1999. Fourteen (53.8%) of the neonates were male and 16 (61.5%) had prenatal diagnosis. Four (15.3%) had associated congenital heart disease. Twenty-one (80.7%) had abnormal rhythm and five (19.2%) had cardiac conduction disorder. Digoxin was the therapy of first choice to restore normal rhythm, used in 66.6% of the patients, followed by adenosine (16.6%) and electrical cardioversion (16.6%). An epicardial pacemaker was used in two of the three neonates with complete atrioventricular (AV) block. One neonate died due to cerebral hemorrhage. All the neonates were discharged in a clinically stable condition and 16 (88.8%) of them were medicated with digoxin. In a follow-up period that varied from 0 to 71 months (mean of 30.8 months), two patients had an episode of supraventricular tachycardia (SVT) after treatment withdrawal. Perinatal arrhythmias, although uncommon, can be life threatening, and hence we consider our experience with these situations worth presenting. PMID- 11941901 TI - [Ionized calcium and magnesium as interdependent variables in blood pressure origin in healthy adolescents]. AB - INTRODUCTION: Growing evidence indicates that diet divalent cations--ionized calcium (Ca) and magnesium (Mg)--and MN blood group polymorphism may be associated with essential arterial hypertension. OBJECTIVES: To assess a possible relationship between serum ionized Ca and Mg and MN blood group polymorphism with blood pressure (BP) and body mass index (BMI). POPULATION AND METHODS: The study group consisted of 173 healthy adolescents, 96 female (55.5%) and 77 male (44.5%), with mean age of 13.0 +/- 1.7 years. Serum ionized Ca and Mg were determined by standardized methods based on detection by specific electrodes. MN blood group polymorphism was determined by immunoaglutination of erythrocytes with monoclonal antibodies. BP was evaluated based on AHA recommendations. BMI was calculated as weight divided by squared height (Kg/m2). The statistical method used was the Pearson correlation. RESULTS: In this population no significant correlation was found from serum ionized Ca (mean: 1.07 +/- 0.06 mmol/L) and Mg (mean: 1.15 +/- 0.97 mmol/L) with systolic BP (mean: 113.1 +/- 10.7 mmHg) and diastolic BP (mean: 66.1 +/- 10.1 mmHg). In contrast there was an inverse correlation between the quotient ionized Mg/ionized Ca (0.45 +/- 0.05) and diastolic BP (p = 0.029; r = -0.381). A positive correlation was found between BMI (20.57 +/- 3.24 Kg/m2) and systolic BP (p = 0.007; r = 0.24) and diastolic BP (p = 0.016; r = 0.22). No correlation was found between ionized Ca, ionized Mg and BP with MN blood group polymorphism. CONCLUSIONS: In this population of healthy adolescents there was a dependence between diastolic BP and serum ionized Mg/ionized Ca. There was no interindividual BP and ionized Ca and Mg variation dependent from genetic variation. PMID- 11941902 TI - [Erythrocytic acid phosphatase genetic polymorphism and cardiovascular risk in health children and adolescents]. AB - OBJECTIVE: To correlate, in a sample of healthy children and adolescents, the activity of the enzyme acid phosphatase (ACP) with its different genetic phenotypes and of these with some cardiovascular risk parameters such as body mass index (BMI), percentage of total fat mass (%TFM), trunk fat (TF), insulin resistance, and the arterial blood pressure (BP). DESIGN AND METHODS: The sample was composed of 173 healthy children and adolescents, 96 (55.5%) F and 77 (44.5%) M, with ages between 10 and 16 years (mean: 13.04 +/- 1.68). The ACP activity was determined through a spectrophotometric method and its phenotypes through isoelectric focusing electrophoresis. BMI (Kg/m2) and the BP were obtained by standardized methods. Glycemia determined by the glucose oxidase method and insulinemia by RIA method. Insulin resistance based on the homeostasis model assessment (HOMA) was calculated as: [fasting insulin (microU/ml) x fasting glucose (mmol/l)]: 22.5. The %TFM and TF were determined by dual energy x-ray absorptiometry (DXA). The statistical methods used were ANOVA, the Pearson correlation and the Student's test. RESULTS: The distribution of the phenotypes were the following--absolute versus relative frequencies: BB-74 (48.4%), AB-52 (34%), AA-16 (10.5%), BC-7 (4.6%), AC-3 (2%) and CC-1 (0.7%). ACP activities (mean: 321.04 +/- 84.56) were significantly different among the phenotypes (p < 0.001). The smallest activity was observed in the AA individuals, the highest in CC, followed by BC (247.17 +/- 66.52 and 767.30 and 362.44 +/- 91.56 respectively). Glycemia was higher in the AA individuals (4.61 +/- 0.37) compared to CC + BC (4.40 +/- 0.31) (p = 0.08). A direct correlation was verified between HOMA and BP, both diastolic (p = 0.013, r = 0.250) and systolic (p = 0.015, r = 0.246), as well as of these with BMI (mean: 20.57 +/- 3.24) and insulinemia (p = 0.016, r = 0.215; p = 0.004, r = 0.280 and p = 0.007, r = 0.240; p = 0.008, r = 0.261 respectively for diastolic and systolic BP). There were no significant difference of BMI between sexes, nor of this as well as of % TFM and TF among the different genetic phenotypes of ACP. CONCLUSIONS: The smallest enzymatic activity of ACP seems to be associated with the AA individuals, where a trend for higher glycemia was verified. BMI, HOMA and insulinemia, due to their significant direct relationship with diastolic and systolic BP in this sample of children and adolescents may warrant more future attention in the evaluation of cardiovascular risk. There were no significant differences of HOMA, BMI, %TFM, TF nor of BP among the different ACP genetic phenotypes. PMID- 11941904 TI - [Basic principles of selection and use of diagnostic tests: practical applicability to individual patients]. AB - In clinical practice, in order to design and implement a specific therapeutic plan, as well as communicating an appropriate prognosis, the doctor needs to establish a precise diagnosis of the condition. Sometimes all one needs is a clinical impression. More often, however, the definition of an accurate diagnosis will mandate the interpretation of specific diagnostic tests as well. The rational use of diagnostic tests in cardiology--whether laboratorial or imaging- should be based on three factors: 1) validity of the study's results about the test; 2) diagnostic properties of the test and 3) applicability of the test in clinical setting. The rational use and correct interpretation of diagnostic tests are based on these three factors. In two previous articles, we presented the basic principles concerning the validity of the results from the study that defined the specific test as well as the diagnostic properties of the tests. In this final article we discuss the applicability of the test to the individual patient in clinical practice. PMID- 11941903 TI - Gender differences in the treatment of hypertension: a community based study in Porto. AB - OBJECTIVES: To describe the use of antihypertensive drugs in a random sample of adults living in Porto, Portugal, and to identify gender differences in the treatment of hypertension. DESIGN: Observational, cross-sectional. PARTICIPANTS AND METHODS: Nine hundred and fifty-nine participants over 39 years of age, living in Porto, were selected by random digit dialing. For each subject, socioeconomic characterization, family and personal medical history, and information on antihypertensive treatment were obtained through a questionnaire. Blood pressure was measured on a single occasion, and a fasting blood sample was collected. Gender differences in the treatment of arterial hypertension and number and type of drugs were evaluated through the calculation of female:male proportion ratios and 95% confidence intervals (95% CI). RESULTS: Hypertension treatment was more frequent in women than in men (proportion ratio 1.40, 95% CI 1.15-1.72), although no differences were observed among hypertensives aware of their condition (proportion ratio 1.07, 95% CI 0.93-1.22). The female:male proportion ratios of treatment with one drug, fixed combination therapy and free combination therapy were 1.13 (95% CI 0.94-1.36), 0.83 (95% CI 0.34-2.01) and 0.76 (95% CI 0.49-1.19), respectively. In subjects treated with one drug the use of ACE inhibitors/AT-II antagonists was more frequent in men (proportion ratio 0.68, 95% IC 0.46-1.01) and treatment with diuretics higher in women (proportion ratio 1.83, 95% CI 1.04-3.23). In participants treated with combination therapy, ACE inhibitors/AT-II antagonists and diuretics were more frequently used by women and calcium channel blockers and beta-blockers by men (female:male proportion ratios were 1.27, 95% CI 0.96-1.68, 1.24, 95% CI 0.94-1.64, 0.61, 95% CI 0.37 1.02 and 0.74, 95% CI 0.31-1.79, respectively). CONCLUSIONS: Arterial hypertension tended to be more frequently treated among women and different therapeutic options were found according to gender. Gender differences in the awareness of hypertension, sexual specificity of the activity of antihypertensive drugs, and comorbidity may play a role in gender inequalities in the treatment of hypertension in Portugal. PMID- 11941905 TI - [Ebstein's anomaly in an adult patient -- transesophageal echocardiography imaging]. PMID- 11941906 TI - Unlike my "brother," you are "heavy". PMID- 11941907 TI - Loss prevention case of the month. Questions of competence. PMID- 11941908 TI - Why don't the uninsured have health insurance? PMID- 11941909 TI - An interesting cause of lower gastrointestinal bleeding in the elderly. PMID- 11941910 TI - Fosinopril-induced hepatotoxicity in a complex medical patient. PMID- 11941911 TI - Expert testimony linking child sexual abuse with posttraumatic stress disorder. PMID- 11941912 TI - Insights on telemedicine: how big is the market? PMID- 11941913 TI - Healthcare IT and value with a HIPAA twist. PMID- 11941914 TI - HIPAA compliance: a technology perspective. PMID- 11941915 TI - Expanding beyond radiology. PMID- 11941916 TI - Planning and implementing HIPAA compliance activities. AB - This article reviews one healthcare network's HIPAA compliance management approach, detailing findings and trends from the assessment and presenting possible solutions derived from the remediation analysis activities, including fiscal and implementation timing considerations. PMID- 11941917 TI - HIPAA Readiness Collaborative in Hawaii. AB - The vision of Hawaii's HIPAA Readiness Collaborative (HRC) effort is to realize the positive potential of HIPAA through a collaborative process that engages the entire healthcare delivery system. Goals include reducing the cost of healthcare through streamlining, reducing the cost of HIPAA implementation for HRC participants, and improving the interoperability between facilities through use of standard technologies. PMID- 11941919 TI - Comparing HIPAA practices in two multi-hospital systems. AB - The comparison of HIPAA compliance efforts at two multi-hospital networks (Children's Health System and Carilion Health System) suggests a framework for classifying practices and for facilitating the integration of further case studies into a national library of HIPAA practices. PMID- 11941918 TI - A review of CHIN initiatives: what works and why. AB - A Community Health Information Network (CHIN) can generate revenue through multiple avenues and maximize efficiency for participants, including multiple payers, physicians, hospitals, labs, pharmacies, and ancillary providers. This article reviews representative CHIN initiatives over the past decade and identifies key success factors, such as provider adoption and participation in planning, multiple-payer participation, providing key transactions, and incremental implementation. PMID- 11941920 TI - Benchmarking HIPAA compliance. AB - One of the nation's largest academic medical centers is benchmarking its operations using internally developed software to improve privacy/confidentiality of protected health information (PHI) and to enhance data security to comply with HIPAA regulations. It is also coordinating the development of a web-based interactive product that can help hospitals, physician practices, and managed care organizations measure their compliance with HIPAA regulations. PMID- 11941921 TI - Mobilizing the right resources to achieve HIPAA compliance. PMID- 11941922 TI - Real-time management of IT. AB - The ability to successfully manage information technology-driven transformations of work processes into value--which has always been a critical requirement yet often a huge disappointment--is now becoming a main ingredient of success in all industries, including healthcare. This article prescribes management dimensions that healthcare leaders should focus on to improve their ability to manage and derive value from the re-invented work flow, especially the highly visible opportunities offered by e-commerce and the consumer movement. PMID- 11941923 TI - Managing non-technical factors in healthcare IT projects. AB - The implementation of IT within any healthcare organization is a demanding and intense exercise in the management of both technical (systemic) and non-technical (social) influences and factors, and the project process is often fraught with unforeseen obstacles or impediments to success. Project success depends as much on a specific understanding of the non-technical project impediments such as conflicting personalities and corporate culture as it does on the performance of the applied technology or the viability of the implementation process itself. PMID- 11941924 TI - IT investment planning: the best hospitals. AB - A study of hospitals' best practices regarding capital investments in information systems indicates that (1) the process may need more input from IT professionals to cover the technical aspects of planning; (2) the most successful projects are highly aligned with the organization's overall business strategy; and (3) disappointments could be avoided by conducting a better analysis of the organization and its technology, evaluation of past IS performance, and assessment of IS staff capabilities. PMID- 11941925 TI - Gaining MD buy-in: physician order entry. AB - Computerized physician order entry (CPOE) eliminates illegible handwriting, reduces medical errors, and improves patient care. The administration, medical staff, nursing, and health information systems departments of a community teaching hospital cooperated to achieve organization-wide use of its CPOE system. PMID- 11941926 TI - A physician's perspective: deploying the EMR. AB - This case study presents a collection of thoughts regarding the process of selecting and deploying an electronic medical record (EMR) in a physician's practice. Practical advice and observations are detailed about what was learned and how the practice has benefited. PMID- 11941927 TI - Managing Web access through delegated administration. PMID- 11941928 TI - Computer games may be good for your health. AB - Computer-based games and interactive simulators have matured to the point where they can have a significant impact on healthcare behavior decisions. Initial results from using one such game (Heart Sense) indicate that it can improve recognition of heart attack symptoms and shift behavioral issues so as to reduce pre-hospitalization delay in seeking treatment--thereby reducing myocardial infarction mortality and morbidity. PMID- 11941929 TI - Malignancy in choledochal cysts. AB - BACKGROUND/AIMS: Malignancy in choledochal cysts is a rare condition. This study presents our experience with this condition, with emphasis on the clinical presentation, management and outcome. METHODOLOGY: Subjects included 80 adults with choledochal cysts treated from January 1979 to December 1995. Of these patients, eight were found to have malignancy in the cyst and formed the basis of this study. RESULTS: Four patients had synchronous and four had metachronous carcinoma lesions arising in the choledochal cyst. The clinical presentations were: biliary tract infection in 5 patients, gastric outlet obstruction in two and right upper quadrant pain and body weight loss in one. Operations for bile duct malignancy included total excision in two patients, choledochotomy with T tube drainage in two patients, gastrojejunostomy in two patients, percutaneous transhepatic biliary drainage and gastrojejunostomy in one patient and metastatic lymph node biopsy only in one. One patient died due to septic shock within 30 days of operation (operative mortality). Postoperative survival time ranged from 4 to 9 months with a mean of 6.2 months. One of these patients was still alive at the time of writing (7 months). CONCLUSIONS: The frequency of malignancy in the choledochal cysts was 10% in our series. Malignancy in the choledochal cyst should be highly suspected in patients with cholangitis symptoms, body weight loss and anemia. Prognosis in this disease entity is poor. PMID- 11941930 TI - Studies on biliary tract carcinoma in the case with pancreaticobiliary maljunction. AB - BACKGROUND/AIMS: The purpose of this study was to clarify the clinicopathological features of pancreaticobiliary maljunction and to determine the appropriate surgical approach for biliary tract with pancreaticobiliary maljunction. METHODOLOGY: The data of 77 patients with pancreaticobiliary maljunction including 13, who had been treated for biliary tract cancer, were reviewed retrospectively. We assessed the clinical features, biological characteristics of the cancer, methods of surgical treatment, postoperative outcome and cell proliferating activity of the biliary epithelium, evaluated by the PCNALI (proliferating cell nuclear antigen-labeling index). RESULTS: The incidence of cancer development in the case with pancreaticobiliary maljunction was 13.4% in the bile duct dilatation group (n = 67) and 40.0% in the non-dilatation group (n = 10). Dissection of lymphadenectomy was performed in 10 (76.9%) of 13 patients, and curative resection was feasible in 9 of the 10 patients. Two (20.0%) of the 10 patients had lymph node involvement noted at surgery and died of recurrence. In the other eight patients without lymph node involvement at surgery, six patients underwent curative resection and are alive at 7 months to 11 years and 6 months after surgery. PCNALI of the biliary epithelium of the patients with pancreaticobiliary maljunction was significantly higher than that of the control group. CONCLUSIONS: For patients with pancreaticobiliary maljunction, it should be stressed that the extrahepatic bile duct be prophylactically removed, even when there are no neoplasmatic changes because of high prevalence of cancer development, presumably predicted by the increase of cell proliferative activity in the biliary epithelium. For patients with biliary cancer, early detection at the stage with no lymph node involvement is essential to secure for long-term survival. PMID- 11941931 TI - Double cancer of gallbladder and bile duct associated with anomalous junction of the pancreaticobiliary ductal system. AB - We report a case of double cancer of the gallbladder and the common bile duct associated with anomalous junction of the pancreaticobiliary ductal system, and review the literature of similar case reports. A 66-year-old woman was admitted to an associated hospital complaining of upper abdominal pain, and was diagnosed as having pancreatitis. Abdominal imaging revealed an irregularly protruding mass at the body of the gallbladder and an intraluminal protrusion at the lower third of the common bile duct. Endoscopic retrograde cholangiopancreatography also revealed anomalous junction of the pancreaticobiliary ductal system with congenital biliary dilatation of 14 mm in the largest diameter. She underwent surgical resection of the gallbladder, the extrahepatic bile duct and the gallbladder bed of the liver with a dissection of the regional lymph nodes for double cancer of the gallbladder and the bile duct associated with anomalous junction of the pancreaticobiliary ductal system. She is still alive 33 months after surgery without any signs of recurrence. There were 12 patients (including our case) reported in the literature who had double cancer of the gallbladder and the extrahepatic bile duct associated with anomalous junction of the pancreaticobiliary ductal system. Only 33% of these 12 patients had jaundice. Tumors of the 12 patients were commonly early-stage cancer both in the gallbladder (36%) and in the extrahepatic bile duct (73%). Therefore, we concluded that precise preoperative imaging of the total biliary tract should be required in order to detect early-stage cancer in patients with anomalous junction of the pancreaticobiliary ductal system before planning surgical procedures, and consideration should be given to the possibility of multiple occurrences of biliary tract cancers. PMID- 11941933 TI - Immunohistochemical localization of vascular endothelial growth factor in colonic mucosa of patients with inflammatory bowel disease. AB - BACKGROUND/AIMS: Significantly enhanced serum levels of VEGF (vascular endothelial growth factor) were found in patients with inflammatory bowel disease. Peripheral blood mononuclear cells have been identified as one of the origins of the circulating VEGF. The present investigation examines the localization of VEGF at the site of inflammation in colonic mucosa of patients with Crohn's disease and ulcerative colitis. METHODOLOGY: Immunohistochemical localization of VEGF and immunostaining for leukocytes were performed in colonic mucosal biopsies of 41 patients with Crohn's disease, 26 patients with ulcerative colitis and normal mucosal specimens of 5 patients with irritable bowel syndrome. Measurement of immunohistochemical staining for VEGF and for leukocytes within the epithelium and the lamina propria was performed separately by area morphometry using a computerized cell analysis system. RESULTS: In both patients with Crohn's disease and ulcerative colitis immunohistochemical staining for VEGF within the lamina propria of inflamed colonic mucosa was significantly higher compared with noninflamed mucosa (Crohn's disease: 4.26% vs. 0.07%, P < 0.001; ulcerative colitis: 3.68% vs. 0.32%, P = 0.001). There was a significant correlation between immunostaining for leukocytes and VEGF within the lamina propria in both patients with Crohn's disease (r = 0.73, P < 0.05)) and ulcerative colitis (r = 0.67, P < 0.05). In Crohn's disease immunostaining for VEGF within the epithelium was significantly higher in inflamed mucosa compared with noninflamed mucosa (9.85% vs. 0.63%, P < 0.001). In contrast, strong immunostaining for VEGF has been observed in the epithelium of noninflamed mucosa (7.60%, P < 0.003), as well as in inflamed mucosa of patients with active ulcerative colitis (9.68%, P < 0.002) compared with noninflamed mucosa of patients with inactive ulcerative colitis (1.39%). CONCLUSIONS: The present data indicate, that the increased VEGF expression within the epithelium and the interstitial accumulation of VEGF-producing leukocytes in inflamed mucosa may play an important role in the inflammatory mechanisms of Crohn's disease and ulcerative colitis. PMID- 11941932 TI - Percutaneous management of bile duct injuries after cholecystectomy. AB - Intraoperative biliary tract injuries are relatively uncommon, but are a cause of significant morbidity and mortality. We have repaired open biliary tract injuries found postoperatively in three patients without reoperation by percutaneous intervention. In two cases, biliary stents were placed percutaneously across the injured portion of the bile duct. For this procedure, a unique coaxial guidewire technique was used. The bile leaks resolved soon after insertion of the biliary stents. In one patient, an expandable metallic biliary endoprosthesis was inserted, and the temporary stent was removed. This patient had no signs or symptoms of cholangitis or biliary obstruction at the 6-year follow-up. The other patient died of heart failure soon after reconstruction. In the last patient, a biliary drain was inserted through the injured duct via the biliocutaneous fistula. In this case, biliary drainage alone resulted in resolution of the bile leak, because the injury was partial without a stricture. This patient was well at the 3-year follow-up. Percutaneous management of bile duct injuries is an alternative in selected patients. PMID- 11941934 TI - The European approach to hepatocellular carcinoma. AB - Patients with cirrhosis of viral, metabolic or autoimmune origin are at high risk of developing hepatocellular carcinoma. Prospective surveillance based on semi annual ultrasound examination of the abdomen has allowed for detection of small tumors in many patients, but it is not clear whether liver-related mortality was decreased in parallel. Prognostication in patients with hepatocellular carcinoma requires integrated assessment of tumor size and number, liver function and performance status. The therapeutic approach is to a large extent non-evidence based and the best treatment choice depends on individual patients characteristics, taking into account the local technological and therapeutic resources and skills. Since surgical resection, liver transplantation and percutaneous ablation have achieved a high rate of complete response in properly selected patients, these procedures are considered curative treatments. Being curative treatments applicable only to patients with a small tumor, hepatocellular carcinoma surveillance aimed at early detection of the tumor is the most practical approach for improving treatment outcome. PMID- 11941935 TI - Colorectal polyp detection with virtual colonoscopy (computed tomographic colonography); the reliability of the method. AB - BACKGROUND/AIMS: The aim of this study was to compare the reliability of virtual colonoscopy with conventional colonoscopy in detection of colorectal polyps. METHODOLOGY: Between November 1997 and February 2000, 23 patients (14 males, 9 females) aged 43-86 years (mean: 63) with colorectal polyps who underwent colonoscopy in our endoscopy unit, were referred for virtual colonoscopy. The colon was distended with an enema tube for rectal insufflation and subsequently was scanned by helical CT (Toshiba X-vision) using the following parameters: KV 120, mAs 200, table feed 5 mm/sec, slice thickness 5 mm and reconstruction intervals 2.5 mm. Using SPARC Station 20, virtual colonoscopy was generated. RESULTS: A total of 30 polyps (19 with diameter 8-15 mm and 11 larger than 15 mm) were detected with conventional colonoscopy. Virtual colonoscopy visualized 27 polyps (17 with diameter 8-15 mm and 10 with diameter > 15 mm). CONCLUSIONS: Virtual colonoscopy is a non-invasive, well-tolerated, safe technique and can be useful for detection of colorectal polyps. Weaknesses of the method are the inability to detect small lesions and to perform biopsies. PMID- 11941936 TI - Prevalence of Helicobacter pylori in the residual stomach after gastrectomy for gastric cancer. AB - BACKGROUND/AIMS: In recent years, the role of Helicobacter pylori in gastritis of the residual stomach has attracted much attention. We investigated the prevalence of Helicobacter pylori in the residual stomach after distal gastrectomy for gastric cancer, as well as the correlations between Helicobacter pylori positivity and clinical characteristics or the severity of gastritis in the residual stomach. METHODOLOGY: The subjects were 66 patients with gastric cancer who underwent distal gastrectomy with Billroth I reconstruction at our department. Helicobacter pylori was detected by the 13C-urea breath test, and patients were considered to be Helicobacter pylori-positive if the delta 13C value was > 2.5@1000. RESULTS: The overall Helicobacter pylori positivity rate of the gastrectomy patients was a high 80.3%, with the rate being especially high in patients under 60 years of age and in those tested less than 5 years after surgery. There was a close relationship between Helicobacter pylori positivity and the severity of gastritis. CONCLUSIONS: Helicobacter pylori infection appears to cause the development of gastritis. Helicobacter pylori eradication needs to be taken into consideration in the management of Helicobacter pylori-positive patients after gastrectomy. PMID- 11941937 TI - Connective tissue growth factor is directly related to liver fibrosis. AB - BACKGROUND/AIMS: Connective tissue growth factor is a mitogenic, chemotactic, and cell matrix-inducing factor for fibroblasts, and is exclusively and directly induced by transforming growth factor-beta 1. In skin fibrosis, connective tissue growth factor is thought to be a downstream effector of transforming growth factor-beta 1 that is directly involved in the proliferation of connective tissue cells and the accumulation of matrix. Our aim is to confirm the involvement of connective tissue growth factor in liver fibrosis. METHODOLOGY: Specimens obtained from autopsy of 12 patients with normal liver (n = 3), hepatocellular carcinoma (n = 3), liver cirrhosis secondary to alcoholic abuse (n = 3), or viral hepatitis (n = 3), were used for digoxigenin labeled in situ hybridization. RESULTS: Abundant connective tissue growth factor messages were detected in the fibrotic area between cirrhotic nodules. Hepatocytes did not show any signals even when they became carcinomas. Normal livers showed few or no signals. CONCLUSIONS: These results confirmed the direct relationship between connective tissue growth factor gene expression and liver fibrotic change. PMID- 11941938 TI - Changes of hepatic sinusoids in experimental septicemia of rats are correlated with the production of nitrogen monoxide. A morphometrical and immunohistochemical study. AB - BACKGROUND/AIMS: Two types of experimental septic models were prepared to morphologically and histometrically analyze the changes in the hepatic sinusoid. Furthermore, the local expression of nitrogen monoxide synthetase was analyzed by immunohistochemical and immune electron microscopical studies for the purpose of investigating the involvement of nitrogen monoxide in these changes. METHODOLOGY: Using Sprague-Dawley strain male rats, two types of experimental septic models were prepared. These were a bacterial peritonitis model induced by a cecal ligation and puncture method, and a peritonitis model after intraperitoneal administration of endotoxin at 10 mg/kg. Serum alanine aminotransferase and endotoxin were measured in due order. The liver tissues were sampled and examined by usual light microscopical and electron microscopical analyses. The sites of the portal and central vein regions were randomly selected, and the sinusoidal cavity/liver volume ratio was estimated by the point counting method under high magnification. In addition, the expressions of nitrogen monoxide synthetase were investigated in the collected samples of the liver, lung and aortic wall. RESULTS: According to the histometrical analysis of the sinusoid, the sinusoidal cavity volume ratio at 6 hours after the onset increased from 0.0690 +/- 0.0147 to 0.089 +/- 0.004 in the central vein region. The ratio rapidly increased from 0.058 +/- 0.009 to 0.093 +/- 0.008 in the portal region from 6 to 12 hours after the onset. The values of serum alanine aminotransferase and endotoxin were significantly increased from 6 to 12 hours after the onset of peritonitis. The inducible nitrogen monoxide synthetase was observed in the alveolar macrophages, monocytes, and hepatic Kupffer cells from 6 to 12 hours after the onset of peritonitis. CONCLUSIONS: These results demonstrated that the sinusoidal cavity became dilated in the sepsis model and at the same time nitrogen monoxide synthetase was expressed in the sinusoidal wall-composing cells. There is a possibility that the action of nitrogen monoxide is involved in the changes in the volume of the sinusoidal cavity, suggesting the involvement of nitrogen monoxide derived from sinusoidal wall-composing cells in the hepatic dysfunction caused by sepsis in addition to nitrogen monoxide generation from Kupffer cells. PMID- 11941939 TI - Left hepatectomy for the choledochal cyst (type IV-A) with intrahepatic stenosis: report of a case. AB - The case of a 16-year-old male with expansion of the gallbladder and dilatation of the common bile duct is reported. Ultrasonography and computed tomography imaging showed expansion of the gallbladder and eminent cystic dilatation in the common bile duct and the left intrahepatic bile duct. Endoscopic retrograde cholangiopancreatography indicated expansion-like beads of the bilateral hepatic ductus and the left intrahepatic bile duct, including anomalies of the pancreaticobiliary ductal junction. Because relative stenosis of the membranous diaphragm was revealed in the porta hepatis, we diagnosed this case as a type IV A choledochal cyst, using Todani's classification. Intraoperative cholangiography and cholangiofiberscopy showed a pinhole stricture and re-expansion of the tip of the left intrahepatic bile duct. As the narrow segment could not be expanded though we put proper pressure there, left hepatectomy was performed as a preventive measure in addition to extended biliary tract excision and cholangiojejunostomy. Hepatectomy seems to be an appropriate choice in a case of intrahepatic stenosis to help increase the patient's postoperative quality of life. PMID- 11941940 TI - Spleen-preserving laparoscopic distal pancreatectomy for cystic adenoma. AB - For borderline malignant diseases of the pancreas such as cystic adenoma, partial pancreatectomy or pancreatoduodenectomy including pylorus-preserving pancreaticoduodenectomy have been performed depending on tumor location under large median laparotomy. To investigate the feasibility of a technique with minimal skin incision, while retaining safety equivalent to conventional resection of the pancreatic tail, by making use of the advantages of laparoscopic procedure, we performed a minimally invasive laparoscopic resection of the pancreatic tail with preservation of the spleen. A 69-year-old woman underwent surgery for a diagnostic therapy for a cystic lesion of the pancreatic tail. The procedure was performed as follows: All procedures were performed completely laparoscopically under CO2 insufflation. After dissection of the omentum, laparoscopic ultrasound was performed to identify the location of the tumor and splenic vessels. The splenic hilus was dissected with preservation of the splenocolic ligament to maintain the lower blood supply to the spleen. The left gastroepiploic artery and the short gastric arteries and veins could be preserved. After division of the splenic hilus, the splenic artery and vein were identified from behind the pancreas by being held up and dissected individually by intracorporeal ligation by 3-0 Nylon. Then, pancreatic transection was performed 1 cm proximal to the tumor with the Endo-GIAII. The duration of operation was 4.5 hours. Intraoperative blood loss was under 50 mL. Histological examination revealed mucinous cytadenoma. She could walk the day after surgery and was discharged from the hospital uneventfully. CT prior to discharge from the hospital revealed sufficient blood flow in the spleen. Thus, it may be feasible to select laparoscopic spleen-preserving distal pancreatectomy as a first choice for diagnostic therapy for cystic lesions of the pancreatic tail. PMID- 11941941 TI - Advanced gastric cancer associated with DIC successfully treated with 5-FU and cisplatin: a case report. AB - We report a case of advanced gastric cancer complicated by disseminated intravascular coagulation successfully treated with chemotherapy consisting of 5 fluorouracil and cisplatin. The patient was a 53-year-old woman who complained of loss of appetite, weight loss, and low back pain. Based on the laboratory data, a diagnosis of disseminated intravascular coagulation was made. Gastroscopy revealed gastric carcinoma (Borrmann type 3) that was continuously bleeding, and chest computed tomography showed a solitary lung metastasis and bilateral pleural effusion. Bone scintigraphy revealed multiple bone metastases. Accordingly, we made a diagnosis of stage IV gastric cancer complicated by disseminated intravascular coagulation. We selected the 5-fluorouracil and cisplatin combination chemotherapy for treatment and obtained the patient's consent. After two cycles of the 5-fluorouracil and cisplatin therapy, the bleeding symptoms improved and the disseminated intravascular coagulation process was successfully controlled. We concluded that disseminated intravascular coagulation caused by gastric cancer may be improved when the primary cancer and its metastases are brought under control by treatment with FP combination chemotherapy. PMID- 11941942 TI - Colorectal cancer risk in first-degree relatives of patients with colorectal adenomatous polyp. AB - BACKGROUND/AIMS: To evaluate any risk of colorectal cancer in first-degree relatives of patients with colorectal adenomatous polyp. METHODOLOGY: In a screening program-based cross-sectional study, 44821 subjects received an immunochemical fecal occult blood test using a 2-consecutive-day method. They were divided into two groups, according to the results of a self-completed questionnaire on family history of colorectal adenomatous polyps, and the positivity rate of an immunochemical fecal occult blood test as well as the positive predictive value for colorectal cancer were determined in these two groups. RESULTS: The fecal occult blood test was positive in 8.5% of subjects with family history and in 4.8% of subjects without family history, and the positive predictive value for colorectal cancer was 6.8% and 2.4% in subjects with and without family history of colorectal adenomatous polyps, respectively, indicating a significant difference in the positivity rate of the fecal occult blood test (P < 0.01) as well as the positive predictive value for colorectal cancer (P < 0.05) between these two groups. CONCLUSIONS: These results show that first-degree relatives of patients with colorectal adenomatous polyp have an increased risk for colorectal cancer, and that the subjects with family history of colorectal adenomatous polyps as well as cancers should be considered as a priority group for prevention of colorectal cancer. PMID- 11941943 TI - CA19-9 as the most significant prognostic indicator of metastatic colorectal cancer. AB - BACKGROUND/AIMS: Colorectal cancer is the third leading cause of cancer-related mortality in Taiwan. We became interested in searching for the factors predictive of survival. Serum CA19-9 (carbohydrate antigen 19-9) level has been reported as a factor predictive of survival in patients with colorectal cancer. A few articles have reported that patients with metastatic colorectal cancer who have normal (< or = 37 U/mL) serum CA19-9 levels survived significantly longer than those with higher serum CA19-9 levels. However, these reports are contradictory and lack definite conclusions. This study was carried out in an effort to evaluate the prognostic significance of serum CA19-9 levels in patients with metastatic colorectal cancer in Taiwan. METHODOLOGY: Between 1991 and 1994, a total of 128 patients with histologically confirmed metastatic colorectal cancers were evaluated retrospectively at Veterans General Hospital-Taipei. All patients had measurable metastatic lesions and life expectancies of more than 3 months. 5 Fluorouracil-based chemotherapy, either in a weekly bolus regimen or a monthly 5 day bolus schedule, were administered to all of them. Data on age, sex, performance status, location of primary tumor, extent of metastases, site of metastases, histological differentiation, serum CEA (carcinoembryonic antigen) and CA19-9 levels were analyzed before chemotherapy to determine their association with survival. Blood samples for CEA and CA19-9 measurement were analyzed using the radioimmunoassay method. Multivariate analysis by the Cox's proportional hazards regression model was performed to determine independent prognostic factors among all of the possible variables. RESULTS: By univariate analysis, serum CA19-9 levels (P < 0.001) and performance status of the patients (P = 0.022) were identified as prognostic factors, while age, sex, location of primary tumor, site of metastasis, histological differentiation, and pre treatment serum CEA levels were not considered significant. By multivariate analysis, serum CA19-9 levels (P < 0.001) and performance status of the patients (P = 0.014) were still found as independent prognostic factors of these patients. CONCLUSIONS: The data from our study indicate that serum CA19-9 level is the most significant prognostic indicator of patients with metastatic colorectal cancer. It is recommended that stratification for further clinical trials for patients with metastatic colorectal cancer should be carried out according to serum CA19-9 levels. PMID- 11941944 TI - Doppler US of superior mesenteric artery in the assessment of ulcerative colitis. A prospective study. AB - BACKGROUND/AIMS: The purpose of this study was to correlate blood flow velocity in the superior mesenteric artery and activity of ulcerative colitis. METHODOLOGY: Doppler spectral analysis of superior mesenteric artery blood flow velocities were obtained in a blind study from 28 patients after fasting (A1: 13 patients with pancolitis, A2: 5 patients with subtotal colitis, B: 10 patients with left-sided colitis) and 50 healthy volunteers (control group). Disease activity was determined with clinical and endoscopic findings. RESULTS: A significant increase in superior mesenteric artery blood flow measurements was observed in the active pancolitis group A1 [Vsyst = 3.64 +/- 0.18 m/sec and Vdiast = 0.94 +/- 0.09 m/sec as compared with healthy volunteers (Vsyst = 1.14 +/ 0.07 m/sec, Vdiast = 0.38 +/- 0.04 m/sec) P < 0.01. A minor increase in superior mesenteric artery blood flow velocity was observed in patients with subtotal colitis, group A2 (Vsyst = 2.06 +/- 0.14 m/sec, Vdiast = 0.45 +/- 0.05 m/sec) as compared with healthy volunteers P < 0.01. In group B with left sided colitis superior mesenteric artery velocity changes were not statistically significant (P > 0.05). CONCLUSIONS: Doppler US velocity measurement of superior mesenteric artery may be used as an adjunct in the assessment of ulcerative colitis extension and activity. PMID- 11941945 TI - Methods and related drawbacks in the estimation of surgical risks in cirrhotic patients undergoing hepatectomy. AB - There has been a dramatic improvement in recent results of hepatectomy for hepatocellular carcinoma in cirrhotic patients. Hospital mortality rates of less than 5% are frequently reported. The improvement is largely a result of better techniques and performance of surgeons in hepatectomy, and reduction in blood loss and transfusion requirement. Better selection of patients is perhaps a more significant contributory factor. Careful identification of risk factors related to the medical condition of the patient, functional reserve of the liver and volume of the remnant liver is essential for the prevention of postoperative liver failure. Indocyanine green clearance test is the most accurate test for assessment of liver function reserve. An indocyanine green retention rate of 14% at 15 minutes is the safety limit for major hepatectomy for cirrhotic patients. A maximum of 60% of the nontumorous liver can be resected safely. Computed tomography is therefore an important assessment parameter. The liver function reserve also reveals the suitability for hepatectomy. Liver enzymes, alanine aminotransferase or aspartate aminotransferase can reflect the hepatic activity, which could be responsible for the impaired liver function. Steatosis is another factor that influences hepatic function reserve. Age is also an important risk factor in hepatectomy because elderly patients may harbor occult heart disease, reduced respiratory and liver function reserves. After recognizing the risk factors, surgeons should eliminate operative morbidity and mortality by making appropriate decisions based on the assessments. In conclusion, preoperative risk assessment involves evaluation of hepatic function reserve, remnant liver volume, liver status, age and the medical condition of the patient. A 0% hospital mortality rate is considered the objective. PMID- 11941946 TI - Dual versus triple therapy in eradication of Helicobacter pylori. AB - BACKGROUND/AIMS: Duodenal ulcers should be treated by eradication of Helicobacter pylori. This study compared the efficacy of a proton pump inhibitor together with one or two antibiotics in eradication therapy. METHODOLOGY: 177 patients who were H. pylori positive were randomized to receive 14 days of either: lansoprazole 30 mg bd and amoxicillin 1 g bd (LA), omeprazole 20 mg bd and amoxicillin 1 g bd (OA) or lansoprazole 30 mg bd, amoxicillin 1 g bd and clarithromycin 500 mg bd (LAC). The efficacy was assessed at four weeks and at six months after the end of treatment. Biopsies were taken for culture and bacterial sensitivity testing at inclusion and at four weeks after the end of treatment. RESULTS: 149 patients were evaluated for efficacy. The eradication rate was significantly higher in LAC (96%) compared to LA (51%) and OA (64%) treatments (P < 0.001). At baseline 17%, 21% and 19% of the patients in the LA, OA and LAC groups, respectively, were resistant to metronidazole and only one patient was resistant to clarithromycin. Post-treatment, four patients had acquired metronidazole resistance. CONCLUSIONS: LAC is more effective than LA and OA for eradication of H. pylori in duodenal ulcer disease. PMID- 11941947 TI - How many lymph nodes are needed for an accurate pN classification in esophageal cancer? Evidence for a new threshold value. AB - BACKGROUND/AIMS: The UICC recommends a number of at least six lymph nodes to be examined in the surgical therapy of esophageal cancer for a reliable pN classification. The aim of this study was to evaluate this threshold by means of the data from our patients. METHODOLOGY: Following curative resection (R0) of esophageal cancer the numbers of examined tumor-free and tumor-involved lymph nodes were compared. Different statistical models of logistic regression were fitted to the data and checked for plausibility (Hosmer Lemeshow test). The sensitivity of a correct pN classification was then calculated and correlated to the total number of examined lymph nodes. RESULTS: A maximum increase of the sensitivity in classifying pN occurred from 0 to 6 examined lymph nodes. Nevertheless an additional improvement of sensitivity was continuously shown up to 100 examined nodes. An over 90% sensitivity of a correct lymph node classification was reached when more than twelve nodes were examined. Thus the results demonstrate in the case of esophageal cancer, that the suggestion by the UICC to examine at least 6 nodes for defining pN appears too low and may not represent the clinical situation. A ninety percent confidence level of a correct lymph node classification can be expected above 12 examined nodes similarly to the current recommended threshold in colorectal carcinoma. CONCLUSIONS: We suggest a new threshold for the number of examined lymph nodes of at least 12 instead of 6 nodes for accurately defining the pN category in esophageal cancer. PMID- 11941948 TI - Omentoplasty versus no omentoplasty for cervical esophagogastrostomy following radical esophagectomy. AB - BACKGROUND/AIMS: Omentoplasty--wrapping the omentum around the alimentary tract anastomosis is thought to lower the rate of anastomotic leakage. We evaluated the role of omentoplasty to reinforce cervical esophagogastrostomy after radical esophagectomy. METHODOLOGY: We compared anastomotic leakage, stricture formation, and related deaths in 63 patients who underwent radical esophagectomy and cervical esophagogastrostomy, with (n = 48) or without (n = 15) omentoplasty, between 1995 and 1999. RESULTS: An esophageal anastomotic leakage was diagnosed in 1 of the 48 patients (2.1%) with omentoplasty versus 3 of the 15 patients (20.0%) without omentoplasty (P < 0.01). Anastomotic stricture occurred in 2 (4.2%) of the omentoplasty group and 1 (6.7%) of the no omentoplasty group (P < 0.01). Death within 1 month was zero in the omentoplasty group and one (6.7%) in the no-omentoplasty group, despite no differences in lethal anastomotic leakage. CONCLUSIONS: Omentoplasty of cervical esophagogastrostomy reduced anastomotic leakage. Although promising, these observations require confirmation with a randomized prospective study. PMID- 11941949 TI - Clinical impact of patients with ruptured esophageal varices with death outcome. AB - BACKGROUND/AIMS: High hemostasis rates for ruptured esophageal varices have been acquired, but many patients die of liver failure afterwards. The aim of the current study is to find clinical information about the patients with ruptured esophageal varices who died during an early period after acquiring a hemostasis. METHODOLOGY: Forty-five patients with ruptured esophageal varices, including 17 who died during two months after acquiring a hemostasis by an initial endoscopic therapy, were investigated in the current study. The relationship between the clinical data and the prognosis of the patients were examined. RESULTS: The proportion of cases with Child-Pugh C in patients who died after therapy significantly higher than that in survivors (P < 0.01). The level of hemoglobin in patients who died was significantly lower than that in the survivors (P < 0.01). Moreover the proportion of an elevation of serum C-reactive protein in patients who died was significantly higher than that of the survivors (P < 0.05). CONCLUSIONS: Our investigation suggested that the elevation of serum C-reactive protein level and a devastated liver function influenced by hemorrhage may be indicators of unfavorable outcomes of the patients with ruptured esophageal varices. PMID- 11941950 TI - Results of Savary-Gilliard dilatation in the management of cervical web of esophagus. AB - BACKGROUND/AIMS: Cervical web is predominantly a disease of middle-aged females. Sixteen patients with cervical web of esophagus were studied. Dysphagia is mostly for solids, intermittent, non-progressive and was present for many months before they presented. METHODOLOGY: Treatment consisted of forceful dilatation with Savary-Gilliard bougies of 15 or 17 mm diameter after initial rupture of the web by endoscope or 11 mm dilator. RESULTS: Single session of dilatation was adequate in all patients (100%) and there was no recurrence of web during follow-up of 22.8 +/- 18.9 months. CONCLUSIONS: Endoscopic dilatation is a simple outpatient procedure having least morbidity and is suggested as the treatment modality of choice in the management of cervical web of the esophagus. PMID- 11941951 TI - Ruptured esophageal varices caused by metastatic liver tumors. AB - We herein report a case of a 50-year-old male with hematemesis due to the rupture from esophageal varices coexisting with multiple liver tumors metastasizing from sigmoid colon cancer. The causal etiology of esophageal varices rupture in the current case was considered to be the intrahepatic portal hypertension brought by the stagnation of the blood flow of the intrahepatic portal vein suppressed by the metastatic liver tumors. PMID- 11941952 TI - Efficacy of trimebutine therapy in patients with gastroesophageal reflux disease and irritable bowel syndrome. AB - BACKGROUND/AIMS: GERD (gastroesophageal reflux disease) occurs in 25-51% of IBS (irritable bowel syndrome) patients. Trimebutine has been effective in some IBS patients by modulating colonic motility. Furthermore, it increases gastric emptying rates, and controls esophageal motility. The aim of this study was to investigate the efficacy of trimebutine therapy in GERD patients with IBS. METHODOLOGY: Sixty-nine patients with GERD and IBS underwent upper gastrointestinal endoscopic, histologic and clinical evaluation prior to and 3 months post-treatment. H. pylori presence was determined by histology and CLOtest. Forty patients (Group A) were treated with omeprazole plus trimebutine for 3 months: in 32 H. pylori-positive patients (subgroup A1), a standard triple eradication regimen was introduced. Twenty-nine patients (Group B) were treated with omeprazole for 3 months: in 24 H. pylori-positive patients (subgroup B1), the same eradication therapy was employed. RESULTS: Specialized intestinal metaplasia of the gastroesophageal junction was observed in 20% and in 17.2% of the patients in Groups A and B, respectively. Eradication rates were similar in subgroups A1 (84%) and B1 (83%). In Group A there was a significant improvement in GERD (P = 0.003) and IBS symptoms (P < 0.0001) as well as esophagitis (P = 0.029), when compared with Group B. CONCLUSIONS: Trimebutine appears to be effective in patients with GERD and IBS. PMID- 11941953 TI - Imprint cytology in the endoscopic diagnosis of gastrointestinal malignancies. AB - BACKGROUND/AIMS: To compare imprint cytology with histopathology regarding diagnostic accuracies and quickness in the diagnosis of gastrointestinal malignancies. METHODOLOGY: Multiple endoscopic biopsies were taken from 146 patients having various lesions without a prior histopathological diagnosis. Imprint smears were prepared, using all biopsied tissues, on 3-5 slides. Tissues were fixed in 10% formalin. Time from the materials were received at the pathology laboratory to when the results were available was recorded. Cytopathologists and histopathologists, blinded to each other's diagnosis, examined the imprint smears and tissues. RESULTS: Average time to get imprint cytology results was shorter than that of histopathology (55 minutes vs. 8 days). Fifty-eight patients had a final diagnosis of malignancy. Histopathology and imprint were positive in 56 and 54, respectively. Eighty-eight patients had benign histopathology, all of these had negative imprint results. False-negative and false-positive rates for imprint were 6.8%, 0% and for histopathology were 3.4%, 0%, respectively. CONCLUSIONS: Imprint cytology is an easy, reliable method that can be used as an adjunctive measure with histopathology. It gives the earliest information about the nature of the lesion with a minimum misdiagnosis risk. Imprint cytology lets the surgeon plan a therapeutic strategy approximately 1 week earlier. PMID- 11941954 TI - An assessment of the effect of Ginkgo Biloba EGb 761 on ischemia reperfusion injury of intestine. AB - BACKGROUND/AIMS: The aim of this study was to determine the effect of Ginkgo Biloba (EGb 761) on reperfusion injury of the small bowel. METHODOLOGY: Forty eight male 200-250 g Spraque-Dawley rats in six groups were used to determine the biochemical and histopathological changes after a 30-min ischemia and 30-min reperfusion. Pre-treatment with 50 mg/kg EGb 761 (Tebofortan, Karlsruhe-Germany) or 10-mL/kg saline was administered intravenously in the treatment and control groups. The superior mesenteric artery was occluded distal to the right colic artery and collateral arcades were ligated to provide complete ischemia. Ischemia was determined by the existence of pulseless or pale color of the small intestine. The return of the pulses and the reestablishment of the pink color were assumed to be the reperfusion of the intestine. Rats that were administered Egb 761 and saline were subjected to laparotomy, ischemia, or ischemia reperfusion procedures. Mucosal lesions were graded from 0 to 5 in histopathological examination. Malondialdehyde and myeloperoxidase levels of the intestinal mucosa were measured. RESULTS: No significant difference was noted between the control and treatment groups regarding the histopathological changes. Although malonyldialdehyde and myeloperoxidase levels of the reperfusion + EGb 761 group were slightly higher than the laparotomy + saline group, they were significantly lower than the reperfusion + saline group. CONCLUSIONS: We concluded that EGb 761 pre-treatment before ischemia-reperfusion decreased malondialdehyde and myeloperoxidase levels and attenuated the mucosal damage. PMID- 11941955 TI - Modified Hassab procedure in the management of bleeding esophageal varices--a two year experience. AB - BACKGROUND/AIMS: Esophageal varices are one of the most common complications (varices, ascites, hypersplenism and encephalopathy) of portal hypertension. Endoscopic sclerotherapy or band ligation is the first choice for esophageal varices today. However, immediate surgical intervention is always needed whenever endoscopic therapy fails to control acute bleeding. Therefore, a simple, effective and less time-consuming procedure is worthwhile for patients who needed immediate surgical treatment. METHODOLOGY: A modified gastroesophageal decongestion and splenectomy GEDS (Hassab) was performed on patients who need immediate surgical intervention for variceal bleeding. There were 15 consecutive patients who received this operative method and the outcome of all patients were reviewed. RESULTS: During follow-up, esophageal varices were resolved in 8 patients (62%), diminished in size in 3 patients (23%) and remain unchanged in 2 patients (15%). Gastric varices disappeared in all patients. The rebleeding rate was 23% in our studies. Encephalopathy was not found in all patients. Moreover, no surgical mortality was found by using this surgical method. CONCLUSIONS: In this study the modified gastroesophageal decongestion and splenectomy (Hassab) was an effective life-saving procedure for its safe, simple and less time consuming advantages. No esophageal transection was performed in this procedure; therefore esophageal fistula, which had high mortality, did not occur. We believe that our procedure may perhaps be an alternative choice for patients who need immediate surgical intervention for bleeding varices. PMID- 11941956 TI - Role of chronic Helicobacter pylori infection in hyperdynamic circulation of cirrhotic patients. AB - BACKGROUND/AIMS: Hyperdynamic circulation observed in portal hypertension is characterized by generalized vasodilatation, increased cardiac index, and increased systemic and regional blood flows, and mediated at least partly by increased nitric oxide activities. Recent studies have demonstrated that Helicobacter pylori (H. pylori) infection can stimulate nitric oxide synthase expression and activities. This study investigated if chronic H. pylori infection might be involved in the development of hyperdynamic circulation in cirrhotic patients. METHODOLOGY: Fifty-eight patients with cirrhosis and thirty-six healthy subjects entered this study. The serologic evidence of H. pylori infection was determined with ELISA in both groups. In addition, in cirrhotic patients hemodynamic studies were performed by Swan-Ganz catheterization and thermodilution technique. RESULTS: No significant differences in age (65.5 +/- 0.8 vs. 63.7 +/- 1.1 years), sex (male/female: 43/15 vs. 29/7) and seroprevalence of H. pylori (74.1% vs. 80.6%) were observed between cirrhotic patients and healthy subjects (P > 0.05). The seropositive rate of H. pylori in patients with cirrhosis was not associated with severity of cirrhosis and size of esophageal varices (P > 0.05). There were no significant differences in systemic vascular resistance and hepatic venous pressure gradient between cirrhotic patients with and those without chronic H. pylori infection (P > 0.05). CONCLUSIONS: The seroprevalence of H. pylori in cirrhotic patients is similar to that of healthy controls, and not related to the severity of cirrhosis and degree of portal hypertension. Chronic H. pylori infection does not play a major role in the hyperdynamic circulation observed in cirrhotic patients. PMID- 11941957 TI - Ultrasound-guided liver resections for hepatocellular carcinoma. AB - Imaging-guided interventional procedures have modified the approach to hepatocellular carcinoma including the surgical one. In fact, liver resections can be carried out with no mortality even if cirrhosis is associated, combining the needs for oncological radicality and liver parenchyma sparing mainly because of the extensive use of intraoperative ultrasonography either for tumor staging or resection-guidance. The aid of intraoperative ultrasonography is therefore optimizing the balance between the oncological radicality and the sparing of the highest amount of functioning liver parenchyma. Intraoperative ultrasonography allows the accomplishment of anatomical resections otherwise not possible such as the systematic segmentectomy. This is of crucial importance if taking into account that anatomical resections seem able to provide better prognosis than the non-anatomical one. However, if non-anatomical resection is carried out intraoperative ultrasonography guidance allows a better tumor clearance. Precise definition of hepatic vein anatomy and association with color Doppler enables hepatectomies otherwise not possible, expanding the indication at surgical resection. In conclusion, we can affirm that liver resection is an imaging-guided procedure and as every interventional imaging-guided procedure, its features are the highest therapeutic efficacy combined with the minimal invasiveness. Then, with the intraoperative ultrasonography guidance liver resection remains the treatment of choice of hepatocellular carcinoma. PMID- 11941958 TI - The Sialyl Lewis X expression in hepatocarcinogenesis: potential predictor for the emergence of hepatocellular carcinoma. AB - BACKGROUND/AIMS: Sialyl Lewis X (sLeX), one of the cancer-associated glycoproteins, has been reported to be expressed in both liver tissue from various types of liver disease and hepatocellular carcinoma. In order to clarify sLeX expression during the early stage of hepatocarcinogenesis, we examined sLeX expressions in either liver tissue specimens without any nodular lesions, dysplastic nodules or hepatocellular carcinomas. METHODOLOGY: Immunohistochemical observations were performed using a monoclonal antibody for sLeX. As for the liver tissue specimens, 8 livers without any chronic liver disease and 42 diseased livers were examined, while for the nodular lesions, 5 dysplastic nodules (borderline lesions) and 47 hepatocellular carcinomas were examined in this study. RESULTS: sLeX was not expressed in all 8 normal livers. sLeX was expressed membraneously in 8 of 15 (53%) chronic hepatitic liver tissue specimens, in 8 of 9 (89%) precirrhotic liver tissue specimens and in 16 of 18 (89%) cirrhotic liver tissue specimens. The incidence of sLeX expression on hepatocytes in both pre-cirrhotic and cirrhotic liver tissue was higher than that in chronic hepatitic liver tissue (P < 0.05). The sLeX expression in liver tissue was positive in all 14 liver tissue specimens containing multiple hepatocellular carcinomas, in which at least one of the nodules was a well-differentiated hepatocellular carcinoma regarded as multicentric hepatocellular carcinomas. In 18 of 28 (64%) liver tissue specimens without multicentric hepatocellular carcinomas, sLeX was positive and the difference was statistically significant (P < 0.05). In nodular lesions, sLeX was negative in 5 dysplastic nodules (borderline lesions). In hepatocellular carcinoma, 14 of 47 (30%) hepatocellular carcinoma nodules showed a positive expression. Six of 14 (43%) well differentiated hepatocellular carcinomas were positive on the cell membrane. Four of 23 (17%) moderately differentiated hepatocellular carcinomas were positive on the cell membrane, while one of 23 (4%) moderately differentiated hepatocellular carcinoma was positive in the cytoplasm. In addition, 3 of 10 (30%) poorly differentiated hepatocellular carcinomas were positive in the cytoplasm. CONCLUSIONS: These results suggested that the sLeX-positive liver tissue specimens possessed a high degree of carcinogenicity and therefore sLeX expression in the diseased liver might be a good predictor for hepatocellular carcinoma emergence. At the same time, the suppression of sLeX occurred at a very early stage of hepatocarcinogenesis. In addition, the phenotype of sLeX was also considered to change during the progression of hepatocarcinogenesis. PMID- 11941959 TI - Effects of streptococcal preparation OK-432 on liver regeneration and energy status after partial hepatectomy under ischemia/reperfusion in rats. AB - BACKGROUND/AIMS: Activation of reticuloendothelial system functions by the treatment with OK-432 has been reported to enhance liver regeneration. However, its effect on liver regeneration has not been studied after hepatectomy under ischemia/reperfusion which is in clinical use. The aim was to examine the effect of OK-432 on regeneration and energy status of the liver after hepatectomy under ischemia/reperfusion in rats. METHODOLOGY: Rats were randomly divided into two groups; OK-432 pretreatment and saline treatment (control) group. In the OK-432 group, OK-432 (2.5 mg/kg body weight) was administered intraperitoneally 24 hours before hepatectomy. In the control group, the same volume of physiological saline was administered in the same manner. Seventy percent hepatectomy was performed in both groups during the second 15-minute ischemia period after an initial 15 minute ischemia and 15-minute reperfusion periods. The survival after hepatectomy, relative liver weight, deoxyribonucleic acid synthesis rate, and hepatic adenine nucleotide and energy charge levels were examined immediately after hepatectomy and on postoperative days 1, 2, 3, and 7. Serum levels of total bilirubin, glutamic pyruvic transaminase, and hyaluronic acid were also measured. RESULTS: All rats survived and the relative liver weight and deoxyribonucleic acid synthesis rate were not significantly different in the two groups. Serum total bilirubin and glutamic pyruvic transaminase levels were not significantly different in both groups. The serum concentration of hyaluronic acid immediately after hepatectomy was significantly higher in the OK-432 group than in the control group. The pretreatment with OK-432 had no significant effect on the levels of adenine nucleotides and energy charge in the liver. CONCLUSIONS: Under ischemia/reperfusion, pretreatment with OK-432 has no significant effect on regeneration and energy status of the liver after hepatectomy. PMID- 11941960 TI - A comparison of right-lobe and left-lobe graft for living-donor liver transplantation. AB - BACKGROUND/AIMS: The practice of living-donor liver transplantation has been widely established. However, in adult cases, the issue of graft liver volume has been raised. Recently, liver transplantation using the right lobe has been done in cases requiring a larger graft volume. We compared right lobectomy with left lobectomy for hepatocellular carcinoma not accompanied by liver cirrhosis and examined the safety of hepatic right lobectomy for donation. METHODOLOGY: In total, 124 hepatocellular carcinoma patients without liver cirrhosis, 89 who had undergone right hepatectomy and 35 who had undergone left hepatectomy, were studied. For each group, we statistically examined the patients' profiles. RESULTS: The resected right lobe was significantly heavier than the left lobe. There were significant differences between the two groups in terms of the number of blood transfusions needed. Moreover, delayed recoveries in terms of hemoglobin, total bilirubin, and clotting disorder on postoperative day 7, as well as the frequency of any complications, were more prevalent in the right-lobe group. CONCLUSIONS: It was confirmed that right lobectomy is more invasive than left lobectomy. It is necessary to prioritize the donor's safety and quality of life in living-donor liver transplantation, and adequate medical and ethical consideration is demanded. PMID- 11941961 TI - Evaluation of total hepatic vascular exclusion and pringle maneuver in liver resection. AB - BACKGROUND/AIMS: The effects of total hepatic vascular exclusion and Pringle maneuver on intraoperative course and postoperative recovery were retrospectively studied. METHODOLOGY: Records of 42 patients who underwent a major hepatectomy and six who had a minor hepatectomy were reviewed. Patients with chronic liver disease or obstructive jaundice were excluded. Hepatic vascular exclusion was used in 5 patients who were at high risk for back flow bleeding from the hepatic veins. Pedicular clamping was used in the other 43 patients. Intergroup differences in intraoperative blood loss, postoperative liver function, the serum interleukin-6 concentrations, and clinical outcome were compared. RESULTS: In four patients of the hepatic vascular exclusion group (n = 5), intraoperative blood loss was less than 2000 mL, and 6000 mL in the remaining patient. However the hepatic vascular exclusion group had longer hepatic ischemia time (45.2 +/- 10.3 min vs. 30.6 +/- 10.9 min), a greater blood loss (2304 +/- 2106 L vs. 913 +/ 1130 mL), a higher serum interleukin-6 concentration (347 +/- 320 pg/mL vs. 93 +/- 58 pg/mL), and a higher morbidity rate (80 vs. 7.1%) compared with the pedicular clamping group (n = 43) (P < 0.05). Postoperative liver function tests were comparable, and no patient developed postoperative hepatic failure. In the pedicular clamping group, intermittent pedicular clamping with periods of 15 minutes (n = 12) increased blood loss (662 +/- 421 mL vs. 1427 +/- 1890 mL), but did not reduce serum interleukin-6 concentration, liver cell damage, or morbidity rate postoperatively, compared with continuous or intermittent clamping of longer periods. CONCLUSIONS: Hepatic vascular exclusion is an effective way to limit blood loss in hepatic resection without causing sever liver injury. However, the interleukin-6 production is increased and the morbidity rate is high. Paradoxically, periodic release of pedicular clamping increases the blood loss but does not reduce liver cell injury or interleukin-6 production. PMID- 11941962 TI - Circulating soluble-CD30 (sCD30) in patients with HCV-related chronic hepatitis and in patients with alcoholic liver disease. AB - BACKGROUND/AIMS: Serum sCD30 (soluble CD30) is a marker of cells producing Th2 type (T-helper-2-type) cytokines. High levels of sCD30 have been found in the active phase of HBV infection. The Th2-type cytokine profile has been documented in alcoholic liver diseases, which have particularly high IgE and IgA serum levels. The aims were: 1) to evaluate sCD30 levels in patients with (a) alcoholic liver diseases and (b) HCV-related chronic hepatitis before and after interferon treatment; 2) to correlate sCD30 concentrations with IgE and IgA serum levels. METHODOLOGY: Serum samples from 34 HCV-related chronic hepatitis patients, before and after interferon treatment, and 17 alcoholic liver disease patients were tested for sCD30 using the ELISA method (Dako, CD30-Ki-1 Antigen, Denmark). RESULTS: Significantly higher levels of sCD30 were found in alcoholic liver disease than in HCV-related chronic hepatitis patients (73.3 +/- 120 vs. 27.5 +/- 44 U/mL, P < 0.05). Alcoholic liver disease patients also exhibited significantly higher levels of IgA than HCV-related chronic hepatitis patients (P < 0.0001). No correlation was found between sCD30 and serum IgA or IgE or response to interferon. CONCLUSIONS: Th2 cells are strongly expanded in alcoholic liver diseases, though the particular immunoglobulin profile observed in this condition has yet to be explained. Th2 function also plays a crucial part in chronic HCV infection, but seems unrelated to interferon response. PMID- 11941963 TI - Early diagnosis of hepatocellular carcinoma using a sensitive assay for serum des gamma-carboxy prothrombin: a prospective study. AB - BACKGROUND/AIMS: Measurement of des-gamma-carboxy prothrombin by conventional methods is of limited use for the early detection of hepatocellular carcinoma for its low sensitivity. The aim of the present study was to investigate the usefulness of measuring des-gamma-carboxy prothrombin by a highly sensitive assay for the early diagnosis of hepatocellular carcinoma in patients with chronic liver disease and for the detection of recurrence after treatment of hepatocellular carcinoma. METHODOLOGY: Des-gamma-carboxy prothrombin levels by a sensitive assay and alpha-fetoprotein levels were sequentially measured in 188 patients with type B or C chronic liver disease and in 63 patients with hepatocellular carcinoma. RESULTS: The positive rate of des-gamma-carboxy prothrombin was 62% in all of hepatocellular carcinoma patients. Hepatocellular carcinoma was detected in 14 of 188 chronic liver disease patients during their follow-up period, the positive rate of des-gamma-carboxy prothrombin and of alpha fetoprotein being 57% and 71% in these 14 patients, respectively. Des-gamma carboxy prothrombin level normalized in 67% of 39 patients after the treatment of hepatocellular carcinoma. Of the 19 patients with tumor recurrence, 84% showed re elevation of des-gamma-carboxy prothrombin level. CONCLUSIONS: Measurement of des gamma-carboxy prothrombin by this highly sensitive assay combined with alpha fetoprotein is useful for detecting hepatocellular carcinoma in chronic liver disease patients and for monitoring recurrence after treatment of hepatocellular carcinoma. PMID- 11941964 TI - Assessment of histological features in chronic hepatitis C. AB - BACKGROUND/AIMS: Hepatitis C virus infection is an important disease with a high chronicity rate (50-80%), leading to end-stage liver cirrhosis and hepatocellular carcinoma. In this study, the characteristic histological findings were compared with the epidemiological features of hepatitis C virus infection in liver biopsy material. METHODOLOGY: Liver biopsies were studied from 106 patients (60 males, 46 females, aged 11-81 years, mean: 43) found positive for hepatitis C antibody by a second-generation ELISA method. The histological evaluation was based upon the Desmet classification of chronic hepatitis. Statistical analysis involved the chi 2 test. RESULTS: Minimal, mild chronic, and moderate chronic hepatitis were manifest in 13%, 65% and 22% of the cases, while fibrosis 0/1, fibrosis 2, fibrosis 3 and cirrhosis (fibrosis 4) occurred in 42%, 13%, 35% and 10%, respectively. Regarding the histological features of chronic hepatitis C, the frequency of steatosis, lymphoid follicles and/or aggregates and bile duct lesions showed an increase parallel with the hepatitis grade, the changes being more pronounced in moderate chronic hepatitis. CONCLUSIONS: Most chronic hepatitis C patients displayed mild histological lesions with stage 1 fibrosis. Lymphoid follicles and/or aggregates, bile duct damage and steatosis are important diagnostic features that show strong correlation with chronic hepatitis activity. The occurrence of stage 3/4 fibrosis in mild chronic hepatitis alerts the hepatologists for the need of liver biopsy. PMID- 11941965 TI - Esophagogastric devascularization as the last option in the management of variceal bleeding. AB - Variceal bleeding is a clinical emergency that may be difficult to treat in some patients, especially those with prehepatic portal hypertension, failed sclerotherapy, and with contraindications to transjugular intrahepatic portosystemic shunt. In such patients there are few remaining options. The authors refer to three patients for whom the modified Sugiura procedure was the only remaining option for the treatment of variceal bleeding. Two of them had pre hepatic portal hypertension, and one had hepatic cirrhosis, and in all, other standard treatment options and failed. The modified Sugiura devascularization and esophageal transection was performed without operative or postoperative complications and in the follow-up, (mean: 4.2 years), there was no recurrence of variceal bleeding. The authors recommend the modified Sugiura procedure as safe and effective for patients in whom other treatment options for variceal bleeding have failed. PMID- 11941966 TI - Hepatectomy for liver metastasis from ampullary cancer after pancreatoduodenectomy. AB - A 51-year-old Japanese woman with a solitary liver metastasis originating from a carcinoma of the ampulla of Vater was successfully treated by partial hepatectomy 19 months after curative pancreatoduodenectomy with lymphadenectomy. Histologic examination revealed a stage III well-differentiated tubular adenocarcinoma (pT2, pN1, and pM0). Postoperative serum concentrations of carcinoembryonic antigen increased exponentially to 133 ng/mL. The carcinoembryonic antigen doubling time was 63 days. Computed tomography and ultrasonography of the abdomen showed a solitary metastasis in segment VI of the liver. Since neither local recurrences nor other distant metastases were detected, the patient underwent partial hepatectomy. Histologic study confirmed the presence of a metastatic liver tumor from the ampullary carcinoma. The carcinoembryonic antigen levels returned to normal immediately after the partial hepatectomy. She was well without signs of recurrence 18 months after partial hepatectomy. PMID- 11941967 TI - Bilateral adrenal metastases from hepatocellular carcinoma after liver transplantation. AB - Therapeutic management of hepatocellular carcinoma is a controversial issue. Orthotopic liver transplantation is an alternative for the treatment of hepatocellular carcinoma in a selected group of patients, but recurrence is possible. A 51-year-old patient with liver transplantation due to hepatocellular carcinoma presented bilateral adrenal metastases in a successive manner. A left adrenal gland metastasis was diagnosed five months after liver transplantation, and a left adrenalectomy was carried out. Eight months later, a right adrenal gland metastasis was diagnosed, and a right adrenalectomy was performed. Pathological examination confirmed the diagnosis of a well-differentiated hepatocellular carcinoma. At present, there is no evidence of recurrence 35 months after the second adrenalectomy. Bilateral adrenal gland metastases from hepatocellular carcinoma after liver transplantation have not been previously reported in English literature. Surgical resection of metastases may be indicated in similar patients with successful treatment of the primary tumor, absence of additional metastasic disease, and good performance status. PMID- 11941968 TI - Metronidazol as a probable cause of severe liver injury. AB - Metronidazol, a commonly used antibiotic drug, has been very rarely associated with hepatotoxicity. In particular, no reports have appeared in the literature about cases of metronidazol-associated severe hepatotoxicity, leading to liver transplantation or death. We report on a case of acute fulminant liver failure in a young woman, who had, two years previously, developed jaundice after intake of metronidazol. During the current hospitalization, metronidazol treatment had been undertaken two weeks previously and also this time the patient developed severe hepatocellular injury and cholestasis. A viral etiology was ruled out as well as vascular, metabolic and malignant etiology. Although, the cause of the liver injury in this case is not proven, the relationship between this drug and two occasions of severe liver damage, suggests a positive challenge as well as rechallenge. An International algorithm was used for the assessment of the causality of a drug in this case of acute liver injury and a "probable" classification was obtained. PMID- 11941969 TI - New reconstructive surgery of remnant pancreas in cases of cancer of Vater's papilla. AB - BACKGROUND/AIMS: The radical surgical procedure for treatment of the carcinoma of papilla of Vater is the pancreatoduodenectomy. The mortality rate of the surgery highly decreased in the last decade, nevertheless there are complications related to the complication of anastomosis of the remnant pancreas. METHODOLOGY: The authors introduce a new reconstructional procedure to decrease the complications. After the removal of the pancreatic head and body an end-to-side anastomosis was performed between the pancreatic duct and a Roux-en jejunal loop. The second anastomosis of the procedure was an end-to-side choledochojejunostomy, the third was an end-to-side duodenojejunostomy. The duodenojejunostomy is about 40 cm from the pancreatic anastomosis, keeping food far from the pancreas with the help of peristaltic waves. This method was applied in 6 patients. RESULTS: It was found that the new reconstructional procedure had generally favorable results without complication. CONCLUSIONS: This method of reconstruction allows for spontaneous closure and safe drainage of potential insufficient pancreaticojejunostomy. The recovered patients support future favorable usage of this new reconstructional surgical procedure. PMID- 11941970 TI - Arterial and vena caval resections combined with pancreaticoduodenectomy in highly selected patients with periampullary malignancies. AB - BACKGROUND/AIMS: To obtain a margin-negative resection and increase the indication for resection of periampullary malignancies, pancreaticoduodenectomy with a SM-PVR (superior mesenterico-portal vein resection) has been performed. However, an arterial resection, other vascular resections except SM-PVR (e.g., an inferior vena caval resection), or a metastatic tumor resection combined with pancreaticoduodenectomy has yet to be fully elucidated because of the high risk of postoperative complications and extremely poor long-term survival in patients undergoing these exceptional procedures. The present report focused on highly selected patients undergoing an arterial resection or a vena caval resection associated with pancreaticoduodenectomy. METHODOLOGY: Besides 31 patients with periampullary tumors undergoing pancreaticoduodenectomy associated with SM-PVR in our department, a group of 4 patients underwent arterial resections and another patient underwent pancreaticoduodenectomy combined with a resection of liver metastasis together with an inferior vena caval resection. These five patients were reported in the present study. RESULTS: A 27 year-old-woman presented pancreatic ductal adenocarcinoma of the pancreatic head and a liver metastasis in which involvements of the superior mesenterico-portal vein and the inferior vena cava were shown. Pancreaticoduodenectomy was performed with SM-PVR associated with a left hemihepatectomy combined with a segment 1 resection and an inferior vena caval resection. The patient did not present severe postoperative complications and experienced a good quality of life during 16 months after surgery. Four other patients underwent arterial resections. These arterial resections were performed only when a margin-negative resection was feasible. The superior mesenteric artery was resected and reconstructed with a Goretex graft in one patient. The right hepatic artery was resected and reconstructed with a saphenous graft in two patients. The other patient underwent a resection of the common hepatic artery and reconstruction was performed with the splenic artery. Three of the four patients presented postoperative complications but were conservatively treated. Two patients are still alive 25 months and 8 months after surgery. One patient died of sepsis 5 months after surgery, and the other died of cancer progression 19 months after surgery. CONCLUSIONS: The indication for retropancreatic arterial resection associated with pancreaticoduodenectomy should be carefully evaluated only when a margin-negative resection can be achieved. An appropriate bypass method of arterial reconstruction should be selected because a direct end-to-end anastomosis is not always feasible. Hepatectomy for metastases of pancreatic ductal carcinoma should be also regarded as an exceptional procedure. PMID- 11941971 TI - Clinicopathologic features and outcomes of intraductal papillary-mucinous tumors of the pancreas. AB - BACKGROUND/AIMS: The authors reviewed their experience with resected IPMT (intraductal papillary-mucinous tumor) of the pancreas to clarify the characteristics and prognosis of this neoplasm. METHODOLOGY: Between 1983 and 1998, 30 consecutive patients with IPMT underwent operations at our institution. Their clinicopathological features and postoperative long-term outcomes were analyzed retrospectively. RESULTS: There were 22 males and 8 females, with a mean age of 64 years. Operations performed were duodenum-preserving pancreatic head resection in 12 patients, distal pancreatectomy in 8, segmental pancreatectomy in 6, conventional pancreaticoduodenectomy in 4. Malignancy was found in 10 of 30 (33%). Factors significantly associated with malignancy were tumor size or presence in main pancreatic duct. In 30 resected patients after a mean follow-up of 60 months, tumor recurrence had occurred in 2 cases of invasive carcinoma that infiltrated into the extrapancreatic organ. The overall actuarial 5-year and 10 year survival was 83% and 62%, respectively. CONCLUSIONS: IPMT has a favorable prognosis after adequate resection. Despite slow growth, IPMT has an obvious malignant potential and a poor prognosis when invasive carcinoma has developed. Early recognition and pancreatectomy is the mainstay of treatment for IPMT. PMID- 11941972 TI - Pancreatic carcinoma-induced changes in gastric myoelectric activity and emptying. AB - BACKGROUND/AIMS: Twenty patients with histologically confirmed pancreatic carcinoma without any endoscopic evidence of gastroduodenal obstruction were included in the study. The aim was to determine changes in gastric myoelectric activity and liquid/solid gastric emptying induced by pancreatic tumor. METHODOLOGY: According to TNM/UICC classification patients were divided into two groups A (T2) and B (T3) due to extent of tumor invasion (mainly to retroperitoneum space). In all patients electrogastrography, solid and liquid gastric emptying tests were performed. RESULTS: In the majority of patients of groups A and B the most commonly reported complaints included upper abdominal pain (60% vs. 80%) and icterus (80% vs. 60%). Dyspeptic symptoms were observed in 40% patients of group A and 90% in group B. In group electrogastrography recordings showed dysrhythmia patterns, mostly bradygastria, in 50% of group A patients and in 80% of group B. Liquid/solid gastric emptying were delayed in 20/40% of group A patients and 50/80% of group B. Disorders of gastric myoelectric activity and emptying correlated with tumor stage and location across analyzed groups but not with histology and hyperbilirubinemia levels. CONCLUSIONS: It was observed that solid gastric emptying is affected earlier compared to liquid gastric emptying. Delayed gastric emptying may be attributed to gastric dysrhythmia and/or abdominal pain but not mechanical effects of tumor growth that occur during the course of disease. PMID- 11941973 TI - CT-guided drainage of a mediastinal pancreatic pseudocyst with a transhepatic transdiaphragmatic approach. AB - We describe a mediastinal pancreatic pseudocyst treated with a catheter drainage placed under computed tomographic guidance using a newly applied approach. A needle was angled cephalad with the computed tomographic gantry tilt technique, and was inserted into the lowest portion of the cavity through the liver and through the diaphragm. The needle pathway was necessitated by the location of the pseudocyst cavity. The drainage alleviated completely the symptoms with no complications. The puncture technique should be an option for accessing mediastinal lesions percutaneously. PMID- 11941975 TI - The effect of prostacyclin on the gastric mucosa in portal-hypertensive rats. AB - BACKGROUND/AIMS: This study investigates the effect of prostacyclin, which is thought to be involved in the hemodynamic circulation, on the gastric mucosa of rats with portal hypertension. METHODOLOGY: Various gastric functions were evaluated in portal vein ligated rats after the intraperitoneal administration of either a placebo or prostacyclin for 7 days. RESULTS: The gastric mucosal damage induced by the instillation of 90% ethanol was significantly greater in the prostacyclin-treated group than in placebo-treated group. The portal pressure was similar in both groups. There was no significant difference between the two groups in plasma concentration of 6-keto-PGF1a (a stable metabolite of prostacyclin), whereas the mucosal content of 6-keto-PGF1a was significantly higher in the prostacyclin-treated group than in the placebo-treated group. Prostacyclin pretreatment significantly increased the gastric mucosal blood flow, estimated by laser-Doppler flowmetry, and the hemoglobin content of the gastric mucosa, measured by reflectance spectrophotometry, whereas the oxygen content remained unchanged. CONCLUSIONS: We speculate that the increased gastric mucosal perfusion induced by a high content of prostacyclin in the portal hypertensive gastric mucosa may play a role in the pathogenesis of ethanol-induced gastric mucosal damage. PMID- 11941976 TI - The vascular control in liver resection: revisitation of a controversial issue. AB - Limiting intraoperative blood loss and the consequent need for whole blood transfusion is a widely accepted goal in liver resection. To achieve this goal, liver resection carried out under warm ischemia seems adequate. Methods used for this purpose can be categorized as follows: those with only an inflow control and the one that consists of total vascular exclusion. Liver resections under inflow vascular control are safer than those performed without. Furthermore, up to now the ischemia-reperfusion liver damage does not seem to affect the patients' course. The clinical evidence shows that intermittent warm ischemia seems to be safer than the continuous clamping and guarantees an effective control of the intraoperative bleeding. Conversely, total vascular exclusion is an invasive technique with not negligible morbidity; then its real indications should be restricted to exceptional cases, such as those with infiltration of the inferior vena cava which demands substitution of the involved vessel. In conclusion, up to now intermittent warm ischemia is the most appropriate approach to carry out safe liver resections. PMID- 11941974 TI - Pancreatic pseudocystportal vein fistula manifests as residivating oligoarthritis, subcutaneous, bursal and osseal necrosis: a case report and review of literature. AB - Pseudocyst is a common complication of pancreatitis. Pseudocyst may rupture into the surrounding organs. Rupture into the portomesenteric vein is extremely rare with only seven cases being described in the English literature. pancreatic portal vein fistula is very difficult to verify. The aim of this study was to view the diagnostic methods of pancreatic portal vein fistula and to describe the results of high-dose corticosteroid treatment to our knowledge for the first time. We report here a case of pancreatic portomesenteric vein fistula that was manifest as subcutaneous fat necrosis, bursal necrosis, intramedullary aseptic bone necrosis and recidivating oligoarthritis. The literature of this unusual complication is reviewed. The results of high-dose corticosteroid treatment are also described. In patients with recidivating oligoarthritis, subcutaneous, bursal or osseal necrosis a pancreatic process should be included in the differential diagnosis even in cases of no abdominal signs or symptoms or previous abdominal history. Operative exploration of the pancreas should be performed in the early phase of the disease. To diminish the ongoing extrapancreatic manifestations after the closure of the fistula massive corticosteroid treatment may be attempted although the role of this therapy remains controversial. PMID- 11941977 TI - Alterations of BAT-26 identify a subset of gastric cancer with distinct clinicopathologic features and better postoperative prognosis. AB - BACKGROUND/AIMS: Gastrointestinal tumors with microsatellite instability represent a replication error-positive phenotype. BAT-26, a repeat of 26 deoxyadenosine localized in intron 5 of hMSH2 gene, has been reported as a reliable indicator of replication error phenotype in colorectal cancers. This study investigated whether BAT-26 is a useful marker for a mutator phenotype with distinct clinicopathologic features in gastric cancer. METHODOLOGY: One hundred and nineteen gastric cancer tissues and matched non-tumor tissue were examined by polymerase chain reactions with electrophoresis for 9 dinucleotide microsatellites and BAT-26, and frameshift mutations of transforming growth factor-beta type II receptor. The relation between BAT-26 alterations and relevant clinicopathological or genetic parameters were analyzed. RESULTS: Gastric cancer with BAT-26 alterations was highly correlated with multiple microsatellite alterations (> or = 3 loci) and frameshift mutations of transforming growth factor-beta type II receptor, and predominantly showed antral location, intestinal histologic subtype, advanced stage, a higher rate of Helicobacter pylori infection, a better postoperative survival and less lymph node metastasis. CONCLUSIONS: These results show testing of BAT-26 alterations is a convenient and rapid screening method for identifying a subset of gastric cancer with a mutator phenotype and better prognosis. PMID- 11941979 TI - Limits and benefits of exclusive transthoracic hepatectomy approach for patients with hepatocellular carcinoma. AB - BACKGROUND/AIMS: The purpose of this study was to evaluate the results of liver resection in cirrhotic patients for liver hepatocellular carcinoma located near the diaphragm through an exclusive transthoracic approach. METHODOLOGY: Between 1995 and 1999, 19 cirrhotic patients with hepatocellular carcinoma underwent a liver resection through an exclusive transthoracic approach. This approach was indicated in 11 cases for previous upper abdominal surgery, including hepatobiliary surgery in 3 and before liver transplantation in 8. Results of the transthoracic approach were compared to 84 cirrhotic patients who underwent transabdominal limited resection of hepatocellular carcinoma matched for age, sex and localization of the tumor. RESULTS: Resection was feasible by an exclusive transthoracic approach in 18 (95%) cases with a mean operating time of 201 +/- 53 min. In 8 (44%) patients a Pringle maneuver was performed. No postoperative deaths were observed after the transthoracic approach. Pulmonary complications rate was significantly higher (P < 0.001) after transthoracic resection compared to transabdominal resection (67% vs. 25%, P < 0.001). In contrast, ascites were observed in only one (5%) of the transthoracic group compared to 35 (42%) in the transabdominal group (P < 0.01). The resection margin was positive in 3 (17%) after transthoracic approch and in 1 (2%) patient after the transabdominal resection (P < 0.02). In patients who underwent liver transplantation after the transthoracic approach, total hepatectomy was performed without increasing difficulties. CONCLUSIONS: The transthoracic approach is a safe procedure for resection of hepatocellular carcinoma located under the right diaphragm in cirrhotic patients. However, this approach allows only limited resection with a high risk of positive margin, resulting in a restriction of indications either for patients with previous major abdominal surgery than before liver transplantation. PMID- 11941978 TI - Gastric mucosal exposure to histamine in rats is followed by absorption of histamine but not mucosal hyperemia. AB - BACKGROUND/AIMS: Superficial mucosal damage is associated with mucosal hyperemia and release of histamine into venous blood and gastric lumen. This study was designed to examine if histamine administrated directly to gastric mucosa becomes absorbed and transported away with blood, and if so whether it is capable of increasing gastric mucosal blood flow. METHODOLOGY: Superficial mucosal injury was induced by exposing chambered rat stomachs to 2 M NaCl for 10 min, and 45 min later 4.5 x 10(-2) or 4.5 x 10(-3) M histamine was instilled into the chamber. Mucosal blood flow was determined by radioactive microspheres and continuously by Laser Doppler flowmetry. RESULTS: Histamine concentration in arterial blood was markedly increased 15 min after administration of histamine. The mucosal blood flow did not increase after application of histamine. Significant increase in heart rate and fall in arterial blood pressure were observed. Similar results were obtained after application of histamine to normal mucosa. CONCLUSIONS: Histamine applied to injured or normal gastric mucosa of rats becomes transported via capillaries and venules to the systemic circulation, but does not reach terminal arterioles in quantities large enough to cause vasodilatation. PMID- 11941980 TI - Remodeling the surgical approach to hepatocellular carcinoma. AB - Surgery for hepatocellular carcinoma has improved during the last two decades, and the improvement is mainly attributable to various innovations in liver surgery, such as establishment of the precise criteria for surgical indications, development of ultrasound-guided hepatectomy, and additional use of portal vein embolization. Operative mortality has fallen below 2% in the 1990's, and the 5 year survival rate reached, according to the results of a nationwide survey, nearly 50%. More than 90% of the hepatectomies in the authors' institution are performed without whole blood transfusion, and mean hospital stay is approximately 23 days. Moreover, no-mortality hepatectomy has been achieved since 1993. PMID- 11941981 TI - Resection of hepatocellular carcinoma: a European experience on 328 cases. AB - BACKGROUND/AIMS: Surgical liver resection has been demonstrated in Asian countries to be the best therapeutic option in patients with hepatocellular carcinoma. Because the value of this treatment is still debated in Western countries, the aim of this paper was to report a European experience of resection for hepatocellular carcinoma. METHODOLOGY: From 1990 to 1999, 239 men and 61 women aged from 15 to 77 years old underwent 328 resections including major resection in 138 (42%) cases. Normal liver was present in 53 patients (17%) and chronic liver disease was present in 247 including 152 (50%) with cirrhosis. RESULTS: In-hospital mortality was 6.4% and was significantly influenced by the presence of chronic liver disease (1.7% vs. 7.4%). Mortality after resection in alcoholic patients (14%), in patients with hepatitis C (9%) was significantly higher than in patients chronic hepatitis B (1%) (P < 0.05). The overall survival rates were 81%, 57%, 37%, and 13% at 1, 3, 5 and 10 years. Five-year survival rate was significantly higher (P < 0.05) in patients with normal liver as compared to chronic liver disease (50% vs. 34%). In patients with chronic liver disease parameters, which significantly influenced survival rate, were vascular invasion, tumor differentiation and the extent of resection. CONCLUSIONS: In this European study with varied profile of etiologies associated with hepatocellular carcinoma we showed that a five-year survival rate of 40% can be expected after resection and that chronic liver disease is a major factor influencing short and long-term prognosis. PMID- 11941982 TI - Liver transplantation for hepatocellular carcinoma. AB - Liver transplantation has become the best option in patients with decompensated cirrhosis and a small hepatocellular carcinoma. Indeed, because of the severity of cirrhosis, resection is usually impossible and in addition, transplantation provides survival rates close to those obtained in cirrhotic patients without malignancy (70 to 80% 3-year survival rate). In patients with a small hepatocellular carcinoma and compensated cirrhosis, both resection and transplantation can be performed. Because of the scarcity of donors, there have been reservations concerning transplantation in patients who otherwise could have undergone resection. However, there is increasing evidence that long-term results of transplantation are significantly superior to those of resection. Therefore, patients with a small hepatocellular carcinoma and compensated cirrhosis are increasingly considered as suitable candidates for transplantation. In contrast to cirrhotic patients with a small hepatocellular carcinoma, patients with large and/or multifocal tumors should no longer be transplanted because of a high rate of early recurrence and the accelerated course of tumor progression due to immunosuppression, both factors being the source of poor results. On rare occasions, hepatocellular carcinoma develops in patients without underlying liver disease. In such cases the tumor is usually recognized when it is large and symptomatic. The absence of underlying liver lesions offers the possibility of extended resection. However, in case of nonresectable (bilobar) tumors or limited recurrence after resection, transplantation may be considered due to the slow progression this subtype of hepatocellular carcinoma. Whatever the underlying liver parenchymal status, efforts should be made to reduce the risk of recurrence. PMID- 11941983 TI - Reasons for an exchange between eastern and western approach to patients with HCC. PMID- 11941984 TI - Living-donor liver transplantation for hepatocellular carcinoma. AB - The current status of cadaveric liver transplantation in Japan and our experience with living-donor liver transplantation for patients with hepatocellular carcinoma are described. At Shinshu University, we performed 137 living-donor liver transplantations on 137 patients, including 80 pediatric and 57 adult patients, between June 1990 and November 2000. The survival rates for these patients are 87% at 1 year and 84% at 5 years. Of these 137, eight had hepatocellular carcinoma. Only one patient had a solitary tumor while in the remaining seven patients, the tumors were multiple, ranging in number from 2 to 11. The maximum tumor size was within the range 14 to 45 mm, and six out of the eight were Stage III or IVa. With a follow-up ranging from 2 to 119 months, two patients died, one patient had recurrence while the remaining are all alive and disease-free. These data, due to the relatively short follow-up and the small number of cases, do not allow us to draw any valuable conclusions about the effectiveness of living-donor liver transplantation in the treatment of hepatocellular carcinoma. However, in consideration of the worldwide recognized good therapeutic efficacy of liver transplant from cadaveric donors in the treatment of hepatocellular carcinoma and also the relatively advanced stage of the disease in the present series, these preliminary results seem promising. PMID- 11941985 TI - Perspectives and drawbacks of minimally invasive surgery for hepatocellular carcinoma. AB - The need for an accurate intrahepatic staging is crucial for patients with hepatocellular carcinoma candidates to an aggressive surgical or ablative treatment. Currently available data indicate that laparoscopy with laparoscopic ultrasound provides information similar to that obtained by intraoperative ultrasound and it is able to identify small intrahepatic lesions not diagnosed by preoperative imaging techniques. Furthermore, laparoscopy with laparoscopic ultrasound also allows performance of ultrasound-guided biopsies or interstitial therapies as ethanol injection, cryoablation or radiofrequency thermal ablation in the same session. A laparoscopic segmentectomy or subsegmentectomy is technically feasible and safe in selected patients with small peripheral tumors. Combinations of resection and ablation may be required in certain cases, extending the indications for the laparoscopic approach to hepatocellular carcinoma in liver cirrhosis. The AA review the technical issues and the preliminary results of their experience in the field of minimally invasive approach to hepatocellular carcinoma. On the basis of these preliminary findings, laparoscopy with laparoscopic ultrasound seems to be useful to identify unsuspected new nodules and to help in choosing the most suitable treatment. In case of hepatocellular carcinoma not amenable to surgical resection, laparoscopic radiofrequency represents a safe and effective treatment above all when the percutaneous approach is difficult or impossible. Furthermore, laparoscopy with laparoscopic ultrasound could represent a sound preliminary examination in patients who are candidates to liver transplantation in order both to improve the staging and to guide an interstitial therapy as a bridge to the transplantation itself. PMID- 11941986 TI - Treatment of hepatocellular carcinoma by percutaneous interventional methods. AB - In the treatment of early and intermediate hepatocellular carcinoma the range of indications for percutaneous ablation techniques is becoming wider than surgery or intra-arterial therapies. Indeed, whereas for some years only patients with up to three small tumors were treated, with the introduction of the single-session technique performed under general anesthesia, even patients with more advanced disease are now being treated. Although it is understood that partial resection assures the highest local control, the survival rates after surgery are roughly comparable with percutaneous ethanol injection. The explanation is due to a balance among advantages and disadvantages of the two therapies. Percutaneous ethanol injection survival curves are better than curves of resected patients who present adverse prognostic factors, and this means that surgery needs a better selection of the patients. Indications for both of these therapies are reported. An open question remains about the choice between percutaneous ethanol injection and other new ablation procedures. In our department we currently use radiofrequency ablation in the majority of patients but consider percutaneous ethanol injection and segmental transarterial chemoembolization complementary, and use them according to the features of the disease and the response. Evaluation of their therapeutic efficacy, techniques and results are reported. PMID- 11941987 TI - Prognosis of hepatocellular carcinoma. AB - The prognosis of patients with hepatocellular carcinoma is related to the stage of the tumor at diagnosis and to the degree of liver function impairment induced either by the tumor itself or by the underlying cirrhosis. Any prognostic prediction should also take into account the potential impact of therapeutic interventions. Only surgical resection, liver transplantation and percutaneous ablation achieve a relatively high rate of complete responses in patients with tumors diagnosed at an early stage and may improve survival. By contrast, patients diagnosed at an advanced stage will receive palliative treatment with unproven survival benefits. Each stage and each treatment have their specific prognostic predictors. Thus, the most accurate prognostic system will have to use a specific model for each strata at which patients may be diagnosed: early, intermediate-advanced and terminal. Patients at an early stage may achieve a 5 year survival rate above 50%, those at intermediate-advanced present a 20-50% survival at 3 years and those at terminal stage die within six months. In addition to predicting prognosis, the staging system should also guide the selection of treatment and this is the major advantage of the classification applied in the Barcelona-Clinic Liver Cancer Group. PMID- 11941988 TI - Transcatheter arterial chemoembolization of hepatocellular carcinoma: a Japanese experience. AB - Hepatocellular carcinoma is one of the most common causes of cancer death in Japan and in 80% of cases is associated with chronic liver disease caused by hepatitis C virus. Poor hepatic function reserve due to underlying cirrhosis is the primary factor which limits extended surgical resection in many cases. Furthermore, in patients treated by curative resection, high incidence of recurrent tumors or/and newly developed tumor in the residual liver was reported. Therefore, the aim of various therapeutic options such as operation, percutaneous ethanol injection, radiofrequency coagulation therapy and transcatheter arterial chemoembolization should be the local control of hepatocellular carcinoma. Transcatheter hepatic arterial chemoembolization has a main role for the multidisciplinary treatment for hepatocellular carcinoma with this biological behavior. PMID- 11941989 TI - Hepatic artery chemoembolization for advanced stage HCC: experience of 650 patients. AB - Hepatic artery chemotherapy using cisplatin in various protocols was examined in 650 patients. Overall objective tumor response rate (PR) was 65%. Average survival was 7.5 mo in patients with tumor progression, 18.0 mo for tumor stability and 32.0 mo for PR. 1- and 2-yr survival was 70% and 40% in responders, 20% and 0% in progressors. Prognostic factors were examined in 155 patients treated with cisplatin and gelfoam chemo-occlusion. In survival groups of > 24 mo, 4-24 mo and < 4 mo, similar numbers had cirrhosis, hepatitis B virus, hepatitis C virus and alcoholism. Decreased survival was associated with abnormal bilirubin, albumin and prothrombin time. Tumor vascularity and response to chemotherapy were associated with prolonged survival. Tumor vascularity seemed important for tumor response. Portal vein thrombosis occurred in all groups. Lesion number, bilobarity and maximum size had no correlation with response or survival. We analyzed the cause of death in 425 patients. No evidence of hepatocellular carcinoma progression, judged by absence of change in CT scan or tumor marker in the last 4 months of life, was found in 42%. A group of 57 patients were treated with cisplatin in dose range 125-200 mg/m2 alone or with gelfoam. In both groups, responders survived longer than non-responders: cisplatin alone responder mean survival, 29.0 mo, non-responder 11.1 mo, P < 0.0001. There was a strong effect of dose density on median survival for cisplatin alone, but not for cisplatin and gelfoam. CONCLUSIONS: A large experience of single-agent cisplatin chemo-occlusion is summarized. Good liver function and tumor vascularity are associated with response to chemotherapy, which in turn is associated with enhanced survival. Many deaths are due to cirrhosis and not hepatocellular carcinoma. PMID- 11941990 TI - Prevention of hepatocellular carcinoma recurrence: actuality and perspectives. AB - Postsurgical recurrence of hepatocellular carcinoma is frequent and fatal. Various adjuvant treatments to possibly prevent recurrence have been recommended, which seem to depend on expectation rather than evidence. Up to now, randomized controlled trials have been conducted to clarify the clinical effect of the 4 therapeutic options including acyclic retinoid, 131I-lipiodol, adoptive immunotherapy, and interferon. We have recently demonstrated that adoptive immunotherapy is a safe, feasible treatment that can reduce the risk of recurrence and improve recurrence-free outcomes. Actually all the options reduced in part the recurrence but had drawbacks in the their effectiveness, and large trials are needed to assess other important endpoints, such as clinical feasibility, risk-benefit and cost-effectiveness. Recurrence control of hepatocellular carcinoma is the clinical priority, and we are approaching this goal. PMID- 11941991 TI - Development and perspectives of bioartificial liver support. AB - Bioartificial liver support systems containing adsorbent devices, xenogeneic whole liver perfusion, and hybrid bioartificial liver are anticipated to be effective for the treatment of severe hepatic failure. At present, whole liver perfusion and the hybrid bioartificial liver are two mainstreams in the field of the bioartificial liver, but it is still unclear whether either of them has significant beneficial effects in hepatic failure patients. We developed a new system of xenogeneic direct hemoperfusion consisting of a leukocyte adsorbent, an immunoglobulin adsorbent, and a substitute unit for hepatic function, that is, whole liver or bioreactor. Using this system, the perfusion efficiency rate and oxygen supply of the substitute unit for hepatic function were significantly increased. A further improvement of our system by incorporating a leukocytapheresis device to return the leukocyte-rich portion of the perfused blood back to the patient's body directly was undergone. Moreover, our system can be applied not only as a bioartificial liver, but also as a bioartificial kidney. To solve the problem of xenogeneic protein influx into the human body associated with extracorporeal bioartificial liver support, it is important to develop transgenic cattle in which human protein genes are transduced. Influx of porcine endogenous retrovirus into the human body has been a controversial subject. In relation to this issue, the cross-hemoperfusion method is promising in that the patient's circuit and the bioartificial liver circuit are separated by a semipermeable membrane, which can prevent any kind of virus from flowing into the patient's circuit. PMID- 11941992 TI - Cholangiocellular carcinoma depending on the kind of intrahepatic calculi in patients with hepatolithiasis. AB - BACKGROUND/AIMS: Association of cholangiocellular carcinoma in patients with hepatolithiasis has been reported. However, its incidence depending on the kind of stones is obscure. The aim was to examine the association rate of cholangiocellular carcinoma in patients with hepatolithiasis with a special reference to the kind of intrahepatic stones. METHODOLOGY: One hundred and thirty nine patients with hepatolithiasis who have been treated from 1973 to 1997 were retrospectively reviewed to examine the characteristics of cholangiocellular carcinoma. The type and location of intrahepatic calculi were analyzed. RESULTS: Cholangiocellular carcinoma was found in 8 of 139 patients, the incidence being 5.8%. The incidence of carcinoma was more than twice in patients whose stones were located in intrahepatic bile duct only (9.3%; 5/54) than in those located in both intrahepatic and extrahepatic bile duct (3.5%; 3/85). Of 121 patients excluding 18 whose calculi were not available for classification, the kind of intrahepatic calculi was brown pigment in 106 patients (87.6%), cholesterol in 10 (8.3%), black pigment in 4, and fatty acid calcium in one. Cholangiocellular carcinoma was associated in 3 (2.8%) of 106 patients with brown pigment stones, 3 (30%) of 10 with cholesterol stones, in one with fatty acid calcium stones, and one patient whose stone was not available for analysis. The association rate of cancer was significantly (P < 0.01) higher in patients with cholesterol stones than those with brown pigment stones. One patient survived for 24 months after left lobectomy but the others died within six months. CONCLUSIONS: An early and attentive evaluation for the possible presence of cholangiocellular carcinoma is mandatory not only in patients with brown pigment stones but also in those with intrahepatic cholesterol stones. PMID- 11941993 TI - Why do multi-organizational quality initiatives usually fail? PMID- 11941994 TI - Lack of relationship between the Cleveland Health Quality Choice project and decreased inpatient mortality in Cleveland. AB - The Cleveland Health Quality Choice (CHQC) project was previously suggested to have been responsible for declines in inpatient mortality in Cleveland hospitals during the first years of the project (1992-1993). We sought to (a) verify this decline in mortality and (b) better understand its possible relationship to CHQC. We employed regression analysis to compare mortality rate trends for Cleveland versus the rest of Ohio using Ohio Hospital Association inpatient mortality data. We found that the rate of decline in mortality in Cleveland (-.218% per 6 months; 95% confidence limits, -.278% to -.159% per 6 months) was statistically indistinguishable from that in the rest of the state (-.188% per 6 months; 95% confidence limits, -.234% to -.143% per 6 months) (P = .35). We could not demonstrate a significant beneficial effect of the CHQC project on hospital mortality in Cleveland. Inpatient mortality was simultaneously declining at a statistically indistinguishable rate throughout the state. PMID- 11941995 TI - The autopsy: a professional responsibility in assuring quality of care. AB - Forty years ago, the value of autopsies was widely recognized as new diseases were discovered or clarified and scientific technology advanced greatly. Despite the autopsy's strong foundation, its value is not currently being properly conveyed to physicians or patients. Although autopsy-related policy exists, these policies have had little effect on increasing or even maintaining adequate autopsy rates. More recently, the autopsy has fallen on hard times, with US hospital rates now below 5%. The reasons for the decline in rates are multifaceted and include a lack of direct reimbursement for the procedure, lack of defined minimum rate standards, overconfidence in diagnostic technology, and the fear of litigation. Regardless of the reasons for the declining rates, the ethical and professional reasons for increasing the number of autopsies are far more important. PMID- 11941996 TI - Left ventricular ejection fraction test rates for Medicare beneficiaries with heart failure. AB - The left ventricular ejection fraction (LVEF) test rate is increasingly used as a quality of care indicator for patients with heart failure. Our study produced benchmark LVEF test rates in a Medicare fee-for-service population for consideration by a clinical panel assembled by the Health Care Financing Administration. Our sample consisted of 46,583 beneficiaries admitted to the hospital for heart failure and with a complete set of Medicare fee-for-service bills dated 1996 or 1997. The national 2-year LVEF test rate was 79% for Medicare fee-for-service beneficiaries hospitalized for heart failure. Except for 1 state, the test rate ranged from 61% to 89% across states. Our analysis demonstrates the feasibility of using billing data to compute LVEF test rates. Using a 2-year time window and measuring tests performed in outpatient as well as inpatient settings, we find a higher LVEF test rate than has been reported by most previous studies. PMID- 11941997 TI - A method for rapid acquisition of data from dispersed individuals. AB - We have developed a data-collection system that we believe has wide applicability in dispersed organizations that have different local information systems infrastructures. A questionnaire was developed using commonly available software tools and was implemented several times across a dispersed network of individuals to rapidly collect, organize, and analyze information with a minimum of secretarial time and administrative cost. A common e-mail database was created identifying all of the medical directors and care management directors in the Blue Cross Blue Shield system. Initial polling of these individuals yielded specific questions of interest, and a final questionnaire was developed. The focus of the project centered on the evaluation of disease state management initiatives within each of the plans and on the ongoing use and future potential for various medical management initiatives. A questionnaire was developed using a Microsoft Excel spreadsheet with the ultimate development of a database in mind. All of the questions, whether single or multiple answers, were mapped from question response sections to a hidden specified range created to import the flat file answer block into a database. Individual cells containing answers to questions were each mapped to a hidden area of the spreadsheet arrayed as a series of rows. As each questionnaire was returned to the central site, data was imported from the hidden range name into a Microsoft Access database. The process of collecting extensive information on a questionnaire-by-questionnaire basis took approximately 20 seconds of time per questionnaire. A final report was ultimately created composed of some 24 pages of detailed information on managed care across the participants, representing some 90% of the member organizations. Secretarial costs were needed only for final transcription of the report. PMID- 11941998 TI - [Prognostic value of peritoneal lavage cytology in gastric cancer]. AB - The microscopic detection of free peritoneal tumour cells in peritoneal lavage fluid in gastric cancer patients is a useful predictor of peritoneal recurrence and poor prognosis. The aim of this study was to verify the prognostic significance of intraoperative peritoneal lavage cytology and its value as a predictor of peritoneal recurrence. We evaluated the presence of free peritoneal tumour cells with extemporary cytological examination in a series of 170 peritoneal washing samples from patients undergoing gastrectomy for gastric cancer over the period from January 1992 to June 2001. Twenty-eight patients (16%) had positive extemporary lavage cytology and there were no false-negatives as compared with the final examination. All patients with positive cytology presented serosal infiltration (T3/T4). Positive peritoneal lavage cytology was a predictor of poor prognosis and peritoneal recurrence: the 24 month survival rate was 17% for positive and 60% for negative cases (P = 0.003); in positive cases 71% of recurrences were located in the peritoneum. Intraoperative cytological examination of peritoneal washings can detect the presence of free malignant cells in the peritoneal cavity and can be used to select patients who may benefit from intraperitoneal chemotherapy. PMID- 11941999 TI - [Acute torsion of the greater omentum: a case report]. AB - Taking as their starting point a case of greater omental torsion recently observed in their surgical department, the authors carefully review the relevant literature data and the aetiological, pathogenetic, clinical and diagnostic aspects which make this rare disease hard to identify preoperatively. They stress that resorting to laparotomy is a rational course of action even if laparoscopy today constitutes an effective diagnostic and therapeutic option. PMID- 11942000 TI - [Appendiceal mucocele]. AB - Mucocele of the appendix is a rare lesion, characterised by distension of the lumen due to accumulation of mucoid substance. Mucocele is often asymptomatic and is found incidentally during surgical explorations or ultrasonography studies. This report illustrates the clinicopathologic features of a case of appendiceal mucocele observed during an emergency laparotomy. The surgical treatment consisted in appendicectomy. Colonscopic examination subsequently revealed a left colonic precancerous lesion that was treated with endoscopic polypectomy. The case described underlines the importance of colonic surveillance in patients with mucocele. PMID- 11942001 TI - [Robotic techniques in laparoscopic surgery]. AB - Robotic technology is being increasingly used in surgery. The authors assess the usefulness, effectiveness and safety of the AESOP 2000 robotic device (Automated Endoscopic System for Optimal Positioning) in videolaparoscopic surgery. Two laparoscopic cholecystectomies were performed with the aid of the AESOP 2000 robot. A short increase in operative time was observed and there were no complications. Voice understanding was accurate and flawless and no inadvertent smearing of the lens occurred. The use of the AESOP 2000 robotic device is safe, improves the quality of vision and reduces the number of surgeons needed in the team. Nevertheless, a learning curve and simultaneous training in the use of the device are required. PMID- 11942002 TI - Simultaneous aortoiliac reconstruction and renal transplantation: is it safe? AB - A 54-year-old man with an aortoiliac aneurysm and renal failure due to renal artery thrombosis was placed on a transplantation waiting list. The aneurysm had a 3 cm diameter and, therefore, did not require aortoiliac reconstruction, while its evolution was followed by ultrasound color-doppler every six months. The aneurysm was stable and two years later, when a cadaver kidney became available, a preoperative ultrasound color-doppler showed initial wall dissection. Therefore, an abdominal aneurysmectomy using a standard Dacron bifurcation graft and renal transplantation were successfully carried out. The patient had no associated complications and 24 months after transplantation and aneurysmectomy currently has good renal function and distal pulses. Only 20 cases of simultaneous aortoiliac reconstruction and renal transplantation have been reported in the literature. The excellent results of our case and those reported in the literature prove that patients who have both severe aortoiliac disease and end-stage renal failure can safely undergo simultaneous aortic reconstruction and renal transplantation. However, the atherosclerosis in these patients is a generalized process, so that in the pretransplant protocol special attention should be paid to detecting coronary artery atherosclerosis. In fact, coronary artery disease may have a priority claim to therapy because of the high risk of myocardial infarction. Our own policy is to put the patient back on the waiting list for renal transplantation after treatment for coronary artery disease. Furthermore, considering that the management for aortoiliac disease and kidney failure is safe in both simultaneous and staged cases, we think that the real issue is whether or not these patients with coronary atherosclerosis can be candidates for renal transplantation. We believe that each transplant centre has to develop its own general policy for these critically ill patients on the basis of its own experience. PMID- 11942003 TI - [Cervico-mediastinal goiter]. AB - We analyzed 824 patients treated by thyroidectomy in our Institute from January 1995 to December 2000. We observed the presence of cervicomediastinal goiter in 117 patients (14.9%), 92.7% of whom presented euthyroidism. Sternotomies were performed in 15 patients (10 midline and 5 hemi-clamshell). On the basis of our own experience and the reports in the literature, we analyse the diagnostic and surgical approach to the therapy of cervicomediastinal goiter. PMID- 11942004 TI - [Lung resection: predictive value of respiratory function]. AB - The most important preoperative cardiopulmonary variables for identifying patients at increased risk prior to lung resection are: FEV1, FEV1-ppo, DLCO, MVO2, and SO2. The aim of this study was to evaluate the ability and usefulness of predictive postoperative FEV1 (FEV1-ppo) in 80 patients undergoing lung resection. Spirometry was performed before and 6 months after operation; residual respiratory function was calculated using Nakahara's formula, and data analysis calculations were performed using the chi 2 test. It was observed that the resulting predictive values were almost comparable to the values observed 6 months postoperatively in 63.75% of patients and the correlation proved statistically significant (P < 0.005). In view of its simple and rapid execution, we conclude that Nakahara's formula, compared with the others, remains a reliable standard method of assessing high-risk patients and planning appropriate surgery. PMID- 11942006 TI - [Surgical treatment of extracranial carotid aneurysms]. AB - The incidence of aneurysm ranges from 0.1 to 3.7 of all carotid artery diseases requiring surgery. The aim of the study was to describe our personal experience with the surgical therapy of aneurysms of the carotid artery. Over the period from 1980 to 2000 we observed 19 aneurysms of the extracranial carotid tract in 17 patients. Eleven patients were subjected to surgical treatment (M/F: 2:1; mean age: 59 years). The sizes of the lesion ranged from 2 to 5.5 cm. The locations of the lesions were: common carotid 5; internal carotid 4; bifurcation 4. A shunt was utilised in one patient. No mortality was recorded. In 2 patients there was a transitory central neurological deficit, and in 4 patients a transitory peripheral neurological deficit. A definitive peripheral deficit was recorded in one patient (9%). The authors recommend prompt surgical treatment, in view of the low morbidity and almost total absence of mortality. PMID- 11942005 TI - [Medullary carcinoma of the thyroid: a clinical contribution]. AB - The authors report 20 cases of medullary thyroid carcinoma over the period from 1980 to 2000. Two patients turned out to be inoperable and 18 patients underwent total thyroidectomy, associated with dissection of the central lymphatic compartment in 5 patients and with dissection of the central and lateral lymphatic compartments in 10 patients with clinical or instrumental evidence of cervical lymphadenomegaly. Serum calcitonin levels proved to be a reliable marker for the diagnosis of persistence or recurrence of the disease. The follow-up, lasting from 1 to 208 months, demonstrated that in 7 cases in which serum levels of calcitonin underwent normalization there was no recurrence of disease. Among 11 cases with persistence of high calcitonin levels, 6 died and only 2 presented no evidence of metastases. On the basis of our analysis of the cases reported, total thyroidectomy associated with dissection of the central lymphatic compartment is an adequate treatment for patients in stages I and II. The authors regard routine dissection of the lateral lymphatic compartment as unadvisable. PMID- 11942007 TI - [Gastric polyps: a clinical contribution]. AB - The incidence of diagnosis of gastric polyps is now higher than in past years owing to the introduction of endoscopy in the diagnosis and treatment of upper digestive tract disease. One hundred and sixty-four polyps removed from January 1984 to August 2000 were analyzed. The median age of the patients was 61.4 years (range: 16-84 yrs). Polypoid lesions were more frequent in males (M:F = 1.5:1). Seventy-nine patients were asymptomatic (48.2%). Sixty-four percent of the polyps were located in the antrum. We diagnosed 73 hyperplastic polyps, 27 adenomatous lesions, 8 inflammatory polyps and 56 pseudopolyps. Malignant lesions were detected in 9 adenomatous polyps (4 type I and 5 type II early gastric cancers). Endoscopy is the examination of choice in the diagnosis and treatment of gastric polyps. We confirm that there is a relationship between histological type, neoplastic change and the size of the polyps. PMID- 11942008 TI - [Severity of anemia and operative morbidity and mortality. Report on 3 clinical cases in Jehovah's Witnesses and review of the literature]. AB - It has been reported that patients who refuse blood transfusions, such as Jehovah's witnesses, can undergo major surgery. In a review of the literature, however, we critically examined the severity of anaemia in relation to operative mortality and morbidity rates. We report three cases of Jehovah's witnesses who underwent major surgery and presented complication during the postoperative period. Case 1: a 50-year-old man with oesophageal achalasia who underwent Heller's myotomy and Nissen's fundoplication. The postoperative period was complicated by massive haemorrhage and the patient was reoperated on postoperative day 1. After four years, he underwent total oesophagectomy because of severe chronic oesophagitis. On postoperative day 13 the patient suffered anteroseptal myocardial ischaemia, which was treated with medical therapy. Case 2: a 40-year-old man, admitted for ulcerative rectocolitis, who underwent total colectomy. On postoperative day 1 he presented massive haemorrhage and shock. He was reoperated and the postoperative period was complicated by myocardial ischaemia, renal failure and an enterocutaneous fistula. Case 3: a 65-year-old woman with ulcerative rectocolitis who underwent total colectomy and a temporary ileostomy. She suffered venous thrombosis of the lower limbs and pulmonary oedema. The patient died 14 months after surgery as a result of massive haemolysis by cryoagglutinins and cardiac arrest. PMID- 11942009 TI - [Laparoscopic approach in the treatment of perforated gastroduodenal ulcer]. AB - The authors report on their experience with the treatment of 34 consecutive patients suffering from perforated peptic ulcer with peritonitis. All patients has been treated by the laparoscopic approach using sutures according to the Graham-Steele technique and omental plication. Irrigation, suction of the entire abdominal cavity with isotonic sodium chloride solution and drainage completed the procedure. In three cases, after an initial laparoscopic operation, we converted to laparotomy because of a large or unusually positioned ulcer. The morbidity rate was low and one patient died of myocardial infarction. The mean hospital stay was seven days. The method, which requires good laparoscopic experience, is simple, allows thorough toilette, has a low morbidity and is safe for the patient. PMID- 11942010 TI - [3-trocar video laparoscopic cholecystectomy: a retrospective study]. AB - Laparoscopic cholecystectomy is the gold standard for lithiasis of the gallbladder. Over the period from 1991 to 2000, the authors performed 1,321 videolaparoscopic cholecystectomies in the Surgical Division of San Giovanni Battista Hospital in Zagarolo (Rome). From January 1998 to December 2000, 390 of these patients were operated on consecutively utilising only three trocars. The authors report on this restricted subgroup of patients and stress that videolaparoscopic cholecystectomy is the treatment of choice. They claim that routine perioperative cholangiography is useless and that a fourth trocar is unnecessary. PMID- 11942011 TI - [Intraoperative ultrasonography in the staging of pancreatic head neoplasms]. AB - Tumours of the head of the pancreas constitute the fourth most common cause of cancer deaths. These tumours are characterised by low survival rates (5% at 5 years) and low surgical resectability rates (20-25%). Liver metastases, lymph node and vascular involvement, and peritoneal metastases are, in our opinion, exclusion criteria for curative surgical resection. The aim of the study was to evaluate the impact of intraoperative ultrasonography on the staging of such tumours. Over the period from 1990 to 2000 we introduced intraoperative ultrasonography in the staging of pancreatic cancer. We evaluated 51 patients who at preoperative staging had been regarded as candidates for surgical therapy consisting in a pancreaticoduodenectomy. All patients had been staged by preoperative abdominal ultrasound, ERCP, CT and MRI. Intraoperative ultrasound and colour-Doppler imaging (from 1997 on) revealed involvement of (i) the liver, (ii) the splenomesenteric vessels and (iii) the portal vein. Intraoperative ultrasonography yielded a diagnosis of occult liver metastases in 10 cases and signs of vascular involvement (absence of cleavage, partial and total thrombosis) in 12. One false-negative was registered. Intraoperative ultrasonography in our experience showed 98% sensitivity and specificity in the detection of vascular and lymph-node involvement. Its sensitivity in the detection of liver metastases was 100%. Intraoperative ultrasound is a procedure with a very high sensitivity in the operative staging of cancer of the head of the pancreas. PMID- 11942012 TI - [A prospective study of 106 patients with hemorrhoids treated with PPH stapler. Early and long-term results]. AB - Muco-mucosal resection with a PPH stapler according to Longo allows repair of arterial hyperaemia, venous dilation and mucosal prolapse. From September 1 1997 to October 31 2000 a prospective study was conducted in 106 patients. The results show that the Longo haemorrhoidectomy technique is well tolerated and almost painless, and is characterised by a low complication rate and good results in the long term. PMID- 11942013 TI - Non-recurrent inferior laryngeal nerve. AB - Surgical damage to the inferior laryngeal nerve is one of the most feared complications of thyroid surgery. Prevention of surgical injuries requires systematic, early exposure of the inferior laryngeal nerve. A detailed knowledge of anatomical variations is necessary. Non-recurrent inferior laryngeal nerve is a rare anomaly on the right side and is exceptional on the left. Whereas the typical course of the inferior laryngeal nerve is due to the embryological development of the aortic arch and supra-aortic vessels, non-recurrence is associated with a vascular anomaly such as a right retro-oesophageal subclavian artery. The nervous anomaly on the left side is possible only with the occurrence of cardiac dextroposition (situs viscerum inversus) and a left retro-oesophageal subclavian artery. The situation is more dangerous when a non-recurrent branch of the inferior laryngeal nerve is associated with a recurrent branch. This anomaly does not appear to be associated with a vascular anomaly in all cases. The authors describe three cases of right non-recurrent inferior laryngeal nerve observed and one recent case of combined non-recurrent and recurrent nerve, highlighting the anatomical and surgical features of this anomaly. PMID- 11942014 TI - [Role of ultrasonographic imaging in the surgical management of acute diverticulitis of the colon]. AB - Diverticulosis of the colon is a very frequent pathology in the western word and is characterised by a high percentage of dangerous complications (10-25%). The most accurate method of staging diverticular disease is by CT scan. The aim of our study was to evaluate the sensitivity of ultrasonography in the evaluation and management of diverticular disease of the colon. We studied 51 patients: the parameters used to assess complicated diverticulosis of the colon were: 1) wall thickness; 2) presence of fluid collections and pericolic abscesses; 3) free liquid collections in the peritoneal cavity; 4) subdiaphragmatic free air; 5) presence of fistula tracts. Ultrasonography showed 66% sensitivity in the assessment of wall thickness and in detecting the presence of diverticula. The sensitivity rate was 100% in the detection of abscess complications, pericolic collections, free air and fistula tracts. False-negatives (5 patients) were all recorded in Hinchey stage I. The overall sensitivity amounted to 91%. In our experience the method is a first level examination in the approach to patients suffering from diverticular disease of the colon and presents high sensitivity and diagnostic accuracy. The method, in expert hands, is suggested as a first step in the clinical-diagnostic approach to patients suffering from acute abdomen due to diverticulitis of the colon. PMID- 11942015 TI - [Ambulatory saphenectomy: 80 operated cases using tumescent anesthesia]. AB - The authors describe their experience with local tumescent anaesthesia with lidocaine and bupivacaine in 80 cases of saphenous stripping. The method is discussed thoroughly and provides: 1) good anaesthesia throughout surgery, requiring only brief propofol sedation in 55 patients with large-volume legs; 2) less bleeding during and after surgery in all patients, with visual evaluation of ecchymoses at 3 and 7 days; 3) good postsurgical analgesia without functional impairment in all patients. These are real advantages in the ambulatory and day hospital surgery settings, inasmuch as the scheduled discharge time is not modified. Local tumescent anaesthesia proves to be a simple, easy technique, with good patient compliance; it is appreciated by the surgeon on account of its technical advantages and is helpful in the organization of modern surgical departments. PMID- 11942016 TI - [Radiofrequency ablation in primary and secondary liver tumors]. AB - The authors report the preliminary results of a new treatment for primary and secondary hepatic malignancies called radiofrequency ablation. Seven patients undergoing the new radiofrequency ablation procedure (4 M and 3 F; mean age 62 years) were studied. The 3 female patients were suffering from breast cancer metastases (1 case), left colon cancer metastases to the first hepatic segment with caval and portal compression (1 case), and metastases to the eighth hepatic segment from an operated left colon cancer (1 case). The 4 male patients were suffering from carcinoma of the first hepatic segment on a cirrhotic liver (1 case), two liver metastases from left a colon cancer (1 case), a single 3-cm diameter metastasis treated by laparotomic radiofrequency ablation after resection of a gastric cancer (1 case), and one metastasis to the eighth hepatic segment from an operated left colon cancer (1 case). Prior to treatment all patients were subjected to investigation of routine blood-chemistry parameters, hepatic enzymes, cancer markers (Ca 19.9, alphafetoprotein, cytokines), abdominal spiral CT or MRI or PET (18 FDG); and thorough ultrasonography. Four patients underwent percutaneous radiofrequency ablation; one patient with concomitant gastric cancer underwent laparotomic radiofrequency ablation and simultaneous removal of the tumour; one patient was treated by celioscopic radiofrequency ablation in the course of laparoscopic cholecystectomy; and one underwent transpleural-diaphragmatic radiofrequency ablation for metastases to the seventh and eighth hepatic segments. None of the patients undergoing ultrasonography and CT follow-up examinations after 6 months presented recurrence of hepatic metastases. This treatment, though its use has so far been limited to only a few cases with a short follow-up, opens up new prospects for the surgical treatment of primary and secondary malignancies, also in the light of experience based on a substantial number of patients, recently reported in the literature. PMID- 11942017 TI - Sclerosing encapsulating peritonitis: a rare complication of peritoneal dialysis. A case report. AB - Sclerosing encapsulating peritonitis is a typical, but at the same time, not so frequently observed complication of peritoneal dialysis. The aim of this article is to report on the authors' clinical and surgical experience with this disease. After a review of the international literature and a description of the typical clinical features of this disease, the authors describe a case of sclerosing encapsulated peritonitis observed in their surgical department, on its mode of onset (intestinal occlusion), clinical behaviour and surgical treatment. The treatment of the patient consisted in the removal of fibrous tissue, resection of a necrotic ileal loop and intestinal reconstruction by an end-to-end ileo-ileal anastomosis. From the technical point of view the result was good and resolution of the intestinal occlusion was obtained. Cardiocirculatory complications arose on 6th postoperative day were the cause of the patient's death. Sclerosing encapsulating peritonitis is a rare complication of peritoneal dialysis, but it should be borne in mind whenever a patient with a history of peritoneal dialysis reports episodes of abdominal pain, nausea and vomiting associated with weight loss. This kind of peritonitis may require surgical and non-surgical treatment, though laparotomy provides us with a reliable diagnosis and may be considered essential to resolve the intestinal obstruction. PMID- 11942019 TI - A case of giant retroperitoneal liposarcoma. AB - A 44-year-old female patient presented to us with fever and increased abdominal girth. A CT scan showed a very large retroperitoneal mass displacing the right kidney. At operation three enormous retroperitoneal tumours displacing the ascending colon and the right kidney and compressing the inferior vena cava were removed. Histopathological examination revealed a myxoid retroperitoneal liposarcoma. PMID- 11942018 TI - Autoimmune-like pancreatitis in thymoma with myasthenia gravis. AB - A patient with thymoma and myasthenia gravis admitted for surgery presented increased serum levels of pancreatic amylase and lipase. Suspecting a thymoma related autoimmune disorder, autoantibody serum titers were determined: increased autoantibody titers to acetylcholine receptors, thyroglobulin, thyroperoxidase and pancreatic insulin were detected. After thymectomy the serum levels of pancreatic enzymes decreased rapidly. Myasthenia gravis symptoms also improved. To the best of our knowledge no similar cases have been reported in the literature. PMID- 11942020 TI - [Mucin-secreting tumor of the appendix with large gelatinous effusion]. AB - The authors report a case of a mucinous tumour of the appendix with a large muco gelatinous effusion detected as a result of massive abdominal compression. The mucinous mass occupied the entire abdominal cavity and was compressing the abdominal vascular trunks. Increases in CEA, Ca 19.9 and Ca 50 were recorded. The abdominal CT scan was of fundamental importance for diagnosing the condition. Surgery was performed consisting in removal of the mucinous abdominal mass, the appendix and part of the caecum. The postoperative course was favourable with rapid restoration of abdominal organic functions and normalisation of CEA. The rarity of these tumours is stressed. PMID- 11942022 TI - An African therapy in dialogue with Freudian psychoanalysis. AB - This article summarizes and compares selected characteristics of Freudian psychoanalysis and of African psychotherapy. The author concludes that the Freudian emphasis on the individual and the African therapy's emphasis on the community may both lead to the positive restoration of both therapies' clients. PMID- 11942023 TI - Professional satisfaction among U.S. healthcare chaplains. AB - This article reports the results of a questionnaire sent to 1,099 chaplains with board certification from the Association of Professional Chaplains regarding determinants of professional satisfaction among U.S. healthcare chaplains. PMID- 11942024 TI - Caring for the stories that come to us: work narratives and their sacred promise. AB - People often lack opportunity and place to reflect upon and share the sacred moments that occur in their work. Thus, these experiences are marginalized and neglected. Yet, they embody the soul of one's vocation. Drawing upon material from narrative family therapy, this article suggests that attending to and remembering these core but marginalized experiences is an activity for pastoral care. Narratives integrate spirituality and work. The article discusses the structure of narrative, its life cycle, and living in a universe composed of many stories. It explains a storytelling ritual that creates a safe place among the listeners who can then receive the narrative's sacredness. Two extended examples of sacred work narratives are given. A task for pastoral care is to construct hospitable places, sanctuaries, to hear the voice of these marginalized stories. PMID- 11942025 TI - Pastoral counseling as an echoing process: a transformative therapy hour. AB - The author posits that the essential core of pastoral counseling is an echoing process between client and therapist and between diagnostic listening and restorative responding. He illustrates this process by drawing on the case of a depressed middle-aged client. The therapist's response is described as conveyed through personal presence that is welcoming, participative, courageous, and responsive. PMID- 11942026 TI - Mighty prophet/wounded healer. AB - The concept of wounded healer is revisited from the perspective of a pastoral counselor working in a pastoral counseling center, and this experience is generalized to pastoral caregivers working in other settings. Three types of wounds are reviewed: wounds resulting from the life experience of the caregiver, wounds resulting from listening to and containing the horrendous content and emotionally laden nature of client stories, and wounds brought about by doing our work within an unsupportive culture. Properly dealt with, these wounds offer a foundation of shared life experience connecting us with our clients. Coupled with messages clients bring us about hurtful and destructive aspects of our culture, these wounds also call us to become mighty prophets. In this role, we are urged to move outside our normal practice contexts into the larger community, and to speak out against social forces that hurt and destroy the selves and souls of persons. PMID- 11942027 TI - From object to subject: pastoral supervision as an intersubjective activity. AB - The author sketches the development of psychoanalysis in terms of its trends from notions of objectivity to intersubjectivity and how this movement can inform contemporary pastoral supervision. The article argues that these same trends may be noted in the development of contemporary educational and theological theories. PMID- 11942028 TI - A ceremony for grieving. PMID- 11942029 TI - Being present in the moment. PMID- 11942030 TI - Reflections on Easter. PMID- 11942031 TI - A question of numbers. PMID- 11942032 TI - The storyteller. PMID- 11942033 TI - Oxygenation measurements in head and neck cancers during hyperbaric oxygenation. AB - BACKGROUND: Tumor hypoxia has proven prognostic impact in head and neck cancers and is associated with poor response to radiotherapy. Hyperbaric oxygenation (HBO) offers an approach to overcome hypoxia. We have performed pO2 measurements in selected patients with head and neck cancers under HBO to determine in how far changes in the oxygenation occur and whether a possible improvement of oxygenation parameters is maintained after HBO. PATIENTS AND METHODS: Seven patients (five male, two female, age 51-63 years) with squamous cell cancers of the head and neck were investigated (six primaries, one local recurrence). The median pO2 prior to HBO was determined with the Eppendorf histograph. Sites of measurement were enlarged cervical lymph nodes (n = 5), the primary tumor (n = 1) and local recurrence (n = 1). Patients then underwent HBO (100% O2 at 240 kPa for 30 minutes) and the continuous changes in the oxygenation during HBO were determined with a Licox probe. Patients had HBO for 30 minutes (n = 6) to 40 minutes (n = 1). HBO was continued because the pO2 had not reached a steady state after 30 minutes. After decompression, patients ventilated pure oxygen under normobaric conditions and the course of the pO2 was further measured over about 15 minutes. RESULTS: Prior to HBO, the median tumor pO2 in the Eppendorf histography was 8.6 +/- 5.4 mm Hg (range 3-19 mm Hg) and the pO2 measured with the Licox probe was 17.3 +/- 25.5 mm Hg (range 0-73 mm Hg). The pO2 increased significantly during HBO to 550 +/- 333 mm Hg (range 85-984 mm Hg, p = 0.018). All patients showed a marked increase irrespective of the oxygenation prior to HBO. The maximum pO2 in the tumor was reached after 10-33 minutes (mean 17 minutes). After leaving the hyperbaric chamber, the pO2 was 282 +/- 196 mm Hg. All patients maintained an elevated pO2 for further 5-25 minutes (138 +/- 128 mm Hg, range 42-334 mm Hg, p = 0.028 vs the pO2 prior to HBO). CONCLUSIONS: Hyperbaric oxygenation resulted in a significant increase in the tumor oxygenation in all seven investigated patients. A significant increase at the point of measurement could be maintained for several minutes after decompression and after leaving the hyperbaric chamber. PMID- 11942034 TI - [Effectiveness of postoperative radio- and chemotherapy in N(+) esophageal carcinoma]. PMID- 11942035 TI - [Subclinical heart damage caused by adjuvant anthracycline chemotherapy]. PMID- 11942036 TI - [Superiority of letrozole compared with tamoxifen as first line therapy of postmenopausal women with advanced breast cancer: results of a phase III study of the International Letrozole Breast Cancer Group]. PMID- 11942037 TI - [Value of centralization of patient management exemplified by breast carcinoma]. PMID- 11942038 TI - [Recommendations for treatment with permanent, interstitial brachytherapy alone in locally limited prostate cancer]. PMID- 11942039 TI - [Combined radiotherapy and gemcitabine. Evaluation of clinical data based on experimental knowledge]. AB - BACKGROUND: In experimental studies the nucleoside analog Gemcitabine (2',2' difluorodesoxycytidine) clearly demonstrates radiation enhancing properties. After describing the pharmacological Gemcitabine-related data and the clinical studies regarding combined radiochemotherapy and taking under consideration the in-vitro data and the results provided by animal models, this overview is aimed to draw clinically relevant conclusions, resulting in the improvement of treatment approaches. MATERIALS AND METHODS: The available literature data regarding the metabolism and the mechanism of action, the evaluation of possible schedules of administration, and combined radiochemotherapy including Gemcitabine has been reviewed. Publications reporting experimental data in vitro and in vivo as well as our own experimental results have been incorporated. RESULTS: In clinical phase I and II studies, the favorable tumor response is accompanied by a high incidence of grade III-IV toxicities whereby the maximum-tolerated dose (MTD) of the various schedules of administration used is always lower compared to the MTD of single-agent treatment. In in-vitro and in-vivo data addressing the description and the evaluation of the radiation enhancing mechanism (especially influence on cell cycle, depletion of the dATP pool, induction of apoptosis, inhibition of DNA synthesis, reduction of DNA repair) this effect is already observed with non and moderately toxic Gemcitabine concentrations and depends on drug concentration and exposure time. Independent of the fractionation effect of radiotherapy, the radiation enhancement is persistent for at most 72 hours after the end of drug exposure. Taking under consideration the single dose per day and the target volume, a prolonged infusion and/or a twice-weekly administration of Gemcitabine at low concentration each and simultaneous radiotherapy are presumably considered to resemble the experimental data. CONCLUSION: It is without doubt that data provided by clinical studies are of highest relevance for the evaluation of an optimized radiochemotherapy with Gemcitabine. However, although it is often difficult to transfer experimental data into the clinical situation, these data offer the possibility to develop an improved schedule of administration in patient treatment based on rational evidence in tumor biology. PMID- 11942040 TI - [Adjuvant simultaneous radiochemotherapy after operated uterine cervix carcinoma in high risk situation. Results of a pilot study]. AB - BACKGROUND: The most important factors for prognosis of cervical cancers are age and histological criteria such as the tumor size, the involvement of lymph nodes, lympho-vascular space involvement as well as microvessel involvement and poor tumor differentiation (grading 3). Here we present the results of concomitant chemo-radiation at high-risk situation of patients with cervical cancer after surgery. PATIENTS AND METHODS: The study comprised 34 patients with median age of 40 years (26-63 years) after Wertheim surgical technique for cervical cancer at the FIGO Stages IB (n = 19) and IIB (n = 15). All patients were treated between November 1995 and June 1999 by a schedule of concomitant chemoradiation. The indication for this treatment was given by the positive histological proof of lymph node metastasis, microvessel or lympho-vascular space involvement as well as a G3 grading. The chemo-therapy was given in week 1 and 5 (day 1-5 and day 29 33). The dosage of cisplatin was 20 mg/m2/d on every day and 5-FU was given as a 120-h infusion with 600 mg/m2/d. The external beam radiotherapy was applied to the pelvis with 1.8 Gy per fraction up to 50.4-54 Gy. In two patients the paraaortal region was irradiated too because of the involvement of these lymph nodes. RESULTS: The median observation time was 48 months (3-68 months). 30 patients are alive (88%) in complete response. Four patients died. The mean survival was 61 +/- 3 months. We have seen only slight acute toxicities of grade 1 and 2. Three patients suffered from a grade 3 diarrhea and three patients developed a grade 3 leukopenia. In seven patients we found a secondary lymphedema as a late toxicity. CONCLUSION: The concomitant chemoradiation containing cisplatin in high-risk situation for cervical cancer after surgery improves the outcome and survival in these patients. PMID- 11942041 TI - The role of radiation therapy in the management of desmoid tumors. AB - PURPOSE: To investigate the role of radiation therapy (RT) in the management of desmoid tumors. PATIENTS AND METHODS: Retrospective analysis was performed on 28 patients with desmoid tumors treated with radiation therapy between March 1989 and March 1999. Tumor site was intraabdominal in three, abdominal wall in three and extraabdominal in 22 patients. Median tumor dose was 48 Gy (range 36-60 Gy). Radiation therapy was delivered postoperatively in 26 of 28 patients, two patients received radiation therapy for unresectable recurrent tumors. RESULTS: Median follow-up was 46 months (range 13-108 months). Actuarial 5-year control rate was 73%. We observed six recurrences, located within the radiation field in one patient, out of field in two and at the field margin in three patients. All patients with intraabdominal tumors have been controlled without severe side effects. CONCLUSIONS: Radiation therapy is an effective treatment after incomplete resection of desmoid tumors. We did not observe a benefit for tumor doses exceeding 50 Gy. In some patients with circumscribed intraabdominal desmoid tumors, radiation therapy might be a treatment option with low toxicity, if 3-D treatment planning is utilized. PMID- 11942042 TI - [Positioning accuracy in conformational prostatic irradiation using portal imaging]. AB - BACKGROUND: Conformal radiotherapy techniques as used in prostate treatment allow to spare normal tissue by conforming the radiation fields to the shape of the planning target volume (PTV). To be able to fully utilize the advantages of these techniques correct patient positioning is an important prerequisite. This study employing an electronic portal imaging device (EPID) investigated the positioning uncertainties that occur in the pelvic region for different patient positioning devices. PATIENTS AND METHODS: 15 patients with prostate cancer were irradiated with or without rectal balloon/pelvic mask at a linear accelerator with multileaf collimator (MLC). For each patient multiple portal images were taken from different directions and compared to the digitally reconstructed radiographs (DRRs) of the treatment planning system and to simulation films (Table 1, Figure 1). RESULTS: In spite of different positioning devices, all patients showed comparable total positioning uncertainties of 4.0 mm (lateral), 4.5 mm (cranio caudal) and 1.7 mm (dorso-ventral). The lateral positioning error was reduced for the pelvic mask patients while the cranio-caudal error increased (Table 2, Figure 2). A systematic and a random component sum up to the total positioning error, and a good estimate of the magnitudes of the two is possible from six to eight portal images (Figure 3). CONCLUSIONS: With a small number of portal images it is possible to find out the systematic and random positioning error of a patient. Knowledge of the random error can be used to resize the treatment margin which is clinically relevant since this error differs greatly for different patients (Figure 4). Image analysis with EPID is convenient, yet has some problems. For example, one only gets indirect information on the movement of the ventral rectum wall (Figure 5). The successful operation of positioning devices, although, needs further improvement--especially if one focuses on IMRT. PMID- 11942043 TI - Influence of the leaf extract of Mentha arvensis Linn. (mint) on the survival of mice exposed to different doses of gamma radiation. AB - BACKGROUND: The aim of the present study was to evaluate the radioprotective effect of Mentha arvensis (mint) on the survival of mice exposed to various doses of whole-body gamma radiation. MATERIAL AND METHODS: The radioprotective effect of various doses (0, 2.5, 5, 10, 20, 40 and 80 mg/kg body weight) of chloroform extract of mint (Mentha arvensis Linn.) was studied in mice exposed to 10 Gy gamma radiation. RESULTS: The 10 mg/kg of mint extract was found to afford best protection as evidenced by the highest number of survivors in this group at 30 days post-irradiation, and further experiments were carried out using this dose of mint extract. The mice treated with 10 mg/kg body weight mint extract or oil were exposed to 6, 7, 8, 9 and 10 Gy of gamma radiation and observed for the induction of radiation-sickness and mortality up to 30 days post-irradiation. The mint extract pretreatment was found to reduce the severity of symptoms of radiation sickness and mortality at all exposure doses and a significant increase in the animal survival was observed when compared with the oil + irradiation group. All of the animals that were treated with 10 mg/kg mint extract and then exposed to 7 Gy irradiation were protected against the radiation-induced mortality when compared with the concurrent oil + irradiation group, in which 20% animals died by 30 days post-irradiation. The mint extract treatment protected the mice against the gastrointestinal death as well as bone marrow deaths. The DRF was found to be 1.2. The drug was non-toxic up to a dose of 1,000 mg/kg body weight, the highest drug dose that could be tested for acute toxicity. CONCLUSION: From our study it is clear that mint extract provides protection against the radiation-induced sickness and mortality and the optimum protective dose of 10 mg/kg is safe from the point of drug-induced toxicity. PMID- 11942044 TI - [Treatment of keratitis superficialis chronica of the dog with strontium 90]. AB - BACKGROUND: Corneal pannus is a disease which, if untreated, nearly always is progressive and may lead to blindness of the affected dog. A therapeutic standard is yet to be defined. Beta-ray irradiation with Sr-90 is often recommended on a casuistic basis, but systematic studies are sparse. The aim of the present study was to evaluate efficacy and to document side effects of radiotherapy with Sr-90. MATERIAL AND METHODS: 17 animals were treated. 13 of them received treatment of 15 Gy surface dose twice within 2 days with additional medical therapy with ciclosporin and prednisolon. Only the more affected eye was treated with radiation which was applied with an eye-applicator, the other eye served as control. Four animals with already advanced impairment of vision received keratectomy, afterwards radiation was applied on both sides. RESULTS: Medical treatment alone led to deterioration in vascularization and spread of pigmentation in eleven of 13 (85%) of the control-eyes, density of pigmentation increased in eight of 13 (62%). After radiation therapy, almost all animals showed a marked initial improvement. Even if progressive disease occurred later on, further worsening as it happened in the control-eyes could be stopped in nine resp. ten of 13 eyes (69% and 77%). All animals with keratectomy and radiotherapy regained and preserved adequate vision. Besides short-term blepharospasm, no side effects were recorded. CONCLUSION: Corneal pannus is responsive to radiation therapy with Sr-90 and long-term benefit can be achieved. Side effects are minimal. Optimal sequencing of therapy and dosage still have to be examined. PMID- 11942046 TI - Prevention of liver cancer: current strategies and future perspectives. PMID- 11942045 TI - Chemoprevention of colorectal carcinogenesis. AB - The field of cancer prevention is advancing rapidly, largely owing to post genomic technology that has revolutionized our ability to identify and characterize molecular profiles for cancer. Advances in colorectal cancer screening (e.g., endoscopy, fecal occult blood testing, and mutational analysis) have made the detection and eradication of preinvasive neoplastic lesions the standard of care. Basic and translational sciences are building on these advances, and continue to expose molecular hallmarks of carcinogenesis that can be exploited as targets for molecularly targeted preventive interventions (i.e., chemoprevention). These targets will help identify more effective and better tolerated preventive agents. Carcinogenesis is now recognized as a disease in itself and has become the target of an ever-expanding array of preventive interventions. PMID- 11942047 TI - The significance of telomerase activity in cancer lesions and the noncancerous epithelium of the esophagus. AB - BACKGROUND: It is well known that telomerase activity is expressed in cancer cells. A recent study has revealed that weak telomerase activity was also detected in normal cells. Telomerase activity is thus thought to be useful as a diagnostic biomarker. In the present study, we studied the difference in telomerase activity between cancer tissue and noncancerous epithelium tissue of the esophagus. METHODS: One hundred and thirty specimens of esophageal cancer tissue were collected at surgery and analyzed in a telomeric repeat amplification protocol (TRAP) assay. Telomerase was expressed quantitatively as the total product generated (TPG) value. RESULTS: Telomerase activity was detected in all esophageal cancer tissues and in 87% of the noncancerous epithelium specimens. The mean TPG values in the cancer tissue and the noncancerous epithelium specimens were 49.5 +/- 39.5 (mean +/- 1 SD) and 11.5 +/- 5.9, respectively, with a significant difference (P < 0.0001). CONCLUSION: The telomerase activity in esophageal cancers was significantly higher than that in the noncancerous epithelium. From the mean value plus 1 SD for noncancerous epithelium, the cutoff value was set at 17.4 as an esophageal cancer-specific diagnostic marker. This cutoff value showed a high predictive value, specificity, and sensitivity, of greater than 80% for each category. These results suggest that this cutoff value could be a useful biomarker for the detection of esophageal carcinoma. PMID- 11942048 TI - Establishment of an immortalized T-cell line from lung cancer tissue: phenotypic and functional analyses. AB - BACKGROUND: In order to investigate host defense against solid tumors, valuable information could be provided by ex-vivo analyses of functional immune cells in tumor tissues. However, available sources of fresh tumor-infiltrating T cells (TIL) are usually very limited, and it is often difficult to establish TIL lines. In this study, we analyzed the phenotypic and functional characteristics of TIL, using an immortalized cell line prepared by cotransfection of human c-myc and c Ha-ras. METHODS: A human T-cell line was established by cotransfecting c-Ha-ras and c-myc oncogenes to T lymphocytes freshly isolated from human lung large-cell carcinoma tissue. The phenotypes were assessed by flow cytometry. T-cell receptor (TCR) V alpha- and beta-usage was analyzed by polymerase chain reaction (PCR) and Southern blot. Cytotoxic activity against autologous and allogeneic tumor cell lines was examined by standard 51Cr-release cytotoxicity assays. Cytokine production by the established T-cell line, 904-T1, in response to stimulation by autologous tumor cells was assayed by using an enzyme-linked immunosorbent assay. RESULTS: 904-T1 (CD3+, CD8+, CD56+, CD16-, CD161-, TCR V alpha 9, 13, and V beta 1, 5) displayed a broad range of MHC-nonrestricted tumoricidal activity against various human tumor cell lines, but did not lyse autologous B cells transformed by Epstein-Barr virus. The cytotoxicity of 904-T1 was not mediated by a T-cell antigen receptor or by Fas-ligand, but by perforin-based cytolytic pathways, and was enhanced by interleukin (IL)-12. 904-T1 cells produced large amounts of interferon (IFN)-gamma, but not tumor necrosis factor (TNF)-alpha or IL-4 in response to autologous tumor cells, and produced high levels of IFN-gamma and TNF alpha, and a substantial level of IL-4 following stimulation with anti-CD3 monoclonal antibody. CONCLUSIONS: Our results suggest that 904-T1 cells were natural killer T (NKT)-like cells with regard to their non-specific killing, cytokine repertoire, and sensitivity to IL-12, although the repertoire of the TCR variable region was not compatible with that of NKT cells. PMID- 11942049 TI - Survival of patients with advanced ovarian cancer treated with intermittent chemotherapy following cytoreductive surgery and adjuvant chemotherapy. AB - BACKGROUND: The purpose of this study was to report the duration of the progression-free interval (PFI) in advanced ovarian cancer patients who were treated with intermittent maintenance chemotherapy. METHODS: Between 1991 and 1998, 25 patients with stage III or IV ovarian cancer were enrolled in a trial of intermittent maintenance chemotherapy. All patients underwent cytoreduction surgery, and received adjuvant chemotherapy, after which they were treated with intermittent maintenance chemotherapy every 3 to 4 months for 2 years. RESULTS: The median PFI in the 25 women in the intermittent chemotherapy group was 25 months, while in the 32 patients in the control group it was 18 months (P = 0.0124). The median survival of women treated with the intermittent chemotherapy was 34 months, and for the control group patients, it was 35 months (P = 0.0672). Multivariate analysis in the intermittent chemotherapy group revealed that the only factor that correlated significantly with PFI was the status after adjuvant chemotherapy (P = 0.0137). In patients with no evidence of disease after the adjuvant chemotherapy, the median survival was 39 months in the intermittent chemotherapy group, and 35 months in the control group (P = 0.0156). The median PFI was 28 months in the intermittent chemotherapy group, and 18 months in the control group (P = 0.0012). CONCLUSION: It would be warranted to perform intermittent maintenance chemotherapy for patients with advanced ovarian cancer, if a clinically disease-free status could be achieved after completion of the standard treatment procedure. PMID- 11942050 TI - "In situ preparation": new surgical procedure indicated for soft-tissue sarcoma of a lower limb in close proximity to major neurovascular structures. AB - BACKGROUND: When soft-tissue sarcomas occur near neurovascular structures, preoperative images cannot always reveal the accurate relationship between the tumor and these structures. Therefore, in some patients, neurovascular structures are sacrificed unnecessarily. In other patients, neurovascular structures are preserved with an inappropriate margin, followed by local recurrence. The objective of this study was to evaluate a new surgical method, "in situ preparation" (ISP), which enables the preparation of neurovascular bundles and the intraoperative evaluation of the surgical margin without contamination by tumor cells. With this method, additional procedures, including pasteurization, alcohol soaking, and distilled water soaking of the preserved neurovascular bundle can also be performed to preserve the continuity of vessels. METHODS: Between April 1992 and December 1998, 18 patients with soft-tissue sarcoma were operated on using ISP. The tumor and neurovascular structure were lifted en bloc from the surgical bed and separated from the field by the use of a vinyl sheet. The consistency of the neurovascular structures was preserved. The tissue block could be freely turned around and the neurovascular structure was separated from the block through the nearest approach. The margin between the tumor and neurovascular structure was evaluated, and an additional procedure, such as pasteurization, alcohol soaking or distilled water soaking, was performed, according to the safety of the surgical margin. RESULTS: Only one patient showed recurrence after ISP. Complications after ISP were arterial occlusion in two patients and nerve palsy in three patients. The main cause of these complications was the long period of pasteurization; modified additional procedures could prevent such complications. CONCLUSIONS: ISP is a useful method with which to ensure a safe surgical margin and good functional results. PMID- 11942051 TI - A long-term survivor of leiomyosarcoma around the right side of the base of the skull: effective radiotherapy combined with intra-arterial chemotherapy. AB - We report a rare case of a leiomyosarcoma that developed around the right side of the base of the skull in a 51-year-old woman. The patient consulted our hospital complaining of pain in the right side of her neck and upper right arm in August 1994. A leiomyosarcoma, originating around the right side of the neck and base of the skull was diagnosed. Initially, surgery was planned, but invasion into the spinal canal was discovered. Curative resection of the leiomyosarcoma around the right side of the base of the skull was not possible. Therefore, external beam radiotherapy (EBRT) combined with intra-arterial chemotherapy and hyperthermia was employed. After the treatment, the tumor decreased in size to 45% of its initial volume, and, simultaneously, her symptoms completely disappeared. The patient initially remained clinically free of the disease, but showed reaggravations at the primary tumor site 3 years and 3 months, and 4 years and 11 months, after the first treatment. The reaggravations were treated with EBRT combined with intra-arterial chemotherapy. As a result, she survived for 5 years and 7 months after the first treatment. PMID- 11942053 TI - [Sociology. The development of temporary lodging]. PMID- 11942052 TI - Aortitis during intraarterial chemotherapy for cervical cancer. AB - A 76-year-old woman with stage IIb cervical cancer with a bulky tumor experienced aortitis during continuous intraarterial cisplatin-based chemotherapy. The chemotherapy was administered through a catheter tip placed in the aorta abdominalis, utilizing an external infusion pump. During the third course of chemotherapy, she complained of left-sided lower back pain and moderate fever was observed. Elevated white blood cell count (WBC) and C-reactive protein (CRP) level were noted, and an abdominal X-ray and urgent computed tomography (CT) were performed. The catheter tip was displaced against the arterial blood flow. At this level of the aortic wall, soft tissue density surrounded the aorta completely. Aortitis caused by the intraarterial chemotherapy, was strongly suspected. It was thought that the maldistribution of drugs and changes in the drug flow occurred due to the vertebral height movement of the catheter tip against the aortic blood flow, and there, flow to the vasa vasorum may have occurred. Chemical vasculitis of the vasa vasorum due to the anticancer drugs was strongly suspected as a contributing factor of the aortitis. Because of the long term use of an intraarterial catheter, the maldistribution of drugs and changes in the drug flow occurred physically and biologically during the course of the chemotherapy. We recommend occasional monitoring of the location of the catheter tip and a repeat evaluation with contrast medium in regard to flow to the vasa vasorum. PMID- 11942054 TI - [Investigation. Temporary reception, a means of institutional integration?]. PMID- 11942055 TI - [Strategies of care. Temporary reception, instead of proximity]. PMID- 11942056 TI - [Strategies of care. The right to "breathe" of families and of the aged]. PMID- 11942057 TI - [The right to "breathe" of families and of the aged. An example of collaboration between the hospital and its partners in Val-d'Oise]. PMID- 11942058 TI - [The senses and animation]. PMID- 11942059 TI - [The main point. The arguments, benefits, risks and recommendations]. PMID- 11942060 TI - [Strategies of care. The problems of deglutition in the aged]. PMID- 11942061 TI - [Medical strategies. Longevity and old age, scientific and medical aspects]. PMID- 11942063 TI - [Osteoporosis, a current target of interest]. PMID- 11942062 TI - [Preventing elder abuse]. PMID- 11942064 TI - [Care of the aged]. PMID- 11942065 TI - [Nursing practice. Psychosocial emergencies and team work]. PMID- 11942066 TI - [Nursing care strategy. Role of the nurse in team work]. PMID- 11942067 TI - [Prevention. National education, a team approach in child and adolescent psychiatry]. PMID- 11942068 TI - [Nursing practice. Prevention in mental health in colleges and lyceum]. PMID- 11942070 TI - [Reflection. The nurse in psychiatry, evolution of employment or new profession?]. PMID- 11942069 TI - [Community service, beyond conceptual design]. PMID- 11942072 TI - [Nursing practice. Loss of contact with reality in schizophrenia]. PMID- 11942071 TI - [Nursing practice. Therapeutic involvement in a trial]. PMID- 11942073 TI - [Horn GKPS youth project: pets in geriatric nursing]. PMID- 11942074 TI - [Shift work in nursing care]. PMID- 11942075 TI - [When nursing care hurts.. It is part of daily experience to be confronted with one form ov violence or another. Student project in nursing research curriculum]. PMID- 11942076 TI - [Breast carcinoma. A topic that affects us women! Because cases of breast carcinoma occurred in our family circle, we became interested in information regarding risk factors, prevention, mass screening and counseling facilities. School project in nursing research curriculum]. PMID- 11942077 TI - [Conversion of nursing diagnoses as an integrated element of the nursing process]. PMID- 11942078 TI - [The genotype of the principal Mycobacterium bovis in Argentina is also that of the British Isles: did bovine tuberculosis come from Great Britain?]. AB - Bovine tuberculosis is a highly prevalent animal disease in Argentina. In this work evidence was obtained showing that a major Mycobacterium bovis group in Argentina had been introduced with the bovine bulls imported from the United Kingdom at the end of the XIX century. This evidence came from two sources: historical, obtained by bibliographical references, and from laboratory results, using a molecular typing method called spoligotyping. These strains are also present in other countries that introduced cattle from the same origin. PMID- 11942079 TI - [Production of pectinases by Penicillium simplicissimum A3263 in an amaranth-seed flour medium]. AB - The present work studies the production of pectinases using a strain of Penicillium simplicissimum A3263 and considering the influence of adding Amaranthus cruentus seed meal in a selected medium. We also considered the influence of aeration on enzyme production. Research was oriented towards the production of pectin lyase, the enzyme having the highest commercial value. This work was carried out in Erlenmeyer flasks in rotary shaker to select the medium and in a mechanically stirred fermentor to study aeration. The microorganism was developed as pellets of 1 mm diameter. Enzyme levels were of the order of 8216.21 pectin lyase units and 167.57 polygalacturonase units per gram of fungal biomass, respectively, using a medium containing 40 g/l of amaranth seed meal. As for the influence of aeration, it was determined that the higher values were obtained at 750 rpm corresponding to an oxygen absorption rate of 2691 ml O2/lh for an air flow of 1 l/l.min. The results obtained are considered very important in view of the fact that they exceeded in 550% those obtained by other authors. PMID- 11942080 TI - [Candida carriage in the oral mucosa of a student population: adhesiveness of the strains and predisposing factors]. AB - The aim of this study was to establish oral carriage of Candida and possible factors associated to their virulence in young adults and their relation with local and general situations considered as predisposing factors. Samples were obtained from dorsum tongue in 70 students attending the Faculty of Dentistry (University of Buenos Aires) average age: 23, all in healthy oral conditions. Of these, 21.42% were Candida positive. These samples were seeded in CHROMagar. Candida identification was completed in milk agar and Fungichrom 1. The following species were identified: 11 Candida albicans (C.a), 2 Candida parapsilosis (C.p) and 1 Candida glabrata (C.g). In one case, 2 species (C.a and C.g) were isolated in the same sample. Virulence was determined as adherence capacity by biofilm or in vitro plaque formation and hydrophobicity. Different host factors were analyzed statistically to establish their importance as predisposing factors to allow Candida colonization. Adherence of C.a. was found to be similar in all C.a. strains, whereas significant differences were found between C.a. and C.p. and between C.a. and C.g. Only the antiseptic mouthrinse and the diet were significant among the considered factors. PMID- 11942081 TI - [Electrophoretic karyotypes and genomic DNA restriction fragment analysis: their usefulness as tools in the epidemiological study of Candid parapsilosis]. AB - During the past decades, several studies have reported an increase in the incidence of nosocomial candidosis. In a prospective study, performed at the Departamento de Micologia, INEI, ANLIS Dr. C. G. Malbran and the Servicio de Neonatologia and Microbiologia, Hospital de Ninos Sor Maria Ludovica, from October 1995 to December 1996, 167 patients with candidosis were detected. Candida species isolated were C. albicans (53.1%), C. parapsilosis (26.5%) and C. tropicalis (14.8%). The aim of this work was to characterize the clinical C. parapsilosis isolates from pediatric patients hospitalized in two neonatal intensive care units from the same hospital and to evaluate the usefulness of electrophoretic karyotype (EK) and restriction endonuclease analysis of genomic DNA (REAG) using a low frequency digestion enzyme. EK of all isolates disclosed 12 banding patterns and REAG with endonuclease Sfi I showed only 5 groups. However, isolates from the control group could not be separated from the clinical isolates. The isolates within each dendogram group for EK or REAG were apparently unrelated. Our results show that EK yields better results than REAG, but that it falls short of the desired discrimination, which suggests that these techniques do not seem to be useful for studying nosocomial C. parapsilosis outbreaks. PMID- 11942082 TI - [Effect of cocultivating fungal species on the degradation of lignocellulose residues]. AB - The degradation potential of Phanerochaete sordida, Trametes trogii, Coprinus truncorum and Paecilomyces sp. upon yard wastes was evaluated. The species had been inoculated individually or in pairs formed by P. sordida and Paecilomyces sp., T. trogii and Paecilomyces sp., and C. truncorum and Paecilomyces sp. The highest level of endoxilanase activity was produced by P. sordida growing alone, during day 21 (1.09 U/g of dry material), but in P. sordida and Paecilomyces sp. cultures, the detected activity did not overcome 0.27 U/g of dry material during the whole experiment. T. trogii showed maximum activity on day 14 (0.78 U/g of dry material), but in T. trogii and Paecilomyces sp. cultures, the values increased until day 21 (1.07 U/g of dry material). P. sordida endocellulase activity reached its maximum on day 28 (0.08 U/g of dry material), but in P. sordida and Paecilomyces sp. cultures, this activity increased during the whole experiment (0.04 U/g of dry material). The major weight loss was found in P. sordida (27.6%). The possible beneficial effect of co-culture in yard wastes biodegradation is discussed. PMID- 11942083 TI - [Study of Mycobacterium bovis in milk using bacteriological methods and the polymerase chain reaction]. AB - The frequency of Mycobacterium bovis detection in milk samples obtained from infected animals was explored in an intensive dairy area in Argentina. To this end, an "in house" polymerase chain reaction (PCR) method was developed using Mycobacterium tuberculosis Complex specific INS1-INS2 primers, and its performance was compared with that of bacteriological methods. The decontamination procedures previous to culture reduced M. bovis viability. The pathogen was identified in milk samples from 1 of 143 infected cows and in none of 43 uninfected ones. Even though PCR sensitivity was found to be 2-20 times higher than that of bacteriology in experimentally inoculated milk samples, all 186 field samples resulted negative by PCR, including the bacteriologically confirmed one. In spite of the high prevalence of bovine tuberculosis in Argentinian dairy herds, the detection of M. bovis in milk is an unusual finding. PMID- 11942084 TI - [Comparison of different detection methods for methicillin resistance in Staphylococcus aureus]. AB - The investigation of methicillin resistance in Staphylococcus aureus (MRSA) is a serious problem for the physician and microbiologist. Accurate and rapid detection is essential for the use of appropriate antimicrobial therapy and for the control of nosocomial spread of the resistant strain. The performance characteristics of the MicroScan Overnight Conventional Pos Combo 12 panels (MOCP), BBL Crystal MRSA ID (CR), E-test and agar screen plate (Muller Hinton agar with oxacillin 6 micrograms/ml and 4% NaCl) (AS) were evaluated for the detection of oxacillin resistance. Thirty S. aureus clinically significant strains with different PFGE (Pulse Field Gel Electrophoresis) banding pattern were tested, and 22 of them were mecA positive by PCR. These strains were also analyzed by mecA and Tn554 polymorphism. All mecA positive strains were classified as methicillin resistant by MOCP and E-test. CR and AS failed to detect oxacillin resistance in 2 strains. One false positive was only detected by E-test. Accurate testing for the presence of MRSA may reduce the need for empiric therapy with vancomycin for patients with staphylococcal infections. According to our results the best performance was obtained with MOCP. However, as a rapid method, CR gave acceptable sensitivity for clinical purposes. PMID- 11942086 TI - Use and barriers to use of laboratory data by clinical dietitians. AB - The results of this study reveal two general areas of concern for clinical dietitians in the use of laboratory data--their lack of confidence in their own interpretive skills and the perceived resistance of physicians. Although clinical dietitians receive theoretical and applied training on the use of laboratory values in their undergraduate education and dietetic internship, clinical managers should be aware that they need support and continuing education to use their laboratory assessment skills to the fullest. Also, dietitians should demonstrate more clearly to physicians their knowledge, skill, and needs in this area. They should initiate and continue dialogue about the importance of laboratory analysis to nutrition care and improved patient outcome. Evidence based data and clear documentation of improved medical outcomes should help overcome barriers to use of laboratory data by clinical dietitians. PMID- 11942085 TI - Genomic variations among wildtype and mutant strains of pseudorabies virus. AB - Both wild-type virulent and mutant strains of pseudorabies virus (PrV) were used in this study. Mutants used were derived from the plaque purified strain PrVmAIP. A total of six drug resistant mutants, three bromodeoxyuridine (BUdR) resistant and three iododeoxyuridine (IUdR) resistant, respectively, were isolated and passaged in chicken embryo fibroblast (CEF) cells. The DNA of these PrVs were compared with the wild-type isolates by means of the restriction fragment pattern (RFP) findings produced with Bam HI, Kpn I, Hind III and Bgl II restriction enzymes (RE). Compared to the wild-type PrVs (PrV-VBA1-parental strain of PrVmAIP; PrV-VBA2; PrV-VBA3), the RFP of PrVmAIP showed the presence of mutations within the RE sites studied. Both PrV-VBA1 and PrV-VBA2 appeared to be closely related but their RFPs differed from PrV-VBA3. Significant differences either in the number, size or migrations of the DNA fragments could also be detected in the BUdR resistant strains. Even though different features of cytopathic effect (GPE) were observed in the IUdR resistant PrVs, the RFP findings remained identical. The PrVs studied showed considerable differences from the reference PrV (PrV-CD). PMID- 11942087 TI - Conceptual blockbusters: creative idea generation techniques for health administrators. PMID- 11942088 TI - The manager and pain control for patients. AB - Every day seems to bring new demands for health services managers, even though none of the existing expectations seems to end. But, an effective pain management program is basic to meeting patient needs and is driven by patients' judgments and demands. It remains for HSOs and their managers to determine the means by which patients' judgments and demands will be met. PMID- 11942089 TI - Verbal abuse of female nurses: an American medical form of gender apartheid? PMID- 11942090 TI - Sexual harassment in healthcare: strategies for employers. PMID- 11942091 TI - The lesson of the slime mold. PMID- 11942092 TI - Surfing the edge of chaos. Interview by Joe Flower and Patrice Guillaume. PMID- 11942093 TI - New climate for foreign workers. PMID- 11942094 TI - Surviving mergers & acquisitions. AB - Mergers and acquisitions are never easy to implement. The health care landscape is a minefield of failed mergers and uneasy alliances generating great turmoil and pain. But some mergers have been successful, creating health systems that benefit the communities they serve. Five prominent leaders offer their advice on minimizing the difficulties of M&As. PMID- 11942095 TI - Winning over the medical staff. By creating a Physician Advisory Group, East Jefferson gained the trust of its physician staff. PMID- 11942096 TI - Planning for the unknown. PMID- 11942097 TI - C.A.M. Complementary & alternative medicine. JCAHO includes CAM therapy. PMID- 11942098 TI - Best practices. A scorecard for players. PMID- 11942099 TI - Eat dessert first. Nothing is certain, especially in health care. PMID- 11942100 TI - Florida nursing students: our nursing future. PMID- 11942101 TI - New initiative on geriatric nursing education issues formal report outlining goals and strategies. PMID- 11942102 TI - Nursing malpractice insurance: individual insurance policy purchase. PMID- 11942103 TI - Life in a pill bottle: the experience of taking HIV medications. PMID- 11942105 TI - Patient safety--new falls prevention initiative. PMID- 11942104 TI - Florida Nurses Association: clearing the myths and misconceptions. PMID- 11942106 TI - Vibrio vulnificus and the high risk. Patient: a potentially fatal disease. PMID- 11942107 TI - Quality end-of-life care across all settings: the specialty of hospice palliative care. PMID- 11942108 TI - Tobacco-free Florida. PMID- 11942109 TI - Assessing for smallpox. PMID- 11942110 TI - Communication: it can make us or break us! PMID- 11942111 TI - Our voices together can make a difference. PMID- 11942112 TI - Physical security and HIPAA compliance: what two leading health systems are doing. PMID- 11942113 TI - Weronik: JCAHO, 9/11 require new definition of--and greater protection for--VIP patients. PMID- 11942114 TI - Employee screening today: the information is there, but be careful how you use it. AB - Doing a good job of conducting thorough reference checks and background investigations of job applicants has become increasingly important in healthcare in preventing personnel-related crime and the legal consequences of negligence in hiring people who commit such crimes. However, there are many obstacles to obtaining information on applicants. In this report, we'll explore in depth the current state of background checking as described by an official of a company which provides background checking services, as well as the latest in legal practices from a leading attorney in the field. PMID- 11942115 TI - Consent form wins case for hospital in employee defamation suit. PMID- 11942116 TI - Frequencies of feline blood types in Hungary. AB - Feline blood group determination is done as a routine diagnostic method in numerous countries. Blood transfusion reactions and feline neonatal isoerythrolysis (FNI) can be avoided with the identification of different feline blood groups. The present study is the first investigation in Hungary during which 100 cats have been examined from all over the country. These cats were out of six breeds: European domestic shorthair, Persian mix, Persian, Abyssinian, Siamese and British shorthair. In the Hungarian feline population European domestic shorthair are most common but other breeds also occur. European domestic shorthair, Persian mix, Abyssianian, Siamese and British shorthair individuals all belonged to blood type A (100%). Blood type B was found very rarely and only in Persian cats. One-third of the Persian cats were categorised into blood type B, whilst type AB was not found during the study. PMID- 11942117 TI - Testing of a Chemiluminescence Enzyme Immunoassay for selective detection of E. Coli O157 from ground beef samples. AB - The aim of this study was to evaluate a Chemiluminescence Enzyme Immunoassay (CLIA) developed for the detection of E. coli O157:H7, using different E. coli O157 serotypes. The sensitivity and specificity of the kit were determined from the tenfold dilutions of the 24-hour broth cultures of the test strains. According to the results obtained in this trial, the sensitivity of the kit is 10(3)-10(4) cells ml-1, and it is specific for E. coli O157. Twenty-five g ground raw beef samples were prepared and inoculated with E. coli O157:H7 at different CFU g-1. The samples were incubated in 225 ml of modified E. coli broth with novobiocin (mEC + n) at 42 degrees C for 4 h and the immunoassays were performed following the instructions of the manufacturer. According to the results obtained by the CLIA test 10(1)-10(2) E. coli O157 g-1 can be detected from the sample. So this kit seems to be suitable for screening the samples before selective cultivation of E. coli O157:H7. PMID- 11942118 TI - Alteration of body composition in rats: effect of organic chromium and L carnitine. AB - A growth trial was carried out to test the effect of organic, trivalent chromium and L-carnitine on the body composition of growing rats. At the same time, an evaluation of different measurement methods (weight of epididymal fat pad, adipocyte morphometry, total body electrical conductivity) was performed. Outbred Wistar rats of 30 days of age were fed diets of different (0, 10 and 20%) protein level. The diets were supplemented with 4 mg/kg Cr as chromium nicotinate, and 100 mg/kg L-carnitine. The experimental feeding lasted 15 days, after a 5-day long adjustment period. It was found that Cr addition increased feed intake. Both treatments caused changes in body composition, increasing fat and protein deposition. Organic chromium had no effect at either protein level, while L carnitine improved the protein retention only at an optimum (20%) protein supply. No statistically significant correlation was found between total body electrical conductivity (TOBEC) and body composition, which could be attributed to the great individual differences. A close correlation was found among total body fat percentage, weight of epididymal fat pad and the adipocyte surface. The data suggest that there is an interaction between dietary protein supply and the effect of repartitioning agents. PMID- 11942119 TI - Influence of feeding intensity on corporeal development, body composition and sexual maturity in female rabbits. AB - Twenty-six 6-week old female New Zealand White rabbits were divided into two groups: ad libitum (AL) and 70% restricted (RS) feeding. At the beginning of the experiment the liveweights were practically the same: 0.99 +/- 0.08 vs. 1.01 +/- 0.08 kg in group AL and RS, respectively. At 18 weeks of age the body weight of Group RS rabbits was 84.7% (3.14 +/- 0.24 kg) of the group AL (3.71 +/- 0.31 kg). The apparent digestibility of crude protein was significantly (p < 0.001) higher in Group RS than in Group AL (76.5 +/- 1.4 vs. 73.0 +/- 2.7%). The daily water consumption was significantly (p < 0.05) higher (3.5 ml/g DM intake) in Group RS as compared to Group AL (1.9 ml/g DM). Since the average body weight in Group RS at 18 weeks was the same as that of Group AL at 14 weeks of age (3.14 kg), the comparison of the live body measures and indices was based on these ages. Animals fed ad libitum or restricted show no differences at the defined age in most live body measurements except in heart girth and rump width, which were significantly (p < 0.05) shorter in Group AL than in Group RS (29.3 +/- 0.8, 5.7 +/- 0.5 and 30.7 +/- 1.0, 6.2 +/- 0.3 cm, respectively). Body capacity was significantly (p < 0.05) smaller in Group AL, as the head capacity-body capacity proportion was significantly (p < 0.05) higher in Group AL than in Group RS (1653.1 +/- 134.5, 22.0 +/- 2.5 and 1913.7 +/- 168.7, 17.7 +/- 1.9, respectively). Due to restricted feeding the growth of the head proved to be less intensive than that of the body at the same body weight. The body in these does tended to be wider. Since the head in comparison to length or capacity of the body was smaller in does fed 70% of ad libitum, it could be concluded that the development of body parts of restricted-fed does was unequal (allometric growth). The effect of feed restriction reflected in lower dry matter and fat, and a higher ash and protein content both in total body and in dry matter composition of rabbits at 18 weeks of age. Restricted feeding delayed sexual maturity (69 vs. 92% of rabbits) with later starting ovarian activity, weaker ovarian responsiveness, and a smaller number of tertiary follicles on the ovarian surface. PMID- 11942120 TI - Erythrocyte osmotic fragility test as the measure of defence against free radicals in rabbits of different age. AB - Peroxidation of the unsaturated bonds of membrane lipids increases fragility and cellular lysis of red blood cells. Erythrocyte susceptibility to the free radicals (peroxyl radicals) generated in vitro by 2,2'-azo-bis(2-amidinopropane) hydrochloride (AAPH) was evaluated and expressed as 50% maximal haemolysis time (HT50) in 3 groups of rabbits of different age. Erythrocytes of 1.5-month-old rabbits were more sensitive to free radicals than those of 3.5- and 6-month-old ones. In the three groups, significant negative correlation (r = -0.8 to -0.98) between the lipid peroxidation rate (thiobarbituric acid reactive substances; TBARS concentration) in blood plasma and the erythrocyte resistance to free radicals was found. This result suggests that the plasma antioxidant defence system is interrelated with that of the red blood cells and that the erythrocytes can be a good model for studies of oxidative stress. The simple haemolysis test reflecting the free radical defence can be useful for evaluating the antioxidant properties of various compounds. PMID- 11942121 TI - A simple way to reduce heat stress in laying hens as judged by egg laying, body weight gain and biochemical parameters. AB - This study was designed to test the effects of feed withdrawal and darkening on the performance, triiodothyronine (T3), thyroxine (T4), thyroid-stimulating hormone (TSH), adrenocorticotropic hormone (ACTH), and some blood serum metabolite and mineral concentrations of laying hens reared at high ambient temperatures ranging from 25 to 35 degrees C. Ninety, 16-week-old hens (Ross Brown) were divided into 3 groups, 30 hens each. The first group was used as control. Hens in the second group (feed withdrawal) were subjected to feed removal from 14:00 to 18:00, and hens in the third group (darkening) were subjected to light restriction from 14:00 to 18:00 using black curtains. Liveweight, feed intake, and egg production were higher (P < 0.01) in the feed withdrawal and darkening groups, particularly in the darkening group, than in the control. Water intake was higher in the control group compared with the feed withdrawal and darkening groups (P < 0.01). T3, T4, and TSH concentrations in the serum were higher (P < 0.01), whereas ACTH serum concentration was lower (P < 0.01) in the feed withdrawal and darkening groups compared with the control. The haematocrit was higher in the feed withdrawal and darkening groups compared with the control (P < 0.01). Darkening and feed withdrawal treatments increased serum glucose, urea-N, uric acid, albumin, triglyceride, cholesterol, Ca, P, Na, and K concentrations, also the activities of amylase and alkaline phosphatase, but did not influence the activities of serum glutamic-oxaloacetic transaminase (SGOT) and serum glutamic pyruvic transaminase (SGPT). The present study found that feed withdrawal and darkening, particularly darkening, at high temperatures during the summer months offer a good management practice to reduce heat stress related depression in feed intake and egg production in laying hens. PMID- 11942122 TI - Studies on factors affecting superovulation and embryo transfer in Hungarian merino ewes. AB - The objectives of this study were (a) to assess the ovulatory response and embryo production of Hungarian Merino ewes after superovulation, (b) to investigate the factors influencing the efficiency of embryo transfer (ET) in Hungarian Merino ewes, (c) to compare the results of two ovarian stimulation protocols (PMSG and PMSG + FSH treatment) in Hungarian Merino ewes, and (d) to study how superovulation, laparoscopic insemination and surgical embryo retrieval (ER) affect the subsequent reproduction of Hungarian Merino donor females after an ET programme. There was no significant difference between the ovarian stimulation protocols in the ratio of donor ewes responding to superovulation nor in the average number of corpora lutea. However, the number of transferable embryos recovered per donor ewe was higher in the PMSG + FSH group. The proportion of transferable embryos, unfertilized oocytes and degenerated embryos did not differ between the treatment protocols. The total pregnancy rate was 53.4% (179/335). Neither the developmental stage of the embryo nor the number of transferred embryos affect the implantation of embryos. However, the increased number of transferred embryos positively influenced the pregnancy rate. No difference was found in the pregnancy rate between synchronised and non-synchronised groups of recipients. Thirty-six out of 45 donor ewes (80%) became pregnant within one year after the ET programme, indicating that ovarian stimulation and surgical ER did not affect adversely their reproduction. PMID- 11942123 TI - Ovarian stimulation and ultrasound-guided oocyte retrieval in baboon (Papio Cynocephalus Anubis) during pituitary suppression with a GnRH agonist. AB - The objective of this study was to investigate whether baboon females respond to an ovarian stimulation protocol incorporating pituitary suppression with a GnRH agonist (GnRHa) and either highly purified human FSH (hphFSH) or recombinant human FSH (rhFSH) with follicular development and oocyte maturation. A modified human ovulation induction protocol was applied to 5 adult female baboons with a history of regular menstrual cycles (33-34 days). A long-acting GnRHa implant containing goserelin acetate was placed subcutaneously (s.c.) on Days 22-24 of their menstrual cycle. Concentrations of serum oestradiol (E2), progesterone (P4) and human FSH were obtained by ELISA. Menses occurred approximately 10 days after GnRHa implantation. Daily hphFSH or rhFSH (75 IU i.m.) treatments were started approximately 10 days following menses. When the majority of follicles were > or = 5 mm in diameter and the E2 levels had reached a maximum, hCG (2000 IU i.m.) was administered to induce final maturation of oocytes and ovulation. Thirty to 34 h after hCG administration, transabdominal follicular aspiration was performed using a variable frequency transvaginal transducer with ultrasound. A total of 71 oocytes were collected from 4 animals (average: 17). The meiotic maturity of oocytes was evaluated 3 h after retrieval. Ninety-one percent of oocytes were in metaphase 2 and of grades I and II which are appropriate for in vitro insemination. PMID- 11942124 TI - Endoscopy and autopsy follow-up of biodegradable oesophageal anastomoses in dogs. AB - The biofragmentable Anastomosis Ring (BAR) is a mechanical device composed of absorbable material and creates an inverting, atraumatic compressive anastomosis with no foreign material at the anastomotic site after healing. The aim of the present experimental study was to assess the safety of oesophagoscopy in early days after oesophageal anastomoses performed with the BAR and to follow-up the healing of BAR anastomoses by in vivo endoscopy and autopsy examination. Thirty mongrel dogs divided into subgroups according to the time-points of endoscopy and autopsy (4th, 7th, 14th, 28th day) were used. There was no significant difference in the healing of anastomoses performed under or above the tracheal bifurcation. Pleural adhesions helped to cover and seal small subclinical leaks. The mortality was 13.3% (4 dogs) and the overall leakage rate 14.3%. We looked for bleeding, haematoma, erosion, ulceration and granulation tissue in the anastomosis. Due to the high mechanical strength of these anastomoses, oesophagoscopy was a safe, easy and feasible method for follow-up BAR intrathoracic anastomoses, with no significant difference between the number of lesions found with endoscopy as compared to the autopsy data. The overall sensitivity of oesophagoscopy to discover mucosal lesions was 73.1%. Endoscopy had no complications, therefore it is a useful method of follow-up and may help predict the normal or compromised healing of oesophageal anastomoses. PMID- 11942125 TI - Decreasing half-life of dieldrin in egg yolk following a single oral administration of aldrin to laying hens. AB - Laying hens were treated orally with a single dose of aldrin (AD) 1 mg/kg body weight. Concentrations (microgram/g) of AD or its epoxide (= dieldrin, DD) in the yolk of eggs laid for 21 days after AD treatment were determined by normal-phase high-performance liquid chromatography. The limits of determination were 0.02 microgram/g for AD and 0.03 microgram/g for DD, respectively. After AD treatment, although the low levels of AD (mean 0.02-0.03 microgram/g) were observed only during a three-day period (from 4th to 6th days), DD (mean 0.15 microgram/g) was found already on the 2nd day, indicating that the epoxidation of AD to DD in the hen's body is rapid. The highest level of DD (mean 0.40 microgram/g) was detected on the 6th day, and then DD levels decreased slowly and were detected up to the 21st day. In this decreasing phase, the half-life of DD in the yolk was estimated to be 25.6 days with a 95% confidence interval from 22.7 to 29.4 days. PMID- 11942127 TI - [Hot topic of the new year: is nursing a career profession--or not?]. PMID- 11942126 TI - EHV-1 gene63 is not essential for in vivo replication in horses and mice, nor does it affect reactivation in the horse: short communication. AB - The aim of this study was to investigate the role of immediate early gene (gene63) in the pathogenesis of equine herpesvirus 1 (EHV-1) acute and latent infections in equine and murine models. EHV-1 gene63 mutant virus (g63mut) along with EHV-1 (Ab4) was used for intracerebral and intranasal infection of 3 and 17 day-old mice. Both viruses were recovered at the same frequency from tissues after infection. Two Welsh ponies were infected via the intranasal route with each of the viruses. Acute infection was monitored by virus isolation from nasal swabs and peripheral blood leukocytes. Six weeks post infection, peripheral blood leukocytes were taken from ponies and in vitro reactivation was positive for both viruses. At autopsy, both viruses were isolated by co-cultivation from bronchial and submandibular lymph nodes. These findings indicate that the mutation of EHV-1 gene63 does not play a role in the establishment and reactivation from latency. PMID- 11942128 TI - [Career path in nursing]. PMID- 11942129 TI - [Career in nursing in connection with an empirical inquiry study]. PMID- 11942130 TI - [Health and social environment]. PMID- 11942131 TI - [Nursing and health-/social policy]. PMID- 11942132 TI - [What is public health?]. PMID- 11942133 TI - [AIDS--observations between sexuality and hand disinfection]. PMID- 11942134 TI - [Demographic trends, increased need for nursing care, educational reform and career choice. The central question for the future: when will there be the next nursing nursing shortage?]. PMID- 11942135 TI - [Living and health vs. freedom and human dignity]. PMID- 11942136 TI - [Sensory deprivation in advanced age and nursing care in inpatient nursing facilities]. PMID- 11942138 TI - [Violence and aggression in professional nursing care... silence means collaboration... ]. PMID- 11942137 TI - [Women and violence]. PMID- 11942139 TI - [Violence against children... ]. PMID- 11942140 TI - [A survey on fatal work accidents based on Mortality Registry data: results of the Tuscany study on INAIL and RMR cases in the period 1992-2996]. AB - Work-related deaths are important "sentinel events" of unsuccessful prevention. In Italy the most exhaustive source of such events is the National Fund for Occupational Diseases (INAIL), but the amount of cases from this source seems to be underestimated due to the fact that it refers only to cases occurred to subjects insured by the Fund. A previous survey estimated the real amount of work related deaths 10-20% higher than that quantified by INAIL. This study evaluated the contribute of the two most important sources (INAIL and the Regional Mortality Registry of Tuscany-RMR) in estimating the number of these cases in Tuscany in the period 1992-96. Cases were identified from each source, and then it was applied a capture-recapture method to size the cases from work-related accident different from road accidents. RMR appeared to be the most exhaustive source with 72.3% completeness versus 56.4% completeness of INAIL source. Nevertheless the last one must be considered the primary source, more specific and accurate, and since few years also timely, than any other one. Work-related deaths from road accident represent 35.9% of INAIL cases but they are difficult to be identified from RMR and were not considered in this study. In conclusion the mortality data should be used for an epidemiologic surveillance system on work-related deaths not due to road accident in order to identify cases occurred to subjects not insured by INAIL (and therefore not defined by the Fund). These deaths are also important in terms of public health. Cases identified only from RMR, occurred in Tuscany in 1992-96, were 155: the vast majority occurred to farmers (mainly pensioners, and due to caterpillar upsetting), to bricklayers, to railway workers, to soldiers and to entrepreneurs. PMID- 11942141 TI - [Health impact of exposure to environmental tobacco smoke in Italy]. AB - Household and workplace exposure to environmental tobacco smoke (ETS) is associated with various deleterious health effects. We evaluated the health impact of ETS in Italy. We considered only those health conditions for which systematic quantitative reviews were available. We used relative risks from the reviews while data on the proportion of the Italian population exposed to ETS were derived from national statistics or from ad hoc studies. A total of 2.033 newborns per year are of low birth weight (< 2500 grams) in Italy (7.9%) because of maternal exposure to ETS during pregnancy. There are 87 sudden infant deaths (16.9%) due to post natal maternal active smoking. 21.3% of acute respiratory infections during the first two years of life are due to parental smoking (about 77.000 children); 27.000 cases of asthma (9.1%), 48.000 cases of chronic respiratory symptoms, 64.000 cases of middle ear effusions among children and adolescents are due to parental smoking exposure. The annual estimates of mortality from lung cancer and ischemic heart disease due to ETS exposure from the spouse are 221 and 1896, respectively. Workplace exposure is associated with 324 lung cancer deaths and 235 deaths from ischemic heart disease. Although these estimate are based on several assumptions and they have large uncertainties, the results underline the need and the urgency of public health measures to reduce ETS exposure in the household and in the workplaces in Italy. PMID- 11942142 TI - [Speed limits. A letter to the Minister of Transportation Pietro Lunardi]. PMID- 11942143 TI - [Why shouldn't Italy host the Centenary Congress of the International Commission on Occupational Health (ICOH) in 2006?]. PMID- 11942144 TI - [Real-time measurement of indoor particulate matter originating from environmental tobacco smoke: a pilot study]. AB - Short-term measurement of suspended particulate matter has been recently made possible since the release of laser-operating portable instruments. Data of a pilot study of field evaluation of environmental tobacco smoke (ETS) with a portable instrument are reported. We analysed the concentrations of total suspended particle (TSP) and of the fine particles PM10, PM7, PM2.5 and PM1 released indoor from a single cigarette, and their levels inside smoking- and non smoking-areas of a restaurant. The results indicate that ETS creates high level indoor particulate pollution, with concentrations of PM10 exceeding air quality standards. This kind of field evaluation could allow a more careful assessing of short-term exposure to ETS and its relevance to public health. PMID- 11942145 TI - [Environmental tobacco smoke (ETS) at the workplace: considerations about a survey carried out in a pharmaceutical industry]. AB - A survey was carried out in a pharmaceutical industry, in order to know the exposure to environmental tobacco smoke (ETS) in the workplace, and attitudes of workers toward workplace smoking restrictions and bans. In the three plants of the industry (Bergamo, Milan, Reggello-Florence) workers were exposed to ETS in every workplace areas, except for the production area, even if there were no smoking warning notices in the plants. Non-smokers were more favorable than smokers about the introduction of workplace restrictions on smoking. In order to decrease exposure to ETS, it would be desirable to develop workplace smoking restrictions or bans, effective and low-cost interventions. PMID- 11942146 TI - [The Italian epidemiology in the new millennium. Considerations for the analysis of the role]. PMID- 11942147 TI - [A practical approach to reduction of traffic accidents]. PMID- 11942148 TI - [One Euro against tobacco. To prevent smoking and to help people who want to quit]. PMID- 11942149 TI - [Occupational accidents: a phenomenon needing in-depth inquiry]. PMID- 11942150 TI - [Focus on radon]. PMID- 11942152 TI - Madeleine Leininger on transcultural nursing and culturally competent care. Interview by Mary Agnes Seisser. PMID- 11942151 TI - Medication error identification is a key to prevention: a performance improvement approach. AB - The leadership of Memorial Hospital of Carbondale sponsored an interdisciplinary continuous quality improvement team effort to address medication errors. The need for increased awareness, documentation, and reporting of medication errors throughout the organization was identified as a priority through risk assessment. Ineffective reporting of medication errors occurred in all medication distribution categories: prescribing, transcribing, dispensing, and administration. Improvement interventions focused on building an awareness of the occurrence of medication errors through identification and on the reporting of those errors to the risk management department. PMID- 11942153 TI - The healthcare quality profession: changing and changing fast. PMID- 11942154 TI - Guidelines for improving nutritional delivery in the intensive care unit. AB - Optimizing nutritional delivery in the intensive care unit (ICU) continues to be a challenge. Nutritional guidelines were developed at a metropolitan Level I trauma center as an institutional response to ensure the timeliness of patient evaluation, initiation of therapy, and attainment of goal therapy. A post implementation review of 525 consecutive ICU patients revealed that the guidelines enabled the staff to evaluate 86% of all ICU patients and initiate appropriate therapy in 68% of them within 48 hours of admission. Goal therapy was achieved in more than 90% of patients within 72 hours. The establishment of nutritional guidelines is an integral step to improving nutritional therapy in the ICU. PMID- 11942155 TI - Determinants of patient satisfaction in a military teaching hospital. AB - The purpose of this study was to determine the aspects of hospital services that are most likely to affect patient satisfaction in a military teaching hospital in Turkey. Although there have been many studies on patient satisfaction in Turkey and other countries, few studies have been done in military hospitals. A patient satisfaction questionnaire using a 4-point Likert scale was mailed to 500 patients after discharge, and 316 questionnaires were returned. The findings indicated that satisfaction with physician, nursing, physical plant, and food services were the main determinants of overall satisfaction with the hospital. The type of clinic in which the patients stayed also was an important determinant. The effect of patients' demographic characteristics on overall satisfaction with the hospital was also examined, and only lower education level was a statistically significant determinant. PMID- 11942156 TI - Integrating service excellence in a CHF clinical pathway pilot project. AB - The complex dynamics of the current healthcare environment require healthcare delivery systems to become cost effective and quality driven. Educated healthcare consumers expect superior service and timely responses to their needs. For one healthcare system, customer expectations were an integral part of designing, implementing, and measuring the service components of congestive heart failure pathway outcomes. Service excellence can influence overall clinical outcomes when measured by consumer awareness and patient satisfaction. The inclusion of service excellence as an intrinsic piece of the organizational strategic plan laid the groundwork for this integrated pilot project. PMID- 11942158 TI - Harmonizing care, costs, and quality: turning around the negative 1990s. PMID- 11942157 TI - Inpatient psychiatry incident review management: Part I. Special issues. AB - Incident review (IR) is a process for achieving a better understanding of problems related to the health and safety of clients and the integrity of healthcare facilities. The purpose of IR is to identify and document adverse incidents through fact-finding. Incidents may vary in both type and levels of seriousness. Through careful documentation, investigation, and committee review, patterns of healthcare delivery can be discerned. The goal of IR is the development of systemic performance improvements in healthcare delivery, which may serve to anticipate and deter the recurrence of similar incidents. IR ensures that the quality and performance of healthcare delivery in the facility continue to be improved by both administrative and clinical staff members. This article examines IR in the context of the inpatient psychiatry unit. PMID- 11942159 TI - Developing a prioritization matrix for improving breast care--a complex project. AB - Prioritization is an ongoing challenge for quality management professionals. A previous companion Brief Report described a tool for prioritizing improvement projects (Pelletier, Beaudin, and van Leeuwen, 1999). This tool provides a framework for prioritizing opportunities/solutions for major institutional improvement projects. By separating and categorizing opportunities and solutions and then weighing their importance and impact, the organization can graphically evaluate and then select interventions to initiate. PMID- 11942160 TI - Impact on quality and patient safety: the new shortage of healthcare professionals. PMID- 11942161 TI - Subject: James Bagian on patient safety initiatives. PMID- 11942162 TI - The relationship between continuous quality improvement and research. PMID- 11942164 TI - [New perspectives in immuno-mediated blood diseases]. AB - Immune-mediated blood diseases are organ-specific autoimmune disorders where one or more blood cell lineages are the target of the autoimmune process. They are classically divided into peripheral immune-mediated blood diseases when the immune attack occurs outside the bone marrow and autoimmune myelopathies. Overall they are the prototype of diseases that bridge immunology and hematology including blood malignancies. They also provide a fertile ground for new modalities of immunosuppression. The investigations of platelet autoimmunity as a model of peripheral immune-mediated blood diseases have confirmed that platelet disorders hover between hemorrhage and thrombosis and have led to develop the concept of autoimmune thrombophilia. PMID- 11942163 TI - [Antibiotic resistance and community-acquired infections]. AB - The selective pressure of extended broad-spectrum antibiotic administration is leading to a progressively growing selection of common gram-positive and gram negative organisms which show unpredictable in vitro antimicrobial susceptibility levels to current first-line therapeutic choices. This phenomenon is examined from an epidemiological, microbiological, therapeutical, and pharmacoeconomic point of view, focusing our attention on the most common community-acquired infectious diseases interesting upper and lower airways, genito-urinary and gastrointestinal tract, skin and soft tissues. A broad spectrum of variables is responsible for this ever changing scenario, whose solution cannot be entrusted to the development of new compounds, but it first relies on a rational and judicious administration of existing molecules, and an expanded medical and patient interaction aimed at avoiding overprescription of antimicrobial agents. PMID- 11942165 TI - [Community-acquired pneumonia: outcomes and costs]. AB - The number of patients admitted with community acquired pneumonias (CAP) varies greatly from one hospital to another. Prognostic models for CAP can help physicians decide which cases to treat on an outpatients basis. Our aims were: a) to validate a model for predicting low-risk CAP, and b) to estimate savings that would have resulted if the low-risk patients identified by the model had been treated at home rather than in hospital. The prediction rule of Fine et al. was used to classify retrospectively 260 CAP patients. Mortality in each category was compared with the mortality predicted. Patients in the lowest risk categories were considered to have been inappropriately admitted. The predictive model used has been found useful for identifying patients at very low-risk of dying from CAP. Application of the model can lead to savings. PMID- 11942166 TI - [Transthoracic echocardiography in the diagnosis of pulmonary embolism]. AB - Transthoracic echocardiography is advocated by some as a useful diagnostic test for patients with suspected pulmonary embolism (PE), but its diagnostic accuracy is unknown. Recently, the sensitivity and specificity of transthoracic echocardiography in the diagnosis of PE was determined prospectively in unselected patients using pulmonary angiography as a reference diagnostic standard. Echocardiographic criteria for diagnosing PE included the presence of any two of three abnormalities: right ventricular (RV) end-diastolic diameter > 27 mm (without RV wall hypertrophy), tricuspid regurgitation velocity > 2.7 m/s, or RV hypokinesis. These echocardiographic diagnostic criteria for PE yielded a sensitivity of 56% and a specificity of 90%. Because of its inherent poor sensitivity, transthoracic echocardiography should not be used as a routine test to screen patients for suspected PE. PMID- 11942167 TI - [Bulbar paralysis and flaccid tetraparesis in thyrotoxicosis]. AB - We describe the case of an aged patient, presenting an acute onset of flaccid paralysis of the limbs and bulbar weakness, complicated by ab ingestis pneumonia. These neurological findings were the first signs of a thyrotoxic crisis and showed a progressive regression together with the healing of the hyperthyroidism obtained, within two months, utilizing thyroid inhibitors. At the onset, a marked hypokalaemia was present, making possible the hypothesis of a thyrotoxic periodic paralysis. The hypokalaemia was corrected after many infusions of KCl i.v., without obtaining significant modifications of the neurological picture. Taking into consideration the evolution and the results of the instrumental procedures (EMG, MRI) we believe this a rare case of thyrotoxic acute myopathy, with bulbar involvement, casually associated to hypokalaemia. PMID- 11942168 TI - [Hyperglycemia and vascular damage role of oxidative stress]. AB - Diabetes mellitus is an important risk factor for cardiovascular disease and hyperglycaemia is being more and more acknowledged as a mediator of vessel damage. It is becoming increasingly clear that hyperglycaemia involves a number of different parameters (acute, chronic, and post-prandial hyperglycaemia), all of which contribute to the vascular damage caused by high glucose levels. High glucose levels are associated with the non-enzymatic glycation of both extra- and intracellular proteins, the accumulation of sorbitol via the aldose-reductase pathway, the activation of protein kinase C isoforms, and the reduced bioavailability of nitric oxide. The generation of reactive oxygen species may be a common downstream mechanism by means of which the multiple by-products of glucose exert their adverse effects on blood vessels. PMID- 11942169 TI - [Adjuvant treatment of prostatic carcinoma after primary therapy. The "Early Prostate Cancer" Program]. PMID- 11942170 TI - [Adjuvant hormone therapy in prostatic cancer: after treatment with curative intent. The viewpoint of the urologist]. PMID- 11942171 TI - [Efficacy of bicalutamide in adjuvant treatment of prostatic carcinoma after primary therapy with curative aim. The viewpoint of the surgeon]. PMID- 11942172 TI - [Clinical implications of rheology in cardiovascular diseases]. AB - Although a close relationship between haemorrheological disorders and thrombogenesis has been known for a long time, only the recent introduction of newly developed analytical techniques has allowed a precise assessment of the main haemorrheological parameters. The employment of these techniques in routine clinical practice has enabled, for instance, a more accurate investigation of the pathogenetic triggers underlying numerous thrombotic disorders affecting the cardiovascular system. A survey of the most exhaustive prospective studies shows that some haemorrheological parameters (e.g. plasma and blood viscosity) as well as their main determining factors (e.g. haematocrit and fibrinogen) are well defined independent cardiovascular risk factors. Nonetheless, further large scale studies are required to adequately dissect the relative contribution of both haemorrheological alterations and other thrombophilic factors in the pathogenesis of different cardiovascular diseases related to thrombotic disorders. PMID- 11942174 TI - [Endothelial acetylcholine]. AB - A fraction of the vascular endothelial cells may synthetised and release acetylcholine. This release is enhanced by mechanical stimuli such as shear stress or increased blood flow. The physiological role of this release of acetylcholine is explained taking the vasodilatory response to mental stress of blood vessels in skeletal muscles as an example. PMID- 11942173 TI - [Piperacillin-tazobactam in the treatment of severe nosocomial infections]. AB - Epidemiology and clinical manifestations of nosocomial infections have been progressively evolving through last three decades. New pathogens, often associated to multiple antibiotic-resistances, are now among the leading etiologies in a patient population mainly represented by individuals debilitated by serious underlying diseases, immunosuppressive treatments and multiple diagnostic and therapeutic invasive procedures. Under these circumstances, when an infectious complication develops, prompt initiation of empiric antibiotic therapy, based on general and/or local epidemiology of nosocomial infections, may be crucial for survival of more critically ill patients. Based on recent data, the antibiotics of choice for treatment of nosocomial infections should have not only a bactericidal activity and a broad antibacterial spectrum, but also poor propensity to induce antibiotic-resistance and a satisfactory pharmacoeconomy profile. After a detailed description of the above mentioned context, the pre eminent role of piperacillin-tazobactam in the treatment of nosocomial infections is reviewed and discussed. PMID- 11942175 TI - [War and medicine in a culture of peace. 4. Synopsis of nuclear weapons]. AB - The nuclear weapons have a terrifying power of devastation. A civilian nuclear accident has its own dramatic consequences. Both situations are, however, totally distinct with regard to medical consequences. The Gulf war and Balkan war syndromes might be in part caused by uranium used in conventional amunitions. The risk of nuclear terrorism is an emerging eventuality. PMID- 11942176 TI - [How I explore ... the risk of a patient developing type 2 diabetes]. AB - Both the prevalence and the incidence of type 2 diabetes are increasing rapidly. Effective prevention measures, including lifestyle or drug prescription, have been recently reported. It is thus important to detect at risk individuals in order to provide appropriate diet and exercise recommendations or even pharmacological treatment. We summarize the most useful indices based on anamnesis, clinical examination and biological assays that can help to detect subjects at high risk of progression towards type 2 diabetes. PMID- 11942177 TI - [Pharma clinics. Medication of the month. Imiquimod (Aldara): an immunomodulator for the skin]. AB - Imiquimod is the first drug of a new therapeutic class encompassing cutaneous immune response modifiers. This molecule induces the synthesis and release of macrophage-related cytokines. It boosts some local immune responses. These properties are used to treat cutaneous viral condylomas. Other prospective therapeutic goals are presently scrutinized. PMID- 11942178 TI - [Clinical study of the month. Adjuvant radio-chemotherapy and chemotherapy following curative resection of pancreatic cancer: results of the randomized trial ESPAC-1]. AB - The prognosis of pancreatic adenocarcinoma remains poor, with a 5-year survival rate lower than 5%. Resection, the gold standard treatment, can be performed in less than 15% of patients. Following surgery, the median survival is 12 months for the most favourable cancer patients. Adjuvant treatment have attempted to improve results. However, chemotherapy, radiotherapy and multimodal treatments don't have demonstrated a clear advantage in controlled trials. We will discuss results of the current trials in this topic. The randomised trial of the European Study Group for Pancreatic Cancer (ESPAC) recently published in the Lancet revealed a potential benefit of adjuvant chemotherapy. A critical analysis of the publication showed, however, that definitive conclusions of this trial must be interpreted with caution. PMID- 11942179 TI - [Image of the month. Preauricular tumor]. PMID- 11942180 TI - [Pharma clinics. How I treat ... recurrent herpes labialis]. AB - Recurrent herpes labialis is a frequent disorder. It occurs following the reactivation of the Herpesvirus (HSV1 and more rarely HSV2) inside the Gasser ganglion. Treatment and prevention of recurrent labial herpes are targeted by specific antiviral agents. Spectacular benefits obtained in the immunocompromised patients are less convincing in otherwise healthy subjects. Other prospective ways of therapy are under consideration, including lipopeptides and physical means aiming at modifying the cutaneous pH. PMID- 11942181 TI - [Clinical case of the month. An unusual cause of bronchiectasis: primary ciliary dyskinesia]. AB - We report a case of bronchiectasis due to primary ciliary dyskinesia. PMID- 11942182 TI - [Do calcium antagonists have a first-line role in the treatment of arterial hypertension?]. AB - During the nineties, a controversy was noted in the literature about the potential risk of using calcium antagonists in the treatment of hypertension. Several recent studies, such as the INSIGHT's one have not confirmed this provided that one uses long-acting molecule. But the real beneficial effect due to the different antihypertensive classes is mainly linked to the magnitude of the blood pressure lowering effect. PMID- 11942183 TI - [Diagnosis of internal carotid artery dissection. Two case reports]. AB - Two cases of acute internal carotid dissection are presented. Typical symptoms, pathogeny and imaging features are reviewed. Magnetic Resonance is actually the best technique for the diagnosis of internal carotid artery dissection, which should be searched in young patients presenting neurologic and cervico-facial symptoms. PMID- 11942184 TI - [Suicide in the elderly]. AB - This article shows the different aspects of suicide in older people. The numerical ratio between suicide and suicide attempt decreases with age. Among female, under 25 years old, this ratio is 1 suicide for 160 suicide attempts. Among male, the margin is smaller because we have 1 suicide for 1.2 suicide attempts. Suicide rates among the elderly are the highest of any age group. Widowhood, social isolation, somatic illness, and major depression are the most important risk factors. Among the elderly, suicide is more radical. Hanging and drowning represent the principals modes of suicide. The prevention is very essential but difficult because of a great number of various clinical situations associated with suicidal risk. A substantial proportion of late life suicides have contact with their primary care providers prior to their death. This offers a unique role for general practitioners who should be well prepared to diagnose and treat depression in older patients. PMID- 11942185 TI - [Evaluation of tetanus vaccine coverage in rural society]. AB - The Liege University Department of General Practice and the Association of the General Practitioners of Sprimont (Belgian municipality) have jointly drawn up a study protocol named "Evaluation of anti-tetanus cover in rural society". The aim of this epidemiological search program was to answer the 2 following questions: What is the status of anti-tetanus cover among the rural population, according to age, sex and social classes? Is vaccine protection influenced by leisure activities (sport practice, gardening and animal farming at a small scale)? We received 790 answers out of 960 forms sent in a at random selected population (82.3%). A sampling classification has been done by the general practitioners, who have determined 3 categories of patients: unprotected patients (no vaccination for the last 10 years), patients in the process of vaccination, protected patients. Protected patients represent 51.8% of the population. Surprisingly enough, more than 45% of that rural population are not protected. This study clearly shows that two factors highly influence the vaccination cover against tetanus: age (elderly are less protected) and sex (women are always less protected than men). It also emerges from our analysis that the nature of the population's professional activities does not influence the protection rate. In contrast, leisure activities seem to have a favourable impact on the vaccination rate, sport leading to a better protection for both sexes. It is also true in the case of gardening practice, frequent in rural environment. We must then be careful to reach optimal vaccination among a population with an increasing life expectancy, therefore with more and more elderly people, and we must be careful with vaccination among females, presently less protected. The role of the family doctor is paramount to carry out that mission. PMID- 11942186 TI - Rural America: a call for nurses to address mental health issues. PMID- 11942187 TI - Protective and risk behaviors of rural minority adolescent women. AB - Studies of adolescent risk and protective behaviors have focused on identifying modifiable psychosocial variables that predict differential outcomes for subsequent intervention efforts. Research has been scarce in studies of rural minority adolescent women. This study examined the protective and risk behaviors of these women and their relationship to physical or sexual abuse. Rural minority adolescent women endured high levels of psychological distress and many high risk behaviors, yet experienced many protective behaviors. Barriers to health care included access and confidentiality. Physically or sexually abused adolescents endured relatively greater risk and fewer protective behaviors than nonabused adolescents. The prevalence of risk behaviors and abuse among rural minority adolescent women presents a need for development of psychotherapeutic interventions as part of behavioral interventions for risk reduction. These women may benefit from confidential identification and assessment of abuse history and risk and protective behaviors so that appropriate psychological treatment can accompany accessible medical treatment. PMID- 11942188 TI - Complex physical and mental health needs of rural incarcerated women. AB - Despite the growing numbers of incarcerated women, there still remain few systematic analyses of their unique physical and mental health needs. A descriptive design was used in a convenience sample of 30 incarcerated female offenders in a rural detention center to investigate the complex health care needs of this population and formulate appropriate community-based nursing interventions. The participants received a detailed physical and mental health assessment, as well as screening for alcohol and drug abuse. Sixty-three percent of the women reported drug problems and 80% reported alcohol problems, while 84% reported physical or sexual abuse. Serious health problems were identified, including AIDS, STDs, and delirium tremens. Scores on the Global Severity Index of the Brief Symptom Inventory showed that 70% percent of the women were in the clinical range for mental health problems. The scores on the Multidimensional Scale of Perceived Social Support were negatively correlated with the Global Severity Index on the BSI (r = -.377, p = .04) which may indicate some protective effect of social support with regards to psychiatric distress. The data demonstrate a need to develop a brief objective mental health screening test for this specific population to identify psychiatric problems that require immediate attention during the incarceration period. The results also highlight incarceration as an excellent opportunity for the advanced practice nurse to initiate focused health care interventions and other strategies which can foster incarcerated women's re-entry into the community. More research is needed in this underserved group, particularly concerning issues of maternal incarceration, STD prevention, detoxification, psychiatric treatment, and sources of social support. PMID- 11942189 TI - Behind the scenes: spousal coping following permanently disabling injury of farmers. AB - Hidden within the pastoral setting of rural America are the struggles of farm families who must cope with a decreasing economy, increasing technology, and one of the highest injury rates of any industry. Wives of farmers often juggle household responsibilities, hold an off-farm job, and work on the farm. Too often, wives are first on the scene of a serious injury to their spouses. All attention is focused on the farmer but the wife must cope with caregiving, her usual work, and the added full responsibility of the farm. This grounded theory study examined the process of spousal coping after a permanently disabling injury occurred to their farmer husbands. Interviews conducted with 12 farm wives elicited four themes in the coping process. These findings may help identify critical points of mental stress and form interventions to assist spouses of injured farmers in their adjustment to changing roles and responsibilities. PMID- 11942190 TI - The importance of social support for rural mental health. AB - While the mentally ill are considered to be vulnerable as a group, those who are mentally ill and reside in rural communities are at a greater disadvantage due to limited access to health care, a scarcity of resources, and traditional cultural belief systems. Social support has been found to have a direct effect on the well being of families and individuals. Specific strategies are presented which will assist rural health nurses in providing the support systems necessary to help rural residents in managing and coping with stress and mental health problems. PMID- 11942191 TI - Weaving a new safety net of mental health care in rural America: a model of integrated practice. AB - Life in rural America is often idealized, yet rural Americans suffer from mental illness in rates comparable to urban America and require similar types of support and services. However, millions of individuals living in rural areas go without needed mental health services. The dominant care model allows the treatment of mental illness to be delivered by non-mental health professionals with little or no education or training in psychiatric care and who have little desire to provide this type of care, resulting most often in ineffective or inappropriate treatment. Lacking access to appropriate and effective care, rural mentally ill individuals are more often symptomatic than their urban counterparts and may never find relief from the disabling symptoms of treatable mental illnesses. This article will focus on the current state of psychiatric-mental health care in the context of these realities and discuss the impact of the current trend of mental illness being treated by non-mental health professionals. The article will conclude by proposing a model of advanced practice nursing that the authors believe will increase both access and efficacy of treatment for the mentally ill living in rural America. This Integrated Model views the current system of care that completely separates location for traditional physical and mental health care as antithetical to integration and to holism and presents a new model for understanding and provided integrated health care to meet the needs of rural mentally ill individuals and families. PMID- 11942192 TI - The use of reminiscence therapy for the treatment of depression in rural-dwelling older adults. AB - This article explores the potential for the use of reminiscence therapy as an effective means of reducing depression among institutionalized, rural-dwelling elders, especially elderly women. Reminiscence therapy is a nurse-initiated intervention that has the advantages of being cost-effective, therapeutic, social, and recreational for the institutionalized older adult. As a communicative psychosocial process, reminiscence therapy has proven to be a valuable intervention for the depressed elderly client (Cully, LaVoie, & Gfeller, 2001; Haight & Hendrix, 1998; Haight, Michel, & Hendrix, 1998, 2000). It has been shown that depressed elders living in rural areas resist treatment from mental health services for a variety of different reasons (Molinari, Boeve, Kunik, & Snow-Turek, 1999; Neese, Abraham, & Buckwalter, 1999). For those elders, reminiscence therapy may prove an extremely beneficial alternative to more traditional treatment modalities in reducing the effects of depression and depressive symptoms. PMID- 11942193 TI - Factors affecting use of the mental health system by rural children. AB - As many as 20% of children between the ages of 0 and 18 meet the criteria for one or more mental disorders at some point in their lives, with about one half of these being described as being seriously disturbed. Only about one-third of these children and adolescents receive help from the mental health system. Negative outcome expectations toward the mental health system can prevent use of services. This study examined rural parents' expectations about outcomes related to mental health treatment, the provider-client-parent relationships, social and cultural factors, and accessibility to mental health services. The parents' knowledge of the prevalence of mental health disorders in children and adolescents was also examined. Bandura's Social Cognitive Theory served as the conceptual framework for this study. Stigma toward the use of the mental health system was evident. More than half the parents were concerned that mental health professionals would not care for their child. Although negative relationship outcome expectations were revealed, positive treatment outcome expectations also emerged. Structural outcome expectations were not shown to be a major deterrent in receiving care. The belief and hope is that positive outcome expectations toward the mental health system will encourage use of services. PMID- 11942194 TI - Rural woman abuse and sexually transmitted disease: an ethical analysis of clinical dilemmas. AB - Women who have experienced abuse are often offered individual and group psychotherapeutic interventions to help them overcome psychological aftereffects of abuse and avoid future abusive relationships. Clinical cases that are complicated by the existence of abuse and sexually transmitted diseases (STDs) often involve ethical issues concerning patient privacy and legitimate breaches of confidentiality that can potentially harm the individual. There is little known about the experiences of abused rural women with STDs, underscoring the need for modification of existing interventions for this patient population. This paper presents the application of an ethical framework to an actual case involving an abused rural woman with an STD that utilizes the concepts of casuiStry, or case-based reasoning. A methodological tool for ethical analysis of the clinical dilemmas involving evidence-based psychotherapeutic interventions for abused rural women with STDs is used to apply the concepts of casuistry to the development of the taxonomy of cases for clinical practice. PMID- 11942195 TI - [World Medical Association (AMM). Helsinki Declaration. Ethical principles for medical research involving human subjects]. AB - The role of nurses in the Clinical Ethic Committees (ECs) was discussed in a seminar organised by Assistenza Infermieristica e Ricerca and the Mario Negri Research Institute. A recent Italian law of 1998 regulates the functioning and the composition of the local ECs, where at least a nurse must be present. The actual and potential role of the nurses has been discussed from different points of view with and by other professions represented in ECs. Few model experiences were presented by nurses member of ECs. Based on these contributions an important discussion developed, which led to the identification of the main areas of interest and of the key priorities for the development of a more active involvement of nurses in issues specifically related to informed consent, prospective monitoring of the trial implementation, continuity of care across experimental protocols and routine practices. The main contributions to the seminar are reported, together with proposal for developing a specific nursing culture and practice to ECs and clinical trials. PMID- 11942196 TI - [Questions and answers on the patient with bladder catheterization]. PMID- 11942197 TI - [Millions of refugees: a burden for the South of the world]. PMID- 11942199 TI - [Shortage in nursing]. PMID- 11942198 TI - [The 5 pitfalls of education]. PMID- 11942200 TI - [Management of turnover of nursing personnel: descriptive study]. AB - The strategies of management of the turnover of nursing personnel have been explored in 5 hospitals of the Veneto region. A questionnaire distributed to 200 permanent staff nurses moved for different reasons to another ward during the year 1998, was returned by up to the 62.5% of the recipients. Sixty nurses changed the ward at their request, 18 had to move only for work reasons and 47 for a combination of both reasons. The main reasons (50%) for moving nurses for work reasons (18/125) is the lack of personnel or the restructuration or closure of wards with reduction of the number of beds (50% of the moves). Professional development and acquisition of new competences and skills is the main reason (42%) for nurses who ask to change ward at their request. The reasons for changing the ward are obscure for 1/3 of nurses moved for work reasons. Overall mobility is valued as opportunity for professional development, but it is suggested it should be mainly at personnel's request and after an evaluation of the curriculum. PMID- 11942201 TI - [Variability in the management of patients with bladder catheterization in the ULSS 18 of Rovigo]. AB - A survey on the practices related to the management of patients with indwelling urinary catheter was organised in two hospitals and health districts, with the aim of identifying problems or incorrect habits. Thirty-five nurses were interviewed with a questionnaire exploring different practices and procedures related to the catheter care. While practices were overall satisfactory and homogeneous across wards, few problems were identified: the ill defined practice of bladder training before catheter removal; the difficulties in maintaining a closed system; an improper use of sterile sacs. The data were presented and discussed during two meetings attended by 186 nurses where the emerging problems and the practicability of changes in habits and ward protocols were thoroughly discussed. PMID- 11942202 TI - [Nurses in the ethics committees for clinical experimentation]. PMID- 11942203 TI - [Nurses in ethical committees for clinical experimentation. Guidelines]. PMID- 11942204 TI - [Role of nurses in ethical committees: an experience]. PMID- 11942205 TI - [Informed consent in clinical experimentation: typology of the errors most frequently found by the committee for experimentation of the Lombardy region]. PMID- 11942207 TI - [Management and nursing practice (interview by Harald Verworner)]. PMID- 11942206 TI - [Role of nurses in ethical committees. Status and projects]. PMID- 11942208 TI - [Personnel planning for the general nursing unit]. PMID- 11942209 TI - [Nursing management]. PMID- 11942210 TI - [Hospital management and theoretical principles]. PMID- 11942211 TI - [Didactic of nursing diagnosis at the General Health and Nursing School of Krems on the Donau A.O. Hopsital]. PMID- 11942212 TI - [Conversion of nursing diagnosis as an integrated element of the nursing process. II]. PMID- 11942213 TI - [Nursing diagnosis]. PMID- 11942214 TI - [Nursing diagnosis--basic principles and practice references. Or: "Guidelines for failure in implementing nursing diagnoses"--by Manfred Zottl, DG KP, nursing diagnosis network]. PMID- 11942215 TI - [Welcome to the 3rd century]. PMID- 11942216 TI - [Professional pictures: a collage. Honored, controlled, humbled]. PMID- 11942217 TI - [Society and nursing care. The human dimension is at the center]. PMID- 11942218 TI - [We must become louder (interview by Urs Luthi)]. PMID- 11942219 TI - [Laptop as defending shield]. PMID- 11942220 TI - [Nursing care on the floor. Even miracles begin on the floor]. PMID- 11942221 TI - [HIV. Despite therapy--family is silent]. PMID- 11942222 TI - [Steps, direction, independence]. PMID- 11942223 TI - [Benchmarking for tomorrow]. PMID- 11942224 TI - [Free and responsible: can we become that?]. PMID- 11942225 TI - [Atlas]. PMID- 11942226 TI - [Ephemeral traces. Reflexions on human reality]. PMID- 11942227 TI - [What family dimension for tomorrow?]. PMID- 11942229 TI - [What name does the future have now?]. PMID- 11942228 TI - [What will happen in 2000?!]. PMID- 11942230 TI - [BSE--bovine spongiform encephalopathy & Creutzfeldt-Jakob disease]. PMID- 11942231 TI - [Improvement projects in public health. Many good ideas are born in general practice for management of patients and citizens, but only a few are systematically implemented, evaluated and made public. Why is that so?]. PMID- 11942234 TI - [ICN Congress. What is the value of being a member or an international organization?]. PMID- 11942235 TI - [Nursing diagnosis--basic principles & general practice references. Or: Guidelines on failures in implementing nursing diagnoses. II]. PMID- 11942236 TI - [International education--Eu program. AZW Innsbruck Health and Nursing School participates in the EU program of the Leonard Da Vinci project]. PMID- 11942237 TI - Franchising. Where's the beef? AB - Franchising is most appropriate to services which can be simply defined. Introducing it to the NHS will raise issues about accountability. Those taking on franchises may feel they need more freedom than is currently available. The scope for franchising in the NHS seems limited. PMID- 11942238 TI - NHS memories. 1984 and all that ... an editor remembers. PMID- 11942239 TI - Refugee health. Speak easy. AB - Many NHS staff are unsure about the rights of asylum seekers and refugees to healthcare. Asylum seekers and refugees have little knowledge of the NHS. Workshops including refugee speakers helped dispel misconceptions and were highly rated by primary care staff. The format is very transferable. PMID- 11942240 TI - E-novation. Another write-off. PMID- 11942241 TI - E-novation. The next big thing. War and peace. PMID- 11942242 TI - E-novation. The Metaman prophecies. PMID- 11942243 TI - E-novation. All within reach. PMID- 11942244 TI - E-novation. Cradle to grave. PMID- 11942245 TI - Further examining BMS (Building Management Systems) issues. PMID- 11942246 TI - Laundering advances. PMID- 11942247 TI - Decontamination upgrading progresses. AB - Standards of decontamination of surgical instruments in the NHS in England have come under close scrutiny, and some significant improvements have been achieved. Action plans, developed to ensure the rate of improvement is maintained, are described in this article which is derived from the Department of Health/NHS Estate document 'A review of the decontamination of surgical instruments in the NHS in England'. The Institute of Healthcare Engineering and Estate Management is one of the organisations involved in the programme of work on decontamination. PMID- 11942248 TI - Renovate or replace? AB - The Cross Cancer Institute completed final facility is locked in on all sides with minimal room for future growth. Would a new facility in a new location have been a better option? From an engineering perspective, it would have been less complicated and frustrating to build new with a new stand-alone plant sized for future expansion capability. From a medical/research perspective, the location adjacent to the university was and still remains invaluable. PMID- 11942249 TI - Utilising wireless information transfer. PMID- 11942250 TI - On alert for violence. PMID- 11942251 TI - Holistic approach delivers first rate results. PMID- 11942253 TI - New IC/HD unit at Bedford. PMID- 11942258 TI - Groom for more smart technology. PMID- 11942259 TI - Evidence-based management: using serial firm trials to improve diabetes care quality. AB - BACKGROUND: The design of delivery systems that can truly conduct continuous quality improvement (CQI) as a routine part of clinical care provision remains a vexing problem. The effectiveness of the "computerized firm system" approach to chronic disease CQI was examined, with diabetes as the focus of a 5-year case study. METHODS: A large family medical center had been divided into two parallel group practices for reasons of efficiency. These frontline structures (also known as primary care "firms") were supported to serially adapt and evaluate selected CQI interventions by first introducing process changes on one firm but not the other and comparing the groups. Because all the required longitudinal data were contained in a computerized repository, it was possible to conduct these controlled "firm trials" in a matter of months at low cost. RESULTS: During a 3 year period, implementation of point-of-service reminders and a pharmacist out reach program increased recommended glycohemoglobin (HbA1c) testing by 50% (p = 0.02) and reduced the number of diabetic patients inadequately controlled by 43% (p < 0.01). Following this outcome improvement, patients exhibited a 16% reduction in ambulatory visit rates (p = 0.04). The observed outcome improvement, however, was reversed during the subsequent 2 years, when staffing austerities forced by unrelated declines in clinic revenue caused the withdrawal of trial interventions. CONCLUSIONS: The processes and outcomes of diabetes care were improved, demonstrating that CQI and controlled trials are not mutually exclusive in moving toward the practice of evidence-based management. Health care systems can, by conducting serial firm trials, become learning organizations. CQI programs of all kinds will likely never flourish, however, until quality improvement and reimbursement mechanisms have become better aligned. PMID- 11942260 TI - Assessing clinical pathways use in a community hospital: it depends on what "use" means. AB - BACKGROUND: Many benefits have been associated with the use of clinical pathways, yet developing them can be costly, and implementing them is not always successful. A 300-bed Midwestern community hospital began a clinical pathways program in 1995, and by fall 1998, 15 pathways were in various stages of implementation, with 3 under development. Many challenges had been encountered, but hospital leaders were eager to find ways to increase pathway use. METHODS: A qualitative case study design was used to investigate four clinical pathways, two perceived as being "used" and two that were perceived as "not used". Each pathway was analyzed as a separate case, followed by cross-case analysis. Qualitative data were collected in 65 semistructured interviews with administrators, physicians, physicians' office staff, nurses, and allied health professionals at the hospital. Data were also collected through observation and document analysis. RESULTS: The two used pathways had been introduced as part of a larger change in care, whereas the two pathways not used had been introduced as stand-alone innovations. Confusing and inadequately developed aspects of the hospital's clinical pathways program included its purposes, the definition of pathway use, pathway procedures, accountability, education, and incentives. A new case management department, ongoing administrative support, and a sophisticated medical information system were viewed as supports for continued growth in the program. CONCLUSIONS: Implementation of clinical pathways was delayed and complicated by the varied perceptions of the program among stakeholders. Lack of clarity and consistency in how information about the program was communicated made it difficult for clinicians to develop a shared understanding of clinical pathways. PMID- 11942261 TI - Developing a quality measurement tool and reporting format for long- term care. AB - BACKGROUND: Quality measurement in long term care (LTC) presents many challenges: the lack of a uniform definition of quality, the existence of multiple domains for measurement, a multitude of potential perspectives, and regulatory influences that emphasize measurement only of poor quality. Research efforts have yet to solve the issues of measurement; however, operators of long term care facilities must use the current state of the art in quality measurement as the basis for their quality improvement efforts. A project was commissioned by management of a large integrated delivery system with a robust network of LTC facilities who wished to implement a continuous quality improvement process on the basis of a measurement tool that provides a comprehensive resident-centered assessment of quality. The objectives of this project, therefore, were to identify domains of quality, to select and adapt validated instruments for measurement within each domain, to pilot test a data collection process, and to develop an operational quality profiling report format for LTC facilities. DESIGN AND METHODS: Using an expert panel and the LTC research literature, an operational measurement tool was developed, consisting of four domains of quality: organizational, clinical, environmental, and social. DISCUSSION: A pilot study conducted in two nursing facilities demonstrated that the data collection process could be operationalized within tight resource and budgetary constraints. The development of an operational quality assessment tool enables management to take a consistent view of diverse institutions, focusing in detail on quality of care as it is perceived by residents. The tool allows evaluation of trends over time and comparison to external norms. PMID- 11942262 TI - Reporting outcomes and other issues in patient safety: an interview with Albert Wu. Interview by Steven Berman. PMID- 11942263 TI - [Year 4 with the new health and nursing care regulation. From helping profession to helping the profession]. PMID- 11942264 TI - ["The University lives"! The health and nursing care regulation in it's 4th year- the IDS nursing since in it's 3rd year by Mag. Elisabeth Rappold and Paul Resetarics, IDS Nursing Science Tutorial]. PMID- 11942265 TI - [Education: experiences and perception of the 179th regulation of the Federal Minister for Occupation, Health and Social Issues in the elevated service of health- and nursing care. BGBI. II. 179/1999 from the viewpoint of an educator]. PMID- 11942266 TI - [Experience of supervised practice by educators for health- and nursing care in nursing education]. PMID- 11942267 TI - [1997-2001: 4 years health and nursing care regulation. A significant step for home care nursing service--the first time that the extramural service is specifically mentioned in the regulation]. PMID- 11942268 TI - ["Nursing research: current developmental trends and challenges"]. PMID- 11942269 TI - [Public health and nursing]. PMID- 11942270 TI - [Austria's role in international organizations is traditional. But how are details resolved for joining projects involving international aid?]. PMID- 11942271 TI - [Tserona City]. PMID- 11942272 TI - ["So rich and yet so poor!"]. PMID- 11942273 TI - [Tuberculosis--a problem at the threshold of the 21st century. A report on the current status with the example of Vienna by Christian Schotta, DGKP--Otto Wagner Hospital]. PMID- 11942274 TI - Follow-up of renal function in treated and untreated older patients with isolated systolic hypertension. PMID- 11942275 TI - [Response to the comment by K.-J. Winzer and J. M. Muller. Nonpalpable breast carcinomas--vacuum biopsy confirmation and surgical management]. PMID- 11942277 TI - Prison health care: opening the debate. PMID- 11942276 TI - Nurses are overloaded with too many policies. PMID- 11942278 TI - Prison health care: opening the debate. PMID- 11942279 TI - Personal care is part of holistic nursing. PMID- 11942280 TI - Pain assessment and treatment in the managed care environment. A position statement from the American Pain Society. PMID- 11942281 TI - We must never forget what nursing is about. PMID- 11942282 TI - We must never forget what nursing is about. PMID- 11942283 TI - We must never forget what nursing is about. PMID- 11942284 TI - Students are not being supervised in practice. PMID- 11942285 TI - Need for a continuum in prison health care. PMID- 11942286 TI - Research notes. Mentoring. PMID- 11942287 TI - Ethics self-assessment. PMID- 11942288 TI - Reducing your risk for rectal cancer. PMID- 11942289 TI - ICAAC. New resistance findings presented at annual conference. PMID- 11942290 TI - You can work it out. The art of keeping your relationship alive. PMID- 11942292 TI - Taking action with art. PMID- 11942291 TI - 'You have the right to remain safe.' L.A.'s Correct HELP goes behind bars to save lives. PMID- 11942293 TI - Know your rights as a disabled passenger. PMID- 11942294 TI - Breaking the cycle of using. PMID- 11942295 TI - Crystal, I loved you. PMID- 11942296 TI - Bush AIDS policy chief takes firsthand look at L.A.'s HIV community. PMID- 11942297 TI - Surfing for straight positives. PMID- 11942298 TI - Recovering from rape. Making space for healing is critical for survivors of rape. PMID- 11942299 TI - Legislating good sense. It's time for medical nutrition therapy to be part of standard care for people with HIV/AIDS. PMID- 11942300 TI - Accessing ADAP. Enrollment services for the state Drug Assistance Program are now available at AIDS Project Los Angeles. PMID- 11942301 TI - Nursing homes need better regulation. PMID- 11942302 TI - Why hospital league tables? We want action! PMID- 11942303 TI - Patients with dementia deserve to be fed. PMID- 11942304 TI - [Report of study residence at the department of colorectal surgery at the UIC Medical Center and at Mountain Home VA Medical Center of the East Tennessee State University in Johnson City]. PMID- 11942305 TI - [Report on study residence at the department of colorectal surgery and minimal invasive surgery of Mount Sinai Hospital New York]. PMID- 11942307 TI - Are misconduct cases the tip of an iceberg? PMID- 11942306 TI - Teamwork: but how do we actually do it? PMID- 11942308 TI - Agency nurses try hard to do their best. PMID- 11942309 TI - [Current status and future of studies on micordomain(discussion)]. PMID- 11942310 TI - Clinical areas need clinical educators. PMID- 11942311 TI - Relationship-centred care: what's in a name? PMID- 11942312 TI - Variations in the subclavian-axillary arterial system. AB - OBJECTIVE: The objective of this study is to report on the subclavian axillary arterial system variations observed in the dissecting room. METHODS: The morphological study of 106 formalin fixed upper limbs of adult human cadavers of both sexes was carried out at human anatomy laboratory of College of Medicine, King Saud University, Riyadh, Kingdom of Saudi Arabia over a 5 year period, to observe the distribution pattern of the subclavian axillary arterial system. RESULTS: The arterial variations in the upper extremities are quite common and may occur at the level of the thoracic outlet, axilla, arm, cubital fossa and the hand. In the present study, the anomalous topographic pattern of the subclavian axillary arterial system was revealed in 7.5% of the cadavers, whereas 92.5% exhibited classical pattern of the regional arterial anatomy. The study showed: 1. An aberrant right subclavian artery arising from the arch of aorta, distal to the left subclavian artery. 2. A bilateral common subscapular-circumflex humeral trunk (3.8%) emerging from the 3rd part of the axillary artery (branching into the circumflex humeral and thoracodorsal arteries. 3. A bilateral thoracohumeral trunk arising from the 2nd part of the axillary artery (1.9%) and branching into the lateral thoracic, circumflex humeral, subscapular and thoracodorsal arteries. These anomalies were accompanied by anomalous insertion of flexor digitorum superficialis muscle, a bilateral digastric muscle formed by the abductor pollicis longus and brevis, and an anomalous formation of median nerve by 3 roots. CONCLUSION: The aberrant right subclavian artery associated with right non recurrent laryngeal nerve and variant branching pattern of the axillary artery are of interest to anatomists, surgeons and radiologists and suggested that these anomalies must be evaluated pre-operatively. PMID- 11942313 TI - To: Lindner T, Cockburn BN, Bell GI (1999). Molecular genetics of MODY in Germany. Diabetologia 42: 121-123. PMID- 11942315 TI - Complete glucokinase deficiency is not a common cause of permanent neonatal diabetes. PMID- 11942314 TI - To: T.J. Wilkin (2001) The accelerator hypothesis: weight gain as the missing link between Type I and Type II diabetes. Diabetologia 44: 914-921. PMID- 11942316 TI - Losartan attenuates endothelial hyperpermeability responses to VPF and high glucose but not angiotensin II. PMID- 11942318 TI - Professional misconduct case studies. Case 54: getting back on the nursing register. Three cases of nurses wanting to be restored to the register. PMID- 11942317 TI - Flamazine is not a debriding agent. PMID- 11942319 TI - Pressure/cross-sectional area relations in the proximal urethra of healthy males. Part III: the time dependent pressure response following forced dilation: standardization of a technique. AB - The urethral response to a sudden forced dilation was studied in ten healthy male volunteers aged from 27 to 71 years. Measurements were performed from the bladder neck and beyond the region of high pressure using a specially designed probe. The pressure response after dilation showed a characteristic course, which could be described by a double exponential function of the form: P(t) = P(equ) + P(alpha)e(-t/tau(alpha) + P(beta)e(-t/tau(beta) where P(t) is pressure at time t, P(equ) is equilibrium pressure after dilation, P(alpha) and P(beta) are pressure decay, and tau(alpha) and tau(beta) are time constants. The size and velocity of dilation, as well as the degree of distension before dilation, proved of significance for the magnitude of the pressure response. The characteristics of the pressure response are given by the properties of the periluminal structures strained during dilation, and are thus predominantly determined of elastic, collagen, muscular, and glandular components. However, a high degree of relaxation after straining, and a modest stiffness, indicates that the muscular component dominates the response. The significance of the prostatic tissues remains unclear. PMID- 11942320 TI - In vitro comparison of transurethral vaporization of the prostate (TUVP), resection of the prostate (TURP), and vaporization-resection of the prostate (TUVRP). AB - PROBLEM: Transurethral vaporization of the prostate (TUVP) and vaporization resection of the prostate (TUVRP) ("vapor cut, band electrode") seem to be alternatives to conventional resection of the prostate (TURP). For TUVP and TUVRP, little in vitro data has yet been published. The aim of this study was to determine settings for optimal performance with TUVP and TUVRP and to investigate electrosurgical parameters relevant to safety. METHODS: Standardized experiments were performed on porcine muscle. Mass loss and coagulation zones were measured optically. Additionally, electrical parameters were recorded. RESULTS: The maximum tissue ablation rates were 3.8 cm3 per min and 6.1 cm3 per min for TUVP and TUVRP, respectively, compared to 6.5 cm3 per min for TURP. The maximum coagulation depths reached 2.1 mm (TUVP), 1.4 mm (TUVRP), and 0.9 mm (TURP). Optimal in vitro settings for TUVP/TUVRP/TURP were as follows: generator power of 250/120/90 W, drag speed of 5/15/20 mm/s, and pressure of 0.40/0.15/0.05 N. Different power generators and electrodes showed considerably varying performance. The energy to remove 1 g of tissue averaged 7.500 J (TUVP), 620 J (TUVRP). and 400 J (TURP). CONCLUSIONS: These results allow quantification of the influence of different variables on TUVP, TUVRP, and TURP in vitro. The TUVP proves to be an effective ablation alternative. Nevertheless, a 15 to 20 times higher energy demand has to be considered. TUVRP combines excellent ablation features with greater coagulation volumes, indicating better hemostasis. PMID- 11942321 TI - Effects of chronic cadmium exposure on contractility of the rat detrusor. AB - The chronic effects of Cd2+ on the myogenic contractions induced by acetylcholine (ACh), and the neurogenic contractions induced by electrical field stimulation (EFS) of the rat detrusor were investigated. Wistar Kyoto rats weighing 150-250 g were randomly divided into four groups each containing ten animals. Three groups received intraperitoneal Cd2+ (0.25, 0.5 and 1 mg/kg, respectively) dissolved in saline twice a week for 3 months. The control group received only saline (0.3 ml). At the end of 3 months, the urinary bladders were surgically removed and a strip of detrusor was prepared from each bladder. An atomic absorption device and the standard addition method were used to determine blood levels of Cd2+ and the Cd2+ levels of the remaining parts of each bladder. The responses of the detrusor strips were studied in organ chambers. The tissues were first treated with ACh and then with EFS. The responses were recorded by isotonic transducers. The tissue Cd2+ levels were significantly increased in the Cd2+ treated rats in a dose-dependent manner except in the 0.25 mg/kg Cd2+ treated group. ACh-induced contractions were significantly attenuated only in the 1 mg/ kg Cd2+ treated rats. The contractions induced by EFS were significantly decreased in all of the Cd2+-treated groups. but there were no significant differences between the groups. This study showed that Cd2+ exposure for 3 months impairs neurogenic and myogenic contractile activity in the rat detrusor muscle. This action seems to be at least partly due to an inhibition of the cholinergic muscarinic system. This may have clinical implications for people who are exposed to Cd2+. PMID- 11942322 TI - Effects of estrogen on the relaxation response of rabbit clitoral cavernous smooth muscles. AB - We investigated whether estrogen has a peripheral effect on the smooth muscle relaxation response of the clitoris, similar to that of androgens in cavernous tissues of the penis. Forty-two New Zealand White female rabbits (2.3-2.7 kg) were randomly divided into control (sham operation) and bilateral oophorectomy groups (acute and chronic groups with or without estradiol replacement). The acute and chronic groups were killed at 5 days and 12 weeks post-oophorectomy, respectively. Relaxation responses of the clitoral cavernous strips to endothelium-dependent and independent vasodilators and electrical field stimulation (EFS) were observed. Serum total cholesterol and low-density lipoprotein increased significantly and acetylcholine-induced relaxation of the clitoral strips wa significantly attenuated in the oophorectomy group with chronic estrogen deficiency but not in the chronic oophorectomy group with estradiol replacement or in the acute estrogen deficiency group. No significant differences in the relaxation responses of the cavernous strips to sodium nitroprusside or to the non-adrenergic, non-cholinergic-selective relaxation response to the EFS were found among the control, acute and chronic estrogen deficiency groups. These findings suggest that estrogen may not have a direct peripheral effect on clitoral cavernous smooth muscle relaxation. Hypercholesterolemia might be a possible explanation for the impaired endothelium dependent relaxation of clitoral smooth muscle in chronic estrogen deficiency. PMID- 11942324 TI - Calcium oxalate stone disease: role of lipid peroxidation and antioxidants. AB - Membrane injury facilitated the fixation of calcium oxalate crystals and subsequent growth into kidney stones. Oxalate-induced membrane injury was mediated by lipid peroxidation reaction through the generation of oxygen free radicals. In urolithic rat kidney or oxalate exposed cultured cells, both superoxide anion and hydroxyl radicals were generated in excess, causing cellular injury. In hyperoxaluric rat kidney, both superoxide and H2O2-generating enzymes such as glycolic acid oxidase (GAO) and xanthine oxidase (XO) were increased, and hydroxyl radical and transition metal ions, iron, and copper were accumulated. The lipid peroxidation products, thiobarbituric acid-reactive substances (TBARS), hydroperoxides, and diene conjugates were excessively released in tissues of urolithic rats and in plasma of rats as well as stone patients. The accumulation of these products was concomitant with the decrease in the antioxidant enzymes, superoxide dismutase (SOD), catalase, glutathione peroxidase (GPx), and glucose-6 phosphate dehydrogenase (G6PD) as well as radical scavengers, vitamin E, ascorbic acid, reduced glutathione (GSH), and protein thiol. All the above parameters were decreased in urolithic condition, irrespective of the agents used for the induction of urolithiasis. Oxalate binding activity and calcium oxalate crystal deposition were markedly pronounced, along with decreased adenosine triphosphatase (ATPase) activity. Lipid peroxidation positively correlated with cellular oxalate, oxalate binding, gamma-glutamyl carboxylase, and calcium level and negatively correlated with GSH, vitamin E. ascorbic acid, and total protein thiol. Antioxidant therapy to urolithic rats with vitamin E, glutathione monoester, methionine, lipoic acid, or fish oil normalised the cellular antioxidant system, enzymes and scavengers, and interrupted membrane lipid and protein peroxidation reaction, ATPase inactivation, and its associated calcium accumulation. Antioxidant therapy prevented calcium oxalate precipitation in the rat kidney and reduced oxalate excretion in stone patients. Similarly, calcium oxalate crystal deposition in vitro to urothelium was prevented by free radical scavengers such as phytic acid and mannitol by protecting the membrane from free radical-mediated damage. All these observations were suggestive of the active involvement of free radical-mediated lipid peroxidation-induced membrane damage in the pathogenesis of calcium oxalate crystal deposition and retention. PMID- 11942323 TI - Selenium reduces high energy shock wave-induced renal injury in rats. AB - Using an in vitro model with Madin-Darby canine kidney (MDCK) cells, we showed that shock wave-induced renal injury could be ameliorated by selenium. We examined the influence of selenium, a free radical scavenger, in shock wave induced tubular cell injury in vivo. Male rats were randomly assigned to three groups: 1 control (n= 18), 2 selenium (n = 18), 3 sham treatment (n = 4). Groups 1 and 2 were treated with 500 shock waves on each kidney. Animals assigned to group 3 (sham treatment) received only anesthetics. Selenium (80 microg/kg per 24 h intraperitoneally) was given to the animals in group 2 for 5 days, starting 1 day before shock wave exposure. Urine was collected for 8 h on the day before and immediately, 1, 7 and 28 days after shock wave exposure (SWE) for the measurement of urine volume, N-acetyl-beta-glucosaminidase (NAG), beta-2-microglobulin (beta2 M), and creatinine. Blood was taken from these rats on day 1 after SWE for the determination of creatinine and the calculation of the creatinine clearance (CCr). After SWE, there was a significantly increased diuresis in group 1 and 2. The excretion of NAG and beta2 M was also increased in both groups. These changes were significantly less pronounced in the selenium treated rats. CCr was higher in the selenium group than in the controls. No changes were observed in the sham treated group. These results demonstrate that selenium is able to ameliorate the damaging effects of high energy shock waves on renal tissue not only in vitro, but also in vivo. PMID- 11942325 TI - The role of superoxide anion in the regulation of epidermal growth factor or the expression and proliferation of its receptor in prostate cancer cell line PC3. AB - The purpose of this study was to investigate the role of superoxide anion(O2-) in the regulation of epidermal growth factor (EGF) or epidermal growth factor receptor (EGFR) expression and proliferation in the prostate cancer cell line PC3. Cell proliferation was tested by a 3-(4,5-dimethylthiazol-2-yl) diphenyltetrazolium bromide (MTT) assay in the presence of O2-, EGF or their combination. Immunohistochemistry was carried out to assay the expression of EGF or EGFR. EGF or EGFR mRNA expression in the cells treated with O2- was examined by in situ hybridisation. The proliferation was significantly inhibited by O2- in a concentration-dependent manner ranging from 9 to 36 micromol/l nicotinamide adenine dinucleotide (NADH) combined with 2-8 micromol/l N-methylphenazonium methyl sulfate (PMS). The enhancement of proliferation induced by 5 ng/ml EGF was significantly overcome by O2-. Although O2- was not able to alter EGFR mRNA expression, O2- at the concentration of 18 micromol/l NADH and 4 micromol/l PMS reduced EGFR protein expression. O2- at the concentration of 18 micromol/l NADH and 4 micromol/l PMS can downregulate EGF and EGF mRNA expression. PMID- 11942326 TI - N-acetyl-L-cysteine enhances chemotherapeutic effect on prostate cancer cells. AB - Transcription factor nuclear factor kappaB (NF-kappaB) controls gene expression of a number of genes, including cytokines such as interleukin-6 (IL-6), granulocyte-macrophage (GM)-CSF, and interleukin-8 (IL-8). IL-6 is known to play important roles in the growth of prostate cancer cells, activation of androgen receptor, and prostate-specific protein expression. NF-kappaB is activated by extracellular signals such as proinflammatory cytokines, chemotherapeutic reagents, and radiation. Here we demonstrate that cisplatin (CDDP) and etoposide (VP-16) induce nuclear translocation of NF-kappaB in prostate cancer cell lines, followed by secretion of IL-6. We also demonstrated that the growth of hormone independent prostate cancer cell lines can be inhibited by the anti-NF-kappaB reagent N-acetyl-L-cysteine (NAC). These observations indicate that NF-kappaB can be a target of new adjuvant therapy against hormone refractory prostate cancer. PMID- 11942327 TI - The effects of meglumine gamma linolenic acid (MeGLA) on an organ culture model of superficial bladder cancer. AB - The objective of this research was to assess the effects of the novel intravesical drug MeGLA in a physiologically representative model of superficial bladder cancer. Petri dishes were used to culture 5 mm square explants of rat bladder in minimal volumes of supplemented culture medium. Parental and resistant MGH-U1 urothelial cancer cells were transfected with a green fluorescent protein (GFP) vector. Transfectants were purified by flow cytometry. Cells were seeded onto the prepared organ cultures and imaging was performed using confocal microscopy. Confirmation of the tumour colonies was done using scanning electron microscopy. MeGLA was added at various concentrations to the colonies and its effects noted over several days. Results showed that colonies of GFP-MGH-U1 cells established themselves on the explants and could be identified by confocal microscopy. The colonies could then be followed over several days. The colonies were able to survive high concentrations of the drug of up to 1 mg/ml, 400 times the IC > 90% for monolayers and equivalent to doses in clinical use. We conclude that MeGLA is less effective in this system than on monolayer cell lines. However, it showed cytotoxic effects which were comparable to those seen with conventional agents in the same system. PMID- 11942328 TI - Histogenesis of nonurothelial carcinomas of the urinary bladder from pre-existent transitional cell carcinomas. A histopathological and immunohistochemical study. AB - The histogenesis of nonurothelial carcinomas of the urinary bladder is difficult to understand, since the bladder is normally lined exclusively by transitional cell epithelium. To gain more insights into the pathogenesis of nonurothelial carcinomas, the morphology and immunohistochemistry of transitional cell carcinomas (TCC), mixed transitional cell and nonurothelial carcinomas, and pure nonurothelial carcinomas were comparatively studied. Of papillary and of nonpapillary (solid) TCC (overall incidence 6.8%), 4.8% and 15.4%, respectively, disclosed foci of altered celllular and architectural phenotypes, consisting of squamous epithelium, pseudoglandular formations, and true glands with or without mucus production. The diverse phenotypic variants develop obviously by a metaplastic process as a result of the well-known inherent potential of the urothelium to undergo several pathways of cellular differentiation. There is strong evidence that squamous cell carcinomas arise secondarily from a squamous metaplasia and adenocarcinomas from metaplastic glandular epithelium within pre existing TCC following complete carcinogenic transformation of the initially bland-looking metaplastic tumor cells. The metaplastic origin of nonurothelial bladder carcinomas is supported by immunohistochemical findings. The high molecular weight cytokeratin 34betaE12 identifies tumor cells with squamous characteristics, helping to explain the development of squamous cell carcinomas. Secretion of MUC5AC apomucin is assumed to play a central role in the histogenesis of nonurachal mucus-producing adenocarcinomas, including signet ring cell carcinomas. Metaplastic phenotypic variants of TCC should be recognized as distinct tumor entities with the potential to transform into nonurothelial carcinomas and thus possibly implying a poorer clinical outcome than typical, uniform TCC. PMID- 11942329 TI - Decreased urinary bladder apoptosis in a rabbit model of diabetes mellitus. AB - Urinary bladder dysfunction is a recognised complication of diabetes mellitus (DM). This is thought to be partly related to altered bladder morphology as DM is associated with increased bladder weight. In DM, increased cellular proliferation is well established. However, there is evidence that in other pathological states affecting the urinary tract, altered apoptosis may also play a role. We therefore used a rabbit model to investigate whether there are any changes in bladder apoptosis with DM. Diabetes was induced in adult New Zealand white rabbits. Age matched controls were also used. After 6 months, the bladders were excised and weighed. The TUNEL technique was used to detect and quantify apoptosis in both DM and age-matched control bladders. Diabetes was confirmed as this group had significantly (P<0.001) elevated serum glucose-compared to controls. The bladder weights were also significantly (P<0.001) greater in the DM rabbits. Apoptosis was significantly (P<0.001) decreased in the urothelial cells of the DM bladders. Our results confirm previous findings that DM is associated with increased bladder weight. Although this is associated with increased cellular proliferation, we have demonstrated that decreased apoptosis may also play an important role. Therefore, decreased apoptosis may be important in the pathophysiology of DM cystopathy. PMID- 11942330 TI - Pressure/cross-sectional area relations in the proximal urethra of healthy males: the time dependent pressure response following forced dilation. Part IV: results in healthy volunteers. AB - The significance of the anatomical location and age on the urethral response to a sudden forced dilation was studied in 30 healthy males aged 23-85 years. The pressure decay after dilation was fitted with a double exponential function of the form: P(t) = P(equ) + P(alpha)e(-t/tau(alpha) + P(beta)e(t/tau(beta) where P(t) is pressure at time t, P(equ) is equilibrium pressure after dilation, P(alpha) and P(beta) are pressure decay, and tau(alpha) and tau(beta) are time constants. The pressure response was highly affected by the location of the measurement, with the maximum values of the pressure components in the high pressure zone and significantly lower values in the prostatic part of the urethra. The variation in pressure thus concurs closely with the density of the striated rhabdosphincter. No significant correlation between age and the pressure components could be demonstrated, whereas the velocity of the pressure decay following dilation proved significantly related to age in all urethral segments. The causal background for this correlation is uncertain, but may be discovered in age dependent changes in the periluminal tissue composition, or in changing neuromuscular activity in these structures. PMID- 11942331 TI - Nurse crisis is caused by more than shortages. PMID- 11942332 TI - Nurse crisis is caused by more than shortages. PMID- 11942333 TI - Nurse crisis is caused by more than shortages. PMID- 11942334 TI - Passiveness will not advance nursing. PMID- 11942335 TI - Wrapped in silence: psychotherapists and confidentiality in the courtroom. AB - The impact of the Austrian Psychotherapy Act, which, in contrast to legal provisions in the United States, does not provide for any exceptions to breach confidentiality, is compared with the effects of U.S. law on dealing with confidentiality. The authors investigated the impact of this law in light of three common situations in psychotherapy that may jeopardize strict confidentiality: treating potentially dangerous patients, giving testimony, and serving as a psychotherapist in prison. Under the strict provisions of the Austrian Psychotherapy Act, a breach may be excusable in the case of a highly probable danger, but Austrian psychotherapists cannot be obliged to serve as witnesses or as experts in civil or criminal cases, as American psychotherapists can. Psychotherapy in prison, where release is contingent on the success of the therapy and the divulging of information could be in the interests of the patient as well as the court and the public, requires a modified dealing with confidentiality. PMID- 11942336 TI - Examining the foreseeable: assisted suicide as a herald of changing moralities. AB - After her intense battle for the decriminalization of assisted suicide in the Supreme Court of Canada, Sue Rodriguez committed suicide with medical assistance in 1994. Following her suicide, government and law representatives remained silent and no criminal charges were ever brought against the person(s) who presumably assisted Ms Rodriguez in her death. This apparent non-intervention of criminal law is examined in view of the useful role that the Rodriguez event may have played in a possible shift in the dominant morality. It is argued that the Rodriguez assisted suicide may have been a useful 'crime' (in the Durkheimian sense) in that it brought to the fore the possibility that social conditions- which made the 'crime' possible--may no longer be in harmony with conventional morality. Similarly to Socrates' crime, the Rodriguez case can be seen as an anticipation of a new morality. It can be analyzed as a prelude to alterations, as directly preparing the way for changes in the dominant morality. The role of criminal law as a preferred mode of moral regulation is also examined in relation to the moral demands and expectations that arose during as well as after the judicial saga. PMID- 11942337 TI - What is an employee benefit plan?: ERISA preemption of "any willing provider" laws after Pegram. AB - This note considers the implications of a recent Supreme Court decision, Pegram v. Herdrich, for preemption of state laws under the Employee Retirement Income Security Act (ERISA). Though Pegram dealt with a fiduciary liability question, and not preemption specifically, the Court in arriving at its decision laid out a definition of the word "loan"--a word that is used in both the fiduciary liability section of ERISA and the preemption section. The Court's definition focuses upon the relationship between the managed care organization and the employer that hires it. The definition, however, excludes from the meaning of "plan" the relationship between the managed care organization and the health care providers it hires. Thus, this Note argues that according to Pegram, state laws that regulate the relationship between managed care organizations and health care providers, such as "any willing provider" laws, should not be preempted by ERISA. PMID- 11942338 TI - Fulfilling the bargain: how the science of ergonomics can inform the laws of workers' compensation. AB - In the last decade, cumulative trauma disorders have become a significant percentage of reported workplace injuries and litigated workers' compensation claims. Arising from the accumulated impact of daily work activities on the body, these injuries do not fall neatly within either the "accident" or "disease" categories which comprise workers' compensation laws. As a result, courts and legislatures have struggled to properly evaluate workers' compensation claims for these injuries. This Note looks at the legal treatment of cumulative trauma injuries in light of the "original bargain" of workers' compensation, where workers give up a tort remedy against their employers in exchange for guaranteed, but limited, compensation for work-related injuries. In doing so, this Note undertakes a comprehensive comparison of litigated cumulative trauma cases in the tort and workers' compensation systems. Ultimately, this Note argues that judges must use the original bargain as an interpretive less when deciding cumulative trauma cases, and points to ergonomics--the science of the workplace--as a significant new tool for determining whether such injuries are work-related. PMID- 11942339 TI - Resisting aspirin. PMID- 11942340 TI - Stents are the CADILLAC of care. Controlled Abciximab and Device Investigation to Lower Late Angioplasty Complications. PMID- 11942341 TI - Vitamins and donor hearts. PMID- 11942342 TI - Specific immune responses in changed gaseous environments. AB - The capacity of lymphoid cells to participate in immunity reactions was evaluated by blast transformation of lymphocytes under the influence of phytohemagglutinin. Blast transformation was measured by cytologic analysis and autoradiographic investigation of the rate of RNA synthesis in cells (tritiated uridin used as label). An analysis of the material taken from the three test subjects during the year-long experiment showed that various situations affected significantly the blast transformation level of lymphocytes. The reaction was substantially reduced 10 days after a simulated emergency situation which involved a change in the atmosphere, increase of physical load, etc. The level of blast transformation increased 1.5 to 2 months after the simulation, exceeding the average value, then to be normalized. Atmospheric variations appear to be one of the factors that may change the activity of lymphoid cells. A parallel experiment was performed in which three subjects lived 10 days in a hyperoxic enclosed environment (53% O2). They showed a considerable intensification of blast transformation (by 2.2-2.6 times) and pronounced activation of the RNA synthesis. Investigations give evidence that a long-term enclosure exerts an effect on the reactivity of the systems involved in the development of basic immune reactions. PMID- 11942343 TI - Changes in the microflora of man during long-term confinement. AB - Extended space missions may change the human normal microflora, including the intestinal flora. The bacterial composition of the intestinal microflora is an important factor in natural resistance to infection. During the year-long medico engineering experiment human intestinal microflora was thoroughly studied. It was necessary to collect more detailed information concerning the composition in healthy people. 83 microbiological tests of faeces of 27 healthy male subjects were carried out. It was found that spore-less anaerobic bacteria predominated in the fecal microflora, reaching 90% of total micro-organisms. Aerobic micro organisms were less than 6% of total micro-organisms. Long-term enclosure of men was shown to induce changes in the intestinal microflora which involved a sharp reduction of different microbial groups, and occasionally complete elimination of certain representatives of the intestinal flora. The composition tended to be simplified. A study of the isolated cultures of Cl. perfringens demonstrated an increased amount of strains with toxicogenic properties. Staphylococci were isolated from the test subjects during their long-term enclosure. The pharynx and nose of the three test subjects were examined and 1445 staphylococcal strains were isolated, of which 745 strains (51.5%) were coagulase producers. In 745 strains phage-type, coagulase, toxicity, hemolysins, lecitinase, fibrinolysin and DNA-ase were determined. Phage-typing of the isolated staphylococci revealed a microbial exchange between the men. Staphylococcal strains of the same phage-type (29/52) isolated during the experiment showed an increase of the toxin titre (from 1:40-1:80 to 1:640-1:1280) and development of additional pathogenic properties that were not found previously. An increase in beta-hemolysin and fibrinolysin production and an acceleration of DNA-ase activity. Experiments on mice demonstrated increased virulence of the staphylococcal strains isolated by the end of the experiment. The data are discussed from the point of view of the immunological responses of the human body. PMID- 11942344 TI - The ecology of micro-organisms in a closed environment. AB - Experimental data are presented which have a bearing on the susceptibility of astronauts to infectious diseases. These experiments include the observation of growth by two bacteria in Biosatellite 2, in which higher mean densities were attained than in earth-based controls. In addition, weightlessness combined with the special environment of a space vessel may affect the physiology of the astronauts. Earth-based studies in closed chambers were carried out under a variety of conditions with regard to pressure and oxygen content. One notable result was the transfer of micro-organisms from subject to subject. Comparative experiments were carried out in Antarctica and it was shown that the tests in closed chambers differed markedly from the Antarctica experience. The objectives and procedures of microbiological tests of the Gemini and Apollo programs are outlined. PMID- 11942345 TI - Life sciences and space research IX. Proceedings of the Open Meeting of Working Group 5 at the Thirteenth Plenary Meeting of COSPAR, Leningrad, USSR, 20-29 May, 1970. PMID- 11942346 TI - The importance of international activities to the work of the Food and Drug Administration's Center for Food Safety and Applied Nutrition. PMID- 11942347 TI - Reflections on delegation in the Chevron era. PMID- 11942348 TI - Protecting the balance of Hatch-Waxman: understanding the industry's new dynamics for the 21st century. PMID- 11942349 TI - Guidelines on gifts to physicians from industry: an update. PMID- 11942350 TI - WLF and the two-click rule: the First Amendment inequity of the Food and Drug Administration's regulation of off-label drug use information on the Internet. PMID- 11942352 TI - Regulatory and legal implications of reprocessing and reuse of single-use medical devices. PMID- 11942351 TI - Going 90 in a 55 M.P.H. speed zone: reprocessing of used single-use medical devices and the Food and Drug Administration's non-enforcement of the Food, Drug, and Cosmetic Act. PMID- 11942353 TI - An assessment of the regulations on medical devices in the European Union. PMID- 11942354 TI - Survival and mutability of Chlorella aboard the Zond vehicles. AB - Space effects on the viability and heredity of the unicellular green alga Chlorella were studied during the Soyuz 5 flight in earth orbit and in Zond 5, 6 and 7 on their earth-moon-earth flight. A culture of strain LARG-1 was exposed aboard the ship on mineral agar medium in the dark. The flight in earth orbit aboard the Soyuz-5 brought about a significant fall-off in cell survival and a gain in their mutability. There was a trend toward the percent growth of anomalies in autosporulation. The results obtained in experiments aboard the automatic stations of the Zond series were rather contradictory. The flight aboard Zond 6, like the earlier Zond 5, led to a statistically significant fall off in the cell survival (91.9 +/- 0.97% in the control; 80.2 +/- 1.78% in the test). The cell mutability remained unchanged; space had no effect on the cell viability in the experiment aboard Zond 7, but there was a trend toward antimutagenic action of the space factors (0.82 +/- 0.11% in the control; 0.57 +/ 0.06% in the test). Suggestions concerning the causes of these variances of result are reported. PMID- 11942355 TI - Cellular changes in wheat seedlings during orbital flight. AB - Wheat seedlings in weightlessness aboard NASA Biosatellite 2 differ from ground control seedlings in mitotic count, cell length and nuclear volume as well as in orientation, starch grain distribution, organ length and malformations previously reported to COSPAR. Dividing cells are fewer in roots of orbited seedlings than in erect or clinostat ground controls. The greatest difference is among cells in early prophase. Root cells, proximal to the zone of cell division are longer in flight seedlings than erect or clinostat ground controls. As the roots are the same length, greater elongation compensates for the reduced rate of cell division. Volume of interphase nuclei in all seedling organs is increased by orbital flight. In clinostat controls, only nuclei of primary roots increase in size. Between 58 and 65 hours of age, nuclear volume in erect 1 g coleoptiles decreases; in flight seedlings it increases. Simulated launch vibration, alone or followed by growth on the clinostat, increases nuclear volume in some roots, but coleoptile nuclei do not respond as in flight seedlings. Thus, at the cellular level, orbital flight cannot be exactly duplicated by the clinostat whether preceded by vibration or not. PMID- 11942356 TI - Post-flight histological analysis of turtles aboard Zond 7. AB - On the occasion of the flight of Zond 7 four turtles were included in the payload. A detailed histological analysis was carried out upon recovery of the subjects. No gross changes were observed, but the nuclei in the cells of many tissues decreased in volume by a statistically significant amount. This decrease is assumed to correspond to certain changes in function which adapted the animal to space flight conditions. No pathological changes were observed. PMID- 11942357 TI - Biosatellite 3: a physiological interpretation. AB - On 28 June 1969, a macaque monkey was boosted into orbit for a projected 30-day flight. It was aborted after 8 1/2 days and the monkey died 16 hours later. Data collected indicate: Weightlessness and the hypothermia acted to shift blood volume centrally; this provided a strong drive for the reduction of blood volume. Restraint, unusual vestibular sensations, and the continuing polydipsia all acted to disturb the central mechanisms affecting salt and water metabolism; it is probable that the function of the kidney was significantly affected and that an excessive amount of salt was lost. We can speculate that a serious electrolyte disturbance was superimposed on growing dehydration. Unpleasant vestibular sensations may have contributed to the high evaporative loss by autonomic disturbance, the whole problem being compounded and reinforced by the unnatural restraint to which the monkey was subjected. PMID- 11942358 TI - Experiments with micro-organisms and human cell cultures in the Zond 5 and Zond 7 flights. AB - Lysogenic strains of Escherichia coli were exposed to space conditions aboard the flight of Zond 5 and Zond 7. Space flight factors appeared to affect the state of episome systems of bacteria, as judged by data obtained with F-Lac+ donor cells which also carried genetic markers for threonine and leucine. Observations on phage induction are discussed and compared with results obtained aboard Biosatellite 2. A number of monolayer cultures of human cells (HeLa cells, fibroblasts, and A-1 cells) were repeatedly exposed to the space environment. In one instance, HeLa 19 cells increased in size after exposure to space conditions, a change which appeared to be genetically stable. HeLa 19 cells which were carried on six separate space flights showed a higher viability than corresponding cultures which were exposed only once aboard Zond 5. PMID- 11942359 TI - Temporal changes in reversible cerebral ischemia on perfusion- and diffusion weighted magnetic resonance imaging: the value of relative cerebral blood volume maps. AB - Using a transorbital approach we induced the temporal occlusion and reperfusion model in 18 cats. A vascular clamp was placed on the main trunk of the left middle cerebral artery (MCA) for 1 h. Diffusion- and perfusion-weighted MR images were obtained at 1, 3, 6 and 24 h after the clip was released. The cats were killed 24 h after reperfusion, and triphenyl tetrazolium chloride (TTC) staining was performed. After the relative cerebral blood volume (rCBV), time to peak enhancement (TTP) and apparent diffusion coefficient (ADC) maps had been acquired, ROIs were drawn on (1) the area of the infarct produced, (2) the area of high signal intensity on initial diffusion-weighted magnetic resonance imaging (DWI) but normal on TTC staining, e.g., salvaged parenchyma. The ratios of these areas to the normal contralateral cortex were calculated and compared with those of the areas of the final infarct and the salvaged parenchyma. Areas of final infarct showed a temporal increase of rCBV on 3 and 6-h imaging and a final depletion on 24-h imaging. A persistent decrease of ADC value and delayed TTP were observed. Salvaged parenchyma also showed increased rCBV after reperfusion until the last imaging comparing it to the final area of infarct (P < 0.05, 24-h rCBV). The initial decrease in the ADC and delayed TTP normalized on 24-h imaging. In conclusion, rCBV of 24-h imaging was the reliable parameter to predict final infarct. A combination of serial changes on DWI and perfusion weighted imaging (PWI) can predict ischemic penumbra and outcome. PMID- 11942360 TI - Diffusion-weighted MR imaging findings in carbon monoxide poisoning. AB - Diffusion-weighted MR imaging (DWI) of two patients with carbon monoxide (CO) poisoning demonstrated white matter and cortical hyperintensities. In one patient, the changes on the FLAIR sequence were more subtle than those on DWI. The DWI abnormality in this patient represented true restriction. In the second patient, repeated exposure to CO caused restricted diffusion. DWI may be helpful for earlier identification of the changes of acute CO poisoning. PMID- 11942361 TI - MRI demonstration of unsuspected intraventricular rupture of pyogenic cerebral abscesses in patients being treated for meningitis. AB - We report three patients with pyogenic meningitis who showed a partial response to medical treatment. MRI demonstrated an unsuspected brain abscess with intraventricular rupture, a possible cause of the partial response to treatment. PMID- 11942362 TI - MR and CT findings of craniopharyngioma during and after radiation therapy. AB - Long-term changes in craniopharyngioma treated with radiation therapy (RT) were investigated by computed tomography (CT) and/ or magnetic resonance (MR) imaging. Eight patients with craniopharyngioma were treated with incomplete resection or conservative surgical intervention followed by postoperative RT. The periods of tumor shrinkage were often long and varied (range: 6-68 months, mean: 29.1 months). Temporary enlargement of the solid component of a tumor usually occurs during RT and does not represent tumor progression. Cystic enlargement also occurs sometimes comparatively early after RT, and enlarged cysts often shrink with no treatment or with conservative treatment. These changes should be differentiated from tumor recurrence, with careful follow-up. After shrinkage, small solid or cystic nodules enhanced with contrast medium often remain. Long term follow-up is necessary to differentiate uncontrolled tumors from controlled tumors with imaging modalities. PMID- 11942363 TI - Astrocytoma of the pituitary gland (pituicytoma): case report. AB - A 34-year-old man presented with a 4-month history of visual obscuration. Magnetic resonance imaging showed a solid, discrete, contrast-enhancing pituitary mass with suprasellar extension. Surgery, which was performed via a transsphenoidal approach, disclosed the pituitary tumor to be a fibrillary astrocytoma (pituicytoma). This case report contains the clinical and neuroimaging features of this rare tumor of the neurohypophysis, which masqueraded as a pituitary adenoma. PMID- 11942365 TI - Transcortical mixed aphasia due to cerebral infarction in left inferior frontal lobe and temporo-parietal lobe. AB - We present a case of transcortical mixed aphasia caused by a cerebral embolism. A 77-year-old right-handed man was admitted to our hospital with speech disturbance and a right hemianopia. His spontaneous speech was remarkably reduced, and object naming, word fluency, comprehension, reading and writing were all severely disturbed. However, repetition of phonemes and sentences and reading aloud were fully preserved. Although magnetic resonance imaging (MRI) showed cerebral infarcts in the left frontal and parieto-occipital lobe which included the inferior frontal gyrus and angular gyrus, single photon emission CT revealed a wider area of low perfusion over the entire left hemisphere except for part of the left perisylvian language areas. The amytal (Wada) test, which was performed via the left internal carotid artery, revealed that the left hemisphere was dominant for language. Hence, it appears that transcortical mixed aphasia may be caused by the isolation of perisylvian speech areas, even if there is a lesion in the inferior frontal gyrus, due to disconnection from surrounding areas. PMID- 11942364 TI - Interobserver variability of magnetic resonance angiography in the diagnosis of carotid stenosis--effect of observer experience. AB - Magnetic resonance angiography (MRA) for determining the degree of carotid stenosis prior to carotid endarterectomy is attractive because it does not have the high morbidity associated with conventional intra-arterial angiography. We assessed the interobserver variability in the estimation of the degree of stenosis amongst observers of different experience. In a prospective study, consecutive patients with transient ischaemic attacks and symptomatic tight carotid stenosis shown by Doppler ultrasound underwent conventional intra arterial angiography and 2-D and 3-D time-of-flight MRA of the carotid bifurcations. The films of the processed MRA images were reviewed blind to other clinical and imaging data by eight observers of different levels of experience and coded for the presence and degree of stenosis. The stenosis on intra-arterial angiography was used as the reference standard. There was considerable variability between observers for estimation of the degree of stenosis on MRA. The observers' accuracy ranged from 41% (student) to 79% (experienced). From these estimations, excluding those of the student, it could be seen that up to 23% of patients who would have had an endarterectomy based on MRA should not have done so according to angiography, and up to 33% of patients who should have had an endarterectomy according to angiography would have inappropriately missed having an endarterectomy based on MRA results. Observer reliability of MRA processed images is not good, even in experienced hands. Using the source images and views of the circle of Willis might improve the accuracy, but a further study is required to assess this. PMID- 11942366 TI - Agenesis of internal carotid artery associated with congenital anterior hypopituitarism. AB - We report a rare case of unilateral agenesis of the internal carotid artery in association with congenital anterior hypopituitarism. The collateral circulation is supplied by a transsellar intercavernous anastomotic vessel connecting the internal carotid arteries. These abnormalities are well depicted on MRI and MRA. The agenesis of the internal carotid artery may explain the pathogenesis of some of congenital anterior hypopituitarism. PMID- 11942367 TI - Spinal radiological findings in nine patients with spontaneous intracranial hypotension. AB - Cranial magnetic resonance imaging (MRI) findings in spontaneous intracranial hypotension (SIH) are well known, while spinal studies have received less attention. Radiological spinal findings in nine patients with SIH are presented, looking for possible characteristic features. Five of the nine patients had histories of previous minor trauma, one of previous surgery; in three patients possible relevant preceding events were completely absent. All nine patients had cervical, seven thoracic, and four lumbar spine MRI studies; post-contrast studies were obtained in seven cases, MRI myelograms in five. Radioisotope myelocisternography was performed in four patients and myelo-CT in four. Epidural fluid collections were found in seven patients. In six cases the dural sac had collapsed, with a festooned appearance; intense epidural enhancement on post contrast studies demonstrated marked dilatation of the epidural venous plexus. In three cases an irregular root sleeve suggested a possible point of cerebrospinal fluid (CSF) leakage. Myelo-CT demonstrated the CSF fistula in two cases, radioisotope myelocisternography in three. The pattern of spinal abnormalities is different from that seen in cranial MRI for anatomical reasons: in the spinal canal the dura is not adherent to the bone; therefore, collapse of the dural sac and dilatation of epidural venous plexus occur, rather than subdural hematomas. In most cases the search for the dural tear is difficult. Radioisotope cisternography is probably the most sensitive examination for documenting the leakage of CSF out of the subarachnoid space; myelo-CT may precisely demonstrate the point of the CSF fistula, whereas MRI may only suggest it. PMID- 11942368 TI - Unusual origin of the artery of Adamkiewicz from the fourth lumbar artery. AB - The main arterial supply of the dorso-lumbar spinal cord is usually derived from a single anterior radiculo-medullary artery called the artery of Adamkiewicz and referred to as having a middle or low location. In some cases, the artery origin is higher, and a vessel which arises in the lower part of the region supplements the supply of the anterior spinal artery. In the literature, those arteries have been described as arising from L3 upwards, and have never been previously described angiographically, to our knowledge, below this level, although Suh and Alexander and Gililan have mentioned this eventuality. Of the 4,000 spinal cord angiographies performed in our institution, we report three cases in which the fourth lumbar artery flows into the anterior spinal artery of the conus medullaris. This anatomical variant may explain the sometimes devastating post operative neurological complications from a spinal cord infarction on surgery of the lumbar spine or the abdominal aorta below L3. PMID- 11942369 TI - Aberrant inferior petrosal sinus: unusual transvenous approach to the cavernous sinus. AB - Two patients with vascular lesions of the cavernous sinus (CS) undergoing endovascular management are reported. During transvenous embolization an unusually low termination of the inferior petrosal sinus (IPS) was observed. In both patients, we were able to catheterize the CS using this aberrant venous route. Knowledge of this variant can be crucial for a successful transvenous approach and treatment of vascular lesions involving the cavernous sinus. PMID- 11942371 TI - Intracranial lipoma with extracranial extension through foramen ovale in a patient with encephalocraniocutaneous lipomatosis syndrome. AB - We present CT and MR features of a large intracranial lipoma located in the left sphenocavernosal region in a patient with a history of spinal intradural lipoma excision. The lipomatous lesion surrounded the internal carotid artery and trigeminal nerve and extended extracranially into the infratemporal region through a smoothly dilated foramen ovale. PMID- 11942370 TI - Angioplasty and stenting of the cervical carotid bifurcation: report of a 4-year series. AB - Over a period of 46 months, 71 cervical carotid bifurcation stenotic lesions were treated by angioplasty and/or stenting. The stenosis was atherosclerotic in 61 cases, post-surgical or post-radiation in ten cases. The initial technical choice was that of primary stenting without cerebral protection; this attitude has been modified in recent cases by use of a protection balloon or filter. A technical success rate of 98% was achieved (71 cases treated out of 73 attempted). From the angiographic point of view, in all cases good restoration of the luminal diameter was obtained. There were three completely regressive minor strokes (4.2%), and one major stroke (1.4%). A follow-up of at least 1 year is available for 50 patients; a restenosis of less than 50% occurred in four (8%). In conclusion, angioplasty and stenting of the cervical carotid bifurcation is feasible in relatively safe conditions. However, analysis of the case complicated by a major stroke and the evolution of protection devices in recent years suggest the use of such devices for brain protection. The most suitable condition for endovascular treatment is considered to be an intermediate stenosis, i.e. with a residual lumen for safe passage of the devices used, without need for dangerous friction on plaque surfaces. The most dangerous phase of the treatment is dilation of the plaque by the balloon; deployment of an oversized stent without dilation is desirable whenever possible. PMID- 11942372 TI - Magnetic resonance imaging of metastatic disease to the brain with gadobenate dimeglumine. AB - Seventy-four patients with one to eight proven intraaxial brain metastases received a total cumulative dose of 0.2 mmol/kg bodyweight gadobenate dimeglumine, administered as sequential injections of 0.05, 0.05 and 0.1 m mol/kg over a 20-min period. MR imaging was performed before the first administration (T2- and T1-weighted sequences) and after each injection of contrast agent (T1 weighted sequences only). Quantitative assessment of images revealed significant (P <0.01) dose-related increases in lesion-to-brain (L/B) ratio and percent enhancement of lesion signal intensity. Qualitative assessment by two independent, blinded assessors revealed additional lesions in 22%, 25% and 38% (assessor 1) and 29%, 32% and 34% (assessor 2) of patients after each cumulative dose when compared with combined T1- and T2-weighted pre-contrast images. Significantly more lesions (P < 0.01) were noted by both assessors after the first injection and by one assessor after each subsequent injection. For patients with just one lesion observed on unenhanced T1- and T2-weighted images, additional lesions were noted in 12%, 16% and 28% of patients by assessor 1 following each dose and in 24%, 27% and 30% of patients by assessor 2. Contemporaneously, diagnostic confidence was increased and lesion conspicuity improved over unenhanced MRI. For patients with one lesion observed after an initial dose of 0.05 mmol/kg, additional lesions were noted by assessors 1 and 2 in 9.1% and 11.8% of patients, respectively, after a cumulative dose of 0.1 mmol/kg and in a further 9.1% and 5.9% of patients, respectively, after a cumulative dose of 0.2 mmol/kg. No safety concerns were apparent. PMID- 11942373 TI - Characteristic MRI findings in multiple system atrophy: comparison of the three subtypes. AB - We reviewed MRI findings in 29 patients with probable multiple system atrophy (MSA) to see whether there were common and or less common neuroradiological findings in the various clinical subtypes. We divided the patients into three clinical subtypes according to initial and predominant symptoms: 14 with olivopontocerebellar atrophy (OPCA), eight with the Shy-Drager syndrome (SDS) and seven with striatonigral degeneration (SND). The patients showed atrophy of the brain stem and cerebellum, high signal on T2-weighted images of the base of the pons and middle cerebellar peduncles, high and low signal on T2-weighted images of the putamen and atrophy of frontal and parietal lobes. The degree of atrophy of the middle cerebellar peduncle and cerebellum was greater in OPCA patients and a high-signal lateral rim to the putamen more frequent in SND. However, all findings were observed in all subtypes, and the degrees of atrophy of the putamen and pons and the frequency of high signal in the base of the pons were similar in the subtypes. We also found atrophy of the cerebral hemispheres, especially the frontal and parietal lobes, but its degree was not significantly different in the various subtypes. Our findings suggest that, although MSA can be divided clinically into three subtypes, most of the features on MRI are common and overlap in the subtypes, independently of the clinical presentation. PMID- 11942374 TI - Magnetization transfer imaging diagnosis of intracranial tuberculomas. AB - Tuberculomas (TB) can mimic brain tumors and abscesses. We performed prospective magnetization transfer ratio (MTR) analysis on 60 patients with 52 intracranial TB, 13 pyogenic abscesses and 65 tumors to determine the efficacy of magnetization transfer (MT) imaging in the differential diagnosis of brain tumors and infection. The nonenhancing cores of TB had significantly higher (P=.026) MTR (0.14+/-0.29) than necrotic components of high-grade gliomas (-0.19+/-0.22). The MTR of cores of TB were also higher than those of the cystic areas of low-grade gliomas (-0.53+/-0.32), benign (-0.09+/-0.21) and malignant (-0.07+/-0.25) tumors, and abscesses (-0.03+/-0.13), but the differences were not significant because of the small number of tumors and abscesses. There was also no significant difference between the MTR of abscesses, malignant and benign tumors. Using the criteria of MTR of necrotic center > 0.14 (mean MTR of TB) for diagnosing TB, MTR <-0.03 (mean MTR of abscesses) for diagnosing tumors and MTR between these values for diagnosing abscesses, MTI had diagnostic sensitivity of 68.42%, specificity of 80.49%, and accuracy of 76.67%. The improved diagnostic accuracy of MRI with the addition of MTR analysis from 86.67% to 91.67% and from 85.71% to 87.50% for both radiologists respectively was not significant. MTR analysis helped us to differentiate solitary TB or abscess from low-grade glioma in five patients and to diagnose multiple TB, abscesses, and metastases in four. PMID- 11942375 TI - Differentiation of radiation necrosis from tumor progression using proton magnetic resonance spectroscopy. AB - We report on a young woman who was treated by stereotactic radiotherapy for recurrence of an initially resected low-grade astrocytoma. MRI follow-up examination 7 months after radiotherapy showed a gadolinium-DTPA-enhancing mass lesion indicative of high-grade tumor progression. This assumption was also supported by positron emission tomography with [2-18F]fluoro-2-deoxy-D-glucose (FDG-PET). In contrast, proton MR spectroscopy (1H-MRS) indicated radiation necrosis, which was confirmed histopathologically in surgical specimens. Subsequent follow-up examinations up to 19 months after surgery showed no evidence of tumor recurrence. PMID- 11942376 TI - Sequential MRI in a case of Creutzfeldt-Jakob disease. AB - A 48-year-old man suddenly developed clinically and electroencephalographically nonspecific dementia. On MRI sequences, only diffusion-weighted images (DWI) of the cortex were unequivocally pathological. Obvious atrophy and basal ganglia signal changes appeared only 9 months after the onset. Brain biopsy confirmed Creutzfeldt-Jakob disease (CJD). In rapidly progressive dementia, we recommend DWI for early diagnosis of CJD. PMID- 11942377 TI - Progressive multifocal leukoencephalopathy limited to the brain stem. AB - Progressive multifocal leukoencephalopathy (PML) is a subacute demyelinating slow virus encephalitis caused by the JC polyomavirus in 2-5% of patients with AIDS. MRI typically shows multiple lesions in the cerebral hemispheres. We present a rare case of rapidly evolving and lethal PML with a severe bulbar syndrome and spastic tetraparesis in a patient with AIDS. MRI showed high-signal lesions on T2 weighted images confined to the brain stem, extending from the medulla oblongata to the midbrain. JC virus polymerase chain reaction in cerebrospinal fluid was positive, and neuropathology showed the findings of PML. This case was also notable because of the rapid progression despite improved immune status with antiretroviral therapy. PMID- 11942378 TI - Proton magnetic resonance spectroscopy of neurocytoma outside the ventricular region--case report and review of the literature. AB - Central neurocytoma is classically considered as an intraventricular benign tumour, largely based on data from small retrospective series and single case reports. We report on a 16-year-old girl who suffered from a large parietooccipital tumour that was diagnosed histologically as central neurocytoma. The features of CT, MRI and proton MR spectroscopy studies are discussed. This is the first report on spectroscopic findings in a case of extraventricular neurocytoma. As well as elevated choline (Cho), the tumour spectrum showed strongly decreased N-acetylaspartate (NAA). NAA is assumed to be produced in mature neurons, and we therefore expected to find high amounts of NAA in this well-differentiated tumour, which was histologically composed of mature neuronal tissue. This observation leads to the conclusion that even the highly differentiated cells of neurocytomas are too immature to produce NAA. PMID- 11942379 TI - Transient global amnesia: increased signal intensity in the right hippocampus on diffusion-weighted magnetic resonance imaging. AB - We report on a patient with pure transient global amnesia (TGA) whose magnetic resonance imaging (MRI) demonstrated a small region of increased signal intensity in the right hippocampus on diffusion-weighted imaging (DWI). DWI was sensitive and useful for evaluating the early stage of TGA and might help to explain the pathophysiology of TGA. PMID- 11942380 TI - Primary CNS lymphoma in the spinal cord: clinical manifestations may precede MRI detectability. AB - We report a 36-year-old woman whose first MRI after developing symptoms of cervical nerve root irritation was normal. Eight months later, after progression to tetraparesis and demonstration of an extensive contrast-enhancing intramedullary lesion giving high signal on T2-weighted images, the diagnosis of lymphoblastic non-Hodgkin's lymphoma was made by biopsy. The lesion responded for a short time to corticosteroid medication, but later rapidly extended to the epidural soft tissues, probably due to a cerebrospinal fluid fistula. Despite radio-and chemotherapy, the patient survived only 7 months from histological diagnosis and 15 months from the onset of symptoms. PMID- 11942381 TI - Concurrent spinal cord and vertebral bone marrow radionecrosis 8 years after therapeutic irradiation. AB - Concurrent radionecrosis within the spinal cord and the bone marrow at the same thoracic level was observed 8 years after localized therapeutic irradiation in a patient who had undergone repeated cycles of radiotherapy, glucocorticoid treatment, and chemotherapy for a non-Hodgkin's lymphoma. Mechanisms combining radiotoxic potentialization by glucocorticoids/alkylating agents and delayed radiation-induced vasculitis involving the common arterial pathways to the spinal cord and to the vertebrae were speculated to have acted in a synergistic way. PMID- 11942382 TI - A case of aseptic vertebral necrosis in the context of metastatic lumbar disease. AB - Aseptic bone necrosis has many well recognized etiologies: caisson disease, corticosteroids, alcohol, Gaucher's disease and sickle cell anemia being a few of a long list. Little attention has been paid to metastatic disease as a possible cause and we were unable to find reference in the literature to this association. We present imaging features of metastatic Ewing's sarcoma involving the vertebral spine, which, in certain aspects, mimics other well-known entities of this region, such as Kummell's and Scheuermann's diseases. PMID- 11942383 TI - A rare cause of cervical spinal stenosis: posterior arch hypoplasia in a bipartite atlas. AB - We describe CT and MRI of a previously unreported combination of atlantoaxial anomalies consisting of posterior arch hypoplasia in a bipartite atlas with an os odontoideum, in a 30-year-old woman presenting with neck and left arm pain. MRI showed the os odontoideum, marked stenosis of the spinal canal at the level of the atlas, with cord compression and evidence of myelopathy. CT revealed a bipartite atlas with midline clefts in anterior and posterior arches, thickening in the anterior arch and hypoplasia of the posterior arch with incurving of both hemiarches. Flexion and extension radiographs demonstrated atlantoaxial instability. PMID- 11942384 TI - Spinal epidural en-plaque meningioma with an unusual pattern of calcification in a 14-year-old girl: case report and review of the literature. AB - A 14-year-old girl who presented with a severe sensory-motor-sphincter syndrome was found to be harboring an epidural tumor situated posteriorly in the spinal canal from C5 through C7 levels. The mass had computerized tomography and magnetic resonance imaging features suggesting an unusual stratified architecture, with a conspicuous highly calcific component firmly adherent to the dura and a non-calcific mass surrounding it posteriorly and laterally. Although meningiomas have a low incidence in the first two decades of life, and in the spine they rarely have entirely extradural location at any age, a meningioma was suspected. Intraoperative biopsy confirmed the tumor to be benign, and careful total resection including the whole large dural implant was carried out; the wide dural defect was grafted with fascia lata. A meningothelial meningioma with a largely calcified psammomatous component was diagnosed. The girl made a complete recovery, and is tumor-free 9 years postoperatively. Outcome from surgery for spinal meningiomas can be good, despite the severity of the preoperative condition; however, enplaque and highly calcific tumors still bear a poorer prognosis. Complete resection is mandatory in children, in whom they are extremely rare. Modern imaging techniques help in making a correct initial diagnosis and optimizing surgery in order to provide good results, even in more challenging cases. PMID- 11942385 TI - Performance of long J-shaped coils in large and giant intracranial aneurysms: an in vitro study. AB - We evaluated the performance of long straight coils (Detach-18 J-shaped coils) in large and giant in-vitro aneurysms. The coils consisted of a distal semicircular part 7 or 15 mm in diameter and a proximal 70 cm straight part having three types of stiffness: soft, standard, and stiff. We first investigated the ease of passage through a microcatheter in a realistic cerebral vessel model. Second, we made silicone models simulating lateral-type aneurysms of various spherical dome diameters (12, 15, 20 and 30 mm) and neck sizes (3-10 mm; dome-to-neck ratio approximately 3:1) and connected them to a pulsating circulatory pump. We evaluated the anchoring and folding patterns of the coils, stability of the microcatheter and coils in the aneurysm, and smoothness of delivery and retrieval of coils. Third, we compared the conformability of a coil in a large, irregular aneurysm with that of a spiral coil. The long J-shaped coils were easily advanced and retrieved through a microcatheter in a tortuous vessel model. In 12 mm spherical aneurysms, each coil made a complex framework, and knot formation or damage to the coil during withdrawal was often observed. In 15 mm aneurysms, the coils were all easily delivered and retrieved; standard-stiffness coils adapted best to this size. In 20 mm aneurysms, frameworks were less complex but still good with a standard or stiff coil, but those with soft coil were unstable and changed significantly within 3 min of detachment due to gravity and pulsatile flow. In 30 mm aneurysms, soft coils filled only the lower part during introduction, whereas a stiff coil still made a favourable framework. Conformability of a long J-shaped coil was superior to that of a spiral coil in a large, irregular aneurysm. Long J-shaped coils conform well to various configurations of large and giant aneurysms and can shorten procedures since a larger implant volume can be delivered with a single coil. Their principle of action may promote more favourable occlusion of irregular and large aneurysms by improved conformability and stability. PMID- 11942386 TI - Natural history of chondroid skull base lesions--case report and review. AB - Long-term follow-up reports on chondroid lesions of the skull base are rarely presented in the literature. There are virtually no data on natural growth rates of these tumors based on MRI obtained over a period of 10 years or longer. We followed a patient who has had such a lesion for more than 12 years. A non progressive, slight abducens palsy has been the only associated symptom so far. Even though the patient was operated on for an additional intracranial arterio venous malformation, clinical features and chromosomal testing excluded Maffucci's syndrome. The MRI follow-up in this case provides an extraordinary perspective on the natural history of chondroid skull base tumors. PMID- 11942387 TI - Bilateral orbital pseudotumour due to Ormond's disease: MR imaging and CT findings. AB - We present the MRI and CT findings in a 43-year old patient with bilateral orbital pseudotumour due to Ormond's disease. PMID- 11942388 TI - Diagnosing dural metastases. PMID- 11942389 TI - Vibrational spectra of CCl4: isotopic components and hot bands. Part I. AB - Room temperature (295 degrees K) infrared spectra of CCl4 have been measured covering the 4000-200 cm(-1) region. Bands ascribable to the isotopic components in the first overtone of the asymmetric deformation (2v4) mode and the combination of the symmetric and the asymmetric deformation (v2 + v4) mode regions are observed in agreement with the percentage distribution of various isotopes. From these observations, the first ever estimate of the isotopic band positions in the regions of the v4 and v2 modes at room temperature are obtained. Successful assignment of the isotopic bands in the asymmetric stretching (v3) and combination of the symmetric stretching and asymmetric deformation (v1 + v4) mode regions has been achieved and realistic values of all the v3/(v1 + v4) mode Fermi resonance parameters are obtained. The experimental results suggest that the transitions, where the initial levels are not the ground state, contribute significantly and give rise to a very complicated vibrational spectrum for CCl4 vapour. PMID- 11942390 TI - Characterization of crude oils using fluorescence lifetime data. AB - The average fluorescence lifetimes of nine North Sea crude oils with API gravities of between 20 and 51 were measured using a modular, filter based, instrument developed in-house. Two pulsed light emitting diode (LED) excitation sources (460 and 510 nm) were used to excite fluorescence, the lifetime of which was measured at a range of emission wavelengths. Fluorescence lifetimes were found to vary from 1.8 to 8.2 ns with confidence intervals of +/- 0.11 ns. The average lifetimes at all emission wavelengths were linearly correlated with API gravity and with aromatic concentration with the best results being obtained with the 460 nm excitation source. Predictive models with an accuracy of +/- 7.6 API degrees were generated using partial least-squares methods from average fluorescence lifetimes measured at an emission wavelength of 500 nm using 460 nm excitation. A better correlation was found between the aromatic concentration of the oils and the ratio of the average fluorescence lifetimes at measured at 550 and 650 nm using 460 nm excitation. This led to a quantitative model with an accuracy of +/- 5.4% for aromatic concentration. PMID- 11942391 TI - Physical properties and spectra of IO, IO- and HOI studied by ab initio methods. AB - Structure and properties of the IO, IO- and HOI species, which are of potential importance for the ozone destruction catalytic cycle in the troposphere, have been calculated together with the EPR, NMR and UV-visible spectra by ab initio methodology with account of spin-orbit coupling (SOC) effects. Multi configuration self-consistent field calculations with linear and quadratic response techniques and the multi-reference configuration interaction method have been employed. Photodissociation of these species, crucial for the catalytic ozone-destruction cycle, is critically reviewed and analyzed. Calculations predict that the singlet-triplet (S-T) transition to the lowest triplet state (X1 A' --> 3A'') should be responsible for the weak long-wavelength tail absorption (approximately 450-560 nm) and photodissociation of the HOI molecule. The second, more intense, band around 400 nm is produced by two overlapping S-S and S-T transitions. In order to check this assignment of the HOI photodissociation the isoelectronic IO- anion and IO radical have been studied by the same methods. Comparison with the EPR spectrum of the IO radical indicates that the methods are reliable which gives credit to the accuracy of the HOI spectral interpretation. NMR spectra of HOI and IO- molecules and some other properties are calculated for the first time. PMID- 11942392 TI - Anharmonic vibrational spectra of hydroxylamine and its 15N, 18O, and deuterium substituted analogs. AB - Vibrational self-consistent field (VSCF) and correlation-corrected vibrational self-consistent field (CC-VSCF) methods were used to compute the anharmonic frequencies of fundamentals, overtones, and combination transitions of natural abundance hydroxylamine, 15NH2OH, NH2(18)OH, ND2OD, ND2OH, and NH2OD isotopomers at second order Moller-Plesset perturbation theory (MP2) in basis sets of triple zeta quality. Frequencies of the fundamental transitions observed in the gas phase spectrum were reproduced by CC-VSCF treatment within 20 cm(-1) in TZV(d,p) and TZV(2d,2p) basis sets, and the change of basis set composition had only minor effect on the frequencies of the computed fundamentals. CC-VSCF computed wave numbers of overtone and combination transitions were typically within 1-40 cm(-1) of the gas phase band positions, except for those resulting from multiple excitations of v2, v3, and v7 fundamentals, because of a strong mutual coupling between these modes. Integral transition intensities calculated at MP2 level closely followed those of experimental spectrum, including intensity decrease in v1, 2v1, 3v1 progression, and 30-fold intensity increase of 2v8 in respect to that of v8 fundamental. The frequency of the OH torsional fundamental was found to be strongly dependent on the mode-mode interaction potential among v9 and v1, v7, v2, v4, v5 modes. Band shifts resulting from 15N, 18O and complete 2H substitutions were reproduced almost quantitatively by CC-VSCF computation in TZV(d,p) basis. Computed anharmonic isotope frequency shifts were different from those obtained in the harmonic approximation and no scaling procedure seemed capable of performing their interchange. PMID- 11942393 TI - A reinvestigation of the molecular structures, vibrations and rotation of methyl group in o-methylaniline in S0 and S1 states studied by laser induced fluorescence spectroscopy and ab initio calculations. AB - The UV fluorescence excitation and dispersed fluorescence spectra of a jet-cooled o-methylaniline have been obtained for the S1 <-- S0 transition, in which some of the bands have been observed and assigned for the first time. The origin of the electronic transition appears at 34,328.4 cm(-1). It was found that the spectra exhibit an important feature corresponding to the internal rotation of the methyl group in the electronic ground and excited states. Ab initio calculations at MP2/6-31 + G* and CIS/6-31 + G* show that the optimised structure of o methylaniline in the ground state is not planar with the amino group having sp3 hybridation-like character due to the existence of lone paired electrons on the N atom. Upon electronic excitation, the C-N bond exhibits a partial double character, as in the case of other aniline derivatives. PMID- 11942394 TI - Gas-phase generation and infrared spectroscopy of metastable OP-SCN molecule. AB - An attempt has been made to generate the metastable phosphorus (III) oxythiocyanide molecule, OP-SCN for the first time by an on-line process using the vapor flow of phosphorus (III) oxychloride, OPCl passed over heated AgSCN at 520 degrees C. The products have been characterized by their IR spectra; values for v1 of 2055 cm(-1) (C[triple bond]N stretch), v2 of 1365 cm(-1) (O=P stretch), v3 of 720 cm(-1) (C-S stretch) and v7 of 680 cm(-1) (P-S stretch) have been obtained. PMID- 11942395 TI - Partial donor-donor energy migration (PDDEM) as a fluorescence spectroscopic tool for measuring distances in biomacromolecules. AB - A theoretical model is presented, tested and applied for determining the rates of energy migration and distances within pairs of chemically identical fluorophores, so-called donors (D), which are exposed to different physical properties. The model is a general extension of the recently developed donor-donor energy migration (DDEM) model [J. Chem. Soc., Faraday Trans. 92 (1996)1563; J. Chem. Phys. 105 (1996) 10896] that applies to examining structure-function of biomacromolecules, such as proteins. Most fluorescent groups of the same kind incorporated at different positions (alpha and beta) in a macromolecule exhibit shifts of the absorption and/or emission spectra, as well as different relaxation rates of the photophysics. As a consequence, the energy migration between the D(alpha) and D(beta) groups will be partially reversible. We refer to this case, as the partial donor-donor energy migration (PDDEM). The models of PPDEM presented can be used for analysing time-resolved fluorescence relaxation, as well as fluorescence depolarisation experiments. To explore the limitations of the PDDEM model, we have generated and re-analysed synthetic data that mimic time correlated single photon counting (TCSPC) experiments. It was found that slow and fast rates of energy migration are most accurately recovered from the fluorescence relaxation and the depolarisation experiments, respectively. At comparable transfer and fluorescence rates, both kinds of experiments are equally useful. Real experiments on PDDEM were performed on an asymmetrically quenched bichromophoric molecule (1,32-dihydroxy-dotriacontane-bis-(Rhodamine 101) ester), that spans across the lipid bilayer of a vesicle. The depolarisation data were analysed by the PDDEM model and provide a distance between Rhodamine 101 groups, which agrees with independent studies. PMID- 11942396 TI - Raman and FTIR spectra of [Cu(H2O)6](BrO3)2 and [Al(H2O)6](BrO3)3 x 3H2O. AB - Raman and FTIR spectra of [Cu(H2O)6](BrO3)2 and [Al(H2O)6](BrO3)3 x 3H2O are recorded and analyzed. The observed bands are assigned on the basis of BrO3- and H2O vibrations. Additional bands obtained in the region of v3 and v1 modes in [Cu(H2O)6](BrO3)2 are due to the lifting of degeneracy of v3 modes, since the BrO3- ion occupies a site of lower symmetry. The appearance v1 mode of BrO3- anion at a lower wavenumber (771 cm(-1)) is attributed to the attachment of hydrogen to the BrO3- anion. The presence of three inequivalent bromate groups in the [Al(H2O)6](BrO3)3 x 3H2O structure is confirmed. The lifting of degeneracy of v4 mode indicates that the symmetry of BrO3- anion is lowered in the above crystal from C3v to C1. The appearance of additional bands in the stretching and bonding mode regions of water indicates the presence of hydrogen bonds of different strengths in both the crystals. Temperature dependent Raman spectra of single crystal [Cu(H2O)6](BrO3)2 are recorded in the range 77-523 K for various temperatures. A small structural rearrangement takes place in BrO3- ion in the crystal at 391 K. Hydrogen bounds in the crystal are rearranging themselves leading to the loss of one water molecule at 485 K. This is preceded by the reorientation of BrO3- ions causing a phase transition at 447 K. Changes in intensities and wavenumbers of the bands and the narrowing down of the bands at 77 K are attributed to the settling down of protons into ordered positions in the crystal. PMID- 11942397 TI - A quantum-chemical study of the structure, vibrations and SiH bond properties of disilylamine, NH(SiH3)2. AB - Quantum-chemical calculations at HF, MP2 and B3LYP levels with 6-31G* and 6 311G** basis sets are reported for disilylamine, NH(SiH3)2. The equilibrium structure is found to vary with both level and basis set, all but one of the structures exhibiting a small lack of planarity of the HNSi2 system. The barrier to inversion, however, is found to be very low, at most 38 cm(-1). Vibration frequencies and intensities are calculated. The frequencies are scaled, where possible, either using updated infrared data or with the aid of factors transferred from N(CH3)(SiH3)2. Unobserved frequencies due to the v(s)NSi2, deltaNSi2 and delta(perpendicular)NH modes are predicted near 610, 210 and 360 cm(-1), respectively. The lower silyl torsion lies below 40 cm(-1). The appearance of a single broad vSiH band in gas-phase samples of both NH(SiH3)2 and NH(SiH3)(SiD3) is suggestive of signal averaging due to internal rotation. The frequencies v(is)SiH, infrared intensities and Raman scattering activities of the bands due to an isolated SiH bond in an otherwise deuterated species are calculated and correlated with the torsional angle of this bond and with the Mulliken charge on the hydrogen atom. The strength of the bond is a minimum, and the infrared intensity and Raman scattering activity are maxima, when the bond direction is roughly orthogonal to the skeletal plane. A major part of the frequency and intensity variations is attributed to n(p)(N)-sigma*(Si-H)) hyperconjugation which, NBO calculations show, reaches a maximum for this conformation. However, systematic smaller variations are found for SiH bonds lying in the skeletal plane, which reflect the proximity of the other silyl group and only partly correlate with Mulliken charge. vSiH-vSiH interaction force constants, f', are calculated for pairs of SiH bonds in different silyl groups and compared with the corresponding dipole-dipole potential energy, the latter calculated using a classical treatment of the interaction between point dipoles arising from delta mu/delta r for the SiH bonds involved. The gradient of the correlation is very close to that expected from the theory, but a negative intercept indicates the presence of additional factors. PMID- 11942398 TI - Laser excitation spectrum and spectroscopic potential parameters of Cd2 molecule in the 1u(5(3)P2) energy state. AB - Excitation spectrum of Cd2 van der Waals complex was observed in a supersonic free-jet expansion beam. Cadmium dimers were seeded in argon environment (a carrier gas) while a pulsed dye laser served as an excitation light source. Well resolved structures of vibrational bands arising from transition between 1u(3pi(u)) and X0g+ (1sigmag+) states were observed. Using a Birge-Sponer method for analyzing the vibrational excitation spectrum, the first-time determination of the spectroscopical potential parameters of the 1u molecular state was performed. We obtained D'e = 723 +/- 10 cm(-1), omega'e = 28.9 +/- 1.0 cm(-1), omega'e x'e = 0.260 +/- 0.002 cm(-1) and R'e = 3.93 +/- 0.05 A. The latter value was estimated with the help of the computer-simulation of the spectrum. The results are compared with recent results of ab initio calculation of Czuchaj et al. (Chem. Phys. Lett. 225 (1994) 233) and demonstrate a reasonable agreement. PMID- 11942399 TI - Vibrational spectroscopic study on pyrimidine metal(II) tetracyanometalate complexes. AB - The results of an infrared and Raman spectroscopic study are reported for seven new metal(II) pyrimidine tetracyanonickelate complexes, M(pyr)2Ni(CN)4 [where (pyr) = pyrimidine; M = Mn, Fe, Co, Zn, Ni, Cu or Cd] and an IR spectroscopic study is presented for new cadmium pyrimidine tetracyanometalate complex, Cd(pyr)2Cd(CN)4. The spectral data suggest that the first seven compounds belong to the Hofmann-type and the last compound belongs to the Hofmann-Td-type of complexes. PMID- 11942400 TI - FT-IR observation of covalent labelling of lysozyme crystals by organometallic complexes of transition metals. AB - Electrophilic complexes of iron and chromium which have been reported to react with proteins in solution have been reacted with hen-egg white lysozyme (HEWL) in both the solution and crystal phases under similar pH and buffer conditions. This work was carried out with a view to developing novel side-chain selective heavy metal derivatives for protein X-ray crystallographic studies. Reaction of HEWL with a tricarbonyldienyliron cation (1) in aqueous solution led to modification of the sole histidine residue with concurrent reversible modification of other protein residues. Reaction of (1) with crystalline HEWL showed no covalent binding and only a build up of a hydrolysis product in the water channels of the crystal was observed. Reactions with a series of tricarbonylarylchromium pyrylium salts (2) led to the formation of stable covalent HEWL derivatives in solution. Chromatographic and IR spectroscopic studies showed that binding took place specifically at the epsilon-amino group of lysine residues to give a series of mono- and di-substituted products. When crystals of HEWL were soaked with the chromium reagents covalent binding to some of the lysine residues was also observed. In contrast, HEWL crystals which had their lysine side chains disabled did not bind any of the chromium reagents. PMID- 11942401 TI - Spectroscopic characterisation of indium(III) chloride and mixed ligand complexes. AB - Acidic solutions of InCl3 plus InBr3 contain mixed halide complexes and solvent extraction yields the four-coordinate anions InCl(n)Br(4 - n)-, whose v(In-Cl) and v(In-Br) modes are reported. Two solid products of InCl3 with 15-crown-5 have been identified; crystallisation from aqueous HCI gave a molecular adduct containing InCl3(H2O)2, whereas the ionic complex [InCl2(15-crown-5)][InCl4] was produced under anhydrous conditions. The vibrational spectra are assigned. PMID- 11942402 TI - Infrared spectroscopy of the mineralogy of coprolites from Brean Down: evidence of past human activities and animal husbandry. AB - The mineralogy of 11 concretions from the Bronze Age settlement horizons at Brean Down near Weston-super-Mare, Somerset, UK, has been examined by infrared spectroscopy. The concretions are found to contain calcite and apatite and, in some cases, quartz. Four further concretions from the later Iron Age Meare Village, soil samples from Brean Down and mineralised samples of known faecal origin from a cesspit within the Tudor Merchant's house in Tenby have been similarly examined. It is found that all samples contain calcite, but only the concretions and the Tenby cesspit samples contain apatite. None of the soil samples contain apatite, although these are relatively high in quartz. This suggests that the concretions are coprolites and that the apatite has a biological origin in small bone fragments. The infrared study is backed up by scanning electron microscopy which confirms the presence of phosphorus in the coprolite samples and shows a morphology suggestive of the presence of bone fragments; it is likely, therefore, that the coprolites result from a carnivore- most probably from dogs. The findings show the usefulness of infrared spectroscopy for the rapid identification of mineralised coprolitic material from archaeological sites. PMID- 11942403 TI - Infrared spectroscopy of goethite dehydroxylation: III. FT-IR microscopy of in situ study of the thermal transformation of goethite to hematite. AB - Fourier transform infrared microscopy has been used to investigate in situ dehydroxylation of goethite to form hematite. The characterisation was based on the behaviour of hydroxyl units, which were observed in the hydroxyl stretching and hydroxyl deformation and water bending regions, and the Fe-O vibrations of the newly formed hematite during the thermal dehydroxylation process. Two hydroxyl stretching modes (v1 and v2), and three bending (V(bending-1, 2, 3)) and two deformation (V(deformation-1, 2)) modes were observed for goethite. The characteristic vibration at 916 cm(-1) was observed together with the residuals of the v1 and v2 bands in hematite spectrum. The structural transformation between goethite and hematite through thermal dehydroxylation was interpreted in order to provide criteria that can be used for the characterisation of thermally activated bauxite and their conversion to activated alumina phases. PMID- 11942404 TI - Two rotors potential scans and vibrational assignments for dihalomethylsulfonyl isocyanates. AB - The conformational behavior and structural stability of dichloro and difluoromethyl-sulfonyl isocyanates were investigated by quantum mechanical DFT and ab initio calculations. The 6-311 + + G** basis set was employed to include polarization and diffuse functions in the calculation at B3LYP and MP2 levels. The molecules were found to exist in a mixture of trans-gauche and gauche-gauche conformations at ambient temperatures. From the calculations the isocyanate NCO moiety was predicted to nearly eclipse one of the sulfony S=O bonds in the two stable conformers of both molecules. The potential scans for the rotations of the two NCO and CX2H rotors were calculated from which the rotational barriers could be estimated. The vibrational frequencies, potential energy distributions, IR intensities as well as depolarization ratios were calculated. PMID- 11942405 TI - Extension of the density functional derived scaled quantum mechanical force field procedure. AB - The density functional derived scaled quantum mechanical (SQM) force field method of Rauhut and Pulay [J. Phys. Chem. 99 (1995) 3093] has been extended. The original procedure (employing B3-LYP/6-31G* computations and 11 transferable scale factors for the different kinds of internal coordinates) was capable to reproduce the vibrational fundamentals of 31 simple organic (H, C, N, O) molecules with a total mean deviation of about 13 cm(-1). The present Density Functional Theory based SQM force field method is an extension of the original one: with the help of 20 transferable scale factors can reproduce the fundamentals of 20 inorganic, organic and organosilicon molecules containing nonmetallic first and second-row atoms with a total mean deviation of 10.8 cm( 1). The transferability and reliability of the new set of scale factors are demonstrated on the examples of the a priori SQM vibrational spectra of cis and gauche-cyclopropylchlorosilane. PMID- 11942406 TI - Telomere and telomerase in oncology. AB - Shortening of the telomeric DNA at the chromosome ends is presumed to limit the lifespan of human cells and elicit a signal for the onset of cellular senescence. To continually proliferate across the senescent checkpoint, cells must restore and preserve telomere length. This can be achieved by telomerase, which has the reverse transcriptase activity. Telomerase activity is negative in human normal somatic cells but can be detected in most tumor cells. The enzyme is proposed to be an essential factor in cell immortalization and cancer progression. In this review we discuss the structure and function of telomere and telomerase and their roles in cell immortalization and oncogenesis. Simultaneously the experimental studies of telomerase assays for cancer detection and diagnosis are reviewed. Finally, we discuss the potential use of inhibitors of telomerase in anti-cancer therapy. PMID- 11942407 TI - Structure and function of aggrecan. AB - Aggrecan is the major proteoglycan in the articular cartilage. This molecule is important in the proper functioning of articular cartilage because it provides a hydrated gel structure (via its interaction with hyaluronan and link protein) that endows the cartilage with load-bearing properties. It is also crucial in chondroskeletal morphogenesis during development. Aggrecan is a multimodular molecule expressed by chondrocytes. Its core protein is composed of three globular domains (G1, G2, and G3) and a large extended region (CS) between G2 and G3 for glycosaminoglycan chain attachment. G1 comprises the amino terminus of the core protein. This domain has the same structural motif as link protein. Functionally, the G1 domain interacts with hyaluronan acid and link protein, forming stable ternary complexes in the extracellular matrix. G2 is homologous to the tandem repeats of G1 and of link protein and is involved in product processing. G3 makes up the carboxyl terminus of the core protein. It enhances glycosaminoglycan modification and product secretion. Aggrecan plays an important role in mediating chondrocyte-chondrocyte and chondrocyte-matrix interactions through its ability to bind hyaluronan. PMID- 11942408 TI - Identification of gamma1 subunit of GABA(A) receptor in rat testis. AB - The isoform type of gamma subunits of GABA(A) receptor is a molecular determinant of its pharmacological characteristics. At present, the existence of GABA(A) receptor in mammalian sperm is still a controversy. By using degenerate primers designed according to highly conserved region in all three gamma (gamma1, gamma2 and gamma3) subunits cloned in rat brain, we performed reverse transcription polymerase chain reaction (RT-PCR) to examine the expression pattern of gamma subunits of GABA(A) receptor in rat testis. Only one 370 bp fragment was obtained from RT-PCR in rat testis and sequencing results showed that it represented gamma1 subunit, but not gamma2 or gamma3 subunit. Using the cloned fragment as probe, a 3.8 kb transcript which in size as same as gamma1 subunit in rat brain was detected in rat testis mRNA by performing Northern blot assay. Furthermore, results of in situ hybridization assay confirmed that gamma1 subunit was expressed in round spermatids and spermatozoa, maybe also in secondary spermatocyte. These evidences proved that gamma1 subunit of GABA(A) receptor is exclusively expressed in rat testis and this feature may be the structural basis of the specific function of GABA(A) receptors in sperm acrosome reaction. PMID- 11942409 TI - Identification of differentially expressed genes in human uterine leiomyomas using differential display. AB - In searching of differentially expressed genes in human uterine leiomyomas, differential display was used with twelve pairs of primers to compare human uterine leiomyomas with matched myometrium. False positives were eliminated by reverse Northern analysis. Positives were confirmed by Northern blot analysis. RESULTS: Four of 69 cDNA fragments (3 up-regulated named L1, L2 and L3 and 1 down regulated named M1 in leiomyoma) were confirmed by Northern analysis. Sequence comparison and Northern analysis proved that L1 is exactly the human ribosomal protein S19. It was present ubiquitously in 13 tissues tested but in various levels and even in different size. L1 was highly expressed in parotidean cystadenocarcinoma, pancreatic cancer and breast cancer examined. No mutations have been found in human uterine leiomyomas (n=6). CONCLUSIONS: hRPS19 overexpression might be a universal signal in rapid cell growth tissues. PMID- 11942410 TI - The inhibition of lung cancer cell growth by intracellular immunization with LC-1 ScFv. AB - A monoclonal antibody, LC-1, recognizing lung cancer associated common antigens was obtained in authors' laboratory. Its single chain Fv fragment (ScFv) named LC 1 ScFv was constructed based on recombinant phage displayed techniques. For expression on cell membrane, LC-1 ScFv was cloned into pDisplay vector, which directed the cloned gene to express as cell membrane bound protein. The resulting plasmid was sequenced and then introduced by the lipofectin method into a lung adenocarcinoma cell line SPC-A-1. G418 resistant cells were obtained by G418 selection. After transfection, LC-1 ScFv expression was observed by Western blot analysis and the expression of cognate antigens was down-regulated as shown in ELISA assay. SPC-A-1-pDisplay-ScFv cells grew in vitro at lower speed than the control intact cells and the cells transfected with vacant vector. Flow cytometry analysis detected a substantial increase in G1 phase and decrease in S phase in population of SPC-A-1-pDisplay-ScFv cells compared to SPC-A-1 and SPC-A-1 pDisplay cells. Semi-quantitative RT-PCR analysis showed that c-myc expression was down-regulated in SPC-A-1-pDisplay-ScFv cells. It seems that the antigens recognized by LC-1 may be in some way involved in a growth stimulating pathway and the antibody blocking of the function of the antigens shut down the pathway and thus down-regulate the expression of c-myc and growth of the cells. PMID- 11942411 TI - Effects of cisplatin on telomerase activity and telomere length in BEL-7404 human hepatoma cells. AB - Telomerase activity was inhibited in a dose and time-dependent manner with the treatment of cisplatin for 24, 48, or 72 h in a concentration ranged from 0.8 to 50 microM in BEL-7404 human hepatoma cells. There were no changes in expression pattern of three telomerase subunits, its catalytic reverse transcriptase subunit (hTERT), its RNA component (hTR) or the associated protein subunit (TP1), after cisplatin treated for 72 h with indicated concentrations. Mean telomere lengths were decreased by the cisplatin treatment. Cell growth inhibition and cell cycle accumulation in G2/M phase were found to be correlated with telomerase inhibition in the present study, but percentages of cell apoptosis did not change markedly during the process. PMID- 11942412 TI - The molecular characterization of maize B chromosome specific AFLPs. AB - The origin and evolution of B chromosomes could be explained by the specific DNA sequence on them. But the specific sequences known were quite limited. To investigate maize B chromosome sqicific DNA sequeces, maize genomes with and without B chromosomes were analyzed by AFLP. Only 5 markers were found specific to genomes with B chromosomes among about 2000 AFLP markers. Southern hybridization and sequence analysis revealed that only the sequence of M8-2D was a B chromosome specific sequence. This sequence contained the telomeric repeat unit AGG PMID- 11942413 TI - Antibodies against the C-terminal peptide of rabbit oviductin inhibit mouse early embryo development to pass 2-cell stage. AB - A full-length rabbit oviductin cDNA(1909bp) was cloned. It consists of a 5'-UTR of 52bp, an open reading frame (ORF) of 1374bp and a 3'-UTR of 483bp and has more than 80% homology with that of other mammal oviductins. N-terminal peptide (NTP) (384 residues) and C-terminal peptide (CTP) (73 residues) of deduced protein precursor has about 80% and 50% identity with that of other mammals respectively. Fusion proteins GST-NTP 368(1R-368N)and GST-CTP73 (369F-441A) were expressed and purified. NH2-terminal of CTP sequencing reveals that the purified protein is consistent with the deduced one. In order to study the function of NTP and CTP the mouse anti-NTP and rabbit anti-CTP antisera were prepared. Tissue-specific (skeleton muscle, oviduct, uterus, ovary, liver, heart and brain) analysis indicated that rabbit oviductin was only found in oviduct. The conditioned medium derived from the rabbit oviduct mucosa epithelial cells has a function of overcoming the early embryonic development block of Kunming mouse cultured in vitro. Anti-CTP antiserum could totally inhibit the early embryo development at 2 cell stage cultured in the conditioned culture medium, but anti-NTP antiserum couldn't. There was a positive relationship between the ratio of early embryos at development block and the dosage of anti-CTP antiserum added in the conditioned culture medium. These results suggest that oviductin has a function not only on fertilization, but also on the release of early embryonic development block, and the later function domain of rabbit oviductin may be situate in its C-terminal. PMID- 11942414 TI - Identification of a NF-kappaB site in the negative regulatory element (epsilon NRAII) of human epsilon-globin gene and its binding protein NF-kappaB p50 in the nuclei of K562 cells. AB - The developmental control of the human epsilon-globin gene expression is mediated by transcription regulatory elements in the 5' flanking DNA of this gene. Sequence analysis has revealed a DNA motif (GGGGAATTTGCT) similar to NF-kappaB consensus sequence resides in the negative regulatory element (-3028bp approximately -2902bp, termed e-NRAII) 5' to the cap site of this gene. NRF DNA fragment (-3010bp approximately -2986bp) containing the NF-kappaB motif similar sequence was synthesized and used in electrophoresis mobility shift assay (EMSA) and competitive analysis. Data showed that a protein factor from nuclear extracts of K562 cells specifically interacted with NRF DNA fragment. The synthetic NF DNA fragment (containing NF-kappaB consensus sequence) could competed for the protein binding, but MNF DNA fragment (mutated NF-kappaB motif) could not, suggesting that the binding protein is a member of NF-kappaB/Rel family. Western blot assay demonstrated that the molecular weight of NF-kappaB protein in the nuclei of K562 cells is 50ku. We suggested that NF-kappaB p50 may play an important role in the regulation of human epsilon-globin gene expression. PMID- 11942415 TI - MAPK signal pathways in the regulation of cell proliferation in mammalian cells. AB - MAPK families play an important role in complex cellular programs like proliferation, differentiation, development, transformation, and apoptosis. At least three MAPK families have been characterized: extracellular signal-regulated kinase (ERK), Jun kinase (JNK/SAPK) and p38 MAPK. The above effects are fulfilled by regulation of cell cycle engine and other cell proliferation related proteins. In this paper we discussed their functions and cooperation with other signal pathways in regulation of cell proliferation. PMID- 11942416 TI - Evaluating community-based health initiatives: identifying the characteristics of successful initiatives and evaluations. PMID- 11942417 TI - A funny thing happened on the way to managed competition. PMID- 11942418 TI - The Fortune 500 model for health care: is now the time to change? PMID- 11942419 TI - Evaluating partnerships for community health improvement: tracking the footprints. AB - Private-public partnerships are increasingly seen as an important mechanism for improving community health. Despite their popularity, traditional evaluations of these efforts have produced negative or mixed results. This is often attributed to weak interventions or an insufficient period of time to observe an impact. This study examines two additional possibilities--the need for a well-articulated shared vision and the governance and management capabilities of the partnership itself. We conducted a midstream process evaluation of twenty-five community partnerships associated with the Community Care Network (CCN) Demonstration Program. We examined how the roles of a common shared vision, strong governance, and effective management influence a partnership's ability to achieve its objectives. The findings, based on both qualitative and quantitative analyses, underscore the importance of membership organizations' perceived benefits and costs of participation and management capabilities to the partnership's progress toward a vision. Based on the qualitative data, six key governance and management characteristics are identified that separate the top performing partnerships from the lowest performing ones. We explore the implications of this research for future evaluations of public-private community health partnerships. PMID- 11942420 TI - Managed competition versus industrial purchasing of health care among the Fortune 500. AB - The theory of managed competition has found favor with many health policy analysts and academic economists alike. Three characteristics--consumer choice, defined contribution, and dissemination of information--signal managed competition strategy. By requiring private employers to provide their employees with a choice of health carriers, a fixed-dollar strategy (defined contribution), and quality information to make appropriate choices among carriers, managed competition offers to remedy imperfections in both the consumer and provider sides of the market for health insurance. In an extensive survey of health care purchasing practices among Fortune 500 companies we found that major companies are not using the managed competition approach to health care purchasing. Instead, most of the companies surveyed are purchasing health care in the same way as they do other inputs to production--a pattern we call industrial purchasing. PMID- 11942421 TI - Evaluating community partnerships: a response. PMID- 11942422 TI - Poor sleep and psychiatric symptoms at school: an epidemiological study. AB - The objective of this study was to evaluate associations between sleep problems and psychiatric symptoms at school. A random sample consisting of 5813 eight- to nine-year-old children was selected from ordinary schools. Both parents' and children's reports of sleep problems were taken into account. The psychiatric symptoms were addressed according to the teachers' reports (the Rutter Scale B). Children with severe sleep problems were more likely to have a psychiatric disturbance according to the Rutter B Scale (OR 2.45, 95% CI 1.85-3.25). Logistic regression models showed that severe sleep problems were highly associated with emotional problems (OR 2.74, 95% CI 1.84-4.13), school attendance problems (OR 2.53, 95% OR 1.45-4.41), behavioural problems (OR 2.44, 95% CI 1.59-3.75) and hyperactivity (OR 2.02, 95% CI 1.30-3.13). Over 95% of severe sleep problems were reported only by the children themselves. In conclusion, children with severe sleep problems have substantially more teacher-reported psychiatric symptoms than those with no or mild sleep complaints. In diagnosing sleep disorders, it is important to include children as informants because relevant information may be overlooked when only parents are questioned. PMID- 11942423 TI - Early-onset Tourette syndrome with reversible autistic behaviour: a dysmaturational disorder. AB - Early-onset Tourette syndrome comorbid with reversible autistic behaviour is described in twelve young males. After a normal gestation, delivery and first year development, regression set in between the age of one and two with loss of various abilities and the emergence of autistic behaviour. At this time, or slightly later, they showed multiple motor and vocal tics, simple and complex: the latter could also be traced to most of their parents. Following an intervention based on intense cuddling, motor activation and paedagogic guidance, these children's abilities rapidly improved, reaching at follow-up a normal or borderline intellectual functioning and with the disappearance of their initial autistic behaviour. At follow-up tics were present in all, usually with the features of a full-blown Tourette syndrome, often comorbid with ADHD, and in some cases with OCD. PMID- 11942424 TI - DSM-IV conduct disorder symptoms in adolescents as markers of registered criminality. AB - A nationwide sample of 1,079 Norwegian adolescent psychiatric in-patients, 581 males and 498 females, were followed up after 11-33 (mean 23.2) years. On the basis of detailed records from index hospitalisation they were re-diagnosed and retrospectively scored on 14 of the 15 behaviour items listed as diagnostic criteria for conduct disorder in DSM-IV. The patient list was linked to the national criminal registry, and 48% were found to have a criminal record. The ability of the 14 behaviour items to identify overall and specific kinds of registered criminality was investigated. In males, "stealing" was the strongest marker for overall and violent crime. "Having fored someone into sexual activity" was a strong marker for later sex offences. In females, "running away from home" was strongly associated with overall and violent registered criminality. ROC curves were constructed to test the sensitivity and specificity of the sum of conduct disorder behaviour items fulfilled in the identification of individuals with registered criminality. The area under the curve was 0.81 (95% c. i. = 0.78 0.85) in males and 0.75 (95% c. i. = 0.71-0.80) in females. A cutoff at 3 criteria fulfilled seemed optimal in males and somewhat high in females. PMID- 11942425 TI - Attention deficit hyperactivity disorder, combined type (ADHD-CT): differences in blood pressure (BP) due to posture and the child report of anxiety. AB - AIM: ADHD-CT and anxiety has been associated with increased diastolic BP in controlled trials of single dosage studies of methylphenidate, although parent and child reports of anxiety have not been considered separately. Therefore, we tested the hypothesis that primary school-age children with ADHD-CT and anxiety have increased diastolic BP from a sitting to a standing position compared to children with ADHD-CT without anxiety, whether defined from a parent or child perspective. METHOD: Thirty-eight medication naive children with ADHD-CT were studied. Four groups were formed from parent and child reports of anxiety, using categorical and continuous measures of anxiety. The groups were compared using one-way ANOVA. Where omnibus F was significant, the post hoc SNK procedure (p < 0.05) was used to determine the source of this significance. RESULTS: Primary school-age children with ADHD-CT and anxiety, defined from the child's report alone, had significantly increased diastolic BP from a sitting to a standing position than children with ADHD-CT without anxiety. CONCLUSIONS: The recognition of anxiety and its management in primary school-age children with ADHD-CT is generally poorly understood. In this particular group of children, only the child's report of anxiety was associated with significantly increased diastolic BP from a sitting to a standing position. Therefore, careful and thorough assessment of the child's perspective is required. The characteristics of this anxiety and its association with postural BP require further careful longitudinal study. Biological implications are noted. PMID- 11942426 TI - The Youth Self-Report (YSR) and the Depression Self-Rating Scale (DSRS) as measures of depression and suicidality among adolescents. AB - Two hundred and thirty-seven adolescents from a junior high school in a small community outside Goteborg, Sweden, completed the Youth Self Report (YSR) and the Depression Self Rating Scale (DSRS). Self-reported suicidality and biographical data were also recorded. The school doctor and nurse assessed the adolescents' somatic, psychological and behavioural problems using school health-records. The convergent validity of the YSR total problems scale and syndrome scales were tested against the DSRS. Discriminant validity was assessed by the two measures' ability to predict suicidality and school health problems. The Internalising (r = 0.65**) and Anxious/Depressed (r= 0.61**) syndrome scales of the YSR had the highest correlations with the DSRS. However, all YSR syndrome scales were significantly, though more modestly, correlated with the DSRS. Using stepwise logistic regression analysis, four YSR sub-scales [Social Withdrawal, Anxious/Depressed, Attention problems and Delinquency] predicted mild-severe self reported depression (DSRS scores 12 and above). The YSR syndrome scales Anxious/Depressed and Delinquency predicted suicide ideation whereas the Self destructive/Identity problem and Social Withdrawal (low scores) scales predicted Suicide attempts. The YSR Anxious/Depressed sub-scale and the DSRS total score seem to measure a similar dimension. However, the Anxious/Depressed and Selfdestructive/Identity problem scales were superior in predicting suicidality. PMID- 11942427 TI - Do adolescent anorexia nervosa patients have deficits in emotional functioning? AB - Adult eating disorder patients have been characterised by alexithymia. We investigated whether adolescent eating disorder patients also show deficits in emotional functioning. To measure emotional functioning a questionnaire (the TAS) and an emotion recognition test were administered to 30 eating disorder (ED) adolescent girls and 31 healthy controls (HC), matched for age, education, and social status. Non-emotional, cognitive parallel tasks were administered on the same occasion to find out whether a possible deficit was emotion-specific or of a more general cognitive nature. The ED patients scored higher on the TAS and performed worse on the emotion recognition test, but no differences between the groups were found on the non-emotional cognitive instruments. It was concluded that adolescent eating disorder patients, just like adult eating disorder patients, are characterised by alexithymia and show specific deficits in emotional functioning. The implications of these findings are discussed. PMID- 11942428 TI - Neuropsychiatric aspects of Sotos syndrome. A review and two case illustrations. AB - Sotos syndrome, also known as cerebral gigantism, is a rare growth disorder first described by Sotos and colleagues in 1964. Since then, a majority of the literature pertaining to Sotos syndrome has focussed on the medical aspects of the syndrome, while neuropsychiatric issues have been given little attention. The present article presents a review of the current knowledge concerning neuropsychiatric aspects of Sotos syndrome. Information is provided relating to physical appearance, intellectual impairment, learning disabilities, language impairments, behavioural disorders and outcome. It is emphasised that careful delineation of neuropsychiatric aspects within a diagnostic syndrome may turn out to be the first step towards molecular characterisation of behaviour. PMID- 11942429 TI - How NP preceptors can comply with Medicare requirements. PMID- 11942430 TI - Educating children with asthma. PMID- 11942431 TI - Developing the adult NP's role in home care. PMID- 11942432 TI - Achieve new vision screening objectives. AB - Visual impairment ranks among the top ten disabilities in the United States. Early diagnosis and treatment can prevent permanent blindness in many cases. This article identifies the leading causes of visual impairment, target populations, and screening techniques for primary care. The author also provides key counseling information and valuable resources for clinicians and patients. PMID- 11942433 TI - Adult anemia: determine clinical significance. AB - Severity of adult anemia does not reliably indicate its origin or clinical significance. Causes range from dietary insufficiency to occult bleeding and from chronic disease processes to hemolysis. NPs can identify the different types of anemia with a thorough history, physical examination, and appropriate laboratory testing. PMID- 11942434 TI - Understanding anemia. PMID- 11942435 TI - How do we pass NP autonomy legislation? AB - NP practice autonomy means prescribing medication and treatment based on graduate nursing education and credentials, without physician supervision. NP advancement and optimal professional practice depends on autonomy. NP researchers conducted a study to describe legislative initiatives in states where NPs practice and prescribe autonomously. This article outlines the strategies NPs used to pass autonomy legislation in South Carolina. NPs in other states can use these strategies to pursue autonomy legislation. PMID- 11942436 TI - Include UTI in older adults' differential diagnosis of syncope. PMID- 11942437 TI - Market to the media. PMID- 11942439 TI - Improving the advanced health assessment course. PMID- 11942438 TI - Preventing costly falls in long-term care. PMID- 11942440 TI - Debating the doctorate. PMID- 11942441 TI - Learning from death. PMID- 11942442 TI - Assessing change in chronic pain severity: the chronic pain grade compared with retrospective perceptions. AB - BACKGROUND: There is no standard method of measuring change in chronic pain severity. Clinical trials commonly use serial assessment scales, completed at two points in time, to estimate change in pain severity, while clinicians usually ask patients to make a retrospective assessment of change. How the two methods compare is not known. AIM: To assess different methods of measuring change in chronic pain severity, by comparing changes in scores on a serial measure of chronic pain severity using the Chronic Pain Grade (CPG) questionnaire and responders' retrospective perception of change in pain severity. DESIGN OF STUDY: Postal self-completion questionnaires. SETTING: The Grampian region of Scotland. METHOD: Postal questionnaires were sent in March and September 1998 to a random sample of 535 adults with chronic pain, drawn from responders to a postal survey of the region conducted in 1996. RESULTS: Corrected response rates of 87.5% and 90.7% were obtained. Over a six-month period poor levels of agreement were found, with responders' retrospective perceptions mirroring recorded changes in 41.8% of individuals (kappa = 0.081). A low partial correlation coefficient between the two measures (-0.209) was also found. Over a two-year period there were again poor levels of agreement, with responders' retrospective perceptions mirronng recorded changes in 35.2% of individuals (kappa = 0.071). A low partial correlation coefficient (-0.401) was again found. CONCLUSION: There was poor agreement and low correlation between two commonly used methods for assessing change in pain severity over time. This finding has important implications for both service practitioners and researchers. PMID- 11942443 TI - Understanding variation for clinical governance: an illustration using the diagnosis and treatment of sore throat. AB - BACKGROUND: The aim of clinical governance is to improve clinical care. An understanding of the information contained in variation is central to any improvement effort. We must distinguish between variation intrinsic to a process (common cause variation) and variation caused by extrinsic factors (special cause variation). The control chart is a method of distinguishing between these two kinds of variation: it is used in industry to effect improvement and may be useful in primary care. AIM: To illustrate the use of control charts to distinguish between common cause and special cause variation and to guide appropriate action. DESIGN OF STUDY: Analysis of diagnostic and treatment decisions for sore throat. SETTING: Single practice in the West Midlands. METHODS: We identified each general practitioner's (GP's) consultations for sore throat over a two-year period. We grouped these into two diagnostic categories (tonsillitis and non-tonsillar throat infection) and two treatment categories (antibiotics and no antibiotics). These data were illustrated graphically as XY control charts. RESULTS: In this practice, a special cause affects one GP's diagnosis--he is less likely to use the term 'tonsillitis'. A special cause also affects his treatment decisions--he is more likely to prescribe antibiotics. Diagnostic and treatment differences between the remaining GPs are consistent with common cause variation. CONCLUSION: In this practice, action to improve the quality of diagnosis and treatment of sore throat shouldfocus on investigating why one practitioner's diagnosis and treatment differs from that of his colleagues. Control chart analysis is valuable because it enables users to obtain practical guidance for action. PMID- 11942444 TI - Patients with new onset haematuria: assessing the discriminant value of clinical information in relation to urological malignancies. AB - BACKGROUND: There is little information available to assist general practitioners (GPs) in deciding which patients with haematuria are likely to have a malignancy. AIM: To derive discriminant functions for specific items or clusters of clinical history information in relation to the categorisation of patients presenting to the 'open access' haematuria clinic in Hull. DESIGN OF STUDY: Recruitment of patients via an 'open-access' haematuria clinic. SETTING: A consecutive series of 363 patients aged between 18 and 80 years who attended the clinic. METHOD: Between February 1999 and October 1999 clinical history information derived from the participating patients was compared with the patients' diagnoses. Diagnoses were established by a combination of cystoscopy and radiological assessments and rechecked against the patient records and the hospital patient administration system two to three months later. RESULTS: A number of individual variables seemed to be particularly helpful in discriminating malignancies. However, when indicants were combined using regression shrinkage techniques, only the following variables were preserved: age, sex, type of haematuria, number of episodes of haematuria, hesitancy, poor urinary stream, smoking history, and history of urinary tract infections. CONCLUSION: It is possible to generate helpful discriminant information to assist GPs in making more appropriate decisions in a difficult area of clinical practice. However, it remains a matter of judgement as to how representative the study population is likely to be compared with all haematuria patients encountered in primary care. We have reasonable confidence in the general applicability of the rules for macroscopic haematuria: however, it seems likely that the prediction rules outlined for microscopic haematuria have their greatest relevance once a patient has been referred by a GP. In developing the work further and testing out the discriminators identified in this study, we propose that a primary care-based project now needs to be undertaken focusing on microscopic haematuria with a particular emphasis on addressing selection biases. In addition, there is a more general need to assess the reliability of all the suggested items of clinical discriminant information. PMID- 11942445 TI - A randomised controlled trial of the effect of educational outreach by community pharmacists on prescribing in UK general practice. AB - BACKGROUND: Educational outreach visits are commonly used to promote changes in prescribing in family practice. However, the effectiveness of outreach visits has not been evaluated across a range of settings. AIM: To estimate the effectiveness of educational outreach visits on United Kingdom (UK) general practice prescribing and to examine the extent to which practice characteristics influenced outcome. DESIGN OF STUDY: Randomised controlled trial. SETTING: General practices in 12 health authorities in England. METHOD: Educational outreach visits were made to practices that received two of four guidelines. Each practice provided data on treatment of patients for all four guidelines for both pre and post-intervention periods. The primary outcome is average effect across all four guidelines. Secondary analyses examined the predictive effect of practice and guideline characteristics. RESULTS: Seventy per cent of practices approached agreed to take part in the intervention. Overall, educational outreach was associated with a significant improvement in prescribing practice (odds ratio [OR] = 1.24 [95% CI = 1.07 to 1.42]), a 5.2% (95% CI = 1.7% to 8.7%) increase in the number of patients treated within the guideline recommendations. Smaller practices (two or fewer full-time equivalent practitioners) responded much more favourably to educational outreach than larger practices. Smaller practices improved their performance in line with the guidelines by 13.5% (95% CI = 6% to 20.9%) attributable to outreach, while larger practices improved by only 1.4% (95% CI = -2.4% to 5.3%, P-value for interaction <0.001). CONCLUSION: In large practices, educational outreach alone is unlikely to achieve worthwhile change. There is good evidence to support the use of educational outreach visits in small practices. PMID- 11942446 TI - An analysis of practice-level mortality data to inform a health needs assessment. AB - BACKGROUND: The utility of practice death registers has been indicated but, in the wake of the recent Harold Shipman case in the United Kingdom, the value of individual practice-level analysis has been questioned. AIM: To assess the value of analysing practice-level mortality data to inform health needs assessment. DESIGN OF STUDY: Comparative analyses of mortality. SETTING: Two large practices, an inner-city study practice, and a reference practice in a medium-sized town. METHOD: All premature deaths (aged one to 74 years) during 1994-1998 at the study practice (n = 170), and reference practice (n = 340), were identified. Cause specific standardised mortality ratios (SMRs) were calculated using national reference data. The proportions of the total number of years of life lost (YLL) up to age 75 years associated with alcoholism, drug dependency, and severe mental illness were calculated and a comparison between practices was made, using standardised proportional mortality methods. RESULTS: Significantly raised SMRs for the study practice were lung cancer (SMR = 234), digestive system diseases (SMR = 362), and injuries and poisonings (SMR = 180). Having standardised for age, there were nearly four times as many YLLs in the study practice population associated with a history of alcoholism, and over three times as many associated with drug dependency, compared with the reference practice. CONCLUSION: Mortality analyses can provide useful insights for informing needs assessment in an individual practice. Small number problems may occur with smaller practice populations, but collation of data at PCG/T level also has potential utility. The study reinforces the argument that practices need to set up and maintain complete and accurate death registers. PMID- 11942447 TI - Supporting South Asian carers and those they care for: the role of the primary health care team. AB - BACKGROUND: Demographic and socioeconomic changes have increased policy interest in informal carers. However, despite the multicultural nature of British society, most research in this field has been in majority communities. AIM: To explore the role of the primary health care team (PHCT) in supporting carers from British South Asian communities. DESIGN OF STUDY: Qualitative study. SETTING: Four South Asian communities in Leicestershire and West Yorkshire. METHODS: Focus groups and in-depth interviews were used to assess male and female carers, supported by a literature review. RESULTS: Failure to recognise carers' needs, gaps in service provision, and communication and language issues compromised carers' ability to care. While some carers were positive about the PHCT role, the main weaknesses concerned poor consultation, PHCT attitudes towards carers, and access to appropriate services. CONCLUSION: South Asian carers' experiences largely parallel those of others, but there are some issues that are distinct, namely, language and communication barriers, culturally inappropriate services, and implicit or explicit racism. The multi-ethnic nature of Great Britain requires that professional practice enhances the ability of minority ethnic communities to provide informal care. The findings underline the important role of the PHCT in ensuring that carers' needs are taken seriously and that appropriate services reach them. PMID- 11942449 TI - The effectiveness of blood tests in detecting secondary osteoporosis or mimicking conditions in postmenopausal women. AB - The National Service Framework for Older People requires a reduction in the number of falls that result in serious injury. Those most at risk need to be identified, investigated in line with the Royal College of Physicians Clinical Guidelines and receive appropriate treatment. This report looks at the results of investigation of postmenopausal women diagnosed as having osteoporosis in primary care by forearm Dexa scanner (DTX200) and questions whether the investigations suggested within the National Service Framework are justifiable. Scans were performed on 699 postmenopausal women aged 54 or over resulting in a new diagnosis of osteoporosis in 173 women. Complete blood tests were performed in 107 of the newly diagnosed patients. Only three of these patients (2.8%) had blood test results that revealed a potential secondary cause. The rates of positive findings are low; further research is needed to see if they are justified in postmenopausal women. PMID- 11942448 TI - Telephone consultations to manage requests for same-day appointments: a randomised controlled trial in two practices. AB - BACKGROUND: General practitioners (GPs) in the United Kingdom have recently begun to adopt the use of telephone consultation during daytime surgery as a means of managing demand, particularly requests for same-day appointments. However, it is not known whether the strategy actually reduces GP workload. AIM: To investigate how the use of telephone consultations impacts on the management of requests for same-day appointments, on resource use, indicators of clinical care, and patient perceptions of consultations. DESIGN OF STUDY: Randomised controlled trial. SETTING: All patients (n = 388) seeking same-day appointments in each surgery in two urban practices (total population = 10,420) over a four-week period. METHOD: The primary outcome measure was use of doctor time for the index telephone or face-to-face consultation. Secondary outcomes were subsequent use of investigations and of services in the two-week period following consultation, frequency of blood pressure measurement and antibiotic prescriptions, and number of problems considered at consultation. Patient perceptions were measured by the Patient Enablement Instrument (PEI) and reported willingness to use telephone consultations in the future. RESULTS: Telephone consultations took less time (8.2 minutes versus 6.7 minutes; diff = 1.5, 95% confidence interval [CI] = 0.6 to 2.4, P = 0.002). Patients consulting by telephone reconsulted the GP more frequently in the two weeks that followed (0.6 consultations versus 0.4 consultations; diff = 0.2, 95% CI = 0.0 to 0.3, P = 0.01). Blood pressure was measured more often in the group of patients managed face-to-face (25/188 [13.3%] versus 12/181 [6.6%]; diff = 6.7%, 95% CI = 0.6% to 12.7%). There was no significant difference in patient perceptions or other secondary outcomes. CONCLUSION: Use of telephone consultations for same-day appointments was associated with time saving, and did not result in lower PEI scores. Possibly, however, this short-term saving was offset by higher re-consultation and less use of opportunistic health promotion. PMID- 11942450 TI - Mothers' use of and attitudes to BabyCheck. AB - A copy of BabyCheck was sent to 497 mothers shortly after the birth of their baby. Six months later they were sent a questionnaire asking about their use of and attitudes to, BabyCheck. Questionnaires were returned by 323 (65%) mothers; 215 (67%) of them reported reading BabyCheck, the majority found it easy to understand (74%) and agreed with the advice (67%). Eighty-four (26%) of the mothers who returned the questionnaires reported using BabyCheck when their baby was ill; of these, 71% agreed with the advice and 65% trusted the advice. None of the mothers had used the complete range of tests. This lead to our conclusion that BabyCheck is well accepted among mothers but is not used routinely as part of mothers' response to illness in their babies. BabyCheck requires introductory education to ensure that parents score all the tests and that the predictive power of BabyCheck is not compromised. Further evaluation of BabyCheck is needed to find its ideal role in the assessment and monitoring of infant illness. PMID- 11942451 TI - Patient, hospital, and general practitioner characteristics associated with non attendance: a cohort study. AB - Studies examining characteristics of non-attendance at hospital outpatients have given inconsistent results. We examined a cohort of 1972 referrals from 26 general practitioners, with complete follow-up. Five factors were found to be significantly associated with non-attendance: male sex, younger age, longer interval between referral and appointment, higher Jarman score and patients of a high-referring general practitioner. Targeting of strategies to reduce non attendance is possible using these results. PMID- 11942452 TI - Ruptured abdominal aortic aneurysm presenting as groin pain. PMID- 11942454 TI - Systematic review of Viagra RCTs. PMID- 11942453 TI - Clinical governance in Scotland: an educational model. AB - The concepts underpinning clinical governance are similar throughout the United Kingdom but models for its implementation will differ widely. This model aims to enable practices to identify areas for further learning and development against specific outcomes. Criteria sets and standards are suggested and a governance plan is used to allow practices to prioritise their objectives. Resourcing will always be a major issue and such a model should be fully evaluated. PMID- 11942455 TI - Improving the outcome in colorectal cases. PMID- 11942456 TI - Acute assessment of infants presenting to primary care. PMID- 11942457 TI - Reducing benzodiazepine prescribing. PMID- 11942459 TI - Lord of the rings. PMID- 11942460 TI - DTCA in New Zealand: the challenge of finding an acceptable balance. PMID- 11942461 TI - Homelessness and health: Leeds conference. PMID- 11942462 TI - Time, a narrative organiser of events and experience. PMID- 11942464 TI - Dress. PMID- 11942466 TI - The impact of urinary incontinence in working women: a study in a production facility. AB - OBJECTIVES: The objectives of this study were to identify the impact of self reported UI on working women, to describe urine loss symptoms, strategies used to control urine loss, and help-seeking behavior among full-time women working in a rural production facility. METHODS: A cross-sectional survey design was used and questionnaires were distributed to 500 women (response rate, 54%). Items elicited information on demographics, health, parity, symptoms and duration of urine loss, strategies to manage urine loss, effects of UI on work activities, level of knowledge about UI and treatment options, perceived importance of getting professional help, and actual help seeking behavior. RESULTS: Twenty-nine percent (n = 78) reported UI at least monthly. Incontinent women were older (44.8 years vs. 38.1 years) than continent women (t = -5.22, p < .001) and incontinent women had a significantly higher average body mass index (BMI) than continent women (t = -4.3, p < .001). More women reported urine loss with coughing, lifting, bending, and when hands were in water, and were more likely to use pads at work to control urine loss. Few women had reported UI to a health care provider (36%) and most wanted more information about UI (85%). CONCLUSIONS: UI is a prevalent problem for working women. Workplace programs designed to assist women with prevention, treatment, and management of UI are crucial. PMID- 11942467 TI - Coping and HIV infection in African-American women. AB - HIV is impacting African-American women at alarming rates. Many of these women are poor and socially disadvantaged, resulting in a combination of stressors that impacts the quality of their lives. This study investigated whether coping style (i.e., problem-focused, emotion-focused) varies as a function of HIV status or stage of HIV-related illness. Secondly, we examined whether the use of a particular style is associated with three areas of functioning among HIV-infected women: general psychological distress, depressive symptomatology, and physical symptomatology. Ninety-nine HIV-infected women and 143 noninfected women completed measures assessing coping styles and functioning. No significant differences emerged in coping styles between the HIV-infected and noninfected women or for the groups when symptomatic women were examined separate from asymptomatic women. Greater emotion-focused coping was associated with less general psychological distress and depression specifically. Problem-focused coping interacted with illness stage to predict all areas of functioning. By identifying effective coping strategies among African-American women with HIV, mental health professionals can design empirical interventions that can help improve quality of life for these women. PMID- 11942469 TI - Immediate and delayed treatment seeking among adult sexual assault victims. AB - There is a growing body of literature which seeks to better understand the needs of sexual assault victims presenting for specialized treatment. This study explored aspects of immediate and delayed treatment seeking among 1118 women who presented for treatment to a specialized sexual assault care centre within a large urban hospital. Variables related to demographic and assault-specific characteristics were examined for association with immediate (within 12 hours) or delayed (after 12 hours) treatment seeking. Results indicate the severity of the attack prompted women to seek treatment earlier and that women who were assaulted by a known perpetrator were more likely to delay seeking assistance. Findings are conceptualized under the rubric of sociological and feminist frameworks with suggestions for additional research. PMID- 11942468 TI - Presenting the female condom to men: a dyadic analysis of effect of the woman's approach. AB - Although male partner resistance to female condom use has been reported, little is understood about circumstances under which partners will agree to female condom use. This study documents the experiences of couples who have worked together to achieve female condom use. As part of a prospective female condom efficacy study, female participants (age 18-34) received a behavioral intervention and an assortment of take-home items. Selected women and their partners were recruited for a qualitative interview focusing on their experience with the female condom. Interviews were transcribed, double-coded, and verified using a standard retrieval coding system. Twenty-six pairs of linked interviews were analyzed dyadically: 9 couples who used the female condom "consistently," 12 "experimenters," and 5 "non-users." Women who successfully promoted the female condom to their partners used multiple presentation strategies. Initial male partner reaction did not predict continued use beyond the first trial. In conclusion, employment of multiple strategies facilitates successful introduction of the female condom into a sexual partnership. PMID- 11942470 TI - Health promotion in active-duty military women with children. AB - The purpose of this study was to determine the extent to which selected demographic characteristics, definition of health, perceived health status, perceived self-efficacy, and resources are related to the health promoting behaviors of active-duty women with children and to describe qualitatively the experience of being an active-duty mother. Grounded in Pender's (1996) Health Promotion Model, this study used methodological triangulation to test a hypothesized model. A sample of 141 active-duty women with children using military health services participated. Resource availability and commitment were key components of being successful at balancing home and work demands. PMID- 11942471 TI - Prediction of coronary heart disease preventive behaviors in women: a test of the health belief model. AB - Coronary heart disease (CHD) risk increases with age, and this increase is more dramatic in women than men. The incidence of CHD is lower in premenopausal women compared with men. After menopause, the risk of mortality from CHD increases in women. The purpose of this study was to test predictors of CHD preventive behaviors using an adapted form of the Health Belief Model (HBM) in a sample of 178 women. The predictor variables were perceptions of susceptibility to CHD, perceptions of seriousness of CHD, general health motivation, social support, and knowledge of risk factors of CHD. Regression results revealed that susceptibility to CHD, seriousness of CHD, knowledge of risk factors of CHD, and general health motivation together explained 76% of the variance of CHD behaviors. Implications for future research and for improving education about consistent adoption of CHD preventive behaviors are discussed. PMID- 11942472 TI - Cardiac and vascular structure and function are related to lipid peroxidation and metabolism. AB - The present study investigated possible relationships between left ventricular mass, intima-media thickness of the carotid artery (IMT), total arterial compliance, and lipid status in a population sample of 58 apparently healthy subjects aged 20 to 69. By stepwise multiple regression analysis, including age, blood pressure, and smoking, left ventricular mass index, measured by M-mode echocardiography, increased by 13.0 g/m2 for each 1 standard deviation (SD = 0.11 microM, r = 0.60, P< 0.01) increase in plasma malondialdehyde and 9.50 g/m2 per SD increase in plasma 8-iso-prostaglandin F2alpha in women only (SD = 8.88 ng/L, r = 0.44, P = 0.01). Each 1-SD (SD = 0.27 g/L) increase in apolipoprotein B was associated with a 63 microm increase in IMT (r = 0.47, P = 0.014) and a 0.27 mL/min/m2/mm Hg (r = -0.60, P < 0.01) decrease in stroke index/pulse pressure ratio, reflecting total arterial compliance in women. In men, each 1-SD increase in the proportion of stearic acid (18:0) in serum cholesterol esters (SD = 0.12 percent units) reduced the transmitral E/A ratio, measured by Doppler echocardiography, reflecting left ventricular diastolic function, by 0.10 units (r = -0.29, P < 0.05). Thus, important cardiovascular characteristics, such as left ventricular mass, left ventricular diastolic function, carotid IMT, and total arterial compliance, were independently predicted by indices of lipid metabolism and peroxidation in apparently healthy subjects. PMID- 11942473 TI - Fluctuations in human milk long-chain PUFA levels in relation to dietary fish intake. AB - Within the Danish population, milk DHA (22:6n-3) levels vary by more than a factor of 10. This paper deals with fluctuations in the milk content of 22:6n-3 and other long-chain PUFA (LCPUFA) and the acute effects of fish meals and fish oil supplements on milk levels of LCPUFA. Twelve fish-eating mothers with 4-mon old infants provided one blood and one adipose tissue sample, and seven consecutive morning hind-milk samples with dietary records from the previous days. Another 12 lactating women were given fish oil (2-8 g) for breakfast and delivered 6-12 milk samples during the following 24 h. The mean milk 22:6n-3 content of the fish-eating mothers was 0.57+/-0.28 FA% (= percentage of total area of FAME peaks in GLC) and the day-to-day variation (SD/mean) within the individual was 35+/-17%. Mean milk 22:6n-3 content on mornings with no fish the day before was 0.42+/-0.15 FA%; this was increased by 82+/-17% (n = 9, P = 0.05) if the mother had eaten fatty fish. Fish oil resulted in a twofold increase in milk 22:6n-3 levels, which peaked after 10 h and lasted for 24 h. The EPA content of milk was also increased by fish meals and fish oil supplements, but these had no effect on the level of arachidonic acid. The study showed that diurnal and day to-day fluctuations in levels of milk n-3 LCPUFA are large, which makes it difficult to assess the 22:6n-3 intake of breast-fed infants from a single milk sample. In studies of the functional outcome of dietary 22:6n-3 in breast-fed infants it is suggested also to use a measure of maternal 22:6n-3 status. PMID- 11942474 TI - Postprandial and short-term effects of dietary virgin olive oil on oxidant/antioxidant status. AB - It is generally believed that virgin olive oil consumption has beneficial effects, but little is known about its effects postprandially on oxidant/antioxidant status. The aim of this study was to determine changes in oxidative stress biomarkers and lipid profile after a single dose of virgin olive oil and after 1 wk of daily consumption. Sixteen subjects (9 men, 7 women) ingested 50 mL of virgin olive oil in a single dose. Blood samples were collected from 0 to 24 h. Thereafter, 14 participants (8 men, 6 women) followed a 1-wk 25 mg/d virgin olive oil dietary intervention. Blood samples were collected at the end of this period. Serum TAG (P = 0.016), plasma FA (P < 0.001), and lipid peroxidation products in plasma (P< 0.001) and VLDL (P = 0.007) increased, reaching a peak at 4-6 h, and returning to baseline values at 24 h after oil ingestion. The opposite changes were observed in plasma glutathione peroxidase (P = 0.001) and glutathione reductase (GR) (P = 0.042). No changes in LDL lipid peroxidation or resistance to oxidation were observed postprandially. At 24 h, plasma oleic acid remained increased (P < 0.05) and resistance of LDL to oxidation improved (P < 0.05). After 1 wk of virgin olive oil consumption, plasma oleic acid (P = 0.031), resistance of LDL to oxidation (P< 0.05), and plasma GR activity (P = 0.005) increased. These results indicate that changes in oxidant/antioxidant status occur after oral virgin olive oil. Virgin olive oil consumption could provide short-term benefits for LDL resistance to oxidation and in glutathione-related enzyme activities. PMID- 11942475 TI - Dietary n-3 FA modulate long and very long chain FA content, rhodopsin content, and rhodopsin phosphorylation in rat rod outer segment after light exposure. AB - A previous study has shown that the long and very long chain FA (VLCFA) content of the rat retina responds to changes in dietary n-6/n-3 ratio of the fat fed (1). The present study tested whether similar changes in these FA are associated with alterations in rhodopsin content and rhodopsin phosphorylation after light treatment. Weanling rats were fed diets containing 20% (w/w, 40% energy) fat with either high (4.8%, w/w) or low (1.2%, w/w) n-3 FA. After 6 wk of feeding, half of the animals in each group were exposed to light for 48 h at 350 lx or were kept in complete darkness. In the rod outer segment, the high n-3 diet treatment increased the level of 20:5n-3 and 22:6n-3 and reduced the levels of 20:4n-6 and 24:4n-6 in PC, PE, and PS. After the feeding of a high n-3 FA diet, total n-3 pentaenoic VLCFA from C24 to C34 increased in PC, whereas the n-6 tetra- and pentaenoic VLCFA decreased. No changes occurred in n-3 hexaenoic VLCFA regardless of the level of 22:6n-3 in the diet. After light exposure, animals fed a high n-3 FA diet showed reduction in 22:6n-3 as well as in n-6 and n-3 VLCFA in PC. FFA and TG fractions contained increased levels of both 20:4n-6 and 22:6n-3 after light exposure. Dark-adapted rhodopsin content and rhodopsin phosphorylation in the rod outer segment of rats fed the low n-3 FA diet were higher than in animals fed a high n-3 FA diet. After light exposure, animals fed the low n-3 FA diet lost more rhodopsin compared to animals fed the high n-3 FA diet, resulting in less phosphorylation of rhodopsin. Results indicate that the FA composition, rhodopsin content, and phosphorylation in visual cells is influenced by the dietary n-3 FA fed as well as by light exposure. The results also imply that 22:6n-3 may not be the precursor for synthesis of hexaenoic VLCFA. PMID- 11942476 TI - Hypolipidemic action of the soybean isoflavones genistein and genistin in glomerulonephritic rats. AB - Effects of genistein and its glycoside genistin were studied in nephritic rats with endogenous hyperlipidemia. Male Wistar rats with glomerulonephritis caused by a single intravenous injection of nephrotoxic serum were orally given 5 mg of genistein or 8 mg of genistin/d/100 g body weight for 12 d. These isoflavones suppressed nephritis-induced severe hypercholesterolemia and hypertriglyceridemia, and their hypolipidemic action was almost identical. Fecal steroid excretion was unchanged by administration of the two isoflavones. Genistein inhibited the incorporation of [1-14C]acetate into cholesterol and FA in liver slices from nephritic rats when added to an incubation buffer, whereas genistin did not. These results suggest that genistin may be hydrolyzed to genistein and that genistein itself and/or its metabolite(s) may be intracorporal entities suppressing hepatic lipid syntheses. They also suggest that the suppression of hepatic lipid synthesis may be one mechanism of the hypolipidemic action of genistein. PMID- 11942477 TI - Egg ovomucin attenuates hypercholesterolemia in rats and inhibits cholesterol absorption in Caco-2 cells. AB - This experiment was designed to evaluate the effect of casein or ovomucin (OV) on the micellar solubility of cholesterol and the taurocholate binding capacity in vitro. We also evaluated the effects of casein or OV on cholesterol metabolism in rats and Caco-2 cells. OV had a significantly greater bile acid-binding capacity than that of casein in vitro. Micellar cholesterol solubility in vitro was significantly lower in the presence of OV compared to casein. The cholesterol micelles containing OV significantly suppressed cholesterol uptake by Caco-2 cells compared to the cholesterol micelles containing casein. Consistent with these in vitro findings, OV-feeding significantly increased the fecal excretion of bile acids or cholesterol compared with casein-feeding. Serum total cholesterol was significantly lower in rats fed OV than in those fed casein. The concentrations of total lipids in liver were significantly lower in the OV-fed group compared with the casein group. These results suggest that the suppression of cholesterol absorption by direct interaction between cholesterol mixed micelles and OV in the jejunal epithelia is part of the mechanism underlying the hypocholesterolemic action of OV. OV may also inhibit the reabsorption of bile acids in the ileum, thus lowering the serum cholesterol level. PMID- 11942478 TI - Evidence in favor of a facilitated transport system for FA uptake in cultured L6 cells. AB - In this manuscript we report a study of the transport of FA in L6 muscle cells. Cultured L6 cells took up labeled FA (C10 to C20) as a linear function of time up to 15 min. Thereafter, the rate of uptake gradually declined although it persisted for at least 12 h after the addition of the substrate. Kinetic parameters (Km, Vm, and k(o)) were determined from a fitted Michaelis-Menten-type equation modified by a term for a saturable (linear) component of the measured total uptake. Vm values were different for some of the FA studied, and Km data showed significant differences between saturated and unsaturated FA. The maximal rate of uptake was observed at pH 7.40 for decanoate, palmitate, and eicosatrienoate. Uptake was significantly influenced when the pH of the incubation medium was changed. Experiments designed to study the influence of FA/albumin molar ratio indicated that Vm was dependent on the total (bound and free) concentration of the FA. A concentrative uptake was demonstrated in short term experiments with an apparent plateau of 20 and 40 microM for palmitate and eicosatrienoate, respectively. A competitive inhibition was also observed between palmitate as substrate and the other FA. From our results we can postulate that the uptake of FA in L6 cells is the sum of passive diffusion plus a saturable component and that the rate of uptake is dependent on one (or more) protein structures, although their precise characteristics and functions remain to be elucidated. PMID- 11942479 TI - Platelet phospholipids are differentially protected against oxidative degradation by plasmalogens. AB - The oxidative degradation of phospholipids in the presence and absence of plasmalogens (plasmenyl phosphatidylethanolamine: PPE) was followed by chemical analysis. Human platelet phospholipids, either intact or after removal of PPE by acid treatment, were oxidized with 28 mM 2,2'-azobis(2-amidinopropane di-HCl in Triton X-100 micelles (detergent/phospholipid 5:1, mol/mol). PPE (12% of all phospholipids, mol/mol) disappeared about three times more rapidly than glycerophospholipids, whereas sphingomyelin remained unaltered and the lysophosphatidylethanolamine (lysoPE) generated became progressively more unsaturated. After 60 min oxidation, the FA compositions of PS, PC, and PI were similar in extracts with or without plasmalogens. In contrast, diacyl phosphatidylethanolamine (DPE) became more saturated in the absence of PPE. The rate of phospholipid destruction was always unique to each class, but for all phospholipids slowed down in the presence of PPE. This protective effect increased in the order DPE < PS < PC < PI and did not seem to be simply related to the class unsaturation. Alpha-tocopherol had no influence on the time courses of the quantities and compositions of the phospholipids, even at a molar ratio of alpha-tocopherol to phospholipids four times higher than in platelet membranes. Thus, PPE protected phospholipids efficiently but differentially against peroxidative attack, whereas the contribution of alpha-tocopherol appeared to be negligible even at a concentration four times greater than in platelet membranes. PMID- 11942480 TI - Lung lipid composition in zinc-deficient rats. AB - There have been a limited number of studies investigating surfactant lipid changes in lung with trace elements. The present investigation was designed to examine the effect of moderate zinc deficiency on the lipid metabolism in rat lung. We also evaluated whether zinc deficiency, which is a wide-spread problem, could play a role in adult respiratory distress syndrome (ARDS). For that purpose, adult male Wistar rats were fed two diets differing in zinc concentration. The rats were divided into two groups. One group was fed a zinc deficient diet containing 3 mg Zn/kg, and the other group received a zinc adequate control diet with 30 mg Zn/kg according to AIN 93-M. After 2 mon of treatment, we observed that in the zinc-deficient group (i) total lipids, phospholipids, and cholesterol increased whereas TG decreased in whole lung; (ii) phospholipid (PC) concentration increased in lamellar bodies and alveolar macrophages and decreased in extracellular surfactant but did not change in microsomes; (iii) protein concentration decreased in whole lung, extracellular surfactant, lamellar bodies, and macrophages; (iv) the incorporation of [Me 14C]choline into PC (phospholipids) of lung slices increased; and (v) the activity of CTP/phosphocholine cytidylyltransferase bound to the microsomes increased in the lung. These results suggest that the lipid concentration in the lung (especially the phospholipids) is modified directly or indirectly by a zinc deficient diet. In a zinc-deficient diet, the lung changes the pattern of PC for an adaptive or recovery stage. Therefore, zinc deficiency implications are important for the design of therapies and public health interventions involving targeted zinc supplementation for high-risk groups or groups with certain diseases, such as ARDS. PMID- 11942481 TI - Lipid metabolism and FA composition in tissues of Eurasian perch Perca fluviatilis as influenced by dietary fats. AB - Eurasian perch, Perca fluviatilis, were fed a semipurified fat-free diet for 4 wk, followed by a 16% feeding supplementation of either olive oil (OO), safflower oil (SO), linseed oil (LO), or cod liver oil (CLO) as the only lipid source in each diet for 10 wk. Significant reductions in total lipid of tissues were observed (31.4% in viscera, 66.7% in muscle, and 74.1% in liver) after feeding the fat-free diet. The SO-, LO-, and CLO-fed fish significantly increased lipid deposition in liver and viscera compared to fish fed the OO diet; however, muscle lipid levels were not significantly affected. Large amounts of dietary 18:1 n-9 were incorporated directly into tissue lipids when fish were fed the OO diet. The LO diet significantly elevated 18:4n-3, 20:5n-3, 22:5n-3, and 22:6n-3 in the liver compared to fish fed OO or SO diets, and the n-3/n-6 ratio was 16 times that of the SO group, with significantly high desaturation and elongation products of 18:3n-3. These results suggest that delta6 and delta5 desaturases are highly active in Eurasian perch, and that the enzymes at this dietary n-3/n-6 ratio favor 18:3n-3 over 18:2n-6 as substrate. The SO diet significantly increased 18:3n-6, 20:3n-6, and 22:5n-6 in the liver and significantly decreased EPA and DHA. This indicates that desaturation enzymes were not specifically favoring n-3 over n-6 acids in perch lipid metabolism, and that these elongation and desaturation enzymes were influenced by n-3 and n-6 FA content in the diet. The present study indicates that high tissue content of DHA in the muscle of Eurasian perch was attributable to the greater ability for n-3 acid bioconversion. PMID- 11942482 TI - Novel methoxylated FA from the Caribbean sponge Spheciospongia cuspidifera. AB - The delta6 monoenoic methoxylated FA (6Z)-2-methoxy-6-heptadecenoic acid and (6Z) 2-methoxy-6-octadecenoic acid were identified for the first time in nature in the phospholipids from the uncommon Caribbean sponge Spheciospongia cuspidifera. These findings expand the occurrence of A6 2-methoxylated FA to C17-C18 chain lengths and establish a new FA biosynthetic possibility for these marine organisms. The novel methoxylated FA also could have originated from the phospholipids of a bacterium in symbiosis with the sponge. PMID- 11942483 TI - Changes in lipid molecular species and sterols of microsomal membranes during aging of potato (Solanum tuberosum L.) seed-tubers. AB - Changes in sterols and the molecular species composition of polar lipids from microsomal membranes were characterized as a prerequisite to determining how lipid chemistry affects membrane susceptibility to peroxidation during aging of potato tubers. Polar lipid content of the microsomal fraction fell 17% (protein basis) as tubers aged from 22 to 38 mon at 4 degrees C. In younger seed-tubers, PC concentration (protein basis) was the highest, followed by digalactosyldiacylglycerol (DGDG), PE, monogalactosyldiacylglycerol (MGDG), and PI. PC and PE increased 14 and 27%, respectively, whereas glycolipids fell 64 and PI 43% with advancing age. These changes resulted in PC and PE dominating the microsomal membrane lipids of 38-mon-old tubers. Nonpositional analysis of lipid molecular species across lipid pools showed an increase in 16:0/18:3, 18:3/18:3, and 18:2/18:3 (PC and PE only), and a decline in 18:2/18:2 and 16:0/18:2 (except for MGDG) with advancing tuber age. The increase in 18:3-bearing species effected a linear increase in double-bond index (DBI) of PC and PE during aging. The DBI of DGDG did not change with age; however, it fell 65% for MGDG, resulting in an overall decrease in average microsomal DBI. In addition, A5-avenasterol and stigmasterol concentrations increased 1.6- and 3.3-fold, respectively, effecting a significant increase in the sterol/phospholipid ratio with advancing tuber age. The increase in sterol/phospholipid ratio and the possibility that the increased unsaturation of microsomal membranes reflects a compensatory response to maintain optimal membrane function in light of the age-induced loss of galactolipid and PI are discussed. PMID- 11942484 TI - Selective (R)-3-hydroxylation of FA by Stenotrophomonas maltophilia. AB - Soil samples were screened for microorganisms selectively transforming FA. One of the isolated strains was identified as the bacterium Stenotrophomonas maltophilia by its phenotypic features and genotypic characterization by sequencing the ribosomal RNA gene. Using linoleic acid as substrate resulted in the formation of two major compounds. After liquid chromatographic isolation and separation, their structures were elucidated by HPLC-tandem MS, GC-MS, and NMR techniques to be 3 hydroxy-Z6-dodecenoic acid and 3-hydroxy-Z5,Z8-tetradecadienoic acid. In additional experiments, other FA, such as a-linolenic, oleic, palmitoleic, myristoleic, and cis-vaccenic acids, were converted to 3-hydroxylated metabolites of shorter chain lengths as well. Determination of the enantiomeric composition revealed highly enriched (R)-hydroxylation (88-98% enantiomeric excess). PMID- 11942485 TI - Saturated and unsaturated anteiso-C19 acids in the seed lipids from Hesperopeuce mertensiana (Pinaceae). AB - Minor uncommon FA from Hesperopeuce mertensiana (a gymnosperm species of the Pinaceae family) seed oil were characterized through a combination of silver ion TLC of their FAME, and GLC coupled with MS of their picolinyl derivatives. These uncommon components have the structures 16-methyloctadecanoic (anteiso-19:0), 16 methyl-cis-9-octadecenoic (anteiso-19:1), and 1 6-methyl-cis-9,cis-12 octadecadienoic (anteiso-19:2) acids. These branched C19 acids were identified earlier in the wood of Picea abies, which would indicate that such acids could be widespread, though minor, components of Pinaceae lipids. PMID- 11942486 TI - Distribution of medium-chain FA in different lipid classes after administration of specific structured TAG in rats. AB - Structured TAG (STAG) containing medium-chain FA (MCFA) in the sn-1,3 positions and essential FA in the sn-2 position were synthesized by lipase-catalyzed acidolysis. In our previous studies we found that part of the MCFA from STAG could be absorbed in the small intestine; however, it was unclear how they were absorbed. In order to get a better understanding of the metabolism of STAG to improve future design and application of STAG, in the present study lymph lipids collected after feeding STAG were fractionated into different classes and the FA composition of each lipid class was studied by GC after methylation to FAME. Caprylic acid was detected in the fraction of TAG only after administration of 1,3-dioctanoyl-2-linoleyl-sn-glycerol (8:0/18:2/8:0), whereas lauric acid was detected in TAG, DAG, and FFA as well as phospholipids after administration of 1,3-didodecanoyl-2-linoleyl-sn-glycerol (12:0/18:2/12:0). We conclude that the enterocyte has the ability to reacylate the MCFA into TAG and that the intestinal absorption of MCFA from STAG mainly occurs by resynthesis of TAG. Caprylic acid from STAG is not incorporated into phospholipids, whereas lauric acid from STAG can be incorporated into phospholipids. PMID- 11942487 TI - Ruptured uterus: an ongoing tragedy of motherhood. AB - This study aimed to determine the frequency of ruptured uterus, possible aetiologic factors, foetomaternal out come and changes in obstetric care proposed to reduce this catastrophy. From September 1994 to September 1999 in Dhaka Medical College Hospital 39,782 deliveries occurred. All delivery records were reviewed and rupture uterus cases were identified. The result was compared with studies carried out in home and abroad. During the five years period among 39,782 deliveries 424 cases were of rupture uterus with a frequency of 1/93 deliveries. Eighty three percent rupture occurred in intact uterus and 17% occurred in uteri scarred by caesarian section. Common contributing factors were prolonged/obstructed labour, grand multiparity, injudicious use of uterine stimulants, mismanaged labour by traditional birth attendant, delayed referral to well equipped centre, poor communication, poverty and ignorance. PMID- 11942488 TI - Comparison of bone mineral density between premenopausal and postmenopausal women in Bangladesh. AB - This cross sectional study was conducted in the department of Obstetrics and Gynaecology at Bangabandhu Sheikh Mujib Medical University, Dhaka during July 1997 to June 1998. Two hundred and twenty (220) Bangladeshi premenopausal and post-menopausal women aged from 35 to 55 years were studied to compare the effect of age, parity, socioeconomic status and nutritional status on bone mineral density (BMD) of distal and ultra distal ends of radius and ulna. A bone densitometer (Single photon X-ray absorptiometry, DTX100, USA) was used to measure the BMD. BMD was calculated by taking the mean of the two sites and is expressed in g/cm2. Bone mineral density in post-menopausal women (0.48 +/- 0.1 g/cm2) was significantly lower than pre-menopausal women (0.61 +/- 0.06 g/cm2, p<0.001). Parity and number of family members were significantly greater in post menopausal women compared to pre-menopausal women (6.5 +/- 2.9 vs. 4.2 +/- 2.3 and 8.1 +/- 2.5 vs. 6.2 +/- 2.3 respectively, p<0.001). Height was significantly lower in post-menopausal women (148.6 +/- 6.9 vs. 151.8 +/- 5.7 cm, p<0.001). After adjustment of various biological factors in multiple linear regression, body mass index (BMI) showed negative relationship with age (slope -0.0068, p<0.001) and positive relationship with weight (slope 0.0011, p<0.05). Peak BMD was observed upto the age of 40 years and thereafter declined and sharply after menopause. The study concluded that bone mineral density is greater in pre menopausal women than post-menopausal women and remains high up to 35-40 years of age, and declines to older age. BMD positively correlates with weight of women. Premenopausal women have higher height than postmenopausal women though height does not have correlation with BMD in this study. PMID- 11942489 TI - Risk factors for acute respiratory infections among the slum infants of Dhaka city. AB - A cross-sectional study was conducted in five slums of Dhaka city. The study population was all adult women of reproductive age having an infant aged less than one year with a view to assessing the prevalence of Acute Respiratory Infections (ARIs) among the slum infants and also to identify the factors responsible for it. A total of 1,008 mothers were interviewed. Among them, 927 had children aged 0-12 months. Mothers with live infants (927) were asked if their children had any disease during the two weeks prior to the interview. Respiratory infection was the highest among the prevalent diseases (ARI 72%, diarrhoeal diseases 28%, measles 4% and others 4%). So, an emphasis was given in this regard. Out of 228 ARI cases, percentage of mild (78%) and severe ARI (11%) were present in children aged less than 6 months, whereas moderate ARI (23%) was higher in 6 months and above age groups. The variation was statistically significant between the two age groups (p<0.05). But there was no significant difference in ARI by sex (p>0.05). In logistic analysis, maternal age below 20 years, working mothers, low housing and socio-economic index, no household possession, no access to piped water and infant's age above 6 months appeared to be significant predictors of ARIs. The risk of acquiring ARI was 3.33 times higher in low socio-economic index, 3 times in no access to piped water, 2.39 times in low housing index, 1.9 times in mother's age below 20 years, 1.85 times in infant's age above 6 months, 1.69 times in working mothers. On the other hand, household possessions had protective effects on ARIs. The study provides important information for policy makers regarding the prevention of ARI among the children of the slum dwellers. PMID- 11942490 TI - Clinicopathological correlation of acute appendicitis. AB - Acute appendicitis is the most common acute surgical condition of the abdomen and appendicectomy is the most frequently performed urgent abdominal operation. Delay in treatment of acute appendicitis causes lot of complications. On the other hand, to reduce complications prompt diagnosis and treatment results lot of negative laparotomy (as high as 25%). The prime aim of this study was to reduce the incidence of negative laparotomy. PMID- 11942491 TI - Stroke: seasonal variation and association with hypertension. AB - A prospective study was contemplated with 155 patients of acute stroke in the Department of Neurology, Bangabandhu Sheikh Mujib Medical University (BSMMU), Dhaka, from March 1996 to February 1997. The objective of the study was to find out seasonal variation of stroke occurrence. An attempt was also made to find out the association of hypertension with ischaemic and haemorrhagic stroke during summer and winter. The numberofsubjects duringsummer and winterwere 121 and 34, respectively. In summer, out of 121 subjects, 87(71.9%) were ischaemic stroke and 34(28.1%) haemorrhagic stroke, of which 32 were intracerebral haemorrhage (ICH) and 2 subarachnoid haemorrhage (SAH). There were 18 (52.9%) ischaemic stroke and 16(47.06%) haemorrhagic stroke (all ICH) among total 34 subjects during winter. The frequency of ischaemic stroke during summer was significantly greater than that during winter (p<0.05). The frequency of haemorrhagic stroke during winter was significantly greater than that during summer (p<0.05). Temperature was positively correlated to ischaemic stroke (p<0.01) and negatively correlated to haemorrhagic stroke (p<0.1). Hypertension was significantly more associated with haemorrhagic stroke than with ischaemic stroke irrespective of season (p<0.05). PMID- 11942492 TI - Whole-brain functional magnetic resonance imaging of cerebral arteriovenous malformations involving the motor pathways. AB - To investigate cortical, basal ganglia and cerebellar activation in patients with arteriovenous malformations (AVMs) involving the motor pathways, we studied ten patients (six male, four female, mean age 30.3 years, range 7.4-44.1) by whole brain functional magnetic resonance imaging (fMRI) in a 1.5-T scanner with the EPI-BOLD-technique. In seven cases multiple fMRI studies were available, acquired in the course of the multi-session endovascular interventional treatment. Self paced right- and left-handed finger-tapping tasks were used to invoke activation. In six patients a super-selective amytal test (Wada test) was performed during diagnostic pre-interventional angiography studies. Abnormal cortical activation patterns, with activation of the primary sensorimotor area, the supplementary motor area and/or the cerebellum shifted to unphysiological locations, were found in four patients. In all cases, localization of the AVM could account for the changes from the normal. After endovascular procedures, fMRI demonstrated shifts in the activation pattern in three patients. In the six patients that had undergone fMRI studies and the Wada test, both methods yielded comparable results. The fact that AVMs are structural anomalies for which the brain can partly compensate ('plasticity') was underlined by these results. fMRI is a valuable tool in the pre-therapeutic evaluation and post-interventional follow-up of patients with cerebral AVMs in whom an operation or an endovascular procedure is planned. PMID- 11942494 TI - Contrast-enhanced MR angiography (enhanced 3-D fast gradient echo) for diagnosis of cerebral aneurysms. AB - We evaluated contrast-enhanced MRA (enhanced 3-D fast gradient echo, EFGRE3D, with spectral IR) for the identification of unruptured cerebral aneurysms. We examined 22 aneurysms from 15 patients. In its ability to identify aneurysms, contrast-enhanced MRA was superior to 3-D-time of flight (3D-TOF) MRA in 4 instances, equal in 17, and inferior in 1. Contrast-enhanced MRA demonstrated thrombus formation more clearly and accurately than did 3D-TOF MRA. Contrast enhanced MRA was performed in less than 1 min, and was found to be a useful method for the identification of intracranial aneurysms. PMID- 11942493 TI - Spontaneous regression of cerebral arteriovenous malformations: clinical and angiographic analysis with review of the literature. AB - Spontaneous regression of cerebral arteriovenous malformation (AVM) is rare and poorly understood. We reviewed the clinical and angiographic findings in patients who had spontaneous regression of cerebral AVMs to determine whether common features were present. The clinical and angiographic findings of four cases from our series and 29 cases from the literature were retrospectively reviewed. The clinical and angiographic features analyzed were: age at diagnosis, initial presentation, venous drainage pattern, number of draining veins, location of the AVM, number of arterial feeders, clinical events during the interval period to thrombosis, and interval period to spontaneous thrombosis. Common clinical and angiographic features of spontaneous regression of cerebral AVMs are: intracranial hemorrhage as an initial presentation, small AVMs, and a single draining vein. Spontaneous regression of cerebral AVMs can not be predicted by clinical or angiographic features, therefore it should not be considered as an option in cerebral AVM management, despite its proven occurrence. PMID- 11942495 TI - Pooling of echographic contrast agents during transcranial Doppler sonography: a sign in favor of slow-flowing giant saccular aneurysms. AB - Transcranial Doppler (TCD) sonography is a new imaging technique allowing for non invasive evaluation of the intracranial vascular anatomy and cerebral hemodynamics. The recent introduction of echographic contrast agents has significantly increased the sensitivity of TCD for the diagnosis of intracranial vascular lesions. We report a case of giant AComA aneurysm, undetected by color and power TCD, which became visible after echographic contrast administration as a delayed and persistent area of contrast enhancement. Knowledge of atypical or contrast-specific lesion appearances will become important if contrast-enhanced TCD is to be used routinely in the diagnosis of intracranial aneurysms. PMID- 11942496 TI - Extravasation on three-dimensional CT angiography in patients with acute subarachnoid hemorrhage and ruptured aneurysm. AB - Three-dimensional computed tomographic angiography (CTA) is a noninvasive technique for detecting lesions after acute subarachnoid hemorrhage. We encountered extravasation on CTA, a finding that has not been reported previously. Three patients with saccular aneurysms showed extravasation on CTA performed within 3 h of the onset of hemorrhage, and all three patients died within 2 weeks. At autopsy, the site of rupture of the aneurysm was confirmed in all three cases. There were two patterns of extravasation shown by CTA, which seemed to depend on the direction of rupture. Extravasation on CTA might represent the natural progression of ruptured aneurysm and may indicate a poor prognosis. PMID- 11942497 TI - MR detection of microhemorrhages in neurologically healthy adults. AB - T2*-weighted gradient-echo (GE) magnetic resonance images frequently demonstrate small hypointense lesions in patients with systemic hypertension and spontaneous hematomas. These lesions have been suspected to represent subclinical microhemorrhages. We examined the incidence of these lesions in neurologically healthy adults, and the factors associated with them. Axial T2*-weighted GE images (TR = 1,000 ms, TE = 30 ms, flip angle = 20 degrees) were obtained in addition to conventional T1- and T2-weighted spin echo images in 450 neurologically healthy Japanese adults (289 men and 161 women; age 52.9 +/- 7.7 years, range 24-84). The overall incidence of small hypointense lesions was 3.1% (14/450), and these lesions were closely related to systemic hypertension (P < 0.0001) and heavy cigarette smoking (>20 cigarettes per day; P=0.003). Although the incidence of hypointense lesions was lower in neurologically healthy adults than in the reported incidence in patients with a hemorrhagic history, the presence of these lesions was related to the risk factors for primary intracerebral hemorrhage even in the neurologically healthy adults. PMID- 11942498 TI - Proton MR spectroscopic imaging in multiple sclerosis. AB - We studied 24 patients with multiple sclerosis (MS) by proton magnetic resonance spectroscopic imaging (1H-MRSI) to assess the neurochemical pathology of the white-matter lesions (WML) and normal-appearing white matter (NAWM). Our 1H-MRSI technique allowed simultaneous measurement of N-acetylaspartate (NAA), choline containing compounds (Cho), and creatine plus phosphocreatine (Cr) signal intensities from four 15-mm slices divided into 0.84 ml single-volume elements. In WML we found significantly lower NAA/Cr and NAA/Cho ratios and a significantly higher Cho/Cr ratio than in NAWM or control white matter. In NAWM, NAA/Cr and Cho/Cr were significantly lower than in control white matter. 1H-MRSI was compatible with damage to myelin in WML, and with axonal damage and/or dysfunction in WML and NAWM. These findings extend data on involvement of NAWM in MS beyond the abnormalities visible on MRI. PMID- 11942499 TI - Five-year retrospective changes in hippocampal atrophy and cognitive screening test performances in very mild Alzheimer's disease: the Tajiri Project. AB - The medial temporal lobe, especially the hippocampus, is important for normal cognitive function. especially for memory, and is the region with the earliest and most extensive pathological changes in Alzheimer's disease (AD). We investigated the atrophic changes of the hippocampus over a 5-year period and its relation to cognitive screening test performances in normal elderly subjects, those with very mild AD, and patients with AD. Fifty-seven elderly subjects without a moderate or greater degree of cerebrovascular disease as shown by MRI were randomly selected from the town of Tajiri. Thirty-three subjects with a clinical dementia rating (CDR) of 0 (normal), 18 CDR-0.5 (very mild AD) subjects, and six CDR-1&2 (AD) subjects underwent MRI and the Mini Mental State Examination (MMSE) twice during the period. Retrospective changes in the hippocampal width and the MMSE scores were evaluated. There were significant CDR group effects for the changes in the mean bilateral hippocampal widths and the MMSE scores. Normal subjects did not show cognitive decline, although there was a slight tendency for hippocampal atrophy. A significant and meaningful Spearman's correlation was noted between left hippocampal atrophy and the MMSE scores over the 5-year period for the CDR-0.5 group. These CDR-0.5 subjects met the MCI (mild cognitive impairment) criteria as proposed by the consensus paper. Findings suggested that normal elderly subjects maintain a high level of cognitive functions for at least 5 years, although hippocampal atrophy might occur. Atrophic change of the left hippocampus might be a good marker of the very early stage of AD. PMID- 11942500 TI - Unilateral absence of cerebellar hemisphere: a case report. AB - We describe a 38-year-old woman with absence of right cerebellar hemisphere incidentally discovered by MR imaging. No cerebellar abnormality was detected on neurological examination. Tissue probably representing dysgenetic cerebellar tissue with no corticomedullary differentiation was present, connected to the right superior cerebellar peduncle. Ipsilateral enlargement of the pons and cerebral peduncle were additional findings. Although the terms "aplasia" or "agenesis" have been used to describe this entity, intrauterine destruction is the presumed pathogenetic mechanism in our case, and therefore these terms have been avoided. Asymmetry of pons and mesencephalon may be related to compensatory reorganisation or to the impairment of sequential development of nuclei and neural tracts. PMID- 11942501 TI - Magnetic resonance imaging following fat obliteration of the frontal sinus. AB - The paper describes the evaluation of magnetic resonance imaging (MRI) following osteoplastic flap procedure with fat obliteration. MRI scans performed in patients after surgery between 1st January 1986 and 31st December 1997 were evaluated. Outcome parameters were time-dependent changes in the distribution of adipose or connective tissue, development of necroses or oil cysts, recurrences, inflammatory complications, or mucocoeles. Eighty-six postoperative MRI scans from 51 operations were evaluated. In 19 cases between two and five MRI scans were available. Time between surgery and the last MRI scan was 24.1 months on average. We found five mucocoeles. The amount of adipose tissue depictable on the last scan was less than 20% in the majority of cases (53%) and more than 60% in only 18% of cases. Statistical tests and modelling showed a significant decrease of adipose tissue with time, with a median half-life of 15.4 months in a subgroup with at least two MRIs. MRI is at times the most valuable diagnostic tool after frontal sinus obliteration using adipose tissue. The method has some limitations with regard to detection of small (recurrences of) mucocoeles and differentiation between vital adipose tissue and fat necroses in the form of oil cysts. In difficult cases long-term MRI follow-up is necessary for definitive evaluation. PMID- 11942502 TI - Prognostic value of MRI findings in sciatica. AB - The natural course of sciatica due to disc herniation is generally favourable but individually unpredictable. Some patients recover only after prolonged conservative therapy or surgery. This study aims to ascertain whether magnetic resonance (MR) imaging features can be used to predict outcome of sciatica and help to identify patients unlikely to respond to conservative management. For a transversal diagnostic study 274 primary care patients underwent early MR imaging for leg pain. One hundred and thirty-three patients with sciatica were followed for 3 months, both patients and physicians being unaware of MR imaging findings. At 12 weeks a favourable prognosis was indicated by the following features: annular rupture (P= 0.02) and nerve root compression on MR imaging (P = 0.03). Poor prognosis was indicated by disc herniation in the foramen (P = 0.004). Our findings show that early MR imaging features are related to prognosis. However, the associations are not strong enough to justify routine use of early MR imaging to predict the prognosis of sciatica. PMID- 11942503 TI - Early MRI findings in stab wound of the cervical spine: two case reports. AB - MR imaging was found to be the most sensitive modality for the detection of spinal cord abnormalities in the acutely injured spine. Although it is reported that traumatic pneumomyelogram indicates a base-of-skull or middle cranial fossa fracture and is almost certainly associated with intracranial subarachnoid air, early MR imaging may demonstrate subarachnoid air in penetrating trauma of the spinal cord without head injury. We report two cervical-spine stab-wound cases, one of which had subarachnoid air on early MR findings. PMID- 11942504 TI - Stent impact on the geometry of the carotid bifurcation and the course of the internal carotid artery. AB - A measurement system is proposed to evaluate reconstructive effects of carotid stents on the geometry of the carotid bifurcation and the course of the internal carotid artery. To describe deviations of the stenotic internal carotid artery (ICA) from the extended axis of the common carotid artery (CCA) the CCA-ICA angle is measured between the CCA midaxis and the midaxis of the stenotic ICA segment. Maximal extensions of ICA tortuosities perpendicular to the course of the CCA axis are defined as ICA offset. The measurements were applied to DSA images of 224 carotid stenoses to evaluate variation and correlation between the two parameters. Comparative pre- and post-stent evaluation was performed in two series of 55 and 31 carotid stenoses treated with Wallstents and in a historic control group of 35 stenoses treated with Strecker stents. Straight course of the ICA was associated with low angle and low offset values, whereas tortuous course of the ICA showed larger angle and offset. A moderate linear correlation between the two parameters was found. Corresponding to a straightening of the stented segment, Wallstents reduced mean angle and offset values significantly. In five cases of the second series of Wallstents, transferrals of curves above the distal stent end associated with kinks were observed, and offset remained constant or increased. Strecker stent implantation caused no significant changes of bifurcational geometry. The proposed parameters corresponded to visual aspects of ICA tortuosity and detected reconstructive effects of self-expanding Wallstents on the ICA course. The measurement system may provide a basis for geometric evaluation of different stent types or implantation concepts with the aim: to optimize anatomic recanalization results in tortuous high angle-high offset bifurcations. PMID- 11942505 TI - Sequential MRI and MR spectroscopy in embolized meningiomas: correlation with surgical and histopathological findings. AB - The preoperative embolization of meningiomas is commonly used to facilitate surgery. The purpose of this study was to evaluate the morphological and metabolic changes in embolized meningiomas and to correlate the results with surgical and histopathological findings. In a prospective study, 36 patients with intracranial meningiomas were included. The extent of devascularization was assessed by angiography and MR volumetry. MRI and MR spectroscopy (MRS) were performed before and sequentially after embolization. At surgery, blood loss was measured and intraoperative duplex-mode ultrasound was applied to identify avascular tumor portions. Histopathological specimens were evaluated for the histological subtype, localization and extent of necrotic tumor portions. Postembolization MRI revealed a variable pattern of secondary revascularization and devascularization with an early onset following embolization. In all patients, peripheral secondary enhancement was present which histopathologically represented a thin layer of vital tumor tissue. MRS revealed lactate in devascularized areas immediately after embolization. Lipids were not observed before the 3rd day after embolization and were always associated with avascular and soft tissue at the time of surgery. Embolized meningiomas feature a variable dynamic with the potential for revascularization and secondary devascularization. Lipid signals indicate avascular and soft tissue at surgery. In case of delayed surgery, MRI and MRS should be performed in order to exclude revascularization and to establish the fatty degeneration of the meningioma. PMID- 11942506 TI - Evaluation of topography and vascularization of cervical paragangliomas by magnetic resonance imaging and color duplex sonography. AB - The management of cervical paragangliomas (PGs) depends on their specific type and their relation to adjacent vessels. The purpose of this study was to evaluate the potential of magnetic resonance imaging (MRI) and color duplex sonography (CDS) to classify PGs according to topography and vascularization. Sixteen patients harboring 22 PGs were studied retrospectively. With digital subtraction angiography as reference, the topographic relation of the tumors to the carotid arteries and the internal jugular vein and the patterns of vascularization were assessed. On MRI and CDS the typical feature of 15 carotid PGs was splaying of the carotid bifurcation, with the external carotid displaced anteriorly and the internal carotid artery and internal jugular vein located posteriorly. In five vagal PGs both modalities showed unidirectional anterior displacement of the external and internal carotid arteries. Two jugular PGs were found to extend within the lumen of the internal jugular vein. CDS completely depicted carotid body tumors but failed to delineate the high cervical portion of vagal and jugular PGs. MRI allowed us to assess the entire extent of all PGs. Nineteen lesions showed flow voids corresponding to abundant flow signal on CDS: three carotid body tumors appeared hypovascular on CDS and MRI. On CDS, intratumoral flow was directed cranially in carotid and inferiorly in vagal and jugular PGs. CDS and MRI are suitable for classification of cervical PGs as carotid, vagal or jugular PGs based on the topographic relation to the carotid arteries and internal jugular vein. Visualization of the intrinsic tumor vasculature proved an additional distinguishing criterion on CDS. PMID- 11942507 TI - MR imaging, CT, and angiography features of endolymphatic sac tumors: report of two cases. AB - Low-grade adenocarcinoma of endolymphatic sac origin is a rare tumor of the temporal bone. There are some difficulties in its differential diagnosis from other vascular and non-vascular tumors of the temporal bone. However its radiological differentiation from other tumors of the temporal bone is important for surgical planning. We present a report on two endolymphatic sac tumors with some specific radiological findings which can support a correct diagnosis. PMID- 11942508 TI - Conventional MRI and magnetisation transfer imaging of tumour-like multiple sclerosis in a child. AB - Tumefactive multiple sclerosis is a rare entity in children. Differential diagnosis includes other mass lesions such as neoplasm and abscess. A case of tumefactive multiple sclerosis in a child is presented. The open-ring pattern of enhancement on conventional MRI and magnetisation transfer imaging was important for the initial diagnosis and the evaluation of the course of the disease. PMID- 11942509 TI - Which way forward? PMID- 11942510 TI - Reporting news in health--good news, bad news and spin. PMID- 11942511 TI - Awareness of sudden infant death syndrome risk factors among mothers of Pacific infants in New Zealand. AB - AIM: To describe the awareness of Sudden Infant Death Syndrome (SIDS) risk factors among mothers of Pacific infants in New Zealand. METHODS: The data were gathered as part of the Pacific Islands Families Study in which 1376 mothers were interviewed when their infants were six weeks old. Included in this interview were questions designed to examine the mothers' awareness of SIDS risk factors. RESULTS: Over one third (38.8%) of mothers were unable to accurately report a SIDS risk factor, 53.4% reported the risk associated with putting the baby to sleep in a prone position, 31.5% maternal smoking, and 19.5% correctly reported other SIDS risk factors. Lack of awareness of SIDS risk factors was significantly associated with Samoan and Cook Islands Maori ethnicity, being Pacific Islands born, having no post school qualifications, lower household income, not being fluent in English, having more than five children, and not attending antenatal classes. CONCLUSIONS: Despite SIDS prevention efforts, a considerable number of mothers in this cohort reported no awareness of SIDS risk factors. More effective methods are needed to provide consistent SIDS prevention information across Pacific ethnic groups. PMID- 11942512 TI - Associations between ethnicity and obstetric intervention in New Zealand. AB - AIMS: To determine whether the lower rates of obstetric interventions in Maori and Pacific Island women from the New Zealand Ministry of Health obstetric procedures report in 1999 existed also in National Women's Hospital (NWH), Auckland data and if so whether they persisted after controlling for parity and obstetric risk. METHODS: The study population included 43,367 singleton, cephalic deliveries, not preceded by caesarean section at NWH from 1992-1999. Ethnicity was Maori, Pacific Island, or other. Obstetric interventions were explored at two time points: (1) at the initiation of the delivery process: induction of labour, prelabour caesarean section, or spontaneous onset of labour; and (2) at the point of delivery: either caesarean section, operative vaginal delivery, or spontaneous vaginal birth. Independent associations were found by fitting polytomous logistic regression models. RESULTS: 10% of the study population were Maori, 19% Pacific Islanders, and 71% other. Unadjusted analyses showed lower rates of all obstetric interventions for Maori and Pacific Island women. Adjusted analyses showed that rates of induction of labour, prelabour caesarean, and operative vaginal delivery were lower for Maori and Pacific women than for all other ethnicities grouped together. However, caesarean delivery rates overall were not different for Maori or Pacific Island women. CONCLUSIONS: The adjusted analysis did not confirm the association seen in the New Zealand Ministry data between ethnicity and caesarean section. However, induction, prelabour caesarean section, and operative vaginal delivery were less common in Maori and Pacific Island women. PMID- 11942513 TI - Unlocking the numerator-denominator bias. I: Adjustments ratios by ethnicity for 1991-94 mortality data. The New Zealand Census-Mortality Study. AB - AIM: To determine the extent of the under-reporting of Maori and Pacific mortality among 0-74 year olds for the period 1991-94. METHODS: A subset (n=22,578) of highly probable linked 1991 census and 1991-94 mortality records were selected from the 31,635 census-mortality links in the New Zealand Census Mortality Study. The numbers of decedents assigned as Maori, Pacific, and non Maori non-Pacific were compared between mortality and census data. RESULTS: Compared to the death registration form, 29% more 0-74 year old decedents during 1991-94 had self-identified as sole-Maori on the 1991 census (46% for prioritised Maori). This numerator-denominator bias was greater among the young and those living in central and southern New Zealand. Among 0-14, 15-24, 25-44, 45-64, and 65-74 year old decedents, respectively, 91%, 50%, 41%, 26% and 15% more decedents had self-identified as sole-Maori on the 1991 census. For Northern, Midland, Central and Southern regional health authority areas, respectively, 14%, 17%, 81% and 102% more decedents had self-identified as sole-Maori. Among Pacific decedents 68% more 0-74 year old decedents had self-identified as sole-Pacific on the 1991 census (78% for prioritised-Pacific group). This bias for Pacific decedents did not notably vary by age and region. CONCLUSIONS: This study confirms substantial underestimation of Maori and Pacific mortality rates for the period 1991-94, even using the recommended sole-ethnic group denominator. The results from this study should be used to adjust ethnic-specific mortality rates for the early 1990s. Population-based funding formulas that included region specific Maori mortality rates would have particularly disadvantaged central and southern regions. PMID- 11942514 TI - Unlocking the numerator-denominator bias. II: Adjustments to mortality rates by ethnicity and deprivation during 1991-94. The New Zealand Census-Mortality Study. AB - AIMS: Maori and Pacific mortality rates are underestimated due to different recording of ethnicity between mortality and census data--the so-called numerator denominator bias. Ethnicity and deprivation are strongly associated with mortality in New Zealand, but it is unclear what are the independent and overlapping effects of each on health. The objectives of this study were first, to determine the effect of adjusting for numerator-denominator bias on ethnic specific age-standardised all-cause mortality rates among 0-74 year olds during 1991-94: second, to determine the effect of adjusting for numerator-denominator bias on analyses of the independent associations of ethnic group and small area deprivation with all-cause mortality in New Zealand. METHODS: Direct standardisation methods were used to calculate rates of mortality by ethnic and small area deprivation groupings. RESULTS: Unadjusted for numerator-denominator bias, Maori had a 70% and 101% higher standardised mortality rate than non-Maori non-Pacific for males and females, respectively. Adjusting for numerator denominator bias, the excess Maori mortality burden increased to 126% and 158%. For Pacific people, excess mortality increased from -5% and -13% (ie apparently lower mortality rates) to 58% and 54% after adjustment, for males and females respectively. Using data adjusted for numerator-denominator bias, about a third of the Maori to non-Maori non-Pacific disparity in mortality among 0-54 year olds was explained by small area deprivation. Conversely, about a quarter of the mortality gradient by deprivation in New Zealand was explained by ethnic group. CONCLUSIONS: Numerator-denominator bias causes a marked underestimate of the ethnic disparities in mortality in New Zealand for the 1991-4 period, both overall and within strata of deprivation. The distribution of small area deprivation by ethnicity explains some of the ethnic disparities in mortality. PMID- 11942515 TI - Evaluation of the BioSign PSA membrane test for the identification of semen stains in forensic casework. AB - AIM: To evaluate BioSign prostate specific antigen (PSA), a membrane test device used as a clinical aid in the diagnosis of prostate cancer, to determine whether it can be used in forensic laboratories for identifying semen stains. METHODS: Biological fluids were obtained under ethical approval from anonymous consenting donors. BioSign PSA was evaluated in terms of its specificity, sensitivity and cost to replace an ELISA (enzyme linked immunosorbent assay) method of PSA detection. RESULTS: Semen stain extracts and semen diluted 10(5) tested positive with BioSign PSA. Animal semen, other human body fluids and commonly encountered household products tested negative. Anomalous results were observed with semen free condoms containing nonoxynol-9. The cause of these false positive results is not known. CONCLUSIONS: These results and the ease of use of the BioSign PSA kit indicate that it is a valuable addition to forensic laboratories and can adequately replace the ELISA method of PSA detection. BioSign PSA was not suitable for testing condoms for semen. PMID- 11942516 TI - Independent practice associations in New Zealand: a study of governance structures and process. AB - AIMS: To describe the governance structures and processes of four primary care organisations (PCOs) and to evaluate member general practitioners' (GPs) perceptions of the effectiveness of these structures and processes. METHODS: A sample of four PCOs was chosen in 1999, including three independent practitioner associations (IPAs), and a member organisation of CareNet. The chief executive officer of each PCO was interviewed, and the 245 member practitioners were surveyed with a written questionnaire. RESULTS: The response rate to the GP survey was 78.4%. A two tier governance structure was identifiable for all four PCOs: policy board or steering committee, and working committees. In addition each utilised a peer group review process to provide input to some administrative functions. The CareNet PCO emphasised reduced bureaucracy and a loose administrative structure and process. There was a high level of respondent satisfaction (>90%) with their PCO governance processes. CONCLUSIONS: Governance structures and processes have developed within the four PCOs that reflect the needs of member practitioners and according to the requirements of each district and PCO membership. A high level of member satisfaction with their PCO, its governance and processes, was evident. PMID- 11942517 TI - Push enteroscopy: a new service for New Zealand. PMID- 11942519 TI - Patients' rights in the United States: from 'down-under' the situation seems upside-down. PMID- 11942518 TI - Post-market stress syndrome. PMID- 11942520 TI - Mecillinam susceptibility of Auckland urinary isolates. PMID- 11942521 TI - Complementary medicine--again. PMID- 11942522 TI - Data inconsistencies in abstracts in the New Zealand Medical Journal. PMID- 11942523 TI - Molecule capture by olfactory antennules: mantis shrimp. AB - A critical step in the process of olfaction is the movement of odorant molecules from the environment to the surface of a chemosensory structure. Many marine crustaceans capture odorant molecules with arrays of chemosensory sensilla (aesthetascs) on antennules that they flick through the water. We developed a model to calculate molecule flux to the surfaces of aesthetascs in order to study how the size, aesthetasc spacing, and flick kinematics of olfactory antennules affect their performance in capturing molecules from the surrounding water. Since the three-dimensional geometry of an aesthetasc-bearing antennule is complex, dynamically-scaled physical models can often provide an efficient method of determining the fluid velocity field through the array. Here we present a method to optimize the incorporation of such measured velocity vector fields into a numerical simulation of the advection and diffusion of odorants to aesthetasc surfaces. Furthermore, unlike earlier models of odorant interception by antennae, our model incorporates odorant concentration distributions that have been measured in turbulent ambient flows. By applying our model to the example of the olfactory antennules of mantis shrimp, we learned that flicking velocity can have profound effects on odorant flux to the aesthetascs if they operate in the speed range in which the leakiness of the gaps between the aesthetascs to fluid movement is sensitive to velocity. This sensitivity creates an asymmetry in molecule fluxes between outstroke and return stroke, which results in an antennule taking discrete samples in space and time, i.e. "sniffing". As stomatopods grow and their aesthetasc Reynolds number increases, the aesthetasc arrangement on the antennule changes in a way that maintains these asymmetries in leakiness and molecule flux between the outstroke and return stroke, allowing the individual to continue to take discrete samples as it develops. PMID- 11942524 TI - Exciting chaos with noise: unexpected dynamics in epidemic outbreaks. AB - In this paper, we identify a mechanism for chaos in the presence of noise. In a study of the SEIR model, which predicts epidemic outbreaks in childhood diseases, we show how chaotic dynamics can be attained by adding stochastic perturbations at parameters where chaos does not exist apriori. Data recordings of epidemics in childhood diseases are still argued as deterministic chaos. There also exists noise due to uncertainties in the contact parameters between those who are susceptible and those who are infected, as well as random fluctuations in the population. Although chaos has been found in deterministic models, it only occurs in parameter regions that require a very large population base or other large seasonal forcing. Our work identifies the mechanism whereby chaos can be induced by noise for realistic parameter regions of the deterministic model where it does not naturally occur. PMID- 11942525 TI - Dynamics of a food-limited population model incorporating nonlocal delays on a finite domain. AB - In this paper we model and analyse nonlocal spatial effects, induced by time delays, in a diffusion model for a single species confined to a finite domain. The nonlocality, a weighted average in space, arises when account is taken of the fact that individuals have been at different points in space at previous times. We show how to correctly derive the spatial averaging kernels for finite domain problems, generalising the ideas of other investigators who restricted attention to the simpler case of an infinite domain. The resulting model is then analysed and results established on linear stability, boundedness, global convergence of solutions and bifurcations. PMID- 11942526 TI - The universal properties of stem cells as pinpointed by a simple discrete model. AB - The ability of a few stem-cells to repopulate a severely damaged bone marrow (BM) guarantees the stability of our physical existence, and facilitates successful BM transplantations. What are the basic properties of stem cells that enable the maintenance of the system's homeostasis? In the present work we attempt to answer this question by investigating a discrete (in time and phase-space) dynamical system. The model we present is shown to retrieve the essential properties of homeostasis, as reflected in BM functioning, namely, (a) to produce a constant amount of mature cells, and (b) to be capable of returning to this production after very large perturbations. The mechanism guaranteeing the fulfillment of these properties is extrinsic--negative feedback control in the micro-environment -and does not need additional stochastic assumptions. Nevertheless, the existence of a simple intrinsic control mechanism, a clock which determines the switch to differentiation, ascertains that the system does not admit non-trivial extinction states. This result may help clarifying some of the controversy about extrinsic versus intrinsic control over stem cell fate. It should be stressed that all conclusions are valid for any system containing progenitor and maturing cells. PMID- 11942527 TI - Wave propagation in discrete media. AB - We have considered infinite systems of nonlinear ODEs on the one-dimensional integer lattice which describes the activity in an excitatorily coupled network of excitable cells. For an ideal nonlinearity, we calculated the speed of propagation of an activity and derived the condition for its existence. We also studied the existence and stability of the traveling wave solution and gave, in the simplest case, its explicit expression. We established that some unstable traveling waves lead to propagation with an enlarging profile defined by a front velocity and a wake velocity. We generalized some results to inhomogeneous medium and network with long range connections. PMID- 11942528 TI - Mode-doubling and tripling in reaction-diffusion patterns on growing domains: a piecewise linear model. AB - Reaction-diffusion equations are ubiquitous as models of biological pattern formation. In a recent paper we have shown that incorporation of domain growth in a reaction-diffusion model generates a sequence of quasi-steady patterns and can provide a mechanism for increased reliability of pattern selection. In this paper we analyse the model to examine the transitions between patterns in the sequence. Introducing a piecewise linear approximation we find closed form approximate solutions for steady-state patterns by exploiting a small parameter, the ratio of diffusivities, in a singular perturbation expansion. We consider the existence of these steady-state solutions as a parameter related to the domain length is varied and predict the point at which the solution ceases to exist, which we identify with the onset of transition between patterns for the sequence generated on the growing domain. Applying these results to the model in one spatial dimension we are able to predict the mechanism and timing of transitions between quasi-steady patterns in the sequence. We also highlight a novel sequence behaviour, mode-tripling, which is a consequence of a symmetry in the reaction term of the reaction-diffusion system. PMID- 11942529 TI - Dynamics of herbivore-plant-pollinator models. AB - A mathematical model for the plant-pollinator interaction for which the relationship is not obligate is discussed. The dynamics of the system depend on both the energetic rewards and the specificity of the pollinator to the plant. Based on this model, the interaction between herbivore, plant and pollinator is proposed and the possible effect of reducing pollinator visitation rate due to the presence of herbivore is considered. It is shown that this reduction mechanism has no effect on the qualitative behavior of the model. By using these results, the possible mechanism of enhancing pollinator visitation rate accompanied by the herbivore upon the system is also examined. It is demonstrated that such a mechanism in general can promote the persistence of this three population interaction. PMID- 11942530 TI - Integrodifference equations, Allee effects, and invasions. AB - Models that describe the spread of invading organisms often assume no Allee effect. In contrast, abundant observational data provide evidence for Allee effects. We study an invasion model based on an integrodifference equation with an Allee effect. We derive a general result for the sign of the speed of invasion. We then examine a special, linear-constant, Allee function and introduce a numerical scheme that allows us to estimate the speed of traveling wave solutions. PMID- 11942531 TI - A status-based approach to multiple strain dynamics. AB - We present and investigate a new model for cross-immunity. Past models classify hosts according to their infection history. Here we represent hosts through their status: their current ability to respond to strains. This framework allows a different, a wider, and a more biologically interpretable range of forms of cross immunity to be studied. Using this new form of cross-immunity we then consider a previously studied case of four strains, each of which confers partial immunity to two of the others. In this interesting special case, with applications to the genetic maintenance of strain diversity, we can make substantial analytical progress. We present methods for exploiting the symmetries of the system to show that only a particular invariant subspace need be considered for characterizing the dynamics of the whole system. A complete bifurcation structure is given for this subspace. In contrast to systems previously studied, this system does not exhibit sustained oscillations for any set of parameter values. PMID- 11942532 TI - Density-dependent birth rate, birth pulses and their population dynamic consequences. AB - In most models of population dynamics, increases in population due to birth are assumed to be time-independent, but many species reproduce only during a single period of the year. We propose a single-species model with stage structure for the dynamics in a wild animal population for which births occur in a single pulse once per time period. Using the discrete dynamical system determined by the stroboscopic map, we obtain an exact periodic solution of systems which are with Ricker functions or Beverton-Holt functions, and obtain the threshold conditions for their stability. Above this threshold, there is a characteristic sequence of bifurcations, leading to chaotic dynamics, which implies that the dynamical behaviors of the single species model with birth pulses are very complex, including small-amplitude annual oscillations, large-amplitude multi-annual cycles, and chaos. This suggests that birth pulse, in effect, provides a natural period or cyclicity that allows for a period-doubling route to chaos. PMID- 11942533 TI - The role of dietary and plasma lipids in childhood atherogenesis. AB - Maria Daria Haust, MD, has made many significant contributions to our understanding of atherogenesis in children, particularly in associating the earliest lesions (fatty spots and streaks) with the normal growth and remodeling of arteries. It is my privilege to link her early work to more recent findings which show that the early lesions seen in the arteries of children before puberty bear no relationship to the risk of atherosclerosis in later life. Furthermore, present evidence supports the view that intervening in childhood (2-15 years) with low-fat low-cholesterol diets or even worse, lipid-lowering drugs to prevent atheroslerotic plaques in adulthood is wasted effort. Overzealous parents may unwittingly induce malnutrition in their children and many children with restricted access to palatable foods, will yearn for them even more as they become older leading to over weightness. Pediatricians from various scientific bodies around the world vary in their advice to mothers regarding diets for children. The program adopted by Health Canada on the advice of pediatricians in that country, which is also supported by the European Society of Pediatrics, Gastroenterology and Nutrition, recommends that the fat content of diets for children should be tapered gradually from 40% of energy at 2 years of age to 30% of energy at the conclusion of linear growth (late adolescence). PMID- 11942534 TI - Cholesterol and development: the RSH ("Smith-Lemli-Opitz") syndrome and related conditions. AB - The half-century of lipophobia in the United States may be abating with some return of sanity on the discussion of health and dietary fat [Taubes, 2001]. The youngest victims of this collective, decades long madness are those infants deprived for one reason or another of breast milk. They are unable to speak for themselves at a time of greatest need for cholesterol during growth, the most critical period of myelination of central and peripheral nervous system, formation of bone and bile, and of every steroid hormone. Some of the commercial formulas they are fed contain only 1 or 2 mg of cholesterol per 100 g edible portion contrasted with almost 14 mg in breast milk. One can only hope that the confidence in their endogenous ability to synthesize sufficient amounts of cholesterol is not misplaced. Pediatric pathology has learned that when this endogenous ability fails during embryogenesis on the basis of mutations in the postsqualene biosynthesis of cholesterol, a startling variety of developmental pathology may present itself ranging from lethal forms of "idiopathic" hydrops, microcephaly with cerebral dysgenesis and dysmyelinization, agenesis of corpus callosum, cerebellar vermis dysgenesis, cataracts, cleft palate, many different forms of congenital heart defect, pyloric stenosis and/or Hirschsprung dysganglionosis, adrenal (cortical) insufficiency, cholestatic liver disease, limb malformations, and genital ambiguity in genetic males. Population genetic considerations suggest a hypothetical birth prevalence of the RSH (so-called Smith-Lemli-Opitz) syndrome, the commonest of these Garrodian errors of cholesterol biosynthesis, of 1/2500; since only about 1/15,000 to 1/20,000 homozygotes are liveborn and biochemically confirmed, over 80% prenatal or perinatal mortality must occur and deserves the most discerning of services from birth attendants, perinatologists, neonatologists, and fetal/pediatric pathologists. An easy, reliable, economical biochemical test for the presence of 7-dehydrocholesterol is available and the commonest mutation, the IVS8-1G-->C mutation, is quickly and reliably tested for molecularly. Thus, the successful diagnosis, even after death, will contribute substantially to correct genetic counseling, carrier detection, prenatal diagnosis, and treatment in those known to be affected prenatally andplanned to be liveborn. Thus, developmental pathology plays an integral, vital role in preventive medicine. PMID- 11942535 TI - Pathology of lethal fetal growth retardation syndrome with aminoaciduria, iron overload, and lactic acidosis (GRACILE). AB - Autopsy study of 17 newborn infants with lethal autosomal recessive disease presenting as growth retardation with lactic acidosis, Fanconi aminoaciduria, and hepatic hemosiderosis is reported. The patients succumbed between day 1 and 4 months of life; 9 patients died within the first month. All patients showed severe pathologic changes of liver with cholestasis in all livers. Extensive accumulation of stainable iron of the hepatocytes was present in 9/17 autopsy tissues and in two biopsy specimens. Moderate to abundant iron storage in the Kupffer cells was seen in all liver specimens. The amount of hepatocytic iron was high in livers up to 1 month of age and decreased thereafter. The general features and liver findings of this disorder suggest the name Growth Retardation Aminoaciduria Cholestasis Iron Overload, Lactacidosis and Early Death (GRACILE, OMIM 603358). Calcified concrements were seen in the medulla of 13/16 kidney specimens. Pancreas of 13/14 patients showed interstitial fibrosis and exocrine atrophy. Various pathologic findings such as renal tubular dysgenesis, paucity of hepatic bile ducts and iron storage in the macrophages of spleen and pulmonary alveoli were observed in some cases. Previous extensive clinical genetic and laboratory investigations have revealed that the patients had a previously unrecognized genetic disease. It is inherited as an autosomal recessive trait. The gene locus is 2q33-37. The basic defect of the disease remains unknown. PMID- 11942536 TI - Cardiovascular causes for sudden infant death. AB - Although sudden infant death syndrome (SIDS) is a cause for sudden infant death, other causes should be ruled out before diagnosing SIDS. Cardiac causes for sudden infant death include viral myocarditis, congential heart disease particularly congential aortic stenosis, endocardial fibroelastosis, and anomalous origin of the left coronary artery from the pulmonary artery. Other cardiac conditions that may result in sudden death include rhabdomyomas of the heart in tuberous sclerosis and conduction system disorders. The most frequent conduction system disorders resulting in sudden death include histiocytoid cardiomyopathy, congential heart block that may be associated with maternal lupus erythematosus, arrhythmogenic right ventricular dysplasia, noncompaction of the left ventricle, and long QT syndromes. PMID- 11942537 TI - Natural history and risk factors of atherosclerosis in children and youth: the PDAY study. AB - The Pathobiological Determinants of Atherosclerosis in Youth (PDAY) study was organized to document the natural history of athersclerosis and to determine the relation of cardiovascular risk factors to atherosclerosis in young subjects. Pathology laboratories in 15 centers collected coronary arteries, aortas, and other tissues from over 3,000 subjects age 15 to 34 who died of external causes between 1987 and 1994. The extent, prevalence, and topography of arterial lesions were evaluated and risk factors were analyzed in a central laboratory. Postmortem risk factors included serum lipoproteins, serum thiocyanate (smoking), glycohemoglobin (diabetes), thickness of panniculus adiposus and body mass index (obesity), changes in small renal arteries (hypertension), and apoprotein isoforms. The PDAY study confirmed the origin of atherosclerosis in childhood, showed that progression toward clinically significant lesions may occur in young adulthood and demonstrated that the progression of atherosclerosis is strongly influenced by coronary heart disease (CHD) risk factors. Recent PDAY studies have shown that a significant number of advanced coronary artery lesions have microscopic qualities associated with susceptibility to rupture and that CHD risk factors are associated with the development of these characteristic microscopic qualities. The PDAY archive continues to provide an important resource for new investigators throughout the world that contribute to the understanding of atherosclerosis, the underlying cause of most cardiovascular disease and the leading cause of debilitating illness and death in this country. The PDAY findings emphasize the need to modify risk factors in young people to retard the development of atherosclerotic lesions, particularly clinically significant lesions. Thus, true primary prevention of atherosclerosis must being in childhood or early adolescence. PMID- 11942538 TI - Chronic disease from early malnutrition. PMID- 11942539 TI - Cardiovascular involvement in metabolic diseases. AB - Cardiomyopathies are a group of disorders that result in dysfunction of the contractility of the myocardium anda decrease in cardiac function. Cardiomyopathies may be primary or idiopathic or secondary due to an underlying definable systemic disorder. Cardiomyopathies may be further defined as according to the World Health Organization (WHO) as: 1) congestive or dilated; 2) obstructive as in hypertrophic cardiomyopathy; 3) restrictive as in infiltrative disorders such as storage diseases. Two additional categories have been added that include arrhythmogenic right ventricular cardiomyopathy and unclassified cardiomyopathies. A number of metabolic disorders result in cardiomyopathy and are frequently the cause of death in these disorders. PMID- 11942540 TI - Comparison of trabeculectomy with mitomycin C with or without phacoemulsification and lens implantation. AB - PURPOSE: To compare the results of combined trabeculectomy with phacoemulsification and posterior chamber intraocular lens (IOL) implantation to those of trabeculectomy alone using mitomycin C (MMC) application intraoperatively in all cases. PATIENTS AND METHODS: A retrospective comparative study of consecutive patients was conducted on two groups: 102 eyes of 90 patients studied in the combined procedure group, and 33 eyes of 30 patients in the trabeculectomy alone group. RESULTS: Both groups showed a significant decrease in IOP. The combined group had a change from 21.5+/-5.8 mm HG preoperative to 14.73+/-3.44 mm HG postoperative, P=0.0001; the trabeculectomy group changed from 24.2+/-7.5 mm HG preoperative to 12.46+/-3.86 mm HG postoperative, P=0.0001. This represents a 31.5% reduction in IOP in the combined group versus a 48.5% reduction in the trabeculectomy alone group (P=0.0001). The follow-up time was longer in the trabeculectomy group (trabeculectomy group, 22.6+/-13.3 months; combined group, 14.2+/-8.0 months), P=0.0014. There were 97 eyes in the combined group (95%) and 32 eyes (97%) in the trabeculectomy group that had an IOP of less than 20 mm HG at the end of follow up. Postoperatively, the two groups showed similar significant reductions in the number of antiglaucomatous medications used (combined group, 0.82+/-1.0 compared with 2.65+/-0.84 preoperatively, P=0.0001; trabeculectomy group, 0.76+/-1.2 compared with 2.7+/-0.95 preoperatively, P=0.0001). There were no cases of bleb leakage in the combined group and two cases (6%) in the trabeculectomy group. CONCLUSION: The reduction of IOP is significantly larger after trabeculectomy alone than after the combined procedure; however, the functional and anatomical results of the combined procedure of phacoemulsification, posterior chamber IOL implantation, and trabeculectomy with MMC application were as good as those of trabeculectomy alone with MMC. PMID- 11942541 TI - 5-Fluorouracil-needling and paracentesis through the failing filtering bleb. AB - BACKGROUND AND OBJECTIVE: Standard needle revision of the filtering bleb after glaucoma filtration surgery has many disadvantages. This study reports the technique and outcome of a modified needling of the failing bleb. PATIENTS AND METHODS: Consecutive patients who presented with increased intraocular pressure (lOP) and open internal ostium were included in this prospective study. When their IOP rose to levels >20 mm Hg despite various therapeutic regimens and visibility of the sclerostomy site was unclear, the modified needling technique was conducted. In each needling, a total dose of 1 mg (0.1 mL) of 5-fluorouracil (FU) was administered subconjunctivally, adjacent and into the bleb. The needle was then passed via the bleb, anterior to the sclera, through the anterior limbus to penetrate intracamerally. Standard treatment to inhibit bleb scarring followed. RESULTS: Twelve 5-FU-needle revisions were performed on 8 eyes. The first needle revision was performed 51.1+/-36.2 days (range, 5-117) postoperatively. IOP decreased from 31.0+/-5.7 (range, 25-43) mm Hg with 0.6+/ 0.9 (range, 0-2) hypotensive medications to 15.5+/-2.6 mm Hg (range, 12-19) with 0.9+/-1.1 medications (range, 0-3), 11.3+/-4.9 months (range, 8-23) after the only or second (last) needling (P=0.012). In all eyes, the filtering bleb became diffused; in 7 eyes, it remained diffusely elevated or shallow-diffused. CONCLUSION: Relatively few needle revisions of the filtering bleb by a modified approach that bypassed the activated scarring tissue contributed to the restoration of IOP control. It is possible that the 5-FU-needle revision may offer advantages over the standard approach. A larger sample and a controlled study are required to validate these considerations. PMID- 11942542 TI - Reproducibility of retinal nerve fiber layer thickness measurements using scanning laser polarimetry in pseudophakic eyes. AB - OBJECTIVE: To assess the reproducibility of retinal nerve fiber layer measurements in pseudophakic normal and glaucomatous eyes using scanning laser polarimetry (GDx, Laser Diagnostic Technologies, Inc., San Diego, CA). PATIENTS AND METHODS: Normal and glaucomatous patients with polymethylmethacrylate posterior chamber intraocular lenses that satisfied entry criteria underwent imaging by two experienced operators. Eyes with posterior capsule opacification or vision less than 20/30 were excluded. Baseline images (mean pixel SD less than 8 mm) were obtained on 3 separate days within a 7-week period. Reproducibility, defined as the pooled within eye variance of these 3 measurements and the coefficient of variation for 12 retardation parameters generated by GDx software were calculated. RESULTS: Eighteen eyes (11 glaucoma, 7 normal) of 15 patients (7 female, 8 male) were enrolled (mean age 78 +/- 6 years). Among glaucomatous eyes, the average mean deviation and corrected pattern standard deviation using achromatic automated perimetry (Zeiss-Humphrey, Dublin, CA) was -3.8 +/- 1.5 dB (range, -1.89 to -5.04 dB) and 4.9 +/- 3.3 dB (range 0 to 11.05 dB), respectively. Coefficient of variation was 10% or less for all retardation parameters except ellipse modulation (20.2%) and neural network number (12.4%). Glaucomatous and normal eyes had similar variability for 8 of 12 (66.7%) retardation parameters. Inferior ratio, ellipse modulation, and superior ratio were significantly less variable in glaucomatous eyes (P = 0.007, 0.02, and 0.04 respectively) than normal eyes. Superior integral was more variable in glaucomatous eyes (P = 0.03). CONCLUSION: Retardation measurements may be obtained in pseudophakic eyes with acceptable reproducibility. Normal eyes and eyes with mild glaucomatous damage have similar variability for most retardation parameters. PMID- 11942543 TI - Topographic mapping of glaucomatous visual field defects to scanning laser polarimetry of the peripapillary nerve fiber layer. AB - BACKGROUND AND OBJECTIVE: To map retinal sensitivity values within glaucomatous visual field defects (VFD) to corresponding values of scanning laser polarimetry (SLP) of the peripapillary retinal nerve fiber layer using the Wirtschafter disc sector classification. METHODS: Eyes with glaucomatous VFD (N = 28) underwent SLP and Humphrey automated perimetry. The Wirtschafter disc sector classification was used to obtain SLP values corresponding to VFD. The average and modulated SLP values were calculated and correlated with VFD. RESULTS: No significant correlation was found between retinal sensitivity and the Wirtschafter disc sector SLP values, using average values (P =.904) or modulated values (P =.409). CONCLUSION: Despite use of modulation parameters customized to correspond with the Wirtschafter disc sector classification, topographic mapping of retinal sensitivity levels within glaucomatous VFD to corresponding SLP values was not possible in this group of glaucomatous eyes. PMID- 11942544 TI - Relation between retinal thickening and clinically visible fundus pathologies in mild nonproliferative diabetic retinopathy. AB - BACKGROUND AND OBJECTIVE: To determine the association of retinal thickening (RT) with clinically observable retinal pathologies in eyes with mild nonproliferative diabetic retinopathy. PATIENTS AND METHODS: Using an objective quantitative imaging method (Retinal Thickness Analyzer), the ratio relative to normal RT (RTI) was measured in 23 eyes with and 35 eyes without clinically observable diabetic fundus pathology. RTI was analyzed in relation to presence of mild diabetic retinal lesions in the +/-0.5 mm vicinity. RESULTS: The percent of eyes with RTI significantly above normal values did not differ significantly between eyes with and without retinopathy (30% vs 34%). Mean RTI was not associated with local presence of microaneurysms (P=0.92), soft exudates (P=0.55), or retinal hemorrhages (P=0.31). Areas without hard exudates had significantly greater mean RTI (1.10) than areas with exudates (0.97, P=0.009). CONCLUSION: In diabetic patients with mild retinopathy, areas with and without clinically observable retinal pathologies had similar retinal thickness. We conclude that clinical strategies for detection of retinal thickening should not be limited to areas with visible fundus pathologies. PMID- 11942545 TI - Treating cytomegalovirus retinitis-related retinal detachment by combining silicone oil tamponade and ganciclovir implant. AB - BACKGROUND AND OBJECTIVE: With the efficacy of pars plana vitrectomy and silicone oil infusion in treating cytomegalovirus (CMV) retinitis-related retinal detachment and the success of the ganciclovir implant in controlling CMV retinitis, we sought to evaluate the possible benefits of combining these two procedures in one surgical operation. PATIENTS AND METHODS: A retrospective review of 10 patients was conducted. Each patient was diagnosed with a CMV retinitis-related retinal detachment and treated with pars plana vitrectomy and silicone oil infusion, with simultaneous placement of a ganciclovir implant. Parameters evaluated included location of retinal detachment, reattachment rate, pre- and post-operative Snellen visual acuity, pre- and post-operative CMV retinitis activity and location, and complications of the combined procedure. RESULTS: Overall anatomic reattachment was achieved in all 10 patients. Four patients presented with macular involvement of their retinal detachments. Three of these patients experienced significant post-operative improvement in visual acuity. Surgery preserved visual acuity in the 6 patients who presented with macula attached. Best postoperative acuity was better than or equal to 20/100 in 7 (70%) patients. All 3 CMV retinitis patients with inactive retinitis preoperatively remained free of retinitis for the duration of follow up. At last follow up, 8/10 (80%) showed no active CMV retinitis and no patients experienced progression of their retinitis. CONCLUSIONS: Results of this series indicate that patients benefit from excellent anatomic reattachment rates, preservation or improvement of visual acuity in most cases, and extended control of their CMV retinitis. Combining the two procedures appears viable. Further study is warranted to assess definitive anatomic and functional outcomes resulting from this new technique. PMID- 11942546 TI - Ocular toxicity of intravitreal tacrolimus. AB - BACKGROUND AND OBJECTIVE: To investigate the ocular toxicity of intravitreally administered tacrolimus, a drug with potent immunosuppressive activity. METHODS: To evaluate toxicity, tacrolimus was injected into the midvitreous cavity of 20 eyes of New Zealand pigmented rabbits at concentrations of 10, 50, 100, 250, 500, and 1000 microg. Control eyes received balanced salt solution. Eyes receiving 1000 microg were given injections of 0.2 mL solution; all others, including controls, received 0.1 mL. Rabbits were examined before the injections by slit lamp biomicroscopy, indirect ophthalmoscopy, and an electroretinography test (ERG) was performed. The animals were followed up to 14 days postinjection by clinical examination and ERG. The animals were killed and the eyes were enucleated and processed for light microscopy. RESULTS: No evidence of a retinal toxic reaction was seen in the eyes receiving 10 or 50 microg of tacrolimus. One out of 4 eyes that received 100 microg of the drug developed a vitreous reaction. All eyes treated with 250 microg or more developed vitreous reaction. One eye injected with 1000 microg of the drug developed occlusion of the temporal retinal vessels. Electroretinography showed decreasing b-wave amplitude with both dark- and light-adapted stimulus in the 500 and 1000 microg groups, and it was normal in the other groups. Histopathologic sections showed mild disorganization of the retina only at the 500 and 1000 microg dosage. CONCLUSIONS: Doses of 10 and 50 microg of tacrolimus are nontoxic to rabbit eyes. Only transient vitreous opacities were observed in the groups that received 100 and 250 microg. Intravitreal doses of 500 and 1000 microg of tacrolimus proved to be toxic to the retina. PMID- 11942547 TI - Paramacular Von Hippel angioma with tractional macular detachment. AB - A rare case of paramacular angioma with tractional macular detachment that was managed successfully with laser, followed by surgical intervention with good visual outcome, is presented. A 23-year-old female patient with paramacular Von Hippel angioma was treated with argon laser photocoagulation. Three-months following photocoagulation, vitrectomy with epiretinal tractional membrane removal was attempted successfully. Visual acuity improved from a preoperative level of 20/200 to 20/50 at the 6 months postoperative period. No recurrence of epiretinal membrane or exudation was observed during the follow-up period. Photocoagulation, followed by surgical intervention for removal of epiretinal traction membranes, resulted in relieving the tractional macular detachment and recovering useful visual acuity in paramacular Von Hippel angioma. PMID- 11942548 TI - Bilateral massive choroidal hemorrhage secondary to Glanzmann's syndrome. AB - Characteristics of intraocular bleeding and its management in association with blood dyscrasias are discussed. We present a patient with massive bilateral choroidal hemorrhage secondary to Glanzmann's syndrome. Magnetic resonance imaging and ultrasonographic findings were ordered. During the clinical course, bilateral intravitreal hemorrhage and tractional retinal detachment occurred. Left pars plana vitrectomy was performed under general anaesthesia. The procedure was unsuccessful because of intraoperative uncontrolled bleeding. PMID- 11942549 TI - Bilateral macular hole formation in a patient with retinitis pigmentosa. AB - A 47-year-old woman known to have retinitis pigmentosa (RP) developed macular holes in each eye. Examination and fluorescein angiography demonstrated a partial thickness macular hole in the right eye and a full thickness one in the left eye. It was concluded that macular hole formation can occur in patients with RP, but the exact cause of macular hole development in these patients is uncertain. PMID- 11942550 TI - Endophotocoagulation to retinal pigment epithelium as an adjuvant therapy in the management of retinal detachment caused by a highly myopic macular hole. AB - Retinal detachment secondary to a highly myopic macular hole remains a particular surgical challenge. Pars plana vitrectomy, endophotocoagulation at the base of a macular hole in conjunction with fluid-gas exchange was performed in 4 consecutive patients with a mean refraction of -13.8D (range, -8D-(-)18D). Mean axial length was 27.5 mm (range, 26.0-29.0 mm). Mild atrophy of the retinal pigment epithelium was present in 3 patients while 1 patient had moderate atrophy. The mean follow-up period was 12 months (range, 6-18 months). The overall primary anatomical success rate was 75%. All 3 eyes with mild retinal pigment epithelium atrophy had retinal reattachment after one operation. PMID- 11942551 TI - Melanocytoma of the choroid: angiographic and histopathologic findings. AB - A juxtapapillary choroidal melanocytoma that was clinically confused with a choroidal melanoma is described. The clinical examination of a 40-year-old female patient included ultrasonography, fluorescein, and indocyanine green angiography. Histopathologic and immunohistochemical study of the enucleated eye resulted in the identification of a markedly pigmented, 3.8 mm elevated choroidal melanocytic tumor observed nasally to the disk. A-scan ultrasonography showed a high initial spike with a low to medium internal reflectivity with decreasing amplitude. On fluorescein angiography, the tumor was surrounded by a rim of retinal pigment epithelium depigmentation with a well-defined comet-tail. The lesion was hypofluorescent in the early phases of the indocyanine green angiogram, but from the late venous phase appeared ill-defined hyperfluorescence. Histopathologic study revealed a juxtapapillary choroidal melanocytoma The pigmented tumor cells stained positive for HMB45 and vimentin but negative for S100, GFAP, NSE, and MNF116. No tumor cells stained positive for the proliferation marker Ki67. These findings conclude that choroidal melanocytoma may mimick choroidal melanoma. Abnormalities on fluorescein angiography may be a clinical hint to suspect a pigmented tumor different from a choroidal melanoma. PMID- 11942552 TI - Acute lymphocytic leukemia presenting as masquerade syndrome. AB - An 11-year-old child presented with plastic iridocyclitis and hypopyon. Subsequent paracentesis revealed leukemic infiltrates. A bone marrow biopsy confirmed the diagnosis of acute lymphocytic leukemia. Because of this unusual case of leukemic hypopyon, we advocate anterior chamber paracentesis and pediatrician referral in all cases of refractory uveitis. PMID- 11942553 TI - Electrical cataract: a case study. AB - A case of bilateral cataract sustained after electrical trauma was seen in a referral practice. We examined a young patient with dense anterior and posterior subcapsular cataract in his right eye and pseudophakia in his left eye who gave a history of gradually progressive painless loss of vision after sustaining injury from a high voltage electric current. Extracapsular cataract extraction (ECCE) with posterior chamber intraocular lens (PCIOL) implantation was performed in his right eye. Visual rehabilitation in both eyes after ECCE with PCIOL implantation was good with the patient attaining uncorrected visual acuity of 20/30 in both eyes postoperatively. It appears that the surgical results and visual rehabilitation after ECCE with PCIOL implantation in patients with electrical cataract without any other ocular damage is excellent. PMID- 11942554 TI - Modified grey line split with anterior lamellar repositioning for treatment of cicatricial lid entropion. AB - To determine the success rate of surgery of modified grey line split with anterior lamellar repositioning in patients with cicatricial lid entropion and to determine the risk factors of failure of the procedure, 40 patients (84 lids) with either lid involvement caused by cicatricial lid entropion of different etiologies were enrolled in this study. All the lids were operated on using the technique of modified grey line split and anterior lamellar repositioning. The success of the procedure was assessed by restoration of anatomical and physiological functioning of the lid without any residual symptom to the patient. Patients were examined initially at weekly intervals for 1 month and subsequently followed up at 2, 3, and 12 months following surgery. Among the various causes for cicatricial lid entropion, infectious etiology (72/84 lids) was found to be the most common one. A success rate of modified grey line split with anterior lamellar repositioning was 88.09% (74/84 lids). The underlying etiology of cicatricial lid entropion was the sole predictor of failure of surgery. Those with the etiology of infection had more than 6 times the odds of surgery failure (OR: 6.73; 95% CI: 2.79-16.73) as compared to a patient without infectious etiology. The role of other factors such as the age of the patient, degree of entropion, previous entropion surgery, the lid (upper or lower) involved, irregular lid margin, and defective lid closure were statistically insignificant. The underlying etiology of cicatricial lid entropion is the only risk factor that significantly influences the outcome of surgery with this technique. Otherwise, this procedure gives good results with fewer complications in patients with cicatricial lid entropion. PMID- 11942555 TI - Decompression retinopathy following filtration surgery. PMID- 11942556 TI - Comparison of double-plate Molteno and Ahmed glaucoma valve in patients with advanced uncontrolled glaucoma. AB - BACKGROUND AND OBJECTIVE: To compare double-plate Molteno (DPM) with the Ahmed glaucoma valve (AGV) in the treatment of eyes with complicated glaucoma. PATIENTS AND METHODS: This matched, retrospective, case-control study was based on diagnosis, number of previous operations, and age. There were 30 patients in each group and only patients with a minimum of 6 months follow up were included. Double-plate Molteno or Ahmed glaucoma valve insertion was performed on each patient to control intractable glaucoma. Success was defined as IOP <22 mm Hg and >4 mm Hg on the last two visits, a decrease of no more than 2 lines in the visual acuity, and no additional surgical intervention to control IOP. RESULTS: The Kaplan-Meier estimated probability of success at 12 and 24 months was 73% and 56% with DPM and 60% and 50% for AGV (P = 0.72). Mean IOP measured 13.36 +/- 5.2 mm Hg following DPM and 16.7 +/- 5.6 mm Hg following AGV at 12 months (P = 0.026) and 13.3 +/- 5.1 mm Hg with DPM and 19 +/- 5.8 mm Hg with AGV at 24 months (P = 0.009). Of the patients involved in the study, 83.5% exhibited hypertensive phase (HP) of AGV vs 43.5% of DPM (P = 0.04). AGV had a tendency to fail earlier (5 months +/- 7 following AGV vs 13 months +/- 13 following DPM, P = 0.07, t-test). Patients with either a second valve insertion or valve removal were 7/30 of AGV vs 1/30 in DPM (P = 0.05). Stent removal was needed by 10/30 DPM and 8/30 AGV had needling/5-FU. CONCLUSIONS: The percentage of patients free from failure was similar between the two groups at 36 months. DPM is associated with lower mean IOP in the long term compared to AGV. PMID- 11942557 TI - Macrophage arginine metabolism to ornithine/urea or nitric oxide/citrulline: a life or death issue. AB - Macrophages can metabolize arginine to nitric oxide in quantities that inhibit pathogens or nearby host cells. They can instead metabolize arginine to ornithine (a precursor of polyamines and collagen) in quantities that stimulate pathogens or nearby host cells. Macrophages are essentially the only circulating cells that can make these life or death decisions with arginine. Macrophages expressing these destructive or constructive phenotypes have been termed M-1 or M-2 because they also stimulate TH1 or TH2 responses, respectively. Factors that influence whether a macrophage expresses the M-1 or M-2 phenotype and the real or potential impact on immune responses and other host processes are discussed. PMID- 11942558 TI - Interleukin-10 in the brain. AB - Interleukin (IL)-10 is synthesized in the central nervous system (CNS) and acts to limit clinical symptoms of stroke, multiple sclerosis, Alzheimer's disease, meningitis, and the behavioral changes that occur during bacterial infections. Expression of IL-10 is elevated during the course of most major diseases in the CNS and promotes survival of neurons and all glial cells in the brain by blocking the effects of proapoptotic cytokines and by promoting expression of cell survival signals. Stimulation of IL-10 receptors regulates numerous life- or death-signaling pathways--including Jak1/Stat3, PI 3-kinase, MAPK, SOCS, and NF kappaB--ultimately promoting cell survival by inhibiting both ligand- and mitochondrial-induced apoptotic pathways. IL-10 also limits inflammation in the brain; it does so by three major pathways: (1) reducing synthesis of proinflammatory cytokines, (2) suppressing cytokine receptor expression, and (3) inhibiting receptor activation. Finally, IL-10 induces anergy in brain infiltrating T cells by inhibiting cell signaling through the costimulatory CD28 CD80/86 pathway. The multiple functions of IL-10 in the brain will create new and intriguing vistas that will promote a better understanding of neurodegenerative diseases. These discoveries could lead to development of innovative approaches for the use of antiinflammatory cytokines in major debilitating diseases of the CNS. PMID- 11942559 TI - Induction and regulation of T-cell priming for contact hypersensitivity. AB - Contact hypersensitivity (CHS) is a T-cell-mediated immune response to cutaneous sensitization and subsequent challenge with haptens such as dinitrofluorobenzene and oxazolone. Clinically, contact sensitivity, also called allergic contact dermatitis, is a frequently observed dermatosis in industrialized countries. Experimental CHS in mice has been used by many laboratories as a model of T-cell mediated immune responses to antigens deposited onto the skin to study the priming, development, and function of effector and regulatory T-cell components during these responses. In this article we discuss the mechanism of T-cell priming by hapten-presenting Langerhans cells and how the priming environment influences the development of these hapten-specific T cells to different functional phenotypes during sensitization for the CHS response. Finally, we propose a model of negative regulation of the CHS response by T-cell components that are coincidentally primed with the effector T cells mediating the response. Overall, these aspects indicate a unique immune response mediated and regulated by specialized antigen-presenting cells and T-cell populations. PMID- 11942560 TI - Readiness for Medicaid managed care reform initiatives: a profile of rural family physicians. AB - Medicaid managed care is now an important factor in the financing of rural health care delivery. The participation of rural family physicians in Medicaid managed care is vital for the rural poor to access health services. This study examined 855 family physicians practicing in nonmetropolitan counties across the United States to determine their readiness to participate in Medicaid managed care. Physicians were asked about their experience with prepaid programs and the factors that would influence their participation in such a program. A shortage of health care providers and low reimbursement rates were most frequently cited as barriers to successful implementation. Physicians who had participated in prepaid programs in the past but were no longer participating had the most negative opinions about the potential for Medicaid managed care programs to enhance care for the poor in their communities. Overall, physicians reported potential for the program to improve access and quality of care, but they also expressed reservations about the financial and administrative effects on their practices. These results reveal that negative attitudes were associated with prepaid programs that failed to meet expectations, but physicians also expressed an optimism about the potential to serve the poor within a managed care model. PMID- 11942561 TI - Analysis of tractor-related deaths in North Carolina from 1979 to 1988. AB - This descriptive study examined 342 fatalities involving tractors and attachments that occurred during a 10-year period in North Carolina from 1979 to 1988. Reports of the North Carolina Office of the Chief Medical Examiner and medical examiner's certificates of death were reviewed using the public health model of causation. Data were collected on characteristics of thefatally injured (host), including age, occupation, blood alcohol concentration and length of survival; characteristics of the injury event (environment), including type of accident, weather conditions and terrain; and characteristics of the agent, including both the tractor and attachments, if any, in place at the time of injury. Ninety-eight percent of the victims were male. Victims ages 65 and older accounted for 38 percent of fatal injuries, and 11 percent were 18 years of age or younger. Only slightly more than one-half of the victims (54 percent) were full- or part-time farmers. Forty-four percent of the deaths were to nonfarmers who were not in the act of farming. Nineteen percent of tested victims had a detectable blood alcohol level. Almost three-fourths of the victims died in the first hour after injury and 87 percent within the first 24 hours. In the majority of accidents, the tractor operator was a victim of a rollover or runover. This study provided detailed characterization of the fatally injured and, to the extent possible, the tractor involved and the injury event itself. However, additional methods of data collection are needed to provide further characterization of the injury event and the injury agent to design and support prevention programs for tractor users. PMID- 11942562 TI - Characterization of rural mental health service systems. AB - This paper explores two mental health systems in rural North Carolina that provide services to people with severe mental disorders. Recent findings show rural people with mental disorders receive less mental health care than their urban counterparts. This study asks whether rural service systems differ from urban systems in the way that their services are coordinated and structured. A popular conception is that public mental health systems in the United States are uncoordinated with many services provided outside the mental health sector. Rural service providers are seen as even more dependent on nonspecialized mental health providers than their urban counterparts. While many rural service barriers are attributed to the rural environment, little is known about rural service systems and how their organization might contribute to or negate barriers to care. Social network methods were used in this study to compare two rural with four urban systems of care. Findings confirm that mental health systems fit the de facto hypothesis, but that rural systems differ in ways not anticipated by the hypothesis. Rather than being more dependent on nonmental health agencies, rural mental health agencies are more interdependent. PMID- 11942563 TI - Treatment of depression in rural Arkansas: policy implications for improving care. AB - Policy-makers have long suspected that greater barriers to care result in depressed rural residents being less likely to receive high-quality treatment. This study recruited 470 depressed community residents in a 1992 telephone survey, followed 95 percent of them through one year, and abstracted additional data on their health care utilization from insurance claims, medical and pharmacy records. Bivariate and multivariate models demonstrated that during the year following the baseline, there were no significant rural-urban differences in the rate (probability of any outpatient depression treatment), type (probability of receiving general medical depression care only), or quality (completion of guideline-concordant acute-stage care) of outpatient depression treatment. Annual expenditures for outpatient depression treatment were lower for rural subjects compared with their urban counterparts. Rural subjects had 3.05 times the odds of being admitted to a hospital for physical problems and 3.06 times the odds of being admitted to a hospital for mental health problems during the year following baseline compared with urban subjects. Cost-offset analyses demonstrate that every dollar invested in depression treatment was associated with a $2.61 decrease in the cost of treating physical problems in depressed rural residents. Limited insurance coverage and limited availability of services were the most significant barriers to specialty and general medical outpatient treatment for depression in both rural and urban residents. More than 80 percent of depressed residents in both rural and urban areas visited a primary care provider during the year following baseline. The potential cost offset of depression treatment in rural populations plus the improvement in productivity observed in both rural and urban populations indicate that it may be economically possible to improve quality of care for depression without bankrupting an already strained health care budget. PMID- 11942564 TI - A rural mental health research agenda: defining context and setting priorities. AB - This article provides a brief overview of research perspectives on rural mental health services and suggests the importance of building an agenda to bring coherence to studies in this area. The need for sound theory and methodology to guide research is emphasized. The importance of better conceptualization of the rural context as a focus of research is addressed, and 14 propositions concerning issues the authors think will advance rural research are presented. This article is intended to stimulate discussion about a research agenda that will lead to better understanding of rural needs for mental health services as well as more responsive service models. PMID- 11942565 TI - Rural primary care providers' perceptions of their roles in the provision of mental health services: voices from the plains. AB - Much has been written about the deficiencies of primary care providers in their treatment patterns, referral patterns and training for treating people with mental disorders. However, there is a growing realization that, regardless of these shortcomings, primary care providers will continue to be sought out by patients for care of mental disorders, due to patient preference, lack of alternatives or other reasons. Thus, a more recent focus has been on improving the link between primary care providers and mental health specialists. This may include integrated clinics, telecommunication links or simply enhancing the competency of primary care providers through clinical practice guidelines, utilization of screening instruments and greater contact with mental health professionals. Conspicuously absent from most of these studies, commissioned reports and policy papers is the voice of the rural primary care provider. Perhaps due to their heavy practice schedules, little has been written from their perspective. Accordingly, this paper presents the findings from a focus group held in early 1998, of a group of rural primary care providers who practice on the Western plains. It is in this forum that these providers discuss how they perceive their role, their treatment and referral patterns and their feelings and relationships with psychiatrists and the mental health system. PMID- 11942566 TI - Emerging behavioral strategies for the prevention of HIV in rural areas. AB - HIV/AIDS prevention efforts have been concentrated in urban areas, despite increases in HIV in nonmetropolitan areas. This study reviews behavioral prevention programs initiated in rural areas and programs that could be adapted for rural contexts. Outcomes from these interventions demonstrate that preventive interventions at the population, community, targeted populations subgroups, and small group levels can reduce high-risk behavior in rural environments and are cost effective to deliver. PMID- 11942567 TI - Service use of rural and urban Medicaid beneficiaries suffering from depression: the role of supply. AB - Despite the prevalence and consequence of depression in rural areas, the literature on treating depression in rural areas is relatively scarce and inconclusive. The use of mental health services by rural people suffering from depression and the role that supply may play in explaining these differences are not well understood. Understanding these issues for rural Medicaid beneficiaries is important as Medicaid managed carefor physical and behavioral health care is expanded to rural areas. This study compares the mental health service use of rural and urban Medicaid beneficiaries, ages 18 to 64, in Maine suffering from depression and examines what influence mental health and primary care supply have in explaining observed differences. Two models are used to estimate the use of ambulatory mental health services: (1) a logit likelihood estimate of whether a beneficiary uses any outpatient mental health services for depression; (2) an ordinary least squares regression estimating the number of annualized ambulatory mental health care visits among users. Rural beneficiaries suffering from depression have lower utilization than urban beneficiaries. Rural and urban Aid for Families with Dependent Children (AFDC)--and Supplemental Security Income (SSI)--beneficiaries suffering from depression rely more on mental health than on general health care providers to receive ambulatory mental health care. Rural beneficiaries (AFDC and SSI) rely relatively more on general health care providers than urban beneficiaries. Multivariate analysis suggests that mental health supply and patient-level factors, but not primary care supply, account for utilization differences. This article describes the need to better understand factors limiting participation of primary care providers and to study the role of supply across multiple states. PMID- 11942568 TI - An unusual renal angiomyolipoma with morphological lymphangioleiomyomatosis features and coexpression of oestrogen and progesterone receptors. AB - Angiomyolipoma is the most common mesenchymal renal tumour, the clonal origin of which has recently been demonstrated. It is composed of varying amounts of blood vessels, smooth muscle and fat. In this report, we describe a renal angiomyolipoma, which is unusual owing to the presence of a lymphangioleiomyomatosis-like component, occurring in a 41-year-old woman suffering from sporadic lymphangioleiomyomatosis. The diagnosis was based on histopathological and immunohistochemical findings. The tumour consisted of an intimate admixture of two components: one was typical of a classical angiomyolipoma and the other was reminiscent of lymphangioleiomyomatosis. HMB45 positivity was found on 5% of the cells of the angiomyolipoma component. Ten percent of the nuclei of the lymphangioleiomyomatosis and angiomyolipoma components expressed oestrogen receptors and 5% progesterone receptors. This case illustrates a very unusual pattern of a renal angiomyolipoma containing a lymphangioleiomyomatosis-like component. The oestrogen and progesterone immunoreactivity suggests that angiomyolipoma could be hormonally dependent. Therefore, we have emphasised the morphological and immunohistochemical similarities between angiomyolipoma and lymphangioleiomyomatosis. PMID- 11942569 TI - Pilocytic astrocytoma arising in a dermoid cyst of the ovary: a case presentation. PMID- 11942571 TI - P-cadherin expression is associated with high-grade ductal carcinoma in situ of the breast. AB - Cadherins are calcium-dependent cell-cell adhesion glycoproteins, separated into several subclasses with distinct adhesive specificities and tissue distribution, which play an important role in many cellular events. We analyse the expression of E-, N- and P-cadherin in a series of ductal carcinoma in situ (DCIS) of the breast, since this disease represents a heterogeneous group, with different risks of progression to invasive breast carcinoma. We also studied the correlation between cadherin expression and DCIS classification systems, namely the Van Nuys and the Holland et al. classification, this latter based on cytonuclear differentiation and cell polarity. Our results showed that, regardless the classification applied, P-cadherin expression is strongly associated with high histological grade of DCIS (P=0.0047) and lack of estrogen receptors (P=0.0008). The use of Holland et al. classification showed a significant correlation between P-cadherin expression and decreased cell polarity (P=0.01). In conclusion, P cadherin expression seems to be more relevant in DCIS pathogenesis than the altered expression of any other cadherin, including the decrease of E-cadherin expression. PMID- 11942570 TI - Juxta-articular myxoma and intramuscular myxoma are two distinct entities. Activating Gs alpha mutation at Arg 201 codon does not occur in juxta-articular myxoma. AB - Juxta-articular myxoma is a rare myxoid tumor of soft tissue that bears a close histologic resemblance to intramuscular myxoma but is distinguished from the latter by its clinical setting and behavior. Activating missense mutations at the Arg 201 codon of the Gs alpha gene ultimately leading to increased levels of cyclic adenosine monophosphate have been implicated in McCune-Albright syndrome and sporadic fibrous dysplasia of bone. Recently, we have demonstrated that the same Gs alpha mutations occur in intramuscular myxomas associated with fibrous dysplasia of bone (Mazabraud's syndrome) as well as in sporadic intramuscular myxoma. The overlapping histologic appearances of juxta-articular myxoma and intramuscular myxoma prompted us to investigate whether there is a relationship between the two entities. We studied this possibility by looking for Gs alpha mutations in juxta-articular myxoma using polymerase chain reaction (PCR) to amplify appropriate genomic DNA fragments extracted from formalin-fixed, paraffin embedded specimens of five juxta-articular myxomas, followed by single-strand conformation polymorphism analysis. Using these techniques, no aberrant bands were detected in any of the five juxta-articular myxomas, indicating that they lack Gs alpha mutations. Moreover, DNA sequencing of the PCR products of two JAMs showed no abnormalities. We conclude that juxta-articular myxomas, in contrast to intramuscular myxomas, do not involve Arg 201 mutations of the Gs alpha gene, indicating that they represent distinct entities with different underlying molecular mechanisms. PMID- 11942572 TI - Tumor angiogenesis as a prognostic factor in thick cutaneous malignant melanoma. A quantitative morphologic analysis. AB - The prognostic value of the extent of neovascularization in cutaneous melanoma is a highly controversial issue. The aim of the current study was to evaluate whether the morphometric analysis of tumor vascularity may be helpful in predicting the clinical outcome of patients with thick cutaneous melanomas. A series of 15 patients with melanoma (>3 mm in thickness) who did not experience disease progression after long-term follow-up (10 years) and 30 matched controls who underwent recurrence and/or metastases were selected for the study. Microvessels were immunohistochemically stained with anti-CD31 antibody. Several parameters, including vessel number, vascular density, vessel area, equivalent circle diameter, perimeter, shape factor, compactness, and the number of vascular ramifications per 100 vessel sections, were quantitatively assessed by a computer aided semi-automatic image analysis system. Mean vessel area was 341.69 microm2 in cases without progression and 512.55 microm2 in the progressed melanomas (P=0.008, Mann-Whitney U test). The mean equivalent circle diameter was 18.95 microm in non-progressed melanomas and 22.57 microm in progressed melanomas (P=0.009). The mean number of ramifications was 0.8 in cases without progression and 1.9 in the controls (P=0.03). Microvessel count and vascular density were higher in progressed cases (17.37 vs. 11.73 and 28.94/mm2 vs. 19.55/mm2, respectively), but the difference did not reach statistical significance (P=0.06). Our results suggest that neovascularization is a critical event in the progression of thick melanoma. Its prognostic significance is better assessed by quantification of vessel area, equivalent circle diameter, and microvessel branching, whereas microvessel count and vascular density do not provide significant prognostic information. PMID- 11942573 TI - Sclerosing polycystic adenosis of parotid gland with dysplasia and ductal carcinoma in situ. Report of three cases with immunohistochemical and ultrastructural examination. AB - We describe three cases of sclerosing polycystic adenosis (SPA) of the parotid gland, a salivary condition analogous to fibrocystic disease of the breast. For the first time, immunoreactivity for oestrogen and progesterone receptors was demonstrated, suggesting a possible participation of hormone stimulation in its pathogenesis. In addition, all our cases showed foci of dysplasia of the ductal epithelium, which in one case was severe enough to amount to carcinoma in situ. This feature that has not previously been reported in SPA. PMID- 11942574 TI - Preneoplastic non-papillary lesions and conditions of the urinary bladder: an update based on the Ancona International Consultation. AB - BACKGROUND AND AIMS: This paper summarizes the work done by the members of the Committee no. 2 at the International Consultation on the Diagnosis of Non Invasive Urothelial Neoplasms held in Ancona, Italy (11-12 May 2001). The committee members discussed and reached consensus regarding the optimal contemporary diagnosis and classification of the preneoplastic non-papillary lesions of the urothelium. An important objective was to promote a precise terminology and to use it consistently in daily practice in pathology and urology. RESULTS AND CONCLUSIONS: The result of the meeting is represented by a refined classification of the non-papillary intraepithelial lesions and conditions of the urothelium. This classification includes epithelial abnormalities (reactive urothelial atypia and flat urothelial hyperplasia), presumed preneoplastic lesions and conditions (keratinizing squamous and glandular metaplasia, and malignancy-associated cellular changes), as well as preneoplastic (dysplasia) and neoplastic non-invasive (carcinoma in situ) lesions. Each of these lesions is defined with strict morphological criteria in order to provide more accurate information to urologists in managing patients. PMID- 11942575 TI - Spindle-cell squamous carcinoma exhibiting myofibroblastic differentiation. A study of two cases showing fibronexus junctions. AB - The features of two spindle-cell carcinomas of the dermis are described with special reference to the presence of fibronexus junctions in tumour cells. The cases were of a 78-year-old man with a left eye-lid tumour and a 78-year-old woman with a naso-labial fold tumour, who had been given radiotherapy 13 years earlier for a clinically diagnosed basal cell carcinoma. Both specimens were slightly ulcerated and polypoid. Histologically, invasive tumour consisted of interlacing fascicles of plump spindled and oval cells, which were positive for several anti-cytokeratin antibodies, epithelial membrane antigen, vimentin and smooth-muscle and muscle-specific actins. Ultrastructurally, tumour cells in both cases contained rough endoplasmic reticulum (prominent in case 1), tonofibrils, desmosomes and smooth-muscle type myofilaments with focal densities. Fibronexus junctions were also present, which correlated with positive staining for fibronectin. This is the first documentation of the fibronexus in epithelial tissue in vivo, and the first unambiguous demonstration in a spindle-cell carcinoma. It extends the known distribution of the fibronexus and expands the ultrastructural spectrum of squamous carcinoma. PMID- 11942576 TI - DNA content and expression of cell cycle proteins in caterpillar nuclei from fetal human cardiac myocytes. AB - Caterpillar nuclei (CN) are characterized by their peculiar morphology, with chromatin distributed in clusters and running along the longitudinal axis of the nucleus. They can be observed in normal hearts of fetuses as well as in hearts of children and adults with rheumatic heart disease. This study has demonstrated by means of ploidy studies with digital image analysis that in the fetal heart (20.5+/-1.8 weeks) the CN (diploid = 5.6+/-8.4%; tetraploid = 46.2+/-24.2%; hypertetraploid = 46.9+/-26.3%) present higher DNA content than non-caterpillar myocyte nuclei (diploid = 89.4+/-6.2%; tetraploid = 10.0+/-4.1%; hypertetraploid = 1.5+/-1.3%) (P=0.000001, 0.00013, and 0.000038, respectively). Expression of proliferation cell nuclear antigen (PCNA; 30.6+/-11.7% in CN and 13.4+/-7.3% in non-caterpillar myocyte nuclei; P=0.0115) and cyclin B1 (2.8+/-3.8% and 12.6+/ 15.6%, respectively; P=n.s.) was also positive in these nuclei. In conclusion, these results suggest that there exists a relationship between CN morphology and myocyte replication phenomena. PMID- 11942577 TI - Ultraviolet light-irradiated collagen III modulates expression of cytoskeletal and surface adhesion molecules in rat aortic smooth muscle cells in vitro. AB - Systemic and pulmonary hypertension is characterised by structural reconstruction of the vascular wall which includes hypertrophy and hyperplasia of vascular smooth muscle cells (VSMCs) and fibroproduction. We hypothesise that these changes are stimulated by non-enzymatic modification of collagen molecules in the injured vascular wall by radicals. We exposed collagen III to ultraviolet (UV) light irradiation which, as indicated by fluorescence and electrophoretic analyses, resulted in its fragmentation. Both irradiated and control unmodified collagen were adsorbed on culture dishes and seeded with VSMCs derived from the rat thoracic aorta. During the first week after seeding, the cells on the modified collagen attained significantly higher population density (by 15-83%), higher mitotic index (by 31-135%) and higher BrdU labelling index (by 32%). However, these cells were less resistant to spontaneous and trypsin-mediated detachment from the growth support. As revealed using enzyme-linked immunosorbent assay in 3-day-old cultures, the cells growing on the irradiated collagen exhibited a lower concentration of beta-1 integrins (-10%, measured per milligram of protein), vinculin (-18%), talin (-6%) and vimentin (-15%). Immunofluorescence staining showed that these molecules were distributed more diffusely and less organised into focal adhesion plaques or cytoskeletal fibres. The concentration of two adhesion molecules of immunoglobulin type, ICAM-1 and VCAM-1, was increased by 11% and 16%, respectively. The concentration of alpha-v integrins and alpha-actin was unchanged; the latter, however, formed fewer distinct microfilament bundles in cells on the modified collagen. Our results suggest that the VSMCs growing on UV-modified collagen are more prone to escape the growth control mediated by cell-extracellular matrix contact and can bind the cells of the immune system. PMID- 11942578 TI - Gradual loss of pancreatic beta-cell insulin, glucokinase and GLUT2 glucose transporter immunoreactivities during the time course of nutritionally induced type-2 diabetes in Psammomys obesus (sand rat). AB - The Psammomys obesus (sand rat) is a well-established model of nutritionally induced non-insulin-dependent type-2 diabetes. When fed a high-energy (HE) diet, the diabetes-prone animals develop hyperinsulinaemia and hyperglycaemia. Within 1 week, all animals become hyperinsulinaemic. However, a loss of immunostaining for insulin as well as for the GLUT2 glucose transporter in the plasma membrane and the glucokinase in the cytoplasm of the pancreatic beta cells became evident only when the animals subsequently developed hyperglycaemia. After 1 week of HE diet feeding, the pancreatic beta-cell volume was reduced by one-third in hyperglycaemic Psammomys. Insulin immunostaining as well as GLUT2 glucose transporter immunostaining in the plasma membrane and glucokinase immunostaining in the cytoplasm were reduced by more than 50%. After 3 weeks of HE diet feeding, all changes observed after 1 week were even more pronounced, with reductions in the range of 70-95%. The reduction of the total beta-cell volume of the pancreas due to beta-cell death and the diminution of insulin content of the remaining beta cells in the islets during the HE diet feeding was accompanied by a parallel fall of the pancreas insulin content. For all changes observed, there was a significant correlation with the increase of the blood glucose concentration (r>0.9) but not with the increase of the plasma insulin concentration (r>0.2). Thus, increasing glycaemia appears to be the factor responsible for the deterioration of the pancreatic beta-cell function and the resulting loss of the insulin secretory capacity in Psammomys. The final result of this development is an irreversible diabetic state due to the feeding of the HE diet. PMID- 11942579 TI - Cytokeratin typing as an aid in the differential diagnosis of primary versus metastatic lung carcinomas, and comparison with normal lung. AB - Due to more efficient chemotherapy protocols, the number of second and even third primary carcinomas is steadily increasing. To denominate the possible origin of a carcinoma, different markers are available as an aid, e.g. hormones, proteins and lipoproteins, secretion products and cytoskeletal proteins. Cytokeratins (CKs) have gained new popularity; however, they have not been extensively evaluated in lung tumours. In our study we evaluated the staining patterns of CK polypeptides 4-8, 10, 13, 14, and 17-20 and high molecular weight (HMW) CK polypeptides in routinely processed primary lung carcinomas and lung metastases of diverse origin. As expected, immunohistochemical investigation gave no clear-cut results, but, with statistical analysis, lung adenocarcinomas could be separated from metastatic adenocarcinomas using CK 5 and 18 and HMW CK (specificity 92.5%, sensitivity 62.5%). The different origin of the metastases could often be detected using CK 18 and CK 20. Lung clear cell carcinomas and large cell carcinomas with clear cell areas could be distinguished from metastatic renal clear cell carcinomas by the CK 7 staining reaction. Squamous cell carcinomas of the lung and metastatic squamous cell carcinomas of the larynx, pharynx and oesophagus could not reliably be separated in part due to the few number of cases available. CK polypeptide typing is thus an additional aid in the differential diagnosis of lung carcinomas versus carcinomas metastatic to the lung. PMID- 11942580 TI - Loss of heterozygosity at chromosome 6q23-25 correlates with clinical and histologic parameters in salivary gland adenoid cystic carcinoma. AB - The prognosis of salivary gland adenoid cystic carcinoma (ACC) depends on the clinical stage, the location of the primary tumor, and the histologic growth pattern. ACCs with a cribriform growth pattern have a better prognosis than those with a solid growth pattern; however, clear-cut grading criteria have not yet been established, and therefore prognostic indicators on a molecular level are of special interest. In order to analyze tumor tissue with different growth patterns, cribriform and solid tumor areas of 25 patients were microdissected and separately analyzed for loss of heterozygosity (LOH) at nine polymorphic microsatellite markers located between 6q14 and 6q27. LOH was detected in 19/25 (76%) patients and LOH rates were highest at markers D6S441, D6S310, D6S311 and UTRN, which are located at 6q23-25. Combined analysis of LOH at these four markers shows that in primary tumor subtype foci with cribriform growth pattern LOH is associated with high TNM stages (P<0.01), high T stages (P=0.01), positive lymph node status (P=0.03), an unfavorable disease course (P=0.02), and the presence of >10% solid growth pattern (P=0.05). In contrast, in primary tumor subtype foci with solid growth pattern, no significant differences in LOH rates were found in patients from prognostically and histologically favorable versus unfavorable patient groups. The frequent occurrence of LOH at 6q23-25 and the correlation of LOH rates with prognostic parameters indicate that a prognostically important tumor suppressor gene is located in this chromosomal area. PMID- 11942581 TI - Genetic evidence for the multi-step progression of mixed glandular-neuroendocrine gastric carcinomas. AB - A mixed glandular-neuroendocrine gastric carcinoma shows discrete, juxtaposed areas of adenocarcinoma and neuroendocrine carcinoma. In order to gain insight into the genetic events and clonality associated with the dual differentiation of a mixed gastric carcinoma, eight cases (two true composite and six neuroendocrine dominant carcinomas) were examined by analyzing the genome-wide loss of heterozygosity (LOH). Of the eight mixed gastric carcinomas, one true composite and five neuroendocrine-dominant carcinomas showed a primary LOH that was shared by both the glandular and neuroendocrine components and a secondary LOH or mutations that were restricted to the neuroendocrine components. The glandular components contained mixed cell populations with or without primary LOH events, suggesting that a primary LOH arose during adenocarcinoma progression and that the LOH-positive cell served as a precursor for a neuroendocrine carcinoma. The neuroendocrine components were of a homogeneous population with various genetic alterations such as a LOH, p53 mutations, and microsatellite instability associated transforming growth factor (TGF)-beta RII mutation. Therefore, most (six of eight cases) mixed glandular-neuroendocrine gastric carcinomas were likely to have sequentially evolved from a glandular precursor to a genetically heterogeneous adenocarcinoma and then to neuroendocrine differentiation. Two components of the other true composite carcinomas were shown to have reciprocally lost different alleles of identical loci on multiple chromosomes, suggesting that, occasionally (one of eight cases), dual differentiation concurrently arises from a single precursor, possibly as a result of non-disjunctional cell division. PMID- 11942582 TI - Development of renal structural lesions in type-1 diabetic patients with microalbuminuria. Observations by light microscopy in 8-year follow-up biopsies. AB - Kidney biopsies were obtained in 18 type-1 diabetic patients with microalbuminuria and again 8 years later. Over the first 30 months, a treatment protocol (conventional versus intensified treatment) was followed. The biopsies were embedded into plastic and sectioned serially. The volume of individual glomeruli and the vascular pole area was determined. The number of glomeruli showing capsular drops, fibrinoid lesions, adhesions, extra efferent arterioles and corpuscles totally occluded were counted and expressed as a percentage of the total number of corpuscles. Cortical interstitium and degenerated tubules were estimated by point counting. From baseline to the 8-year biopsy, the group of patients showed significant increase in interstitial volume fraction, mean glomerular volume and vascular pole area. The frequency of glomerular occlusion, fibrinoid lesions, adherences and extra efferent arterioles increased, whereas the frequency of capsular drops did not change, and degenerated tubular profiles showed a non-significant increase. No correlation was seen between these light microscopic observations and the concomitant development of diabetic glomerulopathy. Increase in albumin excretion rate was associated with increase in glomerular volume and vascular pole area. No correlation was seen with the glomerular filtration rate, blood pressure or metabolic control. The increase in structural parameters illustrates the slowly ongoing alteration of renal structures in type-1 diabetic patients with microalbuminuria. PMID- 11942583 TI - Atrioventricular nodal reentrant tachycardia in children: effect of slow pathway ablation on fast pathway function. AB - INTRODUCTION: Prior studies in adults have shown significant shortening of the fast pathway effective refractory period after successful slow pathway ablation. As differences between adults and children exist in other characteristics of AV nodal reentrant tachycardia (AVNRT), we sought to characterize the effect of slow pathway ablation or modification in a multicenter study of pediatric patients. METHODS AND RESULTS: Data from procedures in pediatric patients were gathered retrospectively from five institutions. Entry criteria were age <21 years, typical AVNRT inducible with/without isoproterenol infusion, and attempted slow pathway ablation or modification. Dual AV nodal pathways were defined as those with > or =50 msec jump in A2-H2 with a 10-msec decrease in A1-A2. Successful ablation was defined as elimination of AVNRT inducibility. A total of 159 patients (age 4.4 to 21 years, mean 13.1) were studied and had attempted slow pathway ablation. AVNRT was inducible in the baseline state in 74 (47%) of 159 patients and with isoproterenol in the remainder. Dual AV nodal pathways were noted in 98 (62%) of 159 patients in the baseline state. Ablation was successful in 154 (97%) of 159 patients. In patients with dual AV nodal pathways and successful slow pathway ablation, the mean fast pathway effective refractory period was 343+/-68 msec before ablation and 263+/-64 msec after ablation. Mean decrease in the fast pathway effective refractory period was 81+/-82 msec (P < 0.0001) and was not explained by changes in autonomic tone, as measured by changes in sinus cycle length during the ablation procedure. Electrophysiologic measurements were correlated with age. Fast pathway effective refractory period was related to age both before (P = 0.0044) and after ablation (P < 0.0001). AV block cycle length was related to age both before (P = 0.0005) and after ablation (P < 0.0001). However, in dual AV nodal pathway patients, the magnitude of change in the fast pathway effective refractory period after ablation was not related to age. CONCLUSION: Lack of clear dual AV node physiology is common in pediatric patients with inducible AVNRT (38%). Fast pathway effective refractory period shortens substantially in response to slow pathway ablation. The magnitude of change is large compared with adult reports and is not completely explained by changes in autonomic tone. Prospective studies in children using autonomic blockade are needed. PMID- 11942584 TI - Influences on fast and slow pathway conduction in children: does the definition of dual atrioventricular node physiology need to be changed? AB - The present study clearly establishes that there are a variety of influences on fast and slow pathway conduction properties and that there are subpopulations of children in whom these conduction properties differ. A more detailed comparison of data from these subpopulations may provide more information regarding the relative strength of these influences and how they change. In turn, this may lead to a more accurate and useful definition of dual AV node physiology in children and an improved understanding of the etiology of AVNRT. PMID- 11942585 TI - Pronounced effect of procainamide on clockwise right atrial isthmus conduction compared with counterclockwise conduction: possible mechanism of the greater incidence of common atrial flutter during antiarrhythmic therapy. AB - INTRODUCTION: It has been shown that the induction rates of common and reversed common atrial flutter are comparable during baseline control study, whereas the rate is significantly greater for common flutter than reversed common flutter during administration of antiarrhythmic agents. The mechanism of this discrepancy is not known. METHODS AND RESULTS: The study consisted of 15 patients (group 1) with clinically documented common atrial flutter either with (n = 10) or without (n = 5) Class I antiarrhythmic therapy, and 15 patients with paroxysmal supraventricular tachycardia (group 2). Bidirectional conduction velocity and minimal pacing cycle length of 1:1 conduction during incremental pacing from both the low lateral right atrium and coronary sinus were assessed. The response of these variables to procainamide was analyzed in correlation with the induction rate of each type of flutter during the pacing protocol. Conduction velocity in the clockwise (CW) direction was significantly slower for all pacing cycle lengths than conduction velocity in the counterclockwise (CCW) direction in group 1 but was similar in group 2. Minimal pacing cycle length of 1:1 conduction did not differ between CW and CCW conduction in either group. However, in group 1, minimal pacing cycle length of 1:1 conduction of CW conduction was prolonged to a greater degree after procainamide than that of CCW conduction. There also was a significant increase in the induction rate of common flutter. This preferential effect of procainamide on CW conduction was not observed in group 2. CONCLUSION: CW conduction over the isthmus is preferentially influenced by procainamide compared with CCW conduction, which may explain the greater incidence and induction probability of common flutter during antiarrhythmic therapy. PMID- 11942586 TI - Mechanisms for atrial fibrillation in patients with Wolff-Parkinson-White syndrome. AB - INTRODUCTION: Paroxysmal atrial fibrillation (PAF) frequently occurs in patients with Wolff-Parkinson-White (WPW) syndrome. To elucidate the mechanisms for PAF, we performed electrophysiologic studies (EPS) before and after ablation of accessory pathways (APs). METHODS AND RESULTS: We investigated 24 patients with WPW syndrome who had AV reciprocating tachycardia and prior PAF and had undergone successful ablation of APs. Patients in whom atrial fibrillation (AF) was induced by EPS at day 7 after ablation were considered the inducible AF group (n = 14), and patients in whom AF was not induced by EPS at day 7 after ablation were considered the noninducible AF group (n = 10). Fifteen patients with AV nodal reentrant tachycardia (AVNRT) but without PAF who underwent ablation of the slow AV nodal pathways served as the control group (AVNRT group). Maximal atrial conduction delay and conduction delay zone, which are indices of atrial vulnerability, were measured before and after ablation. Before ablation, maximal atrial conduction delay and conduction delay zone were significantly greater (P < 0.0001 and P < 0.0001, respectively) in the two WPW syndrome groups than in the AVNRT group, indicating increased atrial vulnerability in WPW syndrome with PAF. After ablation, these parameters did not change in the inducible AF group, whereas they were significantly (P < 0.0001) decreased in the noninducible AF group and were not different from those in the AVNRT group, indicating normalized atrial vulnerability in the noninducible AF group after ablation. The prospective study demonstrated that PAF recurred only in the inducible AF group during long term follow-up (17+/-7 months). CONCLUSION: The findings of this study suggest that there are two mechanisms of PAF in patients with WPW syndrome: one mechanism is reversible and AP-dependent atrial vulnerability, and the other is intrinsic and AP-independent atrial vulnerability. PMID- 11942587 TI - Atrial fibrillation in patients with Wolff-Parkinson-White syndrome. PMID- 11942588 TI - Risk of sudden death after successful accessory atrioventricular pathway ablation in resuscitated patients with Wolff-Parkinson-White syndrome. AB - INTRODUCTION: Cardiac arrest in patients with Wolff-Parkinson-White (WPW) syndrome can be due to ventricular fibrillation mediated by fast conduction over the accessory pathway during atrial fibrillation. However, if primary ventricular fibrillation is the reason for resuscitation, placement of an implantable cardioverter defibrillator (ICD) would be indicated. The aim of this study was to test the hypothesis that in resuscitated patients with WPW syndrome, recurrences can be prevented by sole ablation of their accessory pathways. METHODS AND RESULTS: We performed a long-term follow-up study of 48 resuscitated patients with WPW syndrome who underwent successful accessory pathway ablation as their sole primary treatment. Cardiac arrest had occurred either spontaneously in 32 patients (group A) or after intravenous administration of antiarrhythmic drugs in 16 patients (group B) and was never associated with an acute myocardial infarction or other concomitant factors. All patients had normal left ventricular function at echocardiography. A total of 56 accessory AV pathways were ablated successfully with radiofrequency current (n = 55) or during surgery (n = 1) and were located at the left free wall (n = 35), right free wall (n = 8), or septal paraseptal region (n = 13). Follow-up 5.0+/-1.9 years after ablation (range 0.2 to 7.9) was obtained in all 48 patients. All of the patients were alive, and none had a life-threatening arrhythmia or syncope after successful ablation of their accessory pathways. CONCLUSION: In resuscitated patients with WPW syndrome who have normal left ventricular function at echocardiography and no ECG abnormalities suggesting additional electrical disease, ablation of their overt accessory pathways prevented cardiac arrest recurrences; therefore, ICD placement is generally not indicated. PMID- 11942589 TI - Prevalence of a shared isthmus in postinfarction patients with pleiomorphic, hemodynamically tolerated ventricular tachycardias. AB - INTRODUCTION: Multiple forms of ventricular tachycardia (VT) after myocardial infarction may result from multiple reentrant circuits that share an isthmus or from separate reentrant circuits. The prevalence of a shared isthmus in patients with multiple hemodynamically tolerated VTs has not been determined. METHODS AND RESULTS: Criteria for a shared isthmus consisted of (1) concealed entrainment of >1 VT at a single pacing site; (2) concealed entrainment during VT and a perfect pace map of another VT at the same pacing site; or (3) concealed entrainment of VT of a given morphology that had at least two cycle lengths that varied by at least 100 msec. In a series of 19 patients (16 men and 3 women; age 65+/-14 years, ejection fraction 0.25+/-0.09) with 54 VTs (mean cycle length 494+/-98 msec), there was evidence of a shared isthmus in 23 VTs (43%) at 11 sites in 9 patients. Concealed entrainment of two different VTs was observed at 4 of 11 sites. At 5 of 11 sites there was concealed entrainment of one VT and a perfect pace map of another VT. At the remaining 2 of 11 sites, there was concealed entrainment of a VT that had two different cycle lengths. Nineteen of the 23 VTs were ablated successfully with radiofrequency energy applications at 11 sites. CONCLUSION: In postinfarction patients with pleiomorphic, hemodynamically stable VT, a shared isthmus may be present in approximately 40% of VTs. PMID- 11942590 TI - Two for the price of one: identifying shared circuits in postinfarction reentrant ventricular tachycardia. PMID- 11942591 TI - Appraisal of "Mahaim" automatic tachycardia. AB - A series of four patients with right-sided accessory pathways with long conducting times and decremental properties is reported. All patients underwent radiofrequency catheter ablation, and target areas were guided by a discrete "Mahaim" potential recorded at the lateral aspect of the tricuspid valve. A slow automatic and irregular rhythm with a QRS morphology similar to that of a fully preexcited QRS complex occurred during radiofrequency current delivery. The occurrence of so-called "Mahaim" automatic tachycardia heralded successful elimination of the accessory pathway in a manner similar to that of junctional automatic rhythm during slow pathway ablation in patients with AV nodal reentrant tachycardia. The observation of an automatic rhythm brought about during radiofrequency current ablation of a Mahaim-like accessory pathway is electrophysiologic evidence of the accessory AV nodal behavior of this structure. PMID- 11942592 TI - Response of atriofascicular pathways to radiofrequency ablation: further evidence of an ectopic atrioventricular node? PMID- 11942593 TI - Polymorphisms in beta-adrenergic receptor genes in the acquired long QT syndrome. AB - INTRODUCTION: Sympathetic activation is a trigger for life-threatening arrhythmias in many patients with the congenital long QT syndrome (LQTS), and an increase in heart rate has been reported just prior to torsades de pointes in patients with drug-associated (acquired) LQTS (aLQTS). We compared the frequencies of five recognized nonsynonymous coding region polymorphisms in genes encoding the beta1-adrenergic and beta2-adrenergic receptors (AR) in 93 patients with aLQTS and 3 control groups: an ethically diverse set of individuals from middle Tennessee (n = 71), a subset of the Polymorphism Discovery Resource obtained from National Human Genome Research Institute (n = 89), and patients who tolerated QT-prolonging drugs without aLQTS (non-aLQTS group; n = 66). METHODS AND RESULTS: Polymerase chain reaction-restriction fragment length polymorphism was used to screen for Ser49Gly and Gly389Arg (beta1-AR) and Thr164Ile (beta2 AR). For Arg16Gly and Gln27Glu, polymorphic sites 33 nucleotides apart in the beta2-AR, single-stranded conformational polymorphism was used to distinguish among the 4 possible haplotypes and 10 possible genotypes. Allele frequencies were similar among the 4 groups at the 2 beta1-AR sites. The uncommon Ile164 variant in beta2-AR was slightly more frequent in patients (3.2%) than in any of the 3 control groups (0.6% to 2.3%). At the 16-27 neighboring sites in the beta2 AR, one haplotype (Arg16/Glu27) was not detected, as in previous studies; hence, only 6 genotypes were present. There were fewer Gly16/Gln27 homozygotes in the non-aLQTS group (1.5%) than in two other control groups or the aLQTS group (8.5% to 10%). CONCLUSION: None of the five common nonsynonymous coding region polymorphisms in the beta-AR genes predict drug-associated torsades de pointes, although the Gly16/Gln27 haplotype may be a risk factor. PMID- 11942594 TI - Histopathologic exploration of intra-hisian conduction disturbances. AB - INTRODUCTION: The length of the His bundle and the precise location of injury responsible for split His potentials have not been fully established in patients with intra-Hisian block. We conducted an autopsy study comparing histologic findings in intra-Hisian block versus control hearts. METHODS AND RESULTS: We studied hearts from 4 intra-Hisian block patients (age 66 to 93 years, mean 79.5) and hearts from 14 patients without AV conduction abnormalities (control). All intra-Hisian block patients underwent electrophysiologic evaluation; 3 patients demonstrated intra-Hisian block and 1 showed no His potential. Autopsies were performed when each patient died. After the heart was fixed in formaldehyde, the AV septal junctional area was removed en bloc and serially sectioned into 7 microm thick slices. For study purposes, we considered the three segments of the His bundle separately: the penetrating bundle, the nonbranching bundle, and the branching bundle. The actual length of each segment was calculated from the number of respective serial sections, and the lesion was reconstructed within the conduction axis. Intra-Hisian block hearts were heavier than control hearts (mean weight 389 vs 301 g; P < 0.05). The lesion was situated in the nonbranching bundle in 3 hearts and in the penetrating bundle in 1 heart. Mean compact node length was 3.8 mm in intra-Hisian block hearts and 3.3 mm in control hearts. The penetrating bundle was 2.1 and 2.1 mm, the nonbranching bundle was 3.5 and 1.9 mm, and the branching bundle was 4.5 and 4.6 mm in intra-Hisian block and control hearts, respectively. CONCLUSION: Most lesions were observed in the nonbranching bundle adjacent to the junction between the central fibrous body and ventricular septum. This segment was longer in intra-Hisian block hearts than in control hearts. PMID- 11942595 TI - Pathologic basis of conduction disturbances: the challenge to move beyond microscopic anatomy. PMID- 11942596 TI - Supervulnerable phase immediately after termination of atrial fibrillation. AB - INTRODUCTION: Recent studies with the implantable atrial cardioverter have shown that atrial fibrillation (AF) recurs almost immediately after successful cardioversion in about 27% of cases. In the present study, we determined the electrophysiologic properties of the caprine atrium immediately after spontaneous termination of AF both before and after 48 hours of AF-induced electrical remodeling. METHODS AND RESULTS: In eight goats, atrial effective refractory period (AERP), intra-atrial conduction velocity, and atrial wavelength were measured during sinus rhythm both before (t = 0) and after 48 hours (t = 48) of electrically maintained AF (baseline). After baseline, a 5-minute paroxysm of AF was induced, during which the refractory period (RPAF) was determined. AERP, conduction velocity, and atrial wavelength also were measured immediately after spontaneous restoration of sinus rhythm (post-AF values). Both in normal and remodeled atria, immediately after AF, AERP and conduction velocity were markedly decreased compared with baseline (P < 0.01). In normal atria, post-AF AERP (107+/ 14 msec) gradually prolonged from its AF value (114+/-17 msec) to its baseline value (138+/-13 msec). Conduction velocity decreased from 130+/-9 cm/sec to 117+/ 9 cm/sec. After 48 hours of AF, AERP had shortened to 74+/-8 msec. RPAF was 89+/ 9 msec. Surprisingly, immediately after termination of AF, AERP shortened further to 58+/-6 msec (P < 0.01). Post-AF conduction velocity decreased from 136+/-11 cm/sec to 122+/-10 cm/sec (P < 0.01). As a result, the post-AF atrial wavelength became as short as 7.1+/-1 cm. These changes were transient, and all parameters gradually returned to baseline within 1 to 2 minutes after conversion of AF. CONCLUSION: Due to a combined decrease in AERP and conduction velocity, marked shortening of the atrial wavelength occurs during the first minutes after conversion of AF. In electrically remodeled atria, this results in a transient ultrashort value of AERP (<60 msec) and atrial wavelength (7.1 cm). These observations imply a highly vulnerable substrate for reentry immediately after termination of AF. During this supervulnerable phase, both early and later premature beats reinitiated immediate recurrences of AF. PMID- 11942597 TI - Effects of estrogen on cardiac electrophysiology in female mice. AB - INTRODUCTION: Understanding the molecular mechanisms that underlie gender- and hormonal-related differences in susceptibility to cardiac arrhythmias has been hampered by the lack of a suitable animal model. We examined the effect of hormonal status on the electrophysiologic (EP) properties of the mouse heart in an in vivo, closed chest model. METHODS AND RESULTS: Fifty-three female C57/J mice aged 10 to 12 weeks were studied. Thirty-six mice underwent bilateral ovariectomies; 18 received estrogen (OVX+E) and 18 received placebo (OVX). Seventeen female mice underwent only sham surgery. All animals underwent in vivo EP studies. Select EP parameters were measured after quinidine treatment. Data were analyzed by a blinded observer. Compared with the intact female mice, the PR and AH intervals were significantly shorter in the OVX mice, and these parameters normalized with estrogen replacement (PR = 45.9+/-4.5 msec in the intact mice, 42.1+/-4.3 msec in the OVX group, and 46.9+/-3.5 msec in the OVX+E group, P < 0.005; AH = 36.5+/-4.9 msec in the intact mice, 34.4+/-4.7 msec in the OVX group, and 38.8+/-2.7 msec in the OVX+E group, P = 0.03). The right ventricular effective refractory period was significantly shorter in the OVX mice versus the intact mice, and this also normalized with estrogen replacement. Hormonal status did not significantly affect any other EP variable, including QT interval. CONCLUSION: In female mice, estrogen prolongs AV nodal conduction and the right ventricular effective refractory period. Taken together, these data suggest that hormonal status affects aspects of cardiac EP function. Future application of this mouse model will be helpful in determining the molecular pathways that mediate hormonal differences in cardiac EP. PMID- 11942598 TI - Radiofrequency catheter ablation of idiopathic ventricular tachycardia originating in the main stem of the pulmonary artery. AB - We report the case of a patient in whom successful radiofrequency catheter ablation of an idiopathic ventricular tachycardia (VT) originating in the main stem of the pulmonary artery was performed. After successful ablation of the index arrhythmia, which was an idiopathic right ventricular outflow tract VT, a second VT with a different QRS morphology was reproducibly induced. Mapping of the second VT revealed the presence of myocardium approximately 2 cm above the pulmonary valve. Application of radiofrequency energy at this site resulted in termination and noninducibility of this VT. After 6-month follow-up, the patient remained free from VT recurrences. PMID- 11942599 TI - Accessory pathway automaticity after radiofrequency ablation. AB - Mapping in a patient undergoing radiofrequency ablation for a left-sided concealed accessory pathway showed that the site with the shortest VA conduction time was in the great cardiac vein. Epicardial radiofrequency delivery at that site was successful. After ablation, a potential dissociated from both atrial and ventricular activity during sinus rhythm and atrial pacing was noted on the distal bipole of the ablation catheter. During incremental ventricular pacing, intermittent ventricular to potential conduction was observed. This indicates complete block at the atrial-accessory pathway interface and impaired conduction at the ventricular interface. This dissociated activity originating neither from the ventricle nor from the atria provides evidence of automatic accessory pathway activity. PMID- 11942600 TI - Automaticity in the coronary sinus. PMID- 11942601 TI - Embolization of pacing electrode fragment into the superolateral vein in the spinal canal causing root compression. AB - Lead extraction may be complicated by lead fracture and embolization of the lead fragments into the pulmonary circulation or other vasculature causing various signs and symptoms. We present a very unusual case demonstrating embolization of a pacemaker electrode fragment into the superolateral vein in the spinal canal causing root compression. The electrode fragment was removed by spinal surgery, and the patient's symptoms gradually resolved. PMID- 11942602 TI - Electrical restitution and cardiac fibrillation. AB - Combined experimental and theoretical work has shown that restitution properties of the cardiac action potential duration and conduction velocity contribute to breakup of reentrant wavefronts during cardiac fibrillation independent of preexisting electrophysiologic heterogeneities in the tissue. Developing therapies that favorably alter these cardiac electrical restitution properties are a promising new approach to preventing fibrillation. PMID- 11942603 TI - Atrial pacing during a narrow QRS complex tachycardia: what is the mechanism? PMID- 11942604 TI - A cool ablation. PMID- 11942605 TI - Adenosine triphosphate enhanced contrast pulmonary venogram to facilitate pulmonary vein ablation. PMID- 11942606 TI - Cytokinesis in budding yeast: the relationship between actomyosin ring function and septum formation. AB - Cytokinesis in budding yeast is accomplished by the concerted action of actomyosin ring function and septum formation. The actomyosin ring is not essential for cell viability, but it is required for efficient cell division. Deletion of the actomyosin ring results in abnormal septum formation, and a delay in cytokinesis and cell separation. In contrast, septum formation is essential for cell viability. Block of septum formation prevents the contraction, but not the formation of the actomyosin ring. Here we review and provide additional evidence that defines the functional and molecular relationship between actomyosin ring function and septum formation. PMID- 11942607 TI - Studies in fission yeast on mechanisms of cell division site placement. AB - One fundamental problem in cytokinesis is how the plane of cell division is established. In this review, we describe our studies on searching for "signals" that position the cell division plane, using fission yeast Schizosaccharomyces pombe. First, we take a genetic approach to determine how the nucleus may position the contractile ring in fission yeast. mid1p appears to link the position of the ring with the nuclear position, as it is required for proper placement of the contractile ring and is localized in a band at the cell surface overlying the nucleus. Second, we study how microtubules may function in the establishment of cell polarity at the cell tips. tea1p may be deposited on the cell surface by microtubules and function to recruit proteins involved in making actin structures. These studies suggest how microtubules may direct the assembly of the contractile ring in animal cells. PMID- 11942608 TI - Contractile ring formation in Xenopus egg and fission yeast. AB - How actin filaments (F-actin) and myosin II (myosin) assemble to form the contractile ring was investigated with fission yeast and Xenopus egg. In fission yeast cells, an aster-like structure composed of F-actin cables is formed at the medial cortex of the cell during prophase to metaphase, and a single F-actin cable(s) extends from this structure, which seems to be a structural basis of the contractile ring. In early mitosis, myosin localizes as dots in the medial cortex independently of F-actin. Then they fuse with each other and are packed into a thin contractile ring. At the growing ends of the cleavage furrow of Xenopus eggs, F-actin at first assembles to form patches. Next they fuse with each other to form short F-actin bundles. The short bundles then form long bundles. Myosin seems to be transported by the cortical movement to the growing end and assembles there as spots earlier than F-actin. Actin polymerization into the patches is likely to occur after accumulation of myosin. The myosin spots and the F-actin patches are simultaneously reorganized to form the contractile ring bundles. The idea that a Ca signal triggers cleavage furrow formation was tested with Xenopus eggs during the first cleavage. We could not detect any Ca signals such as a Ca wave, Ca puffs or even Ca blips at the growing end of the cleavage furrow. Furthermore, cleavages are not affected by Ca-chelators injected into the eggs at concentrations sufficient to suppress the Ca waves. Thus we conclude that formation of the contractile ring is not induced by a Ca signal at the growing end of the cleavage furrow. PMID- 11942609 TI - Interactions of Cdc4p, a myosin light chain, with IQ-domain containing proteins in Schizosaccharomyces pombe. AB - The fission yeast Schizosaccharomyces pombe undergoes cell division through a medially placed actomyosin-based contractile ring. One of the key components of this ring is the F-actin based motor protein myosin II. The myosin II heavy chain Myo2p has two light-chain-binding domains, IQl and IQ2, which bind the essential light chain, Cdc4p, and the regulatory light chain, Rlc1p. Previously, we have reported the characterization of cells expressing Myo2p lacking the IQ2 domain that facilitates Myo2p interaction with Rlc1p. In this study, we have created and characterized S. pombe strains carrying precise deletions of IQ1 and the entire neck region encompassing the IQ1 and IQ2 domains. Surprisingly, we found that the entire neck region of Myo2p is dispensable for Myo2p function. Cells deleted for IQ1, IQ2 and the entire neck region of Myo2p do not display any obvious cytoskeletal abnormalities. Immunofluorescence studies indicated that Cdc4p localizes at the ring in early and late mitotic cells in a strain in which interactions of Cdc4p with both the myosin II heavy chains (Myo2p and Myp2p) are abolished. Unlike mutations in Rlc1p that are suppressed by a simultaneous deletion of its binding site on Myo2p, mutations in the essential light chain Cdc4p are not suppressed by deletion of its binding sites on Myo2p, suggesting that Cdc4p may have additional partners essential for cytokinesis. Consistent with this, we provide evidence that two other IQ-domain containing actomyosin ring proteins, Rng2p (an IQGAP-related protein) and Myo51p (a type V myosin heavy chain), physically interact with Cdc4p. We concluded that Cdc4p, a novel myosin light chain, interacts with multiple actomyosin ring components to effect cytokinesis. PMID- 11942610 TI - The molecular mechanism of targeted vesicle transport in cytokinesis. AB - Recent studies have demonstrated that vesicle transport to cleavage furrow is indispensable for cytokinesis. Some animal and plant cells form distinct structures during cell division known as central spindle and phragmoplast, respectively. Several essential factors involved in the vesicle transport have been isolated so far. SNARE proteins and molecular motors play a central role in this process. For future research of cytokinesis, it is important to investigate these factors as well as cytoskeletal components of the contractile ring in detail. This review focuses on the molecular mechanism of targeted vesicle transport in cytokinesis. PMID- 11942611 TI - Identification of proteins involved in cytokinesis of Dictyostelium. AB - Dictyostelium is one of the model systems of choice for studying the cytokinesis of animal-type cells. Two types of cytokinesis mutants have been used to identify proteins involved in the cytokinesis of Dictyostelium: (1) type I, the mutant cells grow on substrates to produce giant multinucleate cells; (2) type II, the mutant cells divide nearly normally on substrates, but are unable to divide at all and get highly multinucleate in suspension culture. These two mutant types might correspond to the myosin II-independent and myosin II-including cytokinesis mechanisms, respectively. PMID- 11942612 TI - On the mechanism of cleavage furrow ingression in Dictyostelium. AB - The ability of Dictyostelium cells to divide without myosin II in a cell cycle coupled manner has opened two questions about the mechanism of cleavage furrow ingression. First, are there other possible functions for myosin II in this process except for generating contraction of the furrow by a sliding filament mechanism? Second, what could be an alternative mechanical basis for the furrowing? Using aberrant changes of the cell shape and anomalous localization of the actin-binding protein cortexillin I during asymmetric cytokinesis in myosin II-deficient cells as clues, it is proposed that myosin II filaments act as a mechanical lens in cytokinesis. The mechanical lens serves to focus the forces that induce the furrowing to the center of the midzone, a cortical region where cortexillins are enriched in dividing cells. Additionally, continual disassembly of a filamentous actin meshwork at the midzone is a prerequisite for normal ingression of the cleavage furrow and a successful cytokinesis. If this process is interrupted, as it occurs in cells that lack cortexillins, an overassembly of filamentous actin at the midzone obstructs the normal cleavage. Disassembly of the crosslinked actin network can generate entropic contractile forces in the cortex, and may be considered as an alternative mechanism for driving ingression of the cleavage furrow. Instead of invoking different types of cytokinesis that operate under attached and unattached conditions in Dictyostelium, it is anticipated that these cells use a universal multifaceted mechanism to divide, which is only moderately sensitive to elimination of its constituent mechanical processes. PMID- 11942613 TI - Genetic approaches to dissect the mechanisms of two distinct pathways of cell cycle-coupled cytokinesis in Dictyostelium. AB - Dictyostelium discoideum is a unique experimental organism which allows genetic analysis of the mechanism of cytokinesis of the animal type, and a number of mutations which affect cytokinesis in one way or other have been identified. Myosin II filaments accumulate in the equatorial region, and myosin II-null cells cannot divide in suspension, indicating that active, myosin II-dependent constriction of the cleavage furrow contributes to bisection of the cell. We refer to this method of cytokinesis as cytokinesis A. On substrates, however, myosin II-null cells divide efficiently in a cell cycle-coupled manner. This adhesion-dependent but myosin II-independent division method, which we termed cytokinesis B, is carried out by a pathway that is genetically distinct from that of cytokinesis A. Morphological analyses suggested that cytokinesis B is driven by radial traction forces generated along polar peripheries, which indirectly cause furrow ingression. Identification of two redundant pathways have allowed us to search genes involved in either pathway by mutagenizing cells which are already defective in one of the pathways. This approach enabled us to identify a number of novel cytokinesis-related genes, as well as to reclassify known genes as cytokinesis-related. PMID- 11942614 TI - Determination of division plane and organization of contractile ring in Tetrahymena. AB - In the molecular mechanism of division plane determination and contractile ring formation, Tetrahymena 85kDa protein (p85) is localized to the presumptive division plane before the formation of the contractile ring. p85 directly interacts with Tetrahymena calmodulin (CaM) in a Ca2+-dependent manner, and p85 and CaM colocalize in the division furrow. A Ca2+/CaM inhibitor N-(6-Aminohexyl) 5-chloro-1-naphthalenesulfonamide HCI (W7) inhibits the direct interaction between p85 and Ca2+/CaM. W7 also inhibits the localization of p85 and CaM to the division plane, and the formation of the contractile ring and division furrow. In addition, p85 binds to G-actin in a Ca2+/CaM dependent manner, but does not bind F-actin. Tetrahymena profilin is localized to division furrow and binds Tetrahymena elongation factor-1alpha (EF-1alpha). EF-1alpha, which induces bundling of Tetrahymena F-actin, is also localized to the division furrow during cytokinesis. The evidence also indicates that Ca2+/CaM inhibits the F-actin bundling activity of EF-1alpha, and that EF-1alpha and CaM colocalize in the division furrow. In this review, we propose that the Ca2+/CaM signal and its target protein p85 cooperatively regulate the determination of the division plane and the initiation of the contractile ring formation, and that profilin and a Ca2+/CaM-sensitive actin-bundling protein, EF-1alpha, play pivotal roles in regulating the organization of the contractile ring microfilaments. PMID- 11942615 TI - Cytokinesis in the C. elegans embryo: regulating contractile forces and a late role for the central spindle. AB - Genetic and molecular studies in the nematode Caenorhabditis elegans have identified multiple essential pathways that regulate and execute cytokinesis in early embryonic cells. These pathways influence both the microfilament cytoskeleton and the microtubule cytoskeleton. Microfilaments are enriched throughout the cell cortex at all times during the cell cycle in embryonic cells. Cortical microfilaments are required for multiple processes in embryonic cells, including polar body extrusion during meiosis, anterior-posterior axis specification by the sperm-donated microtubule-organizing center, and cytokinesis during mitosis. In addition to contractile apparatus proteins that are required positively for cleavage furrow ingression, the Nedd8 ubiquitin-like protein modification pathway negatively regulates contractile forces outside the cleavage furrow during cytokinesis. Another pathway that acts positively during cytokinesis involves the mitotic spindle. The central spindle, where anti parallel non-kinetochore microtubules overlap and are cross-linked, is required for a late step in cytokinesis, and other pathway(s) involved in membrane addition during cytokinesis may also require the central spindle. The amenability of C. elegans to classical genetics, the ease of reducing gene function with RNA interference, the completion of the genome sequence, and the availability of transgenic GFP fusion proteins that render the cytoskeleton fluorescent, all serve to make the early worm embryo an especially promising system for further advances in the identification of cytokinesis pathways, and in defining their interactions. PMID- 11942616 TI - Drosophila male meiosis as a model system for the study of cytokinesis in animal cells. AB - Drosophila male meiosis offers unique opportunities for mutational dissection of cytokinesis. This system allows easy and unambiguos identification of mutants defective in cytokinesis through the examination of spermatid morphology. Moreover, cytokinesis defects and protein immunostaining can be analyzed with exquisite cytological resolution because of the large size of meiotic spindles. In the past few years several mutations have been isolated that disrupt meiotic cytokinesis in Drosophila males. These mutations specify genes required for the assembly, proper constriction or disassembly of the contractile ring. Molecular characterization of these genes has identified essential components of the cytokinetic machinery, highlighting the role of the central spindle during cytokinesis. This structure appears to be both necessary and sufficient for signaling cytokinesis. In addition, many data indicate that the central spindle microtubules cooperatively interact with elements of the actomyosin contractile ring, so that impairment of either of these structures prevents the formation of the other. PMID- 11942617 TI - The pebble GTP exchange factor and the control of cytokinesis. AB - Several G proteins of the Rho family have been shown to be required for cytokinesis. The activity of these proteins is regulated by GTP exchange factors (GEFs), which stimulate GDP/GTP exchange, and by GTPase activating proteins (GAPs), which suppress activity by stimulating the intrinsic GTPase activity. The role of Rho family members during cytokinesis is likely to be determined by their spatial and temporal interactions with these factors. Here we focus on the role of the pebble (pbl) gene of Drosophila melanogaster, a RhoGEF that is required for cytokinesis. We summarise the evidence that the primary target of PBL is Rho1 and describe genetic approaches to elucidating the function of PBL and identifying other components of the PBL-activated Rho signalling pathway. PMID- 11942618 TI - Response of the cortex to the mitotic apparatus during polar body formation in the starfish oocyte of Asterina pectinifera. AB - In order to understand the mechanism of unequal division, polar body formation was investigated using the oocytes of the starfish, Asterina pectinifera. Cortical actin filaments were quantitatively measured after staining the maturing oocytes with fluorescently labeled phalloidin using a computer and image processing software. Before polar body formation, at first the actin filaments at the animal pole decreased and subsequently the animal pole bulged. On the other hand, actin filaments surrounding the animal pole increased gradually and made a cleavage furrow around the animal pole as the bulge grew. Then the furrow ingressed and finally a polar body formed. When the surface force was calculated according to the cell shape, the surface force decreased at the animal pole but the force at the contractile ring increased. When by micromanipulation the mitotic apparatus was detached and translocated to the cortex other than the animal pole, polar body formation occurred all over the cortex of the oocyte, which indicates that the response of the whole cortex to the mitotic apparatus is equal. These results indicate that the decrease in the actin filaments and surface force near the centrosome of the mitotic apparatus as well as the increase in the actin filaments and surface force at some distance of the centrosome is important for cytokinesis. PMID- 11942619 TI - The mechanism of cytokinesis: reconsideration and reconciliation. AB - The widely held models of cytokinesis contend that signals for cleavage are transmitted by astral microtubules, and that such signals elicit the assembly and contraction of an equatorial band of actin-myosin II filaments. However, experiments during the past decade have painted an increasingly complex picture, including strong evidence for the involvement of chromosomal passenger proteins and interzonal microtubules, and the involvement of not only cortical contraction but also cytoskeletal disintegration. The purpose of this article is to consider alternative models that might better accommodate both old and new observations. It is proposed that chromosomal passenger proteins undergo dynamic associations at centromeres during metaphase and are recruited from the cytoplasm to both astral and interzonal microtubules during anaphase. In addition, cytokinesis may be driven by global inward contractions coupled to a localized collapse of the equatorial cortex. PMID- 11942620 TI - Role of myosin light chain phosphorylation in the regulation of cytokinesis. AB - Phosphorylation of regulatory light chain (RMLC) of myosin II at Ser19/Thr18 is likely to play important roles in controlling the morphological changes seen during cell division of cultured mammalian cells. Phosphorylation of RMLC regulates the activity of myosin II, an essntial motor for cytokinesis, and phosphorylation of RMLC shows dramatic changes during mitosis. Two exzymes, myosin phosphatase and kinase, control phosphorvlation of RMLC. Myosin phosphatase is activated during mitosis, apparently as a result of mitosis specific phosphorylation of the myosin phosphatase targeting subunit (MYPT). This activation of myosin phosphatase is likely to result in RMLC dephosphorylation, causing the disassemly of stress fibers and focal adhesions during prophase. The phosphorylation of MYPT is lost in cyotokinesis, which would decrease myosin phosphatase activity. At the same time, ROCK (Rho-kinase) probably phosphorylates MYPT at its inhibitory sites, further decreasing the activity of myosin phosphatase. These changes in MYPT phosphorylation would raise RMLC phosphorylation, leading to the activation of myosin II for cyotokinesis. RMLC phosphorylation is also regulated by several RMLC kinases including ROCK (Rho kinase), MLCK and citron kinase, all of which are localized at cleavage furrows. Future studies should examine whether these multiple kinases are redundant or whether they control distinct aspects of cell division. PMID- 11942621 TI - Role of MgcRacGAP/Cyk4 as a regulator of the small GTPase Rho family in cytokinesis and cell differentiation. AB - To identify the key molecules that regulate differentiation of hematopoietic cells, we carried out retrovirus-mediated functional screening for cDNAs whose expression suppresses IL-6-induced differentiation of mouse myeloid leukemic M1 cells. From this screening, we obtained a full length cDNA encoding a mouse homologue of human MgcRacGAP. Overexpression of the anti-sense MgcRacGAP profoundly inhibited IL-6-induced macrophage-differentiation of M1 cells. On the other hand, overexpression of the full-length form of MgcRacGAP alone enhanced macrophage differentiation of M1 cells in response to IL-6, and induced macrophage differentiation of HL-60 leukemic cells. To determine how this protein regulates differentiation and proliferation, an antibody against MgcRacGAP was prepared. Immunohistochemical studies revealed that MgcRacGAP mainly localizes in the nucleus in interphase, accumulates on the mitotic spindle in metaphase, and is condensed in the midbody during cytokinesis. Overexpression of an N-terminal domain deletion mutant, which lacks the ability to localize to the midbody through association with tubulins, or a GAP-inactive mutant resulted in the formation of multinucleated cells in HeLa cells as well as in hemopoietic cells. Interestingly, MgcRacGAP in the midbody was phosphorylated probably on serine and threonine residues. These results indicate that MgcRacGAP regulates cytokinesis and cellular differentiation as a regulator of Rho family of GTPase and suggest that this process is controlled by some serine/threonine kinases. PMID- 11942622 TI - Role of chromosomal passenger complex in chromosome segregation and cytokinesis. AB - Chromosomal passenger proteins associate with chromosomes early in mitosis and transfer to the spindle at ana/telophase. Recent results show that aurora B/AIM-1 (aurora and Ipl1-like midbody-associated protein kinase), which is responsible for mitotic histone H3 phosphorylation, INCENP (Inner Centromere protein) and Survivin/BIR are in a macromolecular complex as novel chromosomal passenger proteins. Aurora B/AIM-1 can bind to Survivin and the C-terminal region of INCENP, respectively, and colocalizes with both proteins to the centromeres, midzone and midbody. Disruption of either aurora B/AIM-1 or INCENP function leads to sever defects in chromosome segregation and cytokinesis. Moreover, the formation of the central spindle through anaphase to cytokinesis is also disrupted severely. These data suggest that chromosomal passenger complex is required for proper chromosome segregation by phosphorylating histone H3, and cytokinesis by ensuring the correct assembly of the midzone and midbody microtubule. Chromosomal passenger protein complex may couple chromosome segregation with cytokinesis. PMID- 11942623 TI - Membrane lipid control of cytokinesis. AB - In the final stage of cell division, cytokinesis constricts and then seals the plasma membrane between the two daughter cells. The constriction is powered by a contractile ring of actin filaments, and scission involves rearrangement of the lipid bilayer of the cell membrane. We have shown that the lipid phosphatidylethanolamine (PE), which normally resides in the internal leaflet of the bilayer, is exposed on the external leaflet of the cleavage furrow as a result of enhanced transbilayer movement of the phospholipids during cytokinesis. To investigate the role of PE in cytokinesis, we employed two different approaches: manipulation of cell surface PE by a PE-binding peptide and establishment of a mutant cell line specifically defective in PE biosynthesis. Both approaches provide evidence that surface exposure of PE is essential for disassembly of the contractile ring at the final stage of cytokinesis. Based on these findings, we proposed that the transbilayer redistribution of PE plays a critical role in mediating coordinated movements between the contractile ring and the plasma membrane that are required for the proper progression of cytokinesis. PMID- 11942624 TI - Roles of septins in the mammalian cytokinesis machinery. AB - Septins comprise a eukaryotic guanine nucleotide binding protein subfamily which form filamentous heteropolymer complexes. Although mechanism of cytokinesis is diverged by species and tissues, loss of septin function results in the multinuclear phenotype in many organisms. Hence septin filaments beneath the cleavage furrow are hypothesized as a structural basis to ensure completion of cytokinesis. However, molecular mechanisms of septin assembly, disassembly and function have been elusive despite the potential importance of this ubiquitous cytoskeletal system. Meanwhile, growing evidence suggests that mammalian septins functionally or physically interact with diverse molecules such as actin, actin binding proteins, proteins of membrane fusion machinery, Cdc42 adapter proteins, a ubiquitin-protein ligase, and phosphoinositides. Careful integration of these data may provide insights into the mechanism of mammalian septin organization and functions in cytokinesis. PMID- 11942625 TI - Functions of unconventional myosins in the yeast Saccharomyces cerevisiae. AB - Unconventional myosins in the budding yeast play essential roles in diverse cellular functions, including endocytosis, actin organization, and polarized distribution of organelles. Several lines of evidence suggest that novel proteins, interacting with the unconventional myosins, regulate their functions. In this review, we focus on the functions of unconventional myosins from the point of view of myosin-interacting proteins. PMID- 11942626 TI - Diphosphorylated MRLC is required for organization of stress fibers in interphase cells and the contractile ring in dividing cells. AB - Activity of nonmuscle myosin II is regulated by phosphorylation of its regulatory light chain (MRLC). Phosphoryration of MRLC at both Thr18 and Ser19 (diphosphorylation) results in higher MgATPase activity and in promotion of the assembly of myosin II filaments than does that of MRLC at Ser19 (monophosphorylation) in vitro. To determine the roles of the diphosphorylated MRLC in vivo, we transfected three kinds of MRLC mutants, unphosphorylated, monophosphorylated and diphosphorylated forms (MRLC2(T18AS19A), substitution of both Ser19 and Thr18 by Ala; MRLC2(T18AS19D), Ser19 by Asp and Thr18 by Ala; and MRLC2(T18DS19D), both Ser19 and Thr18 by Asp, respectively), into HeLa cells. Cells overexpressing the mutant MRLC2(T18DS19D) contained a larger number of actin filament bundles than did those overexpressing the mutant MRLC2(T18AS19D). Moreover, cells overexpressing the nonphosphorylatable mutant MRLC2(T18AS19A) showed a decrease in the number of actin filament bundles. Taken together, our data suggest that diphosphorylation of MRLC plays an important role in regulating actin filament assembly and reorganization in nonmuscle cells. PMID- 11942627 TI - Expression of DNA methyltransferase (Dnmt1) in testicular germ cells during development of mouse embryo. AB - The DNA methylation pattern is reprogrammed in embryonic germ cells. In female germ cells, the short-form DNA methyltransferase Dnmt1, which is an alternative isoform specifically expressed in growing oocytes, plays a crucial role in maintaining imprinted genes. To evaluate the contribution of Dnmt1 to the DNA methylation in male germ cells, the expression profiles of Dnmt1 in embryonic gonocytes were investigated. We detected a significant expression of Dnmt1 in primordial germ cells in 12.5-14.5 day postcoitum (dpc) embryos. The expression of Dnmt1 was downregulated after 14.5 dpc after which almost no Dnmt1 was detected in gonocytes prepared from 18.5 dpc embryos. The short-form Dnmt1 also was not detected in the 16.5-18.5 dpc gonocytes. On the other hand, Dnmt1 was constantly detected in Sertoli cells at 12.5-18.5 dpc. The expression profiles of Dnmt1 were similar to that of proliferating cell nuclear antigen (PCNA), a marker for proliferating cells, suggesting that Dnmt1 was specifically expressed in the proliferating male germ cells. Inversely, genome-wide DNA methylation occurred after germ cell proliferation was arrested, when the Dnmt1 expression was downregulated. The present results indicate that not Dnmt1 but some other type of DNA methyltransferase contributes to the creation of DNA methylation patterns in male germ cells. PMID- 11942628 TI - Somatic lamins in germinal vesicles of goldfish (Carassius auratus) vitellogenic oocytes. AB - In fish and amphibians, B-type lamins are divided into somatic (B1, B2) and oocyte-type (B3) lamins. In this study, we purified nuclear lamins from rainbow trout erythrocytes, raised an anti-lamin monoclonal antibody (L-200) that recognizes goldfish somatic-lamins, and isolated cDNAs encoding goldfish B-type lamins (B1 and B2) from a goldfish cell culture cDNA library. Goldfish B-type lamins are structurally similar to lamins found in other vertebrates with minor amino acid substitutions in the conserved region. Western blot analysis showed that goldfish oocytes contained mainly GV-lamin B3 as well as some somatic lamins. Laser-confocal microscope observations revealed that lamin B3 was present only in GV nuclear lamina, whereas somatic lamins were present in dense fibrillar structures throughout nuclear gels of isolated GVs. Similar nuclear filamentous structures were also observed in GVs of paraffin embedded oocytes. Epitope mapping indicated that L-200 recognized a conserved region containing a short stretch of the alpha-helix coiled-coil rod domain (Y(E/Q)(Q/E)LL). A similar motif is also present in other cytoplasmic intermediate filaments (i.e., vimentin, desmin, peripherin and GFAP). Taken together, these findings suggest that lamins or lamin-related intermediate filaments are an important component of the interior architecture of goldfish vitellogenic oocyte nuclei (GVs). PMID- 11942629 TI - Molecular behavior in living mitotic cells of human centromere heterochromatin protein HPLalpha ectopically expressed as a fusion to red fluorescent protein. AB - We constructed stable mammalian cell lines in which human heterochromatin protein HP1alpha and kinetochore protein CENP-A were differentially expressed as fusions to red (RFP-HP1) and green fluorescent proteins (GFP-CENP-A). Heterochromatin localization of RFP-HP1 was clearly shown in mouse and Indian muntjac cells. By preparing mitotic chromosome spreads, the inner centromere localization of RFP HP1 was observed in human and Indian muntjac cells. To characterize its molecular behavior in living mitotic cells, time-lapse images of RFP-HP1 were obtained by computer-assisted image analyzing system, mainly with mouse cells. In G2 phase, a significant portion of RFP-HP1 diffused homogeneously in the nucleus and further dispersed into the cytoplasm soon after the nuclear membrane breakdown, while some remained in the centromeric region. Simultaneous observations with GFP-CENP A in human cells showed that RFP-HP1 was located just between the sister kinetochores and then aligned to the spindle midzone. With the onset of anaphase, once it was released from there, it moved to the centromeres of segregating chromosomes or returned to the spindle equator. As cytokinesis proceeded, HP1alpha was predominantly found in the newly formed daughter nuclei and again displayed a heterochromatin-like distribution. These results suggested that, although the majority of HP1alpha diffuses into the cytoplasm, some populations are retained in the centromeric region and involved in the association and segregation of sister kinetochores during mitosis. PMID- 11942631 TI - Transmission of through-the-lens Auger spectroscopy in an electron microscope. AB - In this paper, the transmission properties of through-the-lens Auger spectroscopy in an electron microscope using an electro-magnetic immersion lens are calculated. The effects of changes in acceptance angle and entrance slit of the spectrometer are discussed. It is shown that by including the effects of oscillations in final angle and radius, enhanced transmission is obtained under typical experimental conditions. PMID- 11942630 TI - AFM imaging in solution of protein-DNA complexes formed on DNA anchored to a gold surface. AB - A chemical procedure for anchoring DNA molecules to gold surfaces was used to facilitate the imaging of DNA and DNA-protein complexes in buffer solution by tapping mode atomic force microscopy (TMAFM). For preparing flat gold surfaces, a novel approach was employed by evaporating small amounts of gold onto freshly cleaved mica to give flat films that were stable under aqueous buffer conditions. The thickness of the investigated films ranged from 1 to 10 nm. For typical films of 4-6 nm, which were stable under aqueous buffer conditions, the root mean square (RMS) roughness ranged between 0.25 and 0.5 nm, as measured by atomic force microscopy (AFM). This roughness is comparable to that of obtained by the template stripped gold (TSG) technique, which is widely used in scanning probe microscopy but involves more preparation steps. In order to visualize DNA and DNA protein complexes by TMAFM, the DNA was chemisorbed to the gold surface through a linker carrying a terminal thiol group at the 5'-end of each of the DNA strands. The modified DNA fragments were bound to the gold films and imaged in buffer solution, while unmodified DNA could not be visualized. Since the DNA was not dried during the process, it can be assumed that its native conformation was retained. This mode of anchoring did not prevent interaction with proteins, as confirmed by the observation that the topology of a complex formed by adding the protein to a surface-anchored DNA was the same as that obtained by anchoring a pre-formed complex to the gold surface. We attribute this observation to the fact that the DNA is anchored to the gold surfaces only through its ends, therefore the DNA-support interaction is minimized but imaging is still possible. PMID- 11942632 TI - Real-space formulation of the quantum-mechanical electronic scattering under static n-fold axially symmetric electric and magnetic fields in a projection microscope. AB - A numerical technique is developed to compute electronic propagation in three dimensional potential-energy and vector-potential distributions, as required for the quantum-mechanical modelization of scattering in static electric and magnetic fields in a projection microscope. The technique is implemented in a transfer matrix and Green's functions general procedure to simulate field-emission and electronic projection microscopy. In particular, simulated observations of a transverse magnetic field confirms the occurrence of diffraction fringes that are oriented in the direction of the field. These diffraction fringes are mainly associated with the detection of a magnetic flux and tend to be more pronounced as this quantity increases. In the conditions of this paper, the smallest magnetic flux that was detected could be associated with a phase shift of around 2pi/8 rad. PMID- 11942633 TI - Phase retrieval in TEM using Fresnel images. AB - As an alternative to sideband holography in an electron microscope, methods for phase recovery by in-line holography utilising Fresnel images of aperiodic objects were tested with computed simulations and experimental data. Phases were recovered by minimising an error functional defined as a measure of the differences between experimental and calculated image intensities. The probability of convergence to local minima of the error function was reduced by increasing the ratio of known to unknown parameters, partly by use of several Fresnel images at different defoci, and also by an incremental relaxation of the phase bandwidth. Iterative methods did not converge reliably to a global minimum, but a conjugate gradient algorithm usually recovered the phases exactly, even for object arrays which included large phase variations. In practice, it was essential to use analytic expressions for the error gradients with respect to the phases, defoci and beam direction. Phase shifts of several radians were measured near the edge of a contaminated aperture. The factors that limit the accuracy and reliability of phase recovery from Fresnel images are discussed. PMID- 11942634 TI - A comparison between three-dimensional and two-dimensional grain boundary plane analysis. AB - An EBSD-based methodology for assessment of grain boundary planes, in particular sigma3 boundaries in face-centred cubic materials, has recently been devised. The method is based on trace analysis in a single, two-dimensional section rather than the more arduous three-dimensional method. The paper reports a data set of grain boundary planes in alpha-brass. mainly sigma3s, which have been analysed using both methods so that they can be compared and a recommendation made about the usefulness of the new method. It is shown that the new, two-dimensional method is a valuable tool in the analysis of grain boundary geometry, especially when used in conjunction with v/v(m), a parameter for assessing the proximity to the misorientation reference structure. PMID- 11942635 TI - Measurement of the volume expansion of a grain boundary by the phase method. AB - The volume expansion of a molybdenum twin boundary is measured by the phase method. The influence of experimental parameters such as specimen thickness and defocus is investigated. It is also shown that the presence of residual 3-fold astigmatism can induce important errors in the final value. PMID- 11942636 TI - Image reconstruction from electron and X-ray diffraction patterns using iterative algorithms: experiment and simulation. AB - The hybrid input-output iterative algorithm, which solves the phase problem for scattering from non-periodic objects, is reviewed for application to X-ray and electron diffraction data. Desirable convex constraints, including the sign of the scattering potential for electrons, and compact support, are discussed. The cases of complex and real exit-face wavefunctions, strong and weak phase objects, various supports, and the use of coherent focussed radiation are reviewed. Reconstruction of general complex objects requires accurate knowledge of the support, which should consist of two holes or a triangle in an opaque mask. The support boundaries should be as sharp as possible. Strong phase objects without absorption can be recovered if the support consists of one hole, is accurately known and has sufficiently sharp boundaries. Real and weak phase objects with absorption can be recovered without accurate knowledge of the support area if the support boundaries are sufficiently sharp and the support consists of one or more holes. A sign constraint on the scattering potential is used to recover weak phase objects. The experimental realization of theoretically desirable support conditions is discussed. A two-stage method of finding the support for complex objects is proposed. Experimental results from applying the Gerchberg-Saxton Fienup HiO-algorithm to coherent electron diffraction patterns are presented, using specially made e-beam lithographed support structures. Images with a resolution of about 5 nm are thus recovered from the intensities alone in coherent electron diffraction patterns from non-periodic objects. Limitations of the present experiments are identified and suggestions made for development of both X-ray and electron work. PMID- 11942637 TI - Electron nanodiffraction methods for measuring medium-range order. AB - Electron nanodiffraction in a scanning transmission electron microscopy (STEM) instrument with a beam diameter of the order of 1 nm can be used to assess the medium-range ordering, or the correlation of atom positions over distances of 1-3 nm, in thin films of disordered materials. Proposals are made for measurements of medium-range order by use of a thin annular detector in STEM, to give the equivalent of the variable-coherence microscopy of Treacy and Gibson (Acta Cryst. A 52 (1996) 212) and by measuring the correlation of diffraction intensities from neighboring nanometer-diameter regions. Two simpler methods for measuring the average dimensions of regions of correlated structure, by observing the persistence of diffraction spots as the beam is translated over the specimen, and by observing the dimensions of spots in greatly defocused diffraction patterns, have been proposed and applied to the study of thin films of amorphous carbon, silica and silicon nitride. PMID- 11942638 TI - Dual-color 4Pi-confocal microscopy with 3D-resolution in the 100 nm range. AB - We report the development of simultaneous two-color channel recording in 4Pi confocal microscopy. A marked increase of spatial resolution over confocal microscopy becomes manifested in 4Pi-confocal three-dimensional (3D) data stacks of dual-labeled objects. The fundamentally improved resolution is verified both with densely labeled fluorescence beads as well as with membrane labeled fixed Escherichia coli. The synergistic combination of dual-color 4Pi-confocal recording with image restoration results in dual-color imaging with a 3D resolution in the 100 nm range. PMID- 11942639 TI - The never-silent presses. PMID- 11942640 TI - A practical approach for STEM image simulation based on the FFT multislice method. AB - It has been demonstrated that a high-angle annular dark-field (HAADF) STEM technique gives an image resolving atomic columns. Due to the diffusion of this technique and an improvement of its resolution, a practical procedure for image simulation becomes important for a quantitative interpretation of the HAADF image. In this report a new practical scheme for a STEM image simulation is developed based on the FFT multislice algorithm. Here, a HAADF intensity due to thermal diffuse scattering (TDS) is calculated from the absorptive potential corresponding to high-angle TDS and the wave function equivalent to the propagating probe within the sample. Contrary to the commonly used Bloch wave method, a coherent bright-field intensity and a coherent HAADF intensity are also obtained straightforwardly. The HAADF image contrast calculated for GaAs is not simply proportional to Z2 as expected from the Rutherford scattering at high angle, and the As/Ga contrast ratio depends on the specimen thickness. This suggests that the generation of the HAADF signal is appreciably affected by the coherent dynamical scattering. The developed procedure here will have a definitive advantage over the Bloch wave approach for simulating the HAADF images expected from a defect and interface or amorphous materials, and also the HAADF image obtained by using a Cs-corrected microscope. This is because the former requires a huge super cell, while the latter needs a large objective aperture including a large number of incident beam directions. PMID- 11942641 TI - Reconstruction of the electric charge density in thin films from the contrast transfer function measurements. AB - The radial distribution of the beam-induced charge in thin films is investigated using the contrast transfer properties of the transmission electron microscope. The phase shift due to charging is measured as the phase difference between the contrast transfer functions of two photos taken with and without film at the back focal plane. Solving the inverse Laplace problem with this input data recovers the charge density of the measured film. The electric potential function in the whole area is reconstructed using the boundary integral method and the analytical solution of the Laplace equation for the electric potential is induced from unit step-wise surface charge. The phase shift of electron waves is derived in a weak lens approximation. In this way, the radial dependence of the charge density and the magnitude of the electrostatic potential at the thin film are obtained. The surface charge density reaches quasi-equilibrium state after the first 30 min of the electron beam pre-irradiation. The hydrocarbon contamination layer on the surface of the film is considered to be the main source of charging. An explanation of the qualitative behavior of the charge density, based on the contamination diffusion theory, is proposed. PMID- 11942642 TI - Ultra stable tuning fork sensor for low-temperature near-field spectroscopy. AB - We report on a distance control system for low-temperature scanning near-field optical microscopy, based on quartz tuning fork as shear force sensor. By means of a particular tuning fork-optical fiber configuration, the sensor is electrically dithered by an applied alternate voltage, without any supplementary driving piezo, as done so far. The sensitivity in the approach direction is 0.2nm, and quality factors up to 2850 have been reached. No electronic components are needed close to the sensor, allowing to employ it in a liquid He environment. The system is extremely compact and allows for several hours of stability at 5 K. PMID- 11942643 TI - Analysis of high resolution transmission electron microscope images of crystalline-amorphous interfaces. AB - For the analysis of images of homogeneous crystalline-amorphous interfaces we propose to average them along the interface obtaining the averaged interface image or the averaged intensity profile. Due to averaging, contrast components with the periodicity of the crystalline area of the image are extracted. Thus, the contrast features originating from the random overlap of the projected potentials of atoms in the amorphous layer are suppressed. It is shown that averaged images can be simulated by the multi-slice method using the novel approach to model the near interfacial amorphous structure by its mean atomic density distribution in front of the crystalline boundary. The crystalline structure is represented by its known atomic positions. We apply the proposed method to the investigation of the near interfacial short-range order in the c Si/ a-Ge crystalline-amorphous interface. PMID- 11942644 TI - Fluorescence anisotropy near-field scanning optical microscopy (FANSOM): a new technique for nanoscale microviscometry. AB - A near-field scanning optical microscope system was implemented and adapted for nanoscale steady-state fluorescence anisotropy measurement. The system as implemented can resolve approximately 0.1 cP microviscosity variations with a resolution of 250 nm laterally in the near field, or approximately 10 microm when employed in a vertical scanning mode. The system was initially used to investigate the extent of microviscous vicinal water over surfaces of varying hydrophilicity. Water above a cleaved mica surface was found to have a decreased microviscosity, while water above a hydrophobic surface showed no change (detection limit approximately 0.1 cP at approximately 30 + nm from the surface). PMID- 11942646 TI - Optimal experimental design of STEM measurement of atom column positions. AB - A quantitative measure is proposed to evaluate and optimize the design of a high resolution scanning transmission electron microscopy (STEM) experiment. The proposed measure is related to the measurement of atom column positions. Specifically, it is based on the statistical precision with which the positions of atom columns can be estimated. The optimal design, that is, the combination of tunable microscope parameters for which the precision is highest. is derived for different types of atom columns. The proposed measure is also used to find out if an annular detector is preferable to an axial one and if a C(s)-corrector pays off in quantitative STEM experiments. In addition, the optimal settings of the STEM are compared to the Scherzer conditions for incoherent imaging and their dependence on the type of object is investigated. PMID- 11942645 TI - Structure analysis of polymerized phospholipid bilayer by TED and direct methods. AB - This paper describes the use of elastic energy filtered transmission electron diffraction combined with Direct Methods in order to study the structure of thin Langmuir-Blodgett films of a radiation sensitive diacetylene polymer (DC8.9PC). We obtain a potential map for one projection by direct phasing of zone axis patterns, and discuss experimental problems and possible solutions. PMID- 11942647 TI - Determination of absolute configurations of crystal structures using electron diffraction patterns by means of least-squares refinement. AB - A simple method is reported to determine the absolute configuration of the crystal structure from electron diffraction patterns taken from very small areas. The method is based on the differences in the Friedel reflections, which are in general much larger than for X-rays due to the dynamical behaviour of the electron scattering. We express the absolute configuration with a parameter similar to the one Flack (Acta Cryst. A 39 (1983) 876) introduced in X-ray crystallography. This parameter is added to a refinement procedure that uses a multi-slice calculation to calculate diffraction patterns. The validity and strength of the method are shown with simulated and experimental data sets of GaN in the [0 1 0]-zone and a more complex compound, Ce5Cu19P12 in the [0 0 1] zone. PMID- 11942649 TI - Achievement motivation and academic performance among Turkish early and young adolescents in The Netherlands. AB - The authors conducted 3 studies in which Turkish and other adolescents in the Netherlands completed questionnaires that addressed the importance of collectivist cultural values for achievement motivation and educational outcomes. Compared with other minority group students and Dutch students, the Turkish students had stronger family-oriented achievement motivation. In addition, only among the Turks was ethnic identification positively related to family motivation. Furthermore, there were no differences in individual achievement motivation between the Turks and Dutch, and only among the former group was a combination of family motivation and individual motivation found. Also, for the Turks, family motivation was positively related to task-goal orientation, which mediated the relationship between family motivation and academic performance. Individual motivation was an independent predictor of performance. For the other ethnic minority groups and the Dutch, family motivation was not related to task goal orientation and performance. In addition, for these groups, task-goal orientation mediated the relationship between individual achievement motivation and performance. PMID- 11942648 TI - Third graders' understanding of core political concepts (law, nation-state, government) before and after teaching. AB - A curriculum of about 22 hr distributed over 3 months on some core political notions (law, the main Italian political offices, the state) was implemented in an Italian 3rd-grade class of 23 children. Another class of 20 children formed the comparison group. The workings of the executive system were exemplified by explaining the running of the school (which in Italy is state-run). Children's learning and retention of these notions were tested twice, 1 month and 10 months after the end of the curriculum. At pre-test, before implementation of the curriculum, the majority of the children did not have a concept of the political domain: They did not know of parliament and government, hardly distinguished between the law and other types of rules, and did not know that teachers, policemen, and judges are public servants. At post- and delayed post-test, most of the children in the experimental group had mastered the concepts, performing significantly better than the comparison group. Thus, the development of political understanding appears to be due to the acquisition of relevant domain specific knowledge rather than the emergence of across-the-board logic ability. PMID- 11942650 TI - Allocation of visual attention depends on type of precue. AB - To distinguish between theoretical concepts of how attention is allocated, participants were presented with different types of precues in 6 experiments. In 1 condition with 100% valid precues (Experiments 1 and 2), the time course of attention effects revealed that (a) higher accuracy was obtained with dynamic multiple-element precues (MEPs in which the unique element was defined by apparent motion) than with static MEPs, in which the elements did not move once they were presented (Cheal & Chastain, 1998); (b) a longer precue-target interval (stimulus-onset asynchrony; SOA) was needed to reach asymptote accuracy with dynamic MEPs than with dynamic single-element precues (SEPs); and (c) all dynamic precues (both MEPs and SEPs) resulted in a decline in accuracy at long SOAs. These results suggest that static and dynamic MEPs result in delayed engagement of attention relative to SEPs. Further, a decline in accuracy at long intervals is associated with static and dynamic SEPs and dynamic MEPs, but not with static MEPs. With irrelevant precues (Experiments 3 to 5), there was capture by precues in which the unique element moved briskly, smoothly, or abruptly, or simply flashed on and off, although there were differences in the amount of capture. The strongest capture occurred with smooth movement in static background elements and the weakest with smooth movement in abruptly moving background elements. It was shown in Experiment 6 that a static MEP will not capture attention if one element changes to a unique brightness near the time of precue onset, but if the element changes after 1,000 ms, it will capture attention. The authors suggest that different types of precues result in unequal influence of endogenous and exogenous components of attention, even when the same targets are used. In addition, they show that neither singleton detection mode nor contingent involuntary orienting is necessary for the capture of attention. PMID- 11942651 TI - MR metabolic imaging of brain lesions: lessons and caveats. PMID- 11942652 TI - Contrast medium concentration in epithelial mucosal cells after colonic instillation of lodixanol: a semiquantitative study indicating the route of permeation. AB - RATIONALE AND OBJECTIVES: Studies have shown that water-soluble radiographic contrast media may be useful as intestinal permeability probes. The current study was undertaken to provide information about the route by which such a permeability probe crosses the mucosal epithelium. MATERIALS AND METHODS: The authors administered 3 mL of iodixanol (iodine, 320 mg/mL) by means of an enema into the colons of (a) rats with experimental colitis induced 14 days earlier and (b) normal rats. At various intervals after the enema, laparotomy was performed and the colon was identified and longitudinally split immediately before cryofixation, without prior interruption of circulation. Freeze-dried cryosections were studied by means of electron microscopy and x-ray microanalysis. RESULTS: Approximately equal concentrations of iodine were found intracellularly and between the epithelial cells of the colonic mucosa. Ninety percent of analyzed cells had a normal sodium-potassium ratio and low levels of intracellular chlorine, indicating intact cell membranes. Nevertheless, many of these cells, in specimens from both normal rats and rats with colitis, contained high intracellular iodine concentrations. CONCLUSION: The transcellular route seems to be at least as important as the paracellular route for permeation of the highly water-soluble molecule iodixanol through the mucosal epithelium of both normal and inflamed rat colon. This finding may contradict previous findings in other water-soluble substances. PMID- 11942653 TI - Automated detection of polyps with CT colonography: evaluation of volumetric features for reduction of false-positive findings. AB - RATIONALE AND OBJECTIVES: To achieve high performance in computer-assisted diagnosis (CAD) of polyps with computed tomographic (CT) colonography, the authors (a) developed new gradient concentration and directional gradient concentration (DGC) features for differentiating between the true-positive and false-positive (FP) findings generated by the authors' CAD scheme, and (b) used receiver operating characteristic (ROC) analysis to quantify the differentiation performance of these and other volumetric features. MATERIALS AND METHODS: CT colonography was performed in 43 patients prone and supine with a helical CT scanner; there were 12 polyps in 11 patients. The polyp candidates generated by the authors' CAD scheme were characterized by nine statistics of six volumetric features, and the resulting 54 feature statistics were combined by a linear or quadratic discriminant classifier. The discrimination performance was measured with round-robin method by ROC analysis and the FP rate of the CAD scheme. RESULTS: The mean value of shape index (SI) yielded the highest individual ROC performance (area under the curve = 0.92). Among combinations, the mean values of SI and DGC and the variance of CT value yielded a high ROC performance (area under the curve = 0.95). With quadratic classifier, the sensitivity and FP rate of the case-based (data set-based) analysis was 100% (95%) with 2.4 FP findings per patient (1.7 FP findings per data set), respectively. CONCLUSION: Combination of the mean values of SI and DGC and the variance of CT value reduced the FP rate substantially without sacrificing sensitivity. These three features are potentially useful in improving the performance of the authors' CAD scheme for detecting polyps with CT colonography. PMID- 11942654 TI - Quantitative MR spectroscopic imaging of brain lesions in patients with AIDS: correlation with [11C-methyl]thymidine PET and thallium-201 SPECT. AB - RATIONALE AND OBJECTIVES: The purpose of this study was to examine the metabolic basis for magnetic resonance (MR) spectroscopic imaging findings in focal brain lesions of patients with acquired immunodeficiency syndrome (AIDS). MATERIALS AND METHODS: The authors combined MR spectroscopic imaging with [11C-methyl]thymidine positron emission tomography (PET) and thallium-201 single photon emission computed tomography (SPECT) in 10 patients. They sought a characteristic metabolite profile that would enable differentiation of benign lesions from primary central nervous system lymphoma and determined whether PET and SPECT results supported the MR spectroscopic imaging findings. RESULTS: Metabolite compositions of the lesions were similar, likely because the lesions were late stage and tended to be necrotic. Brain lesions in patients with AIDS were nonspecific regarding metabolic profile as determined with MR spectroscopic imaging. All patients with positive PET and SPECT studies had primary central nervous system lymphoma. CONCLUSION: PET and SPECT may be more reliable than MR spectroscopic imaging for differentiating benign lesions from primary central nervous system lymphoma in patients with AIDS. PMID- 11942655 TI - Outcome analysis of patients with acute pancreatitis by using an artificial neural network. AB - RATIONALE AND OBJECTIVES: The authors performed this study to evaluate the ability of an artificial neural network (ANN) that uses radiologic and laboratory data to predict the outcome in patients with acute pancreatitis. MATERIALS AND METHODS: An ANN was constructed with data from 92 patients with acute pancreatitis who underwent computed tomography (CT). Input nodes included clinical, laboratory, and CT data. The ANN was trained and tested by using a round-robin technique, and the performance of the ANN was compared with that of linear discriminant analysis and Ranson and Balthazar grading systems by using receiver operating characteristic analysis. The length of hospital stay was used as an outcome measure. RESULTS: Hospital stay ranged from 0 to 45 days, with a mean of 8.4 days. The hospital stay was shorter than the mean for 62 patients and longer than the mean for 30. The 23 input features were reduced by using stepwise linear discriminant analysis, and an ANN was developed with the six most statistically significant parameters (blood pressure, extent of inflammation, fluid aspiration, serum creatinine level, serum calcium level, and the presence of concurrent severe illness). With these features, the ANN successfully predicted whether the patient would exceed the mean length of stay (Az = 0.83 +/- 0.05). Although the Az performance of the ANN was statistically significantly better than that of the Ranson (Az = 0.68 +/- 0.06, P < .02) and Balthazar (Az = 0.62 +/- 0.06, P < .003) grades, it was not significantly better than that of linear discriminant analysis (Az = 0.82 +/- 0.05, P = .53). CONCLUSION: An ANN may be useful for predicting outcome in patients with acute pancreatitis. PMID- 11942656 TI - Optimal neural network architecture selection: improvement in computerized detection of microcalcifications. AB - RATIONALE AND OBJECTIVES: The authors evaluated the effect of optimal neural network architecture selection on the performance of a computer-aided diagnostic system designed to detect microcalcification clusters on digitized mammograms. MATERIALS AND METHODS: The authors developed a computer program to detect microcalcification clusters automatically on digitized mammograms. Previously, they found that a properly selected and trained convolution neural network (CNN) could reduce false-positive (FP) findings and therefore improve the accuracy of microcalcification detection. In the current study, they evaluated the effectiveness of the CNN optimized with an automated optimization technique in improving the accuracy of the microcalcification detection program, comparing it with the manually selected CNN. An independent test data set was used, which included 472 mammograms selected from the University of South Florida public database and contained 253 biopsy-proved malignant clusters. RESULTS: At an FP rate of 0.7 cluster per image, the film-based sensitivity was 84.6% for the optimized CNN, compared with 77.2% for the manually selected CNN. For clusters imaged on both craniocaudal and mediolateral oblique views, a cluster could be considered detected when it was detected on one or both views. For this case based approach, at an FP rate of 0.7 per image, the sensitivity was 93.3% for the optimized and 87.0% for the manually selected CNN. CONCLUSION: The classification of true and false signals is an important step in the microcalcification detection program. An optimized CNN can effectively reduce FP findings and improve the accuracy of the computer-aided detection system. PMID- 11942658 TI - Does medical school performance predict radiology resident performance? AB - RATIONALE AND OBJECTIVES: The authors performed this study to examine the relationship, if any, of a large number of measures of medical school performance with radiology residency performance. MATERIALS AND METHODS: Applications of 77 radiology residents enrolled from 1991 to 2000 were reviewed. Medical school grades, dean's letter summary statements, letters of recommendation, selection to Alpha Omega Alpha (AOA), and National Board of Medical Examiners (NBME) and U.S. Medical Licensing Examination (USMLE) Step 1 scores were recorded. Student t tests, analysis of variance, and correlation coefficients were used to examine the relationship between these measures of medical school performance and subsequent performance during radiology residency as determined by rotation evaluations, retrospective faculty recall scores, and American College of Radiology (ACR) and American Board of Radiology (ABR) examination scores. Resident performance was also correlated with prestige of the medical school attended. RESULTS: Preclinical grades of Honors or A; clinical grades of Honors or A in medicine, surgery, and pediatrics; and high NBME/USMLE scores strongly predicted success on the ABR written clinical examination but did not predict rotation performance. Most other measures of medical school performance, including outstanding Dean's letters and letters of recommendation, AOA selection during the senior year, and high medical school prestige did not predict high examination scores or superior rotation performance during residency. CONCLUSION: Success on the ABR examination can be predicted by medical school success in preclinical courses, some clinical courses, and USMLE examination scores. Dean's letters, letters of recommendation, AOA selection during the senior year, and medical school prestige do not appear to predict future resident performance as reliably. PMID- 11942657 TI - Normal adrenal gland: in vivo observations, and high-resolution in vitro chemical shift MR imaging-histologic correlation. AB - RATIONALE AND OBJECTIVES: The purpose of this study was to determine whether adrenal cortical lipid affects signal intensity on magnetic resonance (MR) images and to evaluate contrast between cortex and medulla. MATERIALS AND METHODS: From their clinical database, the authors selected 37 MR imaging studies of patients with adrenal adenomas. Two independent readers compared in-phase and fat suppressed T1-weighted images, looking for visible lipid-induced signal intensity loss in the adrenal gland. Six adrenal gland specimens obtained after radical nephrectomy were also studied with high-resolution MR imaging, including in phase, opposed-phase, and fat-suppressed T1-weighted images, and T2-weighted images. Adjacent histologic sections were stained with oil red O for neutral fats and with hematoxylin-eosin, and they were also viewed with polarization light microscopy. The relative amount of lipid was graded as mild, moderate, or intense, and the appearance of the cortex and medulla was compared with that on the MR images. RESULTS: On the 37 clinical MR studies, there was no visible signal intensity loss within the limbs of the ipsilateral adrenal glands. T2 weighted images of the adrenal specimens showed a thin high-intensity band, corresponding to the appearance of medulla on histologic slices. This could not be seen on any of the T1-weighted images. Region-of-interest measurements were nearly identical for in-phase and opposed-phase images. Histologic analysis showed abundant cortical lipid. CONCLUSION: Adrenal corticomedullary contrast can be depicted on high-resolution T2-weighted images but not on any T1-weighted images. There is abundant cortical lipid in adrenal specimens, but comparison of in-phase with opposed-phase MR images does not depict it. PMID- 11942659 TI - Feedback in radiologic education. PMID- 11942660 TI - Motivation of radiology residents: an interaction between personal and environmental factors. PMID- 11942661 TI - John L. Doppman: the clinician's radiologist. PMID- 11942662 TI - Image-guided percutaneous thermal ablation of bone tumors. PMID- 11942663 TI - Placement of a vena cava filter with an antecubital approach. AB - RATIONALE AND OBJECTIVES: The authors performed this study to evaluate the use of an antecubital venous approach for inferior vena cava (IVC) filter placement. MATERIALS AND METHODS: An IVC filter was placed in 26 patients (15 men, 11 women) in whom the antecubital vein was the preferred access site. An antecubital vein was accessed with ultrasound guidance and used for IVC filter placement. This same access site was used to place a peripherally inserted central catheter (PICC) in 17 of the 26 patients. Access was obtained via the basilic vein in 15 patients (58%), brachial vein in eight (31%), and cephalic vein in three (12%). RESULTS: The IVC filter was successfully placed in the infrarenal vena cava in all 26 patients (100%) by using an antecubital vein for access. All filters deployed appropriately without complication. No complications occurred during PICC placement. CONCLUSION: The IVC filter can be safely placed via an antecubital vein. When clinically necessary, this site can provide convenient access for the PICC placement. PMID- 11942664 TI - Future directions for the Academy of Radiology Research. PMID- 11942665 TI - Can I go to jail? A review of criminal liability for workplace injury or death. PMID- 11942666 TI - Fire hazard from filling portable gas cans in pickup trucks and cars. PMID- 11942667 TI - A Web-based resource for construction safety and health. PMID- 11942668 TI - Preventing musculoskeletal disorders in garment workers: preliminary results regarding ergonomics risk factors and proposed interventions among sewing machine operators in the San Francisco Bay Area. PMID- 11942669 TI - Postural sway measurements: a potential safety monitoring technique for workers wearing personal protective equipment. AB - This study investigated the use of postural sway measurements as a potential safety monitoring technique. Sixteen healthy male volunteers (age: 41.8+/-9.3 years) participated in this study. The level of neurophysiological strain and the ability to maintain upright balance for workers wearing personal protective equipment (PPE) were investigated. Three levels of PPE, the United States Environmental Protection Agency (USEPA)-defined levels A, B, and C, were worn in random order while performing two randomly ordered tasks, resting and a workload. After the workload task, the perceived exertion and discomfort were recorded. Postural sway, measured using a microprocessor-based force platform system, was assessed after each task. The variables of sway area and sway length were calculated from stabilograms. These measurements were taken for three sway tests: 1) Eyes open (EO), 2) Eyes closed on a four-inch foam (FC), and 3) a dynamic test with the subject reaching for a weight placed in front, lifting it to his or her chest, and lowering back to the shelf four times (RE). The heart rate was significantly higher for the workload task (p < 0.05), and the levels A and B PPE demonstrated significantly higher perceived exertion results than level C PPE (p < 0.05). The heart rate and perceived exertion were not significantly correlated with the sway measurements. Level A produced significant self-reported discomfort results for the upper back, lower back, and thighs. The sway variables showed significant differences with the PPE levels and the task. The workload task produced significantly higher sway length than the resting task (p < 0.05) for all test conditions. The PPE level B produced significantly higher sway length than PPE level A (p < 0.05) in the FC test condition. These results indicate that postural stability is altered with PPE use and with fatigued postural muscles. In summary, postural sway measurements may be used as a potential safety monitoring technique. PMID- 11942670 TI - Natural rubber latex allergy workers' compensation claims: Washington State healthcare workers, 1991-1999. AB - Occupational reaction to natural rubber latex experienced by healthcare employees was examined using data of all workers' compensation claims filed by state insured healthcare employees in Washington State for the period 1991-1999 (n = 65,703). As latex reaction is not a condition for which there are specific identification codes, these claims were estimated by coupling source and nature of injury records that were consistent with reactions to latex. It was found that the claim rate was on average 2.66 per 10,000 state-insured healthcare workers annually. The most common condition experienced was dermal (84.3%), and most common body part affected was the hand (70.0%). Because few claims cited respiratory or conjunctivitis as reaction experienced, little evidence was discovered to support that glove powder acted as a widespread latex allergen transmitter in healthcare environments. Most cases did not require indemnity payment for lost work time (81.2%), suggesting most reactions were minor. The average cost per natural rubber latex claim was $2,759.10, compared to $3,178.18 for the average healthcare worker claim for all causes. Overall, the average cost per state-insured healthcare worker employed during this time was under $0.74 per year. Nursing aides/orderlies were the most frequent healthcare occupation filing a claim (33.2%). The majority of claimants were female (87.9%), and unmarried workers (52.0%) filed slightly more claims than married workers (48.0%). In comparison with other workers' compensation claims filed by healthcare workers during this period, 0.34 percent of the total was potentially related to natural rubber latex, with other common healthcare workplace items cited more frequently as source of occupational injury. PMID- 11942671 TI - A synthetic vitreous fiber (SVF) occupational exposure database: implementing the SVF Health and Safety Partnership Program. AB - The Health and Safety Partnership Program is a voluntary workplace safety program for workers involved in the manufacture, fabrication, installation, and removal of glass wool and mineral wool products. This article describes one element of this Partnership Program, the development of an occupational exposure database that characterizes exposures by fiber type, industry sector, product type, and job description. Approximately 6000 exposure samples are included in the database, most of which were collected over the past decade, making it the most extensive and recent exposure data set on record for glass wool and mineral wool. The development of this database, as well as the initial results for exposure measurements segmented by product type and/or job description, are described. The current database shows that most applications and uses of glass wool and mineral wool involve exposures below the voluntary 1 f/cc permissible exposure limit, although some specific product types and job descriptions involve average exposures approaching the 1 f/cc limit. PMID- 11942672 TI - Occupational carbon monoxide poisoning in the State of Washington, 1994-1999. AB - Carbon monoxide poisonings continue to be significant and preventable for a number of work operations. This study assesses occupational carbon monoxide morbidity and mortality for the state of Washington based on a review of workers' compensation records for the years 1994-1999. The study characterizes sources, industries, and causative factors, and further attempts to identify work operations most at risk. Records were identified by both injury source and diagnostic codes. The study limits itself to non-fire-related carbon monoxide poisonings and primarily those from acute exposure. A decline in the number of claims was not evident, but the number of incidents per year showed a slight decline. Carbon monoxide poisonings were found to occur throughout all types of industries. The greatest number of claims was found in agriculture, followed by construction and wholesale trade, with these three accounting for more than half the claims and nearly half of the incidents. The more severe poisonings did not necessarily occur in industries with the greatest number of incidents. The major source for carbon monoxide poisoning was forklift trucks, followed by auto/truck/bus, portable saws, and more than 20 other sources. Fruit packing and storage had the highest number of incidents mostly due to fuel-powered forklift activity, with nearly half of the incidents occurring in cold rooms. Adverse health effects as measured by carboxyhemoglobin, hyperbaric oxygen treatment, unconsciousness, and number and cost of claims were indexed by source. Though several specific work operations were identified, the episodic nature of carbon monoxide poisonings, as well as the diverse industries and sources, and the opportunity for a severe poisoning in any number of operations, poses challenges for effective intervention. PMID- 11942673 TI - Evaluation of occupational exposure to toxic metals released in the process of aluminum welding. AB - The objective of this study was to evaluate occupational exposure to welding fumes and its elements on aluminum welders in Polish industry. The study included 52 MIG/Al fume samples and 18 TIG/Al samples in 3 plants. Air samples were collected in the breathing zone of welders (total and respirable dust). Dust concentration was determined gravimetrically, and the elements in the collected dust were determined by AAS. Mean time-weighted average (TWA) concentrations of the welding dusts/fumes and their components in the breathing zone obtained for different welding processes were, in mg/m3: MIG/Al fumes mean 6.0 (0.8-17.8), Al 2.1 (0.1-7.7), Mg 0.2 (< 0.1-0.9), Mn 0.014 (0.002-0.049), Cu 0.011 (0.002 0.092), Zn 0.016 (0.002-0.14), Pb 0.009 (0.005-0.025), Cr 0.003 (0.002-0.007), and TIG/Al fumes 0.7 (0.3-1.4), Al 0.17 (0.07-0.50). A correlation has been found between the concentration of the main components and the fume/dust concentrations in MIG/Al and TIG/Al fumes. Mean percentages of the individual components in MIG/Al fumes/dusts were Al: 30 (9-56) percent; Mg: 3 (1-5.6) percent; Mn: 0.2 (0.1-0.3) percent; Cu: 0.2 (< 0.1-1.8) percent; Zn: 0.2 (< 0.1-0.8) percent; Pb: 0.2 (< 0.1-1) percent; Cr: < 0.1 percent. The proportion of the respirable fraction in the fumes and their constituents varied between 10 percent and 100 percent. The results showed that MIG/Al fumes concentration was 1.2 times higher than the American Conference of Governmental Industrial Hygienists (ACGIH) threshold limit value (TLV), and the index of the combined exposure to the determined agents was 2.3 (0.4-8.0), mostly because of high Al2O3 contribution. The background concentrations of the components (ca. 5-10 times lower than those in the breathing zone of the welders) did not exceed the Polish MAC value. The elemental composition of total and respirable fume/dust may differ considerably depending on welding methods, the nature of welding-related operations, and work environment conditions. PMID- 11942674 TI - Validation of a job-exposure matrix for assessment of utility worker exposure to magnetic fields. AB - The aim of this study was to evaluate a 50-Hz electromagnetic field job-exposure matrix used in epidemiological studies of a nationwide cohort of utility workers in Denmark. We compared a job-exposure matrix that distinguished four categories of exposure to 50-Hz time-weighted average (TWA) magnetic fields: low (< 0.1 microT), medium (0.1-0.29 microT), high (0.3-0.99 microT) and very high (> 1.0 microT) of utility company employees with 196 measurements of 8-h exposure for 129 workers in this industry. The 129 workers were selected from the following five main work environments: generation facilities, transmission lines, distribution lines, substations, and other electrically and non-electrically relates jobs. This study shows that the job-exposure matrix can be expected to introduce misclassification mainly between adjacent categories of exposure. Thus, the distribution of measurements of exposure to 50-Hz magnetic fields was similar for workers in the medium and the high exposure matrix categories. But the two extreme categories satisfactorily separate low and very highly exposed workers. The study shows that epidemiological use of this job-exposure matrix might combine the two intermediate categories of exposure. If the sample size in extreme categories provides enough power, a study in which this job-exposure matrix is used should allow detection of a true association between exposure to 50-Hz magnetic field and disease. PMID- 11942675 TI - Lignans and neolignans from Myristica argentea Warb. AB - The petrol extract from mace of Myristica argentea Warb. afforded six phenylpropenes, three lignans, three neolignans and a dilignan, bis erythro argenteane (4) or rel-(8R,8'S,8"S,8"'R)-5',5"'-bis(7-(3,4-methylenedioxyphenyl) 7'-(4'-hydroxy-3'-methoxyphenyl)-8.8'-lignane]. The last-named compound is a new natural product. PMID- 11942677 TI - A new drimane from the heartwood of the Japanese yew, Taxus cuspidata. AB - A new drimane sesquiterpenoid was isolated from the heartwood of the Japanese yew, Taxus cuspidata Sieb. et Zucc. Its structure was established as 1beta acetoxyisodrimeninol (1beta-acetoxy-11,12-epoxy-7-drimen-11-ol, 1). This compound was gradually oxidized to give y-lactone (1beta-acetoxy-8-drimeno-11,12-lactone, 4). PMID- 11942676 TI - New cytotoxic cucurbitacins from the pericarps of Trichosanthes tricuspidata fruits. AB - An extract of the fruits of Trichosanthes tricuspidata collected in North Vietnam was found cytotoxic in KB cells. A bioassay-guided fractionation led to the isolation of a series of cucurbitacins of which two are new: tricuspidatin and 2 O-glucocucurbitacin J. Their isolation and structure determination are described. PMID- 11942678 TI - An in vitro screening method for DNA cytosine-C5-methylase inhibitor. AB - A specific inhibitor of DNA cytosine C5-methylases would be useful for studying genomic imprinting, X-chromosome inactivation, carcinogenesis, and regulation of tissue-specific gene expression, for these physiological phenomena appears to be regulated through DNA methylation in promoter sequences. This paper reports a novel convenient in vitro assay method for screening DNA cytosine C5-methylase inhibitor. Our method uses a commercially available Hae III methylase (cytosine C5 methylase), its corresponding Hae III endonuclease, and lambda DNA as their substrate. PMID- 11942679 TI - A novel flavonol lyxoside of Orostachys japonicus herb. AB - A novel flavonol lyxoside was isolated from the aerial part of Orostachys japonicus (Crassulaceae). Its structure was determined as gossypetin 8-O-alpha-D lyxopyranoside using several spectral data and chemical methods. Lyxoside of flavonoid was isolated for the first time from nature. PMID- 11942680 TI - Diterpenes from Sideritis trojana. AB - Six known ent-kaurene, a new ent-kaurane and a new pimarane diterpenes were isolated from Sideritis trojana. The structures of new compounds were determined as ent-7alpha-15beta,16beta-epoxykaurane (1), and ent-2alpha-hydroxy-8(14),15 pimaradiene (2) along with the known compounds siderol (3), sideridiol (4), 7 epicandicandiol (5), isocandol B (6), candol A acetate (7) ent-7alpha-acetoxykaur 15-ene (8) by IR, 1D and 2D NMR techniques and HRMS. PMID- 11942681 TI - Licamichauxiioic-A and -B acids--two ent-kaurene diterpenoids from Licania michauxii. AB - Bioassay-guided fractionation allowed the isolation of two new cytotoxic ent kaurene diterpenoids, licamichauxiioic-A and -B acids (1 and 2) from the root extract of Licania michauxii Prance (Chrysobalanaceae). They were characterized as ent-15-oxo-9(11),16-kauradien-19-oic acid (1) and ent-15-oxo-13(14),16 kauradien-19-oic acid (2) by various spectroscopic methods, in particular, 1D and 2D NMR spectra, and chemical evidence. PMID- 11942682 TI - Diterpenoids from Salvia ceratophylla. AB - Salvia ceratophylla L. has yielded four known and two new diterpenoids together with two triterpenic acids, a steroid and a flavone. The structures of the compounds were established by spectroscopic analyses. One of the known compounds candidissiol exhibited a high antibacterial activity against Staphylococcus epidermidis and Proteus mirabilis. PMID- 11942683 TI - A new approach for asymmetric synthesis of (R)-3-methylpyrrolidine alkaloids from (S)-malic acid. AB - Diastereoselective methylation of dimethyl (S)-malate 7, followed by two, three step reductive de-hydroxylation procedures afforded dimethyl (R)-2 methylsuccinate 11 in 80.2% e.e. and 84.7% e.e., respectively. The latter compound was further transformed into the natural enantiomers of the ant venom alkaloids (R)-leptothoracine 1 and (R)-3-methyl-N-(2-phenylethyl)-pyrrolidine 2. PMID- 11942684 TI - Chemotaxonomic features associated with flavonoids of cannabinoid-free cannabis (Cannabis sativa subsp. sativa L.) in relation to hops (Humulus lupulus L.). AB - The major flavonoids present in the leaves and flowers of the cannabinoid-free cannabis (Cannabis sativa subsp. sativa L.) cultivars Felina and Futura are orientin (1), vitexin (2), luteolin-7-O-beta-D-glucuronide (3), and apigenin-7-O beta-D-glucuronide (4), while prenylated flavonoids, to which the potent estrogenicity of hops (Humilus lupulus L.) is associated, are absent. The different composition of flavonoids has chemotaxonomic value. PMID- 11942685 TI - 1,5-dihydroxyanthraquinones and an anthrone from roots of Rumex crispus. AB - From the roots of Rumex crispus, two known anthraquinones and a new one together with a new anthrone were isolated and the structures of compounds 1-4 were elucidated by spectroscopic means. The singlet oxygen generation capacity was tested with 1,3-diphenylisobenzofuran (DPBF) for compounds 1-4. PMID- 11942686 TI - Extraction studies of Tabernanthe iboga and Voacanga africana. AB - The root bark of Tabernanthe iboga contains ibogaine as its predominant alkaloid and has been an important source of it. Ibogaine is used experimentally to interrupt drug addiction and allow therapeutic intervention, but is currently unaffordable to doctors in less economically developed countries. To meet this need, an extraction of alkaloids from T. iboga root bark was optimized and simplified to use only diluted vinegar and ammonia, and was successfully applied to related alkaloids from Voacanga africana bark also. The alkaloids were converted to their hydrochlorides and purified, and the minor alkaloids were recovered. PMID- 11942687 TI - Bioconversion of ilicic alcohol and derivatives with cultures of filamentous fungi. AB - Aspergillus niger cultures monohydroxylate ilicic alcohol in C-3 in a cis position with respect to the methyl groups at C-4 and C-10, and trans position with respect to the hydroxyl group of C-4. Furthermore, Aspergillus niger cultures reduce ilicic aldehyde to its corresponding alcohol. PMID- 11942688 TI - Specific GABA binding in the adrenals and blood corticosteroid levels in stress in intact rats and rats with changes in the functional activity of the hypothalamo-pituitary-adrenal system. AB - Thirty minutes of swimming led to decreases in the specific binding of GABA to adrenal plasma membranes in intact rats, but not in rats repeatedly given hydrocortisone to inhibit the baseline activity of the hypothalamic-pituitary adrenal cortex system, where the level of reception of this amino acid was significantly greater than in controls. Pre-stress administration of baclofen and gamma-L-aminobutyryltaurine had no effect on specific GABA binding, even though these agents affected blood corticosterone concentrations. PMID- 11942689 TI - Model studies of the mechanisms of tuning of visual cortex neurons to incomplete cross-shaped figures. AB - Numerical simulation modeling of the receptive fields of visual cortex neurons able to detect cross-shaped figures with masked central or peripheral areas was performed. Receptive field models of two types were considered: those with antagonistic and cooperative interactions between the center and the periphery. Model neurons with receptive fields with reciprocal (antagonistic) interactions produced greater responses to peripheral or central crosses than to complete crosses. Studies using the model showed that the basis of this type of tuning could be provided by a disinhibition mechanism: blockade of the inhibitory zones in the center or periphery of the receptive field by activation of a lateral disinhibitory zone. A model with cooperative interactions between the center and periphery of the receptive field was also studied, in which responses to complete crosses were summed from the responses to the peripheral and central parts. Tuning of these model receptive fields was comparable to the sensitivity of real visual cortex neurons to the shape, size, and orientation of figures. The properties of model receptive fields (configuration, localization, and weightings of the various zones) allowing simulation of the properties of cat visual cortex field 17 neurons sensitive to the orientation and configuration of incomplete cross-shaped figures were identified. PMID- 11942690 TI - Responses of hippocampal neurons at different stages of acquisition of conditioned reflex avoidance in rats. AB - The electrophysiological properties of hippocampal slices from trained and untrained rats presented with identical numbers of combinations during acquisition of a conditioned two-sided avoidance reaction were compared. Associative learning (presentation of combinations of conditioned and unconditioned stimuli) was accompanied by gradual increases in the amplitude of the population spike in hippocampal field CA1. However, achievement of the learning criterion was accompanied by a decrease in the amplitude of the population spike. The same type of difference between groups was seen in field CA1 using low-intensity stimulation of Schaffer collaterals, while there was no change in the amplitude at maximum-intensity stimulation. After the rats achieved the learning criterion, the level of facilitation in conditions of paired stimuli was significantly greater than in untrained animals and also as compared with the passive and active controls. These data provide evidence supporting the hypothesis that changes in the efficiency of synaptic transmission occur during the learning process. However, the direction of changes depends on the extent of training. PMID- 11942691 TI - Selective effects of an NMDA glutamate receptor antagonist on the sensory input from chemoreceptors in the snail's head during acquisition of nociceptive sensitization. AB - Experiments on semi-intact preparations from common snails were used to study the characteristics of the actions of MK-801, an antagonist of NMDA glutamate receptors, on the plasticity of various sensory inputs to defensive behavior command neurons LP11 and RP11 during acquisition of nociceptive sensitization. Application of sensitizing stimuli to the head or foot of control snails led to depression of neuron responses to tactile and chemical sensory stimulation during the short-term stage and marked facilitation of these responses during the long term stage of sensitization. Application of sensitizing stimuli to the snail's head during administration of MK-801 led to marked depression of responses to chemical stimulation of the head in both the short-term and long-term stages of sensitization. In addition, blockade of NMDA receptors during application of sensitizing stimuli to the foot or head had no effect on changes in neuron responses elicited by chemical stimulation of the snail's foot and by tactile stimulation of the foot or head. It is suggested that NMDA-like glutamate receptors are selectively involved in the mechanism of induction of plasticity of synaptic inputs to command neurons LP11 and RP11, excited by chemical sensory stimulation of the head--a skin receptor zone specific for these neurons. PMID- 11942692 TI - Cellular gliosis of the white matter of the human brain and its importance in the pathogenesis of focal epilepsy. PMID- 11942693 TI - Steroids in the control of reproductive function in fish. AB - Serum and coelomic fluid sex steroid hormone levels were measured in the sturgeon (Acipenseridae) at the onset of anadromous migration and maturation. Cortisol and testosterone levels in coelomic fluid were lower than in serum; conversely, progesterone levels were higher in coelomic fluid than in blood. Specific androgen and estrogen binding in the cytosol of different parts of the brain and in the gonads of fish were different in the pre-spawning state and after ovulation. Before spawning, the highest levels of specific androgen binding were in the forebrain, where levels decreased significantly after ovulation. Specific estrogen binding in the hypothalamus was significantly higher after maturation and ovulation. Correlations were established between sex steroid concentrations in the blood and levels of specific binding in the brain and gonads. PMID- 11942694 TI - Mechanisms of the non-neurotransmitter actions of acetylcholine in the neuromuscular apparatus. AB - Experiments on isolated phrenic-diaphragmatic preparations from rats showed that low acetylcholine concentrations increase the work capacity of exhausted muscle by increasing the level of evoked quantum transmitter release; acetylcholine also induced hyperpolarization of muscle fiber membranes. The effects of acetylcholine persisted for long periods of time. The modulating actions of acetylcholine are mediated by structures with different pharmacological characteristics from those of typical n- and m-cholinoreceptors; these mechanisms involved ouabain-sensitive isoforms of Na+,K+-ATPase and, perhaps, membrane K+ channels. The data obtained here support the possible existence of long-term neuronal regulation of the efficiency of neuromuscular transmission involving non-quantal acetylcholine, these mechanisms presumably developing differently in muscle fibers with different functional characteristics and abilities to adapt to physiological loading. PMID- 11942695 TI - Blocker studies of the functional architecture of the NMDA receptor channel. AB - Blockade of ion channels passing through the NMDA receptors of isolated rat hippocampus pyramidal neurons with tetraalkylammonium compounds, 9-aminoacridine, and Mg2+ was studied using patch-clamp methods in the whole-cell configuration. Currents through NMDA channels were evoked by application of 100 microM aspartate in magnesium-free medium containing glycine (3 microM) to neurons. Analysis of the kinetics, charge transfer, and relationships between the extent of suppression of stationary currents on the one hand and membrane potential, agonist concentration, and blocker concentration on the other showed that blockers had different effects on the closing, desensitization, and agonist dissociation of NMDA channels. The size of the blocker was found to be the decisive factor determining its action on the gating functions of NMDA channels: larger blockers prevented closure and/or desensitization of the channel; smaller blockers only had partial effects on these processes, while the smallest blockers had no effect at all. These experiments showed that the apparent affinity of the blocker for the channel (1/IC50) depended not only on the microscopic equilibrium dissociation constant (Kd), but also on the number of blocker binding sites, their mutual influences, and, of particular importance, the interaction of the blocker with the gating structures of the channel. These data led us to propose hypotheses relating to the geometry of the NMDA channel and the structure of its gating mechanism. The channel diameter at the level of activated gates was estimated to be 11 A. PMID- 11942696 TI - Structural characteristics of ionotropic glutamate receptors as identified by channel blockade. AB - The channels of four types of ionotropic glutamate receptor (NMDA receptors and Ca-permeable AMPA receptors of rat brain neurons, and cation-selective receptors from mollusk neurons and insect postsynaptic muscle membranes) and two subtypes of nicotinic cholinoreceptor (from frog neuromuscular junctions and cat sympathetic ganglia) were studied. The structural characteristics of channels determining their susceptibility to blockade by organic mono- and dications were identified. These studies used homologous series of adamantane and phenylcyclohexyl derivatives. These experiments showed that the receptors studied here could be divided into two groups. The first group included the AMPA receptor and the mollusk and insect receptors. These were characterized by the lack of effect on the part of monocations and a strong relationship between the activity of dications and the distance between nitrogen atoms. The second group included the NMDA receptor and both subtypes of the nicotinic cholinoreceptor (muscular and neuronal). Here, conversely, the activity of monocations and dications, regardless of their lengths, were essentially identical. A model for the binding sites of blockers in channels is proposed, which takes these observations into account. PMID- 11942697 TI - Dopaminergic transmission in the rat striatum in vivo in conditions of pharmacological modulation. AB - The effects of pharmacological modulation of striatal dopaminergic neurotransmission were studied in freely mobile rats by intracerebral microdialysis and HPLC to assay dopamine and dopamine metabolite levels and the rate of dopamine synthesis, in combination with observations of stereotypical behavior. Inhibition of catechol O-methyltransferase (COMT) with tolcapone led to increases in extracellular dopamine levels only when the baseline dopamine level was elevated by administration of L-3,4-dihydroxyphenylalanine in combination with the decarboxylation inhibitor carbidopa. Increases in dopamine levels in striatal dialysates by blockade of reuptake were enhanced by inhibition of metabolic degradation of dopamine by tolcapone, a selective catechol O methyltransferase inhibitor. GBR-12909, a blocker of the dopamine transporter, increased extracellular dopamine and induced motor stereotypy. Both of these effects were potentiated by tolcapone. The rate of dopamine biosynthesis decreased when reuptake was inhibited. These data provide evidence for the key role of the dopamine transporter in maintaining neurochemical homeostasis at the synaptic level. PMID- 11942698 TI - The effects of immunization against cholecystokinin fragment 30-33 in the behavior of white rats. AB - Active immunization of white rats with cholecystokinin-4 covalently linked to the antigen carrier BSA evoked long-lasting changes in the rats' behavior, which were in the opposite direction to the anxiogenic effects of cholecystokinin-4 itself, showing that immunization had anxiolytic effects. Immunoenzyme analysis demonstrated the presence of antibodies to cholecystokinin-4 in the serum of immunized rats. These data are interesting from the point of view of correcting pathological anxiety and fear states by inverse immunoregulation. PMID- 11942700 TI - Energy recovery from grass using two-phase anaerobic digestion. AB - Municipal solid wastes are major sources of air, water and soil contamination. There is a need for alternative waste management techniques to better utilize the waste and minimize its adverse environmental impact. A two-phase pilot-scale bio fermentation system was used to evaluate the feasibility of producing methane from grass waste, a major constituent of solid wastes. The bi-phasic system consists of a solid phase and a methane phase. Leachate is re-circulated through the solid phase until a desired level of volatile fatty acid (VFA) is accumulated in the leachate. The leachate is then transferred to the methane reactor where the VFA is converted to methane. The results showed that 67% of the volatile solids in the waste can be converted into soluble chemical oxygen demand in a period of six months. The system produced an average of 0.15 m3 of methane per kg of grass. The average methane concentration in the produced gas was 71%. A mathematical model was developed to estimate the methane and carbon dioxide concentrations in the gas phase as a function of reactor properties. PMID- 11942699 TI - Serotonin modulates oscillations of the membrane potential in isolated spinal neurons from lampreys. AB - Studies were performed on spinal neurons from lampreys isolated by an enzymatic/mechanical method using pronase. The effects of 100 microM serotonin (5 HT) on membrane potential oscillations induced by a variety of excitatory amino acids were studied. 5-HT was found to depolarize branched cells (presumptive motoneurons and interneurons) by 2-6 mV without inducing membrane potential oscillations. However, when oscillations were already present because of an excitatory amino acid, 5-HT changed the parameters of these oscillations, increasing the amplitudes of all types of oscillations, increasing the frequency of irregular oscillations, and increasing the duration of the depolarization plateaus accompanied by action potentials. Serotonin modulation of the effects of excitatory amino acids and the electrical activity of cells in the neural locomotor network facilitates motor activity and leads to increases in the contraction of truncal muscles and more intense movements by the animal. The possible mechanisms of receptor coactivation are discussed, along with increases in action potential frequency and changes in the parameters of the locomotor rhythm. PMID- 11942701 TI - Gas production and transport in artificial sludge depots. AB - This paper presents a study to determine the impact of gas production in dredging sludge on the storage capacity of artificial sludge depots. Gas is produced as a result of the decomposition of organic material present in dredging spoil. This process, in which methane and carbon dioxide are formed, may lead to expansion of sludge layers, partly or even completely counterbalancing consolidation. The study shows that, even with a very conservative estimation of the rate of gas production, accumulation of gas occurs as convective and diffusive transport proceed very slowly. Nucleation of gas bubbles occurs already at a limited oversaturation of pore water. During their growth, bubbles push aside the surrounding grain matrix. Resulting stresses may initiate cracks around bubbles. If these cracks join, they may form channels stretching out to the depot surface and along which gas may escape. However, channels are only stable to a limited depth below which bubble accumulation may continue. The gas content at which sufficient cracks and channels are formed to balance the rate of gas production with the rate of outflow strongly depends on the constitutive properties of the dredging sludge considered. In sludge with a high shear strength (> 10 kPa), stable channels are created already at low deformations. However, a large expansion may occur in sludge with a low strength. The present study shows that accumulation of gas may continue until a bulk density less than that of water is attained. This is equivalent to a gas fraction of about 25-37%, depending on the initial water content of the sludge. Only then can gas escape as a result of instabilities in the sediment matrix. This should be well taken into account during the design and management of artificial depots. PMID- 11942702 TI - A comparison of refuse attenuation in laboratory and field scale lysimeters. AB - For this study, small and middle scale laboratory lysimeters, and a large scale field lysimeter in situ in Shanghai Refuse Landfill, with refuse weights of 187,600 and 10,800,000 kg, respectively, were created. These lysimeters are compared in terms of leachate quality (pH, concentrations of COD, BOD and NH3-N), refuse composition (biodegradable matter and volatile solid) and surface settlement for a monitoring period of 0-300 days. The objectives of this study were to explore both the similarities and disparities between laboratory and field scale lysimeters, and to compare degradation behaviors of refuse at the intensive reaction phase in the different scale lysimeters. Quantitative relationships of leachate quality and refuse composition with placement time show that degradation behaviors of refuse seem to depend heavily on the scales of the lysimeters and the parameters of concern, especially in the starting period of 0 6 months. However, some similarities exist between laboratory and field lysimeters after 4-6 months of placement because COD and BOD concentrations in leachate in the field lysimeter decrease regularly in a parallel pattern with those in the laboratory lysimeters. NH3-N, volatile solid (VS) and biodegradable matter (BDM) also gradually decrease in parallel in this intensive reaction phase for all scale lysimeters as refuse ages. Though the concrete data are different among the different scale lysimeters, it may be considered that laboratory lysimeters with sufficient scale are basically applicable for a rough simulation of a real landfill, especially for illustrating the degradation pattern and mechanism. Settlement of refuse surface is roughly proportional to the initial refuse height. PMID- 11942703 TI - Leaching properties of stabilised/solidified cement-admixtures-sewage sludges systems. AB - One of the main objectives of this work is to present an effective alternative for the final destination of sludge from urban waste water treatment plants by its use as a component of mortar or concrete. A binding and stabilizing matrix of sludge-cement and sludge-cement-coal fly-ash was investigated and the effects of various percentages of waste and binder, on the behavior of sludge in the system are presented. Assessment of the environmental quality of the final product and the consequent guarantee of its use in the building industry demand that it meets a number of requisites, one of which is that the effluents extracted by water action should be contamination-free, or at least that the concentration of contaminants should be below certain pre-set limits. For this a number of leaching tests must be carried out, such as the Netherlands Leaching Test . PMID- 11942704 TI - Waste management options in southern Europe using field and experimental data. AB - The applicability of the Waste Management Hierarchy concept that appeared to be an essential element in current national environmental policies was investigated in the case of a region in Southern Europe. The waste generation profile that determines the appropriateness of different waste management options was created after a 1-year municipal waste sampling investigation conducted in the Municipality of Pilea in Northern Greece. The paper describes the results of (1) the sampling method, which was conducted four times during 1 year (once per season) in selected areas of the city, (2) the qualitative analysis of the collected samples and (3) the waste treatment, which consisted of drying, grinding, calorific value measurement, incineration and chemical analysis of the collected samples. Comparative analysis between the above mentioned data and on past data derived from investigations conducted in other Greek regions with similar characteristics to those of Pilea were used to identify and discuss future trends in the composition of generated waste over time. An analysis of the current waste management status in Greece as well as the feasibility of implementing a comprehensive management approach is assessed taking into account guidelines set worldwide to promote renewable energy sources use. It is concluded that recycling, perhaps the most positively received of all waste management practices, is going to be an essential part of contemporary waste management strategies, composting can play an important role, while incineration seems to be a conditionally feasible solution. PMID- 11942705 TI - A laboratory pilot study of thermal decontamination of soils polluted by PCBs. Comparison with thermogravimetric analysis. AB - Real soils contaminated with a mixture of polychlorinated biphenyls were thermally decontaminated in a laboratory scale thermal desorption apparatus, under different operating conditions including two operating pressures (P= 0.01 and 0.1 MPa), different sample masses (80-320 g), carrier gas flow rates (0-30 280 Nl h(-1)) and two initial contamination levels. A standard European soil artificially contaminated with 4-chlorobiphenyl was decontaminated using a thermogravimetric analyser (TGA). The same parametric study as cited earlier was performed. With both techniques, the extent of decontamination was studied as a function of temperature during the heating programs (2-3 degrees C min(-1) for the thermal desorption apparatus; 5 degrees C min(-1) for TGA). Only a few differences were noticed between the two techniques. The decontamination starts when the melting temperature of the contaminant is reached (30 degrees C) and is complete before 350 degrees C. Thermogravimetric analysis which does not necessitate any chemical analysis appears to be a very attractive technique to investigate the feasibility of a decontamination and to quickly determine the best operating conditions. PMID- 11942706 TI - Bioreactor landfills: experimental and field results. AB - Bioreactor landfills allow a more active landfill management that recognizes the biological, chemical and physical processes involved in a landfill environment. This paper presents the results of an experimental study carried out to determine the effect of solid waste size, leachate recirculation and nutrient balance on the rate of municipal solid waste (MSW) biodegradation. Higher rates of MSW biodegradation eventually cause a reduction of the contaminant life span of the landfill and decrease in the cost of long term monitoring. The study indicated that the smaller the size of the MSW the faster the biodegradation rate of the waste. In addition, the paper presents the results of leachate recirculation on solid waste biodegradation in a full-scale landfill site, which is located in Nepean, Ontario, Canada. The leachate was recirculated into the landfilled solid waste for 8 years through infiltration lagoons. Similar results to those obtained in the laboratory scale experiments were noted. The average pH of the leachate in the early stages of recirculation was on the acidic range of the pH scale, however, the pH value was in the range of 7-8 after 2 years of leachate recirculation. The concentration of chloride remained fairly constant at about 1000 mg/l during the leachate recirculation period. A decreasing trend of the organic load, measured as biological oxygen demand and chemical oxygen demand, was observed. Recovery of landfill air space was also noted because of the enhanced subsidence and decomposition of the solid waste. PMID- 11942708 TI - Recovery of nickel oxide from spent catalyst. AB - This study investigates the possibility of recovering nickel from the spent catalyst (NiO/Al2O3) resulting from the steam reforming process to produce water gas (H2/H2O) in many industries. In the extraction process, nickel is recovered as sulfate using sulfuric acid as a solvent. The considered parameters affecting nickel recovery were acid concentration, temperature and time of digestion solid:liquid ratio, particle size and stirring rate. Nickel was to be directly recovered as a sulfate salt by direct crystallization method. The conversion was 99% at 50% sulfuric acid concentration, solid: liquid ratio (1:12) by weight, particle size less than 500 micron for more than 5 h and 800 rpm at 100 degrees C. PMID- 11942707 TI - Wet oxidative regeneration of activated carbon loaded with reactive dye. AB - Wet Oxidative Regeneration (WOR) of powdered activated carbon (PAC) and granular activated carbon (GAC) loaded with the reactive dyes, namely chemictive brilliant blue R and cibacron turquoise blue G, was studied. Attempts were made to regenerate the loaded carbons designated now as spent carbon. A slurry (10% w/v) of spent carbon in distilled water was oxidized by wet oxidation in the temperature range of 150-250 degrees C using oxygen partial pressures between 0.69-1.38 MPa in an 1 1 SS 316 autoclave. The percent regeneration was determined from a ratio, X(RC)/X(VC), corresponding to an equilibrium adsorption capacity of regenerated carbon/equilibrium adsorption capacity of virgin carbon from an initial adsorption period of 3 h. It was observed that the regeneration mainly occurred due to the oxidation of the adsorbates taking place on the surface of carbon. It was possible to regenerate the spent GAC and PAC to the extent of more than 98% (approximately X(RC)/X(VC) > 0.98) by wet oxidation. After four consecutive cycles of adsorption and regeneration using the same stocks of GAC, carbon weight loss observed at 200 degrees C was about 40%. SEM studies of the regenerated carbon showed widening of the pores and loss of structure between the adjacent pores as compared with the virgin carbon. PAC was found to be more suitable as compared with GAC for the adsorption and wet oxidative regeneration processes to treat the aqueous solution containing lower concentration of unhydrolyzed reactive dye. The suitability of wet oxidative regeneration is demonstrated at a bench scale to treat the synthetic reactive dye solution. PMID- 11942709 TI - Removal of polychlorinated biphenyls from capacitors and pressure-sensitive paper by vacuum thermal recycling. AB - This paper describes a new vacuum thermal recycling (VTR) method for treating discarded polychlorinated biphenyls (PCB)-contaminated capacitors and pressure sensitive paper. Based on results demonstrating an exceptionally high PCB separation efficiency and recovery ratio from capacitors containing high concentration PCBs, i.e. > 99.9%, respectively, the presented VTR method is verified to effectively remove PCBs. In addition, associated safety aspects of the employed procedure were confirmed. PMID- 11942710 TI - The long term behaviour of CFBC ash-water systems. AB - Circulating fluidized bed combustion (CFBC) ashes in contact with water, whether in liquid or vapour form, exhibit considerable chemical activity, which can continue for more than a year. From the point of view of the setting of CFBC ash water mixtures and their behaviour, the formation of gypsum and of ettringite are important. The crystallisation of ettringite is associated with expansion of the set mass and absorption of water into the solids, but the ash-water system as a whole shrinks. The extent of ettringite formation is reduced if carbonation of free lime can take place. There is little carbonation if the set solids are covered by water but if contact with air is not restricted, the ettringite formed earlier slowly decomposes, its place being taken by calcite and gypsum. The fate of the aluminium from decomposed ettringite is uncertain; no new crystalline Al compounds are found. It appears that ultimately most analytical lime present can become carbonated. This is environmentally beneficial since it reduces chemical activity in the landfill, and reduces the amount of CO2 released due to limestone use. The main features of the behaviour of CFBC ash-water mixtures can be reproduced in model systems (CaO-Al2O3-CaSO4-H2O) but that of real ashes is complicated by the presence of silicates and possible interactions between silica and the strongly alkaline aqueous medium. Also, aluminates present in the ash are probably more active with respect to ettringite formation than pure aluminum hydroxide. PMID- 11942711 TI - 2001 SOPHE presidential address. The power of partnerships. Society for Public Health Education. PMID- 11942712 TI - Clustering women's health behaviors. AB - This study attempts to characterize health lifestyles by subgrouping women with similar behavior patterns. Data on background, health behaviors, and perceptions were collected via phone interview from 1,075 Israeli women aged 50 to 74. From a cluster analysis conducted on health behaviors, three clusters emerged: a "health promoting" cluster (44.1%), women adhering to recommended behaviors; an "inactive" cluster (40.3%), women engaging in neither health-promoting nor compromising behaviors; and an "ambivalent" cluster (15.4%), women engaging somewhat in both health-promoting and compromising behaviors. Clustering was cross-tabulated by demographic and perceptual variables, further validating the subgrouping. The cluster solution was also validated by predicting another health behavior (mammography screening) for which there was an external validating source. Findings are discussed in comparison to published cluster solutions, culminating in suggestions for intervention alternatives. The concept of lifestyle was deemed appropriate to summarize the clustering of these behavioral, perceptual, and structural variables. PMID- 11942713 TI - Analyzing the contribution of community change to population health outcomes in an adolescent pregnancy prevention initiative. AB - Few evaluations of community initiatives have established a link between intermediate outcomes, such as community or systems change, and more distant population-level health outcomes (e.g., estimated rates of employment or adolescent pregnancy). This article describes an analysis of the contribution of community changes facilitated by a community health initiative to prevent adolescent pregnancy to the population-level outcome of birth rates for teens. The authors examine a hypothesis that this link might be expected when community changes are of greater amount, intensity, duration, and exposure. The results showed reductions in birth rates in Target Area A, where there was a greater concentration of community changes and a slight increase where there were far fewer changes. This report provides a method for describing empirically the contribution of environmental change to more distant population-level outcomes. PMID- 11942714 TI - Parenting practices and adolescent risky driving: a three-month prospective study. AB - This study examined relations between risky driving, parenting, and deviance, and the stability of risky driving over time. Two hundred and sixty-one licensed adolescents completed telephone interviews about risky driving, parenting practices, and orientations toward deviance at baseline and about risky driving at follow-up 3 months later. The results indicated that risky driving at follow up was predicted by risky driving at baseline, parental restrictions on driving, and sensation seeking. In addition, risky driving was stable within 80% of teens. When compared with adolescents with low risky driving over time (n = 129), adolescents with high risky driving over time (n = 79) were 3 times more likely to report low parental monitoring, 2 times more likely to report low parental restrictions, and almost 5 times more likely to report high deviance acceptance. The results suggest that high levels of risky driving are related to parenting. PMID- 11942715 TI - Improving adherence to regular pap smear screening among Asian women: a population-based study in Singapore. AB - This study addresses the issue of adherence to regular screening for cervical cancer in Asian women and factors that would promote this. A community-based cross-sectional survey was carried out among 726 women (30-59 years) in Singapore. Although the overall screening prevalence was 62.4%, only 42% reported having regular smears. Women in the irregular group were more likely to have a Pap smear as part of postnatal or family planning visits, whereas the primary reason for attendance among the regular group was likely to be screening or a health checkup. Chief barriers to regular screening were low perceived susceptibility (46.5%) and low accessibility (37.1%). One third of women who have ever had a smear do not return for regular screening. Appropriate windows of opportunity in contact between women and the health care services should be used to convey the importance of adherence to screening. PMID- 11942717 TI - Qualitative process evaluation of urban community work: a preliminary view. AB - Community-based and participatory research have become significant activities during the past 15 years as public health practitioners and researchers have sought new ways to provide effective disease prevention and health promotion programs. It is also important that more examples of evaluation schemes be contributed for field testing. The process evaluation model offered here was based on an eclectic literature search because the evaluators did not know what directions this Centers for Disease Control and Prevention (CDC)-funded Urban Research Center would take and wanted to be thorough. Participatory action research using predominantly qualitative methods provided a research approach congruent with the capacity-building and power-sharing principles of the center. Seattle Partners for Healthy Communities is not a traditional community project, acting primarily as a broker for expertise and community needs. However, it has been successful in supporting and evaluating community health projects. PMID- 11942716 TI - Condom use and correlates of African American adolescent females' infrequent communication with sex partners about preventing sexually transmitted diseases and pregnancy. AB - This study of 522 African American female adolescents, ages 14 to 18, investigated associations between condom use and infrequently communicating with sex partners about sexually transmitted diseases (STDs) and pregnancy prevention. Correlates of infrequent communication were identified. Sexually active adolescents were recruited from schools and adolescent medicine clinics in low income neighborhoods of Birmingham, Alabama. Adolescents completed a self administered survey and face-to-face interview. Communication frequency was assessed using a five-item scale. Infrequent communication was significantly associated with lower odds of condom use. Multivariate correlates of infrequent communication were less frequent communication with parents about STD/pregnancy prevention, recent sex with a nonsteady partner, low perceived ability to negotiate condom use and fear of this negotiation, and low motivation to use condoms. Given the importance of partner communication in promoting safer sex behaviors, STD and pregnancy prevention programs may benefit adolescents by addressing the identified psychosocial correlates of infrequent communication with their partners. PMID- 11942718 TI - Health promotion attitudes and strategies in older adults. AB - The purpose of this study was to describe older adults' attitudes and beliefs regarding wellness, self-care, and participation in health promotion activities. Six focus groups were conducted with 45 men and women aged 62 to 91. The White Crane Model of Healthy Lives for Older Adults was developed based on the focus group themes. Being healthy incorporates multiple components, including functional independence, self-care and management of illness, positive outlook, and personal growth and social contribution. Being healthy is determined by self evaluation in the context of chronic illness or disability and the social and/or physical environment. Strategies for maintaining health and motivation for healthy behavior are identified. Socialization and social support are central to participation in programs, and interpersonal engagement is particularly important. Flexibility in choice and structure of programs contribute to seniors' sense of control over their health. Programs should be challenging and tailored to older adults' needs and expectations. PMID- 11942719 TI - An equivalent current source model and laplacian weighted minimum norm current estimates of brain electrical activity. AB - We have developed a method for estimating the three-dimensional distribution of equivalent current sources inside the brain from scalp potentials. Laplacian weighted minimum norm algorithm has been used in the present study to estimate the inverse solutions. A three-concentric-sphere inhomogeneous head model was used to represent the head volume conductor. A closed-form solution of the electrical potential over the scalp and inside the brain due to a point current source was developed for the three-concentric-sphere inhomogeneous head model. Computer simulation studies were conducted to validate the proposed equivalent current source imaging. Assuming source configurations as either multiple dipoles or point current sources/sinks, in computer simulations we used our method to reconstruct these sources, and compared with the equivalent dipole source imaging. Human experimental studies were also conducted and the equivalent current source imaging was performed on the visual evoked potential data. These results highlight the advantages of the equivalent current source imaging and suggest that it may become an alternative approach to imaging spatially distributed current sources-sinks in the brain and other organ systems. PMID- 11942720 TI - A new paradigm for the closed-loop intraoperative administration of analgesics in humans. AB - We present a new paradigm for the closed-loop administration of analgesics during general anesthesia. The manipulated variable in the control system is the infusion rate of the opiate alfentanil, administered intravenously through a computer-controlled infusion pump (CCIP). The outputs to be controlled are the patient's mean arterial pressure (MAP) and the drug concentration in the plasma. Maintaining MAP within appropriate ranges provides optimal treatment of the patient's reactions to surgical stimuli. Maintaining plasma drug concentrations close to a reference value specified by the anesthesiologist allows to titrate analgesic administration to qualitative clinical end-points of insufficient analgesia. MAP is acquired invasively through a catheter cannula. Since plasma drug concentrations cannot be measured on-line, they are estimated via a pharmacokinetic model. We describe an explicit model-predictive controller which achieves the above-mentioned objectives. An upper constraint on drug concentrations is maintained to avoid overdosing. Constraints on the MAP are introduced to trigger a prompt controller reaction during hypertensive and hypotensive periods. Measurement artifacts in the MAP signal are rejected to prevent harmful misbehavior of the controller. We discuss the results of the clinical validation of the controller on humans. PMID- 11942721 TI - A statistical model for interpreting computerized dynamic posturography data. AB - Computerized dynamic posturography (CDP) is widely used for assessment of altered balance control. CDP trials are quantified using the equilibrium score (ES), which ranges from zero to 100, as a decreasing function of peak sway angle. The problem of how best to model and analyze ESs from a controlled study is considered. The ES often exhibits a skewed distribution in repeated trials, which can lead to incorrect inference when applying standard regression or analysis of variance models. Furthermore, CDP trials are terminated when a patient loses balance. In these situations, the ES is not observable, but is assigned the lowest possible score--zero. As a result, the response variable has a mixed discrete-continuous distribution, further compromising inference obtained by standard statistical methods. Here, we develop alternative methodology for analyzing ESs under a stochastic model extending the ES to a continuous latent random variable that always exists, but is unobserved in the event of a fall. Loss of balance occurs conditionally, with probability depending on the realized latent ES. After fitting the model by a form of quasi-maximum-likelihood, one may perform statistical inference to assess the effects of explanatory variables. An example is provided, using data from the NIH/NIA Baltimore Longitudinal Study on Aging. PMID- 11942722 TI - A quantitative evaluation of the magnetic field generated by a CA3 pyramidal cell at EPSP and action potential stages. AB - We evaluate quantitatively which behavioral stage dominantly generates magnetic field adjacent to a CA3 pyramidal cell by using a compartmental model with dendrites and an axon. Generally speaking, there are four stages in the potential behavior, i.e., excitatory and inhibitory postsynaptic potential, firing action potential, bursting action potential, if any, and after hyperpolarization potential stages. Calculated magnetic field also consists of corresponding four stages. We find, first, the dominant origin of the peaks of the magnetic field is counter propagating pulses at the firing and bursting stage at basal and apical dendrites. Second, the amplitude of the magnetic field changes to a great extent by the cancellation timing of the apical- and basal-originating fields depending on the calcium ionic channel spikes. Third, the field generated by the current flowing through the axon is significant enough when the temporal resolution of the measurement system becomes high. The results predict that the magnetic-field waveform measured in physiological experiments represents the dendritic configurations, channel density distributions, and bursting characteristics. These facts enable new investigations of neuronal activities in more detail through the observation of the magnetic-field waveform. PMID- 11942723 TI - Modeling of corneal surfaces with radial polynomials. AB - We consider analytical modeling of the anterior corneal surface with a set of orthogonal basis functions that are a product of radial polynomials and angular functions. Several candidate basis functions were chosen from the repertoire of functions that are orthogonal in the unit circle and invariant in form with respect to rotation about the origin. In particular, it is shown that a set of functions that is referred herein as Bhatia-Wolf polynomials, represents a better and more robust alternative for modeling corneal elevation data than traditionally used Zernike polynomials. Examples of modeling corneal elevation are given for normal corneas and for abnormal corneas with significant distortion. PMID- 11942724 TI - Parameter estimation of human nerve C-fibers using matched filtering and multiple hypothesis tracking. AB - We describe how multiple-target tracking may be used to estimate conduction velocity changes and recovery constants of human nerve C-fibers. These parameters discriminate different types of C-fibers and pursuing this may promote new insights into differential properties of nerve fiber membranes. Action potentials (APs) were recorded from C-fibers in the peroneal nerve of awake human subjects. The APs were detected by a matched filter constituting a maximum-likelihood constant false-alarm rate detector. Using the multiple-hypothesis tracking method and Kalman filtering, the detected APs (targets) in each trace (scan) were associated to individual nerve fibers (tracks) by their typical conduction latencies in response to electrical stimulation. The measurements were one dimensional (range only) and the APs were spaced in time with intersecting trajectories. In general, the AP amplitude of each C-fiber differed for different fibers. Amplitude estimation was therefore incorporated into the tracking algorithm to improve the performance. The target trajectory was modeled as an exponential decay with three unknowns. These parameters were estimated iteratively by applying the simplex method on the parameters that enter nonlinearly and the least squares method on the parameters that enter linearly. PMID- 11942725 TI - Interscale wavelet maximum-a fine to coarse algorithm for wavelet analysis of the EMG interference pattern. AB - A method has been developed, interscale wavelet maximum (ISWM), for characterising the electromyogram (EMG) interference pattern to assist in the diagnosis of neuromuscular disease. EMG signals are decomposed with the redundant dyadic wavelet transform and wavelet maxima (WM) are found. Thresholding methods are applied to remove WM due to noise and background activity. An efficient fine to-coarse algorithm identifies the WM tree structure for the motor unit action potential rising edges. The WM for each tree are summed at each scale; the largest value is the ISWM. Highly significant differences in ISWM values have been found between healthy, myopathic, and neuropathic subjects that could make the technique a useful diagnostic tool. PMID- 11942726 TI - Cardiac artifact subspace identification and elimination in cognitive MEG data using time-delayed decorrelation. AB - To reduce physiological artifacts in magnetoencephalographic (MEG) and electroencephalographic recordings, a number of methods have been applied in the past such as principal component analysis, signal-space projection, regression using secondary information, and independent component analysis. This method has become popular as it does not have constraints such as orthogonality between artifact and signal or the need for a priori information. Applying the time delayed decorrelation algorithm to raw data from a visual stimulation MEG experiment, we show that several of the independent components can be attributed to the cardiac artifact. Calculating an average cardiac activity shows that physiologically different excitation states of the heart produce similar field distributions in the MEG sensor system. This is equivalent to differing spectral properties of cardiac field distributions in the raw data. As a consequence, the algorithm combines, e.g., the R peak and the T wave of the cardiac cycle into a single component and the one-to-one assignment of each independent component with a physiological source is not justified in this case. To improve the signal quality of visually evoked fields, the multidimensional cardiac artifact subspace is suppressed from the data. To assess the preservation of the evoked signal after artifact suppression, a geometrical and a temporal measure are introduced. The suppression of cardiac and alpha wave artifacts allows, in our experimental setting, the reduction of the number of epochs to one half while preserving the visually evoked signal. PMID- 11942727 TI - An efficient coding algorithm for the compression of ECG signals using the wavelet transform. AB - A wavelet-based electrocardiogram (ECG) data compression algorithm is proposed in this paper. The ECG signal is first preprocessed, the discrete wavelet transform (DWT) is then applied to the preprocessed signal. Preprocessing guarantees that the magnitudes of the wavelet coefficients be less than one, and reduces the reconstruction errors near both ends of the compressed signal. The DWT coefficients are divided into three groups, each group is thresholded using a threshold based on a desired energy packing efficiency. A binary significance map is then generated by scanning the wavelet decomposition coefficients and outputting a binary one if the scanned coefficient is significant, and a binary zero if it is insignificant. Compression is achieved by 1) using a variable length code based on run length encoding to compress the significance map and 2) using direct binary representation for representing the significant coefficients. The ability of the coding algorithm to compress ECG signals is investigated, the results were obtained by compressing and decompressing the test signals. The proposed algorithm is compared with direct-based and wavelet-based compression algorithms and showed superior performance. A compression ratio of 24:1 was achieved for MIT-BIH record 117 with a percent root mean square difference as low as 1.08%. PMID- 11942728 TI - Feature analysis and centromere segmentation of human chromosome images using an iterative fuzzy algorithm. AB - Classification of homologous chromosomes is essential to advanced studies of cancer genetics. Centromere intensities are believed to be an important differentiating feature between homologs. Therefore, segmentation of centromeres is a major step toward the realization of homolog classification. This paper describes an iterative fuzzy algorithm which successfully segments centromeres from images of human chromosomes prepared using fluorescence in-situ hybridization technique. The algorithm is based on assigning a fuzzy membership value to each pixel in the centromere image. An iterative algorithm then updates and minimizes a defined error function. Chromosome 22, a highly heteromorphic chromosome, is used to verify the centromere segmentation method. Homologs of this chromosome are classified based on their segmented centromere intensities as well as their morphological differences. The classification results of these two methods agree completely and are used to validate our developed algorithm. PMID- 11942729 TI - Cascaded differential and wavelet compression of chromosome images. AB - This paper proposes a new method for chromosome image compression based on an important characteristic of these images: the regions of interest (ROIs) to cytogeneticists for evaluation and diagnosis are well determined and segmented. Such information is utilized to advantage in our compression algorithm, which combines lossless compression of chromosome ROIs with lossy-to-lossless coding of the remaining image parts. This is accomplished by first performing a differential operation on chromosome ROIs for decorrelation, followed by critically sampled integer wavelet transforms on these regions and the remaining image parts. The well-known set partitioning in hierarchical trees (SPIHT) (Said and Perlman, 1996) algorithm is modified to generate separate embedded bit streams for both chromosome ROIs and the rest of the image that allow continuous lossy-to-lossless compression of both (although lossless compression of the former is commonly used in practice). Experiments on two sets of sample chromosome spread and karyotype images indicate that the proposed approach significantly outperforms current compression techniques used in commercial karyotyping systems and JPEG-2000 compression, which does not provide the desirable support for lossless compression of arbitrary ROIs. PMID- 11942730 TI - An automated neck flexibility tester. AB - We investigate automation and control options for the neck flexibility tester (NFT) (P. McClure et al., 1998), a device originally used to measure the flexibility of the human cervical spine. The motivation is to lay the foundations for design and implementation of investigative devices that would allow studies of mechanical properties of the spine under repetitive dynamic loading. We derive the equations of motion for an automated NFT (ANFT) using a Lagrangian formulation. These equations, which represent a simplified first-order model of the dynamics, are used to simulate the ANFT using the software package Simulink. Two control schemes are examined: proportional plus integral plus derivative (PID) control and dynamic inversion. Both are simulated for setpoint and tracking control. It appears that PID control is preferred due to its simplicity of design and relative insensitivity to the dynamic model of the ANFT. PMID- 11942731 TI - Tissue reactions evoked by porous and plane surfaces made out of silicon and titanium. AB - Square-shaped silicon or titanium implants with plane or porous surfaces surrounded by a rim of silicone were implanted in the rat abdominal wall for evaluation of the tissue response after one, six, or 12 weeks. Cell damage was identified as increased membrane permeability using fluorescence microscopy by injection of propidium iodide prior to the killing of the rats. Capsule thickness and immunohistochemical quantification of macrophages were used as a further measure of the foreign-body reaction. There were no significant differences in capsular cell densities for macrophages, total cells (macrophages, fibroblasts, and other cells), or necrotic cells at the different time points for the four surfaces studied. However, significant differences in the kinetics of the response were found between plane surfaces compared with porous ones. Both types of plane surfaces developed a significant increase in capsule thickness over time in contrast to the porous implants. Porous silicon displayed a significant decrease in total cells in the reactive capsule over time. Furthermore, porous silicon and titanium surfaces displayed a significant decrease in total cell numbers at the implant interface between six and 12 weeks. The present study demonstrated that implanted silicon elicited soft-tissue reactions comparable to that of titanium. PMID- 11942732 TI - A feasibility study for electrical impedance tomography as a means to monitor tissue electroporation for molecular medicine. AB - Molecular medicine involves the introduction of macromolecules, such as drugs or gene constructs, into specific cells of the body. Electroporation, which uses electric pulses to permeate cell membranes, is a method for achieving this. However, as with other molecular medicine procedures, it lacks a real-time mechanism to detect and control which cells have been affected. We propose and demonstrate, via computer simulation, that electrical impedance tomography has the potential for detecting and imaging electroporation of cells in tissue in real-time, thereby providing feedback for controlling electroporation. PMID- 11942733 TI - Limited joint mobility in selected hand and foot joints in patients with type 1 diabetes mellitus: a methodology comparison. AB - OBJECTIVES: First to determine the range of motion (ROM) of selected foot and hand joints with a goniometer, 2.) to determine joint limitation by prayer sign and 3.) to compare both methods used. METHODS: Maximal active ROM was measured by goniometry (Method 1) in 50 patients with Type 1 diabetes and in 44 healthy controls, respectively. The lower limits for normal ranges were defined as mean minus 2 SD. To elicit the prayer sign (Method 2) subjects were asked to put their hands together in a praying position with the fingers fanned. The number of missing joint contacts between the fingers was counted and correlated to ROM. RESULTS: The ROM of all joints measured, except that of the thumb, was significantly smaller in diabetic patients than in control subjects. The degree of the prayer sign was well correlated to the ROM of most hand joints, but was solely correlated in feet to ankle ROM. In diabetic patients the prevalence of limited joint mobility (LJM) measured by goniometry ranged from 2 to 20% in hand joints and from 10 to 14% in foot joints. By the prayer sign 33% of the diabetic patients had LJM. CONCLUSION: With both methods, diabetic patients had more limitations in joint motion than control subjects, as was their prevalence of LJM higher. Since the prayer sign was correlated to exactly measured ROMs, we would suggest using the prayer sign as a simple clinical indicator for LJM in diabetic subjects. PMID- 11942734 TI - Evaluation of oxidative stress in diabetic patients after supplementation with a standardised red orange extract. AB - Diabetes mellitus is associated with a high oxidative stress level, resulting from an imbalance between free radicals or reactive oxygen species production and the antioxidant systems. Inhibition of these oxidative processes by co-adjuvant therapy could therefore prevent, or at least delay, the onset and/or the development of long-term diabetic complications. Dietary supplementation with plant biophenols may be a successful strategy to decrease this risk of pathological complications. The Red Orange Complex (ROC) is a standardized red orange extract containing, as its main active principles, phenolic compounds (anthocyanins, flavanones and hydroxycinnamic acids) as well as ascorbic acid. The aim of the present preliminary study was to evaluate the effects of short term (2 mo) dietary supplementation with ROC (50 mg/d, orally) on some serum non invasive biomarkers of oxidative stress (total antioxidant status, or TAS, levels of thiol groups and levels of free radicals) in a group of 33 patients with Type 2 diabetes, in comparison with a group of 28 healthy volunteers. The results obtained demonstrate that in diabetic patients supplementation with ROC can improve blood levels of thiol groups on proteins (an indirect measurement of glutathione activity in serum); furthermore, it can elicit a marked decrease in serum free radical levels, in patients with high blood oxidative stress status. However, ROC supplementation appeared unable to modify serum TAS. Finally, the glycemic profile remained stable during the study period in all subjects, and no unpleasant side effects were reported. In conclusion, the treatment of diabetic patients with ROC might be of therapeutic benefit in order to protect against diabetes complications that are partially due to uncontrolled lipid oxidation. D PMID- 11942735 TI - Body composition and pulmonary function in obese type 2 diabetic women. AB - Most patients with Type 2 diabetes mellitus (T2DM) are overweight and their fat distribution pattern shows more truncal and less peripheral subcutaneous fat. Obesity also influences the respiratory system by mechanical effects on the diaphragm and chest wall depending on the distribution and size of excess adipose tissues. To determine if the impairment of pulmonary function in T2DM is associated with truncal fat distribution, we performed dynamic spirometric tests and body composition analysis, by dual X-ray absorptiometry, in 12 non-smoker mildly obese T2DM women and in an equal number of control normoglycaemic females, matched for age, body mass index and smoking habits. The forced vital capacity and forced expiratory volume in one second for T2DM women (2.67 +/- 0.30 and 2.15 +/- 0.20 1, respectively) were significantly (p < 0.05) lower than for control women (3.22 +/- 0.30 and 2.58 +/- 0.31 1, respectively). However, trunk fat mass and trunk lean body mass for T2DM women (16.68 +/- 2.31 and 20.87 +/- 1.91 kg, respectively) were comparable with those for controls (16.46 +/- 2.54 and 19.65 +/- 3.42 kg, respectively). Thus, we deduce that pulmonary function impairment in T2DM obese women is not associated with truncal fat mass deposition or with lean mass depletion. PMID- 11942736 TI - Neurophysiology of micturition and pharmacotherapy of lower urinary tract dysfunction. PMID- 11942737 TI - Diabetic cystopathy. PMID- 11942738 TI - Modulation of penile smooth muscle cells contractility. PMID- 11942739 TI - Experience with sildenafil in diabetes. PMID- 11942740 TI - Intracavernosal injection therapy and surgical therapy in diabetic patients with erectile dysfunction. PMID- 11942741 TI - Management of erectile dysfunction in diabetic patients. PMID- 11942742 TI - Treatment with the insulin analogue lispro in children and adolescents with type 1 diabetes mellitus: evaluation over a 3-year period. AB - The aim of the present study was to evaluate the degree of metabolic control obtained with the use of the insulin analogue lispro compared to the previous regimen with classical regular insulin in children and adolescents with Type 1 diabetes mellitus. HbA1c, lipid metabolism, body mass index (BMI), frequency of severe hypoglycaemia, carbohydrate intake, total daily insulin requirements and its distribution during the day were analysed in 44 diabetics patients (57% males and 43% females) throughout a 3-yr period. The mean age of the patients at the beginning of the study was 15.6 +/- 2.7 yr with a mean duration of the disease of 8.01 +/- 3.4 yr. All data were evaluated for the year before the change of treatment, and 1 yr (44 patients), 2 yr (19 patients) and 3 yr (13 patients) after the change. HbA1c levels did not significantly change (6.6 +/- 1.1% with regular insulin, 6.32 +/- 1.05% in the 1st year with lispro, 6.6 +/- 1.1% in the 2nd yr with lispro, 6.33 +/- 0.9% in the 3rd yr with lispro). However, significant differences (p = 0.03) were found after 3 yr of treatment in those patients who changed to insulin lispro therapy due to a bad glycaemic control. The total daily insulin dose (U/kg/d) remained unchanged. The total short acting/intermediate-acting insulin ratio significantly decreased (45.9 +/- 0.1% regular insulin; 37.2 +/- 0.1% 1st yr lispro (p < 0.001); 33.6 +/- 0.1% 2nd yr lispro (p < 0.05); 35.5 +/- 0.1% 3rd yr lispro (p < 0.05). BMI and lipid profile remained unchanged. The self-reported daily carbohydrate in take significantly decreased due to a reduction of snacks. Total number of episodes of severe hypoglycaemia did not change significantly. In conclusion insulin lispro treatment did not modify the daily insulin dose, but reduced the short acting/intermediate acting insulin ratio. The metabolic control remained unchanged. The number of patients reporting severe hypoglycaemia was similar despite the treatment schedule. After this 3-yr duration trial all patients decided to continue the treatment with lispro insulin. PMID- 11942743 TI - Measuring integrated pest management programs for public buildings. AB - Integrated pest management (IPM) tends to be perceived by different stakeholder groups either as a methodology for effective pest control or as an ideology of responsible environmental stewardship. The IPM process has never been subjected to a rigorous empirical test as a control methodology in buildings; published studies have either tested isolated program components or have presented uncontrolled, sequential descriptions of IPM replacing traditional pest control service procedures. Because ideological measurement is simpler, cheaper, and more relevant than methodological testing to evaluate structural IPM performance in the public sector, data on pesticide use/risk and customer satisfaction, rather than control efficacy, are used by the General Services Administration (GSA) IPM program to demonstrate success compatible with Government Performance and Results Act (GPRA) guidelines. Implementation of IPM in 1989 resulted in significant decreases both in quantities of insecticide applied indoors and requests for pest control service by building occupants throughout the first decade of the program. Although these results do not provide an empirical test of structural IPM methodological superiority as a means of reducing pest populations, they indicate that replacing sprayed insecticide formulations with baits and using client reporting as the primary pest surveillance method can successfully achieve the policy goals of a large-scale IPM program for public buildings. PMID- 11942744 TI - Incidence, symptoms, and intensity of damage by three coffee stem borers (Coleoptera: Cerambycidae) in South Yunan, China. AB - Sampling studies were conducted in coffee plantations in South Yunan to assess the incidence, symptoms, and intensity of damage by three stem borers: Xylotrechus quadripes (Chevrolat), Acalolepta cervina (Hope), and Bacchisa sp. near pallidiventris (Thomson). Of 5,690 plants sampled in eight plantations, 440 were infested with A. cervina, 63 with X. quadripes, and three with B. pallidiventris. Plants 5-7 yr old were 10 times more heavily infested with X. quadripes than 3- to 4-yr-old plants, whereas both age groups of plants had similar levels of infestation with A. cervina. Larval galleries of the three borer species markedly differ: A. cervina and B. pallidiventris larvae develop in subcortical galleries in the main stem (A. cervina) and lateral branches (B. pallidiventris), whereas larval galleries of X. quadripes intermittently punctuate and transverse the xylem of main stems or lateral branches. Significantly more plant tissue was damaged in stems infested with X. quadripes than in those infested with A. cervina or B. pallidiventris. Stems infested with A. cervina or B. pallidiventris generally had only one or a few pupation chambers, whereas stems infested with X. quadripes contained numerous chambers. Quantitative and qualitative data collected through this study provide farmers with diagnostic tools to determine which borer species infested coffee plants. Comparison of life history traits and intensity of damage for the three borer species indicates that X. quadripes is the most severe pest of coffee in Yunan, and suggests that populations of X. quadripes have the greatest potential to steadily increase with time. PMID- 11942745 TI - Field trials measuring the effects of ultraviolet-absorbing greenhouse plastic films on insect populations. AB - Field studies were conducted to compare insect population levels in greenhouse crops covered with plastics that block the transmission of UV light in two wavelength ranges. Crops grown in greenhouses under a plastic that blocked UV light at wavelengths of 380 nm and below had lower numbers of aphids and thrips compared with a plastic that blocked UV light at wavelengths of 360 nm and below. This is consistent with the results found for thrips in previous studies using small, completely enclosed tunnels with no plant material. The effects were not as dramatic in the commercial greenhouses, perhaps due to unfiltered light entering through the open sides of those greenhouses. There was no reduction in greenhouse whitefly, Trialeurodes vaporariorum Westwood, populations under the <380 UV-absorbing plastic compared to <360 nm UV-absorbing plastic in these field trials. This is inconsistent with results found in small, completely enclosed tunnels, where sticky traps caught a significantly higher proportion (95 +/- 2%) of released greenhouse whiteflies inside tunnels covered with <360 nm absorbing plastic compared with the <380 nm absorbing plastic. The results of these studies suggest that the type of greenhouse plastic used in a structure can affect population levels of some insect species, and may be useful tools in developing integrated pest management programs for insect management. The design of the greenhouse and amount of unfiltered light that enters the system appear to be important factors in determining the level of effect. PMID- 11942746 TI - Responses of Coptotermes formosanus and Reticulitermes flavipes (Isoptera: Rhinotermitidae) to three types of wood rot fungi cultured on different substrates. AB - This study examined the responses of two termite species, the Formosan subterranean termite, Coptotermes formosanus Shiraki, and the eastern subterranean termite, Reticulitermes flavipes (Kollar), to three types of wood decay fungi: a brown rot fungus, Gloeophyllum trabeum (Persoon: Fries) Murrill; a white rot fungus, Phanerochaete chrysosporium Burdsall; and a litter rot fungus, Marasmiellus troyanus (Murrill) Singer. We also examined the responses of termites to these three types of fungi grown on different substrates. For all three fungal species, both termite species showed a strong preference for fungus infected sawdust over uninfected sawdust. In choice tests, both termite species preferred sawdust infected with either M. troyanus or P. chrysosporium over G. trabeum. However, termites did not show any preference for fungus-infected potato dextrose agar over uninfected potato dextrose agar. Tunneling activity of C. formosanus was greater in sand treated with methanol extracts of fungus-infected sawdust than in sand treated with extracts of uninfected sawdust. Because chemicals in the fungal extracts caused termites to tunnel further into treated sand than untreated sand, these chemicals could potentially be used to direct termite foraging toward bait stations in the field. PMID- 11942747 TI - Linkage analysis of an acetylcholinesterase gene in the house fly Musca domestica (Diptera: Muscidae). AB - Linkage of an acetylcholinesterase (AChE) gene was detected in the house fly, Musca domestica L., by using the backcross method between a strain, aabys, that had a morphological multichromosomal marker on each of the five autosomes and a wild strain, LPR. Both strains were homozygous in this gene, and we used eight single nucleotide polymorphisms (SNPs) between them to distinguish the parental sequences in the backcrossed progeny, two of which resulted in the amino acid substitiutions common to the Drosophila and Aedes AChEs insensitive to organophosphates and carbamates. F, appeared to be a wild phenotype, and the AChE gene was heterozyous of aabys and LPR. In the backcross progeny, 32 (2(5)) phenotypes appeared, and 10 phenotypes with one wild or morphological marker were picked up for genotyping by the SNPs of AChE gene. A combination of the morphological markers and the SNPs revealed that the AChE structural gene is linked to autosome 2 in the house fly. PMID- 11942748 TI - In-season changes in resistance to insecticides in Helicoverpa armigera (Lepidoptera: Noctuidae) in India. AB - Discriminating doses of fenvalerate, cypermethrin, quinalphos, and endosulfan were determined with an insecticide-susceptible Helicoverpa armigera (Hubner) strain. In-season changes in insecticide resistance were monitored with discriminating dose assays at weekly intervals throughout the cropping season for 6 yr from 1993 to 1999 in central India. Resistance to pyrethroids was high throughout all seasons. Resistance to 0.75 microg of quinalphos was consistent, with seasonal averages ranging from 23 to 27% survival over the 6 yr. Resistance to 10.0 microg of endosulfan was moderately high at an average of 40-47% survival during 1993-1994 and in 1997-1998. It was lower in 1996-1997 at 27%, and in 1998 1999 at 33%. The weekly monitoring data for all seasons were pooled and the consolidated 6-yr seasonal average profile indicated that resistance to quinalphos and endosulfan was low during September at 21 and 27% survival, respectively, but increased to 28 and 37% by the end of November. Resistance levels to organophosphates and endosulfan increased during the season, depending on the use of these compounds. At almost all monitoring sites, the within-season changes in quinalphos resistance for all seasons through the study period followed a trend similar to that of endosulfan. The results suggest the possibility of cross-resistance between these compounds. Based on this study and the existing information on cotton pest management, we have developed a "window strategy" for cotton pest management with specific emphasis on the management of insecticides for effective control of H. armigera. This strategy has contributed to improved control at reduced costs in extensive trials. PMID- 11942749 TI - Examination of the F2 screen for rare resistance alleles to Bacillus thuringiensis toxins in the diamondback moth (Lepidoptera: Plutellidae). AB - A synthetic laboratory population of the diamondback moth, Plutella xylostella (L.), was used to test the F2 screen developed for detecting the frequency of rare resistance alleles to Cry1Ac and Cry1C toxins of Bacillus thuringiensis (Bt). Of the 120 single-pair matings set up, 106 produced enough F2 families for screening of Cry1Ac or Cry1C resistance alleles using both transgenic broccoli and an artificial diet overlay assay with a diagnostic dose. When using Bt broccoli plants as the F2 screen method, only one F2 family was detected for Cry1Ac resistance and no family was detected for Cry1C resistance. Six families were detected for either Cry1Ac or Cry1C resistance using the diet assay. The survivors in the diagnostic diet assay were crossed with the resistant individuals to confirm their resistance genotypes. Four F2 families were confirmed to contain one copy of an allele resistant to Cry1Ac in the original single-pairs and four other F2 families contained an allele resistant to Cry1C. Our results suggest that using transgenic plants expressing a high level of a Bt toxin in an F2 screen may underestimate the frequency of resistance alleles with high false negatives, or fail to detect true resistance alleles. The diagnostic diet assay was a better F2 screen method to detect alleles, especially for the Cry1Ac resistance with monogenic inheritance in the diamondback moth. The estimated probabilities of false positives and false negatives were 33 and 1%, respectively, for detecting Cry1Ac resistance at the allele frequency of 0.012 using the diagnostic diet assay. Careful validation of the screening method for each insect-crop system is necessary before the F2 screen can be used to detect rare Bt resistance alleles in field populations. PMID- 11942750 TI - Oviposition on and mining in bolls of Bt and non-Bt cotton by resistant and susceptible pink bollworm (Lepidoptera: Gelechiidae). AB - Transgenic cotton that produces insecticidal crystal protein Cry1Ac of Bacillus thuringiensis (Bt) has been effective in controlling pink bollworm, Pectinophora gossypiella (Saunders). We compared responses to bolls of Bt cotton and non-Bt cotton by adult females and neonates from susceptible and Cry1Ac-resistant strains of pink bollworm. In choice tests on caged cotton plants in the greenhouse, neither susceptible nor resistant females laid fewer eggs on Bt cotton bolls than on non-Bt cotton bolls, indicating that the Bt toxin did not deter oviposition. Multiple regression revealed that the number of eggs laid per boll was negatively associated with boll age and positively associated with boll diameter. Females also laid more eggs per boll on plants with more bolls. The distribution of eggs among bolls of Bt cotton and non-Bt cotton was clumped, indicating that boll quality rather than avoidance of previously laid eggs was a primary factor in oviposition preference. Parallel to the results from oviposition experiments, in laboratory no-choice tests with 10 neonates per boll, the number of entrance holes per boll did not differ between Bt cotton and non-Bt cotton for susceptible and resistant neonates. Also, like females, neonates preferred younger bolls and larger bolls. Thus, acceptance of bolls by females for oviposition and by neonates for mining was affected by boll age and diameter, but not by Bt toxin in bolls. The lack of discrimination between Bt and non-Bt cotton bolls by pink bollworm from susceptible and resistant strains indicates that oviposition and mining initiation are independent of susceptibility to Cry1Ac. PMID- 11942751 TI - Estimating carrying capacity in a newly colonized sand fly Lutzomyia serrana (Diptera: Psychodidae). AB - The phlebotomine sand fly Lutzomyia serrana (Damasceno & Arouck) was mass-reared tinder conditions of varying densities in an effort to improve colony production efficiency. To do this, the experimental carrying capacity of a standard rearing chamber was determined, i.e., the optimum population size in relation to density (individuals per unit of space). Rearing chambers of 100 cm3 were populated with 1-50 L. serrana engorged females and an equal number of males. Laboratory conditions were maintained at 23-26 degrees C and 85-95% RH. The following parameters were recorded for each experimental chamber (three replicates): (1) female mortality without oviposition, (2) number of eggs oviposited and (3) number of adults emerging from the egg cohort. Female mortality began to increase substantially in the 26-female chamber, from 5.7% to 15% and finally reaching 60.2% in the 46-50 female chambers. In the chambers containing 1-20 females, egg number and realized adult progeny increased linearly to reach an asymptote. In the 20-50 female chambers, the number of eggs ranged from 420 to 699, and adult production from 306 to 432. The optimum carrying capacity for the 100-cm3 chambers was 22 +/- 2 females. Beyond this number, auto-regulation was initiated, i.e., female mortality without oviposition increased as the number of females per chamber increased. Total number of eggs and adult production was similar in all chambers containing 20-50 females. In conclusion, for optimizing production of mass reared sand flies, determination of the carrying capacity is essential to optimize use of insectary resources, to avoid loss of valuable potentially ovipositing females, and to increase overall production efficiency. PMID- 11942752 TI - Comparison of Bacillus thuringiensis corn hybrids and insecticide-treated isolines exposed to bivoltine European corn borer (Lepidoptera: Crambidae) in South Dakota. AB - The performances of Bacillus thuringiensis (Bt) corn hybrids and permethrin treated isolines were evaluated in South Dakota from 1997 to 1999. The corn hybrids were exposed to natural seasonal fluctuations of a bivoltine ecotype European corn borer population. Larval injury parameters, grain yields, and gross incomes were quantified during the 3-yr study. The use of either Bt hybrids or whorl-applied permethrin granules resulted in significant yield advantages in only 1 out of 3 yr of performance testing. Under high corn borer pressure in 1997, the performances of the Bt corn hybrids and permethrin-treated isolines were similar, and significantly better than the untreated isolines. However, injury-free corn did not necessarily translate into higher yields in some of the hybrid groups. Grain moisture at harvest, which can result in moisture penalty or dockage, was significantly higher in some Bt corn hybrids. Neither the Bt hybrids nor the permethrin-treated isolines produced significant advantages in yields in 1998 and 1999 when infestations of first-generation corn borer were almost nonexistent and infestations of second-generation European corn borers were moderate. PMID- 11942753 TI - Distribution and sampling of the whiteflies Aleurothrixus floccosus, Dialeurodes citri, and Parabemisia myricae (Homoptera: Aleyrodidae) in citrus in Spain. AB - From 1993 to 1995 data sets were collected from four citrus groves in Valencia, Spain, to determine the distribution patterns of eggs and nymphs of Aleurothrixus floccosus (Maskell), Dialeurodes citri (Ashmead), and Parabemisia myricae (Kuwana) on leaves, and to develop reliable sampling plans for estimating densities of immature whiteflies. A. floccosus showed higher aggregation than the other two species. The dispersion index b from the Taylor power law did not vary between different developing stages for A. floccosus and D. citri, reaching overall values of 1.70 and 1.53, respectively. In P. myricae, b was 1.60 for eggs and N1, and 1.46 for the remaining nymphs. The minimum number of leaves to estimate the population density with a coefficient of variation of 0.25 for densities above 10 immature whiteflies per leaf was 40 for D. citri and P. myricae, and 250 for A. floccosus. Binomial sampling programs for the three species were rejected for pest management purposes due to the high sample sizes required. The enumerative procedure of counting the number of insects per leaf appears to be the most suitable method for D. citri and P. myricae. For A. floccosus an index of occupation (from 0 to 10) linearly related to the proportion of the leaf undersurface occupied by this insect was found to be reliable and time-saving. Examining 150 leaves with this index achieves the desired relative variation level of 0.25 for most population densities usually found in commercial groves. PMID- 11942754 TI - Susceptibility of the bruchid Callosobruchus maculatus (Coleoptera: Bruchidae) and its parasitoid Dinarmus basalis (Hymenoptera: Pteromalidae) to three essential oils. AB - The bruchid Callosobruchus maculatus (F.) causes major losses during the storage of seeds of Vigna unguiculata (Walp.) in West Africa. An endemic parasitoid, the pteromalid Dinarmus basalis (Rond.) reduces the increase in bruchid populations in stores and could be used for biological control. African farmers often introduce essential oils into granaries at harvest time. In Togo, essential oils were extracted from two Gramineae, Cymbopogon nardus (L.) and Cymbopogon schoenanthus (L.) and from a Lamiaceae, Ocimum basilicum (L.). The major components of these essential oils were citronellal in C. nardus, carene-2 and piperitone in C. schoenanthus and estragol in O. basilicum. Cymbopogon schoenanthus was the most toxic oil for C. maculatus adults. D. basalis adults were more susceptible to the three essential oils than the adults of their hosts C. maculatus. In the presence of cowpea seeds, the LC50s of the three essential oils were lower than in their absence, suggesting that the seeds may absorb a part of the volatiles. High doses of three essential oils slightly affected the survival of the fourth instar or the pupae of C. maculatus. This high survival was due to protection of larvae from volatiles by the surrounding seeds. The D. basalis were more affected by the oil volatiles than their hosts. Sub-lethal doses of essential oils reduced the duration of the adult life of both insect species and fecundity of the females. The differences in sensitivity of the host and its parasitoid could influence their population dynamics. The introduction of the essential oils into storage systems potentially could reduce density of parasitoid populations and increase seed losses. PMID- 11942756 TI - Integration of nonchemical treatments for control of postharvest pyralid moths (Lepidoptera: Pyralidae) in almonds and raisins. AB - We propose a treatment strategy combining an initial disinfestation treatment with one of three protective treatments as an alternative for chemical fumigation of almonds and raisins for control of postharvest insect populations. Initial disinfestation treatments using low oxygen controlled atmosphere (0.4% O2) were designed to disinfest product of field populations of pyralid moths; navel orangeworm, Amyelois transitella (Walker), in almonds and raisin moth, Cadra figulilella (Gregson), in raisins. The protective treatments were cold storage (10 degrees C), controlled atmosphere (5% O2) storage, and application of the Indianmeal moth granulosis virus, and were designed to prevent establishment of Indianmeal moth, Plodia interpunctella (Hubner). The initial disinfestation treatment was effective against laboratory populations of navel orangeworm and raisin moth. Efficacy of protective treatments was determined by exposure of commodities to laboratory Indianmeal moth populations at levels far higher than those found in commercial storage facilities. All three protective treatments prevented development of damaging Indianmeal moth populations as measured by pheromone trap catches and evaluation of product samples. Quality analysis by commercial laboratories showed that overall product quality for all protective treatments was maintained at levels acceptable by industry standards. PMID- 11942755 TI - Bioactivities of the leaf essential oil of Curcuma longa (var. ch-66) on three species of stored-product beetles (Coleoptera). AB - Essential oil extracted from the leaves of turmeric, Curcuma longa L., was investigated for contact and fumigant toxicity and its effect on progeny production in three stored-product beetles, Rhyzopertha dominica F. (lesser grain borer), Sitophilus oryzae L. (rice weevil), and Tribolium castaneum Herbst (red flour beetle). Oviposition-deterrent and ovicidal actions of C. longa leaf oil were also evaluated against T. castaneum. The oil was insecticidal in both contact and fumigant toxicity assays. The adults of R. dominica were highly susceptible to contact action of C. longa leaf oil, with LD50 value of 36.71 microg/mg weight of insect, whereas in the fumigant assay, adults of S. oryzae were highly susceptible with LC50 value of 11.36 mg/liter air. Further, in T. castaneum, the C. longa oil reduced oviposition and egg hatching by 72 and 80%, respectively at the concentration of 5.2 mg/cm2. At the concentration of 40.5 mg/g food, the oil totally suppressed progeny production of all the three test insects. Nutritional indices indicate >81% antifeedant action of the oil against R. dominica, S. oryzae and T castaneum at the highest concentration tested. PMID- 11942757 TI - Factors affecting capture of Cryptolestes ferrugineus (Coleoptera: Laemophloeidae) in traps placed in stored wheat. AB - Cryptolestes ferrugineus (Stephens), the rusty grain beetle, infests grain externally and is a common pest of stored wheat throughout the world. Detection and population estimation of this insect are important in avoiding discounts at the point of sale. Laboratory experiments compared number of insect captures in the WNB II probe and PC trap in stored grain with a known insect density. Capture rates were strongly related to insect densities in wheat. In a simultaneous test of insect density, ranging from one to three insects per kilogram, and temperatures between 20 and 40 degrees C, insect captures in WB II probe traps increased linearly with insect density in the grain but had a quadratic response to temperature. Hole density, ranging from 40 to 120 holes along a 15-cm stretch of the trap body, was unrelated to number of insect captures. Probe trap diameters ranging from 26 to 60 mm were also unrelated to insect captures. Finally, dead rusty grain beetles were recovered in probe traps. The recovery of dead insects increased with insect density when insects were found in an aggregated dispersal pattern, such as would be found following phosphine fumigation of grain. Experiments discussed here will help grain managers understand how probe traps may be used in C. ferrugineus population estimation. PMID- 11942758 TI - Effect of cold treatment on survival and development of codling moth (Lepidoptera: Tortricidae) in cherry. AB - 'Bing' cherries, Prunus avium L., were obtained from an organic orchard and a conventional commercial orchard. The two groups were examined separately in replicated tests infested with each fruit initially infested with a first-instar codling moth, Cydia pomonella (L.). To simulate commercial postharvest holding conditions, the treatments were 0 (control), 1, 2, 4, 7, 10, and 14 d cold storage at 3.3 degrees C. The fruits were examined three or more times to determine larval survival, life stage, fruit condition, and fungal disease. Survival of first instars was affected only by cold storage durations of > or = 7 d. When infested with codling moth larvae, both organic and conventionally grown cherries quickly deteriorated from fungal diseases. The rate of moth development was estimated from the surviving larvae and was significantly different between organic and conventionally grown cherries for all instars except the second. PMID- 11942759 TI - Use of Tenebrio molitor (Coleoptera: Tenebrionidae) to recycle organic wastes and as feed for broiler chickens. AB - Several dried waste materials from different origins were used as a substrate to grow Tenebrio molitor L. Nutrient/amino acid values differed depending on both larval size/weight and substrate. These larvae were experimentally used as a broiler feedstuff. Seven-day-old chicks of a commercially available strain with an average weight of 126 g were randomly distributed into nine six-broiler groups. Three levels of Tenebrio molitor larvae (0, 5, and 10% dry weight) were used in a 19% protein content sorghum-soybean meal basal diet, to evaluate feed intake, weight gain, and feed efficiency. Results after 15 d showed no significant differences among treatments. These data indicate that Tenebrio molitor has the potential to be used as protein source for raising broilers. PMID- 11942760 TI - Pollinating bees (Hymenoptera: Apiformes) of U.S. alfalfa compared for rates of pod and seed set. AB - Alfalfa (=lucerne) flowers require visiting bees to trip the sexual column, thereby providing pollination and subsequent pod and seed set. Previous studies have compared the pollination values of different bee species solely by the speed with which they handle flowers and the proportion of visited flowers tripped. In this greenhouse study, five species of bees, including the three commercially managed U.S alfalfa pollinators, are likewise compared for their floral tripping frequencies. These bee species are also compared for the pod set and mature seed that results from their single visits to virgin flowers. Regardless of the identity of the pollinating bee, tripped flowers had the same probabilities of pod set and seed set. Thus, differences in the single-visit pollination efficiencies of the various bee species are entirely attributable to the proportion of visited flowers that they trip. Females of the alkali bee, Nomia melanderi Cockerell, and the alfalfa leafcutting bee, Megachile rotundata F, tripped 81 and 78% of visited flowers, respectively. Males of these species are significantly less effective (61 and 51%, respectively), but still significantly superior to the honey bee, Apis mellifera L. (22% of visited flowers tripped). These relationships are supported by field data for tripping frequencies. One candidate pollinator, Osmia sanrafaelae Parker, shows promise (44% tripped), but not the congeneric O. aglaia Sandhouse (13% tripped). PMID- 11942761 TI - Effects of fluvalinate and coumaphos on queen honey bees (Hymenoptera: Apidae) in two commercial queen rearing operations. AB - We conducted research on the potential impacts of fluvalinate and coumaphos on honey bee, Apis mellifera L., queen viability and health. Queens were reared in colonies that had been treated with differing amounts of both fluvalinate and coumaphos. Pre- and posttreatment samples of both wax and bees were collected from all of the colonies and analyzed for total concentrations of fluvalinate and coumaphos. All queens were measured for queen weight, ovarial weight, and number of sperm in the spermathecae. The queens treated with high doses of fluvalinate weighed significantly less than low-dose or control queens, but otherwise appeared to develop normally. The highest fluvalinate concentrations were observed in the wax and queen cells of the high-dose group. The developing queens in colonies treated with as little as one coumaphos-impregnated strip for more than 24 h suffered a high mortality rate. Several of the queens showed sublethal effects from the coumaphos, including physical abnormalities and atypical behavior. The queens exposed to coumaphos weighed significantly less and had lower ovary weights than the control group queens. The highest coumaphos concentrations were observed in the queen cells and wax of the high-dose groups. PMID- 11942763 TI - A method for laboratory studies on the polyphagous predator Macrolophus pygmaeus (Hemiptera: Miridae). AB - A method for studying the life history parameters of Macrolophus pygmaeus Rambur was developed and described. Fecundity of the predator was studied by placing one female and one male in a plastic cylinder in which a stem piece was used as an ovipositional substrate. Smaller cylinders were used to study the percentage of egg hatch of M. pygmaeus. Results demonstrated that this method is suitable for studying the fecundity and egg eclosion of M. pygmaeus. Allowing for easy modification, the method offers suitable conditions for maintenance of both insect and plant material and can be inexpensively modified for different experiments and the parts are easily constructed. The proposed method could also be employed for similar studies on other predatory hemipterans such as mirids and anthocorids and very likely on other phytophagous insect species that lay their eggs in or on plant stems. PMID- 11942762 TI - Beauveria bassiana as a pathogen of the Mexican fruit fly (Diptera: Tephritidae) under laboratory conditions. AB - Bioassays were carried out under controlled conditions (27 +/- 2 degrees C, 80 +/ 5% RH, and a photoperiod of 12:12 [L:D] h) to evaluate the effect of eight strains of the entomopathogenic fungus Beauveria bassiana upon larvae, pupae, and adult females of the Mexican fruit fly, Anastrepha ludens (Loew). Mortality of the immature stages was low, 2-8% in larvae and 0% in pupae. However, very high levels of mortality were obtained for adult flies, with values of 100, 98, and 98% for the strains Bb16, Bb24, and Bb26, respectively. LC50 values for these three strains ranged from 3.12 x 10(6) to 9.07 x 10(6) conidia/ml. Lethal time 50 (LT50) was 2.8, 3.7, and 4.2 d for Bb16, Bb26, and Bb24 strains, respectively, with an average LT50 of 4.4 d across all strains. The fungal mycelium emerged through the soft parts of the exoskeleton, such as the wing bases, mouth, intersegmental regions of the legs, and membranous regions of the abdomen, coxae, and neck. Maximum percentage sporulation ranged from 66.4 to 74.7% for the three most virulent strains. PMID- 11942764 TI - Effect of temperature on parasitism and host-feeding of Trichogramma turkestanica (Hymenoptera: Trichogrammatidae) on Ephestia kuehniella (Lepidoptera: Pyralidae). AB - The egg parasitoid Trichogramma turkestanica Meyer is being evaluated as a biological control agent against the Mediterranean flour moth, Ephestia kuehniella Zeller, in flour mills. The longevity, parasitism and host-feeding of the parasitoid at four constant temperatures (15-30 degrees C) has been determined in the laboratory. The highest fecundity occurred at intermediate temperatures. The number of host eggs killed by host-feeding per female was highest at the two lower temperatures. A very conservative estimate of host feeding showed that it accounts for approximately half of the mortality of host eggs at 20 and 25 degrees C and thus could constitute a major mortality factor for the flour moth population. PMID- 11942765 TI - Use of transgenic Bacillus thuringiensis Berliner corn hybrids to determine the direct economic impact of the European corn borer (Lepidoptera: Crambidae) on field corn in eastern Canada. AB - Transgenic corn expressing Bacillus thuringiensis Berliner (Bt corn) (Maximizer and Yieldgard hybrids, Novartis Seeds), non-Bt isolines and high-performance (check) hybrids were evaluated for European corn borer, Ostrinia nubilalis (Hubner), damage and grain yield in commercial strip plots across Ontario in 1996 and 1997. Bt corn hybrids reduced stalk tunneling damage by 88-100%. In 1996, minimal damage was found in locations where only one generation of European corn borer occurred per year. Bt corn proved its greatest potential for reducing the number and length of cavities below the primary ear in locations where two generations of European corn borer were present. A yield response to using Bt hybrids only occurred when levels of tunneling damage exceeded 6 cm in length. European corn borer infestations resulted in a 6 and 2.4% reduction in yield for 1996 and 1997, respectively, when Bt hybrids were compared with their non-Bt isolines. A linear relationship was found between tunnel length per plant in centimeters (x) and yield protection (%) obtained from using Bt corn (y) (y = 1.02 + 0.005x, r2 = 0.7217). At a premium of $34.58 Canadian (CDN) perhectare for Bt corn seed, an infestation of at least 6 cm of corn borer tunneling per plant was required to break even at a market price for corn of $2.50 per bushel CDN. During the period of study, low infestations (0-2 cm) of European corn borer occurred at 25% of the locations assessed, moderate infestations (4-6 cm) occurred at 42% of the locations, and high infestations (>6 cm) occurred at 33% of the locations. At a corn price of $3.00 per bushel CDN and seed premiums of $34.58 per hectare CDN, 5 cm of tunneling was required for a return on investment in Bt seed, comprising only 55% of the growers in the study. With infestations of more than 6 cm of tunneling occurring only 33% of the time, a return on seed investment would be realized in only one of three growing seasons. At a seed premium of $24.70 per hectare CDN per year, at least $74 per hectare CDN in the year of infestation would be required to make up for the two years of no return. In this study, a $74 per hectare CDN return at a corn price of $9.26 per hectare CDN with >16 cm of tunneling damage would have occurred only 7.3% of the time. PMID- 11942766 TI - Population dynamics of apple maggot (Diptera: Tephritidae) in south central Pennsylvania. AB - The apple maggot, Rhagoletis pomonella (Walsh), was monitored with baited yellow panels and red spheres in commercial orchards, abandoned orchards, and unsprayed backyard apple trees in 1998 and 1999. Apple maggot adults were captured in all apple habitats, but the capture levels in the abandoned orchards and unsprayed backyard trees tended to be higher than in the commercial orchards. Peak capture occurred between mid-July and late August in both years. Emergence cages seeded with infested fruits were used to investigate bivoltinism, which was observed in both years. PMID- 11942767 TI - Life history and spatial distribution of Oriental beetle (Coleoptera: Scarabaeidae) in golf courses in Korea. AB - Larval and adult activity of the oriental beetle Exomala orientalis (Waterhouse), a pest of turfgrass in Korea, was investigated at four golf clubs in Pusan, Korea, from 1995 to 1999. Adult emergence was first observed on the greens in late May with peak activity occurring 2 wk later. During the day, E. orientalis adults were most active between 1800 and 2200 hours. First instars were found mostly in early July, second instars mostly in late July, and third instars from August to April. The density of larvae in fixed plots decreased steadily from the time of egg laying to pupation: 667/m3 on 26 July, 267/m3 on 29 August, and 122/m3 on 2 October 1997. All the observed E. orientalis completed one generation per year. Adult females were observed feeding on flowers of a late-blooming variety of Japanese chestnut (Castanea crenata Sieb & Zucc). E. orientalis larval densities were higher in greens with Japanese chestnut nearby, and where magpie, Pica pica sericea (Gould), feeding was observed. More E. orientalis adults emerged from the right, left, and back of greens than from the front or middle. The intensity of emergence was inversely proportional to the amount of golfer traffic on various parts of the green. Counting emergence holes may be a way that golf course superintendents can predict which greens and tees are most likely to be damaged from E. orientalis larvae without destructive sampling. PMID- 11942768 TI - Effect of transgenic plants expressing high levels of a tobacco anionic peroxidase on the toxicity of Anagrapha falcifera nucleopolyhedrovirus to Helicoverpa zea (Lepidoptera: Noctuidae). AB - Wild type and corresponding transgenic tomato (Lycopersicon esculentum Miller) and two tobacco (Nicotiana spp.) plants that express high levels of a tobacco anionic peroxidase were used to determine what type of interactions occurred between peroxidase altered plant chemistry and the baculovirus Anagrapha falcifera nucleopolyhedrovirus (AfMNPV) for control of neonate corn earworms, Helicoverpa zea (Boddie). Transgenic plants expressed approximately five to 400 times higher peroxidase activity than corresponding tissues of wild type plants. The H. zea larvae typically fed 1.5 times less on transgenic compared with wild type leaf disks. There was only one experiment (of three with tomato leaves) where the larvae that fed on transgenic leaves were less susceptible to the virus based on nonoverlapping 95% confidence intervals for LC50 values. When the exposure dose was corrected for reduced feeding on the transgenic leaf disks, the insecticidal activity of the virus was not significantly different for larvae fed on transgenic versus wild type plants. Eight other experiments (with tomato and two species of tobacco) indicated either no significant effect or enhanced susceptibility (when corrected for feeding rates) to the virus of larvae fed on the transgenic leaves. These results indicate enhanced insect resistance in plants expressing high levels of a specific anionic peroxidase may be compatible with applications of AfMNPV. Potential reasons for this compatibility are discussed. PMID- 11942769 TI - Effect of greenbugs (Homoptera: Aphididae) on yield loss of winter wheat. AB - The effect of greenbug, Schizaphis graminum (Rondani), feeding on the yield of four winter wheat cultivars commonly grown in Oklahoma was studied. Cultivars tested were 'Karl', a recent derivative 'Karl-92', and '2163', all greenbug susceptible cultivars; and 'TAM-110', a cultivar with resistance to biotype E greenbugs. The objectives were to determine the effect of different greenbug densities during fall and spring on yield of winter wheat, and to develop mathematical models to quantify the effect of greenbugs on yield loss. The intensity of greenbug infestations achieved in plots by artificial infestation varied among years and growing seasons within a year, but was generally sufficient to cause a reduction in yield. Among yield components, the number of heads per square meter and the number of seeds per head were frequently negatively correlated with the accumulated number of greenbug-days per tiller. Seed weight was rarely affected by greenbug infestation. A regression model estimated yield loss for greenbug-susceptible cultivars at 0.51 kg/ha loss of yield per greenbug-day in years with near normal precipitation, and a loss of 1.17 kg/ha under severe drought conditions. The susceptible winter wheat cultivars exhibited similar yield loss in relation to the intensity of greenbug infestation, as indicated by a common slope parameter in the regression model. Results suggest that the model is robust for predicting yield loss for susceptible cultivars. PMID- 11942770 TI - Comparison of adult corn rootworm (Coleoptera: Chrysomelidae) sampling methods. AB - Studies were conducted in Kansas corn and soybean fields during 1997 to compare various sampling methods, traps, and trap components for capturing three species of adult corn rootworms: western (Diabrotica virgifera virgifera Leconte), southern (D. undecimpunctata howardi Barber), and northern (D. barberi Smith & Lawrence). Lure constituents affected the species of beetle attracted to the trap. Traps with a lure containing 4-methoxycinnamaldehyde attracted more western corn rootworms, those with a lure containing eugenol were more attractive to northern corn rootworms, and those containing trans-cinnamaldehyde were most attractive to southern corn rootworms. Multigard sticky traps caught more beetles than did Pherocon AM sticky traps. In corn, a newly designed lure trap caught more beetles than did sticky traps on most occasions. Also, lure-baited sticky traps caught more beetles than did nonbaited sticky traps. Varying the color of the lure trap bottom did not affect the number caught. In soybeans, the new lure traps captured more beetles than did the nonbaited Multigard or Pherocon AM sticky traps. Results of this study suggest the new lure trap may provide a more accurate assessment of corn rootworm populations than traditional monitoring techniques and may be more esthetically pleasing to growers and consultants. PMID- 11942771 TI - Increased hypothalamic dopaminergic tone only in early parous women with either malignant or benign breast tumors. AB - OBJECTIVE: To investigate whether the serum prolactin (PRL) response to a dopamine antagonist was different in nonobese, euthyroid women with malignant or benign breast tumors in comparison with healthy women, considering their age at first full-term pregnancy or their nulliparity. METHODS: Serum PRL concentrations before and 60, 90, and 120 minutes after oral administration of metoclopramide (10 mg) were studied in 122 nonobese, nonsmoking, euthyroid women: 28 who had invasive breast cancer, stage I or II (group 1), 34 who had benign breast disease (group 2), and 60 who were clinically healthy (group 3). These three main groups were subdivided into early and late parous women (< or = 25 and >25 years, respectively) and nulliparous women, and the menopausal status was also considered. RESULTS: Early parous women with invasive breast cancer (both premenopausal and postmenopausal) and early parous premenopausal women with benign breast disease had significantly higher serum PRL concentrations in response to administration of metoclopramide (P<0.05) and a greater area under the PRL curve (P<0.05) than those observed in early parous healthy women. No other significant differences in the serum PRL response were noted between or within groups. CONCLUSION: The results of this study suggest the existence of an increased hypothalamic dopaminergic tone in early parous women but not in late parous or nulliparous women with malignant or benign breast tumors in comparison with similar healthy women. This finding may represent an adaptive protective mechanism attempting to prevent persistently increased serum PRL concentrations, a factor that could adversely affect the clinical evolution of the disease. PMID- 11942772 TI - Fenofibrate of gemfibrozil for treatment of types IIa and IIb primary hyperlipoproteinemia: a randomized, double-blind, crossover study. AB - OBJECTIVE: To compare the hypolipidemic effects of gemfibrozil and micronized fenofibrate in patients with primary hyperlipoproteinemia, phenotypes IIa and IIb, with emphasis on their cholesterol-lowering effectiveness. METHODS: A randomized, double-blind, double-dummy, crossover study was performed to assess the effects of gemfibrozil (900 mg) and micronized fenofibrate (200 mg), administered once daily, to 21 patients (45 to 70 years old)-16 with type IIa and 5 with type IIb primary hyperlipidemia. The two treatment periods lasted 6 weeks each; the run-in and washout periods were 4 weeks. RESULTS: Both drugs significantly reduced total cholesterol, calculated low-density lipoprotein (LDL) cholesterol, triglycerides, apolipoprotein B, and fibrinogen (P<0.01 for all calculations, except P<0.05 for fibrinogen with gemfibrozil therapy) and increased high-density lipoprotein (HDL) cholesterol (P<0.01). Neither drug affected Lp(a) lipoprotein, whereas uric acid was reduced only by fenofibrate (P<0.01). The percentage decrease in total cholesterol and LDL cholesterol was greater with fenofibrate than with gemfibrozil (-22% versus -15%, P<0.02; and 27% versus -16%, P<0.02, respectively). In contrast, reductions in levels of triglycerides (-54% versus -46.5%), apolipoprotein B, and fibrinogen, as well as the increase in HDL (+9% for both drugs), showed no significant difference between treatments. Separate analysis of patients with type IIb hyperlipoproteinemia showed essentially the same plasma lipid changes as for the overall group, but with greater modifications in triglyceride and HDL concentrations. CONCLUSION: Fenofibrate and gemfibrozil induced similar variations from baseline values in triglycerides, HDL cholesterol, apolipoprotein B, and fibrinogen, but the decreases in total and LDL cholesterol levels were greater with fenofibrate, in this group of patients with primary hyperlipidemia. PMID- 11942774 TI - Parental perception and factors associated with glycemic control in pediatric patients with type 1 diabetes. AB - OBJECTIVE: To compare subjective parental perceptions of short-term diabetes control with objective glycated hemoglobin (HbA1c) measurements in a pediatric population. METHODS: Parents of 159 children and adolescents with type 1 diabetes anonymously completed a questionnaire concerning demographics, parental perception of the child's recent glycemic control, concern about nocturnal blood glucose levels, and factors related to severity of the child's diabetes. Nonparametric correlations were performed among all variables and the child's current HbA1c measurement. Good short-term glycemic control was defined as an HbA1c value of < or = 8.5%. RESULTS: Five factors were found to correlate significantly with good glycemic control, and these variables were subjected to a logistic regression analysis. With use of a cutoff value of 0.58, results explained 26% of the variance and had an overall 71% correct classification rate. Children with good glycemic control were younger, had a shorter duration of diabetes, lived in families with higher socioeconomic status, were less likely to experience hospitalizations due to their diabetes, and had parents who "always worried" about nocturnal blood glucose variations. Parental perception and HbA1c values were not significantly related (P>0.19); 60% of the parents correctly perceived the status of diabetes in their children, and 40% misperceived the level of glycemic control. Further analysis suggested that accuracy of parental perception was highly dependent on the child's level of glycemic control. Parents whose children had lower HbA1c values (< or = 7.5%) correctly perceived the status of blood glucose control in their children, whereas parents whose children had poorer glycemic control (HbA1c > or = 9.5%) had inaccurate perceptions. CONCLUSION: Subjective parental perceptions of diabetes control in their children may not correspond to objective assessments. Our findings suggest a need for more educational feedback concerning HbA1c levels and recommended goals for glycemic control. PMID- 11942773 TI - Effect of thiazolidinediones on high-density lipoprotein subfractions. AB - OBJECTIVE: To evaluate the effects of thiazolidinediones (TZDs) on specific high density lipoprotein (HDL) subfractions (HDL2 and HDL3) in patients with type 2 diabetes. METHODS: We conducted a case-control study in a university diabetes clinic. The study included 45 patients with type 2 diabetes mellitus and dyslipidemia; 31 of these patients received a TZD (troglitazone), and 14 patients received metformin. At the time of the study, the mean duration of therapy was 7 and 11 months, respectively. RESULTS: In the TZD group, a significant increase of 12.6% occurred in the total HDL levels (from 34.9 +/- 2.2 mg/dL to 39.3 +/- 2.2 mg/dL; P = 0.0008), whereas no significant change was noted in the metformin group (from 40.9 +/- 3.5 mg/dL to 40.1 +/- 3.2 mg/dL; P = 0.14). All the increase in total HDL levels in the TZD group was due to an increase in the HDL3 subfraction, which rose by 16.7% (from 26.9 +/- 0.9 mg/dL to 31.4 +/- 1.2 mg/dL; P = 0.008). The HDL2 subfraction remained essentially unchanged (from 8.1 +/- 1.7 mg/dL to 7.9 +/- 1.4 mg/dL; P = 0.80). In the metformin group, no significant changes were found in either the HDL3 or the HDL2 subfractions. CONCLUSION: The TZD-induced increase in total HDL levels in our patients was entirely attributable to an increase in the HDL3 subfraction. PMID- 11942775 TI - Insulinoma in a patient with tuberous sclerosis: is there an association? AB - OBJECTIVE: To describe a patient with tuberous sclerosis who, on initial assessment, had neurologic symptoms, which were ultimately found to be caused by an insulinoma. METHODS: We present a case report with clinical, laboratory, and radiologic data. The literature is reviewed relative to tuberous sclerosis and islet cell tumors, and a possible association is discussed. RESULTS: A 43-year old man with a history of tuberous sclerosis required medical attention because of mental confusion and slurred speech and was found to have hypoglycemia. Neuroradiologic imaging showed no new lesions to account for his symptoms. His physical examination was striking for a large abdominal mass, which showed increased uptake on octreotide scanning. After surgical resection, the mass measured 21 cm and was found to be an insulinoma. Blood glucose values were normal postoperatively and on follow-up, and the patient had no recurrence of the symptoms. CONCLUSION: From this report, we emphasize two findings. First, we draw clinicians' attention to the possibility of an association between islet cell tumors and tuberous sclerosis and suggest consideration of this diagnosis in patients with tuberous sclerosis who have new or worsening neurologic symptoms. Second, the insulinoma we describe is, to our knowledge, the largest to be reported thus far in the literature. PMID- 11942776 TI - Diagnosis of acromegaly in a patient with type 1 diabetes mellitus. AB - OBJECTIVE: To describe a case of acromegaly in a young patient with poorly controlled diabetes mellitus and suggest guidelines to distinguish acromegaly and high growth hormone (GH) levels previously reported in poorly controlled type 1 diabetes. METHODS: We present a detailed case report, including clinical findings and serial laboratory results in a patient with type 1 diabetes and a GH secreting pituitary tumor. RESULTS: A 28-year-old woman with type 1 diabetes underwent assessment for secondary amenorrhea and worsening glycemic control. A low estradiol level and an inappropriately low level of follicle-stimulating hormone prompted magnetic resonance imaging of the head, which demonstrated a pituitary adenoma. Subsequent endocrine investigation revealed a high insulin like growth factor I (IGF-I) level (849 mg/L; normal range, 122 to 400). The concentration of insulin-like growth factor-binding protein-3 (IGFBP-3) was also elevated (5.5 mg/L; normal range, 2.0 to 4.2). GH levels measured during episodes of spontaneous hyperglycemia (>180 mg/dL) were in the range of 3 to 5 ng/mL and failed to suppress to below 2 ng/mL after a bromocriptine suppression test. The patient underwent transsphenoidal resection of a pituitary tumor, which stained positively for GH by immunohistochemistry. Postoperatively, glycemic control improved, with decreased fluctuations of hypoglycemia and hyperglycemia, despite a decrease in insulin requirements. CONCLUSION: This report highlights the difficulty in interpreting GH and IGF-I levels in patients with type 1 diabetes. In addition, a detailed review of the literature suggests that IGFBP-3 measurements may be helpful in confirming the diagnosis of concurrent acromegaly and distinguishing it from high GH levels attributable to poor control of diabetes. PMID- 11942777 TI - Cushing's syndrome manifesting as pseudo-central hypothyroidism and hyperosmolar diabetic coma. AB - OBJECTIVE: To report an unusual case of Cushing's syndrome caused by an adrenal pheochromocytoma, manifesting as pseudo-central hypothyroidism and diabetic hyperosmolar coma. METHODS: A detailed case report is presented, including clinical, laboratory, and radiologic findings as well as results of selective adrenal vein sampling. RESULTS: In a 69-year-old woman with weight gain and hypothyroidism, diabetes mellitus with hyperosmolar coma developed precipitously. She had mild hypertension, but no diabetes was noted 2 weeks before the hyperosmolar event. Evaluation revealed Cushing's syndrome due to ectopic secretion of adrenocorticotropic hormone from an adrenal pheochromocytoma. After surgical resection of the tumor, the diabetes and the hypertension resolved. Furthermore, the pseudo-central hypothyroidism was eliminated, but primary hypothyroidism was unmasked. This combination has not been reported previously. CONCLUSION: This case illustrates the array of endocrinopathies that can be associated with pheochromocytoma, causing Cushing's syndrome. PMID- 11942779 TI - What's in the "sellar"? PMID- 11942778 TI - Obesity in transplant patients: case report showing interference of orlistat with absorption of cyclosporine and review of literature. AB - OBJECTIVE: To report a case of an obese patient who had undergone renal transplantation and who had subtherapeutic levels of serum cyclosporine after treatment with orlistat. METHODS: The clinical and laboratory findings are presented, and the few cases reported in the literature are reviewed. RESULTS: A 29-year-old woman had subtherapeutic plasma levels of cyclosporine after orlistat treatment (360 mg/day) was initiated. The subtherapeutic levels persisted even though orlistat was administered the recommended 2 hours before ingestion of cyclosporine and even though the dosage of orlistat was decreased to only 240 mg/day. Because an increase of body weight is common after organ transplantation, treatment with orlistat has been used. In such patients, however, six cases of reduced therapeutic plasma levels of cyclosporine have been reported. Although a drug-drug interaction has been suggested, this case suggests that the decreased plasma cyclosporine levels are due to reduced absorption of fats rather than a drug-drug interaction. Because this patient was unable to adhere to a low-fat diet, she experienced severe diarrhea, a factor that may have dramatically diminished the absorption of cyclosporine. CONCLUSION: Adherence to a low-fat diet should be strongly recommended if orlistat is prescribed to patients taking cyclosporine. Moreover, strict surveillance of the plasma concentration of cyclosporine is important. PMID- 11942780 TI - Introduction: nitric oxide chemistry. PMID- 11942781 TI - Photoinduced linkage isomers of transition-metal nitrosyl compounds and related complexes. PMID- 11942782 TI - Infrared spectra and density functional theory calculations on transition metal nitrosyls. Vibrational frequencies of unsaturated transition metal nitrosyls. PMID- 11942783 TI - Nitrogen NMR spectroscopy of metal nitrosyls and related compounds. PMID- 11942784 TI - Coordination and organometallic chemistry of metal-NO complexes. PMID- 11942785 TI - Mechanistic aspects of the reactions of nitric oxide with transition-metal complexes. PMID- 11942786 TI - Interactions of organic nitroso compounds with metals. PMID- 11942787 TI - Solid-state structures of metalloporphyrin NO(x )compounds. PMID- 11942788 TI - Nitric oxide donors: chemical activities and biological applications. PMID- 11942789 TI - Chemistry of the nitric oxide-releasing diazeniumdiolate ("nitrosohydroxylamine") functional group and its oxygen-substituted derivatives. PMID- 11942790 TI - Non-heme iron nitrosyls in biology. PMID- 11942791 TI - Nitric oxide and myoglobins. PMID- 11942792 TI - Intrinsic and extrinsic modulation of nitric oxide synthase activity. PMID- 11942793 TI - Mechanism of free-radical generation by nitric oxide synthase. PMID- 11942794 TI - Nitric oxide in biological denitrification: Fe/Cu metalloenzyme and metal complex NO(x) redox chemistry. PMID- 11942796 TI - A new mechanism for metal-catalyzed stannane dehydrocoupling based on alpha-H elimination in a hafnium hydrostannyl complex. AB - The hafnium hydrostannyl complex CpCpHf(SnHMes(2))Cl (Cp = eta(5)-C(5)Me(5)) has been synthesized from the reaction of CpCpHf(H)Cl with Mes(2)SnH(2). This compound has been identified as an intermediate in the metal-catalyzed dehydrocoupling of Mes(2)SnH(2) to the distannane Mes(2)HSnSnHMes(2) (Mes = 2,4,6 trimethylphenyl). The dehydrocoupling in this system appears to occur by elimination of :SnMes(2) from CpCpHf(SnHMes(2))Cl, with Sn-Sn bond formation proceeding via insertion of the stannylene into a Sn-H bond. PMID- 11942795 TI - Bioimaging of nitric oxide. PMID- 11942797 TI - Absolute configuration of D(2)-symmetric fullerene C(84). AB - The D(2)-symmetric isomer of fullerene C(84) predominantly found in soots is inherently chiral. We determine its absolute configuration by comparison of the experimental electronic circular dichroism (CD) spectrum with time-dependent density functional calculations. PMID- 11942798 TI - CO-transfer carbonylation reactions. A catalytic Pauson-Khand-type reaction of enynes with aldehydes as a source of carbon monoxide. AB - The reaction of enynes with aldehydes in the presence of a catalytic amount of [RhCl(cod)](2)/dppp results in the Pauson-Khand-type reaction without the use of gaseous carbon monoxide to give bicyclic cyclopentenones in high yields (14 examples). Aldehydes serve as a source of carbon monoxide, and their carbonyl moiety is transferred to enynes, resulting in the formation of the carbonylated products. This reaction represents the first example of a CO-transfer carbonylation. PMID- 11942799 TI - Asymmetric Diels-Alder reactions catalyzed by a triflic acid activated chiral oxazaborolidine. AB - This paper reports a new method for the generation of chiral Lewis superacids by protonation of a non-Lewis acidic oxazaborolidine (1) with triflic acid. The resulting cationic species (3) are powerful and highly enantioselective catalysts for the Diels-Alder reaction of various 1,3-dienes with alpha,beta-enals. An optimization study (see Table 1) led to the selection of reaction conditions and catalysts (6A and 6B) which are very effective. The reactions are simple to conduct, reproducible, and economical, since only ca. 6 mol % of catalyst is required. In addition, the chiral catalyst precursor is readily recovered for reuse (>95% efficiency) and is commercially available. The broad scope of the process is documented by the 14 examples listed in Table 2. The absolute stereochemical course of the Diels-Alder reactions catalyzed by 6A and 6B was successfully predicted on the basis of the mechanistic principles which have recently been formulated for this type of catalytic enantioselective reaction involving re-face attack by the diene on complex 7. The mode of generation of Lewis superacids 6A and 6B allows an approximate comparison (or scale) connecting the catalytic power Lewis and protic acids. PMID- 11942800 TI - Nonpeptide inhibitors of cathepsin G: optimization of a novel beta-ketophosphonic acid lead by structure-based drug design. AB - The serine protease cathepsin G (EC 3.4.21.20; Cat G), which is stored in the azurophilic granules of neutrophils (polymorphonuclear leukocytes) and released on degranulation, has been implicated in various pathological conditions associated with inflammation. By employing high-throughput screening, we identified beta-ketophosphonic acid 1 as a moderate inhibitor of Cat G (IC(50) = 4.1 microM). We were fortunate to obtain a cocrystal of 1 with Cat G and solve its structure by X-ray crystallography (3.5 A). Structural details from the X-ray analysis of 1.Cat G served as a platform for optimization of this lead compound by structure-based drug design. With the aid of molecular modeling, substituents were attached to the 3-position of the 2-naphthyl ring of 1, which occupies the S1 pocket of Cat G, to provide an extension into the hydrophobic S3 region. Thus, we arrived at analogue 7 with an 80-fold potency improvement over 1 (IC(50) = 53 nM). From these results, it is evident that the beta-ketophosphonic acid unit can form the basis for a novel class of serine protease inhibitors. PMID- 11942801 TI - Cascade radical reactions catalyzed by a Pd/light system: cyclizative multiple carbonylation of 4-alkenyl iodides. AB - Cascade reactions of 4-alkenyl iodides, involving a carbonylation-cyclization carbonylation sequence, were accomplished by a hnu/Pd system. The stereochemical outcomes suggest that radical carbonylation and subsequent acyl radical cyclization may be involved in this reaction. PMID- 11942803 TI - Rapid cross-disproportionation between superoxometal ions and acylperoxyl radicals. AB - A superoxochromium complex Cr(aq)OO(2+) reacts with acetylperoxyl radicals, CH(3)C(O)OO(*), with a rate constant of 1.49 x 10(8) M(-1) s(-1). The kinetics were determined by laser flash photolysis, using an organocobalt complex as a radical precursor and ABTS(*-) as a kinetic probe. The initial step is believed to involve radical coupling at the remote oxygen of Cr(aq)OO(2+), followed by elimination of O(2) and formation of CH(3)COOH and Cr(V)(aq)O(3+). The latter disproportionates and ultimately yields Cr(aq)(3+) and HCrO(4)(-). PMID- 11942802 TI - Decay of the transient Cu(B)-CO complex is accompanied by formation of the heme Fe-CO complex of cytochrome cbb(3)-CO at ambient temperature: evidence from time resolved Fourier transform infrared spectroscopy. AB - Time-resolved step-scan Fourier infrared spectroscopy has been used to study the CO-bound cbb(3)-type cytochrome c oxidase from Pseudomonas stutzeri at room temperature. We observe a single band in the FTIR spectrum at 1956 cm(-1) (beta form). The time-resolved data indicate that upon photolysis, CO is transferred from heme b(3) (nu(CO) = 1956 cm(-1)) to CuB (nu(CO) = 2064 cm(-1)). The decay of the 2065 cm(-1) peak (t(1/2) = 120 +/- 16 ms) and the development of the 1956 cm( 1) peak (t(1/2) = 144 +/- 8 ms ) suggest that formation of the Fe-CO complex is concurrent with the decay of the CuB-CO complex. The intensity ratio of the Fe CO/CuB-CO (2.15) remains constant for all data points, and thus we conclude that no fraction of CO escapes the binuclear center at 293 K. PMID- 11942804 TI - Stable formally zerovalent and diamagnetic monovalent niobium and tantalum complexes based on diazadiene ligands. AB - This paper describes the efficient synthesis and full characterization of rare formally zerovalent and diamagnetic monovalent pseudooctahedral niobium and tantalum complexes. The reaction of NbCl(4)(thf)(2) with 4 equiv of Na and 3.5 equiv of iPr(2)-dad (1,4-diisopropyl-1,4-diaza-1,3-diene) yields Nb(iPr(2) dad)(3) in 52% yield. Ta(iPr(2)-dad)(3) is also obtained in 33% yield from 5 equiv of Na/naphthalene and 3.5 equiv of iPr(2)-dad. Oxidation of these complexes with AgBPh(4) yields the diamagnetic complexes [M(iPr(2)-dad)(3)][BPh(4)] (M = Nb, Ta). X-ray and spectroscopic data indicate that the unpaired electron density is localized on the ligands. DFT calculations reveal that, in the prevailing D(3) symmetry, a very strong splitting of t(2g) metal-based orbitals occurs, leading to the diamagnetism of the 16e M(iPr(2)-dad)(3)(+). This strong splitting allows a M-N nonbonding, ligand-based orbital to accommodate additional electrons, as a result of which the formally zerovalent complexes, M(iPr(2)-dad)(3), are in fact correctly formulated as M(+)iPr(2)-dad (-), that is, (16e + 1e). PMID- 11942805 TI - Bio-barcodes based on oligonucleotide-modified nanoparticles. AB - By utilizing oligonucleotide-modified Au nanoparticles encoded with sequences that act as biobarcodes, one can screen for multiple target polyvalent proteins simultaneously in one solution. This novel concept was demonstrated with two types of detection formats, a homogeneous assay and one based on oligonucleotide microarrays. With such an approach, one can prepare an extraordinarily large number of barcodes from synthetically accessible oligonucleotides (e.g., a 12-mer sequence offers 4(12) possible barcodes). PMID- 11942807 TI - Osmium tetroxide-promoted catalytic oxidative cleavage of olefins: an organometallic ozonolysis. AB - A mild, organometallic alternative to ozonolysis utilizing oxone and OsO(4) is presented. This is a direct oxidation of olefins via the carbon-carbon cleavage of an osmate ester by the action of oxone. Twenty-four different olefins were converted to their corresponding ketones or carboxylic acids in high yields (>80%). Free alcohols, acetate- and benzyl-protected alcohols, and 1,2-diols were stable under these conditions. This method should be applicable for traditional organic synthesis. PMID- 11942806 TI - Anionic cyclization of a cross-conjugated enediyne. AB - Cross-conjugated enediynes cannot follow the Bergman cycloaromatization as it involves a methylenediyne moiety with only five pi e(-), insufficient for aromatization. Under reductive conditions the cyclization is made feasible by generating a product with a Huckel number of pi electrons. We illustrate this principle and demonstrate for the first time an anionic cyclization of a cross conjugated enediyne that results in formation of a five-membered ring. 9-(3 Phenyl-1-phenylethynylprop-2-ynylidene)-9H-fluorene (3) was reduced by potassium to yield the dianion of 9-(3,4-diphenylcyclopenta-2,4-dienylidene)-9H-fluorene (4(2-)), which contains a cyclopentadienyl fragment, and oxidation with iodine yielded the unstable fulvalene 4. PMID- 11942808 TI - Using "internal free volume" to increase chromophore alignment. AB - Triptycenes have general applicability for increasing the alignment of fluorescent and dichroic dyes in LC hosts. Dyes containing varying numbers of triptycenes were synthesized to study the effect of free-volume alignment of triptycenes on the alignment of dyes. These dyes were designed such that multiple triptycenes could be incorporated and the triptycene-free volume is coincident to the aspect ratio of the dye, allowing a cooperative effect to increase their overall average alignment. With increasing triptycene incorporation, a stepwise increase in the alignment parameters of each dye was seen. It was also found that the attachment of one triptycene group has a negligible effect on the optical switching response times of the dyes. This can be a powerful tool for designing dyes with higher alignments for a variety of applications including guest-host reflective LCDs and holographic data storage. PMID- 11942809 TI - A full interionic potential for Na(1+x)Zr(2)Si(x)P(3-x)O(12) superionic conductors. AB - Most inorganic solids are made up of octahedral and tetrahedral units interconnected to give an infinite framework. Use of computer simulation to study these materials has not been as prevalent as in the organic or biomolecules. Na(1+x)Zr(2)Si(x)P(3-x)O(12) is a typical inorganic solid with ZrO(6) octahedra and (Si/P)O(4) tetrahedra which are shown along with a few Na(+) sites marked M1, M2, and M3. We report here a full interionic potential which reproduces the structure and conductivity of these solids. This augurs well for the study of other inorganic solids. PMID- 11942810 TI - Excited-state reactions of an isolable silylene with aromatic compounds. AB - The first intermolecular reactions of the excited state of a silicon divalent compound (silylene) with benzene derivatives were discovered. Typically, when a benzene solution of an isolable silylene is irradiated with light of wavelengths longer than 420 nm at room temperature, the corresponding silacyclohepta-2,4,6 triene (silepin) is yielded quantitatively. The photochemical insertion of the silylene toward substituted benzenes occurs in general to give the corresponding substituted silepins. The insertion reaction is highly sensitive to the steric hindrance at a reacting C-C double bond in benzene; during the reactions of the silylene with substituted benzenes, only unsubstituted C-C double bonds in the benzene ring reacted selectively. The irradiation of the silylene in the presence of mesitylene afforded the insertion product to a benzylic C-H bond, indicative of the biradical nature of the excited-state silylene. PMID- 11942811 TI - Hyperfine fields at the Li site in LiFePO(4)-type olivine materials for lithium rechargeable batteries: a (7)Li MAS NMR and SQUID study. AB - The (7)Li NMR isotropic shift for olivine LiMPO(4) (M = Fe, Mn, Co, Ni) is assigned to hyperfine coupling between the (7)Li nucleus and the transition metal unpaired electrons on the basis of the Curie-Weiss temperature dependence of the shift. The hyperfine shift arises from a linear combination of Li-O-M through bond interactions wherein the unpaired A' electrons contribute a negative shift and the unpaired A' ' electrons contribute a positive shift. The hyperfine coupling constant is determined for each composition. PMID- 11942812 TI - Positive microcontact printing. AB - Microcontact printing alkanethiols from an inked, microstructured stamp onto Au or Cu results in the formation of a self-assembled monolayer, which can locally protect the substrate from wet etching. This lithographic technique resembles a negative-type of lithography but can be inverted to the positive process. This is done by printing a type of oligothiol that does not protect the substrate from etching but prevents the adsorption of a protective monolayer from solution. PMID- 11942813 TI - Hydrothermal assembly of a novel three-dimensional framework formed by [GdMo(12)O(42)](9-) anions and nine coordinated Gd(III) cations. AB - The three-dimensional framework of [Gd(H(2)O)(3)](3)[GdMo(12)O(42)]vdt.3H(2)O, which was synthesized from hydrothermal reaction, is built up from Silverton-type anions linked by nine coordinated gadolinium(III) cations. It is the first time that the paramagnetic lanthanide(III) cation is introduced into the Silverton type anionic center in the heteropolymolybdate chemistry, and it acts as 18 dentate ligands coordinating to six Gd(III) atoms and linking up six other neighboring [Mo(12)GdO(42)](9-) units in a staggered manner to generate a remarkable three-dimensional framework. The magnetic susceptibility measurement suggests that the weak antiferromagnetic behavior results from the transformation of O-Mo-O units, which has been confirmed by EPR spectra. PMID- 11942814 TI - Tetraphenyl-p-benziporphyrin: a carbaporphyrinoid with two linked carbon atoms in the coordination core. AB - Replacement of one of the pyrrole rings with a p-phenylene unit transforms porphyrin into p-benziporphyrin (1), an aromatic carbaporphyrinoid that locates two connected carbon atoms in the coordination core. p-Benziporphyrin forms a complex with cadmium(II) (2) with an unprecedented eta(2) Cd(II)-arene interaction. PMID- 11942815 TI - Design and synthesis of a fluorescent reporter of protein kinase activity. AB - There is widespread interest in developing fluorescent reporters of protein kinase activity, species that can furnish a visual readout of both when and where intracellular kinases are activated in response to a stimulus. We have constructed and identified, via a combination of rational design, library synthesis, and screening, a difluorofluorescein-appended peptide-based species that responds to protein kinase C phosphorylation in a fluorescently sensitive fashion. The phosphorylation-induced divalent metal ion-mediated 265% enhancement in fluorescence proceeds with a V(max) of 8.5 micromol/min.mg and a K(m) of 20.5 microM. PMID- 11942816 TI - Retention of configuration on the oxidative addition of P-H bond to platinum (0) complexes: the first straightforward synthesis of enantiomerically pure P-chiral alkenylphosphinates via palladium-catalyzed stereospecific hydrophosphinylation of alkynes. AB - The oxidative addition of pure (R(P))-menthyl phenylphosphinate 1 to a platinum (0) complex proceeds readily with retention of configuration at the chiral phosphorus center which was unambiguously confirmed by an X-ray analysis. In the presence of a catalytic amount of palladium, the hydrophosphinylation of a variety of alkynes with 1 also takes place stereospecifically, with retention of configuration, affording high yields of the corresponding vinylphosphinates bearing a single chirality at phosphorus. PMID- 11942817 TI - An oft-studied reaction that may never have been: direct catalytic conversion of methanol or dimethyl ether to hydrocarbons on the solid acids HZSM-5 or HSAPO-34. AB - Using highly purified reagents and careful tests, we show that methanol and dimethyl ether are apparently unreactive on the two most important methanol-to hydrocarbon catalysts, HZSM-5 and HSAPO-34. Thus, none of the "direct" mechanisms involving two to four carbon atoms in intermediates such as oxonium ylides, carbenes, carbocations, and free radicals are applicable. Only the "indirect" route (hydrocarbon pool) is an established mechanism for this chemistry. An active catalyst requires a hydrocarbon pool that typically begins with products from organic impurities in the feed, carrier gas, or the solid acid itself. Impurities may also play important roles in other reactions catalyzed by solid acids. PMID- 11942818 TI - Monomeric phosphido and phosphinidene complexes of nickel. AB - An unusual family of three-coordinate, d(8) and d(9) nickel phosphido and phosphinidene complexes containing the chelating 1,2-bis(di-tert butylphosphino)ethane (dtbpe) ligand and a terminal PR(2)(-) or PR(2-) ligand have been prepared. The complexes (dtbpe)Ni[P(t-Bu)(2)] (2), [(dtbpe)Ni[=P(t Bu)(2)](+)][PF(6)(-)] (3), [(dtbpe)Ni[=P(H)(dmp)](+)][PF(6)(-)] (5), and (dtbpe)Ni[=P(dmp)] (6) have been structurally characterized by single-crystal X ray diffraction methods. The three-coordinate d(8) complexes exhibit Ni-P bond lengths and ligand geometries that indicate they participate in symmetry-allowed ligand-to-metal pi bonding involving phosphorus p-electrons and a metal-orbital of pi symmetry that lies in the Ni coordination plane. Compound 6 is a rare example of a late-transition-metal terminal phosphinidene complex. PMID- 11942819 TI - An unusual palladium complex involved in an unusual rearrangement of ortho palladated aryldithioacetals. AB - The reaction of 1 with Pd(dba)(2), Tl(TfO) and PPh(3) in 1:1:1:2 molar ratios to give 3 implicates (i) an oxidative addition reaction, (ii) a rearrangement involving the cleavage of one HC-S bond and formation of an aryl-S bonds, and (iii) coordination of the Pd(PR(3))(2) group with a ligand intermediate between eta(2)-[ArCH=S((+))To] and kappa(2)-C,S-ArCH((-))STo, which requires the consideration of 3 as intermediate between a Pd(0) and a Pd(II) complex. The coordination of the ligand as a chelating three-membered ring, instead of the expected five-membered ring involving C(10) and S(1), and the partial intramolecular redox process are explained as a consequence of the transphobia of the pair of ligands Ph(3)P/CH(STo)Ar. PMID- 11942820 TI - Detection of a proton-transfer process by kinetic solvent isotope effects in NH(4)(+)-mediated reactions catalyzed by a hammerhead ribozyme. AB - Hammerhead ribozymes have been considered to be metalloenzymes. However, this proposal was recently questioned by the finding that the reaction proceeds in the presence of high concentrations of monovalent ions such as NH(4)(+) ions and in the absence of any divalent metal ions. Our present analysis based on solvent isotope effects indicates that (1) a proton transfer(s) occurs only in the NH(4)(+)-mediated reaction but not in metal-ion-mediated reactions such as Mg(2+) and Li(+)-mediated reactions, (2) the catalyst that stabilizes the 5' leaving group in the NH(4)(+)-mediated reaction is different from that in the metal-ion mediated HH ribozyme reactions, (3) an NH(4)(+) ion seems to act as a general acid catalyst, and (4) a nucleobase alone should not be the catalyst. PMID- 11942821 TI - Antihydrophobic solvent effects: an experimental probe for the hydrophobic contribution to enzyme-inhibitor binding. AB - The hydrophobic component to the binding affinities of one acyclic phosphinate (4) and three macrocyclic phosphonamidate inhibitors (1-3) to the zinc peptidase thermolysin was probed by varying the solvent composition. Increasing the percentage of ethanol in the buffer solution over the range 0-9% increases the inhibition constants, K(i), by up to an order of magnitude. This approach represents an experimental method for distinguishing solvation from conformational or other effects on protein-ligand binding. The size of the "antihydrophobic effect" is correlated with the amount of hydrophobic surface area sequestered from solvent on association of the inhibitor and enzyme, although it is attenuated from that calculated from the surface tension of ethanol-water mixtures. The results are consistent with the Lum-Chandler-Weeks explanation for the size dependence of the hydrophobic effect. PMID- 11942822 TI - Boundary conditions for the Swain-Schaad relationship as a criterion for hydrogen tunneling. AB - Hydrogen quantum mechanical tunneling has been suggested to play a role in a wide variety of hydrogen-transfer reactions in chemistry and enzymology. An important experimental criterion for tunneling is based on the breakdown of the semiclassical prediction for the relationship among the rates of the three isotopes of hydrogen (hydrogen -H, deuterium -D, and tritium -T). This is denoted the Swain-Schaad relationship. This study examines the breakdown of the Swain Schaad relationship as criterion for tunneling. The semiclassical (no tunneling) limit used hereto (e.g., 3.34, for H/T to D/T kinetic isotope effects), was based on simple theoretical considerations of a diatomic cleavage of a stable covalent bond, for example, a C-H bond. Yet, most experimental evidence for a tunneling contribution has come from breakdown of those relationship for a secondary hydrogen, that is, not the hydrogen whose bond is being cleaved but its geminal neighbor. Furthermore, many of the reported experiments have been mixed-labeling experiments, in which a secondary H/T kinetic isotope effect was measured for C-H cleavage, while the D/T secondary effect accompanied C-D cleavage. In experiments of this type, the breakdown of the Swain-Schaad relationship indicates both tunneling and the degree of coupled motion between the primary and secondary hydrogens. We found a new semiclassical limit (e.g., 4.8 for H/T to D/T kinetic isotope effects), whose breakdown can serve as a more reliable experimental evidence for tunneling in this common mixed-labeling experiment. We study the tunneling contribution to C-H bond activation, for which many relevant experimental and theoretical data are available. However, these studies can be applied to any hydrogen-transfer reaction. First, an extension of the original approach was applied, and then vibrational analysis studies were carried out for a model system (the enzyme alcohol dehydrogenase). Finally, the effect of complex kinetics on the observed Swain-Schaad relationship was examined. All three methods yield a new semiclassical limit (4.8), above which tunneling must be considered. Yet, it was found that for many cases the original, localized limit (3.34), holds fairly well. For experimental results that are between the original and new limits (within statistical errors), several methods are suggested that can support or exclude tunneling. These new and clearer criteria provide a basis for future applications of the Swain-Schaad relationship to demonstrate tunneling in complex systems. PMID- 11942823 TI - Temperature-dependent isotope effects in soybean lipoxygenase-1: correlating hydrogen tunneling with protein dynamics. AB - The hydrogen-atom transfer in soybean lipoxygenase-1 (SLO) exhibits a large kinetic isotope effect on k(cat) (KIE = 81) near room temperature and a very weak temperature dependence (E(act) = 2.1 kcal/mol). These properties are consistent with H small middle dot transfer that occurs entirely by a tunneling event. Mutants of SLO were prepared, and the temperature dependence of the KIE was measured, to test for alterations in the tunneling behavior. All mutants studied exhibit KIEs of similar, large magnitude at 30 degrees C, despite an up to 3 orders of magnitude change in k(cat). E(act) for two of the mutants (Leu(754) --> Ala, Leu(546) --> Ala) is larger than for wild-type (WT), and the KIE becomes slightly more temperature dependent. In contrast, Ile(553) --> Ala exhibits k(cat) and E(act) parameters similar to wild-type soybean lipoxygenase-1 (WT-SLO) for protiated substrate; however, the KIE is markedly temperature dependent. The behavior of the former two mutants could reflect increased reorganization energies (lambda), but the behavior of the latter mutant is inconsistent with this description. We have invoked a full H* tunneling model (Kuznetsov, A. M.; Ulstrup, J. Can. J. Chem. 1999, 77, 1085-1096) to explain the temperature dependence of the KIE, which is indicative of the extent to which distance sampling (gating) modulates hydrogen transfer. WT-SLO exhibits a very small E(act) and a nearly temperature-independent KIE, which was modeled as arising from a compressed hydrogen transfer distance with little modulation of the hydrogen transfer distance. The observations on the Leu(754) --> Ala and Leu(546) --> Ala mutants were modeled as arising from a slightly less compressed active site with greater modulation of the hydrogen transfer distance by environmental dynamics. Finally, the observed behavior of the Ile(553) --> Ala mutant indicates a relaxed active site with extensive involvement of gating to facilitate hydrogen transfer. We conclude that WT-SLO has an active site structure that is well organized to support hydrogen tunneling and that mutations perturb structural elements that support hydrogen tunneling. Modest alterations in active site residues increase lambda and/or increase the hydrogen transfer distance, thereby affecting the probability that tunneling can occur. These studies allow the detection and characterization of a protein-gating mode in catalysis. PMID- 11942824 TI - Alteration of the selectivity of DNA cleavage by a deglycobleomycin analogue containing a trithiazole moiety. AB - The bleomycin (BLM) group of antitumor antibiotics effects DNA cleavage in a sequence-selective manner. Previous studies have indicated that the metal-binding and bithiazole moieties of BLM are both involved in the binding of BLM to DNA. The metal-binding domain is normally the predominant structural element in determining the sequence selectivity of DNA binding, but it has been shown that replacement of the bithiazole moiety with a strong DNA binder can alter the sequence selectivity of DNA binding and cleavage. To further explore the mechanism by which BLM and DNA interact, a trithiazole-containing deglycoBLM analogue was synthesized and tested for its ability to relax supercoiled DNA and cleave linear duplex DNA in a sequence-selective fashion. Also studied was cleavage of a novel RNA substrate. Solid-phase synthesis of the trithiazole deglycoBLM A(5) analogue was achieved using a TentaGel resin containing a Dde linker and elaborated from five key intermediates. The ability of the resulting BLM analogue to relax supercoiled DNA was largely unaffected by introduction of the additional thiazole moiety. Remarkably, while no new sites of DNA cleavage were observed for this analogue, there was a strong preference for cleavage at two 5'-GT-3' sites when a 5'-(32)P end-labeled DNA duplex was used as a substrate. The alteration of sequence selectivity of cleavage was accompanied by some decrease in the potency of DNA cleavage, albeit without a dramatic diminution. In common with BLM, the trithiazole analogue of deglycoBLM A(5) effected both hydrolytic cleavage of RNA in the absence of added metal ion and oxidative cleavage in the presence of Fe(2+) and O(2). In comparison with BLM A(5), the relative efficiencies of hydrolytic cleavage at individual sites were altered. PMID- 11942825 TI - Folding disulfide-containing proteins faster with an aromatic thiol. AB - The traditional method for in vitro folding of disulfide-containing proteins is slow and involves a redox buffer of glutathione and glutathione disulfide. To increase the folding rate and to gain insight into the folding process, we replaced glutathione, an aliphatic thiol, with a commercially available aromatic thiol, 4-mercaptobenzeneacetate (1). Aromatic thiol 1 was selected due to its enhanced nucleophilicity and its enhanced leaving-group ability relative to glutathione at pH 7.7. To demonstrate the advantages of 1, the folding of reduced and scrambled RNase A at pH 7.0 and 7.7 in the presence of 1 and glutathione was investigated. For each set of folding conditions, the optimum concentration of each thiol was initially determined and then the folding rates in the presence of each thiol were measured concurrently. In all cases examined, the folding rate enhancement with the aromatic thiol was 5- 6-fold. Furthermore, under similar conditions folding rates were almost identical with either reduced or scrambled RNase A. In addition the 5-6-fold folding rate enhancement varied only slightly with pH, 7.0 vs 7.7. PMID- 11942826 TI - A comparative photomechanistic study (spin trapping, EPR spectroscopy, transient kinetics, photoproducts) of nucleoside oxidation (dG and 8-oxodG) by triplet excited acetophenones and by the radicals generated from alpha-oxy-substituted derivatives through Norrish-type I cleavage. AB - The photooxidation of 2'-deoxyguanosine (dG) and its derivative 8-oxo-7,8-dihydro 2'-deoxyguanosine (8-oxodG) by a series of acetophenones (AP-X) and benzophenone (BP) has been studied. The favorable absorption characteristics of the benzoyl chromophore enables time-resolved spectroscopy of the triplet ketones to assess their quenching kinetics by dG and 8-oxodG. Whereas the photolysis of acetophenone (AP), 2-acetoxyacetophenone (AP-OAc), and benzophenone (BP) does not produce radicals (group A ketones), the oxymethyl-substituted derivatives 2 hydroxyacetophenone (AP-OH) and 2-tert-butoxyacetophenone (AP-O(t)Bu) lead to carbon-centered radicals by alpha cleavage (group B ketones). For the latter ketones, this was confirmed by EPR studies with the spin trap 5,5 dimethylpyrroline N-oxide (DMPO) and by their triplet lifetimes that were shorter than those for the unsubstituted acetophenone. Both groups of ketones photooxidize dG and 8-oxodG; the oxidation products are spiroiminodihydantoin and guanidine-releasing products (GRP) in the case of dG and AP-OH also 8-oxodG. In the presence of O(2), the photooxidation by the group A ketones is efficient at high dG or 8-oxodG concentrations, whereas the group B ketones photooxidize dG and 8-oxodG also at low substrate concentrations. These results imply that peroxyl radicals are responsible for the photooxidation by the group B ketones, which are formed by alpha cleavage of the triplet ketone and subsequent O(2) trapping of the carbon-centered radicals. At higher dG concentrations, direct electron transfer from dG to the triplet ketone, as observed for the group A ketones, competes with the radical activity. PMID- 11942827 TI - Low-temperature photosensitized oxidation of a guanosine derivative and formation of an imidazole ring-opened product. AB - An organic-soluble guanosine derivative, 2',3',5'-O-(tert butyldimethylsilyl)guanosine (1), was prepared and its photosensitized oxidation was carried out in several solvents at various temperatures. Singlet oxygen is the reactive oxidizing agent responsible for this reaction. Neither an endoperoxide nor a dioxetane intermediate was detected by low-temperature NMR even at -78 degrees C. A product (A) with an oxidized imidazole ring was the only major product detected at room temperature; this compound could be isolated by low-temperature column chromatography and was characterized by (1)H and (13)C and mass spectroscopy. CO(2) was the other major product. A small amount of the corresponding 8-oxo-7,8-dihydroguanosine derivative B was detected during the initial stage of the photooxidation and was shown to be intermediate in the formation of two products of extensive degradation, C and D. Reaction of 1 with the singlet oxygen analogues 4-methyl-1,2,4-triazoline-3,5-dione (MTAD) and 4 phenyl-1,2,4-triazoline-3,5-dione (PTAD) gave products consistent with a proposed mechanism involving the rearrangement of an initially formed endoperoxide to give A and B from reaction of 1 with singlet oxygen. PMID- 11942828 TI - Cooperative binding interactions of glycopeptide antibiotics. AB - Glycopeptide antibiotics of the vancomycin group bind to bacterial cell wall analogue precursors, and typically also form dimers. We have studied the interplay between these two sets of noncovalent bonds formed at separate interfaces. Indole-2-carboxylic acid (L) forms a set of hydrogen bonds to the glycopeptide antibiotic chloroeremomycin (CE) that are analogous to those formed by N-Ac-D-Ala. The ligand/CE dimer interactions (in L/CE/CE/L) are shown to occur with positive cooperativity and structural tightening at the dimer interface. From theoretical considerations and from other data, it is inferred, but not proven, that in the exercise of positive cooperativity, the interface that will be tightened to the greatest degree is the one that lies in the shallowest free energy well. PMID- 11942829 TI - A fluorescent anion sensor that works in neutral aqueous solution for bioanalytical application. AB - Anion recognition and anion sensing are of interest because anions play many important roles in living organisms. Most currently known anion sensors work only in organic solution, but sensors for biological applications are required to function in neutral aqueous solution. We have designed and synthesized a novel fluorescent sensor for anions. The sensor molecule 1-Cd(II) contains 7-amino-4 trifluoromethylcoumarin as a fluorescent reporter and Cd(II)-cyclen (1,4,7,10 tetraazacyclododecane) as an anion host. In neutral aqueous solution, Cd(II) of 1 Cd(II) is coordinated by the four nitrogen atoms of cyclen and the aromatic amino group of coumarin. When various anions are added to 100 mM HEPES buffer solution (pH 7.4) containing 1-Cd(II), the aromatic amino group of coumarin is displaced from Cd(II), causing a change of the excitation spectrum. While pyrophosphate and citrate were detected with high sensitivity, fluoride and perchlorate produced no response. Among organic anions, ATP and ADP gave strong signals, while cAMP showed little signal. By utilizing the different affinities of the sensor for AMP and cAMP, the activity of phosphodiesterase, which cleaves cyclic nucleotide, was monitored in real-time. The sensor should have many biochemical and analytical applications and the sensing principle should be widely applicable to the sensing of other molecules. PMID- 11942830 TI - The effect of macromolecular architecture in nanomaterials: a comparison of site isolation in porphyrin core dendrimers and their isomeric linear analogues. AB - The influence of macromolecular architecture on the physical properties of polymeric materials has been studied by comparing poly(benzyl ether) dendrons with their exact linear analogues. The results clearly confirm the anticipation that dendrimers are unique when compared to other architectures. Physical properties, from hydrodynamic volume to crystallinity, were shown to be different, and in a comparative study of core encapsulation in macromolecules of different architecture, energy transduction from the polymer backbone to a porphyrin core was shown to be different for dendrimers as compared to that of isomeric four- or eight-arm star polymers. Fluorescence excitation revealed strong, morphology dependent intramolecular energy transfer in the three macromolecular isomers investigated. Even at high generations, the dendrimers exhibited the most efficient energy transfer, thereby indicating that the dendritic architecture affords superior site isolation to the central porphyrin it surrounds. PMID- 11942831 TI - Stereoselective total syntheses and reassignment of stereochemistry of the freshwater cyanobacterial hepatotoxins cylindrospermopsin and 7 epicylindrospermopsin. AB - A stereoselective total synthesis of the structure 1 proposed for the freshwater cyanobacterial heptatotoxin cylindrospermopsin has been accomplished in approximately 30 operations starting from commercially available 4 methoxypyridine. Utilizing methodology developed by Comins, the tetrasubstituted piperidine A-ring unit of the hepatotoxin was efficiently constructed. The two remaining stereocenters in the natural product were then set by a stereospecific intramolecular N-sulfinylurea Diels-Alder cyclization/Grignard ring opening/allylic sulfoxide [2,3]-sigmatropic rearrangement sequence previously developed in these laboratories, leading to key intermediate 29. The stereochemical assignment of alcohol 29, which contains all six of the stereogenic centers of the natural product, was confirmed by an X-ray crystal structure determination of a derivative. Installation of the D-ring uracil moiety was effected by using our new methodology developed for this purpose, and construction of the C-ring guanidine completed the total synthesis of racemic structure 1. However, the (1)H NMR data for this compound do not match that of cylindrospermopsin, but instead agree with the data reported for 7 epicylindrospermopsin, a minor toxic metabolite that co-occurs with cylindrospermopsin. Therefore, we propose a revision of the stereochemical assignments of these natural products such that cylindrospermopsin is now represented as structure 2 and 7-epicylindrospermopsin is 1. This reassignment was further confirmed by Mitsunobu inversion of the C-7 alcohol 51 to epimer 52, and conversion of this compound to tetracyclic diol 57, which has previously been transformed to cylindrospermopsin (2). PMID- 11942832 TI - Polymeric complexes of silver(I) with diphosphine ligands: self-assembly of a puckered sheet network structure. AB - The syntheses and structures of polymeric silver(I)-diphosphine complexes are reported, in which the silver(I) center is surrounded by 1, 2, or 3 phosphorus atoms. When rigid diphosphine ligands are used in combination with weakly coordinating anions, linear polymers are obtained that contain both diphosphine and anion bridges. However, with excess of a diphosphine with a long, flexible, spacer group, a remarkable puckered sheet structure, comprised of fused giant 54 membered rings, is obtained that is a coordination polymer analogue of laminated materials such as micas and clays. The polymeric chain and sheet structures may be considered to be formed by ring-opening polymerization of cyclic precursors. PMID- 11942833 TI - Variable-temperature nuclear magnetic resonance spectroscopy allows direct observation of carboxylate shift in zinc carboxylate complexes. AB - Tetranuclear complexes [Zn(4)(bdmap)(2)(OOCR)(6)] 1 (R = Me) and 2 (R = Et), where Hbdmap = 1,3-bis(dimethylamino)-2-propanol, were prepared from zinc carboxylates and Hbdmap in tetrahydrofuran (THF). The solid-state structures of isomers 1a and 2a consist of two pairs of zinc atoms, each bridged by two mu-1,2 and one mu-1,1 carboxylate ligands. Two pairs are connected by two tridentate bdmap ligands with oxygen acting as a bridging donating atom. The complexes retain the tetranuclear structure in solution and two dynamic processes are observed from variable-temperature (1)H and (13)C NMR spectra. A low-temperature process (LT dynamics) observed already below 200 K is a coalescence of the mu-1,2 and the mu-1,1 resonances to a single resonance. An additional dynamic process (HT dynamics) is observed above 247 K (1) and 263 K (2), leading to a coalescence of two dimethylamino resonances. Both dynamic processes are rationalized by a mechanism involving changes in the carboxylate coordination mode termed as carboxylate shift. The LT dynamics is ascribed to interconversions of a single mu 1,2 and a single mu-1,1 carboxylate ligation by rotations of 60 degrees. The interconversions involve all carboxylate ligands in 1 and 2. The HT dynamics is ascribed to the exchange of the coordinating geometries of two carboxylate bridged zinc atoms. We propose a mechanism that starts with a cleavage of the Zn N coordination bond. The resulting coordinatively unsaturated zinc atom acquires an additional oxygen donor atom by carboxylate shift of mu-1,2 carboxylate to mu 1,1 mode. The activation parameters (DeltaH values in kilocalories per mole, DeltaS values in calories per mole per kelvin) were determined by line-shape analysis of VT NMR spectra: for 1 in THF-d(8), DeltaH(LT) = 8.1(3), DeltaS(LT) = 12(2), DeltaH(HT) = 17.9(2), DeltaS(HT) = 14(1); for 1 in CDCl(3), DeltaH(HT) = 13.6(5), DeltaS(HT) = 3(3); for 1 in CD(2)Cl(2), DeltaH(HT) = 9.9(3), DeltaS(HT) = -8(2); for 2 in THF-d(8), DeltaH(LT) = 11(1), DeltaS(LT) = -5(3), DeltaH(HT) = 19.6(5), DeltaS(HT) = 18(3). Polymeric [Zn(4)(bdmap)(2)(OOCMe)(6)](n) 1-catena crystallizes from a dichloromethane solution of 1. In 1-catena, the zinc atoms are linked into a chain through mu-1,2 and mu-1,1 acetate alternated by mu-1,2 acetate and bdmap. PMID- 11942834 TI - Gold(I) macrocycles and topologically chiral [2]catenanes. AB - The design and synthesis of a new type of topologically chiral [2]catenane is reported. The compounds are formed easily by self-assembly on reaction of the oligomeric digold(I) diacetylide precursor complex [[4-BrC(6)H(4)CH(4 C(6)H(4)OCH(2)CCAu)(2)](n)] with diphosphine ligands. Reactions with the diphosphines PP = bis(diphenylphosphinophoshino)acetylene, trans-1,2 bis(diphenylphosphino)ethylene, bis(diphenylphosphino)ethane, and 1,1' bis(diphenylphosphino)ferrocene yield simple ring complexes [4-BrC(6)H(4)CH(4 C(6)H(4)OCH(2)CCAu)(2)(mu-PP)] as the only products, since the spacer groups in the diphosphines are not long enough or are too bulky to allow catenane formation. Reaction with PP = bis(diphenylphosphino)propane or bis(diphenylphosphino)butane gave [2]catenane complexes [[4-BrC(6)H(4)CH(4 C(6)H(4)OCH(2)CCAu)(2)(mu-PP)](2)], whose structures are confirmed crystallographically. The macrocyclic ring compounds have C(s) symmetry but, as a result of the presence of the unsymmetrical "hinge group" 4-BrC(6)H(4)CH, the [2]catenanes have C(2) symmetry and so are topologically chiral. In favorable cases, the formation of the [2]catenane can be proved by NMR spectroscopy since catenane formation leads to nonequivalence of most ring atoms. The formation of the [2]catenanes was successfully predicted based on the conformation of the precursor bis(phenol), and it is argued that the methods used should be more generally applicable to the synthesis of functionally substituted supermolecules of interest for application in molecular devices. PMID- 11942835 TI - Pb(3)F(5)NO(3), a cationic layered material for anion-exchange. AB - Our research involves the development of new cationic materials for anion-based applications. We report the solvothermal synthesis and characterization of Pb(3)F(5)NO(3), a new layered lead fluoride material that, unlike the majority of layered and open-framework materials, is cationic in charge. The structure consists of polyhedral lead centers connected by doubly and triply bridging fluoride groups. We quantitatively exchanged the interlamellar nitrate groups of Pb(3)F(5)NO(3) for dichromate, under ambient aqueous conditions. Nuclear magnetic resonance and UV-vis spectroscopy show the reaction proceeds to 61.0% completion in several days. The material is also stable to 450 degrees C, which is vastly superior to organic resins that are still the standard for anion-exchange. The presence of extraframework anions also opens up other potentially unique anion based properties, such as new catalytic reactions, anion intercalation, or growth of anionic clusters within the void spaces of the cationic material. PMID- 11942836 TI - Kinetic isotope effects in cycloreversion of rhenium (V) diolates. AB - Cycloreversion of 4-methoxystyrene from the corresponding Tp'Re(O)(diolato) complex (Tp' = hydrido-tris-(3,5-dimethylpyrazolyl)borate) was measured competitively for various isotopomers at 103 degrees C. Primary ((12)C/(13)C) and secondary ((1)H/(2)H) kinetic isotope effects were determined. The primary KIEs were k(12C)/k(13C) = 1.041 +/- 0.005 at the alpha position and 1.013 +/- 0.006 at the beta position. Secondary KIEs were k(H)/k(D) = 1.076 +/- 0.005 at the alpha position and 1.017 +/- 0.005 at the beta position. Computational modeling (B3LYP/LACVP+) located a transition state for concerted cycloreversion of styrene from TpRe(O)(OCH(2)CHPh) exhibiting dramatically different C-O bond lengths. A Hammett study on cycloreversions of substituted styrenes from a series of Tp'Re(O)(diolato) showed dichotomous behavior for electron donors and electron withdrawing groups as substituents: rho = -0.65 for electron donors, but rho = +1.13 for electron-withdrawing groups. The data are considered in light of various mechanistic proposals. While the extrusion of 4-methoxystyrene is concluded to be a highly asynchronous concerted reaction, the Hammett study reflects a likelihood that multiple reaction mechanisms are involved. PMID- 11942837 TI - CdS nanoparticles modified to chalcogen sites: new supramolecular complexes, butterfly bridging, and related optical effects. AB - All present approaches to surface modification of nanoparticles (NPs) with organic ligands exploit metal (cadmium) sites as anchor points. To obtain efficient interaction of NP surface with p-orbitals of organic chromophores, we utilize the chalcogen (sulfur) sites on the NP surface. These sites present several advantages stemming from a stronger interaction of their atomic orbitals with both modifier and NP core. The chalcogen modification of CdS was achieved by using a mixed ligand (2,2'-bipyridyl-N,N')(malonato-O,O')-copper(II) monohydrate complex. The weak monodentate ligands (water) are replaced by a copper-sulfur bond during the modification reaction. The structure of the product was investigated by optical spectroscopy, electron spin resonance, and nuclear magnetic resonance. The modified NP can be described as a few tens (<40) of (2,2' bipyridyl-N,N')(malonato-O,O')-copper units attached to the CdS core. Steady state and time-resolved luminescence measurements, molecular orbital calculations, and UPS data indicate that delocalized surface states enveloping the surface chalcogen atoms of NP, transition metal, and p-orbitals of the bipyridine ligand are present in the synthesized species. The delocalized states are made possible due to the bridging of p-levels of sulfur and pi-orbitals of bipyridine by butterfly d-orbitals of the transition metal atom placed between them. Chalcogen-modified NP can be considered as a new member of the family of supramolecular compounds based on transition metal complexes. Both NP and metal complex parts of the prepared supramolecules are very versatile structural units, and new molecular constructs of similar design, in which quantum effects of NPs are combined with optical properties of transition metal complexes, can be obtained with different NPs and metal complexes. PMID- 11942838 TI - Functional mimic of dioxygen-activating centers in non-heme diiron enzymes: mechanistic implications of paramagnetic intermediates in the reactions between diiron(II) complexes and dioxygen. AB - Two tetracarboxylate diiron(II) complexes, [Fe(2)(mu O(2)CAr(Tol))(2)(O(2)CAr(Tol))(2)(C(5)H(5)N)(2)] (1a) and [Fe(2)(mu O(2)CAr(Tol))(4)(4-(t)BuC(5)H(4)N)(2)] (2a), where Ar(Tol)CO(2)(-) = 2,6-di(p tolyl)benzoate, react with O(2) in CH(2)Cl(2) at -78 degrees C to afford dark green intermediates 1b (lambda(max) congruent with 660 nm; epsilon = 1600 M(-1) cm(-1)) and 2b (lambda(max) congruent with 670 nm; epsilon = 1700 M(-1) cm(-1)), respectively. Upon warming to room temperature, the solutions turn yellow, ultimately converting to isolable diiron(III) compounds [Fe(2)(mu-OH)(2)(mu O(2)CAr(Tol))(2)(O(2)CAr(Tol))(2)L(2)] (L = C(5)H(5)N (1c), 4-(t)BuC(5)H(4)N (2c)). EPR and Mossbauer spectroscopic studies revealed the presence of equimolar amounts of valence-delocalized Fe(II)Fe(III) and valence-trapped Fe(III)Fe(IV) species as major components of solution 2b. The spectroscopic and reactivity properties of the Fe(III)Fe(IV) species are similar to those of the intermediate X in the RNR-R2 catalytic cycle. EPR kinetic studies revealed that the processes leading to the formation of these two distinctive paramagnetic components are coupled to one another. A mechanism for this reaction is proposed and compared with those of other synthetic and biological systems, in which electron transfer occurs from a low-valent starting material to putative high-valent dioxygen adduct(s). PMID- 11942840 TI - Template synthesis of metal nanowires containing monolayer molecular junctions. AB - Metal nanowires containing in-wire monolayer junctions of 16-mercaptohexanoic acid were made by replication of the pores of 70 nm diameter polycarbonate track etch membranes. Au was electrochemically deposited halfway through the 6 microm long pores and a self-assembled monolayer (SAM) of 16-mercaptohexadecanoic acid was adsorbed on top. A thin layer of Au was then electrolessly grown to form a metal cap separated from the bottom part of the wire by the SAM. Electron micrographs showed that the bottom and top metal segments were separated by an approximately 2 nm thick organic monolayer. Current-voltage measurements of individual nanowires confirmed that the organic monolayer could be contacted electrically on the top and bottom by the metal nanowire segments without introducing electrical short circuits that penetrate the monolayer. The values of the electrical properties for zero-bias resistance, current density, and breakdown field strength were within the ranges expected for a well-ordered alkanethiol SAM of this thickness. PMID- 11942839 TI - Investigation of the electron-transfer mechanism by cross-linking between Zn substituted myoglobin and cytochrome b(5). AB - We have investigated the photoinduced electron transfer (ET) in the 1:1 cross linked complex (CL-ZnMb/b(5)) formed by a cross-linking reagent, EDC, between Zn substituted myoglobin (ZnMb) and cytochrome b(5) (Cytb(5)) to reveal the mechanism of the inter-protein ET reactions under the condition of multiple encounter complexes. A variety of the ZnMb-Cytb(5) orientations was suggested because of failure to identify the single and specific cross-linking site on Cytb(5) by the peptide-mapping analysis using mass spectrometry. In CL-ZnMb/b(5), a laser pulse generates the triplet excited state of the ZnMb domain ((3)ZnMb()), which can transfer one electron to the Cytb(5) domain. The decay kinetics of (3)ZnMb() in CL-ZnMb/b(5) consists of a facile power-law ET phase to Cytb(5) domain ( approximately 30%) and a slower single-exponential phase ( approximately 70%). The application of the Marcus equation to this power-law phase indicates that CL-ZnMb/b(5) has a variety of ZnMb-Cytb(5) orientations for the facile ET in which the distance between the redox centers (D-A distance) is distributed over 13-20 A. The single-exponential phase in the (3)ZnMb() decay kinetics of CL ZnMb/b(5) is similar to the intrinsic decay of (3)ZnMb() in its rate constant, 65 s(-)(1). This implies that the ET is impeded in about 70% of the total ZnMb Cytb(5) orientations due to the D-A distance larger than 20 A. Combined with the results of the Brownian dynamics simulations for the encounter complexes, the overall bimolecular ET rate, k(app), can be reproduced by the sum of the ET rates for the minor encounter complexes of which D-A distance is less than 20 A. On the other hand, the encounter complexes with longer D-A distance, which are the majority of the encounter complexes between ZnMb and Cytb(5), have little contribution to the overall bimolecular ET rate. These observations experimentally demonstrate that ZnMb forms a variety of encounter complexes with Cytb(5), among which a minor set of the complexes with the shorter D-A distance (< approximately 20 A) regulates the overall bimolecular ET between the proteins. PMID- 11942841 TI - Proton transfer reactions on semiconductor surfaces. AB - The concept of proton affinity on semiconductor surfaces has been explored through an investigation of the chemistry of amines on the Ge(100)-2 x 1, Si(100) 2 x 1, and C(100)-2 x 1 surfaces. Multiple internal reflection Fourier transform infrared (MIR-FTIR) spectroscopy, temperature programmed desorption (TPD), and density functional theory (DFT) calculations were used in the studies. We find that methylamine, dimethylamine, and trimethylamine undergo molecular chemisorption on the Ge(100)-2 x 1 surface through the formation of Ge-N dative bonds. In contrast, primary and secondary amines react on the Si(100)-2 x 1 surface via N-H dissociation. Since N-H dissociation of amines at semiconductor surfaces mimics a proton-transfer reaction, the difference in chemical reactivities of the Ge(100)-2 x 1 and Si(100)-2 x 1 surfaces toward N-H dissociation can be interpreted as a decrease of proton affinity down a group in the periodic table. The trend in proton affinities of the two surfaces is explained in terms of thermodynamics and kinetics. Solid-state effects on the C(100)-2 x 1 surface and the surface proton affinity concept are discussed based on our theoretical predictions. PMID- 11942842 TI - A mechanism from quantum chemical studies for methane formation in methanogenesis. AB - The mechanism for methane formation in methyl-coenzyme M reductase (MCR) has been investigated using the B3LYP hybrid density functional method and chemical models consisting of 107 atoms. The experimental X-ray crystal structure of the enzyme in the inactive MCR(ox1)(-)(silent) state was used to set up the initial model structure. The calculations suggest a mechanism not previously proposed, in which the most remarkable feature is the formation of an essentially free methyl radical at the transition state. The reaction cycle suggested starts from a Michaelis complex with CoB and methyl-CoM coenzymes bound and with a squareplanar coordination of the Ni(I) center in the tetrapyrrole F(430) prosthetic group. In the rate-limiting step the methyl radical is released from methyl-CoM, induced by the attack of Ni(I) on the methyl-CoM thioether sulfur. In this step, the metal center is oxidized from Ni(I) to Ni(II). The resulting methyl radical is rapidly quenched by hydrogen-atom transfer from the CoB thiol group, yielding the methane molecule and the CoB radical. The estimated activation energy is around 20 kcal/mol, which includes a significant contribution from entropy due to the formation of the free methyl. The mechanism implies an inversion of configuration at the reactive carbon. The size of the inversion barrier is used to explain the fact that CF(3)-S-CoM is an inactive substrate. Heterodisulfide CoB-S-S-CoM formation is proposed in the final step in which nickel is reduced back to Ni(I). The suggested mechanism agrees well with experimental observations. PMID- 11942843 TI - Measurement of long-range cross-correlation rates using a combination of single- and multiple-quantum NMR spectroscopy in one experiment. AB - A method is described to determine long-range cross-correlations between the modulations of an anisotropic chemical shift (e.g., of a C' carbonyl carbon in a protein) and the fluctuations of a weak long-range dipolar interaction (e.g., in cross-correlation between the same C' carbonyl and the H(N) proton of the neighboring amide group). Such long-range correlations are difficult to measure because the corresponding long-range scalar couplings are so small that Redfield's secular approximation is often violated. The method, which combines features of single- and double-quantum NMR spectroscopy, allows one to cancel the effects of dominant short-range dipolar interactions (e.g., between the CSA of the amide nitrogen N and the dipolar coupling to its attached proton H(N)) and is designed so that the secular approximation is rescued even if the scalar coupling between the long-range dipolar coupling partners is very small. The cross correlation rates thus determined in ubiquitin cover a wide range because of local motions and variations of the CSA tensors. PMID- 11942844 TI - Oxygen-transfer reactions between 3d transition metals and N(2)O and NO(2). AB - Density functional calculations have been performed to describe reactions of ground-state 3d transition metal atoms (Sc-Ni) with N(2)O and NO(2) molecules. From the analysis of the calculated reaction surfaces, a general reaction mechanism evolved. The reactions are initiated by electron transfer from metal to the oxidant molecule, which weakens the N-O bond and facilitates an O(-)((2)P) abstraction. 4s-3d hybridization taking place in the metal electronic structure plays an essential role in the net 4s(beta) electron transfer from the metal atom to the nitrogen-oxide molecule. These key steps contribute to connect the reactant and product channels on a single potential energy surface. The calculations revealed that reaction with NO(2) yields stable oxo-nitrosyl insertion products, and their equilibrium structural properties can be understood by inspecting the 4pi* metal-oxide orbital occupancies. Correlation is obtained between the metal 3d ionization energies and the reaction rates as well as activation energies. This correlation provides additional support for the reaction mechanism called electron-transfer-assisted oxygen abstraction. This novel mechanism exhibits the basic features of the simple electron transfer and direct abstraction kinetic models and sheds new light on the so-called resonance interaction model as well. PMID- 11942845 TI - Photoisomerization and photodissociation of toluene in molecular beam. AB - The photodissociation of isotope-labeled toluene C(6)H(5)CD(3) and C(6)H(5)(13)CH(3) molecules at 6.4 eV under collision-free conditions was studied in separate experiments by multimass ion imaging techniques. In addition to the major dissociation channels, C(6)H(5)CD(3) --> C(6)H(5)CD(2) + D and C(6)H(5)CD(3) --> C(6)H(5) + CD(3), the respective photofragments CD(2)H, CDH(2), and CH(3) and their heavy fragment partners C(6)H(4)D, C(6)H(3)D(2), and C(6)H(2)D(3) were observed from C(6)H(5)CD(3) dissociation. Photofragments (13)CH(3) and CH(3), and their heavy fragment partners C(6)H(5) and (13)CC(5)H(5), were also observed from C(6)H(5)(13)CH(3) dissociation. Our results show that 25% of the excited toluene isomerizes to a seven-membered ring (cycloheptatriene) and then rearomatizes prior to dissociation. The isomerization pathway competes with direct C-C bond and C-H bond dissociation. The significance of this isomerization is that the carbon atoms and hydrogen atoms belonging to the alkyl group are involved in an exchange with those atoms in the aromatic ring during isomerization. The dissociation rate of toluene at 193 nm is measured to be (1.17 +/- 0.1) x 10(6) s(-)(1). PMID- 11942846 TI - Reactions of carbocations with unsaturated hydrocarbons: electrophilic alkylation or hydride abstraction? AB - Benzhydryl cations were used as reference electrophiles to determine the hydride donor reactivities of unsaturated hydrocarbons. The kinetics of the reactions were followed by UV-vis spectroscopy and conductivity measurements, and it was found that the second-order rate constants for the hydride transfer processes were almost independent of the solvents or counterions employed. The rate constants correlate linearly with the previously published empirical electrophilicity parameters E of the benzhydrylium ions. Therefore, the linear free energy relationship log k(20 degrees C) = s(E + N) could be employed to characterize the hydride reactivities of the hydrocarbons by the nucleophilicity parameters N and s. The similarity of the slopes s for hydride donors and pi nucleophiles allows a direct comparison of the reactivities of these different functional groups based on their nucleophilicity parameters N. Since nucleophilicity parameters of -5 < N < 0 have been found for a large variety of allylic and bisallylic hydride donors, a rule of thumb is derived that hydride transfer processes may compete with carbon-carbon bond-forming reactions when carbocations are combined with olefins of pi-nucleophilicity N < 0. PMID- 11942847 TI - Rate-equilibrium relationships in hydride transfer reactions: the role of intrinsic barriers. AB - A literature survey on the kinetics of hydride abstractions from CH-groups by carbocations reveals a general phenomenon: Variation of the hydride acceptor affects the rates of hydride transfer to a considerably greater extent than an equal change of the thermodynamic driving force caused by variation of the hydride donor. The origin of this relationship was investigated by quantum chemical calculations on various levels of ab initio and DFT theory for the transfer of an allylic hydrogen from 1-mono- and 1,1-disubstituted propenes (XYC=CH-CH(3)) to the 3-position of 1-mono- and 1,1-disubstituted allyl cations (XYC=CH-CH(2)(+)). The discussion is based on the results of the MP2/6 31+G(d,p)//RHF/6-31+G(d,p) calculations. Electron-releasing substituents X and Y in the hydride donors increase the exothermicity of the reaction, while electron releasing substituents in the hydride acceptors decrease exothermicity. In line with Hammond's postulate, increasing exothermicity shifts the transition states on the reaction coordinate toward reactants, as revealed by the geometry parameters and the charge distribution in the activated complexes. Independent of the location of the transition state on the reaction coordinate, a value of 0.72 is found for Hammond-Leffler's alpha = deltaDeltaG/deltaDelta(r)G degrees when the hydride acceptor is varied, while alpha = 0.28 when the hydride donor is varied. The value of alpha thus cannot be related with the position of the transition state. Investigation of the degenerate reactions XYC=CH-CH(3) + XYC=CH CH(2)(+) indicates that the migrating hydrogen carries a partial positive charge in the transition state and that the intrinsic barriers increase with increasing electron-releasing abilities of X and Y. Substituent variation in the donor thus influences reaction enthalpy and intrinsic barriers in the opposite sense, while substituent variation in the acceptor affects both terms in the same sense, in accord with the experimental findings. Marcus theory is employed to treat these effects quantitatively. PMID- 11942848 TI - A general method for determining the electron self-exchange rates of blue copper proteins by longitudinal NMR relaxation. AB - A general NMR method is presented that allows a precise determination of the second-order rate constant, k(ese), for the electron self-exchange in blue copper proteins, from the longitudinal relaxation rates of the nuclei in the protein. The method relies on the use of partly oxidized (paramagnetic) samples of the protein. In contrast to previous NMR approaches for the determination of electron self-exchange rates, the applicability of the method extends beyond the slow exchange limit, k(ese)c << R(ip), i = 1, 2, where c is the protein concentration, and R(ip) is the paramagnetic relaxation enhancement of the observed nuclei. PMID- 11942849 TI - How much do enzymes really gain by restraining their reacting fragments? AB - The steric effect, exerted by enzymes on their reacting substrates, has been considered as a major factor in enzyme catalysis. In particular, it has been proposed that enzymes catalyze their reactions by pushing their reacting fragments to a catalytic configuration which is sometimes called near attack configuration (NAC). This work uses computer simulation approaches to determine the relative importance of the steric contribution to enzyme catalysis. The steric proposal is expressed in terms of well defined thermodynamic cycles that compare the reaction in the enzyme to the corresponding reaction in water. The S(N)2 reaction of haloalkane dehalogenase from Xanthobacter autotrophicus GJ10, which was used in previous studies to support the strain concept is chosen as a test case for this proposal. The empirical valence bond (EVB) method provides the reaction potential surfaces in our studies. The reliability and efficiency of this method make it possible to obtain stable results for the steric free energy. Two independent strategies are used to evaluate the actual magnitude of the steric effect. The first applies restraints on the substrate coordinates in water in a way that mimics the steric effect of the protein active site. These restraints are then released and the free energy associated with the release process provides the desired estimate of the steric effect. The second approach eliminates the electrostatic interactions between the substrate and the surrounding in the enzyme and in water, and compares the corresponding reaction profiles. The difference between the resulting profiles provides a direct estimate of the nonelectrostatic contribution to catalysis and the corresponding steric effect. It is found that the nonelectrostatic contribution is about -0.7 kcal/mol while the full "apparent steric contribution" is about -2.2 kcal/mol. The apparent steric effect includes about -1.5 kcal/mol electrostatic contribution. The total electrostatic contribution is found to account for almost all the observed catalytic effect ( approximately -6.1 kcal/mol of the -6.8 calculated total catalytic effect). Thus, it is concluded that the steric effect is not the major source of the catalytic power of haloalkane dehalogenase. Furthermore, it is found that the largest component of the apparent steric effect is associated with the solvent reorganization energy. This solvent-induced effect is quite different from the traditional picture of balance between the repulsive interaction of the reactive fragments and the steric force of the protein. PMID- 11942850 TI - Hydrogen atom abstraction reactions of charged polyaromatic sigma-radicals related to the active intermediates of the enediyne antitumor drugs. AB - Polar effects are demonstrated to play an important role in controlling the reactivity of polyaromatic sigma-radicals that are structurally related to the active intermediates of the enediyne anticancer type antibiotics. This was accomplished by measuring the rate constants of hydrogen atom abstraction for novel, charged dehydroquinolines, dehydroisoquinolines, dehydrobenzenes, and dehydronaphthalenes in the gas phase by using Fourier-transform ion cyclotron resonance mass spectrometry. The reactivity trends observed for these radicals upon hydrogen atom abstraction from tetrahydrofuran and 2-methyltetrahydrofuran, simple models of deoxyribose, do not reflect differences in reaction exothermicities, radical sizes, exact location of the radical site in the ring system, or heteroatom-radical site distances. However, the reactivity trends match the trend in the calculated electron affinities of the radicals. The radicals' different electrophilicities result in variations in the reaction barrier due to different extents of polarization of the transition state. Generally, the reaction efficiencies are the greatest when the formally charged heteroatom is contained within the same ring system as the radical site. In this case, polar effects have the greatest influence on radical reactivity. Hence, insertion of a basic heteroatom (which gets protonated in biological systems) into specific locations in the polyaromatic ring system of the sigma-biradicals, which ultimately cause cleavage of DNA exposed to the enediyne antitumor drugs, should allow tuning of the reactivity of these radicals. PMID- 11942851 TI - Chemical bonding in group III nitrides. AB - We analyze in this article the evolution of the chemical bonding in group III nitrides (MN, M = Al, Ga, In), from the N-N bond dominated small clusters to the M-N bond dominated crystals, with the aim of explaining how the strong multiple bond of N(2) is destabilized with the increase in coordination. The picture that emerges is that of a partially ionic bond in the solid state, which is also present in all the clusters. The covalent N-N bond, however, shows a gradual decrease of its strength due to the charge transfer from the metal atoms. Overall, Al clusters are more ionic than Ga and In clusters, and thus the N-N bond is weakest in them. The nitrogen atom charge is seen to be proportional to the metal coordination, being thus a bond-related property, and dependent on the M-N distance. This explains the behavior observed in previous investigations, and can be used as a guide in predicting the structures and defects on semiconductor quantum dot or thin film devices of these compounds. PMID- 11942852 TI - Reaction path analysis of the "tunable" photoisomerization selectivity of free and locked retinal chromophores. AB - Multiconfigurational second-order perturbation theory computations and reaction path mapping for the retinal protonated Schiff base models all-trans-nona-2,4,6,8 tetraeniminium and 2-cis-nona-2,4,6,8-tetraeniminium cation demonstrate that, in isolated conditions, retinal chromophores exhibit at least three competing excited-state double bond isomerization paths. These paths are associated with the photoisomerization of the double bonds in positions 9, 11, and 13, respectively, and are controlled by barriers that favor the position 11. The computations provide a basis for the understanding of the observed excited-state lifetime in both naturally occurring and synthetic chromophores in solution and, tentatively, in the protein environment. In particular, we provide a rationalization of the excited-state lifetimes observed for a group of locked retinal chromophores which suggests that photoisomerization in bacteriorhodopsin is the result of simultaneous specific "catalysis" (all-trans --> 13-cis path) accompanied by specific "inhibition" (all-trans --> 11-cis path). The nature of the S(1) --> S(0) decay channel associated with the three paths has also been investigated at the CASSCF level of theory. It is shown that the energy surfaces in the vicinity of the conical intersection for the photoisomerization about the central double bond of retinal (position 11) and the two corresponding lateral double bonds (positions 9 and 13) are structurally different. PMID- 11942853 TI - Theoretical study of the mechanism of alkane hydroxylation and ethylene epoxidation reactions catalyzed by diiron bis-oxo complexes. The effect of substrate molecules. AB - The hybrid density functional method B3LYP was used to study the mechanism of the hydrocarbon (methane, ethane, methyl fluoride, and ethylene) oxidation reaction catalyzed by the complexes cis-(H(2)O)(NH(2))Fe(mu-O)(2)(eta(2) HCOO)(2)Fe(NH(2))(H(2)O), I, and cis-(HCOO)(Imd)Fe(mu-O)(2)(eta(2) HCOO)(2)Fe(Imd)(HCOO) (Imd = Imidazole), I_m, the "small" and "medium" model of compound Q of the methane monooxygenase (MMO). The improvement of the model from "small" to "medium" did not change the qualitative conclusions but significantly changed the calculated energetics. As in the case of methane oxidation reported by the authors previously, the reaction of all the substrates studied here is shown to start by coordination of the substrate molecule to the bridging oxygen atom, O(1) of I, an Fe(IV)-Fe(IV) complex, followed by the H-atom abstraction at the transition state III leading to the bound hydroxy alkyl intermediate IV of Fe(III)-Fe(IV) core. IV undergoes a very exothermic coupling of alkyl and hydroxy groups to give the alcohol complex VI of Fe(III)-Fe(III) core, from which alcohol dissociates. The H(b)-atom abstraction (or C-H bond activation) barrier at transition state III is found to be a few kcal/mol lower for C(2)H(6) and CH(3)F than for CH(4). The calculated trend in the H(b)-abstraction barrier, CH(4) (21.8 kcal/mol) > CH(3)F (18.8 kcal/mol) > or = C(2)H(6) (18.5 kcal/mol), is consistent with the C-H(b) bond strength in these substrates. Thus, the weaker the C-H(b) bond, the lower is the H(b)-abstraction barrier. It was shown that the replacement of a H-atom in a methane molecule with a more electronegative group tends to make the H(b)-abstraction transition state less "reactant-like". In contrast, the replacement of the H-atom in CH(4) with a less electronegative group makes the H(b)-abstraction transition state more "reactant-like". The epoxidation of ethylene by complex I is found to proceed without barrier and is a highly exothermic process. Thus, in the reaction of ethylene with complex I the only product is expected to be ethylene oxide, which is consistent with the experiment. PMID- 11942854 TI - Computational studies of tungsten-catalyzed endo-selective cycloisomerization of 4-pentyn-1-ol. AB - Endo- and exo-cycloisomerizations of 4-pentyn-1-ol have been studied computationally with density functional theory, in conjunction with double-zeta and triple-zeta basis sets, both in the absence and in the presence of tungsten carbonyl catalyst. In the absence of the catalyst, both endo- and exo cycloisomerizations have been calculated to have a very high activation barrier of approximately 50-55 kcal/mol and cannot take place. With tungsten pentacarbonyl catalyst, endo-cycloisomerization becomes a complex multiple-step reaction and proceeds with a rate-determining barrier of 26 kcal/mol at the C(alpha) --> C(beta) hydride migration step to form a vinylidene intermediate. The primary role of the tungsten catalyst is to stabilize the vinylidene intermediate, thus lowering the rate-determining barrier. The second important role of the tungsten catalyst in endo-cycloisomerization is to assist the OH hydride migration to C(alpha) by making it a multistep process with small activation barriers. The exo-cycloisomerization with the catalyst still has a high rate-determining barrier of 47 kcal/mol. These findings clearly explain the experimentally observed endo-selectivity in the cycloisomerization of 4-pentyn-1 ol derivatives and support the experimentally proposed mechanism. PMID- 11942855 TI - Structural dependencies of interresidue scalar coupling (h3)J(NC') and donor (1)H chemical shifts in the hydrogen bonding regions of proteins. AB - A study is presented of the structural dependencies for scalar J-coupling and the amide donor (1)H chemical shifts in the hydrogen bonding regions of proteins. An analysis of the interactions between the donor hydrogen and acceptor oxygen orbitals in an N-H...O=C moiety suggests that there are three major structural factors for (15)N-(13)C coupling across hydrogen bonds: (1) the H...O' internuclear separation r(HO)('), (2) the H...O'=C' angle theta(2), and (3) indirect contributions involving the oxygen loan pair electrons should lead to a dependence on the H...O'=C'-N' dihedral angle rho. Density functional theory (DFT) and finite perturbation theory (FPT) were used to obtain the Fermi contact (FC) contributions to interresidue coupling in formamide dimers with systematic variation of these structural parameters. The computed (h3)J(NC)(') exhibit good correlations with cos(2) theta;(2) combined with an exponential dependence on r(HO)('). The correlation is further improved by including a dependence on the dihedral angle rho. For each of the 34 H-bonds having observable interresidue coupling in the immunoglobulin binding domain of streptococcal protein G, a formamide dimer was generated from the crystallographic structure with energy optimized donor H-atom positions. The computed coupling constants are in reasonable agreement with the experimental, and there are excellent correlations with the simple equations involving theta;(2) and r(HO) if alpha-helix and beta sheet regions are treated separately. This dichotomy is removed by introducing the dependence on the dihedral angle rho. Justification for the use of formamide dimers is provided by almost identical interresidue coupling constants for larger sequences extracted from the X-ray structure. The amide donor (1)H chemical shifts, which were based on DFT and GIAO (gauge including atomic orbital) methods, are in poorer agreement with the experimental data but exhibit excellent correlation with r(HO)('), theta(2), and rho. PMID- 11942856 TI - Perturbation of free oligosaccharide trafficking in endoplasmic reticulum glucosidase I-deficient and castanospermine-treated cells. AB - Free oligosaccharides (FOS) are generated both in the endoplasmic reticulum (ER) and in the cytosol during glycoprotein biosynthesis. ER lumenal FOS possessing the di-N-acetylchitobiose moiety at their reducing termini (FOSGN2) are exported into the cytosol where they, along with their cytosolically generated counterparts possessing a single N-acetylglucosamine residue at their reducing termini (FOSGN1), are trimmed in order to be imported into lysosomes for final degradation. Both the ER and lysosomal FOS transport processes are unable to translocate triglucosylated FOS across membranes. In the present study, we have examined FOS trafficking in HepG2 cells treated with the glucosidase inhibitor castanospermine. We have shown that triglucosylated FOSGN2 generated in the ER are transported to the Golgi apparatus where they are deglucosylated by endomannosidase and acquire complex, sialic acid-containing structures before being secreted into the extracellular space by a Brefeldin A-sensitive pathway. FOSGN2 are also secreted from glucosidase I-deficient Lec23 cells and from the castanospermine-treated parental Chinese-hamster ovary cell line. Despite the secretion of FOSGN2 from Lec23 cells, we noted a transient intracellular accumulation (60 nmol/g cells) of triglucosylated FOSGN1 in these cells. Finally, in glucosidase I-compromised cells, FOS trafficking was severely perturbed leading to both the secretion of FOSGN2 into the extracellular space and a growth dependent pile up of triglucosylated FOSGN1 in the cytosol. The possibility that these abnormalities contributed to the severe and rapidly progressive pathology in a patient with congenital disorders of glycosylation type IIb (glucosidase I deficiency) is discussed. PMID- 11942858 TI - Perioperative medicine - a new sub-speciality, or a multi-disciplinary strategy to improve perioperative management and outcome? PMID- 11942857 TI - Insulin regulation of hepatic insulin-like growth factor-binding protein-1 (IGFBP 1) gene expression and mammalian target of rapamycin (mTOR) signalling is impaired by the presence of hydrogen peroxide. AB - Hepatic expression of insulin-like growth factor-binding protein-1 (IGFBP-1) is rapidly and completely inhibited by insulin. The signalling pathway that mediates this effect of insulin requires the activation of phosphoinositide 3-kinase (PI 3 kinase). Many of the cellular actions of insulin, including activation of PI 3 kinase, can be 'mimicked' by oxidative stresses, such as H(2)O(2). In the present study, we demonstrate that H(2)O(2) does not 'mimic' but rather antagonizes insulin repression of IGFBP-1 gene expression in H4IIE cells. This effect is accompanied by a decrease in the insulin-induced activation of mammalian target of rapamycin (mTOR)-dependent signalling. However, insulin-induced phosphorylation and regulation of protein kinase B, glycogen synthase kinase-3 and FKHR (forkhead in rhabdomyosarcoma) are not affected by H(2)O(2) in the same cells. In addition, H(2)O(2) strongly activates the p42/p44 mitogen-activated protein kinases, but the presence of PD184352 (an inhibitor of this pathway) does not block the effect of H(2)O(2) on IGFBP-1 gene expression. Our results support the view that the insulin-mediated repression of IGFBP-1 gene expression is partly mTOR-dependent, and demonstrate that H(2)O(2) selectively antagonizes mTOR dependent insulin action. The implications for the use of H(2)O(2)-generating agents as therapeutics for the treatment of insulin resistance, as well as the role of oxidative stress in the development of insulin resistance, are discussed. PMID- 11942859 TI - Preconditioning - endogenous defence mechanisms of the heart. AB - The term 'preconditioning' refers to the paradoxical phenomenon that pretreatment with a potential noxious stress-stimulus can increase cellular tolerance to subsequent noxious stress-stimuli. This was first described in an experimental model in dogs in which short-lasting periods of myocardial ischemia resulted in reduced infarction during a subsequent long-lasting coronary artery occlusion. Similar observations have also been made in other species and in other organs. During the last few years, the term preconditioning has been expanded to include pretreatment with other physical stress-stimuli or pharmacological agents that can increase cellular resistance to injury. The phenomenon probably represents a general adaptive response to cellular stress, but mechanisms involved are not fully clarified. This review focuses on preconditioning in the heart. Firstly, we want to address the observation that activation of endogenous defence mechanisms can increase cellular tolerance to several potentially noxious stimuli. Based on results from experimental research, we will give an overview of intracellular mechanisms that is currently in focus. Secondly, we want to address the potential role of preconditioning in clinical practice. We will present results from studies in patients with coronary artery disease and discuss possible clinical implications. Results show that the phenomenon probably exists in the human myocardium. In the future, this might be exploited in patients with acute coronary syndromes, especially since advanced techniques are now available for acute revascularization. Additionally, identification of possible mechanisms involved may influence the choice of medical treatment in high-risk patients with stable coronary artery disease. Preconditioning can also be exploited during elective surgical procedures. This should be of great interest, as the extent of elective surgery in patients at high-risk for coronary events is increasing. In this respect it is important to note that opioid-receptors are probably involved in preconditioning in humans. The last part of this review will address the possible relation between preconditioning and different anesthetic agents and sedatives. PMID- 11942860 TI - Effect of hemodiafiltration and sepsis on chemotaxis of granulocytes and the release of IL-8 and IL-10. AB - BACKGROUND: Extracorporeal circulation, such as cardiopulmonary bypass and hemodialysis, has been associated with an activation of the immune system. Continuous veno venous hemodiafiltration (CVVHD) is used in critically ill septic patients. During CVVHD, cytokines are excreted in ultrafiltrate. When the membranes, used in CVVHD, are incubated with leukocytes in vitro a slight production of cytokines is observed. Due to the underlying disease it is difficult to investigate the effect of CVVHD in septic patients. We therefore studied the separate effect of CVVHD on the chemotaxis of granulocytes, the proliferation of lymphocytes and the release of IL-8 and IL-10 in healthy pigs compared to an endotoxin and a control group. METHODS: Thirty-one pigs were anesthetized and mechanically ventilated. CVVHD was performed in 10 pigs. Eleven pigs received an infusion of Escherichia coli endotoxin 30 microg/kg, and 10 pigs served as a control group. The chemotaxis of granulocytes was measured in an assay chamber, and the cytokines IL-8 and IL-10 with an enzyme-linked immunosorbent assay. The adhesion molecules CD18 and CD62 on lymphocytes were measured using monoclonal antibodies, and the lymphocyte proliferation was measured without stimulation and in response to mitogens. RESULTS: CVVHD was accompanied by lymphocytopenia and increased spontaneous lymphoproliferative response, but no change in adhesion molecules on lymphocytes or cytokine levels in plasma, and no decrease in the chemotaxis of granulocytes. Following endotoxin we observed a pronounced lymphocytopenia and an increased secretion of IL-8 and IL-10, a decrease in the expression of CD18 on lymphocytes and in the stimulated lymphocyte proliferation and in the chemotaxis of granulocytes. CONCLUSION: CVVHD does not, in contrast to endotoxin-induced sepsis, influence chemotaxis of granulocytes, the production of IL-8 and IL-10 or the proliferation of lymphocytes. PMID- 11942861 TI - Adrenocortical function and multiple organ failure in severe sepsis. AB - BACKGROUND: Some patients with severe sepsis may have relative adrenocortical insufficiency, although not all studies confirm this finding. Corticosteroids play an important role in controlling excessive immune response, and they may reduce the severity of organ dysfunction in critical illness. In this prospective study, we investigated the incidence of adrenal insufficiency in severe sepsis and its relation to the development of multiple organ failure. METHODS: Forty-one patients meeting the criteria for severe sepsis were studied. A short ACTH stimulation test was carried out within 24 h of the diagnosis of sepsis. Peak serum cortisol level < 680 nmol/L and a rise of less than 260 nmol/L were used as the criteria for relative adrenocortical insufficiency. RESULTS: Relative adrenocortical insufficiency was detected in six patients. Duration of the ICU stay (P = 0.002) and mechanical ventilation (P = 0.024) were significantly longer in patients with impaired adrenal function. In the survivors, SOFA scores were significantly higher in patients with impaired adrenal function. The plasma ACTH levels were normal in most of the patients with relative adrenal insufficiency, whereas most patients with normal adrenal function had extremely low plasma ACTH levels. CONCLUSION: The ICU stay was longer and multiple organ failure more severe in patients with impaired adrenocortical function. There was a clear dissociation between ACTH and cortisol levels in AAR patients. This finding suggests that the integrity of the hypothalamic-pituitary-adrenal axis may be impaired in severe sepsis. PMID- 11942862 TI - Clinical evaluation of a partial CO2 rebreathing technique for cardiac output monitoring in critically ill patients. AB - BACKGROUND: Monitoring central hemodynamics is essential in critically ill patients and less invasive techniques are needed. In this study, the clinical and technical performance of a new non-invasive cardiac output monitor (NICO) based on partial CO2 rebreathing technique and a modified Fick equation were evaluated. The various sources of possible errors in measurement of cardiac output (CO), carbon dioxide production (VCO2) and pulmonary shunt were also assessed. METHODS: Simultaneous measurements of CO with partial CO2 rebreathing technique (CO(nico)) and thermodilution (CO(td)) were performed in 15 patients during major surgery or in the ICU. Pulmonary shunt was estimated from this device and compared to values obtained by standard shunt formula. The accuracy of VCO2 measurements was assessed in a mechanical lung model. RESULTS: A good correlation was found between CO(nico) and CO(td) (r = 0.96, within-subject correlation r = 0.88) with a small underestimation of cardiac output by the NICO of 0.04 L/min, limits of agreement (+/- 2 SD) being - 1.68 and 1.76 L/min. In hemodynamic unstable patients the method closely tracked changes in CO. Pulmonary shunt was underestimated by approximately 11%-units compared to standard shunt calculations using arterial and mixed venous blood gases. We also observed an underestimation in VCO2 measurements. CONCLUSION: Clinical evaluation shows that partial CO2 rebreathing technique provides a useful and accurate non-invasive estimate of cardiac output. Although this technique cannot fully replace the pulmonary artery catheter, it may be used to monitor central hemodynamics in a large number of critically ill patients. PMID- 11942863 TI - Sympathetic nervous activation following subarachnoid hemorrhage: Influence of intravenous clonidine. AB - BACKGROUND: Subarachnoid hemorrhage is often accompanied by systemic complications and cerebral vasospasm. Elevated levels of circulating catecholamines may be involved in the pathophysiology behind these events. The alpha-2-agonist clonidine inhibits sympathetic outflow by a central mechanism. Unrestricted sympathoexcitation may be detrimental and administration of clonidine may be beneficial in these patients. METHODS: Using isotope dilution methodology, norepinephrine kinetic determinations, comprising determination of arterial norepinephrine concentration and rates of norepinephrine spillover to and removal, or clearance, from plasma, were performed on three occasions during the first week after subarachnoid hemorrhage in 25 patients. Eleven of these patients received clonidine (continuous i.v. infusion 5.8 +/- 0.7 microg x kg(-1) x 24 h(-1)) and the remainder, standard therapy. Initial results were compared with 17 healthy age-matched subjects and eight patients suffering from severe traumatic brain injury without traumatic subarachnoid hemorrhage. RESULTS: Subarachnoid hemorrhage patients exhibited markedly elevated arterial plasma norepinephrine concentrations [3.74 +/- 0.48, P < 0.001 vs. healthy subjects (1.59 +/- 0.11 nmol/L) and P < 0.05 vs. head trauma patients (1.94 +/- 0.29 nmol/L)]. The rate of clearance of norepinephrine from plasma in the subarachnoid patients was also significantly greater than that observed in the healthy subjects (2.66 +/- 0.15 vs. 2.14 +/- 0.15 L/min, P < 0.05) and the head trauma patients (2.00 +/- 0.12 L/min, P < 0.05). Compared with both control groups, on admission the rate of spillover of norepinephrine to plasma following subarachnoid hemorrhage was markedly elevated (9.11 +/- 1.12, P < 0.001). Clonidine treatment (continuous i.v. infusion 5.8 +/- 0.7 microg x kg(-1) x 24 h( 1)) did not reduce the increased rate of spillover of norepinephrine to plasma following subarachnoid hemorrhage. CONCLUSION: Sympathetic nervous activity is markedly elevated following subarachnoid bleeding. Clonidine had no effect on the rate of norepinephrine spillover to, or clearance from, plasma in these patients. Clearly, further studies are required to elucidate the mechanisms responsible for generating sympathetic nervous activation following subarachnoid hemorrhage. PMID- 11942864 TI - Albumin extravasation and tissue washout of hyaluronan after plasma volume expansion with crystalloid or hypooncotic colloid solutions. AB - BACKGROUND: Intravascular volume expansion is followed by loss of fluid from the circulation. The extravasation of albumin in this readjustment is insufficiently known. METHODS: Twelve male volunteers participated, each in three separate sessions, in a controlled, randomised, open fashion. They received one of the following: albumin 40 g/L,(7.1 mL/kg, i.e. 500 mL per 70 kg); Ringer's acetate (21.4 mL/kg), or dextran 30 g/L (7.1 mL/kg). The fluids were infused during 30 min and the subjects were followed for 180 min. ECG, arterial oxygen saturation and non-invasive arterial pressure were recorded. Haemoglobin, haematocrit, serum albumin and osmolality, plasma colloid osmotic pressure and hyaluronan concentration were determined in venous samples. RESULTS: The serum albumin concentration decreased (P < 0.05, anova) following Ringer's acetate or dextran, whereas serum osmolality was unchanged in all groups. The colloid osmotic pressure decreased (P < 0.05) after the Ringer solution. The blood volume increase was estimated from the decrease in haemoglobin concentration and did not differ between the three fluids. The cumulated extravasation of albumin was largest following albumin (10.4 +/- 5.4 g, mean +/- SD), less following dextran (5.6 +/- 5.0 g) and negligible in the Ringer group (0.5 +/- 10.0 g; P < 0.05 against albumin). However, the Ringer solution increased the plasma concentration of hyaluronan drastically. CONCLUSIONS: Infusion of hypotonic colloidal solutions entails net loss of albumin from the vascular space. This is not the case after Ringer's acetate. Increased interstitial hydration from the latter fluid is followed by lymphatic wash out of hyaluronan. PMID- 11942865 TI - Accuracy of radial arterial pressure measurement during surgery under controlled hypotension. AB - BACKGROUND: Radial arterial pressure underestimates the pressure in the aorta in several clinical situations. A central-to-radial pressure gradient was attributed to intense vasodilation. The aim of this study was to evaluate the accuracy of radial pressure monitoring during controlled hypotension achieved with profound arterial vasodilation. METHODS: Ten patients with ASA physical status I and II undergoing maxillofacial surgery under general anesthesia were enrolled in this prospective study. Radial and femoral arteries were cannulated and connected to a pressure monitoring system. Controlled hypotension was achieved with an infusion of nicardipine titrated to maintain MAP between 50 and 60 mmHg. Simultaneous radial and femoral systolic, mean and diastolic arterial pressures were recorded before, during and after controlled hypotension. Results were expressed as mean +/- SD. Concomitant radial and femoral pressures were compared by a paired Student's test, P < 0.05 being significant. RESULTS: In all, 150 sets of arterial pressures measurement were obtained. There were no statistically significant differences between radial and femoral arterial pressures measured before, during or after controlled hypotension. CONCLUSION: Radial arterial pressure is an accurate measure of central arterial pressure during controlled hypotension achieved with arterial vasodilation. PMID- 11942866 TI - Effects of intravenous anesthetics on interleukin (IL)-6 and IL-10 production by lipopolysaccharide-stimulated mononuclear cells from healthy volunteers. AB - BACKGROUND: Surgical trauma has been shown to augment the plasma concentrations of proinflammatory cytokines, which are important mediators of host defense mechanisms and the systemic inflammatory response syndrome (SIRS). Recently, it has been shown that certain kinds of surgery provoke not only a proinflammatory response (SIRS) but also a concurrent anti-inflammatory response. The aim of this study was therefore to examine the effects of intravenous anesthetics on the synthesis of interleukin (IL)-6 (a proinflammatory cytokine) and IL-10 (an anti inflammatory cytokine) by lipopolysaccharide (LPS)-stimulated mononuclear cells from healthy volunteers. METHODS: Peripheral blood mononuclear cells (PBMCs) from 17 healthy volunteers, separated by centrifugation on a Ficoll-Hypaque gradient, were washed and suspended in RPMI containing 10% heat-inactivated fetal calf serum (FCS). After adding RPMI-FCS containing various concentrations of intravenous anesthetics (propofol, thiopental, ketamine and midazolam), the PBMCs were incubated overnight in the presence of a submaximal concentration of LPS. The supernatants were collected and their IL-6 and IL-10 contents were assayed using enzyme-linked immunosorbent assay kits. RESULTS: Propofol inhibited both IL 6 and IL-10 production at 0.5 microg/mL, 5 microg/mL and 50 microg/mL. Conversely, thiopental induced IL-10 production at 2 microg/mL and 20 microg/mL. CONCLUSION: Propofol appears to inhibit both IL-6 and IL-10 production by LPS stimulated PBMCs in vitro. Further study is required to clarify the mechanism of the suppressive effect of propofol. PMID- 11942867 TI - Carbon monoxide poisoning - a cause of increased QT dispersion. AB - BACKGROUND: Carbon monoxide (CO) poisoning is associated with direct cardiovascular toxicity. QT dispersion (QTd) of the ECG is an indirect measure of heterogeneity of ventricular repolarization, which may contribute to ventricular arrhythmias. Our aim was to study QTd in patients with acute CO poisoning. METHODS: CO intoxication was confirmed by arterial blood gas analysis. A control group consisted of age- and sex-matched individuals admitted to the hospital for unrelated clinical conditions. 12-lead ECG's were recorded on admission and repeated 1 week after discharge from the hospital. QT dispersion was defined as the difference between the greatest and the least QT intervals in any of the 12 leads. RESULTS: Seventeen intoxicated patients, aged 5-46 years, had mean carboxyhemoglobin levels of 22.5 +/- 11.1%. On admission, corrected QT intervals of the intoxicated patients were significantly increased compared to the control group (431 +/- 18 ms vs. 404 +/- 28 ms, P = 0.008), but not the QT interval (358 +/- 25 ms vs. 345 +/- 20 ms, P = 0.17). Mean QTd and cQTd values (46 +/- 15 ms and 62 +/- 13 ms) of the intoxicated patients were significantly increased compared to the control group (17 +/- 4 ms and 33 +/- 15 ms, P < 0.0001 for both). Both QTd and cQTd decreased significantly after discharge from the hospital (P = 0.0001). CONCLUSION: Although QT dispersion increased in patients with CO poisoning, none of ECG's showed ventricular arrhythmia. Increased QTd in the absence of QT interval prolongation may have a lowered arrhythmogenic potential of CO poisoning. PMID- 11942868 TI - Parenteral ketoprofen for pain management after adenoidectomy: comparison of intravenous and intramuscular routes of administration. AB - BACKGROUND: Different parenteral routes of administration of NSAIDs such as ketoprofen have not been properly compared in children. This study was designed to compare the analgesic efficacy of intravenous and intramuscular ketoprofen for pain management in children after day-case adenoidectomy. METHODS: A total of 120 children, aged 1-9 years, who were scheduled to undergo adenoidectomy, were randomized to receive ketoprofen 2 mg/kg either intravenously with intramuscular placebo (n = 40) or ketoprofen 2 mg/kg intramuscularly with intravenous placebo (n = 40), or both intravenous and intramuscular placebo (n = 40) at induction of anesthesia. The study design was prospective and double-blind with parallel groups. Pain was assessed at rest and during swallowing using the Maunuksela pain scale during 3 h after surgery, and fentanyl i.v. was given for rescue analgesia. RESULTS: Children in the Placebo group needed significantly more doses of fentanyl (72 doses) than either children in the intravenous group (47 doses) or children in the intramuscular group (51 doses) (P = 0.021). In addition, a higher proportion of children in the Placebo group than in the two ketoprofen groups (P = 0.03) demanded rescue analgesic. No difference in the need for rescue analgesia or in pain scores was found between the two ketoprofen groups. Children in the intravenous group had less pain than children in the Placebo group. The difference was significant during swallowing at 1 h after surgery (P = 0.046) and for the worst pain observed during swallowing for 3 h after surgery (P = 0.022). There were no differences between the three groups with respect to operation times, amount of perioperative bleeding, or rate or extent of adverse events. CONCLUSION: The efficacy of intravenous and intramuscular ketoprofen was similar, and they both differed from placebo. PMID- 11942869 TI - Analgesia for pelvic and perineal cancer pain by intrathecal steroid injection. AB - BACKGROUND: Corticosteroids are used systemically in patients with advanced cancer to alleviate pain. Intrathecal administration of steroids is rarely performed but analgesic effects of intrathecal steroids have been reported in animal studies. We administered betamethasone intrathecally in three cancer patients with uncontrollable pain. CASE REPORT: Intrathecal injection of betamethasone (1-4 mg) with saline (total volume = 2 mL) was performed in three patients with advanced pelvic or perineal cancer, in whom pain could not be controlled in spite of various analgesic therapies. After obtaining the patient's informed consent for the procedure, betamethasone was administered intrathecally through the L4/5 intervertebral space. Intrathecal betamethasone produced rapid analgesia within 10 min and subsequent long-lasting analgesia for 5 days or more. Sleep, appetite and activity improved. No adverse effects were observed in any of the patients. CONCLUSIONS: Intrathecal injection of betamethasone may be a useful approach in some patients with intractable cancer pain. PMID- 11942870 TI - Pharmacokinetics of a 24-hour intravenous ketoprofen infusion in children. AB - BACKGROUND: No pharmacokinetic data are available with respect to the plasma concentration of ketoprofen during intravenous infusion in children. METHODS: We present here the pharmacokinetics of ketoprofen after a 10-min intravenous infusion of 1 mg/kg followed by a 24-h infusion of 4 mg/kg in 18 children aged 7 months to 16 years. Venous blood samples were collected at 5 min, 1, 2, 4, 24 h following the loading dose, and then 1, 2 and 4 h after the end of the infusion. A validated HPLC method was used to measure plasma levels of ketoprofen. RESULTS: The steady state plasma concentration of ketoprofen was 2.0 microg/mL (range 1.3 2.7 microg/mL). The clearance of ketoprofen was 0.09 L x h(-1) x kg(-1) (range 0.06-0.13 L x h(-1) x kg(-1)). The distribution volume was 0.16 L/kg (range 0.12 0.21 L/kg). The terminal half-life was 1.3 h (range 0.8-1.7 h). CONCLUSION: The pharmacokinetics of ketopofen in children is similar to that reported in adults. Our results indicate that ketoprofen is a feasible drug for continuous intravenous infusion in acute pain treatment in children. PMID- 11942871 TI - Anatomical study applied to anesthetic block technique of the superior laryngeal nerve. AB - BACKGROUND: The topography of the internal branch of the superior laryngeal nerve (ibSLN) was prospectively studied to evaluate the greater horn of the hyoid bone (ghHB) and the incisura of the thyroid cartilage (iTC) as anatomical repairs in laryngeal anesthetic block. Factors such as gender, ethnicity and side of the neck were also analyzed concerning their influence in the ibSLN position. METHODS: One hundred neck dissections were performed in 50 human cadavers bilaterally identifying the ibSLN, the ghHB and iTC. The distance between the ghHB and ibSLN in the cranio-caudal direction (dHB), and the distance between the iTC and the ipsilateral thyrohyoid membrane ostium (dTC) were measured. Furthermore, the results were statistically analyzed according to ethnicity, gender and side of the neck. RESULTS: The ibSLN was juxtaposed to the apex ghBH in 31 out of 100 dissections. The mean dHB was 2.4 mm, and mean dTC was 33.4 mm. The statistical analysis did not identify any significant difference regarding those distances between the groups in terms of ethnicity, gender and side of the neck. CONCLUSION: The ibSLN was often dissected very close to the ghHB, and this result was not influenced by any factor studied. Therefore, the ghHB can be considered a good anatomical repair to localize the ibSLN in the local block of the larynx. Furthermore, the dTC could frequently be reached by routinely used nerve block needle. However, a few anatomical variations may occur, resulting in a low failure rate of this anesthetic procedure. PMID- 11942872 TI - EMLA cream prior to digital nerve block for ingrown nail surgery does not reduce pain at injection of anesthetic solution. AB - BACKGROUND: Needle penetration and local anesthetic infiltration are painful steps of digital ring block. The objective of this study was to evaluate the efficacy of EMLA cream application prior to digital ring block for surgery for ingrown big toenail. METHODS: We conducted a prospective, double-blinded, placebo controlled, randomized clinical trial with 81 patients (range: 12-75 years, mean +/- SD: 30.3 +/- 17.5 years), who underwent big toenail surgery. Prior to the digital block, EMLA cream (Group E) or a placebo cream (Group P) was applied. A visual analog scale (VAS) and a verbal rating score (VRS) from 1 to 10, 10 being most severe pain, were used for assessment of pain during the skin needle penetration and during the infiltration of the anesthetic product. RESULTS: Data of the VAS and of the VRS during the two steps, respectively, show no significant difference in pain rating between the two groups. In Group E, the gender of the patients had a significant effect on pain perception. Male patients reported less pain than female patients, both during the needle skin penetration and during the infiltration step (P < 0.005). In Group P, reported pain did not differ by gender in either step of the procedure. CONCLUSIONS: We found no clinical benefit in using EMLA during digital nerve block. Although the digital nerves are not deeply located in the small subcutaneous space of the digit, topical anesthesia may not reach them. Moreover, swelling due to injection of the anesthetic product in this small space may also amplify the sensation of pain. PMID- 11942873 TI - Postoperative muscle paralysis after rocuronium: less residual block when acceleromyography is used. AB - BACKGROUND: Residual muscle paralysis after anesthesia is common after pancuronium, but less common following the intermediate-acting drugs vecuronium and atracurium. Therefore, many anesthetists do not monitor neuromuscular function when using an intermediate-acting agent. The purpose of this prospective, randomised and double-blind study was to establish the incidence and degree of postoperative residual block following the use of rocuronium in patients not monitored with a nerve stimulator, and to compare it with results obtained in patients monitored using acceleromyography (AMG). METHODS: During propofol/opioid anesthesia, 120 adult patients were randomised to two groups, one monitored with AMG, the other using only clinical criteria without a nerve stimulator. Postoperatively, TOF-ratio was measured with mechanomyography; a TOF ratio < 0.80 indicated residual muscle paralysis. RESULTS: Residual muscle paralysis was found in 10 patients in the group without neuromuscular monitoring (16.7%) (95% confidence interval, 12-21%) and in two patients in the AMG monitored group (3%) (95% CI, 0-8%); (P = 0.029, Fisher's exact test). Time from end of surgery to tracheal extubation was significantly longer in the AMG monitored group (12.5 min) than in the group not monitored with AMG (10 min). CONCLUSION: Clinical evaluation of recovery of neuromuscular function does not exclude significant residual paralysis following the intermediate-acting muscle relaxant rocuronium, but the problem of residual block can be minimized by use of AMG. PMID- 11942874 TI - Effect of metoclopramide on mivacurium-induced neuromuscular block. AB - In order to investigate the effect of metoclopramide on the duration of action of mivacurium, 45 patients were randomized into three groups. Group M10 (n = 15) and M20 (n = 15) received 10 and 20 mg of metoclopramide i.v., respectively, and group S (n = 15) received saline 2 min before induction of anesthesia with fentanyl, thiopental and mivacurium. Plasma cholinesterase activity (pCHE) was measured before induction of anesthesia and 2 min after injection of metoclopramide and saline. Neuromuscular block was monitored by a force transducer using train of four nerve stimulation. Anesthesia was maintained with isoflurane and N2O. Time to recovery of a twitch height of 90% was significantly prolonged in group M10 and M20 (44 +/- 15 and 57 +/- 10 min) as compared to group S, 32 +/- 9 min, P < 0.05). A slight but significant decrease in pCHE was observed in group M20. Because of the risk of prolonged duration of action of mivacurium, neuromuscular blockade should always be monitored whenever metoclopramide is given before injection of mivacurium. PMID- 11942875 TI - Postoperative epidural hematoma or cerebrovascular accident? A dilemma in differential diagnosis. AB - An elderly lady developed an epidural hematoma following combined spinal-epidural anesthesia with a local anesthetic-opioid mixture for a vaginal hysterectomy. This occurred in association with the use of prophylactic subcutaneously administered unfractionated heparin. She had diabetes, hypertension and had previously undergone coronary artery bypass surgery and right carotid endarterectomy. Warfarin and aspirin were discontinued 2 weeks before the surgery. Postoperatively, an atypical presentation of backache, bilateral sensory loss and left lower limb monoplegia ensued. The initial clinical impression was of a cerebrovascular accident. Magnetic resonance imaging, however, revealed an extensive epidural hematoma that necessitated decompression laminectomy. Progression to paraparesis occurred but the patient gradually regained much of her functionality over the next 2 years. PMID- 11942876 TI - Perioperative management of long QT syndrome in a child with congenital heart disease. AB - During cardiac catheterization, a 2(1/2)-year-old boy developed sudden cardiac arrest. The presence of a long QT interval in the electrocardiogram (ECG) along with ventricular arrhythmia and syncope at that moment enabled us to diagnose long QT syndrome (LQTS). Immediate defibrillation and beta-blocker (metoprolol) therapy saved the life of the child. Cardiac catheterization was completed and the child was planned for Fontan operation. Beta-blocker coverage, prevention of sympathetic stimulation and avoidance of agents which prolong the QT interval made anesthesia uneventful. There were episodes of ventricular fibrillation (VF) in the postoperative period. The child was managed with electrical defibrillation, metoprolol and magnesium. PMID- 11942877 TI - Central anticholinergic syndrome in a child undergoing circumcision. AB - We describe one of the few pediatric cases of central anticholinergic syndrome (CAS) in an 8-year-old boy undergoing elective surgery. Deep sedation, inadequate response to stimuli and reduced muscular tone of the upper airway resulting in airway obstruction were the clinical manifestations of CAS. The symptoms resolved immediately after administration of physostigmine. This case illustrates the importance of considering central anticholinergic syndrome as a differential diagnosis in children if prolonged sedation after general anesthesia occurs. PMID- 11942878 TI - Subcutaneous emphysema and respiratory failure managed with infraclavicular blow holes. PMID- 11942879 TI - Condoms in the age of AIDS. PMID- 11942880 TI - Prolactin receptor in human endometriotic tissues. AB - BACKGROUND: Augmented serum prolactin (PRL) levels have been implicated as a cause for infertility in patients with endometriosis. However, it has not been established whether this lactogenic hormone or its receptor (PRLR) have specific effects on the development of human endometriosis. In the present study we assessed PRLR expression in human endometriotic and normal tissue. METHODS: Fourteen patients with laparoscopically proven endometriosis were recruited and the negative or positive expression of the PRLR in normal endometrial and endometriotic tissues obtained during the mid-late proliferative phase of the menstrual cycle was evaluated by RT-PCR. Evaluation of serum prolactin, and peritoneal fluid estradiol content were carried out by specific radioimmunoassays (RIA). RESULTS: In endometriotic tissue samples the corresponding 1048 bp PRLR transcript was negative in 12/14 of the samples (86%) and positive in two samples (14%). The opposite was seen in normal tissue, since PRLR expression was positive in 11/14 samples (79%) and negative in the remaining three samples (21%). There was an association between serum prolactin concentrations and PRLR expression in normal tissue but not in endometriotic tissue. CONCLUSIONS: The PRLR is expressed in normal endometrium, while cells of the endometriotic tissues did not express the PRLR, at least during the mid-late proliferative phase of the menstrual cycle. The absence of the PRLR in the endometriotic tissues could not be correlated with the serum prolactin levels. These results suggest the existence of differential regulation of PRLR expression between normal and endometriotic tissue. PMID- 11942881 TI - Predictive factors of developing diabetes mellitus in women with gestational diabetes. AB - BACKGROUND: To investigate which factors during gestational diabetes pregnancies correlate with the risk of developing impaired glucose tolerance or diabetes 1 year postpartum and to compare this risk in women with gestational diabetes and women with a normal oral glucose tolerance test during pregnancy. METHODS: Of 315 women with gestational diabetes, defined as a 2-hr blood glucose value of at least 9.0 mmol/l at a 75-g oral glucose tolerance test, who delivered in Lund 1991-99, 229 (73%) performed a new test 1 year postpartum. We compared maternal and fetal factors during pregnancy with the test value at follow up. A control group of 153 women with a 2-hr test value below 7.8 mmol/l during pregnancy were invited to a new test 1 year postpartum and 60 (39%) accepted. RESULTS: At 1 year follow up, 31% of the women with gestational diabetes but only one of the 60 controls showed pathologic glucose tolerance and one had developed diabetes. The following factors in women with gestational diabetes were identified as predicting impaired glucose tolerance or diabetes at 1 year follow up: maternal age over 40 and--in a multiple regression analysis, independent of each other--a high 2-hr value at oral glucose tolerance test during pregnancy and insulin treatment during pregnancy. CONCLUSION: The risk of developing manifest diabetes after gestational diabetes may be high enough to justify a general screening or diagnostic procedure in all pregnant women to identify women with gestational diabetes and a postpartum follow up program for them. This study did not identify any particular factor during pregnancy with enough precision to predict a later progression to diabetes. PMID- 11942882 TI - Substandard factors in perinatal care in The Netherlands: a regional audit of perinatal deaths. AB - BACKGROUND: To determine: 1) whether substandard factors were present in cases of perinatal death, and to what extent another course of action might have resulted in a better outcome, and 2) whether there were differences in the frequency of substandard factors by level of care, particularly between midwives and gynecologists/obstetricians and between home and hospital births. METHODS: Population-based perinatal audit, with explicit evidence-based audit criteria. SETTING: The northern part of the province of South-Holland in The Netherlands. All levels of perinatal care (primary, secondary and tertiary care, and home and hospital births) were included. CASES: Three hundred and forty-two cases of perinatal mortality (24 weeks of pregnancy--28 days after birth). MAIN OUTCOME MEASURES: Scores by a Dutch and a European audit panel. Score 0: no substandard factors identified; score 1, 2 or 3: one or more substandard factors identified, which were unlikely (1), possibly (2) or probably (3) related to the perinatal death. RESULTS: In 25% of the perinatal deaths (95% Confidence Interval: 20-30%) a substandard factor was identified that according to the Dutch panel was possibly or probably related to the perinatal death. These were mainly maternal/social factors (10% of all perinatal deaths; most frequent substandard factor: smoking during pregnancy), and antenatal care factors (10% of all perinatal deaths; most frequent substandard factor: detection of intra-uterine growth retardation). We did not find statistically significant differences in scores between midwives and gynecologists/obstetricians or between home and hospital births. The European panel identified more substandard factors, but these were again equally distributed by level of care. CONCLUSIONS: Perinatal deaths might be partly preventable in The Netherlands. There is no evidence that the frequency of substandard factors is related to specific aspects of the perinatal care system in The Netherlands. PMID- 11942883 TI - Nifedipine or hydralazine as a first-line agent to control hypertension in severe preeclampsia. AB - BACKGROUND: Pre-eclampsia is one of the most serious and common complications of pregnancy. Nifedipine, a calcium channel blocker, and the vasodilator hydralazine have both been used as antihypertensive agents in this condition. The aim of this study was to determine which of these two agents is the most appropriate antihypertensive in the management of severe pre-eclampsia. METHODS: One hundred and twenty-six pre-eclamptic patients with a gestational age of more than 20 weeks were randomized to receive either 8 mg nifedipine sublingually or 5-10 mg intravenous hydralazine. Women with a history of heart failure and women receiving antihypertensive treatment during the course of the current pregnancy were excluded. For each patient the following data were recorded; the number of drug administrations, the time needed to control blood pressure, mean urinary output, the time interval between effective control and a new hypertensive crisis after each drug administration and relevant adverse effects in mother or fetus. RESULTS: Effective control of blood pressure was achieved in both treatment arms. Data analysis indicated significantly fewer drug administrations in the nifedipine arm of the study. The time interval before a new hypertensive crisis following initial effective control of blood pressure was significantly longer in the nifedipine group when compared with hydralazine. Effective control of blood pressure was achieved more rapidly in multiparous patients receiving nifedipine (p=0.026). Mean urinary output before and after delivery was greater in the nifedipine arm of the study. There were no significant differences between the two groups in other variables. In addition, in neither group were there any serious adverse effects in mother or fetus. CONCLUSION: Nifedipine is safe and more effective than hydralazine in controlling blood pressure in severe pre eclampsia. It has the added advantage of being cheaper and more widely available than the latter and is easily administered. PMID- 11942884 TI - Normal cervical changes in parous women during the second half of pregnancy--a prospective, longitudinal ultrasound study. AB - OBJECTIVE: To determine what constitutes normal cervical changes during the second half of pregnancy in parous women delivering at term. DESIGN: The study comprises 21 healthy, pregnant parous women who all gave birth at term. They were examined with transvaginal ultrasound every two weeks from 24 gestational weeks until delivery. Cervical length and width were measured. The inner cervical os was assessed as being closed or open, the length and width of any opening were measured, and dynamic cervical changes (i.e. opening and closing of the inner cervical os during examination) were noted. RESULTS: Median cervical length was 41 mm (range 26-55) at the first examination and 29 mm (range 8-56) at the last examination. The corresponding figures for cervical width were 38 mm (range 29 47) and 46 mm (range 38-64). Cervical length decreased in 18 women but remained unchanged in three. Three patterns of change in cervical length were observed: in 12 women there was a steady, continuous decrease in cervical length (median decrease rate 1.1 mm/week, range 0.6-2.4); in four women the decrease rate accelerated towards the end of pregnancy, the median decrease rate after the change being 3.0 mm/week (range 1.5-4.8); and in two women there was a sudden drop in cervical length at term. Cervical width increased in 16 women but remained unchanged in five. Two patterns of change in cervical width were seen: 14 women manifested a steady continuous increase in cervical width (median 0.8 mm/week, range 0.4-1.8); in two women the increase rate accelerated from around 34 gestational weeks, the increase rate after the change being 4.1 and 5.9 mm/week, respectively. Opening of the internal cervical os was observed at least once in 11 (52%) women and was seen as early as at 24 and 25 gestational weeks in two women. The opening was always V-shaped (median length 6 mm, range 4-17; median width 7 mm, range 3-20). Dynamic changes of the internal cervical os were seen in three women (14%) at 25, 30 and 41 gestational weeks, respectively. CONCLUSION: The cervix of parous women decreases in length and increases in width from midpregnancy to term, but the pattern of change varies between individuals. Knowledge of the different patterns of normal change forms the basis of transvaginal ultrasound studies of pathological cervical changes during pregnancy. PMID- 11942885 TI - The role of sonographic assessment of cervical length in the prediction of preterm birth in primigravidae with twin gestation conceived after infertility treatment. AB - OBJECTIVE: To identify the risk factors for preterm birth in primigravidae with twin gestation and the role of transvaginal ultrasonographic assessment of the cervix. METHODS: Between January 1996 and December 1996, 54 twin pregnancies were prospectively enrolled. All women were at their first pregnancy. All women conceived following infertility treatments and all had a normal uterine cavity proven by hysterosalpingography (HSG) or hysteroscopy. Multiple logistic regression analysis was used to evaluate the association between the length of the cervix at 18-24 weeks of gestation and outcome variables, controlling for possible confounding factors. RESULTS: The mean +/- SD maternal age was 30.9 +/- 5.3 years (range 22-46), and five of them were aged 40 or more. Nine patients (20.5%) delivered prematurely, defined as spontaneous delivery at or before 34 weeks of gestation. There was no statistically significant difference between women who delivered before or after 34 weeks of gestation in regard to maternal age, body mass index (BMI), weight gain in pregnancy, smoking and work during pregnancy. The mean cervical length of patients who delivered before 34 weeks of gestation (30.1 +/- 6.1 mm) was significantly shorter than that of women who delivered after 34 weeks of gestation (42.2 +/- 6.2 mm; P < 0.001). Cervical length longer than 35 mm predicted delivery after 34 weeks of gestation with sensitivity and specificity of 88.5% and 88.9%, respectively. The positive and negative predictive values were 96.9% and 66.7%. CONCLUSION: A transvaginal ultrasonographic measurement of the cervix > 35 mm at 18-24 weeks in twin gestation can identify patients at low risk for delivery before 34 weeks. Maternal age, BMI, weights gain, smoking and work during the pregnancy did not influence the duration of the pregnancy. PMID- 11942886 TI - A prospective observational study on tears during vaginal delivery: occurrences and risk factors. AB - BACKGROUND: To ascertain the occurrence and distribution of various types of I-IV degree tears, during childbirth, and analyze risk factors for perineal II degree tears. MATERIALS AND METHODS: A total of 2883 consecutive vaginal deliveries, during 1995-97 at Sahlgrenska University Hospital in Goteborg Sweden, were included. All tears were classified according to an especially designed protocol, and risk factors for II degree tears were evaluated by use of univariate and logistic regression analysis. RESULTS: Only 6.6% of nulliparous parturients had no detectable tear as compared to 34.2% in parous women. Almost half of the women suffered from a II degree tear during birth, and a higher proportion of nulliparous (16.6%) than parous (9.4%) women had extensive perineal lacerations. In addition, nulliparous were more likely than parous parturients to be subjected to a perineotomy (18.1% versus 5.6%). Stepwise logistic regression analysis revealed that the following factors remained independently associated with II degree tear: slight perineal edema, high infant weight, excellent visualization of perineum, increasing age of the mother, excellent cooperation of the women, protracted second phase (> 60 min) and duration of second phase < 30 min. CONCLUSIONS: The majority of women (78%) undergoing childbirth had a tear and 47.1% suffered from perineal lacerations. Nulliparous women were more likely to have severe perineal lacerations or episiotomies. Similar risk factors were found for II degree tears as previously shown for III/IV degree tears. PMID- 11942887 TI - Accidental out-of-hospital deliveries: an obstetric and neonatal case control study. AB - BACKGROUND: Accidental out-of-hospital (OHD) deliveries are associated with high rates of perinatal morbidity and mortality. The ability of health care workers to identify women at risk of out-of-hospital delivery is limited. The purpose of this study was to determine the prevalence of these deliveries in our population and to quantify the neonatal morbidity and mortality associated with such births. Further we aimed to determine whether women at risk of accidental out-of-hospital delivery in our population could be identified antenatally. METHODS: A retrospective case-control study was performed. Women who delivered accidentally out-of-hospital in our catchment area between January 1995 and March 1999 were identified (cases) and compared with women who delivered in hospital following spontaneous labor (controls). Outcome measures included maternal demographic characteristics, obstetric features and neonatal outcome. RESULTS: In the study period, 117 women delivering 121 babies were identified who delivered accidentally out-of-hospital, (0.6% of all deliveries registered at the hospital). Women who delivered before arrival at hospital were more likely to be of greater parity, unbooked, late bookers and/or poor attenders for antenatal care. Gestation at delivery, duration of labor and birthweight were less in the out-of-hospital delivery group compared with the control group. The rate of perineal suturing was lower for cases than controls. Babies who were delivered accidentally out-of-hospital were more likely to require admission to the neonatal unit and had a higher perinatal mortality rate than controls (51.7 versus 8.6/1000 deliveries, respectively). CONCLUSIONS: Accidental out-of hospital deliveries account for less than 1% of deliveries in our population, but are associated with significant perinatal morbidity and mortality. Women should be educated regarding the importance of both antenatal care and a planned delivery. Since the majority of women who deliver accidentally out-of hospital are parous, there is an opportunity to do this in a previous confinement. PMID- 11942888 TI - Is it possible to recommend an "optimal" postoperative management after hysteroscopic metroplasty? A retrospective study with 52 infertile patients showing a septate uterus. AB - BACKGROUND: The aim of the retrospective study was to evaluate the "optimal" postoperative management after hysteroscopic metroplasty. METHODS. FIFTY-TWO: infertile patients with a septate uterus were included. Hysteroscopic metroplasty was performed using electrocautery (dissection needle) with postoperative cyclical hormone replacement therapy (HRT) + intrauterine device (IUD) insertion for 3 months (Group 1, n = 22), or HRT alone for 3 months (Group 2, n = 13), or without postoperative therapy (Group 3, n = 17). RESULTS: During a median follow up of 21+/-16.9 months 40.9% in Group 1, 53.8% in Group 2 and 41.2% in Group 3 (p > 0.05) resulted in ongoing pregnancies. The rates of delivery at term were 53.3%, 64.4% and 88.9%, respectively. CONCLUSIONS: A postoperative 3-months HRT + IUD insertion or a HRT alone after hysteroscopic metroplasty are not necessary. PMID- 11942889 TI - Two-step test: the combined use of fetal fibronectin and sonographic examination of the uterine cervix for prediction of preterm delivery in symptomatic patients. AB - PURPOSE: Our purpose was to assess the possibility of combined use of fetal fibronectin testing and sonographic examination of the uterine cervix for prediction of preterm deliveries. METHODS: We prospectively evaluated 82 patients with preterm labor. In each case fibronectin testing and sonographic examination of the cervix were performed. The primary outcome measure was preterm delivery within 28 days. RESULTS: Seventeen point one percent of the examined women delivered within 28 days. Among the analyzed sonographic parameters of the cervix only the functional canal length was significantly correlated with delivery (21.6+/-7.0 mm vs. 30.1+/-6.5 mm, p<0.001). Positive fetal fibronectin was found in 71.4% of patients delivered within 28 days and in 7.4% delivered after 28 days (p<0.001). In multiple logistic regression analysis only positive fibronectin testing (OR 11.25, p=0.005) and functional canal length < or =20 mm (OR 8.18, p=0.027) were independently associated with preterm delivery. We propose two-step testing: begin with the sonographic assessment of the cervix with functional canal length 31 mm as a negative one. The fibronectin testing should be performed only in patients with length 21-31 mm. Two step testing had overall sensitivity of 86%, specificity of 90%, positive predictive value of 63% and negative of 97% for predicting delivery within 28 days. CONCLUSIONS: The proposed combined use of the cervical sonography and fetal fibronectin testing is a practical diagnostic tool for predicting preterm delivery with higher sensitivity and negative predictive value than any of these methods alone. PMID- 11942890 TI - Reasons for pregnancy termination, contraceptive habits and contraceptive failure among Swedish women requesting an early pregnancy termination. AB - BACKGROUND: More than 30 000 legal abortions are performed every year in Sweden despite sexual education in schools, widespread youth-clinics and family planning services that are free of charge. The aim of this study was to investigate reasons for induced abortion, contraceptive habits and reasons for contraceptive failure among women presenting for induced abortion. METHODS: A questionnaire was administered to 591 Swedish-speaking women consecutively attending three different health care providers concerning an induced abortion during spring 2000. RESULTS: The response rate was 88% (n = 518). As many as 43%, among daily smokers 53%, had experienced one or more previous legal abortions. The majority of the women (97%) had discussed the decision about abortion with someone. The most cited reasons contributing to their decision were financial concerns, worries about the relationship and bad timing of the pregnancy. Though 85% had used contraception during the previous year, 36% of the women had not used any contraceptive method at the time of conception. The main reason given for not using contraception was the belief that they could not at that time become pregnant (35%). Ninety percent of the women planned to use contraception after the abortion. CONCLUSION: Women's decisions regarding induced abortion are multifactorial. One important reason was "poor economy". One out of three did not use any contraception, as they believed they could not become pregnant. Women presenting for induced abortion are a risk-group for further terminations. Counseling must include information about the fertile window, effective contraceptives and the emergency contraceptive pill. PMID- 11942891 TI - A nationwide analysis of complications associated with the tension-free vaginal tape (TVT) procedure. AB - BACKGROUND: To evaluate the therapy-associated morbidity of all patients who underwent a TVT operation in Finland by the end of the year 1999. METHODS: Questionnaires on the number of operations and on the number of different complications were sent to 38 hospitals where TVT operations had been independently performed after an obligatory TVT training period. The primary TVT training center and a hospital, which did not use the standard TVT equipment, were excluded. RESULTS: Among the 38 hospitals there were four university, 13 central and 21 local hospitals. The total number of operations was 1455. The incidence of bladder perforation was 38/1000, that of intra-operative blood loss over 200 ml 19/1000, of major vessel injury 0.7/1000, of nerve injury 0.7/1000, of vaginal hematoma 0.7/1000 and of urethral lesion 0.7/1000. The incidence of minor voiding difficulty was 76/1000, that of urinary tract infection 41/1000, of complete postoperative urinary retention 23/1000, of retropubic hematoma 19/1000, of wound infection 8/1000 and of vaginal defect healing 7/1000. No case of tape rejection or life threatening complication occurred and the incidence of complications requiring laparotomy was 3.4/1000. The ratio of number of complications to TVT operations performed did not vary significantly between different hospital types (p>0.05). CONCLUSION: The TVT procedure is a safe method for the treatment of stress urinary incontinence provided that appropriate training is offered. PMID- 11942892 TI - Is peritoneal closure necessary in obstetrical and gynaecological surgery? PMID- 11942893 TI - Closure of the peritoneum is not necessary in obstetric and gynecologic surgeries. PMID- 11942894 TI - Large paravaginal solitary fibrous tumor with secondary schistosoma hematobium infestation. PMID- 11942895 TI - Unruptured pregnancy in a non-communicating heterotopic right fallopian tube associated with left unicornuate uterus: evidence for transperitoneal sperm and oocyte migration. PMID- 11942896 TI - Relation of vitamin D receptor FokI start codon polymorphism to bone mineral density and occurrence of osteoporosis in postmenopausal women in Taiwan. AB - BACKGROUND: Osteoporosis is a common disorder with a strong genetic component. Our aim was to evaluate the correlation of the vitamin D receptor FokI start codon polymorphism to bone mineral density and the occurence of osteoporosis. METHODS: We determined the vitamin D receptor FokI start codon polymorphism using polymerase chain reaction-based restriction analysis in 163 postmenopausal women in Taiwan. The vitamin D receptor gene polymorphism was detected by the restriction enzyme FokI, where the F allele indicated the absence of the cuttable site and the f allele its presence. We then related the genotypes to bone mineral density and the occurence of osteoporosis in these women. RESULTS: The allelic frequencies for 163 postmenopausal women in Taiwan were 59.2% for F and 40.8% for f in FokI restriction fragment length polymorphisms. The prevalence of each genotype in the study population was: 42.3% FF, 33.7% Ff and 24% ff. The three genotypic groups differed significantly in bone mineral density at the lumbar spine (P = 0.029). Bone mineral density was highest in the Ff group and lowest in the ff group at the lumbar spine and the femoral neck. The FokI vitamin D receptor genotype showed a significant effect on the prevalence of osteoporosis in the subjects at the lumbar spine. That is, women with genotype ff had a 2.8 times greater risk for osteoporosis (P < 0.05), and those with genotype FF had a 0.8 times greater risk than women with genotype Ff. CONCLUSION: Our findings indicate that the vitamin D receptor FokI start codon polymorphism is associated with reduced bone mineral density and predisposes women to osteoporosis at the lumbar spine. PMID- 11942897 TI - Intra-hepatic cholestasis of pregnancy in hepatitis C virus infection. AB - BACKGROUND: Aims of this study were to investigate whether hepatitis C virus infection influences the incidence and natural history of intrahepatic cholestasis of pregnancy (ICP) and whether ICP has different characteristics in hepatitis C virus (HCV) positive women from ICP in HCV negative women. METHODS: A prospective study for the prevalence of the HCV infection and for the incidence of ICP was carried out in the 5840 patients admitted to the Prenatal Department of Padua University, Italy, between January 1996 and January 1999. Testing was done for HCV by the enzyme linked immunosorbent assay (ELISA 3), recombinant immuno blot assay (RIBA 3) and polymerase chain reaction (PCR). The diagnosis of ICP was made on clinical grounds based on the occurence of pruritus with onset during pregnancy, persisting up to the time of delivery and disappearing after delivery, supported by demonstrating an elevation of both serum ALT and total serum bile acids. The Student's t-test, one way anova and chi-square tests were used for statistical analysis. RESULTS: During the study period, 56 of 5840 patients developed ICP (0.96%). Of these, 12 were also HCV-RNA positive. The rate of ICP was observed more commonly in HCV-RNA positive women than in HCV-RNA negative women (20.33% or 12/59 versus 0.78% or 44/5767, P = 0.001 CONCLUSIONS: Occurrence of ICP during the third trimester should be an indication to investigate the HCV status of the patient. Although the diagnosis of ICP is not confirmed by specific tests, we confirmed a higher risk of HCV infection in this condition. Therefore, occurence of ICP during the third trimester should be an indication to investigate the HCV status of the patient. Broader studies are necessary to assess the impact of infection on the perinatal outcome of ICP. PMID- 11942898 TI - Newly delivered women's evaluation of personal health status and attitudes towards sickness absence and social benefits. AB - BACKGROUND: Unexpectedly high rates of sickness absence have been observed among pregnant women. No clear medical causes for illnesses reported as the basis for sick leave have yet been identified with certainty. An explanation proposed is the pregnant women's own attitudes towards their own states of well being during pregnancy. The aim of this study was to investigate the validity of this hypothesis. METHODS: All of the 384 women who were delivered at the University Hospital during a 2-month period were asked to answer a questionnaire anonymously. Information was sought concerning sickness absence and the use of parental benefits. In addition, questions were asked about working conditions and about each mother's own estimate of her level of 'well being'. The women's attitudes towards work absence due to illness and towards social benefit programs were registered. RESULTS: Forty-three per cent of the women stated that they had been on sick leave during pregnancy. The main reason for sick leave was reported back pain. Seventy-four per cent of the women who were on sick leave stated, nevertheless, that they had been in 'good' or 'excellent' health during pregnancy. Of the 149 women who did not take sick leave, 10 reported being in 'bad' or 'very bad' health during pregnancy. 4.3% of the women stated that they had considered themselves to be ill due to an obstetric condition. CONCLUSION: In addition to actual disease and severe discomfort, certain social conditions and attitudes as well, are likely to explain the increase of pregnant women on sick leave. PMID- 11942899 TI - The fetal origins hypothesis: placental insufficiency and inheritance versus maternal malnutrition in well-nourished populations. AB - The 'Fetal origins hypothesis' states that individuals born small because of malnutrition are predisposed to adult diseases. Fetal malnutrition has two main causes, poor maternal nutrition and placental insufficiency. A distinction between these causes is important because it is likely that maternal nutrition has been sufficient in the majority of populations in which the fetal origins hypothesis has been tested. Thus, placental insufficiency is a more reasonable cause of reduced fetal growth in adequately nourished populations. Placental insufficiency is mainly due to inadequate vascular adaptation at the uteroplacental interface ('poor placentation'). Among women with placental insufficiency syndromes such as pre-eclampsia and 'idiopathic' intrauterine growth retardation, there is an increased prevalence of risk factors for cardiovascular diseases. Maternal cardiovascular risk factors may therefore increase the risk of adult diseases in the offspring both through direct inheritance and by interfering with uteroplacental vascular adaptation. The latter may result in placental insufficiency and fetal growth retardation that by itself could cause adult disease (as the Fetal origins hypothesis states). Alternatively, the association between low birth weight for gestational and adult disease could be an epiphenomenon, leaving inheritance as the main explanation for the fetal origins hypothesis, in adequately nourished populations. PMID- 11942900 TI - Diabetic family history is an isolated risk factor for gestational diabetes after 30 years of age. AB - BACKGROUND: Family history of diabetes is regarded as a risk factor for gestational glucose intolerance. Most selective screening policy includes family history as an indication of oral glucose tolerance test. However, few studies had evaluated the actual incidence of glucose intolerance in this group of women. METHODS: Pregnant women with oral glucose tolerance test performed for family history of diabetes over two consecutive years were identified. Receiver operating curve was used to identify the age cut-off that give the highest sensitivity and specificity. The incidence of glucose intolerance was compared between women with age above and below this cut-off. RESULTS: One hundred and eighty-seven women were identified. The age cut-off selected was 30.5 years. In young women (age 60 mmHg, where k can be estimated from in vitro data and r=r0 at complete autoregulatory vasodilation. After introducing r=r(0)(1 + k - pk), Poiseuille's formula was integrated along the main segment, Deltax long, between arterial pressure P(1) and P(2) at the end of the main segment. At the lowest autoregulatory pressure P(1)=65 mmHg pre-glomerular blood flow is F=5Kr(0)(4)/Deltax. At P(1)=140 mmHg a pressure drop of only 17 mmHg to P2=123 mmHg is sufficient to fulfil the criterion for complete autoregulation: F=5Kr(0)(4)/Deltax. Thus, 80% of the total pre-glomerular vascular resistance is localized to the juxtaglomerular segment. Loop diuretics may abolish juxtaglomerular contractility. Calculated flow/pressure relationships after eliminating juxtaglomerular contractility are similar to those obtained after administering ethacrynic acid. If a constant tension hypothesis (r=60r(0)/p) rather than the transmural pressure hypothesis [r=r(0)(1 + k - pk)] applies, complete autoregulation is maintained to P(2)=89 mmHg, but the effect of loop diuretics is not mimicked. In conclusion, high juxtaglomerular contractility may be attributed to a myogenic mechanism only if extravascular pressure in the juxtaglomerular segment is subatmospheric. PMID- 11942923 TI - Analysis of the tubuloglomerular feedback mechanism in renal autoregulation. AB - Along the juxtaglomerular segment of the afferent arteriole the luminal pressure p approaches the glomerular capillary pressure of 55-60 mmHg. At such low luminal pressures the myogenic mechanism contracts only if extravascular pressure p(ex) is subatmospheric. According to Poiseuille's formula complete autoregulation requires that blood flow is F=5Kr(0)(4)/Deltax at arterial pressures exceeding 65 mmHg; r(0) is the radius of the relaxed segment at transmural pressure p - p(ex) < or =60 mmHg, where p(ex) is the extravascular pressure; Deltax is the length of the main preglomerular segment, 10 times longer than the juxtaglomerular segment. Consistent with in vitro studies a myogenic mechanism may reduce the relaxed juxtaglomerular radius r(jx)=0.7r(0) by 40% at a transmural pressure of 140 mmHg. Fifty and 60% reductions are also considered. Integration of Poiseuille's formula shows that complete autoregulation of preglomerular blood flow requires negative extravascular pressures p(ex)= -90 to -55 mmHg dependent on contractile force. Negative pressure of this magnitude is generated by effective hyperosmolality <5 mOsm across the membrane separating cleft from pole cushion. Negative pressure stays constant at arterial pressures exceeding 90-110 mmHg, implying constant tubuloglomerular feedback, but approaches atmospheric pressure at lower arterial pressure, suggesting maintenance of blood flow by reduction in the glomerular filtration rate; a rise in macula densa concentrations [NaCl](md) by 0.15 mM or [NaHCO(3)](md) by 2 mM raises extravascular pressure towards atmospheric levels by approximately 40 mmHg. A 40-mmHg rise in interstitial pressure exerts the same effect. Loop diuretics nullify osmotic force and dilate juxtaglomerular and main segments by raising juxtaglomerular extravascular pressure towards atmospheric levels. PMID- 11942924 TI - Respiratory mechanics in mice: strain and sex specific differences. AB - To assess the contribution of genetic background to respiratory mechanics, we developed a ventilator unit to measure lung function parameters in the mouse. We studied two commonly used inbred mice strains originating from Mus musculus domesticus (C57BL/6 and C3HeB/FeJ) and a third strain derived from Mus musculus molossinus [Japanese fancy mouse 1 (JF1)]. The ventilator allows for accurate performance of the different breathing manoeuvres required for measuring in- and expiratory reserve capacity, quasi-static and dynamic compliance, and airway resistance. In combination with a mass spectrometer for monitoring gas concentrations, single-breath manoeuvres were performed and He-expirograms obtained, from which dead space volume and slope of phase III were determined. From each strain and each sex, 10, 2-month old animals were studied immediately after being killed by an intraperitoneal overdose of xylazine and ketamine. C3HeB/FeJ and C57BL/6 exhibited comparable lung volumes. In male C3HeB/FeJ mice, e.g. vital capacity (VC) was 1072 +/- 79 microL, inspiratory reserve capacity 782 +/- 88 microL, and dead space volume at total lung inflation 216 +/- 18 microL. Lung volumes of JF1 were significantly lower (e.g. VC 611 +/- 53 microL, P < 0.01) even when normalized to body weight. In all three strains, specific lung volumes were significantly higher in females than in males, possibly explained by a higher oxygen demand during pregnancy and lactation, both of which fill most of their life times. Static compliance in C3HeB/FeJ was 64.3 +/- 5.4 microL cmH2O-1. It was smaller in C57BL/6 and JF1 mice, even when related to the lung volume. Analysis of the degree of genetic vs. non-genetic components of the phenotypic variation revealed that at least 80% of the total variation of lung volumes and static compliance in the mixed population is attributable to genetic differences between individuals. These differences will be verified in further studies by segregation and genetic linkage analysis. PMID- 11942925 TI - Intra- and peri-tendinous microdialysis determination of glucose and lactate in pigs. PMID- 11942927 TI - Randomized controlled trials in schizophrenia: a critical perspective on the literature. AB - OBJECTIVE: The randomized trial provides an opportunity to minimize the inclusion of biases in the evaluation of interventions in psychiatry. Difficulties arise, however, when applying their results to 'real world' clinical practice and decision-making. We, therefore, examined the real world applicability of schizophrenia trials. METHOD: A narrative overview of the content and quality of the randomized trials relevant to the care of those with schizophrenia is provided. RESULTS: Complex, explanatory, under-powered randomized drug trials dominate evaluative research in schizophrenia. CONCLUSION: Explanatory designs are a necessary but insufficient step in establishing the true worth of interventions in schizophrenia. Research from other spheres of mental health and wider health care suggest that pragmatic trials are feasible. This design allows large scale trials to be conducted which include patients which we would recognize from routine practice and which record outcomes which are of genuine interest to decision-makers. PMID- 11942928 TI - Screening for major depression in Asian-Americans: a comparison of the Beck and the Chinese Depression Inventory. AB - OBJECTIVE: This study compares the effectiveness of the Chinese version of the Beck Depression Inventory (CBDI) and the Chinese Depression Inventory (CDI) in screening for depression among Chinese Americans. METHOD: Five hundred and three Chinese-Americans in primary care were administered the CBDI and the CDI for depression screening. The results were compared with standard semistructured interview. RESULTS: With empirically determined cutoff scores of the CBDI (> or =13) and the CDI (>/=16), both instruments have good sensitivity (0.78) and excellent specificity (0.91 and 0.93, respectively). The correlation between the total scores of the two instruments was high (0.785, P < 0.01). The areas under the Receiver Operating Characteristic (ROC) curve of the CBDI and the CDI were 0.94 and 0.95, respectively, and were not significantly different. CONCLUSION: When administered by interviewers, the CBDI and the CDI have comparable effectiveness. Low participation among Chinese-Americans with self-report measures limits both scales as efficient depression screening instruments. PMID- 11942929 TI - Increased incidence of affective disorders, anxiety disorders, and non-natural mortality in women after breast cancer diagnosis: a nation-wide cohort study in Denmark. AB - OBJECTIVE: To investigate whether breast cancer patients have increased incidence of psychiatric admission with affective disorders, anxiety disorders, or non natural mortality compared with the general female population. METHOD: Register linkage between nation-wide registries: The Danish Psychiatric Central Register, The Danish Cancer Registry, and The Danish National Register of Causes of Death. A total of 61 709 women registered with primary invasive breast cancer between 1970 and 1993 were included and 356 023 person-years were accrued. RESULTS: The standardized incidence ratio of first-ever psychiatric admission with affective disorder was 1.49 (95% CI: 1.35-1.63) and with anxiety disorder 1.25 (95% CI: 1.06-1.46). The standardized non-natural mortality ratio during the first year after breast cancer diagnosis was 1.54 (95% CI: 1.27-1.87). All analyses were adjusted for age, calendar period, and place of residence. CONCLUSION: Breast cancer patients have significantly increased incidence of psychiatric admission with affective disorders, anxiety disorders, and non-natural mortality. PMID- 11942931 TI - Elderly suicide rates in Asian and English-speaking countries. AB - OBJECTIVE: Asian culture venerates elderly people. It was hypothesized that elderly suicides would be proportionately lower in Asian societies than in English-speaking countries (ESC). METHOD: Elderly (i.e. aged 75 years or more) to general population suicide ratios were compared for six Asian societies and six ESC, based upon the latest 5-year suicide rates. RESULTS: Males: The general population suicide rate was highest in rural China (227 per million) and third highest in Japan (217 per million), but other countries with high rates were all ESC (ranging from 224 to 198 per million). Asian countries had the six highest elderly suicide rates, 1327-1373 per million, whilst the highest ESC elderly rate was in the United States (507 per million). Asian societies had the widest range of elderly/general ratios, ranging from 6.62 to 2.6. Females: The overall suicide rates were higher in Asian countries (57-95 per million) than in ESC (40-56 per million). Asian elderly suicide rates differed (932 per million in rural China to 154 per million in Korea) but the highest ESC rate was in Australia (76 per million). The Asian countries elderly/general ratios ranged from 5.82 : 1 to 2.70 : 1, but the widest ESC ratio was 1.71 : 1, in the United Kingdom. CONCLUSION: There is a need for country-specific prevention measures in elderly people, and particularly for older women in Asian countries. PMID- 11942930 TI - Age, period and cohort effects on suicide rates in Australia, 1919-1999. AB - OBJECTIVE: To examine age, period and cohort effects on Australian suicide rates. METHOD: Male suicide rates for successive 5-year periods between 1919 and 1998, and for 1999 were displayed graphically to examine interactions between age, period and cohort effects. RESULTS: There has been a pronounced period effect on male suicide rates in all age groups over the last few decades, with lower rates in wartime and peak rates for most cohorts in the 1960s. Peak rates of all adult female 5-year age cohorts occurred in the 1960s or early 1970s. CONCLUSION: Most so-called cohort effects appear to be associated with environmental changes that may not be a function of the cohort itself. While much attention has focused on the rising suicide rates in young males in Australia, local media and health authorities have given little emphasis to the high rates found in elderly males. PMID- 11942932 TI - Do cenesthesias and body image aberration characterize a subgroup in schizophrenia? AB - OBJECTIVE: To identify and characterize a subgroup of schizophrenia patients with marked and dominating bodily sensations (cenesthesias and body image aberration). METHOD: We assessed cenesthesias and different aspects of body image aberration systematically along with common (general, positive and negative symptoms) and ego-psychopathology in 60 patients with acute paranoid schizophrenia. Cluster analysis was applied to identify subgroups. Psychopathology scores of the clusters were compared at admission and after 2 weeks of hospital treatment. RESULTS: One of the three clusters comprised of 14 patients (23.3%) with marked disturbances of body experience (underestimation of lower extremities, desomatization, boundary loss and diminution). The subgroup was further characterized by significantly higher ego-psychopathology scores at admission. Some of the differences held true over a 2-week period. CONCLUSION: The findings from the present study suggest that cenesthesias and body image aberration might represent an additional dimension of psychopathology that might be used for defining a nosological subtype of schizophrenia. PMID- 11942933 TI - Quality of life in patients with schizophrenia in five European countries: the EPSILON study. AB - OBJECTIVE: To compare subjective quality of life (QOL) and objective QOL indicators in patients with schizophrenia from five European sites: Amsterdam, Copenhagen, London, Santander and Verona. METHOD: A representative sample of 404 patients with schizophrenia, in contact with mental health services, was randomly selected and evaluated with the Lancashire Quality of Life Profile (EU). RESULTS: The level of satisfaction in certain domains, religion, family and social relations appears to be associated with local style of living and culture while work, finances, and safety were more independent from local variations. In addition to the severity of symptoms, frequency of contacts with family, friendship and age appear as predictors of QOL, all of them influenced by the characteristics of the surroundings. CONCLUSION: The centres participating in the study presented differences in subjective measures of QOL, objective indicators and also in service provision and styles of living. PMID- 11942934 TI - Quality of life in schizophrenia measured by the MOS SF-36 and the Lancashire Quality of Life Profile: a comparison. AB - OBJECTIVE: To compare two Quality of Life (QoL) instruments on reliability, feasibility and conceptual overlap in a group of schizophrenic out-patients. METHOD: The Lancashire Quality of Life Profile (LQoLP) and the MOS SF-36 were used to assess the QoL of 143 schizophrenic out-patients. RESULTS: Feasibility and reliability for both instruments were satisfying. Second order factor analysis on 10 LQoLP and eight MOS SF-36 scales resulted in three factors: one health related QoL factor and two general QoL factors; an internal and an external factor. CONCLUSION: QoL measures in schizophrenia studies are not exchangeable. Validity of a specific QoL instrument depends upon the purpose of the study. The LQoLP allows suggestions for specific improvements in mental health care for long-term psychiatric patients. The SF-36 is a good choice when comparison with other patient groups on health related QoL is relevant. PMID- 11942935 TI - Child related strain and maternal mental health: a longitudinal study. AB - OBJECTIVE: To explore how normal age-related behaviour patterns and medical status affect maternal mental health in children aged 1.5-4 years. METHOD: Data were collected via questionnaires from a population-based sample. Outcome variable was the Hopkins Symptom Checklist (HSCL-25). Multiple regression analyses were used to assess the extent to which changes in the predictors (sociodemographic variables, chronic strains, negative life events, maternal somatic health and social support) explained changes in maternal distress. RESULTS: Changes in strain related to children and child care-taking predicted changes in maternal mental distress stronger than the other explanatory variables. Only effects of changes in child care-taking were significant for both time periods. CONCLUSION: Changes in maternal symptoms of anxiety and depression appear to be influenced by changes associated with children's behaviour, their medical status and child care-taking arrangements. PMID- 11942936 TI - Recollections of parent-child relationships in patients with obsessive-compulsive disorder and panic disorder with agoraphobia. AB - OBJECTIVE: In previous studies, patients with different psychiatric conditions, as compared with matched controls, have reported that their parents were more protective and less caring towards them when they were children. However, studies investigating associations between parental behaviours and anxiety disorders have yielded inconsistent results. The aim of this study was to compare recalled parental behaviours in out-patients with obsessive-compulsive disorder (OCD), in out-patients with panic disorder with agoraphobia (PDA), and in non-anxious controls. METHOD: The sample included 43 out-patients with OCD, 38 with PDA, and 120 controls. Participants completed the Parental Bonding Instrument and the Egna Minnen Betraffande Uppfostran or Own Memories of Parental Rearing Experiences in Childhood. RESULTS: No differences were found between the two anxious groups. However, compared with the control group, anxious patients recalled their parents as more protective. CONCLUSION: Our findings suggest that child rearing practices such as overprotection may be a risk factor in the development of anxiety disorders. PMID- 11942937 TI - Age-at-onset and schizophrenia: reversed gender effect. AB - OBJECTIVE: This study seeks an explanation for reversed gender effect on age-at onset (AAO) in schizophrenia. The hypothesis is older AAO in males would be detected in a sample where higher infant mortality (IMR) prevailed. METHOD: Case records of International Classification of Diseases-10 (ICD-10) schizophrenia patients from two states (n=70 each) with an IMR of 13 and 67 per thousand were reviewed and AAO was obtained by using the recorded age and duration of illness. RESULTS: In the sample from the state with lower IMR, AAO did not differ between the two sexes. However, men had older AAO than women in the state with fivefold higher IMR. CONCLUSION: Gender differences in AAO may be a function of perinatal complications. In places where infants with perinatal complications are less likely to survive, hence high IMR, a small group of potentially youngest AAO schizophrenic males may be eliminated thus changing the gender effect on AAO. PMID- 11942938 TI - Internationalization of psychiatric research - the prospective for the European Association of Psychiatrists. PMID- 11942939 TI - The application of transcranial magnetic stimulation in psychiatry and neurosciences research. AB - OBJECTIVE: Over recent years transcranial magnetic stimulation (TMS) has become widely applied in the study of neuropsychiatric disorders. The aim of this article is to review the application of TMS as an investigative tool and as a potential therapeutic modality in psychiatric disorders. METHOD: A comprehensive literature review. RESULTS: When applied as an investigative tool, TMS provides innovative ways to directly study the excitability of the cortex, cortical regional connectivity, the plasticity of brain responses and cognitive functioning in illness and disease states. A number of studies suggest the potential of treatment with TMS in disease states, especially in patients with depression, although difficulties exist with the interpretation of the published literature. CONCLUSION: TMS has a considerable role in neuropsychiatric research. It appears to have considerable potential as a therapeutic tool in depression, and perhaps a role in several other disorders, although widespread application requires larger trials and establishment of sustained response. PMID- 11942940 TI - Self-rated schizotypal cognitions, psychotic symptoms and the onset of schizophrenia in young people at high risk of schizophrenia. AB - OBJECTIVE: To examine the associations between the Rust Inventory of Schizotypal Cognitions (RISC) and psychotic symptoms measured by the Present State Examination (PSE) and to assess the predictive validity of the RISC for later onset of schizophrenia. METHOD: A total of 154 subjects at high risk for schizophrenia but who were currently well, 35 well controls and 28 first episode schizophrenic patients were assessed on specified PSE symptoms and completed the RISC. A subsample of the high risk group was subsequently monitored for onset of schizophrenia over 39 months. RESULTS: RISC scores at entry were significantly higher in subjects with psychotic symptoms on PSE. There were indications that high scorers on the RISC were likely to develop schizophrenia. However, less than half of those falling ill exhibited high RISC scores on entry. CONCLUSION: The RISC scale is highly associated with presence of psychotic symptoms on PSE and high scorers may be at increased risk of schizophrenia onset. PMID- 11942942 TI - Religious coping and psychological wellbeing in carers of relatives with schizophrenia. AB - OBJECTIVE: To examine the use of religious coping and its relation to psychological wellbeing in carers of relatives with schizophrenia. METHOD: Sixty carers of patients with an ICD-10 diagnosis of schizophrenia, were assessed on strength of religious belief, perceived burden, religious and other coping strategies and psychological wellbeing. RESULTS: Coping strategies of denial and problem solving, strength of religious belief and perceived burden were significant predictors of wellbeing. CONCLUSION: Strength of religious belief plays an important role in helping family members to cope with the stress of caring for a mentally ill relative. In addition to psychoeducation and problem solving coping, the role of religious coping in enhancing wellbeing of carers needs to be considered in family intervention programmes. PMID- 11942941 TI - Comparisons between schizophrenia patients recruited from Australian general practices and public mental health services. AB - OBJECTIVE: To examine differences between samples of schizophrenia patients recruited from general practice and public mental health services. METHOD: Demographic, psychosocial, disability and 12-month service utilization data are reported from a multicentered survey of psychotic disorders and an associated study of schizophrenia in general practice. Patients with schizophrenia from three recruitment sources (in-patient, community services, general practice) were compared. RESULTS: General practice patients had fewer symptoms, better functioning, lower service use, but comparable substance abuse, to patients from mental health services. They were generally similar to community mental health patients, with the exception of family support, premorbid work adjustment, negative symptoms and disability. Service contact models are also reported which demonstrate that general practitioners deal with schizophrenia patients across the range of illness severity and acuity. CONCLUSION: Recruitment source impacts in schizophrenia research need to be more carefully considered during sample selection and better accounted for in the interpretation of results. PMID- 11942943 TI - Development of specific social skills training programmes for schizophrenia patients: results of a multicentre study. AB - OBJECTIVE: The efficacy of three newly developed cognitive social skills training programmes for residential, vocational and recreational functioning (experimental groups) were compared with a traditional social skills training programme (control group) referring to cognitive and social abilities, psychopathology and generalisation effects. METHOD: One hundred and five patients with a diagnosis of schizophrenia or schizoaffective disorder according to ICD-10 criteria were selected and assigned to the different treatment groups, using a matching procedure. The treatment phase lasted 6 months. A follow-up assessment was carried out after 1 year. RESULTS: Higher global therapy effects were obtained on almost all dependent variables in the experimental groups. Analyses of variance and covariance indicated higher symptom reduction for the experimental groups, but significantly greater improvements in some cognitive variables for the control group. Correlation analysis suggested associations between improvement of social behaviour with symptom reduction and improvements of cognitive skills. CONCLUSION: In view of these favourable effects, the developed cognitive social skills training programmes might facilitate the abilities of schizophrenia patients for their integration in the community. PMID- 11942944 TI - Social desirability and eating disorders. A community study of an Italian school aged sample. AB - OBJECTIVE: To explore the links between social desirability and eating disorders in a sample of adolescents in a north-east area of Italy. METHOD: A mixed male female sample of 1000 school-aged adolescents, corresponding to 10% of the young population aged 15-19 years living in the district, were investigated with self reported questionnaires, including the Eating Attitudes Test (EAT), the Bulimic Investigatory Test of Edinburgh (BITE), the Body Attitudes Questionnaire (BAT), and an Italian version of the Marlowe-Crowne Social Desirability Scale (MC-SDS). RESULTS: Females scored higher than males at all eating disorder inventories. In both genders there was a negative relationship (in all cases P < 0.01) between scores at the eating disorder inventories and those at the MC-SDS. When analysing eating disorder "caseness", as measured by cut-off, "cases" reported significantly lower scores than "non-cases" at the MC-SDS in both genders. CONCLUSION: Personality traits measured by the MC-SDS, such as defensiveness, self-esteem, and dependence from approval, might contribute to the development of abnormal eating patterns at risk of eating disorders. PMID- 11942945 TI - Seasonal affective disorder in an Arctic community. AB - OBJECTIVE: To determine the rate of seasonal-pattern depression in an Inuit community above 70 degrees N. METHOD: One-hundred and eleven people from randomly selected households were surveyed for depression and anxiety and the effect of the seasons on their mood. Eighty-eight people provided replies on the influence of the seasons. RESULTS: One in five (22.6%) of the community sample was found to be depressed. Of these, seven (6.3%) had seasonal affective disorder (SAD), with fall onset occurring in six of these (5.4%). Subsyndromal SAD (SSAD) occurred in 11.7%, while any effect of the seasons (seasonality) occurred in 39.6%. Persons with SSAD and seasonality were significantly older than those unaffected by the seasonal effect. No other significant correlation of SAD, SSAD, or seasonality occurred with gender, age, and language preference. CONCLUSION: Seasonal mood changes in this Inuit group living in the Canadian Arctic are elevated above the rates found in other studies using similar survey methods. PMID- 11942946 TI - Public attitude to compulsory admission of mentally ill people. AB - OBJECTIVE: Assessing the public attitude to compulsory admission of mentally ill in Switzerland and analyzing the influence of demographic, psychological, and sociological factors. METHOD: We conducted a representative telephone survey with 1737 interviewees. In a logistic regression analysis compulsory admission was used as the dependent variable. RESULTS: More than 70% of the respondents display a positive attitude to compulsory admission. Education, negative stereotypes, and living in the French speaking part of Switzerland are predictors for accepting compulsory admission whereas older age, extreme political opinion, and rigid personality traits show an inverse effect. Negative emotions, anomie, social distance, and contact to mentally ill have no significant influence. CONCLUSION: The mostly positive attitude to compulsory admission suggests that the public trust in psychiatry to deal with the assigned responsibility. PMID- 11942947 TI - Prediction of aggression on a locked psychiatric admissions ward. AB - OBJECTIVE: The present study evaluates the accuracy of clinical and archival predictors of patients' aggressive behaviour on a locked admissions ward. METHOD: Over a 9-month period, staff members estimated the likelihood that patients would become aggressive during their stay in the ward. These unaided clinical assessments were obtained with Visual Analogue Scales (VASs) administered before the end of the first full day of admission. Archival predictions were based on demographic variables (e.g. gender, number of previous admissions, diagnosis) derived from patients' admission forms. Aggressive behaviour was recorded with the Staff Observation Aggression Scale-Revised (SOAS-R). RESULTS: Clinical predictions of aggression were found to be moderately accurate. On the basis of clinical estimates, 75% of the patients were correctly classified as becoming aggressive or not. CONCLUSION: Although a body of evidence indicates that unaided clinical prediction of violent recidivism after hospital discharge does not perform well, it may be quite accurate in estimating short-term aggression risks during acute psychiatric admission. PMID- 11942948 TI - Dopamine transporter gene (DAT1) VNTR polymorphism in major psychiatric disorders: family-based association study in the Bulgarian population. AB - OBJECTIVE: A 40-bp variable number tandem repeat in the 3'-UTR of dopamine transporter gene (DAT1) has been examined for association with major psychiatric disorders in several case-control studies. No significant results have been found. We used a new collection of parent-offspring trios to test for association with schizophrenia (SZ), bipolar 1 disorder (BPI) and schizoaffective (SA) disorder. METHOD: We genotyped trios from Bulgarian origin where the proband had SZ (178 trios), BPI (77 trios) and SA (29 trios). Alleles ranging from 5 to 11 repeats were observed. The results were analysed with the extended TDT (ETDT). RESULTS: No preferential transmission of alleles was observed for any diagnostic group. The presence of allele DAT*10 was associated with the severity and frequency of auditory hallucinations, however, this result is not significant if corrected for multiple testing. CONCLUSION: Our results are in agreement with previous reports of a lack of association between this polymorphism and major psychiatric disorders. PMID- 11942949 TI - Wednesday, 17 April 2002.- Pre-congress symposium: The best of Nordic psychiatric epidemiology. PMID- 11942950 TI - Thursday, 18 April 2002. - Plenary lecture. PMID- 11942951 TI - Friday, 19 April 2002. - Plenary lecture. PMID- 11942952 TI - Saturday, 20 April 2002.- AEP epidemiology award lecture. PMID- 11942953 TI - Radiation hazards in urological practice. PMID- 11942954 TI - Optimizing medical radiation exposures for uroradiological procedures, with special emphasis on paediatric imaging. PMID- 11942955 TI - The need for routine adrenalectomy during surgical treatment for renal cell cancer: the Hannover experience. AB - OBJECTIVES: To further clarify the need for routine adrenalectomy during the surgical treatment of renal cell cancer, as in the absence of clinically overt metastatic disease, tumorous lesions within the adrenal gland are found in only 2 10% of patients, with most being over-treated by adrenalectomy. PATIENTS AND METHODS: The medical records of 819 patients undergoing adrenalectomy combined with nephrectomy, irrespective of the local extension of the primary tumour or the clinical stage at first diagnosis, were reviewed to determine the reliability of currently available imaging methods in predicting adrenal gland metastases. Several patient and tumour characteristics were correlated with the presence of intra-adrenal metastases, and their possible independent prognostic value was determined by a multivariate logistic regression model. RESULTS: There was metastatic spread into the adrenal gland in 27 of 819 (3.3%) patients. In only three of eight patients in whom the adrenal was identified as the only metastatic site were preoperative abdominal computed tomography scans interpreted as false negative. On multivariate statistical analysis only the presence of distant metastases, vascular invasion within the primary tumour and multifocal growth of renal cell cancer within the tumour-bearing kidney were identified as independent predictors of the presence of intra-adrenal metastases. CONCLUSIONS: None of the patient or tumour characteristics evaluated reliably predicted the likelihood of adrenal metastases in patients with no evidence of disseminated metastatic spread. However, previously published data indicate that the frequency of metachronous metastases within the contralateral kidney (1.8-3.8%) is significantly higher than the risk of a preoperatively undetected isolated intra adrenal metastatic lesion when currently available imaging modalities are applied. Therefore, routine adrenalectomy should not be recommended if the preoperative radiological examinations are normal. PMID- 11942956 TI - Cytoreductive nephrectomy: is it a realistic option in patients with renal cancer? AB - OBJECTIVE: To evaluate the role of cytoreductive nephrectomy (CRN) in improving survival in patients with renal cell cancer. PATIENTS AND METHODS: The case-notes of 268 consecutive patients who presented to our specialized renal cancer clinic between 1998 and 2001 were reviewed. All patients with metastatic disease were assessed for CRN. If their primary tumour was considered operable, they were assessed further using the European Cooperative Oncology Group performance score; only patients with a performance score of 0 or 1 were considered for surgery. RESULTS: In all, 168 patients underwent nephrectomy with curative intent for M0 disease and 11 were treated conservatively. Ninety-four patients with M+ disease (mean age 65 years, range 38-80) were considered for CRN. Thirty-eight patients had an inoperable primary. Of the remaining 56 patients, 20 had a performance status of 0 or 1 and were offered CRN. CONCLUSION: Metastatic disease at presentation occurred in 34% of all patients referred; 40% patients had an inoperable primary and 38% had a performance score of > or =2. With an active policy of considering all patients for CRN, only 7% of those with renal cancer were suitable for this procedure. CRN is unlikely to have a significant effect on overall survival within a population of patients with renal cancer. PMID- 11942957 TI - The clinical relevance of sex hormone levels and sexual activity in the ageing male. AB - OBJECTIVES: To assess the changes in sex hormone levels with age and the relationship of sexual functioning to testosterone levels, evaluating serum testosterone levels and erectile function in men with lower urinary tract symptoms (LUTS). PATIENTS AND METHODS: The study included 213 men with LUTS (age range 31-78 years) who had no confirmed erectile dysfunction. Their serum total and free testosterone, and sex-hormone binding globulin (SHBG) levels were measured, and they completed the International Index of Erectile Function (IIEF) questionnaire. RESULTS: The total and free testosterone levels decreased and SHBG increased with age, but only the change in free testosterone and SHBG were statistically significant. The correlation with age was closer for free testosterone (r = - 0.356, P < 0.001) than for SHBG (r = 0.177, P = 0.010). Regression analysis of the five domain scores of the IIEF and three hormonal levels, after correcting for age, showed that free testosterone level was significantly correlated with erectile function (r = 0.2136, P = 0.005) and orgasmic function (r = 0.179, P = 0.020), but SHBG levels were significantly correlated only with orgasmic function (r = - 0.154, P = 0.046). Total testosterone levels showed no significant correlation with any of the five domains of the IIEF. CONCLUSIONS: Of the sex hormone levels, the change in free testosterone correlated most closely with ageing and had the closest correlation with sexual activity. Contrary to previous reports, free testosterone and SHBG levels were significantly correlated with orgasmic function and/or erectile function rather than sexual desire. A complete study of sex hormone levels is needed to evaluate patients with erectile dysfunction. PMID- 11942958 TI - A demographic profile of patients undergoing transurethral resection of the prostate for benign prostate hyperplasia and presenting in acute urinary retention. AB - OBJECTIVE: To evaluate, in a prospective study, the demographic profile of patients with benign prostate enlargement who presented in acute urinary retention (AUR). PATIENTS AND METHODS: The study comprised all patients admitted for transurethral resection of the prostate and categorised into two groups, i.e. those presenting in AUR or electively. The factors evaluated included the length of hospitalization, the patients' occupation, their duration of symptoms and reasons for not seeking treatment. RESULTS: There was no significant difference in the mean age and occupational status of the two groups but those in AUR had more complications and a longer hospital stay after surgery; 60% of these men had had their urinary symptoms for > 1 year. When asked why they did not seek treatment earlier, 35% reported fear of surgery, while 41% thought that their symptoms were a normal part of ageing. CONCLUSION: There is a need to raise the level of public awareness of benign prostatic enlargement because those who present with AUR incur excess morbidity and longer hospitalization that could otherwise be avoided through earlier treatment and elective surgery. PMID- 11942959 TI - The provision of transurethral prostatectomy on a day-case basis using bipolar plasma kinetic technology. AB - OBJECTIVE: To evaluate, in a prospective study, the potential for providing elective transurethral prostatectomy on a day-case basis, using new bipolar electrosurgical technology. PATIENTS AND METHODS: Forty men with confirmed bladder outlet obstruction caused by benign prostatic enlargement underwent plasma kinetic vaporization of the prostate (PKVP) in the Day Surgery Unit; they were scheduled to be discharged on the day of surgery with an indwelling catheter, returning for a trial removal of the catheter at 48 h, and followed up at 2 and 4 months after surgery. RESULTS: Thirty-four (85%) of the 40 patients were able to return home on the day of surgery. All patients, including the six who failed to return home as a day-case, successfully voided at 48 h; 78% felt the procedure was acceptable on a day-case basis. At 4 months the mean improvement in the International Prostate Symptom Score and Quality of Life score were 64% and 84%, respectively. The mean improvement in flow rate was 200-250%. CONCLUSION: PKVP effectively relieves prostatic obstruction, at least in the short-term (a longer term follow-up is required). The procedure can be delivered safely on a day-case basis and was very acceptable to the patients. PMID- 11942960 TI - Prognostic value of the Gleason score in prostate cancer. AB - OBJECTIVE: To investigate the prognostic value of the Gleason score in prostate cancer. PATIENTS AND METHODS: A consecutive series of 305 men with prostate cancer diagnosed at transurethral resection (1975-1990) and with no curative treatment was analysed. There was no assessment of prostate-specific antigen level during this period. The mean (range) age at diagnosis was 73.7 (52-95) years and the mean follow-up was 6.4 (0-22) years. The influence of Gleason score and the percentage of the specimen area with tumour (% cancer) on disease specific survival were assessed using Kaplan-Meier analyses. RESULTS: Of 305 cancers, 22% had a Gleason score of 4-5, 29% of 6, 18% of 7 and 32% of 8-10. At the follow-up, 89% of the men had died, of whom 42% had died from prostate cancer. The disease-specific 10-year survival was 56%. The disease-specific mean survival (DSMS) for Gleason score 4-5, 6, 7 and 8-10 was 20, 16, 10 and 5 years, respectively (P < 0.001). The DSMS did not differ significantly between Gleason 4 and 5 or between 8-10. There was a trend towards shorter survival for Gleason 4 + 3=7 (DSMS 9 years) than GS 3 + 4=7 (DSMS 13 years; P = 0.16). Gleason score and % cancer were independent predictors of DSMS (P < 0.001). CONCLUSION: The long-term prognosis of prostate cancer on deferred treatment is predicted well by the Gleason score. Four prognostic categories of prostate cancer are suggested, i.e. Gleason score 4-5, 6, 7 and 8-10. PMID- 11942961 TI - Does site-specific labelling and individual processing of sextant biopsies improve the accuracy of prostate biopsy in predicting pathological stage in patients with T1c prostate cancer? AB - OBJECTIVE: To evaluate whether individual labelling and processing of the sextant of origin improves the accuracy of prostate biopsy in predicting the final pathological stage after radical prostatectomy in patients with T1c prostate cancer. PATIENTS AND METHODS: The charts of 386 patients treated for prostate cancer by radical prostatectomy between January 1996 and June 1999 were reviewed. In all, 124 patients fulfilled the following inclusion criteria: no abnormality on digital rectal examination (DRE) or transrectal ultrasonography, a prostate specific antigen (PSA) level before biopsy of < or = 20 ng/mL, and prostate cancer diagnosed after one set of random sextant biopsies, with the cores being submitted in six separate containers individually labelled for the sextant of origin. RESULTS: Within this series of patients with a low tumour burden, the preoperative PSA, biopsy Gleason score and unilateral vs bilateral involvement were not significant predictors of disease extension. The percentage of positive cores and the number and topography of positive sextants were both statistically significant predictors of organ-confined disease. Although these two variables appeared to be statistically equivalent on a first analysis in the overall series, a subgroup of patients was identified who benefited from the complete topographical information, i.e. those 52 (42%) patients with a Gleason score of < 7, 25-75% positive biopsies and < or =3 positive sextants. CONCLUSION: These results support the individual labelling of biopsy cores in selected patients with a normal DRE and a moderately elevated PSA, as it helps to better predict the final pathological stage. This substantial benefit outweighs the additional effort by the pathologist. PMID- 11942962 TI - The role of (18)fluoro-2-deoxyglucose positron emission tomography in initial staging and re-staging after chemotherapy for testicular germ cell tumours. AB - OBJECTIVE: To investigate the role of 18fluoro-2-deoxyglucose positron emission tomography (FDG-PET) in the initial staging of clinical stage I and II nonseminomatous germ cell tumours (NSGCTs) and in re-staging (non)seminomatous GCTs after chemotherapy. PATIENTS AND METHODS: FDG-PET studies were undertaken in 50 patients. FDG uptake was interpreted visually and when possible the standardized uptake value was determined. A FDG-PET scan was taken in five patients with clinical stage I and in seven with stage II NSGCT. The scans were validated by histology. Stage I patients underwent a retroperitoneal lymph node dissection because of vascular invasion in the primary tumour. Thirty-eight scans were taken after completing chemotherapy (28 NSGCTs and 10 seminomatous GCTs), and validated by histology or clinical follow-up. RESULTS: In stage I NSGCT, FDG PET staging was equivalent to computed tomography (CT) staging. One small lesion, consisting of mature teratoma, was missed by both FDG-PET and CT. In stage II NSGCT, FDG-PET missed two lesions (mature teratoma and retroperitoneal mass with a small component of embryonal cell carcinoma) whereas CT correctly classified all. In 20 of 28 patients with NSGCT, histology was obtained after chemotherapy. In one of three patients with viable tumorous residual mass the FDG-PET scan was clearly positive; in four of 12 with mature teratoma and inflammation components retroperitoneally, the FDG-PET was also positive. In contrast, eight patients with solitary mature teratoma had a negative PET result. In four of five patients with necrosis after chemotherapy the PET result was correctly negative. All eight patients on surveillance had a negative PET scan and were free of disease at median (range) of 14 (8-18) months. Interestingly, of the 12 patients with a correct negative PET result, 11 had no mature teratoma in their primary tumour. Nine of 10 patients with SGCT were correctly staged. Two FDG-PET studies showed increased uptake; in one, a viable seminomatous mass was found and in the other there was inflammation in the residual mass. In all other patients the FDG-PET scan correctly predicted absence of viability in the residual mass. CONCLUSIONS: In primary staging, FDG-PET has no benefit over CT. In re-staging, a negative FDG PET result predicts fibrotic residual mass in seminomatous GCT. Moreover, it could be useful to predict fibrotic residual mass in NSGCT in those patients with no teratoma component in their primary tumour. PMID- 11942963 TI - The role of imipramine therapy in bladder exstrophy after bladder neck reconstruction. AB - OBJECTIVE: To assess the role of imipramine therapy after bladder neck reconstruction in patients with bladder exstrophy. PATIENTS AND METHODS: Seventeen children with bladder exstrophy who achieved partial continence after bladder neck reconstruction received imipramine (1.5-2 mg/kg body weight) for a mean duration of 9.5 months. Assessment before and after therapy included an objective classification of the continence status and artificial slow-fill cystometry. RESULTS: Eleven of the 17 patients had a good clinical response to imipramine, with an increase in the continent period to > 2 h, and in nocturnal continence. The urodynamic findings showed a significant improvement in the capacity, end-fill pressure, uninhibited contractions and '20 below' capacity. Only minor side-effects of the drug were reported. CONCLUSION: Imipramine has a role in patients who achieve partial continence after reconstruction, and who have a moderately small capacity bladder with poor compliance and uninhibited contractions. PMID- 11942965 TI - Functioning adrenocortical neoplasms in children. AB - OBJECTIVE: To describe the clinical features, treatment and outcome of functioning adrenocortical tumours in children, and to assess the value of tumour size as a marker for malignant potential. PATIENTS AND METHODS: Twenty-one children (mean age at presentation 4.9 years, range 0.6-11; 12 girls and nine boys) who presented with a functioning adrenal mass over a 29-year period (1972 2000) were assessed. Clinicopathological features and outcome information were obtained from a structured review of the case-notes. Before 1980 the diagnosis and staging was mainly obtained by intravenous pyelography and plain X-ray. Since 1980, ultrasonography, computed tomography and more recently magnetic resonance imaging were the methods of choice. RESULTS: The mean (range) duration of symptoms before diagnosis was 31 (2-108) months. The symptoms and signs at presentation comprised virilization (18 children), Cushingoid features (three), precocious puberty (three) and hypertension (four). Three children had malignant neoplasms, one presented with metastasis and of the other two, one died from the disease within 6 months, despite adjuvant chemotherapy and radiotherapy. The third girl is disease-free 11 years after complete excision of the neoplasm. Eighteen children had a benign neoplasm and all are alive and free of recurrence. In most of the children surgical extirpation was relatively simple, blood loss minimal, recovery uneventful and the hospital stay short (mean 5 days). The virilizing effects of all the neoplasms gradually resolved after surgery within a median (range) of 27 (6-108) months. CONCLUSIONS: Functioning adrenocortical neoplasms in children are rare, with a peak incidence in the first decade, are predominantly unilateral and more common in girls. Most are benign, but both benign and malignant lesions can present with virilizing and Cushingoid features. The malignant neoplasms have an extremely poor prognosis, especially if excision is incomplete, despite adjuvant chemotherapy and radiotherapy. Although there is an association between tumour size and malignancy, this cannot be used as a reliable individual discriminator. PMID- 11942966 TI - Innervation of congenitally hydronephrotic and normal porcine upper urinary tract. AB - OBJECTIVE: To investigate the intrinsic innervation of the upper urinary tract in congenitally hydronephrotic and normal Goettingen minipigs, using the whole-mount preparation technique. MATERIALS AND METHODS: Whole-mount preparations of hydronephrotic (two with bilateral ectopic ureters, one with left distal ureteric stenosis) and normal (three) porcine upper urinary tracts were examined by immunohistochemistry with tyrosine hydroxylase (TH) and neurofilament and by histochemical staining with NADPH-diaphorase and acetylcholinesterase. Staining results were evaluated using normal bright-field and confocal laser scanning microscopy. RESULTS: Neurofilament-, TH-immunoreactive and acetylcholinesterase positive nerve fibres and neuronal networks were identified in the adventitial, muscle and subepithelial layers of the whole upper urinary tract. An NADPH diaphorase-positive network was expressed in the subepithelial layer and less densely in the muscle layer. The general distribution of the identified neuronal networks was similar in hydronephrotic and normal upper urinary tracts, but the density of these neuronal networks was less in the former. The most striking observation was the absence or marked reduction of neuronal networks in the stenotic part of the ureter in the pig with left distal ureteric stenosis. CONCLUSION: Whole-mount preparations provide a method for assessing the three dimensional topography of neuronal networks in the different layers of the porcine upper urinary tract. Although the macroscopic differences between the hydronephrotic and normal porcine upper urinary tracts were striking, changes in the innervation pattern were less obvious, except in distal ureteric stenosis. PMID- 11942967 TI - Renal calcium phosphate and oxalate deposition in prolonged vitamin B6 deficiency: studies on a rat model of urolithiasis. AB - OBJECTIVE: To determine the effect on kidney oxalate-salt deposition of a prolonged diet which induced vitamin B6 deficiency in adult rats, as there are reports of the pathogenic involvement of vitamin B6 deficiency in the formation of renal calcium oxalate calculi. MATERIALS AND METHODS: The study comprised 24 6 month-old male albino Wistar rats; 12 were fed with a purified vitamin B6 deficient diet and the others provided with the same diet but supplemented with 6 mg/kg of vitamin B6. After 12 weeks, all rats were killed, and their kidneys fixed in formalin and routinely processed to paraffin for morphological examination; some fragments were fixed in glutaraldehyde and prepared for ultrastructural examination. From each rat consecutive sections of both kidneys were cut and stained with haematoxylin and eosin, periodic-acid Schiff, Sirius red and the Von Kossa method for calcium. Sections were examined in polarized light and by electron microscopy. RESULTS: The histopathological and ultrastructural features of the kidney of vitamin B6-deficient rats were those of tubular-interstitial nephritis, characterized by tubular atrophy, interstitial fibrosis and chronic inflammatory infiltration. Oxalate and phosphate crystals were present in the papillary and parenchymal connective tissue. Ultrastructural features confirmed severe tubular epithelial lesions and the presence of an interstitial and intraepithelial inflammatory infiltrate; there was mild interstitial fibrosis. None of these features were apparent in the kidney of control rats. CONCLUSIONS: Histopathological and ultrastructural data indicate that a prolonged vitamin B6-deficient diet may contribute to the formation and deposition of calcium phosphate and oxalate crystals, which lead to severe damage of the renal parenchyma. This phenomenon may occur not only in growing rats, which have more active protein metabolism and consequently higher vitamin B6 requirements, but also in adult rats. PMID- 11942968 TI - Cyclic AMP and cyclic GMP activate protein kinase G in cavernosal smooth muscle cells: old age is a negative factor. AB - OBJECTIVE: To investigate protein kinase G-I (PKG-I) expression and activation in cavernosal smooth muscle cells (CSMC) of young and old rats. MATERIALS AND METHODS: PKG-I expression in rat penis was examined by immunohistochemical staining, reverse transcription-polymerase chain reaction, and Western blot analysis. CSMC isolated from young (16-week-old) and old (28-month-old) rats were grown as monolayer cell cultures and treated with different dosages of cAMP or cGMP for different periods. Their proteins were then analysed for the expression of vasodilator-stimulated phosphoprotein (VASP), phosphorylated VASP (at serine 239), PKG-I, and protein kinase A (PKA). RESULTS: PKG-I expression was detected in the vascular and CSMC of the rat penis. There was little or no difference in the level of PKG-I expression between young and old rats. Treatment of CSMC with different dosages of cAMP or cGMP did not change the expression levels of VASP throughout the entire test period (up to 24 h). In contrast, the level of VASP phosphorylation at S239, i.e. the level of PKG-I activation, depended on the dosages of cAMP and cGMP and on the duration of treatment. Prolonged treatment (24 h) with either cAMP or cGMP resulted in down-regulation of both PKG-I and PKA. While cAMP and cGMP produced very similar results in nearly every aspect, there was a difference in one test, in which cGMP produced much less activated PKG-I than cAMP in the CSMC of 28-month-old-rats. CONCLUSIONS: For the first time we provide evidence for PKG-I activation in CSMC. Both cAMP and cGMP were capable of activating PKG-I in CSMC. Age seemed to compromise the ability of PKG-I in response to cGMP. PMID- 11942969 TI - Possible autocrine loop of the epidermal growth factor system in patients with benign prostatic hyperplasia treated with finasteride: a placebo-controlled randomized study. AB - OBJECTIVE: To analyse the expression of the epidermal growth factor (EGF) system in prostate tissue and secretions obtained from patients with benign prostatic hyperplasia (BPH) treated with or without finasteride (which primarily targets the androgen-sensitive secretory epithelial cells in the prostate, with little effect on basal epithelial and stromal cells). PATIENTS AND METHODS: The expression of the EGF system was evaluated by enzyme-linked immunosorbent assay and immunohistochemistry in samples of prostate tissue and secretions from patients with BPH randomized for treatment with finasteride or placebo for 3 months before surgery. RESULTS: Prostate tissue expressed the EGF receptor (HER1) and HER2, and the ligands EGF, transforming growth factor alpha (TGFalpha), heparin-binding (HB) EGF, betacellulin and amphiregulin. Treatment with finasteride produced greater concentrations of amphiregulin (P < 0.05) than did placebo, did not change the level of TGFalpha, HER1 and HER2, and tended to decrease the concentration of EGF, betacellulin and HB-EGF in prostate tissue. Using immunohistochemistry, HER1 and TGFalpha were both localized to the basal epithelial cells, and there was a strong positive correlation among the tissue concentrations of HER1, HER2 and TGFalpha. Amphiregulin localized to the luminal secretory epithelium. Prostate secretions contained only EGF, which was at levels approximately 150 times higher than in prostate tissue; treatment with finasteride did not affect the concentration of EGF in prostate secretion. CONCLUSIONS: There were only minor changes in the expression of TGFalpha, HER1 and HER2 after finasteride treatment. This may represent an important system for the continuous growth and homeostasis of the androgen-independent basal epithelial cells in the prostate. PMID- 11942970 TI - Renal cell carcinoma may adapt to and overcome anti-angiogenic intervention with thalidomide. AB - OBJECTIVE: To report on the failure of thalidomide to inhibit tumour growth in an animal model of human renal cell carcinoma (RCC). MATERIALS AND METHODS: An orthotopic xenograft model of human RCC was used in which tumour cells were implanted in the left kidney of male 'severe combined immunodeficient' mice. Thalidomide was administered by intraperitoneal injection and after 34 days the mice were killed. The extent of tumour growth was compared in treated and untreated mice. Total RNA was extracted from both tumour-affected and contralateral kidneys, and analysed by reverse transcription-polymerase chain reaction for various genes implicated in angiogenesis and metastasis in RCC. RESULTS: Thalidomide failed to inhibit the growth of xenograft tumours. The expression of angiogenic genes, e.g. vascular endothelial growth factor and fibroblast growth factor type 2 (FGF-2) within normal and tumour-affected kidney tissue was not reduced by thalidomide. Intratumoral transcription of beta3 integrin, a critical component of angiogenesis, was significantly increased in response to thalidomide treatment (P < 0.01). There was also a trend to increased expression of FGF-2 and tumour necrosis factor-alpha in thalidomide-treated tumours. CONCLUSIONS: These findings suggest that RCC is capable of adapting to the inhibitory effects of thalidomide. The current uncertainty surrounding the action of thalidomide in vivo warrants caution about its use in humans. Further studies of thalidomide should be carried out in animal models, particularly to establish its safety and effectiveness as part of a combined therapeutic strategy. PMID- 11942971 TI - The beginnings of urology in Krakow: on the 70th anniversary of the founding of the Clinic of Urology. PMID- 11942972 TI - Survival in patients with rare subtypes of renal cell carcinoma. AB - OBJECTIVE: To evaluate the survival of patients with rare malignant histological subtypes of renal cancer. PATIENTS AND METHODS: The Heidelberg classification of renal cell carcinoma (RCC) divides tumours into clear cell carcinoma (CCC), papillary cancer (PC), chromophobic cancer (ChC) and collecting duct carcinoma (CDC). Sarcomatoid tumours are in a different subgroup treated as a final stage of histological progression. Between 1990 and 1997, 319 nephrectomies were undertaken because of RCC in 317 patients. In 42 patients (13%) the pathological findings showed other than CCC; in 13 PC was confirmed histologically, in nine ChC, in 11 a mixed type of CCC and sarcomatoid type, in seven a sarcomatoid tumour and in four, CDC. RESULTS: One patient of the 13 with PC and two of the nine with ChC died. The worst prognosis was in those with CDC, CCC-sarcomatoid and sarcomatoid tumours, as all these patients died. CONCLUSION: The histopathological differentiation of RCC into subtypes gives additional useful prognostic information. PMID- 11942973 TI - Assessing the vascular-stromal coefficient in patients with benign prostatic hyperplasia or prostate cancer using transrectal ultrasonography and power Doppler analysis. AB - OBJECTIVES: To assess a coefficient of the prostatic blood supply in patients with prostate cancer or benign prostatic hyperplasia (BPH), and thus improve the diagnostic and prognostic variables needed to evaluate patients with prostate cancer. PATIENTS AND METHODS: The vascular-stromal coefficient was defined as the ratio of the blood vessel surface area and the surface area of the prostate, using power Doppler functions and transrectal ultrasonography. The index was calculated for the plane of the prostatic surface from the apex to the widest points of the peripheral zone, and for the plane across the highest density of blood vessels in the transition zone. A micobubble contrast agent was used to intensify the acoustic signals from prostatic vessels. The images were reconstructed in three dimensions. In all, 61 patients with BPH and 32 with confirmed prostate cancer were examined, and compared with a control group of young men. RESULTS: In the horizontal plane crossing the apex of the prostate, the vascular-stromal coefficient was 0.0054 for the controls, 0.0072 for BPH and 0.0067 for cancer. At the level of the highest density of blood vessels the respective values were 0.0083, 0.038 and 0.029. The respective values for the first plane after using the contrast agent were 0.025, 0.092 and 0.0365, and in the second level 0.04, 0.15 and 0.14. CONCLUSION: The vascular-stromal coefficient measured at both levels was higher in patients with BPH than in those with cancer or than in the controls. The prostatic vascular system can be computer-reconstructed in three dimensions, allowing an evaluation of asymmetry and other vascular abnormalities. The contrast agent effectively increases the vascular acoustic signals from the prostate, enhancing by a factor of five the visualization of blood vessels when using the power Doppler option. PMID- 11942974 TI - Comparison of surgery alone with surgery and adjuvant radiotherapy for pT3N0 prostate cancer. AB - OBJECTIVE: To compare the outcome between patients with pT3N0 adenocarcinoma of the prostate treated with radical prostatectomy (RP) and those receiving RP followed by a planned course of postoperative radiation therapy (RT). PATIENTS AND METHODS: During a period of 22 years 622 patients with pT3N0 prostate cancer were treated in one medical centre by RP. Of these, 199 (32%) were treated with surgery alone while 423 (68%) received planned postoperative pelvic RT (median 48 Gy). Patients were selected for RT by having a higher incidence of adverse prognostic factors than those undergoing RP alone. These prognostic factors included pathological stage (P = 0.001) preoperative prostate specific antigen (PSA) level (P < 0.001) and Gleason score (P = 0.18). The patients' median age was 66 years; the median follow-up was 6.1 years for all patients, 7 years for RP + RT and 5 years for the RP-alone. RESULTS: The 5- and 10-year actuarial survival was 92% and 73%, respectively, for RP + RT patients, and nearly identical for those in the RP-alone group (P = 0.73). The 5- and 10-year disease-free survival (DFS; PSA < 0.05 ng/mL) was 69% and 51%, respectively, for the former, and 71% and 60%, respectively, for the latter group. There was no significant difference in DFS between the treatment groups by pathological stage and Gleason score (P = 0.77). Likewise, there was no significant difference in mean and median time to relapse. A preoperative PSA level of < 10 vs 10-25 vs > 25 ng/mL did not influence overall survival but a PSA of > 25 ng/mL was predictive of DFS (P = 0.02). In a multivariate analysis the Gleason score was the most important predictor for overall survival and DFS (P < 0.001), while pathological stage was predictive of clinical recurrence and DFS (P < 0.001). After controlling for pathological stage and Gleason score, RP + RT patients were predicted to recur at 92% of the rate of RP-alone patients (P = 0.65). In all, 43 (10%) patients developed a clinical recurrence in the RP + RT group, including 30 (7%) patients with distant metastases alone, 13 (3%) with local recurrence, with an additional 88 (21%) who had PSA recurrence (PSA > 0.05 ng/mL). This compared with 13 (6.5%) patients with clinical recurrence, including seven (3.5%) with local recurrence and 23 (11.6%) with PSA > 0.05 ng/mL in the RP-alone group. Postoperative RT was well tolerated and did not add to the incidence of surgical complications. CONCLUSION: We propose that postoperative RT, as described here, helped to reduce the incidence of local recurrence and improved DFS to equal that of a lower-risk group of patients treated with RP alone. A randomized comparison is needed to define the role of adjuvant RT in patients with pT3N0 disease. PMID- 11942975 TI - Correlations among prostatic-specific antigen, Gleason score, staging and grading in patients after radical prostatectomy. AB - OBJECTIVES: To evaluate the correlation between Gleason grade, T stage, tumour grade and preoperative prostate-specific antigen (PSA) level (< or > 20 ng/mL) in patients after radical prostatectomy (RP). PATIENTS AND METHODS: The serum total PSA level was measured 3 months before surgery in 87 patients who then underwent RP between 1994 and 2000; the tumour specimens were staged and graded. RESULTS: There were 57 (66%) patients with a PSA level of < 20 ng/mL; in this group only four (7%) patients had a Gleason grade of > 7 and eight (14%) had G3 tumours. Thirty (34%) patients had a PSA level of > 20 ng/mL; in this group only two (7%) patients had a Gleason grade of > 7 and 26 (87%) had G1 and G2 tumours. CONCLUSIONS: These results confirm our previous observations that the PSA level cannot be use as the only factor to indicate RP in patients with prostate cancer, but further evaluation on more patients is needed. PMID- 11942976 TI - Vascular angiographic asymmetry on three-dimensional transrectal power Doppler ultrasonography in patients with organ-confined prostate cancer. AB - OBJECTIVE: To assess whether organ-confined prostate cancer not detected on routine transrectal ultrasonography (TRUS) can be detected using three dimensional (3-D) power Doppler methods. PATIENTS AND METHODS: A preliminary trial of the use of TRUS with 3-D power Doppler image reconstruction was conducted in 116 patients being evaluated for lower urinary tract symptoms. Using a microbubble enhancing agent, an adequate horizontal plane allowed the simultaneous visualization and assessment of the symmetry of echogenicity and vascularization of the peripheral area of both prostate lobes, which is not possible in standard TRUS. The peripheral zone vascular asymmetry in the horizontal plane was also assessed retrospectively in those patients with pT2a,b prostate carcinoma. RESULTS: Stage pT2a,b prostate carcinoma was confirmed in 26 patients; in this group, two independent investigators confirmed the vascular asymmetry between the left and right peripheral zone, in the horizontal plane, in 22 patients, giving a sensitivity of 85%. CONCLUSIONS: Vascular asymmetry allows guided biopsy in lesions that are invisible using standard TRUS. Microbubble agents are an effective method of increasing the acoustic signals from small vessels of the prostate. PMID- 11942977 TI - Evaluation of the influence of systemic neoadjuvant chemotherapy on the survival of patients treated for invasive bladder cancer. AB - OBJECTIVE: To assess the influence of neoadjuvant systemic chemotherapy using a modified methotrexate, vinblastine, doxorubicin and cisplatin (MVAC) scheme in patients with invasive bladder cancer. PATIENTS AND METHODS: Two groups of patients were reviewed retrospectively; group 1 included 51 who received chemotherapy before cystectomy and group 2 included 62 who were treated only with surgery. The mean (range) duration of follow-up was 3.2 (0.25-10.25) years. The patients in group 1 were divided into two subgroups: those with tumour confined to the bladder (T1, T2 and T3a) and the remaining patients with tumour beyond the bladder (T3b, T4a,b). The chemotherapy was administered as routine MVAC, except vinblastine and methotrexate were given at 15 and 22 days during the cycle. A mean of three cycles were administered. RESULTS: The 5-year survival rate in group 1 and 2 was 66% and 58%, respectively (P > 0.3); after 8 years of follow-up the survival rates were 58% and 33%, respectively, and significantly different (P < 0.01). CONCLUSION: Systemic chemotherapy using the modified MVAC scheme in patients subsequently undergoing radical cystectomy improved the survival rate after 8 years of follow-up. PMID- 11942978 TI - Treatment of high-risk superficial bladder cancer with maintenance bacille Calmette-Guerin therapy: preliminary results. AB - OBJECTIVE: To evaluate, in a prospective study, the effects and results of maintenance therapy with bacille Calmette-Guerin (BCG) in treating patients with high-risk superficial bladder cancer. PATIENTS AND METHODS: In all, 155 patients were enrolled in a randomized study of transurethral resection alone (53) or combined with intravesical BCG (102) as a treatment for superficial bladder cancer. BCG was administered for six consecutive weeks followed by three weekly instillations in months 3, 6, 12, 18, 24, 30 and 36 after resection. Recurrence, progression, prognostic factors and side-effects were assessed and analysed. RESULTS: After a median (range) follow-up of 23 (6-42) months, 83 of the 102 patients treated with BCG (81%) were disease-free, compared with 24 of the 53 treated with resection alone (45%). There was also a significant difference in tumour progression and time to progression between the trial arms. The disease progressed in eight patients (8%) treated with BCG and in 12 (23%) of those treated by resection alone. Independent risk factors for progression were DNA ploidy status and stage. Only the completion of treatment was predictive of outcome (risk of recurrence) for patients treated with BCG. CONCLUSION: Maintenance BCG therapy was better than resection alone in reducing the incidence of recurrence and progression in patients with high-risk superficial bladder cancer. PMID- 11942979 TI - Superficial bladder cancer T1G3: the choice of treatment. PMID- 11942980 TI - Endoscopic cure of stomal leaks in continent diversion. PMID- 11942981 TI - A nephrographic dilemma: confirmation of a venous anomaly on computed tomography. PMID- 11942982 TI - Seminoma in the testes associated with haemospermia. PMID- 11942983 TI - Evaluation of the cellular tumour rejection mechanisms in the peritumoral bladder wall after bacillus Calmette-Guerin treatment. PMID- 11942984 TI - Upper and mid-ureteric stones: a prospective unrandomized comparison of retroperitoneoscopic and open ureterolithotomy. PMID- 11942986 TI - Interstitial cystitis--a time for revision of name and diagnostic criteria in the new millennium? PMID- 11942987 TI - Genomics and urology. PMID- 11942988 TI - Laparoscopic dismembered pyeloplasty: 50 consecutive cases. PMID- 11942989 TI - The self-expanding metallic ureteric stent in the long-term management of benign ureteric strictures. PMID- 11942993 TI - Intrarenal bacillus Calmette-Guerin therapy for carcinoma in situ of the upper urinary tract: long-term follow-up and natural course in cases of failure. PMID- 11942995 TI - Mistaken priorities in diabetes research. PMID- 11942996 TI - Psychological counselling in problematic diabetes: does it help? AB - BACKGROUND: In past decades clinicians have increasingly recognized the importance of psychological support for people with diabetes and their families, and many have recommended integrating psychological counselling into routine diabetes care. It is therefore important to consider whether psychological interventions in diabetes are effective in improving clinical outcomes. METHODS: This review was limited to the literature reporting on the treatment of five common psychological problems known to complicate diabetes management: depression, eating disorders, anxiety/stress, self-destructive behaviour and interpersonal/family conflicts. A literature search was undertaken using MedLine and PsychInfo, including studies published in English peer-reviewed journals between 1990 and 2001, reporting on the effects of psychological interventions in the areas mentioned. Case studies were excluded from the review. RESULTS: In line with earlier reviews, relatively little empirical research was found to substantiate the effect of psychological counselling in complicated diabetes. Most studies are uncontrolled, and involve small samples. In total only 11 randomized controlled trials were identified. Results indicate that cognitive behaviour therapy (CBT) is effective in the treatment of depression in Type 2 diabetes patients, both in reducing depressive symptoms and HbA1c. Favourable effects have been observed in pilot studies applying CBT in the field of stress management, eating disorders and self-destructive behaviour, but future research should substantiate these preliminary findings. Behaviour family therapy proved beneficial in terms of resolving family conflicts, but did not impact glycaemic control. CONCLUSIONS: Evidence to support the effect of psychological treatment in problematic diabetes is still scarce, due to limited research in this area. Suggestions are made to further develop psychotherapeutic research in diabetes care. We conclude that future research should gain from a behavioural medicine approach to diabetes, with close collaboration between diabetologists and psychologists. PMID- 11942997 TI - A chaotic model for tight diabetes control. AB - The principles of non-linear dynamics ('chaos') theory may improve our understanding of the difficulties of blood glucose control in diabetes, may lead to alternative control strategies in selected individuals, and might even enlighten the issues involved in automated glucose control for the future. Possible experimental and mathematical methods of identifying chaotic behaviour in glucose profiles are briefly discussed. Advice based on this model is suggested for those aiming at tight blood glucose control. PMID- 11942998 TI - Adherence to prescribed oral hypoglycaemic medication in a population of patients with Type 2 diabetes: a retrospective cohort study. AB - AIMS: To evaluate the patterns and predictors of adherence in all patients with Type 2 diabetes in the community receiving treatment with a single oral hypoglycaemic drug. In particular, to test the hypothesis that one tablet per day is associated with better adherence than more than one. METHODS: The study design was a retrospective cohort study set in the Tayside region of Scotland (population approx. 400 000). Participants were residents of Tayside from 1 January 1993 until 31 December 1995 with at least 12 months of prescriptions of oral hypoglycaemic drugs (OHDs). The main outcome measures were adherence indices for sulphonylureas and metformin separately, adjusting for prescribing while hospitalized. RESULTS: Of the total 2920 subjects identified, adequate adherence (> or = 90%) was found in 31% of those prescribed sulphonylureas alone (n = 1329, median adherence = 300 days per year), and in 34% of those prescribed metformin alone (n = 528, median = 302 days per year). There were significant linear trends of poorer adherence with each increase in the daily number of tablets taken (p = 0.001) and increase in co-medication (p = 0.0001) for sulphonylureas alone after adjustment for other factors. CONCLUSIONS: In the community only one in three with Type 2 diabetes had adequate adherence to OHDs. One tablet per day administration was associated with greater adherence than multiple tablets. Poor adherence is a major obstacle to the benefit of complex drug regimens in the treatment of Type 2 diabetes. PMID- 11942999 TI - Quality assurance in screening for sight-threatening diabetic retinopathy. AB - AIMS: There is a need for continuous evaluation of screening services for diabetic retinopathy against agreed performance standards. We describe a quality assurance programme implemented in Newcastle in January 1999 and report on outcomes at 18 months. METHODS: Annual retinal screening is performed using combined retinal photography and direct ophthalmoscopy in two streams. Diabetologists perform screening in the Hospital Screening Programme, which serves patients whose diabetes is managed in specialist clinics, and trained retinal screeners perform screening in the District Screening Programme, which serves patients whose diabetes is managed in the community. Reference standard examination of dilated fundoscopy with a slit-lamp and condensing lens was performed by an ophthalmologist at periodic sessions on consecutive patients attending for screening. RESULTS: Six hundred and nine (6.4%) of 9468 patients screened underwent reference standard examination. The sensitivity and specificity of detection of sight-threatening diabetic retinopathy (STDR) was 82.5% and 98%, respectively, for the Hospital Screening Programme; 85.7% and 95.7%, respectively, for the District Screening Programme; and 83.3% and 96.8% for both services combined. One hundred and ten (18.1%) of 609 patients audited were referred to ophthalmology as a result of screening, and this led to 16 patients (2.6%) receiving laser photocoagulation for STDR. Reference standard examination identified a further four patients (0.7%) who required laser photocoagulation. CONCLUSIONS: Preliminary data indicate that satisfactory performance standards are being achieved. The National Service Framework for Diabetes requires that all units institute quality assurance for retinal screening, and we report the practical implementation of this in one district. PMID- 11943000 TI - Diagnosis of diabetes mellitus and intermediate glucose abnormalities in obese patients based on ADA (1997) and WHO (1985) criteria. AB - AIM: To assess the accuracy of the 1997 ADA criteria for diagnosing diabetes mellitus and related glucose disturbances in comparison with the reference WHO 1985 criteria in obese subjects. PATIENTS AND METHODS: In 286 men and 881 women, 15-84 years of age, with obesity (body mass index (BMI) > or = 30 kg/m2), an oral glucose tolerance test (OGTT) was carried out according to WHO standard. Patients were classified into three categories of glucose tolerance using WHO 1985 (Normal Glucose Tolerance (NGT), Impaired Glucose Tolerance (IGT) and Diabetes Mellitus (DM)) and ADA (Normal Fasting Glucose (NFG), Impaired Fasting Glucose (IFG) and DM) criteria. Prevalence of each category was compared and agreement between the two classifications was assessed. The relation between fasting plasma glucose value and diabetes, as diagnosed by WHO 1985 criteria, was studied using various regression models, cumulative frequency curves, Finch method and ROC curve. RESULTS: Compared with WHO 1985, ADA criteria strongly underestimated the prevalence rate of diabetes (3.7% vs. 10.6%) and intermediate glucose abnormalities (6.0% vs. 22.4%). Agreement between the two classifications was poor (kappa = 0.23). Moreover, many patients defined as glucose-intolerant by the WHO 1985 criteria were shifted to a more favourable metabolic status by ADA criteria. Thus, ADA criteria failed to detect 69% of WHO diabetic patients and 89% with IGT were considered as normal. According to the method, cut-off value of fasting blood glucose for detecting WHO 1985-diagnosed diabetes varied widely, from 5.3 to 6.3 mmol/l and none was satisfactory because of poor sensitivity and positive predictive value. CONCLUSION: The ADA criteria do not appear to be a good substitute for those of the WHO 1985 at identifying diabetes and intermediate glucose abnormalities in an obese population. Since it appears impossible to determine a reliable cut-off value for fasting blood glucose to identify diabetic obese subjects with sufficient sensitivity, our results justify the retention of the OGTT in clinical practice or for epidemiological studies. PMID- 11943001 TI - Influence of lipoprotein(a) on restenosis after femoropopliteal percutaneous transluminal angioplasty in Type 2 diabetic patients. AB - OBJECTIVE: The influence of vascular morphology and metabolic parameters including lipoprotein(a) (Lp(a)) on restenosis after peripheral angioplasty has been compared in Type 2 diabetes (DM) vs. non-diabetic patients (ND). RESEARCH DESIGN AND METHODS: The clinical course and risk profile of 132 (54 DM vs. 78 ND) patients with peripheral arterial occlusive disease (PAD) were observed prospectively following femoropopliteal angioplasty (PTA). Clinical examination, oscillometry, ankle brachial blood pressure index (ABI) and the toe systolic blood pressure index (TSPI) were used during follow-up. Duplex sonography and reangiography were also used to verify suspected restenosis or reocclusion. RESULTS: At the time of intervention patients with DM had a lower median Lp(a) of 9 vs. 15 mg/dl (P < 0.01) in patients without diabetes. Recurrence within 1 year after PTA occurred in 25 diabetic (= 46%, Lp(a) 12 mg/dl) and 30 non-diabetic (= 38%, Lp(a) 48 mg/dl) patients. DM patients with 1 year's patency had a median Lp(a) of 7 vs. 11 mg/dl in non-diabetic patients (P < 0.05). However, 12 months after angioplasty Lp(a) correlated negatively with the ABI (r = -0.44, P < 0.01) in diabetic and in non-diabetic patients (r = -0.20, P < 0.05). The probability of recurrence after PTA continuously increased with higher levels of Lp(a) in each subgroup of patients. CONCLUSIONS: Our data indicate that Lp(a) is generally lower in those with peripheral arterial occlusive disease and Type 2 diabetes than in non-diabetic individuals. The increased risk for restenosis with rising levels of Lp(a) is set at a lower Lp(a) in diabetes and may be more harmful for diabetic patients. PMID- 11943002 TI - Evaluation of a new screening method for detecting peripheral arterial disease in a primary health care population of patients with diabetes mellitus. AB - AIMS: To compare the ability to detect peripheral arterial disease between the traditional ankle Doppler technique for measuring ankle blood pressure and a new pulse oximetric method for measuring systolic toe pressure, in an unselected primary health care population with diabetes mellitus. METHODS: The total population with the diagnosis diabetes mellitus in two primary health care districts was studied. The population was investigated by means of pulse palpation, ankle Doppler pressure, systolic toe pressure using a pulse oximetric method, arm blood pressures, neuropathy screening and laboratory tests. RESULTS: A total of 126 patients were included in the study. In these patients, 250 extremities were investigated. Systolic ankle Doppler pressure and ankle/arm pressure indices were found to be significantly higher than the pressures and indices achieved with the pulse oximetric method (158 +/- 44 vs. 117 +/- 33 mmHg, P < 0.0001, and 1.02 +/- 0.24 vs. 0.76 +/- 0.22, P < 0.0001). Thirty-one extremities with a systolic toe pressure < 80 mmHg were found. Twenty-one of these lacked a palpable pulse in the dorsal pedial or posterior tibial artery. The pulse oximetric method gave significantly more pathological indices (Doppler index < or = 0.8, pulse oximeter index < or = 0.6) (Doppler 36/250, pulse oximeter 58/250, P = 0.003). However, the Doppler method gave significantly more indices above 1.3 compared with the pulse oximetric method (33/250 vs. 2/250, P = 0.003). CONCLUSION: This study demonstrates that ankle Doppler pressure measurements overestimate peripheral arterial pressure in a typical primary health care population. In the screening situation, this new pulse oximetric toe pressure method seems to be valuable since it can be performed in out-patient clinics and handle large numbers of patients in a short time and avoid the problem of media sclerosis. PMID- 11943003 TI - Neuropsychological functioning in older people with type 2 diabetes: the effect of controlling for confounding factors. AB - AIMS AND METHODS: Neuropsychological functioning was examined in a group of 33 older (mean age 62.40 +/- 9.62 years) people with Type 2 diabetes (Group 1) and 33 non-diabetic participants matched with Group 1 on age, sex, premorbid intelligence and presence of hypertension and cardio/cerebrovascular conditions (Group 2). RESULTS: Data statistically corrected for confounding factors obtained from the diabetic group were compared with the matched control group. The results suggested small cognitive deficits in diabetic people's verbal memory and mental flexibility (Logical Memory A and SS7). No differences were seen between the two samples in simple and complex visuomotor attention, sustained complex visual attention, attention efficiency, mental double tracking, implicit memory, and self-reported memory problems. CONCLUSIONS: These findings indicate minimal cognitive impairment in relatively uncomplicated Type 2 diabetes and demonstrate the importance of control and matching for confounding factors. PMID- 11943004 TI - Post-prandial glucose excursions following four methods of bolus insulin administration in subjects with type 1 diabetes. AB - AIMS: To determine if one method of short-acting insulin bolus administration is superior to other methods in managing a meal high in carbohydrates, calories and fat. METHODS: Nine subjects receiving continuous subcutaneous insulin infusion using insulin lispro (Humalog) agreed to consume the same meal high in carbohydrates, calories and fat on four occasions 1 week apart. They received the same dose of bolus insulin on each of the four occasions randomly assigned and beginning 10 min prior to the meal as either a single bolus, two separate boluses of one-half the same total dose (the second after 90 min), the entire bolus given as a square-wave (over 2 h) or a dual-wave (70% as a bolus and 30% as a square wave over 2 h). Blood glucose levels were measured at -60 and -30 min and at zero time, and then every half-hour for 6 h using the Hemacue in the out-patient clinic. RESULTS: Changes in blood glucose values from fasting were the lowest after 90 and 120 min (P < 0.01) when the dual wave was administered. When the dual or square-wave methods of insulin administration were used, subjects had significantly lower glucose levels after 4 h in comparison with when the single or double boluses were used (P = 0.04). CONCLUSIONS: We conclude that the dual wave provided the most effective method of insulin administration for this meal. The dual- and square-wave therapies resulted in lower glucose levels 4 h after the meal in comparison with the single and double-bolus treatments. PMID- 11943005 TI - Congenital anomalies in the offspring of women with type 1, type 2 and gestational diabetes. AB - AIM: To determine the frequency of major congenital anomalies in the offspring of women with gestational diabetes (GDM), classified according to their postpartum glucose tolerance status. METHODS: A prospective study of pregnancies in women with Type 1 diabetes (n = 221), Type 2 diabetes (n = 317) and GDM (n = 1822) between 1985 and 2000 (15 years). Congenital anomalies were detected by antenatal ultrasound or postnatal examination. RESULTS: The frequency of major congenital anomalies in the offspring was 5.9% (95% confidence interval (CI) 3.2-9.8) for women with Type 1 diabetes; 4.4% (95% CI 2.4-7.3) for women with Type 2 diabetes; and 1.4% (95% CI 0.9-2.0) for women with GDM. Two hundred and thirty-seven women with GDM (13%) had diabetes diagnosed on early (6-week) postpartum glucose tolerance testing. The frequency of major congenital anomalies in their offspring was 4.6% (95% CI 2.3-8.2), compared with 0.9% (95% CI 0.5-1.5) for the remainder of the GDM group (P < 0.0001). CONCLUSIONS: GDM is not a homogeneous group with regard to the risk of major congenital anomalies. In those with diabetes on early postpartum testing, who are likely to have had unrecognized Type 2 diabetes antedating their pregnancy, the rate of major congenital anomalies is the same as for women with established Type 1 or Type 2 diabetes. In the remainder of the GDM group, the rate does not differ from the non-diabetic background rate. PMID- 11943006 TI - Association of British Clinical Diabetologists (ABCD) survey of secondary care services for diabetes in the UK, 2000. 1. Methods and major findings. AB - OBJECTIVE: To examine the provision, and variations in, secondary care diabetes services in the UK. METHODOLOGY AND PARTICIPANTS: A postal survey of all 238 identified secondary care providers of diabetes services in 2000. RESULTS: Following two reminders, a 77% response rate was achieved. Major deficiencies in core staffing levels were recorded, with 36% of services provided by only one consultant physician with an interest in diabetes. The provision of diabetes specialist nurses was less than recommended in 87% of responses, whereas podiatry and dietetic support was unavailable in 3% and 27% of responses, respectively. Diabetes registers were not present in 28%, and a co-ordinated retinopathy screening programme unavailable in 26% of responses. Key biochemical measurements were unavailable in 9% (microalbuminuria) to 18% (HDL-cholesterol) of responses. A 'Well-Resourced Service' score was devised taking account of levels of personnel, facilities and specialized clinical services. There was a significant geographical variation in this score (P < 0.001), with the lowest score (least well-resourced services) in the Eastern NHS Region of England, and the highest score in the North-west NHS Region of England. The 'Well-Resourced Service' score was also significantly lower (P < 0.05) where there were less than two whole-time consultant physicians providing diabetes services. In contrast to other aspects of service provision, availability of dieticians and a combined diabetes ophthalmology service had declined since 1990. Of 245 recorded bids for resources and service improvements for diabetes care, the success rate overall was 44%, and lowest where bids were made for dietetic and podiatry support. CONCLUSIONS: There is presently a major shortfall in provision of secondary care diabetes services throughout the UK, with evidence that there is significant regional variation and less facilities and resources where there are less than two consultants providing specialized diabetes services. On average bids for service improvements were only successful in < 50% of cases, most usually where the service was relatively better provided for. Considerable development and investment are required nationally to ensure equitable access to specialized diabetes services, a vital component in reducing adverse diabetes outcomes. PMID- 11943007 TI - Is coeliac disease more prevalent in young adults with coexisting Type 1 diabetes mellitus and autoimmune thyroid disease compared with those with Type 1 diabetes mellitus alone? AB - AIM: It is known that patients with Type 1 diabetes mellitus are more prone to develop coeliac disease and that autoimmune thyroid disease occurs more frequently in patients with coeliac disease. We therefore assessed whether coeliac disease, either known or occult, occurs more frequently in young/middle aged adults with Type 1 diabetes and coexisting autoimmune thyroid dysfunction than in adults with Type 1 diabetes alone. METHODS: The prevalence of known coeliac disease was assessed in 509 (301 males, aged 16-55 years) patients with Type 1 diabetes, 28 (5.5%) of whom had treated autoimmune thyroid disease. In a second study 38 patients with Type 1 diabetes and coexisting autoimmune thyroid disease along with 112 patients with Type 1 diabetes alone were then screened for coeliac disease using serum IgA endomysial antibodies and IgA gliadin antibodies. RESULTS: Seven of the 509 patients (1.4%) had been diagnosed with coeliac disease and two of these had later developed autoimmune thyroid disease (both hypothyroid). The subsequent screening exercise found that one of the 38 patients with both Type 1 diabetes and thyroid disease had positive endomysial antibodies on screening. However, duodenal biopsy was negative for coeliac disease. There were two patients with positive endomysial antibodies in the group of 112 patients with diabetes only. Both had duodenal biopsy but only one was consistent with coeliac disease. CONCLUSION: The prevalence of known coeliac disease in this young adult Type 1 diabetes clinic in North-west England was 7/509 (1.4%). Two of these seven patients with coeliac disease were from the group of 28 who had autoimmune thyroid disease as well. Therefore we suggest that patients with known coeliac disease and Type 1 diabetes should be screened for autoimmune thyroid disease. The second screening study then found 3/150 (2%) to have a serological marker for coeliac disease. However, patients with both Type 1 diabetes and autoimmune thyroid disease were not more likely to have occult coeliac disease compared with those with Type 1 diabetes only. PMID- 11943008 TI - Charcot neuroarthropathy of the knee in Type 1 diabetes: treatment with total knee arthroplasty. AB - BACKGROUND: Charcot neuroarthropathy is well recognized in diabetes, although it typically affects the joints of the forefoot and ankle. Neuroarthropathy affecting the knee in diabetes is extremely rare. The surgical options for treatment of Charcot neuroarthropathy remain poorly defined, particularly the use of arthroplasty with knee involvement. METHODS: We describe a case of neuroarthropathy of the knee in a patient with Type 1 diabetes. We also describe the successful management of the disorder with total knee arthroplasty-only the third such description. RESULTS: The case illustrates some of the typical radiological features of this uncommon condition, which may aid early diagnosis and limit morbidity. PMID- 11943010 TI - Height and gestational diabetes mellitus. PMID- 11943011 TI - Handling of blood samples for diagnosis of diabetes mellitus. PMID- 11943012 TI - The threshold value for insulin resistance on homeostasis model assessment of insulin sensitivity. PMID- 11943013 TI - Pioglitazone-induced hepatic injury in a patient previously receiving troglitazone with success. PMID- 11943015 TI - Recent advances in immunohistochemistry in gynaecological pathology. AB - Recent years have witnessed significant developments in the use of immunohistochemistry in diagnostic gynaecological pathology. This review details the most significant of these. In ovarian pathology, differential cytokeratin staining (CK7 and 20) assists in distinguishing between a primary ovarian adenocarcinoma and a metastatic adenocarcinoma, especially of colorectal origin. The development of markers characteristic of ovarian sex cord-stromal tumours (especially alpha-inhibin) facilitates diagnosis of these neoplasms which is often difficult by morphology alone due to the wide differential diagnosis. In the uterus, the distinction between a primary endometrial and endocervical adenocarcinoma may be facilitated by use of a small panel of antibodies, including CEA, ER and vimentin. Newly developed antibodies such as CD10 and h caldesmon may be of use in the diagnosis of uterine mesenchymal lesions, especially in the distinction between endometrial stromal and smooth muscle lesions. Proliferation markers, such as MIB1, are of value in the cervix in the diagnosis of preinvasive squamous and glandular lesions. Recent studies have shown that cervical adenoma malignum exhibits a gastric phenotype. Advances have also been made in trophoblastic disease with the development of antibodies reactive against trophoblast such as alpha-inhibin, mel-Cam and p57. A newly developed monoclonal antibody HMGIC which is expressed in vulvovaginal aggressive angiomyxoma may prove to be of value in the often difficult distinction of this lesion from its histological mimics. PMID- 11943016 TI - Clinicopathological significance of the 'keloid-like' collagen and myxoid stroma in advanced rectal cancer. AB - AIM: To establish the histological categorization of fibrotic stroma which reflects the biological behaviour of advanced rectal cancer. METHODS AND RESULTS: Six hundred and twenty-seven surgically resected cases of advanced rectal carcinoma were examined. We histologically categorized fibrotic stroma in the invasive frontal region into three groups: type A, multiple fine and mature fibres were stratified into layers; type B, broad bands of eosinophilic hyalinized collagen ('keloid-like' collagen) were intermingled; type C, myxoid stroma. Type A stroma was observed in 63% of patients, type B stroma in 25%, type C stroma in 12%. The incidence of type A stroma decreased in accordance with Dukes stage (98% in Dukes A; 73% in B; 41% in C1; 29% in C2) and conversely, there was an increase of C type (0% in Dukes A; 4% in B; 20% in C1; 54% in C2). Stroma type had a significant correlation with long-term survival (80% of 5-year survival in type A stroma; 54% in type B; 26% in type C). Based on multivariate analysis, it was found that the stromal pattern had independent prognostic value, together with nodal involvement, growth pattern, and lymphocyte infiltration. CONCLUSIONS: Tumour fibrotic stroma may play an important role as a regulator of neoplastic behaviour. Pathological categorization of the fibrotic stroma is helpful for predicting the prognostic outcome of patients with rectal carcinoma. PMID- 11943017 TI - Microscopic colitis with giant cells. AB - AIMS: Collagenous colitis and lymphocytic colitis are the two types of microscopic colitis with specific morphological features. In this report we describe a new histopathological subtype of microscopic colitis. METHODS AND RESULTS: Colonoscopy in four patients with chronic watery diarrhoea showed no macroscopic abnormalities. The random biopsies from the colon showed subepithelial multinucleated giant cells in combination with the features of collagenous colitis in three patients and lymphocytic colitis in one patient. These multinucleated giant cells were positive for CD68. The density of macrophages was highest in the most superficial part of the lamina propria. In one patient, a previous biopsy showed features consistent with collagenous colitis without multinucleated giant cells. Treatment with budesonide led to the disappearance of diarrhoea in all four patients. CONCLUSIONS: The clinical and histopathological features of the four presented patients indicate that there exists a histopathological subtype of microscopic colitis characterized by the presence of subepithelial multinucleated giant cells, which probably arise from fusion of subepithelial macrophages. Analysis of more patients with this histopathological subtype of microscopic colitis is necessary to determine whether they also form a clinically distinct group. PMID- 11943018 TI - Enhanced expression of decay-accelerating factor and CD59/homologous restriction factor 20 in intestinal metaplasia, gastric adenomas and intestinal-type gastric carcinomas but not in diffuse-type carcinomas. AB - AIMS: Variable expression of the complement regulatory proteins, decay accelerating factor, CD59/homologous restriction factor 20 (HRF20) and membrane cofactor protein has been shown in human gastrointestinal malignancies, but their expression in gastric cancer has not been fully described. Thus, we immunohistochemically defined the distribution of these proteins in human normal gastric mucosa, intestinal metaplasia, adenomas and gastric cancers. METHODS AND RESULTS: Gastric tissues were obtained by endoscopic biopsy or surgical resection and stained with mouse monoclonal antibodies to decay-accelerating factor, CD59/HRF20, and membrane cofactor protein. In the normal gastric mucosa, membrane cofactor protein was diffusely stained on the basolateral surface of epithelial cells, whereas the expression of decay-accelerating factor and CD59/HRF20 was inconspicuous. In intestinal metaplasia, adenoma and intestinal-type gastric carcinoma cells, decay-accelerating factor and HRF20 were intensely stained on the apical surface; membrane cofactor protein retained its location on the basolateral surface. In diffuse-type gastric carcinomas, the expression of decay accelerating factor, CD59/HRF20 was lost, but membrane cofactor protein was present on the tumour cell surface. CONCLUSIONS: These findings suggest that membrane cofactor protein plays a primary role in the regulation of complement activation in normal and neoplastic gastric cells and that the expression pattern of the complement regulatory proteins is closely related to gastric carcinoma development. PMID- 11943019 TI - Malignant deciduoid mesothelioma of the pleura: report of two cases with long survival. AB - AIMS: To present two rare cases of malignant mesotheliomas with deciduoid features arising in the pleura, both with long survival. METHODS AND RESULTS: These two cases of deciduoid mesotheliomas were observed in adult patients (one 73-year-old male and one 23-year-old female). Only the male had a history of occupational asbestos exposure, whereas the woman had a history of familial mesothelioma. A deciduoid morphology was predominant and focal areas with tubular papillary features were noted. The tumour cells were positive for cytokeratins, HMBE-1, calretinin, EMA and mitochondrion antibodies. The follow-up data did not suggest a particularly poor prognosis; the mean survival observed was 23 months (17 and 39 months, respectively). CONCLUSIONS: This deciduoid mesothelioma histological subtype does not appear to represent an unfavourable prognostic category. PMID- 11943020 TI - Clinicopathological prognostic factors in soft tissue leiomyosarcoma: a multivariate analysis. AB - AIMS: Prognostic factors affecting survival in cases of leiomyosarcoma of soft parts were investigated in this study. METHODS AND RESULTS: A retrospective study of 267 patients was carried out. This group comprised 142 females (53%) and 125 males (47%), whose ages ranged from 7 to 95 years (median 58 years). One hundred and five cases were superficially situated (arising from the skin or subcutis), while the remaining 162 cases were deeply situated (subfacial). Nineteen were cases of pleomorphic leiomyosarcoma where the diagnosis had been amended from malignant fibrous histiocytoma to leiomyosarcoma whilst under review. Of the 167 patients with follow-up data, 83 died of leiomyosarcoma. In univariate analysis, depth, tumour size (>or=50 mm), mitotic rate of >20 per 10 high-power fields (HPF), tumour necrosis of >50% and a high stage according to the most recent American Joint Committee on Cancer (AJCC) staging for soft tissue sarcoma were found to lessen significantly the rate of survival (log rank test; P < 0.05). However, in multivariate analysis (Cox's proportional hazards model), tumour size and high AJCC stage were the only factors that were correlated independently with decreased survival. CONCLUSIONS: This study indicates that the most reliable prognostic parameters are tumour size and AJCC stage in leiomyosarcoma. PMID- 11943021 TI - Nuclear to cytoplasmic compartment shift of the p33ING1b tumour suppressor protein is associated with malignancy in melanocytic lesions. AB - AIMS: Cutaneous malignant melanoma is an unpredictable neoplasm. Studies of cell cycle and proliferation-associated proteins may help in the understanding of the genesis of melanomas. The tumour suppressor gene TP53 has been shown to be involved in melanomas. However, the incidence of TP53 malfunction in cutaneous melanoma is unclear, and other regulators of cell cycle control are likely to be involved in both the development and progression of melanocytic neoplasia. A candidate is the ING1 gene, which co-operates with TP53 in growth suppression and apoptosis. Thus loss of ING1 function may have similar consequences to loss of TP53 function and may contribute to tumorigenesis. Therefore we have studied the expression of p33ING1b protein in cutaneous melanocytic neoplasia. METHODS AND RESULTS: Sixty-seven melanocytic lesions were studied by immunohistochemistry for the expression of p33ING1b. In our series there was loss of nuclear p33ING1b expression in invasive malignant melanoma compared with normal cutaneous melanocytes or the melanocytes of benign melanocytic naevi. This was associated with an enhancement of cytoplasmic p33ING1b expression which was particularly prominent in invasive malignant melanoma. CONCLUSIONS: Cytoplasmic immunostaining for p33ING1b using MAb GN2 is strongly associated with 'activated' melanocytic lesions; therefore it is possible that this MAb could be of value in diagnostic practice. Furthermore, the reduction in p33ING1b expression and perhaps translocation from the nucleus to the cytoplasm may play a central role in the development and progression of melanomas. PMID- 11943022 TI - Expression of respiratory mucins in fatal status asthmaticus and mild asthma. AB - AIMS: The airways of patients with asthma are characterized by chronic inflammatory changes comprising mainly T-cells and eosinophils, and airway remodelling with goblet cell metaplasia and submucosal gland hyperplasia. Mucus hypersecretion is often a marked feature, particularly in status asthmaticus. The matrix of airway sputum consists of high molecular glycoproteins and mucins. In this study, the expression and distribution of the major gel-forming mucins MUC5AC and MUC5B were studied in fatal status asthmaticus tissues and bronchial biopsies of mild asthmatic patients. The effect of inhaled corticosteroids on the expression of these mucins was also investigated. METHODS AND RESULTS: Polyclonal antibodies specific for MUC5AC and MUC5B, and a monoclonal antibody for MUC5B were used to stain lung tissues and airway mucosal biopsies obtained from patients who died of status asthmaticus (n=5) and from mild asthmatics (n=4), respectively. Immunohistochemistry for MUC5AC revealed abundant staining of goblet cells situated in the epithelial surface lining and glandular ducts of tissues from patients with fatal asthma. MUC5B immunoreactivity was restricted to mucous cells of submucosal glands and to epithelial cells. In mild asthmatics, large amounts of MUC5B, but not MUC5AC, positive extracellular mucus was found in the airway lumen as plugs, adjacent to the epithelial lining and in the necks of glandular secretory ducts of mild asthmatics. The distribution of MUC5AC and MUC5B in bronchial biopsies of mild asthmatics was similar before and after inhaled steroid treatment. CONCLUSIONS: The expression of MUC5AC and MUC5B shares a similar distribution to normal airways in different states of asthma. The distribution is not affected by topical corticosteroid therapy. PMID- 11943023 TI - 'Microscopic colitis': classification and terminology. PMID- 11943024 TI - How should we grade CIN? AB - How should we grade CIN? The grading of cervical intraepithelial neoplasia (CIN) is problematic. CIN represents a morphological continuum, but biopsies displaying this lesion are classified into two (e.g. Bethesda) or three grade categories, sometimes with poor reproducibility. There are also difficulties in reliably distinguishing low-grade CIN from its reactive simulants. Because of problems with inter- and intra-observer disagreement in the grading of CIN and the diagnosis of low-grade lesions, three expert contributions were commissioned to address the question 'how should we grade CIN?'. PMID- 11943027 TI - Cyclin E and p27/Kip1: two proteins on a see-saw or another false dawn? PMID- 11943028 TI - Ileal subepithelial collagen thickening in an HIV+ patient presenting with diarrhoea with no evidence of collagenous colitis. PMID- 11943029 TI - Gynandroblastoma of the testis in an infant: a morphological, immunohistochemical and in-situ hybridization report. PMID- 11943030 TI - Mammary fibromatosis with extensive necrosis. PMID- 11943033 TI - Alendronate reduces the risk of multiple symptomatic fractures: results from the fracture intervention trial. AB - OBJECTIVES: To evaluate the effect of alendronate on the occurrence rate of multiple svmptomatic fractures and on the risk of multiple symptomatic fractures (likelihood of having more than one fracture diagnosed because of the symptoms the fractures caused over the study period) among women with osteoporosis. DESIGN: Primary analysis of data from a randomized, placebo-controlled, double blind trial. SETTING: Eleven community-based clinical research centers. PARTICIPANTS: Subset of women enrolled in the Fracture Intervention Trial: aged 55 to 81 and having at least one morphometric vertebral fracture at baseline (n=2,027) or having no vertebral fracture but meeting prevailing World Health Organization bone mineral density criteria for osteoporosis (T-score < or =2.5 at the femoral neck)(n = 1,631). INTERVENTION: All participants reporting calcium intake of 1,000 mg/day or less received a supplement of 500 mg calcium and 250 IU cholecalciferol. Participants were randomly assigned to placebo or alendronate sodium (5 mg/day for 2 years and 10 mg/day for the remainder of the study). Average total follow-up was 4.3 years. MEASUREMENTS: Symptomatic fractures were diagnosed by personal physicians and confirmed by review of radiological data by an expert committee blinded to treatment assignments. RESULTS: Eighty-six of 1,817 women receiving placebo experienced multiple symptomatic fractures during the follow-up period, compared with 51 of 1,841 receiving alendronate. Reduction of risk for multiple symptomatic fractures combined was 42% (relative risk (RR) = 0.58, 95% confidence interval (CI) = 0.41, 0.81) and for multiple symptomatic vertebral fractures was 84% (RR = 0.16,95% Cl = 0.05, 0.42). Cumulative incidence curves showed divergence after as little as 3 months of treatment, with a statistically significant (P = .044) reduction at 6 months for multiple symptomatic vertebral fractures. When all fractures over the follow-up period were included, the occurrence rates of all symptomatic fractures and symptomatic vertebral fractures were 34% and 63% lower, respectively, with alendronate than with placebo. These reductions were sustained during the follow-up period. All reductions in risk were consistent across predefined subgroups: age (<75 vs > or =75), morphometric vertebral fracture(present vs absent), prior clinical fracture since age 45 (yes vs no), and whether the subject had fallen in the 12 months before randomization. CONCLUSIONS: These data demonstrate that treatment with alendronate reduces the risk of multiple symptomatic fractures during a treatment period averaging 4.3 years. The reductions were consistent across prespecified sub-groups. This effect is evident early in treatment and is sustained. PMID- 11943034 TI - Glucose, blood pressure, and lipid control in older people with and without diabetes mellitus: the Cardiovascular Health Study. AB - OBJECTIVES: To determine the prevalence of cardiovascular risk-factor treatment and control in older adults with normal fasting glucose, impaired fasting glucose, and diabetes mellitus and whether those with diabetes mellitus had better risk factor control than older adults with normal fasting glucose. DESIGN: Secondary analysis of data from population-based, prospective cohort study of risk factors for cardio-vascular and cerebrovascular disease in older people (Cardiovascular Health Study). SETTING: Community-based. PARTICIPANTS: Community dwelling adults aged 65 and older. MEASUREMENTS: Fasting plasma glucose, serum cholesterol and its subfractions, systolic and diastolic blood pressures, and body mass index. RESULTS: There were 579 (18%) cohort members with diabetes mellitus (77% receiving antidiabetic medication, 23% with fasting glucose > or =126 mg/dL and no treatment), 213 (6%) with impaired fasting glucose, and 2,582 (77%)with normal fasting glucose. Of diabetic participants, 12% had recommended fasting glucose levels of less than 110 mg/dL. Of participants with hypertension, a larger proportion of diabetic participants than nondiabetic participants (89% versus 75%, P < .01) was treated with antihypertensive agents, but a smaller proportion of diabetic participants had recommended blood pressure levels of 129/85 mmHg or lower than nondiabetic participants had recommended blood pressure levels of 139/89 mmHg or lower (27% vs 48%, P < .01). Diabetic dyslipidemic participants were treated less often with lipid-lowering therapy (26% versus 55%, P < .01) and achieved recommended low-density lipoprotein goals less often (8%versus 54%, P < .01) than nondiabetic dyslipidemic participants. CONCLUSIONS: Overall, treatment and control of cardiovascular risk factors were suboptimal in this older population, especially among those with diabetes mellitus. Optimizing risk-factor control can improve health outcomes in older adults with and without diabetes mellitus. PMID- 11943035 TI - Relationship of race/ethnicity and blood pressure to change in cognitive function. AB - OBJECTIVES: To determine whether there are racial/ethnic differences regarding the relationship of level of blood pressure to change in cognitive function in older people. DESIGN: Longitudinal data 1986 to 1989 on representative, older, community-residing African Americans and whites. Blood pressure levels were assessed and a brief screen of cognitive functioning, the Short Portable Mental Status Questionnaire (SPMSQ), was performed at baseline and 3 years later. SETTING: Five contiguous counties in the Piedmont area of North Carolina. PARTICIPANTS: African-American (n = 2,260) and white(n = 1,876) participants in the Duke Established Populations for Epidemiologic Studies of the Elderly, aged 65 to 105 at baseline. MEASUREMENTS: The outcome measure was change in SPMSQ score over 3 years. Covariates included age; education; gender; self-reported diabetes mellitus, stroke, heart attack, current smoking, and depressive symptomatology;and use of antihypertensive medication. The primary independent variable was measured blood pressure. RESULTS: In unadjusted analyses, a statistically significant U-shaped relationship was found between systolic (but not diastolic) blood pressure levels and change in SPMSQ score over a 3-year period for older white men and women. No such relationships were found between these blood pressure measurements and change in SPMSQ score in older African Americans. These findings remained after adjustment for initial SPMSQ score, demographic characteristics, and use of antihypertensive medication. There were no significant interactions between race and blood pressure on change in cognitive function. CONCLUSION: Decline in cognitive function was associated with extremes of systolic blood pressure in older white people. Although a similar but muted nonsignificant association was found in older African Americans, the curves for the two groups were not significantly different. Further studies in older African Americans are needed. PMID- 11943036 TI - Oral care reduces pneumonia in older patients in nursing homes. AB - OBJECTIVES: Aspiration of oral secretions and their bacteria is increasingly being recognized as an important factor in pneumonia. We investigated whether oral care lowers the frequency of pneumonia in institutionalized older people. DESIGN: Survey. SETTING: Eleven nursing homes in Japan. PARTICIPANTS: Four hundred seventeen patients randomly assigned to an oral care group or a no oral care group. INTERVENTION: Nurses or caregivers cleaned the patients' teeth by toothbrush after each meal. Swabbing with povidone iodine was additionally used in some cases. Dentists or dental hygienists provided professional care once a week. MEASUREMENTS: Pneumonia, febrile days, death from pneumonia, activities of daily living, and cognitive functions. RESULTS: During follow-up, pneumonia, febrile days, and death from pneumonia decreased significantly in patients with oral care. Oral care was beneficial in edentate and dentate patients. Activities of daily living and cognitive functions showed a tendency to improve with oral care. CONCLUSION: We suggest that oral care may be useful in preventing pneumonia in older patients in nursing homes. PMID- 11943037 TI - Community-acquired pneumonia in older veterans: does the pneumonia prognosis index help? AB - OBJECTIVES: Mortality rates from pneumonia increase steadily with age. Recently, a disease severity model (the Pneumonia Prognosis Index (PPI)) has been developed to predict mortality from community-acquired pneumonia (CAP). PPI ranks severity of pneumonia from 1 to 5, with 5 being most severe. This retrospective study utilizes the PPI to address the prognosis of CAP in older adults. DESIGN: Retrospective review of medical charts. SETTING: Department of Veterans Affairs Medical Center. PARTICIPANTS: All adults aged 60 and older admitted to a Veterans Affairs Medical Center with CAP between January 1 and December 31, 1998. MEASUREMENTS: PPI was calculated using subjects' demographics, comorbidities, presenting symptoms, and laboratory measurements. RESULTS: Eighty-two patients aged 60 and older were admitted with 101 episodes of CAP. The mean age +/- standard deviation was 72 +/- 9. Seventy-four episodes were admitted from the emergency room, 20 from another hospital, seven from nursing homes, and five from outpatient clinics. Mean length of stay was 7.1 +/- 6 days. Comorbid conditions included coronary artery disease, diabetes mellitus, congestive heart failure, neoplasm, stroke, and renal failure. Most episodes fell into higher PPI classes, with 20% in Class 3, 46% in Class 4, and 32% in Class 5. PPI score was significantly related to length of stay (P < .001), intensity of care (P < .0001), and presence of complications (P <.001). Mortality was 14% at 30 days: 0.5% in Class 3,10.8% in Class 4, and 25% in Class 5. CONCLUSION: The PPI was effective in identifying older adults with CAP who were at risk of a poor outcome, but the practical utility of this index remains to be determined.Further prospective studies are required to elucidate the importance of comorbidities, severity at presentation, and premorbid functional status on clinical and functional outcomes of CAP in older adults. PMID- 11943038 TI - Severe dementia and adverse outcomes of nursing home-acquired pneumonia: evidence for mediation by functional and pathophysiological decline. AB - OBJECTIVES: To assess whether the severity of dementia is related to unfavorable outcomes of nursing home-acquired pneumonia and how this relationship is mediated. DESIGN: Prospective cohort study. SETTING: Psychogeriatric wards of 61 nursing homes in the Netherlands. MEASUREMENTS: Dementia severity and the possible mediators swallowing disturbance, aspiration, insufficient food intake, weight loss, and dehydration were measured and related to the following outcomes: death (rate), cure rate, and increase in discomfort at the onset of pneumonia. PARTICIPANTS: Demented patients (n = 374) treated with antibiotics for pneumonia. RESULTS: Dementia severity was independently related to death rate within the first week after pneumonia (hazard rate ratio = 3.0 for the most severely demented quartile versus the least demented quartile, 95% confidence interval (CI) = 1.1-8.3) and to 3-month mortality (odds ratio (OR) 2.5, 95% CI = 1.1-5.4). The latter relation was in part mediated by aspiration and weight loss (OR dementia severity adjusted for these mediators declined from 2.5 to 1.9, 95% Cl = 0.8-4.3). Dementia severity was not related to cure rate within 2 weeks nor to an in-crease in discomfort after 3 days compared with before the pneumonia. CONCLUSION: The functional and pathophysiological consequences of progressive dementia account in part for increased 3-month mortality after pneumonia. Mid term mortality is expected to be high only in the most severely demented patients and in less severely demented patients who aspirated or who lost weight. Implications for end-of-life decision-making and effectiveness of preventive and curative interventions are discussed. PMID- 11943039 TI - Clinical features and functional outcome of intracerebral hemorrhage in patients aged 85 and older. AB - OBJECTIVES: The importance of studies of older stroke victims is growing because of the changes in the age structure of Western populations. We determined demographic characteristics, clinical features, neuroimaging data, and outcome of acute spontaneous intracerebral hemorrhage occurring in people aged 85 and older. DESIGN: Prospective collection of data from a hospital-based registry. SETTING: Sagrat Cor Hospital of Barcelona (an acute care, 350-bed teaching hospital serving a population of approximately 250,000). PARTICIPANTS: Between January 1986 and December 1995, data were collected on 2,000 stroke patients admitted consecutively to the department of neurology (25 beds and an acute stroke unit). MEASUREMENTS: For the purpose of this study, all cases of intracerebral hemorrhage were selected (n = 229). Demographic data, clinical features, neuroimaging findings, and outcome variables (in-hospital mortality and medical complications developed during hospitalization)in patients aged 85 and older (n = 28) were compared with patients who were younger than age 85 (n = 201). Distinctive clinical features of intracerebral hemorrhage in very old people were assessed by multivariate analysis. RESULTS: Intracerebral hemorrhage was diagnosed in 11% of subjects aged 85 and older included in the stroke registry and accounted for 12% of all cases of intracerebral hemorrhage. Very old people with intracerebral hemorrhage showed a significantly higher frequency of female gender (75% vs 32%, P < .005), altered consciousness (64% vs 43%, P < .05), multiple topographic involvement (29% vs 13%, P < .03), undetermined etiology of bleeding (50% vs 27%, P < .02), moderate or severe neurological deficit at hospital discharge (89% vs 58%, P < .005), and in-hospital mortality (50% vs 27%, P < .01) than younger people. After multivariate analysis, female sex (odds ratio (OR) = 3.2,95% confidence interval (CI) - 1.27-7.99) and moderate or severe neurological deficit at hospital discharge (OR =4.75, 95% Cl = 1.36-16.55) were independent clinical factors associated with intracerebral hemorrhage in very old people. CONCLUSIONS: Patients aged 85 and older with intra-cerebral hemorrhage showed some peculiar clinical features and poorer outcome, including higher in hospital mortality and moderate or severe neurological deficit at hospital discharge, than younger patients with intracerebral hemorrhage. PMID- 11943040 TI - Rapid resting heart rate: a simple and powerful predictor of osteoporotic fractures and mortality in older women. AB - OBJECTIVES: To determine whether resting heart rate is associated with several types of osteoporotic fractures, mortality, and cause-specific mortality in older women. DESIGN: A prospective cohort study. SETTING: Four communities across the United States. PARTICIPANTS: We prospectively assessed resting pulse rate in 9,702 women aged 65 and older enrolled in the Study of Osteoporotic Fractures. MEASUREMENTS: Resting pulse was measured in the supine position. Hip, humerus, pelvis, rib, ankle, and wrist fractures were identified by self-report and validated by radiographs. Incident vertebral fractures were assessed by quantitative morphometry. Cause-specific mortality was assessed from death certificates and hospital discharge summaries. RESULTS: Women with resting heart rates of greater than 80 beats per minute (bpm) (n = 1,140 (12%)) had an adjusted 1.6-fold (95% confidence interval (CI) = 1.2-2.0) increased risk of either a hip, pelvis, or rib fracture and a 1.9-fold (95% CI = 1.4-2.5) increased risk of vertebral fracture. A heart rate of 80 bpm or greater was also associated with 1.4-fold (95% Cl = 1.2-1.5) increased all-cause mortality and 1.5-fold (95% CI = 1.2-2.1) increased coronary heart disease mortality. Investigating resting heart rate as a continuous variable, we detected a general pattern of increasing risks with higher heart rate that could not be explained by age, weight, poor health, physical activity, hyperthyroidism, or smoking. CONCLUSIONS: Older women with resting heart rates of 80 bpm or more have an increased risk of several osteoporotic fractures and of mortality that is not explained by other risk factors. Heart rate may be a simple tool for assessing risk of fracture and of death from coronary heart disease in older women. PMID- 11943041 TI - The relationship between leg power and physical performance in mobility-limited older people. AB - OBJECTIVES: The purpose of this study was to assess the influence of leg power and leg strength on the physical performance of community-dwelling mobility limited older people. DESIGN: Cross-sectional analysis of baseline data from a 12 week randomized controlled exercise-intervention study. SETTING: Exercise laboratory within the Department of Health Science of an urban university. PARTICIPANTS: Forty-five community-dwelling mobility-limited people (34 women, 11 men), aged 65 to 83. MEASUREMENTS: Health status, depression, cognition, physical activity, and falls efficacy; physiological measures of lower extremity strength and power; and measures of physical performance. RESULTS: Through bivariate analyses, leg power was significantly associated with physical performance as measured by stair-climb time, chair-stand time, tandem gait, habitual gait, maximal gait, and the short physical performance battery describing between 12% and 45% of the variance (R2). Although leg power and leg strength were greatly correlated (r = .89) in a comparison of bivariate analyses of strength or power with physical performance, leg power modeled up to 8% more of the variance for five of six physical performance measures. Despite limitations in sample size, it appeared that, through quadratic modeling,the influence of leg power on physical performance was curvilinear. Using separate multivariate analyses, partial R2 values for leg power and leg strength were compared, demonstrating that leg power accounted for 2% to 8% more of the variance with all measures of physical performance. CONCLUSION: Leg power is an important factor influencing the physical performance of mobility-limited older people. Although related to strength, it is a separate attribute that may exert a greater influence on physical performance. These findings have important implications for clinicians practicing geriatric rehabilitation. PMID- 11943042 TI - The relationship between lower body strength and obstructed gait in community dwelling older adults. AB - OBJECTIVES: To determine the relationship between lower body strength of community-dwelling older adults and the time to negotiate obstructed gait tasks. DESIGN: A correlational study. SETTING: The Biomechanics Laboratory, Deakin University, Australia. PARTICIPANTS: Twenty-nine women and 16 men aged 62 to 88 were recruited using advertisements placed in local newspapers. The participants were independent community dwellers, healthy and functionally mobile. MEASUREMENTS: Maximal isometric strength of the knee extensors and dynamic strength of the hip extensors, hip flexors, hip adductors, hip abductors, knee extensors, knee flexors, and ankle plantar flexors were assessed. The times to negotiate four obstructed gait tasks at three progressively challenging levels on an obstacle course and to complete the course were recorded. The relationship between strength and the crossing times was explored using linear regression models. RESULTS: Significant associations between the seven strength measures and the times to negotiate each gait task and to walk the entire course at each level were obtained (r = -0.38 to -0.55; P < .05). In addition, the percentage of the variance explained by strength (R(2)), consistently increased as a function of the progressively challenging level. This increase was particularly marked for the stepping over task (R(2) = 19.3%, 25.0%, and 27.2%, for levels 1, 2, and 3, respectively) and the raised surface condition (R2 = 17.1%, 21.1%, and 30.8%, for levels 1, 2, and 3, respectively). CONCLUSION: The findings of the study showed that strength is a critical requirement for obstructed locomotion. That the magnitude of the association increased as a function of the challenging levels suggests that intervention programs aimed at improving strength would potentially be effective in helping community-dwelling older adults negotiate environmental gait challenges. PMID- 11943043 TI - Development and validation of a risk-adjustment index for older patients: the high-risk diagnoses for the elderly scale. AB - OBJECTIVES: The purpose of this study was to develop and validate a risk adjustment index for 1-year mortality specific to older people, based on administrative discharge diagnoses. DESIGN: Two prospective cohort studies, in tandem. The index developed in the initial cohort was subsequently validated in a separate cohort. SETTING: General medicine service of a university teaching hospital. PARTICIPANTS: For the development cohort, 524 hospitalized general medical patients aged 70 and older. For the validation cohort, 852 comparable patients. MEASUREMENTS: Administrative diagnosis data were used to construct the proposed index and several other widely used indices (Deyo-adapted Charlson; Acute Physiology, Age, Chronic Health Evaluation III conditions; total number of diagnoses; All Patient Refined Diagnosis Related Groups; and Disease Staging). We used receiver operating characteristic curve analysis and Cox proportional hazards modeling to compare our proposed index with the other indices with respect to predictive accuracy and strength of association with 1-year mortality. RESULTS: The High-Risk Diagnoses for the Elderly Scale was developed using 10 high-risk medical diagnoses. Individual condition weights, based on the magnitude of 1-year mortality risk, ranged from 1 (pneumonia, diabetes mellitus with end organ damage) to 6 (lymphoma/leukemia); possible index scores ranged from 0 to 27. Mortality rates for patients categorized into four risk groups based on the index were 9.5%, 31.8%, 46.4%, and 73.6% in the development cohort (C statistic = 0.76), and 9.9%, 24.3%, 33.6%, and 50.8% in the validation subjects (C statistic = 0.68). The new index was a stronger predictor of mortality than several widely used measures. CONCLUSION: The High-Risk Diagnoses for the Elderly Scale, based on readily available administrative data,is a simple, accurate system for prediction of 1-year mortality in older hospitalized patients that demonstrated generalizability to an independent sample. Future studies are needed to test this index in other settings and populations. PMID- 11943044 TI - Central auditory dysfunction may precede the onset of clinical dementia in people with probable Alzheimer's disease. AB - OBJECTIVES: To document the prognostic significance of a central auditory speech processing deficit for the subsequent onset of probable Alzheimer's disease. DESIGN: Prospective cohort study. SETTING: Framingham Heart Study. PARTICIPANTS: Seven hundred forty dementia-free volunteers from the Framingham Heart Study cohort with symmetric hearing thresholds at biennial examination 15 (1983-1985). MEASUREMENTS: The diagnosis of probable Alzheimer's disease was made prospectively using the National Institute of Neurological and Communicative Disorders and Stroke-Alzheimer's Disease Related Disorder Association criteria. A central auditory speech-processing deficit was defined as a score of 50% of less correct on the Synthetic Sentence Identification with Ipsilateral Competing Message test in at least one ear with normal word recognition ability in both ears. Cox proportional hazards regression assessed the relationship between a central auditory speech-processing deficit and the age at diagnosis of probable Alzheimer's disease. RESULTS: Forty subjects (5.4%) received a diagnosis of probable Alzheimer's disease during an average of 8.4 years (range 3-12) of follow-up; seven (17.5%) of these had a central auditory speech-processing deficit. The presence of a central auditory speech-processing deficit had an age adjusted risk ratio for probable Alzheimer's disease of 10.8 (95% CI = 4.6-25.2), and the estimated risk ratio adjusted for age, gender, education level, apolipoprotein allele E4 presence, and hearing level was 23.3 (95% CI =6.6-82.7). A central auditory speech-processing deficit had a positive predictive value for subsequent probable Alzheimer's disease of 47% but the sensitivity was only 17.5%. CONCLUSION: Central auditory speech-processing deficits may be an early manifestation of probable Alzheimer's disease and may precede the onset of dementia diagnosis by many years. PMID- 11943045 TI - Urinary incontinence and psychological distress in community-dwelling older adults. AB - OBJECTIVES: To examine the relationship between urinary incontinence (UI) and psychological distress in older adults. We hypothesized that persons with UI associated with condition-specific functional loss would be most likely to report psychological distress. DESIGN: A population-based longitudinal survey. SETTING: Continuing participants in a study of community-dwelling adults who were initially living in East Baltimore in 1981. PARTICIPANTS: Persons aged 50 and older (n=781) at follow-up interviews conducted between 1993 and 1996 for whom complete data were available. MEASUREMENTS: Participants were classified as incontinent if they reported any uncontrolled urine loss within the 12 months preceding the 13-year follow-up interview. Condition-specific functional loss secondary to UI was further assessed based on a series of questions relating directly to participants' inability to engage in certain activities due to their UI. Psychological distress was assessed using the General Health Questionnaire (GHQ) at interviews in 1981 and at the 13-year follow-up. RESULTS: Persons with UI were more likely to experience psychological distress as measured by the GHQ than were persons without UI (unadjusted odds ratio (OR)=1.74, 95% confidence interval (CI)=1.13-2.68). Persons with condition-specific functional loss secondary to UI were substantially more likely to have psychological distress as measured by the GHQ than were persons without UI (unadjusted OR=4.02, 95% CI=1.86 8.70). In multivariate models that controlled for potentially influential characteristics such as age, gender, ethnicity, and chronic medical conditions the association between condition-specific functional loss secondary to UI and psychological distress remained statistically significant. Among people with UI, persons with persistently elevated GHQ scores were much more likely to report condition-specific functional impairment from UI (adjusted OR=6.55, 95% CI=1.94 22.12). CONCLUSION: Individuals with UI, especially when incontinence was associated with condition-specific functional loss, were more likely to have psychological distress than were other older adults. Our findings support a general conceptual model that condition-specific functional impairment mediates the relationship between a chronic medical condition and psychological distress. PMID- 11943046 TI - Medical care inconsistent with patients' treatment goals: association with 1-year Medicare resource use and survival. AB - OBJECTIVES: To describe how frequently seriously ill persons perceive that the care they receive is inconsistent with treatment preferences and the effect on 1 year resource utilization. SETTING: Five U.S. teaching hospitals. DESIGN: Secondary analysis of interview data. PARTICIPANTS: Seriously ill Medicare beneficiaries. MEASUREMENTS: Interviews about patients' preferred approaches to care and whether they perceived care was consistent with these preferences. Part A and B costs for up to 1 year, adjusted for cost differences across hospitals and over time and for 1-year survival. RESULTS: Forty percent of the 1,185 study patients expressed a preference for treatment to focus on extending life, whereas 60% expressed a preference for comfort care. Eighty-six percent of the patients who wanted aggressive treatment reported that care was consistent with their preferences, but only 41% of those who preferred comfort care reported that care was consistent with their preferences. More than one-third of those with a preference for comfort care (35%) reported that the medical care that they received was inconsistent with their goals; 24% were unsure of treatment goals. Those who preferred comfort care but believed that their care was inconsistent with their wishes had higher estimated mean 1-year costs than those who believed that their care was consistent with their wishes (92,442 US dollars vs 52,098 US dollars, P < .001). Even after adjusting for differences in disease severity, age, gender, race, functional status, income, and years of education, adjusted costs were 1.4 times (95% confidence interval = 1.2-1.6) higher. However, 1-year survival was lower in these patients who stated that care was consistent with their preference to focus on comfort care than for those who wished to receive comfort care and stated that care was not consistent with their preference (38% vs 55% 1-year survival, P < .001). CONCLUSION: More than one in three seriously ill persons who prefer comfort care believe that their medical care is at odds with their preference that treatment focus on palliation. Such discord was associated with higher 1-year healthcare costs and increased survival. PMID- 11943047 TI - The last 48 hours of life in long-term care: a focused chart audit. AB - OBJECTIVES: As a component of palliative care educational program development, the faculty at the University of Ottawa Institute of Palliative Care wished to assess end-of-life care for patients in long-term care (LTC) settings to develop an educational strategy for physicians. DESIGN: A chart audit, focusing on the last 48 hours of life of residents dying in LTC facilities. SETTING: Five LTC facilities in a city in Canada. PARTICIPANTS: Residents who died in the LTC facilities in a 12-month period. Those who died suddenly (i.e., with no palliation period) or in a hospital were excluded. MEASUREMENTS: Symptoms highlighted in the literature as commonly found in the terminally ill and the matching treatments were recorded on an audit form created by the authors. Included were pain, dyspnea, noisy breathing, delirium, dysphagia, fever, and myoclonus. RESULTS: One hundred eighty-five charts were reviewed. A large number of patients were cognitively impaired. Cancer was the final diagnosis in 14% of cases. Respiratory symptoms were the most prevalent symptom, with dyspnea being first and noisy breathing third. Pain was second, with a prevalence similar to that found in studies of cancer patients. Dyspnea was not treated in 23% of the patients with this symptom; opioids were used in only 27% of cases with dyspnea. Ninety-nine percent of patients who experienced pain were treated for it. Less than one-third of patients with noisy breathing were treated. Delirium was not treated in 38% of the cases, and no anti-dopaminergic medications were administered. Nurses were primarily responsible for documenting end-of-life issues, supporting the families of the dying residents, and communicating with other team members. CONCLUSION: The focused chart audit identified the high prevalence of cognitive impairment in the patient population, which complicates symptom management. Respiratory symptoms predominated in the last 48 hours of life. This symptom profile differs from that of cancer patients, who, according to the literature, have more pain and less respiratory trouble. Management of symptoms was variable. Nurses played a crucial role in the care of dying residents through their documentation and communication of end-of-life issues. Appropriate palliative care education can provide knowledge and skills to all health-care professionals, including physicians, and assist them in the control of symptoms and improvement of quality of life for patients dying in LTC facilities. PMID- 11943048 TI - Does receipt of hospice care in nursing homes improve the management of pain at the end of life? AB - OBJECTIVES: To compare analgesic management of daily pain for dying nursing home residents enrolled and not enrolled in Medicare hospice. DESIGN: Retrospective, comparative cohort study. SETTING: Over 800 nursing homes in Kansas, Maine, Mississippi, New York, and South Dakota. PARTICIPANTS: A subset of residents with daily pain near the end of life taken from a matched cohort of hospice (2,644) and nonhospice (7,929) nursing home residents who had at least two resident assessments (Minimum Data Sets (MDSs)) completed, their last between 1992 and 1996, and who died before April 1997. The daily pain subset consisted of 709 hospice and 1,326 nonhospice residents. MEASUREMENTS: Detailed drug use data contained on the last MDS before death were used to examine analgesic management of daily pain. Guidelines from the American Medical Directors Association (AMDA) were used to identify analgesics not recommended for use in managing chronic pain in long-term care settings. The study outcome, regular treatment of daily pain, examined whether patients received any analgesic, other than those not recommended by AMDA, at least twice a day for each day of documented daily pain (i.e., 7 days before date of last MDS). RESULTS: Fifteen percent of hospice residents and 23% of nonhospice residents in daily pain received no analgesics (odds ratio (OR) = 0.57, 95% confidence interval (CI) = 0.45-0.74). A lower proportion of hospice residents (21%) than of nonhospice residents (29%) received analgesics not recommended by AMDA (OR = 0.65, 95% CI =0.52-0.80). Overall, acetaminophen (not in combination with other drugs) was used most frequently for nonhospice residents (25% of 1,673 prescriptions), whereas morphine derivatives were used most frequently for hospice residents (30% of 1,058 prescriptions). Fifty-one percent of hospice residents and 33% of nonhospice residents received regular treatment for daily pain. Controlling for clinical confounders, hospice residents were twice as likely as nonhospice residents to receive regular treatment for daily pain (adjusted odds ratio = 2.08, 95% CI = 1.68-2.56). CONCLUSION: Findings suggest that analgesic management of daily pain is better for nursing home residents enrolled in hospice than for those not enrolled in hospice.The prescribing practices portrayed by this study reveal that many dying nursing home residents in daily pain are receiving no analgesic treatment or are receiving analgesic treatment inconsistent with AMDA and other pain management guidelines. Improving the analgesic management of pain in nursing homes is essential if high-quality end-of-life care in nursing homes is to be achieved. PMID- 11943049 TI - Characteristics associated with fear of falling and activity restriction in community-living older persons. AB - OBJECTIVES: To identify the characteristics associated with restricting activity because of fear of falling (activity restriction) and to determine which characteristics distinguish older persons who restrict activity from those who have fear of falling but do not restrict their activities (fear of falling alone). DESIGN: Population-based cross-sectional study. SETTING: General community. PARTICIPANTS: One thousand sixty-four community-living persons aged 72 and older. MEASUREMENTS: Candidate predictors were identified from the following domains: demographic, health status, physical, psychosocial, and fall-related. The outcome measure was the report of no fear of falling, fear of falling alone,or activity restriction. RESULTS: Fifty-seven percent of the cohort reported no fear of falling, 24% reported fear of falling alone, and 19% reported restricting activity. The proportion of participants with poor health status, slow timed physical performance, activities of daily living disability, and poor psychosocial function was highest in those with activity restriction, intermediate in those with fear of falling alone, and lowest in those with no fear of falling. Of participants with fear of falling, characteristics independently associated with activity restriction were history of an injurious fall, slow timed physical performance, two or more chronic conditions, and depressive symptoms. CONCLUSION: Older persons who restrict activity are more physically frail and have a greater burden of chronic conditions and depressive symptoms than those who have fear of falling alone. These differences between persons with fear of falling may guide the refinement of clinical interventions and preventative programs. PMID- 11943050 TI - Prescription duration after drug copay changes in older people: methodological aspects. AB - OBJECTIVES: Impact assessment of drug benefits policies is a growing field of research that is increasingly relevant to health care planning for older people. Some cost-containment policies are thought to increase noncompliance. This paper examines mechanisms that can produce spurious reductions in drug utilization measures after drug policy changes when relying on pharmacy dispensing data. Reference pricing, a copayment for expensive medications above a fixed limit, for angiotensin-converting enzyme(ACE) inhibitors in older British Columbia residents, is used as a case example. DESIGN: Time series of 36 months of individual claims data. Longitudinal data analysis, adjusting for autoregressive data. SETTING: Pharmacare, the drug benefits program covering all patients aged 65 and older in the province of British Columbia, Canada. PARTICIPANTS: All noninstitutionalized Pharmacare beneficiaries aged 65 and older who used ACE inhibitors between 1995 and 1997 (N = 119,074). INTERVENTION: The introduction of reference drug pricing for ACE inhibitors for patients aged 65 and older. MEASUREMENTS: Timing and quantity of drug use from a claims database. RESULTS: We observed a transitional sharp decline of 110% t a standard error of 30% (P = .02) in the overall utilization rate of all ACE inhibitors after the policy implementation; five months later, utilization rates had increased, but remained under the predicted prepolicy trend. Coinciding with the sharp decrease, we observed a reduction in prescription duration by 31% in patients switching to no cost drugs. This reduction may be attributed to increased monitoring for intolerance or treatment failure in switchers, which in turn led to a spurious reduction in total drug utilization. We ruled out the extension of medication use over the prescribed duration through reduced daily doses (prescription stretching) by a quantity-adjusted analysis of prescription duration. CONCLUSION: The analysis of prescription duration after drug policy interventions may provide alternative explanations to apparent short-term reductions in drug utilization and adds important insights to time trend analyses of drug utilization data in the evaluation of drug benefit policy changes. PMID- 11943051 TI - Restraint use, restraint orders, and the risk of falls in hospitalized patients. AB - OBJECTIVES: To determine the relationship between physical restraints and falls in the acute hospital setting. DESIGN: Matched case-control study. SETTING: Inpatients at a 528-bed, urban, community based, acute care hospital. PARTICIPANTS: Two hundred twenty-eight patients who fell during hospitalization and 228 controls matched to cases by nursing unit and length of stay. MEASUREMENTS: Persons who fell were systematically evaluated at the time of fall by trained fall evaluators. For the cases, we sought to validate "orders for restraints" using "observed restraint use," defined as the use of restraints at the time of fall as determined through direct observation or interviews with nursing staff. RESULTS: Patients with orders for restraints were more likely to fall than patients without orders for restraints (multivariate relative risk = 6.3, 95% confidence interval (CI) = 1.8-22.3). However, in the cases, there was poor correlation between "orders for restraints" and "observed restraint use" at the time of fall (kappa = 0.15, 95% CI-0.4-0.34). CONCLUSION: Because orders for restraint use may not reflect actual restraint use at the time of a fall, observational studies relating use of restraints to the risk of falls should be interpreted with caution. Despite this caveat, we could find no evidence that restraints protect hospitalize patients from falling. PMID- 11943052 TI - The GPCOG: a new screening test for dementia designed for general practice. AB - OBJECTIVES: To design and test a brief, efficient dementia-screening instrument for use by general practitioners (GPs). DESIGN: The General Practitioner Assessment of Cognition (GPCOG) consists of cognitive test items and historical questions asked of an informant. The validity of the measure was assessed by comparison with the criterion standard of diagnoses of dementia derived from the Diagnostic and Statistical Manual of Mental Disorders (Fourth Edition). SETTING: Primary care doctors' offices. PARTICIPANTS: Sixty-seven GPs administered the GPCOG to 283 community-dwelling patients aged 50 to 74 with memory complaints or aged 75 and older. MEASUREMENTS: The Cambridge Mental Disorder of the Elderly Examination, the Abbreviated Mental Test (AMT), the Mini-Mental State Examination (MMSE), the 15-item Geriatric Depression Scale, and the 12-item Short-Form Health Survey. RESULTS: The GPCOG was reliable and superior to the AMT (and possibly to the MMSE) in detecting dementia. The two-stage method of administering the GPCOG (cognitive testing followed by informant questions if necessary)had a sensitivity of 0.85, a specificity of 0.86, a misclassification rate of 14%, and positive predictive value of 71.4%. Patient interviews took less than 4 minutes to administer and informant interviews less than 2 minutes. The instrument was reported by GPs to be practical to administer and was acceptable to patients. CONCLUSION: The GPCOG is a valid, efficient, well-accepted instrument for dementia screening in primary care. PMID- 11943053 TI - Xerostomia and the geriatric patient. AB - Saliva is essential for the preservation of oral-pharyngeal health, and disorders of salivary physiology are associated with numerous oral and pharyngeal problems, particularly in older people. Although salivary function is remarkably intact in healthy aging, medical problems, medications, and head and neck radiotherapy can cause salivary dysfunction and complaints of xerostomia among older people. Sjogren's syndrome, an autoimmune exocrinopathy, is the most common medical disease associated with salivary dysfunction. Medications with anticholinergic side effects will impair salivary output, and head and neck radiotherapy for cancer will cause permanent destruction of salivary glands. Treatments for salivary problems are based upon establishing a diagnosis, protecting oral and pharyngeal health, stimulating remaining glands, and replacing lost salivary fluids. PMID- 11943054 TI - Refusing artificial nutrition and hydration: does statutory law send the wrong message? AB - Ethical consensus and appellate court decisions view artificial nutrition and hydration (ANH) as medical treatment that can be refused like other treatments. However, advance directive statutes may produce obstacles for refusal of ANH, as distinct from other life-sustaining treatments, in patients who lack capacity. This paper reviews state statutes and appellate case law regarding medical decision making for patients who lack decisional capacity. Twenty states (39%) have one or more explicit statutory provisions delineating a separate and more stringent standard for ANH refusal. These standards include higher evidentiary standard; requirement for specific preauthorization, qualifying medical conditions, second medical opinion, or judicial review; refusal not permitted; refusal not permitted if death would result from "starvation" or "dehydration"; and previous law with higher standard applies to old documents. In 11 of these states and in eight others, statutory law contains language that could be misinterpreted, implying, but not rising to, an explicitly higher standard. Four appellate decisions departed from the judicial consensus that ANH can be refused like other treatments, but subsequent court decisions or legislative enactments reduced or eliminated their impact. Legislators and the courts should ask whether higher standards for ANH refusal are appropriate in light of case law authority that ANH should not be treated differently and in light of statutory language that preserves those common law rights. These higher standards may make it more difficult in certain states to refuse ANH for patients who lack capacity or place a burden on good practice by making providers fearful of the law. PMID- 11943055 TI - Care of patients nearing death: another view. AB - A person with an advanced, progressive, ultimately fatal illness faces multifaceted suffering. It is difficult to be sick. Uncontrollable deterioration of one's body can be horrible. Many people detest seeking help from others, yet nearly all want companionship in the face of death. The choice is hard, between certain death, even if it is promised to be comfortable and dignified, and hope, even if it is a burdensome treatment that offers small chance of limited benefit. Dying patients are often expected to make such painful choices. Perhaps what is most painful is to be dying. A person whose near future necessarily contains all this will often need a trusted, unhurried companion, acting with gentle advocacy and humility. This paper offers perspective and suggestions to such companions/advocates. PMID- 11943056 TI - Sensitivity and specificity of cognitive and functional screening instruments for dementia: the Indo-U.S. Dementia Epidemiology Study. AB - There is a shortage of adequate screening instruments for dementia in poorly educated populations and non-English-speaking groups. An epidemiological survey was conducted in a population-based, largely illiterate, sample of 5,126 individuals aged 55 and older in 28 villages in the rural community of Ballabgarh in northern India. All participants were administered a general mental status test, the Hindi Mental State Examination (HMSE), and a brief battery of neuropsychological tests. Their informants answered a questionnaire assessing functional ability, the Everyday Abilities Scale for India (EASI). Six hundred thirty-two participants underwent clinical diagnostic evaluation for dementia. We investigated whether the sensitivity, specificity, and predictive value for dementia of the mental status test could be improved by the addition of the brief neuropsychological test battery or the functional questionnaire, comparing the instruments alone and in combination. In participants who could be tested cognitively, the HMSE, the neuropsychological battery, and EASI had sensitivities of 81.3%, 81.3%, and 62.5%, respectively, with specificities of 60.2%, 74.5%, and 89.7%, respectively. The combination of all three was 93.8% sensitive and 41.8% specific. The sensitivity of the HMSE alone was nonsignificantly improved by the addition of either the EASI or the neuropsychological battery, whereas its specificity was significantly decreased by either addition. An advantage of the EASI was that it could also be administered to informants of subjects who were cognitively untestable. In this largely illiterate community, with a low prevalence of dementia, the combination of cognitive tests and a functional ability questionnaire had substantial value for population screening. PMID- 11943057 TI - A comparison of two methods for identifying frail Medicare-aged persons. AB - This article compares the efficacy of two screening methods to identify frail Medicare-aged persons using self-report questionnaire data: a clinical judgment method developed by nurse and social worker professionals in a community-based long-term care department and an empirical research method previously developed by the Center for Health Research using computer formulas and stepwise logistic regression coefficients. A sub-aim was to see whether the empirical method proved robust over time by measuring aggregate utilization and mortality in frail and nonfrail cohorts, which would increase the interest of physicians, managed care organizations, and other agencies providing service to Medicare beneficiaries. PMID- 11943058 TI - Infections in residents of long-term care facilities: an agenda for research. Report of an expert panel. AB - The number of frail, older residents of long-term care facilities (LTCFs) will increase dramatically over the next 30 years. Improving the quality of health care provided in LTCFs is an important national and international priority. Improving the prevention and management of infections in LTCFs is a critical component of efforts to improve quality of care and poses unique challenges. This report summarizes the presentations and discussions of participants in an invitational conference to propose a research agenda for prevention and management of infections in LTCFs. The conference was held in March 2001 in Atlanta, Georgia. The discussants identified key research questions to better understand general issues involving the overall burden of infections in LTCFs, prevention and control interventions, and antimicrobial use and resistance. The participants also discussed research questions involving specific infections, including pneumonia and urinary tract, skin, and soft tissue infections. Recommendations for research were discussed and are presented in summary form in this report. Improving the prevention and management of infections in LTCF residents should be a priority if quality of care in these facilities is to be improved. Many unanswered questions remain in this field, and the research agenda outlined in this report will require resources and focus. The benefit of such efforts to LTCF residents and their caregivers is likely to be substantial. PMID- 11943059 TI - An evidence-based approach to the diagnosis of deep vein thrombosis: beyond the ultrasound report. PMID- 11943060 TI - Management of diabetes mellitus in older adults: are national guidelines appropriate? PMID- 11943061 TI - Oral health is cost-effective to maintain but costly to ignore. PMID- 11943062 TI - Regulating the foregoing of artificial nutrition and hydration: first, do some harm. PMID- 11943063 TI - Reducing delirium after hip fracture. PMID- 11943064 TI - Aspiration pneumonia and altered dental status. PMID- 11943065 TI - Are the unsafe combinations always inappropriate? PMID- 11943066 TI - Outpatient geriatric evaluation and management. PMID- 11943067 TI - Factors regulating Hb F synthesis in thalassemic diseases. AB - BACKGROUND: The thalassemic syndromes originate from mutations of the globin genes that cause, besides the characteristic clinical picture, also an increased Hb F amount. It is not yet clear if there are more factors, besides the beta globin genotype, determining the Hb F production. We have tried to find out if there are relations between total Hb and Hb F, between erythropoietin (Epo) and Hb F, between Hb F and point mutations of the gamma gene promoters. MATERIALS AND METHODS: Hematologic parameters, iron status, alpha/non-alpha globin ratio, Epo level, and thalassemic defects of the alpha-, beta-, and gamma-globin genes were explored using standard methods in patients affected by thalassemic diseases. Ninety-five non thalassemic individuals have been examined as controls. RESULTS: Two clinical variants of beta-thalassemia intermedia referred to as beta-thal int sub-silent and evident are associated with distinct sets of mutations of the beta globin gene. Silent beta thal mutations are invariably associated with sub-silent beta thal int; beta degrees or severe beta+ thal mutations are associated with evident beta thal int (88%) and almost invariably (98%) with thalassemia major. A positive correlation was observed between the severity of the disease and the Hb F level, but no correlation was found between the Hb F and erythropoietin (Epo) level. The mutation Ggamma -158 C→T was detected in 26.9% of patients affected by beta-thal int sub-silent and evident, respectively, but only in 2% of patients with thalassemia major. CONCLUSIONS: The severity of beta-thal int and the increased Hb F level are strictly dependent from the type of beta-globin gene mutations. No relation is found between Hb F synthesis and Epo secretion. The mutation Ggamma -158 C→T, common among patients affected by beta-thal int and very rare in thal major patients, does not seem, in this study, to influence the Hb F content in beta thal int patients. PMID- 11943068 TI - Immunity status against poliomyelitis in Germany: determination of cut-off values in International Units. AB - BACKGROUND: To prevent importations of wild polioviruses into a polio free region a high level of population immunity must be kept. Standard methodology for determination of polio antibodies is a feature aimed at obtaining consistent results. An International Standard Serum for polio antibodies exists, but no protective level in International Units is defined. METHODS: A representative study was carried out in order to determine the serological status against poliomyelitis in Germany (n = 2564, age 18-79 years). Furthermore, sera from persons aged less than 18 years were included (n = 881). Microneutralization test has been used for determination of antibody levels. Results have been expressed in International Units. RESULTS: The results of this study indicate that the cut off level for polio antibodies is 0.075 IU/ml for Polio 1, 0.180 IU/ml for Polio 2 and 0.080 IU/ml for Polio 3. Neutralizing antibodies against poliovirus type 1, 2 and 3 were detected in 96.2%, 96.8% and 89.6% of samples, respectively. CONCLUSIONS: Overall, this seroprevalence indicates a very high level of immunity of the general population. It must be kept after the switch of immunization strategy from attenuated to inactivated vaccine in Germany. PMID- 11943069 TI - The Caenorhabditis elegans Y87G2A.14 Nudix hydrolase is a peroxisomal coenzyme A diphosphatase. AB - BACKGROUND: The number of Nudix hydrolase family members varies widely among different organisms. In order to understand the reasons for the particular spectrum possessed by a given organism, the substrate specificity and function of different family members must be established. RESULTS: The Y87G2A.14 Nudix hydrolase gene product of Caenorhabditis elegans has been expressed as a thioredoxin fusion protein in Escherichia coli and shown to be a CoA diphosphatase with catalytic activity towards CoA and its derivatives. The products of CoA hydrolysis were 3',5'-ADP and 4'-phosphopantetheine with Km and kcat values of 220 microM and 13.8 s(-1) respectively. CoA esters yielded 3',5' ADP and the corresponding acyl-phosphopantetheine. Activity was optimal at pH 9.5 with 5 mM Mg2+ and fluoride was inhibitory with a Ki of 3 microM. The Y87G2A.14 gene product has a potential C-terminal tripeptide PTS1 peroxisomal targeting signal - SKI. By fusing a Y87G2A.14 cDNA to the C-terminus of yeast-enhanced green fluorescent protein, the enzyme appeared to be targeted to peroxisomes by the SKI signal when transfected into yeast cells. Deletion of SKI abolished specific targeting. CONCLUSIONS: The presence of related sequences with potential PTS1 or PTS2 peroxisomal targeting signals in other organisms suggests a conserved peroxisomal function for the CoA diphosphatase members of this group of Nudix hydrolases. PMID- 11943070 TI - Detection of fast light-activated H+ release and M intermediate formation from proteorhodopsin. AB - BACKGROUND: Proteorhodopsin (pR) is a light-activated proton pump homologous to bacteriorhodopsin and recently discovered in oceanic gamma-proteobacteria. One perplexing difference between these two proteins is the absence in pR of homologues of bR residues Glu-194 and Glu-204. These two residues, along with Arg 82, have been implicated in light-activated fast H+ release to the extracellular medium in bR. It is therefore uncertain that pR carries out its physiological activity using a mechanism that is completely homologous to that of bR. RESULTS: A pR purification procedure is described that utilizes Phenylsepharose and hydroxylapatite columns and yields 85% (w/w) purity. Through SDS-PAGE of the pure protein, the molecular weight of E.-coli-produced pR was determined to be 36,000, approximately 9,000 more than the 27,000 predicted by the DNA sequence. Post translational modification of one or more of the cysteine residues accounts for 5 kDa of the weight difference as measured on a cys-less pR mutant. At pH 9.5 and in the presence of octylglucoside and diheptanoylphosphotidylcholine, flash photolysis results in fast H+ release and a 400-nm absorbing (M-like) photoproduct. Both of these occur with a similar rise time (4-10 micros) as reported for monomeric bR in detergent. CONCLUSIONS: The presence of fast H+ release in pR indicates that either different groups are responsible for fast H+ release in pR and bR (i.e. that the H+ release group is not highly conserved); or, that the H+ release group is conserved and is therefore likely Arg-94 itself in pR (and Arg-82 in bR, correspondingly). PMID- 11943071 TI - Potential antimutagenic activity of berberine, a constituent of Mahonia aquifolium. AB - BACKGROUND: As part of a study aimed at developing new pharmaceutical products from natural resources, the purpose of this research was twofold: (1) to fractionate crude extracts from the bark of Mahonia aquifolium and (2) to evaluate the strength of the antimutagenic activity of the separate components against one of the common direct-acting chemical mutagens. METHODS: The antimutagenic potency was evaluated against acridine orange (AO) by using Euglena gracilis as an eukaryotic test model, based on the ability of the test compound/fraction to prevent the mutagen-induced damage of chloroplast DNA. RESULTS: It was found that the antimutagenicity of the crude Mahonia extract resides in both bis-benzylisoquinoline (BBI) and protoberberine alkaloid fractions but only the protoberberine derivatives, jatrorrhizine and berberine, showed significant concentration-dependent inhibitory effect against the AO induced chloroplast mutagenesis of E. gracilis. Especially berberine elicited, at a very low dose, remarkable suppression of the AO-induced mutagenicity, its antimutagenic potency being almost three orders of magnitude higher when compared to its close analogue, jatrorrhizine. Possible mechanisms of the antimutagenic action are discussed in terms of recent literature data. While the potent antimutagenic activity of the protoberberines most likely results from the inhibition of DNA topoisomerase I, the actual mechanism(s) for the BBI alkaloids is hard to be identified. CONCLUSIONS: Taken together, the results indicate that berberine possesses promising antimutagenic/anticarcinogenic potential that is worth to be investigated further. PMID- 11943072 TI - Effect of a homeopathic drug, Chelidonium, in amelioration of p-DAB induced hepatocarcinogenesis in mice. AB - BACKGROUND: Crude extracts of Chelidonium majus, and also purified compounds derived from crude extracts of this plant, have been reported to exhibit anti viral, anti-inflammatory, anti-tumor and anti-microbial properties both in vitro and in vivo. Chelidonium is a homeopathic drug routinely used against various liver disorders including cancer in humans. Two potencies of Chelidonium (Ch-30, Ch-200) have been tested for their possible anti-tumor and enzyme modulating activities in liver and anti-clastogenic effects during p-DAB-induced hepatocarcinogenesis in mice compared to suitable controls. METHODS: Several cytogenetic and enzymatic protocols were used at three fixation intervals; at 60 days, 90 days and 120 days of treatment. Different sets of healthy mice were fed: i) hepatocarcinogen, p-DAB plus phenobarbital (PB), ii) only PB, iii) neither p DAB nor PB (normal control). One set of mice fed with p-DAB plus PB was also fed Ch-30 (iv) and another set Ch-200 (v). All standard currently used methods were adopted for cytogenetical preparations and for the enzyme assays. RESULTS: All group (i) mice developed tumors in liver at all fixation intervals, while none of group (ii) and (iii) mice developed any tumors. About 40% mice in group (iv) and group (v) did not show tumor nodules in their liver. Feeding of Chelidonium to group (iv) and (v) mice reduced genotoxic effects to a significant extent (p < 0.05 to p < 0.001). CONCLUSION: The homeopathic drug Chelidonium exhibited anti tumor and anti-genotoxic activities and also favorably modulated activities of some marker enzymes. Microdoses of Chelidonium may be effectively used in combating liver cancer. PMID- 11943073 TI - Diclofenac does not interact with codeine metabolism in vivo: a study in healthy volunteers. AB - BACKGROUND: Previously, we have demonstrated a marked inhibition of codeine glucuronidation by diclofenac in human liver tissue homogenate. We therefore aimed to investigate whether diclofenac inhibits glucuronidation of codeine also in vivo in healthy volunteers. METHODS: In a randomised, placebo-controlled, double-blind, cross-over study, 12 healthy volunteers received a singe of 100 mg codeine phosphate plus 50 mg diclofenac sodium or codeine phosphate plus placebo. Over a 36 hour period serum concentrations of codeine and its metabolites as well as urinary excretion were analysed using LC-mass spectrometry. Side effects were recorded and analgesic efficacy was determined using the cold pressor test (0-6 h). RESULTS: A single dose of diclofenac did not alter the formation of codeine-6 glucuronide in healthy volunteers. Metabolic clearance of codeine to morphine was not affected by diclofenac. In terms of side effects, both treatments were well tolerated. Diclofenac did not significantly influence the analgesic effects of codeine in the cold pressor test. CONCLUSIONS: In contrast to recent in vitro data, a single oral dose of diclofenac did not alter the glucuronidation of codeine in healthy volunteers. PMID- 11943074 TI - Endocrine stress response in jugular-vein cannulated rats upon multiple exposure to either diethyl-ether, halothane/O2/N2O or sham anaesthesia. AB - The main objective of this study was to assess the endocrine stress response to multiple anaesthesia followed by sham anaesthesia in order to detect any memory effects. For this purpose, jugular-vein cannulated rats were subjected to either sham, diethyl-ether or halothane/O2/N2O anaesthesia, and their plasma ACTH, corticosterone, glucose, adrenaline and noradrenaline levels measured. The study had three separate experiments, each consisting of a control and treatment group. In two experiments, the rats were exposed to high or low concentrations (40-15%) of diethyl ether, using either a jar containing cotton soaked in diethyl ether or a vaporizer. In the third experiment, rats were exposed to halothane/O2/N2O. Control animals underwent sham anaesthesia. Blood samples were taken 6 min before and at 5, 15 and 55 min after starting the exposure (t = 0 min). For each variable, the dt5 (level at t = 5 min minus that at t = -6 min) and the cumulative levels over the one-hour period as determined by the area under the curve (AUC) were calculated. Further, the peak levels (Cmax) were determined. The mean time needed to induce anaesthesia was 68, 121 and 55 s for exposure to high and low concentrations of diethyl ether and to halothane/O2/N2O, respectively. Increased noradrenaline and adrenaline dt5 levels were observed only after the first exposure to the high concentration of diethyl ether. Multiple anaesthesia sessions using either diethyl ether or halothane/O2/N2O did not clearly influence adrenaline and noradrenaline levels. Diethyl ether induced a sharp rise in plasma ACTH and glucose levels, irrespective of the concentration used. The response of the ACTH and glucose was similar for single and multiple exposure. An increased response of ACTH, corticosterone and glucose to sham anaesthesia following multiple induction of anaesthesia was observed for the high concentration of diethyl ether only. Halothane/O2/N2O raised plasma glucose without differences between single and multiple anaesthesia sessions. Upon sham anaesthesia following multiple exposures to halothane/O2/N2O, glucose levels were significantly increased. This study indicates that repeated anaesthesia in rats can elicit an increased stress response during subsequent handling and change of environment. PMID- 11943075 TI - Effects of CO2 anaesthesia on central nervous system activity in swine. AB - The objective of the study was to examine the changes in central nervous system (CNS) activity and physical behaviour during induction and awakening from CO2 anaesthesia. Two studies, each using pigs immersed into 90% CO2 gas for a period of 60 s were performed. In study 1, we monitored middle latency auditory evoked potentials (changes in latencies, amplitudes and a depth of anaesthesia index), electroencephalographic parameters (delta, theta, alpha and beta electroencephalographic power and 95% spectral edge frequency) and heart rate; and in study 2, we monitored body movements and arterial and venous partial pressure of CO2 and O2. No behavioural signs of distress were observed during the early part of the induction. The swine exhibited muscular activity from 13-30 s after induction-start as well as during awakening from anaesthesia, possibly because of a transitory weaker suppression of the brain stem than of the cortex. The CNS and blood gas parameters started to change from the very start of induction. The CNS suppression lasted only approximately one minute after the end of the induction period. The two studies indicated a good temporal relationship between changes in amplitude, depth of anaesthesia index, spectral edge frequency, and arterial PCO2 during the induction period. PMID- 11943076 TI - Regional distribution of glutamate in the central nervous system of rat terminated by carbon dioxide euthanasia. AB - Carbon dioxide euthanasia is an established method for the termination of small laboratory animals. It has also been employed by the authors in neurobiological research on the post-mortem glutamate concentration in the structures of rat brains. The following investigations were aimed at optimizing the termination procedure based on the CO2 saturation rate of the inhaled air. Two rates of CO2 flow were applied, and the higher one significantly augmented the glutamate level in the hippocampus and cerebellum. The relationship between this finding and signs of central fear reaction is discussed. The authors conclude that lower rather than higher CO2 flow in euthanasia procedures is gentler and is therefore preferable for use with laboratory animals. PMID- 11943077 TI - Oral buprenorphine and aspirin analgesia in rats undergoing liver transplantation. AB - The objective of this study was to establish effective postoperative analgesia for Dark Agouti rats undergoing liver transplantation with minimal additional stress due to handling and no adverse effect on transplant outcome. Oral administration of buprenorphine (0.5 mg/kg/dose) or aspirin (100 mg/kg/dose) in raspberry-flavoured gelatine were compared to controls receiving no treatment or plain gelatine. The drugs were presented five times: immediately on recovery from anaesthesia and at 12 h intervals thereafter. All rats underwent right nephrectomy and replacement of their liver by an arterialized liver isograft preserved optimally for 24 h. All groups had reversible hepatic damage, lost weight and demonstrated severely reduced dark cycle activity after surgery. Neither treatment appeared to ameliorate the loss of body weight that probably reflected hepatic insufficiency during the first week as well as pain and surgical stress. In the second week, when liver function was 'normal', rats began to regain weight at the pre-transplant rate. Aspirin treatment significantly increased activity during the first and second dark cycles after surgery, whereas buprenorphine significantly increased activity during the second dark cycle only. Neither drug had any apparent adverse effects on the rats or on graft function. Postoperative oral administration of aspirin should be incorporated into future programmes of liver transplantation in rodents. More effective treatment in the immediate postoperative period may require oral administration of analgesia prior to surgery or a single subcutaneous injection of an analgesic agent on completion of surgery in addition to postoperative oral administration of aspirin. PMID- 11943078 TI - Assessment of the postoperative discomfort of intra-auricularly hypophysectomized rats. AB - Rats subjected to hypophysectomy make up one of the largest groups of experimental animals in Europe, since there is a legal demand for batch testing of industrially produced growth hormones. To describe the clinical performance of rats having undergone hypophysectomy, animals were examined postoperatively by monitoring behaviour, body temperature and food intake. Behavioural changes were observed in rats that had only been anaesthetized, as well as in sham-operated rats, while no behavioural deviations could be shown in hypophysectomized rats. On the first day after surgery all rats had declining body temperature and food intake; and this change was not reversed by treatment with carprofen, buprenorphine or oxytetracycline. The mortality rate in rats treated with buprenorphine was increased, as was the mortality rate in rats hypophysectomized when weighing more than 100 g. As there seemed to be no differences whether methohexital or a combination of fentanyl, fluanison and midazolam was used, the latter anaesthesia is recommended due to its analgesic potential. For post surgical analgesic treatment, carprofen is recommended rather than buprenorphine. At best, the use of hypophysectomized rats should be replaced in industrial batch testing by an existing in vitro method. PMID- 11943079 TI - Serum alpha2-macroglobulin and cytokine measurements in an acute inflammation model in rats. AB - An enzyme-linked immunosorbent assay (ELISA) for rat alpha2-macroglobulin (alpha2M) using a monoclonal antibody was developed, and used to measure alpha2M in sera from rats injected intramuscularly with turpentine oil as an inflammatory agent. The mean concentration of alpha2M gradually increased and peaked 2 days after the turpentine oil injection. The peak alpha2M concentration ranged from 2362-8472 microg/ml (mean 4531 microg/ml), which was 50-290 times higher than the pre-dosing levels of 23-61 microg/ml. In addition, interleukins (IL)-1, IL-2, IL 4, IL-6, IL-8, IL-10, and interferon (IFN)-gamma were measured using commercial ELISA reagent kits. IL-6 and IL-8 increased and peaked 12 h after turpentine oil injection, the levels being 5-51 times and 2-38 times the pre-dosing ones, respectively. The concentrations of IL-1, IL-2, IL-4, IL-10 and IFN-gamma in rats injected with turpentine oil did not change. PMID- 11943080 TI - Failure of liver cirrhosis induction by thioacetamide in Nagase analbuminaemic rats. AB - Repeated administration of thioacetamide (TA), either intraperitoneally or in drinking water, produced liver cirrhosis in normal Sprague-Dawley rats (SDR) with significant histological alterations similar to those observed in human cirrhosis. In the present study, we evaluated the ability of TA to induce liver cirrhosis in mutant Nagase analbuminaemic SDR. Thioacetamide was administered either intraperitoneally up to 4 months or in drinking water up to 6 months to normal and to Nagase analbuminaemic SDR. Nagase analbuminaemic rats (NAR) were also administered TA in drinking water up to 10 months. Liver cirrhosis development was determined by macroscopic and microscopic analysis. In contrast to normal SDR, no histological characteristics of cirrhosis could be observed in NAR submitted to a 4 or 6 months treatment with TA. Such failure to induce cirrhosis in Nagase rats was confirmed even after prolonged TA administration in drinking water for up to 10 months. In contrast, fibrosis and cholangiolar proliferation occurred in the 10-month TA-treated analbuminaemic rats, suggesting that the mechanisms involved in cirrhosis induction are different from those involved in fibrosis development and carcinogenesis. It is unlikely that the protective effect against TA-induced cirrhosis observed in analbuminaemic rats is related to the absence of albumin in this rat strain, since a co-administration of TA with albumin in analbuminaemic rats did not restore the potential for TA to induce cirrhosis in this rat strain. In conclusion, the fact that induction of cirrhosis by TA is prevented in the inherently hyperlipidaemic and hypercholesterolaemic analbuminaemic rats could be considered for potential application in the treatment of clinical cirrhosis. PMID- 11943081 TI - Validation of transrectal ultrasonographic volumetry for orthotopic prostate tumours in mice. AB - Orthotopic human prostate tumour models in athymic nude mice are regarded as being most suitable for fundamental and pre-clinical research on prostate cancer. The anatomic localization of the tumour in the pelvis, however, provides little possibility for monitoring tumour growth or regression. To assess time-related changes in orthotopic tumour volume, we applied transrectal ultrasonography (TRUS) to the murine prostate. This technique has the advantages of allowing accurate monitoring of tumours during therapeutic manipulations and a reduction of animal use due to a reduction of sacrificing endpoints. To validate the TRUS method, the mouse prostate reconstitution model, RM-9, and the prostate-specific antigen (PSA) producing human prostate cancer xenograft PC-346 were used. Volumetric calliper measurements were performed with a 30 MHz ultrasound probe designed for intra-arterial use in humans. Tumour weight, determined at various time-points, was found to be closely related to actual tumour weight (R = 0.99) and, in the PC-346 model, to the level of PSA in the plasma. Furthermore, the interobserver variation for TRUS was low for tumours above 50 mg. Thus, TRUS for murine prostate tumours proves to be an accurate, reproducible and sensitive method. PMID- 11943082 TI - A new animal model of femoral head necrosis induced by intraosseous injection of ethanol. AB - There is no reliable animal model of the early stages of osteonecrosis of the femoral head (ONFH) for the evaluation of new therapeutic approaches. In this study, we propose a new animal model of femoral head osteonecrosis. Pure ethanol was injected into the centre of the femoral head in adult Merino sheep under fluoroscopic control. After 3, 6 and 12 weeks the animals were killed and the femoral heads were harvested. Microradiographic and histological changes were analysed and recorded. Partial necrosis was documented over a period of 12 weeks in all animals. The appearance of necrosis in combination with intact macrotexture, macrocirculation and joint cartilage is similar to the features described in early ONFH in humans. Due to its efficacy and its similarity to the early stages of ONFH in humans, this model may be suitable to evaluate new therapeutic techniques in the treatment of ONFH. PMID- 11943083 TI - Intravenous tolerability of an acid vehicle in Sprague Dawley rats and beagle dogs after daily slow bolus injection and infusion for one hour during 14 days. AB - To assess the tolerability of an acid vehicle to be used in toxicology studies, a low pH aqueous solution containing 16.4 mg/ml of citric acid, 4.2 mg/ml of disodium phosphate, 25 mg/ml of mannitol, adjusted with phosphoric acid/NaOH 1 M to pH 3 was daily administered intravenously to rats and dogs for 14 consecutive days. The dosing regimen consisted of a slow intravenous bolus injection given over 30 s (0.75 and 0.625 ml/kg, for rats and dogs, respectively) followed by intravenous infusion for one hour (3.75 and 2.75 ml/kg/h, for rats and dogs, respectively). In rats, the dose was administered via the lateral tail vein. In dogs, the intravenous bolus dose was administered via the vena cephalica, vena saphena or vena jugularis, whilst the infusion dose was given into the vena cephalica or vena saphena. In rats, administration of the vehicle was associated with clinical signs (occasional mild vocalization and agitation) which were considered to be due to local irritation during the dosing procedure. Nevertheless, only mild histopathological changes at the injection site were found, while no relevant clinical chemistry changes were found in this species. However, the vehicle caused significant vascular damage with thrombus formation in the dog. It is therefore concluded that this vehicle is suitable for 2-week rat toxicity studies, if carefully applied. The vehicle with its present regimen should not be used in dogs, in view of the prohibitive findings. PMID- 11943084 TI - The effect of daily disturbance on the breeding performance of mice. AB - The United Kingdom Home Office Code of Practice for the housing and care of breeding animals requires that, 'the general well-being of all animals must be checked at least once daily'. However, excessive daily disturbance of rodent breeding colonies could be counter-productive to animal welfare if it increases pre-weaning mortality. An experiment involving 100 breeding cages of BALB/c mice compared daily inspection of the mouse cages, but without disturbing the mice within the nest, with daily inspection in which every individual was studied even if this involved removing the cage lid and disturbing the nest. No statistically significant differences were found between the two groups in breeding performance or pre-weaning mortality, though the disturbed group produced marginally fewer offspring and had slightly higher mortality. Average weaning weight did not differ between the groups, but sexual dimorphism at weaning was significantly increased in the disturbed group. It is concluded that there are unlikely to be any welfare benefits in an inspection regimen that involves disturbance of breeding mice, provided the cage is inspected daily. PMID- 11943085 TI - Fluctuating asymmetry in mice and rats: evaluation of the method. AB - Fluctuating asymmetry, which reflects small, random deviations from symmetry in otherwise bilaterally symmetrical characters, may be used as an indicator of developmental instability in humans and farm animals, and it may also be applicable as a stress indicator. We intended to find a method to allow the use of fluctuating asymmetry as a stress indicator in laboratory animals. That method had to be reproducible and reliable. Furthermore, its applicability in laboratory animals would be improved if it was possible to obtain measurements on the skin surface that correlated with results obtained by measuring the skeleton directly. Seven traits in mice and five traits in rats were evaluated for their applicability for measuring fluctuating asymmetry in mice and rats. Two out of the seven traits, i.e. the width of the joint between the third metatarsal bone and the digital bone on the hind paw, and the length of the incisor tooth at the top, were found to be reliable and reproducible for detecting fluctuating asymmetry in mice as well as in rats. Three out of the seven traits, i.e. the width of the carpal bones, the width of the joint between the tibia and the tarsal bones, and the length of the incisor tooth at the bottom, did express fluctuating asymmetry, but showed a poor day-to-day reproducibility. If the day to-day reproducibility could be increased, these three traits might also be suitable for measuring fluctuating asymmetry in mice and rats. The last two traits, i.e. the length of ulna and the length of calcaneus plus metatarsal bone i.v., measured both on the skin surface and directly on the bone, did not express fluctuating asymmetry, and had a poor day-to-day reproducibility. These two traits are not suitable for measuring fluctuating asymmetry in mice and rats. PMID- 11943086 TI - PCR for the detection of Streptobacillus moniliformis. AB - Streptobacillus moniliformis is a Gram-negative bacterium found in various laboratory animal species and is the cause of rat bite fever and Haverhill fever in man. In order to evaluate a polymerase chain reaction (PCR) for the detection of this zoonotic bacterium in animal tissues a set of primers was designed based on the DNA base sequence of part of the 16S rRNA gene from 11 S. moniliformis strains. The PCR detected as few as 2-6 copies of S. moniliformis DNA. A 296 bp DNA fragment was amplified from S. moniliformis strains from rodents, humans and turkeys. Amplicons of about the same size were obtained from Fusobacterium necrogenes and Sebaldella (Bacteroides) termitidis but Bfa I treatment of these amplicons did not result in the S. moniliformis specific 130 bp DNA fragment. The in silico evaluation of 14 additional Fusobacterium spp. and 12 unculturable phytoplasmas indicated that none is likely to give rise to confusing amplicons or DNA fragments. The PCR detected S. moniliformis infection in all four orally- and four intravenously-infected C57BL/6 mice and the bacterium was cultured from all but one mouse. The PCR detected S. moniliformis infection in all 12 orally infected WU rats, and in five of eight rats exposed to natural infection. Enzyme linked immunosorbent assay (ELISA) and PCR were equally successful in detecting infection in rats but S. moniliformis was not detected by using culture. PMID- 11943087 TI - Carbon dioxide concentrations in unventilated IVC cages. AB - The use of individually ventilated cage (IVC) systems has become more common worldwide. The various systems are becoming more and more sealed in order to protect the animals against infections and the staff against allergens; which, however, may lead to problematic CO2 concentrations, if the cages are left unventilated. In this study it is shown that, depending on how tight the cage is and the number of animals housed in each cage, CO2 inside the cage within 2 h will increase to levels of between 2 and 8%. PMID- 11943089 TI - Combined data, Bayesian phylogenetics, and the origin of the New Zealand cicada genera. AB - We have applied Bayesian and maximum likelihood methods of phylogenetic estimation to data from four mitochondrial genes (COI, COII, 12S, and 16S) and a single nuclear gene (EF1alpha) from several genera of New Zealand, Australian, and New Caledonian cicada taxa. We specifically focused on the heterogeneity of phylogenetic signal among the different data partitions and the biogeographic origins of the New Zealand cicada fauna. The Bayesian analyses circumvent many of the problems associated with other statistical tests for comparing data partitions. We took an information-theoretic approach to model selection based on the Akaike Information Criterion (AIC). This approach indicated that there was considerable uncertainty in identifying the best-fit model for some of the partitions. Additionally, a large amount of uncertainty was associated with many parameter estimates from the substitution model. However, a sensitivity analysis on the combined dataset indicated that the model selection uncertainty had little effect on estimates of topology because these estimates were largely insensitive to changes in the assumed model. This outcome suggests strong signal in our data. Our analyses support a New Caledonian affiliation of the New Zealand cicada genera Maoricicada, Kikihia, and Rhodopsalta and Australian affinities for the genera Amphipsalta and Notopsalta. This result was surprising, given that previous cicada biologists suspected a close relationship between Amphipsalta, Notopsalta, and Rhodopsalta based on genitalic characters. Relationships among the closely related genera Maoricicada, Kikihia, and Rhodopsalta were poorly resolved, the mitochondrial data and the EF1alpha data favoring different arrangements within this clade. PMID- 11943090 TI - Increased congruence does not necessarily indicate increased phylogenetic accuracy--the behavior of the incongruence length difference test in mixed-model analyses. AB - Comprehensive phylogenetic analyses utilize data from distinct sources, including nuclear, mitochondrial, and plastid molecular sequences and morphology. Such heterogeneous datasets are likely to require distinct models of analysis, given the different histories of mutational biases operating on these characters. The incongruence length difference (ILD) test is increasingly being used to arbitrate between competing models of phylogenetic analysis in cases where multiple data partitions have been collected. Our work suggests that the ILD test is unlikely to be an effective measure of congruence when two datasets differ markedly in size. We show that models that increase the contribution of one data partition over another are likely to increase congruence, as measured by this test. More alarmingly, for many bipartition comparisons, character congruence increases bimodally - either increasing or decreasing the contribution of one data partition will increase congruence - making it impossible to arrive at a single optimally congruent model of analysis. PMID- 11943091 TI - Inferring the root of a phylogenetic tree. AB - Phylogenetic trees can be rooted by a number of criteria. Here, we introduce a Bayesian method for inferring the root of a phylogenetic tree by using one of several criteria: the outgroup, molecular clock, and nonreversible model of DNA substitution. We perform simulation analyses to examine the relative ability of these three criteria to correctly identify the root of the tree. The outgroup and molecular clock criteria were best able to identify the root of the tree, whereas the nonreversible model was able to identify the root only when the substitution process was highly nonreversible. We also examined the performance of the criteria for a tree of four species for which the topology and root position are well supported. Results of the analyses of these data are consistent with the simulation results. PMID- 11943092 TI - Molecular systematics of the Eastern Fence Lizard (Sceloporus undulatus): a comparison of Parsimony, Likelihood, and Bayesian approaches. AB - Phylogenetic analysis of large datasets using complex nucleotide substitution models under a maximum likelihood framework can be computationally infeasible, especially when attempting to infer confidence values by way of nonparametric bootstrapping. Recent developments in phylogenetics suggest the computational burden can be reduced by using Bayesian methods of phylogenetic inference. However, few empirical phylogenetic studies exist that explore the efficiency of Bayesian analysis of large datasets. To this end, we conducted an extensive phylogenetic analysis of the wide-ranging and geographically variable Eastern Fence Lizard (Sceloporus undulatus). Maximum parsimony, maximum likelihood, and Bayesian phylogenetic analyses were performed on a combined mitochondrial DNA dataset (12S and 16S rRNA, ND1 protein-coding gene, and associated tRNA; 3,688 bp total) for 56 populations of S. undulatus (78 total terminals including other S. undulatus group species and outgroups). Maximum parsimony analysis resulted in numerous equally parsimonious trees (82,646 from equally weighted parsimony and 335 from weighted parsimony). The majority rule consensus tree derived from the Bayesian analysis was topologically identical to the single best phylogeny inferred from the maximum likelihood analysis, but required approximately 80% less computational time. The mtDNA data provide strong support for the monophyly of the S. undulatus group and the paraphyly of "S. undulatus" with respect to S. belli, S. cautus, and S. woodi. Parallel evolution of ecomorphs within "S. undulatus" has masked the actual number of species within this group. This evidence, along with convincing patterns of phylogeographic differentiation suggests "S. undulatus" represents at least four lineages that should be recognized as evolutionary species. PMID- 11943093 TI - Delimiting species using DNA and morphological variation and discordant species limits in spiny lizards (Sceloporus). AB - Haplotype phylogenies based on DNA sequence data are increasingly being used to test traditional species-level taxonomies based on morphology. However, few studies have critically compared species limits based on morphological and DNA data, and the methods used to delimit species using either type of data are only rarely explained. In this paper, we review three approaches for species delimitation (tree-based with DNA data and tree-based and character-based with morphological data) and propose explicit protocols for each. We then compare species limits inferred from these approaches, using morphological and mtDNA data for the Yarrow's spiny lizard (Sceloporus jarrovii), a traditionally polytypic species from the southwestern United States and Mexico. All three approaches support division of S. jarrovii into five species, but only two species are the same among the three approaches. We find the greatest support for the five species that are delimited based on mtDNA data, and we argue that mtDNA data may have important (and previously unappreciated) advantages for species delimitation. Because different data and approaches can disagree so extensively, our results demonstrate that the methodology of species delimitation is a critical issue in systematics. PMID- 11943094 TI - A molecular phylogenetic study of the Palmae (Arecaceae) based on atpB, rbcL, and 18S nrDNA sequences. AB - Notoriously slow rates of molecular evolution and convergent evolution among some morphological characters have limited phylogenetic resolution for the palm family (Arecaceae). This study adds nuclear DNA (18S SSU rRNA) and chloroplast DNA (cpDNA; atpB and rbcL) sequence data for 65 genera of palms and characterizes molecular variation for each molecule. Phylogenetic relationships were estimated with maximum likelihood and maximum parsimony techniques for the new data and for previously published molecular data for 45 palm genera. Maximum parsimony analysis was also used to compare molecular and morphological data for 33 palm genera. Incongruence among datasets was detected between cpDNA and 18S data and between molecular and morphological data. Most conflict between nuclear and cpDNA data was associated with the genus Nypa. Several taxa showed relatively long branches with 18S data, but phylogenetic resolution of these taxa was essentially the same for 18S and cpDNA data. Base composition bias for 18S that contributed to erroneous phylogenetic resolution in other taxa did not seem to be present in Palmae. Morphological data were incongruent with all molecular data due to apparent morphological homoplasy for Caryoteae, Ceroxyloideae, Iriarteae, and Thrinacinae. Both cpDNA and nuclear 18S data firmly resolved Caryoteae with Borasseae of Coryphoideae, suggesting that at least some morphological characters used to place Caryoteae in Arecoideae are homoplastic. In this study, increased character sampling seems to be more important than increased taxon sampling; a comparison of the full (65-taxon) and reduced (45- and 33-taxon) datasets suggests little difference in core topology but considerably more nodal support with the increased character sample sizes. These results indicate a general trend toward a stable estimate of phylogenetic relationships for the Palmae. Although the 33-taxon topologies are even better resolved, they lack several critical taxa and are affected by incongruence between molecular and morphological data. As such, a comparison of results from the 45- and 33-taxon trees offers the best available reference for phylogenetic inference on palms. PMID- 11943095 TI - Phylogeny of the Lake Tanganyika cichlid species flock and its relationship to the Central and East African haplochromine cichlid fish faunas. AB - Lake Tanganyika, the oldest of the East African Great Lakes, harbors the ecologically, morphologically, and behaviorally most complex of all assemblages of cichlid fishes, consisting of about 200 described species. The evolutionary old age of the cichlid assemblage, its extreme degree of morphological differentiation, the lack of species with intermediate morphologies, and the rapidity of lineage formation have made evolutionary reconstruction difficult. The number and origin of seeding lineages, particularly the possible contribution of riverine haplochromine cichlids to endemic lacustrine lineages, remains unclear. Our phylogenetic analyses, based on mitochondrial DNA sequences of three gene segments of 49 species (25% of all described species, up to 2,400 bp each), yield robust phylogenies that provide new insights into the Lake Tanganyika adaptive radiation as well as into the origin of the Central- and East-African haplochromine faunas. Our data suggest that eight ancient African lineages may have seeded the Tanganyikan cichlid radiation. One of these seeding lineages, probably comprising substrate spawning Lamprologus-like species, diversified into six lineages that evolved mouthbrooding during the initial stage of the radiation. All analyzed haplochromines from surrounding rivers and lakes seem to have evolved within the radiating Tanganyikan lineages. Thus, our findings contradict the current hypothesis that ancestral riverine haplochromines colonized Lake Tanganyika to give rise to at least part of its spectacular endemic cichlid species assemblage. Instead, the early phases of the Tanganyikan radiation affected Central and East African rivers and lakes. The haplochromines may have evolved in the Tanganyikan basin before the lake became a hydrologically and ecologically closed system and then secondarily colonized surrounding rivers. Apparently, therefore, the current diversity of Central and East African haplochromines represents a relatively young and polyphyletic fauna that evolved from or in parallel to lineages now endemic to Lake Tanganyika. PMID- 11943096 TI - Building supertrees: an empirical assessment using the grass family (Poaceae). AB - Large and comprehensive phylogenetic trees are desirable for studying macroevolutionary processes and for classification purposes. Such trees can be obtained in two different ways. Either the widest possible range of taxa can be sampled and used in a phylogenetic analysis to produce a "big tree," or preexisting topologies can be used to create a supertree. Although large multigene analyses are often favored, combinable data are not always available, and supertrees offer a suitable solution. The most commonly used method of supertree reconstruction, matrix representation with parsimony (MRP), is presented here. We used a combined data set for the Poaceae to (1) assess the differences between an approach that uses combined data and one that uses different MRP modifications based on the character partitions and (2) investigate the advantages and disadvantages of these modifications. Baum and Ragan and Purvis modifications gave similar results. Incorporating bootstrap support associated with pre-existing topologies improved Baum and Ragan modification and its similarity with a combined analysis. Finally, we used the supertree reconstruction approach on 55 published phylogenies to build one of most comprehensive phylogenetic trees published for the grass family including 403 taxa and discuss its strengths and weaknesses in relation to other published hypotheses. PMID- 11943097 TI - Matrix representation with parsimony, taxonomic congruence, and total evidence. PMID- 11943098 TI - Geographic origin of human mitochondrial DNA: accommodating phylogenetic uncertainty and model comparison. AB - Many biogeographic problems are tested on phylogenetic trees. Typically, the uncertainty in the phylogeny is not accommodated when investigating the biogeography of the organisms. Here we present a method that accommodates uncertainty in the phylogenetic trees. Moreover, we describe a simple method for examining the support for competing biogeographic scenarios. We illustrate the method using mitochondrial DNA sequences sampled from modern humans. The geographic origin of modern human mtDNA is inferred to be in Africa, although support for this hypothesis was ambiguous for data from an early paper. PMID- 11943099 TI - Comparing the gap excess ratio and the retention index of the stratigraphic character. PMID- 11943100 TI - A character-based method for measuring the fit of a cladogram to the fossil record. PMID- 11943101 TI - [Stool antigen test for initial Helicobacter pylori diagnosis and for confirmation of eradication after therapy]. AB - BACKGROUND: : Recently, a new diagnostic method based on the detection of Helicobacter pylori antigen in stools (HpSA) has been developed. Our aim was to prospectively evaluate the accuracy of HpSA both in the initial diagnosis of H. pylori infection and in the confirmation of the eradication after treatment. PATIENTS AND METHOD: Thirty dyspeptic patients were prospectively evaluated. During endoscopy, biopsies for histology and rapid urease test were obtained, and a 13C-urea breath test was performed. HpSA was determined by radioimmunoassay(Premier Platinum HpSA). Infected patients were administered H.pylori eradication treatment, and breath test and HpSA were performed again 4 weeks after finishing the therapy. Gold standard for H. pylori infection pre treatment was based on rapid urease test, histology and breath test, whereas 13C urea breath test was considered the post-treatment gold standard. RESULTS: Pre treatment H. pylori prevalence was 57%. Eradication was achieved in 85% of patients. The area under ROC curve for HpSA pre-treatment was 0.98. Cut-off points with best pre-treatment diagnostic accuracy were those between 0.10 and 0.12: positive likelihood ratio (LR), negative LR, 0.06; 94% sensitivity (95% CI, 73-99%),100% specificity (75-100%), 100% positive predictive value (81-100%)and 93% negative predictive value (66-100%). After treatment,the area under ROC curve was 1, and all cut-off points between 0.10 and 0.18 had 100% diagnostic accuracy. HpSA levels decreased from 0.94 (0.9) to 0.08 (0.02) (p < 0.01) in patients withH. pylori eradication success. CONCLUSIONS: HpSA test can be considered an accurate, non-invasive method for the diagnosis of H. pylori infection both inuntreated patients and in the confirmation of H. pylori eradication after treatment. PMID- 11943102 TI - [Helicobacter pylori infection and functional dyspepsia. Meta-analysis of efficacy of eradication therapy]. AB - BACKGROUND: To establish a causal link between Helicobacter pylori infection and functional dyspepsia it is necessary to demonstrate that H. pylori eradication induces an improvement in dyspeptic symptoms. Our aim was to perform a meta analysis of randomized studies comparing, in functional dyspepsia, the efficacy of H. pylori eradication treatment with that of treatments with no effect on H. pylori infection. PATIENTS AND METHODS: DATA SOURCES: PubMed database, Cochrane Controlled Trials Register, and abstracts from congresses until 2001. SELECTION CRITERIA: a) studies including patients with functional dyspepsia and H. pylori infection; b) randomized trials comparing H. pylori eradication treatment with treatment (control) with no effect on H. pylori infection, and c) follow-up of at least 6 months. The quality of studies was assessed by a validated score. STATISTICS: the main outcome was the percentage of patients improving in each therapeutic group. A meta-analysis was performed combining the odds ratios (OR) of individual studies in a global OR. RESULTS: Nine studies fulfilled the inclusion criteria and were thus included in the meta-analysis. Overall, 953 patients received an eradication treatment and 958 received a control treatment. The overall percentage of patients with symptomatic improvement in the eradication group was 43% (95% CI, 40-46%), and it was 39% (95% CI, 36-42%) in the control group. The OR for the effect of the eradication treatment vs. the control treatment was 1.20 (95% CI, 0.91-1.58). The number needed to treat (NNT) with eradication therapy to achieve a symptomatic improvement, compared with the control group, was 25. Although heterogeneity was demonstrated, it disappeared when one study showing positive, clearly discordant results was excluded. Thus, the percentage of patients with symptomatic improvement in the eradication treatment group was 47% (95% CI, 43-50%) and it was 45% (95% CI, 41-48%) in the control group (OR: 1.06; 0.85-1.31; NNT: 50). CONCLUSIONS: H. pylori eradication treatment is not associated with a statistically significant improvement of symptoms in patients with functional dyspepsia. PMID- 11943103 TI - [Surgical treatment of hepatocellular carcinoma. Long term results]. AB - BACKGROUND: Surgical treatment for hepatocellular carcinoma remains controversial due to a lack of prospective randomized studies. MATERIAL AND METHOD: Between January 1990 and December 2000, 121 liver transplantations (group 1) and 52 hepatectomies (group 2) were performed for hepatocellular carcinoma. Each surgical treatment was carried out depending on patients' and tumor's characteristics. RESULTS: Patients from group 1 had a more advanced tumoral grade, with higher involvement of two lobes (19 vs 4%; p = 0.015) and higher number of nodules (1.9 DE [2] vs 1.2 [0.6]; p = 0.001); yet the mean tumor size was lower (3 cm [1.5] vs 4.2 [3.2]; p = 0.006). Operative mortality (4% vs 2%; p = 0.66) and 5- and 10-years survival (68% and 42% vs 63% and 45%; p = 0.23) were similar between both groups. Nevertheless, 5- and 10-years recurrence rates (10.6% and 10.6% vs 50% and 65.5%; p < 0.0001) were more favourable in group 1. Prognostic factors of recurrence included microscopic vascular invasion (RR = 12.12; CI, 2.02-75.52) and alpha-fetoprotein levels higher than 300 ng/mL (RR = 7.12; 95% CI, 1.08-47.02) in group 1, and the pT3-4 stage (RR = 3.86; 95% CI, 1.06-14.03) in group 2. Mean time on waiting lists for liver transplantation was 3.06 (2.66) months and it has increased significantly in last years, especially among blood group 0 patients. However, this fact has not been associated with a worsening of survival rates (p = 0.98). CONCLUSIONS: After a good patient selection, either liver transplantation or hepatectomy achieve excellent long term survival rates in patients with hepatocellular carcinoma, though the former allows a better control of the tumoral disease. The increase of mean time on waiting lists for liver transplantation during the last years has not led to a worsening of survival results. PMID- 11943105 TI - [Transexualism: a challenge for the Spanish National Service]. PMID- 11943104 TI - [First spontaneous reports of adverse reactions to the new selective COX-2 non steroidal anti-inflammatory drugs]. AB - BACKGROUND: [corrected] The new selective cyclooxygenase 2 non-steroidal anti inflammatory drugs (NSAIDs) have been associated with a lower incidence of adverse gastrointestinal effects. MATERIAL AND METHOD: The first Yellow Cards received at the Catalan Centre of Pharmacovigilance reporting adverse effects of rofecoxib and celecoxib were reviewed and compared to those attributed to non selective NSAIDs. RESULTS: Most frequent adverse effects of rofecoxib were gastrointestinal and cardiovascular; those of celecoxib were mostly cutaneous and gastrointestinal; and those of non-selective NSAIDs were gastrointestinal and neuropsychiatric. CONCLUSIONS: Rofecoxib and celecoxib are not devoid of gastrointestinal toxicity; they may also induce cardiovascular adverse effects. PMID- 11943106 TI - [European Resuscitation Council Guidelines 2000 for automated external defibrillation]. PMID- 11943107 TI - [Applicability of flow cytometry in the diagnosis and immunophenotyping follow-up of acute leukemias]. PMID- 11943108 TI - [Anti-obesity drugs: sibutramine and orlistat]. PMID- 11943110 TI - [Drugs and cost-effectiveness]. PMID- 11943112 TI - [Validation of the Spanish version of the Pediatric Asthma Quality of Life Questionnaire in children with asthma]. PMID- 11943113 TI - Predictors of nonsentinel lymph node metastasis in breast cancer patients. AB - BACKGROUND: In order to define a future subset of breast cancer patients in whom the axilla may be staged by sentinel lymph node biopsy alone, the conditions under which nonsentinel axillary lymph node metastases occur must be delineated. METHODS: A prospective database including 212 breast cancer patients who underwent sentinel lymph node biopsy followed by completion axillary dissection at our institution was reviewed. A multivariate, logistic, stepwise regression was performed to evaluate the relationship between nonsentinel lymph node metastasis and patient age, primary tumor size, presence of lymphatic invasion, use of radioisotope to identify the sentinel node and degree of metastasis in the sentinel node. RESULTS: Tumor size greater than 2 cm, lymphatic invasion of the primary tumor, macrometastasis in the sentinel node, and use of radioisotope all positively correlated independently with metastasis in the nonsentinel lymph node (P = 0.0001, P = 0.0483, P = 0.0008, P = 0.0271, respectively). CONCLUSIONS: Predictors of nonsentinel axillary node metastasis exist and are important in defining those patients in whom a sentinel lymph node biopsy alone may not be adequate. PMID- 11943114 TI - Immediate reconstruction after mastectomy for breast cancer does not prolong the time to starting adjuvant chemotherapy. AB - BACKGROUND: Immediate breast reconstruction is often performed after mastectomy for breast cancer. There has been concern that this will result in a delay in initiating chemotherapy and, as a consequence, may adversely impact survival. In this study we sought to determine whether immediate breast reconstruction affects the interval between surgery and adjuvant chemotherapy. METHODS: A single institution retrospective analysis was made using the institutional tumor registry and chart reviews. RESULTS: Forty-nine patients were identified who had undergone mastectomy with immediate reconstruction followed by adjuvant chemotherapy. They were compared with 308 patients undergoing mastectomy without reconstruction. Patients who underwent reconstruction were overall younger (46 versus 55, P <0.001), and had more advanced disease. The time to chemotherapy was significantly longer in the group receiving no reconstruction: 53 versus 41 days (P = 0.039). The type of reconstruction did not affect the time to chemotherapy. CONCLUSIONS: Immediate reconstruction after mastectomy does not increase the time to chemotherapy compared with mastectomy alone. PMID- 11943115 TI - The impact of practice setting and financial incentives on career satisfaction and perceived practice limitations among surgeons. AB - OBJECTIVE: To study how practice setting and financial incentives affect career satisfaction and perceived impact on practice in surgery. METHODS: Data are from the Community Tracking Study (CTS) physician survey, a national survey of active physicians in the United States fielded between August 1996 and August 1997. Surveys with 1,738 practicing surgeons were completed. To be eligible, surgeons had to have completed their medical training (which excludes residents, interns, or fellows), be practicing in the contiguous United States, and be providing direct patient care for at least 20 hours per week. The dependent variables measured are career dissatisfaction and perceived limitations/pressures on time spent with patients, clinical freedom, income, and continuity. The dependent variables are regressed on practice setting, percentage of managed care practice revenue, individual financial incentives, age group, gender, international medical graduate, and board certification. RESULTS: Working in a small practice is the strongest predictor of career dissatisfaction, with about twice the adjusted rate of career dissatisfaction (26%) than other practice settings (13% in group practices, 16% in staff-model HMOs, 9% in medical schools, 18% in hospitals). Managed care plays a much smaller role; an increase in the dependence on managed care equivalent to the difference between 25th and 75th percentile only increases career dissatisfaction by 3 percentage points. Surgeons in solo or two physician practices are also more likely than surgeons in other settings to report that income pressure and limitations on clinical freedom and patient continuity compromise quality of care. PMID- 11943116 TI - Early esophageal transit study after laparoscopic fundoplication: how useful is it? AB - BACKGROUND: Early complications of laparoscopic fundoplication, if immediately recognized, may be promptly treated laparoscopically with minimal morbidity. A suggested strategy for identification is a routine postoperative esophageal transit study. OBJECTIVE: To investigate the role of early postoperative esophagogram with Gastrografin in predicting major complications, failures, or severe dysphagia. DESIGN: Esophagograms performed in 92 patients, 24 hours after laparoscopic fundoplication, were correlated to major complications. Esophageal transit time was scored and correlated with dysphagia. RESULTS: Esophagogram detected two of three observed complications: acute paraesophageal hernia and intrathoracic migration, but not a fundic perforation. Only a severe transit impairment predicted a disabling dysphagia (specificity 82%, sensitivity 70%). CONCLUSIONS: Postoperative swallow is an appropriate investigation to diagnose anatomical abnormalities but may be deceptive for perforations. Severe transit delay may predict the risk of severe dysphagia. Although useful, postoperative routine transit studies would probably not change the therapeutic strategies in most patients. PMID- 11943117 TI - How early is early laparoscopic treatment of acute cholecystitis? AB - BACKGROUND: Despite the well-accepted success of laparoscopic cholecystectomy in elective treatment of symptomatic cholelithiasis, the efficacy and timing of this technique has been subject to some debate in the setting of acute cholecystitis. This study was undertaken to evaluate our institution's experience with early cholecystectomy as a safe, effective treatment of acute cholecystitis. METHODS: Charts of all patients who had undergone laparoscopic cholecystectomy for the diagnosis of acute cholecystitis were reviewed. Patients were divided into two groups based on the length of time from onset of symptoms to surgical intervention: less than 48 hours in the early group (n = 14) and more than 48 hours in the late group (n = 31). RESULTS: Comparing the two groups, the conversion rate to an open procedure was significantly less (0 versus 29%, P <0.04) in the early treated patients. Furthermore, the operative time (73 versus 96 minutes, P <0.004), postoperative hospitalization (1.2 versus 3.9 days, P <0.001), and total hospital stay (2.1 versus 5.4 days, P <0.004) were significantly reduced in patients undergoing early laparoscopic cholecystectomy. CONCLUSIONS: Laparoscopic cholecystectomy performed by experienced surgeons is a safe, effective technique for treatment of acute cholecystitis. Patients treated within 48 hours of onset of symptoms experience a lower conversion rate to an open procedure, shorter operative time and reduced hospitalization. PMID- 11943118 TI - Morbidity, mortality, and technical factors of distal pancreatectomy. AB - BACKGROUND: Pancreatic leak is a major source of morbidity associated with pancreatic surgery. We sought to identify disease and technique-dependent factors associated with morbidity and mortality after distal pancreatectomy. METHODS: Retrospective review of patients who underwent distal pancreatectomy during a 5 year period. Clinical, technical, and pathologic data were correlated with operative morbidity or mortality. RESULTS: Fifty-one patients underwent distal pancreatectomy for primary pancreatic disease, extrapancreatic malignancy, or trauma. Overall perioperative mortality and morbidity rates were 4% and 47%, respectively. Pancreatic leak was the most common complication, occurring in 26% of patients. Overall complications and pancreatic leaks occurred more often after distal pancreatectomy for trauma and in patients with a sutured pancreatic stump closure. CONCLUSIONS: Distal pancreatectomy can be performed with a low rate of mortality, though pancreatic leak is a common cause of morbidity. The urgency of the procedure and the method of pancreatic stump closure may influence postoperative morbidity. PMID- 11943119 TI - A role for error training in surgical technical skill instruction and evaluation. AB - BACKGROUND: During the evaluation of many instances of the same basic surgical skill, we observed that there were several errors that occurred frequently. Two studies were undertaken to examine the use of these errors for improving the instruction and evaluation of the skill. MATERIALS AND METHODS: For both studies, two types of rater training videotapes were developed. One involved the use of examples of common errors (error) and the other demonstrated the skill being performed correctly (correct). A testing videotape was created consisting of 24 performances of the skill that ranged in quality of the performance. The first study was designed to assess the impact of error instruction on skill acquisition. In this study, a group of 30 senior medical students were randomly assigned to one of four different training groups: none, error only, correct only, and error+correct. Subjects were videotaped performing the skill before and after the training and three experts evaluated these performances independently using a 7-point rating scale. The second study was designed to assess the impact of error training on skill evaluation and was done using both novice and expert raters. The same group of 30 senior medical students used in the first study was used as novice raters. Following training in one of the four training groups, each subject rated the 24 performances on the testing videotape and interrater reliability was assessed for each group. Surgical faculty served as expert raters in this study and were randomly assigned to receive either error training or no training. Each subject viewed the testing videotape, rating the performances and giving "feedback" commentary. Interrater reliability was calculated for the two groups and the precision of the feedback was assessed. RESULTS: Significant improvement in posttest performance scores was seen only in the "error+correct" training group. Interrater reliability was somewhat lower for the "correct only" and "error only" training groups in both the student and faculty studies. Faculty raters receiving error training had a higher proportion of specific comments than the group that received no training although this difference was not statistically significant. CONCLUSIONS: Instruction about common errors, when combined with instruction about the correct performance enhanced the acquisition of this surgical skill. This suggests a role for the use of errors in surgical technical skill instruction. Our study provides no support for a role for error training in improving skill evaluation. PMID- 11943120 TI - A critical evaluation of the morbidity and mortality conference. AB - BACKGROUND: This study was designed to evaluate the impact of changes made to our morbidity and mortality (M&M) conference. METHODS: A 23-item survey using corresponding Likert-type scales was created. Faculty and residents were asked to anonymously complete the surveys in June 1999. Based on this information, specific modifications were made to the conference. The same survey was administered to faculty and residents in the Fall of 2000. Analysis was performed using Student t tests. RESULTS: Postsurvey findings showed residents felt eight components improved significantly (P <0.05). Faculty noted nonsignificant improvement in nine survey items and decline in nine items (five unchanged). CONCLUSIONS: Changes in content and structure made to enhance our M&M conference's educational value resulted in significant improvements as perceived by the surgical residents. Interestingly, these changes had only minimal impact on faculty perceptions. PMID- 11943121 TI - Identification of teaching excellence in operating room and clinic settings. AB - BACKGROUND: A system for obtaining learner feedback on surgical faculty teaching is a program-specific resource for recognizing faculty accomplishments as well as being a requirement of the Accreditation Council for Graduate Medical Education (ACGME). This investigation uses 5 years of feedback from residents to identify surgical teaching behaviors that define teaching excellence. METHODS: Between 1995 and 1999 full-time surgeons in a division of general surgery were evaluated biannually by every resident on their services, using two 10-item Likert scales to assess frequency of performing selected teaching behaviors. Response categories ranged from 0 (does not demonstrate) to 4 (demonstrates the behavior to a very high degree). Mean scores > or =3.7 (1 SD above the mean) were categorized as evidence of superior teaching, whereas mean scores < or =2.4 (1 SD below the mean) were categorized as mediocre. Residents wrote statements identifying teaching strengths. RESULTS: There were 753 individual resident assessments of 16 faculty. The overall mean rating for operating room and clinic teaching was 3.1, with 24% of the ratings > or =3.7 and 14% of the ratings < or =2.4. For operating room, discriminant behaviors were: demonstrates sensitivity to resident learning needs (3.85 versus 1.62, P <0.01) and provides direct feedback (3.60 versus 1.27, P <0.01). Residents' statements yielded themes tied to superior teaching: demonstrates technical expertise, allows resident participation, and maintains a learning climate of respect. CONCLUSIONS: A resident-based teaching assessment system can offer a reasonable and valid form of feedback to academic surgeons. The use of mixed methods to identify teaching behaviors that characterize excellence informs faculty of how they are perceived as educators and provides examples of specific behaviors that merit commendation. PMID- 11943122 TI - Emergency operations for nondiverticular perforation of the left colon. AB - OBJECTIVE: Although diverticulitis is the most common cause of large bowel perforation, other disease may result in left colonic peritonitis. The aim of this study was to evaluate and compare the incidence, management, and outcome of patients with different causes of nondiverticular left colonic perforations. PATIENTS AND METHODS: From January 1992 to September 2000, 212 surgical patients underwent emergency operation for distal colonic peritonitis. Perforations were caused by diverticulitis in 133 patients (63%) and by a nondiverticular process in 79 (37%). Mortality and morbidity in patients with nondiverticular perforation of the distal large bowel its relationship with the general conditions, the grade and the cause of peritonitis were analysed. Four types of surgical procedures were used. Hartmann's procedure was performed in 40 patients (51%); intraoperative colonic lavage, resection, and primary anastomosis (ICL) in 27 patients (34%); colostomy in 7 (9%); and subtotal colectomy in 5 (6%). RESULTS: Perforated neoplasm, the most common cause of peritonitis, was observed in 30 patients, colonic ischemia in 20, iatrogenia in 13, and other causes in 16 patients. One or more complications were observed in 57 patients (72%); among causes of perforation, colonic ischemia was significantly associated with the longest hospital stay and highest mortality. Eighteen patients (23%) died. CONCLUSIONS: Left large bowel perforation by nondiverticular disease is associated with high mortality and morbidity. The prognosis of patients is determined by the development of septic shock and colonic ischemia, as underlying disease, may influence patient survival. PMID- 11943123 TI - When should we operate on major fractures in patients with severe head injuries? AB - BACKGROUND: The widely accepted practice of early fracture fixation (EFF) in multiply injured patients has recently been challenged in the presence of head injury. DATA SOURCES: English and German language articles on the subject were searched using Medline. Keywords included head trauma, intracranial trauma, brain injuries, fractures, fracture fixation, timing, femur fracture, and tibia fracture. CONCLUSIONS: The available literature does not provide clear-cut guidance on the management of fractures in the presence of head injuries. The trend is toward a better outcome if the fractures are fixed early. Treatment should therefore be tailored to the individual patient, with the assumption that full neurologic recovery will take place. PMID- 11943124 TI - Cytokines for surgeons. AB - All cells maintain continuous communication. Hormones derive constitutively from specialized cells to effect total body homeostasis. Conversely, cytokines are produced sporadically from almost all nucleated cells in response to surgical ischemia/septic challenge. Surgical patients are a stew of pulsating cytokines, which serve as the language between all surgically stressed somatic and myeloid cells. Therapeutic manipulation of cytokines has already generated some exhilarating success stories and some crushing disappointments. This introduction to surgically relevant cytokines is presented with the conviction that cytokine based therapies of surgical patients will (in the future) prove as beneficial to our patients as antibiotics have in the past. PMID- 11943125 TI - A meta-analysis of randomized controlled trials that compared neoadjuvant chemotherapy and surgery to surgery alone for resectable esophageal cancer. AB - BACKGROUND: Esophagectomy is often considered the standard treatment for resectable esophageal cancer but the rate of cure is low. Combining neoadjuvant chemotherapy with surgery has theoretical appeal and some clinical evidence suggests a benefit. We performed a meta-analysis of randomized controlled trials (RCTs) that compared neoadjuvant chemotherapy and surgery with surgery alone for esophageal cancer. METHODS: Medline and manual searches were done to identify all published RCTs that compared neoadjuvant chemotherapy and surgery to surgery alone for esophageal cancer. The selection process was inclusive; no trials were excluded. Trial validity assessment was done and a trial quality score was assigned. Outcomes assessed by meta-analysis included 1-, 2-, and 3-year survival, rate of resection, rate of complete resection, operative mortality, anastomotic leaks, postoperative pulmonary complications, all treatment mortality, local-regional cancer recurrence, distant cancer recurrence, and all cancer recurrence. A random-effects model was used and odds ratio was the principal measure of effect. Systematic quantitative review was done for outcomes unique to the neoadjuvant chemotherapy treatment group (clinical response, pathological complete response, and chemotherapy mortality). RESULTS: Eleven RCTs, which included 1,976 patients, were selected with quality scores ranging from 1 to 3 (5-point Jadad scale). Odds ratio (95% confidence interval [CI]; P value), expressed as chemotherapy and surgery versus surgery alone (treatment versus control; values <1 favor chemotherapy-surgery arm), was 1.00 (0.76, 1.30; P = 0.98) for 1-year survival, 0.88 (0.62, 1.24; P = 0.45) for 2-year survival, 0.77 (0.37, 1.59; P = 0.48) for 3-year survival, 1.71 (1.22, 2.40; P = 0.002) for rate of resection, 0.71 (0.58, 0.87; P = 0.001) for rate of complete resection, 0.94 (0.66, 1.35; P = 0.76) for operative mortality, 1.08 (0.45, 2.60; P = 0.87) for anastomotic leaks, 1.31 (0.77, 2.23; P = 0.32) for postoperative pulmonary complications, 1.36 (0.83, 2.25; P = 0.22) for all treatment mortality, 0.71 (0.36, 1.42; P = 0.33) for local-regional cancer recurrence, 0.79 (0.57, 1.10; P = 0.16) for distant cancer recurrence, and 0.63 (0.28, 1.41; P = 0.26) for all cancer recurrence. A clinical response to chemotherapy was observed in 31% of patients and 5% had a complete pathological response. Chemotherapy mortality (before surgery) was 1.6%. CONCLUSIONS: Compared with surgery alone, neoadjuvant chemotherapy and surgery is associated with a lower rate of esophageal resection but a higher rate of complete (R0) resection. It does not increase treatment related mortality. This meta-analysis did not demonstrate a survival benefit for the combination of neoadjuvant chemotherapy and surgery. PMID- 11943127 TI - Binding pancreaticojejunostomy is a new technique to minimize leakage. AB - Pancreaticoduodenectomy (Whipple procedure) has been the standard treatment for periampullary and pancreatic carcinoma. A leak or fistula from the pancreatic anastomosis is the leading cause of morbidity and mortality after pancreaticoduodenectomy. In order to effectively prevent the development of pancreatic fistulae, we designed a special technique called binding pancreaticojejunostomy, by which 3 cm of the serosa-muscular sheath of the jejunum was bound to the pancreatic remnant. We have performed this procedure in 105 consecutive patients; none of the cases developed pancreatic fistula. It is a safe, simple, and efficient technique. PMID- 11943126 TI - Evaluation of a new technique for bedside percutaneous tracheostomy. AB - BACKGROUND: Percutaneous tracheostomy as described by Ciaglia is accepted as a safe technique with minimal associated morbidity. Recent modification of the technique to a single-step dilator prompted us to evaluate this in the critically injured patient. METHODS: A comparison of patients undergoing percutaneous tracheostomy was performed. From May 1998 to May 1999, patients underwent surgery using the sequential multidilator technique (MDT), and from July 1999 to July 2000, patients underwent surgery using the single dilation technique (SDT). RESULTS: Ninety-three tracheostomies were performed, 49 MDT and 44 SDT. Time to tracheostomy and total ventilator days was similar between the groups. Three complications occurred. In the MDT group, 1 patient experienced delayed tracheal hemorrhage not requiring transfusion. In the SDT group, 1 patient had transient right lower lobe collapse, and another patient had unexplained extubation requiring emergent cricothyroidotomy. CONCLUSIONS: Percutaneous tracheostomy using the single-step Rhino dilator technique is technically easier than the currently accepted multidilator technique with equivalent complications. PMID- 11943128 TI - Video-assisted thyroid lobectomy through a small wound in the submandibular area. AB - BACKGROUND: Endoscopic thyroidectomy has not gained wide acceptance because of the expertise required, the long operation time, the wide dissection, and the extra cost of specialized instruments. We developed a video-assisted hemithyroidectomy procedure that requires only one small incision at the upper neck. METHODS: Hemithyroidectomy was performed through a 25 to 30 mm transverse incision made in the upper lateral neck for the treatment of benign thyroid nodule. No gas or external lift dissection was needed. RESULTS: The mean age of 39 patients was 33.8 years. The tumor size ranged from 1.9 to 5.5 cm (mean 3.1 cm). All patients underwent total lobectomy without conversion to traditional cervicotomy. The mean operation time was 56 minutes (range 36 to 90). Follicular adenoma was the final pathologic diagnosis in 25 patients and adenomatous goiter in 14. Transient recurrent laryngeal nerve palsy was seen in 1 patient. CONCLUSIONS: Our technique is safe, minimally invasive, less time consuming, and cosmetically excellent. PMID- 11943129 TI - Intestinal tuberculosis presenting as a bowel obstruction. PMID- 11943130 TI - Surgical management of renal cell carcinoma with inferior vena cava tumor thrombus. AB - BACKGROUND: The successful excision of a renal cell carcinoma (RCC) invading the inferior vena cava (IVC) remains a technical intraoperative challenge and requires a careful preoperative surgical management planning. Although a radical operation remains the mainstay of the therapy for RCC, the optimal management of the patients with RCC causing IVC tumor thrombus remains unresolved. In this study, we reviewed our experience in this group of patients and herein report the results. METHODS: Between July 1990 and August 1998, 11 patients with RCC with IVC tumor thrombus underwent surgical treatment. The mean patient age was 54.2 years and the male to female ratio was 1.75. The cephalad extension of the tumor was suprarenal in all cases, being infrahepatic in 6 patients, intrahepatic in 2, and suprahepatic with right atrial extension in 3 patients. All tumors were resected via inferior vena cava isolation and, when necessary, extended hepatic mobilization and Pringle maneuver, with primary or patch closure of vena cavotomy. Cardiopulmonary bypass (CPB) and deep hypothermic circulatory arrest (DHCA) were used in 3 patients. RESULTS: The mortality rate was 9.1% (1 patient was lost on the 11th postoperative day). Complications occurred in 3 patients. The remaining 10 patients (90.9%) could be successfully discharged from hospital. Two of them were lost during follow-up because of tumor progression at the 43rd and 54th postoperative months. The 10-year Kaplan-Meier survival estimate was 71.4%, with a mean follow-up of 4.6 years. The presence of lymph node metastases and perinephric spread seemed to possess an adverse effect on the survival. Although the groups included small numbers of patients, there was no significant difference in survival in regard to the different levels of tumor thrombus extension into the vena cava. CONCLUSIONS: Surgical treatment is the preferred approach to patients with RCC and IVC tumor thrombi as it provides markedly better results when compared with the other therapeutical modalities. We believe that complete surgical excision of the tumor and the resulting thrombus with appropriate preoperative staging and a well-planned surgical approach, using CPB and DHCA when necessary, provide an acceptable long-term survival with a good quality of life expectation. PMID- 11943131 TI - An encounter card system for increasing feedback to students. AB - BACKGROUND: Student satisfaction with the feedback process is essential for motivating students to improve their performance. METHODS: Third-year medical students participated in a system to increase feedback from faculty, fellows, and residents during a 12-week surgery clerkship. Each student received 40 encounter cards to solicit verbal and written feedback on specific domains of clinical performance. The clerkship evaluations from the intervention group (Fall 2000 2001) were compared with a historical control group (Fall 1999-2000) on seven feedback-related items. Effect sizes were calculated and two-tailed t tests performed to determine statistical significance. RESULTS: All seven items had effect sizes greater than 0.30 (range, 0.38 to 0.85). Differences between the two groups' scores were statistically significant for six items. CONCLUSIONS: Implementation of an encounter card system improved student satisfaction with the feedback process. Intervention group students perceived that faculty and residents had improved their understanding of history-taking, physical examination, and decision-making skills. PMID- 11943132 TI - The development of an interactive game-based tool for learning surgical management algorithms via computer. AB - BACKGROUND: We have previously demonstrated the potential efficacy of a computer assisted board game as a tool for medical education. The next logical step was to transfer the entire game on to the computer, thus increasing accessibility to students and allowing for a richer and more accurate simulation of patient scenarios. METHODS: First, a general game model was developed using Microsoft Visual Basic. A breast module was then created using 3-D models, radiographs, and pathology and cytology images. The game was further improved by the addition of an animated facilitator, who directs the players via gestures and speech. Thirty three students played the breast module in a variety of team configurations. After playing the game, the students completed surveys regarding its value as both an educational tool and as a form of entertainment. 10-question tests were also administered before and after playing the game, as a preliminary investigation into its impact on student learning. RESULTS: After playing the game, mean test scores increased from 6.43 (SEM +/- 0.30) to 7.14 (SEM +/- 0.30; P = 0.006). The results of the five-question survey were extremely positive. Students generally agreed that the game concept has value in increasing general knowledge regarding the subject matter of breast disease and that the idea of following simultaneously the work-up of numerous patients with similar problems is a helpful way to learn a work-up algorithm. CONCLUSIONS: Postgame surveys demonstrate the efficacy of our computer game model as a tool for surgical education. The game is an example of problem based learning because it provides students with an initial set of problems and requires them to collect information and reason on their own in order to solve the problems. Individual game modules can be developed to cover material from different diagnostic areas. PMID- 11943133 TI - Current status of liver transplantation for hepatocellular cancer. AB - The incidence of hepatocellular cancer is increasing in the United States and is one of the most common cancers worldwide. Traditionally, the gold standard treatment for hepatocellular cancer has been surgical resection, but most patients were not suitable candidates due to advanced disease. Other treatments include locally ablative techniques (cryosurgery, radiofrequency ablation and various injection therapies), chemotherapeutic options and rarely, radiation therapies. In the 1980s, liver transplant emerged as the treatment of choice for end-stage liver disease and also became an option for patients with hepatocellular cancer. When comparing liver transplant with resection in retrospective studies, liver transplant patients had better survival and reduced recurrence. However, not all patients with hepatocellular cancer will be candidates for liver transplant. Size, stage, and histological grade of tumor all affect prognosis after transplant. Use of chemotherapeutic treatments and locally ablative techniques may be beneficial prior to liver transplant, but larger controlled studies are needed. Liver transplant is the most effective treatment for hepatocellular cancer in the subgroup of smaller tumors, but ultimately we are limited by the number of available donors. Future goals in this area include increasing the donor pool and determining optimal management to allow patients to wait for an appropriate donor. PMID- 11943135 TI - Chromatin remodeling during spermiogenesis: a possible role for the transition proteins in DNA strand break repair. AB - An important chromatin remodeling process is taking place during spermiogenesis in mammals and DNA strand breaks must be produced to allow the accompanying change in DNA topology. Endogenous DNA strand breaks are indeed detected at mid spermiogenesis steps but are no longer present in mature sperm. Both in vitro and in vivo evidence suggests that the DNA-binding and condensing activities of a set of basic nuclear "transition proteins" may be crucial to the integrity of the chromatin remodeling process. We propose that these proteins are necessary for the repair of the strand breaks so that DNA fragmentation is minimized in the mature sperm. PMID- 11943134 TI - Gastropyloric motor activity and the effects of erythromycin given orally after esophagectomy. AB - BACKGROUND: The motor activity of the gastric tube as an esophageal replacement after esophagectomy is poorly understood. The aims of the present study were to examine the gastropyloric motility of the gastric tube and the effects of erythromycin given orally. METHODS: Interdigestive gastropyloric motility was recorded by manometry with a sleeve sensor in 23 esophagectomized patients. The 23 patients were classified into 3-, 12-, and 24-month groups according to postoperative follow-up time. Radiopaque markers were used in 8 patients to assess gastric emptying. The effects of erythromycin were studied after the patients received 600 mg during fasting and 1 g postprandially. RESULTS: Compared with the 3-month group, the 12-month group and the 24-month group showed significantly increased pyloric and antral motility, respectively. During a fast, erythromycin induced phase III in 44.4% of the patients with more than 12 months of follow-up. In contrast to the normal subjects, esophagectomized patients showed delayed gastric emptying at 3 and 4 hours. However, erythromycin significantly accelerated gastric emptying at 1, 2, 3, and 4 hours. CONCLUSIONS: The motor activity of the gastric tube returns towards normal in a progression over time from the pylorus cephalad. Erythromycin given orally might be used as a prokinetic agent in patients after esophagectomy. PMID- 11943136 TI - The allograft inflammatory factor-1 family of proteins. AB - The allograft inflammatory factor-1 (AIF-1) is a 17 kDa interferon-gamma inducible Ca(2+)-binding EF-hand protein that is encoded within the HLA class III genomic region. Three proteins are probably identical with AIF-1 termed Iba1 (ionized Ca(2+)-binding adapter), MRF-1 (microglia response factor) and daintain. Considerable but not complete sequence identity with AIF-1 has been described for IRT-1 (interferon-responsive transcript), BART-1 (balloon angioplasty-responsive transcript), and other, yet unassigned alternatively spliced variants. In this review, genomic and functional characteristics of AIF-1-related proteins are summarized and a common nomenclature is proposed. PMID- 11943138 TI - Genome-wide detection and family clustering of ion channels. AB - Ion channels represent an important class of molecules that can be classified into 13 distinct groups. We present a strategy using a "learning set" of well annotated ion channel sequences to detect homologues in 32 entire genome sequences from Archaea, Bacteria and Eukarya. A total of 299 putative ion channel protein sequences were detected, with significant variations across species. The clustering of these sequences reveals complex relationships between the different ion channel families. PMID- 11943139 TI - Evidence for a functional nitric oxide synthase system in brown adipocyte nucleus. AB - The intracellular localization and activity of the nitric oxide synthase (NOS) isoforms were investigated in rat brown adipocytes. Immunohistochemistry showed cytoplasmic and nuclear staining for the endothelial NOS (eNOS) and inducible NOS (iNOS) isoforms; accordingly, anti-L-citrulline antibody, a marker of NOS activity, immunostained both the cytoplasm and the nucleus. The presence of metabolically active NOS in the nucleus was further confirmed by immunoblotting analyses of subcellular fractions of homogenates from cultured brown adipocytes and by measurements of NOS activity in the cytosol and nucleus. Sympathetic stimulation in vivo (i.e. cold exposure or beta(3)-adrenergic agonist treatment) and in vitro (i.e. noradrenaline treatment of cultured cells) significantly increased both cytosolic and nuclear eNOS and iNOS expression and activities. By contrast, the number of iNOS-positive, but not eNOS-positive, nuclei was significantly lower in the functionally impaired brown fat of genetically obese Zucker fa/fa rats. These data suggest the existence of a noradrenaline-modulated functional NOS system in the nucleus of brown adipocytes. PMID- 11943137 TI - Coupling endoplasmic reticulum stress to the cell death program: role of the ER chaperone GRP78. AB - Alterations in Ca(2+) homeostasis and accumulation of unfolded proteins in the endoplasmic reticulum (ER) lead to an ER stress response. Prolonged ER stress may lead to cell death. Glucose-regulated protein (GRP) 78 (Bip) is an ER lumen protein whose expression is induced during ER stress. GRP78 is involved in polypeptide translocation across the ER membrane, and also acts as an apoptotic regulator by protecting the host cell against ER stress-induced cell death, although the mechanism by which GRP78 exerts its cytoprotective effect is not understood. The present study was carried out to determine whether one of the mechanisms of cell death inhibition by GRP78 involves inhibition of caspase activation. Our studies indicate that treatment of cells with ER stress inducers causes GRP78 to redistribute from the ER lumen with subpopulations existing in the cytosol and as an ER transmembrane protein. GRP78 inhibits cytochrome c mediated caspase activation in a cell-free system, and expression of GRP78 blocks both caspase activation and caspase-mediated cell death. GRP78 forms a complex with caspase-7 and -12 and prevents release of caspase-12 from the ER. Addition of (d)ATP dissociates this complex and may facilitate movement of caspase-12 into the cytoplasm to set in motion the cytosolic component of the ER stress-induced apoptotic cascade. These results define a novel protective role for GRP78 in preventing ER stress-induced cell death. PMID- 11943140 TI - Exploring the subsite specificity of Schistosoma mansoni aspartyl hemoglobinase through comparative molecular modelling. AB - Blood flukes of the genus Schistosoma currently infect millions of people in tropical and subtropical countries. An enzyme playing a major role in hemoglobin (Hb) degradation by Schistosoma mansoni has been cloned and shown to be highly similar to the human cathepsin D aspartyl proteinase, although presenting a distinct substrate specificity from the latter. Investigating the structural features responsible for this difference has a major application in the design of selective anti-schistosomal drugs. In order to achieve this goal a homology model for the S. mansoni aspartyl hemoglobinase was constructed and then used to simulate the complexes formed with two transition state analogues of Hb-derived octapeptide substrates. Comparison with human cathepsin D showed that different pocket volumes and surface electrostatic potentials arise from substitutions in residues comprising the S4, S3, S2 and S3' subsites. Since the primary specificity of the S. mansoni enzyme resembles that of HIV-1 protease, we have discussed the applicability of current retroviral protease inhibitors as leads for the design of new anti-schistosomal drugs. PMID- 11943141 TI - The major light-harvesting pigment protein of Acaryochloris marina. AB - The major light-harvesting protein complex containing chlorophyll (Chl) d was isolated from Acaryochloris marina thylakoid membranes. Isolation was achieved by detergent solubilisation followed by separation on 6-40% sucrose gradients using ultracentrifugation. The best Chl d yield (70%) used 0.3% dodecyl maltoside, 0.15% octyl glucoside, 0.05% zwittergent 3-14 with the detergent:total Chl d ratio around 10:1 (w/w). Characterisation of the light-harvesting pigment protein complex (lhc) involved non-denaturing electrophoresis, SDS-PAGE, absorbance and fluorescence spectroscopy. The main polypeptide in the lhc was shown to be ca. 34 kDa and to contain Chl d and Chl a, indicating that the Acaryochloris lhc is similar to that of prochlorophytes. The Chl a level varied with the culture conditions, which is consistent with previous findings. PMID- 11943142 TI - Vav-induced activation of the human IFN-gamma gene promoter is mediated by upregulation of AP-1 activity. AB - The role of Vav in the transcriptional regulation of the human interferon-gamma (IFN-gamma) promoter was investigated. Overexpression of Vav in Jurkat-TAg cells enhanced T cell receptor (TCR)-induced activation of a luciferase (Luc) reporter gene construct driven by cis-regulatory element of the IFN-gamma gene (-346 to +7). Electrophoresis mobility shift and Luc reporter assays demonstrated that the DNA-binding and transcriptional activity of the proximal AP-1-dependent NFAT site (positions -172 to -138), the AP-1/Ying-Yang 1 (YY1)-binding site (-209 to -184), and a consensus AP-1-binding site were upregulated by Vav. Vav enhanced TCR induced activation of c-Jun N-terminal kinase (JNK) and its upstream regulator, Rho family GTPases. Finally, coexpression of a dominant-negative Rac1 mutant suppressed Vav-mediated upregulation of the transcriptional and DNA-binding activity of the proximal NFAT/AP-1 site and the AP-1/YY1 site, as well as the complete IFN-gamma promoter activity. Vav activates the IFN-gamma promoter via upregulation of AP-1-binding through a Rac1/JNK pathway. PMID- 11943143 TI - PrP-dependent cell adhesion in N2a neuroblastoma cells. AB - The cellular isoform of prion protein (PrP(C)) is a ubiquitous glycoprotein expressed by most tissues and with a biological function yet to be determined. Here, we have used a neuroblastoma cell model to investigate the involvement of PrP in cell adhesion. Incubation of single cell suspension induced cell-cell adhesion and formation of cell aggregates. Interestingly, cells overexpressing PrP exhibit increased cation-independent aggregation. Aggregation was reduced after phosphatidylinositol-specific phospholipase C release of the protein and by pre-incubation of cells with an antibody raised against the N-terminal part of PrP(C). Our paradigm allows the study of the function of PrP as an intercellular adhesion molecule and a cell surface ligand or receptor. PMID- 11943144 TI - The identification of the acid-base catalyst of alpha-arabinofuranosidase from Geobacillus stearothermophilus T-6, a family 51 glycoside hydrolase. AB - The alpha-L-arabinofuranosidase from Geobacillus stearothermophilus T-6 (AbfA T 6) belongs to the retaining family 51 glycoside hydrolases. The conserved Glu175 was proposed to be the acid-base catalytic residue. AbfA T-6 exhibits residual activity towards aryl beta-D-xylopyranosides. This phenomenon was used to examine the catalytic properties of the putative acid-base mutant E175A. Data from kinetic experiments, pH profiles, azide rescue, and the identification of the xylopyranosyl azide product provide firm support to the assignment of Glu175 as the acid-base catalyst of AbfA T-6. PMID- 11943145 TI - The acidic regions of WASp and N-WASP can synergize with CDC42Hs and Rac1 to induce filopodia and lamellipodia. AB - The acidic (A) region of WASp family proteins is thought to represent a high affinity binding site for Arp2/3 complex without activating properties. Here we show that GST-fused WASp-A and N-WASP-A, but not a WASP-A/W500S mutant, several truncated WASp-A constructs or WAVE1-A can pull down Arp2/3 complex from cell lysates. Significantly, WASp-A and N-WASP-A synergistically trigger formation of filopodia or lamellipodia when coinjected with sub-effective concentrations of V12CDC42Hs or V12Rac1, respectively, into macrophages. The ability of WASp family A region constructs to induce this effect is closely correlated with their ability to bind Arp2/3 complex in vitro. These results imply that (i) Arp2/3 complex is critically involved in filopodia and lamellipodia formation in macrophages and (ii) acidic regions of WASp and N-WASP are not simply binding sites for Arp2/3 complex but can prime it for RhoGTPase-triggered signals leading to actin nucleation. PMID- 11943146 TI - Ligand-induced shedding of discoidin domain receptor 1. AB - Tyrosine kinases belonging to the discoidin domain receptor (DDR) family are activated upon stimulation with various types of collagen. In response to collagen treatment, immunoprecipitation of DDR1 with an antibody specific to the juxtamembrane region results in co-purification of a previously unrecognized tyrosine phosphorylated protein of 62 kDa molecular weight. Here, this protein is identified as C-terminal cleavage product of the full-length DDR1 receptor and a DDR1-specific shedding enzyme postulated. Shedding of DDR1 can be partially blocked by the furin inhibitor decanoyl-RVKR-chloromethylketone and completely inhibited by the hydroxamate-based inhibitor batimastat. The characteristic of the DDR1 sheddase to be blocked by batimastat suggests that it belongs to the membrane-bound matrix metalloproteinase or disintegrin and metalloproteinase family of proteases. PMID- 11943147 TI - Cracking the folding code. Why do some proteins adopt partially folded conformations, whereas other don't? AB - Many, but not all, globular proteins have been shown to have compact intermediate state(s) under equilibrium conditions in vitro, giving rise to the question: why do some proteins adopt partially folded conformations, whereas other do not? Here we show that charge to hydrophobicity ratio of a polypeptide chain may represent a key determinant in this respect, as proteins known to form equilibrium partially folded intermediates are specifically localized within a unique region of charge-hydrophobicity space. Thus, the competence of a protein to form equilibrium intermediate(s) may be determined by the bulk content of hydrophobic and charged amino acid residues rather than by the positioning of amino acids within the sequence. PMID- 11943148 TI - The actin-binding protein Filamin-A interacts with the metabotropic glutamate receptor type 7. AB - A yeast two-hybrid screen identified Filamin-A as a binding partner of the metabotropic glutamate receptor type 7b (mGluR7b) splice variant. In addition, Filamin-A interacted with mGluR4a, mGluR5a, mGluR5b, mGluR7a and mGluR8a. Domain mapping revealed that alternative splicing of mGluR4, mGluR7 and mGluR8 C-termini regulated the interaction. A conserved tyrosine within mGluR C-termini was identified to mediate the binding to Filamin-A. Protein interactions were verified in biochemical assays using recombinant and native proteins. Finally, co expression of Filamin-A and mGluR7 splice variants was shown in brain regions. These findings suggest that Filamin-A may physically link metabotropic glutamate receptors to the actin cytoskeleton. PMID- 11943149 TI - Mutations converting cyclodextrin glycosyltransferase from a transglycosylase into a starch hydrolase. AB - Cyclodextrin glycosyltransferase (CGTase) efficiently catalyzes transglycosylation of oligo-maltodextrins, although the enzyme also has a low hydrolytic activity. Its +2 substrate binding subsite, which contains the conserved Phe184 and Phe260 residues, has been shown to be important for this transglycosylation activity [Nakamura et al. (1994) Biochemistry 33, 9929-9936]. Here we show that the amino acid side chain at position 260 also controls the hydrolytic activity of CGTase. Three Phe260 mutants of Thermoanaerobacterium thermosulfurigenes CGTase were obtained with a higher hydrolytic activity than ever observed before for a CGTase. These Phe260 mutations even changed CGTase from a transglycosylase into a starch hydrolase. PMID- 11943151 TI - Radiation-induced apoptosis in MOLT-4 cells requires de novo protein synthesis independent of de novo RNA synthesis. AB - We investigated the effects of inhibition of de novo RNA and protein synthesis in ionizing radiation (IR)-induced apoptosis in the human T cell line MOLT-4. We observed that pretreatment with cycloheximide inhibited IR-induced apoptosis. However, pretreatment with actinomycin D did not inhibit apoptosis induced by IR. These results suggest that apoptosis induced by IR in MOLT-4 cells requires de novo protein synthesis but not de novo RNA synthesis. This finding suggests that the mRNA encoding the proapoptotic protein(s) is stabilized to facilitate translation independent of de novo gene transcription in response to IR. Our results also indicate that translation of the required proapoptotic protein(s) occurs upstream of mitochondrial depolarization and after 2 h post-IR. PMID- 11943150 TI - Involvement of the telomeric protein Pin2/TRF1 in the regulation of the mitotic spindle. AB - Pin2/TRF1 was independently identified as a telomeric DNA-binding protein (TRF1) that regulates telomere length, and as a protein (Pin2) that can bind the mitotic kinase NIMA and suppress its lethal phenotype. We have previously demonstrated that Pin2/TRF1 levels are cell cycle-regulated and its overexpression induces mitotic arrest and then apoptosis. This Pin2/TRF1 activity can be potentiated by microtubule-disrupting agents, but suppressed by phosphorylation of Pin2/TRF1 by ATM; this negative regulation is critical in mediating for many, but not all, ATM dependent phenotypes. Interestingly, Pin2/TRF1 specifically localizes to mitotic spindles in mitotic cells and affects the microtubule polymerization in vitro. These results suggest a role of Pin2/TRF1 in mitosis. However, nothing is known about whether Pin2/TRF1 affects the spindle function in mitotic progression. Here we characterized a new Pin2/TRF1-interacting protein, EB1, that was originally identified in our yeast two-hybrid screen. Pin2/TRF1 bound EB1 both in vitro and in vivo and they also co-localize at the mitotic spindle in cells. Furthermore, EB1 inhibits the ability of Pin2/TRF1 to promote microtubule polymerization in vitro. Given that EB1 is a microtubule plus end-binding protein, these results further confirm a specific interaction between Pin2/TRF1 and the mitotic spindle. More importantly, we have shown that inhibition of Pin2/TRF1 in ataxia telangiectasia cells is able to fully restore their mitotic spindle defect in response to microtubule disruption, demonstrating for the first time a functional involvement of Pin2/TRF1 in mitotic spindle regulation. PMID- 11943152 TI - Functional distinction of human EAG1 and EAG2 potassium channels. AB - Human ether a go-go potassium channel 2 (hEAG2) was cloned and its properties were compared with the previously characterized isoform hEAG1. In the Xenopus oocyte expression system the time course of activation was about four times slower and the voltage required for half-maximal subunit activation was about 10 mV greater for hEAG2 channels. However, its voltage dependence was smaller and, therefore, hEAG2 channels start to open at more negative voltages than hEAG1. Coexpression of both isoforms and kinetic analysis of the resulting currents indicated that they can form heteromeric channel complexes in which the slow activation phenotype of hEAG2 is dominant. Upon expression in mammalian cells, quinidine blocked hEAG1 channels (IC(50) 1.4 microM) more potently than hEAG2 channels (IC(50) 152 microM), thus providing a useful tool for the functional distinction between hEAG1 and hEAG2 potassium channels. PMID- 11943153 TI - Role of N-glycans and SDF-1alpha on the coassociation of CD4 with CXCR4 at the plasma membrane of monocytic cells and blood lymphocytes. AB - CXCR4 is a coreceptor, along with CD4, for human immunodeficiency virus type 1 (HIV-1). Trimolecular complexes between HIV-1 glycoprotein (gp)120, CD4 and CXCR4 constitute a prerequisite for HIV entry. We studied whether CD4 is associated with CXCR4 on CD4+ CXCR4+ cells. Using the conformation-dependent anti-CXCR4 mAb 12G5, CD4 was coimmunoprecipitated with CXCR4 from the membrane of U937 cells which support HIV-1(LAI) efficient infection, and from that of peripheral blood lymphocytes (PBL). CD4 association with CXCR4 increased upon PBL coculture for 5 days with autologous monocytes, decreased upon treatment of the cells or the CD4 CXCR4 complex with either N-glycanase or stromal cell derived factor-1alpha (SDF 1alpha) and was abolished by incubation of the cells with both, N-glycanase and SDF-1alpha. This indicates that glycans are partly involved in CD4 association with CXCR4 and may partly explain the inhibitory effect of SDF-1alpha on HIV infection. PMID- 11943154 TI - Membrane distal cytokine binding domain of LIFR interacts with soluble CNTFR in vitro. AB - Ciliary neurotrophic factor (CNTF) is a member of the gp130 family of cytokines. The functional receptor complex of CNTF is composed of the CNTF receptor alpha (CNTFR), gp130 and the leukemia inhibitory factor receptor (LIFR). Three regions on CNTF have been identified as binding sites for its receptors. The ligand receptor interactions are mediated through the cytokine binding domains (CBDs) and/or the immunoglobulin-like domains of the receptors. However, in the case of CNTF, the precise nature of the protein-protein contacts in the signaling complex has not yet been resolved. In this study, we provide the first demonstration that the membrane distal CBD (CBD1) of LIFR associates in vitro with soluble CNTFR in the absence of CNTF. Moreover, purified CBD1 partially blocks CNTF signaling, but not that of interleukin-6 or LIF, in human embryonal carcinoma cell line Ntera/D1 cells. These data raise the possibility that LIFR has the capability to form a ligand-free complex with CNTFR. PMID- 11943155 TI - CYP98A6 from Lithospermum erythrorhizon encodes 4-coumaroyl-4' hydroxyphenyllactic acid 3-hydroxylase involved in rosmarinic acid biosynthesis. AB - Rosmarinic acid is the dominant hydroxycinnamic acid ester accumulated in Boraginaceae and Lamiaceae plants. A cytochrome P450 cDNA was isolated by differential display from cultured cells of Lithospermum erythrorhizon, and the gene product was designated CYP98A6 based on the deduced amino acid sequence. After expression in yeast, the P450 was shown to catalyze the 3-hydroxylation of 4-coumaroyl-4'-hydroxyphenyllactic acid, one of the final two steps leading to rosmarinic acid. The expression level of CYP98A6 is dramatically increased by addition of yeast extract or methyl jasmonate to L. erythrorhizon cells, and its expression pattern reflected the elicitor-induced change in rosmarinic acid production, indicating that CYP98A6 plays an important role in regulation of rosmarinic acid biosynthesis. PMID- 11943156 TI - SufC hydrolyzes ATP and interacts with SufB from Thermotoga maritima. AB - Genetic experiments in bacteria have shown the suf operon is involved in iron homeostasis and the oxidative stress response. The sufB and sufC genes that always occur together in bacteria are also found in plants, and even the malaria parasite, associated with the plastid organelle. Although the suf operon is believed to encode an iron-dependent ABC-transporter there is no direct evidence. By immunolocalization we show here that SufB and SufC are associated with the membrane of Escherichia coli. We also present kinetic studies with a recombinant version of SufC from Thermotoga maritima that shows it is an ATPase and that it interacts with SufB in vitro. PMID- 11943157 TI - The advantages of cDNA microarray as an effective tool for identification of reproductive organ-specific genes in a model legume, Lotus japonicus. AB - To understand the molecular mechanisms intrinsic to reproductive organ development a cDNA microarray, fabricated from flower bud cDNA clones, was used to isolate genes, which are specifically expressed during the development of the anther and pistil in Lotus japonicus. Cluster analysis of the microarray data revealed 21 and 111 independent cDNA groups, which were specifically expressed in immature and mature anthers, respectively. RT-PCR was performed to provide a direct assessment of the accuracy and reproducibility of our approach. Confirmation of our results suggests that cDNA microarray technology is an effective tool for identification of novel reproductive organ-specific genes. PMID- 11943158 TI - Cell-cell signaling by direct contact increases cell proliferation via a PI3K dependent signal. AB - We report a novel mechanism of cellular growth control. Increasing the density of endothelial or smooth muscle cells in culture increased cell-cell contact and decreased cell spreading, leading to growth arrest. Using a new method to independently control cell-cell contact and cell spreading, we found that introducing cell-cell contact positively regulates proliferation, but that contact-mediated proliferation can be masked by changes in cell spreading: Round cells with many contacts proliferated less than spread cells with none. Physically blocking cell-cell contact or inhibiting PI3K signaling abrogated cell cell induced proliferation, but inhibiting diffusible paracrine signaling did not. Thus, direct cell-cell contact induces proliferation in these cells. PMID- 11943159 TI - Crystal structure of plant pectin methylesterase. AB - Pectin is a principal component in the primary cell wall of plants. During cell development, pectin is modified by pectin methylesterases to give different properties to the cell wall. This report describes the first crystal structure of a plant pectin methylesterase. The beta-helical structure embodies a central cleft, lined by several aromatic residues, that has been deduced to be suitable for pectin binding. The active site is found at the center of this cleft where Asp157 is suggested to act as the nucleophile, Asp136 as an acid/base and Gln113/Gln135 to form an anion hole to stabilize the transition state. PMID- 11943160 TI - Farnesylpyridinium, an analog of isoprenoid farnesol, induces apoptosis but suppresses apoptotic body formation in human promyelocytic leukemia cells. AB - 1-Farnesylpyridinium (FPy), an analog of isoprenoid farnesol, initially induced morphological changes similar to those of typical apoptosis in human leukemia HL 60 cells but FPy-treated cells were characterized by the absolute absence of final apoptotic events such as fragmentation into apoptotic bodies. FPy-induced cell death was considered to be apoptotic on the basis of the induction of DNA fragmentation and the protection against these events by the coaddition of a pan caspase inhibitor. The increase in the cytoplasmic cytochrome c level supported the possibility that FPy-treated cells should have the ability to complete the entire apoptotic process ending in cell fragmentation and apoptotic body formation. At concentrations too low to induce apoptosis, FPy could suppress the induction of apoptotic body formation in HL-60 cells by typical inducers of apoptosis such as actinomycin D or anisomycin. FPy exhibited a cytochalasin-like effect on spatial arrangement of actin filament independent of its apoptosis inducing activity. PMID- 11943161 TI - The putative effector-binding site of Leishmania mexicana pyruvate kinase studied by site-directed mutagenesis. AB - The activity of pyruvate kinase of Leishmania mexicana is allosterically regulated by fructose 2,6-bisphosphate (F-2,6-P(2)), contrary to the pyruvate kinases from other eukaryotes that are usually stimulated by fructose 1,6 bisphosphate (F-1,6-P(2)). Based on the comparison of the three-dimensional structure of Saccharomyces cerevisiae pyruvate kinase crystallized with F-1,6 P(2) present at the effector site (R-state) and the L. mexicana enzyme crystallized in the T-state, two residues (Lys453 and His480) were proposed to bind the 2-phospho group of the effector. This hypothesis was tested by site directed mutagenesis. The allosteric activation by F-2,6-P(2) appeared to be entirely abrogated in the mutated enzymes confirming our predictions. PMID- 11943162 TI - Bruton's tyrosine kinase regulates B cell antigen receptor-mediated JNK1 response through Rac1 and phospholipase C-gamma2 activation. AB - Bruton's tyrosine kinase (Btk) is essential for B cell development and B cell antigen receptor (BCR) function. Recent studies have shown that Btk plays an important role in BCR-mediated c-Jun NH(2)-terminal kinase (JNK) 1 activation; however, the mechanism by which Btk participates in the JNK1 response remains elusive. Here we show that the BCR-mediated Rac1 activation is significantly inhibited by loss of Btk, while this Rac1 activation is not affected by loss of phospholipase C-gamma2 (PLC-gamma2). Since PLC-gamma2 is also required for BCR mediated JNK1 response, our results suggest that Btk regulates Rac1 pathway as well as PLC-gamma2 pathway, both of which contribute to the BCR-mediated JNK1 response. PMID- 11943163 TI - Direct interaction of soluble human recombinant tau protein with Abeta 1-42 results in tau aggregation and hyperphosphorylation by tau protein kinase II. AB - We report here that aggregated beta-amyloid (Abeta) 1-42 promotes tau aggregation in vitro in a dose-dependent manner. When Abeta-mediated aggregated tau was used as a substrate for tau protein kinase II (TPK II), an 8-fold increase in the rate of TPK II-mediated tau phosphorylation was observed. The extent of TPK II dependent tau phosphorylation increased as a function of time and Abeta 1-42 concentration, and hyperphosphorylated tau was found to be decorated with an Alzheimer's disease-related phosphoepitope (P-Thr-231). In HEK 293 cells co expressing CT-100 amyloid precursor protein and tau, the release of Abeta 1-42 from these cells was impaired. Taken together, these in vitro results suggest that Abeta 1-42 promotes both tau aggregation and hyperphosphorylation. PMID- 11943164 TI - Nucleotide specificity of an archaeal group II chaperonin from Thermococcus strain KS-1 with reference to the ATP-dependent protein folding cycle. AB - The archaeal chaperonin-mediated folding of green fluorescent protein (GFP) was examined in the presence of various nucleotides. The recombinant alpha- and beta subunit homo-oligomers and natural chaperonin oligomer from Thermococcus strain KS-1 exhibited folding activity with not only ATP but also with CTP, GTP, or UTP. The ADP-bound form of both recombinant and natural chaperonin had the ability to capture non-native GFP, but could not refold it in the presence of CTP, GTP or UTP until ATP was supplied. The archaeal chaperonin thus utilized ATP, but could not use other nucleoside triphosphates in the cytoplasm where ADP was present. PMID- 11943165 TI - Recombinant glucocorticoid induced tumor necrosis factor receptor (rGITR) induces NOS in murine macrophage. AB - Glucocorticoid induced tumor necrosis factor receptor (GITR) is a new member of the tumor necrosis factor-nerve growth factor receptor superfamily of which the function has not been well studied. The extracellular domain of GITR was produced in Escherichia coli and purified as a single band of predicted M(r) of 18.0 kDa. GITR and GITR ligand were expressed constitutively on the surface of Raw 264.7 macrophage cell line and murine peritoneal macrophages. An extracellular domain of GITR can activate murine macrophages to express inducible nitric oxide synthase and to generate nitric oxide in a dose- and time-dependent manner. PMID- 11943166 TI - Kinetic mechanism and order of substrate binding for sn-glycerol-3-phosphate acyltransferase from squash (Cucurbita moschata). AB - sn-Glycerol-3-phosphate acyltransferase (G3PAT, EC 2.3.1.15), a component of glycerolipid biosynthesis, is an important enzyme in chilling sensitivity in plants. The three-dimensional structure of the enzyme from squash (Cucurbita moschata), without bound substrate, has been determined [Turnbull et al. (2001) Acta Crystallogr. D 57, 451-453; Turnbull et al. (2001) Structure 9, 347-353]. Here we report the kinetic mechanism of plastidial G3PAT from squash and the order of substrate binding using acyl-acyl carrier protein (acyl-ACP) substrates. The reaction proceeds via a compulsory-ordered ternary complex with acyl-ACP binding before glycerol-3-phosphate. We have also determined that the reaction will proceed with C(4:0)-CoA, C(6:0)-CoA and C(12:0)-ACP substrates, allowing a wider choice of acyl groups for future co-crystallisation studies. PMID- 11943167 TI - Regulation of fibronectin alternative splicing by a basement membrane-like extracellular matrix. AB - Hepatocytes are the source of plasma fibronectin (FN) which lacks the alternatively spliced EDI segment, distinctive of oncofetal FN. When hepatic or other epithelial cells are cultured on plastic, EDI inclusion is triggered. Here we report that EDI inclusion is inhibited when hepatic cells are cultured on a basement membrane-like extracellular matrix (ECM), demonstrating a new role for the ECM in the control of gene expression. The effect is duplicated by collagen IV and laminin but not by collagen I; is not observed with another alternatively spliced FN exon (EDII); and correlates with a decrease in cell proliferation, consistently with high EDI inclusion levels observed in many physiological and pathological proliferative processes. PMID- 11943168 TI - Expression of Fab fragment of catalytic antibody 6D9 in an Escherichia coli in vitro coupled transcription/translation system. AB - The heavy chain (Hc) and light chain (Lc) genes of the Fab fragment of a catalytic antibody 6D9 were simultaneously expressed in an Escherichia coli in vitro transcription/translation system without a reducing agent. The intermolecular disulfide bond between the Hc and Lc was found formed, suggesting a correct formation of the Fab fragment in the in vitro system. In enzyme-linked immunosorbent assay, the Fab fragment synthesized in vitro exhibited an antigen binding activity. Addition of reduced glutathione, oxidized glutathione, protein disulfide-isomerase and molecular chaperones, GroEL and GroES, increased the solubility and the antigen-binding activity of the Fab fragment greatly. The in vitro synthesized Fab was purified by means of a hexa-histidine tag attached to the C-terminus of the Hc. Catalytic assay of the purified Fab fragment showed that the His-tagged Fab fragment synthesized in vitro had a catalytic activity comparable to that produced in vivo. PMID- 11943169 TI - pH dependence of the hydrogen exchange in the SH3 domain of alpha-spectrin. AB - Using nuclear magnetic resonance we have measured the hydrogen exchange (HX) in the Src homology region 3 (SH3) domain of alpha-spectrin as a function of pH*. At very acidic pH* values the exchange of most residues appears to occur via global unfolding, although several residues show abnormally large Gibbs energies of exchange, suggesting the presence of some residual structure in the unfolded state. At higher pH* HX occurs mainly via local or partial unfoldings. We have been able to characterize the coupling between the electrostatic interactions in this domain and the conformational fluctuations occurring under native conditions by analyzing the dependence upon pH* of the Gibbs energy of exchange. The SH3 domain seems to be composed of a central core, which requires large structural disruptions to become exposed to the solvent, surrounded by smaller subdomains, which fluctuate independently. PMID- 11943170 TI - Novel nuclear-encoded proteins interacting with a plastid sigma factor, Sig1, in Arabidopsis thaliana. AB - Sigma factor binding proteins are involved in modifying the promoter preferences of the RNA polymerase in bacteria. We found the nuclear encoded protein (SibI) that is transported into chloroplasts and interacts specifically with the region 4 of Sig1 in Arabidopsis. SibI and its homologue, T3K9.5 are novel proteins, which are not homologous to any protein of known function. The expression of sibI was tissue specific, light dependent, and developmentally timed. We suggest the transcriptional regulation by sigma factor binding proteins to function in the plastids of higher plant. PMID- 11943171 TI - N(6)-Adenine DNA-methyltransferase in wheat seedlings. AB - The N(6)-adenine DNA-methyltransferase was isolated from the vacuolar vesicle fraction of wheat coleoptiles. In the presence of S-adenosyl-L-methionine the enzyme de novo methylates the first adenine residue in the TGATCA sequence in the single- or double-stranded DNA substrates but it prefers single-stranded structures. Wheat adenine DNA-methyltransferase (wadmtase) is a Mg(2+)- or Ca(2+) dependent enzyme with a maximum activity at pH 7.5-8.0. Wadmtase seems to be responsible for mitochondrial DNA modification that might be involved in the regulation of replication of mitochondria in plants. PMID- 11943172 TI - Thyroid hormone increases transcription of GA-binding protein/nuclear respiratory factor-2 alpha-subunit in rat liver. AB - Thyroid hormone (TH) regulates mitochondrial respiratory rate by activating coordinated transcription in the nucleus and mitochondria. Whereas TH activates transcription of mitochondrial genes directly, the activation of nuclear-encoded mitochondrial genes is probably executed by indirect unknown mechanisms. Nuclear respiratory factors (NRF)-1 and GA-binding protein (BP)/NRF-2 may function as transacting genes, but regulation of these genes by TH is not demonstrated. We show that TH administration to hypothyroid rats promptly increases GABP/NRF-2 alpha-subunit mRNA levels in the liver, without significant changes in beta, gamma subunits. In run-on and time-course experiments, the transcription rate and protein levels increased three-fold in response to TH, indicating GABP/NRF-2 transcriptional regulation. The results also support the notion that ATP synthase beta-subunit is regulated by TH through the indirect activation of GABP/NRF-2. PMID- 11943173 TI - Dual action of isoprenols from herbal medicines on both PPARgamma and PPARalpha in 3T3-L1 adipocytes and HepG2 hepatocytes. AB - Several herbal medicines improve hyperlipidemia, diabetes and cardiovascular diseases. However, the molecular mechanism underlying this improvement has not yet been clarified. In this study, we found that several isoprenols, common components of herbal plants, activate human peroxisome proliferator-activated receptors (PPARs) as determined using the novel GAL4 ligand-binding domain chimera assay system with coactivator coexpression. Farnesol and geranylgeraniol that are typical isoprenols in herbs and fruits activated not only PPARgamma but also PPARalpha as determined using the chimera assay system. These compounds also activated full-length human PPARgamma and PPARalpha in CV1 cells. Moreover, these isoprenols upregulated the expression of some lipid metabolic target genes of PPARgamma and PPARalpha in 3T3-L1 adipocytes and HepG2 hepatocytes, respectively. These results suggest that herbal medicines containing isoprenols with dual action on both PPARgamma and PPARalpha can be of interest for the amelioration of lipid metabolic disorders associated with diabetes. PMID- 11943174 TI - Mechanism of allosteric modulation of Escherichia coli carbamoyl phosphate synthetase probed by site-directed mutagenesis of ornithine site residues. AB - The role of residues of the ornithine activator site is probed by mutagenesis in Escherichia coli carbamoyl phosphate synthetase (CPS). Mutations E783A, E783L, E892A and E892L abolish ornithine binding, E783D and T1042V decrease 2-3 orders of magnitude and E892D decreased 10-fold apparent affinity for ornithine. None of the mutations inactivates CPS. E783 mutations hamper carbamate phosphorylation and increase K(+) and MgATP requirements, possibly by perturbing the K(+)-loop near the carbamate phosphorylation site. Mutation E892A activates the enzyme similarly to ornithine, possibly by altering the position of K891 at the opening of the tunnel that delivers the carbamate to its phosphorylation site. T1042V also influences modulation by IMP and UMP, supporting signal transmission from the nucleotide effector to the ornithine site mediated by a hydrogen bond network involving T1042. Ornithine activation of CPS may be mediated by K(+)-loop and tunnel gating changes. PMID- 11943175 TI - Structural characterization of a protein pheromone from a cold-adapted (Antarctic) single-cell eukaryote, the ciliate Euplotes nobilii. AB - Free-living species of ciliated Protozoa control their vegetative (mitotic) proliferation and mating (sexual) processes by diffusible, cell type-specific protein signals (pheromones). One of these molecules, designated En-2, was isolated from a species, Euplotes nobilii, living in the stably cold marine waters of Antarctica, and its complete amino acid sequence of 60 residues was determined by automated Edman degradation of the whole protein and peptides generated by trypsin digestion. The proposed sequence is: DIEDFYTSETCPYKNDSQLA(20)WDTCSGGTGNCGTVCCGQCF(40)SFPVSQSCAGMADSNDCPNA(60). The En 2 structure appears to be characterized by an adaptive insertion of a glycine rich motif potentially capable to confer more flexibility to a functionally critical region of the molecule. PMID- 11943176 TI - A rapid fluorometric assay for the proteolytic activity of SKI-1/S1P based on the surface glycoprotein of the hemorrhagic fever Lassa virus. AB - The subtilase subtilisin kexin isozyme-1 (SKI-1)/site 1 protease (S1P), has been implicated in the processing of Lassa virus glycoprotein C (GP-C) precursor into GP1 and GP2 that are responsible for viral fusion with the host cell membrane. Here, we studied in vitro the kinetics of this cleavage by hSKI-1 using an intramolecularly quenched fluorogenic (IQF) peptide, Q-GPC(251-263) [Abz-(251)Asp Ile-Tyr-Ile-Ser-Arg-Arg-Leu-Leu/Gly-Thr-Phe-Thr(263)-3-NitroTyr-Ala-CONH(2)], containing the identified site. The measured V(max (app))/K(m (app)) was compared to those for other IQF SKI-substrates. Q-GPC(251-263) is cleaved 10-fold more efficiently than the previously known best SKI-substrate, Q-hproSKI(134-142). This study confirmed the role of SKI-1 in GP-C processing and provides a novel, rapid and efficient enzymatic assay of SKI-1. PMID- 11943177 TI - High free energy of lipid/protein interaction in biological membranes. AB - The free energy of lipid/protein interaction in biological membranes is still unknown although extensive partitioning and modelling studies have revealed many partial energetic increments. Multiple site binding kinetics are now applied to four well-studied functional membrane proteins, and mean free energy values (+/ S.D.) of -4.23+/-0.49 kcal/mol for single lipid binding sites and of -89.7+/-35.4 kcal/mol for complete lipid substitution are obtained. These high free energy values point to an important bioenergetic role of lipid/protein interaction in membrane functions. PMID- 11943178 TI - Functional analysis of the Arabidopsis thaliana GCPE protein involved in plastid isoprenoid biosynthesis. AB - Plastid isoprenoids are synthesized via the 2-C-methyl-D-erythritol 4-phosphate pathway. A few years after its discovery, most of the Escherichia coli genes involved in the pathway have been identified, including gcpE. In this work, we have identified an Arabidopsis thaliana protein with homology to the product of this gene. The plant polypeptide, GCPE, contains two structural domains that are absent in the E. coli protein: an N-terminal extension and a central domain of 30 kDa. We demonstrate that the N-terminal region targets the Arabidopsis protein to chloroplasts in vivo, consistent with its role in plastid isoprenoid biosynthesis. Although the presence of the internal extra domain may have an effect on activity, the Arabidopsis mature GCPE was able to complement a gcpE defective E. coli strain, indicating the plant protein is a true functional homologue of the bacterial gcpE gene product. PMID- 11943179 TI - A conserved proline residue is present in the transmembrane-spanning domain of Tom7 and other tail-anchored protein subunits of the TOM translocase. AB - The TOM translocase consists of several integral membrane proteins organised around the channel forming protein Tom40. Here we show that one of these protein subunits, Tom7, is a tail-anchored protein. The carboxy-terminal 33 amino acids of Tom7 contain the information for targeting the protein to the mitochondrial outer membrane, and a conserved proline residue within the transmembrane segment is required for efficient targeting of Tom7 to the outer membrane. An equivalent proline residue is important in targeting each of the other three tail-anchored proteins that associate with Tom40 to form the core of the TOM translocase. PMID- 11943180 TI - The SUPERMAN protein is an active repressor whose carboxy-terminal repression domain is required for the development of normal flowers. AB - SUPERMAN was identified as a putative regulator of transcription that acts in floral development, but its function remains to be clarified. We demonstrate here that SUPERMAN is an active repressor whose repression domain is located in the carboxy-terminal region. Ectopic expression of SUPERMAN that lacked the repression domain resulted in a phenotype similar to that of superman mutants, demonstrating that the repression activity of SUPERMAN is essential for the development of normal flowers. Constitutive expression of SUPERMAN resulted in a severe dwarfism but did not affect cell size, indicating that SUPERMAN might regulate genes that are involved in cell division. PMID- 11943181 TI - Functional evidence for the identification of an Arabidopsis clathrin light chain polypeptide. AB - Clathrin light chains (CLCs) are regulatory subunits of clathrin triskelia. Based on homology searches in Arabidopsis thaliana data bases we have identified three putative CLC clones, and have focused on the one with the highest homology to mammalian CLC sequences. Analysis of its sequence has revealed coiled-coil structures within a region that corresponds to the clathrin heavy chain-binding site. In addition there is a stretch of acidic amino acids, which is required for the regulatory function of CLC in clathrin assembly. This putative plant CLC ortholog, expressed in bacteria as a glutathione-S-transferase- and myc-tagged fusion protein, was shown to bind to CLC-free recombinantly expressed mammalian clathrin hubs. In contrast, purified native mammalian triskelia with endogeneous CLC did not bind the recombinant putative plant CLC. Based on the conserved sequences between the three Arabidopsis candidates it appears that plants, unlike mammals, may have more than two CLCs. PMID- 11943182 TI - Involvement of phosphatidylinositol 3-kinase in nuclear translocation of protein kinase C zeta induced by C2-ceramide in rat hepatocytes. AB - In this study we report that protein kinase C zeta (PKC zeta), one of the atypical isoforms of the PKC family located predominantly in cytosol, is redistributed by C2-ceramide treatment in isolated hepatocytes. PKC zeta increased in membrane and nuclear fractions after 30 min of treatment with C2 ceramide in a dose- and time-dependent manner. The action of C2-ceramide was inhibited by wortmannin and LY 294002, indicating that C2-ceramide-induced PKC zeta increase in both nucleus and membrane fractions is mediated by phosphatidylinositol 3-kinase (PI3-kinase) activation. In addition, a significant translocation of PI3-kinase to the nucleus was observed after C2-ceramide treatment. PMID- 11943184 TI - The gift of Gab. AB - Gab proteins, including mammalian Gab1, Gab2, Gab3, Drosophila DOS and Caenorhabditis elegans Soc1, comprise a growing family of scaffolding/docking molecules involved in multiple signaling pathways mediated by receptor tyrosine kinases (RTKs) and non-RTK receptors. This paper reviews the structure/function relationships of Gab proteins and their biological roles during normal growth, differentiation and development programs. PMID- 11943185 TI - Proteasome inhibitors prevent cytochrome c release during apoptosis but not in excitotoxic death of cerebellar granule neurons. AB - In order to find out whether and how proteasomes participate in the processes leading cerebellar granule cells to death either in necrosis, due to glutamate neurotoxicity, or in apoptosis, due to K(+) shift, we measured the three proteasome activities by using specific fluorescent probes and investigated the effect of several proteasome inhibitors, including MG132, on the cytochrome c release taking place in the early phase of both apoptosis and necrosis. We show that differently from apoptosis, the early phase of necrosis does not require proteasome activation. Inhibition of proteasome activity can prevent cytochrome c release in cerebellar granule cells undergoing apoptosis, thus improving cell survival, but not necrosis. These findings show that proteasomes play an important role in the early phase of apoptosis but not that of necrosis, and that these two types of cell death differ from each other in their mechanism of cytochrome c release. PMID- 11943186 TI - Site-directed mutagenesis of the type II TGF-beta receptor indicates a ligand binding site distinct from that of the type II activin receptor. AB - Site-directed mutagenesis was used to map the ligand-binding surface of the type II transforming growth factor-beta receptor extracellular domain (TbetaRII-ECD). Two putative ligand-binding sites were probed, the first being a predicted hydrophobic patch, the second being the finger 1 surface loop. Nine residues were mutated in the context of full-length TbetaRII and the effect of these mutations on ligand-binding and receptor signaling was analyzed. Complementary information was obtained by examining 'natural' evolutionary TbetaRII mutations. Together, the results indicate that residues within the finger 1 region, but not the hydrophobic patch, of the TbetaRII-ECD are required for productive ligand binding. We conclude that, surprisingly, the ECDs of TbetaRII and type II activin receptor utilize distinct interacting surfaces for binding their respective ligands. PMID- 11943187 TI - Arabidopsis glutathione-dependent formaldehyde dehydrogenase is an S nitrosoglutathione reductase. AB - S-Nitrosoglutathione (GSNO), an adduct of nitric oxide (NO) with glutathione, is known as a biological NO reservoir. Heterologous expression in Escherichia coli of a cDNA encoding a glutathione-dependent formaldehyde dehydrogenase of Arabidopsis thaliana showed that the recombinant protein reduces GSNO. The identity of the cDNA was further confirmed by functional complementation of the hypersensitivity to GSNO of a yeast mutant with impaired GSNO metabolism. This is the first demonstration of a plant GSNO reductase, suggesting that plants possess the enzymatic pathway that modulates the bioactivity and toxicity of NO. PMID- 11943188 TI - Genome-wide studies on the nuclear PDR3-controlled response to mitochondrial dysfunction in yeast. AB - Gain-of-function mutations in the transcription factors Pdr1p and Pdr3p lead to the up-regulation of genes controlling plasma membrane properties. Pdr3p is involved in a retrograde response in which mitochondrial dysfunctions activate PDR5, a gene encoding an ABC membrane transporter. We carried out genome-wide analyses of the PDR3-controlled genes activated by the deletion of the mitochondrial DNA. We present evidence showing that PDR1 does not interfere with this PDR3 response. We also showed that the mitochondrially activated PDR3 response is highly sensitive to both yeast strain variations and carbon sources. These observations explain the apparent discrepancies in published studies and better describe the connections between the mitochondrial state and plasma membrane properties. PMID- 11943189 TI - Lipid domain formation and ligand-receptor distribution in lipid bilayer membranes investigated by atomic force microscopy. AB - A novel experimental technique, based on atomic force microscopy (AFM), is proposed to visualize the lateral organization of membrane systems in the nanometer range. The technique involves the use of a ligand-receptor pair, biotin avidin, which introduces a height variation on a solid-supported lipid bilayer membrane. This leads to a height amplification of the lateral membrane organization that is large enough to be clearly imaged by scanning AFM. The power of the technique is demonstrated for a binary dipalmitoylphosphocholine diarachidoylphosphocholine lipid mixture which is shown to exhibit a distinct lateral lipid domain formation. The new and simple ligand-receptor-based AFM approach opens up new ways to investigate lipid membrane microstructure in the nanometer range as well as the lateral distribution of ligand-lipid and receptor protein complexes in supported membrane systems. PMID- 11943190 TI - Analysis of gene expression in rose petals using expressed sequence tags. AB - Single-pass sequences were obtained from the 5'-ends of a total of 1794 rose petal cDNA clones. Cluster analysis identified 242 groups of sequences and 635 singletons indicating that the database represents a total of 877 genes. Putative functions could be assigned to 1151 of the transcripts. Expression analysis indicated that transcripts of several of the genes identified accumulated specifically in petals and stamens. The cDNA library and expressed sequence tag database described here represent a valuable resource for future research aimed at improving economically important rose characteristics such as flower form, longevity and scent. PMID- 11943191 TI - A rapid cytokinin response assay in Arabidopsis indicates a role for phospholipase D in cytokinin signalling. AB - Seedlings of Arabidopsis thaliana harboring a fusion of the cytokinin-responsive ARR5 gene promoter and the GUS reporter gene were used for a pharmacological approach to study cytokinin signal transduction. The assay was shown to be rapid, sensitive, dose-dependent and highly specific for cytokinins, both adenine and phenylurea derivatives. Numerous inhibitors of known signalling pathways were tested and some were shown to suppress reporter gene induction. Particularly, primary alcohols that specifically inhibit phospholipase D (PLD) partially prevented cytokinin-induced GUS activity and reduced the accumulation of ARR5 gene transcripts. This indicates a role for PLD early during cytokinin signalling. PMID- 11943192 TI - Identification of differentially expressed genes by cDNA-AFLP technique during heat stress in cowpea nodules. AB - Legume nodules formed by diazotrophic microorganisms are active sites for biological nitrogen fixation (BNF). In tropical regions, a significant part of N supply for soybean, peanut and bean crops is derived from BNF, which is nevertheless often limited by high temperature stress. In contrast, cowpea nodules are very resistant to high temperatures. To understand the molecular bases of thermotolerance during BNF under heat stress, we have used cDNA amplified fragment length polymorphism experiments to identify differentially expressed transcripts from cowpea nodules subjected to heat shock treatment. The expression profiles obtained showed approximately 600 bands, 55 up-regulated and nine corresponding to genes repressed by heat stress. Twenty transcript-derived fragments were isolated, cloned and sequenced. The Vigna unguiculata nodule and stress response transcripts present similarities to those that encode low molecular weight heat shock proteins, wound-induced proteins, disease resistance protein, and xylan endohydrolase isoenzyme, as well as different housekeeping genes. The differential expression of 15 genes was confirmed by using Northern blot or reverse Northern hybridization experiments. PMID- 11943193 TI - The interferon-inducible gene, Ifi204, acquires malignant transformation capability upon mutation at the Rb-binding sites. AB - p204 overexpression in retinoblastoma (Rb)-/- mouse embryo fibroblasts or transfection of p204 mutated at both Rb-binding sites confer growth advantages, resulting in a significantly higher number of foci in a cell focus assay. To investigate the possibility that mutated p204 acquires malignant transformation capability, NIH3T3 cells were stably transfected with the expression vector pRcRSV204 double-mutant (p204dm) harboring both the C-terminal deletion up to amino acid 568 and the point mutation from glutamic acid to lysine at position 427, and analyzed for markers typical of cell immortalization and transformation. We detected a greater abundance of cell colonies in soft agar with p204dm expressing cells than vector control cells. The p204dm-transfected cells also displayed two other characteristics associated with malignant transformation, i.e. growth under low-serum conditions and formation of tumors in athymic nude mice. Moreover, their telomerase activity was significantly higher than in the vector control cells. It would thus seem that p204, devoid of functional Rb binding motifs, can become oncogenic. PMID- 11943194 TI - The hepoxilin analog PBT-3 inhibits heparin-activated platelet aggregation evoked by ADP. AB - We have previously shown that PBT-3, a stable synthetic analog of hepoxilins, inhibits the aggregation of human platelets in vitro evoked by collagen through inhibition of thromboxane A(2) formation and action on the TP receptor. We now show that PBT-3 is capable of potently inhibiting the second phase of aggregation evoked by ADP in both washed human platelets and platelet-rich plasma (PRP), a phase associated with thromboxane formation. Aspirin blocks this second phase as well; so does the thromboxane receptor antagonist SQ 29,548. When ADP-evoked aggregation in PRP is activated by heparin through an enhancement of thromboxane formation, PBT-3, aspirin as well as SQ 29,548 block this activation through different mechanisms. These data confirm the inhibitory action of PBT-3 on aggregation of human platelets through inhibition of both thromboxane formation and blockade of thromboxane receptor action and suggest that this family of compounds may be useful in the treatment of thrombotic disorders in combination with heparin. PMID- 11943195 TI - Mutant huntingtin aggregates do not sensitize cells to apoptotic stressors. AB - It has been postulated that neuronal inclusions composed of mutant huntingtin may play a causative role in the pathogenesis of Huntington's disease. To study the putative role of aggregates in modulating apoptotic vulnerability, SH-SY5Y cell lines stably expressing truncated huntingtin with 18 (wild-type) (N63-18Q) or 82 (mutant) (N63-82Q) glutamine repeats were established. Aggregates were observed in approximately 13% of the N63-82Q cells; no aggregates were observed in the N63 18Q cells. In response to apoptotic stimuli such as staurosporine or hyperosmotic stress, caspase-3 activity was significantly greater in the N63-82Q cells compared to the N63-18Q cells. However, double immunostaining for huntingtin and active caspase-3 revealed that the presence of aggregates did not correlate with the presence of active caspase-3, indicating that aggregates do not contribute to the increase in apoptosis in the N63-82Q cells. PMID- 11943196 TI - Caspase-3 is not essential for DNA fragmentation in MCF-7 cells during apoptosis induced by the pyrrolo-1,5-benzoxazepine, PBOX-6. AB - Effector caspases-3, -6 and -7 are responsible for producing the morphological features associated with apoptosis, such as DNA fragmentation. The present study demonstrates that a member of a novel series of pyrrolo-1,5-benzoxazepines, PBOX 6, induces apoptosis in MCF-7 cells, which lack caspase-3. Apoptosis was accompanied by DNA fragmentation and the activation of caspase-7, but not caspases-3 and -6. Inhibition of caspase-7 activity reduced the extent of apoptosis induced, indicating that activation of caspase-7 is involved in the mechanism by which PBOX-6 induces apoptosis in MCF-7 cells. This study suggests that caspase-3 is not necessarily essential for DNA fragmentation and the morphological changes associated with apoptosis. PMID- 11943197 TI - Inhibition of mitochondrial and plastid activity of Plasmodium falciparum by minocycline. AB - We previously reported the superior effect of minocycline against drug-resistant Plasmodium falciparum in vitro. Here, we report that RT-PCR for falciparum parasites treated with minocycline revealed reduced levels of RNA transcripts of the mitochondrion-encoded genes such as the COI and Cyb genes, as well as the plastid-encoded RNA polymerase subunit (rpoB/C) gene. However, we detected no apparent effects of the antibiotic on the transcription of merozoite surface antigen and small subunit rRNA genes encoded by the nucleus. In addition, treatment with chloroquine and pyrimethamine showed no substantial reduction of any RT-PCR products. These findings suggest that tetracycline antibiotics selectively inhibit both mitochondrial and plastid activity. PMID- 11943199 TI - The chicken-egg scenario of protein folding revisited. AB - What is the first step in protein folding - hydrophobic collapse (compaction) or secondary structure formation? It is still not clear if the major driving force in protein folding is hydrogen bonding or hydrophobic interactions or both. We analyzed data on the conformational characteristics of 41 globular proteins in native and partially folded conformational states. Our analysis shows that a good correlation exists between relative decrease in hydrodynamic volume and increase in secondary structure content. No compact equilibrium intermediates lacking secondary structure, or highly ordered non-compact species, were found. This correlation provides experimental support for the hypothesis that hydrophobic collapse occurs simultaneously with formation of secondary structure in the early stages of the protein folding. PMID- 11943198 TI - Nitric oxide affects plant mitochondrial functionality in vivo. AB - In this report, we show that nitric oxide affects mitochondrial functionality in plant cells and reduces total cell respiration due to strong inhibition of the cytochrome pathway. The residual respiration depends on the alternative pathway and novel synthesis of alternative oxidase occurs. These modifications are associated with depolarisation of the mitochondrial membrane potential and release of cytochrome c from mitochondria, suggesting a conserved signalling pathway in plants and animals. This signal cascade is triggered at the mitochondrial level and induces about 20% of cell death. In order to achieve a higher level of cell death, the addition of H(2)O(2) is necessary. PMID- 11943200 TI - p21-activated protein kinase gamma-PAK in pituitary secretory granules phosphorylates prolactin. AB - p21-activated protein kinase gamma-PAK phosphorylates prolactin (PRL) in rat pituitary secretory granules on Ser-177 and on the equivalent site, Ser-179, in recombinant human PRL. This is shown by comparison of phosphopeptide maps with the human PRL mutant S179D. gamma-PAK is present in rat and bovine granules as identified by in-gel phosphorylation of histone H4, and by immunoblotting. Thus, phosphorylation of PRL by gamma-PAK in granules produces the PRL molecule that has been shown to antagonize the growth-promoting activity of unmodified PRL, and is consistent with the identified role of gamma-PAK in the induction and maintenance of cytostasis. PMID- 11943201 TI - Identification of the novel proteins Yip4p and Yip5p as Rab GTPase interacting factors. AB - The Rab GTPases are key regulators of membrane traffic. Yip1p is a membrane protein of unknown function that has been reported to interact with the Rabs Ypt1p and Ypt31p. In this study we identify Yif1p, and two unknown open reading frames, Ygl198p and Ygl161p, which we term Yip4p and Yip5p, as Yip1p-related sequences. We demonstrate that the Yip1p-related proteins possess several features: (i) they have a common overall domain topology, (ii) they are capable of biochemical interaction with a variety of Rab proteins in a manner dependent on C-terminal prenylation, and (iii) they share an ability to physically associate with other members of the YIP1 family. PMID- 11943202 TI - Accelerated alpha-synuclein fibrillation in crowded milieu. AB - Parkinson's disease is the second most common age-related neurodegenerative disease, resulting from loss of dopaminergic neurons in the substantia nigra. The aggregation and fibrillation of alpha-synuclein has been implicated as a causative factor in the disease, and the process of fibril formation has been intensively studied in vitro with dilute protein solutions. However, the intracellular environment of proteins is crowded with other macromolecules, whose concentration can reach 400 g/l. To address this discrepancy, the effect of molecular crowding on alpha-synuclein fibrillation has being studied. The addition of high concentrations of different polymers (proteins, polysaccharides and polyethylene glycols) dramatically accelerated alpha-synuclein fibrillation in vitro. The magnitude of the accelerating effect depended on the nature of the polymer, its length and concentration. Our results suggest that the major factor responsible for the accelerated fibrillation under crowded conditions is the excluded volume. PMID- 11943203 TI - Analysis of respiratory mutants reveals new aspects of the control of glycogen accumulation by the cyclin-dependent protein kinase Pho85p. AB - The PHO85 gene of Saccharomyces cerevisiae encodes a cyclin-dependent protein kinase that can interact with 10 different cyclins (Pcls). In conjunction with Pcl8p and Pcl10p, Pho85p phosphorylates and regulates glycogen synthase. Respiratory-deficient strains, such as coq3 mutants, have reduced glycogen stores and contain hyperphosphorylated and inactive glycogen synthase. We show here that pho85 coq3 mutants have dephosphorylated and active glycogen synthase yet do not maintain glycogen reserves. In contrast, deletion of PCL8 and PCL10 in the coq3 mutant background partially restores glycogen accumulation. This suggested the existence of inputs from Pho85p into glycogen storage, independent of Pcl8p and Pcl10p, and acting antagonistically. PMID- 11943204 TI - Leptin directly stimulates thermogenesis in skeletal muscle. AB - Using a method involving repeated oxygen uptake (MO(2)) determinations in skeletal muscle ex vivo, the addition of leptin was found to increase MO(2) in soleus muscles from lean mice. These effects were found to be inhibited by phosphatidylinositol 3-kinase inhibitors, absent in muscles from obese Lepr(db) mice which have the dysfunctional long form of leptin receptor, and blunted in muscles from diet-induced obese mice in the fed state but not during fasting. These findings indicate that leptin has direct thermogenic effects in skeletal muscle, and that these effects require both the long form of leptin receptors and phosphatidylinositol 3-kinase signalling. PMID- 11943205 TI - Hydroxylated human homotrimeric collagen I in Agrobacterium tumefaciens-mediated transient expression and in transgenic tobacco plant. AB - Potential contamination of animal-derived collagen with pathogens has led to the demand for safe recombinant sources of this complex molecule. In continuation of our previous work [Ruggiero et al. (2000) FEBS Lett. 469, 132-136], here we show that it is possible to produce recombinant hydroxylated homotrimeric collagen in tobacco plants that are co-transformed with a human type I collagen and a chimeric proline-4-hydroxylase (P4H). This is to our knowledge the first time that transient expression in tobacco was used to improve the quality of a recombinant protein produced in plants through co-expression with an animal cell derived modifying enzyme. We demonstrated the functionality of the new chimeric P4H and thus improved the thermal stability of recombinant collagen I from plants to 37 degrees C. PMID- 11943206 TI - Differential activation of nuclear factor-kappaB by tumour necrosis factor receptor subtypes. TNFR1 predominates whereas TNFR2 activates transcription poorly. AB - Tumour necrosis factor-alpha (TNF-alpha) signals though two receptors, TNFR1 and TNFR2. TNFR1 has a role in cytotoxicity, whereas TNFR2 regulates death responses or proliferation. TNF activates pro-inflammatory transcription factor nuclear factor-kappaB (NF-kappaB) by uncertain signalling mechanisms. Here we report the contribution of each TNFR towards the NF-kappaB activation processes. In human cells expressing endogenous or exogenous TNFR2, in addition to TNFR1, we found both TNFRs capable of activating NF-kappaB, as measured by IkappaBalpha (inhibitor of NF-kappaB) degradation, electrophoretic mobility shift assay and NF kappaB gene reporter assays. TNFR2 activation did not degrade IkappaBbeta. However, TNF-effects on NF-kappaB activation occurred predominantly through TNFR1, with TNFR2 activating the transcription factor poorly. PMID- 11943207 TI - Dephosphorylation of the two regulatory components of myosin phosphatase, MBS and CPI17. AB - Dephosphorylation of the two key regulatory factors of myosin light chain phosphatase (MLCP), CPI17 and MBS (myosin binding subunit) of MLCP was studied. While Thr38 phosphorylated CPI17 is quite susceptible to protein phosphatases, phosphorylated MBS was highly resistant to dephosphorylation. Type 2A, 2B and 2C protein phosphatases (PP2A, PP2B and PP2C), but not type 1 (PP1), dephosphorylated CPI17. The majority of the CPI17 phosphatase activity in smooth muscle was attributed to PP2A and PP2C. Phospholipids inhibited dephosphorylation of MBS and arachidonic acid (AA) inhibited PP2A activity against both MBS and CPI17, raising the possibility that AA favors the preservation of active MLCP. Consistently, while the phosphorylation of CPI17 was promptly decreased when the agonist was removed, the phosphorylation of MBS was unchanged in intact smooth muscle fiber. The results suggest that MBS phosphorylation mediated regulation of MLCP is not suitable for regulating rapid change in myosin phosphorylation. On the other hand, phosphorylated CPI17 is readily dephosphorylated thus likely to play a role in regulating fast phosphorylation-dephosphorylation cycle in cells. PMID- 11943208 TI - Involvement of NADPH in the cyclization reaction of carotenoid biosynthesis. AB - Cyclic carotenoids, e.g. beta-carotene, are formed by cyclization of an acyclic precursor, lycopene. The gene, crtY, which encodes lycopene beta-cyclase, has a partial sequence characteristic of a pyridine nucleotide binding domain, and NAD(P)H has been reported to be an absolute requirement for the cyclization reaction in vitro. By complementary incubations with lycopene as substrate and with (4R)-[4-(2)H]NADPH in (1)H(2)O or with unlabelled NADPH in (2)H(2)O in the presence of the purified enzyme, it has now been shown that the hydrogen atom introduced at C(2) in the cyclization comes from water and not from NADPH. The previously proposed mechanism involving the initiation of cyclization by H(+) attack at C(2) of the folded acyclic end group of the precursor is thus confirmed. No hydrogen is transferred from NADPH, which is therefore not involved directly in the cyclization reaction, but must play an indirect role, e.g. as an allosteric activator. PMID- 11943210 TI - Mild heat shock induces cyclin D1 synthesis through multiple Ras signal pathways. AB - Hyperthermia such as that occurring during fever may improve cell survival during infection, although its mechanism of action is largely unknown. Here we show that acute exposure to mild, but not severe, heat shock induces the synthesis of cyclin D1 that plays a critical role(s) in G1 progression of the cell cycle. This induction seemed to be regulated through multiple Ras signal pathways involving extracellular signal-regulated kinase, phosphatidylinositol 3-kinase, and Rac1/NADPH oxidase, all of which have well been documented to be responsible for growth factor-induced cyclin D1 expression. In a physiological sense, mild heat shock may regulate cell proliferation through inducing cyclin D1 along with growth factors. PMID- 11943209 TI - Autotaxin promotes motility via G protein-coupled phosphoinositide 3-kinase gamma in human melanoma cells. AB - Autotaxin (ATX), an exo-nucleotide pyrophosphatase and phosphodiesterase, stimulates tumor cell motility at sub-nanomolar levels and augments invasiveness and angiogenesis. We investigated the role of G protein-coupled phosphoinositide 3-kinase gamma (PI3Kgamma) in ATX-mediated tumor cell motility stimulation. Pretreatment of human melanoma cell line A2058 with wortmannin or LY294002 inhibited ATX-induced motility. ATX increased the PI3K activity in p110gamma, but not p85, immunoprecipitates. This effect was abrogated by PI3K inhibitors or inhibited by pertussis toxin. Furthermore, stimulation of tumor cell motility by ATX was inhibited by catalytically inactive form of PI3Kgamma, strongly indicating the crucial role of PI3Kgamma for ATX-mediated motility in human melanoma cells PMID- 11943211 TI - Artificial in vivo antigen presentation by the APCs and subsequent T-cell activation: a feasibility analysis. AB - Inappropriate antigen presentation by the antigen-presenting cells (APCs) is a cause of various diseases. One of the ways to combat these diseases is to immobilize the APCs near the infected tissue or a tissue which is susceptible to an antigen. The antigen is presented by the APCs present in the immobilized form on an implant and these upon binding to T(H)-cells result in triggering of a cascade of events as part of the natural immune response leading to the destruction of the antigen. This system has been modeled as a dialysis bag containing immobilized receptors inside the bag and the ligand diffusing out of the bag. The simulations show that by using the implant, the concentration of the ligand that has diffused into the tissue matrix can be substantially reduced and by suitably choosing the coupler size, the T(H)-cells can also effectively be activated. PMID- 11943212 TI - Phosphorylation of microtubule-associated protein tau by stress-activated protein kinases in intact cells. AB - Tau is a microtubule-associated protein that is abnormally hyperphosphorylated in the filamentous lesions that define a number of neurodegenerative diseases collectively referred to as tauopathies. We previously showed that stress activated protein (SAP) kinases phosphorylate tau protein at many of the hyperphosphorylated sites in vitro. Here we have developed a system to study the effects of five SAP kinases (SAPK1c/JNK1, SAPK2a/p38alpha, SAPK2b/p38beta, SAPK3/p38gamma and SAPK4/p38delta) on tau phosphorylation in intact cells. All kinases phosphorylated tau, albeit at different efficiencies. Tau was a good substrate for SAPK3/p38gamma and SAPK4/p38delta, a reasonable substrate for SAPK2b/p38beta and a relatively poor substrate for SAPK2a/p38alpha and SAPK1c/JNK1. These findings indicate that the aberrant activation of SAP kinases, especially SAPK3/p38gamma and SAPK4/p38delta, could play an important role in the abnormal hyperphosphorylation of tau that is an invariant feature of the tauopathies. PMID- 11943213 TI - Troponin I converts the skeletal muscle ryanodine receptor into a rectifying calcium release channel. AB - The goal of our present studies has been to find novel ryanodine receptor (RyR1) interacting polypeptides that modulate the channel activity from the luminal side of RyR1. Using K(+) as charge carrier for recording of single channel events here we demonstrate a very unexpected observation that troponin I substantially alters RyR's gating behavior, and that RyR1 in association with troponin I becomes a rectifying Ca(2+) release channel. Troponin I rapidly locks the RyR1 in a non conducting state only at a negative holding potential, and only when applied to the luminal side; switching to a positive holding potential results in the channel returning to its original activity, immediately. A hypothesis is proposed to account for how an intraluminally located, positively charged molecule might function as a RyR1 regulator under physiological conditions. PMID- 11943215 TI - NMR studies of the aggregation of glucagon-like peptide-1: formation of a symmetric helical dimer. AB - Nuclear magnetic resonance (NMR) spectroscopy reveals that higher-order aggregates of glucagon-like peptide-1-(7-36)-amide (GLP-1) in pure water at pH 2.5 are disrupted by 35% 2,2,2-trifluoroethanol (TFE), and form a stable and highly symmetric helical self-aggregate. NMR spectra show that the helical structure is identical to that formed by monomeric GLP-1 under the same experimental conditions [Chang et al., Magn. Reson. Chem. 37 (2001) 477-483; Protein Data Bank at RCSB code: 1D0R], while amide proton exchange rates reveal a dramatic increase of the stability of the helices of the self-aggregate. Pulsed field gradient NMR diffusion experiments show that the TFE-induced helical self aggregate is a dimer. The experimental data and model calculations indicate that the dimer is a parallel coiled coil, with a few hydrophobic residues on the surface that may cause aggregation in pure water. The results suggest that the coiled coil dimer is an intermediate state towards the formation of higher aggregates, e.g. fibrils. PMID- 11943214 TI - Leukotactin-1/CCL15-induced chemotaxis signaling through CCR1 in HOS cells. AB - Leukotactin-1 (Lkn-1)/CCL15 is a recently cloned CC-chemokine that binds to the CCR1 and CCR3. Although Lkn-1 has been known to function as a chemoattractant for neutrophils, monocytes and lymphocytes, its cellular mechanism remains unclear. To understand the mechanism of Lkn-1-induced chemotaxis signaling, we examined the chemotactic activities of human osteogenic sarcoma cells expressing CCR1 in response to Lkn-1 using inhibitors of signaling molecules. Inhibitors of G(i)/G(o) protein, phospholipase C (PLC) and protein kinase Cdelta (PKCdelta) inhibited the chemotactic activity of Lkn-1 indicating that Lkn-1-induced chemotaxis signal is transduced through G(i)/G(o) protein, PLC and PKCdelta. The activities of PLC and PKCdelta were also enhanced by Lkn-1 stimulation. Chemotactic activity of Lkn-1 was inhibited by the treatment of cycloheximide and actinomycin D suggesting that newly synthesized proteins are needed for chemotaxis. Nuclear factor-kappaB (NF-kappaB) inhibitor reduced chemotactic activity of Lkn-1. DNA binding activity of NF-kappaB was also enhanced by Lkn-1 stimulation. These results suggest that Lkn-1 transduces the signal through G(i)/G(o) protein, PLC, PKCdelta, NF-kappaB and newly synthesized proteins for chemotaxis. PMID- 11943216 TI - Synthesis of novel internal standards for the quantitative determination of plasma ceramide trihexoside in Fabry disease by tandem mass spectrometry. AB - The concentration of globotriaosylceramide (ceramide trihexoside (CTH)) in the plasma of patients with Fabry disease has been determined quantitatively by tandem mass spectrometry (MS) using novel internal standards, [D4]C-16 CTH and C 17 CTH, which were synthesised enzymically from lyso-CTH using the reverse reaction of sphingolipid ceramide N-deacylase. C-17 CTH was also synthesised chemically from lyso-CTH. This strategy has also been used to prepare standards for the quantitative determination by MS of other glycosphingolipids. PMID- 11943217 TI - Rat ENaC expressed in Xenopus laevis oocytes is activated by cAMP and blocked by Ni(2+). AB - We used oocytes of the South African clawed toad Xenopus laevis to express the three subunits of the epithelial Na(+) channel from rat distal colon (rENaC). We combined conventional dual-microelectrode voltage-clamp with continuous capacitance (C(m)) measurements and noise analysis to evaluate the effects of cAMP and Ni(2+) on rENaC. Control oocytes or rENaC-expressing oocytes exhibited no spontaneous fluctuations in current. However, in rENaC-expressing oocytes amiloride induced a marked plateau-shaped rise of the power density spectra. Recordings using four different concentrations of amiloride revealed that the blocker-channel interactions were of the first order. A cocktail of the membrane permeant cAMP analogue chlorophenylthio-cAMP and IBMX (cAMP cocktail) increased amiloride-sensitive current (I(ami)) and conductance (G(ami)). Furthermore, C(m) was also increased following cAMP application, indicating an increase in plasma membrane surface area. Noise analysis showed that cAMP increased the number of active channels in the oocyte membrane while single-channel current decreased. From these data we conclude that cAMP triggered exocytotic delivery of preformed rENaCs to the plasma membrane. Ni(2+) (2.5 mM) inhibited about 60% of the rENaC current and conductance while C(m) remained unaffected. Noise analysis revealed that this inhibition could be attributed to a decrease in the apparent channel density, while single-channel current did not change significantly. These observations argue for direct effects of Ni(2+) on channel activity rather than induction of endocytotic removal of active channels from the plasma membrane. PMID- 11943218 TI - Targeted cytosolic delivery of hydrogel nanoparticles into HepG2 cells through engineered Sendai viral envelopes. AB - Hydrogel nanoparticles of cross-linked polyvinylpyrrolidone (PVP-NP) (35-50 nm in diameter) containing fluoresceinated dextran (FITC-Dx) were encapsulated in reconstituted Sendai viral envelopes containing only the fusion (F) protein (F virosomes(1)). Incubation of these loaded F-virosomes with human hepatoblastoma cells (HepG2) in culture resulted in membrane-fusion-mediated delivery of NPs to the cell cytoplasm, as inferred from the ability of cells to internalize FITC-Dx loaded PVP-NP (PVP(f)-NP) in the presence of azide (an inhibitor of the endocytotic process). Introduction of PVP(f)-NP into the HepG2 cells was assured by selective accumulation of FITC fluorescence in the cytosolic compartment. The structural integrity of the internalized PVP(f)-NP was also confirmed by fluorescence microscopy and ultracentrifugation analysis. The potential usefulness of PVP-NP-mediated cytosolic release of water soluble drugs both in vitro and in vivo has been established for the first time. PMID- 11943219 TI - Self-assembly of ATP synthase subunit c rings. AB - Subunit c of the H(+) transporting ATP synthase is an essential part of its membrane domain that participates in transmembrane proton conduction. The annular architecture of the subunit c from different species has been previously reported. However, little is known about the type of interactions that affect the formation of c-rings in the ATPase complex. Here we report that subunit c over expressed in Escherichia coli and purified in non-ionic detergent solutions self assembles into annular structures in the absence of other subunits of the complex. The results suggest that the ability of subunit c to form rings is determined by its primary structure. PMID- 11943220 TI - Proceedings of the 8th International Coccidiosis Conference and the 2001 annual scientific meeting of the Australian Society for Parasitology. July 9-13, 2001. Cairns, Australia. PMID- 11943221 TI - Presidential address: rediscovering parasites using molecular tools--towards revising the taxonomy of Echinococcus, Giardia and Cryptosporidium. PMID- 11943222 TI - Recent advances in the search for new anti-coccidial drugs. AB - Coccidia provide a rich hunting ground for drug-designers, as there are significant biochemical differences between the parasites and their hosts. Recent years have brought the discovery of the plastid and its possible metabolic machinery, characterisation of acidocalcisomes, reports on the apparent absence from some coccidia of a typical mitochondrion, and the discovery of the mannitol cycle and shikimate pathway in the parasites. Moreover, modern technologies such as genomics and proteomics are bringing new insights into the biochemistry of coccidia and highlighting possible drug targets in abundance. A major issue for would-be drug discoverers is to decide upon the targets to prioritise. This review provides an update on recent findings on how coccidia differ biochemically from vertebrates. It includes discoveries within coccidian parasites themselves but also uses findings in Plasmodium to provide an overview of biochemical features that may be characteristics of many apicomplexan parasites and so potential targets for broad-spectrum drugs. PMID- 11943223 TI - Vaccinology for control of apicomplexan parasites: a simplified language of immune programming and its use in vaccine design. AB - Most mammalian immune systems and parasites have co-evolved over the millennia, interacting within a common environment and communicating through a common language. This language is comprised of copious dialects in which a variety of host innate and acquired immune pathways actively interact with a multitude of parasite-specific survival strategies. Nonetheless, a simplified language is likely present since the same basic molecular and cellular mechanisms are associated with resistance or susceptibility to parasite infection. Protective immunity against protozoa within the phylum Apicomplexa (e.g. Cryptosporidia, Eimeria, Neospora, Plasmodia and Toxoplasma) is generally CD4+ T cell-dependent and elicited along the IL-12/IFN-gamma/iNOS effector axis. This simplified language can be decoded in part by significant advances in understanding naive T cell activation, differentiation and generation of immunologic memory. Vaccine adjuvants and new immunisation strategies for generation of more potent immunity can also be viewed through this common language lens. The aim of this paper is to summarise recently published fundamental immunology studies, their relevance through examples in specific coccidian-host immune dialects, and how this simplified language can be used for the more rationale design of parasite vaccine control strategies. PMID- 11943224 TI - Cryptosporidium virulence determinants--are we there yet? AB - Exposure to Cryptosporidium parvum in healthy individuals results in transient infection that may be asymptomatic or can result in self-limited diarrhoea. In contrast, acquired immune deficiency syndrome patients with cryptosporidiosis can experience severe manifestations of disease. Volunteer studies have demonstrated that as few as 10 oocysts can cause infection in otherwise healthy adults and that isolates from geographically diverse regions differ in infectivity and, perhaps, virulence. Variability in isolate pathogenicity and infectivity has also been seen in bovine and murine models, respectively. Furthermore, isolate specific differences in protein composition and in host immunoreactivity have been observed. The molecular basis for differences in pathogenicity is not understood. Determining which factors are responsible for host selectivity and for the initiation, establishment, and perpetuation of infection with Cryptosporidium is key to rational drug design and vaccine development. To date, no specific virulence factors have been unequivocally shown to individually cause direct or indirect damage to host tissues nor have mutant strains been produced that could prove that particular deletions result in less virulent strains. Nevertheless, a number of candidate molecules have been identified by immunological and molecular methods. Here, we review the salient characteristics of some of these putative virulence determinants, including molecules that are involved in adhesion, protein degradation and the modulation of the host responses. PMID- 11943225 TI - Molecular prospecting for cryptic species of nematodes: mitochondrial DNA versus internal transcribed spacer. AB - DNA sequence divergence at internal transcribed spacer regions (ITS-1 and ITS-2) was compared with divergence at mitochondrial cox1 or nad4 loci in pairs of congeneric nematode species. Mitochondrial sequences accumulate substitutions much more quickly than internal transcribed spacer, the difference being most striking in the most closely related species pairs. Thus, mitochondrial DNA may be the best choice for applications in which one is using sequence data on small numbers of individuals to search for potential cryptic species. On the other hand, internal transcribed spacer remains an excellent tool for DNA diagnostics (quickly distinguishing between known species) owing to its lower level of intraspecific polymorphism. PMID- 11943227 TI - IL-4 increases Simian immunodeficiency virus replication despite enhanced SIV immune responses in infected rhesus macaques. AB - It is widely believed that a Th1 type CD4 response is critical for enhancement of CD8 immunity and for controlling HIV-1 infection. Th2 type responses, such as what might be seen in a chronic parasitic infection, would sacrifice cellular immunity and thus benefit the virus at the expense of the host. However, there has been little direct examination of the hypothesis in a primate model system. Accordingly, the simian immunodeficiency virus (SIV) infected rhesus macaque model was used to investigate the impact of immunisation with SIV expressing DNA constructs and co-injection with IL-4 on the SIV specific immunological responses, lymphocyte cell counts, as well as the impact on viral load. IL-4 is a Th2 type cytokine, which enhances antibody production and inhibits a CD4 Th1 phenotype. Rhesus macaques were infected with 10 AID50 of SIVmac239 and treated with 9-[2-(phosphonomethoxy)propyl]adenine (PMPA) 9 weeks post-infection. During PMPA treatment, animals were immunised with plasmids that expressed the SIV proteins, env, rev, gag and pol. In addition, they were immunised with a construct that encoded the gene for IL-4. IL-4 co-immunisation increased the neutralizing antibody titres in this group. Importantly, the viral loads in animals vaccinated with IL-4 expressing plasmid increased during the immunisation regimens despite the higher neutralizing antibody titres. In addition, neutralizing antibodies did not correlate with viral set point prior to PMPA treatment, however, there was a correlation between viral loads and antibody titres following the treatment with PMPA. Antibody titres decreased following the suppression of viral load. Importantly, vaccination in the absence of IL-4 protected CD4 levels without increasing viral load. The data support the hypothesis that inappropriate immune bias toward a Th2 pathway would ultimately enhance disease progression. PMID- 11943226 TI - Infection of organotypic slice cultures from rat central nervous tissue with Neospora caninum: an alternative approach to study host-parasite interactions. AB - Neospora caninum is an apicomplexan parasite which has emerged as an important cause of bovine abortion worldwide. Abortion is usually triggered by reactivation of dormant bradyzoites during pregnancy and subsequent congenital infection of the foetus, where the central nervous system appears to be most frequently affected. We here report on an organotypic tissue culture model for Neospora infection which can be used to study certain aspects of the cerebral phase of neosporosis within the context of a three-dimensionally organised neuronal network. Organotypic slice cultures of rat cortical tissue were infected with N. caninum tachyzoites, and the kinetics of parasite proliferation, as well as the proliferation-inhibitory effect of interferon-gamma (IFN-gamma), were monitored by either immunofluorescence, transmission electron microscopy, and a quantitative PCR-assay using the LightCycler instrument, respectively. In addition, the neuronal cytoskeletal elements, namely glial acidic protein filaments as well as actin microfilament bundles were shown to be largely colocalising with the pseudocyst periphery. This organotypic culture model for cerebral neosporosis provides a system, which is useful to study the proliferation, ultrastructural characteristics, development, and the interactions of N. caninum within the context of neuronal tissue, which at the same time can be modulated and influenced under controlled conditions, and will be useful in the future to gain more information on the cerebral phase of neosporosis. PMID- 11943228 TI - Molecular biology techniques in parasite ecology. AB - Molecular techniques are increasingly being used to study the ecology of a variety of organisms. These techniques represent important tools for the study of the systematics, population genetics, biogeography and ecology of parasites. Here, we review the techniques that have been employed to study the ecology and systematics of parasites (including bacteria and viruses). Particular emphasis is placed on the techniques of isoenzyme electrophoresis, in situ hybridisation and nucleic acid amplification to characterise parasite/microbial communities. The application of these techniques will be exemplified using ticks, bacterial endosymbionts and parasitic protozoa. PMID- 11943229 TI - Distribution and evolution of the Anopheles punctulatus group (Diptera: Culicidae) in Australia and Papua New Guinea. AB - The members of the Anopheles punctulatus group are major vectors of malaria and Bancroftian filariasis in the southwest Pacific region. The group is comprised of 12 cryptic species that require DNA-based tools for species identification. From 1984 to 1998 surveys were carried out in northern Australia, Papua New Guinea and on islands in the southwest Pacific to determine the distribution of the A. punctulatus group. The results of these surveys have now been completed and have generated distribution data from more than 1500 localities through this region. Within this region several climatic and geographical barriers were identified that restricted species distribution and gene flow between geographic populations. This information was further assessed in light of a molecular phylogeny derived from the ssrDNA (18S). Subsequently, hypotheses have been generated on the evolution and distribution of the group so that future field and laboratory studies may be approached more systematically. This study suggested that the ability for widespread dispersal was found to have appeared independently in species that show niche-specific habitat preference (Anopheles farauti s.s. and A. punctulatus) and conversely in species that showed diversity in their larval habitat (Anopheles farauti 2). Adaptation to the monsoonal climate of northern Australia and southwest Papua New Guinea was found to have appeared independently in A. farauti s.s., A. farauti 2 and Anopheles farauti 3. Shared or synapomorphic characters were identified as saltwater tolerance (A. farauti s.s. and Anopheles farauti 7) and elevational affinities above 1500 m (Anopheles farauti 5, Anopheles farauti 6 and A. farauti 2). PMID- 11943230 TI - Adapting immunity with subunit vaccines: case studies with group A Streptococcus and malaria. AB - Although vaccines have widely been regarded as the most cost-effective way to improve public health, for some organisms new technological advances in vaccine design and delivery, incurring additional developmental costs, will be essential. These organisms are typically those for which natural immunity is either slow to develop or does not develop at all. Clearly, such organisms have evolved strategies to evade immune responses and innovative approaches will be required to induce a type of immune response which is both different to that which develops naturally and is effective. This article describes some approaches to develop vaccines for two such organisms (malaria parasites and Streptococcus pyogenes (group A Streptococcus)) that are associated with widespread mortality and morbidity, mostly in the poorest countries of the world. At this stage, the challenges are primarily scientific, but if these hurdles are surmounted then the challenges will become financial ones--developing much needed vaccines for people least able to afford them. PMID- 11943231 TI - Companion animal parasitology: a clinical perspective. AB - In recent years there have been many changes to the ways that clinical veterinary science is conducted and nowhere is this more evident than in companion animal practice. Veterinarians working with pet dogs and cats are facing new challenges associated with the emergence and re-emergence of parasitic diseases. Some, such as Neospora caninum, have been recently recognised; others like Giardia and Cryptosporidium have been reported with increasing frequency, in part as a result of laboratory tests with improved sensitivity and specificity. In many regions, the emergence of parasitic diseases has been a consequence of pet travel and exotic diseases pose a unique diagnostic challenge for the veterinarian, as the index of suspicion for these conditions may be absent. The ranges of certain vector-borne diseases such as babesiosis, hepatozoonosis, ehrlichiosis, leishmaniasis and dirofilariasis are extending due to ecological and climatic changes and enhanced by animals with subclinical infection returning home from endemic areas. In companion animal practice, veterinarians have the additional responsibility of providing accurate information about the zoonotic transmission of parasite infections from pets, especially to those most vulnerable such as children, the elderly and the immunocompromised. Effective education is vital to allay public concerns and promote responsible pet ownership. PMID- 11943232 TI - The conceptual basis for a new classification of the coccidia. AB - At the joint meeting of the 8th International Coccidiosis Conference and the Annual Scientific Meeting of the Australian Society for Parasitology in Palm Cove, Australia, in July 2001, a Controversial Roundtable was held on 'New classification of coccidia'. The aim of this Roundtable was to stimulate and encourage discussion and debate on current classification schemes for the group of parasitic protozoa known as the eimeriid coccidia. In the past, such classifications have been based only on phenotypic characters such as morphology, ultrastructure, life cycles, and host specificity. However, over the past 10-15 years, molecular phylogenetic studies on taxa of the eimeriid coccidia have revealed that several of the families, subfamilies, and genera that have been erected based on non-molecular characters are paraphyletic. Therefore, this Roundtable was an important forum for initial discussions on how a new and more comprehensive classification of the eimeriid coccidia, which takes into consideration both phenotypic and molecular characters, can be devised. The stimulus came from invited speakers who gave introductions into selected areas of taxonomy and classification. Following these introductions, a more general discussion with the audience addressed potential steps that may be taken in future work. This review is the immediate outcome of the Roundtable. It describes advantages and disadvantages of the use of phenotypic or molecular characters as the base for taxonomic schemes for eimeriid coccidia. It gives specific examples for drawbacks of current classifications based only on phenotypic characters as well as potential pitfalls associated with the use of only molecular phylogenies. It addresses current controversies as well as rules of taxonomy and nomenclature relevant for the eimeriid coccidia. Finally, it recommends the establishment of an international group of scientists to meet on a regular basis, stimulate further discussions, and give direction on how the final goal, i.e. a proposal for a revised, and widely accepted, classification of the eimeriid coccidia, may be achieved. PMID- 11943233 TI - Sustainable coccidiosis control in poultry production: the role of live vaccines. AB - The development of new methods of administering coccidiosis vaccines has facilitated their use in the hatchery and thereby improved prospects for the economic vaccination of broilers. The acquisition of protective immunity to Eimeria species is boosted by further exposure to infection after vaccination. Factors that affect the reproductive efficiency of non-attenuated and attenuated vaccines are considered and the key role that oocyst production plays in establishing and maintaining uniform immunity in a flock of chickens is discussed. In addition to immunisation, a possible advantage to the application of certain vaccines is that their use could repopulate poultry houses with drug sensitive organisms. Theoretical rotation programmes in which the use of drugs is alternated with that of vaccines are described. Variability of the cross protective immune response between strains of the same species should be considered during vaccine development and subsequent use. The significance of less common species of Eimeria, not included in all vaccines, also needs to be assessed. An important consideration is the occurrence of pathogens other than Eimeria (such as the bacterium Clostridium) in flocks given coccidiosis vaccines and the methods by which they might be controlled. More research is required into the relationship between bacterial and viral infections of poultry and coccidiosis vaccination. Vaccines need to be developed that are simple to apply and cost effective for use in areas of the world where small-scale poultry production is commonplace. In the near future it is likely that more live vaccines based upon oocysts derived from attenuated strains of Eimeria will be developed but in the longer term vaccines will be based on the selective presentation to the host of specific molecules that can induce protective immunity. This achievement will require significant investment from the private and public sectors, and, if successful, will facilitate the sustainable control of coccidiosis in poultry production. PMID- 11943234 TI - Diagnosis and seroepidemiology of Neospora caninum-associated bovine abortion. AB - A round table was conducted at the VIIIth International Coccidiosis Conference on Neospora diagnosis with particular emphasis on strategies to diagnose bovine abortion. The strength and weakness of different assays for Neospora caninum infection and whether these methods have resulted in the overdiagnosis of neosporosis was discussed. It was evident that each diagnostic method, namely histology, immunohistochemistry, molecular detection and serological assays were, under certain circumstances, valuable in assessing the role N. caninum in abortion. Histological, immunohistochemical and molecular detection assays are of outstanding importance for the examination of tissues of aborted foetuses. While histology and immunohistochemistry allow direct assessment of pathomorphological changes caused by infection, molecular detection assays such as PCR are superior because of higher sensitivity and specificity in identifying N. caninum in foetal tissues. Serological tests, such as ELISA, are useful in determining whether an animal has been infected with N. caninum. Seroepidemiological approaches allow one to assess an abortion problem at a herd level and when used in conjunction with certain statistical methods are able to confirm a suspected N. caninum associated abortion. PMID- 11943235 TI - Resistance to antiparasitic drugs: the role of molecular diagnosis. AB - Chemotherapy is central to the control of many parasite infections of both medical and veterinary importance. However, control has been compromised by the emergence of drug resistance in several important parasite species. Such parasites cover a broad phylogenetic range and include protozoa, helminths and arthropods. In order to achieve effective parasite control in the future, the recognition and diagnosis of resistance will be crucial. This demand for early, accurate diagnosis of resistance to specific drugs in different parasite species can potentially be met by modern molecular techniques. This paper summarises the resistance status of a range of important parasites and reviews the available molecular techniques for resistance diagnosis. Opportunities for applying successes in some species to other species where resistance is less well understood are explored. The practical application of molecular techniques and the impact of the technology on improving parasite control are discussed. PMID- 11943236 TI - Autonomic and emotion regulation in bereavement and depression. AB - OBJECTIVES: Prior research suggests important differences between depression and the depressed feelings experienced in the context of bereavement, despite some overlap. Differences include an increase in restlessness, suggesting underlying physiological differences between the groups. METHOD: This study examined the level of depressive symptoms, heart rate (HR), and heart rate variability (HRV), and coping style of 10 bereaved, 10 depressed, and 10 control participants. RESULTS: Bereaved participants showed significantly higher HR than either depressed or control participants, while there were no such differences in HRV. Level of depression in the bereaved group correlated negatively with HRV. Additional analyses showed that the use of passive coping had a marginally significant negative correlation with HRV in bereaved subjects. CONCLUSION: The present data suggest that differences in HR and HRV could be associated with increased cardiovascular fatalities in bereaved individuals, known as the "broken heart phenomenon." These physiological differences have potential implications for both the mental and physical health of the bereaved. PMID- 11943237 TI - Conversion disorder in children and adolescents: a 4-year follow-up study. AB - OBJECTIVE: To assess the outcome of conversion disorder in children and adolescents and to identify factors affecting the prognosis. METHOD: Forty adolescents with conversion disorder were reevaluated 4 years after their initial interview. Changes in demographic and clinical data and the presence of any mood and anxiety disorders were recorded using the Structured Clinical Interview for DSM-IV Axis I Disorder (SCID-I). In addition, Beck Depression Inventory (BDI) and State-Trait Anxiety Inventory (STAI) were applied. RESULTS: Thirty-four patients (85%) had completely recovered from their conversion symptoms and two patients had improved (5%), whereas only four (10%) were unchanged. Fourteen (35%) patients received the diagnosis of mood and/or anxiety disorder. Favourable outcome was associated with early diagnosis (P=.04) and good premorbid adjustment (P=.01). CONCLUSION: Conversion disorder has a favourable outcome in children and adolescents. However, mood and/or anxiety disorders are encountered at a considerable rate in these patients even after recovery from conversion symptoms. Long clinical follow-up seems appropriate in children and adolescents with conversion disorder. PMID- 11943238 TI - Distress and hopelessness among parents of children with congenital heart disease, parents of children with other diseases, and parents of healthy children. AB - OBJECTIVE: We examined differences in distress (i.e., depression, anxiety, and somatisation) and hopelessness (e.g., suicide ideation) among parents of congenital heart disease (CHD) children (PCCHD, n=1092), parents of children with other diseases (PCOD, n=112), and parents of healthy children (PHC, n=293). In addition, we determined the proportion of parents in each group whose scores in distress and hopelessness, respectively, exceeded norms for psychiatric outpatients (POPN) and depressed people, and identified determinants of distress and hopelessness among all parents, and the PCCHD. METHOD: The parents completed a questionnaire about such areas as distress and hopelessness. The design was cross-sectional and data were collected during 20 consecutive days. RESULTS: PCCHD were generally at higher risk of distress and hopelessness. A significant number of parents, in particular PCCHD, reported levels of distress and hopelessness within/above POPN and depressed people, respectively. Mothers within all parent groups had higher levels of distress and hopelessness than fathers, with the highest levels among mothers of children with CHD compared to mothers in the other groups. Fathers of children with CHD were doing worse than fathers belonging to the other groups. There were no differences between PCOD and PHC. Variables such as employment status and financial situation explained more of the variation in distress and hopelessness among parents than the diseases of their children. CONCLUSION: We corroborated previous findings and provide new insights into the experiences of PCCHD that may be of importance when considering intervention. Further research concerning the parents, in particular PCCHD, at risk of developing psychosocial problems is needed. PMID- 11943239 TI - Factors predicting communication about the diagnosis of maternal breast cancer to children. AB - OBJECTIVE: To identify factors influencing mothers' communication with their children about their diagnosis and initial treatment. DESIGN: Cross-sectional cohort. SETTING: Two breast cancer treatment centres. SUBJECTS: Thirty-two women with Stage I or Stage II breast cancer with 56 school-aged children. MAIN OUTCOME MEASURES: Semistructured interview regarding timing and extent of communication with children from the time that a problem was first identified to initial treatment, and children's reactions. RESULTS: Children were most likely to be told about their mother's illness after the diagnosis had been confirmed by biopsy but a minority were told nothing until after surgery and some nothing at all. The information children received did not necessarily include mention of cancer. Predictors of communication were child age, with older children being told earlier and more information, and maternal education, with children of more highly educated mothers being told less. Child gender was not related to communication. CONCLUSION: Mothers who are being investigated for breast cancer are likely to benefit from the opportunity to consult a supportive professional about communication with their children during all stages of their diagnosis and treatment, but particularly early on in the investigation when they are more reluctant to initiate discussions with their children. At later points, those with children of varying ages may need specific advice on ways to discuss their illness in relation to children's understanding. PMID- 11943240 TI - Slow cortical potentials in migraine families are associated with psychosocial factors. AB - OBJECTIVE: The aim of the study was the determination of the relationships between neurophysiological and psychosocial factors within the pathogenesis of migraine. METHODS: The contingent negative variation (CNV), parent-child interactions and theirs relationship were investigated in 30 families with a migraine child and 20 healthy families. RESULTS: (1) None of the groups of children, independent of diagnosis, differed according to amplitude or habituation of the CNV. (2) Parents from migraine families exerted significantly more control over migraine children compared with interactions with healthy brothers/sisters. (3) The strong relation between CNV habituation/amplitude and abnormal pattern of parent-child interactions (especially overwhelming dominance and control) was found only for young migraineurs. This relation was not seen in healthy families or for healthy siblings of migraine children. CONCLUSION: This study provides significant evidence for a strong influence of family interactions on the development and maintenance of neurophysiological abnormalities in the migraineous headache. The role of psychosocial factors in the etiopathogenesis of migraine has to be investigated in further studies in more detail. PMID- 11943241 TI - Assessing the assessment measures for menstrual cycle symptoms: a guide for researchers and clinicians. AB - A review of measures of menstrual cycle symptoms is provided. This will enable researchers and clinicians to make the appropriate choice of method for their study requirements. In recent years, these measures have taken the form of retrospective questionnaires (rating severity of symptoms from memory) and prospective diaries (daily checklists of symptoms). Many of these draw on aspects of the well-known retrospective questionnaires, the Menstrual Distress Questionnaire and Premenstrual Assessment Form, in their development and validation. Each measure will be briefly described, followed by comments on its development, psychometric properties and finally an evaluation of its strengths and weaknesses. It concludes with an examination of the implications arising from the review, and some recommendations that menstrual cycle researchers and clinicians may wish to consider, as they decide upon the most appropriate measure for their needs. PMID- 11943242 TI - Sense of coherence as a mediator between hostility and health: seven-year prospective study on female employees. AB - OBJECTIVE: We proposed and tested a model in which low sense of coherence (SOC) was hypothesized to underlie the association between hostility and health problems. METHODS: Structural equation modeling was based on cross-lagged 7-year follow-up data, relating to five measurement points in 433 female municipal employees. RESULTS: The mediated model fitted well with the data. After adjustment for baseline characteristics, hostility was associated with increased risk of health problems, as indicated by records of sickness absences and poor self-rated health. Incorporating SOC into the model attenuated this association by 33-50%, depending on the indicator of health. The mediated effect of SOC was stronger than that of an alternative mediator, depressive symptoms. CONCLUSION: Low SOC may be a psychological background factor partially underlying the adverse effect of hostility on ill health. PMID- 11943243 TI - Responsiveness of measures in the effort-reward imbalance questionnaire to organizational changes: a validation study. AB - OBJECTIVE: To examine the responsiveness of measures adopted in the effort-reward imbalance (ERI) questionnaire to organizational changes. METHOD: For the employees who had been affected by restructuring of their company due to the economic hardship, two consecutive questionnaire surveys were conducted over a specific period. A total of 544 full-time employees responded to both surveys. Changes in four summary measures from the situation-specific model component and the person-specific component, and items/subscales that constitute the questionnaire were evaluated. RESULTS: The summary measures on psychological deterioration in the total study population. The deterioration was prevalent in those employees who had presumably experienced the effects of stressful organizational changes related to the restructuring, while improvement in the summary measures was observed for those employees who were promoted during the period. On the whole, the measures for the items of the situation-specific component and subscales for the personal component changed in the expected direction. With regard to ERI, potentially stronger effects of multiple organizational changes on employees were indicated. CONCLUSION: Measurements of ERI at work are valid in terms of responsiveness to organizational changes. PMID- 11943244 TI - Dissociated autonomic regulation during stress and physical complaints. AB - OBJECTIVE: In order to evaluate the relevance of individual differences in the regulation of autonomic subsystems to psychosomatic processes, reactivity of the autonomic nervous system to an emotional stressor was related to the occurrence of physical complaints. In contrast to most other studies, the measures were related to the degree of actually experienced stress. METHODS: In a large sample (n=111), electrodermal activity (EDA) and heart rate variability (HRV) were obtained at rest and during emotional stress. Physical complaints were assessed by a standardized symptom checklist providing subscales for different physiological systems. RESULTS: Subjects who were actually stressed by the stress induction procedure but showed only weak reactivity of EDA, and subjects who were only slightly stressed but showed a marked EDA response reported a high amount of gastrointestinal complaints. A greater decrease of high-frequency HRV from the rest to the stress condition was associated with reports of relatively more cardiovascular complaints. CONCLUSION: Findings indicate that the link between regulation of EDA and gastrointestinal functions has to be localized in higher brain structures and support the hypothesis that minor autonomic dysregulation may represent one pathway linking negative affect and the development of physical disorders. PMID- 11943245 TI - Psychiatric health in a sexually transmitted infections clinic: effect on reattendance. AB - BACKGROUND AND OBJECTIVES: Sexually transmitted infections (STIs) remain a public health priority, but associated psychological morbidity has recently been ignored. One aspect of STI control is test of cure and further treatment at follow-up clinic appointments. We chose reattendance rates as a measure of compliance and assessed whether reattendance was related to psychological morbidity. METHODS: 938 STI clinic patients were offered the Hospital Anxiety and Depression Scale (HAD) and staff rated each patient's psychological health. Predictors of attendance, caseness and staff recognition were assessed by logistic regression. RESULTS: 401/774 [51.9%; 95% confidence interval (CI): 48.3 55.4%] patients, who completed the HAD, scored above threshold. Staff rated 151/743 (20.4%) patients as having "psychological problems." HAD caseness was not associated with attending arranged follow-up [adjusted odds ratio (adj. OR): 0.83 (0.49-2.05)]. Patients rated with psychological problems by staff were more likely to attend [adj. OR: 1.91 (1.02-3.60)]. CONCLUSIONS: Half of our sample had significant anxiety and staff should be more aware of this suffering. Our work suggests that such awareness might improve subsequent attendance. PMID- 11943250 TI - A health scare in the mass media. PMID- 11943251 TI - Oral contraceptives, parity, and cervical cancer. PMID- 11943252 TI - Iatrogenic legacy from diethylstilbestrol exposure. PMID- 11943253 TI - Improving the detection of childhood visual problems and eye disorders. PMID- 11943254 TI - Telling the truth about sex. PMID- 11943255 TI - Effect of oral contraceptives on risk of cervical cancer in women with human papillomavirus infection: the IARC multicentric case-control study. AB - BACKGROUND: Use of oral contraceptives could increase risk of cervical cancer; however the effect of human papillomavirus (HPV), the main cause of cervical cancer, is not usually taken into account. We aimed to assess how use of oral contraceptives affected risk of cervical cancer in women who tested positive for HPV DNA. METHODS: We pooled data from eight case-control studies of patients with histologically confirmed invasive cervical carcinoma (ICC) and from two studies of patients with carcinoma in situ (ISC). Information about use of oral contraceptives was obtained from personal interviews. Effects were estimated as odds ratios, with logistic-regression models adjusted for possible confounders. FINDINGS: 1465 of 1561 (94%) patients with ICC, 211 of 292 (72%) with ISC, and 255 of 1916 (13%) controls were positive for HPV DNA. Compared with never-users, patients who had used oral contraceptives for fewer than 5 years did not have increased risk of cervical cancer (odds ratio 0.73; 95% CI 0.52-1.03). The odds ratio for use of oral contraceptives was 2.82 (95% CI 1.46-5.42) for 5-9 years, and 4.03 (2.09-8.02) for use for 10 years or longer, and these risks did not vary by time since first or last use. INTERPRETATION: Long-term use of oral contraceptives could be a cofactor that increases risk of cervical carcinoma by up to four-fold in women who are positive for cervical HPV DNA. In the absence of worldwide information about HPV status, extra effort should be made to include long-term users of oral contraceptives in cervical screening programmes. PMID- 11943256 TI - Role of parity and human papillomavirus in cervical cancer: the IARC multicentric case-control study. AB - BACKGROUND: High parity has long been suspected of being associated with an increased risk of cervical cancer, but previous analyses of this association have not taken the strong effect of human papillomavirus (HPV) into account. To assess the role of reproductive factors in the progression from HPV infection to cancer, we did a pooled analysis including only HPV-positive women. METHODS: We pooled data from eight case-control studies on invasive cervical carcinoma (ICC) and two on in-situ carcinoma (ISC) from four continents. 1465 patients with squamous-cell ICCs, 211 with ISCs, 124 with adenocarcinomas or adenosquamous ICCs, and 255 control women, all positive for HPV DNA by PCR-based assays, were analysed. We calculated pooled odds ratios by means of unconditional multiple logistic regression models, and adjusted them for sexual and non-sexual confounding factors. The 95% CI were estimated by treating the odds ratio as floating absolute risk. FINDINGS: We found a direct association between the number of full term pregnancies and squamous-cell cancer risk: the odds ratio for seven full term pregnancies or more was 3.8 (95% CI 2.7-5.5) compared with nulliparous women, and 2.3 (1.6-3.2) compared with women who had one or two full-term pregnancies. There was no significant association between risk of adenocarcinoma or adenosquamous carcinoma and number of full-term pregnancies. INTERPRETATION: High parity increases the risk of squamous-cell carcinoma of the cervix among HPV positive women. A general decline in parity might therefore partly explain the reduction in cervical cancer recently seen in most countries. PMID- 11943257 TI - Hypospadias in sons of women exposed to diethylstilbestrol in utero: a cohort study. AB - BACKGROUND: Transgenerational effects of diethylstilbestrol (DES) have been reported in animals, but effects in human beings are unknown. Alerted by two case reports, we aimed to establish the risk of hypospadias in the sons of women who were exposed to DES in utero. METHODS: We did a cohort study of all sons of a Dutch cohort of 16284 women with a diagnosis of fertility problems. We used a mailed questionnaire assessing late effects of fertility treatment to identify boys with hypospadias. We compared the prevalence rate of hypospadias between boys with and without maternal DES exposure in utero. FINDINGS: 16284 mothers (response rate 67%) reported 8934 sons. The mothers of 205 boys reported DES exposure in utero. Four of these children were reported to have hypospadias. In the remaining 8729 children, only eight cases of hypospadias were reported (prevalence ratio 21.3 [95% CI 6.5-70.1]). All cases of hypospadias were medically confirmed. Maternal age or fertility treatment did not affect the risk of hypospadias. Children conceived after assisted reproductive techniques such as in-vitro fertilisation were not at increased risk of hypospadias compared with children conceived naturally (1.8, 0.6-5.7). INTERPRETATION: Our findings suggest an increased risk of hypospadias in the sons of women exposed to DES in utero. Although the absolute risk of this anomaly is small, this transgenerational effect of DES warrants additional studies. PMID- 11943258 TI - Trichomegaly following treatment with interferon alpha-2b. PMID- 11943259 TI - Effect of vitamins C and E on progression of transplant-associated arteriosclerosis: a randomised trial. AB - BACKGROUND: Cardiac transplantation is associated with oxidant stress, which may contribute to the development of accelerated coronary arteriosclerosis. We postulated that treatment with antioxidant vitamins C and E would retard the progression of transplant-associated arteriosclerosis. METHODS: In a double-blind prospective study, 40 patients (0-2 years after cardiac transplantation) were randomly assigned vitamin C 500 mg plus vitamin E 400 IU, each twice daily (n=19), or placebo (n=21) for 1 year. The primary endpoint was the change in average intimal index (plaque area divided by vessel area) measured by intravascular ultrasonography (IVUS). Coronary endothelium-dependent vasoreactivity was assessed with intracoronary acetylcholine infusions. IVUS, coronary vasoreactivity, and vitamin C and E plasma concentrations were assessed at baseline and at 1 year follow-up. All patients received pravastatin. Analyses were by intention to treat. FINDINGS: Vitamin C and E concentrations increased in the vitamin group (vitamin C 43 [SD 21] to 103 [43] mmol/L; vitamin E 24 [14] to 65 [27] mmol/L) but did not change in the placebo group (vitamin C 45 [15] vs 43 [16] mmol/L; vitamin E 27 [14] vs 27 [9] mmol/L; p<0.0001 for difference between groups). During 1 year of treatment, the intimal index increased in the placebo group by 8% (SE 2) but did not change significantly in the treatment group (0.8% [1]; p=0.008). Coronary endothelial function remained stable in both groups. INTERPRETATION: Supplementation with antioxidant vitamins C and E retards the early progression of transplant-associated coronary arteriosclerosis. PMID- 11943260 TI - A loss-of-function polymorphic mutation in the cytolytic P2X7 receptor gene and chronic lymphocytic leukaemia: a molecular study. AB - BACKGROUND: Chronic lymphocytic leukaemia (CLL) has a familial incidence nearly three times higher than expected for the general population and one predisposing factor might be an inherited failure of mechanisms involved in apoptosis of lymphocytes. Our aim was to ascertain whether or not a defect in a proapoptotic pathway, caused by a single nucleotide polymorphism that results in loss-of function of P2X7 in healthy individuals, was present in leukaemic B lymphocytes of patients with CLL. METHODS: We extracted genomic DNA from the peripheral blood leucocytes of 36 unrelated individuals with CLL, four individuals with familial CLL, and 46 age-matched controls. We sequenced a PCR product to detect mutations in exon 13 of P2X7. In most patients with CLL, we measured expression and function of the P2X7 receptor by flow cytometry in B lymphocytes and T lymphocytes. FINDINGS: The prevalence of the polymorphic mutation and the frequency of the mutant allele were three-fold greater in individuals with CLL than in white, elderly controls. Individuals homozygous for the polymorphic allele had no P2X7 receptor function and heterozygotes had half the mean function of that seen in individuals homozygous for the wildtype allele; amounts of ATP induced apoptosis varied accordingly. In two families, in which we studied a father-son pair and a sister-sister pair with CLL, loss of P2X7 function arose because of inheritance of one or two 1513A-->C alleles for P2X7. INTERPRETATION: Activation of the P2X7 receptor leads to apoptosis of lymphocytes in individuals with CLL, and reduced function of this receptor has an anti-apoptotic effect, resulting in an increase in B-cell numbers. Thus, inheritance of a loss-of function polymorphic mutation at position 1513 in the P2X7 gene could contribute to the pathogenesis of CLL. PMID- 11943261 TI - Over-the-counter medication and its effects. PMID- 11943262 TI - Genetic variations in HLA-B region and hypersensitivity reactions to abacavir. AB - Hypersensitivity to abacavir affects about 4% of patients who receive the drug for HIV-1 infection. We did a retrospective, case-control study to identify multiple markers in the vicinity of HLA-B associated with hypersensitivity reactions. HLA-B57 was present in 39 (46%) of 84 patients versus four (4%) of 113 controls (p<0 small middle dot0001). However, because of low numbers of women and other ethnic groups enrolled, these findings relate largely to white men. The lower sensitivity of HLA-B57 for predicting hypersensitivity to abacavir identified in this study compared with a previous report highlights that predictive values for markers will vary across populations. Clinical monitoring and management of hypersensitivity reactions among patients receiving abacavir must remain unchanged. PMID- 11943263 TI - Tamoxifen for breast cancer among hysterectomised women. AB - Tamoxifen is a candidate drug for prevention of breast cancer, although findings from trials have not been consistent. In our extended follow-up (median 81.2 months, IQR 66.0-87.2) of the Italian Tamoxifen Trial, this drug did not significantly protect against breast cancer in women at usual or slightly reduced risk of the disease (p=0.215). Use of hormone replacement therapy increased risk of breast cancer, and users of such treatment who were randomly allocated to tamoxifen had a rate of breast cancer that was close to that of never-users. So far, no woman has died from breast cancer in this study. Decisions about introduction of tamoxifen to reduce risk of breast cancer remain important and open questions. PMID- 11943265 TI - HMG CoA reductase inhibitor (statin) and aortic valve calcium. AB - There is no known pharmacological therapy for calcific aortic valvular sclerosis or stenosis. Because leaflet calcification occurs in areas of lipoprotein deposition, we hypothesised that 3-hydroxy-3-methylglutaryl-coenzyme A (HMG COA) reductase inhibitors (statins) might slow aortic valve calcium (AVC) accumulation. We retrospectively identified 65 patients who had undergone two electron-beam computed tomography scans at a mean (SD) interval of 2.5 (1.6) years. 28 (43%) patients were receiving statins. Patients who were treated with statins had a 62-63% lower median rate of AVC accumulation (p=0.006) and 44-49% fewer statin patients had definite AVC progression (p=0.043). These findings suggest that statins may decrease AVC accumulation. PMID- 11943264 TI - Leishmania in discarded syringes from intravenous drug users. AB - Needle sharing by intravenous drug users (IVDUs) has been proposed as providing an alternative, artificial, and anthroponotic cycle for leishmania transmission. We looked for parasites in syringes discarded by IVDUs using two different PCR techniques. Leishmania spp were detected in 65 (52%) of 125 syringes collected in southern Madrid, Spain, in 1998, and in 52 (34%) of 154 collected in southwestern Madrid in 2000-01. We found shared restriction fragment length polymorphisms in 12 of 65 positive samples tested, suggesting that syringe sharing can indeed promote the spread of leishmania clones among IVDUs. PMID- 11943266 TI - Arsenic contamination affects millions in Bangladesh. PMID- 11943268 TI - Aspirin resistance may increase death risk in some patients with heart disease. PMID- 11943270 TI - US report draws attention to preventable vision problems. PMID- 11943271 TI - Utter isolation in a cold climate: the Antarctic challenge. PMID- 11943272 TI - Public health experts concerned about "hikikomori". PMID- 11943273 TI - Canada's supreme court rejects broadcaster's appeal on doctors' libel verdict. PMID- 11943274 TI - South African court again tells government to increase access to AIDS drug. PMID- 11943275 TI - Ready for the next influenza pandemic? PMID- 11943276 TI - Health and environmental experts worried by forthcoming US data-quality act. PMID- 11943279 TI - Drug firms' representatives express concern over WHO's AIDS drug list. PMID- 11943280 TI - Antifungal drug resistance of pathogenic fungi. AB - Pathogenic fungi have many complex mechanisms of resistance to antifungal drugs. Information about the clinical, cellular, and molecular factors contributing to antifungal-drug resistance continues to accumulate. We critically review the diagnosis, epidemiology, and mechanisms of antifungal drug resistance of pathogenic fungi. Better understanding of this resistance should assist in developing better detection strategies for preventing and treating refractory mycoses in the future. PMID- 11943281 TI - Partial splenectomy: uses of error. PMID- 11943282 TI - Population, environment, disease, and survival: past patterns, uncertain futures. AB - Societies are exploring what sustainable development means for development choices. Increasingly, we recognise that human population health is not just an input to socioeconomic development, but is an essential outcome, and, over time, a marker of sustainability. There has been recent attention to how stocks of social and human capital precondition gains in population health. However, recognition of how environmental change can limit health and survival has been slower. Over many millennia, disease and longevity profiles in populations have reflected changes in environmental conditions and, often, excedances of carrying capacity. Today, population growth and the aggregated pressures of consumption and emissions are beginning to impair various global environmental systems. The research tasks in detecting, attributing, and projecting the resultant health effects are complex. Have recent health gains, in part, depended on depleting natural environmental capital? Population health sciences have a crucial contribution to make to the sustainability project. PMID- 11943283 TI - Evidence-based cellular pathology. PMID- 11943284 TI - Triptan medications to treat acute migraine. PMID- 11943285 TI - Triptan medications to treat acute migraine. PMID- 11943286 TI - Triptan medications to treat acute migraine. PMID- 11943287 TI - Triptan medications to treat acute migraine. PMID- 11943288 TI - Triptan medications to treat acute migraine. PMID- 11943290 TI - Cardioprotection of trimetazidine and anthracycline-induced acute cardiotoxic effects. PMID- 11943291 TI - Central venous pressure monitoring during pulmonary embolism. PMID- 11943292 TI - Study of genes and environmental factors in complex diseases. PMID- 11943293 TI - Study of genes and environmental factors in complex diseases. PMID- 11943294 TI - Study of genes and environmental factors in complex diseases. PMID- 11943295 TI - Study of genes and environmental factors in complex diseases. PMID- 11943297 TI - Vasopeptidase inhibitors: a bradykinin link. PMID- 11943298 TI - Leflunomide in treatment of rheumatoid arthritis. PMID- 11943300 TI - Parental attitudes towards passive smoking in Japan. PMID- 11943301 TI - Advanced trauma life support courses. PMID- 11943302 TI - Adaptation of bone to mechanical loads. PMID- 11943303 TI - Position of cardiac monitor and defibrillator. PMID- 11943307 TI - From the medical museum. The Lunatick's profits. PMID- 11943313 TI - The emerging role of avian cytokines as immunotherapeutics and vaccine adjuvants. AB - The use of antibiotic feed additives and chemical antimicrobials in food production animals is a double-edged sword. On one hand, it helps to prevent the outbreak of disease and promotes the growth of animals, but on the other hand, concerns are mounting over the emergence of antibiotic-resistant bacteria. As a consequence, some countries have already banned the use of in-feed antibiotics which has resulted in meat producers urgently seeking environmentally friendly alternative methods to control disease. Cytokines are proteins that control the type and extent of an immune response following infection or vaccination. They therefore represent excellent naturally occurring therapeutics. The use of cytokines in poultry has become more feasible with the discovery of a number of avian cytokine genes. Since the immune system of chickens is similar to that of mammals, they offer an attractive model system to study the effectiveness of cytokine therapy in the control of disease in livestock. This review will focus on the recent advances made in avian cytokines, with a particular focus on their assessment as therapeutic agents and vaccine adjuvants. PMID- 11943314 TI - A single amino acid deletion in the antigen binding site of BoLA-DRB3 is predicted to affect peptide binding. AB - Two bovine MHC class II alleles, BoLA-DRB3*0201 and BoLA-DRB3*3301, contain a three base pair deletion which results in the deletion of a lysine (K beta 65) in the antigen recognition site (ARS). Modelling of BoLA-DRB3*0201 with the conserved lysine K beta 65 and BoLA-DRB3*0201 without K beta 65 indicated that this deletion altered the peptide specificity of the ARS, and may impact on the immune response. To test this hypothesis, the presence of K beta 65 was analysed in a sample of cattle vaccinated with the commercial cattle tick vaccine (TickGARD). Homozygous deletion of K beta 65 was significantly associated with high response to TickGARD (P<0.05). Screening of the TickGARD antigen identified a potential T cell epitope that is recognised better by animals that are homozygous for the K beta 65 deletion. This study provides evidence that changes in the ARS of MHC class II molecules may be associated with the well recognised animal to animal variation in magnitude of vaccine response. PMID- 11943315 TI - Increase in milk metalloproteinase activity and vascular permeability in bovine endotoxin-induced and naturally occurring Escherichia coli mastitis. AB - An endotoxin-induced mastitis model was used to study permeability changes associated with increased milk matrix metalloproteinase (MMP) activity in early inflammation. One quarter of two cows was inoculated with endotoxin (Escherichia coli 055:B5). Blood, milk, and whey were collected before and repeatedly after inoculation for 48 h. The profile and amounts of gelatinolytic MMPs were determined by zymography; gelatinase A (72 kD MMP-2) and gelatinase B (92 kD MMP 9) were identified by Western immunoblotting. Bovine serum albumin (BSA) and trypsin inhibitor capacity (TIC) were used as markers of capillary permeability with parallel examination of neutrophil penetration from blood to milk. Five clinical E. coli mastitis milk samples and five milk samples from cows with healthy udders were analyzed to detect whether increased levels of gelatinolytic MMP-2 and MMP-9 have a role in naturally occurring mastitis with endotoxin involvement. Milk MMP levels increased 2h after the endotoxin challenge. Both MMP 2 and MMP-9 were involved in this early proteolytic event. These increased MMP levels are associated with increased capillary permeability, evidenced first by the penetration of small molecular weight proteins, such as BSA and TIC. Later, 6 12h post endotoxin inoculation, neutrophilic leucocytes also entered the site, as they require larger tissue damage in basal membrane and interstitial tissue structures than BSA and TIC to extravasate. In naturally occurring disease, increased MMP-2 and MMP-9 levels were detected in milk. Thus, gelatinases, especially MMP-2, are involved in the early degradation of the blood-milk barrier, which precedes the penetration of blood-derived cellular components into milk in endotoxin-induced mastitis. In the future, measuring MMP in milk/whey might be a useful tool for diagnosing early mastitis. PMID- 11943316 TI - Cytokine induction in pulmonary airways of horses with heaves and effect of therapy with inhaled fluticasone propionate. AB - Work in humans and laboratory animals has identified a central role for cytokines and chemokines in development and persistence of lower airway inflammation. The objectives of this study were to determine interleukin (IL)-1 beta, IL-2, IL-4, IL-5, IL-6, IL-8, IL-10, interferon (IFN)-gamma and tumor necrosis factor (TNF) alpha induction in bronchoalveolar lavage (BAL) of control horses and horses with heaves both during remission and exacerbation of the disease, and to determine the effect of therapy with inhaled fluticasone propionate on the cytokine profile of horses with heaves. IL-1 beta and TNF-alpha mRNA expression was significantly higher in horses with heaves after exposure to moldy hay compared to either values obtained during clinical remission or to healthy controls. IL-8 mRNA expression and protein concentrations were significantly higher in horses with heaves than in controls. Both IL-4 and IFN-gamma mRNA expression was increased at various times in heaves-susceptible horses compared to controls. IL-2, IL-5 and IL-10 mRNA expression was not detected in BAL cells of either group. Therapy with inhaled fluticasone propionate after induction of a severe heaves exacerbation resulted in complete resolution of clinical signs, normalization of pulmonary function tests, and significant decrease in BAL neutrophilia. This was associated with a significant decrease in IL-4 mRNA expression and increase in IFN-gamma/IL 4 ratio in horses with heaves. These results demonstrate the clinical efficacy of inhaled fluticasone propionate for the treatment of heaves and suggest a role for cytokines in the development of lower airway inflammation in heaves-susceptible horses. PMID- 11943317 TI - Involvement of cytolytic and non-cytolytic T cells in the control of feline immunodeficiency virus infection. AB - The appearance of non-cytolytic T cells that suppressed feline immunodeficiency virus (FIV) replication in vitro, and FIV-specific cytotoxic T cell (CTL) responses was compared in a group of seven, specific pathogen free (SPF) domestic cats following primary infection with the Glasgow(8) isolate of FIV (FIV(GL-8)). FIV proviral burdens were quantified in the blood and lymphoid tissues by real time PCR. Non-cytolytic T cell suppression of FIV replication was measured by co cultivating lymphoblasts prepared from the cats at different time-points during infection with FIV-infected MYA-1 cells in vitro. Non-cytolytic suppressor activity was detected as early as 1 week after infection, and was evident in all the lymphoid tissues examined. Further, this activity was present in subpopulations of T cells in the blood with normal (CD8(hi)) or reduced (CD8(lo)) expression of the CD8 molecule, and temporal modulations in non-cytolytic suppressor activity were unrelated to the circulating CD8(+) T cell numbers. Virus-specific CTL responses, measured by (51)Cr release assays, were not detected until 4 weeks after infection, with the emergence of FIV-specific effector CTLs in the blood. Throughout infection the response was predominantly directed towards FIV Gag-expressing target cells, and by 47 weeks after infection CTL responses had become localised in the lymph nodes and spleen. The results suggest that both non-cytolytic T cell suppression of FIV replication and FIV specific CTL responses are important cellular immune mechanisms in the control of FIV replication in infected asymptomatic cats. PMID- 11943318 TI - F4 receptor-independent priming of the systemic immune system of pigs by low oral doses of F4 fimbriae. AB - Oral administration of F4 fimbriae of Escherichia coli induces intestinal mucosal immune responses in F4 receptor-positive (F4R(+)) pigs, but not in F4R(-) pigs. We examined whether F4 fimbriae in F4R(-) animals behave like a food antigen and can induce oral tolerance. Therefore, F4R(+) and F4R(-) pigs were fed 2mg of F4 and challenged i.m. to evaluate the effect of oral F4 on the systemic immune system. As control antigen, two different oral doses (2 and 600 mg) of OVA were used. Thirty days after the i.m. OVA challenge, the OVA-specific serum IgG titre in 600 mg-fed pigs was lower than that in non-fed animals, indicating that tolerance was induced. Conversely, in the 2mg-fed pigs a rapid increase of OVA specific IgG with higher titres than those in non-fed pigs was seen following challenge, indicating a priming of the systemic immune system. A similar priming was seen in both F4-fed F4R(-) and F4R(+) pigs. Upon challenge, non-fed pigs displayed a primary immune response with a slow increase of F4-specific serum IgG, whereas F4-fed F4R(-) and F4R(+) pigs showed secondary responses with a rapid increase of serum IgG. This was expected in F4R(+) pigs, as in these animals oral F4 induces F4-specific antibody-secreting cells in the spleen, suggesting a priming of the systemic immune system. However, also the F4-fed F4R( ) pigs displayed a secondary response, despite the failure to detect a response upon oral F4 administration. These findings suggest that the F4 antigen, at a dose of 2 mg, behaves like a common food antigen in F4R(-) pigs, namely it induces a systemic priming. PMID- 11943319 TI - Pathological and immunological changes after challenge infection with Pasteurella multocida in naive and immunized calves. AB - Challenge infections of calves with Pasteurella multocida were established to characterize the local inflammatory response and determine the effect of previous exposure to live bacteria on the post-challenge immune response. Experimental infections were established by intratracheal inoculation of P. multocida in both naive calves and calves that had been previously vaccinated with two subcutaneous (s.c.) injections of live bacteria. Histological, immunohistological and cytokine expression studies were performed on bronchoalveolar lavage (BAL) samples, lung parenchymal tissues and lung lymph nodes (LN). In comparison to uninfected control animals in which no lung lesions were observed, a patchy to confluent bronchopneumonia was observed following infection of naive calves, characterized by abscess formation, haemorrhage, oedema and suppurative consolidation. Cellular analysis following infection of naive animals was characterized by an influx of neutrophils in the BAL, with macrophages and dendritic cells observed in the lesion perimeter. A significant increase in the number of CD8(+) blasts expressing MHC (major histocompatibility) II was also observed in the BAL of infected calves. Decreased expression of interleukin (IL)-1 beta and increased expression of IL-8 compared to naive unchallenged controls was apparent in lung LN. In comparison, a more limited pathology was observed in vaccinated animals post-challenge, indicating partial protection conferred by the s.c. immunization with live bacteria. Studies of vaccinated animals showed the presence of bronchial-associated lymphoid tissue (BALT) in the lung tissue and an increase in the number of B-cells and CD4(+) T-cells expressing MHCII in the lung LN after challenge. In contrast to primary infection, there was no significant influx of neutrophils in the BAL. Instead, a population of newly recruited monocytes/macrophages was observed. Increased IL-2 expression and decreased IL-8 expression was observed in the LN, while IL-1 beta expression was not detected. The reduced neutrophil and increase monocyte response in the vaccinated calves may be associated with significant changes in the gamma delta T lymphocyte population in the BAL. PMID- 11943320 TI - Is AZT/3TC therapy effective against FIV infection or immunopathogenesis? AB - In vitro and in vivo prophylactic and therapeutic efficacy of AZT/3TC treatment was evaluated against feline immunodeficiency virus (FIV) infection. In vitro studies utilized FIV-infected peripheral blood mononuclear cells (PBMCs) or FIV infected T-cell lines treated with AZT (azidothymidine) alone, 3TC alone, or AZT/3TC combination and tested for anti-FIV activity and drug toxicity. AZT/3TC combination had additive to synergistic anti-FIV activities in primary PBMC but not in chronically infected cell lines. In vivo studies consisted of four treatment groups (n=15) of SPF cats receiving AZT/3TC combination (5-75 mg/kg/drug PO BID for 8 or 11 weeks) and one control group (n=9) receiving oral placebo. Group I (n=6, 150 mg/kg/drug/day) was treated starting 3 days pre-FIV inoculation, whereas Group II (n=3, 150 mg/kg/drug/day) and Group III (n=3, 100 mg/kg/drug/day) treatments were simultaneous with FIV inoculation. Group IV treatment (n=3, 100 mg/kg/drug/day) was initiated 2 weeks post-FIV inoculation. All cats were monitored for drug toxicity and FIV infection. Eighty-three percent of cats in Group I and 33% of cats in Groups II and III were completely protected from FIV infection. A significant delay in infection and antibody seroconversion was observed in all unprotected cats from Groups I, II and III. Group IV cats had only a slight delay in FIV antibody seroconversion. Adverse drug reactions (anemia and neutropenia) were observed at high doses (100-150 mg/kg/drug/day) were reversible upon lowering the dose (20 mg/kg/drug/day). In contrast, AZT/3TC treatment had no anti-FIV activity in chronically infected cats. Furthermore, severe clinical symptoms caused by adverse drug reactions were observed in some of these cats. Overall, AZT/3TC treatment is effective for prophylaxis but not for therapeutic use in chronically FIV-infected cats. PMID- 11943321 TI - IgE enhances Fc epsilon RI expression and IgE-dependent TNF-alpha release from canine skin mast cells. AB - The role of IgE on mast cell (MC) activation is well known. Recent studies have demonstrated that IgE also has the ability to up-regulate the high affinity IgE receptor (Fc epsilon RI) on the surface of human and murine MC, leading to an increased production of cytokines and chemokines. In the present study, we have examined the influence of IgE levels on Fc epsilon RI expression, and its consequences on TNF-alpha production from canine skin MC. Mature MC were enzymatically dispersed from the skin biopsies of 6-8 dogs and were cultured for up to 5 days in medium supplemented with recombinant canine stem cell factor (SCF) (6 ng/ml), in the presence of increasing serum IgE concentrations (ranging from 0 to 80 microg/ml). Subsequently, skin MC were activated with anti-IgE, and TNF-alpha concentration was assessed 5h post-activation by a cytotoxic bioassay. Fc epsilon RI receptors were identified in MC surface by flow cytometry. MC cultured for up to 5 days in the presence of high serum IgE concentration (8 microg/ml) produced twice the quantity of TNF-alpha than MC cultured in the absence of serum IgE, in response to stimulation with anti-IgE. Moreover, the percentage of Fc epsilon RI-positive skin cells was found to be approximately double in cells cultured with serum IgE compared to that cultured in the absence of IgE, following saturation of IgE receptors. These results suggest that, as found in human and murine MC, IgE may induce an up-regulation of the Fc epsilon RI density and an enhancement in the secretory activity of canine skin MC. This study could be of great interest in designing new therapeutic strategies for controlling MC activation in inflammatory and allergic processes. PMID- 11943322 TI - Development of an ELISA for bovine IL-10. AB - The objective of the study was to develop an assay for bovine IL-10 that could be applied to analyses of immune responses and advance understanding of a variety of diseases of cattle. Recombinant bovine IL-10 (rbo IL-10) was transiently expressed in Cos-7 cells and shown to inhibit the synthesis of IFN gamma by bovine cells stimulated with antigen in vitro. Mice were immunised with a plasmid containing a cDNA insert encoding rbo IL-10 and inoculated with rbo IL-10. A number of monoclonal antibodies (mAb) were generated that reacted with rbo IL-10 in an ELISA. Some of these mAb neutralised the ability of rbo IL-10 to inhibit IFN gamma synthesis by antigen-stimulated bovine cells. A pair of mAb was identified that together could be used to detect both recombinant and natural bovine IL-10 present in supernatant of PBMC stimulated with ConA. A luminescent detection method was applied to the ELISA making it more sensitive. Using this method native IL-10 was detected in supernatants of PBMC, diluted blood and undiluted blood from cattle immunised with Mycobacterium bovis BCG or ovalbumin and incubated in vitro with antigen indicating the applicability of the assay to a number of in vitro culture systems. PMID- 11943323 TI - IgE is present on peripheral blood monocytes and B cells in normal dogs and dogs with atopic dermatitis but there is no correlation with serum IgE concentrations. AB - Blood was collected from 29 dogs, 14 with atopic dermatitis (AD) and 15 controls. Total serum IgE was quantitated. Peripheral blood monocytes were harvested and labeled with leucocyte markers and anti-canine IgE before analysis by flow cytometry. There was no statistically significant difference between the atopic and control groups when the mean number of cells in the monocyte (CD14), antigen presenting cell (CD1c) or B cell (CD21) populations were examined. However, the variation in cell numbers was significant and much greater in the atopic group for CD1c and CD14 labeled cells. The mean percentage of double labeled cells, CD1c/IgE and CD14/IgE was significantly lower in the atopic population compared with the controls. More variation was observed in the numbers of monocytes of atopic dogs (CD14/IgE) and antigen presenting cells (CD1c/IgE) of control dogs. The mean percentage of B cells expressing IgE was 65 and 51% in the atopic and control groups respectively which is greater than that reported in humans. There was no statistically significant difference. Total serum IgE concentrations were similar in each group and did not correlate with cell bound IgE in any of the leucocyte populations studied. Canine AD is associated with more variability in circulating monocyte numbers and lower numbers of monocytes expressing IgE than control dogs. Unlike in humans, there is no correlation between circulating and cell bound IgE. Furthermore, high levels of IgE in the dog may be related to a greater number of B cells in the circulation committed to IgE production. PMID- 11943324 TI - Partial cDNA sequences of bovine CD72 and CD166/ALCAM, ligands for SRCR-family accessory molecules CD5 and CD6. AB - Accessory/co-stimulatory molecules on the surface of T cells are capable of regulating activation signals. Two of these, CD5 and CD6, are molecules from the scavenger receptor cysteine rich (SRCR) superfamily. Partial sequences for the ligands of these molecules, known as CD72 and CD166 (or ALCAM), respectively, are provided for Bos taurus in this communication. Using highly conserved regions between the corresponding human and mouse genes, primers were designed and reverse transcription polymerase chain reaction was used to generate cDNA from bovine PBMC RNA. cDNA clones of several hundred base pairs in length were created and sequenced. The results showed 81% homology between bovine and human CD72 nucleotide sequences and 93% homology for the CD166 sequences. Similar levels of homology are seen between the corresponding human and mouse cDNA sequences. PMID- 11943325 TI - Regulation of adhesion molecules on circulating neutrophils during coliform mastitis and their possible immunomodulation with drugs. AB - Fast neutrophil diapedesis has been demonstrated to be critical in coliform mastitis and is determining for the severity of infection. Leukocyte adhesion molecules play a pivotal role in neutrophil recruitment. Two families of cell surface proteins help to regulate the adherence of neutrophils to vascular endothelium: selectins and beta(2)-integrins. Both classes of leukocyte adhesion molecules are reviewed in the context of their dynamic expression around parturition and during acute coliform mastitis. Their potential modulation by commonly used drugs and the therapeutic implications during acute coliform mastitis are also discussed. PMID- 11943327 TI - Production and in vivo testing of a recombinant bovine IL-12 as an adjuvant for Salmonella typhimurium vaccination in calves. AB - A recombinant bovine interleukin-12 (boIL-12) that contains a histidine hexamer, rboIL-12His, was produced, purified and administered to calves. We first tried the purification of heterodimer IL-12 from a mixture of p40 homodimer, p40 monomer, and p40-p35 heterodimer with a p35 subunit tagged with a histidine hexamar at its C-terminal (p35His). A recombinant baculovirus expressing p35His was generated and used for superinfection with a recombinant baculovirus expressing p40 subunit. The expressed subunits, p40 and p35His, were assembled into a 70kDa heterodimer in insect cells, released into culture medium, and then purified using a nickel chelate column. The purified rboIL-12His was bioactive for induction of IFN-gamma in bovine peripheral blood mononuclear cells (PBMCs) in vitro. The purified rboIL-12His was then administered to calves with inactivated Salmonella Typhimurium (ST). When sera were assayed by ELISA, specific anti-ST IgG1 antibodies were detected in all ST immunized calves, but, specific anti-ST IgG2 antibodies were detected only in calves administered ST along with rboIL-12His, indicating a possible switch to a Th1 response. Administration of commercially available Salmonella vaccine did not elicit IgG2 antibodies in calves. These results suggest that co-administration of IL-12 with inactivated ST cells could induce a Th1-type response in calves. PMID- 11943326 TI - Porcine interleukin-12 fusion protein and interleukin-18 in combination induce interferon-gamma production in porcine natural killer and T cells. AB - A pcDNA3 vector containing a gene encoding a porcine interleukin-12 (poIL-12) fusion protein was constructed, with the p40 chain and its signal peptide positioned first, followed by a linker and the p35 domain. When expressed in COS cells, secreted poIL-12 fusion protein showed high activity in terms of ability to induce interferon-gamma (IFN-gamma) production in porcine peripheral blood mononuclear cells (PBMCs) in vitro. The IFN-gamma production induced by poIL-12 fusion protein, as well as heterodimeric poIL-12 and human IL-12, was markedly dependent on the presence of human IL-18 (huIL-18). Furthermore, huIL-18 showed a dose-dependent induction of IFN-gamma production in PBMC in the presence of a constant concentration of huIL-12. A marked synergism between poIL-12 and IL-18 was consequently observed in poPBMC. The actual IFN-gamma producing cells were identified as probable NK cells (about 30%) and T lymphocytes (about 70%), using flow cytometry. Furthermore, a histidine-tagged poIL-12 fusion protein was expressed in Drosophila melanogaster Schneider 2 cells, using a modified pMT/V5 His vector lacking the V5 epitope. Such poIL-12 fusion protein was easily purified using Ni-NTA agarose and retained high biological activity. PMID- 11943328 TI - CpG-oligodinucleotides as an effective adjuvant in pigs for intramuscular immunizations. AB - In this study, the effect of two oligodeoxynucleotide (ODN) sequences 5'GCT-AGA CGT-TAG-CGT-3' (CpG-ODN) and 5'-GCT-AGA-GCT-TAG-GCT-3' (GpC-ODN) on the antigen specific antibody and cellular immune response after intramuscular immunizations with OVA was analyzed in pigs. Pigs immunized with OVA supplemented with these ODNs showed a significantly enhanced primary antibody response in comparison with the control group which received OVA without ODN. This enhanced primary antibody response appeared ODN-sequence-independent as similar effects were seen in both ODN-groups. The OVA-specific antibody titers obtained after a single injection of antigen combined with either of both ODNs were as high as the titers in the control group after two injections. Furthermore, the ODN-supplemented animals showed significantly higher OVA-specific IgA antibodies in their saliva and nasal secretions at some time points after the first immunization. Proliferation assays showed that CpG- as well as GpC-ODN significantly enhanced the antigen-specific as well as the mitogen-induced proliferation in different lymphoid tissues. Furthermore, 48h after the third immunization the CpG-group showed a significantly decreased IL-6 mRNA expression in cells of the local draining lymph node but no significant difference in TGF-beta (Th3-like) and IL-10 (Th2-like). The ODN injected animals showed the tendency to have higher IFN-gamma (Th1-like) mRNA-expression in comparison with the control group. To our knowledge, these are the first in vivo studies in pigs, which demonstrate the appropriateness of CpG ODN as immunostimulating adjuvants in vaccines for farm animals. PMID- 11943329 TI - Feline interleukin-8 expression in peripheral blood mononuclear cells induced by egg white derivatives. AB - Feline peripheral blood mononuclear cells (PBMC)-derived chemotactic factor induced by egg white derivatives (EWD) treatment was analyzed at the protein and messenger ribonucleic acid (mRNA) level. EWD itself was not active chemotactic for feline peripheral blood polymorphonuclear cells (PMN). But chemotaxis of PMN was enhanced by either culture supernatant from PBMC treated with EWD or human recombinant (hr) interleukin (IL)-8. Both hr IL-8 and the culture supernatant from PBMC treated with EWD yielded a distinct band, molecular weight of 6-8kDa, in sodium-dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) with 15% loading gel. Therefore, to identify this chemotactic factor, culture supernatant from PBMC treated with EWD was partially purified by anion exchange chromatography on diethylaminoethyl (DEAE)-Sepharose CL-6B and concentrated by ultrafiltration. Only the fraction, which was eluted with 0.3M NaCl, showed a high concentration of total protein and also enhanced the chemotactic activity of PMN. This activity was thereafter designated as eluate. The chemotactic activity of eluate was inhibited by anti-hr IL-8 polyclonal antibody (pAb). A single protein band with 6-8kDa was shown in both the eluate and hr IL-8 when analyzed by SDS-PAGE and Western blotting using anti-hr IL-8 pAb, suggesting that the chemotactic factor for feline PMN is IL-8, 6-8kDa, produced by PBMC treated with EWD. The physicochemical characteristics of eluate were stable in heated (60-100 degrees C), acid (pH 3.0), and alkaline (pH 9.0) conditions. The eluate under these conditions also showed a distinct band in molecular weight of 6-8kDa in SDS PAGE and Western blotting and was very active in chemotactic activity of PMN.IL-8 mRNA gene expression on feline PBMC was analyzed by reverse transcriptase polymerase chain reaction (RT-PCR) assay using a series of oligonucleotides, each 22 mer, derived from feline IL-8. Feline IL-8 mRNA showed low level in 3-h incubation without EWD, but it was increased in a dose-dependent manner by addition of EWD. Following EWD (10 microg/ml) treatment, IL-8 mRNA expression was rapidly increased up to 6h and decreased by 12h although it was not expressed in freshly prepared PBMC. This study strongly suggested that immunoenhancing effect of EWD on chemotactic response of PMN is mediated by feline IL-8, 6-8kDa, produced by PBMC stimulated with EWD. In addition, the expression of feline IL-8 mRNA on PBMC is increased when stimulated with EWD. PMID- 11943330 TI - Development of a rapid in vitro protein refolding assay which discriminates between peptide-bound and peptide-free forms of recombinant porcine major histocompatibility class I complex (SLA-I). AB - The extracellular domains of swine leukocyte antigen class I (SLA-I, major histocompatibility complex protein class I) were cloned and sequenced for two haplotypes (H4 and H7) which do not share any alleles based on serological typing, and which are the most important in Danish farmed pigs. The extracellular domain of SLA-I was connected to porcine beta2 microglobulin by glycine-rich linkers. The engineered single-chain proteins, consisting of fused SLA-I and beta2 microglobulin, were overexpressed as inclusion bodies in Escherichia coli. Also, variants were made of the single-chain proteins, by linking them through glycine-rich linkers to peptides representing T-cell epitopes from classical swine fever virus (CSFV) and foot-and-mouth disease virus (FMDV). An in vitro refold assay was developed, using a monoclonal anti-SLA antibody (PT85A) to gauge refolding. The single best-defined, SLA-I restricted porcine CD8(+) T-cell epitope currently known is a 9-residue peptide from the polyprotein of CSFV (J. Gen. Virol. 76 (1995) 3039). Based on results with the CSFV epitope and two porcine haplotypes (H4 and H7), the in vitro refold assay appeared able to discriminate between peptide-free and peptide-occupied forms of SLA-I. It remains to be seen whether the rapid and technically very simple in vitro refold assay described here will prove generally applicable for the screening of virus-derived peptides for SLA-I binding. PMID- 11943331 TI - Effect of different adjuvants on the immune responses of cattle vaccinated with Mycobacterium tuberculosis culture filtrate proteins. AB - The development of improved vaccines for bovine tuberculosis is urgently required as a cost effective solution for control and eventual eradication of tuberculosis in domestic animals. Studies in small animal models of tuberculosis have shown that vaccination with culture filtrate proteins (CFP), prepared from Mycobacterium tuberculosis or M. bovis, can induce cellular immune responses and confer a level of protection against aerogenic challenge with virulent mycobacteria. As a first step in the development of a mycobacterial CFP vaccine for protection of cattle against bovine tuberculosis, the immune responses of cattle vaccinated with short-term culture filtrate proteins (ST-CFP) from M. tuberculosis and formulated with different adjuvants were compared with those vaccinated with bacille Calmette-Guerin (BCG). The adjuvants included dimethyldioctyldecyl ammonium bromide (DDA), diethylaminoethyl (DEAE)-dextran, and ST-CFP adsorbed onto polystyrene beads. Vaccination with ST-CFP/DEAE-dextran induced high levels of interleukin-2 (IL-2) but low levels of interferon-gamma (IFN-gamma) from whole-blood cultures stimulated with M. tuberculosis ST-CFP in comparison with the strong IFN-gamma and IL-2 responses induced after vaccination with BCG. ST-CFP/DEAE-dextran also induced a strong antigen-specific immunoglobulin antibody response with both immunoglobulin G1 (IgG1) and IgG2 isotypes. Vaccination with ST-CFP/beads induced a weak IgG1-biased antibody response but no IFN-gamma or IL-2 response. DDA did not induce significant immune responses in animals vaccinated with ST-CFP. In comparison to the moderate delayed-type hypersensitivity (DTH) responses induced by vaccination with subcutaneous BCG, none of the ST-CFP vaccines induced a significant DTH response to either M. tuberculosis ST-CFP or bovine purified protein derivative (PPD). While the ST-CFP vaccines used in this study have not induced strong antigen specific cellular immune responses in cattle comparable to those induced by BCG, they are immunogenic in cattle and it may be possible to overcome this problem by using adjuvants that more effectively promote IFN-gamma responses in this species. PMID- 11943332 TI - Antibody response to a fragment of the protein G of VHS rhabdovirus in immunised trout. AB - A fragment (called frg#11, amino acids, aa 56-110) of the protein G (pG) of viral haemorrhagic septicaemia virus (VHSV) was designed after previous results showed it to be recognised by approximately 40% of the trout immunised to VHSV [Dis. Aquat. Organ. 34 (1999) 167]. frg#11 was then cloned, expressed, purified and used to study the production of antibodies to its epitopes in trout immunised to VHSV. Anti-frg#11 trout antibodies could be detected in serum from individual trout surviving VHSV exposure, immunised by injection with purified VHSV or DNA immunised with its pG gene whereas it was not detected in non-infected and non immunised trout. The trout serum antibodies which reacted more strongly by ELISA using solid-phase frg#11 (continuous or linear epitopes on the sequence of the pG) had the lowest VHSV-neutralising activity (epitopes which are pG conformation dependent). Because antibodies recognising continuous as well as conformation dependent epitopes of the pG seem to be involved in protective trout immunological responses to VHSV, the estimation of anti-frg#11 antibodies could help to the dissection of the complex trout antibody response to VHSV infections. In addition, these preliminary results suggest that the determination of anti frg#11 antibodies might also be used to complement in vitro viral neutralising assays which seem to be restricted to pG conformation-dependent epitopes. PMID- 11943333 TI - Immunostimulation in the rainbow trout (Oncorhynchus mykiss) following intraperitoneal administration of Ergosan. AB - The present work provides information concerning the immunostimulatory activity of Ergosan, an algal based product, injected intraperitoneally in the rainbow trout (Oncorhynchus mykiss). Ergosan is composed of 0.002% unspecified plant extract, 1% alginic acid from Laminaria digitata, and 98.998% algal based carrier. Migration of leucocytes into the peritoneal cavity was stimulated at doses > or =1 mg ml(-1). A single dose of 1mg significantly augmented the proportion of neutrophils, degree of phagocytosis, respiratory burst activity and expression of interleukin-1beta (IL-1beta), interleukin-8 (IL-8) and one of the two known isoforms of trout tumour necrosis factor-alpha (TNF2) in peritoneal leucocytes at 1 day post-injection. Humoral immune parameters were less responsive to intraperitoneal Ergosan administration, with complement stimulation only evident in the 1mg treated group at 2 days post-injection. Antiprotease and lysozyme activity were unaffected by Ergosan over a 7-day time period at the doses examined. PMID- 11943334 TI - Recombinant bovine soluble CD14 sensitizes the mammary gland to lipopolysaccharide. AB - Standard therapies including administration of potent antibiotics, aggressive fluid resuscitation and metabolic support have not been successful in relieving symptoms and reducing mortality associated with acute coliform mastitis. It is important to understand the pathophysiological response of the mammary gland to coliform infections when designing preventive or therapeutic regimens for controlling coliform mastitis. Our laboratory has previously shown that macrophages and polymorphonuclear neutrophils in milk express CD14 on their cell surface. In this study, we found that soluble CD14 (sCD14) is present in milk whey as a 46kDa protein reacted with anti-ovine CD14 antibody. Additional functional studies found that: (1) under serum-free condition, complexes of LPS recombinant bovine soluble CD14 (rbosCD14) induced activation of mammary ductal epithelial cells (as measured by changes in interleukin-8 (IL-8) mRNA level by competitive RT-PCR) at low concentrations of LPS after 6 or 24h incubation (1 1000ng/ml), whereas LPS alone did not induce activation of mammary ductal epithelial cells at the same concentrations, and (2) intramammary injection of low concentrations of LPS did not increase concentration of leukocytes in milk. In contrast, LPS-rbosCD14 complex containing the same concentration of LPS increased the concentration of leukocytes in the injected mammary gland at 12 and 24h post-injection. These results indicate that rbosCD14 sensitizes mammary epithelial cells to low concentrations of LPS in vitro and in vivo. Endogenous sCD14 in milk may be important in initiating host responses to Gram-negative bacterial infections. PMID- 11943335 TI - Alterations in the INK4a/ARF locus and their effects on the growth of human osteosarcoma cell lines. AB - Two different proteins, p16(INK4a) and p14(ARF), encoded by the INK4a/ARF locus play important roles in the RB and p53 pathways, respectively. This study was performed to determine genetic and epigenetic alterations in the INK4a/ARF locus and their effects on the growth of osteosarcoma. Among six cell lines examined, both p16(INK4a) and p14(ARF) exons were homozygously deleted in two cell lines, MG63 and HOS, and both p16(INK4a) and p14(ARF) promoters were methylated in one cell line, U2OS. Wild-type mRNA and proteins for p16(INK4a) and p14(ARF) were expressed in three other cell lines, SaOS2, HuO9, and G292. Transfection studies were performed using two cell lines, U2OS and MG63. Both the RB and p53 genes were wild types in U2OS, whereas p53 but not RB was mutated in MG63. Both p16(INK4a) and p14(ARF) suppressed the growth of U2OS, whereas p16(INK4a) but not p14(ARF) suppressed the growth of MG63. p53 only did not suppress the growth of MG63 either; however, coexpression of p14(ARF) with p53 increased the fraction of the G0/G1 phase in MG63 cells. The data presented here demonstrate the importance of genetic and epigenetic alterations in the INK4a/ARF locus for the growth of osteosarcoma and thus will be useful to further understand the biologic behavior of osteosarcoma in association with the defects in the p53 and RB pathways. PMID- 11943336 TI - Organophosphate-based pesticides and genetic damage implicated in bladder cancer. AB - Organophosphate-based pesticides have been associated with pathology and chromosomal damage in humans. There are also epidemiologic links with cancer. The few screening tests for low-level occupational exposure are of doubtful sensitivity; this investigation evaluated four methods. Blood samples were studied from 10 farmers before and after occupational exposure to organophosphate based pesticides and five unexposed controls. The standard cholinesterase test was insensitive to the exposure (P=0.815). However, a significant increase in Howell-Jolly bodies within erythrocytes was observed (P=0.001). Cytogenetic studies on routine and aphidicolin-induced blood cultures revealed that following organophosphate exposure the total number of gaps and breaks on human chromosomes was significantly increased (P=0.004 and P=0.0006, respectively). We concluded that Howell-Jolly body and fragile site analysis were sensitive indicators of nuclear damage resulting from low-level occupational exposure to organophosphate. Such nuclear damage could be implicated in carcinogenesis. The development of bladder cancer is one such example. PMID- 11943337 TI - Clinical implications of PRAME gene expression in childhood acute myeloid leukemia. AB - The expression of the PRAME gene (preferentially expressed antigen of melanoma) was measured by quantitative reverse transcriptase polymerase chain reaction in 50 children with newly diagnosed acute myeloid leukemia (AML), three samples of CD34(+) stem cells, six bone marrow samples, and 10 peripheral blood samples of healthy donors, as well as three AML cell-lines (KG-1, U937, and HL-60). Eight patients were also analyzed in relapse. Contrary to previous reports, we could show that the PRAME gene is expressed by CD34(+) stem cells. This might constitute a problem in using PRAME for tumor immunotherapy. Overexpression of PRAME was found in 62% (n=31) of our patients. The rates of overall and disease free survival in this group were higher than in patients with no or low expression (P<0.05). PRAME expression was negatively correlated to the white blood cell count at diagnosis (P<0.05) and significantly higher in patients with t(8;21). The levels of expression at diagnosis corresponded with those at relapse (P<0.001) and increased levels could be found prior to the relapse in one patient who was regularly monitored. Our results suggest that the expression of PRAME is an indicator of favorable prognosis and could be a useful tool for monitoring minimal residual disease in childhood AML. PMID- 11943338 TI - Ossifying fibromyxoid tumor of soft parts. Cytogenetic findings. AB - Ossifying fibromyxoid tumor (OFMT) of soft parts is a recently described, rare but morphologically distinctive soft tissue tumor. The histogenesis of this lesion remains uncertain, although several immunohistochemical and ultrastructural features suggest that it is an unusual neural tumor, possibly of Schwann cell origin. We report here a case of a malignant variant of OFMT that occurred in the foot of a 52-year-old man. The karyotype of a pulmonary metastasis exhibited the following complex numeric and structural aberrations:72 approximately 74,XXY, 5,+6,+del(8)(p21),del(9)(p22),+10,der(11)t(3;11)(p21;p15),del(12) (q13),der(13)t(5;13)(q13;q34),+18,+19,+20,-22 [cp10]. A kidney metastasis exhibited the following karyotypic abnormalities: 46,XY,add(3)(p11),+der(3)t(3;?;11)(3qter-->3p11::?::11q13-->11qter), 5,del(8)(p21),add(9)(q22),del(9)(p22),der(11)t(3;11)(p21;p15),del(12)(q13),+der(1 )t(5;13) (q13;q34),-22. To our knowledge, this is the first reported case of OFMT in which clonal chromosomal aberrations have been shown. PMID- 11943340 TI - Cytogenetic characterization of complex karyotypes in seven established melanoma cell lines by multiplex fluorescence in situ hybridization and DAPI banding. AB - We report the use of multiplex fluorescence in situ hybridization (M-FISH) to resolve chromosomal aberrations in seven established melanoma cell lines with hypotriploid to hypertetraploid complex karyotypes. By simultaneous identification of all human chromosomes in single FISH experiments using a set of 52 directly labeled, whole chromosome painting probes, cryptic chromosomal translocations and the origin of unclear chromosomal material in structural rearranged and marker chromosomes could be identified, refining the tumor karyotypes in all seven cell lines. The number of structural aberrations in each cell line assigned with combined M-FISH and DAPI banding analysis ranged from 15 to 45. Altogether, 275 breakpoints could be assigned to defined chromosomal regions or bands. The chromosome arms 1p, 6q, 7p, 9p, and 11q which are known to be nonrandomly associated with melanoma tumorigenesis, were frequently involved in chromosomal breaks and/or copy number changes. This study also demonstrated the practical usefulness of combining M-FISH with conventional cytogenetic banding techniques for the characterization of complex tumor karyotypes with massive genomic alterations. PMID- 11943341 TI - Primary marginal zone lymphoma of the thymus accompanied by chromosomal anomaly 46,X,dup(X)(p11p22). AB - We report a case of primary marginal zone lymphoma in the thymus of a 34-year-old woman. She was initially suspected of having a mediastinal plasmacytoma because of the presence of dominantly proliferating plasmacytic cells in a small fragment obtained by thoracoscopic biopsy, and an elevated level of serum monoclonal IgA. However, histology of the tissue obtained by a subsequent open surgical biopsy revealed diffuse proliferation of atypical monocytoid B-lymphocyte-like cells, which showed prominent plasmacytic differentiation and a close association with thymic epithelial cells consistent with the histology of a marginal zone lymphoma of the thymus. These lymphoma cells were positive for CD19, CD20, IgA, and kappa, and negative for CD5, CD10, and other T/NK-cell and myelomonocyte antigens. Both G-banded and spectral karyotyping analyses revealed the lymphoma cells carried a chromosomal anomaly, 46,X,dup(X)(p11p22). Although large cell type B-cell lymphoma in the thymus (mediastinal diffuse large B-cell lymphoma), which is categorized as a definite subtype in revised European-American classification of lymphoid neoplasms and the new World Health Organization classification, is not infrequent, primary marginal zone lymphoma of the thymus is extremely rare. To our knowledge, this is the first case report of primary marginal zone lymphoma of the thymus with a detailed chromosomal analysis. PMID- 11943339 TI - Molecular analysis of an unusual rearrangement between chromosomes 4 and 11 in adult pre-B-cell acute lymphoblastic leukemia. AB - The reciprocal t(4;11)(q21;q23) is often described in acute lymphoblastic leukemia (ALL). In some cases, variant or complex t(4;11) has been detected in ALL by cytogenetic analysis, but to our knowledge no molecular study has been reported in these variant translocations. We describe a 27-year-old woman suffering from pre-B-cell ALL with an unusual rearrangement between chromosomes 4 and 11. Because this complex rearrangement involved the 11q23 chromosome band known to be associated with poor prognosis, we performed fluorescence in situ hybridization, Southern blot, and reverse transcription polymerase chain reaction analyses. This confirmed myeloid lymphoid leukemia gene rearrangement and showed the presence of MLL/AF4 fusion transcript. These results showed the importance of molecular analysis that allowed minimal residual disease monitoring in this patient. PMID- 11943342 TI - Duplication 15q in a patient with t(8;21) acute myeloblastic leukemia (M2). AB - We present unique chromosomal abnormalities found in a patient with acute myeloblastic leukemia (AML) of French-American-British subtype M2. The patient was referred for an evaluation of chromosomal anomaly associated with AML. She was found to have an abnormal karyotype 46,XX,t(8;21)(q22;q22), and a questionable dup(15)(q15q22) in the majority of cells analyzed. Two cells had the same chromosomal anomalies plus a duplicated derivative chromosome 21 [der(21)t(8;21)(q22;q22)]. These cytogenetic findings were confirmed by fluorescence in situ hybridization utilizing the appropriate DNA probes. To our knowledge, this is the first case report of a combination of the translocation between chromosomes 8 and 21, a duplication of chromosome 15 [dup(15)(q15q22), and a duplicated derivative chromosome 21 [der(21)t(8;21)(q22;q22)]. PMID- 11943343 TI - Gastrointestinal stromal tumor in an XYY/XY male. AB - A 32-year-old patient was diagnosed with a gastrointestinal stromal tumor of the small bowel. The pathologic diagnosis was confirmed by positive immunochemistry against CD34, and against CD117, the tyrosine-kinase c-kit. We performed a karyotypic analysis on the basis of the patient's tall stature and speech difficulties. One hundred thirty-two metaphases were obtained on PHA-stimulated peripheral blood; 123 of them presented an extra chromosome Y. Fluorescence in situ hybridization using a Y satellite III probe showed the presence of a sole copy of chromosome Y in the tumor cells precluding a direct relationship between the extra chromosome Y and the initiation of the tumor. This is, to our knowledge, the second occurrence of a nonhematologic malignancy reported in this genetic disorder. A review of the malignancies observed in men with the XYY constitution is presented. PMID- 11943344 TI - Rearrangement involving chromosomes 1 and 8 in a retroperitoneal lipoma. AB - Superficial lipomas are very common benign adipose tissue tumors. In contrast, deep-seated lipomas such as retroperitoneal lipomas, are extremely rare and have to be carefully distinguished from well-differentiated liposarcomas for appropriate treatment and follow-up. We report to, our knowledge, the first cytogenetic analysis of a retroperitoneal lipoma occurring in an adult, which showed a complex rearrangement interpreted as t(1;8)(q32;q22-q23) followed by a pericentric inversion of der(8). There was no detectable rearrangement of chromosome 12, and in particular no 12q14-q15 amplification. Because rearrangements of the 8q11-q13 region involving the PLAG1 gene have been described in lipoblastoma-another kind of benign adipose tumor--we used fluorescence in situ hybridization analysis to determine in the present case the chromosomal breakpoint on 8q was located between the ETO (8q22) and COX6C (8q22 q23) genes at a great distance from PLAG1. Karyotypic analysis of additional cases of retroperitoneal lipomas will be required to assess the significance of chromosome 1 and 8 rearrangements in a continuous effort to attain a better classification of adipose tissue tumors. Of great importance is the determination of such genetic markers as additional tools for the differential diagnosis between benign and malignant forms of adipose tumors, and to avoid erroneous diagnoses. PMID- 11943345 TI - Concomitant tetrasomy 3q and trisomy 18 in CD5(-), CD13(+) chronic lymphocytic leukemia. AB - Chronic lymphocytic leukemia (CLL) associated with myeloid antigens on the surface of B neoplastic cells is a recently identified immunologic variant confined to patients with CD5 negative B-CLL. We describe the case of a 61-year old female diagnosed with CD5(-), CD13(+) B-CLL with a tetrasomy 3q revealed by fluorescence in situ hybridization analysis, in addition to trisomy 18. To our knowledge, this is the first reported case of B-CLL with this kind of cytogenetic abnormality. PMID- 11943347 TI - Common fragile sites associated with the breakpoints of chromosomal aberrations in hematologic neoplasms. AB - Fragile sites are specific regions of chromosomes prone to breakage when cells are cultured under specific conditions. These sites are divided into two classes: common and rare. Common fragile sites are expressed in all individuals at different frequencies, whereas rare ones are found only in certain individuals. Common and rare fragile sites have been shown to display a number of characteristics of instability being preferential sites for chromosomal deletions, duplications, and rearrangements. Moreover, a majority of mapped oncogenes are located at or near these fragile sites. These observations have led to the suggestion that both classes of fragile sites may play a significant role in chromosomal rearrangements involved in oncogene activation or tumor supressor gene inactivation. For these reasons, involvement of common and rare fragile sites and their relevance to specific chromosome breakpoints in cancer have received much attention. In this study, which reports on the cytogenetic findings obtained from 256 patients with chronic myelocytic leukemia, 103 with acute myelocytic leukemia, 40 with acute lymphocytic leukemia, 33 with myelodysplastic syndrome, we documented the fragile sites involved in chromosomal aberrations involving oncogenes, tumor supressor genes, and other known genes important in cell cycle regulation localized at these sites. PMID- 11943346 TI - Eosinophilic leukemia associated with t(2;5)(p23;q31). AB - Chromosomal aberrations have been reported in most malignant hematopoietic disorders such as acute or chronic myeloid leukemia, acute lymphoid leukemia, and myelodysplastic syndromes. Eosinophilic leukemia is a rare hematologic malignancy difficult to distinguish from other forms of idiopathic hypereosinophilic syndrome, so that the diagnosis is often made by exclusion, unless cytogenetic abnormalities can be demonstrated in bone marrow cells. We describe a patient with eosinophilic leukemia whose cytogenetic study shows a t(2;5)(p23;q31). Initial data could suggest a clonal eosinophilia, with an hepatosplenomegaly, severe pancytopenia, and a high level of blood and medullar eosinophilia. PMID- 11943348 TI - Trisomy 4 as the sole cytogenetic abnormality in a Waldenstrom macroglobulinemia. AB - We report a case of Waldenstrom macroglobulinemia with trisomy 4 as the sole cytogenetic abnormality. Trisomy 4 has been reported previously in Waldenstrom macroglobulinemia, but only in conjunction with multiple chromosomal aberrations. Trisomy 4 has been reported in other hematologic malignancies including acute myeloid and lymphoid leukemias. PMID- 11943349 TI - Recurrent DNA sequence copy losses on chromosomal arm 6q in capillary hemangioblastoma. AB - Capillary hemangioblastomas (CHB) of the central nervous system, the most common tumor in von Hippel-Lindau (VHL) disease, usually show mutations in the VHL tumor suppressor gene on chromosome 3p25-p26. Because little is known concerning the cytogenetic changes in these tumors, we studied 22 cases through comparative genomic hybridization to screen for DNA copy number changes in both sporadic and VHL-associated CHB. Our analysis revealed that 6 of 22 samples (27%) contained DNA copy number losses, whereas no gains were observed. The most recurrent finding was loss of chromosomal arm 6q, seen in five cases. In two of these cases also loss of chromosome 3 was noted. The third aberration observed was loss of chromosome 8, seen in one case. No differences were noted between VHL-associated and sporadic tumors, nor did the cytogenetic aberrations correlate with the clinical outcome. The loss of 6q, seen in this study and previously in other VHL associated tumors (renal cell carcinomas and pheochromocytomas) and other tumors, suggest that this chromosome area may contain tumor suppressor genes involved in the early steps of tumorigenesis. PMID- 11943350 TI - Cytogenetic findings in two new cases of hepatoblastoma. AB - We describe two cases of hepatoblastoma occurring in an 18-month-old boy and a 3 month-old girl. Cytogenetic analysis of the primary tumors revealed gain of 2q and 20 in both cases. In case 1, t(7;8;11) was seen as a secondary abnormality. Other chromosomal aberrations seen in case 2 were unbalanced t(1;1) and t(2;11), resulting in partial gains of 1q and 2q. These results support previous reports that gains of 2q and 20 and rearrangement of chromosome 1 are strongly associated with hepatoblastoma and may be essential for establishing this neoplasm. The 11q and 7q abnormalities may represent a pathway of genetic evolution associated with hepatoblastoma progression. PMID- 11943351 TI - A new case of CD7-positive acute myeloblastic leukemia with trisomy 21 as a sole acquired abnormality. PMID- 11943352 TI - Acute myelomonocytic leukemia with t(1;18) in an adult patient. PMID- 11943353 TI - Use of the green fluorescent protein to monitor the fate of Serratia entomophila causing amber disease in the New Zealand grass grub, Costelytra zealandica. AB - A series of constitutive green fluorescent protein (pGFPuv) derivatives of the bacterium Serratia entomophila (Enterobacteriaceae) were constructed, allowing the fate of cells causing amber disease ingested by the New Zealand grass grub (Costelytra zealandica, Coleoptera: Scarabaeidae) to be monitored. Examination of tissue and contents of the alimentary tract over time from ingestion, under fluorescence microscopy, revealed that the major site of S. entomophila colonisation in the grass grub is intestinal particulate matter. Visual examinations showed that wild type pathogenic strain persisted in high numbers in the grass grub intestinal tract, notably in the area of the hindgut, but the S. entomophila pADAP-free strain 5.6RC and the pADK mutant derivatives (pADK-4, -10, -13) that gave a non-feeding without gut clearance phenotype, were unable to colonise the gut. The indiscriminate colonisation of the intestinal tract particulate matter by pathogenic bacteria, rather than the colonisation of a specific site of activity, suggests that the bacterial toxins are induced and released from the bacteria while they live freely in the grass grub intestinal tract. PMID- 11943354 TI - A comparative evaluation of five typing techniques for determining the diversity of fluorescent pseudomonads. AB - Five typing methods were evaluated, utilising 63 strains of fluorescent pseudomonads, to assess their usefulness as tools to study the bacterial diversity within this complex group. The methods used were Biolog metabolic profiling, restriction fragment length polymorphism ribotyping, PCR ribotyping, and repetitive element sequence-based PCR (rep-PCR) utilising BOX and enterobacterial repetitive intergenic consensus (ERIC) primers. Cluster analysis of the results clearly demonstrated the considerable homogeneity of Pseudomonas aeruginosa isolates and, conversely, the heterogeneity within the other species, in particular P. putida and P. fluorescens, which need further taxonomic investigation. Biolog metabolic profiling enabled the best differentiation among the species. Rep-PCR proved to be highly discriminatory, more so than the other DNA fingerprinting techniques, demonstrating its suitability for the analysis of highly clonal isolates. RFLP ribotyping, PCR ribotyping, and rep-PCR produced specific clusters of P. aeruginosa isolates, which corresponded to their origins of isolation, hence we recommend these methods for intraspecific typing of bacteria. PMID- 11943355 TI - Modified enzyme activity assay to determine biofilm biomass. AB - An assay of potential exoproteolytic enzyme activity was modified to quantitatively measure the biomass of attached biofilm. The assay utilized the nonfluorescent compound L-leucine-beta-naphthylamide (LLbetaN) that becomes fluorescent when bacterial exoenzymes break the peptide bond, releasing the fluorochrome beta-naphthylamine. Fluorescence development was measured by pumping the liquid phase of a biofilm sample through a fluorescence detector and recording the detector output using a personal computer. A significant linear relationship was shown to exist between the rate of fluorescence development and the biofilm's biomass as carbon, determined using total direct cell counts, measured cell volumes and an existing relationship between cell volume and cell carbon. The technique was used to measure biofilm biomass for experiments where iron oxides were the substratum. Biofilm biomass measurements made using heterotrophic plate counts (HPCs) on R2A medium were shown to correlate well to biomass measurements made using the modified enzyme assay. The technique was shown to be sufficiently sensitive to measure biomass on samples containing little biofilm biomass, such as those exposed to free chlorine. While granular and porous media were used for the experiments presented, small biofilm coupons could easily be used to measure biofilm biomass, expanding the number of possible applications for the enzyme assay technique. PMID- 11943356 TI - Evaluation of the MicroFoss system for enumeration of total viable count, Escherichia coli and coliforms in ground beef. AB - This study was conducted to evaluate the performance of the MicroFoss system (Biosys, Ann Arbor, MI) for enumeration of total viable organisms, Escherichia coli and coliforms in ground beef. The system performance was compared to that of the USDA Bacteriological Analytical Method (BAM) reference culture methods. The correlation coefficients for the regression lines comparing the MicroFoss system detection times to the results of plate count methods for the total viable counts, coliform counts and the most probable number (MPN) method for E. coli were -0.95, -0.96 and -0.97, respectively. Tests comparing the reproducibility of data generated independently by two technicians on the same batch of samples showed no significant differences (P>0.05) in the MicroFoss detection times and culture results. The plate count methods for the total viable counts and coliform counts, and the MPN method for E. coli required 10, 11 and 22 times, respectively, the amount of time to complete tests compared to the length of time required to perform these tests using the MicroFoss system. The MicroFoss system produced reproducible data and provided a rapid and cost-efficient alternative method for enumeration of TVC, coliforms and E. coli in ground beef. PMID- 11943357 TI - Electron energy loss spectroscopy techniques for the study of microbial chromium(VI) reduction. AB - Electron energy loss spectroscopy (EELS) techniques were used to determine oxidation state, at high spatial resolution, of chromium associated with the metal-reducing bacteria, Shewanella oneidensis, in anaerobic cultures containing Cr(VI)O4(2-). These techniques were applied to fixed cells examined in thin section by conventional transmission electron microscopy (TEM) as well as unfixed, hydrated bacteria examined by environmental cell (EC)-TEM. Two distinct populations of bacteria were observed by TEM: bacteria exhibiting low image contrast and bacteria exhibiting high contrast in their cell membrane (or boundary) structure which was often encrusted with high-contrast precipitates. Measurements by EELS demonstrated that cell boundaries became saturated with low concentrations of Cr and the precipitates encrusting bacterial cells contained a reduced form of Cr in oxidation state + 3 or lower. PMID- 11943358 TI - Electro-orientation of Schizosaccharomyces pombe in high conductivity media. AB - The orientation of microbial cells may be important in cell-cell interactions within microbial consortia. As part of our research programme aimed at the construction of Artificial Structured Microbial Consortia (ASMC), we have investigated the electro-orientation of Schizosaccharomyces pombe in AC electric fields, and studied the effects of the applied frequency, voltage, and distance between the electrodes, at different medium conductivities. It is shown that the electro-orientation of S. pombe in media with conductivities similar to that of growth media is feasible using microelectrodes. Oriented growth of S. pombe can be obtained when continuously exposed to AC electric fields in growth medium over extended periods. PMID- 11943359 TI - Evaluation of redox indicators and the use of digital scanners and spectrophotometer for quantification of microbial growth in microplates. AB - The growth indicators 2,3,5-triphenyltetrazolium chloride (TTC), 2-[4-iodophenyl] 3-[4-dinitrophenyl]-5-phenyltetrazolium chloride (INT), 2,3-bis[2-methoxy-4-nitro 5-sulfophenyl]-2H-tetrazolium-5-carboxanilide inner salt (XTT), 3-[4,5 dimethylthiazol-2-yl]-2,5-diphenyltetrazolium bromide (MTT), and resazurin were tested for their ability to indicate bacterial growth/growth inhibition. Two reading devices were evaluated and compared, a microplate spectrophotometer and a digital flatbed scanner. The bacteria used in the study were cultivated in 96 wells microplates and readings were made after 24 h. The scanned pictures were analysed with a software developed in-house to generate numerical values. It was found that resazurin was difficult to use since it shifts between three colours. MTT and TTC had a high correlation between the spectrophotometer data and the data from the scanned images. The reproducibility was similar for both reading devices. In no case was there a need to resuspend the pellets before reading. Both the XTT and INT showed lower correlations. It is concluded that bacterial growth/growth inhibition can be easily and reproducibly measured from microplate cultivations with a flatbed scanner or with a microplate spectrophotometer. PMID- 11943360 TI - Detection in coal tar waste-contaminated groundwater of mRNA transcripts related to naphthalene dioxygenase by fluorescent in situ hybridization with tyramide signal amplification. AB - The ideal ecological metabolic activity assay would be applied to naturally occurring microbial populations immediately fixed in the field, and the assay would focus upon intracellular parameters indicative of a dynamic biogeochemical process. In this study, fluorescent in situ hybridization (FISH) with tyramide signal amplification (TSA) detected intracellular mRNA in bacteria. Detection sensitivity was enhanced by using a Hamamatsu color chilled CCD camera and extended exposure times. Pseudomonas putida NCIB 9816-4, a model naphthalene degrading bacterium, was used to refine the protocol. Probe Ac627BR was developed for detecting naphthalene dioxygenase (nahAc) mRNA transcripts. Only induced cells showed positive hybridization to probe Ac627BR. Results were verified by RNase A or DNase I digestion of samples prior to hybridization. When applied to field-fixed groundwater samples, the naphthalene dioxygenase mRNA probe conferred fluorescence on a subset (approximately 1%) of the cells present in the contaminated groundwater. This methodology represents progress towards achieving one of the longstanding goals of environmental microbiology: to simultaneously ascertain the identity, activity, and biogeochemical impact of individual microorganisms in situ-in soil, water, or sediment where they dwell. PMID- 11943361 TI - Non-radioactive PCR-SSCP with a single PCR step for detection of inhibitor resistant beta-lactamases in Escherichia coli. AB - A method based on PCR-SSCP has been developed to detect presumptive Inhibitor Resistant TEM (IRT) beta-lactamases in Escherichia coli. The capacity of this technique to differentiate genes from 11 control strains encoding IRT beta lactamases was evaluated with PCR products digested with RsaI. All the bla(TEM) genes studied could be distinguished by their electrophoretic mobilities. Applied to 29 epidemiologically unrelated clinical isolates of E. coli resistant to amoxicillin-clavulanate (MIC, > or =32 microg/ml), the electrophoretic mobilities of the digested bla(TEM) PCR products were identical to those of the reference bla(TEM-1A) (6 strains) and bla(TEM-1B) (18 strains) genes. The remaining five bla(TEM) PCR products displayed SSCP profiles different from those of the reference bla(TEM) genes and their nucleotide sequence identified them as bla(TEM 1C) in one strain, bla(TEM-30/IRT-2) in two strains, bla(TEM-37/IRT-8) in one strain, and bla(TEM-40/IRT-11) in one isolate. Overexpression of the wild-type bla(TEM-1) gene, as detected by high-level resistance to beta-lactams and enzyme assay, accounted for resistance in the 24 E. coli containing bla(TEM-1). We report a simple one PCR step SSCP that can be used in epidemiological studies for rapid preliminary detection of IRT beta-lactamases; identification should be confirmed by sequence data. PMID- 11943362 TI - Purification and characterization of 3,3-dihydroxyazetidine from culture medium of Bacillus mesentericus and B. subtilis. AB - A method is described for the purification of 3,3-dihydroxyazetidine (DHA), which accelerates the growth of Bifidobacterium spp., from the culture medium of Bacillus mesentericus (BM). Purification involved adsorption to an ion-exchange resin; it required less time and gave a higher yield than a previously reported method. Monitoring the inhibition of E. coli NIHJ JC-2, we searched for other strains that produced 3,3-dihydroxyazetidine. We found that not only B. mesentericus TO-A but also B. subtilis IFO13719 produced the compound of interest. PMID- 11943363 TI - A filtration, incubation and staining reactor including a new protocol for FISH. AB - A newly developed device for performing fluorescence in situ hybridization (FISH) is described. An adapted procedure was compared with two typical FISH protocols. Tests were performed with Pseudomonas cells and the gene probe EUB338. With the novel procedure, we obtained a better recovery of cells and less variability in results. PMID- 11943365 TI - Europe and the stem cell debate. PMID- 11943366 TI - Post-genomic applications of lab-on-a-chip and microarrays. PMID- 11943367 TI - Entrapment of proteins and DNA in thermally evaporated lipid films. AB - The immobilization of biomacromolecules such as proteins, enzymes and DNA in various inert matrices is a problem that attracts considerable attention and is motivated by fundamental, biomedical and industrial interests. In addition to several other entrapping matrices, lipids in the form of monolayers and bilayers are versatile hosts owing to their membrane-mimicking capability, bio friendliness, flexibility and inertness. Here, we discuss the immobilization of proteins, enzymes and DNA via electrostatic interactions in films of thermally evaporated fatty lipids. The role of the lipid in preserving the natural conformation of the biomolecule, protection against harsh environmental conditions and accessibility to substrates and reagents is an important feature of the protocol and is highlighted. PMID- 11943372 TI - Bioterrorism: how well are we protected? PMID- 11943373 TI - Advanced agricultural biotechnologies and sustainable agriculture. AB - Agricultural biotechnologies are anchored to a scientific paradigm rooted in experimental biology, whereas sustainable agriculture rests on a biological paradigm that is best described as ecological. Both biotechnology and sustainable agriculture are associated with particular social science paradigms: biotechnology has its foundation in neoclassical economics, but sustainability is framed by an emerging community-centered, problem-solving perspective. Fundamentally, biotechnology and neoclassical economics are reductionist in nature. Sustainability and community problem-solving, however, are nonreductionist. Given these differences, we might see the development of two rather distinct systems of food production in the near future. PMID- 11943374 TI - From patenting genes to proteins: the search for utility via function. AB - The debate regarding the patenting of genes has extended into the post-genome era. With only approximately 35000 genes deduced from the draft sequence of the human genome, there are fears that a few companies have already gained monopoly on the potential benefits from this knowledge. Nevertheless, it is accepted that proteins determine gene function and function is not readily predicted from gene sequence. Furthermore, genes can encode multiple proteins and a single protein can have multiple functions. Here, we argue that unraveling the intrinsic complexity of proteins and their functions is the key towards determining the utility requirement for patenting protein inventions and consider the possible socioeconomic impact. PMID- 11943375 TI - Filamentous fungi as cell factories for heterologous protein production. AB - Filamentous fungi have been used as sources of metabolites and enzymes for centuries. For about two decades, molecular genetic tools have enabled us to use these organisms to express extra copies of both endogenous and exogenous genes. This review of current practice reveals that molecular tools have enabled several new developments. But it has been process development that has driven the final breakthrough to achieving commercially relevant quantities of protein. Recent research into gene expression in filamentous fungi has explored their wealth of genetic diversity with a view to exploiting them as expression hosts and as a source of new genes. Inevitably, the progress in the 'genomics' technology will further develop high-throughput technologies for these organisms. PMID- 11943376 TI - Peptide mimotopes as surrogate antigens of carbohydrates in vaccine discovery. AB - Carbohydrate antigens are immune targets associated with a variety of pathogens and tumor cells. Unfortunately, most carbohydrates are intrinsically T cell independent antigens, which diminishes their efficacy as immunogens. The conversion of carbohydrate epitopes to peptide mimotopes is one means to overcome the T cell-independent nature of carbohydrate antigens because peptides have an absolute requirement for T cells. Although such conversion has great potential for the development of veterinarian and human vaccines, there are issues related to the use of peptide-based immunogens as functional surrogates. Some of these issues are fundamental, pertaining to how mimicry comes about at the molecular level, and some are application oriented, directed at elucidating important immunological mechanisms. In this article the potential and caveats of this technology regarding its application in vaccine discovery are analyzed. PMID- 11943378 TI - Ronald H.A. Plasterk. Interview by Suzanne Berry. PMID- 11943377 TI - Detection of genetically modified organisms in foods. AB - Legislation enacted worldwide to regulate the presence of genetically modified organisms (GMOs) in crops, foods and ingredients, necessitated the development of reliable and sensitive methods for GMO detection. In this article, protein- and DNA-based methods employing western blots, enzyme-linked immunosorbant assay, lateral flow strips, Southern blots, qualitative-, quantitative-, real-time- and limiting dilution-PCR methods, are discussed. Where information on modified gene sequences is not available, new approaches, such as near-infrared spectrometry, might tackle the problem of detection of non-approved genetically modified (GM) foods. The efficiency of screening, identification and confirmation strategies should be examined with respect to false-positive rates, disappearance of marker genes, increased use of specific regulator sequences and the increasing number of GM foods. PMID- 11943383 TI - Oleic acid, a skin penetration enhancer, affects Langerhans cells and corneocytes. AB - Permeation enhancers (PE) are frequently used in the field of dermal research and for the development of transdermal delivery products. However, their influence on skin epidermal Langerhans cells (LC) has not yet been investigated. In this work we studied the effect of four PE, oleic acid (OA), propylene glycol (PG), ethanol, and diethylene glycol monoethyl ether (DGME), and an iontophoretic treatment on the morphometric parameters of epidermal Langerhans cells (LC). Retinoic acid (RA) was used as a positive control. Test solutions were applied to the footpad of Sabra mice. The area, perimeter, density and shape factor (SF) were the morphometric parameters evaluated following ATPase staining of LC. Application of RA led to a large decrease in cell density (-50.2%, P<0.01) and dendritic shape (19.8%, P<0.01). Treatment with 10% OA in ethanolic solution caused a severe decrease in LC density (-69.0%, P<0.01), accompanied by a decrease in dendricity as measured by the changes in SF. Ethanol had no statistically significant effect on the LC morphologic parameters tested. All other PE had a mild, if any, effect on LC morphology. SEM micrographs of the skin of IOPS hairless rats demonstrated that 24 h in vivo treatment with 10% OA in ethanolic solution resulted in the generation of pores on the surface of epidermal corneocytes. PMID- 11943384 TI - Biodegradable injectable in situ forming drug delivery systems. AB - The ability to inject a drug incorporated into a polymer to a localized site and have the polymer form a semi-solid drug depot has a number of advantages. Among these advantages is ease of application and localized, prolonged drug delivery. For these reasons a large number of in situ setting polymeric delivery systems have been developed and investigated for use in delivering a wide variety of drugs. In this article we introduce the various strategies that have been used to prepare in situ setting systems, and outline their advantages and disadvantages as localized drug delivery systems. PMID- 11943385 TI - Loading anticancer drugs into HDL as well as LDL has little affect on properties of complexes and enhances cytotoxicity to human carcinoma cells. AB - Low density lipoprotein (LDL) has been found to represent a suitable carrier for cytotoxic drugs that may target them to cancer. This study investigated whether very low density lipoprotein (VLDL), LDL and high density lipoprotein (HDL) can be used to effectively incorporate four cytotoxic drugs, 5-fluorouracil (5-FU), 5 iododeoxyuridine (IUdR), doxorubicin (Dox) and vindesine; characterized the complexes; and examined the effect of incorporation on drug cytotoxicity against HeLa cervical and MCF-7 breast carcinoma cells. Significant drug loading was achieved into all three classes of lipoproteins, consistent with the sizes and hydrophobicity of the drugs. The relative loading efficiency was found to be vindesine>IUdR>Dox>5-FU for all three classes of lipoproteins. As shown by electron microscopy (EM), drug incorporation did not affect the size or morphology of the lipoproteins. Differential scanning calorimetry (DSC) showed that drug loading did not significantly change the thermal transition temperature of core lipids in the lipoproteins. The transition enthalpy was changed only for LDL-Dox and LDL-vindesine. The drugs remained stable in the lipoproteins as determined by high performance liquid chromatography (HPLC). EM, DSC and HPLC data suggest that drugs were incorporated into lipoproteins without disrupting their integrity and drugs remained in their stable forms inside lipoproteins. Compared with free drugs in cytotoxicity assays, the IC(50) values of LDL- and HDL-drug complexes were significantly lower (2.4- to 8.6-fold for LDL complexes and 2.5- to 23-fold for HDL complexes). All free or lipoprotein-bound drug formulations were comparably more cytotoxic against MCF-7 than HeLa cells. Upregulating the lipoprotein receptors enhanced, and downregulating them inhibited, the cytotoxicity, indicating the mechanistic involvement of lipoprotein receptor pathways. Complexes of all four drugs with VLDL, in contrast to LDL and HDL, had the same cytotoxicity as the four corresponding free drugs. Our results suggest that further studies are required of the potential of HDL to be a cancer targeting drug carrier. PMID- 11943386 TI - Guar-based monolithic matrix systems: effect of ionizable and non-ionizable substances and excipients on gel dynamics and release kinetics. AB - The effect of ionic and non-ionic excipients and additives as modulators of swelling and erosion kinetics and verapamil HCl release from guar-based matrix tablets was investigated. Tablet dissolution, erosion and water uptake studies were carried out using a modified USP 23 Apparatus 2 method. The kinetics of gel strength and texture development were studied by textural analysis. Near linear drug release over 24 h was obtained from formulations containing water soluble, ionizable sodium chloride and glycine. The contribution of Fickian release to overall drug release was lowest for these formulations and was correlated with greater gel strength and lower water uptake in the early time period. For soluble sugars (lactose and sucrose) the Fickian contribution to overall drug release was large and associated with pronounced curvilinear profiles. Water uptake was greatest for these additives (450% in 6 h). The lowest water uptake and negligible matrix erosion was observed for microcrystalline cellulose. Release from this formulation was predominantly Fickian. It was found that the physico chemical nature of added excipients significantly influences the release kinetics from guar-based formulations. Ionic, water soluble materials (sodium chloride, glycine) reduce initial hydration of the matrix and thus have the ability to limit the initial rapid diffusion of drug and to sustain near linear release over 24 h. PMID- 11943387 TI - Time-programmed pulsatile release of dextran from calcium-alginate gel beads coated with carboxy-n-propylacrylamide copolymers. AB - Time-programmed release of macromolecular drugs was achieved by utilization of calcium-alginate gel beads modified with coated copolymer layers. Modified calcium-alginate gel beads coated with poly(carboxy-n-propylacrylamide-co dimethylacrylamide) [poly(CNPAAm-co-DMAAm)] (22.7 mol% of CNPAAm) of varying coating thickness from 25 to 125 microm were developed as drug carriers. Model macromolecular drugs used were fluorescein isothiocyanate (FITC)-labeled dextrans with different molecular weights ranging from 9400 to 145000. FITC-dextran release was strongly dependent on both copolymer coating thicknesses and the dextran molecular weights. Release of FITC-dextran (MW 9400) followed Fickian diffusion according to t(1/2) dependence, indicating that the drug diffusion is the main driving force for release of dextran MW 9400. Release of higher molecular weight FITC-dextrans (71,00 and 145,00) exhibited a burst-effect preceded by a preset lag time. These release profiles were governed by the dissociation of calcium ions from polyguluronate sequences in alginate molecules along with the diffusion of sodium ions into the gel bead core. This created osmotic pressure inside the gel, inducing breakage of the coated copolymer layer and accelerated drug release. Burst release of macromolecular drugs thus occurred after a certain lag period. The lag time was regulated by the copolymer coat thickness. A pulsatile release of FITC-dextran was demonstrated by combining a series of modified alginate gel beads in a single batch. PMID- 11943388 TI - Temperature-responsive and degradable hyaluronic acid/Pluronic composite hydrogels for controlled release of human growth hormone. AB - Temperature-sensitive hyaluronic acid (HA) hydrogels were synthesized by photopolymerization of vinyl group modified HA in combination with acrylate group end-capped poly(ethylene glycol)-poly(propylene glycol)-poly(ethylene glycol) tri block copolymer (Pluronic F127). The synthesized HA/Pluronic composite hydrogels gradually collapsed with increasing temperature over the range of 5-40 degrees C, suggesting that the Pluronic component formed self-associating micelles in the hydrogel structure. Upon prolonged incubation in a buffer medium, the micelles slowly degraded due to the hydrolytic scission of the ester linkage between the Pluronic and acrylate group. The mass erosion occurred much faster at 37 degrees C than at 13 degrees C, indicating that at the higher temperature, the ester linkage between the Pluronic and acrylate group might be more exposed to an aqueous environment and thus be more readily hydrolyzed due to Pluronic micellization. Incorporation of recombinant human growth hormone in the hydrogel resulted in a sustained release profile which followed a mass erosion pattern. PMID- 11943389 TI - Magnetic resonance imaging study of the swelling kinetics of hydroxypropylmethylcellulose (HPMC) in water. AB - Magnetic resonance imaging has been used to monitor the hydration of hydroxypropylmethylcellulose samples by two-dimensional mapping of properties such as spin density and relaxation times. The measurements were performed at two pH values of water: 2 and 6 and two temperatures 25 and 37 degrees C. It is shown that transport behavior of water into HPMC changes from being almost completely relaxation controlled (case II) at pH=2 to Fickian behavior for pH=6. It was also observed that radial swelling is larger for the system composed of HPMC and water at pH=6 than at pH=2. PMID- 11943390 TI - The release of model macromolecules may be controlled by the hydrophobicity of palmitoyl glycol chitosan hydrogels. AB - A non-covalently cross-linked palmitoyl glycol chitosan (GCP) hydrogel has been evaluated as an erodible controlled release system for the delivery of hydrophilic macromolecules. Samples of GCP with hydrophobicity decreasing in the order GCP12>GCP11>GCP21 were synthesised and characterised by 1H NMR. Hydrogels were prepared by freeze-drying an aqueous dispersion of the polymer in the presence or absence of either a model macromolecule fluorescein isothiocyanate dextran (FITC-dextran, MW 4400), and/or amphiphilic derivatives Gelucire 50/13 or vitamin E d-alpha-tocopherol polyethylene glycol succinate. Gels were analysed for aqueous hydration, FITC-dextran release, and bioadhesion, and imaged by scanning electron microscopy. The gels were highly porous and could be hydrated to up to 95x their original weight without an appreciable volume change and most gels eventually eroded. Hydration and erosion were governed by the hydrophobicity of the gel and the presence of the amphiphilic additives. GCP gels could be loaded with up to 27.5% (w/w) of FITC-dextran by freeze-drying a dispersion of GCP in a solution of FITC-dextran. The controlled release of FITC-dextran was governed by the hydrophobicity of the gel following the trend GCP21>GCP11>GCP12. GCP gels were bioadhesive but less so than hydroxypropylmethylcellulose, Carbopol 974NF (7:3) tablets. PMID- 11943391 TI - Differences in the intracellular fate of free and polymer-bound doxorubicin. AB - Internalization and subcellular fate of free doxorubicin or its polymeric conjugates based on poly N-(2-hydroxypropyl)methacrylamide (pHPMA), either non targeted or targeted with anti-Thy1.2 or anti-CD71 monoclonal antibody was tested on EL-4 mouse T-cell lymphoma, SW620 human colorectal carcinoma and OVCAR-3 human ovarian adenocarcinoma. Doxorubicin fluorescence allowed us to follow the internalization and intracellular distribution of tested conjugates by laser scanning confocal microscopy and/or by fluorescent microscopy. Whereas free doxorubicin was always detectable only in the nuclei of treated cells, detectable fluorescence of doxorubicin bound to a polymeric carrier, targeted or non targeted, was detectable up to 3 days of incubation only in the cytoplasmatic structures. While free doxorubicin causes apoptosis in the populations of tested cancer cell lines, significant number of apoptotic cells was never found in cell cultures exposed to targeted or non-targeted polymeric conjugates. In contrast to free doxorubicin, which is a strong inducer of p53 expression, increased p53 expression was never observed after the treatment with the polymeric drug. High performance liquid chromatographic analysis shows that the percentage of cleaved doxorubicin is very low even after 48 h of incubation of tested cells with the polymeric conjugate, and cannot be the only reason for the toxicity of the conjugate. We suggest that: (a) after the treatment with pHPMA-bound drug, the cells die by necrosis and (b) the toxicity of pHPMA-based conjugates is a combination of the toxic effect of released doxorubicin and the toxic effect of doxorubicin in polymer-bound form directed against cell membranes. PMID- 11943392 TI - In vitro evaluation of a system for pH-controlled peroral delivery of metformin. AB - Oral absorption of the antihyperglycaemic agent metformin (MF x HCl) is confined to the upper part of the intestine, therefore controlled-release oral formulations of this drug should ensure a complete release during transit from stomach to jejunum. Compressed matrix tablets based on pH-sensitive poly(ethylene oxide) (PEO)-Eudragit L100 (EUD L) compounds have shown in vitro a compliance with the above requirement. The polymer compounds were prepared by a coevaporation process. The release pattern of MF x HCl from matrices depended on the PEO-EUD L ratio in the coevaporate. The 1:1 (w/w) ratio was unable to control MF x HCl release in simulated gastric fluid (SGF, pH 1.2), because the matrix material was excessively hydrophilic. Nevertheless, the release rate in SGF could be modulated by increasing the EUD L fraction in the coevaporate. With a PEO (M(w), 400 kDa)-EUD L (1:2, w/w) ratio the percent dose released in 2 h to SGF, where the coevaporate was insoluble, was around 23 or 50% with 10 or 20% loading dose. The release was then completed within the successive 2 h of elution with simulated jejunal fluid (SJF, pH 6.8) where EUD L and the coevaporate gradually dissolved. Release in SGF was controlled by matrix swelling and/or drug diffusion in matrix, whereas matrix dissolution controlled release in SJF. The unique release-controlling properties of the polymer compounds were due to PEO-EUD L macromolecular interactions. Matrices show promise of a gradual and complete release of MF x HCl from stomach to jejunum, unaffected by gastric pH fluctuations. This mode of administration might allow the use of lower therapeutic doses compared to existing immediate- or sustained-release products, thus minimising side effects. PMID- 11943394 TI - pH-induced solubility transition of sulfonamide-based polymers. AB - As an approach to designing new pH-sensitive polymers for bio-related application, we have modified selected sulfonamides, with various pK(a), to polymerizable monomers. The pK(a) of the monomers, homopolymers, and copolymers with N,N-dimethylacrylamide were examined, and pH-induced phase transition behavior, particularly in solubility, was investigated. The pK(a) of sulfonamide monomers and polymers at 25 degrees C was slightly higher than those of corresponding sulfonamides but the enthalpy of the ionization was influenced due to interfering resonance structures that are present in the mother compounds. The solubility transition of each homopolymer in aqueous solutions occurred at a degree of ionization of 85-90%. For the copolymers, the solubility transition observed by light transmittance completed in a narrow pH range (0.2-0.3 pH units) and this transition pH shifted to a higher pH region with the increasing sulfonamide unit in the copolymer, though the pK(a) was not considerably changed. This polymer precipitation occurs because of the relative balance of overall hydrophilicity/hydrophobicity along the polymer chain. Because the unionized form of sulfonamide units is considered to be hydrophobic, the copolymer with higher content of sulfonamide unit requires a higher degree of ionization for solubilization and thus solubility transition occurred at higher pH. PMID- 11943393 TI - Vitamin E TPGS used as emulsifier in the solvent evaporation/extraction technique for fabrication of polymeric nanospheres for controlled release of paclitaxel (Taxol). AB - The D-alpha-tocopheryl polyethylene glycol 1000 succinate (vitamin E TPGS) was applied in the present investigation as surfactant stabiliser to fabricate paclitaxel-loaded PLGA nanospheres in the solvent evaporation/extraction technique with successful achievement. Laser light scattering system (LLS), scanning electron microscopy (SEM), atomic force microscopy (AFM), differential scanning calorimetry (DSC), Fourier transform infra-red spectroscopy (FTIR), and X-ray photoelectron spectroscopy (XPS) were employed to characterise the nanopsheres fabricated in various recipes under various preparation conditions for size and size distribution, surface morphology, thermogram property and surface chemistry. Encapsulation efficiency and in vitro release was measured by the high-performance liquid chromatography (HPLC). The outcomes were discussed with respect to the development of polymeric nanospheres delivery system of the anticancer drug, paclitaxel (Taxol((R))). The produced nanospheres were found in fine spherical shape with smooth surfaces and without aggregation or adhesion. There was no significant difference in morphology between the vitamin E TPGS emulsified and PVA emulsified PLGA nanospheres. However, it was found that, in comparison with the traditional chemical emulsifier PVA, the TPGS could significantly improve the encapsulation efficiency of the drug in the PLGA nanospheres, which could be as high as 100%. The size of the vitamin E TPGS emulsified nanospheres ranged from 300 to 800 nm and the size distribution was narrow with polydispersity of 0.005-0.045. XPS investigation demonstrated that there were residual surfactant molecules remained on the surface although the TPGS could be washed out relatively thoroughly in the process of nanospheres formation. This finding was also confirmed by FTIR-PAS investigation of the nanospheres. The in vitro release indicated that the release property of paclitaxel from the nanospheres strongly depends on the emulsifier type employed in the fabrication. Our research shows that vitamin E TPGS could be an ideal and effective emulsifier. PMID- 11943395 TI - The effect of poly(ethylene glycol)-poly(D,L-lactic acid) diblock copolymers on peptide acylation. AB - The combination of poly(ethylene glycol) (PEG) with a biodegradable poly(ester), such as poly(D,L-lactic acid) (PLA), is an approach that has been successfully used for the stabilization of proteins and peptides in several biodegradable delivery devices. The acylation of peptides inside degrading PLA microspheres has been described only recently as another instability mechanism related to the accumulation of polymer degradation products inside eroding PLA. We investigated whether the block copolymerization of PLA with PEG reduces peptide acylation inside degrading microspheres. Diblock copolymers consisting of poly(D,L-lactic acid) covalently bound to poly(ethylene glycol)-monomethyl ether (Me.PEG-PLA) were used for these investigations. Human atrial natriuretic peptide (ANP) was incorporated into microspheres manufactured from Me.PEG5-PLA45, a diblock copolymer with an overall PEG content of 10%. Peptide integrity inside the microspheres was monitored by HPLC-MS analysis during 4 weeks of microsphere degradation in isotonic phosphate buffer (pH 7.4) at 37 degrees C. Inside the degrading Me.PEG5-PLA45 microspheres, acylation products as well as an oxidation product of ANP were formed. The results demonstrate that the combination of PEG with PLA does not necessarily display a favorable effect concerning peptide acylation inside degrading polymer microspheres. However, they also suggested that the acylation reaction is mainly driven by the formation and accumulation of polymer degradation products inside the degrading microspheres. PMID- 11943396 TI - Modulated insulin permeation across a glucose-sensitive polymeric composite membrane. AB - A glucose-sensitive polymeric composite membrane was prepared based on our previously developed stimuli-responsive membrane system. Membranes were cast from a mixture of glucose oxidase (GOD), catalase, and poly(N-isopropylacrylamide-co methacrylic acid) (poly(NIPAm/MAA)) nanoparticles dispersed in a solution of a hydrophobic polymer. High efficiency of enzyme immobilization was achieved with undetectable leakage. The bioactivity of the immobilized GOD, as measured by pH change of glucose solutions, was found to be equivalent to approximately 80% of that of the free GOD. The addition of catalase markedly increased the oxidation rate of glucose. However, an optimal unit ratio of GOD to catalase and optimal enzyme loading were observed. The rate of insulin permeation through the membrane was modulated by glucose concentration due to shrinking or swelling of the embedded pH-sensitive nanoparticles. The response of insulin permeability to the change in the glucose concentration could be detected within 5-15 min. The permeability of insulin increased more than 3-fold as the glucose concentration was raised from 50 to 200 mg/dl. The average insulin permeability at 400 mg/dl of glucose was 8-fold that at 50 mg/dl in a continuous test in saline and was 6-fold in a three-cycle discontinuous test in pH 7.4 buffer. PMID- 11943397 TI - Immobilized liposome layers for drug delivery applications: inhibition of angiogenesis. AB - Liposomes were immobilized onto the surface of perfluorinated polymer tape samples and tissue culture polystyrene well-plates using a multilayer immobilization strategy. In the first step, a thin interfacial bonding layer with surface aldehyde groups was deposited from a glow discharge struck in acetaldehyde vapour. Polyethylenimine was then covalently bound onto the aldehyde groups by reductive amination, followed by covalent binding of NHS-PEG-biotin molecules onto the surface amine groups by carbodiimide chemistry. Next, NeutrAvidin protein molecules were bound onto the PEG-biotin layer. Finally, liposomes containing PEG-biotinylated lipids were docked onto the remaining binding sites of the surface-immobilized NeutrAvidin molecules. AFM was used to image surface-bound liposomes and revealed a density well below close packing. The release characteristics of the surface-bound liposomes were measured by the fluorescence intensity changes of carboxyfluorescein upon release. Liposomes filled with sodium orthovanadate were surface immobilized and used in two in vitro angiogenesis assays. Marked differences compared to various control samples were observed, demonstrating the utility of drug-filled, surface-bound liposomes for evoking localized, controlled biological host responses proximal to an implanted biomedical device. PMID- 11943398 TI - Physicochemical behavior and cytotoxic effects of p(methacrylic acid-g-ethylene glycol) nanospheres for oral delivery of proteins. AB - The challenges faced to orally deliver therapeutic agents with unfavorable physicochemical properties, such as proteins, have been the primary motivation for the design and development of novel oral delivery systems that could circumvent biological barriers. In this work, we examined complexation-sensitive hydrogel nanospheres composed of poly[methacrylic acid-grafted-poly(ethylene glycol)] (P(MAA-g-EG)), on a model biological environment. For this purpose, a gastrointestinal cell culture model, the Caco-2 cell line, was employed to investigate the cytotoxic effects of the polymeric carrier and its effects on the cell monolayer integrity. The determination of the cytotoxic effects of the polymer network on the cell monolayer was performed by a colorimetric assay and by the counting of viable cells using the trypan blue exclusion method. Electrophysiological measurements were performed to measure the transepithelial electrical resistance changes in the monolayers in the presence and absence of the nanosphere suspension. The examination of the physicochemical interactions of the P(MAA-g-EG) nanosphere system with Caco-2 cell monolayers revealed that these systems possessed low cytotoxicity and were capable of opening the tight junctions between epithelial cells, therefore significantly reducing the transepithelial electrical resistance. PMID- 11943399 TI - Mucoadhesive beclomethasone microspheres for powder inhalation: their pharmacokinetics and pharmacodynamics evaluation. AB - The feasibility of prolonging drug action and/or reducing drug dosage using mucoadhesive beclomethasone dipropionate (BDP) microspheres for powder inhalation was investigated. BDP was spray-dried from ethanol solution or aqueous suspension systems dissolving a mucoadhesive polymer, hydroxypropylcellulose (HPC); this resulted in amorphous and crystalline BDP incorporation in the HPC microspheres (aBDP/HPC and cBDP/HPC; BDP-HPC ratio=1:4), respectively. These microspheres were administered as powder aerosols to healthy or antigen-induced, asthmatic guinea pigs, and BDP's retention in the lung (pharmacokinetics) and inhibitory duration with respect to eosinophil infiltration into the airways (pharmacodynamics) were compared to those for pure crystalline BDP (cBDP; 'control'). Both BDP/HPC microspheres were prepared within a respirable-size range of 2.5-2.9 microm. BDP's aqueous solubility was increased 25 times for aBDP/HPC, compared to crystalline counterpart. Pharmacokinetic profiles for three powders were dissolution-modulated. aBDP/HPC showed rapid BDP absorption from the lung (> or = 95% absorption for 180 min) with a greater metabolite (B17MP) formation, compared to cBDP, primarily due to the increased dissolution of amorphous BDP. In contrast, 86.0% of BDP remained at 180 min following cBDP/HPC administration, demonstrating the prolonged BDP's retention in the lung by virtue of poor dissolution (and/or release) and retarded mucociliary clearance. As a result, while cBDP (1.37 mg/kg) significantly inhibited eosinophil infiltration into the lungs of antigen-sensitized and -challenged guinea pigs for only 1-6 h, cBDP/HPC, despite a much lower drug dosage (0.25 mg/kg), was capable of maintaining such inhibitory effects for 24 h following administration. It appeared therefore that the prolonged lung retention of BDP by the use of the HPC microspheres (cBDP/HPC) was attributed to prolonging its pharmacological duration without requiring increased drug dosage. PMID- 11943400 TI - Multivalent interactions between biotin-polyrotaxane conjugates and streptavidin as a model of new targeting for transporters. AB - Kinetic analysis of interactions between biotin-polyrotaxane or biotin-alpha cyclodextrin (biotin-alpha-CD) conjugates and streptavidin was carried out as a model of new targeting to transporters using the surface plasmon resonance (SPR) technique. The biotin-polyrotaxane conjugates, in which biotin-introduced alpha CDs are threaded onto a poly(ethylene oxide) chain capped with bulky end-groups, are expected to increase the valency of biotin from monovalent to multivalent binding. The number of biotins conjugated with one polyrotaxane molecule varied from 11 to 78, and apparently increased the association equilibrium constant (K(a)), assuming pseudo-first-order kinetics. A detailed dissociation kinetics was analyzed and the re-binding of the biotin-polyrotaxane conjugates was observed on the streptavidin-deposited SPR surface. The magnitude of the re binding is likely to become larger with increasing the number of biotins, suggesting multivalent interaction on the SPR surface. To quantify the effect of valency, competitive inhibition assay was performed in terms of the supramolecular structure of the polyrotaxane. The inhibitory potency of the biotin-polyrotaxane conjugate was found to be 4-5 times greater than that of the biotin-alpha-CD conjugate. Therefore, the biotin-polyrotaxane conjugates by supramolecular formation of the biotin-alpha-CD conjugate significantly switches from monovalent to multivalent bindings to the model binding protein, streptavidin. PMID- 11943401 TI - Simulation and parametric study of a film-coated controlled-release pharmaceutical. AB - Pharmaceutical formulations can be designed as Multiple Unit Systems, such as Roxiam CR, studied in this work. The dose is administrated as a capsule, which contains about 100 individual pellets, which in turn contain the active drug remoxipride. Experimental data for a large number of single pellets can be obtained by studying the release using microtitre plates. This makes it possible to study the release of the individual subunits making up the total dose. A mathematical model for simulating the release of remoxipride from single film coated pellets is presented including internal and external mass transfer hindrance apart from the most important film resistance. The model can successfully simulate the release of remoxipride from single film-coated pellets if the lag phase of the experimental data is ignored. This was shown to have a minor influence on the release rate. The use of the present model is demonstrated by a parametric study showing that the release process is film-controlled, i.e. is limited by the mass transport through the polymer coating. The model was used to fit the film thickness and the drug loading to the experimental release data. The variation in the fitted values was similar to that obtained in the experiments. PMID- 11943402 TI - Reversible adsorption by a pH- and temperature-sensitive acrylic hydrogel. AB - Thermo- and pH-sensitive hydrogels were synthesized using N-isopropylacrylamide (NIPA) and N-aminopropylmethacrylamide, cross-linked with N,N' methylenebis(acrylamide). The dependence of the degree of swelling on the cross linking density was analyzed according to the Flory-Huggins theory and a master curve obtained. To optimize the efficiency of these hydrogels in controlled release, we studied the loading and release of a divalent molecule (naphthalenedisulfonic acid, NS-2) in media of different ionic strengths and pH. The uptake process followed the Langmuir adsorption isotherm model. The highest loading occurred when the amino groups in the gel were protonated (acidic pH) and could come close each other to form a binding site for the two sulfonic groups of NS-2, i.e. low degree of cross-linking and collapsed state. Below the phase transition temperature (33 degrees C), NS-2 loaded hydrogels quickly released a significant amount of adsorbate until a new equilibrium between free NS-2 and adsorbed NS-2 was achieved. Above that temperature, hydrogels not only stopped the release but were even able to take free NS-2 up again from the medium, showing that the loading/release process was reversible and reproducible after several temperature cycles. At 37 degrees C, the release rate was independent of the degree of cross-linking (NIPA caused the hydrogel to collapse), but was strongly affected by the pH and salt concentrations of the medium, which condition the strength of the interaction between the hydrogel amino groups and the NS-2 sulfonic groups. In an acidic medium, the protonated amino groups bind NS-2 strongly and the amount released is small. In contrast, at pH 7.4 or in the presence of a high salt concentration, the hydrogel loses its affinity for NS-2 and the release rate increases, giving pH- or salt-sensitive delivery systems. Additionally, since the hydrogel is collapsed, the release can be prolonged for a long period of time. PMID- 11943403 TI - Poly(ethylene glycol) (PEG) conjugated arginine deiminase: effects of PEG formulations on its pharmacological properties. AB - Some tumors, such as melanomas and hepatocellular carcinomas, have a unique nutritional requirement for arginine. Thus, enzymatic degradation of extracellular arginine is one possible means for inhibiting these tumors. Arginine deiminase is an arginine degrading enzyme (ADI) that has been studied as an anti-cancer enzyme. However, ADI has a short serum half-life and, as a microbial enzyme, is highly immunogenic. Formulation of other therapeutic proteins with poly(ethylene glycol) (PEG) has overcome these problems. Here, ADI PEGs were synthesized using PEGs of varying size, structure (linear or branched chain) and linker chemistries. All ADI-PEGs retained approximately 50% of enzyme activity when PEG was covalently attached to approximately 40% of the primary amines irrespective of the PEG molecular weight or attachment chemistry used. However, it was observed that, as the PEG size increases to 20 kDa, there was a corresponding increase in the pharmacokinetic (pK) and pharmacodynamic (pD) properties of the formulation. Variation in PEG linker or structure, or the use of PEGs >20,000 mw, did not affect the pK or pD. As has been shown with other therapeutic proteins, repeated injection of ADI-PEG into experimental animals resulted in significantly lower titers of antibodies against this protein than unmodified ADI. These data suggest that formulation of ADI with PEG of 20,000 mw results is the optimal method for formulating this promising therapeutic agent. PMID- 11943404 TI - Biodegradable poly(ethylenimine) for plasmid DNA delivery. AB - Poly(ethylenimine) (PEI) has been known as an efficient gene carrier with the highest cationic charge potential. High transfection efficiency of PEI, along with its cytotoxicity, strongly depends on the molecular weight. Synthesis of cationic copolymers derived from the low molecular weight of PEI and hydrophilic poly(ethylene glycol) (PEG), which are water soluble and degradable under physiological conditions, was investigated for plasmid delivery. Hydrophilic PEG is expected to reduce the toxicity of the copolymer, improve the poor solubility of the PEI and DNA complexes, and help to introduce degradable bonds by reaction with the primary amines in the PEI. Considering the dependence of transfection efficiency and cytotoxicity on the molecular weight of the PEI, high transfection efficiency is expected from an increased molecular weight of the copolymer and low cytotoxicity from the introduction of PEG and the degradation of the copolymer into low molecular weight PEIs. Reaction conditions were carefully controlled to produce water soluble copolymers. Results from a gel retardation assay and zetapotentiometer indicated that complete neutralization of the complexes was achieved at the charge ratios of copolymer/pSV-beta-gal plasmid from 0.8 to 1.0 with the mean particle size of the polyplexes ranging from 129.8+/-0.9 to 151.8+/-3.4 nm. In vitro transfection efficiency of the synthesized copolymer increased up to three times higher than that of starting low molecular weight PEI, while the cell viability was maintained over 80%. PMID- 11943405 TI - Inhibition of liver metastasis by targeting of immunomodulators using mannosylated liposome carriers. AB - Mannosylated liposomes were prepared by incorporating cholesten-5-yloxy-N-(4-((1 imino-2-beta-D-thiomannosylethyl)amino)butyl)formamide (Man-C4-Chol) into small unilamellar liposomes consisting of cholesterol and distearoyl phosphatidylcholine (DSPC). The biodistribution of liposomes labeled with [3H]cholesteryl hexadecyl ether was examined in mice. The rate and extent of the hepatic uptake of those [3H]liposomes increased proportionally on increasing the mixing ratio of Man-C4-Chol. Their hepatic uptake was reduced by increasing the administered dose due to the limited number of mannose receptors. The liver uptake of [3H]Man-liposomes was preferentially mediated by liver non-parenchymal cells (NPC) and significantly inhibited by co-injection with an excess of Man BSA, indicating the involvement of a mannose receptor-mediated mechanism in the hepatic uptake of Man-liposomes. Muramyl dipeptide (MDP), an immunomodulator, was also incorporated into the liposomes and its inhibitory effect in an experimental liver metastasis model was examined. In contrast to free MDP treatment, which showed little effect on the inhibition of metastasis, liposomal MDP significantly reduced the number of metastatic colonies in the liver. Active targeting of MDP to liver NPC by Man-liposomes resulted in more effective inhibition than delivery of MDP by liposomes without mannose. Treatment with MDP/Man-liposomes further increased the survival of the tumor-bearing mice. These results suggest that Man liposomes are effective carriers for targeted delivery of bioactive compounds to liver NPC. PMID- 11943406 TI - Anti-tumor effect of intraperitoneal administration of cisplatin-loaded microspheres to human tumor xenografted nude mice. AB - This study evaluates the anti-tumor effect of cisplatin-loaded microspheres (CDDP MS) against peritoneal carcinomatosis using human tumor xenografts. The incorporated CDDP was released from CDDP-MS for 3 weeks in vivo as well as in vitro. CDDP-MS at a dose of 35 mg/kg (at maximal tolerable dose (MTD)) showed effective anti-tumor activity (tumor growth inhibition rate (IR)=70.3%) against Li-7 (human liver cancer) xenografts transplanted into the peritoneal cavity. This procedure also resulted in increased life span (ILS (%)=47.2%), whereas CDDP dissolved in saline solution (CDDP-SOL) at a dose of 8 mg/kg (at MTD) was ineffective (IR=15.7%, ILS=2.6%). Likewise, CDDP-MS (35 mg/kg) significantly prolonged the mean survival time (ILS=50.8%) compared with a CDDP-SOL group (8 mg/kg) (ILS=13.1%) in the mice with Li-7 xenografts transplanted into the spleen. Furthermore, CDDP-MS showed markedly effective anti-tumor activity (IR=82.2%) against H-154 (human stomach cancer) xenografts, in which CDDP-SOL was effective (IR=69.5%) at the MTDs. The suppressive effect of CDDP-MS on accumulation of malignant ascites was intimately related to unchanged CDDP concentration in ascites. These results demonstrated that the administration of CDDP-MS resulted in an unchanged CDDP concentration in ascites, and induced a sustained tumor growth inhibition along with a prolonged survival time. PMID- 11943407 TI - Efficient intracellular drug and gene delivery using folate receptor-targeted pH sensitive liposomes composed of cationic/anionic lipid combinations. AB - pH-sensitive liposomes are designed to promote efficient release of entrapped agents in response to low pH. In this study, novel pH-sensitive liposomes consisting of cationic/anionic lipid combinations are evaluated for intracellular drug and gene delivery. First, liposomes composed of egg phosphatidylcholine, dimethyldioctadecylammonium bromide (DDAB), cholesteryl hemisuccinate (CHEMS), and Tween-80 (25:25:49:1, mol/mol) were shown to stably entrap calcein at pH 7.4 and undergo rapid content release and irreversible aggregation under acidic pH. Compared to pH-sensitive liposomes incorporating dioleoylphosphatidylethanolamine, these liposomes showed improved retention of pH sensitivity in the presence of serum. The folate receptor (FR), which is amplified in a wide variety of human tumors, could be targeted by incorporating 0.1 mol% folate-polyethyleneglycol-phosphatidylethanolamine (f-PEG-PE) into liposomes. f-PEG-PE has been shown to facilitate FR-mediated endocytosis of liposomes into KB human oral cancer cells, which express amplified FR. FR targeted pH-sensitive liposomes produced increased cytosolic release of entrapped calcein, as shown by fluorescence microscopy, and enhanced cytotoxicity of entrapped cytosine-beta-D-arabinofuranoside, as shown by an 11-fold reduction in the IC(50) in KB cells, compared to FR-targeted non-pH-sensitive liposomes. Furthermore, FR-targeted pH-sensitive liposomes composed of DDAB/CHEMS/f-PEG-PE, combined with polylysine-condensed plasmid DNA, were shown to mediate FR-specific delivery of a luciferase reporter gene into KB cells in the presence of 10% serum. These findings suggest that cationic lipid-containing pH-sensitive liposomes, combined with FR targeting, are effective vehicles for intracellular drug and gene delivery. PMID- 11943408 TI - Effect of poly(ethylene glycol) grafts on temperature-sensitivity of thermosensitive polymer-modified liposomes. AB - A copolymer of N-isopropylacrylamide and N-acryloylpyrrolidine, which exhibits a lower critical solution temperature (LCST) at 38 degrees C, having a didodecyl group at the chain terminal was synthesized and calcein-loaded liposomes were prepared from a mixture of dioleoylphosphatidylethanolamine and the copolymer. While the contents release from the copolymer-modified liposomes was strongly accelerated around the LCST of the copolymer, some extent of the release was also observed below the LCST, probably because the partly-dehydrated copolymer chains interacted with the liposome membrane. When poly(ethylene glycol) with the number average molecular weight of 550 having a didodecyl group at the chain end was introduced into the copolymer-modified liposomes, the contents release from the copolymer-modified liposomes was effectively suppressed below the LCST but hardly affected above the LCST. As a result, the liposomes exhibited much sharper enhancement of the contents release in a narrow temperature region. The temperature-sensitive contents release property of the copolymer-modified liposomes became less significant in the presence of serum. However, inclusion of the anchor-bearing poly(ethylene glycol) derivative improved the contents release behavior of the copolymer-modified liposomes in the presence of serum. PMID- 11943409 TI - Controlled release of plasmid DNA from cationized gelatin hydrogels based on hydrogel degradation. AB - This paper shows achievement of the in vivo controlled release of a plasmid DNA from a biodegradable hydrogel and the consequent regulation of gene expression period. Cationization of gelatin was preformed through introduction of ethylenediamine and the gelatin prepared was crosslinked by various concentrations of glutaraldehyde to obtain cationized gelatin (CG) hydrogels as the carrier of plasmid DNA. In vivo release of plasmid DNA from the CG hydrogels was compared with the in vivo degradation of hydrogels. When CG hydrogels incorporating 125I-labeled plasmid DNA were implanted into the femoral muscle of mice, the plasmid DNA radioactivity remaining decreased with time and the retention period prolonged with a decrease in the water content of hydrogels used. The higher the water content of 125I-labeled CG hydrogels, the faster the hydrogel radioactivity remaining decreased with time. The time profile of plasmid DNA remaining in the hydrogels was in good accordance with that of hydrogel radioactivity, irrespective of the water content. Intramuscular implantation of plasmid DNA-incorporated CG hydrogels enhanced significantly expression of the plasmid DNA around the implanted site. The retention period of gene expression became longer as the hydrogel water content decreased. Fluorescent microscopic study revealed that the plasmid DNA-CG complex was detected around the hydrogel implanted even after 7-day implantation in marked contrast to the injection of plasmid DNA solution. It was concluded that in our hydrogel system, active plasmid DNA was released accompanied with the in vivo degradation of hydrogel, resulting in extended gene expression. The time profile of plasmid DNA release and the consequent gene expression was controllable by changing the water content of hydrogels. PMID- 11943410 TI - Enhanced expression of plasmid DNA-cationized gelatin complex by ultrasound in murine muscle. AB - This study is an investigation to experimentally confirm that ultrasound (US) irradiation is effective in enhancing the gene expression of plasmid DNA. A cationized gelatin was prepared by introducing primary amino groups into gelatin. The plasmid of the LacZ gene was complexed with the cationized gelatin and injected into the femoral muscle of normal mice. Following US irradiation at the injected site of muscle, the gene expression of the treated muscle was evaluated to compare with that of the complex injection plus no US irradiation. US irradiation enabled the complex to significantly enhance the gene expression at the injected muscle, in contrast to naked plasmid DNA, although the extent depended on the experimental conditions, such as the injection dose of plasmid DNA, and the time period or timing of US irradiation. The gene-expressed area at the muscle was not changed by the time interval between the complex injection and US irradiation. Fluorescent microscopic observation revealed that the complex was homogeneously distributed in the muscle tissue around the injected site by US irradiation without any dissociation of plasmid DNA. The fluorescent image of plasmid DNA superposed on that of myosin heavy chain indicated intracellular localization of plasmid DNA. We demonstrated that US irradiation is a promising technique to enhance gene expression even in the normal muscle. PMID- 11943411 TI - Preparation of stable insulin-loaded nanospheres of poly(ethylene glycol) macromers and N-isopropyl acrylamide. AB - A series of nanospheres composed of temperature-sensitive poly(N isopropylacrylamide), poly(ethylene glycol) 400 dimethacrylate, and poly(ethylene glycol) 1000 methacrylate was prepared by a thermally-initiated free radical dispersion polymerization method. Insulin was loaded into the nanoparticles by equilibrium partitioning. The loading capacity of insulin into the nanoparticles was 2.1% (2.1 mg insulin/100 mg nanoparticles). The stability of the loaded insulin at elevated temperatures was investigated by reverse phase high pressure liquid chromatography. The nanoparticles were able to protect the loaded insulin, as more than 80% of the loaded insulin could still be detected compared to 0% for the control (0.1% insulin solution in PBS) when heated to 80 degrees C for 5 h. The stability of the loaded insulin at high shear stress (289 1/s) was also investigated. No significant loss of insulin was detected both from nanoparticles loaded with insulin sample and the control (0.1% insulin solution in PBS). The results showed that shear stress alone did not have a major effect on insulin denaturation. The ability of the nanoparticles to protect the insulin from high temperature and high shear stress made the system a good candidate as a carrier for insulin for fluidized bed coating technology. PMID- 11943414 TI - Simvastatin in primary biliary cirrhosis: effects on serum lipids and distinct disease markers. AB - BACKGROUND/AIMS: Primary biliary cirrhosis (PBC) is an autoimmune disease of the liver with inflammation of small and middle-sized bile ducts. Serum lipids are frequently elevated, but the use of a lipid lowering drug therapy in PBC is still a matter of debate. Application of an inhibitor of 3-hydroxy-3-methylglutaryl coenzyme A (HMG-CoA) reductase in hypercholesterolemic PBC patients was therefore the subject of the present study. METHODS: Six female patients (aged 46.5 (32-61) years; median (range)) were treated with the HMG-CoA reductase inhibitor simvastatin (5 or 20 mg/day). Levels of serum total cholesterol, low-density lipoprotein (LDL) cholesterol and high-density lipoprotein (HDL) cholesterol were determined prior to and after 2 months of treatment. Concentrations of serum markers of cholestasis, antimitochondrial antibodies (AMA), and immunoglobulins A, G and M were also assessed. RESULTS: Simvastatin significantly (P<0.05) reduced serum levels of total cholesterol, LDL cholesterol, alkaline phosphatase, -glutamyltransferase, and immunoglobulin M (by 19, 26, 12, 37 and 14%, respectively). CONCLUSIONS: The lipid lowering potency of the HMG-CoA reductase inhibitor simvastatin was confirmed in hypercholesterolemic patients with PBC. The drug might also prove useful as modulator of cholestasis and of immune response in this disease. PMID- 11943415 TI - Lentiviral vectors for efficient transduction of isolated primary quiescent hepatocytes. AB - BACKGROUND/AIMS: Lentiviral vectors were designed to obtain efficient transduction of primary quiescent hepatocytes. METHODS: A hepatitis B virus (HBV) fragment containing enhancers and posttranscriptional regulatory element was used to increase expression levels. The human immunodeficiency virus (HIV) central polypurine tract (PPT) was used to increase transduction of quiescent cells. HBV elements were incorporated downstream and the HIV PPT was incorporated upstream of green fluorescent protein expression cassettes in third generation self inactivating lentiviral vectors. RESULTS: The HBV fragment increased mean fluorescence of transduced HepG2 hepatoma cells 4.3+/-1.7-fold and 2.3-6.0-fold in various other cell types. A role of HBV x protein in the function of the HBV element was excluded. The HBV element increased the number of transducing units per pg of HIV p24 twofold. The unmodified lentiviral vector transduced 5+/-1% of cultured quiescent primary rat hepatocytes, HBV elements increased transduction to 54+/-13% and increased fluorescence 2.8+/-0.6-fold. The PPT increased transduction to 47+/-11% and increased fluorescence 2.3+/-0.4-fold. The vector with PPT and HBV elements transduced 68+/-10% of hepatocytes and increased fluorescence synergistically, 17+/-6 fold. CONCLUSIONS: This study shows that HBV elements or HIV PPT are required for efficient transduction of primary hepatocytes. PMID- 11943416 TI - Inhibition of Kupffer cell activity induces hepatic triglyceride synthesis in fasted rats, independent of lipopolysaccharide challenge. AB - BACKGROUND: Lipopolysaccharides (LPS), cleared from the blood by Kupffer cells, induce hypertriglyceridemia. AIMS: To test the hypothesis that GdCl(3), through inhibition of large Kupffer cell activity, modulates LPS-induced hyperlipidemia in rats. METHODS: Male Wistar rats received a single intravenous injection of GdCl(3)(10 mg/kg) or saline, 24 h before intraperitoneal LPS (1.5 mg/kg) administration. Serum and hepatic lipids as well as activity of key enzymes controlling fatty acid synthesis and esterification in liver tissue were measured. The incorporation of labeled precursors into lipids was assessed in cultured precision-cut liver slices. RESULTS: GdCl(3) does not prevent hypertriglyceridemia occurring in LPS-treated rats. Surprisingly, GdCl(3) per se is able to promote triglycerides accumulation in the liver tissue, an effect related to an increase in hepatic fatty acid esterification. Such an effect also occurs in rats receiving a dietary supplementation with glycine (5%) known to inhibit Kupffer cell secretory capacity. CONCLUSIONS: Large Kupffer cell inhibition does not prevent LPS-induced hypertriglyceridemia and even leads to a metabolic shift of fatty acids towards their esterification and accumulation in the liver tissue, suggesting that Kupffer cells play a role in the regulation of lipid metabolism of the adjacent hepatocytes, independent of any inflammatory stimulus. PMID- 11943417 TI - Frequency of the S65C mutation of HFE and iron overload in 309 subjects heterozygous for C282Y. AB - BACKGROUND/AIMS: HFE-related haemochromatosis is a common disorder of iron metabolism. Most affected individuals are homozygous for the C282Y mutation of HFE. Some are compound heterozygotes for C282Y/H63D. A small proportion have neither of these genotypes. We have investigated the phenotype of compound heterozygotes for C282Y and another missense mutation S65C. METHODS: Genotype for the S65C mutation was determined in 309 subjects heterozygous for C282Y and negative for H63D, referred because of increased serum iron indices or family screening. A control sample comprising 315 individuals was also studied. RESULTS: Twelve individuals were compound heterozygotes for C282Y and S65C. Seven, referred for family screening, had normal serum iron indices. Five subjects had elevated serum iron indices; three of these had elevated hepatic iron and have received treatment for iron overload. Transferrin saturation was significantly elevated in C282Y/S65C compound heterozygotes compared with simple C282Y heterozygotes. CONCLUSIONS: Some C282Y/S65C compound heterozygotes have elevated serum iron indices and iron overload. The penetrance of this genotype is low and other genetic and environmental factors may influence the expression of iron loading. Screening for S65C may be useful in individuals with iron overload who are not homozygous for C282Y or compound heterozygous for C282Y/H63D. PMID- 11943418 TI - Corticosteroids improve short-term survival in patients with severe alcoholic hepatitis (AH): individual data analysis of the last three randomized placebo controlled double blind trials of corticosteroids in severe AH. AB - BACKGROUND/AIMS: Controversy surrounding the efficacy of corticosteroids in severe alcoholic hepatitis (AH) persists. THE AIMS OF OUR STUDY WERE: (a) to analyze individual data of patients with severe AH discriminant function (DF)> or =32 from the last three randomized controlled trials; and (b) to identify the independent prognostic factors associated with short-term survival. METHODS: Individual data were collected from the three principal investigators. Survival analysis was performed at 28 days using the Kaplan-Meier method and log-rank test. The independent prognostic values were assessed by the proportional hazards regression model. RESULTS: About 102 placebo and 113 corticosteroid patients with DF > or =32 were analyzed. At 28 days, corticosteroid patients had significantly higher survival: 84.6+/-3.4% vs. 65.1+/-4.8%, P=0.001. In univariate analysis, corticosteroid treatment, age, DF, albumin, creatinine and encephalopathy were prognostic factors. In multivariate analysis, age (P=0.0001), serum creatinine (P<0.002) and corticosteroid treatment (P=0.002) were independent prognostic variables. A more dramatic decrease of median serum bilirubin values (micromol/l) was observed at 7 and 14 days in corticosteroid patients (P<0.05) : -76.5 vs. -35 and -105 vs. -45. CONCLUSIONS: Corticosteroids improved short-term survival of patients with severe AH. Age and serum creatinine are independent prognostic factors. Corticosteroids are recommended for patients with severe AH. PMID- 11943419 TI - Reproducible production of thioacetamide-induced macronodular cirrhosis in the rat with no mortality. AB - BACKGROUND/AIMS: Hepatotoxin-induced rat models of liver cirrhosis are limited by the wide heterogeneity of cirrhosis produced. The present study developed a modified, reliable, and reproducible technique by which hepatic and systemic responses to thioacetamide during induction of cirrhosis were monitored by weekly weight changes. METHODS: Male Wistar rats (200-230 g) were divided into three groups. Group 1 (n=20) received continuous administration of 0.03% (w/v) thioacetamide in the drinking water for 12 weeks. Group 2 (n=20) received the same concentration of 0.03% thioacetamide as an initial concentration that was modified according to weekly weight changes in response to thioacetamide during the induction of cirrhosis. Group 3 (n=6) received normal water and served as controls. RESULTS: Mortality of Group 1 was 30% and the production of cirrhosis was only 45%. In contrast, there were no deaths in Group 2 and well-developed macronodular cirrhosis was found in 90% of the rats which was associated with significant portal hypertension, as indicated by increased portal venous pressure (13.6+/-0.4 vs. 9.1+/-0.3 mmHg, cirrhotic vs. control, respectively, P<0.01, Student's unpaired t-test). CONCLUSIONS: Variations in responses to thioacetamide can be easily monitored by weekly weight changes to reduce mortality to zero and simultaneously increase the production and quality of cirrhosis induced in rats. PMID- 11943420 TI - MELD score is better than Child-Pugh score in predicting 3-month survival of patients undergoing transjugular intrahepatic portosystemic shunt. AB - BACKGROUND/AIMS: Patients undergoing transjugular intrahepatic portosystemic shunt (TIPS) are at risk of early death due to end-stage liver failure. The aim of this study was to compare model of end-stage liver disease (MELD) and Child Pugh scores as predictors of survival after TIPS. METHODS: We studied 140 cirrhotic patients treated with elective TIPS. Concordance (c)-statistic was used to assess the ability of MELD or Child-Pugh scores to predict 3-month survival. The prediction of overall survivals was estimated by comparing actuarial curves of subgroups of patients stratified according to either Child-Pugh scores or MELD risk scores. RESULTS: During a median follow-up of 23.7 months, 55 patients died, 14 underwent liver transplantation and seven were lost to follow-up. For 3-month survival, the discrimination power of MELD score was superior to Child-Pugh score (0.84 vs. 0.70, z=2.07; P=0.038). Unlike Pugh score, MELD score identified two subgroups of Child C patients with different overall survivals (P=0.027). The comparison between observed and predicted survivals showed that MELD score overrates death risk. CONCLUSIONS: MELD score is superior to Child-Pugh score as predictor of short-term outcome after TIPS. Its accuracy, however, decreases for long-term predictions. PMID- 11943421 TI - Failure of Lactobacillus spp. to prevent bacterial translocation in a rat model of experimental cirrhosis. AB - BACKGROUND/AIMS: Prophylaxis of spontaneous bacterial peritonitis in cirrhotic patients with norfloxacin is associated with emergence of quinolone-resistant Enterobacteriaceae. We investigated whether an alternative strategy with Lactobacillus prevents bacterial translocation and ascitic fluid infection in cirrhotic rats. METHODS: CCl(4)-induced cirrhotic rats with ascites (n=34) were allocated to treatment with oral Lactobacillus strain GG at 1-2 x 10(9) cfu/day for 8-10 days (group LGG) or milk (group MILK). In addition, 20 cirrhotic rats were given a single dose of 15 mg norfloxacin orally and then allocated to Lactobacillus (group NOR-LGG) or milk (group NOR-MILK). Ten healthy rats served as control. After sacrifice the cecal flora were analyzed and the prevalence of bacterial translocation and ascitic fluid infection assessed. RESULTS: Cecal colonization with Lactobacillus was achieved in 90% of treated rats. The prevalence of bacterial translocation to mesenteric lymph nodes was 10% in control rats and 93, 84, 70 and 100% in groups MILK, LGG, NOR-MILK and NOR-LGG, respectively (P>0.1 for comparison of treatment groups), the prevalence of ascitic fluid infection was 60, 32, 40 and 40% (P>0.1). Bacterial translocation of Lactobacillus was observed in 24% of rats treated. CONCLUSION: Lactobacilli fail to prevent bacterial translocation and ascitic fluid infection in experimental cirrhosis in spite of successful intestinal colonization. PMID- 11943422 TI - Octreotide ameliorates the increase in collateral blood flow during postprandial hyperemia in portal hypertensive rats. AB - BACKGROUND/AIMS: The aims of this study were to examine, in a conscious rat model of portal hypertension, the effect of postprandial splanchnic hyperemia on collateral blood flow and to determine whether octreotide has an effect on postprandial collateral flow changes. METHODS: In rats with portal vein ligation, pulsed-Doppler flowmeters were implanted chronically around the splenorenal venous shunt (SRS), which is the main spontaneous collateral vessel in the portal hypertensive rat and around the superior mesenteric artery (SMA). Changes in flow after a standard liquid meal gavage and after the administration of octreotide were examined in the rat under unanesthetized and unrestricted conditions. RESULTS: SRS flow increased significantly after gavage with a standard liquid meal (10.6+/-2.9%) compared to orogastric intubation alone (-6.5+/-2.1%) (P<0.01). Similar flow changes were observed in the SMA after liquid meal gavage. The subcutaneous administration of octreotide at a dose of 400 g/kg reduces basal SRS flow (-19.5+/-2.3%) and significantly attenuated the change in SRS flow after liquid meal gavage (-8.1+/-2.9%) compared to animals that received placebo (3.6+/ 4.1% and 27.8+/-7.6%, respectively) (P<0.05). CONCLUSION: These results demonstrate that, in an experimental model of prehepatic portal hypertension, postprandial splanchnic hyperemia results in an increase in collateral flow that can be ameliorated with the use of octreotide. PMID- 11943423 TI - Dyssynchronous electrical and mechanical systole in patients with cirrhosis. AB - BACKGROUND/AIMS: Previous investigations have shown a prolonged QT interval in some patients with cirrhosis. The aim of this study was to investigate the relation between electrical and mechanical systole in patients with different degrees of severity of cirrhosis. METHODS: Forty-eight patients with cirrhosis and portal hypertension, studied during a haemodynamic investigation, were compared to 17 controls. RESULTS: A prolonged QTc (above 0.440 s(1/2)) was found in 37% of the cirrhotic patients vs. 5.9% in the controls (P=0.03), and there was a correlation to liver dysfunction (P<0.02). A direct relation between QT and time of mechanical systole (tS) was observed in controls (r=0.58, P<0.01), and cirrhotic patients (r=0.44, P<0.002). In patients with a prolonged QTc interval, the difference between electrical and mechanical systole time was substantially longer than in patients with a normal QTc interval (0.078 vs. 0.031 s, P<0.005). The QT values were related to markers of hyperdynamic circulation (r=-0.48 to 0.56, P<0.05-0.001). CONCLUSIONS; Prolonged repolarization, as evidenced by prolonged QTc, is related to both impaired liver function and systemic circulatory dysfunction. In addition these patients have alterations in the cardiac excitation-contraction relation with compromised association between electrical and mechanical function. PMID- 11943424 TI - B and C hepatitis viruses, HLA-DQ1 and -DR3 alleles and autoimmunity in patients with hepatocellular carcinoma. AB - BACKGROUND/AIMS: Hepatitis B and C involvement in hepatocellular carcinoma has been well established, but as yet not that of the human lymphocyte antigen (HLA) complex. To study viral, HLA and tumour interrelationships, 105 patients were evaluated for prevalence of viral markers and 161 patients, including 99 of the previous ones, for HLA allele frequency; the other 52 patients served as controls. METHODS: Immunoassays, molecular assays, microlymphocytotoxicity. RESULTS: Positivity for hepatitis B surface antigen and/or hepatitis C antibodies in 89% cirrhotic, 44% non-cirrhotic vs. 92% control patients (cirrhotic; all hepatitis C antibody positives were viraemic). Recurrent HLA alleles: HLA-Cw7 and -DQ1 in cirrhotic and control patients, HLA-Cw7, -B8 and -DR3 in non-cirrhotic patients compared with healthy controls (Pc=0.0000074, 0.000025, 0.0025, 0.00027 and 0.043, respectively). CONCLUSIONS: Viral data suggest a high chronic infection rate for cirrhotic patients. Recurrent HLA-Cw7 is compatible with natural killer cell activity inhibition to virus-infected and tumour cells by HLA C molecules. Recurrent HLA-DQ1 and -DR3 suggest the existence of an autoimmune condition with cell destruction in cirrhotic and without cell destruction in non cirrhotic patients as a consequence of autoreactive DQ-restricted T-helper (Th)1 and DR-restricted Th2 cells response, respectively. HLA-B8-DR3 linkage disequilibrium was possible. Thus, autoimmunity may have contributed to hepatocellular carcinoma development in these patients. PMID- 11943425 TI - NF-kappaB activation in non-parenchymal liver cells after partial hepatectomy in rats: possible involvement in expression of heparin-binding epidermal growth factor-like growth factor. AB - BACKGROUND/AIMS: Nuclear factor (NF) B activation plays a critical role in the initiation of liver growth after partial hepatectomy (PH). However, the issue of where specifically NF-B is activated is unclear. We previously reported that heparin-binding epidermal growth factor-like growth factor (HB-EGF) is a hepatotrophic factor. The aims of this study were to identify NF-B-activated cells and to clarify their involvement in HB-EGF expression after PH. METHODS: Using rats, a two-thirds PH was performed, after which NF-B-activated cells and HB-EGF-positive cells were identified by Southwestern histochemistry or immunohistochemistry. NF-B binding activity and HB-EGF gene expression were analyzed in rat hepatocytes and non-parenchymal cells (NPC) in primary culture. RESULTS: NF-B was activated in Kupffer cells and sinusoidal endothelial cells prior to activation in hepatocytes. HB-EGF immunoreactivity was detected after NF B activation in the sinusoidal cells from those localized in the periportal zones of hepatic lobules. HB-EGF gene expression by tumor necrosis factor was accompanied by an increase in NF-B binding activity in NPC but not in hepatocytes in primary culture, which was abolished by pyrrolidine dithiocarbamate, an inhibitor of NF-B activation. CONCLUSIONS; NF-B was activated in NPC prior to activation in hepatocytes. NF-B activation may be involved in HB-EGF expression in NPC after PH. PMID- 11943426 TI - Changes in the pool of bile acids in hepatocyte nuclei during rat liver regeneration. AB - BACKGROUND/AIMS: To investigate changes in nuclear bile acids (BAs) during rat liver regeneration. METHODS: Nuclei were isolated from control rat livers and after two-thirds partial hepatectomy (PH). BAs in bile, liver homogenate and nuclei were measured by gas chromatography-mass spectrometry. Nuclear translocation of radiolabeled BAs was determined using fresh isolated hepatocytes from control donors. RESULTS: Liver BA concentrations were transiently reduced after PH. Relative increases in: -MCA at 1 day, deoxycholic acid at 7 days and cholic acid (CA) at 3 and 14 days were found. Nuclear BAs accounted for <0.5% of liver BAs. Contamination with cytosolic BAs during nuclei isolation was <4%. Unconjugated- and conjugated-CA were able to reach the nucleus with similar efficiency. The pattern of nuclear BAs--CA (80%) and ursodeoxycholic acid (UDCA) (8.5%) being the most abundant--did not match that found in liver or bile. A transient decrease in CA/UDCA ratio, in absence of significant change in total BA content, was observed in nuclei after PH. "Flat" BA species were only detected in homogenate, but not in nuclei, at 1 day after PH. CONCLUSIONS: BA pool in nuclei of rat hepatocytes, whose composition is different to that of total liver BA pool, undergoes important changes during liver regeneration. PMID- 11943427 TI - Serum hepatitis B virus DNA levels and liver histology in inactive HBsAg carriers. AB - BACKGROUND/AIMS: A recent NIH research workshop on hepatitis B virus (HBV) revisited the definition of healthy HBsAg carriers. The new definition inactive surface antigen (HBsAg) carriers includes an estimated serum HBV DNA level below 105 copies/ml. However, this cut-off value needs to be confirmed. METHODS: Eighty five consecutive patients, HBsAg-positive/HBeAg-negative with persistently normal alanine aminotransferase (ALT) and undetectable serum HBV DNA with standard assay (Versant HBV DNA Assay (bDNA), Bayer) were prospectively followed for 3.2+/-2.6 (range 0.5-11) years; 58 underwent a liver biopsy. Serum HBV DNA was quantified with a sensitive polymerase chain reaction assay (Cobas Amplicor HBV Monitor, Roche) (sensitivity 200 copies/ml), and liver histology was assessed using the Ishak scoring system. RESULTS: The median serum HBV DNA level was 1300 copies/ml (<200-179 x 10(3) copies/ml), 16% of the subjects had no detectable serum HBV DNA and 98% had levels below 10(5) copies/ml. Histologic lesions were mild (total score <7) in all cases. Loss of HBsAg was observed in three patients, three patients experienced a transient increase in ALT (<2 x upper limit of normal), and serum HBV DNA levels remained stable (1-6 years) in 97% of the 38 patients retested. CONCLUSIONS: In our study of inactive HBsAg carriers, the median serum HBV DNA level was 1300 copies/ml, the serum HBV DNA level was below 10(5) copies/ml in 98% of the patients, and remained stable; histological lesions were mild in all cases. PMID- 11943428 TI - Alcoholic hepatitis--glucocorticosteroids or not? PMID- 11943429 TI - Clinical utility in quantifying serum HBV DNA levels using PCR assays. PMID- 11943430 TI - Liver stem cells and model systems for liver repopulation. PMID- 11943431 TI - Combined liver-kidney transplantation in a 15-year-old boy with alpha1 antitrypsin deficiency. AB - Alpha1-antitrypsin (1-AT) deficiency is the most common genetic cause of liver disease in infants and children. The major clinical manifestations include liver disease (primarily in children) and emphysema in adults. For patients who progress to cirrhosis and liver failure, liver transplantation provides a metabolic cure for the deficiency and presumably prevents the associated complications. Several case reports in the pediatric literature describe glomerulonephritis in the setting of severe 1-AT deficiency, but this association is less well documented in adults. End-stage chronic kidney disease is a rare finding in the literature and kidney transplantation is the treatment of choice. We report on a 15-year-old boy with 1-AT deficiency and consequent end-stage liver disease and membranoproliferative glomerulonephritis rapidly progressing to renal failure, who successfully underwent combined liver-kidney transplantation. PMID- 11943432 TI - Hepatocellular carcinoma skull metastasis with both extradural and subcutaneous extension. PMID- 11943433 TI - Apoptosis in alcoholic hepatitis. PMID- 11943434 TI - Lack of birth defects after paternal exposure to lamivudine. PMID- 11943435 TI - Combination therapy of chronic viral hepatitis B with lamivudine and interferon: better than what? PMID- 11943437 TI - First meta-analysis of octreotide for variceal bleeding. A lost opportunity. PMID- 11943438 TI - The dangers of inferring treatment effects from observational data: a case study in HIV infection. AB - Several recent articles have implicitly questioned the need for randomized controlled trials, based on a comparison of treatment effects from observational studies and related randomized controlled trials. We present here a counterexample of a comparison of two antiretroviral drugs used in the treatment of HIV infection, in which the observational analysis gave a potentially misleading result. Examples such as this emphasize the need to regard randomized controlled trials as the primary mechanism for assessing therapeutic efficacy. PMID- 11943439 TI - Randomized, placebo-controlled, parallel group versus crossover study designs for the study of dementia in Parkinson's disease. AB - In studies of dementia, crossover designs are controversial, reflecting concerns about temporal stability of disease, confounding of treatment effects with period by treatment interactions and/or carryover effects. Carryover effects are differences in the lingering effect of treatments (placebo) into subsequent periods. In the context of a trial to study the effect of donepezil on dementia in patients with Parkinson's disease, we examine two-sequence crossover studies with two or four periods, and a four-sequence design with two periods. We quantify bias in estimated treatment effects due to carryover effects and explore the use of biased estimators in hypothesis testing. For hypothesis testing, type I error rates are valid if (1) repeated administration of treatment alters the outcome only for effective treatments and (2) carryover effects due to placebo following treatment periods are nonzero only for effective treatments. For crossover and parallel group designs, sample sizes are adjusted for reduced statistical power due to carryover effects and temporal changes in variance. For the proposed clinical study, we estimate that a single-period parallel group design with baselines would require 104 patients and take about 23 months to complete. A two-sequence, four-period parallel group design with baselines would require about 80 patients and about 20 months to complete. We conservatively assume a carryover effect of 50% of the treatment effect for a two-sequence four period crossover design. The estimated treatment effect for this model may underestimate the true treatment effect by up to 13%. The sample size/study length requirements are 28 patients or 12.4 months, respectively, a substantial saving over either parallel group design. The cost of allowing for carryover in the sample size calculation is about 1.2 months of study time. PMID- 11943440 TI - On the variability of covariate adjustment. experience with Koch's method for evaluating the absolute difference in proportions in randomized clinical trials. AB - Recently Koch et al. suggested an interesting approach for covariate adjustment in randomized clinical trials. Here we report our experience with the approach in Assent II, a large randomized trial comparing 30-day mortality rates of two thrombolytic treatments. Data from the Gusto-I study and a recent successor to the Assent II study will also be used. Further, we balance the advantage of covariate adjustment (with Koch's method) to the extra efforts and resources necessary. In this context we highlight the fact that there is a potential risk of using a covariate-adjusted analysis as primary analysis and show that covariate adjustment has the least effect when it is needed most. Analytic results will be given to quantify this risk and a simulation study illustrates our findings. PMID- 11943441 TI - Quality assurance of asthma clinical trials. AB - Accuracy and repeatability of spirometry measurements are essential to obtain reliable efficacy data in randomized asthma clinical trials. We report our experience with a centralized spirometry quality assurance program that we implemented in our phase III asthma trials. Six asthma trials of 4 to 21 weeks in duration were conducted at 232 clinical centers in 31 countries. Approximately 23,100 prebronchodilator and 13,700 postbronchodilator spirometry tests were collected from 2523 adult and 336 pediatric asthmatic patients. The program used a standard spirometer (the Renaissance spirometry system) with maneuver quality messages and automated quality grading of the spirometry tests. Each clinical center transmitted spirometry data weekly to a central database, where uniform monitoring of data quality was performed and feedback was provided in weekly quality reports. Seventy-nine percent of all patients performed spirometry sessions with quality that either met or exceeded American Thoracic Society standards and improved over time. Good-quality spirometry was associated with (1) less severe asthma; (2) active treatment; (3) infrequent nocturnal awakenings; (4) age above 15 years; and (5) low body weight. Maneuver-induced bronchospasm was rare. Good-quality spirometry was observed in multicenter asthma clinical trials that employed a standard spirometer and continuous monitoring. Both within and between-patient variability decreased. Spirometry quality improved with time as study participants and technicians gained experience. PMID- 11943442 TI - The Diabetes Prevention Program: recruitment methods and results. AB - The Diabetes Prevention Program (DPP) is a multicenter randomized controlled trial designed to test whether diet and exercise or medication can prevent or delay the onset of type 2 diabetes in persons with impaired glucose tolerance, who are at increased risk of the disease. This paper describes DPP recruitment methods, strategies, performance, and costs. The DPP developed an organizational structure for comprehensive management and continuous monitoring of recruitment efforts. The DPP utilized a variety of recruitment strategies, alone or in combination, and a stepped informed consent procedure leading to randomization. Studywide and clinic-specific recruitment data were monitored, analyzed, and used to modify recruitment approaches. DPP recruitment was completed slightly ahead of schedule, meeting goals for the proportion of women enrolled and nearly meeting goals for the proportion of racial/ethnic minorities. Clinics varied widely in the recruitment strategies they used, and these strategies also varied by participant age, gender, and race/ethnicity. Staff time devoted to recruitment averaged 86.8 hours per week per clinic, with the majority of effort by staff specifically assigned to recruitment. The number of staff hours required to recruit a participant varied by recruitment strategy. Recruitment cost (excluding staff cost) was about 1075 US dollars per randomized participant. The DPP experience offers lessons for those planning similar efforts: (1) a method for ongoing assessment and revision of recruitment strategies is valuable; (2) a range of recruitment strategies may be useful; (3) the most effective methods for recruiting potential subjects may vary according to the gender, age, and race/ethnicity of those individuals; (4) recruitment strategies vary in the amount of staff time required to randomize a participant; and (5) a stepped screening may make it easier to identify and recruit volunteers who understand the requirements of the study. PMID- 11943445 TI - A simulation approach to determining sample size and power for group sequential trials. PMID- 11943443 TI - The ethics of research on informed consent. PMID- 11943446 TI - Traditional design does not choose values around 33%. PMID- 11943447 TI - Research on informed consent: investigator-developed versus focus group-developed consent documents, a VA cooperative study. AB - In the Department of Veterans Affairs Cooperative Study (VACSP) #470, A Randomized, Multicenter, Controlled Trial of Multi-Modal Therapy in Veterans with Gulf War Illnesses, a substudy was designed with the primary objective of comparing the utility of an informed consent document developed by a focus group of Gulf War veterans (focus group-developed) to an informed consent document developed by the standard process involving the study investigators (investigator developed). In December 1998 a focus group of five Gulf War veterans convened at the coordinating center and developed a consent document during three sessions. The focus group used the investigator-developed consent document as a "starting point" and then modified it by consensus agreement. They also reviewed and modified the substudy's assessment questionnaire. Utility will be evaluated in 1092 veterans participating in the parent study, VACSP #470, by directly comparing selected patient-centered outcomes between those receiving the focus group-developed consent document versus those receiving the investigator developed document. The primary outcomes to be evaluated over a 1-year follow-up period include measures of the informed consent process, such as patient recall, expectations about risks and benefits of participation, and understanding about the voluntariness of consent. Secondary outcomes will assess the impact of the substudy on the parent study with respect to recruitment and adherence. VACSP #470 was initiated in May 1999 in 20 sites that were randomly allocated to use either the focus group-developed or investigator-developed consent document. Sites are unaware of the type of consent document assigned. This article focuses on the rationale and design of the informed consent substudy and also discusses potential ethical issues. PMID- 11943448 TI - The evaluation of subcutaneous proleukin (interleukin-2) in a randomized international trial: rationale, design, and methods of ESPRIT. AB - The Evaluation of Subcutaneous Proleukin in a Randomized International Trial (ESPRIT) is a large ongoing randomized trial of subcutaneous interleukin-2 (IL-2) plus antiretroviral therapy versus antiretroviral therapy alone in patients with HIV (human immunodeficiency virus) disease and CD4 cell counts of at least 300 cells/mm(3). The primary objective is to determine whether the addition of IL-2 to combination antiretroviral therapy improves morbidity and mortality. The aim is to recruit 4000 participants and follow them for an average of 5 years. Eligible subjects will be recruited at 275 investigational sites in 23 countries around the world. Coupled with broad eligibility criteria this will ensure widely applicable results. A range of secondary objectives will also be addressed in this setting that will include the conduct of observational studies and nested substudies with a public health focus. This article describes the rationale supporting the trial in addition to reviewing the study design, coordination, and governance. PMID- 11943449 TI - Electrophysiology of Necturus taste cells. AB - Taste buds are sensory end organs that detect chemical substances occurring in foodstuffs and relay the relative information to the brain. The mechanisms by which the chemical stimuli are converted into biological signals represent a central issue in taste research. Our understanding of how taste buds accomplish this operation relies on the detailed knowledge of the biological properties of taste bud cells-the taste cells-and of the functional processes occurring in these cells during chemostimulation. The amphibian Necturus maculosus (mudpuppy) has proven to be a very useful model for studying basic cellular processes of vertebrate taste reception, some of which are still awaiting to be explored in mammals. The main advantages offered by Necturus are the large size of its taste cells and the relative accessibility of its taste buds, which can therefore be handled easily for experimental manipulations. In this review, I summarize the functional properties of Necturus taste cells studied with electrophysiological techniques (intracellular recordings and patch-clamp recordings). My focus is on ion channels in taste cells and on their role in signal transduction, as well as on the functional relationships among the cells inside Necturus taste buds. This information has revealed to be well suited to outline some of the general physiological processes occurring during taste reception in vertebrates, including mammals, and may represent a useful framework for understanding how taste buds work. PMID- 11943450 TI - Costorage and coexistence of neuropeptides in the mammalian CNS. AB - The term neuropeptides commonly refers to a relatively large number of biologically active molecules that have been localized to discrete cell populations of central and peripheral neurons. I review here the most important histological and functional findings on neuropeptide distribution in the central nervous system (CNS), in relation to their role in the exchange of information between the nerve cells. Under this perspective, peptide costorage (presence of two or more peptides within the same subcellular compartment) and coexistence (concurrent presence of peptides and other messenger molecules within single nerve cells) are discussed in detail. In particular, the subcellular site(s) of storage and sorting mechanisms within neurons are thoroughly examined in the view of the mode of release and action of neuropeptides as neuronal messengers. Moreover, the relationship of neuropeptides and other molecules implicated in neural transmission is discussed in functional terms, also referring to the interactions with novel unconventional transmitters and trophic factors. Finally, a brief account is given on the presence of neuropeptides in glial cells. PMID- 11943452 TI - Erratum. PMID- 11943453 TI - Mitochondria: evolution, genomics, homeostasis and pathology. Papers presented at an international meeting. May 12-20, 2001. Selva di Fasano (BR Italy). PMID- 11943451 TI - Pathogenic theories and intrathecal analysis of the sporadic form of Alzheimer's disease. AB - Alzheimer's disease (AD) is an age-dependent dementia characterized by progressive loss of cognitive functions and by characteristic pathological changes in the brain: the formation of aggregates extracellularly by beta-amyloid (Abeta) peptide and intracellularly by tau proteins. The disease presents several major diagnostic difficulties: (1) AD develops slowly; (2) analysis of damaged brain tissues is difficult, requiring a biopsy which poses ethical problems; (3) no biochemical markers are available for the diagnosis and monitoring of the disease progression. Since the cerebrospinal fluid (CSF) is in contact with the extracellular space of the brain, many studies have tried to correlate the levels of the intrathecal peptides and amino acids and the development of dementia. The present review analyzes the main results of intrathecal content analyses in light of pathogenic theories proposed to explain the damage associated with AD and observed in the brain of patients by postmortem examination. PMID- 11943454 TI - Mitochondrial DNA in metazoa: degree of freedom in a frozen event. AB - The mitochondrial genome (mtDNA), due to its peculiar features such as exclusive presence of orthologous genes, uniparental inheritance, lack of recombination, small size and constant gene content, certainly represents a major model system in studies on evolutionary genomics in metazoan. In 800 million years of evolution the gene content of metazoan mitochondrial genomes has remained practically frozen but several evolutionary processes have taken place. These processes, reviewed here, include rearrangements of gene order, changes in base composition and arising of compositional asymmetry between the two strands, variations in the genetic code and evolution of codon usage, lineage-specific nucleotide substitution rates and evolutionary patterns of mtDNA control regions. PMID- 11943455 TI - Amino acid replacement is rapid in primates for the mature polypeptides of COX subunits, but not for their targeting presequences. AB - We examined inferred amino acid replacements for 16 genes that encode the proteins of the cytochrome c oxidase (COX) holoenzyme in eight vertebrate species. Phylogeny-based analysis revealed that the human lineage (primates) has had an unusually large, statistically significant, number of amino acid replacements in the mature protein coding region of these genes. This finding is similar to earlier observations of an accelerated non-synonymous substitution rate for some lineages of primates for COX1, COX2, COX4, and COX7AH. In contrast, the mitochondrial targeting presequences of these same proteins have not undergone a concomitant rate change. This more comprehensive analysis suggests that COX5A, COX6B, COX6C, COX7C, and COX8L have also undergone an acceleration in amino acid replacement rates in anthropoid primates. Some of these rate accelerations (e.g. in COX5A and COX7C) are so pronounced that non-human mammalian sequences are more similar to sequences from Xenopus or zebrafish than they are to human. Since the functions of the targeting and mature proteins of these polypeptides are different, the mature portions of these genes are likely to have undergone a functionally significant change that is adaptive in nature. PMID- 11943457 TI - Transcription of two sunflower (Helianthus annuus L.) mitochondrial tRNA genes having different genetic origins. AB - The divergent transcription of two tRNA genes encoded in sunflower mitochondrial DNA, proposed as genes of different genetic origin, has been studied in detail. The transcription initiation site (TIS) for both transcript precursors has been identified by hybridization with in vitro (32)P-capped total RNAs and primer extension. The location of two TISs and the analysis of distribution of sequence elements (motifs) usually present in higher plant mitochondrial promoters led to the identification of two short regions (about 30-40 bp) which can be proposed as the promoters for the transcription of two genes. This conclusion is supported by the observation that within the short intergenic region included between the 5' termini of two genes (1924 bp) the distribution of those specific motifs is unique around the TISs, although not identical for the two promoters. Based on specific experimental results the trnE promoter shows a higher efficiency in comparison with that of the trnH promoter. This result is in good agreement with its structure which strictly conforms to those described for mitochondrial genes of dicot plants. Instead the other promoter shows some divergences which could be responsible for its lower efficiency. The context in which trnH lies in the sunflower mitochondrial genome and other features described in the paper may suggest that, despite the high similarity with the chloroplast counterpart, the trnH gene could have a native origin. PMID- 11943456 TI - Role of editing in plant mitochondrial transfer RNAs. AB - Editing in plant mitochondria consists in C to U changes and mainly affects messenger RNAs, thus providing the correct genetic information for the biosynthesis of mitochondrial (mt) proteins. But editing can also affect some of the plant mt tRNAs encoded by the mt genome. In dicots, a C to U editing event corrects a C:A mismatch into a U:A base pair in the acceptor stem of mt tRNA(Phe) (GAA). In larch mitochondria, three C to U editing events restore U:A base pairs in the acceptor stem, D stem and anticodon stem, respectively, of mt tRNA(His) (GUG). For both these mt RNA(Phe) and tRNA(His), editing of the precursors is a prerequisite for their processing into mature tRNAs. In potato mt tRNA(Cys) (GCA), editing converts a C28:U42 mismatch in the anticodon stem into a U28:U42 non-canonical base pair, and reverse transcriptase minisequencing has shown that the mature mt tRNA(Cys) is fully edited. In the bryophyte Marchantia polymorpha this U residue is encoded in the mt genome and evolutionary studies suggest that restoration of a U28 residue is necessary when it is not encoded in the gene. However, in vitro studies have shown that neither processing of the precursor, nor aminoacylation of tRNA(Cys), requires C to U editing at this position. But sequencing of the purified mt tRNA(Cys) has shown that Psi is present at position 28, indicating that C to U editing is a prerequisite for the subsequent isomerization of U into Psi at position 28. PMID- 11943458 TI - Striking differences in RNA editing requirements to express the rps4 gene in magnolia and sunflower mitochondria. AB - The ribosomal protein S4 gene (rps4) has been identified as a single copy sequence in the mitochondrial genomes of two distant higher plants, Magnolia and Helianthus. Sequence analysis revealed that the rps4 genes present in the magnolia and sunflower mitochondrial genomes encode S4 polypeptides of 352 and 331 amino acids, respectively, longer than their counterparts in liverwort and bacteria. Expression of the rps4 genes in the investigated higher plant mitochondria was confirmed by Western blot analysis. In Helianthus, one of two short nucleotide insertions at the 3'-end introduces in the coding region a premature termination codon. Northern hybridizations and reverse transcription polymerase chain reaction analysis demonstrated that the monocistronic RNA transcripts generated from the rps4 locus in Magnolia and Helianthus mitochondria are modified by RNA editing at 28 and 13 positions, respectively. Although evolutionarily conserved, RNA editing requirements of the rps4 appear more extensive in Magnolia than in Helianthus and in the other higher plants so far investigated. Furthermore, our analysis also suggests that selection of editing sites is RNA sequence-specific in a duplicated sequence context. PMID- 11943459 TI - Mitochondrial effects of the pleiotropic proteasomal mutation mpr1/rpn11: uncoupling from cell cycle defects in extragenic revertants. AB - We have previously characterized a Saccharomyces cerevisiae mutant which contains a mutation in the essential rpn11/mpr1 gene coding for the proteasomal regulatory subunit Rpn11. The mpr1-1 mutation shows the phenotypic characteristics generally associated with proteasomal mutations, such as cell cycle defects and accumulation of polyubiquitinated proteins. However, for the first time, mitochondrial defects have also been found to be a consequence of a mutation in a proteasomal gene (Mol. Biol. Cell 9 (1998) 2917-2931). Since the mutant strain is thermosensitive both on glucose and on glycerol, we searched for revertants in order to shed light on the Rpn11/Mpr1 functions. Spontaneous revertants able to grow on glucose but not on glycerol at 36 degrees C were isolated, and, only from them, revertants able to grow at 36 degrees C on glycerol were selected. Revertants of the two classes were found to be extragenic. The detailed characterization of these extragenic suppressors demonstrates that the phenotypes related to cell cycle defects can be dissociated from those concerned with mitochondrial organization. PMID- 11943460 TI - Cytochrome oxidase in health and disease. AB - Yeast and bovine cytochrome c oxidases (COX) are composed of 12 and 13 different polypeptides, respectively. In both cases, the three subunits constituting the catalytic core are encoded by mitochondrial DNA. The other subunits are all products of nuclear genes that are translated on cytoplasmic ribosomes and imported through different transport routes into mitochondria. Biogenesis of the functional complex depends on the expression of all the structural and more than two dozen COX-specific genes. The latter impinge on all aspects of the biogenesis process. Here we review the current state of information about the functions of the COX-specific gene products and of their relationship to human COX deficiencies. PMID- 11943461 TI - Mitochondrial functions and aging. AB - In the filamentous ascomycete Podospora anserina mitochondria play a major role in lifespan control. Since the function of these organelles depends on a large number of individual components it is no surprise that a complex network of interacting branches of individual molecular pathways is involved in this process. Recently, the nuclear encoded transcription factor GRISEA was found to significantly affect mitochondrial functions. GRISEA is involved in the control of cellular copper homeostasis. Most importantly, the high affinity uptake of copper from the environment is controlled by this transcription factor. Once copper has entered the cell, it becomes distributed to different compartments and different target molecules. This process depends on a group of proteins, termed copper chaperones. PaCOX17, a homologue of the yeast copper chaperone yCOX17, appears to be involved in copper delivery to mitochondria. Most importantly, the metal is crucial for the assembly and the function of complex IV of the respiratory chain. However, although P. anserina is an obligate aerobe and therefore depends on mitochondrial energy transduction, impairments in the copper delivery pathway are not lethal. This is due to the induction of a molecular back up system able to compensate for deficiencies in complex IV. The system utilizes an alternative oxidase (PaAOX) which uses iron instead of copper as a cofactor. The alternative respiratory pathway is characterized by a decreased ATP generation but, most significantly, also a decrease in the production of reactive oxygen species. Consequently, molecular damage is reduced which contributes to an increased lifespan of this type of mutant. In addition, modifications in the availability of cellular copper have other relevant consequences. Most significantly, the characteristic age-related rearrangements occurring in the mitochondrial DNA of wild-type strains of P. anserina were found to be dependent on the availability of copper. PMID- 11943462 TI - Evolution of a protein-rich mitochondrial ribosome: implications for human genetic disease. AB - Mitochondrial ribosomes comprise the most diverse group of ribosomes known. The mammalian mitochondrial ribosomes (55S) differ unexpectedly from bacterial (70S) and cytoplasmic ribosomes (80S), as well as other kinds of mitochondrial ribosomes. The bovine mitochondrial ribosome has been developed as a model system for the study of human mitochondrial ribosomes to address several questions related to the structure, function, biosynthesis and evolution of these interesting ribosomes. Bovine mitochondrial ribosomal proteins (MRPs) from each subunit have been identified and characterized with respect to individuality and electrophoretic properties, amino acid sequence, topographic disposition, RNA binding properties, evolutionary relationships and reaction with affinity probes of ribosomal functional domains. Several distinctive properties of these ribosomes are being elucidated, including their antibiotic susceptibility and composition. Mammalian mitochondrial ribosomes lack several of the major RNA stem structures of bacterial ribosomes but they contain a correspondingly higher protein content (as many as 80 proteins), suggesting a model where proteins have replaced RNA structural elements during the evolution of these ribosomes. Despite their lower RNA content they are physically larger than bacterial ribosomes, because of the 'extra' proteins they contain. The extra proteins in mitochondrial ribosomes are 'new' in the sense that they are not homologous to proteins in bacterial or cytoplasmic ribosomes. Some of the new proteins appear to be bifunctional. All of the mammalian MRPs are encoded in nuclear genes (a separate set from those encoding cytoplasmic ribosomal proteins) which are evolving more rapidly than those encoding cytoplasmic ribosomal proteins. The MRPs are imported into mitochondria where they assemble coordinately with mitochondrially transcribed rRNAs into ribosomes that are responsible for translating the 13 mRNAs for essential proteins of the oxidative phosphorylation system. Interest is growing in the structure, organization, chromosomal location and expression of genes for human MRPs. Proteins which are essential for mitoribosome function are candidates for involvement in human genetic disease. PMID- 11943463 TI - Transcriptional activators and coactivators in the nuclear control of mitochondrial function in mammalian cells. AB - The biogenesis and function of mitochondria rely upon the regulated expression of nuclear genes. Recent evidence points to both transcriptional activators and coactivators as important mediators of mitochondrial maintenance and proliferation. Several sequence-specific activators including NRF-1, NRF-2, Sp1, YY1, CREB and MEF-2/E-box factors, among others, have been implicated in respiratory chain expression. Notably, recognition sites for NRF-1, NRF-2 and Sp1 are common to most nuclear genes encoding respiratory subunits, mitochondrial transcription and replication factors, as well as certain heme biosynthetic enzymes and components of the protein import machinery. Moreover, genetic evidence supports a role for NRF-1 in the maintenance of mtDNA during embryonic development. Despite these advances, the means by which multiple transcription factors are integrated into a program of mitochondrial biogenesis remains an open question. New insight into this problem came with the discovery of the transcriptional coactivator, PGC-1. This cofactor is cold inducible in brown fat and interacts with multiple transcription factors to orchestrate a program of adaptive thermogenesis. As part of this program, PGC-1 can up-regulate nuclear genes that are required for mitochondrial biogenesis in part through a direct interaction with NRF-1. Ectopic expression of PGC-1 induces the expression of respiratory subunit mRNAs and leads to mitochondrial proliferation in both cultured cells and transgenic mice. More recently, PRC was characterized as a novel coactivator that shares certain structural similarities with PGC-1 including an activation domain, an RS domain and an RNA recognition motif. However, unlike PGC-1, PRC is not induced significantly during thermogenesis but rather is cell-cycle regulated in cultured cells under conditions where PGC-1 is not expressed. PRC has a transcriptional specificity that is very similar to PGC 1, especially in its interaction with NRF-1 and in the activation of NRF-1 target genes. These regulated coactivators may provide a means for integrating sequence specific activators in the biogenesis and function of mitochondria under diverse physiological conditions. PMID- 11943464 TI - Differences in nuclear gene expression between cells containing monomer and dimer mitochondrial genomes. AB - It is known that point mutations and rearrangements (deletions and duplications) of mammalian mitochondrial DNA (mtDNA) can result in mitochondrial dysfunction and human disease. Very little attention has been paid to mtDNA circular dimers (a complex form consisting of two genomes joined head-to-tail) despite their close association with human neoplasia. MtDNA dimers are frequently found in human leukemia, but the clinical relevance of their presence remains unknown. To begin to investigate the role of circular dimer mtDNA in the tumorigenic phenotype, we have created isogenic cell lines containing monomer and dimer mitochondrial genomes and compared the respective nuclear mRNA expression using Affymetrix gene array analysis. Surprisingly, a large number of nuclear gene changes were observed, with one of the largest category of genes being associated with remodeling of the cell surface and extracellular matrix. Since cell growth, migration, apoptosis, and many other cellular processes are influenced by signals initiating from the cell surface, the changes associated with the presence of mtDNA dimers could lead to significant alterations in tumorigenic potential and/or progression. PMID- 11943465 TI - Characterization of the mtTFA gene and identification of a processed pseudogene in rat. AB - Mitochondrial DNA replication and transcription are regulated from essential nucleus-encoded components that interact with the mitochondrial (mt) D-loop region. Among these there is the mitochondrial transcription factor A (mtTFA or Tfam). We have determined the sequence of the cDNA mtTFA in rat and have demonstrated that the gene has a mosaic organization with six introns whose sizes we have calculated. A differential splicing transcript lacking exon 5 has been detected in all assayed tissues and represents 9.85% of the full length transcript. Beside the gene which is homologous to the one found in man and mouse, rat nuclear genome contains at least 12 copies of this gene or genome fragments with high similarity to mtTFA. We have determined the sequence of one of these copies. This resulted to have 76.26% similarity to the active gene but to lack introns, suggesting it might be a processed pseudogene. RT-PCR experiments have demonstrated that this pseudogene (psi mtTFA) is transcribed in liver tissue. PMID- 11943466 TI - Cloning of two sea urchin DNA-binding proteins involved in mitochondrial DNA replication and transcription. AB - The cloning of the cDNA for two mitochondrial proteins involved in sea urchin mtDNA replication and transcription is reported here. The cDNA for the mitochondrial D-loop binding protein (mtDBP) from the sea urchin Strongylocentrotus purpuratus has been cloned by a polymerase chain reaction based approach. The protein displays a very high similarity with the Paracentrotus lividus homologue as it contains also the two leucine zipper-like domains which are thought to be involved in intramolecular interactions needed to expose the two DNA binding domains in the correct position for contacting DNA. The cDNA for the mitochondrial single-stranded DNA-binding protein (mtSSB) from P. lividus has been also cloned by a similar approach. The precursor protein is 146 amino acids long with a presequence of 16 residues. The deduced amino acid sequence shows the highest homology with the Xenopus laevis protein and the lowest with the Drosophila mtSSB. The computer modeling of the tertiary structure of P. lividus mtSSB shows a structure very similar to that experimentally determined for human mtSSB, with the conservation of the main residues involved in protein tetramerization and in DNA binding. PMID- 11943467 TI - An allele of HRAS1 3'variable number of tandem repeats is a frailty allele: implication for an evolutionarily-conserved pathway involved in longevity. AB - The human HRAS1 belongs to an evolutionarily-conserved family of genes which enrolls among its members the yeast RAS2, a gene which regulates stress response and longevity in the Saccharomyces cerevisiae. In this paper we report that the frequency of the a3 allele of HRAS1 3'variable number tandem repeat (HRAS1 3'VNTR) decreases in centenarians in respect to young people, and we estimate that during aging a3 carriers have a relative mortality risk of 1.126 (95% CI=1.044-1.213). We propose that the germ-line variability at the HRAS1 locus impacts on the individual's capacity to reach the extreme limits of human life span. Furthermore, we provide suggestive evidence that a3 HRAS1 3'VNTR allele and inherited variants of the mitochondrial genome (mtDNA haplogroups) do not affect independently human longevity, thus recalling the nucleus-mitochondrion interaction which regulates stress response and life-span in the yeast. PMID- 11943468 TI - Mitochondrial DNA repair of oxidative damage in mammalian cells. AB - Nuclear and mitochondrial DNA are constantly being exposed to damaging agents, from endogenous and exogenous sources. In particular, reactive oxygen species (ROS) are formed at high levels as by-products of the normal metabolism. Upon oxidative attack of DNA many DNA lesions are formed and oxidized bases are generated with high frequency. Mitochondrial DNA has been shown to accumulate high levels of 8-hydroxy-2'-deoxyguanosine, the product of hydroxylation of guanine at carbon 8, which is a mutagenic lesion. Most of these small base modifications are repaired by the base excision repair (BER) pathway. Despite the initial concept that mitochondria lack DNA repair, experimental evidences now show that mitochondria are very proficient in BER of oxidative DNA damage, and proteins necessary for this pathway have been isolated from mammalian mitochondria. Here, we examine the BER pathway with an emphasis on mtDNA repair. The molecular mechanisms involved in the formation and removal of oxidative damage from mitochondria are discussed. The pivotal role of the OGG1 glycosylase in removal of oxidized guanines from mtDNA will also be examined. Lastly, changes in mtDNA repair during the aging process and possible biological implications are discussed. PMID- 11943469 TI - Reactive oxygen species affect mitochondrial electron transport complex I activity through oxidative cardiolipin damage. AB - The aim of this study was to investigate the influence of reactive oxygen species (ROS) on the activity of complex I and on the cardiolipin content in bovine heart submitochondrial particles (SMP). ROS were generated through the use of xanthine/xanthine oxidase (X/XO) system. Treatment of SMP with X/XO resulted in a large production of superoxide anion, detected by acetylated cytochrome c method, which was blocked by superoxide dismutase (SOD). Exposure of SMP to ROS generation resulted in a marked loss of complex I activity and to parallel loss of mitochondrial cardiolipin content. Both these effects were completely abolished by SOD+catalase. Exogenous added cardiolipin was able to almost completely restore the ROS-induced loss of complex I activity. No restoration was obtained with other major phospholipid components of the mitochondrial membrane such as phosphatidylcholine and phosphatidylethanolamine, nor with peroxidized cardiolipin. These results demonstrate that ROS affect the mitochondrial complex I activity via oxidative damage of cardiolipin which is required for the functioning of this multisubunit enzyme complex. These results may prove useful in probing molecular mechanisms of ROS-induced peroxidative damage to mitochondria, which have been proposed to contribute to those pathophysiological conditions characterized by an increase in the basal production of reactive oxygen species such as aging, ischemia/reperfusion and chronic degenerative diseases. PMID- 11943470 TI - Depletion of mitochondrial DNA in the skeletal muscle of two cirrhotic patients with severe asthenia. AB - Qualitative and quantitative alterations of mitochondrial DNA (mtDNA) in the skeletal muscle from two patients with cirrhosis and severe asthenia have been studied. The 4977 bp (mtDNA(4977)) and the 7436 bp (mtDNA(7436)) mtDNA deletions, as well as other mtDNA deletions, revealed by long extension PCR (LX-PCR), were found in the two patients, whereas the 10,422 bp (mtDNA(10,422)) mtDNA deletion was absent. Altogether, the qualitative alterations of mtDNA in cirrhotic patients with severe asthenia were comparable to those of age-matched healthy individuals. The mtDNA content, on the contrary, was substantially decreased in both patients with respect to control. Such mtDNA depletion might be explained by an increased, disease-related, oxidative damage to mtDNA, which probably affects the replication of the mitochondrial genome as already suggested in other oxidative stress-associated diseases. PMID- 11943471 TI - Mutations in human nuclear genes encoding for subunits of mitochondrial respiratory complex I: the NDUFS4 gene. AB - Among the mitochondrial disorders, complex I deficiencies are encountered frequently. Although some complex I deficiencies have been associated with mitochondrial DNA mutations, in the majority of the complex I-deficient patients mutations of nuclear genes are expected. This review attempts to summarize genetic defects affecting nuclear encoded subunits of complex I reported to date focusing on those found in the NDUFS4 gene. NDUFS4 product is 18 kDa protein which appears to have a dual role in complex I, at least: cAMP-dependent phosphorylation activates the complex; non-sense mutation of NDUFS4 prevents normal assembly of a functional complex in the inner mitochondrial membrane. PMID- 11943472 TI - The role of Rho GTPases in disease development. AB - The functionality and efficacy of Rho GTPase signaling is pivotal for a plethora of biological processes. Due to the integral nature of these molecules, the dysregulation of their activities can result in diverse aberrant phenotypes. Dysregulation can, as will be described below, be based on an altered signaling strength on the level of a specific regulator or that of the respective GTPase itself. Alternatively, effector pathways emanating from a specific Rho GTPase may be under- or overactivated. In this review, we address the role of the Rho-type GTPases as a subfamily of the Ras-superfamily of small GTP-binding proteins in the development of various disease phenotypes. The steadily growing list of genetic alterations that specifically impinge on proper Rho GTPase function corresponds to pathological categories such as cancer progression, mental disabilities and a group of quite diverse and unrelated disorders. We will provide an overview of disease-rendering mutations in genes that have been positively correlated with Rho GTPase signaling and will discuss the cellular and molecular mechanisms that may be affected by them. PMID- 11943473 TI - Genetic analyses of adaptin function from yeast to mammals. AB - Adaptor protein (AP) complexes are heterotetrameric assemblies of subunits named adaptins. Four AP complexes, termed AP-1, AP-2, AP-3, and AP-4, have been described in various eukaryotic organisms. Biochemical and morphological evidence indicates that AP complexes play roles in the formation of vesicular transport intermediates and the selection of cargo molecules for inclusion into these intermediates. This understanding is being expanded by the application of genetic interference procedures. Here, we review recent progress in the genetic analysis of the function of AP complexes, focusing on studies that make use of targeted interference or naturally-occurring mutations in various model organisms. PMID- 11943474 TI - Recoding: translational bifurcations in gene expression. AB - During the expression of a certain genes standard decoding is over-ridden in a site or mRNA specific manner. This recoding occurs in response to special signals in mRNA and probably occurs in all organisms. This review deals with the function and distribution of recoding with a focus on the ribosomal frameshifting used for gene expression in bacteria. PMID- 11943475 TI - Genomic structure and evolutionary context of the human feline leukemia virus subgroup C receptor (hFLVCR) gene: evidence for block duplications and de novo gene formation within duplicons of the hFLVCR locus. AB - In this paper we sought to analyze the genomic structure and context of human feline leukemia virus subgroup C receptor (hFLVCR), a human glucarate transporter like gene at chromosome 1q31, and compare it to that of a paralog (FLVCR14q) at chromosome 14q24. Splicing, polyadenylation, and expression patterns, as estimated by in silico analysis, differed between the two FLVCR genes despite their similar genomic structures, suggesting active and independent evolution of transcriptional and messenger RNA processing patterns after gene duplication. Promoter activity was bi-directional for hFLVCR, but not for its 14q paralog. The upstream 1q transcribed sequences were determined to comprise a novel gene of unknown function, LQK1. Annotation of contigs centered at hFLVCR and FLVCRL14q also revealed highly conserved gene clusters on chromosomes 1 and 14, inferred to result from a duplication. The clusters contained members of the FLVCR, Angel (KIAA0759), JDP, p21SNFT, and TGF- families, as well as two uncharacterized families. The genome-wide locations of both previously recognized and four de novo in silico predicted genes belonging to these seven families were determined. Phylogenetic analyses of these families were consistent with the hypothesis that the 1q/14q duplication occurred early within, or immediately prior to the vertebrate divergence, after the protostome-deuterostome divergence but before the amniote-amphibian divergence. PMID- 11943476 TI - Cloning of the murine non-muscle myosin heavy chain IIA gene ortholog of human MYH9 responsible for May-Hegglin, Sebastian, Fechtner, and Epstein syndromes. AB - Mutations in the non-muscle myosin heavy chain IIA gene (MYH9) are responsible for May-Hegglin anomaly, Sebastian, Fechtner and Epstein syndromes. These 'MYH9 related' diseases are inherited as an autosomal dominant trait and are characterized by a variable expressivity of clinical features, including macrothrombocytopenia, deafness, nephrites, cataract, and Dohle-like leukocyte inclusions. To gain information of the function of the non-muscle myosin heavy chain IIA protein (NMMHC-IIA), we have identified the murine orthologue Myh9 gene. The gene is localized in a region of chromosome 15 and encodes a predicted protein of 1960 amino acids. This protein shows a high homology to the human NMMHC-IIA with 98% identity. The Myh9 exon-intron junctions were deduced from a murine genomic clone that revealed a perfect conservation of the exon structure between the human and mouse gene. Myh9 is expressed in liver, kidney, lung, and spleen. A low level of transcripts was detected also in heart and brain while no expression was revealed in skeletal muscle and testis. In vertebrates, NMMHC-IIA shows a striking degree of homology to NMMHC-IIB, which is expressed at higher level in mouse brain and testis than in other tissues, confirming the hypothesis that the two non-muscle myosins have different functional roles within cells. PMID- 11943477 TI - A novel gene encoding a putative transmembrane protein with two extracellular CUB domains and a low-density lipoprotein class A module: isolation of alternatively spliced isoforms in retina and brain. AB - We report herein the cDNA cloning of a novel retina and brain specific gene from mouse and human encoding a putative transmembrane protein with an N-terminal signal sequence and two conserved extracellular CUB domains followed by a single copy of the low-density lipoprotein class A (LDLa) module. The mouse and human genes, termed NETO1 (neuropilin and tolloid like-1), display sequence identities of 87% at the nucleotide and 95% at the protein level. The human NETO1 gene comprises 13 exons on chromosome 18q22-q23 and gives rise to three different mRNA isoforms. Two alternative leader exons 1a and 1b generate transcripts that translate into putative signal peptides with individual sequence composition but otherwise do not affect the primary structure of the mature NETO1 protein. Usage of the internal exon 5 is restricted to the retinal tissue and generates a truncated transcript that codes for a putative soluble protein, termed sNETO1, with only one copy of the CUB domain while lacking the LDLa module. NETO1 exhibits 57% identity to the deduced amino acid sequence of a non-annotated nucleotide sequence in the GenBank database, therefore designated NETO2. Both NETO1 and NETO2 share an identical and unique domain structure thus representing a novel subfamily of CUB- and LDLa-containing proteins. The cytoplasmic domains of NETO1 and NETO2 are not homologous to other known protein sequences but contain a conserved FXNPXY-like motif, which is essential for the internalization of clathrin coated pits during endocytosis or alternatively, may be implicated in intracellular signaling pathways. PMID- 11943478 TI - The cloning and functional analysis of canine matrix metalloproteinase-13 gene promoter. AB - A fragment of the 5' untranslated region corresponding to the canine matrix metalloproteinase-13 (MMP-13), collagenase-3 gene promoter has been isolated and characterized in rat cardiocytes to investigate the role of MMP-13 in cardiac disease. The promoter fragment (1.5 kb) demonstrated regions of sequence homology with the collagenase gene promoter sequences already determined for other species. Conserved regions were identified and shown to correlate with DNA binding motifs including AP-1 sites, a nuclear factor (NF) B-like binding domain, GATA and Nkx2.5 sites. A consensus TATA box was identified and shown to direct transcription initiation approximately 27 bp upstream of the translation start site. The canine MMP-13 promoter fragment was sufficient to drive basal expression of a luciferase reporter gene in both Madin Darby canine kidney cells (MDCK) and primary rat cardiocytes. The activity of the promoter fragment could be significantly increased by the treatment of transfected primary rat cardiocytes with interleukin-1 (IL-1) and basic fibroblastic growth factor (bFGF), with some induction also observed with tumour necrosis factor (TNF). The canine MMP-13 promoter activity has also been compared to the basal and induced activity of the canine MMP-9, gelatinase B promoter in these cell types. PMID- 11943479 TI - Homologues of amino acid permeases: cloning and tissue expression of XAT1 and XAT2. AB - The L-type (LAT) family of amino acid transporters is composed of exchangers for neutral, cationic, and anionic amino acids. They form functional heterodimers with membrane glycoproteins, rBAT or 4F2hc/CD98, to which they are linked by a disulphide bond. We report the molecular cloning and tissue expression of new mouse and human homologues of the LAT family, termed mXAT1, mXAT2 and hXAT2. The latter two proteins may correspond to ortholog genes in mouse and human. The hXAT2 gene is located on chromosome 8q21.3. The cloned X amino acid transporter (XAT) cDNAs are predicted to encode proteins of about 50 kDa. From a phylogenetic point of view, the three XAT proteins cluster together, but sequence comparison and secondary structure prediction show that they are also related to the members of the LAT family. Like these transporters, the XAT proteins show 12 transmembrane domains and a conserved cysteine residue, located in the second extracellular loop. This conserved cysteine is involved in the disulphide bond formed between the known members of the LAT family and 4F2hc or rBAT. The mXAT1 and hXAT2 mRNAs are expressed in the kidney but they are not detectable in a variety of other tissues. The corresponding proteins were efficiently translated following transfection of their cDNAs in Chinese hamster ovary (CHO) cells. However, cDNA transfection in CHO cells did not induce amino acid uptake, even when cotransfected with vectors expressing 4F2hc or rBAT. This could be related to the fact that mXAT1 and hXAT2 did not form detectable disulphide-linked heterodimers with 4F2hc or rBAT when they were co-expressed in CHO cells. Identification of other putative partner(s) of these LAT family-related transporters may be necessary to understand their role in renal physiology. PMID- 11943480 TI - MERP1: a mammalian ependymin-related protein gene differentially expressed in hematopoietic cells. AB - We have utilized differential display polymerase chain reaction to investigate the gene expression of hematopoietic progenitor cells from adult bone marrow and umbilical cord blood. A differentially expressed gene was identified in CD34+ hematopoietic progenitor cells, with low expression in CD34- cells. We have obtained the full coding sequence of this gene which we designated human mammalian ependymin-related protein 1 (MERP1). Expression of MERP1 was found in a variety of normal human tissues, and is 4- and 10-fold higher in adult bone marrow and umbilical cord blood CD34+ cells, respectively, compared to CD34- cells. Additionally, MERP1 expression in a hematopoietic stem cell enriched population was down-regulated with proliferation and differentiation. Conceptual translation of the MERP1 open reading frame reveals significant homology to two families of glycoprotein calcium-dependant cell adhesion molecules: ependymins and protocadherins. PMID- 11943481 TI - Genomic organization of the siglec gene locus on chromosome 19q13.4 and cloning of two new siglec pseudogenes. AB - The sialic acid binding immunoglobulin-like lectin (Siglec) family of genes is a recently described member of the immunoglobulin superfamily. Within this Siglec family there is a subgroup of genes which bear a high degree of homology with Siglec-3 (CD33), thus designated the Siglec-3-like subgroup of Siglecs. While their mRNA structure has been reported, the full genomic organization of these genes, is not known. Genes of this subgroup have been mapped to chromosome 19q13.4, primarily through in situ hybridization. Through analysis of several bacterial artificial chromosome (BAC) clones, we studied an approximate 700 kb region that encompasses the putative Siglec gene locus on chromosome 19q13.4. We established the first detailed map of the locus, which contains 8 Siglec and Siglec-like genes. Our map shows the relative position of all genes and the precise distances between them, along with the direction of transcription of each gene. To our knowledge, this is the first report that describes the full genomic organization of all members of the CD33-like subgroup of Siglecs, including the promoter sequences of all genes. Members of this subfamily exhibit two patterns of organization of the signal peptide, which is followed by one V-set domain (except for the long form of the siglecL1 gene). Exons containing the C2-set domains are all comparable in size and are separated by linker exons. The transmembrane domain is encoded for by a separate exon of almost the same size in all genes. The total number of exons differs according to the number of C2-set Ig domains, but intron phases are identical. The cytoplasmic domain is always encoded by two exons. We further identified two new Siglec pseudogenes in this locus, and analyzed their tissue expression pattern and their structural features. PMID- 11943482 TI - Complex regulatory element within the gammaE- and gammaF-crystallin enhancers mediates Pax6 regulation and is required for induction by retinoic acid. AB - The paired domain, DNA-binding domain of Pax6 and other Pax transcription factors, is composed of two subdomains (PAI and RED), each recognizing distinct half-sites of the bipartite binding site in adjacent major grooves of the DNA helix. The alternatively spliced Pax6(5a) isoform containing 14 extra amino acids within the PAI domain recognizes the 5aCON sequence consisting of four interdigitated 5' half-sites of the bipartite consensus sequence. A genome database search for similar tetrameric Pax6(A) recognition sequences led to the identification of a Pax6-binding site in the lens-specific enhancer of the mouse E- and F-crystallin genes. This binding site combines the properties of bipartite and tetrameric recognition sequences and, by mutational analysis, is shown to mediate Pax6-dependent regulation of the E- and F-crystallin promoter constructs both in primary chicken lens cells and in chicken embryo fibroblasts. The Pax6 binding site is adjacent to a previously identified retinoic acid response element and is itself required for retinoic acid induction of the F- and E crystallin genes, suggesting that Pax proteins and retinoic acid receptors cooperate in transcriptional regulation. In summary, our protein-DNA binding and transactivation studies suggest that -crystallin genes are under the control of a multifunctional enhancer element that mediates Pax6 regulation as well as retinoic acid-mediated induction. PMID- 11943483 TI - Characterization of virginiamycin S biosynthetic genes from Streptomyces virginiae. AB - Streptomyces virginiae produces -butyrolactone autoregulators (virginiae butanolide, VB), which control the biosynthesis of virginiamycin M1 and S. A 6.3 kb region downstream of the virginiamycin S (VS)-resistance operon in S. virginiae was sequenced, and four plausible open reading frames (ORFs) (visA, 1,260 bp; visB, 1,656 bp; visC, 888 bp; visD, 1209 bp) were identified. Homology analysis revealed significant similarities with enzymes involved in the biosynthesis of cyclopeptolide antibiotics: VisA (53% identity, 65% similarity) to -lysine 2-aminotransferase (NikC) of nikkomycin D biosynthesis, VisB (66% identity, 72% similarity) to 3-hydroxypicolinic acid:AMP ligase of pristinamycin I biosynthesis, VisC (48% identity, 59% similarity) to lysine cyclodeaminase of ascomycin biosynthesis, and VisD (43% identity, 56% similarity) to erythromycin C 22 hydroxylase of erythromycin biosynthesis. Northern blotting as well as high resolution S1 analysis of the ORFs revealed that they were transcribed as two bicistronic transcripts, namely 3.0-kb visB-visA and another 2.7-kb visC-visD transcript, with promoters locating upstream of visB and visC, respectively. Transcription of the two operons was observed only 1 h after the VB production, which was 2 h before the virginiamycin production. Furthermore, prompt induction of the transcription was observed as a result of external VB addition, suggesting that the expression of the two operons was under the control of VB. PMID- 11943484 TI - Complete genomic structure of human rpS3: identification of functional U15b snoRNA in the fifth intron. AB - Analysis of the complete genomic structure of the human ribosomal protein S3 (rpS3) gene revealed the presence of a functional U15b snoRNA gene in its intron. Human ribosomal protein S3 (rpS3) gene of 6115 bp long has been identified to contain six introns and seven exons in this study. The first and fifth introns of human S3 gene contain functional U15 snoRNA genes. Although Xenopus and Fugu counterparts also have six introns and seven exons, S3 gene of Fugu contains two functional U15 snoRNAs in the fourth and sixth introns and two pseudo genes for U15 snoRNAs in the first and fifth introns. In Xenopus S1 gene encoding ribosomal protein S3, however, three of its six introns contain U15 snoRNA gene sequence. Sequence comparison of the U15 genes from Xenopus, Fugu and human revealed that the regions involved in binding to 28S rRNA and the consensus sequence (C, D and D' boxes) for snoRNAs are highly conserved among those genes from these three species. Human U15a and U15b RNAs which are derived from the first and the fifth introns, respectively, have been identified to be functional by microinjection of human U15a and U15b snoRNAs into Xenopus oocyte. Northern blot and primer extension analyses confirm that human U15b snoRNA is expressed in vivo. PMID- 11943485 TI - cDNA cloning and genomic organization of the murine MRP7, a new ATP-binding cassette transporter. AB - Cellular resistance to cytotoxic drugs is a major obstacle to the treatment of disseminated cancers. Multidrug resistance protein (MRP) subfamily is a member of the ATP-binding cassette transporters which has been shown to cause multidrug resistance, except for P-glycoprotein. A new MRP subfamily gene, mrp7A (Abcc10), and its splicing variant, mrp7B, were isolated from mouse. The lengths of the open reading frames of mouse mrp7A and mrp7B are 4383 and 4506 bp, respectively. Estimated polypeptide sequences of mrp7A and mrp7B are 1460 and 1501 amino acids. The mouse mrp7 gene consists of at least 21 exons and 20 introns spanning around 20 kb that is almost the same as the one in human MRP7 gene, but different with the other MRP subfamily genes. The promoter region was isolated from the genomic clone and shown to support the luciferase activity seven fold over the promoterless negative control and two fold activity higher than the positive control of SV40 promoter. The analysis of tissue expression of mrp7A and mrp7B showed that these two transcripts express differentially in specific tissues. PMID- 11943486 TI - Expression of Bcl-2 and Bax proteins in relation to quality of bovine oocytes and embryos produced in vitro. AB - The mechanisms underlying the visual assessment and selection of immature oocytes resulting in optimum embryonic development following in vitro maturation, fertilization and culture (in vitro maturation (IVM)/in vitro fertilization (IVF)/in vitro embryo culture (IVC)) are unknown. Also, the reasons for the more frequent occurrence of cytoplasmic fragmentation in in vitro produced bovine embryos, resulting in poor survival following cryopreservation and decreased pregnancy rates following embryo transfer are not clear. The objectives of this study are: (1) to investigate whether differences in the quality of immature oocytes and embryo fragmentation are associated with apoptosis; and (2) to study the pattern of Bcl-2 and Bax expression in oocytes and embryos to help elucidate their potential roles in the regulation of apoptosis during development. Bovine oocytes were obtained from slaughterhouse ovaries and divided into four grades (grades I-IV) based on their morphology. Oocytes of different grades were cultured in serum-free medium for 48h. Embryos were produced only from grade I oocytes (highest quality) via IVM, IVF and IVC procedures. The morphological analysis of apoptosis in oocytes and embryos was carried out using propidium iodide staining and terminal deoxynucleotidyl transferase mediated dUTP nick end labeling. The expression of Bcl-2 and Bax in oocytes and embryos of different qualities and stages was determined using western blotting. The results showed that the number of morphologically abnormal oocytes with shrinkage and/or fragmentation of the ooplasm, which are typical features of apoptosis, was significantly higher in grade IV oocytes (denuded oocytes, the lowest quality) than in grade I oocytes after 48h in vitro culture (P<0.05). DNA fragmentation, a hallmark of the biochemical changes seen in apoptotic cell death, was observed in morphologically fragmented oocytes and embryos. The expression of Bcl-2 was high in good quality oocytes and embryos, low in fragmented embryos, and hardly detectable in denuded oocytes. In contrast, the expression of Bax was found in all types of oocytes and embryos with the highest expression in the denuded oocytes. This implies that the ratio of Bcl-2 to Bax may be used to gauge the tendency of oocytes and embryos towards either survival or apoptosis. Overall, our results show that apoptosis appears to be an underlying mechanism of bovine oocyte degeneration and embryo fragmentation. Interactions between the Bcl-2 family of proteins may play a critical role in pre-implantation embryo development. These findings could have important implications for improving IVF and related techniques. PMID- 11943487 TI - An in vitro study on spontaneous myometrial contractility in the cow during estrus and diestrus. AB - Myometrial smooth muscle strips were collected from slaughtered cows in estrus and diestrus. Longitudinal and circular smooth muscle strips were mounted in organ baths and after equilibration time and 2g preload, their physiologic contractility was recorded for 3h. Area under the curve (AUC), mean amplitude (MA) and frequency of contractions (F) were studied. Differences between cycle phases, between muscle layers and over the recorded time period were statistically evaluated. In the cow, physiologic contractility patterns (measured as AUC and MA) of circular versus longitudinal myometrial strips are always different during the 3h recording. Significant differences between estrus versus diestrus are only found for circular layers, but not for longitudinal layers. Significant differences over time are only found for longitudinal layers. PMID- 11943488 TI - Effects of environmental factors, age and genotype on sperm production and semen quality in Bos indicus and Bos taurus AI bulls in Brazil. AB - The effects of ambient temperature and humidity, month, age and genotype on sperm production and semen quality in AI bulls in Brazil were evaluated. Data from two consecutive years were analyzed separately. Seven Bos indicus and 11 Bos taurus bulls from one artificial insemination (AI) center were evaluated in Year 1 and 24 B. indicus and 16 B. taurus bulls from three AI centers were evaluated in Year 2. Ambient temperature and humidity did not significantly affect sperm production and semen quality, probably because there was little variation in these variables. Month accounted for less than 2% of the variation in sperm production and semen quality. Increased bull age was associated with decreased sperm motility (P<0.10) and increased minor sperm defects (P<0.001) in Year 1. B. indicus bulls had greater (P<0.005) sperm concentration than B. taurus bulls in both years (1.7 x 10(9)/ml versus 1.2 x 10(9)/ml in Year 1 and 1.6 x 10(9)/ml versus 1.2 x 10(9)/ml in Year 2, respectively). Ejaculate volume was not significantly affected by genotype in Year 1 (6.6 ml versus 6.9 ml in B. indicus and B. taurus bulls, respectively), but B. indicus bulls had greater (P<0.05) total (11.4x10(9) versus 8.2 x 10(9)) and viable (6.7 x 10(9) versus 4.9 x 10(9)) numbers of spermatozoa in the ejaculate than B. taurus bulls. In Year 2, B. taurus bulls had greater (P<0.05) ejaculate volume than B. indicus bulls (8.2ml versus 6.7 ml, respectively) and total and viable number of spermatozoa in the ejaculate were not significantly different between genotypes (10.3 x 10(9) versus 9.1 x 10(9) and 6.1 x 10(9) versus 5.4 x 10(9) in B. indicus and B. taurus bulls, respectively). Sperm motility was not significantly affected by genotype (mean, 59%). In Year 1, B. indicus bulls tended (P<0.10) to have more major sperm defects and had more (P<0.05) total sperm defects than B. taurus bulls (11.8% versus 8.7% and 13.6% versus 10.0%, respectively). In Year 2, B. indicus bulls tended (P<0.10) to have more total sperm defects than B. taurus bulls (16.2% versus 13.3%, respectively). In conclusion, neither ambient temperature and humidity nor month (season) significantly affected sperm production and semen quality. B. indicus bulls had significantly greater sperm concentration and B. taurus bulls had significantly fewer morphologically defective spermatozoa. PMID- 11943489 TI - In vitro culture and interferon-tau secretion by ovine blastocysts. AB - Embryos were collected surgically from superovulated ewes on days 7, 8, 9 and 10 (oestrus=day 0) to evaluate the long-term culture and interferon-tau (IFN-tau) secretion of ovine blastocysts. Embryos were cultured in 2 ml Dulbecco's modification of Eagle's medium (DMEM) supplemented with 15 mg/ml BSA in 5% CO(2) in air or DMEM without BSA in 5% CO(2), 7% O(2), and 88% N(2) at 39 degrees C, examined daily for morphological features and diameter and each day placed into fresh culture medium to enable daily measurement of IFN-tau secretion. Nine day-7 and two day-9 embryos were cultured in DMEM with BSA and nine continued to develop. The day-7 embryos reached a mean maximum diameter of 370.0+/-50.25 microm after 4 days in culture. Nineteen day-7, 12 day-8 and five day-10 embryos were cultured in DMEM without BSA but only six of the day-7 and one day-8 embryos survived for at least 7 days with the former reaching a mean maximum diameter on day 7 of 357+/-43.75 microm whereas all five day-10 embryos survived for at least 7 days reaching a mean maximum diameter on day 6 of 1038+/-155.8 microm. An anti viral assay and a ELISA for IFN-tau were developed. There was a considerable variation in the time of onset and amount of IFN-tau secreted that did not seem to be related to embryo morphology. Of 28 day-7 embryos cultured, 60.7% were secreting IFN-tau after 1 day of culture whereas 87.5% of day-8 embryos were secreting IFN-tau after 1 day in culture. The mean concentration of IFN-tau secreted by day-8 embryos after 1 day in culture (10.99+/-2.55 ng/ml) was not significantly different to day-7 embryos after 2 days in culture (8.8+/-1.75 ng/ml). PMID- 11943490 TI - Influence of classification levels of ram sexual activity on spring breeding ewes. AB - Ram lambs (7-8 months old) and mature rams (19-20 months old) were used to evaluate the effect of classification levels of male sexual performance on reproductive performance of ewes during spring breeding. In Exp. 1, sexually active ram lambs with high (1.8+/-0.3; n=5) and low (0.9+/-0.2; n=5) sexual performance scores (HP and LP; mean+/-S.E.M.) were used in single sire breeding pens. Ewes (n=305) were stratified by age and assigned to 10 pens for 34 days starting in late March. For Exps. 2 and 3, two replicates were conducted for 2 years with sexually active mature rams in a single sire mating scheme. For Exp. 2, HP rams (n=5) averaged 3.6+/-0.2 ejaculations and LP rams (n=7) 1.8+/-0.2 ejaculations for sexual performance scores based on nine, 30 min serving capacity tests (SCT). Polypay ewes (n=152 to 153 per year) were stratified by age and assigned to pens each year for 34-38 days starting in late March for years 1 and 2. For Exp. 3, HP rams (n=6) averaged 3.7+/-0.1 ejaculations and LP rams (n=10) 2.3+/-0.1 ejaculations for sexual performance scores based on 18, 30 min SCT. Polypay ewes (n=229 in year 3 and n=244 in year 4) were stratified by age and assigned to pens each year for 34 days starting in late March. In Exp. 1, lambing rates for ewes bred to HP versus LP ram lambs did not differ (65.8 versus 53.0; P=0.20). Prolificacy tended (P=0.06) to be increased by 0.1 lambs in ewes bred by LP ram lambs. Total number of lambs born per ewe present at lambing, and lambing distribution were not altered by HP and LP ram lambs. In Exp. 2, lambing rates for fall-lambing ewes bred to mature HP or LP rams did not differ (58.1 versus 60.1; P=0.78). In Exp. 3, lambing rates for fall-lambing ewes bred to mature HP or LP rams did not differ (74.3 versus 69.0; P=0.35). There was no difference (P>0.10) between years for Exp. 2 or Exp. 3, and mature HP and LP rams did not affect the other reproductive variables monitored. Analyses of the combined data for Exps. 2 and 3 indicated only a year difference (P<0.001) in lambing rates and total lambs born. Present studies indicate that different sexual performance classifications for ram lambs and mature rams did not alter lambing rates or distribution of lambing of Polypays bred in late March to April. These results indicate that HP and LP, sexually active, Polypay rams and ram lambs with average to high quality semen can provide a source of rams for spring breeding Polypays in ambient conditions and that there was no advantage to using HP over sexually active LP ram lambs or rams. PMID- 11943491 TI - FSH induced stimulation of catalase activity in goat granulosa cells in vitro. AB - Reactive oxygen species scavenging enzymes like catalase play diverse role in mammals. The presence of catalase in mammalian ovary is now well established. In the present investigation, changes in catalase activity in granulosa cells isolated from follicles at various stages of differentiation in response to FSH were studied. The follicles were dissected out from goat ovaries and classified as small (<3mm), medium (3-6mm) or large (>6mm). Granulosa cells were isolated from categorized follicles. Results showed that there was a three-fold increase in catalase activity in granulosa cells from large follicles as compared to small and medium follicles. The catalase activity was stimulated significantly when granulosa cells were treated with FSH in vitro. The minimum effective dose that could stimulate catalase activity and estradiol secretion in case of granulosa cells from small and medium sized follicles was 100 ng/ml; for larger follicles, this value was 200 ng/ml. Concomitant to the increase in catalase activity, the estradiol secretion was significantly enhanced when cultured goat granulosa cells were treated with FSH. It was concluded that enzyme catalase may have a functional role in goat ovarian follicular development under endocrine regulation. PMID- 11943492 TI - Active immunization against gonadotrophin-releasing hormone in Chinese male pigs: effects of dose on antibody titer, hormone levels and sexual development. AB - The objective of this study was to determine the optimal dose of a GnRH vaccine for immunocastration of Chinese male pigs, based on immune, endocrine and testicular responses. Forty-two crossbred (Chinese Yanan x Large White) male pigs were randomly assigned to one of the five treatments as follows: (I) 0 microg (control, n=8); (II) 10 microg (n=8); (III) 62.5 microg (n=8); (IV) 125 microg (n=8); (V) 250 microg (n=10), D-Lys6-GnRH tandem dimer (TDK) peptide equivalent of conjugate (TDK-OVA), using Specol as the adjuvant. Pigs were immunized at 13 and 21 weeks of age and were slaughtered at 31 weeks of age. Blood samples for antibody titer and hormone assays were collected at 13, 21, 24 and 31 weeks of age. At these time-points, testis size was also measured. At slaughter, testis weight was recorded and fat samples were collected for androstenone assay. Four animals, one out of each immunized group, responded poorly to the immunization (non-responders). At slaughter, serum testosterone and LH levels, fat androstenone levels and testis size/weight of these non-responders were similar to those in control animals. Antibody titers of non-responders were substantially lower (P<0.05) than in other immunized pigs. For the animals that responded well to the immunization (immunocastrated pigs), serum testosterone and LH levels, fat androstenone levels and testis size or weight were reduced (P<0.05) as compared to either controls or non-responders, at all doses tested. There was a significant effect of dose of TDK-OVA on antibody titers. The overall mean antibody titers in the 62.5 or 125 microg dose group (53.6 and 50.5% binding, respectively) were significantly higher than in the 10 or 250 microg group (39.2 and 40.24% binding, respectively). At slaughter, there was a significant dose effect on testis size or weight and on serum testosterone levels, but there was no dose effect on serum LH levels and fat androstenone levels. Testis size or weight in the 10 microg group was reduced to a lesser extent (P<0.05) than in the three higher dose groups. At slaughter, in comparison to controls, mean testis size of immunocastrated pigs in treatments II-V was reduced to 55, 21, 33 and 25%, respectively, whereas testis weight was reduced to 39, 12, 18 and 14%, respectively. Reduction of testis size and/or weight is important for visual assessment of castration at the slaughterline, therefore, it is concluded that a dose of 10 microg peptide is not suitable. We conclude that, within the dose range studied, the 62.5 microg dose is optimal for future GnRH immunization studies or future practical use in immunocastration of Chinese male pigs. PMID- 11943493 TI - Activation of in vitro matured pig oocytes using activators of inositol triphosphate or ryanodine receptors. AB - In our study, we observed the activation of in vitro matured pig oocytes and their subsequent parthenogenetic cleavage after stimulation of ryanodine receptors (RyR) using ryanodine (Ry), caffeine or cyclic adenosine diphosphate ribose (cADPri) or after stimulation of inositol triphosphate receptors (IP(3)R) using D-myo-inositol 1,4,5-triphosphate (IP(3)). Heparin, a potent blocker of IP(3)R, prevented the activation of porcine oocytes using IP(3), but blockers of RyR (ruthenium red or procaine) prevented activation after stimulation by RyR and stimulation by IP(3)R using IP(3). The drugs were injected into oocytes matured to the stage of metaphase II and activation was determined by assessment of pronuclear formation. The activity of H1 kinase was determined and our results demonstrated a significant drop in H1 activity in the activated oocytes. The cleavage of parthenogenetic embryos progresses to more advanced stages after stimulation by IP(3)R than after stimulation by RyR. Our results could indicate that, in pig oocytes, the calcium released from IP(3)-sensitive stores triggers the calcium release from ryanodine-sensitive intracellular stores, which is necessary for oocyte activation. The calmodulin inhibitors ophiobolin A and W7 reduce the activation of oocytes induced by stimulation of RyR or IP(3)R. PMID- 11943494 TI - Estrogen-alpha and progesterone receptor expression in cystic endometrial hyperplasia and pyometra in the bitch. AB - Estrogen-alpha receptor (ER) and progesterone receptor (PR) were examined immunohistochemically in uteri of normal bitches, in uteri of bitches with cystic endometrial hyperplasia-mucometra (CEH-M) and in uteri of bitches with endometritis-pyometra (E-P), under exogenous progesterone treatment. In the CEH-M group, the ER- and PR-scores of all uterine cell types were higher than the ER- and PR-scores of normal uteri, although these differences were not always statistically significant. The ER-scores of E-P group were significantly lower than the ER-scores of the normal uteri and CEH-M group. The PR-scores of the E-P group tended to be higher than the PR-scores of the normal uteri, except for the surface epithelium, although these differences were not statistically significant. Exogenous progesterone treated bitches with CEH-M or E-P showed reduced ER- and PR-scores in the different uterine cell types, compared with the corresponding nontreated CEH-M or E-P group. The differences in ER and PR expression between CEH-M and E-P suggest different factors in the pathogenesis of both entities. Although, these changes in ER and PR expression do not seem to be directly involved in the pathogenesis of CEH-M and E-P. It is suggested that for CEH-M and progestin induced CEH-M a hormone dependent pathway is responsible. For P, the trigger may be bacterial infection. PMID- 11943495 TI - Gestational length in the koala, Phascolarctos cinereus. AB - We report a possible case of extended gestation in the koala, Phascolarctos cinereus. Birth of a pouch young was first observed 127 days after the removal of the male from a multi-female colony at Taronga Zoo. No other males were present at that time or had access to the facility. Head measurements and other growth data collected at the time of detection and over the period of pouch life indicates the time from removal of the male and the date of birth to be between 50 and 77 days. DNA fingerprinting using microsatellite loci unambiguously assigned paternity of the pouch young to this male. These observations suggest either an extended period of gestation of at least 50 days, or activation of a dormant blastocyst from the previous breeding season, as the female entered the period of seasonal oestrus. PMID- 11943497 TI - The pig platelet-activating factor receptor gene is expressed at the mRNA level in different tissues and is mapped to chromosome 6. AB - After the pig platelet-activating factor receptor (PAFr) gene was cloned and sequenced, the chromosomal location of this gene was studied using a pig/rodent somatic cell hybrid panel containing 27 cell lines. The results indicated that the pig PAFr gene is located on SSC6q22-23. Platelet-activating factor (PAF, 1-O alkyl-2-acetyl-sn-glycero-3-phosphocholine) is thought to be very important in the animal reproductive processes. Its function is mediated through a membrane bound receptor. Pig PAFr mRNA distribution in different tissues was tested using reverse transcription and PCR (RT-PCR) reactions. All tissues examined expressed PAFr. Using a pig PAFr gene DNA competitor, PAFr expression was quantificated. The pig PAFr mRNA expression level was estimated to be from 1 x 10(2) to 1.2 x 10(4) copies of complementary DNA (cDNA) per 50 ng of total RNA. The highest level was found in lung, and the lowest in the skeletal muscle. These results demonstrated that PAFr was differentially expressed in pig tissues. PMID- 11943496 TI - Parthenogenesis of rabbit oocytes activated by different stimuli. AB - Oocyte activation is one of the essential elements determining the success of nuclear transfer and the subsequent development of cloned embryos both in vitro and in vivo. Experiments were conducted to optimize the protocol for oocyte activation in a regular nuclear transfer study. In vivo derived oocytes were collected at 14-15 h from New Zealand white rabbits after ovulation treatment and were activated +18 h post-ovulation treatment. Single activation agents including calcium ionophore (A23187, 5 microM, 5 min), ethanol (Eth, 7%, 7 min), and thimerosal (200 microM, 10 min) were tested. Cleavage rates were highest in the ethanol-treated group (37%) compared to other treatments (19-25%). Very low blastocyst rates (2-3%) resulted which were not significantly different among treatments (P>0.05). Combined single agent treatment (calcium stimulators) with protein kinase inhibitor, 6-DMAP were used to achieve a full oocyte activation. Both pronuclear and blastocyst formation rates were significantly higher (P<0.05) in the Eth+6-DMAP treatment group (38 and 27%) than in the other groups (16-21 and 7-9%, respectively, P<0.05). Low (0.2mM) and high (2.5mM) concentrations of 6 DMAP treatments with different treatment lengths (1.5 and 3.5h) in the combined groups were also compared. Blastocyst formation and cleavage rates were greater in the high concentration with less treatment time groups (36% versus 4-20%, P<0.05). In conclusion, single activation agents, either Ca2+ stimulators or protein kinase inhibitors, could not fully activate mature rabbit oocytes. The best activation procedure obtained in this study was the Eth+6-DMAP combined treatment, which may be incorporated into regular nuclear transfer or cloning protocols. PMID- 11943498 TI - Viability and acrosome integrity of rabbit spermatozoa processed in a gelatin supplemented extender. AB - The effect of gelatin addition to the semen extender on the viability and acrosome integrity of rabbit spermatozoa was studied. Pooled semen samples were processed in a boar semen extender with or without gelatin addition. Semen samples were stored at 5 degrees C for 72 h. Viability and acrosome integrity was evaluated by light microscope. Results showed that gelatin addition had a significant positive effect on the quality of the stored semen. PMID- 11943499 TI - Superovulation using recombinant human FSH and ultrasound-guided transabdominal follicular aspiration in baboon (Papio anubis). AB - The response of baboon females to a modified human ovarian stimulation protocol incorporating start of pituitary suppression in the luteal phase of the cycle with a GnRH agonist (GnRHa) and recombinant human FSH (rhFSH) was studied. A long acting GnRHa implant supplying goserelin acetate was administered s.c. to six adult female baboons experiencing regular menstrual cycles (33-34 days) on days 22-24 of the cycle. Follicular development was monitored by transabdominal ultrasonography and serum levels of E2 and progesterone (P4) and rhFSH were determined by ELISA. Menses occurred 9-10 days after GnRHa administration. Daily i.m. administration of 75 IU rhFSH commenced 9-10 days after menses and continued for 9-10 days. When most follicles were > or =5mm diameter and serum E2 had reached its maximum level, 2000 IU hCG was administered i.m. to induce follicle maturation. Transabdominal ultrasound-guided follicular aspiration of follicles > or =2mm diameter was performed 30-34h after hCG administration. One baboon did not show an adequate response to rhFSH stimulation. This animal did not receive further treatment and no data for it are presented. The number of follicles aspirated was 21+/-4 and 17.2+/-3.8 oocytes were recovered per animal with an average recovery rate of 82% (86/105). The number of oocytes collected from five animals were 14, 21, 16, 15, and 20 (n=86). Most of the oocytes recovered were in metaphase II and 3h after recovery 91% (78/86) were considered suitable for in vitro fertilization. It was concluded that recombinant human FSH can successfully induce follicular recruitment and oocyte maturation in baboon females during pituitary suppression with a GnRHa PMID- 11943500 TI - Effects of cocaine/polydrug exposure and maternal psychological distress on infant birth outcomes. AB - To assess teratogenic effects of cocaine exposure and maternal psychological distress on birth outcomes, we conducted a longitudinal prospective study of 415 infants (218 cocaine-exposed--CE, 197 nonexposed--NE). Drug exposure was determined through a combination of maternal self-report, urine, and meconium screens. Maternal psychological distress postpartum was evaluated through a standardized, normative, self-report assessment. An extensive set of confounding variables was controlled, including severity of exposure to alcohol, tobacco, marijuana and other drugs, maternal age, race, parity, number of prenatal care visits, educational, marital, and socioeconomic status, and verbal and nonverbal intelligence. CE infants were smaller on all birth parameters and more likely to be preterm, small for gestational age, and microcephalic than NE infants. Forty one percent of cocaine users had clinically significant psychological symptoms, compared to 20% of a high-risk comparison group of noncocaine users. Consistent with a teratologic model, cocaine exposure independently predicted offspring birthweight, length, and head circumference. Maternal psychological distress self reported postnatally also independently predicted head circumference. Tobacco, alcohol, and marijuana exposures were also significant independent predictors of some fetal growth parameters. In addition, maternal distress symptoms, which may be reflective of maternal mental health disorders or responses to stress, added significantly to the risk for poorer fetal growth. PMID- 11943501 TI - Effects of perinatal buprenorphine and methadone exposures on striatal cholinergic ontogeny. AB - The effects of exposure to various doses of buprenorphine, methadone or water during the perinatal period were studied on striatal cholinergic development in the rat. Rats were exposed to buprenorphine (0.3 or 3.0 mg/kg/day), methadone (9 mg/kg/day) and/or water prenatally, postnatally or both pre- and postnatally via maternally implanted osmotic minipumps. The effects of buprenorphine varied with the dose used. There were some similarities between the effects of perinatal buprenorphine and perinatal methadone, such as a reduction in striatal acetylcholine (ACh) content in 4-day-old pups exposed prenatally to methadone or buprenorphine (0.3 mg/kg/day). However, differences were also observed between the effects of the two drugs. Unlike methadone, the 0.3-mg/kg/day dose of buprenorphine produced a sex-related increase in striatal ACh in male postnatal day (PND) 21 pups. The 3-mg/kg/day dose of buprenorphine produced a completely different range of results, such as decreased striatal ACh content in 4-day-old pups exposed to the drug postnatally and in 21-day-old pups exposed both pre- and postnatally. Differences in the effects of the two drugs may be related to the different affinities and efficacies of the drugs at different opioid receptor subtypes. PMID- 11943502 TI - Prenatal cocaine exposure induces an attenuation of uterine blood flow in the rat. AB - We have previously demonstrated that maternal cocaine injections result in a gradient of fetal brain cocaine levels that decrease as a function of the fetuses' proximity to the ovaries at embryonic (E) day 15. Our prior data suggest that cocaine-induced vasoconstriction may (1) limit cocaine's entry into the brain and (2) cause damage to DA neurons through injury associated with hypoxia or ischemia of the utero-placental junction. Therefore, using the microsphere technique (labeled with Ru(103)), the following study sought to determine whether the previously observed pattern of cocaine distribution among fetuses in the uterus were due to position-specific reductions in uterine or placental blood flow. On day 15, a single subcutaneous injection of 30 mg/kg cocaine HCl was administered to each rat. Thirty minutes after the cocaine injection, reference blood samples were drawn from the ventral tail artery. Uterine segments and placentae were removed and subjected to gamma counting. While results regarding placental blood flow were equivocal, cocaine significantly reduced average uterine blood flow by 54.6%. In addition, as one moves more proximal to the ovaries, cocaine progressively attenuates blood flow in uterine tissue segments. These data support the hypothesis that the pattern of drug distribution and subsequent brain alterations from prenatal cocaine exposure in our previous reports are likely due to differences in uterine blood flow. PMID- 11943503 TI - Effect of developmental lead exposure on the expression of specific NMDA receptor subunit mRNAs in the hippocampus of neonatal rats by digoxigenin-labeled in situ hybridization histochemistry. AB - The N-methyl-D-aspartate (NMDA) receptor has an important role in learning and memory. In this study, the effects of chronic Pb exposure on NMDA receptor subunit mRNAs expression in the developing rat hippocampus were examined by digoxigenin (DIG)-labeled in situ hybridization. In Pb-exposed rats, hippocampal NR1-2a mRNA levels were significantly increased in CA1, CA4 and DG subfields at 15 days of age and CA4 and DG subfields at 20 days of age. The lead (Pb) exposure resulted in a significant increase of NR2D mRNA levels in CA1, CA2, CA4 and DG hippocampal subfields at 20 days of age. Hippocampal NR2A mRNA levels were significantly decreased in CA1, CA2, CA3 and DG subfields from 15-day-old Pb exposed rats and CA1, CA3 and DG subfields from 20-day-old Pb-exposed rats. Hippocampal NR3A mRNA levels were also significantly decreased in CA1, CA4 and DG subfields at 15 days of age and CA1 and DG subfields at 20 days of age in Pb exposed rats. The Pb-induced subunit specific changes in hippocampal NMDA receptor subunit mRNA expression may lead to abnormality of natural NMDA stoichiometry and interfere with the physiological function of the NMDA receptor in the development of defined neuronal connections. PMID- 11943504 TI - Prenatal cocaine effects on fear conditioning: exaggeration of sex-dependent context extinction. AB - Prenatal cocaine exposure results in deficits in sensory preconditioning, discrimination reversal, and spatial navigation, tasks that require input from the hippocampus. However, there are no previous studies concerning prenatal cocaine effects on contextual fear conditioning, another hippocampal-dependent task. The present experiments tested whether chronic subcutaneous administration of 40 mg/kg of cocaine HCl to pregnant rats, from gestational day (GD) 8 through 20 would lead to disruption of contextual fear conditioning in adult male and female offspring. Offspring of saline-injected/pair-fed and untreated dams served as controls. Experiment 1 used a one-trial context conditioning preparation. Rats received a 2-s, 1-mA footshock in either the test context or a novel context, or received no shock on the day prior to the no-shock test. Defecation and freezing were measures of fear. Experiment 2 used a multiple measures protocol to optimize detection of prenatal treatment effects and was preceded by an open-field test. Rats received a 2-s, 0.8-mA footshock or no shock once daily over 4 days of conditioning. During 3 days of extinction, access to an adjacent chamber enabled the observation of four additional measures of fear: side crossing, latency, nose crossing, and side-differential. There were gender-dependent effects of conditioning on freezing and the four added measures of fear. Males showed higher levels of context conditioning and extinguished more slowly than females. The measures of nose crossing and side-differential revealed that prenatal cocaine exposure exaggerated gender-specific effects of context conditioning. The effects of prenatal cocaine exposure on context extinction are sexually dimorphic. PMID- 11943505 TI - Maternal alcohol and adrenalectomy: asynchrony of stress response and forced swim behavior. AB - Fetal alcohol-exposed (FAE) rats exhibit heightened hormonal and behavioral stress responses, strikingly similar to those caused by exposure to elevated maternal corticosterone (CORT). Since alcohol increases maternal CORT, this study examined the effect of maternal adrenalectomy (ADX) on the CORT stress response and forced swim test (FST) behavior of the adult FAE offspring. Maternal ADX alone dramatically enhanced the CORT stress response of the offspring of pair-fed (PF) mothers but had no effect on the exaggerated CORT response to restraint stress observed in the FAE female. In contrast, maternal ADX reversed the increased immobility of FAE offspring in the FST of depressive behavior. These findings provide original evidence that stress hyper-reactivity and depressive behavior in the FAE offspring are mediated by separate developmental mechanisms. PMID- 11943506 TI - Long-lasting effects of prenatal MAM treatment on water maze performance in rats: associations with altered brain development and neurotrophin levels. AB - We previously reported that prenatal methylazoxymethanol (MAM) administered on days 11 and 12 of rat pregnancy induces structural changes in the cytoarchitecture of the hippocampal-entorhinal axis. We also showed that young and middle-aged prenatally treated MAM animals displayed changes in brain neurotrophin levels [Neurosci. Lett. 309 (2001) 113; Physiol. Behav. 71 (2000) 57.]. To continue this line of investigation, the working hypothesis adopted was that prenatal MAM administration, by interfering with limbic neurogenesis, could impair learning and memory ability of aged animals in the water maze. It was found that injection of MAM during early rat brain development induced deficits in both the acquisition and retention phases of the Morris maze. These behavioral changes were associated with significant changes in brain nerve growth factor (NGF) and brain-derived neurotrophic factor (BDNF), reduced choline acetyltransferase (ChAT) immunoreactivity in forebrain cholinergic neurons and loss of neuropeptide Y (NPY) immunodistribution in cells of the entorhinal cortex. This finding, as well as confirming previous studies showing that injection of prenatal MAM administration induces significant changes in hippocampal-entorhinal axis neurogenesis and marked behavioral deficits in adult life, provides additional experimental evidence supporting the hypothesis that loss of NGF and/or BDNF-receptive or producing cells can co-occur at the onset of neurodevelopmental disorders. PMID- 11943507 TI - Assessing the potential toxicity of MK-801 and remacemide: chronic exposure in juvenile rhesus monkeys. AB - The present experiment examined the effects of chronic exposure to either 0.1 or 1.0 mg/kg MK-801 [a selective N-methyl-D-aspartate (NMDA) receptor antagonist] or 20.0 or 50.0 mg/kg remacemide (an NMDA receptor antagonist which also blocks fast sodium channels) in juvenile rhesus monkeys. Endpoints were monitored to provide a general index of subjects' health and included measures of clinical chemistry, hematology, ophthalmology, spontaneous home-cage behavior, and peak drug plasma levels. In general, both drugs were well tolerated and produced no treatment related effects during 2 years of dosing and assessment. Periodic plasma drug level determinations provided limited evidence that both compounds may induce their own metabolism. The present results contrast sharply with previously reported effects of long-lasting impairments in the acquisition of incremental learning and in the development of color and position discrimination in these same subjects. These observations highlight the importance of collecting a broad range of toxicology data, including tests of cognitive function, to make comprehensive assessments of new drug safety. In the present case, the less obvious effects of these drugs on cognition defined the toxicologic response. PMID- 11943508 TI - Impaired cued delayed alternation behavior in adult rat offspring following exposure to 2,3,7,8-tetrachlorodibenzo-p-dioxin on gestation day 15. AB - This investigation used random ratio (RR) and cued delayed alternation procedures to examine the operant behavior of adult male and female rats following prenatal 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD). Offspring were exposed to a single maternal dose of 0.0, 0.06, 0.18 or 0.54 microg/kg po of TCDD on gestation day (GD) 15. For RR, adult subjects were trained to respond on one lever in a two lever chamber for food reinforcement. The response requirement was increased across sessions. Male offspring responded at higher rates than females regardless of RR value and prenatal exposure history. For delayed alternation, animals were required to alternate responses on both apparatus levers and to inhibit responding during randomly interpolated delay intervals. The performance of male and female offspring exposed to 0.18-microg/kg TCDD was significantly less accurate and this group committed more errors by responding during the delay intervals than the other exposure groups. A similar trend was observed in the 0.54- microg/kg group. Overall, response accuracy during the delayed alternation procedure was inversely related to delay length and tended to improve with experience. Interpretations of these outcomes include the possibility that TCDD interfered with the development of attentional processes, impaired response inhibition or promoted response perseveration despite the presence of cues, indicating changes in reinforcement contingencies. PMID- 11943509 TI - Effects of prenatal exposure to manganese on postnatal development and behavior in mice: influence of maternal restraint. AB - Manganese (Mn) is an essential trace element whose deficiency and excess have been reported to cause central nervous system (CNS) disturbances. On the other hand, during pregnancy, maternal stress has been shown to enhance the developmental toxicity of a number of metals. In this study, the maternal toxicity and developmental effects of a concurrent exposure to Mn and restraint stress were evaluated in mice. Pregnant animals were divided into three groups and received subcutaneous injections of manganese chloride tetrahydrate (MnCl2.4H2O) at 0, 1 and 2 mg/kg/day on Gestation Days 6-18. Each group was divided into two subgroups. Mice in one subgroup were subjected to restraint for 2 h/day on Days 6-18 of gestation. Pregnant mice were allowed to deliver, and pups were evaluated for physical and neuromotor maturation. Subsequently, adult mice were also evaluated for activity and learning. A significant increase in perinatal mortality was observed at 2 mg/kg/day Mn. A delay in some developmental landmarks (eye opening, testes descent) due to Mn exposure (2 mg/kg/day) was also seen in both restrained and unrestrained animals. No differences in motor resistance and coordination, or in learning at the passive avoidance test, were noted in adult mice. At the current Mn doses, combined exposure to Mn and stress during the prenatal period did not produce long-lasting effects on adult mice. PMID- 11943510 TI - Differential vulnerability of snake species to MPTP: a behavioral and biochemical comparison in ratsnakes (Elaphe) and watersnakes (Nerodia). AB - The synthetic neurotoxicant 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP) induces a Parkinsonian-like syndrome in humans and nonhuman primates, and also causes movement disorders in rodents, fish, amphibians and lizards. To date, the effects of MPTP have not been characterized in snakes. In this study, the behavioral and biochemical effects of MPTP were assessed in the black ratsnake Elaphe o. obsoleta and the banded watersnake Nerodia f. fasciata--species that display contrasting behavioral sensitivities to dopaminergic antagonists and to amphibian toxins. We report that MPTP induces depletion of norepinephrine and serotonin in fore, mid and hindbrain regions and depletion of dopamine in fore and midbrain regions in E.o. obsoleta. MPTP also induced a marked reduction in righting ability in E.o. obsoleta. In N.f. fasciata, norepinephrine and dopamine were depleted by MPTP in all three brain regions and serotonin was only significantly reduced in the forebrain. In contrast to E.o. obsoleta, N.f. fasciata demonstrated no behavioral disorders. This study demonstrates a behavioral and biochemical sensitivity to MPTP in E.o. obsoleta that differs from that in N.f. fasciata. The differential sensitivities to monoaminergic modulation may be related to the contrasting diets of these species. PMID- 11943512 TI - Postnatal growth, neurobehavioral and neurophysiologic changes of prenatal low dose beta-radiation from tritiated water in mice. AB - Pregnant adult C57BL/6J mice were randomly assigned to four groups: one sham and three irradiated groups that were exposed to -irradiation from tritiated water (HTO) by a single intraperitoneal injection on Day 12.5 of gestation. The offspring received cumulative doses of 0.036, 0.071, and 0.213 Gy, respectively. The litters were observed for postnatal growth (body weight, brain weight), the development of four physiologic milestones (pinna detachment, eye opening, testes decent, vaginal opening), the acquisition age of several reflexes (cliff avoidance, air righting) and sensory functions (auditory startle, thermal reflex), movement and coordination functions and activity (pivoting, foot splay, continuous corridor activity), learning and memory performance (shock avoidance, conditioning reflex), and the density of CA1-CA4 hippocampal pyramidal neurons. Modest but significant dose-dependent neuronal death and functional impairment were seen in both 0.071 and 0.213 Gy groups. In conclusion, even prenatal low dose beta-radiation may impair murine central nervous system (CNS) development suggesting the potential importance of minimizing environmental exposure during human pregnancy. PMID- 11943511 TI - Effects of perinatal picrotoxin and sexual experience on heterosexual and homosexual behavior in male rats. AB - The effects of perinatal picrotoxin (0.75 mg/kg) on heterosexual and homosexual behavior of male rats, sexually experienced or not, were studied. The following data were obtained: (1) at birth, body weight and anogenital distance were not modified by the treatment; (2) during lactation, both treatment and sex interfered with body weight as well as in adult age; (3) as experimental animals were trained, the heterosexual behavior was improved; (4) picrotoxin treatment reduced lordotic response of homosexual behavior in inexperienced male rats and (5) the heterosexual experience with female rats inhibited homosexual behavior of both experimental and control animals. These results suggest that perinatal maternal picrotoxin exposure improved heterosexual behavior in male rats and the sexual experience reveals this effect. In addition, picrotoxin did not induce feminization in experimental inexperienced rats. Finally, the sexual experience per se promotes changes in brain regions related to male behavioral and sexual aspects. PMID- 11943513 TI - Comparative effects of maternal prenatal and postnatal exposures to astemizole on reproductive parameters of rats. AB - The prenatal and postnatal effects of administration of a nonsedative antihistamine H1, astemizole (ATZ), were compared. ATZ (10 mg/kg) was injected daily into female Wistar rats throughout pregnancy (prenatal treatment) or during the first 6 days of lactation (postnatal treatment). Neither treatment modified dam body weight. Prenatal exposure reduced offspring body weight and lead to a latter expression of the vaginal opening of female offspring. In addition, a long term disruption of male reproductive behavior was observed, while female sexual behavior was not modified. The sexual activity index and the intromission frequency were increased in prenatally treated animals. Testes wet weight was reduced, but no modifications were detected in vas deferens or seminal vesicles. Postnatal treatment did not alter offspring body weight, open-field activity, sexual behavior and organ weight as well as did not delay testes descent but reduced the time until vaginal opening. Hypothalamic serotonin (5-HT), dopamine (DA) and noradrenaline (NA) as well as DA and NA metabolites were not modified by both prenatal and postnatal treatments. Increased striatal 3,4 dihydroxyphenylacetic acid (DOPAC) levels were observed after prenatal and postnatal treatments, while only postnatal 5-HT levels were increased. We propose that the present results indicate that prenatal ATZ exposure can have long lasting organizational effects on reproductive behavior of male rats, while postnatal exposure to this drug did not alter mating behavior. In relation to female rats, prenatal and postnatal exposures to ATZ accelerated puberty but did not alter sexual behavior. Neurochemical data show that both treatments increased striatal dopaminergic system activity, suggesting a central ATZ effect after perinatal exposure. PMID- 11943514 TI - Methamphetamine-gonadal steroid hormonal interactions: effects upon acute toxicity and striatal dopamine concentrations. AB - Methamphetamine (MA)-related deaths and nigrostriatal dopaminergic (NSDA) neurotoxicity are greater in males. The exact basis for this gender difference is not known, but data, which show that estrogen (E) can function as a protectant of both the cardiovascular and NSDA systems, suggest an important role for gonadal steroids in modulating toxicity to this psychostimulant. In the present report, we examined the effects of treatment with the gonadal steroid hormones E and testosterone (T) upon MA-induced toxicity within intact and castrated female and male CD-1 mice. Treatment of intact males with E produced a severe acute toxicity to MA, with only 41% (7/17) males surviving at 24-h post-MA. This incidence of mortality was significantly different from that of nonhormonally treated mice receiving an identical regimen of MA [94% survival (16/17)]. None of the other treatment groups showed mortality rates, which differed significantly from the nonhormonally treated mice. Striatal dopamine (DA) concentrations of E-treated female mice (intact or castrated) were significantly greater than that of the nonhormonally treated mice, which failed to differ statistically among each other. In an attempt to understand some of the bases for the mortality rates in E treated intact males, body temperatures, heart rates and heart catecholamine concentrations were measured from an additional group of intact male mice treated or not treated with E. Heart rates of E-treated intact males were significantly decreased compared with non-E-treated males. No statistically significant differences were obtained for body temperatures or heart catecholamine concentrations. These data demonstrate that MA induces an exacting, acute toxicity, which is specific for E-treated intact male mice and is associated with a reduction in heart rate. In addition, E can function as a neuroprotectant of NSDA system within female, but not male, mice. These data suggest that acute MA toxicity observed with E in intact male mice may result from a change in cardiac function. Accordingly, gonadal steroid hormones can function as critical modulators of both central and peripheral toxicological effects of MA. PMID- 11943515 TI - A preliminary investigation of the effects of maternal ethanol intake during gestation and lactation on brain adenosine A(1) receptor expression in rat offspring. AB - Ethanol exposure during fetal development can result in behavioral and neurological deficits, including reduced cognitive functions, retarded growth, and craniofacial abnormalities. Adenosine is an endogenous neuromodulator that fine-tunes the release and/or synaptic activities of several neurotransmitters, including glutamate, dopamine, and serotonin. Our aim was to determine whether ethanol exposure during early development affects adenosine receptors, particularly the A1 receptor subtype, in adult rats. Female rats were given water or 15% (vol/vol) ethanol in water prior to mating and throughout gestation and lactation. Sixty-day-old male rat offspring from these dams were randomly selected and assayed for adenosine A1 receptor expression in four brain areas: cortex, cerebellum, hippocampus, and striatum. Our results indicate that ethanol intake by dams decreased body and brain weights of offspring and reduced both A1 receptor mRNA and protein density in cortex and cerebellum. These preliminary findings indicate that ethanol intake by dams during pregnancy and lactation can affect adenosine A1 receptor signalling in the offspring. A pair-fed controlled study is warranted to explore these findings further. PMID- 11943516 TI - Could nitric oxide be an important mediator in opioid tolerance and morphine side effects? PMID- 11943517 TI - Double-blind evaluation of transdermal nitroglycerine as adjuvant to oral morphine for cancer pain management. AB - STUDY OBJECTIVES: To examine analgesia and adverse effects following transdermal application of nitroglycerine (a nitric oxide generator) combined with oral morphine, in cancer pain patients. DESIGN: Randomized, double-blind study. SETTING: Teaching hospital. PATIENTS: 36 patients suffering from cancer pain. INTERVENTIONS: Patients were divided into two groups (n = 18). All patients were regularly taking oral amitriptyline 50 mg at bedtime. Pain was evaluated using a 10-cm visual analog scale (VAS). The morphine regimen was individually adjusted to a maximal oral dose of 80 to 90 mg/day, to maintain the VAS score less than 4/10 cm. When patients complained of pain (VAS equal or greater than 4/10), despite taking 80 to 90 mg of oral morphine daily, the transdermal test drug was supplemented as follows: the control group received a placebo patch daily, and the nitroglycerine group received a 5-mg/24-hour nitroglycerine patch daily. Patients were free to manipulate their daily morphine consumption at the time the test drug was administered, to keep VAS less than 4/10 cm. After the introduction of the transdermal test drug, patients were evaluated by the staff on a weekly basis as outpatients, over four consecutive weeks. MEASUREMENTS AND MAIN RESULTS: The groups were similar in respect to demographic data and VAS pain scores before the treatment. The daily consumption of oral morphine was smaller in the nitroglycerine group compared with the control group after the 14th day of evaluation (p < 0.002). Patients from the control group in general complained of somnolence, compared with the nitroglycerine group. CONCLUSION: Transdermal nitroglycerine was an effective coadjuvant analgesic. In conjunction with its opioid tolerance sparing function, delivery of nitric oxide donors together with opioids may be of significant benefit in cancer pain management in delaying morphine tolerance and decreasing the incidence of adverse effects related to high doses of opioids. PMID- 11943518 TI - Intravenous regional anesthesia with clonidine in the management of complex regional pain syndrome of the knee. AB - STUDY OBJECTIVE: To evaluate the safety and efficacy of administering intravenous regional anesthesia (IVRA) with clonidine in the management of complex regional pain syndrome (CRPS) of the knee. DESIGN: Prospective, non-blinded study. SETTING: University-affiliated pain management center. PATIENTS: 7 patients with the diagnosis of CRPS of the knee. The diagnosis of CRPS was made if a) patients had at least four of these symptoms: allodynia, hyperesthesia, edema, vasomotor changes, pain with a burning quality, sudomotor changes, joint stiffness, or temperature differences between extremities; b) patients had significant pain relief (>75%) after a lumbar sympathetic block. INTERVENTIONS: Each patient received IVRA with a solution containing clonidine 1 microg/kg in a total volume of 50 mL 0.5% lidocaine. IVRA clonidine (IVRA-C) was performed on each patient up to six times (maximum of once a week for six weeks). MEASUREMETNS: Pain was assessed using a verbal pain scale (VPS) between 0 and 10 before each IVRA-C treatment. Duration of pain relief was defined as the time during which the patients experienced no pain (VPS = 0). Continuous electrocardiogram (ECG) and pulse oximetry (SpO2) were monitored and blood pressure was recorded every 5 minutes for the first 60 minutes after tourniquet deflation. MAIN RESULTS: Five patients received complete pain relief with 4 to 6 IVRA-C blocks. The two remaining patients reached a therapeutic plateau from IVRA-C, but had persistent anatomic lesions contributing to the recurrence of their CRPS. No patient experienced hypotension (mean arterial pressure < or = 20% baseline), hypoxemia (SpO2 < or = 90%), bradycardia (heart rate < or =H 60 beats/min), or excessive sedation. CONCLUSIONS: IVRA-C is a useful treatment modality in the management of CRPS of the knee. Clonidine doses of 1 microg/kg appear to be well tolerated without significant side effects. PMID- 11943519 TI - Effects of adding midazolam on the postoperative epidural analgesia with two different doses of bupivacaine. AB - STUDY OBJECTIVE: To investigate the interaction of midazolam with different doses of bupivacaine, by comparing the analgesic, sedative, and amnesic effects of continuous epidural midazolam with two different doses of bupivacaine. DESIGN: Prospective, randomized study. SETTING: Operating room and intensive care units of a university hospital. PATIENTS: 100 ASA physical status I and II postgastrectomy patients (40-70 yrs) without any complications. INTERVENTIONS: Patients were divided into four groups (n = 25) and administered continuous thoracic epidural infusion (40 mL/12 hr) of the drugs via a balloon infuser. The contents of the infuser (40 mL) were bupivacaine 180 mg with midazolam 20 mg (HM group), 90 mg with midazolam 20 mg (LM group), 180 mg without midazolam (HC group), or 90 mg without midazolam (LC group). As a rescue medication, 50 mg indomethacin suppository was the first choice, then IM pentazocine 15 mg. MEASUREMENTS: Analgesia and sedation scores, blood pressure (BP), heart rate, respiratory rate, oxygen saturation, amnesia, the number of rescue medications, and time to the first rescue medication were monitored. The area under the curves (AUCs) of analgesia and sedation scores, and BP were calculated. MAIN RESULTS: In the HM group, significantly better analgesia and sedation were obtained and the number of rescue medications given was the lowest of the four groups. The time to the first rescue medication was longest in the HM group, followed by the LM group, then the HC, and, finally, LC groups. The numbers of the patients with amnesia were greater in the HM and LM groups than the HC and LC groups. Blood pressure decreased significantly in the HM group but no treatment was necessary. The AUCs of analgesia and sedation scores, and BP were lowest in the HM group. CONCLUSIONS: Adding midazolam increased not only analgesic but also sedative effect with increasing dose of bupivacaine in a postoperative continuous epidural administration. PMID- 11943520 TI - Respiratory acidosis prolongs, while alkalosis shortens, the duration and recovery time of vecuronium in humans. AB - STUDY OBJECTIVE: To determine the effects of respiratory acidosis and alkalosis by mechanical ventilation on the onset, duration, and recovery times of vecuronium. DESIGN: Randomized, prospective study. SETTING: Operating rooms in the Sapporo Medical University Hospital and Kitami Red Cross Hospital. PATIENTS: 90 ASA physical status I and II patients undergoing lower abdominal surgery. INTERVENTIONS: Patients were randomly allocated to one of three groups by arterial carbon dioxide tension level (PaCO2; mmHg) after induction: hyperventilation group (PaCO2 = 25-35), normoventilation group (PaCO2 = 35-45), and hypoventilation group (PaCO2 = 45-55). Anesthesia was maintained by spinal block with inhalation of 50% to 66% nitrous oxide in oxygen and intermittent intravenous administration of fentanyl and midazolam with tracheal intubation. MEASUREMENTS AND MAIN RESULTS: After vecuronium 0.08 mg/kg was given, onset, duration, and recovery time were measured by mechanomyography (Biometer Myograph 2,000, Odense, Denmark). There were significant differences in the duration and recovery time of vecuronium among the normoventilation group (12.7 +/- 3.3 min and 11.8 +/- 2.8 min, respectively), the hyperventilation group (10.6 +/- 3.5 min and 9.2 +/- 2.7 min, respectively; p < 0.01), and the hypoventilation group (14.4 +/- 3.1 min and 15.0 +/- 3.7 min, respectively; p < 0.01) (mean SD). The closest significant correlation in this study was observed between recovery time and arterial blood pH (r = 0.57; p < 0.05). CONCLUSION: In humans, duration and recovery times of vecuronium are prolonged in respiratory acidosis and shortened in respiratory alkalosis. PMID- 11943521 TI - Frequency of hypotension and bradycardia during general anesthesia, epidural anesthesia, or integrated epidural-general anesthesia for total hip replacement. AB - STUDY OBJECTIVE: To evaluate the frequency of hypotension and bradycardia during integrated epidural-general anesthesia as compared with general anesthesia or epidural anesthesia alone. DESIGN: Prospective, randomized, open, multicenter study. SETTING: Inpatient anesthesia at 7 University or Hospital Departments of anesthesia. PATIENTS: 210 ASA physical status I, II, and III patients undergoing elective total hip replacement. INTERVENTIONS: Using a balanced randomization method, each hospital enrolled 30 consecutive patients who received integrated epidural-general anesthesia, epidural anesthesia, or general anesthesia. MEASUREMENTS AND MAIN RESULTS: Occurrence of clinically relevant hypotension (systolic arterial blood pressure (BP) decrease >30% from baseline), or bradycardia (heart rate (HR) <45 bpm) requiring pharmacologic treatment were recorded, as well as routine cardiovascular parameters. Clinically relevant hypotension during induction of nerve block was reported in 13 patients receiving epidural block (18%) and 16 patients receiving epidural-general anesthesia (22%) (p = 0.67). Subsequently, 22 of the remaining 54 patients in the epidural-general anesthesia group (41%) developed hypotension after the induction of general anesthesia, as compared with 16 patients of the general anesthesia group (23%) (p = 0.049). No differences in HR or in frequency of bradycardia were observed in the three groups. CONCLUSIONS: The induction of general anesthesia in patients with an epidural block up to T10 increased the odds of developing clinically relevant hypotension as compared with those patients who received no epidural block, and was associated with a twofold increase of the odds of hypotension as compared with the use of epidural anesthesia alone. PMID- 11943522 TI - Anesthesia with ketamine, propofol, and fentanyl decreases the frequency of postoperative psychosis emergence and confusion in schizophrenic patients. AB - STUDY OBJECTIVE: To investigate whether total IV anesthesia with ketamine, propofol, and fentanyl affects the frequency of postoperative psychosis emergence or confusion in schizophrenic patients. DESIGN: Prospective, controlled study. SETTING: Hirosaki National Hospital and Hakodate Watanabe Hospital. PATIENTS: 76 ASA physical status I and II schizophrenic patients taking chronic antipsychotic drugs and schedule for orthopedic surgery of extremities. INTERVENTIONS: In Group A (n = 38) patients, anesthesia was maintained with sevoflurane, nitrous oxide, and fentanyl. In Group B (n = 38) patients, anesthesia was maintained with ketamine, propofol, and fentanyl. MEASUREMENTS AND MAIN RESULTS: The frequency of psychosis emergence or confusion (54%) in Group A during the first 48 hours after surgery was significantly higher than the 30% figure in Group B. CONCLUSION: Ketamine, when combined with propofol and fentanyl, is an appropriate anesthetic drug for schizophrenic patients. PMID- 11943523 TI - Clinical properties of levobupivacaine or racemic bupivacaine for sciatic nerve block. AB - STUDY OBJECTIVE: To compare the intraoperative and postoperative clinical properties of the sciatic nerve block performed with either 0.5% bupivacaine or 0.5% levobupivacaine for orthopedic foot procedures. DESIGN: Randomized, double blind study. SETTING: Inpatient unit of a university-affiliated hospital. PATIENTS: 30 ASA physical status I and II patients undergoing elective hallux valgus repair under regional anesthesia. INTERVENTIONS: After administering intravenous (IV) midazolam premedication (0.05 mg/kg), a femoral nerve block was performed with 15 mL of mepivacaine 2%. Patients were then randomly allocated to receive, in a double-blind fashion, a sciatic nerve block with 20 mL of either 0.5% bupivacaine (n = 15) or 0.5% levobupivacaine (n = 15). MEASUREMENTS AND MAIN RESULTS: An observer who was blinded to the study drug recorded the onset time, quality, and duration of the sciatic nerve block. Postoperative analgesia consisted of 100 mg IV ketoprofen every 8 hours, with the first administration given at the patient's request. Mean (+/-SEM) onset time of the sciatic nerve block was 35 +/- 5 minutes for bupivacaine and 31 +/- 6 minutes for levobupivacaine (p = not significant [NS]). The duration of motor and sensory blocks with bupivacaine was 761 +/- 112 minutes and 790 +/- 110 minutes, respectively, and 716 +/- 80 minutes and 814 +/- 73 minutes, respectively, with levobupivacaine (p = NS). The first pain medication was requested after 844 +/- 96 minutes with bupivacaine and 872 +/- 75 minutes after levobupivacaine (p = NS). No differences in the quality of nerve block and patient satisfaction were reported between the two groups. CONCLUSIONS: A dose of 20 mL of 0.5% levobupivacaine induces sciatic nerve block of similar onset, duration, and intensity as the block produced by the same volume and concentration of the racemic solution of bupivacaine. PMID- 11943524 TI - Tracheal intubation conditions and cardiovascular effects after modified rapid sequence induction with sevoflurane-rapacuronium versus propofol-rapacuronium. AB - STUDY OBJECTIVES: To compare intubation conditions and hemodynamic effects resulting from rapid-sequence induction of anesthesia with sevoflurane rapacuronium and propofol-rapacuronium. DESIGN: Randomized, blinded study. SETTING: Operating suites of a large university-affiliated medical center. PATIENTS: 40 ASA physical status I and II adult patients without airway abnormalities who were scheduled for elective surgery requiring endotracheal intubation. INTERVENTIONS: Patients were randomly allocated to receive either sevoflurane inhalational induction (Group 1) or propofol (2 mg/kg) intravenous induction (Group 2). Group 1 patients were coached on how to perform vital capacity breathing and the anesthesia machine was primed with sevoflurane 8%, N2O:O2 3.5:1.5 L/min. In both groups, when loss of consciousness was established, rapacuronium 1.5 mg/kg was administered. After 50 seconds, an anesthesiologist blinded to the study entered the room and attempted laryngoscopy and intubation. MEASUREMENTS: Intubation conditions were graded as excellent, good, poor, or impossible according to Good Clinical Research Practice (GCRP) criteria. Arterial blood pressure and heart rate changes accompanying both induction techniques were also monitored and recorded. MAIN RESULTS: All patients were successfully intubated within 60 seconds. Clinically acceptable intubating conditions (excellent or good scores) were obtained in 19 of 20 Group 1 patients and in 19 of 20 Group 2 patients. Moderate tachycardia was encountered in both groups and mild systolic hypotension in the Group 2 patients. There were no complications. CONCLUSIONS: Modified rapid-sequence inhalational induction using sevoflurane and rapacuronium produced clinically acceptable intubation conditions within 60 seconds of muscle relaxant administration. The intubation conditions were similar to those produced after intravenous propofol and rapacuronium. PMID- 11943525 TI - Continuous epidural, not intravenous, droperidol inhibits pruritus, nausea, and vomiting during epidural morphine analgesia. AB - PURPOSE: To investigate whether continuous epidural droperidol and intravenous (IV) intraoperative droperidol inhibit pruritus and postoperative nausea and vomiting (PONV) during epidural morphine analgesia. DESIGN: Randomized, double blinded, controlled study. SETTING: Metropolitan cancer center. PATIENTS: 120 ASA physical status I and II patients undergoing thoracic or abdominal surgery with general anesthesia combined with epidural anesthesia. INTERVENTIONS: Patients received an intraoperative epidural injection of 2 mg morphine hydrochloride, followed postoperatively by a continuous epidural infusion of morphine hydrochloride 4 mg/day for 4 days. Patients were randomly allocated to four groups: Group A = control group, Group B = intraoperative single IV injection of droperidol (2.5 mg), Group C = postoperative continuous epidural droperidol infusion (2.5 mg/day), and Group D = intraoperative IV injection of droperidol (2.5 mg) and postoperative continuous epidural droperidol infusion (2.5 mg/day). MEASUREMENTS AND MAIN RESULTS: The frequency and severity of pruritus and PONV in each group were evaluated during the postoperative period. Continuous epidural infusion of droperidol significantly reduced the frequency and severity of pruritus and PONV induced by epidural morphine without causing significant side effects. Intraoperative single IV injection of droperidol was effective for PONV (p < 0.05) but not for pruritus. CONCLUSION: Postoperative epidural droperidol infusion significantly decreased both the frequency and severity of pruritus and PONV during postoperative continuous epidural morphine analgesia. IV intraoperative droperidol significantly reduced the frequency and the severity of PONV but not pruritus. PMID- 11943526 TI - Implementing ACC/AHA guidelines for the preoperative management of patients with coronary artery disease scheduled for noncardiac surgery: effect on perioperative outcome. AB - STUDY OBJECTIVE: To review the new consensus guidelines for cardiac testing for the patient with cardiac disease scheduled for elective, noncardiac surgery, and their impact on cardiac functional testing. DESIGN: Retrospective chart review study. SETTING: Tertiary care medical center. PATIENTS: 181 patients scheduled for elective, major surgery who met American College of Cardiology/American Heart Association (ACC/AHA) criteria for a preoperative stress test. INTERVENTIONS: A variety of tests were ordered, including treadmill stress testing, persantine thallium imaging, dobutamine echocardiography, and exercise stress echocardiography. MEASUREMENTS: The numbers of and outcome of the stress tests and the cardiac outcome of the patients who underwent cardiac testing and surgery were recorded. MAIN RESULTS: Abnormal tests occurred in 27 patients. Two patients declined treatment, eight patients had primary medical management, and the remainder (17) had cardiac catheterization. Results included no lesion (2 patients), angioplasty (4 patients), angioplasty plus stenting (1 patient), coronary artery bypass grafting (CABG) (4 patients), and delineated lesions treated with medical optimization (6 patients). One patient had CABG and declined further surgery. One patient had myocardial infarction 6 months after surgery that was treated by medical management after cardiac catheterization. The other 23 patients had surgery without cardiac complication within 1 year of surgery. Only 15% (27/180) of the patients with indications for a stress test had a positive result. Even fewer patients had any alteration of the perioperative period. Despite this finding, cardiac morbidity was very low. CONCLUSIONS: The guidelines for stress test may be over-sensitive, and further prospective clinical studies are indicated. PMID- 11943527 TI - Apnea and bradypnea in patients receiving epidural bupivacaine-morphine for postoperative pain relief as assessed by a new monitoring method. AB - STUDY OBJECTIVE: To evaluate postoperative breathing patterns with a new monitoring device in patients given bupivacaine-morphine epidural analgesia. DESIGN: Open explorative study. SETTING: Inpatient anesthesia in a university hospital setting. PATIENTS: 15 ASA physical status I and II patients aged 28 to 87 years and scheduled for major abdominal surgery. INTERVENTIONS: All patients underwent abdominal surgery with epidural anesthesia combined with general anesthesia. Postoperatively, they continued with epidural analgesia consisting of bupivacaine and morphine. On the first postoperative night, the breathing pattern was studied with a new noninvasive monitoring device measuring respiratory frequency and apnea. Arterial blood gas analysis was performed in case of apnea or low respiratory frequency. MEASUREMENTS AND MAIN RESULTS: A total of 84 alarm events were registered in 11 patients. Twenty-one percent (18/84) of the alarms were associated with arterial carbon dioxide tension (PaCO2) levels greater than 48.8 mmHg. Three of the four patients with PaCO2 levels greater than 48.8 mmHg were older than 80 years of age. CONCLUSION: The tested noninvasive monitoring device may detect abnormal respiratory breathing patterns in patients at risk for respiratory depression during epidural analgesia with bupivacaine-morphine. PMID- 11943528 TI - Dose requirements and recovery profile of an infusion of cisatracurium during liver transplantation. AB - STUDY OBJECTIVE: To examine the dose requirements and recovery profile of an infusion of cisatracurium during liver transplantation. DESIGN: Open-label, descriptive study. SETTING: University hospital. PATIENTS: 6 ASA physical status III and IV patients with end-stage liver disease, undergoing liver transplantation. INTERVENTIONS: Neuromuscular transmission was monitored electromyographically. After recovery of T1/T0 to 10%, cisatracurium was infused at an initial rate of 1.5 microg/kg/min. The infusion rate was adjusted to maintain T1/T0 at 10%. At the end of surgery, spontaneous recovery from the neuromuscular block was awaited. MEASUREMENTS AND MAIN RESULTS: The infusion rate of cisatracurium was 1.6 +/- 0.4 microg/kg/min. Before the anhepatic phase, this rate was 1.5 +/- 0.4 microg/kg/min; during the anhepatic phase it was 1.7 +/- 0.5 microg/kg/min; and after reperfusion it was 1.9 +/- 0.4 microg/kg/min. There was a significant difference between the cisatracurium infusion rates before and after the anhepatic phase (p < 0.05). Following termination of the infusion, the time to 25% recovery of T1/T0 was 19.2 +/- 6.1 minutes, the recovery index (25% to 75%) was 28.8 +/- 7.0 minutes, and the time for the train-of-four (TOF) ratio to reach 0.7 was 50.2 +/- 7.1 minutes. The time for the TOF ratio to reach 0.9 was 61.4 +/- 6.6 minutes. There was no difference in body temperature or pH during the consecutive stages of transplantation. CONCLUSIONS: The infusion dose requirement for cisatracurium during liver transplantation tended to be higher than previously reported in healthy patients; recovery appeared prolonged. In continuous infusion of cisatracurium during liver transplantation, the tendency toward higher dose requirements, the protracted duration of infusion, the non Hofmann elimination and/or other pharmacokinetic changes during transplantation might influence recovery from the neuromuscular block. Potential temperature or pH change during surgery seemed irrelevant in explaining the delayed recovery. PMID- 11943529 TI - Clonidine as coadjuvant in eye surgery: comparison of peribulbar versus oral administration. AB - STUDY OBJECTIVE: To determine whether the administration of peribulbar or oral clonidine would enhance analgesia and anesthesia in ophthalmologic surgery. DESIGN: Randomized double-blind study. SETTING: Teaching hospital. PATIENTS: 60 ASA physical status I and II adult patients scheduled for unilateral ophthalmologic surgery with peribulbar block. INTERVENTIONS: Patients were assigned to one of 4 groups, and premedicated with oral 2 mL volume (clonidine or placebo). The peribulbar eye block consisted of local anesthetics plus 1 mL of the test drug. The control group (CG) received oral saline as premedication and peribulbar saline as the test drugs. The clonidine eye group (Clo-eye G) received oral saline and peribulbar 30 microg clonidine. The clonidine oral group (Clo oral G) received oral 150 microg clonidine and peribulbar saline. The clonidine eye+oral group (Clo eye+oral G) had oral 75 microg clonidine and peribulbar 15 microg clonidine. MEASUREMENTS AND MAIN RESULTS: Perioperative assessment included anesthesia, analgesia, blood cortisol; and adverse effects. The groups were demographically similar. The latency time to the onset of the peribulbar block was shorter in the Clo-eye G compared to the CG (p < 0.05). The CG presented higher blood pressure levels throughout surgery, compared to the others (p < 0.05). The time to first rescue analgesics was longer in all patients who received peribulbar clonidine compared to the CG (p < 0.05). Analgesic consumption was lesser in the Clo-eye G compared to the CG (p < 0.05). The blood cortisol level was higher during the intraoperative period in all groups (preoperative vs. intraoperative values) (p < 0.01). CONCLUSION: Despite the higher intraoperative blood cortisol levels, 30 microg peribulbar clonidine decreased the onset time to anesthesia, while 15 and 30 microg peribulbar clonidine prolonged the time to first rescue analgesics in patients under peribulbar block, without increasing the frequency of adverse effects. Conversely, oral administration of clonidine alone did not enhance anesthesia or analgesia following eye block, suggesting a local mechanism of action of clonidine. PMID- 11943530 TI - The effect of fentanyl on hemodynamic and bispectral index changes during anesthesia induction with propofol. AB - STUDY OBJECT: To investigate the changes in hemodynamics and hypnotic levels during propofol infusion and tracheal intubation with and without fentanyl. DESIGN: Randomized, double-blinded study. SETTING: Teaching hospital. PATIENTS: 40 ASA physical status I adult patients scheduled for elective surgery. INTERVENTIONS: Patients were anesthetized with either propofol (Group P; n = 20) or 2 microg/kg of fentanyl IV followed by propofol (Group PF; n = 20). Propofol was infused at 20 mg/kg/hr throughout the study, and tracheal intubation was performed 10 minutes after the start of propofol infusion. MEASUREMENT AND MAIN RESULTS: Bispectral index monitoring (BIS) progressively decreased to about 50 in both groups during infusion of propofol, but no difference was found between the two groups. After tracheal intubation, BIS significantly increased but remained below 60 in both groups. Hypertensive responses to intubation were fewer in Group PF than Group P. CONCLUSIONS: Propofol administration 20 mg/kg/hr for 10 minutes is suitable in suppressing arousal reactions to tracheal intubation, but the addition of fentanyl is required to blunt the hemodynamic responses. PMID- 11943531 TI - Difficult airway management in a patient with severe aortic stenosis, coronary artery disease, and heart failure. AB - We report a patient with aortic stenosis, coronary artery disease, and heart failure in whom awake fiberoptic intubation was performed because the patient's mouth opening was insufficient as a result of disc prolapse of the mandibular joint. We planned to perform this procedure during conscious sedation, spontaneous respiration, and with stable hemodynamic parameters (no tachycardia, hypertension, or hypotension). After oral premedication with dikaliumclorazepate, the patient received clonidine. Fiberoptic intubation was performed while the patient breathed spontaneously. Throughout the procedure, no adverse events occurred. Clonidine is a valuable adjunct for conscious sedation and can be used safely for sympathicolysis, even in patients with aortic stenosis. PMID- 11943532 TI - Airway complication following functional endoscopic sinus surgery. AB - Difficulty breathing after upper airway surgery requires immediate evaluation and treatment. We present a case of airway compromise after sinus surgery due to edema of the uvula. The patient was admitted for observation overnight and discharged the next day. A discussion of potential airway changes after sinonasal surgery is presented. PMID- 11943533 TI - An unusual problem during central venous cannulation by Seldinger technique. PMID- 11943534 TI - Dislocation of the temporomandibular joint due to forceful yawning during induction with propofol. PMID- 11943535 TI - A coactivator code for transcription. PMID- 11943536 TI - BSD: a novel domain in transcription factors and synapse-associated proteins. AB - This article describes a novel domain, BSD, that is present in basal transcription factors, synapse-associated proteins and several hypothetical proteins. It occurs in a variety of species ranging from primal protozoan to human. The BSD domain is characterized by three predicted alpha helices, which probably form a three-helical bundle, as well as by conserved tryptophan and phenylalanine residues, located at the C terminus of the domain. PMID- 11943537 TI - Sec61beta--a component of the archaeal protein secretory system. AB - Sec61p/SecYEG complexes mediate protein translocation across membranes and are present in both eukaryotes and bacteria. Whereas homologues of Sec61alpha/SecY and Sec61gamma/SecE exist in archaea, identification of the third component (Sec61beta or SecG) has remained elusive. Using PSI-BLAST, the archaeal counterpart of Sec61beta has been detected. With the identification of the Sec61beta motif, functions for a universal family of archaeal proteins can be predicted and the archaeal translocon system can be definitively detected. PMID- 11943538 TI - Classical and novel steroid actions: a unified but complex view. PMID- 11943539 TI - Some like it hot. PMID- 11943540 TI - A database of transcriptional start sites. PMID- 11943541 TI - An active cytokine with a virus-like structure. PMID- 11943542 TI - Antisense peptides that inhibit beta-amyloid aggregation. PMID- 11943544 TI - Ribosome structure: revisiting the connection between translational accuracy and unconventional decoding. AB - The ribosome is a molecular machine that converts genetic information in the form of RNA, into protein. Recent structural studies reveal a complex set of interactions between the ribosome and its ligands, mRNA and tRNA, that indicate ways in which the ribosome could avoid costly translational errors. Ribosomes must decode each successive codon accurately, and structural data provide a clear indication of how ribosomes limit recruitment of the wrong tRNA (sense errors). In a triplet-based genetic code there are three potential forward reading frames, only one of which encodes the correct protein. Errors in which the ribosome reads a codon out of the normal reading frame (frameshift errors) occur less frequently than sense errors, although it is not clear from structural data how these errors are avoided. Some mRNA sequences, termed programmed-frameshift sites, cause the ribosome to change reading frame. Based on recent work on these sites, this article proposes that the ribosome uses the structure of the codon-anticodon complex formed by the peptidyl-tRNA, especially its wobble interaction, to constrain the incoming aminoacyl-tRNA to the correct reading frame. PMID- 11943545 TI - New insights into the mechanism of virus-induced membrane fusion. AB - Infection by enveloped viruses requires fusion between the viral and cellular membranes, a process mediated by specific viral envelope glycoproteins. Information from studies with whole viruses, as well as protein dissection, has suggested that the fusion glycoprotein (F) from Paramyxoviridae, a family that includes major human pathogens, has two hydrophobic segments, termed fusion peptides. These peptides are directly responsible for the membrane fusion event. The recently determined three-dimensional structure of the pre-fusion conformation of the F protein supported these predictions and enabled the formulation of: (1) a detailed model for the initial interaction between F and the target membrane, (2) a new model for Paramyxovirus-induced membrane fusion that can be extended to other viral families, and (3) a novel strategy for developing better inhibitors of paramyxovirus infection. PMID- 11943546 TI - Biogenesis of the mitochondrial TOM complex. AB - The translocase at the outer membrane of mitochondria (TOM complex) mediates the initial steps of the import of preproteins into the organelle, which are essential for mitochondrial biogenesis and, therefore, for eukaryotic cell viability. The TOM complex is a multisubunit molecular machine with a dynamic structure. The biogenesis of TOM is of special interest because the complex is required for its own biogenesis. This article describes the mechanisms by which Tom components are targeted to the mitochondria and inserted into the outer membrane. The assembly of newly synthesized subunits into the functional TOM complex might occur via assembly intermediates that are in equilibrium with the mature complex. PMID- 11943547 TI - Redox reactions of regulatory proteins: do kinetics promote specificity? AB - Signaling by redox state regulates the transcriptional and post-transcriptional events that control gene expression. To elucidate redox signaling in vivo, the effects of the reductive intracellular redox environment on regulatory redox events must be taken into account. This article focuses on proteins that contain regulatory disulfides, considering whether regulatory proteins can be oxidized and how the redox state of regulatory proteins can be uniquely controlled to allow redox signaling via specific pathways. It is possible that the favored kinetics of the redox reactions of regulatory proteins are important for attaining specificity in redox signaling. PMID- 11943548 TI - Protein-water interactions in a dynamic world. AB - Protein-water interactions are key to biological function. They have an underlying dynamic component that pervades the functional roles associated both with particular systems and with the properties of proteins in general. This article focuses on the specific ways in which the dynamics of water are important to protein structure, motion and adaptability to changes in the protein environment. PMID- 11943549 TI - Peter Mitchell and the ox phos wars. PMID- 11943550 TI - A biologist's guide to synchrotron facilities: the BioSync web resource. AB - Research at synchrotron radiation facilities, once the domain of high energy physics, now has a major impact on fields as diverse as immunology, neurobiology, physiology, molecular biology, medicine and biotechnology. This article describes the development of a comprehensive synchrotron portal and informational website (http://www.biosync.sdsc.edu) for biologists engaged in research at synchrotrons. The site automatically provides timely and accurate information in a unified format by gathering technical descriptions of synchrotron beamlines using modern information management practices. PMID- 11943553 TI - Too much of a good thing? PMID- 11943554 TI - Searching for clues to premature aging. PMID- 11943555 TI - Starvation amidst plenty: PDKs and diabetes mellitus. PMID- 11943557 TI - Potential roles for endometrial inhibins, activins and follistatin during human embryo implantation and early pregnancy. AB - The human endometrium is a remarkably dynamic tissue, undergoing cycles of proliferation, differentiation and breakdown every 28 days. In preparation for embryo implantation, the endometrium differentiates or decidualizes, involving widespread morphological and functional differentiation of endometrial stromal cells. If pregnancy occurs, the decidua regulates trophoblast invasion and forms the maternal component of the placenta. Uterine remodeling has long been known to be regulated by the ovarian steroid hormones 17beta-estradiol and progesterone; however, only recently has the importance of paracrine factors in mediating the cellular and biochemical changes been recognized. Many growth factors and cytokines, such as inhibins and activins, whose expression is generally limited to developmental and pathological states, are produced by actively remodeling endometrial cells, and play crucial roles in regulating endometrial cell function. Here, we present evidence for integral roles for the inhibin and activin family in the paracrine regulation of endometrial receptivity, decidualization and implantation. PMID- 11943558 TI - BNP in hormone-guided treatment of heart failure. AB - The pharmacotherapy of heart failure has become complex. Angiotensin-converting enzyme inhibitors (or angiotensin II receptor blockers), beta-blockers, spironolactone, diuretics and digoxin can be prescribed concurrently. Endothelin antagonists and combined inhibitors of converting enzyme and neutral endopeptidase are under investigation. Optimal dosing will become increasingly difficult to judge. Plasma brain natriuretic peptide (BNP) indicates the severity of left ventricular dysfunction. The C-terminal bioactive peptide and N-terminal BNP (N-BNP) circulate at concentrations related to cardiac status. We proposed that plasma levels of N-BNP would provide an index to guide drug treatment in established heart failure. Sixty-nine patients were randomized to treatment adjusted according to clinical criteria or plasma N-BNP. Hormone-guided therapy resulted in fewer clinical end points than did clinical management. This encourages further exploration of hormone guidance of anti-heart failure therapy, which could be extended to patients with preserved ejection fraction, in addition to those with established systolic dysfunction. PMID- 11943559 TI - Genetic analysis of insulin signaling in Drosophila. AB - Studies in the fruit fly Drosophila melanogaster and the nematode Caenorhabditis elegans have revealed that components of the insulin signaling pathway have been highly conserved during evolution. Genetic analysis in Drosophila suggests that structural conservation also extends to the functional level. Flies carrying mutations that reduce insulin signaling have a growth deficiency phenotype similar to that seen in mice with disruptions of genes encoding insulin-like growth factors (IGFs) or the IGF-I receptor. Recent studies in flies have demonstrated a role for the insulin signaling pathway in the regulation of metabolism, reproduction and lifespan via modulation of central neuroendocrine pathways. Similarly, mice with loss of brain insulin receptors or insulin receptor substrate 2 deficiency exhibit neuroendocrine defects and female infertility. These parallels suggest that the insulin system has multiple conserved roles, acting directly to modulate growth and indirectly, via the neuroendocrine system, to modulate peripheral physiology in response to changes in nutrient availability. PMID- 11943560 TI - Prostate phenotypes in estrogen-modulated transgenic mice. AB - Estrogen-modulated transgenic mice, such as estrogen receptor-knockouts (alphaERKO and betaERKO), aromatase-knockout (ArKO) and aromatase-overexpressing (AROM+) mice, have contributed to our understanding of the roles of estrogens in male reproductive biology, including prostate growth and development. Varying pathological changes of the prostate have been described as being the result of aberrant actions of estrogen, both directly through the estrogen receptors or indirectly by altering the endocrine status of these mice. This article identifies the consequences of aberrant estrogen signaling on prostate growth and development. Further characterization and manipulation of these estrogen modulated transgenic mice will lead to a more complete understanding of the hormonal regulation of the mammalian prostate gland. PMID- 11943561 TI - Genetic control of early folliculogenesis in mice. AB - Perinatally, granulosa cells encase individual oocytes within the ovary to form primordial follicles. The initial stages of folliculogenesis are independent of gonadotropins and involve cell-autonomous and non-cell-autonomous factors. Although still poorly understood at a molecular level, successful follicle formation and initiation of follicle growth must involve genetic networks both in germ and in somatic cells. Mouse models offer useful windows into these essential processes. By investigating phenotypes of mouse lines lacking specific gene products, genetic hierarchies that regulate the initial stages of folliculogenesis are being elucidated. These investigations will provide insight into the regulation of mammalian fertility. PMID- 11943568 TI - Gender differences in stress and immune function in adolescents. Proceedings of a workshop. May 7, 1999, Stanford, California, USA. PMID- 11943562 TI - Lipases and HDL metabolism. AB - Plasma levels of high-density lipoprotein (HDL) cholesterol are strongly inversely associated with atherosclerotic cardiovascular disease, and overexpression of HDL proteins, such as apolipoprotein A-I in animals, reduces progression and even induces regression of atherosclerosis. Therefore, HDL metabolism is recognized as a potential target for therapeutic intervention of atherosclerotic vascular diseases. The antiatherogenic properties of HDL include promotion of cellular cholesterol efflux and reverse cholesterol transport, as well as antioxidant, anti-inflammatory and anticoagulant properties. The molecular regulation of HDL metabolism is not fully understood, but it is influenced by several extracellular lipases. Here, we focus on new developments and insights into the role of secreted lipases on HDL metabolism and their relationship to atherosclerosis. PMID- 11943569 TI - Gender differences in emotional responses to interpersonal stress during adolescence. PMID- 11943570 TI - Boys' and girls' responses to stress: affect and heart rate during a speech task. AB - PURPOSE: To examine gender differences in heart rate and reports of negative and positive affect among adolescents during a speech task. METHODS: Subjects were 133 adolescents, 73 girls and 60 boys, ages 14-18 years. Subjects were randomly assigned to speak for 10 minutes and asked either to free associate or to describe the most stressful event in their lives. Their heart rate was assessed at baseline and at 5 and 10 minutes after the task began. Self-ratings of negative and positive affect were assessed at baseline and at 10 minutes after the beginning of the task. Data were analyzed by Student's t-tests for independent samples, analyses of covariance, and Spearman rank-order correlation coefficients. RESULTS: Girls rated their negative affect significantly higher at baseline in comparison to boys. In addition, girls had a higher baseline heart rate. However, boys' increase in heart rate 5 minutes into the speech task was significantly greater than that of girls. Also, the free association task elicited significantly greater increases in heart rate compared to a task in which the adolescent described his or her most stressful life event. However, the stressful event task elicited greater negative and less positive affect. Significant relationships between change in heart rate and ratings of post-task affect were found among girls but not among boys, suggesting that the girls' ratings of their affect are more congruent with their physiological reactions to a stressful speech task. CONCLUSIONS: These results suggest that gender and task differences must be considered in assessing adolescents' stress. PMID- 11943571 TI - Life stressors, social resources, and coping skills in youth: applications to adolescents with chronic disorders. AB - OVERVIEW: After setting out a conceptual framework that focuses on how personal and social resources aid adolescents in managing acute and chronic stressors, I describe methods by which to assess adolescents' family environments and specific life stressors and social resources, and the approach and avoidance coping responses adolescents use to manage life stressors. I then review some research that employs these concepts and methods to focus on the families and life contexts, and coping skills, of youth with chronic medical disorders, including juvenile rheumatic disease (JRD). I close by drawing implications for assessment and intervention and describing some fruitful areas for future research, such as examining the reciprocal linkages between parental and youth behavior, how adolescents' personal characteristics shape their life context, and how life crises and transitions enhance adolescents' development and maturation. PMID- 11943572 TI - Stress in adolescent females: relationship to autoimmune diseases. PMID- 11943573 TI - Autoimmunity, dehydroepiandrosterone (DHEA), and stress. PMID- 11943574 TI - Gender differences in puberty. PMID- 11943575 TI - Puberty and the emergence of gender differences in psychopathology. PMID- 11943576 TI - Gender differences in the development and function of the immune system. PMID- 11943577 TI - Can stress participate in the pathogenesis of autoimmune disease? PMID- 11943578 TI - Expression of autoimmunity in the transition from childhood to adulthood: role of cytokines and gender. PMID- 11943579 TI - Liminality and breastfeeding: women negotiating space and two bodies. AB - It is almost universally accepted that breastfeeding infants is nutritionally superior to bottle-feeding. However, despite this medical advice, in many countries breastfeeding rates remain low and in the UK, rates are relatively static. The literature on breastfeeding has discussed international rates and the broad socio-economic factors influencing these rates. Through an observational study of a group of breastfeeding and non-breastfeeding women in the United Kingdom, this research utilises contemporary theoretical perspectives on the body, space and rites of passage, and investigates the reasons why some breastfeeding mothers may be in a liminal period, and the breastfeeding event itself, at times, a liminal and marginalised act. The paper argues that, for the group studied, breastfeeding is sometimes discouraged by its medicalisation, and that breastmilk and breastfeeding are often considered by mothers to be embarrassing. Many of the women studied regarded certain public and private places to be unacceptable places to breastfeed and claimed to modify their behaviour accordingly. The paper demonstrates the value of conducting locally based qualitative research into breastfeeding experiences, and of using theoretical perspectives from post-medical geography to interpret women's experiences. PMID- 11943580 TI - Teen births, income inequality, and social capital: developing an understanding of the causal pathway. AB - Many studies have demonstrated a relationship between income inequality and poor health, but how does income inequality impact health? One possible explanation is that greater income inequality undermines social capital (social cohesion, civic engagement, and mutual trust in a community). We conducted path analyses of the relationship between income inequality, poverty, and teen birth rate, testing for the mediating effect of social capital in 39 US states. Birth rate was affected by both poverty and income inequality, though income inequality appeared to affect teen birth rate primarily through its impact on social capital. PMID- 11943581 TI - Spatial filtering using a raster geographic information system: methods for scaling health and environmental data. AB - Despite the use of geographic information systems (GIS) in academic research, it is still uncommon for public health officials to use such tools for addressing health and environmental issues. Complexities in methodological issues for addressing relationships between health and environment, investigating spatial variation of disease, and addressing spatial demand and supply of health care service, hinder the use of GIS in the health sector. This paper demonstrates simple spatial filtering methods for analyzing health and environmental data using a raster GIS. Computing spatial moving average rates reduces individual affects and creates a continuous surface of phenomena. Another spatial analytical method discussed is computation of exposure status surfaces including neighbors' influences weighted by distance decay. These methods describe how health and environmental data can be scaled in order to better address health problems. Spatial filtering methods are demonstrated using health and population surveillance data within a GIS that were collected for approximately 210,000 people in Matlab, Bangladesh. PMID- 11943582 TI - Geography of child mortality clustering within African families. AB - After decades of sustained child mortality reductions, infant and early childhood mortality levels in Africa remain high. This can partly be ascribed to the concentration of child mortality within particular families, communities or geographic localities. Strong mortality clustering is indicative of marked social inequality and of an unequal distribution of health and health-related resources and infrastructures. It also signifies a concentration of nutritional and sanitary behaviors harmful to the good health and longevity of children. Finally, it likely points to the existence of particular genetic problems in certain families, or environmental problems within specific communities. Using nationally representative family level data from all sub-regions of Africa, two important findings emerge from this study. First, levels of mortality have generally declined in all countries over time, and as mortality decreases, mortality clustering tends to follow the same trend. Second, bio-demographic covariates have a more important effect on familial mortality clustering risks than socio economic ones. PMID- 11943583 TI - Health care reform in New Zealand: the diversity of gender experience. AB - Women are a heterogeneous group with varying experiences and needs of health care systems. It is important that we recognise not only differences between women, but also that individual women may have different, even contradictory, health care issues and needs. These may vary according to women's different roles, identities, contexts, and resources (financial, social, etc.). This paper explores situated ideas about identity, gender, and place and how these relate to perceptions of accessibility of health care services in the context of New Zealand's restructured health care system. PMID- 11943584 TI - Urban air pollution and health: an ecological study of chronic rhinosinusitis in Cologne, Germany. AB - The study investigates the association between outdoor air pollution and chronic rhinosinusitis (CRS) in Cologne, Germany. First, using their addresses, all patients treated for CRS at the ENT Department of the University Hospital between 1990 and 1999 were assigned to one of the 85 city districts. Second, indicators of air pollution (SO(2), NO(x), TSP) were linked to these areas. Third, to control for socioeconomic confounding, data reflecting the socioeconomic and demographic composition of these districts were collected. Regression analyses reveal weak but consistent statistical effects of pollution on the prevalence of CRS in those parts of the city with air pollution levels above average. PMID- 11943585 TI - An ecological study of the relationship between social and environmental determinants of obesity. AB - There is growing concern with the increasing prevalence of obesity in industrialised countries, a trend that is more apparent in the poor than in the rich. In an ecological study, the relationship between an area measure of socioeconomic status (SES) and the density of fast-food outlets was examined as one possible explanation for the phenomenon. It was found that there was a dose response between SES and the density of fast-food outlets, with people living in areas from the poorest SES category having 2.5 times the exposure to outlets than people in the wealthiest category. The findings are discussed. PMID- 11943586 TI - LEKTI: a multidomain serine proteinase inhibitor with pathophysiological relevance. AB - Proteinase inhibitors are important negative regulators of proteinase action in vivo and are thus involved in several pathophysiological processes. Starting with the isolation of two new peptides from human blood filtrate, we succeeded in cloning a cDNA encoding the precursor protein for a novel 15-domain Kazal-type related serine proteinase inhibitor. Two of the 15 domains almost exactly match the Kazal-type pattern, whereas the other 13 domains exhibit only four instead of six cysteine residues. Since the corresponding gene is expressed in several lympho-epithelial tissues, we termed this inhibitor lympho-epithelial Kazal-type related inhibitor (LEKTI). For three of the 15 LEKTI domains, we demonstrated a significant trypsin-inhibiting activity. Recent results of another group show a relation between mutations within the LEKTI gene and the severe congenital disorder Netherton syndrome. In this review article, we give an overview of the already known data on the structure, processing, gene expression, and pathophysiological role of LEKTI. PMID- 11943587 TI - Getting to know protein kinase D. AB - The protein kinase D (PKD) enzymes represent a new family of second messenger stimulated kinases, with diacylglycerol as a prime, but not the sole, mediator of activation. Their molecular architecture features a catalytic domain, unrelated to that of all PKC family members, and a large inhibitory, regulatory domain, comprised of two Zinc fingers, and a pleckstrin homology domain. These different sub-domains play distinctive roles in the activation, translocation and biological functions of the kinase. The enzymes have been implicated in signalling mechanisms controlling cell proliferation and programmed cell death and in metastasis, immune responses, and Golgi restructuring and function. A variety of proteins specifically interact with the different sub-domains of the enzymes and direct their wide range of cellular functions. PMID- 11943588 TI - BRCA1-dependent and independent functions of BARD1. AB - Breast cancer susceptibility gene 1 (BRCA1)-associated RING domain (BARD1) was discovered as a protein interacting with the RING domain of BRCA1. It is structurally homologous to BRCA1 with which it shares the conserved RING finger and BRCT domains. BARD1 is strongly expressed in spleen and testis, correlated with the expression of BRCA1. Co-localization of BARD1 with BRCA1 and other repair proteins indicate a function in DNA repair. A potential role of BARD1 BRCA1 complexes in ubiquitination of RNA Pol II, and the interaction of BARD1 with polyadenylation factor CstF-50, thus inhibiting mRNA processing, provide mechanisms for tumor suppression. BRCA1-independent functions of BARD1 were first noted by its inordinate expression in hormonally regulated uterine tissue. BARD1 repression lead to a premalignant phenotype in mammary gland epithelial cells. The interaction of BARD1 with NF-kappaB, suggests modulation of transcriptional activity independent of BRCA1. Elevated BARD1 expression in apoptotic tumor cells was found associated with anti-BARD1 immune response thus leading to new therapeutic approaches. PMID- 11943589 TI - Corneal stromal cells (keratocytes) express thrombospondins 2 and 3 in wound repair phenotype. AB - Members of the thrombospondin (TSP) family of proteins have been implicated in wound healing. The cells of the corneal stroma (keratocytes) are capable of synthesising TSP-1 in a wound repair phenotype, but do not appear to produce the protein in the normal human adult cornea. We employed reverse-transcriptase polymerase chain reaction (RT-PCR) to determine whether human corneal stromal cells can express TSPs other than TSP-1. Cultured keratocytes contained messenger RNA (mRNA) for TSP-2 and TSP-3 (in addition to TSP-1), but not for TSP-4 or cartilage oligomeric matrix protein (COMP; TSP-5). Keratocytes in the normal cornea contained mRNA for TSP-1 but not for other TSPs. The distribution of keratocyte TSP-2 and TSP-3 immunoreactivity had some similarities to that of TSP 1 and, like TSP-1, neither protein could be detected in the cells of the normal corneal stroma. The observations suggest that keratocytes in wound repair phenotype produce TSP-2 and TSP-3 in addition to TSP-1. TSPs may play a pivotal role in corneal stromal repair and, since TSP-1 and TSP-2 have anti-angiogenic properties, may also have a function in regulating the avascularity of the central cornea. PMID- 11943590 TI - Urinary desmosine excretion is inversely correlated with the extent of emphysema in patients with chronic obstructive pulmonary disease. AB - An enhanced proteolysis of lung interstitium is key event in the pathogenesis of emphysema, a major constituent of chronic obstructive pulmonary disease. To assess whether urinary desmosine and/or hydroxyproline may be used as a marker of lung destruction we studied urinary excretions of these products in 20 patients with chronic obstructive pulmonary disease and in 19 appropriate controls in 24h urine collection samples. For desmosine measurements, we developed a new indirect competitive enzyme-linked immunosorbent assay. The extent of emphysema was measured in high resolution computed tomography (CT) scans, by considering lung area with CT numbers <-950 Hounsfield units (HU). Urinary desmosine excretion was significantly higher in patients with chronic obstructive pulmonary disease than in controls (294+/-121 microg versus 183+/-93 microg, P=0.003), and was unrelated with both age and smoking habits. In patients with no evidence or only mild emphysema, desmosine excretion values were significantly higher (P=0.006) than those of patients with moderate to severe emphysema. In patients with chronic obstructive pulmonary disease, urinary hydroxyproline excretion was positively correlated with urinary desmosine excretion but on the average, it was not different from that of controls. These data indicate that urinary desmosine is a sensitive biological marker of lung elastin catabolism. The relatively low levels of urinary desmosine observed in patients with severe emphysema may be accounted for a decrease in elastin catabolism due to reduced lung elastin mass. Urinary desmosine may be used to identify subjects at risk of developing emphysema and to assess the efficacy of therapeutic interventions. PMID- 11943592 TI - The purification and characterization of mu-calpain and calpastatin from ostrich brain. AB - Calcium-activated neutral proteinases (CANPs) and their endogenous specific inhibitor calpastatin are found in a wide variety of vertebrate and invertebrate tissues. The CANPs are cysteine proteinases that have an absolute requirement for Ca(2+) for activity. mu-Calpain and calpastatin were purified by successive chromatographic steps on Toyopearl-Super Q 650S and Pharmacia Mono Q HR 5/5 columns. The enzyme has a M(r) of 84KDa using sodium dodecyl sulphate polyacrylamide gel electrophoresis (SDS-PAGE), a M(min) of 79KDa from amino acid analysis and an pI of 5.2. Calpastatin has a M(r) of 323KDa using denaturing gradient PAGE and a pI of 4.7. The amino acid composition of mu-calpain revealed 689 residues and the pH and temperature optima were found to be 7.5 and 37 degrees C, respectively. mu-Calpain underwent a Ca(2+)-dependent autoproteolysis producing a fragment of 82KDa. The N-terminal sequence of mu-calpain showed 24 and 18% sequence identity with human and bovine mu-calpain. PMID- 11943591 TI - Fatty acid profiles and lipid peroxidation of microsomes and mitochondria from liver, heart and brain of Cairina moschata. AB - Studies were done to analyze the fatty acid composition and sensitivity to lipid peroxidation (LP) of mitochondria and microsomes from duck liver, heart and brain. The fatty acid composition of mitochondria and microsomes was tissue dependent. In particular, arachidonic acid comprised 17.39+/-2.32, 11.75+/-3.25 and 9.70+/-0.40% of the total fatty acids in heart, liver and brain mitochondria respectively but only 13.39+/-1.31, 8.22+/-2.43 and 6.44+/-0.22% of the total fatty acids in heart, liver and brain microsomes, respectively. Docosahexahenoic acid comprised 17.02+/-0.78, 4.47+/-1.02 and 0.89+/-0.07% of the total fatty acids in brain, liver and heart mitochondria respectively but only 7.76+/-0.53, 3.27+/-0.73 and 1.97+/-0.38% of the total fatty acids in brain, liver and heart microsomes. Incubation of organelles with ascorbate-Fe(2+) at 37 degrees C caused a stimulation of LP as indicated by the increase in light emission: chemiluminescence (CL) and the decrease of arachidonic acid to: 5.17+/-1.34, 8.86+/-0.71 and 5.86+/-0.68% of the total fatty acids in heart, liver and brain mitochondria, respectively, and to 4.10+/-0.61 in liver microsomes. After LP docosahexahenoic acid decrease to 7.29+/-1.47, 1.36+/-0.18 and 0.30+/-0.11% of the total fatty acids in brain, liver and heart mitochondria. Statistically significant differences in the percent of both peroxidable fatty acids (arachidonic and docosahexaenoic acid) were not observed in heart and brain microsomes and this was coincident with absence of stimulation of LP. The results indicate a close relationship between tissue sensitivity to LP in vitro and long chain polyunsaturated fatty acid concentration. Nevertheless, any oxidative stress in vitro caused by ascorbate-Fe(2+) at 37 degrees C seems to avoid degradation of arachidonic and docosahexaenoic acids in duck liver and brain microsomes. It is possible that because of the important physiological functions of arachidonic and docosahexaenoic acids in these tissues, they are protected to maintain membrane content during oxidative stress. PMID- 11943593 TI - Solenin, a novel protein with translation-inhibiting activity from the traditional Chinese medicinal fish, the sea dragon Solenognathus hardwickii. AB - A single-chained protein designated solenin was isolated from Solenognathus hardwickii, a fish used as traditional Chinese medicinal material. Solenin was capable of inhibiting translation in a cell-free rabbit reticulocyte lysate system with an IC(50) of 2 microM and expressing a ribonuclease activity of 0.8U/mg toward yeast transfer RNA, but it lacked N-glycosidase activity characteristic of ribosome inactivating proteins Solenin exhibited a molecular weight of 18kDa and possessed an N-terminal sequence AHDAEVNEVKAQVAA. The protein was adsorbed on three types of chromatographic media: Affi-gel blue gel, CM Sepharose and Mono S. It was devoid of antifungal and lectin activities. PMID- 11943594 TI - Chromatin structure analysis of the rat Na, K-ATPase beta2 gene 5'-flanking region. AB - The Na, K-ATPase is formed by two major subunits (alpha and beta) encoded by a gene family of at least four alpha and three beta isoforms. These genes show distinctive expression patterns involving complex tissue-specific and developmental regulation, although the control mechanisms are not well understood. Here we study the role of chromatin structure in the tissue-specific expression of rat Na, K-ATPase beta2 isoform, which is mainly found in the central nervous system. We have examined the presence and characteristics of nuclease hypersensitive sites and the cytosine methylation patterns in the 5' flanking region of the beta2 isoform gene from various nuclear preparations. Our results show that in this 5'-flanking region there is only one nuclease hypersensitive site. It is located upstream of the transcription initiation site and shows tissue-specific characteristics. Digestion with deoxyribonuclease I (DNase I), S1 nuclease and micrococcal nuclease yield patterns consistent with a triple-helix structure present only in the active state of the promoter. We also demonstrate that the 5'-flanking region of the beta2 gene co-localizes with a CpG island free of methylation in every tissue tested. The results presented here support a role for specific chromatin remodeling events in the regulation of the Na, K-ATPase beta2 gene expression. They also provide the basis for future studies of the transcription factors involved in the regulation of this gene. PMID- 11943595 TI - Urea-induced unfolding studies of free- and ligand-bound tetrameric ATP-dependent Saccharomyces cerevisiae phosphoenolpyruvate carboxykinase. Influence of quaternary structure on protein conformational stability. AB - ATP-dependent phosphoenolpyruvate (PEP) carboxykinases are found in plants and microorganisms, and catalyse the reversible formation of PEP, ADP, and CO(2) from oxaloacetate plus ATP. These enzymes vary in quaternary structure although there is significant sequence identity among the proteins isolated from different sources. To help understand the influence of quaternary structure in protein stability, the urea-induced unfolding of free- and substrate-bound tetrameric Saccharomyces cerevisiae PEP carboxykinase is described and compared with the unfolding characteristics of the monomeric Escherichia coli enzyme [Eur. J. Biochem. 255 (1998) 439]. The urea-induced denaturation of S. cerevisiae PEP carboxykinase was studied by monitoring the enzyme activity, intrinsic protein fluorescence, circular dichroism (CD) spectra, and 1-anilino-8 naphthalenesulfonate (ANS) binding. The unfolding profiles were multi-steps, and formation of hydrophobic structures were detected. The data indicate that unfolding and dissociation of the enzyme tetramer are simultaneous events. Ligand binding, most notably PEP in the presence of MnCl(2), conferred a marked protection against urea-induced denaturation. A similar protection effect was found when N-iodoacetyl-N'-(5-sulfo-1-napthyl)ethylene diamine (1,5-I-AEDANS) was covalently bound at Cys(365), within the active site region. Refolding experiments indicated that total recovery of tertiary structure was only obtained from samples previously unfolded to less than 30%. In the presence of substrates, complete refolding was achieved from samples originally denatured up to 50%. The unfolding behaviour of S. cerevisiae PEP carboxykinase was found to be similar to that of E. coli PEP carboxykinase, however all steps take place at lower urea concentrations. These findings show that, at least for monomeric and tetrameric ATP-dependent PEP carboxykinases, quaternary structure does not contribute to protein conformational stability. PMID- 11943596 TI - Converting troponin C into calmodulin: effects of mutations in the central helix and of changes in temperature. AB - Calmodulin (CaM) and troponin C (TnC) are the most similar members of EF-hand family and show few differences in the primary structure. Here, we use mutants of troponin that mimic calmodulin and changes in temperature to investigate the factors that determine their specificity as regulatory proteins. Using a double mutant of troponin that resembles calmodulin in lacking both the N-terminal helix and KGK(91-93) we observe a small difference from troponin in binding to the erythrocyte Ca(2+)-ATPase, and an improvement in enzyme activation. A triple mutant, where in addition, the residues 88-90 are replaced with the corresponding sequence from calmodulin is equivalent to calmodulin in maximal activation, and it restores protein ability to increase Ca(2+) affinity for the enzyme. However, this mutant also binds less tightly (1/100) than calmodulin. Remarkably, a decrease in temperature has a more marked effect in protein binding than either mutation, reducing the difference in affinities to 18-fold, but without any improvement in their ability to increase Ca(2+) affinity for the enzyme. Spectroscopic analysis of hydrophobic domain exposure in EF-hand proteins was carried out using 8-anilino-1-naphthalenesulfonic acid (ANS). The probe shows a much higher fluorescence when bound to the complex Ca(4)-calmodulin than to Ca(4) troponin. Decreasing the temperature exposes additional hydrophobic regions of troponin. Changing the Mg(2+) concentration does not affect their bindings to the enzyme. It is suggested that the requirements for troponin to mimic calmodulin in binding to the target enzyme, and those for activating it, are met by different regions of the protein. PMID- 11943597 TI - Mn(2+) ions reduce the enzymatic and pharmacological activities of bothropstoxin I, a myotoxic Lys49 phospholipase A(2) homologue from Bothrops jararacussu snake venom. AB - Bothropstoxin-I (BthTX-I), a myotoxic Lys49 phospholipase A(2) (PLA(2)) homologue isolated from Bothrops jararacussu snake venom, causes a range of biological effects, including myonecrosis, mouse paw edema, irreversible neuromuscular blockade and lysis of cell cultures. Among eight divalent cations assayed, Mn(2+) was the most effective in reducing mouse paw edema induced by BthTX-I (25 microg). Preincubating BthTX-I with Mn(2+) (1.0mM) reduced mouse paw edema by 70% and myotoxicity by 60% in mice injected i.m. with 50 microg toxin. Mn(2+) (50 microl of a 1mM solution) administered within 1min at the site of toxin injection was still but less effective in antagonising BthTX-I-induced myotoxicity. Mn(2+) (1.0mM) completely prevented BthTX-I (1.4 microM)-induced neuromuscular blockade in the mouse phrenic-nerve diaphragm preparation. Mn(2+) (0.25mM) protected about 85% of NB41A3 cells from lysis when coincubated with BthTX-I (1.0 microM) for 25h. Preincubation with 0.25mM Mn(2+) increased the sensitivity of the cells to subsequent lysis by BthTX-I in the absence of Mn(2+). BthTX-I (1 microM) caused extensive fatty acid release (from 0.8% of the total radiolabeled lipid in control cells to 56% with toxin) when incubated with the NB41A3 cell line for 25h. PLA(2) activity observed in cell cultures after addition of BthTX-I was considerably reduced by 0.25mM Mn(2+). Mn(2+) ions constitute a promising agent to assess the action mechanism and the effects of enzymatic inhibition on the pharmacological activity of Lys49 PLA(2) homologues. PMID- 11943598 TI - G(1)-phase specific apoptosis in liver carcinoma cell line induced by copper-1,10 phenanthroline. AB - Reactive oxygen species play an important role in the mediation of cell killing. But the mechanistic links between reactive oxygen species (ROS) and cell death remains unclear. There was a speculation that ROS, especially hydroxyl radicals can induce necrosis but not apoptosis in cells treated with copper-1,10 phenanthroline, IICu(OP)(2). In this paper, liver carcinoma cell line (Bel-7402) was treated with IICu(OP)(2) and its effect was examined by several means. Cells were found to undergo changes characteristic of apoptosis. Hoechst staining showed apoptotic body appeared in the cells induced by IICu(OP)(2). When DNA extracted from the cells treated with IICu(OP)(2) was analyzed by agarose gel electrophoresis it generated 'ladder' pattern of discontinuous DNA fragments. Sub G(1) peak was detected in treated cells. Furthermore, two different flow cytometric methods were used, each allowing us to relate the apoptotic cells to the position the cell-cycle position. Apoptosis induced by IICu(OP)(2) was limited to G(1)-phase cells. Using cyclin analysis, the expression of cyclin E in G(1) was blocked. Thus, it was concluded that IICu(OP)(2) can induce G(1)-phase specific apoptosis in Bel-7402. PMID- 11943599 TI - Induction of fascin spikes in breast cancer cells by activation of the insulin like growth factor-I receptor. AB - Insulin-like growth factor-I receptor (IGF-IR) signaling contributes to the formation of mammary carcinomas and has chiefly been studied with regard to the proliferative and anti-apoptotic effects of IGF-IR signaling. However, IGF-IR activation also affects the actin cytoskeleton and alterations in cell migratory behavior are of known importance for the malignant conversion and metastasis of epithelial cells. The actin-binding protein fascin is found in cell projections and spikes that are involved in the locomotion of mesenchymal cells. Fascin expression is typically low in normal epithelial cells, but is markedly upregulated in several types of carcinomas. Here, we also demonstrate increased fascin expression in breast carcinoma cell lines and adopt MCF-7 human mammary carcinoma cells that over-express wild-type or kinase-inactivated forms of the IGF-IR as a model system to test the hypothesis that IGF-IR activation induces fascin projections. We show that the time-dependent dissociation of cell colonies that occurs upon receptor activation by IGF-I involves the formation of dynamic, fascin-containing lateral cell projections that co-localize with ruffling membranes in association with protrusive activity and cell migratory phenotype. The molecular mechanism of these effects is completely dependent on IGF-IR tyrosine kinase activity and is mediated by a phosphatidylinositol (PI) 3-kinase dependent process. In demonstrating transduction of fascin spike assembly by activation of a peptide growth factor receptor, these novel data reveal a wide role for fascin spikes in cell motility and provide new insight into the complex effects of IGF-IR signaling on actin cytoskeletal organization. PMID- 11943600 TI - Increased expression level of the splicing variant of SIP1 in motor neuron diseases. AB - Survival motor neuron (SMN) interacting protein 1 (SIP1) interacts with SMN protein and plays a crucial role in the biogenesis of spliceosomes. We have identified three novel splicing variants of the SIP1 (SIP1-beta, -gamma and delta), in addition to the full-length SIP1-alpha. SIP1-alpha as found to be ubiquitously expressed at high levels in the various normal tissues examined. In contrast, SIP1-beta and -gamma were expressed at very low levels in these tissues. In muscle specimens from patients with spinal muscular atrophy (SMA) and amyotrophic lateral sclerosis (ALS), the expression of SIP1-alpha was dramatically decreased compared to that observed in the normal tissues. In addition, the expression of SIP1-beta was significantly increased in tissues derived from patients with either disease. These findings suggest that an aberrant alternative splicing event in SIP1 occurs tissues derived from patients with the motor neuron diseases, and contributes to the pathological process of SMA and ALS. PMID- 11943604 TI - The protective effect of calcium against genotoxicity of lead acetate administration on bone marrow and spermatocyte cells of mice in vivo. AB - The protective effect of calcium given orally by gavage with two doses (40 and 80mg/kg body weight) was evaluated against clastogenecity induced by lead acetate with two concentrations (200 and 400mg/kg diet) on bone marrow and spermatocyte cells of mice in vivo. The parameter screened was percentage of chromosomal aberrations with and without gaps and sperm abnormalities. Statistical analyses indicated the protection efficacy of calcium with the high dose rather than the other in both types of mouse cells. The observation from the laboratory tests, dealing that lead acetate can be considered as an environmental genotoxic material. We recommended that it must be administered of calcium (as calcium chloride) as a protective agent to reduce the genotoxic effect of lead in the somatic and germ cells. PMID- 11943605 TI - Cytogenetic studies on Nigella sativa seeds extract and thymoquinone on mouse cells infected with schistosomiasis using karyotyping. AB - The protective effect of Nigella sativa seed extract and its main constituents thymoquinone (TQ) was studied on mouse cells infected with schistosomiasis. Bone marrow cells in the in vivo experiments and spleen cells in the in vitro one were used to evaluate the potentially protective effect of these natural compounds on the induction of chromosomal aberrations. Karyotyping of the mice cells illustrated that the main abnormalities were gaps, fragments and deletions especially in chromosomes 2, 6 and some in chromosomes 13 and 14. Both N. sativa extract and TQ were considered as protective agents against the chromosomal aberrations induced as a result of schistosomiasis. PMID- 11943606 TI - Tea polyphenols inhibit the formation of mutagens during the cooking of meat. AB - Powerful mutagens are formed during the broiling or frying of meat. These mutagens cause specific cancers in animal models, and epidemiological studies suggest that they increase the risk of breast and colon cancer. It is important, therefore, to inhibit the formation of these mutagens. Application of tea polyphenols, polyphenon 60 from green tea, and polyphenon B from black tea, to both surfaces of ground beef before cooking inhibits the formation of the mutagens in a dose-related fashion. This procedure is simple and effective, and utilizes inexpensive tea, a product that deserves consideration for practical use. PMID- 11943607 TI - Induction of micronuclei by alkaloids extracted from Senecio brasiliensis and stored for 23 years. AB - In the present study, we report the results of an investigation on pyrrolizidine alkaloids extracted from Senecio brasiliensis (Sprengel) Less., which were stored for more than 23 years under variable conditions of temperature and humidity and exposed to light. Both the crude alkaloid (integerrimine+retrorsine+impurities) and pure integerrimine conserved the ability to induce acute toxicity in mice, leading to the death of the animals in less than 24h. The alkaloids also conserved the potential to induce significant increases in micronucleus frequencies in polychromatic erythrocytes of mouse bone marrow compared to the negative control. The administration of alkaloids to lymphocyte cultures blocked with cytochalasin-B showed no significant increase in micronucleus frequency in binucleated cells, probably due to the lack of a metabolic activation mechanism. However, an antimitotic effect was observed. PMID- 11943608 TI - Enhanced frequency of micronuclei in individuals exposed to arsenic through drinking water in West Bengal, India. AB - In West Bengal, India arsenic in ground water has been found to be above the maximum permissible limit in seven districts covering an area of 37,493km2. In the present study, evaluation of the micronuclei (MN) formation in oral mucosa cells, urothelial cells and peripheral blood lymphocytes was carried out in the symptomatic individuals exposed to arsenic through drinking water. Forty five individuals with cutaneous signs of arsenicism from four affected districts (368.11 microg/l of As in drinking water) were considered as the exposed group and 21 healthy individuals with no symptoms of arsenic poisoning and residing in two unaffected districts (5.49 microg/l of As) were considered as controls. The exposed and control groups had similar age distribution and socioeconomic status. Standardised questionnaires were utilised and medical examination was conducted to ascertain exposure history, sociodemographic characteristics, diet, health, medication, addiction and chief symptoms in the study participants. Arsenic exposure was confirmed by measuring the arsenic content in the drinking water, nails, hair and urine samples from the volunteers. Arsenic contents in the urine, nail and hair in the exposed group were 24.45 microg/l, 12.58 and 6.97 microg/g, respectively which were significantly high in comparison to corresponding control group values of 4.88 microg/l, 0.51 and 0.34 microg/g, respectively. Exposed individuals showed a statistically significant increase in the frequency of MN in oral mucosa, urothelial cells and lymphocytes (5.15, 5.74 and 6.39/1000 cells, respectively) when compared with the controls (0.77, 0.56 and 0.53/1000 cells, respectively). Thus, the above results indicate that the symptomatic individuals exposed to arsenic through drinking water in this region have significant cytogenetic damage. PMID- 11943609 TI - Analyses of bulky DNA adduct levels in human breast tissue and genetic polymorphisms of cytochromes P450 (CYPs), myeloperoxidase (MPO), quinone oxidoreductase (NQO1), and glutathione S-transferases (GSTs). AB - Environmental carcinogens are converted into DNA-reactive metabolites by phase I and phase II enzymes that are involved in the activation and detoxification of xenobiotics. Several of these enzymes display genetic polymorphisms that alter their activity leading to individual variation in DNA damage levels and thus cancer susceptibility. We investigated the relationship between DNA adduct levels and genetic polymorphisms in key enzymes of chemical carcinogenesis: CYP1A1, CYP1A2, GSTT1, GSTM1, GSTP1, NQO1 and MPO. Levels of DNA adducts were determined in human breast tissue using the 32P-postlabeling method. A significantly higher adduct level was observed for individuals with the A-463 variant in the MPO gene (P=0.008), providing the first observation of an association between a predicted reduced MPO gene transcription and a higher level of DNA adducts. Furthermore, levels of DNA adducts were about 45% higher in individuals with either GSTP1*B or GSTP1*C variants compared to those homozygous for the wild-type allele. When the MPO and GSTP1 were examined together, individuals with these combined variant genotypes had significantly higher adduct levels than all other genotype combinations (P=0.003). PMID- 11943610 TI - A combination of the micronucleus assay and a FISH technique for evaluation of the genotoxicity of 1,2,4-benzenetriol. AB - The cytokinesis-block micronucleus (CBMN) assay has emerged as one of the preferred methods for assessing chromosome damage. Micronuclei (MN) are small, extranuclear bodies that are formed in mitosis from acentric chromosomal fragments or chromosomes that are not included in each daughter nucleus. Thus, MN contain either chromosomal fragments or whole chromosomes. The CBMN assay, together with a fluorescence in situ hybridization (FISH) technique using specific centromeric probes for chromosomes 7 and 8, were employed in mitogen stimulated human lymphocytes pretreated with the benzene metabolite, 1,2,4 benzenetriol (BT). Treatment of human lymphocytes resulted in the induction of MN in a dose-dependent manner. The frequency of MN in control lymphocytes was 4.5 per 1000 binucleated (BN) cells and this increased to 9.5, 14, 28 and 40 per 1000 BN cells at 10, 25, 50 and 100 microM BT, respectively. The frequency of aneuploidy 7 and 8 in BN cells also increased at each concentration. Aneuploidy 8 was more frequent than aneuploidy 7, suggesting that chromosome 8 is more sensitive to aneuploidy induction by BT. The frequency of MN containing centromere positive signals for chromosomes 7 and 8 increased with the concentration of BT. The frequency of MN with centromere positive signals was higher for chromosome 8 than for chromosome 7, also suggesting a greater sensitivity of chromosome 8 to this agent. These results suggest that combined application of the CBMN assay with a FISH technique, using chromosome-specific centromeric probes, would allow the detection of aneuploidy in human lymphocytes and identify the mechanistic origin of MN induced by a clastogen or aneugen. PMID- 11943611 TI - Should gaps be included in chromosomal aberration analysis? Evidence based on the comet assay. AB - This study evaluated DNA damage in human lymphocytes due to occupational exposure to low levels of ionizing radiation using two assays: the comet assay and chromosomal aberration (CA) analysis including and excluding gaps. The results obtained reveal a higher correlation between both methods when chromatid and chromosome gaps were included in the correlation analysis (r=0.78 versus r=0.50). This increased correlation support the hypothesis that the gaps constitute a type of chromosome aberration, and suggest that these events should be scored in this type of analysis. PMID- 11943612 TI - Biomonitoring of primary aluminium industry workers: detection of micronuclei and repairable DNA lesions by alkaline SCGE. AB - The genetic effects of occupational exposure to low polycyclic aromatic hydrocarbon (PAH) concentrations were investigated in primary aluminium industry workers. The study subjects were employed in a plant that uses pre-baked anode cells, and has relatively low PAH contamination. Forty-two male workers belonging to different job categories (anode fabrication, baking, rodding, electrolysis, maintenance), together with 16 male local residents with no occupational exposure to PAHs were selected for the analysis of micronuclei and DNA lesions in peripheral lymphocytes. The incidence of micronuclei determined in 1000 cytokinesis-blocked cells in each subject was not significantly different between workers and controls (8.5+/-5.4 per thousand versus 9.7+/-4.9 per thousand, respectively), nor between smokers and non-smokers (8.3+/-5.8 per thousand versus 9.2+/-5.1 per thousand), but was significantly (P<0.05) related to the subjects' age. Also the analysis of DNA damage in unstimulated and mitogen-stimulated lymphocytes by single cell gel electrophoresis (SCGE) did not show significant differences between the studied groups (average tail moment values were 0.53+/ 0.53 and 0.49+/-0.45 microm in exposed subjects and controls, respectively). However, when lymphocytes were cultured in the presence of cytosine arabinoside (Ara-C, 1 microg/ml for 16h) the SCGE analysis revealed a significant (P=0.018) difference in tail moment values between aluminium workers and controls (1.73+/ 1.05 microm versus 0.93+/-0.88 microm, respectively). This difference may highlight an excess of relatively stable DNA lesions, that do not affect strand integrity, and are expressed as intermediates of excision repair in stimulated cells, when gap refilling is inhibited by cytosine arabinoside (Ara-C). PMID- 11943614 TI - The influence of the solvents DMSO and ethanol on the genotoxicity of alpha,beta unsaturated aldehydes in the SOS chromotest. AB - alpha,beta-Unsaturated aldehydes are ubiquitous environmental pollutants, important industrial chemicals, have mani-fold biological functions in plants and insects and are natural products in food. They are endogenously formed in animals and humans during lipid peroxidation and arachidonic acid oxidation and are genotoxic, mutagenic and carcinogenic. Crotonaldehyde and 2-hexenal in food may contribute to general carcinogenicity in humans. The high bacterial toxicity of these compounds leads to problems in genotoxicity testing in bacterial systems. Recently, we have shown that using ethanol as solvent instead of dimethylsulfoxide (DMSO) results in an increase in the induction factors and the SOS-inducing potency of alpha,beta-unsaturated ketones in the SOS chromotest. Here, we demonstrate that utilization of ethanol as solvent also improves the testing of alpha,beta-unsaturated aldehydes. Five aldehydes out of nine tested were clearly positive in the SOS chromotest according to the criteria of Quillardet, i.e. acrolein, crotonaldehyde, 2,4-hexadienal, 2-methylacrolein and 2 ethylacrolein, three further, 2-hexenal, 2-heptenal and 2-propylacrolein showed a dose dependent increase of the induction factors which was however lower than 1.5 times that of the background. Only 2-butylacrolein did not lead to an increase in the induction factors. With DMSO as solvent only the three aldehydes acrolein, crotonaldehyde and 2,4-hexadienal showed an increase in the induction factor, which was however lower than 1.5 that of the background. Utilization of ethanol allows to establish structure genotoxicity relationships for alpha,beta unsaturated aldehydes in the SOS chromotest. Genotoxicity decreases with increasing degree of substitution. The decreasing genotoxicities can be explained (a) by increasing bacterial toxicity due to increasing lipophilicities of the higher substituted aldehydes and (b) by decreasing reactivity due to steric hindrance by the alkyl substituents. PMID- 11943613 TI - Changes in mutagenicity of herbicide chlornitrofen during biodegradation. AB - We used the Ames assay to investigate changes in the mutagenicity of chlornitrofen during its aerobic biodegradation. Although a mixed culture of bacteria obtained from river water degraded chlornitrofen and reduced its concentration from 39 to 6 microg/l in 21 days, the indirect mutagenicity of the solution to Salmonella strains TA98, YG1021, and YG1026 increased gradually. This finding suggests that mutagenic metabolites were produced during the aerobic biodegradation. The increase in the mutagenicity was, however, much smaller under aerobic than under anaerobic conditions. The differing sensitivities of our test strains to the functional groups on the mutagens showed that the mutagenic metabolites were indirect frameshift-type mutagens that might have neither nitro nor amino groups. PMID- 11943615 TI - DNA adducts as indicators of genotoxic exposure in indigenous and transplanted mussels, Mytilus edulis L. from Icelandic coastal sites. AB - Indigenous mussels, Mytilus edulis, were collected at sites with supposed different amounts of pollution; Reykjavik harbour, Keflavik harbour, Grafarvogur and Hvalfjordur (reference), along the south-western coast of Iceland in March 2000. Mussels from Hvalfjordur and Reykjavik harbour were also collected in August the same year. Additionally, mussels were transplanted from the reference site to Reykjavik harbour for 6 weeks during both winter and summer for comparison. DNA adducts were analysed by 32P-post-labelling in gills and digestive gland. Highest adduct levels were found in gill tissue from indigenous mussels collected in Reykjavik harbour. Adduct levels in both tissues from mussels collected at the reference site were below or very close to the detection limit during winter, but seemed to increase a little during summer. Mussels from sites with supposed intermediate pollution had intermediate levels of DNA adducts in gills but did not differ from Reykjavik harbour in digestive gland. No increase in adduct levels was observed in mussels transplanted from the reference site to Reykjavik harbour, except for a slight increase in digestive gland during winter. This study shows that 32P-post-labelling analysis of DNA adducts is sensitive enough to be used on indigenous mussels from relatively pristine areas and that adduct levels are increased in harbours/urban sites. However, transplantation of mussels from a clean site to the harbour for 6 weeks did not result in increased adduct levels in gills, the tissue with the highest adduct levels. The results also indicate that seasonal variation in adduct levels may occur. PMID- 11943616 TI - Hemoglobin adducts and micronucleus frequencies in mouse and rat after acrylamide or N-methylolacrylamide treatment. AB - The reactive industrial chemicals acrylamide (AA) and N-methylolacrylamide (MAA) are neurotoxic and carcinogenic in animals, MAA showing a lower potency than AA. The causative agent in AA-induced carcinogenesis is assumed to be the epoxy metabolite, glycidamide (GA), which in contrast to AA gives rise to stable adducts to DNA. The causative agent in MAA induced carcinogenesis is so far not studied. The two AAs were studied in mice and rats using analysis of hemoglobin (Hb) adducts as a measure of in vivo doses and the in vivo micronucleus (MN) assay as an end-point for chromosome damage. Male CBA mice were treated by intraperitoneal (i.p.) injection of three different doses and male Sprague-Dawley rats with one dose of each AA. Identical adducts were monitored from the two AAs [N-(2-carbamoylethyl)valine] and the respective epoxide metabolites [N-(2 carbamoyl-2-hydroxyethyl)valine]. Per unit of administered amount, AA gives rise to higher (three to six times) Hb adduct levels than MAA in mice and rats. Mice exhibit, compared with rats, higher in vivo doses of the epoxy metabolites, indicating that AAs were more efficiently metabolized in the mice. In mouse the two AAs induced dose-dependent increases in both Hb adduct level and MN frequency in peripheral erythrocytes. Per unit of administered dose MAA showed only half the potency for inducing micronuclei compared with AA, although the MN frequency per unit of in vivo dose of measured epoxy metabolite was three times higher for MAA than for AA. No increase in MN frequency was observed in rat bone marrow erythrocytes, after treatment with either AA. This is compatible with a lower sensitivity of the rat than of the mouse to the carcinogenic action of these compounds. PMID- 11943617 TI - High yield of endoreduplication induced by ICRF-193: a topoisomerase II catalytic inhibitor. AB - An uncommonly high yield of spontaneous endoreduplication is a feature of the CHO mutant EM9, besides its defective repair of single, as well as double-DNA strand breaks and its extraordinarily elevated yield of sister chromatid exchanges (SCEs) after bromodeoxyuridine (BrdU) incorporation into DNA. Since the nuclear enzyme topoisomerase II (topo II) has been reported to be responsible for the segregation of daughter chromosomes during mitosis, in the present investigation we have made use of the bisdioxopiperazine ICRF-193, a topo II catalytic inhibitor that interferes with the normal turnover of the enzyme. In order to see whether both EM9 cells and its parental cell line AA8, which show differences in the spontaneous frequency of endoreduplicated cells are or not equally sensitive to the topo II catalytic inhibitor, both cell lines have been treated with a range of doses of the bisdioxopiperazine. Our results show that both cell lines respond to the treatment entering in an endoreduplication cycle, but the EM9 cells are extremely sensitive to the inhibition of topo II. PMID- 11943618 TI - Detection of radiation and cyclophosphamide-induced mutations in individual mouse sperm at a human expanded trinucleotide repeat locus transgene. AB - A method to measure the germline mutations induced by cancer treatment in humans is needed. To establish such a method we used a transgenic mouse model consisting of a human DNA repeat locus that has a high spontaneous mutation frequency as a biomarker. Alterations in repeat number were measured in individual sperm from mice hemizygous for an expanded (CTG)(162) human myotonic dystrophy type 1 (DM1) microsatellite repeat using single genome-equivalent (g.e.) PCR and detection by a DNA fragment analyzer. Mutation frequencies were measured in DNA from sperm from controls and sperm derived from stem spermatogonia, differentiating spermatogonia, and spermatocytes exposed to radiation and from spermatocytes of mice treated with cyclophosphamide. There was no increase above control levels in mutations, scored as >1 repeat changes, in any of the treated groups. However, moderately large deletion mutants (between 9 and 20 repeat changes) were observed at frequencies of 2.2% when spermatocytes were treated with cyclophosphamide and, 1.8 and 2.5% when spermatocytes and stem cells, respectively, were treated with radiation, which were significantly higher than the frequency of 0.3% in controls. Thus, radiation and cyclophosphamide induced deletions in the expanded DM1 trinucleotide repeat. PCR artifacts were characterized in sperm DNA from controls and from mice treated with radiation; all artifacts involved losses of more than 20 DM1 repeats, and surprisingly the artifact frequency was higher in treated sperm than in control sperm. The radiation-induced increase in the frequency of PCR artifacts might reflect alterations in sperm DNA that destabilize the genome not only during PCR amplification but also during early embryonic development. PMID- 11943619 TI - Assessment of genotoxic effects of chloropyriphos and acephate by the comet assay in mice leucocytes. AB - Two organophosphorus (OP) pesticides (chloropyriphos and acephate) and cyclophosphamide (CP) (positive control) were tested for their ability to induce in vivo genotoxic effect in leucocytes of Swiss albino mice using the single cell gel electrophoresis assay or comet assay. The mice were administered orally with doses ranging from 0.28 to 8.96 mg/kg body weight (b. wt.) of chloropyriphos and 12.25 to 392.00 mg/kg b.wt. of acephate. The assay was performed on whole blood at 24, 48, 72 and 96 h. A significant increase in mean comet tail length indicating DNA damage was observed at 24h post-treatment (P<0.05) with both pesticides in comparison to control. The damage was dose related. The mean comet tail length revealed a clear dose dependent increase. From 48 h post-treatment, a gradual decrease in mean tail length was noted. By 96 h of post-treatment the mean comet tail length reached control levels indicating repair of the damaged DNA. From the study it can be concluded that the comet assay is a sensitive assay for the detection of genotoxicity caused by pesticides. PMID- 11943620 TI - Evaluation of DNA damage in flight personnel by Comet assay. AB - There have been some suggestions that air-crew are at a higher-than-normal risk of developing cancer, since they are exposed to potential genotoxic factors. These include cosmic radiations, airborne pollutants such as the combustion products of jet propulsion, ozone, and electromagnetic fields. We used the Comet assay to investigate DNA damage in flight personnel with the aim of assessing potential health hazards in this occupational category. We studied 40 civil air crew members who had been flying long-haul routes for at least 5 years, and compared them with a homogeneous control group of 40 healthy male ground staff. The Comet assay, or single-cell gel electrophoresis (SCGE), detects DNA single- and double-strand breaks (DSBs) and alkali-labile lesions in individual cells, and is a powerful and sensitive technique for detecting genetic damage induced by different genotoxic agents. Taking into consideration occupational risk and possible confounding factors, this assay showed a small increase, that did not reach statistical significance, of DNA damage in long-haul crew members compared to controls, indicating a lack of evident genotoxic effects. An association, although again not statistically significant, was found between reduced DNA damage and use of protective drugs (antioxidants). PMID- 11943632 TI - [Nutrition and development in the era of globalization: new challenges for public health]. PMID- 11943633 TI - [Iodine deficiency: current situation and future prospects]. AB - Iodine deficiency disorders (IDD) is a major public health problem worldwide. WHO estimates that 740 million people are currently affected by goitre. The consequences of iodine deficiency on health are the results of hypothyroidism and the main one is impaired development of foetal brain. IDD is the first cause of preventable brain damage in children. The recommended strategy to correct IDD rests upon salt iodisation. Over the last 20 years, the international community mobilised to eliminate IDD under the leadership of WHO, Unicef and ICCIDD. It resulted in remarkable progress in IDD control, especially in Africa and in South East Asia where the endemic is the most severe. It is estimated that 68% of the populations of affected countries have currently access to iodised salt. However, out of the 130 affected countries, about 30 have no programme. Besides, salt quality control and monitoring of population iodine status are still weak in many countries, thus exposing the population to an excessive iodine intake and subsequently to the risk of iodine-induced hyperthyroidism. In addition, IDD is re-emerging in some countries, especially in Eastern Europe after it had disappeared. In order to reach the goal of IDD elimination, it is important to insist on the sustainability of salt iodisation programmes, which implies an increased commitment of both health authorities and representatives of the salt industry. PMID- 11943634 TI - [Assessment of iodine content of dietary salt in Abidjan (Cote d'Ivoire)]. AB - Cote d'Ivoire is one of the countries engaged in the strategy of universal salt iodization set up to prevent and control iodine deficiency disorders. However, no systematic monitoring of iodine content of salt has been performed up to now. Therefore, a survey was conducted on a random sample of 400 households in the Marcory district of Abidjan in order to study consumers' behaviour regarding the purchase and storage of salt, and to determine the iodine content of dietary salt. The proportioning of iodine was carried out by the titrimetric method. The salt consumed in the homes was bought at the market, the shop, the supermarket or was delivered at residence by a hawker in the proportions of 82.8%, 10.3%, 6.3%, and 0.8%, respectively. At the warehouse, the salt purchased was packaged into bags of 25 kg for 0.5% of homes. 1% of homes bought salt in tins. 9.5% of homes bought their salt in sachets. Most homes (89%) replenished their stock in bulk from the market. Laboratory analyses showed that all salt samples were iodised. The average content of iodine in salt was 52.74 ppm with a standard deviation of 32.56. We observed that in 23.3% of households the iodine content of salt was weak (< 30 ppm). Contents higher than the upper limit of normality (30-50 ppm) were found in 44.8% of cases. The level of iodine was adequate for 32% of households. The major risk to which the population is exposed is the outbreak of iodine-induced hyperthyroidism (IIH) despite the fact that the Abidjan area is not naturally deprived of iodine. The risk is even more significant in the goitre endemic zone of the western and northern areas of the country, because these regions receive the same salt as Abidjan. From this arises the need for setting up an effective system of controlling the iodine content in dietary salt to continuously adjust it, taking into consideration the minimum needs of the population. PMID- 11943635 TI - [Control of iron deficiency in developing countries]. AB - Iron deficiency is the most prevalent nutritional disorder worldwide, especially in developing countries. It occurs when iron absorption cannot compensate iron requirements and losses. Requirements are especially high in pregnant women, infants, young children and adolescents who run a higher risk of being iron deficient. In developing countries, the main cause of iron deficiency is the low iron bioavailability of the diet. The consequences of iron deficiency are many and serious, affecting not only individuals' health but also the development of societies and countries. The prevention and the control of iron deficiency and anemia in all groups of a population with different iron requirements imply to coordinate different interventions. Iron fortification of staple foods or condiments directed to the whole population is a sustainable and low cost effective approach. However, at some periods of life, especially during pregnancy and in children from the age of 6 months, iron requirements are high. For pregnant women, the current approach favours the daily iron-folate supplementation during pregnancy but the results in terms of public health are disappointing. The preventive weekly iron-folate supplementation of women during their reproductive life, whose efficacy is recognized, offers a promising alternative; its impact in terms of public health is under current evaluation. For infants and young children, iron fortification of complementary food is effective but this food is generally imported and economically inaccessible to populations with limited resources. The production, by small private units from local products, of complementary foods of low viscosity, good nutritional quality, fortified with vitamins and minerals, and of low cost is at hand in several countries. When complementary foods are not available, the preventive iron supplementation from 6 to 18 months of age has to be advised. This approach should be strengthened by the advantages of the weekly approach. These interventions are more effective when they integrate other approaches like the improvement of the nutritional practices, infection control and the promotion of breast-feeding and when coupled with programs aiming to control other micronutrient deficiencies. The success of most interventions requires the active participation of the individuals. Information and education of the populations, especially through social mobilization campaigns, are essential because iron deficiency induces few visible symptoms, not easily recognizable by individuals. The implementation of national nutrition plans including the control of iron deficiency as one of the priorities and the participation of the public health and education sectors, food industries, the community and the media should contribute to the success of the interventions and to the control of iron deficiency. PMID- 11943637 TI - [Changes in vitamin A intake following the social marketing of red palm oil among children and women in Burkina Faso]. AB - This paper focuses on changes in vitamin A (VA) intakes as part of the evaluation of a pilot project on social marketing of red palm oil (RPO) as a VA supplement for mothers and children in central-north Burkina Faso. The objectives of the 30 month project are to demonstrate the feasibility and effectiveness of introducing RPO in non-consuming areas. RPO is collected from women in the South-West region and it is sold in project sites by village volunteers. RPO is promoted by community workers trained in persuasive communication and social marketing. The target population is free to buy and consume RPO. Evaluation design includes data collected at onset, then 12 and 24 months later, from the same sample of 210 mothers and their children randomly selected in seven project sites. Children were 1 to 3 years old at onset. Blood samples were collected at baseline from mothers and children for serum retinol determination by HPLC. VA intakes are estimated by a semi-quantitative food frequency questionnaire, using the conventional beta-carotene to retinol conversion factors and the newly revised lower factors. VA deficiency is a major public health problem in the area: 64% of mothers and 85% of children had serum retinol concentrations < 0,70 mumol/l at baseline. VA came mainly from plant foods, particularly fruits and dark green vegetables which provided more than 90% of the dietary VA at onset of the project. Mean vitamin A intakes are low. We found 138 106 mug ER for the children and 302 +/- 235 microg ER for the mothers with conventional factors and 64 +/- 58 microg ER and 133 +/- 162 microg ER, respectively, with the revised factors. One year later, one third of respondents had consumed RPO in the previous week, and it supplied around 56% of the VA intake of children and 67% of mothers (36% and 46% respectively for the whole group). VA intakes were significantly increased at 510 +/- 493 microg ER and 801 +/- 913 microg ER for the children and their mothers respectively (347 +/- 443 microg ER and 568 +/- 803 microg ER respectively, with the revised factors). Analyzing serum retinol and dietary data collected at baseline, it was found that VA intakes < 62,5% of safe level of intake were highly sensitive to low serum retinol (< 0,70 micromol/l) and using revised conversion factors to assess total VA intake slightly enhanced sensitivity. The proportion of mothers and children at risk of inadequate VA intake changed from nearly 100% at baseline to 60% one year later. The results show that promoting RPO (and other VA rich foods) was effective in improving VA intakes. This improvement will hopefully be sustained and even further enhanced during the remaining 12 months of the project, after which repeated measurement of serum retinol and VA intakes will allow the actual impact of the project to be truly assessed. PMID- 11943636 TI - [Efficacy of daily and weekly iron supplementation for the control of iron deficiency anaemia in infants in rural Vietnam]. AB - In Vietnam the high prevalence of iron deficiency anaemia in infants and young children speaks for implementing early interventions. This study aimed to evaluate the efficacy of the daily iron supplementation in infants given by their mothers and of the weekly iron supplementation. Two hundred and seventy infants aged 5 to 12 months, were divided into four groups. Group 1 received a placebo daily and group 2 a daily dose of 15 mg iron (2.0 +/- 0.3 mg iron/day/kg body weight) which was given by their mothers for three months. Group 3 received a daily dose of 15 mg iron and group 4 a weekly dose of 15 mg iron given during 6 months by health auxiliaries. The hemoglobin concentration (Hb) was measured before the intervention and after 3 months of supplementation in all groups, and again after 6 months of supplementation in groups 3 and 4. After 3 months of iron supplementation, the hemoglobin concentration increased significantly by 21.6 +/- 12.3 g/l and the prevalence of anaemia decreased from 81.3% to less than 9% in group 2. The weekly iron supplementation was significantly less effective than the daily supplementation: after 3 months, Hb increased by 15.4 +/- 13.3 g/l in group 3 and by 11.2 +/- 10.2 g/l in group 4 (p = 0.04) and the prevalence of anaemia was 17.9% and 41.5% in groups 3 and 4, respectively. After 6 months of supplementation, changes in Hb were not significantly different between group 3 (22.0 +/- 12.1 g/l) and group 4 (20.0 +/- 10.0 g/l, p = 0.30). However, the final hemoglobin concentration was significantly lower in group 4 (120.5 +/- 7.2 g/l) than in group 3 (123.6 +/- 7.8, p = 0.02). Moreover, nearly 8% of the children were still anemic in group 4 versus 0% in group 3. Since the early developmental period, when the brain and other specific organs are especially sensitive to iron deficiency, is critical, 3-month daily iron supplementation of infants from the age of 6 months has to be recommended, followed by a weekly iron supplementation until the age of 15 months. The community approach, where mothers informed on the importance of iron deficiency anaemia and on the consequences for the health of their infants gave the iron supplements, was shown to be effective. However, its sustainability would depend on the availability of low-cost iron supplements affordable by populations with limited economic resources. Other interventions, such as iron supplementation of women during fertile age, especially during pregnancy and lactation periods, and the use of complementary food to breast milk, fortified with micronutrients, should be associated. PMID- 11943638 TI - [Nutritional transition and non-communicable diet-related chronic diseases in developing countries]. AB - It is increasingly recognized that developing countries are undergoing an epidemiologic transition similar to that which occurred in industrialized countries in previous centuries. While infectious diseases are still the main cause of morbidity and mortality, there is a marked increase in chronic non communicable diseases, particularly in the most advanced developing countries, and these diseases are expected to take the lead in a decade or two. Most of these diseases, above all coronary heart diseases, stroke and diabetes, are related to diet and lifestyles, for example tobacco and alcohol consumption. As a matter of fact, these societies are also facing a growing epidemic of overweight and obesity, due to the frequent energetic imbalance between energy-dense food consumption and reduced daily physical expenditure. This health transition, favoured by demographic changes towards aging populations, is occurring at an increased pace in urban societies widely exposed to the modernization of lifestyle, sedentary occupation, and to lipid- and sugar-rich food, often poor in fibre and micronutrients. Increased world access to cheaper vegetable oil is thought to have triggered off this accelerated and generalized trend, though animal food, rich in saturated fat, and imported or locally-made industrialized food also play a role. While increased national and household incomes facilitate the initial change, as the transition advances poor people progressively become the main victims, as has been observed in the more advanced developing countries. Metabolic imprinting due to intra-uterine and infant malnutrition, which are still common in these societies, is also thought to play a significant role in the increase in the expression of insulin resistance, obesity and chronic diseases when these children are exposed to abundant food and modern lifestyle, later in life. Treatment and secondary prevention of nutrition-related chronic diseases and associated disabilities have an ever rising cost in industrialized countries, which is far beyond the means of the still fragile economies of developing countries. This double burden of infectious diseases and undernutrition that still exist, and of non-communicable diseases and overnutrition represents a threat to the frequently unprepared health care services in developing countries. There is a clear need to focus health policies on the prevention of chronic diseases through primary health care services, the use of mass media for communication and education about healthy nutrition and lifestyle, and the adaptation of public policies. Nutritionists must also adapt to this changing nutritional situation which may result in apparently contradictory nutritional status findings within societies if not even within households. PMID- 11943639 TI - [Foetal programming of nutrition-related chronic diseases]. AB - Intrauterine growth retardation, which reflects in large part maternal malnutrition in poorer communities, contributes to chronic disease risk through foetal programming, according to the early origins hypothesis of Barker. Foetal programming implies that during critical periods of prenatal growth, permanent changes in metabolism or structures result from adverse intrauterine conditions. Observational studies first showed an association between lower birth weights and higher rates of coronary disease in the 80s, in England and Scandinavia. The link between low birth weights, or other indicators of small birth size, and cardiovascular disease was later confirmed in many epidemiological studies, including in the USA and in India. Similarly, a reverse relationship of birth weight and systolic blood pressure was shown in men and women, in developed as well as developing countries, and in all age groups, although it was less consistent in adolescents. Insulin resistance and type-2 diabetes have also been found to be independently related to small size at birth in several studies around the world. Insulin resistance associated with small size at birth was frequently shown to be present at a young age. The association of small birth size with chronic disease tends to increase with catch-up growth and obesity, and usually persists after adjusting for confounding factors such as age, family history, and socio-economic status. Several, but not all, twin studies lend support to the hypothesis. There is a tendency for lighter members of twin pairs to have a higher blood pressure, and more diabetes. Observations in people exposed to the Dutch famine while in utero also tend to corroborate the hypothesis. Those who were exposed early in their intrauterine life did not have lower birth weights, but they were prone to becoming obese later on. In contrast, those exposed towards the end of gestation had lower birth weights, and showed a higher rate of impaired glucose tolerance, while having a lower risk of obesity. Dietary manipulations in animal models provide further support and mechanistic explanations, in particular protein deficiency in pregnant rats, which elevates blood pressure, impairs glucose tolerance, and increases the likelihood of obesity in the progeny. Although there are still controversial areas, there is at present sufficient scientific evidence for foetal programming to be regarded as an additional risk factor for chronic disease, in interaction with genetic and lifestyle risk factors. The fact that intrauterine growth retardation may predispose to nutrition-related chronic disease has serious implications for developing countries, particularly those undergoing rapid nutritional transition, as it may further increase the rates of obesity, cardiovascular disease and diabetes when diets and lifestyles are in themselves "atherogenic". The challenge is for programmes to simultaneously combat apparently opposite nutrition problems, malnutrition and "over-nutrition". Improving the nutrition of women is even more imperative when considering that it may contribute to preventing chronic diseases in the next generation, in addition to enhancing health and survival of mothers and children. PMID- 11943640 TI - [Breastfeeding in Africa: will positive trends be challenged by the AIDS epidemic?]. AB - In Africa, more than 95% of infants are currently breastfed, but feeding practices are often inadequate: feeding water, and other liquids, to breastfed infants is a widespread practice. Consequently, the rate of exclusive breast feeding is low, particularly in West Africa. The rate of bottle-feeding is high in some countries (exceeding 30% in Tunisia, Nigeria, Namibia and Sudan). Nevertheless prolonged breastfeeding is common, and the median duration of breastfeeding ranges between 16 and 28 months. Urbanization and mothers' education are the major factors that tend to shorten breastfeeding. Nevertheless recent trends show an increase in early initiation and in duration of breastfeeding as a result of promotion efforts deployed by WHO and Unicef, local governments, and non-governmental organizations. The importance of breastmilk as a food resource of African countries is generally not recognized. In 31 countries where data on prevalence of breastfeeding are available, consumption of breastmilk by children under three years is estimated at 3.5 million tons per year. The AIDS epidemic could threaten breastfeeding because the virus can be transmitted through breastmilk, as demonstrated by numerous studies. A study suggests that feeding breastmilk and other liquids to infants could be the feeding mode associated with the highest rate of transmission. To prevent mother to-child transmission of HIV, WHO recommends replacement feeding if it is feasible and safe. Otherwise, mothers are encouraged to practice exclusive breastfeeding for the first months of life followed by early and rapid weaning. The feasibility of replacement feeding with breastmilk substitutes, however, is very uncertain. In a study where free substitutes were given to HIV-positive mothers, the mortality of the formula-fed infants was the same as that of the breastfed infants. HIV-positive mothers may find it difficult to cope with the constraints of replacement feeding, in terms of cost, workload and time, and with the additional health care needs of non-breastfed infants. Exclusive breastfeeding for a few months could carry a lower risk of death than replacement feeding. But success in promoting exclusive breastfeeding has been limited in Africa, and new promotion methods are needed. Infants of all mothers, whether HIV positive or not, will benefit from improving the rate of exclusive breastfeeding. The major problem is to ensure that early and rapid weaning, between 4 and 6 months, does not have a negative impact on the child's health. Early weaning is known to increase susceptibility to infections and can cause malnutrition. The feasibility and safety of this recommendation will have to be monitored carefully. A strong determination of African governments to promote exclusive breastfeeding among all mothers and to protect prolonged breastfeeding among non infected mothers will limit the mother-to-child transmission of HIV while preserving the benefits of breastfeeding. PMID- 11943641 TI - [What if the HIV/AIDS pandemic were an opportunity to improve the relevance of breastfeeding promotion programs in Africa?]. AB - In the 1990s, many programs and actions were set up for promoting breastfeeding in Africa, more or less successfully in different countries. The main achievements of these programs were the training of health professionals and the apparent ending of the distribution of formula in Health services. The impact of these programs on breastfeeding practices in countries with prevalent prolonged mixed feeding is less obvious, as many programs did not emphasize "best practices". Health messages delivered on this topic have been poor, because they were often conceived at the international level rather than adapted to African contexts, and because the consensus about the promotion of breastfeeding is so strong that the programs have rarely undergone a critical evaluation. The HIV/AIDS pandemic could be an opportunity to rethink these programs. "Baby friendly" Health services are now considered as the most knowledgeable to deal with breastfeeding in the context of HIV, through the reinforcement of the promotion of "best feeding practices" and through the follow-up of formula feeding for some HIV-positive mothers. To prevent HIV transmission, health messages will have to promote some practices that are useful for HIV-negative, as well as HIV-positive, mothers, such as exclusive breastfeeding, the prevention, early diagnosis and treatment of abscesses and mastitis, and the management of weaning - all strategies that were undervalued until now. For children of HIV positive mothers and for orphans, Health services will have to set up a medical follow-up of artificial feeding. These new goals mean that breastfeeding promotion programs will have to develop complementary strategies with an emphasis on care, coordinated with other vertical programs such as AIDS and malnutrition programs. PMID- 11943642 TI - [Socio-anthropological contributions to public nutrition: why, how and under which conditions?]. AB - Socio-anthropological research is being increasingly recognised as important in public nutrition research. It helps to understand practices and behaviours the significance of which can partially escape the nutritionist. It also permits to increase knowledge about food systems and their underlying social dynamics and representations and therefore contributes to the understanding of often neglected aspects of nutritional problems. In this paper the authors highlight how their discipline and its main research techniques can be efficiently involved in public nutrition research, without however neglecting their own specific methodological requirements. The first objective of the paper is to draw methodological lessons from experiences. A second objective is to familiarise researchers, who are not specialised in social sciences, with the main techniques used in the discipline. After a brief presentation of the underlying principles and objectives of socio anthropological research, the main data collection techniques used by socio anthropologists (observation, interview, focus groups) are described and their use is illustrated by concrete research examples from Congo, Bolivia, Nepal and Tunisia. A special emphasis is put on the requirements of qualitative data analysis. The discussion focuses on the choice of techniques, the enhancement of quality in qualitative research and on the main constraints and difficulties encountered in involving socio-anthropology in public nutrition. The limits, constraints and conditions of this implication and contribution are discussed. For the authors the issue of involving socio-anthropology in public nutrition should not be viewed in terms of instrumentalisation of one discipline by the other. The main avenue to reach a fruitful and efficient collaboration is respect and recognition of the methodological approaches (and underlying epistemological assumptions) of socio-anthropology (qualification of researchers, quality criteria, etc.). These requirements are necessary for socio-anthropology to produce qualitatively sound data and analysis. More important, the set up of common conceptual frameworks on the basis of an interdisciplinary dialogue has to be established from conception of the research up to its conclusion. PMID- 11943643 TI - ["Care" and public nutrition]. AB - In 1990, the Unicef conceptual framework for nutrition recognised the role of care, along with household food security and health services and environment, as one of the three underlying factors of child survival, growth, and development. This model has been adopted at a policy level at the International Conference on Nutrition (Rome, 1992) and over the past ten years the concept of care has been refined through literature reviews, consultative meetings and empirical works. "Care is the provision in the household and the community of time, attention, and support to meet the physical, mental, and social needs of the growing child and other household members". Basically, care refers to the actions of caregivers (mainly, but not only mothers) that translate food and health resources into positive outcomes for the child's nutrition. Even under circumstances of poverty, enhanced caregiving can optimise the use of resources to promote good nutrition. Care practices have been grouped into six categories: care for women, breastfeeding and child feeding practices, psychosocial care, food preparation, hygiene practices, household health practices. They cover a wide range of behaviours, are often culturally specific and are daily, repetitive, and time consuming activities. It must be underlined that the way care practices are performed (i.e., quality of care) is as important as the practices themselves. It has also been emphasised that children play a significant role in determining the quality of care that they receive, through an interactive process: an active child elicits more care from the caregiver, who is in turn more responsive. Care resources at household level have been described according to three categories: human (knowledge, beliefs, education, physical and mental health of the caregiver), economic (control on income, workload and time), and organisational (alternate caregivers, community support). But the availability of care also depends on support at the national or international level. As the mother is the primary caregiver, most of the obstacles to care are the constraints to the mothers, the most common characteristic of which being the low status of women in many societies. More studies are required to better understand the causal relationship between care and nutrition. Methods to measure the qualitative aspects of care and indicators that capture the complexity of care must be developed and cross-culturally tested. These will also be useful to design and monitor more effective interventions incorporating care. These programmes should first identify and support the good traditional care practices rather than simply ask for change; the activities proposed should not break the balance between the time women spend on care and the time they spend on work. Therefore one must be sure that enough resources are available. Finally, to achieve sustainable changes a participatory and comprehensive approach is definitely needed. PMID- 11943644 TI - [Management of malnutrition in preschool children: the role of primary health care services]. AB - Although the prevalence of malnutrition in developing countries is decreasing, it is still a major problem for many children under five. As socio-economic conditions are the main determinants, a final solution for this problem can only be envisaged in the long run. Still, short-term strategies need to be defined in order to relieve the sufferings of individual children and their families. Understanding the problem and consequently formulating intervention programs at the local level remains a complex and difficult issue. The first reason being that the process of malnutrition expresses itself in different forms and with variable consequences. A second reason making malnutrition a complex problem is that the primary causes -- the interaction between insufficient food supply and the frequent recurrence of infectious diseases -- are determined by a multitude of factors of different natures. This complexity -- of its expressions, effects, and causality -- makes it difficult to get a global vision and understanding of the problem, which clearly impedes the definition of rational and integrated intervention strategies. Nevertheless, a better understanding of the pathophysiology of malnutrition and of the factors that influence the growth process in preschool age, will help to better direct actions. To this effect, a conceptual model will be built, based on recent insight in the process of malnutrition within this age group. From this model, two lines of action for increasing the chances of preschool children to express their initial growth potential, become apparent. A first series of activities could tackle the process that, via wasting and recurrence of infections, leads to an increased mortality risk. As timely intervention reduces the risk of depletion of energy reserves, these activities would also have an indirect impact on physical development. Elaborating strategies for secondary prevention and for treating severe cases belongs to the specific competence of the health sector. In the second line of action, the aim is to intervene before reserves are depleted. Here, primary prevention and health promotion are choice activities. This frame of reference will be used for analysing existing health programs for preschool children and how they propose to improve the management of malnutrition. This analysis will show that primary health care services can play a much more important role than usually attributed to them. Identifying these gaps and elaborating alternatives is the purpose of this article. PMID- 11943645 TI - [Choice of nutritional interventions in Congo: an approach for primary health care centres and for the community]. AB - This research is aimed at testing and adapting a comprehensive and participatory approach to identification, selection and evaluation of nutritional interventions in Congo. The mains tools utilized here are a causal model, a ranking table and a HIPPOPOC table. The causal model was built by a multi-sectional team of 35 persons and used to conceptualize the nutrition of young children in Congo, to identify vulnerable determinants and relevant interventions. Three main determinants were identified: i) Food consumption of the child; ii) Health status of the child and iii) Caring practices of children and mothers. Each determinant was developed into a sub-model. For each sub-model, the team identified vulnerable factors and relevant interventions. The identification of six interventions was based on experience, scientific knowledge and on-going interventions of the members of the group during participatory discussions. As resources were limited, it was not possible to implement all of the six interventions; thus a selection of the most appropriate interventions was made using a ranking table. Before building the ranking table, a choice of the most appropriate criteria was made by the group: technical feasibility, acceptability by the population, acceptability by health workers, financial feasibility, short term impact, potential of participation and sustainability. The ranking table for intervention selection allows a critical discussion on each intervention and leads to prioritization. Once the table was completed, the three most appropriate interventions were selected: communication for behavior change, improvement of quality of care, and promotion of improved complementary foodstuff production. Two of the interventions (psychosocial stimulation and community-based) identified needed more research for implementation. In order to determine clear objectives for intervention and to set up a monitoring and evaluation system, a practical tool was used: the HIPPOPOC table. This table represents a simplified view of the intervention in which project inputs (resources), processes (activities), outputs (immediate results), outcomes (short- or mid-term effects) and impacts (long-term effects) are clearly defined. The approach used here is based firstly on completeness as a way of guaranteeing the success of the design, implementation and evaluation of an intervention. Completeness means that all factors that may affect actions and their impact are taken into account. Secondly, participation is a key element of the process. All actors playing an active role in the decision-making process are present during the planning and evaluation process. This approach is meant to empower individuals and the community as a whole and develop within the community an awareness and a competence for problem solving. It involves multiple stakeholders (target groups, supervisors, sponsors, central as well as peripheral workers, experts, etc.) in design, implementation, needs assessment, monitoring and evaluation. PMID- 11943647 TI - [Nutrition surveillance: 25 years later]. AB - Nutrition surveillance is a rather recent application of epidemiological surveillance to nutrition. It covers not only the morbidity and mortality of nutritional disorders, but is at least as much interested in their major determinants. After some fifteen years of trial and error, nutrition surveillance has today found its way: its concepts, methods and uses are well established by now. After retrieving experiences conducted since the seventies and the eighties, and on the basis of their own experience, the authors have come to identify a few major features of nutrition surveillance: predominance of the supply of information over the collection of data; the early identification of users; the importance of causal analysis as a departure point for conceiving a surveillance activity; the highly selective choice of data to be collected; a permanent concern for sustainability; the participation of all actors (or "stakeholders") and, from a technical viewpoint, the place of qualitative data collection techniques (borrowed from social anthropology) to complement quantitative data. The authors summarize the major steps followed in the design and implementation of a nutrition surveillance activity, emphasizing the benefits of a careful and in-depth preparation during the so-called "pre-surveillance" phase. PMID- 11943646 TI - [At the start of the 21st century, we have the means to reduce malnutrition in sub-Saharan Africa, but will we do it?]. AB - The number of malnourished children in Africa continues to increase. It has been proven that such tendency could be stopped or even reversed if the lessons learnt from successful community nutrition programs were applied. The authors do not deny the role that poor socio-economic conditions play on malnutrition, and that those conditions need to be improved for a long-lasting impact, but they also argue that it is possible to act in parallel and get positive results, without waiting for macro-economic improvements. Successful projects such as those from Tamil Nadu in India, Iringa in Tanzania, Secaline in Madagacar, the PNC in Senegal, the AIN-C in Honduras, Prosen in Cameroon all applied the same principles and a similar approach. To reach success, international nutrition experts identified eighteen principles which should be used when designing and monitoring a community nutrition program, and there are also five phases to follow during the design stage. Finally, the authors suggest that if nutrition seems to often be underfunded, it is mainly because nutritionists in general have not been able to come up with feasible and well-conceived programmes. A few ideas are provided on how to access financing from health sector programs, by ensuring that the community nutrition programme helps implement certain aspects of those health sector programs. The conclusion is that there is no excuse not to start improving the nutritional status of African children now. PMID- 11943648 TI - Combining Coronary Revascularization and Valvular Surgery on a Beating Heart: An Analysis. PMID- 11943649 TI - Straining to understand mechanotransduction in the lung. PMID- 11943651 TI - Ventilation-induced lung injury and mechanotransduction: stretching it too far? AB - The Acute Respiratory Distress Syndrome Network clinical trial on ventilation of critically ill patients has drawn attention to the potential side effects of mechanical ventilation. Both clinical and basic research have demonstrated that injurious ventilation strategies can initiate or perpetuate local and systemic inflammatory responses. There are four principal mechanisms that can produce such a response. 1) Ventilation, especially with high ventilation pressures and zero positive end-expiratory pressure, can cause stress failure of the plasma membrane and of epithelial and endothelial barriers. Stress failure of the plasma membrane causes necrosis, which leads to liberation of both preformed inflammatory mediators and agents that stimulate other cells that are still intact to produce such mediators. 2) Stress failure of the barriers causes loss of compartmentalization with spread of mediators and bacteria throughout the body as a consequence. 3) Less injurious ventilation strategies that do not cause tissue destruction can elicit release of mediators by more specific mechanisms, presumably through activation of stretch-activated signaling cascades (mechanotransduction). 4) Ventilation with increasing positive pressures raises the pressure in the pulmonary circulation and thus vascular shear stress, both of which are known stimuli for endothelial cells. These different mechanisms should be taken into account in the design and the interpretation of studies on molecular mechanisms of ventilation-induced lung injury. PMID- 11943650 TI - Cyclic stretch activates ERK1/2 via G proteins and EGFR in alveolar epithelial cells. AB - Mechanical stimuli are transduced into intracellular signals in lung alveolar epithelial cells (AEC). We studied whether mitogen-activated protein kinase (MAPK) pathways are activated during cyclic stretch of AEC. Cyclic stretch induced a rapid (within 5 min) increase in extracellular signal-regulated kinase 1/2 (ERK1/2) phosphorylation in AEC. The inhibition of Na(+), L-type Ca(2+) and stretch-activated ion channels with amiloride, nifedipine, and gadolinium did not prevent the stretch-induced ERK1/2 activation. The inhibition of Grb2-SOS interaction with an SH3 binding sequence peptide, Ras with a farnesyl transferase inhibitor, and Raf-1 with forskolin did not affect the stretch-induced ERK1/2 phosphorylation. Moreover, cyclic stretch did not increase Ras activity, suggesting that stretch-induced ERK1/2 activation is independent of the classical receptor tyrosine kinase-MAPK pathway. Pertussis toxin and two specific epidermal growth factor receptor (EGFR) inhibitors (AG-1478 and PD-153035) prevented the stretch-induced ERK1/2 activation. Accordingly, in primary AEC, cyclic stretch activates ERK1/2 via G proteins and EGFR, in Na(+) and Ca(2+) influxes and Grb2 SOS-, Ras-, and Raf-1-independent pathways. PMID- 11943652 TI - Cyclic mechanical stretch induces VEGF and FGF-2 expression in pulmonary vascular smooth muscle cells. AB - Vascular endothelial growth factor (VEGF) and basic (b) fibroblast growth factor (FGF-2/bFGF) are involved in vascular development and angiogenesis. Pulmonary artery smooth muscle cells express VEGF and FGF-2 and are subjected to mechanical forces during pulsatile blood flow. The effect of stretch on growth factor expression in these cells is not well characterized. We investigated the effect of cyclic stretch on the expression of VEGF and FGF-2 in ovine pulmonary artery smooth muscle cells. Primary confluent cells from 6-wk-old lambs were cultured on flexible silicon membranes and subjected to cyclic biaxial stretch (1 Hz; 5-25% stretch; 4-48 h). Nonstretched cells served as controls. Expression of VEGF and FGF-2 was determined by Northern blot analysis. Cyclic stretch induced expression of both VEGF and FGF-2 mRNA in a time- and amplitude-dependent manner. Maximum expression was found at 24 h and 15% stretch (VEGF: 1.8-fold; FGF-2: 1.9-fold). These results demonstrate that mechanical stretch regulates VEGF and FGF-2 gene expression, which could play a role in pulmonary vascular development or in postnatal pulmonary artery function or disease. PMID- 11943653 TI - Bronchial epithelial compression regulates MAP kinase signaling and HB-EGF-like growth factor expression. AB - Airway smooth muscle constriction leads to the development of compressive stress on bronchial epithelial cells. Normal human bronchial epithelial cells exposed to an apical-to-basal transcellular pressure difference equivalent to the computed stress in the airway during bronchoconstriction demonstrate enhanced phosphorylation of extracellular signal-regulated kinase (ERK). The response is pressure dependent and rapid, with phosphorylation increasing 14-fold in 30 min, and selective, since p38 and c-Jun NH(2)-terminal kinase phosphorylation remains unchanged after pressure application. Transcellular pressure also elicits a ninefold increase in expression of mRNA encoding heparin-binding epidermal growth factor-like growth factor (HB-EGF) after 1 h, followed by prominent immunostaining for pro-HB-EGF after 6 h. Inhibition of the ERK pathway with PD 98059 results in a dose-dependent reduction in pressure-induced HB-EGF gene expression. The magnitude of the HB-EGF response to transcellular pressure and tumor necrosis factor (TNF)-alpha (1 ng/ml) is similar, and the combined mechanical and inflammatory stimulus is more effective than either stimulus alone. These results demonstrate that compressive stress is a selective and potent activator of signal transduction and gene expression in bronchial epithelial cells. PMID- 11943654 TI - Vascular regulation of type II cell exocytosis. AB - To determine whether lung capillary pressure regulates surfactant secretion, we viewed alveoli of the constantly inflated, isolated blood-perfused rat lung by fluorescence microscopy. By alveolar micropuncture we infused fura 2 and lamellar body (LB)-localizing dyes for fluorescence detection of, respectively, the alveolar cytosolic Ca(2+) concentration ([Ca(2+)](i)) and type II cell exocytosis. Increasing left atrial pressure (Pla) from 5 to 10 cmH(2)O increased septal capillary diameter by 26% and induced marked alveolar [Ca(2+)](i) oscillations that abated on relief of pressure elevation. The rate of loss of LB fluorescence that reflects the LB exocytosis rate increased fourfold after the pressure elevation and continued at the same rate even after pressure and [Ca(2+)](i) oscillations had returned to baseline. In alveoli pretreated with either 1,2-bis(2-aminophenoxy)ethane-N,N,N',N'-tetraacetic acid-AM, the intracellular Ca(2+) chelator, or heptanol, the gap junctional blocker, the pressure-induced exocytosis was completely inhibited. We conclude that capillary pressure and surfactant secretion are mechanically coupled. The secretion initiates in a Ca(2+)-dependent manner but is sustained by Ca(2+)-independent mechanisms. PMID- 11943655 TI - Pressure-induced endothelial Ca(2+) oscillations in lung capillaries. AB - Endothelial second messenger responses may contribute to the pathology of high vascular pressure but remain poorly understood because of the lack of direct in situ quantification. In lung venular capillaries, we determined endothelial cytosolic Ca(2+) concentration [Ca(2+)](i) by the fura 2 ratioing method. Pressure elevation increased mean endothelial [Ca(2+)](i) by Ca(2+) influx through gadolinium-inhibitable channels and amplified [Ca(2+)](i) oscillations by Ca(2+) release from intracellular stores. Endothelial [Ca(2+)](i) transients were induced by pressure elevations of as little as 5 cmH(2)O and increased linearly with higher pressures. Heptanol inhibition of [Ca(2+)](i) oscillations in a subset of endothelial cells indicated that oscillations originated from pacemaker endothelial cells and were propagated to adjacent nonpacemaker cells by gap junctional communication. Our findings indicate the presence of a sensitive, active endothelial response to pressure challenge in lung venular capillaries that may be relevant in the pathogenesis of pressure-induced lung microvascular injury. PMID- 11943657 TI - Nitric oxide production by human macrophages: there's NO doubt about it. PMID- 11943656 TI - Keratinocyte and hepatocyte growth factors in the lung: roles in lung development, inflammation, and repair. AB - A growing body of evidence indicates that the epithelial-specific growth factors keratinocyte growth factor (KGF), fibroblast growth factor (FGF)-10, and hepatocyte growth factor (HGF) play important roles in lung development, lung inflammation, and repair. The therapeutic potential of these growth factors in lung disease has yet to be fully explored. KGF has been best studied and has impressive protective effects against a wide variety of injurious stimuli when given as a pretreatment in animal models. Whether this protective effect could translate to a treatment effect in humans with acute lung injury needs to be investigated. FGF-10 and HGF may also have therapeutic potential, but more extensive studies in animal models are needed. Because HGF lacks true epithelial specificity, it may have less potential than KGF and FGF-10 as a targeted therapy to facilitate lung epithelial repair. Regardless of their therapeutic potential, studies of the unique roles played by these growth factors in the pathogenesis and the resolution of acute lung injury and other lung diseases will continue to enhance our understanding of the complex pathophysiology of inflammation and repair in the lung. PMID- 11943658 TI - Killing of Klebsiella pneumoniae by human alveolar macrophages. AB - We investigated putative mechanisms by which human surfactant protein A (SP-A) effects killing of Klebsiella pneumoniae by human alveolar macrophages (AMs) isolated from bronchoalveolar lavagates of patients with transplanted lungs. Coincubation of AMs with human SP-A (25 microg/ml) and Klebsiella resulted in a 68% decrease in total colony forming units by 120 min compared with AMs infected with Klebsiella in the absence of SP-A, and this SP-A-mediated effect was abolished by preincubation with N(G)-monomethyl-L-arginine. Incubation of transplant AMs with SP-A increased intracellular Ca(2+) concentration ([Ca(2+)](i)) by 70% and nitrite and nitrate (NO(x)) production by 45% (from 0.24 +/- 0.02 to 1.3 +/- 0.21 nmol small middle dot 10(6) AMs(-1).h(-1)). Preincubation with 1,2-bis(2-aminophenoxy)ethane-N,N,N',N'-tetraacetic acid acetoxymethyl ester inhibited the increase in [Ca(2+)](i) and abrogated the SP-A mediated Klebsiella phagocytosis and killing. In contrast, incubation of AMs from normal volunteers with SP-A decreased both [Ca(2+)](i) and NO(x) production and did not result in killing of Klebsiella. Significant killing of Klebsiella was also seen in a cell-free system by sustained production of peroxynitrite (>1 microM/min) at pH 5 but not at pH 7.4. These findings indicate that SP-A mediates pathogen killing by AMs from transplant lungs by stimulating phagocytosis and production of reactive oxygen-nitrogen intermediates. PMID- 11943659 TI - The time has finally arrived: use of intravital microscopy in the airway circulation. PMID- 11943660 TI - Leukocyte recruitment in the airways: an intravital microscopic study of rat tracheal microcirculation. AB - Because of its relative inaccessibility, inflammatory cell extravasation within the airway circulation in vivo has been difficult to investigate in real time. A new method has been established using intravital microscopy in the anesthetized rat to visualize leukocytes in superficial postcapillary venules of the trachea. This technique has been validated using local superfusion of lipopolysaccharide (LPS) and N-formyl-methionyl-leucyl-phenylalanine (FMLP). Basal leukocyte rolling velocity (55.4 +/- 9.3 microm/s) and adhesion (1.4 +/- 0.3 cells/100 microm) were monitored in postcapillary venules (33.9 +/- 1.3 microm diameter). At all time points up to 90 min, these parameters were unaltered in control rats (n = 7). In contrast, vessels exposed to 1 microg/ml of LPS (n = 6) exhibited a 57% reduction in leukocyte rolling velocity and an increase in the number of adherent cells (4.7 +/- 1 cells/100 microm, P < 0.05). Superfusion with 0.1 microM of FMLP (n = 6) also resulted in a 45% reduction in rolling velocity and an increase in adherent cells (4 +/- 0.7 cells/100 microm, P < 0.05). Histological evaluation confirmed local stimulus-induced leukocyte extravasation. These results demonstrate leukocyte recruitment in the airway microvasculature and provide an important new method to study airway inflammation in real time. PMID- 11943662 TI - Selective expansion of fibroblast subpopulations from pulmonary artery adventitia in response to hypoxia. AB - Proliferation of fibroblasts contributes to the adventitial thickening observed during the development of hypoxia-induced pulmonary hypertension. However, whether all or only specific subpopulations of fibroblasts proliferate during this process is unknown. Because lung, skin, and gingiva contain multiple fibroblast subpopulations, we hypothesized that the pulmonary artery (PA) adventitia of neonatal calves is composed of multiple fibroblast subpopulations and that only selective subpopulations expand under chronic hypoxic conditions. Fibroblast subpopulations were isolated from PA adventitia of control calves using limited dilution cloning techniques. These subpopulations exhibited marked differences in morphology, actin expression, and serum-stimulated growth. Only select fibroblast subpopulations demonstrated the ability to proliferate in response to hypoxia. Fibroblast subpopulations were similarly isolated from calves exposed to hypoxia (14 days). With regard to morphology, actin expression, and serum-stimulated growth of subpopulations, there were no obvious differences in fibroblast subpopulations between the hypoxic and the control calves. However, the number of fibroblast subpopulations with about a twofold increase in hypoxia induced DNA synthesis was significantly greater in the hypoxic calves (26%) compared with control calves (10%). We conclude that the bovine PA adventitia comprises numerous phenotypically and biochemically distinct fibroblast subpopulations and that select subpopulations expand in response to chronic hypoxia. PMID- 11943661 TI - Activation of NF-kappaB-dependent gene expression by silica in lungs of luciferase reporter mice. AB - Occupational exposure to crystalline silica is associated with the development of pulmonary inflammation and silicosis, yet how silica initiates pulmonary fibrosis and which cell types are involved are unclear. In studies here, we hypothesized that silica particles interact initially with pulmonary epithelial cells and alveolar macrophages (AMs) to cause transcriptional activation of nuclear factor (NF)-kappaB-regulated genes encoding inflammatory cytokines. Exposure of NF kappaB luciferase reporter mice intratracheally to silica or lipopolysaccharide (LPS), but not the nonfibrogenic particle titanium dioxide (TiO(2)), increased immunoreactivity of luciferase protein in bronchiolar epithelial cells and AMs. Ribonuclease protection assays revealed significant (P < or = 0.05) increases in mRNA levels of inducible nitric oxide synthase, tumor necrosis factor-alpha, macrophage inflammatory protein-2, macrophage chemotactic protein-1 (MCP-1), interferon-gamma, interleukin (IL)-6, and IL-12 in lung homogenates of reporter mice after exposures to silica or LPS. Immunoreactivity of MCP-1 in these animals was localized to AMs and epithelial cells. These data are the first to show activation of NF-kappaB in situ by fibrogenic particles in pulmonary epithelial cells and AMs. Increased expression of NF-kappaB-related inflammatory cytokines by these cell types, which first encounter silica after inhalation, may be critical to the initiation of silica-associated lung diseases, thus providing a rationale for focusing on NF-kappaB in preventive and therapeutic strategies. PMID- 11943663 TI - Iron uptake and Nramp2/DMT1/DCT1 in human bronchial epithelial cells. AB - The capacity of natural resistance-associated macrophage protein-2 [Nramp2; also called divalent metal transporter-1 (DMT1) and divalent cation transporter-1 (DCT1)] to transport iron and its ubiquitous expression make it a likely candidate for transferrin-independent uptake of iron in peripheral tissues. We tested the hypothesis that non-transferrin-bound iron uptake by airway epithelial cells is associated with Nramp2/DMT1/DCT1 and that exposure to iron can increase Nramp2/DMT1/DCT1 mRNA and protein expression and transport of this metal. Exposure of BEAS-2B cells to ferric ammonium citrate (FAC) resulted in a decrease in Fe(3+) concentration in the supernatant that was dependent on time and initial iron concentration. In the presence of internalized calcein, FAC quenched the fluorescent signal, indicating intracellular transport of the metal. The Nramp2/DMT1/DCT1 mRNA isoform without an iron-response element (IRE) increased with exposure of BEAS-2B cells to FAC. RT-PCR demonstrated no change in the mRNA for the isoform with an IRE. Similarly, Western blot analysis for the isoform without an IRE confirmed an increased expression of this protein after FAC exposure, whereas the isoform with an IRE exhibited no change. Finally, immunohistochemistry revealed an increase in the isoform without an IRE in the rat lung epithelium after instillation of FAC. Comparable to mRNA and protein increases, iron transport was elevated after pretreatment of BEAS-2B cells with iron-containing compounds. We conclude that airway epithelial cells increase mRNA and expression of the Nramp2/DMT1/DCT1 without an IRE after exposure to iron. The increase results in an elevated transport of iron and its probable detoxification by these cells. PMID- 11943664 TI - Identification of protein disulfide isomerase as an endothelial hypoxic stress protein. AB - Endothelial cells (EC) exposed to hypoxia upregulate a unique set of five stress proteins. These proteins are upregulated in human and bovine aortic and pulmonary artery EC and are distinct from heat shock or glucose-regulated proteins. We previously identified two of these proteins as the glycolytic enzymes glyceraldehyde-3-phosphate dehydrogenase and enolase and postulated that the remaining proteins were also glycolytic enzymes. Using SDS-PAGE, tryptic digestion, and NH(2)-terminal amino acid sequencing, we report here the identification of the 56-kDa protein as protein disulfide isomerase (PDI). PDI is upregulated by hypoxia at the mRNA level and follows a time course similar to that of the protein, with maximal upregulation detected after exposure to 18 h of 0% O(2). Neither smooth muscle cells nor fibroblasts upregulate PDI to the same extent as EC, which correlates with their decreased hypoxia tolerance. Upregulation of PDI specifically in EC may contribute to their ability to tolerate hypoxia and may occur through PDI's functions as a prolyl hydroxylase subunit, protein folding catalyst, or molecular chaperone. PMID- 11943665 TI - Laminin alpha-chain expression and basement membrane formation by MLE-15 respiratory epithelial cells. AB - Basement membranes have a critical role in alveolar structure and function. Alveolar type II cells make basement membrane constituents, including laminin, but relatively little is known about the production of basement membrane proteins by murine alveolar type II cells and a convenient system is not available to study basement membrane production by murine alveolar type II cells. To facilitate study of basement membrane production, with particular focus on laminin chains, we examined transformed murine distal respiratory epithelial cells (MLE-15), which have many structural and biochemical features of alveolar type II cells. We found that MLE-15 cells produce laminin-alpha5, a trace amount of laminin-alpha3, laminins-beta1 and -gamma1, type IV collagen, and perlecan. Transforming growth factor-beta1 significantly induces expression of laminin alpha1. When grown on a fibroblast-embedded collagen gel, MLE-15 cells assemble a basement membrane-like layer containing laminin-alpha5. These findings indicate that MLE-15 cells will be useful in modeling basement membrane production and assembly by alveolar type II cells. PMID- 11943666 TI - Fusion of lung lobes and vessels in mouse embryos heterozygous for the forkhead box f1 targeted allele. AB - Previously, we showed that newborn forkhead box (Fox)f1(+/-) mice with diminished pulmonary FoxF1 levels died of severe lung hemorrhage and exhibited abnormal formation of alveolar sacs and capillaries. Another group recently reported that Foxf1(+/-) mouse embryos displayed a number of organ and skeletal defects including fusion of lung lobes. However, identification of pulmonary genes whose altered expression was associated with the lobular fusion defect in Foxf1(+/-) lungs remains uncharacterized. The present study was conducted to determine the nature of the malformations leading to lung fusions in the FoxF1 embryos and to identify potential signaling pathways influenced by FoxF1 haploinsufficiency. We show that Foxf1(+/-) embryos exhibit defects in formation and branching of primary lung buds, which causes fusion of the right lung lobes and vessels. The severity of the Foxf1(+/-) lung fusions was correlated with decreased levels of FoxF1 mRNA. In situ hybridization studies demonstrated that the defective primary lung-bud development in early Foxf1(+/-) embryos was associated with fewer pulmonary mesenchymal-epithelial interfaces. Defects in branching morphogenesis in the Foxf1(+/-) embryos were associated with altered expression of the fibroblast growth factor-10, bone morphogenetic protein-4, and the Gli3 transcription factor, which are known to influence primary lung-bud development. PMID- 11943667 TI - Feeble bronchomotor responses in diabetic rats in association with decreased sensory neuropeptide release. AB - Type I diabetes is associated with a low incidence of asthma. We tested whether a decrease in sensory neuropeptide release is associated with an attenuated bronchoconstrictive response to field stimulation (FS; 100 stimuli, 20 V, 0.1 ms, 20 Hz) in streptozotocin (STZ)-induced diabetes. The organ fluid of the preparations were also tested for substance P, calcitonin gene-related peptide (CGRP), and somatostatin concentrations by RIA. Preparations were from either normal rats or those pretreated with 50 mg/kg STZ iv 8 wk before experiment. A group of STZ-treated animals was supplied with insulin delivery (4 IU/day sc) implants between 4 and 8 wk. A subgroup was formed to study the effect of capsaicin desensitization. The atropine-resistant contraction was attenuated by diabetes without capsaicin-sensitive relaxation response. Exogenous CGRP and substance P potentiated, whereas somatostatin inhibited (1 nM-10 microM) the FS induced contractions in rings from either group. FS released somatostatin, CGRP, and substance P from 0.17 +/- 0.024, 0.15 +/- 0.022, and 1.65 +/- 0.093 to 0.58 +/- 0.032, 0.74 +/- 0.122, and 5.34 +/- 0.295 in preparations from normal, and from 0.19 +/- 0.016, 0.11 +/- 0.019, and 0.98 +/- 0.116 to 0.22 +/- 0.076, 0.34 +/- 0.099, and 1.84 +/- 0.316 fmol/mg wet wt in preparations from diabetic rats. Insulin supplementation restored neuropeptide release in rings from STZ-treated rats. The results show that the decreased FS-induced contractions occurred with a decrease in sensory neuropeptide release in STZ-diabetic rats. PMID- 11943668 TI - Vitamin A deficiency promotes bronchial hyperreactivity in rats by altering muscarinic M(2) receptor function. AB - Vitamin A deficiency (VAD) remains an important health problem among children in developing countries. Children living in these areas have a higher mortality from respiratory infections, which likely results in part from suboptimal nutrition, including VAD. Bronchial hyperreactivity can follow viral respiratory infections and may complicate the recovery. To investigate whether VAD promotes bronchial hyperreactivity, we have assessed methacholine-induced bronchoconstriction in VAD and vitamin A-sufficient rats. Bronchial constriction developed at lower concentrations of inhaled methacholine in VAD than in vitamin A-sufficient rats. This did not result from an increase in the bronchial wall thickness or the clearance of a small molecule (with a size similar to methacholine) from the air space. The function and abundance of the muscarinic M(2) receptors in bronchial tissue were reduced in VAD rats, suggesting that this receptor may contribute to these animals' diminished ability to limit cholinergic-mediated bronchoconstriction. A similar reduction in muscarinic M(2) receptor function has been observed in asthma. Vitamin A (retinol) and its congeners (retinoids) may be required to regulate bronchial responsiveness in addition to maintaining a normal bronchial epithelium. PMID- 11943669 TI - Role of Ras in metal-induced EGF receptor signaling and NF-kappaB activation in human airway epithelial cells. AB - We showed previously that epithelial growth factor (EGF) receptor (EGFR) signaling is triggered by metallic compounds associated with ambient air particles. Specifically, we demonstrated that As, Zn, and V activated the EGFR tyrosine kinase and the downstream kinases MEK1/2 and ERK1/2. In this study, we examined the role of Ras in EGFR signaling and the nuclear factor-kappaB (NF kappaB) activation pathway and the possible interaction between these two signaling pathways in a human airway epithelial cell line (BEAS-2B) exposed to As, V, or Zn ions. Each metal significantly increased Ras activity, and this effect was inhibited by the EGFR tyrosine kinase activity inhibitor PD-153035. Adenoviral-mediated overexpression of a dominant-negative mutant form of Ras(N17) significantly blocked MEK1/2 or ERK1/2 phosphorylation in As-, Zn-, or V-exposed BEAS-2B cells but caused little inhibition of V-, Zn- or EGF-induced EGFR tyrosine phosphorylation. This confirmed Ras as an important intermediate effector in EGFR signaling. Interestingly, V, but not As, Zn, or EGF, induced IkappaBalpha serine phosphorylation, IkappaBalpha breakdown, and NF-kappaB DNA binding. Moreover, PD-153035 and overexpression of Ras(N17) each significantly blocked V-induced IkappaBalpha breakdown and NF-kappaB activation, while inhibition of MEK activity with PD-98059 failed to do so. In summary, exposure to As, Zn, and V initiated EGFR signaling and Ras-dependent activation of MEK1/2 and ERK1/2, but only V induced Ras-dependent NF-kappaB nuclear translocation. EGFR signaling appears to cross talk with NF-kappaB signaling at the level of Ras, but additional signals appear necessary for NF-kappaB activation. Together, these data suggest that, in V-treated BEAS-2B cells, Ras-dependent signaling is essential, but not sufficient, for activation of NF-kappaB. PMID- 11943670 TI - Th2 cytokine regulation of type I collagen gel contraction mediated by human lung mesenchymal cells. AB - Asthma is characterized by chronic inflammation of the airway wall with the presence of activated T helper 2 (Th2) lymphocytes. The current study assessed the ability of Th2 cytokines to modulate fibroblast-mediated contraction of collagen gels to determine if Th2 cytokines could contribute to tissue remodeling by altering mesenchymal cell contraction. Human fetal lung fibroblasts, human adult bronchial fibroblasts and human airway smooth muscle cells were cast into native type I collagen gels and allowed to contract in the presence or absence of IL (interleukin)-4, IL-5, IL-10, or IL-13. IL-4 and IL-13 but not IL-5 and IL-10 augmented collagen gel contraction in a concentration-dependent manner. Neither IL-4 nor IL-13 altered fibroblast production of transforming growth factor-beta or fibronectin. Both, however, decreased fibroblast prostaglandin (PG) E(2) release. Decreased PGE(2) release was associated with a decreased expression of cyclooxygenase 1 and 2 protein and mRNA. Indomethacin completely inhibited PGE(2) release and also augmented contraction. IL-4 and IL-13, however, added together with indomethacin further augmented contraction suggesting both a PGE-dependent and a PGE-independent effect. These findings suggest that IL-4 and IL-13 may modulate airway tissue remodeling and, therefore, could play a role in the altered airway connective tissue which characterizes asthma. PMID- 11943671 TI - Pulmonary vasoconstriction by serotonin is inhibited by S-nitrosoglutathione. AB - Nitric oxide (NO) functions as an endothelium-derived relaxing factor by activating guanylate cyclase to increase cGMP levels. However, NO and related species may also regulate vascular tone by cGMP-independent mechanisms. We hypothesized that naturally occurring NO donors could decrease the pulmonary vascular response to serotonin (5-HT) in the intact lung through chemical interactions with 5-HT(2) receptors. In isolated rabbit lung preparations and isolated pulmonary artery (PA) rings, 50-250 microM S-nitrosoglutathione (GSNO) inhibited the response to 0.01-10 microM 5-HT. The vasoconstrictor response to 5 HT was mediated by 5-HT(2) receptors in the lung, since it could be blocked completely by the selective inhibitor ketanserin (10 microM). GSNO inhibited the response to 5-HT by 77% in intact lung and 82% in PA rings. In PA rings, inhibition by GSNO could be reversed by treatment with the thiol reductant dithiothreitol (10 mM). 3-Morpholinosydnonimine (100-500 microM), which releases NO and O simultaneously, also blocked the response to 5-HT. Its chemical effects, however, were distinct from those of GSNO, because 5-HT-mediated vasoconstriction was not restored in isolated rings by dithiothreitol. In the intact lung, neither NO donor altered the vascular response to endothelin, which activates the same second-messenger vasoconstrictor system as 5-HT. These findings, which did not depend on guanylate cyclase, are consistent with chemical modification by NO of the 5-HT(2) G protein-coupled receptor system to inhibit vasoconstriction, possibly by S-nitrosylation of the receptor or a related protein. This study demonstrates that GSNO can regulate vascular tone in the intact lung by a reversible mechanism involving inhibition of the response to 5-HT. PMID- 11943672 TI - CD4(+) T cell-dependent airway mucus production occurs in response to IL-5 expression in lung. AB - The potential role of airway interleukin-5 (IL-5) expression in eliciting mucus production was demonstrated in a pulmonary IL-5 transgenic mouse model (NJ.1726) in which naive transgenic mice display comparable levels of airway mucus relative to allergen-sensitized and -challenged wild-type mice. The intrinsic mucus accumulation of NJ.1726 was abolished in compound transgenic-gene knockout mice deficient of either CD4(+) cells [NJ.1726/CD4(-/-)] or alphabeta T cell receptor positive (TCR(+)) cells [NJ.1726/alphabeta TCR(-/-)]. In addition, mucus production in naive NJ.1726 was inhibited by >90% after administration of the soluble anti-IL-4 receptor alpha-subunit antagonist. The loss of mucus production in NJ.1726/CD4(-/-), NJ.1726/alphabeta TCR(-/-), and anti-IL-4 receptor alpha subunit antagonist-treated mice occurred notwithstanding the significant pulmonary eosinophilia and expansion of airway B cells induced by ectopic IL-5 expression. Furthermore, the loss of mucus accumulation occurred in these mice despite elevated levels of airway and peripheral IL-5, indicating that IL-5 does not directly induce goblet cell metaplasia and mucus production. Thus pulmonary expression of IL-5 alone is capable of inducing CD4(+) T cell-dependent goblet cell metaplasia, apparently mediated by IL-4 receptor alpha-subunit-ligand interactions, and represents a previously unrecognized novel pathway for augmenting allergen-induced mucus production. PMID- 11943673 TI - Diesel particles increase phosphatidylcholine release through a NO pathway in alveolar type II cells. AB - Diesel exhaust particles (DEPs) have been shown in vivo as well as in vitro to affect the respiratory function and in particular the immune response to infection and allergens. In the current study, we investigated the effect of DEPs on the production of phosphatidylcholine (PC), a major constituent of surfactant, by rat alveolar type II (ATII) primary cells in vitro. Our results demonstrate that incubation of ATII cells with DEPs lead to a time- and dose-dependent increase in labeled PC release. This effect was mimicked by nitric oxide (NO) donors and cGMP and was abolished by inhibitors of NO synthase (NOS). In addition, a NOS inhibitor inhibits by itself the basal secretion of PC. We next examined the effects of DEPs on NOS gene expression and showed that DEPs increase NO production and upregulate both protein content and mRNA levels of the inducible NOS (NOS II). Together our data demonstrate that DEPs alter the production of surfactant by ATII cells through a NO-dependent signaling pathway. PMID- 11943674 TI - Liposomal NAD(+) prevents diminished O(2) consumption by immunostimulated Caco-2 cells. AB - Accumulating data support the view that sepsis is associated with an acquired intrinsic derangement in the ability of cells to consume O(2), a phenomenon that has been termed "cytopathic hypoxia." We sought to use an in vitro "reductionist" model system using cultured cells stimulated with proinflammatory cytokines to test the hypothesis that cytopathic hypoxia is mediated, at least in part, by depletion of intracellular levels of NAD(+)/NADH secondary to activation of the nuclear enzyme poly(ADP-ribose) polymerase (PARP). We measured O(2) consumption by Caco-2 enterocytes growing on microcarrier beads after cells were incubated for 24 h under control conditions or with cytomix, a mixture of tumor necrosis factor-alpha, interleukin-1beta, and interferon-gamma. Immunostimulated cells consumed O(2) at about one-half the rate of control cells, but this effect was largely prevented if any one of the following pharmacological agents was present during the period of incubation with cytomix: 4,5-dihydroxy-1,3-benzene disulfonic acid, a superoxide radical anion scavenger; 2-(4-carboxyphenyl) 4,4,5,5-tetramethylimidazoline-1-oxyl-3-oxide, a nitric oxide scavenger; 5,10,15,20- tetrakis-[4-sulfonatophenyl]-porphyrinato-iron[III], a peroxynitrite (ONOO(-)) decomposition catalyst; urate, an ONOO(-) scavenger; 3-aminobenzamide, a PARP inhibitor; or N-(6-oxo-5,6-dihydrophenanthridin-2-yl)-N,N dimethylacetamide HCl, a chemically dissimilar and more potent PARP inhibitor. The decrease in O(2) uptake induced by cytomix was associated with decreased cellular levels of NAD(+)/NADH. The decrease in cellular NAD(+)/NADH content and the decrease in O(2) uptake induced by cytomix were completely abrogated if liposome-encapsulated NAD(+) was added to the cultures during immunostimulation. Empty liposomes also increased O(2) uptake by immunostimulated Caco-2 cells, but much less effectively than liposomes containing NAD(+). These data are consistent with the view that enterocytes exposed to proinflammatory cytokines consume less O(2) due to NAD(+)/NADH depletion secondary to activation of PARP by ONOO(-) or other oxidants. PMID- 11943675 TI - Activity of pulmonary edema fluid interleukin-8 bound to alpha(2)-macroglobulin in patients with acute lung injury. AB - The formation of alpha(2)-macroglobulin (alpha(2)-M)/interleukin-8 (IL-8) complexes may influence the biological activity of IL-8 and the quantitative assessment of IL-8 activity. Therefore, in this study, concentrations of free IL 8 and IL-8 complexes with alpha(2)-M were measured in pulmonary edema fluid samples from patients with acute lung injury/acute respiratory distress syndrome (ALI/ARDS) and compared with control patients with hydrostatic pulmonary edema. Patients with ALI/ARDS had significantly higher concentrations of alpha(2)-M (P < 0.01) as well as alpha(2)-M/IL-8 complexes (P < 0.05). Because a substantial amount of IL-8 is complexed to alpha(2)-M, standard assays of free IL-8 may significantly underestimate the concentration of biologically active IL-8 in the distal air spaces of patients with ALI/ARDS. Furthermore, IL-8 bound to alpha(2) M retained its biological activity, and this fraction of IL-8 was protected from proteolytic degradation. Thus complex formation may modulate the acute inflammatory process in the lung. PMID- 11943676 TI - Dopamine activates ERKs in alveolar epithelial cells via Ras-PKC-dependent and Grb2/Sos-independent mechanisms. AB - Recently it has been described that dopamine (DA), via dopaminergic type 2 receptors (D(2)R), activates the mitogen-activated protein kinase extracellular signal-regulated kinase (MAPK/ERK) proteins in alveolar epithelial cells (AEC), which results in the upregulation of Na(+)-K(+)-ATPase. In the present report, we used AEC to investigate the signaling pathway that links DA with ERK activation. Incubation of AEC with DA resulted in rapid and transient stimulation of ERK activity, which was mediated by Ras proteins and the serine/threonine kinase Raf 1. Pretreatment of AEC with Src homology 3 binding peptide, which blocks the interaction between Grb2 and Sos, did not prevent DA activation of ERK. Diacylglycerol (DAG)-dependent protein kinase C (PKC) isoenzymes, involved in the DA-mediated activation of ERK proteins as pretreatment with either bisindolylmaleimide or Ro-31-8220, prevented the phosphorylation of Elk-1, and quinpirole, a D(2)R activator, stimulates the translocation of PKCepsilon. Together, the data suggest that DA activated MAPK/ERK via Ras, Raf-1 kinase, and DAG-dependent PKC isoenzymes, but, importantly and contrary to the classical model, this pathway did not involve the Grb2-Sos complex formation. PMID- 11943677 TI - Activation of protein kinase A accelerates bovine bronchial epithelial cell migration. AB - Bronchial epithelial cell migration is required for the repair of damaged airway epithelium. We hypothesized that bronchial epithelial cell migration during wound repair is influenced by cAMP and the activity of its cyclic nucleotide-dependent protein kinase, protein kinase A (PKA). We found that, when confluent monolayers of bronchial epithelial cells are wounded, an increase in PKA activity occurs. Augmentation of PKA activity with a cell-permeable analog of cAMP, dibutyryl adenosine 3',5'-cyclic monophosphate, isoproterenol, or a phosphodiesterase inhibitor accelerated migration of normal bronchial epithelial cells in in vitro wound closure assays and Boyden chamber migration assays. A role for PKA activity was also confirmed with a PKA inhibitor, KT-5720, which reduced stimulated migration. Augmentation of PKA activity reduced the levels of active Rho and the formation of focal adhesions. These studies suggest that PKA activation modulates Rho activity, migration mechanisms, and thus bronchial epithelial repair mechanisms. PMID- 11943678 TI - Inhibition of the p38 MAP kinase pathway destabilizes smooth muscle length during physiological loading. AB - We tested the hypothesis that mechanical plasticity of airway smooth muscle may be mediated in part by the p38 mitogen-activated protein (MAP) kinase pathway. Bovine tracheal smooth muscle (TSM) strips were mounted in a muscle bath and set to their optimal length, where the active force was maximal (F(o)). Each strip was then contracted isotonically (at 0.32 F(o)) with ACh (maintained at 10(-4) M) and allowed to shorten for 180 min, by which time shortening was completed and the static equilibrium length was established. To simulate the action of breathing, we then superimposed on this steady distending force a sinusoidal force fluctuation with zero mean, at a frequency of 0.2 Hz, and measured incremental changes in muscle length. We found that TSM strips incubated in 10 microM SB-203580-HCl, an inhibitor of the p38 MAP kinase pathway, demonstrated a greater degree of fluctuation-driven lengthening than did control strips, and upon removal of the force fluctuations they remained at a greater length. We also found that the force fluctuations themselves activated the p38 MAP kinase pathway. These findings are consistent with the hypothesis that inhibition of the p38 MAP kinase pathway destabilizes muscle length during physiological loading. PMID- 11943679 TI - Gender differences in naphthalene metabolism and naphthalene-induced acute lung injury. AB - Humans are widely exposed to polycyclic aromatic hydrocarbons, commonly found in cigarette smoke and diesel exhaust. These can undergo site- and cell-specific metabolism to cytotoxic intermediates. Metabolism of naphthalene and Clara cell cytotoxicity have been extensively studied in male animals. To address whether male and female mice are equally susceptible to naphthalene, mice were injected with naphthalene, and lungs were examined 1, 2, 3, 6, and 24 h after treatment. By analysis of acute injury using differential permeability to fluorescent nuclear dyes and high-resolution histopathology, injury in female mice was found to be more extensive, occur earlier, and include permeable cells in proximal airways, including airway bifurcations. HPLC analysis of the products of cytochrome P-450 (CYP)-mediated metabolism in microdissected airways indicated that although both genders produced a predominance of products from CYP2F2, female mice produced more naphthalene dihydrodiol in distal airways, the primary sites of injury. We conclude that there are clear gender differences in susceptibility to naphthalene-induced injury and that differences in metabolism of naphthalene may play a role in elevated susceptibility in female mice. PMID- 11943680 TI - FAK blunts adenosine-homocysteine-induced endothelial cell apoptosis: requirement for PI 3-kinase. AB - Treatment of cultured bovine pulmonary endothelial cells (BPAEC) with adenosine (Ado) alone or in combination with homocysteine (Hc) leads to disruption of focal adhesion complexes, caspase-dependent degradation of components of focal adhesion complexes, and subsequent apoptosis. Endothelial cells transiently overexpressing paxillin or p130(Cas) cDNAs underwent Ado-Hc-induced apoptosis to an extent similar to that of cells transfected with vector alone. However, overexpression of focal adhesion kinase (FAK) cDNA blunted Ado-Hc-induced apoptosis. FAK constructs lacking the central catalytic domain or containing a point mutation, rendering the catalytic domain enzymatically inactive, did not provide protection from apoptosis. Constructs containing a mutation in the major autophosphorylation site (tyrosine-397) similarly did not prevent cell death. A FAK mutant in amino acid 395, deficient in phosphatidylinositol 3-kinase (PI 3-kinase) binding, was not able to blunt apoptosis. Finally, overexpression of FAK did not provide protection from apoptosis in the presence of LY-294002, a PI 3-kinase inhibitor. Taken together, these data suggest that the survival signals mediated by overexpression of FAK in response to Ado-Hc-induced apoptosis require a PI 3 kinase-dependent pathway. PMID- 11943681 TI - Leukotrienes mediate neurogenic inflammation in lungs of young rats infected with respiratory syncytial virus. AB - Respiratory syncytial virus (RSV) infection potentiates neurogenic inflammation in rat airways. Because some vascular effects of sensory nerves are mediated by cysteinyl leukotrienes (cysLTs), we studied whether the receptor antagonist montelukast inhibits neurogenic plasma extravasation in RSV-infected rats. Pathogen-free rats were inoculated at 2 wk (weanlings) or 12 wk (adults) of age with RSV or virus-free medium and treated with montelukast or its vehicle starting 1 day before inoculation. Five days postinoculation, we measured the extravasation of Evans blue-labeled albumin in the respiratory tract after stimulation of sensory nerves with capsaicin. Montelukast had no effect in the extrapulmonary airways but abolished albumin extravasation in the intrapulmonary airways of RSV-infected rats, with a larger effect in weanlings than in adults. Increased concentrations of 5-lipoxygenase-encoding mRNA and cysLTs, as well as numerous mast cells, were detected in the lung tissues of RSV-infected weanling rats. These observations suggest that the release of neuropeptides from capsaicin sensitive sensory nerves and nonneuronal cells in the lungs of RSV-infected young rats increases vascular permeability by promoting the release of leukotrienes from mast cells. PMID- 11943682 TI - Modulation of Na-K-2Cl cotransport by intracellular Cl(-) and protein kinase C delta in Calu-3 cells. AB - In this study, we tested the hypothesis that intracellular Cl(-) (Cl) regulates the activity of protein kinase C (PKC)-delta and thus the activation of Na-K-Cl cotransport (NKCC1) in a Calu-3 cell line. The alpha(1)-adrenergic agonist methoxamine (MOX) and hypertonic sucrose increased Cl and increased or decreased intracellular volume, respectively, without changing Cl concentration ([Cl( )](i)). Titration of [Cl(-)](i) from 20-140 mM in nystatin-permeabilized cell monolayers did not affect the baseline activity of PKC-delta, PKC-zeta, or rottlerin-sensitive NKCC1. At 200 mM Cl(-), rottlerin-sensitive NKCC1 was activated, and PKC isotypes were localized predominantly to a particulate fraction. MOX induced a biphasic increase in NKCC1 activity and PKC-delta in activity and particulate localization of PKC-delta and -zeta. Activity of NKCC1 and PKC-delta decreased with increasing Cl from 20 to 80 mM Cl then increased at 140-200 mM Cl apparently as an additive effect to high [Cl(-)](i) levels. Rottlerin inhibited the effects of MOX, which indicates that PKC-delta was required for activation of NKCC1. The results indicate that, in airway epithelial cells, a Cl electrochemical gradient alone is not sufficient to stimulate NKCC1 activity; rather, elevated activity of PKC-delta is necessary. Further, high Cl levels induce a subcellular redistribution of PKC-delta, which results in increased enzyme activity. PMID- 11943683 TI - Height in young adulthood and risk of death from cardiorespiratory disease: a prospective study of male former students of Glasgow University, Scotland. AB - To investigate the association between height in young, socially homogeneous males and cause-specific mortality, the authors conducted a prospective study of 8,361 male former students who underwent medical examinations while attending Glasgow University, Scotland, from 1948 to 1968. The mean age at examination was 20.5 (range, 16.1-30.0) years. The median follow-up time was 41.3 years. There were 863 deaths. In Cox proportional hazards modeling, there was no association between height and all-cause mortality with age-adjusted hazard ratios per 10-cm increase in height (hazard ratio = 0.92, 95% confidence interval: 0.83, 1.02). Height was inversely associated with all cardiovascular disease and coronary heart disease mortality, with hazard ratios per 10-cm increase in height of 0.78 (95% confidence interval: 0.66, 0.93) and 0.76 (95% confidence interval: 0.62, 0.93), respectively. Sizeable inverse associations with stroke and respiratory disease were also found, although these did not reach conventional levels of significance. There was no association with cancer or noncardiorespiratory disease mortality. There was a positive, although nonsignificant, association between height and mortality from aortic aneurysm. Controlling for confounding variables had little effect on these results. The findings suggest that factors operating in early life, and which influence height, also influence future cardiovascular health in men. PMID- 11943684 TI - Invited commentary: Height-cardiovascular disease relation: where to go from here? PMID- 11943685 TI - McCarron et al. respond to "height-cardiovascular disease relation": are all risk factors equal? PMID- 11943686 TI - Influence of retirement on leisure-time physical activity: the atherosclerosis risk in communities study. AB - Cross-sectional data suggest that leisure-time physical activity may increase during retirement. Prospective population-based studies are necessary to characterize leisure activity patterns through retirement among the same persons to verify this observation. Therefore, the purpose of this study was to describe the influence of retirement on leisure activity using data from Atherosclerosis Risk in Communities Study cohort participants aged 45-64 years at baseline. Physical activity was measured by the Baecke questionnaire in 1,825 African American and 5,957 White participants who were working at the initial visit (1986 1989) and either retired or working 6 years later (1993-1995). Participants who retired during follow-up were more likely to increase their sport participation and television watching than those who continued to work over the 6-year period. Among those reporting sport or exercise at baseline, those retiring over follow up were more likely to maintain their sport and exercise participation than those who continued to work across race-gender groups. Among those not reporting sport or exercise at baseline, those who retired were more likely to adopt activity than those who continued to work except for African-American women. In this study, retirement was associated with gains in sport and exercise participation as well as television watching. PMID- 11943687 TI - Social ties and change in social ties in relation to subsequent total and cause specific mortality and coronary heart disease incidence in men. AB - The authors prospectively examined the effects of social ties and change in social ties, as measured by a well-known social network index, on total and cause specific mortality and on coronary heart disease incidence in 28,369 US male health professionals aged 42-77 years in 1988. Over 10 years, the relative risk of total mortality for men in the lower two levels of social integration compared with more socially integrated men was 1.19 (95% confidence interval: 1.06, 1.34) after controlling for age, occupation, health behaviors, general physical condition, coronary risk factors, and dietary habits. In multivariate analysis, deaths from accidents and suicide and from other noncancer, noncardiovascular causes were significantly increased among less socially connected men. Socially isolated men also had an increased risk of fatal coronary heart disease (multivariate relative risk = 1.82, 95% confidence interval: 1.02, 3.23). An increase in the overall social network index between 1988 and 1996 was not significantly associated with subsequent 2-year mortality. In analyses of change in social network components restricted to older men, each categorical unit increase in number of close friends was significantly associated with a 29% decrease in risk of death. Increase in religious service attendance over time was also significantly predictive of decreased mortality. PMID- 11943688 TI - Estimating the effect of cardiovascular risk factors on all-cause mortality and incidence of coronary heart disease using G-estimation: the atherosclerosis risk in communities study. AB - Standard methods for analysis of cohort studies may give biased estimates of exposure effects in the presence of time-varying confounding. Such effects may instead be estimated by using G-estimation. This study aimed to examine the relations between important cardiovascular risk factors and all-cause mortality and risk of coronary heart disease (CHD), accounting for confounding between exposures over time using G-estimation. Results were compared with those from standard survival analyses (e.g., Weibull regression) with time-updated covariates. The dataset consisted of all participants in the Atherosclerosis Risk in Communities cohort study who had complete data on the first two of four visits, giving a sample of 13,898 people at baseline. Death and occurrence of CHD or stroke were recorded. G-estimated associations between several risk factors and mortality/CHD incidence differed from those estimated using standard survival analysis. The associations between mortality/CHD incidence and smoking, presence of diabetes, and use of antihypertensives were stronger than the standard survival estimates, while the G-estimated effect of low density lipoprotein and high density lipoprotein cholesterol on CHD incidence were more linear than the standard estimate. Complex relations between exposures over time may lead to biased exposure effect estimates in standard survival analyses. G-Estimation can be used to overcome such biases, and thus may have important implications for the analysis of observational studies. PMID- 11943689 TI - Chromosomal anomalies among the offspring of women with gestational diabetes. AB - A limited body of data over the past 35 years has suggested that autoimmunity may be responsible for some cases of aneuploidy. The role of diabetes mellitus in the etiology of chromosomal anomalies has been infrequently studied. This study was designed to compare the prevalence of chromosome abnormalities among the offspring of women with gestational diabetes and the offspring of women without it. The authors used data from 7,332 women who underwent amniocentesis in a prospective study of pregnancy outcome (1984-1988) and examined the prevalence of autosomal and sex chromosome defects associated with gestational diabetes. Among the offspring of 231 women with gestational diabetes, the crude prevalence of chromosomal defects was twice as high as that seen in the offspring of 7,101 women without gestational diabetes. These anomalies were predominantly numeric sex chromosome defects. After adjusting for potential confounding by maternal age, body mass index, education, and first-trimester exposures in multiple logistic regression analysis, the authors found that women with gestational diabetes were 7.7 times as likely (95% confidence interval: 2.8, 21.1) to have an infant with a numeric sex chromosome defect as those without gestational diabetes. These results support the theory that some women who develop gestational diabetes may have underlying biochemical changes that induce nondisjunction and the development of chromosomal defects. PMID- 11943690 TI - Blood transfusions as a risk factor for non-Hodgkin's lymphoma in the San Francisco Bay Area: a population-based study. AB - The incidence of non-Hodgkin's lymphoma (NHL) has risen dramatically over the past 50 years. In the search for new risk factors, blood transfusions have been investigated and shown to be associated with subsequent lymphoma in some studies. The authors tested this association in a population-based, case-control study conducted between 1988 and 1995 in the San Francisco Bay Area of California. A total of 1,591 histologically confirmed adult cases of NHL were included in this study. Cases were frequency matched to 2,515 control participants by sex, county of residence, and 5-year age intervals. Multivariate logistic regression models were used to examine the impact of possible confounders. Persons who reported a history of allogeneic transfusion were not at increased risk of NHL in this population (odds ratio (OR) = 1.0, 95% confidence interval (CI): 0.84, 1.2). No significant associations were seen when lymphomas were stratified by histologic subtype, grade, or latency period. However, autologous transfusions were associated with a decreased risk (OR = 0.39, 95% CI: 0.16, 0.94). In summary, these findings are similar to those of prior negative studies and do not support some previous reports of an adverse association between blood transfusion and NHL. PMID- 11943691 TI - Parkinson's disease risks associated with cigarette smoking, alcohol consumption, and caffeine intake. AB - A reduced risk for Parkinson's disease (PD) among cigarette smokers has been observed consistently during the past 30 years. Recent evidence suggests that caffeine may also be protective. Findings are presented regarding associations of PD with smoking, caffeine intake, and alcohol consumption from a case-control study conducted in western Washington State in 1992-2000. Incident PD cases (n = 210) and controls (n = 347), frequency matched on gender and age were identified from enrollees of the Group Health Cooperative health maintenance organization. Exposure data were obtained by in-person questionnaires. Ever having smoked cigarettes was associated with a reduced risk of PD (odds ratio (OR) = 0.5, 95% confidence interval (CI): 0.4, 0.8). A stronger relation was found among current smokers (OR = 0.3, 95% CI: 0.1, 0.7) than among ex-smokers (OR = 0.6, 95% CI: 0.4, 0.9), and there was an inverse gradient with pack-years smoked (trend p < 0.001). No associations were detected for coffee consumption or total caffeine intake or for alcohol consumption. However, reduced risks were observed for consumption of 2 cups/day or more of tea (OR = 0.4, 95% CI: 0.2, 0.9) and two or more cola drinks/day (OR = 0.6, 95% CI: 0.3, 1.4). The associations for tea and cola drinks were not confounded by smoking or coffee consumption. PMID- 11943692 TI - Effect of a school-based sun-protection intervention on the development of melanocytic nevi in children. AB - "Kidskin" was a 5-year (1995-1999), school-based intervention trial among first grade children in Perth, Western Australia. It aimed to assess whether a sun protection intervention could protect against nevus development on the trunk, face, and arms. Included were a control group, a "moderate intervention" group, and a "high intervention" group. Control schools taught the standard health curriculum, while intervention schools received a specially designed sun protection curriculum over 4 years. The high intervention group also received program materials over summer vacations when sun exposure was likely to be highest and were offered low-cost sun-protective swimwear. After adjustment for baseline nevus counts and potential confounding, nevus counts on all body sites were slightly lower in both intervention groups relative to the control group at follow-up, although the differences were not statistically significant and the high intervention was no more protective. Children in the moderate and high intervention groups, respectively, had fewer nevi on the back (6%, 95% confidence interval (CI): 0, 12; 4%, 95% CI: -3, 11), chest (boys) (5%, 95% CI: -4, 13; 3%, 95% CI: -8, 14), face (11%, 95% CI: 0, 21; 9%, 95% CI: -6, 21), and arms (8%, 95% CI: -1, 17; 3%, 95% CI: -10, 14). PMID- 11943693 TI - Dietary soy isoflavones and bone mineral density: results from the study of women's health across the nation. AB - Isoflavones are naturally occurring selective estrogen receptor modulators, with potential bone protective effects. To study the relation between soy isoflavone intake and bone mineral density (BMD), the authors analyzed baseline data from the Study of Women's Health Across the Nation, a US community-based cohort study of women aged 42-52 years. Their 1996-1997 analysis included African-American (n = 497), Caucasian (n = 1,003), Chinese (n = 200), and Japanese (n = 227) participants. Genistein and daidzein intakes were highly correlated (r = 0.98); therefore, analyses were conducted by using genistein. Median intakes of genistein (measured in micrograms/day) by African Americans and Caucasians were too low to pursue relational analyses further. For Chinese and Japanese women, median genistein intakes were 3,511 and 7,151 microg/day, respectively. Ethnic specific, linear models were used to predict BMD as a function of energy-adjusted tertile of intake, controlled for relevant covariates. For Chinese women, no association between genistein and BMD was found. Premenopausal, but not perimenopausal, Japanese women whose intakes were greater had higher spine and femoral neck BMD. Adjusted mean spinal BMD of those in the highest tertile of intake was 7.7% greater than that of women in the lowest tertile (p = 0.02); femoral neck BMD was 12% greater in the highest versus the lowest tertile (p < 0.0001). PMID- 11943694 TI - Validity of self-rated health among Latino(a)s. AB - The authors investigated whether self-rated health (SRH) had differential mortality risks for Latino(a) adults of various acculturation statuses living in the United States. They used cumulative National Health Interview Survey data from 1989 to 1994 (n = 37,713) linked with the National Health Interview Survey Multiple Cause of Death data files (1,364 deaths) that match records from the National Death Index through 1997. The authors specified survival models to estimate the effect of SRH on mortality and further stratified their model by birth and duration in the United States as proxies for acculturation. These estimates were compared across strata. Poor SRH was found to be a weaker predictor of subsequent mortality risk among the less acculturated, although the overall risk among the aggregated sample is similar to the risk reported in previous studies. The relation between poor SRH and mortality risk increases with United States acculturation among Latinos. While poor SRH was significantly associated with short-term mortality among the least acculturated, this association did not persist beyond 2-year mortality risk. Health researchers wishing to use SRH to assess the physical health of multiethnic populations should at least control for levels of acculturation among respondents. PMID- 11943696 TI - Commentary: Trade-offs in the development of a sample design for case-control studies. AB - The recent article, "Comparison of Telephone Sampling and Area Sampling: Response Rates and Within-Household Coverage" (Am J Epidemiol 2001;153:1119-27), raised a number of issues related to two sampling methodologies that can be used for selecting population-based controls for case-control studies: random digit dialing (RDD) and area probability sampling. Some of these issues are discussed in this commentary in more detail to help in making sample design decisions, including the need to take the analysis plan into account when developing a sample design. Data from the paper are used to illustrate how the choice of sample design can affect analyses. Relative costs associated with the two methodologies as well as variance and bias concerns are also discussed in detail. Sample coverage issues, including those associated with list-assisted RDD, are considered, as are some advantages of the list-assisted approach. A discussion of the use of concurrent screening and sampling with an RDD approach as an alternative to periodically selecting fixed sample sizes is provided. PMID- 11943695 TI - Evaluation of the effectiveness of highly active antiretroviral therapy in persons with human immunodeficiency virus using biomarker-based equivalence of disease progression. AB - The association of different CD4(+) cell counts with the same disease risk in treated and untreated populations reflects the effectiveness of highly active antiretroviral therapy (HAART) in persons with human immunodeficiency virus (HIV). Clinical progression of disease following initiation of HAART was determined for 679 HIV-infected men in the Multicenter AIDS Cohort Study by means of Kaplan-Meier survival analyses. Cox proportional hazards models were used to assess the effects of markers of HIV disease, antiretroviral history, and demographic factors. Men who had been followed since January 1993 (pre-HAART) were used to identify CD4(+) levels associated with the acquired immunodeficiency syndrome (AIDS)-free time equivalent to that of men starting HAART with CD4(+) cell counts of <200 cells/microl. Within 3.5 years following HAART initiation, 11.3% of the subjects developed AIDS and 8.5% died. Determinants of AIDS were a CD4(+) cell count of <200 cells/microl at initiation (relative hazard = 2.25, 95% confidence interval: 1.13, 4.49) and age >45 years at initiation (relative hazard = 1.92, 95% confidence interval: 0.98, 3.77). An increase in CD4(+) cell count of >50 cells/microl immediately after HAART initiation also improved prognosis (relative hazard = 0.34, 95% confidence interval: 0.16, 0.71). AIDS risk in men starting HAART with CD4(+) counts of <200 cells/microl (median = 132) was similar to that of non-HAART users with CD4(+) counts of 375-475 cells/microl (median = 432). The equivalence of disease progression to that of nonusers with approximately 300 more cells per microl demonstrates that HAART users have a broader reconstitution of the immune system beyond that of observed increases in CD4(+) cell count. PMID- 11943697 TI - Re: "Prevalence and incidence of hepatitis C virus infection in the US military: a sero-epidemiologic survey of 21,000 troops". PMID- 11943698 TI - Re: "Coffee consumption and serum lipids: a meta-analysis of randomized controlled clinical trials". PMID- 11943699 TI - Re: "Case-cohort analysis of agricultural pesticide applications near maternal residence and selected causes of fetal death". PMID- 11943703 TI - Alveolar soft part sarcoma: are we at the end or just the beginning of our quest? PMID- 11943704 TI - Clonal human (hNT) neuron grafts for stroke therapy: neuropathology in a patient 27 months after implantation. AB - Although grafted cells may be promising therapy for stroke, survival of implanted neural cells in the brains of stroke patients has never been documented. Human NT2N (hNT) neurons derived from the NTera2 (NT2) teratocarcinoma cell line were shown to remain postmitotic, retain a neuronal phenotype, survive >1 year in host rodent brains and ameliorate motor and cognitive impairments in animal models of ischemic stroke. Here we report the first postmortem brain findings of a phase I clinical stroke trial patient implanted with human hNT neurons adjacent to a lacunar infarct 27 months after surgery. Neurofilament immunoreactive neurons were identified in the graft site, fluorescent in situ hybridization revealed polyploidy in groups of cells at this site just like polyploid hNT neurons in vitro, and there was no evidence of a neoplasm. These findings indicate that implanted hNT neurons survive for >2 years in the human brain without deleterious effects. PMID- 11943705 TI - Heterogeneous methylation and deletion patterns of the INK4a/ARF locus within prostate carcinomas. AB - To elucidate the role of p53/p16(INK4a)/RB1 pathways in prostate carcinogenesis, we analyzed the p14(ARF), p16(INK4a), RB1, p21(Waf1), p27(Kip1), PTEN, p73, p53, and MDM2 gene status of multiple areas within 16 histologically heterogeneous prostate carcinomas using methylation-specific polymerase chain reaction, differential polymerase chain reaction, and immunohistochemistry. All focal areas examined had Gleason scores ranging from 1 to 5. Methylation of either PTEN or p73 was undetected in any sample, whereas expression of MDM2 seemed to be an independent event within small foci of 4 of 16 tumors. Loss of p14(ARF), p16(INK4a), RB1, and p27(Kip1) expression correlated with homozygous deletion or promoter hypermethylation. One carcinoma showed co-deletion of both p14(ARF) and p16(INK4a) in two of five areas examined; two areas within another tumor demonstrated concurrent hypermethylation of the promoter regions of the same genes. Focal hypermethylation of RB1, p21(Waf1), and p27(Kip1) was detected within two, two, and three tumors, respectively. These findings indicate that both genetic and epigenetic events occur independently in intratumor foci and further suggest hypermethylation-induced loss of gene function may be as critical as specific genetic mutations in prostate carcinogenesis. PMID- 11943707 TI - Diverse tumorigenic pathways in ovarian serous carcinoma. AB - This study was undertaken to analyze genetic alterations in 108 sporadic serous ovarian neoplasms to elucidate ovarian serous carcinogenesis. Our results demonstrate that K-ras mutations occur in approximately 50% of serous borderline tumors (SBTs), non-invasive micropapillary serous carcinomas (MPSCs), and invasive micropapillary serous carcinomas, which represent a morphological continuum of tumor progression. Moreover, progressive increase in the degree of allelic imbalance of chromosomes 1p, 5q, 8p, 18q, 22q, and Xp was observed comparing serous borderline tumors to noninvasive and invasive micropapillary serous carcinomas. In contrast, high-grade (conventional serous carcinoma) tumors contained wild-type K-ras in all 23 cases studied and a high frequency of allelic imbalance even in small (early) primary tumors similar to that found in advanced stage tumors. Based on these findings, we propose a dualistic model for ovarian serous carcinogenesis. One pathway involves a stepwise progression from SBT to noninvasive and then invasive MPSC. The other pathway is characterized by rapid progression from the ovarian surface epithelium or inclusion cysts to a conventional (high-grade) serous carcinoma. PMID- 11943706 TI - The precrystalline cytoplasmic granules of alveolar soft part sarcoma contain monocarboxylate transporter 1 and CD147. AB - Alveolar soft part sarcoma (ASPS) is an unusual tumor of young adults with the characteristic presence on ultrastructural analysis of rhomboid or rectangular cytoplasmic crystals. These membrane-bound crystals are known to form within specific PAS-diastase-resistant electron-dense cytoplasmic granules. The composition of these crystals and the dense granules from which they are derived has remained elusive. After the detection of strong discrete granular cytoplasmic immunoreactivity in ASPS for monocarboxylate transporter 1 (MCT1) in the course of a broad immunohistochemical characterization of an MCT1 antibody, we studied the expression of MCT1 and its interacting partner, CD147, in a panel of 10 ASPS cases using appropriate antibodies. MCT1 is one of a family of widely expressed proton-linked transporters for monocarboxylates such as lactate and pyruvate. In all normal and neoplastic tissues studied to date, MCT1 immunoreactivity is limited to the cell surface. We find that the periodic acid-Schiff-diastase resistant cytoplasmic granules of ASPS are strongly immunoreactive for MCT1 and CD147. Specifically, intense cytoplasmic granular positivity for MCT1 and CD147 was found in 7 of 10 and 8 of 10 ASPSs, respectively. Ultrastructural immunohistochemistry with immunogold labeling confirmed that the MCT1 immunoreactivity localized to the cytoplasmic electron-dense granules in ASPS. Western blot analysis of several ASPS cases confirmed that the protein reactive with the MCT1 antibody and that reactive with the CD147 antibody both migrated at the size expected for MCT1 and CD147, respectively. Thus, ASPS cells seem to accumulate MCT1-CD147 complexes in the specific cytoplasmic granules known to undergo crystallization. The possible basis for the overproduction or impaired surface localization of these proteins in ASPS remains unclear. PMID- 11943708 TI - Role of beta-catenin/T-cell factor-regulated genes in ovarian endometrioid adenocarcinomas. AB - In various cancers, inactivating mutations in the adenomatous polyposis coli or Axin tumor suppressor proteins or activating mutations in beta-catenin's amino terminal domain elevate beta-catenin levels, resulting in marked effects on T cell factor (TCF)-regulated transcription. Several candidate beta-catenin/TCF regulated genes in cancer have been proposed. Expression of a few of these genes has been studied in primary human cancers, but most studies have focused on colon cancers and not on other cancer types that harbor mutational defects in adenomatous polyposis coli, AXIN, or beta-catenin. Mutations leading to beta catenin deregulation are found in nearly half of ovarian endometrioid adenocarcinomas (OEAs). We report here on the expression of 6 candidate beta catenin/TCF-regulated genes in a panel of 44 primary OEAs, more than a third of which carry demonstrable defects in beta-catenin regulation. Using quantitative assays of gene expression, we found significantly elevated expression of the MMP 7, CCND1 (Cyclin D1), CX43 (Connexin 43), PPAR-delta, and ITF2 genes in OEAs with deregulated beta-catenin. This correlation was not observed for c-myc, another putative beta-catenin/TCF-regulated gene. Immunohistochemical studies confirmed that overexpression of cyclin D1 and MMP-7 was highly associated with nuclear accumulation of beta-catenin and mutational defects of the Wnt/beta-catenin/TCF signaling pathway. Our findings indicate cyclin D1, MMP-7, connexin 43, PPAR delta, and ITF-2, likely play important roles in the pathogenesis of those OEAs that manifest defects in beta-catenin regulation. PMID- 11943709 TI - Discovery of novel tumor markers of pancreatic cancer using global gene expression technology. AB - Despite several advances in our basic understanding and in the clinical management of pancreatic cancer, virtually all patients who will be diagnosed with pancreatic cancer will die from this disease. The high mortality of pancreatic cancer is predominantly because of diagnosis at an advanced stage of disease and a lack of effective treatments. We used the Gene Logic Inc. BioExpress platform and Affymetrix GeneChip arrays to identify genes differentially expressed in pancreatic cancer. cDNA was prepared from samples of normal pancreas (n = 11), normal gastrointestinal mucosa (n = 22), resected pancreas cancer tissues (n = 14), and pancreas cancer cell lines (n = 8), and was hybridized to the complete Affymetrix Human Genome U95 GeneChip set (arrays U95 A, B, C, D, and E) for simultaneous analysis of 60,000 cDNA fragments, with 12,000 fragments covering full-length genes and 48,000 fragments covering expressed sequence tags (ESTs). Genes expressed at levels at least fivefold greater in the pancreatic cancers ascompared to normal tissues were identified. Serial analysis of gene expression (SAGE) libraries (http://www.ncbi.nlm.nih.gov/SAGE/) of two normal pancreatic ductal cell cultures (HX and H126) were used to exclude genes expressed in the normal ducts (more than five tags per library). Differential expression of selected candidate genes was validated by immunohistochemical analysis (n = 3), by in situ hybridization (n = 1), and by reverse transcriptase-polymerase chain reaction (n = 8). One hundred eighty fragments were identified as having fivefold or greater expression levels in pancreas cancer specimens as compared to normal tissue, of which 124 corresponded to known genes and 56 to ESTs. Of these 124 fragments, 10 genes were represented by two or more fragments, resulting in 107 known genes identified as differentially expressed in pancreatic cancer. An additional 10 genes were expressed in the SAGE libraries of normal pancreatic duct epithelium, and were excluded from further analysis. A literature search indicated that 28 of the remaining 97 genes have been reported in association with pancreatic cancer, validating this approach. The remaining 69 genes have not been implicated in pancreatic cancer before, and have immediate potential as novel therapeutic targets and tumor markers of pancreatic cancer. PMID- 11943710 TI - Cervical epithelial cells transduced with the papillomavirus E6/E7 oncogenes maintain stable levels of oncoprotein expression but exhibit progressive, major increases in hTERT gene expression and telomerase activity. AB - Cervical carcinoma cells display high telomerase activity and usually contain and express integrated copies of the human papillomavirus (HPV) genome. Recent studies have demonstrated that the E6 oncogene of malignancy-associated HPVs increases cellular telomerase activity, predominantly via transcriptional activation of the catalytic subunit of telomerase, hTERT. To examine the relationship between E6 oncoprotein expression and telomerase expression during cellular immortalization, we transduced primary human cervical epithelial cells with the HPV E6/E7 genes and monitored temporal changes in viral oncoprotein expression, cellular hTERT RNA expression, and cellular telomerase activity. Quantitation of the individual E6 and E7 proteins, using a newly developed immunoprecipitation/immunoblotting technique, demonstrated that both oncoproteins were expressed at stable levels during successive passages of cervical cells. In contrast, the levels of hTERT mRNA and telomerase activity increased progressively and dramatically during passaging. Late-passage immortalized cells (passage 30) showed a 25-fold increase in hTERT mRNA and a 300-fold increase in telomerase activity compared to early-passage (passage 4) cells. Thus, neither hTERT mRNA expression nor telomerase activity are directly proportional to the level of E6 oncoprotein, indicating that E6 is not the sole determinant of the high levels of telomerase in cervical cells during immortalization. PMID- 11943712 TI - Levels of nonphosphorylated and phosphorylated tau in cerebrospinal fluid of Alzheimer's disease patients : an ultrasensitive bienzyme-substrate-recycle enzyme-linked immunosorbent assay. AB - We have developed an ultrasensitive bienzyme-substrate-recycle enzyme-linked immunosorbent assay for the measurement of Alzheimer's disease (AD) abnormally hyperphosphorylated tau in cerebrospinal fluid (CSF). The assay, which recognizes attomolar amounts of tau, is approximately 400 and approximately 1300 times more sensitive than conventional enzyme-linked immunosorbent assay in determining the hyperphosphorylated tau and total tau, respectively. With this method, we measured both total tau and tau phosphorylated at Ser-396/Ser-404 in lumbar CSFs from AD and control patients. We found that the total tau was 215 +/- 77 pg/ml in cognitively normal control (n = 56), 234 +/- 92 pg/ml in non-AD neurological (n = 37), 304 +/- 126 pg/ml in vascular dementia (n = 46), and 486 +/- 168 pg/ml (n = 52) in AD patients, respectively. However, a remarkably elevated level in phosphorylated tau was only found in AD (187 +/- 84 pg/ml), as compared with normal controls (54 +/- 33 pg/ml), non-AD (63 +/- 34 pg/ml), and vascular dementia (72 +/- 33 pg/ml) groups. If we used the ratio of hyperphosphorylated tau to total tau of > or =0.33 as cutoff for AD diagnosis, we could confirm the diagnosis in 96% of the clinically diagnosed patients with a specificity of 95%, 86%, 100%, and 94% against nonneurological, non-AD neurological, vascular dementia, and all of the three control groups combined, respectively. It is suggested that the CSF level of tau phosphorylated at Ser-396/Ser-404 is a promising diagnostic marker of AD. PMID- 11943711 TI - Telomere length assessment in human archival tissues: combined telomere fluorescence in situ hybridization and immunostaining. AB - A method was developed to assess human telomere lengths at the individual cell level in tissue sections from standard formalin-fixed paraffin-embedded tissues. We coupled this method with immunofluorescence to allow the simultaneous identification of specific cell types. Validation of this in situ quantification method showed excellent agreement with the commonly used telomere repeat fragment Southern blot method. The assay requires very few cells ( approximately 10 to 15). Thus, small tissue samples, including clinical biopsies, can be easily accommodated. In addition, the cells under study need not be actively cycling and there is no requirement for tissue disaggregation or cell culture. This method provides a more accurate assessment of telomere lengths than Southern blotting because confounding contributions from undesired cell types within tissue samples are avoided. Using this technique, we were able to perform the first comparison of relative telomere lengths in matched tumor versus normal epithelial cells within archival human prostate tissues. PMID- 11943714 TI - Expression of CD56/neural cell adhesion molecule correlates with the presence of lytic bone lesions in multiple myeloma and distinguishes myeloma from monoclonal gammopathy of undetermined significance and lymphomas with plasmacytoid differentiation. AB - Unlike monoclonal gammopathy of undetermined significance (MGUS) or non-Hodgkin's lymphomas (NHLs) with plasmacytoid differentiation, multiple myeloma (MM) is commonly associated with lytic bone lesions. Although the mechanisms of increased osteoclast activity are partially understood, comparatively little is known about the mechanisms that lead to the observed decrease in osteoblast function. Studies have shown neural cell adhesion molecule (NCAM) homophilic binding between MM cell lines and osteosarcoma cell lines, and that binding results in decreased osteoid production in vitro. Thus, we postulated that the expression of NCAM by MM cells contributes to lytic lesion formation by causing decreased osteoid production in vivo. We used immunohistochemistry in bone marrow core biopsies to assess NCAM expression in osteoblasts and plasma cells (PCs) in vitro. We found consistent, strong, uniform NCAM expression by the osteoblasts in all bone marrow core biopsies (352 of 352, 100%). Strong expression of NCAM by PCs correlated with the presence of lytic bone lesions (chi-square, 33.39: P <0.000; odds ratio, 16.9). There was also a strong correlation between NCAM expression and the diagnosis of MM in comparison to reactive PCs, MGUS, or NHLs with plasmacytoid differentiation (all P values <0.000). In conclusion, using immunohistochemistry, we found strong expression of NCAM by osteoblasts and that when equal to the intensity of osteoblast expression, NCAM expression by PCs correlates with the presence of lytic bone lesions and distinguishes MM from reactive plasmacytosis, NHLs with plasmacytoid differentiation, and most cases of MGUS. PMID- 11943713 TI - Neutralization of adrenomedullin inhibits the growth of human glioblastoma cell lines in vitro and suppresses tumor xenograft growth in vivo. AB - Presently, there is no effective treatment for glioblastoma, the most malignant and common brain tumor. Growth factors are potential targets for therapeutic strategies because they are essential for tumor growth and progression. Peptidylglycine alpha-amidating monooxygenase is the enzyme producing alpha amidated bioactive peptides from their inactive glycine-extended precursors. The high expression of peptidylglycine alpha-amidating monooxygenase mRNA in glioblastoma and glioma cell lines points to the involvement of alpha-amidated peptides in tumorigenic growth processes in the brain. After screening of amidated peptides, it was found that human glioblastoma cell lines express high levels of adrenomedullin (AM) mRNA, and that immunoreactive AM is released into the culture medium. AM is a multifunctional regulatory peptide with mitogenic and angiogenic capabilities among others. Real-time quantitative reverse transcriptase-polymerase chain reaction analysis showed that AM mRNA was correlated to the tumor type and grade, with high expression in all glioblastomas analyzed, whereas a low expression was found in anaplastic astrocytomas and barely detectable levels in low-grade astrocytomas and oligodendrogliomas. In the present study we also demonstrate the presence of mRNA encoding the putative AM receptors, calcitonin receptor-like receptor/receptor activity-modifying protein 2 and -3 (CRLR/RAMP2; CRLR/RAMP3) in both glioma tissues and glioblastoma cell lines and further show that exogenously added AM can stimulate the growth of these glioblastoma cells in vitro. These findings suggest that AM may function as an autocrine growth factor for glioblastoma cells. One way to test the autocrine hypothesis is to interrupt the function of the endogenously produced AM. Herein, we demonstrate that a polyclonal antibody specific to AM, blocks the binding of the hormone to its cellular receptors and decreases by 33% (P < 0.001) the growth of U87 glioblastoma cells in vitro. Intratumoral administration of the anti-AM antibody resulted in a 70% (P < 0.001) reduction in subcutaneous U87 xenograft weight 21 days after treatment. Furthermore, the density of vessels was decreased in the antibody-treated tumors. These findings support that AM may function as a potent autocrine/paracrine growth factor for human glioblastomas and demonstrate that inhibition of the action of AM (produced by tumor cells) may suppress tumor growth in vivo. PMID- 11943715 TI - Antibody response against perlecan and collagen types IV and VI in chronic renal allograft rejection in the rat. AB - Chronic rejection is the leading cause of late renal transplant failure. Various structural lesions are observed in grafts undergoing chronic rejection including glomerular basement membrane (GBM) duplications. The well-established Fisher (F344) to Lewis (LEW) rat renal transplant model for chronic rejection was used to assess the presence and role of the humoral immune response against graft antigens during chronic rejection. LEW recipients of F344 allografts develop transplant glomerulopathy and produce IgG1 antibodies directed against F344 GBM preparations that are detectable 3 weeks after transplantation. Glomerular IgG1 deposition was observed that in vitro co-localized with a rabbit anti-rat GBM antiserum in rejecting F344 grafts; elution experiments of isolated glomeruli yielded IgG1 antibodies reactive in vitro with F344 GBM, but not LEW GBM. Prevention of acute rejection by transient treatment of the recipients with cyclosporin A completely abrogated the production of anti-GBM antibodies. Using proteomic techniques we identified the antigens recognized by the LEW posttransplant sera as being the heparan sulfate proteoglycan perlecan and the alpha1 chain of collagen type VI in association with the alpha5 chain of collagen type IV. In conclusion, LEW recipients of F344 kidney grafts produce IgG1 antibodies against donor type perlecan and alpha1(VI)/alpha5(IV) collagen and develop transplant glomerulopathy. These data implicate an important role for the humoral immune response in the development of glomerulopathy during chronic rejection. PMID- 11943717 TI - Rapid activation of nuclear factor-kappaB in airway epithelium in a murine model of allergic airway inflammation. AB - Bronchiolar epithelium is postulated to play a critical role in the orchestration of responses to inhaled allergens, and may contribute to the pathogenesis of asthma. Using a murine model of allergic airway inflammation and hyperresponsiveness, we demonstrate in mice sensitized with ovalbumin (OVA) that following a single challenge with nebulized OVA, a rapid and protracted activation of inhibitor of kappa B kinase (IKK) occurred in lung tissue. IKK activation was followed by nuclear localization of nuclear factor (NF)-kappaB within the bronchiolar epithelium and increased luciferase activity in lungs of mice containing a NF-kappaB-dependent reporter gene. Challenge of sensitized mice with OVA also induced mRNA expression of the chemokines, macrophage inflammatory protein-2 (MIP-2) and eotaxin in lung tissue, which corresponded temporally with the observed influx of neutrophils and eosinophils, respectively, into the airspaces. Using laser capture microdissection and quantitative polymerase chain reaction, we demonstrated that MIP-2 and eotaxin were predominantly expressed in bronchiolar epithelium, in contrast to distal regions of the lungs, which expressed lower or undetectable levels of these mRNAs. These studies strengthen the potential importance of the bronchiolar epithelial cell as a source of production of NF-kappaB-dependent mediators that play a role in asthma. PMID- 11943716 TI - Human alpha-defensins HNPs-1, -2, and -3 in renal cell carcinoma: influences on tumor cell proliferation. AB - The alpha-defensins human neutrophil peptides (HNPs)-1, -2, and -3 have been described as cytotoxic peptides with restricted expression in neutrophils and in some lymphocytes. In this study we report that HNPs-1, -2, and -3 are also expressed in renal cell carcinomas (RCCs). Several RCC lines were found to express mRNA as well as the specific peptides of HNP-1, -2, and -3 demonstrated by reverse transcriptase-polymerase chain reaction, mass spectrometric, and flow cytometric analyses. At physiological concentrations HNPs-1, -2, and -3 stimulated cell proliferation of selected RCC lines in vitro but at high concentrations were cytotoxic for all RCC lines tested. As in RCC lines, alpha defensins were also detected in vivo in malignant epithelial cells of 31 RCC tissues in addition to their expected presence in neutrophils. In most RCC cases randomly, patchy immunostaining of alpha-defensins on epithelial cells surrounding neutrophils was seen, but in six tumors of higher grade malignancy all tumor cells were diffusely stained. Cellular necrosis observed in RCC tissues in association with extensive patches of HNP-1, -2, and -3, seemed to be related to high concentrations of alpha-defensins. The in vitro and in vivo findings suggest that alpha-defensins are frequent peptide constituents of malignant epithelial cells in RCC with a possible direct influence on tumor proliferation. PMID- 11943718 TI - Overexpression of endothelial nitric oxide synthase in endothelial cells is protective against ischemia-reperfusion injury in mouse skeletal muscle. AB - Microvascular injury has been proposed to be a main cause of ischemia-reperfusion (I/R) injury. The roles of endothelial nitric oxide synthase (eNOS)-derived NO, a key regulator of vascular function, in I/R injury are incompletely understood. We used transgenic mice overexpressing eNOS in endothelial cells (eNOS-Tg) and their littermates wild-type mice (WT) to investigate the roles of eNOS in I/R injury in skeletal muscle. Superoxide levels in the affected muscles were reduced by approximately 50% in eNOS-Tg compared with WT during reperfusion. In WT, the disassembly of endothelial junctional proteins seen in the early period of reperfusion was recovered in the later phase. These findings were correlated with the increased vascular permeability in vivo. In contrast, eNOS-Tg maintained the endothelial junction assembly as well as vascular permeability during reperfusion. Leukocyte extravasation into tissue and up-regulated expression of adhesion molecules in the reperfused vessels were significantly inhibited in eNOS Tg. Tissue viability of the affected muscle was decreased in WT time-dependently after reperfusion, whereas eNOS-Tg showed no significant reduction. NOS inhibition completely reversed these protective effects of eNOS overexpression in I/R injury. Thus, eNOS overexpression appears to prevent the I/R injury in skeletal muscle by maintaining vascular integrity. PMID- 11943719 TI - FEZ1/LZTS1 is down-regulated in high-grade bladder cancer, and its restoration suppresses tumorigenicity in transitional cell carcinoma cells. AB - FEZ1/LZTS1 is a tumor suppressor gene that maps to chromosome 8p22, a chromosomal region frequently deleted in many human malignancies, including transitional cell carcinoma (TCC) of the urinary bladder. FEZ1/LZTS1 alterations have been reported in esophageal, breast, prostate, and gastric carcinomas. Fez1 expression was studied in five TCC-derived cancer cell lines by Western blot analysis and in 60 primary TCCs of the urinary bladder by immunohistochemistry. Fez1 protein was absent or reduced in four of five cell lines and in 37 of 60 primary TCC examined. We also restored Fez1 protein expression in human SW780 TCC-derived cells lacking endogenous Fez1 protein to study the effects of Fez1 expression on cell proliferation, cell kinetics, and tumorigenicity in BALB/c nude mice. In vitro transduction of SW780 Fez1-negative cell, with Ad-FEZ1, inhibited cell growth, altered cell cycle progression, and suppressed subcutaneous tumor growth in nude mice. These results suggest that FEZ1/LZTS1 gene plays a role in the development of TCC of the urinary bladder by acting as a bona fide tumor suppressor gene both in vitro and in vivo. PMID- 11943720 TI - AMD3100, a CxCR4 antagonist, attenuates allergic lung inflammation and airway hyperreactivity. AB - The role of specific chemokine receptors during allergic asthmatic responses has been relatively undefined. A number of receptors are preferentially expressed on Th2 cells, including CCR4, CCR8, and CxCR4. In the present study, we have examined the role of CxCR4 in the development of cockroach allergen-induced inflammation and airway hyperreactivity in a mouse model of asthma. Using a specific inhibitor of CxCR4, AMD3100, our results indicate that blocking this receptor has a significant effect in down-regulating the inflammation and pathophysiology of the allergen-induced response. Treatment of allergic mice with AMD3100 significantly reduced airway hyperreactivity, peribronchial eosinophilia, and the overall inflammatory responses. In addition, there was a shift in the cytokine profile that was observed in the AMD3100-treated animals. Specifically, there was a significant reduction in interleukin-4 and interleukin-5 levels and a significant increase in interleukin-12 and interferon-gamma levels within the lungs of treated allergic mice. Furthermore, there was a significant alteration in the local chemokine production of CCL22 (MDC) and CCL17 (TARC), two chemokines previously shown to be important in Th2-type allergen responses. Overall, specifically blocking CxCR4 using AMD3100 reduced a number of pathological parameters related to asthmatic-type inflammation. PMID- 11943721 TI - Solid-pseudopapillary tumors of the pancreas are genetically distinct from pancreatic ductal adenocarcinomas and almost always harbor beta-catenin mutations. AB - Solid-pseudopapillary tumors (SPTs) are unusual pancreatic neoplasms of low malignant potential that most frequently affect young women. Genetic events contributing to the development of SPTs are unknown. Whereas the more common ductal adenocarcinomas of the pancreas essentially never harbor beta-catenin or APC gene mutations, we have recently identified alterations of the APC/beta catenin pathway in other nonductal pancreatic neoplasms including pancreatoblastomas and acinar cell carcinomas. We analyzed a series of 20 SPTs for somatic alterations of the APC/beta-catenin pathway using immunohistochemistry for beta-catenin protein accumulation, direct DNA sequencing of beta-catenin exon 3, and direct DNA sequencing of the mutation cluster region in exon 15 of the APC gene in those SPTs that did not harbor beta-catenin mutations. Immunohistochemical labeling for cyclin D1 was performed to evaluate the overexpression of this cell-cycle protein as one of the putative downstream effectors of beta-catenin dysregulation. In addition, we analyzed the SPTs for genetic alterations commonly found in pancreatic ductal adenocarcinomas, including mutations in the K-ras oncogene and p53 and DPC4 tumor suppressor genes, using direct DNA sequencing of K-ras and immunostaining for p53 and Dpc4. Almost all SPTs harbored alterations in the APC/beta-catenin pathway. Nuclear accumulation of beta-catenin protein was present in 95% (19 of 20), and activating beta-catenin oncogene mutations were identified in 90% (18 of 20) of the SPTs. Seventy-four percent (14 of 19) showed overexpression of cyclin D1, ranging from 10 to 70% of tumor nuclei. In contrast, no K-ras mutations were present in any of the 20 SPTs, and Dpc4 expression was intact in all 16 SPTs for which immunohistochemical labeling was successful. Overexpression of p53 was limited to only 3 of 19 (15.8%) SPTs. These results emphasize the two distinct, divergent genetic pathways of neoplastic progression in pancreatic ductal and nonductal neoplasms. PMID- 11943722 TI - A short mutational hot spot in the first intron of BCL-6 is associated with increased BCL-6 expression and with longer overall survival in large B-cell lymphomas. AB - BCL-6 somatic mutations have been described in normal and tumoral B lymphocytes, associated with germinal center transit. We analyzed mutations in the major mutation cluster of BCL-6 in a series of 45 large B-cell lymphomas (LBCLs) and 15 Burkitt's lymphomas, and their relation to the level of BCL-6 expression and clinical outcome. Mutations in LBCL cases revealed the existence of two distinct, short mutational hot spots, spanning positions 106 to 127 and 423 to 443, in which the mutation frequency was higher than expected (P < 0.001). Mutations in the 423 to 443 subcluster were associated with an increased level of expression, although this was not the case with the 106 to 127 cluster. Additionally, LBCL cases characterized by the presence of mutations in the 423 to 443 cluster showed an increased overall survival (P < 0.05) when compared with the nonmutated LBCL cases in these positions. Burkitt's lymphoma cases showed a slightly lower frequency of mutations with a nonclustered distribution and lacked any relationship with the level of expression or any clinical characteristic. Findings from LBCLs suggest that the 423 to 443 cluster includes a regulatory region that is of importance for BCL-6 expression. Deregulation of BCL-6 expression caused by these mutations could play an important role in lymphoma genesis or progression. PMID- 11943723 TI - Activation of the tie2 receptor by angiopoietin-1 enhances tumor vessel maturation and impairs squamous cell carcinoma growth. AB - The distinct roles of angiopoietin (Ang)-1 and Ang2, counteracting ligands for the endothelium-specific Tie2 receptor, in tumor development and progression have remained poorly understood. We investigated the expression of Ang1 and Ang2 during multistep mouse skin carcinogenesis and in human squamous cell carcinoma (SCC) xenografts. Expression of Ang2, but not of Ang1, was up-regulated in angiogenic tumor vessels already in early stages of skin carcinogenesis and was also strongly increased in SCCs. Stable overexpression of Ang1 in human A431 SCCs resulted in a more than 70% inhibition of tumor growth, associated with enhanced Tie2 phosphorylation levels, as compared with low levels in control transfected tumors. No major changes in the vascular density, vascular endothelial growth factor mRNA and protein expression, and vascular endothelial growth factor receptor-2 phosphorylation levels were observed in Ang1-expressing tumors. However, the fraction of tumor blood vessels with coverage by alpha-smooth muscle actin-positive periendothelial cells was significantly increased, indicative of an increased vascular maturation status. These findings identify an inhibitory role of Ang1/Tie2 receptor-mediated vessel maturation in SCC growth and suggest that up-regulation of its antagonist, Ang2, during early-stage epithelial tumorigenesis contributes to the angiogenic switch by counteracting specific vessel-stabilizing effects of Ang1. PMID- 11943724 TI - Expression of vascular endothelial growth factor and vascular endothelial growth factor receptor-2 (KDR/Flk-1) in ischemic skeletal muscle and its regeneration. AB - Vascular endothelial growth factor (VEGF) is a hypoxia-inducible endothelial cell mitogen and survival factor. Its receptor VEGFR-2 (KDR/Flk-1) mediates these effects. We studied the expression of VEGF and VEGFR-2 in ischemic human and rabbit skeletal muscle by immunohistochemistry and in situ hybridization. Human samples were obtained from eight lower limb amputations because of acute or chronic critical ischemia. In chronically ischemic human skeletal muscle VEGF and VEGFR-2 expression was restricted to atrophic and regenerating skeletal myocytes, whereas in acutely ischemic limbs VEGF and VEGFR-2 were expressed diffusely in the affected muscle. Hypoxia-inducible factor-1alpha was associated with VEGF and VEGFR-2 expression both in acute and chronic ischemia but not in regeneration. Hindlimb ischemia was induced in 20 New Zealand White rabbits by excising the femoral artery. Magnetic resonance imaging and histological sections revealed extensive ischemic damage in the thigh and leg muscles of ischemic rabbit hindlimbs with VEGF expression similar to acute human lower limb ischemia. After 1 and 3 weeks of ischemia VEGF expression was restricted to regenerating myotubes and by 6 weeks regeneration and expression of VEGF was diminished. VEGFR-2 expression was co-localized with VEGF expression in regenerating myotubes. Macrophages and an increased number of capillaries were associated with areas of ischemic muscle expressing VEGF and VEGFR-2. In conclusion, two patterns of VEGF and VEGFR-2 expression in human and rabbit ischemic skeletal muscle are demonstrated. In acute skeletal muscle ischemia VEGF and VEGFR-2 are expressed diffusely in the affected muscle. In chronic skeletal muscle ischemia and in skeletal muscle recovering from ischemia VEGF and VEGFR-2 expression are restricted to atrophic and regenerating muscle cells suggesting the operation of an autocrine pathway that may promote survival and regeneration of myocytes. PMID- 11943726 TI - Evidence for an active inflammatory process in the hibernating human myocardium. AB - Myocardial hibernation refers to a state of prolonged impairment of left ventricular function in the presence of coronary artery disease, which may be reversed by revascularization. In this study we present evidence for a local inflammatory reaction in hibernating myocardial segments from patients undergoing coronary revascularization. We obtained transmural myocardial biopsies guided by transesophageal echocardiography from patients with ischemic ventricular dysfunction undergoing bypass surgery. Among the 28 biopsied segments included in the study, 23 showed evidence of systolic dysfunction. The majority of dysfunctional segments (85.7%) were viable ((201)Tl uptake >/= 60%). The samples were stained with markers for mast cells, mature resident macrophages, and the monoclonal antibody Mac387 that labels newly recruited myeloid cells. Dysfunctional segments showed more extensive fibrosis and higher macrophage density than normal segments. Among the 23 dysfunctional segments, 12 recovered function as assessed with echocardiograms 3 months after revascularization. Segments with postoperative functional recovery had comparable macrophage and mast cell density with those showing persistent dysfunction. However, biopsied segments that subsequently recovered function contained significantly higher numbers of newly recruited Mac387-positive leukocytes (18.7 +/- 3.1 cells/mm(2), n = 12 versus 8.6 +/- 0.9 cells/mm(2), n = 11; P = 0.009). In addition, monocyte chemotactic protein-1, a potent mononuclear cell chemoattractant, was predominantly expressed in segments with recovery of function. Myocardial hibernation is associated with an inflammatory response leading to active leukocyte recruitment. Dysfunctional myocardial segments that show an active inflammatory reaction have a greater potential for recovery of function after revascularization. We postulate that revascularization may promote resolution of the ongoing inflammation, preventing further tissue injury and fibrosis. PMID- 11943725 TI - Vascular endothelial growth factor ligands and receptors that regulate human cytotrophoblast survival are dysregulated in severe preeclampsia and hemolysis, elevated liver enzymes, and low platelets syndrome. AB - Human placental development combines elements of tumorigenesis and vasculogenesis. The organ's specialized epithelial cells, termed cytotrophoblasts, invade the uterus where they reside in the interstitial compartment. They also line uterine arteries and veins. During invasion, ectodermally derived cytotrophoblasts undergo pseudovasculogenesis, switching their adhesion molecule repertoire to mimic that of vascular cells. Failures in this transformation accompany the pregnancy complication preeclampsia. Here, we used a combination of in situ and in vitro analyses to characterize the cell's expression of vascular endothelial growth factor (VEGF) family ligands and receptors, key regulators of conventional vasculogenesis and angiogenesis. Cytotrophoblast differentiation and invasion during the first and second trimesters of pregnancy were associated with down-regulation of VEGF receptor (VEGFR)-2. Invasive cytotrophoblasts in early gestation expressed VEGF-A, VEGF-C, placental growth factor (PlGF), VEGFR-1, and VEGFR-3 and, at term, VEGF-A, PlGF, and VEGFR-1. In vitro the cells incorporated VEGF-A into the surrounding extracellular matrix; PlGF was secreted. We also found that cytotrophoblasts responded to the VEGF ligands they produced. Blocking ligand binding significantly decreased their expression of integrin alpha1, an adhesion molecule highly expressed by endovascular cytotrophoblasts, and increased apoptosis. In severe preeclampsia and hemolysis, elevated liver enzymes, and low platelets syndrome, immunolocalization on tissue sections showed that cytotrophoblast VEGF A and VEGFR-1 staining decreased; staining for PlGF was unaffected. Cytotrophoblast secretion of the soluble form of VEGFR-1 in vitro also increased. Together, the results of this study showed that VEGF family members regulate cytotrophoblast survival and that expression of a subset of family members is dysregulated in severe forms of preeclampsia. PMID- 11943727 TI - Primary thymic extranodal marginal-zone B-cell lymphoma of mucosa-associated lymphoid tissue type exhibits distinctive clinicopathological and molecular features. AB - Extranodal marginal-zone B-cell lymphoma (MZBL) of mucosa-associated lymphoid tissue (MALT) arising in the thymus is rare, with the largest series in the literature including only three cases. In the present study, we investigated 15 cases of thymic MALT lymphoma to systematically characterize its clinical, histopathological, and molecular features. There was a marked female predilection (male:female = 1:4), with a mean age of 55 years at diagnosis. There was a strong association with autoimmune disease, especially Sjogren's syndrome. Histologically, the thymic lymphoma showed the characteristic morphological features of extranodal MZBL of MALT type. Cysts were common. Prominent lymphoepithelial lesions were formed by centrocyte-like cells infiltrating and expanding the Hassall's corpuscles and epithelium lining the cysts. Plasmacytic differentiation was apparent in all cases. Notably, 13 of 15 cases expressed immunoglobulin (Ig) A phenotype; IgA expression in thymic MALT lymphoma was in striking contrast with the IgM phenotype observed in most of the Sjogren's syndrome-associated MZBLs and MALT lymphomas at other sites. Epstein-Barr virus was absent, and API2-MALT1 gene fusion, a recently reported MALT lymphoma specific gene abnormality, was not detected in any case. Although one patient died of disease 85 months after the diagnosis, other patients were alive with overall 3-year and 5-year survival rates being 89% and 83%, respectively. Among the 22 patients reported previously and in the present series, at least 17 patients (77%) were Asians. These data indicate that thymic MALT lymphoma may represent a distinct subgroup of MALT lymphoma characterized by apparent predilection for Asians, a strong association with autoimmune disease, frequent presence of cysts, consistent plasma cell differentiation, tumor cells expressing IgA phenotype, and consistent lack of API2-MALT1 gene fusion. PMID- 11943728 TI - Hepatic expression of secondary lymphoid chemokine (CCL21) promotes the development of portal-associated lymphoid tissue in chronic inflammatory liver disease. AB - The chronic inflammatory liver disease primary sclerosing cholangitis (PSC) is associated with portal inflammation and the development of neolymphoid tissue in the liver. More than 70% of patients with PSC have a history of inflammatory bowel disease and we have previously reported that mucosal addressin cell adhesion molecule-1 is induced on dendritic cells and portal vascular endothelium in PSC. We now show that the lymph node-associated chemokine, CCL21 or secondary lymphoid chemokine, is also strongly up-regulated on CD34(+) vascular endothelium in portal associated lymphoid tissue in PSC. In contrast, CCL21 is absent from LYVE-1(+) lymphatic vessel endothelium. Intrahepatic lymphocytes in PSC include a population of CCR7(+) T cells only half of which express CD45RA and which respond to CCL21 in migration assays. The expression of CCL21 in association with mucosal addressin cell adhesion molecule-1 in portal tracts in PSC may promote the recruitment and retention of CCR7(+) mucosal lymphocytes leading to the establishment of chronic portal inflammation and the expanded portal-associated lymphoid tissue. This study provides further evidence for the existence of portal associated lymphoid tissue and is the first evidence that ectopic CCL21 is associated with lymphoid neogenesis in human inflammatory disease. PMID- 11943730 TI - Expression of UGT2B7, a UDP-glucuronosyltransferase implicated in the metabolism of 4-hydroxyestrone and all-trans retinoic acid, in normal human breast parenchyma and in invasive and in situ breast cancers. AB - Glucuronidation, mediated by UDP-glucuronosyltransferases (UGTs), affects the actions and disposition of diverse endo- and xenobiotics. In the case of catecholestrogens (CEs), glucuronidation is likely to block their oxidation to quinone estrogens that are the putative mediators of CEs' actions as initiators of cancers. The goal of this study was to determine whether UGT2B7, the isoenzyme with a high affinity for 4-hydroxyestrone, is expressed in human breast parenchyma. Glucuronidation of 4-hydroxyestrone has relevance to breast carcinogenesis because quinone metabolites of 4-hydroxylated CEs can form potentially mutagenic depurinating DNA adducts, and because in breast tissue estrone is likely to be the predominant estrogen available for 4-hydroxylation. Using reverse transcriptase-polymerase chain reaction, immunocytochemistry, immunoblot analyses, and assays of glucuronidation of 4-hydroxyestrone, we show that UGT2B7 is expressed in human mammary epithelium, and that its expression is dramatically reduced in invasive breast cancers. In many in situ carcinomas, however, 4-hydroxyestrone immunostaining was not only preserved but even more intense than in normal mammary epithelium. The finding of reduced UGT2B7 protein and glucuronidation of 4-hydroxyestrone in invasive cancers suggests a tumor suppressor function for the enzyme. Recent identification of all-trans retinoic acid as a substrate of UGT2B7 suggests that this function includes the generation of retinoyl-beta-glucuronide, a potent mediator of actions of retinoids important for maintaining epithelia in a differentiated state. Current knowledge does not provide any ready explanation for the apparent increase in UGT2B7 expression in carcinomas in situ. However, this finding, together with reduced immunostaining at loci showing breach of the basement membrane (microinvasion), suggests involvement of UGT2B7-catalyzed reaction(s) in protection against invasion of surrounding tissue by cancer cells. PMID- 11943729 TI - S-phase kinase-associated protein 2 expression in non-Hodgkin's lymphoma inversely correlates with p27 expression and defines cells in S phase. AB - The protein expression of the cyclin-dependent kinase inhibitor p27 is often deregulated in human tumors. In lymphomas the inactivation of p27 is achieved through either increased degradation(1) or sequestration via D cyclins,(2) and p27 protein levels have been shown to have a prognostic significance.(1,3) Recently, S-phase kinase-associated protein 2 (Skp2) has been proved to mediate p27 degradation in normal cells(4-7) and to have oncogenetic properties.(8,9) In this study, B-, T-, and myeloid hematopoietic cell lines and a well-characterized panel of human lymphomas (n = 244) were studied for the expression of Skp2. In human lymphomas, the expression of Skp2 strongly related to the grade of malignancy, being low in indolent tumors and very high in aggressive lymphomas. Moreover, the percentages of Skp2- and S-phase-positive cells, as measured by DNA content or BrdU labeling, strictly matched and closely parallel that of Ki-67 and cyclin A. An inverse correlation between Skp2 and p27 was found in the majority of lymphoma subtypes. Nonetheless, most mantle cell lymphomas and a subset of diffuse large cell lymphomas failed to show this correlation, suggesting that alternative pathway(s) for the regulation of p27 might exist. The detection of Skp2 protein either by flow cytometry or by immunohistochemistry represents a simple method to precisely assess the S phase of lymphomas. The potential diagnostic and prognostic value of Skp2 is discussed. PMID- 11943731 TI - DNA mismatch repair deficiency accelerates endometrial tumorigenesis in Pten heterozygous mice. AB - PTEN mutation and microsatellite instability are two of the most common genetic alterations in uterine endometrioid carcinoma. Furthermore, previous studies have suggested an association between the two alterations, however the basis and consequence of the association is not understood. Recently it has been shown that 100% of female Pten(+/-) mice develop complex atypical hyperplasia by 32 weeks of age that progresses to endometrial carcinoma in approximately 20 to 25% of mice at 40 weeks. In an attempt to expand this mouse model of endometrial tumorigenesis and to further our understanding of the association betweenPten mutations and DNA mismatch repair deficiency, we generated Ptenheterozygous, Mlh1 null (mismatch repair deficient) mice. Significantly, the majority ofPten(+/ )/Mlh1(-/-)mice developed polypoid lesions in the endometrium at 6 to 9 weeks of age. By 14 to 18 weeks, all of the double-mutant mice had lesions histologically similar to those seen inPten(+/-) mice, and two of them exhibited invasive disease. Moreover, the frequency of loss of the wild-type Pten allele in the double-mutant mice at 14 to 18 weeks was similar to that seen in lesions from 40 week-old Pten(+/-) mice. Taken together, our results indicate that DNA mismatch repair deficiency can accelerate endometrial tumorigenesis inPten heterozygous mice and suggests that loss of the wild-type Pten allele is involved in the development/progression of tumors in this setting. PMID- 11943733 TI - Induction of the hyaluronic acid-binding protein, tumor necrosis factor stimulated gene-6, in cervical smooth muscle cells by tumor necrosis factor-alpha and prostaglandin E(2). AB - Immediately before parturition the cervix undergoes striking changes in structure (ripening) that facilitate dilatation and effacement. Cervical ripening shares many features in common with inflammation-associated tissue remodeling, making it a valuable process to explore with respect to the biochemical events in extracellular matrix restructuring. Cervical ripening can be pharmacologically induced with prostaglandin E(2) (PGE(2)). Among the biochemical changes in the cervix at parturition is a marked increase in the hyaluronic acid (HA) content. HA and HA-binding proteins have been implicated in tissue hydration, release of collagenase, and leukocyte migration, but their roles in cervical ripening have not been explored. In the present study we examined the ability of PGE(2) to induce expression of the HA-binding protein, tumor necrosis factor-stimulated gene (TSG)-6, in human cervical smooth muscle cells (hCSMCs) and compared the PGE(2) response to that of tumor necrosis factor-alpha (TNF-alpha), an established inducer of TSG-6. TNF-alpha stimulated TSG-6 mRNA accumulation in a dose- and time-dependent manner, with the maximal response observed at 10 ng/ml after 6 hours of incubation. PGE(2) stimulated TSG-6 mRNA expression, but the magnitude of response was substantially less than that produced by TNF-alpha, and it was maximal only after 24 hours of incubation. Quantitative real-time polymerase chain reaction was performed to assess the induction of TSG-6 mRNA and nascent transcripts at 24 hours of treatment. Induction of TSG-6 mRNA and nascent transcripts in response to 10 micromol/L of PGE(2) was 5.7-fold and 6.3-fold greater than control values, respectively, whereas TNF-alpha (10 ng/ml) induced TSG-6 mRNA and nascent transcripts by 80-fold and 134-fold, respectively. TNF alpha and PGE(2) stimulated secretion of TSG-6 into the culture medium as detected by Western blotting. The effects of PGE(2) on secretion of TSG-6 were delayed compared to TNF-alpha. A 1.3-kb fragment of the human TSG-6 proximal promoter drove luciferase expression in transfected hCSMCs. PGE(2) increased TSG 6 promoter activity 1.75-fold. Paradoxically, TNF-alpha reduced TSG-6 promoter activity by 50%. We conclude that hCSMCs express the hyaladherin TSG-6; that TSG 6 expression in these cells is regulated by PGE(2) as well as proinflammatory cytokines; responses of hCSMCs to TNF-alpha and PGE(2) are distinct in terms of magnitude and the time course; and PGE(2) and TNF-alpha exert different effects on the TSG-6 proximal promoter. PMID- 11943732 TI - Diversity of genomic breakpoints in TFG-ALK translocations in anaplastic large cell lymphomas: identification of a new TFG-ALK(XL) chimeric gene with transforming activity. AB - Anaplastic large cell lymphomas are associated with chromosomal aberrations involving the anaplastic lymphoma kinase (ALK) gene at 2p23 that result in the expression of novel chimeric ALK proteins with transforming properties. In most of these tumors, the t(2;5)(p23;q35) generates the NPM-ALK fusion gene. However, several studies have now demonstrated that genes other than NPM may be fused to the ALK gene. We have recently described two different ALK rearrangements involving the TRK-fused gene (TFG) in which the same portion of ALK was fused to different length fragments of the 5' TFG region. These two rearrangements encoded chimeric proteins of 85 kd (TFG-ALK(S)) and 97 kd (TFG-ALK(L)), respectively. In this study, we have identified a new ALK rearrangement in which the catalytic domain of ALK was fused to a larger fragment of the TFG gene (TFG-ALK(XL)), encoding for a fusion protein of 113 kd. Genomic analysis of these three TFG-ALK rearrangements revealed that the TFG breakpoints occur at introns 3, 4, and 5, respectively, whereas the ALK breakpoints always occur in the same intron. No homologous regions or known recombination sequences were found in these regions. Transfection experiments using NIH-3T3 fibroblasts showed a similar transforming efficiency of TFG-ALK variants compared with NPM-ALK. In addition, in common with NPM-ALK, the TFG-ALK proteins formed stable complexes with the signaling proteins Grb2, Shc, and PLC-gamma. In conclusion, these findings indicate that the TFG may use a variety of intronic breakpoints in ALK rearrangements generating fusion proteins of different molecular weights, but with similar transforming potential than NPM-ALK. PMID- 11943734 TI - PolyA deletions in hereditary nonpolyposis colorectal cancer: mutations before a gatekeeper. AB - Microsatellite instability (MSI) secondary to loss of DNA mismatch repair (MMR) is present in adenomas and colorectal carcinomas from individuals with hereditary nonpolyposis colorectal cancer (HNPCC). To better characterize when MMR loss occurs during HNPCC progression, the extent of deletions in noncoding polyA sequences were compared between 6 adenomas (all < or = 1.0 cm in size) and 10 cancers. Numbers of deleted bases reflect time since loss of MMR because polyA deletions are stepwise. Adenoma deletions were nearly the same (85%) as the cancers with sum total deletions at four different polyA loci of -32.7 bases in adenomas and -38.4 bases in cancers. Intervals between negative clinical examinations and tumor removal (average of 2.1 years) were known for six tumors. There were no significant differences in the extent of deletions in tumors removed under clinical surveillance (-34.8 bases) versus tumors removed without prior negative examinations (-36.5 bases). These findings illustrate that MSI is extensive in both small adenomas, and tumors which appear after negative clinical examinations, consistent with an early loss of MMR in HNPCC, even before a gatekeeper mutation. PMID- 11943735 TI - Type I interferon gene transfer sensitizes melanoma cells to apoptosis via a target activity on mitochondrial function. AB - Our previous article reported that retroviral transduction of human type I consensus interferon-coding sequence into two human melanoma cells increased their susceptibility to cisplatin-induced apoptosis. Importantly, primary melanoma cells were significantly more sensitive to cisplatin-induced apoptosis with respect to metastatic melanoma cells. The aim of this study was to elucidate the subcellular mechanisms involved in this interferon-induced apoptotic proneness. Our results indicate that 1) cisplatin-induced apoptosis can be referred to as the type II apoptosis, ie, to the mitochondrially driven cascade; 2) treatment of interferon-producing melanoma cells with other type II apoptotic stimuli, such as radiation or staurosporine, also resulted in massive apoptosis, whereas type I stimuli, ie, anti-Fas, were ineffective; 3) interferon sensitization involved the caspase cascade in primary melanoma cells and the alternative pathway represented by cathepsin-mediated apoptosis in metastatic melanoma cells; 4) interferon production sensitizes cells to apoptosis by inducing, as the earliest event, mitochondrial membrane hyperpolarization. These results suggest that constitutive production of type I interferon by melanoma cells can act as an intracellular booster capable of increasing cell proneness to apoptosis by specifically modifying mitochondrial homeostasis and independently from the apoptotic cascade involved. PMID- 11943736 TI - Constitutive expression of c-FLIP in Hodgkin and Reed-Sternberg cells. AB - Crosslinking of the transmembrane receptor CD95/Fas leads to activation of a signaling cascade resulting in apoptosis. c-FLIP is a recently described protein that potently inhibits Fas-mediated apoptosis and has been shown to be a key factor in germinal center B cell survival. Because Hodgkin and Reed-Sternberg cells in classical Hodgkin's disease (cHD) are also resistant to Fas-mediated apoptosis we studied the role of c-FLIP in classical HD. High levels of c-FLIP protein were identified in two Fas-resistant Hodgkin-derived cell lines. In contrast to other tumor cells, inhibition of protein synthesis by cycloheximide did not lead to down-regulation of c-FLIP protein in these HD cell lines. Furthermore, Fas-mediated apoptosis was only partially restored suggesting that normal regulation of c-FLIP was disrupted. The in vivo relevance of these findings was supported by demonstration of significant c-FLIP expression by immunohistochemistry in 18 of 19 evaluable cases of primary HD. Taken together, c FLIP is constitutively expressed in HD and may therefore be a major mechanism responsible for Fas-resistance in HD. PMID- 11943738 TI - Beckwith-Wiedemann syndrome, pancreatoblastoma, and the wnt signaling pathway. PMID- 11943737 TI - Pathology of interstitial cells of Cajal in relation to inflammation revealed by ultrastructure but not immunohistochemistry. AB - The role of interstitial cells of Cajal associated with Auerbach's plexus (ICC AP) in the pathophysiology of inflammation-induced abnormalities in gut motor activity is poorly understood. Therefore we applied a well-described model of inflammation (infection by Trichinella spiralis) to the mouse small intestine where the structure and function of ICC-AP are best known. Electron microscopic evaluation revealed that 1 to 3 days after infection, selective and patchy damage to the ICC processes occurred, thereby disrupting contacts between these ICC and smooth muscle cells as well as ICC and nerves, which was associated with disordered electrical activity and abnormal peristalsis. Ten to 15 days after infection, damage to ICC-AP was maximal and now involving the cell body and major processes. Marked synthetic activity and regrowth of their processes occurred from day 3 onward and recovery was completed at day 40 after infection. No changes to the network of ICC-AP were seen with c-Kit immunohistochemistry. From day 1 after infection, macrophages infiltrated the AP area, making close contact including peg-and-socket-like junctions with smooth muscle cells and ICC-AP but up to day 6 after infection without any sign of phagocytosis. By day 6 after infection, lymphocytes entered the musculature forming close contacts with ICC AP. This was not associated with damage to ICC-AP but with proliferation of rough endoplasmic reticulum. From day 23 onward, immune cells withdrew from the musculature except macrophages, resulting in a markedly increased population of macrophages in the AP area at day 60 after infection. PMID- 11943739 TI - KAI1 metastasis suppressor protein in cervical cancer. PMID- 11943740 TI - Tissue factor identifies cardiomyocyte apoptotic bodies in reperfusion injury. PMID- 11943741 TI - The lutropin/choriogonadotropin receptor, a 2002 perspective. AB - Reproduction cannot take place without the proper functioning of the lutropin/choriogonadotropin receptor (LHR). When the LHR does not work properly, ovulation does not occur in females and Leydig cells do not develop normally in the male. Also, because the LHR is essential for sustaining the elevated levels of progesterone needed to maintain pregnancy during the first trimester, disruptions in the functions of the LHR during pregnancy have catastrophic consequences. As such, a full understanding of the biology of the LHR is essential to the survival of our species. In this review we summarize our current knowledge of the structure, functions, and regulation of this important receptor. PMID- 11943742 TI - Androgen receptor (AR) coregulators: an overview. AB - The biological action of androgens is mediated through the androgen receptor (AR). Androgen-bound AR functions as a transcription factor to regulate genes involved in an array of physiological processes, most notably male sexual differentiation and maturation, and the maintenance of spermatogenesis. The transcriptional activity of AR is affected by coregulators that influence a number of functional properties of AR, including ligand selectivity and DNA binding capacity. As the promoter of target genes, coregulators participate in DNA modification, either directly through modification of histones or indirectly by the recruitment of chromatin-modifying complexes, as well as functioning in the recruitment of the basal transcriptional machinery. Aberrant coregulator activity due to mutation or altered expression levels may be a contributing factor in the progression of diseases related to AR activity, such as prostate cancer. AR demonstrates distinct differences in its interaction with coregulators from other steroid receptors due to differences in the functional interaction between AR domains, possibly resulting in alterations in the dynamic interactions between coregulator complexes. PMID- 11943743 TI - Disordered fat storage and mobilization in the pathogenesis of insulin resistance and type 2 diabetes. AB - The primary genetic, environmental, and metabolic factors responsible for causing insulin resistance and pancreatic beta-cell failure and the precise sequence of events leading to the development of type 2 diabetes are not yet fully understood. Abnormalities of triglyceride storage and lipolysis in insulin sensitive tissues are an early manifestation of conditions characterized by insulin resistance and are detectable before the development of postprandial or fasting hyperglycemia. Increased free fatty acid (FFA) flux from adipose tissue to nonadipose tissue, resulting from abnormalities of fat metabolism, participates in and amplifies many of the fundamental metabolic derangements that are characteristic of the insulin resistance syndrome and type 2 diabetes. It is also likely to play an important role in the progression from normal glucose tolerance to fasting hyperglycemia and conversion to frank type 2 diabetes in insulin resistant individuals. Adverse metabolic consequences of increased FFA flux, to be discussed in this review, are extremely wide ranging and include, but are not limited to: 1) dyslipidemia and hepatic steatosis, 2) impaired glucose metabolism and insulin sensitivity in muscle and liver, 3) diminished insulin clearance, aggravating peripheral tissue hyperinsulinemia, and 4) impaired pancreatic beta-cell function. The precise biochemical mechanisms whereby fatty acids and cytosolic triglycerides exert their effects remain poorly understood. Recent studies, however, suggest that the sequence of events may be the following: in states of positive net energy balance, triglyceride accumulation in "fat-buffering" adipose tissue is limited by the development of adipose tissue insulin resistance. This results in diversion of energy substrates to nonadipose tissue, which in turn leads to a complex array of metabolic abnormalities characteristic of insulin-resistant states and type 2 diabetes. Recent evidence suggests that some of the biochemical mechanisms whereby glucose and fat exert adverse effects in insulin-sensitive and insulin-producing tissues are shared, thus implicating a diabetogenic role for energy excess as a whole. Although there is now evidence that weight loss through reduction of caloric intake and increase in physical activity can prevent the development of diabetes, it remains an open question as to whether specific modulation of fat metabolism will result in improvement in some or all of the above metabolic derangements or will prevent progression from insulin resistance syndrome to type 2 diabetes. PMID- 11943744 TI - Predictive value of hormone measurements in maternal and fetal complications of pregnancy. AB - Intrauterine tissues (placenta, amnion, chorion, decidua) express hormones and cytokines that play a decisive role in maternal-fetal physiological interactions. The excessive or deficient release of some placental hormones in association with gestational diseases may reflect an abnormal differentiation of the placenta, an impaired fetal metabolism, or an adaptive response of the feto-placental unit to adverse conditions. This review is focused on the applicability of hormone measurements in the risk assessment, early diagnosis, and management of pregnancies complicated by Down's syndrome, fetal growth restriction, preeclampsia, preterm delivery, and diabetes mellitus. Combined hormonal tests or the combination of hormones and ultrasound may achieve reasonable sensitivity, but research continues to simplify the screening programs without sacrificing their accuracy. Only in a few instances is there sufficient evidence to firmly recommend the routine use of hormone tests to predict maternal and fetal complications, but the judicious use of selected tests may enhance the sensitivity of the risk assessment based solely on clinical and ultrasound examination. PMID- 11943750 TI - Understanding life--now for the hard part! PMID- 11943747 TI - The epithelial Na+ channel: cell surface insertion and retrieval in Na+ homeostasis and hypertension. AB - The epithelial Na+ channel (ENaC) forms the pathway for Na+ absorption in the kidney collecting duct and other epithelia. Dominant gain-of-function mutations cause Liddle's syndrome, an inherited form of hypertension resulting from excessive renal Na+ absorption. Conversely, loss-of-function mutations cause pseudohypoaldosteronism type I, a disorder of salt wasting and hypotension. Thus, ENaC has a critical role in the maintenance of Na+ homeostasis and blood pressure control. Altered Na+ absorption in the lung may also contribute to the pathogenesis of cystic fibrosis. Epithelial Na+ absorption is regulated in large part by mechanisms that control the expression of ENaC at the cell surface. Nedd4, a ubiquitin protein ligase, binds to ENaC and targets the channel for endocytosis and degradation. Liddle's syndrome mutations disrupt the interaction between ENaC and Nedd4, resulting in an increase in the number of ENaC channels at the cell surface. Aldosterone and vasopressin also regulate Na+ absorption to defend against hypotension and hypovolemia. Both hormones increase the expression of ENaC at the cell surface. The goal of this review is to summarize recent data on the regulation of ENaC expression at the cell surface. PMID- 11943751 TI - Painful memories. Can we train chronic pain patients to 'forget' their pain? PMID- 11943752 TI - A European Research Council--more competition in science. An interview with Enric Banda, Secretary General of the European Science Foundation. PMID- 11943753 TI - Learning from PISA. Reasons and remedies for student under-performance in reading, maths and science. PMID- 11943754 TI - The cutting edge in surgery. Telesurgery has been shown to be feasible--now it has to be made economically viable. PMID- 11943755 TI - Tapping into the pool of women. Facing a serious shortage of skilled workers, Europe's high-tech economies are discovering the value of working women. PMID- 11943758 TI - Evolution in a nutshell. EMBL PhD student symposium on evolution. PMID- 11943759 TI - The winding road from adhesive receptors to the nucleus. Conference on molecular biology of cellular interactions: adhesion receptor signalling and regulation of gene expression. PMID- 11943760 TI - Those magnificent molecular machines: logistics in dsRNA virus transcription. PMID- 11943761 TI - Chromatin remodeling enzymes: taming the machines. Third in review series on chromatin dynamics. AB - Members of the ATP-dependent family of chromatin remodeling enzymes play key roles in the regulation of transcription, development, DNA repair and cell cycle. Each of these enzymes are multi-subunit assemblies that hydrolyze thousands of molecules of ATP in order to change nucleosome positions, disrupt DNA-histone interactions and perhaps destabilize chromatin folding. Here I review recent studies that suggest these potent machines can be 'tamed' by one of several mechanisms: targeting their activity to localized regions, blocking their chromatin binding activity or inhibiting their remodeling activity. PMID- 11943762 TI - Genetic manipulation of insulin signaling, action and secretion in mice. Insights into glucose homeostasis and pathogenesis of type 2 diabetes. AB - Non-insulin-dependent diabetes mellitus (NIDDM) is a complex heterogeneous polygenic disease characterized mainly by insulin resistance and pancreatic beta cell dysfunction. In recent years, several genetically engineered mouse models have been developed for the study of the pathophysiological consequences of defined alterations in a single gene or in a set of candidate diabetogenes. These represent new tools that are providing invaluable insights into NIDDM pathogenesis. In this review, we highlight the lessons emerging from the study of some of the transgenic or knockout mice in which the expression of key actors in insulin signaling, action or secretion has been manipulated. In addition to contributing to our knowledge of the specific roles of individual genes in the control of glucose homeostasis, these studies have made it possible to address several crucial issues in NIDDM that have remained controversial or unanswered for a number of years. PMID- 11943763 TI - Use of stepwise subtraction to comprehensively isolate mouse genes whose transcription is up-regulated during spermiogenesis. AB - We report the isolation of 153 mouse genes whose expression is dramatically up regulated during spermiogenesis. We used a novel variation of the subtractive hybridization technique called stepwise subtraction, wherein the subtraction process is systematically repeated in a stepwise manner. We named the genes thus identified as TISP genes (transcript induced in spermiogenesis). The transcription of 80 of these TISP genes is almost completely specific to the testis. This transcription is abruptly turned on after 17 days of age, when the mice enter puberty and spermiogenesis is initiated. Considering that the most advanced cells present at these stages of spermatogenesis are the spermatids, it is likely that we could isolate most of the spermatid-specific genes. DNA sequencing revealed that about half the TISP genes are novel and uncharacterized genes, confirming the utility of the stepwise subtraction approach for gene discovery. PMID- 11943764 TI - The Drosophila homolog of NTF-2, the nuclear transport factor-2, is essential for immune response. AB - Nuclear transport factor-2 (NTF-2) functions in yeast and mammalian cell culture in targeting proteins into the nucleus. The Drosophila homolog, DNTF-2, is an essential component of the nuclear import machinery, since ntf mutants are lethal. Interestingly, hypomorphic alleles show specific phenotypes. Some are viable, but the number of omatidia in the eye is severely reduced. The immune response in the Drosophila larval fat body is also affected; the three NF kappaB/Rel proteins Dorsal, Dif and Relish do not target to the nucleus after infection, and, consequently, the expression of the anti-microbial peptide genes drosomycin, attacin and drosocin is severely impaired. Hence, in spite of its general requirement in many developmental processes, DNTF-2 has a higher specific requirement in the development of the eye and in the immune response. We also found that DNTF-2 interacts directly with Mbo/DNup88, which does not contain phenylalanine-glycine-rich repeats, but has been shown to function in the import of Rel proteins. PMID- 11943765 TI - Presenilin 1 is required for maturation and cell surface accumulation of nicastrin. AB - Proteolytic processing of amyloid precursor protein generates beta-amyloid (Abeta) peptides that are deposited in senile plaques in brains of aged individuals and patients with Alzheimer's disease. Presenilins (PS1 and PS2) facilitate the final step in Abeta production, the intramembranous gamma secretase cleavage of amyloid precursor protein. Biochemical and pharmacological evidence support a catalytic or accessory role for PS1 in gamma-secretase cleavage, as well as a regulatory role in select membrane protein trafficking. In this report, we demonstrate that PS1 is required for maturation and cell surface accumulation of nicastrin, an integral component of the multimeric gamma secretase complex. Using kinetic labeling studies we show that in PS1(-/-)/PS2(-/ ) cells nicastrin fails to reach the medial Golgi compartment, and as a consequence, is incompletely glycosylated. Stable expression of human PS1 restores these deficiencies in PS1(-/-) fibroblasts. Moreover, membrane fractionation studies show co-localization of PS1 fragments with mature nicastrin. These results indicate a novel chaperone-type role for PS1 and PS2 in facilitating nicastrin maturation and transport in the early biosynthetic compartments. Our findings are consistent with PS1 influencing gamma-secretase processing at multiple steps, including maturation and intracellular trafficking of substrates and component(s) of the gamma-secretase complex. PMID- 11943766 TI - Substitution of a single amino acid switches the tentoxin-resistant thermophilic F1-ATPase into a tentoxin-sensitive enzyme. AB - In contrast to the homologous bacterial and mitochondrial enzymes the chloroplast F(1)-ATPase (CF(1)) is strongly affected by the phytopathogenic inhibitor tentoxin. Based on structural information obtained from crystals of a CF(1) tentoxin co-complex (Groth, G. (2002) Proc. Natl. Acad. Sci. U. S. A. 99, 3464 3468) we have replaced residues betaSer(66) and alphaArg(132) in the alpha(3)beta(3)gamma subcomplex of the thermophilic F(1)-ATPase from Bacillus PS3 by the corresponding residues of the chloroplast ATPase to confer tentoxin sensitivity to the thermophilic enzyme. The mutation alphaArg(132) --> Pro, proposed to relieve steric constraints on tentoxin binding, did not have any significant effect. However, mutation betaSer(66) --> Ala, predicted to provide a crucial hydrogen bond with the inhibitor, resulted in tentoxin inhibition of ATP hydrolysis comparable with the situation found with the chloroplast enzyme. PMID- 11943767 TI - The cytochrome P450 complement (CYPome) of Streptomyces coelicolor A3(2). AB - In the present study we describe the complete cytochrome P450 complement, the "CYPome," of Streptomyces coelicolor A3(2). Eighteen cytochromes P450 (CYP) are described, in contrast to the absence of CYPs in Escherichia coli, and the twenty observed in Mycobacterium tuberculosis. Here we confirm protein identity as cytochromes P450 by heterologous expression in E. coli and measurement of reduced carbon monoxide difference spectra. We also report on their arrangement in the linear chromosome and relatedness to other CYPs in the superfamily. The future development of manipulation of antibiotic pathways and the use of streptomycetes in bioremediation and biotransformations will involve many of the new CYP forms identified here. PMID- 11943768 TI - Mechanisms of FOXC2- and FOXD1-mediated regulation of the RI alpha subunit of cAMP-dependent protein kinase include release of transcriptional repression and activation by protein kinase B alpha and cAMP. AB - We have reported recently that mice overexpressing the forkhead/winged helix transcription factor FOXC2 are lean and show increased responsiveness to insulin due to sensitization of the beta-adrenergic cAMP-PKA(+) pathway and increased levels of the RI alpha subunit of cAMP-dependent protein kinase (PKA) (Cederberg, A., Gronning, L. M., Ahren, B., Tasken, K., Carlsson, P., and Enerback, S. (2001) Cell 106, 563-573). In this present study, we reveal that FOXC2 and a related factor, FOXD1, specifically activate the 1b promoter of the RI alpha gene in adipocytes and testicular Sertoli cells, respectively. By deletional mapping, we discovered two different mechanisms by which the Fox proteins activated expression from the RI alpha 1b promoter. In 3T3-L1 adipocytes, an upstream region represses promoter activity under basal conditions. Bandshift experiments indicate that overexpression of FOXC2 promotes the release of a potential repressor from this region. In Sertoli cells, sequences downstream of the transcription start sites mediate the activating effect of FOXD1, and protein kinase B alpha/Akt1 strongly induces this effect. Furthermore, we show that an inactive FOXD1 mutant lowers the cAMP-mediated induction of the RI alpha 1b reporter construct. In summary, winged helix transcription factors of the FOXC/FOXD families function as regulators of the RI alpha subunit of PKA and may integrate hormonal signals acting through protein kinase B and cAMP in a cell specific manner. PMID- 11943769 TI - Conformation of beta 2-microglobulin amyloid fibrils analyzed by reduction of the disulfide bond. AB - Beta2-microglobulin (beta2-m), a major component of dialysis-related amyloid fibrils, has an intrachain disulfide bond buried inside the native structure. We examined the conformation of beta2-m amyloid fibrils by analyzing the reactivity of the disulfide bond to a reducing reagent, dithiothreitol. Although the disulfide bond in the native structure was highly protected from reduction, the disulfide bonds in the amyloid fibrils prepared at pH 2.5 were progressively reduced at pH 8.5 by 50 mm dithiothreitol. Because beta2-m amyloid fibrils prepared under acidic conditions have been known to depolymerize at a neutral pH, we examined the relation between depolymerization and reduction of the disulfide bond. The results indicate that the disulfide bonds in the amyloid fibrils were protected from reduction, and the reduction occurred during depolymerization. On the other hand, the disulfide bonds of immature filaments, the thin and flexible filaments prepared under conditions of high salt at pH 2.5, were reduced at pH 8.5 more readily than those of amyloid fibrils, suggesting that the disulfide bonds are exposed to the solvent. Taken together, the disulfide bond once exposed to the solvent upon acid denaturation may be progressively buried in the interior of the amyloid fibrils during its formation. PMID- 11943770 TI - Phenotypic spectrum caused by transgenic overexpression of activated Akt in the heart. AB - The serine-threonine kinase, Akt, inhibits cardiomyocyte apoptosis acutely both in vitro and in vivo. However, the effects of chronic Akt activation in the heart are unknown. To address this issue, we generated transgenic mice (TG+) with cardiac-specific expression of a constitutively active mutant of Akt (myr-Akt) driven by the myosin heavy chain-alpha promoter. Three TG+ founders (9-19 weeks) died suddenly with massive cardiac dilatation. Two viable TG+ lines (TG564 and TG20) derived from independent founders demonstrated cardiac-specific transgene expression as well as activation of Akt and p70S6 kinase. TG564 (n = 19) showed cardiac hypertrophy with a heart/body weight ratio 2.3-fold greater than littermates (n = 17, p < 0.005). TG20 (n = 18) had less marked cardiac hypertrophy with a heart/body weight ratio 1.6-fold greater than littermates (n = 17, p < 0.005). Isolated TG564 myocytes were also hypertrophic with surface areas 1.7-fold greater than littermates (p < 0.000001). Echocardiograms in both lines demonstrated concentric hypertrophy and preserved systolic function. After ischemia-reperfusion, TG+ had a 50% reduction in infarct size versus TG- (17 +/- 3% versus 34 +/- 4%, p < 0.001). Thus, chronic Akt activation is sufficient to cause a spectrum of phenotypes from moderate cardiac hypertrophy with preserved systolic function and cardioprotection to massive cardiac dilatation and sudden death. PMID- 11943771 TI - A genomic and proteomic analysis of activation of the human neutrophil by lipopolysaccharide and its mediation by p38 mitogen-activated protein kinase. AB - Bacterial lipopolysaccharide (LPS) evokes several functional responses in the neutrophil that contribute to innate immunity. Although certain responses, such as adhesion and synthesis of tumor necrosis factor-alpha, are inhibited by pretreatment with an inhibitor of p38 mitogen-activated protein kinase, others, such as actin assembly, are unaffected. The aim of the present study was to investigate the changes in neutrophil gene transcription and protein expression following lipopolysaccharide exposure and to establish their dependence on p38 signaling. Microarray analysis indicated expression of 13% of the 7070 Affymetrix gene set in nonstimulated neutrophils, and LPS up-regulation of 100 distinct genes, including cytokines and chemokines, signaling molecules, and regulators of transcription. Proteomic analysis yielded a separate list of up-regulated modulators of inflammation, signaling molecules, and cytoskeletal proteins. Poor concordance between mRNA transcript and protein expression changes was noted. Pretreatment with the p38 inhibitor SB203580 attenuated 23% of LPS-regulated genes and 18% of LPS-regulated proteins by > or = 40%. This study indicates that p38 plays a selective role in regulation of neutrophil transcripts and proteins following lipopolysaccharide exposure, clarifies that several of the effects of lipopolysaccharide are post-transcriptional and post-translational, and identifies several proteins not previously reported to be involved in the innate immune response. PMID- 11943772 TI - Association of Fyn and Lyn with the proline-rich domain of glycoprotein VI regulates intracellular signaling. AB - The glycoprotein VI (GPVI)-Fc receptor (FcR) gamma-chain complex, a key activatory receptor for collagen on platelet surface membranes, is constitutively associated with the Src family kinases Fyn and Lyn. Molecular cloning of GPVI has revealed the presence of a proline-rich domain in the sequence of GPVI cytoplasmic tail which has the consensus for interaction with the Src homology 3 (SH3) domains of Fyn and Lyn. A series of in vitro experiments demonstrated the ability of the SH3 domains of both Src kinases to bind the proline-rich domain of GPVI. Furthermore, depletion of the proline-rich domain in GPVI (Pro(-)-GPVI) prevented binding of Fyn and Lyn and markedly reduced phosphorylation of FcR gamma-chain in transiently transfected COS-7 cells, but did not affect the association of the gamma-chain with GPVI. Jurkat cells stably transfected with wild type GPVI show robust increases in tyrosine phosphorylation and intracellular Ca2+ in response to the snake venom convulxin that targets GPVI. Importantly, convulxin is not able to activate cells transfected with Pro(-) GPVI, even though the association with the immunoreceptor tyrosine-based activation motif-containing chains is maintained. These findings demonstrate that the proline-rich domain of GPVI mediates the association with Fyn/Lyn via their SH3 domain and that this interaction initiates activation signals through GPVI. PMID- 11943773 TI - Identification of the regulatory elements of the human von Willebrand factor for binding to platelet GPIb. Importance of structural integrity of the regions flanked by the CYS1272-CYS1458 disulfide bond. AB - In vitro platelet glycoprotein Ib (GPIb) binding of the human von Willebrand factor (VWF) increases markedly by exogenous modulators such as ristocetin or botrocetin, and the binding does not occur in normal circulation. GPIb binding sites have been assigned in the VWF A1 domain, which consists of a disulfide loop Cys1272(509)-Cys1458(695) where amino acid residues are numbered from the starting methionine as +1. The previous numbering from the N-terminal Ser of the mature processed VWF is indicated in parentheses. In contrast, several gain-of function mutations have been found in two regions comprised of the disulfide loop and its N- and C-terminal flanking regions. In this study, Cys1222(459) Tyr1271(508), Gln1238(475)-Tyr1271(508), Glu1260(497)-Tyr1271(508), and Asp1459(696)-Asp1472(709) were sequentially deleted of full-length multimeric recombinant VWF. Deletions at either side resulted in normal GPIb binding, indicating that the flanking regions are not GPIb binding sites. However, the addition of a mutation at Arg1308(545) on each deletion mutant resulted in spontaneous GPIb binding without requiring modulators, suggesting that both regions are important for the inhibition of GPIb binding. Spontaneous binding was completely inhibited by monoclonal antibodies that recognize the GPIb binding sites. Interestingly, mutant proteins with N-terminal but not C-terminal deletions lost binding to monoclonal antibodies B328, B710, and 23C7, which selectively inhibit ristocetin-induced GPI binding. Their epitopes were found at His1268(505) or Asp1269(506). The crystallographic structure of the A1 domain suggests that GPIb binding is influenced by the molecular interface between the two regions and that the antibody binding to the interface inhibits binding. PMID- 11943774 TI - The rod cGMP-phosphodiesterase beta-subunit promoter is a specific target for Sp4 and is not activated by other Sp proteins or CRX. AB - The beta-subunit of cGMP-phosphodiesterase (beta-PDE) is a key protein in phototransduction expressed exclusively in rod photoreceptors. It is necessary for visual function and for structural integrity of the retina. beta-PDE promoter deletions showed that the -45/-23 region containing a consensus Crx-response element (CRE) was necessary for low level transcriptional activity. Overexpressed Crx modestly transactivated this promoter in 293 human embryonic kidney cells; however, mutation of CRE had no significant effect on transcription either in transfected Y79 retinoblastoma cells or Xenopus embryonic heads. Thus, Crx is unlikely to be a critical beta-PDE transcriptional regulator in vivo. Interestingly, although the beta/GC element (-59/-49) binds multiple Sp transcription factors in vitro, only Sp4, but not Sp1 or Sp3, significantly enhanced beta-PDE promoter activity. Thus, the Sp4-mediated differential activation of the beta-PDE transcription defines the first specific Sp4 target gene reported to date and implies the importance of Sp4 for retinal function. Further extensive mutagenesis of the beta-PDE upstream sequences showed no additional regulatory elements. Although this promoter lacks a canonical TATA box or Inr element, it has the (T/A)-rich beta/TA sequence located within the -45/-23 region. We found that it binds purified TBP and TFIIB in gel mobility shift assays with cooperative enhancement of binding affinity. PMID- 11943775 TI - Active site residues of cyclophilin A are crucial for its signaling activity via CD147. AB - Cyclophilin A (CyPA), a ubiquitously distributed intracellular protein, is a peptidylprolyl cis-trans-isomerase and the major target of the potent immunosuppressive drug cyclosporin A. Although expressed predominantly as an intracellular molecule, CyPA is secreted by cells in response to inflammatory stimuli and is a potent neutrophil and eosinophil chemoattractant in vitro and in vivo. The mechanisms underlying CyPA-mediated signaling and chemotaxis are unknown. Here, we identified CD147 as a cell surface receptor for CyPA and demonstrated that CD147 is an essential component in the CyPA-initiated signaling cascade that culminates in ERK activation. Both signaling and chemotactic activities of CyPA depended also on the presence of heparans, which served as primary binding sites for CyPA on target cells. The proline 180 and glycine 181 residues in the extracellular domain of CD147 were critical for signaling and chemotactic activities mediated by CD147. Also crucial were active site residues of CyPA, because rotamase-defective CyPA mutants failed to initiate signaling events. These results establish cyclophilins as natural ligands for CD147 and suggest an unusual, rotamase-dependent mechanism of signaling. PMID- 11943776 TI - The gibberellin 20-oxidase of Gibberella fujikuroi is a multifunctional monooxygenase. AB - The genes for gibberellin (GA) biosynthesis are clustered in the fungus Gibberella fujikuroi. In addition to genes encoding a GA-specific geranylgeranyl diphosphate synthase and a bifunctional ent-copalyl diphosphate/ent-kaurene synthase, the cluster contains four cytochrome P450 monooxygenase genes (P450-1, 2, -3, -4). Recently it was shown that P450-4 and P450-1 encode multifunctional enzymes catalyzing the three oxidation steps from ent-kaurene to ent-kaurenoic acid and the four oxidation steps from ent-kaurenoic acid to GA14, respectively. Here we describe the functional analysis of the P450-2 gene by gene disruption and by expressing the gene in a mutant that lacks the entire GA biosynthesis gene cluster. Mutants in which P450-2 is inactivated by the insertion of a large piece of DNA accumulated GA14 and lacked biosynthetically more advanced metabolites, indicating that the gene encodes a 20-oxidase. This was confirmed by incubating lines containing P450-2 in the absence of the other GA biosynthesis genes with isotopically labeled substrates. The P450-2 gene product oxidized the 3beta hydroxylated intermediate, GA14, and its non-hydroxylated analogue GA12 to GA4 and GA9, respectively. Expression of P450-2 is repressed by high amounts of nitrogen in the culture medium but is not affected by the presence of biosynthetically advanced GAs, i.e. there is no evidence for feedback regulation. The fact that the GA 20-oxidase is a cytochrome P450 monooxygenase in G. fujikuroi and not a 2-oxoglutarate-dependent dioxygenase as in plants, together with the significant differences in regulation of gene expression, are further evidence for independent evolution of the GA biosynthetic pathways in plants and fungi. PMID- 11943777 TI - The paired-like homeodomain protein, Arix, mediates protein kinase A-stimulated dopamine beta-hydroxylase gene transcription through its phosphorylation status. AB - The homeodomain transcription factor Arix/Phox2a plays a critical role in the specification of noradrenergic neurons by inducing the expression of dopamine beta-hydroxylase (DBH), the terminal enzyme for noradrenaline biosynthesis. In reporter assays, Arix together with activation of cAMP-dependent protein kinase (PKA) potentiates DBH gene transcription. We have evaluated whether post translational modification of Arix regulates PKA-mediated DBH gene transcription. We found that Arix is constitutively phosphorylated in vivo at the basal level and that the phosphorylation level is substantially decreased upon stimulation of the PKA pathway. The change in the Arix phosphorylation state coincides with DNA binding activity of Arix. Treatment of cells with forskolin results in a robust enhancement of the DNA binding of Arix, which is reversed by treatment with serine/threonine and tyrosine phosphatase inhibitors. Consistent with the DNA binding activity of Arix, treatment of cultured cells with phosphatase inhibitors diminishes transcriptional activation with Arix plus forskolin. Amino acid analysis demonstrates the presence of phosphoserine within Arix. The results collectively suggest that dephosphorylation of Arix is a necessary event to fully activate PKA-mediated DBH transcription. Thus, the present study demonstrates that Arix can integrate extrinsic signals through post-translational modification, regulating DBH gene transcription in response to activation of the PKA pathway. PMID- 11943778 TI - Molecular cloning and characterization of chondroitin polymerase from Escherichia coli strain K4. AB - Escherichia coli strain K4 produces the K4 antigen, a capsule polysaccharide consisting of a chondroitin backbone (GlcUA beta(1-3)-GalNAc beta(1-4))(n) to which beta-fructose is linked at position C-3 of the GlcUA residue. We molecularly cloned region 2 of the K4 capsular gene cluster essential for biosynthesis of the polysaccharide, and we further identified a gene encoding a bifunctional glycosyltransferase that polymerizes the chondroitin backbone. The enzyme, containing two conserved glycosyltransferase sites, showed 59 and 61% identity at the amino acid level to class 2 hyaluronan synthase and chondroitin synthase from Pasteurella multocida, respectively. The soluble enzyme expressed in a bacterial expression system transferred GalNAc and GlcUA residues alternately, and polymerized the chondroitin chain up to a molecular mass of 20 kDa when chondroitin sulfate hexasaccharide was used as an acceptor. The enzyme exhibited apparent K(m) values for UDP-GlcUA and UDP-GalNAc of 3.44 and 31.6 microm, respectively, and absolutely required acceptors of chondroitin sulfate polymers and oligosaccharides at least longer than a tetrasaccharide. In addition, chondroitin polymers and oligosaccharides and hyaluronan polymers and oligosaccharides served as acceptors for chondroitin polymerization, but dermatan sulfate and heparin did not. These results may lead to elucidation of the mechanism for chondroitin chain synthesis in both microorganisms and mammals. PMID- 11943779 TI - CREB-binding protein/p300 co-activation of crystallin gene expression. AB - Although some of the transcription factors that are required for expression of crystallins during lens development have been identified, the molecular interactions that contribute to enhanced crystallin expression are not yet well defined. In this study, we designed experiments to test whether the co-activators CREB-binding protein (CBP) and/or p300 interact with c-Maf, Prox-1, or Sox-1 to enhance transcription of crystallin genes. Promoter regions from the mouse alphaA , betaB2-, and gammaF-crystallin genes were linked to a luciferase reporter. Expression of c-Maf transactivated each of these promoters. Of particular interest, co-expression of CBP or p300 with c-Maf was found to synergistically co activate each promoter. CBP and p300 were less effective or ineffective at co activation with Prox-1 or Sox-1. Co-immunoprecipitation and mammalian two-hybrid experiments revealed that CBP and p300 bind to c-Maf and Prox-1 but not to Sox-1. The co-activation of c-Maf by CBP/p300 requires histone acetyltransferase activity. Our results suggest that c-Maf recruits CBP and/or p300 to crystallin promoters leading to up-regulation of crystallin gene expression through localized histone acetylation and consequent chromatin re-modeling. In a promoter specific fashion, co-activation can be modulated by Prox-1 and/or Sox-1. This modulation may help to specify the endogenous levels of crystallin gene expression. PMID- 11943780 TI - Peg3/Pw1 is a mediator between p53 and Bax in DNA damage-induced neuronal death. AB - Neuronal cell death after DNA damage requires p53 and Bax, but the mechanism by which p53 activation leads to Bax translocation and cell death in neurons is not known. We report here that Peg3/Pw1 is up-regulated after DNA damage in cortical neurons in a p53-dependent manner. Overexpression of Peg3/Pw1 leads to decreased neuronal viability. The deleterious effect of Peg3/Pw1 on neuronal survival is abrogated by deletion of either p53 or Bax, indicating an essential role for both in Peg3/Pw1-mediated neuronal death. Moreover, overexpression of a Peg3/Pw1 dominant negative protein inhibits Bax translocation and neuronal cell death after DNA damage. These findings implicate Peg3/Pw1 as a mediator between p53 and Bax in a neuronal cell death pathway activated by DNA damage. PMID- 11943781 TI - Stabilization of the biotinoyl domain of Escherichia coli acetyl-CoA carboxylase by interactions between the attached biotin and the protruding "thumb" structure. AB - We previously reported (Chapman-Smith, A., Forbes, B. E., Wallace, J. C., and Cronan, J. E., Jr. (1997) J. Biol. Chem. 272, 26017-26022) that the biotinylated (holo) species of the biotin carboxyl carrier protein (BCCP) biotinoyl domain is much more resistant to chemical modification and proteolysis than the unbiotinylated (apo) form. We hypothesized that the increased stability was due to a conformational change engendered by interaction of the domain with biotin protein ligase, the enzyme that attaches the biotin moiety. We now report that a BCCP-87 species to which the biotin moiety was attached by chemical acylation rather than by biotin protein ligase showed the characteristically greater stability of the holo biotinoyl domain. This result demonstrates that our hypothesis was incorrect; the attached biotin is solely responsible for the increased stability. The bacterial and chloroplast multisubunit acetyl-CoA carboxylases are unusual in that the highly symmetrical and conserved structure of the biotinoyl domain of the BCCP subunit is disrupted by a structured loop called the "thumb" that protrudes from body of the domain. Prior structural work showed that the thumb interacts with uriedo ring of the attached biotin moiety. We have tested whether the thumb-biotin interactions are responsible for the greater holo form stability by examination of two BCCP-87 species that lack the thumb. These BCCP species were produced in both the apo and holo forms, and their sensitivities to trypsin digestion were compared. The holo forms of these proteins were found to be only marginally more stable than their apo forms and much more sensitive to trypsin digestion than the wild type holo-BCCP-87. Therefore, removal of the thumb has an effect similar to lack of biotinylation, indicating that thumb-biotin interactions are responsible for most (but not all) of the increased stability of the holo biotinoyl domain. In the course of these experiments we demonstrated that treatment of Escherichia coli with the peptide deformylase inhibitor, actinonin, results in the expected (but previously unreported) accumulation of an N-formylated protein species. PMID- 11943782 TI - Global and specific translational control by rapamycin in T cells uncovered by microarrays and proteomics. AB - Rapamycin has been shown to affect translation. We have utilized two complementary approaches to identify genes that are predominantly affected by rapamycin in Jurkat T cells. One was to compare levels of polysome-bound and total RNA using oligonucleotide microarrays complementary to 6,300 human genes. Another was to determine protein synthesis levels using two-dimensional PAGE. Analysis of expression changes at the polysome-bound RNA levels showed that translation of most of the expressed genes was partially reduced following rapamycin treatment. However, translation of 136 genes (6% of the expressed genes) was totally inhibited. This group included genes encoding RNA-binding proteins and several proteasome subunit members. Translation of a set of 159 genes (7%) was largely unaffected by rapamycin treatment. These genes included transcription factors, kinases, phosphatases, and members of the RAS superfamily. Analysis of [(35)S]methionine-labeled proteins from the same cell populations using two-dimensional PAGE showed that the integrated intensity of 111 of 830 protein spots changed in rapamycin-treated cells by at least 3-fold (70 increased, 41 decreased). We identified 22 affected protein spots representing protein products of 16 genes. The combined microarray and proteomic approach has uncovered novel genes affected by rapamycin that may be involved in its immunosuppressive effect and other genes that are not affected at the level of translation in a context of general inhibition of cap-dependent translation. PMID- 11943783 TI - Evaluation of critical structural elements of UDP-sugar substrates and certain cysteine residues of a vertebrate hyaluronan synthase. AB - The hyaluronan (HA) synthases catalyze the addition of two different monosaccharides from UDP-sugar substrates to the linear heteropolysaccharide chain. To accomplish this task, the HA synthases must be able to bind and to transfer from both UDP-sugar substrates. Until now, it has been impossible to distinguish between these two abilities. We have created a mutant of xlHAS1, a HA synthase from Xenopus laevis, that allows for the examination of the enzyme's ability to bind substrate only. The ability of different compounds to protect the xlHAS1(C337S) mutant enzyme from loss of activity due to treatment with N ethylmaleimide, a cysteine-modifying reagent, yields information on the relative affinity of a variety of nucleotides and nucleotide-sugars. We have observed that the substrate binding selectivity is more relaxed than the specificity of catalytic transfer. The only attribute that appears to be absolutely required for binding is a nucleotide containing two phosphates complexed with magnesium ion. The role of certain cysteine residues in catalysis was also evaluated. Cys307 of xlHAS1 may play a role in catalysis or in maintaining structure. Mutation of Cys337 raises the UDP-GlcUA Michaelis constant (K(m)), suggesting that this residue participates in UDP-GlcUA substrate binding or in catalytic complex formation. PMID- 11943784 TI - Hypoxia-inducible factor 1 activation by aerobic glycolysis implicates the Warburg effect in carcinogenesis. AB - Cancer cells display high rates of aerobic glycolysis, a phenomenon known historically as the Warburg effect. Lactate and pyruvate, the end products of glycolysis, are highly produced by cancer cells even in the presence of oxygen. Hypoxia-induced gene expression in cancer cells has been linked to malignant transformation. Here we provide evidence that lactate and pyruvate regulate hypoxia-inducible gene expression independently of hypoxia by stimulating the accumulation of hypoxia-inducible Factor 1alpha (HIF-1alpha). In human gliomas and other cancer cell lines, the accumulation of HIF-1alpha protein under aerobic conditions requires the metabolism of glucose to pyruvate that prevents the aerobic degradation of HIF-1alpha protein, activates HIF-1 DNA binding activity, and enhances the expression of several HIF-1-activated genes including erythropoietin, vascular endothelial growth factor, glucose transporter 3, and aldolase A. Our findings support a novel role for pyruvate in metabolic signaling and suggest a mechanism by which high rates of aerobic glycolysis can promote the malignant transformation and survival of cancer cells. PMID- 11943785 TI - Comparison of human TRPC3 channels in receptor-activated and store-operated modes. Differential sensitivity to channel blockers suggests fundamental differences in channel composition. AB - Capacitative calcium entry or store-operated calcium entry in nonexcitable cells is a process whereby the activation of calcium influx across the plasma membrane is signaled by depletion of intracellular calcium stores. Transient receptor potential (TRP) proteins have been proposed as candidates for store-operated calcium channels. Human TRPC3 (hTRPC3), an extensively studied member of the TRP family, is activated through a phospholipase C-dependent mechanism, not by store depletion, when expressed in HEK293 cells. However, store depletion by thapsigargin is sufficient to activate hTRPC3 channels when expressed in DT40 avian B-lymphocytes. To gain further insights into the differences between hTRPC3 channels generated in these two expression systems and further understand the role of hTRPC3 in capacitative calcium entry, we examined the effect of two well characterized inhibitors of capacitative calcium entry, Gd3+ and 2 aminoethoxydiphenyl borane (2APB). We confirmed that in both DT40 cells and HEK293 cells, 1 microm Gd3+ or 30 microm 2APB completely blocked calcium entry due to receptor activation or store depletion. In HEK293 cells, 1 microm Gd3+ did not block receptor-activated hTRPC3-mediated cation entry, whereas 2APB had a partial (approximately 60%) inhibitory effect. Interestingly, store-operated hTRPC3-mediated cation entry in DT40 cells was also partially inhibited by 2APB, whereas 1 microm Gd3+ completely blocked store-operated hTRPC3 activity in these cells. Furthermore, the sensitivity of store-operated hTRPC3 channels to Gd3+ in DT40 cells was similar to the endogenous store-operated channels, with essentially 100% block of activity at concentrations as low as 0.1 microm. Finally, Gd3+ has a rapid inhibitory effect when added to fully developed hTRPC3 mediated calcium entry, suggesting a direct action of Gd3+ on hTRPC3 channels. The distinct action of these inhibitors on hTRPC3-mediated cation entry in these two cell types may result from their different modes of activation and may also reflect differences in basic channel structure. PMID- 11943786 TI - New regulators of drug sensitivity in the family of yeast zinc cluster proteins. AB - The Gal4p family of yeast zinc cluster proteins comprises over 50 members that are putative transcriptional regulators. For example, Pdr1p and Pdr3p activate multidrug resistance genes by binding to pleiotropic drug response elements (PDREs) found in promoters of target genes such as PDR5, encoding a drug efflux pump involved in resistance to cycloheximide. However, the role of many zinc cluster proteins is unknown. We tested a panel of strains carrying deletions of zinc cluster genes in the presence of various drugs. One deletion strain (Deltardr1) was resistant to cycloheximide, whereas eight strains showed sensitivity to the antifungal ketoconazole or cycloheximide. Unnamed zinc cluster genes identified in our screen were called RDS for regulators of drug sensitivity. RNA levels of multidrug resistance genes such as PDR16, SNQ2, and PDR5 were decreased in many deletion strains. For example, cycloheximide sensitivity of a Deltastb5 strain was correlated with decreased RNA levels and promoter activity of the PDR5 gene. We tested if activation of PDR5 is mediated via a PDRE by inserting this DNA element in front of a minimal promoter linked to the lacZ gene. Strikingly, activity of the reporter was decreased in a Deltastb5 strain. The purified DNA binding domain of Stb5p bound to a PDRE in vitro. Mutations in the PDRE known to affect binding of Pdr1p/Pdr3p showed similar effects when assayed with Stb5p. These results strongly suggest that Stb5p is a transcriptional activator of multidrug resistance genes. Thus, we have identified new regulators of drug sensitivity in the family of zinc cluster proteins. PMID- 11943787 TI - The minor subunit splice variants, H2b and H2c, of the human asialoglycoprotein receptor are present with the major subunit H1 in different hetero-oligomeric receptor complexes. AB - The hepatic asialoglycoprotein receptor (ASGP-R) is an endocytic receptor that mediates the internalization of desialylated glycoproteins and their delivery to lysosomes. The human ASGP-R is a hetero-oligomeric complex composed of H1 and H2 subunits. There are three naturally occurring H2 splice variants, designated H2a, H2b, and H2c, although the expression of the H2c protein had not been reported. Following deglycosylation of purified ASGP-R, we detected the H2b and H2c proteins in HepG2 and HuH-7 hepatoma cells, using an antibody directed against a COOH-terminal peptide common to all H2 isoforms (anti-H2-COOH) and another antibody against a 19-amino acid cytoplasmic insert found only in H2b (anti-H2 Cyto19). H1 and both H2b and H2c were co-purified by affinity chromatography, using asialo-orosomucoid (ASOR)-, anti-H1-, or anti-H2-COOH-Sepharose, whereas only H1 and H2b were immunoprecipitated with anti-H2-Cyto19. These results indicate that H2b and H2c are not present in the same ASGP-R complexes with H1. Similar to the H2b isoform, H2c was also palmitoylated, indicating that the 19 residue cytoplasmic insert does not regulate palmitoylation. Stably transfected SK-Hep-1 cell lines expressing ASGP-R complexes containing H1 and either H2b or H2c had similar binding affinities for ASOR and endocytosed and degraded ASOR at similar rates. The pH dissociation profiles of ASOR.ASGP-R complexes were also identical for complexes containing either H2b or H2c. We conclude that the H2b and H2c isoforms are both functional but are not present with H1 in the same hetero-oligomeric ASGP-R complexes. This structural difference between two functional subpopulations of ASGP-Rs may provide a molecular basis for the existence of two different pathways, designated State 1 and State 2, by which several types of recycling receptors mediate endocytosis. PMID- 11943789 TI - Revised estimates of the maternal age specific live birth prevalence of Down's syndrome. AB - OBJECTIVES: To revise the estimates of maternal age specific live birth prevalence of Down's syndrome in the absence of antenatal screening and selective termination using newly available data. SETTING AND DESIGN: Data were used from the National Down Syndrome Cytogenetic Register (NDSCR), which contains information on nearly all antenatally or postnatally diagnosed cases of Down's syndrome in which a karyotype was confirmed between 1989 and 1998 in England and Wales. It is the largest single series of data on the prevalence of Down's syndrome. RESULTS AND CONCLUSION: The prevalence does not continue increasing at an increasing rate with age above age 45 as has been previously assumed. Above this age the rate of increase declines with increasing age. The overall age pattern is sigmoidal. A new logit logistic model is proposed which fits the data well. The risk of a Down's syndrome live birth is given by: risk=1/(1+exp(7.330 4.211/(1+exp(-0.282x(age-37.23))))). PMID- 11943790 TI - Psychiatric morbidity and screening for colorectal cancer. AB - BACKGROUND: Several studies have shown that faecal occult blood (FOB) screening reduces mortality from colorectal cancer. However, concern has been expressed that health screening may have adverse psychological effects, particularly for the group returning false positive tests. OBJECTIVES: To evaluate any adverse psychological effects associated with faecal occult blood screening. SETTING: Randomised controlled trial of faecal occult blood screening for colorectal cancer. METHODS: Psychiatric morbidity was measured, using the general health questionnaire (GHQ) before and 3 months after the offer of screening for colorectal cancer with FOB testing. Scores were related to acceptance of the screening test. A smaller cohort, who had returned positive FOB tests, had anxiety levels measured, using the Spielberger anxiety inventory (SAI), at different times during screening, investigation, and follow up. RESULTS: A GHQ was sent to 2184 subjects before the offer of screening, and 1541 (70.6%) were returned. Of the 1693 subjects offered the GHQ 3 months after the offer of screening, 1303 (77%) returned it. A GHQ score of 5 or more, indicating possible psychiatric morbidity, was present in 454 subjects (29.5%) before screening and in 386 (29.6%) subjects 3 months after screening (NS). Of the 454 subjects who scored 5 or more, 241 (53.1%) accepted screening and 213 (46.9%) refused. A total of 1081 subjects scored less than 5, and of these 521 (48.2%) accepted screening and 560 (51.8%) refused (NS). Anxiety scores were measured in 100 test positive patients and were highest after notification of a positive test and before investigation by colonoscopy. In patients with false positive results, scores fell the day after colonoscopy and remained low 1 month later. CONCLUSIONS: The receipt of a screening test does not cause sustained anxiety and the existence of psychiatric morbidity is not a factor affecting a person's decision to accept or refuse a screening test for colorectal cancer. PMID- 11943791 TI - Does individual programme size affect screening performance? Results from the United Kingdom NHS breast screening programme. AB - OBJECTIVE: The size (number of women screened) of the 95 individual NHS breast screening programmes (NHSBSPs) varies by a factor of 10. This study investigates the impact of size on the performance of individual programmes. METHOD: Data were collated from the 95 United Kingdom screening programmes on the standard statistical returns for the past 5 years (1 April 1995-31 March 2000). Additional information was obtained from questionnaires. The number of women screened between 1 April 1999 and 31 March 2000 determined the size of a programme. The bottom 25% were defined as small, the middle 50% as medium, and the top 25% as large. On average large programmes screened about four times as many women as small programmes and medium programmes about twice as many. Performance was evaluated using cancer detection rates, referral rates for assessment, and positive predictive value (PPV) of assessment using PPV referral diagrams. RESULTS: The performance of smaller programmes was shown to be marginally poorer than medium and large sized programmes in that they detected fewer cancers and had a lower PPV. The smallest 25% of programmes had an invasive cancer detection rate 13% less than the medium and large programmes. However, if these programmes had an equivalent detection rate to the medium/large programmes the national detection rate would only increase by about 2%. This is because the 75% of programmes described as medium and large screen about 90% of all women. It is therefore important to place the clinical importance of these findings in context when considering any envisaged possible solutions. CONCLUSIONS: Although the performance of smaller programmes was shown to be poorer than that of the larger programmes, it is not clear from this study exactly why this is so. A likely contributory factor based on experience of evaluating the NHSBSP is that performance problems in larger programmes have been easier to detect by quality assurance staff. The size of the small programmes and the few screen detected cancers (and inherent statistical instability in detection rates) mean that problems are difficult to identify. As a consequence small programmes which are genuinely performing marginally below specific standards are likely to receive less attention than larger programmes, and even under close scrutiny the causes are less likely to be found. PMID- 11943792 TI - An investigation into the effect of extending routine mammographic screening to older women in the United Kingdom on the time it takes to screen. AB - OBJECTIVE: To compare the time it takes to screen women aged 65-69 with women age 50-64. SETTING: Screening centres participating in the Department of Health funded demonstration study of extending breast screening to women aged 65-69. METHODS: Consecutive women of all ages were timed during November 1999 to February 2000, until about 50 women aged 65-69 had been screened at each of the sites. For each woman screened, her age, whether she had been screened before, the times when she checked in, started to get undressed, started screening, finished screening, and when she left the screening unit were recorded. The radiographers also recorded any difficulties associated with screening any of the women in the survey. RESULTS: The total mean time spent at the centres and the time being screened was not significantly different between the two age groups overall or within each of the demonstration sites (p>0.05). The distribution of screen time was similar between the age groups within each of the sites. The proportion of screens reported as difficult by the radiographers differed between sites, with two out of the three sites reporting more problems screening older women. CONCLUSIONS: Despite some radiographers reporting more difficulties associated with screening older women, screening a woman aged 65-69 was found, on average, to take no longer than screening a women aged 50-64 years. Thus, when the NHS breast screening programme is extended to include older women, the same time interval for an appointment can be scheduled regardless of the woman's age. PMID- 11943793 TI - Interval breast cancers in New South Wales, Australia, and comparisons with trials and other mammographic screening programmes. AB - OBJECTIVE: To present information on 1st year interval breast cancer from the New South Wales mammographic screening programme and to compare with published results from trials and services. SETTING: New South Wales data were derived from a population based biennial mammographic screening programme, which achieved statewide coverage in 1995. Women aged 50-69 years screened during 1995-7 were included. METHODS: Bilateral two view mammography with reading by two radiologists is used for biennial screening examinations. Interval cancers were detected by the screening programme and by linkage with the statewide cancer registry. In situ carcinoma was excluded. Incidence of interval cancer was estimated as a proportion of the expected underlying incidence of breast cancer. Comparative data were derived from the published literature and meta-analyses were performed. RESULTS: Although randomised trials of screening have a proportional incidence by meta-analysis of 19% (95% confidence interval (95% CI) 12% to 25%), service studies yield a proportional incidence by meta-analysis of 27% (95% CI 25% to 30%), and more than half report proportional incidences greater than 25%. In the New South Wales mammographic screening programme the proportional incidence of interval breast cancer was 33% (95% CI 29% to 38%) for ages 50-59 years, 28% (95% CI 24% to 32%) for ages 60-69 years, and 31% (95% CI 28% to 34%) for ages 50-69 years combined. Proportional incidence in the New South Wales programme for ages 50-69 years was not significantly different from the rate for service studies by meta-analysis. CONCLUSIONS: Effectiveness of mammographic screening for reducing mortality from breast cancer needs to be examined relative to rates of interval cancer from actual service situations as trials may concentrate resources and expertise in ways which may be less replicable in routine delivery of the service. PMID- 11943794 TI - Early cervical cancer rescreening. AB - INTRODUCTION: The extent of early cervical rescreening, defined as rescreening earlier than the recommended interval for a given initial test result, is difficult to determine mainly because the testing history of women is incomplete, especially with newer population based screening registers. METHODS: Estimation of early rescreening is based on analysis of a 1 month cohort of women (February 1997) recorded on the New South Wales Pap Test Register (PTR) who initially tested negative and who had no recorded history of a positive test result. For the purposes of estimating early rescreening rates and sources of multiple screening occurring within the recommended 2 yearly screening interval for New South Wales, the cohort excluded the estimated proportion of women with an unrecorded history of a positive result. Approaches to exclusion were different for women with a history of a high grade result (CIN2 or higher) or a low grade result (CIN1 or lower). RESULTS: Characteristic rescreening peaks occurred at 6, 12, 18, 24 and 27 months according to negative result category. The rescreening peak at 27 months illustrates the effect of the PTR late reminder system. After adjusting the cohort for estimated proportions of women with a history of a lesion, the number of women estimated to have rescreened early was approximately 156 000 over 1997-8, and the early rescreening rate was estimated as 15.3% of women who have a Pap test. A feasible target for reducing rescreening through service provider interventions was estimated to be a reduction of 7.4%, based on reducing the overscreening proportion from 15.3% to 7.9% through truncating characteristic rescreening peaks to background levels of rescreening. This represents just under 200 000 screens that could be performed over 2 years on New South Wales women who are underscreened or unscreened without incurring additional costs due to screening. CONCLUSIONS: Reasonable estimates of early cervical rescreening can be derived but some assumptions in estimating the proportion of women with a positive test result history may be needed particularly if a screening programme is new and universal recording of screening data is only recent. Characteristic peaks, as departures from a background of random noise in time plots of rescreening indicate that a component of early rescreening is systemic and the chief source of such variation in screening behaviour would be service providers. PMID- 11943795 TI - A randomised trial of population screening for melanoma. AB - OBJECTIVES: Melanoma is a significant cause of morbidity and mortality worldwide and incidence is increasing. Survival after treatment is inversely related to the thickness of the tumour at diagnosis. Population screening has the potential to reduce mortality but there is no conclusive evidence of benefit. Such evidence can come best from a randomised trial. Here we describe the design of a community based randomised trial of a population screening programme for melanoma in Queensland, Australia and early results of the first phase of the trial. METHODS: A total of 44 communities (aggregate population 560 000 adults aged 30 years or more) will be randomised to receive either a community based screening programme for 3 years or normal practice. The screening programme promotes thorough skin self examination and whole body skin examination by a doctor and provides open access skin cancer screening clinics. In its first phase, the trial is underway in nine intervention and nine control communities. The primary outcome measure is mortality from melanoma during 15 years of follow up. RESULTS: The first phase of the trial has shown the feasibility of implementing a population skin screening programme including regular skin cancer screening clinics, and has shown the strong support of communities and doctors for the programme. There has been a significant 2.5-fold increase in participation in screening in the intervention communities in this first phase after the first 12 months of the trial and no significant increase in participation in screening in control communities during this period. CONCLUSIONS: The design of a community based randomised trial of screening for melanoma has been successfully peer reviewed and the intervention has been shown to be feasible in practice. This randomised trial may be one of the last opportunities to develop the evidence required for public health recommendations for population screening for melanoma. PMID- 11943796 TI - An interesting byproduct of screening: assessing the effect of HRT on arterial calcification in the female breast. AB - Arterial calcification in the female breast (BAC) is a common but unreported finding on breast screening mammograms. Accumulating evidence suggests that BAC may have importance as a marker for generalised vascular disease. In this study, we have assessed the influence of HRT usage on the prevalence of BAC in 4400 women undergoing mammography for breast cancer screening. Significantly increased levels of breast arterial calcification were observed in women who had never used HRT compared with a group of women presently using HRT. PMID- 11943797 TI - Probabilities of progression of aortic aneurysms: estimates and implications for screening policy. AB - BACKGROUND: Screening for abdominal aortic aneurysm, and intervention with elective repair, can reduce the incidence of aneurysmal rupture by a half. If a screening programme is implemented, it is essential to determine appropriate follow up intervals for rescreening. This paper estimates probabilities of progression growth of aortic diameter to provide evidence for this. METHODS: Data were taken from 2342 men aged 65-80 screened in the Chichester randomised control trial, who have been followed up for an average of 11 years. Aortic diameter was modelled as a Markov process with four categories: <30 mm (normal), 30-44 mm, 45 54 mm, and > or =55 mm. Estimates of the probabilities of progressing to each higher category were obtained. RESULTS: The probabilities of progression increased with greater initial aortic diameter. The estimated rates/year were 0.018 (95% confidence interval 0.014 to 0.023), 0.16 (0.12 to 0.20), and 0.49 (0.35 to 0.70) respectively for moving up one category. The probabilities of moving from <30 mm to > or =55 mm were estimated as 1% in 5 years and 12% in 15 years, while the corresponding figures for moving from 45-54 mm to > or =55 mm were 91% and 99%. There were differences in rates of progression according to age, with men over 70 years having rates about three times those of men under 70. CONCLUSIONS: It seems unnecessary to follow up men with normal aortic diameter as they experience a low probability of reaching criteria for surgery even within 15 years. However, follow up intervals should be progressively shorter for those with greater aortic diameter, especially in those aged over 70. Active follow up, for example every 3 months, is appropriate for men with an aortic diameter of 45 54 mm. PMID- 11943798 TI - Assigning risk for Smith-Lemli-Opitz syndrome as part of 2nd trimester screening for Down's syndrome. AB - OBJECTIVES: To design a reliable model in the context of prenatal screening for assigning the risk in an individual pregnancy of Smith-Lemli-Opitz syndrome (SLOS) and assess its performance. SETTING: A 2nd trimester screening programme for Down's syndrome that measures unconjugated estriol (uE3) along with other serum markers. METHODS: Development of individual risk estimates with a trivariate model incorporating measurements of maternal serum uE3, alpha fetoprotein (AFP), and human chorionic gonadotropin (hCG) in both SLOS and unaffected pregnancies. RESULTS: Population parameters were computed for the three analytes, as were pairwise correlation coefficients and truncation limits, based on an unbiased collection of 29 affected pregnancies. Published parameters were used for unaffected pregnancies. With a cut off level of risk of 1:50, 62% of SLOS pregnancies can be detected by initially identifying 0.34% of unaffected pregnancies as screen positive. About 1 in 90 screen positive pregnancies will be affected. CONCLUSIONS: It is possible to screen for SLOS as an add on to existing 2nd trimester maternal serum screening, if uE3 is already being measured. A large, prospective trial is necessary to determine whether diagnostic testing can be performed in maternal urine or serum rather than amniotic fluid. PMID- 11943801 TI - Molecular changes of preclinical scrapie can be detected by infrared spectroscopy. AB - Infrared (IR) microspectroscopy was used to detect disease-associated molecular changes spatially resolved in cryosections of scrapie-infected tissue of the CNS. The results show that IR spectra can be used for the discrimination between normal and 263K scrapie-infected hamster nervous tissue not only in the terminal stage of the disease but also in early clinical and even in the preclinical stage at 90 d after oral infection. The nuclei of the cranial nerves located in the medulla oblongata were especially well suited for an early detection of the diseased state by IR microspectroscopy. The most prominent molecular changes indicated by the IR spectra were located between 1300 and 1000 cm(-1), a region that contains contributions primarily from carbohydrates and the phosphate backbones of nucleic acids but also from membrane constituents. PMID- 11943802 TI - Opioid peptides inhibit excitatory but not inhibitory synaptic transmission in the rat dorsal motor nucleus of the vagus. AB - Opioid peptides produce gastrointestinal inhibition and increase feeding when applied to the brainstem. The present studies were designed to determine the actions of opioid peptides on synaptic transmission within the dorsal motor nucleus of the vagus (DMV) and the localization of mu-opioid receptors. Whole cell recordings were made from identified gastrointestinal-projecting DMV neurons in thin brainstem slices of the rat. Electrical stimulation of the nucleus of the tractus solitarius evoked EPSCs and IPSCs. In all neurons tested, methionine (Met)-enkephalin (0.003-30 microm) inhibited the peak amplitude of the EPSCs. The effect was prevented by naloxone (1 microm) as well as by naloxonazine (0.2 microm). An increase in the ratio of the evoked paired pulses indicated that the inhibition was attributable to actions at presynaptic receptors. This presynaptic inhibitory action was mimicked by [d-Ala(2), N-Me-Phe(4), Gly(5)-ol]-enkephalin (0.1 microm) and the analgesic dipeptide kyotorphin (10 microm) but not by cyclic[d-Pen(2), d-Pen(5)]-enkephalin (1 microm) and trans-3,4-dichloro-N-methyl N-[2-(1-pyrrolidinyl)-cyclohexyl]benzeneacetamide methanesulfonate (1 microm). In contrast, the amplitude of evoked IPSCs was not altered either by Met-enkephalin or by any of the opioid receptor-selective agonists. Immunohistochemical studies revealed that nerve terminals apposing DMV neurons showed immunoreactivity to mu opioid receptors colocalized with glutamate immunoreactivity but not glutamic acid decarboxylase immunoreactivity. These results suggest that within the DMV, mu-opioid receptors are present on the nerve terminals of excitatory but not inhibitory inputs to GI motoneurons. Such specificity may imply that the central inhibitory action of opioid peptides on gastrointestinal function targets selected pathways. PMID- 11943803 TI - A developmental switch of AMPA receptor subunits in neocortical pyramidal neurons. AB - AMPA receptors mediate most of the fast excitatory neurotransmission in the brain, and those lacking the glutamate receptor 2 (GluR2) subunit are Ca(2+) permeable and expressed in cortical structures primarily by inhibitory interneurons. Here we report that synaptic AMPA receptors of excitatory layer 5 pyramidal neurons in the rat neocortex are deficient in GluR2 in early development, approximately before postnatal day 16, as evidenced by their inwardly rectifying current-voltage relationship, blockade of AMPA receptor mediated EPSCs by external and internal polyamines, permeability to Ca(2+), and GluR2 immunoreactivity. Overall, these results indicate that neocortical pyramidal neurons undergo a developmental switch in the Ca(2+) permeability of their AMPA receptors through an alteration of their subunit composition. This has important implications for plasticity and neurotoxicity. PMID- 11943804 TI - The neuronal adhesion protein TAG-1 is expressed by Schwann cells and oligodendrocytes and is localized to the juxtaparanodal region of myelinated fibers. AB - The neural cell adhesion molecule TAG-1, which is a glycosylphosphatidylinositol linked member of the Ig superfamily, is expressed by various neuronal populations in the developing CNS and PNS. We demonstrate here that Schwann cells and oligodendrocytes also express TAG-1. In the PNS, TAG-1 is detected in ensheathing Schwann cells early postnatally and is maintained throughout adulthood. In mature myelinated fibers of the CNS and PNS, TAG-1 is localized to the juxtaparanodal region. The CNS of the UDP-galactose ceramide galactosyl transferase(-/-) (CGT(-/ )) mouse mutants, which do not synthesize the abundant galactolipids of myelin, display severely disrupted axoglial interactions at the paranodal region. In contrast, axoglial interactions in the PNS of these mutants are less affected. Interestingly, TAG-1 localization is completely undetected in myelinated fibers of the CNS. In the PNS of these mutants, TAG-1 abnormally localizes in the paranodal region. These data raise the intriguing possibility that TAG-1 localization in the juxtaparanodal area mediates, or at least requires, the axoglial contact normally displayed in this region. The abnormal localization of TAG-1 in the CGT mutants might contribute to the disrupted axoglial interactions observed in these animals. PMID- 11943805 TI - Target depletion of distinct tumor necrosis factor receptor subtypes reveals hippocampal neuron death and survival through different signal transduction pathways. AB - Tumor necrosis factor receptor-I (TNFRI) and TNFRII are two TNFR subtypes in the immune system, but their roles in the brain remain unclear. Here we present a novel interaction between TNFR subtypes and TNF-alpha in the brain. Our studies on target-depleted TNFR in mice show that TNF-alpha has little effect on hippocampal neurons in which TNFRI, containing an "intracellular death domain," is absent (TNFRI -/-), whereas neurons from TNFRII knock-out mice are vulnerable to TNF-alpha even at low doses. Moreover, little nuclear factor-kappaB (NF kappaB) translocation is induced by TNF-alpha in neurons of TNFRI -/-, whereas NF kappaB subunit p65 is still translocated from the cytoplasm into the nucleus in neurons from wild-type and TNFRII -/- mice. Furthermore, p38 mitogen-activated protein (MAP) kinase activity is upregulated in neurons from both wild-type and TNFRI -/-, but no alteration of p38 MAP kinase was found in neurons from TNFRII. Results from overexpression of TNF receptors further support the above findings. NT2 neuronal-like cells transiently transfected with TNFRI are very sensitive to TNF-alpha, whereas TNF-alpha is not toxic and even seems to be trophic to the cells with TNFRII overexpression. Last, our radioligand-binding experiments demonstrate that TNF-alpha binds TNFRI with high affinity (K(d) of 0.6 nm), whereas TNFRII shows lower binding affinity (K(d) of 1.14 nm) to TNF-alpha in NT2 transfected cells. Together, these studies reveal novel neuronal responses of TNF alpha in mediating consequences of TNF receptor activation differently. Subsequent neuronal death or survival may ultimately depend on a particular subtype of TNF receptor that is predominately expressed in neurons of the brain during neural development or with neurological diseases. PMID- 11943806 TI - Odorant receptor expression defines functional units in the mouse olfactory system. AB - Odorant receptors (ORs) mediate the interaction of odorous compounds with olfactory sensory neurons (OSNs) and influence the guidance of OSN axons to synaptic targets in the olfactory bulb (OB). OSNs expressing the same OR send convergent axonal projections to defined glomeruli in the OB and are thought to share the same odorant response properties. This expectation of functional similarity has not been tested experimentally, because it has not been possible to determine reproducibly the response properties of OSNs that express defined ORs. Here, we applied calcium imaging to characterize the odorant response properties of single neurons from gene-targeted mice in which the green fluorescent protein is coexpressed with a particular OR. We show that the odorants acetophenone and benzaldehyde are agonists for the M71 OR and that M71 expressing neurons are functionally similar in their response properties across concentration. Replacing the M71 coding sequence with that of the rat I7 OR changes the stimulus response profiles of this genetically defined OSN population and concomitantly results in the formation of novel glomeruli in the OB. We further show that the mouse I7 OR imparts a particular response profile to OSNs regardless of the epithelial zone of expression. Our data provide evidence that ORs determine both odorant specificity and axonal convergence and thus direct functionally similar afferents to form particular glomeruli. They confirm and extend the notion that OR expression provides a molecular basis for the formation and arrangement of glomerular functional units. PMID- 11943807 TI - Regulation of GluR1 by the A-kinase anchoring protein 79 (AKAP79) signaling complex shares properties with long-term depression. AB - Second messengers regulate synaptic plasticity by influencing the balance between kinase and phosphatase activity. One target of this balance is the phosphorylation state of the AMPA receptor glutamate receptor 1 (GluR1) subunit. Hippocampal long-term depression (LTD) is a calcium-dependent downregulation of synaptic AMPA receptor currents associated with dephosphorylation of Ser845, a cAMP-dependent protein kinase (PKA) site on GluR1. Recruitment of kinases and phosphatases to the AMPA receptor might enable modulation of AMPA receptor function. The neuronal A-kinase anchoring protein AKAP79/150 interacts with PKA and the calcium-dependent protein phosphatase PP2B and is linked to the AMPA receptor GluR1 subunit by synapse-associated protein 97 (SAP97), a membrane associated guanylate kinase family protein. Here we demonstrate that AKAP79 not only promotes basal phosphorylation of Ser845 but also confers a calcium- and PP2B-mediated downregulation to GluR1 receptor currents. This AKAP79-dependent downregulation is contingent on the local presence of PKA, Ser845 of GluR1, and a PDZ (postsynaptic density 95/Discs large/zona occludens 1)-domain interaction between GluR1 and SAP97, all of which support basal phosphorylation of the receptor. These findings suggest that the AKAP79 signaling complex is sufficient to couple intracellular calcium levels to the PKA phosphorylation state of GluR1. Thus, the integration of intracellular signals relevant for LTD may be transduced to GluR1 by the AKAP79 signaling complex. PMID- 11943808 TI - The cytokine network of Wallerian degeneration: tumor necrosis factor-alpha, interleukin-1alpha, and interleukin-1beta. AB - Wallerian degeneration (WD) is the inflammatory response of the nervous system to axonal injury, primarily attributable to the production of cytokines, the mediator molecules of inflammation. We presently document the involvement of the inflammatory cytokines TNFalpha, interleukin (IL)-1alpha, and IL-1beta in peripheral nerve (PNS) injury in C57/BL/6NHSD (C57/BL) mice that display the normal rapid progression of WD (rapid-WD) and C57/BL/6-WLD/OLA/NHSD mice that display abnormal slow progression of WD (slow-WD). TNFalpha and IL-1alpha mRNAs were expressed, whereas TNFalpha but not IL-1alpha protein was synthesized in intact PNS of C57/BL mice. TNFalpha and IL-1alpha protein synthesis and secretion were rapidly upregulated during rapid-WD in Schwann cells. IL-1beta mRNA expression and protein synthesis and secretion were induced sequentially in Schwann cells with a delay after injury. Thereafter, recruited macrophages contributed to the production of TNFalpha, IL-1alpha, and IL-1beta, which in turn augmented myelin phagocytosis by macrophages. Observations suggest that TNFalpha and IL-1alpha are the first cytokines with protein production that is upregulated during rapid-WD. TNFalpha and IL-1alpha may initiate, therefore, molecular and cellular events in rapid-WD (e.g., the production of additional cytokines and NGF). TNFalpha, IL-1alpha, and IL-1beta may further regulate, indirectly, macrophage recruitment, myelin removal, regeneration, and neuropathic pain. In contrast to rapid-WD, the production of TNFalpha, IL-1alpha, and IL-1beta protein was deficient in slow-WD, although their mRNAs were expressed. mRNA expression and protein production of TNFalpha, IL-1alpha, and IL-1beta were differentially regulated during rapid-WD and slow-WD, suggesting that mRNA expression, by itself, is no indication of the functional involvement of cytokines in WD. PMID- 11943809 TI - ADP and AMP induce interleukin-1beta release from microglial cells through activation of ATP-primed P2X7 receptor channels. AB - P2X(7) is a subtype of ATP-gated channels that is highly expressed in astrocytes, microglia, and other immune cells. Activation of P2X(7) purinoceptors by ATP or 3'-O-(4-benzoyl)-benzoyl ATP (BzATP) induces the formation of cytolytic pores and provokes release of interleukin-1beta from immune cells. We investigated the actions of other endogenous nucleotides on recombinant and microglial P2X(7) receptors using electrophysiology, fluorescence imaging, and interleukin-1beta release measurement. We found that initial application of ADP or AMP to Xenopus oocytes expressing P2X(7) receptors was ineffective. However, when ADP and AMP, but not UTP or adenosine, were applied after a brief exposure to ATP or BzATP, they activated P2X(7) receptors in a dose-dependent manner. Moreover, responses to ADP and AMP were also elicited after exposure to low concentrations of ATP and were recorded several minutes after removal of ATP from the extracellular medium. Whole-cell recordings from mouse microglial cells showed that significant responses to ADP and AMP were elicited only after ATP application. YO-PRO-1 dye uptake imaging revealed that, unlike ATP, prolonged application of ADP or AMP did not cause an opening of large cytolytic pores in mouse microglial cells. Finally, ADP and AMP stimulated the release of interleukin-1beta from ATP-primed mouse and human microglial cells. We conclude that selective sensitization of P2X(7) receptors to ADP and AMP requires priming with ATP. This novel property of P2X(7) leads to activation by ATP metabolites and proinflammatory cytokine release from microglia without cytotoxicity. PMID- 11943810 TI - NMDA receptor antagonists disinhibit rat posterior cingulate and retrosplenial cortices: a potential mechanism of neurotoxicity. AB - NMDA receptor antagonists produce region-specific neurodegeneration by an undetermined mechanism, but one proposed mechanism involves disinhibition. In certain areas of the brain, NMDA receptors mediate excitatory drive onto inhibitory interneurons. Thus, NMDA receptor/channel antagonists may reduce inhibition (i.e., produce "disinhibition"). If a sufficient level of disinhibition is produced, enhanced vulnerability to excitotoxicity may result. Furthermore, if there are region-specific differences in NMDA antagonist-induced disinhibition, this could underlie region-specific NMDA antagonist-induced neurotoxicity. In the present study, we tested this hypothesis by exposing rat brain slices to the NMDA receptor antagonist dizocilpine maleate (MK-801) and measuring MK-801-induced disinhibition in areas of higher and lower vulnerability to neurodegeneration [posterior cingulate/retrosplenial cortices (PCC/RSC) and parietal cortex, respectively]. Using whole-cell patch-clamp techniques, bicuculline-sensitive GABA(A) receptor-mediated IPSCs were measured in biocytin labeled pyramidal neurons in the PCC/RSC and parietal cortex. In the PCC/RSC, bath-applied MK-801 (10-40 microm) produced disinhibition, shown as a concentration-dependent decrease in spontaneous IPSC frequency and amplitude; MK 801 (40 microm) also reduced evoked IPSC amplitudes. In parietal cortex, MK-801 produced significantly less disinhibition. To determine whether disinhibition is caused by presynaptic or postsynaptic mechanisms, we tested the effects of MK-801 (40 microm) against miniature IPSC (mIPSC) frequency and amplitude in tetrodotoxin (TTX; 0.5 microm)-treated slices and found that MK-801 did not alter mIPSC frequency or amplitude. Taken together, these results suggest that NMDA receptors regulate activity of inhibitory interneurons and, consequently, GABA release in certain cortical areas. This region-specific reduction in inhibitory input to pyramidal cells could underlie the region-specific neurotoxicity of NMDA antagonists. PMID- 11943811 TI - Vitamin E but not 17beta-estradiol protects against vascular toxicity induced by beta-amyloid wild type and the Dutch amyloid variant. AB - Amyloid beta-peptide (Abeta) fibril deposition on cerebral vessels produces cerebral amyloid angiopathy that appears in the majority of Alzheimer's disease patients. An early onset of a cerebral amyloid angiopathy variant called hereditary cerebral hemorrhage with amyloidosis of the Dutch type is caused by a point mutation in Abeta yielding Abeta(Glu22-->Gln). The present study addresses the effect of amyloid fibrils from both wild-type and mutated Abeta on vascular cells, as well as the putative protective role of antioxidants on amyloid angiopathy. For this purpose, we studied the cytotoxicity induced by Abeta(1-40 Glu22-->Gln) and Abeta(1-40 wild-type) fibrils on human venule endothelial cells and rat aorta smooth muscle cells. We observed that Abeta(Glu22-->Gln) fibrils are more toxic for vascular cells than the wild-type fibrils. We also evaluated the cytotoxicity of Abeta fibrils bound with acetylcholinesterase (AChE), a common component of amyloid deposits. Abeta(1-40 wild-type)-AChE fibrillar complexes, similar to neuronal cells, resulted in an increased toxicity on vascular cells. Previous reports showing that antioxidants are able to reduce the toxicity of Abeta fibrils on neuronal cells prompted us to test the effect of vitamin E, vitamin C, and 17beta-estradiol on vascular damage induced by Abeta(wild-type) and Abeta(Glu22-->Gln). Our data indicate that vitamin E attenuated significantly the Abeta-mediated cytotoxicity on vascular cells, although 17beta-estradiol and vitamin C failed to inhibit the cytotoxicity induced by Abeta fibrils. PMID- 11943812 TI - A role for alpha-synuclein in the regulation of dopamine biosynthesis. AB - The alpha-synuclein gene is implicated in the pathogenesis of Parkinson's disease. Although alpha-synuclein function is uncertain, the protein has homology to the chaperone molecule 14-3-3. In addition, alpha-synuclein can bind to 14-3 3, and both alpha-synuclein and 14-3-3 bind to many of the same proteins. Because 14-3-3 binds to and activates tyrosine hydroxylase, the rate-limiting enzyme in dopamine (DA) biosynthesis, we explored whether alpha-synuclein also bound to tyrosine hydroxylase and influenced its activity. Immunoprecipitation revealed an interaction between alpha-synuclein and tyrosine hydroxylase in brain homogenates and MN9D dopaminergic cells. Colocalization of alpha-synuclein with tyrosine hydroxylase was confirmed by immunoelectron microscopy. To explore the consequences of the interaction, we measured the effect of recombinant alpha synuclein on tyrosine hydroxylase activity in a cell-free system and observed a dose-dependent inhibition of tyrosine hydroxylase by alpha-synuclein. To measure the impact of alpha-synuclein on tyrosine hydroxylase in dopaminergic cells, we stably transfected MN9D cells with wild-type or A53T mutant alpha-synuclein. Overexpression of wild-type or A53T mutant alpha-synuclein did not significantly alter tyrosine hydroxylase protein levels in our stably transfected cells. However, overexpressing cell lines had significantly reduced tyrosine hydroxylase activity and a corresponding reduction in dopamine synthesis. The reduction in cellular dopamine levels was not caused by increased dopamine catabolism or dopamine efflux. These data suggest that alpha-synuclein plays a role in the regulation of dopamine biosynthesis, acting to reduce the activity of tyrosine hydroxylase. If so, a loss of soluble alpha-synuclein, by reduced expression or aggregation, could increase dopamine synthesis with an accompanying increase in reactive dopamine metabolites. PMID- 11943813 TI - A role for phosphorylation in the maintenance of resurgent sodium current in cerebellar purkinje neurons. AB - Cerebellar Purkinje neurons express voltage-gated, tetrodotoxin (TTX)-sensitive sodium channels that not only open and inactivate rapidly during depolarization but also reopen during repolarization, carrying an unusual "resurgent" sodium current. Expression of Na(V)1.6 alpha subunits appears necessary but not sufficient to generate this component of current; Purkinje cells without Na(V)1.6 lack resurgent current, but resurgent current is absent from many other Na(V)1.6 expressing neurons. These observations raise the question of how modulation or modification of the Na(V)1.6 subunit may lead to production of resurgent current. Previous studies have suggested that sodium channels of Purkinje neurons are subject to a rapid, voltage-dependent, open channel block by an endogenous particle whose unbinding allows resurgent current to flow. To investigate the nature of this block, we recorded TTX-sensitive sodium currents in outside-out patches from Purkinje cells acutely isolated from mice. In all patches, step depolarizations evoked transient current, and step repolarizations evoked resurgent current. The amplitudes of the transient and resurgent currents were highly correlated across patches (R(2) > 0.99), suggesting that the blocking agent is closely associated with the channel. Intracellular protease eliminated fast inactivation, indicating that the blocking element, like the fast inactivation gate, may be proteinaceous. Intracellular application of alkaline phosphatase abolished resurgent current and significantly slowed inactivation of transient current. The phosphatase inhibitor vanadate reduced these effects. Together, the results suggest that constitutive phosphorylation of the sodium channel complex of Purkinje neurons is necessary to maintain a functional blocking element and produce resurgent sodium current. PMID- 11943814 TI - Remodeling of hippocampal synaptic networks by a brief anoxia-hypoglycemia. AB - Cerebral ischemia is a major cause of brain dysfunction. Using a model of delayed death induced by a brief, transient oxygen and glucose deprivation, we studied here how this affected the structural organization of hippocampal synaptic networks. We report that brief anoxic-hypoglycemic episodes rapidly modified the structure of synapses. This was characterized, at the electron microscopic level, by a transient increase in the proportion of perforated synapses, followed after 2 hr by an increase in images of multiple synapse boutons. These changes were considerable because 10-20% of all synapses were affected. This structural remodeling was correlated by three kinds of modifications observed using two photon confocal microscopy: the growth of filopodia, occurring shortly (5-20 min) after anoxia-hypoglycemia, enlargements of existing spines, and formation of new spines, both seen mainly 20-60 min after the insult. All of these structural changes were calcium and NMDA receptor dependent and thus reproduced, to a larger scale, those associated with synaptic plasticity. Concomitantly and related to the severity of anoxia-hypoglycemia, we could also observe spine loss and images of spine, dendrite, or presynaptic terminal swellings that evolved up to membrane disruption. These changes were also calcium dependent and reduced by NMDA receptor antagonists. Thus, short anoxic-hypoglycemic episodes, through NMDA receptor activation and calcium influx, resulted in a profound structural remodeling of synaptic networks, through growth, formation, and elimination of spines and synapses. PMID- 11943815 TI - Ectopic expression in the giant fiber system of Drosophila reveals distinct roles for roundabout (Robo), Robo2, and Robo3 in dendritic guidance and synaptic connectivity. AB - The Roundabout (Robo) receptors have been intensively studied for their role in regulating axon guidance in the embryonic nervous system, whereas a role in dendritic guidance has not been explored. In the adult giant fiber system of Drosophila, we have revealed that ectopic Robo expression can regulate the growth and guidance of specific motor neuron dendrites, whereas Robo2 and Robo3 have no effect. We also show that the effect of Robo on dendritic guidance can be suppressed by Commissureless coexpression. Although we confirmed a role for all three Robo receptors in giant fiber axon guidance, the strong axon guidance alterations caused by overexpression of Robo2 or Robo3 have no effect on synaptic connectivity. In contrast, Robo overexpression in the giant fiber seems to directly interfere with synaptic function. We conclude that axon guidance, dendritic guidance, and synaptogenesis are separable processes and that the different Robo family members affect them distinctly. PMID- 11943816 TI - Use-dependent modulation of inhibitory capacity in the feline lumbar spinal cord. AB - The ability to perform stepping and standing can be reacquired after complete thoracic spinal cord transection in adult cats with appropriate, repetitive training. We now compare GAD(67)A levels in the spinal cord of cats that were trained to step or stand. We confirmed that a complete spinal cord transection at approximately T12 increases glutamic acid decarboxylase (GAD)(67) in both the dorsal and ventral horns of L5-L7. We now show that step training decreases these levels toward control. Kinematic analyses show that this downward modulation is correlated inversely with stepping ability. Compared with intact cats, spinal cord-transected cats had increased punctate GAD(67) immunoreactivity around neurons in lamina IX at cord segments L5-L7. Compared with spinal nontrained cats, those trained to stand on both hindlimbs had more GAD(67) puncta bilaterally in a subset of lamina IX neurons. In cats trained to stand unilaterally, this elevated staining pattern was limited to the trained side and extended for at least 4 mm in the L6 and L7 segments. The location of this asymmetric GAD(67) staining corresponded to the motor columns of primary knee flexors, which are minimally active during standing, perhaps because of extensor activated inhibitory interneuron projections. The responsiveness to only a few days of motor training, as well as the GABA-synthesizing potential in the spinal cord, persists for at least 25 months after the spinal cord injury. This modulation is specific to the motor task that is performed repetitively and is closely linked to the ability of the animal to perform a specific motor task. PMID- 11943817 TI - The critical role of basement membrane-independent laminin gamma 1 chain during axon regeneration in the CNS. AB - We have addressed the question of whether a family of axon growth-promoting molecules known as the laminins may play a role during axon regeneration in the CNS. A narrow sickle-shaped region containing a basal lamina-independent form of laminin exists in and around the cell bodies and proximal portion of the apical dendrites of CA3 pyramidal neurons of the postnatal hippocampus. To understand the possible function of laminin in axon regeneration within this pathway, we have manipulated laminin synthesis at the mRNA level in a slice culture model of the lesioned mossy system. In this model early postnatal mossy fibers severed near the hilus can regenerate across the lesion and elongate rapidly within strata lucidum and pyramidale. In slice cultures of the postnatal day 4 hippocampus, 2 d before lesion and then continuing for 1-5 d after lesion, translation of the gamma1 chain product of laminin was reduced by using antisense oligodeoxyribonucleotides and DNA enzymes. In the setting of the lesioned organotypic hippocampal slice, astroglial repair of the lesion and overall glial patterning were unperturbed by the antisense or DNA enzyme treatments. However, unlike controls, in the treated, lesioned slices the vast majority of regenerating mossy fibers could not cross the lesion site; those that did were very much shorter than usual, and they took a meandering course. In a recovery experiment in which the DNA enzyme or antisense oligos were washed away, laminin immunoreactivity returned and mossy fiber regeneration resumed. These results demonstrate the critical role of laminin(s) in an axon regeneration model of the CNS. PMID- 11943818 TI - Dividing precursor cells of the embryonic cortical ventricular zone have morphological and molecular characteristics of radial glia. AB - The embryonic ventricular zone (VZ) of the cerebral cortex contains migrating neurons, radial glial cells, and a large population of cycling progenitor cells that generate newborn neurons. The latter two cell classes have been assumed for some time to be distinct in both function and anatomy, but the cellular anatomy of the progenitor cell type has remained poorly defined. Several recent reports have raised doubts about the distinction between radial glial and precursor cells by demonstrating that radial glial cells are themselves neuronal progenitor cells (Malatesta et al., 2000; Hartfuss et al., 2001; Miyata et al., 2001; Noctor et al., 2001). This discovery raises the possibility that radial glia and the population of VZ progenitor cells may be one anatomical and functional cell class. Such a hypothesis predicts that throughout neurogenesis almost all mitotically active VZ cells and a substantial percentage of VZ cells overall are radial glia. We have therefore used various anatomical, immunohistochemical, and electrophysiological techniques to test these predictions. Our data demonstrate that the majority of VZ cells, and nearly all mitotically active VZ cells during neurogenesis, both have radial glial morphology and express radial glial markers. In addition, intracellular dye filling of electrophysiologically characterized progenitor cells in the VZ demonstrates that these cells have the morphology of radial glia. Because the vast majority cycling cells in the cortical VZ have characteristics of radial glia, the radial glial precursor cell may be responsible for both the production of newborn neurons and the guidance of daughter neurons to their destinations in the developing cortex. PMID- 11943819 TI - Prolonged seizures increase proliferating neuroblasts in the adult rat subventricular zone-olfactory bulb pathway. AB - Neuronal precursors in the adult rodent forebrain subventricular zone (SVZ) proliferate, migrate to the olfactory bulb in a restricted pathway known as the rostral migratory stream (RMS), and differentiate into neurons. The effects of injury on this neurogenic region of the mature brain are poorly understood. To determine whether seizure-induced injury modulates SVZ neurogenesis, we induced status epilepticus (SE) in adult rats by systemic chemoconvulsant administration and examined patterns of neuronal precursor proliferation and migration in the SVZ-olfactory bulb pathway. Within 1-2 weeks after pilocarpine-induced SE, bromodeoxyuridine (BrdU) labeling and Nissl staining increased in the rostral forebrain SVZ. These changes were associated with an increase in cells expressing antigenic markers of SVZ neuroblasts 2-3 weeks after prolonged seizures. At these same time points the RMS expanded and contained more proliferating cells and immature neurons. BrdU labeling and stereotactic injections of retroviral reporters into the SVZ showed that prolonged seizures also increased neuroblast migration to the olfactory bulb and induced a portion of the neuronal precursors to exit the RMS prematurely. These findings indicate that SE expands the SVZ neuroblast population and alters neuronal precursor migration in the adult rat forebrain. Identification of the mechanisms underlying the response of neural progenitors to seizure-induced injury may help to advance brain regenerative therapies by using either transplanted or endogenous neural precursor cells. PMID- 11943820 TI - The timing of response onset and offset in macaque visual neurons. AB - We used fast, pseudorandom temporal sequences of preferred and antipreferred stimuli to drive neuronal firing rates rapidly between minimal and maximal across the visual system. Stimuli were tailored to the preferences of cells recorded in the lateral geniculate nucleus (magnocellular and parvocellular), primary visual cortex (simple and complex), and the extrastriate motion area MT. We found that cells took longer to turn on (to increase their firing rate) than to turn off (to reduce their rate). The latency difference (onset minus offset) varied from several to tens of milliseconds across cell type and stimulus class and was correlated with spontaneous or driven firing rates for most cell classes. The delay for response onset depended on the nature of the stimulus present before the preferred stimulus appeared, and may result from persistent inhibition caused by antipreferred stimuli or from suppression that followed the offset of the preferred stimulus. The onset delay showed three distinct types of dependence on the temporal sequence of stimuli across classes of cells, implying that suppression may accumulate or wear off with time. Onset latency is generally longer, can be more variable, and has marked stimulus dependence compared with offset latency. This suggests an important role for offset latency in assessing the speed of information transmission in the visual system and raises the possibility that signal offsets provide a timing reference for visual processing. We discuss the origin of the delay in onset latency compared with offset latency and consider how it may limit the utility of certain feedforward circuits. PMID- 11943821 TI - Does anticipation of pain affect cortical nociceptive systems? AB - Anticipation of pain is a complex state that may influence the perception of subsequent noxious stimuli. We used functional magnetic resonance imaging (fMRI) to study changes of activity of cortical nociceptive networks in healthy volunteers while they expected the somatosensory stimulation of one foot, which might be painful (subcutaneous injection of ascorbic acid) or not. Subjects had no previous experience of the noxious stimulus. Mean fMRI signal intensity increased over baseline values during anticipation and during actual stimulation in the putative foot representation area of the contralateral primary somatosensory cortex (SI). Mean fMRI signals decreased during anticipation in other portions of the contralateral and ipsilateral SI, as well as in the anteroventral cingulate cortex. The activity of cortical clusters whose signal time courses showed positive or negative correlations with the individual psychophysical pain intensity curve was also significantly affected during the waiting period. Positively correlated clusters were found in the contralateral SI and bilaterally in the anterior cingulate, anterior insula, and medial prefrontal cortex. Negatively correlated clusters were found in the anteroventral cingulate bilaterally. In all of these areas, changes during anticipation were of the same sign as those observed during pain but less intense ( approximately 30-40% as large as peak changes during actual noxious stimulation). These results provide evidence for top-down mechanisms, triggered by anticipation, modulating cortical systems involved in sensory and affective components of pain even in the absence of actual noxious input and suggest that the activity of cortical nociceptive networks may be directly influenced by cognitive factors. PMID- 11943823 TI - Dendro-dendritic interactions between motion-sensitive large-field neurons in the fly. AB - For visual course control, flies rely on a set of motion-sensitive neurons called lobula plate tangential cells (LPTCs). Among these cells, the so-called CH (centrifugal horizontal) cells shape by their inhibitory action the receptive field properties of other LPTCs called FD (figure detection) cells specialized for figure-ground discrimination based on relative motion. Studying the ipsilateral input circuitry of CH cells by means of dual-electrode and combined electrical-optical recordings, we find that CH cells receive graded input from HS (large-field horizontal system) cells via dendro-dendritic electrical synapses. This particular wiring scheme leads to a spatial blur of the motion image on the CH cell dendrite, and, after inhibiting FD cells, to an enhancement of motion contrast. This could be crucial for enabling FD cells to discriminate object from self motion. PMID- 11943822 TI - Domoic acid lesions in nucleus of the solitary tract: time-dependent recovery of hypoxic ventilatory response and peripheral afferent axonal plasticity. AB - The nucleus of the solitary tract (NTS) plays a pivotal role in the ventilatory response to hypoxia (HVR). However, the effects of excitotoxic lesions and the potential for functional recovery and plasticity remain unknown. Domoic acid (DA) or vehicle were bilaterally injected within the NTS of adult male Sprague Dawley rats. HVR (10% O(2)) and anatomical changes were assessed at 5-90 d after surgery. DA induced dose-dependent HVR attenuations ( approximately 70% at peak effect) that exhibited saturation at concentrations of 0.75-1.0 mm. However, although sodium cyanide-induced ventilatory responses were virtually abolished, DA did not modify baroreceptor gain. Consistent with ventilatory reductions, NTS neurons showed a significant degeneration 3 d after DA injection. In addition, the projection fields and the density of vagal afferent terminals to the NTS, and the motor neurons in the dorsal motor nucleus of the vagus were substantially reduced at 15 d. At 30 d, no functional or neural recovery were apparent. However, at day 60, the reduction in HVR was only approximately 40% of control, and at 90 d, HVR returned to control levels, paralleling regeneration of vagal afferent terminals within the NTS. The regeneration was particularly prominent in the commissural and dorsomedial subnuclei in the absence of cellular recovery. Thus, the integrity of the NTS is critical for HVR, spontaneous HVR recovery occurs after excitotoxic lesions in the NTS, and vagal-glossopharyngeal terminal sprouting in the NTS may underlie the anatomical substrate for such spontaneous functional recovery. The adult brainstem/NTS has self-repairing capabilities and will compensate for functional losses after structural damage by rewiring of its neural circuitry. PMID- 11943824 TI - Cholinergic changes in the APP23 transgenic mouse model of cerebral amyloidosis. AB - Alzheimer's Disease (AD) is a neurodegenerative disorder that is characterized by extracellular deposits of amyloid-beta peptide (Abeta) and a severe depletion of the cholinergic system, although the relationship between these two events is poorly understood. In the neocortex, there is a loss of cholinergic fibers and receptors and a decrease of both choline acetyltransferase (ChAT) and acetylcholinesterase enzyme activities. The nucleus basalis of Meynert (NBM), which provides the major cholinergic input to the neocortex, undergoes profound neuron loss in AD. In the present study, we have examined the cholinergic alterations in amyloid precursor protein transgenic mice (APP23), a mouse model of cerebral beta-amyloidosis. In aged APP23 mice, our results reveal modest decreases in cortical cholinergic enzyme activity compared with age-matched wild type mice. Total cholinergic fiber length was more severely affected, with 29 and 35% decreases in the neocortex of aged APP23 mice compared with age-matched wild type mice and young transgenic mice, respectively. However, there was no loss of cholinergic basal forebrain neurons in these aged APP23 mice, suggesting that the cortical cholinergic deficit in APP23 mice is locally induced by the deposition of amyloid and is not caused by a loss of cholinergic basal forebrain neurons. To study the impact of cholinergic basal forebrain degeneration on cortical amyloid deposition, we performed unilateral NBM lesions in adult APP23 mice. Three to 8 months after lesioning, a 38% reduction in ChAT activity and significant cholinergic fiber loss were observed in the ipsilateral frontal cortex. There was a 19% decrease in Abeta levels of the ipsilateral compared with contralateral frontal cortex with no change in the ratio of Abeta40 to Abeta42. We conclude that the severe cholinergic deficit in AD is caused by both the loss of cholinergic basal forebrain neurons and locally by cerebral amyloidosis in the neocortex. Moreover, our results suggest that disruption of the basal cholinergic forebrain system does not promote cerebral amyloidosis in APP23 transgenic mice. PMID- 11943825 TI - The rate of intravenous cocaine administration determines susceptibility to sensitization. AB - The potential for addiction is thought to be greatest when drugs of abuse reach the brain rapidly, because this produces intense subjective pleasurable effects. However, the ability of drugs to induce forms of cellular plasticity related to behavioral sensitization may also contribute to addiction. Therefore, we studied the influence of rate of intravenous cocaine delivery on its ability to induce psychomotor sensitization. In one experiment, rotational behavior in rats with a unilateral 6-hydroxydopamine lesion was used as an index of psychomotor activation, and in a second experiment, locomotor activity in neurologically intact rats was used. Rapid (5-16 sec) intravenous infusions of cocaine induced robust psychomotor sensitization at all doses tested (0.5-2.0 mg/kg). Treatments given over 25 sec failed to induce sensitization at all doses tested. Treatments given over 50 or 100 sec induced sensitization only at the highest dose tested. Thus, the rate of intravenous cocaine delivery has profound effects on the ability of cocaine to induce psychomotor sensitization. This suggests that the temporal dynamics of drug delivery to the brain is a critical factor in the ability of cocaine to induce forms of neuronal plasticity that may contribute to addiction. PMID- 11943826 TI - Brain-derived neurotrophic factor produces antidepressant effects in behavioral models of depression. AB - Previous studies demonstrated that antidepressant treatment increases the expression of brain-derived neurotrophic factor (BDNF) in rat hippocampus. The present study was conducted to test the hypothesis that BDNF in the hippocampus produces an antidepressant effect in behavioral models of depression, the learned helplessness (LH) and forced swim test (FST) paradigms. A single bilateral infusion of BDNF into the dentate gyrus of hippocampus produced an antidepressant effect in both the LH and FST that was comparable in magnitude with repeated systemic administration of a chemical antidepressant. These effects were observed as early as 3 d after a single infusion of BDNF and lasted for at least 10 d. Similar effects were observed with neurotrophin-3 (NT-3) but not nerve growth factor. Infusions of BDNF and NT-3 did not influence locomotor activity or passive avoidance. The results provide further support for the hypothesis that BDNF contributes to the therapeutic action of antidepressant treatment. PMID- 11943827 TI - cAMP response element-binding protein is essential for the upregulation of brain derived neurotrophic factor transcription, but not the behavioral or endocrine responses to antidepressant drugs. AB - Antidepressant drugs activate the cAMP signal transduction pathway through a variety of monoamine neurotransmitter receptors. Recently, molecular studies have identified a role for cAMP response element-binding protein (CREB) in the mechanism of action of chronically administered antidepressant drugs. However, the function of CREB in the behavioral and endocrine responses to these drugs has not been thoroughly investigated. We have used CREB-deficient mice to study the effects of two antidepressants, desipramine (DMI) and fluoxetine (FLX), in behavioral, endocrine, and molecular analyses. Behaviorally, CREB-deficient mice and wild-type mice respond similarly to DMI and FLX administration in the forced swim test and tail suspension test. Furthermore, the ability of DMI to suppress an acute corticosterone response after swim stress is maintained in CREB deficient mice. However, upregulation of a molecular target of CREB, BDNF, is abolished in the CREB-deficient mice after chronic administration of DMI. These data are the first to demonstrate that CREB activation is upstream of BDNF mechanistically in response to antidepressant drug treatment. Therefore, although behavioral and endocrine responses to antidepressants may occur by CREB independent mechanisms, CREB is critical to target gene regulation after chronic drug administration, which may contribute to long-term adaptations of the system to antidepressant drug treatment. PMID- 11943828 TI - Psychostimulant-induced behavioral sensitization depends on nicotinic receptor activation. AB - Animal studies have shown that nicotine and psychostimulant drugs (amphetamine and cocaine) share the property of inducing long-lasting behavioral and neurochemical sensitization, which is thought to contribute to their addictive properties. Neuroplasticity subserving learning and memory mechanisms is considered to be involved in psychostimulant-induced sensitization and addiction behavior. Because nicotinic receptors in the brain play a role in the storage of drug-related information underlying reinforcement learning, we evaluated the possibility that activation of central nicotinic receptors may underlie psychostimulant-induced sensitization. Repeated exposure of rats to nicotine profoundly enhanced the psychomotor effects of nicotine and amphetamine 3 weeks after nicotine pretreatment. Moreover, the nicotinic receptor antagonist mecamylamine completely blocked the induction, but not the long-term expression, of behavioral sensitization to amphetamine in amphetamine-pretreated rats. Mecamylamine also prevented the development of cocaine-induced behavioral sensitization. Behavioral sensitization induced by nicotine, amphetamine, or cocaine was associated with an increase in the electrically evoked release of [(3)H]dopamine from nucleus accumbens slices. Coadministration of mecamylamine during pretreatment with nicotine, amphetamine, or cocaine prevented the development of this long-term hyperreactivity of nucleus accumbens dopamine neurons. Similarly, the high-affinity non-alpha7 subtype nicotinic receptor antagonist dihydro-beta-erythroidine prevented the development of amphetamine induced behavioral and neurochemical sensitization. These data indicate that nicotinic receptor activation (by endogenously released acetylcholine) is a common denominator initiating neuroplasticity involved in the development of amphetamine, as well as cocaine-induced sensitization. PMID- 11943829 TI - Midline thalamic region: widespread excitatory input to the entorhinal cortex and amygdala. AB - The midline thalamus has a role in memory formation and has well described projections to multiple limbic sites including the hippocampus, amygdala, and entorhinal cortex. Stimulation of this region evokes excitatory responses in the CA1 region of the hippocampus, but nothing is known about the nature of thalamic influence on other limbic sites such as the entorhinal cortex and the amygdala. In this study we electrically stimulated the midline thalamus in anesthetized rats to determine whether responses could be evoked in the amygdala or entorhinal cortex. In addition we examined the distribution of the responses within the target regions as well as the effect of short interval paired or high-frequency tetanizing stimulation. We found reproducible responses in the entorhinal cortex and the amygdala with a distribution of responses that matched the described synaptic input from the thalamus. In addition, high-frequency stimulation induced a consistent long-term potentiation in the two sites. Paired stimulation resulted in depression of the test response in the amygdala, but a facilitation in the entorhinal cortex. These findings indicate that the midline has a significant monosynaptic excitatory influence in the amygdala and the entorhinal cortex. Combined with the previous work in the hippocampus, this study suggests that the midline thalamus plays a significant role in limbic physiology and may serve to synchronize activity in this system. PMID- 11943830 TI - GABA transporters regulate inhibition in the retina by limiting GABA(C) receptor activation. AB - Inhibition is mediated by two classes of ionotropic receptors in the retina, GABA(A) and GABA(C) receptors. We used the GABA transport blocker NO-711 to examine the role of GABA transporters in shaping synaptic responses mediated by these two receptors in the salamander retinal slice preparation. Focal applications (puffs) of GABA onto GABA(C) receptors on bipolar cells terminals or GABA(A) receptors on ganglion cells elicited currents that were enhanced by NO 711, demonstrating the presence of transporters in the inner plexiform layer (IPL). IPSCs were evoked in bipolar and ganglion cells by puffing kainate into the IPL. NO-711 enhanced the IPSCs only in bipolar cells, suggesting that, when GABA uptake was blocked, the GABA(C) receptors were more strongly activated by spillover transmission than the GABA(A) receptors on ganglion cells. NO-711 enhanced the light-evoked IPSCs mediated by GABA(C) receptors on bipolar cell axon terminals, which resulted in reduced transmission between bipolar and ganglion cells. NO-711 also shifted the intensity-response relationship of the ganglion cell, reducing its sensitivity to light. Surround illumination has been shown by others to produce similar shifts in ganglion cell light sensitivity. Our results show that GABA transporters limit the extent of inhibitory transmission at the inner retina during light-evoked signal processing. PMID- 11943831 TI - Changes in extracellular dopamine induced by morphine and cocaine: crucial control by D2 receptors. AB - An increase of extracellular dopamine (DA) concentration is a major neurobiological substrate of the addictive properties of drugs of abuse. In this article we investigated the contribution of the DA D2 receptor (D2R) in the control of this response. Extracellular DA levels were measured in the striatum of mice lacking D2R expression (D2R-/-) by in vivo microdialysis after administration of the psychostimulant cocaine and the opioid morphine. Interestingly, the increase in extracellular DA induced by both drugs was strikingly higher in D2R-/- than in wild-type littermates. This indicates that D2Rs play a key role in the modulation of DA release in response to drugs of abuse. Furthermore, this observation prompted us to investigate the dopaminergic autoreceptor function in the absence of D2 receptor in D2R-/- mice. Results obtained using complementary microdialysis and voltammetry analyses show that the autoreceptor function regulating DA release is totally abolished in the absence of D2R, despite unchanged DA uptake and basal DA efflux. Finally, we propose that the short isoform D2S receptor of the D2 receptors is the one controlling change in DA release induced by drugs of abuse. Indeed, the neurochemical effects of cocaine and morphine are unchanged in animals with a selective deletion of the long isoform D2L receptor. Thus, deregulated expression of D2R isoforms might be involved in the vulnerability of an individual to drug abuse. PMID- 11943833 TI - Overweight, waist circumference, age, gender, and insulin resistance as risk factors for hyperleptinemia. AB - OBJECTIVE: To evaluate the influence of overweight, waist circumference, age, gender, and insulin resistance as risk factors for hyperleptinemia. RESEARCH METHODS AND PROCEDURES: A cross-sectional study was carried out in a population of 197 subjects: 59 men (21 aged < 60 years and 38 aged > or =60 years) and 138 women (37 aged < 60 years and 101 aged > or =60 years). The groups were stratified by overweight and normal weight. After a 12-hour fasting period, we measured serum leptin and insulin levels with radioimmunoassay methods. We also measured serum glucose and lipid profile. The data were analyzed by means of comparative tests. A variance-stabilizing transformation (natural logarithmic) was used to meet multiple linear regression, analysis of covariance, and logistic regression models. RESULTS: The leptin serum levels were higher and statistically significant in young and older women than they were in men. We observed an interaction between gender and body mass index to explain the difference in leptin levels (p < 0.0001). Our study demonstrated an inverse relationship between leptin with age and high-density lipoprotein cholesterol. In logistic regression analysis, the overweight x gender interaction and waist circumference have a statistically significant influence as independent variables on hyperleptinemia (overweight x gender odds ratio = 6.81; 95% confidence interval, 1.10 to 46.86; p < 0.05 and waist circumference odds ratio = 4.34; 95% confidence interval, 1.47 to 12.83; p = 0.001). DISCUSSION: Women who were overweight or had a higher waist circumference (women > or = 88 cm and men > or = 102 cm) have a significantly higher risk of having hyperleptinemia. The increase in age as an isolated variable is not a risk factor for hyperleptinemia. PMID- 11943832 TI - Alcohol consumption in the severely obese: relationship with the metabolic syndrome. AB - OBJECTIVE: The aim of this study was to examine the association between the clinical and biochemical features of the metabolic syndrome and quantity and type of alcohol intake in the severely obese. RESEARCH METHODS AND PROCEDURES: A cross sectional study was performed in 486 consecutive severely obese subjects. Data on alcohol consumption was collected by serial clinical interviews and a questionnaire. The relationship between alcohol intake and the clinical and serum chemistry features of the metabolic syndrome was analyzed by multiple statistical techniques. Laboratory measures included lipid profile, fasting blood glucose, hemoglobin A1c, and fasting serum insulin. An indirect index of insulin resistance was calculated using the log-transformed fasting insulin and glucose product. RESULTS: There were 486 subjects, 84% women, with a mean age of 40.6 +/- 10 years (range, 16 to 71 years) and a body mass index of 45.3 +/- 7 kg/m(2) (range, 34 to 77 kg/m(2)). Alcohol consumers (N = 276) showed a marked reduction in the adjusted odds ratio of type 2 diabetes (odds ratio = 0.29; 95% confidence interval, 0.16 to 0.55) compared with rare or nonconsumers (N = 210). There was a U-shaped relationship between the amount and frequency of alcohol consumption and fasting triglyceride, fasting glucose, hemoglobin A1c, and index of insulin resistance measurements. Consumers of < 100 g/wk had more favorable measures. The effect was attenuated when diabetics were excluded from the analysis. Timing of alcohol consumption did not influence outcome measures. DISCUSSION: Light-to moderate alcohol consumption is associated with a lower prevalence of type 2 diabetes, reduced insulin resistance, and more favorable vascular risk profile in the severely obese. We would propose that light to moderate alcohol consumption should not be discouraged in the severely obese. PMID- 11943834 TI - Regional intra-subject variability in abdominal adiposity limits usefulness of computed tomography. AB - OBJECTIVE: Computed tomography (CT) and magnetic resonance imaging, the most accurate methods of abdominal fat measurement, have been applied using a number of protocols, ranging from single-slice area determination to multiple-slice volume calculation. The aim of this study was to assess the validity of single slice CT for abdominal fat area measurement by estimating the intra-subject variability in abdominal fat areas and comparing the ranking of subjects across four contiguous abdominal levels. RESEARCH METHODS AND PROCEDURES: Nineteen premenopausal women (age, 35.3 +/- 1.4 years; mean +/- SE) were studied. CT was used to measure intra-abdominal fat (IAF) area, percentage of total intra abdominal area (%IAF), subcutaneous abdominal fat (SAF) area, and IAF/SAF at four adjacent cross-sectional lumbar levels (L2-L4). Intra-subject variability (percentage) was defined as SD/mean x 100. Total body fat was measured by DXA, which was further analyzed for central abdominal fat. RESULTS: Mean body mass index was 24.9 +/- 1.0 kg/m(2). The average (range) intra-subject variability was 28% (8% to 61%) for IAF, 46% (19% to 124%) for %IAF, 26% (14% to 38%) for SAF area, and 19% (7% to 71%) for IAF/SAF. The pattern of this variability was not uniform between subjects, because their ranking by IAF area was markedly different at each CT level. DISCUSSION: We demonstrated significant intra-subject variability in CT-measured adipose tissue areas across four predetermined sites. This resulted in a difference in the ordering of subjects by IAF at each of the four imaging sites, suggesting that the usefulness of single-slice CT in the assessment of abdominal adiposity in premenopausal women may be limited, particularly when performed for the purpose of making comparisons between subjects based on abdominal fat area. PMID- 11943835 TI - Lipolysis in adipocytes isolated from deep and superficial subcutaneous adipose tissue. AB - OBJECTIVE: Abdominal subcutaneous adipose tissue (SAT) occurs in two depots separated by a fascial plane: deep SAT and superficial SAT. In a recent study it was demonstrated that the amount of deep SAT has a much stronger relationship to insulin resistance than does superficial SAT. Because insulin resistance may be related to fatty acid release from adipose tissue, we hypothesized that the two SAT depots may have a different lipolytic activity. RESEARCH METHODS AND PROCEDURES: To test this hypothesis, we obtained samples of deep and superficial SAT from patients undergoing elective abdominal surgery. The rate of lipolysis was determined in the collagenase-digested adipocytes obtained from the two fat depots by measuring glycerol release in the presence and absence of isoproterenol. In addition, the relative concentration of hormone-sensitive lipase was determined in both SAT depots by Western blot analysis. RESULTS: Our results showed that the rate of isoproterenol-stimulated lipolysis was approximately 20% higher in cells from deep SAT compared with those from superficial SAT, indicating that the deep SAT is more lipolytically active. The concentration of hormone-sensitive lipase did not differ between the two adipose tissue depots. DISCUSSION: These findings suggest that the higher lipolytic activity of deep SAT may account for its stronger association with insulin resistance. The mechanism seems to be independent of differences in hormone sensitive lipase concentration. PMID- 11943836 TI - Work-related physical activity is not associated with body mass index and obesity. AB - OBJECTIVE: To analyze the association of work-related physical activity (WRPA) and leisure-time physical activity (LTPA) with body mass index (BMI) and obesity in the Spanish adult population aged 20 to 60 years. RESEARCH METHODS AND PROCEDURES: The data were taken from the 1993 Spanish National Health Survey. We analyzed a sample of 12,044 men and women representative of the Spanish population aged 20 to 60 years. BMI and frequency of obesity (BMI > or = 30 kg/m(2)) were obtained from self-reported weight and height. Multiple linear regression and logistic regression models were constructed, adjusting for the main confounding factors. WRPA and LTPA were measured by two questions to classify subjects into four categories of physical activity. RESULTS: Neither mean BMI nor percentage of obesity varied significantly (p > 0.05) by WRPA. Mean BMI was significantly higher (p < 0.01) in those who were inactive in their leisure time (25.90 kg/m(2) in men and 24.43 kg/m(2) in women) than in those who reported vigorous activity (24.42 kg/m(2) and 22.97 kg/m(2) in men and women, respectively). The odds ration (OR) for obesity decreased with increasing level of LTPA in both men (OR of 0.64 for vigorous activity) and women (OR = 0.68), showing a statistically significant dose-response relation in both men (for linear trend, p = 0.0021) and women (p = 0.0245). DISCUSSION: These results raise questions about the association between WRPA and obesity and suggest the need to reexamine models of the obesity epidemic that point to automation of the workplace as one of the major explanatory factors. PMID- 11943837 TI - The road to obesity or the path to prevention: motorized transportation and obesity in China. AB - OBJECTIVE: Dependence on motorized forms of transportation may contribute to the worldwide obesity epidemic. Shifts in transportation patterns occurring in China provide an ideal opportunity to study the association between vehicle ownership and obesity. Our objective was to determine whether motorized forms of transportation promote obesity. RESEARCH METHODS AND PROCEDURES: A multistage random-cluster sampling process was used to select households from eight provinces in China. Data were included on household vehicle ownership and individual anthropometric and sociodemographic status. Cross-sectional data (1997) from 4741 Chinese adults aged 20 to 55 years were used to explore the association between vehicle ownership and obesity. Cohort data (1989 to 1997) from 2485 adults aged 20 to 45 years in 1989 (59% follow-up) were used to measure the impact of vehicle acquisition on the odds of becoming obese. RESULTS: Our main outcome measure was current obesity status and the odds of becoming obese over an 8-year period. In 1997, 84% of adults did not own motorized transportation. However, the odds of being obese were 80% higher (p < 0.05) for men and women in households who owned a motorized vehicle compared with those who did not own a vehicle. Fourteen percent of households acquired a motorized vehicle between 1989 and 1997. Compared with those whose vehicle ownership did not change, men who acquired a vehicle experienced a 1.8-kg greater weight gain (p < 0.05) and had 2 to 1 odds of becoming obese. DISCUSSION: Encouraging active forms of transportation may be one way to protect against obesity. PMID- 11943839 TI - Modeling the impact of adjustable gastric banding on survival in patients with morbid obesity. AB - OBJECTIVE: Morbid obesity is associated with premature death. Adjustable gastric banding may lead to substantial weight loss in patients with morbid obesity. Little is known about the impact of weight loss on survival after adjustable gastric banding. We therefore developed a mathematical model to estimate life expectancy in patients with a body mass index (BMI) > or =40 kg/m(2) undergoing bariatric surgery. RESEARCH METHODS AND PROCEDURES: We developed a nonhomogeneous Markov chain consisting of five states: the absorbing state ("dead") and the four recurrent states BMI > or =40 kg/m(2), BMI 36 to 39 kg/m(2), BMI 32 to 35 kg/m(2), and BMI 25 to 31 kg/m(2). Scenarios of weight loss and age- and sex dependent risk of death, as well as BMI-dependent excess mortality were extracted from life tables and published literature. All patients entered the model through the state of BMI > or =40 kg/m(2). RESULTS: In men aged either 18 or 65 years at the time of surgery, who moved from the state BMI > or =40 kg/m(2) to the next lower state of BMI 36 to 39 kg/m(2), life expectancy increased by 3 and 0.7 years, respectively. In women aged either 18 or 65 years at the time of surgery, who moved from the state BMI > or =40 kg/m(2) to the next lower state BMI 36 to 39 kg/m(2), life expectancy increased by 4.5 and 2.6 years, respectively. Weight loss to lower BMI strata resulted in further gains of life expectancy in both men and women. DISCUSSION: Within the limitations of the modeling study, adjustable gastric banding in patients with morbid obesity may substantially increase life expectancy. PMID- 11943838 TI - Age dimorphism in the association between growth-hormone status and the respiratory quotient. AB - OBJECTIVE: To investigate the impact of age on the association between the respiratory quotient (RQ) and growth-hormone (GH) secretion and to investigate the acute lipolytic response to an exogenous GH bolus. RESEARCH METHODS AND PROCEDURES: A cross-sectional study of 36 non-obese healthy subjects (18 women and 18 men) from two age groups was used: "younger" (mean age, 29.5 years; range, 27 to 34 years) and "older" (mean age, 50.8 years; range, 47 to 59 years). Endogenous GH secretion by means of deconvolution analysis of 24-hour serum GH concentrations was measured every 20 minutes. Resting RQ was measured after a 12 hour overnight fast. The lipolytic response to an intravenous exogenous GH bolus (200 microg) was assessed by measuring serum levels of free fatty acids as well as changes in RQ. Additional measurements included body composition (regional computed tomography scan and DXA) and physical fitness (VO(2)max). RESULTS: Resting RQ did not differ between the two age groups: 0.81 +/- 0.01 (young) vs. 0.82 +/- 0.01 (older; not significant). Several estimates of GH release correlated positively with RQ in the younger group, whereas a negative correlation was detected in the older subjects [GH production rate (microg/liter x kg) vs. RQ: r = 0.62, p < 0.01 (younger); r = -0.53; p = 0.02 (older)]. By regression analysis, 52% to 58% of the variation in RQ could be explained by GH status. After an exogenous GH bolus, the incremental response in nonesterified fatty acid was slightly higher in younger individuals (p = 0.09). DISCUSSION: Resting RQ is significantly correlated with GH status. This association is positive in younger individuals and negative in older individuals. The lipolytic response to exogenous GH is moderately higher in younger compared with older individuals. GH status should be taken into account when investigating the residual variation in RQ. PMID- 11943840 TI - Gender effects on adrenergic receptor expression and lipolysis in white adipose tissue of rats. AB - OBJECTIVE: To investigate the effects of short-term (15 days) cafeteria-diet feeding on the expression of alpha- and beta-adrenergic receptors (AR) and its association with lipolytic stimulation in isolated retroperitoneal white adipocytes. RESEARCH METHODS AND PROCEDURES: Six female and 6 male Wistar rats (4 weeks old) were fed a cafeteria diet plus standard diet for 15 days. The remaining 12 age- and sex-matched rats received a standard diet only. White retroperitoneal adipose tissue was isolated and used for the determination of both alpha(2) and beta-AR expression and for in vitro studies of lipolytic activity. RESULTS: In female control rats, we found higher lipolytic capacities located at the postreceptor level and a lower alpha(2)/beta(3)-AR ratio than male rats. Cafeteria-diet feeding for 15 days decreased lipolytic activity in both male and female rats and altered the alpha(2A)- and beta(3)-AR protein levels with an increase of alpha(2A)-AR in males and a beta(3)-AR decrease in females. DISCUSSION: Our results indicate that a 15-day cafeteria-diet feeding induced an increase in the alpha(2)/beta(3)-AR balance and impaired adipose tissue lipolytic activity, which was higher in males and may contribute to the development of increased fat mass. The higher functionality of alpha(2)-AR, together with the minor role developed by beta(3)-AR and lower lipolytic capacities located at the postreceptor level in cafeteria-diet-fed male rats compared with female rats, may be responsible for the gender-dependent differences observed in this study. PMID- 11943841 TI - Leptin enhances adenosine diphosphate-induced platelet aggregation in healthy subjects. PMID- 11943842 TI - Polymorphonuclear leukocyte membrane fluidity and insulin resistance in obese subjects. PMID- 11943843 TI - Somatosensory feedback modulates the respiratory motor program of crystallized birdsong. AB - Birdsong, like human speech, involves rapid, repetitive, or episodic motor patterns requiring precise coordination between respiratory, vocal organ, and vocal tract muscles. The song units or syllables of most adult songbirds exhibit a high degree of acoustic stereotypy that persists for days or months after the elimination of auditory feedback by deafening. Adult song is assumed to depend on central motor programs operating independently from immediate sensory feedback. Nothing is known, however, about the possible role of mechanoreceptive or other somatosensory feedback in the motor control of birdsong. Even in the case of human speech, the question of "how and when sensory information is used in normal speaking conditions...remains unanswered" and controversial [Smith, A. (1992) Crit. Rev. Oral Biol. Med. 3, 233-267]. We report here evidence for somatosensory modulation of ongoing song motor patterns. These patterns include the respiratory muscles that, in both birdsong and speech, provide the power for vocalization. Perturbing respiratory pressure by a brief, irregularly timed injection of air into the cranial thoracic air sac during song elicited a compensatory reduction in the electrical activity of the abdominal expiratory muscles, both in hearing and deafened adult northern cardinals (Cardinalis cardinalis). This muscle response was absent or reduced during quiet respiration, suggesting it is specifically linked to phonation. Our findings indicate that somatosensory feedback to expiratory muscles elicits compensatory adjustments that help stabilize, in real time, the subsyringeal pressure against fluctuations caused by changes in posture or physical activity. PMID- 11943844 TI - Dechlorination of chloroacetanilide herbicides by thiosulfate salts. AB - Halogenated organic compounds (XOCs) are among the most widely used synthetic chemicals. Many XOCs are recalcitrant to natural degradation and have become prominent environmental contaminants. One group of such XOCs are the heavily used chloroacetanilide herbicides. We have found that chloroacetanilide herbicides are rapidly dechlorinated in water, sand, and soil by thiosulfate salts under ambient conditions. Structural and kinetics analysis suggests that the reaction occurred by S(N)2 nucleophilic substitution, in which the chlorine was replaced by thiosulfate and the herbicide was detoxified. Laboratory studies showed that this reaction could be used for removing residues of chloroacetanilide herbicides in water, soil, and sand. Our findings also suggest that some other XOCs may be subject to this reaction. Because common thiosulfate salts are innocuous products (e.g., fertilizers) and the reaction selectively detoxifies XOCs at low thiosulfate levels, this discovery may lead to a new way for safe removal of certain XOCs from the environment. PMID- 11943846 TI - Impaired neural tube closure, axial skeleton malformations, and tracheal ring disruption in TRAF4-deficient mice. AB - TRAF4 belongs to the tumor necrosis factor receptor-associated factor (TRAF) family of proteins but, unlike other family members, has not yet been clearly associated to any specific receptor or signaling pathway. To investigate the biological function of TRAF4, we have generated traf4-deficient mice by gene disruption. The traf4 gene mutation is embryonic lethal but with great individual variation, as approximately one third of the homozygous mutant embryos died in utero around embryonic day 14, whereas the others reach adulthood. Surviving mutant mice manifest numerous developmental abnormalities; notably, 100% of homozygous mutant mice suffer respiratory disorder and wheezing caused by tracheal ring disruption. Additional malformations concern mainly the axial skeleton, as the ribs, sternum, tail, and vertebral arches are affected, with various degrees of penetrance. Traf4-deficient mice also exhibit a high incidence of spina bifida, a defect likened to neural tube defects (NTD) that are common congenital malformations in humans. Altogether, our results demonstrate that TRAF4 is required during embryogenesis in key biological processes including the formation of the trachea, the development of the axial skeleton, and the closure of the neural tube. Considering the normal expression pattern of TRAF4 in neural tissues, we can conclude that TRAF4 participates in neurulation in vivo. PMID- 11943847 TI - Molecular evolution of the HoxA cluster in the three major gnathostome lineages. AB - The duplication of Hox clusters and their maintenance in a lineage has a prominent but little understood role in chordate evolution. Here we examined how Hox cluster duplication may influence changes in cluster architecture and patterns of noncoding sequence evolution. We sequenced the entire duplicated HoxAa and HoxAb clusters of zebrafish (Danio rerio) and extended the 5' (posterior) part of the HoxM (HoxA-like) cluster of horn shark (Heterodontus francisci) containing the hoxa11 and hoxa13 orthologs as well as intergenic and flanking noncoding sequences. The duplicated HoxA clusters in zebrafish each house considerably fewer genes and are dramatically shorter than the single HoxA clusters of human and horn shark. We compared the intergenic sequences of the HoxA clusters of human, horn shark, zebrafish (Aa, Ab), and striped bass and found extensive conservation of noncoding sequence motifs, i.e., phylogenetic footprints, between the human and horn shark, representing two of the three gnathostome lineages. These are putative cis-regulatory elements that may play a role in the regulation of the ancestral HoxA cluster. In contrast, homologous regions of the duplicated HoxAa and HoxAb clusters of zebrafish and the HoxA cluster of striped bass revealed a striking loss of conservation of these putative cis-regulatory sequences in the 3' (anterior) segment of the cluster, where zebrafish only retains single representatives of group 1, 3, 4, and 5 (HoxAa) and group 2 (HoxAb) genes and in the 5' part of the clusters, where zebrafish retains two copies of the group 13, 11, and 9 genes, i.e., AbdB-like genes. In analyzing patterns of cis-sequence evolution in the 5' part of the clusters, we explicitly looked for evidence of complementary loss of conserved noncoding sequences, as predicted by the duplication-degeneration-complementation model in which genetic redundancy after gene duplication is resolved because of the fixation of complementary degenerative mutations. Our data did not yield evidence supporting this prediction. We conclude that changes in the pattern of cis-sequence conservation after Hox cluster duplication are more consistent with being the outcome of adaptive modification rather than passive mechanisms that erode redundancy created by the duplication event. These results support the view that genome duplications may provide a mechanism whereby master control genes undergo radical modifications conducive to major alterations in body plan. Such genomic revolutions may contribute significantly to the evolutionary process. PMID- 11943848 TI - NMDA receptor function is regulated by the inhibitory scaffolding protein, RACK1. AB - Phosphorylation regulates the function of ligand-gated ion channels such as the N methyl d-aspartate (NMDA) receptor. Here we report a mechanism for modulation of the phosphorylation state and function of the NMDA receptor via an inhibitory scaffolding protein, RACK1. We found that RACK1 binds both the NR2B subunit of the NMDA receptor and the nonreceptor protein tyrosine kinase, Fyn. RACK1 inhibits Fyn phosphorylation of NR2B and decreases NMDA receptor-mediated currents in CA1 hippocampal slices. Peptides that disrupt the interactions between RACK1, NR2B, and Fyn induce phosphorylation and potentiate NMDA receptor mediated currents. Therefore, RACK1 is a regulator of NMDA receptor function and may play a role in synaptic plasticity, addiction, learning, and memory. PMID- 11943849 TI - Conformation and anion binding properties of cyclic hexapeptides containing L-4 hydroxyproline and 6-aminopicolinic acid subunits. AB - Two cyclic hexapeptides containing alternating all R and all S configured Ll (4R/S)-hydroxyproline and 6-aminopicolinic acid subunits are presented, and the influence of the hydroxyl groups on the solubility, conformation, and receptor properties is investigated. Cyclopeptide 2, containing the natural 4R configured hydroxyproline, adopts a conformation similar to that of the unsubstituted peptide 1, which is able to bind anions such as halides and sulfate in aqueous solution. 2 also interacts with these anions, but whereas 1 forms sandwich type 2:1 complexes, in which the anion is bound by two cyclopeptide moieties, 2 forms 1:1 complexes. The stabilities of the halide and sulfate complexes of 2 range between 10(0) and 10(2) M(-1) in 80% D(2)O/CD(3)OD. Complex formation is detectable even in water, but with slightly smaller stability constants. Using this information a quantitative evaluation of the stability of the 2:1 complexes of 1, for which overall stability constants in the order 10(4) to 10(5) M(-2) in 80% D(2)O/CD(3)OD were observed, was made. In contrast to 2, the conformation of 3, containing the non-natural 4S configured hydroxyproline, is strongly affected by the presence of the hydroxyl groups. In d(6)-DMSO and methanol/water mixtures a slow conformational equilibrium between two C(3)-symmetrical conformers is observed, and 3 is thus much less preorganized for anion binding than either 1 or 2. PMID- 11943850 TI - Self-organization of self-assembled photonic materials into functional devices: photo-switched conductors. AB - Linear porphyrin arrays self-assembled by either hydrogen bonding or metal ion coordination self-organize into lipid bilayer membranes. The length of the transmembrane assemblies is determined both by the thermodynamics of the intermolecular interactions in the supermolecule and by the dimension and physical chemical properties of the bilayer. Thus, the size of the porphyrin assembly can self-adjust to the thickness of the bilayer. An aqueous electron acceptor is placed on one side of the membrane and an electron donor is placed on the opposite side. When illuminated with white light, substantial photocurrents are observed. Only the assembled structures give rise to the photocurrent, as no current is observed from any of the component molecules. The fabrication of this photogated molecular electronic conductor from simple molecular components exploits several levels of self-assembly and self-organization. PMID- 11943851 TI - Partial shell-filled core-shell tecto(dendrimers): a strategy to surface differentiated nano-clefts and cusps. AB - Poly(amidoamine) (PAMAM) dendrimer shell reagents possessing either nucleophilic (i.e., primary amines) or electrophilic (i.e., carboxymethyl esters) functional groups have been covalently assembled around appropriate electrophilic or nucleophilic dendrimer core reagents to produce partial shell filled/core-shell tecto(dendrimers). Partial shell-filled products with saturation levels ranging from 28% to 66% were obtained. These metastable, remarkably monodispersed assemblies possess functionally differentiated nano-cusps and clefts that exhibit "autoreactive" behavior. Pacification of these autoreactive products with appropriate alkanolamine reagents produced robust, nonreactive, "hydroxy-amine differentiated" surfaces that exhibit very active self-assembly properties. Based on the monodispersity, dimensional scaling, and electrophoretic similarities of PAMAM dendrimers to globular proteins, these assemblies may be viewed as crude biomimetics of classical core shell-type protein aggregates. These dimensionally larger, but analogous PAMAM core-shell tecto(dendrimer) architectures extend and complete a similar pattern of autoreactivity and pacification that was observed earlier for traditional mono PAMAM dendrimer core-shell modules possessing unsaturated shell levels. PMID- 11943852 TI - Analysis of structure and function relationships of an autoantigenic peptide of insulin bound to H-2K(d) that stimulates CD8 T cells in insulin-dependent diabetes mellitus. AB - The recognition of MHC-peptide complexes by T cells is governed by structural considerations that are determined by the sequences of the individual components and their interaction with each other. We have studied the function of a highly diabetogenic CD8 T cell clone that is specific for insulin B15-23:H-2K(d). We have then related this to modeled MHC-peptide structures. The native peptide binds poorly to H-2K(d), because of the small glycine residue at peptide position p9 that is incapable of productive interactions with the hydrophobic residues of pocket F. In addition, electrostatic repulsions between the peptide glutamate residue at position 7 and 152D of the MHC molecule heavy chain contribute to the poor binding. However, B chain peptide 15-23 bound to K(d) shows excellent T cell stimulation and the induction of CD8 cytotoxic T cells. Peptide substitution has also shown that p6G is likely to be a T cell antigen receptor interaction site. Our studies have shown that the predictions seen in the models correlate closely with the observed effects in functional assays and provide insight into how this peptide, which would not be predicted to stimulate these cells on H-2K(d) binding studies alone, could activate such highly pathogenic T cells. PMID- 11943853 TI - Somatosensory cortex dominated by the representation of teeth in the naked mole rat brain. AB - We investigated naked mole-rat somatosensory cortex to determine how brain areas are modified in mammals with unusual and extreme sensory specializations. Naked mole-rats (Heterocephalus glaber) have numerous anatomical specializations for a subterranean existence, including rows of sensory hairs along the body and tail, reduced eyes, and ears sensitive to low frequencies. However, chief among their adaptations are behaviorally important, enlarged incisors permanently exterior to the oral cavity that are used for digging, object manipulation, social interactions, and feeding. Here we report an extraordinary brain organization where nearly one-third (31%) of primary somatosensory cortex is devoted to the representations of the upper and lower incisors. In addition, somatosensory cortex is greatly enlarged (as a proportion of total neocortical area) compared with closely related laboratory rats. Finally, somatosensory cortex in naked mole rats encompasses virtually all of the neocortex normally devoted to vision. These findings indicate that major cortical remodeling has occurred in naked mole-rats, paralleling the anatomical and behavioral specializations related to fossorial life. PMID- 11943854 TI - Activation of mislocalized opsin kills rod cells: a novel mechanism for rod cell death in retinal disease. AB - Rod photoreceptors are highly compartmentalized sensory neurons that maintain strict ultrastructural and molecular polarity. Structural subdivisions include the outer segment, inner segment, cell body, and synaptic terminal. The visual pigment rhodopsin is found predominantly in membranes of the rod cell outer segment but becomes mislocalized, appearing throughout the plasma membrane of the cell in many retinal diseases and injuries. Currently, there is no known link between rhodopsin redistribution and rod cell death. We propose that activation of mislocalized rhodopsin kills rod cells by stimulating normally inaccessible signaling pathways. This hypothesis was tested in primary retinal cell cultures, which contain photoreceptors. In rod photoreceptors, opsin immunofluorescence occurred throughout the rod cell plasma membrane. Activation of this mislocalized opsin by photostimulation after formation of isorhodopsin or by incubation with beta-ionone (opsin agonist) killed 19-30% of rod cells. Rod cell death was apoptotic, as indicated by marked chromatin condensation and the requirement for caspase-3 activation. Rod cell death could be induced by forskolin (adenylate cyclase agonist), and conversely, beta-ionone-induced cell death could be blocked by cotreatment with SQ22536 (an adenylate cyclase inhibitor). Pertussis toxin (a G protein inhibitor) also blocked beta-ionone-induced cell death. The data support a mechanism by which activation of mislocalized opsin initiates apoptotic rod cell death through G protein stimulation of adenylate cyclase. PMID- 11943855 TI - Extending supramolecular fullerene-porphyrin chemistry to pillared metal-organic frameworks. AB - Porphyrins and fullerenes are spontaneously attracted to each other. This supramolecular recognition element can be exploited to produce ordered arrays of interleaved porphyrins and fullerenes. C(60) x H(2)TpyP x Pb(NO(3))(2) x 1.5TCE (H(2)TpyP = tetra-4-pyridylporphyrin; TCE = 1,1,2,2-tetrachloroethane) crystallizes in the tetragonal P4/n space group and the structure has been solved to high resolution. The Pb(2+) ions connect the pyridylporphyrins in infinite sheets with an interlayer spacing of 12.1 A. The fullerenes are intercalated between these layers, acting as pillars. The 6:6 ring juncture bonds of C(60) are centered over the porphyrins, bringing the layers into strict tetragonal register. This arrangement identifies the fullerene-porphyrin interaction as a structure-defining element. The same motif is seen in a related ribbon structure having C(70) intercalated into HgI(2)-linked H(2)TpyTP. The supramolecular design principles involved in assembling these chromophores may have applications in materials science. PMID- 11943856 TI - Design and self-assembly of wide and robust coordination cages. AB - The self-assembly of a new class of coordination cages formed from two tetrapyridyl-substituted cavitands connected through four square-planar palladium or platinum complexes is reported. The shape of the internal cavity resembles an ellipsoid with a calculated volume of 840 A(3). The four lateral portals have a diameter of about 6 A, large enough to allow the fast entrance/egress of counterions in solution. The platinum cages 3a,e cannot be disassembled using triethylamine as competitive ligand and they are kinetically stable at room temperature, whereas the palladium cages 3b-d, 3f-h are disassembled and kinetically labile under the same conditions. The different solubility properties of the cage components have allowed the extension of this self-assembly protocol to the liquid-liquid interface. PMID- 11943857 TI - A peptide template as an allosteric supramolecular catalyst for the cleavage of phosphate esters. AB - The heptapeptide H-Iva-Api-Iva-ATANP-Iva-Api-Iva-NHCH(3) (P1a), where Iva is (S) isovaline, Api is 4-amino-4-carboxypiperidine, and ATANP is (S)-2-amino-3-[1 (1,4,7-triazacyclononane)]propanoic acid, has been synthesized. Its conformation in aqueous solution is essentially that of a 3(10)-helix. By connecting three copies of P1a to a functionalized Tris(2-aminoethyl)amine (Tren) platform a new peptide template, [T(P1)(3)], was obtained. This molecule is able to bind up to four metal ions (Cu(II) or Zn(II)): one in the Tren subsite and three in the azacyclononane subunits. The binding of the metals to the Tren platform induces a change from an open to a closed conformation in which the three short, helical peptides are aligned in a parallel manner with the azacyclonane units pointing inward within the pseudocavity they define. T(P1)(3) shows a peculiar behavior in the transphosphorylation of phosphate esters; the tetrazinc complex is a catalyst of the cleavage of 2-hydroxypropyl-p-nitrophenyl phosphate (HPNP), whereas the free ligand is a catalyst of the cleavage of an oligomeric RNA sequence with selectivity for pyrimidine bases. In the case of HPNP, Zn(II) acts as a positive allosteric effector by enhancing the catalytic efficiency of the system. In the case of the polyanionic RNA substrate, Zn(II) switches off the activity, thus behaving as a negative allosteric regulator. It is suggested that the opposite behavior of the catalyst induced by Zn(II) is associated with the change of conformation of the Tren platform, and consequently of the relative spatial disposition of the three linked peptides, that occurs after binding of the metal ion. PMID- 11943858 TI - Chiral discrimination in cyclodextrin complexes of amino acid derivatives: beta cyclodextrin/N-acetyl-L-phenylalanine and N-acetyl-D-phenylalanine complexes. AB - In a systematic study of molecular recognition of amino acid derivatives in solid state beta-cyclodextrin (beta-CD) complexes, we have determined crystal structures for complexes of beta-cyclodextrin/N-acetyl-L-phenylalanine at 298 and 20 K and for N-acetyl-D-phenylalanine at 298 K. The crystal structures for the N acetyl-L-phenylalanine complex present disordered inclusion complexes for which the distribution of guest molecules at room temperature is not resolvable; however, they can be located with considerable confidence at low temperature. In contrast, the complex with N-acetyl-D-phenylalanine is well ordered at room temperature. The latter complex presents an example of a complex in this series in which a water molecule is included deeply in the hydrophobic torus of the extended dimer host. In an effort to understand the mechanisms of molecular recognition giving rise to the dramatic differences in crystallographic order in these crystal structures, we have examined the intermolecular interactions in detail and have examined insertion of the enantiomer of the D-complex into the chiral beta-CD complex crystal lattice. PMID- 11943859 TI - A structure-based method for derivation of all-atom potentials for protein folding. AB - A method for deriving all-atom protein folding potentials is presented and tested on a three-helix bundle protein, as well as on hairpin and helical sequences. The potentials obtained are composed of a contact term between pairs of atoms, and a local density term for each atom, mimicking solvent exposure preferences. Using this potential in an all-atom protein folding simulation, we repeatedly folded the three-helix bundle, with the lowest energy conformations having a C(alpha) distance rms from the native structure of less than 2 A. Similar results were obtained for the hairpin and helices by using different potentials. We derived potentials for several different proteins and found a high correlation between the derived parameters, suggesting that a potential of this form eventually could be found that folds multiple, unrelated proteins at the atomic level of detail. PMID- 11943861 TI - Threefold symmetric trimetallic macrocycles formed via the Weak-Link Approach. AB - The synthesis and characterization of a new threefold symmetric hemilabile phosphino-alkylthioether ligand are described. This ligand can be used in combination with Rh(I) and Ir(I) precursor complexes to prepare 40-membered macrocyles with threefold symmetry via the Weak-Link Approach. Synthesis and characterization of two such structures are reported along with a single crystal x-ray diffraction analysis of one of the key intermediates in the case of Rh(I). This is a demonstration of the viability of the Weak-Link Approach for preparing structures other than bimetallic macrocycles and suggests that it could be generalized for a wide range of higher symmetry structures through appropriate hemilabile ligand design. PMID- 11943860 TI - Clonally expanded mtDNA point mutations are abundant in individual cells of human tissues. AB - Using single-cell sequence analysis, we discovered that a high proportion of cells in tissues as diverse as buccal epithelium and heart muscle contain high proportions of clonal mutant mtDNA expanded from single initial mutant mtDNA molecules. We demonstrate that intracellular clonal expansion of somatic point mutations is a common event in normal human tissues. This finding implies efficient homogenization of mitochondrial genomes within individual cells. Significant qualitative differences observed between the spectra of clonally expanded mutations in proliferating epithelial cells and postmitotic cardiomyocytes suggest, however, that either the processes generating these mutations or mechanisms driving them to homoplasmy are likely to be fundamentally different between the two tissues. Furthermore, the ability of somatic mtDNA mutations to expand (required for their phenotypic expression), as well as their apparently high incidence, reinforces the possibility that these mutations may be involved actively in various physiological processes such as aging and degenerative disease. The abundance of clonally expanded point mutations in individual cells of normal tissues also suggests that the recently discovered accumulation of mtDNA mutations in tumors may be explained by processes that are similar or identical to those operating in the normal tissue. PMID- 11943862 TI - Experimental analysis of sperm competition mechanisms in a wild bird population. AB - Recent work on wild birds has revealed the importance of sperm competition as a source of sexual selection, but behavioral and paternity studies have previously provided only indirect evidence for mechanisms of sperm competition in wild birds. In a field study of collared flycatchers Ficedula albicollis we used a previously uncharacterized method to determine the frequency and timing of extra pair inseminations. By counting the number of sperm trapped on the perivitelline layer of eggs, we determined the timing of inseminations and estimated, on a day to-day basis, the amount of sperm females stored. Our results showed that female collared flycatchers preferentially engaged in extra-pair copulations when mated to an unattractive male with a small white forehead patch. These copulations were timed for the middle part of their fertile period, at least 2 days after the last within-pair insemination. Although the mean number of extra-pair insemination events was only 1.33 per cuckolding female, the ratio between the number of sperm from extra-pair and pair inseminations was at least 5 to 1. Thus a single, well timed extra-pair insemination caused by female behavior could greatly bias fertilization probability in favor of an attractive extra-pair male. Our results suggest a possible behavioral mechanism for female control of sperm competition. PMID- 11943863 TI - Mechanism of plasmid-mediated quinolone resistance. AB - Quinolones are potent antibacterial agents that specifically target bacterial DNA gyrase and topoisomerase IV. Widespread use of these agents has contributed to the rise of bacterial quinolone resistance. Previous studies have shown that quinolone resistance arises by mutations in chromosomal genes. Recently, a multiresistance plasmid was discovered that encodes transferable resistance to quinolones. We have cloned the plasmid-quinolone resistance gene, termed qnr, and found it in an integron-like environment upstream from qacE Delta 1 and sulI. The gene product Qnr was a 218-aa protein belonging to the pentapeptide repeat family and shared sequence homology with the immunity protein McbG, which is thought to protect DNA gyrase from the action of microcin B17. Qnr had pentapeptide repeat domains of 11 and 28 tandem copies, separated by a single glycine with a consensus sequence of A/C D/N L/F X X. Because the primary target of quinolones is DNA gyrase in Gram-negative strains, we tested the ability of Qnr to reverse the inhibition of gyrase activity by quinolones. Purified Qnr-His(6) protected Escherichia coli DNA gyrase from inhibition by ciprofloxacin. Gyrase protection was proportional to the concentration of Qnr-His(6) and inversely proportional to the concentration of ciprofloxacin. The protective activity of Qnr-His(6) was lost by boiling the protein and involved neither quinolone inactivation nor independent gyrase activity. Protection of topoisomerase IV, a secondary target of quinolone action in E. coli, was not evident. How Qnr protects DNA gyrase and the prevalence of this resistance mechanism in clinical isolates remains to be determined. PMID- 11943864 TI - Beneficial effects of systemic administration of recombinant human erythropoietin in rabbits subjected to subarachnoid hemorrhage. AB - Cerebral vasospasm and ischemic damage are important causes of mortality and morbidity in patients affected by aneurysmal subarachnoid hemorrhage (SAH). Recently, i.p. administration of recombinant human erythropoietin (r-Hu-EPO) has been shown to exert a neuroprotective effect during experimental SAH. The present study was conducted to evaluate further the effect of r-Hu-EPO administration after SAH in rabbits on neurological outcome, degree of basilar artery spasm, and magnitude of neuronal ischemic damage. Experimental animals were divided into six groups: group 1 (n = 8), control; group 2 (n = 8), control plus placebo; group 3 (n = 8), control plus r-Hu-EPO; group 4 (n = 8), SAH; group 5 (n = 8), SAH plus placebo; group 6 (n = 8), SAH plus r-Hu-EPO. r-Hu-EPO, at a dose of 1,000 units/kg, and placebo were injected i.p. starting 5 min after inducing SAH and followed by clinical and pathological assessment 72 h later. Systemic administration of r-Hu-EPO produced significant increases in cerebrospinal fluid EPO concentrations (P < 0.001), and reduced vasoconstriction of the basilar artery (P < 0.05), ischemic neuronal damage (P < 0.001), and subsequent neurological deterioration (P < 0.05). These observations suggest that r-Hu-EPO may provide an effective treatment to reduce the post-SAH morbidity. PMID- 11943865 TI - Dynamic chemical devices: modulation of contraction/extension molecular motion by coupled-ion binding/pH change-induced structural switching. AB - Dynamic chemical devices involve morphological or constitutional modifications in molecular or supramolecular systems, induced by internal or external physical or chemical triggers. Reversible changes in shape result in molecular motions and define motional dynamic devices presenting mechanical-like actions of various types. Suitably designed polyheterocyclic strands such as compounds 1-5 wrap into helical conformations. The binding of lead(II) ions to the coordination subunits contained in the strand leads to complete uncoiling and yields a polymetallic complex presenting a fully extended shape. The addition of a cryptand complexing agent that strongly binds lead(II) ions and releases them under protonation allows a reversible pH-modulation of lead(II) levels in the medium, which in turn induces coiling/uncoiling of the molecular strand. This system thus represents a motional dynamic device which performs a mechano-chemical process, realizing alternating extension/contraction motions triggered by ion binding. It achieves a linear motor-type of action of very large stroke amplitude fueled by ionic processes. PMID- 11943866 TI - The caspase-cleaved DAP5 protein supports internal ribosome entry site-mediated translation of death proteins. AB - Apoptosis is characterized by a translation switch from cap-dependent to internal ribosome entry site (IRES)-mediated protein translation. During apoptosis, several members of the eukaryotic initiation factor (eIF)4G family are cleaved specifically by caspases. Here we investigated which of the caspase-cleaved eIF4G family members could support cap-independent translation through IRES elements that retain activity in the dying cell. We focused on two major fragments arising from the cleavage of eIF4GI and death-associated protein 5 (DAP5) proteins (eIF4GI M-FAG/p76 and DAP5/p86, respectively), because they are the only potential candidates to preserve the minimal scaffold function needed to mediate translation. Transfection-based experiments in cell cultures indicated that expression of DAP5/p86 in cells stimulated protein translation from the IRESs of c-Myc, Apaf-1, DAP5, and XIAP. In contrast, these IRESs were refractory to the ectopically expressed eIF4GI M-FAG/p76. Furthermore, our study provides in vivo evidence that the caspase-mediated removal of the C-terminal tail of DAP5/p97 relieves an inhibitory effect on the protein's ability to support cap-independent translation through the DAP5 IRES. Altogether, the data suggest that DAP5 is a caspase-activated translation factor that mediates translation through a repertoire of IRES elements, supporting the translation of apoptosis-related proteins. PMID- 11943867 TI - Iron uptake from plasma transferrin by the duodenum is impaired in the Hfe knockout mouse. AB - Hereditary hemochromatosis (HH) is a disorder of iron metabolism in which enhanced iron absorption of dietary iron causes increased iron accumulation in the liver, heart, and pancreas. Most individuals with HH are homozygous for a C282Y mutation in the HFE gene. The function of HFE protein is unknown, but it is hypothesized that it acts in association with beta(2)-microglobulin and transferrin receptor 1 to regulate iron uptake from plasma transferrin by the duodenum, the proposed mechanism by which body iron levels are sensed. The aim of this study was to test this hypothesis by comparing clearance of transferrin bound iron in Hfe knockout (KO) mice with that observed in C57BL/6 control mice. The mice were fed either an iron-deficient, control, or iron-loaded diet for 6 weeks to alter body iron status. The mice then were injected i.v. with (59)Fe transferrin, and blood samples were taken over 2 h to determine the plasma (59)Fe turnover. After 2 h, the mice were killed and the amount of radioactivity in the duodenum, liver, and kidney was measured. In both Hfe KO and C57BL/6 mice, plasma iron turnover and iron uptake from plasma transferrin by the duodenum, liver, and kidney correlated positively with plasma iron concentration. However, duodenal iron uptake from plasma transferrin was decreased in the Hfe KO mice compared with the control mice. Despite this difference in duodenal uptake, the Hfe KO mice showed no decrease in iron uptake by the liver and kidney or alteration in the plasma iron turnover when compared with C57BL/6 mice. These data support the hypothesis that HFE regulates duodenal uptake of transferrin-bound iron from plasma, and that this mechanism of sensing body iron status, as reflected in plasma iron levels, is impaired in HH. PMID- 11943868 TI - Induction and acceleration of insulitis/diabetes in mice with a viral mimic (polyinosinic-polycytidylic acid) and an insulin self-peptide. AB - Polyinosinic-polycytidylic acid (PolyIC), a "mimic" of double-stranded viral RNA, can induce diabetes when administered to rats with RT1(u), and immunization of normal H-2(d) mice (e.g., BALB/c) with insulin B:9-23 peptide (but not H-2(b)) results in the rapid induction of insulin autoantibodies. Because a mouse model of PolyIC/antigen-induced diabetes is lacking, we sought to produce insulitis and diabetes with either PolyIC and/or B:9-23 peptide immunization. Simultaneous administration of PolyIC and B:9-23 peptide to BALB/c mice (but with neither alone) induced insulitis. CD4 T lymphocytes predominated within islets, and the mice did not progress to hyperglycemia. Islets with transgene-induced expression of the costimulatory B7-1 molecule have enhanced diabetes susceptibility. Diabetes was frequently induced in B7-1 transgenic mice with H-2(d) in contrast to H-2(b) mice after PolyIC administration. Disease induction was accelerated by adding B:9-23 immunization to PolyIC. These studies demonstrate that "normal" mice have autoreactive T lymphocytes able to rapidly target islets and insulin given appropriate MHC alleles and that a peripherally administered insulin peptide (an altered peptide ligand of which is in clinical trials) can enhance specific anti-islet autoimmunity. These first PolyIC/insulin-induced murine models should provide an important tool to study the pathogenesis of type 1 diabetes with experimental autoimmune diabetes. PMID- 11943869 TI - Negative regulation of cell growth and differentiation by TSG101 through association with p21(Cip1/WAF1). AB - TSG101 was discovered in a screen for tumor susceptibility genes and has since been shown to have a multiplicity of biological effects. However, the basis for TSG101's ability to regulate cell growth has not been elucidated. We report here that the TSG101 protein binds to the cyclin/cyclin-dependent kinase (CDK) inhibitor (CKI) p21(Cip1/WAF1) and increases stability of the p21 protein in HEK293F cells and differentiating primary keratinocytes, suppressing differentiation in a p21-dependent manner. In proliferating keratinocytes where the p21 protein is relatively stable, TSG101 does not affect the stability or expression of p21 but shows p21-dependent recruitment to cyclin/CDK complexes, inhibits cyclin/CDK activity, and causes strong growth suppression to a much greater extent in p21+/+ than in p21-/- cells. Conversely, suppression of endogenous TSG101 expression by an antisense TSG101 cDNA causes doubling of the fraction of keratinocytes in the S phase of the cell cycle as occurs during p21 deficiency. Our results indicate that TSG101 has a direct role in the control of growth and differentiation in primary epithelial cells, and that p21 is an important mediator of these TSG101 functions. PMID- 11943870 TI - How to assess the relative importance of different colonization routes of pathogens within hospital settings. AB - The emergence of antibiotic resistance among nosocomial pathogens has reemphasized the need for effective infection control strategies. The spread of resistant pathogens within hospital settings proceeds along various routes of transmission and is characterized by large fluctuations in prevalence, which are typical for small populations. Identification of the most important route of colonization (exogenous by cross-transmission or endogenous caused by the selective pressure of antibiotics) is important for the design of optimal infection control strategies. Such identification can be based on a combination of epidemiological surveillance and costly and laborious as well as time consuming methods of genotyping. Furthermore, analysis of the effects of interventions is hampered by the natural fluctuations in prevalence. To overcome these problems, we introduce a mathematical algorithm based on a Markov chain description. The input is longitudinal prevalence data only. The output is estimates of the key parameters characterizing the two colonization routes. The algorithm is tested on two longitudinal surveillance data sets of intensive care patients. The quality of the estimates is determined by comparing them to accurate estimates based on additional information obtained by genotyping. The results warrant optimism that this algorithm may help to quantify transmission dynamics and can be used to evaluate the effects of infection control interventions more carefully. PMID- 11943871 TI - A Kaposi's sarcoma-associated herpesviral protein inhibits virus-mediated induction of type I interferon by blocking IRF-7 phosphorylation and nuclear accumulation. AB - Interferons constitute the earliest immune response against viral infection. They elicit antiviral effects as well as multiple biological responses involved in cell growth regulation and immune activation. Because the interferon-induced cellular antiviral response is the primary defense mechanism against viral infection, many viruses have evolved strategies to antagonize the inhibitory effects of interferon. Here, we demonstrate a strategy that Kaposi's sarcoma associated herpesvirus uses to block virus-mediated induction of type I interferon. We found that a viral immediate-early protein, namely ORF45, interacts with cellular interferon-regulatory factor 7 (IRF-7). In consequence, IRF-7 phosphorylation is inhibited and the accumulation of IRF-7 in the nucleus in response to viral infection is blocked. IRF-7 is a transcription regulator that is responsible for virus-mediated activation of type I interferon genes. By blocking the phosphorylation and nuclear translocation of IRF-7, ORF45 efficiently inhibits the activation of interferon alpha and beta genes during viral infection. Inhibition of interferon gene expression through a viral protein blocking the activation and nuclear translocation of a crucial transcription factor is a novel mechanism for viral immune evasion. PMID- 11943873 TI - Cooperative anion binding and electrochemical sensing by modular podands. AB - A series of podands based on two or three hydrogen bonding "arms" situated in mutually ortho, meta, or para relationships about an aryl core have been prepared, and their affinities for simple inorganic anions were measured. Of the two-arm hosts the meta compound and to a lesser extent the ortho host exhibit a cooperative anion binding effect. The two arms function essentially independently in the para derivative. The mutually meta three-arm host shows dramatically enhanced cooperative binding. Conformational changes within the meta two-arm host result in significantly enhanced electrochemical anion sensing compared with the more conformationally rigid three-arm host. PMID- 11943874 TI - Probing alkali metal-pi interactions with the side chain residue of tryptophan. AB - Feeble forces play a significant role in the organization of proteins. These include hydrogen bonding, hydrophobic interactions, salt bridge formation, and steric interactions. The alkali metal cation-pi interaction is a force of potentially profound importance but its consideration in biology has been limited by the lack of experimental evidence. Our previous studies of cation-pi interactions with Na(+) and K(+) involved the side arms of tryptophan (indole), tyrosine (phenol), and phenylalanine (benzene) as the arene donors. The receptor system possesses limiting steric constraints. In this report, we show that direct interactions between alkali metals and arenes occur at or within the van der Waals contact distance. PMID- 11943872 TI - Neuroprotection in cerebral ischemia by neutralization of 3-aminopropanal. AB - Cerebral ischemia stimulates increased activity of polyamine oxidase, a ubiquitous enzyme that catabolizes polyamines to produce 3-aminopropanal. 3 Aminopropanal is a reactive aldehyde that mediates progressive neuronal necrosis and glial apoptosis. Here we report that increased levels of 3-aminopropanal modified protein levels in humans after aneurysmal subarachnoid hemorrhage correlate with the degree of cerebral injury as measured by admission Hunt/Hess grade. In vitro screening of clinically approved drugs reveals that N-2 mercaptopropionyl glycine (N-2-MPG), an agent clinically approved for prevention of renal stones in patients with cysteinuria, significantly inhibits the cytotoxicity of 3-aminopropanal. N-2-MPG reacts with 3-aminopropanal to yield a nontoxic thioacetal adduct, as confirmed by electrospray ionization mass spectroscopy. Administration of N-2-MPG in clinically relevant doses to rats significantly reduces cerebral 3-aminopropanal-modified protein immunoreactivity and infarct volume in a standardized model of middle cerebral artery occlusion, even when the agent is administered after the onset of ischemia. These results implicate 3-aminopropanal as a therapeutic target for cerebral ischemia. PMID- 11943875 TI - Organization of the interior of molecular capsules by hydrogen bonding. AB - The enclosure of functional entities within a protective boundary is an essential feature of biological systems. On a molecular scale, free-standing capsules with an internal volume sufficiently large to house molecular species have been synthesized and studied for more than a decade. These capsules have been prepared by either covalent synthesis or self-assembly, and the internal volumes have ranged from 200 to 1,500 A(3). Although biological systems possess a remarkable degree of order within the protective boundaries, to date only steric constraints have been used to order the guests within molecular capsules. In this article we describe the synthesis and characterization of hexameric molecular capsules held together by hydrogen bonding. These capsules possess internal order of the guests brought about by hydrogen bond donors within, but not used by, the framework of the capsule. The basic building blocks of the hexameric capsules are tetrameric macrocycles related to resorcin[4]arenes and pyrogallol[4]arenes. The former contain four 1,3-dihydroxybenzene rings bridged together by -CHR- units, whereas the latter contain four 1,2,3-trihydroxybenzene rings bridged together. We now report the synthesis of related mixed macrocycles, and the main focus is on the macrocycle composed of three 1,2,3-trihydroxybenzene rings and one 1,3 dihydroxybenzene ring bridged together. The mixed macrocycles self-assemble from a mixture of closely related compounds to form the hexameric capsule with internally ordered guests. PMID- 11943876 TI - Calcium release and influx in yeast: TRPC and VGCC rule another kingdom. AB - Calcium is one of the most ubiquitous second messengers, in addition to being a fundamentally important cofactor for many proteins' functions. Thus, cells from fungi to plants to animals have evolved mechanisms for carefully controlling calcium concentrations in their organelles and cytosol. This perspective discusses the similarities and differences between yeast and animals in cation channels of two families: the transient receptor potential channels (TRPCs) and voltage-gated calcium channels (VGCCs). One of the key differences highlighted is in the channels implicated in the response of yeast and animal cells to depletion of calcium from intracellular stores, and the differences in localization of these channels between yeast and animal cells. Understanding the logic of fungal Ca(2+) channels, therefore, may provide new insights into the organization and regulation of cellular calcium signaling networks in animals. PMID- 11943877 TI - ADAP-ting TCR signaling to integrins. AB - Adaptor proteins are essential components of T cell receptor (TCR) signaling cascades regulating gene transcription and cytoskeletal reorganization. The molecular adaptor adhesion- and degranulation-promoting adaptor protein (ADAP), also known as Fyn binding protein (FYB) or Slp-76-associated protein of 130 kilodaltons (SLAP-130), interacts with a number of signaling intermediates including Slp-76, the Src family tyrosine kinase Fyn, vasodilator-stimulated phosphoprotein (VASP), and the actin-nucleating protein WASP. Recently ADAP was shown genetically to positively regulate T cell activation, TCR-induced integrin clustering, and T cell adhesion. The mechanism by which ADAP couples TCR stimulation to integrin clustering remains unclear; however, studies of ADAP, the exchange factor Vav1, and WASP suggest that TCR and integrin clustering may be controlled by distinct signaling pathways. PMID- 11943878 TI - Integration of signals from receptor tyrosine kinases and g protein-coupled receptors. AB - Activation of G protein-coupled receptors (GPCRs) leads to stimulation of classical G protein signaling pathways. In addition, GPCRs can activate the mitogen-activated protein kinases (MAPKs) such as the extracellular signal regulated kinases, c-Jun NH(2)-terminal kinases (JNKs), and p38 MAPKs, and thereby influence cell proliferation, cell differentiation and mitogenesis. Cross talk between GPCRs and receptor tyrosine kinases (RTKs) is an incredibly complex process, and the exact signaling molecules involved are largely dependent on the cell type and the type of receptor that is activated. In this review we investigate recent advances that have been made in understanding the mechanisms of cross talk between GPCRs and RTKs, with a focus on GPCR-mediated activation of the Ras/MAPK pathway, GPCR-induced transactivation of RTKs, GPCR-mediated activation of JNK, and p38 MAPK, integration of signals by RhoGTPases, and activation of G protein signaling pathways by RTKs. PMID- 11943879 TI - Prostanoid signal integration and cross talk. AB - The enzymatic machinery for the production of prostanoids and the receptors responsible for detecting their presence are widely distributed in the body. One pair of prostanoids, prostacyclin and thromboxane A(2), are particularly important in the control of haemodynamics and haemostasis. Prostacyclin achieves its antiplatelet effect by acting as a physiological antagonist, but displays some selectivity towards thromboxane A(2)-mediated platelet activation, possibly by virtue of the inability of thromboxane A(2) receptors to couple directly to G(i) proteins, and because platelet-derived endoperoxides can act as substrates for prostacyclin synthesis in endothelial cells. At low concentrations, prostaglandin E(2) can synergize with thromboxane A(2) by acting on the EP(3) subtype of prostaglandin E(2) receptor, resulting in opposition to the protective function of prostacyclin. In contrast, high concentrations of prostaglandin E(2) act on the prostacyclin receptor, and possibly the prostaglandin D(2) receptor, to turn off platelet activation. Integration of prostanoid signalling in the vascular system is similarly complex, and interpretation of data is further complicated by the regional distribution of prostanoid receptors in different vascular beds, and the poor selectivity of agonists and antagonists. PMID- 11943880 TI - Ligand rescue of constitutively active mutant receptors. AB - Constitutively active mutants (CAMs) of G protein-coupled receptors are found naturally in disease states and they can be generated by point mutation. As these mutants are able to activate G proteins in the absence of a ligand, they are useful tools in the study of conformational changes leading to receptor activation and in the drug discovery process. Early studies on CAMs noted that they are often expressed at lower levels than their wild-type forms. In this review we discuss the mechanisms responsible for this reduced expression and also provide an explanation for the observation that challenging cells with receptor ligands can increase the CAM expression level. The application of these observations to the development of a high-throughput reporter assay suitable for ligand identification is also discussed. PMID- 11943882 TI - Mechanisms of cross-talk between G-protein-coupled receptors. AB - There is increasing evidence to suggest that 'cross-talk' occurs between G protein-coupled receptors and their intracellular second messenger pathways. Cross-talk between different pathways may occur at the level of receptors, G proteins, effectors or second messengers and may serve to fine-tune cell signalling. There is a growing body of evidence to suggest that cellular compartmentalization may play a crucial role in regulating these cross-talk interactions. Understanding the mechanisms of cross-talk may therefore be the key to the design and application of future therapeutics and the development of drug specificity. PMID- 11943883 TI - The signaling pathways mediated by P2Y nucleotide receptors in the formation and maintenance of the skeletal neuromuscular junction. AB - The motor neuron, the Schwann cell and the muscle cell are highly specialized at the vertebrate skeletal neuromuscular junction (NMJ). The muscle cell surface contains a high local density of acetylcholine (ACh) receptors (AChRs), acetylcholinesterase (AChE) and their interacting macromolecules at the NMJ, forming the postsynaptic specializations. During the early stages of development, the incoming nerve terminal induces the formation of these postsynaptic specializations; the nerve secretes agrin and neuregulin (NRG), which are known to aggregate existing AChRs and to increase the expression of AChR at the synaptic region, respectively. In addition, adenosine 5'-triphosphate (ATP) is stored at the motor nerve terminals and is coreleased with ACh during muscle contraction. Recent evidence suggests that ATP can play a role in forming and maintaining the postsynaptic specializations by activating its corresponding receptors. In particular, one of the nucleotide receptor subtypes, the P2Y(1) receptor, is specifically localized at the NMJs. The gene expression of AChR and AChE is upregulated after the activation of P2Y(1) receptors. Thus, the synaptic ATP together with agrin and NRG can act as a synapse-organizing factor to induce the expression of postsynaptic functional effectors. PMID- 11943881 TI - Diversity of G protein-coupled receptor signaling pathways to ERK/MAP kinase. AB - One of the most intriguing examples of cross talk between signaling systems is the interrelationship between G protein-coupled receptor and growth factor receptor pathways leading to activation of the ERK/MAP kinase phosphorylation cascade. This review focuses on the mechanism of this cross talk, denoting primarily signaling components known to occur in the G protein-coupled receptor branch of the MAP kinase pathways in neural cells. Recent evidence is presented on the existence of a plethora of pathways, due to the multiplicity of G protein coupled receptors, their differential interaction with heterotrimeric G protein isoforms, various effectors and second messengers. In light of this rich diversity, the review will discuss different points of convergence of G protein coupled receptor and growth factor receptor pathways that may feature a requirement for growth factor receptor transactivation, receptor internalization and scaffolds to assemble receptor, adaptor and anchoring proteins into multiprotein complexes. PMID- 11943885 TI - Role of neck surgery in conjunction with radiation in regional control of node positive cancer of the oropharynx. AB - For patients with clinically node-positive oropharynx cancer treated with radiotherapy, planned neck dissection is controversial. We investigated whether neck surgery after radiation reduces nodal recurrence. Between 1970 and 1995, 263 patients at Massachusetts General Hospital received radiotherapy for clinically node-positive base of tongue or tonsil carcinomas. Patients received three different types of treatment: neck surgery followed by radiation (SR)-50 patients; radiation alone (RT)-160 patients; and radiation followed by surgery (RS)-53 patients. Median patient follow-up was 28 months. SR patients have an 84% complete response (CR) rate, RT patients 76%, and RS patients 13%. In multivariate analysis, among patients with a CR, the three treatment groups have the same regional control rates. Among patients with an incomplete response, the RS treatment group is 67% (p < 0.01) and 86% (p < 0.01) less likely to have recurrence than the RT and SR groups, respectively. Neck dissection after radiation therapy improves regional control for patients without a complete clinical response to radiation therapy but not for those with a CR. Despite higher CR rates, neck dissection before radiation confers no regional control benefit. We therefore recommend that primary radiotherapy with neck dissection be reserved for those without a complete clinical response. PMID- 11943886 TI - Intracranial ependymomas: an analysis of prognostic factors and patterns of failure. AB - From 1965 to 1997, 49 patients were diagnosed and treated for intracranial ependymoma at one institution. Tumor location was infratentorial in two thirds, and pathology was low grade in 38 patients (78%). Gross total resection of the primary tumor was achieved in 21 (43%). Thirty-six patients received adjuvant radiotherapy; the entire neuraxis was treated in 14, whole brain in 10, and local field only in 12. Median follow-up was 9.6 years (range, 2-33 years). The 5-, 10 , and 15-year overall survival rates were 71.4%, 63.5%, and 63.5% for craniospinal radiotherapy, 60.0%, 60.0%, and 40.0% for whole brain radiotherapy, and 80.8%, 64.6%, and 64.6% for local field radiotherapy (p = 0.88). The 5-, 10-, and 15-year local control rates were 60.3%, 54.4%, and 48.9%. The prognostic factors for a better local control rate were gross total resection (p = 0.021) and low grade histology (p = 0.031). Seventeen of 43 (39.5%) M0 patients did not respond to treatment; all had local failure and 4 also had a spinal relapse. Spinal relapse developed in 3 of 31 (10%) M0 patients who did not receive spinal radiotherapy, whereas 1 of 12 (8%) who had spinal radiotherapy did not respond to treatment in the spine. The results of this study indicate that local radiotherapy is sufficient for M0 patients with intracranial ependymoma. PMID- 11943888 TI - Palliative 5-fluorouracil-based chemotherapy for advanced colorectal cancer in the elderly: results of a 10-year experience. AB - The aim of this study was to evaluate the objective tumor response rates and toxicities in elderly patients (older than 70 years) with advanced colorectal cancer treated with 5-fluorouracil (5-FU) as a first-line palliative chemotherapy regimen. Eighty-six consecutive patients were enrolled onto a retrospective study between January 1990 and February 1998. Patients were divided into two groups; group 1 consisted of patients who were 70 to 74 years old, and group 2 consisted of patients who were age 75 years and older. Four types of 5-FU-based chemotherapy were administered. First-line chemotherapy was continued until disease progression, unacceptable toxicity, or patient refusal. Primary tumor sites were the colon (n = 44), rectum (n = 29), and colorectal (n = 13). There was no difference in response (complete/partial) rates according to age groups (group 1: 21%, group 2: 26%). Median overall survival was 17 months for group 1 and 14 months for group 2, with 1-year survival at 63% and 55%, respectively (not statistically significant). Median time to progression was 6 months for both age groups. Chemotherapy in both groups increased performance status, and weight in 26% and 31%, respectively. No toxic death was reported. There was no difference in overall or severe toxicity between the two age groups, and all adverse events were manageable. Toxicity (grade III/IV) occurred in 25% of patients. Based on these findings, palliative first-line chemotherapy for colorectal cancer can be performed in selected elderly patients without increasing either morbidity or mortality, while obtaining response rates and side effects comparable to those in younger patients. PMID- 11943887 TI - Low activity of 13-cis-retinoic acid plus interferon alpha 2a in advanced renal cell carcinoma. AB - Retinoids and interferon alpha have shown synergistic activity against metastatic renal cell carcinoma in previous preclinical and clinical studies. Based on these results, we conducted a phase II trial of 13-cis-retinoic acid (cRA) at 1 mg/kg/dose interferon alpha2a (IFN) at initial dose of 9 MU three times a week. Thirty-one patients were entered, all evaluable for toxicity and 30 evaluable for response. One patient achieved a partial response and 10 patients achieved stable disease. Toxicity was mild and primarily related to interferon. No toxic deaths were reported. Median survival time was 10 months. At the dose and schedule used, cRA and interferon-alpha2a showed low activity against metastatic renal cell carcinoma. Further studies with this combination are not recommended. PMID- 11943889 TI - Influence of gamma knife radiosurgery on the quality of life in patients with brain metastases. AB - Quality of life (QOL) is an important issue in the treatment of patients with brain metastases. With median survival times often less than 4 months, less invasive treatment options that maximize QOL parameters are essential. In recent years, stereotactic radiosurgery (SRS) has been commonly used as a noninvasive alternative to surgical resection for such patients. This prospective study was undertaken to evaluate QOL in patients undergoing SRS for brain metastases. Between 1999 and 2000, 20 patients with metastatic disease to the brain were evaluated and treated in our Gamma Knife unit. All patients performed the Spitzer QOL survey (10-point scale) both before stereotactic radiosurgery and at each follow-up visit. Primary sites of disease included lung (n = 10), breast (n = 5), melanoma (n = 2), thyroid (n = 1), uterine (n = 1), and kidney (n = 1). Fifteen (75%) had prior whole brain radiotherapy (median dose: 35 Gy). The median age and Karnofsky Performance Status were 58 years and 80, respectively. The median Spitzer score before SRS was 9 (range: 7-10), and the median follow-up time of the patients in this series was 7 months. The median posttreatment Spitzer score at 1 and 3 months after SRS was 9 (range: 5-10) and 8 (range: 4-10), respectively. Crude intracranial tumor control in this cohort of patients was 90%. Extracranial tumor progression was noted in 8 patients (40%), and in these patients, Spitzer scores tended to decrease in value. In those patients who had no evidence of intracranial or extracranial tumor progression, Spitzer scores remained either unchanged or improved. Gamma knife SRS is an appropriate treatment modality for maintaining QOL parameters in patients with brain metastases. Tumor progression both intracranially and extracranially influences QOL parameters. Confirmation of this finding will require further investigation. PMID- 11943890 TI - Primary intraspinal primitive neuroectodermal tumor: case report of a tumor arising from the sacral spinal nerve root and review of the literature. AB - Primary spinal primitive neuroectodermal tumor (PNET) is a rare condition, 18 cases of which have been reported in the literature. In general, this tumor is treated with surgery followed by radiotherapy and chemotherapy, but prognosis is still poor. An 18-year-old female patient with an intradural, extramedullary mass at L3-L5 levels is presented in this report. This is the first female patient with primary spinal PNET at lumbar region, second patient with spinal nerve root origin, and third one with intradural, extramedullary localization ever reported in the literature. After surgery, she was treated with craniospinal radiotherapy and four cycles of combination chemotherapy regimen consisting of vincristine, cyclophosphamide, doxorubicin alternated with ifosfamide, and VP-16. Currently, she is asymptomatic and alive at 25 months. The histopathologic, radiologic, and clinical findings of the patient are presented and relevant literature is reviewed. PMID- 11943891 TI - Predictors of short-term survival and progression to chemotherapy in patients with advanced colorectal cancer treated with 5-fluorouracil-based regimens. AB - The aim of this study was to assess in patients with advanced colorectal cancer which factors were associated with short-term survival (6 months or less) and progression to first-line 5-fluorouracil (5-FU) chemotherapy. Three hundred twenty-one consecutive nonselected patients with advanced colorectal cancer were treated with conventional 5-FU-based regimens as first-line treatment from 1988 to 1999. Factors related to patient, tumor, or treatment were analyzed by univariate and multivariate logistic regression analysis by comparing short survivors (SS, those who survived or= 2) (p = 0.015), elevated (>or=5 microg/l) serum carcinoembryonic antigen (CEA) (p = 0.015), and more than one site of metastatic disease (p < 0.001). Progression to first-line chemotherapy (p < 0.001) was also a strong factor associated with short survival in multivariate analysis; factors predictive of progression were elevated CEA (p = 0.027) and diffuse metastatic disease (p = 0.029). Our data indicate the relevance of some clinical prognostic factors (younger age, poor performance status, elevated CEA, site of primary, number of metastatic sites, resistance to chemotherapy) as independent factors associated with poor survival and progression to first-line chemotherapy in patients with metastatic colorectal cancer treated with conventional 5-FU regimens. Patients identified by these factors as having a poor prognosis and low probability of response to treatment should be considered either for more aggressive regimens or supportive care only: conventional 5-FU treatments do not impact on response or survival. PMID- 11943892 TI - Radiotherapy alone in the treatment of clinical stage I-IIA, nonbulky, Hodgkin's disease: single-institution experience on 73 patients staged with lymphangiography and laparoscopy. AB - From 1985 to 1998, at the Regional Cancer Center of Padua, patients with Hodgkin's disease (HD) routinely underwent a clinical staging procedure including lymphangiography and laparoscopy with multiple liver and spleen biopsies. Patients with IA and IIA nonbulky HD were treated with radiotherapy alone. The aim of this study is to analyze the efficacy of radiotherapy as radical treatment in this group of patients, and the role of lymphangiography and laparoscopy in the selection of patients with abdominal disease located to the spleen, liver, or the pelvic lymphatic chains. From January 1985 to January 1998, 94 previously untreated patients with biopsy-proven HD underwent clinical staging procedures consisting of history, physical examination, routine laboratory tests, chest radiography, total-body computed tomography scan, and bone marrow biopsy and were considered in stage I-IIA nonbulky. In addition, all patients underwent bipedal lymphangiography, which was positive in 12 (12.8%). Of the 82 patients with negative lymphangiography, 9 (11%) showed disease below the diaphragm at laparoscopy with multiple random spleen and liver biopsies. Of the remaining 73 patients, 32 were male and 41 were female with a median age of 29 years (range: 14-72 years). PMID- 11943894 TI - Poorly differentiated colon carcinoma with neuroendocrine features presenting with hypercalcemia and cutaneous metastases: case report and review of the literature. AB - Humoral hypercalcemia is rarely associated with colon carcinoma; cutaneous metastases from colon carcinoma are also infrequent. To the authors' knowledge, no cases of colon carcinoma presenting with both hypercalcemia and cutaneous metastases have been reported to date. A case of advanced poorly differentiated colon carcinoma with neuroendocrine features with both humoral hypercalcemia of malignancy (HHM) and cutaneous metastases is presented. A poorly differentiated colon carcinoma with neuroendocrine features occurred in a 42-year-old patient with metastases to the liver, both femurs, left orbit, and scalp. The hypercalcemia was caused by the expression of a parathyroid hormone related peptide by both the primary and cutaneous metastatic tumors. Bisphosphonate treatment helped normalize serum calcium in a few days, but hypercalcemia recurred approximately 3 weeks later. Chemotherapy only mildly reduced the size of the cutaneous metastases. The patient died 8 months after initial diagnosis. To the authors' knowledge, the case presented in the current study is the first to be reported with both HHM and cutaneous metastases. Hypercalcemia and cutaneous metastases are separately associated with a poor prognosis and indicate advanced and widely metastatic disease. Although still unclear, the mechanism by which colon cancer causes cutaneous metastases and hypercalcemia, in light of current theories presented in the literature, is discussed. PMID- 11943893 TI - Phase I/II trial of outpatient docetaxel, cisplatin, 5-fluorouracil, leucovorin (opTPFL) as induction for squamous cell carcinoma of the head and neck (SCCHN). AB - The purpose of this study was to establish the maximum tolerated dose (MTD) of docetaxel in an outpatient docetaxel (T), cisplatin (P), 5-fluorouracil (5-FU) (F), and leucovorin (L) (opTPFL) regimen and to obtain preliminary assessment of opTPFL efficacy. Thirty-four patients with stage III or IV squamous cell carcinoma of the head and neck were treated with opTPFL. Docetaxel was escalated from 60 to 95 mg/m(2) in combination with 100 mg/m(2) cisplatin intravenous bolus, and 2,800 mg/m(2) 5-FU continuous infusion and 2,000 mg/m(2) leucovorin continuous infusion with prophylactic growth factors and antibiotics. Patients who achieved a complete (CR) or partial (PR) response to three cycles received definitive twice-daily radiation therapy. A total of 97 cycles were administered to 34 patients. The major acute toxicities were neutropenia and mucositis. The MTD of docetaxel was 90 mg/m(2) . Seventy-seven of 97 cycles of were administered on an outpatient basis. The overall clinical response rate to opTPFL was 94%, with 44% CRs and 50% PRs. The MTD of opTPFL is 90 mg/m(2) docetaxel. Outpatient administration of opTPFL is tolerable, feasible, and does not alter the ability to administer definitive radiation therapy on schedule. PMID- 11943896 TI - Osteoradionecrosis of the mandible: scientific basis for clinical staging. AB - Osteoradionecrosis of the mandible can be a serious complication of radiotherapy for head and neck cancer. The disorder generally presents in one of several distinct clinical variations. This is probably because of a complex interaction of etiologic factors. A clinical staging system for the disorder is described. Such a system is necessary for the conduct of clinical research, and for the development and assessment of treatment regimens. PMID- 11943895 TI - Prognostic role of nm23 gene expression in patients with ovarian cancer. AB - The aim of the study was to define the prognostic role of the metastasis suppressor gene, nm23, in 50 patients with primary ovarian cancer. Immunohistochemical analysis was performed on formalin-fixed, paraffin-embedded specimens by the primary nm23 monoclonal antibody (Novocastra, NCL-nm23 clone 37.6). Forty-two specimens (84%) showed a positive nm23 staining. The nm23 staining was more intensive in patients with normal serum CA19.9 levels, patients with nonrecurrent disease, and alive patients (p < 0.05). Nm23 expression did not correlate with common clinicopathologic parameters such as histology, grade of differentiation, International Federation of Gynecology and Obstetrics stage, and CA-125. Although the difference was not statistically significant (p = 0.11), we found that nm23 may have a favorable prognostic factor in ovarian cancer. To clarify this subject further, prospective studies on a larger population are needed. PMID- 11943898 TI - Chromosomal aberrations in intravascular lymphomatosis. AB - Intravascular lymphomatosis (IVL) is a unique, disseminated type of malignant lymphoma. However, no detailed comparative study limited to the chromosomal aberrations of IVL has been reported, because IVL is extremely rare and difficult to diagnose while the patient is alive. We present here a case of IVL, and compare its karyotype with those of five cases of previously reported IVL. The accumulation of structural aberrations in chromosomes 1, 6, and 18, especially 1p (4 of 6 cases) and trisomy 18 (4 of 6 cases), were found in our comparative study of the B-cell lineage typical IVL. These chromosomal rearrangements must provide important information regarding the characteristics of cytogenetically associated with the cellular genetics of IVL. PMID- 11943897 TI - Clinical outcome and prognosis of patients with inflammatory breast cancer. AB - This report analyzes clinical factors affecting outcome in 26 patients with inflammatory breast cancer. Peau d'orange was the most common clinical finding at diagnosis (65%). A palpable breast mass (PBM) was noted in 65% with axillary lymph node involvement in 81% of patients. Eighteen patients were staged as stage IIIB and eight as stage IV. Initial metastases included supraclavicular nodes (five of eight), bones (one of eight), skin (one of eight), and liver (one of eight). All patients were treated with neoadjuvant chemotherapy (cyclophosphamide, doxorubicin, and fluorouracil, 18 patients; other, 8 patients). Partial response was the best clinical response attained in 38% of patients. Only one patient was treated with total mastectomy after neoadjuvant chemotherapy, and 19 patients received radiotherapy followed (2 patients) or not (17 patients) by mastectomy. The progression rate in stage IIIB patients was 78%, with distant sites of progression in 93% of patients and only 7% with local progression. Mean time-to-progression was 13 months (Kaplan-Meier estimates of 45% and 11% at 24 and 48 months, respectively). The median overall survival (OS) value of the entire population was 13.2 months (Kaplan-Meier estimates at 24 and 48 months of 21% and 12.5%). By Kaplan-Meier method and log-rank test, a better OS was correlated with stage IIIB (p = 0.002), a PBM at diagnosis (p = 0.01), and a favorable response to initial chemotherapy (p = 0.03). Our results confirm the better clinical outcome of patients with stage IIIB and PBM at diagnosis. They also support the role for combined treatment as the best modality approach for this disease. However, overall prognosis remained poor, with recurrence and death resulting from the disease. PMID- 11943899 TI - Prognostic factors for patients with metastatic colorectal cancer receiving protracted venous infusion of 5-FU. AB - The objective of this study was to investigate the possible prognostic factors in patients with metastatic colorectal cancer (CRC) treated with protracted venous infusion (PVI) of 5-fluorouracil (5-FU). PVI of 5-FU is considered to be one of the standard therapies for patients with CRC because of its high efficacy and low toxicity, but possible prognostic factors in patients with CRC treated with PVI of 5-FU have not been reported until now. One hundred two consecutive patients with metastatic CRC, treated with PVI of 250 mg/m(2) /d of 5-FU at the National Cancer Center Hospital between January 1991 and December 1998, were investigated retrospectively. The data were evaluated for possible correlations with clinicopathologic indices or prognosis. In this study, grade III nausea, stomatitis, diarrhea, and hand-foot syndrome occurred in only 3%, 5%, 5%, and 6% of cases, respectively. No grade III or more hematologic toxicities were observed. The overall response (complete response + partial response) rate for all 102 cases was 26%, with a 95% CI of 18% to 35%. Overall median survival time for the 102 cases was 303 days. In multivariate analysis, the favorable prognostic factors were normal albumin level ( 700% as CaCl2 increased from 0 to 5 mmol/l. Above 5 mmol/l, no additional increases occurred. With batroxobin, PCF did not develop at CaCl2 concentrations < or = 2.5 mmol/l. Above 2.5 mmol/l CaCl2, PCF values increased and at 10 mmol/l CaCl2 were equal to those seen with thrombin. CEM in batroxobin-mediated clots peaked at 10 mmol/l CaCl2 but were 40% less than the values found in thrombin-mediated clots. When the thrombin inhibitor P-PACK was added to the batroxobin system, dose-dependent decreases in PCF and CEM were noted. At 120 micromol/l, P-PACK totally suppressed PCF. PCF in blood from a factor VIII-deficient patient varied significantly when clotted with batroxobin versus thrombin. PCF development in factor VIII-deficient blood was normal with thrombin but is delayed and depressed with batroxobin. PCF values in factor VIII deficient blood did not reach the thrombin value after 1200 s of clotting, and CEM was significantly less. These results confirm that PCF development is thrombin dependent and that delay or reduction of PCF development results in structurally weaker clots. PMID- 11943934 TI - The activated protein C (APC)-resistant phenotype of APC cleavage site mutants of recombinant factor V in a reconstituted plasma model. AB - Recently, new missense mutations in the activated protein C (APC) cleavage sites of human factor V (FV) distinct from the R506Q (FV Leiden) mutation have been reported. These mutations affect the APC cleavage site at arginine (Arg) 306 in the heavy chain of activated FV. Whether these mutations result in APC resistance and are associated with a risk of thrombosis is not clear. The main objective of the present study was to identify the APC-resistant phenotype of FV molecules with different mutations in APC cleavage sites. To study this, recombinant FV mutants were reconstituted in FV-deficient plasma, after which normalized APC sensitivity ratios (n-APC-SRs) were measured in activated partial thromboplastin time-based and Russell's Viper Venom time-based APC-resistance tests. The mutations introduced in FV were R306G, R306T, R506Q, R679A and combinations of these mutations. Based on the APC-sensitivity ratios, we conclude that the naturally occurring mutations at Arg306 (i.e. FV HongKong and FV Cambridge) result in a mildly reduced sensitivity for APC (n-APC-SR, 0.74-0.87), whereas much lower values (n-APC-SR, 0.41-0.51) are obtained for the mutation at Arg506 (FV Leiden). No effect on the n-APC-SR was observed for the recombinant FV mutant containing the single Ala679 mutation. Because reduced sensitivity for APC, not due to FV Leiden, is a risk factor for venous thrombosis, these data suggest that mutations at Arg306 might be associated with a mild risk of venous thrombosis. PMID- 11943933 TI - The use of coagulation activation markers (soluble fibrin polymer, TpP, prothrombin fragment 1.2, thrombin-antithrombin, and D-dimer) in the assessment of hypercoagulability in patients with inherited and acquired prothrombotic disorders. AB - A total of 260 consecutive patients, referred for hypercoagulable assessment, was included in this study. Four coagulation activation markers were utilized to assess these patients [enzyme-linked immunosorbent assays for soluble fibrin polymer (TpP), prothrombin fragment 1.2, thrombin-antithrombin complex, and D dimer]. The mean levels of the activation markers directly correlated with the number of hypercoagulable abnormalities. The percentage of patients with increased TpP levels for each group was lower than the other activation markers. The findings indicate that activation markers reflect the number of underlying thrombophilic abnormalities. Our data suggest that there is a utility in performing a panel of coagulation activation markers to assess the thrombotic risk. The measurement of soluble fibrin polymer may be more reflective of an impending vascular event. PMID- 11943935 TI - Genotype associations of factor VII gene with plasma factor VII coagulant activity and antigen levels in healthy Chinese. AB - A raised plasma factor VII (FVII) level is one of the risk factors for coronary artery disease. The R353Q polymorphism at codon 353 and the 10 base pair (bp) insertion (0/10 bp) polymorphism of the FVII gene have been reported to be associated with plasma FVII levels in several populations. We investigated these two polymorphisms in 209 male and 214 female healthy Chinese. The allele frequencies of 10 bp and Q were 0.036 and 0.045, respectively. Strong linkage disequilibrium was observed between these two sites (Delta = 0.85, P < 0.001). There were significant genotype associations of these two loci with FVII coagulant activity (FVIIc) and antigen (FVIIAg) levels. Heterozygous individuals had lower FVIIc and FVIIAg levels than those homozygous for the common alleles. When analyzed separately by gender, the 0/10 bp polymorphism was strongly associated with FVIIAg levels in males and females. However, both polymorphisms were significantly associated with FVIIc levels only in the females. The effect of 0/10 bp polymorphism predominated over that of the R353Q polymorphism in a two way analysis of variance procedure. In the Chinese, the 10 bp insertion may reduce transcription of the FVII gene, leading to the decreased synthesis of FVII protein and thus FVIIc. PMID- 11943936 TI - Desmopressin in the treatment of menorrhagia in women with no common coagulation factor deficiency but with prolonged bleeding time. AB - The objective of this study was to investigate the efficacy and safety of desmopressin (1-desamino-8-D-arginine vasopressin) compared with placebo in the reduction of menstrual blood loss in women with menorrhagia and prolonged bleeding time, but without common coagulation factor deficiencies. We performed a randomized, double-blind, cross-over study using 300 microg desmopressin nasal inhalation or placebo treatment in one of the two first treatment cycles. Desmopressin was given only for the 2 days during which the bleeding had been at a maximum in the previous baseline cycle. A third open cycle involved combined treatment with desmopressin and tranexamic acid during the 2 days for all patients. Menstrual blood loss during the treatment periods was compared with blood loss during placebo-treated periods using objective measurement. A significant reduction of menstrual blood loss was found in the cycles treated with combined desmopressin and tranexamic acid compared with placebo. When analyzing the blood loss during the two treatment days, there was a significant reduction in blood loss for the 2 days with desmopressin alone versus placebo. The treatment was well tolerated and no serious adverse events were recorded. In conclusion, we find that nasal desmopressin is a possible complement for the medical treatment of menorrhagia. PMID- 11943937 TI - Beneficial effect of JTV-803, a new synthetic inhibitor of activated factor X, against both lipopolysaccharide-induced and tissue factor-induced disseminated intravascular coagulation in rat models. AB - We examined whether JTV-803, a specific activated factor X inhibitor independent of antithrombin III (ATIII), is effective against disseminated intravascular coagulation (DIC) in rat models induced by tissue factor (TF) or lipopolysaccharides (LPS). In male Wistar rats, DIC was induced by a 4 h infusion of thromboplastin (3.75 U/kg) or LPS (50 mg/kg). The rats were given JTV-803 (0.3 or 3 mg/kg, bolus intravenously) (JTV-803 groups) or low molecular weight heparin (LMWH groups) (200 U/kg, bolus intravenously) prior to an injection of TF or LPS. The results showed that JTV-803 was dose-dependently effective against DIC in both TF-induced and LPS-induced rat models. This anti-DIC effect of JTV-803 at higher doses was almost equivalent to that of LMWH in both types of DIC. Plasma ATIII activity was more prominent in the group treated with JTV-803 than in that treated with LMWH. None of rats died in the TF-induced DIC model with or without drug administration. On the contrary, seven of 22 rats died (mortality rate, 31.8%) in the LPS-induced DIC model without drug administration. Although the mortality rate of rats induced with LPS and treated with LMWH was quite high (6/16, 37.5%), none of the LPS-induced rats treated with JTV-803 died. These findings suggested that JTV-803 can treat both TF-induced and LPS-induced DIC models, and that this drug has greater potential in preserving ATIII and in improving the prognosis of DIC. PMID- 11943938 TI - Decrease in sensitivity of D-dimer for acute venous thromboembolism after starting anticoagulant therapy. AB - D-dimer testing is useful for the exclusion of acute venous thromboembolism (VTE). Anticoagulant therapy is expected to reduce D-dimer levels in patients with thrombosis and, consequently, it may not be safe to use D-dimer levels to exclude VTE after anticoagulant therapy has been started. The objectives of this study were to estimate the decrease in D-dimer levels after 24 h of heparin therapy and, applying this estimate to the results of a recent study, to calculate the expected reduction in sensitivity. Using pre-defined criteria, we first performed a literature review to determine whether, and by how much, D dimer levels decrease within 24 h of starting heparin therapy in patients with acute VTE. Using D-dimer levels that were measured in a prospective study of patients with confirmed deep vein thrombosis and/or pulmonary embolism as baselines, we then determined the change in sensitivity (and specificity) that would result from the fall in D-dimer levels that the literature review suggested would have occurred after 24 h of heparin therapy. On the basis of the literature review, we calculated that mean D-dimer levels decrease by 25%, 24 h after starting heparin therapy in patients with acute VTE. This 25% decrease in D-dimer levels resulted in a decrease in sensitivity from 95.6% (95% confidence interval, 90.0-98.6) to 89.4% (95% confidence interval, 83.7-95.1). There is a decrease in D-dimer levels in patients with acute VTE 24 h after starting heparin therapy that is expected to result in a clinically important drop in sensitivity. PMID- 11943940 TI - Massive choroidal hemorrhage associated with low molecular weight heparin therapy. AB - An 84-year-old woman with unstable angina pectoris was treated with subcutaneous enoxaparine (Clexane) for several days before presenting with severe pain and decreased vision in her left eye. The intraocular pressure was 70 mmHg, and fundus examination showed a pigmented choroidal lesion and associated choroidal and retinal detachment. Ultrasonography was consistent with choroidal hemorrhage, and she was diagnosed as having acute glaucoma secondary to massive subchoroidal hemorrhage. Medical control of the intraocular pressure resulted in a significant clinical improvement. Intraocular hemorrhage and angle-closure glaucoma are rare and previously unreported complications in patients treated with low molecular weight heparin. It is important to be aware of this ocular complication as these drugs are so often used. PMID- 11943939 TI - Coagulation indicators in chronic stable effort angina and unstable angina: relationship with acute phase reactants and clinical outcome. AB - The aim of the study was to evaluate which pattern of coagulation indicators characterizes unstable angina and, particularly, its relationship with short-term prognosis. Forty patients with unstable angina (UA Group) at admission in the intensive care unit, 40 patients with chronic stable effort angina (SEA Group), and 20 age- and sex-matched healthy controls were studied. Blood coagulation indicators were fibrinogen, prothrombin fragment F1 + 2 (F1 + 2), thrombus precursor protein (TpP), and D-dimer. C reactive protein (CRP) and cardiac Troponin I (cTnI) have also been determined and compared. Patients in the UA Group were followed for in-hospital adverse events (sudden death, acute myocardial infarction and angina refractory to medical therapy). CRP, D-dimer and cTnI plasma levels were significantly lower in the SEA Group than in the UA Group; the same trend was found for fibrinogen and F1 + 2 plasma levels, although not statistically significant. The TpP was similar in all groups. The control group showed the lowest levels for all indicators. Within the UA Group, 17 patients developed adverse events during hospitalization; F1 + 2, D-dimer, cTnI and CRP plasma levels were higher in these patients than in those with good outcome. Relative risks for adverse events associated with the highest tertile of D-dimer, cTnI, and CRP plasma levels were 8.4 (95% confidence interval, 1.5 48.9), 6.7 (95% confidence interval, 1.1-38.6) and 5.2 (95% confidence interval, 1.1-25.2), respectively. D-Dimer is significantly increased in patients with unstable angina and, in particular, in those who develop an adverse event. PMID- 11943941 TI - The Activated Seven Lupus Anticoagulant (ASLA) assay: a new test for lupus anticoagulants (LAs). Evidence that some LAs are detectable only in extrinsic pathway-based assays. AB - Accurate and timely detection of lupus anticoagulants (LAs) is of diagnostic and prognostic importance due to the association of persistent LAs with thrombotic disease. In the present study, a sensitive and specific assay for LAs has been developed using recombinant activated factor VII to initiate in vitro coagulation. The Activated Seven Lupus Anticoagulant (ASLA) assay uses dilute brain-derived phospholipid in the screening test and a platelet neutralization procedure (PNP) in the confirmatory test. Tests are reported as ratios of patient clotting time to normal control clotting time and percentage correction by PNP assessed for abnormal ratios. Evaluation with 70 known LA-positive plasmas demonstrated higher detection rates than with individual assays from a wide range of commonly employed LA tests. The ASLA assay identified 61 of 70 (87%) known LAs, compared with 65.7% with the most sensitive of the other assays. The various LA assays were used to test 110 plasma samples from patients with thrombotic disease previously negative for LA. These experiments demonstrated that 18 of 110 (16.4%) contained LAs detectable only in extrinsic pathway-based assays, 10 of these by ASLA testing alone. ASLA testing showed high diagnostic precision and has the potential to make a significant contribution to LA detection. PMID- 11943942 TI - Hyperhomocysteinemia increases the risk of venous thrombosis independent of the C677T mutation of the methylenetetrahydrofolate reductase gene in selected Brazilian patients. AB - Fasting total homocysteine (tHcy) and the methylenetetrahydrofolate reductase (MTHFR) C677T mutation were evaluated in 91 patients with venous thromboembolism and without acquired thrombophilia, and in 91 age-matched and sex-matched controls. Hyperhomocysteinemia was detected in 11 patients (12.1%) and in two controls (2.2%), yielding an odds ratio (OR) for venous thrombosis of 6.1 [95% confidence interval (CI), 1.3-28.4]. After excluding 21 patients and four controls with other known genetic risk factors for venous thrombosis, the OR was not substantially changed (7.0; 95% CI, 1.5-33.1). The prevalence of the MTHFR 677TT genotype was not significantly different in patients (9.9%) and in controls (5.5%), with an OR for venous thrombosis of 1.8 (95% CI, 0.6-5.8). Subjects with the MTHFR 677TT genotype showed higher levels of tHcy compared with the 677CC genotype in patients (P = 0.010) and in controls (P = 0.030). In conclusion, we found that fasting hyperhomocysteinemia is a risk factor for venous thrombosis in patients without known acquired thrombophilia and other genetic risk factors for venous thrombosis. Although tHcy levels are significantly higher in those homozygous for the MTHFR C677T mutation, this genotype does not increase the thrombotic risk in our study population. PMID- 11943943 TI - Cyclooxygenase-2 expression in early gastric cancer, intestinal metaplasia and Helicobacter pylori infection. AB - Cyclooxygenase (COX) is the crucial enzyme for synthesis of prostaglandins and occurs in two isoforms COX-1 and COX-2. Whilst COX-1 is constantly expressed in the gastrointestinal tract in large quantities and probably maintains mucosal integrity through constant generation of prostaglandins, COX-2 is induced principally during inflammation. In early gastric cancer and in intestinal metaplasia the expression of COX-2 in patients infected by Helicobacter pylori is increased in intestinal type compared to diffuse type gastric cancer and in intestinal metaplasia. In tumours of mixed type, COX-2 is also increased in the intestinal component compared to the diffuse component. Whilst there has been success of COX-2 inhibition for chemoprevention in colon cancer, a similar role in gastric cancer needs to be carefully assessed in the light of reported adverse effects and whether the precancerous condition, intestinal metaplasia, can truly regress. PMID- 11943944 TI - Why do bacteria reach ascitic fluid? AB - Spontaneous bacterial peritonitis (SBP) is the most common and serious infection that develops in cirrhotic patients. Translocation of bacteria from their intestinal origin, alterations in immune defence mechanisms and deficiencies in the ascitic fluid antimicrobial activity seem to represent the main steps in the pathogenesis of SBP in cirrhosis. Among the factors determining the development of bacterial translocation, intestinal bacterial overgrowth (mainly related to decreased intestinal motility) and changes in the intestinal barrier appear to play an outstanding role. In conclusion, greater understanding of the pathogenesis of SBP will allow better identification of patients at high risk of developing this complication and contribute to the search for new strategies for its prevention. PMID- 11943945 TI - Of liver, whisky and plants: a requiem for colchicine in alcoholic cirrhosis? AB - Colchicine decreases liver fibrosis in experimental and human disease, but a meta analysis recently concluded that colchicine should not be used for liver fibrosis or cirrhosis irrespective of the aetiology. In this issue, Cortez-Pinto et al. confirm such negative conclusions in their series of 55 outpatients with biopsy proven alcoholic cirrhosis followed for a median of 3.5 years. Although well tolerated, colchicine did not affect either the annual incidence rate of complications or liver function tests. Current treatment of alcoholic cirrhosis includes correction of nutritional deficiencies, exogenous administration of antioxidants (notably S-adenosylmethionine and polyenylphosphatidylcholine), and liver transplantation. In the future, preventive/therapeutic strategies will include campaigns to decrease alcohol abuse aimed at subjects genetically prone to develop alcoholic liver injury, prevention of liver fibrosis via inhibition of the Na+/H+ exchange, stimulation of apoptosis of stellate cells, antagonism of cytokines involved in liver injury, degradation of extracellular matrix, and reversal of ethanol-induced inflammatory and fibrotic changes via increased nitric oxide levels. On the grounds that it renders the hepatocyte more vulnerable to necrosis, steatosis has a key role in the pathogenesis of alcoholic and non-alcoholic liver disease. Conditions associated with insulin resistance have been recognized as risk factors for chronic liver disease and hepatocellular carcinoma in the alcoholic. This suggests that, through steatosis, insulin resistance could be a co-factor of alcoholic liver disease. Were such a hypothesis confirmed, it would unify our view of the pathogenesis of alcoholic and non-alcoholic liver disease, with all its inherent therapeutic implications. PMID- 11943946 TI - Cyclooxygenase-2 expression is increased in early intestinal-type gastric cancer and gastric mucosa with intestinal metaplasia. AB - OBJECTIVE: Cyclooxygenase-2 (COX-2) expression is increased in gastric cancer. We examined COX-2 expression in early stage gastric cancer and background mucosa to elucidate the role of COX-2 in gastric carcinogenesis. METHODS: Thirty-three early gastric cancers obtained from 30 patients infected with Helicobacter pylori were studied. Twenty-three patients had an intestinal, four patients had a diffuse, and three patients had both an intestinal and a diffuse type cancer. Expression of COX-2 protein was detected by immunohistochemistry by counting the number of positive staining cells per 100 cells. RESULTS: Mean COX-2 expression was 84.1 (SD 11.4) in 26 intestinal type cancers and was significantly higher than that in seven diffuse type cancers (23.1 +/- 9.7) (P < 0.001). In three patients who had both the intestinal and the diffuse type cancer, COX-2 expression was 92, 90 and 83 in the intestinal type cancer and only 25, 24 and 7 in the corresponding diffuse type cancer. In 18 patients who had intestinal metaplasia (15 had incomplete metaplasia), COX-2 expression was 60.2 (24.2) in the crypts with metaplasia while it was only 16.8 (10.7) in the crypts without metaplasia (P < 0.001). CONCLUSIONS: COX-2 expression may be associated with the carcinogenesis of the intestinal type gastric cancer and, speculatively, inhibition of COX-2 might have preventative effects on the intestinal type gastric cancer. PMID- 11943947 TI - Effects of omeprazole and famotidine on fibroblast growth factor-2 during artificial gastric ulcer healing in humans. AB - BACKGROUND AND AIMS: Endoscopic mucosal resection is a widely accepted technique for the treatment of early gastric cancers, while large ulcers induced by the treatment should be treated promptly. This study aimed to compare the effects of omeprazole and famotidine on ulcer healing and fibroblast growth factor-2 levels in gastric ulcers induced by endoscopic mucosal resection. METHODS: Sixteen patients indicated for endoscopic mucosal resection were enrolled. They were treated by using either omeprazole (n = 8) or famotidine (n = 8) after endoscopic mucosal resection. Endoscopy was performed on days 4, 7 and 28 during each treatment period. Levels of fibroblast growth factor-2 in biopsy specimens were measured by using an enzyme-linked immunosorbent assay at the time of and after endoscopic mucosal resection. Histological variables were also assessed. RESULTS: Ulcer healing rates under endoscopy were not different between the two treatment groups. In both groups, levels of fibroblast growth factor-2 slightly increased on day 4, but the values were not different at any time point. There were no differences in histological variables on days 4 and 7, but fibromuscular hyperplasia was significantly greater in the omeprazole group than in the famotidine group on day 28 (P < 0.05). CONCLUSIONS: Omeprazole and famotidine have an equivalent value for the treatment of ulcers induced by endoscopic mucosal resection. While omeprazole had a more potent effect on fibromuscular hyperplasia than did famotidine, such a difference does not seem to be explained by fibroblast growth factor-2. PMID- 11943948 TI - Ultrastructural characteristics of distal duodenum mucosa in patients with cirrhosis. AB - OBJECTIVES: Morphological abnormalities observed by light microscopy, such as oedema or vasodilatation, have been described in intestinal mucosa of patients with cirrhosis, but no information is available regarding the ultrastructural characteristics of the intestinal epithelial layer. The aim of this observational study was to investigate the ultrastructural characteristics of the intestinal epithelial layer of duodenum mucosa in patients with cirrhosis. METHODS: Six patients with advanced cirrhosis and six control patients without liver disease were included in the study. Biopsies were obtained from the distal duodenum during upper diagnostic endoscopy, and ultrastructural characteristics were studied by means of electron microscopy. RESULTS: A distended interenterocyte space with intestinal epithelial cells closely attached by morphologically intact tight junctions has been observed in cirrhotic patients, together with shorter and wider microvilli than in the control subjects. CONCLUSIONS: The epithelial layers from cirrhotic patients show ultrastructural abnormalities. However, it is not known if the observed alterations are related to the presence of increased intestinal permeability or to bacterial translocation, which are frequently found in these patients. PMID- 11943949 TI - Lack of effect of colchicine in alcoholic cirrhosis: final results of a double blind randomized trial. AB - BACKGROUND: Colchicine, an inhibitor of collagen synthesis, has been suggested as potentially beneficial in cirrhosis. OBJECTIVE: This long-term, randomized, double blind, placebo controlled trial was conducted in order to evaluate the efficacy of colchicine in alcoholic cirrhosis. METHODS: Ambulatory patients with biopsy proven alcoholic cirrhosis, presenting from 1989 to 1997, with no exclusion criteria (e.g. Child-Pugh C, bilirubin > 10 mg/dl and gastrointestinal bleeding in the previous 15 days), were randomized to receive orally, 5 days/week, 1 mg/day of colchicine or placebo. MAIN OUTCOME MEASURES: Results were analysed on an intention to treat basis, for survival, incidence of complications, biochemical liver tests and safety. RESULTS: Twenty-nine patients received colchicine and 26 placebo; characteristics of both groups were similar. The median follow-up was 40.6 (1.4-126.3) months in the colchicine versus 42.4 (5.7-118.2) months in the placebo group (NS). No significant side effects were reported. During follow-up, there were no significant differences in compliance and alcohol abstinence (86% vs 85%). Overall survival was not statistically different (P = 0.38). Cumulative 3-year survival rates were 74.9% in the colchicine versus 91.4% in placebo group (NS). The annual incidence rate of complications was similar with colchicine or placebo: gastrointestinal bleeding, 1.5% vs 1.2%; ascites, 3.7% vs 3.7%; and encephalopathy, 1.0% vs 0.9%. The comparison of changes in biochemical parameters between groups did not show any significant difference. CONCLUSIONS: Although well tolerated, colchicine does not appear to overcome the progression and natural history of long-established alcoholic cirrhosis. PMID- 11943950 TI - Doppler measurements: a surrogate marker of liver fibrosis? AB - OBJECTIVE: The potential diagnostic value of performing Doppler measurements of liver vasculature to assess early stages of liver fibrosis has not been established. Due to the potential clinical impact, this study focused on the correlation between Doppler measurements and histologically proven liver fibrosis. METHODS: Forty-three consecutive patients with chronic viral hepatitis (79% hepatitis C) were enrolled. At the time of liver biopsy, two independent investigators measured maximum and mean blood flow velocity, resistance indices, vessel diameter and blood flow volume in the portal vein, hepatic artery and hepatic veins. All measurements were taken in triplicate. The mean values were correlated to the degree of liver fibrosis using the Ludwig score. RESULTS: Sixty seven per cent of the patients in our study group had no or only mild fibrosis (Ludwig score stage I or II). Thirty-three per cent showed progressive fibrosis or cirrhosis (Ludwig score stage III or IV). There was a large overlap in the Doppler measurements and findings between the various disease stages. No significant changes of Doppler parameters were detected, even between patients with no or mild fibrosis and patients with severe fibrosis (Ludwig score stage III or IV). CONCLUSIONS: Doppler measurements of the portal vein, hepatic artery and hepatic vein(s) are not a valid surrogate marker of liver fibrosis. Nor are Doppler measurements a useful method to estimate the degree of liver fibrosis. PMID- 11943951 TI - Human fetuin/alpha2HS-glycoprotein level as a novel indicator of liver cell function and short-term mortality in patients with liver cirrhosis and liver cancer. AB - OBJECTIVE: Human fetuin/alpha2HS-glycoprotein (AHSG) is synthesized by hepatocytes. We intended to determine whether liver dysfunction or acute phase reaction is dominant in the regulation of its serum concentrations and to see if decreased AHGS levels are associated with short-term mortality. DESIGN: We determined the serum AHSG levels in patients with acute alcoholic, acute A, B, and Epstein-Barr virus hepatitis, alcoholic cirrhosis, and hepatocellular cancer and correlated them to conventional laboratory parameters of inflammation and liver function. Patients were followed for 1 month. METHODS: Serum AHSG was determined by radial immunodiffusion. RESULTS: Compared to controls, significantly lower AHSG levels were found in patients with liver cirrhosis and hepatocellular cancer but not the acute viral hepatitides. Strong positive correlation with serum transferrin, albumin and prothrombin was found. Febrile episodes were not associated with significantly decreased AHSG levels. Concentrations below 300 microg/ml were associated with high mortality rate (52.0%; relative risk, 5.497; 95% confidence interval, 2.472-12.23; P < 0.0001). Of all laboratory parameters studied serum AHSG levels showed the greatest difference between deceased and survived patients with cirrhosis and cancer. Moreover, other acute phase reactants did not differ significantly. The multiple logistic regression analysis indicated that the decrease of serum AHSG is independent of all other variables that were found decreased in deceased patients. CONCLUSIONS: Decreased serum AHSG concentration is due rather to hepatocellular dysfunction than the acute phase reaction and is an outstanding predictor of short-term mortality in patients with liver cirrhosis and liver cancer. PMID- 11943952 TI - Gastric emptying of solids in cirrhotic and peritoneal dialysis patients: influence of peritoneal volume load. AB - INTRODUCTION: A delay in gastric emptying rate has been reported in peritoneal dialysis patients, often normalizing after evacuation of the dialysate. To evaluate the effect of the intraperitoneal volume, we compared this finding with a cirrhotic model in which gastric emptying was studied before and after a large volume paracentesis. METHODS AND DESIGN: We used the 13C-octanoic acid breath test to measure gastric half-emptying time (T1/2) for solids in patients with alcoholic cirrhosis, non-diabetic peritoneal dialysis patients, and a control population (asymptomatic volunteers). Cirrhotic patients underwent the test on two consecutive mornings before and after an evacuating paracentesis. Peritoneal dialysis patients were studied twice on consecutive days: once with the dialysate present intra-abdominally ("full"), and once with an emptied abdomen ("empty"). Biochemical analysis was carried out on blood samples before the first test. All cirrhotics underwent a 13C-aminopyrine breath test to assess residual liver function. RESULTS: Gastric emptying in cirrhotics showed no difference before or after paracentesis (median T1/2 108.0 min v. 117.9 min), but it was delayed significantly versus normal in both tests. There was no correlation with biochemical parameters, Child-Pugh score, or 13C-aminopyrine breath test results. Gastric half-emptying times of "full" peritoneal dialysis patients (median T1/2 103.1 min) were significantly higher than those of "empty" peritoneal dialysis patients (median T1/2 68.9 min) and asymptomatic volunteers (median T1/2 60.1 min). "Empty" peritoneal dialysis patients showed no gastroparesis. CONCLUSION: In alcoholic cirrhotic patients with ascites, gastric emptying of solids is delayed, independently of the volume of ascites. In peritoneal dialysis patients, gastric emptying was delayed when "full" and normalized after drainage of the dialysate. PMID- 11943953 TI - Effect of cholecystectomy for gallstones on the surface of the papilla of Vater and the diameter of the common bile duct. AB - BACKGROUND AND STUDY AIMS: To date, little is known about changes in the size of the papilla of Vater in patients with gallstones. Most of the research concerning these patients has investigated changes in the diameter of the common bile duct and pressure in the sphincter of Oddi region. The latest research has not confirmed the prevalent opinion that the common bile duct dilates after cholecystectomy; moreover, knowledge about changes in the form and size of the papilla of Vater remains poor. The aim of our research was to measure the surface of the papilla of Vater and the diameter of the periampullary pancreatic part of the common bile duct before surgery, then 3 months and 6 months after surgery, using endoscopic ultrasound. PATIENTS AND METHODS: In patients with symptomatic gallstones but with no evidence of cholestasis or stones in the common bile duct, the surface of the papilla of Vater and the diameter of the periampullary pancreatic part of the common bile duct were measured before surgery by endoscopic ultrasound. In the postoperative period, measurements were repeated in patients with an early occurrence of nausea, distension, bloating and pain in the upper abdomen and in patients without any symptoms in the same time period. The surface of the normal papilla of Vater and the diameter of the periampullary pancreatic part of the common bile duct were determined in healthy subjects. RESULTS: In all the patients included in our study, the surface of the papilla of Vater was enlarged before surgery and, after 3 months, was even larger. However, after 6 months, it was practically the same as before surgery. The diameter of the periampullary pancreatic part of the common bile duct before surgery and 6 months after surgery was within normal limits. However, a statistically significant and transient increase in the diameter was noticed in the third month after cholecystectomy. The results were almost the same for symptomatic and asymptomatic patients. CONCLUSION: In patients with symptomatic gallstones, the surface of the papilla of Vater is increased while the diameter of the periampullary pancreatic part of the common bile duct is normal. Cholecystectomy is an intervention that causes an additional transient increase in the surface of the papilla of Vater and a transient increase in the diameter of the periampullary pancreatic part of the common bile duct. The surface of the papilla of Vater does not reach normal values but the diameter of the periampullary pancreatic part of the common bile duct is normalized 6 months after surgery. There is no relation between clinical disorders and changes in the morphology of the papilla of Vater and the periampullary pancreatic part of the common bile duct after cholecystectomy. PMID- 11943954 TI - Circulating soluble vascular adhesion protein 1 in patients with inflammatory bowel disease. AB - OBJECTIVE: A dysregulated local immune defence with a constant influx of leucocytes provides a basis for continuous intestinal inflammation in ulcerative colitis and Crohn's disease. Since vascular adhesion protein 1 (VAP-1) is one of the adhesion molecules that mediates lymphocyte binding to endothelium, we investigated the levels of soluble VAP-1 (sVAP-1) in the sera of inflammatory bowel disease (IBD) patients compared with healthy controls. METHODS: sVAP-1 serum levels were measured in 161 IBD patients (90 ulcerative colitis, 71 Crohn's disease) and 93 controls using a commercially available enzyme-linked immunosorbent assay (ELISA). sVAP-1 levels were correlated with disease activity and localization. In 42 patients, sVAP-1 levels were measured in both the active and inactive phases of the disease. RESULTS: sVAP-1 serum levels were detected in all control and IBD subjects. Mean sVAP-1 levels were 365.5 +/- 153.5 ng/ml in ulcerative colitis patients, 336.4 +/- 172.8 ng/ml in Crohn's disease patients, and 344.7 +/- 150.4 ng/ml in healthy controls. The differences between the groups were not significant. No association between disease activity or disease localization and sVAP-1 was found. CONCLUSIONS: sVAP-1 serum concentrations are not significantly different in IBD and healthy control subjects. sVAP-1 serum levels are of no value in the assessment of disease activity or severity of inflammation in patients with IBD. PMID- 11943955 TI - A new, highly sensitive assay for C-reactive protein can aid the differentiation of inflammatory bowel disorders from constipation- and diarrhoea-predominant functional bowel disorders. AB - BACKGROUND: Patients presenting to gastroenterology clinics with symptoms suggestive of lower-bowel disorders often require extensive investigation to differentiate functional from organic disease. C-reactive protein (CRP) is a sensitive marker of systemic inflammation. Levels of CRP are frequently raised in cases of inflammatory bowel disease (IBD). However, using conventional assays, not all cases of IBD have a detectable level. OBJECTIVE: To determine whether a new highly sensitive CRP enzyme-linked immunosorbent assay (ELISA) can aid the differentiation between IBD and functional bowel disorders (FBDs) in gastroenterology outpatients presenting with lower-bowel symptoms. METHODS: Serum was taken from 224 subjects attending a gastroenterology outpatient clinic. Of these, 203 were new patients and 21 were follow-up patients with quiescent colitis. The serum was analysed using a sensitive in-house ELISA. All new patients had a rigid sigmoidoscopy and rectal biopsy. Patients were investigated as deemed appropriate by the attending physician. Notes were reviewed after at least 6 months to determine the final diagnosis. RESULTS: A cut-off value of 2.3 mg/l had a sensitivity of 100% and a specificity of 67% in differentiating FBD from new cases of IBD. The geometric mean CRP was 0.383 mg/l in the constipation predominant FBD group, 1.435 mg/l in diarrhoea-predominant FBD, 1.455 mg/l in quiescent IBD, 8.892 mg/l in newly presenting cases of ulcerative colitis, and 13.123 mg/l in newly presenting cases of Crohn's disease. CONCLUSION: A new, highly sensitive assay for CRP may help to distinguish FBD from IBD. PMID- 11943956 TI - Persistent activation of coagulation and fibrinolysis after treatment of active ulcerative colitis. AB - OBJECTIVE: Haemostatic changes may be involved in the pathogenesis and progression of ulcerative colitis. We studied longitudinally inflammatory and haemostatic parameters in patients treated for severe ulcerative colitis. DESIGN AND SETTING: We carried out a descriptive study of longitudinal blood measurements in patients with severe ulcerative colitis from one large regional hospital. METHODS: Nineteen patients with severe ulcerative colitis were assessed by an endoscopic score and a patient score at baseline. Patients were assessed by patient scores during treatment at scheduled intervals. At each visit, inflammatory and haemostatic parameters were determined. RESULTS: At baseline, the erythrocyte sedimentation rate, C-reactive protein, leucocyte and granulocyte count, thrombin-antithrombin complexes, prothrombin fragment 1+2, fibrinogen and degradation products of fibrinogen and fibrin were increased in patients when compared with controls, whereas albumin concentration and factor XIII activity were significantly lower. Antithrombin activity was normal. During treatment, the median patient score diminished significantly from 12 to 4.5 points after 2 weeks, decreased further to 4 points after 4 weeks and remained below 4 points throughout the remaining study period. Inflammation parameters returned to within the reference range in two patients after 4 weeks, whereas the coagulation markers prothrombin fragment 1+2 and thrombin-antithrombin complexes returned to normal values after 8 weeks and 24 weeks, respectively. In contrast with markers of inflammation, slightly increased concentrations of the degradation products of both fibrinogen and fibrin were found for almost 1 year, which indicated low grade activation of coagulation and fibrinolysis. CONCLUSIONS: These results are compatible with a condition of persistent hypercoagulation in patients with ulcerative colitis who are in clinical remission. Persistent hypercoagulation may contribute to the clinical course of ulcerative colitis. PMID- 11943957 TI - Management of Lithuanian children's acute diarrhoea with Gastrolit solution and dioctahedral smectite. AB - OBJECTIVE: Acute gastroenteritis represents a major cause of morbidity and mortality worldwide among children, and rehydration treatment has been one of the cornerstones in the management strategy. The natural clay dioctahedral smectite (Smecta) increases intestinal barrier function and is effective against infectious diarrhoea in children. The purpose of this work was to compare the efficacy and tolerance of Lithuanian children's diarrhoea treatment with dioctahedral smectite combined with hypotonic oral rehydration solution (ORS)- Gastrolit--versus Gastrolit alone to establish the influence of Smecta on serum electrolyte balance in young children with diarrhoea and mild or moderate dehydration. METHODS: Smecta combined with ORS (study group) and ORS alone (control group) were evaluated in a multicentre, open, randomized trial in 54 children aged 6-48 months hospitalized for acute diarrhoea (mostly rotavirus aetiology) and signs of mild and moderate dehydration. The main outcomes examined were duration of diarrhoea, fever, number of vomiting episodes, and serum electrolyte balance before and after treatment. RESULTS: The mean duration of diarrhoea was significantly shorter in the study group (42.3 +/- 24.7 h) than in the control group (61.8 +/- 33.9 h). No side effects of Smecta were observed. The changes of sodium, potassium, chloride and calcium concentrations after treatment were minimal and in the normal range. CONCLUSIONS: Smecta significantly reduced the duration of diarrhoea, was safe and well tolerated, and had no impact on the adsorption of electrolytes. Smecta could be used together with ORS in children suffering from acute gastroenteritis (without uncontrollable vomiting) with mild and moderate dehydration. PMID- 11943958 TI - Low serum vitamin B12 is common in coeliac disease and is not due to autoimmune gastritis. AB - OBJECTIVE: Although coeliac disease is a disorder of the proximal small bowel, associated vitamin B12 deficiency has been reported. This study aimed to assess the prevalence of B12 deficiency in a large series of coeliac patients, and to exclude the possibility that it is due to associated autoimmune gastritis. DESIGN: Prospective routine measurement of serum B12 in coeliac patients, with investigations for pernicious anaemia/autoimmune gastritis in B12-deficient patients. SETTING: Gastroenterology department of a large district general hospital. INTERVENTIONS: If they were not taking vitamin B12 supplements already, patients had serum B12 measured before starting dietary gluten exclusion. Those with low levels also had gastric biopsies taken and plasma gastrin and serum gastric parietal cell and intrinsic factor antibodies measured. MAIN OUTCOME MEASURES: Prevalence of low serum B12, and presence or absence of indicators of pernicious anaemia/autoimmune gastritis in patients with low serum B12. RESULTS: Of 159 patients, 13 had low serum B12 at diagnosis. A further six had been receiving B12 replacement therapy for 3-37 years before diagnosis, giving an overall prevalence of 12% (19 patients). Only 2/19 patients had gastric corpus atrophy, one with intrinsic factor antibodies and the other with hypergastrinaemia. There was no relationship between low B12 and clinical characteristics. CONCLUSIONS: Low B12 is common in coeliac disease without concurrent pernicious anaemia, and may be a presenting manifestation. B12 status should be known before folic acid replacement is started. PMID- 11943959 TI - Establishing patient preferences for gastroenterology clinic reorganization using conjoint analysis. AB - OBJECTIVE: The Department of Health states that patients with suspected cancer should be seen within 2 weeks, and the Patients' Charter suggests that patients should not wait for more than 30 min in outpatients. Decisions such as these are often made with little assessment of patient preferences. We have elicited patient preferences for the optimal use of time in the outpatient clinic. DESIGN: Questionnaire survey eliciting preference between different clinic scenarios evaluated using discrete choice conjoint analysis. SETTING AND PARTICIPANTS: Patients attending a teaching hospital gastroenterology outpatient clinic. MAIN OUTCOME MEASURES: The relative importance of time spent on the waiting list, time waiting in the clinic, time spent with the specialist, and time waiting for investigation was assessed using a logit model. RESULTS: Patients placed a similar value on waiting for investigation and time spent on the waiting list. A clinic that had a 2-month waiting list but offered immediate investigations would therefore be more popular than a clinic that had a 2-week waiting list but whose investigations were deferred for 3 months. Patients would be prepared to spend an extra 30 min in the waiting room if they spent 1 month less on the waiting list or waiting for investigation. Time spent with a specialist is valued, and patients would be prepared to spend an extra 3 min waiting in the clinic for every extra minute spent with the doctor. CONCLUSIONS: The present Department of Health recommendations and the Patients' Charter are too simplistic and do not take into account patient preferences. PMID- 11943960 TI - High-flow-rate haemodiafiltration as a brain-support therapy proceeding to liver transplantation for hyperacute fulminant hepatic failure. AB - In fulminant hepatic failure (FHF), rapidly progressive cerebral oedema remains the main fatal complication and an obstacle in liver transplantation. A 29-year old Japanese woman presented with sudden-onset hepatic encephalopathy and jaundice. Hepatic encephalopathy deteriorated within 2 days of the onset of jaundice. She manifested extensory sustained clonus and was responsive only to pain. Diffuse cerebral oedema was noted on brain computerized tomography (CT) scan. Urgent living-donor liver transplantation (LDLT) at the time of admission was abandoned because of deterioration of neurological status and radiologically evident diffuse cerebral oedema. Instead, a high-flow-rate (7.2-9.0 l/h) haemodiafiltration with a high-performance membrane was commenced, combined with plasma exchange. This treatment regimen resulted in a gradual improvement of hepatic encephalopathy and complete disappearance of cerebral oedema within 7 days. Liver regeneration did not occur during this period, as evident by CT scan volumetry and serological tests. LDLT was subsequently performed using the right liver lobe of the patient's brother. Our case suggests that high-flow-rate haemodiafiltration with a high-performance membrane, combined with plasma exchange, could potentially be brain-support therapy for patients with FHF, and may contribute, when combined with liver transplantation, to the improvement of prognosis in hyperacute FHF. PMID- 11943961 TI - Paracetamol poisoning unmasking factor VII deficiency. AB - We report on an 18-year-old woman who took an impulsive overdose of paracetamol. The admission international normalized ratio (INR) was prolonged which resulted in her being transferred to a specialist hepatology unit. Her clinical condition and laboratory parameters did not correlate; this prompted investigation into her coagulation profile, which revealed a factor VII deficiency and explained the observed abnormalities of an elevated INR, normal partial thromboplastin time and normal liver function. There are no other reports of factor VII deficiency being identified as a result of paracetamol overdose. PMID- 11943962 TI - Gastrointestinal stromal tumour of the oesophagus: significance of immunohistochemical and genetic analyses of the c-kit gene. AB - Oesophageal gastrointestinal stromal tumours (GISTs) are rare in comparison to those of the stomach and intestines. Recently, it has been clarified that mutations of the c-kit gene resulting in gain of function might be associated with histogenesis of this type of tumour arising in the stomach and intestines. We describe an oesophageal GIST on immunohistochemical and genetic analyses of the gene. A 71-year-old man had an intramural tumour of the middle third of the oesophagus. Tumour cells were composed predominantly of spindle-shaped and partially epithelioid cells. They were diffusely positive for CD117. Six base deletion resulting in in-frame mutation of the c-kit gene was confirmed at codon 556-558 (cag tgg aag to cag) of exon 11. Patients with mutations of the c-kit gene revealed worse prognoses in GISTs arising from other locations. A long-term follow-up observation is needed for the case. PMID- 11943963 TI - Two cases of adult T-cell leukaemia/lymphoma with oesophageal involvement. AB - We describe two cases of adult T-cell leukaemia/lymphoma (ATLL) with oesophageal involvement. The first case, a 51-year-old Japanese woman with an acute subtype of ATLL, had an irregular ulcerative lesion in the distal oesophagus. The second case, a 76-year-old Japanese man with a lymphoma subtype of ATLL, had a polypoid lesion in the middle portion of the oesophagus. Both cases had gastric involvement. Biopsies from these lesions revealed mucosal invasion of ATLL cells in each case. Combination chemotherapy was ineffective in both cases. Prospective and careful examination of additional cases may eventually provide specific advice for treatment of this unusual condition. PMID- 11943965 TI - A case of pancreaticoduodenal artery aneurysm causing pancreatic pseudotumour and duodenal obstruction. AB - This case report describes a ruptured pancreaticoduodenal artery aneurysm (PDAA) causing pancreatic pseudotumour and duodenal obstruction. A 59-year-old man was referred to our hospital with a chief complaint of frequent vomiting without abdominal pain. Because a mass lesion 10 cm in diameter was palpated in the right para-umbilical region and found in the head of the pancreas on computerized tomography (CT) and ultrasonography, malignant tumour of the pancreas or tumour forming pancreatitis was strongly suspected, and further examination was performed.Magnetic resonance imaging (MRI) results suggested subacute haematoma inside the mass. On angiography, an aneurysm 8 mm in diameter was found in the posterior superior pancreaticoduodenal artery (PSPD). Since an ultrasound-guided percutaneous needle biopsy from the solid part of the mass indicated no malignancy, the lesion was considered an inflammatory pseudotumour in the head of pancreas due to ruptured aneurysm. Bypass surgery was planned, but the tumour shrank significantly with conservative treatment. Obstruction disappeared completely without surgery 4 weeks after the first symptom. PMID- 11943964 TI - Selective colonization by Helicobacter pylori of the deep gastric glands and intracellular canaliculi of parietal cells in the setting of chronic proton pump inhibitor use. AB - Colonization by Helicobacter pylori of the acid-secreting tubules of gastric glands and the canaliculi of parietal cells has only rarely been reported. The presence of these organisms in such "deep" locations has only been reported in association with the more typical superficial colonization of the mucous gel layer overlying gastric epithelial cells. We report two cases of deep H. pylori infection without the presence of superficial organisms. Both patients had been using proton pump inhibitors for many years. We review the literature regarding the distribution of H. pylori within the stomach and the effect of proton pump inhibitor use on H. pylori distribution. PMID- 11943966 TI - Idiopathic hepatocellular adenoma with intratumoral haemorrhage. PMID- 11943967 TI - Potentially harmful sleep habits of 3-year-old children in Japan. AB - To examine the sleep habits of 3-year-old children, we questioned guardians during a routine health examination for 3-year-old children at a public health center. According to the 1105 questionnaires analyzed, the proportion of children who fell asleep at 10 p.m. or later was 49.6%. The nocturnal sleep onset time was significantly correlated with the wake-up time in the morning and was significantly negatively correlated with the nocturnal sleep duration. The average daily total sleep duration (nocturnal sleep duration + nap duration) of regular nap-takers showed a significant negative correlation with the nocturnal sleep onset time. The average values for height, weight, and body mass index (BMI) were not correlated with the nocturnal sleep onset time. Children who went to sleep later got less sleep than those who went to sleep earlier. Because sleep debt has a harmful impact on older children and adults, late sleep onset may have adverse health consequences in young children. PMID- 11943968 TI - Teaching firearm safety to children: failure of a program. AB - The present study investigated the effectiveness of a skills-based firearm safety program on reducing children's play with firearms. In a randomized control study, 34 children aged 4 to 7 years participated in a week-long firearm safety program; the Control Group was composed of 36 children. After the program, pairs of children were observed playing in a structured setting in which they had access to a semiautomatic pistol. A total of 53% of the pairs played with the gun, and there was no difference in gun-play behavior between those children who did and did not receive the intervention. Interview data revealed significant discrepancies in parent and child reports of parental gun ownership and inaccurate parental predictions of their children's interest in guns. The results of the current study cast doubt on the potential effectiveness of skills-based gun safety programs for children. PMID- 11943969 TI - Problem-solving skills training for mothers of children with newly diagnosed cancer: a randomized trial. AB - Mothers of children with serious illnesses have lower levels of well-being than mothers in the general population. Problem-solving therapy (PST), a cognitive behavioral intervention, has been shown to be effective in treating negative affectivity (depression, anxiety) and other manifestations of reduced well-being. This report describes a problem-solving skills training (PSST) intervention, based on problem-solving therapy, for mothers of newly diagnosed pediatric cancer patients. Ninety-two mothers were randomly assigned to receive PSST or to receive standard psychosocial care (Control Group). After the 8-week intervention, mothers in the PSST Group had significantly enhanced problem-solving skills and significantly decreased negative affectivity compared with controls. Analysis revealed that changes in self-reports of problem-solving behaviors accounted for 40% of the difference in mood scores between the two groups. Interestingly, PSST had the greatest impact on improving constructive problem solving, whereas improvement in mood was most influenced by decreases in dysfunctional problem solving. The implications of these findings for refinement of the PSST intervention and for extension to other groups of children with serious illnesses are discussed. PMID- 11943971 TI - Racing cain. PMID- 11943970 TI - Ongoing maternal drug use, parenting attitudes, and a home intervention: effects on mother-child interaction at 18 months. AB - This prospective study examined the effects of ongoing maternal drug use, parenting attitudes, and a home-based intervention on mother-child interaction among drug-using women and their children. At 2 weeks postpartum, mothers and infants were randomly assigned to either an Intervention (n = 67) or Control (n = 64) Group. Intervention families received weekly visits until 6 months postpartum and biweekly visits from 6 to 18 months by trained lay visitors. The home intervention was designed to increase maternal empowerment and promote child development. Control families received brief monthly tracking visits. Mother child interaction was evaluated at 18 months through observation of play. Mothers who continued to use cocaine and/or heroin had lower competence scores (p <.05); poor parenting attitude was also associated with lower competence scores during mother-child interaction (p <.05). Although the intervention had no measured effect, ongoing maternal drug use and poor parenting attitudes were associated with less optimal maternal behavior during mother-child interaction. PMID- 11943972 TI - "Not listening, stubborn, and problems with sleeping"--interpreting a parent's concerns at a 3 year old's health supervision visit. PMID- 11943973 TI - Associations between physiological reactivity and children's behavior: advantages of a multisystem approach. AB - The past decade has seen a notable increase in interest in and research concerning the physiological correlates of behavior problems in childhood. The present article reviews what this growing body of research has revealed. The main tenet is that disruptions in both sympathetic and adrenocortical regulation appear to be common among children with internalizing and externalizing behavior problems. The associations between such neuroendocrine alterations and behavior are discussed and their implications for the fields of stress physiology, neuroendocrinology, and developmental psychopathology are outlined. It is proposed that substantial advances can be made by investigating patterns of physiological responses among multiple, concurrent systems rather than individual response systems. PMID- 11943974 TI - Measuring the results of an isolated therapeutic intervention does not adequately measure the effectiveness of all motor intervention. PMID- 11943979 TI - Prevention in pediatric orthopaedics. PMID- 11943980 TI - Early weaning might reduce the psychological strain of Boston bracing: a study of 136 patients with adolescent idiopathic scoliosis at 3.5 years after termination of brace treatment. AB - From 1983 to 1990 a total of 136 patients with adolescent idiopathic scoliosis were treated with the Boston brace. With the aim of examining the social and psychological impact of the brace treatment all patients received a questionnaire at an average of 3.5 years after termination of the treatment. Ninety-three percent responded to the questionnaire. The overall picture was that the patient's daily activities and social contacts, not least with the opposite sex, were affected both during the bracing period and at follow-up. It is important that the indications for the treatment are assured, and weaning should be initiated at the latest 3 years after menarche. PMID- 11943981 TI - Perthes-like changes in geleophysic dysplasia. AB - Geleophysic dysplasia is a rare skeletal dysplasia characterised by 'happy natured' facies, short stature with short limbs, brachydactyly, and joint contractures. This report describes a case of a patient with unilateral Legg Calve-Perthes-like changes associated with dysplastic proximal capital femoral epiphysis, typical to geleophysic dysplasia. PMID- 11943982 TI - Bioabsorbable rods in Salter's osteotomy. AB - Salter's innominate osteotomy is a commonly used procedure in the treatment of developmental dysplasia of the hip. The graft is usually fixed with two K-wires that are removed surgically 6 weeks later. The use of bioabsorbable rods to fix the graft saves patients further surgery for removal of wires and ensures a better scar. This paper reports for the first time the successful use of bioabsorbable self-reinforced polyglycolic acid rods for fixing the graft of Salter's innominate osteotomy in the treatment of developmental dysplasia of the hip, and describes the suggested operative technique for their use. PMID- 11943983 TI - Arthrographic findings in Legg-Calve-Perthes disease. AB - Plain radiographs and arthrograms on 46 patients with Legg-Calve-Perthes were evaluated. Measurements of containment including epiphyseal extrusion, caput index, femoral subluxation, percentage acetabular coverage, were determined. Only with arthrography are measurements of acetabular coverage, subluxation and epiphyseal extrusion consistent and reliable. The concept of eccentricity was developed to assist in the understanding of femoral containment. PMID- 11943984 TI - Adductor myositis as a cause of childhood hip pain. AB - Two cases of bacterial adductor myositis that presented as painful hips in boys aged 4 and 9 years are reported. Clinically, there was severe pain and a high pyrexia, a raised C-reactive protein and positive blood cultures but a negative hip ultrasound. Urgent magnetic resonance imaging demonstrated changes throughout the adductor muscles in keeping with bacterial myositis. Both boys settled with intravenous antibiotic therapy. We propose that magnetic resonance imaging is a valuable tool in the assessment of infection around the hip and should be indicated when other investigations have excluded a septic arthritis but the child remains unwell. PMID- 11943985 TI - Correction of foot deformities by the Ilizarov method without corrective osteotomies or soft tissue release. AB - Twenty-three foot deformities in 22 patients had been treated without any real surgical incision, 'the bloodless technique', after the application of the Ilizarov external fixator. The ages of the patients ranged from 2 years and 6 months to 49 years. After an average follow-up of 18.3 months (range 8-40 months), all patients had a plantigrade foot. There was no need for soft tissue release or osteotomy. Pin tract infection occurred mostly in the metatarsal wire. However it did not affect final results. The bloodless technique is an effective way of treating foot deformities in children and adults. PMID- 11943986 TI - Seasonal distribution of idiopathic congenital talipes equinovarus in Scotland. AB - Idiopathic congenital talipes equinovarus has a reported prevalence of between 0.64 and 2.5 per thousand live births in the Scottish population. A retrospective study of neonates (n=162), who presented with idiopathic congenital talipes equinovarus in Scotland over a 3 year period (1994-1996) revealed a seasonal increased incidence of the condition in neonates born in March and April. The probable aetiological factors for congenital talipes equinovarus are discussed. PMID- 11943987 TI - Assessment of the position of the navicular by three-dimensional magnetic resonance imaging in infant foot deformities. AB - As the navicular is unossified in infants with foot deformities, until 5 years old (boys) and 3 years old (girls), it is difficult to give the exact position of hindfoot bones. Magnetic resonance imaging associated with three-dimensional reconstruction allows the visualization of the cartilaginous volume of the bones and the measurement of the relationships of the navicular with the talus. In normal feet, we noted that in the coronal plane the navicular was laterally situated (75%) relative to the talus. In club feet (17 patients), the navicular and the cuboid had a large medial displacement (77%) with a slight pronation movement in the sagittal plane. In suspected skewfoot (seven feet) we noted three cases with a lateral subluxation of the navicular. This objective technique of measurement may be of value for diagnosis and treatment of foot deformities. PMID- 11943989 TI - Stabilization of a short juxta-articular bone segment with a circular external fixator. AB - The objective of the current study was to evaluate the stabilization of a simulated juxta-articular bone segment with a circular external fixator, and to determine which method of fixation improved bending stabilization while preserving the axial dynamization of a three-wire configuration. Frames were divided into three groups: wire, half-pin and hybrid and tested in axial compression, torsion, anteroposterior bending and mediolateral bending. Hybrid frames using 4 mm half-pins improved the anteroposterior stabilization of the short bone segment while maintaining axial characteristics similar to a three wire frame. Increasing the bending stabilization will improve bone segment alignment while permitting axial micromotion beneficial to bone healing. PMID- 11943988 TI - Percutaneous fixation in clubfoot surgery: a radiographic and gait study. AB - Twenty patients with Type II clubfeet were evaluated an average of 3.1 years after complete surgical soft tissue release. A pinned group had fixation of the talonavicular and calcaneocuboid joints and a non-pinned group had no fixation. The control group consisted of 10 children without foot ailments. Radiographic measurements of talocalcaneal and talus-first metatarsal angles and frequencies of dorsal and medial navicular subluxation were not different for the two treatment groups. Foot progression angle was not significantly different for the two treatment groups, but was significantly different between all clubfoot patients (3.6 degrees of in-toeing) and the control group (5.0 degrees of out toeing). Tourniquet time was significantly greater for the pinned than for the non-pinned group, but the duration of surgery was similar. Similar radiographic and gait measurement findings suggest that clubfoot surgery with and without percutaneous fixation have comparable outcomes. PMID- 11943990 TI - Skeletal traction and delayed percutaneous fixation of complicated supracondylar humerus fractures due to delayed or unsuccessful reductions and extensive swelling in children. AB - The functional and radiological results of 13 supracondylar humerus fractures of 13 children were evaluated to determine the efficiency of delayed percutaneous fixation of the fractures after a period of skeletal overhead traction. All the fractures were complicated by delayed reduction, extensive swelling or unsuccessful reduction manipulations. As the unfavourable preexisting factors subsided under skeletal traction, anatomical reduction and fixation of fractures by percutaneous pinning were performed under general anesthesia. The average follow-up period was 21 months. Bauman and lateral capitellohumeral angles were measured and statistically analyzed. Functional and cosmetic results were evaluated by physical examination of the elbows. There were 11 excellent and two fair functional results while all cosmetic results were excellent or good. As a result, skeletal traction and delayed percutaneous pinning is accepted as an alternative method of treatment for complicated supracondylar humerus fractures in children. PMID- 11943991 TI - Atypical growth plate closure: a possible chronic Salter and Harris Type V injury. AB - Salter-Harris Type V epiphyseal injury is relatively uncommon. The authors present a case involving the proximal tibia diagnosed both by radiologic and histologic findings. The findings suggest that Salter-Harris Type V injury can be a possible outcome when the disappearance of the physeal plate is reported in a patient who has great physical activity in childhood, even if the patient has suffered from no major injury. PMID- 11943992 TI - Effect of derotation osteotomy of the femur on hip and pelvis rotations in hemiplegic and diplegic children. AB - The purpose of this study was to evaluate hip and pelvis rotations in groups of hemiplegic and diplegic children before and after surgical correction of fixed internal rotation deformity of the hip. Twenty-two children with cerebral palsy (eight diplegia, 14 hemiplegia) having fixed internal rotation deformity at the hip were treated by multilevel surgery which included derotation osteotomy of the femur. Evaluation was done before and at a mean of 3.1 years after surgery using three-dimensional computerized gait analysis. Preoperatively, the patients in the hemiplegia group had a significantly greater compensatory external rotation of the pelvis than those in the diplegic group. Post-operatively there were no significant differences between the two groups. In the hemiplegia group the external rotation of the pelvis was corrected after correction of hip rotation by derotation osteotomy. Patients in the diplegia group showed significant improvements in the hip rotation with no significant change in the pelvis rotation after multilevel surgery. PMID- 11943993 TI - Tibia valga due to focal fibrocartilaginous dysplasia: case report. AB - Focal fibrocartilaginous dysplasia is a rare benign entity, previously described as a factor responsible for causing tibia vara at the walking age. We report here the case of a child with tibia valga due to focal fibrocartilaginous dysplasia of the lateral part of the proximal tibia. To our knowledge, this is the first observation of a lesion located in the lateral proximal tibia. Clinical, radiographic and magnetic resonance imaging surveillance was performed. Spontaneous resolution of the lesion with correction of the angular deformity did occur. PMID- 11943994 TI - Concomitant focal fibrocartilaginous dysplasia of the tibia and eosinophilic granuloma of the jaw in a child. AB - This 2-year-old child presented with concomitant eosinophilic granuloma of the lower jaw and focal fibrocartilaginous dysplasia of the right tibia. Her eosinophilic granuloma was diagnosed on the basis of the clinical picture, imaging studies and the characteristic histologic appearance. Focal fibrocartilaginous dysplasia was revealed incidentally during the eosinophilic granuloma staging process. After chemotherapy, all signs of eosinophilic granuloma subsided, but focal fibrocartilaginous dysplasia remained without signs of clinical or radiographic progression. The importance of differentiating these two conditions is stressed in order to avoid ineffective and inappropriate treatment of focal fibrocartilaginous dysplasia. PMID- 11943995 TI - Distal fibular reconstruction with pedicled vascularized fibular head graft: a case report. AB - We present an original technique for reconstruction of the distal fibula after tumor resection. The case report was of a 13-year old boy suffering from an osteogenic sarcoma of the distal fibula. Pedicled vascularized epiphyseal transfer using the ipsilateral proximal fibula was performed. Results were assessed at 2 years and 6 months and provided good stability and normal functional outcome of the ankle. PMID- 11943996 TI - Treatment of developmental dysplasia of the hip with the Pavlik Harness: factors for predicting unsuccessful reduction. PMID- 11943998 TI - The business of infection control and epidemiology. PMID- 11943999 TI - The business of health care epidemiology: creating a vision for service excellence. AB - To achieve service excellence in infection control in an era of fiscal constraint requires infection control specialists to demonstrate the value of their department in business terms to their health care administrators. Models for achieving service excellence in infection control based on cost-effective interventions found effective in our organization are described, including development and maintenance of effective teams, guidelines for structuring and implementing interventions, and establishment of business standards for excellence in health care. PMID- 11944000 TI - Making the business case for infection control: pitfalls and opportunities. PMID- 11944001 TI - Organizational and environmental factors that affect worker health and safety and patient outcomes. AB - This article reviews organizational factors that influence the satisfaction, health, safety, and well-being of health care workers and ultimately, the satisfaction, safety, and quality of care for patients. The impact of the work environment on working conditions and the effects on health care workers and patients are also addressed. Studies focusing on worker health and safety concerns affected by the organization and the physical work environment provide evidence of direct positive and/or adverse effects on performance and suggest indirect effects on the quality of patient care. The strongest links between worker and patient outcomes are demonstrated in literature on nosocomial transmission of infections. Transmission of infections from worker to patient and from patient to patient via health care worker has been well documented in clinical studies. Literature on outbreaks of infectious diseases in health care settings has linked the physical environment with adverse patient and worker outcomes. An increasing number of studies are looking at the relationship between improvement in organizational factors and measurable and positive change in patient outcomes. Characteristics of selected magnet hospitals are reviewed as one model for improving patient and worker outcomes. PMID- 11944002 TI - From vision to reality: strategic agility in complex times. AB - Health care is experiencing turbulent times. Change has become the constant. Complexity and sometimes chaos are common characteristics. Within this context, infection control professionals strive to maintain their practices, achieve excellence, and plan for the future. As demands shift and expectations increase, professionals in infection surveillance, prevention, and control (ISPC) programs must develop strategic agility. This article describes the rationale for strategic thinking and action set within a framework of 6 thought-provoking questions. It also describes a number of techniques to use for thinking strategically, such as designing visions, becoming entrepreneurial, and engaging in creative and futuristic exercises to evaluate possibilities for program direction. These techniques can guide individual professionals or ISPC programs in strategic decision-making that will increase the ability to survive and succeed in the future. PMID- 11944003 TI - From expert data collectors to interventionists: changing the focus for infection control professionals. AB - The current economic and political environments challenge health care organizations in the United States to provide affordable, accessible, and comprehensive health services. However, changes in reimbursement to health care providers can affect their ability to offer access to cutting-edge services while reducing costs. Consequently, organizations are restructuring, re-engineering, right-sizing, downsizing, and redesigning, all in an effort to save money while also hoping to maintain a reputation for quality and customer service. Dr Vicky Fraser, in her keynote address at the APIC conference in 2000, reminded us that ICHE programs are cost centers rather than revenue generators, and are often targets for budget cuts. Although Haley's Study on the Efficacy of Nosocomial Infection Control (SENIC), published in 1985, was a landmark event demonstrating the importance of our profession's mission, it is becoming dated. Infection control professionals (ICPs) must continue Haley's work, finding innovative ways to market or demonstrate the value of ICHE programs to health care executives. Closing the 1999 APIC conference with a symposium entitled "Breaking Out of the Box," Jackson and Massanari challenged ICPs to educate themselves about the changing health care environment, to be proactive, and constructively help organizations "re-engineer" more efficiently, rather than feel victimized and helplessly await being re-engineered out of existence. The threat of downsizing prompted ICPs at BJC HealthCare to realize that the time had come to change their own culture and attitudes and to focus on the business of infection control. This change required challenging the traditional roles of solo practitioner, data collector, and keeper of infection control data and knowledge. The goals now include leading intervention teams committed to reducing health care-associated infections, partnering rather than accepting sole responsibility for lowering infection rates, and learning to influence without authority. Staying focused on quality and cost-effectiveness and demonstrating improvements in clinical outcomes became a commitment. This article discusses BJC HealthCare's journey through change so that it may provide useful information and tools for ICPs in any setting looking for the necessary change strategies that might keep them in business. PMID- 11944004 TI - Performance of latex and nonlatex medical examination gloves during simulated use. AB - BACKGROUND: In response to the rise in latex allergies, gloves made from a variety of nonlatex materials have been introduced into the health care environment. To date, at least 1 study, by Rego and Roley (1999), has reported that both latex and nitrile medical examination gloves provide comparable barrier protective qualities. The purpose of our study was to determine the effects of glove stress, type of material (vinyl, nitrile, copolymer, latex), and manufacturer on the barrier effectiveness of medical examination gloves. METHOD: A total of 5510 medical examination gloves (1464 nitrile, 1052 latex, 1006 copolymer, and 1988 vinyl) were divided into 2 groups: stressed and unstressed. Unstressed gloves were visually inspected and water-tested according to the Food and Drug Administration water-testing standards. Stressed gloves were manipulated according to a designated stress protocol, visually inspected, and then subjected to the same Food and Drug Administration water-testing standards. RESULTS: Our limited sample size demonstrated that nitrile gloves had the lowest failure rate (1.3%), followed by latex (2.2%); vinyl and copolymer gloves had the highest failure rate (both 8.2%). With use of a logistic regression analysis adjusting for manufacturer and stress, latex examination gloves were found to be 3 times more likely to fail than nitrile gloves (odds ratio, 3.2; 95% CI, 1.37-7.50). Nitrile gloves were also found to fail significantly less often than vinyl or copolymer gloves (odds ratio, 12.60; 95% CI, 5.80-27.40). CONCLUSIONS: Nitrile examination gloves are a suitable alternative to latex, whereas vinyl and copolymer examination gloves were found to be less effective barriers. Further research is indicated to determine whether nitrile gloves can provide effective barrier qualities during clinical use versus laboratory simulations. PMID- 11944005 TI - Quality assessment and quality control of echocardiographic performance in a large multicenter international study: Valsartan in heart failure trial (Val HeFT). AB - OBJECTIVE: To qualify 302 multinational echocardiography sites to record and read serial studies and to monitor quality in 5010 patients randomized into Valsartan in Heart Failure Trial (Val-HeFT). BACKGROUND: Decentralized echocardiography reading is unprecedented in large clinical trials. METHODS: Single and duplicate recordings, and triplicate readings of echocardiographic variables were submitted to 3 core laboratories. Quality of recording was defined with a 16-point scoring system; accuracy of reading by agreement with core readings; reproducibility by agreement between the duplicate studies. RESULTS: Seventy-five percent of initial submissions were approved for recording, and 50% for reading. Resubmissions were evaluated until approval. Initial scores of sites approved with 1 versus 2 submissions differed, 13.8 +/- 1.4 versus 10.6 +/- 2.0, P <.001; final score was similar, 13.4 +/- 1.6, P = ns. Initial score of sites approved after 3 or more submissions differed, 9.5 +/- 1.9, P <.001; final score improved to 12.7 +/- 1.9, but remained lower, P <.001. Expressed as 95% limits of agreement (mean difference +/- 1.96 x SD), accuracy = -0.04 +/- 0.74 cm for left ventricular internal end-diastolic diameter (LVIDd); -0.29 +/- 14.3% for ejection fraction (EF); reproducibility = 0.00 +/- 0.53 cm for LVIDd; -0.25 +/- 8.3% for EF. Quality of random sampling at baseline, 4, 12, and 18 months showed recording scores of 11.7 +/- 2.7, 12.3 +/- 2.4, 12.1 +/- 2.2, and 11.4 +/- 2.0, P =.24. Power analysis revealed differences of 0.09 cm for LVIDd, and 0.86% for EF detectable with a power of 90% and alpha of 5%. CONCLUSION: The qualifying process improved echocardiography recording and reading to bring 95% of the sites to an equivalent level of quality. Monitoring quality found that recording quality and reading accuracy were maintained 18 months into the trial. Reproducibility, given the large sample size, will be able to detect small changes in LVIDd and EF. PMID- 11944006 TI - Pulse inversion harmonic imaging improves endocardial border visualization in two dimensional images: comparison with harmonic imaging. AB - Pulse inversion harmonic imaging (PIHI) is a new modality that increases the detection of harmonic echoes and myocardial contrast by cancelling linearly transmitted signals. We tested whether PIHI improved the detection of endocardial borders in noncontrast 2-dimensional echocardiography. We compared PIHI with tissue harmonic imaging (THI), which decreases linearly transmitted signals using filters. Fundamental mode (FM) was compared with THI and PIHI in 50 consecutive patients. The global and segmental endocardial visualization scores measured with FM were significantly improved by using either THI or PIHI. The improvement of the global score compared with FM was slightly higher using PIHI than THI, because of an improved visualization of the base and the anterior wall with the PIHI technique compared with THI. The ratio of myocardial-to-cavity signal was similarly increased from FM with THI and PIHI. PIHI, a new modality for detection of myocardial contrast, can also be used for endocardial border visualization. It provides an improvement relative to THI for specific regions of the endocardium. PMID- 11944007 TI - Agitated colloid is superior to saline and equivalent to levovist in enhancing tricuspid regurgitation Doppler envelope and in the opacification of right heart chambers: a quantitative, qualitative, and cost-effectiveness study. AB - BACKGROUND: Doppler spectrum of tricuspid regurgitation (TR) is used to noninvasively assess right ventricular (RV) pressure. With mild TR, the native (Nat) TR envelope may not allow accurate pressure evaluation. Proprietary contrast agents, such as Levovist (Lev) can be used to augment TR Doppler and opacify right-sided heart chambers, but they are expensive, and their efficacy has not been objectively evaluated in patients with difficult baseline studies or compared with less expensive saline (Sal) or colloid solutions, such as Gelofusine (Gel). METHODS: Twenty-five consecutive patients with poor quality Nat TR envelopes on transthoracic echocardiogram were reexamined after serial intravenous injection of 3 contrast agents (Sal, Gel, and Lev). Doppler signals for each agent recorded on video and digitally on optical disk were assessed for signal quality, estimated RV pressure, interobserver and intraobserver variation, and longevity of signal. Quality of right ventricular-right atrial (RV-RA) opacification was also determined for Sal and Gel. Of the 25 patients, 9 underwent percutaneous right-sided heart catheterization. We used the pressures obtained from the catheterization to independently evaluate the pressure estimates from echocardiography. RESULTS: All 3 contrast agents significantly improved the mean quality grade (grades 0-5) of TR envelopes (Nat 1.12, Sal 1.97, Gel 2.56, Lev 2.41, P <.001), decreased the number of uninterpretable envelopes (grade 0) (Nat 49%, Sal 12%, Gel 4%, Lev 12%, P <.0001 for comparison of each agent relative to Nat), and improved the correlation between echocardiographic and catheter-derived RV-RA pressure measurements (Nat r = 0.65, Sal r = 0.75, Gel r = 0.90, Lev r = 0.88). The persistence of enhanced Doppler signals of interpretable quality (> grade 1) was greater for Lev (15.8 seconds) and Gel (15 seconds) than Sal (7.6 seconds) (P =.002). Opacification of RV and RA, measured as mean luminosity score during 2-dimensional harmonic imaging, was significantly higher for Gel than Sal (92.84 +/- 31.2 vs 56.06 +/- 25.6, respectively; P =.0003). Sal, Gel and Lev, respectively, cost $0.10, $2.50, and $75.00 per study. CONCLUSION: Agitated colloid is a novel, effective, and inexpensive alternative to proprietary agents and saline for the assessment of pulmonary systolic pressure and right-sided heart opacification. PMID- 11944009 TI - Spontaneous echocardiographic microbubbles associated with prosthetic mitral valves: mechanistic insights from thrombolytic treatment results. AB - The purpose of this study was to determine the prevalence of microbubbles (MBs) in patients with prosthetic mitral valves (PMVs). The clinical and echocardiographic predictors of MB were investigated. We also analyzed the temporal relation between MBs and the thrombolytic treatment of thrombotic PMV dysfunction. The study material comprised 307 transesophageal echocardiography examinations in 279 patients (170 women and 109 men with a mean age of 37.9 +/- 13.3 years) with PMV. The PMV was mechanical in 245 patients (tilting-disk valves in 129, and bileaflet aortic valves in 116) and bioprosthetic mitral valves in 34 patients. Twenty-eight sessions of thrombolytic treatment were performed because of the obstructive (n = 18) and nonobstructive (n = 10) thrombi involving the PMV. No MBs were seen in any of the bioprosthetic valves. The MBs were present in 128 of 227 (56.4%) PMV without obstruction compared with only 1 of 18 (5.5%) valves with thrombotic obstruction (P <.0001). The MB were documented in 75.4% of the normal bileaflet valves compared with 38.5% of the tilting-disk valves (P <.0001). The MB intensity score was also significantly higher in the bileaflet valves (2.0 +/- 0.8 vs 0.7 +/- 0.7, P <.05). The incidence of MBs increased from 5.5% to 68.7% after successful thrombolysis in patients with obstructive PMV thrombi (P <.001). There were no other predictors of MBs in this series. The passage of MBs in the aortic root was not documented in any instances. We conclude that MBs are normal echocardiographic findings depending on the type and function of the mechanical PMVs. PMID- 11944008 TI - Quantitative diagnosis of apical cardiomyopathy using contrast echocardiography. AB - BACKGROUND: The echocardiographic diagnosis of apical hypertrophic cardiomyopathy (ACM) has been limited by the frequent inability to visualize the apical endocardium. We hypothesized that the use of contrast agents in patients with suspected ACM, but nondiagnostic echocardiographic studies, would allow quantitative diagnosis. METHODS: Contrast enhancement was performed in 26 patients with nondiagnostic transthoracic echocardiograms (TTEs) for the diagnosis of ACM; 6 patients with suspected ACM based on unexplained symmetric precordial T-wave inversions and increased apical tracer uptake on single-photon emission computed tomography (SPECT) scans, 10 patients with normal electrocardiogram (ECG) readings and no history of hypertension (healthy group), and 10 patients with hypertension and ECG criteria for left ventricular hypertrophy (LVH group). Images were obtained with Optison (Mallinckrodt Medical; IV, 1.0 mL) using harmonic imaging and low mechanical index. Posterior (PW) and septal wall (SW) thicknesses were measured at end-diastole in the parasternal long-axis view. Apical wall thickness (A) was measured from the contrast-enhanced apical endocardium to the visceral epicardial surface in the apical 4-chamber view. A/PW and A/SW ratios were calculated for each group. Asymmetric apical hypertrophy was defined as an A/PW ratio greater than 1.5. RESULTS: Contrast enhanced apical thickness was greater than 2.0 cm in all patients in the suspected ACM group but less than 1.2 cm in all patients in the LVH and healthy groups. In all 6 patients in the suspected ACM group, A/PW and A/SW ratios were greater than 1.5. No patient in the healthy or LVH groups had thickness ratios greater than 0.85. CONCLUSION: Contrast echocardiography allows quantitative diagnosis of ACM in patients with suggestive ECG and SPECT but nondiagnostic TTEs. This study suggests that contrast echocardiography should be performed before using more expensive or invasive diagnostic testing for this condition. PMID- 11944010 TI - Blood flow velocity profiles in the aortic annulus: a 3-dimensional freehand color flow Doppler imaging study. AB - BACKGROUND: The use of a single sample volume in Doppler measurements of the velocity time integral (VTI) in the aortic annulus may introduce errors in calculations of stroke volumes, shunts, regurgitant fractions, and aortic valve area. To study the blood flow velocity distribution and assess this potential error, we used a dynamic 3-dimensional color flow Doppler imaging method. METHODS AND RESULTS: Seventeen healthy volunteers were studied. The ultrasound data were captured from 10 to 20 heartbeats at a high frame rate (mean 57 frames per second) while freely tilting the transducer in the apical position. A magnetic position-sensor system recorded the spatial position and orientation of the probe. The raw digital ultrasound data were analyzed off-line with no loss of temporal resolution. Blood flow velocities were integrated across a spherical surface that tracked the aortic annulus during systole. The ratios of the systolic maximum to the systolic mean VTI ranged from 1.2 to 1.5 (mean 1.4). At the time of systolic peak flow, the ratios of the maximum to the mean velocity ranged from 1.1 to 2.0 (mean 1.5). The location of the maximum velocities and VTI showed individual variation. CONCLUSION: The blood flow velocity profile was nonuniform. By using a single sample volume in Doppler measurements of the VTI in the aortic annulus, errors ranging from 20% to 50% may be introduced in calculations of stroke volumes. PMID- 11944011 TI - Restrictive left ventricular filling and preserved ventricular function: a limitation in the noninvasive estimation of pulmonary wedge pressure by Doppler echocardiography. AB - Pulmonary wedge pressure (PWP) can be estimated from the ratio of transmitral early peak flow velocity to flow propagation velocity measured by Doppler. Discrepancies observed in the application of the method prompted us to design this prospective study, aimed at detecting potential limitations of the method. We studied a total of 32 patients admitted to a cardiac intensive care unit, using a Swan-Ganz catheter. Correlation between invasive- and Doppler-estimated PWP was fairly good (r = 0.58). Analysis of discrepant cases led to the identification of a subgroup of 6 patients in whom PWP was largely underestimated because of unexpectedly high values of flow propagation velocity (71 +/- 15 cm/s vs 37 +/- 10 cm/s in the rest of the group; P =.0001). All of them had in common a restrictive Doppler filling pattern and preserved left ventricular systolic function. Exclusion of this group showed an improvement in the correlation coefficient to r = 0.72. In conclusion, PWP can be estimated by the Doppler early peak flow velocity to flow propagation velocity ratio, although a significant underestimation of PWP may be observed in patients with a restrictive filling pattern and preserved ventricular function. PMID- 11944012 TI - The left ventricular color M-mode Doppler flow propagation velocity V(p): in vivo comparison of alternative methods including physiologic implications. AB - Three alternative methods have been proposed for determining the "velocity of propagation" (V(p)) from color M-mode Doppler transmitral flow images. A quantitative intermethod comparison is performed by using a broad class of images encountered in clinical practice. V(p) for 13 subjects was measured by using each of the 3 methods and then statistically compared. Differences among the definitions of V(p) generated large differences in its numerical value when computed from the same image. Depending on the choice of method, the value of V(p) can vary by as much as 250%. Advantages and limitations of each method are discussed in the context of the physical meaning of V(p). Development of an optimal version of V(p) and consensus on how it should be defined merits further attention. This will require a multidisciplinary approach encompassing physiology, fluid dynamics, image processing, and mathematical modeling. PMID- 11944013 TI - Anatomic and hemodynamic imaging using a new vector phased-array intracardiac catheter. AB - BACKGROUND: We used a new vector, phased-array intracardiac catheter (AcuNav) with complete 2-dimensional imaging and Doppler capabilities to describe a systematic approach for a detailed anatomic and hemodynamic cardiac assessment. METHODS: In 14 dogs, the intracardiac echocardiographic catheter was inserted through an 11F venous access and placed in the right side of the heart to perform a comprehensive ultrasound examination of the heart. RESULTS: Imaging was successful in all dogs. All 4 cardiac chambers and valves were imaged clearly in multiple orientations. Additional structures seen included the vena cavae, coronary sinus, right and left appendages, interarterial septum, coronary arteries, and all 4 pulmonary veins. Intra-abdominal structures, such as the aorta, liver, and hepatic veins were also visualized. A complete Doppler examination of intracardiac and paracardiac flows was also possible. CONCLUSION: AcuNav is a unique intracardiac imaging device, which allows comprehensive structural and functional cardiac assessment. PMID- 11944014 TI - Quantification of regional left ventricular wall motion in newborns by color kinesis. AB - BACKGROUND: Normal values for regional left ventricular wall motion, although documented in adults, have not been reported in healthy newborns. METHODS: This study prospectively evaluated global and segmental systolic and diastolic cardiac function by color kinesis in clinically asymptomatic healthy newborns. RESULTS: Eighty-eight asymptomatic infants who were less than 48 hours old were studied. Systolic and diastolic parameters of global and regional left ventricular function are reported as means +/- SD, medians, 5th and 95th percentiles to establish the normative values for newborns. The reported fractional area changes during systole and diastole are similar to the reported normal values for older subjects. Higher body surface area significantly correlated with an increased peak velocity during systole, and fractional area changes during filling of the lateral wall. CONCLUSIONS: Our report of left ventricular regional wall-motion characteristics of healthy newborns, as evaluated by color kinesis, may help in the objective evaluation and management of newborns suspected to have global or segmental ventricular dysfunction. PMID- 11944015 TI - Mechanism of tricuspid regurgitation in paramembranous ventricular septal defect. AB - Literature has been limited in regard to the mechanisms of tricuspid regurgitation (TR) in patients with paramembranous (perimembranous) ventricular septal defect (VSD). Most observations have noted tricuspid valve clefts or dysplasia. We describe another mechanism for production of TR in association with paramembranous VSD. In 8 patients, we found significant TR produced by the VSD jet pushing the tricuspid anterior leaflet forward to open the tricuspid valve orifice. In these patients, a moderate paramembranous VSD extended slightly below the septal tricuspid leaflet with only partial obstruction of the VSD jet. All patients had restrictive VSD with low right ventricular pressure. This mechanism to produce TR was best defined by intraoperative transesophageal echocardiography, but current higher resolution imaging should allow correct diagnosis. We believe that when this mechanism for TR is found in association with a moderate VSD, surgical VSD closure is warranted. PMID- 11944016 TI - Hand-carried cardiac ultrasound (HCU) device: recommendations regarding new technology. A report from the Echocardiography Task Force on New Technology of the Nomenclature and Standards Committee of the American Society of Echocardiography. AB - The newest introduction to echocardiography is a hand-carried ultrasound (HCU) device. It is a small echocardiographic machine that typically weighs less than 6 lb and can obtain echocardiographic images and data. However, neither the device nor the context of the examination fulfills the criteria for a comprehensive or complete echocardiographic examination. The American Society of Echocardiography believes that HCU will extend the concept of the "complete physical examination," allowing more rapid assessment of cardiovascular anatomy, function, and physiology. However, appropriate user-specific training (Level 1 at a minimum) and assumption of responsibility are essential to ensure the most accurate acquisition, interpretation, and use of the data. PMID- 11944017 TI - Diagnosis of myocardial perforation by a Greenfield filter made by transesophageal echocardiography. AB - Greenfield filters have been successfully used to prevent pulmonary embolism in selected patients. However, various complications have been described. A patient is presented in whom a Greenfield filter migrated to the right atrium, and subsequently perforated the right atrial wall. The diagnosis was made by transesophageal echocardiography and the filter removed surgically. This case emphasizes the role of transesophageal echocardiography in the management of migrated Greenfield filters. PMID- 11944018 TI - Quantification of left-to-right shunt in patent ductus arteriosus with the PISA method. AB - Direct Doppler echocardiographic calculation of shunt flow in patent ductus arteriosus (PDA) may be difficult because the orifice area is not well visualized. We used proximal isovelocity surface area (PISA) method to calculate the effective shunt orifice area and the shunt volume in a patient with PDA. The PISA calculation correlated well with shunt estimation by echocardiographic volumetric methods and by oximetry. PMID- 11944019 TI - Early appearance of echo-contrast simulating an intracardiac shunt in a patient with liver cirrhosis and intrapulmonary shunting. AB - Time of appearance of agitated saline contrast in the left atrium is one of the distinguishing features between intracardiac and intrapulmonary shunt. We report a patient with hepatic cirrhosis who had intrapulmonary shunting and had appearance of peripheral venous saline contrast injection in the left heart chambers within 3 cardiac cycles after its appearance in the right heart chambers despite absence of an intracardiac shunt. Thus, time of appearance of saline contrast in the left heart chambers should not be the sole criterion to distinguish intracardiac from intrapulmonary shunt. PMID- 11944020 TI - Papillary fibroelastoma of the aortic valve. AB - We hereby report the case of a patient with a papillary fibroelastoma involving the right coronary cusp of the aortic valve that was diagnosed with the use of transesophageal echocardiography after workup for recurrent transient ischemic attacks. PMID- 11944021 TI - Fistulous communication between the right coronary artery and left atrium. AB - This report describes congenital coronary artery fistula of the right coronary artery draining into the left atrium in a young woman. The initial diagnosis of this rare anomaly was made noninvasively by echocardiography. Transesophageal echocardiography was vastly superior to transthoracic echocardiography in presenting the entire course of the fistula. The diagnosis was confirmed by coronary angiography. The fistula was successfully closed surgically. PMID- 11944022 TI - Lipoprotein (a): where does the atherogenicity reside? PMID- 11944023 TI - Antioxidant nutrient supplementation and brachial reactivity in patients with coronary artery disease. AB - Epidemiologic studies have shown a correlation between antioxidant intake and coronary artery disease (CAD); however, the results of clinical trials have been inconsistent. We evaluated the effect of combined antioxidant supplementation on endothelial function and its correlation with change in low-density lipoprotein cholesterol (LDLC) oxidation in patients with established CAD. In a double-blind, placebo-controlled 12-week trial, 18 nonsmoking, nondiabetic patients (mean age 62.4 +/- 8.1 years) were randomized to receive placebo or antioxidant supplementation consisting of (a) 400 IU of vitamin E, 500 mg of vitamin C, and 12 mg of beta-carotene; or (b) 800 IU of vitamin E, 1000 mg of vitamin C, and 24 mg of beta-carotene daily. Endothelial function was evaluated on the basis of percent and absolute changes in brachial artery diameter in response to reactive hyperemia induced by occlusion-release. Baseline and 12-week values of LDL oxidation (measured on the basis of lag phase), endothelial function, dietary composition, serum antioxidants, and lipids were measured. We noted a significant between-group difference at 12 weeks for change in plasma concentrations of alpha tocopherol, vitamin C, and beta-carotene between the placebo and antioxidant groups (p <.05). Both placebo and treatment groups demonstrated a significant improvement in lag phase; however, the treatment group achieved a greater, although nonsignificant, magnitude of change compared with the placebo group (181.3 +/- 177.8 minutes vs 80.6 +/- 63.0 minutes, P =.06). Within-group change in brachial reactivity from baseline to follow-up in the treatment group did not reach statistical significance (1.7% +/- 3.2% and 0.07 mm +/- 0.13 mm, P =.08 and P =.09, respectively), whereas an improved change in brachial reactivity was observed in the placebo group (2.2% +/- 1.9%, 0.09 mm +/- 0.06 mm, P <.05). No significant correlation was found between change in lag phase and change in endothelial function. On adjustment for confounders, antioxidant supplementation was found not to be a significant predictor of brachial reactivity. We conclude that antioxidant supplementation did not significantly alter brachial reactivity, despite significantly increased plasma levels of antioxidants and improved lag phase. These data should be confirmed in larger-scale trials and examined in studies evaluating individual dietary antioxidant supplementation. PMID- 11944024 TI - Effects of TNF-alpha on glucose metabolism and lipolysis in adipose tissue and isolated fat-cell preparations. AB - We hypothesized that exposure to tumor necrosis factor-alpha (TNF-alpha) would significantly increase lactate production by adipose-tissue (AT) fragments and isolated adipocytes. We therefore examined the effects of TNF-alpha on the metabolism of epididymal AT explants during 24-hour tissue incubation. We also studied the effects of this 24-hour TNF-alpha tissue exposure on subsequent glucose metabolism and lipolysis by isolated adipocytes. Glycerol release into the medium was significantly increased (50%, P =.027) by exposure of the AT fragments to TNF-alpha (4 nmol/L) for 24 hours. During this time, glucose uptake from the medium and lactate release into the medium tended to increase, whereas leptin release into the medium tended to decrease, but these effects of TNF-alpha were not statistically significant. After the 24-hour AT-explant incubation, adipocytes were isolated by means of collagenase digestion from the AT fragments and subsequently tested in a short-term (60-minute) metabolic incubation. Prior exposure to TNF-alpha resulted in a significant increase in adipocyte glycerol release (P =.044), total glucose metabolism (P =.019), and lactate production (P =.037). With the exception of lactate, TNF-alpha produced no significant stimulation of the metabolites of glucose. The pattern of glucose metabolism elicited by TNF-alpha exposure differs from that usually attributed to a lipolytic hormone and suggests that the effects of TNF-alpha on glucose metabolism involve pathways separate from, or in addition to, its effects on lipolytic stimulation. PMID- 11944025 TI - Antiphospholipid antibody syndrome: rapid, sensitive, and specific flow cytometric assay for determination of anti-platelet phospholipid autoantibodies. AB - A rapid, sensitive and specific flow cytometric assay has been developed for the determination of autoantibodies directed against platelet anionic phospholipids in antiphospholipid antibody syndrome (APLAS). The method is based on demonstrable competition between the placental anticoagulant protein I, annexin V, and the patients' autoantibodies on the platelet anionic phospholipids (the binding site for the prothrombinase complex; prothrombin and factors Va and Xa). The method is practical and rapid, uses readily available reagents, and involves standard equipment. The assay is inexpensive and cost-effective for both single and multiple samples. Results are provided within 2 hours from obtaining blood samples, thereby supporting clinical decision-making and management. Ten serum samples from patients with the clinical diagnosis of APLAS (48 tests), 10 from normal individuals (35 tests), and 10 from patients with immune thrombocytopenia (33 tests) were tested. Platelet preparations preincubated with normal sera showed high binding of fluorescein-labeled annexin V with an average fluorescence of 202.9 +/- 22.0 (arbitrary units). Patients with immune thrombocytopenia exhibited similar results, with an average fluorescence of 192.5 +/- 32.1 (P >.05). In contrast, incubation with sera from patients with APLAS resulted in a marked decline in the binding of annexin V to an average fluorescence of only 14.6 +/- 7.4 (P <.001). Preincubation with annexin V followed by the addition of patients' sera showed displacement of annexin V to a similar degree. Because annexin V attenuates procoagulant activity by competing with factors Va and Xa on the platelet anionic phospholipids, its displacement by patients' antibodies may result in the acceleration of procoagulant activity, thereby promoting thrombogenesis in APLAS. PMID- 11944026 TI - Extraintestinal pathogenic Escherichia coli: "the other bad E coli". AB - Extraintestinal pathogenic Escherichia coli (ExPEC), the specialized strains of E coli that cause most extraintestinal E coli infections, represent a major but little-appreciated health threat. Although the reasons for their evolution remain mysterious, by virtue of their numerous virulence traits ExPEC clearly possess a unique ability to cause disease outside the host intestinal tract. Broader appreciation of the existence and importance of ExPEC and better understandings of their distinctive virulence mechanisms, reservoirs, and transmission pathways may lead to effective preventive interventions against the morbid and costly infections ExPEC cause. PMID- 11944027 TI - GpIIb/IIIa is the main receptor for initial platelet adhesion to glass and titanium surfaces in contact with whole blood. AB - Platelets are the first cells to adhere to a surface in contact with blood and are capable of mediating several different responses after contact with different protein-coated surfaces. They are the main source of growth factors such as platelet-derived growth factor and are therefore important in the healing process. In this study, initial platelet adhesion to and spread on hydrophilic and hydrophobic (methylized) glass and titanium with similar wettability were investigated. Whole coagulating blood was used to simulate the in vivo situation shortly after implantation, in which bleeding precedes inflammation and wound healing. Several different antibodies directed against platelet integrins and receptors (CD9, FcgammaRII, GPIIb/IIIa, vitronectin receptor, GPIb/V/IX) were used in an attempt to block platelet adhesion to the surfaces. Immunofluorescence results show that initial platelet adhesion to all the surfaces we investigated can be almost completely inhibited (approximately 95%) by clone M148, an antibody against the GPIIb/IIIa complex (integrin alpha(IIb)beta(3); CD41/CD61), but not with other antibodies to the separate parts of the integrin. Antibodies known to inhibit fibrinogen binding to GPIIb/IIIa after adenosine diphosphate- and collagen- induced aggregation had very little effect on initial platelet adhesion. None of the other integrins were found to have such an effect on initial platelet adhesion. Antibody clone M148 was furthermore found to inhibit platelet spreading. This study shows that regardless of wettability and the biomaterial used, initial adhesion of platelets appears to be mediated by GPIIb/IIIa binding to surface adsorbed fibrinogen. PMID- 11944028 TI - Nasal lavage as a tool for the assessment of upper-airway inflammation in adults and children. AB - The prevalence of respiratory allergies has increased over the last 20 years, highlighting the need for a simple and noninvasive tool to investigate, in a clinical and epidemiological context, airway-inflammation mechanisms encountered in allergic and inflammatory processes. The nose, as the first region of the respiratory tract to come in contact with airborne pollutants, is easily explored with the use of nasal lavage (NL). We evaluated an NL method for adults and children, along with its reproducibility and capacity to separate different subgroups. NL reproducibility, assessed in 10 healthy, nonsmoking adults on three different occasions, was determined with the use of the intraclass coefficient of correlation for such inflammatory markers as total cell count, albumin, urea, neutrophil elastase, alpha(1)-antitrypsin, interleukin-6, and interleukin-8. Using this NL method, we analyzed nasal markers of 50 healthy adults (smokers and nonsmokers) and 12 healthy children. Our NL method demonstrated high reproducibility with regard to total cell count, albumin, urea, and alpha(1) antitrypsin (intraclass correlation coefficient > 0.75). Compared with NL results in nonsmokers, NL in heavy smokers revealed significant increased concentrations of total cell counts and interleukin-8 and significant decreased concentrations of interleukin-6. These findings suggest that NL can be used as a tool in the assessment of inflammation because it has the correct reproducibility and can discriminate between heavy smokers and nonsmokers. Moreover, the use of this standardized method in children is feasible. PMID- 11944029 TI - Heterozygous apolipoprotein (a) status and protein expression as a risk factor for premature coronary heart disease. AB - Exactly how apolipoprotein a [APO(a)] isoform size affects the degree of cardiovascular risk associated with high lipoprotein a [LP(a)] levels is not fully understood. Using a sodium dodecyl sulfate-agarose APO(a) & LP(a) phenotyping method, we assessed the role of APO(a) size heterogeneity according to the number of kringle 4 repeats and the differential APO(a) protein expression in 91 male Spanish patients with premature coronary heart disease (CHD) compared with 99 healthy Spanish men. CHD patients had significantly increased median plasma LP(a) levels (0.31 g/L) and a higher percentage of subjects with LP(a) levels of 0.30 g/L or greater (51%) than controls (0.15 g/L and 23%, respectively). Patients with the double-band phenotype had significantly higher plasma LP(a) levels (median 0.37 g/L) compared with those expressing a single band phenotype (median 0.20 g/L; P =.018) and with their corresponding controls (median 0.15 g/L; P <.001). The double-band phenotype and LP(a) values of 0.30 g/L or greater had a significant association with CHD (odds ratio [OR] 6.47, 95% confidence interval [CI] 2.51-16.7), stronger than that observed for the entire group (OR 4.19, 95% CI 1.97-8.90). The adjusted OR for the APO(a) protein pattern that equally expressed both isoforms indicates an independent association with premature CHD (OR 3.33; 95% CI 1.08-10.3). These results suggest that APO(a) phenotyping might be used in subjects with hyperlipoproteinemia a as a powerful marker to assess the risk of premature CHD because heterozygous status, mainly when both isoforms are equally expressed, is associated with higher cardiovascular risk. PMID- 11944032 TI - Networks and crosstalk: integrin signalling spreads. AB - Multicellular organisms must coordinate signals from adhesion receptors with those from other signalling receptors (for example, growth factor receptors). Here, we briefly review paradigms of integrin-adhesion-receptor signalling. We discuss how adhesive signalling is coordinately regulated through intersecting networks. We also examine some examples of how some forms of integrin crosstalk may lead to unforeseen and potentially deleterious responses. PMID- 11944033 TI - Herpesvirus hijacks an integrin. PMID- 11944034 TI - Neighbourhood Watch for the cytoskeleton. PMID- 11944035 TI - E-cadherin and Hakai: signalling, remodeling or destruction? PMID- 11944036 TI - Ready and abl. PMID- 11944038 TI - How Rab proteins link motors to membranes. PMID- 11944037 TI - Integrin regulation of growth factor receptors. PMID- 11944039 TI - A double-edged sword. PMID- 11944041 TI - Sensing the environment: a historical perspective on integrin signal transduction. AB - Cell adhesion mediated by integrin receptors has a critical function in organizing cells in tissues and in guiding haematopoietic cells to their sites of action. However, integrin adhesion receptors have broader functions in regulating cell behaviour through their ability to transduce bi-directional signals into and out of the cell and to engage in reciprocal interactions with other cellular receptors. This historical perspective traces the key findings that have led to our current understanding of these important functions of integrins. PMID- 11944042 TI - Microscope-based techniques to study cell adhesion and migration. AB - Modern light microscopy has evolved to provide a variety of quantitative imaging techniques and also the capability to perturb structure-function relationships in living cells. The advances have been especially useful in the study of cell adhesion and migration. This review will focus on how such microscopy-based techniques can be useful in situ to study the molecular interactions and dynamics, to locally perturb actin-based structures and to measure the traction forces exerted by motile cells. PMID- 11944043 TI - Adhesion assembly, disassembly and turnover in migrating cells -- over and over and over again. AB - Cell migration is an integrated process that requires the continuous, coordinated formation and disassembly of adhesions. These processes are complex and require a regulated interaction of numerous molecules, and the activation of specific signalling pathways. Even though understanding these processes is challenging, important insights are beginning to emerge, and the technology to facilitate significant advances in this area is now in place. PMID- 11944044 TI - Intercellular adhesion, signalling and the cytoskeleton. AB - Connections between the cytoskeleton and intercellular junctions profoundly influence cell shape and motility. It is becoming increasingly clear that in addition to structural functions, components of the adhesion apparatus also possess signalling capabilities. Recent studies suggest that their dual function may provide the means to integrate changes in morphology and gene expression during tissue and organ development. PMID- 11944046 TI - A useful protocol for in situ RT-PCR on plant tissues. AB - This study describes an effective method of in situ RT-PCR (RT-ISPCR) that was developed to localize gene expression in plant tissues. This RT-PCR technique was performed on sectioned tissues of female buds of the cucumber GY3 inbred line. The CUS1 gene, encoding the MADS-box type (agamus-like) protein, the expression pattern of which was described earlier, was used as a marker gene for optimisation of steps in the in situ RT-PCR inside the cells. For the identification of RT-PCR products inside the cells of the female buds, they were fixed in FAA solution, embedded in Paraplast Plus and cut into 7 microm thick sections which were dewaxed by immersion in HistoClear and dehydrated with ethanol. They were washed in water, then in 0.02M HCl, 2xSSC and PBS buffer. In the next step of tissue pretreatment, the sections were digested with 1% pectinase. As shown, the pectinase treatment proved to be a crucial step in the tissue preparation procedure to get successful RT-PCR products. After washing in PBS buffer, the sections were digested with protease K followed by incubation with RNase-free DNase I, and subsequently washed in 2xSSC, 1xSSC and 0.5xSSC and finally in DEPC-treated water. Then the sections were covered with 50 microl of the RT-PCR reaction mixture supplemented with 0.5 microM digoxigenin dUTP and sealed with a coverslip. After amplification in situ the PCR products were identified with anti-digoxigenin antibody (Roche Molecular Biochemicals), conjugated with alkaline phosphatase. The data obtained showed that specific signals reflecting CUS1 gene expression were detected in the female flower buds of cucumber. The specificity of the in situ RT-PCR protocol was confirmed by dot blot hybridization of RT-PCR products with CUS1 cDNA probe. PMID- 11944047 TI - The production of transgenic potato plants expressing human alpha-interferon using lipofectin-mediated transformation. AB - The lipofectin system was used to transform potato protoplasts with plasmid DNA (pIG3031), which contains human alpha-interferon cDNA and codes for the selectable neomycin phosphotransferase II gene (NPT II). Both genes are under the control of the bi-directional plant active transcriptional promoter from Agrobacterium tumefaciens T-DNA. The criteria of phenotype stability after selective pressure removal, in vitro activity assay of NPT II, the biological analysis of alpha-interferon activity, Southern blot analysis and RNA slot blot were used to confirm the mitotic stability of the foreign gene and its expression and stable integration into the host genome. These studies demonstrate that human alpha-interferon cDNA can be correctly expressed in potato cells. PMID- 11944048 TI - The mechanism of the hemolytic activity of polyene antibiotics. AB - The kinetics of the filipin-, amphotericin B- and nystatin-induced hemolysis of human erythrocytes were investigated. Filipin-induced hemolysis is of the damage type. It is an all-or-none process, partly inhibited by Ca2+ or Ba2+ but not by Mg2+, Na+ or SO42-. The hemolytic activity of filipin is explained by the formation of large aggregates within the erythrocyte membrane in the form of large perforations, permeable to substances of low molecular weight as well as to macromolecules, including hemoglobin. In isotonic KCl solution, both amphotericin B and nystatin, at low concentrations, form smaller aggregates within the membranes. As a result, the permeability of the membranes to KCl increases and hemolysis occurs. However, the kinetics of the hemolysis induced by the two polyenes is complex. The process shows some features of the permeability type and some of the damage type. It is suggested that amphotericin B and nystatin may simultaneously form a number of transport systems, differing in their molecular organisation and hemolytic activity. Their participation in erythrocyte membrane permeability can be modified by small changes in membrane organisation and the chemical composition of the incubation medium. In isotonic solutions of divalent cation chlorides, and at higher antibiotic concentration, additional aggregates, allowing divalent cations to permeate, appear. These structures do not permit SO4(2-) to permeate. PMID- 11944049 TI - Stable genetic transformation of garlic plants using particle bombardment. AB - The improvement of garlic plants (Allium sativum L.) via biotechnological approaches is currently limited by the lack of an applicable direct gene transfer system. In this paper, we present the development of a genetic transformation system using particle bombardment for gene delivery and immature clove-derived callus as the gene target. Plasmid DNA (pBI221.23), containing the selectable "hpt" gene for hygromycin resistance and the reporter "gus" gene, was delivered into callus tissue that had been previously treated with aurintricarboxylic acid as an endogenous nuclease inhibitor. The transformed calli were selected using hygromycin B, regenerated, and analysed at the molecular level using DNA hybridization, transgenome rescue and histochemical beta-glucuronidase assay. The results indicated that biolistic transformation can lead to the transfer, expression and stable integration of a DNA fragment into garlic chromosomal DNA. The relative simplicity of this system is a good recommendation for its future use in the production of genetically modified garlic plants. PMID- 11944050 TI - Fatty acid profiling: its usefulness in the evaluation of microbial associations with the green microalga Apatococcus constipatus. AB - To determine differences in microbial community structures, fatty acids from two strains of the green microalga Apatococcus constipatus were isolated and identified by instrumental means. The main fatty acids found were 16:0 and 14:0. These predominant acids represented more than 53% of the total fatty acid (content in both algal isolates. In addition, saturated fatty acids were present in much greater quantity than unsaturated ones. Differences between the strains in the composition of other, modified fatty acids were also evident. The occurrence of fatty acid biomarkers characteristic of certain taxonomic groups confirmed the presence of Gram-positive and Gram-negative bacteria, and fungi. Those observed variations were undoubtedly due to distinct community structures of symbiotic microorganisms living in close associations with the alga. The results presented here indicate that different isolates of the same alga might exhibit different microbial community structures. PMID- 11944051 TI - The biomechanics of standing and balancing in paralyzed people. AB - Prolonged immobilization results in several physiological problems. It has been demonstrated that standing exercises can ameliorate many of these problems. Standing exercises can be performed efficiently with the help of functional electrical stimulation (FES). A novel robotic mechanism which aids the unsupported standing of paraplegics, providing balancing exercise has been developed. The balancing strategy is based on voluntary activity of the paraplegic's upper body and artificially controlled stiffness in the ankles. PMID- 11944052 TI - Some multiple-time-scale problems in molecular dynamics. AB - Many physical problems, particularly in chemical and biological systems, involve processes that occur over widely varying time scales. Such problems have motivated the development of new methods for treating multiple-time-scale problems in molecular dynamics (MD). Methods have been developed for determining the vibrational frequencies and normal modes of large systems in full and reduced conformational space. A method is given for quasiharmonic analysis and reduced quasiharmonic analysis. PMID- 11944053 TI - Magnetic resonance imaging of arterial thrombi and its possible correlation to fibrinolytic treament. AB - For 13 patients with subacute and 4 patients with chronic occlusion, magnetic resonance imaging (MRI) of occlusive arterial thrombi in the superficial femoral artery were performed in vivo. The patients with subacute occlusion were treated with catheter-directed thrombolysis. The frequency of MR signal intensity and its distribution in thrombi were studied for 11 successfully and 2 unsuccessfully treated patients and patients with chronic occlusion. Intra-arterial thrombi were MRI inhomogenous in all of the patients, but the MR signals from lysable and chronic thrombi were significantly different than those from nonlysable ones. The MRI of occlusive arterial thrombi is probably usable to predict the therapeutic outcome of thrombolytic treatment. PMID- 11944054 TI - Structural studies on membrane proteins using non-linear spin label EPR spectroscopy. AB - Non-linear electron spin resonance (EPR) techniques suitable for measuring proximity relationships in membranes are reviewed. These were developed during the past decade in order to measure changes sensitively in the spin-lattice relaxation time (T1) of nitroxyl spin labels covalently attached to membrane lipids or proteins. In combination with paramagnetic quenching agents and double spin-labelling, the methods were further developed for distance measurements. Selected examples are given to illustrate different methods, and types of data obtained for both integral and peripheral membrane proteins. PMID- 11944055 TI - Membrane destabilizations supporting electropermeabilization. AB - The cell membrane can be permeabilized when subjected to calibrated short electric pulses. This membrane alteration can be reversible, leaving cell viability unaffected. This set of events is called Electropermeabilization. This is now used in clinical applications to introduce hydrophilic drugs into the cytoplasm. While an empirical control of the electrical parameters is obtained, our knowledge of the molecular mechanisms supporting the membrane alteration is very limited. This paper gives a critical review of the limits of the models which are proposed by taking into account the different experimental approaches. PMID- 11944056 TI - A generalized mathematical model of biological oscillators. AB - Biological rhythms such as circadian rhythms, biochemical rhythms and neural oscillators are based on the mathematical model of the theory of harmonic oscillators. These are solutions of certain second-order differential equations. They can also be viewed as spherical harmonics on the circle in the two dimensional Euclidean space. The spherical harmonics on (n-1)-spheres and, more generally, the Stiefel harmonics can represent oscillatory phenomena, and we expect that they can serve as models for more complex biological rhythms. PMID- 11944057 TI - Intravascular plug formation induced by poly-APS is the principal mechanism of the toxin's lethality in rats/rat tissues. AB - Toxic water soluble polymeric 3-alkylpyridinium salts isolated from the sponge Raniera sarai strongly inhibited AChE in vitro. In vivo, experimental animals died due to plugs formed in microcirculation. The mechanism of this plug formation is unknown. In vitro, the toxin did not affect the coagulation rate, but the rate of platelet aggregation was accelerated in a dose-dependent manner. The hemolytic activity of poly-APS was diminished by the addition of serum proteins in a dose-dependent manner. These results support the conclusion that non-specific binding to proteins is the underlying mechanism of the lethality of poly APS. PMID- 11944058 TI - The influence of the biomechanical parameters of the hip on the outcome of treatment of hips subject to avascular necrosis of the femoral head. AB - We were interested in whether or not the biomechanical status of the hip influences the course of avascular necrosis of the femoral head. To investigate this, we used a computer aided system based on a three dimensional mathematical model for determining the stress distribution in the hip joint from standard anteroposterior rentgenographs (X-ray images) of both hips and pelvis. Based on the results of our study, we suggest that the biomechanical parameters of the hip play an important role in the outcome of treatment of hips affected by avascular necrosis of the femoral head. PMID- 11944059 TI - Examination of fragment dose contribution in heavy ion radiotherapy. AB - Heavy-ion radiotherapy is an efficient method for the treatment of deep-seated tumors, because the stopping of ions in a tissue delivers the maximal absorbed dose to the tumor-affected areas with minimal damage to the healthy tissues. However, heavy ions can undergo nuclear reactions, giving products with lower Z values and hence a longer range in the tissue. This causes a dose increase beyond the mean range of the primary beam. The contribution of such reaction products was examined in an experiment where a stack of tissue-like targets interleaved with CR-39 etched track detectors (ETD) was irradiated with heavy ions. The analysis was performed using a recently developed technique of trajectory tracing, which enables the spectroscopy of fragments with different Z-values. PMID- 11944060 TI - Frequency encoding of intracellular Ca2+ signals. AB - In a very recent theoretical study, we showed that simple mitochondrial kinetics, originally proposed by Marhl et al., could be easily used as a plug-in element in other models describing intracellular Ca2+ dynamics. We analysed several previously published models and showed that mitochondria could indeed maintain the constant amplitude of intracellular Ca2+ oscillations very effectively. We also pointed out the importance of amplitude regulation for the frequency encoding of intracellular Ca2+ signals. This paper focuses on giving a more exhaustive demonstration of this phenomenon of frequency encoding for the model of Dupont et al. PMID- 11944061 TI - CROW for large scale macromolecular simulations. AB - CROW (Columns and Rows Of Workstations - http://www.sicmm.org/crow/) is a parallel computer cluster based on the Beowulf (http://www.beowulf.org/) idea, modified to support a larger number of processors. Its architecture is based on point-to-point network architecture, which does not require the use of any network switching equipment in the system. Thus, the cost is lower, and there is no degradation in network performance even for a larger number of processors. PMID- 11944062 TI - Study of water migration by micro MR imaging. AB - Magnetic Resonance (MR) microscopy is a noninvasive tool that enables high resolution imaging of water concentration in non-magnetic materials. In this study, water migration in a dense porous material (dental cement) was monitored by 1D MR microscopy. The measured concentration profiles were later analyzed via three different migration models: an imbibition model, a diffusion model and a combined model. From the best fit to the experimental data, model parameters were obtained. PMID- 11944063 TI - Cyclophosphamide metabolites perforate the apical plasma membranes of urothelial cells in rats. AB - The apical plasma membrane of differentiated superficial urothelial cells is characterised by the presence of asymmetric unit membrane (AUM). Cyclophosphamide (CP) metabolites cause perforation of these thickened membranes. In this study, apical plasma membranes were examined after CP injection by electron microscopy. The immediate effect of the CP metabolites was observed as small round holes appearing, first in the asymmetric apical plasma membrane of terminally differentiated superficial cells, and later in the symmetric apical plasma membrane of exposed undifferentiated intermediate and basal cells. Exposed cells which remained undamaged, immediately underwent maturation of the symmetric apical plasma membrane. These results indicate that CP metabolites perforate the symmetric and asymmetric membranes of most urothelial cells. PMID- 11944065 TI - The small world in biophysical systems structural properties of glycolysis and the TCA cycle in Escherichia coli. AB - It has been shown that the central metabolic network of Escherichia coli is of the small-world type. In this paper, we present that the metabolic network of glycolysis and TCA cycle as a part of the E. coli metabolism is also a small world network. We found that the hubs of the studied network are consistent with those found in the complete metabolic network. The evolutionary meaning of this finding is discussed. PMID- 11944064 TI - The formation of an inverted hexagonal phase from thylakoid membranes upon heating. AB - Barley thylakoid membranes were studied with FTIR and EPR spectroscopy. Thylakoids were exposed to elevated temperatures in order to induce structural changes. As temperatures increased through physiological to even higher levels, no features changed, but upon heating to above 45 degrees C, the fraction of lipid acyl chain segments with gauche-type vibration increased, accompanied by a sharp drop in the membranous spin probe component. These apparently conflicting observations in fact concur with the formation of an inverted hexagonal (H(II)) phase, supporting its putative role in protecting the photosynthetic machinery in thylakoid membranes against thermally-induced disassembly. PMID- 11944066 TI - Sphericity of the femoral head on anterior-posterior radiographs of dysplastic hips. AB - We analyzed the sphericity of the femoral head of dysplastic hips. Using standard anterior-posterior radiographs of the hips, we assessed the femoral head's deviation from a spherical shape using a computer algorithm and via Severin grading. The method presented could serve as a useful tool to quantify differences in sphericity in cases where it is difficult to grade the hip radiologically. PMID- 11944067 TI - Calculation of small angle scattering intensities from molecular dynamics simulation. AB - A method is presented to efficiently calculate small-angle neutron and X-ray solution scattering intensities from explicit - atom model of macromolecules and the surrounding solvent. PMID- 11944068 TI - Electric current density imaging of tablet dissolution. AB - The Electric current density imaging technique (CDI) was used to monitor the dissolution of and ion migration from tablets of different acids in agar-agar gel. Conventional MRI cannot monitor these processes, since it can only show changes in the size of the tablet during the dissolving process. CDI traces the dissolved ions thanks to changes in conductivity. PMID- 11944069 TI - Renal injuries induced by chronic intoxication with microcystins. AB - The microcystins (MCs) LR and YR are hepatotoxins produced by some species of freshwater cyanobacteria. Only a few reports on the acute nephrotoxicity of MCs have been published to date. Here we investigate the effects on rat kidneys of chronic administration of relatively low doses of MC LR and MC YR. Male Wistar rats were injected every second day for 8 months with MC LR (10 microg/kg i.p., n=5) and MC YR (10 microg/kg i.p., n=5). The control group was treated with vehicle, a mixture containing 0.8 % ethanol and 0.2 % methanol in 0.9 % saline (3.7 ml/kg i.p., n=5). We found that MCs could induce damage to the kidney cortex and medulla. The lesions mainly consisted of damaged and dilated tubuli filled with homogenous eosinophil material. We conclude that long-term exposure to relatively low doses of MCs poses a considerable risk for kidney injury. PMID- 11944070 TI - EPR spectroscopy of common nitric oxide - spin trap complexes. AB - Two commonly used hydrophobic and hydrophilic spin traps for NO, namely Fe2+(DETC)(2)and Fe2+(MGD)(2), respectively, were analyzed via EPR spectroscopy. EPR spectra of trapped NO, together with field position standards, were recorded both in the frozen state and at room temperature. We present a detailed characterization of the EPR spectra of the above paramagnetic NO complexes, concerning g-value, hyperfine splitting and linewidths. This study also provides spectroscopic data required to develop a quantitative and sensitive detection system for nitric oxide both in hydrophobic and hydrophilic aqueous media. PMID- 11944071 TI - How should dysplastic human hips be evaluated? AB - Dysplasia of the human hip is characterised by insufficient anterolateral covering of femoral head by the acetabulum. In our study, we evaluated dysplastic human hip joints using biomechanical parameters (the peak contact stress in the weight-bearing area of the hip - pmax) and X-ray image parameters (the centre edge angle - thetaCE, the transverse acetabular inclination angle - thetaUS, the acetabular index of the weight-bearing zone - thetaAC, the ACM angle - thetaACM, and the hip value - HV). The purpose of this study is to make use of X-ray and biomechanical parameters to evaluate hips diagnosed with "hip dysplasia", and to establish whether or not there is a correlation between the two. Our results show a statistically significant correlation between pmax and thetaCE, thetaUS and thetaAC. The correlation between pmax and thetaACM and HV is not statistically significant. PMID- 11944072 TI - The split integration symplectic method. PMID- 11944073 TI - The relationship between innervation density and dynamic balance function. AB - Functional reach (FR) significantly increases with additional sensory information from the fifth metacarpal surface of the dominant hand. The purpose of this study was to determine the relationship between the innervation density of the skin above the fifth metacarpal surface of the palm and the increase of FR with an additional light touch. The results show a moderate negative correlation between innervation density and the increase of FR with light touch. PMID- 11944074 TI - The influence of amino acid side chains on water binding to the copper(II) in bis(N,N-dimethyl-L-alpha-isoleucinato)-copper(II): an EPR and molecular mechanics study. AB - Simulations were done of the electron paramagnetic resonance (EPR) spectra for bis(N,N-dimethyl-L-alpha-isoleucinato)copper(II) dissolved in deuterated methanol as a function of temperature. They indicated different behaviour of the complex below and above 300 degrees K. The effect was examined by the conformational analysis of the copper(II) complex with a new molecular mechanics force field. PMID- 11944075 TI - Quantification of the MR images of hydrogels by NMR relaxation measurements. AB - Layers of hydrogel represent a diffusional barrier that retards the process of drug release. For better prediction of drug release, a method for evaluating the polymer concentration profile was developed. PMID- 11944076 TI - Shape of vesicles in flow--theoretical analysis. AB - The shapes of phospholipid vesicles in a liquid flow with a low Reynolds number were calculated within the nearly spherical approximation. The effective tension model was used in the case of large tension values. PMID- 11944077 TI - Heart rate variability--a shape analysis of Lorenz plots. AB - A new quantitative geometrical method to analyse heart rate variability is presented. The standard image analysis techniques were applied to the grey scale images of Lorenz plots, and the outlines of the attractor areas were determined via a contour following procedure based on a maze walking algorithm. For graphs of simple compact shape, the attractor region contours were described in terms of Fourier coefficients. PMID- 11944078 TI - Selective radiography of 10B distribution in organs using cold and thermal neutron beams. AB - The investigation of boron biodistribution by neutron induced autoradiography was performed using a CR-39 etched track detector. Calibration samples, made of boronated chicken liver, and freeze-dried mouse tissue samples were irradiated both with thermal and cold neutrons. Digital images were made on the basis of track densities, with the selection of tracks due to 10B(n, alpha)7Li reaction. A comparison of results obtained both with thermal and cold neutrons demonstrated that it is possible to obtain equivalent results with both neutron sources, although the background noise due to recoil protons from the fast neutrons of the thermal neutron source is 15 % higher than that of the cold neutron source. 10B concentrations in the range of 0.5 ppm to 150 ppm were determined, with a standard deviation of 13 % and 8 % for the mouse tissue samples and calibration samples, respectively. PMID- 11944079 TI - The influence of Na+ ions on histamine secretion from mast cells induced by NGF or compound 48/80. AB - It has been demonstrated that the release of histamine from mast cells by cytokines is strongly dependent on extracellular Ca2+ and Na+ ions. The results of our current research indicate that the removal of extracellular Na+ enhances NGF induced histamine release, but reduces induction by compound 48/80, suggesting that different mechanisms are involved in the secretory process induced by these agents. PMID- 11944080 TI - Electron paramagnetic resonance characterization of the exopolysaccharide layer produced by bacteria. AB - In order to establish a method to characterize the structural heterogeneity of the bacterial surface, research was conducted with a combination of experiments based on electron paramagnetic resonance (EPR) concentration-imaging (CI) and the modeling of translational diffusion with local restrictions. The benefits and drawbacks of this approach are discussed for the Vibrio sp. exopolysaccharide (EPS) layer. PMID- 11944081 TI - The effect of proprioceptive training on neuromuscular function in patients with patellar pain. AB - Eight patients with patellar pain underwent knee proprioceptive training. The maximal knee extension torque associated with the Vastus Lateralis EMG signal increased (p 0.001 and 0.039). Although muscle balance was not improved, all the patients improved their clinical scores. PMID- 11944082 TI - Alternating electric fields stimulate ATP synthesis in Escherichia coli. AB - External alternating electric fields of low intensity stimulated membrane bound ATP synthesis in starving Escherichia coli cells with electric field amplitudes of 2.5-50 V/cm and a frequency optimum at 100 Hz. The model of electrocon formational coupling was used to analyze the frequency and amplitude responses of ATP synthesis. Two relaxation frequencies of the system were obtained at 44 Hz and 220 Hz, and an estimate of roughly 12 was obtained as the effective charge displacement for the catalytic cycle of ATP synthesis. PMID- 11944083 TI - Importance of a small N-terminal region in mammalian peptide transporters for substrate affinity and function. AB - The two closely related, proton-coupled, electrogenic mammalian peptide transporters PEPT1 and PEPT2 differ substantially in substrate affinity and mode of function. The intestinal carrier PEPT1 has a lower affinity for most substrates than the isoform PEPT2 that is expressed in kidney, lung, brain and other tissues. A previous analysis of PEPT1-PEPT2 chimeras has suggested that the N-terminal half of the carrier proteins is important for substrate affinity. We constructed and analyzed new PEPT1-PEPT2 chimeras for identifying smaller segments within the N-terminal region of the transporter proteins that contribute to the kinetic properties. The first 59 or 91 amino-acid residues of PEPT1 were used to replace the corresponding region in PEPT2 leading to the chimeras CH3 and CH4, which could be analyzed when expressed in Xenopus laevis oocytes. Substrate affinities of both chimeras for the zwitterionic substrate D-Phe-Ala ranged between those that are characteristic for either PEPT1 or PEPT2, but when charged dipeptide substrates were employed, both chimeras possessed PEPT1-like affinities. The chimera CH3 carrying the N-terminal 59 amino-acid residues of PEPT1 exhibited a PEPT2-like phenotype with respect to pHout-dependency as well as to the current-voltage relationship of inward currents. In the chimera CH4 possessing the 91 amino-terminal residues of PEPT1, a pronounced alteration in the pHout-dependence was observed, with highest transport rates occurring at pH values as low as pH 4.0. Based on this analysis, we propose that the two identified aminoterminal regions in mammalian peptide carriers play an important role in determining the substrate affinity and also other characteristic features of the two transporter subtypes. PMID- 11944084 TI - Na+-independent transporters, LAT-2 and b0,+, exchange L-DOPA with neutral and basic amino acids in two clonal renal cell lines. AB - The present study examined the functional characteristics of L-DOPA transporters in two functionally different clonal subpopulations of opossum kidney (OKLC and OKHC) cells. The uptake of L-DOPA was largely Na+-independent, though in OKHC cells a minor component (approximately 15%) required extracellular Na+. At least two Na+-independent transporters appear to be involved in L-DOPA uptake. One of these transporters has a broad specificity for small and large neutral amino acids, is stimulated by acid pH and inhibited by 2-aminobicyclo(2,2,l)-heptane-2 carboxylic acid (BCH; OKLC, Ki = 291 mM; OKHC, Ki = 380 mM). The other Na+ independent transporter binds neutral and basic amino acids and also recognizes the di-amino acid cystine. [14C]-L-DOPA efflux from OKLC and OKHC cells over 12 min corresponded to a small amount of intracellular [14C]-L-DOPA. L-Leucine, nonlabelled L-DOPA, BCH and L-arginine, stimulated the efflux of [14C]-L-DOPA in a Na+-independent manner. It is suggested that L-DOPA uses at least two major transporters, systems LAT-2 and b0,+. The transport of L-DOPA by LAT-2 corresponds to a Na+-independent transporter with a broad specificity for small and large neutral amino acids, stimulated by acid pH and inhibited by BCH. The transport of L-DOPA by system b0,+ is a Na+-independent transporter for neutral and basic amino acids that also recognizes cystine. LAT-2 was found equally important at the apical and basolateral membranes, whereas system b0,+ had a predominant distribution in apical membranes. PMID- 11944085 TI - Claudin 5 is transiently expressed during the development of the retinal pigment epithelium. AB - During the development of chick retinal pigment epithelium (RPE), the permeability and selectivity of the epithelium's tight junctions are continuously modulated. Overall paracellular permeability decreases, but selectivity increases. Because the claudin family of transmembrane proteins appears to provide the structural basis for selectivity, we examined the expression of claudins as a function of development in chick RPE. Degenerate primers were used with the reverse transcriptase-polymerase chain reaction (RT-PCR) to obtain complete sequences of chick claudins 3 and 5. Northern blotting and semi quantitative RT-PCR demonstrated that claudin 5 was expressed in RPE, but claudin 3 was expressed only in the choroid layer of the eye. Northern blotting, semiquantitative RT-PCR and immunoblotting demonstrated that the expression of claudin 5 was transient, with peak levels of expression between embryonic days 10 and 14. Primary cultures were used to demonstrate that factors secreted by the neural retina induced the expression of claudin 5 nearly 3-fold if RPE was isolated from embryonic day 7 embryos. There was little effect if RPE was isolated from embryonic day 14. The upregulation of claudin 5 correlates with permeability changes that occur during the intermediate stage of RPE development. Interestingly, claudin 5 must be replaced during the late stage of development when the number and complexity of tight junctional strands increases. This would imply more changes in selectivity. PMID- 11944086 TI - The oxidant thimerosal modulates gating behavior of KCNQ1 by interaction with the channel outer shell. AB - Thimerosal (o-Ethylmercurithio)benzoic acid, TMS), a membrane-impermeable, sulfhydryl-oxidizing agent, has been described to increase the K+ current IKs in KCNE1-injected Xenopus laevis oocytes. Since there are no cysteine residues in the extracellular domain of KCNE1, it has been proposed that TMS interacts with its partner protein KCNQ1. The aim of this study was therefore to investigate the interaction of TMS with KCNQ1 and the respective K+current IK. In CHO cells stably transfected with KCNQ1/KCNE1, TMS increased IKs, whereas in CHO cells expressing KCNQ1 alone, TMS initially decreased IK. TMS also affected the cytosolic pH (pHi) and the cytosolic Ca2+ activity ([Ca2+]i) in these cells. TMS slowly decreased pHi. With a short delay, TMS increased [Ca2+]i by store depletion and capacitative influx. The time course of the effects of TMS on pHi and [Ca2+]i did not correlate with the effect of TMS on IK. We therefore anticipated a different mode of action by TMS and investigated the influence of TMS on cysteine residues of KCNQ1. For this purpose, KCNQ1wt and two mutants lacking a cysteine residue in the S6 or the S3 segment (KCNQ1C331A and KCNQ1C214A, respectively) were expressed in Xenopus laevis oocytes. A sustained current decrease was observed in KCNQ1wt and KCNQ1C331A, but not in KCNQ1C214A injected oocytes. The analysis of tail currents, I/V curves and activation kinetics revealed a complex effect of TMS on the gating of KCNQ1wt and KCNQ1C331A. In another series we investigated the effect of TMS on IKs. TMS increased IKs of KCNQ1C214A/KCNE1-injected oocytes significantly less than IKs in KCNQ1wt/KCNE1- or KCNQ1C331A/KCNE1-injected cells. These results suggest that thimerosal interacts with the cysteine residue C214 in the S3 segment of KCNQ1, leading to a change of its gating properties. Our results support the idea that not only the inner shell, but also the outer shell of the channel is important for the gating behavior of voltage dependent K+ channels. PMID- 11944092 TI - Considerations for use of the Hoffmann reflex in exercise studies. AB - There continues to be great interest in evaluating the adaptive plasticity of the human nervous system in response to exercise training or other interventions. For various reasons, researchers have been interested in estimates of spinal reflex processing in intact human subjects before and after training. A reflex pathway that has been employed in this regard is the Hoffmann (H) reflex. This brief review describes the basic procedure for evoking the H reflex in different muscles. Other sections address methodological issues that affect interpretation of the H reflex. In particular, the role that presynaptic inhibition serves in the modification of the H reflex and how this precludes its use as an unambiguous measure of alpha-motoneuron excitability will be discussed. Applications of the H reflex to study adaptive plasticity in humans is also reviewed, and methodological requirements that should be maintained for accurate interpretation of H reflexes in exercise studies are presented. PMID- 11944093 TI - Standardising surface electromyogram recordings for assessment of activity and fatigue in the human upper trapezius muscle. AB - The objectives of this work were to determine optimal surface electromyogram (EMG) electrode locations, and inter-electrode distance (IED), when assessing activity and fatigue in the human upper trapezius muscle. Surface EMG signals were recorded from the upper trapezius muscle of 11 healthy male subjects using a linear array of 16 surface electrodes. Five arm positions were investigated (arms at the side of the body, 45 degrees and 90 degrees flexion, 45 degrees and 90 degrees abduction). Fatiguing (1 kg hand load held for 3 min) and non-fatiguing (no load, 0.5 kg and 1 kg hand load held for 3 s) contractions were made. The variabilities of the average rectified value, root mean square, mean and median power spectral frequency and slope over time of these parameters as functions of electrode location and IED (from 5 mm over a range of 35 mm in steps of 5 mm) were quantitatively evaluated. A criterion for selecting the optimal electrode position was applied. This criterion indicated an optimal location measured from the acromion (38% of the distance from the lateral edge of acromion to the spine of the seventh cervical vertebra) which was statistically the same for all the EMG descriptors, arm positions and IED investigated. Finally, it was found that both EMG variables and indexes of muscle fatigue depended on IED which should thus be properly standardised. On the basis of the sensitivity of the EMG descriptors to electrode location and cross-talk reduction, an IED of 20 mm is suggested when a global analysis of activity in the upper trapezius muscle is made using a single pair of electrodes. This study emphasises that a surface EMG analysis of the upper trapezius muscle, following a proper placement of the electrodes and selection of IED, can give reliable indications of muscle activity and fatigue. Data on the myoelectric manifestations of muscle fatigue of the upper trapezius muscle are provided for the optimal electrode location. PMID- 11944087 TI - Functional role of the carboxyl terminal domain of human connexin 50 in gap junctional channels. AB - Gap junction channels formed by connexin 50 (Cx50) are critical for maintenance of lens transparency. Because the C-terminus of Cx50 can be cleaved post translationally, we hypothesized that channels formed by the truncated Cx50 exhibit altered properties or regulation. We used the dual whole-cell patch-clamp technique to investigate the macroscopic and single-channel properties of gap junctional channels formed by wild-type human Cx50 and a truncation mutant (Cx50A294stop) after transfection of N2A cells. Our results show that wild-type Cx50 formed functional gap junctional channels. The macroscopic Gjss-Vj relationship was well described by a Boltzmann equation with A of 0.10, V0 of 43.8 mV and Gjmin of 0.23. The single-channel conductance was 212 +/- 5 pS. Multiple long-lasting substates were observed with conductances ranging between 31 and 80 pS. Wild-type Cx50 gap junctional channels were reversibly blocked when pHi was reduced to 6.3. Truncating the C-terminus at amino acid 294 caused a loss of pHi sensitivity, but there were no significant changes in single-channel current amplitude or Gjss-Vj relationship. These results suggest that the C terminus of human Cx50 is involved in pHi sensitivity, but has little influence over single-channel conductance, voltage dependence, or gating kinetics. PMID- 11944094 TI - The effect of ageing in spinal cord injured humans on the blood pressure and heart rate responses during fatiguing isometric exercise. AB - Groups of 50 healthy male controls and 50 subjects suffering from paraplegia (aged 20-65 years) were examined as to the inter-relationships between age, paraplegia and the strength, endurance, blood pressure and heart rate responses to fatiguing isometric exercise. Contractions were maintained in both groups under voluntary effort and through a contraction induced by electrical stimulation in the paraplegic group. All contractions were maintained to fatigue at a tension of 40% of the maximal muscle strength in either the handgrip or quadriceps muscles. Muscle strength of the handgrip was higher in the paraplegic subjects than in the controls, averaging 589 N and 463 N, respectively for the two groups. In contrast, quadriceps leg extension strength averaged 696 N in the controls and 190 N in the paraplegic groups; for both groups, ageing was associated with a reduction in muscle strength. While leg endurance was less in the paraplegic group than the control group, handgrip endurance was similar in the two groups, endurance increasing with ageing in both the controls and paraplegics. Both systolic and diastolic blood pressures increased at rest in paraplegic and control subjects with age. The magnitude of the pressor response to exercise also increased with age. This was true during both voluntary exercise and exercise induced through electrical stimulation in the paraplegic groups. The heart rate response (change in heart rate during exercise) to a fatiguing isometric handgrip contraction decreased by about 50% between the ages of 20 and 60 years in both the controls and paraplegics for isometric handgrip exercise. In contrast, heart rate changed little with age during contractions of the quadriceps muscle in paraplegics which were induced by electrical stimulation. PMID- 11944095 TI - Pulmonary gas exchange during apnoea in exercising men. AB - There is indirect evidence that cardiovascular responses to apnoea result in a temporary slowing of the O(2) uptake in the lungs in exercising humans. The present study was undertaken in an attempt to determine directly to what extent this occurs, and whether the magnitude of this slowing is such that it must be the result of concomitant cardiovascular readjustments and not merely a result of an isolated apnoea-induced fall in the arterial O(2) saturation ( S(a)O(2)). Eight men performed 120 W leg exercise and performed repeated apnoeas of 10-40 s duration. Heart rate, S(a)O(2), and breath-by-breath gas exchange were determined. Pulmonary O(2) uptake fell gradually as breath-holds proceeded by [mean (SEM)] 74 (3)% of the pre-apnoea O(2) uptake. This decrease was significantly larger than could be accounted for by the fall in S(a)O(2) alone [ S(a)O(2) fall -30 (3)%], which it is estimated would have resulted in a fall of pulmonary O(2) uptake of -54 (5)%. We conclude that cardiovascular responses to apnoea contribute significantly to reducing pulmonary O(2) uptake during apnoea in exercising men. PMID- 11944096 TI - "Occupational backache" - surface electromyography demonstrates the advantage of an ergonomic versus a standard microscope workstation. AB - Musculoskeletal symptoms such as low back pain, neck pain, and tension headache are reported by up to 80% of professional personnel involved in daily microscope work. Yet, in striking contrast to the high prevalence of complaints, there is a general unawareness of this issue both in those suffering, and those ordering and designing microscopes. We intend to call attention to this underestimated work related health hazard and to demonstrate a potential means of prevention. We obtained repeated surface electromyographic (EMG) recordings from the most strained neck, upper limb, and back muscles in 12 healthy volunteers while they were operating a near-to-ergonomic prototype workstation and a conventional microscope, respectively. Mean EMG activity was reduced in all recorded muscles when operating the ergonomic workstation compared to the standard microscope. This improvement became more distinct with sustained work, and was most pronounced in those muscles displaying the highest degree of activity while using the standard microscope. We demonstrate the usefulness of surface EMG recordings to show the advantage of an ergonomically tailored and individually adjustable microscope workstation over a standard microscope. The former allows the operator to maintain a more physiological posture, and may thus prevent the development of cumulative musculoskeletal disorders during prolonged microscope-related work. PMID- 11944097 TI - Maintenance of testosterone status in fitness joggers after increased training mileage. AB - The primary objective was to evaluate the early effect of increased training mileage on testosterone (T) status in recreational joggers. Serum total (T(tot)) and free (T(free)) concentrations at rest, overnight urinary T(tot) excretion, and the T(tot) and T(free) responses to maximal exercise were used as indicators of T status. A group of 13 male [mean (SD) age 24.5 (2.5) years] fitness joggers [maximal oxygen consumption, VO(2max), 52.9 (4.9) ml.kg(-1).min(-1)] qualified as subjects. The training intervention consisted of a 100% increase in the habitual distance run [12 (3) miles.week(-1)] for 2 consecutive weeks, while maintaining the customary training intensity. Blood samples were obtained at rest and after maximal exercise tests, at the beginning and end of a control week of habitual jogging (baseline) and also following the 1st and 2nd weeks of the intervention. The and treadmill exercise endurance time were unchanged across sampling times. Serum T(tot) and T(free) concentrations averaged 565 (62) and 24 (2.6) ng.dl(-1), respectively, at baseline and did not change significantly. Urinary T(tot) excretion averaged 1.5 (0.21) ng.min(-1) at baseline, and also remained unchanged during the intervention. Relative increases in T(tot) (23%) and T(free) (22%) were observed following maximal exercise compared to rest ( P<0.05). However, the exercise-related increases in serum T(tot) and T(free) were not evident after adjustment for the change in plasma volume. It was concluded, that the training intervention did not alter T status in these fitness joggers. PMID- 11944098 TI - Physiological and biomechanical analysis of treadmill walking up various gradients in men and women. AB - This investigation was undertaken to examine the physiological and biomechanical responses during treadmill walking up various gradients in men and women. Groups of 11 men and 11 women took part in four experiments consisting of 8 min treadmill walks at 1.3 m.(-1) up gradients of 0%, 5%, 10% and 15%. The subjects also underwent an anthropometric assessment during which their height, body mass (BM), percentage (%) body fat, lean body mass (LBM), and waist-to-hip ratio (WHR) were obtained. During exercise, each subject was videotaped during three walking cycles so that sagittal kinematics including stride length (SL) and frequency (SF) and ranges of motion achieved at the shoulder, hip and knee during a walking cycle could be determined using a motion analysis system. The motion capture was followed by a measurement of steady-state oxygen uptake (VO(2)) and heart rate (HR) using indirect calorimetry and a HR monitor. Compared to men, women were higher in % body fat, but lower in height, BM, LBM, and WHR. The VO(2) in millilitres per kilogram per minute and HR were similar between men and women at 0% and 5%, but greater in women than men at 10% and 15%. The SL and SF were greater and smaller, respectively, in men than women up all gradients. Shoulder extension was greater up gradients of 10% and 15% in women than men, whereas no sex difference in shoulder flexion was noted. Hip extension and flexion as well as knee flexion remained the same between men and women up all gradients. An inverse relationship was observed between VO(2) in millilitres per kilogram per minute and BM, LBM, and WHR when men and women were combined. In conclusion, the mass-specific metabolic cost of treadmill walking up high gradients is greater in women than men. Such an increase in relative metabolic cost may be attributable to the fact that women are smaller in size, and/or have relatively more BM distributed peripherally. This sex difference may also be due to a greater movement of the upper limbs in women during walking uphill, although direct evidence is needed to support this contention. PMID- 11944099 TI - Norms and correlates of bioimpedance phase angle in healthy human subjects, hospitalized patients, and patients with liver cirrhosis. AB - This study investigates whether bioimpedance indexes rather than derived body compartments would be adequate for nutritional assessment. Evidence is provided that the phase angle as determined by conventional tetrapolar whole body bioelectrical impedance analysis at 50 kHz (1) was largely determined by the arms and legs and not the trunk, (2) was higher in control subjects than in hospitalized patients [mean (SD) 6.6 degrees (0.6) degrees vs 4.9 degrees (1.2) degrees, P<0.001], (3) discriminated poorly between cirrhotic patients of different Child-Pugh class, and (4) was positively correlated with muscle mass ( r=0.53) and muscle strength ( r=0.53) in these patients (each P<0.01). In a prospective study of patients with liver cirrhosis Kaplan-Meier and log rank analyses of survival curves demonstrated that patients with phase angles equal to or less than 5.4 degrees had shorter survival times than patients with higher phase angles [6.6 degrees (1.4) degrees ] and that phase angles less than 4.4 degrees were associated with even shorter survival times ( P<0.01). The prognostic roles of the phase angle and standard nutritional parameters such as total body potassium, anthropometric measurements, and impedance derived fat free mass, body cell mass and fat mass were evaluated separately by Cox regression which eliminated all variables except the phase angle as predictors of patient survival time ( P<0.01). We concluded that for the clinical assessment of patients the phase angle may be superior to commonly used body composition information. PMID- 11944100 TI - An improved Peronnet-Thibault mathematical model of human running performance. AB - Using an improved Peronnet-Thibault model to analyse the maximal power available during exercise, it was found that a 3rd-order relaxation process for the decreasing dynamics of aerobic power can describe accurately the data available for world track records and aerobic-to-total energy ratio (ATER). It was estimated that the time-scales for the decreasing dynamics are around 25 s for anaerobic power output and that they range from 2.12 h to 7.8 days for aerobic power output. In agreement with experimental evidence, the ATER showed a rapid increase during the first 300 s of exercise duration, to achieve an asymptote close to 100% after 1,000 s. In addition, the transition time when the ATER rose above 50% was found to be at a race duration of about 100 s, which would correspond to race distances of about 800 m. The results suggest that the aerobic power output achieves its maximal value at 300-400 s, and reaches a plateau at 26 28 W.kg(-1) that lasts about 5,000 s. After this period, the aerobic power output decreases slowly due to the contribution of long time-scale metabolic processes having smaller energy contributions (about 30% to 40% of the total aerobic power output). PMID- 11944101 TI - The effects of pre-warming on the metabolic and thermoregulatory responses to prolonged submaximal exercise in moderate ambient temperatures. AB - To determine the effects of pre-warming on the human metabolic and thermoregulatory responses to prolonged steady-rate exercise in moderate ambient temperatures and relative humidities [means (SD) 21.7 (2.1) degrees C and 36.7 (5.4)%, respectively], six healthy men each ran at a steady-rate (70% maximal oxygen uptake) on a treadmill until exhausted after being actively pre-warmed (AH), passively pre-warmed (PH), and rested (Cont). Exercise time to exhaustion was significantly reduced following both AH and PH compared to Cont [AH 47.8 (14.0) min, PH 39.6 (16.0) min, Cont 62.0 (8.8) min; P<0.05]. During exercise there were no significant differences in oxygen uptake, total sweat loss, mean skin temperature (T(sk)) and the thermal gradient ( T(re)-T(sk), where T(re) is rectal temperature) following the three conditions. Serum prolactin, plasma catecholamine and plasma free fatty acid concentrations were also similar between all three trials. In contrast, T(re), mean body temperature, heart rate and ratings of perceived exertion were significantly greater during the initial 25 min of exercise following both AH and PH, compared with Cont ( P<0.05). At exhaustion, there were no significant differences in the metabolic and thermoregulatory responses to exercise between the trials. The current findings demonstrate that AH and PH promote a reduction in prolonged submaximal endurance performance under moderate environmental temperatures compared with pre-exercise rest. Such observations appear likely to have been mediated through mechanisms associated with the earlier development of high internal body temperature which resulted in changes in the capacity for heat storage. PMID- 11944102 TI - EMG power spectrum and features of the superimposed M-wave during voluntary eccentric and concentric actions at different activation levels. AB - Muscle fiber conduction velocity (CV) may be affected by the level of voluntary activation and by the diameter of the fiber. Both the frequency component of the electromyography (EMG) power spectrum, such the as median frequency (MF) or mean power frequency, and the duration of muscle compound action potential to single supramaximal electrical impulse (maximal M-wave) may be related to CV. The aim of the present study was to examine how changes in the activation level in lengthening and in shortening conditions would affect the EMG power spectrum during voluntary effort, and compare them to changes in M-wave shape in similar conditions. Ten male subjects performed eccentric and concentric knee extensions at force levels of 40%, 60%, 80% and 100% of maximal eccentric and concentric knee extension force (maximum voluntary contraction, MVC) at an angular velocity of 2 rad.s(-1). In order to measure the M-wave at each force level and in a relaxed condition, a supramaximal electrical stimulus was given to the femoral nerve. The surface EMG was recorded from the vastus lateralis, vastus medialis, and rectus femoris muscles, and the average EMG (aEMG) and MF were calculated. The results show that although the absolute force was greater, the aEMG was generally lower in eccentric as compared to concentric actions at all of the force levels tested. Although the aEMG increased as force increased, no consistent differences were observed in the amplitude of the maximal M-wave in any of the conditions, or in the duration of the M-wave between eccentric and concentric actions. However, as the force level increased the duration of the M wave decreased significantly ( P<0.01) for both eccentric and concentric actions. On average, no major differences were observed in MF between eccentric and concentric actions or between the force levels in either type of contraction, although rather large variations were observed throughout the motions. In voluntary situations, the recruitment of fast motor units with higher muscle fiber CVs and the increased firing rate of the active units increases the muscle fiber CV as the activation level increases. Even though in conditions of supramaximal electrical nerve stimulation all motor units should be activated simultaneously, the duration of the M-wave in the present study decreased as the force level increased. Possible candidates for the change in the CV may be increased activation of the Na(+)/K(+) pump resulting from the activity in the muscle preceding the electrical stimulation and/or changes in the muscle fiber length between different force levels. PMID- 11944103 TI - EMG activity and voluntary activation during knee-extensor concentric torque generation. AB - This study was designed to re-examine and compare the neural drive of the knee extensors during isokinetic concentric muscular actions by means of the twitch interpolation technique (activation level, AL) and surface electromyographic (EMG) recordings (root mean square, RMS). Torque, AL and RMS amplitudes of three knee extensors and one knee flexor were measured in nine subjects during maximal and sub-maximal voluntary contractions, performed under concentric (60 degrees.s( 1) and 120 degrees.s(-1); Con60 and Con120, respectively) and isometric (Iso) conditions. Mean (SD) maximal voluntary torque was significantly lower ( P<0.01) during concentric contractions [Con60: 208.6 (26.8) Nm and Con120: 184.7 (26.4) Nm] compared with isometric contractions [327.4 (52.0) Nm]. A significantly lower AL ( P<0.05) was recorded during Con60 [80.9 (8.8)%] compared with Iso [87.9 (5.1)%] and Con120 [88.2 (6.6)%] maximal contractions. Simultaneously, a lower knee extensor average RMS amplitudes (av.RMS) was measured during Con60 maximal contractions compared with Iso and Con120 maximal contractions. The antagonist biceps femoris RMS values were not different between maximal Iso, Con60 and Con120 contractions. During sub-maximal voluntary contractions, the RMS/torque relationships were similar whatever the muscle considered (vastus lateralis, vastus medialis or rectus femoris) and the AL/av.RMS relationships did not reveal any noticeable differences between each contractile condition. The results of the present study indicate that av.RMS and AL describe similarly the neural drive during maximal and sub-maximal efforts and indicate that during maximal voluntary efforts, neural drive is dependent upon concentric angular velocity (up to 120 degrees.s(-1)). Thus, our results suggest that when applying different contractile conditions, the torque output is regulated via complex interactions between intrinsic muscular properties and the neural drive. PMID- 11944105 TI - [The spiritual dimension. From holy food habits, fasting and starvation to eating without limits measure]. PMID- 11944106 TI - [Celiac disease in adults. The big transformation artist]. PMID- 11944107 TI - [Nutrition counseling. "A small salad and a milk shake" (interview by Urs Luthi)]. PMID- 11944108 TI - [Responsibility in nursing care. More autonomy in routine nursing care]. PMID- 11944109 TI - [Palliative nursing care. "One must do what is possible in order to touch the impossible"]. PMID- 11944110 TI - [The new heart valve]. PMID- 11944111 TI - [Freedom restricting measures--a dilemma for nurses]. PMID- 11944112 TI - [Brief typology of smiling]. PMID- 11944113 TI - [Role of the transplantation coordinator. Between 2 lives: a permanent challenge]. PMID- 11944114 TI - [The smile, an unexceptional value]. PMID- 11944115 TI - [You are what you eat]. PMID- 11944116 TI - [Smiling and psychiatry: must one choose?]. PMID- 11944117 TI - [Auxiliary health personnel for asylum seekers. An original and humanistic definition]. PMID- 11944118 TI - [Social and historical dimension. Nutrition as a mirror of life]. PMID- 11944119 TI - [Personality and human relations. An advantage for nursing practice]. PMID- 11944120 TI - [Professional reintegration in the geriatric milieu. Seizing the chance]. PMID- 11944124 TI - ["It is important to learn from errors"]. PMID- 11944121 TI - [Hepatitis B. Adoption: children at risk]. PMID- 11944125 TI - [Oncology. Recognizing and modifying anxiety]. PMID- 11944126 TI - [Basic stimulation. Stimulating whole body washing]. PMID- 11944127 TI - [Intergenerational relations. Multilevel and ambivalent]. PMID- 11944128 TI - [(Not) a brief awakening]. PMID- 11944129 TI - [Dilemma between autonomy and risk management]. PMID- 11944130 TI - [Growing]. PMID- 11944131 TI - [How to approach helping persons who hear voices. Facing auditory hallucinations]. PMID- 11944132 TI - [Sleep disorders. Would you like something (different) for falling asleep?]. PMID- 11944133 TI - [Bilateral agreement. What changes for frontaliers?]. PMID- 11944134 TI - [Taking leave from the gods in white]. PMID- 11944135 TI - [Questions about death and grief in an EMS (medico-social establishment). Seeing persons die]. PMID- 11944136 TI - [Length of hospitalization. Divergent points of view]. PMID- 11944137 TI - [F as in family]. PMID- 11944138 TI - [Medication safety. An error happened....]. PMID- 11944142 TI - [Patient care network for a better birth]. PMID- 11944143 TI - [Mother and infant protection, what are the stakes for tomorrow?]. PMID- 11944144 TI - [Good birth outcome in Somme]. PMID- 11944145 TI - [Programmed discharge in neonatology]. PMID- 11944146 TI - [Intervention, health education, epidemiology, the role of child health evaluation centers]. PMID- 11944147 TI - ["Did you say 'gavage'?"]. PMID- 11944148 TI - [A room for video EEG in pediatric neurology]. PMID- 11944149 TI - [Nursing care of the newborn infant of the addicted mother]. PMID- 11944150 TI - [Suicidal adolescents, the essential role of school nurses]. PMID- 11944152 TI - [Recommendations for assessing and treatment pain in the child]. PMID- 11944153 TI - [Massage and kinesthetics in the nursing home. And well-being prevails everywhere]. PMID- 11944154 TI - [Denial. Nursing care from the center out]. PMID- 11944155 TI - [Coping by psycho-oncological counseling]. PMID- 11944156 TI - [Rituals in nursing care. At the limit of rational thinking]. PMID- 11944157 TI - [Nursing care and treatment in the final phase of life: a holistic explanation]. PMID- 11944158 TI - [Volunteer work: better than vocation]. PMID- 11944159 TI - [Contact isolation]. PMID- 11944160 TI - [Is shared pain half the pain?]. PMID- 11944161 TI - [Progress can't be stopped. But...]. PMID- 11944163 TI - [Health education of Latin-American women at risk. How enhance access to prevention?]. PMID- 11944162 TI - [Research in medical genetics. The other life of "pedigrees"]]. PMID- 11944164 TI - [Education in oral hygiene for nurses. Basic care may not be neglected]. PMID- 11944165 TI - [Experience of nurses in a drug addiction milieu in Montreal. A "reducing delinquency" program]. PMID- 11944166 TI - [Neuchatel center of physical training. "I also offer them a listening-place" (interview by Francoise Taillens)]. PMID- 11944167 TI - [Share with me your suffering]. PMID- 11944168 TI - [T as in team]. PMID- 11944170 TI - [Well-being also for nurses]. PMID- 11944172 TI - South African nevirapine policy faces growing pressure. PMID- 11944173 TI - New influenza strain and subtype identified. PMID- 11944174 TI - Anthrax update. PMID- 11944175 TI - Indian scientists scouting for new tuberculosis drugs. PMID- 11944176 TI - Retention of variola virus stocks likely to be approved. PMID- 11944177 TI - Spain inaugurates centre for tropical medicine. PMID- 11944178 TI - What makes a Vibrio cholerae pandemic? PMID- 11944180 TI - New drugs needed for tropical diseases. PMID- 11944181 TI - Sepsis definitions. PMID- 11944182 TI - Natural might not be healthier. PMID- 11944183 TI - Saturday night fever: finding and controlling the source of sepsis in critical illness. AB - Fever is a daily concern in the intensive care unit. Although about half of all febrile cases are due to non-infectious causes, fear of sepsis frequently leads to diagnostic tests and escalation of therapy, including broadening antibiotic therapy. Using a case to illustrate this dilemma, we discuss the commonest non infectious and infectious causes of fever, and suggests approaches to their management. Any unexplained fever in intensive care unit patients warrants investigation, which includes complete clinical assessment and blood cultures. When the source of fever is not immediately apparent, non-infectious and infectious causes should be considered. If stable, non-neutropenic patients should be monitored before further tests or empiric antibiotics are started. In an era of rapid emergence and spread of antimicrobial-resistant pathogens and intense scrutiny of resources, optimal diagnosis and management of patients with suspected infection entails much more than the escalation of antimicrobial therapy. PMID- 11944184 TI - Influenza in the acute hospital setting. AB - Influenza poses special hazards inside healthcare facilities and can cause explosive outbreaks of illness. Healthcare workers are at risk of acquiring influenza and thus serve as an important reservoir for patients under their care. Annual influenza immunisation of high-risk persons and their contacts, including healthcare workers, is the primary means of preventing nosocomial influenza. Despite influenza vaccine effectiveness, it is substantially underused by healthcare providers. Influenza can be diagnosed by culturing the virus from respiratory secretions and by rapid antigen detection kits; recognition of a nosocomial outbreak is important in order to employ infection-control efforts. Optimal control of influenza in the acute-care setting should focus upon reducing potential influenza reservoirs in the hospital, including: isolating patients with suspected or documented influenza, sending home healthcare providers or staff who exhibit typical symptoms of influenza, and discouraging persons with febrile respiratory illness from visiting the hospital during a known influenza outbreak in the community. (Note: influenza and other respiratory viruses can cause non-febrile illness but are still transmissible.) The antiviral M2 protein inhibitors (amantadine, rimantadine) and neuraminidase inhibitors (zanamivir, oseltamivir) have proven efficacy in treating and preventing influenza illness; however, their role in the prevention and control of influenza in the acute hospital setting remains to be more fully studied. PMID- 11944185 TI - Superantigens: microbial agents that corrupt immunity. AB - Microbial superantigens are a family of protein exotoxins that share the ability to trigger excessive and aberrant activation of T cells. The best characterised are the staphylococcal enterotoxins and the streptococcal pyrogenic exotoxins that trigger the staphylococcal and streptococcal toxic shock syndromes. It is now apparent that superantigens have a wider role in the pathology of infectious diseases than has previously been appreciated. Staphylococcus aureus and Streptococcus pyogenes together produce 19 different superantigens. The range of microorganisms known to produce superantigens has expanded to include Gram negative bacteria, mycoplasma, and viruses. Research is beginning to shed light on the more subtle parts these molecules play in causing disease and to produce some real possibilities for specific treatment of superantigen-induced toxicity. We aim to highlight these new developments and review the science behind these fascinating molecules. PMID- 11944186 TI - Unlocking the genome of the human typhoid bacillus. AB - Molecular studies are shedding new light on the pathogenesis of human typhoid fever, which is still a very common disease in developing countries. For example, the total genome DNA sequence has recently been determined for a multiple-drug resistant Salmonella typhi, the serotype that is the cause of typhoid fever. The genome sequence showed many distinguishing features, including clusters of S typhi specific genes and a large number--over 200--of pseudogenes. This information, together with other molecular studies, has provided vital clues in several important areas of typhoid biology. We have new insights into the mechanisms underpinning the human host specificity of S typhi, and have exploitable new routes to improved diagnostics and a better understanding of the epidemiology of the disease. PMID- 11944187 TI - Role of lipoteichoic acid in infection and inflammation. AB - Lipoteichoic acid (LTA) is a surface-associated adhesion amphiphile from Gram positive bacteria and regulator of autolytic wall enzymes (muramidases). It is released from the bacterial cells mainly after bacteriolysis induced by lysozyme, cationic peptides from leucocytes, or beta-lactam antibiotics. It binds to target cells either non-specifically, to membrane phospholipids, or specifically, to CD14 and to Toll-like receptors. LTA bound to targets can interact with circulating antibodies and activate the complement cascade to induce a passive immune kill phenomenon. It also triggers the release from neutrophils and macrophages of reactive oxygen and nitrogen species, acid hydrolases, highly cationic proteinases, bactericidal cationic peptides, growth factors, and cytotoxic cytokines, which may act in synergy to amplify cell damage. Thus, LTA shares with endotoxin (lipopolysaccharide) many of its pathogenetic properties. In animal studies, LTA has induced arthritis, nephritis, uveitis, encephalomyelitis, meningeal inflammation, and periodontal lesions, and also triggered cascades resulting in septic shock and multiorgan failure. Binding of LTA to targets can be inhibited by antibodies, phospholipids, and specific antibodies to CD14 and Toll, and in vitro its release can be inhibited by non bacteriolytic antibiotics and by polysulphates such as heparin, which probably interfere with the activation of autolysis. From all this evidence, LTA can be considered a virulence factor that has an important role in infections and in postinfectious sequelae caused by Gram-positive bacteria. The future development of effective antibacteriolitic drugs and multidrug strategies to attenuate LTA induced secretion of proinflammatory agonists is of great importance to combat septic shock and multiorgan failure caused by Gram-positive bacteria. PMID- 11944188 TI - Secrets of success of a human pathogen: molecular evolution of pandemic clones of meticillin-resistant Staphylococcus aureus. AB - The first European isolate of meticillin-resistant Staphylococcus aureus (MRSA) was detected in 1960. Since then MRSA has become a leading cause of nosocomial infections worldwide. Using molecular typing techniques--primarily pulsed-field gel electrophoresis (PFGE)--we identified five major MRSA clones that accounted for almost 70% of the over 3000 MRSA isolates recovered in hospitals mainly in southern and eastern Europe, South America, and the USA. Most of our surveillance studies were done in these areas. Multilocus sequencing typing (MLST) of representative isolates of this collection showed that these five pandemic MRSA clones have evolved from only two distinct ancestral genetic backgrounds, one of which can be traced back to the very first European MRSA isolates and also to meticillin susceptible S aureus strains circulating in Danish hospitals during the mid to late 1950s--i.e., shortly before the introduction of meticillin into therapy. The second lineage with a completely different MLST profile included MRSA frequently recovered in the USA, Japan, and among paediatric isolates from several parts of the world. A few isolates with a third distinct MLST type corresponding to that of EMRSA-16 were also detected in the early Danish isolates. The four structural types of mec element, the heterologous DNA segment containing the meticillin resistance determinant mecA, were present in unique combinations with the MRSA clonal types. Our findings establish evolutionary associations in the most widely spread pandemic clones of MRSA. The epidemiological factors that contributed to the massive dissemination of a few MRSA clones are not well understood. We suggest, however, that the secrets of effectiveness of MRSA could be hidden in the unique genetic background of a surprisingly few lineages of S aureus particularly well able to cope with the contemporary clinical environment. PMID- 11944189 TI - Paula I Watnick--elucidating the role of biofilms. Interview by Pam Das. PMID- 11944190 TI - Creating the position of office safety coordinator. PMID- 11944191 TI - Pouring rights. PMID- 11944192 TI - HIPAA review. PMID- 11944193 TI - Don't underestimate the power of the job description. PMID- 11944194 TI - Effects of a partially hydrolyzed curdlan on serum and hepatic cholesterol concentration, and cecal fermentation in rats. AB - A significant reduction was observed for serum and hepatic cholesterol concentrations in the rats fed diet containing a 5% partially hydrolyzed curdlan (PHCD), whereas only the hepatic cholesterol concentration was decreased in the curdlan (CD)-fed rats. The cecal contents in the CD group contained a significantly larger amount of short-chain fatty acids, but not those in the PHCD group. CD, but not PHCD, significantly increased the population of cecal bifidobacteria. From the in vitro fermentation test with cecal contents from cellulose powder (CP) and CD-fed rats, PHCD proved to be easily fermented by both cecal contents; incidentally CD was more susceptible to the cecal contents from CD-fed rats than to those from CP-fed rats. These results suggest that PHCD is involved in the modulation of lipid metabolism and intestinal microflora through a different manner from the native CD in rats. PMID- 11944195 TI - Cholesterol plus methionine feeding do not induce lipid peroxidation and atherosclerotic changes in the rat aorta. AB - The purpose of this study was to investigate whether cholesterol plus methionine feeding may be a convenient model to produce atherosclerosis in rats, and also to examine the contribution of oxidative stress to this development. For this reason, lipid peroxide levels and antioxidant enzyme activities in the liver and aorta as well as histopathological findings were determined in male Wistar-albino rats fed a diet supplemented with cholesterol plus cholic acid and methionine for six months. This diet was found to increase lipid peroxide levels in the liver of rats. Hepatic glutathione peroxidase (GSH-Px) and catalase (CAT) activities increased, but superoxide dismutase (SOD) activity remained unchanged. In conclusion, cholesterol and methionine feeding in rats did not cause oxidative stress and atherosclerotic changes in the aorta, although hepatic prooxidant antioxidant balance was affected by this diet. PMID- 11944196 TI - Prolonged marginal ascorbic acid deficiency induces oxidative stress in retina of guinea pigs. AB - We examined whether prolonged marginal ascorbic acid deficiency induces oxidative stress in the retina of guinea pigs. Male guinea pigs aged four weeks were given a scorbutic diet (20 g/animal per day) with either marginally deficient ascorbic acid (0.5 mg/animal per day) or adequate ascorbic acid (1 g/animal per day) in drinking water for three and six months. The retinal contents of the reduced form of ascorbic acid in the deficient group at three and six months were 68.1 and 43.5%, respectively, of that in the corresponding adequate group. The retinal contents of the oxidized form of ascorbic acid in the deficient group at three and six months were 1.9- and 2.7-fold, respectively, higher than that in the corresponding adequate group. The content of retinal thiobarbituric acid reactive substances (TBARS), an index of lipid peroxidation, in the deficient group was 2.5-fold higher than that in the adequate group at six months. The retinal contents of reduced glutathione (GSH) in the deficient group at three and six months were 84.8 and 66.7%, respectively, of that in the corresponding adequate group. The deficient group had 37.5% of retinal vitamin E content of the adequate group at six months. The deficient group had higher serum vitamin E concentration than the adequate group in both experimental periods. There were no differences in serum TBARS and GSH concentrations between the groups at both periods. These results indicate that prolonged marginal ascorbic acid deficiency induces oxidative stress in the retina of guinea pigs without systemic oxidative stress. PMID- 11944197 TI - Administration of beta-carotene suppresses lipid peroxidation in tissues and improves the glucose tolerance ability of streptozotocin-induced diabetic rats. AB - The present study attempted to examine the antioxidative effect of dietary beta carotene (BC) on lipid peroxidation (LPO) in the streptozotocin (STZ)-induced diabetic rats. Male Sprague-Dawley (SD) rats were fed on the AIN76 standard diet with or without 0.1% BC. On the 21st day after introduction of these diets, STZ was intraperitoneally injected in half the subjects of both groups. All animals were sacrificed seven days after the STZ injection. Glucose tolerance and thiobarbituric acid reactive substance (TBARS) in the tissues or serum were measured. Body weight gain in the BC + STZ group was significantly higher than that in the STZ group (p < 0.05). Blood glucose and TBARS concentrations of the liver, pancreas, and serum in the BC + STZ group were significantly lower than those in the STZ group. The blood insulin concentration in the BC + STZ group was significantly higher than that in the STZ group. The hepatic and serum beta carotene concentrations in the BC + STZ group were significantly lower than those in the BC group. Moreover, the synthesis and oxidation of glutathione (GSH) in the BC + STZ group were reduced when compared to the STZ group. These results suggest that the administration of beta-carotene suppresses the elevation of LPO and reduces the symptoms of diabetes mellitus (DM) in the STZ-induced diabetic rats. PMID- 11944198 TI - Dose-response effect of docosahexaenoic acid ethyl ester on maze behavior and brain fatty acid composition in adult mice. AB - The dose-response effect of dietary docosahexaenoic acid (DHA, 22:6 n-3) ethyl ester (EE) on maze-learning ability in mice was studied. Male Crj:CD-1 mice aged three months were fed a) a diet containing 5 g palm oil/100 g diet (control group); b) a diet containing 0.5 g DHA ethyl ester/100 g diet plus 4.5 g palm oil/100 g diet (DHA-EE 0.5% group); c) a diet containing 1 g DHA ethyl ester/100 g diet plus 4 g palm oil/100 g diet (DHA-EEE 1% group); d) a diet containing 2 g DHA ethyl ester/100 g diet plus 3 g palm oil/100 g diet (DHA-EE 2% group) for four months. Maze-learning ability was assessed three months after the start of the experiment. The time required to reach the maze exit and the number of times that a mouse strayed into blind alleys in the maze were measured in three trials, performed every four days. In trial 1, the DHA-EE 0.5%, 1% and 2% groups required less (p < 0.05) time to reach the maze exit, and the DHA-EE 2% group strayed (p < 0.05) into blind alleys fewer times than the control group. In trial 3 performed four days after the second trial, the DHA-EE 2% group needed less (p < 0.05) time to find the exit and spent a fewer (p < 0.05) number of times in blind alleys than did the control group. In the total lipids of plasma and brain of mice fed DHA, increasing intakes of DHA resulted in an increase in DHA levels, with a corresponding decrease in arachidonic acid (20:4 n-6). Improved maze-learning ability in mice fed DHA-EE 2% was associated with higher DHA levels in brain. Our resulted suggest that there are no linear dose-response effects of DHA on maze learning ability, however, the intake of DHA-EE 2% diet improves learning ability in adult mice as demonstrated by maze performance. PMID- 11944199 TI - Study on the interaction of ions of transient metals with ascorbic acid in the presence of different scavengers of active oxygen species in SOS chromotest. AB - SOS chromotest was employed to study the interaction of ascorbic acid with free ions of transient metals in the presence of added catalase, superoxide dismutase or D-mannitol. Catalase diminished the genotoxic activity of the mixture of ascorbic acid with copper ions in E. coli strains PQ37 and PQ 300, but genotoxicity of this mixture was not suppressed by superoxide dismutase and D mannitol. The results suggest that copper ions diminished the content of peroxide generated by ascorbic acid. PMID- 11944200 TI - Effects of vitamin C and vitamin E on lipid peroxidation status, serum hormone, metabolite, and mineral concentrations of Japanese quails reared under heat stress (34 degrees C). AB - This study was conducted to determine the effects of dietary vitamin C (L ascorbic acid) and vitamin E (alpha-tocopherol acetate) on lipid peroxidation status measured as MDA and serum triiodothyronine (T3), thyroxine (T4), thyroid stimulating hormone (TSH), adrenocorticotropic hormone (ACTH), as well as some other serum metabolite and mineral concentrations of Japanese quails reared under heat stress (34 degrees C). One hundred-eighty 10-day-old Japanese quails were randomly assigned to six treatment groups, three replicates of 10 birds each. Using a 2 x 3 factorial design, the birds received two levels of vitamin C (100 and 200 mg/kg of diet) or three levels of vitamin E (125, 250, or 500 mg/kg of diet). Greater dietary vitamin E and vitamin C resulted in a greater serum T3, T4, and TSH (p = 0.001), but lower ACTH (p = 0.001) concentrations. Serum concentrations of T4 and TSH increased to a greater extent by increasing dietary vitamin C when greater vitamin E levels were fed (interaction, p = 0.001). Serum glucose, urea, triglycerides, and cholesterol concentrations decreased (p = 0.001), while protein and albumin concentrations increased (p = 0.001) when both dietary vitamin C and vitamin E were increased. Serum activities of SGOT and SGPT were not influenced by dietary vitamin C and vitamin E (p > 0.43). However, serum activity of AP increased (p = 0.001) by increasing both dietary vitamin C and vitamin E. Increasing both dietary vitamin C and vitamin E caused an increase in serum concentrations of Ca, P, K (p = 0.001), Fe, and Zn (p = 0.01) but a decrease in serum concentrations of Na (p = 0.001) and Cu (p = 0.01). Interactions between vitamin C and vitamin E were detected for Ca, P, Na, and K (p = 0.001). Greater dietary vitamin C and vitamin E resulted in a greater serum and liver vitamin E, C, and A (p < or = 0.05), but lower MDA (p = 0.001) concentrations. Results of the present study conclude that supplementing a combination of dietary vitamin C (200 mg) and vitamin E (250-500 mg) offers a good management practice to reduce heat stress-related decreases in performance of Japanese quails. PMID- 11944201 TI - The words private and costly certainly figure large in nurses work. PMID- 11944202 TI - Defining the ethical relationship in clinical care. PMID- 11944203 TI - Interview with Lisbeth Hockey. Interview by Verena Tschudin. PMID- 11944204 TI - Organizational ethics in Finnish intensive care units: staff perceptions. AB - The purpose of this study was to describe ethical problems that are influenced by organizational factors in Finnish intensive care units (ICUs). The goal was to help nurses and administrators to analyse intensive care work, and to improve nurses' work motivation. Through these means the ultimate goal is to improve the quality of patient care. Data were collected in 35 hospital ICUs by means of the Ethics Environmental Questionnaire (EEQ). This gained access to the population of 1047 Finnish intensive care nurses. The response rate was 77% (n = 814). Data analysis was carried out using SAS-6 statistical software. The results provided scores for the 20 EEQ items. Reliability according to Cronbach's alpha was 0.87. The results revealed that organizational factors in Finnish ICUs have both positive and negative dimensions. Positively, nurses have the opportunity to discuss ethical problems in their work units, whereas, negatively, respondents noted that there is much concern about earning money. Nurses' work in Finnish hospital ICUs is ethically challenging; it is similar to that found in other countries and thereby supports international application of these findings. PMID- 11944205 TI - Beyond the boundary between science and values: re-evaluating the moral dimension of the nurse's role in cot death prevention. AB - This article combines a philosophical critique of the idea that public health nurses are primary technicians who neutrally hand over scientifically established facts on risks to the public and an empirical analysis of the actual work of public health nurses. It is argued that the relationship between facts and values in public health is complex and that, despite the introduction of several scientifically-based standards and guidelines, public health nurses are not technicians. They do moral work and experience ethical dilemmas. To get a grip on the specific character of this moral work, we distance ourselves from the idea that there are ethical dilemmas in public health nursing for which we can provide general ethical rules and principles. Instead we suggest a contextual ethical approach, in which several different kinds of consideration may be important. To illustrate this, we analysed 15 in-depth interviews with nurses involved in the prevention of cot deaths in the Netherlands. It is shown that these nurses do not neutrally pass on the epidemiological facts on the risks of prone sleeping, warm bed-clothes and passive smoking, but they are the moral architects of this preventive practice. It is also shown that this moral work and the ethical dilemmas they experience cannot be characterized in terms of general ethical rules and principles. It becomes clear that the moral work of nurses differs according to the three main risks at stake: the balance between virtue, risk taking and responsibility depends on the specific context. PMID- 11944206 TI - Moral problems experienced by nurses when caring for terminally ill people: a literature review. AB - This article is a review of the literature on the subject of how nurses who provide palliative care are affected by ethical issues. Few publications focus directly on the moral experience of palliative care nurses, so the review was expanded to include the moral problems experienced by nurses in the care of the terminally ill patients. The concepts are first defined, and then the moral attitudes of nurses, the threats to their moral integrity, the moral problems that are perceived by nurses, and the emotional consequences of these moral problems are considered in turn. The results show that the moral behaviour of nurses, which is theoretically grounded in commitment to care and to the patient, appears to be shaped by specific processes that lead to engagement or to mental and behavioural disengagement in morally difficult situations. Nurses often appear to fail to recognize the moral dimensions of the problems they experience and also to lack the skills they need to resolve moral problems adequately. Although the findings show that several elements that are beyond the control of nurses, owing to their lack of autonomy and authority, influence their moral experience, intrinsic factors such as feelings of insecurity and powerlessness have a profound effect on nurses' perceptions and attitudes in the face of moral problems. The moral problems perceived by these nurses are related to end-of-life issues, communication with patients, the suffering of patients, and the appropriateness of the medical treatment. PMID- 11944207 TI - Between professional duty and ethical confusion: midwives and selective termination of pregnancy. AB - This qualitative study describes midwives' experiences in relation to termination of pregnancy for fetal abnormalities, and their corresponding professional and ethical position. Thirteen midwives working in a university clinic were interviewed about their problems in this respect. The information gathered was evaluated by using qualitative content analysis. The study focused on the emotional experience of the midwives, their professional position, and ethical conflict. In this situation, midwives are faced with a conflict between the woman's right to self-determination on one hand and the right to life of the child on the other. This conflict causes a high level of emotional stress and, subsequently, professional identity problems. Although questions concerning the child's right to life are generally suppressed, the ethical principle of the woman's right to self-determination is rationalized. Although this process of rationalization seems to present a false ethical decision, it enables midwives to continue with their daily professional duties. As far as orientating midwives to the value of these women's right to self-determination is concerned, it must be assumed that they have made an ethical decision to which they have given insufficient thought. This problem is exacerbated by the fact that midwives are largely excluded from the decision-making process of the parents in question. They cannot therefore help in this process in a valuable and responsible way by providing clear information and proposing objective criteria. In relation to the tasks they are expected to fulfill, these midwives revealed that they were in a state of professional confusion. PMID- 11944208 TI - Nurse autonomy as relational. AB - This article seeks an improved understanding of nurse autonomy by looking at nursing through the lens of what recent feminist scholars have called 'relational' autonomy. A relational understanding of autonomy means a shift away from older views focused on individuals achieving independence, towards a view that seeks meaningful self-direction within a context of interdependency. The main claim made here is that nurse autonomy is, indeed, relational. The article begins with an explanation of the notion of relational autonomy. It then explains both the collective and the individual application of the term 'professional autonomy'. Finally, it argues that both senses of professional autonomy are best understood as relational, and suggests some implications of this conclusion. PMID- 11944210 TI - National Student Nurses' Association, Inc. Code of Academic and Clinical Conduct. PMID- 11944209 TI - An observational study of the level at which parents participate in decisions during their child's hospitalization. AB - When a child is hospitalized, the parents find themselves in an unfamiliar environment and their parental role changes. They are in a stressful and often anxiety-filled situation and it may be difficult for them to participate in decisions. The purpose of this study was to examine the extent to which parents participate in decisions during the course of events when their child is hospitalized. Thirty-five parents of 24 children (aged 5 months to 18 years) were followed by mobile observation during their child's hospitalization in a paediatric department in Sweden. Three researchers analysed field notes in three steps, using manifest and latent coding. In step 1, 119 situations that included a decision process were identified. In step 2, the situations were assessed according to a five-level scale concerning how the parents' wishes, desires or values had been respected. In step 3, each situation was scrutinized with respect to factors influencing the extent of the parents' participation. The results showed that parents have varying ability to be involved in decision making. Professionals need to communicate more openly with them in order to identify and satisfy their needs because some parents are unwilling or incapable of expressing them. PMID- 11944211 TI - [Tuberculosis in Israel--main epidemiological aspects]. AB - BACKGROUND: In the last decade, tuberculosis (TB) has re-emerged as a major worldwide disease and also as a significant disease in Israel, particularly in the context of public health. In 1993, TB was declared a "global emergency" by the World Health Organization and in 1996, it was declared a "dangerous infectious disease" by the Israeli Ministry of Health. AIM: This article aims to provide an epidemiological update on TB to the practicing physician in Israel. We present the perspective of the global situation together with relevant data on TB in Israel for use in clinical decision making. METHODS: All local data are from the ongoing epidemiological surveillance of TB conducted by the Department of Tuberculosis and AIDS in the Israeli Ministry of Health. Other data are cited from international sources. RESULTS: TB is greatly influenced by immigration (some 80-90% of TB cases are foreign-born and at least 65% are among new immigrants). Between 1989-1996 the incidence of TB by cohort analysis for year of immigration ranged from 38-172/100,000 (for new immigrants from the Former Soviet Union) to 500-3,000/100,000 (for new immigrants from Ethiopia). During this period incidence in the veteran population (Israeli-born and immigrants at least 5 years in the country) was, at the most. 4/100,000. Some 80% of TB cases were pulmonary. The rate of drug resistance is increasing some 20% of the isolated strains were resistant to at least one drug and some 8% were resistant to both Isoniazid and Rifampicin. (MDR, multi-drug-resistant). Tuberculosis associated with AIDS has increased in the last decade due to immigration from sub-Saharan Africa. Contrary to the belief existing in the general public, the number of TB cases among foreign workers was relatively low (7.6% of the cases reported in 1998). CONCLUSION: In the clinical context, a practicing physician is most likely to diagnose TB in a recent immigrant. Thus since the symptoms of early TB are non specific, diagnostic efforts (which are labor intensive and expensive) should be guided by a high index of suspicion in that particular population group. Furthermore, this epidemiological data provided the rationale for determining TB control policy as described in the second article on this subject. PMID- 11944212 TI - [Latent tuberculosis infection: diagnosis and treatment]. AB - The essential component of tuberculosis elimination strategy is to identify and treat persons with latent tuberculosis infection (LTBI) who are at high risk for developing active tuberculosis. The tuberculin skin test is the only proven method for identifying LTBI. Although the specificity and the sensitivity are decreased by cross reaction with BCG vaccination and by non tuberculous mycobacteria, there is no better diagnostic tool. The test's positive predictive value is poor in populations with low risk for tuberculosis. Identification of persons with LTBI is focused on groups at high risk who would benefit from therapy (targeted tuberculin testing). The interpretation of the tuberculin skin test reaction is dependent on the risk factors and the immune status of the patient. For the past 30 years, Isoniazid has been the drug of choice for treating patients with LTBI, but its application has been limited by poor compliance and toxicity. Therefore, there has been interest in the development of shorter course treatments such as rifampin (4 months) or rifampin and pyrazinamide (2 months). We describe the new guidelines for targeted tuberculin testing and different treatment regimens for LTBI as recommend by the American Thoracic Society. PMID- 11944213 TI - [Splenic abscess as an unusual manifestation of colonic carcinoma]. AB - A patient was admitted to the Internal Medicine Department after suffering from fever for 4 days. A splenic abscess was found in an ultrasound examination and was confirmed by an abdominal computerized tomography and galium scan. Antibiotic treatment was successful, however barium enema was performed due to the lack of a predisposing factor for this unusual abscess. A polypoid mass was found in the splenic flexure of the colon. Colonoscopy and histologic examination confirmed the mass to be colonic adenocarcinoma. Either an endoscopic examination or barium enema are recommended to exclude colonic lesions in cases of splenic abscess without an obvious cause. PMID- 11944214 TI - [Clinical aspects and treatment of 95 patients with hemifacial spasm]. AB - Hemifacial Spasm (HFS) is considered a peripheral disease of the facial nerve caused by vascular compression at the nerve root in the pontocerebellar angle. We aimed to study the natural course of HFS and especially, to examine the relationships with psychological status or physical activities, in order to assess the possible role of the facial nucleus in the pathogenesis. Ninety-five consecutive patients with HFS, 52 men and 43 women, with a mean age of 62 + 12.7 years and a mean disease duration of 7.5 + 6.5 years, were personally interviewed by 2 of the authors (SB and HK). A detailed questionnaire was completed with direct and indirect questions regarding the relationship between the severity of the spasms and emotional status or physical activity. We found strong association between emotional stress and tiredness and aggravation of the spasms in 85% and 54% of the patients, respectively. Talking increased the spasm severity in 58% of the patients and eating or drinking aggravated the spasm in 28% of the patients. Physical activity, head position, the season of the year or the time during the day had no effect on the clinical status. Botulinum toxin was injected to 78 patients with an overall subjective rate of improvement of > 70% in 74% of the patients (23% graded their rates of improvement as > 90%). In conclusion, HFS is a movement disorder of the facial nerve which is highly influenced by emotional status to support an involvement of the facial nucleus in the pathogenesis. Botulinum toxin is a very effective long term treatment for this disorder. PMID- 11944215 TI - [Fractures of the atrophic mandible--our experience at Tel-Aviv Medical Center]. AB - Fractures of the atrophic edentulous mandible are relatively uncommon. The treatment poses a real challenge due to the underlying medical condition of the geriatric population affected. The complication rate such as infection or malunion is higher compared to fractures in younger patients. Twenty-three cases of fractures of edentulous mandible were reviewed and analyzed. CONCLUSION: The geriatric maxillofacial trauma patients can be treated with open reduction and rigid fixation which allows a quick return to normal function. The conservative approach "close reduction with Gunning-type splints is still an adequate mode of treatment in clinical settings where a more aggressive surgical approach is contraindicated. PMID- 11944216 TI - [Visual field disturbances in epileptic patients treated with vigabatrin (sevril)]. AB - Vigabatrin, one of the newer anti-epileptic drugs (AED), whose effect is mediated via elevated levels of brain GABA, has proved to be effective in drug resistant partial seizures and infantile spasms. Recently, visual field constriction was found in up to 30% of adults, whereas information for the pediatric age group is sparse. We examined 24 visually-asymptomatic children, ages 3.5-18 years, treated for 3.01.6 years at doses of 25-90 mg/kg. These children underwent an ophthalmologic examination, visual evoked potentials, electroretinogram and when possible, perimetry. Over half of the children had at least one abnormal test and 11/17, who were able to undergo perimetric studies, had symmetrical, nasal visual field constriction. In view of the gravity and prevalence of visual field constriction, use of the AED vigabatrin should be weighed against its clinical benefits. PMID- 11944217 TI - [The international day of tuberculosis]. PMID- 11944218 TI - [Facial bone fractures--closed versus open treatment approaches]. PMID- 11944219 TI - [Advances in pathogenesis and molecular biology of endometrial carcinoma]. AB - Cancer research has evolved at a phenomenal pace over the past two decades. Advances in genetics and molecular biology expand the knowledge gathered in the subject. Endometrial carcinoma is a common invasive malignancy in the female genital tract and presents an important diagnostic and treatment challenge. The research aims to develop new molecular tools for early diagnosis and optimal treatment of the disease and its precursors. An updated review of the molecular and genetic research is presented with correlation to the pathological and clinical characteristics of the tumor. PMID- 11944220 TI - [Anti-IgE--a new treatment for allergic diseases]. AB - Immunoglobulin E (IgE) plays a central role in the pathogenesis of allergic diseases. Cross-linking of IgE molecules on mast cell and basophil surface membrane by allergens, triggers the release of multiple mediators, leading to the development of IgE-mediated immediate and late hypersensitivity reactions. In view of the pivotal role of IgE, it became an attractive target for intervention in the treatment of allergic diseases. Murine monoclonal non-anaphylactogenic antibodies directed to the receptor-binding domain of IgE, were found to reduce IgE levels and production. Anti-IgE antibodies have been recently evaluated in several clinical trials involving hundreds of asthmatic patients both adults and children, including patients with allergic rhinitis. Clinical efficacy and good tolerability were demonstrated. This novel therapeutic approach also appears to be promising for potential treatment of patients suffering concomitantly from several allergic disorders. PMID- 11944221 TI - [The new national program for tuberculosis elimination in Israel]. AB - BACKGROUND: The rise in the incidence of TB in Israel, mostly due to immigration from endemic areas, led to the establishment of a new TB control program which follows the guidelines of the World Health Organization (WHO). Reorganization of the TB infrastructure was enabled by specific legislative, administrative and budgetary measures initiated by the Ministry of Health (MOH) in conjunction with the Sick Funds. AIM: To present the theoretical and practical aspects of this new program to the physicians of Israel. PROGRAM OUTLINE: The essence of this program is the use of directly observed therapy (DOT) for all patients together with centralization of TB care in nine national centers, closely supervised by the MOH. This centralization allows a critical mass of patients to be seen in each clinic, thus enabling the labor-intensive task of modern TB treatment, including the supervision and/or the administration of DOT, to be performed in a cost effective manner. Day to day treatment is conducted by general community clinics and supervised by District Health Offices. Hospitalization, a relatively rare necessity in the new program, is available in two dedicated centers with modern isolation facilities. Centralized laboratory services provide timely susceptibility testing. Billing is simplified according to disease categories, using a global assessment of costs negotiated with the Sick Funds. Management and quality control of the program are carried out on an ongoing basis by the Department of TB and AIDS at the MOH. PRELIMINARY OUTCOME: In the first two years that have elapsed since the inception of the program, almost 93% of cases have documented evidence of completion of treatment (under DOT), compared to 54% (without any treatment supervision) before the program came into effect. PMID- 11944222 TI - [The placebo effect--a biochemical basis for a psychosomatic phenomenon]. AB - We review the research over the past decade on the subject of the Placebo Neurochemical mechanism, and it's random presence in the clinic. Our goal is to present the scientific basis of the placebo, and to arouse the awareness of physicians and scientists to the crucial role of placebo in medicine and pharmacological research. The modeling of rationality in modern times displaced the placebo issue to the sidelines of consensus, and it was only around 1950 that scientific interest in the phenomenon reawakened. Today the existence of a Placebo Effect is not doubted--not in everyone, and not at all times, but it's existence is not doubted in relation to groups of people throughout their lives. The involvement of a neurochemical mechanism in the Placebo Effect is also supported by the literature presented, and chemicals such as CCK, Naloxone, Proglumide, and Endorphins seem to effect the neuronal webs mediating the Analgestic Placebo Effect. PMID- 11944223 TI - [Acquired educational deficiency syndrome--AEDS]. AB - In this description of AEDS I would like to present different explanations and diagnostical approaches, preventative measures and therapies for the majority of behavioral disturbances in children and adolescents. In essence, most of these behavioral disturbances in children such as learning disabilities, violence, adolescence problems, appearances of ticks, anorexia, drug abuse and drug addiction are mostly not due to organic reasons, but, in the majority of cases are due to psychological behavioral problems. The roots of these problems are the continuous failure of relationships between the child and the adult who is closest to him/her such as their parents and teachers. Failure stems from the incorrect educational conception of the adults role towards the child and the inability to carry out correct educational theories. Another problem is the inability of the adult to see the interaction between the child and himself through the eyes of the child, and therefore, the adult cannot know and understand what is accepted and processed in the mind of the child. Most adults cannot prevent a natural ingrained reflex to continue supervising over the child even through there is no reasons to do so. Another point to be considered is that over the last few decades children spend more years in their parents home as adults which bring about acute conflicts, creating situations in which one adult is dependent on the other, and has to accept and give in to his ways. Behavioral disturbances do not necessarily come about from extreme oppression such as abuse etc., but from what we call "normal discipline". Oppression may appear on different levels, the extreme being "erasure of ones personality", and of course behavioral disturbances will evolve. Therefore, taking into account the above, it is worthwhile to consider new thoughts and approaches regarding interactions within child-adult relationships. PMID- 11944224 TI - [Hormone replacement therapy and the risk of breast cancer]. AB - There is a growing use of hormone replacement therapy during menopause in the western world. Although most of epidemiologic studies have not found a cause effect relation between this therapy and breast cancer, the fear of breast cancer related to this therapy limits its vast use. Controversy continues to surround the issue of the relationship between postmenopausal hormone therapy and the risk of breast cancer. Physicians, willing to prescribe this treatment, have often been asked by their patients if these hormones can cause breast cancer. Sufficient evidence exists to indicate the possibility of a slightly increased risk of breast cancer with long duration of hormone replacement therapy. However, the epidemiologic data on this cause and effect relation are inconsistent. Therefore, they cannot provide a definitive conclusion on this issue. Furthermore, evidence has shown that women treated with hormone replacement therapy have only localized disease with less metastasis than non-users. This might be due to detection/surveillance bias, but it might also be due to acceleration/deceleration of preexisting disease that reflects our misunderstanding of the breast cancer pathogenesis. Hormone replacement therapy has a cardinal role in the treatment and prevention of immediate and late sequela of menopause such as osteoporosis, cardiovascular diseases, urinary incontinence, etc. A large group of doctors such as family doctors, orthopedics, cardiologist, gynecologists and others recommend this therapy. Patients must also consider all the risks and advantages in their informed decision-making. Therefore we believe that it is important to review this treatment in connection with breast carcinoma. PMID- 11944225 TI - [The effects of hormone replacement therapy (HRT) on the human eye]. AB - HRT (Hormone Replacement Therapy) is the administration of low estrogen preparation to post-menopausal women. This therapy is considered to be effective in protecting various human tissues from aging processes, which are also related to the decrease in endogenous estrogen level. Estrogen receptors were found in many human tissues and organs, including the eye. Therefore HRT may also have a beneficial effect on the eye and its adnexa. This review presents the ever growing data accumulated over the past years concerning the beneficial effects of HRT on various eye pathologies. It was found that HRT may relieve dry eye symptoms in post-menopausal women, may decrease the intraocular pressure and may delay cataract formation in treated women. In the retina, HRT may play a role in decreasing the incidence of Aging Macular Degeneration (AMD) in post-menopausal women. PMID- 11944226 TI - [Subclinical pulmonary dysfunction: an unrecognized diabetes complication?]. AB - The diabetic patient is susceptible to a series of chronic complications and at risk of premature death. The micro- and macrovascular complications of diabetes are very well known. Despite the scarce information on the implications of diabetes for pulmonary function, supporting evidence suggests that the respiratory system is also affected by diabetes. Both collagen and microvascular abnormalities have been proposed as the underlying defects rendering diabetic patients susceptible to respiratory dysfunction. The findings in the few clinical studies that have addressed this question to date are controversial. In this article we review the literature published on this subject. PMID- 11944227 TI - [Pseudotumor cerebri (PTC--an update)]. AB - Pseudotumor cerebri is a condition of intracranial hypertension without localizing signs except for papilledema with normal intracranial contents and normal cerebrospinal fluid constituents. It is seen more frequently in women than in men (8:1) especially women are of childbearing age, and in 90% of cases of obesity. The most common symptoms are headache and visual obscuration. Other symptoms include pulsatile tinnitus, shoulder and arm pain. The papilledema present in almost all PTC patients can lead to decreased vision and blindness. One third of the large series had substantial visual loss including loss of visual field. Treatment has been directed toward preserving vision. Medications that reduce intracranial pressure such as diuretics like Acetazolamide have some success. When vision is threatened, these individuals may undergo optic nerve sheath decompression or lumbar peritoneal shunt to preserve vision. Even with prompt intervention, visual loss can occur. PMID- 11944228 TI - [Transfer of single embryo as a method to reduce twins pregnancy rate in in-vitro fertilization treatment]. AB - INTRODUCTION: The high success rate in IVF treatments is followed by a high rate of multiple pregnancies. Over 30% of IVF pregnancies are multiples and carrying higher risk to the neonates compared with singleton pregnancy. Twin pregnancy is less dangerous then high order multiple pregnancy but it has a significantly higher risk factor compared with singleton pregnancy. Therefore, it is crucial to find methods to reduce twin pregnancy rate. The goal of this review is to present the peril of twin pregnancy and to evaluate the alternative of selective single embryo transfer (SET) in order to reduce the rate of multiple pregnancies in IVF. METHODS: Survey of the literature on SET. RESULTS: Patients with high pregnancy rate bear higher risk for multiple pregnancy. In these cases transfer of single high quality embryo can result in a pregnancy rate of over 30% without the risk of multiple pregnancies. Selective SET may lead to a significant decrease in multiple pregnancies rate with only a slight drop in the general pregnancy rate. CONCLUSIONS: Selective SET in patients with elevated chance to conceive may result in good pregnancy rate without the risk of multiple pregnancy. PMID- 11944229 TI - Market and/or mission? The new impetus for innovation in health care. PMID- 11944230 TI - Reports say insurers most likely to exit markets in face of competition. PMID- 11944231 TI - What patient safety lessons can health care leaders learn from aviation safety? PMID- 11944232 TI - Maternal health negligence claims. PMID- 11944233 TI - [Functional treatment outcome in rectal cancer after trans-abdominal resection with endoprosthesis]. AB - Functional results of treatment of 107 patients with cancer recti, in whom the tumor had localized on the 8-12 cm distance from anus, were analyzed. Comparative estimation of sphincterometric and clinical indexes in terms up to 12 months after performance of abdominoanal resection and transabdominal resection of rectum was conducted. Application of the operation proposed had permitted to quicken and to improve trustworthy the postoperative rehabilitation of patients. PMID- 11944234 TI - [Chemo-irradiation therapy of non-organ malignant tumors of abdominal cavity and retroperitoneal space]. AB - There are presented the results of treatment of 76 patients with nonorganic malignant tumor of abdominal cavity and retroperitoneal space, to whom surgical treatment (radical or palliative one with partial excision of tumor) was not conducted due to various reasons. In 25 patients special treatment was not conducted, in 14--was performed chemotherapy only, in 17--radiation therapy only and in 17--the treatment using radiation plus chemotherapy. Application of special method of treatment have permitted to increase the patients life span, to improve its quality. An advantage of endolymphatic method of chemotherapy in comparison with conventional methods of chemopreparations administration was noted. PMID- 11944235 TI - [Application of hemopoietic cells of embryonal human liver in treatment of pancreatic head cancer complicated by obturating jaundice]. AB - In 30 patients with pancreatic head cancer after surgical treatment of biliary ducts obstructions using biliodigestive shunting method the suspensions, containing stem cells of embryonal liver, were transplanted. The hematological and immunological indexes improvement was noted. After transplantation performance several courses of chemotherapy were conducted, promoting elongation of average life span by 50%. PMID- 11944236 TI - [Iatrogenic esophageal perforation in forcing bougienage of its post-burn cicatricial stricture]. AB - While performing forceful esophageal bougienage for postburn cicatricial stricture its wall perforation have occurred in 9 of 57 patients. All the patients were operated on. For perforation of thoracic esophagus transabdominal transdiaphragmatic mediastinal drainage was performed and for subdiaphragmatic perforation--closure of the defect with Nissen fundoplication, gastrostomy with temporary conduction of ring--like guide fishing-line. One patient died. PMID- 11944237 TI - [Difficulties of diagnosis of pancreatitis and its complications]. AB - Difficulties in diagnosis of an acute pancreatitis and its purulent-necrotic complications are caused by peculiarities of localization of pancreas. Complex estimation of clinical signs, the laboratory and instrumental investigations data are permitting to analyze correctly clinical situation and to choose an optimal variant of the patient treatment program. PMID- 11944238 TI - [Prophylaxis of atrioventricular block in surgery of reentry tachycardia]. AB - Experience of treatment of 85 patients with reentry tachycardia of atrioventricular node was analyzed. In 10 patients (first group) dottal applications directed from coronary sinus ostium toward atrioventricular bundle (AVB) were applied; in 75 (second group)--from the tricuspid valve ring deep inside atrium on the coronary sinus ostium. Temporary disorder of atrioventricular conductivity of different degree was observed in 3 patients of first group and in 1--of the second group, in whom applications were plotted in the middle of distance between coronary sinus ostium and AVB. In one observation several commissures were situated between electrocardiograms of atrium and ventriculus, not permitting to reveal trustworthy the presence of AVB potential. PMID- 11944239 TI - [Laparoscopic intervention in patients affected by Chernobyl AES disaster]. AB - Comparative analysis of laparoscopic interventions was conducted as well as their results and the frequency of complications occurrence in patients, who were the Chernobyl power atomic station disaster survivors and those who were not exposed to radiation. PMID- 11944240 TI - [Recurrence of poly-nodular euthyroid goiter]. AB - Results of surgical treatment of 93 patients with polynodular euthyroid goiter were studied. Influence of the operative intervention volume, morphological form of goiter and postoperative hormonal therapy conduction on the frequency of the recurrency occurrence was noted. PMID- 11944241 TI - [Reflecting the mode of metastasizing of the thyroid gland tumor in TNM classification]. AB - Essential approaches to the regional metastasizing rubric in TNM system were analyzed. Basing on summary of the results of 466 operations, performed for the thyroid gland cancer including intervention on cervical lymph nodes, the subgroup N alteration in international classification TNM was proposed, permitting to reflect individual clinical situation full to the brim and to achieve significant practical gain in further management of patient. The grading proposed corresponds to general principle, according to which every subsequent rubric must determine more severe affection without combining several types of affection in individual patient. PMID- 11944242 TI - [Surgical treatment and comparative characteristics of the general anesthesia course in patients with myasthenia gravis operated on by traditional methods and video-thoracoscopy]. AB - In 1995-2000 yrs 480 patients were operated on in the clinic using videothoracoscopy (VTHS). In 45 of them various forms of myasthenia were diagnosed. In 9 (20%) patients thymectomy was performed. There were no complications. The VTHS application had permitted to shorten the stationary treatment of the patients twice and to reduce the analgetics dosage in postoperative period by 2-2.5 times. PMID- 11944243 TI - [Administration of diprivan during urological surgery in children]. AB - In 56 children, aged from 6 to 14 years, while performing urological operations the narcosis using ketamine was applied and in 43--basing on diprivan and anesthesy with phentanyl. In all the children premedication was conducted using atropine, dimedrol and sibazone. Pain in the vein while diprivan was injected occurred in 6 (14%) of children, involuntary movements--in 16 (37%). While application of diprivan in all the children the mild arterial hypertension was observed, bradycardia was absent on all stages of anesthesy period. The waking up period duration after conduction of anesthesy using ketamine had constituted at average (86.9 +/- 13.1) min, and using diprivan--(35.2 +/- 11.3) min (P < 0.01). Anesthesy basing on diprivan owes significant advantages in comparison with narcosis using ketamine during performance of urological operation in a child. PMID- 11944244 TI - [Criteria of primary multiple bilateral tumors of mammary gland, their clinical significance in diagnosis of primary and metastatic tumors]. AB - Clinical data and results of morphological investigation of the mammarial gland (MG) tumor in 158 women with bilateral affection are summarized. For diagnosis of primary bilateral MG tumors it is possible to use following criterions: absence of remote metastasizing and local recurrency of cancer, spread of the first metachronous or both synchronous tumors no more than of T1-2N0-1M0 stage, presence of nodular form of the MG cancer, localization of tumoral nodes in external hemispheres contralateral MG. In diagnosis of metastatic bilateral tumors may be used criteria, owing acceptable prognostical validity of negative result, presence of remote metastasizing (PVNR 95.2%), spread of the first metachronous or both synchronous tumors of T3-4N0-3M0-1 stage (PVNR 93.6%); infiltrating form of the bilateral MG cancer (PVNR 95.2%); revealing of tumor in contralateral MG during 2 years (PVNR 84.4%); much lower degree of differentiation of the second tumor (PVNR 77.4%). PMID- 11944245 TI - [Application of the apparatus autohemotransfusion in endoprosthesis of big joints]. AB - We observed 16 patients in whom the method of camera intraoperation reinfusion of washed erythrocytes was used during endoprosthetics of joints. For reinfusion we used the system Elmd-500 (Electromedics Inc., USA). The concentration of hemoglobin in peripheral blood remained stable in early and late post-operation period. No complications after transfusion of washed erythrocytes were observed. PMID- 11944246 TI - [Influence of ozone on biochemical and cytochemical indexes of the blood in early period after experimental hepatic trauma]. AB - The influence of isotonic solution of ozonized chloride sodium (OISCHS) on the peroxide oxidation of lipids (POL) intensity, the antioxidant system (AOS) and aminotranspherase enzymes activity, the middle molecular mass peptides (MMMP) concentration in the blood serum and activity of dehydrogenase in neutrophils of peripheral blood in 1, 3, 6, 9 hours after modelling of mechanical dosaged hepatic trauma was studied in experiment on 20 dogs (principal group). OISCHS was infused intravenously by dropper, the ozone concentration was 1.3-1.5 mg/l. Essential bioregulating effect of ozone brought to the system POL/AOS balance normalization, surveying the aminotranspherases and dehydrogenases, activization, stable lowering of MMMP concentration. PMID- 11944247 TI - [Regulation of regeneration processes by synchronous cellular response to the wound cryo-application distance and its influence on wound healing]. AB - Possibility of correcting regulation of regeneration while timely adjusted answer of cells on cryoaffection was established in experiment. In such a situation a small elastic, not protruding cicatrix arises, the healing duration of the wound reduces by 15%. PMID- 11944248 TI - [The guidance of independent work of students during their specialization in pediatric surgery]. PMID- 11944249 TI - [Organizational aspects of the one-staged reconstruction of mammary glands after performance of mastectomy]. AB - The femoral mammarial glands reconstruction after performance of mastectomy have become an ordinary procedure in USA and in countries of Europe. Primary reconstructions owes following advantages: high cosmetic effect, small psychological trauma of female patient, lesser cost of treatment of such patients in comparison with corresponding indices for postponed performance of reconstruction. In Ukraine such operations were not nearly performed yet. An organizational aspects of one-staged reconstruction of mammarial glands using auto tissues after performance of mastectomy for the mammarial glands cancer were presented, basing on literature data and the authors own experience. PMID- 11944250 TI - [Causes of large and complex postoperative hernia of the anterior abdominal wall]. PMID- 11944251 TI - [Methods of trocar wound suture after laparoscopy]. PMID- 11944252 TI - [Diagnosis and treatment of ovarian dermoid cyst in girls]. PMID- 11944253 TI - [Disseminated intravascular blood coagulation syndrome as a response to surgical wound]. PMID- 11944254 TI - [Recurrence of ileus caused by biliary calculus as a complication of choledochoduodenal fistula]. PMID- 11944255 TI - [Paradoxical embolism]. PMID- 11944256 TI - [Diagnosis and treatment of glomus carotid tumor]. PMID- 11944257 TI - [Acute ileus caused by biliary calculus with migration of its fragment in stomach]. PMID- 11944258 TI - [The inborn cyst of common biliary duct]. PMID- 11944259 TI - [Torsion of uterine tube hydatid in a girl simulating acute appendicitis]. PMID- 11944260 TI - [Purulent external cervical pachymeningitis in a child]. PMID- 11944261 TI - [Method of inter-intestinal anastomosis in surgical treatment of complicated colonic cancer]. AB - The elaborated method of colorectal anastomosis formation was applied for the intestinal continuity restoration after its resection for obturative colonic tumoral ileus. Insufficiency of anastomotic sutures was not revealed. Postoperative complications had occurred in 2 (6.9%) of patients. All the patients have survived. PMID- 11944262 TI - [An original method of demolition of re-entry tachycardia of atrioventricular node]. AB - In 75 patients with reentry tachycardia of atrioventricular node the dotted applications were performed on the ishtmus of coronal sinus level, right from the tricuspid valve ring deep to the atrium. The ratio of amplitudes of atrial and ventricular commissures was in limits of 1:10-11:10. After each application the possibility of tachycardia occurrence was tested. It took one procedure to perform to climinate tachycardia in all the patients. The applications quantity had constituted 5.5 +/- 3.8 at average. Roentgenexposition time had constituted- (10.2 +/- 3.8) min. The atrioventricular conduction disorder was not observed. Recurrency of arrhythmia was not noted in 5 months--4 years follow-up. PMID- 11944264 TI - [Strategies of surgical treatment of hemorrhage in postinfection damage of the main vessels in patients with substance dependence]. AB - There was summarized experience of surgical treatment of 88 patients with narcotic dependence, in whom the hemorrhage have occurred due to postinjection injury of the main vessels. Reconstructive-restorational operations were conducted in acute period in the absence of sepsis, hemorrhagic shock and vast focus of infection, using auto vein and in the remote period--synthetic prostheses. All the patients, suffering narcotic dependence, admitted to the clinic, are considered as probably AIDS-infected. Medical staff, performing medicinal care in such patients, must be provided with the individual defense facilities. PMID- 11944263 TI - [Collateral compensation of blood flow and hemodynamics of the lower extremities in atherosclerotic occlusion of abdominal aorta]. AB - Results of investigation of collateral blood flow in 159 patients with atherosclerotic occlusion of abdominal aorta of various kinds were presented. There was shown, that a. mesenterica superior constitutes the main visceral branch, securing the blood circulation compensation in total, high and middle occlusion of abdominal aorta and in low occlusion--a. mesenterica inferior. The change of direction and enhancement of blood flow along a. epigastric inferior was noted in all the patients. In spite of identity of total volumetric blood flow along the collateral branches, the volumetric blood flow velocity along the lower extremities arteries is determined by the level and spread of aortal occlusion. PMID- 11944265 TI - [Prophylaxis of venous thromboembolic complications in patients with hematological diseases after laparoscopic splenectomy]. AB - In 8 patients, to whom laparoscopic splenectomy was performed, the measures for venous thromboembolic complications (VTHEC) prophylaxis were done. The expediency of application of low molecular weight heparin clecsan was substantiated, its advantages over non-fractionized heparin were depicted. Laparoscopic splenectomy permits to minimize the risk of the VTHEC occurrence in hematological practice. PMID- 11944266 TI - [Specifics of sub-fractional serum composition in patients with acute cholecystitis depending on the gallbladder destruction degree]. AB - Subfractional composition of the blood serum proteins was studied up in 120 patients with an acute cholecystitis. Using laser-correlational spectroscopy it is possible to perform differential diagnosis of destructive and nondestructive forms of an acute cholecystitis with 78-80% trustworthiness, what may be applied for express-diagnosis and for conduction of further diagnostic and medicinal measures. PMID- 11944267 TI - [The meaning of interleukin-6 in chronic biliary pancreatitis]. AB - The interleukin-6 (IL-6) contents in the blood serum and in bile from the gallbladder was studied in 25 patients with uncomplicated cholelithic disease and with that, complicated by chronic biliary pancreatitis. Laparoscopic cholecystectomy was done in all the patients. In uncomplicated cholelithic disease IL-6 was not revealed in the blood serum and in majority of observations- in bile from gallbladder. Direct dependence was established between duration of cholelithic disease existence, complicated by chronic biliary pancreatitis, and concentration of IL-6 in bile from gallbladder. PMID- 11944268 TI - [Exudative pleurisy in patients with diabetes mellitus]. AB - Peculiarities of clinical course, diagnosis and results of treatment of exudative pleurisy (EP) were studied in 65 patients with diabetes mellitus. According to the data of complex investigation obtained, using clinico-radiological, laboratory, instrumental methods (transthoracic puncture, thoracoscopy with pleural biopsy), pleural tuberculosis was diagnosed in 60% of patients, nonspecific pleuritis--in 21.5%, pleural mesothelioma--in 10.8%, primary pulmonary cancer--in 5 (7.7%). In 26 (40%) patients pulmonary operation was performed to eliminate the main focus of the EP occurrence. Direct dependence of the course, result of treatment of patients with EP from the type, severity and timely diagnosis of the disease was established. PMID- 11944269 TI - [Clinico-morphological characteristic of the thymus gland changes in myasthenia and their surgical treatment]. AB - Histological investigation of thymic gland was performed in 155 patients with various forms of myasthenia. Two variants of histological changes in thymic gland were revealed. In presence of the first variant performance of thymectomy was effective in 81.3% and in presence of the second--in 51%. PMID- 11944270 TI - [Changes in cell immunity indexes under the influence of thymalin, thyroxine and fibronectin in patients with hyperplastic diseases of thyroid gland before and after the surgery]. AB - Changes in the cell immunity indexes under the influence of thymalin, thyroxin and fibronectin in vitral loading models were studied up in patients with hyperplastic diseases of the thyroid gland. Disbalance of the cell link immunity was revealed before the operation and, in particular, in the main subpopulations of lymphocytes as well as immunoregulating index lowering, which had intensified after the operation. In patients with toxic, nodose euthyroid goiter and chronic autoimmune thyroiditis the existence of (thyroxin- and thymalin) dependent control of the cell link immunity was determined. Application of fibronectin substituting therapy for correction of the hormone depending functional activity of lymphocytes in the early postoperative period in patients with hyperplastic diseases of thyroid gland was pathogenetically substantiated. PMID- 11944271 TI - [Substantiation of application of the anterior trans-pyramidal approach in skull base tumors]. AB - The microtopographic-anatomic substantiation of anterior transpyramid approaches to the skull base tumors was conducted. The transpyramid approaches introduction in clinical practice had permitted to improve surgical treatment results in patients with the difficult of access skull base tumors. Indications and choice of approach depending on the kind, localization and tumor spread in the skull base in the temporal bone pyramid part were substantiated. PMID- 11944272 TI - [Operative treatment of secondary hyperparathyroidism in patients with terminal chronic renal insufficiency in programmed hemodialysis]. AB - The phosphorous-calcic metabolism disorders and secondary hyperparathyroidism (HPTH) occurs nearly in all patients, to whom programmed hemodialysis (PHD) is applied. In majority of observations these disorders become compensated while administration of cholecalciferol (vitamin D3), calcitriol and etc. In 23 patients surgical treatment was performed to correct these disorders. Performance of subtotal parathyroidectomy (STPTHE) had promoted biochemical indexes normalization and the secondary HPTH clinical manifestations regression. The medicamental therapy nonefficacy, progressing osteoporosis, cutaneous itch, which is not connected the PHD procedure, raising of content of the ionized calcium and the alkaline phosphatase activity as well in the blood serum constitute indications for the STPTHE conduction. PMID- 11944273 TI - [Reconstructive microsurgery in the treatment of brachial plexus injury]. AB - Experience of treatment of 157 patients with the plexus brachialis (PB) injury was summarized. The author delineates three clinical groups depending on the kind of the PB autoplasty done. High efficacy of the PB autoplasty with the nerves transplants bed plasty using fascial-subcutaneous flap on vascular peduncle was noted. PMID- 11944274 TI - [Application of eloxatin for the treatment of widespread colorectal cancer]. PMID- 11944275 TI - [The assessment of risk factors of cardiovascular complications in abdominal surgery]. PMID- 11944276 TI - [Application of diprivan (propofol) in newborn babies and in junior age children]. PMID- 11944277 TI - [Changes in immune status in patients with infectious endocarditis in surgical treatment involving artificial blood circulation]. AB - In 98.2% patients with infectious endocarditis, admitted to the hospital for surgical treatment, the moderate immune deficiency state is revealed with the cell link of immunity depression and phagocytic function of neutrophils insufficiency, which become aggravated after operative intervention performance, especially using artificial blood circulation. The cell link of immunity depression, the B-lymphocytes function inhibition are preserved during 6 months after the operation. PMID- 11944278 TI - [The method of performance of the lower extremities amputation on the hip level in vascular diseases]. PMID- 11944279 TI - [The minimally traumatic method of the urinary duct extirpation]. PMID- 11944280 TI - [Universal medico-diagnostic puncture needle]. PMID- 11944281 TI - [Experience in the treatment of children with bezoar]. PMID- 11944282 TI - [Rare observation of malignant mediastinal mesenchymoma]. PMID- 11944283 TI - [Observation of gastric cyst in a child]. PMID- 11944284 TI - [Postoperative hypercholerrhea in a patient with obstructive jaundice]. PMID- 11944285 TI - [The process of establishment and development of cardiological anesthesiology in the Ukraine]. PMID- 11944286 TI - [Hospital infection and rational antibacterial therapy in conduction of operative interventions in cardiac surgery]. AB - The results of bacteriological monitoring in intensive care units of ICVS AMS of Ukraine, specializing on the treatment of patients after conduction of operation on "open heart", were analyzed. In 1998-2001 yrs the S. Epidermidis and Enterobacter strains had constituted the most frequent causes of septic state in patients. Experience of application of different antibacterial preparations was summarized, the main principles of their administration were formulated. Modern approaches to the treatment and prophylaxis of purulent-septic complications were depicted. PMID- 11944287 TI - [Surgical treatment of postop abdominal wall hernia]. AB - Two methods of combined plasty of postoperative abdominal wall hernia (PAWH), based on biochemical conception of pathogenesis of the disease, were proposed. Application of the methods had allowed to close the defect of any size of anterior abdominal wall. For the 1996-2001 period 221 patients with large, huge and giant PAWH were operated. Local and general complications in 29 (13.1%) patients had occurred, one patient died. Result of treatment in term up to 6 years was studied, recurrency of the disease was not revealed. PMID- 11944288 TI - [Diagnosis of obturator canal hernia]. AB - The canalis obturatorius hernia was revealed in 3 women aged 74-84 yrs old of 221 patients who have died due to strangulation of hernia. Diffuse peritonitis, caused by an acute strangulation ileus, was the reason of the death. Osteometric investigations were performed in 50 cadavers. For the improvement of the canalis obturatorius hernia diagnosis there was proposed the application of dots of its projection. PMID- 11944289 TI - [Efficacy of reintervention in inguinal hernia in boys]. AB - Reintervention for inguinal hernia recurrency was performed in 15 boys, constituting 7.4% from general quantity of patients with inguinal hernia. Modified method, elaborated in the clinic of pediatric surgery, was applied. Introduction of medicinal intra- and postoperative complex had promoted the improvement of results of operative treatment for inguinal hernia recurrency. PMID- 11944290 TI - [Experience of surgical treatment of the patients with post-burn cicatricial gastric stenosis]. AB - Experience of surgical treatment of 48 patients with postburn cicatricial gastric stenosis was presented. In 28 of them the combined gastric and esophageal stricture were reveal. Gastric resection according to Billroth-I was performed in 26 patients, gastroduodenostomy according to Finney--in 4, gastric resection according to Hoffmeister-Finsterer--in 5, gastroenterostomy according to Belfler Brown--in 10 and other operations--in 3. One patient died. For the treatment of combined stricture esophageal passability was restored using bougienage, optimally--along the ring-like thread-guide, conducted, while performing gastric operation, through the mouth, esophagus, stomach and brought out via microgastrostomy. PMID- 11944291 TI - [Prophylaxis of gastrointestinal hemorrhage in patients with noninflammatory pancreatic disease complicated by jaundice]. AB - Frequency, severity of clinical course, pathophysiological mechanisms of occurrence and consequences of gastroduodenal erosion and ulcer, complicated by hemorrhage, were analyzed in patients with noninflammatory diseases of pancreas, complicated by jaundice, by cancer, in particular. Basing on investigations done there was elaborated complex programme of prophylaxis of gastrointestinal hemorrhage occurrence in such patients. PMID- 11944292 TI - [Application of synthetic neuropeptides in complex of surgical treatment of acute pancreatitis and its complication]. AB - In experimental pancreatitis conditions there were studied changes in bloody and lymphatic capillaries of pancreas, quantity and functional state of endocrine cells of duodenum under the influence of synthetic neuropeptides dalargin and sandostatin. Results of complex examination and surgical treatment of 126 patients with various forms of an acute pancreatitis were analyzed. The expediency and efficacy of dalargin application in complex of treatment of such patients were proved. PMID- 11944293 TI - [Acute mesenteric ischemia complicated by intestinal necrosis: choice of strategies of treatment]. AB - Treatment of intestinal necrosis, caused by an acute mesenterial ischemia, represents the most complex field of surgery. Results of surgical treatment of 74 patients with various forms of mesenterial ischemia, complicated by intestinal infarction, were analyzed. Revascularization ought to be conducted if the ischemia may be eliminated. High frequency of anastomotic sutures insufficiency is noted in performance of primary resection of intestine. Syndrome of the polyorgan dysfunction constitutes the main cause of death in patients after the operation. PMID- 11944294 TI - [Surgical treatment of integumentary tissues of foot and shin using adhesive dermal tension and cryotherapy]. AB - The first experience of performance of plasty for nonhealing wounds and ulcers of foot and shin, based on grafting of the surrounding skin on the defect region, using stimulating atraumatical adhesive dermotension independently and in combination with cryotherapy, was presented. The methods proposed were applied in 21 patients for restoration of bearing and other tissues of foot and shin, when defect was up to 110 cm2, since 1996. Positive result of treatment was noted. PMID- 11944295 TI - [Surgical strategies in spontaneous rupture of renal allograft]. AB - The methods of surgical treatment of spontaneous rupture of renal allotransplant (RAT) were studied up in 21 patients. In 17 (80.9%) of patients the RAT rupture was caused by an acute reaction of rejection (ARR), in 14.3% of observations--by an acute necrosis of tubules, in 2 (9.5%)--by renal vein thrombosis and in 2 (9.5%)--by an ischemic damage of the transplant. Surgical treatment of the RAT spontaneous rupture (except the cases with rupture due to renal vein thrombosis) must be managed by the hemorrhage stoppage, the RAT tissues strain reduction (decapsulation and the cross-like capsulotomy), conduction of its intraoperative anticrisis and antiischemic defense with subsequent complex therapy for ARR and the transplant dysfunction. Application of such a tactics of treatment have permitted to secure the RAT and its functions in spontaneous rupture in 82.4% of observations. The transplant survival was registered in terms up to one year in 64.7% of observations. PMID- 11944296 TI - [Experience of application of meronem in the treatment of severe purulent sepsis in children]. AB - Results of antibacterial therapy with application of meropenem (meronem by firm Astra Zeneca, Great Britain) in 29 childs with severe purulent-septic state, including 5 with toxic peritonitis, in 2--with sepsis, in 3 newborn childs--with esophageal atresia and in 19--with the blood disease (nonhodgkin's lymphoma, an acute lymphoblastic or myeloblastic leucosis, reccurrent of acute lymphoblastic leucosis, nephroblastoma). PMID- 11944297 TI - [The change of hemodynamics index in acute period of severe combined injury]. AB - Clinical significance and prognostical informativity of the hemodynamics indexes in an acute period of traumatic disease (TD) in severe combined thoracoabdominal trauma (SCTAT) were studied. The cardiac pump function, tone of vessels, their resistance, venous return were estimated. The pronounced and stable disorders of the hemodynamics indexes in injured persons with SCTAT were noted, their degree depended on the trauma severity, which had determined the TD outcome. PMID- 11944298 TI - [Palliative methods of surgical treatment of Ebstein anomaly]. AB - Ebstein's anomaly--is a complex disease of the heart, the main components of which are dysplasia and tricuspid valve malposition, causing hemodynamical disorders. Principal method of correction of this anomaly is surgical one. There are many surgical methods of correction of the anomaly proposed: tricuspid valve prosthesis, plastic operation, palliative methods. Indications for surgical treatment of Ebstein's anomaly is cyanosis, the blood flow insufficiency, severe disorders of the heart rate. PMID- 11944299 TI - [Radical surgical treatment of heart myxoma]. AB - Experience of surgical treatment of 362 patients with the heart myxoma (HM) was summarized. The expediency of application of various approaches in the left auricle (LA) myxoma was analyzed. Significant advantages of application of the approach through the right auricle to reduce the danger of the tumor fragmentation while its excision were noted. There was elaborated and introduced transbiauricle approach to the LA tumor, application of which secures minimal risk of the tumor fragmentation as well as the possibility of adequate heart revision conduction and the processing of full value of tumor bed for any localization. In terms of observation up to 30 years in 5 (17%) of patients the HM recurrency had occurred. Application of the method, which envisages excision of part of interatrial septum, where HM is fixed, in its all thickness with subsequent suturing and plasty of defect using the patch, making additional processing of the tumor bed with iodine solution and diathermocoagulation, had permitted to reduce the recurrency of the disease from 4.3 to 0.5%. PMID- 11944300 TI - [The method of treatment choice for the abdominal wall hernia]. AB - Experience of treatment of 975 patients with postoperative and recurrent abdominal wall hernia (AWH) and of 1473 patients with inguinal hernia was summarized. Intraoperative classification of Shevrel and Rets for AWH and of Gilbert and Rutkov for inguinal hernia were applied. The conventional methods of plasty, using local tissues, were performed up to 1998 in 648 patients with AWH and in 983--with inguinal hernia. Synthetic implants were used since 1999 in 327 patients with AWH and in 181--with inguinal hernia. In application of conventional methods recurrency had occurred after plasty for AWH in 26.6% and for inguinal hernia--in 5.8% of observations; hernial recurrency was not noted after usage of the modern plasty methods. PMID- 11944301 TI - [Remote results of mini-invasive sympathectomy in chronic arterial insufficiency of extremities]. AB - Results of application of miniinvasive desympathization in 42 patients with chronic arterial insufficiency of extremities were analyzed. Clinical and functional dynamic changes of peripheral arterial bed before and after conduction of desympathization in the early, late and remote postoperative period were noted. PMID- 11944302 TI - [Three stage testing control of students' knowledge in general surgery course]. PMID- 11944303 TI - [Methods of suturing in pyloroduodenal perforative ulcer]. PMID- 11944304 TI - [Complications in patients operated on for perforative duodenal ulcer]. PMID- 11944305 TI - [Mediastinoscopy in the diagnosis of surgically treated mediastinal diseases]. PMID- 11944306 TI - [Complex treatment of acute and chronic cholecystitis in central regional hospitals]. PMID- 11944307 TI - [Abdominal tuberculosis]. PMID- 11944308 TI - [Experimental testing of a method of partitioning segregating populations based on component genotypes]. AB - Partitioning method by L. Powers allows one to obtain true Mendelian description of quantitative trait inheritance, which makes possible a selection prognosis. PMID- 11944309 TI - [Chromosomal localization of a gamete-killing gene in a wheat line]. AB - Monosomic analysis revealed that a gametocidal gene in a common wheat line derivative from Aegilops speltoides is located on chromosome 2B. This gene induces semisterility, low level of chromosome breaks, shriveled seeds, and endosperm degeneration. The obtained data indicate that the localized gene is allelic to well known gametocidal genes Gc1a and Gc1b. PMID- 11944310 TI - [Genetic classification of hordeins by PAAG electrophoresis]. AB - New approach to genetic classification of hordeins based on identification of allelic state of hordein-coding loci by means of the PAAG electrophoresis and establishing the genotype formulas is proposed. PMID- 11944312 TI - [Molecular and genetic polymorphism of cultivated soybean detected by AP-PCR, SSRP and SSR]. AB - Molecular-genetic polymorphism of cultivated soybean from different geographic regions was analysed using three PCR methods (AP-PCR, SSRP and ISSR). The dendrogram of phenogenetic relationship was constructed on the base of obtained data. The allocation of varieties are in a good agreement with pedigree information. Main parameters such as polymorphism level, expected heterozygosity, and marker index were calculated. It was shown that ISSR is the method of choice for the differentiation of soybean varieties. PMID- 11944311 TI - [Frequency-dependent sexual selection in a natural population of oak leaf-roller moth (Tortrix viridana L.)]. AB - A mating system on the esterase locus Est-4 was studied in a natural population of oak leaf-roller moth Tortrix viridana L. Frequency-dependent sexual selection was revealed, which manifested itself in a "rare-type advantage". On the basis of obtained data, a new hypothesis for explanation of the "rare-type advantage" phenomenon is proposed. The hypothesis proceeds from the genetically determined choice of sexual partner, depending on the degree of phenotypic likeness. PMID- 11944314 TI - [Role of peptide complexes from the kidney and thymus in the regulation of apoptosis in lymphoid cells]. AB - Using morphological and electrophoresis methods special features of apoptosis induction by peptide complexes from kidney and thymus were analyzed in thymocytes and peripheral blood lymphocytes of pigs. It was shown that peptide complexes both from kidney and thymus induced thymocytes apoptosis in an allogenic system. In these conditions, genetic distinctions between individuals and a degree of thymocyte maturation determined the level of apoptosis. PMID- 11944313 TI - [Allele polymorphism of micro- and minisatellite loci in populations of different regions of Ukraine]. AB - The results of molecular-genetic analysis of 12 mini- and microsatellite loci in populations of different regions of Ukraine (Kiev, Kremenchug, L'vov, Lugansk, and in Crimea tatars) were presented. Allele frequencies for each locus were determined and genetic distances between analyzed populations were calculated. The results of the analysis were applied for investigation of genetic heterogeneity and biological history of populations from different regions of Ukraine. PMID- 11944315 TI - [Ultrastructural and morphometric characteristics of neutrophilic granulocytes of blood]. AB - Ultrastructural and morphometric characteristics of neutrophilic granulocytes (NG) of blood were investigated. It was found that every cell size fraction distinguished by its own morphologic type of NG and its specified quantitative and qualitative characteristic of primary granules. There is a close interrelation between morphometric and ultrastructural characteristics, and such parameters as perimeter, area of NG, and area of primary granules can be considered as criteria of the NG functional state. PMID- 11944316 TI - [Risk factors of complicated postnatal period in low birth weight infants]. AB - Antropometric indices have been studied in 952 newborns. It has been shown those children with middle mass and height indices in birth have the most expressed homeostatic resources. Children with low body mass in birth are characterized by the decreasing of resistance and high morbidity. Phenotype B, incompatibility with mother according to ABO, Rhesus systems are the risk factors of complicated neonatal period course that results from the disturbances in balance of regulatory lymphocyte subpopulations. PMID- 11944317 TI - [Ultrastructure and metabolism of cardiomyocytes under experimental stress]. AB - Morphogenesis of adaptational, disadaptational, and alterative changes affected structure and function of all live-supporting systems (contractile elements, macroergic phosphate synthesis, intracellular regeneration, and electrolyte exchange control) was studied in cardiomyocytes under experimental chronic stress using ultrastructural and cytochemical methods. PMID- 11944318 TI - [Population prevalence of congenital hypertrophic pyloric stenosis in the Donetsk region of Ukraine]. AB - The population prevalence of congenital hypertrophic pyloric stenosis in Donetsk region in the course of 11 years (1989-1999) has been descriptively studied. Its frequency is 0.73:1000 or 1:1370 newborn infants annual with range of 0.54 to 1.01:1000. It is somewhat lower than that of more wide spread congenital diseases: labial cleft and hemochromatoses. Pyloric stenosis is detected in 42 of 43 populations of Donetsk region. There is a population nonuniformity of the case rate: from 0.19 to 1.9:1000 newborn infants, that the authors are inclined to explain by dissimilarity of a gene pool in the populations due to random migration of the population. PMID- 11944319 TI - [Peculiarities of embryonal development in vitro of Brassica napus pollen grains]. AB - Literature data concerning the problem of the embryogenesis in microspore cultures of Brassica were reviewed. The influence of stress factors on the microspore cultures was examined. The some cytological and morphological peculiarity microspore entering in the embryogenesis developmental pathway were described. PMID- 11944321 TI - Genetic variability of somatic embryogenesis in tissue cultures of sugar beet breeding lines. AB - Genetic variability of callus initiation and plant regeneration has been investigated among three sugar beet genotypes. It was found that TDZ has a genotype-independent effect on callus initiation and is responsible for more than a two-fold increase in the friable callus induction rate and more than a three fold increase in the shoot regeneration rate from this callus. Along with the genotype-independent organogenesis, regeneration from callus occasionally went through the process of somatic embryogenesis in a highly genotype-specific manner. Despite fast and uncontrollable conversion of embryos to normal plants, it was possible to select and maintain repetitive embryogenic culture without loosing regeneration and root formation capabilities. Extensive experimenting with medium composition and culture conditions resulted in an optimal medium for maintenance of repetitive embryos. Comparing with BAP, low concentrations of TDZ provide higher level of adventitious shoot formation and do not induce vitrification of tissues. PMID- 11944320 TI - [Conjugation of the 1BL/1RS translocation with qualitative and quantitative traits in the common wheat T.aestivum]. AB - The review considers the effect of the rye 1BL/1RS translocation in the common wheat genome on qualitative and quantitative traits: grain quality, resistance to diseases, productivity and adaptivity, parthenogenesis, regeneration in anther culture, frequency of chromosome aberrations and frequency of cross-pollination. Data on special features of transmission of the 1BL/1RS translocation through male and female gametes are presented. PMID- 11944322 TI - [Differentiation, identification and development of database of T. aestivum L. varieties of Ukrainian selection on the basis of sequence-tagged analysis of microsatellite repeats]. AB - Determination of the variety genotype is very important for the development of theory and practice of plant breeding and for right protection of a variety originator. In this reason attention is focused on the molecular markers generated by polymerase chain reaction. On the basis of STMS-analysis principles of identification and development of database, which reflect molecular-genetics peculiarities of some varieties of the Plant Breeding and Genetics Institute and other Ukrainian breeding organizations, are formulated. Allelic state at microsatellite loci and their distribution were investigated. Wheat varieties were ranged according to genetic distances, data on pedigree and cluster distribution of varieties obtained using computer programs were compared. PMID- 11944323 TI - [Genetic structure, variability and differentiation of Pinus sylvestris L. populations in the Ukrainian Carpathian Mountains and Rastoch'e]. AB - On the basis of electrophoretic analysis of 9 enzymous systems encoded by 20 gene loci the level of intra- and inter-population variation of two relict populations of Pinus sylvestris L. in the Ukrainian Carpathians and two ones in Rastochiye was studied. The less allele representation and the lower level of heterozygosity are typical for the Carpathian populations. Fst and Gst, parameters of populations subdivision, were not high--0.020 and 0.022 correspondingly and the coefficient DN was 0.008 in average. The results of the cluster analysis showed that only the populations of Rastochiye were united in one group indicating their genetic affinity. PMID- 11944324 TI - [Production of potato cybrids without using genetic selection markers of the parental material: Solanum tuberosum L. plants (cv. Svitanok Kyivsky) with S. pinnatisectum Dun. plastids]. AB - Interspecific cybrid plants of potato were created using the cell technology that does not require the presence of any genetic selectable markers in the parental material. Cybrid production was based on double inactivation of S. pinnatisectum Dun. protoplasts, served as the donors of organelles (by gamma-irradiation for the nuclei inactivation and by chemical mutagenesis for the efficient induction of chlDNA mutations), and the transfer of mutant plastids into S. tuberosum L. by protoplast fusion. Selection of cell colonies for streptomycin resistance was performed to identify the cybrid clones with mutant donor-type plastids. Restriction analysis of organelle DNA, chromosome counting, and isoenzyme analysis of the cybrids revealed the presence of nuclear material of S. tuberosum L. (cv. Svitanok kyivsky) and plastids from wild tuber-bearing S. pinnatisectum Dun. These plants enable the study of traits encoded by organelle DNA and to broaden the cytoplasmic diversity of cultivated potato. PMID- 11944325 TI - [Analysis of brain morphological changes induced by ethyl methanesulfonate in Drosophila melanogaster]. AB - Under the influence of ethyl methanesulfonate the series of both morphological and structural mutants with different types of brain changes has been obtained in Drosophila melanogaster Oregon R strain. In the future, this collection of mutants will be used in the investigations of genetic control of brain degeneration and possible ways of brain regeneration. PMID- 11944326 TI - [The spectrum of human chromosomal aberrations detected by routine and differential (GTG) staining]. AB - As a result of sample cytogenetic studies of 23 persons living on the territory of Yamal-Nentsy Autonomous District and chronically exposed to the small doses of radiation the data on the frequency and spectrum of chromosome aberrations, detected by the routine and differential (GTG) staining were obtained. Comparative efficiency of these methods was determined. The absence of significant differences for the spectrum and frequencies of chromosome aberrations revealed by both methods was shown. PMID- 11944327 TI - [Sex ratio in Down syndrome. Studies in patients with confirmed trisomy 21]. AB - Male to female ratio (sex ratio, SR) for 1,329 liveborns with Down syndrome and for 178,160 newborns from the general population of St. Petersburg, Russia was determined as a function of a mother age. Male prevalence (an overall SR of 1.24) was found in children with all trisomy 21 variants except the cases with mosaicism (the ratio of 0.88). The most expressed male predominance was determined in children of mothers aged 20-24 years, where SR was 1.73 in the total group (p = 0.00003) and 1.61 in the cases with free trisomy (p = 0.0007). Some hypotheses concerning the male accumulation in this group are discussed including a suggestion that the SR deviations from the population value 1.06 might be due to different contribution of paternal chromosomal non-disjunction during spermatogenesis. PMID- 11944328 TI - Chromosomal studies in infertile men. AB - Prometaphase and metaphase chromosome analyses performed on 70 consecutive men with primary infertility (for a period of at least 2 years) revealed 8 (11.42%) men with some kind of chromosomal abnormality. The highest frequency of abnormal karyotypes (10%) was found among patients with azoospermia and the most frequent anomaly was 47, XXY chromosomal constitution, found in 6 (8.57%) patients. All the chromosomal aberrations found in this study, was sex chromosomal type and we did not find any autosomal aberration. All patients with numerical chromosomal anomalies had azoospermia. The incidence of structural aberration in our study was 1.42%. 15 patients had different chromosomal variants (21.38%). We suggest that men with azoospermia should be considered for cytogenetic investigation and we report that "variants of the Y chromosome" have no influence on the sperm count (Million/ml) and fertility of men. PMID- 11944329 TI - [Structural and functional aspects of the nucleolar organizing region of interphase cell nuclei during hemoblastosis]. AB - Data on concept of the nucleolar-organizing regions of the human somatic cell interphase nuclei, methods of their revealing, classifications, as well as their structural-functional peculiarities in norm and under different diseases of blood system, in particular under hemoblastosis, are reviewed in historical aspect. PMID- 11944331 TI - [Oxidative activity and accumulation of metabolites in gonococcal and staphylococcal plasmid-containing strains]. AB - A higher rate of glucose oxidation by antibiotic-resistant staphylococcus and gonococcus strains has been established; at the same time the sensitive strains are characterised by more intensive oxidation of pyruvate and acetate as compared with the resistant strains. High rate of lactate accumulation by antibiotic resistant strains has been marked for the studied staphylococci strains. Antibiotic-resistant strains of gonococci are characterised by more intensive accumulation of acetate as the end product of glucose catabolism. No significant differences were revealed for the stable and sensitive strains of gonococci as to the rate of acetate involving in metabolic processes, but a steady tendency to the increase of acetate consumption intensity is observed. PMID- 11944330 TI - [Role of polypeptide growth factors in the regulation of epidermal keratinocyte proliferation]. AB - The growth factors, cytokines, adhesive molecules and extracellular matrix components play the leading role in the processes of intercellular interactions. Literary data on structure and mechanisms of functioning of the growth factors and their receptors are summarised in the present review. Some aspects of regulatory functions of such growth factors as EGF, TGFalpha, TGFbeta, FGF, KGF, AR, and HGF in these processes in epidermis keratinocytes both in vivo and in vitro as example were also considered. PMID- 11944332 TI - [Effect of bacterial physiological state and heavy metals on the hyperactive reaction of tobacco leaf]. AB - It has been shown that the initiation and proceeding of hypersensitive reaction on the tobacco leaf under the bacterial effect are influenced by the infectious loading of the pathogen, physiological state of bacteria, their viability, temperature of the environment and heavy metal salts. It has been established that the hypersensitive reaction is most distinctly manifested when 1-3 days bacterium cultures are used. Their activity decreases essentially with age, and 10-14 days old cultures are not able to evoke the hypersensitive reaction. The bacterial cells being inactivated with temperature and UV-irradiation lose the ability to evoke the hypersensitive reaction. The reaction of hypersensitive is inhibited by salts of heavy metals in concentrations 0.1-1% and by high temperature of the environment (30 degrees C). PMID- 11944333 TI - [Effect of various storage methods on the viability of myxobacteria]. AB - Viability of myxobacteria strains Myxococcus xanthus UNCM 10041, Polyangium cellulosum UNCM 10043, Archangium gephyra UNCM 10001 stored by the methods of lyophilization, cryoconservation, drying on paper discs, suspending in distilled water or physiological solution under a layer of mineral oil. The best results on viability preservation of all three studied strains were obtained when using the method of cryoconservation. It has been shown that the strains M. xanthus and A. gephyra preserve the best viability during two years of observations when stored in liquid nitrogen and dried on paper discs. Strain P. cellulosum preserves viability for two years when stored in liquid nitrogen and in suspensions on distilled water and physiological solution. PMID- 11944334 TI - [Discovery of the relationship between carbohydrate determinants of mollicute glycocalyx and affinity of microorganisms, and conditions for their existence]. AB - As a result of comparative analysis of interaction intensity of plant lectins, specific to certain monosaccharides with surface glycopolymers of philogenetically close groups of microorganisms--mollicutes and representatives of group Bacillus--Lactobacillus--Streptococcus the authors have established common features: availability of alpha- and beta-D-galactose, sialic acids, L fucose, N-acetylglucosamine, alpha-N-acetyl-D-galactosamine and alpha-D-mannose in their glycocalyx, that is a phenotypical manifestation of genetic relations between these microorganisms. The dependence between the availability of sialic acids and L-fucose in the composition of terminal structures of microorganisms glycocalyx and their capacity to occupy certain ecological niches have been found out. PMID- 11944335 TI - [Monoclonal antibodies used to differentiate between poliovirus types 1 and 3]. AB - A panel of mice hybrids was obtained, producing monoclonal antibodies (MCAB), which are capable to differentiate the vaccine and wild strains of polioviruses of the I and III types in the reaction of virus neutralization (RVN). The obtained MCAB activity was estimated on laboratory reference-strains and on the field isolates of polioviruses isolated in the territory of Ukraine in 1995-1998. The results of RVN with obtained MCAB were compared with the data of intratypical differentiation carried out with the use of standard MCAB from the Pasteur Institute as well as with results of the chain polymerization reaction (CPR) and immunoenzyme analysis (IEA) with cross-adsorbed sera. High correlation has been demonstrated between the results of the intratypical differentiation with obtained MCAB and data of the rest of the experiments. PMID- 11944336 TI - [Identification of plant viruses infecting transgenic potatoes resistant to the Colorado beetle]. AB - Virus tolerance of the three varieties of Bt-potato New Leaf--Atlantic, Russet Burbank and Superior from "Monsanto company" has been studied. Using serology, biochemistry methods and biotest the mixed virus infecting of experimental plants has been shown. Virus transmission by infected potato root crops under reproduction and accelerated degradation processes during storage have been registered. PMID- 11944337 TI - [Construction of the immunoenzyme test system "IFA-AtEBV-Strip" for detecting antibodies of the Epstein-Barr virus]. AB - The first home immunoenzyme test-system for finding antibodies to the Epstein Barr virus (EBV) has been created. The test-system laboratory prototype was called "IFA-AtEBV-strip". Production technology has been developed for specific components of immunoenzyme test system and all the analysis stage conditions have been optimized: the antigen sorption on plates, its interaction with the studied serum, finding out of the formed immune complex. The test system "IFA-AtEBV strip" sensitivity made up 96%, specificity--100%. PMID- 11944338 TI - [Interaction of microorganisms with solids]. AB - The paper generalizes data from literature dedicated to microorganisms interaction with solids. This interaction is widely spread in nature and considerably affects physiological activity of microbial populations. The role of physical, chemical and biological factors in this process is analyzed. It is noted that under certain conditions the hydrophobic forces are the basic factor in this interaction. The role of the cell surface components in this process is discussed. PMID- 11944339 TI - [Study of functional groups in the active center of alpha-galactosidase of Penicillium sp. 23]. AB - Effect of metal ions and specific chemical reagents (EDTA, sodium aside, o phenanthroline, n-chlormercurybenzoate, iodacetamide, N-ethylmaleinimide, L cystein, dithiotreitol, beta-mercaptoethanol) on activity of alpha-galactosidase isolated from the culture liquid of micromycete Penicillium sp. 23 has been studied. It has been established that alpha-galactosidase is not metalloenzyme (lack of the inhibitor effects under treatment of enzyme by EDTA, sodium aside, o phenanthroline). The heavy metal ions (Ag+ and Hg2+) inhibit the rate of alpha galactosidase reaction; Ki for Ag+ and Hg2+ ions makes up 3.3 x 10(-5) and 3.0 x 10(-7) M, respectively. It has been established by the inhibitory and kinetic analysis that a group (groups) with ionization constant about 6.0, are in the enzyme active centre which apparently corresponds to histidine imidasole group. The sulfhydryl groups do not take part in catalysis but play the important role in maintaining active conformation of the protein molecule. PMID- 11944340 TI - [Study of population dissociation of collection strains of Erwinia carotovora]. AB - Unusual dissociants of Erwinia carotovora subsp. atroseptica (EAT) population that emerge with high frequency (18-94%) during long-term storage of cultures were found. The dissociants (Poe-phenotype) differ from parental forms by slow growth and long development of the colour of colonies on EMB agar though they are not Lac-mutants. More than 60% of Poe-mutants of strain EAT 39A are characterized by 8- and 150-fold decrease of stability to erythromycin and oleandomicin, correspondingly. They are also sensitive to colicin-like carotovoricin, which is induced by mytomicin C, to carotovoricin of initial strain EAT 39A and to autocin, which they produce themselves during induction. A multiple phenotype of these dissociants (dissociants of the second type) is labeled: PoeOmSErSBnSAu. Its appearance as well as the appearance of pure Poe-variants is not associated with usual mutations and a loss of residential plasmids by cells. Population dissociants of the both types--Poe and PoeApSOmS--were also obtained from E. carotovora subsp. carotovora (J2) during growth at supraoptimal temperature. Poe variants are more often observed in representatives of subspecies atroseptica than carotovora and can be useful when studying pathogeneity of these practically important erwinias. PMID- 11944341 TI - [On the identity of Candida utilis and Pichia jadinii yeast species]. AB - Life cycles of Candida utilis (Henneberg) Lodder et Kreger-van Rij imperfect yeast (13 strains) used in industry were the study subject. When the strains were mated, we detected one of the stages of sexual process--conjugation of cells of the opposite mating type. Most of the studied cultures conjugated on the 2d-3d day. No ascospores were formed. Haploidy and heterothallism of the studied C. utilis strains were confirmed by hybridization of auxotrophic mutants. Based on PCR assay results, the yeasts are demonstrated to belong to the ascospore perfect yeast species of Pichia jadinii (A. et R. Sartory. We ill et Meyer, Wickerham) Kurtzman. PMID- 11944342 TI - [Investigation of subalin effect on urogenital microflora of pregnant women]. AB - Bacteriological examination of 30 women with incomplete pregnancy has been conducted. Representatives of 18 genera of aerobic and anaerobic bacteria as well as yeast-like fungi of Candida genus were found. It has been established that spore-forming bacteria of the Bacillus genus from subalin preparation have antagonistic activity in respect of the strains of pathogenic and opportunistic bacteria with characteristic zones of growth depression of 12-15 mm. This demonstrates that the agents of infectional urogenital process are effectively suppressed by subalin. PMID- 11944343 TI - [Specific immunoglobulins as effective drugs for treatment of virus infections: present state and urgent tasks in Ukraine]. AB - The thesis about importance of specific immunoglobulin preparations in treatment of infectious diseases of viral and microbial etiology, uncontrollable infections, caused by representatives of herpes viruses family in particular, is developed in the paper. The types of immunoglobulin preparations, their advantages, technology peculiarities has been considered; the experience of application, including that of the authors have been analyzed. The necessity and expedience of intensive development of this branch of immunobiotechnology in Ukraine is proved, especially in connection with a high level of infectious morbidity in this country, concrete recommendations are given. PMID- 11944344 TI - [Screening of collagenase and keratinase producers]. AB - The study of the capacity of 310 strains of microorganisms from different taxonomic groups (40 bacilli, 43 yeast, 105 streptomycetes, 12 micromycetes) to hydrolyze collagen and keratin allowed to establish that the highest level of collagenase (KA) and keratinase (KerA) activity is inherent in representatives of streptomycetes. Two strains of Streptomyces sp.--1349 and 1382 with the highest KA and KerA indices--1.9 and 1.85 un./mg of protein, respectively, have been chosen. It has been established that collagenase activity in the medium without adding the inducers decreases 4.76 times, while that of keratinase--5.71 times, i.e. the above enzymes are inducible. The investigation of the spectrum of activities has demonstrated that the both strains possess low level of the general proteolytic and elastase activities and high level of collagenase and keratinase activities. Partial purification of the enzyme complex of Streptomyces sp. 1349 by the successive precipitation by ammonium sulphate with 30, 60 and 80% saturation and a single precipitation by ammonium sulphate with 80% saturation helped to increase the level of KA 5.6-5.9 times, and that of KerA--4.2-4.5 times. PMID- 11944345 TI - [Chemical characterisation and serological activity of lipopolysaccharides of Ralstonia solanacearum]. AB - Lipopolysaccharides (LPS) of 8 strains of Ralstonia solanacearum (767, 4157, 5712, 7944, 7945, 7955, 8089 and 8110) characterized by different types of the structures of O-specific polysaccharide chains have been investigated. The ELISA and ELISA-inhibition methods were used to investigate the extent of antigenic activity of the presented LPS with respect to the obtained polyclonal O-antisera to the above strains. When analyzing the results obtained the strains were divided into five serogroups as the result of biochemical and serological investigations. The first serogroup included the strains 5712, 7945, 7955 and 8110, whose LPS and O-antisera were characterized by the high level of serological affinity. The second and the third serogroups included the strains 767 and 7944, respectively. Their LPS demonstrated antigenic specificity in the homologous systems only, as well as to O-serum to the strain 5712. The fourth and the fifth serogroups included strains 4157 and 8089, respectively, whose LPS did not manifest the antigenic specificity with respect to O-antisera of the other strains, besides their own ones, and which were characterized by different in principle types of the O-PS structures O-PS. PMID- 11944346 TI - [Ethanol formation by methane-utilizing bacteria at ethane co-metabolism]. AB - It was established, that EDTA (1.0 mM) and formamide (100 mM) are inhibitors of methanol dehydrogenase in Methylobacter luteus 12b, Methylomonas rubra 15sh and Methylococcus thermophilus 111p. The investigated strains co-metabolised ethane with the use of formate as the co-substrate. The application of formamide (or EDTA) as inhibitors of methanol dehydrogenase prevented from further transformation of ethanol and resulted in accumulation of extracellular ethanol. It was shown, that M. rubra 15sh accumulated extracellular ethanol under cultivation in a chemostate. The carried out researches have shown a regulation path of co-metabolism process of hydrocarbons by methane utilizing bacteria. Using the specific inhibitors of methanol dehydrogenase and a source of reducing agent (energy) for methane monooxygenase with the help of the cells of methane oxidizing bacteria it is possible to obtain from ethane or other hydrocarbons the products of their monooxygenation--alcohols. PMID- 11944347 TI - [Bsu50441--isoschizomer of endonuclease restriction AsuI]. AB - Endonuclease of restriction of type II has been found in Bacillus subtilis B-5044 during testing of endophytic strains of cotton plants bacilli. The restrictase Bsu5044I hydrolysed DNA of M13mp18 phage in 4 sites; pUC19 DNA in 5 sites; pBR322 in 15 sites. There were a lot of sites restriction in DNAs of phages T7 and lambda. A comparison of electrophoretical divisions of Bsu5044I and Cfr13I fragments of phages and plasmid DNAs showed their identity. Thus, the found restrictase Bsu5044I is an isoschisomer of AsuI. PMID- 11944348 TI - [Development of the method of RNA labeling in vitro using the terminal transferase with the purpose to obtain hybridization probes]. AB - A possibility to use terminal transferase to obtain hybridized RNA-probes has been investigated. Optimal conditions of the reaction proceeding were elaborated on the model objects--RNA of the phage MS2: 200 mM of potassium cacodylate, pH 7.2, 1 mM CoCl2, 1 mM DTT, under the considerable surplus of the enzyme. The elaborated method was used to determine the area of early genes of cyanophage LPP 3. With this purpose total RNA preparations were isolated from cyanobacterium Plectonema boryanum CALU 465. The above preparations were labeled by means of terminal transferase and [alpha-32P] ATP and used as the probes in hybridization with the DNA restriction fragments of LPP-3. Proceeding from the results of Southern-blots, a conclusion was made, that the KpnI-fragment of genome of cyanophage LPP3 10 kb in size includes the early genes. A possibility of practical use of the developed method is discussed. PMID- 11944350 TI - [Bacteriostatic action of fluorenylidene group derivatives on mycobacteria]. AB - Eight newly synthesized compounds--fluorenyliden derivatives have been studied for their antibacterial activity as to M. tuberculosis H37Rv and different kinds of atypical bacteria. The expressed antimicrobial activity in vitro of 5 [(fluorenyliden-3-hydrason)-aminomethyl]-thioglycolic acid (IR-95.6), lithium salt (fluorenyliden-5-aminomethyl)-thioglycolic acid (LI-15), lithium salt (fluorenyliden-9-hydraside)-thioglycolic acid (LI-75), with respect to M. tuberculosis H37Rv and considerable bacteriostatic action of all the studied substances as to M. fortuitum 23 have been established. PMID- 11944349 TI - [Serological affinity of some species of nonpathogenic corynebacteria]. AB - Serological peculiarities of the species strains Corynebacterium glutamicum, C. ammoniagenes, C. vitaeruminis, C. variabilis and strain of Corynebacterium sp. (Brevibacterium stationis) UCM Ac-719 have been investigated with the help of immunoenzyme analysis ELISA with the use of mice immune serum, specific to C. ammoniagenes UCM Ac-732T, C. vitaeruminis UCM Ac-718T, C. variabilis UCM Ac-717T, C. glutamicum UCM Ac-733 and Corynebacterium sp. UCM Ac-719. It has been established that the species of nonpathogenic corynebacteria differ between themselves as to the degree of serological affinity. C. variabilis, C. ammoniagenes and C. glutamicum are the least similar as to this indication. Weak antigenic relations have been revealed in C. vitaeruminis and C. ammoniagenes. The latter displayed the higher, as compared with other strains, affinity for Corynebacterium sp. UCM Ac-719. The highest degree of serological affinity within the species was registered in strains C. glutamicum and C. variabilis. Data obtained evidence that the ELISA method permits conducting the high-reliability species diagnosis of nonpathogenic corynebacteria on the basis of their antigenic characteristics. PMID- 11944351 TI - [Frequency of isolation of viable tuberculosis bacteria from patients with different forms of lung tuberculosis]. AB - Tuberculosis mycobacteria isolated from patients with lung tuberculosis have been analyzed to define their viability. The character of bacterial excretions was studied in patients with different clinical forms and detection genesis of tuberculosis process. During 1998-2000 about 228 cases of growth absence after mycobacteria inoculation have been revealed under positive microscopy. When determining viability of tuberculosis mycobacteria, it was shown that in most cases patients with all the forms of the pathologic process, except for the fibrous-cavernysh form, excrete alive microorganisms. The considerably higher amount of alive tuberculosis mycobacteria was distinguished in people who had chronical or relapsed tuberculosis in comparison with newly diagnosed patients. PMID- 11944352 TI - [Effects of nebivolol on serum levels of NO2 and NO3 oxides in patients with type 2 diabetes mellitus]. AB - In patients with type 2 diabetes mellitus, we used nevibolol, a lipophilic high selective beta 1-blocker. It has been found out that the drug, while augmenting the synthesis of nitrogen oxide, improves antioxidant properties. Therefore, it can be recommended for use in patients with type 2 diabetes mellitus concurrent with arterial hypertension to achieve normalization of arterial pressure and accomplish a correction of endothelial dysfunction. PMID- 11944353 TI - [Dynamics of morphological changes in tissues from the pancreas, liver, and the small intestine mucosa in acute pancreatitis]. AB - In acute cryogenic pancreatitis, apart from morphological changes in the affected organ there come to be seen destructive changes in tissues of the liver and small intestine that is evidenced by development in the liver of reactive hepatitis which is, strickly speaking, a result of a toxic effect of the pancreatic tissue necrotic products. In the small intestinal mucosa the above events are manifested by reactive enteritis, in which case both inflammatory-and-destructive and adaptive-and-regenerative processes are clearly seen. PMID- 11944354 TI - [Replacement of bone defects with a polyurethane composition with hydroxyapatite high-filling ]. AB - There have been obtained high-filling polyurethane compositions with hydroxyapatite. Compositions with hydroxyapatite and immunomodulator in the polyurethane base acquire better adhesiveness and improved strength characteristics as evidenced by physical-and-clinical studies made. Modal operations are developed in test animals on bone defects replacing that carry a high mechanical load. Histological studies showed that polyuretane-base high filling polymeric compositions replace the loss of bone fully, stimulate the processes of bone formation, with no negative responses of the osseous tissue noted. PMID- 11944355 TI - [Leukocytic component in disseminated intravascular blood coagulation syndrome]. AB - The experiments were carried out in nondescript rabbits. The animals were divided into two groups. Administered to both groups rabbits was the drug preparation "Efa-2"; in group II animals, inhibition of granulocytopoiesis was induced with myelosan. Group I rabbits developed hemostatic abnormalities characteristic of the subacute form of DIC-syndrome. Group II rabbits demonstrated hypercoagulation changes. In this way, inhibition of granulocytopoiesis and restriction of entering the blood of neutrophilic leucocytes lysosomal enzymes permit preventing the development of hemostatic disturbances characteristic of the DIC-syndrome by restricting the activity of both the sedimentation system and fibrinolysis. PMID- 11944356 TI - [Efficacy of combined herbal drug therapy in hypertension]. AB - The article contains a summary of the published literature together with results of the author's investigations designed to study efficacies of combined herbal drugs as treatment of hypertension. Treated at length are pharmacological mechanisms of action and a clinical effectiveness of a new combined vegetal drug chomviotensin that has come to be widely used as treatment of hypertensive disease. PMID- 11944357 TI - [Use of homeopathic remedies in the treatment of ulcer]. AB - The paper submits results of investigations designed to study efficacy of homeopathic medicines in treating peptic ulcer. Prospects are shown of employment of homeopathic therapy in modern treatment regimens for peptic ulcer. The place is substantiated by the authors of homeopathy in surgical, therapeutic, in patient, out-patient, sanatorium-and-health resort treatment settings. PMID- 11944358 TI - [Treatment of local hyperkinesis]. AB - Insufficient effectiveness of medicamentous therapy of local hyperkineses necessitated quest for other treatment options. In view of pathogenetic features of this pathological process we have chosen reflexotherapy to be instituted in our patients: auriculopuncture, electro-corporal- and acupuncture which have been shown to be expedient and effective in treating 65 patients with local hyperkinesis (blepharospasm, facial paraspasm, writer's spasm, spasmodic torticollis). Positive findings from clinical and paraclinical methods of investigation corroborate the results of the treatments administered. PMID- 11944359 TI - [Correction of the gastric and duodenal motility by laser puncture in pancreatobiliary diseases]. AB - 96.7 percent of patients with affections of organs of the pancreatobiliary zone displayed motor function disorders of upper portions of the alimentary canal (AC). A characteristic sign of the pathological process in pancreatobiliary organs is decreased frequency of recordable biopotentials and qualitative changes in electrogastrogrames. Changes in qualitative characteristics of the electrogastrogram are clearly related to increase in the intraduodenal pressure recordable with the aid of the "open catheter" technique. Laseropuncture is an effective supplementary method for correction of motility disorders in the upper portions of AC in those patients presenting with affections of the pancreatobiliary organs. PMID- 11944360 TI - [Use of local magnitothermia in the treatment of patients with primary-confined skin melanoma]. AB - We studied modifying potentialities of high temperatures in treatment regimens with adjuvant endolymphatic chemo- and interferontherapy of malignant melanoma of the skin. We had previously obtained enough evidence for expediency of adjuvant endolymphatic therapy of primary-localized melanoblastoma of the skin. Added to the preventive treatment regimens were sessions of local magnetothermia. The study was done in patients diagnosed as having primary-localized melanoma of the skin T(2-4) No Mo. A 3-year recurrence-free survival in the control group was (38.2 +/- 1.6)%, in the first study group--(54.2 +/- 5.8)%, in the second study group--(44.3 +/- 13.5)%. The general 3-year survival in the control group was (67.7 +/- 1.5)%, in the first study group--(70.8 +/- 5.5)%, in the second one- (91.7 +/- 5.7)%. In this way, the expediency has been proved of employment of local magnetothermia in treatment regimens to deal with malignant melanoma of the skin. PMID- 11944361 TI - [Mechanisms of therapeutic efficacy of magnetic laser irradiation of patients with neurological symptoms of lumbar osteochondrosis]. AB - Effects are studied of conventional treatments options and those used with a combined magnetolaser irradiation on clinical symptomatology, regional hemodynamics, and peripheral microcirculation in 254 patients presenting with neurological symptoms of lumber osteohondrosis. It is found out that employment of generally accepted treatment options permit achieving complete clinical remission in 38 percent of cases, with 28.7 percent improved, but parameters associated with hemocirculation fail to return back to normal. Medicamentous therapy and combined magnetolaser irradiation combined in treatment of the above bodily affliction have been shown to be conducive to clinical-and-neurological remission in 85.6 percent of cases, with the oxygen balance restored in all patients. PMID- 11944362 TI - [State of prostaglandin system in adolescents with the hypotensive type vegetovascular dysfunction during hyperbaric oxygenation]. AB - As many as 185 adolescents with hypotensive type vegetovascular dysfunction who ranged between 11 to 14 years old were studied for impact of a conventional treatment and that involving the use of hyperbaric oxygenation on the blood content of prostaglandins E2 and F2 alpha. The control group was 69 essentially healthy adolescents of the same age. Before the start of the treatment those persons presenting with hypotensive type vegetovascular dysfunction revealed an increase in prostaglandins E2 and decline in prostaglandins F2 alpha. There was no change in this trend with the conventional therapy. Owing to hyperbaric oxygenation, the above classes prostaglandins recovered their normal balance, which fact promoted reversal of clinical symptoms of the condition. The results secured permit recommending hyperbaric oxygenation as an efficient mode of treatment of hypotensive type vegetovascular dysfunction in juvenile subjects. PMID- 11944363 TI - [Characteristics of combined effect of thermal and welding aerosol exposure]. AB - While studying a concurrent exposure to thermal loads and welding aerosols (WA) it has been ascertained that in inhalational exposure of the body to WA under conditions of simultaneous exposure of animals to a high temperature of the air environment the total pathogenic effect tends to be higher and higher. It is suggested that in a concomitant exposure to the above factors WA might lower bodily resistance to the thermal load. The above factors produce summation type harmful effects which have clearly a dose-dependent character. Studies on the occupational environment of workers engaged in welding involving heating suggested to us that it is those WA containing harmful substances of I and II hazard class that is the leading adverse factor of the working environment, and also thermal loads as high as 8000-9000 wt/m2. A concurrent exposure to the above factors lays a stress on the thermoregulatory function of the organism, and is resultant in the development of autoimmune processes and in premature aging of workers. PMID- 11944364 TI - [Compartment syndrome in pathogenesis of lower limb exertional disease in servicemen]. AB - As many as 35 servicemen presenting with a pathological reorganization and stress fractures of shin bones and foot were examined. A measurement of the subfascial pressure was done together with a study of pulse propagation along the tibial and fibial nerves and of bioelectrical activity of foot muscles innervated by the above nerves. The subfascial pressure in the anterior and posterior deep sheaths of the shin appeared to be increased in rest, especially so in the wake of physical loads on the affected extremity even if the process was localized in the foot. The velocity of the pulse propagation along the nerves was found to be decreased, the latent period increased, amplitude decreased, the duration of M response longer in the affected extremity, with a 38 to 47% decrease in the frequency and amplitude of biological activity of maximum muscular contraction having been recordable. The latter observation has corroborated our assumption that in weight-bearing disease of the extremities there would be present a chronic compartment-syndrome which is of reccurrent character, causes disturbances in the neuromuscular apparatus of the extremity (ischemic demyelinization of the nerve fibers and dystrophic changes in the muscles innervated by the above nerve fibers), which fact may lead to recurrences of weight-bearing disease, prolonged treatment and restoration of the function. PMID- 11944365 TI - [Analysis of suicides committed by employees of the internal affairs bureau while in a state of alcoholic intoxication]. AB - The article analyzes proceedings in the inquiry of 93 cases of suicide that IAB officials have committed in the state of alcohol intoxication. PMID- 11944366 TI - [Pathogenic characteristics of osteoporosis in patients with rheumatoid arthritis]. AB - From the modern standpoint, interrelation is shown between different mechanisms of osteoporosis (OP) and course of rheumatoid arthritis (RA). Allowing for a large variety of mechanisms of development of OP in RA, especial emphasis is placed on identifying pathogenetic variants of development of OP in RA. The available data from the published literature indicate further study for the role of cytokins in the development of OP in RA. PMID- 11944367 TI - [Public relations in institutions and establishments of the health administration system]. AB - The article is dedicated to development of directions and specific functions of the health system bodies/institutions public relations (PR) activities. Priorities are set forth depending on the form of property thereof. A complex use of approaches toward carrying out of PR activities permits optimizing work both within the system itself and relations with the society as a whole. PMID- 11944368 TI - [Improvement of the prehospital care for patients with myocardial infarction]. AB - The article highlights objectives and tasks of the emergency cardiological service at the prehospital stage. Recommendations are given on improvement of diagnosis and organization of medial service delivery to patients with myocardial infarction. An association has been demonstrated between the clinical picture of myocardial infarction and duration of hospitalization. The possibility is considered of diagnosis of myocardial infarction in atypical clinical picture at the stage of emergency medical care to be provided for the patients under consideration. PMID- 11944369 TI - [Prevention of sudden infant death syndrome in children in preventive-treatment institutions in the Odessa region]. AB - The sudden infant death syndrome (SIDS) is one of the most prevalent causes of infant mortality in European countries. SIDS is a sudden but not violent death in those infants aged between 7 to 365 days. Whether or not the case can be diagnosed as having had SIDS remains uncertain even after an in-depth study of the autopsy material, previous history, and death concurrent events. The study into infant care was made within the framework of the ICCPS (International child care practical study) program with 21 countries from all the corner of the globe having taken part in it. SIDS incidence in Ukraine estimated with the aid of the model of the Odessa Region has been found out to come up to 0.5-0.85%. Measures have been instituted designed to prevent SIDS as a stage of our work at women health centres, mother and child outpatient and in-patient departments of the relevant institutions. Monitoring of children at risk for SIDS is not a sufficient enough measure capable of reducing SIDS incidence as evidenced by a one-year follow-up but combined with the parents' education as to risk factors for SIDS in a preventive-treatment setting such a monitoring permitted reducing SIDS frequency from 0.85 to 0.60% in the Odessa Region. PMID- 11944370 TI - [The thermal pattern of skin in healthy persons]. AB - Based on data from published literature the author has reached the conclusion that the thermal pattern in healthy people has many "hot" and "cold" zones the occurrence of which is due to exposure to a number of anatomical and physiological factors, in the first place,--to particular features of blood supply. However, the distribution of infrared radiation relative to the middle line is in fact symmetrical, the physiological temperature difference does not exceed 0.5 degree C. The knowledge of normal thermal patterns is the necessary condition in interpreting the thermogram in the patient. PMID- 11944371 TI - [Immunomodulating effect of berlithione in clean-up workers of the Chernobyl nuclear plant accident]. AB - Effects of the antioxidant berlithione (alpha-lipoic acid) on indices for the immune system, such as complement titer, C3-component of the complement, reaction of the autorosette formation were studied together with those on parameters characterizing cell-bound and humoral immunity. 11-12 years following the Chernobyl accident, nine liquidators of its aftermaths were examined, aged 51.8 +/- 7.2 years, body mass (BM) 27.3 +/- 4.2 kg/m2, radiation dose 73.2 +/- 35.3 rem, who took part in the elimination of the effects of the accident for no more than 3 months in 1986--at the beginning of 1987. All examinees were prescribed the drug berlithione (alpha-lipoic acid), 600 mg per day over two months. A two month treatment with berlithione, 600 mg/day, resulted in normal levels of indices for unspecific defence; among these were phagocytic activity of neutrophiles, content of the complement, its C3-component, and reaction of autorosette formation in the accident effects liquidators, but no impact was found on cell-mediated and humoral immunity. PMID- 11944372 TI - [Effect of low intensity electromagnetic waves from cell phones on human health]. AB - Data on the negative effect of low-intensity electromagnetic waves on organs and systems of the human body are submitted together with results of studies into efficiency of use of protective devices. PMID- 11944373 TI - [Prospects in the treatments of stomach neoplasms]. AB - Highlighted in the paper are basic treatment modalities for carcinoma of the stomach at the present stage of development of abdominal oncology, including original developments of the Ukrainean Research Institute of Oncology and Radiobiology of the Ministry of Health of Ukraine as the head institution of the country. The main index of references reflect potentialities of and prospects for treatment of gastric carcinoma over the last ten years. PMID- 11944375 TI - [Pain sensitivity threshold in patients with ischemic heart diseases with and without stable angina with episodes of "silent" myocardial ischemia]. AB - A total of 124 coronary patients were examined. Group I comprised 52 patients with stabile angina, group II was 72 patients free from angina. In all patients, 24-hour ECG monitoring was carried out together with veloergometry (VEM), coronaryangiography. Determined in the above patients was also threshold of pain sensitivity (PST) by using the precardiac area electroskin stimulation technique. The frequency of pain-free episodes of myocardial ischemia (PFMI) in subjects with stabile angina has not been found to be different from that in those free from it. There was no difference in depth and duration of depression of ST segment either. No significant differences were found out between PFMI and pain episodes of myocardial ischemia in group I subjects. PST in both groups subjects was not associated with the number of damaged coronary arteries (CA) and presence of the asynergic zones in the left ventricle (LV). In angina-free subjects presenting with the single CA affection, the right coronary artery (RCA) appeared to be affected much more frequently than in those patients with stabile angina. No primary damage to RCA could be seen in associated CA afflictions. PMID- 11944374 TI - [Epidemiology of thyroid diseases in iodine deficiency]. AB - Prevalence was studied of thyroid pathology under conditions of a moderate iodine deficiency in the Kolomyya District of Ivano-Frankovsk Region. Both pediatric and adult population (n = 1245) was examined, residing in the villages of Markivka and Rungury. The prevalence of diffuse goiter among children less than 14 years of age came up to 63.3% falling to 24.2% among those persons aged 56 to 75 years. In the 36 to 55 age bracket, thyroid nodularity has been found to be on the increase. The incidence rate of thyroid carcinoma in the examined population has come to be 0.1%. Children and juveniles are especially vulnerable to iodine deficiency since it is these age populations that have much higher incidence of diffuse goiter. PMID- 11944376 TI - [Effect of beta-blockers on the cardiac rhythm variability in patients with hypertension]. AB - As many as 29 patients with arterial hypertension were examined. All patients were instituted a 3-week antihypertensive therapy with metoprolol. Against the background of intake of metoprolol, there occurs a rise in indices for variability of the heart's rhythm. Augmentation of total power of the spectrum due to metoprolol occurs largely because of enhancement of the parasympathetic activity which can be regarded as a positive effect on the sympathicopara sympathetical control. In apparent enhancement of the parasympathetic nervous system against the background of ingestion of metoprolol the activity of the sympathetic component is not inhibited, the drug leading to redistribution of influences toward that portion of the autonomic nervous system which had been more inhibited before the intake of the drug. PMID- 11944377 TI - [Dysplastic cardiopathy concurrent with the heart rhythm and conductivity disturbances in adolescents]. AB - Clinical-and-hemodynamic features have been studied of dysplastic cardiopathy presenting with disturbances of the heart's rhythm and conductivity in 90 boys and girls 12-18 years old. It has been ascertained that high degree of the mitral valve prolapse, multiple abberrant chords, local hypertrophy of the left ventricle are predictors of development of arrhythmias in juveniles with dysplastic cardiopathy. The intracardiac hemodynamics has been found to be affected by "small" cardiac stigmas of dysembryogenesis. There was noted a decline in the heart's pumping and contractile functions resulting in formation of hypokinetic type hemodynamics. The findings secured attest to the need for a case follow-up of those patients who are at high risk for development of arrhythmogenic cardiomyopathy presenting with development of irreversible or refractory to correction complications. PMID- 11944378 TI - [Determination of differentially amplified ECG indices for the ventricular complex in healthy children and in children with heart and rheumatic diseases]. AB - As many as fifty healthy children and juveniles who ranged between one to 18 years old were examined together with 25 children presenting with preexcitation syndromes (WPW-syndrome). Registered in all patients was the DAECG tracing. As a result of the studies made it has been found out that indices for the DAECG ventricular complex are age-dependent. DAECG indices characterizing WPW-syndrome are defined. PMID- 11944379 TI - [Changes in the vegetative status and excretion of catecholamines during lateral therapy for essential hypertension]. AB - In 30 patients with stage II essential hypertension a spectral analysis of the heart's rhythm was done together with measuring the urine excretion of catecholamines before and after a session of lateral light therapy. There has been noted a decrease in power of the ranges 0.08-0.15 and 0.15-0.5 Hz of the spectrogram and an appreciable rise in excretion of dofamine. The investigators have reached the conclusion that under the exposure to lateral light therapy there occurs a decrease in the baroreflex and vagus tone. A suggestion is made to the effect that dofamine may have its part in the antihypertensive effect of lateral therapy. PMID- 11944380 TI - [Comparative analysis of clinical computer electroencephalographic findings in patients with acute cerebral hemispheric and hemorrhagic stroke]. AB - As many as 146 patients in the acute period of cerebral ischemic insult (CCI) and cerebral hemorrhagic insult (CHI) of hemispheric localization were examined. Clinical-and-neurological examination was done together with that computed tomography (CT)-guided, computerized electroencephalography (CEEG), applied mathematical statistics. Polymorphous relation has been revealed between the clinical pattern of cerebral insult and CT/CEEG criteria. The strongest relation has come to be between the localization and clinical pattern of CCI, and between the size of the affected area and clinical patterns of CHI. Parameters of the EEG pattern closely correlate with clinical patterns of insult. PMID- 11944381 TI - [Treatment of epileptic seizures resistant to phenobarbital in children during the first year of life]. AB - In a prospective investigation, efficacy and safety of use of valproate (depakine syrup) was studied in infants presenting with the epileptic convulsive syndrome that was resistant to phenobarbital. The study comprised 26 patients who ranged from 3 months to 1 year old presenting with primary and secondary generalized convulsive epileptic fit. The use of valproate (depakine-syrup) in a medium therapeutic dose (15 +/- 5) mg/kg per day permitted controlling fits in sixteen babies (61.53%) over six weeks; in other three infants (11.53%) the frequency of paroxysms has come to be strikingly decreased, in one infant (3.84%) there has been achieved only partial control of fits, and it is only in 6 babies that the state remained unchanged. In this way, the use of valproate (depakine-syrup) in babies presenting with primary and secondary generalized cerebral seizure resistant to phenobarbital is an effective means of control of seizures with lesser incidence of ill effects recordable than in case of employment of phenobarbital. PMID- 11944382 TI - [Various mechanisms of cytoprotective effect of omeprazole and low intensity laser radiation on the gastroduodenal mucosa in the treatment of patients with duodenal ulcer]. AB - Clinical studies were made in 130 patients with duodenal ulcer in the phase of exacerbation of the disease. There were 98 men and 32 women who ranged from 17 to 50 years old. Results of examination of 7 essentially healthy subjects were regarded as control. The patients were divided into three groups. Group I patients (n = 48) received a conventional therapy; in group II patients, the adopted therapy was supplemented by omeprazol, 20 mg twice daily, group III patients (n = 43) were (in addition to the above therapeutic regimen) exposed to a session of endoscopic low-intensity laser irradiation (LILI) for 5 min (overall 6 to 8 LILI procedures). It has been ascertained that omeprazol exerts a cytoprotective effect on the mucozal barrier of the gastroduodenal zone brought about by increase in the synthesis of glucoproteins in the mucous membrane, improvement of the water-and-elastic properties, and enhancement of resistance of the mucosal barrier to the action of the aggressive factors. Administration of endoscopic LILI treatments in DU patients has also been found out to have a cytoprotective effect but superior to omeprazol. A protective action of LILI is believed to be caused by stimulation of synthesis of the most important components of glycoproteins. A cytoprotective effect of omeprazol and endoscopic LILI is ccompanied by a significant shortening of time for the clinical symptoms to get dispelled, the ulcer cicatrization frequency increased. PMID- 11944383 TI - [Clinical significance of serum levels of medium size molecules in patients with iron deficiency anemia]. AB - Overall 45 patients (20 men and 25 women) with iron deficiency anemia (IDA) were examined. The content was measured of medium-size molecules (MSM) in the blood serum of patients with different degree IDA. The time-related course was followed of changes in MSM content during the course of treatment of the patients. The control group comprised 22 essentially healthy subjects. The blood serum content of MSM in IDA patients proved to be significantly higher as compared to healthy individuals (P < 0.001). Recordable in the patients was a significant IDA severity-related increase in MSM content. Clinical, diagnostic, and prognostic significance of MSM content in IDA patients is discussed. It is suggested that the foregoing biochemical criterion be used in a comprehensive assessment of degree of severity of the endogenous metabolic intoxication in patients with IDA. PMID- 11944384 TI - [Perioperative hypotensive therapy of patients with chronic renal insufficiency]. AB - Overall 281 patients with arterial hypertension (AH) and renal insufficiency were examined, who had been operated on for renal diseases and those of the urinary tract. It is ascertained that patients with similar degree of AH can have both hyperkinetic and hypokinetic type of circulation. It is shown that most patients do not receive regular antihypertensive therapy before their admission into the hospital and are in want of its institution during the process of the preoperative preparation. Variants are proposed of prescription of hypotensive drugs with special reference to the type of circulation in the patient and to the chosen method of anesthesia. The conclusion in that a rational selection of antihypertensive therapy and optimum method of anesthesia ensure stability of hemodynamics in the perioperative period. PMID- 11944385 TI - [Prophylaxis for infectious complications of transurethral resections for prostatic hyperplasia]. AB - A clinical-and-laboratory analysis was done in 76 patients who had undergone transurethal resection (TUR) for benign hyperplasia of the prostate (PH). Included into a complex clinical-and-laboratory examination were bacteriological urine tests, with the causative agent isolated and identified, analysis of the course of the affection, assessment of X-ray findings, clinical analyses of urine and blood. With the purpose of preventing pyo-septic complications before surgery and postoperatively, cephtasidim (fortum), a broad-spectrum antibiotic, was prescribed. The analysis of the clinical studies made suggests to us that a rational antibiotic prophylaxis involving the use of cephtasidim in those patients presenting with benign PH makes the risk of development of the post-TUR PH infectious complications getting lower. PMID- 11944386 TI - [Changes in the local proteinase-inhibitor system in children with chronic erosive gastritis]. AB - In 26 children presenting with chronic erosive gastritis, an unspecific proteinase-inhibitor potential was identified in the basal portion of the gastric juice in the period of exacerbation and incomplete remission. Revealed in the gastric content in the period of exacerbation was augmentation of the activity of elastase- and tripsinlike proteinases. In the period of remission, the indices have been shown to be lower but insignificantly, remaining significantly higher compared to those indices in the control group. There was a decrease in the antiproteinase potential in the period of exacerbation getting even more lowered during the period of incomplete remission. The findings secured suggest to us a manifest imbalance in the local proteinase-inhibitor system. A conclusion has been reached that exhaustion of the local inhibitor potential and ensuing high activity of unspecific proteinases result in destructive changes in the gastric mucosa, thus, having pathogenetic significance in the development of chronic inflammatory-and-destructive diseases. PMID- 11944387 TI - [Neurologic status of children with chronic hepatitis B]. AB - In those patients presenting with chronic virus hepatitis B (ChVH B), it is not only the liver that is involved in the pathological process but also the vegetative nervous system, its condition being dependent on the activity of the process in the liver. PMID- 11944388 TI - [Effect of various methods of functional osteosynthesis on healing of the femoral diaphysis fractures in adults]. AB - Our objective in this investigation was to study healing of fractures of the femur diaphysis treated with different methods of fixation. A total of 177 patients who ranged from 16 to 80 years old were examined. It has been ascertained that union is through secondary callousity formation. Our techniques for screw adjustment involving the use of a dynamic compression plate and intraosseous nail made for a complete consolidation of fractured bone fragments within one year. The use of a plate with screws arranged at an interval of up to 1 sm near the plane of the fracture or external fixation unit prolonged healing time. Recommendations have been worked out in the course of the study to improve methods of treatment of fractures of the femur diaphysis. PMID- 11944389 TI - [Management policy for osteoarthritis of the knee joint]. AB - Osteoarthrosis of the knee joint (KJA) is a most prevalent orthopedic disease. There are some questions as to the policy of treating KJA that remain to be decided. Results are analyzed of treating 3243 patients with KJA (stage I--889 individuals, stage II--1276 ones, stage III--1078 patients) over the period 1983 2000. The patients underwent nonoperative treatment, arthroscopic operations, plasty of the ligamentous apparatus, osteotomy, arthrodesis, total endoprothetics. Our scheme proposed some time ago allowed judgement about the function of the knee joint. Recommendations are given in the article, that include the following: Arthroscopic operations should be widely used during stage I and II for the debridgement of the joint to be done, the proper recovery of the ligamentous apparatus to be ensured. Correcting osteotomy is indicated in axial deformity of the joint at stage I to II. Total andoprothetics is indicated at stage III osteoarthrosis and also at stage II in case of joint instability and axial deformity. PMID- 11944390 TI - [Apoptosis in myocardial ischemia and reperfusion]. AB - Apoptosis initiated in cells of tissues maintaining homeostasis under the influence of growth factors and hormones has been recently described during the early stage of myocardial infarction in the reperfusion zone of the cardiac muscle. Although it has been proved that apoptosis of cardiomyocytes plays one of the key roles in progression of the reperfusion syndrome, molecular and cellular mechanisms of this phenomenon are not yet resolved. Prospects are discussed of a new trend in preventive treatment of ischemic and reperfusion-related affection of the myocardium relying upon blocking of apoptosis of cardiomyocytes. PMID- 11944391 TI - [Treatment of dystrophy, dysplasia and initial stages of vulvar carcinoma in virus infections]. AB - Overall sixty-seven patients with distrophy, dysplasia and valvular carcinoma presenting with human papilloma virus- and herpes simplex-2 virus infections underwent a complex treatment intended for background, precancerous affections of the vulva and for valvular carcinoma, involving the use of antiviral drug preparations laferon, zovirax, lovir (both on the systemic and local basis), and an immunomodulator (decaris). Efficacy of the above treatment option has been shown to be 98.5%. PMID- 11944392 TI - [The crystal-optic method as an objective criterion for assessment of the course of the wound process in parturient women]. AB - The structure was studied of biological media (blood, wound exudation, biopsy specimen from the wound) in 119 parturient women presenting with suppurating wounds to diagnose the course of the wound process, using the method of polarization microscopy. The biological material was taken before the treatment, during its course, and after the treatments administered. Based on the studies made the authors have come to the conclusion that the crystallooptic method reflects the bodily state of the women in labour validating the pathogenesis of the pyoinflammatory diseases; it is helpful in following the change of phases of the wound process, showing efficiency of treating purulent wounds, rejuvenation of tissues in the wound toward their primary functional state. PMID- 11944393 TI - [Meckel phlegmonous diverticulum concurrent with gangrenous inflammation of the vermiform process]. AB - Historical data on the Meckel's diverticulum are presented together with the analysis of the latest published literature data. A case is described of concomitant Meckel's flegmonous diverticulum and gangrenous appendicitis in a child eight years old with the clinical picture outlined at the time of admission into the hospital and surgical and conservative treatments administered. PMID- 11944394 TI - [Protracted atypical course of enterovirus infection]. AB - Two cases are presented of replication of enteroviruses over a long period of time, that caused a protracted atypical clinical course of infection. No typical clinical markers of infection can be seen in the wake of enterovirus replication; the replication itself can be of a protracted recurrent character. Replication of enteroviruses is capable of inducing a protracted recurrent subfebrile condition in adults that has no clinical signs of immunodeficiency state. Parallel activation of replication of a number of viruses, according to our data, is a widespread phenomenon, which fact is to be taken account of in planning examination and treatment strategies. Laboratory tests are needed for the activation of diseases of enteroviral etiology to be ascertained. PMID- 11944395 TI - [Psychiatric aspect of Gilles de la Tourette disease]. AB - A case of Gilles de la Tourette's disease of adult onset is described. An analysis of time-related course of mental derangements is presented. PMID- 11944396 TI - [Gastrozepin therapy of gallbladder motility disorders in patients with ischemic heart disease]. AB - Submitted in the paper are results of studies on gastrocepin effects on degree of clinical symptoms, contractility of the gall bladder, and power of the threshold load in patients with ischemic heart disease and disordered motor function of the bile tracts. Gastrocepin is shown to increase contractility of the gall bladder and facilitate normalization of the Oddie's sphincter tone; it makes for the reduction in the incidence of anginal attacks and increase in the physical exercise tolerance in patients with stabile functional class I and II angina pectoris of effort. PMID- 11944397 TI - [Use of pentoxifylline in the treatment of systemic scleroderma]. AB - Analyzes in the paper are chief pharmacological effects of pentoxifylline; the necessity is substantiated of its prescription in a combined therapy of systemic scleroderma (SSD). As many as 220 SSD patients on an in-patient care were examined. Apart from general clinical, biochemical, immunological methods of investigation, the condition of the microcirculation system was studied, the level of the tumor necrosis serumal factor alfa (TNF-alpha) was measured. The data secured suggest a considerable increase in the level of TNF-alpha and pronounced disturbances in microcirculation. The administration of pentoxifylline to SSD patients has been found to improve the microcirculation and reduce the production of the above factor, which fact suggests its immunomodulating properties. PMID- 11944398 TI - [Different concepts....in comparison]. PMID- 11944399 TI - [Violence against women. Looking the facts in the eye]. PMID- 11944400 TI - [The Swiss Qualifications Manual CH-Q. Making hidden abilities visible]. PMID- 11944402 TI - [Practicum in the Philippines. Everything is completely different]. PMID- 11944403 TI - [Imprisoned in the norm]. PMID- 11944404 TI - [Measuring steps, taking time]. PMID- 11944401 TI - [European model for total quality management (TQM). A method for self evaluation]. PMID- 11944405 TI - [Victims]. PMID- 11944406 TI - [The Romeurope Project. The health of the Roms in a situation of total exclusion]. PMID- 11944407 TI - [A Swiss minority without territory. Behind the sign of travel]. PMID- 11944408 TI - [Emergency program in Romania. Free health care for inhabitants of Zabrauti]. PMID- 11944409 TI - [Health professionals faced with violence. "Your jaw bone is fractured. How did it happen?]. PMID- 11944410 TI - [Personnel development against professional power. Self development to better prevent burnout]. PMID- 11944411 TI - [The art ofof high wire dancing]. PMID- 11944412 TI - [Stress management. "Imagine a river..."]. PMID- 11944413 TI - [N as in need]. PMID- 11944414 TI - [Nursing relations. Perceiving and feeling the proper distance]. PMID- 11944417 TI - [The future of nursing care. "So our fate isn't like that of the Indios..."]. PMID- 11944418 TI - [Project of the Bern SBK section. "House of nursing"]. PMID- 11944419 TI - [Social phenomenon. Violence in the family--what should be done?]. PMID- 11944420 TI - [Intimacy--also in advanced age]. PMID- 11944421 TI - [Reintegration course. Personality development is the most important step]. PMID- 11944422 TI - [With dignity]. PMID- 11944423 TI - [Families with HIV-/AIDS problems need professional support]. PMID- 11944424 TI - [Active waiting]. PMID- 11944425 TI - [Training on the move]. PMID- 11944426 TI - [Public health universities of Switzerland and training in nursing care. The work progresses]. PMID- 11944427 TI - [Training at the university II. Nursing care aid specializing in health]. PMID- 11944428 TI - [Evaluation of students in nursing care. The status of school-training collaboration]. PMID- 11944429 TI - [A new model of clinical teaching. When change rhymes with adjustment]. PMID- 11944430 TI - [H as in hope]. PMID- 11944432 TI - [Multi-ethnic nursing. "Cross-cultural care: an emerging necessity]. PMID- 11944433 TI - [The way back into the future]. PMID- 11944434 TI - [Sleep difficulties. There is more available than reaching for the drug cabinet]. PMID- 11944435 TI - [Discharge planning. Termination of a process]. PMID- 11944436 TI - [Transplantation coordinator. A permanent challenge]. PMID- 11944437 TI - [Group meetings. The deciding factor is reflection]. PMID- 11944438 TI - [Half the Easter egg]. PMID- 11944439 TI - [Child-like spontaneity as the key]. PMID- 11944440 TI - [An extraordinarily ordinary journey]. PMID- 11944441 TI - [Understanding a diffuse handicap. Aphasia or the lost words]. PMID- 11944442 TI - [How to communication with an aphasic person? Strategies and attitudes to adopt]. PMID- 11944443 TI - [United Nations 2001: year of benevolence. The benevolent: a reputation to defend]. PMID- 11944444 TI - [Interview with Ruth Dreiffuss. "No community can do without benevolent work"]. PMID- 11944445 TI - [Exposition at CHUV. THe indispensible hut]. PMID- 11944446 TI - [Caring for a patient with a stoma. Living with a stoma: what is impossible becomes possible]. PMID- 11944447 TI - [Complementary methods]. PMID- 11944448 TI - [Valuing the conscientiousness of nurses. Freeing ourselves from our history!]. PMID- 11944449 TI - [G as in grief]. PMID- 11944451 TI - [Oral hygiene. Water and tea: effective without side effects]. PMID- 11944453 TI - [Aphasia. When words fail]. PMID- 11944454 TI - [Opening address. Family caregivers: what challenges for the future?]. PMID- 11944455 TI - [Compliance. Methods for better cooperation]. PMID- 11944456 TI - [A day unlike any other]. PMID- 11944457 TI - [Finger Food. Eating independently with pleasure again]. PMID- 11944458 TI - [Taming the present]. PMID- 11944460 TI - [Annual Congress of ASI in Zurich. Venture forth!]. PMID- 11944459 TI - [From technique to emotion. Death and rituals in intensive care]. PMID- 11944461 TI - [World Health Assembly. "The nurses have drunk the whey"!]. PMID- 11944462 TI - [Nursing opinion. "Real elan for changing our image"]. PMID- 11944463 TI - [Blasting the comfortable framework apart]. PMID- 11944464 TI - [A great women in nursing care retires. Homage to Kay Carpenter]. PMID- 11944466 TI - [Respecting the autonomy of the child]. PMID- 11944467 TI - [G as in gamin]. PMID- 11944469 TI - [SKB Congress 2001 in Zurich. Dropping the old, having new ideas]. PMID- 11944471 TI - [Early complications of surgery for chronic suppurative otitis media]. AB - OBJECTIVE: To explore early post-operative complications following surgery for chronic suppurative otitis media. METHOD: From Jan 1992 to July 2000, 291 cases of chronic suppurative otitis media have been treated in our department. 79 cases underwent myringoplasty, 82 cases tympanoplasty, 88 cases modified radical mastoidectomy and 42 cases radical mastoidectomy. RESULT: Facial nerve palsies occurred in 3 cases (1.03%), bone conduction threshold elevation occurred in 13 cases (4.47%), wound infection occurred in 11 cases (3.78%), 7 patients (2.41%) reported symptoms related with chorda tympani trauma, symptoms of jaw discomfort were reported by 7 patients (2.41%) and imbalance or vertigo by 13 patients(4.47%). CONCLUSION: The incidence of early complications of surgery for chronic suppurative otitis media in this report was similar to previous reports. PMID- 11944472 TI - [Analysis on clinical data of 677 death cases with nasopharyngeal carcinoma]. AB - OBJECTIVE: To study the death causes in 677 patients with nasopharyngeal carcinoma (NPC). METHOD: The clinical data of 677 death cases with NPC hospitalized in our department from 1974 to 1990 were analysed. RESULT: In our data, the ratio of male to female was 4.3:1. The main age stage was from forty to sixty. The pathological type was mostly poorly differentiated squamous cell carcinoma. The cases with stage III-IV disease were account for 81.3%. The patients treated by radiotherapy combined with chemotherapy survived longer than those treated by radiotherapy alone (P < 0.05). The majority of the patients died of distal metastasis (372, 54.9%) and local-regional lymph node uncontrolled (142, 20.9%) after treatment. CONCLUSION: The main death causes of the patients with NPC in our investigation were distal metastasis and local-regional recurrence. The treatment of radiotherapy plus chemotherapy probably increases survival time and reduces the death rate in patients with NPC. PMID- 11944473 TI - [Endoscopic anatomy of the middle ear]. AB - OBJECTIVE: To provide a detailed anatomic description of middle ear endoscopy via different approaches. METHOD: Using 0, 30 and 90 degree rigid endoscopes, 6 dry temporal bones and 11 temporal bone blocks were examined to define the average range of view obtained by three approaches: 1. via the external ear canal approach; 2. via a retroauricular incision: including two subdivided routes, the mastoid-tympanic antrum route and mastoid-retrotympanum route; 3. via the middle ear cranial fossa. Each region of the middle ear was chosen for inspection. RESULT: 1. via the external ear canal approach, all sites of the tympanic cavity structures were seen with 0, 30 and 90 degree rigid otoendoscopes, including the hidden areas such as the tympanic sinus, facial recess, anterior epitympanic recess, orifice of eustachian tube et al. 2. via the mastoid-tympanic antrum route, the structures of the mastoid cavity, tympanic antrum, epi-mesotympanum were clearly defined; via the mastoid-retrotympanum route, the structures of the mastoid cavity and meso-hypotympanum were also observed; 3. via the middle cranial fossa approach, the structures of the epitympanum, tympanic antrum, mastoid cavity and lateral wall were inspected, most mesotympanic structures were evaluated. CONCLUSION: Middle ear endoscopy through the different approaches provides the otosurgeons a clear view field of the middle ear structures, can therefore decrease the incidence of cholesteatoma recurrence and evaluate the state of the postoperative middle ear cavity. The 30 degree otoendoscope can provide a better visualization of the middle ear structures, via the three approaches, the 90 degree otoendoscope is suited for the inspection of the posterior epitympanum via the transmeatal approach and retrotympanum via mastoid cavity-tympanic antrum route. PMID- 11944474 TI - [Effect of hyperlipidemia on auditory brain stem responses]. AB - OBJECTIVE: To investigate the effects of increasing stimulus rate (ISR) of sound stimulation in ABR test and obtain experimental data for clinical use on the early diagnosis of hyperlipidemia (HL), neurologically and audiologically asymptomatic patients. METHOD: 30 random samples of HL patients who have normal blood pressure and no symptoms of circulatory system and nerve system were selected. As control group, another 30 men were selected. The clicks stimuli were presented at rates of 10/s and 50/s respectively in ABR test. RESULT: By the "ABR 10/s test" mode, the ABR showed no differences between two groups (P > 0.05). The net effect of ISR (50/s) on interpeak latency difference (IPLD) of ABR was significantly greater in the hyperlipidemic patients for IPLD(I-V) and IPLD(I III). In HL group, the effects were compared between retina angiosclerosis patients and no retina angiosclerosis patients and the same result was observed. CONCLUSION: ISR of ABR indicates a trend of subclinical impairments of brain stem function in hyperlipidemic patients, probably due to ischemia accelerated by their condition. PMID- 11944475 TI - [The thin sectional anatomy and MRI of the eustachian tube]. AB - OBJECTIVE: To provide the sectional anatomic data for clinical imaging diagnosis of the eustachian tube. METHOD: 9 cadaver eustachian tube were sectioned on transverse, coronal and sagittal planes using plastination, comparing with the MRI of the eustachian tube. RESULT: The coronal and transversal images clearly delineates the anatomical details of the eustachian tube and its fascia and muscles and their relationship, the fibrocartilaginous tube is posteromedial; its upper margin is curled outward to form the roof of the cartilaginous tube. Anteriorly and laterally, the eustachian tube is closed by the salpingopharyngeal fascia extending along the inferior surface of the eustachian tube to or its floor. The tensor veli palatini anterolateral and the levator veli palatini muscle posterolateral and inferior. The submucosal fascia is medial to the levator veli palatini muscle, the pharyngobasilar fascia is between the tensor and levator veli palatini muscles; the fascia of Weber Liel is lateral to the tensor veli palatini muscle. CONCLUSION: The cartilaginous portion, muscle and fascia of the eustachian tube could be displayed clearly on MRI. The thin sections of the eustachian tube can be directly compared with the MR images, providing valuable information to image diagnosis of the nasopharyngeal diseases. PMID- 11944476 TI - [Application of the scanning of three-dimensional space of whirl computed tomography in elongated styloid process]. AB - OBJECTIVE: Looking for a method of diagnosing elongated styloid process. METHOD: The scanning of three-dimensional space of whirl computed tomography (CT) has been used for 20 cases. RESULT: The patients who are suspected elongated styloid process are diagnosed really. CONCLUSION: The scanning of three-dimensional space of whirl CT is a very useful method for the diagnosing of suspected elongated styloid process. PMID- 11944477 TI - [The effects of BDP on calcium ion in the granular leukocytes in nasal secretions of the patients with allergic rhinitis]. AB - OBJECTIVE: To study the effects of beclomethasone dipropionate (BDP) on the inflammatory granular leukocytes in nasal secretions of the patients with allergic rhinitis. METHOD: The change of intercellular calcium ion of the eosinophils and neutrophils in the nasal secretions were observed under the laser scanning confocal microscopy and technology of fluorescence. RESULT: The concentration of intercellular calcium ion either in eosinophils or in neutrophils significantly rose after BDP treatment. CONCLUSION: BDP can rise the concentration of intercellular calcium ion in the granular leukocytes, which may be a factor of the anti-inflammation functions of BDP. PMID- 11944478 TI - [Outwardly rectifying potassium currents in isolated outer hair cells in cochlea of guinea pig]. AB - OBJECTIVE: The purpose of this study was to investigate electrophysiological properties of isolated outer hair cells (OHCs) from cochlea of the guinea pig, record outwardly rectifying potassium currents in OHCs with different length, analyze component of outwardly rectifying potassium currents, study kinetics of the currents. METHOD: The whole cell patch-clamp technique was used. RESULT: Whole-cell capacitance of the OHC was (30.96 +/- 2.79) pF(n = 29), zero current potential was (30 +/- 2.1) mV(n = 16) and reversal potential was (-51.67 +/- 1.84) mV(n = 9). Variation in expression of outwardly rectifying potassium currents was found in OHCs with different length, namely, potassium conductance was greater in short OHC than in tall OHC. The maximal amplitude of outwardly rectifying potassium currents were reduced by 60% in the presence of 100 mumol/L CdCl2 and change the kinetics of the currents, effects of CdCl2 on the peak currents were greater when compared to the effects on the steady-state currents (P < 0.01, n = 5). 1 mmol/L 4-aminopyridine (4-AP) inhibited 43% of the maximal amplitude of the currents and had no effect on the kinetics of the currents. Activation of the currents was described by Boltzmann equation, V1/2 = (-11.07 +/ 0.26) mV, S = (6.62 +/- 1.74) mV(n = 13). CONCLUSION: Outwardly rectifying potassium currents were composed of calcium-activated potassium current, outward delayed rectifier potassium current and A type current. PMID- 11944479 TI - [Electrically evoked auditory nerve compound action potentials in Nucleus CI24M cochlear implant users]. AB - OBJECTIVE: To study the feasibility and clinical applicability of electrically evoked auditory nerve compound action potentials (ECAP) in young patients with Nucleus CI24M cochlear implants. METHOD: ECAPs have been recorded from 37 Nucleus CI24M cochlear implant users in response to monopolar electrical stimulation of the cochlea by using the neural response telemetry(NRT) software and the hardware. Behavioral thresholds and maximum comfortable levels were measured in 12 of those patients. RESULT: ECAPs could be recorded in 87.6% of all patients. Good relations were observed between ECAP threshold and behavioral threshold, but poor relations between ECAP threshold and the impedance of different electrodes. We also observed growth functions and threshold variations of ECAP across subjects for a given place as well as across electrodes within a subject. These findings might be that these particular physiological measures are related to the number of living neural fibers and the excitability of the auditory nerve to electrical stimulation. CONCLUSION: ECAPs were proved to be useful in adjusting the stimulation parameters of the cochlear implant speech processor in order to maximize an individual's performance with the device, especially in young children. PMID- 11944480 TI - [The status of auditory function in auditory neuropathy]. AB - OBJECTIVE: To study the status of auditory function and the site of lesion in auditory neuropathy. METHOD: The data of 65 patients were analyzed including the clinic signs, pure tone audiometry, auditory brainstem response(ABR), 40 Hz auditory event related potential(40 Hz AERP) and otoacoustic emissions (OAEs). RESULT: The low frequency hearing loss in auditory neuropathy arose from the lesion of retrocochlear auditory afferent and efferent nerve and auditory brainstem, presented at the abnormality of acoustic reflex, efferent suppreSsion, ABR and the unparallel relationship between evoked OAE amplitude and pure tone threshold. To corresponding, the outer hair cells in low frequency region were out of suppression, presented at the strong response of SOAE, TEOAE and DPOAE in low frequency region. The high frequency hearing loss in auditory neuropathy arose from the lesion of cochlear outer hair cells, presented at the parallel relationship between DPOAE amplitude and pure tone threshold in high frequency region. The intermediate frequency hearing loss in auditory neuropathy was damaged gently or close to normal, presented at both pure tone threshold and DPOAE amplitude nearby 2 kHz were close to normal. CONCLUSION: The dysfunction in differ degree lied in the afferent and efferent nervous systems and cochlear level in auditory neuropathy, the main site of lesion was in the cochlear afferent and efferent nerve, up to the brainstem and down to the cochlea. PMID- 11944481 TI - [Patient counseling in the district hospital. Decreasing threshold anxiety]. PMID- 11944482 TI - [Diabetes counseling. The first meeting: foundation for success]. PMID- 11944483 TI - [Health promotion/prevention/managed care. Huge cost saving potential]. PMID- 11944484 TI - ["Together we are strong" (interview by Urs Luthi)]. PMID- 11944485 TI - [23rd Delegate Meeting. 1.5 million for the action fund]. PMID- 11944486 TI - [Legal principles of the strike. It means developing a strike culture]. PMID- 11944487 TI - [Call for global thinking and action]. PMID- 11944488 TI - [Council of national delegates. ICN opens its doors]. PMID- 11944489 TI - [A peaceful, melancholic mood]. PMID- 11944490 TI - [Self management promotes well-being in dialysis patients]. PMID- 11944491 TI - [Forum: the pen for you]. PMID- 11944492 TI - [The caring relationship--terminal care in neurosurgery. Between subject and object, the nomads of death]. PMID- 11944493 TI - [Nursing care in the 21st century. "Health care"--a utopia?]. PMID- 11944494 TI - [Venture forth from the silent chamber]. PMID- 11944495 TI - [Counseling principles. On the way to personal responsibility in managing illness]. PMID- 11944496 TI - [The "mobber": a victim himself?]. PMID- 11944497 TI - [H as in heroin]. PMID- 11944500 TI - Robert W. Morgan, DDS: U.S. Army Dental Corps, founder and creator of dental snuff. PMID- 11944501 TI - Is a dentist a physician? PMID- 11944502 TI - Orland lectureship. Heartland dentistry: a history of dental education and practice in Missouri and Kansas. PMID- 11944503 TI - Dentistry on stamps. PMID- 11944504 TI - Greene V. Black and "extension for prevention". AB - This study is an attempt to intervene in the contemporary debate on the interpretation of Greene V. Black's well known principle of "extension for prevention." It is possible to discern two, divergent strains within Black's thought: he contends that his principle should have universal application, at the same time as he seeks to limit its application. The conflict between these two positions may be resolved by arguing that Black's comments are directed at different audiences: Black does not actually believe that extension for prevention should be universally applied, but reserves the judgment not to extend to only the most experienced operators. The remainder, the majority of operators, should behave as if it were a universal principle. PMID- 11944505 TI - Lithium side-effects and predictors of hypothyroidism in patients with bipolar disorder: sex differences. AB - OBJECTIVE: To determine the prevalence of the side effects of lithium therapy and possible predictors of hypothyroidism in women and men with bipolar disorder. METHOD: Twenty-two men and 38 women with bipolar disorder and taking lithium for at least 1 year, were interviewed about lithium side effects using a list of the most commonly reported symptoms. RESULTS: The complaint most frequently reported was polyuria-polydipsia syndrome, which affected 36 (60%) of 60 patients. More men than women reported tremor (54% v. 26%, p < 0.05), but weight gain during the first year of treatment was more frequent in women than men (47% v. 18%, p < 0.05), as was the development of clinical hypothyroidism (37% v. 9%, p < 0.05). Weight gain during the first year of treatment (and not sex) was the only significant predictor of hypothyroidism. CONCLUSION: Weight gain during the first year of lithium treatment, in the absence of biological evidence of subclinical hypothyroidism, was the most predictive and, possibly, the first sign of hypothyroidism. PMID- 11944507 TI - Conversion sensory symptoms associated with parietal lobe infarct: case report, diagnostic issues and brain mechanisms. AB - This case report suggests that diagnostic difficulties and brain mechanisms related to conversion disorder associated with cerebral lesions differ from those related to conversion disorder without cerebral lesions. A 35-year-old divorced woman was admitted to a psychiatric inpatient unit with multiple physical complaints. The symptoms first appeared 5 years previous and 2 months after a sexual assault. Three years later, she began to experience ill-defined sensory symptoms confined to the left half of her body (splitting the midline). Results of neurologic consultations were equivocal because of the subjective nature of the complaints, which were viewed as conversion symptoms. A magnetic resonance imaging scan demonstrated an old infarct in the right parietal lobe, suggesting a physical origin of the patient's symptoms. However, normal somatosensory-evoked responses from the affected area contributed little to establishing the diagnosis. The results of all further investigations to identify causes of the vascular pathology were negative. The multiple ill-defined somatic symptoms, the distribution of sensory symptoms and the resolution of symptoms with psychotherapy established the diagnosis of conversion disorder superimposed on a pre-existing right parietal lesion. This case highlights the importance of clinical features in establishing a diagnosis such as this. We suggest that reactivation of implicit sensory memories (represented at the thalamic level and resulting from decreased corticofugal inhibitions due to the lesion) may contribute to the formation of sensory conversion symptoms in individuals with parietal lobe lesions. PMID- 11944506 TI - Cerebral activity associated with auditory verbal hallucinations: a functional magnetic resonance imaging case study. AB - Among the many theories that have been advanced to explain the mechanism by which auditory verbal hallucinations (AVH) arise, 2 that have received a degree of empirical support are: the hypothesis that AVHs arise from misinterpreted inner speech and the proposal that they arise from aberrant activation of the primary auditory cortex. To test these hypotheses, we were fortunate to be able to study the interesting and rare case of a woman with schizophrenia who experienced continuous AVH which disappeared when she listened to loud external speech. Functional magnetic resonance imaging (fMRI) was used to measure the patient's brain activity in the temporal and inferior frontal regions during the AVHs and while the she was listening to external speech. The brain activity of a matched control subject was also recorded under the same experimental conditions. AVHs were associated with increased metabolic activity in the left primary auditory cortex and the right middle temporal gyrus. Our results suggest a possible interaction between these areas during AVHs and also that the hypotheses of defective internal monitoring and aberrant activation are not mutually exclusive. Potential limitations to the generalization of our results are discussed. PMID- 11944508 TI - Rapid resolution of social anxiety disorder, selective mutism, and separation anxiety with paroxetine in an 8-year-old girl. PMID- 11944509 TI - Increasing sophistication of the pharmacotherapy of mood disorders. PMID- 11944510 TI - Assessment of premorbid function in first-episode schizophrenia: modifications to the Premorbid Adjustment Scale. AB - Studies have found Cannon-Spoor's Premorbid Adjustment Scale (PAS) to be a useful measure of premorbid function and an effective predictor of outcome in patients with chronic schizophrenia. Despite its widespread use, the applicability and reliability of the scale for use with young patients who experience their first episode of schizophrenia have not been thoroughly examined. We review the studies that used the PAS to assess premorbid function in patients with either chronic or first-episode schizophrenia. Difficulties that have been encountered with the use of the PAS in first-episode patients are presented, and modifications that have been made to the scale by various research groups are described. Finally, we make recommendations to enhance the use of the PAS when evaluating patients who have experienced their first episode of schizophrenia. PMID- 11944511 TI - [Primary clinical study on self-setting calcium phosphate cement in bone defect repair of extremities]. AB - OBJECTIVE: To investigate the clinical application of self-setting calcium phosphate cement (CPC) in bone defect repair of extremities. METHODS: From May 1998 to January 2000, 32 cases of bone defect, in 36 sites, were repaired and reviewed, aged from 4 to 59 years old (24.7 years old on average), with bone defect 2 to 125 cm2 in size (13.1 cm2 on average). The causes of the bone defect were fracture, bone cyst, iliac bone harvesting, fibrous dysplasia, enchondroma and bone tuberculosis, which involved femur, iliac, tibia, humerus, phalanx, fibula, calcaneus, talus and acetabulum. All of the cases were followed up for 1 to 23 months, 15.3 months on average, before radiographic examination. RESULTS: All operations were successful and no general response was observed in all of the cases. X-ray examination showed an integrity interface between CPC and bone. And CT showed no gap existed. There was no increase of serum calcium and phosphate levels. CONCLUSION: CPC is applicable in the low- or non-weight-bearing site of the extremities. PMID- 11944512 TI - [Application of prosthesis replacement in limb salvage treatment of mild malignant tumors at the articular ends of femurs]. AB - OBJECTIVE: To investigate the clinical outcome of application of joint prosthesis in limb salvage treatment of mild-malignant tumor at the articular ends of femurs, with severe osteolysis. METHODS: The treatment of 15 cases of mild malignant tumor at the articular ends of femurs with severe bone destruction, from 1978 to 1999, was reviewed. There were 10 cases of giant-cell bone tumor and 5 cases of chondrosarcoma, among which there were 5 cases at the stage of I A, 9 cases at I B, and 1 case at II A, with 4 cases at the proximal end and 11 cases at the distal end. The tumor was totally removed, with a massive bone defect left at the foci, and then the prosthesis replacement was performed to reconstruct the articular joint. All of the 15 patients were followed up for 9 months to 20 years, 4 years and 3 months on average, before clinical evaluation. RESULTS: All of the wound healed well, with primary healing. Local relapse occurred in one case and the patient died of lung metastasis; the amputation of the replaced knee joint was performed in one case due to severe infection after trauma; radiograph showed there was slight sunk of the prosthesis in 2 cases after replacement. The other 11 cases recovered well with satisfactory function. CONCLUSION: The limb salvage treatment of mild-malignant tumor at the articular ends of femurs would be available, especially for those with massive bone destruction, when the tumor is removed by whole and rationally marginal resection, followed by properly utilization of prosthesis and general post-operative exercise. PMID- 11944513 TI - [Surgical approach in plastic of shortened lower limb]. AB - OBJECTIVE: To introduce a new surgical approach to rectify the shortened lower limbs. METHODS: From March 1985 to October 2000, 288 cases of shortened lower limbs were treated and reviewed. Closed fracture at the metaphysis was made by a self-made "needle saw", and then the "multiple-plane and double-track elongation instrument" was adopted to elongate the fractured bone. There were totally 161 cases of male and 127 cases of female included, with average age 21.3 years old, ranging from 12 to 29 years old, among which there were 268 cases elongated at the proximal metaphysis of the tibia, 16 cases at the distal femur and 4 cases at the distal tibia. All of the cases were followed up for 6 to 8 months before clinical evaluation. RESULTS: The lower limbs in 288 cases were elongated for 3.0 to 11.5 cm in 24 to 96 days, averaging 47 days, which fulfilled pre-operative plan. In the second week after the operation, new calculus and periosteum formed obviously in the gap between the fractured parts, and in 6 to 8 months bone union was observed at the fractured site in all cases. There was no nerve or blood vessel injury, or non-union of the metaphysis fracture. The function of the manipulated knee joints and ankle joints recovered well. CONCLUSION: It is a practical and safe surgical option to rectify the shortened lower limbs by closed fracture at the metaphysis, followed by elongation of the fractured bone, without any complication such as non-union or atrophy of manipulated bone, and with no need of internal fixation or bone grafting. PMID- 11944514 TI - [Comparison of long bone repair in tibia by vascularized fibular grafting of different sides]. AB - OBJECTIVE: To evaluate the clinical effect of repair of massive bone defect in tibia by vascularized fibula grafting of either sides. METHODS: Twenty-four cases of massive bone defect in tibia, among which 14 case were repaired by vascularized fibula grafting of the other side and another 10 cases were repaired by those of the same side, from 1987 to 1997 were followed up for 3 to 13 years; the functions of the operated limbs were evaluated according to Enneking Score System, and the outcome of the fibula grafts were assessed by radiographic examination with reference to the standard established by International Symposium on Limb Salvage. RESULTS: The average recover rate of the operated limbs in those repaired by the other side grafting was 80.7%, and the average healing period of the fibula graft was 14 weeks with fracture of the graft in one case which made the operated lower limb shorten for about 2.5 cm; the fibula grafts were observed thickened in 43 weeks, on average, and the patients could walk independently without a crutch. While in those repaired by the same side grafting, the average recover rate of the operated limbs was 68.3%, the average healing period of the fibula graft was 17 weeks with fracture of the graft in 3 cases, in 2 of which the lower limbs were shortened for 2 cm and 4 cm respectively, and in the third one infection occurred and amputation was performed finally; the fibula grafts were observed thickened in 49 weeks, on average, which made it available for the patients to walk without a crutch. All of the data showed that there was a significant difference statistically between the differently treated cases. CONCLUSION: It's a good choice to repair massive bone defect in tibia by vascularized fibula grafting, and the vascularized fibula graft from the other side could promote the bone healing and accelerate the recover of the function of the operated lower limb. PMID- 11944515 TI - [Experimental study on stimulation of guided bone regeneration by autogenous bone marrow]. AB - OBJECTIVE: To study the effect of autogenous bone marrow on guided bone regeneration (GBR), and evaluate the repairing ability of GBR in bone defect with autogenous bone marrow. METHODS: Ten mm segmental defects were produced in both radii of 18 rabbits. The defect was bridged with a silicon tube. Autogenous bone marrow was injected into the tube on the experimental group at 0, 2, 4 weeks after operation, and peripheral blood into the control group at the same time. The X-ray, gross, histological and biochemical examinations were observed in various times. RESULTS: The new bone formation of experimental group was prior to that of control group; calcium and alkaline phosphatase of experimental group were higher than those of control group. The experimental group had all been healed at the tenth week, but no one healed in control group. CONCLUSION: It can be conclude that autogenous bone marrow can stimulate bone formation and facilitate GBR in bone defect. PMID- 11944516 TI - [Primary study on repair of mandibular defect using free fibular composite flap]. AB - OBJECTIVE: To summary the clinical application of free fibular composite flaps in repair of mandibular defect. METHODS: Fifty-eight cases of mandibular reconstruction using free fibular flaps, from June 1999 to November 2000, were reviewed, among which there were 37 cases of male and 21 cases of female, aged from 12 to 65 years old. All of the relevant data of the operation, such as the cause of the defect, design of the flaps, the blood vessels of the recipient and the complications, were included and analyzed. RESULTS: The fibular flap was 10.9 cm (4-21 cm) averagely in length and was harvested for 2.1 times (1-4 times); the average size of the skin paddle was 5.88 cm x 3.13 cm. Of 58 flaps, there were 56 survived; the overall complication rate was 24.1%. CONCLUSION: It's safe and reliable to use free fibular flap to reconstruct the mandibular defect. PMID- 11944517 TI - [Transferring of the pedicled second metatarsal base for repairing bone defect of lateral malleolus]. AB - OBJECTIVE: To study the method and effect of transferring the pedicled second metatarsal base for repairing bone defect of lateral malleolus. METHODS: Thirty lower limb specimens were anatomized to observe the morphology, structure and blood supply of the second metatarsal bone. Then transferring of the pedicled second metatarsal base was designed and used in 6 patients clinically. All cases were male, aged from 24 to 48 years old, and the area of bone defect was 3-4 cm. RESULTS: Followed up for 3-11 months, all patients healed primarily both in donor and recipient sites. There were excellent results in 4 cases and good results in 2 cases. The morphology and function of the malleoli were satisfactory. CONCLUSION: Transferring of the pedicled second metatarsal base for repairing bone defect of lateral malleolus is an effective and reliable operative method. PMID- 11944518 TI - [Surgical treatment of lumbar spondylolisthesis with cages and transpedical screws]. AB - OBJECTIVE: To observe the effect of threaded titanium cage and transpedical screw for the treatment of lumbar spondylolisthesis. METHODS: Eighteen patients with lumbar spondylolisthesis were adopted in this study. Among them, there were 8 males and 10 females, aged from 43 to 62 years old. Roentgenogram showed that there were 6 cases of I degree spondylolisthesis, 11 cases of II degree and 1 case of III degree. All patients were treated with cages for intervertebral fusion after total laminectomy and pedicle screws for the reduction. RESULTS: The cases were followed up from 6 to 12 months with an average of 11 months. The clinical results were excellent in 13 cases and good in 5 cases. All patients achieved successful fusion and bony union. There were no pedicle screw loosening or broken or peripheral nerve dysfunction in this series. CONCLUSION: This method has been proved to be an effective and reliable procedure for treatment of lumbar spondylolisthesis. It produces a high fusion rate and clinical success. PMID- 11944519 TI - [Locking compression plate fixation for periprosthetic femoral fracture]. AB - OBJECTIVE: To introduce a method for fixation in periprosthetic fracture with locking compression plate (LCP). METHODS: During the surgery, a long 12-hole LCP was placed to the lateral side of the femur. Six holes of the plate were placed proximal to the main fracture line to make sure that there could be enough cortex units for rigid fixation of proximal fragments. Locking screws used except for the most proximal hole where a 4.5 mm screw was used instead. RESULTS: The patient was pain free at the fracture site one week after the surgery, and was able to walk with a cane 3 months after the surgery. Bone union was evident radiographically 3 months after the surgery. There was no loose signs around the stem. Six months after the surgery, the patient recovered full function without pain. CONCLUSION: In our experience from this case, LCP in treating periprosthetic fracture was easy and less time consuming, more over, extensive periosteal stripping could be avoided. LCP is a good choice in treating some periprosthetic fractures. PMID- 11944520 TI - [Transforming growth factor-beta superfamily in the regulation of new bone formation]. AB - OBJECTIVE: To review the recent advances in transforming growth factor-beta(TGF beta) superfamily study and its role in new bone formation. METHODS: The latest original articles related to this subject were retrieved extensively, especially the effect of TGF-beta, bone morphogenetic proteins(BMPs) and activin(ACT) on distraction osteogenesis. RESULTS: TGF-beta, BMPs and ACT play important roles in prompting new bone formation and each of them has different effects. Among them, TGF-beta can stimulate the proliferation of osteoblast and synthesis of extra cellular medium; BMPs can initiate the differentiation of interstitial cell to osteocyte; then ACT displays the combine effect of above two factors. CONCLUSION: TGF-beta superfamily can regulate new bone formation and thus shorten the course of mandibular distraction osteogenesis. PMID- 11944521 TI - [Tissue engineering study on chitosan-gelatin/hydroxyapatite composite scaffolds- osteoblasts culture]. AB - OBJECTIVE: To investigate the behavior of rat calvarial osteoblasts cultured on chitosan-gelatin/hydroxyapatite (CS-Gel/HA) composite scaffolds. METHODS: The rat calvarial osteoblasts (the 3rd passage) were seeded at a density of 1.01 x 10(6) cells/ml onto the CS-Gel/HA composite scaffolds having porosity 85.20%, 90.40% and 95.80%. Cell number was counted after cultured for 3 days, 1 week, 2 weeks and 3 weeks. Cell proliferation, bone-like tissue formation, and mineralization were separately detected by HE, von Kossa histological staining techniques. RESULTS: The CS-Gel/HA composite scaffolds supported the attachment of seeded rat calvarial osteoblasts. Cells proliferated faster in scaffold with higher porosity 90.40% and 95.80% than scaffold with lower porosity 85.20%. The osteoblasts/scaffold constructs were feasible for mineral deposition, and bone like tissue formation in 3 weeks. CONCLUSION: This study suggests the feasibility of using CS-Gel/HA composite scaffolds for bone tissue engineering. PMID- 11944522 TI - [Influence of dosage on cell biocompatibility of hydroxyapatite/tricalcium phosphate]. AB - OBJECTIVE: To investigate the influence of different dose levels of hydroxyapatite/tricalcium phosphate (HA/TCP) on the proliferation and alkaline phosphatase (ALP) activity of rabbit osteoblasts. METHODS: Three different dose levels of HA/TCP (10%, 40%, 70%) were co-cultivated with rabbit osteoblasts respectively. The proliferation and ALP expression capacity of osteoblasts were examined with MTT method and enzyme histochemistry once every 24 hours until 5 days. Three control groups of other materials were treated and examined in the same way: rabbit osteoblasts as normal control; polyvinylchloride as positive control; titanium alloy as negative control. RESULTS: There was remarkable time effect relationship in the proliferation of osteoblasts. Ten percent HA/TCP did not affect osteoblasts growth while 40% HA/TCP could slow the cell growth rate down though time-effect relationship still existed. The proliferation of osteoblasts stagnated when co-cultivated with 70% HA/TCP. On the other hand, 10% HA/TCP could cause reversible damage on ALP activity of osteoblasts, whereas when the dose was 40%, and the cultivation lasted 6 days the damage was irreversible. Three different dose levels of titanium alloy (10%, 40%, 70%) had no effect on the proliferation or ALP activity of osteoblasts. CONCLUSION: Dosage is an important factor affecting the biocompatibility evaluation of biomaterial. It suggests that dose choosing should be more specified upon each individual biomaterial. It also indicates that ALP may be a good supplementary index of the cell compatibility of material. PMID- 11944523 TI - [Peripheral nerve regeneration under immunosuppression]. AB - OBJECTIVE: To discuss peripheral nerve regeneration under immunosuppression. METHODS: Current research trends about relationship between peripheral nerve injury and immunoreaction, the experimental result of nerve regeneration after using various immunosuppressors, and the clinical findings after human allogenous hand transplantation were extensively reviewed. RESULTS: Peripheral nerve regeneration was accelerated under immunosuppression. CONCLUSION: Peripheral nerve injury may induce immunoreaction, which inhibit nerve regeneration and function recovery. PMID- 11944524 TI - [Effect of vascular endothelial growth factor in fracture healing]. AB - OBJECTIVE: To observe the effect of vascular endothelial growth factor (VEGF) in fracture healing and to investigate the influence of VEGF and VEGF antibody in fracture healing. METHODS: One hundred and five rabbits were used to make fracture model in the left radius and randomly divided into control group, VEGF group and VEGF antibody group. VEGF and VEGF antibody were used in the VEGF group and VEGF antibody group respectively, then the blood flow of the fracture ends was measured by single photon emission computed tomography (SPECT) 8, 24, 72 hours, 1, 3, 5 and 8 weeks after fracture, the X-ray films of the fracture sites were taken after 1, 3, 5 and 8 weeks to observe the fracture healing. RESULTS: The blood flow of the fracture ends in the VEGF group increased during a period from 8 h to 3 wk after fraction when compared with that of the control group, and no obvious difference was seen on the X-ray films between the two groups. In the VEGF antibody group, the blood flow of the fracture ends decreased obviously when compared with that of the control group. The fracture healing process was interfered seriously and nonunion change was seen in the fracture site. CONCLUSION: The lack of VEGF will interfere with the fracture healing process and result in nonunion in the fracture site. Administration of ectogenous VEGF may promote fracture healing by increasing the blood flow of the fracture ends. PMID- 11944525 TI - [Preparation of rhBMP-2/BCB reconstituted bone xenograft and assay of its osteoinductivity]. AB - OBJECTIVE: To investigate a new grafting material of bone xenograft with strong bone inductive and conductive capacity. METHODS: Based on successful clinical application of the reconstituted bone xenograft (RBX), a new xenograft was made by combining recombinant human bone morphogenetic protein-2 (rhBMP-2) with antigen-free bovine cancellous bone (BCB). Sixty male BALB/C mice aged 4 weeks were divided into study group of 30 and control group of 30 randomly. rhBMP-2/BCB was implanted in the left thigh muscle pouch in the study group and BCB in the control group. The mice were sacrificed at 7 d, 14 d and 21 d after implantation. Inductivity of rhBMP-2/BCB was detected by histological observation and biochemical determination of the samples. RESULTS: Histological examination showed that rhBMP-2/BCB induced chondrogenesis on the 7th day, with woven bone formed on the 14th day, and lamellar bone and marrow on the 21st day, while BCB failed to induce chondrogenesis or osteogenesis on the 7th, 14th and 21st days. The alkaline phosphatase activities and calcium content in study group were higher than those in control group with significant difference (P < 0.01). CONCLUSION: rhBMP-2/BCB is an ideal grafting material with strong bone inductive and conductive capacity without evoking immune reaction. PMID- 11944526 TI - [Mode of new bone formation following distraction osteogenesis in mandibular lengthening]. AB - OBJECTIVE: To investigate the mode and influential factor of new bone formation following distraction osteogenesis in mandibular lengthening. METHODS: Corticotomy was performed on bilateral mandibles in twelve adult male goats. A custom-made distractor was used to lengthen the mandible at a rate of 1 mm/day for 10 days (total 10 mm elongation). Four goats were sampled respectively at 2, 4 and 8 weeks after completion of distraction. The lengthening mandibles were examined by roentgenography and histology. RESULTS: Newly formed callus was observed in the distraction gap after mandibular lengthening. The new bone exhibited intramembranous ossification generally, but cartilage islands could be found in the specimen that distractor loosed. CONCLUSION: The above findings indicate that the mode of new bone formation in mandibular lengthening following distraction osteogenesis appears to be intramembranous ossification and that endochondral ossification takes place in case distractor has loosened. PMID- 11944527 TI - [Study of rat osteoblasts transfected by transforming growth factor-beta 1 gene]. AB - OBJECTIVE: To investigate the effect of transforming growth factor-beta 1 (TGF beta 1) gene transfer on the biological characteristics of osteoblasts. METHODS: The expression of TGF-beta 1 in the transfected osteoblasts was detected by in situ hybridization and assay of TGF-beta 1 activity in the supernatant (mink lung epithelium cell growth-inhibition test). The effects of gene transfer and supernatant of the transfected osteoblasts on the proliferation and alkaline phosphatase(ALP) activity of osteoblasts were detected by 3H-TdR and MTT. RESULTS: The results of in situ hybridization analysis suggested that the osteoblasts transfected by TGF-beta 1 gene could express TGF-beta 1 obviously. The complex medium, which was the mixture of serum-free DMEM and the activated supernatant according to 1:1, 1:2, 1:4, could inhibit growth of Mv-1-Lu evidently and the ratios of inhibition were 16.3%, 22.7%, 28.2% respectively. TGF-beta 1 gene transfer had no effect on the biological characteristics of osteoblasts, but the activated supernatant of transfected osteoblasts stimulated proliferation and inhibited ALP activity of osteoblasts. CONCLUSION: TGF-beta 1 gene transfer promotes the expression of TGF-beta 1 and the biological characteristics of transfected osteoblasts are stable, which is helpful for gene therapy of bone defects in vivo. PMID- 11944528 TI - [Role of recombinant human bone morphogenetic protein 2/collagen as an onlay bone graft on adult rat calvarial bone]. AB - OBJECTIVE: To evaluate the bone regenerative potential of recombinant human bone morphogenetic protein 2(rhBMP-2)/collagen on adult rat calvarial bone. METHODS: A tight subperiosteal pocket was produced under both sides of the temporal muscle in rats. rhBMP-2/collagen was implanted in one side and collagen alone was implanted in the other side as control. The rats were sacrificed 2, 4 and 8 weeks after operation. The specimen was harvested and examined histologically. For morphometric analysis, the thickness of the temporal bone of both sides was measured and compared. RESULTS: The rhBMP-2/collagen onlay implant resulted in active bone formation and the augmented bone was connected directly with the original bone, whereas the collagen alone resulted in neither bone nor cartilage production. The ossification process in the rhBMP-2/collagen occurred directly through bone formation, similar to intramembranous ossification. CONCLUSION: rhBMP-2/collagen is an effective material as a biological onlay implant. PMID- 11944529 TI - [Experimental study on repair of bone defect in femoral head by enhanced autogenous bone combined with bone morphogenetic protein]. AB - OBJECTIVE: To compare and evaluate the capability of pure autogenous bone and the enhanced autogenous bone combined with bone morphogenetic protein in bone repair of femoral head. METHODS: Eighteen femoral heads of 9 dogs were drilled by trephine, 4 mm in diameter, followed by respective implantations of autogenous bone grafting (group B) and of the enhanced autogenous bone composite, combined with bone morphogenetic protein (group C), with the self-repair of bone defect as the control (group A). Three, six, nine weeks after the operation, radiological examination, computerized tomography, light and electronic microscopes were performed to investigate the bone healing of the defect in the femoral head. RESULTS: In group A, it could be observed that there was hematoma organization and delayed woven bone formation in the 3rd week after operation, and there was little replacement of woven bone by bone trabecula in the 9th week; in group B, the autogenous bone implanted were dead in the 3rd week and maintained in situ in the 9th week; in group C, active new bone formation, either endochondral or intramembranous ossification, was found in the 3rd week and entire repair of the bone defect by bone trabecula in the 9th week after operation. CONCLUSION: The enhanced autogenous bone combined with bone morphogenetic protein could promote reconstruction of the bone defect in femoral head, superior to pure autogenous bone which could provide a framework for the new bone formation. PMID- 11944530 TI - [Biomechanical study on the composite of allogenic decalcified bone matrix gelatin and bone cement]. AB - OBJECTIVE: To evaluate the biomechanical properties and structural characteristics of various composites of partially decalcified allogenic bone matrix gelatin and bone cement at different ratios. METHODS: According to Urist method, partially decalcified allogenic bone matrix gelatin was prepared and mixed with bone cement at different ratios of 0, 400, 500, and 600 mg/g. Then the comparisons of these composites were performed in microstructure, ultimate compression strength and ultimate bending strength properties. RESULTS: The electronic microscope showed that the bone particles and bone cement were distributed evenly in the composite, irregularly connecting by multiple points; with the increase of bone particles and decrease of bone cement in the composite, there were more and more natural crevices, varying from 100 microns to 400 microns in width, in the biomaterials. Of all the composites with the ratios of 0, 400, 500, and 600 mg/g, the measurements of ultimate compression strength were (71.7 +/- 2.0) MPa, (46.9 +/- 3.3) MPa, (39.8 +/- 4.1) MPa, and (32.2 +/- 3.4) MPa, respectively; and the measurements of ultimate bending strength were (65.0 +/- 3.4) MPa, (38.2 +/- 4.0) MPa, (33.1 +/- 4.3) MPa and (25.3 +/- 4.6) MPa, respectively. CONCLUSION: The composite of partially decalcified allogenic bone matrix gelatin and bone cement has a good biomechanical property and could be easily fabricated and re-shaped, which make it available to be used clinically as an idea bone graft biomaterial. PMID- 11944531 TI - Evidence of our instincts: an interview with Linda H. Aiken. Interview by Alison P. Smith. PMID- 11944532 TI - Quality indicators for academic nursing primary care centers. AB - ANCs, by definition, deliver more than clinical services to patients and communities. The unique identifier that separates ANCs from other primary care/ambulatory care centers is the educational service they offer to students and other faculty in the school of nursing to which they belong. Therefore, measuring the quality of an ANC must include a measurement of the educational properties that it possesses in addition to the usual quality measurements by such organizations as AAAHC, JCAHO, CHAP, and AAACN. Unless these properties are included and measured by an ANC, it becomes difficult to justify the existence of such a clinic within a school of nursing. ANCs are encouraged to develop quality evaluation programs aimed at evaluating the educational aspects as well as the administrative and clinical aspects of their operations. PMID- 11944533 TI - Turnover of licensed nurses in skilled nursing facilities. PMID- 11944534 TI - Worker-designed tools: developing comprehensive work assessment tools and competencies. PMID- 11944535 TI - Warming your heart: the energy solution. AB - Citron (2002) probably sums it up best when he states that generosity is the best strategy for corporate prosperity. He believes that the love you get from others as a person and a corporation is equal to the love you extend to others. It's really about creating communities of caring and missions of inclusiveness. And the way to make this all happen is to continually recharge our batteries by staying close to the beautiful stories of our organization that will undeniably warm our hearts and give us the energy to be generous. PMID- 11944536 TI - Computerized physician order entry: a prescription for patient safety. AB - Clinician adoption of CPOE/CDS solutions is crucial to helping caregivers reduce medical errors and enhance patient safety. The LeapFrog Group CPOE/CDS report can be a helpful guide, but as clinicians concerned about the quality of health care and the well-being of our patients, we must play an active role in the successful adoption of these solutions by: 1. Making sure that your institution is committed to having the appropriate people involved in the entire process, including nurse leaders. 2. Selecting a vendor that has the knowledge and clearly understands the importance of implementing this type of system. 3. Ensuring that your organization is selecting a system that actually meets the criteria defined by the LeapFrog Group. PMID- 11944537 TI - Grassroots recruiting. PMID- 11944538 TI - Safety technology: solutions or experiments? PMID- 11944539 TI - Ambulatory care nursing practice: developing and contributing to the evidence base. PMID- 11944541 TI - Nursing shortage: Johnson & Johnson campaign aims to increase awareness, generate interest. PMID- 11944540 TI - HIPAA: update on rule revisions and compliance requirements. AB - Due to the highly technical requirements for HIPAA compliance and the numerous administrative and clinical functions and processes involved, guidance from experts who are knowledgeable about systems design and use to secure private data is necessary. In health care organizations, this will require individuals who are knowledgeable about clinical processes and those who understand health information technology, security, and privacy to work together to establish an entity's compliance plans and revise operations and practices accordingly. As a precondition of designing such systems, it is essential that covered entities understand the HIPAA's statutory requirements and timeline for compliance. An organization's success in preparing for HIPAA will depend upon an active program of assessment, planning, and implementation. Compliance with security and privacy standards can be expected to increase costs initially. However, greater use of EDI is expected to reduce costs and enhance revenues in the long run if processes and systems are improved. NOTE: Special protection for psychotherapy notes holds them to a higher standard of protection. Notes used only by a psychotherapist are not intended to be shared with anyone and are not considered part of the medical record. PMID- 11944542 TI - [Teenagers and the "morning after pill"]. PMID- 11944543 TI - [Infectious stomatitis]. AB - Infectious stomatitis is bacterial essentially when of dental origin, the viral forms most often causing a vesicular and erosive stomatitis, and fungal secondary to a modification of the oral commensal flora. The diagnosis is often clinical with the lesions being of characteristic appearance and arising in a suggestive context. When the appearance is less typical or when the choice of treatment necessitates the identification of the germ, specific samples are taken. The treatment of viral stomatitis is essentially symptomatic, that of bacterial and fungal stomatitis comprising two arms: the specific treatment of the acute episode and then that of the particular oral-dental terrain. PMID- 11944544 TI - [Cheilitis]. AB - Cheilitis is clinically characterised by an acute or chronic inflammatory state of the lips, limited to the vermillion border. The term "angular" cheilitis applies to localise bilateral lesions of the angles of the mouth. Acute cheilitis is erythematous or erosive, accompanied by a stinging sensation. Chronic cheilitis is for the most part erythemato-squamous, and is often asymptomatic. The classification of cheilitis is based on the aetiology, being most often an allergy, an infection or a physical irritation, potentially favourised by particular anatomical conditions. However, the distinction between the diverse types is sometimes blurred as diverse causes can co-exist. PMID- 11944545 TI - [Diagnosis of mouth ulcers]. AB - Oral ulcerations have very varied characteristics, the analysis of which is often sufficient for the clinician to establish a diagnosis and propose a suitable treatment. Aphthous ulcers and traumatic lesions are the most frequent, however one also sees infectious ulcerations (tuberculous, syphilitic, common germs or viruses), inflammatory diseases of the gastrointestinal tract, bullous dermatosis of autoimmune, toxic or unknown origin such as erythema multiform, and erosive lichen planus. It is thus the general context or the evolution that orientates the diagnosis, which is made more precise in certain cases by complementary investigations, in particular by the biopsy. When the ulceration is isolated it is necessary to first exclude the much-feared squamous cell carcinoma, but also other malignant ulcerated tumours, lymphomas, and tumours of the salivary glands. Thus it is imperative that the histological examination confirms the diagnosis, to formulate a prognosis and to allow the commencement of treatment. PMID- 11944546 TI - [Aphthous stomatitis]. AB - Aphthous stomatitis remains an illness of unknown aetiology, probably drawing on immunological mechanisms. The diagnosis of an oral aphthous ulcer is clinical, corresponding to a precise primary lesion. The benign form is the most frequent, however the illness sometimes evolves into a severe form which can then be considered as a generalised illness. The management of the patient must distinguish the evolving clinical form of the aphthous ulcer, search for a likely aetiology, and employ a therapeutic approach, which actually remains disappointing with the basic treatments being of solely temporary efficacity. PMID- 11944547 TI - [Bullous diseases of the oral mucosa]. AB - Bullous diseases of the oral cavity cause painful, erosive lesions. These must be distinguished from aphthous ulcers and vesicles with which they are often confused. The causes of post-bullous erosions are varied. Acute lesions can be due to trauma, erythema multiforme or toxidermia. The examination of the skin and the medical history are the keys to diagnosis. Chronic bullous diseases can be due to congenital diseases (epidermolysis bullosa or lymphangioma), which are easily evoked by the age at which the symptoms appear and the clinical picture. Acquired chronic bullous diseases include lichen planus and bullous auto-immune diseases. The keys to diagnosis are the presence of other mucosal or cutaneous lesions, the histological examination and the finding of perilesional antibodies on the mucosa. The symptomatic treatment of oral bullous lesions includes the following of a specific diet, use of antalegics and the prevention or treatment of superimposed infection. PMID- 11944548 TI - [White lesions of the oral mucosa]. AB - In this review, the etiologies of white oral lesions will be discussed. Thrush is the commonest form of oral candidiasis and it is the prototype of pseudomembranous lesions. The latter are whitish lesions that can be easily rubbed off. All patients with erosive or ulcerative lesions of the oral mucosa, e.g. patients with immunobullous diseases, can present with such lesions. Leucokeratosis refers to white, firmly adherent, lesions. A great number of diseases can manifest with such lesions: inflammatory diseases like lichen planus, infectious diseases like chronic candidiasis or hereditary diseases like the white sponge nevus. Leucoplakia refers to keratosic lesions tobacco-induced or that can not be precisely classified clinically. Such lesions are considered premalignant. When other clinical signs like erythema or ulceration are associated, these lesions are often epidermoid carcinomas. PMID- 11944549 TI - [Iatrogenic stomatitis]. AB - The oral cavity is the site of diverse manifestations of iatrogenic disease. Mucosal pigmentations, gingival hypertrophy, ulcerations, mucositis and xerostomia are easily identifiable, but may simulate diseases that are well known to localise to the oral cavity, such as aphthous ulcers or lichen planus. Drug therapies are at the origin of the greatest number of these lesions. A rigorous inquiry into the characteristic oral lesions permits, in the majority of cases, the establishment of this relationship. PMID- 11944550 TI - [Oral pain]. AB - Pain, a major symptom of stomatological disease, usually leads to a specialist consultation. Most commonly it is caused by dental caries and differs in nature and in intensity according to the stage of disease: dentinitis, pulpitis, desmodontitis and dental abscess. Added to this is peridental pain and the pre- and post-operative pains related to these diseases. Almost all oral-maxillary pathology is painful, be it boney such as in osteomyelitis and fractures, mucosal in gingivo-stomatitis and aphthous ulcers, or tumourous. However, besides the "multidisciplinary" facial pains such as facial neuralgia and vascular pain, two pain syndromes are specific to stomatology: pain of the tempero-mandibular joint associated with problems of the bite and glossodynia, a very common somatic expression of psychological problems. PMID- 11944551 TI - [A century of women in medicine]. PMID- 11944552 TI - [Pneumothorax]. PMID- 11944553 TI - [Superficial adenopathy]. PMID- 11944554 TI - [Adult hypothyroidism]. PMID- 11944555 TI - [Gallstones and their complications]. PMID- 11944556 TI - [Acute and chronic headache]. PMID- 11944557 TI - [Chronic venous insufficiency. Varicose veins]. PMID- 11944558 TI - [Induced abortion]. PMID- 11944559 TI - A reasonable approach to physical security. PMID- 11944560 TI - Climbing the coding career progression ladder. PMID- 11944561 TI - How trends shape the work force today and tomorrow. AB - What forces are propelling the healthcare industry's prolonged period of change? How are these forces shaping the future of HIM? In this article, investigators from the Center for Health Workforce Studies examine the roots of the work force shortage and reveal trends that will continue to influence the industry. PMID- 11944562 TI - A recruitment partnership for the future. PMID- 11944564 TI - Leaping ahead with patient safety. PMID- 11944565 TI - Megatrends promise transcription changes. PMID- 11944566 TI - Practice brief. HIPAA privacy and security training. PMID- 11944567 TI - The changing face of public health reporting. PMID- 11944568 TI - ICD-9 committee explores new technology, drug codes. PMID- 11944569 TI - Coding on the home front. AB - Recently, a group of coders participated in a survey about their day-to-day experiences as remote, or at-home coders. The survey asked a series of questions related to the positives and negatives of coding at home versus coding in the more traditional office setting. Below is a summary of the survey findings. PMID- 11944570 TI - Committed to connectivity. MedUnite creates an ATM-like network for thousands of healthcare providers and health plans. PMID- 11944571 TI - A chance to heal. Home health agencies can improve patient care and increase profits with telehealth wound consulting. PMID- 11944572 TI - Tracking a decade of CIS (clinical information systems). A vendor survey tracks 10 years of CIS use in the obstetrics department of a nationally renowned medical center with some stunning results. PMID- 11944573 TI - High-tech, low-cost, huge results. Health plan employs a technology-driven DM program that delivers big benefits for participating members. PMID- 11944574 TI - Patient-centric HIS. A healthcare information system based on a longitudinal patient record provides benefits to patients--and clinicians, administrators and IT staff as well. PMID- 11944575 TI - What works: claims coding/processing, putting claims on the right path. Medical care provider replaces manual processing system with an automated claims management sysytem and reduces turnaround by more than 50 percent. PMID- 11944576 TI - Managed care hotlist. PMID- 11944577 TI - Open technologies and the quality of healthcare delivery. PMID- 11944578 TI - Familial clustering of end-stage renal disease in Mississippi. AB - BACKGROUND: Familial clustering of ESRD has been noted in the families of 20-40% of African-Americans (AA) with ESRD but has been observed much less often in Caucasians (Cau). Because our impression indicated a higher frequency in Caucasians than reported in several recent studies we conducted a 2-year prospective evaluation of patients referred for renal transplantation to assess the frequency of familial clustering of ESRD in both Caucasians and African Americans. METHODS: Detailed family histories were obtained in 350 ESRD patients referred for renal transplantation, 335 of whom could provide appropriate information regarding first- and second-degree family members (252AA and 83 Cau) who were maintained on out-patient dialysis at least one month or had received a renal transplant. RESULTS: Patient race and sex paralleled that of the current Mississippi ESRD population but mean age was 17 years younger in the transplant candidates (39.9 vs. 57.2 yrs). If polycystic kidney disease (PKD) is included in the analysis, 31.7% of AA and 30.1% of Cau had at least one other family member with ESRD. If PKD is eliminated then 30.9% of AA and 25% of Cau had at least one other family member with ESRD. In both races the frequency of a positive family history was greatest in index cases with chronic glomerulonephritis, and of those with renal biopsies (n = 59), patients with focal segmental glomerulosclerosis (FSGS) and systemic lupus erythematosus (SLE) had the strongest family histories of ESRD (40% and 36%, respectively). Cau and AA with FSGS had comparable familial clustering of ESRD (41% vs, 38%) but SLE was confined to AA. CONCLUSIONS: In younger individuals with ESRD, familial clustering of end stage renal disease occurs with almost equal frequency in Caucasians and in African Americans. Screening of family members of index cases in both races might define a high-risk group of patients in whom prophylactic measures might be directed to thwart the progression [table: see text] of renal failure. PMID- 11944579 TI - Looking back: a blood test for the determination of sex in utero by means of dialysis in 1915. PMID- 11944580 TI - Estrogen receptors in canine mammary gland tumours. AB - The aim of the present study was to establish the relationship between estrogen receptor expression in neoplastic parenchymal cells and the type of canine mammary gland tumour. The research material included mammary gland tumours obtained from 66 dogs. Paraffin sections were stained with hematoxylin-eosin and immunohistochemically with monoclonal antibodies and LSAB/Peroxidase/Universal Kit. The estrogen receptor expression was observed in nuclei of neoplastic cells in about 59% of the cases and in the cytoplasm in about 89% of the cases. In about 20% of the cases the expression in the nuclei and in the cytoplasm was extremely weak. No correlation was found between the expression of estrogen receptors and the value of mitotic indexes in the neoplasms investigated. PMID- 11944581 TI - Hormonal profile and morphological changes in pig ovaries after intraovarian infusions of Escherichia coli endotoxin. AB - The purpose of this study was to estimate morphological changes in the ovary and size of the production of steroid hormones during the luteal phase of the estrous cycle in pigs after intraovarian infusions of Escherichia coli endotoxin. Polish Large White gilts (n = 15) of similar age (7-8 months) and body weight (90-110 kg) with two controlled subsequent estrous cycles were used. The animals were randomly divided into two groups: control (n = 9, the 10th day of the estrous cycle,) and treated with Escherichia coli endotoxin (n = 6, the same day of the estrous cycle). The gilts were infused with Escherichia coli endotoxin at a dose of 1 mg three times a day during six consecutive days, from the 14th to the 19th day of the estrous cycle. Plasma concentrations of progesterone (P4), androstenedione (A4), testosterone (T), estrone (E1) and estradiol-17 beta (E2) were determined by radioimmunoassay method. Infusions of Escherichia coli endotoxin resulted in a significant (p < 0.001) decrease in the production of P4, A4, T, E1 and E2 in the luteal phase as compared with the levels found in the control animals. Plasma level of P4, A4 and T was decreased by 84.6%, 86.0% and 73.0%, respectively. Plasma concentrations of E1 and E2 in some cases exceeded 5 pg/ml, nevertheless in the majority of the samples they were under sensitivity of the method. Escherichia coli endotoxin infusions resulted in a considerable decrease in the size of the ovaries, and morphological changes characteristic for acute and chronic inflammation were observed. PMID- 11944582 TI - Serological response of cattle to Chlamydophila abortus in Slovakia in 1996-2000. AB - In the Slovak Republic, in 1966-2000, 37,275 blood sera of cattle were investigated for the presence of antibodies against Chlamydophila abortus using the method of complement fixation. The antibody occurrence had following tendency: in 1996--3.72%; 1997--10.02%; 1998--9.15%; 1999--15.99%; 2000--9.51% of the tested sera contained the antibodies. In most cases, antibodies in low titres, 1:32-1:64, were detected. Positive serological reactions at such serum dilutions are not indicative of the clinical disease of cattle; they reflect an immune response of the host organism following contact with the Chlamydophila abortus antigen. The chlamydial antibody titres of 1:256, which were confirmed in 1998-1999, indicate the chlamydial infections. PMID- 11944583 TI - The connections of lymphatic vessels draining female reproductive organs with lymphnodes in the pig. AB - The connections of lymphatic vessels leaving the reproductive organs (ovary, oviduct, uterine horn, corpus and cervix, and vagina) with lymphnodes (normal and hemal) located in the uterine broad ligament and paraaortal regions were examined in the cyclic and immature pigs. To visualize the lymph pathways, the lymphatic lumen was filling with varicoloured microfil and/or ink-gelatin mass introduced all the way down to appearance in several nodes. This study revealed that lymphatics emanating from the uterine horn, corpus and the anterior part of the cervix demarcated large a lymphatic area in the porcine broad ligament. From the caudal portion of the cervix and vagina 2-3 lymphatics were found to emerge running near walls of these organs and connecting with large complex nodes, and 2 3 lymphatics reaching the plica urogenitalis. Generally, the lymphatic vessels leaving the porcine reproductive organs were connected by collector lymphangions with 3-5 normal-, 0-3 hemal- and one large composed nodes all found between the distal vascular subovarian plexus and the uterine artery as well as with 9-27 normal and 3-9 hemal nodes in the paraaortal region. PMID- 11944584 TI - CCK1 central receptor antagonist prevented the intestinal stress symptoms in sheep. AB - The aim of this study was to determine the influence of mechanically induced duodenal distension (DD) and lorglumide (CCK1 receptor antagonist) premedication on electrical activity of various parts of gastrointestinal (GI) tract and the blood plasma cortisol level in sheep. The influence of lorglumide on the unfavourable effects of duodenal distension (performed with a balloon filled with water--40 and 80 ml; DD40 and DD80) was investigated in this study. These effects in sheep were as follows: the atony of forestomachs and abomasum and the transitory stimulation of myoelectrical activity of small intestine and distal parts of large intestine. The animals, under general anaesthesia, had electrodes inserted into the muscular layers of the organ, the duodenal fistula and (in another group of animals) also the ruminal fistula. Five minute duodenal distension (DD40 and DD80) caused an immediate and complete inhibition of the frequency of spike bursts as well as reticulo-ruminal and abomasal contractions, but also a transitory significant increase of spike bursts of the intestinal wall. The duodenal distension (DD40 and DD80) caused a significant increase of plasma cortisol concentration. Lorglumide did not significantly change the motility of gastrointestinal tract and cortisol concentration, but 10 min after the intracerebroventricular (i.c.v.) infusion in the doses of 1 and 2 mg in toto (i.e. 25 and 50 micrograms/kg B.W.) it decreased the cortisol concentration by 59.7%, as compared with the control values. Lorglumide administered in the above mentioned doses 10 min before the DD40 prevented all signs of intestinal stress and decreased the release of cortisol, but only for 10 min since the beginning of the duodenal distension. It is concluded, that lorglumide--an antagonist of the central CCK1 receptors can be an effective antistressoric agent in the stomach atony caused by the duodenal distension (mechanical-algetic-emotional stress) in sheep. PMID- 11944585 TI - Diets containing leguminous seeds influence chromium content in the rat femur bone. AB - Leguminous seeds contain a lot of antinutritional factors (ANFs) such as protease inhibitors, lectins and condensed tannins which can affect bioavailability of nutrients. Detrimental effect of protease inhibitors can be decreased by sulphur amino acids addition. Moreover, most of the leguminous ANFs are thermolabile. Hence, legumes tested in our study were extruded and/or diets were supplemented with methionine and cystine (0.15% + 0.15%). The present experiment was performed for 28 days on 90 Wistar rats divided into nine feeding groups. Semipurified diets (10% casein) were supplemented (10%), except the control one, with soybean (S) or faba bean (F), raw (R) or extruded (E), with addition of sulphur amino acids (SAA) or without them. The Cr concentration was determined in the femur bone. Total feed intake during whole trial widely varied among the groups and was the lowest in SR (289 g) and FR (294 g) groups. There was no correlation between feed intake and Cr content in the femur bone. Generally, 10% soybean in the diet decreased Cr concentration in the bones, and faba bean increased it. The SAA addition to diets generally increased femur Cr content. However, there was a significant interaction between SAA supplementation and extrusion of faba bean, hence, the effect of their combination was unclear. PMID- 11944586 TI - The activity of chosen bacteriophages on Yersinia enterocolitica strains. AB - The aim of the present study was to evaluate the lytic activity of three bacteriophages on Yersinia enterocolitica strains isolated from humans and pigs. The Y. enterocolitica strains tested belonged to 0:3, 0:9 and 0:2 serogroups. The ZD5 phage was obtained from a water sample, but remaining phages were obtained from the lysogenic Y. frederiksenii 7291 and Y. enterocolitica 8684 strains. All the Y. enterocolitica strains tested which belonged to 0:9 serogroup did not show any susceptibility to the bacteriophages used. The bacteriophages tested showed different lytic activity on the Y. enterocolitica 0:3 strains investigated. The phage susceptibility of Y. enterocolitica 0:3 strains revealed 9 different phage patterns. ZD5 phage showed the highest lytic activity, because it produced confluent lysis of the most Y. enterocolitica 0:3 strains tested. The Y. enterocolitica 0:2 strains isolated from pigs showed the similar phage susceptibility. The Y. kristensenii and Y. pseudotuberculosis strains tested were not sensitive to the bacteriophages used. PMID- 11944587 TI - Effect of the gonadotrophins treatment on morphological alterations in ovary and peripheral plasma concentrations of steroid hormones in gilts. AB - The present study was designed to examine the influence of gonadotrophins treatment on the ovarian morphology changes and plasma concentrations of steroid hormones in peripheral blood. The experiment was performed on sexually pubertal gilts (Large White x Landrace) of similar age (7-8 months) and body mass (100-110 kg) with two controlled subsequent estrous cycles. The animals were randomly divided into four groups: two control consisting of pigs with the luteal phase (n = 9, the 10th day of the estrous cycle) and the follicular phase (n = 6, the 20th day of the estrous cycle) and two experimental ones consisting of animals with both mentioned periods (n = 7 and n = 9) treated with gonadotrophins (PMSG and hCG). The gilts in the luteal phase were injected (s.c.) with gonadotrophins at a daily dose of PMSG 400 and hCG 200 IU from the 16th to the 27th day (the 6th day of the next estrous cycle). The gilts in the follicular phase, were injected with the same dose of gonadotrophins but from the 8th to the 19th day of the estrous cycle. Plasma concentrations of P4, A4, T, E1, E2 and metabolite of PGF2 alpha PGFM were determined by radioimmunoassay (RIA) method. Injections of PMSG and hCG in both experimental groups produced several times enlarged: weight, size and volume of ovaries and alterations in a number of structural elements as compared with those found in the control animals. The morphological elements presented in ovaries: corpora haemorrhagica, corpora lutea, regular and atretic follicles and first of all cysts by distinctly differentiation thickness of the walls are characteristic for cystic ovarian degeneration. Plasma concentrations all determined hormones after gonadotrophins treatment in experimental groups were increased except E1 (insignificant decrease) in luteal phase as compared with those found in the control groups. Statistically significant increase (p < 0.001) in plasma concentrations of P4, A4, and T in both experimental groups and E2 (p < 0.001) in luteal phase were noted. In peripheral plasma concentrations increase of E1 and E2 in follicular phase of the estrous cycle were insignificant. PMID- 11944588 TI - Births, marriages, divorces, and deaths: provisional data for August 2001. PMID- 11944589 TI - Etizolam, an anti-anxiety agent, attenuates recurrence of chronic subdural hematoma--evaluation by computed tomography. AB - Etizolam, an anti-anxiety agent which is an antagonist of platelet-activating factor receptors, was administered to patients with chronic subdural hematoma (CSH) after hematoma removal to assess the effectiveness for preventing recurrence compared with control patients not given the drug after surgery. The remaining volumes of subdural hematomas on brain computed tomography were measured approximately 1 month after removal. Volume in the etizolam group (15 patients) was significantly smaller than in the control group (24 patients). Hematoma recurrence was not detected in the etizolam group 3 months after surgery, but occurred in the control group. The difference was significant. Etizolam administration may be useful for the prevention of recurrence of CSH. PMID- 11944591 TI - Dissecting aneurysm of the middle cerebral artery manifesting as subarachnoid hemorrhage and hemorrhagic infarctions--case report. AB - A 46-year-old woman presented with a dissecting aneurysm of the right middle cerebral artery manifesting as subarachnoid hemorrhage followed by hemorrhagic infarctions. The aneurysm was clipped and wrapped. However, serial angiography showed progression of the lesion, which was probably responsible for the clinical course of this patient. Intracranial dissecting aneurysms are less common, and the natural history of these lesions is unclear. Conservative management might be preferable in this patient. PMID- 11944590 TI - Bypass surgery for occluded internal carotid artery revascularized by vasa vasorum--case report. AB - A 47-year-old woman presented with two transient ischemic attacks due to occlusion of the left internal carotid artery. The affected artery was revascularized by vasa vasorum. Angiography showed occlusion of the left common carotid artery including the origin of the internal carotid artery. The distal internal carotid artery was revascularized by vasa vasorum at the level of the second cervical vertebral body. Left subclavian artery-internal carotid artery bypass surgery using a saphenous vein graft was performed successfully, during which the narrowed but patent lumen of the internal carotid artery was confirmed. Follow-up angiography showed enlargement of the left internal carotid artery distal to the patent bypass. Reconstructive bypass surgery is a possible treatment for patients with occluded internal carotid artery revascularized by vasa vasorum. Angiographic detection of the lumen of the internal carotid artery is important for surgical consideration. PMID- 11944592 TI - Congenital multiple cavernous angiomas associated with thrombosed arteriovenous malformation of the brain--case report. AB - A 16-year-old girl presented with multiple cerebral cavernous angiomas with calcifications due to repeated hemorrhages and a thrombosed cerebral arteriovenous malformation (AVM). Her 18-year-old elder sister also had multiple cerebral cavernous angiomas associated with calcifications, which suggested presence of repeated previous hemorrhages. Surgical removal via a right occipital craniotomy resulted in a good outcome. The histological diagnosis was thrombosed AVM. Evaluation of congenital vascular anomaly needs to take into consideration the combination of other congenital vascular anomalies and their familial occurrence. PMID- 11944593 TI - Primary rhabdomyosarcoma associated with tumoral hemorrhage--case report. AB - A 42-year-old female presented with right hemiparesis. Computed tomography (CT) demonstrated a slightly heterogeneous high density mass over the left frontal convexity with peritumoral edema, with homogeneous enhancement by contrast material. Magnetic resonance imaging showed the left frontal convexity tumor as a heterogeneous low intensity area on the T1-weighted image and a heterogeneous high intensity area on the T2-weighted image, with homogeneous enhancement and dural tail sign after intravenous administration of gadolinium diethylenetriaminepenta-acetic acid. After admission, the patient suddenly lost consciousness. CT demonstrated aggressive increase in the bulk of the tumor and large peritumoral hemorrhage. The tumor was totally removed. The histological diagnosis was rhabdomyosarcoma. Combined immunohistochemistry and electron microscopy techniques should be used to differentiate rhabdomyosarcoma from malignant meningioma. PMID- 11944594 TI - Lumbosacral metastatic extradural Merkel cell carcinoma causing nerve root compression--case report. AB - A 63-year-old man presented with a rare metastatic Merkel cell carcinoma (MCC) involving the lumbosacral spine and causing nerve root compression. Magnetic resonance (MR) imaging revealed an extradural soft tissue mass at the L5-S1 levels. The tumor was subtotally removed and chemotherapy was administered, but he died of multiple metastases from the primary epigastric tumor. Lumbosacral metastatic epidural tumor can manifest as lumbar disc disease symptoms, but MR imaging can non-invasively and rapidly reveal the presence of spinal epidural tumor and any extension to the spinal canal. Extradural MCC metastasis in the lumbosacral area should be considered in the differential diagnosis of radicular symptoms caused by disc herniation. PMID- 11944595 TI - Neuroendoscopic treatment of idiopathic occlusion of the foramen of Monro in adults--two case reports. AB - Two adults presented with hydrocephalus due to idiopathic obstruction of the bilateral foramina of Monro, manifesting as clinical signs of chronically elevated intracranial pressure. No inflammation was present. The primary surgical treatment was neuroendoscopic reconstruction of the right foramen of Monro. A 37 year-old man had a spontaneous perforation of the septum pellucidum. The patient required a ventriculoperitoneal shunt, although postoperative ventriculography proved free passage of cerebrospinal fluid from the lateral ventricle into the third ventricle. A 62-year-old man underwent additional septostomy and third ventriculostomy, and the neuroendoscopic intervention relieved the presenting symptoms without additional treatment. The biopsy specimens showed no evidence of malignancy in either case. Neuroendoscopic intervention is an alternative treatment in the management of hydrocephalus due to idiopathic obstruction of the foramen of Monor. The procedure is less invasive than open microsurgical reconstruction and can even avoid ventriculoperitoneal or ventriculoatrial shunting. PMID- 11944596 TI - Detection and characterization of the evolution of cerebral abscesses with diffusion-weighted magnetic resonance imaging--two case reports. AB - A 57-year-old man and a 45-year-old woman presented with cerebral abscesses. Diffusion-weighted magnetic resonance (MR) imaging and conventional MR imaging clearly showed the different stages of the course of the brain abscesses. As the abscess matured, the signal intensity of the center gradually increased to the typical high value with a low apparent diffusion coefficient (ADC) on diffusion weighted MR imaging, and enhancement of the capsule on T1-weighted MR imaging with gadolinium. Healing of the abscess was revealed by the signal intensity of the center returning to isointense and an increase in ADC to the baseline. Surrounding edema showed an increase in ADC, followed by a return to the baseline. These changes probably reflect the pathological processes occurring in the abscesses. PMID- 11944597 TI - Presigmoid approach for cavernous angioma in the pons--technical note. AB - Surgical treatment of brainstem lesions has been encouraged after the development of magnetic resonance imaging. However, direct approaches to intra-axial lesions in the brainstem still carry a high risk of morbidity because the neuronal structures can be injured along the entry routes. We present two patients whose pontine cavernous angiomas were removed via incision of the lateral aspect of the pons with presigmoid approach. The first case, a 41-year-old woman, presented with paresis of the cranial nerves VI, VII, and VIII, and left hemiparesis progressing over 2 weeks caused by a cavernous angioma ventrally located in the lower pons. The second case, a 50-year-old woman, developed dizziness over 2 months due to a large cavernous angioma in the center of the pons. These lesions were totally removed through the presigmoid approach and no additional neurological deficits were observed. An image-guided navigation system was used for the craniotomy and removal of the lesion in the second patient. The presigmoid approach provides a safe route to intra-axial lesions in the pons. A technique for presigmoid craniotomy with one-piece bone flap under the image guided navigation is also described. PMID- 11944598 TI - Endoscopic ventriculocystostomy through the septum pellucidum via the contralateral anterior horn--technical note. AB - Paraventricular fluid cysts have recently been treated by endoscopic fenestration performed from the cysts to the ventricular system. However, correct orientation and safe navigation of the endoscope may be difficult in patients with abnormal anatomy. Endoscopic fenestration from the ventricular system to a cyst was performed through penetration of the septum pellucidum via the anterior horn of the contralateral lateral ventricle. The advantage of this approach is correct orientation and introduction of the endoscope to the periventricular lesion because the usual landmarks can be identified in the normal contralateral lateral ventricle. PMID- 11944599 TI - Veterans Affairs bar-code-scanning system reduces medication errors. PMID- 11944601 TI - Alabama hospital pharmacists climb meritorious ladder to meet career goals. PMID- 11944600 TI - Groups revise general vaccination recommendations. PMID- 11944602 TI - Insulin glargine: basal insulin of choice? PMID- 11944603 TI - Lung cancer: a review. AB - The frequency, risk factors, pathophysiology, diagnosis, and management of lung cancer are reviewed. An estimated 157,400 Americans died of lung cancer in 2001. Lung cancer is the second most frequent cancer in both men and women. The major risk factor for lung cancer is smoking, which accounts for 75-80% of lung cancer related deaths. Lung cancers can be broadly classified into two forms, small-cell carcinomas and non-small-cell carcinomas. Non-small-cell lung cancer is more common, accounting for up to 75% of lung cancers. Lung cancer is diagnosed by chest radiography, sputum cytology, bronchoscopy, needle biopsy, and other techniques. Surgery plays a major role in managing stage I and stage II non-small cell lung cancer and may be used for stage III disease. Patients with stage IIIa disease may be surgical candidates, but involvement of mediastinal lymph nodes reduces the probability of survival. Adjuvant irradiation may reduce the rate of local recurrence but does not increase survival time. Adjuvant chemotherapy may confer a small survival-time advantage if the regimen includes cisplatin. Chemotherapy combined with radiation therapy may produce a survival advantage over irradiation alone. Patients with advanced non-small-cell lung cancer should receive combination chemotherapy. Several regimens have shown a survival advantage over best supportive care. Paclitaxel, docetaxel, gemcitabine, vinorelbine, irinotecan, and topotecan have activity both as single agents and in combination. Surgery has only a limited role in the management of small-cell lung cancer. Patients with limited disease should receive a platinum-based chemotherapy regimen plus radiation therapy. Combination chemotherapy should be offered to patients with extensive disease. The most active regimens contain cisplatin or carboplatin. Paclitaxel, docetaxel, gemcitabine, vinorelbine, irinotecan, and topotecan combinations have shown some promise. Lung cancer, although highly preventable, is usually diagnosed at an incurable stage. Chemotherapy is playing an increasingly important role alongside surgery and radiation therapy in the management of this disease. PMID- 11944604 TI - Insulin glargine: a new long-acting insulin product. AB - The pharmacodynamics, pharmacokinetics, clinical efficacy, adverse effects, and dosage and administration of insulin glargine are reviewed. Current treatment regimens for patients with type 1 diabetes mellitus and some with type 2 are designed to provide a basal insulin level with intermittent preprandial insulin coverage. Insulin glargine precipitates after subcutaneous injection, slowing absorption. Insulin glargine is used as a basal insulin and exhibits a flat pharmacokinetic profile, with a duration of action of at least 24 hours. Hypoglycemia is the most commonly reported adverse effect, especially within the first four weeks after a switch to insulin glargine. Insulin glargine should not be mixed with any other insulin product and should be administered with a syringe that has not been used for other insulin products or other medications. Insulin glargine is administered once daily at bedtime. Patients previously receiving twice-daily isophane insulin (NPH) should receive an insulin glargine dosage 20% less than the total daily dose of NPH insulin. Clinical trials did not consistently show improvements in hemoglobin A1c levels when patients with type 1 diabetes mellitus were switched from NPH insulin once or twice daily to insulin glargine. Insulin glargine should be considered for patients who continue to have elevated morning blood glucose levels and problems with nocturnal hypoglycemia despite receiving NPH insulin at bedtime. In patients with type 2 diabetes mellitus, insulin glargine significantly improved glycemic control compared with once-daily NPH insulin, but not when it was compared with combined treatment with once- or twice-daily NPH insulin. Clinical trials assessing progression of retinopathy and nephropathy and comparing insulin glargine therapy with continuous subcutaneous insulin infusion therapy are needed to more clearly determine insulin glargine's role. Insulin glargine is a new long-acting formulation that can provide prolonged basal glucose control in patients with diabetes mellitus. PMID- 11944605 TI - Efficacy of a single 8-mg i.v. dose of ondansetron hydrochloride for preventing chemotherapy-induced emesis. PMID- 11944606 TI - ASHP policy recommendations--invitation to comment. PMID- 11944607 TI - 2001 ASHP Leadership Conference on Pharmacy Practice Management Executive Summary. From management to leadership: the building blocks of professionalism. PMID- 11944608 TI - Pharmacists' role in treating hypertension. PMID- 11944609 TI - Bar coding a must for patient safety. PMID- 11944610 TI - Colony-stimulating factors: beyond the effects on hematopoiesis. AB - The effects of sargramostim and filgrastim on hematopoietic cells are described. Filgrastim is a lineage-specific colony-stimulating factor (CSF), mainly affecting neutrophils. In addition to enhancing neutrophil recovery, filgrastim may also enhance neutrophil functional activity. Filgrastim does not have any meaningful effect on monocytes or macrophages; however, recent data indicate that filgrastim has a stimulatory effect on Th2 lymphocyte-inducing dendritic cells. These dendritic cells facilitate humoral immune responses, but they also produce inhibitory cytokines that diminish cell-mediated immunity. Sargramostim is a multilineage CSF, affecting neutrophils, monocytes, macrophages, and dendritic cells. Sargramostim has a greater impact on Th1 lymphocyte-inducing dendritic cells, which facilitate cell-mediated immune responses, including antitumor activity. The broader activity of sargramostim on both types of antigen presenting cells (macrophages and dendritic cells) may account for the reports of benefit beyond enhanced neutrophil recovery that have been seen in clinical trials of patients with leukemia and patients undergoing stem-cell transplantation. Given the disparate activity of these two CSFs on the immune system and the types of immune responses generated, it is prudent for clinicians to consider these effects when choosing an agent for enhancing neutrophil recovery in various clinical settings. PMID- 11944611 TI - Formulary management of colony-stimulating factors. AB - The outcome of a formulary interchange from filgrastim to sargramostim for the amelioration of neutropenia for outpatients receiving myelosuppressive chemotherapy was evaluated. The pharmacy department at the James Graham Brown Cancer Center of the University of Louisville Hospital implemented a therapeutic interchange program by following the Joint Commission on Accreditation of Healthcare Organizations performance methodology, incorporating four key elements: plan, do, check, and act. After the pharmacy and therapeutics committee agreed that filgrastim and sargramostim are therapeutically equivalent, the pharmacy initiated the interchange, with a commitment to collect outcomes data to analyze the impact of the program on patient outcomes. Inclusion criteria included patient age of > or = 18 years, the presence of solid tumors or lymphoma, and current treatment with traditional chemotherapy. Patient demographics and cycle-specific data were collected for 31 patients receiving sargramostim and 20 patients receiving filgrastim from August 2000 to July 2001. Absolute neutrophil counts (ANCs) were measured before initiating and after discontinuing colony-stimulating factors. The majority (70%) of all growth factor use was initiated within one to four days of the last chemotherapy dose. No appreciable difference was found between agents for median ANC at any measured time point. The majority of patients exceeded the target ANC of 1500 cells/mm3 at the time of growth factor discontinuation. There were no significant differences in the number of patients that had adverse effects or in the number of cycles resulting in an adverse event between groups. Sargramostim demonstrated a 21% cost savings over filgrastim ($1036 versus $1318, respectively). The formulary switch from filgrastim to sargramostim resulted in a significant cost savings for the institution without increasing incidence of adverse effects and negative outcomes associated with growth factor use. PMID- 11944612 TI - Ten years of colony-stimulating factors: what have we learned? Introduction. PMID- 11944613 TI - Clinical applications of colony-stimulating factors: a historical perspective. AB - The development of the therapeutic roles of filgrastim and sargramostim over the past decade is reviewed. Filgrastim, a recombinant granulocyte colony-stimulating factor (G-CSF), and sargramostim, a recombinant granulocyte-macrophage colony stimulating factor (GM-CSF), have been available in the United States for over a decade to treat various types of neutropenia. In addition, data are available to support their use in a variety of clinical settings. Because of the high cost of these agents, clinical guidelines for their appropriate use have been developed. With respect to the comparability of filgrastim and sargramostim, the American Society of Clinical Oncology recommends that additional data be generated since current data are insufficient to adequately address this question in each clinical setting. A limited number of randomized, comparative trials have directly compared filgrastim with sargramostim. Outcomes data from these trials are reviewed. Further, there is evidence to suggest that, at least for GM-CSF, tolerability is dependent on the degree of protein glycosylation. A nonglycosylated protein, molgramostim, available in Europe appears to be associated with greater toxicity in clinical trials, although randomized comparative trials are lacking. The therapeutic equivalence of CSFs requires further study. While data show that the efficacy and tolerability of CSFs are similar in certain settings, there has been no definitive, randomized, large scale clinical trial conducted to address this issue. Pharmacists should continue to evaluate clinical data to determine not only which CSF to use but also when a CSF should be used. PMID- 11944614 TI - Rethinking nicotine. PMID- 11944615 TI - Kenneth Arrow and the changing economics of health care. PMID- 11944616 TI - Kenneth Arrow and the changing economics of health care. Forward. PMID- 11944617 TI - A poor prognosis for "the nursing disease". PMID- 11944619 TI - Bibliography. Current world literature. Prenatal diagnosis. PMID- 11944618 TI - Bibliography. Current world literature. Maternal-fetal medicine. PMID- 11944620 TI - Body composition and its measurement in vivo. 1967. PMID- 11944621 TI - Roundtable discussion: hypertension, renal disease, and diabetes. AB - Following a symposium on Hypertension in Atlanta, Georgia on April 25, 2001, which was sponsored by the National Heart, Lung, and Blood Institute, a panel was convened to discuss the problems of renal disease and hypertension. PMID- 11944622 TI - Spontaneous cholesteatoma of the external auditory canal. AB - Spontaneous cholesteatoma of the external auditory canal is a rare disease which may also be under diagnosed. Few cases have been reported even in major otology centers. Possible theories as to the aetiology are based on observation of these few cases. The correct diagnosis allows the proper treatment avoiding the progression of the cholesteatoma and further hearing loss or other complications. The authors review five cases and discuss the possibility that it is the anatomic structure of the bony external auditory canal bone that might contribute to the etiopathogenesis of these cholesteatomas. PMID- 11944623 TI - [Ciliary neurotrophic factor and nerve regeneration]. PMID- 11944624 TI - ["Research training in neurosurgery residency programs. Analysis of the situation in other developed countries and proposals to complement the current program in Spain"]. PMID- 11944625 TI - [Grant for a resident to study abroad]. PMID- 11944627 TI - Voluntary health workers' knowledge, attitude and practices regarding record keeping in Akinyele LGA of Oyo State, Nigeria. AB - Lack of basic health data for monitoring and evaluation of health services continue to affect the planning, implementation and evaluation of health services in Nigeria. This has contributed to the poor health status and inefficient health services in the country. In the primary health care management information system (PHCMIS), Voluntary Village Health Workers (VHWs) and Traditional Birth Attendants (TBAs) are responsible for collecting information at the community/village level. Trained research assistants administered a pretested semi structured questionnaire and filled an observation checklist to all the voluntary health workers in Akinyele LGA of Oyo State in a cross-sectional survey conducted to assess their record keeping practices as well as their knowledge of and attitude towards it. Results showed that almost half of the respondents had no formal education and a similar proportion had been VHWs for between over 10 years. Over eighty per cent knew the uses of record keeping for monitoring and evaluation purposes. Their attitude towards it was positive and almost all felt it was easy to keep records. Ninety six percent keep records of their health activities and most forward them. It was observed that only 11 (10.8%) respondents had the VHW/TBA record of work produced and recommended by the Federal Ministry of Health because they were reportedly not supplied to them. The factors that were associated with record keeping practices were positive attitude towards record keeping, duration of work as a VHW/TBA, prior training on record keeping and receiving feedback on records kept. Recommendations made included periodic training and retraining of the VHWs on record keeping, ensuring consistent supply of record forms and providing regular feedback on records kept to the VHWs. PMID- 11944628 TI - What is ISMICS and why does it exist? PMID- 11944629 TI - The use of stentless valves for root replacement during repair of ascending aortic aneurysms with aortic valve regurgitation. AB - BACKGROUND: Early and mid-term results of stentless valves for the treatment of ascending aortic aneurysm (AAA) were evaluated in a retrospective manner. MATERIAL AND METHODS: During a four-year period, 26 patients with ascending aortic aneurysms and aortic valve insufficiency underwent a total root replacement procedure using a stentless "Freestyle" valve (Medtronic Inc., Minneapolis MN). Mean age was 71 +/- 4 years (range 66 to 79 years). Eight patients were in NYHA Class 2, 13 in Class 3, and 5 in Class 4. Cardiopulmonary bypass (CPB) was begun with femoral artery-right atrium (two-stage) cannulation in all cases but four, in which the right axillary artery was used. Myocardial protection was established by retrograde, cold-blood cardioplegia and direct antegrade blood cardioplegia from the right coronary ostium. The left ventricle outflow tract was constructed by using 2-0 ticron sutures and incorporating a pericardial strip in between. Coronary buttons were sewn to the xenograft with 6 0 polypropylene sutures. Meanwhile, the patient was cooled down to 18 degrees nasopharyngeal temperature and the distal anastomosis with the proximal arch was performed with a Dacron graft under total circulatory arrest (TCA), using 4-0 polypropylene sutures. During rewarming, the connection between the Freestyle valve and the Dacron graft was performed. RESULTS: Ischemic time was 91 +/- 11 minutes and TCA time was 9 +/- 4 minutes. Operative mortality was zero, and there was one 30-day mortality (3.8%). At discharge, all 25 patients had a functional valve with low transvalvular gradients. Patients were followed for a mean period of 15 months, with one patient being lost to follow-up and one patient dying of non-cardiac causes. Follow-up was 97% complete, and echocardiographic control during the follow-up period revealed competent valves with gradients comparable to those at discharge. Two patients were screened with electron beam tomography (EBT) three years after the operation and there was no sign of wall or leaflet calcification. At the end of the 15 months (mean) follow-up, the functional capacity of the patients had improved significantly (p <0.05). CONCLUSIONS: Our early results suggest that use of the Freestyle valve in conjunction with a Dacron tube graft can be a good alternative for patients over 65 years of age who present with ascending aortic aneurysm with aortic valve insufficiency. PMID- 11944630 TI - Diabetes mellitus, obesity, and hepatic steatosis. AB - Nonalcoholic fatty liver disease is emerging as the most common liver disease in North America. The histological spectrum of nonalcoholic fatty liver disease ranges from fatty liver alone to steatohepatitis and to the most serious form- nonalcoholic steatohepatitis (NASH). An increasing body of evidence suggests that NASH is associated with the development of progressive fibrosis and eventually cirrhosis in approximately 20% of cases. These data emphasize the need to develop effective therapy for the treatment of NASH. Cases occur most commonly in obese middle age women with diabetes. However, NASH may also occur in children and normal weight men with normal glucose and lipid metabolism. The pathophysiology involves 2 steps. The first is insulin resistance, which causes steatosis. The second is oxidative stress, which produces lipid peroxidation and activates inflammatory cytokines resulting in NASH. Liver biopsy provides prognostic information and identifies NASH patients who may benefit from therapy. Treatment consists of managing the comorbidities: obesity, diabetes, and hyperlipidemia. Nascent clinical trials suggest that a number of therapies may be beneficial. These include anti-oxidants such as vitamin E and betaine, bile acid therapy with ursodeoxycholic acid, and improved insulin sensitivity with metformin. Another potential therapeutic strategy is the reduction of inflammatory cytokines. PMID- 11944631 TI - Fat and the liver--a molecular overview. AB - Nonalcoholic fatty liver disease (NAFLD) is a chronic liver disease that occurs in nondrinkers but which cannot be distinguished from alcohol-induced liver disease histologically. There are no diagnostic blood tests for NAFLD but the disease is associated with several insulin-resistant states, including obesity, type 2 diabetes mellitus and dyslipidemia. Most of the liver-related morbidity and mortality that accompany NAFLD occur in patients who develop cirrhosis. The latter is most likely to occur in individuals who have progressed from simple steatosis (fatty liver) to steatohepatitis, a chronic inflammatory liver lesion. The mechanisms that promote the transition from steatosis to nonalcoholic steatohepatitis appear to involve multiple cellular adaptations to the oxidative stress that occurs when fatty acid metabolism is deranged during insulin resistance. A better understanding of these mechanisms is desired to target treatments to prevent and/or reverse nonalcoholic steatohepatitis, thereby aborting the evolution of cirrhosis. PMID- 11944632 TI - Alcoholic steatosis and steatohepatitis. AB - Hepatic steatosis and steatohepatitis are encountered with great frequency in people who consume large amounts of ethanol (more than 6 drinks per day). Ethanol causes steatosis by altering several steps in the hepatic processing of fatty acids, including their uptake from plasma, their use as fuel substrates, and their export as triglyceride. When clinically mild, alcoholic steatosis and steatohepatitis can be difficult to distinguish from nonalcoholic fatty liver disease. This is particularly true among individuals at high risk of accelerated alcoholic liver injury, such as women, the obese, and those with hepatitis C. In the outpatient setting, history and aspartate aminotransferase:alanine aminotransferase ratio offer the best clues to diagnosis. Liver biopsy cannot determine the cause of steatohepatitis, but can show the extent of disease. The etiology of disease is important to prognosis, as alcoholic fatty liver carries a much higher risk of progression and mortality than nonalcoholic fatty liver disease. Patients with moderate to severe alcoholic steatohepatitis are typically hospitalized. Derangements in white blood cell count, prothrombin time, and bilirubin identify those with the highest early mortality. Survival in this severely ill subgroup is improved with the short-term use of corticosteroids; patients who have contraindications to steroids may benefit from other forms of therapy, either pharmacologic, nutritional, or both. PMID- 11944633 TI - Hepatitis C and steatosis. AB - Hepatic steatosis is common in patients with chronic hepatitis C virus (HCV) infection. Epidemiologic studies have shown HCV-associated steatosis to correlate with both patient factors, such as obesity and viral factors, such as HCV genotype 3a. Furthermore, the degree of steatosis has been linked to the extent of hepatic fibrosis in several studies, implying that steatosis may be contributing to disease progression in chronic HCV infection. Whether the pathogenesis of HCV-associated steatosis is linked to oxidative damage non specifically, to HCV viral properties, or to other factors remains unknown. This steatosis may play an important role in the response to HCV therapy, in disease progression after liver transplantation for HCV, or in HIV-HCV coinfection. PMID- 11944634 TI - The fatty liver in AIDS. AB - Steatosis or fatty liver in individuals with human immunodeficiency virus (HIV) may result from HIV itself, the use of nucleoside analogues, concurrent infection with hepatitis B or C, alcohol use, diabetes mellitus, obesity, or combinations of these factors. Nucleoside analogues have been the focus of increasing concern, because several fatal cases of severe macrosteatosis, lactic acidosis, and hepatomegaly have been linked to the use of nucleoside analogues. Other classes of antiretroviral drugs, as well as opportunistic infections, can also cause hepatic injury without steatosis. The additive effect of these different risk factors, especially in the presence of underlying hepatic steatosis, likely contributes to the increased prevalence of hepatic abnormalities among HIV infected individuals. The conditions under which some patients rapidly progress to hepatic failure and/or cirrhosis need to be defined. This is a US government work. There are no restrictions on its use. PMID- 11944636 TI - A long-suffering patient with new abdominal pain. PMID- 11944635 TI - Mitochondrial fatty acid oxidation and acute fatty liver of pregnancy. AB - Accumulation of micro- and macrovesicular fat in hepatocytes is the hallmark of several liver diseases of pregnancy, including acute fatty liver of pregnancy (AFLP) and severe pre-eclampsia. While the exact origin of AFLP is not known, this pathologic finding and its similarity to the hepatopathology found in infants and children with inborn errors of mitochondrial fatty acid-oxidation suggests a common pathogenesis. Over the last few years, the incidence of AFLP and other severe complications of pregnancy have been found to be increased in women who carry a fetus with a defect in fatty acid oxidation, and who are themselves carriers of a genetic mutation that partially compromises their own intramitochondrial fatty acid oxidation pathway. This article reviews evidence that links AFLP with both genetic and acquired factors that compromise intramitochondrial fatty acid oxidation. Based on this review, recommendations are offered for obstetric, perinatal, and neonatal management of affected mothers and infants. PMID- 11944637 TI - Final rules relating to use of electronic communication and recordkeeping technologies by employee pension and welfare benefit plans. Notice of final rulemaking. AB - This document contains final rules under Title I of the Employee Retirement Income Security Act of 1974, as amended (ERISA), concerning the disclosure of certain employee benefit plan information through electronic media, and the maintenance and retention of employee benefit plan records in electronic form. The rules establish a safe harbor pursuant to which all pension and welfare benefit plans covered by Title I of ERISA may use electronic media to satisfy disclosure obligations under Title I of ERISA. The rules also provide standards concerning the use of electronic media in the maintenance and retention of records required by sections 107 and 209 of ERISA. The rules affect employee pension and welfare benefit plans, including group health plans, plan sponsors, administrators and fiduciaries, and plan participants and beneficiaries. PMID- 11944638 TI - FDA's foods program. PMID- 11944639 TI - The role of the state as a drug purchaser. PMID- 11944640 TI - The vulnerability of HIPAA regulations to First and Fourth Amendment attack: an addendum to "Evolving Constitutional Privacy Doctrines Affecting Healthcare Enterprises". PMID- 11944641 TI - Federal regulation of single-use medical devices: a revised FDA policy. PMID- 11944642 TI - The evolution of substantial equivalence in FDA's premarket review of medical devices. PMID- 11944643 TI - PMA primacy: synthesizing the 35 U.S.C. Section 156 patent term extension, 35 U.S.C. Section 271(e)(1) patent infringement exemption as currently applied to medical devices, and medical device preemption jurisprudence to yield a cohesive solution regarding scope of coverage. PMID- 11944644 TI - The National Childhood Vaccine Injury Act turns fifteen. PMID- 11944645 TI - Compensation versus colonization: a common heritage approach to the use of indigenous medicine in developing Western pharmaceuticals. PMID- 11944646 TI - The memorial to the fallen of the Medical Corps. PMID- 11944647 TI - [Establishment of public health in Japan and Britain: the roles played by Sensai Nagayo and Edwin Chadwick]. AB - Both Japan and Britain established public health administration as a result of the similar events: cholera epidemics. Britain was the pioneer in enacting public health laws in the world. The British law was based upon the "sanitary report" compiled by E. Chadwick as part of a government committee investigation. Though the process was not always a smooth one, Britain learned from its mistakes and steadily developed a public health system following the establishment of the public health law. Meanwhile, in Japan where the general understanding is that the health care system before World War II was German-oriented, Sensai Nagayo, the leading figure in health administration, looked toward the British model and consistently attached importance to a public health administration led by local authorities. However, his vision was not realized. In this paper, I have examined what caused the differences between these two cases through focusing on the role the two key persons played. PMID- 11944648 TI - [A study of the organizing process of the modern midwifery system in Yamagata Prefecture focusing on the enactment of "Midwifery Regulation" in the 32th year of Meiji]. AB - The Japanese modern midwifery system was provided at the start by the "Medical regulation" in the 7th year of Meiji and was organized to national unity by the Imperial Ordinance "Midwifery Regulation" in the 32th year of Meiji (1899). During these twenty-five years, Hokkaido and each of the prefectures enacted their own "Midwifery Regulation" to conform with the "Midwifery Regulation," and they organized the original midwifery system. This study focuses on the Yamagata Prefecture and midwifery system in the Tohoku district, and considers the organizing process of the system. Because the Yamagata district continued the Edo Period custom of infanticide, the Yamagata prefectual administration maintained strict control of that custom. At the same time it repeatedly enacted and revised the "Mid-wifery Regulation" to adjust it to the conditions of farm and mountain villages. Finally, it established the original "Registered Midwife System" in the 22th year of Meiji. That organizing process was classified into four stages. The present study shows that the system was not contradictory to the Imperial Ordinance "Midwifery Regulation" enacted in the 32th year of Meiji, which was amended to the unified national midwife system. PMID- 11944649 TI - [Theories of medicine in the newly discovered medical book "Hon'po Kagen Hishu" by San'ki Tashiro: basic prescriptions and modified prescriptions]. AB - While investigating the formation of the Gosei-ha school in Japan by Tashiro San'ki and Manase Dosan, we discovered "Hon'po Kagen Hishu", a heretofore unknown medical book by San'ki Tashiro. A comparison of this work with Sanki's "Wakyokushu" revealed the following points. (1) "Hon'po Kagen Hishu" is a medical book compiled by San'ki Tashiro, and is believed to have been put together after the newly-compiled "Wakyokushu" (1525). (2) The book is roughly divided into two parts: basic prescriptions and modified prescriptions. "Hon'po Kagen Hishu" reveals clearly that San'ki's medical theories form the key theories of these two parts. (3) Many of the basic prescriptions contained in "Hon'po Kagen Hishu" were existing prescriptions used by the Kyokuho-ha School which was popular in Japan at the time. The modified prescriptions conform to the bian zheng pei ji (selection of drug based on the differential diagnosis) theories of the Li-Zhu medical school. (4) Among the 15 prescriptions described in the 'Shobyou Tsuyo no Yakuho' chapter of "Hon'po Kagen Hisku" secret prescriptions from schools of thought including the Nakarai school were discovered, pointing to a connection between San'ki and the Nakarai school. PMID- 11944650 TI - [A study on the process and significance of the establishment of the nurse regulations in 1915]. AB - Believing that the origin of the complexity of the present nurse education and training courses lies in the nurse regulations enacted by the Ministry of Home Affairs in 1915, the author has studied the background and significance of the regulations through investigations and analyses of the regulations which were promulgated in prefectures prior to the 1915 nurse regulations. This study reveals that firstly, the nurse regulations made by the Ministry of Home Affairs were enacted in the intensive form of regulations locally promulgated prior to the nurse regulations, secondly that the qualification structure for nurses was the same as that for doctors, and thirdly that the two ways of nurse license acquisition; by either qualification exams or graduation from training schools, and the two level license systems; namely registered and enrolled nurses, were all decided by the nurse regulations and were the origin of the complexity of the present nurse systems, which have influenced the public health nurse midwife and nurse regulations promulgated after the second world war. PMID- 11944651 TI - [From Dokushoan Nagatomi to Genshun Koishi]. AB - In the middle part of the Edo period (around the later half of the 18th Century), several new-ideated physicians did appear. Among them, Dokushoan Nagatomi and Genshun Koishi were eager to learn western style autopsy. The former got the knowledge from imported Dutch books with the aid of interpreters at Nagasaki and the latter, upon influence of the former, eventually performed on autopsy by himself, aiming at understanding relationships between organ changes and the nature of the diseases of the patients. This article describes an outline of the story of two physicians as pioneers of anatomy and pathological anatomy in this country. PMID- 11944652 TI - Genetic testing under the ADA: a case for protection from employment discrimination. PMID- 11944653 TI - [Secret medicines]. AB - Secret medicines had two characteristics: their formula remained unknown and they were prepared by many kinds of people. Before 1728 there were no general laws about these secret medicines but only peculiar rules. From 1728 to 1778, the King edicted rigorous rules in order to limit the number of secret medicines. Between 1778 and 1789, the law became more definite and the Royal Society of Medicine gave advices. The Law of Germinal An-XI forbid secret medicines but since 1805, some compromises took place. Slowly, secret medicines were replaced by pharmaceutics and new set of laws. PMID- 11944654 TI - [Wine and alcohol in apothecaries' shops during the Middle Ages in Southern countries]. AB - Alcohol, wine and their derivatives do not play during the Middle Ages the role they played during Antiquity. The reading of ancient authors by Arab doctors was probably at the origin of this lack of interest for wine. As ordinary wine, medicinal wines were a matter of conviviality more than of therapeutics. Vinegar was more often used, maybe because of its lack of inebriating properties. Alcohol, this mysterious product, was probably more pertinent in the area of alchemy. Doctors and pharmacists of the enlightened age gave it a new importance. The influence of Islam on Middle Age medicine in Christian occident could explain this lack of interest for wine. PMID- 11944655 TI - [History of the Naval School of Anatomy and Surgery of Rochefort (1722-1964)]. AB - The Ecole d'anatomie et de chirurgie navale de Rochefort was created on February 5, 1722 by Jean Cochon-Dupuy. Anatomy, internal medicine and botany were taught together with the art of bandages. The major professor was the first medicine doctor of the port, afterwards came the first surgeon. At the end of the XVIIIth century the number of subjects and teachers increased. Many terms of probation were also organized. This school was closed in 1964. PMID- 11944656 TI - [Gods, women and pharmacy in Greek Mythology]. AB - The study of Greek Mythology fully justifies Herophilus's phrase: "Medicines are the hands of Gods" (third cent. B.C.). A number of Gods are said to be the inventors of the drugs which are useful to men. Their names are still alive in the scholarly or popular appellations of a great many medicinal herbs. However, insofar as the action of a drug (of a Pharmakon) remains mysterious, one finds it in essentially female practices as well as in medicine. The study of these ancient beliefs, which have survived in spite of the progress of twentieth century science, can develop the history of epistemology of pharmacy by stimulating interdisciplinary research. PMID- 11944657 TI - [Jean Hachette, pharmacist of Maison royale de Sante, complementary biography]. AB - Until 1814, Jean Hachette (1775-1840) practised as a military pharmacist and then as a hospital pharmacist in Paris. For him both professional and personal aspects of his life were affected by his fondness for gambling. His son, Louis (1800 1860), famous businessman and publisher, tempted to introduce him again to the authorities as military pharmacist in 1830 when he had to resign after a dispute with professeur Dumeril (1774-1860). So, Hachette ended his career at the military hospital of Longwy. These facts are attested by archives documents. PMID- 11944659 TI - [The Swiss Commission of Goiter of January 21, 1922]. AB - The Swiss Commission of Goiter met for the first time the 21st of January, 1922 and has been the first step of an historical event in terms of Public Health: the first requirement for salt iodination for the prevention of goiter. The minutes of this commission as well as the following scientific publications until 1930 indicate that the debate was still very vigorous about this recommendation but mainly about the origin of goiter and the iodine dose to be used. This dose went progressively from less than 10 mg of iodine per kilo of salt to the present WHO recommendation of 20 to 40 mg of iodine per kilo of salt. Another key point was the strategy of implementation of iodine. Two complementary approaches were proposed and implemented: iodine supplement to children at school and iodized salt available for the whole population. One point seemed clear for everyone at that time: iodine was not the source of goiter. Later scientific studies have shown that iodine deficiency was indeed the origin of goiter and associated pathologies. PMID- 11944658 TI - [Projection and representation of scientific France in the Eastern Portuguese Empire: the correspondence exchanged between the Boletim General de Medicina e Pharmacia of the medical-surgical School of Nova Goa and the Faculty of Medicine of Lille]. AB - This article approaches the projection and representation of scientific France in the Eastern Portuguese Empire. It deals, more precisely, with the correspondence exchanged between the Boletim Geral de Medicina e Pharmacia of the Medical Surgical School of Nova Goa and the Faculty of Medicine of Lille. The ancient colony of Portuguese India presented a certain tradition in the publishing of medical-pharmaceutical magazines in the period between the second half of the nineteenth periodical approached certain themes of professional nature and included some articles of scientific nature with a special emphasis to certain aspects of tropical medicine and pharmacy. In Boletim Geral de Medicina e Pharmacia, it is possible to find the influence of French scientific culture on that ancient Portuguese culture. The exchange of correspondence between the Medical-Surgical School of Nova Goa and the Faculty of Medicine of Lille after World War I shows the solidarity which existed between these two institutions. It also shows the high consideration showed by the School of Nova Goa to the Faculty of Medicine of Lille and to French science and culture in general, universally recognised by the creative capacity of innovating in the service of common well being. PMID- 11944660 TI - Manufacturing convictions: why defendants are entitled to the data underlying forensic DNA kits. PMID- 11944661 TI - Forcible medication of mentally ill criminal defendants: the case of Russell Eugene Weston, Jr. PMID- 11944662 TI - Livestock ammonia management and particulate-related health benefits. AB - Agricultural operations are the largest source of ammonia emissions in the United States and contribute to the formation of ammonium nitrate and ammonium sulfate, two prevalent forms of fine particulate matter. Researchers have found an association between fine particulate matter and a variety of adverse healths effects, including premature mortality, chronic bronchitis, hospital admissions, and asthma attacks. Management practices that reduce ammonia emissions may decrease adverse health effects, resulting in significant economic benefits. We estimated the impact of a variety of emission controls, including diet optimization, alum, and incorporation of manure into the land. The results suggest that relatively modest management policies can have a significant impact on fine particulate formation in the atmosphere. Because of the heterogeneous nature of particulate matter, a key question is the importance of particulate matter size and composition. To the extent that ammonium nitrate and ammonium sulfate contribute to adverse health effects, ammonia management may have significant health implications. Our results suggestthat a 10% reduction in livestock ammonia emissions can lead to over $4 billion annually in particulate related health benefits. PMID- 11944663 TI - Including transformation products into the risk assessment for chemicals: the case of nonylphenol ethoxylate usage in Switzerland. AB - A method for applying the risk assessment approach using ratios of predicted environmental concentrations (PECs) and predicted no-effect concentrations (PNECs) to mixtures of parent compounds and their environmental transformation products is presented. Nonylphenol ethoxylates (NPnEOs) and a selection of their most relevant transformation products are investigated as a case study illustrating the method. The PEC values of NPnEO and its transformation products are calculated with a regional multimedia fate model including the transformation kinetics of the NPnEO degradation cascade. PNEC values are derived from a selection of toxicity data on NPnEO and its transformation products. The toxicity of the emerging mixture of NPnEO and its transformation products is then estimated under the assumption of concentration addition (similar mode of action). On this basis, PEC-to-PNEC ratios for the aquatic environment and the sediment are calculated for the individual components of the mixture and the mixture itself. For this purpose, average release rates of NPnEO and its transformation products from Swiss sewage treatment plants were used. While the PEC values of the individual components do not exceed the corresponding PNEC values, the risk quotient of the mixture in water is greater than 1. In sediment, the mixture does not exceed a risk quotient of 1. A combination of sensitivity and scenario analyses is employed to identify the upper and lower bounds of the results. PMID- 11944664 TI - Semivolatile organochlorine compounds in the free troposphere of the Northeastern Atlantic. AB - Polychlorobiphenyls (PCBs), hexachlorobenzene (HCB), hexachlorocyclohexanes, and DOTs were analyzed over 1 entire year period in the air of a high altitude remote site (2367 m above sea level) located in Teide (Tenerife, Canary Islands, 28 degrees N16 degrees W) in the Eastern North Atlantic region. Twenty samples were collected providing information on the concentrations of these semivolatile organochlorine compounds (SOCs) in the free troposphere since the stable and persistenttemperature inversion in the subtropics defines a clear separation from the marine boundary layer. More than 80% of total SOCs were in the gas phase. HCB was the individual SOC in higher concentration, 51 pg m(-3), well above than the other SOCs identified, 1-11 pg m(-3). Sum PCB concentrations were 78 pg m(-3). These concentrations range among the lowest described in atmospheric samples. The collected air originated from four main sectors, high and middle latitudes in the north Atlantic, Europe, and Africa, as determined from isentropic backward air mass trajectories. No significant differences were observed for the concentrations of these compounds between air masses showing a high uniform SOC composition of the free troposphere. Only the more volatile PCB congeners, #18 and #28, exhibited significant differences between air masses from northern and southern latitudes. A seasonal temperature dependence for the less volatile PCB congeners, five or more chlorine subtituents, was also observed. Free tropospheric concentrations of all SOCs except HCB were lower than those measured near sea level for reference. PCB concentrations at the two altitudes exhibited consistent differences according to degree of chlorination. The stronger decrease of the more volatile compounds with altitude might reflect higher photodegradation. PMID- 11944665 TI - Atmospheric gas-particle partitioning of polycyclic aromatic hydrocarbons in high mountain regions of Europe. AB - Atmospheric concentrations and gas-particle partitioning of polycyclic aromatic hydrocarbons (PAH) have been determined at three remote mountain areas in Europe. Gas-phase mean concentrations of total PAH (20 individual compounds) were very similar at all sites, ranging from 1.3-2.6 ng m(-3) in the Pyrenees (Spain) to 2.7-3.7 ng m(-3) in the Alps (Austria) and Caledonian mountains (Norway). A seasonal variability was observed, with the highest levels found in winter. The seasonal differences were reflected better in the particle-associated PAH, showing the increase of PAH emissions in the colder months and a temperature dependence of the gas-particle partitioning. Significant geographical differences were also observed for particulate PAH, indicating a greater influence of regional sources than in the gas phase. Partitioning of PAH between gas and particulate phases was well-correlated with the subcooled liquid vapor pressure in all samples, but with slopes significantly steeper than the expected value of 1. These steeper slopes may reflect the occurrence of a nonexchangeable PAH fraction in the aerosols, likely associated to the soot carbon phase. Comparison of absorption to organic matter and soot carbon using the octanol-air (Koa) and soot-air (Ksa) partitioning coefficients shows that, despite uncertainties on estimated organic matter and soot carbon contents in the sampled aerosols, Koa underpredicts aerosol PAH concentrations by a factor of 0.6-2 log units. In contrast, predicted and measured high mountain aerosol PAH differ by 0.2-0.6 log units when Ksa is considered. The results point to soot carbon as the main transport medium for the long-range distribution of aerosol-associated PAH. PMID- 11944666 TI - Measurement of emissions from air pollution sources. 5. C1-C32 organic compounds from gasoline-powered motor vehicles. AB - Gas- and particle-phase organic compounds present in the tailpipe emissions from an in-use fleet of gasoline-powered automobiles and light-duty trucks were quantified using a two-stage dilution source sampling system. The vehicles were driven through the cold-start Federal Test Procedure (FTP) urban driving cycle on a transient dynamometer. Emission rates of 66 volatile hydrocarbons, 96 semi volatile and particle-phase organic compounds, 27 carbonyls, and fine particle mass and chemical composition were quantified. Six isoprenoids and two tricyclic terpanes, which are quantified using new source sampling techniques for semi volatile organic compounds, have been identified as potential tracers for gasoline-powered motor vehicle emissions. A composite of the commercially distributed California Phase II Reformulated Gasoline used in these tests was analyzed by several analytical methods to quantify the gasoline composition, including some organic compounds that are found in the atmosphere as semi volatile and particle-phase organic compounds. These results allow a direct comparison of the semi-volatile and particle-phase organic compound emissions from gasoline-powered motor vehicles to the gasoline burned by these vehicles. The distribution of n-alkanes and isoprenoids emitted from the catalyst-equipped gasoline-powered vehicles is the same as the distribution of these compounds found in the gasoline used, whereas the distribution of these compounds in the emissions from the noncatalyst vehicles is very different from the distribution in the fuel. In contrast, the distribution of the polycyclic aromatic hydrocarbons and their methylated homologues in the gasoline is significantly different from the distribution of the PAH in the tailpipe emissions from both types of vehicles. PMID- 11944667 TI - Episodic particle transport events controlling PAH and PCB cycling in Grand Traverse Bay, Lake Michigan. AB - To evaluate the influence of episodic events on particle and hydrophobic organic contaminant (HOC) cycling in the Great Lakes, we deployed sequencing sediment traps at two locations in the western arm of Grand Traverse Bay, Lake Michigan. The traps collected integrated samples of settling particles every 2 weeks from May 1997 to September 1999. The total polycyclic aromatic hydrocarbon (t-PAH) and total polychlorinated biphenyl (t-PCB) settling fluxes from the surface waters in the southern site were significantly greater than those from the northern site. In addition, there were more frequent brief increases in the mass flux to the southern site than to the northern site. These episodic events, which occurred only 20% of the time, accounted for 65% of both the mass flux and t-PAH flux. The t-PCB flux was not influenced by these episodic events, and only 18% of the t-PCB flux occurred during these events. PAHs and PCBs appear to be tracing different types of particles in the water column. Several large mass flux events characteristic of seiches were observed simultaneously in the benthic nepheloid layer (BNL) at both the northern and the southern sites. The particles settling as a result of these resuspension events had lower t-PCB and t-PAH concentrations than particles settling at other times. This suggests that the material settling into the traps on the high mass flux days is composed of a mixture of the less contaminated underlying resuspended sediment and the "regular" contaminant-rich particles settling into the BNL. PMID- 11944668 TI - Comparison of black smoke and PM2.5 levels in indoor and outdoor environments of four European cities. AB - Recent studies on separated particle-size fractions highlight the health significance of particulate matter smaller than 2.5 microm (PM2.5), but gravimetric methods do not identify specific particle sources. Diesel exhaust particles (DEP) contain elemental carbon (EC), the dominant light-absorbing substance in the atmosphere. Black smoke (BS) is a measure for light absorption of PM and, thus, an alternative way to estimating EC concentrations, which may serve as a proxy for diesel exhaust emissions. We analyzed PM2.5 and BS data collected within the EXPOLIS study (Air Pollution Exposure Distribution within Adult Urban Populations in Europe) in Athens, Basel, Helsinki, and Prague. 186 indoor/outdoor filter pairs were sampled and analyzed. PM2.5 and BS levels were lowest in Helsinki, moderate in Basel, and remarkably higher in Athens and Prague. In each city, Spearman correlation coefficients of indoor versus outdoor were higher for BS (range rspearman: 0.57-0.86) than for PM2.5 (0.05-0.69). In a BS linear regression model (all data), outdoor levels explained clearly more of indoor variation (86%) than in the corresponding PM2.5 model (59%). In conclusion, ambient BS seizes a health-relevant fraction of fine particles to which people are exposed indoors and outdoors and exposure to which can be assessed by monitoring outdoor concentrations. BS measured on PM2.5 filters can be recommended as a valid and cheap additional indicator in studies on combustion related air pollution and health. PMID- 11944669 TI - Detection of volatile organometal chloride species in model atmosphere above seawater and sediment. AB - The principal route for release of organometallic species into the atmosphere is currently considered to be the biological or chemical formation of saturated volatile compounds such as dimethylmercury, metal hydrides, etc. Model experiments conducted in our laboratory pointto the release of extremely toxic, volatile organotin, -lead, -mercury, and -arsenic species that have been detected as the chlorides in the gas phase above seawater and sediment mixtures. As the sediment was sterilized prior to the analytical study, any bioformation of volatile metal species was excluded. The most probable account of these observations is the naturally occurring formation of metal chlorides, fueled by the abundance of chloride ion (or other halides) present in seawater and many other natural environments. This raises the suggestion that the release of toxic trace elements to the atmosphere by environmental processes may be seriously underestimated. A conservative projection, based on laboratory data, suggests that the emission of butyltin into the atmosphere may reach as high as 150 ng/m2 of water/yr from a polluted marine environment, suggesting a very significant level of release of butyltin from seawater at the global level. For the sampling and determination of volatile metal species, solid-phase microextraction combined with inductively coupled plasma time-of-flight mass spectrometry has been used. This new analytical system is capable of detection of semivolatile, thermally unstable compounds such as the class encompassing metal halide species. PMID- 11944670 TI - Pharmaceuticals, hormones, and other organic wastewater contaminants in U.S. streams, 1999-2000: a national reconnaissance. AB - To provide the first nationwide reconnaissance of the occurrence of pharmaceuticals, hormones, and other organic wastewater contaminants (OWCs) in water resources, the U.S. Geological Survey used five newly developed analytical methods to measure concentrations of 95 OWCs in water samples from a network of 139 streams across 30 states during 1999 and 2000. The selection of sampling sites was biased toward streams susceptible to contamination (i.e. downstream of intense urbanization and livestock production). OWCs were prevalent during this study, being found in 80% of the streams sampled. The compounds detected represent a wide range of residential, industrial, and agricultural origins and uses with 82 of the 95 OWCs being found during this study. The most frequently detected compounds were coprostanol (fecal steroid), cholesterol (plant and animal steroid), N,N-diethyltoluamide (insect repellant), caffeine (stimulant), triclosan (antimicrobial disinfectant), tri(2-chloroethyl)phosphate (fire retardant), and 4-nonylphenol (nonionic detergent metabolite). Measured concentrations for this study were generally low and rarely exceeded drinking water guidelines, drinking-water health advisories, or aquatic-life criteria. Many compounds, however, do not have such guidelines established. The detection of multiple OWCs was common for this study, with a median of seven and as many as 38 OWCs being found in a given water sample. Little is known about the potential interactive effects (such as synergistic or antagonistic toxicity) that may occur from complex mixtures of OWCs in the environment. In addition, results of this study demonstrate the importance of obtaining data on metabolites to fully understand not only the fate and transport of OWCs in the hydrologic system but also their ultimate overall effect on human health and the environment. PMID- 11944671 TI - Macroscopic studies of the effects of selenate and selenite on cobalt sorption to gamma-Al2O3. AB - Metal ion sorption can be significantly impacted by the presence of other solutes or complexing species. In this research, macroscopic sorption studies were conducted to evaluate the effect of strongly sorbing Se(IV) and weakly sorbing Se(VI) oxyanions on cobalt(II) sorption to gamma-Al2O3. Se(IV) was found to significantly alter Co(ll) sorption as a function of Co(II) surface coverage, while Se(VI) was found to have no effect on Co(II) sorption. Under low Co(II) surface loadings (<0.1 micromol/m2), Se(IV) increased Co(II) sorption as a function of the Se(IV) coverage. At low Se(IV) surface coverages, no change in Co(II) sorption was detectable, while at high Se(IV) loadings Co(II) sorption was significantly increased. The increase in Co(ll) sorption in the bisorbate systems can be explained by either an electrostatic enhancement mechanism or byternary complex formation. Se(IV) decreased Co(II) sorption at high Co(ll) surface loadings (>0.5 micromol/m2) where coprecipitation of Co(II) and A(III) in the form of layered double hydroxides (LDH) is expected to be the dominant sorption mechanism for the single-sorbate case. The extent of the Co(ll) sorption reduction in Co(III)/Se(IV) bisorbate systems compared to the corresponding single-sorbate systems increased with increasing Co(II) surface coverage. The rate of Co(II) desorption was reduced in the presence of Se(IV) compared to the single-sorbate case, indicating a direct interaction between Co(II) and Se(IV). A reaction between Co(II) and Se(IV) is further supported by an increase in Se(IV) sorption in the same bisorbate samples where Co(II) sorption is decreased. Thus, the macroscopic data indicates Se(IV) may be altering the mechanism of Co(II) sorption, potentially forming a ternary surface complex or different surface precipitate. PMID- 11944672 TI - Spectroscopic studies of the effects of selenate and selenite on cobalt sorption to gamma-Al2O3. AB - The fate and transport of toxic metal ions and radionuclides in the environment is often controlled by sorption reactions. The extent of sorption of divalent metal cations is controlled by a number of factors including the presence of cosorbing or complexing ligands. To study the impact of anion cosorption on metal cation sorption behavior, Co(II) sorption to gamma-Al2O3 in the presence of selenium oxyanions was investigated. To aid in the interpretation of macroscopic sorption results, X-ray absorption spectroscopy (XAS) experiments were conducted on single and bisorbate samples where the Co(II) surface coverage of the bisorbate sample was greater than or equal to the single sorbate sample. XAS data for single sorbate Co(II) samples were consistent with reported spectra of Co(II) Al(III) layered double hydroxides (LDHs), indicating coprecipitates are forming in these samples. Comparison of data from single and bisorbate samples showed a decrease in the number of nearest neighbor cobalt atoms in the presence of Se(IV), irrespective of the order of Se(IV) addition and no change in Co coordination in the presence of Se(VI). The extent of the decrease in cobalt second shell features between single and bisorbate samples with equal Co(II) coverage increased with an increase in the Se:Co surface coverage ratio. This trend suggests that the effect of Se(IV) on Co(II) sorption is a function of the Se(IV) surface coverage. At low ratio values, Co(II)-Al(III) LDH precipitates dominate. Increasing the Se:Co surface coverage ratio results in a progressive conversion of coprecipitate to an unknown, disordered Co(ll)/Se(IV) phase. Based on macroscopic data, this new phase could be an LDH with Se(IV) in the interlayer, an alternative precipitate such as a mixed metal Se(IV) hydrate, or a ternary complex. PMID- 11944673 TI - Atmospheric DMSO degradation in the gas phase: Cl-DMSO reaction. Temperature dependence and products. AB - The reactions of Cl atoms and ClO radicals with CH3-SOCH3 (DMSO) have been studied using the discharge flow method with direct detection of DMSO, CO, and products by mass spectrometry. The absolute rate constant at room temperature measured for reaction 1, (CH3)2SO + Cl --> products, was k(1) = (1.7 +/- 0.3) x 10(-11) cm3 molecule(-1) s(-1). For reaction 2, (CH3)2SO + ClO --> products, only an upper limit could be established, k(2) < or = 6 x 10(-14) cm3 molecule(-1) s( 1) Reaction 1 has been found to proceed through adduct formation and further decomposition involving the cleavage of the C-S bound. The pressure effect on the Cl-DMSO reaction from 0.5 to 3 Torr was negligible, and the temperature dependence in the range 273-335 K was also very slight. The results obtained are related to previous studies of sulfur compounds, and the atmospheric implications are also discussed in relation to the homogeneous sinks of DMSO. Tropospheric lifetimes of DMSO based on average Cl and ClO concentrations and the measured rate constants have been calculated showing that the contribution of reaction 1 must be of minor relevance in the marine boundary layer. Reaction 2 is so slow that it does not play any role within the atmospheric sulfur chemistry. PMID- 11944675 TI - Antarctic springtime depletion of atmospheric mercury. AB - Unlike other heavy metals that are inherently associated with atmospheric aerosols, mercury in ambient air exists predominantly in the gaseous elemental form. Because of its prolonged atmospheric residence time, elemental mercury vapor is distributed on a global scale. Recently, Canadian researchers have discovered that total gaseous mercury levels in the lower tropospheric boundary layer in the Canadian Arctic are often significantly depleted during the months after polar sunrise. A possible explanation may involve oxidation of elemental mercury, followed by adsorption and deposition of the oxidized form, leading to an increased input of atmospheric mercury into the Arctic ecosystem. Here we present the first continuous high-time-resolution measurements of total gaseous mercury in the Antarctic covering a 12-month period between January 2000 and January 2001 at the German Antarctic research station Neumayer (70 degrees 39' S, 8 degrees 15' W). We report that mercury depletion events also occur in the Antarctic after polar sunrise and compare our measurements with a data setfrom Alert, Nunavut, Canada. We also present indications that BrO radicals and ozone play a key role in the boundary-layer chemistry during springtime mercury depletion events in the Antarctic troposphere. PMID- 11944674 TI - Enhanced nitrate removal efficiency in wetland microcosms using an episediment layer for denitrification. AB - Microbial denitrification is the principal removal mechanism for nitrogen in treatment wetlands. For this study, flow-through wetland microcosms were designed to testwhether variations in the macroporous structure of the denitrification zone affected overall nitrate removal. In a sediment-only treatment, carbon as cattail (Typha latifolia) litter was mixed throughout a porous sediment matrix. A second treatment contained a distinct layer of loosely aggregated litter pieces placed atop the sediment matrix to form an episediment zone. Results showed that across nine conditions (NO3- influent = 7, 36, 65 mg N L(-1) and t(res) = 2.2, 2.9, 4.4, 7.7 d; n = 48) average denitrification was 33% greater (p < 0.0001) in the episediment treatment than in the sediment-only treatment. Both sediment-only and episediment treatments followed apparent reaction kinetics close to first order, with k(epi) = 0.21 d(-1) for the episediment treatment and k(sed) = 0.12 d(-1) for the sediment-only treatment. Analysis of vertical nitrate profile data using diffusive and turbulent mixing models indicated that denitrification occurred in a partially mixed episediment layer, as well as in an underlying sediment layer. Approximately 40% of the nitrate removal that occurred in the episediment treatment was estimated to occur in the episediments themselves. We conclude that enhancementof an episediment layer can increase denitrification in treatment wetlands which receive nitrate in overlying water. PMID- 11944676 TI - Dynamic oxidation of gaseous mercury in the Arctic troposphere at polar sunrise. AB - Gaseous elemental mercury (Hg0) is a globally distributed air toxin with a long atmospheric residence time. Any process that reduces its atmospheric lifetime increases its potential accumulation in the biosphere. Our data from Barrow, AK, at 71 degrees N show that rapid, photochemically driven oxidation of boundary layer Hg0 after polar sunrise, probably by reactive halogens, creates a rapidly depositing species of oxidized gaseous mercury in the remote Arctic troposphere at concentrations in excess of 900 pg m(-3). This mercury accumulates in the snowpack during polar spring at an accelerated rate in a form that is bioavailable to bacteria and is released with snowmelt during the summer emergence of the Arctic ecosystem. Evidence suggests that this is a recent phenomenon that may be occurring throughout the earth's polar regions. PMID- 11944677 TI - Rainbow trout (Oncorhynchus mykiss) can eliminate chiral organochlorine compounds enantioselectively. AB - Dietary accumulation of four chiral organochlorine compounds--alpha hexachlorocyclohexane (alpha-HCH), trans-chlordane, and chlorobiphenyls (CBs) 95 and 136-by immature rainbow trout (Oncorhynchus mykiss) was studied to determine if fish can accumulate and eliminate these compounds enantioselectively. Fish rapidly accumulated all four compounds from food spiked at micrograms per gram concentrations during a 40-d feeding period. Depuration half-lives were from 13 d for (+/-)-alpha-HCH to 375 d for (+/-)-CB 136. Fish preferentially eliminated (-) trans-chlordane and (+)-CB 136, with significant nonracemic residues observed after 20 d. These results are consistent with field measurements of these compounds in fish as well as known metabolic pathways. Enantiomeric fractions (EFs) for these two compounds changed significantly over the course of the experiment, suggesting that trout were enantioselectively biotransforming the compounds during the 238-d depuration phase. CB 95 and alpha-HCH residues were racemic throughout the experiment. High biomagnification factors for CB 95 suggest that it was not metabolized. Minimum values for metabolic elimination rates calculated from EF suggest that at least 58% of the trans-chlordane depuration rate can be attributed to metabolism, and all of the CB 136 depuration rate can be attributed to it. This study highlights the potential of chiral analysis as a tracer of in vivo biotransformation processes of xenobiotic compounds. PMID- 11944678 TI - FTIR kinetic, product, and modeling study of the OH-initiated oxidation of 1 butanol in air. AB - A kinetic and product study was performed on the reaction of OH radicals with 1 butanol in a 480 L indoor photoreactor and also in the EUPHORE outdoor smog chamber in Valencia, Spain. Long path in situ FTIR spectroscopy and gas chromatography with photoionization detection were used to analyze reactants and products. Using a kinetic relative rate technique, a rate coefficient of k(OH + 1 butanol) = (8.28 +/- 0.85) x 10(-12) cm3 s(-1) was measured in 740 Torr synthetic air at 298 +/- 2 K. The reaction products observed and their fractional molar yields were (in percent) butanal (51.8 +/- 7.1), propanal (23.4 +/- 3.5), ethanal (12.7 +/- 2.2), and formaldehyde (43.4 +/- 2.4). In addition, the results support the probable formation of 4-hydroxy-2-butanone. Propanal, ethanal, and formaldehyde could also be formed in secondary reactions of some of the primary aldehydic products. However, under the conditions employed in the experiments, the contribution from secondary reactions is very minor. On the basis of the product studies, a detailed atmospheric degradation mechanism was constructed and tested against experimental data by chemical box model calculations. Measured and simulated concentration-time profiles for selected reactants were in excellent agreement. PMID- 11944679 TI - Tracing the history of nuclear releases: determination of 129I in tree rings. AB - Concentrations of the long-lived radioisotope 1291 were measured in dated tree rings in order to determine whether the distribution of this isotope reflects the history of nuclear deposition. 129I concentrations and 129I/127I ratios were analyzed in tree rings and bark samples from four trees at West Valley, NY, and from one tree at Rochester, NY. West Valley was the site of short-lived nuclear fuel reprocessing activities (1966-1972), while Rochester, located 115 km to the northeast, provided a regional control site for the study. The selected trees reflect different modes of fluid and nutrient transport in trees, with three species of ring-porous trees (elm, oak, and locust), one semidiffuse (cherry), and one diffuse-porous tree (maple). The results show that 1291 levels in ring porous trees, in which xylem or hydrologic tissue is localized in the outermost growth ring, are generally well correlated with the expected 1291 deposition pattern for the region. In contrast, tree rings of the more common semidiffuse to diffuse-porous wood, where xylem is disseminated throughout the trunk, show a less well developed 129I signal, probably due to the transport of iodine ions across annual rings. Iodine concentrations in the tree rings range from 0.04 to 2 mg/kg, 129I/127I ratios from 6 x 10(-10) to 3.8 x 10(-6). Tree bark and the outermost rings show significantly higher 129I concentrations than the wood of the trunk. The 129I/127I ratios for bark are very similar to values obtained for surface soil and water at the two localities, while inner rings have ratios similar to those in deeper layers of the soil, reflecting different pathways for 129I uptake and the differences in ambient 1291 levels between the atmosphere and deep soil. Although ring porous trees preserve the depositional pattern of nuclear releases, rings older than or close to the onset of the nuclear age have 129I/127I ratios significantly above the preanthropogenic level, suggesting that even in these trees some redistribution of 129I occurs throughout the trunk. Our results indicate that growth rings from ring-porous wood are useful in time series analyses of regional 129I deposition, yielding reliable information on relative changes in 129I concentrations but requiring caution in the reconstruction of absolute ambient concentrations during any given time. PMID- 11944680 TI - Catalysis of PAH biodegradation by humic acid shown in synchrotron infrared studies. AB - The role of humic acid (HA) in the biodegradation of toxic polycyclic aromatic hydrocarbons (PAHs) has been the subject of controversy, particularly in unsaturated environments. By utilizing an infrared spectromicroscope and a very bright, nondestructive synchrotron photon source, we monitored in situ and, over time, the influence of HA on the progression of degradation of pyrene (a model PAH) by a bacterial colony on a magnetite surface. Our results indicate that HA dramatically shortens the onset time for PAH biodegradation from 168 to 2 h. In the absence of HA, it takes the bacteria about 168 h to produce sufficient glycolipids to solubilize pyrene and make it bioavailable for biodegradation. These results will have large implications for the bioremediation of contaminated soils. PMID- 11944684 TI - Emissions from ships in the northwestern United States. AB - Recent inventory efforts have focused on developing nonroad inventories for emissions modeling and policy insights. Characterizing these inventories geographically and explicitly treating the uncertaintiesthat result from limited emissions testing, incomplete activity and usage data, and other important input parameters currently pose the largest methodological challenges. This paper presents a commercial marine vessel (CMV) emissions inventory for Washington and Oregon using detailed statistics regarding fuel consumption, vessel movements, and cargo volumes for the Columbia and Snake River systems. The inventory estimates emissions for oxides of nitrogen (NOx), particulate matter (PM), and oxides of sulfur (SOx). This analysis estimates that annual NOx emissions from marine transportation in the Columbia and Snake River systems in Washington and Oregon equal 6900 t of NOx (as NO2) per year, 2.6 times greater than previous NO, inventories for this region. Statewide CMV NO, emissions are estimated to be 9,800 t of NOx per year. By relying on a "bottom-up" fuel consumption model that includes vessel characteristics and transit information, the river system inventory may be more accurate than previous estimates. This inventory provides modelers with bounded parametric inputs for sensitivity analysis in pollution modeling. The ability to parametrically model the uncertainty in commercial marine vessel inventories also will help policy-makers determine whether better policy decisions can be enabled through further vessel testing and improved inventory resolution. PMID- 11944681 TI - Evaluation of gastrointestinal solubilization of petroleum hydrocarbon residues in soil using an in vitro physiologically based model. AB - Petroleum hydrocarbon residues in weathered soils may pose risks to humans through the ingestion pathway. To understand the factors controlling their gastrointestinal (GI) absorption, a newly developed experimental extraction protocol was used to model the GI solubility of total petroleum hydrocarbon (TPH) residues in highly weathered soils from different sites. The GI solubility of TPH residues was significantly higher for soil contaminated with diesel than with crude oil. Compared to the solubility of TPH residues during fasted state,the solubility of TPH residues during fat digestion was much greater. Diesel solubility increased from an average of 8% during the "gallbladder empty" phase of fasting (and less than 0.2% during the otherfasting phase) to an average of 16% during fat digestion. For crude oil, the solubility increased from an average of 1.2% during the gallbladder empty phase of fasting (and undetectable during the other fasting phase) to an average of 4.5% during fat digestion. Increasing the concentration of bile salts also increased GI solubility. GI solubility was reduced by soil organic carbon but enhanced by the TPH content. PMID- 11944683 TI - Interfacial tension of chlorinated aliphatic DNAPL mixtures as a function of organic phase composition. AB - This research evaluates the ability of three models to predict the organic liquid water interfacial tension (IFT) of chlorinated aliphatic hydrocarbon mixtures that are dense nonaqueous-phase liquids (DNAPLs). Prediction of the IFT is relevantto quantify processes such as DNAPL trapping in soil pores and kinetic interphase mass transfer. Three models are evaluated: the Fu et al. method (FU) [Fu, J.; Buqiang, L.; Zihao, W. Chem. Eng. Sci. 1986,41 (10), 2673-2679]; a modified version of the Apostoluk and Szymanowski method (AS) [Apostoluk, W.; Szymanowski, J. Solvent Extr. Ion Exch. 1996, 14 (4), 635-651], and a simple linear ideal mixing theory (LIMT). The FU and AS methods require knowledge of NAPL-phase mole fractions and mutual solubilities. The LIMT method requires the pure organic liquid IFT and DNAPL-phase mole fraction as model input. Forty chlorinated DNAPL mixtures were used. The mixtures include two-, three-, and four component DNAPL mixtures of tetrachloroethylene, trichloroethylene, 1,2-cis dichloroethylene, 1,2-trans-dichloroethylene, and carbon tetrachloride. Measured IFTvaries nearlylinearlywith DNAPL-phase mole fraction for the all the DNAPL mixtures except those that include 1,2-DCE. The FU and LIMT models generally provided acceptable results for all mixtures. The FU model yielded an average relative error in the predicted IFT of 6.4%, while the LIMT model exhibited an average error of 9.3%. The AS method exhibited an average error of 16.4%. PMID- 11944682 TI - Adsorption of Pb(ll) and Eu(III) by oxide minerals in the presence of natural and synthetic hydroxamate siderophores. AB - Trihydroxamate siderophores have been proposed for use as mediators of actinide and heavy metal mobility in contaminated subsurface zones. These microbially produced ligands, common in terrestrial and marine environments, recently have been derivatized synthetically to enhance their affinity for transuranic metal cations. However, the interactions between these synthetic derivative and adsorbed trace metals have not been characterized. In this paper we compare a natural siderophore, desferrioxamine-B (DFO-B), with its actinide-specific catecholate derivative, N-(2,3-dihydroxy-4-(methylamido)benzoyl)desferrioxamine-B (DFOMTA), as to their effect on the adsorption of Pb(II) and Eu(III) by goethite and boehmite. In the presence of 240 microM DFO-B, a strongly depleting effect on Eu(III) adsorption by goethite and boehmite occurred above pH 6. By contrast, almost total removal of Eu(III) from solution in the neutral to slightly acidic pH range was observed in the presence of either 10 or 100 microM DFOMTA, due primarilyto the formation of metal-DFOMTA precipitates. Addition of DFOMTA caused an increase in Pb(II) adsorption by goethite below pH 5, but a decrease above pH 5, such that the Pb(II) adsorption edge in the presence of DFOMTA strongly resembled the DFOMTA adsorption envelope, which showed a maximum near pH 5 and decreasing adsorption toward lower and higher pH. PMID- 11944685 TI - Rapid dating of recent aquatic sediments using Pu activities and 240Pu/239Pu as determined by quadrupole inductively coupled plasma mass spectrometry. AB - Quadrupole inductively coupled plasma mass spectrometry has been used to rapidly establish the chronology of recent aquatic sediments via measurements of the activities of 239Pu, 240Pu, and the atom ratio 240Pu/239Pu. Following addition of 0.007 Bq of a 242Pu spike isotope, Pu is leached from 3-20 g aliquots of dry ashed sediments with HNO3. A selective anion exchanger is used to preconcentrate Pu into approximately 2 mL aliquots, which are directly analyzed using a pneumatic nebulizer and double-pass spraychamber operating at 60 microL/min solution uptake rate. The ICPMS data collection is performed for 10 min per sample. The U concentrations were 0.01-0.05 microg/L in the analyzed solutions, and the interference of 238U1H+ upon 239Pu+ was negligible. The method has been applied to determining Pu activities, inventory, and 240Pu/239Pu in a complete sediment core from Old Woman Creek (Huron, OH). The Pu activity profiles, obtained in approximately 6 h of instrumental measurement time, are in agreement with a y spectrometric 137Cs profile. Peak 239+240Pu and 137Cs activities in the core were 1.60 +/- 0.02 and 47.8 +/- 0.8 Bq/kg, respectively; inventories were 108 +/- 2 Bq/m2 239+240Pu and 2710 +/- 40 Bq/m2 137Cs. Detection limits, based upon the analysis of 20 g samples, were 0.004 Bq/kg 239Pu, 0.012 Bq/kg 240Pu, and 0.012 Bq/kg 239+240Pu. 240Pu/239Pu atom ratios of 0.16-0.19 were obtained for all core intervals containing detectable Pu, which indicates that global fallout is the source of these radionuclides. PMID- 11944686 TI - Coupling effects during steady-state solute diffusion through a semipermeable clay membrane. AB - Two separate coupling effects are evaluated with respect to steady-state potassium chloride (KCl) diffusion through a bentonite-based geosynthetic clay liner (GCL) that behaves as a semipermeable membrane. Both of the coupling effects are correlated with measured chemico-osmotic efficiency coefficients, omega, that range from 0.14 to 0.63 for the GCL. The first coupling effect is an explicit (theoretical) salt-sieving effect expressed as a coupled effective salt diffusion coefficient, Domega*, that is lower than the true (uncoupled) effective salt diffusion coefficient, Ds*, because of the observed membrane behavior. However, the maximum difference between Domega* and Ds* based on measured chloride concentrations is relatively small (i.e., = 10%), and the difference decreases with decreasing omega (i.e., Domega* --> Ds* as omega --> 0). The second coupling effect is implicit (empirical) and is characterized by the measurement of concentration-dependent effective salt diffusion coefficients that results in an degrees 300% decrease in Ds* as omega increases from 0.14 to 0.63. The decrease in Ds* resulting from implicit coupling is attributed to solute exclusion described in terms of a restrictive tortuosity factor. PMID- 11944687 TI - Formation of PCDDs, PCDFs, and coplanar PCBs from polyvinyl chloride during combustion in an incinerator. AB - Exhaust gases from the combustion of poly(vinyl chloride) (PVC), polyethylene (PE), polystyrene (PS), poly(ethylene terephthalate) (PET), and theirvarious mixtureswere analyzed for PCDDs, PCDFs, and coplanar PCBs by gas chromatography/mass spectrometry (GC/MS) in order to investigate the role of PVC in these chlorinated compounds. Total amounts of dioxins (PCDDs + PCDFs) found in the samples were 11.7 ng/g PE alone, 1.17 ng/g from PS alone, 25.3 ng/g from PET alone, 448 ng/g from PE with PVC, 140 ng/g from PS with PVC, 126 ng/g from PET with PVC, 824 ng/g from PVC alone under low-CO level, and 8,920 ng/g from PVC alone under high-CO level. CO level in high-CO level condition was 880 ppm which was 20 times greater than that in low-CO level condition. Formation of coplanar PCBs ranged from 0.095 ng/g (PE alone) to 77 ng/g (PVC alone under high-CO level). There is a clear correlation between dioxin formation and chloride content. PCDFs composed 80% (PET + PVC)--98% (PET alone) of the total dioxins formed in the exhaust gases. The results indicate that PVC contributes significantly to the formation of PCDDs, PCDFs, and coplanar PCBs from mixtures of plastics upon combustion. PMID- 11944689 TI - Selective removal of cobalt species using nanofiltration membranes. AB - Selective removal of cobalt species from simulated nuclear liquid waste was investigated with different nanofiltration (NF) membranes at various solution pH levels, initial cobalt concentrations, and background ion concentrations. This study provides insight into the understanding of the relationships between rejections of a target compound (cobalt) and chemical equilibria of various species in the feed solution during NF. Particularly, the ratio of electrostatic rejection to steric rejection for different membranes used was quantitatively evaluated to find outthe relative significance in NF. Substantial cobalt rejection by NF was achieved along with partial separation of monovalent ionic species, although it depended on the level of liquid pH and the presence of background species. Greater cobalt rejection at increased pH was attributed to the precipitation of CoCO3(s) associated with natural carbonates originating from atmospheric CO2 gas rather than that of Co(OH)2(s). A loose NF membrane (e.g., NTR7410) gave as high a rejection as other tighter ones due to the stronger influence of electrostatic rejection, particularly at low pH where no cobalt precipitation was occurring. The decrease of cobalt rejection with the addition of boric acid was found to occur due to the formation of complexes between cobalt and boric acid, which was verified by the analyses of solution turbidity and near infrared spectroscopy. PMID- 11944688 TI - Photocatalytic degradation of aqueous 4-chlorophenol by silica-immobilized polyoxometalates. AB - The degradation of 4-chlorophenol with near-UV light by silica-immobilized polyoxometalate (POM-in-SiO2) catalysts has been studied. The silica-immobilized Na6W7O24 (SW7), H4W1032 (SW10), H3PW12O40 (SPW12), and H6P2W18O62 (SP2W18) were prepared by means of the sol-gel hydrothermal technique through the hydrolysis of tetraethoxysilane in aqueous solution of the corresponding polyoxometalate, respectively. The degradation of 4-chlorophenol was monitored by measuring Cl- and CO2 concentrations and analyzing reaction intermediates by GC/MS analysis. During irradiation, 4-chlorophenol first dechlorinated to form hydroquinone and p benzoquinone, and then these intermediates further mineralized to form CO2 and H2O. The degree to which 4-chlorophenol was mineralized by photocatalytic oxidation was investigated. Results indicate less than 15% for SW7 but nearly complete mineralization for SW10 after 60 min of photoirradiation. The present studies suggest that POM-in-SiO2 catalysts may be a novel type of photocatalyts for the purification of the environmentally chlorophenol-contaminated water. PMID- 11944690 TI - Dechlorination of lindane, dieldrin, tetrachloroethane, trichloroethene, and PVC in subcritical water. AB - Pure water has been used to dechlorinate aliphatic organics without the need for catalysts or other additives. Dehydrohalogenation (loss of HCI with the formation of a double bond) occurred at temperatures as low as 105-200 degrees C for 1,1,2,2-tetrachloroethane, lindane (1,2,3,4,5,6-hexachlorocyclohexane, gamma isomer), and dieldrin (1,2,3,4,10,10-hexachloro-6,7-epoxy-1,4,4a,5,6,7,8,8a octahydro-endo, exo-1,4:5,8-dimethanonaphthalene). Complete loss of the parent compounds was achieved in less than 1 h at 150, 200, and 300 degrees C for 1,1,2,2-tetrachloroethane, lindane, and dieldrin, respectively. The initial dechlorination of lindane had an activation energy of 84 kJ mol(-1) with an Arrhenius pre-exponential factor of 1.5 x 10(6) s(-1). Dehydrohalogenation of lindane formed trichlorobenzenes, followed by subsequent hydrolysis and hydride/chloride exchange to form chlorophenols, lower chlorobenzenes, and phenol as the major final product. Reaction of poly(vinyl chloride) at 300 degrees C for 1 h formed aromatic hydrocarbons ranging from benzene to anthracene and a char residue with a ca. 1:1 carbon-to-hydrogen ratio (mol/mol). The residue contained <1 wt % of chlorine compared to 57 wt % chlorine in the original polymer. All compounds tested yielded chloride ion as the major product (at higher temperatures), indicating that complete dechlorination of some aliphatic organochlorines may be feasible. PMID- 11944691 TI - Estimation of mechanochemical dechlorination rate of poly(vinyl chlorde). AB - Poly(vinyl chloride) (PVC) was ground in air with CaO in the presence of quartz powder as a grinding aid by a small-scale planetary ball mill to investigate the relation of the dechlorination rate of PVC with the impact energy of the balls calculated from a computer simulation based on the Discrete Element Method under various conditions. Mechanochemical dechlorination proceeds as the grinding progresses and is improved with an increase in both the mill speed and the amount of balls introduced into the mill. The same trend can be seen in the relation between the specific normal impact energy of the balls and the rotational speed. The relationship between the observed dechlorination rate and the computed normal impact energy of the balls is linear, with a correlation coefficient of 0.965. This relationship can be used to estimate the dechlorination rate of PVC in a large-scale planetary ball mill. PMID- 11944692 TI - Treatment of mine drainage using permeable reactive barrers: column experiments. AB - Permeable reactive barriers designed to enhance bacterial sulfate reduction and metal sulfide precipitation have the potential to prevent acid mine drainage and the associated release of dissolved metals. Two column experiments were conducted using simulated mine-drainage water to assess the performance of organic carbon based reactive mixtures under controlled groundwater flow conditions. The simulated mine drainage is typical of mine-drainage waterthat has undergone acid neutralization within aquifers. This water is near neutral in pH and contains elevated concentrations of Fe(II) and SO4. Minimum rates of SO4 removal averaged between 500 and 800 mmol d(-1) m(-3) over a 14-month period. Iron concentrations decreased from between 300 and 1200 mg/L in the influent to between <0.01 and 220 mg/L in the columns. Concentrations of Zn decreased from 0.6-1.2 mg/L in the input to between 0.01 and 0.15 mg/L in the effluent, and Ni concentrations decreased from between 0.8 and 12.8 mg/L to <0.01 mg/L. The pH increased slightly from typical input values of 5.5-6.0 to effluent values of 6.5-7.0. Alkalinity, generally <50 mg/L (as CaCO3) in the influent, increased to between 300 and 1,300 mg/L (as CaCO3) in the effluent from the columns. As a result of decreased Fe(II) concentrations and increased alkalinity, the acid-generating potential of the simulated mine-drainage water was removed, and a net acid-consuming potential was observed in the effluent water. PMID- 11944693 TI - Environmental remediation by an integrated microwave/ UV-illumination method. 1. Microwave-assisted degradation of rhodamine-B dye in aqueous TiO2 dispersions. AB - The photocatalytic decomposition of the cationic rhodamine-B (RhB) dye was examined in aqueous TiO2 dispersions using an integrated microwave/UV illumination (PD/MW) method. This procedure proved to be superior in the degradation of the dye than the TiO2 photocatalytic degradative method alone. With few exceptions, the integrated PD/MW method also proved superior for other chemical systems. The greater efficacy of the PD/MW technique appears to be the result of the following two considerations: (i) there is enhanced formation of reactive oxygen species (.OH radicals), as attested to by DMPO spin-trap ESR methods and their attack on the dye; and although speculative at this time, (ii) the activity of bulk water or the TiO2 particle surface is somehow affected by microwave radiation. The greater efficacy of the PD/MW degradation method was also observed at low concentrations of molecular oxygen and at low radiant excitance of the light source. A brief mechanistic description is given on the basis of results obtained on the two model compounds, (i) benzoic acid and (ii) pyronin-B dye. PMID- 11944694 TI - Efficient destruction of CF4 through in situ generation of alkali metals from heated alkali halide reducing mixtures. AB - Perfluorocarbons (PFCs) are the most potent green house gases that are very recalcitrant at destruction. An effective way of converting PFCs using hot solid reagents into safe products has been recently introduced. By investigating the thermal reductive destruction of tetrafluoromethane (CF4) we provided new insight and more physicochemical consideration on this novel process. The complete destruction of CF4was successfully achieved by flowing the gas through a heated reagent bed (400-950 degrees C) that contained powder mixtures of alkali halides, CaO, and Si. The silicon acted as a reducing agent of alkali halides for the in situ production of alkali metals, and the calcium oxide played the role of a halide ion acceptor. The absence of any single component in this ternary mixture drastically reduced the destruction efficiency of CF4. The CF4 destruction efficiencies with the solid reagent containing the alkali halide, MX, increased in the order of Li approximately Na < K < Cs for alkali cations and I < Br < Cl < F for halide anions. This trend agreed with the endothermicity of the alkali metal generation reaction: the higher the endothermicity, the lower the destruction efficiency. Alkali metal generation was indirectly detected by monitoring H2 production from its reaction with water. The production of alkali metals increased with NaF, KF, and CsF in this order. The CsF/CaO/Si system exhibited the complete destruction of CF4 at as low as 600 degrees C. The solid product analysis by X-ray diffraction (XRD) showed the formation of CaF2 and the depletion of Si with black carbon particles formed in the solid reagent residue. No CO/CO2 and toxic HF and SiF4 formation were detected in the exhaust gas. PMID- 11944695 TI - Green route for the utilization of chrome shavings (chromium-containing solid waste) in tanning industry. AB - Chromium-containing wastes from various industrial sectors are under critical review. Leather processing is one such industrial activity that generates chromium-bearing wastes in different forms. One of them is chrome shavings, and this contributes to an extent of 10% of the quantum of raw skins/hides processed, amounting to 0.8 million ton globally. In this study, the high protein content of chrome shavings has been utilized for reduction of chromium(VI) in the preparation of chrome tanning agent. This approach has been exploited for the development of two products: one with chrome shavings alone as reducing agent and the other with equal proportion of chrome shavings and molasses. The developed products exhibit more masking due to the formation of intermediate organic oligopeptides. This has been corroborated through the spectral, hydrolysis, and species-wise distribution studies. The formation of these organic masking agents helps in chrome tanning by shifting the precipitation point of chromium to relatively higher pH levels. Hence, the developed products find use as chrome tanning agents for leather processing, thus providing a means for better utilization of chrome shaving wastes. PMID- 11944696 TI - Chikungunya infection--an emerging disease in Malaysia. AB - Many countries neighboring Malaysia have reported human infections by chikungunya virus, a mosquito-borne togavirus belonging to the genus Alphavirus. However, although there is serological evidence of its presence in Malaysia, chikungunya virus has not been known to be associated with clinical illness in the country. An outbreak of chikungunya virus occurred in Klang, Malaysia, between December 1998 and February 1999. The majority of the cases were in adults and the clinical presentation was similar to classical chikungunya infections. Malaysia is heavily dependent on migrant workers from countries where chikungunya is endemic. It is speculated that the virus has been re-introduced into the country through the movement of these workers. PMID- 11944697 TI - Long-term effect of interferon therapy on incidence of cirrhosis and hepatocellular carcinoma in Thai patients with chronic hepatitis B. AB - The aim of this study was to assess the long-term effects of interferon (IFN) therapy on the incidence of disease progression to cirrhosis and hepatocellular carcinoma (HCC) in Thai patients with chronic hepatitis B. Sixty-seven patients with hepatitis B e antigen (HBeAg)-positive chronic hepatitis B who received IFN therapy were retrospectively analyzed. The average duration of follow-up was 59.4+/-30.9 months (ranging from 20 to 119 months). Seventy-two untreated patients with comparable clinical data and mean duration of follow-up served as a control group. During follow-up, 24 (35.8%) treated and 7 (9.7%) untreated patients had a sustained loss of HBeAg. However, none of the treated patients or controls became negative for hepatitis B s antigen (HBsAg). Among treated patients, the response was independent of type and dose of IFN, as well as the presence of steroid priming. In addition, 1 of 24 (4.2%) sustained responders and 6 of 43 (14%) non-responders progressed to cirrhosis whereas 16 of 72 (22.2%) in the control group progressed to such sequelae. The overall incidence of new cirrhosis in sustained responders was significantly lower than in the control group (p=0.04). HCC appeared in 11 cirrhotic patients: 9 (12.5%) in the control group and 2 (4.7%) of the non-responders, whereas none of the sustained responders developed HCC. The average period to detection of HCC was 70.5+/-12.4 months for non-responders and 65.3+/-27.6 months for the control group, with no significant differences between these groups. In conclusion, our data suggest that IFN therapy might prevent the progression of cirrhosis and the development of HCC in patients with chronic hepatitis B. These beneficial effects were particularly observed in those who achieved a sustained virological response to treatment. PMID- 11944698 TI - Hepatitis B virus seroprevalence and risk assessment among personnel of a governmental hospital in Bangkok. AB - At present, the risk for acquiring hepatitis B virus (HBV) among hospital personnel is high. A cross-sectional analytic study of 380 hospital personnel was conducted in a governmental hospital in Bangkok to investigate HBV sero prevalence and to assess risk factors in order to develop the risk assessment form for screening the occupational risk of HBV among this group. The studied personnel who had no histories of HBV vaccination and jaundice before working in the hospital were included by voluntary participation. All studied personnel were interviewed by using a structured questionnaire consisted of risk exposure factors and some medical histories. Blood specimens were collected for determining HBV sero-markers (HBsAg, Anti-HBs, and Anti-HBc) by an enzyme immunoassay. The risk factors were analyzed by using Odds ratio (OR), chi2-test, and multiple logistic regression. The results revealed that 48.68% were positive for any HBV markers. The HBsAg positive rate was 3.42%, anti-HBs +/- anti-HBc was 43.16 and 2.11% were positive only anti-HBc. The significant risk factors from univariate analysis were: age over 30 years (OR=3.15, p<0.0001), marital status (OR=2.19, p=0.0002), working in risk ward (OR=2.89, p=0.0274), duration of working over 5 years, (OR=2.81, p<0.0001), a history of accident from working (OR=1.58, p=0.0354), and a history of needle stick (OR=1.83, p=0.0064). After multivariate analysis, the significant risk factors included age over 30 years (OR=2.99, p<0.0001), sex: male (OR=3.05, p=0.0020), working in risk ward (OR=2.81, p=0.0337), and a history of needle stick (OR=2.16, p=0.0030). The risk assessment form was developed by using risk scores. The validity was calculated by the Receiving Operating Curve. The sensitivity of this form was approximately 50% and the specificity was 80% when the cut-off score at risk > or = 5 was used. PMID- 11944699 TI - Prevalence of congenital cytomegalovirus and Toxoplasma antibodies in Thailand. AB - Prevalence of CMV and toxoplasmosis antibodies were determined among normal newborn infants, suspected congenital neonates and pregnant women. Seropositive rates of CMV and toxoplasmosis were similar. Most of the pregnant women had CMV antibodies. The in-house ELISA for detection of Toxoplasma antibodies was developed and compared with the commercial kit with sensitivity and specificity of 90.47% and 96.74% respectively. PMID- 11944700 TI - Increase in the sensitivity of dengue diagnosis by combination of reverse transcriptase-polymerase chain reaction and passage on cell cultures. AB - We passaged 52 serum samples from dengue patients on C6/36 cells for 7 days and checked the culture fluids by RT-PCR. Two serum samples, which were negative by direct RT-PCR, became positive. One sample was collected on fever day 1 and the other on fever day 2. Results indicate that combination of reverse transcriptase polymerase chain reaction (RT-PCR) with passage of serum samples on C6/36 cells increases the sensitivity of dengue diagnosis. PMID- 11944701 TI - Comparative evaluation of various commercial assays for diagnosis of dengue fever. AB - Dengue fever (DF) is endemic in India and dengue hemorrhagic fever (DHF) has been reported with increasing frequency in the last decade. We evaluated three commercial assays for detection of antibodies to dengue virus, to assess their performance in a diagnostic laboratory. Sera from 58 patients collected during a febrile outbreak in New Delhi in 1997 were studied. The methods evaluated were MRL Diagnostic Dengue Fever Virus IgM Capture ELISA, Pan Bio Dengue Duo IgM and IgG Capture ELISA and Pan Bio Rapid Immunochromatographic test. The MRL ELISA correctly identified 97.8% (43 of 44) of samples as dengue positive while the Pan Bio Duo ELISA and Pan Bio RIT identified 95.45% (42 of 44). The sensitivities of both Pan Bio Duo ELISA and Pan Bio RIT for primary dengue and secondary dengue were 100% and 93.54% respectively. The specificity of three assays were MRL IgM ELISA 100%, Pan Bio Duo ELISA 92.8% and Pan Bio RIT 85.7%. PMID- 11944702 TI - Primary dengue infection: what are the clinical distinctions from secondary infection? AB - To determine the magnitude of the problem posed by primary dengue infection in children and the distinctive clinical clues that may differ from those with secondary infection, 996 children serologically diagnosed with dengue infection and admitted to the Department of Pediatrics, Chulalongkorn Hospital, Bangkok, Thailand between 1988 and 1995 were retrospectively reviewed. One hundred and thirty-nine cases (14.0%) were serologically proved to be primary dengue infection. Of these, 72 were males and 67 were females, with a mean age of 4.8 years. Common manifestations by order of frequency included fever (97.8%), hepatomegaly (71.9%), vomiting (59.0%), decreased appetite (55.4%), coryza (52.5%), drowsiness (39.6%), diarrhea (34.5%), rash (33.8%), abdominal pain (23.0%) and seizure (15.8%). The mean duration of fever before admission was 4.6 days. Common sites of bleeding were skin (41.7%), mucous membrane (14.4%) and the gastrointestinal tract (12.2%). Clinical diagnosis was categorized into dengue fever (22.3%), dengue hemorrhagic fever (60.4%) and dengue shock syndrome (17.3%). Three patients (2.2%) died. Compared with the children with secondary dengue infection (n=139), children with primary dengue infections tended to be younger, presented more commonly with coryza, diarrhea, rash and seizure; and less commonly with vomiting, headache and abdominal pain (p < 0.05). The maximal hematocrit level, the mean difference between maximal and minimal hematocrit values and the maximal percentage of neutrophils were significantly lower in the study group, whereas the maximal percentage of lymphocytes was significantly higher. Dengue fever was more common and dengue shock syndrome was less common in the study group (p < 0.05). This study has emphasized that primary dengue infection is not uncommon and is less severe than secondary infection. Clinical presentations and laboratory findings are somewhat different between the two conditions. PMID- 11944703 TI - Dengue hemorrhagic fever in infancy at Petchabun Hospital, Thailand. AB - Prospective study of dengue hemorrhagic fever in infancy at the Pediatric Department, Petchabun Hospital during May 1997 to April 1999 is reported. There were 31 patients (2.9%) from total DHF cases of 1,044. Ten patients from 31 (32.2%) were referred from district hospitals. Male to female ratio was 17:14 (1.2:1). Maximal, minimal and median ages were 11.5 months, 22 days and 8 months, respectively. All patients had signs, symptoms and laboratory results compatible with dengue hemorrhagic fever. Serologic response was primary dengue infection. Five patients (16.1%) had gastrointestinal hemorrhage, all of them needed blood transfusion. Crystalline solution was fluid of choice given to all patients. Three patients were complicated with febrile convulsions. All patients had rapid and uneventful recovery. PMID- 11944704 TI - Perinatal dengue infection. AB - We report a case of vertical transmission of dengue infection in an infant. The mother's was a term pregnancy with a history of chronic hypertension. She presented with high fever of 3 days duration 5 days prior to delivery. Her initial complete blood count showed platelet count of 64,000/mm3. Dengue hemorrhagic fever was diagnosed 2 days later and symptomatic treatment was given. During labor her platelets dropped to 11,000/mm3 and platelet concentrate was given. Cesarean section was performed due to prolonged second stage of labor. Her infant was normal at birth except for petechiae on the left thigh. The child's platelet count was 34,000/mm3 and low grade fever was detected on the first day. Clinical sepsis was suspected and antibiotic treatment was started and continued for 4 days until all the cultures came back as negative. Both mother and her baby made an uneventful recovery and were discharged 6 days after delivery with normal platelet counts. Maternal blood was positive for IgM antibody to dengue virus. Both cord blood and the baby's blood were positive for dengue virus serotype 2 by PCR. PMID- 11944705 TI - Prevalence of rotavirus diarrhea among outpatients and hospitalized patients: a comparison. AB - The prevalence of rotavirus diarrhea was compared in two settings, among children attending outpatient clinics and those hospitalized (inpatients) at Pune, India. A total of 489 and 628 fecal specimens were collected during October 1993 to September 1996 from outpatients and inpatients respectively. Overall occurrence of rotavirus diarrhea was more among hospitalized children. Using the stratification on the variable age, it is shown that age is indeed a confounding variable. The important finding of the study was, in < or = 6 months age group, it was observed that the occurrence of rotavirus diarrhea was more in the outpatients (30.26%) than among the inpatients (10.11%). Children of this age group are likely to be partially protected by maternal antibodies. The effect of seasonality and sex distribution did not differ in the two settings. It was found that G2 serotype was the major cause of diarrhea among the outpatients. PMID- 11944706 TI - Biochemical and hematological manifestations of HIV/AIDS in Chiang Mai, Thailand. AB - HIV/AIDS is a multifactorial and multi-step disease. No single treatment against AIDS can save a patient. Our last report showed that vitamin A, vitamin E and beta-carotene were decreased while malondialdehyde (MDA) was increased. This report aims to evaluate biochemical and hematological parameters in HIV/AIDS patients in Chiang Mai, Thailand by holistic approaches. Sera from HIV/AIDS patients were examined for sugar, cholesterol, uric acid, total protein, albumin, urea, creatinine, AST, ALT, ALP, total/direct bilirubin, vitamin E, MDA, total antioxidant capacity (TAC), beta-carotene, complete blood cell counts, platelet count, CD4 count, prothrombin time, partial prothrombin time and soluble Fas (sFas). The results found that sFas levels in sera prior to holistic approach was not different from reference values and not significantly correlate with CD4 and absolute lymphocyte count. sFas could not serve as putative marker for CD4 destruction. After 3 months CD4 count, MDA, vitamin E and TAC did not change statistically. This approach had no effect on liver and kidney functions, red blood cell, white blood cell, platelet counts, and blood clotting factors. This presentation may be some alternative approaches to combat HIV infections and AIDS, leading to stabilize or extend survival time which should further be elucidated. PMID- 11944707 TI - Does HIV infection accelerate the development of hepatocellular carcinoma? A case report in a young man. AB - Hepatocellular carcinoma (HCC) is an important cancer. It occurs more often in men than women, and occurs mostly in people 50 to 60 years old. HCC has not been previously reported in a young HIV-seropositive patient in Thailand. We documented a very rare case of HCC in a 33 year old man. He was diagnosed and treated as Salmonella septicemia and tuberculosis. However, additional diagnosis based on pathological study disclosed a moderately differentiated HCC. Immunohistochemical study of the liver tissue was positive for hepatitis B surface antigen (HBsAg). PMID- 11944708 TI - Diagnostic value of dot-enzyme-immunoassay test to detect outer membrane protein antigen in sera of patients with typhoid fever. AB - Typhoid fever is still an important public health problem in many developing countries especially in tropical parts of the world, as in Indonesia. This problem opens the way for a further study with the aim of finding an alternative serological test with a high degree of reliability for the detection of typhoid fever. Given the above mentioned purpose, a study on the reliability of a laboratory test, the dot-enzyme-immunoassay outer membrane protein (DOT-EIA-OMP) was conducted comprising sera from 207 subjects (44 adult typhoid patients, 43 adult nontyphoid patients and sera from 120 adult healthy individuals serving as controls. The result of the study revealed that the diagnostic sensitivity of the DOT-EIA-OMP test for the detection of typhoid fever can be classified as high (93.16%), the specificity as moderate (76.74%), the efficiency (accuracy), positive predictive value and negative predictive value as high (85.06%, 80.39% and 91.66% respectively). The within run and between days reproducibility of this test was very high (CV=0%). Analysis of data obtained indicated that the DOT-EIA OMP test was a reliable screening test for the establishment of the diagnosis of typhoid fever in health centers with simple laboratory facilities. The application of this test has to be more contemplated in countries where the cost of laboratory test is a problem. PMID- 11944709 TI - Etiology of acute lower respiratory tract infection in children at Srinagarind Hospital, Khon Kaen, Thailand. AB - We investigated the etiology of acute lower respiratory infection (ALRI) in children under 5 admitted to Srinagarind Hospital. The causative bacteria and viruses were determined by hemoculture and viral isolation from blood and nasopharyngeal aspirate samples. Antigens of respiratory syncytial virus (RSV) and Chlamydia trachomatis were detected using EIA. The 74 children less than 5 years of age with ALRI enrolled in our study were diagnosed with pneumonia (75.7%), croup (16.2%), and bronchiolitis (8.1%), respectively. Examination of blood or nasopharyngeal aspirate revealed viral or bacterial infections in 26 and 22 cases, respectively, whereas 5 of the children aged under 1 year (10%) were diagnosed with pneumonia caused by Chlamydia trachomatis. RSV was the most common virus detected (24.3%) and was associated with pneumonia and bronchiolitis, while the parainfluenza virus was the primary cause of croup. In cases of pneumonia, bacterial infections were identified in almost all of the cases: and Streptococcus pneumoniae and Haemophilus influenzae were the most commonly isolated (at 8.9% each). Mixed infections were detected in 8 cases (10.8%). The incidence of RSV infection peaked during the especially warm and cool seasons, whereas the bacterial infections were primarily associated with the relatively cool season. Our study indicates that a combined pneumococcal and Hib vaccine and a RSV vaccine would reduce the high rate of pneumonia in children under 5 years of age in Northeast Thailand. PMID- 11944710 TI - Seasonal distribution of enteropathogens detected from diarrheal stool and water samples collected in Kathmandu, Nepal. AB - A total of 334 diarrheal fecal samples (from 210 males and 124 females) collected in Kathmandu, Nepal, were studied for various kinds of enteropathogens. Overall, 33% (111/334) fecal samples were positive for one or more enteropathogens. There was no difference in detection rates between males and females. Enteropathogen detection rates in summer, winter, spring, and autumn were 61% (40/66), 52% (45/87), 31% (25/81), and 25% (25/100), respectively. Altogether eight species of bacteria, three genera of viruses, and five species of protozoan parasites were detected with considerable seasonal variations. Among the bacterial isolates, enteropathogenic Escherichia coli topped the list followed by Vibrio sp. Only one sample had Shigella (S. sonnei). Rotavirus type A was the most frequently detected among the enteric viruses, followed by human enterovirus and human adenovirus, respectively. Among the enteric protozoan parasites, Giardia intestinalis was the most frequently detected followed by Cryptosporidium parvum. Detection of bacterial and protozoan pathogens showed a slightly high tendency in the summer season compared with that in the other seasons (p>0.05), whereas the detection of viruses was significantly high in the winter season (p<0.05). Of the total 57 water samples, 43 (75%) showed one or more bacterial species out of which 51% (22/43) were E. coli. Among the E. coli isolates, 68% were EPEC. Enterohemorrhagic E. coli (O157) was not detected. PMID- 11944711 TI - Guillain-Barre syndrome and Campylobacter infection. AB - Campylobacter infection of the gastrointestinal tract has been observed as an antecedent illness in some patients with Guillain-Barre syndrome (GBS); these patients have been reported to have poor prognosis. We investigated 29 patients with GBS, admitted to our hospital from January 1996 to December 1999 for recent Campylobacter enteritis by culture of their stool specimens. Campylobacter upsaliensis and C. jejuni were isolated from stools of one patient each with acute motor axonal neuropathy (AMAN) and acute inflammatory demyelinating polyradiculoneuropathy (AIDP) respectively. The patient with C. upsaliensis infection was a 7 year-old male child who developed features of AMAN, 7 days after onset of diarrhea. He recovered gradually within 24 days with residual deficit in the form of foot drop. This deficit has persisted for last three and half years. The other patient with C. jejuni infection was a 9 year-old boy, who developed AIDP after 9 days of acute diarrhea. This patient recovered completely within 28 days of illness without any deficit. None of the patients had relapse of GBS. The present findings indicate the need of planned systematic studies to explore the role of C. upsaliensis and other campylobacters as agents of antecedent diarrhea in patients of GBS with different clinical presentations and prognosis. PMID- 11944712 TI - Blood culture confirmed typhoid fever in a provincial hospital in the Philippines. AB - Epidemiological features of blood culture confirmed typhoid fever and antibiotic sensitivity of isolated Salmonella typhi strains were investigated in Bohol Province in Central Philippines from where no earlier information of these was avialable. Typhoid fever is endemic elsewhere in the Philippines (eg Metropolitan Manila and surroundings) where also multidrug resistant S. typhi strains have been detected. A laboratory for surveillance of invasive bacterial infections was established in a tertiary care government hospital, in Bohol, Central Philippines, in 1994. Patients with suspected typhoid fever or other serious infection were managed and blood cultures from them were taken according to clinicians' judgment. Blood cultures were processed and the isolated bacteria identified using generally accepted methods. S. typhi and other Salmonella isolates were identified using commercial antisera. Patient data were collected from hospital records. Of a total of 4,699 blood cultures done during a period of 3 3/4 years, 1,530 (32%) were requested for suspected typhoid fever. S. typhi was the most common pathogen isolated from 422 patients (8.9%), followed by S. paratyphi A from 55 patients (1%). Most patients were young adults (43%) and school age children (28%). Male:female ratio was 1.5:1. Among the 422 patients, there were 9 (2%) deaths due to typhoid fever, all with complications. All S. typhi isolates were sensitive to chloramphenicol, cotrimoxazole, and ampicillin. Our observation on blood culture confirmed typhoid fever demonstrates its importance as a major infectious disease in Bohol and gives a sound basis for treatment of typhoid patients and for further clinical and epidemiologic studies of typhoid fever and for following antibiotic sensitivity of S. typhi in Bohol and elsewhere in the Philippines. PMID- 11944713 TI - Mycobacterium szulgai lymphadenitis mimicking Kikuchi's disease in Thailand. AB - Mycobacterium szulgai is rarely found to be a cause of reported infection. Two thirds of cases were reported as pulmonary presentations, while the rest were infections of soft tissues or bone. In Thailand, few pulmonary infections due to M. szulgai have been reported. This is the first case of M. szulgai lymphadenitis confirmed by tissue culture. The patient presented with prolonged fever and tender enlarged cervical nodes. Histological findings showed large histiocytes with necrotic background compatible with Kikuchi's disease. However, the culture proved the case to be one of M. szulgai infection. That means this mycobacterium should be included in the differential diagnosis of fever with lymphadenitis. PMID- 11944714 TI - Demonstration of the natural foci of tsutsugamushi disease in Nan Peng Lie Islands in China. AB - In recent years, the incidence of tsutsugamushi disease has increased in Nan Peng Lie Islands in China, and the disease has not been recorded in this region. The natural foci of tsutsugamushi disease were investigated in this paper. Isolation of Orientia tsutsugamushi and the study of preventive measures were also performed. The region was the island natural foci of south subtropical zone. The main host and vector were Rattus norvegicus and Leptotrombidium (L.) deliens respectively. The seasonal quantity trends of Rattus norvegicus and Leptotrombidium (L.) deliense were consistent with the incidence of human infection in the region. The strains of O. tsutsugamushi were isolated from Rattus norvegicus and Leptotrombidium (L.) deliense. The identification showed that most strains were Karp. The seroepidemiology showed a high prevalence of antibody against O. tsutsugamushi. After preventive measures were implemented, the incidence was descent. So Nan Peng Lie Islands were the natural foci of tsutsugamushi disease. PMID- 11944715 TI - EPI vaccination in Nepal. AB - A number of surveys, small or large, have been undertaken by various agencies for coverage evaluation of immunization programs. The most commonly used design is the WHO-30 cluster sampling method. Other new methods are the Institute for Refresh Medical Statistics (IRMS) New Delhi method and the lot quality assurance sampling method. This paper describes the National Immunization Day (NID) method to evaluate the immunization coverage of the Expanded Program on Immunization (EPI) vaccines in the Sunsari district of Nepal. A total number of 3,332 respondents (69.4% females and 30.6% males) were interviewed regarding the immunization status of their children during NID. The children with complete immunization (BCG, measles and three doses of DPT and OPV) were 65.7%. The individual coverage by EPI vaccines (except OPV III and measles) was more than 80%. The result shows that there is positive relationship between immunization coverage and educational level of the respondents. PMID- 11944716 TI - Distribution of unengorged larvae of Leptotrombidium pallidum and other species in and around the rodent nest holes. AB - The distribution of unengorged larvae of Leptotrombidium pallidum, L. fuji and L. kitasatoi in and around 12 rodent-nest holes in Oita Prefecture, Japan was studied using the Tullgren funnel apparatus. Soil was taken from each nest hole, and the ground-surface soil and litter from the surrounding area A (an inner quadrate of 20 cm x 20 cm except the nest hole), and also from the outer area B (an outer quadrate of 40 cm x 40 cm excluding A and the nest hole) were sampled, separately. The numbers (% of the total) of L. pallidum collected from soil samples of the nest holes and areas A and B were 38 (19.0), 111 (55.5) and 30 (15.0); those of L. fuji were 171 (58.8), 104 (35.7) and 14 (4.8); those of L. kitasatoi were 35 (77.9), 7 (15.6) and 3 (6.7), and those of G. saduski were 20 (50.0), 17 (42.5) and 3 (7.5). The larvae recovered from litter samples were few, representing 0-8.5% of the total. It is shown that unengorged larvae of these species are distributed not only in the nest holes but also in the nearby areas, and exist mainly on (or in) the soil. PMID- 11944717 TI - Study on detecting antibodies to Toxoplasma gondii in pooled serum of blood donors by Dot-IGSS. AB - To prevent post-transfusion toxoplasmosis, the sera of blood donors, per six of which were mixed as a pool, were examined for anti-Toxoplasma gondii antibiodies by Dot-immunogold silver staining (Dot-IGSS) with the single serum specimens examined simultaneously. The results showed that the sensitivity and specificity of serum pool method were 92.31% and 99.96% respectively. The consistent rate between the two methods was 99.73% and kappa value was 0.947 (p<0.01). Considering the mean infection rate of Toxoplasma gondii being 4.86% in China, if the serum pool method be adoped, with pool size k=5, a 57% reduction in the number of tests, as well as the cost of the screen, can be expected. Beside the social benefit, consequent upon the interruption of the Toxoplasma gondii infection spread through blood transfusion also can be expected. PMID- 11944718 TI - Production of monoclonal antibodies to the cuticle of advanced third-stage larva of Gnathostoma spinigerum. AB - This study has demonstrated that sera from Balb/c mice infected with live advanced third-stage larvae (aL3), but not those immunized with crude larval extract, immunoprecipitated the 25-kDa protein from surface-iodinated extract of aL3. Hybridoma cell lines derived from spleen cells of an infected mouse secreted antibodies that reacted with several tissue of aL3 including the esophagus, intestine, muscle and cuticle by immunofluorescence assay. However, none of the cuticle-positive hybridoma cell lines produced antibodies that recognized surface iodinated protein of aL3 by immunoprecipitation. Western blot analysis showed that monoclonal antibodies (MAbs) secreted by clones derived from one of the cuticle-positive hybridoma lines recognized proteins of molecular weights ranging from 55-96 kDa. The MAbs most likely reacted with the collagenous component of the cuticle. PMID- 11944719 TI - Schistosoma haematobium infections in two rural communities of Edo State, Nigeria. AB - An epidemiological study of 1,136 inhabitants from two rural communities in Owan East local government area of Edo State, Nigeria was investigated to ascertain the prevalence, intensities and urinary symptoms in Schistosoma haematobium infections. In both communities, 371 (32.6%) of the villagers screened, excreted S. haematobium with a mean of 40.1 ova per 10 ml of their urine. The pattern of infection was highest among the school children, moderate among the farmers and least among the civil servants. The sensitivities of their urinary symptoms associated with this parasitic infection in these communities are 78.7% hematuria, 71.9% proteinuria, 70.4% supra public pain/discomforts and 59.6% dysuria. These foci of infections will broaden the epidemiological picture of urinary schistosomiasis in this part of the globe. PMID- 11944720 TI - Gender variations in the prevalence of parasitic infections and the level of awareness in adolescents in rural Nepal. AB - The study was conducted in rural school adolescent children to investigate the awareness and its association in parasitic infections in boys and girls. Of the 182 children examined 119 (65.3%) were male and 63 (34.6%) were female, age ranged 12-20 years with median age 15 years. Out of 182 stool samples examined 73 (40%) were found to be positive for parasites in which two or more parasites were found in 10 stool samples. Giardia lamblia 33 (18.1%) was the predominant parasite followed by hook worm 27 (14.8%) Entamoeba histolytica 13 (7.1%), Ascaris lumbricoides 05 (2.7%), Hyminolepis nana 02 (2.2%) and Trichuris trichiuria 01 (0.5%). Thirty-one (49.2%) in 63 females and 40 (33.6%) of the 119 males were positive for parasitic infections. The prevalence of worm infection was significantly higher in female children than male (p < or = 0.05). In contrast to the high parasitic prevalence rate in females they possessed significantly higher levels of awareness about parasitic infections. Out of 119 males 99 (83.2%) and 61 (96.8%) of the 63 females (p < or = 0.05) knew that worms suck food from host body. Similarly, 62.2% of males and 96.85 of females (p < or = 0.05) knew that parasites suck blood from human body. More study should be carried out to find out the gender difference in parasitic infection and level of exposure to risk factors. PMID- 11944721 TI - Transmission of bancroftian filariasis in tea agro-ecosystem of Assam, India. AB - Tea industry is a labor intensive agro-industry and filariasis is mostly localized among the tea garden workers in Assam. The workers live inside the gardens in colonies. Studies conducted in two cosecutive years revealed that among the host seeking Culex quinquefasciatus average infection rate was 4.6% and with 2.1 larvae per mosquito. The overall prevalence of infective mosquitos was 0.8% with average L3 load of 2.0 per mosquito. The probability of infected mosquitos surviving to have complete development of filarial larvae (13 days) was 0.17. The expectation of infective life was 1.416 days for man biting Cx. quinquefasciatus and the estimated adult survival rate of was 87.6%. It has been estimated that a total of 22,569 mosquito bites were received/man/year in tea garden environment out of which 182 bites/man/year were infective (0.806%). The monthly biting rate varied from 310-4,758.5 bites per man (mean 1,846 +/- 1,389.7 SD). Monthly transmission index of W. bancrofti filaria showed two periods of transmission. In both the year no infection was detected during February and March and infection rate remained low up to May (average infection in April 0.72% and in May 0.48%). PMID- 11944722 TI - Brugia malayi in a naturally infected cat from Narathiwat Province, southern Thailand. AB - Brugia malayi-like from an infected cat from Narathiwat Province, southern Thailand was identified intensively by microfilarial morphometry, acid phosphatase activity, and adult morphology. The results indicated that both microfilarial and adult characteristics conformed to the topotypic B. malayi. PMID- 11944723 TI - Gender, mosquitos and malaria: implications for community development programs in Laputta, Myanmar. AB - This paper examines the gender roles linked to division of labor and potential exposure to mosquitos and malaria prevention activities. A "Human Development Initiative" (HDI) Project has been launched in Laputta, a mangrove delta region of Myanmar assisted by United Nations Development Program since 1994. The project aims to improve rural community access to primary health care and provide micro credit programs, income generation schemes, and educational opportunities as a basis for community empowerment. Women and children of low-income households are the target beneficiaries. Prior to self-care training program and distribution of self-care manuals, altogether 20 focus group discussions (separately assigned to men and women) were conducted in eight study villages between January to February 2000. The primary vector for malaria in study area is Anopheles sundaicus. Rural women were prone to malaria due to exposure to mosquitos within the peak biting period at night because of their gender assigned roles. Both men and women perceived that mosquitos commonly bite before midnight, more at dusk. Lack of awareness of correlation between mosquitos and malaria together with lack of affordability enhance either non-use or shared use of bed-nets at home. Rural women did not consider destruction of breeding places of mosquitos as their major concern. Thus, it is essential for program planners to motivate local women for more active participation in vector control measures within and beyond their households in the context of community development programs. PMID- 11944725 TI - Poverty and malaria: a study in a Thai-Myanmar border area. AB - An eight-month qualitative study was conducted in 1999 in four villages of Bong Tee subdistrict, Kanchanaburi Province located along the Thai-Myanmar border area using in-depth interviews of key informants and malaria survey as research methodologies. Malaria was a serious problem in 39.6% of the families surveyed in June 1999. The four villages located in a valley covered with forests and small streams which were ideal for malaria epidemic. The structure of the villages has been changed from stable communities to disrupted ones divided along ethnic and class lines. There were 5 ethnic groups dominated by ethnic Karen. Villagers were poor and thus deprived of anti-malaria resources which allow them to remain exposed to malaria. PMID- 11944724 TI - Malaria past and present: the case of North Sulawesi, Indonesia. AB - The incidence and impact of malaria in North Sulawesi have declined both in the short term during the 1990s, and over a much longer timespan (though perhaps less continuously) since the end of the colonial period. The improvement already seems to have been well underway before deliberate vector control activities became extensive in the second half of the 1970s, and environmental changes affecting the Anopheles mosquito fauna, in particular the replacement of primary and secondary forest by permanent farmland, are probably the principal reasons for the long-term trend; other possible factors include the increasing use of antimalarial drugs. The well-documented decline in malaria incidence over the years 1991-1997, nevertheless, probably reflects the unprecedented scale of residual insecticide spraying in the province during that period, while the slight resurgence of the disease in the last three years corresponds to the subsequent cessation of house spraying as a result of the current economic crisis. Despite the evident importance of environmental change as a factor ameliorating the malaria situation in the long term, experience from the colonial era suggests that the prospects for deliberate environmental management (species sanitation) as an alternative malaria control strategy are poor. PMID- 11944726 TI - A rapid polymerase chain reaction based method for identification of the Anopheles dirus sibling species. AB - A simple polymerase chain reaction (PCR) based method was developed to differentiate the Anopheles dirus, species A, B, C and D in Thailand using specific primers designed from species specific sequences. The PCR protocol was optimized to obtain products of 120 bp, 75 bp, 60 bp and 172 bp for species A, B, C and D, respectively. This method used a cocktail of four primer sets to identify these An. dirus sibling species. The method is very sensitive as only a small portion of mosquito was required allowing the rest of the mosquito to be used for other analyses. Specimens also kept for up to 14 years could be analyzed unambiguously from either larvae or adult. This method is advantageous over other PCR-based methods for identification of malaria vectors because it does not require any specific DNA extraction. A mosquito specimen was homogenized in 1x PCR buffer, then the supernatant directly used for PCR identification, allowing a large number of samples to be processed at the same time. It provides a simple and rapid practical method for screening An. dirus species, which is essential in malaria vector epidemiological studies in Southeast Asia. PMID- 11944728 TI - Short stature and truncal shortening in transfusion dependent thalassemia patients: results from a thalassemia center in Malaysia. AB - One of the major complications in patients with transfusion dependent thalassemia is growth impairment secondary to iron overload. We studied the growth status in 66 patients with beta-thalassemia major and HbE-beta thalassemia who were transfusion dependent, aged from 2 to 24 years, and 66 controls matched for sex and age. The prevalence of short stature in transfusion-dependent thalassemics was 54.5% compared to 4.5% in control group (p<0.001). Short stature was more prevalent in those above the age of 10 years in this study group (83.3% vs 16.7%). Transfusion dependent thalassemics with short stature were found to have significantly lower mean standing height standard deviation scores (SDS), sitting height SDS and subischial leg length SDS values (p<0.001). There was also a significant difference between the mean sitting height SDS and the mean subischial leg length SDS in our thalassemics with short stature, suggesting that the short stature was due to disproportionate truncal shortening. Serum ferritin levels were significantly higher in transfusion dependent thalassemics who were short compared to those who were of normal height (p = 0.002). However, the mean pre-transfusion hemoglobin levels did not differ significantly between patients with short stature and those with normal height (p = 0.216). The prevalence of short stature also did not differ significantly between those with beta thalassemia major and those with HbE-beta thalassemia (p = 0.32). This study highlighted the importance of providing optimal treatment in these patients, including monitoring of growth parameters and optimizing iron chelation therapy. PMID- 11944727 TI - Aedes albolateralis, a potential vector of nocturnally subperiodic Wuchereria bancrofti and dengue type 2 virus. AB - The susceptibility of Aedes albolateralis to nocturnally subperiodic Wuchereria bancrofti and dengue type 2 virus was investigated by using artificial membrane feeding and intrathoracic inoculation techniques, respectively. The results indicated that Ae. albolateralis was susceptible to nocturnally subperiodic W. bancrofti (susceptibility rate = 9.43%) and dengue type 2 virus (susceptibility rate = 100%), suggesting the potential vector of the two pathogens. PMID- 11944729 TI - Natural course of autoimmune thyroiditis in Thai children. AB - Forty-seven pediatric patients with autoimmune thyroiditis were followed for an average of 5.18+/-2.89 years. The diagnosis was based on a firm goiter and a positive test for antithyroid antibodies. Initially, 23 patients had euthyroidism, 11 overt hypothyroidism, 6 compensated hypothyroidism and 7 with low TSH. All patients had clinical euthyroidism, except two who had overt hypothyroidism. The thyroid function tests, the size of the thyroid gland and the thyroid antibodies were regularly evaluated. After the follow-up, 26 patients had untreated euthyroidism, 12 with overt hypothyroidism received eltroxin for maintenance of euthyroidism, while 4 had compensated hypothyroidism and 5 low TSH levels. All had clinical euthyroid. The thyroid size was reduced in 12 patients (26%) while 4 (9%) had normal-sized gland. The goiter size in 35 patients (74%) remained unchanged. The antithyroglobulin and antimicrosomal antibody titers fluctuated higher in patients with overt hypothyroidism. Eltroxin was given only to those having overt hypothyroidism with diminished goiter size in 8 patients (73%). PMID- 11944730 TI - The prevalence of, and factors related to, compliance with glove utilization among nurses in Hospital Universiti Sains Malaysia. AB - Increasing risk of HIV infections among health care workers has been a continuing concern. The study was designed to identify the compliance of glove utilization, and factors related to non-compliance. A sample of 150 staff nurses were recruited from the study population of 550 nurses in Hospital Universiti Sains Malaysia. Data were collected by using a structured self-administered questionnaires. The response rate was 98.4%. The study revealed a low compliance (13.5%) of glove utilization (for all 9 procedures), which varied among different procedures (27-97%). Younger nurses and those with shorter duration of working experience had better knowledge of Universal Precautions. Nurses in intensive care unit and operation theatre were better in both knowledge and compliance of glove utilization. The three commonest misconceptions were identified as "selective use of gloves for high risk groups and suspected cases", and "tendency to depend on HIV prevalence". Nurses reported practical problems including administrative and personal related such as "stock irregularity" (46%), "glove not available at the emergency sites" (44%), and "reduction of tactile sensation" (39%). It was concluded that poor knowledge and practical problems were possible responsible factors for the low compliance. A good training for nurses comprising principle and practice of Universal Precautions, updated knowledge of blood and body fluid borne infections and risk and its management, will probably improve the compliance. PMID- 11944731 TI - Folate and vitamin B12 levels in patients with oral lichen planus, stomatitis or glossitis. AB - Forty-one patients with oral lesions and symptoms were enrolled in the study. Their ages ranged from 16 to 79 years with a mean age of 48.5 years. They were divided into two groups. Group I consisted of 25 patients with oral lichen planus and group II consisted of 16 patients with stomatitis or glossitis. Their complete blood counts, hemoglobin typing, serum and red cell folate, and serum vitamin B12 levels were studied. The results revealed low red cell folate levels in 11 out of 25 patients (44%) in group I and 9 out of 16 patients (56%) in group II. The serum vitamin B12 levels were within normal range in both groups. They were defined as having folate deficiency (n = 10), folate deficient erythropoiesis (n = 3) and folate depletion (n = 7). None of them had anemia nor macrocytes. Therefore, folate levels should be investigated in patients with oral lesions and symptoms especially those with risk factors of age, poor nutrition or systemic diseases. When suspected, daily folic acid supplements should be given. PMID- 11944732 TI - Correlation between Na, K-ATPase activity and potassium and magnesium contents in skeletal muscle of renal stone patients. AB - Samples of external oblique muscles were surgically removed from 45 renal stone patients and analyzed for their K, Na and Mg content. The muscle samples were also measured for membrane Na, K-ATPase activity from the assay of its K+ dependent 3-0-methyl fluorescein phosphatase (K+-dependent 3-0-MFPase) activity. The results showed that the mean muscle contents +/- SEM of K, Na and Mg were 65.2 +/- 1.7 (range, 41.1 to 86.1), 45.5 +/- 2.0 (range, 23.5 to 73.2) and 6.3 +/ 1.0 (range, 4.1 to 8.5) micromol/g wet weight, respectively. The mean activity +/- SEM of the K+-dependent 3-0-MFPase or the Na, K-ATPase was calculated by subtracting the activity of the basal-form from that of the total-3-0-MFPase, which was 113 +/- 21 (range, 11 to 177) nmol/g wet weight/minute. The activity of the Na, K-ATPase showed a significant correlation with muscle K-content (r = 0.52, p<0.001) and Mg content (r = 0.45, p<0.002). Though the external oblique muscles of renal stone patients in our study, as compared to data from other sources, had a considerably low concentration of K and Mg, they exhibited a good correlation with membrane-Na, K-ATPase activity. Our results, therefore, support previous observations made by other investigators. PMID- 11944733 TI - A long-term study on the efficacy of a herbal plant, Orthosiphon grandiflorus, and sodium potassium citrate in renal calculi treatment. AB - The study was performed to compare the efficacy of a herbal plant, Orthosiphon grandiflorus (OG), and the drug sodium potassium citrate (SPC) in treatment of renal calculi. Forty-eight rural stone formers identified by ultrasonography were recruited and randomly assigned to two treatment groups (G1 and G2). For a period up to 18 months, subjects in G1 received 2 cups of OG tea daily, each tea cup made from an OG tea bag (contained 2.5 g dry wt), and G2 received 5-10 g of granular SPC in solution divided into three times a day. Once every 5 to 7 weeks, subjects were interviewed, given an additional drug supply, administered a kidney ultrasound and had spot urine samples collected for relevant biochemical analysis. From the recorded ultrasound images, rates of stone size reduction per year (ROSRPY) were calculated. The mean ROSRPY was 28.6+/-16.0% and 33.8+/-23.6% for G1 and G2, respectively. These two means were not significantly different. ROSRPY values of G1 and G2 were combined and divided into three levels: Level A (ROSRPY > mean + 0.5 SD), Level M (ROSRPY = mean +/- 0.5 SD) and Level B (ROSRPY < mean - 0.5 SD). Dissolution of stones was least in Level B which was related to higher excretions of Ca and uric acid in the urine. After treatment, 90% of the initial clinical symptoms (ie back pain, headaches and joint pain) were relieved. Fatigue and loss of appetite were observed in 26.3% of G2 subjects. Our study indicates that treatment of renal calculi with OG tea is an alternative means of management. Further investigation is needed to improve dissolution of stones with a low ROSRPY. PMID- 11944734 TI - Monitoring of lead exposure among mechanics in Bangkok. AB - Lead is an important heavy metal used in many industrial processes. It plays an important role in the pathogenesis of both acute and chronic toxicity. Mechanics have a high risk of benzene exposure but tend to be a forgotten occupational group in Thailand. In this study, monitoring of lead exposure by anodic stripping voltammetry (ASV) determination of blood lead levels was conducted for 69 subjects (29 control subjects and 40 mechanics). The mean blood lead level in the control group was 6.59 +/- 1.98 microg/dl. The mean blood lead level in the mechanics was 11.24 +/- 5.82 microg/dl. There was a significant difference between the two groups (p < 0.05). Based on this study, consideration of the prevention of exposure to lead among mechanics as a public health strategy is recommended. PMID- 11944735 TI - Reporting clinical trials: full access to all the data. PMID- 11944736 TI - Protecting the most vulnerable: home mechanical ventilation as a case study in disability and medical care: report from an NIH conference. AB - Patients requiring chronic mechanical ventilation represent a particularly vulnerable segment of the expanding population of individuals with chronic disabilities. Many of these individuals can live successfully at home, but face significant obstacles. Current policies in health care coverage, durable medical equipment coverage, eligibility for assisted living, and licensing of caregivers all restrict the abilities of these individuals to live in the community. Prolonged home mechanical ventilation was pioneered in France, where a current international "best practice" provides a model for developing comprehensive, cost effective care for these individuals. PMID- 11944737 TI - Time-related changes in motor performance of the upper extremity ipsilateral to the side of the lesion in stroke survivors. AB - OBJECTIVE: The objective of the present study was to examine time-related changes in motor performance of daily tasks of the upper extremity ipsilateral to the side of lesion in poststroke hemiparetic patients. METHODS: Nine patients after an acute uniliteral cerebrovascular accident and 10 age-matched healthy controls were studied. Functional motor abilities of the upper extremity ipsilateral to side of lesion were examined over a 4-month time course, using validated measurement tools (Jebsen test of hand function, nine-hole peg test, and three functional activities of daily living). RESULTS: The results indicated a significant impairment in the motor function of the hand ipsilateral to the side of brain lesion in comparison with the matched extremity in control subjects. They also pointed to time-related improvement in performance speed, implying that the deterioration in the functional performance of the upper extremity on the uninvolved body side of poststroke hemiparetic patients is not static and may improve with time. The findings further suggested that the left hand of patients with an intact right cerebral hemisphere improves more than does the right hand of their peers whose left cerebral hemisphere is intact. CONCLUSIONS: Due to the small sample size and methodologic considerations, further and more extensive work is required to determine difference in improvement in motor abilities of the ipsilateral left versus the ipsilateral right upper extremities in stroke survivors. PMID- 11944739 TI - Selegiline treatment facilitates recovery after stroke. AB - OBJECTIVE: Selegiline (L-deprenyl) is a selective monoamine oxidase B (MAO-B) inhibitor used in the treatment of Parkinson's disease. In addition, it is thought to rescue neurons with a loss of target-derived trophic support. Several mechanisms have been proposed to explain these phenomena, such as the production of neurotrophic actions through astrocyte activation, reduction of free radical production, or the presence of antiapoptotic properties. The aim of this study was to investigate whether the systemic administration of selegiline facilitates recovery after a cerebral infarction in humans. METHODS: This phase II study was randomized, double-blind, and placebo controlled. Selegiline, 5 mg, or matched placebo was given twice a day for 3 months. The drug therapy was started within 48 h after a hemispheric infarction in the territory of middle cerebral artery. There were 24 patients recruited. Twenty patients were followed up to 3 months or until their death, and they represent the efficacy analysis group. The primary efficacy parameters were Scandinavian Stroke Scale (SSS), Barthel Index (BI), and Fugl-Meyer Scale (FMS). Secondary parameters were Zung Self-Rating Depression Scale (ZDS) and 15-Dimensional Measure of Health Related Quality of Life test (15 D). RESULTS: SSS improved statistically significantly from the baseline when compared with placebo (p = 0.019). The results were parallel among the other two primary efficacy variables (BI and FMS), showing a positive trend for selegiline, although they did not reach statistical significance. Similarly, in the analysis of the secondary efficacy variables, both the 15-D test and ZDS supported this positive trend in favor of selegiline, although no statistically significant differences between groups were found (p = 0.06 in 15-D test). CONCLUSIONS: Selegiline seems to be beneficial after a cerebral infarction. This benefit may be due to the enhancement of the recovery process. PMID- 11944738 TI - Intraspinal implants of fibrin glue containing glial cell line-derived neurotrophic factor promote dorsal root regeneration into spinal cord. AB - OBJECTIVE: The purpose of this study was to determine whether glial cell line derived neurotrophic factor (GDNF) delivered intraspinally via a fibrin glue (FG) enhanced regeneration of cut dorsal root (DR). METHODS: FG containing GDNF was inserted into aspiration cavities in the lumbar enlargement of adult rats. The transected L5 DR stump was placed at the bottom of the cavity and sandwiched between the FG and the spinal cord. Regenerated DR axons were labeled with horseradish peroxidase (HRP) or with immunohistochemical methods for calcitonin gene-related peptide (CGRP). RESULTS: Primary afferent axons labeled with HRP regenerated into the spinal cord, received GDNF, and made frequent arborization there. Some of these were myelinated axons that established synapses on intraspinal neuronal profiles. CGRP-immunoreactive DR axons extended into the motor neurons and formed prominent varicosities around their cell bodies. Only a few axons regenerated into the spinal cords given FG without GDNF. CONCLUSIONS: Our results indicate that GDNF enhances regeneration of DR into the adult rat spinal cord and that GDNF may be effectively supplied to the intraspinal injury site via FG. Because the regenerated axons establish synapses on intraspinal neurons, this therapeutic strategy has the potential to help to rebuild spinal reflex circuits interrupted by spinal cord injury. PMID- 11944740 TI - Physiological responses to two wheelchair-racing exercise protocols. AB - OBJECTIVE: This study investigated the blood lactate accumulation during two wheelchair-racing protocols. During exercise, energy is required, which causes metabolism to increase and blood lactate to accumulate. Proper training can reduce the amount of blood lactate accumulation and increase tolerance to blood lactate accumulation during aerobic exercise. METHODS: Eleven male wheelchair elite track athletes with a spinal cord injury were tested to determine their blood lactate response to speed and resistance workouts. A computer-monitored wheelchair dynamometer was used during all exercise tests. The order of the test (e.g., speed or resistance workout) was randomized. Blood pressure and blood samples were taken immediately before and after completion of each exercise test. RESULTS: The difference between the blood lactate concentrations before and after testing were 8.56 +/- 4.42, and 11.67 +/- 4.69 mM for the resistance and speed tests, respectively. CONCLUSIONS: Our results showed that the blood lactate responses were significantly different for the resistance and speed workouts. PMID- 11944741 TI - Problems with interhemispheric transfer of information in complete or partial agenesis of the corpus callosum. AB - OBJECTIVE: We assessed the presence of poor interhemispheric communication in agenesis of the corpus callosum (ACC) and hydrocephalus. METHODS: With specially designed tests, nine children with ACC were investigated and compared with 11 controls to see the degree of impairment present. Two subjects with a stretched corpus callosum due to hydrocephalus also were tested. A subject with the corpus callosum divided was tested for comparison. RESULTS: Significant differences were found in tests of coordination and stereognosis both with ACC patients and the callosotomy subject. No impairments were found in the hydrocephalics. CONCLUSIONS: When compared with controls, ACC patients perform poorly in several tests. The callosotomy patient also showed evidence of impairment similar to that of the ACC patients. PMID- 11944742 TI - Physiotherapy approaches in the treatment of ataxic multiple sclerosis: a pilot study. AB - OBJECTIVE: This study was planned to investigate the efficacy of neuromuscular rehabilitation and Johnstone Pressure Splints in the patients who had ataxic multiple sclerosis. METHODS: Twenty-six outpatients with multiple sclerosis were the subjects of the study. The control group (n = 13) was given neuromuscular rehabilitation, whereas the study group (n = 13) was treated with Johnstone Pressure Splints in addition. RESULTS: In pre- and posttreatment data, significant differences were found in sensation, anterior balance, gait parameters, and Expanded Disability Status Scale (p < 0.05). An important difference was observed in walking-on-two-lines data within the groups (p < 0.05). There also was a statistically significant difference in pendular movements and dysdiadakokinesia (p < 0.05). When the posttreatment values were compared, there was no significant difference between sensation, anterior balance, gait parameters, equilibrium and nonequilibrium coordination tests, Expanded Disability Status Scale, cortical onset latency, and central conduction time of somatosensory evoked potentials and motor evoked potentials (p > 0.05). Comparison of values revealed an important difference in cortical onset-P37 peak amplitude of somatosensory evoked potentials (right limbs) in favor of the study group (p < 0.05). CONCLUSIONS: According to our study, it was determined that physiotherapy approaches were effective to decrease the ataxia. We conclude that the combination of suitable physiotherapy techniques is effective multiple sclerosis rehabilitation. PMID- 11944743 TI - Anosognosia for hemiplegia in stroke rehabilitation. AB - OBJECTIVE: The purpose of this study was to investigate anosognosia for hemiplegia (AHP) in the rehabilitation phase after onset of stroke. METHODS: Forty-six hemiplegic stroke patients, 29 with right hemisphere damage (RHD) and 17 with left hemisphere damage (LHD) were evaluated approximately 1 month after onset of stroke. Anosognosia was evaluated with an implicit measure designed to assess anosognosic behaviors (choosing between unimanual and bimanual tasks), in addition to a traditional explicit verbal measure. RESULTS: AHP was found in 28% of the RHD and 24% of the LHD group. The majority of patients with AHP in the RHD group had large lesions involving the frontal, parietal, or temporal lobes and had coexisting sensory deficits and unilateral spatial neglect, whereas the LHD patients with AHP had predominantly small subcortical lesions and no sensory or attentional deficits. The functional outcomes of AHP patients in both hemisphere groups revealed their inability to retain safety measures at discharge from rehabilitation (p < 0.036) and their need for assistance in basic and instrumental activities of daily living at follow-up. CONCLUSIONS: AHP presents a significant risk for negative functional outcome in stroke rehabilitation. The underlying mechanisms of AHP may be different for left and right hemisphere patients, therefore requiring different intervention approaches. PMID- 11944744 TI - Possible role of corticosteroids in nervous system plasticity: improvement in amblyopia after optic neuritis in the fellow eye treated with steroids. AB - OBJECTIVE: Amblyopia consists of reduced visual function in the absence of organic disease, caused by deficient visual stimulation, most commonly due to squint or refractive error. Amblyopia is thought to be reversible up until the age of approximately 8 years (critical period) and is usually treated with occlusion of the fellow eye. There is recent evidence for visual system plasticity extending beyond the critical period, supported by reports of improvement in visual acuity in the amblyopic eye after loss of vision in the contralateral eye. This suggests that the adult visual system exhibits sufficient plasticity to allow such improvement. We describe here improvement in visual acuity in three amblyopic patients after they received high-dose intravenous glucocorticoids for optic neuritis in the contralateral eye. METHODS: Clinical and neurological evaluation added. RESULTS: In all cases, the improvement was sustained, even after the recovery from the optic neuritis. CONCLUSIONS: Because steroids affect neural plasticity, we hypothesize that they facilitate and enhance visual improvement in amblyopia, a quality that may be tested in future controlled trials. PMID- 11944745 TI - Motor impairment as a predictor of functional recovery and guide to rehabilitation treatment after stroke. AB - OBJECTIVE: This study tests three hypotheses relevant for the efficient use of rehabilitation services after stroke: (a) the severity of initial motor impairment after stroke predicts discharge motor impairment and self-care mobility scores; (b) identification of those unlikely to show improvement in motor impairment can focus rehabilitzation efforts on use of compensatory techniques and assist devices; and (c) improvement in self-care mobility scores without change in motor impairment, balance, or cognition is a quantitative estimate of the value of teaching compensatory techniques and use of assist devices. METHODS: We studied 171 sequential patients previously independent in the community who were admitted for inpatient rehabilitation within 17 +/- 12 SD days of an initial, unilateral, hemispheric, ischemic stroke. Impairment was assessed using the Fugl-Meyer upper limb motor (ULM), lower limb motor (LLM), and upper plus lower limb total motor (TM) subscores. Disability was assessed using the Functional Independence Measure (FIM), FIM self-care (FIMS), FIM mobility (FIMM), and FIM self-care plus FIM mobility (FIMSM) subscores. Spearman correlation coefficients tested strength of association between dependent and independent variables, stepwise linear regression tested the effects of clinically relevant co-variables, and positive and negative predictive values (PPV, NPV) assessed the clinical relevance of outcome-prediction models. RESULTS: The highest correlations observed were between admission TM scores and the following discharge scores: TM (R = 0.92; p < 0.01), ULM (R = 0.91; p < 0.01), LLM (R = 0.82; p < 0.01), FIMSM (R = 0.67; p < 0.01), FIMM (R = 0.67; p < 0.001), FIM (R = 0.58; p < 0.0001). An admission TM score in the lowest quartile had a PPV of 0.74 for a discharge ULM score in the lowest quartile. An admission TM score in the highest quartile had a PPV of 0.86 for a discharge ULM score in the highest quartile. Similar but weaker PPVs were seen for admission TM scores and discharge LLM scores. Patients without significant change in TM scores (< or = 2 points) had a 17 +/- 9 SD improvement in FIMSM scores. CONCLUSIONS: Admission motor impairment scores (a) predict discharge impairment and activities of daily living mobility functional outcome; and (b) guide treatment toward improving motor impairment versus use of compensatory techniques and assistive devices. The use of compensatory techniques and assistive devices, without change in motor impairment, is associated with a 17 +/- 9 SD improvement in FIMSM score. PMID- 11944746 TI - Berg balance scale and outcome measures in acquired brain injury. AB - OBJECTIVE: To examine the relationship of the Berg Balance Scale (BBS) to outcome after acquired brain injury. METHODS: Forty consecutive patients with acquired brain injury were admitted for multidisciplinary rehabilitation. Patients were assessed with the BBS. The BBS was originally designed as a quantitative measure of balance and risk for falls in community-dwelling elderly patients. The BBS comprises 14 different tasks graded on a 56-point scale. Community-dwelling elders with a BBS score of < or = 42 have > 90% risk for falls. RESULTS: In our study, there were 27 patients with a low BBS score (< or = 42) and 13 patients with a high BBS score (> or = 43). The discharge total Functional Independence Measure (FIM) scores were lower in the low BBS patients (96.4 +/- 21.2) compared with the high BBS patients (111.5 +/- 12.5) (p < 0.007). The length of stay (LOS) was significantly longer in the low BBS patients (38.9 +/- 18.5 days) compared with the high BBS patients (14.2 +/- 6.1 days; p < 0.000). Among the three patients that experienced falls during their hospitalization, all exhibited low BBS scores. The admission BBS score strongly correlated with admission total FIM scores (r = 0.86; p < 0.000) and moderately correlated with discharge total FIM scores (r = 0.56; p < 0.000) and LOS (r = -0.55; p < 0.000). Using a multiple regression analysis, the admission FIM score was found to be the better predictor of discharge FIM scores, and time admitted after injury was the better predictor of LOS. CONCLUSIONS: Prediction of rehabilitative outcome might be enhanced by the use of the BBS scores in combination with other clinical measures on admission to inpatient acute rehabilitation. PMID- 11944747 TI - A walking stick for a pure akinesia patient. AB - We describe a 63-year-old female patient with pure akinesia whose gait was facilitated by a handmade converted walking stick. A posterior ventral pallidotomy had been performed, but it did not alleviate symptoms. Her husband made a walking stick with a wire loop at the bottom, perpendicular to the walking direction. When the patient stepped over the loop, the frozen gait was improved. This converted walking stick is easily made and inexpensive. Although the walking stick did not improve the patient's gait radically, use of the converted walking stick effectively improved the patient's daily life because successful treatment of pure akinesia cannot be established. PMID- 11944748 TI - Measuring subluxation of the hemiplegic shoulder: reliability of a method. AB - OBJECTIVE: Subluxation of the shoulder after stroke can be measured according to the method described by Van Langenberghe and Hogan. METHODS: To evaluate the reliability of this method, the shoulder radiographs of 25 patients were available for this study. Two independent raters each assessed these radiographs twice. RESULTS: The intrarater reliability was good: percentage of agreement was 88 and 84%, weighted kappa, 0.69 [95% confidence interval (CI), 0.38-1.0] and 0.78 (95% CI, 0.60-0.95) for raters 1 and 2, respectively. The interrater reliability was poor: percentage of agreement was 36 and 28%, kappa, 0.11 (95% CI, 0.0-0.31) and 0.09 (95% CI, 0.0-0.23) in sessions 1 and 2, respectively. Subsequently the original method was adjusted by combining two categories (no subluxation and beginning subluxation) into one ("no clinically important subluxation"). CONCLUSIONS: After this adjustment of the categories, the interrater reliability improved [percentage of agreement, 72%, and kappa, 0.49 (95% CI, 0.18-0.80)], but did not reach acceptable values. PMID- 11944749 TI - Romanow's interim report: sidestepping the federalist question. PMID- 11944750 TI - The most compelling lesson of September 11. PMID- 11944751 TI - Disordered eating attitudes among Canadian teenagers. PMID- 11944752 TI - Fibrinolytic therapy for acute ST-segment elevation myocardial infarction. PMID- 11944753 TI - Risk factors for cardiovascular disease. PMID- 11944755 TI - Bedside rationing. PMID- 11944754 TI - Waiting times for cancer surgery. PMID- 11944756 TI - Reporting the clinical importance of randomized controlled trials. PMID- 11944757 TI - Reporting the clinical importance of randomized controlled trials. PMID- 11944758 TI - Risk factors, hospital management and outcomes after acute myocardial infarction in South Asian Canadians and matched control subjects. AB - BACKGROUND: Coronary artery disease affects a significantly larger proportion of Canadians of South Asian origin than Canadians of other ethnic origins. We compared differences in presentation, risk factors and management of myocardial infarction (MI) between South Asian Canadians and matched control subjects. METHODS: We reviewed the charts of 553 South Asian patients and 553 non-South Asian matched control subjects presenting with acute MI (International Classification of Diseases code 410) to 2 hospitals in Canada from January 1994 to April 1999. We identified South Asian subjects by their surnames and first names, and by using self-reported ethnicity and country of birth when available. Patients of Southeast Asian and Middle Eastern origin were excluded. The remaining patients were classified as non-South Asian. Subjects were matched by age within 5 years, sex, discharge date within 6 months and hospital of admission. Presentation characteristics, risk factors and major complications were compared between the 2 groups. RESULTS: The median time from symptom onset to presentation with acute MI was significantly longer among the South Asian subjects than among the control subjects (3.92 v. 3.08 hours) (p = 0.04). The South Asians were more likely than the control subjects to have diabetes mellitus (43.4% v. 28.2%) (p < 0.001) despite having a significantly lower mean body mass index (25.7 v. 28.0) (p = 0.05) but were less likely to have hyperlipidemia (36.2% v. 42.7%, p = 0.05), to smoke (29.3% v. 67.8%) (p < 0.001) or to have pre existing vascular disease (49.4% v. 55.0%, p = 0.04). Treatment of acute MI was similar between the South Asian and matched control groups. Also similar were the in-hospital outcomes, including mortality (9.6% and 7.8%, p = 0.27). INTERPRETATION: There are clear differences in the risk factor profile between Canadians of South Asian origin and those of non-South Asian origin who have acute MI. Despite the higher incidence of cardiovascular disease in the South Asian population, our results indicate that the in-hospital case-fatality rate for MI is the same for South Asian and non-South Asian Canadians. PMID- 11944759 TI - Comparison of diagnostic decision rules and structured data collection in assessment of acute ankle injury. AB - BACKGROUND: Ankle decision rules help to determine which patients with ankle injuries should undergo radiography. However, these rules are limited by imperfect generalizability and sensitivity. The judgement of physicians, aided by structured data collection, is a potential alternative. We compared the diagnostic performance of 2 decision rules with the performance of physicians, aided by structured data collection, in ruling out fracture in patients with acute ankle injury. METHODS: Consecutive patients with acute ankle injury who visited the emergency department of a teaching community hospital in Amsterdam were included in the study. After taking the patient's history and performing a physical examination, the surgical resident in each case completed a specially developed structured data form incorporating all of the variables in the Ottawa and Leiden ankle rules, as well as some additional variables. The form then asked whether the resident thought radiography was necessary. Each patient then underwent ankle and midfoot radiography. The films were independently interpreted by a radiologist and a trauma surgeon, who were both blinded to the information on the data form. Sensitivity, specificity and the percentage of patients for whom radiography was recommended were the main outcome measures. RESULTS: Of 690 consecutive patients, 647 met the inclusion criteria. Fractures were observed in 74 (11%) of these patients. Sensitivity was 89% (95% confidence interval [CI] 80% to 95%) for the Ottawa ankle rules, 80% (95% CI 69% to 88%) for the Leiden ankle rule and 82% (95% CI 72% to 90%) for physicians' judgement. Specificity was 26% (95% CI 23% to 30%), 59% (95% CI 55% to 63%) and 68% (95% CI 64% to 71%) respectively. Radiography was recommended in 76% (95% CI 72% to 79%), 46% (95% CI 42% to 50%) and 38% (95% CI 34% to 42%) of cases respectively. The Ottawa rules missed 8 fractures, of which 1 was clinically significant, the Leiden rule missed 15 fractures, of which 5 were clinically significant, and the residents missed 13 fractures, of which 1 was clinically significant. INTERPRETATION: Physicians' judgement, aided by structured data collection, was similar to existing international and local decision rules in terms of sensitivity in identifying cases requiring radiography and may outperform these prediction rules in terms of minimizing radiographic examinations for patients with ankle trauma. PMID- 11944760 TI - Impact of reference-based pricing for angiotensin-converting enzyme inhibitors on drug utilization. AB - BACKGROUND: Increasing copayments for higher-priced prescription medications has been suggested as a means to help finance drug coverage for elderly patients, but evaluations of the impact of such policies are rare. The objective of this study was to analyze the effect of reference-based pricing of angiotensin-converting enzyme (ACE) inhibitors on drug utilization, cost savings and potential substitution with other medication classes. METHODS: We analyzed 36 months of claims data from British Columbia for 2 years before and 1 year after implementation of reference-based pricing (in January 1997). The 119,074 patients were community-living Pharmacare beneficiaries 65 years of age or older who used ACE inhibitors during the study period. The main outcomes were changes over time in use of ACE inhibitors, use of antihypertensive drugs and expenditures for antihypertensive drugs, as well as predictors of medication switching related to reference-based pricing. RESULTS: We observed a sharp decline (29%) in the use of higher-priced cost-shared ACE inhibitors immediately after implementation of the policy (p < 0.001). After a transition period, the post-implementation utilization rate for all ACE inhibitors was 11% lower than projected from pre implementation data. However, overall utilization of antihypertensives was unchanged (p = 0.40). The policy saved $6.7 million in pharmaceutical expenditures during its first 12 months. Patients with heart failure or diabetes mellitus who were taking a cost-shared ACE inhibitor were more likely to remain on the same medication after implementation of reference-based pricing (OR 1.12 [95% confidence interval, CI, 1.06-1.19] and 1.28 [95% CI 1.20-1.36] respectively). Patients with low-income status were more likely than those with high-income status to stop all antihypertensive therapy (OR 1.65 [95% CI 1.43 1.89]), which reflects a general trend toward discontinuation of therapy among these patients even before implementation of reference-based pricing. INTERPRETATION: Reference-based pricing in British Columbia achieved a sustained reduction in drug expenditures, and no changes in overall use of antihypertensive therapy were observed. Further research is needed on the overall health and economic effects of such policies. PMID- 11944761 TI - Listening to injured workers: how recovery expectations predict outcomes--a prospective study. AB - BACKGROUND: Rigorous evidence on factors affecting the prognosis of work-related soft-tissue injuries remains limited. Although shown to be important for a wide variety of clinical conditions, recovery expectations have rarely been assessed as prognostic factors for workers with soft-tissue injuries. We examined the predictive role of various measures of recovery expectations among workers with injuries resulting in time off work. METHODS: We identified a prospective cohort of 1566 injured workers shortly after they filed a claim for their injury with the Ontario Workers' Compensation Board (OWCB). They had soft-tissue injuries to the back or upper or lower extremities, had new, lost-time claims from May to November 1993 and were still off work at the time of the first interview. We interviewed participants by telephone within 3 weeks after the injury and measured their recovery expectations (perceptions regarding progress, expected change in condition, expected time until return to usual activities and expectations regarding return to usual job) along with other, potentially important prognostic factors. The primary outcome was total time receiving 100% wage-replacement benefits during the year following injury, obtained from OWCB administrative files. Self-reported measures of pain, health-related quality of life and functional status, obtained up to 4 times during the year following injury, were both independent predictors and secondary outcomes. RESULTS: The 4 measures of recovery expectations together explained one-sixth of the variation in time receiving benefits. All but expectations regarding return to usual job were individually predictive of time receiving benefits. Judging one's recovery as much better than expected resulted in a 30% (95% confidence interval [CI] 9% 46%) faster rate of stopping receiving benefits (and likely returning to work) compared with judging one's recovery as much worse than expected. Similarly, participants who expected to return to usual activities within 3 weeks had a 37% (95% CI 26%-47%) faster rate of stopping receiving benefits than those who responded "Don't know" to this question, and participants who stated that they were fully recovered or would get better soon had a 25% (CI 5%-40%) faster rate than those who thought they would never get or stay better. Positive recovery expectations were also associated with reductions in pain grade and improvement in functional status outcomes. INTERPRETATION: Expectations regarding recovery may provide useful information on the complex process of recovering from work related soft-tissue injuries. For clinicians, patients' negative or uncertain expectations may indicate the need for further probing and intervention on psychosocial factors to facilitate recovery. PMID- 11944763 TI - Why is calling an ACE an ACE so controversial? Evaluating reference-based pricing in British Columbia. PMID- 11944762 TI - Latent tuberculosis infection: old problem, new priorities. PMID- 11944765 TI - Collisions with wildlife: the rising toll. PMID- 11944764 TI - Clinical nutrition: 5. How much should Canadians eat? PMID- 11944766 TI - Reducing antibiotic use for acute bronchitis by giving patients written information. PMID- 11944767 TI - Herbal kava: reports of liver toxicity. PMID- 11944768 TI - Nail gun. PMID- 11944769 TI - The disease that transformed medicine: AIDS turns 20. PMID- 11944770 TI - Drug approvals taking too long? PMID- 11944771 TI - Australia eyes Canada's rural MDs. PMID- 11944773 TI - Neurologists warn about link between chiropractic, stroke. PMID- 11944772 TI - Pediatricians taking new look at corporal-punishment issue. PMID- 11944775 TI - Paramedics face dismissal for refusing flu shots. PMID- 11944776 TI - Fewer Canadians being prescribed antibiotics. PMID- 11944774 TI - Armed forces announce plan of attack on environmental hazards. PMID- 11944777 TI - Quebec moves toward full disclosure of medical errors. PMID- 11944778 TI - India cracks down on sex-determination tests. PMID- 11944779 TI - Peer review of clinical research applications: leveling the playing field. PMID- 11944780 TI - When the payer says "no!": ethical considerations in patient advocacy. AB - Spinal cord injury is a relatively infrequent but often catastrophic injury. As such, it presents special challenges for the individuals affected and the clinicians who care for them. Claims for reimbursement of acute care, rehabilitation, and long-term care are often denied, requiring patients and clinicians to exhaust every effort to ensure coverage for therapeutic interventions, preventive care, and durable medical equipment. Ethical dilemmas are presented within the framework of the US healthcare system, and practical strategies are discussed. PMID- 11944781 TI - Combined calcitriol-pamidronate therapy for bone hyperresorption in spinal cord injury. AB - OBJECTIVE: To determine the biochemical effects of combined calcitriol pamidronate therapy on bone hyperresorption in patients with spinal cord injury (SCI). METHODS: This was a retrospective study of 21 SCI inpatients (4 women and 17 men, mean age 34 years) treated for bone hyperresorption. Initial treatment was 0.5 microg oral calcitriol once daily and 1,250 mg CaCO3 twice a day (1000 mg elemental calcium/day). On days 4 through 6 following the initial treatment, patients received 30 mg pamidronate intravenously once daily (total of 3 doses). Urinary N-telopeptide (NTx) and calcium excretion rates, and serum parathyroid hormone (PTH), 25-hydroxyvitamin D (25-D), 1,25-dihydroxyvitamin D (1,25-D), calcium, and phosphorus levels were measured within 2 weeks prior to and 2 weeks following pamidronate therapy. RESULTS: Patients demonstrated increased urinary NTx and calcium excretion, indicative of bone hyperresorption, and suppressed PTH and 1,25-D levels as early as 9 days post-SCI. Combined calcitriol-pamidronate therapy decreased urinary NTx and calcium excretion by 71% (P < .001) and 73% (P < .001), respectively. This therapy also increased serum levels of PTH (P <.05) and 1,25-D (P < .005). Post-pamidronate hypocalcemia or hypophosphatemia was observed in 44% (P < .01) or 53% (P < .01), respectively. CONCLUSION: Combined calcitriol-pamidronate therapy significantly inhibited bone hyperresorption in SCI patients. PMID- 11944782 TI - Age-related differences in length of stays, hospitalization costs, and outcomes for an injury-matched sample of adults with paraplegia. AB - OBJECTIVE: To investigate the effects of age at injury on neurological and functional outcomes and hospitalization length of stays and charges following spinal cord injuries resulting in paraplegia. METHODS: Subjects were 180 adults with paraplegia who were assessed in acute care and inpatient rehabilitation as part of the National Institute on Disability and Rehabilitation Research Model Spinal Cord Injury Systems. Age differences were examined by separating the sample into 3 age groups (18-39, 40-59, and 60+ years). A matched block design was used to control for injury characteristics. Cramer's statistic was used to identify age-related differences in qualitative variables; 3 x 5 one-way analysis of variance identified the main effects of age on quantitative variables. Tukey post hoc tests were performed to identify differences between age and age x injury characteristic variable levels. OUTCOME AND TREATMENT MEASURES: American Spinal Injury Association motor index scores, Functional Independence Measure (FIM) motor scores, discharge to private residence ratios, and hospitalization length of stays and charges were outcome and treatment measures. RESULTS: Age related differences were found for etiology and health care plan, as well as for preinjury marital status, education level, and employment status. The main effects of age at injury were found for the following treatment and outcome measures: rehabilitation length of stays, FIM motor scores at rehabilitation discharge, FIM motor improvement (change), and FIM motor daily improvement (efficiency). Tukey post hoc tests revealed that older patients had longer rehabilitation stays, lower rehabilitation discharge FIM motor scores, and showed less improvement compared with younger and middle-aged injury-matched patients. No age-related differences were found in rates of discharge disposition. CONCLUSIONS: Using a matched block design procedure, older patients are discharged with lower levels of functional independence and show lower levels of improvement despite longer rehabilitation stays when compared with younger patients. Older patients' neurological recovery appears equivocal to younger patients' recovery. In contrast to findings with a matched tetraplegia sample, older and younger patients with paraplegia are discharged to private residences at similar rates. PMID- 11944783 TI - Renal cell carcinoma in patients with prior spinal cord injury. AB - INTRODUCTION: In patients with spinal cord injury (SCI), abdominal diseases such as renal carcinoma are often diagnosed and treated late in their course. METHODS: A population-based retrospective review of SCI patients receiving care for renal cell carcinoma (RCC) in all Department of Veterans Affairs (DVA) medical centers was conducted for fiscal years 1988 to 1998. RESULTS: Of 96 patients identified, 57 were evaluable and 27 met study criteria. The mean patient age was 59 (range, 41-79 years). The mean time between SCI and treatment for RCC was 25 years (range, 1-51 years). All patients were men; 22/27 (81%) had 1 or more comorbid conditions. RCC was an incidental finding on surveillance imaging studies in 81% (22/27) of the patients. All 27 patients were treated surgically, 74% (20/27) by radical nephrectomy and 26% (7/27) by partial nephrectomy. All tumors were renal cell adenocarcinomas. Pathological staging by the tumor, nodes, and metastasis system was possible in 25; 92% (23/25) of tumors were stage I and 8% (2/25) were stage II. Postoperative morbidity occurred in 56% (15/27), and death occurred in 7% (2/27). CONCLUSION: In SCI patients in the DVA system, diagnosis of RCC is usually the result of an incidental finding on surveillance imaging. Tumors are diagnosed at early stages and partial nephrectomy is often feasible. Many of the postoperative complications are related to the SCI, and may be preventable. PMID- 11944784 TI - Self-care independence after spinal cord injury: patient and therapist expectations and real life performance. AB - PURPOSE: To evaluate changes in self-care performance between rehabilitation discharge and 1 year follow-up and to compare patients' and therapists' expectations of self-care independence with actual self-care performance after discharge. METHODS: Prospective study of 55 patients with tetraplegia (21 complete and 34 incomplete) after spinal cord injury (SCI) consecutively admitted to a large urban rehabilitation hospital [injury levels: C4 (5), C5 (33), C6 (9), C7 (4), and C8 (4)]. Data were collected using structured patient interviews, occupational therapists' predictions of patients' self care, and Functional Independence Measure ratings by rehabilitation staff. RESULTS: Analysis showed that for all skills, expectations of patients (mainly African American men) were higher than therapists' predictions, and for 3 of these skills, the differences were statistically significant: eating (Z = -4.47, P < .001 ); bathing (Z = 3.67, P < .001 ); and dressing (Z = -3.16, P = .002). Occupational therapists were better predictors of patients' actual self-care performance at discharge than were patients, although both tended to overestimate patients' performance. There was no evidence that self-care capability declined between discharge and 12 months follow-up, but routine self-care performance varied widely across patient and personal circumstances. CONCLUSIONS: Differential attrition reduced the study sample to 25 SCI patients who were primarily more severely injured African American men. Results showed that these patients maintained their self-care performance 1 year after discharge, but more innovative strategies must be developed to customize self-care retraining to address the unique needs of this patient population. PMID- 11944785 TI - Carbohydrate and lipid metabolism in chronic spinal cord injury. AB - BACKGROUND: Abnormalities of carbohydrate and lipid metabolism are more common in the spinal cord injury (SCI) population than in the able-bodied population. This is an important consideration in the long-term care of individuals with SCI. DESIGN: Literature review. FINDINGS: When compared with the able-bodied population, people with SCI are more likely to have oral carbohydrate intolerance, insulin resistance, elevated low-density lipoprotein cholesterol, and reduced high-density lipoprotein cholesterol, associated with increased prevalences of diabetes mellitus and cardiovascular disease. CONCLUSIONS: Because of increased risk factors for diabetes mellitus and heart disease in individuals with SCI, modifiable risk factors should be addressed, eg, obesity, inactivity, dietary factors, and smoking. To reduce mortality and morbidity associated with these risk factors, periodic screening for carbohydrate and lipid abnormalities is recommended, with appropriate therapeutic interventions. PMID- 11944786 TI - Lower extremity compartment syndrome in an adolescent with spinal cord injury. AB - OBJECTIVE: Describe the unusual complication of lower extremity compartment syndrome occurring in an adolescent with spinal cord injury (SCI). METHODS: Case presentation. RESULTS: A 17-year-old male with C5 ASIA A complete SCI developed a compartment syndrome of his lower leg on the ninth day postinjury. Presenting signs included an equinus deformity of the foot, blackened induration over the anterior tibia, circumferential erythematous markings over the calf, large urticarial lesions over the knee, and calf swelling. The presumed etiology of the compartment syndrome was excessive pressure from elastic wraps, which were placed over gradient elastic stockings. Pressures were 51 mmHg in the superficial posterior, 50 mmHg in the deep posterior, 33 mmHg in the anterior, and 34 mmHg in the peroneal compartments. The patient also developed rhabdomyolysis with myoglobinuria. In addition to supportive care, the patient underwent a dual incision fasciotomy for compartment release. CONCLUSIONS: The development of lower extremity compartment syndrome was probably a result of excessive pressure applied by elastic wraps. Elastic wraps should be used with caution in individuals with SCI. PMID- 11944787 TI - Atelectasis and mucus plugging in spinal cord injury: case report and therapeutic approaches. AB - INTRODUCTION: The leading causes of morbidity and mortality in the spinal cord injury (SCI) population are airway mucus plugging and atelectasis. OBJECTIVE: To illustrate the risks of pulmonary disease in individuals with SCI, and present effective therapeutic interventions. DESIGN: Case study of a 60-year-old veteran with T7 ASIA A spinal cord injury, who presented with a complete collapse of the left lung. FINDINGS: This patient developed fever, sepsis, and acute renal failure following colonoscopy. Following nephrostomy to remove a calculus, chest x-ray revealed complete collapse of the left lung. Despite the severe degree of atelectasis, he exhibited only mild respiratory distress. Aggressive treatment including chest physiotherapy techniques and pharmacologic intervention (acetylcysteine; bronchodilators) resulted in significant radiographic and clinical improvement. After his return to the SCI unit, his respiratory function was monitored, and assisted cough techniques were continued. CONCLUSIONS: Individuals with SCI have high risk of pulmonary complications. Because of neurological deficits, the usual signs and symptoms may not be apparent. Optimal management depends upon awareness of the risks, and a thorough understanding of the pathophysiology of mucus plugging and atelectasis and the alterations in pulmonary mechanics (dependent on level of injury). PMID- 11944788 TI - Racism: perceptions of distress among African Americans. AB - Some scholars have suggested that stressful living conditions are a major source of mental disorder among African Americans (Krieger, 1999; Neighbors, 1990; Kessler & Neighbors, 1986). There has, however, been debate as to whether this higher level of distress is due to racism or the fact that African Americans are more often of lower socioeconomic status. Stressors that play a significant role in mental disorder might be expected to occur more frequently among African Americans than the general population. This paper attempts to provide empirical support for the notion that racism is a separate and unique source of stress for African Americans. Specifically, it was hypothesized that African Americans would report more experiences of (1) daily stress and (2) racism than other groups and (3) the impact of racial stress would be greater among African Americans. One hundred and fifty six participants completed the Daily Stress Inventory and the Experience of Discrimination questionnaire. Multivariate analysis of variance indicated that African Americans reported higher impact of discrimination scores than European Americans. There were no gender or ethnicity differences in daily stress or the number of racial incidents reported. The implications of the data are discussed. PMID- 11944789 TI - Beyond components: using fidelity scales to measure and assure choice in program implementation and quality assurance. AB - Fidelity scales have become an accepted part of intervention research. Initially, fidelity scales focused on critical components of an intervention. In this paper we argue that the next generation of fidelity scales should include key process variables such as choice. Since choice is an essential element in all empowerment and recovery driven intervention models, a fidelity scale for an enhanced version of the Individual Placement and Support (IPS) supported employment model that incorporates choice as a fundamental component was developed as part of a SAMHSA community action grant. The process for developing the choice component and the dimensions measured are also described. PMID- 11944790 TI - The consequences of official labels: an examination of the rights lost by the mentally ill and mentally incompetent ten years later. AB - This study presents a survey of state statutes which restrict the civil rights of persons with a mental illness or who have been declared mentally incompetent. Five civil rights (jury service, voting, holding public office, marriage, and parenting) are examined. The results of the 1999 study are compared with the results of a 1989 study, to reveal any trends in the restriction of the civil rights of those suffering from mental problems. This comparison reveals that states continue to restrict the rights of the mentally ill and incompetent, and that there is a trend toward increased restriction of the familial rights of marriage and parenting. PMID- 11944791 TI - Exploring mental health service needs for the elderly: results of a modified Delphi study. AB - Identified key informants with professional experience in the broad domain of mental health services reported current and future mental health service needs of elderly Harris County residents as part of a three-round modified Delphi study. Forty-one professionals from various housing, psychosocial, medical, and protective service provider groups were surveyed during three consecutive rounds. The consensus achieved through the Delphi process served to describe the gaps in the mental health services currently available to elderly Harris County residents as well as identify the service needs of this population into the future, thereby providing direction for future service development. PMID- 11944793 TI - Beneficence vs. obligation: challenges of the Americans with Disabilities Act for consumer employment in mental health services. AB - Involvement of mental health service consumers in the provision of mental health services is a growing model in community mental health. It is, however, a complicated issue, made ever more so by the passage of the Americans with Disabilities Act. In this ethnographic case study, we seek to explore the changes one social services agency has made to adjust to the requirements of the ADA and the impact of these changes on their consumer employees. Our results indicate potential for positive progress as a result of the ADA, but also unexpected pitfalls as organizational cultures change as well. PMID- 11944794 TI - A note on the emission of nitrogen oxides from silage in opened bunker silos. AB - The evolution of NOx from grass and maize silages was measured using chemiluminescence in samples kept in airtight containers, in the silos and in a 750 kg mass removed to a mixer waggon. Measurements were made on the grass and maize silos in two consecutive years. The results show that there is continuous evolution of NOx after silos have been opened and that high concentrations persist in the mass which are rapidly released on agitation at the feed-out. The maximum recorded concentrations of NO and NO2 were 1985 and 152 ppbv respectively. These values are orders of magnitude greater than for rural background levels and exceed the maximum hourly exposure of 50 ppbv for NO2 recommended by the UK expert panel for quality standards. PMID- 11944792 TI - Community mental health care for women with severe mental illness who are parents. AB - Women with SMI are involved in childbearing and childrearing, but may have problems with parenting related to symptoms, lack of knowledge and skills, or lack of environmental supports. They may need intensive services to manage their mental illness and to parent to the best of their abilities. Preliminary research suggests that mental health interventions can improve family functioning, but that these services are not widely available in community mental health systems. This paper describes services to help clients who are parents, which includes integration of adult's and children's services, long-term, home-based parent training, and linkage with community supports. PMID- 11944796 TI - The variability of estimates of variance, and its effect on power analysis in monitoring design. AB - Power analysis can be a valuable aid in the design of monitoring programs. It requires an estimate of variance, which may come from a pilot study or an existing study in a similar habitat. For marine benthic infauna, natural variation in abundances can be considerable, raising the question of reliability of variance estimates. We used two existing monitoring programs to generate multiple estimates of variance. These estimates were found to differ from nominated best estimates by 50% or more in 43% of cases, in turn leading to under or over-estimation of sample size in the design of a notional monitoring program. The two studies, from the same general area, using the same sampling methods and spanning a similar time scale, gave estimates varying by more than an order of magnitude for 25% of taxa. We suggest that pilot studies for ecological monitoring programs of marine infauna should include at least two sampling times. PMID- 11944795 TI - PAH levels and profiles in the suspended particulate matter in Zagreb through four seasons. AB - This paper presents the analysis of polycyclic aromatic hydrocarbons (PAHs) measured in all four seasons in suspended particulate matter (SPM) collected with a high-volume sampler on one measuring site in the northern part of Zagreb. About 30 samples of SPM were analysed for each season, including workdays and weekends and there were no differences amongst them. The concentrations of all PAHs were highest in winter and lowest in summer. The spring PAH concentrations were lower than the autumn ones, as the spring had more sunny and warm days. The profiles of PAH/BaP at the measurement sites showed that the main source of PAHs in spring and summer was traffic while a substantial amount of autumn and winter PAHs, besides traffic, came from heating. PMID- 11944797 TI - Radionuclides in soil and water near a low-level disposal site and potential ecological and human health impacts. AB - Material Disposal Area G is the primary low-level radioactive waste disposal site at Los Alamos National Laboratory, New Mexico, and is adjacent to Pueblo of San Ildefonso lands. Pueblo residents and Los Alamos scientists are concerned about radiological doses resulting from uptake of Area G radionuclides by mule deer (Odocoileus hemionus) and Rocky Mountain elk (Cervus elaphus), then consumption of deer and elk meat by humans. Tissue samples were collected from deer and elk accidentally killed near Area G and were analyzed for 3H, 90Sr, total U, 238Pu, 239,240Pu, 241Am, and 137Cs. These data were used to estimate human doses based on meat consumption of 23 kg y(-1). Human doses were also modeled using RESRAD, and dose rates to deer and elk were estimated with a screening model. Dose estimates to humans from tissue consumption were 2.9 x 10(-3) mSv y(-1) and 1.6 x 10(-3) mSv y(-1) from deer and elk, respectively, and RESRAD dose estimates were of the same order of magnitude. Estimated dose rates to deer and elk were 2.1 x 10(-4) mGy d(-1) and 4.7 x 10(-4) mGy d(-1), respectively. All estimated doses were significantly less than established exposure limits or guidelines. PMID- 11944799 TI - Monitoring of seasonal vegetables for pesticide residues. AB - Market samples (60) of six seasonal vegetables were monitored during 1996-1997 to determine the magnitude of pesticidal contamination. The estimation of insecticide residues representing four major chemical groups i.e. organochlorine, organophosphorous, synthetic pyrethroid and carbamate, was done by adopting a multiresidue analytical technique employing GC-ECD and GC-NPD systems with capillary columns. The tested samples showed 100% contamination with low but measurable amounts of residues. Among the four chemical groups, the organophosphates were dominant followed by organochlorines, synthetic pyrethroids and carbamates. About 23% of the samples showed contamination with organophosphorous compounds above their respective MRL values. More extensive studies covering different regions of Haryana state are suggested to get a clear idea of the magnitude of vegetable contamination with pesticide residues. PMID- 11944798 TI - Lead content of petrol and diesel and its assessment in an urban environment. AB - Pakistan is one of the few countries in Asia that continues to use only leaded petrol as vehicular fuel. The concentration of Pb in its petrol reported in 1991 was the highest (1.5-2.0 g Pb L(-1)) of all produced by the various Asian countries and far exceeded the WHO's guideline of 0.15 g Pb L(-1). We have undertaken a study to trace and quantify this toxic element in the environs of Karachi. Pakistan's major metropolis, having more than 30% of the nation's total number of vehicles. In this article we report the Pb contents of petrol and diesel currently manufactured and marketed in the city. Samples of 'Regular' petrol collected in 1999 was found to contain 0.363 g Pb L(-1) (range: 0.335 0.390 g Pb L(-1)), a factor of 5 lower than that marketed prior to 1991. Its concentration in diesel fuel was much lower (0.017 g Pb L(-1)). Based on the available statistical data on the type and volume of vehicular traffic, we assessed that the current lead emission from vehicular traffic into the atmosphere is 391 metric tons a year, which is a factor of 2.7 lower than that estimated for 1989. PMID- 11944800 TI - Macroinvertebrate regionalisation for use in the management of aquatic ecosystems in Victoria, Australia. AB - The development of a broader, more holistic approach to aquatic ecosystem management has been called for in recent years. Physical and chemical objectives alone are no longer considered sufficient for the protection of aquatic ecosystems and should be supplemented with biological objectives. The ubiquitous and sedentary nature of macroinvertebrates, combined with their measurable response to environmental conditions, favour their use as important indicators in environmental policies. To establish biological objectives, there is a need for a regional framework to limit the variability between ecosystems. Past studies have demonstrated that an a posteriori regionalisation approach may be more useful than an a priori approach in explaining single component (e.g. macroinvertebrates) patterns across ecosystems. This is particularly important as aquatic resource management agencies often focus on one or two components of the ecosystem to assess environmental health. This study uses an a posteriori method to delineate and describe biological regions based on edge and riffle macroinvertebrate data. The regionalisation will provide a framework for setting biological objectives, based on the range of reference conditions measured within each separate region. The objectives will include regional checklists for taxa and biotic indices. Predictive modelling in the style of RIVPACS or AUSRIVAS will also be used within each region to develop objectives, incorporating local, regional and systematic features as predictor variables. PMID- 11944801 TI - The accuracy of forest damage assessments--experiences from Sweden. AB - In this article the consistency of forest damage assessments on conifer trees is analysed, by using different methods in estimating the accuracy in assessments of defoliation and discolouration. The data originate from control surveys in the Swedish national forest damage survey, as well as from national and international training courses. Standard deviation of differences in the assessment of defoliation on single trees is found to be about 10% units for Norway spruce and 8.5% for Scots pine. Problems in correctly assessing damaged Scots pine trees and discoloured Norway spruce trees are revealed by measures of agreement (Kappa statistic). Results from several years of national training courses indicate that, on an average, the observer teams do not significantly differ from a national standard, but significant differences between observer teams are found. The presented estimates indicate a substantial within observer error compared to the between observer error. The results indicate that the long-term development of forest damage, rather than short-term fluctuations, is the most important information from these kinds of inventories. PMID- 11944802 TI - Immunological basis of differences in disease resistance in the chicken. AB - Genetic resistance to diseases is a multigenic trait governed mainly by the immune system and its interactions with many physiologic and environmental factors. In the adaptive immunity, T cell and B cell responses, the specific recognition of antigens and interactions between antigen presenting cells, T cells and B cells are crucial. It occurs through a network of mediator proteins such as the molecules of the major histocompatibility complex (MHC), T cell receptors, immunoglobulins and secreted proteins such as the cytokines and antibodies. The diversity of these proteins that mainly is due to an intrinsic polymorphism of the genes causes phenotypic variation in disease resistance. The well-known linkage of MHC polymorphism and Marek's disease resistance difference represents a classic model revealing immunological factors in resistance differences and diversity of mediator molecules. The molecular bases in any resistance variation to infectious pathogens are vaguely understood. This paper presents a review of the major immune mediators involved in resistance and susceptibility to infectious diseases and their functional mechanisms in the chicken. The genetic interaction of disease resistance with production traits and the environment is mentioned. PMID- 11944803 TI - Borna disease: current knowledge and virus detection in France. AB - For over two centuries, Borna disease (BD) has been described as a sporadically occurring infectious meningoencephalomyelitis affecting horses and sheep in Central Europe. Over the last decade, the BD epidemiology has been discussed. Firstly, its geographical distribution seems larger than what was previously thought. Secondly, the disease can affect a large number of warm-blooded animal species, including humans. The aetiological agent is the Boma disease virus (BDV), an enveloped, nonsegmented negative-stranded RNA virus classified in the new virus family Bornaviridae (Mononegavirales order). It can induce severe clinical signs of encephalitis with striking behavioural disturbances and may cause death. BDV genome has recently been detected in France in the blood and brain of several animal species (horses, bovines, foxes). PMID- 11944804 TI - Trophoblastic interferon-gamma: current knowledge and possible role(s) in early pig pregnancy. AB - In the pig as in ruminant species, the implantation of the elongated conceptus - the embryo with its associated membranes - onto the maternal uterus is accompanied by an intense secretion of interferon (IFN), which culminates at day 15 of development. It has been shown that in fact the pig trophectoderm - the polarized epithelium which lines the conceptus - simultaneously secretes two types of interferons: IFN-gamma (IFN-gamma), which is the more abundant species, is produced in very substantial amounts. Another IFN is also secreted, which happens to be a novel type I IFN, now named IFN-delta. It was previously shown that the uterus is the most probable target of the pig trophoblastic IFNs, since no autocrine effect was found on the trophoblast. It has also been shown that, unlike for the ruminant species, the pig trophoblastic IFNs do not play an apparent role in the so-called maternal recognition of pregnancy. We have focused this review on IFN-gamma, because first, it is the major species secreted and secondly, IFN-gamma has various regulatory effects on different tissues, including lymphoid cells. We particularly address the question of the possible role of trophoblastic IFN-gamma in early pregnancy, in the light of the known biological functions of human and mouse IFN-gamma. PMID- 11944805 TI - Use of a serum-free medium to produce in vitro Neospora caninum and Toxoplasma gondii tachyzoites on Vero cells. AB - Neospora caninum and Toxoplasma gondii are cyst-forming coccidian parasites of human and veterinary clinical relevance. In vitro cultivation of the protozoans using Vero cells is usually performed in order to produce antigenic materials. Quantitative and qualitative comparisons of Vero cells grown in RPMI medium supplemented either with foetal calf serum (FCS), horse serum (HS) or a specific serum-free additive (DefCell) were performed. A serum-free cell culture system used to propagate N. caninum (NC-1 isolate) and T. gondii tachyzoites (Rh stain) were compared with the other two cell culture systems. FCS supplemented media was found to be more effective than the others in promoting Vero cells and N. caninum tachyzoites. However, it was found unable to support adequate T. gondii tachyzoite proliferation. Vero cells, T. gondii and N. caninum tachyzoite production gave similar growth patterns with either HS or DefCell supplemented media. Defcell was considered as a good alternative to supplement culture medium. PMID- 11944806 TI - Occurrence of respiratory disease outbreaks in fattening pigs: relation with the features of a densely and a sparsely populated pig area in France. AB - A survey was carried out in France in 1999 in a Densely Populated Pig Area (DPPA) and a Sparsely Populated Pig Area (SPPA) from which 80 and 55 pig farms were respectively investigated. The two areas were compared regarding the number of respiratory disease outbreaks in fattening pigs on each farm per year with a multiple correspondence analysis and hierarchical clustering. The two areas exhibited different typologies: in the DPPA, high density was associated with a high proportion of finishing-pig units resulting in much moving of piglets within the area. Farrow-to-finish farms located in the DPPA had many contacts with external vehicles; this was associated with more than two respiratory disease outbreaks per farm per year. There also was a lack of biosecurity measures implemented on these farms. Conversely, the good health situation of the farms located in the SPPA was associated with few external contacts and good biosecurity measures. In a second step, risk factors for the occurrence of more than 2 respiratory outbreaks per year were studied. Having more than 4 pig farms within a 2 km radius area, more than 30 incoming rendering trucks per year, and storage of the carcasses of dead animals within the farm yard perimeter increased the risk of occurrence of more than 2 respiratory disease outbreaks per year on the farm. This risk was also increased when there were more than 2 animal transport lorries entering the farm per month and more than 1 veterinarian's or technician's vehicle coming in every 2 months. These results were discussed because of possible bias due to the retrospective design of the survey and the sampling scheme (randomisation within two areas). PMID- 11944807 TI - Activation of phagocytes during initiation and resolution of mammary gland injury induced by lipopolysaccharide in heifers. AB - The object of the study was the comparative assessment of phagocyte activation during initiation and resolution of mammary gland injury induced by lipopolysaccharide (LPS) or buffered salt solution (PBS) on the basis of the CD14 receptor positivity. The experiments were carried out in 15 clinically normal Holstein x Bohemian Red Pied crossbred heifers, aged 14 to 18 months. Noninflammatory and inflammatory mammary gland injury were induced by intramammary administration of PBS (10 mL) and LPS (10 mL, 1 microg/mL), respectively. Samples of the cell populations were obtained by mammary lavages at 24 h intervals. Flow cytometry was used to determine the CD14+ neutrophils, monocytes, and macrophages. The percentage of CD14+ neutrophils was only 1.2% and 1.3% 24 h after the treatment with PBS and LPS, respectively. The resolution was accompanied by an increase in proportion of CD14+ neutrophils. The proportion of CD14+ neutrophils returned to initial values in the PBS-treated, but not in the LPS-treated mammary glands till 96 h. Percentage of CD14+ monocytes increased after 24 h and the effect was more pronounced in the LPS-treated than in the PBS treated mammary glands (P < 0.05). The percentage of CD14+ macrophages decreased highly significantly at 24 h in the LPS-treated, but not in the PBS-treated mammary glands (P < 0.01). The resolution of mammary gland injury (48 to 96 h) was characterised by an increase in CD14+ macrophages proportion, which was greater in the LPS-treated than PBS-treated mammary glands (P < 0.01). The activation of macrophages during resolution of mammary gland injury can be interpreted as an important mechanism of restitution. PMID- 11944808 TI - Prevalence of Bartonella infection in domestic cats in Denmark. AB - Whole blood and serum from 93 cats (44 pets and 49 shelter/stray cats) from Denmark were tested for the presence of feline Bartonella species by culture and for the presence of Bartonella antibodies by serology. Bartonella henselae was isolated from 21 (22.6%) cats. Bacteremia prevalence was not statistically different between shelter/stray cats (13/49, 26.5%) and pet cats (8/44, 18.2%), but varied widely by geographical origin of the cats, even after stratification for cat origin or age (p < 0.001). All isolates but one were B. henselae type II. The only cat bacteremic with B. henselae type I was not co-infected with B. henselae type II. None of the cats was harboring either B. clarridgeiae or B. koehlerae. Almost half (42/92, 45.6%) of the cats were seropositive for B. henselae and antibody prevalence was similar in shelter/stray cats (23/49, 46.9%) and pet cats (19/43, 44.2%). This is the first report of isolation of B. henselae from domestic cats in Denmark. This study also indicates that domestic cats, including pet cats, constitute a large Bartonella reservoir in Denmark. PMID- 11944809 TI - Isolation and characterisation of local strains of Chlamydophila abortus (Chlamydia psittaci serotype 1) from Tunisia. AB - Chlamydiosis is one of the major diseases that can lead to abortion in ewes. Since 1997, in 5 regions of Tunisia, Chlamydia-related abortions have been reported in 15 sheep and goat flocks. One hundred and sixty-six sera and 50 vaginal swab samples were collected from adult ewes. Chlamydial antigens were detected in 29 (58%) of the vaginal swabs using Enzyme Linked Immunosorbent Assay (ELISA) while 9 (18%) were positive by cell culture. Five strains were recovered from 4 different sheep flocks. Monoclonal antibody profiles and restriction fragment length polymorphism (RFLP) analysis of the 16S-23S rRNA spacer region showed that these isolates were C. abortus. Using amplified fragment length polymorphism (AFLP), these Tunisian strains were shown to exhibit the same pattern as strains isolated in France. PMID- 11944810 TI - Escherichia coli heat-stable enterotoxin b (STb) in vivo internalization within rat intestinal epithelial cells. AB - Heat-stable enterotoxin b (STb) is a low molecular weight toxin known to bind sulfatide, its receptor. The fate of STb bound to rat intestinal epithelium cells was followed using an anti-toxin gold labeled assay and transmission electron microscopy. The data suggest that STb toxin and the fusion protein maltose binding protein (MBP)-STb were internalized whereas its mutant I41 E-M42R with reduced hydrophobicity did not show internalization. There was a significant difference in the mean of gold particles per field between rat intestine incubated with STb or the fusion protein MBP-STb and the negative control consisting of intestine incubated with PBS alone. No subcellular compartment seems to be particularly aimed by the toxin as gold particles were randomly distributed within the cell. PMID- 11944811 TI - The health care professional as a manager: finding the critical balance in a dual role. AB - The health care professional who assumes a management role must recognize that he or she is adopting a second and concurrent career of equal importance to his or her primary occupation. Many such managers have considerable difficulty balancing the two sides of the role because most are well trained in their specialties but enter management with little or no preparation for management. Lack of preparation and inadequate understanding of the requirements of the management side of the role lead to discomfort in management matters and in turn frequently cause a manager to seek refuge in being more of a specialist than manager. The most successful managers will be those who develop the ability to appropriately balance the sides of the dual role. PMID- 11944812 TI - The growing acceptance of complementary and alternative medicine. AB - Alternative and complementary medicine is becoming more popular among consumers and prescribed more by health care professionals. Alternative medicine can be traced back thousands of years, however, it wasn't introduced to the United States until the early 1900s. Alternative medicine encompasses a wide range of therapies including homeopathy, naturopathy, chiropractic, massage and bodywork therapy, meditation, nutritional supplements, and herbal remedies. Understanding the background and benefits of these alternatives is important to all health care professionals. PMID- 11944814 TI - Taking charge by "letting go". AB - An administrator who is able to delegate to subordinates without feeling the need to "micromanage" gains power by "letting go." Power is the ability to influence others by persuasion in coercion. The administrator who is able to "let go" can enjoy the role of facilitator and mentor to subordinates. Details that were formerly obsessed about can be left to others. The administrator who has accomplished the art of "letting go" can say of a subordinate manager, "I trust the manager as a competent professional. I don't 'supervise' constantly. That would be 'micromanaging.' If that person needs some assistance, I will encourage the request." PMID- 11944813 TI - Health care reform and the pharmaceutical industry: crucial decisions are expected. AB - For the past 30 years, the largest growing segment of the United States economy is the health care industry. The United States is in a transitional period as American citizens born between 1946 and 1964, the Baby Boomer generation, reach retirement age. In recent years, pharmaceutical costs have been rising faster than the inflation rate, leaving the American public to ask many questions. A major area of interest to policymakers regarding the health care reform agenda is patient spending on pharmaceutical items. Government-funded programs such as Medicare and Medicaid are facing the possibility of running out of funds and require substantive reform. Pharmaceuticals are not covered under the basic Medicare programs. As a result, senior citizens are forced to cover their prescription expenses out of pocket or purchase supplemental insurance plans. This extra expense is leaving many senior citizens across the country struggling to support their ongoing medical needs. PMID- 11944815 TI - Roles of managers in academic health centers: strategies for the managed care environment. AB - This article addresses survival strategies of academic health centers (AHCs) in responding to market pressures and government reforms. Using six case studies of AHCs, the study links strategic changes in structure and management to managerial role performance. Utilizing Mintzberg's classification of work roles, the roles of liaison, monitor, entrepreneur, and resource allocator were found to be used by top-level managers as they implement strategies to enhance the viability of their AHCs. Based on these new roles, the study recommends improving management practices through education and training as well as changing organizational culture to support management decision making and foster the continued growth of managers and their AHCs. PMID- 11944816 TI - A macro perspective of non-clinical student internship programs. AB - Internships are advantageous to both health care organizations and students. Intern employers benefit in many ways such as completing meaningful backlogged projects, savings on benefits, using internships as a recruiting tool, and becoming a partner in the educational process. There are drawbacks to internships such as increased managerial time to supervise interns and monitor projects, contingent workforce issues with which to deal, and assignment of routine tasks only. Indicators for administration of meaningful internship programs as well as evaluation concerns are discussed. This article addresses college and university internships excluding applied health care clinical rotations. PMID- 11944817 TI - New paradigms in drug design and discovery. AB - The new millennium has ushered in an era of science that will revolutionize a great majority of our daily activities. That revolution is being experienced by a growing number of the population who are pushing the average life expectancy closer to the 80-year mark. The primary reason for this increase is the changes we have made in the last 2-3 decades both in how we live our lives as well as how we treat our maladies when they arise. The advent of new techniques in diagnostics and surgery have allowed many to survive debilitating illnesses when their chances would have been slim only a few years ago. In addition, several new therapeutic agents have been developed in the latter part of the 20th century that have improved our quality of life and increased our overall survival time. New medicines to treat cardiovascular, degenerative, infectious, and neoplastic diseases are rapidly being discovered in an effort to further lengthen our lifetimes. The processes used by academic and industrial scientist to discover new drugs has recently experienced a true renaissance with many new and exciting techniques being developed in only the past 5-10 years. In this review, we will attempt to outline these latest protocols that chemists and biomedical scientist are currently employing to rapidly bring new drugs to the clinic. PMID- 11944818 TI - Application of high-throughput, molecular-targeted screening to anticancer drug discovery. AB - Increasing insight into the genetics and molecular biology of cancer has resulted in the identification of an increasing number of potential molecular targets for anti-cancer drug discovery and development. These targets can be approached through exploitation of emerging structural biology, "rational" drug design, screening of chemical libraries, or a combination of these methods. In this article we discuss the application of high-throughput screening to anti-cancer drug discovery, with special reference to approaches used at the U.S. National Cancer Institute. PMID- 11944819 TI - Impact of recombinant DNA technology and protein engineering on structure-based drug design: case studies of HIV-1 and HCMV proteases. AB - Structure-based drug design is an organized, multidisciplinary endeavor undertaken by scientists from many different scientific fields. The success of structure-based drug design was only made possible by advances in structure biology that provides the three-dimensional structure of the drug design target with which small molecular chemical ligands interact. Visualization of the conformation and interactions of a small molecule ligand bound to the protein target in the co-crystal structure of the protein:ligand complex enables the design of new chemical compounds with improved binding affinity and specificity. With the advances in molecular biology, lab automation, and computational science, genomic data have now become available for the human genome, as well as various other organisms. The pharmaceutical industry is currently putting forth tremendous effort in the area of functional genomics and structural genomics in attempts to decipher functions and structures of protein encoded by genes, with the ultimate goal of identifying novel targets for drug discovery and development. This chapter discusses the significant impact made by recombinant DNA technology and protein engineering on structural biology and, more specifically, on structure-based drug design. PMID- 11944820 TI - Photoaffinity labeling in drug discovery and developments: chemical gateway for entering proteomic frontier. AB - One of the major events occurring at biological interfaces is the specific recognition of bioactive ligands by their receptor proteins. The elucidation of interacting partners is an immediate entrance into the discovery of medicinal leads. The method of photoaffinity labeling enables the direct probing of target protein through a covalent bond introduced between a ligand and its specific receptor. Thus, the photoaffinity labeling is applied in two stages of drug discovery and development processes. First, the method is useful for the screening of early leads. If the binding site analysis of target protein is important for defining a particular pharmacophore, the photoaffinity labeling will give the structural information at the contact point of drugs with receptors. Second, emerging new technologies, combinatorial chemistry, recombinant DNA techniques, and high-throughput analysis, are extending the potential of photoaffinity labeling to become a rapid and more sensitive means for the identification of drug-receptor pairs as well as the elucidation of molecular recognition mechanism at drug-receptor interfaces. This review focuses on several recent impacts of photoaffinity labeling as a useful tool for drug discovery and developments. PMID- 11944821 TI - RNA and RNA-protein complexes as targets for therapeutic intervention. AB - Today, the majority of pharmaceuticals developed to treat cancers and viral/bacterial infections target cellular, bacterial or viral proteins known to be associated with a given pathology. Although proteins are the focus of most current drug discovery efforts, exciting new research has recently begun which aims to exploit ribonucleic acid (RNA) and RNP particles as novel targets for pharmaceutical development. These RNA-targeted research efforts have been fueled by an increased appreciation for the central role played by RNA and RNA-protein interactions in many biological processes and diseases, together with a better understanding of RNA structure and an improvement in biophysical/biochemical techniques available to study RNA. As for protein targets, genome sequencing is greatly accelerating the identification of human and microbial RNA transcripts for targeted drug discovery. With this explosion in the number of potential RNA and RNP targets, the effective development of specific small molecule RNA-based drugs requires robust and general approaches for detecting and quantifying RNA ligand interactions, which can be used as high-throughput screens (HTS) and for obtaining rapid structural information to guide rational drug design. In this review, an overview of the potential for therapeutic intervention based on RNA and RNP targets is presented, together with recent efforts to develop generally useful nuclear magnetic resonance (NMR) and fluorescence binding assays for screening and optimizing drugs aimed at RNA and RNP targets. PMID- 11944822 TI - Inhibitors of protein farnesyltransferase as novel anticancer agents. AB - This paper describes recent progress in the design, synthesis and biological evaluation of inhibitors for the enzyme protein farnesyltransferase (PFTase). This enzyme plays a critical role in the post-translational modification of a range of different intracellular proteins. In particular, PFTase attaches a farnesyl group to the GTPase Ras whose oncogenically mutated form is found in over 30% of human cancers. As a result PFTase inhibitors have been developed as potential cancer therapeutic drugs either by rational design based on the structure of the CAAX carboxyl terminus of Ras or random screening of chemical libraries or natural products. Some of these inhibitors show remarkable inhibition potency against PFTase at subnanomolar concentrations and >1000-fold selectivity compared to the related enzyme geranylgeranyltransferase-I. Certain of these compounds are highly effective at blocking the growth of human tumors in animal models and are now undergoing clinical trials. However, several issues in the research remain unsolved, including the mechanism by which PFTase inhibitors suppress tumor growth. Although it has been established that PFTase inhibitors block prenylation of Ras in vitro, the results in wholecells and animal studies suggest the possibility that proteins other than Ras are affected. PMID- 11944823 TI - Cyclization strategies in peptide derived drug design. AB - The choice of peptides as prototype modulators of biological function, is justified on the grounds that peptides are natural constituents of living systems. They exist as hormones, biochemical inhibitors, antigens, growth factors, transmembrane carriers and, indeed they are comprised of the building blocks of all proteins. As such, the natural and mutated analogs of these functional entities provide a rich variety of pharmacophore models for further development. Peptidomimetic modification of active peptides can provide biostable analogs. Moreover, cyclization of linear peptides is frequently used as an attractive venue to provide both conformationally more restricted as well as more biostable analogs. The objective of this review is to report an updated summary of the more recently developed methodologies for the design and synthesis of cyclized peptides, citing selected examples of the effect of cyclization on both proteolytic stability and biological activity. PMID- 11944824 TI - Identification of epoxyhenicosadiene and novel diepoxy derivatives as sex pheromone components of the clear-winged tussock moth Perina nuda. AB - Four EAG-active components (A-D) were found in the solvent extract of virgin females of the clear-winged tussock moth, Perina nuda. The most abundant component (B, ca. 250 ng/female) was identified as (3Z,6S,7R,9Z)-6,7 epoxyhenicosa-3,9-diene by GC-MS analyses of the extract, chemical derivatization, and comparative chiral HPLC. Minor components also elucidated were (3Z,9Z)-cis-6,7-epoxyicosa-3,9-diene, (A); (3R,45,6S,7R,9Z)-3,4-6,7 diepoxyhenicos-9-ene, (C); and its 3S,4R,6S,7R isomer, (D); with amounts of 0.4, 5, and 8 ng/female, respectively. Component B showed weak attractiveness to male moths in the field. The attractiveness was significantly enhanced by addition of component(s) C and/or D. No males were captured with either the antipode of component B or its mixtures with the minor components. In this field test, noctuid Hypocala rostrata males were also attracted with the synthetic P. nuda pheromone. PMID- 11944825 TI - 2-Methyl-(Z)-7-octadecene: sex pheromone of allopatric Lymantria lucescens and L. serva. AB - Our objective was to identify the sex pheromone of Lymantria lucescens and Lymantria serva (Lepidoptera: Lymantriidae), whose larvae defoliate, respectively, Quercus spp. in temperate regions and Ficus spp. in the subtropics. Coupled gas chromatographic-electroantennographic (GC-EAD) detection analyses of pheromone gland extracts revealed one EAD active compound produced by female L. lucescens and by female L. serva. This was identified as 2-methyl-(Z)-7 octadecene (2me-Z7-18Hy) by retention index calculations on DB-5, DB-23, and DB 210 columns and by comparative GC-mass spectrometric (MS) and GC-EAD analyses of the insect-produced candidate pheromone and synthetic 2me-Z7-18Hy. In field experiments, traps baited with 2me-Z7-18Hy captured male L. lucescens near Toyota City, Japan, and male L. serva in Taipei, Taiwan. Allopatric distribution of L. lucescens and L. serva seems to allow both species to use the same sex pheromone without compromising its specificity. PMID- 11944827 TI - Semiochemicals of the Scarabaeinae: VI. Identification of EAD-active constituents of abdominal secretion of male dung beetle, Kheper nigroaeneus. AB - Using gas chromatography with flame ionization detection (FID) and electroantennographic detection (EAD) in parallel, and employing chiral and achiral capillary columns, three constituents of the abdominal sex-attracting secretion of male Kheper nigroaeneus dung beetles were found to elicit reproducible EAD responses in male and female K. nigroaeneus antennae. One of these constituents is present in the secretion in such a small quantity that it could not be detected by FID, and it was not identified. The other constituents were identified as 3-methylindole (skatole) and (R)-(+)-3-methylheptanoic acid. PMID- 11944826 TI - Activity of male pheromone of Melanesian rhinoceros beetle Scapanes australis. AB - Laboratory and field investigations were carried out to investigate the nature and role of the male pheromone emitted by the Dynast beetle Scapanes australis and to develop a mass trapping technique against this major coconut pest in Papua New Guinea. We report the biological data obtained from natural and synthetic pheromone, previously described as an 84:12:4 (w/w) mixture of 2-butanol (1), 3 hydoxy-2-butanone (2), and 2,3-butanediol (3). EAG recordings from natural and synthetic pheromone and a pitfall olfactometer were poorly informative. In contrast, extensive field trapping trials with various synthetic pheromone mixtures and doses showed that 1 and 2 (formulated in polyethylene sachets in 90:5 v/v ratio) were necessary and sufficient for optimum long-range attraction. Beetles were captured in traps baited with racemic 1 plus 2, with or without a stereoisomer mixture of 3 (2.5- to 2500-mg/day doses). Plant pieces, either sugarcane or coconut, enhanced captures by the synthetic pheromone, which was active alone. Traps with the pheromone caught both sexes in a 3:2 female-male ratio. A pheromone-based mass trapping led to the capture of 2173 beetles in 14 traps surrounding 40 ha of a cocoa-coconut plantation. The captures followed a log-linear decrease during the 125-week trapping program. The role of the male pheromone and its potential for crop protection are discussed. PMID- 11944828 TI - Identification of the sex pheromone of Holotrichia reynaudi. AB - The male attractant pheromone of the scarab beetle Holotrichia reynaudi, an agricultural pest native to southern India, was extracted from abdominal glands of females with hexane and analyzed by gas chromatography-mass spectrometry. Field testing of the candidate chemicals, indole, phenol, and anisole, both alone and as binary mixtures, led us to conclude that anisole was the major component of the sex pheromone. Neither male nor female beetles were attracted to indole or phenol on their own. Similarly, when indole and anisole were combined, the attractiveness of the solution did not increase over that obtained with anisole alone. However, combination of phenol and anisole did alter the attractiveness of anisole, with fewer male beetles attracted to the binary mixture than to anisole on its own. The behavior of female beetles was not altered by any of the chemicals tested. Anisole is also the sex pheromone of H. consanguinea, making this the first known example of two melolonthine scarabs sharing the same pheromone. PMID- 11944829 TI - Is dimethyldecanal a common aggregation pheromone of Tribolium flour beetles? AB - Flour beetles are cosmopolitan and common pests in grain stores and flour mills. Their ability to exploit a wide variety of stored products has contributed to their status as major pests of stored food. Although it was previously reported that the same aggregation pheromone, 4,8-dimethyldecanal (DMD), is shared by three flour beetles species (Tribolium castaneum, T. confusum, and T. freemani), the volatiles released by the other Tribolium species associated with stored products have not yet been examined. In the present study, the volatiles produced by males and females of eight Tribolium species were examined by solid phase microextraction (SPME). SPME samples were analyzed by gas chromatography coupled to mass spectrometry (GC-MS). Experiments were conducted to identify volatiles emitted by the adults of different Tribolium species and to determine whether DMD is a common aggregation pheromone. We observed that DMD is not a common pheromone of the eight species tested, but is common to T. castaneum, T. confusum, T. freemani, and T. madens. Two other volatiles were detected, 1-pentadecene, which is shown here to be a common semiochemical of flour beetles, and 1,6 pentadecadiene, which was detected in five species (T. audax, T. brevicornis, T. destructor, T. freemani, and T. madens). PMID- 11944830 TI - Female sex pheromone polymorphism in adzuki bean borer, Ostrinia scapulalis, is similar to that in European corn borer, O. nubilalis. AB - Individual analysis of the female sex pheromone of the adzuki bean borer, Ostrinia scapulalis, has shown that the sex pheromone of this species comprised (E)-11-tetradecenyl acetate (E11-14:OAc) and (Z)-11-tetradecenyl acetate (Z11 14:OAc) at variable blend ratios. The pheromone blend could be tentatively categorized into three types with respect to the proportion of E11-14:OAc: E type (94-100%, median 99.2%), Z type (0-16%, median 3.0%), and intermediate type (I type, 48-85%, median 63.7%). In addition to the identity of components, the blend ratios in the three types were similar to those of the E strain, Z strain, and hybrid of the European corn borer, O. nubilalis, respectively. This finding suggests that two closely related but morphologically distinct species, O. scapulalis and O. nubilalis, share almost the same sex pheromone communication systems. The significance of this similarity in the two sibling species is discussed. PMID- 11944832 TI - Brook char (Salvelinus fontinalis) can differentiate chemical alarm cues produced by different age/size classes of conspecifics. AB - A wide diversity of aquatic organisms release chemical alarm cues when captured by a predator. For most animals, it is not known whether the specific chemicals that comprise the alarm cue are conserved as prey animals age. In this study, we tested whether brook char (Salvelinus fontinalis) can differentiate alarm cues produced by individuals of different ages/sizes. In separate laboratory experiments we exposed small brook char and large brook char to chemical alarm cues from small brook char, large brook char, and a control of swordtails (Xiphophorus helleri). Both small and large brook char responded with antipredator behavior to chemical alarm cues from both small and large char, but not to those from swordtails. Small char responded with a greater response intensity to cues of small char than to cues of large char. In contrast, large char responded with a greater response intensity to cues of large char than to cues of small char. These results suggest that chemical(s) that act as the alarm cue for fish of different age/size classes may be: (1) identical and that there may be other chemical(s) that allow the test fish to distinguish between cues from fish of different ages/sizes, or (2) the cues are not identical, but similar enough to be recognized. PMID- 11944831 TI - Experimental evidence of an age-specific shift in chemical detection of predators in a lizard. AB - The risk posed by predation is one of the most fundamental aspects of an animal's environment. Avoidance of predators implies an ability to obtain reliable information about the risk of predation, and for many species, chemosensory cues are likely to be an important source of such information. Chemosensory cues reliably reveal the presence of predators or their presence in the recent past. We used retreat site selection experiments to test whether the Australian scincid lizard Eulamprus heatwolei uses chemical cues for predator detection and avoidance. Both adult and juvenile lizards were given the choice of retreat sites treated with scents from invertebrate predators, as well as sympatric and allopatric snake predators. Some of the snake predators were known to eat E. heatwolei, while others did not pose a predation threat. All invertebrate predators posed a risk to juveniles, but not adults because of their size. We found that juvenile E. heatwolei avoided predator odors more strongly than adults. Juveniles avoided both invertebrate predators and snakes, and the strongest response was toward the funnel web spider, the only ambush predator used in this experiment. This result may demonstrate the importance of predator ecology in the evolution of predator detection mechanisms, with chemical cues being more useful in detecting sedentary predators than active predators. Adult lizards showed no avoidance behavior toward predator odors. This result suggests an age specific shift in predator avoidance behavior as lizards get older and become too large for many predators. However, adults showed no response to the odor from the red-bellied black snake, a known predator of adult E. heatwolei. This finding further demonstrates the importance of predator ecology when examining communication between predators and prey. Chemical cues, which are persistent long after predators have vacated the area, may not be useful in detecting the red-bellied black snake, a wide-ranging active forager. PMID- 11944833 TI - Volatile production by buds and corollas of two sympatric, confamilial plants, Ipomopsis aggregata and Polemonium foliosissimum. AB - We used solid-phase microextraction of headspace samples followed by gas chromatography and mass spectrometry to identify and quantify the major volatile compounds produced by the buds and corollas of Ipomopsis aggregata (Polemoniaceae), and we compared them to a sympatric, confamilial plant species, Polemonium foliosissimum. The two species have distinct floral morphologies and pollinators, but share a common predispersal seed predator that oviposits under the calyces of buds. Ipomopsis aggregata buds emitted fewer compounds than the corollas. The buds of I. aggregata were dominated by alpha-pinene and, to a lesser extent, beta-pinene. The corollas of I. aggregata emitted a mixture of 10 compounds. Like the buds, I. aggregata corollas produced relatively high concentrations of alpha-pinene as well as caryophyllene. In addition, the emission of four terpenoids, three esters, and one ketone added to the volatile bouquet of the corollas. Polemonium foliosissimum buds also emitted fewer compounds than the corollas. The buds of P. foliosissimum were dominated by beta pinene and, to a lesser extent, alpha-pinene. The corollas of P. foliosissimum were also dominated by beta-pinene along with three terpenoids, three alcohols, one ester, and two aldehydes. Comparing I. aggregata and P. foliosissimum, bud samples from both species produced considerable amounts of alpha- and beta pinene. Ipomopsis aggregata emitted more alpha-pinene than beta-pinene, while P. foliosissimum emitted more beta-pinene. The corollas of the two species, however, differed in their volatile bouquet. The partition of volatiles between I. aggregata and P. foliosissimum buds and corollas and differences in volatile production between I. aggregata and P. foliosissimum are consistent with selection pressures exerted by organisms interacting with these plants. PMID- 11944834 TI - Learning of herbivore-induced and nonspecific plant volatiles by a parasitoid, Cotesia kariyai. AB - Learning of host-induced plant volatiles by Cotesia kariyai females was examined with synthetic chemicals in a wind tunnel. Wasps were preconditioned by exposure to volatiles and feces simultaneously. A blend of four chemicals, geranyl acetate, beta-caryophyllene, (E)-beta-farnesene, and indole, which are known to be specifically released from plants infested by host larvae Mythimna separata (host-induced blend), elicited a response in naive C. kariyai, but did not enhance the response after conditioning. A blend of five chemicals, (E)-2 hexenal, (Z)-3-hexen-1-ol, (Z)-3-hexen-1-yl acetate, beta-myrcene, and linalool, which are known to be released not only from plants infested by the host larvae, but also from artificially damaged plants or undamaged ones (unspecific blend), elicited little response in naive wasps, but significantly enhanced the wasps' response after conditioning. With a blend of the above nine chemicals, wasps could learn the blend at lower concentrations than they did in the nonspecific blend. Hence, both the host-induced and nonspecific volatile compounds appear to be important for C. kariyai females to learn the chemical cues in host location. PMID- 11944835 TI - Identification of a triterpenoid saponin from a crucifer, Barbarea vulgaris, as a feeding deterrent to the diamondback moth, Plutella xylostella. AB - Larvae of the diamondback moth, Plutella xylostella, a crucifer specialist, refuse to feed on a crucifer, Barbarea vulgaris, because of the presence of a feeding deterrent, which is extractable with chloroform. We isolated a feeding deterrent from B. vulgaris leaves, by successive fractionations with silica-gel, ODS, i.e., C18 reversed phase, and Sephadex LH-20 column chromatographies, and ODS-HPLC, guided by a bioassay for feeding deterrent activity. The structure of the compound was determined to be a monodesmosidic triterpenoid saponin, 3-O-[O beta-D-glucopyranosyl-(1-->4)-beta-D-glucopyranosyl]-hederagenin, based on FAB MS, 1H- and 13C-NMR spectra, and hydrolysis experiments. When the compound was applied to cabbage leaf disks at greater than 0.18 microg/mm2, consumption of the disks by third instars was less than 11% of control disks treated with the solvent alone. Furthermore, all first instars died on the disks treated with the same concentrations. Because the concentration of the compound in the fresh leaves of B. vulgaris was comparable to the effective dose in the cabbage leaf disk tested, we conclude that the unacceptability of B. vulgaris to P. xylostella larvae is primarily due to this saponin. PMID- 11944836 TI - Chemical defense in the plant bug Lopidea robiniae (Uhler). AB - Secretions from the metathoracic glands (MTG) of the black locust bug, Lopidea robiniae (Uhler) (Heteroptera: Miridae) contained six major compounds, including (E)-2-hexenal, (E)-2-hexen-1-ol, (E)-2-octenal, (E)-2-octen-1-ol (E)-2-heptenal, and (Z)-3-octen-1-ol. Males and females did not differ significantly in the relative compositions of identified compounds. In feeding trials, six bird species [robin (Turdus migratorious), blue jay (Cyanocitta cristata), brown thrasher (Toxostoma rufum), killdeer (Charadrius vociferus), starling (Sturnus vulgaris), and house wren (Troglodytes aedon)] demonstrated feeding aversions towards L. robiniae implying that black locust bugs are chemically defended. Bugs discharged the liquid contents of their MTG when attacked, thereby producing a strong and distinct odor. Some birds immediately ejected bugs out of their mouth after biting them, suggesting that the MTG secretion was a deterrent. PMID- 11944837 TI - Screening for effects of phytochemical variability on cytoplasmic protein synthesis pattern of crop plants. AB - Crop plants have to cope with phytochemical variability along with other environmental stresses. Allelochemicals affect several cellular processes. We tested the effect of toxic aqueous leachates from Sicyos deppei, Acacia sedillense, Sebastiania adenophora, and Lantana camara on the radicle growth and cytoplasmic protein synthesis patterns of Zea mays (maize), Phaseolus vulgaris (bean), Cucurbita pepo (squash), and Lycopersicon esculentum (tomato). 2D-PAGE and gel scan densitometry analysis were used to detect differences in cytoplasmic root protein pattern expression. High-, medium-, and low-molecular-weight cytoplasmic proteins were affected by the different aqueous leachates. Crop plant responses were diverse, but in general, an increase in protein synthesis was observed in the treated roots. Maize was the least affected, but both the radicle growth and also the protein pattern of tomato were severely inhibited by all allelopathic plants. The changes observed in protein expression may indicate a biochemical alteration at the cellular level of the tested crop plants. PMID- 11944838 TI - Effects of water deficit stress, shade, weed competition, and kaolin particle film on selected foliar free amino acid accumulations in cotton, Gossypium hirsutum (L.). AB - Leaves of cotton plants, Gossypium hirsutum L., stressed by water deficit, reduced daylight, and weed competition, or treated with a kaolin wettable powder formulation were analyzed for levels of 17 free amino acids (FAAs) using reversed phase high-performance liquid chromatography. Water deficit stress resulted in heightened free proline levels (49.9-fold, P < 0.001) that were correlated with diffusive resistance (seconds per centimeter). Five other FAAs increased, and the amounts of total free essential (for insect growth and development) amino acids and total FAAs also increased (P < or = 0.05). Cotton grown in 50% shade accumulated significantly more free arginine than control plants. In a small-plot weed competition assay, four FAAs increased and three FAAs decreased in association with weed competition, but because free proline levels were not altered and free arginine levels increased, other stresses aside from water deficit, possibly including shading by tall weeds, appear to have caused the changes. In a small-plot kaolin particle film assay, five FAAs were lower in cotton foliage sprayed weekly with kaolin. Because free proline was unaffected and free arginine was lower, it is possible that kaolin's reflectivity heightened light reception. The responses of free proline and arginine to the treatments used in these assays demonstrate that types and degrees of some stresses to cotton can be characterized by accumulations of certain FAAs. The study also demonstrates how some FAA levels can indicate degrees of cotton stress resulting from weed competition and from kaolin particle film application. Porometry and leaf water potential measurements assisted in corroborating some findings of the study. PMID- 11944839 TI - Basic principles of laser Doppler flowmetry and application to the ocular circulation. PMID- 11944840 TI - Optical coherence tomography (OCT): principles of operation, technology, indications in vitreoretinal imaging and interpretation of results. PMID- 11944841 TI - Confocal microscopy of the human cornea in vivo. AB - In vivo, scanning-slit, confocal microscopy offers improved resolution and has resulted in new discoveries of corneal pathology at the cellular level. The ability to provide high resolution, real-time images of the full thickness of the living human cornea gives the clinician and the researcher an important new tool. PMID- 11944842 TI - Today's clinical application of scanning laser technologies. PMID- 11944843 TI - Heidelberg retina tomograph measurements before and after non penetrating surgery. PMID- 11944844 TI - Reliability in the use of the Heidelberg retina tomograph. PMID- 11944845 TI - Normal pressure glaucoma. Open angle glaucoma. PMID- 11944846 TI - Scanning laser polarimetry (SLP) for optic nerve head drusen. AB - Optic nerve head drusen (ONHD) are either clinically invisible or clearly protruding from the disc, in the later case leading to the condition of an irregular, indistinct disc margin or a swollen disc on biomicroscopy. They also may cause visual field defects, even with slow progression. Scanning laser polarimetry (SLP) has been proposed as a rapid, objective and reproducible technology for retinal nerve fiber layer (RNFL) assessment and clinical studies have demonstrated that SLP can help to distinguish between normal and glaucomatous eyes, identify glaucoma suspects and correlates well with visual field defects. The purpose of this study was to evaluate the potential applicability of SLP in 20 consecutive patients with optic nerve head drusen (18 bilateral) that were clinically visible (22 eyes) and invisible (16 eyes). RNFL thickness was studied in patients with and without visual field defects. Patients with visual field defects and ONHD were significantly older and had a small, but significant reduction of visual acuity. Some global SLP parameters (average thickness, ellipse average) were significantly different between subjects with normal and abnormal visual fields. The comparison of the groups with visible and invisible drusen showed that there was no difference in demographic or perimetric data. RNFL thickness measurements were also very similar in both groups. Clinical visibility of drusen was not correlated with RNFL thinning as measured with the GDxTM. SLP assessment, however, was well correlated with functional loss. This objective, non-invasive technology may be an additional option for RNFL evaluation in this condition and an especially useful tool for long-term follow up. PMID- 11944847 TI - Evaluation and definition of physiologic macro cups with confocal optic nerve analysis (HRT). PMID- 11944848 TI - Detecting AMD with multiply scattered light tomography. AB - PURPOSE: to use a novel technique, Multiply Scattered Light Tomography (MSLT), to provide a comfortable, rapid, and noninvasive method for detection and management of Age-related Macualar Degeneration. METHODS: two patient groups were studied in clinical settings with MSLT and confocal scanning laser tomography. In Poway, CA, 21 retinal patients underwent tomography, and the 17 patients with suspicion of exudation also had ICG. An Angio-Scan (Laser Diagnostic Technologies, Inc.) was used to provide simultaneous fundus reflectance and ICG imaging. In Methuen, MA, 20 retinal patients underwent tomography with fluorescein angiography for suspicion of exudation. The MSLT was based on the TopSS (Laser Diagnostic Technologies, Inc.), with a Vertical Cavity Surface Emitting Laser array at 850 mm as the illumination source. The central laser produced confocal images. The surrounding lasers produced multiply scattered light images. RESULTS: MSLT emphasized structures beneath the retina such as drusen, choroidal new vessel membranes, and pigment epithelial detachments. Exudation seen on angiography was visualized by MSLT as topographical structures with distinct borders. Superficial structures, e.g., cysts and epiretinal membranes, were visualized in 850 nm images. DISCUSSION: confocal tomography and MSLT provided a rapid, noninvasive method to detect and localize macular degeneration and pathological structures found in eyes of older patients. PMID- 11944850 TI - Arterial narrowing as a predictive factor in glaucoma. AB - PURPOSE: To evaluate whether retinal arteriolar widths change in normal patients and in different glaucoma types. We measured the arteriolar width at the merge in the optic nerve (MD), at the edge of the optic nerve (ED) and at the peripapillary area (1 disc diameter distance) (disc distance D). HRT software 1.11, Interactive Means program was used. One hundred forty three eyes of seventy two patients divided in four subgroups normal, low-tension glaucoma (LTG), primary open angle glaucoma (POAG) and ocular hypertensive (OH). No statistically significant was found in each studied group. The normal patients showed no changes in the retinal arteriolar width compared with all the glaucoma groups. PMID- 11944849 TI - Large optic nerve heads: megalopapilla or megalodiscs. AB - PURPOSE: To evaluate megalopapillas in order to determine whether they constitute a different population, according to their specific features. MATERIAL: Evaluation of the optic nerve head of 405 eyes divided into 172 normal eyes, 168 pre-perimetric glaucomas (phase 3 or 4), 30 primary congenital glaucomas (children with pure congenital glaucomas operated more than once and diagnosed within the first year of age), and 35 megalopapillas. METHOD: The examinations were performed with the HRT (Heidelberg Retina Tomograph), using a wavelength of 680 nm and the new standard reference plane. Each examination results from the mean of 3 image acquisitions with a SD lower than n = 20. Each optic nerve head was studied in 360 degrees (segments), as well as in quadrants and octants separately (predefined segments). The visual fields were assessed with either the Octopus 1-2-3 or the Octopus 101, programs G2 and G2x (three complete phases). The intraocular pressure was measured by means of daily pressure curves including 7 measurements with applanation tonometry at the office and the first one at 6/7 a.m. with the patient still in bed. Gonioscopic examinations were also performed for classification purposes. RESULTS: By comparing all the stereometric parameters with the Total Area (disc area) and the Rim Volume it has been concluded that the megalopapilla group is different from both the normal control group and the glaucoma group. The disc area of the congenital glaucomas was not statistically different from the megalopapillas, but it was in terms of rim volume and other parameters. CONCLUSION: megalopapilla is an entity characterized by a large optic nerve head which may appear abnormal, with an increased cup, but associated with a normal rim volume, normal visual field and normal IOP. Special attention should be given to its differentiation from pseudoglaucomatous diseases. DISCUSSION: The frequency of megalopapillas seems to be quite higher than in congenital optic nerve head anomalies, but is has been virtually ignored by the literature worldwide so far. PMID- 11944851 TI - Papillary drusen and ocular hypertension. AB - PURPOSE: To evaluate if the changes in the peripapillary and papillary retinal nerve fiber layer, in a young girl who presents papillary drusen and ocular hypertension in both eyes. METHODS: We studied this case with retinography, Humphrey Visual Field, HRT, GDx, and diary curve tonometry. RESULTS: After three years of follow up, no changes were observed in the drusen at the peripapillary and papillary retinal nerve fiber layer. CONCLUSION: In a well controlled ocular hypertensive patient there is no evidence of changes at the optic nerve head and RFNL related to the drusen or to the high pressure. All the diagnosis methods were correlationated with the clinical evolution over time. PMID- 11944852 TI - Non-invasive measurement of the concentration of melanin, xanthophyll, and hemoglobin in single fundus layers in vivo by fundus reflectometry. AB - A new model of the reflection of the human ocular fundus on the basis of the adding-doubling method, an approximate solution of the radiative transport equation, is described. This model enables the calculation of the concentration of xanthophyll in the retina, of melanin in the retinal pigment epithelium and in the choroid, and of hemoglobin in the choroid from fundus reflectance spectra. The concentration values found in 12 healthy subjects are in excellent agreement with literature data. In single cases of pathologic fundus alterations possible benefits to the ophthalmologic diagnostics is demonstrated. PMID- 11944853 TI - Scattering properties of the retina and the choroids determined from OCT-A-scans. AB - GOAL: To determine the coefficient and the anisotropy of scattering as well as the refractive indices in the retina and in the choroid noninvasively in vivo. METHODS: The power of coherent reflected light versus fundus depth is recorded in OCT-A-scans. The ratio of refractive indices is derived from the height of the reflection peaks. Provided that the absorption coefficient is known from fundus reflectometry, the scattering coefficient and anisotropy are calculated from the offset and the slope of the signal behind the reflection peaks on the basis of a single backscattering model. RESULTS: We found scattering coefficients of 12/mm (retina) and 27.5/mm (choroid) as well as anisotropy values of 0.97 (retina) and 0.90 (choroid). DISCUSSION: The OCT is usually employed for the measurement of intraocular distances. The new technique described here gives the unique opportunity to determine further interesting parameters of single ocular layers. The values given above are in good agreement with in vitro results. PMID- 11944855 TI - Functional assessment of the native retinal pigment epithelium after the surgical excision of subfoveal choroidal neovascular membranes type II: preliminary results. PMID- 11944856 TI - Measurement of eye length and eye shape by optical low coherence reflectometry. AB - BACKGROUND: The precise and rapid measurement of eye length and eye shape is essential for investigating eye growth regulation and myopia. For this purpose, we developed an optical low coherence reflectometer (OLCR) and present preliminary measurements. METHODS: The OLCR includes a super luminescent diode (wavelength: 845 nm, coherence length: approximately 30 microm) and rotating glass cube to produce longitudinal scans at a velocity of 0.42 m/s and a repetition rate of approximately 13 scans/s. Heterodyne detection of light reflected from the anterior cornea and the posterior retina permits to measure axial eye length and eye shape (off-axis eye length). Each measurement consists of five consecutive scans. Reproducibility and precision were determined in one volunteer by measuring axial eye length five consecutive times, each time repositioning the eye. Eye shapes were determined in right eyes of four volunteers by measuring eye length every 3.3 degrees from 10 degrees nasally to 10 degrees temporally. RESULTS: Axial eye length measured repeatedly in one volunteer did not differ between or within the measurements (one-factor ANOVA). The average standard deviation was 11 microm. Eye shapes (a) varied substantially among subjects and (b) differed considerably from the corresponding shapes of spherical model eyes with identical axial eye lengths. CONCLUSION: The newly developed OLCR permits the precise and rapid measurement of eye length and eye shape. Such measurements, especially in children, may provide important information about mechanisms of eye growth regulation and the development of myopia. PMID- 11944854 TI - Retinal pigment epithelium translocation and central visual function in age related macular degeneration: preliminary results. AB - PURPOSE: To test the feasibility of a new surgical technique, and to assess visual function over the translocated retinal pigment epithelium (RPE) cells in patients operated upon for subfoveal choroidal neovascularization (CNV) secondary to age-related macular degeneration (AMD). MATERIALS AND METHODS: Six patients presenting previously untreated exudative AMD underwent surgical excision of the subfoveal CNV with RPE translocation and were followed from 1 to 10.5 months. The surgery consisted of a standard three port pars plana vitrectomy (TPPPV), excision of the CNV and RPE translocation. Pre and post-operative ocular examination included best-corrected visual acuity measurement, fundus color stereo photography and fundus fluorescein angiography. Optical coherence tomography (OCT) and confocal laser scanning ophthalmoscopy (cLSO) were performed post-operatively. A cross fixation target and a single-point flashing light were projected on different areas of the posterior pole using a cLSO. Photopic 10-2 perimetry, photopic fine matrix mapping, cLSO microperimetry were also performed pre and post-operatively in four patients. OCT cross-sectional scans and cLSO RPE autofluorescence were recorded to detect the presence of viable translocated RPE. Visual acuity, fixation, photopic 10-2 perimetry, photopic fine matrix mapping and cLSO microperimetry were tested for the presence of central visual function. RESULTS: RPE could be effectively translocated at the time of CNV removal from the edge of the RPE defect to a subfoveal location. OCT showed the translocated RPE as an area of increased optical reflectivity with optical shadowing external to it. cLSO showed autofluorescence of the translocated RPE. The cross fixation target was seen when projected on the translocated RPE. During eccentric fixation, the patients could see a flashing point-target projected on the translocated RPE. Photopic 10-2 perimetry, photopic fine matrix mapping and cLSO microperimetry showed presence of central visual function. CONCLUSIONS: The authors propose that translocation of RPE at the time of CNV removal, from the edge of the RPE defect to a subfoveal location, may have a role in the surgical management of AMD. PMID- 11944857 TI - Comparison of optical coherence tomography and fluorescein angiography in assessing macular edema in retinal dystrophies: preliminary results. PMID- 11944858 TI - Functional imaging of the retinal microvasculature by scanning laser Doppler flowmetry. AB - PURPOSE: to image functionally perfused retinal vessels and to assess quantitatively the intercapillary space of the retinal microvasculature. METHOD: The base of functional imaging and the quantitative assessment of the retinal vasculature is the two-dimensional map of the retina encoded by the laser Doppler frequency shift. By Scanning Laser Doppler Flowmetry (HRF. Heidelberg Engineering) the laser Doppler frequency shift of 16.384 retinal sites (256 pixels x 64 lines, spatial resolution 10 mum) of a retinal area of 2.7 x 0.7 mm was gained. The image processing was performed by a recently described algorithm (AFFPIA). Using the data of the laser Doppler frequency shift of every retinal site, a color-coded retinal image was established showing perfused vessels and capillaries. By automatic pattern analysis of this image vessels and capillaries were identified and segmented. Based on this image the distances in [microm] of every retinal site to the next vessel or capillary were calculated ("distance to next capillary"). The functional imaging of the retinal perfusion was demonstrated in (1) normal retina, (2) retinal arterial occlusion, and (3) proliferative retinopathy. Intraobserver reliability of the quantitative assessment of the parameter "distance to next capillary" was estimated by measuring 10 eyes of 10 subjects at 5 different days by one observer. Interobserver reliability of the quantitative assessment was evaluated by analysing 10 perfusion maps by 5 different operators. In 93 eyes of 71 normal subjects (mean age 40.4 mu 15 years) the juxtapapillary retina was quantitatively evaluated. RESULTS: QUALITATIVE EVALUATION: The functional images of the retinal perfusion of eyes with normal retina, with retinal arterial occlusion, and with proliferative retinopathy corresponded well with the fluorescein angiography. Perfused vessels and capillaries became visible in a high local resolution. QUANTITITATIVE ASSESSMENT: The coefficient of reliability of the introobserver and interobserver reproducibility of the parameter 'mean distance to next capillary" was 0.74, and 0.95, respectively. The quantitative assessment of the perfusion showed that the major part of the retinal sites (>700%) had distances to the next capillary lower than 30 microm 46% of the retinal area had distances to the next capillary from 0-20 microm 26% of the retina had distances from 20-30 microm, 12% of the retina had distances from 30-40 microm 7% of the retina had distances from 40-50 microm, 4% of the retina had distances from 50-60 microm, and 4% of the retinal sites showed distances to the next capillary greater than 60 mum. The mean distance to the next capillary or vessel was calculated with 21 +/- 6.5 microm. CONCLUSION: By non-invasive Scanning Laser Doppler Flowmetry in combination with adequate software it is possible to perform a functional imaging of the retinal vasculature and to measure all index for the functional density of retinal capillaries and vessels. PMID- 11944859 TI - Pulsatile ocular blood flow in patients with pseudoexfoliation. AB - PURPOSE: Pseudoexfoliation syndrome is one of the most frequent causes of open angle glaucoma and is statistically significant associated with a high risk of hypertension, angina, myocardial infarction or stroke and retinal vein thrombosis. The aim of this study was to evaluate the pulsatile ocular blood flow (POBF) in pseudoexfoliation syndrome without (PEX) and with glaucoma (PEG). METHODS: Seventeen eyes with PEX, 17 with PEG and 11 normal eyes of age-matched patients were enrolled. A complete ophthalmological examination included measuring the POBF with the Langham Pneumotonometer as well as the nerve fiber layer thickness by scanning laser polarimetry (GDx). RESULTS: The blood flow parameters, pulse volume and POBF, were statistically significant different between normals and patients with PEG (p < 0.003, t-test). A negative correlation between the intraocular pressure and the POBF was found for all eyes tested. Analysis of GDx? parameters showed a negative correlation for the "number" with the POBF and a positive one for ellipse modulation. CONCLUSION: Although pseudoexfoliation is reported to be a systemic disease measurement of the POBF could not detect any difference between normals and PEX, but was statistically significant different in PEG. Assessments of nerve fiber layer thickness as determined by scanning laser polarimetry also showed a correlation with POBF in some parameters tested. PMID- 11944860 TI - Oximetry with a multiple wavelength SLO. AB - Oximetry may be useful for understanding pathologies of the eye. We use a prototype scanning laser ophthalmoscope capable of simultaneous multiple wavelength imaging to record fundus images. The system is run under non-confocal conditions in using slit apertures with a width of up to 600 microm. The laser lines launched into the SLO were the 633 nm line of a HeNe-laser, and the 815 nm line from a tunable (700 to 900 nm) cw Ti:Sapphire-laser. These wavelengths were selected because of their availability and absorption characteristics. The difference in absorption at these two wavelengths is used to assess blood oxygen content. Images were averaged to improve the signal to noise ratio. This simultaneous method of measuring oxygen content may be preferred to other techniques such as sequential SLO imaging at different wavelengths, or spot analysis using a modified fundus camera. The advantages of this technique are that image registration is not required, and a large area of the retina can be assessed concurrently. PMID- 11944861 TI - A new method for the measurement of oxygen saturation at the human ocular fundus. AB - Measurements of the transmission and the reflectance spectra of whole blood exhibit a different dependence of the thickness of the blood layer. Whereas the transmission spectra decrease at all wavelengths, in the reflectance two ranges with different behaviour have to distinguish. In the absorption dominated range between about 500 and 600 nm, there is only a marginal influence of the thickness of the blood layer. In contrast, in the scattering dominated range higher than about 600 nm the reflectance increases if the thickness of the blood layer increases. Proofed by Monte Carlo simulation, the reflected light, which will be measured in ophthalmologic devices, consists both of light which penetrates a vessel once and will be reflected at the background and of light which is internally reflected at the blood column. Only in small vessels (diameter < 50 microm) the transmitted part dominates. With increasing diameter of the vessels the internally reflected light dominates. Especially under the conditions of a confocal laser scanner, the transmitted part of light is always small. As the measured light consists of both parts of light, a new model for the approximation of the measured reflected light is given, containing an expression for the transmitted and for the internally reflected light. The model was applied for the approximation of the reflectance of blood in a cuvette with an coloured background, delivering sufficient exact result. The oxygen saturation was determined in 256 measurements of 30 healthy persons using an ophthalmo imaging spectrometer. The mean arterial oxygen saturation was 91 +/- 5%, the mean venous oxygen saturation was 59 +/- 9% and the arterio-venous difference in the oxygen saturation was 35%, which is in good agreement with the arterio-venous difference known in the brain. First measurements of the oxygen saturation in age-related macular degeneration point to partly different pathological changes of the retinal microcirculation in dry and in wet AMD. PMID- 11944862 TI - Sildenafil increases ocular perfusion. AB - PURPOSE: Our goal was to determine whether visual system responses to sildenafil accompany shifts in ocular perfusion. The human choroid, which supports the metabolic function of the outer retina, is an erectile tissue, analogous in many respects to the corpus cavenosum. METHODS: Right eyes of 12 normal adults were evaluated before and 2 h after 50 mg oral dose of sildenafil. Pulsatile ocular blood flow (POBF), intraocular pressure (IOP), Heidelberg retinal flowmetry (HRT), 4.26 cpd, 7.5 Hz temporally-modulated contrast sensitivity (CS), full threshold C-20 frequency doubling technology (FDT) perimetry, and blood pressure (BP) were measured. RESULTS: POBF (+29.4%; p > 0.016) and CS (+33.6%; p > 0.014%) increased within 110 (+/-7.7) minutes after sildenafil administration. No subject demonstrated decreases in either variable. HRF flow values increase among 7 of the 9 eyes producing stable scans (+8.2%; p > 0.1). FDT values did not change significantly, nor did systemic pulse amplitude or mean IOP. CONCLUSIONS: Since IOP and systemic pulse amplitude both remained stable after sildenafil administration, while POBF values increased to a level nearly one third greater than baseline. It appears sildenafil can induce intrinsic change in the choroidal vasculature, with an apparently positive impact on pericentral contrast sensitivity. This effect may be of clinical utility. PMID- 11944863 TI - Antiglaucomatous drugs effects on optic nerve head flow: design, baseline and preliminary report. AB - PURPOSE: To determine the effect of the most frequent used antiglaucomatous drugs on the optic nerve head flow. To see the response of the autoregulation system in hypertensive, preperimetric and advanced perimetric glaucomas. MATERIALS: In this preliminary report, optic nerve head parameters and retinal perfusion values were measured in 25 healthy controls as well as in 72 glaucomatous optic nerve heads. The glaucomatous sample was divided into three groups: 24 eyes treated with Betaxolol, 24 with Brinzolamide and 24 treated with Brimonidine (each of these groups was conformed by 8 hypertensive glaucomas, 8 preperimetric glaucomas, and 8 perimetric glaucomas). METHODS: Patients were examined with scanning laser Doppler flowmetry (wave length 638 nm), and flow, volume and velocity indices were assessed in each report. Perfusion maps were analyzed with the new SLDF software, version 3.2 (automatic full field perfusion image analizer). Examinations were also performed with the Heidelberg Retina Tomograph (wave length 638 nm), using the new standard reference plane. Visual fields were performed with the Octopus 123 or 101 perimeters, programs Glx and G2, with 3 phases completed. Diurnal pressure curve with applanation tonometry and gonioscopy (for classification) were also performed. RESULTS: In this preliminary report, it was found that eyes in Hypertensive or preperimetric stages of glaucoma, seem to use their regulation systems in order to preserve their optic nerve head circulation, while advanced perimetric glaucomas seem not be able to preserve their optic nerve head flow. And that antiglaucomatous drugs were not able to increase optic nerve head flow neither. DISCUSSION: Optic nerve head blood supplies is regulated by an autoregulation system. This phenomenon may act in normals, hypertensive and preperimetric glaucomas, but it seems not be able to preserve blood flow in advanced glaucomas. Optic nerve head flow returns to normal values with treatment in glaucoma first stages, while it seems that it could not be restored in advanced glaucomas. PMID- 11944864 TI - Optic nerve head behavior in Posner-Schlossman syndrome. AB - PURPOSE: To analyze the effect of glaucomatocyclitic crisis on the optic nerve head, as well as its consequent structural and hemodynamic changes. To evaluate the surgical indication for Posner-Schlossman syndrome. MATERIALS AND METHODS: Four Posner-Schlossman syndrome cases were evaluated for an analysis of the phenomena occurring in the optic disc by means of retinal confocal tomography and Scanning Laser Doppler Flowmetry, and their correlation with visual function as measured with automated computerized perimetry. In all cases the images were obtained in the mediate period after the attack, while in case 1, measurements were performed in the immediate and mediate periods before, during and after the attack. RESULTS: Significant differences were found between optic nerve head parameters and retinal flow measurements in the different phases evaluated. No permanent optic nerve head damage was demonstrated in any of the cases studied, except for case 4 (Posner-Schlossman syndrome associated with traumatic glaucoma). All the variables analyzed returned to normal values after remission of the acute rise in intraocular pressure. CONCLUSIONS: During the attacks of ocular hypertension the optic nerve head experiences significant morphologic changes and hemodynamic variations; nevertheless since they are transient, they fail to cause permanent damage. There was only one case, in which there was optic disc damage and visual field loss. It can be concluded that surgery should be restricted to those cases with severe and disabling symptoms (relative surgical indication) or to cases with progressive optic neuropathy with visual field loss, when the syndrome is associated with glaucoma (absolute surgical indication). PMID- 11944866 TI - Oral fluorescein angiography with scanning laser ophthalmoscope. AB - OBJECTIVE: To study the efficacy of oral fluorescein angiography compared to intravenous (IV) fluorescein angiography in several retinal diseases. PATIENTS AND METHODS: We performed oral fluorescein angiography with a confocal SLO (Heidelberg Retina Angiograph-HRA) in fourteen psychologically challenged individuals and children, all of whom explicitly refused venipuncture. Patient's diagnosis included diabetes, central serous choroidopathy, toxoplasmosis and choroidal neovascularization. A dose of 30 mg/kg body weight of fluorescein was given. RESULTS: We obtained images of adequate quality to allow interpretation in all cases. CONCLUSIONS: Even though the images were not nearly as good as when obtained after IV injection, we conclude that oral fluorescein angiography with SLO is an appropriate alternative in those cases where a venipuncture is contraindicated by medical or psychological reasons. PMID- 11944865 TI - Imaging the microcirculation of untreated and treated human choroidal melanomas. AB - INTRODUCTION: Histologically demonstrable microcirculation patterns (microcirculation pattern) of human choroidal melanomas have prognostic significance for the patient. We report on our experience in imaging these microcirculation pattern in vivo using simultaneous confocal Fluorescein (FA)- and Indocyaninegreen (ICG) angiography before and after brachytherapy. PATIENTS AND METHODS: The simultaneously procured confocal FA- and ICG angiograms of 50 patients with untreated choroidal melanomas were studied for the visibility of microcirculation pattern. Patients were also followed with simultaneous FA/ICG after brachytherapy. RESULTS: Confocal FA disclosed signs of tumor vascularization in 12 (24%) of the 50 examined patients but microcirculation pattern only in 3 patients (6%). In contrast, simultaneously obtained confocal ICG disclosed microcirculation pattern in 47 patients (94%). In 10 (77%) of the 13 patients the tumor microcirculation changed considerably after brachytherapy: Distortion, thickening, thinning, as well as complete obliteration of vessels could be observed. CONCLUSION: Histologically demonstrated microcirculation pattern can be imaged in vivo. This offers the possibility to assess the likely biologic behavior of the individual tumor without the need for obtaining a cytologic or histologic specimen via enucleation or fine-needle biopsy. Confocal ICG angiogiography images microcirculation pattern better than FA which can be explained by the different absorption-, fluorescence- and exudation characteristics ICG. Follow-up with confocal ICG of choroidal melanomas after brachytherapy shows different features and allows for visualization of the microcirculation reaction to the treatment which might be a useful tool for studying the effects of future anti-angiogenesis based tumor therapies. PMID- 11944867 TI - Basic investigations for 2-dimensional time-resolved fluorescence measurements at the fundus. AB - The decay time is characteristic for several natural fluorophores. The determination of the decay time is independent of the fluorescence intensity. As a consequence, a short living weak fluorescence should be detectable also if it is covered by a intensive long-living fluorescence. As the decay time is influenced by the embedding matrix, information about the cellular stage might be possible. The laser scanning technique in combination with the time correlated single photon counting technique seems to be the optimal method for the discrimination of different fluorophores at the fundus according to the decay time. Fields of an equal decay time are presented as a tau mapping. An experimental set up was developed. Until now, only basic experiments were done on structured fluorescent tests, but under the conditions of the living eye. The results are promising. For the separate detection of the most important short life-time of 120 ps of A2E (excitation at 413 nm. emission 450-600 nm) as a putative precursor of lipofuscin in age-related macular degeneration [11], a light source for pulses in the range of about 50 ps is required. PMID- 11944868 TI - Differential imaging in scanning laser ophthalmoscopy. AB - Differential imaging may be useful for understanding pathologies of the choroid. We use a prototype scanning laser ophthalmoscope capable of simultaneous multiple wavelength imaging to record fundus images. The advantages of simultaneous acquisition compared to serial acquisition are, reduced image capture times, and image registration is not required. The system is run under non-confocal conditions in using slit apertures with a width of up to 600 microm. The laser lines launched into the SLO were the 633 nm line of a HeNe- laser, and the 815 nm line from a tunable (740 nm to 840 nm) cw Ti:Sapphire-laser. The difference in absorption at these two wavelengths is used to produce a differential image. We compare conventional imaging with differential spectral imaging for viewing the choroidal vessels. Qualitative results show the contrast of choroidal vessels in differential imaging is improved compared with normal imaging in the region of the optic disk and for the level of pigmentation in the subjects examined. PMID- 11944869 TI - Occult CNV imaging with scanning laser ophthalmoscope. PMID- 11944870 TI - Glaucomatous optic nerve head changes with scanning laser ophthalmoscopy. AB - PURPOSE: To determine whether there are angiographic differences among normal, preperimetric and advanced glaucoma eyes using indocyanine green angiography with SLO. This method was chosen because of its sensibility to detect peripapillary capillary vessels. METHODS: Scanning laser opthalmoscopy was preformed on normal eyes, preperimetric glaucomas and advanced glaucomas. MATERIAL: The authors used a confocal SLO (Heidelberg Retina Angiograph-HRA) CONCLUSION: Several changes may be seen on peripapillary capillary vessels at the different glaucomatous stages. DISCUSSION: In normal subjects HRT shows preservation of the disc/cup area ratio; indocyanine green angiography shows normal prepapillary plexus pattern on the neuroretinal rim and cup. Subjects on glaucomatous preperimetric stage reveal a decrease in the disc/cup area ratio as a result of an increase of the cup area secondary to a reduction of the neuroretinal rim area. ICG at this hipertensive stage shows an increase in prepapillary plexus visualization, which may be a consequence of increased blood flow while autoregulation is still operative. Subjects with advanced glaucoma show prominent decrease in the disc/cup area ratio as well as marked capillary droupout in ICG angiography. PMID- 11944871 TI - Scanning laser ophthalmoscopy for early diagnosis of vitreoretinal interfase syndrome. AB - PURPOSE: To describe the angiographic signs found using scanning laser ophthalmoscopy for the early diagnosis of vitreoretinal interface syndrome. This method is useful to visualize the inner retinal layers, being more sensitive than fundus biomicroscopy. MATERIAL: 61 patients with vitreoretinal interfase syndrome were evaluated. All of them had evidence of this disease using scanning laser ophthalmoscopy but four patients were referred without diagnosis of vitreoretinal interfase syndrome. These patients showed no biomicroscopic signs and diagnosis was made with SLO. METHODS: Confocal scanning infrared laser ophthalmoscope (Heidelberg Retinal Angiograph assembled by Heidelberg Engineering). This SLO uses an infrared diode laser source of 795 nm. CONCLUSION: Patients included were referred with another diagnosis and with this method the correct diagnosis was made. In conclusion scanning laser ophthalmoscopy allows early diagnosis of this pathology for follow-up and treatment. PMID- 11944872 TI - Annotation: Rett syndrome: recent progress and implications for research and clinical practice. AB - BACKGROUND: Rett syndrome was first described 40 years ago as a profoundly disabling condition in girls. METHOD: Over the last 20 years' national surveys, neuropathological and neurophysiological research have steadily improved understanding of its character and natural history. RESULTS: In the last two years identification of the causative mutations in the gene methyl CpG binding protein 2 (Xq28) has led to a sudden expansion in knowledge about the underlying developmental disorder, with important implications for clinical practice and new opportunities to develop more effective intervention. CONCLUSIONS: It is now clear that the disorder occurs in males and females and that there is a wide range in severity. PMID- 11944873 TI - Practitioner review: Diagnosis of autism spectrum disorder in 2- and 3-year-old children. AB - BACKGROUND: Progress has recently been made in the earlier identification of children with autism spectrum disorder (ASD). Whilst being welcome, this progress to earlier referral and diagnosis presents new challenges to clinical practice, including the accuracy and stability of early diagnosis, the utility of standardised assessment instruments with young pre-schoolers and the ability to indicate prognosis. METHOD: A selective review of recent research literature on the characteristic features of ASD in preschool children. RESULTS: Multidisciplinary diagnostic assessment should include detailed information on developmental history, parents' descriptions of the everyday behaviour and activities of the child, direct assessment of the child's social interaction style, including where possible with age peers, and formal assessment of communicative, intellectual and adaptive function. Clinical assessments need to concentrate on the identification of impairments in early non-verbal social communication behaviours that characterise children with ASD from the second year of life, including social orienting, joint attention, imitation, play and reciprocal affective behaviour. The particular pattern of symptoms that presents in a 2-year-old with ASD may differ from that seen at the more prototypic age of 4 or 5 years. In particular, overt repetitive and stereotyped behaviours may be less notable, although where these are seen alongside the social and communicative impairments they are highly indicative of ASD. The use of standardised assessment instruments and the strict application of the DSM and ICD diagnostic criteria need to be employed with caution, as an expert clinical view has been shown to be more accurate. An important aspect of early diagnostic consultation is an open and straightforward approach to the negotiation of the diagnostic view with parents over time. CONCLUSIONS: Earlier diagnosis and rising recognition of ASD have significant implications for primary healthcare and specialist diagnostic and therapeutic services. PMID- 11944874 TI - The Diagnostic Interview for Social and Communication Disorders: background, inter-rater reliability and clinical use. AB - BACKGROUND: The Diagnostic Interview for Social and Communication Disorders (DISCO) is a schedule for the diagnosis of autistic spectrum and related disorders and assessment of individual needs. It enables information to be recorded systematically for a wide range of behaviours and developmental skills and is suitable for use with all ages and levels of ability. In addition to helping the clinician to obtain a profile of each individual's pattern of development and behaviour, the DISCO also enables identification of specific features found in autistic spectrum disorders that are relevant for use with established diagnostic systems. METHOD: This paper describes the historical background of the DISCO, outlines its structure and reports the results of an inter-rater reliability study with parents of 82 children aged 3 to 11 years with autistic spectrum disorder, learning disability, language disorder or typical development. RESULTS: Inter-rater reliability for the items in the interview was high (kappa coefficient or intra-class correlation at .75 or higher). This level of agreement was achieved for over 80% of the interview items. PMID- 11944875 TI - The Diagnostic Interview for Social and Communication Disorders: algorithms for ICD-10 childhood autism and Wing and Gould autistic spectrum disorder. AB - BACKGROUND: The Diagnostic Interview for Social and Communication Disorders (DISCO) is an interviewer-based schedule for use with parents and carers. In addition to its primary clinical purpose of helping the clinician to obtain a developmental history and description of the child or adult concerned, it can also be used to assist in providing a formal diagnostic category. METHOD: In this study we compared two algorithms based on the ninth revision of the schedule (DISCO 9). The algorithm for ICD-10 childhood autism comprised 91 individual, operationally defined items covering the behaviour outlined in the ICD-10 research criteria. The algorithm for the autistic spectrum disorder, as defined by Wing and Gould (1979), was based on 5 DISCO items that represented overarching categories of behaviour crucial for the diagnosis of autistic disorders. The aim of the study was to examine the implications for clinical diagnosis of these two different approaches. Parents of 36 children with clinical diagnoses of autistic disorder, 17 children with learning disability and 14 children with language disorders were interviewed by two interviewers. Algorithm diagnoses were applied to interview items in order to analyse the relationship between clinical and algorithm diagnoses and the inter-rater reliability between interviewers. RESULTS: Clinical diagnosis was significantly related to the diagnostic outputs for both algorithms. Inter-rater reliability was also high for both algorithms. The ICD childhood disorder algorithm produced more discrepant diagnoses than the Wing and Gould autistic spectrum algorithm. Analysis of the ICD-10 algorithm items and combination of items helped to explain the reason for these discrepancies. CONCLUSIONS: The results indicate that the DISCO is a reliable instrument for diagnosis when sources of information are used from the whole interview. It is particularly effective for diagnosing disorders of the broader autistic spectrum. PMID- 11944876 TI - Are jigsaw puzzle skills 'spared' in persons with Prader-Willi syndrome? AB - BACKGROUND: This three-part study examines previous clinical impressions that people with Prader-Willi syndrome have unusual jigsaw puzzle and word search skills. RESULTS: Children with Prader-Willi syndrome showed relative strengths on standardized visual-spatial tasks (Object Assembly, Triangles, VMI) in that their scores were significantly higher than age- and IQ-matched peers with mixed mental retardation, but below those of age-matched normal children with average IQs. In striking contrast, children with Prader-Willi syndrome scored on par with normal peers on word searches, and they far outperformed them on the jigsaw puzzles, placing more than twice as many pieces as the typically-developing group. Within Prader-Willi syndrome, puzzle proficiency was not predicted by age, IQ, gender, degree of obesity, or obsessive-compulsive symptoms, but by genetic subtypes of this disorder. CONCLUSIONS: Findings are discussed in relation to splinter skills in autism, and to cases with autism and chromosome 15 anomalies that include the Prader-Willi region. PMID- 11944877 TI - Working memory in children and adolescents with Down syndrome: evidence from a colour memory experiment. AB - BACKGROUND: This paper reports information on the visual and verbal short-term memory of individuals with Down syndrome. METHODS: Colour memory in 16 children and adolescents with Down syndrome was compared with that of 16 typically developing children matched for receptive vocabulary. It was suggested that focal colours should be remembered more successfully than non-focal colours on the basis that the former could be remembered using a verbal recoding strategy. However, children with Down syndrome, for whom a deficit in verbal short-term memory makes the use of such a strategy unlikely, should remember focal and non focal colours equally well. More importantly, if individuals with Down syndrome have more developed visual memory abilities than control children, they should outperform them in recognising non-focal colours. RESULTS: Although the group with Down syndrome demonstrated significantly better Corsi blocks performance than controls, and displayed similar levels of colour knowledge, no advantage for colour memory was found. Non-focal colours were remembered by individuals with Down syndrome as successfully as focal colours but there was no indication of a visual memory advantage over controls. Focal colours were remembered significantly more successfully than non-focal colours by the typically developing children. CONCLUSION: Their focal colour memory was significantly related to digit span, but only Corsi span was related to focal colour memory in the group with Down syndrome. PMID- 11944879 TI - The structure of oppositionality: response dispositions and situational aspects. AB - BACKGROUND: The Amsterdam Scale of Oppositionality (ASO) is a recently developed self-report instrument to measure the full range of oppositionality. It was used to test the assumption that oppositionality can best be conceptualized as a combination of emotions and behaviors varying across contexts, i.e., with parents, peers and authority figures. METHOD: The sample consisted of 560 boys and 598 girls, aged 8 to 12 years. The thirty items of the ASO, grouped in item parcels, were analyzed using confirmatory factor analyses. RESULTS: Results confirmed the main hypothesis. The best fitting models contained strongly related emotional and behavioral factors and three mutually related situational factors. Oppositionality appeared to be to a large extent situation-specific. Girls are more affected by the situation than boys and show less oppositionality only outside the family context. CONCLUSIONS: Results are discussed with respect to the concept of oppositionality, varying expectations for interpersonal consequences, and implications for clinical assessment and studies of inter informant reliability. PMID- 11944878 TI - The relationship between DSM-IV oppositional defiant disorder and conduct disorder: findings from the Great Smoky Mountains Study. AB - BACKGROUND: We examine models of the relationship between oppositional defiant disorder (ODD) and conduct disorder (CD) in a community sample. Particular attention is paid to the generalisability of findings based on clinic-referred boys. METHODS: The analyses were based on four waves of data from the Great Smoky Mountains Study covering children in the community aged 9-16 years. Child and parent reports of DSM-IV symptoms, diagnoses, and a range of family and environmental adversities were collected using the Child and Adolescent Psychiatric Assessment. RESULTS: Cross-sectional analyses indicated that CD and ODD largely shared similar correlates, although some aspects of parenting appeared more related to CD than ODD. This pattern was broadly similar in boys and girls. Longitudinal analyses confirmed that ODD was a strong risk factor for CD in boys and there was a suggestion that ODD was a stronger risk factor for CD than for other common disorders. Atypical family structure was an important factor in the transition between ODD and CD in boys. In girls ODD provided no increased risk for later CD but was associated with increased risk for continued ODD, depression, and anxiety. CONCLUSIONS: These results are more consistent with a developmental relationship between ODD and CD in boys than girls. PMID- 11944880 TI - Quality of infant-parent attachment as reflected in infant interactive behaviour during instructional tasks. AB - BACKGROUND: The quality of attachment of 127 Dutch 15-month-old infants to their primary caregivers was assessed using an abbreviated version of Ainsworth's Strange Situation. METHODS: In addition, infant-parent interaction was observed during a 12-minute series of instructional tasks both at home and in a lab setting. Infants were rated for 1) negativity, 2) avoidance, 3) compliance, and 4) positive affect shown towards the caregiver. RESULTS: At home, avoidant and disorganised infants showed significantly less compliance and significantly more avoidance and negative behaviour than securely attached children. At the lab, avoidant infants were significantly less compliant and more avoidant than secure infants, while disorganised infants distinguished themselves from secure children by significantly lower compliance and significantly higher negativity scores. The resistant infants did not distinguish themselves from the secure infants on any of the four behavioural scales. CONCLUSION: The results show brief observation of parent-infant interactions to yield valuable information on infant-parent attachment quality. PMID- 11944881 TI - Emotional abuse in early childhood: relationships with progress in subsequent family placement. AB - BACKGROUND: Several studies of family placements have indicated poorer outcomes for singly placed children. Two of our own studies have additionally shown that late placements of children who were actively singled out from siblings and alone in the care system were at increased risk of poor progress in the first year. METHOD: This paper draws on the data available for singly placed children from both studies to explore this phenomenon in more detail. The current analyses are limited by the fact that neither study was designed specifically to explore the effect of singling out or parental rejection, rather it emerged as a characteristic of potential explanatory importance. RESULTS: The findings suggest that a history of being singled out or 'preferentially rejected' by birth parents, particularly in combination with 'false affection' from the child and lower levels of sensitivity in the new parents, was associated with poorer outcomes in the first year. Such children were more likely to show deteriorating behaviour patterns and to have more problems in forming satisfactory relationships with new family members. CONCLUSIONS: Older age at placement was associated with poorer outcome, but only significantly so among those children not classified as false in their displays of affection. PMID- 11944882 TI - Amino acids activated by fengycin synthetase FenE. AB - Fengycin is a lipopeptidic antibiotic produced nonribosomally by Bacillus subtilis F29-3. Synthesis of this antibiotic requires five fengycin synthetases encoded by fenC, fenD, fenE, fenA, and fenB. In this study, we analyze the functions of the enzyme encoded by fenE, which contains two amino acid activation modules, FenE1 and FenE2. ATP-PP(i) exchange assay revealed that FenE1 activates l-Glu and FenE2 activates l-Ala, l-Val, and l-2-aminobutyric acid, indicating that FenE activates the fifth and the sixth amino acids in fengycin. Furthermore, l-Val is a better substrate than l-Ala for FenE2 in vitro, explaining why B. subtilis F29-3 normally produces twice as much of fengycin B than fengycin A, which contains d-Val and d-Ala at the sixth amino acid position, respectively. Results presented herein suggest that fengycin synthetase genes and amino acids in fengycin are colinear. PMID- 11944883 TI - L-delta-(alpha-Aminoadipoyl)-L-cysteine-D-valine synthetase: production of dipeptides containing valine residue at its C-terminus. AB - L-delta-(alpha-Aminoadipoyl)-L-cysteine-D-valine synthetase (ACVS) has been recently studied as a model enzyme for peptide synthetases. It was found that in the absence of alpha-aminoadipic acid but in the presence of several cysteine analogues it was incorporated into several analogue dipeptides upon incubation of the potential cysteine analogues with ACVS. [(14)C]Cysteine was incorporated into the[(14)C]cysteinyl-valine analogue dipeptides. Notably, [(14)C]valine incorporation in the presence of N-acylated cysteine analogues was observed. The alpha-aminoadipic acid activation site is influential, inhibitory or promotive, on the production of these putative dipeptide products. The production of dipeptide analogues, containing valine or analogues at the C-terminus, leads to the speculation that the biosynthetic direction of ACV could be from the C terminus to the N-terminus. PMID- 11944884 TI - beta-Endorphin-like peptide SLTCLVKGFY is a selective agonist of nonopioid beta endorphin receptor. AB - It has been found that beta-endorphin (beta-END) and a synthetic beta-END-like decapeptide Ser-Leu-Thr-Cys-Leu-Val-Lys-Gly-Phe-Tyr (termed immunorphin, IMN) corresponding to the sequence 364-373 of human IgG heavy chain stimulate Con A induced proliferation of T lymphocytes from the blood of healthy donors. [Met(5)]enkephalin ([Met(5)]ENK) and an antagonist of opioid receptors naloxone (NAL) tested in parallel were not active. The stimulating effect of beta-END and IMN on T lymphocyte proliferation was not inhibited by NAL. Studies on receptor binding of (125)I-labeled IMN to T lymphocytes revealed that it binds with high affinity to NAL-insensitive binding sites (K(d) = 7.0 +/- 0.3 nM). Unlabeled beta END completely inhibited the specific binding of (125)I-labeled IMN to NAL insensitive binding sites on T lymphocytes (K(i) = 1.1 +/- 0.2 nM). Thus, beta END and IMN bind to common NAL-insensitive binding sites on T lymphocytes and enhance Con A-induced proliferation of these cells. PMID- 11944885 TI - Oxidative stress in vitiligo: photo-oxidation of pterins produces H(2)O(2) and pterin-6-carboxylic acid. AB - Patients with vitiligo accumulate millimolar levels of H(2)O(2) in their epidermis. The recycling process of (6R)-l-erythro-5,6,7,8-tetrahydrobiopterin in these patients is disrupted due to deactivation of 4a-OH-BH(4) dehydratase by H(2)O(2). The H(2)O(2) oxidation products 6- and 7-biopterin lead to the characteristic fluorescence of the affected skin upon Wood's light examination (UVA 351 nm). Here we report for the first time the presence and accumulation of pterin-6-carboxylic acid (P-6-COOH) in the epidermis of these patients. Exploring potential sources for P-6-COOH revealed that sepiapterin and 6-biopterin are readily photo-oxidised to P-6-COOH by UVA/UVB irradiation. Photolysis of sepiapterin and 6-biopterin produces stoichiometric H(2)O(2) under aerobic conditions, where O(2) is the electron acceptor, thus identifying an additional source for H(2)O(2) generation in vitiligo. A detailed analysis utilising UV/visible spectrophotometry, HPLC, TLC, and mass spectroscopy showed for sepiapterin direct oxidation to P-6-COOH, whereas 6-biopterin formed the intermediate 6-formylpterin (P-6-CHO) which is then further photo-oxidised to P-6 COOH. PMID- 11944886 TI - Kinetics of nitric oxide binding to R-state hemoglobin. AB - Despite earlier work indicating otherwise, some recent reports have suggested that nitric oxide (NO) binds to hemoglobin cooperatively. In particular, it has been suggested that, under physiological conditions, NO binds to the high affinity R-state hemoglobin as much as 100 times faster than to the low-affinity T-state hemoglobin. This rapid NO binding could provide a means of preserving NO bioactivity. However, using a flash-flow photolysis technique, we have determined that the rate of NO binding to normal adult R-state hemoglobin is (2.1 +/- 0.1) x 10(7) (s(-1) M(-1)), which is essentially the same as that reported for T-state NO binding. (c)2002 Elsevier Science (USA). PMID- 11944887 TI - Hyaluronidases and CD44 undergo differential modulation during chondrogenesis. AB - Hyaluronan, a high-molecular-weight glycosaminoglycan of cartilage, is deposited directly into the extracellular space by hyaluronan synthases, while hyaluronan catabolism is mediated by the hyaluronidases. An in vitro cell culture system has been established in which human dermal fibroblasts are induced to undergo chondrogenesis. Here, we describe the differential modulation of the hyaluronidases and the up-regulation of the hyaluronan receptor, CD44, during such chondrogenesis. Dermal fibroblasts, plated in micromass cultures in the presence of lactic acid and staurosporine for 24 h, were then placed in serum free, chemically defined medium. At 3 days, RNA was extracted and RT-PCR performed using primers for the hyaluronidase genes. Marked increase in HYAL1 expression was observed, with only moderate increases occurring in HYAL2 and HYAL3. No expression of HYAL4 and PH-20, the sperm-associated hyaluronidase, was detected. RNA levels correlated well with changes in hyaluronidase enzyme activity. Finally, greater expression and staining for the hyaluronan receptor, CD44s, the standard form, were detected. Differential expression of the somatic hyaluronidases and CD44-mediated hyaluronan turnover play a critical role in cartilage development from mesenchymal precursors. PMID- 11944888 TI - Evidence of both extra- and intracellular cysteine targets of protein modification for activation of RET kinase. AB - By use of a specifically sulfhydryl group-reactive chemical, 1,4-butanediyl bismethanethiosulfonate (BMTS), we studied the localization of oxidative stress responsive target cysteines for activation of a receptor-type protein tyrosine kinase, c-RET. The chemical, which reacted with RET proteins on the cell surface for sulfhydryl-linked aggregation, induced autophosphorylation and activation of RET kinase. When extracellular domain-deleted RET mutant (RET-PTC-1) cells were exposed to BMTS, neither the molecular status nor the activity of the enzyme was affected, suggesting that the target cysteines of BMTS to which cells were exposed for reaction are located in the cysteine-rich region of the extracellular domain of RET kinase. Despite this result, the exposure of a subcellular form of c-RET or RET-PTC-1 kinase isolated by immunoprecipitation to BMTS did induce activation of the enzyme. These results suggest that cysteines in both the extracellular and the intracellular domains of RET can work as target sites of accessible BMTS and possibly other oxidative elements for structural modification and activation of RET kinase. PMID- 11944889 TI - HIV-1 Vif-derived peptide inhibits drug-resistant HIV proteases. AB - Vif, one of the six accessory genes expressed by HIV-1, is essential for the productive infection of natural target cells. Previously we suggested that Vif acts as a regulator of the viral protease (PR): It prevents the autoprocessing of Gag and Gag-Pol precursors until virus assembly, and it may control the PR activity in the preintegration complex at the early stage of infection. It was demonstrated before that Vif, and specifically the 98 amino acid stretch residing at the N'-terminal part of Vif (N'-Vif), inhibits both the autoprocessing of truncated Gag-Pol polyproteins in bacterial cells and the hydrolysis of synthetic peptides by PR in cell-free systems. Linear synthetic peptides derived from N' Vif specifically inhibit and bind HIV-1 PR in vitro, and arrest virus production in tissue culture. Peptide mapping of N'-Vif revealed that Vif88-98 is the most potent PR inhibitor. Here we report that this peptide inhibits both HIV-1 and HIV 2, but not ASLV proteases in vitro. Vif88-98 retains its inhibitory effect against drug-resistant HIV-1 PR variants, isolated from patients undergoing long term treatment with anti-PR drugs. Variants of HIV protease bearing the mutation G48V are resistant to inhibition by this Vif-derived peptide, as shown by in vitro assays. In agreement with the in vitro experiments, Vif88-98 has no effect on the production of infectious particles in cells infected with a G48V mutated virus. PMID- 11944890 TI - Phytochemical inhibition of interleukin-4-activated Stat6 and expression of VCAM 1. AB - Cellular functions induced by cytokine interleukin (IL)-4 and IL-4 signaling through signal transducer and activator of transcription (Stat)6 typify a Th2 type immune response. We investigated the inhibitor effect of the NFkappaB blocker parthenolide in the late-phase, Th2-type immune response. Parthenolide blocked by 90.6 +/- 7.4% the IL-4-induced expression of the endothelial vascular cell adhesion molecule (VCAM)-1, a hallmark of extravasation of very late antigen 4-positive leukocytes. The noncytotoxic concentrations of 10 microM parthenolide left a section of the IL-4 receptor signal transduction intact. Parthenolide did not interfere with the immediate IL-4-induced phosphorylation of endothelial Stat6 on its tyrosine residue Y641 and with tyrosine phosphorylation of the adapter molecule, Jak2-both processes are obligatory for dimerization and nuclear translocation of Stat6. But parthenolide inhibited the Stat6 DNA-binding activity in IL-4-stimulated endothelial cells and inhibited the IL-4-driven activation of a luciferase reporter gene under the control of Stat6-responsive elements (IC(50) 5.11 +/- 0.67 microM). Together, these data suggest an anti-chronic disease profile for the sesquiterpene lactone parthenolide. PMID- 11944891 TI - OC-3, a novel mammalian member of the ONECUT class of transcription factors. AB - Transcription factors of the ONECUT class possess a single cut domain and a divergent homeodomain. They regulate gene networks by controlling the expression of other transcription factors and they play an important role in cell differentiation and metabolism. We identified earlier in mammals HNF-6 (ONECUT 1), the founding member of the class, and ONECUT-2 (OC-2). We have now characterized in the mouse a third ONECUT member, which we call OC-3. Its gene is located on chromosome 10. The sequence of OC-3 (490 residues) displays 51% amino acid identity with HNF-6 and 50% with OC-2. OC-3 has a DNA-binding specificity similar to that of HNF-6 and it is a stimulator of gene transcription. OC-3 mRNA is found in brain, stomach, and upper intestine in the adult and embryonic mouse. Our earlier work on HNF-6 and the expression patterns of the three mammalian ONECUT genes suggest that they all participate to the control of organ development from the foregut and midgut endoderm. PMID- 11944892 TI - Chemical synthesis and kinetic study of the smallest naturally occurring trypsin inhibitor SFTI-1 isolated from sunflower seeds and its analogues. AB - The smallest known naturally occurring trypsin inhibitor SFTI-1 (14 amino acid residues head-to-tail cyclic peptide containing one disulfide bridge) and its two analogues with one cycle each were synthesized by the solid phase method. Their trypsin inhibitory activity was determined as association equilibrium constants (K(a)). Additionally, hydrolysis rates with bovine beta-trypsin were measured. Among all three peptides, the wild SFTI-1 and the analogue with the disulfide bridge only had, within the experimental error, the same activity (the K(a) values 1.1 x 10(10) and 9.9 x 10(9) M(-1), respectively). Both peptides displayed unchanged inhibitory activity up to 6 h. The trypsin inhibitory activity of the analogue with the head-to-tail cycle only was 2.4-fold lower. It was also remarkably faster hydrolyzed (k = 1.1 x 10(-4) mol(peptide) x mol(enzyme)(-1) x s(-1)) upon the incubation with the enzyme than the other two peptides. This indicates that the head-to-tail cyclization is significantly less important than the disulfide bridge for maintaining trypsin inhibitory activity. PMID- 11944893 TI - Different effects of glucose starvation on expression and stability of VEGF mRNA isoforms in murine ovarian cancer cells. AB - Vascular endothelial growth factor (VEGF) has been implicated as a potent regulator of angiogenesis in tumors, and its protein exists as at least five isoforms with distinct biologic activities and clinical significance. Tumors under metabolic stress conditions dramatically increase VEGF expression due to both increased transcription and decreased mRNA degradation. However, it is not known how stress conditions regulate expression of each VEGF isoform. Here, we report a novel Taqman real-time RT-PCR strategy for quantification of all murine VEGF isoforms and find that (1) glucose starvation dramatically up-regulates the mRNA level of all VEGF isoforms, with the three abundant isoforms, VEGF120, VEGF164, and VEGF188, increasing at a similar rate, while the rare isoform VEGF144 is more markedly up-regulated; (2) glucose starvation induces a significant increase of the relative abundance of VEGF144 mRNA, but not the more prevalent isoforms VEGF120, VEGF164, and VEGF188, compared to total VEGF; and (3) the stability of each isoform mRNA differs under the control conditions as well as glucose starvation. The latter significantly stabilizes mRNA of all VEGF isoforms at a different rate, with VEGF144 most significantly stabilized. Our results indicate that under metabolic stress conditions VEGF144 is the most dramatically up-regulated VEGF isoform, probably through mechanism(s) different from the three abundant VEGF isoforms. PMID- 11944894 TI - Macrophage activation includes high intracellular myeloperoxidase activity. AB - Macrophages recovered from the peritoneum of mice, 48 h after concanavalin A administration, are primed and have a higher content of myeloperoxidase (MPO) than resident cells. The increase in MPO content is accompanied by an increased capability of macrophages generate hypochlorous acid and increased peroxidase activity. Contrary to the common sense, neutrophils is not the source of the MPO activity found in primed macrophages since macrophages recovered from mice treated with antigranulocyte antibody preserve the peroxidase activity. Given the broad spectrum of action of MPO, the preservation of MPO in primed macrophages might play a special role in the killing of pathogens and inflammation. PMID- 11944895 TI - An infrared study of the thermal and pH stabilities of the GPI-alkaline phosphatase from bovine intestine. AB - Alkaline phosphatase (EC 3.1.3.1) from bovine intestine mucosa (BIAP) is a homodimeric metalloenzyme, which hydrolyses nonspecifically phosphate monoesters at alkaline pH with release of inorganic phosphate and alcohol. BIAP is either soluble (sBIAP) or membrane-anchored by a glycosylphosphatidylinositol moiety (GPI-BIAP). This anchor might have some contribution in the stabilization of the GPI-linked protein structure. Our purpose was to study the role of the anchor by using two parameters, the enzymatic activity and the protein conformation, which was analyzed by using FTIR spectroscopy. We determined that the two forms of BIAP show some similarities with the previously described structure of alkaline phosphatase isolated from Escherichia coli and human placenta. Meanwhile GPI-BIAP and sBIAP exhibit similar specific activities, the presence of the anchor increases the thermal and pH stabilities of the enzyme activity and conformation. PMID- 11944896 TI - Identification of LPS-binding peptide fragment of MD-2, a toll-receptor accessory protein. AB - Members of the toll-like receptor family are crucial in recognition of microbial pathogens as part of innate immune response. MD-2, an accessory protein to TLR4, present on the extracellular side of the membrane is needed to initiate the signal transduction. We have identified a 15 amino acid region of human MD-2 that contains several features of other lipopolysaccharide (LPS) binding proteins and peptides. In vitro LPS neutralization by this peptide was observed and confirmed by 2D transferred NOESY NMR experiments. NMR experiments have also shown binding of the MD-2 peptide to lipoteichoic acid (LTA) but not to peptidoglycan. Furthermore this peptide inhibited growth of gram-negative and to a lower extent of some gram-positive bacteria. Our results indicate that this region of MD-2 might be responsible for binding of LPS and confirms the role of MD-2 as an accessory protein in LPS signaling bestowing the Toll receptors their specificity. PMID- 11944897 TI - The role of angiostatin in the spontaneous bone and prostate cancers of pet dogs. AB - Angiostatin is a potent inhibitor of angiogenesis generated in cancer-bearing hosts by tumor-derived proteases. Because the naturally occurring bone and prostate cancers of pet dogs provide unique model systems to study factors that regulate cancer progression and tumor dormancy, we investigated the capacity of these tumors to generate angiostatin. We determined that angiostatin fragments are present in urine of dogs with bone cancer. The identity of these fragments was confirmed by comparison of the experimentally determined protein sequence to that of a clone of canine angiostatin. Importantly, these fragments were absent in urine collected from the same dogs after complete surgical removal of the primary tumor. We also demonstrate that canine prostate cancer cells are capable of processing plasminogen to angiostatin in vitro. These findings provide rationale for using spontaneous canine tumor models to isolate endogenous angiogenesis inhibitors and to investigate their therapeutic use against cancer. PMID- 11944898 TI - Role of Cbl in shear-activation of PI 3-kinase and JNK in endothelial cells. AB - Fluid shear stress can activate PI-3 kinase and JNK in vascular endothelial cells. This study was designed to establish the role of Cbl as an upstream molecule in the shear stress activation of PI-3 kinase and JNK. Confluent monolayers of bovine aortic endothelial cells (BAECs) were subjected to a shear stress of 12 dyn/cm(2) over intervals ranging from 0.5 to 30 min. Shear stress increased Cbl phosphorylation to 2.9-fold of control and Cbl association with the regulatory PI-3 kinase subunit p85 to 5.4-fold. The PI-3 kinase activity measured in Cbl-immunoprecipitated complexes increased to 11.7-fold in response to shear, suggesting that the shear stress activation of PI-3 kinase involves its association with Cbl. Furthermore, the shear stress induction of JNK was attenuated by a negative mutant of Cbl. Finally, shear stress caused an activation of PI 3-kinase only in BAECs seeded onto fibronectin, vitronectin, or laminin, but not poly-l-lysine. Our results suggest that Cbl plays a critical role in the shear stress induction of PI 3-kinase and JNK activities, and that this shear-induced activation requires the interaction of endothelial integrins with extracellular matrix proteins. PMID- 11944899 TI - Hormone-sensitive lipase is not required for cholesteryl ester hydrolysis in macrophages. AB - Storage of cholesteryl esters in the cytoplasm of macrophages is one of the earliest and most ubiquitous event observed in the development of arteriosclerosis. Macrophages have an enormous capacity to uptake and store cholesterol in the form of cytosolic cholesteryl ester droplets. These stores are mobilized by the action of a neutral cholesteryl ester hydrolase (NCEH), producing free cholesterol that is either secreted to extracellular acceptors or reesterified. It has been proposed that hormone-sensitive lipase (HSL) is responsible for the NCEH activity in macrophages. The present work shows, however, that peritoneal macrophages from HSL null mice hydrolyze cytosolic stores of cholesteryl esters at a comparable rate to that of peritoneal macrophages from wild-type mice, therefore demonstrating that HSL is not the main NCEH in macrophages. PMID- 11944900 TI - Tributyltin causes cytochrome C release from isolated mitochondria by two discrete mechanisms. AB - Using isolated liver mitochondria we show that low concentrations of TBT (0.5 microM) cause the release of mitochondrial cytochrome c, in the presence of Ca(2+). This is reflected in a rapid loss of membrane potential (DeltaPsi(m)), and a large-amplitude swelling characteristic of mitochondrial permeability transition (MPT). Despite this, the inclusion of cyclosporin A could not prevent the release of cytochrome c. Further, in the absence of Ca(2+), low concentrations of TBT (0.5 microM) resulted in a slow sub-maximal shift of DeltaPsi(m), not characteristic of MPT, which was still paralleled by a release of cytochrome c. Further experiments showed that the loss of DeltaPsi(m) in the absence of Ca(2+) was due to a combination of inhibition of respiration and a direct uncoupling effect on the respiratory chain. Under these conditions, rapid swelling of mitochondria could be demonstrated, due to chloride exchange over the inner mitochondrial membrane. Taken together these data suggest that TBT can induce the release of cytochrome c in intact cells by at least two mechanisms. The first and critical mechanism is initiated immediately the mitochondria sense the presence of TBT and involves a slow loss of DeltaPsi(m) and induction of swelling, which allows release of cytochrome c in a relatively non-specific manner and independently from a rise in [Ca(2+)](i). The second mechanism involves the induction of formal MPT as intracellular [Ca(2+)](i) increases. These data help to explain previous observations in intact lymphocytes demonstrating TBT-induced release of mitochondrial cytochrome c in the absence of a rise in [Ca(2+)](i) (Stridh, H., Gigliotti, D., Orrenius, S., and Cotgreave, I. A. (1999) Biochem. Biophys. Res. Commun. 266, 460-465). PMID- 11944901 TI - The novel Rab11-FIP/Rip/RCP family of proteins displays extensive homo- and hetero-interacting abilities. AB - The Rab11-FIP/Rip/RCP proteins are a recently described novel protein family, whose members interact with Rab GTPases that function in endosomal recycling. To date, five such proteins have been described in humans, all of which interact with Rab11, and one (RCP) also interacts with Rab4. Here, we characterise several of these proteins with respect to their ability to interact with Rab4, as well as their ability to self-interact, and to interact with each other. We now demonstrate that two of the family members-pp75/Rip11 and Rab11-FIP3 do not bind Rab4 and show that several members of the family can self-interact and interact with each other. These interactions primarily involve their C-terminal end which includes the Rab binding domain (RBD) that is contained within a predicted coiled coil, or ERM motif. We identify a new (sixth) member of the protein family, which we propose to name Rab11-FIP4, and report the family evolutionary complexity and chromosomal distribution. Furthermore, we propose that the ability of these proteins to bind each other will be important in effecting membrane trafficking events by forming protein 'platforms,' regulated by Rab11 and/or Rab4 activity. PMID- 11944902 TI - The roles of LAT in platelet signaling induced by collagen, TxA2, or ADP. AB - The work presented here demonstrates that platelets from mice lacking LAT (linker for the activation of T cells) show reversible aggregation in response to concentrations of collagen that cause TxA2/ADP-dependent irreversible aggregation of control platelets. The aggregation defect of the LAT-deficient platelets was shown to be the result of almost no TxA2 production and significantly diminished ADP secretion. In contrast, the LAT deficiency does not affect aggregation induced by high concentrations of collagen because that aggregation is not dependent on TxA2 and/or ADP. Even though ADP and TxA2 provide amplification signals for platelet activation in response to low concentrations of collagen, LAT-deficient platelets hyperaggregate to low levels of U46619, a TxA2 analog, or ADP. Though the mechanism(s) of costimulatory signals by collagen, ADP, and TxA2 remains unidentified, it is clear that LAT plays a positive role in collagen induced, TxA2/ADP-dependent aggregation, and a negative role in TxA2 or ADP induced platelet aggregation. PMID- 11944903 TI - Mechanism of specific recognition of the aidB promoter by sigma(S)-RNA polymerase. AB - Transcription of the Escherichia coli aidB gene is controlled by an Esigma(S) dependent promoter (PaidB) and is poorly transcribed by the Esigma(70) form of RNA polymerase in the absence of additional factors. In this report, we investigate the interaction between PaidB and either the Esigma(70) or the Esigma(S) forms of RNA polymerase in vitro. We show that although Esigma(70) can bind the aidB promoter, its interaction with the promoter results in the formation of an open complex inefficient in transcription initiation and sensitive to heparin challenge. Deletion of the C residue at position -13 of PaidB (Delta-13C) slightly impaired transcription initiation by Esigma(S), consistent with the role of -13C as a specific feature of Esigma(S)-dependent promoters. However, Esigma(S) could still bind and initiate transcription from the Delta-13C mutant aidB promoter more efficiently than Esigma(70), suggesting that sequence elements other than the -13C play an important role in specific promoter recognition by Esigma(S). PMID- 11944904 TI - Pentapeptide amides interfere with the aggregation of beta-amyloid peptide of Alzheimer's disease. AB - Amyloid peptides (Abeta) play a central role in the pathogenesis of Alzheimer's disease (AD). The aggregation of Abeta molecules leads to fibril and plaque formation. Fibrillogenesis is at the same time a marker and an indirect cause of AD. Inhibition of the aggregation of Abeta could be a realistic therapy for the illness. Beta sheet breakers (BSBs) are one type of fibrillogenesis inhibitors. The first BSB peptides were designed by Tjernberg et al. (1996) and Soto et al. (1998). These pentapeptides have proved their efficiency in vitro and in vivo. In the present study, the effects of two pentapeptide amides are reported. These compounds were designed by using the C-terminal sequence of the amyloid peptide as a template. Biological assays were applied to demonstrate efficiency. Modes of action were studied by FT-IR spectroscopy and molecular modeling methods. PMID- 11944905 TI - Differential effects of histone deacetylase inhibitors on interleukin-18 gene expression in myeloid cells. AB - Histone deacetyrase (HDAC) inhibitors induce growth arrest and differentiation of leukemia cell lines and tumor cells derived from a large variety of human tissues. Here we showed that HDAC inhibitors sodium butyrate, TSA, and valproate regulated the expression of Interleukin-18 (IL-18), a cytokine with antitumor and proinflammatory properties, in human acute myeloid leukemia cell lines U937 and HEL. Sodium butyrate increased expression of IL-18 protein and mRNA and activated 1357bp IL-18 gene promoter construct. IL-18 mRNA level was up-regulated by TSA or valproate, which also activated IL-18 full-length promoter. While sodium butyrate or TSA stimulated the 108-bp IL-18 minimal promoter, valproate failed to activate it, indicating that valproate may use a distinct mechanism from sodium butyrate and TSA to activate IL-18 gene expression. PMID- 11944906 TI - Effects of platelet-activating factor, tumor necrosis factor, and interleukin 1alpha on the expression of apolipoprotein M in HepG2 cells. AB - Apolipoprotein M (apoM) is a recently discovered human apolipoprotein predominantly present in high-density lipoprotein (HDL) in plasma, exclusively expressed in liver and in kidney. The function of apoM is yet unknown. The human apoM gene is located in the major histocompatibility complex class III region on chromosome 6. Because many genes located in this region are related to the immune response, we have investigated whether apoM might also be involved in the host inflammatory response. In this study we examined effects of the platelet activating factor (PAF), tumor necrosis factor (TNF-alpha), and interleukin 1alpha (IL-1alpha) on apoM expression in a hepatoblastoma cell line, HepG2 cells. PAF significantly enhanced the apoM mRNA levels and the secretion of apoM in HepG2 cell cultures. The enhancement of apoM secretion is seen at a low concentration of PAF (2 ng/ml), whereas a high concentration of PAF increases both the apoM mRNA levels and apoM secretion. Neither TNF-alpha nor IL-1alpha influenced apoM mRNA level and secretion. Furthermore, Lexipafant, a PAF-receptor (PAF-R) antagonist significantly suppressed the mRNA level and the secretion of apoM in HepG2 cells in a dose-dependent manner. Neither PAF nor Lexipafant influenced the mRNA levels and the secretion of apoA-I, apoB and apoE in HepG2 cells, indicating that the effects of PAF or Lexipafant on the apoM production on hepatic cells are selective for apoM. The cellular mechanism of the effects of PAF or Lexipafant on apoM metabolism requires further investigations. PMID- 11944907 TI - Differential regulation of neutrophil phospholipase d activity and degranulation. AB - One of the proposed functions of phosphatidic acid (PA) formation from phospholipase D (PLD) activation in neutrophils is to promote degranulation induced by receptor agonists. The present study shows that the time course and dose response of PA formation and degranulation induced by N-formyl-methionyl leucyl-phenylalanine (fMLP) differed. PLD activation and degranulation also exhibited different dose response to genistein and epigallocatechin gallate (EGCG), inhibitors of protein tyrosine kinases. Genistein inhibited PLD activity with an IC(50) value of 12.2 microM in fMLP- and 107 microM in phorbol myristate acetate (PMA)-stimulated cells. It required higher concentrations of genistein to inhibit degranulation than to inhibit PLD activity induced by fMLP. EGCG in the range of 40-400 microM had no effect on PLD activity but it inhibited the release of beta-glucuronidase and elastase by fMLP-stimulated cells. These results demonstrate differential regulation of PLD activity and degranulation of primary granules by genistein and EGCG in fMLP-stimulated neutrophils. PMID- 11944908 TI - Cytoplasmic TSC-22 (transforming growth factor-beta-stimulated clone-22) markedly enhances the radiation sensitivity of salivary gland cancer cells. AB - We transfected a salivary gland cancer cell line, TYS, with three different forms of TSC-22 (transforming growth factor-beta-stimulated clone-22) gene: full-length TSC-22 (TSC-22FL) containing nuclear export signal, TSC-box and leucine zipper, truncated TSC-22 (TSC-22LZ) containing only TSC-box and leucine zipper, and truncated TSC-22 with nuclear localization signal (NLS-TSC-22LZ). High expression of TSC-22FL in the cytoplasm markedly enhanced the radiation-sensitivity of TYS cells, while, moderate expression of TSC-22FL marginally affected the radiation sensitivity. TSC-22LZ, which was expressed in the cytoplasm and the nucleus, enhanced the radiation-sensitivity of TYS cells irrespective to its expression level. NLS-TSC-22LZ, which was expressed only in the nucleus, marginally affected the radiation-sensitivity of the cells even at high expression level. Interestingly, cytoplasmic TSC-22 translocates to nucleus concomitant with radiation-induced apoptosis. These results suggest that cytoplasmic localization of TSC-22 and translocation of TSC-22 from cytoplasm to nucleus is important for regulating the cell death signal after irradiation-induced DNA damage. PMID- 11944909 TI - L-Methionine-gamma-lyase, as a target to inhibit malodorous bacterial growth by trifluoromethionine. AB - Methyl mercaptan is a major component responsible for oral malodor. This compound arises from the bacterial metabolism of methionine. Here we show that the growth of Porphyromonas gingivalis, a periodontal microorganism that produces large amounts of methyl mercaptan, was strongly inhibited by l-trifluoromethionine (TFM), a fluorinated derivative of methionine. In contrast, TFM had no effect on the growth of bacteria which do not produce methyl mercaptan. In addition, the survival rate of P. gingivalis-infected mice was remarkably increased by the co injection of TFM. These results suggest that TFM is a promising antibacterial agent specific to the malodorous oral bacteria. PMID- 11944910 TI - Transcriptional regulation of mouse MARCKS promoter in immortalized hippocampal cells. AB - Mouse MARCKS is a prominent myristoylated alanine-rich C kinase substrate implicated in brain development, calcium/calmodulin signaling, and membrane cytoskeletal restructuring, and is developmentally regulated in a cell- and tissue-specific fashion. In this study, transcriptional regulation of mouse MARCKS promoter in the neuronally derived immortalized hippocampal cells (HN33) was examined for a portion of 5'-flanking genomic sequence from -993 to +1 relative to the translation start site. Transfection experiments carried out in this neural cell line identified, for the first time, that the distal promoter segment from -993 to -713 plays a crucial role as an enhancer/activator element in the up-regulation of the basal transcription activity driven by MARCKS core promoter sequence. Motif analyses revealed at least 12 overlapping potential transcription factor binding sites in this region, among which a prominent GA rich sequence centered at -765 has been shown to be functionally important in the binding of Sp1 protein-like complex. Deletion of the GA-rich segment significantly reduced the MARCKS promoter activity. Further, competitive EMSA indicated two additional sites within the -993/-713 that may also interact with Sp1 protein, demonstrating that the activator function of -993/-713 is under control of multiple Sp1 transcription factors. Unlike the distal promoter sequence, the proximal core promoter sequence (-649/-438) contains a GC-rich box and a Z-DNA-forming segment and is critical to basal transcription. The deletion of -649/-438 segment has been shown to drastically impair the promoter activity even in the presence of -993/-713, suggesting that its presence is also important to the function of -993/-713. These data emphasize that the synergistic interaction between distal and proximal promoter sequences is indispensable for the optimal MARCKS promoter function in the immortalized hippocampal cells. The discovery of the activator function of the MARCKS distal promoter region, and its potential interaction with multiple Sp proteins may provide a new clue to the understanding of Macs transcriptional regulation in brain. PMID- 11944911 TI - Visualization of sequential exocytosis in rat pancreatic islet beta cells. AB - The examination of insulin exocytosis at the single cell level by conventional electrophysiologic and amperometric methods possesses inherent limitations, and may not accurately reflect the morphologic events of exocytosis of the insulin granule. To overcome some of these limitations, we show by epifluorescent microscopy of a fluorescent dye, FM1-43, its incorporation into the plasma membrane and oncoming insulin granules undergoing exocytosis, and their core proteins. Using this method, we tracked exocytosis in real-time in insulinoma INS 1 and single rat islet beta cells in response to KCl and glucose. We observed both single transient and multi-stepwise increases in membrane FM1-43 fluorescence, suggesting single granule exocytosis as well as sequential and compound exocytosis, respectively. Confocal microscopy of nonpermeabilized cells shows that some of the exocytosed insulin granules labeled by the FM1-43 dye could also be labeled with insulin antibodies, suggesting prolonged openings of the fusion pores and slow dissolution of the granule core proteins on the membrane surface. PMID- 11944912 TI - Thrombopoietin induces p-selectin expression on platelets and subsequent platelet/leukocyte interactions. AB - Ligation of thrombopoietin (TPO) to the platelet c-Mpl receptor induces numerous biochemical pathways in the absence of aggregation. Two forms of recombinant TPO are currently in clinical trials for the treatment of thrombocytopenia. This study focuses on the effects of the full-length recombinant human TPO (rhTPO) on platelets in a whole blood system. Platelet-leukocyte associations (PLAs) were visualized following rhTPO stimulation as CD42b/CD 45 double positive clusters by FACS analysis. Treatment of washed platelets with rhTPO induced granule release and expression of the leukocyte adhesion receptor P-selectin (CD 62P) in the absence of aggregation and calcium mobilization. RhTPO also induced platelet leukocyte interactions in whole blood. Following stimulation, leukocytes were recruited by platelets through P-selectin in a calcium-dependent manner. rhTPO stimulates platelet-leukocyte associations in whole blood through expression of platelet P-selectin. To our knowledge, this is the first report that identifies TPO as a promoter of platelet-leukocyte interactions. PMID- 11944913 TI - Identification of a novel protein p59(scr), which is expressed at specific stages of mouse spermatogenesis. AB - A novel cDNA-encoding polypeptide of 545 amino acid residues was identified from a mouse testis cDNA library. The transcript of this gene, p59(scr), was predominantly expressed in the testis and was developmentally regulated during spermatogenesis. The encoded protein was expressed in spermatocytes and round spermatids within seminiferous tubules of the adult testis. Using an adult-mouse model of experimental unilateral cryptorchidism, it was observed that the expression of the p59(scr) mRNA was reduced in the cryptorchid testes in association with destruction of spermatogenesis. In vitro heat stress experiment further demonstrated that p59(scr) mRNA was more sensitive to heat stress than the other mRNA species, such as germ-cell-specific meiosis-activating kinase (mak) gene and a housekeeping beta-actin gene. Our results suggest that this novel p59(scr) gene is involved in spermatogenesis and may play an important role in development of testicular germ cells. PMID- 11944914 TI - Transgenic expression of the EXT2 gene in developing chondrocytes enhances the synthesis of heparan sulfate and bone formation in mice. AB - Hereditary multiple exostoses (HME), a dominantly inherited disorder characterized by multiple cartilaginous tumors, is caused by mutations in the gene for, EXT1 or EXT2. Recent studies have revealed that EXT1 and EXT2 are required for the biosynthesis of heparan sulfate and exert maximal transferase activity as a complex. The Drosophila homologue of EXT1 (tout-velu) regulates the movement and signaling of Hedgehog protein, which plays an important role in the regulation of chondrocyte differentiation and bone development. In this study, to investigate the biological role of EXT2 in bone development in vivo and the pathological role of HME mutations in the development of exostoses, we generated transgenic mice expressing EXT2 or mutant EXT2 in developing chondrocytes. Histological analyses and micro-CT scanning showed that the biosynthesis of heparan sulfate and the formation of trabeculae were upregulated in EXT2 transgenic mice, but not in mutant EXT2-transgenic mice. The expression of EXT1 is concomitantly upregulated in EXT2-transgenic and even mutant EXT2-transgenic mice, suggesting an interactive regulation of EXT1 and EXT2 expression. These findings support that the EXT2 gene encodes an essential component of the glycosyltransferase complex required for the biosynthesis of heparan sulfate, which may eventually modulate the signaling involved in bone formation. PMID- 11944915 TI - The effect of antioxidants on glycated albumin-induced cytotoxicity in bovine retinal pericytes. AB - Loss of retinal pericytes is the initial deficit in the early stage of diabetic retinopathy. Glycated albumin (GA) forms under hyperglycemic conditions and exists in the retinal blood vessels of diabetic patients with retinopathy. In this study, using a 3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyl tetrazolium bromide (MTT) reduction test, we investigated whether GA induces cytotoxicity in cultured bovine retinal pericytes, and whether the antioxidants, l-ascorbic acid, Trolox, and probucol, provide any protection from GA-mediated cytotoxicity. GA induced pericyte death in a dose-dependent manner. With increasing time, GA-induced cytotoxicity also increased despite no strict time dependence. Furthermore, this cell death was found to be mediated both by apoptosis, which was confirmed by apoptosis-specific fluorescent staining of nuclei and cell membranes, and mitochondrial damage, as elucidated by electron microscopy. All three antioxidants used in this study partially protected against GA-induced pericyte death, suggesting that oxidative stress plays a role in GA-induced pericyte death. The results indicate that GA induces cell death in cultured bovine retinal pericytes, and that certain antioxidants may reduce this cytotoxicity. PMID- 11944916 TI - Human retinal pigment epithelial cells express scavenger receptors BI and BII. AB - The retinal pigment epithelial (RPE) cells that underlie the retina ingest and metabolize thousands of lipid-rich photoreceptor outer segments (POS) every day. The scavenger receptor (SR) CD36 and integrin alphavbeta5 have been shown to participate in POS binding and internalization by RPE cells. The objective of the current study was to determine whether RPE cells express SRs other than CD36. Primary cultures of human RPE cells express both mRNA and protein for SR-BI and SR- BII. SR-BI and SR-BII mRNAs were detected by reverse transcription-polymerase chain reaction. SR-BI protein was detected by immune precipitation of [(35)S]methionine-labeled crude cellular extracts. SR-BII was detected by Western blotting of immune precipitated crude cellular extracts. SR-BI and SR-BII proteins were also detected by immunofluorescence staining of RPE cells in culture. The results suggest that these SRs may play a role in POS lipid binding and uptake by RPE cells in the eye. PMID- 11944917 TI - Interaction of S100 proteins with the antiallergic drugs, olopatadine, amlexanox, and cromolyn: identification of putative drug binding sites on S100A1 protein. AB - S100 proteins are a multigenic family of low-molecular-weight Ca(2+)-binding proteins comprising 19 members. These proteins undergo a conformational change by Ca(2+)-binding and consequently interact with their target proteins. Recently, we reported that two antiallergic drugs, Amlexanox and Cromolyn, bind to S100A12 and S100A13 of the S100 protein family. In the present study, we used a newly developed antiallergic drug, Olopatadine, as a ligand for affinity chromatography and examined binding specificity of the drug to S100 protein family. Olopatadine binds specifically to S100 proteins, such as S100A1, S100B, S100L, S100A12, and S100A13, in a Ca(2+)-dependent manner but not to calmodulin. Mutagenesis study showed that amino acid residues 76-85 in S100A1 are necessary for its binding to Olopatadine. In contrast, residues 89-94 were identified as an Amlexanox-binding site in S100A1. Moreover, Olopatadine did not competitively inhibit S100A1 binding site of Amlexanox. Furthermore, we showed that Olopatadine inhibited the binding of S100A1 target protein's binding site peptides to S100A1. These results indicate that C-terminal region of S100A1 is important for antiallergic drug binding, although the drug binding sites are different according to each antiallergic drug. Differences in the binding sites of S100A1 to antiallergic drugs suggest that the regulatory functions of S100 proteins may exist in several regions. Therefore, these drugs may serve as useful tools for evaluating the physiological significance of S100 protein family. PMID- 11944918 TI - Ghrelin, macronutrient intake and dietary preferences in long-evans rats. AB - Ghrelin is a recently discovered peptide that is primarily produced by the stomach. As a ligand of the growth hormone (GH) secretagogue (GHS) receptor, it stimulates GH secretion but it also stimulates feeding and has adipogenic effects in rodents. Although its circulating levels are modulated by fasting and refeeding, its relationship with diet composition is not known. In the present paper, we measured plasma ghrelin as well as two important hormones (leptin and insulin) in Long-Evans rats placed in two different feeding situations, e.g., either with imposed diets or with food choice. In the first case, the rats were fed unbalanced diets (either high-carbohydrate (HC) or high-fat (HF) diets) for 14 weeks, whereas in the second situation, they had the choice between these 2 diets for 2 weeks and were selected for their fat or carbohydrate preference. The intake of the HF diet for 14 weeks was associated with lower levels of ghrelin ( 30% vs control diet; P < 0.01). These levels increased when the percentage of carbohydrate in the diet increased (+26 to +42% vs control diet; P < 0.01 or less). Ghrelin was inversely correlated with plasma leptin (r = -0.55; P < 0.003) and blood glucose (r = -0.58; P < 0.001) as well as with body weight (r = -0.63; P < 0.0001) and body fat content estimated by the sampling of specific fat pads (r = -0.62; P < 0.0001). In the food choice experiment, fat-preferring rats had plasma ghrelin levels lower than the carbohydrate-preferring rats (-33%; P < 0.0002). Ghrelin secretion was therefore very sensitive to the diet composition. Its down-regulation by fat ingestion might serve as a counterregulatory mechanism to limit the development of dietary-induced adiposity. Ghrelin may signal when a high calorie diet is ingested. PMID- 11944919 TI - Discovery of a stable dimeric mutant of cyanovirin-N (CV-N) from a T7 phage displayed CV-N mutant library. AB - Mutant proteins with altered properties can be useful probes for investigating structure, ligand binding sites, mechanisms of action, and physicochemical attributes of the corresponding wild-type proteins of interest. In this report, we illuminate properties of mutants of the potent HIV-inactivating protein, cyanovirin-N (CV-N), selected by construction of a mutant library by error-prone polymerase chain reaction and affinity-based screening using T7 phage display technology. After three rounds of biopanning, two phage-displayed, one-point mutants of CV-N, Ser52Pro and Ala77Thr, were isolated. After the elucidation of biological activities of the mutants displayed on phage as well as the Escherichia coli-expressed, purified mutant proteins, we subsequently subjected the mutants to analyses by native PAGE and size-exclusion chromatography. We found that the Ser52Pro mutant not only was active against HIV but also existed exclusively as a dimer in solution. This was in marked contrast to the wild-type CV-N, which exists in solution predominantly as the monomer. The Ser52Pro mutant provides a novel model for further investigations of the folding mechanism as well as structure-activity requirements for CV-N's antiviral properties. PMID- 11944920 TI - Detection of a novel variant human hemoglobin by electrospray ionization mass spectrometry. AB - A novel hemoglobin variant was detected by electrospray ionization mass spectrometry. Hb Zurich-Hottingen is characterized by an Asn --> Ser replacement in the alpha-chain at position 9 as confirmed by DNA analysis. This hemoglobin variant is silent in isoelectric focusing, reversed-phase chromatography, and cation-exchange chromatography. The mutant alpha-chain was detectable only with electrospray mass spectrometry by its mass shift of -27 Da. The carrier was found to be heterozygous for the new hemoglobin variant. These results illustrate the power of ESI mass spectrometry for hemoglobin analysis. PMID- 11944921 TI - The human monocarboxylate transporter, MCT1: genomic organization and promoter analysis. AB - Uptake of butyrate across the colonocyte luminal membrane is mediated by the monocarboxylate transporter isoform 1 (MCT1). We have demonstrated previously that expression of human colonic MCT1 is responsive to butyrate, and that this involves the dual control of MCT1 gene transcription and stability of the MCT1 transcript. Here we describe the structural organization of the human MCT1 gene, and report the isolation and characterization of the MCT1 gene promoter. The MCT1 gene spans approximately 44 kb, and is organized as 5 exons intervened by 4 introns. The first of these introns is located in the 5'-UTR-encoding DNA, spans >26 kb, and thus accounts for approximately 60% of the entire transcription unit. Analysis of a 1.5 kb fragment of the MCT1 5'-flanking region, shows an absence of the classical TATA-Box motif. However, the region contains potential binding sites for a variety of transcription factors with known association with butyrate's action in the colon. In transient transfections the 5'-flanking region drives high-level expression of a luciferase reporter-gene in cells that endogenously express MCT1. Deletion analyses indicate that the cis-acting elements necessary for basal transcription of MCT1 are contained within the 70/+213 proximal sequence of the promoter. PMID- 11944922 TI - BAY 41-2272 activates two isoforms of nitric oxide-sensitive guanylyl cyclase. AB - Soluble guanylyl cyclase is an important target for endogenous nitric oxide and the guanylyl cyclase modulator, YC-1. Recently BAY 41-2272 was identified as a similar but more potent and more specific substance. While YC-1 also acts as non specific phosphodiesterase inhibitor, BAY 41-2272 is devoid of an effect on phosphodiesterases. BAY 41-2272 has so far only been tested on the alpha(1)/beta(1) heterodimeric isoform of soluble guanylyl cyclase and its binding site has been mapped to a region in the alpha(1) subunit amino-terminal sequence. Although this region is poorly conserved in the alpha(2) subunit, we show in the current study that the alpha(2)/beta(1) heterodimeric enzyme isoform is activated by BAY 41-2272. Deletion analysis of the alpha(2) subunit and co expression with the beta(1) subunit in the baculovirus/Sf9 system is consistent with the amino-terminal amino acids 104 to 401 of the alpha(2) subunit as binding site for BAY 41-2272. PMID- 11944923 TI - Species-specific and isoform-specific RNA binding of human and mouse fragile X mental retardation proteins. AB - The loss of the fragile X RNA binding protein, FMRP, causes macroorchidism and mental retardation in man. The discovery of a mouse ortholog led to the development of several FMRP knockout mouse strains that recapitulate some features of the disease. As mouse and human FMRPs differ in several amino acids in their RNA binding domains, we compared the RNA binding profiles of these two orthologs. Five variant FMRPs, whose differences arose from alternative splicing and mutation within the conserved RNA binding domains, were examined. Homoribopolymer binding studies showed that human FMRPs (hFMRP) bound a broader range of single-stranded mimetics than mouse FMRPs (mFMRP) and these interactions were both complex and cooperative. hFMRP and mFMRP also displayed significant preferences toward binding their own mRNA; specifically we found that the mFMRP isoforms bind mFMR1 mRNA much more tightly than their human counterparts. Finally, these data demonstrate that each FMRP variant binds RNAs uniquely, resulting in a set of proteins with differing affinities. PMID- 11944924 TI - Identification and fine mapping of IgG and IgE epitopes in ovomucoid. AB - Ovomucoid is a major allergen in hen egg white which causes a serious IgE mediated food allergy reaction. This study determined eight IgG epitopes, 5-11 amino acids in length, and nine IgE epitopes, 5-16 amino acids in length, within the primary sequence in ovomucoid using arrays of overlapping peptides synthesized on cellulose membranes. Pooled sera from eight egg-allergic patients were used to probe the membrane. We also analyzed the amino acids that are critical for antibody binding by substituting a single amino acid within each epitope. Mutational analysis of the epitopes indicated that charged amino acids (aspartic acid, glutamic acid, and lysine) and some hydrophobic (leucine, phenylalanine, and glycine) and polar (serine, threonine, tyrosine, and cystein) amino acids were important for antibody binding. These results provide useful information for the molecular design necessary to reduce the allergenicity of ovomucoid, and a better understanding of structure-function relationships of allergic epitopes in food proteins. PMID- 11944925 TI - Design and synthesis of highly potent and selective melanotropin analogues of SHU9119 modified at position 6. AB - The melanocortin receptors are involved in several important physiological functions. The potent and enzymatically stable analogues MT-II (Ac-Nle-c[Asp-His DPhe-Arg-Trp-Lys]-NH(2)) and SHU9119 (Ac-Nle-c[Asp-His-DNal(2')-Arg-Trp-Lys] NH(2)) are important ligands of these receptors but are relatively nonselective. To differentiate between the physiological functions of these receptors, agonists, and antagonists with improved receptor selectivities are needed. We report here analogues of the well-characterized antagonist SHU9119 in which we replaced His(6) with conformationally constrained amino acids. By this structure activity study we discovered two important compounds, PG-901 (Ac-Nle(4)-c[Asp(5) Pro(6)-DNal(2')(7)-Arg(8)-Trp(9)-Lys(10)]-NH(2)) and PG-911 (Ac-Nle(4)-c[Asp(5) Hyp(6)-DNal(2')(7)-Arg(8)-Trp(9)-Lys(10)]-NH(2)), characterized to be full agonists at the hMC5R (EC(50) = 0.072 nM and 0.031 nM, respectively), but full antagonists at the hMC3R and the hMC4R. We also demonstrated that the relative stereochemistry of the amino acid at the 6-position is critical for activity, and could play an important role in potency as well as in selectivity for the melanocortin receptors. PMID- 11944926 TI - Antimorphic PV.1 causes secondary axis by inducing ectopic organizer. AB - Xenopus homeobox gene, PV.1 ventralizes activin-induced dorsal mesoderm and inhibits neuralization of ectoderm in animal cap when overexpressed. Here we generated PV.1/engrailed fusion construct (N-PV1-EnR) to perform loss-of-function study for this transcription factor. N-PV1-EnR showed an extremely antimorphic effect, causing a partial secondary embryonic axis when expressed at ventral marginal zone of blastula. In ventral marginal zone cells, this chimeric protein induced organizer genes and suppressed ventral markers mimicking those effects reported for dominant negative BMP-4 receptor (DNBR). Moreover, N-PV1-EnR rescued the ventralized embryos caused by the ectopic dorsal expression of PV.1 but not by that of Xvent-2. These results suggested that PV.1 functions at downstream of BMP-4 as a ventralizing effector which acts separately from Xvent-2 and the dominant negative effect gained by this specific mutant is applicable for the further studies of BMP-4 downstream pathway. PMID- 11944927 TI - Potent and selective partial ecdysone agonist activity of chromafenozide in Sf9 cells. AB - Chromafenozide (ANS-118) is a non-steroidal ecdysone mimic and its insecticidal effect is highly specific to lepidoptera. In order to evaluate the transcription inducing activity via nuclear ecdysone receptor (EcR) and the mode of action of chromafenozide, ecdysone-responsive reporter gene assay systems were developed in Sf9 and Kc cells. Ponasterone A, a full EcR agonist, induced reporter transcription in a dose-dependent manner in both Sf9 and Kc cells. In contrast, chromafenozide activated reporter transcription with comparable potency to ponasterone A only in Sf9 cells, although its maximum activity was 4-fold lower than that of ponasterone A. When chromafenozide was applied together with ponasterone A to Sf9 cells, it antagonized ponasterone A at nanomolar concentrations. These results suggest that chromafenozide is a potent partial EcR agonist specific to lepidoptera; it appears to bind lepidopteran EcR with comparable affinity to ponasterone A, but may activate the EcR in a different manner. PMID- 11944928 TI - The adhesion receptor CD-31 can be primed to rapidly adjust the neutrophil cytoskeleton. AB - The adhesion receptor CD-31 is expressed on neutrophils and endothelial cells and participates in transendothelial migration of neutrophils. Although necessary, information on CD-31-induced signaling and its influence on the shape-forming actin network is scarce. Here, we found that antibody engagement of CD-31 on suspended neutrophils triggered a prompt intracellular Ca(2+) signal, providing the cells had been primed with a chemotactic factor. Inhibition of Src-tyrosine kinases blocked this Ca(2+) signal, but not a fMet-Leu-Phe-induced Ca(2+) signal. Despite the ability of fMet-Leu-Phe to activate Src-tyrosine kinases, it did not per se induce tyrosine phosphorylation of CD-31. However, fMet-Leu-Phe did enable such a phosphorylation following an antibody-induced engagement of CD-31. This clustering also triggered a Ca(2+)-dependent depolymerization of actin and, surprisingly enough, a simultaneous polymerization. The ability of CD-31 to signal dynamic alterations in the cytoskeleton, particularly the Ca(2+)-induced actin depolymerization, further explains how neutrophils can squeeze themselves out between adjacent endothelial cells. PMID- 11944929 TI - p85 binds to G-actin in a Ca(2+)/calmodulin-dependent manner, thus regulating the initiation of cytokinesis in tetrahymena. AB - Tetrahymena p85 is localized to the presumptive division plane before the formation of contractile ring microfilaments. p85 binds to calmodulin in a Ca(2+) dependent manner and both proteins colocalize to the division furrow. Inhibition of the binding of p85 and Ca(2+)/calmodulin prevents both the localization of p85 and calmodulin to the division plane and the formation of the contractile ring, suggesting that the interaction of p85 and Ca(2+)/calmodulin is important in the formation of the contractile ring. We investigated the mechanisms of the formation of contractile ring, and the relationship among p85, CaM, and actin using co-sedimentation assay: p85 binds to G-actin in a Ca(2+)/calmodulin dependent manner, but does not bind to F-actin. Therefore, we propose that a Ca(2+)/calmodulin signal and its target protein p85 are cooperatively involved in the recruitment of G-actin to the division plane and the formation of the contractile ring. PMID- 11944930 TI - Synthesis and characterization of N-coumaroyltyramine as a potent phytochemical which arrests human transformed cells via inhibiting protein tyrosine kinases. AB - Numerous phytochemicals are believed to have beneficial effects on human health. N-Coumaroyltyramine accumulates in plants in response to wounding and pathogen attack. Due to the scarcity of N-coumaroyltyramine, its biological activities have not been studied in human cells. In this study, N-coumaroyltyramine was chemically synthesized and then purified by an HPLC with a UV-visible absorbance detector. Retention times of major peaks were 14.3 and 20.7 min, and the peak at 20.7 min was confirmed by LC-MS as N-coumaroyltyramine with a mass/charge (m/z) unit of 284.1. The synthesis procedure was relatively easy and had an acceptable yield (approximately 55%). The compound exhibited a new activity, suppression of growth of human tumor cells such as U937 and Jurkat cells. In addition, the suppressed growth of the cells was strongly associated with an increased percentage of cells in the S phase of the cell cycle progression. Furthermore, N coumaroyltyramine was able to inhibit the protein tyrosine kinases including epidermal growth factor receptor (EGFR). This is the first report of the growth suppressing activity of N-coumaroyltyramine and its arrest of cells at the S phase of the cell cycle, possibly by inhibition of protein tyrosine kinases. PMID- 11944931 TI - A circular RNA-DNA enzyme obtained by in vitro selection. AB - A circular RNA-DNA enzyme with higher activity to target RNA cleavage and higher stability than that of the hammerhead ribozyme in the presence of RNase A was obtained by in vitro selection. The molecule is composed of a catalytic domain of 22-mer ribonucleotides derived from the hammerhead ribozyme and a fragment of 55 mer deoxyribonucleotides. The DNA fragment contains two substrate-binding domains (9-mer and 6-mer, respectively) and a "regulation domain" (assistant 40-mer DNA with 20-mer random deoxyribonucleotides sequence), which probably play the role in the regulation of flexibility and rigidity of the circular RNA-DNA enzyme. The above results suggest that the circular RNA-DNA enzyme will have a great prospect in gene-targeting therapies. PMID- 11944932 TI - Differential effects of specific phosphodiesterase isoenzyme inhibitors on bovine oocyte meiotic maturation. AB - The differential regulation of cAMP levels within the oocyte and somatic (cumulus) cell compartments of the bovine follicle, and the subsequent regulation of oocyte meiotic maturation was examined through specific cell-type localisation of phosphodiesterases (PDEs). Selective PDE inhibitors were used to modulate cAMP levels in each of the two follicular compartments and to examine their effects on oocyte meiotic maturation. Ovaries were obtained from an abattoir and cumulus oocyte complexes (COC) were aspirated from antral follicles into culture medium supplemented with 4 mg/ml BSA and 2mM 3-isobutyl-1-methylxanthine (IBMX). COC, denuded oocytes (DO), or mural granulosa cells (MGC) were cultured either with or without forskolin or FSH, in the presence of specific PDE inhibitors; either milrinone (PDE3 inhibitor), cilostamide (PDE3 inhibitor), or rolipram (PDE4 inhibitor). COC/DO cultures were assessed for meiotic progression and cAMP content, and MGC for cAMP production. The type 3 PDE inhibitor, but not the type 4, prevented spontaneous meiotic maturation and elevated intraoocyte cAMP in cultured denuded oocytes. In contrast, the type 4 PDE inhibitor had no effect on the oocyte, but elevated mural granulosa and cumulus cell cAMP production. The results of this study indicate that specific PDE subtypes are differentially localised within the two compartments of the bovine follicle-the type 3 PDE in the oocyte and the type 4 PDE in the granulosa cells. In addition, oocyte cAMP levels are primarily regulated in bovine oocytes by its degradation by PDE, whereas granulosa cell cAMP levels are controlled mainly by active adenylate cyclase, with both sources able to participate in oocyte meiotic regulation. PMID- 11944933 TI - Reciprocal regulatory interactions between the Notch and Ras signaling pathways in the Drosophila embryonic mesoderm. AB - Convergent intercellular signals must be precisely integrated in order to elicit specific biological responses. During specification of muscle and cardiac progenitors from clusters of equivalent cells in the Drosophila embryonic mesoderm, the Ras/MAPK pathway--activated by both epidermal and fibroblast growth factor receptors--functions as an inductive cellular determination signal, while lateral inhibition mediated by Notch antagonizes this activity. A critical balance between these signals must be achieved to enable one cell of an equivalence group to segregate as a progenitor while its neighbors assume a nonprogenitor identity. We have investigated whether these opposing signals directly interact with each other, and we have examined how they are integrated by the responding cells to specify their unique fates. Our findings reveal that Ras and Notch do not function independently; rather, we have uncovered several modes of cross-talk between these pathways. Ras induces Notch, its ligand Delta, and the epidermal growth factor receptor antagonist, Argos. We show that Delta and Argos then synergize to nonautonomously block a positive autoregulatory feedback loop that amplifies a fate-inducing Ras signal. This feedback loop is characterized by Ras-mediated upregulation of proximal components of both the epidermal and fibroblast growth factor receptor pathways. In turn, Notch activation in nonprogenitors induces its own expression and simultaneously suppresses both Delta and Argos levels, thereby reinforcing a unidirectional inhibitory response. These reciprocal interactions combine to generate the signal thresholds that are essential for proper specification of progenitors and nonprogenitors from groups of initially equivalent cells. PMID- 11944934 TI - Nkx-2.5 gene induction in mice is mediated by a Smad consensus regulatory region. AB - In the forming vertebrate heart, bone morphogenetic protein signaling induces expression of the early cardiac regulatory gene nkx-2.5. A similar regulatory interaction has been defined in Drosophila embryos where Dpp signaling mediated by the Smad homologues Mad and Medea directly regulates early cardiac expression of tinman. A conserved cluster of Smad consensus binding sequences was identified in early cardiac regulatory sequences of the mouse nkx-2.5 gene. The importance of the nkx-2.5 Smad consensus region in early cardiac gene expression was examined in transgenic mice and in cultured mouse embryos. In transgenic mice, deletion of the Smad consensus region delays induction of embryonic DeltaSmadnkx 2.5/lacZ gene expression during early heart formation. Induction of DeltaSmadnkx 2.5/lacZ expression is also delayed in the outflow tract myocardium and visceral mesoderm. Targeted mutation of the three Smad consensus sequences inhibited nkx 2.5/lacZ expression in the cardiac crescent, demonstrating a specific requirement for the Smad consensus sites in early cardiac gene induction. Cultured DeltaSmadnkx-2.5/lacZ transgenic mouse embryos also exhibit delayed induction of transgene expression. In the four-chambered heart, deletion of the Smad consensus region resulted in expanded DeltaSmadnkx-2.5/lacZ transgene expression. Thus, the nkx-2.5 Smad consensus region can have positive or negative regulatory function, depending on the developmental context and cellular environment. PMID- 11944935 TI - Cardiac-specific activity of an Nkx2-5 enhancer requires an evolutionarily conserved Smad binding site. AB - Heart formation in vertebrates and fruit flies requires signaling by bone morphogenetic proteins (BMPs) to cardiogenic mesodermal precursor cells. The vertebrate homeobox gene Nkx2-5 and its Drosophila ortholog, tinman, are the earliest known markers for the cardiac lineage. Transcriptional activation of tinman expression in the cardiac lineage is dependent on a mesoderm-specific enhancer that binds Smad proteins, which activate transcription in response to BMP signaling, and Tinman, which maintains its own expression through an autoregulatory loop. Here, we show that an evolutionarily conserved, cardiac specific enhancer of the mouse Nkx2-5 gene contains multiple Smad binding sites, as well as a binding site for Nkx2-5. A single Smad site is required for enhancer activity at early and late stages of heart development in vivo, whereas the Nkx2 5 site is not required for enhancer activity. These findings demonstrate that Nkx2-5, like tinman, is a direct target for transcriptional activation by Smad proteins; however, the independence of this Nkx2-5 enhancer of Nkx2-5 binding suggests a fundamental difference in the transcriptional circuitry for activation of Nkx2-5 and tinman expression during cardiogenesis in vertebrates and fruit flies. PMID- 11944936 TI - Axial protocadherin is a mediator of prenotochord cell sorting in Xenopus. AB - Prenotochord cell sorting is regarded as one of the first cell sorting events in early chordate development. We recently demonstrated that this sorting event occurs in vitro, although the mediator of this activity remains unidentified. Herein, we report the isolation of a full-length cDNA clone of Axial protocadherin (AXPC), the homologue of human protocadherin-1 (PCD1). AXPC encodes a transmembrane protein (AXPC) that is expressed exclusively in the notochord at the neurula stage and in the pronephros, somites, heart, optic vesicle, otic vesicle, and distinct parts of the brain at the tailbud stage. Cell dissociation and reaggregation assays and in vivo microinjection experiments demonstrated that cells overexpressing a membrane-tethered form of AXPC (MT-AXPC) acquired the same adhesive properties as prenotochord cells. Moreover, microinjection of either mRNA encoding the dominant negative form of AXPC (DN-AXPC) or morpholino oligonucleotides interferes with the sorting activity of prenotochord cells and normal axis formation. This study suggests that AXPC is necessary and sufficient for prenotochord cell sorting in the gastrulating embryo, and may also mediate sorting events later in development. PMID- 11944937 TI - The development of three identified motor neurons in the larva of an ascidian, Halocynthia roretzi. AB - The generation of distinct classes of motor neurons underlies the development of complex motile behavior in all animals and is well characterized in chordates. Recent molecular studies indicate that the ascidian larval central nervous system (CNS) exhibits anteroposterior regionalization similar to that seen in the vertebrate CNS. To extend the understanding about the diversity of motor neurons in the ascidian larva, we have identified the number, position, and projection of individual motor neurons in Halocynthia roretzi, using a green fluorescent protein under the control of a neuron-specific promoter. Three pairs of motor neurons, each with a distinct shape and innervation pattern, were identified along the anteroposterior axis of the neural tube: the anterior and posterior pairs extend their axons toward dorsal muscle cells, whereas the middle pair project their axons toward ventral muscle. Overexpression of a dominant-negative form of a potassium channel in these cells resulted in paralysis on the injected side, thus these cells must constitute the major population of motor neurons responsible for swimming behavior. Lim class homeobox genes have been known as candidate genes that determine subtypes of motor neurons. Therefore, the expression pattern of Hrlim, which is a Lim class homeobox gene, was examined in the motor neuron precursors. All three motor neurons expressed Hrlim at the tailbud stage, although each down-regulated Hrlim at a different time. Misexpression of Hrlim in the epidermal lineage led to ectopic expression of TuNa2, a putative voltage-gated channel gene normally expressed predominantly in the three pairs of motor neurons. Hrlim may control membrane excitability of motor neurons by regulating ion channel gene expression. PMID- 11944938 TI - Regulation of synaptotagmin gene expression during ascidian embryogenesis. AB - The ascidian embryo, a model for the primitive mode of chordate development, rapidly forms a dorsal nervous system which consists of a small number of neurons. Here, we have characterized the transcriptional regulation of an ascidian synaptotagmin (syt) gene to explore the molecular mechanisms underlying development of synaptic transmission. In situ hybridization showed that syt is expressed in all neurons described in previous studies and transiently in the embryonic epidermis. Neuronal expression of syt requires induction from the vegetal side of the embryo, whereas epidermal expression occurs autonomously in isolated ectodermal blastomeres. Introduction of green fluorescent protein reporter gene constructs into the ascidian embryos indicates that a genomic fragment of the 3.4-kb 5' upstream region contains promoter elements of syt gene. Deletion analysis of the promoter suggests that syt expression in neurons and in the embryonic epidermis depends on distinct cis-regulatory regions. PMID- 11944939 TI - Efficient recombination in diverse tissues by a tamoxifen-inducible form of Cre: a tool for temporally regulated gene activation/inactivation in the mouse. AB - In recent years, the Cre integrase from bacteriophage P1 has become an essential tool for conditional gene activation and/or inactivation in mouse. In an earlier report, we described a fusion protein between Cre and a mutated form of the ligand binding domain of the estrogen receptor (Cre-ER) that renders Cre activity tamoxifen (TM) inducible, allowing for conditional modification of gene activity in the mammalian neural tube in utero. In the current work, we have generated a transgenic mouse line in which Cre-ER is ubiquitously expressed to permit temporally regulated Cre-mediated recombination in diverse tissues of the mouse at embryonic and adult stages. We demonstrate that a single, intraperitoneal injection of TM into a pregnant mouse at 8.5 days postcoitum leads to detectable recombination in the developing embryo within 6 h of injection and efficient recombination of a reporter gene in derivatives of all three germ layers within 24 h of injection. In addition, by varying the dose of TM injected, the percentage of cells undergoing a recombination event in the embryo can be controlled. Dose-dependent excision induced by TM was also possible in diverse tissues in the adult mouse, including the central nervous system, and in cultured cells derived from the transgenic mouse line. This inducible Cre system will be a broadly useful tool to modulate gene activity in mouse embryos, adults, and culture systems where temporal control is an important consideration. PMID- 11944940 TI - Complementary expression of IGF-II and IGFBP-5 during anterior pituitary development. AB - The specification of the five individual hormone-secreting cell types in the anterior pituitary requires a series of sequential cell fate decisions. We have immortalized cells at several stages along this pathway of pituitary differentiation. Here, we present analysis of differences in gene expression between an anterior pituitary precursor cell line, alphaT1-1, and an immature gonadotrope cell line, alphaT3-1, identified by using cDNA subtraction. Messenger RNA expression of members of the insulin-like growth factor signaling system, IGF II and IGFBP-5, was found in the alphaT1-1 precursor cell line, but not in the more differentiated cell line, alphaT3-1. This inferred stage specificity was confirmed in the mouse embryo by using in situ hybridization on embryonic days e10.5 through e18.5. Expression of IGF-II and IGFBP-5 mRNAs was both temporally and spatially regulated during pituitary development. IGF-II was highly expressed in the epithelium surrounding Rathke's pouch at e10.5, while IGFBP-5 expression was restricted to the adjacent oral epithelium. At e11.5 (represented by alphaT1 1), IGF-II was expressed throughout the pouch, but was coexpressed with IGFBP-5 and alpha-subunit in the ventral portion of the pouch epithelium. On e12.5, the two mRNAs were expressed in opposing dorsoventral (IGF-II) and ventrodorsal (IGFBP-5) patterns, with IGF-II excluded from the rostral, alpha-subunit expressing region. A decrease of both mRNAs was observed at e14.5 (equivalent to alphaT3-1), with IGF-II levels low and IGFBP-5 concentrated in the anterior pituitary rostral tip. These findings suggest that the timing of IGF-II expression and regulation of its accessibility by IGFBP-5 may play a role in anterior pituitary differentiation, survival, and/or proliferation. PMID- 11944941 TI - Zic1 promotes the expansion of dorsal neural progenitors in spinal cord by inhibiting neuronal differentiation. AB - The role of Zic1 was investigated by altering its expression status in developing spinal cords. Zic genes encode zinc finger proteins homologous to Drosophila Odd paired. In vertebrate neural development, they are generally expressed in the dorsal neural tube. Chick Zic1 was initially expressed evenly along the dorsoventral axis and its expression became increasingly restricted dorsally during the course of neurulation. The dorsal expression of Zic1 was regulated by Sonic hedgehog, BMP4, and BMP7, as revealed by their overexpressions in the spinal cord. When Zic1 was misexpressed on the ventral side of the chick spinal cord, neuronal differentiation was inhibited irrespective of the dorsoventral position. In addition, dorsoventral properties were not grossly affected as revealed by molecular markers. Concordantly, when Zic1 was overexpressed in the dorsal spinal cord in transgenic mice, we observed hypercellularity in the dorsal spinal cord. The transgene-expressing cells were increased in comparison to those of truncated mutant Zic1-bearing mice. Conversely, we observed a significant cell number reduction without loss of dorsal properties in the dorsal spinal cords of Zic1-deficient mice. Taken together, these findings suggest that Zic1 controls the expansion of neuronal precursors by inhibiting the progression of neuronal differentiation. Notch-mediated inhibition of neuronal differentiation is likely to act downstream of Zic genes since Notch1 is upregulated in Zic1-overexpressing spinal cords in both the mouse and the chick. PMID- 11944942 TI - Xenopus Cdc42 regulates convergent extension movements during gastrulation through Wnt/Ca2+ signaling pathway. AB - Rho GTPases are molecular switches that regulate many essential cellular processes, including actin dynamics, cell adhesion, cell-cycle progression, and transcription. We have isolated the Xenopus homolog of Rho GTPase Cdc42 and examined its potential role during gastrulation movements in early Xenopus embryos. XCdc42 is expressed in tissues undergoing extensive morphogenetic changes, such as the deep layers of involuting mesoderm and posterior neuroectoderm during gastrulation, and somitic mesoderm at neurula stages. Overexpression of either wild-type (WT) or dominant-negative (DN) XCdc42 interferes with convergent extension movements in intact embryos, activin stimulated animal caps, and dorsal marginal zone explants. These effects occur without affecting mesodermal specification. Overexpression of WT or DN XCdc42 leads to the decrease and increase of cell adhesiveness of blastomeres, respectively, as demonstrated by the cell adhesion assay. In addition, when overexpressed, PKC-alpha, XWnt-5a, and Mfz-3 inhibit activin-induced convergent extension in animal cap explants. This inhibition can be rescued by coexpression of DN XCdc42, implying that XCdc42 acts downstream of the Wnt/Ca2+ signaling pathway involving PKC activation. XCdc42 also lies downstream of XWnt-5a in the regulation of Ca2+-dependent cell adhesion. Taken together, our results suggest that XCdc42 plays a role in the regulation of convergent extension movements during gastrulation through the protein kinase C-mediated Wnt/Ca2+ pathway. PMID- 11944943 TI - Epithelia-mesenchyme interaction plays an essential role in transdifferentiation of retinal pigment epithelium of silver mutant quail: localization of FGF and related molecules and aberrant migration pattern of neural crest cells during eye rudiment formation. AB - Homozygotes of the quail silver mutation, which have plumage color changes, also display a unique phenotype in the eye: during early embryonic development, the retinal pigment epithelium (RPE) spontaneously transdifferentiates into neural retinal tissue. Mitf is considered to be the responsible gene and to function similarly to the mouse microphthalmia mutation, and tissue interaction between RPE and surrounding mesenchymal tissue in organ culture has been shown to be essential for the initiation of the transdifferentiation process in which fibroblast growth factor (FGF) signaling is involved. The immunohistochemical results of the present study show that laminin and heparan sulfate proteoglycan, both acting as cofactors for FGF binding, are localized in the area of transdifferentiation of silver embryos much more abundantly than in wild-type embryos. More intense immunohistochemical staining with FGF-1 antibody, but not with FGF-2 antibody, is also found in the neural retina, RPE, and choroidal tissue of silver embryos than in wild-type embryos. HNK-1 immunohistochemistry revealed that clusters of HNK-1-positive cells (presumptive migrating neural crest cells) are frequently located around the developing eyes and in the posterior region of the silver embryonic eye. Finally, chick-quail chimerical eyes were made by grafting silver quail optic vesicles to chicken host embryos: in most cases, no transdifferentiation occurs in the silver RPE, but in a few cases, transdifferentiation occurs where silver quail cells predominate in the choroid tissue. These observations together with our previous in vitro study indicate that the silver mutation affects not only RPE cells but also cephalic neural crest cells, which migrate to the eye rudiment, and that these crest cells play an essential role in the transdifferentiation of RPE, possibly by modifying the FGF signaling pathway. The precise molecular mechanism involved in RPE-neural crest cell interaction is still unknown, and the quail silver mutation is considered to be a good experimental model for studying the role of neural crest cells in vertebrate eye development. PMID- 11944944 TI - Conservation of Brachyury, Mef2, and Snail in the myogenic lineage of jellyfish: a connection to the mesoderm of bilateria. AB - One major difference between simple metazoans such as cnidarians and all the bilaterian animals is thought to involve the invention of mesoderm. The terms diploblasts and triploblasts are therefore, often used to group prebilaterian and bilaterian animals, respectively. However, jellyfish contain well developed striated and smooth muscle tissues that derive from the entocodon, a mesoderm like tissue formed during medusa development. We investigated the hypothesis, that the entocodon could be homologous to the third germ layer of bilaterians by analyzing the structures and expression patterns of the homologues of Brachyury, Mef2, and Snail in the jellyfish Podocoryne carnea. These are regulatory genes from the T-box, MADS-box and zinc finger families known to play important roles in bilaterian mesoderm patterning and muscle differentiation. The sequence and expression data demonstrate that the genes are structurally and functionally conserved and even more similar to humans or other deuterostomes than to protostome model organisms such as Drosophila or Caenorhabditis elegans. Based on these data we conclude that the common ancestor of the cnidarians and bilaterians not only shared genes that play a role in regulating myogenesis but already used them to develop and differentiate muscle systems similar to those of triploblasts. PMID- 11944945 TI - Tail morphogenesis in the ascidian, Ciona intestinalis, requires cooperation between notochord and muscle. AB - We present evidence that notochord and muscle differentiation are crucial for morphogenesis of the ascidian tail. We developed a novel approach for embryological manipulation of the developing larval tissues using a simple method to introduce DNA into Ciona intestinalis and the several available tissue specific promoters. With such promoters, we misexpressed the Xenopus homeobox gene bix in notochord or muscle of Ciona embryos as a means of interfering with development of these tissues. Ciona embryos expressing bix in the notochord from the 64-cell stage develop into larvae with very short tails, in which the notochord precursors fail to intercalate and differentiate. Larvae with mosaic expression of bix have intermediate phenotypes, in which a partial notochord is formed by the precursor cells that did not receive the transgene while the precursors that express the transgene cluster together and fail to undergo any of the cell-shape changes associated with notochord differentiation. Muscle cells adjacent to differentiated notochord cells are properly patterned, while those next to the notochord precursor cells transformed by bix exhibit various patterning defects. In these embryos, the neural tube extends in the tail to form a nerve cord, while the endodermal strand fails to enter the tail region. Similarly, expression of bix in muscle progenitors impairs differentiation of muscle cells, and as a result, notochord cells fail to undergo normal extension movements. Hence, these larvae have a shorter tail, due to a block in the elongation of the notochord. Taken together, these observations suggest that tail formation in ascidian larvae requires not only signaling from notochord to muscle cells, but also a "retrograde" signal from muscle cells to notochord. PMID- 11944946 TI - Dcas is required for importin-alpha3 nuclear export and mechano-sensory organ cell fate specification in Drosophila. AB - We have studied the in vivo function and tissue specificity of Dcas, the Drosophila ortholog of CAS, the importin beta-like export receptor for importin alpha. While dcas mRNA is specifically expressed in the embryonic central nervous system, Dcas protein is maternally supplied to all embryonic cells and its nuclear/cytoplasmic distribution varies in different tissues and times in development. Unexpectedly, hypomorphic alleles of dcas show specific transformations in mechano-sensory organ cell identity, characteristic of mutations that increase Notch signaling. Dcas is essential for efficient importin alpha3 nuclear export in mechano-sensory cells and the surrounding epidermal cells and is indirectly required for the import of one component of the Notch pathway, but not others tested. We interpret the specificity of the dcas phenotype as indicating that one or more Notch signaling components are particularly sensitive to a disruption in nuclear protein import. We propose that mutations in house keeping genes often cause specific developmental phenotypes, such as those observed in many human genetic disorders. PMID- 11944948 TI - Divergent vascular mechanisms downstream of Sry establish the arterial system in the XY gonad. AB - Although the primitive vasculature is identical in XX and XY genital ridges until 11.5 days postcoitum (dpc), by 12.5 dpc the XY gonad develops a distinct vasculature. This male-specific vasculature, which includes the development of a large coelomic vessel, develops coincident with expression of Sry and formation of testis cords. We show that similar levels of proliferation and vasculogenesis expand the primary vasculature in XX and XY gonads. However, soon after Sry expression begins, the XY gonad recruits a large number of endothelial cells from the adjacent mesonephros, a mechanism totally absent in XX gonads. These migrating cells do not contribute to venous or lymphatic development. Instead, these cells contribute to the arterial system, as indicated by expression of ephrinB2 and by elements of the Notch signaling pathway. This newly formed arterial system establishes a new pattern of blood flow in the XY gonad, which we speculate may have an important role in export of testosterone to masculinize the XY embryo. PMID- 11944947 TI - The IGF pathway regulates head formation by inhibiting Wnt signaling in Xenopus. AB - The insulin-like growth factors (IGFs) are well known mitogens, both in vivo and in vitro, while functions in cellular differentiation have also been indicated. Here, we demonstrate a new role for the IGF pathway in regulating head formation in Xenopus embryos. Both IGF-1 and IGF-2, along with their receptor IGF-1R, are expressed early during embryogenesis, and the IGF-1R is present particularly in anterior and dorsal structures. Overexpression of IGF-1 leads to anterior expansion of head neural tissue as well as formation of ectopic eyes and cement gland, while IGF-1 receptor depletion using antisense morpholino oligonucleotides drastically reduces head structures. Furthermore, we demonstrate that IGF signaling exerts this effect by antagonizing the activity of the Wnt signal transduction pathway in the early embryo, at the level of beta-catenin. Thus, the IGF pathway is required for head formation during embryogenesis. PMID- 11944949 TI - Interaction of the essential Drosophila nuclear protein YA with P0/AP3 in the cytoplasm and in vitro: implications for developmental regulation of YA's subcellular location. AB - The Drosophila nuclear lamina protein YA is essential for the transition from female meiosis to embryo mitosis. Its localization and, hence, function is under developmental and cell cycle controls. YA protein is hyperphosphorylated and cytoplasmic in ovaries. Upon egg activation, YA is partially dephosphorylated and acquires the ability to enter nuclei. Its function is first detected at this time. To investigate the cytoplasmic retention machinery that keeps YA from entering nuclei, we used affinity chromatography and blot overlay assays to identify cytoplasmic proteins that associate with YA. Drosophila P0/AP3, a ribosomal protein that is also an apurinic/apyrimidinic endonuclease, binds to YA in ovary and embryo cytoplasms. P0 and YA bind specifically and directly in vitro and are present in a 20S complex in the cytoplasmic extracts. YA protein can be phosphorylated by MAPK, but not by p34(Cdc2) kinase, in vitro. This phosphorylation increases YA's binding to P0. We propose that the P0-containing 20S cytoplasmic complex retains hyperphosphorylated ovarian YA in the cytoplasm. In response to egg activation, YA is partially dephosphorylated and its binding to the 20S complex is reduced. Hence, some YA dissociates from the complex and enters nuclei. Consistent with this model, decreasing P0 levels partially suppress a hypomorphic Ya mutant allele. PMID- 11944950 TI - A retrospective study comparing the individual modalities of triple assessment in the pre-operative diagnosis of invasive lobular breast carcinoma. AB - AIMS: Early invasive lobular breast carcinoma (ILC) is associated with few symptoms and signs. The individual sensitivity of clinical examination, mammography, ultrasonography, cytology and core biopsy have each been reported to be of limited value. The aim of this study was to evaluate the accuracy of triple assessment in the pre-operative detection of patients identified to have ILC from their surgical pathology. METHODS: Pure ILC was defined as tumours containing at least 90% lobular features. The triple assessment of 273 patients diagnosed primarily at our institution were reviewed. RESULTS: 87.5% of women were symptomatic and 12.5% were screen detected. The mean patient age was 59 (range 30 81) years and the median tumour size was 26 (range 5-110) mm. The main mammographic abnormalities were a spiculated lesion (33.3%), an ill-defined mass (33.3%) or architectural distortion (23.5%). The sensitivities for detecting ILC of each modality were: clinical examination (76.6%), mammography (79.8%), ultrasound examination (93.9%), fine-needle aspiration cytology (FNAC) (60.5%) and core biopsy (90.8%). Combining the three modalities of clinical examination, imaging and cyto/pathology increased the pre-operative detection rate of ILC. CONCLUSION: Triple assessment is useful in the diagnosis of ILC. As the features of ILC may be subtle, a high index of suspicion is required to facilitate early diagnosis. PMID- 11944951 TI - Prognostic factors of gastric cancer tumours of less than 2 cm in diameter: rationale for limited surgery. AB - BACKGROUND: A recent trend in the surgical treatment of patients with early gastric cancer in Japan has been to limit surgery to an extent that ensures complete cure and improvement in the patient's quality of life. If a gastric cancer tumour can be completely eradicated by laparoscopic surgery, the patient can be cured of cancer without major operative stress. A small gastric cancer tumour of less than 2 cm in diameter is an indication for laparoscopic surgery, but little is known about what protocol of surgical treatment is appropriate for this type of tumour. PATIENTS AND METHODS: The clinicopathological features of 150 patients with gastric cancer tumour of less than 2 cm in diameter were reviewed retrospectively from hospital records between 1985 and 1995. The results of retrospective analysis of clinicopathological data of 24 patients with advanced cancer were compared with those of 126 patients with early cancer. Univariate and multivariate analyses of patients with small gastric cancer tumours were performed to evaluate the prognostic significance of clinicopathological features. RESULTS: A significant difference was seen between the gross tumour appearances in the two groups; Borrmann type-4 tumours were more common in the advanced group. Lymph-node metastasis, lymphatic vessel invasion and vascular invasion were found more frequently in the advanced cancer group than in the early cancer group. Scirrhous type was more common in the advanced cancer group. In univariate analysis, unfavourable prognostic factors included deep cancer invasion, presence of lymph-node metastasis, lymphatic invasion and vascular invasion. Using Cox's proportional hazard regression model, only nodal involvement emerged as an independent statistically significant prognostic parameter associated with long-term survival. CONCLUSION: Laparoscopic surgery should not be performed on tumours that are Borrmann type in macroscopic appearance and scirrhous-type histologically. Lymph-node metastasis is an independent prognostic factor. We recommend laparoscopic surgery involving local resection of the stomach without lymphadenectomy for small, early gastric cancer tumours that satisfy the criteria mentioned above. However, the validity of this recommendation should be tested by a prospective randomized control trial in the future. PMID- 11944952 TI - Ethnic survival differences after gastrectomy for gastric cancer are better explained by factors specific for disease location and individual patient comorbidity. AB - INTRODUCTION: Different outcomes after resection of gastric cancer between various ethnic patient groups have been described. It remains unclear whether disparity of treatment forms, disease-related variables, or individual patients accounts for this effect. METHODS: In the 10 years between 1989 and 1999, 75 patients with gastric adenocarcinoma underwent gastrectomy at a single institution, with constant surgical standards during this time period, including complete (R0) resection attempt and extended lymphadenectomy. Ethnicity, disease characteristics, and treatment variables were analysed for their impact on survival. RESULTS: There were 40 males and 35 females, with a median age of 67 years (range 31-97). The gastrectomy extent was total (n=25), proximal (n=18), subtotal (n=17), distal (n=14), and segmental (n=1). The mean lymph-node count was 25+/-17 (SD). There was one post-operative death, and an overall complication rate of 27%; the median hospital stay was 11 days. Overall actuarial 5-year survival was 33% (95% CI: 19-47); potentially curable disease (stage 1A-IIIB) led to a median survival of 49 months. Asian (n=18) and Hispanic patients (n=20) had significantly better survival than Caucasian (n=31) or other patients (n=6) (P=0.01). Ethnicity was linked to the location of the primary tumour ( P=0.002), the gastrectomy extent (P=0.003), and the patient's prior abdominal operation (P=0.01) or tobacco history (P=0.03), but not to resection extent parameters (such as number of lymph nodes retrieved) or differences in pathologic characteristics. When controlling for differences of disease site, stage, R status, and patient comorbidity, ethnicity did not retain an independent prognostic impact on survival. CONCLUSIONS: Obvious survival differences after gastrectomy for gastric adenocarcinoma favouring Asian and Hispanic patients in this experience can be explained by different disease patterns (distal location), the related need for fewer extensive procedures (such as total gastrectomy), and diminished patient risks (tobacco, prior operations, non-cancer deaths). Our therapeutic approach remains an aggressive gastrectomy/lymphadenectomy combination for potentially curable gastric cancer, irrespective of ethnic patient factors. PMID- 11944953 TI - Avoiding inappropriate surgery for secondary rectal cancer. AB - AIMS: Secondary rectal carcinoma occurs by invasion of the rectum by local primaries or by metastatic spread from a distant primary. The principle management of primary rectal carcinoma is surgery, but this is not usually the case for secondary carcinoma. This study investigates how these two may be differentiated and inappropriate surgery, in particular inappropriate abdominoperineal excision, can be avoided. METHODS: There were six patients with secondary carcinoma of the rectum. The primary tumours were: prostate (three), endometrium (two), breast (one). RESULTS: All the patients presented with lower gastrointestinal symptoms and four had a palpable mass on rectal examination. The diagnosis was made on histology and immunohistochemistry. Treatment was with endocrine therapy, chemotherapy, radiotherapy, and surgery. Three patients had palliative surgical procedures, and one had a curative anterior resection. The median survival was 7.5 months. CONCLUSIONS: This study has found that the presentation of primary and secondary rectal carcinoma is similar, and the method for distinguishing between the two is histology and immunohistochemistry. Staining for prostate-specific antigen was 100% accurate in the diagnosis of secondary rectal carcinoma arising from a prostate primary. The treatment of secondary rectal carcinoma is with systemic therapy and surgery is usually palliative, and therefore abdominoperineal excision should be avoided. PMID- 11944954 TI - Telomerase activity in human hepatocellular carcinoma: parallel correlation with human telomerase reverse transcriptase (hTERT) mRNA isoform expression but not with cell cycle modulators or c-Myc expression. AB - BACKGROUND: To explore the possible regulatory mechanisms of telomerase, we examined the telomerase activity (TA), expression of human telomerase RNA (hTR), human telomerase reverse transcriptase (hTERT) mRNA isoforms and cell cycle modulators in human hepatocellular carcinoma (HCC) cell lines (J5, J7) and a normal human immortalized hepatic epithelial cell line (Chang-liver). METHODS: The cell lines were chemically synchronized in either G1, G1/S, G2/M or M phases. TA was measured by polymerase chain reaction (PCR)-based telomerase repeat amplification protocol assay. The hTR and hTERT mRNA levels were analyzed by reverse transcriptase-polymerase chain reaction. Western blotting and immunocytochemistry were used to assay the cell cycle modulators. RESULTS: The TA of J5, J7 and Chang-liver cell lines tested was highest in M phase. The expression level of hTERT mRNA associated with the highest TA detected in the M phase of HCC cell lines. Chang-liver expressed markedly less TA and hTERT mRNA than J5 or J7. The elevated TA and expression of hTERT mRNA isoforms in M phase of HCC cell lines did not significantly correlate with that of the cell cycle modulators and c-Myc. CONCLUSIONS: The results implicate that regulation of TA is related to hTERT mRNA isoform expression, and that regulation is different between the cell immortalization and tumorigenesis. PMID- 11944955 TI - Computed tomographic features of hepatocellular carcinoma predict long-term survival after hepatic resection. AB - AIM: To evaluate the potential of computed tomographic features of hepatocellular carcinoma as prognostic factors. METHODS: Medical records for 112 patients who had undergone a partial hepatectomy for hepatocellular carcinoma were retrospectively analyzed. The largest hepatic tumour in each patient was classified by pre-operative computed tomographic features as lobular configurations with indentations showing an acute angle, or non-lobular configuration without such indentations. RESULTS: Twenty-six tumours were lobular and 86 were non-lobular. The outcome after hepatectomy was significantly worse in patients with lobular tumours (cumulative 5-year survival rate of 19.9%) than with non-lobular ones (that of 75.2%) (P<0.001). Cox's proportional hazards model showed computed tomographic features (P=0.0025), cirrhosis (P=0.0033), and tumour size (P=0.0412) to be independent prognostic factors. A lobular configuration was associated with satellite nodules (P<0.001), portal vein invasion (P=0.021), and extrahepatic tumour relapse (P=0.006). CONCLUSIONS: Computed tomographic features represent a strong prognostic factor in patients undergoing partial hepatectomy for hepatocellular carcinoma, and are likely to accurately reflect the tumour biology. Configuration, size of the hepatic tumour and presence of cirrhosis are the most important prognostic imaging findings in these patients. PMID- 11944956 TI - [131I]MIBG radionuclide therapy in carcinoid syndrome. PMID- 11944960 TI - Amphibian melanotrophs as a model to analyze the secretory plasticity of endocrine cells. AB - Precise regulation of hormone secretion from endocrine cells is of critical importance for the maintenance of animal homeostasis. This is partly enabled through the ability of endocrine cells to adapt dynamically their secretory activity to the physiological demands through complex functional changes. Such a secretory plasticity results from coordinated adaptive changes at several levels of cell function, including hormonal gene expression and biosynthesis, hormone processing, trafficking, storage and release, expression of membrane receptors, activation of signaling pathways, etc. Integration of all these processes at the single cell level defines the secretory status of each of the individual cells producing a given hormone, whose coordinated activity ultimately determines the response of the whole endocrine gland. This short review summarizes our most recent findings on the cellular and molecular elements and mechanisms underlying the secretory plasticity of endocrine cells, obtained from the analysis of distinct aspects of melanotroph cell function. PMID- 11944961 TI - Arginine vasotocin: site and mode of action in the reptilian kidney. AB - Recent morphological, physiological, and molecular studies that shed light on the functional significance of the neurohypophysial peptide arginine vasotocin (AVT) in the reptilian kidney are reviewed. Several structural features of the reptilian nephron are highlighted, including the presence of an incipient juxta glomerular apparatus, aglomerular nephrons, and the thin, very short intermediate segment (IS) joining proximal and distal convoluted tubules. Although the V(2) like AVT receptor and its gene have yet to be sequenced in any reptile, AVT receptors have been located in both the IS and the branched collecting duct system in the agamid lizard Ctenophorus ornatus. These findings suggest that AVT may have a dual effect in the reptilian kidney, first diluting the urinary fluid in the thin-intermediate segment, prior to its entering the collecting duct system, and then facilitating water reabsorption along an osmotic gradient as the urine passes through the final segments of the nephron. The IS may thus prove to be the evolutionary homologue of the thin-ascending limb of the loop of Henle in the mammalian kidney. PMID- 11944962 TI - Estrogen mimics bind with similar affinity and specificity to the hepatic estrogen receptor in Atlantic salmon (Salmo salar) and rainbow trout (Oncorhynchus mykiss). AB - A variety of organic chemicals have been documented to bind to and activate the estrogen receptor (ER) and consequently induce estrogenic effects in different animals. Although the function of the ER seems phylogenetically conserved in vertebrates, a considerable interspecies variation in the structure of the ER has been demonstrated. In this study, the hepatic ER from Atlantic salmon (Salmo salar) and rainbow trout (Oncorhunchus mykiss) were partly characterized, and the ligand-binding preference for a range of endogenous steroids and environmental estrogens (estrogen mimics) was determined by receptor-radio ligand studies. The results show that both Atlantic salmon and rainbow trout livers contain ERs that bind 1,2,4,6,7-[(3)H]estradiol ([(3)H]-E2) with high affinity and low capacity (K(d) = 2.5-4.4 nM and B(max) = 27-97 fmol/mg protein). The Atlantic salmon ER (asER) and rainbow trout ER (rtER) exhibit similar [(3)H]-E2 binding characteristics, although livers from female fish contained a two to three times higher amounts of ER than the males in the two species. In competition studies with [(3)H]-E2, the asER and rtER were found to bind both native steroids (E2 > estrone > 17 beta-estradiol 17-glucuronide >> testosterone and 11 ketotestosterone) and putative estrogen mimics (diethylstilbestrol, 4 hydroxytamoxifen, ethynylestradiol > genistein, zearalenone > 4-t-octylphenol, 4 n-nonylphenol, and o,p'-DDT). The pesticides toxaphen and dieldrin, which are proposed to bind to and activate the human ER, did not display significant binding affinity for the fish ER, however. In general, the asER and rtER were found to bind both native steroids and estrogen mimics with similar affinity and specificity. The present results suggest that closely related species such as Atlantic salmon and rainbow trout display similar ER ligand-binding requirements, although interspecies differences in ER affinity and specificity between divergent species such as fish and humans may exist. PMID- 11944963 TI - Testosterone triggers the brain-pituitary-gonad axis of juvenile female catfish (Heteropneustes fossilis Bloch) for precocious ovarian maturation. AB - The brain-pituitary-gonad axis of precociously matured females (PMFs) of Indian catfish (Heteropneustes fossilis), produced by testosterone treatment during juvenile stages, was analyzed by studies on immunoreactive gonadotropin-releasing hormone (ir-GnRH) secreting cells of the preoptic area of brain, plasma levels of gonadotropin (GtH-II), testosterone (T), and estradiol-17 beta (E(2)). GnRH cells of PMFs were large and strongly immunoreactive in comparison to control females. PMFs showed higher plasma levels of GtH-II, T, and E(2) than did control females. The ovaries of PMFs contained ripe ova, whereas control females had ova at maturing stages. This study suggests testosterone-mediated activation of the brain-pituitary-ovarian axis for precocious maturation in juvenile catfish. PMID- 11944964 TI - Development of neuropeptide Y-related peptides in the digestive organs during the larval stage of Japanese flounder, Paralichthys olivaceus. AB - cDNAs encoding three neuropeptide Y (NPY)-related peptides were identified from the Japanese flounder. One peptide corresponds to NPY, and its mRNA was expressed in the brain and retina. The second peptide corresponds to peptide YY (PYY), and its mRNA was expressed in the brain but surprisingly not in the digestive organs. This suggests that PYY lacks function as a digestive hormone in flounder. The third peptide corresponds to peptide Y (PY), and its mRNA occupied the brain and the endocrine cells of the intestine and the pancreatic accessory islets, but not of the principal islet. This suggests that PY has a dual role as a neuropeptide and a digestive hormone. In flounder larvae, PY cells appear in the intestinal epithelia at 3 days posthatching (dph), when larvae start feeding, in contrast to the endocrine pancreas at 30 dph. Therefore, the endocrine system of PY, which probably regulates the pancreatic enzyme secretion, develops synchronously with the start of feeding in the intestine but about 1 month later in the endocrine pancreas in flounder. PMID- 11944965 TI - Effects of dietary phytoestrogens in vivo and in vitro in rainbow trout and Siberian sturgeon: interests and limits of the in vitro studies of interspecies differences. AB - A study of the effects of dietary genistein on trout and sturgeon in vivo showed that sturgeon was sensitive to 20 ppm of genistein, whereas trout was not. To analyze the origin of this interspecies difference in sensitivity, a cell culture technique was developed with hepatocytes from sturgeon and compared to results obtained with hepatocytes from trout in the same system. The hepatocyte culture proved to be useful as bioassay for estrogenicity. Vitellogenin (VTG), assayed by a specific enzyme-linked immunosorbent assay, was used as a biomarker of the estrogenic activity. 17 beta-Estradiol, its glucuronide and sulfate derivatives, and estradiol analogues (ethynylestradiol and diethylstilbestrol) were tested. Nonestrogenic compounds such as androgens, progesterone, and cortisol were tested as negative controls. VTG production was monitored at doses ranging from 1 nM to 10 microM estradiol. Phytoestrogens, from the isoflavone family, were tested individually at increasing doses exhibiting dose response curves for concentrations from 500 nM to 10 microM. With tamoxifen, an antagonist of estrogen receptors, the estrogenic effect was partially reduced. The effect was the same with ICI182,780 in sturgeon, whereas the effect was the opposite in trout. The estrogenic potency of the isoflavones ranged differently between the two species in the following order: biochanin A < daidzein = formononetin < genistein < equol in trout and biochanin A < genistein < daidzein < formononetin < equol in sturgeon. Further, in sturgeon, formononetin was the most potent phytoestrogen in vitro, whereas its activity was weakest in vivo. These data suggest that one must reconsider the relevance of heterologous estrogenic tests and of homologous in vitro tests for estrogenic potency of chemicals. PMID- 11944966 TI - Comparison of R128Q mutations in human, ovine, and chicken leptins. AB - Human leptin and its R128Q mutant, as well as the R128Q mutants of ovine and chicken leptins, were prepared, expressed in Escherichia coli, refolded, and purified to homogeneity yielding electrophoretically pure, over 95% monomeric protein. R128Q mutations did not change the binding properties to BAF/3 cells stably transfected with the long form of human leptin receptor compared, respectively, to non-mutated human, ovine, and chicken leptins. In contrast, the biological activity tested in a proliferation assay in the same cells was drastically changed. Human leptin R128Q lost its activity and even became a weak antagonist, whereas the activities of ovine and chicken leptins were reduced 25- and 80-fold. If dimerization models were applicable leptin receptor activation, the present results would suggest that site 2 of the hormone was impaired. Two models, the human growth hormone:human growth hormone receptor (hGH:hGHR) (1:2) and the granulocyte-colony stimulating factor:granulocyte-colony stimulating factor receptor (GCSF:GCSFR) (2:2) complexes, were used for modeling. Superimposing the leptin structure on the hGH and GCSF models in the complex structures did not indicate any role for R128 in receptor binding. This made it impossible to correlate the results shown in the present work with the currently available models. Therefore, leptin may bind its receptors in a manner different than those proposed until now. PMID- 11944967 TI - Dynamic endocrine responses to stress: evidence for energetic constraints and status dependence in breeding male green turtles. AB - During reproduction, male vertebrates may exhibit a continuum of interactions between sex and adrenal steroids during stressful events, the outcome of which may be important in either reducing or promoting male reproductive success. We studied adult male green turtles (Chelonia mydas) to examine if they altered plasma corticosterone (CORT) and androgen levels in response to a standardized capture/restraint stressor as potential mechanisms to maintain reproductive activity during stressful events. At the population level, we found that migrant breeding males had a significantly smaller CORT response to the capture/restraint stressor compared to nonbreeding males and that this decreased response coincided with the generally poorer body condition of migrant breeders. In contrast, plasma androgen levels decreased significantly in response to the capture/restraint stressor in migrant breeding males, but not in nonbreeding and pre-migrant breeding males. For individual migrant breeding males, the magnitude of their CORT and androgen responses to the capture/restraint stressor was highly correlated with their body condition and body length, respectively. Our results demonstrate that male green turtles exhibit complex interactions in their endocrine responses to a capture/restraint stressor and that variation in these interactions is associated with differences in males' reproductive, energetic, and physical state. We hypothesize that interplay between physical status and plasma hormone responses to stressors could have important consequences for male green turtle reproduction. PMID- 11944968 TI - Effect of salt acclimatization on 3 beta-hydroxysteroid dehydrogenase/isomerase activity in the interrenal of Bufo arenarum. AB - In amphibians, aldosterone (Aldo) is particularly important in the regulation of Na(+) exchange by skin and urinary bladder. In previous works we studied a key enzyme in Aldo biosynthesis, the 3 beta-hydroxysteroid dehydrogenase/isomerase (3 beta HSD/I), in the interrenals of Bufo arenarum. In those works a dual localization of the 3 beta HSD/I in both microsomes and mitochondria was described. The mitochondrial, but not the microsomal, enzyme prefers the immediate Aldo precursor, 3 beta-analogue of aldosterone, as substrate. In this order, the enzyme 3 beta HSD/I would be not only a key enzyme for the synthesis of Aldo but additionally, due to its microsomal and mitochondrial localization, a possible target for the regulation of Aldo biosynthesis. With this rationale in mind, we have used in vivo and in vitro approaches to study Aldo regulation. In the present investigation the levels of Aldo were determined in plasma of winter (W) and summer (S) toads subjected to different saline concentrations (0.125 and 0.15 M) or kept on wet land. Saline hyperosmotically treated toads had significantly lower levels than isoosmotically treated toads. These results are consistent with the response in mammals, in which salt loading provokes a reduction in Aldo secretion. In W toads, plasmatic corticosterone (B) concentration was higher than Aldo concentration, whereas in S toads, B/Aldo ratio approached unity. The reduction of Aldo levels after saline dehydration was due to a decline in its biosynthesis. K(m) and V(max) values for 3 beta HSD/I were calculated for mitochondrial and microsomal fractions obtained from animals acclimated to 0.15 M NaCl or kept on land. As previously described, V(max) differs between W and S toads. However, only mitochondrial V(max) changed as a consequence of saline adaptation, suggesting that the mitochondrial enzyme could be involved in the regulation of Aldo biosynthesis. PMID- 11944969 TI - Production of recombinant goldfish prolactin and its applications in radioreceptor binding assay and radioimmunoassay. AB - Goldfish prolactin cDNA was subcloned into a pRSET A vector and expressed in Escherichia coli. Recombinant goldfish prolactin was expressed mainly as insoluble inclusion bodies in the form of N-terminal 6x His-tagged fusion protein. This fusion protein was purified, refolded, and (125)I-labeled to generate a radioligand for receptor binding and validation of a radioimmunoassay for goldfish prolactin. Using goldfish gill membrane as the substrate for prolactin receptor binding, both recombinant and native forms of goldfish prolactin were effective in displacing the specific binding of the radioligand in a similar dose range, suggesting that the fusion protein was refolded properly and could be recognized by goldfish prolactin receptors. To quantify prolactin contents in biological samples from the goldfish, a radioimmunoassay using the (125)I-labeled recombinant prolactin as a tracer was established. This assay was shown to be selective for goldfish prolactin without cross-reactivity with mammalian prolactin and pituitary hormones from other fish species (e.g., growth hormone and gonadotropin II). This newly validated assay system was used to investigate neuroendocrine and signal transduction mechanisms regulating prolactin release in the goldfish. In this case, the Ca(2+) ionophore A23187 and protein kinase C activator TPA were effective in elevating basal levels of prolactin secretion in perifused goldfish pituitary cells. In parallel studies using a static incubation approach, somatostatin and dopamine, but not vasoactive intestinal polypeptide, were inhibitory to basal prolactin release in goldfish pituitary cells. These results suggest that somatostatin and dopamine may serve as negative regulators of basal prolactin secretion and that extracellular Ca(2+) influx and protein kinase C activation may be important signaling events mediating prolactin release in the goldfish. PMID- 11944970 TI - Cold-stress induced the modulation of catecholamines, cortisol, immunoglobulin M, and leukocyte phagocytosis in tilapia. AB - The concentrations of norepinephrine in hypothalamus and norepinephrine and epinephrine in head kidney were significantly decreased in treated tilapia (Oreochromis aureus) during the time course of cold exposure (12 degrees) as compared to the control (25 degrees). The elevation of norepinephrine and epinephrine in plasma was detected earlier than that of cortisol in cold-treated tilapia. Phagocytic activity of leukocytes and the levels of plasma immunoglobulin M (IgM) were depressed in cold-treated tilapia as compared to the control group. Handling stress in the control (25 degrees) also resulted in increased plasma cortisol and decreased plasma IgM levels but not phagocytic activity. In vitro cortisol suppressed leukocyte phagocytosis in a dose (10(-12) to 10(-4) M)-dependent manner. Adrenergic agonist (phenylephrine and isoproterenol) had a significant suppression of phagocytosis only at the highest dose (10(-4) M). No effect on phagocytosis was detected in the treatment with norepinephrine and epinephrine. A combination of cortisol and isoproterenol (0.1 mM) had an additive effect in the suppression of phagocytosis. It is concluded that the cold stress modulated the changes of catecholamines and cortisol and further depressed phagocytic activity and antibody levels in tilapia. Cortisol could play a main and important role in the down-regulation of phagocytic activity. Adrenergic agonists also could interact with cortisol to further suppress immunity in tilapia. PMID- 11944971 TI - Effect of polar day on plasma profiles of melatonin, testosterone, and estradiol in high-Arctic Lapland Longspurs. AB - In polar habitats, continuous daylight (polar day) can prevail for many weeks or months around the summer solstice. In the laboratory, continuous light conditions impair or disrupt circadian rhythms in many animals. To determine whether circadian rhythms are disrupted under natural polar day conditions in a species that is only a summer resident in polar regions we analyzed diel rhythms in plasma concentrations of melatonin, testosterone (T), and 17-beta estradiol (E(2)) during the summer solstice in Arctic-breeding Lapland Longspurs (Calcarius lapponicus). We compared these profiles to those of conspecifics housed in outdoor aviaries at a mid-latitude site in Seattle, Washington, during spring, summer, fall, and winter. Under polar day conditions plasma melatonin concentrations of Lapland Longspurs were strongly suppressed, but still showed a significant diel rhythm. Likewise, plasma T in males, and E(2) in females, showed significant diel changes in Arctic birds. Lapland Longspurs housed at mid latitude in Seattle showed high-amplitude melatonin cycles at all times of the year, and the duration of the nightly melatonin secretion was positively correlated with the duration of the dark phase. We found no diel changes in plasma T in Seattle males in May, but Seattle females showed significant day/night differences in plasma E(2) in May. The data suggest that even under polar day conditions diel rhythms can persist. The maintenance of hormone rhythms could provide a physiological basis to reports of rhythmic behavior in many birds during the Arctic summer. PMID- 11944972 TI - Purification, characterization, and biological activity of insulins from the spotted dogfish, Scyliorhinus canicula, and the hammerhead shark, Sphyrna lewini. AB - Insulin was purified from pancreatic extracts of two elasmobranch species belonging to different families in the order Carcharhiniformes, the European spotted dogfish, Scyliorhinus canicula (Scyliorhinidae), and the hammerhead shark, Sphyrna lewini (Carcharhinidae). The amino acid sequence of dogfish insulin was established as A-chain GIVDHCCRNT(10)CSLYDLEGYC(20)NQ and B-chain LPSQHLCGSH(10)LVETLYFVCG(20)QKGFYYVPKV(30). The primary structure of hammerhead shark insulin was similar to that of dogfish insulin with only 2 amino acid substitutions at A8 (R --> H) and B30 (V --> I). The elasmobranch insulins were markedly different from human insulin (17 amino acid substitutions) but all the residues in human insulin that are believed to be important in determining the receptor binding conformation (B6, B8, B11, B13, B23, B24, B25, A2, A3, and A19) have been conserved in the elasmobranch insulins with the exception of the conservative substitution Phe --> Tyr at B25. Consistent with this, dogfish and human insulin showed almost identical binding affinity to the recombinant solubilized human insulin receptor (K(D) values of 14.0 and 18.6 pM, respectively; relative potency 133%). Previous studies have shown that bovine insulin produces severe and sustained hypoglycemia in elasmobranchs but the effect is of slow onset. Bolus arterial injections of dogfish insulin (10 nmol x kg(-1)) into unanesthetized, fasting dogfish (n = 9) produced no changes in blood glucose, 3-hydroxybutyrate, and acetoacetate concentrations over a 4-h period. In a second series of experiments (n = 7), dogfish insulin (10 nmol x kg(-1)) produced a significant (P < 0.05) fall in blood glucose after 12 h that persisted for at least 48 h, but no change in ketone body concentrations. The data indicate that the metabolic actions of an endogenous elasmobranch insulin in an elasmobranch are similar to those previously described for mammalian insulin. PMID- 11944974 TI - Guidelines for human gene nomenclature. PMID- 11944975 TI - Rules and guidelines for mouse gene, allele, and mutation nomenclature: a condensed version. PMID- 11944976 TI - Narrowing the critical region within 11q24-qter for hypoplastic left heart and identification of a candidate gene, JAM3, expressed during cardiogenesis. AB - Hypoplastic left heart is a severe human congenital heart defect characterized by left ventricular hypoplasiawith aortic and mitral valve atresia. A genetic etiology is indicated by an association of the hypoplastic left heart phenotype with terminal 11q deletions that span approximately 20 Mb (distal to FRA11B in 11q23). Here we define the breakpoints in four patients with heart defects in association with distal 11q monosomy and refine the critical region to an approximately 9-Mb region distal to D11S1351. Within this critical region we have identified JAM3, a member of the junction adhesion molecule family, as a strong candidate gene for the cardiac phenotype on the basis that it is expressed during human cardiogenesis in the structures principally affected in hypoplastic left heart. PMID- 11944977 TI - Transcriptional regulation of the human TATA binding protein gene. AB - The human TATA binding protein (TBP) locus consists of a functional domain of three closely linkedhousekeeping genes (TBP, PSMB1 (proteasomal C5 subunit), and PDCD2 (programmed cell death-2)) within a 50-kb interval at chromosome position 6q27. Here we demonstrate that a genomic clone spanning the 20-kb TBP gene, with 12 kb 5' and 3' flanking sequences, was fully functional in stable, transfected L cells harboring a single copy of this transgene, including after long-term (60 day) culture in the absence of drug selective pressure. Furthermore, we were only able to detect DNaseI hypersensitive sites at the TBP and PSMB1 promoters present within this 44-kb fragment. Our data suggest that this 44-kb genomic region possesses genetic regulatory elements that not only drive ubiquitous expression of TBP but also negate chromatin and DNA methylation induced silencing, which is normally associated with transgenes stably integrated into tissue culture cells. PMID- 11944978 TI - Imprinting of a small nucleolar RNA gene on mouse chromosome 12. AB - We have identified a novel, maternally expressed imprinted gene encoding a C/D box small nucleolar RNA (snoRNA) called MBII-343, which may regulate RNA editing or alternative splicing of an as yet unknown target gene. This gene is closely linked to an imprinted gene, Meg3, on mouse distal chromosome 12, which is syntenic to human chromosome 14. The paternal duplication of mouse distal chromosome 12 leads to late embryonal/neonatal lethality, growth promotion, and cardiomyopathy, whereas maternal duplication leads to late embryonal lethality and growth retardation. Human paternal uniparental disomy for chromosome 14 leads to musculoskeletal problems and mental retardation, whereas maternal uniparental disomy leads to intrauterine growth retardation, motor developmental delay, premature puberty, hypotonia, joint laxity, macrocephaly, short statue, neonatal poor sucking, skill with jigsaw puzzles, skin picking, obesity, and maturity onset diabetes of the young. PMID- 11944979 TI - Identification and characterization of a paralog of human cell cycle checkpoint gene HUS1. AB - A paralog of the human cell cycle checkpoint gene HUS1 has been identified and designated HUS1B. It encodes a 278-amino-acid protein, 48% identical and 69% similar to HUS1. Mouse and rat orthologs of HUS1B have also been detected by a BLAST search. HUS1B is expressed variably in many human tissues, and the tissue specific levels observed parallel those for HUS1. A HUS1-RAD1-RAD9 protein complex is thought to form a proliferating cell nuclear antigen (PCNA)-like structure, important for cell cycle checkpoint function. However, HUS1B directly interacts with RAD1, but not RAD9 or HUS1, whereas HUS1 can bind RAD1, RAD9, and another molecule of HUS1, suggesting that HUS1B cannot simply substitute for HUS1 in the complex. HUS1B is less conserved evolutionarily than HUS1. Furthermore, overexpression of HUS1B but not HUS1 in human cells induces clonogenic cell death. We suggest that HUS1B and HUS1 have distinct but related roles in regulating cell cycle checkpoints and genomic integrity. PMID- 11944980 TI - A t(2;8) balanced translocation with breakpoints near the human HOXD complex causes mesomelic dysplasia and vertebral defects. AB - Mesomelic dysplasia is a severe shortening of forearms and forelegs, and is found in several distinct human syndromes. Here, we report the cloning of the breakpoints of a human t(2;8)(q31;p21) balanced translocation associated with mesomelic dysplasia of the upper limbs, as well as with vertebral defects. We show that this translocation does not disrupt any gene, hence it most likely exerts its deleterious effect by modifying gene regulation. The HOXD complex lies approximately 60 kb from the translocation breakpoint on chromosome 2. This cluster of genes has an important role in the development of both the vertebral column and the limbs. Only a few cases of mutations of these homeotic genes have been described so far in humans. However, gain- and loss-of-function of Hoxd genes in mice can induce mesomelic dysplasia-like phenotypes, suggesting that misexpression of HOXD genes may indeed be at the origin of this hereditary phenotype. PMID- 11944981 TI - A new mouse model for autosomal recessive polycystic kidney disease. AB - In the course of large-scale mutagenesis studies, we discovered a mutant that provides a new mouse model for human autosomal recessive polycystic kidney disease. Animals homozygous for this mutation, T(2;10)67Gso, present evidence of grossly cystic renal and hepatic tissue at birth and a limited survival time of 3 4 days. The recessively expressed phenotype is associated with inheritance of a reciprocal translocation involving mouse chromosomes 2 and 10. Here we describe the pathology and phenotype of this new mutation. The mapping of the chromosomal breakpoint to the 1.0-cM critical region defined for another mouse autosomal recessive polycystic kidney disease model, juvenile congenital polycystic kidney disease (jcpk), led us to undertake the complementation testing that confirmed T(2;10)67Gso and jcpk are allelic. Because of the strong resemblance between the phenotype associated with these mouse mutations and early childhood polycystic kidney disease, and because of advantages offered by reciprocal translocations for gene mapping and cloning, T(2;10)67Gso should prove a valuable asset for studies concerning this fatal disease. PMID- 11944982 TI - Genetic control of polyamine-dependent susceptibility to skin tumorigenesis. AB - Overexpression of an ornithine decarboxylase (ODC) transgene greatly increases the susceptibility of mouse skin to carcinogen-induced tumor development. Like many phenotypes in transgenic models, this enhanced susceptibility phenotype is strongly influenced by genetic background. We have mapped tumor-modifier genes in intraspecific crosses between transgenic K6/ODC mice on a susceptible strain background (C57Bl/6J), a moderately resistant background (FVB), or a highly resistant background (C3H/HeJ). We identified several quantitative trait loci that influenced either tumor multiplicity or predisposition to the development of squamous cell carcinoma, but not both phenotypes. Because we did not use a tumor promotion protocol to induce tumors, most of the quantitative trait loci mapped in this study are distinct from skin tumor-susceptibility loci identified previously. The use of a combined transgenic-standard strain approach to genetic analysis has resulted in detection of previously unknown genetic loci affecting skin tumor susceptibility. PMID- 11944983 TI - Molecular cloning and characterization of the human S100A14 gene encoding a novel member of the S100 family. AB - S100 proteins form a growing subfamily of proteins related by Ca2+-binding motifs to the Efhand Ca2+-binding protein superfamily. By analyzing a human lung cancer cell line subtraction cDNA library, we have identified and characterized a new member of the human S100 family that we named S100A14 (GenBank acc. no. NM_020672). It encodes a mRNA present in several normal human tissues of epithelial origin, with the highest level of expression in colon. The full-length cDNA is 1067 nt in length, with a coding region predicting a protein of 104 amino acids that is 68% homologous to the S100A13 protein. The deduced amino acid sequence of the human S100A14 and its mouse homolog (identified as GenBank entry) contains two EF-hand Ca2+-binding domains, a myristoylation motif, a glycosylation site, and several potential protein kinase phosphorylation sites. We have mapped this gene to human chromosome 1q21, within a region where at least 15 other S100 genes are tightly clustered. A 3.2-kb genomic fragment containing the entire S100A14 was cloned and sequenced. The gene is split into four exons and three introns spanning a total of 2165 bp of genomic sequence. We examined the intracellular distribution of the epitope-tagged S100A14 protein in two human lung carcinoma cell lines and one immortalized monkey cell line. Pronounced staining was observed in the cytoplasm, suggesting an association with the plasma membrane and in the perinuclear area. We also provide evidence for heterogenic expression of S100A14 in tumors, demonstrating its overexpression in ovary, breast, and uterus tumors and underexpression in kidney, rectum, and colon tumors, a pattern suggesting distinct regulation with potentially important functions in malignant transformation. PMID- 11944984 TI - HBXAP, a novel PHD-finger protein, possesses transcription repression activity. AB - The PHD/LAP (plant homology domain/leukemia associated protein) finger motif is characteristically defined by a histidine and seven cysteines that are spatially arranged in a C4HC3 consensus sequence. This unique zinc finger, found primarily in a wide variety of chromatin-associated proteins, is considered to mediate protein-protein interactions. We have isolated a novel human PHD-finger protein, HBXAP (for hepatitis B virus x associated protein). HBXAP has three alternatively spliced isoforms. We also identified the Drosophila melanogaster HBXAP ortholog, gene CG8677. Based on alignment of four different proteins, we found a novel conserved domain in HBXAP that we designated the HBXAP conserved domain (XCD). We show that HBXAP represses transcription when recruited to DNA via the DNA binding of GAL4. Furthermore, the PHD finger alone suffices to repress transcription, thus attributing a functional role to this domain. The gene HBXAP is localized to the long arm of human chromosome 11 between q13.4 and q14.1. This region is amplified and rearranged in many tumors, suggesting a role for HBXAP in tumorigenesis similar to that of other PHD-containing proteins. PMID- 11944985 TI - Methylation dynamics of imprinted genes in mouse germ cells. AB - DNA methylation differences between maternal and paternal alleles of many imprinted genes are inherited from the male and female gametes and subsequently maintained during development. However, the stages of gametogenesis during which methylation imprints are established have not been well defined. In this study, we used bisulfite sequencing to determine the methylation dynamics of the imprinted genes small nuclear ribonucleoprotein N (Snrpn), insulin-like growth factor 2 receptor (Igf2r), mesoderm-specific transcript (Mest; formerly Peg1), paternally expressed gene 3 (Peg3), and H19 fetal liver mRNA (H19). We identified regions in the maternally imprinted genes (Snrpn, Mest, and Peg3) that were unmethylated in sperm but 100% methylated in mature oocytes. Igf2r, which is expressed from the maternal allele, was completely methylated within intronic differentially methylated region 2 in oocytes and unmethylated in sperm. The 5' region of H19, a paternally imprinted gene, was completely methylated in sperm and unmethylated in oocytes. We examined the methylation status of Snrpn during oocyte growth and maturation. Whereas the DNA of non-growing oocytes was mostly unmethylated, mid-size growing oocytes had a mosaic pattern of allelic methylation, and full acquisition of the methylation imprint was complete by metaphase II. We have identified regions within imprinted genes that show gamete specific methylation patterns in mature germ cells and demonstrated that maternal methylation imprints on at least one imprinted gene, Snrpn, are established during the postnatal growth phase of oogenesis. Thus, whereas paternal imprints seem to be established early (in diploid gonocytes well before the onset of meiosis), maternal imprints are established late (in growing oocytes that are arrested in the diplotene stage of meiosis). These findings raise the possibility that assisted reproductive technologies that involve in vitro maturation of oocytes may result in developmental abnormalities due to incomplete methylation imprints in immature oocytes. PMID- 11944986 TI - The apolipoprotein L gene cluster has emerged recently in evolution and is expressed in human vascular tissue. AB - We previously isolated APOL3 (CG12-1) cDNA and now describe the isolation of APOL1 and APOL2 cDNA from an activated endothelial cell cDNA library and show their endothelialspecific expression in human vascular tissue. APOL1-APOL4 are clustered on human chromosome 22q13.1, as a result of tandem gene duplication, and were detected only in primates (humans and African green monkeys) and not in dogs, pigs, or rodents, showing that this gene cluster has arisen recently in evolution. The specific tissue distribution and gene organization suggest that these genes have diverged rapidly after duplication. This has resulted in the emergence of an additional signal peptide encoding exon that ensures secretion of the plasma high-density lipoprotein-associated APOL1. Our results show that the APOL1-APOL4 cluster might contribute to the substantial differences in the lipid metabolism of humans and mice, as dictated by the variable expression of genes involved in this process. PMID- 11944987 TI - Magic roundabout is a new member of the roundabout receptor family that is endothelial specific and expressed at sites of active angiogenesis. AB - We have used bioinformatic data mining to identify a novel, endothelial-specific gene encoding a protein with homology to the axon guidance protein roundabout (ROBO1). The new gene has been called magic roundabout (ROBO4; GenBank acc. no. AF361473) and is smaller than other members of the roundabout gene family. Thus, in the extracellular region, magic roundabout has only two of the five immunoglobulin and two of the three fibronectin domains present in other roundabout genes. Expression of magic roundabout in vitro was detected in only endothelial cells and was greater in cells exposed to hypoxia. In situ hybridization and immunohistochemistry validated the bioinformatic prediction that magic roundabout expression would be endothelial specific in vivo. Magic roundabout expression in the adult was restricted exclusively to sites of active angiogenesis, notably tumor vessels. The identification of magic roundabout shows that the roundabout gene family extends beyond neuronal tissue and that roundabout/slit interactions are likely to have a role in angiogenesis. PMID- 11944988 TI - Increased gene dosage at Xq26-q27 is associated with X-linked hypopituitarism. AB - We have identified a novel interstitial duplication at Xq26.1-q27.3 in a previously reported family with X-linked recessive hypopituitarism [1]. Mapping of the duplication was carried out using interphase FISH analysis of over 60 bacterial genomic clones from Xq25-q28. The proximal and distal breakpoints of the duplication are contained within the 432N13 and 91O18 clones, respectively, and are separated by approximately 9 Mb. Comparison with a recently published 13 Mb duplication in another XH family [2] indicated that the duplication break points in these families were different. Therefore, we conclude that X-linked hypopituitarism is caused by increased dosage of a gene that is critical for pituitary development and that the causative gene is located within the 9-Mb duplicated region that we have defined. PMID- 11944989 TI - An integrated, functionally annotated gene map of the DXS8026-ELK1 interval on human Xp11.3-Xp11.23: potential hotspot for neurogenetic disorders. AB - Human chromosome Xp11.3-Xp11.23 encompasses the map location for a growing number of diseases with a genetic basis or genetic component. These include several eye disorders, syndromic and nonsyndromic forms of X-linked mental retardation (XLMR), X-linked neuromuscular diseases and susceptibility loci for schizophrenia, type 1 diabetes, and Graves' disease. We have constructed an approximately 2.7-Mb high-resolution physical map extending from DXS8026 to ELK1, corresponding to a genetic distance of approximately 5.5 cM. A combination of chromosome walking and sequence-tagged site (STS)-content mapping resulted in an integrated framework and transcript map, precisely positioning 10 polymorphic microsatellites (one of which is novel), 16 ESTs, and 12 known genes (RP2, PCTK1, UHX1, UBE1, RBM10, ZNF157, SYN1, ARAF1, TIMP1, PFC, ELK1, UXT). The composite map is currently anchored with 89 STSs to give an average resolution of approximately 1 STS every 30 kb. By a combination of EST database searches and in silico detection of UniGene clusters within genomic sequence generated from this template map, we have mapped several novel genes within this interval: a Na+/H+ exchanger (SLC9A7), at least two zincfinger transcription factors (KIAA0215 and Hs.68318), carbohydrate sulfotransferase-7 (CHST7), regucalcin (RGN), inactivation-escape-1 (INE1), the human ortholog of mouse neuronal protein 15.6, and four putative novel genes. Further genomic analysis enabled annotation of the sequence interval with 20 predicted pseudogenes and 21 UniGene clusters of unknown function. The combined PAC/BAC transcript map and YAC scaffold presented here clarifies previously conflicting data for markers and genes within the Xp11.3-Xp11.23 interval and provides a powerful integrated resource for functional characterization of this clonally unstable, yet gene-rich and clinically significant region of proximal Xp. PMID- 11944990 TI - Differential regulation of the human gene DAB2IP in normal and malignant prostatic epithelia: cloning and characterization. AB - Human DAB2IP (for DAB2 interaction protein) is a novel member of the RasGTPase activating protein family. It interacts directly with DAB2, which suppresses growth of many cancer types. We demonstrated that DAB2IP is often downregulated in human prostate cancer cell lines. The predicted DAB2IP protein (967 amino acids) shares 94.2% homology with the rat DIP1/2 protein. We mapped the promoter of DAB2IP and studied its regulation in normal and malignant prostate cancer cells. This gene is located at 9q33.1-q33.3 and spans approximately 96 kb with 15 exons and 14 introns. The DAB2IP promoter does not contain any typical TATA box evidenced by the presence of various RNAs with differential transcription starting sites. We further demonstrated that normal prostatic epithelial cells have elevated DAB2IP mRNA compared with cancer cells, which correlates with increased DAB2IP promoter activity. These data indicate that transcriptional regulation of DAB2IP is responsible for the downregulation of DAB2IP expression in prostate cancer cells. PMID- 11944991 TI - The identification of the inhibitory gamma-subunits of the type 6 retinal cyclic guanosine monophosphate phosphodiesterase in non-retinal tissues: differential processing of mRNA transcripts. AB - Here, we report that mouse lung expresses gamma-subunit (PDEgamma) transcripts of the rod and cone photoreceptor cGMP phosphodiesterase genes (Pde6g and Pde6h, respectively). Moreover, a major 14-kDa protein (p14) in lung membranes was immunostained with antibodies that react with both rod and cone PDEgamma. We show that p14 is, in fact, a mixture of rod and cone PDEgamma, based on three additional lines of evidence. First, p14 was also immunostained with antibodies specific for the cone PDEgamma isoform. Second, the expression of p14 immunostained with antibodies recognizing both rod and cone PDEgamma was substantially reduced in lung membranes from Pde6g-/- mice. In contrast, the fraction of p14 stained with cone PDEgamma-specific antibodies was not altered in the Pde6g-/- mice. Third, the absence of the Pde6g transcript was correlated with reduced levels of p14 in Pde6g-/- mice. We have also found that mouse lung contains a small Pde6h transcript that has a 41-bp deletion resulting in a frame change, derived by differential mRNA processing of exon 3 of Pde6h. BLAST searches also revealed a rat ovary EST that has the same 41-bp deletion causing the same frame change. However, the premature in-frame stop codon seen in the short Pde6h transcript is absent and the regular stop codon is out of frame leading to a predicted ORF extension into the 3' UTR. These findings show that rod and cone PDEgamma isoforms are expressed in lung and seem to have a critical role in regulating p42/p44 mitogen-activated protein kinase signaling. PMID- 11944992 TI - Analysis of the human neurexin genes: alternative splicing and the generation of protein diversity. AB - The neurexins are neuronal proteins that function as cell adhesion molecules during synaptogenesis and in intercellular signaling. Although mammalian genomes contain only three neurexin genes, thousands of neurexin isoforms may be expressed through the use of two alternative promoters and alternative splicing at up to five different positions in the pre-mRNA. To begin understanding how the expression of the neurexin genes is regulated, we have determined the complete nucleotide sequence of all three human neurexin genes: NRXN1, NRXN2, and NRXN3. Unexpectedly, two of these, NRXN1 ( approximately 1.1 Mb) and NRXN3 ( approximately 1.7 Mb), are among the largest known human genes. In addition, we have identified several conserved intronic sequence elements that may participate in the regulation of alternative splicing. The sequences of these genes provide insight into the mechanisms used to generate the diversity of neurexin protein isoforms and raise several interesting questions regarding the expression mechanism of large genes. PMID- 11944993 TI - Correct identification of genes from serial analysis of gene expression tag sequences. AB - SAGE (serial analysis of gene expression) is a remarkable technique for genome wide analysis of gene expression. It is crucial to understand the extent to which SAGE can accurately indicate a gene or expressed sequence tag (EST) with a single tag. We analyzed the effect of the size of SAGE tag on gene identification. Our observation indicates that SAGE tags are in general not long enough to achieve the degree of uniqueness of identification originally envisaged. Our observations also indicate that the limitation of using SAGE tag to identify a gene can be overcome by converting SAGE tags into longer 3' EST sequences with the generation of longer cDNA fragments from SAGE tages for gene identification (GLGI) method. PMID- 11944994 TI - Efficient cloning of full-length cDNAs based on cDNA size fractionation. AB - The ability to generate and obtain full-length (FL) cDNAs is of critical importance to the field of genomics. Most cDNAs in a traditional cDNA library lack the initiating 5' ATG, making it difficult to obtain a FL clone. We report here on an improved protocol for the preparation of FL enriched cDNA libraries. We demonstrate that if good quality RNA is used in the cDNA synthesis, high quality, FL cDNA can be generated for messages upward of 7 kb. In addition, we demonstrate the utility of size fractionation as a means to produce libraries containing a high percentage of initiating 5' ATG containing clones with insert sizes greater than 4 kb. The method is simple, cost efficient, and can be performed in most laboratories equipped to perform molecular biology. Lastly, the novel methodologies used in the analysis of the cDNA and library should prove useful to others working to create high-quality cDNA libraries. PMID- 11944995 TI - Genomic organization of the human gene HEP27: alternative promoter usage in HepG2 cells and monocyte-derived dendritic cells. AB - We used representational difference analysis to discover new genes with specific expression in dendritic cells. Among other genes, we identified HEP27, encoding a member of the short chain alcohol dehydrogenase/reductase family to be upregulated during monocyte to dendritic cell differentiation. Originally cloned from hepatocellular carcinoma cells (HepG2), HEP27 was exclusively expressed in monocyte-derived dendritic cells within the hematopoietic system. The presence of different transcripts in monocyte-derived dendritic cells, HepG2 cells, and various tissues could be traced back to alternative splicing and alternative promoter usage. We describe here the complete genomic organization of HEP27, including two alternative promoter regions: a hepatocyte-specific promoter which was induced by the histone deacetylase inhibitor sodium butyrate in several other cell types, and a second upstream promoter which was specifically active in monocyte-derived dendritic cells. Its exclusive usage in monocyte-derived dendritic cells makes the alternative HEP27 promoter an interesting target to study dendritic-cell-specific gene regulation. PMID- 11944996 TI - Immune regulation during pregnancy and host-pathogen interactions in infectious abortion. AB - The immunological mechanisms that govern the success of pregnancy in outbred mammals are complex. During placental formation the invasion of fetal cells into maternal tissue must be controlled to prevent damage to the mother. Equally, maternal recognition of pregnancy must be such that allorejection of the fetus does not occur. Despite the complexity of this phenomenon, it is clear that cytokines play a crucial role at the maternofetal interface and in the periphery to ensure that pregnancy proceeds successfully. Inflammatory cytokines such as tumour necrosis factor-alpha (TNF-alpha) and interferon-gamma (IFN-gamma) can exert detrimental effects in the placenta and tend to be present at low concentrations, whereas the regulatory cytokines interleukin (IL)-10 and tranforming growth factor-beta (TGF-beta) are beneficial and tend to predominate. This means that infection with pathogens that target the placenta and that elicit inflammatory responses may cause abortion by giving rise to a detrimental combination of cytokines that causes damage but does not control the disease. Infectious abortion is discussed in the context of the modulation of host immune responses during pregnancy, taking into account the different placental structures present in human beings, rodents and ruminants. PMID- 11944997 TI - Ground-glass hepatocytes in fibrinogen storage disease in Japanese Black calves. AB - This paper reports the occurrence of large intracytoplasmic inclusions observed in the hepatocytes of six Japanese Black calves showing clinical illness. These inclusions were round to elongated polyhedral in shape, with a consistently homogeneous glassy appearance. Hepatocytes with the inclusions had a ground-glass appearance. The inclusions were negative for the periodic acid-Schiff reaction and methenamine silver stain. Immunohistochemically, they were strongly positive for fibrinogen. Electron microscopy revealed that the inclusions consisted of granular material, showing moderate electron density and bounded by a unit membrane. On the external surface of the unit membrane, there were direct connections to cellular organelles, including the ribosomes and rough-surfaced endoplasmic reticulum. The results showed these inclusions to be entirely consistent with fibrinogen inclusions described in man. Hepatocellular fibrinogen storage disease, as identified in this study, has not previously been described in animals. PMID- 11944998 TI - Muscle lesions associated with dystrophin deficiency in neonatal golden retriever puppies. AB - Golden retriever muscular dystrophy (GRMD), a degenerative myopathy due to the absence of dystrophin, is genetically homologous to human Duchenne muscular dystrophy (DMD). Spontaneous death of GRMD neonates within the first 2 weeks of life occurs frequently. This report describes the microscopical muscle lesions that developed in 12 GRMD puppies aged 1-8 days of age, and makes a comparison with three normal age-matched siblings and two older GRMD dogs. Immunohistochemical methods were used to confirm dystrophin deficiency in GRMD puppies. Muscle lesions were assessed on sections stained with haematoxylin-eosin saffron, Gomori's trichrome and alizarin red S, and their severity was graded semi-quantitatively. Muscle fibre types were determined immunohistochemically on the basis of the pattern of expression of developmental, slow and fast isoforms of myosin. Muscle lesions in the GRMD puppies were characterized by massive necrosis, affecting most muscles of the proximal limbs, trunk and neck at birth. Lingual lesions began to develop in utero, and respiratory muscles underwent terminal diffuse necrosis resulting in death from acute respiratory failure. However, GRMD puppies do not invariably die in the neonatal period. Muscle in 2 month-old GRMD dogs showed signs of regeneration (immunohistochemical immaturity of muscle tissue), which suggested that all GRMD dogs suffer from massive post natal myonecrosis, whether fatal or not. Muscle lesions in neonates consisted mainly of hyalinization, hypertrophy, calcification and necrosis, followed by regeneration. Such "phase I" lesions due to the absence of dystrophin are found in all species in which dystrophin deficiency has been described (human beings, dogs, cats and mice), whereas the endomysial fibrosis and myofibre atrophy found in 2-month-old GRMD dogs constituted "phase II" lesions, which are specific to GRMD and human DMD. PMID- 11944999 TI - Immunohistochemical detection and distribution of inducible nitric oxide synthase in pigs naturally infected with Actinobacillus pleuropneumoniae. AB - Inducible nitric oxide synthase (iNOS) protein was detected immunohistochemically in formalin-fixed, paraffin wax-embedded lung tissues from 10 natural cases of porcine pleuropneumonia. Positive cells typically exhibited a red reaction product without background staining. Labelling of iNOS protein was intense in "oat cells", the clustered leucocytes with streaming nuclear chromatin that are a characteristic histological feature of porcine pleuropneumonia. Macrophages and neutrophils within alveolar spaces but not within blood vessels consistently showed iNOS labelling, but such labelling was minimal in non-lesional lung of Actinobacillus pleuropneumoniae -infected pigs and in normal lung from control pigs. The results suggest that iNOS plays a role in pathophysiological processes during A. pleuropneumoniae infection. PMID- 11945000 TI - Pathogenicity of Australian strains of avian infectious bronchitis virus. AB - The pathogenicity of 25 strains of infectious bronchitis virus (IBV) isolated in Australia between 1961 and 1994 was compared in white leghorn specific pathogen free chicks. Twelve strains were nephropathogenic and 10 respiratory, the other three being of mixed pathogenicity. The IBV strains identified as nephropathogenic induced clinical nephritis, gross and histological kidney lesions, and mortality of 5-90%. According to the severity of these features, the nephropathogenic strains could be further subdivided into strains of high, moderate or low pathogenicity. The three strains of mixed pathogenicity induced tracheitis, mild clinical nephritis and kidney lesions but no mortality. The 10 respiratory strains caused histological lesions in the trachea but not in the kidney, and did not induce clinical nephritis or mortality. Of 12 IBV strains isolated between 1961 and 1976, nine were nephropathogenic, inducing mortality of 15-90%. In contrast, of 13 strains isolated between 1981 and 1994, only three were nephropathogenic, inducing mortality of 5-37%, whereas nine were respiratory. Seven of these nine strains, unlike other respiratory strains, failed completely to replicate in the kidney. The results indicated a change in the prevalent IBV strains from highly nephropathogenic (1960s to 1970s) to respiratory (1980s to early 1990s); moreover, the late 1980s saw the emergence of respiratory strains with altered tissue tropism. PMID- 11945001 TI - Experimental Nipah virus infection in pigs and cats. AB - A human isolate of Nipah virus from an outbreak of febrile encephalitis in Malaysia that coincided with a field outbreak of disease in pigs was used to infect eight 6-week-old pigs orally or subcutaneously and two cats oronasally. In pigs, the virus induced a respiratory and neurological syndrome consistent with that observed in the Malaysian pigs. Not all the pigs showed clinical signs, but Nipah virus was recovered from the nose and oropharynx of both clinically and sub clinically infected animals. Natural infection of in-contact pigs, which was readily demonstrated, appeared to be acute and self-limiting. Subclinical infections occurred in both inoculated and in-contact pigs. Respiratory and neurological disease was also produced in the cats, with recovery of virus from urine as well as from the oropharynx. The clinical and pathological syndrome induced by Nipah virus in cats was comparable with that associated with Hendra virus infection in this species, except that in fatal infection with Nipah virus there was extensive inflammation of the respiratory epithelium, associated with the presence of viral antigen. Viral shedding via the nasopharynx, as observed in pigs and cats in the present study, was not a regular feature of earlier reports of experimental Hendra virus infection in cats and horses. The findings indicate the possibility of field transmission of Nipah virus between pigs via respiratory and oropharyngeal secretions. PMID- 11945002 TI - The pathogenicity of louping ill virus for mice and lambs. AB - Mice and lambs were infected with the LI/I, LI/31 or MA54 strain of louping ill virus (LIV) to provide information relevant to testing the efficacy and biosafety of a new generation of flavivirus vaccines based on a Semliki Forest virus (SFV) vector. Whereas clinical signs and neuropathological lesions were consistently severe in mice, the majority of lambs showed lesions of moderate severity and only lambs with severe lesions were clinically affected. For both species, dispersal of viral antigen occurred along neuronal cell processes, and neuronal degeneration and death were confirmed as central events after infection with LIV. In contrast to lambs, in which most lesions remained localized, mice showed widely dispersed lesions which were associated with less intense leucocytic infiltrates. Among the infiltrating cells, histiocytes predominated and apoptotic forms were prominent in severely affected animals. The intranasal route of infection provided an efficient avenue for entry of LIV into the brain and resulted in lesions which were more severe than those produced by subcutaneous or intraperitoneal inoculation. PMID- 11945003 TI - Chronic meningoencephalitis associated with Brucella sp. infection in live stranded striped dolphins (Stenella coeruleoalba). AB - A chronic, non-suppurative meningoencephalitis was found in three young striped dolphins (Stenella coeruleoalba) during routine neuropathological examination of marine mammals live-stranded on the Scottish coast. In all three dolphins the lesions were associated with the isolation of a Brucella sp. from the brain and with the immunohistochemical detection of brucella antigen. Moreover, antibodies to Brucella spp. were detected in the two dolphins that were subjected to serological examination. Immunohistochemical and serological examinations for morbillivirus antigen and antibodies, respectively, were negative in all cases. Although brucella infection of marine mammals has been extensively documented in recent years, its association with lesions and disease is less well recognized. The present report provides the first description of an association between Brucella sp. infection and neuropathological changes in a cetacean species. PMID- 11945004 TI - Systemic administration of histamine reduces reovirus type 2-induced insulitis in suckling DBA/1 mice. AB - Previously we suggested that reovirus type 2 (Reo-2) infection induced autoimmune insulitis, resulting in mild diabetes in suckling mice. The effect of histamine (a lymphocyte function suppressor) on Reo-2-induced insulitis was examined. Systemic histamine administration reduced the development of insulitis and blood glucose elevation. Endogenous interleukin-2 (IL-2) activity by splenic cells and the production of antibodies to pancreatic islet cells were reduced by histamine treatment. In addition, histamine treatment increased cyclic adenosine monophosphate (cAMP) concentrations in the plasma. These results further suggest that the insulitis seen in Reo-2 infection in suckling mice is induced by an immune reaction. PMID- 11945005 TI - Effect of beta-aminopropionitrile and hyaluronic acid on repair of collagenase induced injury of the rabbit Achilles tendon. AB - Collagenase was injected into the Achilles tendon of both hind legs of 10 clinically normal adult male New Zealand white rabbits. One month after induction of the injury, beta-aminoproprionitrile (BAPN) or hyaluronic acid (HA) was injected into the tendon core of the right hind leg of each rabbit, the left hind leg being left untreated. The treatment effects were evaluated by electron microscopy and analysis of the glycosaminoglycan (GAG) content of samples at 2 and 6 months post-treatment. At 2 months, collagen fibrils in tendons from both hind legs were relatively small in diameter, irregularly arranged, and interspersed with abundant active tenocytes as compared with those in normal tendon uninjured by collagenase. In the matrix, the amount of HA increased, but chondroitin-6-sulphate was eliminated. At 6 months, BAPN-treated tendons had small-diameter, regularly arranged collagen fibrils. HA-treated tendons, on the other hand, had large diameters, as well as regularly arranged collagen fibrils by comparison with non-treated tendon. The results suggest that HA, unlike BAPN, promoted healing. PMID- 11945007 TI - Pathological and immunohistochemical studies of choroid plexus carcinoma of the dog. AB - Choroid plexus carcinomas in four dogs (three male, one female) aged small middle 2.5 to 10 years, were examined by light microscopy and immunohistochemistry. The dogs showed progressive neurological signs including ataxia, seizures, vestibular disease and cranial nerve deficits, lasting for several months in some cases. Primary tumours were localized in the lateral (one case), third (one case), and fourth (two cases) ventricles. Hydrocephalus was evident at post-mortem examination in one case. In two cases the neoplastic cells closely resembled the structure of normal choroid plexus, with a distinct papillary pattern, composed of well-differentiated columnar epithelium. In the other two cases, cellular pleomorphism, nuclear atypia, increased mitotic activity and necrosis were observed. In all cases, dissemination of neoplastic cell clusters was detected within the subarachnoid space or the ventricular cavity. Immunohistochemical examination showed a multifocal labelling pattern for pankeratin and cytokeratin AE1 and diffuse vimentin positivity in poorly differentiated tumours. Well differentiated choroid plexus carcinomas showed multifocal immunoreactivity for cytokeratin AE3, multifocal to diffuse immunoreactivity for vimentin and occasional positivity for carcinoembryonic antigen. Epithelial membrane antigen, Ber EP4 and S-100 were negative in all cases. Glial fibrillary acidic protein labelling occurred only in a single, poorly differentiated tumour. Occasional reactions for proliferating cell nuclear antigen and MIB-1 were seen in two cases. It was concluded that at least two morphological and possibly phenotypic subtypes (well-differentiated and anaplastic) of choroid plexus carcinoma of the dog could be identified. PMID- 11945006 TI - Capsular polysaccharide conjugate vaccines against contagious bovine pleuropneumonia: Immune responses and protection in mice. AB - The immunogenicity of Mycoplasma mycoides subsp. mycoides small colony biotype (MmmSC) vaccines was investigated in BALB/c mice. Groups of mice were vaccinated with either (1) unconjugated capsular polysaccharide (CPS), (2) CPS covalently conjugated to ovalbumin via a carbodiimide reaction, (3) CPS non-covalently bound to latex microspheres, (4) CPS non-covalently complexed with rabbit anti-CPS IgG, and (5) whole inactivated, ultrasonically disrupted (WID) MmmSC. Only mice immunized with the CPS-ovalbumin conjugate exhibited a significant (P<0 small middle dot001) antibody response against CPS. Mice immunized with WID vaccine exhibited a high ELISA antibody titre against non-CPS (protein) antigens only. Mice given WID vaccine were immune against challenge with live MmmSC, and exhibited a significantly reduced degree of mycoplasmaemia (both in incidence and duration) as compared with non-vaccinated controls (P<0 small middle dot001). Mice immunized with the CPS-ovalbumin conjugate did not exhibit a reduction in mycoplasmaemia. The bactericidal activity of rabbit MmmSC-antiserum in an in vitro growth inhibition test was related to the CPS antibody titre. This was not observed with antisera from the vaccinated mice. None of the mouse antisera exhibited growth inhibiting activity, irrespective of a high CPS or protein antibody titre (CPS-ovalbumin or WID vaccine groups, respectively). Thus, it would seem that protection against an MmmSC-induced mycoplasmaemia in the mouse is based upon cell-mediated rather than humoral immunity. The results suggest that conjugation to ovalbumin significantly increases the antibody response to CPS in the mouse; the lack of bactericidal activity of mouse anti-CPS as compared with rabbit anti-CPS in vitro suggests either that the titre of growth inhibiting antibodies is lower in the mouse or that the mechanism of growth inhibition differs between antibodies of the two species. PMID- 11945008 TI - African swine fever: Expression of interleukin-1 alpha and tumour necrosis factor alpha by pulmonary intravascular macrophages. AB - To determine, in the acute form of African swine fever (ASF), the relationship between the appearance of pulmonary oedema and viral replication and expression of cytokines by pulmonary intravascular macrophages (PIMs), 14 pigs were inoculated intramuscularly with ASF virus (strain Espana'70) and killed in pairs on days 1-7 post-inoculation. Samples of lung were examined immunohistochemically and ultrastructurally. The immunohistochemical study was carried out with antibodies against interleukin-1 alpha (IL-1alpha), tumour necrosis factor-alpha (TNF-alpha), viral antigen of ASF (Vp73) and a myeloid marker (SWC3). Viral replication was observed mainly in PIMs, which at the same time showed intense activation, accompanied by the expression of IL-1alpha and TNF-alpha. The occurrence of interstitial oedema, neutrophil sequestration and fibrin microthrombi in septal capillaries coincided with high degrees of cytokine expression by infected PIMs. Alveolar macrophages did not show a significant change in cytokine expression as a result of ASF infection, and viral replication was detected in only a low percentage of these cells. PMID- 11945009 TI - Production of tumour necrosis factor-alpha (TNF-alpha) and reactive nitrogen intermediates by ovine peripheral blood leucocytes stimulated by Ehrlichia (Cytoecetes) phagocytophila. AB - Ehrlichia (Cytoecetes) phagocytophila, the causative agent of tick-borne fever in sheep and pasture fever in cattle, is an immunosuppressive, obligately intracellular rickettsia that invades granulocytes and monocytes of ruminants. Infected animals are known to suffer from a number of secondary infections. The mechanisms of immunosuppression are believed to be associated with physical or functional damage to leucocytes and the release of immunosuppressive substances. In the present study, the effects of E. phagocytophila on the production of tumour necrosis factor-alpha (TNF-alpha) and reactive nitrogen intermediates by ovine peripheral blood mononuclear cells (PBMCs) were investigated in vivo and in vitro. The concentration of TNF-alpha and nitrate in ovine sera were significantly increased during infection with E. phagocytophila, peak concentrations occurring at the peak period of rickettsiaemia. The addition of E. phagocytophila to cell cultures enhanced in-vitro production of TNF-alpha and nitric oxide by normal ovine PBMCs. PMID- 11945010 TI - Congenital spongiform change in the brain stem nuclei of a domestic kitten. AB - This report describes a novel spongiform change in the brain stem nuclei of a 9 month-old mixed breed kitten with neurological signs. Histologically, vacuoles were found in perineuronal spaces and neuropil, with mild to moderate astrocytosis in the brain stem nuclei. Vacuoles were not observed in the cytoplasm of neurons and no evidence of neuronal loss was found. Ultrastructurally, there were intramyelinic vacuoles with separation of lamellae at intraperiod lines and larger spaces formed by coalescence of ruptured vacuoles. Immunohistochemically, abnormal accumulation of prion protein (PrP) was not detected in the brain stem lesions. This is, to our knowledge, the first report of a feline spongiform change localized in the brain stem nuclei. PMID- 11945011 TI - Undifferentiated ganglioneuroblastoma in a sheep. AB - Ganglioneuroblastoma, presumably originating from the adrenal medulla, was found in an 18-month-old sheep. Histologically, the tumour was mainly composed of undifferentiated neuroblasts with neuropil and occasional ganglion cells. The morphological and immunohistochemical features with respect to intermediate filament proteins and the neuroendocrine nature of the tumour are described. The neoplastic cells were positive for neuron-specific enolase, synaptophysin and vimentin. PMID- 11945012 TI - Mutation of the ectodysplasin-A gene results in bone defects in mice. AB - Anhidrotic ectodermal dysplasia (EDA) is an X-linked, recessive genetic disease characterized by dysfunctional sweat glands, poorly developed teeth, and premature balding in human beings. This disorder results from mutations in the gene for ectodysplasin-A, a type II transmembrane protein with tumour necrosis factor-alpha domains. An animal model of EDA, the Tabby mouse, also has mutations in the ectodysplasin-A gene and defects similar to those of human beings with EDA. In addition to these defects, Tabby mice acquire deformities in the distal portion of their tails at 10-12 weeks of age. Whole-mount staining of the skeleton with Alizarin Red and Alcian Blue revealed that the tail defect resulted from vertebral fractures just distal to the epiphysis. Histological analysis demonstrated that the structure of both the epiphysis and the subepiphyseal zone of the tail vertebrae was dysplastic while the shaft of the diaphysis was relatively normal. The overall structure of the trabecular bone of these animals was examined through 3-dimensional microcomputed tomography of the tibia. This analysis indicated that Tabby mice had a mild increase in the interconnectivity of the intertwined trabecular bone network but that individual trabeculae were relatively normal. Since it has been determined recently that the ectodysplasin-A gene is expressed in the osteoblasts of developing human embryos, it appears likely that this gene plays a role in normal bone development. PMID- 11945013 TI - Spontaneous squamous cell carcinoma of the tongue and multiple bronchioloalveolar carcinomas in a Virginia opossum (Didelphis virginiana). AB - Two primary tumours, squamous cell carcinoma of the tongue and multiple bronchioloalveolar carcinomas, were diagnosed in a Virginia opossum (Didelphis virginiana). Two oral masses were located in the right ventrolateral surface of the tongue, near the frenulum, and the lungs contained multiple, widely distributed, nodular masses. Microscopically, the oral masses were composed of invasive cords of pleomorphic, polyhedral cells, typical of squamous cells. The multiple pulmonary masses consisted of non-ciliated, cuboidal, columnar, or occasionally polyhedral cells arranged in an alveolar pattern with multifocal areas of necrosis. This is the first report of spontaneous oropharyngeal squamous cell carcinoma in the Virginia opossum. However, multiple pulmonary adenomas have been reported previously in this species, the lesions being similar to those in sheep pulmonary adenomatosis (jaagsiekte). In the present study, immunohistochemical examination of the pulmonary tumours with a rabbit polyclonal antiserum to jaagsiekte retroviral capsid protein proved negative. PMID- 11945014 TI - Subacute liver necrosis after experimental infection with rabbit haemorrhagic disease virus (RHDV). AB - Rabbit haemorrhagic disease (RHD) is usually peracute to acute, while subacute to chronic disease is rare. This paper describes gross and histopathological findings in four out of 20 rabbits aged 14 weeks, experimentally infected with one of two German field isolates of RHD virus. Eight rabbits survived the infection for 10 days and were killed after four of them, infected with 100 to 10 000 haemagglutination units, had started to develop progressive jaundice. Histopathologically, icteric livers showed severe subacute centrilobular bridging necrosis with calcification, and proliferation of periportal hepatocytes and bile ducts. Positive-strand RHDV RNA was detected by in-situ hybridization, mainly in periportal macrophages. Loss of the normal hepatic architecture, reparation (fibrosis) and hepatocellular regeneration, together with moderate inflammatory reaction, are signs of liver cirrhosis. These signs, observed in young rabbits given small doses of RHD virus, are interpreted as an unusual outcome of experimental inoculation. PMID- 11945015 TI - Gastrinaemia, tissue gastrin concentration and G cell density in the antral mucosa of swine with and without gastric ulcer of the pars oesophagea. AB - Gastric ulcer of the pars oesophagea (GUPO) of the stomach, which occurs spontaneously in swine, is of unknown aetiology. Because gastrin is a potent secretagogue linked to acid secretion, this study was designed to investigate, in pigs with or without GUPO, (1) basal and food-stimulated serum gastrin concentrations, (2) the concentration of tissue gastrin, and (3) gastrin producing (G) cell density in the antral mucosa. Sixty-four pigs (32 with, and 32 without ulceration) were studied. Antral mucosa was obtained immediately after slaughter for assessment of G-cell number and tissue gastrin concentration. The presence or absence of GUPO was not significantly associated with either of the parameters measured. PMID- 11945017 TI - Taking the death toll after cardiomyocyte grafting: a reminder of the importance of quantitative biology. PMID- 11945016 TI - Generation of blood-derived dendritic cells in dogs with oral malignant melanoma. AB - Advances in treatment of human melanoma indicate that immunotherapy, particularly dendritic cell (DC) immunization, may prove useful. The aim of this study was to investigate whether blood-derived DCs could be generated from canine melanoma patients. Peripheral blood mononuclear cells were isolated from three such dogs and cultured with recombinant canine granulocyte-macrophage colony stimulating factor (GM-CSF), canine interleukin 4 and human Flt3-ligand for 7 days. The resulting cells demonstrated a typical dendritic morphology, and were enriched for cells expressing CD1a, CD11c and MHC II by flow cytometric analysis. Thus, canine blood-derived DCs can be generated in vitro and DC immunization should be feasible in dogs. PMID- 11945018 TI - Patched versus integrated analyses of the L-type Ca(2+) currents in aging myocytes. PMID- 11945019 TI - Calcineurin and the heartbeat, an evolving story. PMID- 11945020 TI - Late preconditioning elicited by activation of adenosine A(3) receptor in heart: role of NF- kappa B, iNOS and mitochondrial K(ATP) channel. AB - Activation of adenosine A(3) receptor (A(3)AR) protects against ischemia/reperfusion injury in the heart. However, the downstream signaling mechanisms leading to its delayed anti-ischemic effects remain unclear. We hypothesized that A(3)AR stimulation protects the heart via activation of nuclear transcription factor kappa B (NF-kappa B) and synthesis of inducible nitric oxide synthase (iNOS). Mice were treated with selective A(3)AR agonist, N(6)-(3 iodobenzyl) adenosine-5;-N-methyluronamide (IB-MECA). Twenty-four h later, hearts were perfused in Langendorff mode and subjected to 30 min of global ischemia and 30 min of reperfusion. IB-MECA caused post-ischemic reduction in necrosis and improvement in myocardial performance which was abolished by A(3)AR antagonist, MRS1191. Electrophoretic mobility shift assay demonstrated increased NF-kappa B binding in nuclear extracts following A(3)AR stimulation, which was diminished by MRS1191 and NF-kappa B inhibitor, pyrrolidinediethyldithiocarbamate (PDTC). The cardioprotection was abrogated by PDTC and targeted ablation of p50 subunit of NF kappa B in mice. The inhibition of iNOS with S-methylisothiourea and targeted disruption of the iNOS gene also abolished the protective effect of A(3)AR stimulation. Expression of iNOS mRNA and NO production were enhanced after 6 and 24 h respectively of IB-MECA treatment. MRS1191 and PDTC blocked IB-MECA induced NO production after A(3)AR stimulation. MitoK(ATP) channel blocker, 5 hydroxydecanoate abolished the protective effect of A(3)AR. For the first time, we have provided direct evidence of an essential role of NF- kappa B activation and iNOS in A(3)AR-induced late preconditioning. Selective activation of A(3)AR with IB-MECA can be used to trigger long-lasting ischemic protection in the heart. PMID- 11945021 TI - Blockade of T cell costimulatory signals using adenovirus vectors prevents both the induction and the progression of experimental autoimmune myocarditis. AB - Experimental autoimmune myocarditis (EAM) has been used as a model for human myocarditis in relation to the autoimmune mechanism and proved to be a T cell mediated autoimmune disease. Interactions of T cell surface receptors CD28 and CD40L with their ligands B7 and CD40, respectively, on APCs are critical for antigen-specific T cell activation under physiological and pathological conditions. To achieve effective inhibition of these interactions, we have constructed adenovirus vectors containing CTLA4Ig (AdexCTLA4Ig) and CD40Ig (AdexCD40Ig) and examined the effects of these adenovirus vectors in preventing EAM. AdexLacZ as a control, or AdexCTLA4Ig and/or AdexCD40Ig were injected intravenously into rats on day 0 or 14 after immunization to study the preventive effects on EAM in the T cell activation phase or inflammatory phase. Disease severity was estimated by the macroscopic and microscopic findings of the heart, heart weight to body weight ratios, and cellular and humoral immune responses on day 21. The onset of EAM after AdexCTLA4Ig or AdexCD40Ig treatment on day 0 was completely inhibited and antigen-specific lymphocyte proliferation was significantly reduced in those adenovirus-treatment groups, suggesting that those therapies induce antigen-specific T cell anergy. Moreover, significant reduction in disease severity was achieved after the adenovirus vector treatment even on day 14 compared with EAM rats. This study indicates the therapeutic potential of costimulatory pathway blockade by gene-transfer in myocarditis. PMID- 11945022 TI - Alterations in properties of L-type Ca channels in aging rat heart. AB - Previous studies of whole-cell L-type Ca currents in aging heart have demonstrated an increase in the peak Ca current magnitude in proportion to the increase in membrane area, and a slowing of the time course for inactivation. However, the single-channel mechanisms underlying this upregulation, and for the slowed inactivation are not known. We have therefore compared the properties of single L-type Ca channel currents recorded from ventricular myocytes obtained from young adult (3 month), adult (6-8 month) and aging (24 month) Wistar rats, using 5 m m Ba ions as the permeant ion. We report that the peak ensemble averaged single Ca channel currents from aging heart (-280+/-57 fA) were enhanced compared to those from young adult (-137+/-16 fA), or adult hearts (-144+/-38 fA). This surprising result was related, in part, to an apparent increase in the number of active Ca channels per patch in aging (1.90+/-0.23) v young adult (1.33+/-0.19) or adult heart (1.50+/-0.2). Moreover, there was an increase in the time constant for inactivation of the ensemble-averaged Ca currents of aging (471+/-169 ms), compared with young adult (198+/-43 ms), or adult heart (196+/-32 ms). The aging-related changes were also traced to alterations in single Ca channel gating, including an increase in the average probability of being open, and an increase in the availability of single Ca currents in aging heart. In contrast, the unitary Ca current amplitude was unchanged with aging. These novel findings suggest that the compensatory increase in the L-type Ca currents during aging is a consequence of an apparent increase in both the number, and the activity of individual L-type Ca channels. PMID- 11945023 TI - Ischemia-induced association of the stress protein alpha B-crystallin with I-band portion of cardiac titin. AB - During ischemia the cardiac stress protein, alpha B-crystallin, was shown by immunoelectron microscopy to translocate to the N(2)-line area of myofibrillar I bands of rat cardiomyocytes where alpha B-crystallin resisted extraction with 1 m NaSCN and 2 m urea as did titin. Actin became completely extracted under these conditions, indicating association of alpha B-crystallin with titin the only remaining non-actin cytoskeletal component of I-bands outside Z-disks. Titin, extracted from ischemic pig myocardium, was shown to copurify with alpha B crystallin. Further evidence for binding of alpha B-crystallin to titin was obtained by dot-blot assays in which biotinylated alpha B-crystallin was demonstrated to bind to the titin-enriched fraction immobilized on nitrocellulose. Binding of alpha B-crystallin to titin during cardiac ischemia could serve to stabilize titin against denaturation and might provide an endogenous mechanism to delay ischemic damage of this important elastic component of myofibrils. PMID- 11945024 TI - Evidence for calcineurin-mediated regulation of SERCA 2a activity in human myocardium. AB - Compromised SERCA 2a activity is a key malfunction leading to the Ca(2+) cycling alterations in failing human myocardium. SERCA 2a activity is regulated by the Ca(2+)/calmodulin-dependent protein kinase (CaM-kinase) but alterations of the CaM-kinase pathway regarding SERCA 2a in heart failure are unresolved. Therefore we investigated the CaM-kinase and phosphatase calcineurin mediated regulation of SERCA 2a in failing and non-failing human myocardium. We studied human myocardial preparations from explanted hearts from non-failing organ donors (NF, n=8) and from patients with terminal heart failure undergoing cardiac transplantation (dilated cardiomyopathy, DCM, n=8). SERCA 2a activity was determined using a NADH coupled enzyme assay [expressed in nmol ATP/(mg protein x min)] and by(45)Ca(2+) uptake. Protein expression of SERCA 2a, phospholamban, calsequestrin and calcineurin was assessed by Western blotting (expressed as densitometric units/microg protein); phosphorylation of cardiac proteins was detected with specific phospho-antibodies for phospholamban at threonine-17 (PT17) or by incorporation of [gamma -(32)P] (expressed as pmol(32)P/mg). Maximal(45)Ca(2+) uptake (in pmol/mg/min) (NF: 3402+/-174; DCM: 2488+/-189) and maximal SERCA 2a activity were reduced in DCM compared to NF (V(max): NF: 125+/-9; DCM: 98+/-5). The V(max) reduction could be mimicked by calcineurin in vitro in NF (NF(control): 72.1+/-3.7; NF(+calcineurin): 49.8+/-2.9) and restored in DCM by CaM-kinase in vitro (DCM(control): 98+/-5; DCM(+CaM-kinase): 120+/-6). Protein expression of SERCA 2a, phospholamban and calsequestrin remained similar, but calcineurin expression was significantly increased in failing human hearts (NF: 11.6+/-1.5 v DCM: 17.1+/-1.6). Although the capacity of endogenous CaM-kinase to phosphorylate PT17 was significantly higher in DCM (DCM(control): 128+/-36; DCM(+endogenous CaM-kinase): 205+/-20) compared to NF myocardium (NF(control): 273+/-37; NF(+endogenous CaM-kinase): 254+/-31), net phosphorylation at threonine 17 phospholamban was significantly lower in DCM (DCM 130+/-11 v NF 170+/-11). A calcineurin-dependent dephosphorylation of phospholamban could be mimicked in vitro by incubation of NF preparations with calcineurin (NF(control) 80.7+/-4.4 v NF(+calcineurin) 30.7+/-4.1, P<0.05). In human myocardium, the V(max) of SERCA 2a and the phosphorylation of phospholamban is modulated by CaM-kinase and calcineurin, at least in vitro. In failing human myocardium, despite increased CaM-kinase activity, calcineurin dephosphorylation leads to decreased net phosphorylation of threonine-17 phospholamban in vivo. Increased calcineurin activity contributes to the impaired V(max) of SERCA 2a in failing human myocardium and the disorder in Ca(2+)-handling in heart failure. PMID- 11945025 TI - Hypoxic preconditioning triggers myocardial angiogenesis: a novel approach to enhance contractile functional reserve in rat with myocardial infarction. AB - A modern experimental strategy for treating myocardial ischemia is to induce neovascularization of the heart by the use of "angiogens", mediators that induce the formation of blood vessels, or angiogenesis. Studies demonstrated that coronary collateral vessels protect ischemic myocardium after coronary obstruction; therefore we sought to examine a novel method of stimulating myocardial angiogenesis through hypoxic preconditioning at both capillary (using anti-CD31) and arteriolar (using anti- alpha smooth muscle actin) levels and also investigate whether such treatments could preserve left ventricular contractile functional reserve and regional blood flow by increasing vascular endothelial growth factor (VEGF). Male Sprague-Dawley rats were randomly divided into four groups: normoxia+sham surgery (CS), normoxia+permanent left anterior descending coronary artery (LAD) occlusion (CMI), hypoxic preconditioning+sham surgery (HS) and hypoxic preconditioning+permanent LAD occlusion (HMI). Rats in the preconditioned groups were subjected to systemic hypoxemic hypoxic exposure (10+/ 0.4% O(2)) for 4 h followed by a 24 h period of normoxic reoxygenation prior to undergoing LAD occlusion. Rats in the normoxia group were time matched with the preconditioned group and maintained under normoxic conditions for a 28 h period prior to LAD occlusion. Western blot analysis was performed to measure VEGF expression and TUNEL staining with endothelial cell-specific antibody, anti-VWF, was used to examine endothelial apoptosis. One, two and three weeks after the LAD occlusion, baseline left ventricular pressures were monitored and recorded. Pharmacological stress tests with dobutamine infusion in progressively increasing doses revealed significantly elevated contractile reserve at each dose point in the HMI group compared to the CMI group. The HMI group displayed statistically significant increases in capillary as well as arteriolar density after 1, 2 and 3 weeks post-operation. Blood flow was also significantly elevated in the HMI groups when compared to the CMI group. The extent of endothelial cell apoptosis was found to be inversely proportional to VEGF expression. It was concluded that hypoxic preconditioning stimulates myocardial angiogenesis to an extent sufficient to exert significant cardioprotection in a rat model of myocardial infarction progressing to heart failure as evidenced by increased capillary/arteriolar density and enhanced ventricular contractile functional reserve. PMID- 11945026 TI - Endothelial IKK beta signaling is required for monocyte adhesion under laminar flow conditions. AB - Endothelial activation induces expression of pro-inflammatory molecules that are thought to play an important role in atherogenesis through enhanced vascular monocyte recruitment. Many pro-inflammatory endothelial signals are transcriptionally regulated by members of the NF- kappa B family. The serine threonine kinase, IKK beta, can mediate NF- kappa B activation although several alternative pathways exist. To test whether IKK beta is necessary for cytokine activation of human vascular endothelium and endothelial recruitment of human monocytes under laminar flow, we constructed a recombinant adenoviral vector carrying a dominant negative mutant of IKK beta (Ad.dnIKK beta) to transduce human umbilical vein endothelial cells (HUVEC) in vitro. We found that dnIKK beta expression effectively blocked NF-kappa B activation as assessed by nuclear translocation of NF-kappa B, I kappa B degradation, and NF-kappa B dependent reporter expression, without affecting activation of the other relevant signaling pathways, SAPK/JNK and p38. Furthermore, overexpression of dnIKK beta in TNF alpha-stimulated HUVEC blocked induction of the surface adhesion molecules E selectin, ICAM-1, and VCAM-1. Under simulated physiologic flow conditions, both firm adhesion and rolling of human peripheral monocytes on dnIKK beta-transduced endothelial monolayers were markedly inhibited. We conclude that IKK beta is necessary for the cytokine-induced inflammatory phenotype of human endothelium and endothelial recruitment of human monocytes under flow. PMID- 11945027 TI - Expression of activated PKC epsilon (PKC epsilon) protects the ischemic heart, without attenuating ischemic H(+) production. AB - PKC epsilon is a PKC isoform that translocates during preconditioning and may mediate cardioprotection. To investigate whether PKC epsilon activation is cardioprotective, Langendorff-perfused hearts from wild-type (WT) mice and from mice expressing constitutively active mutant PKC epsilon were subjected to 20 min ischemia and 40 min reperfusion while(31)P NMR spectra were acquired. Pre ischemic glycogen levels were similar in WT and PKC epsilon hearts. During ischemia, ATP fell less in PKC epsilon than in WT hearts. Ischemic intracellular pH, however, was similar in WT and PKC epsilon hearts. During reperfusion, recovery of contractile function and ATP were greater in PKC epsilon than WT hearts. In conclusion, expression of activated PKC epsilon protected hearts from post-ischemic energetic and contractile dysfunction, consistent with the proposed cardioprotective role of PKC epsilon. Protection occurred in the PKC epsilon hearts without attenuation of ischemic H(+) production, implying that, at least in this ischemic model, reduced acidification during ischemia is not necessary for cardioprotection. PMID- 11945028 TI - A 3-D structural model of solid self-assembled chlorophyll a/H(2)O from multispin labeling and MAS NMR 2-D dipolar correlation spectroscopy in high magnetic field. AB - Magic angle spinning (MAS) NMR with Lee-Goldburg cross-polarization (LG-CP) is used to promote long-range heteronuclear transfer of magnetization and to constrain a structural model for uniformly labeled chlorophyll a/H(2)O. An effective maximum transfer range d(max) can be determined experimentally from the detection of a gradually decreasing series of intramolecular correlations with the (13)C along the molecular skeleton. To probe intermolecular contacts, d(max) can be set to approximately 4.2 A by choosing an LG-CP contact time of 2 ms. Long range (1)H-(13)C correlations are used in conjunction with carbon and proton aggregation shifts to establish the stacking of the chlorophyll a (Chl a) molecules. First, high-field (14.1 T) 2-D MAS NMR homonuclear ((13)C-(13)C) dipolar correlation spectra provide a complete assignment of the carbon chemical shifts. Second, proton chemical shifts are obtained from (1)H-(13)C heteronuclear dipolar correlation spectroscopy in high magnetic field. The shift constraints and long-range (1)H-(13)C intermolecular correlations reveal a 2-D stacking homologous to the molecular arrangement in crystalline solid ethyl-chlorophyllide a. A doubling of a small subset of the carbon resonances, in the 7-methyl region of the molecule, provides evidence for two marginally different well-defined molecular environments. Evidence is found for the presence of neutral structural water molecules forming a hydrogen-bonded network to stabilize Chl a sheets. In line with the microcrystalline order observed for the rings, the long T(1)'s, and absence of conformational shifts for the (13)C in the phytyl tails, it is proposed that the Chl a form a rigid 3-D space-filling structure. Probably the only way this can be realized with the sheets is by forming bilayers with interpenetration of elongated tails. Such a 3-D space-filling organization of the aggregated Chl a from MAS NMR would match existing models inferred from electron microscopy and low-resolution X-ray powder diffraction, while a micellar model based on neutron diffraction and antiparallel stacking observed in solution can be discarded. PMID- 11945029 TI - A robust technique for two-dimensional separation of undistorted chemical-shift anisotropy powder patterns in magic-angle-spinning NMR. AB - A robust magic-angle-spinning experiment for separating undistorted, quasi-static chemical-shift powder patterns is presented. It is derived from the technique of R. Tycko, G. Dabbagh, and P. Mirau (1989, J. Magn. Reson. 85, 265), but uses 360 degrees instead of 180 degrees pulses. In combination with a suitable phase sequence, this makes the spectral patterns very insensitive to pulse-length errors and other imperfections, as demonstrated both experimentally and theoretically. This method, termed separation of undistorted powder patterns by effortless recoupling (SUPER), can be used at standard spinning speeds, between 2.5 and 5 kHz, and with standard radiofrequency power levels (in particular, for protons, a decoupling field strength gammaB(1)/2pi of less than 80 kHz). No significant artifacts are observed even for samples extending to the ends of the radiofrequency coil. The method has been applied to samples containing various sp(2)- and sp(3)-hybridized carbon sites. Even for the methylene groups in polyethylene, a system with very strong C-H and H-H dipolar couplings and only moderate chemical-shift anisotropy (CSA), a useful CSA powder pattern has been obtained. With a CSA scaling factor of 0.155, accuracies of +/-5, +/-3, and +/-1 ppm of the principal values can be achieved for protonated aromatic carbons, unprotonated sp(2)-hybridized groups, and aliphatic sites, respectively. Examples of CSA-based assignment of COOC vs other COO or CON groups, and of aromatic vs olefinic C=C carbons are shown, for both small molecules and polymers. PMID- 11945030 TI - Cross-correlation effects involving curie spin relaxation in methyl groups. AB - Cross-correlation effects arising in methyl protons due to the simultaneous presence of dipole-dipole, chemical shift anisotropy, and Curie spin relaxation mechanisms in paramagnetic systems are analyzed. We assess the potential of obtaining structural constraints from the cross-correlation of Curie spin relaxation with dipolar relaxation mechanisms among methyl proton spins. By theoretical analysis and numerical simulations we characterize the transfer functions describing the interconversion processes of different ranks of multispin order. The time dependence of these processes contains a new type of structural information, the orientation of the methyl C(3)-axis with respect to the electron center. Experimental confirmation is found for selected methyl groups in low spin Fe(3+) sperm whale myoglobin. PMID- 11945032 TI - Adsorption and desorption behavior of NO on H-ZSM-5, Na-ZSM-5, and Na-A as studied by EPR. AB - Nitric monoxide probe molecules are used to characterize the Lewis acid properties of sodium cations and aluminum defect centers in various zeolite materials. The adsorption-desorption behavior of NO probe molecules is studied at different temperatures for Na-A, Na-ZSM-5, H-ZSM-5, and silicalite. Adsorbed NO molecules form paramagnetic adsorption complexes with Lewis acid sites which can be examined by EPR transitions ((Delta)m(S)+/-1) at g approximately 2.0. Otherwise the desorption of NO into the gas phase can be monitored by the typical nine-line EPR spectrum ((Delta)m(J)+/-1) of the (2)Pi(3/2) state at g approximately 0.7776. This gas-phase signal is used to study the overall adsorption-desorption properties of the zeolites in the temperature range 150 K less than or approximately T less than or approximately 300 K. At lower temperatures the probe molecules are adsorbed at the Lewis acid sites inside the nanoporous materials and produce an intensive spectrum at T less than or approximately 110 K. But at intermediate temperatures 110 K less than or approximately T less than or approximately 150 K the NO molecules are adsorbed only for a few hundred picoseconds because the lifetime of the adsorption complexes is limited by the beginning desorption processes. The decreasing lifetime of the adsorption complex with rising temperature results in an increasing homogeneous line broadening of their EPR signals. An analysis of the line-broadening effects provides an opportunity for determining the specific desorption energies E(A)(H-ZSM-5)=(20.2+/-7.3) kJ/mol, E(A)(Na-ZSM-5)=(4.1+/-1.5) kJ/mol, and E(A)(Na-A)=(7.1+/-2.1) kJ/mol for NO probe molecules at sodium cations and aluminum defect centers just below the desorption temperature. PMID- 11945031 TI - Automatic tuned MRI RF coil for multinuclear imaging of small animals at 3T. AB - We have developed an MRI RF coil whose tuning can be adjusted automatically between 120 and 128 MHz for sequential spectroscopic imaging of hydrogen and fluorine nuclei at field strength 3 T. Variable capacitance (varactor) diodes were placed on each rung of an eight-leg low-pass birdcage coil to change the tuning frequency of the coil. The diode junction capacitance can be controlled by the amount of applied reverse bias voltage. Impedance matching was also done automatically by another pair of varactor diodes to obtain the maximum SNR at each frequency. The same bias voltage was applied to the tuning varactors on all rungs to avoid perturbations in the coil. A network analyzer was used to monitor matching and tuning of the coil. A Pentium PC controlled the analyzer through the GPIB bus. A code written in LABVIEW was used to communicate with the network analyzer and adjust the bias voltages of the varactors via D/A converters. Serially programmed D/A converter devices were used to apply the bias voltages to the varactors. Isolation amplifiers were used together with RF choke inductors to provide isolation between the RF coil and the DC bias lines. We acquired proton and fluorine images sequentially from a multicompartment phantom using the designed coil. Good matching and tuning were obtained at both resonance frequencies. The tuning and matching of the coil were changed from one resonance frequency to the other within 60 s. PMID- 11945033 TI - Low-symmetry spin hamiltonian and crystal field tensors analysis: Fe(3+) in natrolite. AB - Electron paramagnetic resonance study of a natural single crystal of natrolite was carried out at the frequency nu=36.772 GHz at room temperature. The angular dependence of the four symmetry-related spectra of Fe(3+) in the three crystallographic planes was fitted to a spin Hamiltonian (S=5/2) of symmetry C(i). The rank 4 crystal field tensors at tetrahedral sites were calculated using the point-charge model to determine the principal axes orientations of their cubic and trigonal components. The analysis of zero-field splitting tensors and comparison with crystal field ones suggests that Fe(3+) substitutes for Al(3+) with no significant distortion of the coordination tetrahedron in natrolite. Comparison of data for several natural and synthetic crystals reveals that the 4 rank zero-field splitting tensor invariants for Fe(3+) at the tetrahedral oxygen coordinated sites are distinguishably smaller than those for Fe(3+) at octahedral sites. Such comparative analysis may help to determine the substitutional sites in other crystals. PMID- 11945034 TI - 1H detected (13)C echo planar imaging. AB - Cyclic J cross polarisation (CYCLCROP) is a sensitive method for the noninvasive monitoring of (13)C distributions and fluxes. The PRAWN rotating frame Hartmann Hahn mixing sequence ameliorates problems associated with sensitivity to Hartmann Hahn mismatch and reduces RF power deposition. The combination of CYCLCROP with echo planar imaging (EPI) for spatial encoding of the proton detected carbon signal allows efficient use of the available signal to be made, permitting a significant improvement in the temporal resolution of any study. We report here on some initial experiments to demonstrate the feasibility of echo planar proton detected (13)C imaging using CYCLCROP based upon the PRAWN module, including the application of the technique to the measurement of transport and accumulation of (13)C-labelled sucrose in a castor bean seedling. Two methods that can be used to eliminate the effect of the J-splitting in the EP images are presented. In addition, a fast, image-based B(1) field-mapping method which may be used to quantitatively map the low frequency RF field in a dual resonant ((13)C/(1)H) probe is presented. The technique utilises the above described imaging method, permitting fully quantitative, 64x64 axial field maps to be generated in about a minute. PMID- 11945035 TI - Direct conversion of EPR dipolar time evolution data to distance distributions. AB - Shallow electron spin echo envelope modulations due to dipole-dipole couplings between electron spins provide information on the radial distribution function of the spins in disordered systems while angular correlations between spin pairs are negligible. Under these conditions and in the absence of orientational selection, the dipolar time evolution data can be quantitatively simulated for arbitrary radial distribution functions by shell factorization, i.e., by performing the orientational average separately for thin spherical shells and multiplying the signals of all the shells. For distances below 5 nm, a linear superposition of the signals of the shells is sufficient. The dipolar time evolution data can be separated into this linear contribution and a nonlinear background. The linear contribution can then be converted directly to a radial distribution function. For a series of shape-persistent and flexible biradicals with end-to-end distances between 2 and 5 nm, shell factorization and direct conversion of the data are in good agreement with each other and with force-field computations of the end-to-end distances. The neglect of orientation selection does not cause significant distortions of the determined distance distributions. PMID- 11945036 TI - Efficiency of homonuclear Hartmann-Hahn and COSY-type mixing sequences in the presence of scalar and residual dipolar couplings. AB - In the presence of scalar (J) and residual dipolar (D) couplings, the transfer efficiency of homonuclear Hartmann-Hahn and COSY-type mixing depends on the ratio D/J and on the mixing sequence. This dependence is analyzed theoretically and the results are confirmed experimentally. At least two different mixing sequences are required to yield good transfer efficiencies for all ratios D/J. In contrast to COSY-type experiments, homonuclear Hartmann-Hahn sequences can provide efficient transfer even if the sum of D and J is zero, i.e., if the coupling vanishes in the weak coupling limit. PMID- 11945037 TI - Multidimensional imaging using combined stray field and pulsed gradients. AB - The paper describes an advance in stray field imaging (STRAFI) whereby images of planar samples can be obtained in the stray field of a superconducting magnet without the need for sample rotation. This is achieved by using the static stray magnetic field gradient in combination with pulsed orthogonal gradients. Results of both two- and three-dimensional implementations of the experiment are presented and discussed. An extension to diffusion-weighted imaging is introduced. The technique is expected to prove particularly useful in experiments where high resolution is required in only one direction while lower resolution is acceptable in the orthogonal directions, such as in studies of the drying and curing of paints and varnishes. Arising from the work, a new method for accurately calibrating the radiofrequency pulse width in stray field is found. PMID- 11945038 TI - Determination of hyperfine tensor components from nuclear frequencies at canonical orientations of the g-tensor. AB - The analytical procedure for the determination of all components of the symmetric hyperfine tensor of the I=1/2 nucleus in the g-tensor coordinate system is described, assuming that nuclear frequencies corresponding to the principal directions of the g-tensor and exact values of the external magnetic field (or nuclear Zeeman frequencies) are experimentally available. PMID- 11945039 TI - Complete relaxation and conformational exchange matrix (CORCEMA) analysis of intermolecular saturation transfer effects in reversibly forming ligand-receptor complexes. AB - A couple of recent applications of intermolecular NOE (INOE) experiments as applied to biomolecular systems involve the (i) saturation transfer difference NMR (STD-NMR) method and (ii) the intermolecular cross-saturation NMR (ICS-NMR) experiment. STD-NMR is a promising tool for rapid screening of a large library of compounds to identify bioactive ligands binding to a target protein. Additionally, it is also useful in mapping the binding epitopes presented by a bioactive ligand to its target protein. In this latter application, the STD-NMR technique is essentially similar to the ICS-NMR experiment, which is used to map protein-protein or protein-nucleic acid contact surfaces in complexes. In this work, we present a complete relaxation and conformational exchange matrix (CORCEMA) theory (H. N. B. Moseley et al., J. Magn. Reson. B 108, 243-261 (1995)) applicable for these two closely related experiments. As in our previous work, we show that when exchange is fast on the relaxation rate scale, a simplified CORCEMA theory can be formulated using a generalized average relaxation rate matrix. Its range of validity is established by comparing its predictions with those of the exact CORCEMA theory which is valid for all exchange rates. Using some ideal model systems we have analyzed the factors that influence the ligand proton intensity changes when the resonances from some protons on the receptor protein are saturated. The results show that the intensity changes in the ligand signals in an intermolecular NOE experiment are very much dependent upon: (1) the saturation time, (2) the location of the saturated receptor protons with respect to the ligand protons, (3) the conformation of the ligand-receptor interface, (4) the rotational correlation times for the molecular species, (5) the kinetics of the reversibly forming complex, and (6) the ligand/receptor ratio. As an example of a typical application of the STD-NMR experiment we have also simulated the STD effects for a hypothetical trisaccharide bound to a protein. The CORCEMA theory for INOE and the associated algorithm are useful in a quantitative interpretation of the intensity changes in the ligand in both the STD-NMR and ICS-NMR, provided the identity of the receptor protons experiencing direct RF saturation is known. The formalism presented here is likely to be useful in the design of bioactive ligands to a specific target protein and in the quantitative mapping of binding epitopes and interfaces between molecules in complexes. PMID- 11945040 TI - New line-narrowing effect in triple-quantum resonance in a two-level NMR system. AB - We report a new line-narrowing effect associated with triple-quantum resonance in a two-level NMR system. The experiment was carried out in the rotating frame on (19)F nuclei in Teflon; namely, the magnetization is spin-locked along the RF field and the triple-quantum resonance is induced by the oscillating field perpendicular to the RF field. We observed that the decay time of the triple quantum nutation becomes extraordinarily long at a particular intensity and frequency of the oscillating field. The decay time is about seven times as long as that of the single-quantum nutation and also much longer than that of the magic angle nutation. The mechanism is not interpreted by straightforward analogy to the theory of the current magic angle narrowing. PMID- 11945041 TI - Evaluation and optimization of coherence transfer in high molecular weight systems. AB - For very large proteins in the highest magnetic fields, the large chemical shift anisotropy (CSA) of carbonyl carbon deteriorates coherence transfer efficiency in experiments designed for unambiguous sequential backbone assignment. In this communication, coherence throughput of several TROSY experiments is evaluated. Two new experiments, MP-HNCA and HN(CO)CANH, are also introduced as attractive alternatives for sequential assignment purposes of large proteins with correlation time over 50 ns. Their theoretical coherence transfer efficiencies for the interresidual (13)C(alpha) correlations are significantly better than in recently introduced MP-CT-HNCA and sequential HNCA experiments. The improvement with the new experiments is observed already on 60.8 kDa homodimer of protein Cel6A at 800 (1)H MHz. PMID- 11945042 TI - Adjustable, broadband, selective excitation with uniform phase. AB - An advance in the problem of achieving broadband, selective, and uniform-phase excitation in NMR spectroscopy of liquids is outlined. Broadband means that, neglecting relaxation, any frequency bandwidth may be excited even when the available radiofrequency (RF) field strength is strictly limited. Selective means that sharp transition edges can be created between pure-phase excitation and no excitation at all. Uniform phase means that, neglecting spin-spin coupling, all resonance lines have nearly the same phase. Conventional uniform-phase excitation pulses (e.g., E-BURP), mostly based on amplitude modulation of the RF field, are not broadband: they have an achievable bandwidth that is strictly limited by the peak power available. Other compensated pulses based on adiabatic half-passage, like BIR-4, are not selective. By contrast, inversion pulses based on adiabatic fast passage can be broadband (and selective) in the sense above. The advance outlined is a way to reformulate these frequency modulated (FM) pulses for excitation, rather than just inversion. PMID- 11945043 TI - 1.1-GHz continuous-wave EPR spectroscopy with a frequency modulation method. AB - Continuous-wave EPR spectroscopy using a frequency modulation (FM) scheme was developed. An electronically tunable resonator and an automatic tuning control (ATC) system were used. Using the FM scheme instead of magnetic field modulation, we detected EPR absorption at the first derivative mode. We used a microwave frequency of 1.1 GHz in the present experiment. Similar signal-to-noise ratios were obtained with conventional field modulation and the FM method, and a low quality factor EPR resonator was not necessary to suppress the significant microwave reflection from the resonator. The FM method with a tunable resonator may be an alternative solution to achieving phase-sensitive detection, when the side-effects of magnetic field modulation, such as microphonic noise and mechanical vibration, are detrimental for EPR detection. PMID- 11945044 TI - A rapid and precise probe for measurement of liquid xenon polarization. AB - The relaxation time of liquid (129)Xe is very long (>15 min) and the signal at thermal equilibrium is weak. Therefore, determination of the absolute polarization enhancement of hyperpolarized (129)Xe by direct measurement is tedious. We demonstrate a fast and precise alternative, based on the dipolar field created by liquid hyperpolarized (129)Xe contained in a cylindrical sample tube. The dipolar field is homogeneous in the bulk of the tube and adds to the external field, causing a shift in the Larmor frequencies of all nuclear spins. We show that the frequency shift of the proton in CHCl(3) (chloroform), which dissolves homogeneously in xenon over a fairly broad temperature range, is an excellent probe for (129)Xe polarization. Frequency measurements are precise and the experiment is much faster than by direct measurement. Furthermore the (129)Xe polarization is minimally disturbed since no rf pulses are applied directly to (129)Xe and since chloroform is a fairly weak source of (129)Xe relaxation. The experiments are reproducible and require only standard NMR instrumentation. PMID- 11945045 TI - Signal enhancement in the triple-quantum magic-angle spinning NMR of spins-3/2 in solids: the FAM-RIACT-FAM sequence. AB - We achieve a significant signal enhancement for the triple-quantum magic-angle spinning NMR of a spin-3/2 system, by using an amplitude-modulated radiofrequency field, followed by a selective 90 degrees pulse and a phase-shifted strong rf field, for the triple-quantum excitation, and an amplitude-modulated radiofrequency field for the conversion of triple-quantum coherence to observable single-quantum coherence. The experiment is demonstrated on the (87)Rb NMR of polycrystalline rubidium nitrate. PMID- 11945046 TI - Normal birth. PMID- 11945047 TI - Knowledge, attitudes and practices of trained traditional birth attendants in the Gambia in the prevention, recognition and management of postpartum haemorrhage. AB - OBJECTIVES: to assess the knowledge, attitudes, practices and the potential role of trained Gambian traditional birth attendants (TBAs) in the prevention, recognition and management of postpartum haemorrhage (PPH). DESIGN: a qualitative, reflective approach using semi-structured interviews followed by group discussions. SETTING: poorly-resourced rural villages in The Gambia, West Africa. PARTICIPANTS: 22 trained TBAs and their supervisors from 12 villages. FINDINGS: the TBAs recognised complications such as retained placenta and excessive blood loss and were well aware of the need to refer these women to a health facility quickly. Delay in referral was often due to late call-out of the TBA or lack of transport. Although the TBAs did not know the causes of excessive blood loss, they knew that anaemia was a risk factor for dying from PPH. The TBAs were keen to improve their knowledge and to participate in further training. KEY CONCLUSIONS: although all the TBAs were illiterate, information from training programmes had usually been incorporated into their knowledge and practice. While the local infrastructure remains poor, home deliveries and delayed referrals will continue and interventions for PPH need to be effective at the site of delivery i.e. in the woman's home. These Gambian TBAs have the potential to contribute to the management of PPH in these situations. IMPLICATIONS FOR PRACTICE: these Gambian TBAs could be trained to implement other practices relevant to prevention of PPH in the primary care setting. Linking together and maximising the skills of all health workers is important to reduce PPH mortality in home births in this setting. PMID- 11945048 TI - Developing a theoretical framework on postpartum care from Tanzanian midwives' views on their role. AB - OBJECTIVES: to describe a theoretical framework developed from the views of midwives in relation to provision of systematic postpartum care. DESIGN: qualitative focus group study using grounded theory approach. SETTING: Dar es Salaam, Tanzania. PARTICIPANTS: 49 nurse-midwives in five focus group discussions each having 9-11 participants. FINDINGS: the components of the Basic Social Process of 'Becoming a good resource and support person for the postpartum woman' consisted of 'reflection' as an entry point into the process. Integration, networking, balancing, and dealing with reality, emerged as categories related to process activities. The category of 'defining abilities' required that midwives become aware of their competency and their limitations in reflection and all process activities, so that improvement can be part of 'getting ready', a category that describes what needs to be done at individual and health system level to prepare for systematic postpartum care programmes. The 'caring' category was linked to an outcome of the process 'doing things in the right way', which means providing quality postpartum care. The conditional matrix shows the midwife as an individual affected by several micro and macro conditions. CONCLUSIONS: the proposed theoretical framework can be used in understanding the dynamics of work situations and in assisting midwives to achieve the goal of being good resource and support persons for postpartum women. Interventions for midwives should focus on the major components of the framework but also on the concepts that relate the proposed framework to other central concepts in midwifery and nursing, issues in the theory-practice gap, empowerment, political awareness, involvement in policy making, decision making and dealing with job stress. PMID- 11945049 TI - Postnatal vaginal bleeding problems and General Practice. AB - OBJECTIVES: to investigate women's experiences of problems with vaginal loss from 28 days to three months postnatally and to describe the treatment and referral patterns for women who consult their GP about such problems during the first three months postnatally. DESIGN: a longitudinal questionnaire study of consecutively delivered postnatal women and a report-card survey of GP consultations by women with problems with postnatal vaginal loss. SETTING: two health districts in the south of England. PARTICIPANTS: women delivering in the two health districts during specified recruitment periods in 1995 and 1996. For the GP study, with her consent, the GP returned an anonymous registration card for each woman presenting. FINDINGS: in the survey of women, 20% (64/325) reported problems with postnatal loss occurring between 28 days and three months after the birth. Around a half of these consulted a GP. The GP study was disappointing in that only 26% (30/115) of practices agreed to take part and 16% (18/115) returned notification cards. Forty-eight women were included from 18 practices. The most common presenting symptoms were excessive bleeding (29/48; 60%) and prolonged bleeding (26/48; 54%). The commonest form of treatment was antibiotics alone (15/48; 31%) but 12 women (25%) were neither treated nor referred. Referral (n=19) was for hospital admission, out-patient appointment or direct referral for an ultrasound scan. KEY CONCLUSIONS AND IMPLICATIONS FOR PRACTICE: morbidity related to abnormalities of postnatal vaginal fluid loss (lochia) has been shown to be significant, yet nothing was known about the outcome of related GP consultations during the first three months postnatally. A variety of treatment and referral patterns were revealed, highlighting the need for a systematic review of the literature on the management of secondary postpartum haemorrhage. Health care workers need to be aware of the significant morbidity experienced by postnatal women in relation to their lochial loss. PMID- 11945050 TI - What women from an Islamic background in Australia say about care in pregnancy and prenatal testing. AB - OBJECTIVE: to examine satisfaction with care and services in relation to antenatal care and prenatal testing and to present what women say about what can be done better to improve antenatal care for women from an Islamic background. DESIGN: in-depth interviews of women's perceptions and experiences of care received relating to prenatal testing and antenatal care. SETTING: Melbourne Metropolitan Area, Victoria, Australia. PARTICIPANTS: 15 women of Islamic background who are now living in Melbourne. FINDINGS: in general, women had positive experiences with care relating to antenatal care and prenatal testing in Australia. This is particularly so when they compared care in Australia with that of their own country. However, women indicated several issues of concern where they were dissatisfied and they believed need to be improved for pregnant women from an Islamic background. Firstly, there was a lack of sufficient communication between health care providers and the women. This was not only due to a language problem, but also a lack of cultural appreciation among health care providers. Secondly, women identified the issue of gender of health care providers as important; women stated clearly their need to have female doctors for their care. CONCLUSIONS: the findings of this study have implications for antenatal care and prenatal testing services in Australia and elsewhere. Women provided several suggestions for the improvement of care including the need for sufficient information of prenatal testing and antenatal care and the need for culturally sensitive services. In providing services for women of an Islamic background, it is imperative that health care providers take into account individual women's preferences and personal circumstances and go beyond an assumption based on women's religion and ethnicity. PMID- 11945051 TI - The significance of time factors in cerebral palsy litigation. AB - OBJECTIVE: to demonstrate the significance of the lengths of time taken to initiate and conclude litigation concerning cerebral palsy. DESIGN: documentary analysis of 142 closed legal files (92 Scottish; 50 English) relating to births from 1980-1996. Claim outcome, and the dates of birth, legal notification and conclusion of the claim, were analysed using Microsoft Excel. SETTING: health service legal offices in Scotland and England. FINDINGS: successful claims were raised more quickly (on average two years compared with 3.6 years for unsuccessful claims). They also took longer to be decided (six years compared with three years). On average the children of successful and unsuccessful litigants were, respectively, 7.8 and 6.6 years old by the time the claim was decided. Over the period studied there was a steady reduction in the length of time taken to initiate litigation, although successful claims were on average raised more quickly. Overall success rate was 27%. CONCLUSIONS/IMPLICATIONS FOR PRACTICE: most cerebral palsy claims do not satisfy the requirements of the law of medical negligence. Lengthy periods of time are involved irrespective of outcome: while successful litigants continue to sue more quickly, their claims take longer to be resolved. The extensive period from birth to closure of claim, and uncertainty over the legal outcome, may cause significant distress for practitioners and parents of children with cerebral palsy. Knowledge of the likely duration of the legal process can help practitioners and the families involved to come to terms with this important feature of litigation. Health service claims managers may also be helped in terms of understanding the more likely duration (and therefore cost implications) of cerebral palsy claims. For successful litigants there are questions about the justice of having to wait, on average, nearly eight years for compensation. PMID- 11945052 TI - A critical ethnographic approach to facilitating cultural shift in midwifery. AB - OBJECTIVE: to improve understanding of local midwifery morale, inform development and reorganisation of a maternity unit, and enhance midwifery involvement in strategic planning. PARTICIPANTS: a randomised stratified sample of 20 midwives working in a UK National Health Service (NHS) hospital and its surrounding community area. METHOD: within a critical ethnographic framework, focus groups were tape-recorded and transcribed, and analysed using a thematic content analysis approach. FINDINGS: key areas affecting midwifery morale were identified, in particular staffing levels, working relationships and organisational issues. One year later, despite many changes having taken place, midwifery morale was still low but participants were more politically analytical of, and actively involved in changing their situation. The findings of the study indicate that there are complex and long-standing cultural inhibitions to the effective development of midwifery care but, if these are made explicit through a planned collaborative process, such as in this study, a process of cultural shift can be seen to begin. IMPLICATIONS FOR PRACTICE: focus groups can be a useful tool in moving midwifery culture forward within a local context. PMID- 11945053 TI - Parents' experience of early discharge from hospital after birth in Sweden. AB - OBJECTIVE: to investigate the factors that influence the experience of mothers and fathers when they have chosen to return home, earlier than is the normal routine, following the birth of their baby. DESIGN: a qualitative study, using open interviews, was undertaken. The text of the transcripts was coded and categorised according to the grounded theory method using constant comparative analysis. SETTING: interviews were carried out with 12 parents, six mothers and six fathers, individually in their own homes. They had all left a maternity/family ward at the Helsingborg Hospital in southern Sweden within 26 hours of birth whereas the normal discharge time is 72 hours. MEASUREMENTS AND FINDINGS: 'a sense of security' was the core category. Achieving a sense of security linked to informed choice for early discharge appeared to be dependent on the following categories: (l) the midwives' empowering behaviour; (2) affinity within the family; (3) the parents' right to autonomy/control; (4) physical well being. There appears to be an inner connection between each of these categories. KEY CONCLUSIONS: the midwife's empowering behaviour supports the parents' sense of security and encourages their informed choice of earlier discharge after birth. When the mothers' and babies return home it strengthens the affinity within the family and the father's sense of participation. PMID- 11945054 TI - Finnish women's experiences of antenatal care. AB - OBJECTIVE: to explore and describe women's experiences of antenatal care. DESIGN: semi-structured interviews, dialogical interviews and non-participant observation in two phases both during and after pregnancy were analysed according to Colaizzi's phenomenological method. SETTING: four maternity clinics in one rural and one urban primary health care centre and one outpatient maternity clinic at a central hospital in Finland. PARTICIPANTS: in the first phase a purposive sample of nine women were interviewed at 36 weeks gestation, three weeks, three months and two and a half years after birth. In the second phase, data were collected for further breadth and depth and 31 women, who were going through different stages of pregnancy, delivery and the puerperium participated. Data were collected by means of interviews supplemented by non-participatory observation. In the total sample of 40 Finnish women, one half was primigravidae and the other half multigravidae. FINDINGS: a main thread running through the women's experiences of antenatal care were their needs and wishes that concerned the health of the unborn baby, but also the health and dignity of themselves and their family. The pregnant women wanted to share their pregnancy and childbirth story in a confident relationship based on humaneness and interest in a peaceful atmosphere. They needed protection safeguarded through scientifically based and humane surveillance, and professional competence was expected. They wanted to continuously participate in the new situations through knowledge in dialogues and they wanted to involve their family in the care on their own terms. The childbirth preparation groups had an important multidimensional role in the care. IMPLICATIONS FOR PRACTICE: the findings challenge the antenatal care that so far has been focused on the physical health, the needs of the primigravida and a blurred family perspective. The family perspective implies knowing both the pregnant woman and her partner in terms of the pregnancy, the birth and a new parenthood. Humane, scientifically based perinatal care can be developed by innovations from these findings, especially considering the multidimensional role of the parent groups. PMID- 11945055 TI - A review of the literature on debriefing or non-directive counselling to prevent postpartum emotional distress. AB - BACKGROUND: childbirth generates powerful emotions and may lead to the development of symptoms of depression, anxiety, and trauma in some women. Debriefing and non-directive counselling have been used as early interventions to reduce the prevalence of depression and post-traumatic stress. METHODS: a review of the literature was conducted to describe the current state of knowledge on the effectiveness of a single debriefing session or non-directive counselling session to reduce depression and trauma symptoms in women following birth. FINDINGS: a total of three studies reported in four papers examined the use of debriefing or non-directive counselling to prevent or reduce psychological morbidity following birth. The two largest RCTs indicate that a single debriefing session with the woman whilst in the postnatal ward is of no statistically significant value in reducing psychological morbidity and may even be harmful. In contrast, women reported that an opportunity to talk with someone about the birth was helpful in facilitating recovery. CONCLUSION: there is insufficient evidence to draw conclusions about the effectiveness of debriefing following childbirth, primarily because it is unclear if a standardised debriefing intervention was used. Future research should clearly describe the intervention and test alternative interventions; measure a broader range of outcomes including trauma symptoms; use inclusion criteria that acknowledge the complex contributing factors to depression and trauma; and examine the value of including the woman's partner (or significant other) in the debriefing or counselling session(s). Future studies should investigate the timing or place of the intervention, the provision of more than one opportunity to discuss the birth, and target the intervention to women who are more likely to develop trauma symptoms or post-traumatic stress disorder. PMID- 11945057 TI - Censorship of scientific publications: a bad idea. PMID- 11945058 TI - Can "negative" be positive? PMID- 11945059 TI - Evolution of adenoviruses as gene therapy vectors. PMID- 11945060 TI - Continuous intravascular secretion of endostatin in mice from transduced hematopoietic stem cells. AB - Endostatin, a 20-kDa carboxy-terminal fragment of collagen XVIII, is the leading member of a class of physiologic inhibitors of angiogenesis with potent antitumor activity. Repeated subcutaneous administration of recombinant endostatin in mice led to permanent regression of established tumors to a microscopic dormant state and prompted the initiation of human clinical trials. However, a discrepancy remained unresolved: sustained tumor regression has only been observed with a non soluble, precipitated form of recombinant endostatin produced in bacteria. To shed light on this question and establish a model of systemic anti-angiogenic gene therapy of cancer that may surmount obstacles in protein production and delivery, we transduced murine hematopoietic stem cells with a retrovirus encoding a secretable form of endostatin. Despite continuous, high-level secretion of endostatin in the vasculature of all transplanted mice, we detected neither inhibition of in vivo neoangiogenesis nor antitumor activity. Resolution of this paradox may come from human trials of endostatin now underway. PMID- 11945061 TI - Unfulfilled promise of endostatin in a gene therapy-xenotransplant model of human acute lymphocytic leukemia. AB - Retroviral transduction of hematopoietic stem cells (HSCs) offers an attractive strategy for treating malignancies that home to the marrow. This approach should therefore be of interest for evaluating the therapeutic activity of anti angiogenic agents on hematopoietic malignancies whose growth has been associated with enhanced angiogenesis. A variety of studies have indicated endostatin to be a potent anti-angiogenic agent both in vitro and in vivo, and a human malignancy that might be sensitive to endostatin is human B-lineage acute lymphoblastic leukemia (B-ALL). The demonstrated ability of human B-ALL cells to engraft the marrow of immunodeficient mice suggested the potential of this system for testing an endostatin delivery strategy using co-transplanted non-obese diabetic scid/scid (NOD/SCID) HSCs engineered to express endostatin. Here we show that, in spite of their mutant scid gene, NOD/SCID HSCs can be transduced with an endostatin-encoding retrovirus at efficiencies that result in a several-fold increase in endostatin serum levels in transplanted recipients. However, this did not alter the regrowth of co-transplanted human B-ALL blasts. These findings validate this gene transfer approach for investigating effects of novel therapeutics on primary human malignant cells that engraft NOD/SCID mice and question the utility of native endostatin for controlling human B-ALL in vivo. PMID- 11945062 TI - Correct integration mediated by integrase-LexA fusion proteins incorporated into HIV-1. AB - Fusion of wild-type or truncated integrase to a sequence-specific DNA-binding protein, such as the Escherichia coli LexA repressor, results in an integration bias toward the recognition site of the DNA-binding protein in vitro. Integrase defective HIV-1 could become integration-competent by supplying the fusion protein in trans. To understand the mechanism of complementation, the virus-host DNA junctions of cells infected with the integrase-LexA containing virus were sequenced. The characteristic hallmarks of wild-type integration were present, a 5'-TG/CA-3' at the ends of the viral sequence and a 5-bp direct repeat in the immediately flanking cellular DNA. Experiments were also carried out to determine the mechanism by which the amino- or carboxy-terminal truncated integrase fused to LexA restored integration to the integrase-mutant viral clone. Complementation experiments using purified fusion proteins in vitro, or viruses encoding a C terminal truncated integrase and containing various fusion proteins in trans, indicated that the truncated integrase-LexA proteins are inactive per se and they restore integration by forming mixed multimers with the virally encoded mutant integrase. Correct integration of retroviral DNA by the in trans method illustrates the feasibility of introducing integrase fusion proteins into retroviral vectors to achieve site-directed integration without interfering with the attributes of the integration reaction. PMID- 11945063 TI - Recombinant adeno-associated virus serotypes 2- and 5-mediated gene transfer in the mammalian brain: quantitative analysis of heparin co-infusion. AB - Recombinant adeno-associated viruses (rAAVs) are among the most promising vectors for gene delivery into the CNS. However, a major hurdle for gene transfer to the mammalian brain is to achieve high transduction levels in target cells beyond the immediate injection site. Therefore, building upon the optimization of injection parameters on which we have recently reported, it is important to define additional methods to increase the volume of distribution. Here, we establish an optimal heparin concentration, and show that co-injection of heparin together with rAAV2 leads to a significantly higher and more homogeneous distribution of transduced cells. In contrast, the diffusion pattern of rAAV serotype 5 differs from that of rAAV2, in that its distribution is less homogeneous, more variable, and patchy. Furthermore, this study illustrates the influence of receptor binding on the expression pattern following injection of rAAV in the CNS. In addition to improvements in expression cassettes and viral titers and the use of very slow infusion rates, gene transfer studies in the CNS where the goal is to obtain widespread transduction should consider co-injecting the viral vector rAAV2 with heparin to maximize transduction efficiency and viral spread. PMID- 11945064 TI - Lentiviral transduction of P140K MGMT into human CD34(+) hematopoietic progenitors at low multiplicity of infection confers significant resistance to BG/BCNU and allows selection in vitro. AB - Lentiviral vectors may improve hematopoietic stem cell (HSC) gene transfer because of their enhanced ability to transduce nondividing cells. However, many studies report efficient transduction only at high multiplicities of infection (MOI). This study reports efficient transduction of human CD34(+) cells with a drug resistance gene allowing post-transduction selection using lentivirus under low-MOI conditions that did not require cytokine stimulation or viral concentration. We used the P140K methylguanine-DNA-methyltransferase mutant (P140K MGMT) as the gene insert into a second-generation lentiviral backbone and triple-plasmid transfection to generate vesicular stomatitis virus (VSV)-G protein-pseudotyped virus. The P140K MGMT gene product, O(6)-alkylguanine-DNA alkyltransferase (AGT), provides protection from the therapeutic drug combination of 1,3-bis(2-chloroethyl)-1-nitrosourea (BCNU) and the wild-type AGT inhibitor O(6)-benzylguanine (BG). Low-speed spinoculation enhanced transduction more than addition of Polybrene or multiple virus exposures. Addition of cytokines was not required. Low-MOI transduction (< or =1) of human CD34(+) and CD34(+) lin(-) cells with P140K MGMT lentivirus resulted in an average 41% and 89% gene transfer rate as assessed by PCR, respectively, and concordant AGT expression that conferred substantial clonogenic survival advantage after BG/BCNU treatment. During in vitro drug selection, 87% of surviving CD34(+) cell-derived colony forming units (CFU) were transduced. This work shows the potential utility of lentiviral vectors for drug resistance gene transfer to HSCs for the purpose of in vivo selection and marrow protection. Because drug selection will enrich for transduced progenitors, high MOI can be avoided, improving the safety profile of lentiviral gene transfer. PMID- 11945066 TI - In vivo gene delivery to synovium by lentiviral vectors. AB - The delivery of anti-arthritic genes to the synovial lining of joints is being explored as a strategy for the treatment of rheumatoid arthritis. In this study, we have investigated the use of VSV-G pseudotyped, HIV-1-based lentiviral vectors for gene delivery to articular tissues. Recombinant lentivirus containing a beta galactosidase/neomycin resistance fusion gene under control of the elongation factor (EF) 1alpha promoter efficiently transduced human and rat synoviocytes and chondrocytes in cell culture. When directly injected into the knees of rats, this vector transduced synovial lining cells, but not other articular tissues such as cartilage. We also constructed a lentiviral vector containing the human interleukin-1 receptor antagonist (IL1RA) cDNA and examined transgene expression in vitro and in vivo following injection into the knee joints of rats. In immunocompetent animals, intra-articular IL1RA expression was high and persisted, at a sharply declining rate, for approximately 20 days. In immunocompromised rats, however, lentivirus-mediated intra-articular expression of human IL1RA was found to persist for at least 6 weeks. Extra-articular expression of the transgene was minimal. These results indicate that lentiviral vectors are capable of efficient in vivo gene transfer to synovium and merit further investigation as a means of providing long-term expression for gene-based treatments of arthritis. PMID- 11945065 TI - Cytotoxic immune response blunts long-term transgene expression after efficient retroviral-mediated hepatic gene transfer in rat. AB - Vectors derived from oncoretroviruses can transduce a small proportion of hepatocytes when injected in the regenerating liver. Transgene expression may be sustained for months without immune response. In striking contrast, we observed a rapid extinction when the intravenous injection of a high input of nuclear beta galactosidase (beta-gal) expression vector, one day after partial hepatectomy, led to a significant proportion of transduced cells in the liver. Extinction was associated with liver inflammation on tissue sections and appearance of antibodies against the transgene product, while vector genomes became undetectable in liver tissue by PCR. These observations suggested the elimination of transduced cells by an immune response. Transgenic rats tolerant for cytoplasmic beta-gal, or normal rats depleted in CD8 T lymphocytes, steadily expressed the beta-gal vector. In the spleen of normal rats, we detected cytotoxic cells directed against cells expressing beta-gal after the injection of the beta-gal vector. In jaundiced Gunn rats deficient in bilirubin glucuronosyl transferase (BGT1) and treated with a human BGT1 cDNA expression vector, we observed the same kinetics of extinction as well as the appearance of anti-BGT1 antibodies. This study demonstrates that retrovirus-mediated gene transfer may induce cytotoxic T lymphocytes specifically directed against transgene-expressing cells. PMID- 11945067 TI - Improved treatment of pancreatic cancer by IL-12 and B7.1 costimulation: antitumor efficacy and immunoregulation in a nonimmunogenic tumor model. AB - Ductal pancreatic adenocarcinoma is one of the commonest and most lethal cancers in the Western world. Unfortunately, recent advances in diagnostics, staging, and therapy in pancreatic carcinoma have not resulted in significant improvements in long-term survival. We have previously shown that adenovirus (Ad)-mediated coexpression of interleukin-12 (IL-12) and the costimulatory molecule B7.1 is extremely efficient in inducing regression of highly immunogenic transplanted and nontransplanted tumors. Here, we examined the antitumor efficacy of IL-12- and B7.1-based immunotherapy against a nonimmunogenic murine model of ductal pancreatic cancer. Compared with AdIL-12 treatment alone, single intratumoral injection of AdIL-12/B7.1 led to a prolonged immune response and mediated complete regression in 80% of treated animals. After rechallenge with parental tumor cells, 70% of cured mice remained tumor-free, suggesting that protective immunity had been induced. The antitumoral response was associated with upregulation of H-2K(b) and Abcb2 expression, whereas other components of the proteasome (Abcb3, Psmb9, and Psmb8) were not affected. These data indicate that upregulation of the antigen presentation machinery by AdIL-12/B7.1 may be a therapeutic rationale for nonimmunogenic, therapy-resistant pancreatic cancer. PMID- 11945068 TI - Functional evidence of CFTR gene transfer in nasal epithelium of cystic fibrosis mice in vivo following luminal application of DNA complexes targeted to the serpin-enzyme complex receptor. AB - Molecular conjugates that target the serpin-enzyme complex receptor transfer the cDNA encoding human cystic fibrosis transmembrane conductance regulator (CFTR) to the nasal epithelium of cystic fibrosis mutant mice. These complexes effect partial correction of the chloride transport defect as assessed by in vivo nasal potential difference measurements, produce immunohistochemical staining for CFTR, and restore expression of nitric oxide synthase-2 (NOS-2), which is downregulated in the epithelium of mice and humans with cystic fibrosis. Complexes that lack the receptor ligands were ineffective, so receptor access was essential. Mice treated with receptor-targeted lacZ showed beta-galactosidase expression in epithelial cells and submucosal glands, but no electrophysiologic correction or NOS-2 expression, so simply accessing the serpin-enzyme complex receptor was not sufficient to produce the observed electrophysiologic or immunohistochemical changes. Correction of the cAMP-stimulated chloride transport was dose related at days 7 and 12 after complex administration, but, for most animals, nasal potential difference had returned to baseline by day 18. Molecular conjugates targeting the serpin-enzyme complex receptor, used to compact plasmid DNA, hold promise for gene therapy of cystic fibrosis. PMID- 11945069 TI - Dimerizer-induced proliferation of genetically modified hepatocytes. AB - Stable gene transfer to the liver by viral vectors is inefficient. In an attempt to stimulate expansion of retrovirally transduced hepatocytes, we employed a synthetic drug (AP20187) that can reversibly dimerize and activate fusion proteins that contain a growth factor receptor signaling domain. Signaling domains derived from receptors for interleukin-6 (gp130), hepatocyte growth factor (c-met), epithelial growth factor (EGF-R), and thrombopoietin (mpl) triggered monkey hepatocytes to enter the cell cycle. However, mitosis occurred only upon activation of the gp130 and c-met signaling domains. Primary mouse hepatocytes expressing the gp130 fusion proliferated transiently in response to AP20187. AP20187-triggered activation of gp130 also stimulated the selective (>2 fold) expansion of retrovirally transduced hepatocytes in vivo, as shown by immunohistochemical staining and quantitative proviral DNA analysis. Drug inducible in vivo expansion of genetically modified hepatocytes may have potential applications in hepatic gene transfer or in liver repopulation by transplanted hepatocytes or their progenitors. (c)2002 Elsevier Science (USA). PMID- 11945070 TI - Infectious delivery of 120-kilobase genomic DNA by an epstein-barr virus amplicon vector. AB - It has been shown in a wide variety of contexts that persistent gene expression can best be obtained by using the genomic locus of a transgene. However, the size of most genomic loci precludes their use in current viral gene therapy vectors. Large transgene capacity and extrachromosomal persistence make Epstein-Barr virus (EBV) a promising vector to deliver genomic transgenes for gene therapy. We constructed an EBV amplicon vector that contains the EBV lytic origin of replication, the terminal repeats for viral packaging, and the EBV latent origin of replication for episomal persistence. This vector was able to deliver inserts of 60-123 kb to B-cell lines in culture in three steps. First, clonal packaging cells lines were generated that produce infectious amplicons at a titer of approximately 3-4x10(6) transducing units/ml after concentration. Second, we show infectious vector delivery to the Loukes B-cell line and three different EBV immortalized lymphoblastoid cell lines. This infectious delivery system was 2000 times more efficient than transfection in B cells. Third, clonal cell lines from infection of Loukes contained persistent episomes of recircularized infectious vector. This first demonstration of infectious delivery of 120 kb of genomic DNA shows the potential of this high-capacity vector system. PMID- 11945071 TI - Efficacy of gene therapy for a prototypical lysosomal storage disease (GSD-II) is critically dependent on vector dose, transgene promoter, and the tissues targeted for vector transduction. AB - Lysosomal storage diseases are an intriguing target for gene therapy approaches, as transduction of a "depot" organ with a transgene encoding a lysosomal enzyme can be followed by secretion, systemic distribution, downstream uptake, and lysosomal targeting of the enzyme into non-transduced tissues. These benefits are of utmost importance when considering gene therapy approaches for glycogen storage disease type-II (GSD-II). GSD-II is a prototypical lysosomal storage disorder caused by lack of intralysosomal acid alpha-glucosidase (GAA) activity. Lack of GAA can result in a proximal limb myopathy and respiratory and cardiac failure, each due to abnormal glycogen accumulation in the skeletal muscles or cardiac tissues, respectively. After converting the liver into a "depot" organ, we found that intravenous injection of the [E1-,polymerase-]AdGAA vector allowed for hepatic secretion of GAA over an at least 20-fold dosage range. We noted that very low plasma GAA levels (derived from hepatic secretion of GAA) can allow for GAA uptake by muscle tissues (skeletal or cardiac), but significantly higher plasma GAA levels are required before glycogen "cross-correction" can occur in these same tissues. We also demonstrated that liver-specific enhancer/promoters prolonged GAA transgene expression from persistent [E1-,polymerase-] adenovirus based vector genomes for at least 180 days, and significantly diminished the amounts of neutralizing anti-GAA antibodies elicited in this animal model. Finally, we demonstrated that skeletal muscles can also serve as a "depot" organ for GAA secretion, allowing for secretion of GAA and its uptake by noninfected distal tissues, although glycogen reductions in non-injected muscles were not achieved by the latter approach. PMID- 11945072 TI - Secretin-mediated gene delivery, a specific targeting mechanism with potential for treatment of biliary and pancreatic disease in cystic fibrosis. AB - Gene therapy directed to the gastroenterological manifestations of cystic fibrosis (CF) would ideally be administered systemically. Such delivery would require efficient targeting at the cellular level to achieve a safe and effective therapy. Here we describe gene delivery using the secretin receptor (SR) as a basolateral target specific to the biliary and pancreatic epithelia affected in CF patients. We describe here targeting of a polycation-based nonviral gene delivery vector and retargeting of an adenoviral vector to cells expressing the SR in vitro. We were able to transfect cells expressing the SR up to 10-fold more efficiently than those not expressing the SR with a targeted polycation, SecGGC lPEI. This targeting effect was secretin-specific and substantially reduced by competing secretin. SR-retargeted adenovirus transduced SR-expressing cells at more than sixfold higher levels than adenovirus alone. The SR may be an effective target for targeting systemically applied viral and nonviral gene delivery constructs to disease-affected tissues in CF patients. PMID- 11945073 TI - Enhancement of transgene expression by combining glucocorticoids and anti-mitotic agents during transient transfection using DNA-cationic liposomes. AB - The anti-mitotic drugs colchicine and paclitaxel increase transfection efficiency of cationic liposomes. Using combined lipid-mediated transfection with anti mitotic agents for gene therapy of cancer has been limited due to the likely development of multi-drug resistance (MDR). We treated human cancer cell lines and normal liver cells with glucocorticoids in combination with the antimitotics paclitaxel or colchicine before transient, cationic lipid-mediated transfection. Colchicine and paclitaxel each enhanced transgene expression in several cell lines. Moreover, glucocorticoid, combined with paclitaxel or colchicine, significantly increased reporter gene expression above that seen in cells treated with each drug alone. P-glycoprotein (PGP), a drug exporter encoded by ABCB1, exports both paclitaxel and colchicine. To determine the influence of PGP in colchicine- or paclitaxel-mediated enhancement of transgene expression, cells were treated with a histone deacetylase inhibitor, trichostatin A (TSA), known to induce ABCB1 expression, before treatment with colchicine or paclitaxel. TSA significantly reduced colchicine-mediated increases in reporter gene expression. Addition of glucocorticoid to colchicine pretreatment significantly attenuated TSA-mediated inhibition of colchicine-induced increases in transgene expression. TSA accelerated and glucocorticoid blocked export of rhodamine 123, a molecule known to be exported by PGP. The glucocorticoid/paclitaxel combination also increased reporter gene expression in BE(2)C cells, which constitutively express high levels of PGP. Thus, the degree of enhancement of transgene expression mediated by these anti-mitotics seems to be dependent on PGP activity. Glucocorticoids augment colchicine- or paclitaxel-mediated enhancement of transgene expression most likely by reducing drug egress through PGP. PMID- 11945074 TI - Importance of lateral and steric stabilization of polyelectrolyte gene delivery vectors for extended systemic circulation. AB - Gene therapy for systemic diseases requires intravenous administration, but existing vectors are not suitable for systemic delivery, often showing rapid elimination from the bloodstream that restricts potential transfection sites to "first-pass" organs. To develop long-circulating vectors, here we have compared polyplexes containing DNA and poly-L-lysine (PLL) or polyethylenimine (PEI), surface-modified with either monovalent polyethylene glycol (PEG) or multivalent copolymers of N-(2-hydroxypropyl)methacrylamide (PHPMA), correlating their biophysical properties with their distribution following intravenous injection. A key difference between the two types of coating is the introduction of lateral stabilization by surface attachment of multivalent PHPMA, in addition to the steric stabilization provided by both types of polymers. The alpha-half-life for bloodstream clearance of polycation/DNA polyplexes (typically <5 minutes in mice) could be extended using multivalent PHPMA coating to >90 minutes. We found that the dose administered, as well as the amount and molecular weight of the coating PHPMA, had important effects on circulation properties. Multivalent PHPMA coating allows, for the first time, considerably extended circulation time using polyplex systems-a prerequisite for systemic gene delivery. PMID- 11945075 TI - Highly efficient retroviral gene transfer based on centrifugation-mediated vector preloading of tissue culture vessels. AB - Efficient retroviral gene transfer into primary cells is a prerequisite for various gene therapeutic strategies. We have developed a transduction protocol based on the preloading of tissue culture vessels with retroviral particles by low-speed (1000g) centrifugation. We show that vector-preloaded tissue culture vessels allow highly efficient gene transfer into various target cells. We obtained transduction rates of up to 85% for primary T lymphocytes after just a single round of transduction. Under clinically relevant conditions using a vector developed for suicide gene therapy and produced under good manufacturing practice (GMP) conditions, the described method allowed generation of large numbers (>2x10(9)) of gene-modified T cells. The preloading concept ensures transduction of target cells in their optimal growth medium regardless of the medium used for vector production. This facilitated highly efficient gene transfer into quite different target cells such as CD34(+) and AC133(+) bone marrow progenitor as well as mesenchymal stem cells. The presented method combines high gene-transfer rates with a great potential for standardization in accordance with GMP guidelines and is consequently well suited for both research and clinical applications. (c)2002 Elsevier Science (USA). PMID- 11945076 TI - Design of HIV vectors for efficient gene delivery into human hematopoietic cells. AB - Vectors derived from human immunodeficiency virus (HIV) hold promise for efficient gene delivery into human hematopoietic cells. In this study, HIV vectors containing different combinations of cis-acting elements, including the HIV central flap sequence, and the woodchuck posttranscriptional regulatory element (WPRE) in combination with two different promoters, were used to transduce primary human lymphocytes and cord blood CD34+ progenitor cells. The effect of these elements on the transduction efficiency and transgene expression was systematically evaluated. The results demonstrate that with the combination of flap, WPRE sequences, and the promoter derived from spleen focus-forming virus (SFFV), a foreign gene can be efficiently delivered into primary human T lymphocytes and cord blood CD34+ cells. The study establishes the parameters for proper vector design to efficiently deliver foreign genes into human hematopoietic cells. PMID- 11945077 TI - Prospective function of placental leptin at maternal-fetal interface. AB - Leptin is an endocrine and a growth factor which is important for regulation of body fat, feeding, and energy homeostasis. The anti-obesity function of leptin has been recently extended to reproduction, puberty and pregnancy as an endocrine signal to the hypothalamus. Leptin controls the functional integrity of the feto placental unit thereby maintaining pregnancy by virtue of its immunomodulatory property via T lymphocytes or other proto-oncogenes. Dysregulation of autocrine/paracrine function of leptin at feto-placento-maternal interface may be implicated in the pathogenesis of recurrent miscarriage gestational diabetes, pre eclampsia and intra-uterine fetal growth retardation including disturbance of fetal bone turnover. This review will focus on the role of leptin in normal and abnormal pregnancy and fetal growth. PMID- 11945078 TI - Early placental ontogeny in the mouse. AB - Fundamental to placental morphogenesis is union between the allantois and the chorion, two tissues initially separated in the conceptus. Results of previous studies in the mouse have suggested that chorio-allantoic union is driven by the developmental maturity of the allantois and involves molecular interactions between Vascular Cell Adhesion Molecule (VCAM-1) in the allantois and alpha4 integrin in the chorion. Little more is known about the cellular and/or molecular control of this important morphogenetic event in any species.Gross, histological, microsurgical and immunohistochemical analyses in the mouse conceptus revealed that placental ontogeny took place in three major steps. The first, chorio allantoic contact, was not enduring and was mediated by the allantois' mesothelial surface and the mesodermal component of the chorion. Modest amounts of VCAM-1 were found in distal allantoic mesothelium, whilst levels of alpha4 integrin were high throughout chorionic mesoderm. The second step, chorio allantoic fusion, was more enduring. During this time, the distal allantoic region contained maximal levels of VCAM-1, and all allantoises had expanded far enough to reach the posterior chorion from where they spread toward a central chorionic depression. The last step, breakdown of chorio-allantoic fusing surfaces, was dependent upon chorio-allantoic fusion and resulted in the intimate juxtaposition of allantoic endothelium and chorionic ectoderm, possibly as a result of VCAM-1-mediated interactions. The umbilical connection was thereafter fixed at its perimeter to the chorionic surface by large amounts of VCAM-1 in disto-lateral allantoic mesothelium and alpha4-integrin in the remaining peripheral mesodermal component of the chorion.Thus, chorio-allantoic union is highly regulated, taking place in multiple steps. It is dependent upon the developmental maturity of distal allantoic mesothelium and involves the mesodermal component of the chorion. Breakdown of fusing surfaces enables penetration of the allantoic vasculature into the chorion. These findings provide a secure developmental foundation in which to elucidate the genetic control of early placentation. PMID- 11945079 TI - Placental expression of VEGF, PlGF and their receptors in a model of placental insufficiency-intrauterine growth restriction (PI-IUGR). AB - Placental development requires adequate and organized interaction of vascular growth factors and their receptors, including vascular endothelial growth factor (VEGF) and placental growth factor (PlGF). Both VEGF and PlGF, acting through the tyrosine kinase receptors VEGFR-1 and VEGFR-2, have been implicated in playing a role in ovine placental vascular development. The present studies describe the placental expression of components of the VEGF family at two maturational time points (55 and 90 days post coitus, dpc) in a hyperthermic-induced ovine model of placental insufficiency-intrauterine growth restriction (PI-IUGR). Both caruncular and cotyledonary VEGF and PlGF mRNA concentration increased with gestational age (P< 0.05), whereas only cotyledonary VEGF and PlGF protein concentration increased over gestation (P< 0.002). At 55 dpc, VEGF mRNA concentration was elevated in hyperthermic (HT) ewes, compared to control thermoneutral (TN) animals (TN; 0.52+/-0.08 vs HT; 1.27+/-0.17 VEGF/GAPDH, P< 0.001). At 90 dpc, expression of PlGF and VEGF mRNA was not altered by the HT treatment. Both TN cotyledonary VEGFR-1 and VEGFR-2 mRNA expression levels rose significantly over the period studied (P< 0.05 and P< 0.01 respectively). Receptor mRNA concentration in HT cotyledonary tissue was significantly reduced at 90 dpc (VEGFR-1; TN 0.21+/-0.02 vs HT 0.11+/-0.01 VEGFR-1/actin, P< 0.05, VEGFR-2; TN 0.18+/-0.05 vs HT 0.07+/-0.01 VEGFR-2/actin, P< 0.01). Soluble VEGFR 1 (sVEGFR-1) mRNA was not detected in these tissues. These alterations in growth factor and growth factor receptor mRNA expression, as a result of environmental heat stress early in placental development, could impair normal placental vascular development. Furthermore, alterations in VEGF, VEGFR-1 and VEGFR-2 mRNA expression, during the period of maximal placental growth, may contribute to the development of placental insufficiency, and ultimately intrauterine growth restriction. PMID- 11945080 TI - Subcellular localization of PP5/TFPI-2 in human placenta: a possible role of PP5/TFPI-2 as an anti-coagulant on the surface of syncytiotrophoblasts. AB - Placental protein 5 (PP5)/tissue factor pathway inhibitor-2 (TFPI-2), a serine proteinase inhibitor, is homologous to tissue factor pathway inhibitor (TFPI) and commonly found in peripheral blood of pregnant woman. Although TFPI is well known to be synthesized primarily in endothelium and to play an important role in regulation of the extrinsic pathway of blood coagulation, the function of PP5/TFPI-2 remains unclear. Our previous report demonstrated that PP5/TFPI-2 expression is ubiquitous and extremely high in growing placental tissue. Using newly generated polyclonal anti-PP5/TFPI-2 antibody, and by immunohistochemistry and immunoelectromicroscopy, we examined precise localization of PP5/TFPI-2 in placenta especially in syncytiotrophoblasts, which had been shown to produce PP5/TFPI-2 mRNA by our previous study using in situ hybridization. Immunoelectromicroscopy revealed PP5/TFPI-2 localizing on the surface of the microvilli and the membrane of endoplasmic reticulum of syncytiotrophoblasts. To confirm the cell surface association of PP5/TFPI-2, placental villi were incubated with heparin and resultant soluble fraction was analysed by Western blotting. Heparin liberating PP5/TFPI-2 from villi suggested that PP5/TFPI-2 might be retained on the microvilli surface through the binding to membrane anchored proteoglycans such as glypican and/or syndecan family members. We also examined the relationship between the presence of cell layer of syncytiotrophoblasts and the coagulation using clinical specimens, and revealed that the fibrin depositions were specifically observed on the regions lacking syncytiotrophoblasts cell layer in placental villi. Therefore, it is likely that during pregnancy PP5/TFPI-2 may be retained on the surface of placental villi via proteoglycans, and may play an important role to maintain intervillous blood flow and the patency of microvasculature in feto-maternal blood system mediated by the inhibition of serine proteinases involved in the blood coagulation. PMID- 11945081 TI - Hypobaric hypoxia and villous trophoblast: evidence that human pregnancy at high altitude (3600 m) perturbs epithelial turnover and coagulation-fibrinolysis in the intervillous space. AB - Spatial relationships between fibrin-type fibrinoid and regions of villous trophoblast were examined in order to address two main questions: [1] is high altitude pregnancy accompanied by changes in the sizes of trophoblast compartments (cytotrophoblast, syncytiotrophoblast, denudation sites)?, and [2] do highland placentae differ in the amounts and distribution patterns of perivillous fibrin-type fibrinoid? Placentae were collected from two ethnic groups completing term pregnancies at low (400 m above sea level; n=25) and high (3600 m; n=45) altitude in Bolivia. Masson trichrome-stained sections were sampled randomly and analysed stereologically to estimate compartment volumes and surfaces. Comparisons were drawn using variance, Chi-squared and contingency table analyses. At high altitude, birthweights were 265 g lower and placentas had a larger intervillous space (270 cf 181 cm(3)), less fibrin-type fibrinoid (4.1 cf 8.4 cm(3) by volume; 2570 cf 4430 cm(2) by surface area), less villous trophoblast (50 cf 73 cm(3)) and a smaller villous surface (5.6 cf 7.0 m(2)). Volumes were reduced in all syncytiotrophoblast compartments (with and without nuclear aggregations). Cytotrophoblast was maintained and its relative volume increased significantly (from 2.7 to 3.6 per cent of trophoblast volume). Decreases in villous surface area affected primarily thinner (nuclear aggregate free) regions of syncytium. Regardless of altitude, fibrin-type fibrinoid was deposited non-randomly: it was preferentially located at sites of trophoblast denudation. Although no altitudinal differences in fibrin-type fibrinoid patterns were detected, absolute surfaces were diminished on denuded and thinner regions of trophoblast but not on syncytial knots or bridges. Ethnic differences at low altitude (relatively greater deposits on denudations in Amerindians) were minimized at high altitude. We conclude that pregnancy at high altitude alters the epithelial steady state (towards cytotrophoblast and away from syncytiotrophoblast) and the coagulation-fibrinolysis steady state in the intervillous space (to favour fibrinolysis over coagulation). Thinner regions of syncytiotrophoblast may be the main sites of greater fibrinolytic or anticoagulatory activity. The findings are partly consistent with results from in vitro studies which indicate that hypoxia stimulates proliferation of cytotrophoblast but impairs fusion into syncytium. PMID- 11945082 TI - Monoamine oxidase expression and activity in human placentae from pre-eclamptic and normotensive pregnancies. AB - A feature of pre-eclampsia is that circulating levels of maternal serotonin (5 hydroxytryptamine) are elevated and placental monoamine oxidase-A (MAO-A) activity, the major factor in the regulation of serotonin levels in pregnancy, is reduced. It is not known whether this is due to a reduced MAO-A protein content or a reduced catalytic turnover of the serotonin by MAO-A; this question has been addressed in the present work. Term placentae from normotensive and pre-eclamptic women were analysed for MAO-A specific mRNA expression (by semi-quantitative RT PCR), MAO-A protein (by immunohistochemistry and quantitative ELISA, using a MAO A specific monoclonal antibody), together with MAO activity (using [(3)H] labelled 5-hydroxytryptamine as substrate). Immunohistochemical analysis of placentae from both normotensive and pre-eclamptic women demonstrated that MAO-A protein is located in the cytoplasm of the placental syncytiotrophoblast layer, consistent with a mitochondrial location; no MAO-A protein was found in the nucleus. No MAO-B protein was detected in this placental layer, despite the presence of MAO-B mRNA. The results indicate that both total protein/g fresh weight and MAO-A protein/g fresh weight were approximately 40 per cent lower in pre-eclamptic than in normotensive placentae, but that there was no statistical difference in the expression of MAO-A mRNA in relation to GAPDH or actin mRNA or in MAO-A protein/mg total protein. However, MAO-A activity/g fresh weight was significantly reduced in pre-eclamptic placentae, in agreement with previous findings. This was found to be due to a 60 per cent reduction (P< 0.05) in the catalytic turnover (activity/molecule) of the enzyme. This study has therefore clearly shown that the expression of placental MAO-A specific mRNA and MAO-A protein are not specifically affected in pre-eclampsia, but that the catalytic efficiency of the expressed MAO-A enzyme in pre-eclamptic placentae is greatly reduced. PMID- 11945084 TI - Preparation and functional characterization of villous cytotrophoblasts free of syncytial fragments. AB - Recent studies suggest that purified villous cytotrophoblasts are largely contaminated by mononucleated syncytial fragments and therefore unsuitable for studies of trophoblast differentiation. We assessed highly purified (>99.99 per cent) populations of villous trophoblasts for fragment contamination using the syncytial markers placental alkaline phosphatase (PLAP, by immunohistochemistry) and exteriorized phosphatidyl serine (ePS, by flow cytometric analysis). The preparations contained from 4-46 per cent syncytial fragments. However, we find that PLAP negative cells preferentially adhere to tissue culture surfaces and that all preparations were <2 per cent PLAP positive after routine plating and washing procedures. A second purification procedure eliminated dead (propidium iodide permeable) cells and separated viable syncytial fragments (ePS-positive) from viable cytotrophoblasts (ePS-negative) by two colour fluorescence activated cell sorting (FACS). Viable ePS-positive cells were ultrastructurally apoptotic, adhered poorly in culture and those that adhered rapidly underwent apoptosis. Viable ePS-negative cells contained large heterochromic nuclei and cytoplasmic structures, adhered strongly in culture and remained viable. The latter population (putative true villous CT) differentiated into syncytialized cells when cultured with EGF. We conclude that villous CT can be routinely purified, are viable in culture and can undergo syncytial fusion without extensive preformed syncytium. PMID- 11945083 TI - Placental morphogenesis in pregnancies with Down's syndrome might provide a clue to pre-eclampsia. AB - Insufficient perfusion of placenta in pre-eclampsia is commonly associated with oxidative stress leading to increased superoxide formation and reduced invasion of uterine spiral arteries by differentiated migratory cytotrophoblasts. The superoxide dismutase (SOD) level, responsible for eliminating toxic superoxides, drops significantly in pre-eclampsia. On the contrary, the SOD synthesis increases dramatically, compared to that of normal placenta, in pregnancies with trisomy 21 (T21) fetus. However, despite a low level of placental hypoplasia, the overall perfusion of T21 placentae is comparable to that of normal pregnancy. In the light of recent reports on alternative modes of SOD function and factors regulating pathways of cytotrophoblast differentiation, here we have attempted to reconcile the two seemingly disparate pregnancy conditions and suggest that trisomy 21 pregnancies might provide new insight into our understanding of placental morphogenesis in pre-eclampsia. PMID- 11945085 TI - A low conductance, non-selective cation channel from human placenta. AB - Non-selective cation channels have been identified in the plasma membranes of many different cells. Previous research using fluorescent techniques has demonstrated the presence of cation conductances in membranes from human trophoblast. The purpose of this work was to explore, by electrophysiological methods, a non-selective cation channel in apical membranes from human placenta. Human placental apical membranes were purified by differential centrifugation and reconstituted in giant liposomes. These giant liposomes were then used for electrophysiological studies and were probed for the presence of cation channels by the patch-clamp method. The channel identified had a linear current-potential relationship with a conductance of around 16 pS in symmetrical Na(+) solution. Under asymmetrical conditions the reversal potential was close to the reversal potential for Na(+). The channel was equally permeable to sodium and potassium and the permeability sequence was NH+4>Cs(+) approximately Rb(+)>Na(+) approximately K(+)>Li(+). The channel also showed permeability to calcium and barium. The channel was insensitive to calcium but was blocked by millimolar concentration of Mg(2+). We have demonstrated the presence of a low conductance, non-selective cation channel in placental apical membranes. These channels share some properties with non-selective cation channels previously described in other different cells. The precise role of these channels in placental physiology has yet to be determined. PMID- 11945086 TI - Localization of the extracellular Ca(2+)-sensing receptor in the human placenta. AB - We have demonstrated using immunohistochemistry and in situ hybridization that the calcium-sensing receptor (CaR) is expressed in both villous and extravillous regions of the human placenta. CaR expression was detected in both first trimester and term placentas. In the villous region of the placenta, the CaR was detected in syncytiotrophoblasts and at lower levels in cytotrophoblasts. Local expression of the CaR in the brush border of syncytiotrophoblasts suggests a role for maternal Ca(2+) concentration in the control of transepithelial transport between the mother and fetus. In the extravillous region of the placenta, the CaR was detected in cells forming trophoblast columns in anchoring villi, in close proximity to maternal blood vessels and in transitional cytotrophoblasts. Given the importance of extravillous cytotrophoblasts in the process of uterine invasion and maintenance of placental immune privilege, the CaR represents a possible target by which the maternal extracellular Ca(2+) concentration could promote or maintain placentation. Thus, the results support hypotheses that the CaR contributes to the local control of transplacental calcium transport and to the regulation of placental development. PMID- 11945087 TI - Monochorionic twins and twin-twin transfusion syndrome: the protective role of arterio-arterial anastomoses. AB - Unidirectional arterio-venous (AV) anastomoses often result in twin-twin transfusion syndrome (TTTS). Additional oppositely directed anastomoses may compensate for the circulatory imbalance and either prevent, delay the onset, or moderate the severity of TTTS. Intuitively, higher pressure gradient, oppositely directed AV anastomoses (indicated as VA) would be expected to compensate better for TTTS than lower pressure gradient arterio-arterial (AA) anastomoses. However, clinical evidence suggests AA anastomoses compensate more efficaciously, because virtually all non-TTTS monochorionic twin placentas have AAs (84 per cent), contrary to TTTS placentas, where only 30 per cent have an AA. We sought to explain this observation by comparing the capabilities of various size VA and AA anastomoses to compensate for the effects of the primary AV. As study design we used a previously developed mathematical computer model of TTTS to determine ranges of anastomotic vascular resistances which cause varying fetal and amniotic fluid discordances. Anastomotic resistances were related with the radii of their feeding vessels, using fractal geometry modelling to mimic the placental vascular tree, and various assumptions regarding arterial blood flow. The results were as follows. An AA anastomosis of equal size as the feeding artery of an AV or VA has a significantly smaller resistance. The primary AV anastomosis may be compensated by both VA as well as AA anastomoses. However, VA transfusion adequately compensates AV flow only for a small range of VA to AV vascular radius ratios. In contrast, AA transfusion compensates the AV flow for a much wider range of AA to AV vascular radius ratios. In conclusion, the wider range of AA than VA radii for adequate compensation of the AV explains the finding that an AA protects more frequently than a VA of similar size against the manifestations of TTTS. These results may possibly allow future risk stratification of monochorionic twins by non-invasive sonographic assessment of the size and type of anastomoses. PMID- 11945088 TI - Placenta accreta--summary of 10 years: a survey of 310 cases. AB - The objective was to study the incidence, risk factors, and outcome of pregnancies complicated by placenta accreta in our population. Retrospective analysis of all deliveries between the years 1990-2000, and identification of all cases of placenta accreta, defined by clinical or histological criteria. For comparison purposes we defined two sub-groups: (i) all cases that ended with severe outcome and (ii) all patients who had a previous event of placenta accreta in one or more of their previous deliveries. We evaluated the potential risk factors leading to these conditions. The SPSS software package was used for statistical analysis. Univariate and multivariate analyses were performed by stepwise logistic regression. The study covered 34 450 deliveries from which 310 cases of placenta accreta were diagnosed (0.9 per cent). The risk factors associated with placenta accreta were previous cesarean delivery (12 per cent), advanced maternal age, high gravidity, multiparity, previous curettage and placenta previa (10 per cent). Hysterectomy was performed in 11 patients (3.5 per cent) with one case of maternal death, whereas 21 per cent of the patients required postpartum blood products transfusion. Antenatal diagnosis of placenta accreta or percreta by ultrasound or MRI, was achieved only in eight of the cases. In the sub-group of 15 patients (4.8 per cent) with severe outcome, the only significant risk factors were increased parity (O.R.=1.29, 95 per cent CI 1.056-1.585), anteriorly low placenta (O.R.=6.1, 95 per cent CI 1.4-25.3) and repeated cases of caesarean sections (O.R.=3.3, 95 per cent CI 0.9-12.5), whereas in the 49 (16 per cent) patients with repeated cases of placenta accreta the only significant risk factor was the number of deliveries (O.R.=1.5, 95 per cent CI 1.0-2.2). Repeated cesarean delivery, high parity, and anteriorly low placental location are associated with severe outcome in case of placenta accreta. Women with repeated events of placenta accreta may have better outcome and a genetic factor may serve as a cause for this condition. PMID- 11945089 TI - Expression of apoptosis in placentae from mice lacking the prostaglandin F receptor. AB - This study aimed to investigate the changes in apoptosis in the placenta and decidua of pregnant mice lacking the prostaglandin F receptor. Mouse placentae were removed from fetuses on days 10-23 of pregnancy. Apoptotic cells were examined by a DNA fragmentation assay and the terminal deoxynucleotidyl transferase-mediated dUDP nick end-labelling (TUNEL) technique. The placenta and decidual weight increased before day 18 and 14 of pregnancy, and then decreased with gestational day. After day 19, the fetuses gradually died in the uterus. All fetuses died in the uterus on day 23 of pregnancy. The number of apoptosis was not significantly different between wild type and FP-deficient mice before day 18 of pregnancy by DNA fragmentation and TUNEL staining. The DNA fragmentation was always more pronounced in decidual tissue on each day of pregnancy. DNA laddering on placentae was more extensive on day 22 than day 18. In placenta, most TUNEL positive cells were detected in trophoblast and stromal cells. A higher intensity of apoptotic cells was in the decidual basalis. The main area was the centre of the decidual basalis, and was in decrease toward to margin of placenta. The index of TUNEL positive cells increased as gestation progressed toward termination. Especially, it was prominent in the placentae on day 22 compared with that day 18 of pregnancy. The increased TUNEL-positive staining in syncytiotrophoblast surface was found in placenta at post-term, compared with those at term. Apoptosis may provide insights into both normal placental development and placental dysfunction during an abnormal pregnancy from post-term pregnancy. PMID- 11945090 TI - Increased apoptosis of human fetal membranes in rupture of the membranes and chorioamnionitis. AB - Apoptosis is thought to participate pathophysiologically in the rupture of human fetal membranes (ROM). The aim of this study was to assess apoptosis of the amnion and the chorion in relation to ROM and chorioamnionitis (CAM). The amnion and chorion at the position of the cervical os and fundus of the uterus were obtained from 44 patients. Apoptotic DNA fragmentation was densitometrically determined, and the relative ratio was used for the quantitative evaluation. Among patients without CAM, the relative ratios of apoptosis in the amnion from patients with ROM were higher than those in patients without ROM (P< 0.05). Among patients without ROM, the apoptotic levels in the amnion from patients with CAM were higher than those in patients without CAM (P< 0.05). These were the cases with the amnion at the position of cervical os and fundus, but not with the chorion. The highest ratio of apoptosis was seen in the amnion from patients with CAM and ROM. Among patients with ROM and no CAM, the apoptotic levels at the cervical os in the amnion (P=0.059) and chorion (P< 0.05) was higher than those at the fundus. The increased apoptosis of human fetal membranes was related to ROM and CAM. Apoptosis plays a role in the pathophysiology of ROM. PMID- 11945091 TI - Trophoblast invasion of spiral arteries: a novel in vitro model. AB - Extravillous trophoblasts invade the uterine wall (interstitial invasion) and the spiral arteries (endovascular invasion), replacing the cells of the vessel wall and creating a high-flow low-resistance vessel. We have developed a novel model to allow the interactions between the invading trophoblast cells and the cells of the spiral artery to be directly examined. Unmodified (non-placental bed) spiral arteries were obtained from uterine biopsies at caesarean section. Fluorescently labelled trophoblasts were seeded on top of artery segments embedded in fibrin gels (to study interstitial invasion) or perfused into the lumen of arteries mounted on a pressure myograph (to study endovascular invasion). Trophoblasts were incubated with the vessels for 3-5 days prior to cryo-sectioning. Both interstitial and endovascular interactions/invasion could clearly be detected and a comparison of the extravillous trophoblast cell line, SGHPL-4 and primary first trimester cytotrophoblasts showed both to be invasive in this model. This novel method will prove useful in an area where in vitro studies have been hampered by the lack of suitable models directly examining cellular interactions during invasion. PMID- 11945092 TI - Formation of monozygotic twins: when does it occur? AB - An individual embryo from day 15 of gestation in the rhesus monkey showed partial separation of the cranial end of the embryonic shield into two separate cell masses with separate amnions but with a common yolk sac. Although what the fate of this conceptus might have been is not known, it demonstrates that the physical features of implantation in this primate are permissive of relatively late division of the epiblast, which suggests that monozygotic twinning may occur as late as the embryonic shield stage just prior to primitive streak formation. PMID- 11945093 TI - Refractory epilepsy: a progressive, intractable but preventable condition? AB - Intractable seizures are just one manifestation of 'refractory epilepsy', which can be recognized as a distinct condition with multifaceted dimensions, including neurobiochemical plastic changes, cognitive decline and psychosocial dysfunction, leading to dependent behaviour and a restricted lifestyle. The biological basis of 'refractoriness' is likely to be multifactorial, and may include the severity of the syndrome and/or underlying neuropathology, abnormal reorganization of neuronal circuitry, alteration in neurotransmitter receptors, ion channelopathies, reactive autoimmunity, and impaired antiepileptic drug (AED) penetration to the seizure focus. Some of these deleterious changes may be a consequence of recurrent seizures. We hypothesize that 'refractory epilepsy' may be prevented by interrupting this self-perpetuating progression. There is increasing evidence that these patients can be identified early in the clinical course and, thus, be targeted early for effective therapeutic intervention. Failure of two first-line AEDs due to lack of efficacy or poor tolerability should prompt consideration of epilepsy surgery in a patient with a resectable brain abnormality. For the majority not suitable for 'curative' surgery, AEDs should be combined with the aim of achieving 'synergism'. This strategy has the potential to improve outcome by preventing the insidious progression to intractable 'refractoriness' and a downward spiraling quality of life. PMID- 11945094 TI - Improving diagnostic procedure and treatment in patients with non-epileptic seizures (NES). AB - The purpose of this study was to examine the incidence of patients with NES vs. NES and concomitant epilepsy in an epilepsy centre and to present a diagnostic algorithm. We collected and reviewed the data of 322 patients consecutively referred to the adult ward of our epilepsy centre in 1 year. The results of our study reveal that 44 (14%) of all patients referred had NES. Of these, nine proved to have concomitant epilepsy. Of 44 patients with NES, 20 were treated with AED on admission. In 14 cases this unnecessary antiepileptic drug treatment was stopped. In six remaining patients with NES and concomitant epilepsy, the total number of AEDs could be reduced until discharge. The maximum duration of AED treatment among patients with NES only, had been longer than 360 months (median 72 months). Much has been written about whether the diagnosis of psychogenic non-epileptic events is overused. According to our experience however, the fact that many patients with so-called 'pharmacoresistant epilepsy', suspected NES or other diagnoses are referred to a centre of excellence much too late, proves to be the key problem in diagnosis and treatment of NES. We conclude that early admission of so-called 'pharmacoresistant epilepsy' to an epilepsy centre, establishing a standard work-up and clarifying the medical terminology will improve diagnosis and lead to adequate therapy of NES as well as prevent unnecessary drug treatment. PMID- 11945095 TI - Systematic reviews of specialist epilepsy services. AB - BACKGROUND: Concern has been expressed over UK epilepsy service standards but the most clinically effective model of care is unknown. OBJECTIVE: To systematically review the current evidence on specialist epilepsy clinics compared to general neurology clinics and specialist epilepsy nurses compared to usual care. METHODS: Medline, Psychlit, Embase, Healthplan, GEARS, BIDS ISI, UKCHHO, international HTA websites, InterTASC databases and The Cochrane Library were searched to September 1999. Any studies comparing specialist epilepsy clinics or nurses to generalist services or usual care, reporting physical health, costs or generic quality-of life outcomes were included. Two people independently applied inclusion and exclusion criteria and extracted data independently. Randomized controlled trial (RCT) quality was assessed by Jadad score and other studies qualitatively by the likelihood of bias. RESULTS: Findings were one RCT and two other studies on epilepsy clinics and four RCTs and a controlled trial on epilepsy nurses. Data synthesis was inappropriate. Epilepsy clinics showed no evidence of reduced seizure frequency or severity, no quality-of-life information and were more expensive. Epilepsy nurse services showed no evidence of reduced seizure frequency or severity, no effect on quality-of-life but were less expensive. CONCLUSION: There is insufficient evidence to demonstrate the superiority of any particular care model for producing better health outcomes. PMID- 11945096 TI - Assessment of the utility of paediatric electroencephalography. AB - Electroencephalography (EEG) is an important tool in investigating children with neurological disorders, particularly epilepsy. The objectives were to examine the relationship between clinical indications and EEG results, and assess the predictability of a normal result. 438 consecutive paediatric EEGs were included prospectively. One certified electroencephalographer (EEGer) reviewed EEG requisitions and recorded his prediction of a normal result. EEGs were reviewed separately and the relationship between the clinical indications and EEG abnormalities was recorded. The children's mean age was 5 years (SD 4.2). Paediatric neurologists ordered 32% of EEGs. The first EEG was studied in 65% of cases. Overall, 55% of the EEGs were abnormal. Repeat EEGs were twice as likely to be abnormal (95% CI 1.3-3, P= 0.001). Established epilepsy, using antiepileptic drugs, and sleep record, highly correlated with an abnormal result ( P< 0.0001). The EEGer predicted 26% of the EEGs to be normal. A normal EEG was correctly predicted in 97% of non-epileptic paroxysmal events, however, normalization of EEG was correctly predicted in only 54% of children with seizures. EEGs of 15 (3.4%) children with epilepsy revealed unexpected findings that completely changed their management. To conclude, a normal EEG is highly predictable in non-epileptic paroxysmal events. EEGs of children with epilepsy are not predictable and may yield unexpected results. PMID- 11945097 TI - Remacemide hydrochloride as an add-on therapy in epilepsy: a randomized, placebo controlled trial of three dose levels (300, 600 and 800 mg/day) in a B.I.D. regimen. AB - Remacemide hydrochloride is a low-affinity, non-competitive NMDA receptor channel blocker under investigation for the treatment of epilepsy. This double-blind, placebo-controlled, multicentre study assessed the safety and efficacy of adjunctive remacemide hydrochloride or placebo, in adult patients with refractory epilepsy who were already taking up to three antiepileptic drugs (including an enzyme-inducer). Patients (n= 262) were randomized to one of three doses of remacemide hydrochloride (300, 600 or 800 mg/day) or placebo, in a B.I.D. regimen, for up to 14 weeks. Plasma concentrations of carbamazepine (CBZ) and phenytoin (PHT) were controlled throughout. Patients recorded their seizures on a diary card. There was an increase in the percentage of responders (defined as a reduction in seizure frequency from baseline > or = 50 %), from 15 % (9/60) with placebo, to 30 % (18/60) in the 800 mg/day group. A pairwise comparison between remacemide hydrochloride 800 mg/day and placebo was statistically significant (P = 0.049). Most reported adverse events (mainly CNS and gastrointestinal) were mild or moderate in severity and dose-dependent. Adjunctive remacemide hydrochloride treatment was associated with a higher, dose-related responder rate compared with placebo. The difference reached significance at the highest dose tested (800 mg/day). Remacemide hydrochloride was well tolerated. PMID- 11945099 TI - Coma probably induced by lorazepam-valproate interaction. AB - Both valproate (VPA) and lorazepam (LZP) are primarily cleared from the body by glucuronidation. Concomitant administration of VPA has been reported to reduce the elimination of LZP. However, it remains unknown whether this drug interaction is clinically significant. We report a patient with epilepsy who showed that VPA LZP interaction could result in severe encephalopathy such as coma. PMID- 11945098 TI - Remacemide hydrochloride as an add-on therapy in epilepsy: a randomized, placebo controlled trial of three dose levels (300, 600 and 1200 mg/day) in a Q.I.D. regimen. AB - Remacemide hydrochloride is a low-affinity, non-competitive N-methyl-D-aspartic acid (NMDA) receptor channel blocker, under investigation in epilepsy. This double-blind, placebo-controlled, multicentre study assessed the safety and efficacy of remacemide hydrochloride or placebo, as adjunctive therapy, in 252 adult patients with refractory epilepsy who were already taking up to three antiepileptic drugs (including an enzyme-inducer). Patients were randomized to one of three doses of remacemide hydrochloride (300, 600 or 1200 mg /day) or placebo Q.I.D., for up to 15 weeks. An increasing percentage of responders (defined as a reduction in seizure frequency from baseline of > or =50%) was seen with increasing remacemide hydrochloride dose. At 1200 mg /day, 23% of patients were responders compared with 7% on placebo. This difference was significant (P = 0.016), as was the overall difference between treatments (P = 0.038). Adverse events: dizziness, abnormal gait, gastrointestinal disturbance, somnolence, diplopia and fatigue were mild or moderate in severity. Carbamazepine and phenytoin plasma concentrations were well controlled and maintained within target ranges, with no evidence of improved seizure control due to increases in the concentrations of these drugs. A dose-dependent, significant, increase in responders following adjunctive remacemide hydrochloride compared with placebo was observed. Remacemide hydrochloride was well tolerated. PMID- 11945100 TI - The long-term course of seizure susceptibility in two patients with juvenile myoclonic epilepsy. AB - We have observed epileptic seizures of juvenile myoclonic epilepsy (JME) to be surprisingly sensitive to higher mental activity. The purpose of the present study was to examine changes over time in seizure susceptibility in two patients with JME who we followed-up for over 20 years. During the period, they were repeatedly subjected to provocative cognitive tasking, that is, to 'neuropsychological EEG activation'. Tasks included reading, speaking, writing, written arithmetic, mental calculation, and spatial construction. During the first 15 years after the onset of symptoms, higher mental activities, mainly associated with use of the hands, i.e. writing, written calculation, and spatial construction, as well as physiological factors, such as sleep deprivation, awakening, and fatigue, precipitated the seizures. Generalized tonic-clonic and absence seizures but not myoclonic seizures disappeared almost completely after antiepileptic treatment. After age 30, the provocative effect of higher mental activities persisted, and the myoclonic seizures decreased under same drug regimen. These observations suggest that the pathophysiology of JME improves with time but persists for a long time, and that it is closely related to a neural network involved in higher mental activities mainly associated with use of the hands rather than in physiological factors emphasized in prior reports. PMID- 11945102 TI - Palatal myoclonus responding to lamotrigine. AB - Palatal myoclonus (PM) is a rare neurological condition with no established treatment. We report on a case of PM with a good response to lamotrigine. PMID- 11945101 TI - Temporal lobe epilepsy of adult age of possible idiopathic nature. AB - OBJECTIVE: To verify the existence of sporadic cases of temporal lobe epilepsy (TLE) of possible idiopathic nature with onset in adult life and to define their characteristics. BACKGROUND: Currently, the existence of these forms is not fully recognized. In the last few years, familial cases of TLE of the early adult age have been described. Besides, sporadic cases of TLE characterized by both onset in adulthood and high prevalence of familial history of epilepsy have also been reported. METHOD: All subjects with TLE, defined according to the ILAE criteria (1989), with onset in adult age, followed at our Centre from 1990 to 1998, were re-examined to find out the cases with possible idiopathic origin of seizures. RESULTS: Among 159 subjects with a diagnosis of TLE, 14 subjects (8.8%) were so identified (Group 1), and compared with the remaining 145 subjects (Group 2). High familial incidence for epilepsy, prevalence of autonomic seizures, low seizure frequency, and good prognosis are the main characteristics of the group 1. The difference between the two groups is statistically significant for family history for epilepsy (chi square test: P< 0.01) and for freedom from seizures after medication (chi square test: P< 0.05). CONCLUSION: The distinctive features of the subjects identified suggest that at least some of these patients may have familial TLE. Whether this disorder may be present in sporadic form remains a matter of debate which must await the identification of the gene(s) responsible of this disorder. Till then, it is worthwhile to underline the better prognosis of these patients compared to those that did not meet these criteria. PMID- 11945103 TI - Epileptic seizures, some dyskinesia and the 'alpha tape'. PMID- 11945104 TI - Visual field defects. PMID- 11945105 TI - Optimal timing of thrombolytic therapy in acute ischaemic stroke. AB - The clinical benefit of thrombolytic therapy for patients experiencing acute cerebral ischaemia has been demonstrated by both clinical trials and phase IV studies. However, such treatments must be initiated in a rapid manner, with treating physicians adhering to strict protocols designed to minimise delays and maximise safety. The efficacy of intravenous drug administration has been established with alteplase (recombinant tissue plasminogen activator; tPA) and ancrod, but only if these drugs can be administered within 3 hours of symptom onset. The use of alteplase beyond this timeframe, or outside of established protocols, may be hazardous. The use of alternative intravenous thrombolytic agents, such as streptokinase, also appears hazardous. Intra-arterial delivery of thrombolytic drugs such as pro-urokinase may extend clinical benefit to the 6 hour time frame. PMID- 11945106 TI - Pharmacological treatment of psychotic agitation. AB - The presentation of agitated psychotic patients to psychiatric emergency services is a common occurrence. The traditionally accepted treatment for such patients involves the use of a typical antipsychotic, generally haloperidol. More recently benzodiazepines, such as lorazepam, have been used in combination with antipsychotics due to their sedative properties and relatively benign adverse effect profiles. Standard clinical protocol at many institutions involves the intramuscular administration of 5 to 10mg of haloperidol and 1 to 2mg of lorazepam. Atypical antipsychotics have gained acceptance as first-line treatments for psychotic disorders. These drugs are seen as an improvement over traditional antipsychotics because of their increased efficacy and reduced extrapyramidal effects. The utility of atypical antipsychotics in the emergency setting has been relatively unexplored because slow titration schedules or dose limiting adverse effects for some members of the class have made this form of treatment impractical. However, the recent availability of oral liquid and rapidly dissolving tablet preparations of some atypical agents has provided useful alternatives in some cases. Nevertheless, for many patients a parenteral drug is the only desirable or feasible treatment option. Intramuscular preparations of the atypical antipsychotics olanzapine and ziprasidone have been developed, and are close to launch in the US. The availability of a rapid-acting intramuscular preparation of an atypical antipsychotic could represent a significant advancement in the treatment of agitation associated with psychosis. PMID- 11945108 TI - Advances in atypical antipsychotics for the treatment of schizophrenia: new formulations and new agents. AB - Innovation in atypical antipsychotic agents continues with new preparations of available drugs as well as novel agents. In this article, we provide an update on these novel products by reviewing information from a computerised literature search, recent abstracts and discussions with industry representatives. A generic formulation of clozapine is now available. It may be less well absorbed and/or less effective than Clozaril, although evidence is conflicting. A fatty acid amide derivative of clozapine is in early development. A liquid formulation of risperidone is currently available, which may be a useful treatment for psychotic agitation as well as a preferable alternative to tablets for some patients. A depot formulation is in development for the long-term management of psychosis. An orally disintegrating tablet formulation of olanzepine is a useful alternative to standard tablets. A short-acting injectable formulation of the drug is in development for psychotic agitation. Sachets and slow-release formulations of quetiapine are in development. Ziprasidone, a recently launched agent, is available in tablet form for schizophrenia/schizoaffective disorder, psychotic depression and mania. A short-acting injectable formulation is in development for psychotic agitation. Aripiprazole (tablets) and iloperidone (tablets and depot injection) are two antipsychotics in development for schizophrenia/schizoaffective disorder (available information regarding iloperidone is very limited). These new formulations and agents should broaden options for the treatment of psychosis. PMID- 11945109 TI - Interactions between antiepileptic drugs and hormonal contraception. AB - An interaction between antiepileptic drugs (AEDs) and the combined oral contraceptive pill was first proposed when the dose of estradiol in the oral contraceptive pill was reduced from 100 to 50 microg. There was a higher incidence of breakthrough bleeding and contraceptive failure among women with epilepsy compared with women in general. Since then, interaction studies have been undertaken to look for possible interactions between AEDs and the combined oral contraceptive pill. Phenobarbital (phenobarbitone), phenytoin, carbamazepine, oxcarbazepine, felbamate and topiramate have been shown to increase the metabolism of ethinylestradiol and progestogens. Therefore, if a women is on one of the AEDs and wishes to take the oral contraceptive pill, she will need to take a preparation containing at least 50 microg of ethinylestradiol. Levonorgestrel implants are contraindicated in women receiving these AEDs because of cases of contraceptive failure. It is recommended that medroxyprogesterone injections be given every 10 rather than 12 weeks to women who are receiving AEDs that induce hepatic microsomal enzymes. There are no interactions between the combined oral contraceptive pill, progesterone-only pill, medroxyprogesterone injections or levonorgestrel implants and the AEDs valproic acid (sodium valproate), vigabatrin, lamotrigine, gabapentin, tiagabine, levetiracetam, zonisamide, ethosuximide and the benzodiazepines. Therefore, normal dose contraceptive preparations can be used in patients receiving these AEDs. PMID- 11945111 TI - Monitoring the safety of vaccines: assessing the risks. AB - The safety of vaccines, particularly the most widely used vaccines to which most children are exposed as infants and toddlers, has always been an extremely high priority for vaccine manufacturers and government agencies. Products intended for healthy people must be held to a high standard of safety assurance. In addition to the intense safety assessments conducted prior to licensure, post-marketing surveillance programmes are essential to identify and study possible risks that occur too rarely to have been identified in pre-licensure studies or that occur in populations not studied in pre-licensure studies. Studying rare risks of vaccines is more complex than for therapeutic products because the exposure is virtually universal for many vaccines, ensuring occurrence simply by chance of many adverse outcomes in temporal association with vaccination. In the US the Vaccine Safety Datalink (VSD), a consortium of managed care organisations, has been established to study more rigourously possible vaccine-associated risks. These risks may be identified through reports to the Vaccine Adverse Event Reporting System (VAERS), the nationwide passive surveillance programme, as well as other sources. The combination of passive surveillance and more structured case-control or cohort studies possible in the VSD has helped to both identify new vaccine risks and to provide reassuring evidence of lack of risk in other situations where concerns have been raised. PMID- 11945107 TI - Interactions between psychotropics, anaesthetics and electroconvulsive therapy: implications for drug choice and patient management. AB - Despite many predictions that electroconvulsive therapy (ECT) would be replaced by pharmacotherapy, ECT has remained an invaluable adjunct in the management of severe psychiatric disease. Both pharmacotherapy and ECT continue to be used extensively, and will frequently be administered concurrently. The majority of patients requiring ECT will need anaesthesia; therefore, interactions could conceivably occur between the psychotropic drugs, ECT and the anaesthetic agents utilised. In managing an anaesthetic for ECT the effects of the anaesthetic agents and other medications on seizure intensity are important determinants influencing outcome. With regard to the antidepressants, tricyclic antidepressants (TCAs) and ECT can be combined safely and beneficially. More care is required when ECT is administered in the setting of a monoamine oxidase inhibitor (MAOI), especially the older irreversible varieties and in patients recently placed on MAOI therapy. Of the anticonvulsants and mood stabilisers, lithium and ECT given concurrently add significant risk of delirium and/or organic syndromes developing. Possible concerns with valproate, carbamazepine, lamotrigine, gabapentin and topiramate are that they may inhibit seizure activity. Additionally, carbamazepine may prolong the action of suxamethonium (succinylcholine). The combination of antipsychotics and ECT is well tolerated, and may in fact be beneficial. As regards the anxiolytics, benzodiazepines have anticonvulsant properties that might interfere with the therapeutic efficacy of ECT. CNS stimulants on the other hand may prolong seizures as well as produce dysrhythmias and elevate blood pressure. Calcium channel antagonists should be used with great care to avoid significant cardiovascular depression. The anaesthesiologist should therefore remain vigilant at all times, as untoward responses during ECT might occur suddenly due to interactions between psychotropics, anaesthetic agents and/or ECT. PMID- 11945110 TI - Discontinuation syndrome in dysthymic patients treated with selective serotonin reuptake inhibitors: a clinical investigation. AB - OBJECTIVE: Many authors have reported discontinuation symptoms associated with selective serotonin reuptake inhibitors (SSRIs). The aim of this study was to investigate the incidence and characteristics of the discontinuation syndrome in patients who stopped treatment with the SSRIs paroxetine and fluoxetine under the usual conditions of clinical practice, and to identify clinical predictors of the syndrome. METHODS: Ninety-seven outpatients who received an initial diagnosis of dysthymic disorder, who responded to >or=8 weeks treatment with paroxetine (n = 52) or fluoxetine (n = 45), and who discontinued the SSRI according to their psychiatrist's instructions were included. They were assessed at the time of discontinuation using a semi-structured interview for clinical and treatment characteristics, the Hamilton Depression Rating Scale (HAM-D) and the Montgomery Asberg Depression Rating Scale (MADRS). Patients were then assessed 4 weeks later using a checklist for discontinuation symptoms, a semi-structured interview for discontinuation symptom characteristics, and the HAM-D and the MADRS. RESULTS: A discontinuation syndrome was found in 26 patients (26.8% of our sample); of this group, 22 patients (84.6%) had received paroxetine, and 4 patients (15.4%) had received fluoxetine. The mean time at onset of symptoms was 2 days after drug discontinuation and the mean duration was 5 days. The statistical comparison between the groups with and without a discontinuation syndrome found two significant differences - a discontinuation syndrome was more common in patients treated with paroxetine and in patients with an earlier onset of dysthymic disorder. Multiple regression analysis confirmed that these two factors were related to the duration of discontinuation symptoms, while the number of symptoms was associated with three factors, including use of paroxetine, age at onset of dysthmia and female gender. CONCLUSIONS: A discontinuation syndrome is common after treatment with SSRIs is stopped in patients with dysthymia, and it appears to be more common in patients receiving paroxetine than in those receiving fluoxetine. The syndrome is related both to drug and clinical characteristics. The features of the syndrome in patients with different Axis I diagnoses should be compared in further investigations. PMID- 11945112 TI - Cardiac and CNS toxicity of levobupivacaine: strengths of evidence for advantage over bupivacaine. AB - Bupivacaine is currently the most widely used long-acting local anaesthetic. Its uses include surgery and obstetrics; however, it has been associated with potentially fatal cardiotoxicity, particularly when given intravascularly by accident. Levobupivacaine, a single enantiomer of bupivacaine, has recently been introduced as a new long-acting local anaesthetic with a potentially reduced toxicity compared with bupivacaine. Numerous preclinical and clinical studies have compared levobupivacaine with bupivacaine and in most but not all studies there is evidence that levobupivacaine is less toxic. Advantages for levobupivacaine are seen on cardiac sodium and potassium channels, on isolated animal hearts and in whole animals, anaesthetised or awake. In particular the intravascular dose of levobupivacaine required to cause lethality in animals is consistently higher compared with bupivacaine. In awake sheep, for example, almost 78% more levobupivacaine was required to cause death. In contrast, in anaesthetised dogs no differences were seen in the incidence of spontaneous or electrical stimulation- induced ventricular tachycardia and fibrillations among animals exposed to levobupivacaine or bupivacaine. The reversibility of levobupivacaine-induced cardiotoxicity has also been assessed. Some data point to an advantage of levobupivacaine over bupivacaine but this potential advantage was not confirmed in a recent study in anaesthetised dogs. Three clinical studies have been conducted using surrogate markers of both cardiac and CNS toxicity. In these studies levobupivacaine or bupivacaine were given by intravascular injection to healthy volunteers. Levobupivacaine was found to cause smaller changes in indices of cardiac contractility and the QTc interval of the electrocardiogram and also to have less depressant effect on the electroencephalogram. Assuming that levobupivacaine has the same local anaesthetic potency as bupivacaine, then, all things being equal, it is difficult to argue that levobupivacaine should not displace bupivacaine as the long-acting local anaesthetic of choice. It would appear, however, that levobupivacaine has not yet significantly displaced bupivacaine from the markets in which it is sold. This may be due to a lack of perceived safety benefit and/or consideration of the additional costs that are associated with switching to levobupivacaine, which is approximately 57% more expensive than bupivacaine. If the price of levobupivacaine were closer to bupivacaine then the argument to switch to levobupivacaine would undoubtedly be much stronger. With the continued clinical use of levobupivacaine the database available to make comparisons will increase and this may allow cost-benefit arguments to be made more forcefully for levobupivacaine in the future. PMID- 11945113 TI - Cyclo-oxygenase-2 inhibitors and the kidney: a case for caution. AB - Cyclo-oxygenase (COX) is one of the key enzymes in the biosynthesis of prostaglandins. Two isoforms of this enzyme COX-1 and COX-2 are known to exist. Among other functions, prostaglandins play an important role in the protection of the gastric mucosa and maintenance of renal function in pathophysiological conditions which would otherwise threaten it. Conventional nonsteroidal anti inflammatory drugs (NSAIDs) block prostaglandin synthesis, resulting in gastric mucosal injury and renal dysfunction in susceptible individuals. The recent introduction of selective COX-2 inhibitors, celecoxib and rofecoxib, appear to induce less gastrointestinal morbidity. Although conclusive data are still lacking, there is evidence to suggest that COX-2 antagonists may be capable of causing some of the same renal syndromes seen in association with the older, less selective NSAIDs. PMID- 11945115 TI - Growth hormone therapy and its relationship to insulin resistance, glucose intolerance and diabetes mellitus: a review of recent evidence. AB - It is widely recommended that consideration should be given to the therapeutic use of growth hormone (GH) in adults with GH deficiency, whether the condition is of childhood or adult onset. One reason for this recommendation is the possibility that such treatment may reduce the excess cardiovascular risk which is associated with hypopituitarism. This excess risk has been well documented, with mortality ratios of 1.7 to 2.2 being quoted in different studies, and may be a result of the insulin resistance which occurs in hypopituitarism. However, it has also been suggested that this insulin resistance may itself be the result of GH deficiency, especially as GH deficiency is accompanied by suggestive morphological features such as central adiposity. There is, however, no direct evidence that the increase in cardiovascular risk in hypopituitarism is the result of GH deficiency, and the only prospective study designed to examine the relationship failed to find a statistically significant correlation between the two. Since GH administration may also have an independent adverse effect on insulin sensitivity and could thus cause a theoretical worsening of cardiovascular risk, it is important to review the observed effects of GH administration on carbohydrate metabolism in practice. Interpretation of the literature is made difficult by many confounding factors, including differences in study duration, biochemical tools adopted, the use of selected populations and the dose-dependent effect of GH on synthesis of insulin- like growth factor-1. One of the most sensitive markers of a deterioration in insulin sensitivity is the serum insulin level. A rise in serum insulin (fasting, or post-glucose load) was reported in all studies in which it was measured. The majority of studies have also reported a rise in fasting blood glucose. A smaller proportion of reports noted an associated increase in postprandial glucose and in glycosylated haemoglobin (HbA(1c)) while a few reported new cases of either impaired glucose tolerance or frank diabetes mellitus. In general, however, the observed deterioration in insulin sensitivity was small and increases which occurred in blood glucose were small. Nevertheless, these data indicate that rather than lead to an improvement in insulin resistance in hypopituitarism, GH treatment may actually make it worse. As it is also known that even minor reductions in insulin sensitivity may be associated with a clinically significant increase in cardiovascular risk, further large-scale controlled trials are required before the efficacy and safety of GH treatment of adults can be established. PMID- 11945116 TI - A post-marketing surveillance study of formoterol (Foradil): its use in general practice in England. AB - OBJECTIVE: At the time of marketing, experience of long-term use of prescription medicines in general clinical practice is limited. Postmarketing surveillance is particularly important at this time when medicines may be prescribed to large numbers of patients of all ages, for long-term use. Following marketing of formoterol (Foradil) in the UK in 1996 we undertook a postmarketing surveillance study of formoterol use in general practice. DESIGN: A non-interventional observational cohort study was conducted using the technique of prescription event monitoring. Exposure data were obtained from prescription details; outcome data from questionnaires sent to general practitioners approximately 12 months after the first prescription was dispensed for individual patients. Incidence rates were calculated for reported events, reasons for stopping treatment and outcomes of pregnancy were determined. RESULTS: Data were collected for 5777 patients aged 3 to 96 years, 65% of whom continued treatment for >12 months. Formoterol was prescribed 'off label', to 258 children, (4.5%) of the cohort. The most commonly reported events excluding those related to respiratory disease, were headache, tremor, palpitation, cramp and nausea/vomiting. These events were also among the more common reasons for stopping treatment and reported as suspected adverse drug reactions. 33 patients took formoterol during pregnancy. The cause of death was established for 186 of the 190 deaths (3% of cohort). CONCLUSIONS: Formoterol appears to have been well tolerated by the majority of patients in this study. The most frequently reported adverse events were those known to be associated with beta(2)-agonists, although the frequency of nausea/ vomiting was greater than given in Summary of Product Characteristics. PMID- 11945114 TI - Safety and efficacy of disease-modifying anti-rheumatic agents: focus on the benefits and risks of etanercept. AB - The traditional approach to the treatment of rheumatoid arthritis (RA) has been the use of nonsteroidal anti-inflammatory drugs usually in combination with a disease-modifying antirheumatic drug (DMARD) such as hydroxychloroquine, gold, sulfasalazine, methotrexate, leflunomide or cyclosporin. Each of these DMARDs has its own distinct toxicities but has also been shown to be effective in reducing signs and symptoms of disease and to some extent, reduce radiological progression. Within the past 10 years, the combination of several traditional DMARDs has been shown to have increased efficacy over monotherapy without a significant increase in toxicity in a majority of studies. Recently, the US Food and Drug Administration has approved infliximab, a chimeric monoclonal antibody to tumour necrosis factor (TNF)-alpha in combination with methotrexate, for the treatment of signs and symptoms of RA, delay of radiological progression of disease and improvement of physical function while anakinra, an interleukin-1 receptor antagonist, has been approved for the treatment of the signs and symptoms of RA either as monotherapy or in combination with methotrexate. Etanercept is the first biological response modifier approved for use in RA in the US. Double-blind, randomised controlled studies have shown etanercept to be effective therapy in patients with RA who have had inadequate response to DMARDs, in combination with methotrexate, and as early monotherapy. Similar results were seen in juvenile and psoriatic arthritis in DMARD nonresponders. Open-label studies have shown efficacy in adult Still's disease, ankylosing spondylitis, progressive systemic sclerosis, Wegener's granulomatosis and chronic uveitis. Safety issues are a concern because of the ubiquitous role of TNF. To date the only consistent adverse event seen with etanercept has been injection site reactions. Infections occur at the same rate and with the same frequency as the placebo population. There should be caution, however, with using etanercept in patients with a serious infection, or recurrent infections or patients with untreated or latent tuberculosis. As of yet there has not been seen an increase of malignancies. Rare neurological and haematological events have been noted. Etanercept has been a significant addition to the armamentarium of medications for the treatment of RA, juvenile and psoriatic arthritis. Preliminary data show that it may be well tolerated and effective in other rheumatic diseases in which there is over production of TNFalpha. PMID- 11945117 TI - Applications of peptide mimetics in cancer. AB - The development of a vaccine for cancer has been difficult compared to the effective vaccines of infectious diseases. Most tumor antigens are not entirely foreign and are expressed on normal tissues, thus, making it difficult to induce strong immune responses against self antigens. A peptide mimic, however, may have the potential to generate greater immune responses than those induced to self peptides. In this review we discuss applications of peptide mimics for cancer immunotherapy which may ultimately prove useful in humans. PMID- 11945118 TI - Sugar amino acids and their uses in designing bioactive molecules. AB - In search of new molecular entities for discovering new drugs and materials, organic chemists are looking for innovative approaches that try to imitate nature in assembling quickly large number of distinct and diverse molecular structures from 'nature-like' and yet unnatural designer building blocks using combinatorial approach. The main objective in developing such libraries is to mimic the diversities displayed in structures and properties of natural products. The unnatural building blocks used in these assemblies are carefully designed to manifest the structural diversities of the monomeric units used by nature like amino acids, carbohydrates and nucleosides to build its arsenal. Compounds made of such unnatural building blocks are also expected to be more stable toward proteolytic cleavage in physiological systems than their natural counterparts. Sugar amino acids constitute an important class of such polyfunctional scaffolds where the carboxyl, amino and hydroxyl termini provide an excellent opportunity to organic chemists to create structural diversities akin to nature's molecular arsenal. Recent advances in the area of combinatorial chemistry give an unprecedented technological support for rapid compilations of sugar amino acid based libraries exploiting the diversities of carbohydrate molecules and well developed solid-phase peptide synthesis methods. This review describes the development of sugar amino acids as a novel class of peptidomimetic building blocks and their applications in creating large number of structurally diverse peptide-based molecules many of which display interesting three-dimensional structures as well as useful biological properties. PMID- 11945119 TI - In vitro and in vivo activities of ciprofloxacin and levofloxacin against an SHV 5 extended-spectrum beta-lactamase-producing Klebsiella pneumoniae strain. AB - Activities of ciprofloxacin and levofloxacin against an SHV-5 extended-spectrum beta-lactamase (ESBL)-producing Klebsiella pneumoniae strain were studied in vitro and in vivo in septic mice using a high inoculum. Susceptibility to ciprofloxacin and levofloxacin was independent of the inoculum size. In killing curve studies, after 24 hours the initial 7.69 log10 CFU/ml increased in the control to 9.34, while it was reduced to 4.83 by ciprofloxacin and to 4.25 by levofloxacin. Mice were infected with 10(7) CFU/g of K. pneumoniae intraperitoneally. Treatment started 2 hours later, when the mean blood bacterial counts were 7.33 log10 CFU/ml, and lasted for 26 hours from the time of infection. Blood bacterial count was reduced from 7.33 log10 CFU/ml to 4.08 log10 CFU/ml by ciprofloxacin (20 mg/kg/6 hours), and to 3.60 log10 CFU/ml by levofloxacin (50 mg/kg/6 hours) 8 hours after the infection, which differed significantly from the infected untreated group. Ciprofloxacin and levofloxacin prolonged significantly the survival of mice compared with the infected untreated group (p<0.001 for both groups). There were not significant differences either in the survival (p=1.0) or in the blood bacterial counts (p=0.216 after 8 hours) between ciprofloxacin and levofloxacin group. Based on these results both ciprofloxacin and levofloxacin could be alternative therapeutic agents for the infection caused by ESBL-producing Klebsiella strains. PMID- 11945120 TI - Arylpiperazine derivatives acting at 5-HT(1A) receptors. AB - Serotonin (5-hydroxytryptamine, 5-HT) is one of the most attractive targets for medicinal chemists. Among 5-HTRs, the 5-HT(1A) subtype is the best studied and it is generally accepted that it is involved in psychiatric disorders such as anxiety and depression. Several structurally different compounds are known to bind 5-HT(1A)R sites. Among these, arylpiperazine derivatives represent one of the most important classes of 5-HT(1A)R ligands. This article will review the development of arylpiperazine derivatives acting at 5-HT(1A)Rs with an emphasis on structure-affinity relationships of agonists and antagonists, ligand-receptor interactions and pharmacological applications. PMID- 11945121 TI - Recent developments in depsipeptide research. AB - This review focuses on the major developments in depsipeptide research since 1995. Depsipeptides are bio-oligomers composed of hydroxy and amino acids linked by amide and ester bonds. Many depsipeptides show very promising biological activities, including anticancer, antibacterial, antiviral, antifungal, anti inflammatory, and anti-clotting or anti-antherogenic properties. In this report depsipeptides exhibiting these properties are discussed. Their isolation, structural determination, and notable structural features are discussed, but their biological properties and therapeutic potentials are emphasized. Depsipeptides have shown the greatest therapeutic potential as anticancer agents. Four depsipeptides have entered clinical trials for cancer treatment. Among the antiviral compounds discovered, the callipeltins and the quinoxapeptins are particularly promising due to their inhibitory activities against HIV. These compounds have the potential to be developed as anti-AIDS drugs or to serve as lead compounds for the discovery of structurally related anti-AIDS compounds. Antifungal compounds, such as the jaspamides, may lead to therapies against many of the opportunistic infections that accompany AIDS. Anti-inflammatory compounds such as SCH217048 act as neurokinin antagonists and may lead to anti-inflammatory treatments. Some depsipeptides such as micropeptins and A90720A have been found to be effective plasmin inhibitors, which have implications as treatments for cardiovascular diseases. Compounds such as SCH58149 help control the levels of HDL and LDL. PMID- 11945122 TI - Structural basis of the dysfunctioning of human 2-oxo acid dehydrogenase complexes. AB - 2-oxo acid dehydrogenase complexes are a ubiquitous family of multienzyme systems that catalyse the oxidative decarboxylation of various 2-oxo acid substrates. They play a key role in the primary energy metabolism: in glycolysis (pyruvate dehydrogenase complex), the citric acid cycle (2-oxoglutarate dehydrogenase complex) and in amino acid catabolism (branched-chain 2-oxo acid dehydrogenase complex). Malfunctioning of any of these complexes leads to a broad variety of clinical manifestations. Deficiency of the pyruvate dehydrogenase complex predominantly leads to lactic acidosis combined with impairment of neurological function and/or delayed growth and development. Maple urine disease is an inborn metabolic error caused by dysfunction of the branched-chain 2-oxo acid dehydrogenase complex. An association between both Alzheimer disease and Parkinson s disease and the 2-oxoglutarate dehydrogenase gene has been reported. Currently a wealth of both genetic and structural information is available. Three dimensional structures of three components of the complex are presently available: of the pyruvate dehydrogenase component (E1), of the dihydrolipoyl acyltransferase component (E2) and of the lipoamide dehydrogenase component (E3). Moreover, detailed information on the reaction mechanism, regulation and the interactions between the different components of the complex is now at hand. Although only one of the structures is of human origin (E1b), model building by homology modelling allows us to investigate the causes of dysfunction. In this review we have combined this knowledge to gain more insight into the structural basis of the dysfunctioning of the 2-oxo acid dehydrogenase complexes. PMID- 11945124 TI - Endothelin receptor antagonists--an overview. AB - In thirteen years since the appearance of Endothelin (ET) on the international scene, possibility of its involvement in a variety of diseases has attracted the attention of medicinal chemists in search of novel therapeutics for various cardiovascular diseases (CVDs). Discovery of pharmaceutical agents which either block the generation of ET from its precursor or antagonize its binding to cellular receptor, should not only provide means to assess the physiological role of ET, but lead to useful therapy for conditions associated with altered production or responsiveness to ET. In this review article, we have attempted to present in a classified format, the kaleidoscope of ET receptor antagonists that have emerged through structure activity relationship studies using the parent peptide as well as from screening of various compound libraries. By all indications, the variety and range of small molecules that are currently under investigation continues to open up newer opportunities and lures fresh groups of scientists into this research arena. Presently a number of these compounds are in the clinics, being evaluated for their beneficial effects in a range of human pathologies such as essential hypertension and chronic heart failure. PMID- 11945123 TI - Schizophrenia: genesis, receptorology and current therapeutics. AB - Schizophrenia is a debilitating mental disease affecting approximately 1% of the population worldwide. Since the discovery of the first modern treatment for schizophrenia, chlorpromazine, in 1952 there have been many new structures investigated, only a small fraction of which have resulted in clinically useful drugs. Of these, haloperidol may be regarded as the drug for first line treatment. Since then, clozapine has emerged as the benchmark therapeutic ameliorating positive and negative symptoms and devoid of movement disorders, with its greatest feature being improvement of treatment-resistant patients. However, a major, potential lethal side-effect of clozapine is the induction of agranulocytosis, a blood disorder with unknown mechanism that results in lowered white-blood cell counts and consequent susceptibility to infections. In the 50 years of antipsychotic drug development, several novel theories have evolved that focus on receptor sub-types (serotonin 5-HTsub>2A, dopamine D(2) and D(4)) and the degree to which they need to be selectively attenuated by the drugs. Also of significance is the location of these receptors in the brain in relation to the disease state, the myriad of side-effects associated with antipsychotics and physicochemical properties of antipsychotic molecules relative to models of the drugs and the GPCR receptors involved. The techniques for investigation have shown increasing sophistication and refinement over this period, involving cloned receptors and PET scanning for determination of receptor location, density and binding, and rate constants at receptors. Knowledge of receptor structure, although in its infancy since no membrane bound CNS-receptor has yet been crystallized, is likely to benefit substantially with advances in computer-aided modelling. Overall, these new techniques have resulted in a number of novel antipsychotics such as risperidone, sertindole, olanzapine, seroquel, zotepine and ziprasidone, whose design, synthesis and testing has benefited enormously from the accumulated knowledge base of the past 50 years. In this review, we will provide a comprehensive update of the theories of action and clinical profiles of the latest drugs listed. The following appraisal of the literature will provide the practising medicinal chemist interested in this critical area of research with sufficient insight and understanding, to embark on productive investigations into the design and development of new therapeutic agents devoid of clinically limiting side-effects. PMID- 11945125 TI - Recent advances in antiplatelet agents. AB - Platelet aggregation plays a key role in the pathogenesis of thromboembolic diseases such as myocardial infarction, stroke, unstable angina and peripheral artery disease. Until recently, aspirin was the only antiplatelet agent available to prevent or treat these events. Over the past several years, there has been a substantial expansion in the antiplatelet armamentarium as well as in the understanding of the clinical importance of antiplatelet therapy in limiting the complications of thrombosis. Aspirin was one of the first agents to be adopted and it remains as the standard therapy with the higher amount of available clinical information. Following aspirin, ADP receptor antagonists like ticlopidine and clopidogrel as well as phosphodiesterase inhibitors dipyridamole and cilostazol have been introduced. Glycoprotein (GP) IIb/IIIa receptor antagonists like eptifibatide, tirofiban and abciximab are the newer antiplatelet agents which act at the end of the common pathway of platelet aggregation. Although results of clinical studies with the first oral GPIIb/IIIa antagonists were disappointing, agents of the new generation might expand the potential application of GPIIb/IIIa targeted therapy. This review will highlight recent advances in the development of aspirin, phosphodiesterase inhibitors, ADP receptor antagonists and the platelet glycoprotein IIb/IIIa inhibitors. The emphasis of this paper has been placed on the chemical aspects of these agents. PMID- 11945126 TI - Novel synthetic retinoids and separation of the pleiotropic retinoidal activities. AB - Retinoids, all-trans-retinoic acid (1a) and its analogs, act as specific modulators of cellular differentiation and proliferation, through binding to and activating specific nuclear receptors, retinoic acid receptors (RARs) and retinoid X receptors (RXRs). Retinoids have chemotherapeutic roles in dermatology and oncology, but their usefulness is restricted by the high toxicity of retinoic acid and its hydrophobic analogs. We have developed various retinoidal benzoic acid derivatives, and named them retinobenzoic acids. Among them, aromatic amides such as Am80 (7) and Am580 (8) have superior pharmacological characteristics, including RAR subtype selectivity. Structural modification based on the ligand superfamily concept afforded several types of RAR antagonists, benzimidazole derivatives, BIPh (41) and BIBn (42), and dibenzodiazepine derivatives, LE135 (46) and LE540 (47). LE135 (46) is a unique antagonist with RARbeta-selectivity. During investigations on the structure-activity relationships of retinobenzoic acids, several retinoid synergists (RXRs agonists), such as HX600 (49), DA113 (55h) and TZ335 (57), have been found. These compounds are expected to modulate other nuclear receptors which form heterodimers with RXRs, besides retinoids. Further, we found some RXRs antagonists, HX531 (60) and HX603 (61), which inhibit the activation of both RXR homodimers and RXR RAR heterodimers. In this review, we describe our investigations on these structurally and biologically unique retinoids and retinoid-regulatory compounds. PMID- 11945127 TI - Analysis of ligand-macromolecule contacts: computational methods. AB - Due to the many technological advancements in biology and development of new fields such as biotechnology and bioinformatics, our knowledge of cellular functions has been growing rapidly; and Biology has entered the Information Age. Along with the technological advancements has come a rapid increase in identification of biomolecular targets involved in diseases. Recently, structure based drug design studies have emphasized integration of the clinical, cellular, biochemical, structural, and biophysical knowledge of the target. Due to advances in sequencing the human genome, in chemical synthesis and structure determination of biological targets using X-ray and NMR techniques, and in high-performance computing, many scientists from both experimental and theoretical fields focus on structure-based drug design. As scientists in such wide-ranging disciplines, we must understand the data from and educate one another about the strengths and weaknesses of our various disciplines. Since 1990, we have been using computers to visually evaluate ligand binding. In this review, the author will focus on computational methods that not only visualize but also quantify the nature and strength of ligand-macromolecule contacts. Such quantification can be very useful both for medicinal chemists to design ligands and for molecular biologists to design rational protein design experiments to study the effect of amino acid changes on ligand binding. PMID- 11945128 TI - Discovery and design of retinoic acid receptor and retinoid X receptor class- and subtype-selective synthetic analogs of all-trans-retinoic acid and 9-cis-retinoic acid. AB - This review presents a historical overview of the discoveries of retinoic acid receptor (RAR) and retinoid X receptor (RXR) class- and subtype-selective synthetic retinoids. These synthetic retinoids are conformationally restricted by having aromatic rings in place of the tetraene bond systems of all-trans- and 9 cis-retinoic acids. Events leading to the design and synthesis of such retinoid transcriptional agonists as RAR subtype beta,gamma-selective 6-(5,6,7,8 tetrahydro-5,5,8,8-tetramethyl-2-naphthalenyl)-2-naph-thalenecarboxylic acid (TTNN), the RARgamma-selective Z-oxime of 6-(5,6,7,8-tetrahydro-5,5,8,8 tetramethyl-2-naphthalenylcarbonyl)-2-naphthalenecarboxylic acid (SR11254), RAR selective 4-(5,6,7,8-tetrahydro-5,5,8,8-tetramethyl-2-anthracenyl) benzoic acid (TTAB), RXR-selective 4-[1-(5,6,7,8-tetrahydro-5,5,8,8-tetramethyl-2 naphthalenyl)-cyclopropyl] benzoic acid (SR11246), RXR-selective 4-[1-(5,6,7,8 tetrahydro-3,5,5,8,8-pentamethyl-2-naphthalenyl)-2-methylpropenyl]benzoic acid (SR11345), and RARgamma-selective retinoid transcriptional antagonist 2-(6 carboxy-2-naphthalenyl)-2-(5,6,7,8-tetrahydro-5,5,8,8-tetramethyl-2-naphthalenyl) 1,3-dithiolane (SR11253) are described. PMID- 11945129 TI - Cu, Fe, Mn, and Zn chelates offer a medicinal chemistry approach to overcoming radiation injury. AB - This review points out that treatment with essential metalloelement (Cu, Fe, Mn, and Zn) chelates facilitate tissue repair processes required for recovery from radiation injury including survival of lethally irradiated mice and rats. Results of studies pertaining to successful uses of bioavailable essential metalloelement chelates and combinations of them as well as aminothiols, Ca-channel blockers, acyl Melatonin homologs, substituted anilines, and curcumin radioprotectants are included in this review to suggest their use as chelates in overcoming radiation injury. Additional reports document that non-toxic doses of essential metalloelement chelates are effective in increasing survival and repairing radiation injury when administered before irradiation, in the radiation protection paradigm, and effective in increasing survival when used to treat after irradiation, in the radiorecovery paradigm. There are no other agents known to be effective in increasing survival when they are used to treat after irradiation. These approaches to radioprotection and radiorecovery offer promising approaches to facilitating recovery from radiation-induced injury experienced by patients undergoing radiation therapy for their neoplastic disease and by individuals who experience environmental, occupational, or accidental exposure to ionizing radiation. These individuals include those exposed to radiation resulting from nuclear accidents, the use of depleted uranium missiles, and astronauts undertaking space travel. Since there are no existing safe and effective treatments of radiation injury, studies of essential metalloelement chelates and combinations of them, as well as combinations of them with existing radioprotectant aminothiols, Ca-channel blockers, acyl Melatonin homologs, substituted anilines, and curcumin as radioprotectants seem worthwhile. PMID- 11945131 TI - Human carcinogenesis and bracken fern: a review of the evidence. AB - The complex taxon embraced in the Pteridium genus, popularly known as bracken fern and notorious weeds in many parts of the world, is one of the few vascular plants known to induce cancer naturally in animals. It has been known for long to be acutely toxic to livestock and sublethal chronic oral feeding of bracken fronds leads to cancerous lesions in the urinary bladder, or bovine enzootic haematuria (BEH) and ileum of cattle. Bracken poisoning has been attributed chiefly to ptaquiloside, a norsesqui-terpene which is also a potent carcinogen inducing various malignancies in laboratory animals. It is capable of alkylating uncoiled DNAbases at key proto-oncogenes of selected organs. Some human populations also eat young bracken shoots and epidemiological studies in Japan and Brazil have shown a close association between bracken consumption and cancers of the upper alimentary tract. In addition, other studies reveal that the mere presence of bracken swards represents a greater risk to die of gastric adenocarcinoma for people who live more than 20 years in such areas or are exposed in childhood. This work reviews the bracken-cancer connections established by in vitro and in vivo experiments and epidemiological studies in various parts of the world, and provides insights into the possible bridges for bracken carcinogens to reach the human diet. Also, specific points where more research is needed are highlighted. PMID- 11945130 TI - The palmitoylethanolamide family: a new class of anti-inflammatory agents? AB - The discovery of anandamide as an endogenous ligand for the cannabinoid receptors has led to a resurgence of interest in the fatty acid amides. However, N palmitoylethanolamine (PEA), a shorter and fully saturated analogue of anandamide, has been known since the fifties. This endogenous compound is a member of the N-acylethanolamines, found in most mammalian tissues. PEA is accumulated during inflammation and has been demonstrated to have a number of anti-inflammatory effects, including beneficial effects in clinically relevant animal models of inflammatory pain. It is now engaged in phase II clinical development, and two studies regarding the treatment of chronic lumbosciatalgia and multiple sclerosis are in progress. However, its precise mechanism of action remains debated. In the present review, the biochemical and pharmacological properties of PEA are discussed, in particular with respect to its analgesic and anti-inflammatory properties. PMID- 11945132 TI - Fluorinated anthracyclines: synthesis and biological activity. AB - Organic structures with fluorine atom are slightly diffuse in nature. Starting 80s researchers have discovered that the selective introduction of fluorine into biologically active molecules exercised an influence on activity. So an important endeavour in drug design have been described and numerous compounds incorporating fluorine as either a bioisosteric replacement for hydrogen or an isoelectronic replacement for the hydroxyl group have been reported. Parallely, an enormous literature on anthracyclines exists, a class of compounds used in clinical since 70s, as antitumor drugs. Unfortunately, the anthracyclines are known as well for several toxical effects that frequently condition the clinical use. In the last decade a lot of anthracycline derivatives has been described in which has been introduced a fluorine atom in different position of molecule. This review wishes to represent an updated collection of compounds with anthracycline structure where a fluorine atom has been introduced on aglycon or/and sugar moiety. Together with the chemical structures, the synthetic indications are furnished and succinct explanations of biological activity are summarised (if available). PMID- 11945133 TI - Controlling apoptosis by inhibition of caspases. AB - The intracellular cysteine proteinases grouped under the common name of caspases are important participants in the process of programmed cell death called apoptosis. Of the nearly fourteen mammalian members discovered thus far caspase 1 or (interleukin 1beta converting enzyme; ICE), and possibly other related family members also serve as activator of cytokines. In general, caspases act on a number of cellular targets including other caspase family members leading ultimately to apopto4 4is through a highly integrated and regulated biological, biochemical and genetic mechanism. The proper execution of apoptosis is crucial during developmental stages and continues to be of critical importance for the well being of the mature organism. However, in a number of degenerative diseases the pathological states are characterized by an exacerbated loss of certain types of cells, cellular death that has morphological characteristics of apoptosis. Fortunately, it has been known for sometime that induced apoptosis that proceeds through the activation of caspases can be inhibited to rescue these cells and allow them to remain viable. This realization has attracted attention towards caspases as likely targets for pharmacological intervention, believing that inhibition of their enzymatic activity in the compromised cells will prevent the unwanted high rate of cellular death. Here we survey natural and synthetic inhibitors of caspases that have been reported to date, including some commonly used peptide inhibitors that serve as "tool reagents" in research and others that have been used to map inhibitor binding interaction in the active site. PMID- 11945134 TI - Amino-alkyl-cyclohexanes as a novel class of uncompetitive NMDA receptor antagonists. AB - Because of its widespread involvement in the physiology and pathology of the CNS, the glutamatergic system has gained considerable attention as a potential target for development of new agents for a number of therapeutic indications. In this respect, the glutamate receptor subtype of the NMDA type has been most intensively studied. The present review describes the rational for developing amino-alkyl-cyclohexanes, as new uncompetitive NMDA receptor antagonists based on our positive experience with memantine which has been used clinically for many years for the treatment of neurodegenerative dementia. Many amino-alkyl cyclohexane derivatives have been evaluated in vitro and in animal models, and in turn, one structure, namely neramexane HCl (MRZ 2/579) was selected for further development. This agent shows some similarity to memantine e.g. channel blocking kinetics, voltage dependency, and affinity. Preclinical tests indicated particularly good activity in animal models of alcoholism (self-administration, withdrawal-induced audiogenic seizures etc.) and pain (chronic pain, inhibition of tolerance to the analgesic effects of morphine). It turn, this agent has recently entered phase II of clinical trials in alcoholism after a favourable profile seen in phase I studies. PMID- 11945135 TI - NR2B selective NMDA receptor antagonists. AB - NR2B antagonists have received considerable attention in recent years. In this class of excitatory amino acid receptor antagonists NR2B antagonists have shown efficacy in neuroprotection, anti-hyperalgesic and anti-Parkinson animal models. Several groups are involved in developing these compounds as therapeutic agents and evaluating newer therapeutic targets for these agents. Until recently benzylpiperidine and phenylpiperidine templates, which were based on the structures of Ifenprodil and Eliprodil, formed the basis of most SAR in this area. A few chemical leads in this class such as CP-101,606, Ro25,6981 and PD0196860 have been identified as possible development leads which have generated significant interest in this area. In addition to the efforts of Pfizer (Parke Davis), Roche and E.Merck, several other industrial and academic research groups have continued to work in the NR2B area and recently Merck and Roche have reported new chemical leads as NR2B antagonists with significantly different biaryl templates. These new advances have raised hope, for potential success of the NR2B antagonists as new therapeutic agents, for the treatment of several pathophysiological indications. PMID- 11945136 TI - The AMPA receptor binding site: focus on agonists and competitive antagonists. AB - It is generally agreed that (S)-glutamic acid (Glu) receptors are involved in the development of a number of diseases in the central nervous system (CNS), and ligands that interact with these receptors are of significant interest. Selective ligands are indispensable as tools for the elucidation of the physiological role of AMPA receptors and as leads for the development of therapeutic agents. Over the last decade a wide variety of such ligands have been developed and studies on the structure-activity relationships of these compounds have contributed to our understanding of the mechanisms involved in AMPA receptor activation and blockade. Series of selective agonists using the 3-isoxazolol amino acid ibotenic acid (2) as a lead compound have been designed and developed. Other heterocycles, such as the uracil moiety of willardiine (6), have also proved to be highly effective bioisosteres for the distal carboxyl group of Glu. For a number of reasons, the development of competitive antagonists with therapeutic potential has been hampered for example due to the limited solubility of key heterocyclic compounds structurally unrelated to Glu. However, some problems have been overcome, and series of water-soluble, potent and selective quinoxalinediones, indenoimidazones and isatine oximes have now been developed. At the turn of the millennium the crystal structure of GluR2 co-crystallized with different AMPA receptor ligands became available, opening a new era in the design of AMPA receptor ligands on a rational basis. PMID- 11945137 TI - Kainate receptor agonists, antagonists and allosteric modulators. AB - Interest in kainate receptors has increased over the past few years. Our understanding of their physiology and pharmacology has improved markedly since their original cloning and expression in the early 1990s. For example, agonist profiles at recombinant kainate receptors have been used to identify and distinguish kainate receptors in neurons. Furthermore, the development of selective antagonists for kainate receptor subtypes has increased our understanding of the functional roles of kainate receptors in neurons and synaptic transmission. In this review we described the activity of agonists and antagonists at kainate receptors and their selectivity profiles at NMDA and non NMDA receptors. PMID- 11945138 TI - The role of ionotropic glutamate receptors in nociception with special regard to the AMPA binding sites. AB - The recent literature on the antinociceptive action of ionotropic glutamate receptor antagonists is reviewed with special emphasis on their clinical potential. Actually the glutamatergic pathways descending from the brain stem into the spinal cord may generate analgesia. However, physiologically more important is that glutamate and aspartate are apparently the main neurotransmitters along the ascending nociceptive pathways in the spinal cord. Glutamate, aspartate and their receptors can be detected in particularly high concentrations in the dorsal root ganglia and the superficial laminae (I, II) of the spinal cord. In low doses glutamate receptor antagonists only slightly elevate the threshold of the physiological pain sensation. However, they suppress the process of pathological sensitisation i.e. lowering of the pain threshold seen upon excessive or lasting stimulation of C-fibre afferents, a process that takes place during inflammation or other kinds of tissue injury. At electrophysiological level antagonists of both the NMDA- and AMPA/kainate receptors inhibit wind up i.e. lasting activation of the polymodal, second order sensory neurones in the deeper layers of the dorsal horn. During sensitisation the resting Mg(++) blockade of transmembrane Ca(++) channels is abolished, certain second messenger pathways are activated, the transcription of many genes is enhanced leading to overproduction of glutamate and other excitatory neurotransmitters and expression of Na(+) channels in the primary sensory neurones activated at lower level of depolarisation. This cascade of events leads to increased excitability of the pain pathways. NMDA antagonists are apparently more potent in experimental models of neuropathic pain, whereas AMPA antagonists are more effective in abolition of hyperalgesia seen during experimental inflammation. Clinically, of the previously known NMDA antagonists amantadine, dextromethorphan and ketamine have been tested, the latter extensively. Ketamine has been found quite active in certain cases of neuropathic pain and it reduced the opiate demand when used for postoperative analgesia. However, in other types of clinical pain their efficacy is less convincing. Not being registered there are no clinical data on the AMPA antagonists. There are, however, some investigational new drugs and some novel compounds in the stage of preclinical development which antagonise the AMPA receptors in competitive fashion or allosterically. Of the latter molecules 2,3-benzodiazepines are particularly promising. PMID- 11945139 TI - Non-competitive AMPA antagonists of 2,3-benzodiazepine type. AB - The discovery of the selective AMPA antagonist character of 2,3-benzodiazepine derivative GYKI 52466 (5) in the late eighties and the recognition of the non competitive nature of its mode of action some years later set off the world-wide search for novel class of drugs. Notably the quest to develop new antiepileptic and neuroprotective medicines, which allosterically inhibit the AMPA sensitive glutamate operated channels. This review summarises our present knowledge about the allosteric site, dubbed "GYKI site" where the 2,3-benzodiazepines are supposed to bind to. The structure-activity relationships among AMPA antagonist 2,3-benzodiazepines and their structural analogues with similar biological profile are reviewed in a possibly comprehensive fashion. The chemical synthesis of 2,3-benzodiazepines is shortly described. The in vitro and in vivo experimental methods used for pharmacological characterisation of the biologically active compounds are briefly explained. Finally the therapeutic potential of 2,3-benzodiazepines i.e. the main fields of their clinical utility are outlined with special regard to talampanel (20) in the light of the ongoing clinical trials with this new drug candidate. PMID- 11945140 TI - AMPA receptor antagonists with additional mechanisms of action: new opportunities for neuroprotective drugs? AB - Ischaemic stroke of the brain accounts for about one third of all deaths in industrialized countries. Many of the patients who survive are severily impaired. Thus, there is an enormous need for pharmacotherapy for the treatment of ischaemic stroke. Why is such a treatment not available yet? Are the pathophysiological sequelae of brain ischaemia not well understood? Have there been no attempts for clinical development of neuroprotective drugs? Everyone who is engaged in stroke research knows that the opposite is true: The cellular processes occuring after brain ischaemia have been studied for a long time, and we have a thorough understanding of the cellular processes which are involved. Many compounds underwent clinical trials, but most of them failed. One hypothesis to explain this disappointing fact might be that the cellular consequences of stroke are manyfold, but that the clinically tested compounds were selective for only one molecular mechanism. The aim of this review is to give a summary of the pathophysiological mechanisms which occur during and after an ischaemic stroke, and to comment on the preclinical studies where multiple disease-related mechanisms were targeted pharmacologically. Moreover, a novel class of neuroprotective compounds, the oxadiazole derivatives, will be presented. Compounds of this chemical class target two key mechanisms which are important for the pathophysiology of stroke, namely voltage-gated sodium channels, as well as glutamate receptors of the alpha-amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid (AMPA) subtype. PMID- 11945141 TI - Treatment of chronic hepatitis B. AB - Infection with the hepatitis B virus has switched over the last 20 years from the classical HBeAg positive serologic pattern to a HBeAg negative form that is linked, in the Mediterranean basin, with the epidemiologic replacement of the causative wild-type of virus B with mutant variants, whereby mutations in the core-promoter and in the pre-core region prevent the secretion of HBeAg. The wild type pattern of infection (characterized by relatively high steady level ALT, high HBV-DNA levels and clinically overt liver disease) responds relatively well to Interferon: 3 to 5 mega units daily or 10 mega units every other day for 16 weeks induce anti-HBe seroconversion, normalize the ALT and possibly also eliminate the HBsAg in some 40% of the adults with a minimal (7%) risk of relapse. However, the mutant type infection (anti-HBe positive / HBe Ag negative) is less responsive to Interferon; this has led to the search for novel nucleoside analogues which has currently culminated in the advent of Lamivudine. This competitor of cytidine is 80% bioavailable and devoid of side-effects at the oral dose of 100 mg daily; tolerance continues for therapies up to 3 years. Lamivudine therapy shares with Interferon a rapid decline of ALT accompanied by improvement of histology; at variance with Interferon there is a delayed accumulating seronconversion to anti-HBe and the switch to anti-HBs is rare. Over the long term its activity is abolished by the emergence of specific viral mutations (YMDD mutants) that rekindle the disease. The indications to Lamivudine therapy in HBeAg negative chronic hepatitis B are currently under investigation. Lamivudine is highly efficacious in preventing HBV reinfection in liver transplants. PMID- 11945142 TI - New therapies for the treatment of chronic hepatitis C. AB - Treatment of chronic hepatitis C (HCV) was based on Interferon alpha IFNalpha administration three times a week (tiw), but the efficacy of this schedule (evaluated as virological sustained response) was limited to less than 20% of patients. The combination of Ribavirin and IFN is known to be significantly more effective than IFN monotherapy in naive and relapser patients but it induces a sustained response only in 41% of patients and in less than 30% of patients infected with genotype 1. Several studies on IFN and viral kinetics suggested that daily administration of IFN may increase the sustained response rate. The development of pegylated IFNs, characterized by a long half life and weekly administrability, seems to induce a significant improvement in the treatment of chronic hepatitis particularly in combination with Ribavirin. In order to further improve the efficacy of treatment in chronic hepatitis C (HCV) many controlled clinical trials evaluating Amantadine, Micophenolate, alpha1 Thymosin, Maxamine (in combination with IFN or pegylated IFN), Protease, Helicase, Polymerase inhibitors, Ribozymes and anti-sense olygonucleotides, and Interleukin 10 are in progress. Finally anti-HCV vaccine development seems to be very promising. PMID- 11945143 TI - The treatment of chronic hepatitis C not responding to interferon. AB - Approximately 60% of all patients with chronic hepatitis C (C-HCV) treated with standard interferon (IFN) treatment, i.e. combination of recombinant alpha IFN and ribavirin (RBV), are refractory to treatment. Many factors should be responsible for HCV persistence after antiviral treatment. Beside the well-known importance of some factors such as viral heterogeneity, co-infections with Hepatitis B Virus (HBV) or Human Immunodeficiency Virus (HIV), presence or absence of fibrosis, age, sex, iron overload, a greater attention is being paid to the study of viral kinetics. Observing the trend of the slope of viral decline, already after a few hours antiviral administration, it is possible to predict the sustained virologic response and therefore to optimize therapy. As for alternative therapeutics, re-treatment with IFN alone was excluded considering the very disappointing results, whereas it seemed that the combination IFN plus RBV could recover up to 30% of the patients. Later both randomized trials and two metanalyses have demonstrated that this option is disadvantageous from the cost-effectiveness point of view since 14 patients need to be treated to obtain one responsive. The treatment combining IFN plus RBV and amantadine seems more promising. Recently trials with pegylated IFN have started with the aim to increase the therapeutical response in this category of HCV positive patients. PMID- 11945144 TI - Incidence of side effects during therapy with different types of alpha interferon: a randomised controlled trial comparing recombinant alpha 2b versus leukocyte interferon in the therapy of naive patients with chronic hepatitis C. AB - BACKGROUND: Alpha interferon (IFN) alone or in combination with Ribavirin (RBV) is the treatment of choice for HCV related chronic liver disease. There are many types of alpha IFN and to date only few reports are available comparing different types of alpha interferon. We run a randomised controlled trial with the aim to compare tolerability and efficacy of two different types of IFN: recombinant alpha 2b interferon (IFN-R) and leukocyte alpha n-3 interferon (IFN-L) at the same dosage of 3 MU subcutaneously thrice weekly for one year. METHODS: one hundred sixty eight consecutive anti-HCV positive naive patients, 34 mild chronic active hepatitis (MCH), 81 moderate-severe hepatitis (MSCR) and 53 active cirrhosis (CIRR) that met the inclusion criteria were enrolled into the study. The diagnosis of HCV chronic liver disease was established by liver biopsy performed on patients with abnormal serum alanine aminotransferase (ALT) value for at least one year. HCV serology: all patients were tested for confirmatory test RIBA II, HCV-RNA, and identification of viral genotype. Patients were randomised to receive either IFN-R or IFN-L. Follow-up continued for at least two years after stopping treatment. RESULTS: no significant differences were observed between the two groups of treatment as far as the incidence of side effects is concerned. Tolerability was good: only 11 in IFN-R and 8 patients IFN-L group respectively had to stop therapy due to side effects. The two types of IFN showed a comparable efficacy: an end of therapy response was observed in 34% of IFN-R and 30% of IFN-L patients; a sustained response was seen in 16% of IFN-R and in 19% of IFN-L patients. CONCLUSIONS: in the treatment of patients with chronic hepatitis C there was no statistically significant difference in tolerability and efficacy between the two IFNs tested. PMID- 11945145 TI - Modifications of the immune responsiveness in patients with hepatitis C virus infection following treatment with IFN-alpha/ribavirin. AB - The balance between T helper (h)1 and Th2 responsiveness seems to represent a key event in the evolution of hepatitis C virus (HCV) infection. In particular, Th1 cytokines [interleukin (IL-2) and interferon (IFN-gamma)] have been demonstrated to mediate the antiviral immune response. Serum levels of Th1 cytokines (IL-2 and IFN-gamma) as well as of Th2 products (IL-4 and IL-10) were determined in a group of HCV-positive patients before and after treatment with IFN-alpha and Ribavirin (RIB). Results indicate that responder patients exhibited increased levels of IFN gamma and IL-10, while this enhancement was not observed in non-responder patients. In this respect, the major effect exerted by the combined therapy with IFN-alpha/RIB could be represented by the attainment of a re-equilibrium between inflammatory (Th1) and antiinflammatory (Th2) mechanisms. In this framework, according to current literature, novel therapeutical approaches to treat HCV infection are represented by administration of recombinant IL-2 and IL-10. PMID- 11945146 TI - Biological and clinical significance of endotoxemia in the course of hepatitis C virus infection. AB - Endotoxins or lipopolysaccharides (LPS), major components of the cell wall of Gram-negative bacteria, once released from the bacterial outer membrane bind to specific receptors and, in particular, to a membrane-bound receptor, the CD14 (mCD14) and the toll-like receptor 4 present on monocytes/ macrophages. In turn, LPS-activated monocytes/ macrophages release in the host tissue an array of so called proinflammatory cytokines and, among them, Tumor Necrosis Factor (TNF) alpha, interleukin (IL)-1beta, IL-6, IL-8 and IL-12 are the major mediators. Before therapy (To) and at the end of 6-month interferon (IFN)-alpha/Ribavirin (RIB) treatment (T6), circulating endotoxin levels were measured in responder and non responder HCV+ patients. At T0, 57% of the non responders were endotoxin positive and had, on average, 54 pg/ml of plasma LPS while in 50% of the responder patients endotoxin were found with an average of 29 pg/ml. At T6, in responders LPS were no longer detectable, while in 42% of the non responders LPS were found (average levels 45 pg/ml). In terms of serum cytokine concentration, at T6 IFN-gamma levels when compared to those detected at T0 were increased in both endotoxin-positive and endotoxin-negative patients. However, at T6 IL-10 concentration was significantly increased only in the group of endotoxin-negative subjects (responder patients), in comparison to T0 values. The origin of endotoxemia in HCV+ patients seems to be multifactorial, likely depending on impaired phagocytic functions and reduced T-cell mediated antibacterial activity. In these patients, however, one cannot exclude the passage of LPS from the gut flora to the blood stream, owing a condition of altered intestinal permeability. At the same time, a less efficient detoxification of enteric bacterial antigens at the hepatic level should be taken into consideration. Finally, novel therapeutic attempts aimed to neutralize LPS in the host are discussed. PMID- 11945147 TI - Main characteristics of hepatocellular carcinoma and cirrhosis and therapeutic approaches. AB - Between 1995 and 1997 we studied 100 patients with hepatocarcinoma (HCC) and cirrhosis. Of these 74 were males and 26 females with a mean age of 66 years. 13% patients were only HbsAg positive, 75% only anti-HCV positive, 6% HbsAg and anti HCV and the etiology in 6% of cases was alcoholic. Alpha-foetoprotein was >400 ng/ml in only 18% of cases and portal thrombosis was present in 12%. Mononodular HCC was observed in 63% of cases (small HCC in only 38%) and in 79% was localized to the right lobe. Of the mononodular types, 70% were shown by echography to be hypoechoic, 6% hysoechoic, 6% hyperechoic and 17% mixed patterns. Histologically, 49% were well-differentiated, 45% moderately-differentiated and 6% poorly differentiated. No correlation was found between histologic pattern and number of nodules. Well-differentiated HCC was found in 51% of mononodular types and in 46% of multinodular types. Moderately-differentiated HCC was detected in 46% and 43% respectively and poorly-differentiated HCC in 3% and 11% respectively. No correlation was found between number of nodules and the degree of Edmonson. PMID- 11945148 TI - Tamoxifen in the treatment of hepatocellular carcinoma: 5-year results of the CLIP-1 multicentre randomised controlled trial. AB - BACKGROUND: In 1998, when data of a meta-analysis on tamoxifen in the treatment of hepatocellular carcinoma (HCC) had suggested a little advantage for this treatment, we published the results of a multicenter randomised controlled trial, that showed no survival benefit for tamoxifen vs. control. Here we report an updated analysis of the study results 4.5 years after the closure of enrollment. METHODS: The study had a planned sample size of 480 patients. Patients with any stage HCC were eligible, irrespective of locoregional treatment. Tamoxifen was given orally, 40 mg/die, from randomisation until death. RESULTS: 496 patients were randomised by 30 Institutions from January 1995 to January 1997. Information was available for 477 patients. As of July 2001, 374 deaths (78%) were recorded, and median survival times were 16 and 15 months (p=0.54), in the control and tamoxifen arm. Data were further analysed separately for advanced patients and for those eligible to potentially curative locoregional treatments: relative hazard of death for patients receiving tamoxifen was equal to 0.98 (95% CI 0.76 1.25) for the former group and 1.38 (95% CI 0.95-2.01) for the latter. The prognostic score recently devised by our group (CLIP score) was, as expected, strictly correlated (p<0.0001) to the locoregional treatment received and strongly correlated with prognosis. CONCLUSIONS: the update of the present study confirms that tamoxifen is not effective in prolonging survivals, both in advanced patients and in those potentially curable and that the CLIP score is able to predict prognosis. PMID- 11945149 TI - COX selectivity and animal models for colon cancer. AB - Early experiments performed during 1980s and 1990s using carcinogen-induced rat intestinal tumor models demonstrated the inhibitory effects of non-steroidal anti inflammatory drugs (NSAIDs) on intestinal tumorigenesis. Furthermore, epidemiological studies and clinical trials for familial adenomatous polyposis (FAP) patients supported the possibility that NSAIDs can be used as chemopreventive agents. The major target molecules of NSAIDs are cyclooxygenases (COX), which catalyze the rate-limiting step of prostaglandin biosynthesis. Two isoenzymes of COX have been identified; COX-1 and COX-2. Whereas COX-1 is expressed constitutively in most tissues and responsible for tissue homeostasis, COX-2 is inducible and plays an important role in inflammation and intestinal tumorigenesis. A genetic study using compound mutant mice of COX-2(-)/(-), and Apc(Delta716) which is a model for human familial adenomatous polyposis (FAP), directly demonstrated that induction of COX-2 is critical for intestinal polyp formation. Numerous studies have also demonstrated that COX-2 selective inhibitors suppress intestinal polyp formation in Apc gene-mutant mice, and xenografted cancer cell growths. In addition, stimulation of angiogenesis is one of the major effects by COX-2 expression that is induced in the polyp stromal cells. On the other hand, another study indicated that COX-1 also plays an important role in the early stage of intestinal tumorigenesis. These data from animal model studies should be helpful in understanding the in vivo mechanism(s) of tumor suppression by NSAIDs or COX-2 inhibitors. Here, we review the animal studies that have been published as of August 2001, and reported to suppress intestinal tumor growths by NSAIDs or COX-2 inhibitors. PMID- 11945150 TI - The role of cyclooxygenase inhibitors in cancer prevention. AB - Carcinogenesis results from the long-term accumulation of genetic and epigenetic aberrations at the molecular level, which are under constant selection pressure for growth advantage. Recognizing that cancer is the result of this long-term, multi-step process provides opportunities for molecularly targeted cancer prevention. Ideally, chemopreventive agents should be low in toxicity, morbidity, and cost. Several individual agents and agent combinations are currently under evaluation in the U.S. National Cancer Institute s (NCI) chemoprevention agent development program. Nonsteroidal anti-inflammatory drugs (NSAIDs) that inhibit cyclooxygenase (COX) -1 and -2 are among the most promising classes of agents for targeted molecular prevention. PMID- 11945151 TI - Comparative analgesia, cardiovascular and renal effects of celecoxib, rofecoxib and acetaminophen (paracetamol). AB - Comparisons are made between the specific COX-2 inhibitors, celecoxib and rofecoxib, and acetaminophen. The specific COX-2 inhibitors are a significant advance in therapy because their anti-inflammatory, analgesic and antipyretic activities are associated with a high degree of gastrointestinal safety. Acetaminophen is often not considered to be a potent inhibitor of COX-2 but it is a potent inhibitor of prostaglandin synthesis in intact cells after stimulation by cytokines. Its weak activity on the pathway of prostanoid synthesis involving COX-1 is shown by its weak anti-platelet activity and good gastrointestinal safety. The specific COX-2 inhibitors and acetaminophen are analgesic after dental surgery, orthopedic surgery and in osteoarthritis although acetaminophen appears to be a slightly weaker agent. The apparent analgesic activity of both the COX-2 inhibitors and acetaminophen may, in part, be due to their anti inflammatory properties. Both groups of drugs also decrease the urinary excretion of prostacyclin metabolites consistent with inhibition of the systemic and renal activity of the COX-2 system. During repeated dosage with the specific COX-2 inhibitors, the 24 hour urinary excretion of sodium is only inhibited for the first day of treatment while the excretion of sodium is still decreased over the first 3 hours after the individual doses. Therapeutic doses of the COX-2 inhibitors and overdoses of acetaminophen have been associated with the development of occasional cases of acute renal failure. Acetaminophen also may decrease the excretion of sodium and the reason for its greater renal safety at therapeutic doses is unclear. Myocardial infarction has also been attributed to the specific COX-2 inhibitors from meta-analysis of large scale clinical trials and examination of reports of adverse drug reactions although this is still a topic of considerable discussion. No such associations have been made with acetaminophen, possibly because it is a weak inhibitor of COX-1 in platelets. PMID- 11945152 TI - Gastrointestinal safety of selective COX-2 inhibitors. AB - It appears that selective Cox-2 inhibitors do not affect the gastro duodenal mucosa whilst having anti-inflammatory and analgesic efficacy similar to non selective NSAIDs. Two broad categories of drugs are Cox-2 selective: coxibs and a number of pre-existing NSAIDs retrospectively found to have selectivity. Cox-2 inhibitors cause less dyspepsia than NSAIDs. They spare gastrointestinal mucosal generation of prostaglandins (PGs) and PG-dependent bicarbonate secretion. Coxibs cause no acute mucosal injury in endoscopic ulcers compared to NSAID comparators. In the VIGOR study all upper GI events were reduced from 4.5 per 100 patient years to 2.1 per 100 patient years with supra-therapeutic doses of rofecoxib compared with naproxen. In the CLASS study, over a period of 3 days to 6 months, incidence of ulcer complications was 0.76% with celecoxib and 1.45% for ibuprofen or diclofenac. The less substantial reduction in events in the CLASS study compared with the VIGOR may be due, at least in part, to the fact that 21% of the patients were also on low dose aspirin. However it is premature to say that the benefit of Cox-2 inhibitors is lost in patients taking aspirin. There is continuing debate on the role of Cox-2 inhibitors in patients who have other risk factors for complicated ulcer disease e.g. patients who are elderly, on aspirin or corticosteroids, have a previous ulcer or have H. pylori infection. PMID- 11945153 TI - Trypsin and chymotrypsin inhibitors in insects and gut leeches. AB - The constant increase of life expectancy is associated with major ageing of developed populations. This indicates that the new century will have one of most epidemic progressions of cardiovascular, cancer and inflammatory diseases. The high challenge for medical research is to compress such morbidity. In these conditions, invertebrates have demonstrated to be truly useful models in drug discovery for such ageing diseases. The last decade, drug discovery in leeches has opened the gate of new molecules to treat emphysema, coagulation, inflammation, dermitis and cancer. Also other invertebrates such as insects, which evolved from the annelids, harvest potential interesting molecules, such as serine protease inhibitors that can be exploited by the medical industry. PMID- 11945154 TI - Leech thrombin inhibitors. AB - Serine proteases (SP), such as thrombin, factor Xa, elastase, trypsin are implicated in many clinical disorders such as emphysema, arthritis and cardiovascular diseases. These enzymes, in normal physiological conditions, are regulated by naturally occurring serine protease inhibitors, such as anti thrombin III involved in thromb in inhibition. Primitive parasitic invertebrates have co-evolved highly specific mechanisms to communicate with their hosts for survival purposes, by blocking host processes such as blood coagulation. Thus a battery of new powerful molecules from blood-sucker animals acting at different points of the coagulation cascade such like factor Xa, thrombin, platelets aggregation inhibitors have been isolated and are now at a clinical level. In this review, we focus our attention on thrombin inhitors. PMID- 11945155 TI - The role of protease inhibition with emphasis on the effects of inflammation and vascular immune phenomena. AB - This review discusses the role of protease inhibition with emphasis on the effects of inflammation and vascular immune phenomena in non-immunocompromised hosts. A vast body of knowledge elucidating the role of protease inhibition and hemostasis has accumulated in recent years. The two subjects are intimately linked, but the focus of this review is limited to the anti-inflammatory effects of protease inhibitors. In light of the particular expertise of the present authors, this review will focus on human studies and often cite work related to open-heart surgery, since in recent years this is the area in which a large effort has been concentrated worldwide. PMID- 11945156 TI - The peptidase zymogen proregions: nature's way of preventing undesired activation and proteolysis. AB - Since the initial description of a peptidase activity, namely pepsin, in the middle of the 19(th) century, our understanding of the molecular basis of peptidase function and activation has greatly improved. Further, by sequencing entire genomes, we have reached a stage whereby it is now possible to appreciate the tremendous diversity and unique specificities of peptidases. Because of their importance in most if not all vital processes of the cell including ultimately its death, their activities must be carefully localized and kept under tight control. In addition to endogenous inhibitors, control of enzymatic activity can be achieved through their synthesis and transport as inactive zymogens. This review article will focus on the characteristics as well as the role of the proregion contained within the peptidase zymogen structure. It will survey novel zymogen structures determined in the past 5 years as well as those of selected emerging peptidase families for which there exists as yet no or little structural data. These include members belonging to the caspase, ADAMS, TTSP, MMP, Aspartyl peptidases and convertase families. PMID- 11945157 TI - Fatty acid amide hydrolase, an enzyme with many bioactive substrates. Possible therapeutic implications. AB - During the last eight years a number of bioactive lipid mediators, the amides or esters of long chain fatty acids, have been discovered or re-discovered. These are: anandamide (N-arachidonoyl-ethanolamine, AEA) and 2-arachidonoylglycerol (2 AG), two endogenous agonists of cannabinoid receptors; oleamide (cis-9 octadecenoamide), a putative endogenous sleep-inducing factor; N-palmitoylethanol amine (PEA), a compound with promising anti-inflammatory and immune-modulatory activity. These compounds are all substrates for the same hydrolytic enzyme, fatty acid amide hydrolase (FAAH), whose molecular characterization was obtained in 1996. The molecular and enzymatic properties, tissue distribution, substrate recognition properties, physiological regulation and biological role of FAAH are discussed in this article, with special emphasis on the possible pharmacological manipulation of the activity of this enzyme with therapeutic purpose. PMID- 11945158 TI - Inhibitors of the subtilase-like pro-protein convertases (SPCs). AB - Following protein biosynthesis, some of the most important cellular mechanisms that generate biological diversity are the enzymatically driven post translational modifications that ultimately lead to the formation of bioactive molecules. Within the secretory pathway, a multitude of precursor proteins are thus modified resulting in hormones, neuropeptides, growth factors, receptors and even enzymes. These modifications include cleavage at specific sites through endo or exo-peptidase action, amidation, glycosylation and sulfation. In recent years, an important family of these processing enzymes was discovered and characterized. The so-called proprotein convertases are the products of seven distinct genes and function as endopeptidases that cleave protein precursors C terminal to basic residue sites. They are structurally related to the bacterial subtilisin family of enzymes and are thus referred to as the subtilisin-like proprotein convertases (SPCs). Many studies have examined the inhibition of this family of enzymes, through the search of endogenous inhibitors or through the development of peptidyl, non-peptidyl or protein inhibitors. Some potent inhibitors have been discovered or engineered. While it is certain that potent inhibitors could serve as important tools to further elucidate the specific functions of each SPC, it has also been suggested that such inhibitors may be developed into lead compounds that could have important therapeutic applications. This review examines the progress made in regards to endogenous and engineered inhibitors and evidence for possible uses as molecular tools or in therapeutic applications. It is noted that although important inhibitory potencies have often been reported, there is generally insufficient evidence to demonstrate high levels of specificity. It is thus suggested that an important short-term challenge before the field will be a better understanding of the catalytic specificity of each SPC. PMID- 11945159 TI - Peptide and non-peptide HIV fusion inhibitors. AB - Fusion of the HIV envelope with the target cell membrane is a critical step of HIV entry into the target cell. The HIV envelope glycoprotein gp41 plays an important role in the fusion of viral and target cell membranes and serves as an attractive target for development of HIV fusion inhibitors. The extracellular domain of gp41 contains three important functional regions, i.e. fusion peptide (FP), N- and C-terminal heptad repeats (NHR and CHR, respectively). The FP region is composed of hydrophobic, glycine-rich residues that are essential for the initial penetration of the target cell membrane. NHR and CHR regions consist of hydrophobic residues, which have the tendency to form alpha-helical coiled coils. During the process of fusion of HIV or HIV-infected cells with uninfected cells, FP inserts into the target cell membrane and subsequently the NHR and CHR regions change conformations and associate with each other to form a fusion-active gp41 core. Peptides derived from NHR and CHR regions, designated N- and C-peptides, respectively, have potent inhibitory activity against HIV fusion by binding to the CHR and NHR regions, respectively, to prevent the formation of the fusion active gp41 core. C-peptide may also bind to FP, thereby blocking its insertion into the target cell membrane. One of the C-peptides, T-20, which is in the phase III clinical trials, has potent in vivo activity against HIV infection and is expected to become the first peptide HIV fusion inhibitory drug in the near future. However, this peptide HIV fusion inhibitor lacks oral availability and is sensitive to the proteolytic digestion. Therefore, it is essential to develop small molecular non-peptide HIV fusion inhibitors having a mechanism of action similar to the C-peptides. One of the approaches in identifying the inhibitors is to use an immunological assay to screen chemical libraries for the compounds that potentially block the interaction between the NHR and CHR regions to form a fusion-active gp41 core. In combination with computer-aided molecular docking techniques, the first active non-peptide HIV fusion inhibitor targeting gp41, ADS J1, was identified. Other potential candidates of non-peptide HIV fusion inhibitors have also been identified using different approaches. It is expected that both peptide and non-peptide HIV fusion inhibitors will be developed as new classes of anti-HIV drugs, which will be used alone or in combination with HIV reverse transcriptase and protease inhibitors, for the treatment of HIV infection and AIDS. PMID- 11945160 TI - Trends in the design of nucleoside analogues as anti-HIV drugs. AB - HIV infection remains incurable although several anti-HIV drugs have been identified and developed. Among these the nucleoside analogues were and remain in the forefront of anti-HIV chemotherapeutic regimens. Most of these nucleoside analogues are modified mainly in the sugar moiety. In general, they lack a free hydroxy group at the 3'-position. Consequently, they cannot participate in the formation of a 3',5'-phosphodiester linkage, which renders the nucleoside 5' triphosphates of such nucleoside analogues effective anti-HIV agents. Preventing the formation of 3',5'-phosphodiester linkages leads to inhibition of the viral DNA strand elongation and ultimately chain termination. The phosphorylation of nucleoside analogues is a key factor in their efficacy as anti-HIV agents. Efficient phosphorylation depends largely on the structure of the nucleoside. Therefore, modification of the structure of nucleoside analogues are sought to enhance their phosphorylation and ultimately, effective inhibition of the HIV reverse transcriptase. This review is concerned with the trends in the design of nucleoside analogues as anti-HIV agents. PMID- 11945161 TI - Inhibitors of HIV-1 reverse transcriptase and integrase: classical and emerging therapeutical approaches. AB - The rapid spread of the AIDS epidemic has stimulated the search for new agents able to arrest the replication of the causative virus, HIV. The best strategy for AIDS treatment involves a combination therapy using inhibitors of reverse transcriptase and protease. However, the emergence of HIV-1 strains resistant to these drugs and their cytotoxicity requires the synthesis and the biochemical and cellular characterization of new antiviral drugs, as well as the development of newer strategies and viral targets. In addition to reverse transcriptase and protease, other retroviral enzymes acting in the replicative cycle of HIV-1 are potential targets for chemotherapeutic intervention. Like all retroviruses, HIV-1 requires the integration of the proviral double-stranded DNA, arising from the reverse transcription step, into the host chromosome for its efficient replication, maintenance of a stably infected state and productive infection. DNA integration is carried out by integrase so this enzyme represents a key area in developing new anti-retroviral therapy. Another novel enzymatic target concerns the RNase H activity associated with the retroviral reverse transcriptase, since a functional RNase H is essential for retroviral replication. Inhibitors against HIV-1 integrase and RNase H having potential therapeutical propeties have not yet been described. We focus this review on the properties of inhibitors of reverse transcriptase and integrase. Some of these antiviral agents have been known for several years while others are emerging as new promising strategies based on the use of oligonucleotides with special emphasis on the SELEX approach, peptides and retrovirucides. PMID- 11945162 TI - Non-nucleoside HIV-1 reverse transcriptase (RT) inhibitors: past, present, and future perspectives. AB - Along with nucleoside reverse transcriptase inhibitors (NRTIs) and protease inhibitors (PIs), non-nucleoside reverse transcriptase inhibitors (NNRTIs) have gained a definitive and important place in the treatment of HIV-1 infections, and are in rapid development. These compounds can be grouped into two classes: the first generation NNRTIs, mainly discovered by random screening, and the second generation NNRTIs, developed as a result of comprehensive strategies involving molecular modelling, rationale-based drug synthesis, biological and pharmacokinetic evaluations. The recent boom of NNRTIs is mainly due to their antiviral potency, high specificity and low toxicity. The rapid emergence of drug resistant HIV-1 strains induced by the first generation drugs is a disadvantage bypassed, in part, by the broad spectrum second generation NNRTIs. Starting from the first generation, this review will focus on the second generation NNRTIs dealing with the recent and most interesting published results, highlighting the guidelines for the development of a third generation of NNRTIs. PMID- 11945163 TI - Applications of artificial neural network in AIDS research and therapy. AB - In recent years considerable effort has been devoted to applying pattern recognition techniques to the complex task of data analysis in drug research. Artificial neural networks (ANN) methodology is a modeling method with great ability to adapt to a new situation, or control an unknown system, using data acquired in previous experiments. In this paper, a brief history of ANN and the basic concepts behind the computing, the mathematical and algorithmic formulation of each of the techniques, and their developmental background is presented. Based on the abilities of ANNs in pattern recognition and estimation of system outputs from the known inputs, the neural network can be considered as a tool for molecular data analysis and interpretation. Analysis by neural networks improves the classification accuracy, data quantification and reduces the number of analogues necessary for correct classification of biologically active compounds. Conformational analysis and quantifying the components in mixtures using NMR spectra, aqueous solubility prediction and structure-activity correlation are among the reported applications of ANN as a new modeling method. Ranging from drug design and discovery to structure and dosage form design, the potential pharmaceutical applications of the ANN methodology are significant. In the areas of clinical monitoring, utilization of molecular simulation and design of bioactive structures, ANN would make the study of the status of the health and disease possible and brings their predicted chemotherapeutic response closer to reality. PMID- 11945164 TI - Strategies for new antimicrobial proteins and peptides: lysozyme and aprotinin as model molecules. AB - The increasing development of bacterial resistance to traditional antibiotics has reached alarming levels, thus necessitating the strong need to develop new antimicrobial agents. These new antimicrobials should possess both novel modes of action as well as different cellular targets compared with the existing antibiotics. Lysozyme, muramidase, and aprotinin, a protease inhibitor, both exhibit antimicrobial activities against different microorganisms, were chosen as model proteins to develop more potent bactericidal agents with broader antimicrobial specificity. The antibacterial specificity of lysozyme is basically directed against certain Gram-positive bacteria and to a lesser extent against Gram-negative ones, thus its potential use as antimicrobial agent in food and drug systems is hampered. Several strategies were attempted to convert lysozyme to be active in killing Gram-negative bacteria which would be an important contribution for modern biotechnology and medicine. Three strategies were adopted in which membrane-binding hydrophobic domains were introduced to the catalytic function of lysozyme, to enable it to damage the bacterial membrane functions. These successful strategies were based on either equipping the enzyme with a hydrophobic carrier to enable it to penetrate and disrupt the bacterial membrane, or coupling lysozyme with a safe phenolic aldehyde having lethal activity toward bacterial membrane. In a different approach, proteolytically tailored lysozyme and aprotinin have been designed on the basis of modifying the derived peptides to confer the most favorable bactericidal potency and cellular specificity. The results obtained from these strategies show that proteins can be tailored and modelled to achieve particular functions. These approaches introduced, for the first time, a new conceptual utilization of lysozyme and aprotinin, and thus heralded a great opportunity for potential use in drug systems as new antimicrobial agent. PMID- 11945165 TI - Tailoring an antibacterial peptide of human lysosomal cathepsin G to enhance its broad-spectrum action against antibiotic-resistant bacterial pathogens. AB - Neutrophils contain several cationic antimicrobial proteins or peptides (CAPs) that exert antibiotic-like action against bacteria. These host-derived antibiotics kill susceptible bacteria by oxygen-independent mechanisms. Considerable interest in their activity has been generated in recent years due not only to their likely important role in innate host defense against infection, but also their possible use as therapeutic agents in treating infections caused by antibiotic-resistant pathogens. We have studied the antibacterial properties of human lysosomal cathepsin G (cat G). This highly cationic serine protease contains at least three antibacterial regions that by themselves can exert antibacterial action against Gram-negative bacteria, such as Pseudomonas aeruginosa. Only one of these peptides, defined by residues 117-136 of full length cat G, has bactericidal action against Gram-positive pathogens, such as Staphylococcus aureus. Due to the broad-spectrum antibacterial action of this peptide, we have sought to define the amino acids within its primary sequence required for this activity and have developed variants with improved activity. This review emphasizes the importance of both cationicity and hydrophobicity as necessary characteristics for the antibacterial action of CAPs. It also proposes the strategy that naturally occurring large human CAPs can be dissected to smaller CAPs and then modified to enhance their activity in vitro. This approach could prove beneficial to those interested in developing antimicrobial peptides as therapeutic agents. PMID- 11945166 TI - Cationic peptides: distribution and mechanisms of resistance. AB - Cationic antimicrobial peptides are observed throughout nature. In mammals they are observed both at epithelial surfaces and within the granules of phagocytic cells. They are an important component of innate defences, since in addition to their ability to kill microorganisms, they are able to modulate inflammatory responses. With respect to their ability to kill bacteria, it is very difficult to isolate resistant mutants. However there are a few known mechanisms of intrinsic resistance, including PhoPQ-dependent and other alterations in lipopolysaccharide structure that influence self promoted uptake, and protease mediated resistance. PMID- 11945167 TI - From innate immunity to de-novo designed antimicrobial peptides. AB - Antimicrobial peptides are a large group of gene-encoded, net positively charged polypeptides, produced by living organisms of all types including human and plants. They are mobilized shortly after infection as part of the innate immunity of these species and act rapidly to neutralize a broad range of microbes. Nowadays, thousands of native and de-novo designed antimicrobial peptides are available. They vary considerably in length, composition, charge and secondary structure. Despite these variations most antimicrobial peptides use a similar target, which is the bacterial phospholipid membrane. Many of them use a common general mechanism, the carpet mechanism, in which they accumulate on the bacterial membrane up to a threshold concentration, and then effect membrane permeation/disintegration. However, the structure of the permeation pathway may vary for different peptides and may include channel aggregates, toroidal pores or channels. Target specificity is determined by the negatively charged bacterial membrane, the net positive charge of the peptide, its hydrophobicity, oligomeric state in solution and in the membrane, and the stability of its secondary structure. A novel group of non hemolytic antimicrobial peptides were derived from diastereomers (containing D- and L-amino acids) of lytic peptides based on parameters required by the carpet mechanism. Because these disastereomers exhibit several advantages over their all-L amino acid counterparts, they have a potential to be developed for therapeutic use both in vitro and in vivo. PMID- 11945168 TI - Host-defense antimicrobial peptides: importance of structure for activity. AB - Antimicrobial peptides are important components of innate immunity in species across the evolutionary scale. Unlike therapeutically used antibiotics, this class of peptides exert their activity by permeabilizing bacterial membranes. Despite the seemingly common mechanism of action, there is considerable variation in their primary structures, length and number of positive charges. Host-defense antimicrobial peptides have been the subject of extensive biophysical studies with a view at delineate structural requirements for activity. In this article, the structures of host defence antibacterial peptides and the structural requirements for activity are reviewed. PMID- 11945169 TI - Molecular diversity in gene-encoded, cationic antimicrobial polypeptides. AB - Gene-encoded, ribosomally synthesised antimicrobial peptides (AMPs) are an ancient and pervasive component of the innate defence mechanisms used by multicellular organisms to control the natural flora and combat pathogens. Bacteria also produce such AMPs to maintain ecological niches free of rival strains. Several hundred different peptides have been characterised to date, and they show a marked degree of variability in both sequence and structure, having evolved to act against distinct microbial targets in different physiological contexts. Many of these peptides appear to function via a selective, but not receptor-mediated, permeabilisation of microbial membranes, while others interact with specific membrane associated or intracellular targets. This review presents a broad survey of different amp structural classes, emphasising both their molecular diversity and underlying similarities. The mode of action of these peptides and potential for biomedical and other application is also briefly discussed. PMID- 11945170 TI - Pro-rich antimicrobial peptides from animals: structure, biological functions and mechanism of action. AB - Pro-rich antimicrobial peptides are a group of linear peptides of innate immunity isolated from mammals and invertebrates, and characterised by a high content of proline residues (up to 50%). Members of this group are predominantly active against Gram-negative bacterial species which they kill by a non-lytic mechanism, at variance with the majority of the known antimicrobial peptides. Evidence is accumulating that the Pro-rich peptides enter the cells without membrane lysis and, once in the cytoplasm, bind to, and inhibit the activity of specific molecular targets essential to bacterial growth, thereby causing cell death. This mode of action makes these peptides suitable for drug development efforts. In addition to antibacterial action, PR-39, one of the better characterised Pro-rich peptides from mammals, exerts other potentially exploitable biological activities, such as induction of syndecan expression in mesenchymal cells and inhibition of the NADPH oxidase activity of neutrophils, suggesting a role of this peptide in wound repair and inflammation. PR-39 also exerts a protective effect in various animal models of ischemia-reperfusion injury, preventing the post-ischemic oxidant production, and is a potent inducer of angiogenesis both in vitro and in vivo. Although the physiological relevance of all these effects has not yet been established, the above observations underscore the therapeutic potential of this peptide in a number of complex processes such as inflammation, wound repair, ischemia-reperfusion injury, and angiogenesis. PMID- 11945171 TI - Cathelicidin peptides as candidates for a novel class of antimicrobials. AB - Cathelicidin peptides are a numerous group of mammalian cationic antimicrobial peptides. Despite a common evolutionary origin of their genes, peptides display a remarkable variety of sizes, sequences and structures. Their spectra of antimicrobial activity are varied and cover a range of organisms that includes bacteria, fungi and enveloped viruses. In addition, they bind to and neutralize the effects of endotoxin. These features make this family of peptides good candidates in view of a therapeutic use. The most promising ones are currently under evaluation as leads for the development of novel anti-infectives, and synthetic variants are in an advanced stage of development for specific clinical applications. This review focuses on recent studies on the structure and in vitro and in vivo biological activities of these peptides. PMID- 11945172 TI - Development of short antimicrobial peptides derived from host defense peptides or by combinatorial libraries. AB - Recent increase of antibiotic-resistant pathogens demands exploration of novel antimicrobial molecules with unexploited mechanisms. Several hundred host defense peptides have been isolated from natural sources and their functions characterized. As host defense peptides have several advantages over classic antibiotics for resistant pathogens, there are many efforts to develop host defense peptides as therapeutic agents. In this review, focusing on the development of short antimicrobial peptides (< or = 18-mer), several examples are introduced that identify the active fragment from cyclic host peptides, or novel antimicrobial peptides derived from combinatorial libraries. Moreover, structure activity relationships of short antimicrobial peptides are discussed, and several methods for improving bioavailability as well as specificity of the peptides, such as D-amino acid replacements, unnatural amino acid replacements, and backbone modifications, are discussed. PMID- 11945173 TI - Multiple activities in lantibiotics--models for the design of novel antibiotics? AB - Lantibiotics are antibiotic peptides distinguished by the presence of the rare thioether amino acids lanthionine and/or methyllanthionine. They are produced by Gram-positive bacteria as gene-encoded precursor peptides and undergo post translational modification to generate the mature peptide. The structural gene for the prepeptide and the genes involved in biosynthesis, processing, export as well as regulation and producer strain self-protection are organized in clusters. Based on their structural and functional features lantibiotics are currently divided into two major groups. The flexible amphiphilic type-A lantibiotics act primarily by pore formation in the bacterial membrane, a mechanism which was recently shown, e.g. for nisin and epidermin, to involve the interaction with specific docking molecules such as the membrane precursor lipid II. The rather rigid and globular type-B lantibiotics inhibit enzyme functions through interaction with the respective substrates: mersacidin and actagardine inhibit the cell wall biosynthesis by complexing lipid II, whereas the cinnamycin-like peptides inhibit phospholipases by binding phosphoethanolamine. Lantibiotics have attracted much attention in recent years and undergone extensive characterization. New insights into the mode of action and structure-function relationships as well as the biochemistry and the genetics will be outlined in this review. PMID- 11945175 TI - Assembly of the Drosophila 26 S proteasome is accompanied by extensive subunit rearrangements. AB - The subunit contacts in the regulatory complex of the Drosophila 26 S proteasome were studied through the cross-linking of closely spaced subunits of the complex, and analysis of the cross-linking pattern in an immunoblot assay with the use of subunit-specific monoclonal antibodies. The cross-linking pattern of the purified 26 S proteasome exhibits significant differences as compared with that of the purified free regulatory complex. It is shown that the observed differences are due to extensive rearrangement of the subunit contacts accompanying the assembly of the 26 S proteasome from the regulatory complex and the 20 S proteasome. Cross linking studies and electron microscopic examinations revealed that these changes are reversible and follow the assembly or the disassembly of the 26 S proteasome. Although the majority of the changes observed in the subunit contacts affected the hexameric ring of the ATPase subunits, the alterations extended over the whole of the regulatory complex, affecting subunit contacts even in the lid subcomplex. Changes in the subunit contacts, similar to those in the regulatory complex, were detected in the 20 S proteasome. These observations indicate that the assembly of the 26 S proteasome is not simply a passive docking of two rigid subcomplexes. In the course of the assembly, the interacting subcomplexes mutually rearrange their structures so as to create the optimal conformation required for the assembly and the proper functioning of the 26 S proteasome. PMID- 11945174 TI - Iron chelation-induced senescence-like growth arrest in hepatocyte cell lines: association of transforming growth factor beta1 (TGF-beta1)-mediated p27Kip1 expression. AB - Iron is essential for cellular proliferation in all organisms. When deprived of iron, the growth of cells is invariably inhibited. However, the mechanism involved remains largely unclear. In the present study, we have observed that subcytotoxic concentrations of desferroxamine mesylate (DFO), an iron chelator, specifically inhibited the transition from G1 to S-phase of Chang cells, a hepatocyte cell line. This was accompanied by the appearance of senescent biomarkers, such as enlarged and flattened cell morphology, senescence-associated beta-galactosidase activity and reduced expression of poly(ADP-ribose) polymerase. Concomitantly, p27Kip1 (where Kip is kinase-inhibitory protein) was induced markedly, whereas other negative cell-cycle regulators, such as p21Cip1 (where Cip is cyclin-dependent kinase-interacting protein), p15INK4B and p16INK4A (where INK is inhibitors of cyclin-dependent kinase 4), were not, implying its association in the G1 arrest. Furthermore, the induction of p27Kip1 was accompanied by an increased level of transforming growth factor beta1 (TGF-beta1) mRNA. When neutralized with an anti-(TGF-beta1) antibody, p27Kip1 induction was completely abolished, indicating that TGF-beta1 is the major inducer of p27Kip1. Finally, DFO-induced senescence-like arrest was found to be independent of p53, since cell-cycle arrest was still observed with two p53-negative cell lines, Huh7 and Hep3B cells. In conclusion, DFO induced senescence-like G1 arrest in hepatocyte cell lines and this was associated with the induction of p27Kip1 through TGF-beta1, but was independent of p53. PMID- 11945176 TI - Transcriptional regulation of the human carboxyl ester lipase gene in THP-1 monocytes: an E-box required for activation binds upstream stimulatory factors 1 and 2. AB - The bile salt-stimulated carboxyl ester lipase (CEL) is important for the digestion and absorption of dietary lipids, and is expressed at high levels by the exocrine pancreas and the lactating mammary gland. However, the presence of CEL in human plasma suggests that the role of CEL in lipid metabolism may stretch beyond its function in the intestinal lumen, and possibly include interactions with cholesterol and oxidized lipoproteins to modulate the progression of atherosclerosis. We have used the CEL-expressing human monocytic cell line THP-1 to investigate the transcriptional regulation of the human CEL in monocytes. Analyses of the promoter region revealed that an E-box located at -47/-52 is necessary for CEL expression. Point mutations in the E-box almost completely abolish the transcriptional activity. Electrophoretic mobility-shift assay analyses reveal that the E-box binds the upstream stimulatory factors 1 and 2, and the binding of an upstream stimulatory factor-containing complex in THP-1 cells also requires the presence of a putative nuclear receptor-binding site at 60/-66. Furthermore, we demonstrate that the E-box is also necessary for CEL expression in the pancreas and the mammary gland, although there are tissue specific requirements for additional activating elements. PMID- 11945177 TI - Conserved hydrophobic amino acid residues in the N-terminal region of DnaA protein are involved in DnaA-DnaA interaction. AB - We previously reported that a leucine-zipper-like structure (I26, L33 and L40) located in the N-terminal region of DnaA is essential for the duplex opening at oriC by DnaA. In this study, we focused on three other conserved hydrophobic amino acid residues, L3, L10 and L17, and examined the function of DnaA proteins mutated in these amino acid residues. DnaA427 (L17S) and DnaA413 (L3S, L10S and L17S) were inactive for oriC DNA replication both in vitro and in vivo. Although these mutant DnaA proteins maintained their binding activities for both ATP and oriC, they were unable to induce the opening of duplex DNA at oriC. Glutathione-S transferase (GST)-fused wild-type DnaA interacted with wild-type DnaA but not with DnaA427 and DnaA413. Based on these results, we propose that conserved hydrophobic amino acid residues in the N-terminal region of DnaA are involved in DnaA oligomerization, in which DnaA-DnaA interaction is required. PMID- 11945178 TI - Evaluation of amplified rDNA restriction analysis (ARDRA) for the identification of cultured mycobacteria in a diagnostic laboratory. AB - BACKGROUND: The development of DNA amplification for the direct detection of M. tuberculosis from clinical samples has been a major goal of clinical microbiology during the last ten years. However, the limited sensitivity of most DNA amplification techniques restricts their use to smear positive samples. On the other hand, the development of automated liquid culture has increased the speed and sensitivity of cultivation of mycobacteria. We have opted to combine automated culture with rapid genotypic identification (ARDRA: amplified rDNA restriction analysis) for the detection resp. identification of all mycobacterial species at once, instead of attempting direct PCR based detection from clinical samples of M. tuberculosis only. RESULTS: During 1998-2000 a total of approx. 3500 clinical samples was screened for the presence of M. tuberculosis. Of the 151 culture positive samples, 61 were M. tuberculosis culture positive. Of the 30 smear positive samples, 26 were M. tuberculosis positive. All but three of these 151 mycobacterial isolates could be identified with ARDRA within on average 36 hours. The three isolates that could not be identified belonged to rare species not yet included in our ARDRA fingerprint library or were isolates with an aberrant pattern. CONCLUSIONS: In our hands, automated culture in combination with ARDRA provides with accurate, practically applicable, wide range identification of mycobacterial species. The existing identification library covers most species, and can be easily updated when new species are studied or described. The drawback is that ARDRA is culture-dependent, since automated culture of M. tuberculosis takes on average 16.7 days (range 6 to 29 days). However, culture is needed after all to assess the antibiotic susceptibility of the strains. PMID- 11945180 TI - Systematic quantitative overviews of the literature to determine the value of diagnostic tests for predicting acute appendicitis: study protocol. AB - BACKGROUND: Suspected acute appendicitis is the most frequent cause for emergency operations in visceral surgery worldwide. In approximately twenty percent of all cases however, the diagnosis is incorrect and patients undergo surgery without having acute appendicitis. Operations of bland appendices put patients at risk and entail a serious waste of resources. Several highly accurate tests have been introduced to diagnose acute appendicitis. The false positive rate however, has not changed over the last twenty years. Given the variation that exists in both practice and research, the uncertainty regarding the quality of the underlying evidence, there is a clear need for comprehensive, systematic and quantitative overviews of the diagnostic value of the various tests purported to be predictive of acute appendicitis. METHODS: Literature will be identified searching general bibliographic databases (MEDLINE and EMBASE), specialist computer databases (DARE, Cochrane Database of Systematic Reviews, conference proceedings, MEDION, SCISEARCH, BIOSIS) without language restrictions. We will contact experts and the manufacturers of tests. Hand-searching will complete our searches. Identified articles will be selected according to populations, tests, outcomes and study design. Papers meeting the selection criteria will be appraised to rate their methodological quality. Analysis will include exploration of heterogeneity in results. We will conduct meta-analyses to generate summary estimates of test accuracy measures and summary ROC curves where appropriate. If meta-analysis is considered to be inappropriate, we will describe the identified evidence in the context of appraised quality. DISCUSSION: These reviews should lead to formulation of recommendations for current practice and future research. PMID- 11945179 TI - Methylation and silencing of the retinoic acid receptor-beta 2 gene in cervical cancer. AB - BACKGROUND: Expression of the retinoic acid receptor beta2 (RAR-beta2), a putative tumor suppressor gene, is reduced in various human cancers, including squamous cell carcinomas (SCC) of the uterine cervix. The mechanism of the inhibition of RAR-beta2 expression remains obscure. We examined whether methylation of RAR-beta2 gene could be responsible for this silencing in cervical SCC. METHODS: Expression of RAR-beta2 mRNA and methylation status of the 5' region of RAR-beta2 gene were examined in 20 matched specimens from patients with cervical SCC and in three cervical cancer cell lines by Northern blot analysis and methylation-specific PCR (MSP) assay or Southern blot analysis respectively. RESULTS: In 8 out 20 cervical SCC (40%) the levels of RAR-beta2 mRNA were decreased or undetectable in comparison with non-neoplastic cervix tissues. All 8 tumors with reduced levels of RAR-beta2 mRNA expression showed methylation of the promoter and the first exon expressed in the RAR-beta2 transcript. The RAR-beta2 gene from non-neoplastic cervical tissues was mostly unmethylated and expressed, but methylated alleles of the gene were found in three samples of the morphologically normal tissues adjacent to the tumors. Three cervical cancer cell lines with extremely low level of RAR-beta2 mRNA expression, SiHA, HeLA and CaSki, also showed methylation of this region of the RAR-beta2 gene. CONCLUSIONS: These findings suggest that methylation of the 5' region of RAR-beta2 gene may contribute to gene silencing and that methylation of this region may be an important and early event in cervical carcinogenesis. These findings may be useful to make retinoids more effective as preventive and therapeutic agents in combination with inhibitors of DNA methylation. PMID- 11945181 TI - The families evaluation on management, care and disclosure for terminal stage cancer patients. AB - BACKGROUND: Quality of life is an important concept which is subjective and personal; what is an acceptable quality of life to one may be 'worse than death' to another. The objective of this study was to develop and validate a questionnaire to assess relatives' perceptions and attitudes towards their terminal stage cancer patients' management (information disclosure, treatment choice, hospitalization and support-communication and care) including aspects regarding end-of-life and quality-of-life decisions. METHODS: The final study consisted of 146 relatives of advanced terminal stage cancer patients receiving palliation, attending a Pain Relief and Palliative Care Unit. The questionnaire incorporated 6 multi-item and 7 single-item scales, and was developed following a systematic review of measures appropriate for use in palliative care settings. RESULTS: Following analysis of the 25-item scale, the questionnaire has been validated as a shortened 21-item scale consisting of 5 multi-item and 5 single item scales. Factor analysis was based upon information disclosure, hospitalization, and support-communication demonstrating Cronbach's alpha coefficients of 0.66, 0.5 and 0.70 respectively. Average item totals and inter item scale correlations were between 0.62-0.70, with convergent validity correlations between 0.60-0.86. The questionnaire was well accepted by all subjects with an 8-10 minute completion time. CONCLUSION: The shortened 21-item self-assessment questionnaire may provide acceptable and valid assessment of caregiver(s)/Greek cancer patients' relatives perceptions on palliative care. PMID- 11945183 TI - Comparison of duration of deafness and tumour invasion to the inner ear from metastatic tumours of the internal auditory canal: human temporal bone pathology. AB - Four cases (seven ears) of metastatic tumour of the internal auditory canal were studied. The histopathological findings confirmed that the inner ear invasion of the tumour follows a unique course, as reported in the literature. Relationship between duration of deafness and extent of tumour invasion in the inner ear is discussed. It is suggested that the deafness could occur via neural invasion or compression near the ductus spiralis foraminosus. PMID- 11945184 TI - Effect of anti-fungal nasal lavage with amphotericin B on nasal polyposis. AB - Recent studies have suggested that allergic fungal rhino-sinusitis could be involved in the development of nasal polyposis. The aim of this study was to evaluate the response of anti-fungal nasal lavages. Patients performed nasal lavage with 20 ml of a one per one thousand amphotericin B suspension in each nostril, twice a day, for four weeks. In addition, all patients continued their saline nasal lavage and their conventional topical corticosteroid spray. This study included 74 patients, with a mean age of 46 years (range from 19 to 73). Before anti-fungal treatment, the distribution of nasal polyposis, according to Malm, was: 13 patients in stage I (17.5 per cent), 48 patients in stage II (65 per cent) and 13 patients in stage III (17.5 per cent). After anti-fungal nasal lavages, the total disappearance of nasal polyposis was observed in 29 patients (39 per cent). Eight patients were stage I, 21 stage II, and none stage III. In patients who have had previous endoscopic polypectomy and functional endoscopic sinus surgery, total disappearance of nasal polyposis was seen in 24 patients (47 per cent). Hyper-reactivity to fungal organisms could be one of the mechanisms underlying the development of nasal polyposis. A direct effect of amphotericin B suspension on the integrity of the cell membrane of the polyps' epithelium could not be excluded. PMID- 11945185 TI - Immunocytochemical localization of lysozyme and lactoferrin attached to surface bacteria of the palatine tonsils during infectious mononucleosis. AB - Bacterial samples were obtained from the tonsillar surfaces of seven patients (four males, three females; median age 18 years, range 15 to 21 years) suffering from acute infectious mononucleosis with concomitant pharyngotonsillitis, and from five healthy controls. By using gold-labelled antiserum to human lysozyme and lactoferrin, micro-organisms on the tonsillar surfaces coated with these antibacterial substances could be identified by tracing the gold particles in the transmission electron microscope. In healthy individuals, most of the bacteria were coated with lysozyme and significantly more bacteria were coated with lysozyme than with lactoferrin (p < 0.01). In patients there was a non significant reduction in lysozyme-coating of the bacteria, whereas lactoferrin coating was significantly increased (p < 0.01). Changes in the lysozyme and/or lactoferrin coating of the tonsillar surface bacteria on the palatine tonsils during infectious mononucleosis cannot explain the tendency to immense local bacterial colonization with commensals and proneness to bacterial penetration into the epithelial cells. PMID- 11945186 TI - The cause of death in patients with head and neck squamous cell carcinoma. AB - For patients with head and neck squamous cell carcinoma the cause of death is not well described in world literature. We report data on 106 patients diagnosed with head and neck squamous cell carcinoma who subsequently died. The literature related to this topic is discussed, and recommendations are made for data collection. PMID- 11945187 TI - Accessory nerve monitoring and stimulation during neck surgery. AB - Iatrogenic injury to the spinal accessory nerve following surgical procdures in the neck is well recognized in causing significant morbidity to patients, with shoulder pain and loss of function being particularly problematic. We have used a Magstim Neurosign 100 peripheral nerve monitor, that is most often used in our practice to monitor the facial nerve during middle ear and parotid surgery, to monitor the accessory nerve during neck surgery. Ten patients undergoing accessory nerve-sparing neck dissection, or excision biopsy of neck mass had their accessory nerve monitored during the procedure. No patient suffered injury of the nerve. In several cases the nerve closely adhered to the tissue being resected, and in two cases, the nerve bifurcated or gave off branches. We found that the monitor aided identification and preservation of the nerve. PMID- 11945188 TI - Waiting times during the management of head and neck tumours. AB - The waiting times incurred during the management of 75 consecutive head and neck oncology patients attending for post-treatment follow-up were reviewed. Data were gleaned from general practitioner (GP) referral letters, patient case-notes as well as radiology and histology reports. The mean time for GP referral to ENT was 5.1 weeks. From ENT to endoscopy was 3.1 weeks, to histology 3.5 weeks, to computed tomography (CT) scan 5.6 weeks, to magnetic resonance scan (MR) 4.1 weeks, to primary radiotherapy 10.3 weeks and to surgery 5.5 weeks. The mean symptom duration prior to referral was 4.9 months. Our results compare unfavourably with the standards recommended by the BAO-HNS. Local modifiations may improve matters, but significant increases in funding, manpower and equipment are required to achieve the stipulated standards. Moreover, criteria for referral have to be re-emphasized and patient education has to be addressed as these appear to contribute the longest delay in the diagnosis of head and neck tumours. PMID- 11945189 TI - Fever and acquired torticollis in hospitalized children. AB - Acute torticollis due to non-traumatic atlanto-axial subluxation (AAS) is often seen in children presenting with inflammatory conditions of the upper respiratory tract and the neck. Grisel's syndrome is the eponym given to this condition. These patients may present earlier in the disease process without evident subluxation. Thus, early recognition of the condition with prompt commencement of appropriate conservative treatment could halt the progression into Grisel's syndrome. The purpose of this study is to address the importance of early recognition of inflammatory torticollis that can be treated successfully by conservative methods. A retrospective review was made of the case files and radiological investigations of 13 children with fever and torticollis who were treated in the neurosurgery unit of Hamad General Hospital in Qatar, over a two year period from July 1996 to July 1998. The children were aged between three and 12 years with a male to female ratio of 3:1. All patients arrived at the hospital within 48 hours of onset of torticollis and almost all had manifestations of upper respiratory tract or head and neck infections. Radiological examination by cervical spine X-rays, computerized tomography (CT) or magnetic resonance images (MRI) revealed that only three cases out of 13 had AAS. All patients underwent conservative treatment that included rest, neck collar, simple analgesics and antibiotics, where appropriate. A muscle relaxant was used in nine cases and Halter traction was applied to the three with AAS. All patients responded well to treatment and none required surgical intervention for AAS. We conclude that the majority of children presenting acutely with inflammatory torticollis have rotational deformity only without AAS. Progression to the latter, i.e. Grisel's syndrome, may be aborted should the diagnosis be made early and conservative treatment initiated in time. On the other hand, delay in diagnosis would deprive these children an opportunity of receiving effective conservative treatment. PMID- 11945190 TI - Parotid gland surgery: a retrospective review of 108 cases. AB - One hundred and eight parotidectomies performed by a single consultant were reviewed. Eighty-five patients had primary parotid disease, 23 patients had extra parotid primaries. Pleomorphic adenoma was the most common histological diagnosis. In patients with primary parotid disease, a post-operative temporary facial nerve palsy was noted in 15 patients, with a further four developing a permanent palsy. Patients with metastatic disease to the parotid had a poor prognosis. PMID- 11945191 TI - Pneumocystis carinii infection in bilateral aural polyps in a human immunodeficiency virus-positive patient. AB - Pneumocystis carinii is an opportunistic infection found in patients with impaired immunity. Under favourable conditions the parasite can spread via the blood stream or lymphatic vessels and cause extrapulmonary dissemination. We report a case of P carinii infection presenting as bilateral aural polyps, otitis media and mastoiditis in human immunodeficiency (HIV)-positive patient with no history of prior or concomitant P carinii infection. PMID- 11945192 TI - Sudden hearing loss in chronic myelogenous leukaemia implicating the hyperviscosity syndrome. AB - Sudden sensorineural hearing loss that presents as the initial sign of haematological disease is very rare. Chronic myelogenous leukaemia has been implicated as a causative factor of sudden sensorineural hearing loss. A 49-year old male presented with unilateral sudden sensorineural hearing loss. The patient was found to have chronic myelogenous leukaemia during a work-up for his hearing loss. We present a case of a chronic myelogenous leukaemia patient whose first manifestation was sudden sensorineural hearing loss. We presume that cochlear vessel occlusion as a result of elevated blood viscosity was responsible for this patient's hearing loss. Early onset of sudden deafness in a chronic myelogenous leukaemia patient may be due to the hyperviscosity syndrome and it may be possible to reverse hearing loss through early leukapheresis. PMID- 11945193 TI - An unusual cause of epistaxis: a haemophilic pseudotumour in a non-haemophiliac, arising in a paranasal sinus. AB - Most cases of epistaxis are due to simple causes and are easily treated on an out patient basis. However, there are some cases where the origin of bleeding is not obvious or arises from an unusual pathological source. The authors describe a case of epistaxis due to a mass in the maxillary antrum that when biopsied showed the histological appearances of a haemophilic pseudotumour. The patient was anticoagulated on warfarin for a cardiac valve replacement and this was thought to be the cause of the ongoing haemorrhage necessary for development of the pseudotumour. Even in haemophiliacs, pseudotumours are rare and we believe this case is unique in that the patient is a non-haemophiliac. The epistaxis was eventually controlled by external beam radiotherapy to the pseudotumour. The management of this case is outlined as well as a review of the literature on haemophilic pseudotumour. PMID- 11945194 TI - Haemangioma of the frontal sinus. AB - A 71-year-old lady underwent successful excision of a haemangioma of the frontal sinus, via an osteoplastic flap approach. Haemangioma of the paranasal sinuses is an extreme rarity. A case is presented and the literature reviewed. PMID- 11945195 TI - An aberrant artery as a cause of massive bleeding following adenoidectomy. AB - Bleeding following adenoidectomy is a rare complication of this surgical procedure, and usually occurs in the first post-operative hours. In almost all cases it is related to adenoid remnants. Removal of these remnants under a second general anaesthetic is the treatment of choice. We report a case of massive bleeding three days following adenoidectomy related to the injury of an aberrant ascending pharyngeal artery during adenoidectomy performed elsewhere. Signs of manifest shock necessitated an immediate ligature of the external carotid artery and its individual branches. The post-operative course was uneventful. Two years later the boy underwent tonsillectomy without complications. PMID- 11945196 TI - Primary nasopharyngeal tuberculosis in a patient with the complaint of snoring. AB - Isolated nasopharyngeal tuberculosis is a rare condition, even in endemic tuberculosis areas. The most common presentation of nasopharyngeal tuberculosis is with a cervical lymphadenopathy followed by nasal discharge or obstruction. Here we present a 58-year-old patient with nasopharyngeal tuberculosis whose only complaint was snoring. Her oropharyngeal and anterior rhinoscopic examination was normal. On endoscopic examination, mucosal oedema and hyperaemia of the nasopharynx was observed. There was no cervical lymphadenopathy. The tuberculin skin test was positive and histopathological examination of the biopsy taken from posterior nasopharyngeal wall supported the diagnosis of tuberculosis. After anti tuberculosis therapy, the snoring stopped and the nasopharyngeal examination was normal. PMID- 11945197 TI - Primary aspergillosis of the larynx associated with CD4+ T lymphocytopenia. AB - The purpose of this study was to report the first case (to our knowlege) of primary laryngeal aspergillosis in a patient with underlying CD4(+) T lymphocytopenia. Laryngeal involvement of Aspergillus is more commonly seen as a part of a wider infection involving the respiratory system in an immunocompromised host. However, primary infection of the larynx is extremely rare. Although there were 12 cases of primary laryngeal aspergillosis previously reported in healthy subjects, there is no known study describing immunological findings in detail. We report a case of primary laryngeal aspergillosis in a healthy 79-year-old male. The examination of his immunity subsequently revealed that there was a marked decline in the number of CD4(+) T lymphocytes and a decrease in the ratio of CD4(+) to CD8(+). It is suggested that it is essential to examine the defence mechanisms, specifically cell-mediated immunity in a patient showing primary laryngeal aspergillosis. PMID- 11945198 TI - Impacted foreign body in the hypopharynx and oesophagus. AB - We report a rare case of an unusual impacted foreign body (titanium mesh stent) in the hypopharynx and upper oesophagus of a 66-year-old gentleman. He was a known patient with carcinoma of the mid-oesophagus admitted under the gastroenterologists for dilatation of the carcinomatous stricture with a stent. During the procedure the titanium mesh spring coil got displaced and was impacted in the retropharyngeal space, submucosally in the hypopharynx and upper oesophagus. He was referred to us for surgical removal of the stent, which he underwent successfully. We believe this to be the first case of such an unusual foreign body to be reported in the literature. PMID- 11945199 TI - Common carotid artery dissection: a rare cause for cervical pain. AB - Spontaneous dissections of extracranial carotid arteries occur most frequently in the internal carotid artery. In contrast, common carotid artery dissection (CCA) is a rare cause of cerebral ischaemia with only a few cases having been reported. We present the case of a 59-year-old male patient who was referred to our clinic with left cervical pain. The patient was otherwise asymptomatic. Magnetic resonance imaging (MRI) showed enhancement and double lumen in the left CCA without affecting the carotid bulb, internal carotid artery or aortic arch. We discuss the diagnostic and therapeutic management of the disease, focusing on its differentiation from other causes of cervical pain. PMID- 11945200 TI - A case of buccal abscess; originating from an ectopic accessory parotid gland? AB - We report a case of an abscess from an 'ectopic' accessory parotid gland in the cheek demonstrated by sialography and computed tomography (CT). The accessory parotid gland was ectopically located anterolateral to the masseter muscle and isolated from the main parotid gland. The orifice and ductal system of the ectopic accessory parotid gland were separated from those of Stensen's duct. The abscess developed from this ectopic accessory gland, and the main parotid gland was free of inflammation. Using sialography and CT, we confirmed the presence of this ectopic accessory gland. PMID- 11945201 TI - Focal myositis of the neck with idiopathic orbital myositis. AB - Focal myositis (of Heffner) in the right trapezius and paraspinal muscles accompanied by a pseudotumour of the left orbit has not been reported previously. The clinical, pathological and radiological features of these unusual benign pseudotumours of the head and neck are discussed. Computerized tomography (CT) scan of the neck and orbital ultrasound were suggestive of an inflammatory process without an abscess formation. This was followed by fine needle aspiration cytology (FNAC), which confirmed the diagnosis. The patient was treated with intravenous steroids and antibiotics, that led to complete resolution of symptoms, and there was no recurrence at six months follow-up. This report highlights the importance of imaging in inflammatory neck swellings. PMID- 11945202 TI - Respiratory epithelial adenomatoid hamartoma of the maxillary sinus. AB - A case of respiratory epithelial adenomatoid hamartoma of the maxillary sinus is reported. Glandular hamartomas involving the sinonasal tract have received only limited documentation in the literature. The differential diagnosis of adenomatoid hamartoma includes schneiderian papilloma of the inverted type and adenocarcinoma. Limited but complete surgical resection is the treatment of choice. PMID- 11945203 TI - Conversion of percutaneous tracheostomy-a practical solution. PMID- 11945204 TI - Concurrent use of antiretrovirals and anticonvulsants in a rural population. PMID- 11945205 TI - Eosinophilic granuloma of the femur in an HIV-1-positive patient. AB - A case of eosinophilic granuloma in the right femur of an HIV-1-infected patient is described, and the possible pathogenetic role of HIV infection in eosinophilic granuloma formation is discussed. PMID- 11945206 TI - AIDS-related primary brain lymphoma in Michigan, January 1990 to December 2000. AB - To examine the effect of highly active antiretroviral therapy (HAART) on the occurrence of primary brain lymphoma (PBL), determine the risk factors for PBL, and assess the difference in survival between individuals who did and did not develop PBL, data were analyzed from the Michigan Adult/Adolescent Spectrum of HIV Disease project (ASD). Among 4,026 HIV-infected individuals enrolled in ASD between January 1990 and December 2000, 64 (1.6%) were diagnosed with PBL. The incidence rate of PBL declined from 5.6 cases per 1000 person-year in the pre HAART era to 2.1 cases per 1000 person-year in post-HAART era. In the multivariate analysis, individuals whose CD4 count at entry in ASD was either <50 cells/microL (OR: 5.0) or 50-200 cells/microL (OR: 2.3) were significantly more likely to develop PBL than those with a CD4 count >/= 200 cells/microL. PBL was a terminal condition in these patients. The median survival time was consistently shorter among individuals who developed PBL than those who did not, regardless of their CD4 count at entry into ASD. PMID- 11945207 TI - Prospective study of emotional reactions to changes in HIV viral load. AB - Advances in antiretroviral therapies for HIV infection have given rise to new hope for the long-term survival of people living with HIV/AIDS. The current study examined changes in depression, anxiety, and HIV treatment attitudes that may co occur with improvements and setbacks in HIV disease progression in a prospective cohort of 166 HIV-positive men and women. Results from a 9-month prospective observation period showed that persons who experienced increases in viral load, as indexed by medical chart abstraction, reported modest increases in depression and decreases in positive attitudes toward HIV treatments. Persons who experienced substantive decreases in viral load to undetectable levels demonstrated meaningful reductions in depression, particularly affective symptoms of depression after statistically controlling for years living with HIV, HIV symptoms, and age. Results suggest that emotional distress and perceptions of HIV treatments change in parallel to markers of HIV disease progression, especially with regard to reduced emotional distress in response to clinically significant changes in viral load. PMID- 11945208 TI - Physical and sexual abuse in the lives of HIV-positive women enrolled in a primary medicine health maintenance organization. AB - Over the past several years, the proportion of all cases of AIDS in the United States among adult and adolescent women has more than tripled from 7% in 1985 to 23% in 1998. Information obtained in the present study suggests that care providers need to be aware of the unique life circumstances of women with AIDS, which may predispose them to a number of negative health and mental health outcomes. Charts of the first 100 women enrolled in an intensive home-based primary medical HMO for people with advanced HIV/AIDS were examined retrospectively for evidence of trauma. Results from a chart review and nursing care assessments of these patients revealed that women with HIV/AIDS were significantly more likely to have had traumatic life experiences compared to the larger population [as measured in a National Comorbidity Survey (NCS)]. In this study, one-half of the patients reported a lifetime history of sexual assault compared to 9% of the general population, one-third reported a history of incest compared to 12% in the NCS, and 83% reported significant physical abuse compared to 4% in the NCS. Such traumatic life experiences are frequently associated with high rates of psychiatric comorbidity, substance abuse, and possible nonadherence to health care. Providers of AIDS care need to be aware of the complex mental health and psychosocial needs of traumatized women with AIDS and make better use of collateral mental health providers and consultation. The ways in which this particular sample of women may be nonrepresentative of women living with AIDS, in general, including the observation that they may be a particularly traumatized and challenging cohort, and other limitations of the data, including the methods used for chart review, are discussed. PMID- 11945209 TI - HIV-positive women and minority patients' satisfaction with inpatient hospital care. AB - Although patient satisfaction has been examined in relation to HIV services for ambulatory and managed care, less is known about perceptions of hospital care, particularly for HIV-positive women and minorities. The purpose of this study was to examine HIV-positive women and minority patients' satisfaction with hospital care. The study was part of a larger funded study that explored potential health care disparities for HIV-positive women and minority persons in the era of HIV combination drug therapy. A convenience sample of 50 HIV-positive persons was recruited from four medical centers/hospitals in a South Florida community. The multi-ethnic sample included 31 women and 19 men. The survey tool used was Cleary's HIV-Infected Patient's In-Hospital Questionnaire. Findings revealed that participants were generally satisfied with their hospital care. Physicians, nurses, and the hospital environment received satisfactory ratings. However, several problem areas were identified, including pain management and education on side effects of HIV medications, indicating the need for interventions to improve care. Experimental AIDS drugs were discussed with less than half of the participants, suggesting that HIV-positive women and minority patients may not have equal access to clinical drug trials. Further research is also needed to determine whether attitudes conveyed by health care providers influence HIV positive patients to be wary of advance directives. The competence of nurses experienced in acute-care nursing of persons with HIV/AIDS was an important factor in patient satisfaction. A lack of experienced acute-care AIDS nurses may ultimately lead to a decrease in HIV-positive patients' satisfaction with hospital care. PMID- 11945210 TI - SMART trial begins. PMID- 11945211 TI - AIDS cases rising again. PMID- 11945213 TI - Addressing tobacco in managed care: documenting the challenges and potential for systems-level change. PMID- 11945214 TI - The feasibility of paper-based Tracking Codes and electronic medical record systems to monitor tobacco-use assessment and intervention in an Individual Practice Association (IPA) Model health maintenance organization (HMO). AB - Despite evidence of its effectiveness, tobacco cessation is not systematically addressed in routine healthcare settings. Its measurement is part of the problem. A pilot study was designed to develop and implement two different tobacco tracking systems in two independent primary care offices that participated in an IPA Model health maintenance organization in Portland, Oregon. The first clinic, which utilized a paper-based charting system, implemented CPT-like tracking codes to measure and report tobacco-cessation activities, which were eventually included in the managed-care organization's (MCO) claims database. The second clinic implemented an electronic tracking system based on its computerized electronic medical record (EMR) charting system. This paper describes the pilot study, including the processes involved in building provider acceptance for the new tracking systems in these two clinics, the barriers and successes encountered during implementation, and the resources expended by the clinics and by the MCO during the pilot. The findings from the 3-month implementation period were that documentation of tobacco-use status remained stable at 42-45% in the paper-based clinic and increased from 79% to 88% in the EMR clinic. This pilot study demonstrated that Tracking Codes are a feasible preventive-care tracking system in paper-based medical offices. However, high levels of effort and support are needed, and a critical mass of insurers and health plans would need to adopt Tracking Codes before widespread use could be expected. Results of the EMR-based tracking system are also reviewed and discussed. PMID- 11945216 TI - Smoking cessation in pregnancy: failure of an HMO pilot project to improve guideline implementation. AB - This pilot project investigated whether a system introduced by a health maintenance organization (HMO) could encourage obstetric providers to implement smoking-cessation guidelines in their offices. Staff from participating offices was trained in cessation counseling and paid $150 for each pregnant smoker counseled. Data were collected from chart review from participating physicians and a telephone survey of all HMO members giving birth before and after program implementation. Eighteen physician practices participated, representing 27 office sites and 80 physicians. Sixty-six staff members were trained in cessation counseling. Chart review revealed that identification of smoking status increased from 90% to 96% (p=0.03), but documentation of advice to quit worsened (62% vs. 24%, p=0.03). The HMO received claims for counseling from four pregnant smokers out of the 21 identified on chart review. The telephone survey revealed a similar prevalence of current smokers in the baseline and post-intervention samples (15% vs. 13%) but more former smokers (11% vs. 22%) in the follow-up. For the HMO as a whole, fewer smokers reported being advised to quit in the follow-up survey (86% vs. 65%). Few smokers reported being counseled to quit at baseline or follow-up (11% vs. 3%). None of the smokers who received obstetric care from a pilot group physician reported awareness of the smoking-cessation benefit despite the placement of brochures in participating offices advertising the program. This pilot project to reimburse for cessation counseling did not increase smoking cessation advice or counseling in participating physician's offices. Smoking cessation counseling worsened across the entire HMO obstetric community during the intervention. A more comprehensive systems approach and a larger HMO market share or a partnership among multiple HMOs may be required to increase smoking cessation guideline implementation. PMID- 11945215 TI - Pilot study of enhanced tobacco-cessation services coverage for low-income smokers. AB - This study explored the feasibility of covering nicotine replacement therapy (NRT) and paying for pharmacist-delivered smoking cessation counseling at the time of NRT pick-up for low-income, managed Medicaid and Basic Health Plan (a state insurance program) enrollees. A prospective pilot intervention was used at two community health centers (CHCs) and two community pharmacies. Participants were adult managed-Medicaid or Basic Health Plan enrollees who attended the pilot CHCs and smoked. An innovative insurance benefit that included coverage for NRT and $15 payment to the pharmacist to deliver cessation counseling with each prescription fill. Proportion of eligible patients who used the cessation benefit and patient and pharmacist satisfaction with the intervention. During the 9-month intervention, 32 patients at the pilot clinics were referred for NRT and pharmacist-delivered counseling. This number represented roughly 5% of eligible smokers. Of these, 26 received NRT with concomitant pharmacist-delivered cessation counseling at least once. Recipients reported a high level of satisfaction with this intervention. Pharmacists indicated they would continue providing counseling if reimbursement remained adequate and if counseling lasted no longer than 5-10 min. However, 12 (38%) who were referred were no longer insured by the sponsoring plan by the end of the 9-month pilot period. Pharmacist delivered cessation counseling may be feasible and merits further study. More importantly, this pilot reveals two key obstacles in our low-income, culturally diverse setting: low participation and rapid turnover of insureds. Future interventions will need to address these barriers. PMID- 11945217 TI - Capturing tobacco status using an automated billing system: steps toward a tobacco registry. AB - In 1999, Group Health Cooperative (GHC), a health system with 575,000 enrollees, launched a quality improvement initiative to systematically record patient tobacco-use status and provider intervention using an automated billing system. Performance feedback and senior-level incentives were added to foster compliance with the automated recording. Prior to this period, tobacco-use status was recorded primarily via a paper-based chart system, with billing-system recording averaging only 7.5% of primary care visits. In 2000, tobacco-use status was recorded using the billing system in an average of 82% of visits (p<0.001). Significant increases (p<0.0001) were also observed for the absolute number of visits in which automated entry of tobacco-use status was documented using the billing system, as well as for visits where intervention was documented. In 1998, tobacco use was documented in 22,086 visits, with intervention documentation in 13,235 of these visits. By 2000, tobacco-use documentation increased to 76,180, with intervention documentation in 45,527. This work demonstrates the feasibility of using automated performance feedback and senior-level incentives to increase provider compliance with a new system of tobacco status identification and intervention. Other potential uses of this system, and potential limitations, are discussed. PMID- 11945218 TI - Academic profiling of tobacco-related performance measures in primary care. AB - Academic detailing and data feedback are two methods that have been used to change provider behavior. Academic profiling is proposed as an intervention that combines provider educational outreach and peer-comparison feedback of data generated from chart reviews and health plans. This project assessed the feasibility of academic profiling, using baseline measures to assess provider performance in identifying and treating patients who smoke. The pilot study was undertaken with four primary care practices in Maine. Two health plans shared administrative claims data on adult patients of participating providers. Two educational sessions were conducted: one including feedback of tobacco-related chart documentation and claims for nicotine replacement and bupropion (Zyban), and the other, coding for tobacco use (ICD-9 305.1) in adults enrolled in two health plans during 1998. A mailed survey assessed provider attitudes following the intervention. Among 24 providers, 80% attended the first session and 70% attended the second session. Provider documentation of tobacco status in the medical records varied from 68% to 100%. The frequency of tobacco pharmacotherapy claims for adult health plan enrollees having a provider visit in 1998 varied from 0% to 4.6% (mean 1.5%) by provider. The frequency of tobacco use diagnosis claims (ICD-9 305.1) varied from 0% to 19.8% by provider. More than 90% of the providers who reviewed the profiling graphs found the data were understandable, and 66% reported that the sessions helped them improve the ways they interact with patients who smoke. Practices vary in tobacco-related documentation, the prescribing of tobacco pharmacotherapy, and the coding for tobacco use. Providers are willing to participate in educational outreach using peer-comparison feedback, presenting opportunities to improve performance in the treatment of tobacco dependence. PMID- 11945219 TI - Somatic cell cloning in Buffalo (Bubalus bubalis): effects of interspecies cytoplasmic recipients and activation procedures. AB - Successful nuclear transfer (NT) of somatic cell nuclei from various mammalian species to enucleated bovine oocytes provides a universal cytoplast for NT in endangered or extinct species. Buffalo fetal fibroblasts were isolated from a day 40 fetus and were synchronized in presumptive G(0) by serum deprivation. Buffalo and bovine oocytes from abattoir ovaries were matured in vitro and enucleated at 22 h. In the first experiment, we compared the ability of buffalo and bovine oocyte cytoplasm to support in vitro development of NT embryos produced by buffalo fetal fibroblasts as donor nuclei. There were no significant differences (p > 0.05) between the NT embryos derived from buffalo and bovine oocytes, in fusion (74% versus 71%) and cleavage (77% versus 75%) rates, respectively. No significant differences were also observed in blastocyst development (39% versus 33%) and the mean cell numbers of day 7 cloned blastocysts (88.5 +/- 25.7 versus 51.7 +/- 5.4). In the second experiment, we evaluated the effects of activation with calcium ionophore A23187 on development of NT embryos after electrical fusion. A significantly higher (p < 0.05) percentage of blastocyst development was observed in the NT embryos activated by calcium ionophore and 6-DMAP when compared with 6-DMAP alone (33% versus 17%). The results indicate that the somatic nuclei from buffalo can be reprogrammed after transfer to enucleated bovine oocytes, resulting in the production of cloned buffalo blastocysts similar to those transferred into buffalo oocytes. Calcium ionophore used in conjunction with 6-DMAP effectively induces NT embryo development. PMID- 11945220 TI - Pathways involved in RGD-mediated calcium transients in mature bovine oocytes. AB - An arginine-glycine-aspartic acid (RGD)-containing peptide has been reported to generate calcium transients in bovine oocytes similar to those observed at fertilization. The research objective herein was to evaluate the response of bovine oocytes to an RGD peptide after injection with known antagonists of calcium releasing mechanisms in order to determine the initial calcium releasing pathway. Oocytes were injected with either heparin, an inhibitor of inositol 1,4,5-trisphosphate (IP3) induced calcium response, or procaine, which inhibits calcium release through the ryanodine receptor. Oocytes injected with heparin prior to RGD exposure did not display a calcium response. Oocytes injected with procaine prior to RGD exposure did exhibit a calcium response. Electroporation of IP3, caffeine, or exposure to RGD alone elicited a calcium response for each treatment group. Injection of heparin, procaine, vehicle medium (VM), or exposure to a non-RGD-containing peptide alone failed to elicit a calcium response. The data indicates that the RGD peptide is able to induce calcium transients in oocytes inhibited with procaine, but not those inhibited with heparin. These data suggest the pathway whereby the RGD peptide induces the first intracellular calcium transient in bovine oocytes is through IP3-mediated stores. PMID- 11945221 TI - Glucose and pyruvate metabolism of preimplantation goat blastocysts following in vitro fertilization and parthenogenetic activation. AB - The energy metabolism of preimplantation embryos can be used to predict viability and postimplantation development. Although preimplantation development and mean blastocyst cell numbers of goat in vitro-fertilized (IVF) embryos and chemically activated parthenogenotes are comparable, mammalian parthenogenotes are not viable, with most dying shortly after implantation. The objective of this study was to compare glucose and pyruvate metabolism of IVF goat blastocysts with that of parthenogenetic blastocysts developing from chemically activated oocytes. Embryos derived from IVF and parthenogenotes produced by exposing oocytes to either ionomycin or ethanol followed by 6-dimethylaminopurine (6-DMAP) were cultured in G1.2/G2.2 sequential culture media. Metabolism was determined for individual blastocysts using [5-3H]glucose and [2-14C]pyruvate to determine glycolytic and Kreb's cycle activity, respectively. Data were analyzed by ANOVA. A significantly higher percentage of activated oocytes underwent cleavage and developed to the blastocyst stage compared to IVF oocytes (p < 0.05). There was no significant difference in glucose or pyruvate metabolism between IVF and parthenogenetically activated blastocysts. Mean glucose metabolism through glycolysis was 154.9 +/- 29.1, 130.3 +/- 17.1, and 129 +/- 16.5 pmol/embryo/3 h for IVF, ethanol-activated, and ionomycin-activated blastocysts, respectively. Mean pyruvate metabolism through the Kreb's cycle was 28.1 +/- 8.0, 15.8 +/- 4.2, and 24.4 +/- 4.4 in pmol/embryo/3 h for IVF, ethanol-activated, and ionomycin activated blastocysts, respectively. Our results suggest that known differences in postimplantation development observed in IVF versus parthenogenetic embryos cannot be attributed to differences in pyruvate or glucose metabolism in the preimplantation blastocysts. Thus, these activation protocols result in embryos capable of appropriate regulation of key metabolic enzymes. PMID- 11945222 TI - Cell synchronization for the purposes of nuclear transfer in the bovine. AB - We have examined the reprogramming ability of donor fibroblast nuclei in various phases of the cell cycle, upon transfer to cytoplasts, using a bovine nuclear transfer (NT) model. Bovine fetal fibroblasts were cultured in reduced serum and conditioned medium to induce quiescence (G0) and treated with nocodazole to induce M phase arrest. Unsynchronized actively dividing cells (control) were mainly in G1. Cells synchronized in G0, M, and G1 phase were transferred to enucleated bovine MII oocytes by direct injection using the Piezo-Drill microinjector. NT oocytes were artificially activated following injection. Cells at the M phase were also transferred to enucleated oocytes after artificial activation. Cells induced into quiescence by serum starvation and unsynchronized donor cells produced the highest rates of development to the morula/blastocyst stage (20% and 18%, respectively). Development to blastocyst was significantly higher in parthenogenetic controls compared to NT embryos. The transfer of M phase nuclei to MII cytoplasts was not associated with high development to the blastocyst stage. Nevertheless, determining the viability of these embryos requires transfer to recipient animals and assessment of in vivo development. PMID- 11945223 TI - Simplification of bovine somatic cell nuclear transfer by application of a zona free manipulation technique. AB - Contemporary nuclear transfer techniques often require the involvement of skilled personnel and extended periods of micromanipulation. Here, we present details of the development of a nuclear transfer technique for somatic cells that is both simpler and faster than traditional methods. The technique comprises the bisection of zona-free oocytes and the reconstruction of embryos comprising two half cytoplasts and a somatic cell by adherence using phytohaemagglutinin-P (PHA) followed by an electropulse and subsequent culture in microwells (termed WOWs- well of the well). The development of the system was based on results using parthenogenetic and in vitro fertilized zygotes in order to (a) select the optimal primary activation agent that induced the lowest lysis rate but highest parthenogenetic blastocyst yield, (b) evaluate the quantity and quality of zona free blastocysts produced in WOWs, and (c) establish any potential embryotoxic effects of PHA-P. The initial data indicated that, of calcium ionophore A23187, ionomycin, and electropulse treatments as primary activation agents, the two former were equally efficient even with reduced exposure times. WOW-culture of zona-free versus zona-intact zygotes were not different in either blastocyst yield (44.6 +/- 2.4% versus 51.8 +/- 13.5% [mean +/- SEM]) or quality (126.3 +/- 48.4 versus 119.9 +/- 32.6 total cells), and exposure of zygotes to PHA-P did not reduce blastocyst yields compared to vehicle control (40.8 +/- 11.6% versus 47.1 +/- 20.8% of cultured oocytes). Subsequent application of the optimized technique for nuclear transfer using nine different granulosa cell primary cultures (cultured in 0.5% serum for 5-12 days) generated 37.6 +/- 3.9% (11 replicates; range, 16.4-58.1%) blastocysts per successfully fused and surviving reconstructed embryo (after activation), and 33.6 +/- 3.7% blastocysts per attempted reconstructed embryo. Mean day 7 total blastocyst cell numbers from 5 clone families was 128.1 +/- 15.3. The ongoing pregnancy rate of recipients each receiving two nuclear transfer blastocysts is 3/13 (23.1%) recipients pregnant at 5 months after transfer. These results suggest that the zona-free nuclear transfer technique generates blastocysts of equivalent quantity and quality compared to conventional micromanipulation methods, requires less technical expertise, is less time consuming and can double the daily output of reconstructed embryos (even after taking into consideration the rejection of the half oocytes containing the metaphase plate). PMID- 11945224 TI - Sexual maturation and fertility of male Nigerian Dwarf goat (Capra hircus) clones produced by somatic cell nuclear transfer. AB - Three, genetically identical, Nigerian Dwarf bucks produced by somatic cell nuclear transfer (NT) of fetal fibroblasts were monitored for sexual maturation and fertility. Starting at four months of age, these male clones were trained to serve an artificial vagina (AV). Average age of the NT-derived bucks at first semen collection was 20 weeks, which was not different from that of other young bucks of this breed (average age at first collection = 20 weeks). Average sperm production at 5 months of age for the NT-derived bucks was 5.0 x 10(8) spermatozoa, which was comparable to that of dwarf bucks of similar age (3.4 x 10(8) spermatozoa). At seven months of age, semen collected from two NT-derived bucks was used to artificially inseminate six females (three does per buck). Five does were confirmed pregnant by ultrasound at day 42. Nine healthy kids, four males and five females, were born in March and April 2000. Viable spermatozoa were collected from one of the F1 males at 28 weeks of age. These results demonstrated that NT-derived bucks and one of their male offspring developed sexually within the normal timeframe for their breed and that the clones were fertile. PMID- 11945225 TI - Accumulation of the proteolytic marker peptide ubiquitin in the trophoblast of mammalian blastocysts. AB - Ubiquitination is a universal protein degradation pathway in which the molecules of 8.5-kDa proteolytic peptide ubiquitin are covalently attached to the epsilon amino group of the substrate's lysine residues. Little is known about the importance of this highly conserved mechanism for protein recycling in mammalian gametogenesis and fertilization. The data obtained by the students and faculty of the international training course Window to the Zygote 2000 demonstrate the accumulation of ubiquitin-cross-reactive structures in the trophoblast, but not in the inner cell mass of the expanding bovine and mouse blastocysts. This observation suggests that a major burst of ubiquitin-dependent proteolysis occurs in the trophoblast of mammalian peri-implantation embryos. This event may be important for the success of blastocyst hatching, differentiation of embryonic stem cells into soma and germ line, and/or implantation in both naturally conceived and reconstructed mammalian embryos. PMID- 11945226 TI - Human reproductive cloning: premature transfer of a fledgling agricultural biotechnology into human infertility clinics. PMID- 11945228 TI - Application of the zona-free manipulation technique to porcine somatic nuclear transfer. AB - The recent demonstration of a successful zona-free manipulation technique for bovine somatic nuclear transfer (NT) that is both simpler and less labor intensive is of considerable benefit to advance the applications of this technology. Here, we describe that this method is also applicable to porcine somatic NT. Porcine cumulus oocyte complexes were matured in TCM-199 medium before sequential removal of the cumulus and zonae. Zona-free oocytes were bisected using a microknife, and the halves containing the metaphase plate (as determined by Hoechst 33342 staining) were discarded. Each half cytoplast was agglutinated to a single granulosa cell (primary cultures grown in 0.5% serum for 2-5 days prior to use) in phytohaemagglutinin-P. Subsequently, each half cytoplast-granulosa cell couplet was simultaneously electrofused together and to another half cytoplast. Reconstructed embryos were activated in calcium ionophore A23187 followed by DMAP and were then individually cultured in microwells in NCSU 23 medium. On day 7 after activation, blastocyst yield and total cell numbers were counted. Of 279 attempted reconstructed NT embryos, 85.0 +/- 2.8% (mean +/- SEM; n = 5 replicates) successfully fused and survived activation. The blastocyst rate (per successfully fused and surviving embryo) was 4.8 +/- 2.3% (11/236; range, 0-12.8%). Total blastocyst cell count was 36.0 +/- 4.5 (range, 18-58 cells). The blastocyst rate and total cell numbers of parthenogenetically activated and zona-free control oocytes propagated under the same conditions was 11.6 +/- 3.9% (35/335 embryos; n = 3 replicates) and 36.8 +/- 5.2, respectively. Developmentally halted embryos that could still be evaluated on day 7 possessed 54.4 +/- 2.3% (53/96 embryos; n = 3 replicates) anucleate blastomeres, the latter representing 53.5 +/- 6.6% of the blastomeres in such embryos. In conclusion, blastocyst yield was independent of activation efficiency and was likely reduced by insufficient nuclear remodeling, reprogramming, imprinting, or other effects. The data also suggest that fragmentation was a considerable problem that could conceivably contribute to halted development in a high proportion of embryos. The results indicate that the zona-free manipulation technique can be successfully applied to pig somatic NT. Although such zona-free early cleavage stage embryos cannot be transferred to recipients at present, this technique permits simplification of the NT technique for application in basic research, until pig nonsurgical blastocyst transfer becomes a realistic option. PMID- 11945227 TI - Nuclear transfer of blastomeres expressing EGFP-reporter gene may improve the efficiency of transgenic cattle. AB - The effect of timing of microinjection of DNA constructs on the efficiency of transgenic embryo production and improved efficiency and quality through combining EGFP as a reporter gene with nuclear transfer techniques were examined. From 12 to 24 h after insemination, constructs of pCXNeo-EGFP were microinjected into a pronucleus of bovine IVM-IVF zygotes. Due to the difficulty in visualizing pronuclei, the incidence of successful injection of linear DNA was higher when zygotes were injected between 20 and 24 h, as compared with an early period between 12 and 16 h after insemination. However, developmental competence of DNA injected zygotes and the EGFP expression rate were not affected by the injection time. A majority of the embryos expressing EGFP signal were mosaic. Following nuclear transfer of blastomeres expressing EGFP, 4.5% of morulae that developed from the NT eggs had a strong EGFP signal in all live blastomeres. In other embryos, EGFP signal had been lost. When cells derived from the EGFP-positive NT morulae were subcultured, all the cells expressed strong EGFP signal at the second passage and demonstrated neomycin resistance. These results show that transient expression of nonintegrated EGFP appears frequently in EGFP-positive bovine embryos and that additional selection of EGFP-positive morulae after nuclear transfer of EGFP-positive blastomeres would facilitate selection of transgenic embryos. PMID- 11945231 TI - Pronuclear microinjection. AB - Last year marked the 20th anniversary of the invention of the term "transgenic" and the development of pronuclear microinjection, a straightforward technique designed to transfer genetic information from nearly any living organism to mammals. After two decades of use, pronuclear microinjection protocols have changed little from the reliable, if not efficient, method described by Gordon and Ruddle. Experience has taught us that once microinjection skills are perfected there are only a few parameters one needs to be concerned about to successfully produce transgenic animals. Those parameters will be discussed, as will some new innovations that promise to finally increase efficiency of pronuclear microinjection methodology. PMID- 11945230 TI - Nuclear transfer in practice. AB - The technique of nuclear transfer (NT) allows the production of embryos, fetuses, and offspring from a range of embryonic, fetal, and adult derived cell types in a range of species. Successful development is dependent upon numerous factors, including type of recipient cell, source of recipient cell, method of reconstruction, activation, embryo culture, donor cell type, and donor and recipient cell cycle stages. The present review will discuss the uses of NT, the techniques presently available, and the factors affecting subsequent development. PMID- 11945233 TI - The use of recombinase proteins to generate transgenic large animals. AB - The endogenous properties of recombinase proteins allow them to associate with and bind DNA to catalyze homologous recombination. These endogenous properties of cellular recombination enzymes may be useful to the field of transgenesis. The production of transgenic animals, in particular livestock, is an inefficient process by both conventional pronuclear microinjection techniques and nuclear transfer. Furthermore, the use of pronuclear microinjection is currently limited to the random addition of genes and does not allow for the replacement of an endogenous gene with a more desired one. The functions of cellular recombination enzymes have been exploited to develop a technique that is compatible with pronuclear microinjection and may make the process of generating transgenic livestock more efficient while also enabling the targeting of homologous chromosomal genes. In our hands, transgenic animals generated by the pronuclear microinjection of various recombinase protein-coated DNA fragments led to a higher than expected birth rate as well as transgene integration frequency. Most founder animals generated were likely mosaic, indicating that integration occurred after cell division. The presence of multiple related genes makes detection of any recombination event difficult. Overall, this technique is a straightforward, rapid, and efficient procedure that can be applied to any segment of DNA and any microinjection apparatus, and is less labor intensive than nuclear transfer. PMID- 11945232 TI - Gene targeting in primary fetal fibroblasts from sheep and pig. AB - Nuclear transfer offers a new cell-based route for introducing precise genetic modifications in a range of animal species. However, significant challenges, such as establishment of somatic gene targeting techniques, must be overcome before the technology can be applied routinely. In this report, we describe targeted deletion at the GGTA1 (alpha 1,3-galactosyl transferase) and PrP (prion protein) loci in primary fibroblasts from livestock. We place particular emphasis on the growth characteristics of the primary cell cultures, since these are key to determining success. PMID- 11945234 TI - Mammalian artificial chromosomes as vectors: progress and prospects. AB - Artificial chromosomes have long been touted as the ideal vector for gene therapy and biotechnology purposes based on the idea that such a chromosome would mimic the natural state of DNA in the cell. This, it is argued, would mean that essentially unlimited amounts of DNA could be incorporated into such a vector enabling either large genes or whole metabolic pathways to be provided to the recipient cell or organism. Additionally, such a vector would not integrate into the genome of the host cell and so would not cause mutagenesis by insertion and could perhaps be withdrawn from the cell or organism when no longer required. A number of preconditions are implicit in these claims. First, the chromosome should have a segregation efficiency approaching 100% in order to be useful in a cell population undergoing multiple rounds of cell divisions. Second, the chromosome should have a defined structure for regulatory and practical reasons. A defined structure is needed to maximize the control of expression of the genes that it contains. Third, the chromosome should not be so large that delivery becomes a problem. Finally, chromosomal effects such as centromeric or telomeric silencing should not dominate the expression of genes contained in an artificial chromosome. In this article, we discuss our own and others' efforts to achieve these aims using a variety of nonviral approaches to the problem. PMID- 11945235 TI - Cloning, transgenesis, and genetic variance in animals. PMID- 11945237 TI - The action pattern of amylomaltase. PMID- 11945238 TI - Iron-limited growth of torulopsis utilis, and the reversible loss of mitochondrial energy conservation at site 1 and of sensitivity to rotenone and piericidin A. PMID- 11945239 TI - Differences in elastase hydrolysates of elastin from ligamentum nuchae. AB - By means of molecular exclusive chromatography, a marked difference in the distribution of desmosine and isodesmosine in the products of proteolysis has been found between the partial enzymatic hydrolysates of elastin from the bovine nuchal ligament prepared by two different methods. In the preparation which was treated with hot NaOH the prevailing portion was localized in a macromolecular fraction corresponding to the void volume. An increased precipitation of this fraction with trichloracetic acid has been noticed. The autoclaved material forms substantial amounts of the coacervate during the digestion. PMID- 11945240 TI - Effects of thyroidectomy and thyroxine treatment on the activity of 12alpha hydroxylase and of some components of microsomal electron transfer chains in rat liver. PMID- 11945241 TI - Extraction of ribosomal proteins by displacement with protamine. PMID- 11945242 TI - Dinitrophenol and valinomycin as uncouplers in isolated chloroplasts. PMID- 11945243 TI - Stereoselective reactivity of the SH groups of yeast glyceraldehydephosphate dehydrogenase in the allosteric T and R states. AB - Yeast glyceraldehyde-3-phosphate dehydrogenase as a typical SH enzyme is inactivated by the antipodes of a-iodopropionic acid and its amide at different rates. The apoenzyme reacts faster with the D(+) antipode of the free a iodopropionic acid (k(D)/k(L) = 6.8) and the L(-) antipode of the amide (k(L)/k(D) = 3). On addition of NAD(+) the stereoselectivity of the SH group towards a-iodopropionic acid is inverted, that towards the amide is enlarged, the rate relationships depending on the NAD(+) concentration.The results were interpreted by the assumption, that the allosteric T state of the enzyme reacts most rapidly with the D(+) antipodes, whereas the R state favours the L(-) antipodes of the alkylation reagents. The dependence of the reaction rates on the NAD(+) concentration could be fitted to the allosteric function of state R. PMID- 11945244 TI - A spectrophotometric assay for ribonuclease activity using cytidylyl-(3',5') adenosine and uridylyl-(3',5')-adenosine as substrates. PMID- 11945245 TI - Arsenate induced activity of certain enzymes on their dephosphorylated subtrates. PMID- 11945246 TI - Evidence for more than one pathway in the formation of purine deoxyribonucleotides. AB - The biosynthesis of purine deoxyribonucleotides was studied in the context of general purine nucleotide metabolism in the chick with the aid of radioactive nucleic acid precursors. Our results showed that in chick liver and intestine, the nucleoside phosphate reductase system so firmly established in E. coli [1] and L. leichmanni [2] is not exclusively responsible for the biosynthesis of purine deoxyribonucleotides. PMID- 11945247 TI - Origin of mitochondrial enzymes I. Cytochrome c synthesis by endoplasmic reticulum. PMID- 11945249 TI - The isolation of purified brush borders from rat small intestine. PMID- 11945248 TI - Synthesis in vivo of the lecithin component of the inner and outer membranes of rat liver mitochondria. PMID- 11945250 TI - The requirement of the "non-blue" copper (II) for the activity of fungal laccase. PMID- 11945251 TI - The effect of 5-azacytidine on the synthesis of ribosomes in escherichia coli. PMID- 11945252 TI - ? PMID- 11945254 TI - An alkali-soluble glucan fraction from the cell walls of the yeast Saccharomyces carlsbergensis. PMID- 11945253 TI - Activation of L-ornithine by cell-free extracts of Bacillus brevis catalyzing the synthesis of gramicidin S. PMID- 11945255 TI - Proteolytic activity at neutral pH in bovine spleen. AB - Proteolytic activity at neutral pH can be demonstrated in extracts from beef spleen. This activity is completely due to an aminopeptidase - or a number of aminopeptides - which is able to hydrolyse proteins completely to amino acids. No evidence was found for the presence of endopeptidases active at neutral pH. The enzyme resembles to some extent swine kidney aminopeptides. PMID- 11945256 TI - Confirmation of unusual stereochemistry of glutamate biosynthesis in clostridium kluyveri. PMID- 11945257 TI - An alpha(2)-globulin component present in sweat, saliva, tears, human milk, colostrum and cerumen. AB - By double-diffusion and immunoelectrophoresis, using rabbit anti-sweat protein immune serum, an alpha(2)-globulin component of glycoprotein character was found in sweat, saliva, tears, human milk, colostrum and cerumen. PMID- 11945259 TI - Foreword. PMID- 11945258 TI - Isoelectric fractionation of proteins on polyacrylamide gels. PMID- 11945260 TI - Antibodies: Shape, homogeneity, and valency. PMID- 11945261 TI - Wild type variants of penicillinase from Klebsiella aerogenes. PMID- 11945262 TI - Stereospecificity of certain reduced nicotinamide-adenine dinucleotide phosphate linked reactions in rat liver microsomes. PMID- 11945264 TI - Isolation of L-lactyl-puromycin from Bacillus subtilis and yeast. PMID- 11945263 TI - Studies on the bacteriophage MS2. V. The 5'-terminal tetranucleotide sequence of the viral RNA chain. PMID- 11945265 TI - Metal ions and enzyme equilibria: A mathematical treatment. PMID- 11945266 TI - Chain length of ribosomal RNA extracted from mammalian cells. PMID- 11945267 TI - Structure of some peptides isolated from chymotryptic digest of dolphin myoglobin. PMID- 11945268 TI - The disulphide bridges of apamin. PMID- 11945269 TI - A lipid intermediate in the biosynthesis of a teichoic acid. PMID- 11945270 TI - On porcine chymotrypsinogen B. PMID- 11945271 TI - Identification of pantethinase in horse kidney extract. PMID- 11945272 TI - Transient and steady state concentrations of the intermediates of the pentose phosphate pathway in Krebs ascites tumour cells metabolising glucose. PMID- 11945274 TI - Biosynthetic incorporation of cobalt into yeast alcohol dehydrogenase. PMID- 11945273 TI - The reactivities of the lysine, cysteine and tyrosine residues of pig heart lactate dehydrogenase in the presence of sulphite. PMID- 11945275 TI - Some kinetic and molecular properties of yeast phosphofructokinase. AB - Yeast PFK had a sedimentation coefficient of 16.7 S both in the absence and in the presence of ATP, and did not dissociate even at very low protein concentrations. Sodium dodecyl-sulphate caused dissociation of the protein to sub units of 3.2 S.The effects of pH on substrate affinities are described. In the presence of UTP, acting as non-inhibiting phosphate donor, the behaviour of the enzyme towards F-6-P was co-operative, with a Hill coefficient of 2.2. PMID- 11945276 TI - Swelling of rat liver mitochondria with N-(N-acetyl-4-sulphamoylphenyl) maleimide. PMID- 11945277 TI - The molecular weight and subunit structure of glucose-6-phosphate dehydrogenase from human erythrocytes. PMID- 11945278 TI - Amino acid sequence around the reactive cysteine residues in thiolase. PMID- 11945279 TI - Turnover of outer and inner membrane proteins of rat liver mitochondria. PMID- 11945280 TI - ? AB - The influence of the esterification of the 3'(2') hydroxyl group of tRNA on the hydrolysis by venom phosphodiesterase (Crotalus adamanteus) was studied. It was shown that this esterification does not change the rate of hydrolysis. At pH 7-8, no significant differences were observed either when the alpha amino group of the attached amino acid was free or blocked, or when the aminoacid contained a free carboxyl group or a second amino group (glutamic acid or lysine). Aminoacyl-AMP or oligopeptidyl-AMP can be prepared by this method. PMID- 11945281 TI - Partial purification and determination of molecular weights of glyoxalases by filtration on dextran gel. AB - Glyoxalase I and glyoxalase II (EC. 4.4.1.5 and EC 3.1.2.6) were separated by gel filtration on Sephadex G-75 and G-100. This simple procedure permitted also the partial purification of glyoxalase II. The purification coefficient in a single run from supernatant from beef liver was about 1 : 30 compared with 1 : 15 after the fifth step of purification with classical methods. PMID- 11945282 TI - Enzymatic hydrolysis of N-substituted aminoacyl-tRNA. PMID- 11945283 TI - Heterogeneity of yeast mitochondrial fractions revealed by automated analysis of sucrose gradients. PMID- 11945284 TI - ? AB - The nature of the carbohydrate-peptide linkage in the glycopeptides isolated after proteolytic digestion of the structural glycoproteins of the insoluble stroma of porcine and horse aorta was studied. The isolation and identification of 2-acetamido-1-(L-beta-aspartamido)-1,2-dideoxy-beta-D-glucose from an electrophoretically purified glycopeptide proves the presence of aspartamido glucosaminidic linkage in these glycoproteins. PMID- 11945285 TI - Metaphosphate and aerobic CO(2) in Chlorella. PMID- 11945286 TI - Evidence for a three stranded complex between poly I and poly C. AB - The conditions for the formation of a triple stranded semi-protonated complex between polyinosinic (poly (I)) and polycytidylic acid (poly (C)) are described. From potentiometric titrations and mixing curves, the stoichiometry and a phase transition diagram is established between the partly protonated triple stranded and the unprotonated and completely protonated double helical complexes. PMID- 11945287 TI - Metabolism of human erythrocyte glucose-6-phosphate dehydrogenase. III - isolation of a protein involved in the 6-phosphogluconate-induced inactivation. PMID- 11945288 TI - Human erythrocyte AMP: Pyrophosphate phosphoribosyltransferase (EC 2.4.2.7). PMID- 11945289 TI - pH induced changes in optical activity of guanine nucleosides. AB - Optical rotatory dispersion and circular dichroism have been used to investigate the protonation of guanosine and some of its analogues. An inversion of the principal Cotton effect and the dichroic band is observed below the acid pK. It is suggested that a conformational change from the anti form above the pK to the syn form below the pK occurs. The reasons why this change should occur only in guanosine and not in adenosine are discussed. PMID- 11945290 TI - On the site of synthesis of enzymes tightly bound to mitochondrial structure in rat liver. PMID- 11945291 TI - Degradation of encephalitogen by purified brain acid proteinase. PMID- 11945293 TI - A brief survey of some recent work in protein chemistry. PMID- 11945292 TI - Erratum. PMID- 11945294 TI - Mitochondrial biogenesis: Germ-free mitochondria. PMID- 11945295 TI - The effect of ATP on the synthesis of the nicotinamide nucleotides. PMID- 11945296 TI - The effect of heme and iron on haemoglobin A and F synthesis in human cord blood. PMID- 11945297 TI - The sequence around an active-site aspartyl residue in pepsin. PMID- 11945298 TI - Hydrolysis of the glycosidic bonds in beta-O-xylosyl-L-serine and beta-O-D xylosyl-L-threonine by enzyme preparations from Helix pomatia. PMID- 11945299 TI - Isoenzymes of human and rat liver monoamine oxidase. PMID- 11945300 TI - Effect of actinomycin D and puromycin on the conversion of cholesterol into bile acids in bile fistula rats Bile acids and steroids 206. PMID- 11945301 TI - Studies on the stability of protocollagen hydroxylase. PMID- 11945302 TI - Effect of insulin and diet on the steady state concentrations of intermediates of the pentose phosphate pathway of glucose metabolism in liver. PMID- 11945303 TI - Electron microscopic method of ordering the cyanogen bromide peptides of the a 1 chain from acid soluble calfskin collagen. PMID- 11945304 TI - The differences between a 1 and a3 chains of acid soluble calfskin collagen. PMID- 11945306 TI - Interconversion of the components of rat brain mitochondrial malate dehydrogenase. PMID- 11945305 TI - Electrophoretic pattern of and amino acid incorporation in vitro into the insoluble mitochondrial protein of neurospora crassa wild type and mi-1 mutant. PMID- 11945307 TI - Isolation and properties of a phosvitin kinase from chick embryo. PMID- 11945308 TI - Mechanism of action of 2,4-dinitrophenol on the soluble atpase from mitochondria. PMID- 11945310 TI - Uptake of 2,4-dinitrophenol by anaerobic yeast cells and its relation to the energy transduction in these cells. PMID- 11945309 TI - Synthesis of labelled ATP from ADP and labelled pyrophosphate by spinach chloroplasts. PMID- 11945311 TI - The control of mitochondrial enzyme synthesis in yeast: A new hypothesis. PMID- 11945312 TI - Separation of two initiator transfer RNAs from E. coli. PMID- 11945313 TI - Isolation and properties of group specific 2-oxoglutarate aminotransferase from Streptomyces noursei 2/9. PMID- 11945315 TI - Erratum. PMID- 11945314 TI - Exchange reaction between ATP and labelled pyrophosphate by isolated spinach chloroplasts. PMID- 11945316 TI - 14alpha-Hydroxylation of testosterone and androst-4-ene-3,17-dione in rat liver. PMID- 11945317 TI - The inability of thiols to reduce cobalamins in the absence of a metalion. PMID- 11945318 TI - 'Paradox' effect of cortisol and actinomycin D on RNA polymerase activity of rat liver nuclei. PMID- 11945319 TI - The fractionation of ribonucleic acid on a preparative scale by polyacrylamide gel electrophoresis. PMID- 11945320 TI - Enzymic conversion of linolenic acid to ethylene by extracts of apple fruits. PMID- 11945321 TI - Glycine as crosslinking bridge in the LL-diaminopimelic acid containing murein of Propionibacterium peterssonii. PMID- 11945322 TI - ? AB - A method for the preparation of UDP-glucose from HClO(4)-extract of diaphragm using paper-chromatography is described. It allows the quantitative determination of UDP-Glc by optical test and the measurement of its specific (14-C) activity simultaneously. PMID- 11945323 TI - Kinetic evidence for an enzyme-bound intermediate in the biosynthesis of a methionine. PMID- 11945324 TI - Influence of spermine and histones on a nuclear nucleosidase with ADP-ribose transferase activity in ehrlich ascites cells. PMID- 11945325 TI - Conformational-transitions of bovine pancreatic ribonuclease S-peptide. PMID- 11945326 TI - Regulation of NADH and NADPH metabolism in Azotobacter vinelandhii. PMID- 11945327 TI - ? AB - A frog (Rana esculenta) hemoglobin has been purified and two types of chain have been separated by countercurrent distribution, N- and C-terminal sequences of each chain have been determines. The N-terminal amino acids are Acetyl-Ala and Gly. The acetylated chain seems to belong to the alpha-chain series and the glycine chain to the beta-chain series. PMID- 11945328 TI - The ferritin content of the free ribosome fraction isolated from adult male and female rat liver. PMID- 11945329 TI - Degradation studies on the lipopolysaccharide from E. coli 071:K?:H12. Separation and investigation of O-specific and core polysaccharides. PMID- 11945330 TI - Iodine content of submitochondrial particles prepared from rat liver by drastics sonication. PMID- 11945331 TI - Allosteric glutaminase in rat liver of likely regulatory significance. PMID- 11945332 TI - "Solubilization" of the stearoyl-CoA desaturase of rat liver microsomes. PMID- 11945333 TI - Studies on alpha-naphthol as a precursor of microbial menaquinone. PMID- 11945334 TI - The synthesis of nucleoside 2'(3')-phosphate 5'-triphosphates. PMID- 11945335 TI - Osmotic pressure studies on the association of acid-extracted "histone IIb" PMID- 11945336 TI - Binding of salicylate to albumin. PMID- 11945337 TI - The kinetics of reaction of bovine thrombin with p-nitrophenyl p' guanidinobenzoate and 7-amino-1-chloro-3-toluene-p-sulphonamido-2-butanone: A new method for determining the operational molarity of thrombin solutions. PMID- 11945338 TI - A kinetic interpretation of the allosteric model of Monod, Wyman, and Changeux. PMID- 11945339 TI - Photoreduction of flavocoenzymes by pyruvic acid. PMID- 11945341 TI - Antagonistic homotropic interactions as a possible explanation of coenzyme activation of glutamate dehydrogenase. PMID- 11945340 TI - The presence of protein bound intermediates in the biosynthesis of gramicidin S. PMID- 11945342 TI - Errata. PMID- 11945344 TI - The applicability of the donnan relation to the distribution of certain anions between mitochondria and medium. PMID- 11945343 TI - Calcium transport in mitochondria. PMID- 11945345 TI - Nucleophilic sites in energized mitochondrial membranes. PMID- 11945346 TI - ? AB - Leucine aminopeptidase of bovine lens is a zinc metallo-enzyme. A zinc content of 5.0 - 7.6 g-atoms of Zn/M.W. 3.2 x 10(6) was estimated by labelling with 65Zn and by atomic absorption spectrophotometry. By continuous dialysis against o phenanthroline zinc was almost completely removed. The enzymatically inactive "apo"-enzyme could not be reactivated by addition of Zn(2+). The binding of cadmium, manganeous and cobaltous ions was investigated. With some probability manganese is bound at an other site than zinc. The effect of pH and buffer ions on the binding of zinc to leucine aminopeptidase was demonstrated. PMID- 11945347 TI - Hydrogen peroxide as the product of autoxidation of ferredoxin: Reduced either chemically or by illuminated chloroplasts. PMID- 11945348 TI - Mitochondrial peptide chain elongation factors from Neurospora crassa. AB - Two complementary peptide chain elongation factors (G and T) have been isolated from a mitochondrial 100,000 g supernatant. Both factors are specific for 70 S ribosomes and can be crossed with T and G factors from E. coli. PMID- 11945349 TI - Structural specificity for the photoinactivation of nucleic acids by furocoumarins. PMID- 11945350 TI - Role of bivalent cations in the control of enzymes involved in gluconeogenesis. PMID- 11945351 TI - Peptidyl-tRNA and peptidyl transferase activity of skeletal muscle ribosomes. Effect of diabetes. PMID- 11945352 TI - Structural and spectral properties of 2,4-dithiouridine. PMID- 11945353 TI - On the isolation, characterisation, and crystallisation of a human Bence-Jones protein of kappa type. AB - The isolation and purification of the human Bence-Jones protein Rei is described. Physico-chemical data are given characterising the protein. It is shown that the protein could be crystallised. It is intended to use the crystals for single crystal X-ray diffraction studies. PMID- 11945354 TI - Amino acid sequence homology in the active site of rabbit and sturgeon muscle aldolases. PMID- 11945355 TI - The effect of rifampicin on mitochondrial RNA polymerase from rat liver. PMID- 11945356 TI - ? AB - A method of isolation of the isoenzyme BB of creatine kinase from ox stomach is described taking into account its high instability and the presence of notable amounts of tropomyosin in vertebrate smooth muscle extracts of low ionic strength. The molecular parameters of the enzyme isolated from ox stomach do not differ from those of the creatine kinases MM and BB of vertebrates. The amino acid composition and the peptide maps reveals significant differences from that of the ox brain isoenzyme BB. PMID- 11945357 TI - A glutamine-dependent asparagine synthetase from yellow lupine seedlings. PMID- 11945359 TI - Erratum. PMID- 11945358 TI - Some regulatory properties of cytidine deaminase and uridine phosphorylase of Bacillus cereus. PMID- 11945360 TI - The molecular basis of recombination a report of the meeting of the British biophysical society held at the University of Sussex, March 24-25, 1970. PMID- 11945361 TI - Diurnal incorporation of 3H-leucine into liver protein. PMID- 11945362 TI - ? AB - 5'-Methylammonium-5'-deoxyadenosine is a poor subsrate for adenosine desaminase. The nucleoside was crystallized from aqueous methanol as its iodide monohydrate in space group P2(1). The structure was solved from three dimensional X-ray data and refined to R = 4.1%. The structure of the nucleoside is determined by an intramolecular hydrogen bond from N5' to N3 which implies that (1) the edenine heterocycle is in syn position with respect to the sugar moiety; (2) the ribose pucker is C2'-exo, C3'-endo and (3) the conformation about the C4'-C5' bond is trans, gauche. The water of hydration is fourfold disordered and forms a linear hydrophilic region which is surrounded by nucleoside molecules in a zigzag arrangement. The adenine heterocycles are not stacked and in closer than Van der Waals distance with iodine ions suggesting a charge transfer type interaction. PMID- 11945363 TI - Effects of L-asparaginase on protein and glycoprotein synthesis. PMID- 11945364 TI - Peptide chain initiation in a species of Nostoc and in chloroplasts of Euglena gracilis. PMID- 11945365 TI - Energy-linked reactions in mitochondria: A requirement for ubiquinone after pentane extraction. PMID- 11945366 TI - Activation of mitochondrial DPN kinase from yeast. PMID- 11945367 TI - Interaction of rat muscle AMP aminohydrolase with chelating agents and metal ions. PMID- 11945368 TI - Phosphorylation of some rat liver ribosomal proteins and its activation by cyclic AMP. PMID- 11945369 TI - Evidence for the existence of an alpha-glycerophosphate oxidase system with three phosphorylation sites and sensitive to rotenone and piericidin A. PMID- 11945370 TI - The role of rat liver nuclear DNA polymerase and its distribution in various classes of liver nuclei. PMID- 11945371 TI - Long-chain triglyceride lipases of pig liver. PMID- 11945372 TI - Elongation factors from human lymphatic tissue: Isolation and some properties. PMID- 11945374 TI - Acetone-sensitized anaerobic photo-oxidation of methionine. PMID- 11945375 TI - Errata. PMID- 11945373 TI - Formation of persulfide groups in alkaline treated insulin. PMID- 11945376 TI - ? AB - Disintegration or altering of intact mitochondria or digitonin inner membrane + matrix particles causes an increase in K(+)-linked release of protons. This effect is not obtainable in the case of outer membranes. It is assumed, that also the inner membrane + matrix possesses exchange sites for K(+), which are freely accessible for the potassium ion if the structural integrity is damaged. The outer layer of inner membrane seems to be a barrier for the K(+)-movement. PMID- 11945377 TI - The beta-chain of frog hemoglobin (Rana esculenta): The complete amino acid sequence. PMID- 11945378 TI - Effect of some organometallic compounds on the permeability of chloroplast membranes. PMID- 11945379 TI - Specific transport of inorganic phosphate, 3-phosphoglycerate and dihydroxyacetonephosphate, and of dicarboxylates across the inner membrane of spinach chloroplasts. PMID- 11945380 TI - Chloroplast and cytoplasmic ribosomes in Euglena gracilis. PMID- 11945381 TI - Occurrence of termination ribosomes as free 70 S particles. PMID- 11945382 TI - Transient repression of the lac operon - the effect of a lac promoter deletion. AB - Experiments have been done to show whether the lac promoter delection L1, which partly alleviates catabolite repression, also affects transient repression of lac. In stain L1/F'M15 all of the beta-galactosidase is synthesized from a chromosomal gene cis to L1, whereas 98% of the thiogalactosidase transacetylase is synthesized from an episomal gene cis to an intact i-p-o region. The addition of glucose to induced cultures of strain L1/F'M15 growing in glycerol medium caused extensive transient repression of transacetylase but almost no transient repression of beta-galactosidase. In control experiments with a diploid stain of genotype p(+)z(+)a(-)/F'p(+)z(-)a(+) the two enzymes suffered equal transient repression. Thus L1 substantially relieves transient repression. PMID- 11945383 TI - Separation of two different peptidyl transfer RNA translocases from mammalian tissues. PMID- 11945384 TI - Specificity of factors required for peptide elongation in mammalian cells. PMID- 11945385 TI - RNA synthesis and the stimulation of insulin biosynthesis by glucose. PMID- 11945386 TI - Cd(2+), Mn(2+) and Zn(2+) induced synthesis of nuclear RNA in the livers of normal and adrenalectomized rats. PMID- 11945387 TI - Studies on the complex of distamycin a with calf thymus DNA. PMID- 11945388 TI - The redox potential of cytochrome c: Ion binding and oxidation state as linked functions. PMID- 11945389 TI - Reactivating effect of alpha,omega-bis-(4-pyridinealdoxime)-2-trans-butene dibromide on isopropyl-methylphosphonylated acetylcholinesterase. PMID- 11945390 TI - Cycle dependent variation of X-ray effects on synchronous mitosis and thymidine kinase induction in Physarum polycephalum. PMID- 11945391 TI - Pullulan, a relationship between molecular weight and fine structure. PMID- 11945392 TI - Subfragment 1 of cow carotid myosin. PMID- 11945393 TI - The histopathological effects of aflatoxin B(1) and the palmotoxins B(0) and G(0) on the liver of the developing chick embryo. PMID- 11945394 TI - Oligomycin resistant mutants in yeast. PMID- 11945395 TI - The effect of glucagon on the state of lysosomal enzymes in isolated perfused rat liver. PMID- 11945396 TI - The effect of a purified preparation of lecithinase C on iodide transport. AB - Treatment of porcine thyroid slices with highly purified lecithinase C leads to a marked reduction in the ability to accumulate iodide ion. Although this response is less sensitive as a function of lecithinase concentration than is the ability to respond to thyrotropin with an increase in glucose oxidation, phospholipid synthesis or adenyl cyclase activity, it occurs when the baseline values of these parameters are not substantially altered. PMID- 11945397 TI - Specificity of pepsin: Size and property of the active site. PMID- 11945398 TI - Behaviour of phenylalanyl-tRMA from Halobacterium cutirubrum on BD-cellulose. PMID- 11945399 TI - Exposed cytosine residues in the tRNA(Val)(1) from yeast. PMID- 11945400 TI - On the reversibility of the biochemical reactions of muscular contraction during the absorption of negative work. PMID- 11945401 TI - Lack of concordance between primary structure and antigenicity in the case of various lysozymes and of bovine alpha-lactalbumin. PMID- 11945402 TI - Fluorescence study of the binding dynamics of saccharides to lysozymes. PMID- 11945403 TI - Respiratory control in membrane particles from Micrococcus denitrificans. PMID- 11945404 TI - Isolation of 5,6-dimethylbenzimidazolyl cobamide coenzyme from Rhodopseudomonas spheroides. PMID- 11945405 TI - Increased rate of cyclic photophophorylation in preparations from Anabaena variabilis cells grown in the presence of diphenylamine. AB - Cell free homogenates and membrane fractions prepared from Anabaena variabilis cells grown in the presence of diphenylamine have markedly higher activities for cyclic phosphorylation than similar preparations from normal cells. The preparations from diphenylamine-grown cells are also more active in system I mediated electron transport from reduced dichloroindophenol to oxygen or methyl viologen. The light intensity required to saturate phenazine methosulphate supported cyclic phosphorylation, in such preparations, is higher than for preparations for normal cells. PMID- 11945406 TI - Solubilized DNA-dependent RNA polymerase from human placenta: A Mn(2+)-dependent enzyme. AB - The 100-fold purified RNA polymerase activity from human placenta is completely dependent upon added DNA. The enzyme is most active at 3 mM Mn(2+) in the presence of 100 mM (NH(4))(2)SO(4). Denatured DNA is a better template than native DNA. alpha-Amanitin completely inhibits the incorporation of 3H-UMP, while rifampicin has no influence upon the enzymatic activity. PMID- 11945407 TI - pH-dependent changes in density of plasma membranes of growing Mycoplasma laidlawii cells. PMID- 11945408 TI - Differential inhibition of phosphate efflux and influx and a possible discrimination between an inner and outer location of the phosphate carrier in mitochondria. PMID- 11945409 TI - Stimulation of mitochondrial RNA synthesis by thyroid hormone. PMID- 11945410 TI - Protein syntheis in brain mitochondrial and synaptosomal preparations. AB - The anomolous protein synthesis, sensitive to cycloheximide, which has been observed in brain mitochondrial and synaptosomal preparations, has been studied. It is concluded that this protein synthesis is due to the presence, as a contaminant in both preparations, of a ribosome-containing particle which contains soluble enzymes and is limited by a plasma membrane. PMID- 11945411 TI - The proteins of the inner membrane of rat liver mitchondria. AB - Abteilung MolekulareBiologie, Max-Planck-Institut Experimentelle Medizin, Gottingen, Germany PMID- 11945412 TI - The biosynthesis of a blood group active tetrasaccharide. PMID- 11945413 TI - Role of initiation factor B. PMID- 11945415 TI - The formation of acetate from pyruvate in house fly flight muscle mitochondria. PMID- 11945414 TI - The physicochemical basis of ion transport through biological membranes. Report of a Pre-symposium held in Riga, U.S.S.R. on June 19 and 20, 1970, within the scope of the VII. International Symposium on the Chemistry of Natural Products. PMID- 11945416 TI - On the proton translocation system of the inner mitochondrial membrane. PMID- 11945417 TI - Proton uptake and phosphorylation in digitonin-treated chloroplast particles. PMID- 11945418 TI - The effects of synthetic glucocorticosteroid (triamcinolone diacetate) and dibutyryladenosine-3',5'-(cyclic) monophosphate on hepatic lipase activities in rats. PMID- 11945420 TI - Uptake of salicylic acid into mycobactin S by growing cells of Mycobacterium smegmatis. PMID- 11945419 TI - Cytoplasmic location of certain small molecules after their metabolism by the endoplasmic reticulum of rat liver. PMID- 11945421 TI - Soybean trypsin inhibitor-C: An active derivative of soybean trypsin inhibitor composed of two noncovalently bonded peptide fragments. PMID- 11945422 TI - The fatty acids of myelin phospholipids. PMID- 11945423 TI - Common and specific determinants on the Fd fragments of guinea-pig immunoglobulins. PMID- 11945424 TI - Preparation and properties of urease chemically attached to nylon tube. PMID- 11945425 TI - Phosphorylase kinase deficiency in mice. PMID- 11945426 TI - Purification, characterization and crystallization of pork myokinase. PMID- 11945427 TI - Optical rotatory dispersion properties of nucleic acid complexes with the oligopeptide antibiotics distamycin A and netropsin. AB - ORD measurements of nucleic acids in the presence of the oligopeptides netropsin and distamycin A have indicated association of the antibiotics with DNA and strong conformational changes of the DNA structure with specificity to AT-rich helical regions. The RNA conformation is relatively unaffected by these antibiotics. The results are explained in terms of a perturbation of the DNA secondary structure as well as of the chromophore system of the oligopeptides. PMID- 11945428 TI - Oxidation of a variety of natural electron donors by the thiol-oxidising agent, diamide. PMID- 11945429 TI - The quantitative dependence of glucose-6-phosphohydrolase upon phospholipids: Effects of phospholipase C at 50 degrees and 370 degrees. PMID- 11945430 TI - Scorpion neurotoxins: A family of homologous proteins. PMID- 11945432 TI - Addendum. PMID- 11945433 TI - Errata. PMID- 11945431 TI - Resolution of five conformational transitions in phenylalaninespecific tRNA from yeast. PMID- 11945434 TI - Antibiotics: Chemistry and mode of action Report of a Pre-Symposium held in Riga, U.S.S.R., on June 19 and 20, 1970, within the scope of the VII. International Symposium on the Chemistry of Natural Products. PMID- 11945435 TI - Effects of glucose and insulin on lipolysis rates in human fat cells of different sizes. PMID- 11945436 TI - Synthesis and properties of poly-2,4-dithiouridylic acid, a new analog of poly uridylic acid. PMID- 11945437 TI - The quantitative determination of tyrosine, 3-iodotyrosine and 3,5-diiodotyrosine in iodinated insulin preparations. PMID- 11945438 TI - Esters of iodinated tyrosine as inhibitors of chymotrypsin. PMID- 11945439 TI - An in vitro effect of thyroid hormone upon bone marrow synthesis of hemoglobin. PMID- 11945440 TI - Some properties of a fatty acid omega-hydroxylation system solubilized from liver microsomes. PMID- 11945441 TI - Analogy between in vivo and in vitro biological effect of chloramphenicol and its acetylated derivatives. PMID- 11945443 TI - Envelope protein changes in mutants of Escherichia coli refractory to colicin E2. PMID- 11945442 TI - Glycopeptides from plasma membranes and microsomes of rat liver. PMID- 11945444 TI - Isolation of a "basic membrane" fraction enriched in an ornithine-containing lipid, from a blue-green mutant of Rhodospirillum rubrum. PMID- 11945445 TI - Methane oxidation by cell-free extracts of Methylococcus capsulatus. PMID- 11945446 TI - Epigenetic interconversions of the multiple forms of mouse liver catalase. PMID- 11945447 TI - Fluorescence studies on the 30 S ribosome assembly process. PMID- 11945448 TI - Phosphorolysis of tRNA. Conformation of yeast tRNA(Phe)(HCl) and the recombined molecules with 3' and 5' halves. PMID- 11945449 TI - Yeast mitochondrial ribosomal RNA: A new extraction procedure and unusual physical properties. PMID- 11945450 TI - Formation of coenzyme a esters of cinnamic acids with an enzyme preparation from cell suspension cultures of parsley. AB - The formation of coenzyme A thiol esters of cinnamic, p-coumaric, p-methoxy cinnamic, and ferulic acids was catalyzed by enzyme preparations from cell suspension cultures of leaf petioles from parsley (Petroselinum hortense Hoffm.). Of these acids, p-coumaric acid served as the most efficient substrate. Enzyme activity is markedly increased upon illumination with white light in a manner very similar to that in which the activities of a number of enzymes involved in flavone biosynthesis are stimulated by light. This strongly suggests that the formation of p-coumaroyl coenzyme A is part of this biosynthetic pathway. PMID- 11945451 TI - In situ oxidation of lactate by skeletal muscle during intermittent exercise. PMID- 11945452 TI - Phosphorylation of skeletal muscle microsomes by acetylphosphate. AB - The phosporyl group of acetylphosphate is transferred to the membranal protein of the sarcoplasmic vesicles during active calcium transport. Although the phosphoprotein formed cannot be distinguished from that obtained in the presence of ATP, the conditions for ATP and acetylphosphate hydrolysis are different from each other. PMID- 11945453 TI - tRNA: Structure and functions Report of a Pre-Symposium held in Riga, U.S.S.R., on June 19 and 20, 1970, within the scope of the VII International Symposium on the Chemistry of Natural Products. AB - The four sessions of the Pre-symposium were devoted to four different aspects of the subject, these form the main divisions of the report. The Pre-symposium was opened with an introductory speech by V.A.Engelhardt. PMID- 11945454 TI - Studies on the half life time of rat liver transfer RNA species. AB - The tRNA fraction, extracted from very high speed supernatant fluid, from livers of rats injected with 3H-orotic acid, attained maximum specific activity after a little over 24 hr and, thereafter, decayed with an apparent half life of 5 days. This behaviour of tRNA was indistinguishable from that of liver rRNA and the acid soluble pool. Chromatography of tRNA, doubly labelled during a period of short synthesis and of prolonged decay, on a BD cellulose column, indicated that individual tRNA species turn over at a constant rate with respect to one another. PMID- 11945455 TI - A simplified method for purification of yeast "renaturable" leucine transfer RNA: The nucleotide sequence of fragments obtained by pancreatic ribonuclease digestion. PMID- 11945456 TI - A comparison of the primary structures of the 16 S ribosomal RNAs from Escherichia coli and Proteus vulgaris. PMID- 11945457 TI - On the structure of a peptido-phosphogalactomannan complex from a black yeast, Cladosporium werneckii. PMID- 11945459 TI - Isoelectric focusing of fibrinogens on polyacrylamide gels. PMID- 11945458 TI - Biosynthesis of yeast mannan: Inhibition of synthesis of mannose acceptor by cycloheximide. PMID- 11945460 TI - Evidence for a new coagulation factor. PMID- 11945461 TI - Direct evidence for a correlation between amino acid sequence and cross striation pattern of collagen. PMID- 11945462 TI - ? AB - The A variant of bovine alpha(s1) casein is devoid of the segment of 13 amino acid residues which occupies the 14th to 26th position from the NH(2)-terminal in the polypeptide chain (198 residues) of the B and C variants. PMID- 11945463 TI - Minor constituents of human milk II. Identification of dodecadienoic-, tetradecadienoic- and hexadecadienoic acid in human milk fat. PMID- 11945464 TI - Alterations in balance of long chain metabolic derivatives of linolenic acid in the fat deficient guinea pig. PMID- 11945465 TI - ? AB - Isolated plastids (chloroplasts and etioplasts) that are morphogically intact and free of any contamination from other cell structures can incorporate label from sn-glycerol-3-phosphate-(14)C into monogalactosyldiglyceride. PMID- 11945466 TI - On the interaction of ATP with the phosphorylation system in chloroplasts. PMID- 11945467 TI - Steroids in primates: Excretion of 3beta-hydroxy-Delta(5) steroids by a newborn chimpanzee (Pan Troglodytes). PMID- 11945468 TI - The hepatotoxic action of carbon tetrachloride stimulatory effect of carbon tetrachloride on lipid peroxidation in microsomal suspensions. PMID- 11945469 TI - The composition of the rabbit muscle triosephosphate isomerase isozymes of two different subunits. PMID- 11945470 TI - Inactivation of metalloenzymes by salicylate. PMID- 11945472 TI - Erratum. PMID- 11945471 TI - L-aspartate aminotransferase: Protection against the formation of multiple forms. PMID- 11945473 TI - Inhibition by thiostrepton of the formation of a ribosome-bound guanine nucleotide complex. PMID- 11945474 TI - Studies on translocation V: Fusidic acid stabilization of a eukaryotic ribosome translocation factor-GDP complex. PMID- 11945475 TI - Association of subunits in purified preparations of Escherichia coli ribosomes. PMID- 11945476 TI - A specific chemical chain scission of tRNA at 7-methylguanosine. PMID- 11945477 TI - Reconstitution of bacterial DNA-dependent RNA-polymerase from isolated subunits as a tool for the elucidation of the role of the subunits in transcription. PMID- 11945478 TI - On the liberation of sigma and the molecular weight of E. coli RNA polymerase. PMID- 11945479 TI - The steady state level of catalase compound I in isolated hemoglobin-free perfused rat liver. PMID- 11945480 TI - ? AB - The contents of sterol derivatives (sterols + esters, glycosides and acylated glycosides) have been determined using thin layer chromatography in leaves and bulky and organs from carrots (Daucus carota L.) potato (Solanum tuberosum L.), onion (Allium cepa L.) two varieties of radish (Raphanus sativus L.) and Gentiana purpurea L. plants. Ether-soluble lipids of leaves contain 2.8-5.2%, those of storage tissue 3.1-8.9% sterol derivatives. The fractionation of lettuce (Lactuca sativa L., Burpee's Ruby Red) leaf homogenates indicates steryl glycosides to be accumulated in a microsomal fraction. PMID- 11945481 TI - Ganglioside patterns of three morris minimal deviation hepatomas. PMID- 11945482 TI - Cyclic 3',5'-adenosine monophosphate-mediated follicular reorganization of isolated thyroid cells in culture. PMID- 11945483 TI - Crystallization of orcinol hydroxylase from Pseudomonas putida. PMID- 11945484 TI - Trapping of uridine phosphates by D-galactose in ethanol-treated liver. PMID- 11945485 TI - Heterogeneity of mitochondria in rat brain. PMID- 11945486 TI - The effect of p-hydroxymercuribenzoate on mitochondrial anion exchange reactions involving L-malate. PMID- 11945487 TI - Energy transfer to cytochrome c in an artificial lamellar system. PMID- 11945488 TI - Regeneration of native structure and biological activity by air oxidation of reduced pancreatic secretory trypsin inhibitor and Des-Thr-Ser-Pro-Gln-Arg inhibitor. PMID- 11945489 TI - Intramolecular reaction of specific substrate as a step in alpha-chymotrypsin catalyzed hydrolysis. PMID- 11945490 TI - Intramolecular dehydratation of phenylthiocarbamyl aminoacids induced by alpha chymotrypsin. PMID- 11945492 TI - The relation between mitochondrial anion accumulation and the pH gradient with special reference to phosphate distribution. PMID- 11945491 TI - Proflavine inhibition of protein synthesis in malignant hamster melanoma. PMID- 11945493 TI - The effect of some anions on respiration inhibited by progesterone. PMID- 11945494 TI - Degradation of mitochondrial respiratory system of Saccharomyces cerevisiae, induced by respiratory inhibitors, uncouplers and anaerobiosis. PMID- 11945496 TI - Stacked and extended conformations of single stranded polynucleotide chain favoured by van der Waals forces. PMID- 11945495 TI - A model of enzymatic synthesis of the internucleotide bond between oligodeoxynucleotides. PMID- 11945497 TI - ? AB - Several authors have described the NBD-chloride (7-chloro-4-nitrobenzo-2-oxa-1,3 diazole) as a fluorogenic reagent of the amino groups and the sulfhydryl groups. It reacts rapidly with the trypsin to produce a stable and highly fluorescent conjugate. The determination of the labelled residues was made with thin layer chromatographies; the lysin residues were preferentially labelled.Fluorescence depolarization measurements of the conjugate were used to determine the brownian relaxation of the enzyme. The temperature dependence of the life-time is very strong and the corrected Perrin's plot demonstrates that the relaxation time of the enzyme decreases rapidly with the temperature.The conclusion is that the NBD chloride can be used, like the DNS-chloride but is more reactive and stable and its fixation, on trypsin can be followed by measuring the absorbance of the dye at 470 nm. PMID- 11945498 TI - ? AB - Tryptic hydrolysis of different arginine nitroanilides was studied. Para nitroanilide is the most readily hydrolyzed substrate, ortho derivative is 5 times less sensitive and meta derivative is a very weak substrate. When the benzoyl group bound on the N(alpha) arginine of BAPNA is replaced by a carbobenzoxy group (L-ZAPNA), the tryptic hydrolysis is increased by a factor of 3. With this latter substrate, the kinetic constants are determined for two forms, native and Nepsilon-acetylated of porcine and bovine trypsin. Evidence is presented that the rate of hydrolysis is both related to the global and local charges of the enzyme. PMID- 11945500 TI - Physical characterization of the DNA released from phage particles by heat inactivation. PMID- 11945499 TI - Sea urchin satellite DNA. Its isolation, some properties and relative amount at different stages of sea urchin development. PMID- 11945501 TI - Proteins and glycoproteins of rat cerebral cortex subsynaptosomal fractions: Extraction with sodium dodecylsulphate and analytic electrophoresis. PMID- 11945502 TI - The relative configuration of N(5)-methyl-L-tetrahydrofolic acid. PMID- 11945503 TI - Metabolism of 3-deoxy-3-fluoro-D-glucose by Pseudomonas fluorescens. PMID- 11945504 TI - The formation and catabolism of methylglyoxal during glycolysis in Escherichia coli. PMID- 11945505 TI - Kinetic response of a membrane-bound acetylcholinesterase to cholinergic activating and blocking agents. PMID- 11945507 TI - Allosteric inhibition of muscle pyruvate kinase by phenylalanine. PMID- 11945506 TI - On the active site of elastase: Partial mapping by means of specific peptide substrates. AB - RNase-S peptide as well as some related octa- and hexapeptides were found to be highly, reactive substrates of porcine elastase (e.g. Ala(4)-Lys-Phe: K(m) = 4500 M(-1), k(cat) = 32 sec(-1), C = 1.4 x 10(5) M(-1) sec(-1)). Comparison of the various peptides led to the conclusion that the active site of porcine elastase is composed of 6-7 subsites (c.f. [1]). Preliminary mapping shows that subsites S(2), S'(1) and S'(2) have hydrophobic character. Occupation of subsite S(4) by the substrate is important for efficient hydrolysis. Binding at this subsite was found to be stereospecific. PMID- 11945508 TI - Analogy between active sites of cathepsin B(1) and papain. PMID- 11945509 TI - Cytidylic and adenylic acid incorporation into fragments of tRNA. PMID- 11945510 TI - A fluorescent probe for the dimer to tetramer conversion of glycogen phosphorylases a and b. PMID- 11945512 TI - Electric fields in coupling membranes. PMID- 11945511 TI - Determination of C-terminal amino acid residues of bovine fibrinogen by hydrazinolysis. PMID- 11945513 TI - Some properties of dinitrofluorobenzene-treated 50 S ribosomal subunits. PMID- 11945514 TI - Formation of initiation complex with lambdamRNA in vitro. AB - The product from the in vitro transcription of native lambdaDNA by purified E. coli RNA polymerase does not stimulate total amino acid incorporation although it can program ribosomes for the first peptide bond synthesis in the presence of the three initiation factor A, B and C. PMID- 11945515 TI - TYMV RNA As a substrate of the tRNA nucleotidyltransferase. PMID- 11945516 TI - Interactions of seryl-tRNA synthetase with serine and phenylalanine specific tRNA. PMID- 11945517 TI - Ketogenic action of fructose in the isolated perfused rat and guinea pig liver. PMID- 11945518 TI - Citrate degradation by ascorbate and cupric or ferrous ion. PMID- 11945519 TI - Molecular evolution of the neurohypophysial hormones: The active peptides of a primitive bony fish Polypterus bichir. AB - Neurohypophysial hormones have been so far identified in Neopterygii and Crossopterygii but not in species of the bird sub-class of bony fishes, the Palaeopterygii. Isolation and chemical characterization of the active principles of a primitive bony fish, Polypterus bichir, have been performed. Isotocin (Ser(4)-Ile(8)-oxytocin) and arginine vasotocin (Arg(8)-oxytocin) have been identified. Because the same peptides were found in the recent Neopterygii, it can be deduced that neurohypophysial hormones have displayed a peculiar stability in the course of the evolution of bony fishes. However isotocin and vasotocin are replaced by oxytocin and vasopressins in mammals and therefore might be regarded as "old" molecules. PMID- 11945520 TI - Purification of Bacillus pumilus beta-D-xylosidase by affinity chromatography. PMID- 11945521 TI - Expression of allelic and non-allelic genes in immunoglobulin light chains from homozygous rabbits. PMID- 11945522 TI - The hydrolysis of Tay-Sachs ganglioside (TSG) by human N-acetyl-beta-D hexosaminidase A. PMID- 11945524 TI - On the mechanism of gramicidin S formation from intermediate peptides. PMID- 11945525 TI - Errata. PMID- 11945526 TI - Binding of the in vitro synthesized RNA to rat liver 40 S ribosomal subunit. PMID- 11945527 TI - Dissociation of 70 S E. coli ribosomes induced by a ribosomal factor (DF). Electrophoretic studies of the ribosomal particles. AB - The dissociation of purified 70 S.E. coli ribosomes, induced by the dissociation factor DF, has been studied by submitting the reaction mixtures to electrophoresis on polyacrylamide gels. The electrophoretic analysis of the ribosome mixtures revealed a heterogeneity which escaped detection by conventional sucrose gradient centrifugation. Increasing amounts of DF in the reaction mixtures converted 70 S ribosomes to particles (designated 70 S (I)) which migrate slower in the electric field than the original 70 S ribosomes. These 70 S (I) ribosomes still consist of both subunits. They dissociate upon further raising the DF concentration. PMID- 11945528 TI - Effects of tris and oligoamines on the ribosomal size distribution in lysates of B. licheniformis. PMID- 11945529 TI - Induction of autohemolysin formation in spleen cell culture by "immunogenic RNA" PMID- 11945530 TI - Effects of dihydrostreptomycin treatment in vivo on the ribosome cycle in Escherichia coli. PMID- 11945532 TI - Preparation of a pG-fragment from tRNA(Phe)(yeast) by chemical scission at the dihydrouracil, and inhibition of tRNA(Phe)(yeast) charging by this fragment when combined with the -CCA half of this tRNA. PMID- 11945531 TI - Localization of messenger RNA near the 3'-end of the dRNA precursor molecule. PMID- 11945533 TI - Competitive inhibition of the acceptor activity of tRNA(Tyr II)(E. coli) by a combination of oligo G and a CCA terminated nineteen residue oligonucleotide of tRNA(Tyr II)(E. coli). PMID- 11945534 TI - Comparative amino acid contents of purified beta-lactamases from enteric bacteria. PMID- 11945535 TI - Biosynthetic relation between the soluble and insoluble dextrans produced by Leuconostoc mesenteroides NRRL B-1299. PMID- 11945536 TI - Tissue specificity of non-histone chromatin phosphoproteins. PMID- 11945537 TI - Concomitance of hog spleen acid DNase and phosphodiesterase activities. PMID- 11945538 TI - Effect of dibutyryl cAMP on the enzymes of fatty acid synthesis and of glycogen metabolism. PMID- 11945539 TI - The metabolism of paraffins in rats. PMID- 11945540 TI - Immunological differences between inducible and constitutive xanthine dehydrogenases in Aspergillus nidulans. PMID- 11945541 TI - ? AB - Presqualenyl-pyrophosphate (PSPP) was synthesized from farnesyl-pyrophosphate (FPP). The purified product could be transformed into a squalene in 90% yield. The structure of PSPP was established by the following experiments:1) PSPP displays a remarkably acid-labile phosphate-ester bond. Analogous to FPP it was possible to stabilize this bond by the addition of bromine. This indicates a structure of the allylester type.2) On acid hydrolysis PSPP splits off not only the pyrophosphate but also water. This may be best explained by the presence of a tertiary hydroxyl group in PSPP.3) The pyrophosphate group of PSPP could be split by prostatic phosphatase. In gas-liquid-chromatography the trimethyl-silylated product gave a single peak. From the mass spectra of this compound and of the corresponding alcohol the expected molecular weight of 426 could be verified for the latter.4) Determining the 3H/(14)C ratio in 1-(3)H(2)-4(14)C-FPP and in PSPP and squalene, synthesized from it, only three 3H-atoms could be detected in PSPP and in squalene.5) Ozonisation of PSPP, synthesized from 1-(14)C-FPP, gave radioactive malondialdehyde, which was characterised as azobenzene malondialdehyde and 4-azobenzene-1-phenyl-pyrazole. The experimental results are in agreement with the following structure of 2, 6, 10, 15, 19, 23-hexamethyl-n tetracosa-2, 6, 11, 14, 18, 22-hexaen-10-yl-pyrophosphate for PSPP. PMID- 11945542 TI - The biosynthesis of plasmalogens in a cell-free system. PMID- 11945543 TI - The subunit molecular weight of acetylcholinesterase. PMID- 11945544 TI - The reaction of N-ethyl maleimide with malate dehydrogenase. PMID- 11945545 TI - The influence of progesterone on the lactate dehydrogenase isoenzymes of the rat reproductive tract. PMID- 11945547 TI - The control of glycogen metabolism in the liver. PMID- 11945546 TI - Erratum. PMID- 11945548 TI - The biology of oncogenic viruses A report on the 2nd Lepetit Colloquium held at the Pasteur, Institute, Paris, during 6-8 November 1970. PMID- 11945549 TI - Aminoacylation of methoxyamine modified tyrosine transfer RNA. PMID- 11945550 TI - A glycogen-debranching enzyme from Cytophaga. PMID- 11945551 TI - A revision of the Meyer-Bernfeld model of glycogen and amylopectin. PMID- 11945552 TI - Uncoupling of electron transport from oxygenation in the mono-oxygenase, orcinol hydroxylase. PMID- 11945553 TI - Methylated oligonucleotides from HeLa cell ribosomal and nucleolar RNA. PMID- 11945554 TI - A proton magnetic resonance study of the interaction between copper and azurin. PMID- 11945555 TI - The effect of cobalt on the synthesis of globin by rabbit reticulocytes in the presence and absence of iron. PMID- 11945556 TI - Soluble acidic lipoprotein components of adrenomedullary chromaffin granules. Relation to chromogranins. PMID- 11945557 TI - Glycoprotein components, sialic acid and hexosamines, bound to inner and outer mitochondrial membranes. PMID- 11945559 TI - Carnitine octanoyltransferase. Evidence for a new enzyme in mitochondria. PMID- 11945558 TI - Small sized ribosomes from mitochondria of locusta migratoria. PMID- 11945560 TI - Reconstitution of methionine-acceptor activity from fragments of E. coli tRNA(fMet) with pCpGp deleted from the 5'-terminus. PMID- 11945561 TI - The hybridization properties of 5'-end sequences of giant nuclear dRNA. PMID- 11945562 TI - Aflatoxin B(1): Cytotoxic mode of action evaluated by mammalian cell cultures. AB - Aflaxton B(1) rapidly inhibits RNA synthesis in rat liver cells, slices or liver in vivo. Established human cells lines (kidney T-cells, HeLa S(3), Chang liver) and mouse fibroblast 3t3 are more slowly affected. Prolonged exposure of synchronized cell cultures to the agent show that cells are retarded in their passage through the S-phase and exhibit a decreased rate of DNA synthesis. Consequent to this, mitosis is also inhibited. Liver cells appear to convert aflatoxin B(1) to a more potent cytotoxin which can then affect normally non susceptible cells. This may explain the susceptibility of liver to tumorogenesis by this carcinogen. PMID- 11945563 TI - Separation and characterization of the constituents of troponin. PMID- 11945565 TI - Stimulation of haemoglobin synthesis in reticulocyte lysates by initiation factors. PMID- 11945564 TI - Growth phase and rotenone sensitivity in Torulopsis utilis: Difference between exponential and stationary phase. PMID- 11945566 TI - Requirement of two protein fractions for O-demethylase activity in Pseudomonas testosteroni. PMID- 11945567 TI - Determination of the amount of DNA on nitrocellulose mebrane filters. PMID- 11945568 TI - Phase transitions and bilayer structure of Mycoplasma laidlawii B. PMID- 11945569 TI - Insoluble hexokinase in the brush border region of rat intestinal epithelial cells. PMID- 11945570 TI - Peptide fragments released from the collagen molecule by pronase. PMID- 11945571 TI - Erratum. PMID- 11945572 TI - Primary structure of aplysia myoglobin: Sequence of a 63-residue fragment. PMID- 11945574 TI - Subunit weight and N-terminal groups of liver and kidney carboxylesterases (EC 3.1.1.1). PMID- 11945573 TI - Reactivity of carboxyl groups in modified proteins. AB - The reactivities of carboxyl groups to carbodiimides in non-reduced, S carboxymethyl, and S-cyanoethyl proteins have been compared. When disulphide bonds are split, a decrease in reactivity was found, relatively minor in the S carboxymethyl derivative and almost complete loss of reactivities in the S cyanoethyl derivative. PMID- 11945575 TI - The primary structure of cytoplasmatic aspartate aminotransferase from pig heart muscle tryptic hydrolysis products. PMID- 11945577 TI - Membrane association of circular forms of ColE(1) DNA: Electron microscopic examination. PMID- 11945576 TI - The dependence of phosphofructokinase kinetics upon protein concentration. PMID- 11945578 TI - 4,5,6,7-Tetrafluorotryptophan: A substrate for tryptophan-tRNA ligase. PMID- 11945579 TI - Intracellular localization of adenyl cyclase and of binding sites for 3', 5' adenosine monophosphate in adrenal cortex. PMID- 11945580 TI - Effects of tannic acid and 2-thenoyltrifluoroacetone on succinate oxidation and penetration across the mitochondrial membrane. PMID- 11945581 TI - The alkyl and alk-1-enyl glycerols in the liver of rats fed long-chain alcohols or alkyl glycerols. AB - Dietary long-chain alcohols and alkyl glycerols, including polyunsaturated compounds, are incorporated into the alkyl and alk-1-enyl moieties of the ionic alkoxylipids of rat liver, whereas polyunsaturated fatty acids are not. PMID- 11945582 TI - Effects of various thyroid states on the metabolism of adenine nucleotides and of glycogen in rat liver. PMID- 11945583 TI - Fluorescence transfer in glutaraldehyde fixed particles of the red alga Porphyridium cruentum (N). PMID- 11945584 TI - The enzymic conversion of squalene, 2(3),22(23)-diepoxide to alpha-onocerin by a cell-free extract of Ononis spinosa. PMID- 11945585 TI - The metabolism of cholesterol by the echinoderms Aterias rubens and Solaster papposus. PMID- 11945586 TI - Species-specific interaction of growth hormones with erythrocyte membranes. PMID- 11945587 TI - Errata. PMID- 11945588 TI - Molecular weight of tomato bushy stunt virus-RNA. PMID- 11945589 TI - Biological activity of ethidium bromide - transfer RNA complexes. PMID- 11945590 TI - Functional adaptation of tRNAs to protein biosynthesis in a highly differentiated cell system. III. Induction of isoacceptor tRNAs during the secretion of fibroin in the silkgland of Bombyx mori L. PMID- 11945591 TI - Preferential binding of E. coli RNA-polymerase to A-T rich sequences of bacteriophage lambda DNA. PMID- 11945593 TI - On the state of copper in the blue protein umecyanin. PMID- 11945592 TI - Conformation of the tRNA minor constituent dihydrouridine. PMID- 11945594 TI - Inhibition of glucagon-stimulated adenyl cyclase by insulin. PMID- 11945595 TI - Activation of calcium efflux by ADP and inorganic phosphate. PMID- 11945596 TI - Calcium efflux dependent formation of ATP from ADP and orthophosphate by the membranes of the sarcoplasmic vesicles. PMID- 11945597 TI - ATP synthesis by the reverse of the sarcoplasmic calcium pump. PMID- 11945598 TI - Species differences in the in vitro metabolism of aflatoxin B(1). PMID- 11945599 TI - Mutants of Aspergillus nidulans lacking malate synthase. PMID- 11945600 TI - pH-Dependent changes in the spin state of cytochrome P450 from adrenal mitochondria. PMID- 11945601 TI - The transport of pyruvate in rat liver mitochondria. PMID- 11945603 TI - The regulation of ribose-5-phosphate isomerisation in Escherichia coli K12. PMID- 11945602 TI - On the use of glutamate dehydrogenase as a mitochondrial marker enzyme for the determination of the intracellular distribution of rat liver pyruvate carboxylase. PMID- 11945604 TI - Induction of lymphocyte transformation by periodate. PMID- 11945605 TI - A new form of mammalian DNA-dependent RNA polymerase and its relationship to the known forms of the enzyme. PMID- 11945607 TI - Differential thermal stability of old and new ribosomal RNA of rat liver. PMID- 11945606 TI - Purification and properties of methionyl-tRNA synthetase from E. coli K 12 carrying the F32 episome. PMID- 11945608 TI - Circular dichroism of mitochondrial ribosomal RNA from Trichoderma viride. PMID- 11945609 TI - The aminoterminal sequence of Dendrostomum pyroides hemerythrin. PMID- 11945610 TI - Proteins of bovine retinal outer segments: Electrophoresis on polyacrylamide gels in the presence of sodium dodecyl sulfate. PMID- 11945612 TI - Structural change of human red cell membranes in the glucose-preloaded state. PMID- 11945611 TI - A study of a tyrosine aminotransferase inactivating system in rat liver homogenate. PMID- 11945613 TI - Role modeste du citrate comme transporteur d'acetyl-CoA chez l'animal vivant. AB - 2,4-(14) C-Citrate incorporated to a far greater extent than 1,5-(14) C-citrate into liver, carcass or adipose tissue fatty acids of living mice. This finding excludes the possibility that the acetyl units emerge from the mitochondria in the form of citrate. PMID- 11945614 TI - Different antigenic determinants in the polypeptide chains of human collagen. PMID- 11945615 TI - On the complex formation between basic pancreatic trypsin inhibitor and TLCK-, TPCK-derivatives of beta-trypsin. AB - The study of the interaction of the pancreatic inhibitor with different alkylated derivatives of alpha- and beta-trypsin shows that: 1) TLCK-beta-trypsin forms a complex with pancreatic inhibitor in tris buffer and tris-ethanol 40% system. 2) TLCK-alpha-trypsin and TLCK-TPCK-beta-trypsin have lost their ability to complex formation with pancreatic inhibitor. TLCK-alpha-trypsin and TLCK-TPCK-beta trypsin are in derivatives in which the "chymotryptic" active site is destroyed. The results presented in this paper prove the participation of the "chymotrypic" active site in the interaction between trypsin and pancreatic inhibitor. This is the second interaction beside that of the electrostatic bond between Asp-117 of trypsin and Lys-15 of the inhibitor which we proved earlier. PMID- 11945616 TI - Amino acid sequence of a fragment of rabbit muscle aldolase. PMID- 11945617 TI - Errata. PMID- 11945618 TI - Dual function and common identity of proteins in glycogen metabolism: A hypothesis. PMID- 11945619 TI - The changes of metabolite content and the control of phosphofructokinase in rat liver in different dietary and hormonal conditions. PMID- 11945620 TI - Effect of amino acid starvation on the degradation of DNA in Escherichia coli B/r after UV irradiation. PMID- 11945621 TI - Nuclease inhibitor from the nuclear SAP of liver and hepatoma cells. PMID- 11945622 TI - Gas-liquid chromatography of hexosamines. PMID- 11945623 TI - Detection of biochemical intermediates by mass fragmentography: Mescaline and tetrahydroisoquinoline precursors. PMID- 11945624 TI - Interactions between ADP and atractyloside on the mitochondrial adenine nucleotide carrier. PMID- 11945625 TI - Some parameters affecting respiratory control in Azotobacter vinelandii membranes. AB - RCI :respiratory control index (state 3/state 4 rate of oxygen uptake)m-C1 CCP: carbonylcyanide m-chlorophenyl hydrazoneHQNO : 2-n-heptyl-4-hydroxyquinoline-N oxidePIPES: piperazine-N,N'-bis 2-ethane sulphonic acid PMID- 11945626 TI - A coupling factor from chromatium strain D chromatophores. PMID- 11945627 TI - Direct determination of DeltapH in chloroplasts, and its relation to the mechanisms of photoinduced reactions. PMID- 11945628 TI - The source of lactate dehydrogenase in preimplantation mouse embryos. PMID- 11945629 TI - Stimulation of hydroxymethylglutaryl-coenzyme a reductase in mouse liver by X rays. PMID- 11945630 TI - Electrophoretic analysis of hydrolases from grown gall tissues. PMID- 11945631 TI - Tetrameric structure of nucleoside diphosphate kinase from pea seed. PMID- 11945632 TI - Opposite effects of potassium ions on the affinities of rat liver serine dehydratase for coenzyme and substrate. PMID- 11945633 TI - Growth hormone stimulation of protein synthetic activity of membrane-bound ribosomes. PMID- 11945634 TI - Peptidyl puromycin synthesis; effect of several antibiotics which act on 50 S ribosomal subunits. AB - The effects of several antibiotics which are know to bind with 50 S ribosomal subunits, on the formation of several di- and tri-peptidyl puromycins have been examined. Tylosin and spiramycin inhibited the formation of phenylalanyl-(14)C phenylalanyl-puromycin, glycyl-(14)C-phenyllalanyl-puromycin, leucyl-(14)C phenylalanyl-puromycin, N(epsilon)-carbobenzoxylysyl-(14)C-phenylalanyl puromycin, and valyl-glycyl-(14)C-phenylalanyl-puromycin as well as N-acetyl (14)C-phenylalanyl-puromycin. Of these compounds, erythromycin and oleandomycin selectively inhibited the formation of phenylalanyl-(14)C-phenylalanyl-puromycin. Although chloramphenicol and lincomycin inhibited the formation of most of these peptidyl puromycins, the formation of phenylalanyl-(14)C-phenylalanyl-puromycin and leucyl-(14)C-phenylalanyl-puromycin was found to be resistant to these antibiotics. So far, no significant effect of siomycin has been observed on pepetidyl puromycin formation in the absence of G factor. PMID- 11945636 TI - Mitochondrial ribosomes. PMID- 11945635 TI - Lysosomal hydrolases: Conversion of acidic to basic forms by neuraminidase. PMID- 11945637 TI - Inhibitory effects of fluorocitrates on yeast mitochondria. PMID- 11945638 TI - Kinetics of metal chelatase from rat liver mitochondria. PMID- 11945639 TI - Implication of SH-groups in the mitochondrial energy-coupling system revealed by measurements of 14C-ethacrynate incorporation into rat liver mitochondria. PMID- 11945641 TI - Evidence for a non-helical region at the carboxyl terminus of the collagen molecule. PMID- 11945640 TI - The relationship between P 700 and NADP reduction in chloroplasts. PMID- 11945642 TI - Studies on the bacteriophage MS2. Nucleotide fragments from the coat protein cistron. PMID- 11945643 TI - Isolation of a tyrosine-tRNA-tsDNA hybrid. PMID- 11945644 TI - Concurrent measurements of adenosine 3', 5'-monophosphate, tyrosine aminotransferase and alpha-aminoisobutyrate uptake in rat liver after injection of glucagon. PMID- 11945645 TI - Immunochemical studies of the O-antigens of Vibrio cholerae. The constitution of a lipopolysaccharide from V. cholerae 569B (Inaba). PMID- 11945647 TI - Relationship between light-induced quenching of atebrin fluorescence and ATP formation in Rhodospirillum Rubrum chromatophores. PMID- 11945646 TI - RNA Directed DNA synthesis: Identification in L5178Y mouse leukemic cells and distribution of the polymerase in a synchronized L5178Y cell population. PMID- 11945648 TI - Pathway of fructose utilization by Escherichia coli. PMID- 11945649 TI - Erratum. PMID- 11945650 TI - Isolation and properties of a regulatory mutant in the hexose phosphate transport system of Escherichia coli. PMID- 11945651 TI - ? AB - Three double mutants were prepared from single mutant chl-r strains of Escherichia coli K12 lacking nitrate reductase activity. When extracts of these three double mutants were mixed, nitrate reductase activity was reconstituted. PMID- 11945652 TI - Diminution in starvation of capacity of cell sap to support protein synthesis in cell-free systems. PMID- 11945653 TI - Consequences of heme loss in unstable hemoglobins: A study of hemoglobin koln. PMID- 11945654 TI - ATP, calcium uptake and growth hormone release. PMID- 11945655 TI - A dynamic molecular model for transfer RNA. PMID- 11945656 TI - Order of substrate binding to tyrosyl-tRNA synthetase of Escherichia coli B. PMID- 11945657 TI - Effect of the extent of DNA transcription of plant cells and bacteria on the transcription in plant cells of DNA released from bacteria. PMID- 11945658 TI - Binding of termination factor RHO to RNA polymerase and DNA. PMID- 11945659 TI - RNA Polymerase activity in purified nuclei from rat prostate gland in the presence of polyamines. PMID- 11945660 TI - Inhibitors of purine biosynthesis. PMID- 11945661 TI - Interaction of the apoenzyme of L-glutamate decarboxylase with pyridoxal phosphate analogues. PMID- 11945662 TI - Molecular parameters of the beef heart succinate dehydrogenase. PMID- 11945663 TI - Cyclic 3', 5'-amp in Saccharomyces carlsbergensis under various conditions of catabolite repression. PMID- 11945665 TI - Resolution of a cross-reacting anti-human growth hormone serum into a monospecific antibody population. PMID- 11945664 TI - Correlations between thermal stability and circular dichroism of hemoglobin derivatives of different species. PMID- 11945666 TI - Formation of long-chain alcohols in the gastro-intestinal tract of the rat. PMID- 11945667 TI - Interconversion of inactive to active pyruvate dehydrogenase in rat liver after fructose application in vivo. PMID- 11945668 TI - The heterogeneity of the non-histone chromatin proteins from mouse tissues. PMID- 11945669 TI - Purification of the lactose specific factor III of the staphylococcal PEP dependent phosphotransferase system. PMID- 11945670 TI - The regulation of Escherichia coli methylglyoxal synthase; a new control site in glycolysis? PMID- 11945671 TI - Glyceraldehyde-3-phosphate dehydrogenase from Bacillus stearothermophilus. PMID- 11945672 TI - The reduction of cytochrome c by hydrated electrons. PMID- 11945673 TI - The composition of cyanogen bromide cleavage products from hen's egg phosvitin. PMID- 11945674 TI - Synthesis of hamamelose-diphosphate by isolated spinach chloroplasts. PMID- 11945675 TI - Regulation of pig heart mitochondrial glutamate dehydrogenase by nucleotides and phosphate: Comparison with pig heart and beef liver purified enzymes. PMID- 11945676 TI - Mechanism of mutagenesis by N-methyl-N'-nitro-N-nitrosoguanidine (MNNG). Methylation of nucleic acids by N-trideuteriomethyl-N'-nitro-N-nitrosoguanidine (D3-MNNG) in the presence of cysteine and in cells of Escherichia coli. PMID- 11945677 TI - Polyribosomes of E. coli: The distribution of free 70 S particles and subunits in the presence of K(+) and Na(+) ions. PMID- 11945678 TI - Substrate and antibiotic binding sites at the peptidyl transferase centre of E. coli ribosomes: Binding of UACCA-Leu to 50 S subunits. PMID- 11945679 TI - Respiration-dependent anion uptake by rat liver mitochondria. PMID- 11945680 TI - Effects of uncoupler on rat liver mitochondria isolated after chloramphenicol treatment. PMID- 11945681 TI - The sites of cobra venom phospholipase action on the respiratory system of submitochondrial particles. PMID- 11945682 TI - Succinate oxidase activity in the absence of ubiquinone. PMID- 11945683 TI - Effect of siomycin on the G factor dependent GTP hydrolysis by Escherichia coli ribosomes. PMID- 11945684 TI - DNA-dependent RNA polymerase from vegetative cells and from spores of Bacillus subtilis. IV. Subunit composition. PMID- 11945685 TI - Studies on the origin of the form Ib mammalian DNA-dependent RNA polymerase. PMID- 11945686 TI - Involvement of the RC locus in RNA synthesis inhibition by levallorphan in E. coli. PMID- 11945687 TI - ? AB - Some rat liver fatty acid synthetase has been isolated from microsomal fraction. Enzyme activity and electro micrography of purified enzyme show that the microsomal enzyme was the same as that of the supernatant. These results are discussed in relation to the role of microsomes in the de novo synthesis of fatty acids synthesis in the cell. PMID- 11945688 TI - ? AB - Mice were given glucose-U-(14)C and the radioactivity of triglyceride and phospholipid glycerol and fatty acids was measured 30 sec, 3 or 30 min after the i.v. administration in liver microsomes and supernatant. After the acetate administration the radioactivity of palmitic and stearic acids and that of their carboxyl carbons was also measured. The repartition of the radioactivity was discussed in relation with the localization of fatty acid synthesis. In agreement with our previous results it was suggested that the endoplasmic reticulum is the main site of this synthesis. PMID- 11945689 TI - A unique electrophoretic pattern of triosephosphate isomerase in human cultured fibroblasts. PMID- 11945690 TI - 5-(2-Thiolyl)-1,5-dihydro-flavin: A model of group transfer in flavin dependent substrate dehydrogenation. PMID- 11945691 TI - Localization of invertase in yeast vacuoles. PMID- 11945692 TI - The carboxy-terminal primary structure of the alpha subunit from bovine and porcine luteinizing hormone. PMID- 11945693 TI - Organ specificity of histone Acetyltransferases. PMID- 11945694 TI - Enzymatic determination of the unit chain length of glycogen and related polysaccharides. PMID- 11945695 TI - Resolution of chloroplast lamellar proteins by electrophoresis in polyacrylamide gels. Different patterns obtained with fractions enriched in either chlorophyll a or chlorophyll b. PMID- 11945696 TI - Plastocyanin-dependent photoreduction of NADP by agranal chloroplasts from maize. PMID- 11945697 TI - The properties of free and membrane bound polysomes studied in a cell free system of Bombix mori silkglands. PMID- 11945699 TI - Conformational differences between the native and denatured forms of tRNA(Ser) and tRNA(Phe) from yeast. PMID- 11945698 TI - The 3'-terminal nucleotide sequence of the 16 S ribosomal RNA from Escherichia coli. PMID- 11945700 TI - 5-methyl-2-thiouridine: A new sulfur-containing minor constituent from rat liver glutamic acid and lysine tRNAs. PMID- 11945701 TI - Stoichiometry of the interaction of lima bean protease inhibitor with trypsin and/or chymotrypsin. PMID- 11945702 TI - Mechanism of TSH action. Studies with dibutyryl cyclic AMP and dibutyryl cyclic GMP. PMID- 11945704 TI - Nucleic acid-protein interactions A Report on the Third Miami Winter Symposium held in Miami, Florida 18-19 January 1971. PMID- 11945703 TI - Activation of polynucleotide phosphorylase by 3',5'-cyclic AMP, ATP-dependent protein kinase. PMID- 11945705 TI - Chromatography of proteins and peptides on sephadex ion-exchangers: Dependence of the resolution on the elution schedule. PMID- 11945706 TI - The effects of 2,3-diphosphoglycerate on native and chemically modified horse hemoglobin. PMID- 11945707 TI - Optical rotatory dispersion and circular dichroism of baker's yeast transketolase in the far ultraviolet region of the spectrum. PMID- 11945708 TI - Specific and non-specific binding of an organophosphate spin label to some serine esterases. PMID- 11945709 TI - Two different forms of pyruvate kinase in rat kidney cortex. PMID- 11945710 TI - Purification and characterization of wheat germ hexokinases. PMID- 11945711 TI - Effect of spermine on free nucleotide turnover in chick embryo. PMID- 11945712 TI - Crosslinking of human fibrin: Evidence for intermolecular crosslinking involving alpha-chains. PMID- 11945713 TI - Preparation and isolation of the S-carboxymethyl derivative chains of human fibrinogen. PMID- 11945715 TI - The effect of gougerotin analogues on ribosomal peptidyl transferase. PMID- 11945714 TI - Inhibitory effect of naturally occurring and chemically modified amatoxins on RNA polymerase of rat liver nuclei. PMID- 11945716 TI - Regulation of pteroylglutamate pool size by methionine in Saccharomyces cerevisiae. PMID- 11945717 TI - Restoration of electron transport in ultraviolet-irradiated membranes of Aerobacter aerogenes. PMID- 11945718 TI - ? AB - A cell-free preparation was obtained from a culture of Mycobacterium avium. This preparation contained an L-alanine reductase which reduces L-alanine to L alaninol, a constituant of the mycoside C(2). This reduction is better when ATP and NADH are present in the incubation medium. PMID- 11945719 TI - The approximate red absorption band of chlorophyll b in Ulva lactuca at 77 degrees K. PMID- 11945721 TI - Dependence on Mg(2+) for different states of the membrane-bound adenosine triphosphatase of Micrococcus lysodeikticus. PMID- 11945720 TI - L-serine transport in membrane vesicles of Bacillus subtilis energized by NADH or reduced phenazine methosulfate. PMID- 11945722 TI - Joint Biochemical Meeting of the Societe Belge de Biochimie - Belgische Vereniging voor Biochemie and the Gesellschaft fur Biologische Chemie on Peptides and Proteins and Antigens and Immunogenicity held in Liege on January 14-16, 1971. PMID- 11945723 TI - Protein A from Staphylococcus aureus conversion of complement factor C3 by aggregates between IgG and protein A. PMID- 11945724 TI - Protein A from Staphylococcus aureus complement fixation by aggregates between protein A and IgG1 or IgG2 from guinea pig. PMID- 11945726 TI - The effect of cations and complexing agents on the hydrolysis of L-Alanine-4 nitroanilide by aminopeptidase M. PMID- 11945725 TI - Succinylation of the amino group of isoleucine 16 in delta-chymotrypsin with retention of activity. PMID- 11945727 TI - On the mechanism of amide bond cleavage catalyzed by aminopeptidase M. Kinetic studies in deuteriumoxide. PMID- 11945728 TI - Enzyme entrapment in liposomes. PMID- 11945729 TI - The turnip lysozyme. PMID- 11945731 TI - The substrate specificity of L-asparaginase from Alcaligenes eutrophus. PMID- 11945730 TI - Coordinative changes in the activities of enzymes in carbohydrate metabolism during oogenesis in Mysgurnus fossilis. PMID- 11945732 TI - Participation of a lipid intermediate in the biosynthesis of Saccharomyces cerevisiae LK2G12 mannan. PMID- 11945733 TI - Effect of non-ionic agents on the stability of association of ribosomal subparticles. PMID- 11945734 TI - ESR of Cu(2+) bound to polynucleotides and derivatives. AB - Binding of Cu(2+) to polynucleotides has been found to occur at 1 Cu/5 polymer P and at similar molar ratios of copper to selected nucleic acid derivatives. Electron spin resonance (ESR) measurements show that the g-values and the hyperfine splitting constants vary substantially among the different complexes. Binding of Cu(2+) to the phosphates as well as to the bases is observed, and in some cases occurs simultaneously. PMID- 11945735 TI - Incorporation of deoxyribonucleoside triphosphates into plasmid DNA by toluenized cells of thermosensitive DNA replication mutants of Escherichia coli. PMID- 11945737 TI - Changes of aminoacyl-tRNA synthesis in the liver of rats administered cholesteryl 14-methylhexadecanoate. PMID- 11945736 TI - The superhelix density of bacteriophage PM2 DNA, determined by a viscometric method. PMID- 11945738 TI - Virus-specific informosome components in the extracts of newcastle disease virus infected cells. PMID- 11945739 TI - The distribution of virus-specific RNA between informosome component and polyribosomes in the extract of NDV-infected cells. PMID- 11945740 TI - The binding sites for Escherichia coli RNA polymerase on lambda phage DNA: Characteristics of binding sites on sonicated lambda DNA. PMID- 11945741 TI - Halobacterium cutirubrum RNA polymerase: Subunit composition and salt-dependent template specificity. PMID- 11945742 TI - The affinity of fluorophenylalanines to phenylalanine-tRNA ligases from Escherichia coli and baker's yeast. PMID- 11945744 TI - Inhibition of photophosphorylation by beta-bromo-beta-nitrostyrene. PMID- 11945743 TI - Template activity of brominated DNA for DNA polymerase. PMID- 11945745 TI - Highly polymerous nuclear DNA from calf thymus isolated by gel filtration in 4 M guanidine hydrochloride. PMID- 11945746 TI - Circular dichroism and association-dissociation of ribosomes. PMID- 11945747 TI - The binding of polysomes to smooth membranes of rat liver promoted by steroid hormones and extracts from either rough endoplasmic reticulum or from polysomes of the opposite sex. PMID- 11945748 TI - Comparison on effects of phenobarbital and nicotine on nuclear protein synthesis in rat liver. PMID- 11945749 TI - Enzymic synthesis of loganin by carboxyl group methylation of loganic acid. PMID- 11945750 TI - Two-component inhibition as a method for studying of enzyme active centre flexibility. Co-inhibition of tryptic activity by Ag(+) and Pb(2+) ions. PMID- 11945751 TI - Reaction of arginine residues in basic pancreatic trypsin inhibitor with phenylglyoxal. PMID- 11945752 TI - Some modifications of the kinetic properties of bovine liver glutamate dehydrogenase (NAD(P)) covalently bound to a solid matrix of collagen. PMID- 11945753 TI - Identification of the secondary structure of polypeptide chains in solution by x ray diffusion scattering. PMID- 11945754 TI - Errata. PMID- 11945755 TI - Dual function and common identity of proteins in glycogen metabolism: Reply to a hypothesis. PMID- 11945756 TI - Acceptor activity of valine tRNA modified with CME-carbodiimide. Heterogeneity of modified tRNA. PMID- 11945757 TI - Reaction of a spin-labelled carbodiimide with nucleosides, poly U and tRNA. PMID- 11945759 TI - Changes of stability and conformation of DNA following the covalent binding of a carcinogen. PMID- 11945758 TI - Electron microscopy study of 70 S ribosomes of Escherichia coli. PMID- 11945760 TI - Origin of the ribosome specific factors responsible for peptide chain elongation in yeast. PMID- 11945761 TI - The mitochondrial ribosome and ribosomal RNA of the chick. PMID- 11945762 TI - Evidence for the absence of a central core in particles aggregated from protein subunits of bacteriophage fr. PMID- 11945764 TI - Synthesis by ribonuclease a of codons containing modified nucleosides in the 'wobble' position. PMID- 11945763 TI - Comparison of the amino terminal sequences of bovine, dogfish, and lungfish trypsinogens. PMID- 11945765 TI - The presence of plasma membrane enzymes on the surface of bovine milk fat globules. PMID- 11945766 TI - On the mechanism of the energy-dependent quenching of atebrin fluorescence in isolated chloroplasts. PMID- 11945767 TI - Heterogeneity of PIG immunoglobulin gamma-chains. PMID- 11945768 TI - Photosynthetic control in broken spinach chloroplasts. PMID- 11945770 TI - Subunit size of transketolase from baker's yeast. PMID- 11945769 TI - ? AB - Two auxin oxidases with distinct antigenic specificities have been identified in extracts of plant tumor induced in Datura stramonium by Agrobacterium tumefaciens. Auxin oxidase and peroxidase activities are demonstrated for both enzymes. The adaptation of an immunochemical method allows the quantitative evaluation of both enzymes in different plant tissue extracts. The amount of these enzymes which is low in the organ of the healthy plant increases during tumorisation, healing and in vitro culture of healthy tissues. PMID- 11945771 TI - A thermochemical study of the reaction between protein A from S. aureus and fragment Fc from immunoglobulin G. PMID- 11945772 TI - ? AB - In the extracts of Quercus pedunculata roots, the activities of 5-dehydroquinate hydro-lyase and shikimate: NADP oxydoreductase are eluted in the same fractions whatever separation procedures are used: chromatography on Sephadex G 100, DEAE cellulose and hydroxylapatite, centrifugation in sucrose density gradient, disc electrophoresis. These results are closely related to those obtained on the aggregate of enzymes involved in the shikimate pathway in Neurospora crassa. The molecular weight of the association determined by gel filtration is about 55,000. PMID- 11945773 TI - Spectroscopic evidence for cis-[PtCl(2)(NH(3))(2)]-erythrocyte membrane interaction. PMID- 11945775 TI - Isolation and study of functional membrane proteins Present status and future prospects. PMID- 11945774 TI - Phosphorylation coupled to non-cyclic electron flow in photosystem I. PMID- 11945776 TI - Contaminant of malic acid: A malic enzyme inhibitor. PMID- 11945777 TI - Interaction of transferase II with the 60 s ribosomal subunit. PMID- 11945778 TI - Polyribosome formation from haemoglobin ribonucleoprotein in vitro. PMID- 11945780 TI - A study of tRNA methylases by the dissected molecule method. PMID- 11945779 TI - R-factor mediated gentamicin resistance: A new enzyme which modifies aminoglycoside antibiotics. PMID- 11945781 TI - 4Phe-(6)Val-antamanide, an antitoxic cyclodecapeptide with C(2) symmetry. PMID- 11945782 TI - Endogenous phosphate-dependent phosphorylation of ADP and oligomycin-sensitive ADP-ATP exchange A quantitative assessment of their activity in inner membrane matrix particles from rat liver mitochondria. PMID- 11945783 TI - The effect of 2-phenylisatogen on oxidative phosphorylation in rat liver mitochondria. PMID- 11945785 TI - Affinity chromatography of enzymes on insolubilized cofactors. PMID- 11945784 TI - Transport of phosphoenolpyruvate by the tricarboxylate transporting system in mammalian mitochondria. PMID- 11945786 TI - Tissue-specific isoenzyme patterns of creatine kinase (2.7.3.2.) in trout. PMID- 11945787 TI - Studies on the protoplast-bursting factor from Bacillus amyloliquefaciens. PMID- 11945788 TI - Calcium requirements in the action of thyrotropin on the thyroid. PMID- 11945789 TI - The role of phosphoglycerate kinase in the metabolism of Pseudomonas putida. PMID- 11945790 TI - Arginine metabolism in Chlamydomonas reinhardi. Regulation of uptake and breakdown. PMID- 11945791 TI - A new procedure for the isolation and fractionation of histones. PMID- 11945792 TI - Purification and characterization of the cysteine-containing F3 histone from chicken erythrocyte. PMID- 11945793 TI - Concerning the subunit structure of polynucleotide phosphorylase from E. coli. PMID- 11945794 TI - Conformational transition of the synthetic 2-15 N-terminal fragment of hen egg white lysozyme. PMID- 11945795 TI - On the equilibrium of the association-dissociation reaction of ribosomal subparticles and on the existance of the so-called '60 S intermediate' ('swollen 70 S') during centrifugation of the equilibrium mixture. PMID- 11945796 TI - Base sequence distribution in Tetrahymena mitochondrial DNA. PMID- 11945797 TI - ? AB - A component with high molecular weight is prepared from commercial bovine insulin by gel filtration on Sephadex G 50. The substance has little biological and immunological activity in comparison to insulin. The activity is not changed by incubation with trypsin. It associates in 1 M CH(3)COOH and possesses a molecular weight of 28,500 as shown by ultracentrifugation. The ability of the component to precipitate with bovine insulin antibodies demonstrates that it contains immunologically active insulin-like proteins and is not a homogenous substance. PMID- 11945799 TI - Errata. PMID- 11945798 TI - Role of fructose-1,6-diphosphatase in fructose utilization by Escherichia coli. PMID- 11945800 TI - Plant hormones: Metabolism and interaction A report of the FEBS Summer School no. 12 held in Halle (Saale), German Democratic Republic, on 14-18 November, 1969. PMID- 11945801 TI - The subunit molecular weights of the alpha-ketoacid dehydrogenase multienzyme complexes from E. coli. PMID- 11945802 TI - A structural aspect of human fibrinogen suggested by its plasmin degradation. PMID- 11945803 TI - The interaction of 1-anilino-8-naphthalene sulphonate with yeast alcohol dehydrogenase. PMID- 11945804 TI - On the influence of ATP on the electron paramagnetic resonance spectrum of methemoglobin. PMID- 11945805 TI - Labelling of the active site of beta-galactosidase by N-bromoacetyl beta-D galactopyranosylamine. PMID- 11945807 TI - On the mechanism of enzymatic hydrolysis of carbamyl phosphate. PMID- 11945806 TI - Oxidative deamination of D-homocysteinesulfinic acid. PMID- 11945808 TI - Purification and characterization of human chorionic gonadotropin. PMID- 11945809 TI - Chemical modification of human chorionic gonadotropin and its biological and immunological characterization. PMID- 11945810 TI - Polypeptide chains of intermediate molecular weight in myosin preparations. PMID- 11945812 TI - Studies on the cyanide insensitive oxidase of plant mitochondria. PMID- 11945811 TI - Inhibition of post-UV irradiation growth in the dark of Tetrahymena pyriformis by caffeine and the oncogenic mycotoxin luteoskyrin. PMID- 11945813 TI - Spin state changes in cytochrome P-450 associated with cholesterol side chain cleavage in bovine adrenal cortex mitochondria. PMID- 11945814 TI - Inhibition of proline and lysine hydroxylation prevents normal extrusion of collagen by 3T6 fibroblasts in culture. PMID- 11945815 TI - Rat liver mitochondrial polynucleotide phosphorylase. PMID- 11945816 TI - Fragments of yeast tRNA(Phe) and tRNA(Ser) prepared by partial digestion with spleen phosphodiesterase. PMID- 11945817 TI - Studies on ornithine decarboxylase activity in normal and T2-infected Escherichia coli. PMID- 11945818 TI - Purification of influenza virus by phase separation and selective adsorption. PMID- 11945819 TI - An investigation of the minor base composition of transfer RNA in normal human brain and malignant brain tumors. PMID- 11945820 TI - Permeability properties of rat live endoplasmic reticulum. PMID- 11945821 TI - Energy conservation in the photochemical reactions of photosynthesis and its relation to delayed fluorescence. PMID- 11945822 TI - Inactivation and reactivation of the acceptor binding site of ribosomal peptidyl transferase. PMID- 11945823 TI - The function of pseudouridylic acid in transfer ribonucleic acid. Photochemical and chemical modification of formylmethionine tRNA of E. Coli. PMID- 11945824 TI - The function of pseudouridylic acid in transfer ribonucleic acid. Irradiation and cyanoethylation of E. coli valine tRNA fragments. PMID- 11945825 TI - The partial amino acid sequence of sheep heart myoglobin. PMID- 11945826 TI - Light-initiated enzymic activity caused by photostereoisomerization of cis-4 nitrocinnamoyl-alpha-chymotrypsin. PMID- 11945827 TI - Determination of the individual rate constants of alpha-chymotrypsin-catalyzed hydrolysis with the added nucleophilic agent, 1,4-butanediol. PMID- 11945829 TI - An anionic trypsin-like enzyme from Streptomyces eythreus. PMID- 11945828 TI - On the relationship between structure and reactivity of alpha-chymotrypsin substrates. PMID- 11945830 TI - Spectroscopic evidence for two b cytochromes in mitochondria of intact ehrlich ascites tumor cells. PMID- 11945831 TI - Study of the N-acetylmuramyl-L-alanine amidase activity in Escherichia coli. PMID- 11945832 TI - Purification of three proteins from the outer membrane of the envelope of Escherichia coli. PMID- 11945833 TI - The presence of citrulline in a myelin protein fraction. PMID- 11945834 TI - Nuclear magnetic resonance studies on the interaction between salicylate and ceruloplasmin. PMID- 11945835 TI - Inhibitory effect of a-amanitin on the induction of delta-aminolevulinate synthetase in chick embryo liver. PMID- 11945836 TI - Differences in the vitro amino acid labelling pattern of mitochondria from melanoma and liver. PMID- 11945837 TI - Excretion of 3beta-hydroxy-5-cholenoic and 3a-hydroxy-5a-cholanoic acids in urine of infants with biliary atresia. PMID- 11945839 TI - Structural evidence for two alpha-amanitin sensitive RNA polymerases in calf thymus. PMID- 11945838 TI - Separation of 32P-labelled ribonucleic acid components. The use of polyethylenimine-cellulose (TLC) as a second dimension in separating oligoribonucleotides of '4.5 S' and 5 S from E. coli. PMID- 11945840 TI - Purification of RNA polymerase B activity from rat liver. PMID- 11945841 TI - Purification of the rat liver form B DNA-dependent RNA polymerases. PMID- 11945842 TI - Core particles and proteins from mitochondrial ribosomes of yeast. PMID- 11945844 TI - Dependence of dissociation-association of uncharged ribosomes of Escherichia coli on the Mg(2+) concentration, ionic strength, pH and temperature. PMID- 11945843 TI - Cell propagation on films of polymeric fluorocarbon as a means to regulate pericellular pH and pO(2) in cultured monolayers. PMID- 11945845 TI - Mammalian chain initiation: The effect of aurintricarboxylic acid. PMID- 11945846 TI - Differential activation of lipoprotein lipase from human post-heparin plasma, milk and adipose tissue by polypeptides of human serum Apolipoprotein C. PMID- 11945847 TI - Two b cytochromes of pigeon heart mitochondria. PMID- 11945848 TI - Biosynthesis and properties of 2-thioadenyl cobamide and its coenzyme form. PMID- 11945849 TI - Effect of the secondary structure of globular proteins in solution on the indicatrix of x-ray diffusion scattering. PMID- 11945850 TI - X-ray small-angle scattering on soluble antigen-antibody complexes. PMID- 11945851 TI - Effect of N(6)-(Delta(2)-isopentenyl)-adenosine on thymus and spleen in relation to antibody formation in mice. PMID- 11945852 TI - Control of pyruvate dehydrogenase interconversion in adipose tissue by insulin. PMID- 11945853 TI - Immunoassay using antigen-enzyme conjugates. PMID- 11945854 TI - Mass spectrometric sequence determination of permethylated peptide mixtures. PMID- 11945856 TI - Studies on low-molecular weight proteoglycans isolated from rabbit ear cartilage. PMID- 11945855 TI - The stereochemistry of the abortive transmination shown by glutamate decarboxylase. PMID- 11945857 TI - The effect of erythromycin on peptide bond formation and the termination reaction. PMID- 11945858 TI - The subcellular distribution of colchicine-binding protein ('microtubule protein') in rat brain. PMID- 11945859 TI - Errata. PMID- 11945860 TI - Use of inhibitors to study the structure and function of nucleic acids and ribosomes Recent advances. PMID- 11945861 TI - Fumarate transport by rat liver mitochondria. PMID- 11945863 TI - Inhibition of pigeon liver NAD kinase by a non-phosphorylatible analogue of NAD. PMID- 11945862 TI - Inhibitors of an RNA-dependent DNA polymerase. PMID- 11945864 TI - Incorrect heterologous aminoacylation of various yeast tRNAS catalysed by E. coli valyl-tRNA synthetase. PMID- 11945865 TI - Primary structure of tRNA(val)(2) from brewer's yeast. PMID- 11945866 TI - Sensitivity and resistance to streptomycin in relation with factor-mediated dissociation of ribosomes. PMID- 11945867 TI - Interconversion of pyruvate dehydrogenase in rat heart muscle upon perfusion with fatty acids or ketone bodies. PMID- 11945868 TI - Stimulation with insulin in vitro of protein synthesis by diabetic hepatic microsomes. PMID- 11945869 TI - A general method for distinguishing between endo and exo actions of carbohydrases. PMID- 11945870 TI - Pyrrolooxygenase: A new type of target enzyme. PMID- 11945871 TI - Glycolipids of a halotolerant moderately halophilic bacterium. PMID- 11945872 TI - DNA-methylase in loach embryos (Misgurnus fossilis). PMID- 11945873 TI - EPR studies of nitrogenase: ATP dependent oxidation of fraction 1 protein by cyanide. PMID- 11945874 TI - Studies of the composition and structure of plasma lipoproteins. C- and N terminal amino acids of the two nonidentical polypeptides of human plasma apolipoprotein A. PMID- 11945875 TI - Formation of polynucleate avian erythrocytes by polylysine and phospholipase C. PMID- 11945876 TI - Relationship between cyclic AMP dependent protein kinase(s) and cyclic AMP binding protein(s) in rat liver. PMID- 11945877 TI - Conformational transition of a myelin protein. PMID- 11945878 TI - Rapid isolation and separation of the non-histone proteins of rat liver nuclei. PMID- 11945879 TI - Poly 2'-deoxy-2'-chlorouridylic and -cytidylic acids. PMID- 11945880 TI - Plastoquinone mediates electron transport between cytochrome b-559 and cytochrome f in spinach chloroplasts. PMID- 11945881 TI - A possible regulation of activity of ferredoxin-NADP reductase ferredoxin system by ionic strength: Catalytic significance of the one to one complex. PMID- 11945882 TI - Purification and substrate specificity of lecithin-cholesterol acyl transferase from human plasma. PMID- 11945883 TI - The mitochondrial ribosome from Locusta migratoria: Dissociation into subunits. PMID- 11945884 TI - Separation of enzyme activities catalysing spermidine and spermine synthesis in rat brain. PMID- 11945885 TI - A monospecific antibody population in cross-reacting anti-human placental lactogen serum. PMID- 11945886 TI - Ornithine decarboxylase and ribosomal RNA synthesis during the stimulation of lymphocytes by phytohaemagglutinin. PMID- 11945887 TI - Studies on inhibitors of mammalian tRNA methylases. PMID- 11945888 TI - Comparative agar gel electrophoresis of RNA, ribosomes and ribosomal subunits of higher eukaryotic organisms. PMID- 11945889 TI - Hormonal effects on the rat gonadal lactate dehydrogenases. PMID- 11945890 TI - The molecular weight of a yeast pyruvate kinase and its subunits. PMID- 11945891 TI - Kinetics of the reverse reaction catalyzed by glutathione reductase of yeast. PMID- 11945893 TI - The inhibition of yeast alpha-glucosidase by L-1,2-anhydro-myo-inositol. PMID- 11945892 TI - Change in anomeric form of the product induced by the aglycone alpha-amylase catalyzed hydrolyses of phenyl alpha-maltosides. PMID- 11945894 TI - ? PMID- 11945896 TI - Mechanism of chymotrypsin hydrolysis of an ester and an anilide. PMID- 11945895 TI - Effect of 5-fluorouracil on beta-galactosidase synthesis in an Escherichia coli mutant resistant to catabolite repression of the lac operon. PMID- 11945897 TI - Hydrolysis of uridine-5' N-aryl and N-alkyl phosphoramidates by ribonucleoside-5' phosphoramidase. PMID- 11945898 TI - Glycerol 3-phosphate dehydrogenase: loss of DPNH binding following photooxidation. PMID- 11945899 TI - Partial degradation of 4.7.10.13.16-docosapentaenoic acid in rat liver. PMID- 11945900 TI - Sugar uptake by germinating Bacillus subtilis spores. PMID- 11945901 TI - Involvement of sulfhydryl groups in the activation of latent ribonuclease of rat liver polyribosomes. PMID- 11945902 TI - Isolation and characterization of the large cyanogen bromide peptides from the alpha1- and alpha2-chains of pig skin collagen. PMID- 11945904 TI - Immunoglobulin a biosynthesis. Intracellular accumulation of 7 S subunits. PMID- 11945903 TI - The nucleotide sequence of a serine tRNA from Escherichia coli. PMID- 11945905 TI - Hybrid RNA polymerases formed from core enzymes and sigma factors of E. coli and thermophilic B. megaterium. PMID- 11945906 TI - On the localisation of enzymes of deoxynucleoside catabolism in Escherichia coli. PMID- 11945907 TI - Identification of digalactosylcysteine in a glycopeptide isolated from urine by a new preparative technique. PMID- 11945908 TI - Age related changes in the reducible cross-links of collagen. PMID- 11945909 TI - Evidence for cooperative effects in the oxidation of deoxyhemerythrin by ferricyanide. PMID- 11945910 TI - Localization of the cholinergic receptor protein by immunofluoroscence in eel electroplax. PMID- 11945911 TI - Biosynthesis of 4-aminobenzoic acid in Aerobacter aerogenes. PMID- 11945912 TI - Erythrocyte fragility and potassium efflux as affected by temperature and hemolyzing rate. PMID- 11945914 TI - Studies on the biosynthesis of dermatan sulfate. PMID- 11945913 TI - Deuteron resonance: A novel approach to the study of hydrocarbon chain mobility in membrane systems. PMID- 11945915 TI - Thionucleotide formation in Escherichia coli transfer ribonucleic acid in the presence of 5-fluorouracil. PMID- 11945916 TI - Bacterial RNA synthesis in animal cells following bacterial contact. PMID- 11945917 TI - Influence of polyamines and salts on changing patterns of tRNA methylation. PMID- 11945918 TI - Binding of corticosterone to liver chromatin acidic proteins. PMID- 11945919 TI - Involvement of highly reactive sulfhydryl groups in the action of RNA polymerase from E. coli. PMID- 11945920 TI - The primary structure of a monoclonal lambda-type immunoglobulin L-chain of subgroup II (Bence-Jones protein NEI): Evolutionary origin of antibody variability. PMID- 11945921 TI - Specific aggregation products of histone fractions (presence of cysteine in F2a1 from echinoderms). PMID- 11945922 TI - Endogenous acylhydrolase breaks down diglyceride in 5 degrees phospholipase C treated rat liver microsomes. Products inhibit glucose 6-phosphohydrolase. PMID- 11945924 TI - The reaction rate constant of hydrated electron with some hemoproteins as a function of the pH. PMID- 11945923 TI - Degradation of phenolic compounds in plant cell cultures. PMID- 11945925 TI - Errata and addenda. PMID- 11945927 TI - Cyclic adenosine-3', 5'-monophosphate and the inhibition of ribonucleic acid synthesis by proflavine. PMID- 11945926 TI - Dependence of actomyosin NTPase activity on ionic strength and its modification by alpha-actinin. PMID- 11945928 TI - Altered mitochondrial ribosomes in a cytoplasmic mutant of yeast. PMID- 11945929 TI - Quantum requirement of photosystem I mediated ATP formation in chloroplast fragments. PMID- 11945930 TI - Peptide cleavage in sheep erythrocyte membranes during the action of complement. PMID- 11945931 TI - A possible role for rhodanese: The formation of 'labile' sulfur from thiosulfate. PMID- 11945933 TI - Kinetical study on the reconstitution of methionine-acceptor activity from fragments of Escherichia coli tRNA(fMet) with a deletion in the dihydrouridine region or the amino acid-acceptor stem. PMID- 11945932 TI - On the chemical nature of alteration and modification of DNA dependent RNA polymerase of E. coli after T4 infection. PMID- 11945934 TI - Proteoglycans from sheep, pig, rat and human spleens having chemical and biological resemblances to that in Kurloff cells. PMID- 11945935 TI - Crystallisation of trypsin-modified methionyl-tRNA synthetase from Escherichia coli. PMID- 11945936 TI - Carbon monoxide-reactive haemoproteins in parasitic flagellate Crithidia oncopelti. PMID- 11945937 TI - Crystallisation of angiotensinamide II. PMID- 11945938 TI - Quantitative correlation of uptake with antibiotic activity of polymyxin B in Salmonella typhimurium. PMID- 11945939 TI - The amino acid sequence of bovine growth hormone. PMID- 11945941 TI - Evidence for polar reconstitution of TMV. PMID- 11945940 TI - Kinetic studies of ceruloplasmin-azide interaction. PMID- 11945942 TI - Liposome bilayer model systems of freezing living cells. PMID- 11945943 TI - Differential action of trypsin on the beta' subunit of DNA-dependent RNA polymerase from E. coli. PMID- 11945945 TI - MAK column chromatography of the first RNA synthesized during germination of Zea mays embryos. PMID- 11945944 TI - Effects of acrolein on transcription in vitro. PMID- 11945946 TI - Cell envelope proteins of dividing and non-dividing cells of Escherichia coli. PMID- 11945947 TI - Affinity chromatography of trypsin inhibitors on trypsinogen-agarose exchangers. PMID- 11945948 TI - Synthesis of the flavocoenzyme of monoamine oxidase. PMID- 11945949 TI - Late replication of the DNA associated with the nuclear membrane. PMID- 11945950 TI - Evidence of the existence of two forms of sucrose synthetase. PMID- 11945951 TI - An inhibitory effect of D-Mannoheptulose on lactate formation in human whole blood in vitro. PMID- 11945952 TI - Dolicholmonophosphates: Mannosyl acceptors in a particulate in vitro system of S. cerevisiae. PMID- 11945953 TI - Some structural requirements for the antibiotic action of distamycins. PMID- 11945954 TI - Opposite effects of bongkrekic acid and atractyloside on the adenine nucleotides induced mitochondrial volume changes and on the efflux of adenine nucleotides. PMID- 11945955 TI - The rate of deoxygenation of red blood cells: Effect of intracellular 2,3 diphosphoglycerate and pH. PMID- 11945956 TI - Proton magnetic resonance study of the histidine residues of horse myoglobin. PMID- 11945957 TI - The role of the tricarboxylate transporting system in the production of phosphoenolpyruvate by ox liver mitochondria. PMID- 11945958 TI - Dependence of actomyosin ATPase activity on ionic strength and its modification by thiol group substitution. PMID- 11945959 TI - Complementary trimer binding to transfer RNA(Phe). PMID- 11945960 TI - The conformation of alamethicin. PMID- 11945961 TI - A new method for partial digestion useful for sequence analysis of polynucleotides. PMID- 11945962 TI - The oxidation-reduction potential of the copper signal in pigeon heart mitochondria. PMID- 11945963 TI - Cleavage of glucagon by alpha- and beta-trypsin. PMID- 11945964 TI - Proteolytic activity of pseudotrypsin. PMID- 11945965 TI - On the relation between adenine nucleotide carrier sites and atractyloside binding in mitochondria. PMID- 11945966 TI - Interaction between the binding of 35S-atractyloside and bongkrekic acid at mitochondrial membranes. PMID- 11945967 TI - ? AB - DNSA (1-Dimethylamino-naphthelene-5-sulfonamide) forms with carbonic anhydrase (carbonate hydrolase, EC 4.2.1.1) a highly fluorescent complex [1]. We used the formation of these complexes for the detection of the isoenzyme bands of carbonic anhydrase in isoelectric focusing and electrophoresis in acrylamide gels. DNSA is added to the solution of the isoenzymes before separation. Using quartz tubes - the maximum of the excitation spectrum is 280 nm [1] - for supporting the gels the fluorescent enzyme bands can be observed during development in UV light and without putting the gels out of the quartz tubes. This technique allows the detection of the carbonic anhydrase isoenzymes with greater precision and simplicity than with any staining method. PMID- 11945968 TI - Tryptophanyl circular dichroism in a special hemoglobin. PMID- 11945970 TI - The regulation of nitrate reductase activity from spinach Spinacea oleracea L. leaves by thiol compounds in the presence of adenosine-5'-diphosphate. PMID- 11945969 TI - Homocitrulline formation following carbamylation of histones with carbamyl phosphate. PMID- 11945971 TI - Proton translocation and energy dependent quenching of chlorophyll a fluorescence. PMID- 11945972 TI - The subunits of an acetylcholinesterase preparation purified from trypsin-treated electric eel tissue. PMID- 11945974 TI - Inhibitory effects of alpha-amanitin on RNA synthesis and induction of DOPA decarboxylase by beta-ecdysone. PMID- 11945973 TI - Electrophoretic characterization of rapidly transported proteins in axons of retinal ganglion cells. PMID- 11945975 TI - The effect of arginine modification on the pH dependence of pepsin activity. PMID- 11945976 TI - Kinetics of 125I-insulin binding to rat liver plasma membranes. PMID- 11945977 TI - A colorimetric estimation of lipopolysaccharides. PMID- 11945978 TI - Glycosyltransferases of human plasma. PMID- 11945979 TI - Modification of catalytic properties of amine oxidases. PMID- 11945980 TI - Effect of calcium chelation by L-phenylalanine on (Na(+) + K(+))-stimulated ATPase of chick brain microsomes. PMID- 11945981 TI - Reaction mechanisms and control properties of phosphotransferases. A report of a Joint Biochemical Symposium of the All-Union Biochemical Society of the USSR and the Biochemical Society of the GDR, Schloss Reinhardsbrunn, GDR, May 3-7, 1971. PMID- 11945982 TI - Antigens and immunogenicity. Joint Biochemical Meeting of the Societe Belge de Biochimie - Belgische Vereniging voor Biochemie and the Gesellschaft fur Biologische Chemie, Liege, January 14-16, 1971. PMID- 11945983 TI - On the need for regulation of succinate dehydrogenase. PMID- 11945984 TI - The Ca(2+) stimulated incorporation of choline into microsomal lecithin subspecies in vitro. PMID- 11945985 TI - Structural studies on ovine growth hormone. Sequence of a cyanogen bromide fragment. PMID- 11945986 TI - Characterization of an amino-acid substitution leading to spectinomycin resistance in Escherichia coli. PMID- 11945988 TI - The formation of a long-lived complex between an ATP-dependent deoxyribonuclease and DNA. PMID- 11945987 TI - On the properties of trypsin inhibitors from human and bovine colostrum. PMID- 11945989 TI - Primary structure of N-terminal part of molecule of dolphin myoglobin. PMID- 11945990 TI - Preferential oxidation of fatty acids by rat small intestine. PMID- 11945992 TI - The lactate dehydrogenase in preimplantation mouse embryos of quackenbush and swiss mice. PMID- 11945991 TI - The primary sequence of phospholipase-A from bee venom. PMID- 11945993 TI - Conformation of derivatives of bovine alpha-lactalbumin obtained by cyanogen bromide attack. PMID- 11945994 TI - Energy-linked changes of the membrane of Rhodospirillum rubrum chromatophores detected by the fluorescent probe 8-anilinonaphthalene-1-sulfonic acid. PMID- 11945995 TI - Cholesterol esters and steroidogenesis in pig corpora lutea. PMID- 11945996 TI - Exchange between divalent metallic cations and sodium fixed on ionisable sites of DNA. PMID- 11945997 TI - Distinct cytoplasmic and nuclear DNA polymerases from rat liver. PMID- 11945998 TI - Adenosine-rich sequences in rapidly hybridizing messenger-like RNA and their possible significance for reiterated base sequences in eukaryotic DNA. PMID- 11945999 TI - Fluorescence studies of the binding of valyl-tRNA synthetase and tryptamine to valine-specific tRNA. A possible role for tryptophan residues in the binding of aminoacyl-tRNA synthetases to tRNAs. PMID- 11946000 TI - Preparation of pure bovine apo-erythrocuprein by gel filtration. PMID- 11946001 TI - Effect of amino acid starvation on constitutive synthesis of histidine mRNA. PMID- 11946002 TI - End-product inhibition of Pisum phenylalanine ammonia-lyase by the Pisum flavonoids. PMID- 11946003 TI - The primary structure of prochymosin (prorennin) EC 3.4.4.3. some tryptic fragments of a maleylated preparation. PMID- 11946004 TI - A simple procedure for the isolation of rat liver microsomes. PMID- 11946005 TI - Ribosome metabolism in mildew-infected barley leaves. PMID- 11946006 TI - The occurrence of circular structures in highly reiterated DNA of the rat. PMID- 11946007 TI - A glycoprotein located in the intermembrane space of rat liver mitochondria. PMID- 11946008 TI - Muscle fibrils: Solubilization and gel electrophoresis. PMID- 11946009 TI - Chain elongation factors of yeast mitochondria. PMID- 11946011 TI - Biosynthesis of plasmalogens from alkyl- and alkyl-acyl-glycerophosphoryl ethanolamine in the rat brain. PMID- 11946010 TI - Biosynthesis of sideramines in fungi. Mevalonate as a precursor of cis- and trans 5-hydroxy-3-methyl-2-pentenoic acids. PMID- 11946012 TI - Fluorescence of lucensomycin upon binding to erythrocyte ghosts. PMID- 11946013 TI - Cytochrome biosynthesis under copper-limited conditions in Candida utilis. PMID- 11946015 TI - Glycine and serine inhibition of D-glycerate dehydrogenase and 3-phosphoglycerate dehydrogenase of rat brain. PMID- 11946014 TI - Cytoplasmic aspartate aminotransferase from pig heart muscle: Partial sequence. PMID- 11946016 TI - Induction of rat liver tyrosine aminotransferase by dibutyryl cyclic AMP and its inhibition by actinomycin D and alpha-Amanitin. PMID- 11946017 TI - A study of the secondary structure of ilamycin B(1) by 300 MHz proton magnetic resonance. PMID- 11946018 TI - Natural abundance 13C spectra of proteins: Carboxy-myoglobin and hemoglobin. PMID- 11946019 TI - Escherichia coli tyrosyl-tRNA synthetase. Influence of magnesium ions on the enzymatic activity. PMID- 11946020 TI - pH-dependence of aerobic glycolysis in ehrlich ascites tumour cells. PMID- 11946021 TI - Eukaryotic ribosomal proteins. Two-dimensional electrophoretic studies. PMID- 11946022 TI - Pressure-induced dissociation of ribosomes during ultracentrifugation. PMID- 11946023 TI - Interaction of haemoglobin with ions binding of inositol hexaphosphate to human haemoglobin a. PMID- 11946024 TI - The effects of tetrazolium salts on monoamine oxidase activity. PMID- 11946025 TI - Substrate specificity of plant peroxidases. PMID- 11946026 TI - Crystallized acidic nucleoprotein from pig eye lenses. PMID- 11946027 TI - The detachment of ribosomal proteins by urea: evidence for non-electrostatic RNA protein interaction in the ribosome. PMID- 11946028 TI - A cytoplasmic molecule active on membranar Mg(2+) movements. I - isolation and properties. PMID- 11946029 TI - A cytoplasmic molecule active on membranar Mg(2+) movements. II - its role as a factor of the membrane integrity in mitochondria, chloroplasts and bacteria. PMID- 11946030 TI - Role of Ca(2+) in the oxidation of exogenous NADH by plant mitochondria. PMID- 11946031 TI - Biocide resistance in yeast: Isolation and general properties of trialkyl tin resistant mutants. PMID- 11946032 TI - Further evidence for the four-site model of ribosomal function: Bacteriophage fr mRNA coded binding of AA-tRNA. PMID- 11946033 TI - Recognition by T factor of a tRNA(phe)(yeast) molecule recombined from 3'and 5' halves; and its non messenger-dependent binding to ribosomes. PMID- 11946034 TI - Specific protection against inhibitors of the NADH-nitrate reductase complex from spinach. PMID- 11946035 TI - Spectral properties of the coenzyme bound to DOPA decarboxylase from pig kidney. PMID- 11946036 TI - Introduction of an intramolecular flourescent pobe in E. coli tRNA(Val)(1). PMID- 11946037 TI - Polarographic activity and electrolytic reduction of ferredoxin. PMID- 11946038 TI - Further evidence for a transport form of collagen. Its extrusion and extracellular conversion to tropocollagen in embryonic tendon. PMID- 11946039 TI - Reduced levels of 5,6-dihydrouridine in fluorouracil-containing transfer RNAs from Saccharomyces cerevisiae. PMID- 11946040 TI - The sideness of mitochondrial membranes and their sub-particles as characterized by preparative free flow electrophoresis. PMID- 11946041 TI - Sulphydryl groups and the in vitro enzymatic synthesis of 5-amino-laevulinic acid and porphobilinogen in Rhodopseudomonas spheroides. PMID- 11946042 TI - Synaptosomal proteins and axoplasmic flow: Fractionation by SDS polyacrylamide gel electrophoresis. PMID- 11946043 TI - Nucleotide sequence of "renaturable" leucine transfer ribonucleic acid. PMID- 11946045 TI - Pyrrolooxygenases: The biosynthesis of 2-aminoacetophenone. PMID- 11946044 TI - On the heterogenity of individual tRNA(val)(1) that arises in the course of modification with CME-carbodiimide. PMID- 11946046 TI - Ribosomal DNA satellite of Euglena gracilis chloroplast DNA. PMID- 11946047 TI - Gummiferin, an inhibitor of the adenine-nucleotide translocation. Study of its binding properties to mitochondria. PMID- 11946048 TI - The difference in buoyant density between viral and normal polyribosomes in krebs II cells. PMID- 11946049 TI - The effect of ionic strength on the buoyant density of polyribosomes and 80 s free ribosomes in EMC virus-infected and uninfected krebs II cells. PMID- 11946050 TI - The production of superoxide radicals in reactions of the herbicide diquat. PMID- 11946051 TI - Intramitochondrial localization of phosvitin kinase and phosvitin phosphatase. PMID- 11946052 TI - Regulation of hepatic elimination of ethanol in vivo. PMID- 11946053 TI - The nucleotide sequences of two tryptophane-tRNAs from brewer's yeast. PMID- 11946054 TI - Stoichiometric studies on aryl-aldehyde: NADP oxidoreductase from Neurospra crassa. PMID- 11946055 TI - Transport of nuclear DNA into the cytoplasm in cultured human liver cells. PMID- 11946056 TI - Reaction of aspartate aminotransferase apoisozymes with glyceraldehyde-3-P: Effect of NaBH(4) treatment and subsequent reactivation by pyridoxal-5'-P. PMID- 11946057 TI - Hyaluronidase chemically bound to agarose. PMID- 11946058 TI - A new DNA intercalating drug: Methoxy-9-ellipticine. PMID- 11946060 TI - The subunit composition of the mitochondrial oligomycin-insensitive ATPase. PMID- 11946059 TI - Stimulation of "non-enzymic" translocation in ribosomes by p chloromercuribenzoate. PMID- 11946061 TI - Reassessment of the molecular weight of mitochondrial ATPase from beef heart. PMID- 11946062 TI - Tritium labelling of the alpha-neurotoxin of Naja nigricollis. PMID- 11946063 TI - Tautomerism of nucleic bases and the template synthesis of polynucleotides. PMID- 11946064 TI - Activation of a differentiation-inducing protein by adenine and adenine containing nucleotides. PMID- 11946065 TI - The effect of arsenite on rat-liver mitochondria. PMID- 11946066 TI - Long chain acyl CoAs, adenine nucleotide translocase and the coordination of the redox states of the cytosolic and mitochondrial compartments. PMID- 11946067 TI - High pressure dissociation of lactic dehydrogenase. PMID- 11946068 TI - Modes of action of erythromycin and thiostrepton as inhibitors of protein synthesis. PMID- 11946069 TI - Tosylphenylalanyl chloromethane-inhibitor of complex of S(1)S(3)-factors in cell free protein-synthetizing system from Bacillus stearothermophilus. PMID- 11946071 TI - RNA-binding protein factor of animal cell extracts. PMID- 11946070 TI - A new form of pyruvate kinase in mycelium of Mucor rouxii. PMID- 11946072 TI - Reversible cyanide inhibition of spinach (Spinacea oleracea L.) nitrate reductase and non-exchangeability in vitro of protein bound molybdenum and tungsten. PMID- 11946073 TI - The polypeptide chain molecular weight of a mammalian hexokinase. PMID- 11946074 TI - ? AB - The interaction of caffeine with DNA has been studied at several ionic strengths by heat denaturation. At 2 mM K(+) caffeine produces a stabilization, but at 20 mM K(+) or higher a destabilization of secondary structure of DNA occurs. There is a correlation between heterogeneity and destabilization of DNA in presence of caffeine. In the premelting region a hyperchromic effect can be observed. It is suggested that two binding processes exists in the DNA-caffeine interaction. PMID- 11946075 TI - Affinity gel filtration: A new method for the rapid determination of apparent molecular weights of enzymes. PMID- 11946076 TI - Inhibition of citrate synthase by succinyl-CoA and other metabolites. PMID- 11946077 TI - The protein moiety of nuclear ribonucleoprotein particles containing DNA-like RNA: Presence of heterogeneous and high molecular-weight polypeptide chains. PMID- 11946078 TI - Identification of the protein moiety of the LP(a)-lipoprotein in human plasma. PMID- 11946079 TI - ? PMID- 11946080 TI - Localization of a microheterogeneity in the amino acid sequence of bovine growth hormone. PMID- 11946081 TI - On the pathway of electron transport in cyclic photophosphorylation. PMID- 11946082 TI - Amino acid sequence homology between muscle and liver aldolases. PMID- 11946083 TI - Studies on polypeptide chain initiation factors F(1) and F(2). PMID- 11946084 TI - Mannolipid synthesis in a cell-free system of Mycobacterium smegmatis. PMID- 11946085 TI - AMP Like contamination in commercial NAD preparations. PMID- 11946087 TI - Epoxides as microsomal metabolites of polycyclic hydrocarbons. PMID- 11946086 TI - The nature of the breakdown of thyroglobulin. PMID- 11946088 TI - Prevention of Ca(2+) induced cation efflux from liver mitochondria by a cytoplasmic factor and by oligomycin. PMID- 11946089 TI - Proteins associated with globin messenger RNA in avian erythroblasts: Isolation and comparison with the proteins bound to nuclear messenger-likie RNA. PMID- 11946090 TI - Insulin release from pieces of pancreas incubated in vitro in the presence of D erythrose. PMID- 11946091 TI - Enzymic synthesis of ethanolamine plasmalogens from an O-alkyl glycerolipid. PMID- 11946092 TI - Position of glycoprotein polypeptide chain in the human erythrocyte membrane. PMID- 11946093 TI - Kinetic evidence for Ca(2+) uptake by intact Ehrlich ascites tumor cells. PMID- 11946094 TI - The degradation of alpha-pinene by Pseudomonas PX1. PMID- 11946095 TI - ? AB - Cytochrome c of the zebra (Equus guagga Boehmi) differs from horse cytochrome c in having in its polypeptide chain one serine residue instead of none and 9 threonine residues instead of 10. This replacement has been localised at position 47 of the sequence. PMID- 11946096 TI - Effect of cholesteryl 14-methylhexadecanoate on the RNA polymerase activity of rat liver nuclei in vivo and in vitro. PMID- 11946098 TI - The effect of 2,4-dichlorophenoxyacetic acid on the basicity of proteins during callus induction. PMID- 11946097 TI - Separation of purified human chorionic gonadotropin into single bands by isoelectric focusing and their characterization. PMID- 11946099 TI - Use of 2-[(4-aminophenyl)-sulfonyl]-ethyl hydrogen sulfate for the preparation of a dextran-specific immunogen. PMID- 11946100 TI - Effects of ethanol on mouse liver polysomal disaggregation by dimethylnitrosamine and lasiocarpine. PMID- 11946101 TI - Effect of glucagon on phosphorylation of some rat liver ribosomal proteins in vivo. PMID- 11946102 TI - A Thermosensitive mutant defective in ribosomal 30 S subunit assembly. PMID- 11946103 TI - Chick embryo poly (rA:dT)-dependent DNA polymerase. PMID- 11946104 TI - Messenger RNA from arginine and phosphoenolpyruvate carboxylase genes in arg R+ and arg R(-) strains of E. coli K-12. PMID- 11946105 TI - The molecular weights and association of the histones of chicken erythrocytes. PMID- 11946106 TI - Studies on the bacteriophage MS2 nucleotide sequence of a 3'-terminal fragment (n = 104). PMID- 11946107 TI - Formation of a complex between 23 S RNA, 5 S RNA and proteins from Escherichia coli 50 S ribosomal subunits. PMID- 11946108 TI - Anion translocators in rat-heart mitochondria. PMID- 11946109 TI - Comparative studies on the nature of the cross-links stabilizing the collagen fibres of invertebrates, cyclostomes and elasmobranchs. PMID- 11946110 TI - Synthesis of lens protein in vitro V. Isolation of messenger-like RNA from lens by high resolution zonal centrifugation. PMID- 11946111 TI - Comparison of peptide patterns from isolated 30 S and 50 S ribosomal proteins of Escherichia coli by column chromatography. PMID- 11946112 TI - On the mechanism of the inhibiting effect of aurin tricarboxylic acid on the binding of ribosomes to ribonucleic acids. PMID- 11946113 TI - Interference of ethidium bromide with the formation of supercoiled mitochondrial DNA. PMID- 11946114 TI - Properties of a 2,3-oxidosqualene-cycloartenol cyclase from Ochromonas malhamensis. PMID- 11946115 TI - The biochemistry and biophysics of mitochondrial membranes. A report on the International Symposium held in Bressanone, Italy, June 1971. PMID- 11946116 TI - Enhancement of ribosome-dependent GTPase from E. coli by streptomycin. PMID- 11946117 TI - Reptilian hemoglobins: N-terminal sequence of an alpha-chain of viper (Vipera aspis) hemoglobin. PMID- 11946118 TI - Structural modifications involved in the dissociation and reassociation of the alpha and beta subunits of ovine luteinizing hormone. PMID- 11946119 TI - Interactions between aminoacyl tRNA synthetases, tRNAs, and fluorescent dyes. PMID- 11946121 TI - DNA-dependent RNA polymerases from yeast. Partial characterization of three nuclear enzyme activities. PMID- 11946120 TI - Fluorescence of proteins interacting with nucleic acids. Correction for light absorption. PMID- 11946122 TI - Chemical evidence for syncatalytic conformational changes in aspartate aminotransferase. PMID- 11946123 TI - Replacement of Y base, dihydrouracil, and 7-methylguanine in tRNA by artificial odd bases. PMID- 11946125 TI - Effects of chlorosuccinate in the kidney cortex. PMID- 11946124 TI - Tritium as a tracer for mitochondrial and cytosolic reducing hydrogen in the kidney cortex. PMID- 11946126 TI - Electronoptical studies of procollagen from the skin of dermatosparaxic calves. PMID- 11946127 TI - A survey of catechol ring-cleavage by sterile plant tissue cultures. PMID- 11946128 TI - DNA polymerase activity from Tetrahymena pyriformis. PMID- 11946129 TI - ATP requirement in the course of calcium uptake by human myometrial mitochondria. PMID- 11946130 TI - Human liver carbamyl phosphate-glucose phosphotransferase activity: Catalytic properties and physiological phosphorylative potential. PMID- 11946131 TI - Electrophoretic separation of myosin large subunits. PMID- 11946132 TI - Studies on the mode of interaction of daunomycin with DNA. PMID- 11946133 TI - Covalent fixation of pepsin to agarose derivatives. PMID- 11946134 TI - On the mechanism of action of vitamin K in vertebrates and bacteria. PMID- 11946135 TI - Respiratory chain linked H(2)O(2) production in pigeon heart mitochondria. PMID- 11946136 TI - Tryptic cleavage of homoarginyl bonds in the oxidized guanidinated derivative of a bovine trypsin inhibitor (Kunitz inhibitor). PMID- 11946137 TI - The transition metal-binding site of concanavalin A at 2.8 A resolution. PMID- 11946139 TI - The utilization of L-adenosine by mammalian tissues. PMID- 11946138 TI - Coordinate regulation of DNA-dependent cell-free synthesis of uridyltransferase and galactokinase. PMID- 11946140 TI - The regulatory role of amylo-1,6-glucosidase/oligo-1,4-->1,4-glucantransferase in liver glycogen metabolism. PMID- 11946141 TI - Oligoribonucleotides as primer for terminal deoxynucleotidyl transferase. PMID- 11946143 TI - Degree of methylation of mitochondrial ribosomal RNA. PMID- 11946142 TI - The heterogeneity of erythrocyte IMP: pyrophosphate phosphoribosyl transferase and purine nucleoside phosphorylase by isoelectric focusing. PMID- 11946144 TI - Identification of non-methylene-interrupted cis,cis-octadecadienoic acids in human milk. PMID- 11946145 TI - The mode of action of cyclic amp in the rat anterior pituitary. PMID- 11946146 TI - Non-oxidative demethylation of trimethylamine N-oxide by Pseudomonas aminovorans. PMID- 11946147 TI - The complexity of human mitochondrial DNA. PMID- 11946148 TI - Effect of disulfide cleavage on the molecular weight of one of the major polypeptides of human serum high density lipoprotein. PMID- 11946149 TI - The effects of ACTH and of cortisol on serum ceruloplasmin in rabbits. PMID- 11946150 TI - Mercury-labelled concanavalin A as a marker in electron microscopy-localisation of mannan in yeast cell walls. PMID- 11946151 TI - Commercial bilirubin: A trinity of isomers. PMID- 11946153 TI - Evidence for proteolytic fragments in commercial samples of human ceruloplasmin. PMID- 11946154 TI - Complex and cluster formation in mixed lecithincholesterol bilayers. Cooperativity of motion in lipid systems. PMID- 11946155 TI - The use of NMR spectra of sonicated phospholipid dispersions in studies of interactions with the bilayer. PMID- 11946152 TI - The proteolytic action of the snake venom enzymes arvin and reptilase on N terminal chain-fragments of human fibrinogen. PMID- 11946156 TI - Some regulatory properties of purine nucleoside phosphorylase of Bacillus cereus. PMID- 11946157 TI - The effect of alpha-amanitin on RNA synthesis in rat liver mitochondria. PMID- 11946159 TI - The amino acid sequence of cytochrome c(3) from Desulfovibrio desulfuricans (strain el agheila Z, NCIB 8380). PMID- 11946158 TI - Comparative studies on the isoleucyl-tRNA synthetases from Bacillus stearothermophilus and Escherichia coli: II. The effect of magnesium ions in the transacylation reaction. PMID- 11946160 TI - The amino acid sequence of cytochrome c-551.5 (Cytochrome c(7)) from the green photosynthetic bacterium Chloropseudomonas ethylica. PMID- 11946161 TI - Regulation of pyruvate kinase during glyconeogenesis in Saccharomyces cerevisiae. PMID- 11946162 TI - The effect of temperature on energy-linked functions in chloroplasts. PMID- 11946163 TI - ? AB - Occurence of PAL, cinnamate-hydroxylase and p-coumarate-hydroxylase, is found in cell-free extracts from Quercus pedunculata roots; moreover, an enzyme system which catalyzes benzoic acid formation from cinnamic acid is caracterized for the first time. Role of these enzymes and their interactions within the same organ are discussed. PMID- 11946164 TI - Polydeoxythymidylate as template for complementary enzymatic synthesis. PMID- 11946166 TI - Identification of a third form of glycogen synthetase in rat chloroma tumors. PMID- 11946165 TI - A thermodynamic comparison of some oxidations of ferrocytochrome C. PMID- 11946167 TI - Mitochondrial autonomy: Synthesis of DNA from RNA templates in isolated mammalian mitochondria. PMID- 11946168 TI - The determination of 5-azapyrimidines and their derivatives in bacterial RNA. PMID- 11946170 TI - Infectivity to Escherichia coli spheroplasts of linear phiX174 DNA strands derived from the replicative form (RFII) of phiX DNA. PMID- 11946169 TI - Synthesis of lens protein in vitro VI. Methionyl-tRNA from eye lens. PMID- 11946171 TI - ? AB - Candida tropicalis strain 101 possesses a hydroxylase system when grown on tetradecane as the carbon source which is active towards hydrocarbons and fatty acids. This system including cytochrome P(450) and NADPH-cytochrome c reductase has been localized exclusively in the microsomal fraction. PMID- 11946172 TI - Partial digestion of 32P-fd DNA with T(4) endonuclease IV. PMID- 11946174 TI - Betulaprenol phosphate as an acceptor of mannose from GDP-mannose in Phaseolus aureus preparations. PMID- 11946173 TI - Nitroxides XLVIII: A study of the interaction between bovine serum albumin and a modified steroid by electron spin resonance. PMID- 11946175 TI - Trihydroxy long-chain bases in bovine milk sphingomyelin. PMID- 11946177 TI - Subcellular localization of cystathionine synthase in rat brain. PMID- 11946176 TI - Synthesis and turnover of liver chromatin acidic proteins. PMID- 11946178 TI - ? AB - It is shown that the iso-1 and iso-2 cytochromes c synthesized by two strains of respiratory deficient strains of yeast (varrho(-)) possess one residue of epsilonN-trimethyllysine and do not differ in this respect from the iso cytochromes produced by varrho(+) strains. This result excludes the possibility that the methylation reaction is linked to respiratory activity or to the varrho(+) factor. PMID- 11946179 TI - Reconstitution of the 50 S ribosome subunit. Localization of activities related to the peptidyl transferase centre. PMID- 11946180 TI - Reconstitution of the 50 S ribosome subunit. Localization of G-dependent GTPase activity. PMID- 11946183 TI - Erratum. PMID- 11946182 TI - Changes in transcription during early embryogenesis of sea urchins. PMID- 11946181 TI - Characterization of a cytoplasmic histone-coenzyme A acetyltransferase. PMID- 11946184 TI - PAPS-cerebroside sulphotransferase activity in brain and kidney of neurological mutants. PMID- 11946185 TI - The oxygen equilibrium of yeast hemoglobin. PMID- 11946186 TI - The isolation of informofers free from dRNA. PMID- 11946187 TI - Effect of loss of cytochrome c following storage of mitochondria in situ. PMID- 11946188 TI - ? PMID- 11946189 TI - Large scale isolation of the caulobacter bacteriophage OCb5 and its RNA genome. PMID- 11946190 TI - Association products of native and derived ribosomal subunits of E. coli and their stability during centrifugation. PMID- 11946192 TI - The relationship between C-550 and delayed fluorescence. PMID- 11946191 TI - An approach to specific labelling of ribosome in the region of peptidyl transferase center using N-acylaminoacyl-tRNA with an active alkylating grouping. PMID- 11946193 TI - Effect of muscular exercise and glycogen depletion on rat liver and kidney phosphoenolpyruvate carboxykinase. PMID- 11946194 TI - First demonstration of UDP-GAL: sn-glycero-3-phosphoric acid 1-alpha-galactosyl transferase and its possible role in osmoregulation. PMID- 11946195 TI - L-ribonucleosides do not penetrate bacterial cell walls. PMID- 11946196 TI - Effects of synthetic analogues of phosphoenolpyruvate on muscle and liver pyruvate kinase, muscle enolase, liver phosphoenolpyruvate carboxykinase and on the intra-/extra-mitochondrial tricarboxylic acid carrier transport system. PMID- 11946197 TI - Cytokinins that inhibit transfer RNA methylating enzymes. PMID- 11946198 TI - Circular dichroism studies on the complex between beef liver glutamate dehydrogenase and NADH. PMID- 11946199 TI - A comparative study of the protein components of nuclear and polysomal messenger ribonucleoprotein. PMID- 11946200 TI - Inhibition by saccharides of the agglutination of periodate-treated erythrocytes by horse serum. PMID- 11946201 TI - In vitro synthesis of T3 AND T7 RNA polymerase at low magnesium concentration. PMID- 11946202 TI - Isolation of ribose-poor rough microsomes from rat liver. PMID- 11946203 TI - Crystallization of L-arginine deiminase from Pseudomonas Putida. PMID- 11946204 TI - The major proteins of human and monkey amyloid substance: Common properties including unusual N-terminal amino acid sequences. PMID- 11946206 TI - Erratum. PMID- 11946205 TI - Selective inhibition of the reactions catalyzed by ribosome-specific transfer factors G. PMID- 11946207 TI - Molecular genetics of antibody formation Report of a Colloquium held during the 7th FEBS Meeting, Varna, Bulgaria, 20-25 September 1971. PMID- 11946208 TI - Transitions in aliphatic and polar head regions of phosphatidyl choline vesicles determined with fluorescent probes. PMID- 11946209 TI - Oxidative phosphorylation in intact Azotobacter vinelandii. PMID- 11946210 TI - Different classes of initiation factors F3 and their dissociation activity. PMID- 11946211 TI - Studies on the ribosomal initiation factor F(3). Multiplicity of F(3) and DF activities. PMID- 11946212 TI - Internal homology in the alpha chain of bovine luteinizing hormone. PMID- 11946213 TI - The amino acid sequence of the bovine luteinizing hormone beta subunit. PMID- 11946214 TI - Distribution of ammonia and methylamine between mitochondria and suspension medium. PMID- 11946215 TI - Biosynthesis of glucuronosyl diglyceride by a cell-free system obtained from a moderately halophilic-halotolerant bacterium. PMID- 11946216 TI - Some aspects of mitochondrial structure. PMID- 11946217 TI - Isolation and properties of Escherichia coli mutants defective in 2-keto 3-deoxy 6-phosphogluconate aldolase activity. PMID- 11946218 TI - Isolated adrenal cells: Log dose response curves for steroidogenesis induced by ACTH(1-24), ACTH(1-10), ACTH(4-10) and ACTH(5-10). PMID- 11946219 TI - Isolated adrenal cells: ACTH(11-24), a competitive antagonist of ACTH(1-39) and ACTH(1-10). PMID- 11946220 TI - A new catalytic activity of alpha-glycerolphosphate dehydrogenase: The enzymatic hydrolysis of p-nitrophenyl acetate. PMID- 11946221 TI - Regulation of succinate oxidation by endogenous reduced nicotinamide adenine dinucleotides in intact heart mitochondria. PMID- 11946222 TI - Specific inhibition of poly adpribose polymerase by thymidine and nicotinamide in HeLa cells. PMID- 11946223 TI - ? AB - We have applied successfully the active enzyme analytical centrifugation method (A.E.C.) to the study of the well known pyruvate dehydrogenase complex (P.D.C.) in crude extracts from E. coli. It is not possible to study the complex in a spectrophotometer (through its dehydrogenase activity) due to NADH oxidase activities. The A.E.C. method, which overcomes this difficulty, can also be viewed as a refined spectrophotometric method. PMID- 11946224 TI - "Malic enzyme" and alkaline phosphatase in mitochondria from pig and rat ovaries. PMID- 11946225 TI - Introduction of elastolytic activity into alkaline proteinase from Aspergillus sojae via electrostatic changes due to modification of carboxyl groups. PMID- 11946226 TI - The effect of proteolytic enzymes on the troponin complex. PMID- 11946227 TI - 31P magnetic resonance of purified tRNA. PMID- 11946228 TI - Genetic control of pigment-protein complexes I and Ia of the plastid mutant en:alba-1 of Antirrhinum majus. PMID- 11946229 TI - Preparation and properties of nicotinamide guanine dinucleotide. PMID- 11946230 TI - Chlorophyll B fluorescence and an emission band at 700 nm at room temperature in green algae. PMID- 11946231 TI - Purification of human erythrocyte phosphofructokinase. PMID- 11946232 TI - Evidence for different oligomeric forms of human erythrocyte phosphofructokinase. PMID- 11946233 TI - Increase of the rotational relaxation time of antibody molecule after complex formation with dansyl-hapten. PMID- 11946234 TI - Multiple states in macromolecules I. Qualitative model for a single nucleation process. PMID- 11946235 TI - Multiple states in macromolecules II. Entropic behaviour of tRNA degraded by polynucleotide phosphorylase. PMID- 11946236 TI - The concentration dependence of the activity of beef liver glutamate dehydrogenase as measured by rapid mixing and ultracentrifugation techniques. PMID- 11946237 TI - Human tetanus antibodies: Isolation and characterization with special reference to the IgG subclasses. PMID- 11946238 TI - Estimation of dissociation constant of enzyme-ligand complex from fluorometric data by "difference" method. PMID- 11946239 TI - Inhibition of amino acid transport in Escherichia coli cells and its cell membranes. PMID- 11946240 TI - Restoration by silico tungstate of the reversible light induced pH rise in coupling factor (CF(1)) deficient chloroplasts. PMID- 11946241 TI - Mitochondrial effects of copper-limited growth of Torulopsis utilis. PMID- 11946242 TI - Studies of the composition and structure of plasma lipoproteins. C- and N terminal amino acids of C-I polypeptide ("R-Val") of human plasma apolipoprotein C. PMID- 11946243 TI - Some structural requirements for the antibiotic action of distamycins II. Structural modification of the side chains in distamycin A molecule. PMID- 11946244 TI - Adenosine, a glucogenic and lipogenic compound. PMID- 11946245 TI - Regulation of arginase levels by urea and intermediates of the Krebs-Henseleit cycle in Saccharomyces cerevisiae. PMID- 11946246 TI - gamma-Glutamyltransferase in kidney brush border membranes. PMID- 11946247 TI - Process monitoring in solid phase peptide synthesis, amino group blocking effect of impure methylene chloride. PMID- 11946248 TI - On latent hexokinase activity in skeletal muscle mitochondria. PMID- 11946250 TI - Errata. PMID- 11946249 TI - Unusual CCA-stem structure of E. coli B tRNAH(His)(1). PMID- 11946251 TI - Isolation of liver and muscle polyribosomes in high yield after cell disruption by nitrogen cavitation. PMID- 11946252 TI - Sulfatides and sodium ion transport, sphingolipid composition of the rectal gland of spiny dogfish. PMID- 11946253 TI - Contribution to the study of the structure of adjuvant active waxes D from mycobacteria: Isolation of a peptidoglycan. PMID- 11946254 TI - Analysis of nucleic acid derivatives at the subnanomolar level. (IV) Analysis of polyribonucleotides by conversion to tritiated nucleoside derivatives. PMID- 11946255 TI - Stripping of ribosomal proteins from Escherichia coli ribosomes. PMID- 11946257 TI - ? PMID- 11946256 TI - ? AB - The blockage of the carboxyl groups of a pancreatic trypsin inhibitor (inhibitor of Kunitz) has been performed with a water-soluble carbodiimide and glycine ethyl ester. The inhibitor has five carboxyl groups located on residues Asp(3), Glu(7), Glu(49), Asp(50) and Ala(58). When the reaction is carried out on native inhibitor or on inhibitor treated with 8 M urea, approximately 3 moles of glycine ethyl ester are introduced but 4, 3 residues can be incorporated into performic acid oxidized inhibitor. Fully active derivatives with a complete substitution of the C-terminal residue can be obtained. It may be concluded that the C-terminal carboxyl group is not involved in the binding with trypsin. PMID- 11946258 TI - Two new metabolites of Aspergillus flavus (Link). PMID- 11946259 TI - Distribution of pyruvate carboxylase and phosphoenol-pyruvate carboxikinase in human liver. PMID- 11946260 TI - alpha-Ketoglutarate in biological hydroxylations. PMID- 11946261 TI - Amino acid sequence at the N-terminal end of a cold agglutinin Kappa chain. PMID- 11946262 TI - Evidence for induction of the 2,3-butanediol-forming enzymes in Aerobacter aerogenes. PMID- 11946263 TI - Amino acid acceptor activity of bacteriophage T4 transfer RNA. PMID- 11946264 TI - Lipotropic hormone obtained from human pituitary gland. PMID- 11946266 TI - Aminoacyl-tRNA-dependent interaction of a transfer enzyme from yeast with a complex formed by two other transfer factors with guanosine triphosphate. PMID- 11946265 TI - The effect of limited nitration on antibodies to dinitrophenyl group. PMID- 11946267 TI - ? AB - After synthesis of short, nascent oligonucleotide in the presence of (32P)DNA, GTP, CTP, UTP and 3'dATP, one can excise with deoxyribonuclease a ternary complex of RNA polymerase, protected DNA and oligonucleotide, while the enzyme simply bound to the template is removed by increasing the ionic strength. This ternary complex is retained on nitrocellulose membranes. On polyacrylamide gel electrophoresis it migrates faster than RNA polymerase alone. The protected portion of the DNA is constituted of about 75 nucleotides. It might represent the sites for RNA initiation. PMID- 11946268 TI - Cyclic AMP as an antagonist of catabolite repression in Escherichia coli. PMID- 11946269 TI - The number and nature of alpha,beta-unsaturated amino acids in nisin. PMID- 11946270 TI - Time dependence of activation of muscle AMP-aminohydrolase by substrate and potassium ion. AB - The kinetics of AMP-aminohydrolase, which under steady state conditions shows a typical sigmoid dependence of initial velocities versus substrate concentration, have been examined by rapid mixing methods. Using this technique it was observed that when substrate or substrate plus activator (K(+)) were mixed with enzyme, the rate of appearance of product markedly increased during the first few tenths of a second. The time course of this change in rate was taken to reflect the progress of activation by substrate or by K(+). On the other hand, addition of activator to enzyme prior to mixing with substrate gave process curves for the formation of product consistent with normal Michaelis-Menten behaviour.Under the conditions where the reaction was examined, the enzyme at time zero had less than 10% of the activity of the fully active enzyme. The time course for activation with K(+) followed a first order process with a rate constant of 10.6 sec(-1) at 20 degrees C. A simple mechanism consistent with the data and capable of explaining the sigmoid dependence of initial velocities versus substrate concentrations observed in steady state kinetics was proposed. PMID- 11946271 TI - The kinetic analysis of hydrolytic enzyme catalyses: Consequences of non productive binding. PMID- 11946272 TI - Gel chromatographic separation of oligosaccharides at elevated temperature. PMID- 11946273 TI - Specific binding of acetylcholine to acetylcholinesterase in the presence of eserine. PMID- 11946274 TI - Effect of the concentration of D-glyceraldehyde-3-phosphate dehydrogenase on the reducibility of its firmly bound NAD. PMID- 11946275 TI - The regulation of rat-liver xanthine oxidase: Activation by proteolytic enzymes. PMID- 11946276 TI - Uridine 2',3'-O,O-cyclophosphorothioate as substrate for pancreatic ribonuclease (I). PMID- 11946277 TI - Dedifferentiation of phospholipid composition in subcellular particles of cancer cells. PMID- 11946278 TI - Incorporation of [5-(3)H] uridine into DNA. PMID- 11946279 TI - N- and C-terminal sequences in pig immunoglobulin pi-chains: Assignment to the K type. PMID- 11946280 TI - Mechanism of hormone actions (III) The changes in the concentrations of free amino acids in rat liver and muscle and rates of autolysis of muscle protein after fasting and hormone treatment. PMID- 11946281 TI - Inhibition of arylaminopeptidases and proline iminopeptidases of human whole saliva by benzethonium chloride (benzyldimethyl [2-[2-(p-1,1,3,3 tetramethylbutylphenoxy)ethoxy]-ethyl] ammonium chloride). PMID- 11946282 TI - Origin of mitochondrial enzymes (II). the subcellular distribution of cytochrome c in rat liver tissue. PMID- 11946283 TI - Phosphofructokinase from E. Coli: Evidence for a tetrameric structure of the enzyme. PMID- 11946284 TI - A method for crystallization of serine-transfer-RNA. Co-crystallization of t-RNA with cadmium- and copper ion in water-dioxane. PMID- 11946285 TI - Further microheterogeneity of cytoplasmic aspartate aminotransferase from pig heart. PMID- 11946286 TI - Incorporation of 32P-phosphate into phosphatides of rat liver mitochondria in vivo and in vitro. PMID- 11946287 TI - The effect of ethanol on the concentrations of adenine nucleotides in rat liver. PMID- 11946288 TI - Photoreactivation spectrum of the co-inhibited taurochenodeoxycholate 6beta hydroxylase system. PMID- 11946289 TI - Effects of inducer substances on porphyrin synthesis in liver cell cultures. PMID- 11946290 TI - ? AB - Bovine insulin can be split into several fractions by gel filtration on Sephadex G50. These are, in order of decreasing molecular weight, a new unidentified substance, proinsulin, and a large fraction containing several substances. Ultracentrifugal examination of the proinsulin in acetic acid solution (pH 2.3) gave a molecular weight of 9300 and evidence of an association-dissociation equilibrium. PMID- 11946291 TI - The effects of ribosomes on the activity of a membrane bound enzyme catalysing thiol-disulphide interchange. PMID- 11946293 TI - Errata. PMID- 11946292 TI - ? AB - The terminal cis-glycol-group of phenylalanine specific tRNA (tRNA(Phe), I) from yeast was oxidized to the dialdehyde by NaIO(4) (tRNA(Phe)(oxi), II) and subsequently reduced to the diol by NaBH(4) (tRNA(Phe)(oxi-red), III). In charging experiments it could be shown that tRNA(Phe)(oxi-red) is still completely specific for phenylalanine. The Michaelis constant of tRNA(Phe)(oxi red) remains unchanged when compared with tRNA(Phe), while the maximun velocity of charging upsilon(max) of tRNA(Phe)(oxi-red) is decreased to about one half. PMID- 11946294 TI - Heterogeneity of 32PO(3-)(4) incorporation in alphaPO(3-)(4) of free rat liver nucleotides. PMID- 11946295 TI - Kinetics of inhibition of papain by TLCK and TPCK in the presence of BAEE as substrate. PMID- 11946296 TI - The transport of retinol in human plasma. PMID- 11946297 TI - The action of phenylthiocyanate on enzymes. PMID- 11946298 TI - Glycogen metabolism of isolated rat liver perfused with long-chain fatty acid. PMID- 11946299 TI - The oxidation of added NADH by intact heart mitochondria. PMID- 11946300 TI - Gel chromatography as a means for differentiation between dehydrogenases. PMID- 11946301 TI - Reduction kinetics and content of cytochrome P-450 by application of dual wavelength techniques to hemoglobin-free perfused rat liver. PMID- 11946303 TI - Collagen proline hydroxylation in chick embryos. PMID- 11946302 TI - Action of amobarbital on microsomal and mitochondrial respiratory state in perfused rat liver with and without phenobarbital induction. PMID- 11946304 TI - Incorporation of amino acids into mitochondrial protein of the flight muscle of Locusta migratoria in vitro and in vivo in the presence of cycloheximide. PMID- 11946305 TI - Crystallographic studies on luteoskyrine. PMID- 11946306 TI - Copper reduction by substrate in diamine oxidase. PMID- 11946307 TI - Kinetics of hydrolysis of uridylic oligonucleotides with different degree of polymerisation by snake venom phosphodiesterase. PMID- 11946308 TI - Resolution of 5'-mononucleotidase and non-specific phosphatase activities from skeletal muscles. PMID- 11946309 TI - Effect of insulin deficiency on the transport of glucose by rat small intestine. PMID- 11946310 TI - Some enzymatic properties of human brain acetylcholinesterase. PMID- 11946311 TI - Asynchronous development of different functions in mitochondria of saccharomyces cerevisiae. PMID- 11946313 TI - Inorganic pyrophosphate content in adult guinea pig liver. PMID- 11946312 TI - On the metabolic pathway of methylmethacrylate. PMID- 11946314 TI - Comparative aspects of fatty acid activation in Escherichia coli and Clostridium butyricum. PMID- 11946315 TI - Oscillations in the rate of hemoglobin synthesis in rabbit reticulocytes. PMID- 11946316 TI - Hydroxylation of poly(l-prolyl-l-prolylglycyl) of defined molecular weights by protocollagen proline hydroxylase. PMID- 11946317 TI - On the irreversible binding of p-(trimethylammonium) benzenediazonium fluoroborate (TDF) to acetylcholinesterase from electrogenic tissue. PMID- 11946318 TI - Isolation of two classes of rough vesicles from rat liver microsomes. PMID- 11946319 TI - ? AB - We have shown the existence in rat liver of free light cytoplasmic particles containing a stable, rapidly labelled RNA. These particles differ from the 45 S ribosomal fraction by several characters.The buoyant density and sedimentation properties suggest that these particles are similar to dRNA ribonucleoprotein particles released from polysomes and to those present in rat liver nuclei. PMID- 11946320 TI - ? AB - It is well known that, in rabbits, the allotypes of the b locus carried by the light chains of the IgG molecules cross-react. Nothing similar has been described for the allotypes of the a locus carried by the heavy chains of the IgG molecules. This paper describes a cross-reaction between the rabbit allotypic markers Aa1 and Aa2 of the a locus. This phenomenon became evident in successive I(125)-labelled IgG precipitation experiments with a particular serum. PMID- 11946321 TI - Heat denaturation of Bacillus Subtilis transforming DNA. PMID- 11946322 TI - The subunit size of alcohol dehydrogenates. PMID- 11946324 TI - A reinterpretation of the kinetics of pyruvate carboxylase. PMID- 11946325 TI - ? AB - Chromatography on carboxymethylcellulose of the trichloracetic extract "F(1)" of chicken erythrocytes nuclei, yields two histone fractions: F(1)-cF(1) the very lysine-rich histone and F(1)-cF(3), the fraction specific to erythrocytes.Aminoacid analysis and disc electrophorosis on extracts "F(1)" demonstrate that specific histone fraction F(1)-cF(3) occurs in smaller percentage in the early stages of chicken development; it increases from 6% (by weight of the total DNA) in 3 days old chicken to 16% in 21 days old chicken. But the percentage of F(1)-cF(1) fraction (8% by weight of total DNA) remains constant in the same conditions. PMID- 11946326 TI - Activation of a morphogenetic factor by electrophoresis. PMID- 11946323 TI - Molecular weight determination by chromatography on sepharose 4B. PMID- 11946327 TI - Minor constituents of human milk (I) identification of cyclohexaneundecanoic acid and phytanic acid in human milk fat by a combination gas chromatograph-mass spectrometer. PMID- 11946328 TI - Conversion of isocitrate to citrate and accumulation of the citrates in mitochondria. PMID- 11946329 TI - Polyribonucleotides containing a thiophosphate backbone. PMID- 11946331 TI - Properties of selenonicotinamide-adenine dinucleotide phosphate, an analogue of NADP. PMID- 11946330 TI - Formation of methyl esters by rat mucosa in methanol. PMID- 11946332 TI - The effect of a constitutive mutation in pseudomonas aeruginosa on rapidly labelled RNA. PMID- 11946333 TI - Isolation and properties of bromelin protease. PMID- 11946334 TI - Palmitoylcarnitine and tricarboxylic-acid-cycle oxidations. PMID- 11946335 TI - Stearate desaturation by microsomes on the locust fat-body. PMID- 11946336 TI - ? AB - The synthesis of fatty acids in mitochondria, microsomes and supernatant of mice liver was studied after administration of glucose 14C or (3)H, acetate 14C or 3H and pyruvate 14C.The results suggest that the cellular structures corresponding to the microsomes are the major site of fatty acid synthesis in the liver in vivo. PMID- 11946337 TI - Mechanisms in protein synthesis. VI: A method for measuring fast kinetics of binding of mRNA and aminoacyl-tRNA to ribosomes. PMID- 11946339 TI - The basic sequences of immunoglobulin kappa chains: Sequence studies of bence jones proteins Rad, Fr 4 and B6. PMID- 11946338 TI - Evidence for the presence of two forms of L-serine dehydratase in rat liver. PMID- 11946340 TI - The conformation of 4-thiouridine-5'-phosphate in single and double stranded polynucleotides. PMID- 11946341 TI - Chemical and mass spectrometric sequence studies of a peptide from the variable part of normal immunoglobulin lambda-chains. PMID- 11946342 TI - A "second messenger" for Vitamin D. PMID- 11946343 TI - The ATPase system of isolated rat liver plasma membranes. Kinetic properties of the MgATP phosphohydrolase. PMID- 11946344 TI - An immunochemical method for the characterization of polynucleotide phosphorylase from E. Coli. PMID- 11946345 TI - The effect of hydrocortisone on the accumulation of amylase in embryonic chick pancreas. PMID- 11946346 TI - The stimulation of the mitochondrial oxidation of a quinolphosphate by polylysine. PMID- 11946347 TI - Phylogenetic differences in the sensitivity of mitochondrial protein synthesising systems to antibiotics. PMID- 11946348 TI - Glucose-6-phosphate as regulator of monosaccharide transport in baker's yeast. PMID- 11946349 TI - C-methylation of desmethylmenaquinones: Specificity of the enzymatic system of mycobacterium phlei. PMID- 11946350 TI - Urate oxidation by cupric ion (Cu(++)). PMID- 11946351 TI - Pyruvate synthesis from acetyl coenzyme A and carbon dioxide with NADH(2) or NADPH(2) as electron donors. PMID- 11946352 TI - Stimulation of thymidylate kinase and deoxycytidylate deaminase activities of chang cells by deoxyadenosine. PMID- 11946353 TI - Inhibition of bacillus subtilis transforming system by acriflavin. PMID- 11946355 TI - Errata. PMID- 11946354 TI - The inhibition of chondroitin sulphate protein synthesis by cycloheximide. PMID- 11946356 TI - Foreword. PMID- 11946357 TI - Aims, applications and achievements. An introductory essay. PMID- 11946358 TI - Construction of biochemical computer models. AB - This paper described methods for constructing (digital) computer models of biochemical systems when the main object is to investigate the system itself, and not to fit experimental data (e.g., from tracer kinetics) to a set of equations. The author describes model-building as an art which is difficult to communicate, but nevertheless gives valuable tips on the conceptual and practical aspects, from his own considerable experience. PMID- 11946359 TI - Computer simulation of stochastic processes through model-sampling (Monte Carlo) techniques. AB - A simple Monte Carlo simulation program is outlined which can be used for the investigation of random-walk problems, for example in diffusion, or the movement of tracers in the blood circulation. The results given by the simulation are compared with those predicted by well-established theory, and it is shown how the model can be expanded to deal with drift, and with reflexion from or adsorption at a boundary. PMID- 11946360 TI - The numerical solution of stiff differential equations. AB - This paper first discusses the conditions in which a set of differential equations should give stable solutions, starting with linear systems assuming that these do not differ greatly in this respect from non-linear systems. Methods of investigating the stability of particular systems are briefly discussed. Most real biochemical systems are known from observation to be stable, but little is known of the regions over which stability persists; moreover, models of biochemical systems may not be stable, because of inaccurate choice of parameter values.The separate problem of stability and accuracy in numerical methods of approximating the solution of systems of non-linear equations is then treated. Stress is laid on the consistently unsatisfactory results given by explicit methods for systems containing "stiff" equations, and implicit multistep methods are particularly recommended for this class of problem, which is likely to include many biochemical model systems. Finally, an iteration procedure likely to give convergence both in multistep methods and in the steady-state approach is recommended, and areas in which improvement in methods is likely to occur are outlined. PMID- 11946361 TI - Steady states of general multi-enzyme networks and their associated properties. Computational approaches. AB - This paper discusses the theoretical basis for the determination of the steady state characteristics of a multi-enzyme network, given initial values for the enzyme and metabolite concentrations, and rate equations for each enzyme. The computation depends only to a small extent on integrating the rate equations, mainly on adjustment of the matrix of fluxes through each component of the system, until they are equal to each other within a prescribed tolerance limit.The program has been devised to be as flexible as possible to permit the user to investigate the effects of changing the concentration or kinetic properties of an individual enzyme. Alternatively, the concentrations of enzymes required to maintain a given flux may be computed. The program will deal with a network of up to about 30 enzymes. PMID- 11946362 TI - Observations on programs to estimate the parameters of enzyme kinetics. AB - This paper gives a critical account of the two major programs so far published specifically to estimate the parameters of enzyme kinetics. The dangers of submitting data to the programs without proper checks are discussed, and a screening test is described to identifty sets of data which may not be best fitted by a rectangular hyperbola. PMID- 11946363 TI - A survey of non-linear optimization techniques. AB - Optimization means the provision of a set of numerical parameter values which will give the best fit of an equation, or series of equations, to a set of data. For simple systems this can be done by differentiating the equations with respect to each parameter in turn, setting the set of partial differential equations to zero, and solving this set of simultaneous equations (as for exwnple in linear regression). In more complicated cases, however, it may be impossible to differentiate the equations, or very difficultly soluble non-linear equations may result. Many numerical optimization techniques to overcome these difficulties have been developed in the least ten years, and this review explains the logical basis of most of them, without going into the detail of computational procedures.The methods fall naturally into two classes - direct search methods, in which only values of the function to be minimized (or maximized) are used - and gradient methods, which also use derivatives of the function. The author considers all the accepted methods in each class, although warning that gradient methods should not be used unless the analytical differentiation of the function to be minimized is possible.If the solution is constrained, that is, certain values of the parameters are regarded as impossible or certain relations between the parameter values must be obeyed, the problem is more difficult. The second part of the review considers methods which have been proposed for the solution of constrained optimization problems. PMID- 11946364 TI - Kinetic modeling in physiology. AB - The author discusses the construction of model biochemical/physiological systems to fit experimental data which is always incomplete. He suggests that the first experiments on any system should be first-order (usually tracer) perturbations of steady-state systems. The proposed model can then be given a preliminary fit to the data; criteria for best fit, consistency, and uniqueness of fit are suggested. More data may then be obtained by perturbations at another steady state, and finally by studying transient situations.References are included to some of the author's own publications in which these matters are discussed in detail. PMID- 11946365 TI - An introduction to hybrid computers and their application to optimization problems. AB - The author, who is Secretary of the Analog Section of the British Computer Society, describes the way in which some of the logic components of a parallel logic analog computer function, and goes on to discuss the use of such a computer in optimization problems, as for instance in finding the best estimates of the parameters on compartmentation analysis of isotopic tracer experiments. The increased power which is gained when an analog computer is coupled to (hybridized with) a general purpose digital computer is then discussed, as shown for example in the increased speed of random-search optimization techniques when an analog computer is used to integrate the differential equations, or the improvement in steepest ascent methods when the parallel logic can be used to keep the hill climb path continuously on the line of steepest ascent. Finally, the use of a hybrid computer in solving partial differential equations is briefly outlined. PMID- 11946366 TI - Some experiences in the estimation of parameters in non-linear differential equations. AB - The author describes a procedure developed by himself and his colleagues for obtaining estimates of the parameters of rate equations, together with information about confidence regions for the estimates. The program has been used successfully for processing results from the chemical engineering industry, with highly non-linear model systems, particularly since temperature was a variable, and the "rate constants" were non-linear combinations of other constants. In biochemical situations, in which investigations are almost always at constant temperature, the non-linearity should not be so extreme, and the procedure may well be capable of dealing with more than 5 to 7 parameters for which it is recommended. PMID- 11946367 TI - Light dependent phosphorylation of rhodopsin by ATP. PMID- 11946368 TI - Calcium-dependent stimulation of prolactin release in rat anterior pituitary in vitro by n(6)-monobutyryl adenosine 3',5'-monophosphate. PMID- 11946370 TI - Mutants of Aspergillus nidulans lacking pyruvate carboxylase. PMID- 11946371 TI - Some effects of glucocorticoids on the subcellular distribution of the activities of citrate synthase and phosphoenolpyruvate carboxykinase in livers of rats and cows. PMID- 11946372 TI - Interaction between concanavalin A and heparin. PMID- 11946373 TI - Isolation of lipoprotein A with hydrated density characteristic for low density lipoproteins. PMID- 11946374 TI - In vitro effect of NAD on DNa synthesis in isolated nuclei from regenerating rat liver and novikoff hepatoma. PMID- 11946375 TI - Comparison of aminoacyl-tRNA synthetases from rat liver and hepatomas. PMID- 11946376 TI - Binding-catalysis relationship in alpha-chymotrypsin action as revealed from reversible inhibition study of phenylalkylboronic acids. PMID- 11946377 TI - A reexamination of the reaction of TPCK with alpha-chymotrypsin. PMID- 11946378 TI - A beta-mannosidic linkage in the carbohydrate moiety of ovalbumin. PMID- 11946379 TI - Characteristics of DNA-dependent RNA polymerase activity from isolated yeast nuclei. PMID- 11946380 TI - Biliverdin photo-oxidation. In vitro formation of methylvinylmaleimide. PMID- 11946381 TI - DNA and RNA from avian myeloblastosis virus as templates for viral DNA polymerase. PMID- 11946382 TI - An energy-dependent transformation of a ferricytochrome of the mitochondrial respiratory chain. PMID- 11946383 TI - Exposed and buried guanosine residues in tRNA(1)(Val) from yeast. PMID- 11946384 TI - Proteins of human glial cell membrane. PMID- 11946385 TI - Conversion of squalene into sterols by microsomal fractions from brains of developing rats. PMID- 11946386 TI - Plasmalogen biosynthesis in a cell-free system. Enzymic desaturation of 1-O-alkyl (2-acyl) glycerophosphoryl ethanolamine. PMID- 11946388 TI - Isolation and partial characterization of several different chorionic gonadotropin (HCG) components. PMID- 11946387 TI - Lipolytic activity of bovine growth hormone bound to sepharose beads. PMID- 11946389 TI - Evidence for pentacoordinated iron (II) in carboxymethylated cytochrome c. PMID- 11946390 TI - Monomer and magnetic dipole-coupled Cu(2+) EPR signals in nitrosylhemocyanin. PMID- 11946391 TI - Incorporation of plant sterols into membranes and its relation to sterol absorption. PMID- 11946393 TI - Effect of triton X-100 and trypsin on NADPH-cytochrome C reductase reconstitutively active in fatty acid omega-hydroxylation. PMID- 11946392 TI - Immunological evidences for a localization of system I on the outside face and of system II on the inside face of the chloroplast lamella. PMID- 11946394 TI - "Cytoplasmic" DNA from primary embryonic cell cultures is not informational. PMID- 11946395 TI - C-phycocyanin from the thermophilic blue-green alga Mastigocladus laminosus, isolation, characterization and subunit composition. PMID- 11946397 TI - Errata. PMID- 11946396 TI - A simple method for the large-scale preparation of sucrose gradients. PMID- 11946398 TI - Teaching of biochemistry and education of biochemists. PMID- 11946399 TI - A Dicarboxyclic acid transport system in Bacillus subtilis. PMID- 11946400 TI - Protein kinases from rat skeletal muscle: Evidence for six different fractions of the enzyme. PMID- 11946401 TI - Nucleoside conformations VI. Optical properties of 1,3-diribofuranosyl-6-methyl uracil. PMID- 11946402 TI - Effect of alpha-amanitin on chromosomal and nucleolar RNA-synthesis in Chironomus thummi polytene chromosomes. PMID- 11946403 TI - Purification of a soluble ATPase of rat liver. PMID- 11946404 TI - Biosynthesis of phosphatidylglycerol during greening of synchronous dark-grown Euglena gracilis Z. PMID- 11946406 TI - Phosphohistidine as the result of phosphate migration in phosphorylated inorganic pyrophosphatase from yeast. PMID- 11946405 TI - Biosynthesis of actinomycin D : Purification and properties of an enzyme which activates L-valine. PMID- 11946407 TI - Enzymic synthesis of 9- and 7-(2'-beta-D-deoxyribosyl) xanthine. PMID- 11946408 TI - Pullulan elaboration, an inducible system of Pullularia pullulans. PMID- 11946410 TI - Possible function of the protein bound to nuclear complementary RNA. PMID- 11946409 TI - Heterogeneity of chromosomal RNA. PMID- 11946412 TI - Influence of enzyme concentration on the reaction of rabbit muscle phosphofructokinase with antibodies. PMID- 11946411 TI - Activation of rabbit muscle phosphofructokinase by specific antibodies. PMID- 11946413 TI - Insulin action on the adenyl cyclase system: Antagonism to activation by lipolytic hormones. PMID- 11946414 TI - The utilization of intramitochondrially generated carbamyl phosphate for microsomal glucose 6-phosphate biosynthesis. PMID- 11946415 TI - Molecular weight estimations of proteins by electrophoresis in polyacrylamide gels of graded porosity. PMID- 11946416 TI - Primary incorporation of U-(14)C-glucose into glucose-1-phosphate and glucose-6 phosphate by incubated pigeon liver homogenate. PMID- 11946417 TI - Demonstration of an "activator factor" and an "inhibitor factor" in the cyclic AMP phosphodiesterase from oxyntic cells of bullfrog gastric mucosa. PMID- 11946418 TI - Association of ribosomal RNA with ribosomal proteins covalently bound to sepharose. PMID- 11946419 TI - The photoinhibition of malate dehydrogenase. PMID- 11946420 TI - Photooxidation of the thioether bridges of horse heart cytochrome C. PMID- 11946421 TI - Prevention of ribosomal donor site from occupation by aminoacyl tRNA during polypeptide synthesis. PMID- 11946422 TI - The kinetics of potassium ion complexation by ionophores. PMID- 11946423 TI - Effects of L-leucine and alpha-ketoisocaproic acid upon insulin secretion and metabolism of isolated pancreatic islets. PMID- 11946424 TI - On the inhibition of mitochondrial electron transport by Zn(2+) ions.